PMID- 15876201 TI - Uncertainty and sensitivity analysis in assessment of the thyroid cancer risk related to Chernobyl fallout in Eastern France. AB - The increase in the thyroid cancer incidence in France observed over the last 20 years has raised public concern about its association with the 1986 nuclear power plant accident at Chernobyl. At the request of French authorities, a first study sought to quantify the possible risk of thyroid cancer associated with the Chernobyl fallout in France. This study suffered from two limitations. The first involved the lack of knowledge of spontaneous thyroid cancer incidence rates (in the absence of exposure), which was especially necessary to take their trends into account for projections over time; the second was the failure to consider the uncertainties. The aim of this article is to enhance the initial thyroid cancer risk assessment for the period 1991-2007 in the area of France most exposed to the fallout (i.e., eastern France) and thereby mitigate these limitations. We consider the changes over time in the incidence of spontaneous thyroid cancer and conduct both uncertainty and sensitivity analyses. The number of spontaneous thyroid cancers was estimated from French cancer registries on the basis of two scenarios: one with a constant incidence, the other using the trend observed. Thyroid doses were estimated from all available data about contamination in France from Chernobyl fallout. Results from a 1995 pooled analysis published by Ron et al. were used to determine the dose-response relation. Depending on the scenario, the number of spontaneous thyroid cancer cases ranges from 894 (90% CI: 869-920) to 1,716 (90% CI: 1,691-1,741). The number of excess thyroid cancer cases predicted ranges from 5 (90% UI: 1-15) to 63 (90% UI: 12-180). All of the assumptions underlying the thyroid cancer risk assessment are discussed. PMID- 15876202 TI - Residential radon in Canada: an uncertainty analysis of population and individual lung cancer risk. AB - Following a comprehensive evaluation of the health risks of radon, the U.S. National Research Council (US-NRC) concluded that the radon inside the homes of U.S. residents is an important cause of lung cancer. To assess lung cancer risks associated with radon exposure in Canadian homes, we apply the new (US-NRC) techniques, tailoring assumptions to the Canadian context. A two-dimensional uncertainty analysis is used to provide both population-based (population attributable risk, PAR; excess lifetime risk ratio, ELRR; and life-years lost, LYL) and individual-based (ELRR and LYL) estimates. Our primary results obtained for the Canadian population reveal mean estimates for ELRR, PAR, and LYL are 0.08, 8%, and 0.10 years, respectively. Results are also available and stratified by smoking status (ever versus never). Conveniently, the three indices (ELRR, PAR, and LYL) reveal similar output uncertainty (geometric standard deviation, GSD approximately 1.3), and in the case of ELRR and LYL, comparable variability and uncertainty combined (GSD approximately 4.2). Simplifying relationships are identified between ELRR, LYL, PAR, and the age-specific excess rate ratio (ERR), which suggest a way to scale results from one population to another. This insight is applied in scaling our baseline results to obtain gender-specific estimates, as well as in simplifying and illuminating sensitivity analysis. PMID- 15876203 TI - Using physiologically-based pharmacokinetic modeling to address nonlinear kinetics and changes in rodent physiology and metabolism due to aging and adaptation in deriving reference values for propylene glycol methyl ether and propylene glycol methyl ether acetate. AB - Reference values, including an oral reference dose (RfD) and an inhalation reference concentration (RfC), were derived for propylene glycol methyl ether (PGME), and an oral RfD was derived for its acetate (PGMEA). These values were based on transient sedation observed in F344 rats and B6C3F1 mice during a two year inhalation study. The dose-response relationship for sedation was characterized using internal dose measures as predicted by a physiologically based pharmacokinetic (PBPK) model for PGME and its acetate. PBPK modeling was used to account for changes in rodent physiology and metabolism due to aging and adaptation, based on data collected during Weeks 1, 2, 26, 52, and 78 of a chronic inhalation study. The peak concentration of PGME in richly perfused tissues (i.e., brain) was selected as the most appropriate internal dose measure based on a consideration of the mode of action for sedation and similarities in tissue partitioning between brain and other richly perfused tissues. Internal doses (peak tissue concentrations of PGME) were designated as either no-observed adverse-effect levels (NOAELs) or lowest-observed-adverse-effect levels (LOAELs) based on the presence or the absence of sedation at each time point, species, and sex in the two-year study. Distributions of the NOAEL and LOAEL values expressed in terms of internal dose were characterized using an arithmetic mean and standard deviation, with the mean internal NOAEL serving as the basis for the reference values, which was then divided by appropriate uncertainty factors. Where data were permitting, chemical-specific adjustment factors were derived to replace default uncertainty factor values of 10. Nonlinear kinetics, which was predicted by the model in all species at PGME concentrations exceeding 100 ppm, complicate interspecies, and low-dose extrapolations. To address this complication, reference values were derived using two approaches that differ with respect to the order in which these extrapolations were performed: (1) default approach of interspecies extrapolation to determine the human equivalent concentration (PBPK modeling) followed by uncertainty factor application, and (2) uncertainty factor application followed by interspecies extrapolation (PBPK modeling). The resulting reference values for these two approaches are substantially different, with values from the latter approach being seven-fold higher than those from the former approach. Such a striking difference between the two approaches reveals an underlying issue that has received little attention in the literature regarding the application of uncertainty factors and interspecies extrapolations to compounds where saturable kinetics occur in the range of the NOAEL. Until such discussions have taken place, reference values based on the former approach are recommended for risk assessments involving human exposures to PGME and PGMEA. PMID- 15876204 TI - Quantitative estimates of risk for noncancer endpoints. AB - While quantitative estimates of risk have been a standard practice in cancer risk assessment for many years, no similar practice is evident in noncancer risk assessment. We use two recent examples involving methylmercury and arsenic to illustrate the negative impact of this discrepancy on risk communication and cost benefit analysis. We argue for a more balanced treatment of cancer and noncancer risks and suggest an approach for reaching this goal. PMID- 15876205 TI - Model uncertainty and risk estimation for experimental studies of quantal responses. AB - Experimental animal studies often serve as the basis for predicting risk of adverse responses in humans exposed to occupational hazards. A statistical model is applied to exposure-response data and this fitted model may be used to obtain estimates of the exposure associated with a specified level of adverse response. Unfortunately, a number of different statistical models are candidates for fitting the data and may result in wide ranging estimates of risk. Bayesian model averaging (BMA) offers a strategy for addressing uncertainty in the selection of statistical models when generating risk estimates. This strategy is illustrated with two examples: applying the multistage model to cancer responses and a second example where different quantal models are fit to kidney lesion data. BMA provides excess risk estimates or benchmark dose estimates that reflects model uncertainty. PMID- 15876206 TI - Development of the PEARLS model (Particulate Exposure from Ambient to Regional Lung by Subgroup) and use of Monte Carlo simulation to predict internal exposure to 2.5 in Toronto. AB - Air pollution is a current and growing concern for Canadians, and there is evidence that ambient levels that meet current exposure standards may be associated with mortality and morbidity in Toronto, Canada. Evaluating exposure is an important step in understanding the relationship between particulate matter (PM) exposure and health outcomes. This report describes the PEARLS model (Particulate Exposure from Ambient to Regional Lung by Subgroup), which predicts exposure distributions for 11 age-gender population subgroups in Toronto to PM2.5 (PM with a median aerodynamic diameter of 2.5 microm or less) using Monte Carlo simulation techniques. The model uses physiological and activity pattern characteristics of each subgroup to determine region-specific lung exposure to PM2.5, which is defined as the mass of PM2.5 deposited per unit time to each of five lung regions (two extrathoracic, bronchial, bronchiolar, and alveolar). The modeling results predict that children, toddlers, and infants have the broadest distributions of exposure, and the greatest chance of experiencing extreme exposures in the alveolar region of the lung. Importance analysis indicates that the most influential model variables are air exchange rate into indoor environments, time spent outdoors, and time spent at high activity levels. Additionally, a "critical point" was defined and introduced to the PEARLS to investigate the effects of possible threshold-pathogenic phenomena on subgroup exposure patterns. The analysis indicates that the subgroups initially predicted to be most highly exposed were likely to have the highest proportion of their population exposed above the critical point. Substantial exposures above the critical point were predicted in all subgroups for ambient concentrations of PM2.5 commonly observed in Toronto after continuous exposure of 24 hours or more. PMID- 15876207 TI - Probability distributions for showering and bathing water-use behavior for various U.S. subpopulations. AB - It has been shown that bathroom-type water uses dominate personal exposure to water-borne contaminants in the home. Therefore, in assessing exposure of specific population groups to the contaminants in the water, understanding population water-use behavior for bathroom activities as a function of demographic characteristics is vital to realistic exposure estimates. In this article, shower and bath frequencies and durations are analyzed, presented, and compared for various demographic groups derived from analyses of the National Human Activities Pattern Survey (NHAPS) database and the Residential End Uses of Water Study (REUWS) database as well as from a review of current literature. Analysis showed that age and level of education significantly influenced shower and bath frequency and duration. The frequency of showering and bathing reported in NHAPS agreed reasonably well with previous studies; however, durations of these events were found to be significantly longer. Showering frequency reported in REUWS was slightly less than that reported for NHAPS; however, durations of showers reported in REUWS are consistent with other studies. After considering the strengths and weaknesses of each data set and comparing their results to previous studies, it is concluded that NHAPS provides more reliable frequency data, while REUWS provides more reliable duration data. The shower- and bath-use behavior parameters recommended in this article can aid modelers in appropriately specifying water-use behavior as a function of demographic group in order to conduct reasonable assessments of exposure to contaminants that enter the home via the water supply. PMID- 15876208 TI - A multilayered box model for calculating preliminary remediation goals in soil screening. AB - In the process of screening a soil against a certain contaminant, we define the health-risk-based preliminary remediation goal (PRG) as the contaminant concentration above which some remedial action may be required. Thus, PRG is the first standard (or guidance) for judging a site. An overestimated PRG (a too large value) may cause us to miss some contaminated sites that can threaten human health and the environment. An underestimated PRG (a too-small value), on the other hand, may lead to unnecessary cleanup and waste tremendous resources. The PRGs for soils are often calculated on the assumption that the contaminant concentration in soil does not change with time. However, that concentration usually decreases with time as a result of different chemical and transport mechanisms. The static assumption thus exaggerates the long-term exposure dose and results in a too-small PRG. We present a box model that considers all important transport processes and obeys the law of mass conservation. We can use the model as a tool to estimate the transient contaminant concentrations in air, soil, and ground water. Using these concentrations in conjunction with appropriate health-risk parameters, we may estimate the PRGs for different contaminants. As an example, we calculated the tritium PRG for residential soils. The result is quite different from, but within the range of, the two versions of the corresponding PRG previously recommended by the U.S. EPA. PMID- 15876209 TI - Additivity of information value in two-act linear loss decisions with normal priors. AB - Information about two uncertainties is superadditive in value if the value of information for resolving both uncertainties together exceeds the sum of the value of information for resolving each uncertainty alone. For the two-act linear loss decision problem with normal priors, conditions are derived for which the expected value of perfect information about two independent risks is superadditive. An approximate condition is also presented. The level of additivity of information value has implications for how risk-management resources should be allocated to acquisition of information. Several applications show how a variety of decision problems can reduce to the basic problem, and how the general results obtained here can be translated simply to prescriptions for specific situations. PMID- 15876210 TI - A screening methodology for the identification and ranking of infrastructure vulnerabilities due to terrorism. AB - The extreme importance of critical infrastructures to modern society is widely recognized. These infrastructures are complex and interdependent. Protecting the critical infrastructures from terrorism presents an enormous challenge. Recognizing that society cannot afford the costs associated with absolute protection, it is necessary to identify and prioritize the vulnerabilities in these infrastructures. This article presents a methodology for the identification and prioritization of vulnerabilities in infrastructures. We model the infrastructures as interconnected digraphs and employ graph theory to identify the candidate vulnerable scenarios. These scenarios are screened for the susceptibility of their elements to a terrorist attack, and a prioritized list of vulnerabilities is produced. The prioritization methodology is based on multiattribute utility theory. The impact of losing infrastructure services is evaluated using a value tree that reflects the perceptions and values of the decisionmaker and the relevant stakeholders. These results, which are conditional on a specified threat, are provided to the decisionmaker for use in risk management. The methodology is illustrated through the presentation of a portion of the analysis conducted on the campus of the Massachusetts Institute of Technology. PMID- 15876211 TI - Compressed natural gas bus safety: a quantitative risk assessment. AB - This study assesses the fire safety risks associated with compressed natural gas (CNG) vehicle systems, comprising primarily a typical school bus and supporting fuel infrastructure. The study determines the sensitivity of the results to variations in component failure rates and consequences of fire events. The components and subsystems that contribute most to fire safety risk are determined. Finally, the results are compared to fire risks of the present generation of diesel-fueled school buses. Direct computation of the safety risks associated with diesel-powered vehicles is possible because these are mature technologies for which historical performance data are available. Because of limited experience, fatal accident data for CNG bus fleets are minimal. Therefore, this study uses the probabilistic risk assessment (PRA) approach to model and predict fire safety risk of CNG buses. Generic failure data, engineering judgments, and assumptions are used in this study. This study predicts the mean fire fatality risk for typical CNG buses as approximately 0.23 fatalities per 100-million miles for all people involved, including bus passengers. The study estimates mean values of 0.16 fatalities per 100-million miles for bus passengers only. Based on historical data, diesel school bus mean fire fatality risk is 0.091 and 0.0007 per 100-million miles for all people and bus passengers, respectively. One can therefore conclude that CNG buses are more prone to fire fatality risk by 2.5 times that of diesel buses, with the bus passengers being more at risk by over two orders of magnitude. The study estimates a mean fire risk frequency of 2.2 x 10(-5) fatalities/bus per year. The 5% and 95% uncertainty bounds are 9.1 x 10(-6) and 4.0 x 10(-5), respectively. The risk result was found to be affected most by failure rates of pressure relief valves, CNG cylinders, and fuel piping. PMID- 15876212 TI - Risk analysis of Finnish peacekeeping in Kosovo. AB - The research team interviewed over 90 Finnish battalion members in Kosovo, visited 22 units or posts, registered its observations, and made any necessary measurements. Key persons were asked to list the most important risks for occupational safety and health in their area of responsibility. Altogether, 106 accidents and 40 cases of disease resulted in compensation claims in 2000. The risks to the peacekeeping force were about twice those of the permanent staff of military trainees in Finland. Altogether, 21 accidents or cases of disease resulted in sick leave for at least 3 months after service. One permanent injury resulted from an explosion. Biological, chemical, and physical factors caused 8 to 9 occupational illnesses each. Traffic accidents, operational factors, and munitions and mines were evaluated to be the three most important risk factors, followed by occupational hygiene, living conditions (mold, fungi, dust), and general hygiene. Possible fatal risks, such as traffic accidents and munitions and explosives, received a high ranking in both the subjective and the objective evaluations. One permanent injury resulted from an explosion, and two traffic accidents involved a fatality, although not of a peacekeeper. The reduction of sports and military training accidents, risk-control programs, and, for some tasks, better personal protection is considered a development challenge for the near future. PMID- 15876213 TI - Predictive genetic testing for Alzheimer's disease: impact upon risk perception. AB - The aim of this study was to determine the impact on risk perceptions of disclosing genetic test results used to estimate the risk of Alzheimer's disease (AD). Adult children (n = 149) of people with AD were randomized to one of two groups--Intervention group: lifetime risk estimates of AD based on age, gender, family history, and Apolipoprotein E (APOE) genotype; CONTROL GROUP: lifetime risk estimates of AD based on the same risk factors excluding APOE genotype. Perceptions of personal risk (PPR) for AD were assessed six weeks after risk assessments. PPR were correlated with actual lifetime risk estimates (r = 0.501; p < 0.0001). After controlling for lifetime risks communicated to participants, age, and number of affected relatives, PPR scores among those with an epsilon4 positive test result (the test result associated with increased AD susceptibility) (adjusted mean: 3.4 (SD: 0.7)) were not different from the PPR scores in the CONTROL GROUP (adjusted mean: 3.4 (SD: 0.7) (F1,91= 1.98; p = 0.162). Again, controlling for lifetime risk estimates, age, and number of affected relatives, the PPR score of those receiving an epsilon4-negative test result was significantly lower (adjusted mean: 3.1 (SD: 0.8)) than those in the CONTROL GROUP (adjusted mean: 3.4 (SD: 0.7) (F1,95 = 6.23; p = 0.014). Perceptions of risk of developing AD are influenced by genetic test disclosure in those receiving epsilon4-negative, but not those receiving epsilon4-positive test results. Despite the reduced perceptions of risk in the former group, there was no evidence of false reassurance (i.e., perceiving risks as equal to or lower than population risks of AD), although this possibility should be assessed in other testing contexts. PMID- 15876214 TI - Aggregate, disaggregate, and hybrid analyses of ecological risk perceptions. AB - Laypeople's perceptions of health and safety risks have been widely studied, but only a few studies have addressed perceptions of ecological hazards. We assembled a list of 39 attributes of ecological hazards from the literatures on comparative risk assessment, ecological health, environmental conservation and management, environmental psychology, and risk perception. In Study 1, 125 laypeople evaluated 83 hazards on subsets of this attribute set. Factor analysis of attribute ratings (averaged over participants) revealed six oblique factors: ecological impacts, human impacts, human benefits, aesthetic impacts, scientific understanding, and controllability. These factors predicted mean judgments of overall riskiness, ecological riskiness, acceptability, and regulatory strictness. In Study 2, 30 laypeople each evaluated 34 hazards on 17 attributes and 3 dependent variables. Aggregate-level factor analysis of these data replicated the appropriate portion of the factor solution and yielded similar regression results. Parallel analyses at the individual-participant level yielded factors that explained less variance in judgments of overall riskiness, ecological riskiness, and acceptability. However, the decrease in explanatory power was much less than is often reported for disaggregate-level analyses of psychometric data. This discrepancy illustrates the importance of distinguishing between the level of analysis (aggregate versus disaggregate) and the focus of analysis (distinctions among hazards versus distinctions among participants). In a hybrid analysis, aggregate-level factor scores predicted individual participants' riskiness judgments reasonably well. Psychometric studies such as these provide a sound empirical basis for selecting attributes of ecological hazards for use in comparative risk assessment. PMID- 15876215 TI - Using integrated geospatial mapping and conceptual site models to guide risk based environmental clean-up decisions. AB - Government and private sector organizations are increasingly turning to the use of maps and other visual models to provide a depiction of environmental hazards and the potential risks they represent to humans and ecosystems. Frequently, the graphic presentation is tailored to address a specific contaminant, its location and possible exposure pathways, and potential receptors. Its format is usually driven by the data available, choice of graphics technology, and the audience being served. A format that is effective for displaying one contaminant at one scale at one site, however, may be ineffective in accurately portraying the circumstances surrounding a different contaminant at the same site, or the same contaminant at a different site, because of limitations in available data or the graphics technology being used. This is the daunting challenge facing the U.S. Department of Energy (DOE), which is responsible for the nation's legacy wastes from nuclear weapons research, testing, and production at over 100 sites in the United States. In this article, we discuss the development and use of integrated geospatial mapping and conceptual site models to identify hazards and evaluate alternative long-term environmental clean-up strategies at DOE sites located across the United States. While the DOE probably has the greatest need for such information, the Department of Defense and other public and private responsible parties for many large and controversial National Priority List or Superfund sites would benefit from a similar approach. PMID- 15876216 TI - Toward cost-benefit analysis of acute behavioral effects of toluene in humans. AB - There is increasing interest in being able to express the consequences of exposure to potentially toxic compounds in monetary terms in order to evaluate potential cost-benefit relationships of controlling exposure. Behavioral effects of acute toluene exposure could be subjected to cost-benefit analysis if the effects of toluene were quantitatively compared to those of ethanol ingestion, which has been monetized for applied contexts. Behavioral effects of toluene and ethanol were quantified by meta-analysis of studies from the peer-reviewed literature describing their effects on choice reaction time (reaction time in a test requiring a subject to choose among two or more alternatives before responding). The internal doses of these compounds were estimated by a general physiological and toxicokinetic (GPAT) simulation from exposure parameters provided in the reports. The reported effects were converted to a common metric (proportion of baseline) and related to the estimated internal doses of toluene and ethanol, from which dose-effect equations were fitted. The estimated effect of toluene was compared to the estimated effect of ethanol on the same dependent variable by deriving a dose-equivalence equation (DEE) to express the dose of toluene as an equivalent dose of ethanol on the basis of equal effect magnitude. A nomogram was constructed by GPAT simulation to relate the environmental exposure concentration of toluene to the equivalent effect magnitude of a range of ethanol internal doses. Behavioral effects and their evaluation are determined by internal doses, which in turn are determined by a variety of variables. In addition to concentration and duration of exposure, which determine internal dose by pharmacokinetic processes, the activity level of exposed persons is a major factor. This analysis provides a continuous function of the consequences of toluene exposure expressed as ethanol-equivalent doses within confidence limits. The resulting function has the potential to estimate the monetary values of behavioral deficits caused by a range of exposures to toluene from existing monetized information on ethanol. PMID- 15876217 TI - Bringing social standards into project evaluation under dynamic uncertainty. AB - Society often sets social standards that define thresholds of damage to society or the environment above which compensation must be paid to the state or other parties. In this article, we analyze the interdependence between the use of social standards and investment evaluation under dynamic uncertainty where a negative externality above a threshold established by society requires an assessment and payment of damages. Under uncertainty, the party considering implementing a project or new technology must not only assess when the project is economically efficient to implement but when to abandon a project that could potentially exceed the social standard. Using real-option theory and simple models, we demonstrate how such a social standard can be integrated into cost benefit analysis through the use of a development option and a liability option coupled with a damage function. Uncertainty, in fact, implies that both parties interpret the social standard as a target for safety rather than an inflexible barrier that cannot be overcome. The larger is the uncertainty, in fact, the greater will be the tolerance for damages in excess of the social standard from both parties. PMID- 15876218 TI - Using surveys in public participation processes for risk decision making: the case of the 2003 British GM Nation? Public debate. AB - This article takes as its case study the "GM Nation?" public debate, a major participation process on the commercialization of agricultural biotechnology, which occurred in Britain during the summer of 2003. We investigate possible self selection biases in over 36,000 open questionnaire responses on the risks and benefits of genetically modified crops and food obtained during GM Nation? A comparison sample of equivalent responses from a statistically representative sample (n = 1,363) of the British general public obtained shortly after the conclusion of the debate is reported. This comparison shows that the GM Nation? open responses were indeed not fully representative of British "public opinion" regarding agricultural biotechnology. Rather, such opinion is not a unitary whole, but fragmented, with considerable ambivalence coexisting alongside outright opposition to GM agriculture. The methodological implications for multistage participation processes are discussed: in particular, the need to anticipate outcomes of complex design decisions, and to include representative public surveys as standard where measures of broader public attitudes to risk are an important objective. PMID- 15876219 TI - Combining quantitative and qualitative measures of uncertainty in model-based environmental assessment: the NUSAP system. AB - This article discusses recent experiences with the Numeral Unit Spread Assessment Pedigree (NUSAP) system for multidimensional uncertainty assessment, based on four case studies that vary in complexity. We show that the NUSAP method is applicable not only to relatively simple calculation schemes but also to complex models in a meaningful way and that NUSAP is useful to assess not only parameter uncertainty but also (model) assumptions. A diagnostic diagram can be used to synthesize results of quantitative analysis of parameter sensitivity and qualitative review (pedigree analysis) of parameter strength. It provides an analytic tool to prioritize uncertainties according to quantitative and qualitative insights in the limitations of available knowledge. We show that extension of the pedigree scheme to include societal dimensions of uncertainty, such as problem framing and value-laden assumptions, further promotes reflexivity and collective learning. When used in a deliberative setting, NUSAP pedigree assessment has the potential to foster a deeper social debate and a negotiated management of complex environmental problems. PMID- 15876221 TI - Use of monoclonal antibodies in blocking ELISA detection of transmissible gastroenteritis virus in faeces of piglets. AB - Monoclonal antibodies (mAb) to the transmissible gastroenteritis virus (TGEV) nucleoprotein (N) and membrane protein (M) were prepared and used for the comparative assessment of three blocking ELISA variants to detect TGEV. The competitive blocking ELISA format showed the highest sensitivity, allowing detection of 10(3) TCID50 TGEV/ml in culture medium. Ninety-nine porcine field faecal samples obtained from 37 herds affected with diarrhoea were examined, and various TGEV levels were found in nine samples from six herds. However, only in three samples were significant TGEV concentrations demonstrated. The relationship between incidence of TGEV gastroenteritis and the spread of porcine respiratory coronavirus infection in pig farms is discussed. PMID- 15876222 TI - A novel subgroup among genotypes of equine arteritis virus: genetic comparison of 40 strains. AB - The authors determined partial nucleic sequences of the variable regions of open reading frame (ORF5) from 151 nucleotide to 668 nucleotide and deduced amino acid sequences of 518 nucleotide respectively of 20 equine arteritis virus (EAV) isolates. About 19 Hungarian and one Austrian EAV strains were subjected to sequence analysis, the further data of 20 EAV strains: six North American and 14 European were obtained from the GenBank. Comparative sequence analysis of the Hungarian EAV strains indicated that among the three variable regions the first has been affected mostly by point mutations. Genetic comparison of the Hungarian strains with other EAV isolates from western Europe and North America (including the Bucyrus reference strain) has been performed on the aforementioned genome region. Besides the already known genetic subgroups of EAV; phylogenetic analysis revealed a novel subgroup comprising mainly Hungarian strains. Compared with the Bucyrus virus, the overall sequence divergencies of the examined Hungarian strains ranged from 81.47 to 90.73% at nucleotide and from 84.88 to 91.86% at amino acid level. Epizootiological studies have shown that the significant part of the EAV strains having been existed in Hungary before and in 2000 belong to this unique cluster (II.D) which was not indicated in former phylogenetic studies. After 2000 new EAV strains emerged in Hungary, one of them causing abortions or neonatal death. The previously dominant 'Hungarian' EAV genotypes were replaced by these new strains belonging to North American and European subgroups (I.A, I.B, II.A, II.B). The anamnesis of these cases revealed connections with persistent virus shedder stallions, those were imported to the country after 2000 or have been infected abroad. One of these Hungarian stallions became the source of abortion storms in Hungarian studs. PMID- 15876223 TI - Investigations into the basis of chloramphenicol and tetracycline resistance in Staphylococcus intermedius isolates from cases of pyoderma in dogs. AB - A total of 160 Staphylococcus intermedius isolates were recovered from cases of pyoderma in 2002 and were examined for susceptibility to 13 different antimicrobial agents. Ninety per cent (144) of the isolates were resistant to tetracycline, derivatives of which have been used until recently, and 18% (29) were resistant to chloramphenicol which was banned from use 13 years ago. The presence of genes encoding chloramphenicol acetyltransferase (CAT) and tetracycline resistance (tet); tet(K), (L), (M), and (O) were determined by PCR in the 29 chloramphenicol and tetracycline resistant isolates. Seventeen (59%) isolates contained the cat gene while 12 (41%) isolates did not carry the cat gene, implying there may be other genes for chloramphenicol resistance that were not detected by the primers (primer set 1) used in this study. The tet(M) gene was found in 28 (97%) of the resistant S. intermedius isolates, but none contained the tet(O) gene. All 29 isolates carried one or two tet genes; tet(K), (L), and (M), with four different distribution patterns. New PCR products, a 1.1 kb product using primer set 1 and a 0.2 kb product using primer set 2, were cloned and sequenced. A 904 bp fragment of S. aureus plamid pS194, including sequence from the streptomycin adenyltransferase gene (804 bp), was found inserted into the terminal region of the cat gene (GenBank accession no. AY604739), whilst the sequence of 0.2 kb was previously unpublished. PMID- 15876224 TI - Anti-microbial susceptibility of Streptococcus spp. isolated from bovine mastitis in Argentina. AB - The in vitro susceptibility to penicillin G, erythromycin and clindamycin was determined by the disc diffusion test and by E-test for a total of 47 streptococcal strains (three Streptococcus uberis, 36 Streptococcus agalactiae, eight Streptococcus dysgalactiae spp. dysgalactiae) isolated from bovine intramammary infections in Argentina. Moreover, resistance phenotypes of erythromycin-resistant streptococcal isolates was characterized. MIC90 of penicillin G, erythromycin and clindamycin for S. agalactiae were 0.75, 8.0 and 12.0 microg/ml respectively. Resistance to erythromycin and clindamycin was detected in 13 (27.6%) and 12 (25.5%) isolates respectively. No isolate was resistant to penicillin G. Resistance against macrolides, lincosamides and streptogramin B (MLS(B)) represented by the constitutive MLS(B) phenotype was present in 11 (23.4%) erythromycin-resistant isolates and two isolates (4.3%) expressed the M phenotype. The inducible MLS(B) phenotype was not identified. Results suggest that beta-lactams are the first-line antibiotics when treating streptococcal udder infections; however, the continuous monitoring of the antibiotic resistance is essential, as the emergence of resistant strains has become a growing concern on the therapy of bovine mastitis. PMID- 15876225 TI - Molecular typing of Mycobacterium bovis isolates from south-east Brazil by spoligotyping and RFLP. AB - The identification of 163 strains of Mycobacterium bovis by polymerase chain reaction (PCR) and microbiological tests was carried out on 252 tuberculous-like lesions (TLLs) collected from slaughtered cattle in south-east Brazil. This study compared the usefulness of three genotyping techniques, IS6110-restriction fragment length polymorphism (RFLP), polymorphic guanine-cytosine-rich sequence (PGRS)-RFLP and direct repeat (DR)-spoligotyping, as applied to M. bovis isolates. Based on IS6110-RFLP genotyping we selected a group of 23 isolates containing more than one IS6110 copy, along with 16 samples containing one IS6110 copy from different geographical areas, evenly distributed among dairy (eight) and beef cattle (eight). These selected isolates were analysed by PGRS-RFLP and DR-spoligotyping genotyping. Dairy cattle (17%) display a higher frequency of multiple IS6110 copies than beef cattle (10%). A comparison between the genotype data obtained fails to show a correlation between the main clusters found by the three techniques. However, the clustering of each genotyping procedure revealed that the majority of strains are closely related. The RFLP-PGRS patterns showed a sizable group (20.5%) containing a 5.5 kb fragment and the predominant spoligotype is similar to that from the BCG vaccine strain. Unexpectedly, four strains (2.4%) showed drug resistance to 0.2 microg/ml isoniazid and 20 microg/ml ethionamide, but none of them was resistant to rifampicin or other antibiotics tested. PMID- 15876226 TI - Studies on Johne's disease in Egyptian cattle. AB - Johne's disease (JD) or paratuberculosis is a serious problem of cattle industry worldwide. For a long period of time, Egypt was considered to be free of JD. In the present study, 2150 Egyptian cattle were examined clinically for JD. Among these, samples from 160 cows were investigated for the presence of Mycobacterium avium ssp. paratuberculosis using various laboratory methods including direct microscopic examination, faecal culture and polymerase chain reaction (PCR). According to the data obtained by the culture method, positive results could be observed for 75 cows from three of five investigated districts in Egypt. Comparably investigated samples from 40 cows of one known positive flock from Hesse, Germany yielded positive reactions for 20 cows. The present study is the first description of JD in Egypt. PMID- 15876227 TI - Immunohistochemical identification of Campylobacter fetus in natural cases of bovine and ovine abortions. AB - An immunohistochemistry (IHC) procedure for the detection of Campylobacter fetus antigens using an avidin-biotin complex technique was performed on formalin fixed bovine and ovine fetal tissues from 26 natural cases of Campylobacter spp. abortion (four ovine and 22 bovine). The species of Campylobacter isolated included C. fetus ssp. venerealis from 13 bovine fetuses, C. fetus ssp. fetus from two ovine and one bovine fetus, Campylobacter jejuni from seven bovine fetuses, Campylobacter lari from two ovine fetuses and an unspeciated Campylobacter species in one bovine fetus. Histologic lesions identified in the aborted fetuses included placentitis, serositis, pneumonia, gastroenteritis, hepatitis and encephalitis. Campylobacter fetus antigens were identified by IHC in 13 of 13 bovine fetuses from which C. fetus ssp. venerealis was isolated and in two of two ovine fetuses from which C. fetus ssp. fetus was isolated. The IHC stains were negative in tissues from seven bovine fetuses from which C. jejuni was isolated, one bovine fetus infected with C. fetus ssp. fetus, one bovine fetus infected with the unspeciated Campylobacter and two ovine fetuses infected with C. lari. In positive cases, the IHC stain most frequently identified bacteria in the lung and gastrointestinal tract. The C. fetus IHC procedure performed on formalin fixed tissues is a practical tool for the diagnosis of natural cases of ovine and bovine abortion caused by C. fetus. PMID- 15876228 TI - Identification and molecular characterization of Streptococcus equi subsp. zooepidemicus isolated from camels (Camelus dromedarius) and camel milk in Kenya and Somalia. AB - Seventeen Streptococcus equi subsp. zooepidemicus strains isolated from camels and camel milk in Kenya and Somalia were identified by their cultural characteristics, by biochemical and serological reactions with the help of commercial identification systems and by molecular studies using a multiplex PCR. The isolates were further characterized by a PCR-mediated detection of size polymorphisms in the 16S-23S rDNA intergenic spacer region and the virulence gene szp and by amplification of the virulence gene cne. These molecular analysis are potentially useful in identifying and characterizing S. equi subsp. zooepidemicus strains of this origin and could possibly be valuable in epidemiological investigations. PMID- 15876229 TI - Relationship between rainfall and Neospora caninum-associated abortion in two dairy herds in a dry environment. AB - The possible direct relationship between climate variations and abortion in Neospora caninum-infected cows has not been studied. The objective of this study was to determine whether climate changes could be a risk factor for abortion in N. caninum-infected cows, and was based on yearly serological screening for neosporosis and on the confirmation of N. caninum infection on aborted fetuses in two high-producing dairy herds with a mean 27% seroprevalence of N. caninum antibodies. The final population study was comprised of 357 pregnancies in seropositive animals. Logistic regression analysis indicated no significant effects of herd, N. caninum antibody titre, climate variables during the first and third trimesters of gestation, mean and maximum temperature-humidity index values during the second trimester of gestation, and previous abortion on the abortion rate. Based on the odds ratio, a 1-unit increase in lactation number yielded a 0.85-fold decrease in the abortion rate. The likelihood of abortion was 1.9 times (1/0.54) lower for pregnant cows inseminated with beef bull semen compared with Holstein-Friesian bull semen. The likelihood of abortion decreased significant and progressively by factors of 0.5, 0.41 and 0.3 for the respective classes 40-49, 30-39 and <30 rainfall mm during the second trimester of gestation (using the class >or=60 rainfall mm as reference). As a general conclusion, it seems that increased rainfall in a dry environment can compromise the success of gestation in N. caninum-infected cows. Attempts should therefore be made to reduce environment effects during the second trimester of gestation, a period in which the immune response of cows is diminished. PMID- 15876230 TI - Books, books reviews and writing. PMID- 15876231 TI - Opening up: psychiatric nurses' experiences of participating in reflection groups focusing on the use of coercion. AB - Psychiatric staff report that the use of coercion towards patients in psychiatric care engenders feelings of discomfort and conflicts. The aim of this study was to describe psychiatric nurses' experiences of participating in reflection groups focused on the use of coercion, in relation to their views regarding systematic clinical supervision and staff support. Twenty-one nurses who had participated in reflection groups were interviewed retrospectively. The structured interview focused on: (i) their views of clinical supervision and support in general; (ii) their views of clinical supervision and support specifically concerning the use of coercion; and (iii) their experiences of participating in groups reflecting the use of coercion. Nurses were largely positive about participating in reflection groups, as expressed in the subthemes: having time for reflection; being confirmed; gaining new perspectives; sharing fellowship with colleagues; and relating more effectively to patients. Complications concerning participation in the reflection groups and other forms of clinical supervision as reported by nurses were: providing time for participation; having a common aim; being vulnerable in difficult situations; and assuming that the need for supervision and support could indicate that they were not coping with their job. PMID- 15876232 TI - Caring for clients with dual diagnosis in rural communities in Australia: the experience of mental health professionals. AB - This paper identifies and describes the experiences of 13 rural mental health professionals who care for clients diagnosed with a mental illness and a coexisting alcohol and other drug disorder (dual diagnosis). Dual diagnosis is a common problem which is often poorly understood and managed by mental health professionals. The effect of excessive substance use on a person's mental well being can present as a diagnostic challenge as each condition may mask symptoms of the other. The authors utilized a phenomenological approach to discover the experiences of a group of mental health professionals working in rural communities in Victoria, Australia. Caring for clients diagnosed with dual diagnosis was found to be a complex and stressful role that involved high levels of skill and knowledge. Despite the fact that health professionals in rural areas are expected to deliver the most appropriate care to individuals with a dual diagnosis, a number of these rural health professionals have limited preparation and experience in dealing with arising clinical diagnosis issues. Clinicians experience frustration, resentment and powerlessness in their attempt to understand their clients' drug misuse whilst simultaneously endeavouring to provide a quality mental health service. PMID- 15876233 TI - Nursing people who are suicidal on psychiatric wards in Taiwan: action/interaction strategies. AB - Suicide is a major mental health problem in Taiwan. Estimations revealed that approximately 41% of people who committed suicide had a previous history of psychiatric inpatient care. To date, a suicide nursing care theory has not been developed. Consequently, the aim of this study was to formulate a suicide nursing care theory with the aim of enhancing and advancing the nursing care provided to people who attempt suicide or have suicidal thoughts. A qualitative approach using grounded theory was adopted. A total of 15 peoples who had either suicidal ideas or had attempted suicide and 15 psychiatric nurses were interviewed and observed. Data were analysed using open, axial and selective coding and the NUD*IST software program. A substantive theory of suicide nursing care was developed from the emergent findings. Four categories surfaced in the nursing care theory relating to the nurses' 'action/interaction strategies'. They were: the holistic assessment of people who are suicidal; providing protection; providing basic care; and providing advanced care. The findings from this study could be used to influence and advance nurse education and training, clinical practice, management and further research. PMID- 15876234 TI - Whole systems working and acute inpatient psychiatry: an exploratory study. AB - The concept of 'whole system' working is fundamental to contemporary policy and consequently to practice and service development within UK mental health services. This paper reports on an exploratory study of the meaning of the 'whole system' to staff working within adult mental health services in North Wales, conducted within a broader practice development project. A focus group was used to collect data from a multiprofessional group drawn from two acute inpatient wards, Community Mental Health Teams (CMHTs) and an assertive outreach team. Thematic analysis generated three themes central to effective whole system working in adult mental health services. Discussion on the efficacy of the whole system gave rise to an understanding that this whole system varies in its effectiveness in responding to people with differing clinical presentations. Furthermore, it appears that under certain circumstances, the whole system may work to shift responsibility in order to manage worker's anxiety more adroitly than, for example, working creatively to respond to people's needs. The paper concludes that whilst by no means exhaustive or complete, these three themes enable critical reflection, discussion and identification of practice development challenges that may facilitate more effective whole system working and better services in North Wales and possibly in other settings. PMID- 15876235 TI - Adverse factors and the mental health of older people: implications for social policy and professional practice. AB - Defining 'older people' as a homogenous group is problematic; it can lead to stereotypical and stigmatizing perceptions of what old age is, attracting consequent negative attitudes to later life. Nevertheless, evidence suggests that some in the older age bracket are subject to particular stressors and physical changes that can adversely affect their mental health. This paper will consider challenges to mental health in older age groups and particularly the phenomenon of dementia. The role and influence of diagnosis, social policy and professional practice will also be addressed and suggestions will be made as to how people could improve their responses to either the predisposition to or the actual occurrence of mental distress in later life. In addition, it is argued that person-centredness is important as the caring/cultural medium through which provisions and policies are mediated: that obtaining appropriate balances between corporate and individual contributions and interventions must constitute the context wherein future developments lie. PMID- 15876236 TI - Nurses' perceptions of the working environment: a UK independent sector study. AB - The working environment is an important determinant of employee well-being. Previous UK studies report registered nurses' perception of the working environment using the Work Environment Scale (WES), but surprisingly few include data for nurses working in mental health or learning disability settings. One hundred and sixty-one (58.8%) registered nurses working day shifts in five specialist divisions (forensic adult mental health, adolescent mental health, elderly continuing care, adult and adolescent learning disability and brain injury rehabilitation) of a large charitable hospital comprising mostly secure (i.e. 'locked') wards completed the WES. Nurses working in separate clinical divisions differed only on the 'physical comfort' subscale. These results are discussed in the context of previous UK research: this sample of nurses scored differently on a number of subscales, with the working environment characterized by relatively high levels of support, cohesion and managerial control and slightly lower levels of autonomy. It is unclear whether the differences reflect the organizational (i.e. non-NHS) context, or a secure environment effect. Previous studies of mental health nurses are now 10 years old and we present data that may provide a benchmark of perceptions of the working environment held by nurses working in mental health and learning disability settings. PMID- 15876237 TI - Coping with uncertainty within the preceptorship experience: the perceptions of nursing students. AB - A substantial amount of time and resources are channelled into supporting clinical practicum in nursing education programmes. Attention is targeted at the most effective models to achieve this aim. The provision of sound support models regardless of specific clinical setting is recognized as integral to student development and transition periods throughout nurses' careers. Within the mental health setting, this situation is compounded by the negative attitudes nursing students tend to hold towards people experiencing a mental illness. Preceptorship has been widely used both nationally and internationally for clinical practicum. Although this model seems to have been endorsed by virtue of its increasing use, additional examination is necessary to assess efficacy and effectiveness across clinical practicum, including those in mental health settings. In utilizing a grounded theory approach, this study addresses the question: what is the experience of preceptorship for undergraduate nursing students in the mental health setting? The major themes identified include: 'fear of the unknown', 'reconciling difference between general acute health and mental health settings' and 'supporting practice'. PMID- 15876238 TI - The clinical activity of mental health nurse lecturers. AB - Concerns have been expressed about the ability of mental health lecturers to prepare practitioners with the skills, knowledge and attitudes needed to implement the National Service Framework for mental health. Specifically mental health nurse lecturers have been criticized for lacking clinical competence and credibility, and for being out of touch with developments in service delivery. This paper presents a selection of findings from a recent review into the clinical activity of mental health lecturers in higher education institutions in England. The issues discussed include roles in practice, questioning the value of face-to-face clinical work, the benefits of being clinically active, the difficulties of clinical academic roles, barriers to maintaining clinical activity, questioning the quality of the clinical workforce, and finally strategies to facilitate clinical activity. Whilst there is support among mental health nurse lecturers for the need to engage with practice at some level, there are inconsistencies in the ways that this is being achieved. Lecturers are, however, getting together to debate how best to keep up with changes in mental health practice and delivery. PMID- 15876239 TI - The powers and duties of psychiatric nurses under the Mental Health Act 1983: a review of the statutory provisions in England and Wales. AB - This paper examines the powers and duties that psychiatric nurses have under the Mental Health Act 1983 with respect to the care and treatment of mentally disordered people. This statute saw for the first time specific reference to the powers and duties of psychiatric nurses. These powers and duties are primarily concerned with the nurse's role in relation to consent to treatment, the administration of medication, the right to be consulted and to ensuring that detained patients are informed of their rights under the 1983 Act. The 1983 Act also provides and extends the powers of psychiatric nurses to detain certain inpatients with mental disorder against their will. In order to avoid the potential charges of 'treatment without consent' and 'unlawful detention', it is vital that psychiatric nurses, when caring for this client group, have a sound working knowledge of their powers and duties under current legislation. The legal definitions of mental disorder are explored as well as the issue of 'treatability' for mental disorder in clinical practice. PMID- 15876240 TI - The Impact Message Inventory (IMI-C): generalizability of patients' command and relationship messages across psychiatric nurses. AB - This paper tests the hypothesis that patients' stereotypical dysfunctional interpersonal communication styles would be validly measurable by the command or relationship messages experienced by psychiatric nurses while interacting with the patient in the relatively unstructured every day context of the psychiatric ward. The generalizability of interpersonal pressures exerted by patients was tested by having different psychiatric nurses independently appraise patients' command or relationship messages with the Impact Message Inventory, circumplex version (IMI-C). Generalizability of interpersonal pressures strongly depended on the specific combination of psychiatric nurses. Out of eight classes of command or relationships three turned out to be generalizable across nurses: Dominance, Hostile-Submissive and Friendly Dominance. PMID- 15876241 TI - Mothers talk about their children with schizophrenia: a performance autoethnography. AB - This article presents the script for a performance autoethnography that explores how mothers of adults with schizophrenia talk about their children. The script draws on my own experience as a mother of a person with schizophrenia, interviews I conducted with mothers of people with schizophrenia, and written accounts by mothers of people with schizophrenia. Autoethnography refers to an approach to social science research in which the experience of the researcher is recognized as a salient part of the research process. Autoethnography typically uses non traditional genres, such as this script, for presenting research to address ethical and political problems in the representation of research. These genres allow researchers to both break with and continue the ethnographic tradition of representing the lives and experiences of others. PMID- 15876242 TI - Improving patient access and choice: Assisted Bibliotherapy for mild to moderate stress/anxiety in primary care. AB - Current traditional methods of mental healthcare service delivery, based on 'specialists' providing 'outpatient appointments' for formal therapy, are often inappropriate for the needs of patients in primary care. The estimated numbers of adults with mental health problems are immense, and it is this, combined with Department of Health initiatives aimed at improving choice and access, which make it essential that new ways of delivering services are explored. This trial examines the use of an assisted self-help treatment package for mild to moderate stress/anxiety [Assisted Bibliotherapy (AB)] with an adult clinical population referred by their general practitioner. Assisted Bibliotherapy is a brief intervention (8 weeks), with limited therapist contact (20-min sessions). Non parametric statistical testing of scores from the Zung Anxiety Scale and the Clinical Outcomes in Routine Evaluation (CORE) questionnaire indicated positive results. There was significant improvement at post-treatment, which was maintained at 3 month follow-up. The results from this trial and a previous trial of AB by Kupshik & Fisher in 1999, indicate that it is an effective treatment which could be used as part of a stepped care approach to managing and treating stress/anxiety in primary care. PMID- 15876243 TI - Work stress and physical assault of nursing aides in rural nursing homes with and without dementia special care units. AB - PURPOSE: This study compared nursing aides (NAs) employed in rural nursing homes with and without dementia special care units (SCUs) on (1) exposure to and distress from disruptive behaviours exhibited by residents, (2) job strain and (3) physical assault. DESIGN AND METHODS: The data were drawn from a larger study conducted in Saskatchewan, Canada, in which all rural nursing homes of < or = 100 beds that had an SCU were matched to same-sized rural facilities with no SCU. Nursing aides (n = 355) completed a mailed survey questionnaire. RESULTS: Nursing aides employed in nursing homes with an SCU reported significantly less frequent exposure to disruptive behaviours (including aggressive and aversive behaviours) than NAs in non-SCU facilities, less distress when these behaviours were directed toward them, less exposure to aggressive behaviour during caregiving, lower job demands and lower job strain. There was a trend toward increased risk of being assaulted in the last year associated with being in a non-SCU facility. Having a permanent position, increased job strain, and feeling inadequately prepared for dementia care were significantly associated with higher risk of being assaulted. In the SCU facilities, NAs who worked more time on the SCU reported more assaults but less distress from disruptive behaviour, lower psychological job demands, lower job strain and greater work autonomy. IMPLICATIONS: Providing more dementia care training and reducing job demands and job strain may help to reduce work related stress and physical assault of nursing aides employed in nursing homes. PMID- 15876244 TI - An exploration of the decision-making processes at the point of referral to an Admiral Nurse team. AB - The Admiral Nursing Service supports individuals caring for people with dementia. As a response to the high demand for one team's service, a temporary closure to referrals was agreed. This provided the impetus to explore the influence of nurses' decision making at the point of referral on demand for the service. The research questions addressed were: How do Admiral Nurses make decisions regarding referrals and What factors influence this decision-making process? Sixteen case studies were retrospectively reviewed through the auspices of a semi-structured interview aimed at identifying which factors influenced the nurses' acceptance or rejection of these referrals. Results indicated that the Admiral Nurses had a clearly identified focus in supporting the carers of people with dementia, but that the complexity of the carers situation which included the person with dementia was significant. However, the desire to fulfil a case-management role while attempting to provide a service that is of a specialist nature and of limited capacity generated tensions. The decision to offer a service to carers was consequently influenced not only by perceived need but also upon the nurses feeling professionally responsible for perceived gaps in service provision. PMID- 15876245 TI - Childhood abuse in Swedish female users of psychiatric services. AB - INTRODUCTION: The aims of this study were to investigate: (1) the prevalence of childhood abuse in women admitted to psychiatric services in a county in the south of Sweden; (2) who the perpetrators were; and (3) the women's self-reported consequences of childhood abuse. METHOD: The study had a cross-sectional design and was a part of a more comprehensive study. An anonymous self-reported questionnaire was used which included both closed and open-ended questions. The data material were analysed by means of descriptive statistics, Mann-Whitney U test and manifest content analysis. RESULTS: The total number of women who participated in the study was 259, 51% of whom reported experiences of abuse during childhood, with 53% of these having been exposed to more than one type of abuse. The most frequent perpetrator was the woman's parents; mainly the fathers but also the mothers turned out to be frequent perpetrators of abuse. Some 75% of the women reported current psychological problems in adulthood related to abuse in childhood. According to a manifest content analysis, five themes of self reported psychological problems emerged: psychiatric problems, shortcomings in social relations, poor self-confidence, fears and bad memories. PMID- 15876246 TI - Action research: the revision of services at one mental health rehabilitation unit in the north of England. AB - This paper reports the experiences of people in one mental health rehabilitation unit in the north of England, where conventional individualized approaches have repeatedly failed, leading to bed blocking and inertia. An action research approach has been used to identify the key problems concerned, construct appropriate goals and formulate problem-solving plans, leading to the development of an alternative therapeutic regime. The new facility is based on a society centred, recovery approach which emphasizes the principles of community involvement, social responsibility and meaningful occupation of time. This approach may be of transferable interest to others in the field. The paper outlines the history of the unit, describes the action research process, and presents the philosophy of care, nursing model, admission criteria and main assessment tool now being used. It concludes with an open review of the research process, exploring both positive and negative aspects. PMID- 15876247 TI - Review of seclusion policies in high secure hospitals and medium secure units in England, Scotland and Wales. PMID- 15876249 TI - Microarray-based classification of diffuse large B-cell lymphoma. AB - OBJECTIVE: Hierarchical clusterings of diffuse large B-cell lymphoma (DLBCL) based on gene expression signatures have previously been used to classify DLBCL into Germinal Center B-cell (GCB) and Activated B-cell (ABC) types. To examine if it was feasible to perform a cross-platform validation on the Affymetrix HG-U133A oligonucleotide arrays and improve the classification, we determined the expression profiles of pretreatment, diagnostic samples from 52 primary nodal DLBCL. METHODS AND RESULTS: First, three previously published gene lists were converted to the HG-U133A probe sets and used for hierarchical clustering. In this way, three subtypes, including the GCB type (n = 20), the ABC type (n = 25) and an intermediate group, Type-3 (n = 5), were distinguished. The CD10 and Bcl-6 expression as well as t(14;18) translocation were prevalent, but not exclusive to the GCB type. By contrast, MUM1 was only expressed in the ABC and in Type-3 samples. The 5-year survival was similar between the groups, but GCB patients showed a better initial response to CHOP or CHOP-like regimens than the remaining patients and tended to have less advanced disease and lower IPI scores. As a next step, an improved set of classifier genes was generated by analysis of 34 patients that were consistently classified as GCB or ABC in the above analyses. Seventy-eight genes were selected and demonstrated on two previously published data sets (Shipp et al. Nat Med 2002;8:68-74 and Houldsworth et al. Blood 2004;103:1862-1868) to exhibit a higher specificity than the original gene lists. CONCLUSION: We conclude that gene expression profiling with Affymetrix Genechips is efficient to distinguish between GCB and ABC types of DLBCL and that these are likely to represent separate biological entities. The Genechip platform is highly standardised and therefore useful for future prospective investigations to establish the value of gene expression profiling in the clinical management of DLBCL. PMID- 15876250 TI - Children and adults with acute lymphoblastic leukaemia have similar gene expression profiles. AB - OBJECTIVES: To compare the gene expression pattern in children and adults with acute lymphoblastic leukaemia (ALL) in order to improve our understanding of the difference in disease biology and prognosis. METHODS: The gene expression profiles in diagnostic samples from 29 children and 15 adults with ALL were analysed using the oligonucleotide chip Hu95ver2a, produced by Affymetrix. RESULTS: Unsupervised hierarchical cluster analysis revealed that, in spite of differences in outcome, patients clustered irrespective of age, first by T-cell or B-precursor immunophenotype, and second by cytogenetic changes within the B precursor group. The expression pattern analysis allowed the reclassification of some samples into the proper cytogenetic group. We also showed that separate clustering of samples with the BCR/ABL translocation could be explained by different breakpoint regions in the BCR. No significant difference in gene expression was observed between samples with and without CDKN2A deletion within the B-precursor group. Analysis of different age groups revealed a similarity in expression profiles when infants with the MLL translocation and adults over 40 yr of age were compared irrespective of karyotype. CONCLUSIONS: In spite of the difference in clinical outcome, the gene expression pattern in children and adults with ALL is very similar and is primarily dependent on immunophenotype and cytogenetic aberrations. However, when age groups are compared, the expression patterns of infants and adults over 40 show a remarkable similarity. PMID- 15876251 TI - Expression of DNA repair gene Ku80 in lymphoid neoplasm. AB - OBJECTIVES: Ku, a heterodimer of KU70 and Ku80 that binds to double-strand DNA breaks (DSBs) and activates the catalytic subunit (DNA-PKcs) when DNA is bound, is essential in DSB repair and V(D)J recombination. Ku80 is a putative tumor suppressor gene that might play an important role in drug resistance. Our aim was to determine the role of Ku80 in lymphoid malignancy. PATIENTS AND METHODS: Competitive reverse transcription-polymerase chain reaction assays were performed and the expression levels of Ku80 were measured in normal peripheral blood mononuclear cells (n = 9) and malignant cells from 25 patients with acute lymphoblastic leukemia (ALL) (14 children, 11 adults), and chronic lymphoproliferative disorders (n = 6). The Ku80 transcripts were sequencing for the possibility of mutation. RESULTS: No mutation or Ku80 variant at the RNA level was seen in any patient samples or in the Raji or CCRF-CEM cell lines. In Ku80 expression, 8.8-, 1.9-, and 6.2-fold mean increases were seen in adult, pediatric ALL, and chronic lymphoid malignancies compared with the control. The Ku80 was significantly higher in adult than in pediatric ALL (P = 0.02). The amount of Ku80 expression in ALL was moderately correlated with peripheral white blood cell counts, but not with Ki67 labeling index. High Ku80 expressers (higher than the mean of all patients with ALL) tended to respond poorly to therapy: Only 22% of high Ku80 expressers achieved durable complete remission compared to 62% of low expressers. CONCLUSIONS: Our study suggests that Ku80 might contribute to generally poor prognoses in adult ALL. PMID- 15876252 TI - Management of patients with polycythaemia vera: results of a survey among Swedish haematologists. AB - The prevailing attitudes regarding diagnostic and therapeutic procedures in patients with polycythaemia vera (PV) among Swedish haematologists were surveyed by way of a mailed questionnaire in August 2002. Among diagnostic procedures frequent use is reported for arterial O(2) saturation, spleen size determination, bone marrow histology, serum erythropoietin, serum cobalamins and leukocyte alkaline phosphatase score, while direct determination of the red blood cell mass is used infrequently (seldom or never by 82%). Among therapeutic modalities hydroxyurea and phlebotomy alone were most frequently used. The (32)P therapy was used at least sometimes by 57% of the physicians, and more widely in the university clinics. Anagrelide and alfa-interferon was used in a minority of patients only. The use of prophylactic acetylsalicylic acid was very variable. The majority of the physicians had an aim for their phlebotomy treatment at a level of 0.45 or less, but 21% used a level of 0.46-0.49 and 8% a level of 0.55 0.60 (in younger patients). The platelet level, at which myelosuppressive therapy was initiated, also varied, from 400 x 10(9)/L to >1500 x 10(9)/L. It can be concluded that in practical clinical work in Sweden the diagnosis of PV is established by frequent use of serum erythropoietin, bone marrow examination and spleen size determination. The use of different therapeutic modalities is very variable. Many physicians carry out their phlebotomy treatment with less intensity compared with national and international recommendations. PMID- 15876253 TI - The bisphosphonate zoledronic acid inhibits the development of plasmacytoma induced in BALB/c mice by intraperitoneal injection of pristane. AB - OBJECTIVES: Bisphosphonates (BPs) are mostly used in the palliative care of myeloma-associated osteolytic lesions. Recent studies have suggested that BPs may also exert direct antitumor effects on myeloma cells. We have investigated the effect of the potent bisphosphonate, zoledronic acid (ZOL), on the development of pristane (2,6,10,14-tetramethylpentadecane)-induced plasmacytoma (PCT) in six week-old BALB/c mice. METHODS: Different groups of pristane-treated mice also received ZOL (100 microg/kg) commencing after the development of PCT or ZOL (20 microg/kg) from the first day. Control groups received pristane alone, ZOL alone (20 microg/kg), or phosphate-buffered saline. The study was terminated on day 300, and the remaining mice were autopsied and abdominal tissues were examined histologically for PCT. RESULTS AND CONCLUSIONS: Statistical analysis revealed a significant delay in PCT development in the group receiving pristane plus ZOL (20 microg/kg) from the first day compared to the groups receiving pristane alone and pristane combined with ZOL (100 microg/kg) after the appearance of PCT (Log-rank, P = 0.0001 and 0.0001; respectively). Kaplan-Meier analysis revealed a significant difference in survival between the group treated with pristane alone and the groups receiving pristane plus ZOL (20 microg/kg) from the first day or ZOL (100 microg/kg) after the appearance of PCT (Log-rank, P = 0.016 and 0.023; respectively). These results indicate a direct anti-tumor effect of ZOL in pristane-induced PCT development BALB/c mice, which may contribute to their significantly increased survival. This hypothesis should now be further investigated in clinical trials. PMID- 15876254 TI - Grading of symptoms in hyperleukocytic leukaemia: a clinical model for the role of different blast types and promyelocytes in the development of leukostasis syndrome. AB - OBJECTIVE: Patients with hyperleukocytic leukaemia were graded according to the severity of symptoms possibly caused by leukostasis to evaluate the effectiveness of therapy and to test the relative contribution of blast type and count of blasts and promyelocytes in the development of leukostasis syndrome. METHODS: Ninety-five patients (59 male, 36 female, median age 52 yr) with hyperleukocytic leukaemia [leukocytes above 50 x 10(9)/L, 48 acute myeloid leukaemia (AML), 31 chronic myeloid leukaemia (CML), 13 acute lymphoblastic leukaemia (ALL), three chronic myelomonocytic leukaemia (CMML)] were grouped according to the presence or absence and severity of neurologic, pulmonary and other symptoms into four categories (no, possible, probable and highly probable leukostasis syndrome). Age, white blood count (WBC), haemoglobin, blast count and total of blasts plus promyelocytes of these groups were compared by Mann-Whitney U-test. RESULTS: Patients with myeloid leukaemia (AML M1/M2, CML) which scored as highly probable leukostasis showed significantly higher WBC (P = 0.011), lower haemoglobin (P = 0.004), higher peripheral blast counts (P = 0.004) and higher total of peripheral blasts plus promyelocytes (P < 0.001) compared with the lower probability groups. In leukaemia involving the monocytic lineage (AML M4/M5, CMML) no significant differences were found in any of these factors between patients with highly probable leukostasis and the other patients. CONCLUSIONS: Our results show that a four-stage clinical grading scale is a valuable tool for analysing hyperleukocytic patient populations and evaluate the effectiveness of therapy more precisely. We further demonstrate that the mechanisms of leukostasis are different in myeloid leukaemia as compared with leukaemia with involvement of the monocytic lineage. PMID- 15876255 TI - Sequential organ failure predicts mortality of patients with a haematological malignancy needing intensive care. AB - OBJECTIVES: Poor survival of patients with a haematological malignancy admitted to the intensive care unit (ICU) prompts for proper admission triage and prediction of ICU treatment failure and long-term mortality. We therefore tried to find predictors of the latter outcomes. METHODS: A retrospective analysis of charts and a prospective follow-up study were done, of haemato-oncological patients, admitted to our ICU in a 7-year period with a follow-up until 2 yr thereafter. Clinical parameters during the first four consecutive days were taken to calculate the simplified acute physiology (SAPS II) and the sequential organ failure assessment (SOFA) scores, of proven predictive value in general ICU populations. RESULTS: From a total of 58 patients (n = 47 with acute myelogenous leukaemia or non-Hodgkin lymphoma), admitted into ICU mostly because of respiratory insufficiency, sepsis, shock or combinations, 36 patients had died during their stay in the ICU. Of ICU survivors (n = 22), 20 patients died during follow-up so that the 1-year survival rate was only 12%. The SAPS II and particularly the SOFA scores were of high predictive value for ICU and long-term mortality. CONCLUSIONS: Patients with life-threatening complications of haematological malignancy admitted to ICU ran a high risk for death in the ICU and on the long-term, and the risk can be well predicted by SOFA. The latter may help us to decide on intensive care in individual cases, in order to avoid potentially futile care for patients with a SOFA score of 15 or higher. PMID- 15876256 TI - Early detection of cardiac dysfunction in thalassemic patients by radionuclide angiography and heart rate variability analysis. AB - BACKGROUND: Cardiac dysfunction remains the major cause of death in beta thalassemia. Aim of this study was to assess early myocardial damage in thalassemic patients with no symptoms or echocardiographic evidence of dysfunction at routine monitoring. METHODS: Twenty patients (seven females; median 25 yr [first quartile 22,third quartile 28]) with beta-thalassemia underwent radionuclide angiography (RNA) at rest and during low-dose dobutamine infusion (5-10 gamma/kg/min). Right and left ventricular ejection fractions (EF) were determined by first-pass method and gated equilibrium acquisition, respectively. Twenty-four-hour Holter monitoring with time-domain heart rate variability (HRV) assessment and echocardiographic follow-up (21 months [5,27]) were performed. RESULTS: Eleven patients showed regional wall motion abnormalities at RNA; left ventricular EF, HR and diastolic measurements significantly increased after dobutamine infusion. Patients with abnormal RNA right ventricular EF (n = 8, <0.45) showed lower echocardiographic left ventricular EF at the enrollment (0.54 [0.50,0.61] vs. 0.62 [0.56,0.67], P = 0.02) than those with a normal right ventricular EF. Patients with reduced standard deviation of the averages of RR intervals in all 5-minute periods of entire recording (SDANN) (n = 6, <100 ms), a measure of HRV, had lower echocardiographic left ventricular EF (0.53 [0.49,0.62] vs. 0.62 [0.56,0.66], P = 0.03) and lower fractional shortening (0.28 [0.25,0.32] vs. 0.36 [0.30,0.39], P = 0.003) at the enrollment than those with normal SDANN. No significant association was found between RNA and HRV measurements and follow-up left ventricular function. CONCLUSIONS: Right ventricular dysfunction and abnormal HRV may represent the early features of cardiac disease in thalassemic patients with no evidence of ventricular dysfunction at routine evaluation. PMID- 15876257 TI - Long-term treatment with deferiprone in a L1 veteran. AB - We studied a patient with mild beta-thalassaemia major under treatment with the oral chelator deferiprone (DFP or L1) for about 10 yr (L1 veteran). Due to poor compliance with desferrioxamine, the patient started compassionate use of DFP at an age of 23 yr with a serum ferritin of 5200 microg/L. Monitoring iron overload by SQUID biosusceptometry revealed a dramatic decrease of liver iron concentrations from 4500 to 950 microg/g(liver) within 9.5 yr. A good clinical response to chelation treatment with DFP was observed together with an improvement of liver and cardiac function and a reduction in the hepatitis virus load. PMID- 15876258 TI - Effective in vivo purging with rituximab and autologous peripheral blood stem cell transplantation in a woman with CD5 positive primary cutaneous diffuse large B-cell lymphoma. AB - Generalized subcutaneous tumors developed without any other sites of the disease in a Japanese woman. Skin biopsy revealed CD5(+) and CD20(+) atypical diffuse large cells infiltrating subcutaneous tissues. The diagnosis was CD5(+) primary cutaneous diffuse large B-cell lymphoma. Tumor-specific PCR showed the existence of malignant cells in the peripheral blood and bone marrow. After three cycles of chemotherapy, she was remained in partial remission. Peripheral blood stem cells (PBSC) were harvested after the fourth cycles of chemotherapy combined with rituximab for in vivo purging. The contamination of tumor cells in PBSC was negative with PCR. She then underwent autologous peripheral blood stem cell transplantation using purged PBSC and has remained in complete remission for the past 24 month. PMID- 15876259 TI - Successful non-T-cell-depleted HLA-haploidentical 3-loci mismatched bone marrow transplantation. AB - A 17-year-old boy with therapy-related acute myelocytic leukemia (FAB classification-M0) successfully received allogeneic non-T-cell depleted (non-TCD) bone marrow transplantation (BMT) from his 3-loci HLA-mismatch mother, although pre-BMT detection of feto-maternal microchimerism was negative. The BMT was performed with reduced intensity conditioning (total body irradiation; 4 Gy, fludarabine; 20 mg/m(2) x 6, and melphalan; 70 mg/m(2) x 2) and short-course methotrexate and tacrolimus for GVHD prophylaxis. Complete donor chimera was obtained on day 19, associated with Grade 3 acute GVHD (skin: Stage 1, liver: Stage 0, gut: Stage 3) that was well controlled with immunosuppressive therapies. At day 200 of transplantation, he was in complete remission with no signs of chronic GVHD. Our case suggests that non-TCD HLA-haploidentical 3-loci mismatched BMT can be safely performed from mother to offspring even when feto-maternal microchimerism is barely detectable with the current detection procedure. PMID- 15876260 TI - Dialysate concentration and pharmacokinetics of 2F-Ara-A in a patient with acute renal failure. AB - Fludarabine is frequently used for treatment of B-cell chronic lymphocytic leukemia and in conditioning regimes for hematopoietic cell transplantations. The total body clearance of the principal metabolite 2-fluoro-ara-A (2F-Ara-A) correlates with the creatinine clearance. We report data on total dialysate concentration as well as pharmacokinetics of 2F-Ara-A in a patient with anuric acute renal failure. On three consecutive days the patient received a daily dose of 80 mg (40 mg/m(2)) fludarabine and underwent three consecutive extended (daily) dialysis (ED) sessions. ED removed a considerable amount of the drug. The average dialysis clearance was 33.85 ml/min which is about 25% of the clearance in patients without renal failure. No toxic side effects of the treatment were observed. This case suggests that fludarabine treatment can be considered in patients requiring dialysis if dose reduction and adequate removal of the drug by hemodialysis is provided. PMID- 15876261 TI - Imatinib in pregnancy. PMID- 15876262 TI - Correlation of idiopathic thrombocytopenic purpura (ITP) with anti-microsomal antibody. PMID- 15876263 TI - Main clinical manifestations of a bleeding diathesis: an often disregarded aspect of medical and surgical history taking. AB - A suitable clinical evaluation of a bleeding diathesis is often forgone. The young doctor is often unprepared to describe in an accurate way the different types of bleeding. An adequate classification and adequate clinical information about a bleeding diathesis are instead of paramount importance. Bleeding may be cutaneous, mucous, articular, muscular, parenchymal, intracavitary, orificial. Each of these sites and forms may have diagnostic implications. An accurate description of the several forms of cutaneous bleeding (petechiae, purpuric spots, ecchymosis, haematomas, etc.) is needed for referrals and for controls. The correct evaluation of cutaneous bleeding manifestations of children (battered child syndrome) is absolutely important for clinical and medico-legal purposes. The same is true for the battering syndrome seen in women abused by their spouses. The grading of haemarthrosis in haemophilia patients is important for the follow-up. A proper description of haematuria is essential in suggesting the probable site of bleeding (kidney or bladder or urethra). A proper evaluation of bleeding may give also useful information on the general health status of the patients (presence of anaemia, poor nutrition, renal insufficiency, etc.). The combination of bleeding and thrombosis in the same patient is also a clinical challenge. The relationship between haemorrhage and thrombosis may be sequential or concomitant. Sequential thrombosis may occur in a patient confined in bed for a brain haemorrhage. Concomitant thrombosis and bleeding occur in DIC and in patients with thrombosis being treated with anticoagulants. Finally, it should be kept in mind that a proper evaluation of the bleeding diathesis of a given patient may help the caring doctor in ordering appropriate laboratory tests (e.g. a platelet count for petechiae, a PTT for a patient with haemarthrosis, etc.). PMID- 15876264 TI - Prophylactic recombinant factor VIIa in haemophilia patients with inhibitors. AB - Prevention of bleeding, especially into joints, with prophylactic factor infusions is the most effective treatment for severe haemophilia patients. Approximately 15-30% of patients with factor VIII deficiency and 3-5% of patients with factor IX deficiency develop neutralizing antibodies (inhibitors) to factor precluding their use. Such patients often have significant bleeding complications including life- and limb-threatening bleeds and severe joint disease. Prophylaxis for such patients is not generally considered because of the fact that the standard (bypassing) agents for such patients are not as effective as natural factor replacement, because of concerns for thrombotic complications and also because of the very high cost of bypassing agents. We treated two patients with high titre inhibitors with prophylactic recombinant factor VIIa (rFVIIa). The first patient was treated as a result of development of a target joint and to reduce the use of agents that can lead to anamnesis of his inhibitor. The second patient had multiple severe bleeds and was hospitalized 20% of the time over a 2 year period. He had a very poor quality of life. Both patients had shown good responses previously to rFVIIa for treatment of bleeds. Both patients had an outstanding response to prophylaxis albeit at a very high cost. Prophylaxis with rFVIIa can be an effective approach in select inhibitor patients with severe complications related to bleeding. PMID- 15876265 TI - Retrospective audit of a continuous infusion protocol for haemophilia A at a single haemophilia treatment centre. AB - Continuous infusion (CI) of factor concentrates has been suggested to decrease the risk of bleeding and reduce cost in the treatment of bleeding disorders. Concerns have also been raised regarding stability and sterility of products administered by CI, the risk of local thrombophlebitis and an association with the development of an inhibitor in mild haemophilia. A retrospective chart review was conducted to investigate a CI protocol regarding product use, maintenance of FVIII levels and the frequency of complications including inhibitor development. Twelve patients with haemophilia A received recombinant factor VIII by CI a total of 18 times between April 1998 and September 2003. Ages ranged from 4 months to 75 years and indications for treatment included severe bleeds and surgical prophylaxis. The protocol which was audited required a bolus of 50 U kg(-1) of FVIII followed by CI at an initial rate of 4 U kg(-1) hr(-1). All infusions were administered by i.v. infusion after diluting the reconstituted concentrate in saline. There were no documented cases of significant bleeding, adverse reactions, thrombophlebitis or infection. Two mild haemophilia A patients developed a low titre inhibitor after receiving CI. It is not clear in either case that CI was the main contributing factor. Our CI protocol will now be modified to use less product, aiming for more cost-effectiveness. PMID- 15876266 TI - Handheld computers and paper diaries for documenting the use of factor concentrates used in haemophilia home therapy: a qualitative study. AB - A recently published randomized controlled trial (RCT) showed that adherence to infusion diary record keeping was improved by the use of handheld computers. In this study, attitudes to record keeping were explored and patient preferences regarding the method of recording determined for the patients who participated in the trial. Qualitative study consisting of individual semi-structured interviews with 20 severely affected patients with haemophilia who participated in an RCT. Individuals were purposefully sampled based on their recent method of record keeping and whether child or adult. Analysis employed a constant comparative method to identify key themes from the data. Most individuals (19 of 20, 95%) considered record keeping to be important. They readily identified reasons to keep records: to benefit themselves, their families, clinical staff, product distributors and manufacturers. Keeping records helps them: feel a part of the health care team; have confidence they would be notified of product recalls; review their past history; improve their ability to advocate for themselves and improve communication among all parties. Record keeping, particularly when using paper diaries, can be burdensome and a challenge to maintain consistently. All 10 individuals (100%) who had used both paper diaries and handheld computers preferred the latter. Most patients understand that record keeping can be of benefit to them. Clinics can use this knowledge to inspire other patients by developing educational programmes that de-emphasize authority. In addition, given the evidence of both patients' preference for handheld computers, and the effectiveness of this approach documented in an RCT, switching to handheld computers is likely to improve record keeping. PMID- 15876267 TI - Synovitis in a murine model of human factor VIII deficiency. AB - Recurrent joint bleeding is the most common musculoskeletal manifestation of haemophilia and leads to a target joint and synovitis. The pathobiology of haemophilic synovitis (HS) is not well understood. Here the histopathological changes that occur following haemarthrosis were examined in an animal model for human HS. After two haemarthrosis, there was soft tissue and joint swelling and histological changes of acute synovitis included infiltration of the sub-synovial layer by mononuclear cells and neutrophils, thickening of the synovial membrane with villus formation, and hyperplasia of blood vessels. Subacute changes were evident after three haemarthrosis; muscle atrophy was present and an intense mononuclear cell infiltrate filled the sub-synovial space. There was destruction of articular surfaces and loss of cartilage. Seventeen months after three haemarthrosis, chronic joint changes included gross deformity and loss of congruence due to dense fibrotic tissue filling the joint space. The mononuclear inflammatory cell infiltrate and thickened synovial membrane persisted. Pits and erosions of articular surfaces and sub-chondral cysts were present. There was fibro-cartilage and new bone formation. This model of human HS should be useful to fully evaluate the biochemical and molecular changes that occur following joint bleeding and to test novel therapeutics to prevent HS. PMID- 15876268 TI - Knee and hip arthroplasty infection rates in persons with haemophilia: a 27 year single center experience during the HIV epidemic. AB - Total joint replacement (TJR) is an option for the management of chronic haemophilic arthropathy. Because surgery is technically challenging, there is a high rate of deep prosthetic infections, particularly in human immunodeficiency virus (HIV)-infected individuals. We determined the incidence of deep infection rates following total knee and hip arthroplasties in HIV-seropositive and HIV seronegative persons with haemophilia. Fifty-one primary joint replacements were performed on 32 patients seen at a regional comprehensive haemophilia care center from 1975 to 2002. Thirty prostheses were placed in patients who were HIV seropositive prior to surgery (n = 14) or seroconverted later (n = 16). Median age at the time of surgery was 33 years (range: 20-61) among 19 HIV-seropositive patients and 35 years (range: 26-74) among 13 HIV-negative patients. Median duration of follow-up was 83 months (range: 2-323). Rate of primary joint infection per artificial joint-year by HIV status was compared by Poisson regression. Main outcome measures were the incidence of primary replacement joint infections by HIV status. Deep infections developed in five (9.8%) of 51 replacement joints. There were two infections during 204.15 joint-years without HIV infection and three infections during 205.28 joint-years with HIV infection. The incidence rate of joint infection (0.98 vs. 1.46 per 100 joint-years) was not increased with HIV (relative risk, RR: 1.49, 95% CI: 0.25-8.93, P = 0.66). We conclude that HIV infection is not a contraindication to knee or hip replacement arthroplasty in the appropriate clinical setting. PMID- 15876269 TI - Rofecoxib as adjunctive therapy for haemophilic arthropathy. AB - Joint haemorrhage and subsequent haemophilic arthropathy are significant complications in haemophilia. The pathophysiology involves inflammation and angiogenesis. Cyclooxygenase-2 (COX-2) inhibitors are anti-inflammatory agents, which have potent anti-inflammatory, anti-angiogenic and analgesic properties yet do not affect platelet function in the manner of traditional non-steroidal anti inflammatory drugs. These properties make such agents potentially useful as adjunctive therapy in haemophilia. There is only one prior report describing rofecoxib treatment in a single haemophilia patient. Our objectives were to determine the safety and efficacy of rofecoxib in treating acute haemarthrosis, chronic synovitis, target joints and pain. We conducted a retrospective medical record review of patients treated with rofecoxib for acute haemarthrosis, chronic synovitis, target joint or pain. The safety and efficacy of rofecoxib treatment were determined based on subjective patient reports and physical examinations during follow-up clinic visits. A total of 28 patients between 3 and 37 years of age were treated for a total of 42 courses of rofecoxib treatment. All courses were evaluated for safety and 31 for efficacy. Rofecoxib was used for eight acute haemarthrosis, four target joints, seven cases of synovitis and 12 episodes of pain. Efficacy was demonstrated particularly for chronic synovitis and pain and no serious adverse events occurred. This is the largest study to date evaluating COX-2 inhibitors as adjunctive therapy in haemophilia and suggests that these agents may be an important adjunctive therapy in the management of haemophilia. PMID- 15876270 TI - Reliability of progressive and additive MRI scoring systems for evaluation of haemophilic arthropathy in children: expert MRI Working Group of the International Prophylaxis Study Group. AB - Effective treatment of haemophilic arthropathy requires a detailed evaluation of joint integrity. Methodological assessment of magnetic resonance imaging (MRI) scores are needed to assure reproducibility of measurements when comparing results of clinical trials conducted in different centres. We compared the reliability of two MRI scoring systems for assessment of haemophilic arthropathy: one progressive system that displays the most severe change and one additive system that depicts osteochondral and soft tissue-related changes. A total of 47 1.5 T MRI examinations of knees (n = 21) and ankles (n = 26) of 42 haemophilic boys, age range, 22 months to 18 years, performed at different centres (Toronto, n = 20, Europe, n = 12 and Denver, n = 15) were independently reviewed by four radiologists at two occasions. Twenty-two examinations were from children <9 years and 25 from children >/=9. Sagittal and coronal gradient-echo (MPGR, 3D FLASH with fat saturation, GRASS) images were obtained. The MRI examinations of the ankle and knee studies presented with osteochondral abnormalities in 38.5% and 23.8% of the cases respectively. The two scoring systems demonstrated an excellent inter-reader [progressive, 0.88; additive (A, e, s and h components), 0.86] and intra-reader [progressive, 0.92; additive (A, e, s and h components), 0.93] reliability using intraclass correlation coefficients (ICCs). Although ICCs were slightly higher for knees when compared with ankles, and for older children when compared with younger children, all values fell within excellent inter- and intra-reader reliability categories. The two MRI scoring systems demonstrated a comparable reliability. This result constitutes the basis for further development of a combined MRI scoring system for assessment of haemophilic arthropathy, which incorporates progressive and additive components. PMID- 15876271 TI - Cost of severe haemophilia in Toronto. AB - Our objective was to determine costs and trends in treating boys with severe haemophilia A before our centre routinely used prophylaxis. One reviewer extracted data from patient charts to determine resource consumption for 17 boys with severe haemophilia A from 1978 to 1998 at Toronto's Hospital for Sick Children. Resources included factor concentrate, doctors and health care professionals (physiotherapists/social workers), tests (laboratory, radiological and diagnostic) and hospitalizations. Subgroup analysis on those patients infected with HIV and/or hepatitis were also performed. Costs in Canadian Dollars were taken from standard lists and discounted at 3%. Total average cost (range) 62 292 dollars (3339-121 738) per year patient(-1); the largest fraction, 59 910 dollars (3103-119 480), 96.2% was accounted for by factor VIII. Hospitalizations accounted for 1832 dollars (0-5217) per patient year(-1) including drugs, nursing care and stay. Doctor and health care professionals visits averaged 252 dollars (36-462) and 72 dollars (0-175) per patient year(-1), laboratory and other tests cost 201 dollars (22-377) and 26 dollars (2-60) per patient year(-1), respectively. The average number of bleeds was 12.9 (2.0-22.0) per patient year( 1), decreasing since 1977 by 0.68 per patient year(-1) (R(2) = 0.56). Hospitalizations averaged 0.22 (0-4) per patient year(-1), lasting 2.3 days. From 1984, hospitalizations decreased by 0.025 patient(-1) year(-1) (R(2) = 0.76). Concurrently, the average treatment costs increased by 5456 dollars patient(-1) year(-1) (R(2) = 0.81). Clotting factor concentrate cost per patient increased by 5521 dollars year(-1) (R(2) = 0.82). Patients with virally transmitted diseases had considerable higher costs. The cost per year was substantial. Costs increased with virally transmitted diseases. Number of bleeds and hospitalizations over the period of study decreased and costs increased because of factor use in secondary prophylaxis. PMID- 15876272 TI - A cost minimization model for the treatment of minor bleeding episodes in patients with haemophilia A and high-titre inhibitors. AB - Treatment of acute bleeding episodes in patients with haemophilia A and inhibitory antibodies to factor VIII (FVIII) most often involves the use of bypassing haemostatic agents, such as activated prothrombin complex concentrates (aPCC) or recombinant factor VIIa (rFVIIa). We constructed a cost minimization model to compare the costs of initial treatment with aPCC vs. rFVIIa in the home treatment of minor bleeding episodes. We developed a clinical scenario describing such a case and presented it to a panel of US haemophilia specialists. For each product class, we asked panellists to provide dosing regimens required to achieve complete resolution of a minor haemarthrosis in a child with high-titre inhibitors, and for the probabilities of success at two time points (8-12 and 24 h). Consensus among the panellists was refined by a second round of the process, and the median values resulting were used as inputs to a decision analysis model. Sensitivity analyses were conducted to determine threshold values for key variables. The base case model found that initial treatment with aPCC would result in a mean cost per episode of 21 000 dollars, compared with 33 400 dollars for initial treatment with rFVIIa. Sensitivity analyses over a range of clinically plausible values for cost, dosing, and efficacy did not change the selection of aPCC as the dominant strategy. PMID- 15876273 TI - Hepatitis C infection among Dutch haemophilia patients: a nationwide cross sectional study of prevalence and antiviral treatment. AB - Hepatitis C is a major co-morbidity among patients with haemophilia who received inadequately or non-virus-inactivated clotting factor concentrates before 1992. The objectives of this study were to investigate the prevalence of hepatitis C and the use of antiviral therapies during the last decade among patients with haemophilia in the Netherlands. We performed a cross-sectional study and a questionnaire was sent to all 1519 patients known with haemophilia in the Netherlands between 2001 and 2002. The study population for the present study consisted of 771 patients who had received clotting factor products before 1992 of whom 638 reported their hepatitis C status. In total, 441 of the 638 (68%) patients ever had a positive test for hepatitis C virus (HCV); 344 patients (54%) had a current infection, and 97 (15%) had cleared the virus. Among 344 patients currently HCV infected, 111 (32%) had received treatment for hepatitis C, while 34% (33/97) of patients with an infection in the past had been treated for hepatitis C. In 2002 the prevalence of hepatitis C among patients with haemophilia who received clotting factor products before 1992 was 54%. The majority of patients with a current HCV infection had not been treated with antiviral therapy. PMID- 15876274 TI - Pulmonary embolism in a patient with severe congenital deficiency for factor V during treatment with fresh frozen plasma. AB - Thrombosis is a rare complication in patients with congenital clotting factor deficiencies. In most cases, it is related to inherited procoagulant factors, use of central venous catheters or administration of coagulation factor concentrates. There are only a few case reports about thrombotic events during treatment with fresh frozen plasma (FFP). We report the case of a patient with homozygous inherited factor V deficiency, who developed a pulmonary embolism at a time of treatment with methylene blue treated FFP (MBFFP). The patient had only two other factors predisposing to thrombosis and both were acquired: obesity and bed rest. He started anticoagulant treatment with low molecular weight heparin (LMWH) while the deficient factors were replaced with MBFFP. After 8 days of treatment the patient developed a severe respiratory insufficiency. Pulmonary haemorrhage was considered among the differential diagnosis and LMWH was stopped. An inferior vena cava filter was placed without any further thrombotic complications. To our knowledge, there are no reports about patients with clotting factor deficiencies who developed a thrombotic event during treatment with MBFFP. PMID- 15876275 TI - Combined factor V and factor VIII deficiency in a Thai patient: a case report of genotype and phenotype characteristics. AB - A Thai woman, with no family history of bleeding disorders, presented with excessive bleeding after minor trauma and tooth extraction. The screening coagulogram revealed prolonged activated partial thromboplastin time and prothrombin time. The specific-factor assay confirmed the diagnosis of combined factor V and factor VIII deficiency (F5F8D). Her plasma levels of factor V and factor VIII were 10% and 12.5% respectively. The medications and blood product treatment to prevent bleeding from invasive procedure included 1-deamino-8-d arginine vasopressin, cryoprecipitate, factor VIII concentrate, fresh frozen plasma and antifibrinolytic agent. Gene analysis of the proband identified two LMAN1 gene mutations; one of which is 823-1 G --> C, a novel splice acceptor site mutation that is inherited from her father, the other is 1366 C --> T, a nonsense mutation that is inherited from her mother. Thus, the compound heterozygote of these two mutations in LMAN1 cause combined F5F8D. PMID- 15876276 TI - Successful steroid pulse treatment in childhood acquired haemophilia with nephrotic syndrome. AB - We encountered a 2-year-old boy with acquired haemophilia, which rarely occurs in children, who was complicated with nephrotic syndrome. In mid-August 2001, he was diagnosed to have nephrotic syndrome based on the presence of massive proteinuria and hypoalbuminaemia. Activated partial thromboplastin time (APTT) was normal at 42.4 s at that time. After starting prednisone administration of 2 mg kg(-1) day( 1), the proteinuria disappeared immediately. However, in early October the same year, subcutaneous ecchymosis and intramuscular bleeding occurred for no apparent reason, and from the examination results his APTT was 106.4 s, factor VIII (FVIII) activity was <1%, and the anti-FVIII inhibitor titre was 6.9 BU ml(-1). As a result, he was diagnosed to have acquired haemophilia. The anti-nuclear antibody and anti-phospholipid antibody were negative. With recombinant activated FVII, haemostasis was obtained, and after administering three courses of steroid pulse therapy (methyl prednisolone: 20 mg kg(-1) day(-1) x 3 days), the anti FVIII inhibitory activity disappeared. An analysis of the immunological and coagulation properties of his FVIII autoantibodies showed the anti-FVIII inhibitory activity to be mediated by IgG(1) antibody. In other words, his FVIII inhibitor was a Th1 dominant and it provided a good response to treatment. These findings correlate with those of previous reports. The patient thereafter frequently demonstrated a recurrence of nephrotic syndrome. As a result, he is presently being managed with cyclosporine. However, no recurrence of the anti FVIII titre has been observed. PMID- 15876277 TI - Knocking down the price of factor concentrates in Brazil. PMID- 15876278 TI - The safety and efficacy of B-domain deleted recombinant factor VIII concentrates in patients with severe haemophilia A: an update. PMID- 15876279 TI - Osteoprotegerin and bone turnover markers in heavily pretreated HIV-infected patients. AB - OBJECTIVES: To characterize osteoprotegerin (OPG) levels, bone remodelling and bone mineral density (BMD) in heavily pretreated HIV-infected patients on antiretroviral therapy, and to evaluate the clinical factors associated with bone density decline. METHODS: Heavily pretreated (> 5 years) HIV-positive patients were enrolled in this cross-sectional, observational study, which was based on a total body bone densitometry examination and a comprehensive evaluation of bone and mineral parameters. RESULTS: Sixty-eight patients (55 male and 13 female) with a median age of 41 years (range 25-60 years) were included in the study. Their antiretroviral treatment lasted for 82 months. On the basis of the World Health Organization criteria, nine patients (13.2%) were osteoporotic [T-score < 2.5 standard deviation (SD)] and 19 patients (27.9%) were osteopenic (T-score between -1 and -2.5). The principal outcomes associated with the presence of a low BMD were high OPG and lysylpyridinoline/creatinine ratio (Dpd) values. Most of the patients (39 of 48; 81.25%) showed vitamin D insufficiency [Vitamin D (25(OH)D) < 18 ng/mL] with secondary hyperparathyroidism (13 of 50 patients: 26%), which proved to be correlated to osteocalcin (BGP) levels [parathyroid hormone (PTH) vs. BGP: r = 0.34; P < 0.01]. There was an inverse correlation between T-scores and serum osteocalcin and alkaline phosphatase (AP) levels, on one hand, and Dpd, on the other. High AP and Dpd values were associated with relative risks of 4.1 [95% confidence interval (CI) = 1.01-17.6] and 7.2 (95% CI = 1.67-31.03), respectively, of a pathological T-score. Multivariate analysis revealed that the factors associated with the presence of osteopenia or osteoporosis were older age and lower body mass index. CONCLUSIONS: About 40% of our heavily pretreated subjects with advanced HIV infection had a low BMD, and 56% (24 of 44 patients) showed a high bone turnover rate with marked osteoclast activation. High OPG levels may protect against bone resorption. PMID- 15876280 TI - The durability of virological success of tenofovir and didanosine dosed at either 400 or 250 mg once daily. AB - Tenofovir (TDF) co-administered with didanosine (ddI) 400 mg increases ddI plasma concentrations by up to 60%, raising concerns over toxicity. To limit this interaction, the dosage of ddI may be reduced to 250 mg once daily when co prescribed with TDF. In this clinical cohort, highly active antiretroviral therapy regimens containing TDF and ddI 250 mg were significantly better tolerated than combinations with TDF and ddI at a dose of 400 mg. Low-dose ddI 250 mg once daily plus TDF as part of antiretroviral therapy was effective. PMID- 15876281 TI - Antiretroviral treatment change among HIV, hepatitis B virus and hepatitis C virus co-infected patients in the Australian HIV Observational Database. AB - OBJECTIVES: To assess the impact of highly active antiretroviral therapy (HAART) on rates of change of antiretroviral treatment among patients co-infected with hepatitis B virus (HBV) and/or hepatitis C virus (HCV) in the Australian HIV Observational Database (AHOD). METHODS: Analysis was based on 805 of the 2218 patients recruited to the AHOD by March 2003, who had commenced HAART after 1 January 1997, who had recorded test results for HBV surface antigen and anti-HCV antibody, and who had follow-up of more than 3 months. The effect of hepatitis co infection on the rate of antiretroviral treatment change after commencing HAART was assessed using a random-effect Poisson regression model. RESULTS: Among those included in the analyses, the prevalences of HBV and HCV were 4.8% and 12.8%, respectively. The overall rate of combination antiretroviral treatment change was 0.74 combinations per year. Factors independently associated with an increased rate of change of combination antiretroviral treatment were: prior AIDS-defining illness; prior exposure to double combination antiretroviral therapy; and antiretroviral treatment class. Co-infection with HBV and/or HCV was not found to be significantly associated with the rate of combination antiretroviral treatment change. CONCLUSIONS: While both HBV and HCV co-infections are relatively common in the AHOD, they do not appear to be serious impediments to the treatment of HIV infected patients. PMID- 15876282 TI - Effects of antiretroviral treatment on blood-brain barrier integrity and intrathecal immunoglobulin production in neuroasymptomatic HIV-1-infected patients. AB - OBJECTIVES: To study the effect of antiretroviral combination therapy on blood brain barrier (BBB) integrity and intrathecal immunoglobulin G (IgG) production. METHODS: Lumbar punctures were performed on 38 neurologically asymptomatic, treatment-naive HIV-1-infected patients prior to and during treatment at intervals of approximately 4 months, 1 year and 2 years. Albumin ratio and IgG index were analysed as markers of BBB integrity and intrathecal IgG synthesis. RESULTS: HIV-1 RNA decreased to < 50 HIV-1 RNA copies/mL in the cerebrospinal fluid (CSF) of all patients and in the plasma of all but one patient. Only 5% of patients had elevated albumin ratio values at baseline, while 56% had an elevated IgG index. There was no significant reduction of the albumin ratio or the IgG index. After 2 years of treatment all patients had normal albumin ratio values, while 41% still had increased IgG index levels. CONCLUSIONS: Up to 2 years after the initiation of treatment, the favourable impact of antiretroviral combination treatment on CSF viral load was not accompanied by a similar reduction of intrathecal IgG production. BBB function, measured as the albumin ratio, was not significantly changed in this cohort of neurologically asymptomatic HIV-1 infected patients. PMID- 15876283 TI - Gender difference in HIV-1 RNA viral loads. AB - OBJECTIVES: To test and characterize the dependence of viral load on gender in different countries and racial groups as a function of CD4 T-cell count. METHODS: Plasma viral load data were analysed for > 30,000 HIV-infected patients attending clinics in the USA [HIV Insight (Cerner Corporation, Vienna, VA, USA) and Plum Data Mining LLC (East Meadow, NY, USA) databases] and the Netherlands (Athena database; HIV Monitoring Foundation, Amsterdam, Netherlands). Log-normal regression models were used to test for an effect of gender on viral load while adjusting for covariates and allowing the effect to depend on CD4 T-cell count. Sensitivity analyses were performed to test the robustness of conclusions to assumptions regarding viral loads below the lower limit of quantification (LLOQ). RESULTS: After adjusting for covariates, women had (nonsignificantly) lower viral loads than men (HIV Insight: -0.053 log(10) HIV-1 RNA copies/mL, P = 0.202; Athena: -0.005 log(10) copies/mL, P = 0.667; Plum: -0.072 log(10) copies/mL, P = 0.273). However, further investigation revealed that the gender effect depended on CD4 T-cell count. Women had consistently higher viral loads than men when CD4 T-cell counts were at most 50 cells/microL, and consistently lower viral loads than men when CD4 T-cell counts were greater than 350 cells/microL. These effects were remarkably consistent when estimated independently for the racial groups with sufficient data available in the HIV Insight and Plum databases. CONCLUSIONS: The consistent relationship between gender-related differences in viral load and CD4 T-cell count demonstrated here explains the diverse findings previously published. PMID- 15876284 TI - The Protease Inhibitor Transfer Study (PROTRA 1): abacavir and efavirenz in combination as a substitute for a protease inhibitor in heavily pretreated HIV-1 infected patients with undetectable plasma viral load. AB - OBJECTIVES: The aim of the study was to evaluate the safety and efficacy of abacavir (ABC) and efavirenz (EFV) instead of a protease inhibitor (PI) in HIV-1 infected subjects treated with two nucleoside reverse transcriptase inhibitors (NRTIs) and one PI with undetectable viral loads (< 50 HIV -1 RNA copies/mL). To be eligible for inclusion, patients had to have a history of viral load < 400 copies/mL for at least 3 months and had to be naive to treatment with nonnucleoside reverse transcriptase inhibitors (NNRTIs) and ABC, but multiple pretreatment and treatment failure were allowed. DESIGN: An open-label, single centre pilot study of duration 48 weeks was conducted. ABC was added to the original treatment with two NRTIs and one PI at baseline, and at week 6 the PI was replaced by EFV. At each study visit, CD4 cell count, viral load [measured by polymerase chain reaction (PCR)] and clinical chemistry were measured. Fasting blood samples were taken at baseline and at weeks 12, 24, 36 and 48 to measure levels of cholesterol [high-density lipoprotein (HDL)/low-density lipoprotein (LDL)], triglycerides, insulin and C-peptide. Additionally, an oral glucose tolerance test (OGTT) was performed. A bioelectric impedance analysis (BIA) and a single slice abdominal and mid-thigh computed tomography (CT) scan were carried out to assess changes in body composition. RESULTS: Thirty patients were included in the study. Three patients experienced ABC-hypersensitivity and one patient demonstrated virological failure caused by nonadherence. At week 48, all remaining patients had viral loads < 50 copies/mL with stable CD4 counts. The fasting metabolic parameters and abdominal fat distribution remained unchanged. CONCLUSIONS: In heavily pretreated patients, ABC and EFV in combination provide an effective, simplified and well-tolerated alternative to PI treatment. PMID- 15876285 TI - Better maintained adherence on switching from twice-daily to once-daily therapy for HIV: a 24-week randomized trial of treatment simplification using stavudine prolonged-release capsules. AB - BACKGROUND: Adherence to antiretroviral therapy is critical to treatment outcomes. Adherence studies in other therapeutic areas of medicine suggest that once-daily regimens support improved adherence when compared to twice-daily therapy. An expansion in the range of once-daily antiretrovirals is making once daily therapy possible for persons with HIV infection. METHODS: A 24-week randomized open-label simplification study of twice-daily regimens based on stavudine immediate release or zidovudine to an all once-daily regimen based on the stavudine prolonged-release capsule (PRC), in persons with complete virological suppression on regimens also including efavirenz and lamivudine, was carried out. Subjects were assessed for adherence [using the Medication Event Monitoring System (MEMS) cap; Aardex Corporation, Union City, CA, USA], quality of life, tolerability and efficacy. RESULTS: Forty-three patients were randomly assigned: 21 remained on their original regimen and 22 switched to once-daily therapy with stavudine PRC. Although high levels of adherence and good quality of life were present at study enrollment, adherence declined to a significantly lesser extent at week 24 in the group that switched to once-daily therapy. Efficacy was maintained in both groups and there were no differences in tolerability or toxicity. CONCLUSIONS: Subjects switching from twice-daily therapy to once-daily therapy demonstrate less of a decline in adherence over 24 weeks. A once-daily regimen including stavudine PRC is as effective and tolerable as a regimen containing the twice-daily formulation. PMID- 15876286 TI - The use of a triple nucleoside-nucleotide regimen for nonoccupational HIV post exposure prophylaxis. AB - OBJECTIVES: Nonoccupational post-exposure prophylaxis (NPEP) for HIV is recommended after high-risk sexual exposure. Because of the high incidence of intolerable side effects observed with protease inhibitor- and zidovudine-based NPEP regimens, our unit changed standard NPEP treatment to 28 days of tenofovir lamivudine-stavudine (TDF-3TC-d4T). The aim of this study was to compare side effects and numbers of individuals completing NPEP before and after this change. METHODS: Parameters were compared amongst individuals commencing the following NPEP regimens: zidovudine-lamivudine (ZDV-3TC), zidovudine-lamivudine-nelfinavir (ZDV-3TC-NFV) and TDF-3TC-d4T. RESULTS: A total of 385 individuals received ZDV 3TC (n = 36), ZDV-3TC-NFV (n = 225) or TDF-3TC-d4T (n = 137) as NPEP for the first time between June 1999 and November 2003. Noncompletion rates were 25%, 32% and 15%, respectively (P = 0.001), with odds ratios for noncompletion being 2.0 [95% confidence interval (CI) 0.8-4.8] and 2.7 (95% CI 1.6-4.8) in the first two groups compared with the TDF-3TC-d4T group (P = 0.008). Adverse events were less common in the TDF-3TC-d4T group, with significantly lower rates of nausea and headache, but significantly higher rates of peripheral neuropathy and asymptomatic raised transaminases. There was no HIV seroconversion in any group. CONCLUSIONS: TDF-3TC-d4T is significantly better tolerated than ZDV-3TC or ZDV 3TC-NFV as NPEP and results in greater numbers of individuals completing 28 days of treatment. PMID- 15876287 TI - Determinants of clinical progression in antiretroviral-naive HIV-infected patients starting highly active antiretroviral therapy. Aquitaine Cohort, France, 1996-2002. AB - OBJECTIVES: To determine the factors associated with clinical progression (AIDS events and death) in antiretroviral-naive patients who have begun highly active antiretroviral therapy (HAART). METHODS: HIV-infected patients naive to antiretroviral therapy were included in a prospective hospital-based cohort who began HAART between June 1996 and December 2001. Progression was explained by baseline characteristics using Cox proportional hazards models. RESULTS: Overall, data for 709 patients were analysed. In multivariate analysis, factors associated with an increased risk of progression were CD4 count < 50 cells/microL [hazard ratio (HR) = 13.0 (95% confidence interval 3.8-44.3)] and between 50 and 199 cells/microL [HR = 5.1 (1.6-16.3)], when compared with patients with CD4 count>350 cells/microL; AIDS events before HAART prescription [HR = 2.1 (1.2 3.7)]; CD8 count < 400 cells/microL [HR = 1.8 (1.1-3.0)]; and older age (HR = 1.2 by 10 years (1.0-1.5)]. In a second model including CD4 percentage, factors associated with progression were CD4 < 10% [HR = 6.3 (2.2-17.9)] and 10% 20%; CD8 count; AIDS events before HAART prescription; and older age. In a third model including the CD4:CD8 ratio, factors associated with progression were CD4:CD8 < 15% [HR = 8.2 (2.3-28.8)] and 15% < CD4:CD8 < 30% [HR = 4.6 (1.3-16.0)], when compared with patients with CD4:CD8 > 45%; AIDS events before HAART prescription; and older age. The Akaike information criteria for model analysis were 803, 805 and 815, respectively. CONCLUSIONS: Consideration of CD4 level in terms of CD4:CD8 ratio or CD4 percentage can be a good alternative to absolute CD4 count. Other prognostic factors such as older age, CD8 count < 400 cells/microL and AIDS events also have to be considered in the decision to initiate HAART. PMID- 15876288 TI - Prevalence of hepatitis C in an ethnically diverse HIV-1-infected cohort in south London. AB - OBJECTIVES: There is limited information on the prevalence of and risk factors for hepatitis C virus (HCV) infection among HIV-1-infected patients in the UK. Our objective was to determine the prevalence of HCV infection among an ethnically diverse cohort of HIV-infected patients in south London, and to extrapolate from these data the number of co-infected patients in the UK. METHODS: A total of 1017 HIV-1-infected patients who had attended King's College Hospital HIV clinic between September 2000 and August 2002 were screened for HCV antibody using a commercial enzyme-linked immunosorbent assay (ELISA). Positive results were confirmed by polymerase chain reaction (PCR) or recombinant immunoblot assay. Demographic, clinical and laboratory data were obtained from the local computerized database and medical records. We applied our HCV prevalence rates in the different HIV transmission groups to the estimated number of HIV-infected persons in these groups in the UK, to obtain a national estimate of the level of HIV-HCV co-infection. RESULTS: Of the 1017 HIV-1-infected patients, 407 (40%) were white men, 158 (15.5%) were black African men, 268 (26.3%) were black African women, and 61 (6%) and 26 (2.6%) were black Caribbean men and women, respectively. Heterosexual exposure was the most common route of HIV acquisition (53.5%), followed by men having sex with men (36.9%), and current or previous injecting drug use (IDU) (7.2%). The overall prevalence of HCV co infection was 90/1017 (8.9%), but this varied substantially according to route of transmission, from 82.2% among those with a history of IDU (which accounted for 67% of all HCV infections), to 31.8% in those who had received blood products, to 3.5% and 1.8% in those with homosexually and heterosexually acquired infection, respectively. Multivariate logistic regression analysis identified several independent risk factors for HCV infection: a history of IDU [odds ratio (OR) = 107.2; 95% confidence interval (CI) = 38.5-298.4], having received blood products (OR = 16.5; 95% CI = 5.1-53.7), and either being from a white ethnic group (OR = 4.3; 95% CI = 1.5-12.0) or being born in Southern Europe (OR = 6.7; 95% CI = 1.5 30.7). Based on the 35,473 known HIV-1-infected persons in the UK and the 10 997 estimated to be unaware of their status, we projected that there are at least 4136 HIV-HCV co-infected individuals in the UK and 979 who are unaware of their status. CONCLUSIONS: Overall, 9% of our cohort was HIV-HCV co-infected. The prevalence was highest among intravenous drug users (82%), who accounted for most of our HCV cases, and lowest among heterosexual men and women from sub-Saharan Africa and the Caribbean [< 2%]. Our estimate that a significant number of co infected persons may be unaware of their HIV and HCV status, highlights an urgent need to increase the uptake of HCV and HIV testing, particularly among injecting drug users, to reduce the risk of onward transmission. PMID- 15876289 TI - Predicting short-term disease progression among HIV-infected patients in Asia and the Pacific region: preliminary results from the TREAT Asia HIV Observational Database (TAHOD). AB - OBJECTIVES: HIV disease progression has been well documented in Western populations. This study aimed to estimate the short-term risk of AIDS and death from the TREAT Asia HIV Observational Database (TAHOD), a prospective, multicentre cohort study in Asia and the Pacific region. METHODS: Prospective data were analysed to estimate short-term disease progression. Endpoints were defined as the time from study entry to diagnosis with AIDS or death. Antiretroviral treatment was fitted as a time-dependent variable. Predictors of disease progression were assessed using Cox proportional hazards models, and prognostic models were developed using Weibull models. RESULTS: A total of 1260 patients with prospective follow-up data contributed 477 person-years of follow up, during which 18 patients died and 34 were diagnosed with AIDS, a combined rate of 10.1 per 100 person-years. Compared with patients receiving antiretroviral treatment, patients not on treatment had a higher rate of disease progression (17.6 vs. 8.1 per 100 person-years, respectively). Baseline CD4 count was the strongest predictor of disease progression. Prognostic models, using either a baseline CD4 count as the sole marker or markers including baseline haemoglobin, AIDS-related symptoms and previous or current antiretroviral treatment, were successful at identifying patients at high risk of short-term disease progression. CONCLUSIONS: Similar to the situation in Western countries, baseline CD4 count was the strongest predictor of short-term disease progression. Prognostic models based on readily available clinical data and haemoglobin level should be useful in estimating short-term clinical risk in HIV-infected patients in Asia and the Pacific region. PMID- 15876290 TI - A comparative analysis of periapical health based on historic and current data. AB - AIM: To compare periapical health using samples from prehistoric and historic periods until the present day, and to emphasize the major risk indicators for apical periodontitis (AP). METHODOLOGY: A comparative survey to assess periapical health was performed on five samples (525 individuals) drawn from different periods of history within the time frame 2000 BC to 2000 AD. Twenty-one binary risk indicators for AP were retained for a logistic regression model. The probability of a diseased tooth was defined from a two-level response variable based on the periapical index (PAI). An individual regression model was computed with partial least squares (PLS) regression model, based on the individual mean values of the nine retained risk indicators. RESULTS: Condensing osteitis, tooth wear, caries, root fillings and the presence of inadequate root fillings were associated with the PAI levels. The maxillary molars and recent time periods (contemporaneous and seventeenth century) were also risk indicators for the pathological condition. The PLS regression for individuals demonstrated correlations between risk factors. This multidimensional analysis indicated that the mean PAI was correlated mainly with caries and condensing osteitis. Condensing osteitis was more frequent in the mandibular than in the maxillary bone (P=0.001), and correlated with tooth wear in ancient periods. CONCLUSIONS: This comparative analysis demonstrated a significantly higher prevalence of AP in the contemporary period. The most important risk indicators for that period were the presence of inadequate root fillings, carious lesions and condensing osteitis. PMID- 15876291 TI - The effect of application time of EDTA and NaOCl on intracanal smear layer removal: an SEM analysis. AB - AIM: To verify, under the scanning electron microscope (SEM), the influence of irrigation time with ethylenediaminetetraacetic acid (EDTA) and sodium hypochlorite (NaOCl) on intracanal smear layer removal. METHODOLOGY: Twenty-one extracted human permanent teeth with single straight root canals were included. The root canals of the teeth were instrumented and, at the end of preparation, were irrigated with 3 mL of 15% EDTA, followed by 3 mL of 1% NaOCl for 1 min (group 1), for 3 min (group 2), and for 5 min (group 3). The canals of teeth in group 4 (control) did not receive the final irrigation. The teeth were sectioned longitudinally and prepared for an SEM. The dentinal wall of cervical, middle and apical thirds was graded according to the amount of debris and smear layer remaining on the walls. The results were analysed using the Kruskal-Wallis and Conover-Inman tests. RESULTS: In all the canals of experimental groups irrigation with EDTA and NaOCl completely removed the smear layer from the cervical and middle thirds. In the apical third, the dentine surface were partially covered, particularly in the teeth of group 1, where there was significantly more smear layer when compared with the other thirds in the same group (P<0.007). However, the Kruskal-Wallis test showed overall that there were no significant differences between groups 1, 2 and 3 (P>0.05). CONCLUSION: In this limited laboratory study, canal irrigation with EDTA and NaOCl for 1, 3 and 5 min were equally effective in removing the smear layer from the canal walls of straight roots. PMID- 15876292 TI - Pressures generated in vitro during Stabident intraosseous injections. AB - AIM: To test the hypothesis that the Stabident intraosseous injection is a potentially high-pressure technique, which carries serious risks of anaesthetic cartridge failure. METHODOLOGY: A standard Astra dental syringe was modified to measure the internal pressure of local anaesthetic cartridges during injection. Intra-cartridge pressures were measured at 1 s intervals during slow (approximately 15 s) and rapid (<10 s) injections of 2% Xylocaine with 1:80,000 adrenaline (0.25 cartridge volumes) into air (no tissue resistance), or into freshly prepared Stabident perforation sites in the anterior mandible of freshly culled young and old sheep (against tissue resistance). Each injection was repeated 10 times over 3 days. Absolute maximum pressures generated by each category of injection, mean pressures at 1 s intervals in each series of injections, and standard deviations were calculated. Curves of mean maximum intra cartridge pressure development with time were plotted for slow and rapid injections, and one-way anova (P<0.05) conducted to determine significant differences between categories of injection. RESULTS: Pressures created when injecting into air were less than those needed to inject into tissue (P<0.001). Fast injection produced greater intra-cartridge pressures than slow delivery (P<0.05). Injection pressures rose more quickly and to higher levels in small, young sheep mandibles than in larger, old sheep mandibles. The absolute maximum intra-cartridge pressure developed during the study was 3.31 MPa which is less than that needed to fracture glass cartridges. CONCLUSIONS: Stabident intraosseous injection conducted in accordance with the manufacturer's instructions does not present a serious risk of dangerous pressure build-up in local anaesthetic cartridges. PMID- 15876293 TI - Collagenase-3 (MMP-13) is expressed in periapical lesions: an immunohistochemical study. AB - AIM: To determine whether or not matrix metalloproteinase 13 (MMP-13) is present in periapical granulomas with and without epithelium. METHODOLOGY: Seventeen open periapical granulomas of pulpal origin (seven lesions without epithelium and 10 with proliferating epithelium) were fixed in formalin and then embedded in paraffin prior to being processed for immunohistochemical analysis. A monoclonal antibody against human MMP-13 was used to evaluate MMP-13 expression. Immunocomplexes were subsequently treated with the secondary antibody and then detected by means of streptavidin peroxidase. Immunoreactivity was visualized by development with 3,3'-diaminobenzidine. RESULTS: An immunopositive cytoplasmatic reaction for MMP-13 was observed in all the specimens, although the immunostaining by anti-MMP-13 antibody was heterogeneous and its levels varied according to histopathological findings. In periapical lesions without epithelium MMP-13 immunolabelling was detected in a few fibroblast-like cells, and in some plasma cells within the granulomatous tissue. A clear upregulation of MMP-13 expression was detected in periapical lesions with epithelium, especially in small island and thin strands of epithelium. CONCLUSIONS: The expression pattern of MMP-13 demonstrates that it is involved in the conversion of a periapical granuloma with epithelium into a radicular cyst. This property is related to the ability of MMP-13 to influence not only the migration of epithelial cell but also the invasion of granulomatous tissue. PMID- 15876294 TI - The sealing ability of an epoxy resin root canal sealer after Nd:YAG laser irradiation of the root canal. AB - AIM: To evaluate ex vivo the effect of Nd:YAG laser irradiation with and without black ink on instrumented root canal walls, and the degree of both coronal and apical microleakage of filled root canals. METHODOLOGY: Seventy-two single-rooted teeth were instrumented up to a size 40 K-file, and then divided into six groups of 10 teeth: groups 1 and 4 remained unlased and acted as control groups, groups 2 and 5 were treated with a Nd:YAG laser (Fidelis Plus, Herzele, Belgium), groups 3 and 6 were treated with a laser and black ink; the remaining 12 teeth served as positive and negative controls. The laser was operated at 1.5 W, 15 Hz, four times for 5 s with a 20-s interval. Groups 4-6 were filled using cold lateral condensation of gutta-percha and AH26. After storage in water for 48 h at 37 degrees C, through-and-through leakage (L in microL day-1) was measured for 48 h under a pressure of 1.2 atm using a fluid transport model and recorded as L=0 (L1), 010 (L3). After the assessment of leakage with the fluid transport model, the teeth were immersed in rhodamine B solution for 48 h at 37 degrees C. Apical and coronal dye leakage was scored after longitudinal splitting of these teeth. All teeth of groups 1-3 were split longitudinally and observed under SEM for evaluation of remaining smear layer. RESULTS: Through-and-through leakage was only observed in the group lased with black ink (two samples-L2). Apical and coronal dye leakage was observed in all groups; there were no statistically significant differences amongst the three experimental groups. The through-and-through leakage, measured with the fluid transport model in two teeth of group 6, was confirmed in the dye leakage test (rhodamine B dye was observed along the total length of the root filling). There was evidence of melted and ablated root canal dentine in the laser-treated groups. These findings were more obvious in root canals lased in association with black ink. All apical foramina in the lased group remained patent. CONCLUSIONS: Nd:YAG laser irradiation with black ink increased the amount of melted and ablated dentine areas compared with that without black ink. Nd:YAG lasing in association with black ink did not result in a reduction of either coronal or apical microleakage in root filled teeth. PMID- 15876295 TI - SEM analysis of the integrity of resected root apices of cadaver and extracted teeth after ultrasonic root-end preparation at different intensities. AB - AIM: To compare the integrity of root apices of cadaver and extracted teeth after resection, ultrasonic root-end cavity preparation at medium and low ultrasonic power settings and retrieval. METHODOLOGY: Root canal treatment, perpendicular root-end resection and root-end preparation were performed on single-rooted anterior and premolar teeth (49 teeth in situ in maxillary and mandibular jaws from cadavers and 45 extracted teeth). Apical root-end cavities were prepared with the S12/90 degrees D tip and the Suni-Max ultrasonic unit (Satelec, Merignac, France) at the intensity prescribed by the manufacturer (power 7 at power mode S) (34 cadaver teeth, 30 extracted teeth) and at a lower intensity (power 4 at power mode S) (15 cadaver teeth, 15 extracted teeth). After ultrasonic preparation the cadaver teeth were retrieved from the jaws. Exaflex impressions (GC Corporation, Tokyo, Japan) were made of the root apices after resection, root-end preparation and retrieval. These impressions were processed for SEM analysis, and the recordings evaluated for cracks and marginal chipping. RESULTS: In general, extracted teeth showed significantly more cracks and chipping than cadaver teeth. Lowering the ultrasonic power from medium to low intensity resulted in equal scores for cracks on extracted teeth and for chipping on cadaver teeth, in higher scores for cracks on cadavers and in lower scores for chipping on extracted teeth. Complete cracks and cracks originating from the root surface occurred only in extracted teeth. CONCLUSIONS: The number of cracks and degree of chipping caused by ultrasonic root-end preparation was higher on extracted teeth than on cadaver teeth. Lowering the ultrasonic power from medium to low intensity cannot be recommended as it resulted in more cracks and equal chipping on cadaver teeth. Investigation of techniques and materials should be conducted in situ and not on extracted teeth. PMID- 15876296 TI - Periapical resurgery versus periapical surgery: a 5-year longitudinal comparison. AB - AIM: To monitor and compare the outcome of periradicular surgery in teeth that had previously undergone surgical treatment versus teeth that were undergoing a surgical procedure for the first time. METHODOLOGY: A total of 164 patients with 231 roots with previously unresolved periapical lesions were followed for up to 5 years following surgery. In all, 162 roots had received unsuccessful conventional root canal treatment (group AS) and 69 had been treated previously using apical surgery (group RS). The surgical procedure was completed using ultrasonic retrotips to prepare the apical root-end cavity, and a zinc-oxide EBA reinforced material was used to fill the apical root-end cavities. Lesions were radiologically examined from 1 to 5 years following the surgical procedure. Radiographs were independently analysed, according to a previously published classification. RESULTS: In all the roots examined after 5 years, the overall healing rate was 78%; in group AS, 140 (86%) healed with complete bone filling of the surgical cavity, 12 (7%) were considered to have incompletely healed and 10 (6%) were associated with post-treatment disease. In group RS, 41 (59%) healed completely, 12 (17%) were considered to have incompletely healed and 16 (23%) were associated with post-treatment disease. The difference between a second surgical procedure and a first surgical procedure was statistically significant (Mann-Whitney U-test, P=0.001). CONCLUSION: Surgical retreatment of teeth previously treated with surgery is a valid alternative to extraction. However, association with post-treatment disease was greater than after a primary surgical approach. PMID- 15876297 TI - Upregulation of tissue-type plasminogen activator in inflamed human dental pulps. AB - AIM: To compare tissue-type plasminogen activator (t-PA) expression in normal human pulp and inflamed human pulp tissue specimens. METHODOLOGY: Thirty pulpal tissue specimens (13 normal and 17 inflamed pulps) were obtained from extracted third molars. The levels of t-PA between normal pulp and inflamed pulp tissues were compared using the quantitative reverse-transcriptase polymerase chain reaction analysis. In addition, immunohistochemistry was used to identify the in situ localization of t-PA expression in pulp specimens. Wilcoxon-Mann-Whitney rank sum test was applied for the statistical analysis of the results. RESULTS: t PA mRNA gene was found more in inflamed pulps when compared with normal pulp tissue (P<0.05). The results from immunohistochemistry demonstrated that t-PA expression was significantly higher in the inflamed pulp (P=0.025). t-PA stain was detected in the fibroblasts, inflammatory infiltrates and endothelial cells. CONCLUSIONS: t-PA expression was significantly higher in inflamed pulp tissue. t PA may play an important role in the pathogenesis of pulpal inflammation. PMID- 15876298 TI - Unicystic ameloblastoma: a possible pitfall in periapical diagnosis. AB - AIM: To report a clinical case of unicystic ameloblastoma previously misdiagnosed as radicular cyst. SUMMARY: A 49-year-old white male was referred to a private practitioner complaining of an asymptomatic bony hard swelling of the left posterior mandible. The patient's dental history indicated that his left mandibular first molar had been extracted approximately 10 years previously. At that time, preoperative radiographic examination demonstrated a radiolucent area of 1.5 cm diameter with well-defined margins involving the distal root of tooth 36. The lesion was diagnosed as cystic and surgery for its removal was advised, but not performed. At presentation, radiography demonstrated a well-defined 3 cm diameter radiolucency extending from the second premolar to the second molar. The lesion was enucleated and histopathological examination confirmed a diagnosis of unicystic ameloblastoma. KEY LEARNING POINTS: *Despite a clinical diagnosis of periapical disease of endodontic origin, a nonendodontic lesion may be present. *Unicystic ameloblastoma located on the periapical area of a tooth can lead to a pulp-periapical misdiagnosis, and should be considered in differential diagnosis. *All tissue specimens recovered in apical surgery should be submitted to histopathological analysis. PMID- 15876300 TI - The respiratory system in connective tissue disorders. AB - The connective tissue disorders (also called collagen vascular diseases) represent an heterogeneous group of immunologically mediated inflammatory disorders with a large variety of affected organs besides the lungs. The respiratory system may be involved in all its components: airways, vessels, parenchyma, pleura, respiratory muscles, etc. The frequency, clinical presentation, prognosis and response to therapy vary, depending on the pattern of involvement as well as on the underlying connective tissue disorders. The subject of this review is to describe the most frequent type of lung disorders observed in patients with connective tissue disease (CTD). We will focus on the most frequent CTD: systemic lupus erythematosus, rheumatoid arthritis, scleroderma, Sjogren's syndrome, dermatopolymyositis and mixed CTD. PMID- 15876301 TI - How can we improve compliance in pediatric pneumology and allergology? AB - Not too long ago, physicians took 'compliance' or 'adherence' to mean the strict obedience of patients and parents - doing what the physician said, suggesting that the success or failure of the treatment lay entirely in the hands of the patient. But it is preferable to improve compliance rather than enforce it. As there is no single solution that will ensure compliance, several steps should be taken: (i) 'exploration' includes open questions, and allows admission of non compliance, e.g. in terms of failure to take drugs, or smoking; (ii) 'education' means explaining pathomechanisms, and reducing corticophobia; (iii) 'tailoring' is reached by focussing therapeutic options on individual requirements, and simplifying treatment regimens; (iv) 'contracting' involves negotiating a realistic therapeutic regimen on an individual basis; (v) 'reminders' are practical tips, which may help to overcome reluctance, e.g. by placing drugs in the toothbrush glass, and on the breakfast table, or in the sports bag; (vi) 'follow-up' ensures long-term compliance, with regular visits to the physician, and self-management programmes. In conclusion, non-compliance of patients and parents is a challenging problem in daily practice. Including the aspects of the abovementioned factors can help build up a relation of trust and partnership, which is the prerequisite of any therapeutic success. However, this is an ongoing process, which has to be continuously reviewed. PMID- 15876302 TI - Continuing Medical Education: an international reality. AB - We are all aware today of the growing interest in continuing medical education (CME) programmes in many European Countries and it is important to understand why and how CME could become an international reality. It is obvious that patients need a good doctor--the best possible--as far as medical knowledge, attention to the patient's quality of life and cost-control is concerned. All European health care systems have to take into consideration everything that causes patient dissatisfaction, risk management and unjustified expenses. An example is the increase of claims and complaints against doctors and the strong attention of patients to medical procedures. In other words, medicine worldwide is becoming a service industry and has to consider quality and quantity of performances as well as to pay attention to personal responsibility. The object of our work is to evaluate the CME systems present in Europe, to show the work done on CME by the CME Committee of the European Academy of Allergology and Clinical Immunology and to highlight the Consensus Report on CME approved by an international panel of CME experts. PMID- 15876303 TI - In search for predictive factors for atopy in human cord blood. AB - Since early prevention is regarded as an important corner stone in the management of atopic diseases, the identification of reliable markers detecting individuals at risk are of major interest. Therefore, many efforts have been made to unravel reliable predictors for atopy which might identify children at risk and allow the initiation of preventive strategies at an early stage. In the past, much scientific energy has been forced in particular on the development of as noninvasive methods as possible to reach this goal. It is obvious that the identification of markers for atopy at the earliest time of life - namely immediately after birth - represents one of the most attractive attempts. In consequence various studies have been initiated to address this issue investigating markers for atopy in cord blood. Most of them have been geared to our current knowledge about cellular and soluble factors which are dysregulated in adolescent atopic individuals. Although the findings of these studies will improve our knowledge about the initial evolution of atopy, several parameters evaluated did not show any association or have led to almost conflicting results. In order to provide an up-date about the current developments in this field, recent research findings on predictive factors for atopy in cord blood are summarized in the following synopsis. PMID- 15876304 TI - Evaluation of an intranasal house dust mite provocation model as a tool in clinical research. AB - BACKGROUND: As a result of the all-year-round exposure to house dust mite (HDM), perennial rhinitis patients never have a clear symptom-free period. In this study, we investigated whether, despite these symptoms, we can still use nasal HDM provocations to study perennial allergic rhinitis and the effects of treatment. METHODS: In a parallel-group study, after 1 week treatment with either fluticasone propionate aqueous nasal spray (FPANS) or placebo, 20 patients, allergic to HDM, registered symptoms (nasal obstruction, rhinorrhoea, sneezing, pruritus and eye symptoms) using three different scoring methods [Lebel, categorical and visual analogue scale (VAS)] and peak nasal inspiratory flow (PNIF) after HDM provocations. Provocations were performed with 1000 biological units/ml and 24 h later with 10,000 biological units/ml of HDM. Before and after the provocations, nasal mucosa biopsies were taken for immunohistochemical staining to determine the number of eosinophils. RESULTS: House dust mite provocations resulted in an increase in symptoms and a decrease in PNIF. Even at high-dose provocation, the FPANS group registered significantly lower symptoms than the placebo group for nasal blockage, sneezing, eye symptoms, and PNIF in both early and late phases. FPANS also suppressed rhinorrhoea during the late phase and the influx of eosinophils in the lamina propria. CONCLUSION: Despite the high background of symptoms, allergic responses can be induced in this perennial rhinitis model. The VAS score seems most suited to detect these changes and the suppression of symptoms by 7 days of FPANS treatment. Epithelial eosinophilia at baseline was correlated positively with the severity of the reaction after the first provocation. PMID- 15876305 TI - Polymorphism of tandem repeat in promoter of 5-lipoxygenase in ASA-intolerant asthma: a positive association with airway hyperresponsiveness. AB - BACKGROUND: 5-Lipooxygenase (ALOX5) and 5-lipoxygenase-activating protein (ALOX5AP) are known as key enzymes in cysteinyl-leukotriene (cys-LT) production, critical mediators in aspirin acetylsalicyclic acid (ASA)-intolerant asthma (AIA). To date, studies of the promoter region of ALOX5 gene has revealed the potential influence of a variable number of tandem repeats of a Sp1- and Egr1 binding motif, on the transcription rate. METHODS: To understand the pathological process that arises from cys-LT overproduction in AIA, we genotyped ALOX5 Sp1 and ALOX5AP poly(A) repeat promoter polymorphism by fluorescent-based capillary electrophoresis in the Korean population. RESULTS: No significant differences in allele and genotype frequencies of the ALOX5 and ALOX5AP promoter polymorphisms were observed between the three groups. However, there was a strong association of the ALOX5 Sp1 repeat polymorphism with airway hyperresponsiveness (AHR; PC20 methacholine); AIA patients carrying a mutant allele (n > 5 or n < 5 repeats) showed increased AHR compared to AIA patients with wild-type genotype (P=0.003). CONCLUSION: Although the alleles of the ALOX5 and ALOX5AP promoter cannot be considered as a prominent risk factor in the development of AIA, the genetic variant of tandem repeat (GGGCGG; Sp1-binding motif) in ALOX5 promoter is associated with the severity of airway hyperresponsiveness in AIA patients. PMID- 15876306 TI - Longitudinal study on the relationship between cat allergen and endotoxin exposure, sensitization, cat-specific IgG and development of asthma in childhood- report of the German Multicentre Allergy Study (MAS 90). AB - BACKGROUND: Controversial data have emerged regarding the question whether cat exposure in childhood favours or decreases the risk of sensitization and allergic airway disease. In a prospective birth-cohort study, we assessed the association between longitudinal cat allergen exposure, sensitization (immunoglobulin E, IgE), IgG antibody (ab) levels to cat and the development of asthma in children up to the age of 10 years. METHODS: Of 1314 newborn infants enrolled in five German cities in 1990, follow-up data at age 10 years were available for 750 children. Assessments included yearly measurements of specific serum IgE to cat and at age 6 and 18 months, 3, 4 and 10 years measurement of cat allergen Fel d 1 in house dust samples. Additionally, Fel d 1-specific IgG ab were determined in 378 serum samples of 207 children. Endotoxin exposure in mattress dust was measured in a subgroup of 153 children at age 10 years. From age 4 years on, International Study of Asthma and Allergy in Childhood (ISAAC) questionnaires were completed yearly in order to assess the prevalence of wheeze and asthma. RESULTS: Serum IgG-levels to cat showed a large variation, however, intraindividually values showed rather constant concentration over a longer time period. The IgG levels at school-age correlated with cat allergen exposure during the first 2 years of life. Specific IgE to cat was clearly associated with wheeze ever, current wheeze and bronchial hyperresponsiveness (BHR), this was also observed for children with specific IgE ab to cat (>0.35 kU/l) plus IgG levels above 125 U/ml. A large percentage of very highly exposed children showed high IgG but no IgE responses to cat, however, not all highly exposed children were found to be protected from sensitization. Children with IgG but without IgE ab to cat showed the lowest prevalence of wheeze ever and current wheeze despite high cat allergen exposure, however, this trend did not achieve significance. While homes of cat owners showed higher Fel d 1 concentrations than homes without cats, homes of cat owners were not found to have higher endotoxin levels in carpet dust samples than homes without cats. CONCLUSIONS: We could confirm that high cat allergen exposure in a cohort with lower community prevalence of cats is associated with higher serum IgG and IgE levels to cat in schoolchildren. Sensitization to cat allergen (IgE) is a risk factor for childhood asthma. While exposure to cat allergen during infancy is associated with sensitization (IgE), only in the very highly exposed children the likelihood of sensitization (IgE) is decreased and high IgG levels to cat without IgE were associated with low risk of wheeze. However, cat-specific IgG ab levels did not protect children with IgE mediated sensitization from wheeze. PMID- 15876307 TI - Effects of allergen inhalation and oral glucocorticoid on serum soluble CTLA-4 in allergic asthmatics. AB - BACKGROUND: The serum soluble cytotoxic T lymphocyte associated antigen-4 (sCTLA 4) concentration is significantly elevated in patients with asthma, and sCTLA-4 concentration correlate with the severity of asthma. The aim of the present study was to investigate effects of allergen inhalation and oral glucocorticoid on concentration of serum sCTLA-4 in patients with allergic asthma. METHODS: Allergen inhalation challenge was conducted in allergic asthmatics with isolated early asthma response and those with dual asthma response. In a randomized, double-blind, placebo-controlled, parallel group fashion, prednisolone or placebo was give orally once a day for 2 weeks. Venous blood samples were collected before and after allergen inhalation or prednisolone administration for obtaining sera. The serum sCTLA-4 concentrations were determined using enzyme-linked immunosorbent assay. RESULTS: The serum sCTLA-4 concentrations in the dual responder group increased from 29.0 (14.5-43.7) microg/l [median (25-75 percentiles)] before allergen inhalation to 44.0 (24.3-61.3) microg/l 24 h after allergen inhalation. In the isolated early responders, there were no significant increase in serum sCTLA-4 concentrations after allergen inhalation compared with baseline levels. There was a significant decrease in serum sCTLA-4 concentrations after 2 weeks of glucocorticoid therapy [22.0 (15.5-31.0) microg/l] compared with baseline values [37.0 (19.5-53.0) microg/l], whereas there was no significant difference in the placebo group. CONCLUSION: This study has demonstrated that serum sCTLA-4 concentrations increased after allergen inhalation in sensitized asthmatic subjects, and that serum sCTLA-4 concentrations were downregulated by prednisolone therapy. PMID- 15876308 TI - Polarization of Th1/Th2 in human CD4 T cells separated by CD62L: analysis by transcription factors. AB - BACKGROUND: T-cell surface antigens that differentiate clearly between Th1 and Th2 have not been identified. Discrimination of Th1/Th2 subpopulations by CD62L expression has been reported. We investigated the expression of transcription factors that regulate Th1/Th2 cytokine synthesis in human CD4+ T-cell subpopulations separated by CD45RO and CD62L, and compared the ratio of CD62L+ to CD62L- cells between healthy individuals and patients with allergic diseases. METHODS: Human peripheral blood samples were obtained from healthy volunteers and patients. CD4+ T cells were isolated by negative selection. Three CD4+ T-cell subpopulations separated by CD45RO and CD62L were isolated using three-color fluorescence. Sorted cells were stimulated with anti-CD3 monoclonal antibody, and the cytokine levels were measured using a Cytometric Bead Array Kit. Transcription factor expression was examined by reverse transcriptase polymerase chain reaction (RT-PCR) and real-time RT-PCR. RESULTS: Interleukin (IL)-4 and IL 5 production levels by CD45RO+CD62L+CD4+ T cells were higher than those of CD45RO+CD62L-CD4+ T cells (P < 0.05), whereas interferon-gamma and tumor necrosis factor-alpha production were lower levels (P < 0.05). T-cell immunoglobulin mucin 3 and T-bet expression were detected in CD45RO-CD62L+ and CD45RO+CD62L- cells following stimulation, but not in CD45RO+CD62L+ cells. However, the ratio of CD62L+ to CD62L- cells was the same in both healthy individuals and patients (P = 0.54). There was no difference in Th1/Th2 cytokine synthesis by CD4+ T cells. CONCLUSION: Analyses of cytokine syntheses and transcription factor expression demonstrated that CD62-negative and -positive subpopulations of human CD45RO+CD4+ T cells represent characteristics of Th1 and Th2, respectively. PMID- 15876309 TI - Costs associated with persistent allergic rhinitis are reduced by levocetirizine. AB - BACKGROUND: Allergic rhinitis was recently classified by the ARIA guidelines as persistent or intermittent. Levocetirizine was shown to improve symptoms and health-related quality of life of patients with persistent allergic rhinitis in the XPERT study, a 6-month randomized double blind placebo-controlled trial. OBJECTIVE: To assess the total costs of persistent allergic rhinitis, and the effect of long-term treatment with levocetirizine on these costs from several perspectives (societal, social security system, and employers). METHODS: Direct medical cost parameters (medications, physician visits and hospitalizations) and time lost parameters (workdays and Usual Daily Activities (UDA) lost) related to persistent allergic rhinitis and its comorbidities (asthma, sinusitis, otitis and upper respiratory infection) were measured. A cost analysis was performed using 2002 French costing data. RESULTS: From a societal perspective, the total cost of persistent allergic rhinitis without long-term treatment was estimated at 355.06/patient/month. From a social security perspective, levocetirizine treatment yielded an additional cost of 2.78/patient/month, compared to no treatment. However, levocetirizine reduced the total cost of persistent allergic rhinitis and its comorbidities by 152.93/patient/month from a societal perspective and by 64.70/patient/month from an employer perspective. Most gains resulted from a decrease in the lost workdays and UDA in the levocetirizine group. CONCLUSION: The cost of persistent allergic rhinitis is substantial. Treatment with levocetirizine reduces the cost of persistent allergic rhinitis and its comorbidities to the society by 152.93/patient/month while improving symptoms and health-related quality of life. PMID- 15876310 TI - Repeatability of peak nasal inspiratory flow measurements and utility for assessing the severity of rhinitis. AB - BACKGROUND: The measurement of peak nasal inspiratory flow (PNIF) provides a simple, cheap, fast and readily available tool for determining the extent of nasal airway patency. However, there are questions regarding its repeatability when used to assess the degree of nasal obstruction in large populations. Therefore, this study aimed to evaluate the repeatability of PNIF measurements and to assess their association with the signs and symptoms of rhinitis. METHODS: The PNIF, rhinitis symptoms, judged by Meltzer questionnaire and rhinitis signs, as determined by anterior rhinoscopy, were assessed in 283 adults representative of the general population. One training and two test PNIF measurements were recorded during the same session. RESULTS: The PNIF was highly reproducible (ICC = 0.92; 95% limits of agreement: +/-36 l/min). The PNIF was strongly correlated with rhinitis signs, measured by anterior rhinoscopy (rs= -0.38, P < 0.0001) but was not correlated with rhinitis symptoms, measured by questionnaire (rs= -0.11, P = 0.057). Differences in PNIF for subjects categorized as asymptomatic, mild or moderate/severe on the basis of rhinitis signs, were highly significant (P < 0.0001), but less significant on the basis of rhinitis symptoms (P = 0.04). A PNIF cut-off of 115 l/min had moderately high specificity (72%) and sensitivity (65%) and a high negative predictive value (90%) for moderate/severe signs of rhinitis. CONCLUSION: In a large general population-based sample of young adults, PNIF was highly reproducible and closely related to the signs of rhinitis, as determined by clinical examination. The PNIF provides information that is qualitatively different to that provided by symptom scores and may be useful to measure the extent of nasal obstruction. PMID- 15876311 TI - Efficacy and safety of preseasonal-specific immunotherapy with an aluminium adsorbed six-grass pollen allergoid. AB - BACKGROUND: The clinical efficacy and safety of a six-grass pollen allergoid has been studied. The advent of more exacting clinical guidelines and a better appreciation of the possible mechanisms of treatment prompted this reappraisal. METHODS: A 2-year double-blind multicentre placebo-controlled phase 3 clinical trial was undertaken in 154 patients suffering symptoms of rhinoconjunctivitis with or without asthma (GINA I or II). Therapy comprised two consecutive preseasonal short-courses of subcutaneous injections using a grass pollen allergoid adsorbed to aluminium hydroxide. RESULTS: A combined symptom and medication score (SMS) was used as the primary end-point for clinical efficacy. SMS from the first year showed a significant difference of 26.6% between the two study groups (P=0.026) and this was improved after the second year when there was a 48.4% difference in SMS between active and placebo treatment in favour of the allergoid (P = 0.018). Highly significant increases in grass pollen allergen specific IgG1 and IgG4 antibody concentrations were measured in association with active treatment. Allergen tolerance was increased as judged by a conjunctival provocation test and significant improvements in quality of life were documented using a standardized questionnaire. The allergoid was well tolerated. CONCLUSIONS: The grass pollen allergoid was shown to be safe and clinically efficacious in the management of hay fever with or without asthma (GINA I or II). PMID- 15876312 TI - Identification of an obeche (Triplochiton scleroxylon) wood allergen as a class I chitinase. AB - BACKGROUND: Wood dust is known to cause allergic occupational asthma and obeche (Triplochiton scleroxylon) is a prominent exponent in this field. However, the knowledge about wood allergens is still limited. The aim of this study was to identify and characterize obeche wood allergens. METHODS: Obeche extracts were prepared from freshly ground in comparison to 7 years stored wood dust and investigated by Sodium dodecyl sulfate-polyacrylamid gel electrophoresis, enzyme linked allergosorbent test and immunoglobulin (Ig)E-immunoblot. Allergens were detected by specific IgE of seven obeche allergic patients' sera and protein analysis was performed by mass spectrometry. Cross-reactivity was demonstrated by ImmunoCAP-inhibition with sera of seven obeche and four latex-allergic patients. RESULTS: Obeche extracts showed different protein pattern and IgE-binding capacities depend on the age of the wood dust. A 38 kDa protein was identified as major obeche wood allergen, detected by six of seven (85%) obeche allergic patients' sera and was entitled as Trip s 1. Trip s 1 is homologous to plant class I chitinases and exhibited enzyme activity demonstrated by chitinolysis. Co recognition or cross-reactivity of Trip s 1 according to structural similarity was seen in sera of latex allergic patients. IgE inhibition studies with obeche as solid phase and Trip s 1 and latex hevein as inhibitor demonstrated that Trip s 1 was a more effective inhibitor in obeche as well as in latex allergic patients' sera. CONCLUSIONS: Trip s 1 is a new obeche wood allergen of the plant class I chitinase family. This finding may explain the dominant role of obeche in sensitization against wood dust. PMID- 15876313 TI - Identification of IgE-binding epitopes on gliadins for patients with food allergy to wheat. AB - BACKGROUND: Food allergy to wheat induces different symptoms as atopic eczema/dermatitis syndrome (AEDS), urticaria and more severe reactions as wheat dependent exercise-induced anaphylaxis (WDEIA). Different gliadin classes are involved in this allergy but IgE-binding epitopes are known only on omega5 gliadins and for WDEIA cases. OBJECTIVES: The aim of the study was to identify IgE-binding epitopes on several gliadin classes and for several patients with different symptoms and ages. METHODS: Eleven sera were analysed by pepscan with overlapping synthetic peptides. RESULTS: Sera from five patients with anaphylaxis, urticaria or WDEIA, displayed strong IgE-binding to sequential epitopes of the repetitive domains of alphabeta, gamma, omega2 or omega5-gliadins with two immunodominant epitopes on omega5-gliadin and a consensus motif of the type QQX1PX2QQ (X1 being L, F, S or I and X2 Q, E or G). One patient allergic to deamidated wheat proteins also had IgE to a repetitive peptide of gamma and omega2-gliadins of the type QPQQPFP. Sera from four patients with AEDS detected no linear epitopes on gliadins, despite the fact that they contained specific IgE to alpha, beta, gamma or omega-gliadins. One child with AEDS recognized cysteine containing sequences in the nonrepetitive domains of alphabeta and gamma gliadins. CONCLUSION: B epitopes in wheat allergy were different from B epitopes of coeliac disease. Differences exist in IgE-binding epitopes between patients with food allergy to wheat. IgE from those suffering from WDEIA, anaphylaxis and urticaria detected sequential epitopes in the repetitive domain of gliadins whereas IgE from AEDS patients probably recognized conformational epitopes. PMID- 15876314 TI - Epsilon germ-line and IL-4 transcripts are expressed in human intestinal mucosa and enhanced in patients with food allergy. AB - BACKGROUND: The mechanisms of gastrointestinal (GI) food allergy (FA) are poorly understood. Immunoglobulin E (IgE) is increased in stools from patients with FA, as well as the number of cells carrying IgE in intestinal mucosa, but the origin of IgE production remains unknown. To investigate a local production of IgE in intestine, we analysed the levels of transcripts for epsilon germ-line (epsilonGT), and potential regulators of IgE production, IL-4, IL-13, IFN-gamma, IL-4Ralpha, STAT6 and FcepsilonRIalpha in intestinal mucosa of adult patients with FA. METHODS: Endoscopic biopsies were obtained from the caecum of 25 patients with FA and 14 control patients. The levels of epsilonGT, IL-4, IL-13, IFN-gamma, IL-4Ralpha, STAT6 and FcepsilonRIalpha mRNA were analysed by real-time RT-PCR and compared with unpaired nonparametric Mann-Whitney test. RESULTS: The mean epsilonGT transcript level in caecum was increased in FA patients compared with control patients (P < 0.05). IL-4 mRNA expression was also increased in FA patients (P < 0.05), whereas mRNA expression for IL-13, IFN-gamma, IL-4Ralpha, STAT6 and FcepsilonRIalpha mRNA expression was not altered. However, the ratio of IL-4 mRNA/IFN-gamma mRNA was significantly increased in FA patients (P < 0.05). No correlation was observed between epsilonGT transcripts expression in intestinal mucosa and total IgE levels in serum. CONCLUSIONS: This study shows that (i) epsilonGT transcripts are expressed in human intestinal mucosa; (ii) epsilonGT and IL-4 transcripts are increased in caecal mucosa from patients with FA. These results suggest local production of IgE in intestine that might be of importance for inflammatory reactions in the GI tract. PMID- 15876315 TI - Anaphylaxis during anesthesia: results of a 12-year survey at a French pediatric center. AB - BACKGROUND: Following adverse reactions to anesthesia, tests are carried out to determine the mechanism of the reaction and to identify the agent responsible. No specific data are available in France concerning such skin tests in children. METHODS: Between 1989 and 2001, we assessed hypersensitivity reactions to general anesthesia in 68 children. Thirty underwent more than one operation, for congenital malformations. Immunoglobulin (Ig)E-mediated anaphylaxis was diagnosed on skin tests combined with the clinical history. RESULTS: Grade I, II and III reactions were observed in 20, 27 and 21 children, respectively. IgE-mediated anaphylaxis was diagnosed in 51 children: 31 (60.8%) for neuromuscular blocking agents (NMBA), 14 (27%) for latex, seven (14%) for colloids, five (9%) for opioids and six (12%) for hypnotics. Vecuronium was the NMBA causing the largest number of reactions. Cross reactivity to NMBA available in France was observed in 23 of 30 children (76%), particularly for vecuronium and atracurium or pancuronium. The estimated frequency of IgE mediated anaphylactic reactions was one in 2100 operations. Based on our results, 25 children subsequently received a different anesthetic with no adverse reaction. CONCLUSIONS: As in adults, NMBA, then latex were responsible for most anaphylactic reactions during anesthesia. Our results confirm that skin tests with anesthetic agents are feasible and safe in children and improve the safety of subsequent anesthetic procedures. PMID- 15876316 TI - CA 125: marker of serosal inflammation in hereditary angioedema? PMID- 15876317 TI - The association of selective IgA deficiency and IgE hypogammaglobulinemia. PMID- 15876318 TI - Microwave radiation from cellular phones increases allergen-specific IgE production. PMID- 15876319 TI - Occupational allergy to raw beef due to cross-reactivity with dog epithelium. PMID- 15876320 TI - Analysis of the crown fractures and crown-root fractures due to dental trauma assisted by the Integrated Clinic from 1992 to 2002. AB - The purpose of this study was to analyze crown fractures and crown-root fractures due to dentoalveolar trauma, treated in the Integrated Clinic comprehensive dental care at Aracatuba School of Dentistry (UNESP), from January 1992 to July 2002. The data were obtained from files of trauma cases. On the analysis period, 293 patients had crown fractures or crown-root fractures, in 605 teeth. Sixty nine percent were males and 31% were females. Adolescents between 11 and 18 years old were the most prevalent group (41.6%) and the maxillary arch was the most commonly traumatized (83%). The most commonly affected tooth was the maxillary central incisor (58.3%). The most frequent causes were falls from bicycles (30.8%). It was concluded that the reality of the local service is similar to the published data. PMID- 15876321 TI - Oro-facial injuries in Central American and Caribbean sports games: a 20-year experience. AB - Dental services in sports competitions in the Games sponsored by the International Olympic Committee are mandatory. In every Central American, Pan American and Olympic Summer Games, as well as Winter Games, the Organizing Committee has to take all the necessary measures to assure dental services to all competitors. In all Olympic villages, as part of the medical services, a dental clinic is set up to treat any dental emergency that may arise during the Games. Almost every participating country in the Games has its own medical team and some may include a dentist. The major responsibilities of the team dentist as a member of the national sports delegation include: (i) education of the sports delegation about different oral and dental diseases and the illustration of possible problems that athletes or other personnel may encounter during the Games, (ii) adequate training and management of orofacial trauma during the competition, (iii) knowledge about the rules and regulations of the specific sport that the dentist is working, (iv) understanding of the anti-doping control regulations and procedures, (v) necessary skills to fabricate a custom-made and properly fitted mouth guard to all participants in contact or collision sports of the delegation. This study illustrates the dental services and occurrence of orofacial injury at the Central American and Caribbean Sports Games of the Puerto Rican Delegation for the past 20 years. A total of 2107 participants made up the six different delegations at these Games. Of these 279 or 13.2% were seen for different dental conditions. The incidence of acute or emergency orofacial conditions was 18 cases or 6% of the total participants. The most frequent injury was lip contusion with four cases and the sport that experienced more injuries was basketball with three cases. PMID- 15876322 TI - Evaluation of intruded primary incisors. AB - Tooth intrusion consists of the displacement of the tooth into its alveolus and is the most common trauma during early infancy. This work aims to evaluate the aspects related to tooth intrusion in primary teeth by monitoring 16 patients (22 teeth) for a period between 3 and 36 months. Of the patients who suffered from tooth intrusion, 56.25% were male and 91% of the intruding teeth were upper central incisors. In all cases the treatment indicated was to await spontaneous re-eruption: total re-eruption occurred in 42.5% of cases, partial re-eruption in 47% of cases and in 10.5% there was no re-eruption. Twenty-three percentage of the teeth suffered necrosis, 33% suffered internal or external root resorption and none suffered root canal obliteration. Fifty-seven percentage indicated healthy pulps independent of degree of re-eruption. PMID- 15876323 TI - Can mouthguards prevent mandibular bone fractures and concussions? A laboratory study with an artificial skull model. AB - Some sports' accidents are responsible for inflicting traumatic brain injuries and mandibular bone fractures when impacts occur to the chin. It is often thought that mouth guards can prevent many of these injuries. However, such assertions may be insufficient without adequate research. It is therefore necessary to establish a systematic method of investigation to solve this problem. In the present laboratory study, tests were performed using pendulum impact equipment and an artificial skull model connected to strain gages and accelerometers to simulate and measure the surface distortions related to bone deformation or fractures and the acceleration of the head related to concussions. As impacts, direct blows to the mandibular undersurface were applied. As a result, wearing a mouth guard decreased (P < 0.01) the distortion to the mandibular bone and the acceleration of the head significantly compared with not wearing a mouth guard (54.7%: to the mandible -- measured at a total of three different points, 18.5%: to the head measured at a total of three different points). Within the limits of this study, the following conclusions were drawn: The present measuring system in this study was able to evaluate the distortion to the mandibular and the acceleration of the head from the direct blow to the mandibular undersurface. Mouth guards can reduce distortion to the mandibular and the acceleration of the head from the same blow. So mouth guards might have the possibility to prevent mandibular bone fractures and concussions. However, further well-designed and exhaustive studies are vital to show that mouth guards reduce the incidence of concussions and mandibular bone fractures. PMID- 15876324 TI - Study of calcium hydroxide apexification in 26 young permanent incisors. AB - This study was carried out in order to observe the effectiveness of apexification in young permanent incisors. The sample comprised 26 young permanent incisors with necrotic pulp and open apices. The time taken to obtain apical closure, its form and size were analysed in order to find out if closure was influenced by existing pathology or size of apex. The treated teeth were compared with their corresponding contralateral teeth. The test of McNemar and anova was used and a result of P = 0.05 was considered significant. Teeth with pretreatment apical shapes that were convergent or parallel all resulted in physiological apical shapes after treatment. Eight teeth had divergent apical shapes before treatment. Of these, one had a physiological shape, five ended with rounded apices and two teeth had straight apices post-treatment. Apical closure was obtained in 100% of the cases studied, of these 88.4% needed three to four sessions of calcium hydroxide treatment (an average of 3.23 sessions) in order to obtain apical closure, the average time employed was 12.19 months. Clinical symptoms resolved in all teeth that presented with symptoms. Preoperative symptoms did not affect outcome. Pathology of the tooth before treatment does not influence the time needed to obtain apical closure. PMID- 15876325 TI - Dentinal pH changes following electrophoretically activated calcium hydroxide ions in the root canal space of bovine teeth. AB - The aim of the study was to evaluate pH changes of external root dentin surface at different depths following electrophoretic activation of calcium hydroxide (CH) in the root canals. For the study, 60 cavities were drilled on three external root surfaces of 20 bovine teeth specimens to obtain remaining dentin thickness (RDT) of 0.3, 0.7 and 1.0 mm. CH paste was inserted in the lumens. In the experimental group (10 specimens) CH paste was electrophoretically activated. Microelectrode was used to measure pH changes in the cavities immediately after placement of CH, following electrophoretical activation, and after 30 days storage. A significant (P < 0.05) increase in pH following electrophoresis was found in all specimens in the experimental group. Cavities with 0.4 mm RDT showed a maximal pH increase to the value of 11.3 +/- 0.4. In the control group no change in pH was observed after 3 min. A similar increase in pH was observed in both groups after 30 days. The pH changes were significantly depended on the RDT (P < 0.05) in all groups. Electrophoretically activated CH could significantly increase dentinal pH of external root surface within minutes up to 30 days. PMID- 15876326 TI - A comparative study of white and grey mineral trioxide aggregate as pulp capping agents in dog's teeth. AB - Mineral trioxide aggregate (MTA) is widely used for different perforation repairs, root-end filling, pulp capping and many other endodontic procedures. Because of potential discoloration effect of gray MTA, white MTA has been introduced into endodontic treatment for the same purposes. This study examined the dental pulp responses in dogs to both types of MTA used as a pulp-capping agent. The pulps of 24 teeth of four male dogs were exposed with a No.1 round bur and capped with either gray or white MTA. Histologic analysis was performed one and 2 weeks after treatment. Calcified bridge could be seen 1 week after treatment with both types of MTA, with no significant differences between the two treatments. PMID- 15876327 TI - Structural resistance in immature teeth using root reinforcements in vitro. AB - The purpose of this study was to evaluate in vitro the efficacy of root reinforcements by light-cured composite resin or zirconium fiber post in simulated immature non-vital teeth. Fifty-six bovine incisors teeth were used for this study. The crown of each tooth was removed in the medium third to obtain a standard length of 30 mm. The specimens were divided into four groups (n = 14): G1) the root canals were instrumented and enlarged to simulate immature non-vital teeth and were reinforced with a light-cured composite resin using a translucent curing post (Luminex system); (G2) the specimens were instrumented, enlarged and they received root reinforcement with zirconium fiber post; G3 (positive control): they received similar treatment to the G1 and G2 groups, but did not receive root reinforcement; G4 (negative control): the roots were not weakened and did not receive reinforcement. Every tooth was submitted to compressive force using an Instron testing machine with an angle of 45 degrees at a speed of 1 mm min(-1) until the fracture. The results showed a markedly increased resistance to fracture in the G1 and G2 (122.38 and 122.08 kg f, respectively). Among the results of G1 and G2 there was not any significant difference (P > 0.05) but they were significantly different from the control groups (P < 0.05). The conclusion is that the use of root reinforcements with zirconium fiber post or composite resin can increase significantly the structural resistance of the weakened teeth, decreasing the risk of the fracture. PMID- 15876328 TI - Orthodontic movement of a horizontally fractured tooth: a case report. AB - Intra-alveolar root fractures are relatively uncommon, and sometimes may be healed without treatment. Diagnosis of intra-alveolar root fracture is frequently made by exclusion of clinical signs of other traumatic injury, and by radiographic examination with films taken at different tube angulations. This case report presents a horizontal root fractured tooth that was healed satisfactory without treatment and moved orthodontically after prolonged time from trauma. PMID- 15876329 TI - Forced eruption after crown/root fracture with a simple and aesthetic method using the fractured crown. AB - Forced eruption offers a method of treatment of teeth fractured close to the alveolar crest. We introduce a modification of this common technique. The fractured fragment of a patient's crown is bonded to the adjacent teeth and used as anchorage, permitting the root extrusion while offering acceptable aesthetics. The bonded crown does not impede the eruption of the root so that the frequent occlusal adjustment of other methods is not required. The final result is acceptable and a low cost alternative to common techniques. PMID- 15876330 TI - The fate of a mid-root fracture: a case report. AB - The present report describes a case of a mid-root fracture in a maxillary central incisor of a 19-year-old patient. The fractured tooth was splinted with composite that was removed only 3 years later, as the patient did not appear for follow-up examinations. At this time, the radiographs revealed a normal periodontal ligament, rounding of the borders of the fragments and pulp obliteration of both fragments. Eight years later, the tooth was clinically normal and blurred calcification of the root canal was disclosed radiographically. After 13.5 years the patient complained of tooth mobility and radiographic examination revealed an advanced cervical root resorption. As no conservative approach was possible at this stage, the patient was referred to a prosthodontist for esthetic rehabilitation. PMID- 15876331 TI - Orthodontic extrusion of subgingivally fractured incisor before restoration. A case report: 3-years follow-up. AB - Orthodontic forced eruption may be a suitable approach without risking the esthetic appearance in tooth fracture below the gingival attachment or alveolar bone crest. Extrusion of such teeth allows elevating the fracture line above the epithelial attachment and so the proper finishing margins can be prepared. Restoration after orthodontic eruption may present a more conservative treatment choice in young patients compared with the prosthetic restoration after extraction. This case describes a multidisciplinary approach using the orthodontic forced eruption facilitating the composite restoration of a fractured upper permanent central incisor. PMID- 15876332 TI - An alternative method for fabricating a custom-made metal post with a ceramic core. AB - The restoration of anterior non-vital teeth with metal posts and cores and all ceramic restorations may lead to compromised esthetics because of the semi translucence of ceramics and the metallic color of the underlying post and cores. This article presents a technique that combines the optical properties of ceramic cores with the mechanical properties of custom-made cast metal posts. The technique involves heat pressing of the core from leucite-enriched glass-ceramic to the underlying custom-made metal post and may provide additional esthetic benefit for anterior teeth especially with little remaining coronal dentin and with small root volume. PMID- 15876333 TI - Multiple cerebral aneurysms as delayed complication of left cardiac myxoma: a case report and review. AB - Left cardiac myxoma and also consecutive embolization into the brain is well documented, whereas the association of myxomas with multiple fusiform cerebral aneurysms is rare. We analyze 33 previously reported patients and present a case of a 43-year-old woman with multiple cerebral infarctions 2 years after resection of a recurrent myxoma in the left atrium. Cerebral angiography displayed multiple fusiform aneurysms of several cerebral arteries, including a giant aneurysm of the basilar artery. Serum level of interleukin-6 (IL-6) was highly elevated. The clinical, radiological and pathological features of these aneurysms are summarized. The pathogenesis, including the role of IL-6 in the formation of myxomatous aneurysms, is discussed. PMID- 15876334 TI - G309D and W437OPA PINK1 mutations in Caucasian Parkinson's disease patients. AB - OBJECTIVE: To determine whether the G309D and W437OPA mutations in PINK1 gene are present in American Caucasian population of patients with Parkinson's disease (PD). METHODS: We searched for the G309D and W437OPA mutation by sequencing the regions of interest in the PINK1 gene in 237 unrelated Caucasian patients. RESULTS: None of the 237 samples showed the G309D or W437OPA mutations. CONCLUSIONS: The G309D and W437OPA mutations in PINK1 gene probably do not represent common causes of familial or sporadic PD in a Caucasian population. PMID- 15876335 TI - Can Alzheimer's type pathology influence the clinical phenotype of Parkinson's disease? AB - OBJECTIVES: Patients with clinical and pathological diagnosis of Parkinson's disease (PD) may, at death, also be found to have the pathological changes of Alzheimer's disease (AD). With this study we aim to determine the influence of AD pathology on the clinical phenotype of PD. METHODS: We studied 64 patients who donated their brains to the University of Miami Brain Endowment Bank(TM) and fulfilled the clinical and pathological criteria for PD. For the evaluation of AD pathology we used the CERAD criteria. Dementia was diagnosed, in life, also using standard criteria. Case histories were abstracted and reviewed by one investigator (SP) who then made comparisons between patients. RESULTS: Patients with AD pathology (PD-AD) were older both at the time of diagnosis and death. The presence of AD pathology did not seem to influence disease duration in our cohort of PD patients. As expected there was a clear relation between AD pathology and dementia but not all PD-AD patients were demented. Psychosis and depression were also found to be more prevalent in the PD-AD patients. In the comparison between demented and non-demented PD-AD patients dementia was more likely to appear in patients with PD and definite criteria for AD. CONCLUSION: Apart from dementia AD pathology seems to be associated with a number of other clinical characteristics of PD. PMID- 15876336 TI - IL-6: an early marker for outcome in acute ischemic stroke. AB - OBJECTIVES: Inflammation plays an important role in the pathophysiology of stroke. We correlated interleukin (IL)-6, IL-10, C-reactive protein (CRP) and T lymphocyte subtype levels in acute ischemic stroke patients with stroke volume and clinical outcome. MATERIALS AND METHODS: Blood samples were obtained from 11 patients at defined intervals during 1 year. Nine healthy age-matched subjects served as controls. IL-6, IL-10 and CRP were quantified by enzyme-linked immunosorbent assay and T lymphocytes by flow cytometry. Volume measurement was carried out by computed tomography or magnetic resonance imaging and clinical outcome was scored by the European stroke scale (ESS) and Barthel index (BI). RESULTS: IL-6 levels were increased in the acute phase of stroke compared with healthy controls (P = 0.002) and correlated with larger stroke volume (P = 0.012) and less favorable prognosis after 1 year, measured by ESS (P = 0.014) and BI (P = 0.006). IL-10, CRP and T-lymphocyte subtypes in the acute phase were not correlated with stroke volume or clinical outcome. CONCLUSION: IL-6 seems to be a robust early marker for outcome in acute ischemic stroke. PMID- 15876337 TI - Recovery of somatosensory deficits in acute stroke. AB - OBJECTIVES: To study the recovery of somatosensory deficits after acute stroke. MATERIAL AND METHODS: A detailed clinical examination of sensation, median nerve somatosensory evoked potentials (SEP), quantitative sensory tests (QST), and subjective evaluation were performed in five acute stroke patients at three control time points up to 12 months after the stroke. RESULTS: The deficit recovered at least partially in all patients, mostly within 3 months after stroke. The improvement in warm and vibration detection thresholds occurred between 3 and 12 months. The SEP improved both by 3 and 12 months. CONCLUSION: The recovery of subjective sensory disturbance occurred in line with the improvement of the clinical sensory tests and QST. The most sensitive measure for somatosensory dysfunction at the early phase was graphesthesia. In our patients, initially normal SEP with a sensory deficit resulted in excellent clinical recovery, whereas initially absent SEP did not necessarily predict poor outcome. PMID- 15876338 TI - One-legged bicycling as an assessment tool for patients with stroke. AB - OBJECTIVE: To assess whether one-legged bicycling correlates with muscle strength and thereby could work as an outcome measure for persons with stroke. METHODS: The study comprised 29 men (age 35-65) with a first occurrence of stroke 6-35 months earlier. Each leg was evaluated separately. A ramp protocol was used (10 W/min), with continuous recording of the ventilatory uptake (Vo(2)) and heart rate. An isokinetic dynamometer was used to assess strength and endurance. Enzyme assays were performed on muscle biopsy samples. RESULTS: The peak isometric strength and isokinetic strength of the paretic leg correlated with the max. W on the bicycle. The oxidative enzyme citrate synthase correlated with the workload for both legs on the bicycle and lactate dehydrogenase correlated with peak isometric strength in both legs. CONCLUSIONS: The one-legged bicycle exercise test can be used to assess endurance in persons with a previous stroke as it correlates with dynamometer testing and muscle biopsies. PMID- 15876339 TI - Lumbar infusion test in normal pressure hydrocephalus. AB - OBJECTIVE: To compare potential clinical value of plateau pressure (P(pl)), resistance to outflow (R(out)), pulse-pressure amplitude (P(plA)) and rate of pressure increase (v(P)), taken from the constant rate lumbar infusion test (LIT), as predictors for the outcome of shunt surgery. METHODS: Recordings from preoperative LIT in 55 patients were scrutinized for the values of P(pl), P(plA), v(P) and R(out). Gait, memory, spatial capacity and reaction ability were tested before and 6 months after shunt surgery. RESULTS: Forty-three (78%) of the patients improved. There were no statistically significant differences in P(pl), R(out), P(plA) or v(P) between improved and not improved patients. Five patients with P(pl) below 22 mmHg (the cut off level) improved after shunting, while 16 and eight patients with R(out) below the cut off levels of 18 and 14 mmHg/ml/min improved. P(plA) correlated with P(pl) and R(out) (r = 0.74 and 0.63, respectively). In the group of patients with high P(plA) (>/=20 mmHg) as many as 93% improved but a high P(plA) did not recruit more improved patients than P(pl) or R(out) alone. CONCLUSION: v(P) or P(plA) does not add useful information to P(pl) for selecting patients with suspected NPH for surgery. R(out) calculations from LIT does not provide advantage over using the steady-state plateau pressure for selecting patients for surgery and may increase the risk of missing patients who should benefit from surgery. PMID- 15876340 TI - Clinical and molecular events in patients with Machado-Joseph disease under lamotrigine therapy. AB - OBJECTIVE: Machado-Joseph disease (MJD)/spinocerebellar ataxia type 3 is an autosomal dominant spinocerebellar degeneration, for which there is no effective treatment. PATIENTS AND METHODS: This study involved the clinical response of lamotrigine (LTG) on six MJD patients with early truncal ataxia and the effect of LTG on the alteration of ataxin-3 expression in the transformed MJD lymphoblastoid cells. RESULT: LTG medication was found, on the basis of single leg standing test tandem gait index, to effectively improve gait balance, but did not prove to be effective in the withdrawal period. In Western blot analysis of ataxin-3 in MJD lymphoblastoid cells, extracellular application of LTG, while leaving the normal level of ataxin-3 intact, decreased the expression of mutant ataxin-3 in a dose-related manner. CONCLUSION: Our results indicated that LTG may have significant benefits in relief of gait disturbance in MJD patients with early ataxia, and may be related to the decreased expression of mutant ataxin-3. PMID- 15876341 TI - Autosomal dominant cerebellar ataxias in Spain: molecular and clinical correlations, prevalence estimation and survival analysis. AB - INTRODUCTION: The genetic and clinical profile of autosomal dominant cerebellar ataxias (ADCA) displays marked geographical and ethnical variability. MATERIALS AND METHODS: We have analysed the molecular and clinical correlations in an ethnically homogeneous sample of 30 Spanish ADCA kindreds. Minimal point prevalence for the region of Cantabria was estimated. RESULTS: Seventy per cent of the families harboured known mutations. Areflexia, slow saccades and hypopallesthesia predominated in SCA2; nystagmus, pyramidal signs or areflexia restricted to the legs in SCA 3; and retinal degeneration, pyramidal signs and slow saccades in SCA 7. Anticipation and intergenerational instability were greater in SCA 7. Length of expansions and age at onset were inversely correlated in all SCA subtypes. Larger expansions correlated with areflexia in SCA 2, with pyramidal signs in SCA 3 and with early visual impairment in SCA 7. Survival was similar among the different SCA subtypes. Prevalence of ADCA in Cantabria was 1.6 cases per 100,000 population. CONCLUSIONS: This report shows the epidemiological, clinical and genetic profile of ADCA in Spain, providing additional data regarding the broad clinical heterogeneity of these disorders and the variability of the genotype-phenotype correlations. PMID- 15876342 TI - Classification of fine-motoric disturbances in Wilson's disease using artificial neural networks. AB - Patients suffering from Wilson's disease are divided into several types according clinical symptoms only at time of manifestation. Thereby two main subgroups exist: neurologic and non-neurologic types. After long-term therapy the neurological symptoms occurring in hepatolenticular degeneration may be improved but frequently with remaining fine-motoric disturbances which should be used for evaluation of the actual patient state. These disturbances are difficult to assess in an exact and objective manner by clinical examination. Therefore we measured fine-motoric passive and active abilities based on a standardized test set using the VSCOPE-system. The parallel evaluation of all fine-motoric data using an artificial neural network leads to a reclassification of these patients based on actual fine-motoric abilities but not reflecting the clinical classification at time of manifestation. PMID- 15876343 TI - Balance disorders in headache patients: evaluation by computerized static stabilometry. AB - OBJECTIVES: The aim of the study was to investigate the occurrence of equilibrium disturbances in headache patients, during the interictal period, by computerized static stabilometry. MATERIALS AND METHODS: Sixty-seven patients were studied: 35 with migraine without aura (MwoA), 12 with tension-type headache (TTH) in the episodic form (ETTH) + MwoA, and 20 with only TTH [10 ETTH + 10 chronic (CTTH)]. The stabilometric parameters considered were: statokinesigram length (L) and surface (S) in open (EO) and closed (EC) eyes conditions with/without occlusal bite, EC with head retroflexion (ECR), and optokinetic stimulation (OKN). RESULTS: The alteration of at least one of the stabilometric parameters was observed in 45 patients (67.2%): 21 MwoA, 8 ETTH + MwoA, and 16 TTH (8 ETTH + 8 CTTH). CONCLUSION: In TTH patients (ETTH, CTTH, ETTH + MwoA), the stabilometric findings show a proprioceptive alteration induced by cervicofacial muscle contraction, which was peripheral in origin. In MwoA patients the alterations appear under OKN and support a control impairment in involuntary oculomotility of central origin. PMID- 15876345 TI - Incidence of stillbirth and perinatal mortality and their associated factors among women delivering at Harare Maternity Hospital, Zimbabwe: a cross-sectional retrospective analysis. AB - BACKGROUND: Death of an infant in utero or at birth has always been a devastating experience for the mother and of concern in clinical practice. Infant mortality remains a challenge in the care of pregnant women worldwide, but particularly for developing countries and the need to understand contributory factors is crucial for addressing appropriate perinatal health. METHODS: Using information available in obstetric records for all deliveries (17,072 births) at Harare Maternity Hospital, Zimbabwe, we conducted a cross-sectional retrospective analysis of a one-year data, (1997-1998) to assess demographic and obstetric risk factors for stillbirth and early neonatal death. We estimated risk of stillbirth and early neonatal death for each potential risk factor. RESULTS: The annual frequency of stillbirth was 56 per 1,000 total births. Women delivering stillbirths and early neonatal deaths were less likely to receive prenatal care (adjusted relative risk [RR] = 2.54; 95% confidence intervals [CI] 2.19-2.94 and RR = 2.52; 95% CI 1.63 3.91), which for combined stillbirths and early neonatal deaths increased with increasing gestational age (Hazard Ratio [HR] = 3.98, HR = 7.49 at 28 and 40 weeks of gestation, respectively). Rural residence was associated with risk of infant dying in utero, (RR = 1.33; 95% CI 1.12-1.59), and the risk of death increased with increasing gestational age (HR = 1.04, HR = 1.69, at 28 and 40 weeks of gestation, respectively). Older maternal age was associated with risk of death (HR = 1.50; 95% CI 1.21-1.84). Stillbirths were less likely to be delivered by Cesarean section (RR = 0.64; 95% CI 0.51-0.79), but more likely to be delivered as breech (RR = 4.65; 95% CI 3.88-5.57, as were early neonatal deaths (RR = 3.38; 95% CI 1.64-6.96). CONCLUSION: The frequency of stillbirth, especially macerated, is high, 27 per 1000 total births. Early prenatal care could help reduce perinatal death linking the woman to the health care system, increasing the probability that she would seek timely emergency care that would reduce the likelihood of death of her infant in utero. Improved quality of obstetric care during labor and delivery may help reduce the number of fresh stillbirths and early neonatal deaths. PMID- 15876346 TI - PKC translocation and ERK1/2 activation in compensated right ventricular hypertrophy secondary to chronic emphysema. AB - BACKGROUND: Right ventricular hypertrophy (RVH) is an important complication of chronic lung disease. However, the signal transduction pathways involved as well as the physiological changes to the right ventricle have not been investigated. Emphysema was produced in male, Syrian Golden hamsters by intra-tracheal instillation of 250 IU/kg elastase (Emp, n = 17). Saline treated animals served as controls (Con, n = 15). RESULTS: Nine months later, Emp hamsters had 75% greater lung volume, and evidence of RVH at the gross and myocyte level (RV:tibia length Emp 6.84 +/- 1.18 vs. Con 5.14 +/- 1.11 mg/mm; myocyte cross sectional area Emp 3737 vs. Con 2695 microm2), but not left ventricular hypertrophy. Serial echocardiographic analysis from baseline to nine months after induction of emphysema revealed increasing right ventricular internal dimension and decreased pulmonary artery acceleration time only in Emp hamsters. There was an increase in translocation of PKC betaI and PKC epsilon from cytosolic to membranous cell fractions in RV of Emp hamsters. Phosphorylation of PKC epsilon was unchanged. Translocation of PKC alpha and betaII were unchanged. Emp animals had a 22% increase in phospho-ERK 1/2, but no change in levels of total ERK 1/2 compared to Con. CONCLUSION: These data suggest that PKC betaI, epsilon and ERK 1/2 may play a role in mediating compensated RVH secondary to emphysema and may have clinical relevance in the pathogenesis of RVH. PMID- 15876347 TI - Racemic epinephrine compared to salbutamol in hospitalized young children with bronchiolitis; a randomized controlled clinical trial [ISRCTN46561076]. AB - BACKGROUND: Bronchiolitis is the most common cause of lower respiratory tract illness in infancy, and hospital admission rates appear to be increasing in Canada and the United States. Inhaled beta agonists offer only modest short-term improvement. Trials of racemic epinephrine have shown conflicting results. We sought to determine if administration of racemic epinephrine during hospital stay for bronchiolitis improved respiratory distress, was safe, and shortened length of stay. METHODS: The study was a randomized, double-blind controlled trial of aerosolized racemic epinephrine compared to salbutamol every one to 4 hours in previously well children aged 6 weeks to < or = 2 years of age hospitalized with bronchiolitis. The primary outcome was symptom improvement as measured by the Respiratory Distress Assessment Instrument (RDAI); secondary outcomes were length of stay in hospital, adverse events, and report of symptoms by structured parental telephone interview one week after discharge. RESULTS: 62 children with a mean age of 6.4 months were enrolled; 80% of children had Respiratory Syncytial Virus (RSV). Racemic epinephrine resulted in significant improvement in wheezing and the total RDAI score on day 2 and over the entire stay (p < 0.05). The mean LOS in the epinephrine arm was 2.6 days (95% CI 2, 3.2) v. 3.4 days in those in the salbutamol group (95% CI 2.6, 4.2) (p > 0.05). Adverse events were not significantly different in the two arms. At one week post-discharge, over half of parents reported that their child still had a respiratory symptom and 40% had less than normal feeding. CONCLUSION: Racemic epinephrine relieves respiratory distress in hospitalized infants with bronchiolitis and is safe but does not abbreviate hospital stay. Morbidity associated with bronchiolitis as identified by parents persists for at least one week after hospital discharge in most infants. PMID- 15876348 TI - Early life factors, childhood cognition and postal questionnaire response rate in middle age: the Aberdeen Children of the 1950s study. AB - BACKGROUND: Little is known about the relationship between early life factors and survey response in epidemiological studies of adults. METHODS: The Children of the 1950s cohort is composed of 12,150 children (boys 51.7%) born in Aberdeen 1950-56 and in primary schools in the city in 1962. Information on birth weight, gestational age, growth, behaviour and socio-economic position at birth and in childhood were obtained from contemporaneous records. Cognitive test scores at ages 7,9 and 11 years were also available from school records. The outcome was response to a postal questionnaire sent (2001-2003) to surviving cohort members in middle age. RESULTS: Of 11,282 potentially mailed subjects, 7,183 (63.7%) returned questionnaires. Response rates were highest among females, and those whose parents were married at birth, were in a non-manual social class at birth or in childhood, had fewer siblings, were taller and heavier in childhood for their age and had lower Rutter B behavioural scores. Childhood cognitive test scores at every age were strongly and positively related to the response rate to a postal questionnaire independently of other early life factors monotonically across the entire range of test scores. Those in the bottom fifth at age 11 had a response rate of 49% while those in the top fifth 75%. CONCLUSION: The strength and consistency of the association of childhood cognition with questionnaire response rate in middle age is surprisingly large. It suggests that childhood cognition across the entire normal range is a powerful influence on the complex set of later behaviours that comprise questionnaire response. The extent of possible response bias in epidemiological studies of the associations between childhood characteristics (particularly those related to cognition) and later health is probably larger than is generally realised, at least in situations where the survey instrument is a postal questionnaire. PMID- 15876349 TI - The first case of septicemia due to nontoxigenic Corynebacterium diphtheriae in Poland: case report. AB - BACKGROUND: Toxigenic strains of Corynebacterium diphtheriae are well known agent of diphtheria. Nontoxigenic strains can cause atypical course of the disease. Invasive diseases caused by C. diphtheriae occur very rare. CASE PRESENTATION: We have described the first case of septicemia and endocarditis due to nontoxigenic C. diphtheriae biotype gravis in Poland. The patient has not belonged to any group of risk such infection. CONCLUSION: The case presented in this article shows unusual case of infection connected with nontoxigenic C. diphtheriae that took place in the area where have been no cases of diphtheria and other C. diphtheriae infections for near ten years. It shows the importance of identifying Corynebacterium isolates at the species level especially when the strain has been isolated from normally sterile sites. PMID- 15876350 TI - Amplification and overexpression of the ID4 gene at 6p22.3 in bladder cancer. AB - BACKGROUND: Amplifications at 6p22.3 are prevalent in advanced stage bladder cancer (TCC). Previous studies have identified SOX4, CDKAL, and E2F3 as targets of this amplification and therefore potential oncogenes, but the more telomeric DEK gene too has been reported as overexpressed and amplified. We have therefore investigated whether the intermediate region harboring the oncogene candidate ID4 is also part of the amplicon. RESULTS: Expression of E2F3, DEK, and ID4 was investigated by real-time RT-PCR in 28 TCC compared to 6 normal bladder tissues and in 15 TCC cell lines compared to cultured normal urothelial cells. Expression of E2F3 as well as DEK increased on average in tumor vs. normal tissues (3-fold and 2.5-fold, resp.), but only the increase for E2F3 was statistically significant (p = 0.039). ID4 overexpression was observed in selected specimens. Each of the three genes was overexpressed in several cell lines, up to 150-fold (ID4), 30-fold (E2F3), and 9-fold (DEK), but these increases were not correlated to each other. Instead, moderate (DEK) to excellent (ID4) correlations were observed with copy number increases of microsatellites near each gene. Microsatellite copy number increases were highly heterogeneous across the investigated several Mb region revealing at least three subregions of amplification. CONCLUSION: Extending previous reports, our data indicate that the 6p22.3 amplicon in TCC is highly heterogeneous and targets several genes in a variable fashion. Among these, expression of E2F3 and DEK appear to be generally increased in TCC, with additional increases caused by amplifications. In contrast, over-expression of ID4, which is normally predominantly expressed in testes and brain, appears to depend more strictly on gene amplification. Accordingly, the effect of amplifications at 6p22.3 in bladder cancer is expected to be non-uniform, thereby contributing to the highly variable biological and clinical behavior of advanced stage tumors. ID4 is a potential oncogene in a small subset of bladder cancers. PMID- 15876351 TI - Diaspora, a large family of Ty3-gypsy retrotransposons in Glycine max, is an envelope-less member of an endogenous plant retrovirus lineage. AB - BACKGROUND: The chromosomes of higher plants are littered with retrotransposons that, in many cases, constitute as much as 80% of plant genomes. Long terminal repeat retrotransposons have been especially successful colonizers of the chromosomes of higher plants and examinations of their function, evolution, and dispersal are essential to understanding the evolution of eukaryotic genomes. In soybean, several families of retrotransposons have been identified, including at least two that, by virtue of the presence of an envelope-like gene, may constitute endogenous retroviruses. However, most elements are highly degenerate and are often sequestered in regions of the genome that sequencing projects initially shun. In addition, finding potentially functional copies from genomic DNA is rare. This study provides a mechanism to surmount these issues to generate a consensus sequence that can then be functionally and phylogenetically evaluated. RESULTS: Diaspora is a multicopy member of the Ty3-gypsy-like family of LTR retrotransposons and comprises at least 0.5% of the soybean genome. Although the Diaspora family is highly degenerate, and with the exception of this report, is not represented in the Genbank nr database, a full-length consensus sequence was generated from short overlapping sequences using a combination of experimental and in silico methods. Diaspora is 11,737 bp in length and contains a single 1892-codon ORF that encodes a gag-pol polyprotein. Phylogenetic analysis indicates that it is closely related to Athila and Calypso retroelements from Arabidopsis and soybean, respectively. These in turn form the framework of an endogenous retrovirus lineage whose members possess an envelope-like gene. Diaspora appears to lack any trace of this coding region. CONCLUSION: A combination of empirical sequencing and retrieval of unannotated Genome Survey Sequence database entries was successfully used to construct a full-length representative of the Diaspora family in Glycine max. Diaspora is presently the only fully characterized member of a lineage of putative plant endogenous retroviruses that contains virtually no trace of an extra coding region. The loss of an envelope-like coding domain suggests that non-infectious retrotransposons could swiftly evolve from infectious retroviruses, possibly by anomalous splicing of genomic RNA. PMID- 15876352 TI - A controlled trial of automated classification of negation from clinical notes. AB - BACKGROUND: Identification of negation in electronic health records is essential if we are to understand the computable meaning of the records: Our objective is to compare the accuracy of an automated mechanism for assignment of Negation to clinical concepts within a compositional expression with Human Assigned Negation. Also to perform a failure analysis to identify the causes of poorly identified negation (i.e. Missed Conceptual Representation, Inaccurate Conceptual Representation, Missed Negation, Inaccurate identification of Negation). METHODS: 41 Clinical Documents (Medical Evaluations; sometimes outside of Mayo these are referred to as History and Physical Examinations) were parsed using the Mayo Vocabulary Server Parsing Engine. SNOMED-C was used to provide concept coverage for the clinical concepts in the record. These records resulted in identification of Concepts and textual clues to Negation. These records were reviewed by an independent medical terminologist, and the results were tallied in a spreadsheet. Where questions on the review arose Internal Medicine Faculty were employed to make a final determination. RESULTS: SNOMED-CT was used to provide concept coverage of the 14,792 Concepts in 41 Health Records from John's Hopkins University. Of these, 1,823 Concepts were identified as negative by Human review. The sensitivity (Recall) of the assignment of negation was 97.2% (p < 0.001, Pearson Chi-Square test; when compared to a coin flip). The specificity of assignment of negation was 98.8%. The positive likelihood ratio of the negation was 81. The positive predictive value (Precision) was 91.2% CONCLUSION: Automated assignment of negation to concepts identified in health records based on review of the text is feasible and practical. Lexical assignment of negation is a good test of true Negativity as judged by the high sensitivity, specificity and positive likelihood ratio of the test. SNOMED-CT had overall coverage of 88.7% of the concepts being negated. PMID- 15876353 TI - Stimulation of Myc transactivation by the TATA binding protein in promoter reporter assays. AB - BACKGROUND: The c-Myc oncogenic transcription factor heterodimerizes with Max, binds specific DNA sites and regulates transcription. The role of Myc in transcriptional activation involves its binding to TRRAP and histone acetylases; however, Myc's ability to activate transcription in transient transfection assays is remarkably weak (2 to 5 fold) when compared to other transcription factors. Since a deletion Myc mutant D106-143 and a substitution mutant W135E that weakly binds TRRAP are still fully active in transient transfection reporter assays and the TATA binding protein (TBP) has been reported to directly bind Myc, we sought to determine the effect of TBP on Myc transactivation. RESULTS: We report here a potent stimulation of Myc transactivation by TBP, allowing up to 35-fold transactivation of reporter constructs. Although promoters with an initiator (InR) element briskly responded to Myc transactivation, the presence of an InR significantly diminished the response to increasing amounts of TBP. We surmise from these findings that promoters containing both TATA and InR elements may control Myc responsive genes that require brisk increased expression within a narrow window of Myc levels, independent of TBP. In contrast, promoters driven by the TATA element only, may also respond to modulation of TBP activity or levels. CONCLUSION: Our observations not only demonstrate that TBP is limiting for Myc transactivation in transient transfection experiments, but they also suggest that the inclusion of TBP in Myc transactivation assays may further improve the characterization of c-Myc target genes. PMID- 15876354 TI - Gene expression profile of cervical tissue compared to exfoliated cells: impact on biomarker discovery. AB - BACKGROUND: Exfoliated cervical cells are used in cytology-based cancer screening and may also be a source for molecular biomarkers indicative of neoplastic changes in the underlying tissue. However, because of keratinization and terminal differentiation it is not clear that these cells have an mRNA profile representative of cervical tissue, and that the profile can distinguish the lesions targeted for early detection. RESULTS: We used whole genome microarrays (25,353 unique genes) to compare the transcription profiles from seven samples of normal exfoliated cells and one cervical tissue. We detected 10,158 genes in exfoliated cells, 14,544 in the tissue and 7320 genes in both samples. For both sample types the genes grouped into the same major gene ontology (GO) categories in the same order, with exfoliated cells, having on average 20% fewer genes in each category. We also compared microarray results of samples from women with cervical intraepithelial neoplasia grade 3 (CIN3, n = 15) to those from age and race matched women without significant abnormalities (CIN1, CIN0; n = 15). We used three microarray-adapted statistical packages to identify differential gene expression. The six genes identified in common were two to four fold upregulated in CIN3 samples. One of these genes, the ubiquitin-conjugating enzyme E2 variant 1, participates in the degradation of p53 through interaction with the oncogenic HPV E6 protein. CONCLUSION: The findings encourage further exploration of gene expression using exfoliated cells to identify and validate applicable biomarkers. We conclude that the gene expression profile of exfoliated cervical cells partially represents that of tissue and is complex enough to provide potential differentiation between disease and non-disease. PMID- 15876355 TI - Three microarray platforms: an analysis of their concordance in profiling gene expression. AB - BACKGROUND: Microarrays for the analysis of gene expression are of three different types: short oligonucleotide (25-30 base), long oligonucleotide (50-80 base), and cDNA (highly variable in length). The short oligonucleotide and cDNA arrays have been the mainstay of expression analysis to date, but long oligonucleotide platforms are gaining in popularity and will probably replace cDNA arrays. As part of a validation study for the long oligonucleotide arrays, we compared and contrasted expression profiles from the three formats, testing RNA from six different cell lines against a universal reference standard. RESULTS: The three platforms had 6430 genes in common. In general, correlation of gene expression levels across the platforms was good when defined by concordance in the direction of expression difference (upregulation or downregulation), scatter plot analysis, principal component analysis, cell line correlation or quantitative RT-PCR. The overall correlations (r values) between platforms were in the range 0.7 to 0.8, as determined by analysis of scatter plots. When concordance was measured for expression ratios significant at p-values of <0.05 and at expression threshold levels of 1.5 and 2-fold, the agreement among the platforms was very high, ranging from 93% to 100%. CONCLUSION: Our results indicate that the long oligonucleotide platform is highly suitable for expression analysis and compares favorably with the cDNA and short oligonucleotide varieties. All three platforms can give similar and reproducible results if the criterion is the direction of change in gene expression and minimal emphasis is placed on the magnitude of change. PMID- 15876356 TI - Ataxia and peripheral nerve hypomyelination in ADAM22-deficient mice. AB - BACKGROUND: ADAM22 is a member of the ADAM gene family, but the fact that it is expressed only in the nervous systems makes it unique. ADAM22's sequence similarity to other ADAMs suggests it to be an integrin binder and thus to have a role in cell-cell or cell-matrix interactions. To elucidate the physiological functions of ADAM22, we employed gene targeting to generate ADAM22 knockout mice. RESULTS: ADAM22-deficient mice were produced in a good accordance with the Mendelian ratio and appeared normal at birth. After one week, severe ataxia was observed, and all homozygotes died before weaning, probably due to convulsions. No major histological abnormalities were detected in the cerebral cortex or cerebellum of the homozygous mutants; however, marked hypomyelination of the peripheral nerves was observed. CONCLUSION: The results of our study demonstrate that ADAM22 is closely involved in the correct functioning of the nervous system. Further analysis of ADAM22 will provide clues to understanding the mechanisms of human diseases such as epileptic seizures and peripheral neuropathy. PMID- 15876357 TI - Rational polynomial representation of ribonucleotide reductase activity. AB - BACKGROUND: Recent data suggest that ribonucleotide reductase (RNR) exists not only as a heterodimer R12R22 of R12 and R22 homodimers, but also as tetramers R14R24 and hexamers R16R26. Recent data also suggest that ATP binds the R1 subunit at a previously undescribed hexamerization site, in addition to its binding to previously described dimerization and tetramerization sites. Thus, the current view is that R1 has four NDP substrate binding possibilities, four dimerization site binding possibilities (dATP, ATP, dGTP, or dTTP), two tetramerization site binding possibilities (dATP or ATP), and one hexamerization site binding possibility (ATP), in addition to possibilities of unbound site states. This large number of internal R1 states implies an even larger number of quaternary states. A mathematical model of RNR activity which explicitly represents the states of R1 currently exists, but it is complicated in several ways: (1) it includes up to six-fold nested sums; (2) it uses different mathematical structures under different substrate-modulator conditions; and (3) it requires root solutions of high order polynomials to determine R1 proportions in mono-, di-, tetra- and hexamer states and thus RNR activity as a function of modulator and total R1 concentrations. RESULTS: We present four (one for each NDP) rational polynomial models of RNR activity as a function of substrate and reaction rate modifier concentrations. The new models avoid the complications of the earlier model without compromising curve fits to recent data. CONCLUSION: Compared to the earlier model of recent data, the new rational polynomial models are simpler, adequately fitting, and likely better suited for biochemical network simulations. PMID- 15876358 TI - Elevated expression of CD30 in adult T-cell leukemia cell lines: possible role in constitutive NF-kappaB activation. AB - BACKGROUND: Human T-cell leukemia virus type 1 (HTLV-1) is associated with the development of adult T-cell leukemia (ATL). HTLV-1 encoded Tax1 oncoprotein activates the transcription of genes involved in cell growth and anti-apoptosis through the NF-kappaB pathway, and is thought to play a critical role in the pathogenesis of ATL. While Tax1 expression is usually lost or minimal in ATL cells, these cells still show high constitutive NF-kappaB activity, indicating that genetic or epigenetic changes in ATL cells induce activation independent of Tax1. The aim of this study was to identify the molecules responsible for the constitutive activation of NF-kappaB in ATL cells using a retroviral functional cloning strategy. RESULTS: Using enhanced green fluorescent protein (EGFP) expression and blasticidin-resistance as selection markers, several retroviral cDNA clones exhibiting constitutive NF-kappaB activity in Rat-1 cells, including full-length CD30, were obtained from an ATL cell line. Exogenous stable expression of CD30 in Rat-1 cells constitutively activated NF-kappaB. Elevated expression of CD30 was identified in all ATL lines examined, and primary ATL cells from a small number of patients (8 out of 66 cases). CONCLUSION: Elevated CD30 expression is considered one of the causes of constitutive NF-kappaB activation in ATL cells, and may be involved in ATL development. PMID- 15876359 TI - Cellular mechanisms underlying the effects of an early experience on cognitive abilities and affective states. AB - : In the present study we investigated the effects of neonatal handling, an animal model of early experience, on spatial learning and memory, on hippocampal glucocorticoid (GR), mineralocorticoid (MR) and type 1A serotonin (5-HT1A) receptors, as well as brain derived neurotrophic factor (BDNF), and on circulating leptin levels, of male rats. METHOD: Spatial learning and memory following an acute restraint stress (30 min) were assessed in the Morris water maze. Hippocampal GR, MR and BDNF levels were determined immunocytochemically. 5 HT1A receptors were quantified by in vitro binding autoradiography. Circulating leptin levels, following a chronic forced swimming stress, were measured by radioimmunoassay (RIA). Data were statistically analyzed by analysis of variance (ANOVA). RESULTS: Neonatal handling increased the ability of male rats for spatial learning and memory. It also resulted in increased GR/MR ratio, BDNF and 5-HT1A receptor levels in the hippocampus. Furthermore, leptin levels, body weight and food consumption during chronic forced swimming stress were reduced as a result of handling. CONCLUSION: Neonatal handling is shown to have a beneficial effect in the males, improving their cognitive abilities. This effect on behavior could be mediated by the handling-induced increase in hippocampal GR/MR ratio and BDNF levels. The handling-induced changes in BDNF and 5-HT1A receptors could underlie the previously documented effect of handling in preventing "depression". Furthermore, handling is shown to prevent other maladaptive states such as stress induced hyperphagia, obesity and resistance to leptin. PMID- 15876360 TI - Characteristics of patients with organic brain syndromes : A cross-sectional 2 year follow-up study in Kuala Lumpur, Malaysia. AB - BACKGROUND: Organic Brain Syndromes (OBS) are often missed in clinical practice. Determining their varied presentations may help in earlier detection, better management, and, assessing prognosis and outcome. We described the in-patient referrals of patients suffering from the psychiatric effects of organic states and compared the symptomatology and mortality between those with the Acute and Chronic varieties. METHODS: 59 patients referred to our Consultation-Liaison (C L) Psychiatry services and given a clinical diagnosis of OBS were selected over a 6-month period. Psychiatric and cognitive abnormalities and treatment regimes were recorded and fatality rates determined. Information regarding their condition 24 months after their index hospitalization was recorded. All data were entered into a proforma and analyzed after exclusion. RESULTS: The mean duration of detecting the symptoms by the physician was 3.52 days. The presence of a premorbid psychiatric illness had no influence on the clinical presentation but did on the mortality of patients with OBS (p = 0.029).Patients with the Acute syndrome had significantly more symptom resolution as compared to those with the Chronic syndrome (p = 0.001) but mortalityrates did not differ. Elderly patients and those with symptom resolution upon discharge did not show statistically significant higher mortality rates. The most popular combination of treatment was that of a low-dose neuroleptic and a benzodiazepine (34.7%). The need for maintenance treatment was not significantly different in any group, even in those with a past history of a functional disorder. CONCLUSION: Other than the Acute group having a significantly better outcome in terms of symptom resolution, our findings suggest that there was no significant difference in the clinical presentation between those with Acute or Chronic OBS. Mortality-wise, there was also no difference between the Acute and Chronic syndromes, nor was there any difference between the elderly and the younger group. There was also no significant difference in the need for continued treatment in both groups. PMID- 15876361 TI - Inflammation and atherosclerosis in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) associates with increased cardiovascular mortality. This appears to be predominantly due to ischaemic causes, such as myocardial infarction and congestive heart failure. The higher prevalence of cardiac ischaemia in RA is thought to be due to the accelerated development of atherosclerosis. There are two main reasons for this, which might be inter related: the systemic inflammatory load, characteristic of RA; and the accumulation in RA of classical risk factors for coronary heart disease, which is reminiscent of the metabolic syndrome. We describe and discuss in the context of RA the involvement of local and systemic inflammatory processes in the development and rupture of atherosclerotic plaques, as well as the role of individual risk factors for coronary heart disease. We also present the challenges facing the clinical and scientific communities addressing this problem, which is receiving increasing attention. PMID- 15876362 TI - A randomised controlled trial to compare the cost-effectiveness of tricyclic antidepressants, selective serotonin reuptake inhibitors and lofepramine. AB - OBJECTIVE: To determine the relative cost-effectiveness of three classes of antidepressants: tricyclic antidepressants (TCAs), selective serotonin reuptake inhibitors (SSRIs), and the modified TCA lofepramine, as first choice treatments for depression in primary care. DESIGN: Open, pragmatic, controlled trial with three randomised arms and one preference arm. Patients were followed up for 12 months. SETTING: UK primary care: 73 practices in urban and rural areas in England. PARTICIPANTS: Patients with a new episode of depressive illness according to GP diagnosis. INTERVENTIONS: Patients were randomised to receive a TCA (amitriptyline, dothiepin or imipramine), an SSRI (fluoxetine, sertraline or paroxetine) or lofepramine. Patients or GPs were able to choose an alternative treatment if preferred. MAIN OUTCOME MEASURES: At baseline the Clinical Interview Schedule, Revised (CIS-R PROQSY computerised version) was administered to establish symptom profiles. Outcome measures over the 12-month follow-up included the Hospital Anxiety and Depression Scale self-rating of depression (HAD-D), CIS R, EuroQol (EQ-5D) for quality of life, Short Form (SF-36) for generic health status, and patient and practice records of use of health and social services. The primary effectiveness outcome was the number of depression-free weeks (HAD-D less than 8, with interpolation of intervening values) and the primary cost outcome total direct NHS costs. Quality-adjusted life-years (QALYs) were used as the outcome measure in a secondary analysis. Incremental cost-effectiveness ratios and cost-effectiveness acceptability curves were computed. Estimates were bootstrapped with 5000 replications. RESULTS: In total, 327 patients were randomised. Follow-up rates were 68% at 3 months and 52% at 1 year. Linear regression analysis revealed no significant differences between groups in number of depression-free weeks when adjusted for baseline HAD-D. A higher proportion of patients randomised to TCAs entered the preference arm than those allocated to the other choices. Switching to another class of antidepressant in the first few weeks of treatment occurred significantly more often in the lofepramine arm and less in the preference arm. There were no significant differences between arms in mean cost per depression-free week. For values placed on an additional QALY of over 5000 pounds, treatment with SSRIs was likely to be the most cost-effective strategy. TCAs were the least likely to be cost-effective as first choice of antidepressant for most values of a depression-free week or QALY respectively, but these differences were relatively modest. CONCLUSIONS: When comparing the different treatment options, no significant differences were found in outcomes or costs within the sample, but when outcomes and costs were analysed together, the resulting cost-effectiveness acceptability curves suggested that SSRIs were likely to be the most cost-effective option, although the probability of this did not rise above 0.6. Choosing lofepramine is likely to lead to a greater proportion of patients switching treatment in the first few weeks. Further research is still needed on the management of depressive illness in primary care. This should address areas such as the optimum severity threshold at which medication should be used; the feasibility and effectiveness of adopting structured depression management programmes in the UK context; the importance of factors such as physical co-morbidity and recent life events in GPs' prescribing decisions; alternative ways of collecting data; and the factors that give rise to many patients being reluctant to accept medication and discontinue treatment early. PMID- 15876363 TI - Clinical effectiveness and cost-effectiveness of immediate angioplasty for acute myocardial infarction: systematic review and economic evaluation. AB - OBJECTIVES: To review the clinical evidence comparing immediate angioplasty with thrombolysis, and to consider whether it would be cost-effective. DATA SOURCES: Electronic databases. Experts in the field. REVIEW METHODS: For clinical effectiveness, a comprehensive review of randomised control trials (RCTs) was used for efficacy, and a selection of observational studies such as case series or audit data used for effectiveness in routine practice. RCTs of thrombolysis were used to assess the relative value of prehospital and hospital thrombolysis. Observational studies were used to assess the representativeness of patients in the RCTs, and to determine whether different groups have different capacity to benefit. Clinical effectiveness was synthesised through a narrative review with full tabulation of results of all included studies and a meta-analysis to provide a precise estimate of absolute clinical benefit. Consideration was given to the effect of the growing use of stents. The economic modelling adopted an NHS perspective to develop a decision-analytical model of cost-effectiveness focusing on opportunity costs over the short term (6 months). RESULTS: The results were consistent in showing an advantage of immediate angioplasty over hospital thrombolysis. The updated meta-analysis showed that mortality is reduced by about one-third, from 7.6% to 4.9% in the first 6 months, and by about the same in studies of up to 24 months. Reinfarction is reduced by over half, from 7.6% to 3.1%. Stroke is reduced by about two-thirds, from 2.3% with thrombolysis to 0.7% with percutaneous coronary intervention (PCI), with the difference being due to haemorrhagic stroke. The need for coronary artery bypass graft is reduced by about one-third, from 13.2% to 8.4%. Caution is needed in interpreting some of the older trials, as changes such as an increase in stenting and the use of the glycoprotein IIb/IIa inhibitors may improve the results of PCI. There is little evidence comparing prehospital thrombolysis with immediate PCI. Research on thrombolysis followed by PCI, known as 'facilitated PCI', is underway, but results are not yet available. Trials may be done in select centres and results may not be as good in lower volume centres, or out of normal working hours. In addition, much of the marginal mortality benefit of PCI over hospital thrombolysis may be lost if door-to-balloon time were more than an hour longer than door-to-needle time. Conversely, within the initial 6 hours, the later patients present, the greater the relative advantage of PCI. Results suggest that PCI is more cost-effective than thrombolysis, providing additional benefits in health status at some extra cost. In the longer term, the cost difference is expected to be reduced because of higher recurrence and reintervention rates among those who had thrombolysis. CONCLUSIONS: If both interventions were routinely available, the economic analysis favours PCI, given the assumptions of the model. However, very few units in England could offer a routine immediate PCI service at present, and there would be considerable resource implications of setting up such services. Without a detailed survey of existing provision, it is not possible to quantify the implications, but they include both capital and revenue: an increase in catheter laboratory provision and running costs. The greatest problem would be staffing, and that would take some years to resolve. A gradual incrementalist approach based on clinical networks, with transfer to centres able to offer PCI, may be used. In rural areas, one option may be to promote an increase in prehospital thrombolysis, with PCI for thrombolysis failures. There is a need for data on the long-term consequences of treatment, the quality of life of patients after treatment, and the effects of PCI following thrombolysis failure. PMID- 15876364 TI - Structure of a cyanobacterial BLUF protein, Tll0078, containing a novel FAD binding blue light sensor domain. AB - The sensor proteins for blue light using the FAD (BLUF) domain belong to the third family of the photoreceptor proteins using a flavin chromophore, where the other two families are phototropins and cryptochromes. As the first structure of this BLUF domain, we have determined the crystal structure of the Tll0078 protein from Thermosynechococcus elongatus BP-1, which contains a BLUF domain bound to FAD, at 2A resolution. Five Tll0078 monomers are located around the non crystallographic 5-fold axis to form a pentamer, and two pentamers related by 2 fold non-crystallographic symmetry form a decameric assembly. The monomer consists of two domains, the BLUF domain at the N-terminal region and the C terminal domain. The overall structure of the BLUF domain consists of a five stranded mixed beta-sheet with two alpha-helices running parallel with it. The isoalloxazine ring of FAD is accommodated in a pocket formed by several highly conserved amino acid residues in the BLUF domain. Of these, the three apparent key residues (Asn31, Asn32 and Gln50) were substituted with Ala. Mutant proteins of N31A and N32A showed a nearly normal 10nm spectral shift of the flavin upon illumination, while the Q50A mutant did not exhibit such a shift at all. On the basis of the crystal structure, we discussed a possible role of Gln50, which is structurally and functionally linked with the critical Tyr8 (FAD-Gln50-Tyr8 network), with regard to the light-induced spectral shift of the BLUF proteins. PMID- 15876365 TI - Structural analysis and solution studies of the activated regulatory domain of the response regulator ArcA: a symmetric dimer mediated by the alpha4-beta5 alpha5 face. AB - Escherichia coli react to changes from aerobic to anaerobic conditions of growth using the ArcA-ArcB two-component signal transduction system. This system, in conjunction with other proteins, regulates the respiratory metabolic pathways in the organism. ArcA is a member of the OmpR/PhoB subfamily of response regulator transcription factors that are known to regulate transcription by binding in tandem to target DNA direct repeats. It is still unclear in this subfamily how activation by phosphorylation of the regulatory domain of response regulators stimulates DNA binding by the effector domain and how dimerization and domain orientation, as well as intra- and intermolecular interactions, affect this process. In order to address these questions we have solved the crystal structures of the regulatory domain of ArcA in the presence and absence of the phosphoryl analog, BeF3-. In the crystal structures, the regulatory domain of ArcA forms a symmetric dimer mediated by the alpha4-beta5-alpha5 face of the protein and involving a number of residues that are highly conserved in the OmpR/PhoB subfamily. It is hypothesized that members of this subfamily use a common mechanism of regulation by dimerization. Additional biophysical studies were employed to probe the oligomerization state of ArcA, as well as its individual domains, in solution. The solution studies show the propensity of the individual domains to associate into oligomers larger than the dimer observed for the intact protein, and suggest that the C-terminal DNA-binding domain also plays a role in oligomerization. PMID- 15876366 TI - Integrated pseudogene annotation for human chromosome 22: evidence for transcription. AB - Pseudogenes are inheritable genetic elements formally defined by two properties: their similarity to functioning genes and their presumed lack of activity. However, their precise characterization, particularly with respect to the latter quality, has proven elusive. An opportunity to explore this issue arises from the recent emergence of tiling-microarray data showing that intergenic regions (containing pseudogenes) are transcribed to a great degree. Here we focus on the transcriptional activity of pseudogenes on human chromosome 22. First, we integrated several sets of annotation to define a unified list of 525 pseudogenes on the chromosome. To characterize these further, we developed a comprehensive list of genomic features based on conservation in related organisms, expression evidence, and the presence of upstream regulatory sites. Of the 525 unified pseudogenes we could confidently classify 154 as processed and 49 as duplicated. Using data from tiling microarrays, especially from recent high-resolution oligonucleotide arrays, we found some evidence that up to a fifth of the 525 pseudogenes are potentially transcribed. Expressed sequence tags (EST) comparison further validated a number of these, and overall we found 17 pseudogenes with strong support for transcription. In particular, one of the pseudogenes with both EST and microarray evidence for transcription turned out to be a duplicated pseudogene in the cat eye syndrome critical region. Although we could not identify a meaningful number of transcription factor-binding sites (based on chromatin immunoprecipitation-chip data) near pseudogenes, we did find that approximately 12% of the pseudogenes had upstream CpG islands. Finally, analysis of corresponding syntenic regions in the mouse, rat and chimp genomes indicates, as previously suggested, that pseudogenes are less conserved than genes, but more preserved than the intergenic background (all notation is available from http://www.pseudogene.org). PMID- 15876367 TI - Multiple effects of S13 in modulating the strength of intersubunit interactions in the ribosome during translation. AB - The ribosomal protein S13 is found in the head region of the small subunit, where it interacts with the central protuberance of the large ribosomal subunit and with the P site-bound tRNA through its extended C terminus. The bridging interactions between the large and small subunits are dynamic, and are thought to be critical in orchestrating the molecular motions of the translation cycle. S13 provides a direct link between the tRNA-binding site and the movements in the head of the small subunit seen during translocation, thereby providing a possible pathway of signal transduction. We have created and characterized an rpsM(S13) deficient strain of Escherichia coli and have found significant defects in subunit association, initiation and translocation through in vitro assays of S13 deficient ribosomes. Targeted mutagenesis of specific bridge and tRNA contact elements in S13 provides evidence that these two interaction domains play critical roles in maintaining the fidelity of translation. This ribosomal protein thus appears to play a non-essential, yet important role by modulating subunit interactions in multiple steps of the translation cycle. PMID- 15876368 TI - Regulation by oligomerization in a mycobacterial folate biosynthetic enzyme. AB - Folate derivatives are essential cofactors in the biosynthesis of purines, pyrimidines and amino acids across all forms of life. Mammals uptake folate from their diets, whereas most bacteria must synthesize folate de novo. Therefore, the enzymes in the folate biosynthetic pathway are attractive drug targets against bacterial pathogens such as Mycobacterium tuberculosis, the cause of the world's most deadly infectious disease, tuberculosis (TB). M.tuberculosis 7,8 dihydroneopterin aldolase (Mtb FolB, DHNA) is the second enzyme in the folate biosynthetic pathway, which catalyzes the conversion of 7,8-dihydroneopterin to 6 hydroxymethyl-7,8-dihydropterin and glycoaldehyde. The 1.6A X-ray crystal structure of Mtb FolB complexed with its product, 6-hydroxymethyl-7,8 dihydropterin, reveals an octameric assembly similar to that seen in crystal structures of other FolB homologs. However, the 2.5A crystal structure of unliganded Mtb FolB reveals a novel tetrameric oligomerization state, with only partially formed active sites. A substrate induced conformational change appears to be necessary to convert the inactive tetramer to the active octamer. Ultracentrifugation confirmed that in solution unliganded Mtb FolB is mainly tetrameric and upon addition of substrate FolB is predominantly octameric. Kinetic analysis of substrate binding gives a Hill coefficient of 2.0, indicating positive cooperativity. We hypothesize that Mtb FolB displays cooperativity in substrate binding to regulate the cellular concentration of 7,8-dihydroneopterin, so that it may function not only as a precursor to folate but also as an antioxidant for the survival of M.tuberculosis against host defenses. PMID- 15876369 TI - Evidence for the intramolecular pleating model of fibrillin microfibril organisation from single particle image analysis. AB - Fibrillin microfibrils endow mammalian connective tissues with elasticity and are fundamental for the deposition of elastin. The microfibrils are 57nm periodic supramolecular protein polymers with a mass of 2.4MDa per repeat. The detailed structure and organisation of most matrix assemblies is poorly understood due to their large size and complexity and it has proved a major challenge to define their structural organisation. Therefore, we have used low dose electron microscopy and single particle image analysis to study the structure of fibrillin microfibrils. Three novel features were detected: a globular feature that bridges the "arm" region, a double band of density crossing the microfibril and stain penetrating holes present in the interbead region, possibly produced by the removal of microfibril associated proteins in the purification procedure. Fine filaments of approximately 2.4nm diameter are resolved in the interbead region, which correspond to the reported diameter of the fibrillin molecule. Comparison of the stain exclusion pattern of microfibrils with the theoretical stain exclusion pattern of fibrillin packing models indicates that the intramolecular pleating model, where each fibrillin molecule is pleated within one microfibril period allowing extensibility by unpleating, has the best fit to the data. PMID- 15876370 TI - Effect of inorganic phosphate on FMN binding and loop flexibility in Desulfovibrio desulfuricans apo-flavodoxin. AB - The complex between flavin mononucleotide (FMN) and apo-flavodoxin is dominated by isoalloxazine-stacking interactions and 5'-phosphate hydrogen bonds. We show here that FMN binding to Desulfovibrio desulfuricans apo-flavodoxin is faster and the affinity is higher in the presence of inorganic phosphate as compared to in its absence (I=110 mM, pH 7, 20 degrees C). The transition-state of complex formation was investigated by phi-value analysis using Trp60Ala and Tyr98Ala apo flavodoxin variants. We find that Tyr98 is highly involved in the FMN/protein transition state independent of inorganic phosphate, whereas the participation of Trp60 is modulated by inorganic phosphate. The phi-value for Trp60 is higher without phosphate, implying that at this condition stronger binding of Trp60 is required in the transition state to assure successful complex formation. Consistent with the experimental data, all-atom molecular dynamic simulations reveal that the presence of an anion in the phosphate subsite restricts the mobility of the Trp60-containing loop in terms of both backbone and side-chain movements, but has no effect on the Tyr98-containing loop. The overall thermodynamic stability of apo-flavodoxin is higher in the presence of inorganic phosphate as compared to in its absence (I=110 mM, pH 7, 20 degrees C). Kinetic experiments reveal that the additional stability originates in slower unfolding. The combined experimental and computational observations demonstrate that phosphate has an ordering effect on the Trp60-containing loop, which positions Trp60 favorably for FMN binding and increases the barrier for protein unfolding. PMID- 15876371 TI - Crystal structure of the E. coli dipeptidyl carboxypeptidase Dcp: further indication of a ligand-dependent hinge movement mechanism. AB - Dcp from Escherichia coli is a 680 residue cytoplasmic peptidase, which shows a strict dipeptidyl carboxypeptidase activity. Although Dcp had been assigned to the angiotensin I-converting enzymes (ACE) due to blockage by typical ACE inhibitors, it is currently grouped into the M3 family of mono zinc peptidases, which also contains the endopeptidases neurolysin and thimet oligopeptidase (TOP). We have cloned, expressed, purified, and crystallized Dcp in the presence of an octapeptide "inhibitor", and have determined its 2.0A crystal structure using MAD methods. The analysis revealed that Dcp consists of two half shell-like subdomains, which enclose an almost closed two-chamber cavity. In this cavity, two dipeptide products presumably generated by Dcp cleavage of the octapeptide bind to the thermolysin-like active site fixed to side-chains, which are provided by both subdomains. In particular, an Arg side-chain backed by a Glu residue, together with two Tyr phenolic groups provide a charged anchor for fixing the C terminal carboxylate group of the P2' residue of a bound substrate, explaining the strict dipeptidyl carboxypeptidase specificity of Dcp. Tetrapeptidic substrates are fixed only via their main-chain functions from P2 to P2', suggesting a broad residue specificity for Dcp. Both subdomains exhibit very similar chain folds as the equivalent but abducted subdomains of neurolysin and TOP. Therefore, this "product-bound" Dcp structure seems to represent the inhibitor/substrate-bound "closed" form of the M3 peptidases, generated from the free "open" substrate-accessible form by a hinge-bending mechanism. A similar mechanism has recently been demonstrated experimentally for ACE2. PMID- 15876372 TI - Malaria parasite actin filaments are very short. AB - A novel form of actomyosin regulation has recently been proposed in which the polymerisation of new actin filaments regulates apicomplexan parasite motility. Here, we identified actin I in the merozoites of Plasmodium falciparum by mass spectrometry. The only post-translational modification is acetylation of the N terminus (acetyl-Gly-Glu-actin), while methylation of histidine 73, a common modification for actin, is absent. Results obtained with anti-actin antibodies suggest that, in contrast to a previous report, there is no actin-ubiquitin conjugate in merozoites. About half of the extracted monomeric actin polymerised and actin filaments could be sedimented at 500,000g. In contrast, centrifugation at 100,000g, conditions commonly used to sediment filamentous actin, yielded very little F-actin. In a functional characterisation using an in vitro motility assay, actin filaments moved over myosin at a velocity indistinguishable from that of rabbit skeletal actin. Filament length, however, was too short to be resolved by conventional fluorescence microscopy. On electron micrographs an average filament length of approximately 100nm was determined. We also identified by mass spectrometry proteins co-purifying with filamentous actin, which are potential actin-binding proteins. Our results demonstrate differences in actin filament dynamics for an apicomplexan parasite, which could be due to specific properties of the actin and/or actin-regulatory proteins. PMID- 15876373 TI - Understanding carbamoyl phosphate synthetase deficiency: impact of clinical mutations on enzyme functionality. AB - Carbamoyl phosphate synthetase I (CPSI) deficiency, a recessively inherited error of the urea cycle, causes life-threatening hyperammonaemia. CPSI is a multidomain 1500-residue liver mitochondrial matrix protein that is allosterically activated by N-acetyl-l-glutamate, and which synthesises carbamoyl phosphate (CP) in three steps: bicarbonate phosphorylation by ATP, carbamate synthesis from carboxyphosphate and ammonia, and carbamate phosphorylation by ATP. Several missense mutations of CPSI have been reported in patients with CPSI deficiency, but the actual pathogenic potential and effects on the enzyme of these mutations remain non-characterised. Since the structure of Escherichia coli CPS is known and systems for its overexpression and purification are available, we have constructed and purified eight site-directed mutants of E.coli CPS affecting the enzyme large subunit (A126M, R169H, Q262P, N301K, P360L, V640R, R675L, S789P) that are homologous to corresponding missense mutations found in patients with CPSI deficiency, studying their stability and their ability to catalyse the CPS reaction as well as the partial reactions that reflect the different reactional steps, and analysing the substrate kinetics for the overall and partial reactions. The results show that all the mutations significantly decrease CP synthesis without completely inactivating the enzyme (as reflected in the catalysis of at least one partial reaction), that one of these mutations (Q262P) causes marked enzyme instability, and validate the use of E.coli CPS as a pathogenicity testing model for CPSI deficiency. The causality of the reported clinical mutations is supported and the derangements caused by the mutations are identified, revealing the specific roles of the residues that are mutated. In particular, the findings highlight the importance for carbamate phosphorylation and for allosteric activation of a loop that coordinates K(+), stress the key role of intersubunit interactions for CPS stability, and suggest that lid opening at both phosphorylation sites is concerted. PMID- 15876374 TI - Structural stability of soybean lipoxygenase-1 in solution as probed by small angle X-ray scattering. AB - Soybean lipoxygenase-1 (LOX-1) is used widely as a model for studying the structural and functional properties of the homologous family of lipoxygenases. The crystallographic structure revealed that LOX-1 is organized in a beta-sheet N terminal domain and a larger, mostly helical, C-terminal domain. Here, we describe the overall structural characterization of native unliganded LOX-1 in solution, using small angle X-ray scattering (SAXS). We show that the scattering pattern of the unliganded enzyme in solution does not display any significant difference compared with that calculated from the crystal structure, and that models of the overall shape of the protein calculated ab initio from the SAXS pattern provide a close envelope to the crystal structure. These data, demonstrating that LOX-1 has a compact structure also in solution, rule out any major motional flexibility of the LOX-1 molecule in aqueous solutions. In addition we show that eicosatetraynoic acid, an irreversible inhibitor of lipoxygenase used to mimic the effect of substrate binding, does not alter the overall conformation of LOX-1 nor its ability to bind to membranes. In contrast, the addition of glycerol (to 5%, v/v) causes an increase in the binding of the enzyme to membranes without altering its catalytic efficiency towards linoleic acid nor its SAXS pattern, suggesting that the global conformation of the enzyme is unaffected. Therefore, the compact structure determined in the crystal appears to be essentially preserved in these various solution conditions. During the preparation of this article, a paper by M. Hammel and co-workers showed instead a sharp difference between crystal and solution conformations of rabbit 15-LOX-1. The possible cause of this difference might be the presence of oligomers in the rabbit lipoxygenase preparations. PMID- 15876375 TI - Structural insights into the monosaccharide specificity of Escherichia coli rhamnose mutarotase. AB - The crystal structure of Escherichia coli rhamnose mutarotase (YiiL) is completely different from the previously reported structures of the Lactococcus lactis galactose mutarotase and the Bacillus subtilis RbsD (pyranase). YiiL exists as a locally asymmetric dimer, which is stabilized by an intermolecular beta-sheet, various hydrophobic interactions, and a cation-pi interaction with a salt-bridge. The protein folds of YiiL are similar to those of a Streptomyces coelicolor mono-oxygenase and a hypothetical Arabidopsis thaliana protein At3g17210. By assaying the enzymatic activity of six active-site mutants and by comparing the crystal structure-derived active site conformations of YiiL, RbsD, and a galactose mutarotase, we were able to define the amino acid residues required for catalysis and suggest a possible catalytic mechanism for YiiL. Although the active-site amino acid residues of YiiL (His, Tyr, and Trp) differ greatly from those of galactose mutarotase (His, Glu, and Asp), their geometries, which determine the structures of the preferred monosaccharide substrates, are conserved. In addition, the in vivo function of YiiL was assessed by constructing a mutant E.coli strain that carries a yiiL deletion. The presence of the yiiL gene is critical for efficient cell growth only when concentrations of l-rhamnose are limited. PMID- 15876376 TI - NMR investigations of allosteric processes in a two-domain Thermus thermophilus Hsp70 molecular chaperone. AB - Hsp70 chaperones are two-domain proteins that assist in intra-cellular protein (re) folding processes in all species. The protein folding activity of the substrate binding domain of the Hsp70s is regulated by nucleotide binding at the nucleotide-binding domain through an as yet undefined heterotropic allosteric mechanism. The available structures of the isolated domains of Hsp70s have given very limited indications of nucleotide-induced conformational changes that could modulate the affinity for substrate proteins. Here, we present a multi dimensional NMR study of a prokaryotic Hsp70 homolog, Thermus thermophilus DnaK, using a 54kDa construct containing both nucleotide binding domain and most of the substrate binding domain. It is determined that the nucleotide binding domain and substrate binding domain are closely associated in all ligand states studied. Comparison of the assigned NMR spectra of the two-domain construct with those of the previously studied isolated nucleotide binding domain, allowed the identification of the nucleotide binding domain-substrate binding domain interface. A global three-dimensional structure was obtained for the two-domain construct on the basis of this information and of NMR residual dipolar couplings measurements. This is the first experimental elucidation of the relative positioning of the nucleotide binding domain and substrate binding domain for any Hsp70 chaperone. Comparisons of NMR data between various ligand states including nucleotide-free, ATP, ADP.Pi and ADP.Pi+ peptide bound, identified residues involved in the allosteric inter-domain communication. In particular, peptide binding to the substrate binding domain was found to cause conformational changes in the NBD extending to the nucleotide binding pocket. Detailed analysis suggests that the inter-domain interface becomes tighter in the (nucleotide binding domain ligation/substrate binding domain ligation) order ATP/apo, ADP.Pi/apo ADP.Pi/peptide. PMID- 15876377 TI - What contributions to protein side-chain dynamics are probed by NMR experiments? A molecular dynamics simulation analysis. AB - Molecular dynamics simulations of the structurally homologous proteins TNfn3 and FNfn10 have been used to investigate the contributions to side-chain dynamics measured by NMR relaxation experiments. The results reproduce the variation in core side-chain dynamics observed by NMR and highlight the relevance of anharmonic motion and transitions between local minima for explaining NMR side chain order parameters. A method is described for calculating converged order parameters by use of replica exchange molecular dynamics in conjunction with an implicit solvent model. These simulations allow the influence of various factors, such as the flexibility of side-chains and their free volume, on the mobility to be tested by perturbing the system. Deletion mutations are found to have the largest effect on the more densely packed FNfn10. Some counterintuitive effects are seen, such as an increase in order parameters close to deletion mutation sites, but these can be rationalized in terms of direct interactions with the modified side-chains. A statistical analysis of published order parameters supports the conclusions drawn from the simulations. PMID- 15876378 TI - Extending the folding nucleus of ubiquitin with an independently folding beta hairpin finger: hurdles to rapid folding arising from the stabilisation of local interactions. AB - The N-terminal beta-hairpin sequence of ubiquitin has been implicated as a folding nucleation site. To extend and stabilise the ubiquitin folding nucleus, we have inserted an autonomously folding 14-residue peptide sequence beta4 which in isolation forms a highly populated beta-hairpin (>70%) stabilised by local interactions. NMR structural analysis of the ubiquitin mutant (Ubeta4) shows that the hairpin finger is fully structured and stabilises ubiquitin by approximately 8kJmol(-1). Protein engineering and kinetic (phi(F)-value) analysis of a series of Ubeta4 mutants shows that the hairpin extension of Ubeta4 is also significantly populated in the transition state (phi(F)-values >0.7) and has the effect of templating the formation of native contacts in the folding nucleus of ubiquitin. However, at low denaturant concentrations the chevron plot of Ubeta4 shows a small deviation from linearity (roll-over effect), indicative of the population of a compact collapsed state, which appears to arise from over stabilisation of local interactions. Destabilising mutations within the native hairpin sequence and within the engineered hairpin extension, but not elsewhere, eliminate this non-linearity and restore apparent two-state behaviour. The pitfall to stabilising local interactions is to present hurdles to the rapid and efficient folding of small proteins down a smooth folding funnel by trapping partially folded or misfolded states that must unfold or rearrange before refolding. PMID- 15876379 TI - The 2.1A crystal structure of the far-red fluorescent protein HcRed: inherent conformational flexibility of the chromophore. AB - We have determined the crystal structure of HcRed, a far-red fluorescent protein isolated from Heteractis crispa, to 2.1A resolution. HcRed was observed to form a dimer, in contrast to the monomeric form of green fluorescent protein (GFP) or the tetrameric forms of the GFP-like proteins (eqFP611, Rtms5 and DsRed). Unlike the well-defined chromophore conformation observed in GFP and the GFP-like proteins, the HcRed chromophore was observed to be considerably mobile. Within the HcRed structure, the cyclic tripeptide chromophore, Glu(64)-Tyr(65)-Gly(66), was observed to adopt both a cis coplanar and a trans non-coplanar conformation. As a result of these two conformations, the hydroxyphenyl moiety of the chromophore makes distinct interactions within the interior of the beta-can. These data together with a quantum chemical model of the chromophore, suggest the cis coplanar conformation to be consistent with the fluorescent properties of HcRed, and the trans non-coplanar conformation to be consistent with non fluorescent properties of hcCP, the chromoprotein parent of HcRed. Moreover, within the GFP-like family, it appears that where conformational freedom is permissible then flexibility in the chromophore conformation is possible. PMID- 15876380 TI - Evaluation of PCDD/F levels in ambient air and soils and estimation of deposition rates in Kocaeli, Turkey. AB - PCDD/F analyses were made in soil and ambient air samples taken from Kocaeli, an industrialized area of Turkey. Results showed that the levels of PCDD/F pollution are comparable to the levels observed in the various urban areas in the world. PCDD/F concentrations in surface soils ranged between 0.4 and 4.27 pg I-TEQ kg( 1) with a geometric mean of 0.76 pg I-TEQ kg(-1), while those in ambient air were between 23 and 563 f gm(-3), with geometric mean of 82.0 f gm(-3). Ambient air PCDD/F concentration in the city center was about 10 times higher than those in the rural area. Combustion activities present in the area were assessed to be the most significant source of the PCDD/F pollution based on the congener and homologue profiles and statistical analysis of the data. The deposition rate modeling of USEPA was applied and the deposition rates were determined in the range between 7.6 and 66.3 ng I-TEQ m(-2)year(-1) with a geometric mean of 15.9 ng I-TEQ m(-2)year(-1). The rates were higher than the recommended deposition rates in relation to the tolerable daily intake (TDI) range of PCDD/Fs for humans. The deposition velocities computed were also found to be high in both low and highly chlorinated PCDD/Fs, and this was attributed to the vapor phase deposition of volatile PCDD/Fs, and the scavenging effect of the precipitation on the particle-bound PCDD/Fs. PMID- 15876381 TI - Abundance and sources of ambient dioxins in Hong Kong: a review of dioxin measurements from 1997 to 2001. AB - Ambient measurements of seventeen 2,3,7,8-polychlorinated dibenzo-p dioxin/dibenzofuran congeners (2,3,7,8-PCDD/Fs) have been taken in a number of monitoring programs or ad-hoc studies in Hong Kong. The longest monitoring program started at two locations in the territory in July 1997. The other monitoring efforts are ad-hoc studies, varying from a few coordinated sampling events at multiple sites to a year-long monitoring project that targeted suspected local dioxin sources. In this paper, we examined these measurements to understand the ambient levels, temporal and spatial variation, and possible sources of the 2,3,7,8-PCDD/Fs in Hong Kong. The territory-wide annual average concentration of the dioxins was 0.052 pg I-TEQ/m3 measured at the regular monitoring stations in the most recent annual cycle of 2000/2001. This level fell at the lower end of the range of dioxin concentrations measured at other urban locations around the world. The dioxin levels showed a clear seasonality in that elevated concentrations were observed in the winter and lower concentrations in the summer at all monitoring sites with one year or more regular measurements. The measurements indicated that the few known local dioxin sources, including a major chemical waste incinerator facility, landfill sites, and vehicular traffic, are not important contributors to ambient dioxins in Hong Kong. On days of high dioxin concentrations, the 2,3,7,8-PCDD/F congeners were observed to have almost identical compositions with a north-northwest to south-southeast spatial gradient in concentrations at different sampling locations in Hong Kong. This observation, along with other collaborative evidence, established a strong link between high dioxin concentration days in Hong Kong and regional transport of the polluted air masses from the north. PMID- 15876382 TI - Formation of PCDD/Fs in the sintering process: role of the grid-Cr2O3 catalyst in the de novo synthesis. AB - The sintering process is among the major sources of the very toxic polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) in the environment. At the industrial scale, it has been shown that dust collected on the grid, which supports the feed, contains PCDD/Fs amounts between the values found in the bottom of the cake and the values found on dust collected during gas sampling in the wind boxes. This fact suggests that the grid, containing 25wt.% of chromium, could have a catalytic activity in PCDD/Fs formation during the sintering process. This research tries to study this potential role. The de novo synthesis of PCDD/Fs is simulated at laboratory scale by thermal treatments of samples mixed with grid filings or Cr2O3. The thermal experiments performed with E.S.P. dust (dust collected in the electrostatic precipitator of a sintering plant) or graphite mixed with grid filings do not allow to confirm a role of the grid in PCDD/Fs formation during the industrial process. On the other hand, it has been shown that Cr2O3 can be considered as a catalyst in the de novo synthesis of PCDD/Fs. This compound takes place in the two steps of the de novo synthesis: the degradation of the carbon matrix as well as the chlorination reactions. PMID- 15876383 TI - Comparison of supercritical fluid extraction and Soxhlet extraction for the determination of PCBs in seaweed samples. AB - The efficiency of supercritical fluid extraction for the determination of 12 polychlorinated biphenyls from algae samples is compared to Soxhlet extraction. Analytical detection limits for the individual congeners ranged from 0.62 microgl(-1) to 19 microgl(-1). Recovery was tested for both methods using standard addition procedure. At maximum spike level of concentration, the mean recoveries were not significantly different (P>0.05) of all PCBs studied, with the exception of PCBs 28, 52, 77 and 169. Method precision for Soxhlet extraction (< or =3.9%) was slightly better than for SFE (< or =9.2%). Although both methods yield comparable results, SFE offers the advantage of detecting all PCBs studied at lower concentrations, reducing extraction time, and reducing the amount of solvents needed. The optimized methods were applied to the analysis of three real seaweed samples, except for PCB101 the concentrations of all PCBs were low or below the detection limits. The levels of PCB101 found in sample 1 were 6.6+/ 0.54 ng g(-1) d.w., in sample 2 the levels were 8.2+/-0.86 ng g(-1) d.w. and in sample 3 they were 7.7+/-0.08 ng g(-1) d.w. PMID- 15876384 TI - Anaerobic degradation of nonylphenol in sludge. AB - We investigated the effects of various factors on the anaerobic degradation of nonylphenol (NP) in sludge. NP (5 mg/l) anaerobic degradation rate constants were 0.029 1/day for sewage sludge and 0.019l/day for petrochemical sludge, and half lives were 23.9 days and 36.5 days respectively. The optimal pH for NP degradation in sludge was 7 and the degradation rate was enhanced when the temperature was increased. The addition of yeast extract (5 mg/l) or surfactants such as brij 30 or brij 35 (55 or 91 microM) also enhanced the NP degradation rate. The addition of aluminum sulfate (200 mg/l) inhibited the NP degradation rate within 84 days of incubation. The high-to-low order of degradation rates was: sulfate-reducing conditions>methanogenic conditions>nitrate-reducing conditions. Sulfate-reducing bacteria, methanogen, and eubacteria are involved in the degradation of NP, sulfate-reducing bacteria being a major component of sludge. PMID- 15876385 TI - Prediction of octanol--air partition coefficients of semivolatile organic compounds based on molecular connectivity index. AB - A new method has been developed to describe the quantitative relationship between the octanol-air partition coefficients and molecular connectivity indexes of semivolatile organic compounds such as polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated naphthalenes (PCNs), polycyclic aromatic hydrocarbons (PAHs), chlorobenzenes (CBs), polybrominated diphenyl ethers (PBDEs). The stepwise multiple linear regression was used to derive six equations with correlation coefficients greater than 0.99 except for PBDEs' (0.96). The largest relative error is 4.6%, and the standard deviation is less than 1.83 log units. Compared with the data obtained from the relevant experiments, the results of prediction are very accurate. PMID- 15876386 TI - Removal of sorbed polycyclic aromatic hydrocarbons from soil, sludge and sediment samples using the Fenton's reagent process. AB - The use of the Fenton's reagent process has been investigated for the remediation of environmental matrices contaminated by polycyclic aromatic hydrocarbons (PAHs). Laboratory experiments were first conducted in aqueous solutions, to study the kinetics of oxidation and adsorption of PAHs. Benzo[a]pyrene was more rapidly degraded than adsorbed, while only partial oxidation of fluoranthene occurred. In the case of benzo[b]fluoranthene, its adsorption prevented its oxidation. Besides competition effects between PAHs were found, with slower oxidation of mixtures as compared to single PAH solutions. Apparition of some by products was observed, and a di-hydroxylated derivative of benzo[a]pyrene could be identified under our conditions. Consequently, application to solid environmental matrices (soil, sludge and sediment samples) was performed using large amounts of reagents. The efficiency of the Fenton treatment was dependent on the matrix characteristics (such as its organic carbon content) and the PAH availability (correlated to the date and level of contamination). However, no pH adjustment was required, as well as no iron addition due to the presence of iron oxides in the solid matrices, suggesting the potential application of Fenton-like treatment for the remediation of PAH-contaminated environmental solids. PMID- 15876387 TI - Gas-particle partitioning of PCDD/Fs in Nagoya urban air, Japan. AB - Polychlorinated dibenzo-p-dioxins and -furans (PCDD/Fs) in the atmosphere were sampled by a high-volume filter/sorbent air sampler at an urban (Nagoya, Japan) site from May 2000 to January 2001 under the average ambient temperatures ranging from 1 degrees C to 33 degrees C. The gas and particle phase concentrations of PCDD/Fs were measured and then the field gas-particle partitioning data were used to assess the Junge-Pankow adsorption model and the K(OA) absorption model. For both PCDDs and PCDFs, passable agreement was obtained between the measured and predicted particulate-band fraction (phi) values by the Junge-Pankow model, but the model tends to overpredict phi for all individual Cl4-8DD/F congeners, especially for the lower chlorinated congeners. On the other hand, good agreement was obtained between the measured and predicted log K(p) values by the K(OA) model for both PCDDs and PCDFs. However, the K(OA) absorption model did not fit the field data at low ambient temperature below 5 degrees C. The disagreement at low ambient temperatures was estimated to arise from the gas adsorption artifacts during the sampling. From the validation by the field measurement data in Nagoya urban air, it is found that the K(OA) absorption model is capable of predicting more accurately gas-particle partitioning of PCDD/Fs than the Junge-Pankow adsorption model. PMID- 15876388 TI - Matrix-immobilized organoclay for the sorption of polycyclic aromatic hydrocarbons and pentachlorophenol from groundwater. AB - Sorbent materials consisting of organoclay immobilized onto the surface of a solid support were evaluated for use in pentachlorophenol (PCP) and polycyclic aromatic hydrocarbon (PAH) remediation of groundwater at a creosote-contaminated Superfund site. Cetylpyridinium-exchanged low pH montmorillonite clay (CP-LPHM) was bonded to either sand (CP-LPHM/sand) or granular activated carbon (GAC) (CP LPHM/GAC) using the free acid form of carboxymethylcellulose as an adhesive. Effluent from an oil-water separator was eluted through equal bed volumes of composite (4 g 3:2 CP-LPHM/GAC or 13 g CP-LPHM/sand), affinity-extracted, and quantitatively analyzed by GC/MS. PCP, naphthalene, fluorene, phenanthrene, pyrene, and total PAHs were initially reduced by both CP-LPHM/GAC (> or =99%, 61%, 99%, > or =99%, 97%, and 94%, respectively) and CP-LPHM/sand (90%, 70%, 94%, 95%, 93%, and 86%, respectively). Complete breakthrough of naphthalene occurred after approximately 15 h of elution through 3:2 CP-LPHM/GAC and 22 h through CP LPHM/sand. PCP showed complete breakthrough following 18 h of elution through 3:2 CP-LPHM/GAC and 26 h through CP-LPHM/sand. However, 50% breakthrough was not attained for higher molecular weight PAHs, as fluoranthene, pyrene, benzo[a]anthracene, and chrysene continued to be greatly reduced with both 3:2 CP LPHM/GAC (98%, 95%, 94%, and 95%, respectively) and CP-LPHM/sand (75%, 73%, 76%, and 78%, respectively) after 48 h of continuous elution. Results confirm prior studies, indicating that these organoclay-containing composites have a high capacity for contaminants found in wood preserving waste. Further, results suggest that the inclusion of CP-LPHM may be useful as part of an effective strategy for groundwater remediation of high concentrations of PCP and PAHs, in particular high molecular weight and carcinogenic PAHs. PMID- 15876389 TI - Evaluation of PCDD/Fs patterns emitted from incinerator via direct ambient sampling and indirect serum levels assessment of Taiwanese. AB - The aim of this study was to evaluate the PCDD/Fs patterns in ambient air based on data information emitted from incinerator generated from ambient air measurements and those in serum. Four circular zones, namely A, B, C, and D, were identified based on simulated ambient annual average PCDD/Fs concentrations, from a selected municipal waste incinerator. Sixteen ambient samples were taken from the 4 circular zones across 4-seasons. Eighty-nine volunteers were recruited according to the demographic distribution within each zone. PCDD/Fs profiles were documented both for air and serum samples collected. Comparing to the congener patterns from ambient air and serum samples, we found that OCDD, OCDF, 1,2,3,4,6,7,8-HpCDD, and 1,2,3,4,6,7,8-HpCDF were the predominant groups among 17 congeners from both the ambient air and serum sample. And, factor analysis showed the distribution patterns of PCDD/Fs from ambient air and serum samples are almost identical across different zones, except for congener patterns of serum samples from residents in zone C. In addition, the average PCDD/Fs level significantly reduced for about 10 folds than those of the other three seasons when the incinerator was shut down in one of sampling periods. We might conclude that ambient air exposure was the most important contributor to PCDD/Fs levels in ambient air but not the single in serum. Therefore, another or more powerful source, such as occupational exposure, dietary intake or the consumption of local food, should be further investigated at the same time. PMID- 15876390 TI - Testing common sediment-porewater distribution models for their ability to predict dissolved concentrations of POPs in The Grenlandsfjords, Norway. AB - This study compares in situ observed porewater concentration of persistent organic pollutants (POPs) with predictions by common solid-water phase distribution models. Bottom sediments were sampled in The Grenlandsfjords, Norway, and the interstitial porewater was isolated from the solids by centrifugation and filtration. Both phases were analysed for polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/F), polycyclic aromatic hydrocarbons (PAH), and organic carbon. Based on the sediments' solid phase content of POPs, organic carbon, and soot carbon, we used the organic matter partitioning (OMP), and also the soot and organic matter partitioning (SOMP) model to estimate the porewater concentration. The OMP model gave better agreement to observations than the SOMP model for both PCDD/Fs and PAHs. The observed concentration of the PCDD/Fs in the sediments' porewater was much higher than in the deep water of the fjord. The logarithm of the organic matter-water partitioning coefficent (log K(OC)) in the porewater had positive linear regression on the logarithm of the octanol-water partitioning coefficient (log K(OW)). The slope of the regression model was indistinguishable from 1, except for the PAHs as a group which had a slope less than 1. This contrasts to previous studies undertaken in The Grenlandsfjords water column, where the slopes were higher than 1 for PCDD/Fs, and the K(OC) were much higher than the K(OW). One explanation may be that the influence of POPs adsorption to soots decrease because competitive sorption by other compounds in the sediment are higher than in the water column. This indicates that the sorption isotherms for these POPs need better understanding in order to be applicable in both the water column and the porewater. PMID- 15876391 TI - Dechlorination of polychlorinated organic compounds by electrochemical reduction with naphthalene radical anion as mediator. AB - Electrochemical reduction with electrochemically generated naphthalene radical anion in N,N-dimethylformamide was applied to the dechlorination of five representative POPs, namely HCB, lindane, DDT, PCP and aldrin. Rapid and complete dechlorination was possible for lindane and DDT to give nearly quantitative yields of benzene and 1,1-diphenylethane, respectively. HCB was reduced through complex reaction pathways to yield unknown products. Dechlorination of PCP and aldrin beyond dichlorinated compounds was difficult because of their very negative reduction potential. The reaction pathways for each dechlorination were proposed with the identification of intermediates. PMID- 15876392 TI - The effect of copper speciation on the formation of chlorinated aromatics on real municipal solid waste incinerator fly ash. AB - A limited amount of information exists regarding the relationship between the chemical form of copper and the formation of chlorinated aromatics in fly ash. To understand the effects of the various forms of copper on the formation of chlorinated aromatics in real fly ash, we determined the chemical forms of copper present in various types of real fly ash using X-ray absorption near edge structure (XANES) and evaluated the relationship between the chemical forms of copper and the formation of chlorinated aromatics. Copper chloride hydroxide (CuCl2 x 3Cu(OH)2) and cuprous chloride (CuCl) were the predominant copper species found in real fly ash. Although pure cupric chloride (CuCl2) is known to be the most active catalyst for the formation of chlorinated aromatics under experimental conditions with synthetic fly ash, CuCl2 was not found in every real fly ash sample. The amount of copper chloride hydroxide was positively correlated with the formation of chlorinated aromatics in real fly ash and is, consequently, considered to be one of the key species involved in the formation of chlorinated aromatics. PMID- 15876393 TI - Persistent organic pollutants in soil density fractions: distribution and sorption strength. AB - The sorption strength of persistent organic pollutants in soils may vary among different soil organic matter (SOM) pools. We hypothesized that polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were unevenly distributed and had different soil organic carbon (SOC)-water partition coefficients (K(OC)) among soil density fractions. We determined the concentrations and K(OC) values of 20 PAHs and 12 PCBs in bulk samples and three density fractions (light, <2.0, medium, 2.0-2.4, and heavy, >2.4 g cm(-3)) of 11 urban topsoils (0-5 cm) from Bayreuth, Germany. The K(OC) values were determined using sequential extraction with methanol-water mixtures (35% and 65% methanol) at 60 degrees C. The sum of 20 PAH concentrations in bulk soil ranged 0.4-186 mg kg(-1), and that of 12 PCB concentrations 1.2-158 microg kg(-1). The concentrations of all PAHs and PCBs decreased in the order light>medium>heavy fraction. When normalized to the SOC concentrations, PAH concentrations were significantly higher in the heavy than in the other density fractions. The K(OC) values of the PAHs in density fractions were 3-20 times higher than those of the PCBs with similar octanol-water partition coefficients (K(OW)). The K(OC) values of individual PAHs and PCBs varied up to a factor of 1000 among the studied soils and density fractions. The K(OC) values of 5- and 6-ring PAHs tended to be highest in the heavy fraction, coinciding with their enrichment in this fraction. For the other PAHs and all PCBs, the K(OC) values did not differ among the density fractions. Thus, there is no relationship between sorption strength and distribution among density fractions, indicating that density fractionation is not a suitable tool to distinguish among differently reactive PAH and PCB pools in soils. PMID- 15876394 TI - Occurrence of PCDD/Fs in urban air before and after the ban of leaded gasoline. AB - The source of PCDDs and PCDFs in automotive exhaust is not yet fully explained. The chlorinated hydrocarbons used in the formulation of lead-alkyl additives were suspected as a possible major source. Based on this, the decreasing use of leaded gasoline followed by its final ban (occurred on 1/1/2002, in Italy) should have resulted in a decreasing presence of PCDD/Fs in urban air and possibly some differences in their profile. To investigate these aspects, we monitored PCDD/Fs for one year starting in September 2001, at a medium-traffic road site in Rome, with weekly frequency. Results were then compared with those obtained in a previous study performed before the ban (from February 2000 to January 2001) at the same site. As compared with the previous study, the yearly-averaged overall PCDD/F concentration, as toxic equivalent of 2,3,7,8-TCDD, decreased from 60 to 50 fg m(-3) I-TEQ; it remained unchanged, however, if one sample with a particularly high PCDD/F content was excluded from each data set. The monthly trend confirmed the one found in the former study. On an annual basis, the two mean congener profiles were almost identical. The concentration levels and the constancy of profiles, as calculated for the two periods, do not support the hypothesis of a major role of leaded gasoline, substantially different from unleaded one, in contributing to PCDD/F air pollution. PMID- 15876395 TI - Targeted drug delivery for musculoskeletal diseases. PMID- 15876396 TI - Quest for the Holy Grail to cure arthritis and osteoporosis: emphasis on bone drug delivery systems. AB - The number of new medications to treat and even prevent arthritis and osteoporosis has expanded dramatically in recent years. Where once there were only surgical options to treat such end-stage diseases, there are now treatments targeted at the early steps in musculoskeletal pathophysiology. The use of different modalities to maximize drug access to specific bone tissues has created a golden opportunity for mechanistic studies in drug delivery systems for treating osteoporosis and other musculoskeletal diseases. This theme issue provides a timely analysis of the challenges and accomplishments in delivering medicine to the target sites in the musculoskeletal system and also provides a preview of what may come in the future for musculoskeletal medicine. As the number of animal studies and clinical trials is on the rise, the possibility to prevent or even cure the aforementioned disorders has never been closer. PMID- 15876397 TI - Skeletal function and structure: implications for tissue-targeted therapeutics. AB - Osteoporosis, arthritis, and periodontal disease are common diseases of the skeleton, all of which could benefit from new therapeutic strategies, including targeted drug delivery. While bone is a rigid structure, it is not inert, with the cells of the skeleton being able to repair damage and respond to alterations in mechanical stimuli and various endocrine agents. Several important factors related to bone physiology that could influence the success of a pharmacological treatment include heterogeneity in bone remodeling activities throughout the skeleton, differences in blood supply and local vascularization, and the "blood bone" barrier. The structural qualities of bone, especially the presence of hydroxyapatite crystals in the bone mineral and the established binding of certain molecules to this mineral phase, including tetracyclines, bisphosphonates, and other chelators, provide unique opportunities to treat skeletal diseases using targeted drug delivery. Additional opportunities exist in targeting sites with contrasting bone surface activities, including surfaces that are inactive, forming new bone or being resorbed. The ultimate key to developing new bone-targeted therapies is to understand and exploit the physiological characteristics at the desired target sites. PMID- 15876398 TI - Mechanism of osteoclast mediated bone resorption--rationale for the design of new therapeutics. AB - Bone resorption is an important cellular function in the development and physiology of the skeleton. Pathophysiology of several skeletal diseases includes either increased (for instance osteoporosis, metastatic bone disease and Paget's disease of bone) or decreased (various syndromes of osteopetrosis) bone resorption rate. Thus there is a genuine need to regulate, especially inhibit, bone resorption rate in several diseases. Bone resorption can be inhibited by several strategies. One can prevent osteoclast formation, inhibit their action or induce premature cell death. All these strategies have been used in pharmacology to inhibit bone resorption and there are also physiological regulators for each of these three different phases of in osteoclast life. Many present resorption inhibitors inhibit osteoclast formation via osteoblastic cells since they are producing a number of factors that are essential for osteoclast differentiation. Best characterized of these factors are macrophage colony stimulating factor (M CSF) and receptor activator of NFeB ligand (RANKL). For instance sex steroids, parathyroid hormone and some interleukins are known to exert their positive or negative effects on osteoclast differentiation via the RANK/RANKL/osteoprotegrin pathway. It is not yet clear enough how specific intervention to osteoclast formation is since also other cell lineages derived from hematopoetic precursors use similar signalling pathways. An effective inhibition of bone resorption can also be achieved by inhibiting osteoclast activity to resorb bone. Examples of this category of physiological and pharmacological inhibitors are calcitonin and aminobisphosphonates, respectively. Finally one can reduce bone resorption by shortening osteoclast lifespan with substances that induce apoptosis in osteoclasts. A good example of these substances is a first generation bisphosphonate, clodronate. Several new potential molecular targets have been revealed during recent years since many individual molecules in osteoclast differentiation, function and apoptosis have been identified and their physiological functions revealed. Thus we are expecting several new bone inhibitors to be developed in following years both for experimental studies and finally also for clinical use. PMID- 15876399 TI - The role of cathepsins in osteoporosis and arthritis: rationale for the design of new therapeutics. AB - Human cysteine proteases of the papain family have been recognized as potential drug targets for musculoskeletal diseases. Most of the interest is focused on cathepsins S and K, which display selective expression in cells of the immune system and cells capable to efficiently degrade extracellular matrix proteins, in particular collagens. The predominant expression of cathepsin K in osteoclasts has rendered the enzyme into a major target for the development of novel anti resorptive drugs in osteoporosis whereas cathepsin S appears to be an attractive drug target candidate for various inflammatory diseases including rheumatoid arthritis. Since rheumatoid arthritis is at the same time an inflammatory and joint destructive disorder, the combined inhibition of both cathepsins S and K should be beneficial. This review will outline the rationale and recent progress for targeting cathepsins in arthritis and osteoporosis. PMID- 15876400 TI - Targeting of therapeutic agents to bone to treat metastatic cancer. AB - The three main organs affected by metastasis of all cancers include lungs, liver, and bone. Clinical confirmation of tumor spread to these organs is a negative prognostic sign that marks the stage when disease is rarely curable. Today, treatment of bone metastases is primarily palliative. The aims of treatment are to relieve pain, prevent development of pathologic fractures, improve mobility and function, and if possible, prolong survival. Significant improvements in our understanding of tumor biology along with early tumor detection has led to the discovery of few innovative approaches aimed to treat bone metastases. The most promising treatment modalities include combination of anti-cancer therapies (surgery, radiation therapy, citostatic therapy) with bone antiresorptive therapies (bisphosphonate) that specifically target osteoclasts, bone resorbing cells. The osteoclast, whose increased activity is induced by the tumor, is responsible for the deterioration of bone mass and structure along with the release of grow factors that feed back and support further tumor growth. The current pharmaceutical approach is to target bone metastases by developing drugs that specifically target tumor cells in bone in addition to bone stroma since skeletal metastases are more resistant to treatment, present the highest bulk of tumor mass in the body, serve as site for secondary spread of tumor cells, and are associated with significant morbidity. There is a real need for a more effective modified release of newer anti-cancer drugs such as gene therapy and immunotherapy by using established and novel delivery platforms that will improve therapy and reduce side effects as a result of more appropriate plasma profiles. Overall, however, developments regarding treatment of cancer metastases to bone are encouraging. The scope of future advancements is immense and includes innovative therapeutics and delivery systems aimed to improve skeletal affinity, selectivity, and efficacy of drugs. PMID- 15876401 TI - Designing proteins for bone targeting. AB - Protein-based therapeutic agents intended for bone diseases should ideally exhibit a high affinity to bone tissue, so that their systemic administration will result in specific delivery to bone with minimal distribution to extra skeletal sites. This was shown possible in the authors' lab by modifying a desired protein with bisphosphonates (BPs) that exhibit an exceptionally high affinity to the bone-mineral hydroxyapatite. In this review, we explore the potential applications of that concept by summarizing the bone diseases and candidate proteins that will benefit from the proposed bone delivery approach. A selective synopsis of BP synthesis is presented to highlight the synthesis of functional BPs suitable for covalent attachment to proteins. Finally, we present a summary of recent research results from the authors' laboratory emphasizing factors influencing bone affinity of the conjugates. We conclude with future research avenues that are considered critical for clinical entry of the BP targeted therapeutic agents. PMID- 15876402 TI - Synthetic biodegradable polymers as drug delivery systems for bone morphogenetic proteins. AB - Bone morphogenetic proteins (BMP) induce bone formation in vivo, and clinical application in repair of bone fractures and defects is expected. However, appropriate systems to deliver BMP for clinical use need to be developed. We synthesized a new synthetic biodegradable polymer, poly-D,L-lactic acid-para dioxanone-polyethylene glycol block copolymer (PLA-DX-PEG), to serve as a biocompatible, biodegradable polymer for recombinant human (rh) BMP-2 delivery systems. In animal experiments, new bone was efficiently formed and a large bone defect was repaired using PLA-DX-PEG/rhBMP-2 composites. In addition, this new polymer could be used as an injectable delivery system for rhBMP-2. The rhBMP 2/PLA-DX-PEG composites also could be combined with other materials such as hydroxyapatite or titanium. This new synthetic polymer might be used for rhBMP-2 delivery in various clinical situations involving repair of bone, leading to great changes in orthopedic treatment. PMID- 15876403 TI - Bone-targeting macromolecular therapeutics. AB - Musculoskeletal diseases such as osteoporosis are recognized as major public health problems worldwide. Many novel therapeutic agents have been identified for the treatment of these diseases. However, the majority of them are not specific to hard tissue, resulting significant toxicity. Bone-targeting drug delivery systems based on water-soluble polymers can specifically direct candidate drugs to bone thereby reducing side effects due to non-specific tissue interactions. Incorporation of a targeting moiety, a drug release mechanism, drug selection and optimization of the polymer carrier are all essential elements in the development of bone-targeting macromolecular therapeutics. Successful clinical application of this approach can significantly contribute to the development of treatments for many musculoskeletal diseases. PMID- 15876404 TI - Gene regulation by the glucocorticoid receptor: structure:function relationship. AB - The glucocorticoid receptor (GR) belongs to the superfamily of ligand-activated transcription factors, the nuclear hormone receptors. Like other members of the family, the GR possesses a modular structure consisting of three major domains the N-terminal (NTD), DNA binding (DBD), and ligand binding (LBD). Although the structures of independently expressed GR DBD and LBD are known, the structures of the NTD and of full-length GR are lacking. Both DBD and LBD possess overall globular structures. Not much is known about the structure of the NTD, which contains the powerful AF1/tau1/enh2 transactivation region. Several studies have shown that AF1 region is mostly unstructured and that it can acquire folded functional conformation under certain potentially physiological conditions, namely in the presence of osmolytes, when the GR DBD is bound to glucocorticoid response element (GRE), and when AF1 binds other transcription factor proteins. These conditions are discussed here. The functions of the GR will be fully understood only when its working three-dimensional structure is known. Based on the available data, we propose a model to explain data which are not adequately accounted for in the classical models of GR action. In this review, we summarize and discuss current information on the structure of the GR in the context of its functional aspects, such as protein:DNA and protein:protein interactions. Because of the close similarities in modular organization among the members of the nuclear hormone receptors, the principles discussed here for the GR should be applicable to many other receptors in the family as well. PMID- 15876405 TI - Sex change strategy and the aromatase genes. AB - Sequential hermaphroditism is a common reproductive strategy in many teleosts. Steroid production is known to mediate both the natural and induced sex change, yet beyond this the physiology directing this process has received little attention. Cytochrome P450 aromatase is a key enzyme in the hormonal pathway catalysing the conversion of sex steroids, androgens to oestrogens, and thus is highly relevant to the process of sex change. This study reports the isolation of cDNA sequences for aromatase isoforms CYP19A1 and CYP19A2 from teleost species representing three forms of sexual hermaphroditism: Lates calcarifer (protandry), Cromileptes altivelis (protogyny), and Gobiodon histrio (bi-directional). Deduced amino acid analysis of these isoforms with other reported isoforms from gonochoristic (single sex) teleosts revealed 56-95% identity within the same isoform while only 48-65% identity between isoforms irrespective of species and sexual strategy. Phylogenetic analysis supported this result separating sequences into isoform exclusive clades in spite of species apparent evolutionary distance. Furthermore, this study isolates 5' flanking regions of all above genes and describes putative cis-acting elements therein. Elements identified include steroidogenic factor 1 binding site (SF-1), oestrogen response element (ERE), progesterone response element (PRE), androgen response element (ARE), glucocorticoid response elements (GRE), peroxisome proliferator-activated receptor alpha/retinoid X receptor alpha heterodimer responsive element (PPARalpha/RXRalpha), nuclear factor kappabeta (NF-kappabeta), SOX 5, SOX 9, and Wilms tumor suppressor (WTI). A hypothetical in vivo model was constructed for both isoforms highlighting potential roles of these putative cis-acting elements with reference to normal function and sexual hermaphroditism. PMID- 15876406 TI - New assumptions about oxidative processes involved in steroid hormone biosynthesis: is the role of cytochrome P-450-activated dioxygen limited to hydroxylation reactions or are dioxygen insertion reactions also possible? AB - The traditional conception of the chemical pathways leading to the formation of the steroid hormones is derived by piecing together the results of several independent in vitro incubation experiments. The results of these experiments have led to the assumption that some relevant cytochrome P-450's (P-450scc, P 450arom, P-450aldo, etc.) are "polyfunctional" and catalyze several successive hydroxylation reactions, which lead to the formation of the hormonal products. This essay offers an alternative view. It advances the suggestion that the oxygenated intermediates in the relevant biosynthetic conversions are reactive species that are formed by addition of both atoms of dioxygen onto two neighboring carbon atoms of steroidal precursors. Space-filled Stuart molecular models, generated by a computer program, suggest that the oxidized intermediates resemble hydroperoxides or cyclic peroxides (1,2-dioxanes). For the aromatization process required for estrogen biosynthesis, the atoms of dioxygen are bonded to C 2 and C-19 of the C19-precursor. For aldosterone formation, dioxygen is bonded to C-11 and C-18 of an appropriate precursor. Moreover, the results obtained from a computer program that provides information about "molecular mechanics" (bond angles and bond distances as well as total potential energies for each conformation of a molecule) suggest that consideration be given to the possibility that cortisol also can be biosynthesized by P-450-activated dioxygen addition to C-11 and C-17 of an appropriate precursor. Neither the traditional view of steroidogenic pathways nor the suggestions advanced here have been established by compelling experimental findings. Both hypotheses are saddled with untested assumptions, which are necessary because the dynamic processes can only be discerned by indirect means. The origins of some naturally occurring steroids hydroxylated at C-17, C-18 and C-19 are examined in the light of the suggestions made in this essay. PMID- 15876407 TI - Attainment of peak bone mass and bone turnover rate in relation to estrous cycle, pregnancy and lactation in colony-bred Sprague-Dawley rats: suitability for studies on pathophysiology of bone and therapeutic measures for its management. AB - Alteration in biochemical markers of bone turnover and bone mineral density (BMD) of whole body and isolated femur and tibia in relation to age, estrous cycle, pregnancy and lactation and suitability of use of rat as model for studies on pathophysiology of bone and therapeutic measures for its management were investigated. Immature rats (1, 1.5 and 2 month of age; weighing, respectively, 39.3+/-1.0, 67.8+/-2.4 and 87.2+/-5.2 g) exhibited high rate of bone turnover, as evidenced by high serum osteocalcin and alkaline phosphatase and urine calcium/creatinine ratio. However, their BMD (whole body or of isolated long bones) was below measurable levels. Marked increase in body weight at 3 months (185.5+/-5.2 g) was associated with low serum osteocalcin and alkaline phosphatase and urine calcium/creatinine ratio. Biochemical markers and BMD attained at puberty at 3 months were maintained until 36 month of age. No significant change in serum calcium was observed with increasing age or on any of the biomarkers during estrous cycle, and BMD of femur and tibia isolated during proestrus and diestrus stages was almost similar. Onset of pregnancy was associated with significant increase in serum total alkaline phosphatase and osteocalcin levels, but serum calcium, urine calcium/creatinine ratio or BMD of whole body or isolated long bones were not significantly different from that at proestrus stage. No marked change, except increase in body weight (P<0.05), was also evident in these parameters between days 5 and 19 of pregnancy, irrespective of number of implantations in the uterus. A significant decrease in BMD of isolated femur (neck and mid-shaft regions) was observed on days 5 and 21 of lactation as compared to that during pregnancy or diestrus/proestrus stages of estrous cycle; the decrease being almost similar in females lactating two or six young ones. BMD of isolated tibia (global and region proximal to tibio-fibular separation point), though generally lower than that during cycle and pregnancy, was statistically non-significant. However, clear evidence of occurrence of osteoporosis during lactation, with decrease in BMD of >2.5 x S.D. in isolated femur (global, neck and mid-shaft) as well as tibia (global) was observed only when BMD data was analysed on T-/Z-score basis. Serum biochemical markers of bone turnover, too, were significantly increased in comparison to cyclic rats. Findings demonstrate marked increase in body weight and bone turnover during first 3 months of age, direct correlation between peak bone mass and onset of puberty at 3 months of age and increase in bone resorption rate during lactation. Finding of the study while might suggests possible use of rat as useful model for studies on bone turnover rate during lactation and post-weaning periods and extrapolation of the result to the human situation, but not in relation to ageing. PMID- 15876408 TI - Comparative study of oestrogenic properties of eight phytoestrogens in MCF7 human breast cancer cells. AB - Previous studies have compared the oestrogenic properties of phytoestrogens in a wide variety of disparate assays. Since not all phytoestrogens have been tested in each assay, this makes inter-study comparisons and ranking oestrogenic potency difficult. In this report, we have compared the oestrogen agonist and antagonist activity of eight phytoestrogens (genistein, daidzein, equol, miroestrol, deoxymiroestrol, 8-prenylnaringenin, coumestrol and resveratrol) in a range of assays all based within the same receptor and cellular context of the MCF7 human breast cancer cell line. The relative binding of each phytoestrogen to oestrogen receptor (ER) of MCF7 cytosol was calculated from the molar excess needed for 50% inhibition of 3H]oestradiol binding (IC50), and was in the order coumestrol (35x)/8-prenylnaringenin (45x)/deoxymiroestrol (50x)>miroestrol (260x)>genistein (1000x)>equol (4000x)>daidzein (not achieved: 40% inhibition at 10(4)-fold molar excess)>resveratrol (not achieved: 10% inhibition at 10(5)-fold molar excess). For cell-based assays, the rank order of potency (estimated in terms of the concentration needed to achieve a response equivalent to 50% of that found with 17beta-oestradiol (IC50)) remained very similar for all the assays whether measuring ligand ability to induce a stably transfected oestrogen-responsive ERE CAT reporter gene, cell growth in terms of proliferation rate after 7 days or cell growth in terms of saturation density after 14 days. The IC50 values for these three assays in order were for 17beta-oestradiol (1 x 10(-11)M, 1 x 10( 11)M, 2 x 10(-11)M), and in rank order of potency for the phytoestrogens, deoxymiroestrol (1 x 10(-10)M, 3 x 10(-11)M, 2 x 10(-11)M)>miroestrol (3 x 10( 10)M, 2 x 10(-10)M, 8 x 10(-11)M)>8-prenylnaringenin (1 x 10(-9)M, 3 x 10(-10)M, 3 x 10(-10)M)>coumestrol (3 x 10(-8)M, 2 x 10(-8)M, 3 x 10(-8)M)>genistein (4 x 10(-8)M, 2 x 10(-8)M, 1 x 10(-8)M)/equol (1 x 10(-7)M, 3 x 10(-8)M, 2 x 10( 8)M)>daidzein (3 x 10(-7)M, 2 x 10(-7)M, 4 x 10(-8)M)>resveratrol (4 x 10(-6)M, not achieved, not achieved). Despite using the same receptor context of the MCF7 cells, this rank order differed from that determined from receptor binding. The most marked difference was for coumestrol and 8-prenylnaringenin which both displayed a relatively potent ability to displace [3H]oestradiol from cytosolic ER compared with their much lower activity in the cell-based assays. Albeit at varying concentrations, seven of the eight phytoestrogens (all except resveratrol) gave similar maximal responses to that given by 17beta-oestradiol in cell-based assays which makes them full oestrogen agonists. We found no evidence for any oestrogen antagonist action of any of these phytoestrogens at concentrations of up to 10(-6)M on either reporter gene induction or on stimulation of cell growth. PMID- 15876409 TI - Combinatory effects of phytoestrogens and 17beta-estradiol on proliferation and apoptosis in MCF-7 breast cancer cells. AB - Phytoestrogens have been described to be weak estrogens, SERMs or exhibit antiestrogenic properties. However, information about their activity in presence of estrogens is limited. Therefore, we have analysed the dose dependent combinatory activity of the phytoestrogens genistein (Gen), daidzein (Dai) and coumestrol (Cou), and 17beta-estradiol (E2) on cell proliferation and apoptosis induction in human MCF-7 breast cancer cells. Neither additive nor antagonistic effects on proliferation could be observed, but in contrast all phytoestrogens possessed the ability to inhibit apoptosis in the presence of 17beta-estradiol. In summary, our in vitro results demonstrate that Gen does not exhibit any antiestrogenic properties. The additive growth stimulatory effects of Gen, Dai and Cou in the presence of E2 are not the result of a stimulation of proliferation; these phytoestrogens, at least in MCF-7 cells, could be characterised as inhibitors of apoptosis. PMID- 15876410 TI - Anti-growth factor activities of benzothiophenes in human breast cancer cells. AB - We have tested the effects of two Eli-Lilly compounds, LY 117, 018 and raloxifene, on E2-regulated and IGF-I-induced proliferation or AP-1 activity in human breast cancer cells. We now demonstrate that both molecules have strong antiestrogenic and anti-growth factor inhibitory effects in MCF7 cells. They were as potent as ICI 182, 780 and more efficient than OH-Tam to prevent estradiol action whereas their inhibition on IGF-I stimulation was less than with ICI 182, 780 and equivalent to that of OH-Tam. Moreover, raloxifene was the most efficient molecule to prevent IGF-I-induced AP-1 activity, with a significant effect observed with a concentration as low as 5 x 10(-11)M in the presence of IGF-I alone. Similar dose-response curves were obtained with a combined treatment of IGF-I and E2 with a 2log shift. Their action on IGF-I-induced proliferation was completely abrogated in MCF7 transfectants in which the expression of an antiestrogen-regulated protein tyrosine phosphatase, PTPL1, was abolished by antisense RNA transfection. Accordingly, they were both able to dose-dependently regulate the expression of PTPL1 and to interfere with the PI3-K/Akt pathway by drastically decreasing Akt phosphorylation exclusively in wild-type PTPL1 expressing cells. Our data altogether demonstrate that raloxifene has a potent inhibitory effect on IGF-I action, with a drastic effect on AP-1 triggered responses as well as on Akt phosphorylation, suggesting that it might be a useful therapeutic agent in tumors in which these signalling pathways become constitutively active. PMID- 15876411 TI - Mammalian lignans and genistein decrease the activities of aromatase and 17beta hydroxysteroid dehydrogenase in MCF-7 cells. AB - Estrogen plays a major role in breast cancer development and progression. Breast tissue and cell lines contain the necessary enzymes for estrogen synthesis, including aromatase and 17beta-hydroxysteroid dehydrogenase (17beta-HSD). These enzymes can influence tissue exposure to estrogen and therefore have become targets for breast cancer treatment and prevention. This study determined whether the isoflavone genistein (GEN) and the mammalian lignans enterolactone (EL) and enterodiol (ED) would inhibit the activity of aromatase and 17beta-HSD type 1 in MCF-7 cancer cells, thereby decreasing the amount of estradiol (E2) produced and consequently cell proliferation. Results showed that 10 microM EL, ED and GEN significantly decreased the amount of estrone (E1) produced via the aromatase pathway by 37%, 81% and 70%, respectively. Regarding 17beta-HSD type 1, 50 microM EL and GEN maximally inhibited E2 production by 84% and 59%, respectively. The reduction in E1 and E2 production by EL and the reduction in E2 production by GEN were significantly related to a reduction in MCF-7 cell proliferation. 4 Hydroxyandrostene-3,17-dione (50 microM) did not inhibit aromatase but inhibited the conversion of E1 to E2 by 78%, suggesting that it is a 17beta-HSD type 1 inhibitor. In conclusion, modulation of local E2 synthesis is one potential mechanism through which ED, EL and GEN may protect against breast cancer. PMID- 15876412 TI - The antagonism between 2-methyl-1,25-dihydroxyvitamin D3 and 2-methyl-20-epi-1,25 dihydroxyvitamin D3 in non-genomic pathway-mediated biological responses induced by 1alpha,25-dihydroxyvitamin D3 assessed by NB4 cell differentiation. AB - We synthesized all eight possible A-ring diastereomers of 2-methyl substituted analogs of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] and also all eight A ring diastereomers of 2-methyl-20-epi-1alpha,25(OH)2D3. Their biological activities, especially the antagonistic effect on non-genomic pathway-mediated responses induced by 1alpha,25(OH)2D3 or its 6-s-cis-conformer analog, 1alpha,25(OH)2-lumisterol3, were assessed using an NB4 cell differentiation system. Antagonistic activity was observed for the 1beta-hydroxyl diastereomers, including 2beta-methyl-1beta,25(OH)2D3 and 2beta-methyl-3-epi-1beta,25(OH)2D3. Very interestingly, 2beta-methyl-3-epi-1alpha,25(OH)2D3 also antagonized the non genomic pathway, despite its 1alpha-hydroxyl group. Other 1alpha-hydroxyl diastereomers did not show antagonistic activity. 20-epimerization diminished the antagonistic effect of all of these analogs on the non-genomic pathway. These findings suggested that the combination of the 2-methyl substitution of the A ring and 20-epimerization of the side chain could alter the biological activities in terms of antagonism of non-genomic pathway-mediated biological response. Based on a previous report, 2-methyl substitution alters the equilibrium of the A-ring conformation between the alpha- and beta-chair conformers. The 2beta-methyl diastereomers, which exhibited antagonism on non-genomic pathway-mediated response, were considered to prefer the beta-conformer. Further examination to elucidate the relationship between the altered ligand shape and receptors interaction will be important for molecular level understanding of the mechanism of antagonism of the non-genomic pathway. PMID- 15876413 TI - Anabolic-androgenic steroid interaction with rat androgen receptor in vivo and in vitro: a comparative study. AB - Anabolic steroids are synthetic derivatives of testosterone and are characterized by their ability to cause nitrogen retention and positive protein metabolism, thereby leading to increased protein synthesis and muscle mass. There are disagreements in the literature in regards to the interaction of anabolic steroids with the androgen receptor (AR) as revealed by competitive ligand binding assays in vitro using cytosolic preparations from prostate and skeletal muscle. By use of tissue extracts, it has been shown that some anabolic steroids have binding affinities for the AR that are higher than that of the natural androgen testosterone, while others such as stanozolol and methanedienone have significantly lower affinities as compared with testosterone. In this study we show that stanozolol and methanedienone are low affinity ligands of the rat recombinant AR as revealed by a ligand binding assay in vitro, however, based on a cell-based AR-dependent transactivation assay, they are potent activators of the AR. We also show that a single injection of stanozolol and methanedienone causes a rapid cytosolic depletion of AR in rat skeletal muscle. Based on these results, we conclude that anabolic steroids with low affinity to AR in vitro, can in fact in vivo act on the AR to cause biological responses. PMID- 15876414 TI - Induction of hepatic drug-metabolising enzymes and tamoxifen metabolite profile in relation to administration route during low-dose treatment in nude rats. AB - Tamoxifen is the most used anticancer drug and is approved for chemoprevention. Little is known about the enzyme inducing properties of low-dose regimens and the influence of route of administration. In this study, nude rats received 5 mg/kg/day of tamoxifen orally or a 50 mg continuous-release pellet subcutaneously. The mRNAs for cytochrome P450-enzymes (CYPs), flavin-containing monooxygenase 1 (FMO1) and phase II drug-metabolising enzymes were quantified by real-time RT-PCR. Tamoxifen and metabolite concentrations were measured using HPLC. We observed a significant increase in CYP3A18 and FMO1 mRNA expression levels in the orally treated animals, whereas the increase in CYP3A2 expression did not reach statistical significance (p=0.057). No significant induction of enzyme expression was observed in rats that received subcutaneous (S.c.) treatment. After 33 days the serum levels of 4-hydroxytamoxifen (4OHtam), tamoxifen and N-desmethyltamoxifen (NDtam) in orally treated animals were 1.8+/ 0.7, 11.1+/-3.2 and 11.4+/-3.8 ng/ml, respectively. In subcutaneously treated animals, tamoxifen and N-desmethyltamoxifen were detected in tissues, but not in serum. These data demonstrate that in contrast to the subcutaneous administration, low-dose oral tamoxifen induced tamoxifen-metabolising enzymes. Furthermore, the different routes of administration resulted in different serum and tissue levels of tamoxifen and metabolites. PMID- 15876415 TI - Phytoestrogens derived from red clover: an alternative to estrogen replacement therapy? AB - The benefits of plant extracts from soy and red clover as alternatives to conventional hormone replacement therapy (HRT) have been debated in the past. Here, an attempt has been made to summarize the biochemical and pharmacological data in the light of clinical aspects. Red clover and soy extracts contain isoflavones, which have a high affinity to estrogen receptor alpha (ERalpha), estrogen receptor beta (ERbeta), progesterone receptor (PR) and androgen receptor (AR). The higher affinity to ERbeta compared to ERalpha has been used as an explanation why red clover extracts function as food additives to treat menopausal disorders and may reduce risk of breast cancer. Biochemical analysis shows that these representatives of phytoestrogens have multiple actions beside selective estrogen receptor modulator (SERM)-activity. They act as selective estrogen enzyme modulators (SEEMs), have antioxidant activity and interact with transcription factors such as NF-kappaB. Furthermore, it is indicated that they have protective effects on osteoporosis and the cardiovascular system. Currently 40-50mg of isoflavones (biochanin A, daidzein, formononetin and genistein) are recommended as daily dose. This recommendation is based on the daily intake of phytoestrogens in a traditional Japanese diet. PMID- 15876416 TI - Localization of 17beta-hydroxysteroid dehydrogenase in Mycobacterium sp. VKM Ac 1815D mutant strain. AB - The localization of mycobacterial 17beta-hydroxysteroid dehydrogenase (17beta-OH SDH) was studied using cell fractionation and cytochemical investigation. Mycobacterium sp. Et1 mutant strain derived from Mycobacterium sp. VKM Ac-1815D and characterized by increased 17beta-OH SDH activity was used as a model organism. Subcellular distribution study showed both soluble and membrane-bound forms of mycobacterial 17beta-hydroxysteroid dehydrogenase. The cytochemical method based on a copper ferrocyanide procedure followed by electron microscopic visualization was applied in order to investigate the intracellular localization of bacterial 17beta-OH SDH in more detail. The enzyme was found to be located in the peripheral cytoplasmic zone adjoining the cytoplasmic membrane (CM). 17beta OH SDH was loosely membrane bound and easily released into the environment under the cell integrity failure. PMID- 15876417 TI - Are speed enforcement cameras more effective than other speed management measures? An evaluation of the relationship between speed and accident reductions. AB - In this paper, models are developed which enable a prediction of how the impact of speed management schemes on accidents varies both with speed changes and with site and scheme characteristics. It was found that, the impact of schemes with vertical deflections is independent of the change in mean speed: an accident reduction of 44% is predicted by the model irrespective of the impact on speed. For cameras and other types of engineering schemes, a simple relationship between the change in mean speed and the consequent change in accidents is available. For the range of mean speeds typically found on 30 mph roads, the percentage accident reduction per 1 mph speed reduction is around 4% for cameras and 7-8% for schemes with horizontal features. While larger percentage accident reductions are achieved per 1 mph speed reduction on lower speed roads, larger speed reductions and larger overall percentage accident reductions are obtained on roads with higher before mean speeds. It is possible to predict both changes in speeds and accidents before treatment using the models derived from this study and these models confirm that schemes with vertical deflections are most effective in reducing both speeds and accidents. PMID- 15876418 TI - Perception of successive targets presented in invariant-item streams. AB - When two successive, spatially overlapping, targets (S1 and S2) are presented on a blank background, S2 typically dominates in explicit perception. We tested whether S2 dominance is also found for the conditions of presenting S1 and S2 in a stream of irrelevant objects. Successive target letters were presented within a stream of invariant stimulus items (capital Is). The stream items were presented either as a perceptually continuous object where both type and token appeared invariant (60-Hz stream) or as a flickering stream of successive replicas of the invariant stationary object where the type appeared invariant but the token appearance seemed intermittent (20-Hz condition). Compared to the control condition where targets were presented on a blank background we found that (1) recognition rate was lower for targets embedded in a perceptually continuous type and-token object (60 Hz), but was unchanged for targets in a perceptually flickering sequence of the invariant-object tokens (20 Hz); (2) S1 recognition rate was higher compared to S2 recognition rate within the first epoch of stream (0-150 ms) while within the later stream-epochs S2 dominated over S1 as usual; (3) the overall difference in recognition rates between S1 and S2 was decreased. The results are discussed in the theoretical context of visual masking and attentional blink. PMID- 15876419 TI - Conjunction errors in recognition memory: modality-free errors for older adults but not for young adults. AB - A dual-process theory of memory was applied to processes in normal aging, with a focus on recognition errors in the feature-conjunction paradigm (i.e., false recognition of blackbird after studying parent words blackmail and/or jailbird). Study repetition was manipulated so that some parent words occurred once and others occurred three times. Age-related differences on hit scores occurred for two experiments. The results for feature and conjunction conditions showed repetition effects but no age-related differences when participants were uninformed of the lures (Experiment 1). However, age-related differences emerged when the retrieval of modality source information created a way to evade conjunction errors (Experiment 2). In the second experiment, study repetition decreased errors for the young adults but increased errors for the older adults, and young adults were better able than older adults to avoid conjunction errors when the parent words had been repeated. For older adults, the conjunction errors were modality-free. The results provide additional evidence that older adults experience difficulty in recollecting aspects of a study experience, and the results from groups of young adults required to respond quickly on the tests provide converging evidence for this conclusion. PMID- 15876420 TI - Contribution of the pfmdr1 gene to antimalarial drug-resistance. AB - The emergence of drug-resistance poses a major obstacle to the control of malaria. A homolog of the major multidrug-transporter in mammalian cells was identified, Plasmodium falciparum multidrug resistance protein-1, pfmdr1, also known as the P-glycoprotein homolog 1, Pgh-1. Several studies have demonstrated strong, although incomplete, associations between resistance to the widely used antimalarial drug chloroquine and mutation of the pfmdr1 gene in both laboratory and field isolates. Genetic studies have confirmed a link between mutation of the pfmdr1 gene and chloroquine-resistance. Although not essential for chloroquine resistance, pfmdr1 plays a role in modulating levels of resistance. At the same time it appears to be a significant component in resistance to the structurally related drug quinine. A strong association has been observed between possession of the wildtype form of pfmdr1, amplification of pfmdr1 and resistance to hydrophobic drugs such as the arylaminoalcohol mefloquine and the endoperoxide artemisinin derivatives in field isolates. This is supported by genetic studies. The arylaminoalcohol and endoperoxide drugs are structurally unrelated drugs and this resistance resembles true multidrug resistance. Polymorphism in pfmdr1 and gene amplification has been observed throughout the world and their usefulness in predicting resistance levels is influenced by the history of drug selection of each population. PMID- 15876421 TI - Serine proteinase over-expression in relation to deltamethrin resistance in Culex pipiens pallens. AB - Two serine proteinase genes were isolated from Culex pipiens pallens as significantly up-regulated genes in a deltamethrin-resistant strain through a combination of suppression substractive hybridization and gene expression profiling by macroarrays. These two genes were found to be expressed at least threefold higher in the resistant strain than in the susceptible one. By using rapid amplification of cDNA ends to screen the constructed cDNA library, we cloned these two sequences. There were 909 bp with an open reading frame of 786 bp in the sequence of trypsin cDNA (GenBank/NCBI AF468495), the deduced protein had 261 amino acids, which was most similar to the trypsin gene of Anopheles gambiae. There were 992 bp with an open reading frame of 816 bp in the chymotrypsin cDNA (GenBank/NCBI AY034060), and its deduced amino acid sequence had 271 amino acids, which was most similar to the chymotrypsin-like protein from Aedes aegypti. The two genes were stably expressed in mosquito C6/36 cells, and the expected 29 and 30 kDa bands were shown with Western blot, respectively. In these cells, after deltamethrin treatment, they had protective effects on the viability. The results indicate that trypsin and chymotrypsin were more highly expressed in the deltamethrin-resistant strain, and was related to insecticide resistance in mosquitoes, Cx. pipiens pallens. PMID- 15876422 TI - Formation and disposition of diethylphosphoryl-obidoxime, a potent anticholinesterase that is hydrolyzed by human paraoxonase (PON1). AB - The potential of pyridinium-4-aldoximes, such as obidoxime, to reactivate diethylphosphorylated acetylcholinesterases is not fully exploited due to the inevitable formation of phosphoryloximes (POX) with high anticholinesterase activity. Mono(diethylphosphoryl) obidoxime (DEP-obidoxime) was isolated for the first time showing remarkable stability under physiological conditions (half-life 13.5min; pH 7.1; 37 degrees C). The half-life was considerably extended to 20h at 0 degrees C, which facilitated the preparation and allowed isolation by HPLC. The structure was confirmed by mass spectrometry and the degradation pattern. DEP obidoxime decomposed by an elimination reaction forming the intermediate nitrile that hydrolyzed mainly into the pyridone and cyanide. The intermediates were prepared and confirmed by mass spectroscopy. DEP-Obidoxime was an extremely potent inhibitor of human acetylcholinesterase approaching a second-order rate constant of 10(9)M(-1)min(-1) (pH 7.4; 37 degrees C). The nitrile and the pyridone were still good reactivators. In the presence of human plasma DEP obidoxime was hydrolyzed into parent obidoxime. Calcium-dependence and sensitivity towards chelators, substitution pattern by other divalent cations and protein-modifying agents all pointed to human paraoxonase (hPON1) as the responsible protein with POX-hydrolase activity. Subjects, probably belonging to the homozygous (192)arginine subtype, were virtually devoid of POX-hydrolase activity while a highly purified hPON1 of the homozygous (192)glutamine subtype exhibited particularly high POX-hydrolase activity. Two parathion-poisoned patients with high and low POX-hydrolase activity responded well and poorly, respectively, to obidoxime treatment although the former patient had higher plasma paraoxon levels than the poor responder. Hence, the POX-hydrolase associated PON1 subtype may be another contributor that modulates pyridinium-4 aldoxime effectiveness. PMID- 15876423 TI - Quercetin, but not rutin and quercitrin, prevention of H2O2-induced apoptosis via anti-oxidant activity and heme oxygenase 1 gene expression in macrophages. AB - In the present study, we examine the protective mechanism of quercetin (QE) on oxidative stress-induced cytotoxic effect in RAW264.7 macrophages. Results of Western blotting show that QE but not its glycoside rutin (RUT) and quicitrin induced HO-1 protein expression in a time- and dose-dependent manner, and HO-1 protein induced by QE was blocked by an addition of cycloheximide or actinomycin D. Induction of HO-1 gene expression by QE was accompanied by inducing ERKs, but not JNKs or p38, proteins phosphorylation. Addition of PD98059, but not SB203580 or SP600125, significantly attenuates QE-induced HO-1 protein and mRNA expression associated with blocking the expression of phosphorylated ERKs proteins. H(2)O(2) addition reduces the viability of cells by MTT assay, and appearance of DNA ladders, hypodiploid cells, and an increase in intracellular peroxide level was detected. Addition of QE, but not QI or RUT, significantly reduced the cytotoxic effect induced by H(2)O(2) associated with blocking the production of intracellular peroxide, DNA ladders, and hypodiploid cells. QE protection of cells from H(2)O(2)-induced apoptosis was significantly suppressed by adding HO inhibitor SnPP or ERKs inhibitor PD98059. Additionally, QE protects cells from H(2)O(2)-induced a decrease in the mitochondrial membrane potential and a release of cytochrome c from mitochondria to cytosol by DiOC6 and Western blotting assay, respectively. Activation of apoptotic proteins including the caspase 3, caspase 9, PARP, D4-GDI proteins was identified in H(2)O(2)-treated cells by Western blotting and enzyme activity assay, and that was significantly blocked by an addition of QE, but not RUT and QI. Furthermore, HO-1 catalytic metabolites carbon monoxide (CO), but not Fe(2+), Fe(3+), biliverdin or bilirubin, performed protective effect on cells from H(2)O(2)-induced cell death with an increase in HO-1 protein expression and ERKs protein phosphorylation. These data suggest that induction of HO-1 protein may participate in the protective mechanism of QE on oxidative stress (H(2)O(2))-induced apoptosis, and reduction of intracellular ROS production and mitochondria dysfunction with blocking apoptotic events were involved. Differential anti-apoptotic effect between QE and its glycosides RUT and QI via distinct HO-1 protein induction was also delineated. PMID- 15876424 TI - Role of P-glycoprotein in transplacental transfer of methadone. AB - Methadone is the therapeutic agent of choice for treatment of the pregnant opiate addict. However, little is known on the factors affecting its concentration in the fetal circulation during pregnancy and how it might relate to neonatal outcome. Therefore, a better understanding of the function of placental metabolic enzymes and transporters should add to the knowledge of the role of the tissue in the disposition of methadone and its relation to neonatal outcome. We hypothesized that the expression and activity of the placental efflux transporter P-glycoprotein (P-gp) would affect the transfer of methadone to the fetal circulation. Data obtained utilizing dual perfusion of placental lobule and monolayers of Be-Wo cell line indicated that methadone is extruded by P-gp. Transfer of methadone to the fetal circuit was increased by 30% in the presence of the P-gp inhibitor GF120918 while the transfer of paclitaxel, a typical substrate of the glycoprotein, was increased by 50%. In the Be-Wo cell line, methadone and paclitaxel uptake was also increased in the presence of the P-gp inhibitor cyclosporin A. Moreover, the expression of P-gp in placental brush border membranes varied between term placentas. Taken together, these data strongly suggest that the concentration of methadone in the fetal circulation is affected by the expression and activity of P-gp. It is reasonable to speculate that placental disposition of methadone affects its concentration in the fetal circulation. If true, this may also be directly related to the incidence and intensity of neonatal abstinence syndrome (NAS). PMID- 15876425 TI - Rosiglitazone, an agonist of peroxisome proliferator-activated receptor gamma, reduces chronic colonic inflammation in rats. AB - Recent studies have shown the implication of the peroxisome proliferator activated receptor gamma (PPARgamma) in control of inflammation, immune and apoptotic responses during early experimental colitis. However, there is little information about the effects of these agents on colonic mucosa under chronic inflammatory conditions. In this study, we have evaluated the effects of rosiglitazone, a PPAR-gamma agonist, on the chronic injury caused by intra colonic administration of trinitrobenzensulfonic acid (TNBS) in rats. Rosiglitazone (1 and 5mg/kg p.o.) was administered by oral gavage, 24h after TNBS instillation and daily during 2 weeks before killing the rats. Colons were removed for histological and biochemical analysis. Administration of rosiglitazone corrected the disorders in morphology associated to lesions, significantly reduced the ulceration index, the rise of myeloperoxidase (MPO) and the levels of tumour necrosis factor alpha (TNF-alpha). In addition, rosiglitazone treatment increased prostaglandin (PG)E(2) production and returned PGD(2) to basal levels. Also, reduced cyclooxygenase (COX)-2 and nuclear transcription factor NF-kappa B (NF-kappaB) p65 proteins expression. Furthermore, treatment of rats with rosiglitazone caused a significant increase of TNBS induced apoptosis. In summary, rosiglitazone exerts protective effects in chronic experimental colitis. The anti-inflammatory effects seem to be related to impairment of neutrophil function, absence of up-regulation of TNF-alpha and decrease of nuclear NF-kappaB p65 expression. Our results also suggest that the activation of the PPARgamma pathway reduces COX-2 overexpression, returns the increased PGD(2) values to basal levels and induces a significant increase of TNBS-induced apoptosis. We conclude that rosiglitazone represents a novel approach to the treatment of ulcerative colitis. PMID- 15876426 TI - Constitutive expression of CIITA directs CD4 T cells to produce Th2 cytokines in the thymus. AB - We generated mice expressing a human type III CIITA transgene (CIITA Tg) under control of the CD4 promoter to study the role of CIITA in CD4 T cell biology. The transgene is expressed in peripheral CD4 and CD8 T cells, as well as in thymocytes. When CD4 T cells were differentiated towards the Th2 lineage, both control and CIITA Tg Th2 cells expressed similar levels of Th2 cytokines. Th1 cells from control and CIITA Tg mice cells produced comparable levels of IFN gamma. CIITA Tg Th1 cells also expressed IL-4, IL-5, and IL-13 in the absence of Stat6. There was an approximate 10-fold increase in the number of peripheral naive CD4 T cells and NK1.1- thymocytes producing IL-4 from CIITA Tg mice compared to control mice. Finally, Th1 cells from irradiated control mice reconstituted with CIITA Tg bone marrow displayed the same cytokine production profiles as Th1 cells from CIITA Tg mice. Together, our data demonstrate that CIITA expression pre-disposes CD4 T cells to produce Th2 type cytokines. Moreover, phenotypic similarities between Th1 cells expressing the CIITA transgene and CIITA deficient Th1 cells suggest that the role of CIITA in cytokine regulation is complex and may reflect both direct and indirect mechanisms of T cell development and differentiation. PMID- 15876427 TI - The mouse immune response to human fibrinogen reveals an autoimmune component against mouse fibrinogen. AB - The present experiments were carried out to analyze whether immunization of mice with human fibrinogen would induce autoimmunity like other heterologous proteins such as collagen type II, thyroglobulin or myelin basic protein. Our results demonstrate that human fibrinogen induces very strong immune responses in all mouse strains analyzed. Autoimmune responses with short-term memory to mouse fibrinogen are induced in genetically susceptible mice. These autoimmune Th2-type responses induce splenomegaly, enhanced coagulation times, and production of rheumatoid factors. The short-lived autoimmune memory was not regulated by either suppressor T cells or exhaustion of immune cells; rather this potentially dangerous autoimmune response was regulated by unknown, antigen-specific feedback mechanisms (they do not influence immune responses to proteins like HSA and OA in the same mice). Such feedback mechanisms were not found in the immune responses to other heterologous proteins inducing significant cross-reactive autoimmunity such as collagen type II, thyroglobulin, or myelin basic protein. PMID- 15876428 TI - Lipopolysaccharide negatively modulates vitamin D action by down-regulating expression of vitamin D-induced VDR in human monocytic THP-1 cells. AB - Vitamin D3, an important seco-steroid hormone for the regulation of body calcium homeostasis, promotes immature myeloid precursor cells to differentiate into monocytes/macrophages. Vitamin D receptor (VDR) belongs to a nuclear receptor super-family that mediates the genomic actions of vitamin D3 and regulates gene expression by binding with vitamin D response elements in the promoter region of the cognate gene. Thus by regulating gene expression, VDR plays an important role in modulating cellular events such as differentiation, apoptosis, and growth. Here we report lipopolysaccharide (LPS), a bacterial toxin; decreases VDR protein levels and thus inhibits VDR functions in the human blood monocytic cell line, THP-1. The biologically active form of vitamin D3, 1alpha,25-dihydroxy vitamin D3 [1,25(OH)2D3], induced VDR in THP-1 cells after 24 h treatment, and LPS inhibited 1,25(OH)2D3-mediated VDR induction. However, LPS and 1,25(OH)2D3 both increased VDR mRNA levels in THP-1 cells 20 h after treatment, as observed by real time RT PCR. Moreover, LPS plus 1,25(OH)2D3 action on VDR mRNA level was additive and synergistic. A time course experiment up to 60 h showed an increase in VDR mRNA that was not preceded with an increase in VDR protein levels. Although the proteasome pathway plays an important role in VDR degradation, the proteasome inhibitor lactacystin had no effect on the LPS-mediated down-regulation of 1,25(OH)2D3 induced VDR levels. Reduced VDR levels by LPS were accompanied by decreased 1,25(OH)2D3/VDR function determined by VDR responsive 24-hydroxylase (CYP24) gene expression. The above results suggest that LPS impairs 1,25(OH)2D3/VDR functions, which may negatively affect the ability of 1,25(OH)2D3 to induce myeloid differentiation into monocytes/macrophages. PMID- 15876429 TI - Non-symbolic arithmetic in adults and young children. AB - Five experiments investigated whether adults and preschool children can perform simple arithmetic calculations on non-symbolic numerosities. Previous research has demonstrated that human adults, human infants, and non-human animals can process numerical quantities through approximate representations of their magnitudes. Here we consider whether these non-symbolic numerical representations might serve as a building block of uniquely human, learned mathematics. Both adults and children with no training in arithmetic successfully performed approximate arithmetic on large sets of elements. Success at these tasks did not depend on non-numerical continuous quantities, modality-specific quantity information, the adoption of alternative non-arithmetic strategies, or learned symbolic arithmetic knowledge. Abstract numerical quantity representations therefore are computationally functional and may provide a foundation for formal mathematics. PMID- 15876430 TI - Peroxiredoxin-I is an autoimmunogenic tumor antigen in non-small cell lung cancer. AB - In eukaryotic cells, peroxiredoxins are both antioxidants and regulators of H(2)O(2)-mediated signaling. We previously found that peroxiredoxin-I (Prx-I) was overexpressed in non-small cell lung cancer (NSCLC) tissue. Since overexpressed protein can induce a humoral immune response, we examined whether serum from NSCLC patients exhibited immunoreactivity against Prx-I using Western blotting. We found that 25 (47%) of 53 NSCLC patients tested had autoantibodies against Prx I in their sera, whereas such activity was detected in 4 (8%) sera from 50 healthy subjects. Prx-I itself was detected in the sera from 18 (34%) of 53 NSCLC patients but in only 1 (2%) serum from 50 controls. Moreover, 17% of NSCLC sera were positive to both Prx-I antibody and antigen but none in control sera. The data indicate both Prx-I autoantibody and circulating antigen are potential biomarkers for use in serological diagnosis of NSCLC. Interestingly enough, we found that Prx-I was secreted by lung adenocarcinoma cells (A549) but not by non cancer lung cells (BEAS 2B) or breast cancer cells (MCF7). This cell culture study suggests the possibility of Prx-I secretion from NSCLC tumor tissues. PMID- 15876431 TI - The longin domain regulates subcellular targeting of VAMP7 in Arabidopsis thaliana. AB - SNAREs (soluble N-ethyl-maleimide sensitive factor attachment protein receptors) which locate on the specific organelle membrane assure the correct vesicular transport by mediating specific membrane fusions. SNAREs are referred to as R- or Q-SNAREs on the basis of the amino acid sequence similarities and specific conserved residues. All of the Arabidopsis R-SNAREs have a N-terminal domain, called the longin domain (LD). In this study, we investigated the vacuolar targeting mechanism of Arabidopsis R-SNAREs. The vacuolar localized AtVAMP711 was used as the mother protein of GFP-tagged chimeric proteins joined to several domains such as the LD, the SNARE motif (SNM) and the transmembrane domain (TMD) of other organelle-localized R-SNAREs. The results showed that, whereas the TMD is not relevant for the vacuolar targeting, a complete LD is essential for the vacuolar and subcellular targeting. PMID- 15876432 TI - An NMR-based docking model for the physiological transient complex between cytochrome f and cytochrome c6. AB - The physiological transient complex between cytochrome f (Cf) and cytochrome c(6) (Cc(6)) from the cyanobacterium Nostoc sp. PCC 7119 has been analysed by NMR spectroscopy. The binding constant at low ionic strength is 8 +/- 2 mM(-1), and the binding site of Cc(6) for Cf is localized around its exposed haem edge. On the basis of the experimental data, the resulting docking simulations suggest that Cc(6) binds to Cf in a fashion that is analogous to that of plastocyanin but differs between prokaryotes and eukaryotes. PMID- 15876433 TI - Effects of lipid kinase expression and cellular stimuli on phosphatidylinositol 5 phosphate levels in mammalian cell lines. AB - Phosphatidylinositol 5-phosphate (PtdIns5P) is a relatively recently discovered inositol lipid whose metabolism and functions are not yet clearly understood. We have transfected cells with a number of enzymes that are potentially implicated in the synthesis or metabolism of PtdIns5P, or subjected cells to a variety of stimuli, and then measured cellular PtdIns5P levels by a specific mass assay. Stable or transient overexpression of Type IIalpha PtdInsP kinase, or transient overexpression of Type Ialpha or IIbeta PtdInsP kinases caused no significant change in cellular PtdIns5P levels. Similarly, subjecting cells to oxidative stress or EGF stimulation had no significant effect on PtdIns5P, but stimulation of HeLa cells with a phosphoinositide-specific PLC-coupled agonist, histamine, caused a 40% decrease within 1 min. Our data question the degree to which inositide kinases regulate PtdIns5P levels in cells, and we discuss the possibility that a significant part of both the synthesis and removal of this lipid may be regulated by phosphatases and possibly phospholipases. PMID- 15876434 TI - Adaptation of Saccharomyces cerevisiae to high hydrostatic pressure causing growth inhibition. AB - Genome-wide mRNA expression profiles of Saccharomyces cerevisiae growing under hydrostatic pressure were characterized. We selected a hydrostatic pressure of 30 MPa at 25 degrees C because yeast cells were able to grow under these conditions, while cell size and complexity were increased after decompression. Functional characterization of pressure-induced genes suggests that genes involved in protein metabolism and membrane metabolism were induced. The response to 30 MPa was significantly different from that observed under lethal conditions because protein degradation was not activated under 30 MPa pressure. Strongly induced genes those that contribute to membrane metabolism and which are also induced by detergents, oils, and membrane stabilizers. PMID- 15876435 TI - Adaptor usage and Toll-like receptor signaling specificity. AB - It is now well established that Toll-like receptors (TLRs) act as primary sensors of microbial compounds. Details of the molecular mechanisms governing TLR responses are emerging steadily and our understanding of the signaling pathways activated these receptors has improved greatly over the last few years. Differences in adaptor usage, cellular localisation and signaling cascades have been elucidated. In this review we will summarize the current understanding of TLR signaling and its regulation. PMID- 15876436 TI - Newer aminopyrimidinimino isatin analogues as non-nucleoside HIV-1 reverse transcriptase inhibitors for HIV and other opportunistic infections of AIDS: design, synthesis and biological evaluation. AB - Human immuno deficiency virus (HIV) weakens the immune system so that many opportunistic infections (OIs) like tuberculosis, hepatitis, bacterial infections etc can develop. In this paper, we designed aminopyrimidinimino isatin lead compound as a novel non-nucleoside reverse transcriptase inhibitor (NNRTI) with broad-spectrum chemotherapeutic properties for the effective treatment of AIDS and AIDS-related OIs. Compound 1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-[[N4 [3'-(4'-amino-5'-trimethoxybenzyl pyrimidin-2'-yl)imino-1'-(5 methylisatinyl)]methyl]-N1-piperazinyl]-3-quinoline carboxylic acid (10) emerged as the most potent broad-spectrum chemotherapeutic agent active against HIV, HCV, Mycobacterium tuberculosis and various pathogenic bacteria. PMID- 15876437 TI - Karuppasamy K, Hutton MJ, LeF Porteous MJ. A novel system for the irrigation of open fractures [Injury 35 (10) (2004) 980-981]. PMID- 15876438 TI - Genotypic and phenotypic diversity of a baculovirus population within an individual insect host. AB - It is becoming increasingly apparent that many pathogen populations, including those of insects, show high levels of genotypic variation. Baculoviruses are known to be highly variable, with isolates collected from the same species in different geographical locations frequently showing genetic variation and differences in their biology. More recent studies at smaller scales have also shown that virus DNA profiles from individual larvae can show polymorphisms within and between populations of the same species. Here, we investigate the genotypic and phenotypic variation of an insect baculovirus infection within a single insect host. Twenty four genotypically distinct nucleopolyhedrovirus (NPV) variants were isolated from an individual pine beauty moth, Panolis flammea, caterpillar by in vivo cloning techniques. No variant appeared to be dominant in the population. The PaflNPV variants have been mapped using three restriction endonucleases and shown to contain three hypervariable regions containing insertions of 70-750 bp. Comparison of seven of these variants in an alternative host, Mamestra brassicae, demonstrated that the variants differed significantly in both pathogenicity and speed of kill. The generation and maintenance of pathogen heterogeneity are discussed. PMID- 15876439 TI - Saturated salt solutions for humidity control and the survival of dry powder and oil formulations of Beauveria bassiana conidia. AB - Oil-based formulated conidia sprayed on steel plates and conidia powder (control) of Beauveria bassiana isolate IMI 386243 were stored at temperatures from 10 to 40 degrees C in desiccators over saturated salt solutions providing relative humidities from 32 to 88%, or in hermetic storage at 40 degrees C, and moisture contents in equilibrium with 33 or 77% relative humidity. The negative semi logarithmic relation (P<0.005) between conidia longevity (at 40 degrees C) and equilibrium relative humidity did not differ (P>0.25) between formulated conidia and conidia powder. Despite this, certain saturated salts provided consistently greater longevity (NaCl) and others consistently shorter longevity (KCl) for formulated conidia compared to conidia powder. These results, analysis of previous data, and comparison with hermetic storage, indicate that storage of conidia over saturated salt solutions provides inconsistent responses to environment and so may be problematic for bio-pesticide research. In hermetic storage, oil formulation was not deleterious to longevity and in the more moist environment enhanced survival periods. PMID- 15876440 TI - Restitution of visual functions in cerebrally blind children. AB - In adult patients who suffer from a visual field defect due to cerebral lesions, visual functions can be restored by systematic visual field training. Such visual field training is not feasible in young, brain-damaged children, who are unable to cooperate like adults. We have already shown earlier [Werth, R., and Moehrenschlager, M. (1999). The development of visual functions in cerebrally blind children during a systematic visual field training. Restorative Neurology and Neuroscience, 15, 229-241.] that systematic visual field training, which requires no ability to understand instructions or to cooperate, is successful in brain-damaged children and may lead to complete recovery of the visual field within 3 months. The present study provides more behavioral evidence and a control of intraocular light scatter. In addition to the earlier study, the luminance difference thresholds in the recovered visual field were compared with those of a normal control group. Seventeen children aged 1-4 years who had been blind for more than 1 year after perinatal asphyxia and two children suffering from homonymous hemianopia due to asphyxic-ischemic lesions participated in systematic visual field training. The functional visual field was assessed with a specially designed arc perimeter. Visual functions developed within a training period of 3 months in 11 children who received visual field training, whereas there was no recovery in the control group (N=37). The findings support the assumption that systematic visual field training facilitates the development of vision in cerebrally blind children. In two children who recovered from blindness, it was shown in functional magnetic resonance imaging (fMRI) that brain tissue in the area of the visual cortex contralateral to the blind visual hemifield was activated by light. In two children suffering from asphyxic ischemic lesions who did not recover only brain tissue in the area of the visual cortex contralateral to the good visual hemifield could be activated by light. These results support the assumption that activity in spared tissue of the striate and extrastriate visual cortex are a necessary condition for recovery of the visual field in children suffering from cerebral blindness. PMID- 15876441 TI - Brainstem auditory evoked potentials and flash visual evoked potentials in Vietnamese miniature pot-bellied pigs. AB - Brainstem auditory evoked potentials (BAEP) and flash visual evoked potentials (VEP) were recorded from juvenile (5-7 weeks of age) and adult Vietnamese miniature pot-bellied pigs to provide normative data for clinical applications. BAEP responses were collected in response to stimulus intensities of 85, 95, and 105 dB nHL. VEP responses were collected in response to flashes of white light in a darkened room. Left-right ear and left-right eye responses did not differ significantly, and were combined for analysis, with each animal providing two data points for each response. BAEP responses in juvenile subjects were mature, and in all subjects showed the typical pattern of decreasing peak latencies and increasing peak amplitudes with increasing stimulus intensity. VEP responses in juvenile subjects were near to mature values, but the latencies still exceeded those of adults. Differences in response maturation between precocial and altricial species are discussed. PMID- 15876442 TI - Stage-dependent production and release of histidine-rich protein 2 by Plasmodium falciparum. AB - Because of their sequestration in the microcirculation, the pathogenic late stages of Plasmodium falciparum are under-represented in peripheral blood samples from patients with falciparum malaria. Excreted products of the parasite might help to estimate this sequestered biomass. We quantified the stage-dependent production and release per parasite of P. falciparum histidine-rich protein 2 (PfHRP2) with the objective of measuring the sequestered biomass. A simple method to relate parasite stage to parasite age was developed to facilitate this. In four isolates of P. falciparum, the median (range) PfHRP2 content was 2.0fg (0.5 4.3fg) for a young ring stage infected erythrocyte, and 5.4fg (2.1-10.2fg) for the schizont stage. The amount of PfHRP2 in the parasitized erythrocyte increased most during development to the mature trophozoite stage. The median (range) amount of PfHRP2 secreted per parasite per entire erythrocytic cycle was 5.2fg (1.1-13.0fg). A median of 89% of the total PfHRP2 was excreted at the moment of schizont rupture. This assessment of the stage-dependent release of PfHRP2 is an essential prerequisite for future studies aimed at estimating the total patient parasite mass from the peripheral blood PfHRP2 concentration. PMID- 15876443 TI - Antimalarial efficacy of chloroquine, amodiaquine, sulfadoxine-pyrimethamine, and the combinations of amodiaquine + artesunate and sulfadoxine-pyrimethamine + artesunate in Huambo and Bie provinces, central Angola. AB - We studied three antimalarial treatments in Caala and Kuito, Angola, in 2002 and 2003. We tested chloroquine (CQ), amodiaquine (AQ) and sulfadoxine-pyrimethamine (SP) in Caala, and AQ, SP and the combinations AQ+artesunate (AQ+AS) and SP+artesunate (SP+AS) in Kuito. A total of 619 children (240 in Caala, 379 in Kuito) with uncomplicated Plasmodium falciparum malaria were followed-up for 28 days, with PCR genotyping to distinguish recrudescence from reinfection. PCR corrected failure proportions at day 28 were very high in the CQ group (83.5%, 95% CI 74.1-90.5), high in the SP groups (Caala: 25.3%, 95% CI 16.7-35.8; Kuito: 38.8%, 95% CI 28.4-50.0), around 20% in the AQ groups (Caala: 17.3%, 95% CI 10.0 27.2; Kuito: 21.6%, 95% CI 14.3-30.6) and very low in the artemisinin-based combination groups (1.2%, 95% CI 0.0-6.4 for each combination AQ+AS and SP+AS). These results show that CQ and SP are no longer efficacious in Caala and Kuito and that the moderate efficacy of AQ is likely to be compromised in the short term if used as monotherapy. We recommend the use of AQ with AS, though this combination might not have a long useful therapeutic life because of AQ resistance. PMID- 15876444 TI - Identification of novel bradykinin-potentiating peptides and C-type natriuretic peptide from Lachesis muta venom. AB - The generation of expressed sequence tags (ESTs) from the pit-viper snake Lachesis muta venom glands allowed us to identify two cDNA isoforms which encode the precursors for bradykinin-potentiating peptides (BPPs) and a C-type natriuretic peptide (CNP). The sequence data derived from these cDNAs combined with the venom peptides identification using MALDI-TOF mass spectrometry analysis predicted that these molecules are the precursor protein isoforms that are further processed to produce five novel BPPs and a CNP. They were identified directly in crude venom using MALDI-TOF. The BPPs sequences were further confirmed by MALDI-TOF/TOF de novo sequencing, and an unusual BPP with a residue of tryptophan at the N-terminus (usually it is pyroglutamate) was identified. The putative processing steps required to form the mature BPPs and CNP seem to be similar to those proposed for the ones found in the venom of Bothrops jararaca and Glodyus blomhoffi. PMID- 15876445 TI - Snake venom C-type lectins interacting with platelet receptors. Structure function relationships and effects on haemostasis. AB - Snake venoms contain components that affect the prey either by neurotoxic or haemorrhagic effects. The latter category affect haemostasis either by inhibiting or activating platelets or coagulation factors. They fall into several types based upon structure and mode of action. A major class is the snake C-type lectins or C-type lectin-like family which shows a typical folding like that in classic C-type lectins such as the selectins and mannose-binding proteins. Those in snake venoms are mostly based on a heterodimeric structure with two subunits alpha and beta, which are often oligomerized to form larger molecules. Simple heterodimeric members of this family have been shown to inhibit platelet functions by binding to GPIb but others activate platelets via the same receptor. Some that act via GPIb do so by inducing von Willebrand factor to bind to it. Another series of snake C-type lectins activate platelets by binding to GPVI while yet another series uses the integrin alpha(2)beta(1) to affect platelet function. The structure of more and more of these C-type lectins have now been, and are being, determined, often together with their ligands, casting light on binding sites and mechanisms. In addition, it is relatively easy to model the structure of the C-type lectins if the primary structure is known. These studies have shown that these proteins are quite a complex group, often with more than one platelet receptor as ligand and although superficially some appear to act as inhibitors, in fact most function by inducing thrombocytopenia by various routes. The relationship between structure and function in this group of venom proteins will be discussed. PMID- 15876446 TI - Molecular cloning of serine proteinases from Bothrops jararaca venom gland. AB - Snake venom is known to contain an abundance of enzyme isoforms, and various disorders associated with envenomation have been ascribed partially to their diversified functions. Crude venom of Bothrops jararaca was subjected to conventional two-dimensional SDS-PAGE, followed by immunoblot analysis using an antiserum raised against KN-BJ 2, a serine proteinase previously isolated from this venom. A number of immunoreactive proteins with comparable molecular masses and different pIs emerged, implying the venom contains yet-unknown serine proteinases. A B. jararaca venom gland cDNA library was subsequently screened with a labeled KN-BJ 2 cDNA as a probe. Among a number of positive cDNA clones, three--HS112, HS114, and HS120--were selected and sequenced. These clones each had an open reading frame of 759-774 bp, and their deduced amino acid sequences illustrated considerable similarities to that of KN-BJ 2 as well as to those of serine proteinases of different origins. However, no apparent match to any of the deposited sequences was found in the current GenBank/EMBL databases, indicating that each of these cDNA clones encodes a serine proteinase distinct from the known enzymes. Analyses of the nucleotide and amino acid sequences of these cDNA clones support the accelerated evolution hypothesis proposed for snake venom enzymes. PMID- 15876447 TI - Illusory rebound motion and the motion continuity heuristic. AB - A new motion illusion, "illusory rebound motion" (IRM), is described. IRM is qualitatively similar to illusory line motion (ILM). ILM occurs when a bar is presented shortly after an initial stimulus such that the bar appears to move continuously away from the initial stimulus. IRM occurs when a second bar of a different color is presented at the same location as the first bar within a certain delay after ILM, making this second bar appear to move in the opposite direction relative to the preceding direction of ILM. Three plausible accounts of IRM are considered: a shifting attentional gradient model, a motion aftereffect (MAE) model, and a heuristic model. Results imply that IRM arises because of a heuristic about how objects move in the environment: In the absence of countervailing evidence, motion trajectories are assumed to continue away from the location where an object was last seen to move. PMID- 15876448 TI - Spectrophotometric determination of ferrate (Fe(VI)) in water by ABTS. AB - A new method for the determination of low concentrations (0.03-35 microM) of the aqueous ferrate (Fe(VI)) was developed. The method is based on the reaction of Fe(VI) with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) which forms a green radical cation (ABTS(+)) that can be measured spectrophotometrically at 415 nm (ABTS method). The reaction of Fe(VI) with ABTS has a stoichiometry of 1 : 1 in excess of ABTS (73 microM). The increase in absorbance at 415 nm for ABTS*+ generation was linear with respect to Fe(VI) added (0.03-35 microM) in buffered solutions (acetate/phosphate buffer at pH = 4.3) and was (3.40+/-0.05) x 10(4) M( 1) cm(-1). The reaction of Fe(VI) with ABTS was very rapid with a half-life time below 0.01 s at pH 4.3 and 73 microM of ABTS. This enables the ABTS method to measure Fe(VI) selectively. The residual absorbance of ABTS*+ was found to be stable in several water matrices (synthetic buffer solution and natural waters) and concentrations of Fe(VI) spiked in natural waters could be determined with high accuracy. The ABTS method can also be used as a tool to determine rate constants of reactions of Fe(VI). The second-order rate constant for the reaction of phenol with Fe(VI) was determined to be 90 M(-1) s(-1) at pH 7. PMID- 15876449 TI - Transformation products of 2-benzoxazolinone (BOA) in soil. AB - Three degradation experiments were performed to examine the formation of transformation products from 2-benzoxazolinone (BOA) in different soil types and concentrations. In two experiments using BOA in low concentration (400 microgkg( 1)) only one unidentified transformation product was found, whereas in the degradation experiment in high concentration (400 mgkg(-1)) several metabolites occurred. Two of these metabolites, 2-amino-(3H)-phenoxazin-3-one (APO); and 2 acetylamino-(3H)-phenoxazin-3-one (AAPO) were synthesized to prove their identity. This is the first time that the successive formation of these types of compounds from BOA is shown in soil. A number of other APO related transformation products were detected and provisionally characterized. The formation of APO, which is a much more biologically active compound than BOA, and the concurrent formation of a number of other APO-related metabolites in soil underline the importance of performing transformation studies as part of the evaluation of the effect of allelochemicals on weeds and soil-borne diseases. PMID- 15876450 TI - The inflammatory response to vaccination is altered in the elderly. AB - To further explore whether immune function and acute phase response are altered during ageing, the response to a mild inflammatory stress (DT-Polio-Typhim vaccination) was studied in elderly and young subjects. Cytokine production (IFN gamma, TNF-alpha, IL-6, IL-10) by whole blood cultures, circulating cytokines and acute phase proteins were analysed before and 2 days after vaccination. Prior to vaccination, only IFN-gamma production was lower in the elderly than in the young subjects due to a lower mononuclear cell number. In the same time, although in the normal range, several acute phase proteins were greater in elderly than in young subjects, suggesting a low-grade inflammatory state in the elderly. After vaccination, IFN-gamma production remained lower in the elderly than in the young, supporting an altered cell-mediated immunity with advancing age. TNF-alpha production was unaffected by either ageing or vaccination. IL-6 production was stimulated by vaccination in young subjects but not significantly in the elderly. IL-10 production was inhibited by vaccination in the elderly but not in the young. Acute phase proteins were less increased in elderly than in young subjects. Taken together, these results support a general lack of inflammatory response in the elderly exposed to an immune challenge and suggest that immune deficiency may concern both Th1 and Th2 responses. However, the interpretation must respect the limitation of small subjects number. PMID- 15876451 TI - Evaluation of two Bis-GMA analogues as potential monomer diluents to improve the mechanical properties of light-cured composite resins. AB - OBJECTIVES: The aim of this study was to investigate the influence of new diluent agents, diluent ratio and filler content, on relevant mechanical properties of several novel composite resins containing Bis-GMA as resin matrices, and to compare these with the properties of composites based on TEGDMA, a conventionally used diluent. METHODS: Two Bis-GMA analogues were synthesized and 20 experimental composite resins were prepared combining three monomer mixtures (Bis-GMA/TEGDMA, Bis-GMA/CH3 Bis-GMA and Bis-GMA/CF3 Bis-GMA), at three dilution rates (85/15, 10/90, 0/100) and three levels of hybrid filler content (barium aluminosilicate glass): 0, 10 and 35%. Flexural strength (FS), modulus of elasticity (ME) and microhardness (VHN) of the composites were evaluated. Five specimens of each material were prepared for each mechanical test, light-cured over 120 s and stored in water at 37 degrees C for 1 week. Three-point bending test was used for FS measurement and VHN was quantified by using a Vickers microindentor. Data were analyzed by ANOVA and Student-Newman-Keuls tests (P<0.05). RESULTS: Materials with CH3 Bis-GMA showed an enhanced VHN. Mean FS was higher for matrices containing TEGDMA. Overall, dilution favored FS and VHN but not ME. Filler loading specially improved ME and VHN. SIGNIFICANCE: Results correlate with an increase in the extent of polymerization due to the higher flexibility of the less viscous comonomer starting system and the hydrophobic character of the Bis GMA analogues. PMID- 15876452 TI - Effects of silicon coating on bond strength of two different titanium ceramic to titanium. AB - OBJECTIVES: This study investigated the effect of silicon coating (SiO2) by magnetron sputtering on bond strength of two different titanium ceramics to titanium. METHODS: Sixty cast titanium specimens were prepared following the protocol ISO 9693. Titanium specimens were divided into two test and control groups with 15 specimens in each. Test groups were silicon coated by the magnetron sputtering technique. Two titanium ceramics (Triceram and Duceratin) were applied on both test (coated) and control (uncoated) metal specimens. The titanium-ceramic specimens were subjected to a three point flexural test. The groups were compared for their bond strength. SEM and SEM/EDS analyses were performed on the delaminated titanium surfaces to ascertain bond failure. RESULTS: The mean bond strength of Ti-Duceratin, Ti-Triceram, Si-coated Ti Duceratin and Si-coated Ti-Triceram were 17.22+/-2.43, 23.31+/-3.18, 23.21+/-3.81 and 24.91+/-3.70 MPa, respectively. While the improvement in bond strength was 30% for Duceratin, it was statistically insignificant for Triceram. An adhesive mode of failure was observed in the Duceratin control group. In the silicoated Duceratin specimen, the bonded ceramic boundaries were wider but less than in the silicoated Triceram specimen. In the coated Triceram specimen, the ceramic retained areas were frequent and the failure mode was generally cohesive. SIGNIFICANCE: Silicon coating was significantly effective in both preventing titanium oxide layer formation and in improving bond strength for Duceratin. However, it was of less value for Triceram. PMID- 15876453 TI - A triclosan-containing compomer reduces Lactobacillus spp. predominant in advanced carious lesions. AB - OBJECTIVES: To assess the antimicrobial effect of acid etching or a triclosan containing compomer on the cultivable microflora of incompletely excavated dentinal carious lesions. METHODS: Thirty detinal lesions were opened with a diamond burr. Subsequent to removal of the softened biomass a sample of dentin was taken from the cavity floor with a round bur. Ten cavities each were treated with 36% phosphoric acid (PH) for 15s, covered with a triclosan-containing compomer (TC) or received no treatment as control (CO). All lesions were restored with a compomer composite. Sampling was performed directly after etching in the PH group and at re-entry after 6 weeks in all groups. Aliquots were plated on blood agar and selective media for Lactobacilli (Rogosa) and mutans Streptococci (MSB). Cultures were incubated anaerobically for 7 days at 37 degrees C prior to quantitative assessment and biotyping of the isolates. RESULTS: Application of phosphoric acid resulted in initial reduction of the totally cultivable microflora (p=0.006). Evaluation of the total number of cultivable microflora after 6 weeks revealed no differences between the groups (p>0.05). Lactobacilli counts were significantly lower in the TC group compared to the PH and CO groups (p<0.05). No difference was detected between the PH and CO groups after 6 weeks. SIGNIFICANCE: Phosphoric acid initially reduces the number of microorganisms in carious dentin but not in the longer term. The experimental triclosan composite suppresses Lactobacilli species over a period of 6 weeks. PMID- 15876454 TI - CML patient with rare b 2 a 3 (e 13 a 3) variant of BCR-ABL transcript: complete molecular response to imatinib. PMID- 15876455 TI - Dominant-submissive behavior as models of mania and depression. AB - This review examines the ways in which dominant-subordinate behavior in animals, as determined in laboratory studies, can be used to model depression and mania in humans. Affective disorders are mood illnesses with two opposite poles, melancholia (depression) and mania that are expressed to different degrees in affected individuals. Dominance and submissiveness are also two contrasting behavioral poles distributed as a continuum along an axis with less or more dominant or submissive animals. The premise of this article is that important elements of both mania and depression can be modeled in rats and mice based on observation of dominant and submissive behavior exhibited under well defined conditions. Studies from our own research, where dominance and submissiveness are defined in a competition test and measured as the relative success of two food restricted rats to gain access to a feeder, have yielded a paradigm that we call the Dominant Submissive Relationship (DSR). This paradigm results in two models sensitive to drugs used to treat mood disorders. Specifically, drugs used to treat mania inhibit the dominant behavior of rats gaining access to food at the expense of an opponent (Reduction of Dominant Behavior Model or RDBM), whereas antidepressants counteract the behavior of rats losing such encounters; Reduction of Submissive Behavior Model (RSBM). The validation of these models, as well as their advantages and limitations, are discussed and compared with other animal paradigms that utilize animal social behavior to model human mood disturbances. PMID- 15876456 TI - Novelty reward as a measure of anhedonia. AB - A decrease in sensitivity to pleasurable stimuli, anhedonia, is a major symptom of depression in humans. Several animal models have been developed to simulate this symptom (e.g. drug withdrawal, learned helplessness) using reward-sensitive procedures such as intracranial self-stimulation and progressive ratio responding as a measure of reward function. Recently, we introduced the use of another procedure, novel-object place conditioning in rats, to measure reward function in an associative learning situation. Withdrawal from chronic nicotine blocked a place preference conditioned by access to novel objects. This blockade was not due to impairment of object interaction, general activity, novelty detection, environmental familiarization, or expression of learning. Consequently, nicotine withdrawal directly reduced the rewarding properties of novelty. It is proposed that the novel-object place conditioning procedure could be usefully extended to other experimental situations and to genetically altered mice, so as to better understand the processes underlying changes in reward function. PMID- 15876457 TI - Immune response to and tissue localization of the Wolbachia surface protein (WSP) in dogs with natural heartworm (Dirofilaria immitis) infection. AB - Human and animal parasitic filarial nematodes, including the agent of canine and feline heartworm disease Dirofilaria immitis, harbour intracellular bacteria of the genus Wolbachia (Rickettsiaies). It is thought that these bacteria play an important role in the pathogenesis and immune response to filarial infection. Immunoglobulin G (total IgG, IgG1, IgG2) production against and immunohistochemical staining of tissues for the Wolbachia surface protein (WSP) from dogs with natural heartworm infection were evaluated. All infected dogs had significant total anti-WSP IgG levels compared to healthy controls. Interestingly, WSP was recognized by the IgG2 subclass in both microfilariemic dogs and in dogs with no circulating microfilariae (occult infection). However, microfilariemic dogs also produced gG1 antibodies. Positive staining for WSP was observed in lungs, liver and kidneys, in particular in glomerular capillaries of naturally infected dogs who had died from heartworm disease. Our results show for the first time that Wolbachia is recognized specifically by D. immitis--infected dogs and that the bacteria is released into host tissue. Furthermore, microfilariemic status appears to effect immune responses to this endosymbiont. PMID- 15876458 TI - Adaptive and innate immune responses in celiac disease. AB - Celiac disease (CD) is a complex small intestinal disorder due to a dysregulated immune response to wheat gliadin and related proteins which leads to a small intestinal enteropathy. It is generally accepted that CD is a T-cell mediated disease, in which, gliadin derived peptides, either in native form or deamidated by tissue transglutaminase, activate lamina propria infiltrating T lymphocytes which release proinflammatory cytokines. Recent studies indicate that gliadin contains also peptides able to activate an innate immune response. In particular, they induce a selective expansion of IEL, particularly TCRgamma/delta+ and CD8+TCR alpha/beta+ lymphocytes bearing the CD94 NK receptor, as well as a strong epithelial expression of MICA molecules which interact with NKG2D receptor expressed on TCRgamma/delta+ and NK cells. Most of the events of innate immune activation events are inhibited by antibodies neutralizing IL-15, thus confirming the key role of this cytokine as a mediator of intestinal mucosa damage induced by ingestion of gliadin. It remains to be established to what extent the ability of gliadin peptides to activate innate immunity relates to other biological properties exerted not only on celiac cells and tissues; the specificity of celiac patients is probably related to their genetic make up. PMID- 15876459 TI - Laryngeal inflammatory pseudotumour: an unusual cause of airway obstruction. AB - We report a rare case of a laryngeal pseudotumour in a child presenting with stridor, which was treated by endoscopic laryngeal resection, without the need for a tracheostomy. A short review of the literature is also presented. Although rare, laryngeal pseudotumour should be considered as part of the differential diagnosis of stridor and upper airway obstruction in children. PMID- 15876460 TI - Cognitive decline, neuromotor and behavioural disturbances in a mouse model for fragile-X-associated tremor/ataxia syndrome (FXTAS). AB - Carriers of premutation alleles (55-200 CGG repeats) of the fragile X mental retardation 1 (FMR1) gene are spared the major neurodevelopmental symptomatology of fragile X syndrome patients carrying a full mutation (>200 repeats). In a proportion of premutation carriers, the repeat expansion is associated with a specific neurological profile involving intention tremor, ataxia, intellectual decline compatible with dementia syndrome, Parkinsonism and autonomic dysfunction at older age, commonly referred to as fragile-X-associated tremor/ataxia syndrome (FXTAS). Typical CNS changes include hyperintense signals on T2 weighted magnetic resonance images and the presence of ubiquitin-positive intranuclear neuronal inclusions. A knock-in mouse model with a (CGG)98 repeat in the premutation range has been generated and shown to exhibit elevated Fmr1 mRNA levels and ubiquitin positive intranuclear neuronal inclusions, suggesting it may be a valid model for the human disease. Given the specific clinical profile of FXTAS patients, the expanded CGG repeat model was assessed for cognitive, behavioural and neuromotor performance at different ages (20, 52 and 72 weeks). The Morris water maze task exposed age-dependent decline of visual-spatial memory. Open field recordings revealed decreased exploration of the centre of the arena in the oldest group of expanded CGG repeat mice, potentially reflecting increased anxiety. Neuromotor tasks primarily showed decline of performance on the accelerating rotarod with age in the premutation carriers but not in control littermates. The age-dependent cognitive decline and neuromotor disturbances may be related to the progressive cognitive and behavioural difficulties observed in FXTAS patients. PMID- 15876461 TI - Tissue distribution, subcellular localization and endocrine disruption patterns induced by Cr and Mn in the crab Ucides cordatus. AB - The essential trace elements Cr and Mn are toxic at high concentrations and information about low concentration is insufficient in the literature. In polluted mangroves, the crab Ucides cordatus can represent a useful tool to assess information on the potential impact of trace elements like Cr and Mn on the environment, since this species is comestible and thus, commercially negotiated. Therefore, U. cordatus crabs were exposed in vivo to different concentrations of Cr and Mn solved in seawater and had their tissue distribution and subcellular deposits evaluated. The gill, hepatopancreas and muscle concentrations were determined by atomic absorption spectroscopy and the results showed that Cr and Mn presented the highest values in the gills rather than in the hepatopancreas and muscular tissue. Electron microscopy and analytical X-ray microanalysis revealed Cr precipitates on the gill surface, co-localized with epiphyte bacteria. In addition, since Cr and Mn did not equally accumulate in most of the tissues studied, glycemic rate of animals, which received injections of extracts of eyestalks of the contaminated crabs, were measured in order to evaluate whether the studied concentrations of Cr and Mn could produce any metabolic alteration. The results indicated that extracts of the eyestalks of crabs submitted to Cr and Mn salts and injected into normal crabs markedly influenced crustacean hyperglycemic hormone synthesis and/or release. The results are discussed with respect to sensitivity of the employed methods and the possible significance of the concentrations of Cr and Mn in the organisms. PMID- 15876462 TI - Transcriptional analysis of in vivo Plasmodium yoelii liver stage gene expression. AB - The transcriptional repertoire of the in vivo liver stage of Plasmodium has remained largely unidentified and seemingly not amenable to traditional molecular analysis because of the small number of parasites and large number of uninfected hepatocytes. We have overcome this obstruction by utilizing laser capture microdissection to provide a high quality source of parasite mRNA for the construction of a liver stage cDNA library. Sequencing and annotation of this library demonstrated expression of 623 different Plasmodium yoelii genes during development in the hepatocyte. Of these genes, 25% appear to be unique to the liver stage. This is the first comprehensive analysis of in vivo gene expression undertaken for the liver stage of P. yoelii, and provides insights into the differential expression of P. yoelii genes during this critical stage of development. PMID- 15876463 TI - Regulation of genes encoding the major surface protease of Leishmania chagasi via mRNA stability. AB - The intercoding regions between many Leishmania sp. genes regulate their mRNA expression. The MSPL mRNA, encoding a subclass of the major surface protease (MSP) of Leishmania chagasi, increases in abundance, when protein synthesis is arrested, while alpha-tubulin (alpha-TUB) mRNA and most other mRNAs do not. We found that the intercoding region between MSPL-coding regions, when cloned downstream of the beta-galactosidase reporter gene (beta-GAL), caused beta-GAL mRNA to increase 8- to 10-fold after inhibiting protein synthesis with cycloheximide. Stable L. chagasi transfectants containing hybrid MSPL/alpha-TUB intercoding regions cloned downstream of beta-GAL were made. The alpha-TUB intercoding region induced high-level baseline beta-GAL mRNA that increased only 1.3-fold after incubation with cycloheximide. In contrast, the MSPL intercoding region, as well as constructs containing nucleotides 303-505 from the MSPL 3'UTR, caused steady-state beta-GAL mRNA levels in the absence of cycloheximide that were approximately 10% of alpha-TUB constructs. These levels increased between 4.4- and 13.2-fold after cycloheximide was added. Constructs containing half of this region (303-394 or 395-505) produced intermediate levels of beta-GAL mRNA and intermediate levels of cycloheximide induction. The kinetics of cycloheximide induction of beta-GAL mRNA was similar with region 303-505 constructs as with constructs bearing the entire endogenous MSPL intercoding region. Furthermore, region 303-505 increased reporter mRNA abundance after cycloheximide by increasing mRNA half-life. Hence, we have identified a 202-nucleotide region within the MSPL 3'UTR that is in part responsible for cycloheximide induction. We hypothesize that this region may interact with labile regulatory protein factor(s). PMID- 15876464 TI - Dosimetric and clinical results of three-dimensional conformal radiotherapy for locally recurrent nasopharyngeal carcinoma. AB - PURPOSE: To assess the dosimetric and clinical results of three-dimensional conformal radiotherapy (3D CRT) for locally recurrent nasopharyngeal carcinoma (NPC). METHODS: A total of 86 patients with locally recurrent NPC were retreated with 3D CRT. The median prescribed dose was 68 Gy with 2 Gy per fractionation. Dosimetric quality was evaluated with dose distribution in planning target volume (PTV) and specified organs at risk (OAR), dose conformity index (CI) and dose homogeneity index (HI). The actuarial rate of local failure-free (LFF), overall survival (OS) and major late toxicities (MLT) were estimated with Kaplan-Meier method. Multivariate analysis for prognosis was performed using the Cox regression proportional hazards model. RESULTS: The mean dose to PTV averaged 66.8 Gy, and the dose to specified OAR was acceptable. The average value of CI and HI was 0.59 and 9.1%. The 5-year actuarial rate of LFF and OS was 71 and 40%, respectively. The 5-year actuarial incidence of MLT>or=Grade 3 and >or=Grade 4 were 100 and 49%, respectively. The major prognostic factors were T stage and the size of gross tumor volume (GTV). Advanced T stage and large GTV volume were associated with poor LFF and OS and high risk of MLT. CONCLUSION: The dosimetric quality of 3D CRT for locally recurrent NPC is generally excellent. A relatively high local control was achieved with this technique. However, the incidence of late toxicities were not found to decrease as originally expected. Early diagnosis of the recurrence and reasonable definition of the target volume are crucial to achieve a better outcome. PMID- 15876465 TI - Support for relationship between serum cholinesterase and post-traumatic stress disorder; 5-year follow-ups of victims of the Tokyo subway sarin poisoning. PMID- 15876466 TI - Efficient lactic acid production from high salt containing dairy by-products by Lactobacillus salivarius ssp. salicinius with pre-treatment by proteolytic microorganisms. AB - Lactic acid bacteria have an inefficient proteolytic system. Therefore, cultivation media which may have high protein content are usually supplemented with yeast extract or protein lysates (peptones). These additives might be conveniently replaced by in situ treatment of the cultivation medium with proteolytic enzymes or proteolytic microbes. Lactobacillus salivarius ssp. salicinius, a lactic acid bacterium species that can grow at high salt concentration, was used to ferment lactic acid in cheese whey (with 3 gl(-1) whey protein content) and lactose mother liquor (90 gl(-1) lactose, 9 gl(-1) proteins, 30 gl(-1) minerals). The contribution of protease enzymes or proteolytic microbes to acid production by lactobacilli was examined. Efficient conversion of lactose to lactic acid was obtained in the presence of additional proteolytic activity. Fastest acid production was obtained with the addition of protease enzymes. However, almost equally efficient acid production was obtained by treating the medium with Bacillus megaterium. The results show that fast and complete conversion of lactose to lactic acid can be obtained in dairy by-products without expensive additives. PMID- 15876467 TI - Combined treatment with pegylated interferon (alpha-2b) and ribavirin in the acute phase of hepatitis C virus recurrence after liver transplantation. AB - BACKGROUND/AIMS: The efficacy and safety of treatment with pegylated interferon alpha-2b (Peg-Intron, 1.5 microg/kg) and ribavirin (400-800 mg) in the acute phase of recurrent HCV after LT is presented. METHODS: Twenty-four patients (17 men) transplanted for HCV-associated cirrhosis (genotype 1b) were treated for at least 6 months and compared with 24 consecutive transplant patients (16 men) without antiviral therapy (controls). RESULTS: At completion of treatment, 14/24 treated patients (58%) achieved HCV-RNA negativity, compared to none of controls (P<0.0001). Sustained virological response (SVR) occurred in 8/23 treated patients (34.7%) who reached week 24 after treatment and none of controls (P<0.005). At 12 weeks after treatment, 15/24 patients (62.5%) had an early virological response (EVR) (seven tested HCV-RNA negative). SVR was associated with absence of corticosteroid bolus administration (P=0.01), presence of EVR (P=0.002) and absence of cytomegalovirus infection (P=0.001). Haematological adverse effects included anaemia, 17/24 cases (71%) and leukopenia, 23/24 cases (96%). One patient presented mild acute rejection that resolved by adjusting immunosuppressive dose. CONCLUSIONS: Treatment with pegylated interferon alpha-2b plus ribavirin in the acute phase of HCV reinfection yielded an EVR of 62.5% and a SVR of 34.7%. The combination was safe, with a low rate of therapy withdrawal. PMID- 15876468 TI - Hepatic steatosis in chronic hepatitis B and C: predictors, distribution and effect on fibrosis. AB - BACKGROUND/AIMS: Chronic hepatitis B (CHB) and C (CHC) are commonly associated with hepatic steatosis. The aims of this study were to investigate predictors of hepatic steatosis, and their impact on inflammation and fibrosis in CHB and CHC. METHODS: Consecutive patients with either CHB or CHC who underwent a liver biopsy at The Alfred Hospital between April and September 2002 were included. Histological analysis of liver biopsies was performed by two hepatopathologists blinded to the clinical data. RESULTS: Ninety-one patients were analysed including 17 patients with CHB and 74 with CHC. CHC genotype 3, C-peptide, glucose and waist circumference were independent predictors of extent of Brunt steatosis grade, while CHC genotype 3, C-peptide and waist circumference were independent predictors of microvesicular steatosis grade. Alcohol intake and age were predictors of hepatic fibrosis. There was a trend toward a correlation between both Brunt steatosis and microvesicular steatosis grades and fibrosis progression rate in CHC genotype non-3. CONCLUSIONS: Hepatic steatosis is common in chronic hepatitis B and C, and is associated with waist circumference, glucose, C-peptide and chronic hepatitis C genotype 3. Steatosis grade appears to relate to hepatic fibrosis progression rate in chronic hepatitis C genotype non 3. PMID- 15876469 TI - The RL-ET-14 cell line mediates expression of glutamine synthetase through the upstream enhancer/promoter region. AB - BACKGROUND/AIMS: The expression of glutamine synthetase (GS) in the mammalian liver is confined to the hepatocytes surrounding the central vein and can be induced in cultures of periportal hepatocytes by co-cultivation with the rat liver epithelial cell line RL-ET-14. We exploited these observations to identify the regulatory regions of the GS gene and the responsible signal-transduction pathway that mediates this effect. METHODS: Fetal hepatocytes of wild-type or GS transgenic mice were co-cultured with RL-ET-14 cells to induce GS expression. Small-interfering RNA was employed to silence beta-catenin expression in the fetal hepatocytes prior to co-culture. RESULTS: Co-cultivation of RL-ET-14 cells with fetal mouse hepatocytes induced GS expression 4.2-fold. The expression of another pericentral enzyme, ornithine aminotransferase and a periportal enzyme, carbamoylphosphate synthetase, were not affected. Co-culture of RL-ET-14 cells with transgenic fetal mouse hepatocytes demonstrated that GS expression was induced via its upstream enhancer located at -2.5 kb and that the signal mediator required a functional beta-catenin pathway. CONCLUSIONS: The 'RL-ET-14' factor specifically induces GS expression, working via its upstream enhancer in a beta catenin-dependent fashion. PMID- 15876470 TI - Applicability of adult-to-adult living donor liver transplantation. AB - BACKGROUND/AIMS: The applicability of adult-to-adult living donor liver transplantation has not been established yet. We report the first data in a European center of the process leading to this procedure from the first moment the patients were informed about it. METHODS: In phase 1 of the process, 121 adult patients enlisted for cadaveric liver transplantation and their relatives were informed of the technical aspects, advantages and risks of living donor liver transplantation, and the essential criteria for living donation. In phase 2, potential donors identified in phase 1 were evaluated in depth. RESULTS: Twenty-one (17%) patients underwent living donor liver transplantation. This procedure was not performed in 60 patients (50%) for reasons concerning the patients themselves, especially their refusal to receive living donor liver transplantation from a relative (30%). Forty patients (33%) did not undergo living donor liver transplantation for reasons concerning potential donors: donors were not identified (14%), declined the donation (13%), or were refused for technical reasons (6%). The expected waiting time to transplantation was longer in patients who underwent living donor liver transplantation than in those who did not. CONCLUSIONS: The applicability of adult-to-adult living donor liver transplantation is low, mainly because of reasons related to potential recipients. PMID- 15876471 TI - Factors affecting the uptake of screening: a randomised controlled non inferiority trial comparing a genotypic and a phenotypic strategy for screening for haemochromatosis. AB - BACKGROUND/AIMS: Haemochromatosis provides an example where a novel pragmatic genotypic screening strategy may be compared with a phenotypic strategy assessing factors affecting uptake, feasibility and cost. METHODS: A randomised controlled 'non-inferiority' trial testing the hypothesis that the uptake of testing in the genotypic strategy would not be inferior to the uptake in a phenotypic screening strategy. Three thousand individuals aged 30-70 were randomly selected and randomly allocated (stratified by age and sex) to one of two screening strategies. Phenotypic-transferrin saturation on blood sample taken at GP surgery or genotypic-saliva sample taken at home; followed in screen positive individuals with assessment of iron status and genotyping. RESULTS: The difference in uptake between the two strategies was 3.4% (95% CI=0.5-6.8). Uptake was low (32%) and least in young men from socially deprived areas. Phenotypic screening was least costly. CONCLUSIONS: In this study, investigating the uptake of screening for a treatable disease in primary care, the uptake of screening with the genotypic strategy was not inferior to that in the phenotypic strategy. The poor uptake in younger men would further limit the effectiveness of screening for haemochromatosis and may have implications for other screening programmes targeted to this group. PMID- 15876472 TI - Cue-elicited craving for food: a fresh approach to the study of binge eating. AB - Recent research has indicated that craving for food can be elicited by exposure to food cues, suggesting that exposure to food cues may represent a useful experimental paradigm to investigate mechanisms related to binge eating. The first objective of the present research was to replicate previous reports that exposure to food cues elicits craving for food. In addition, this investigation was designed to extend the extant literature by testing the effects of 'priming' portions of food, by examining the association between reactivity to food cues and indicators of binge eating, and to examine the role of a putative genetic factor previously found to be associated with cue-elicited craving for alcohol and tobacco. In Study 1, 48 individuals completed measures of craving and mood after exposure to control cues, after exposure to food cues, and after consuming each of three small portions of food. In Study 2, 31 individuals with subclinical symptoms of binge eating completed the same procedures. The results suggested that food cues reliably elicited craving, increased attention to the cues, and decreased positive affect in both samples, although reactivity was greater among the sample with greater eating pathology. Correlational analyses suggested that reactivity to food cues was correlated with binge eating and body mass index among women but not men. Results also suggest that the DRD4 VNTR polymorphism influences cue-elicited craving for food, although the influence of the DRD4 may depend on the population under study. PMID- 15876473 TI - Stronger anti-HIV-1 activity of C-peptide derived from HIV-1 89.6 gp41 C-terminal heptad repeated sequence. AB - C34-LAI containing amino acids 118 to 151 of the HIV-1(LAI) gp41 ectodomain exhibits potent anti-HIV-1 activity. However, the N-terminal halves of C34 peptides vary more according to the HIV-1 strain than the C-terminal halves. Therefore, an analysis was conducted on the anti-HIV-1 activities of the C34 peptides derived from various HIV-1 strains. C34-89.6 exhibited the strongest anti-HIV-1 activity among the C34 peptides tested. Interestingly, its N-terminal half was more acidic than those of the other C34 peptides, whereas its C-terminal half was more basic. Since the C-peptides derived from the HIV-1(LAI) strain are used extensively, the anti-HIV-1 activities of these peptides were compared between the HIV-1 strains 89.6 and LAI. When using chimeric peptides, it was found that the C-terminal basic region of C34-89.6 was more critical than its N terminal basic region. The anti-HIV-1 activity of T20-89.6 and C28-89.6 was also stronger than that of T20-LAI and C28-LAI, respectively. The anti-HIV-1 activity of C28-89.6 was weakened when the C-terminal basic residues were changed to the corresponding residues of C28-LAI. However, no conformational differences were found among the C28 peptides. Accordingly, these results imply that introducing the C-terminal basic residues of the HIV-1 89.6 C-peptide may be useful for developing potent anti-HIV-1 drugs. PMID- 15876474 TI - Angiotensin II induced increase in frequency of cytosolic and nuclear calcium waves of heart cells via activation of AT1 and AT2 receptors. AB - The aim of this work is to verify if Angiotensin II (Ang II) affects the frequency of spontaneous cytosolic and nuclear Ca2+ waves in chick embryonic cardiomyocytes and if this effect is mediated via the activation of AT1 and/or AT2 receptors. Using the rapid scan technique of confocal microscopy, we observed that Ang II (10(-8)M) increases the frequency of cytosolic and nuclear Ca2+ waves. This effect was accompanied by a decrease in the amplitude of nuclear Ca2+ waves and an absence of effect on the amplitude of cytosolic Ca2+ waves. The effect of the octapeptide on both frequency and amplitude of the nuclear waves was prevented by the AT1 receptor antagonist L158809. However, blockade of the AT2 receptor using the antagonist PD123319 (10(-7)M) only prevented the effect of Ang II on the frequency of Ca2+ waves. Furthermore, the effect was prevented by both a PKC inhibitor (bisindolylmaleimide) and a PKC activator (phorbol 12,13 dibutyrate). In addition, the Ang II effect was not prevented by the blocker of the pacemaker current If. These results demonstrate that Ang II, via the activation of its receptors AT1 and AT2, affects the frequency of spontaneous Ca2+ waves and this effect seems to be mediated by the PKC pathway. PMID- 15876475 TI - Design of novel melanotropin agonists and antagonists with high potency and selectivity for human melanocortin receptors. AB - alpha-MSH and gamma-MSH are the natural endogenous hormones for the human melanocortin-1, 3, 4 and 5 receptors (hMC1R, hMC3R, hMC4R and hMC5R). These and more potent, stable and prolonged acting analogues such as NDP-alpha-MSH, MT-II and SHU-9119 are not very receptor selective. To develop potent and selective agonist and antagonist ligands for the melanocortin receptors we have used state of-the-art biophysical studies, computational chemistry, and design of conformational and topographical constraints with novel templates. PMID- 15876476 TI - Isatin, an endogenous monoamine oxidase inhibitor, triggers a dose- and time dependent switch from apoptosis to necrosis in human neuroblastoma cells. AB - Isatin is an endogenous indole that is increased in stress, inhibits monoamine oxidase (MAO) B and improves bradykinesia and striatal dopamine levels in rat models of Parkinson's disease. Consequently, it has been suggested that isatin might be a possible treatment for Parkinson's disease although little is known about its effects on neural cell growth and survival. The aim of this study was to investigate the survival of dopaminergic human neuroblastoma (SH-SY5Y) cells following treatment with increasing concentrations of isatin. SH-SY5Y cells were exposed to isatin for defined time points, after which cell survival was determined by MTT assay. A combination of Annexin V binding and propidium iodide (PI) exclusion was used to distinguish apoptosis from necrosis in flow cytometry experiments and FACS profiles of permeabilised PI-labelled cells were employed for the assessment of cell cycle distribution. Isatin treatment (1-400 microM) for 24h induced a significant dose-dependent increase in MTT metabolism by SH SY5Y cells in culture, but this was not due to an increase in cell division. At the higher concentrations (200-400 microm) isatin triggered cell death, although MTT metabolism was still increased in the culture, suggesting that surviving cells were hypermetabolic. Following a longer (48 h) exposure, isatin was found to cause cell death in a dose-dependent manner; at lower concentrations (50 microM), the predominant mode of cell death was apoptosis while at the highest concentration (400 microm) increasing numbers of necrotic cells were also evident. Thus, in dopaminergic SH-SY5Y cells isatin induces cell death in dose- and time-dependent manner. This death occurred as a continuum of survival, apoptosis and necrosis. Our results re-emphasise that caution should be exercised when considering high doses of isatin as a putative anti-Parkinson's disease therapeutic. PMID- 15876477 TI - Valinomycin-induced apoptosis of human NK cells is predominantly caspase independent. AB - Human NK cells are sensitive to the exogenous toxic compound valinomycin. This toxin, produced by Streptomyces griseus in moisture damaged buildings, induces apoptosis by dissipating the membrane potential in mitochondria. In this paper, we show that valinomycin-induced apoptosis involves two different pathways in human NK cells: the predominant one is caspase-3 independent and the other caspase-3 dependent. Resting human NK cells were found to contain high amounts of active caspase-3 as compared to the T cells in which high caspase-3 activity has been shown only after stimulation. Exposure to valinomycin did not alter the caspase-3 activity of human NK cells but induced nucleosomal fragmentation of DNA. General caspase inhibitor, Z-VAD-FMK, inhibited completely the caspase-3 activity, reduced DNA cleavage but did not prevent the spontaneous or valinomycin induced apoptosis of NK cells. The endogenous high caspase-3 had only a slight effect on the major functions of human NK cells, i.e. cytotoxicity or gamma-IFN production, giving us a reason to suspect that the biological role of caspase-3 in NK cells could be the elimination of potentially harmful NK clones through apoptosis. PMID- 15876478 TI - The role of DNA response elements as allosteric modulators of steroid receptor function. AB - Steroid receptors are ligand-activated transcription factors which control the expression of their target genes by binding to specific DNA elements. Consensus response elements have been delineated for the glucocorticoid, androgen, progesterone and mineralocorticoid receptors on one hand (steroid response element, SRE) and for the estrogen receptor on the other hand (estrogen response element, ERE). Small variations in these sequences not only affect the binding but may also have a dramatic impact on the transcriptional activity of steroid receptors. It has now become obvious that DNA response elements do not merely tether regulatory proteins to control regions of target genes but may additionally impart conformational changes onto the DNA-binding domain as well as to neighbouring domains of steroid receptors. This in turn will create unique platforms for selective recruitment of cofactors and possibly for induction of modifications in local chromatin architecture. An additional level of complexity is added by the frequent presence of multiple response elements in gene promoter regions. The allosteric effects of DNA response elements on steroid receptors may be essential for differential gene expression and this offers interesting perspectives for the identification of selective modulators. PMID- 15876479 TI - Cloning, characterization and expression of the D2 dopamine receptor from the tilapia pituitary. AB - A full-length cDNA encoding a dopamine receptor (DA-R) was obtained from the pituitary of tilapia (ta). This cDNA encodes a protein of 469 amino acids that exhibits the typical arrangement of GPCR. The taDA-R shows high similarity to the DA-Rs of mullet and fugu, and over 70% similarity to Xenopus, mouse and turkey D2 DA-Rs. Northern blot analysis revealed transcript for a single transcript in the pituitary, of approximately 3 kb. In a Southern analysis, the tilapia probe recognized specific bands in the genomic DNA of both mullet and catfish, suggesting high similarity between the corresponding genes. Phylogenetic analysis clearly aligned the taDA-D2-R with all vertebrate D2-like receptor sequences cloned to date, and it was therefore designated taDA-D2-R. taDA-D2-R was transiently expressed in COS-7 cells together with the reporter construct CRE luciferase. Addition of the specific D2 dopamine agonists quinpirole or bromocriptine, in the presence of forskolin, led to a dose-dependent decrease in forskolin-induced cAMP levels. Both agonists yielded the same maximal inhibition (around 40%). However, the potency of taDA-D2-R for bromocriptine was higher than for quinpirole. As established for mammalian D2-like receptors, stimulation of the taDA-D2-R with quinpirole triggers pertussis-toxin-sensitive Gi/o-mediated, but not Gs-mediated signaling. In contrast to mammals, PCR analysis gave no evidence of alternative splicing in taDA-D2-R. Pharmacological and genetic manipulation of the taDA-D2-R should enable us to better define its physiological role and to further explore the usefulness of fish as a model system for understanding dopaminergic function in higher organisms. PMID- 15876480 TI - BRCA1-2 mutations in breast cancer: identification of nine new variants of BRCA1 2 genes in a population from central Western Spain. AB - We carried out a mutational analysis of BRCA1 and BRCA2 genes in 103 individuals from a population in Western Central Spain and we identified nine new variants: two truncating mutations in BRCA2 [2604C>A (Y792X), 8873del4], three missense mutations in BRCA2 [677A>G (H150R), 958G>A (D224N) and 3398A>G (K1057R], and four silent mutations, two in BRCA1 [1115T>G (R332R) and IVS24+36 C>G], and two in BRCA2 [2583T>A (I785I) and 7854G>A (T2542T)]. In two unrelated families of our population, we identified the BRCA1 1806C>T (Q563X) mutation, which is considered to be a Swedish founder mutation. BRCA1 1806C>T (Q563X) and BRCA2 3036del4 gene mutations were the most frequent in our series. PMID- 15876481 TI - Mutation and expression analysis of LZTS1 in ovarian cancer. AB - LZTS1 has been shown to have tumour suppressor activities against prostate and breast cancer and is located within a region of frequent loss of heterozygosity (LOH) at 8p22 in ovarian cancer. We have analysed the expression of LZTS1 in ovarian cancer and found no evidence of loss of expression relative to normal ovarian surface epithelial cells. We have also analysed the coding region of the LZTS1 gene in 87 primary ovarian adenocarcinomas by DHPLC and detected a single silent somatic mutation. These data indicate that LZTS1 is not the target of LOH at 8p22 in ovarian cancer. PMID- 15876482 TI - A new medium-term rat colon bioassay applying neoplastic lesions as endpoints for detection of carcinogenesis modifiers-validation with known modifiers. AB - We have established a medium-term colorectal carcinogenesis rat model initiated with 1,2-dimethylhydrazine (DMH) followed by dextran sodium sulfate (DSS) treatment, featuring induction of neoplastic lesions within 10 weeks. In the present study, we examined its ability to detect modification of colon lesion development with 10- or 20-week experimental periods. F344 male rats were given three subcutaneous injections of DMH (40 mg/kg b.w.) in a week followed by free access to drinking water containing 1% DSS for a week. One week after this regimen, basal diet alone, basal diet containing 0.04% nimesulide or 2% lactoferrin as known inhibitors, 0.3% deoxycholic acid (DCA) as a promoter or 1.5% 1-hydroxyanthraquinone (1-HA) as a carcinogen were supplied. At week 10, the incidence and multiplicity of combined adenomas and adenocarcinomas were significantly (P < 0.05 or 0.01) decreased by nimesulide and lactoferrin, and values for adenomas were significantly (P < 0.01) increased in the 1-HA group. There was no clear change in the DCA group. At week 20, multiplicity and volume of the tumors were significantly (P < 0.01 or 0.05) decreased by nimesulide, but no effect was now evident with lactoferrin. Multiplicity and volume of tumors were significantly (P < 0.01) increased in 1-HA group and a similar tendency was apparent (P = 0.08) with DCA. It is concluded that this system offers a useful tool for detection of colorectal carcinogenesis modifiers within 10-20 weeks, pending further studies for verification employing other model chemicals. PMID- 15876483 TI - Endostatin gene transfer in murine lung carcinoma cells induces vascular endothelial growth factor secretion resulting in up-regulation of in vivo tumorigenecity. AB - Endostatin (ED) is a carboxyl-terminal fragment of type XVIII collagen with a strong anti-angiogenic activity. The purpose of this study is to determine the effect of ED gene transfer into lung cancer cells on in vivo tumor growth in a murine model. The murine lung cancer cell line, Lewis Lung Carcinoma (LLC), was transfected with ED gene to express and secrete ED. After clones were selected to secrete ED, several stable transfectants with ED gene (LLC/ED) and control transfectants (LLC/Mock) were established. In vitro proliferation of these transfectants demonstrated similar growth speed. In contrast to previous reports, in vivo subcutaneous tumorignecity of LCC/ED transfectants was significantly greater than that of LLC/Mock transfectants. Immunohistochemical staining analysis demonstrated that ED gene transfer induced angiogenesis, suggesting coinduction of another gene implicated for neovascularization. As expected, LLC/ED transfectants secreted not only ED but also vascular endothelial growth factor (VEGF) to a much greater degree than LLC/mock transfectants. Interestingly, culture supernatants of LLC/ED cells enhanced in vitro proliferation of human umbilical vein endothelial cells (HUVEC) to a much greater degree than those of LLC/Mock cells. These results indicate that ED gene transfer in murine lung carcinoma cells induces VEGF secretion, resulting in enhancement of in vivo tumorigenecity in the murine model. More attention should be paid for ED gene therapy into lung cancer cells since it may influence other proteins secretion, which upregulates angiogenesis. PMID- 15876484 TI - A speed congenic rat strain bearing the tongue cancer susceptibility locus Tscc1 from Dark-Agouti rats. AB - We previously reported that Dark-Agouti (DA) rats are highly susceptible to 4 nitroquinoline 1-oxide (4NQO)-induced tongue cancer (TC), whereas Wistar/Furth (WF) rats are barely susceptible. Linkage analysis of reciprocal (DAxWF)F2 rats demonstrated five quantitative trait loci, Tongue squamous cell carcinoma 1-5 (Tscc1-5) determining the size and number of the TCs. The major susceptibility locus Tscc1 is mapped on rat chromosome 19. In the present study, we used a marker-assisted speed congenic procedure to construct WF.DA-Tscc1 (WF-T1D) rats, i.e. WF rats carrying a DA-derived Tscc1 chromosomal segment, and evaluated the effect of a single Tscc1 on 4NQO-induced tongue carcinogenesis. In WF-T1D rats, the incidence, number and size of 4NQO-induced TCs were significantly higher than those in WF rats, indicating that the introgressed segment contains one of the susceptibility loci for 4NQO-induced TCs from DA rats. Detection of a single nucleotide polymorphism in NQO1, one of the Tscc1 candidate genes, enabled us to map NQO1 in the Tscc1 segment between D19Wox8 and D19Wox7 on chromosome 19. Possible relevance of NQO1 polymorphism to TC susceptibility is discussed. PMID- 15876485 TI - Increased thromboxane B(2) levels are associated with lipid peroxidation and Bcl 2 expression in human lung carcinoma. AB - There is little information regarding simultaneous investigations of thromboxane A(2) (TXA(2)) lipid peroxidation and Bcl-2, three cancer-related agents, and analyses of their relationships in lung cancer. The present study was to study thromboxane B(2) (TXB(2)), a stable metabolite of TXA(2), lipid peroxidation and Bcl-2 expression in 52 non-small cell lung carcinoma (NSCLC) tissue samples. The level of thiobarbituric acid reactive substances (TBARS), an index for lipid peroxidation was significantly increased in the lung tumor tissues, compared with non-tumor tissues. TXB(2) was much higher in the tumor tissues than non-tumor tissues. Interestingly, the concentration of TXB(2) in samples from those who smoked was higher than that from those who did not smoke. The expression of Bcl-2 was significantly elevated in the tumor tissues, compared to the non-tumor tissues. There was also a positive correlation between TXB(2) and TBARS in tumor tissues; advanced stage cancers had higher levels of TXB(2). This finding supports the idea that TXB(2) may have a role in promoting tumor growth. In conclusion, our study demonstrates that the production of TXB(2) is increased in lung tumor tissues and that such an increase can result in lipid peroxidation which may be met by an elevation in Bcl-2 expression. PMID- 15876486 TI - Establishment of an apoptosis-sensitive rat mammary carcinoma cell line with a mutation in the DNA-binding region of p53. AB - Seven mammary carcinoma cell lines were established from 7,12 dimethylbenz[a]anthracene-induced tumors developed in a human c-Ha-ras transgenic rat. Without apoptotic stimuli, a large amount of p53 protein was detected in the C11 cell line (C11), whereas all cell lines expressed variable levels of the assayed death receptor/ligand, bcl-2 family and p53 cascade-related genes. The p53 gene in C11 had a mutation at codon 246, in the DNA-binding region of p53. Transcriptional activity of the mutant protein appeared to be lower than that of the wild-type p53. Despite the presence of p53 mutation, C11 was more sensitive to apoptosis triggered by etoposide, paclitaxel and staurosporine than the cell lines expressing wild-type p53. These data suggest that the apoptosis induced by intracellular injury occurs via the transcriptionally impaired mutant p53 in C11. PMID- 15876487 TI - CYP3A5 gene polymorphism and risk of prostate cancer in a Japanese population. AB - The CYP3A5 gene (CYP3A5) encodes the cytochrome P450 3A5, which catalyzes the 6beta-hydroxylation of testosterone. We explored association between the CYP3A5 A6986G polymorphism and a risk of prostate cancer in 260 prostate cancer patients, 199 BPH patients and 212 male controls. The CYP3A5 gene polymorphism did not influence significantly a risk of developing of prostate cancer in general. However, compared with males with the GG genotype, those with the AA genotype had a 0.23-fold decreased risk of developing low-grade prostate cancer (P=0.023), and a 0.31-fold decreased risk of developing localized (stages A-C) prostate cancer (P=0.044). The CYP3A5 A6986G polymorphism may be specifically associated with a decreased risk of low-grade or early stage prostate cancer. PMID- 15876488 TI - The GC/AED studies on the reactions of sulfur mustard with oxidants. AB - A gas chromatograph coupled with an atomic emission detector was used to identify and to determine the products formed on oxidation of sulfur mustard. The oxidation rate and the resulting oxidates were studied in relation to oxidant type and reaction medium parameters. Hydrogen peroxide, sodium hypochlorite, sodium perborate, potassium monopercarbonate, ammonium peroxydisulfate, potassium peroxymonosulfate (oxone), and tert-butyl peroxide were used as oxidants. Oxidations were run in aqueous media or in solvents of varying polarities. The oxidation rate was found to be strongly related to oxidant type: potassium peroxymonosulfate (oxone) and sodium hypochlorite were fast-acting oxidants; sodium perborate, hydrogen peroxide, ammonium peroxydisulfate, and sodium monopercarbonate were moderate oxidants; tert-butyl peroxide was the slowest acting oxidant. In non-aqueous solvents, the oxidation rate was strongly related to solvent polarity. The higher the solvent polarity, the faster the oxidation rate. In the acid and neutral media, the mustard oxidation rates were comparable. In the alkaline medium, oxidation was evidently slower. A suitable choice of the initial oxidant-to-mustard concentration ratio allowed to control the type of the resulting mustard oxidates. As the pH of the reaction medium was increased, the reaction of elimination of hydrogen chloride from mustard oxidates becomes more and more intensive. PMID- 15876489 TI - Pilocarpine-induced status epilepticus: monoamine level, muscarinic and dopaminergic receptors alterations in striatum of young rats. AB - Behavioural changes, muscarinic and dopaminergic receptors density and levels of monoamines were measured in striatum of rats after pilocarpine-induced status epilepticus (SE). Wistar rats at the age of 21 days were treated with pilocarpine (400mg/kg; subcutaneously) whilst the control group was treated with 0.9% saline (s.c.). Both groups were sacrificed 1h following the treatment. SE induced a muscarinic receptor downregulation of 64% in pilocarpine group. This effect was also observed to be 57% in D(1) and 32% in D(2). In the dissociation constant (K(d)) values in muscarinic and D(1) receptor no alterations were verified. On the other hand, the K(d) value for D(2) was observed to increase 41%. High performance liquid chromatography determinations showed 63, 35, 77 and 64% decreases in dopamine, 3-methoxy-phenylacetic acid, serotonin and 5 hydroxyindoleacetic acid contents, respectively. The homovanilic acid level was verified to increase 119%. The noradrenaline content was unaltered. A direct evidence of monoamine levels alterations can be verified during seizure activity and receptor density changes appear to occur in an accentuated way in immature brain during the estabilishment of SE induced by pilocarpine. PMID- 15876490 TI - Alteration of posture-related cortical potentials in mild traumatic brain injury. AB - This paper presents additional evidence showing the persistent functional deficits in concussed athletes as revealed by altered movement-related cortical potentials (MRCP) preceding whole body postural movements at least 30 days post injury. Eight student-athletes participated in this study (a) prior to injury; and (b) 3, 10 and 30 days after MTBI. EEG was recorded while subjects produced static balance tasks and dynamic postural movements. All subjects were cleared for sport participation within 10 days post-injury based upon neurological and neuropsychological assessments as well as upon clinical symptoms resolution. There was a persistent reduction of MRCP amplitude prior to initiation of postural movement up to 30 days post-injury, although abnormal postural responses basically recovered within 10 days post-injury. The frontal lobe MRCP effects were larger than posterior areas. This supports the notion that behavioral symptoms resolution may not be indicative of brain injury resolution. Overall, persistent alteration of movement-related cortical potentials after MTBI may indicate residual disturbance of neuronal networks involved in preparation and execution of postural movements and a lower threshold for brain re/injury. PMID- 15876491 TI - Acupoint-specific fMRI patterns in human brain. AB - Specific central nervous system (CNS) responses to acupuncture have recently attracted attention. It is important to understand the differences in fMRI images of the brain evoked by acupuncture at an acupoint and at a nearby "sham" point. Here, we report analyses of fMRI images of the brains of 37 healthy volunteers in response to acupuncture at Liv3 (Taichong) and LI4 (Hegu) versus their sham points. We found common activation areas in response to Liv3 or LI4 acupuncture in the middle temporal gyrus and cerebellum, along with deactivation areas in the middle frontal gyrus and inferior parietal lobule, compared with the effects of acupuncture at sham points. Acupuncture at Liv3 evoked specific activation at the postcentral gyrus, posterior cingulate, parahippocampal gyrus, BA 7, 19 and 41, but deactivation at the inferior frontal gyrus, anterior cingulate, BA 17 and 18, compared with acupuncture at its sham point. Acupuncture at LI4 evoked specific activation at the temporal pole, but deactivation at the precentral gyrus, superior temporal gyrus, pulvinar and BA 8, 9 and 45, compared with acupuncture at its sham point. These observations reveal that acupuncture at acupoints induces specific patterns of brain activity, and these patterns may relate to the therapeutic effects of acupuncture. PMID- 15876492 TI - Suppression of stress immobilization-induced phosphorylation of ERK 1/2 by biting in the rat hypothalamic paraventricular nucleus. AB - We have previously reported that acute immobilization stress induces Fos protein. Fos protein is generally used as a marker for neuronal activity and has been linked to phosphorylation of extracellular signal-regulated protein kinase 1/2 (pERK1/2), in the hypothalamic paraventricular nucleus (PVN). Biting behavior during the period of stress reduced the expression of Fos protein. The present immunohistochemical study was designed to determine whether acute immobilization stress induces pERK1/2 in the PVN, and whether the stress-induced pERK1/2 was attenuated by simultaneous biting behavior. Acute immobilization stress, in increments of up to 15min, produced detectable amounts of pERK1/2 that were proportional to the interval of stress. Biting during the acute immobilization stress significantly reduced the amount of detectable pERK1/2. These results suggest that biting activity during acute stress inhibits pERK1/2 in this region of the brain. It is feasible that the neuronal cellular response to acute stress is regulated, in some part, by inhibition of pERK1/2 by biting. PMID- 15876493 TI - Assessment of the effectiveness of peripheral administration of morphine with local articaine anaesthesia for surgery in inflamed oral and maxillofacial tissues. AB - The controversy surrounding clinical trials of peripherally applied morphine with local anaesthetic and the attendant ambiguous results led to a study of our own clinical material. The aim of the study was to assess the effectiveness of peripheral administration of morphine with local articaine anaesthesia in inflamed oral and maxillofacial tissues. Sixty patients who qualified for the randomized, double-blinded study were randomly divided into two groups. Group 'LA' received a standard local anaesthetic solution (articaine plus epinephrine) while group 'LA-Mo' received the standard solution with an addition of 1 mg of morphine. Pain intensity was assessed using the Visual Analogue Scale, before and directly after surgery as well as at 1, 2, 6, 12, 24 and 48 h after completion of surgery. Furthermore, supplemental consumption of the prescribed analgesic was recorded. Despite a very similar average level of initial pain, there was a marked difference between the groups in the pain level during surgery. Moreover, during the next 12 h, there were significant differences observed in the level of pain between both groups. There was also considerable difference between both groups in the time of first analgesic intake and the total amount of analgesic. Our results show that modified local anaesthesia may be of benefit for the relief of operative and post-operative pain and may also help reduce analgesic intake after oral surgery. PMID- 15876494 TI - Age effects on pain thresholds, temporal summation and spatial summation of heat and pressure pain. AB - Experimental data on age-related changes in pain perception have so far been contradictory. It has appeared that the type of pain induction method is critical in this context, with sensitivity to heat pain being decreased whereas sensitivity to pressure pain may be even enhanced in the elderly. Furthermore, it has been shown that temporal summation of heat pain is more pronounced in the elderly but it has remained unclear whether age differences in temporal summation are also evident when using other pain induction methods. No studies on age related changes in spatial summation of pain have so far been conducted. The aim of the present study was to provide a comprehensive survey on age-related changes in pain perception, i.e. in somatosensory thresholds (warmth, cold, vibration), pain thresholds (heat, pressure) and spatial and temporal summation of heat and pressure pain. We investigated 20 young (mean age 27.1 years) and 20 elderly (mean age 71.6 years) subjects. Our results confirmed and extended previous findings by showing that somatosensory thresholds for non-noxious stimuli increase with age whereas pressure pain thresholds decrease and heat pain thresholds show no age-related changes. Apart from an enhanced temporal summation of heat pain, pain summation was not found to be critically affected by age. The results of the present study provide evidence for stimulus-specific changes in pain perception in the elderly, with deep tissue (muscle) nociception being affected differently by age than superficial tissue (skin) nociception. Summation mechanisms contribute only moderately to age changes in pain perception. PMID- 15876495 TI - Human surrogate models of neuropathic pain. PMID- 15876496 TI - The outcome of pain related undergraduate teaching in Finnish medical faculties. AB - Little is known about how other than cancer pain related issues are represented in medical education. A standardised questionnaire was mailed to all medical students who graduated from the five Finnish medical schools in 2001. A total of 387 students received the questionnaire and 41% responded. The students had to evaluate the quantity and the quality of pain teaching. The availability and the participation in the advanced courses or research in pain medicine were asked. The students reported how the IASP curriculum on pain had been covered during the studies. Two clinical cases were presented for diagnosis and treatment. In addition to integrated pain teaching, specific pain education was received by 27% of the students. The departments of anaesthesiology were reported as the major deliverers of teaching of pain. The overall ratings of the pain-related teaching of the faculties varied from 3.4 to 4.6 on a scale of 10. Anatomy, biochemistry, physiology and pharmacology of pain were covered well. The definitions of pain, pain research, sociological issues, paediatric, geriatric and mentally retarded patients' pain were taught most poorly. Only 34% of the students had been offered advanced studies and 15% had been offered research projects in pain medicine. The lack of teaching about the concept of a multidisciplinary pain clinic was recognised by almost all students. The clinical problems were excellently solved. In conclusion, the IASP curriculum is well covered in the present programmes in the Finnish medical faculties. However, the quality and the methods of teaching still need improvement. PMID- 15876497 TI - Multislice theory of fast electron scattering incorporating atomic inner-shell ionization. AB - It is demonstrated how atomic inner-shell ionization can be incorporated into a multislice theory of fast electron scattering. The resulting theory therefore accounts for both inelastic scattering due to inner-shell ionization and dynamical elastic scattering. The theory uses a description of the ionization process based on the angular momentum representation for both the initial and final states of the atomic electron. For energy losses near threshold, only a small number of independent states of the ejected atomic electron need to be considered, reducing demands on computing time, and eliminating the need for tabulated inelastic scattering factors. The theory is used to investigate the influence of the collection aperture size on the spatial origin of the silicon K shell EELS signal generated by a STEM probe. The validity of a so-called local approximation is also considered. PMID- 15876498 TI - Anxiety disorders among patients with co-occurring bipolar and substance use disorders. AB - Bipolar and substance use disorders are known to co-occur frequently, but limited attention has been paid to anxiety disorders that may accompany this dual diagnosis. Therefore, we examined the prevalence and nature of anxiety disorders among treatment-seeking patients diagnosed with current bipolar and substance use disorders, and investigated the association between anxiety disorders and substance use. Among 90 participants diagnosed with bipolar disorder I (n = 75, 78%) or II (n = 15, 22%), 43 (48%) had a lifetime anxiety disorder, with post traumatic stress disorder (PTSD) occurring most frequently (n = 21, 23%). We found that those with PTSD, but not with the other anxiety disorders assessed, began using drugs at an earlier age and had more lifetime substance use disorders, particularly cocaine and amphetamine use disorders, than those without PTSD. Further examination revealed that (1) most participants with PTSD were women, (2) sexual abuse was the most frequently reported index trauma, and (3) the mean age of the earliest index trauma occurred before the mean age of initiation of drug use. Our findings point to the importance of further investigating the ramifications of a trauma history among those who are dually diagnosed with bipolar and substance use disorders. PMID- 15876499 TI - Faecal steroid metabolites for non-invasive assessment of reproduction in common warthogs (Phacochoerus africanus), red river hogs (Potamochoerus porcus) and babirusa (Babyrousa babyrussa). AB - The objectives of this study were to analyse faecal steroid metabolites in African and South East Asian pig species kept in European zoos. Species studied were the warthog (Phacochoerus africanus), the red river hog (Potamochoerus porcus) and the babirusa (Babyrousa babyrussa). Faecal samples were collected 1-3 times per week from non-pregnant and pregnant captive female warthogs (n = 9), red river hogs (n = 7) and babirusas (n = 5). Enzyme-immunoassays for faecal progesterone, androgen, and oestrogen metabolites, were tested for their ability to determine follicular and luteal phases. In all three species, oestrous cycles could be monitored with 20alpha-OH- and 20-oxo-pregnane assays. In contrast, oestrogens and androgens were not useful in characterising follicular activity during the oestrous cycle in any species. Faecal 20alpha-OH- and 20-oxo-pregnane values were significantly correlated. Faecal pregnane concentrations revealed species-specific differences. Luteal phase values of 20alpha-OH-pregnanes were considerably higher than 20-oxo-pregnanes; 20alpha-OH-pregnanes were in the range of 3-10 microg/g in warthogs and red river hogs, whereas concentrations were 30 200 microg/g faeces in the babirusa. Regular oestrus cycles had a length of about 35 days in all three species studied. Results indicated a seasonal influence on the occurrence of reproductive cycles in the warthog with anoestrous periods in the European summer. The red river hog was found to be a seasonal and poly oestrous breeder; oestrus cycles started by January and continued until summer. In contrast, the babirusa showed non-seasonal ovarian cyclicity. In pregnant red river hogs, progesterone metabolites were comparable to luteal phase values of the oestrous cycle during the first 3 months of gestation, but did further increase during the last month of pregnancy. Oestrogens and 17-oxo-androstanes were significantly elevated during the second half of gestation. In summary, the reproductive biology of three exotic pig species was studied using non-invasive faecal steroid analysis and these methods were used for comparative investigations of oestrous cycles, pregnancy and seasonality. PMID- 15876500 TI - Development and application of high throughput plasma stability assay for drug discovery. AB - Plasma stability plays an important role in drug discovery and development. Unstable compounds tend to have rapid clearance and short half-life, resulting in poor in vivo performance. This paper examines the variables that affect the plasma stability assay results, including substrate concentration, %DMSO, plasma concentration, enzyme activity upon incubation and batch variation. The results show that plasma stability can accommodate a wide range of experimental conditions. Relatively minor differences in results are produced with major differences in conditions. Significant batch-to-batch variations were observed for rat plasma. We selected the following conditions: 1 microM substrate concentration, 2.5% DMSO, and 50% dilution of plasma in pH 7.4 buffer. Plasma stability can be used as a diagnostic assay when compounds are unexpectedly rapidly cleared, as a special assay when structural classes contain groups that may be susceptible to plasma enzyme hydrolysis, or as general screen for compounds if resources are available. Plasma stability assay has many applications in drug discovery: to alert teams to labile structural motifs, to prioritize compounds for in vivo studies and to screen prodrugs and antedrugs. PMID- 15876501 TI - In vitro anti-rotavirus activity of some medicinal plants used in Brazil against diarrhea. AB - Acute diarrhea, especially in children, is a very common disease with worldwide distribution and with a significant public health impact. Rotaviruses have been recognized as the major agents of diarrhea in infants and young children in developed as well as developing countries. In Brazil, diarrhea is one of the principal causes of death, mainly in the infant population. To fight diarrhea, traditional Brazilian medicine uses a great variety of plants. In this work, 12 medicinal plant species were screened for simian (SA-11) and human (HCR3) rotaviruses inhibition in vitro. At non-cytotoxic concentrations, the extracts from Artocarpus integrifolia L. (Moraceae) bark (480 microg/ml) and Spondias lutea L. (Anacardiaceae) leaves (160 microg/ml) had antiviral activity against both viruses. They showed inhibition of 99.2% and 97%, respectively, for human rotavirus, and 96.4% and 96.2% for simian rotavirus. The extracts from Myristica fragrans Houtt (Myristicaceae) seeds (160 microg/ml) and Spongias lutea bark (40 microg/ml) inhibited human rotavirus (90% and 82.2% inhibition, respectively), whereas the extracts from Anacardium occidentale L. (Anacardiaceae) leaves (4 microg/ml) and Psidium guajava L. (Myrtaceae) leaves (8 microg/ml) showed activity only against simian rotavirus (82.2% and 93.8% inhibition, respectively). Our results indicate that the extracts of Artocarpus integrifolia, Myristica fragrans and Spongias lutea can be useful in the treatment of human diarrhea if the etiologic agent is a rotavirus. PMID- 15876502 TI - A typology of mass grave and mass grave-related sites. AB - Mass graves are archaeological features with humanitarian and forensic import. Their creation and subsequent modification by natural and human agents reflect complex site histories and site formation processes that create a diversity of mass graves that must be captured with adequate terminology. The purpose of this paper is to encourage specialized research within the newly emerging discipline of forensic bioarchaeology of mass grave and mass grave-related sites as they occur internationally. In doing so, the authors present a typology for describing several types of mass grave and mass grave-related sites according to their archaeologically distinctive characteristics. Several definitions are provided to synthesize the experiences of internationally active forensic bioarchaeologists. A series of standardized definitions will ease communication between the forensic bioarchaeology and international human rights communities. We distinguish among the following basic types: surface and grave execution sites, permanent and temporary deposition sites, primary and secondary inhumation sites and, finally, looted instances of the latter. This endeavor is intended to promote communication with legal agencies such as the International Criminal Tribunals (ICTY/ICTR) and International Criminal Court (ICC). PMID- 15876503 TI - Functional MRI in attention-deficit hyperactivity disorder: evidence for hypofrontality. AB - Using event-related functional magnetic resonance imaging to study the Stroop effect on both behavioral and brain activation of ADHD children off or on methylphenidate (MPH). Nine ADHD boys (aged 9.8-14.5 years) and 9 age-matched normal controls were included. A Stroop-like paradigm was used. AFNI (Analysis of Functional NeuroImaging) and its Deconvolution Analysis were used in a descriptive comparison between ADHD and control groups. (1) Both behavioral reaction time and brain activation showed Stroop effect in controls but neither was found in ADHD children off MPH. When MPH was administered, the Stroop effect tended to appear. (2) The activation volume (AV) of prefrontal cortex (PFC) in both the neutral (NC) and interference conditions (IC) in ADHD children off MPH was smaller than in controls. AV of anterior cingulate cortex in the IC in ADHD children off MPH was smaller than that in controls, but was similar in the NC to that in controls. AV of the basal ganglia, insula and cerebellum was also smaller in the IC, but was larger in the NC for ADHD children off MPH compared with controls. These findings are consistent with prior findings of hypofrontality in ADHD children and implicate a compensatory network including basal ganglia, insula and cerebellum for relative lower cognitive load tasks. PMID- 15876504 TI - Autonomic dysfunction in cases of spinal muscular atrophy type 1 with long survival. AB - In Japan, quite a few patients with spinal muscular atrophy type 1 (SMA type 1) survive with mechanical ventilation. Since a patient with SMA type 1 and continuous artificial ventilation exhibited excessive perspiration and tachycardia, we examined the autonomic functions in three cases of SMA type 1, undergoing mechanical ventilation. Two cases exhibited the common sympathetic vagal imbalance on R-R interval analysis involving 24-h Holter ECG recordings in addition to an abnormality in finger cold-induced vasodilatation. Furthermore, one case showed blood pressure and heart rate fluctuation with the paroxysmal elevation, and a high plasma concentration of norepinephrine during tachycardia. These findings suggest that autonomic dysfunction should be examined in SMA type 1 patients with long survival, although the pathogenesis remains to be clarified. PMID- 15876505 TI - [Epidemiology of sexually transmitted infections in France]. AB - BACKGROUND: Sexually transmitted infections (STI) in France are reported on a voluntary basis through several sentinel surveillance systems. METHODS: To monitor STI, sentinel laboratory- or clinician-based surveillance systems were set up by the Institut de Veille Sanitaire: gonorrhea surveillance (Renago) in 1986, Chlamydia infections surveillance (Renachla) in 1989, and more recently, syphilis surveillance in 2000 and rectal lymphogranuloma venereum (LGV) in 2004. RESULTS: From 2000 to 2003, 1,089 syphilis infections were reported. Most of the cases were diagnosed in men having sex with men (MSM) and were mainly reported by STI clinics located in the Paris area. From 1997 to 2000, an increase of gonorrhea was observed each year. After two years of stable trend, the prevalence of gonorrhea increased again in 2003. From 2002 to 2004, 123 LGV cases were diagnosed in France and were observed only in MSM. Since 2001, Chlamydia infections have steadily increased, particularly in women. COMMENTS: Because STI surveillance is based on a voluntary basis, the number of reported cases is probably lower than the number of STI diagnosed in France. However, the data provided by the different surveillance systems reveals that STI have been increasing in France since 1997. Moreover, the resurgence of syphilis in 2000 and the emergence of rectal LGV in 2004 indicate that these STI occur mainly in MSM. Trends on incidence and patients characteristics observed in France are similar to those of several Europeans countries. European Public Health interventions are becoming necessary to prevent and control STI. PMID- 15876506 TI - [Ano-genital lesions due to human papillomavirus infection in women]. AB - Human papillomaviruses (HPV) are the most prevalent sexually-transmitted agents worldwide. HPV are small circular double-stranded DNA epitheliotropic viruses that exhibit either cutaneous or mucosal specificity. Most HPV infections are self-limiting and are spontaneously cleared within months or years. However, infections may persist and result in a variety of benign, pre-malignant and malignant tumors. Cytological and histopathological abnormalities associated with HPV infections of the male and female lower anogenital tract include condylomata, low-grade and high-grade squamous intraepithelial lesions which are incipient cancers, and squamous cell carcinomas. The modal time between HPV infection occurring in the late teens or early 20 s and precancer peaking around 30 years of age is 7-10 years. Women detected with invasive cancers tend to be an average 10 years older than women with high-grade disease. The natural history of cervical cancer reveals that infection with high-risk types may lead to low-grade or high-grade intraepithelial lesions. High-grade lesions may progress to cervical carcinoma if not treated. The purpose of screening, in addition to detecting cervical cancers at an early stage, is to detect and remove high-grade lesions and thus prevent the potential progression to cervical carcinoma. Early detection of cervical neoplasia is possible with regular Pap smears performed from 21 to 70 years of age. In case of abnormal Pap smear, a biopsy performed under colposcopy will allow the diagnosis of cervical lesion. Cancer of the cervix is the second leading cause of cancer related deaths among women across the world (3,400 new cases in France in 2000). PMID- 15876507 TI - [Risk of infection in institutions for elderly people]. AB - The infectious risk in long-term care facilities and nursing homes is significant. Patients living in those facilities are very old, with a poor health status, and a high degree of dependency. The risk for epidemic outbreaks, in particular with viruses, is very high. A simple system for surveillance and action, in relation with hospital infection control units, is mandatory. An educational program is needed to define the prevention program based on the use of hand disinfection and other standard precautions, anti-viral and pneumococcal vaccination. The program must be simple, pragmatic, allowing to maintain social links and quality of life, which are essential for these patients. A strong cooperation between these long-term care facilities and nursing homes, general practitioners, healthcare team, and relatives is necessary. PMID- 15876508 TI - Tissue targeted metabonomics: metabolic profiling by microdialysis sampling and microcoil NMR. AB - The concentration of low molecular weight compounds in tissues can yield valuable information about the metabolic state of an organism. Studies of changes in the metabolic state or metabonomics can reflect disease pathways, drug action, or toxicity. This research aims to develop a new approach, tissue targeted metabonomics. Microdialysis sampling and microcoil NMR analysis are employed to compare basal and ischemic metabolic states of various tissues (blood, brain, and heart) of Sprague-Dawley rats. Microdialysis sampling is localized, making the metabolic profile tissue specific. Coupling to NMR analysis is highly advantageous, because a complete metabolic profile is obtained in a single spectrum. However, small sample volumes and low analyte concentrations make analysis of microdialysis samples challenging. Microcoil NMR uses low sample volumes and has improved mass sensitivity, relative to standard 5 mm probes. The coupling of these techniques is a potentially powerful tool for metabonomics analysis. PMID- 15876509 TI - A validated stability-indicating HPLC with photodiode array detector (PDA) method for the stress tests of Monascus purpureus-fermented rice, red yeast rice. AB - A stability-indicating reversed phase high-performance liquid chromatography (RP HPLC) with photodiode array (PDA) detection method was developed and validated for the assay of monacolin series compounds including monacolin K, L, J and their hydroxyl acid forms as well as dehydroxymonacolin K simultaneously in Monascus purpureus-fermented rice, red yeast rice. Well-resolved peaks of seven main compounds of monacolin family were profiled on a C(18) reverse-phase column using a linear gradient of 0.1% trifluoroacetic acid and acetonitrile as the mobile phase, and the detection wavelength was set at 237nm. The method was validated with respect to specificity, chromatographic parameters, linearity, precision, accuracy, limits of detection and quantitation. The stability stress testing for fermented red yeast rice powder was carried out to show the effects of high temperature (80 degrees C), high humidity at room temperature (92.5% RH, 25 degrees C), high humidity at high temperature (75% RH, 60 degrees C) and light (sunlight) in solid state. The results exhibited that monacolins decreased significantly under the conditions of high humidity at high temperature (75% RH, 60 degrees C) and sunlight. Monacolin K and its hydroxyl acid form would be dehydrolyzed and turned to dehydromonacolin K at high temperature (80 degrees C) while the monacolin K, J and L would be transformed into their corresponding hydroxyl acid forms under the condition of high humidity (92.5% RH, 25 degrees C). The indication is that monacolins in red yeast rice powder are light sensitive and thermal-sensitive. Therefore, it has been suggested that the preparations containing monacolins be stored in the place of cool and lightproof. The proposed degradation pathways were discussed as well. The multi-components assay for stability of botanical products could provide much more information than the normal marker-orientation method. PMID- 15876510 TI - Leptin expression in ruminants: nutritional and physiological regulations in relation with energy metabolism. AB - Leptin, mainly produced in adipose tissue (AT), is a protein involved in the central and/or peripheral regulation of body homeostasis, energy intake, storage and expenditure, fertility and immune functions. Its role is well documented in rodent and human species, but less in ruminants. This review is focused on some intrinsic and extrinsic factors which regulate adipose tissue leptin gene expression and leptinemia in cattle, sheep, goat and camel: age, physiological status (particularly pregnancy and lactation) in interaction with long-term (adiposity) and short-term effects of feeding level, energy intake and balance, diet composition, specific nutrients and hormones (insulin, glucose and fatty acids), and seasonal non-dietary factors such as photoperiod. Body fatness strongly regulates leptin and its responses to other factors. For example, leptinemia is higher after underfeeding or during lactation in fat than in lean animals. Physiological status per se also modulates leptin expression, with lactation down-regulating leptinemia, even when energy balance (EB) is positive. These results suggest that leptin could be a link between nutritional history and physiological regulations, which integrates the animal's requirements (e.g., for a pregnancy-lactation cycle), predictable food availability (e.g., due to seasonal variations) and potential for survival (e.g., body fatness level). Reaching permissive leptin thresholds should be necessary for pubertal or postpartum reproductive activity. In addition to the understanding of leptin yield regulation, these data are helpful to understand the physiological significance of changes in leptin secretion and leptin effects, and how husbandry strategies could integrate the adaptative capacities of ruminant species to their environment. PMID- 15876511 TI - Cell secretion and membrane fusion. AB - Secretion occurs in all cells of multicellular organisms and involves the delivery of secretory products packaged in membrane-bound vesicles to the cell exterior. Specialized cells for neurotransmission, enzyme secretion or hormone release utilize a highly regulated secretory process. Secretory vesicles are transported to specific sites at the plasma membrane, where they dock and fuse to release their contents. Similar to other cellular processes, cell secretion is found to be highly regulated and a precisely orchestrated event. It has been demonstrated that membrane-bound secretory vesicles dock and fuse at porosomes, which are specialized supramolecular structures at the cell plasma membrane. Swelling of secretory vesicles results in a build-up of pressure, allowing expulsion of intravesicular contents. The extent of secretory vesicle swelling dictates the amount of intravesicular contents expelled during secretion. The discovery of the porosome, its isolation, its structure and dynamics at nm resolution and in real time, its biochemical composition and functional reconstitution into artificial lipid membrane, have been determined. The molecular mechanism of secretory vesicle fusion at the base of porosomes, and vesicle swelling, has also been resolved. These findings reveal the molecular mechanism of cell secretion. PMID- 15876512 TI - Endocrinology of milk production. AB - The physiology of lactation includes development of the mammary gland from the foetal to the adult stage, further development during pregnancy and onset of lactation, with the accompanying metabolic and behavioural adaptation. At the onset of pregnancy the endocrine system undergoes dramatic changes. The growth of the mammary gland is stimulated by growth hormone and prolactin, adrenocortical steroids, oestrogens and progesterone, and that of the gastrointestinal (GI) tract by gastrin, CCK and secretin. The onset of lactation is accompanied by increases in the blood volume, cardiac output, mammary blood flow and blood flow through the GI-tract and liver, aiming to provide the udder with nutrients and hormones for regulation of milk synthesis. Food intake and distribution of nutrients to the mammary gland are partially regulated by hormones as well as the repartitioning of nutrients away from body stores towards the udder. To improve milk production, administration of growth hormone has been practised, but also much discussed. Besides central mechanisms, local mechanisms within the mammary gland regulate initiation of lactation, maintenance, regulation of blood flow and mammary gland cell apoptosis. Most of the milk in a filled dairy cow udder is stored in the alveolar compartments. The milk ejection reflex must be activated to gain access to the udder milk, i.e. oxytocin contracts the myoepithelial cells. Recent studies show that vasopressin may also elicit milk ejection. More efficient oxytocin release is achieved if the cows are fed during milking. Beyond milk let down, oxytocin influences maternal behaviour and metabolism. Furthermore, it has been indicated that suckling or milking activates a vagal reflex, which may link the milk production to the endocrine system of the GI tract. The question has been raised whether the mammary gland is a supporting or consuming organ. PMID- 15876513 TI - [Ringer solution: osmolarity and composition revisited]. AB - The composition of Ringer solution, a crystalloid fluid that is often used in anaesthesia and intensive care, varies depending on the manufacturer. The knowledge of the actual content in electrolytes and of the characteristics of this fluid is necessary before it is used. We call attention to a certain Ringer solution (Ringer Maco Pharma, Maco Pharma), for which the manufacturer's information about the tonicity and the osmolarity was incorrect. Contrary to what is written on the bag and in the product description (isotonicity, osmolarity of 276.8 mOsm/l), the theoretical osmolarity was 221.4 mOsm/l and the measured osmolality was about 208 mmol/kg, exposing the hypotonic characteristics of this fluid. The use of this product is potentially dangerous in patients with pathologies where the infusion of free water is especially badly supported. PMID- 15876514 TI - [Organisation of care for patients suffering from subarachnoid haemorrhage]. PMID- 15876515 TI - [Sedation in ICU: from clinical research to daily practice]. PMID- 15876516 TI - Isolated idiopathic hypomagnesemia presenting as aphasia and seizures. AB - Isolated hypomagnesemia of the idiopathic form is a rare condition that is known to present as generalized motor seizures in children. This report describes a 4 year-old African-American male who presented with a predominant symptom of sudden onset aphasia and no clear initial motor seizure activity. An evaluation revealed an isolated and severe hypomagnesemia (initial serum magnesium levels <1.0 mg/dL) and inappropriate renal handling of magnesium (fractional excretion of magnesium >40% under conditions of hypomagnesemia). The child had subsequent generalized tonic-clonic seizures that were brought under control with valproic acid therapy and magnesium supplementation. Six months after the diagnosis, he had regained 50 60% of his speech and had no further staring spells or motor seizure activity after the initial episode. Isolated and idiopathic hypomagnesemia caused by defective renal reabsorption of magnesium is a rare familial condition with variable inheritance. Aphasia as the solitary presenting symptom has not been described before. The exact pathophysiology of hypomagnesemic aphasia and seizures is not known but may relate to disinhibition of specific types of glutamate receptors. In the present case, neuronal depolarization may have been localized to language areas in the temporal lobes. PMID- 15876517 TI - A female with Angelman syndrome and unusual limb deformities. AB - This report presents the case of a 13-year-old female with Angelman syndrome caused by 15q11-13 microdeletion demonstrating unusual marked limb deformities with generalized osteoporosis, delayed bone age, and brachydactyly type B. The radiographs of her femur, tibia, fibula, ulna, and radius revealed curved deformities in the distal diaphysis-metaphysis areas and generalized osteoporosis. This can be explained by the patient's severe disability, delayed puberty, presumed nutritional and environmental deficits, or rickets. In addition, she had shortening of the distal phalanges of all fingers, the absence of some epiphyses of the distal phalanges, and hypertrophic and curved III metacarpal bones. These clinical findings could not be explained by classical rickets or osteoporosis, but can be classified as brachydactyly type B. To our knowledge, such marked limb deformities and brachydactyly have not previously been described in patients with Angelman syndrome. PMID- 15876518 TI - Use of zonisamide in pediatric patients. AB - Zonisamide is a broad-spectrum antiepilepsy drug approved in the United States for the adjunctive treatment of partial seizures in adults with epilepsy. Studies in Japan have established zonisamide as effective and safe for use in children, but there is limited information from the United States concerning its use in pediatric patients. This chart-review study was conducted to evaluate the efficacy, safety, and appropriate dosage of zonisamide for treating seizures in pediatric and adolescent patients. Charts of 50 pediatric patients (mean age 9.1 years, range 9 months to 20 years) who received zonisamide were evaluated for demographic characteristics, seizure types, dosage (mean = 15.8 mg/kg/day), response, concurrent medications, and adverse events. After treatment, 8 patients were seizure-free, and 11 others had > or =50% improvement in seizure control, including 11 of the 28 patients for whom treatment with six or more antiepileptic drugs was insufficient. Thirty-one patients experienced at least one adverse event while receiving zonisamide, and 14 discontinued as a result. PMID- 15876519 TI - Management of tectal glioma in childhood. AB - Tectal glioma is a topographical diagnosis including tumors of different histology, mainly low-grade astrocytomas. Clinical symptoms are usually associated with increased intracranial pressure. This report discusses the management of this rare tumor in children. Clinical charts of 12 children with tectal glioma treated in our department between 1976 and 2001 were retrospectively reviewed. The mean age at the time of diagnosis was 6.75 years (range, 4 weeks to 16 years). The duration between first symptoms and the diagnosis of tectal glioma was in the range of 2 days to 9 years. Ten patients presented with symptoms associated with increased intracranial pressure, one patient presented with ataxia, and in one case tectal glioma was an incidental finding. First-line therapy was endoscopic third ventriculostomy in 5 cases (42%), ventriculoperitoneal shunting in 6 cases (50%), and combined partial tumor resection and shunting in one case. Histology was obtained in 5 cases (low-grade astrocytoma, n = 4; ependymoma, n = 1). All patients had good neurologic function at the end of follow-up. Tectal glioma represents a distinct subgroup of brainstem tumors associated with a good (or favorable) prognosis. Effective treatment for hydrocephalus is essential; the tumor should be monitored by regular clinical examination and magnetic resonance imaging. Biopsy is warranted in cases with tumor progression. PMID- 15876520 TI - Early-onset ataxia with oculomotor apraxia with a novel APTX mutation. AB - Early-onset ataxia with oculomotor apraxia and hypoalbuminemia is an autosomal recessive cerebellar ataxia characterized by oculomotor apraxia, peripheral neuropathy, and hypoalbuminemia. Mutations in aprataxin gene located at chromosome 9q13 have been identified recently in Japanese and European patients. This study reports two cases of siblings with early-onset ataxia with oculomotor apraxia and hypoalbuminemia, which manifested early onset before 2 years of age with relatively rapid progression and severe dystonia. Both of the siblings were compound heterozygotes with aprataxin gene mutations, 689 insT and G692A, in exon 5 that encodes the histidine triad domain of the aprataxin protein. The novel missense mutation, G692A, was not present in 40 unrelated and unaffected individuals. PMID- 15876521 TI - Limb preference in children with obstetric brachial plexus palsy. AB - Brachial plexus palsy affects children differently than adults. In children with obstetric brachial plexus palsy, motor development must depend on nervous system adaptation. Previous studies report sensory plasticity in these children. This noninvasive study provides support for neural plasticity (the general ability of the brain to reorganize neural pathways based on new experiences) in children with obstetric brachial plexus palsy by considering upper limb preference. As in the general population, we expect that 90% of children would prefer their right upper limb. However, only 17% of children affected by right obstetric brachial plexus palsy prefer the right upper limb for overall movement; children with left obstetric brachial plexus palsy did not significantly differ from the general population in upper limb preference. This study also provides the first evidence of a significant correlation between actual task performance and select obstetric brachial plexus palsy outcome measurement systems, thereby justifying the routine use of these outcome measurement systems as a reflection of the practical utility of the affected limb to the patient. PMID- 15876522 TI - T-cell subsets and interleukin-6 response in Rasmussen's encephalitis. AB - To evaluate the immunopathogenesis in Rasmussen's encephalitis, peripheral lymphocyte subsets and interleukin-6 analysis were performed in three patients. Magnetic resonance spectroscopy and diffusion-weighted magnetic resonance imaging were performed to assess neuronal injury in the affected hemisphere. Before initiation of immune therapy, percentage of cytotoxic T cells was found to be increased in peripheral blood obtained from patients compared with a group of age matched normal control subjects. During follow-up, percentage of cytotoxic T cells returned to the normal ranges only in one patient who had an early functional hemispherectomy. All three patients had significantly increased interleukin-6 concentration in cerebrospinal fluid and serum compared with the mean values of patients with acute viral encephalitis. The magnitude of interleukin-6 response in the patients correlated with the neuronal loss and atrophy on magnetic resonance spectroscopy and diffusion-weighted magnetic resonance imaging studies. The patient, who had a fulminant course and an early hemispherectomy, had higher interleukin-6 concentration in cerebrospinal fluid and serum than those of the other two. Detection of an increased percentage of cytotoxic T cells in peripheral blood supports the presence of a T cell-mediated inflammatory pathogenesis in Rasmussen's encephalitis. However, elevated interleukin-6 response might reflect the magnitude of the inflammatory process in the affected hemisphere. PMID- 15876523 TI - Executive functions after traumatic brain injury in children. AB - There is growing recognition that executive function, the superordinate, managerial capacity for directing more modular abilities, is frequently impaired by traumatic brain injury in children and mediates the neurobehavioral sequelae exhibited by these patients. This review encompasses the definition of specific executive functions, age-related changes in executive functions in typically developing children, and the effects of traumatic brain injury on executive functions. The neural substrate for executive functions is described, including relevant functional brain imaging studies that have implicated mediation by prefrontal and parietal cortex and their circuitry. The vulnerability of the neural substrate for executive function to the pathophysiology of traumatic brain injury is discussed, including focal lesions and diffuse axonal injury. Domains of executive functions covered in this review include the basic processes of working memory and inhibition and more complex processes such as decision making. Other domains of executive function, including motivation, self-regulation, and social cognition are discussed in terms of research methodology, clinical assessment, and findings in children with traumatic brain injury. Proposed approaches to the rehabilitation of executive functions are presented. PMID- 15876524 TI - The effect of the antibody used and method of quantification on oral mucosal vascularity. AB - A vast literature exists on angiogenesis particularly with regard to understanding tumour growth and development. This is no less true for oral cancer, where slight variations in protocols may invalidate comparison between studies. The influence of tissue processing, and the specific antibody and method of quantification used to identify blood vessels was assessed with respect to oral mucosal vascularity. The angiogenic profile of 20 frozen sections of clinically normal oral mucosa were compared with 16 paraffin-embedded specimens. Eight of the frozen specimens had matched paraffin-embedded tissue. Vascular antigens were identified immunohistochemically using four antibodies and blood vessel density was measured by three methods. CD31 and von Willebrand's factor (vWF) counts were higher than alpha-smooth muscle actin and alphanubeta3. Results for CD31 were higher in frozen tissue than paraffin-embedded tissue. Significantly more CD31-positive vessels were found in frozen tissue in the cohort of matched samples. For paraffin-embedded tissue vWF gave higher values for vascularity than CD31. The variation in antibody reaction to these markers of blood vessels suggests that the antibody employed and method of fixation is more important than the method of quantification used. Comparison of the results for fresh frozen tissue and paraffin-processed tissue suggests that this is acceptable for vWF but not for CD31. PMID- 15876525 TI - Long-term clinical and radiological outcomes of surgical management of central giant cell granuloma of the maxilla. AB - The aim of this study was to report the results of long-term clinical and radiological follow up of the surgical management of central giant cell granuloma of the maxilla. A retrospective analysis was conducted on 12 patients--5 male and 7 female whose ages ranged from 9 to 41 years, with central giant cell granuloma of the maxilla treated between 1991 and 2000. The anterior maxilla was involved in five cases, seven cases were in the posterior region, and none crossed the midline. Four patients were asymptomatic and eight patients presented with pain, mobility of teeth, and rapidly enlarging facial swelling. Radiographically, all lesions were radiolucent; 58.3% were unilocular, and the borders were well defined in 33.3%. Roots were commonly displaced by the lesion, but no root resorption was observed. Cortical perforation was seen clinically and on CT scan in four patients. All cases were treated with thorough curettage until healthy bone was encountered. In cases with cortical perforation the affected buccal and/or palatal mucosa was excised. When the maxillary sinus was affected or the lesion was bulging into the nose, the nasal mucosa was also removed. All patients were reviewed annually for a follow-up period of 2-9 years; the condition recurred in two patients. It is concluded that thorough curettage is a satisfactory method for the treatment of non-aggressive and aggressive central giant cell granuloma of the maxilla, with a low recurrence rate and favorable postoperative outcome. PMID- 15876526 TI - A double mutation in the gyrA gene is necessary to produce high levels of resistance to moxifloxacin in Campylobacter spp. clinical isolates. AB - The aim of this study was to compare different fluoroquinolones against Campylobacter spp., analysing the molecular mechanisms of resistance. Moxifloxacin exhibited the greatest activity of the quinolones tested, being active against isolates carrying a single mutation in the gyrA gene. High resistance levels to moxifloxacin were related to the presence of a double gyrA mutation. PMID- 15876527 TI - Dynamic changes of anti-oxidative enzymes of 10 wheat genotypes at soil water deficits. AB - Drought is a world-spread problem seriously influencing crop production and quality, the loss of which is the total for other natural disasters, with increasing global climate change making the situation more serious. Wheat is the staple food for more than 35% of world population, so wheat anti-drought physiology study is of importance to wheat production and biological breeding for the sake of coping with abiotic and biotic conditions. Much research is involved in this hot topic, but the pace of progress is not so large because of drought resistance being a multiple-gene-control quantitative character and wheat genome being larger (16,000 Mb). On the other hand, stress adaptive mechanisms are quite different, with stress degree, different growth and developmental stages, time course, materials and experimental plots, thus increasing the complexity of the issue in question. Additionally, a little study is related to the whole life circle of wheat, which cannot provide a comprehensive understanding of its anti drought machinery. We selected 10 kinds of wheat genotypes as materials, which have potential to be applied in practice, and measured change of relative physiological indices through wheat whole growing developmental circle (i.e. seedling, tillage and maturing). Here, we reported the dynamic anti-oxidative results of whole stage (i.e. seedling, tillage and maturing) in terms of activities of POD, SOD, CAT of 10 wheat genotypes as follows: (1) 10 wheat genotypes can be grouped into three kinds (A, B and C, respectively) according to their changing trend of the measured indices; (2) A group performed better resistance drought under the condition of treatment level 1, whose activities of anti-oxidative enzymes (POD, SOD, CAT) were higher; (3) B group exhibited stronger anti-drought under treatment level 2, whose activities of anti-oxidative enzymes were higher; (4) C group expressed anti-drought to some extent under treatment level 3, whose activities of anti-oxidative enzymes were stronger, MDA lower; (5) these results demonstrated that different wheat genotypes have different physiological mechanisms to adapt themselves to changing drought stress, whose molecular basis is discrete gene expression profiling (transcriptom); (6) our results also showed that the concept and method accepted and adopted by most researchers--that 75% FC is a proper supply for higher plants -was doubted because this level could not reflect the true suitable level of different wheat genotypes; (7) our research can provide insights into physiological mechanisms of crop anti-drought and direct practical materials for wheat anti-drought breeding; (8) POD, SOD and CAT activities of different wheat genotypes had quite different changing trend at different stages and under different soil water stress conditions, which was linked with their origin of cultivation and individual soil water stress threshold; (9) our primary results also firstly displayed that the changing trend for wheat adapting to environmental stress during life circle was an S-shaped curve, which is, by chance, consistent with Plant Growth Grand Periodicity Curve. PMID- 15876528 TI - Steady-state haemoglobin level in sickle cell anaemia increases with an increase in erythrocyte membrane n-3 fatty acids. AB - The aim of the study was to investigate, whether (a) patients with homozygous sickle cell disease (SCD, HbSS) have abnormal blood fatty acids; (b) the abnormality, if it exists, affects all the plasma and erythrocyte lipids or it is restricted to a particular lipid moiety; (c) there is an association between levels of membrane n-3 or n-6 long-chain polyunsaturated fatty acids (LCPUFA) and the degree of anaemia. Fatty acids of erythrocyte choline (CPG), serine (SPG) and ethanolamine (EPG) phosphoglycerides and sphingomyelin (SPM); and plasma CPG, triglycerides and cholesterol esters of 43 steady-state HbSS patients and 43 ethnically matched, healthy, HbAA controls were analysed. The levels of the n-6 LCPUFA, arachidonic (AA), adrenic and docosapentaenoic acids in erythrocyte CPG (P<0.001) and EPG (P<0.01) were higher in the patients compared with the controls. In contrast, the proportions of eicosapentaenoic acid (EPA) in CPG and EPG (P<0.001) and docosahexaenoic acid (DHA) and total n-3 metabolites in CPG (P<0.001) were lower in the patients. The steady-state haemoglobin level of the patients correlated with erythrocyte DHA (r=0.55, P<0.01), EPA (r=0.38, P<0.05) and total n-3 metabolites (r=0.51, P<0.001) in CPG. Also, it correlated with erythrocyte EPA (r=0.64, P<0.01) and total n-3 metabolites (r=0.42, P<0.01) in EPG. The study revealed an imbalance between n-3 and n-6 LCPUFA in erythrocyte and plasma lipid moieties of the HbSS group. Furthermore, it suggested that correction of the imbalance by supplementation with EPA and DHA could ameliorate anaemia in the patients. This observation is consistent with the results of pilot studies, which demonstrated that treatment with n-3 fatty acids confers clinical benefit to sickle cell patients. PMID- 15876529 TI - Brain aromatase in Japanese medaka (Oryzias latipes): Molecular characterization and role in xenoestrogen-induced sex reversal. AB - In female fish estrogen is required for the development of primary and secondary sex characteristics and is derived from the aromatization of androgens by aromatase. There are two isoforms of aromatase in several teleost species, brain and ovarian. The objective of this study was two-fold: clone and sequence the coding and promoter region of brain aromatase in medaka, and determine the effects of exposure to an environmental estrogen (o,p-DDT) on sex determination and brain aromatase transcription and activity. The brain aromatase coding sequence was obtained by reverse transcription polymerase chain reaction (RT-PCR) and PCR-based genomic DNA walking was used to clone the promoter of the brain aromatase gene. The promoter sequence revealed potential binding sites for the estrogen receptor and for transcription factors involved in primary neurogenesis and sex determination. Medaka fry were exposed to increasing o,p-DDT concentrations (0-5.5 microg/L) from days 1 to 15 after hatch and brain aromatase expression and activity were measured on days 5, 9, and 14. A complete male-to female sex reversal occurred at 5.5 microg/L o,p-DDT and aromatase activity and expression data showed a significant five-fold increase at this concentration at day 14. This information suggests that brain aromatase is involved in the abnormal sexual differentiation of fish treated with xenoestrogens. PMID- 15876530 TI - Design and synthesis of new 2-substituted-5-(2-benzylthiophenyl)-1,3,4 oxadiazoles as benzodiazepine receptor agonists. AB - A series of new 2-substituted-5-(2-benzylthiophenyl)-1,3,4-oxadiazoles was designed and synthesized as anticonvulsant agents. Conformational analysis and superimposition of energy minima conformers of the designed molecules on estazolam, a known benzodiazepine receptor agonist, revealed that the main proposed benzodiazepine pharmacophores were well matched. Electroshock and pentylenetetrazole-induced lethal convulsion tests showed that the introduction of an amino group in position 2 of 1,3,4-oxadiazole ring and a fluoro substituent at para position of benzylthio moiety had the best anticonvulsant activity. It seems this effect is mediated through benzodiazepine receptors mechanism. PMID- 15876531 TI - A new orally bioavailable dual adenosine A2B/A3 receptor antagonist with therapeutic potential. AB - The synthesis and SAR of 5-heterocycle-substituted aminothiazole adenosine receptor antagonists is described. Several compounds show high affinity and selectivity for the A2B and A3 receptors. One compound (5f) shows good ADME properties in the rat and as such may be an important new compound in testing the current hypotheses proposing a therapeutic role for a dual A2B/A3 antagonist in allergic diseases. PMID- 15876532 TI - Synthesis, antitumor evaluation and DNA photocleaving activity of novel methylthiazonaphthalimides with aminoalkyl side chains. AB - A series of methylthiazonaphthalimides was synthesized and quantitatively evaluated as efficient DNA intercalators, antitumor agents and DNA photocleavers. A(1) showed both efficient antitumor activities against cell lines of A549 and P388 with IC50 of 82.8 and 31 nM, respectively. A(3) was the strongest antitumor agent against A549 with the IC50 of 20.8 nM. A(2), the most efficient DNA intercalator, was found to be the strongest DNA photocleaver via superoxide anion. An explanation was given for the disaccord between antitumor and DNA photocleaving activities. PMID- 15876533 TI - Use of PROTACS as molecular probes of angiogenesis. AB - Small molecules designed to specifically activate or inactivate protein functions have been useful to study biological processes. PROTACS are small molecule chimera which comprise a ligand and a peptide recognition motif for an E3 ligase. These novel reagents exploit the ubiquitin-mediated proteasome degradation pathway to target the ligand-bound protein for intracellular degradation. Here, we report that an estrogen receptor (ER)-targeting PROTACS that causes degradation of ER is able to potently inhibit endothelial cell differentiation in a three-dimensional angiogenic sprouting assay. These findings support the use of ER-targeting PROTACS as probes of angiogenesis. PMID- 15876534 TI - Synthesis of all-trans arachidonic acid and its effect on rabbit platelet aggregation. AB - A simple and high-yielding method to convert natural all-cis PUFA derivatives to the corresponding all-trans geometrical isomers is described. The method is based on the thiyl radical-catalyzed cis-trans isomerization. The all-trans isomer of arachidonic acid was found to cause rabbit platelet aggregation at concentrations higher than 0.1 mM and inhibition of PAF-induced platelet aggregation in a concentration dependent manner with an IC(50) in the micromolar range. PMID- 15876535 TI - Estrogen receptor ligands: design and synthesis of new 2-arylindene-1-ones. AB - The syntheses of a series of 2-arylindene-1-ones as potent ligands of ERbeta and ERalpha are described. Several compounds exhibited high potency and moderate selectivity for the ERbeta receptor. X-ray and modeling studies were used to understand ligand binding orientation and observed affinity. PMID- 15876536 TI - Sequence specific recognition of DNA by tailor-made hairpin conjugates of achiral seco-cyclopropaneindoline-2-benzofurancarboxamide and pyrrole-imidazole polyamides. AB - Hairpin conjugates of achiral seco-cyclopropaneindoline-2-benzofurancarboxamide (achiral seco-CI-Bf) and three diamides (ImPy 1, PyIm 2, and PyPy 3, where Py is pyrrole, and Im is imidazole), linked by a gamma-aminobutyrate group, were synthesized. The sequence-specific covalent alkylation of the achiral CI moiety with adenine-N3 in the minor groove was ascertained by thermally induced DNA cleavage experiments. The results provide evidence that hairpin conjugates of achiral seco-CI-Bf-gamma-polyamides could be tailored to target specific DNA sequences according to a set of general rules: the achiral CI moiety selectively reacts with adenine-N3, a stacked pair of imidazole/benzofuran prefers a G/C base pair, and a pyrrole/benzofuran prefers an A/T or T/A base pair. Models for the binding of hairpin conjugates 1-3 with sequences 5'-TCA(888)G-3', 5'-CAA(857)C 3', and 5'-TTA(843)C-3' are proposed. PMID- 15876537 TI - Synthesis of carbamate-linked lipids for gene delivery. AB - Series of lipids 1a-d and 2a,b, with carbamate linkages between hydrocarbon chains and ammonium or tertiary amine head, which were pH sensitive, were synthesized for liposome-mediated gene delivery. The variable length of carbon chains and quaternary ammonium or neutral tertiary amine heads allowed to find the structure-function relationship of how these factors affect cationic lipids on gene delivery performance. PMID- 15876538 TI - Studies on quinones. Part 39: Synthesis and leishmanicidal activity of acylchloroquinones and hydroquinones. AB - Acylhydroquinone-based compounds are attractive targets for the design of new leishmanicidal drugs. We have previously described sesquiterpene quinones and hydroquinones series, which exhibit different degree of potency against Leishmania amazonensis. The present study details the preparation of acylchloroquinones and hydroquinones possessing lipophilic substituents and examines their in vitro activity against intracellular L. amazonensis amastigotes. The quinone or hydroquinone nucleus is essential for the activity of the members of the series. The lipophilicity of the cycloaliphatic systems in these members seems to attenuate the cytotoxical effect and increases the selectivity of those compounds containing the norbornene system. PMID- 15876539 TI - Docking simulation with a purine nucleoside specific homology model of deoxycytidine kinase, a target enzyme for anticancer and antiviral therapy. AB - 5'-Phosphorylation, catalyzed by human deoxycytidine kinase (dCK), is a crucial step in the metabolic activation of anticancer and antiviral nucleoside antimetabolites, such as cytarabine (AraC), gemcitabine, cladribine (CdA), and lamivudine. Recently, crystal structures of dCK (dCKc) with various pyrimidine nucleosides as substrates have been reported. However, there is no crystal structure of dCK with a bound purine nucleoside, although purines are good substrates for dCK. We have developed a model of dCK (dCKm) specific for purine nucleosides based on the crystal structure of purine nucleoside bound deoxyguanosine kinase (dGKc) as the template. dCKm is essential for computer aided molecular design (CAMD) of novel anticancer and antiviral drugs that are based on purine nucleosides since these did not bind to dCKc in our docking experiments. The active site of dCKm was larger than that of dCKc and the amino acid (aa) residues of dCKm and dCKc, in particular Y86, Q97, D133, R104, R128, and E197, were not in identical positions. Comparative docking simulations of deoxycytidine (dC), cytidine (Cyd), AraC, CdA, deoxyadenosine (dA), and deoxyguanosine (dG) with dCKm and dCKc were carried out using the FlexX docking program. Only dC (pyrimidine nucleoside) docked into the active site of dCKc but not the purine nucleosides dG and dA. As expected, the active site of dCKm appeared to be more adapted to bind purine nucleosides than the pyrimidine nucleosides. While water molecules were essential for docking experiments using dCKc, the absence of water molecules in dCKm did not affect the ability to correctly dock various purine nucleosides. PMID- 15876540 TI - Synthesis of a novel potent cyclic peptide MC4-ligand by ring-closing metathesis. AB - The synthesis of a novel potent cyclic peptide MC4-ligand by ring-closing metathesis (RCM) is described. Based on the Ac-Nle-Gly-Lys-D-Phe-Arg-Trp-Gly-NH2 MC4 ligand, Ac-Nle-Alg-Lys-D-Phe-Arg-Trp-Alg-NH2 was designed and synthesized followed by cyclization using RCM. Both compounds are high affinity and selective MC4-R-agonists. The cyclic RCM-peptide was more potent in a rat-grooming assay. PMID- 15876541 TI - Fibrin sealant treatment of splenic injuries during oesophagectomy. AB - The incidence of 'accidental' splenectomy during oesophagectomy varies from 4 to 42% and leads to an increase in sepsis, anastomotic leakage and mortality. Therefore, preservation of the spleen should be the aim of treatment whenever splenic injury occurs. We describe the use of fibrin glue delivered as a thin film by a spray applicator for rapid and definitive haemostasis of grades 1 and 2 splenic injuries in patients undergoing oesophagectomy. PMID- 15876542 TI - Resolving tylenchid evolutionary relationships through multiple gene analysis derived from EST data. AB - Sequence-based phylogenetic analyses typically are based on a small number of character sets and report gene trees which may not reflect the true species tree. We employed an EST mining strategy to suppress such incongruencies, and recovered the most robust phylogeny for five species of plant-parasitic nematode (Meloidogyne arenaria, M. chitwoodi, M. hapla, M. incognita, and M. javanica), three closely related tylenchid taxa (Heterodera glycines, Globodera pallida, and G. rostochiensis) and a distant taxon, Caenorhabditis elegans. Our multiple-gene approach is based on sampling more than 80,000 publicly available tylenchid EST sequences to identify phylum-wide orthologues. Bayesian inference, minimum evolution, maximum likelihood and protein distance methods were employed for phylogenetic reconstruction and hypothesis tests were constructed to elucidate differential selective pressures across the phylogeny for each gene. Our results place M. incognita and M. javanica as sister taxa, with M. arenaria as the next closely related nematode. Significant differences in selective pressure were revealed for some genes under some hypotheses, though all but one gene are exclusively under purifying selection, indicating conservation across the orthologous groups. This EST-based multi-gene analysis is a first step towards accomplishing genome-wide coverage for tylenchid evolutionary analyses. PMID- 15876543 TI - Outcomes in newly diagnosed localization-related epilepsies. AB - A total of 558 patients with a range of localization-related epilepsy syndromes starting treatment in a single centre were followed over a period of up to 20 years. Overall, 343 (62%) patients became seizure free for 12 months or more (responders), 92% of whom (57% of total population) remained in remission until the end of follow-up. Only 27 (5%) responders relapsed and subsequently developed refractory epilepsy. The remaining 215 (38%) patients never became seizure free for any 12-month period. There were no significant differences in outcome between cryptogenic (56% remission) and symptomatic (57% remission) epilepsies. Patients with underlying cortical atrophy (71% remission; p<0.05) or cerebrovascular disease (70% remission; p<0.01) did better, while those with traumatic brain injury (35% remission; p<0.001) did worse than the remainder of the symptomatic group. Remission rates in patients with cortical dysplasias (60%), hippocampal atrophy (50%) and primary brain tumors (52%) appeared no different from those with other symptomatic epilepsies. Overall, 20-40% patients with each epilepsy syndrome reported no further seizures after starting AED treatment including 21% with hippocampal atrophy and 33% with cortical dysplasia. More than 50% of patients developing localization-related epilepsy during adolescence or in adulthood had a good outcome. Prognosis in those with underlying hippocampal atrophy or cortical dysplasia was not always bad. PMID- 15876544 TI - Risk factors for carbamazepine elevation and toxicity following epilepsy surgery. AB - A proportion of patients undergoing epilepsy surgery, and receiving carbamazepine (CBZ), experience significant elevations in CBZ plasma concentrations, some with associated CBZ toxicity. The objective of this study was to identify significant risk factors for elevations (>12 microg/ml) in CBZ concentrations and CBZ-induced toxicity following epilepsy surgery. METHODS: We retrospectively examined charts of 74 inpatients (31 children and 43 adults) chronically receiving CBZ and undergoing epilepsy surgery between January 1996 and June 2000. Patient demographics, medications, type of surgery, seizure history, adverse events, CBZ doses and concentrations were evaluated. RESULTS: 51.2% of adults and 51.6% of pediatric patients had drug elevations. In the pediatric group, 12.9% had symptoms of toxicity compared to 9.3% in the adult group. Five risk factors-pre operative CBZ dose, fentanyl dose, surgery day CBZ concentration, body weight, and blood loss-were related to post-operative CBZ concentrations. Three risk factors: age <18 years, pre-operative CBZ dose, and the surgery day CBZ (immediate pre-operative) concentration, were significantly related to the outcome measure of abnormal CBZ concentration (>12 microg/ml). Two variables significantly related to toxicity were average post-operative CBZ dose and the surgery day CBZ concentration. Increases in concentrations occurred at a mean 33+/-13.7 h (range: 11-74 h) after surgery. DISCUSSION: Based upon our results in patients with one or more risk factors, we suggest that reduction of post operative CBZ doses be considered. PMID- 15876545 TI - Cell-binding properties of the envelope proteins of porcine endogenous retroviruses. AB - To examine the binding properties of the envelope glycoproteins of porcine endogenous retrovirus subgroups A and B (PERV-A and PERV-B), we produced two forms of soluble envelope proteins, termed Env-ST and Env-SU, using a baculovirus expression system. Env-ST and Env-SU encompass one-third of the N-terminal and the entire surface unit (SU) of the envelope protein, respectively. Using these proteins, binding assays were performed in various mammalian cell lines. The binding properties of the Env-STs that contain the putative receptor binding domain (RBD) did not correlate with the susceptibility to the pseudotype viruses having PERV envelopes, whereas those of the Env-SUs correlated fairly well. These results suggested that the Env-SUs but not Env-STs interacted with their receptors in various cell lines. Interestingly, PERV-A Env-SU did not bind to a mink cell line (Mv1-Lu cells) that is highly susceptible to the PERV-A pseudotype virus. In addition, PERV-B Env-SU did not interfere with the PERV-B pseudotype virus on Mv1-Lu cells. These results suggest the existence of a cognate receptor independent entry pathway as demonstrated in an immunodeficiency-inducing variant of feline leukemia virus FeLV. PMID- 15876546 TI - The cytolethal distending toxin of Haemophilus ducreyi aggravates dermal lesions in a rabbit model of chancroid. AB - Haemophilus ducreyi, the etiologic agent of the sexually transmitted disease chancroid, produces a cytolethal distending toxin (HdCDT) that inhibits cultured cell proliferation, leading to cell death. A rabbit model of dermal infection was used to investigate the roles of H. ducreyi bacteria and HdCDT in the development, clinical appearance, and persistence of infection. A non-toxin producing H. ducreyi strain, and for comparison purposes a non-capsulated Haemophilus influenzae strain, were inoculated intradermally, with and without co administration of purified HdCDT. Co-administration of HdCDT resulted in significant aggravation of H. ducreyi-induced inflammatory lesions, and development of ulcers in rabbit skin. Less pronounced inflammatory lesions and lack of epithelial eruption were observed after inoculation with H. influenzae. Histopathological sections of the H. ducreyi-induced lesions, in both the presence and absence of HdCDT, showed dense infiltrates of the same type inflammatory cells, with the exception of a prominent endothelial cell proliferation noted in sections from lesions caused by H. ducreyi and toxin. Signs of chronic inflammation with involvement of T cells, macrophages, eosinophils, and granuloma formation were observed after H. ducreyi inoculation both with and without toxin. In conclusion, H. ducreyi causes a pronounced, chronic inflammation with involvement of T cells and macrophages, and in combination with HdCDT production of ulcers in the rabbit model. These pathogenic mechanisms may promote the development and persistence of chancroid ulcers. PMID- 15876547 TI - A tissue explant system for assessing tendon overuse injury. AB - Tendon overuse injuries are common athletic and occupational problems. When studying mechanisms that cause these injuries, inherent complexities associated with controlling in vivo loading necessitates alternative approaches such as in vitro organ culture. Current devices for loading explants in organ culture, whether custom-built or commercial, have various deficiencies in their loading capability, control mechanism and strain assessment. To overcome these shortcomings, an advanced tissue loading device with video strain analysis capabilities was developed for investigating overuse injuries and its performance/calibration were evaluated. Two tests were used to assess the ability of the system to create and monitor mechanical changes with overuse. Overuse loading significantly increased strains and decreased strength, showing the ability of this system to create and monitor tissue damage. Furthermore, the device design allows for its use in a standard incubator. Coupled with custom loading and data collection programs, this system is suitable for long-term overuse injury studies. PMID- 15876548 TI - Genotoxic damage in mine workers exposed to diesel exhaust, and the effects of glutathione transferase genotypes. AB - This study was performed in an Estonian shale-oil mine with the purpose to develop and apply a number of biomarkers for occupational diesel-exhaust exposure monitoring. Increased breathing-zone exposures to exhaust from operators of diesel-powered trucks in the mine was confirmed in the environmental monitoring part of the study, showing a 7.5-fold higher exposure to particle-associated 1 nitropyrene (1-NP) in 50 underground workers compared with 42 surface workers [P.T.J. Scheepers, D. Coggon, L.E. Knudsen, R. Anzion, H. Autrup, S. Bogovski, R.P. Bos, D. Dahmann, P. Farmer, E.A. Martin, V. Micka, V. Muzyka, H.-G. Neumann, J. Poole, A. Schmidt-Ott, F. Seiler, J. Volf, I. Zwirner-Baier, Biomarkers for occupational diesel exhaust exposure monitoring (BIOMODEM)-a study in underground mining, Toxicol. Lett. 134 (2002) 305-317; P.T.J. Scheepers, V. Micka, V. Muzyka, R. Anzion, D. Dahmann, J. Poole, R.P. Bos, Exposure to dust and particle associated 1-nitropyrene of drivers of diesel-powered equipment in underground mining, Ann. Occp. Hyg. 47 (2003) 379-388]. Analysis of DNA damage by the Comet assay on frozen blood samples was performed on the total study group and showed significantly higher levels (p=0.003) in underground workers (smokers) driving diesel-powered excavation machines (median 155 on a scale from 0 to 400, among 47 persons), compared with surface workers who smoked (median of 90, among 46 persons). The level of DNA damage in underground smokers was significantly higher (p=0.04) than in non-smokers. Samples from 2 of the 3 sampling weeks had significantly lower DNA damage compared with the third week, probably due to timely processing and freezing. These samples also showed significant differences (p<0.001) between underground workers (median 145, among 41 persons) and surface workers (median 60, among 30 persons). An HPLC method was developed for the analysis of (32)P-postlabelled 1-NP-DNA-adducts, and was applied to a sub-sample of 20 workers. No significant differences between surface and underground workers were found in this sub-sample with respect to the minor, unidentified adducts that had similar chromatographic properties to 1-NP adducts, and smoking did not have any effect on adduct levels. No significant effects of the genotypes of GSTM1, GSTP1 and GSTT1 on DNA-adducts and on DNA damage as measured by the Comet assay were found in the total study group. The study confirms an increased level of DNA damage in workers exposed to exhaust from truck-driving in the mine. However, the results of the environmental and biological monitoring of 1-NP did not correlate, suggesting that inhalation exposure to diesel exhaust is not reflected by an increase in 1-NP-DNA-adduct levels and/or that factors other than occupational exposure to diesel exhaust are primary determinants of these DNA adduct levels. PMID- 15876549 TI - Synthesis, spectroscopy and electrochemical behaviors of nickel(II) complexes with tetradentate shiff bases derived from 3,5-Bu(2)(t)-salicylaldehyde. AB - Nickel(II) complexes of a series of N,N'-polymethylenebis(3,5-Bu(2)(t) salicylaldimine) ligands containing 2,4-di-Bu(2)(t)-phenol arms, NiL(x), were synthesized and their spectroscopic and redox properties were examined. The UV vis, (1)H NMR spectroscopic and magnetic results indicate that complexes NiL(1) NiL(4) unlike NiL(5) and NiL(6) have a square-planar structure in the solid state and in solution. Cyclic voltammograms of NiL(x) (x=1-4) complexes displayed two step oxidation processes. The first oxidation peak potentials of all Ni(II) complexes corresponds to the reversible one-electron oxidation process of the metal center, yielding Ni(III) species. The second oxidation peak of the complexes was assigned as the ligand based oxidation generating a coordinated phenoxyl radical species. PMID- 15876550 TI - Biogeological Raman spectroscopic studies of Antarctic lacustrine sediments. AB - Analysis of lacustrine sediments is an accepted method for deciphering the palaeoenvironment of a lake's catchment area, as each strata of the sediment gives information about the rock type it was eroded from and also the state of the lake, i.e. oxic or anoxic. Antarctica has long been accepted as a putative analogue for Mars, so the analysis of Antarctic material may give results that can be compared to sediments on Mars. Raman spectroscopy has been selected as the method of analysis as it does not destroy the sample, can be used in situ and requires very little sample preparation. It is a suitable method for analysing both inorganic and organic matter and a miniature spectrometer is currently being developed for use in the field. The results from the spectrometers can serve as a guide for analysing sediments on Mars. It has been shown that Raman spectroscopy can detect and differentiate between oxic and anoxic sediments. Both 1064 and 785 nm wavelengths are suitable for laser excitation of organic and inorganic matter. PMID- 15876551 TI - Efficacy of ribavirin monotherapy following combination therapy with interferon alfa-2b and ribavirin for patients with chronic hepatitis C. AB - Reappearance of HCV-RNA followed by exacerbation of biochemical parameters after combination therapy consisting of interferon and ribavirin is an obstacle to achieve sustained response and improve long-term prognosis. We hypothesed that ribavirin monotherapy after 6 months of combination therapy may improve sustained viral and biochemical responses, and conducted a prospective, randomized and controlled study. Thirty-eight patients with chronic hepatitis C treated with combination therapy for 6 months and had no detectable serum HCV-RNA were enrolled, and allocated into two arms. Group I (n=19) was continuously administered oral ribavirin for additional 6 months, and group II (n=19) was followed up without any further treatment. At the end of trial, HCV-RNA negativity was 11/19 (58%) in group I, and 6/19 (32%) in group II (p=0.191). Multivariant analysis demonstrated that ribavirin monotherapy was not a predictor for the eradication of HCV-RNA. In cases without sustained viral responses, serum ALT levels at baseline and the end of 48 weeks' trial were 54.6 and 44.4 in group I (p=0.237), and significant reduction with ribavirin monotherapy was not observed. In conclusion, ribavirin monotherapy following combination therapy fails to improve sustained viral response as well as biochemical response. PMID- 15876552 TI - Macrophage inflammatory protein-1alpha plays a crucial role in concanavalin A induced liver injury through induction of proinflammatory cytokines in mice. AB - BACKGROUND/AIMS: : The chemokines play roles in the development of immune mediated liver diseases. In this study, we investigate the involvement of macrophage inflammatory protein-1alpha (MIP-1alpha), one of the CC chemokines in concanavalin A (Con A)-induced liver injury in mice. METHODS: : Liver injury was induced by intravenous injection of Con A. Anti-mouse MIP-1alpha antibody, recombinant murine-MIP-1alpha and gadolinium chloride (GdCl(3)) were administrated prior to Con A injection. Plasma alanine aminotransferase (ALT), MIP-1alpha, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) levels were determined and histological assessment of the liver was performed. RESULTS: : Plasma MIP-1alpha level was elevated after Con A injection. The elevated plasma ALT level, mortality rate and histological change after Con A injection were inhibited by anti-MIP-1alpha antibody pretreatment. The elevated plasma ALT level after Con A injection was further enhanced by recombinant murine MIP-1alpha. The elevated plasma TNF-alpha and IFN-gamma levels after Con A injection were inhibited by anti-MIP-1alpha antibody, and enhanced by recombinant murine-MIP-1alpha. GdCl(3) pretreatment inhibited the elevated plasma MIP-1alpha and ALT levels. CONCLUSIONS: : These findings suggest that MIP-1alpha is produced from Kupffer cells after Con A injection, and this CC chemokine plays a crucial role in Con A-induced liver injury through induction of proinflammatory cytokines. PMID- 15876553 TI - The effect of contraction intensity on motor unit number estimates of the tibialis anterior. AB - OBJECTIVE: To examine the effect of contractile level on motor unit number estimates (MUNEs) and establish the contraction intensity that will yield the most representative MUNE for the tibialis anterior (TA) muscle. METHODS: Surface and intramuscular electromyographic (EMG) signals were collected during a range of submaximal (threshold, 10, 20, 30 and 40% MVC) isometric dorsiflexion contractions using decomposition-enhanced spike-triggered averaging (DE-STA). Six MUNEs were calculated, one for each of the five intensities, and an ensemble sixth MUNE that had equal MU contributions from all intensities. RESULTS: Mean surface-motor unit potential sizes increased significantly (26-69 microV) and MUNEs decreased accordingly (226-91) as contraction intensity increased from threshold to 40% MVC, respectively (P<0.05). The ensemble MUNE was 153, and extrapolated to approximately 25% MVC. CONCLUSIONS: There was a significant and progressive decline in the MUNE as contraction intensity increased, confirming the importance of monitoring torque during data collection. The ensemble MUNE suggests that collecting EMG signals at a contraction intensity of approximately 25% MVC provides the most representative sample of the actual number and sizes of MUs in the TA. SIGNIFICANCE: Establishing appropriate contraction intensities improves the utility of DE-STA as a useful method for tracking changes to the MU pool in disease states and healthy aging. PMID- 15876554 TI - Nonconvulsive status epilepticus on treatment with levetiracetam. AB - In several studies the efficacy and tolerability of levetiracetam (LEV) have been demonstrated. We report two patients who developed nonconvulsive status epilepticus on treatment with LEV. Both patients never experienced status epilepticus before. One patient had a symptomatic epilepsy with complex partial seizures following radiotherapy of astrocytoma in 1985; the second patient had complex partial seizures due to mesial temporal sclerosis. Both patients received LEV 2000 mg/day. We postulate a correlation between occurrence of nonconvulsive status and treatment with LEV. This has not been described before apart from a single report of mentally retarded patients with status epilepticus on high dosages of LEV. PMID- 15876555 TI - Michael Trimble: the quintessential clinician scientist. PMID- 15876556 TI - What is effective treatment of depression in people with epilepsy? PMID- 15876557 TI - Neonatal maturation of the hypercapnic ventilatory response and central neural CO2 chemosensitivity. AB - The ventilatory response to CO2 changes as a function of neonatal development. In rats, a ventilatory response to CO2 is present in the first 5 days of life, but this ventilatory response to CO2 wanes and reaches its lowest point around postnatal day 8. Subsequently, the ventilatory response to CO2 rises towards adult levels. Similar patterns in the ventilatory response to CO2 are seen in some other species, although some animals do not exhibit all of these phases. Different developmental patterns of the ventilatory response to CO2 may be related to the state of development of the animal at birth. The triphasic pattern of responsiveness (early decline, a nadir, and subsequent achievement of adult levels of responsiveness) may arise from the development of several processes, including central neural mechanisms, gas exchange, the neuromuscular junction, respiratory muscles and respiratory mechanics. We only discuss central neural mechanisms here, including altered CO2 sensitivity of neurons among the various sites of central CO2 chemosensitivity, changes in astrocytic function during development, the maturation of electrical and chemical synaptic mechanisms (both inhibitory and excitatory mechanisms) or changes in the integration of chemosensory information originating from peripheral and multiple central CO2 chemosensory sites. Among these central processes, the maturation of synaptic mechanisms seems most important and the relative maturation of synaptic processes may also determine how plastic the response to CO2 is at any particular age. PMID- 15876558 TI - Postnatal development of ventilatory and arousal responses to hypoxia in human infants. AB - During the first year of life there is significant maturation of the hypoxic ventilatory response (HVR) in human infants. Compared with adults, healthy term infants have an immature HVR until at least 6 months of age. There are few studies in infants on the effects of sleep state on the HVR but these suggest that at early postnatal ages there is initially no sleep-state related difference; this is followed by a developmental trend towards the adult situation in which the response is depressed in REM sleep compared with NREM. Maternal cigarette smoking is a major risk factor for SIDS and the mechanism for this may involve a depressed HVR in the exposed infant; however studies are limited and the wide variation in cigarette consumption makes interpretation of results difficult. Arousal responses to hypoxia are of vital importance and a failure to arouse has been implicated in SIDS. Sleeping infants frequently fail to arouse in response to hypoxia in QS, whereas in AS they invariably arouse; furthermore arousal latency is longer in QS compared with AS. The oxygen saturation at which infants arouse is not different between sleep states, suggesting that desaturation is more rapid in AS. In QS younger infants arouse more readily than at older ages and arousal is depressed by maternal smoking. These findings suggest that depression of the arousal response to hypoxia in AS may have life threatening consequences. Infants at increased risk for SIDS have been shown to have both depressed ventilatory and arousal responses to hypoxia, thus they may be at even greater risk. PMID- 15876559 TI - Effect of surfactant deficiency and surfactant replacement on airway patency in the piglet lung. AB - We investigated the effect of surfactant deficiency on airway patency and the effectiveness of surfactant replacement as either an instilled liquid bolus, a non-hygroscopic aerosol or a hygroscopic aerosol. Small airway patency was assessed in isolated piglet lungs by passing a continuous flow of gas though a cannulated airway. Occlusion was assessed by measuring increases in pressure in the cannula that resulted from airway obstruction. In surfactant-deficient conditions the amount of airway closure increased approximately three-fold. However, administration of exogenous surfactant as an instilled liquid bolus, non hygroscopic aerosol or a hygroscopic aerosol decreased airway closure such that it was statistically similar to that recorded prior to induction of surfactant deficiency, although the instilled and hygroscopic aerosol surfactant both appeared superior to the non-hygroscopic aerosol. These experiments showed that pulmonary surfactant does have a role in maintaining airway patency and that airway closure induced by surfactant deficiency could be reduced by administration of surfactant in any of the aforementioned forms. PMID- 15876560 TI - High performance liquid chromatographic determination of azithromycin in serum using fluorescence detection and its application in human pharmacokinetic studies. AB - A fast and sensitive high-performance liquid chromatographic method for determination of azithromycin in human serum using fluorescence detection was developed. The drug and an internal standard (clarithromycin) were extracted from serum using n-hexan and subjected to pre-column derivatization with 9 fluorenylmethyl chloroformate as labeling agent. Analysis was performed on a phenyl packing material column with sodium phosphate buffer containing 2 ml/l triethylamine (pH 5.9) and methanol (29:71, v/v) as the mobile phase. The standard curve was linear over the range of 10-500 ng/ml of azithromycin in human serum. The means between-days precision were from 13.3% (for 10 ng/ml) to 2% (500 ng/ml) and the within-day precision from 11.9 to 1.7% determined on spiked samples. The accuracy of the method was 100.7-107.2% (between days) and 100.3 107.8% (within day). The limit of quantification was 10 ng/ml. This method was applied in a bioequivalence study of four different azithromycin preparations in 12 healthy volunteers. PMID- 15876561 TI - Bone geometry and density in the skeleton of pre-pubertal gymnasts and school children. AB - We have studied the differences between the peripheral and axial skeleton of pre pubertal gymnasts and controls. We hypothesised that compared to controls, gymnasts would have larger and stronger radius and tibia diaphyses with greater bone mineral content and larger cross-sectional muscle area. At the distal metaphyseal sites of the radius and tibia, gymnasts would have greater bone cross sectional area and total and trabecular volumetric bone mineral density (vBMD). Differences between the lumbar spine, total body and body composition in gymnasts versus controls were also studied. Peripheral quantitative computed tomography (pQCT) was used to measure bone geometry, density and muscle of the peripheral skeleton; dual energy X-ray absorptiometry (DXA) for total body and axial measurements. Eighty-six pre-pubertal children, 44 gymnasts (mean age 9.0 years, range 5.4-11.9 years) and 42 controls (mean age 8.8 years, range 5.6-11.9 years) were studied. Eighty-four children were Caucasian, one child was mixed race, one Chinese. Data were adjusted for age, sex and height. Differences in the effect size between sexes were also tested. At the 50% radius diaphysis gymnasts had larger bones (9.2%, p = 0.0054) with greater cortical area (8.2%, p = 0.022) and stress strain index (surrogate measure of bone strength) than controls (13.6%, p = 0.015). The effect size was different between males and females for cortical thickness (p = 0.03). At the 65% tibia diaphysis, gymnasts had greater cortical area (5.3%, p = 0.057) and thickness (6.2%, p = 0.068) than controls; consequently, bone strength was 5.4% higher (p = 0.14). There were no significant differences in cortical volumetric bone mineral density (vBMD) at the radius or tibia diaphysis between the groups. There was a difference in effect size for tibia muscle cross-sectional area between the sexes (p = 0.035). At the distal radius and tibia total and trabecular vBMD was greater (Total: radius 17%, p < 0.0001, tibia: 5.7%, p = 0.0053; trabecular: radius 21%, p < 0.0001, tibia 4.5%, p = 0.11). Bone size was not different in gymnasts compared to controls Lumbar spine BMC (12.3%, p = 0.0007), areal bone mineral density (aBMD) (9.1%, p = 0.0006) and bone mineral apparent density (BMAD) (7.6%, p = 0.0047) were greater in gymnasts but vertebral size was not significantly different. Likewise, total body BMD (3.5%, p = 0.0057) and BMC (4.78%, p = 0.085) were greater in gymnasts but there were no differences in skeletal size. These data suggest site-specific differences in how the pre-pubertal skeleton develops in response to the repetitive loading it experiences when participating in regular gymnastics. At diaphyseal sites these differences are predominantly in the bone and muscle geometry and not density. Conversely, at trabecular sites, the differences are increased density rather than geometry. In conclusion, the present study has demonstrated skeletal differences between gymnasts and controls. These differences appear to be site and sex specific. PMID- 15876562 TI - Characterization of an archaeal malic enzyme from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. AB - Although the interconversion between C4 and C3 compounds has an important role in overall metabolism, limited information is available on the properties and regulation of enzymes acting on these metabolites in hyperthermophilic archaea. Malic enzyme is one of the enzymes involved in this interconversion, catalyzing the oxidative decarboxylation of malate to pyruvate as well as the reductive carboxylation coupled with NAD(P)H. This study focused on the enzymatic properties and expression profile of an uncharacterized homolog of malic enzyme identified in the genome of a heterotrophic, hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 (Tk-Mae). The amino acid sequence of Tk-Mae was 52-58% identical to those of malic enzymes from bacteria, whereas the similarities to the eukaryotic homologs were lower. Several catalytically important regions and residues were conserved in the primary structure of Tk-Mae. The recombinant protein, which formed a homodimer, exhibited thermostable malic enzyme activity with strict divalent cation dependency. The enzyme preferred NADP(+) rather than NAD(+), but did not catalyze the decarboxylation of oxaloacetate, unlike the usual NADP-dependent malic enzymes. The apparent Michaelis constant (K(m)) of Tk-Mae for malate (16.9 mM) was much larger than those of known enzymes, leading to no strong preference for the reaction direction. Transcription of the gene encoding Tk-Mae and intracellular malic enzyme activity in T. kodakaraensis were constitutively weak, regardless of the growth substrates. Possible roles of Tk-Mae are discussed based on these results and the metabolic pathways of T. kodakaraensis deduced from the genome sequence. PMID- 15876563 TI - Trace methane oxidation studied in several Euryarchaeota under diverse conditions. AB - We used (13)C-labeled methane to document the extent of trace methane oxidation by Archaeoglobus fulgidus, Archaeoglobus lithotrophicus, Archaeoglobus profundus, Methanobacterium thermoautotrophicum, Methanosarcina barkeri and Methanosarcina acetivorans. The results indicate trace methane oxidation during growth varied among different species and among methanogen cultures grown on different substrates. The extent of trace methane oxidation by Mb. thermoautotrophicum (0.05 +/- 0.04%, +/- 2 standard deviations of the methane produced during growth) was less than that by M. barkeri (0.15 +/- 0.04%), grown under similar conditions with H(2) and CO(2). Methanosarcina acetivorans oxidized more methane during growth on trimethylamine (0.36 +/- 0.05%) than during growth on methanol (0.07 +/ 0.03%). This may indicate that, in M. acetivorans, either a methyltransferase related to growth on trimethylamine plays a role in methane oxidation, or that methanol is an intermediate of methane oxidation. Addition of possible electron acceptors (O(2), NO(3) (-), SO(4) (2-), SO(3) (2-)) or H(2) to the headspace did not substantially enhance or diminish methane oxidation in M. acetivorans cultures. Separate growth experiments with FAD and NAD(+) showed that inclusion of these electron carriers also did not enhance methane oxidation. Our results suggest trace methane oxidized during methanogenesis cannot be coupled to the reduction of these electron acceptors in pure cultures, and that the mechanism by which methane is oxidized in methanogens is independent of H(2) concentration. In contrast to the methanogens, species of the sulfate-reducing genus Archaeoglobus did not significantly oxidize methane during growth (oxidizing 0.003 +/- 0.01% of the methane provided to A. fulgidus, 0.002 +/- 0.009% to A. lithotrophicus and 0.003 +/- 0.02% to A. profundus). Lack of observable methane oxidation in the three Archaeoglobus species examined may indicate that methyl-coenzyme M reductase, which is not present in this genus, is required for the anaerobic oxidation of methane, consistent with the "reverse methanogenesis" hypothesis. PMID- 15876564 TI - Active site of Zn(2+)-dependent sn-glycerol-1-phosphate dehydrogenase from Aeropyrum pernix K1. AB - The enzyme sn-glycerol-1-phosphate dehydrogenase (Gro1PDH, EC 1.1.1.261) is key to the formation of the enantiomeric configuration of the glycerophosphate backbone (sn-glycerol-1-phosphate) of archaeal ether lipids. This enzyme catalyzes the reversible conversion between dihydroxyacetone phosphate and glycerol-1-phosphate. To date, no information about the active site and catalytic mechanism of this enzyme has been reported. Using the sequence and structural information for glycerol dehydrogenase, we constructed six mutants (D144N, D144A, D191N, H271A, H287A and D191N/H271A) of Gro1PDH from Aeropyrum pernix K1 and examined their characteristics to clarify the active site of this enzyme. The enzyme was found to be a zinc-dependent metalloenzyme, containing one zinc ion for every monomer protein that was essential for activity. Site-directed mutagenesis of D144 increased the activity of the enzyme. Mutants D144N and D144A exhibited low affinity for the substrates and higher activity than the wild type, but their affinity for the zinc ion was the same as that of the wild type. Mutants D191N, H271A and H287A had a low affinity for the zinc ion and a low activity compared with the wild type. The double mutation, D191N/H271A, had no enzyme activity and bound no zinc. From these results, it was clarified that residues D191, H271 and H287 participate in the catalytic activity of the enzyme by binding the zinc ion, and that D144 has an effect on substrate binding. The structure of the active site of Gro1PDH from A. pernix K1 seems to be similar to that of glycerol dehydrogenase, despite the differences in substrate specificity and biological role. PMID- 15876565 TI - Novel RepA-MCM proteins encoded in plasmids pTAU4, pORA1 and pTIK4 from Sulfolobus neozealandicus. AB - Three plasmids isolated from the crenarchaeal thermoacidophile Sulfolobus neozealandicus were characterized. Plasmids pTAU4 (7,192 bp), pORA1 (9,689 bp) and pTIK4 (13,638 bp) show unusual properties that distinguish them from previously characterized cryptic plasmids of the genus Sulfolobus. Plasmids pORA1 and pTIK4 encode RepA proteins, only the former of which carries the novel polymerase-primase domain of other known Sulfolobus plasmids. Plasmid pTAU4 encodes a mini-chromosome maintenance protein homolog and no RepA protein; the implications for DNA replication are considered. Plasmid pORA1 is the first Sulfolobus plasmid to be characterized that does not encode the otherwise highly conserved DNA-binding PlrA protein. Another encoded protein appears to be specific for the New Zealand plasmids. The three plasmids should provide useful model systems for functional studies of these important crenarchaeal proteins. PMID- 15876566 TI - Recombinant production of Zymomonas mobilis pyruvate decarboxylase in the haloarchaeon Haloferax volcanii. AB - The unusual physiological properties of archaea (e.g., growth in extreme salt concentration, temperature and pH) make them ideal platforms for metabolic engineering. Towards the ultimate goal of modifying an archaeon to produce bioethanol or other useful products, the pyruvate decarboxylase gene of Zymomonas mobilis (Zm pdc) was expressed in Haloferax volcanii. This gene has been used successfully to channel pyruvate to ethanol in various Gram-negative bacteria, including Escherichia coli. Although the ionic strength of the H. volcanii cytosol differs over 15-fold from that of E. coli, gel filtration and circular dichroism revealed no difference in secondary structure between the ZmPDC protein isolated from either of these hosts. Like the E. coli purified enzyme, ZmPDC from H. volcanii catalyzed the nonoxidative decarboxylation of pyruvate. A decrease in the amount of soluble ZmPDC protein was detected as H. volcanii transitioned from log phase to late stationary phase that was inversely proportional to the amount of pdc-specific mRNA. Based on these results, proteins from non-halophilic organisms can be actively synthesized in haloarchaea; however, post transcriptional mechanisms present in stationary phase appear to limit the amount of recombinant protein expressed. PMID- 15876567 TI - Identification of replication origins in archaeal genomes based on the Z-curve method. AB - The Z-curve is a three-dimensional curve that constitutes a unique representation of a DNA sequence, i.e., both the Z-curve and the given DNA sequence can be uniquely reconstructed from the other. We employed Z-curve analysis to identify one replication origin in the Methanocaldococcus jannaschii genome, two replication origins in the Halobacterium species NRC-1 genome and one replication origin in the Methanosarcina mazei genome. One of the predicted replication origins of Halobacterium species NRC-1 is the same as a replication origin later identified by in vivo experiments. The Z-curve analysis of the Sulfolobus solfataricus P2 genome suggested the existence of three replication origins, which is also consistent with later experimental results. This review aims to summarize applications of the Z-curve in identifying replication origins of archaeal genomes, and to provide clues about the locations of as yet unidentified replication origins of the Aeropyrum pernix K1, Methanococcus maripaludis S2, Picrophilus torridus DSM 9790 and Pyrobaculum aerophilum str. IM2 genomes. PMID- 15876568 TI - Biosynthesis of ribose-5-phosphate and erythrose-4-phosphate in archaea: a phylogenetic analysis of archaeal genomes. AB - A phylogenetic analysis of the genes encoding enzymes in the pentose phosphate pathway (PPP), the ribulose monophosphate (RuMP) pathway, and the chorismate pathway of aromatic amino acid biosynthesis, employing data from 13 complete archaeal genomes, provides a potential explanation for the enigmatic phylogenetic patterns of the PPP genes in archaea. Genomic and biochemical evidence suggests that three archaeal species (Methanocaldococcus jannaschii, Thermoplasma acidophilum and Thermoplasma volcanium) produce ribose-5-phosphate via the nonoxidative PPP (NOPPP), whereas nine species apparently lack an NOPPP but may employ a reverse RuMP pathway for pentose synthesis. One species (Halobacterium sp. NRC-1) lacks both the NOPPP and the RuMP pathway but may possess a modified oxidative PPP (OPPP), the details of which are not yet known. The presence of transketolase in several archaeal species that are missing the other two NOPPP genes can be explained by the existence of differing requirements for erythrose-4 phosphate (E4P) among archaea: six species use transketolase to make E4P as a precursor to aromatic amino acids, six species apparently have an alternate biosynthetic pathway and may not require the ability to make E4P, and one species (Pyrococcus horikoshii) probably does not synthesize aromatic amino acids at all. PMID- 15876569 TI - Higher-level classification of the Archaea: evolution of methanogenesis and methanogens. AB - We used a phylogenetic approach to analyze the evolution of methanogenesis and methanogens. We show that 23 vertically transmitted ribosomal proteins do not support the monophyly of methanogens, and propose instead that there are two distantly related groups of extant archaea that produce methane, which we have named Class I and Class II. Based on this finding, we subsequently investigated the uniqueness of the origin of methanogenesis by studying both the enzymes of methanogenesis and the proteins that synthesize its specific coenzymes. We conclude that hydrogenotrophic methanogenesis appeared only once during evolution. Genes involved in the seven central steps of the methanogenic reduction of carbon dioxide (CO(2)) are ubiquitous in methanogens and share a common history. This suggests that, although extant methanogens produce methane from various substrates (CO(2), formate, acetate, methylated C-1 compounds), these archaea have a core of conserved enzymes that have undergone little evolutionary change. Furthermore, this core of methanogenesis enzymes seems to originate (as a whole) from the last ancestor of all methanogens and does not appear to have been horizontally transmitted to other organisms or between members of Class I and Class II. The observation of a unique and ancestral form of methanogenesis suggests that it was preserved in two independent lineages, with some instances of specialization or added metabolic flexibility. It was likely lost in the Halobacteriales, Thermoplasmatales and Archaeoglobales. Given that fossil evidence for methanogenesis dates back 2.8 billion years, a unique origin of this process makes the methanogenic archaea a very ancient taxon. PMID- 15876570 TI - The selective cyclooxygenase-2 inhibitor SC-236 reduces liver fibrosis by mechanisms involving non-parenchymal cell apoptosis and PPARgamma activation. AB - The importance of inflammation in initiating the sequence of events that lead to liver fibrosis is increasingly recognized. In this study, we tested the effects of SC-236, a selective cyclooxygenase (COX)-2 inhibitor, in rats with carbon tetrachloride (CCl4)-induced liver fibrosis. Livers from CCl4-treated rats showed increased COX-2 expression and 15-deoxy-prostaglandin (PG)J2 (15d-PGJ2) formation, as well as decreased peroxisome proliferator-activated receptor (PPAR)gamma expression. In these animals, SC-236 reduced liver fibrosis as revealed by histological analysis and by a reduction in hepatic hydroxyproline levels, metalloproteinase-2 activity, and alpha-smooth muscle actin expression. Interestingly, SC-236 normalized 15d-PGJ2 levels and restored PPARgamma expression in the liver of CCl4-treated rats. In isolated hepatic stellate cells (HSCs)--the major player in liver fibrogenesis--and Kupffer cells--the cell type primarily responsible for increased hepatic COX-2-SC-236 exhibited remarkable pro apoptotic and growth inhibitory properties. Of interest, SC-236 decreased HSC viability to a similar extent than the PPARgamma ligand rosiglitazone. Moreover, SC-236 significantly induced PPARgamma expression in HSCs and acted as a potent PPARgamma agonist in a luciferase-reporter trans-activation assay. These data indicate that, by mechanisms involving non-parenchymal cell apoptosis and PPARgamma activation, the selective COX-2 inhibitor SC-236 might have therapeutic potential for prevention of liver fibrosis. PMID- 15876571 TI - The increased diagnosis of "juvenile bipolar disorder": what are we treating? PMID- 15876572 TI - False positivity for Aspergillus antigenemia with amoxicillin-clavulonic acid. PMID- 15876573 TI - The cellular perspective on mammary gland development: stem/progenitor cells and beyond. AB - Study of the mammary gland at the stem cell level is necessary for understanding mammary gland development. Knowledge of mammary gland development and growth is the first step toward formulating strategies to improve milk production. The success of these strategies requires an understanding of the dynamics of adult stem cells and their progeny in the development of the bovine mammary gland. The stem cell lineage pathway begins with adult stem cells and ends with the production of terminally differentiated cells. The progression of adult stem cells along the mammary gland stem cell lineage pathway requires the coordination of many events. One important event in this process is cell differentiation. This differentiation process evolves with a gradient appearance of cell organelles progressing from stem cells to terminally differentiated cells. To dissect differentiation, mechanisms that regulate stem cells to differentiate toward a particular cell fate must be identified. Ultrastructural characteristics assist in distinguishing cells in various stages of differentiation in the mammary gland cell lineage pathway. Cells in the lineage pathway can become either epithelial cells or myoepithelial cells. Epithelial cells function in the production and secretion of milk, whereas myoepithelial cells assist epithelial cells in milk secretion. This review focuses on current concepts regarding adult stem cells and the recent progress on bovine mammary gland stem/progenitor cell development and differentiation. Multistep strategies that incorporate manipulation of the mechanisms influencing lineage choices in the mammary gland will produce beneficial effects on mammary gland development and milk production. PMID- 15876574 TI - Impacts of the source and amount of crude protein on the intestinal supply of nitrogen fractions and performance of dairy cows. AB - The objective of this article was to review and summarize the significance of the amount and source of dietary crude protein supplements on the supply of nitrogen fractions passing to the small intestine and the performance of lactating dairy cows. A meta-analysis was used to evaluate 2 data sets, one for nitrogen flow to the small intestine and one for performance of cows. The response of dairy cows to rumen-undegradable protein supplements is variable. A portion of this variable response from research trials is explained by the source of crude protein in the control diet, the proportion and source of rumen-undegradable protein in the experimental diet, the effect of rumen-undegradable protein on microbial protein outflow from the rumen, the degradability and amino acid content of the rumen undegradable protein, and the crude protein percentage of the diet. Compared with soybean meal, the mean milk production responses to feeding rumen-undegradable protein supplements ranged from -2.5 to +2.75%. Because of the large variation and small magnitude of response when rumen-undegradable protein supplements are fed compared with soybean meal, efficiency of nitrogen utilization and the cost to benefit ratio for these crude protein supplements may determine the source and amount of crude protein to feed to dairy cows in the future. PMID- 15876575 TI - Nitrogen metabolism in the rumen. AB - Protein metabolism in the rumen is the result of metabolic activity of ruminal microorganisms. The structure of the protein is a key factor in determining its susceptibility to microbial proteases and, thus, its degradability. Ruminal protein degradation is affected by pH and the predominant species of microbial population. Ruminal proteolytic activity decreases as pH decreases with high forage dairy cattle-type rations, but not in high-concentrate beef-type rations. Accumulation of amino acid (AA) N after feeding suggests that AA uptake by rumen microorganisms could be the limiting factor of protein degradation in the rumen. In addition, there are several AA, such as Phe, Leu, and Ile, that are synthesized by rumen microorganisms with greater difficulty than other AA. The most common assessment of efficiency of microbial protein synthesis (EMPS) is determination of grams of microbial N per unit of rumen available energy, typically expressed as true organic matter or carbohydrates fermented. However, EMPS is unable to estimate the efficiency at which bacteria capture available N in the rumen. An alternative and complementary measure of microbial protein synthesis is the efficiency of N use (ENU). In contrast to EMPS, ENU is a good measurement for describing efficiency of N capture by ruminal microbes. Using EMPS and ENU, it was concluded that optimum bacterial growth in the rumen occurs when EMPS is 29 g of bacterial N/kg of fermented organic matter, and ENU is 69%, implying that bacteria would require about 1.31 x rumen-available N per unit of bacterial N. Because the distribution of N within bacterial cells changes with rate of fermentation, AA N, rather than total bacterial N should be used to express microbial protein synthesis. PMID- 15876577 TI - A family tree in every gene. PMID- 15876578 TI - Strategic analysis in evolutionary genetics and the theory of games. AB - This paper is written in memory of John Maynard Smith. In a brief survey it discusses essential aspects of how game theory in biology relates to its counterpart in economics, the major transition in game theory initiated by Maynard Smith, the discrepancies between genetic and phenotypic models in evolutionary biology, and a balanced way of reconciling these models. In addition, the paper discusses modern problems in understanding games at the genetic level using the examples of conflict between endosymbionts and their hosts, and the molecular interactions between parasites and the mammalian immune system. PMID- 15876579 TI - Inbreeding and sex: canalization, plasticity and sexual selection. PMID- 15876580 TI - Fertility, mating behaviour and sexual selection in Drosophila subobscura. J. Genet. 1956, 54, 261-279. PMID- 15876581 TI - Acclimatization to high temperatures in inbred and outbred Drosophila subobscura. J. Genet. 1956, 54, 497-505. PMID- 15876582 TI - Coincident natural selection of CCR5Delta32 and C282Y in Europe: to be or not to be? PMID- 15876583 TI - Exact tandem repeats analyzer (E-TRA): a new program for DNA sequence mining. AB - Exact Tandem Repeats Analyzer 1.0 (E-TRA) combines sequence motif searches with keywords such as 'organs', 'tissues', 'cell lines' and 'development stages' for finding simple exact tandem repeats as well as non-simple repeats. E-TRA has several advanced repeat search parameters/options compared to other repeat finder programs as it not only accepts GenBank, FASTA and expressed sequence tags (EST) sequence files, but also does analysis of multiple files with multiple sequences. The minimum and maximum tandem repeat motif lengths that E-TRA finds vary from one to one thousand. Advanced user defined parameters/options let the researchers use different minimum motif repeats search criteria for varying motif lengths simultaneously. One of the most interesting features of genomes is the presence of relatively short tandem repeats (TRs). These repeated DNA sequences are found in both prokaryotes and eukaryotes, distributed almost at random throughout the genome. Some of the tandem repeats play important roles in the regulation of gene expression whereas others do not have any known biological function as yet. Nevertheless, they have proven to be very beneficial in DNA profiling and genetic linkage analysis studies. To demonstrate the use of E-TRA, we used 5,465,605 human EST sequences derived from 18,814,550 GenBank EST sequences. Our results indicated that 12.44% (679,800) of the human EST sequences contained simple and non-simple repeat string patterns varying from one to 126 nucleotides in length. The results also revealed that human organs, tissues, cell lines and different developmental stages differed in number of repeats as well as repeat composition, indicating that the distribution of expressed tandem repeats among tissues or organs are not random, thus differing from the un-transcribed repeats found in genomes. PMID- 15876584 TI - Comparative studies on codon usage pattern of chloroplasts and their host nuclear genes in four plant species. AB - A detailed comparison was made of codon usage of chloroplast genes with their host (nuclear) genes in the four angiosperm species Oryza sativa, Zea mays, Triticum aestivum and Arabidopsis thaliana. The average GC content of the entire genes, and at the three codon positions individually, was higher in nuclear than in chloroplast genes, suggesting different genomic organization and mutation pressures in nuclear and chloroplast genes. The results of Nc-plots and neutrality plots suggested that nucleotide compositional constraint had a large contribution to codon usage bias of nuclear genes in O. sativa, Z. mays, and T. aestivum, whereas natural selection was likely to be playing a large role in codon usage bias in chloroplast genomes. Correspondence analysis and chi-test showed that regardless of the genomic environment (species) of the host, the codon usage pattern of chloroplast genes differed from nuclear genes of their host species by their AU-richness. All the chloroplast genomes have predominantly A- and/or U-ending codons, whereas nuclear genomes have G-, C- or U-ending codons as their optimal codons. These findings suggest that the chloroplast genome might display particular characteristics of codon usage that are different from its host nuclear genome. However, one feature common to both chloroplast and nuclear genomes in this study was that pyrimidines were found more frequently than purines at the synonymous codon position of optimal codons. PMID- 15876585 TI - Mutational analysis of the BRCA1 gene in 30 Czech ovarian cancer patients. AB - Ovarian cancer is one of the most severe of oncological diseases. Inherited mutations in cancer susceptibility genes play a causal role in 5-10% of newly diagnosed tumours. BRCA1 and BRCA2 gene alterations are found in the majority of these cases. The aim of this study was to analyse the BRCA1 gene in the ovarian cancer risk group to characterize the spectrum of its mutations in the Czech Republic. Five overlapping fragments amplified on both genomic DNA and cDNA were used to screen for the whole protein-coding sequence of the BRCA1 gene. These fragments were analysed by the protein truncation test (PTT) and direct sequencing. Three inactivating mutations were identified in the group of 30 Czech ovarian cancer patients: the 5382insC mutation in two unrelated patients and a deletion of exons 21 and 22 in another patient. In addition, we have found an alternatively spliced product lacking exon 5 in two other unrelated patients. The 5382insC is the most frequent alteration of the BRCA1 gene in Central and Eastern Europe. The deletion of exons 21 and 22 affects the BRCT functional domain of the BRCA1 protein. Although large genomic rearragements are known to be relatively frequent in Western European populations, no analyses have been performed in our region yet. PMID- 15876586 TI - JP-3 gene polymorphism in a healthy population of Serbia and Montenegro. AB - Expansions of CTG repeats in JP-3 gene are associated with a phenotype similar to Huntington disease. These expansions are the cause of Huntington disease like-2 (HDL-2) phenotype. CTG repeats in JP-3 gene are polymorphic in healthy population. Analyses of CTG repeat polymorphism of JP-3 gene in various healthy populations could help in estimating the population at risk for developing HDL-2. CTG repeat polymorphism of JP-3 gene was analysed in healthy population of Serbia and Montenegro. Study included 198 unrelated subjects. Analyses of JP-3 locus were performed using PCR and sequencing. Six different JP-3 alleles were obtained and they were in the range of 11 to 18 CTG repeats showing a bimodal distribution, with peaks at 14 and 16. Results show that the distribution of JP-3 alleles in population of Serbia and Montenegro is consistent with distributions in other analysed populations. The absence of alleles with more then 18 CTG repeats suggests that HDL-2 is very rare in the populations of Serbia and Montenegro. PMID- 15876587 TI - Chromosomal localization of autosomal mutations in Drosophila nasuta nasuta and Drosophila nasuta albomicans. PMID- 15876588 TI - Genetic diversity analysis of five cattle breeds native to China using microsatellites. PMID- 15876589 TI - Orchid flower evolution. PMID- 15876591 TI - Ernst Mayr (1904-2005). PMID- 15876592 TI - Specific learning disability: the invisible handicap. PMID- 15876593 TI - Long-term outcome in children with primary distal renal tubular acidosis. AB - OBJECTIVE: To evaluate complications in adequately treated children with distal renal tubular acidosis (RTA) and to identify factors influencing their development. DESIGN: Records of patients with primary distal RTA followed for 2 or more years at this hospital were reviewed. Case records were examined for age at onset of symptoms and at initiation of treatment, treatment details, follow-up and complications. Height, weight and growth velocity were expressed as standard deviation score (SDS) during different periods of follow-up. Regression analysis was performed to evaluate factors influencing increase in height and weight SDS. P value of less than 0.05 was considered significant. RESULTS: Of 18 patients (eleven boys), the diagnosis was established at the median (range) age of 6 yr (1.5-13 yr). These patients were followed up for a median (range) period of 4 yr (2-18.5 yr). Short stature (height SDS <-2) was noted in all patients at the time of diagnosis with median (range) height SDS of -5.2(-7.5 - -0.4). All patients had failure to thrive with median (range) weight SDS of -3(-5.7 - -1.5). Height SDS increased by median (range) of 2 (1.2-5.5) to become -2.7(-4.8 - -1.1) at last follow-up. Weight SDS increased by median (range) of 0.9 (-0.6 - 2.8) to become -2.4 (-4 - -0.5). Median (range) growth velocity SDS decreased from 3 (1 16) during first year of treatment to 1(-0.3 - 7) at four years with an increase in mean height SDS by 1.3 during the first two years of treatment. Height SDS at last follow-up was not influenced by the age at initiation of treatment, follow up duration, initial height SDS or severity of acidosis at diagnosis. Increase in height SDS correlated negatively with base excess and height SDS at diagnosis, and positively with follow-up duration on univariate analyses. Initial height SDS was the only factor that influenced increase in height SDS on multivariate analyses. Increase in weight SDS was negatively correlated with base excess and initial weight SDS with significant association with initial weight SDS, on multivariate analyses. All patients had rickets at the time of diagnosis with pathological fractures in four. Rickets resolved without treatment with vitamin D in seventeen patients after a median (range) period of 4 mo (1-12 mo). Eight patients (44.4%) had nephrocalcinosis at diagnosis. Twenty-four hour urine calcium excretion was higher in those with nephrocalcinosis (p = 0.01). Creatinine clearance remained normal in all except one who progressed to renal failure. CONCLUSION: There is a need for early diagnosis, appropriate treatment and regular follow-up of patients with distal RTA for improving outcome. PMID- 15876594 TI - Nitric oxide metabolites in induced sputum: a noninvasive marker of airway inflammation in asthma. AB - OBJECTIVE: This study was designed to examine for nitric oxide (NO) metabolites in induced sputum as a marker of airway inflammation in asthmatic children. DESIGN. Prospective interventional SETTING: Pediatric Allergy and Asthma Clinic of a tertiary care referral hospital in Northern India. SUBJECTS: Twenty-one children with asthma who were not receiving corticosteroids for the preceding 3 months and 10 healthy controls were enrolled. METHODS: Hypertonic saline-induced sputum was obtained at study entry in controls, and at study entry and after 6 weeks of inhaled corticosteroid (ICS) therapy in asthmatic children. Fresh expectorated sputum was treated with dithiothreitol and cytospinned for cell count. NO metabolites were measured in the supernatant by the modified Griess reaction. RESULTS: Asthmatic children, compared with controls, had significantly higher concentration of NO metabolites (22.4 +/- 209.69 vs 39.2 +/- 15.9 (moL/L, P <0.01) and a higher percentage of eosinophils (15.3 +/- 12.0 vs 0.8 +/- 1.1%, P <0.01) in induced sputum. Both NO metabolites and eosinophil percentage declined following treatment with ICS for 6 weeks (P <0.01). CONCLUSION: The study confirms that the level of NO metabolites is increased in the tracheobronchial secretions of asthmatic children and decreases following ICS therapy. Measurement of NO metabolites in induced sputum may be useful for monitoring airway inflammation in children with asthma. PMID- 15876595 TI - Molecular aspects of Down syndrome. AB - Molecular aspects of Down syndrome (DS), a major genetic cause for mental retardation, commonly associated with trisomy 21 are discussed. Two different hypotheses have been speculated to better understand the disease. One believes that increased gene dosage contributes to the phenotypic abnormalities; the other correlates genetic imbalance with DS pathogenesis. To sustain these hypotheses, different murine models have been developed. Experimental models as well as sequencing of human chromosome 21 helped in speculating a few possible candidate genes for DS. However, the phenotypic changes involved with this neurological disorder vis-a-vis the enhanced number of genes, still remain unexplained. Improvement in screening pattern, model system, as well as better understanding of the disease etiology may help in developing efficacious therapeutic regimes for DS. PMID- 15876596 TI - Dietary intake and growth pattern of children 9-36 months of age in an urban slum in Delhi. AB - This investigation deals with the dietary intakes and growth in early childhood in poor communities. Five hundred and forty five children, 9-36 months of age, and their caretakers (mothers) were selected randomly and invited to participate in the study. The food and nutrient intake of children was assessed using a 10 item food frequency and amount questionnaire (FAQ). The anthropometric nutritional status was assessed by the indices height-for-age (H/A), weight-for age (W/A), weight-for-height (Wt/Ht) and MUAC, according to Indian and NCHS standards. The results showed that the intake of cereals, pulses, roots, green leafy vegetables (GLVs), other vegetables, fruits, sugar, fats and oils among children was grossly inadequate. The nutrient intake for energy was 56% of the current RDA. Anthropometric analysis revealed that the children were grossly undernourished. Seventy five per cent children were underweight (<-2 SD), while 35% severely undernourished (<-3 SD). Approximately, 74% children were having short stature (chronic malnutrition) with 39% severely stunted. Nineteen per cent children were excessively thin (wasted). The data regarding the degree of malnutrition among children demonstrated that 9.6% girls in 9-36 months of age had severe malnutrition as compared to 6.5% males. The maximum prevalence of severe malnutrition was in the age 31-36 months (10%) followed by 9.6% in 13-18 months. The moderate degree of malnutrition was around 30 to 33% in age group 13 36 months. The evidence from the study provides a strong basis to suggest low food intake as the main cause of under/malnutrition and growth retardation (stunting) in early childhood in poor communities. PMID- 15876597 TI - Hemoglobin E-beta thalassemia: factors affecting phenotype. AB - The phenotype of E-beta-thalassemia is affected by several genetic factors. The aim of this study was to analyze severity of E-beta-thalassemia and correlate with HbE, HbF, E/F ratios, beta-mutation and Xmn I polymorphism. Thirty cases of E-beta-thalassemia (23 with childhood onset) were studied. HbE levels were quantitated by HPLC. Xmn1 polymorphism and beta-mutations were studied by PCR RFLP and ARMS respectively. Commonest features were pallor (100%), splenomegaly (74%), and hepatomegaly (65%), 43% (10/23) were on regular transfusions at diagnosis. One case presented with paraplegia. Patients heterozygous for Xmn I polymorphism (+/-) had later onset (>3 yrs) compared to homozygous (-/-) absence (0.5-2.8 yrs). Most (69.6%) showed beta-mutation IVS 1-5 (G-->C). Negative correlation was found between age of onset and HbE. Thus, presentation is similar to previously reported Thai cases. Heterozygosity of Xmn I polymorphism also delays disease onset. Early diagnosis facilitates appropriate management and prenatal diagnosis. PMID- 15876598 TI - Avoidance of food allergens in childhood asthma. AB - Twenty-four patients of moderate persistent perennial asthma with documented aggravation to severe persistent asthma, during monsoon season in the past two years, were put on specific elimination diet during August and September. The diet was based on results of in-vitro allergy tests for a selected food panel. On specific elimination diet, five patients improved to mild persistent asthma and twelve patients improved to mild persistent asthma with occasional exacerbations . Six patients remained at moderate persistent asthma and only one patient deteriorated to severe persistent asthma. These results indicate that food avoidance may help in asthma control in children. PMID- 15876599 TI - Acute disseminated encephalomyelitis--a case series. AB - Acute disseminated encephalomyelitis(ADEM) is an uncommon inflammatory demyelinating disease involving central nervous system white matter. A series of seven cases seen over a period of one year have been described. The patients presented with acute onset of focal neurological deficit (3 patients), alteration of sensorium(3) and ataxia(1). CT scan showed non specific hypodensity in 4 cases and was normal in 3 patients . MRI showed characteristic radiological changes. Complete recovery was seen in 3 patients , one died and 3 were left with sequelae. PMID- 15876600 TI - Yunis-Varon syndrome. AB - Yunis-Varon syndrome is a rare, autosomal recessive syndrome characterized by growth retardation, defective growth of the cranial bones along with complete or partial absence of the clavicles (cleidocranial dysplasia), characteristic facial features, and/or abnormalities of the fingers and/or toes. PMID- 15876601 TI - Synchronous germinomas in the pineal and suprasellar region. AB - Synchronous primary intracranial germ cell tumors are rare. Only 5-10% of all germ cell tumors are found as synchronous lesion in pineal and suprasellar region. They are also known by the entity "double mid-line atypical teratoma". An 11-year-old male child presented with polyuria, polydipsia and features of raised intracranial tension. CT scan head revealed well-defined homogenously enhancing lesions in the pineal and suprasellar region. Histopathology examination showed the lesion to be of germ cell origin. PMID- 15876602 TI - Multiple hypoechoic lesions in spleen and Mycoplasma pneumoniae infection. AB - An 8-year-old boy was admitted because of recurrent fever for 1 month with increased CRP and ESR. Ultrasound reviewed multiple, small, hypo-echoic, rounded and wedge-shaped nodules with diffuse blood flow in spleen and enlarged abdominal lymph nodes. The spleen was enlarged and no echoic space was found in the largest lesion on 5th day. After a positive mycoplasma pneumoniae (MP) IgM was reported on 6th day, azithromycin was used intravenously. The temperature returned to normal and CRP and ESR improved in a short period. The lesions and lymphadenopathy disappeared and MP IgM antibody became negative 6 months later. PMID- 15876603 TI - Lichen simplex. PMID- 15876604 TI - Short rib polydactyly syndrome type I (Saldino-Noonan syndrome). PMID- 15876605 TI - Solitary mastocytoma. PMID- 15876606 TI - Intestinal strongyloidiasis and common variable immunodeficiency syndrome. PMID- 15876607 TI - Esophageal foreign body mimicking esophageal atresia. PMID- 15876608 TI - Continuous positive airway pressure-a gentler approach to ventilation. PMID- 15876609 TI - Giant omental cyst masquerading as hemorrhagic ascites. PMID- 15876610 TI - Betamethasone abuse in infancy. PMID- 15876611 TI - Cardiac murmur in neonates. PMID- 15876612 TI - Empyema following kerosene poisoning. PMID- 15876613 TI - Gender differences in nutritional status. PMID- 15876614 TI - Immunization status of children in Goa. PMID- 15876615 TI - A bedside dipstick method to detect Plasmodium falciparum. PMID- 15876616 TI - An efficient rhythmic component expression and weighting synthesis strategy for classifying motor imagery EEG in a brain-computer interface. AB - The recognition of mental states during motor imagery tasks is crucial for EEG based brain-computer interface research. We have developed a new algorithm by means of frequency decomposition and weighting synthesis strategy for recognizing imagined right- and left-hand movements. A frequency range from 5 to 25 Hz was divided into 20 band bins for each trial, and the corresponding envelopes of filtered EEG signals for each trial were extracted as a measure of instantaneous power at each frequency band. The dimensionality of the feature space was reduced from 200 (corresponding to 2 s) to 3 by down-sampling of envelopes of the feature signals, and subsequently applying principal component analysis. The linear discriminate analysis algorithm was then used to classify the features, due to its generalization capability. Each frequency band bin was weighted by a function determined according to the classification accuracy during the training process. The present classification algorithm was applied to a dataset of nine human subjects, and achieved a success rate of classification of 90% in training and 77% in testing. The present promising results suggest that the present classification algorithm can be used in initiating a general-purpose mental state recognition based on motor imagery tasks. PMID- 15876617 TI - Fractal dynamics of body motion in patients with Parkinson's disease. AB - In this paper, we assess the complexity (fractal measure) of body motion during walking in patients with Parkinson's disease. The body motion of 11 patients with Parkinson's disease and 10 healthy elderly subjects was recorded using a triaxial accelerometry technique. A triaxial accelerometer was attached to the lumbar region. An assessment of the complexity of body motion was made using a maximum likelihood-estimator-based fractal analysis method. Our data suggest that the fractal measures of the body motion of patients with Parkinson's disease are higher than those of healthy elderly subjects. These results were statistically different in the X (anteroposterior), Y (lateral) and Z (vertical) directions of body motion between patients with Parkinson's disease and the healthy elderly subjects (p < 0.01 in X and Z directions and p < 0.05 in Y direction). The complexity (fractal measure) of body motion can be useful to assess and monitor the output from the motor system during walking in clinical practice. PMID- 15876618 TI - Investigating the complexity of respiratory patterns during recovery from severe hypoxia. AB - Progressive hypoxemia in anesthetized, peripherally chemodenervated piglets results in initial depression of the phrenic neurogram (PN) culminating in phrenic silence and, eventually, gasping. These changes reverse after the 30 min reoxygenation (recovery) period. To determine if changes in the PN patterns correspond to changes in temporal patterning, we have used the approximate entropy (ApEn) method to examine the effects of maturation on the complexity of breathing patterns in chemodenervated, vagotomized and decerebrated piglets during severe hypoxia and reoxygenation. The phrenic neurogram in piglets was recorded during eupnea (normal breathing), severe hypoxia (gasping) and recovery from severe hypoxia in 31 piglets (2-35 days). Nonlinear dynamical analysis of the phrenic neurogram was performed using the ApEn method. The mean ApEn values for a recording of five consecutive breaths during eupnea, a few phrenic neurogram signals during gasping, the beginning of the recovery period, and five consecutive breaths at every 5 min interval for the 30 min recovery period were calculated. Our data suggest that gasping resulted in reduced duration of the phrenic neurogram, and the gasp-like patterns exist at the beginning of the recovery. But, the durations of phrenic neurograms during recovery were increased after 10 min postreoxygenation, but were restored 30 min post recovery. The ApEn (complexity) values of the phrenic neurogram during eupnea were higher than those of gasping and the early (the onset of) recovery from severe hypoxia (p < 0.01), but were not statistically different than 5 min post recovery regardless of the maturation stages. These results suggest that hypoxia results in a reversible reconfiguration of the central respiratory pattern generator. PMID- 15876619 TI - Single-trial lambda wave identification using a fuzzy inference system and predictive statistical diagnosis. AB - The aim of the study was to automate the identification of a saccade-related visual evoked potential (EP) called the lambda wave. The lambda waves were extracted from single trials of electroencephalogram (EEG) waveforms using independent component analysis (ICA). A trial was a set of EEG waveforms recorded from 64 scalp electrode locations while a saccade was performed. Forty saccade related EEG trials (recorded from four normal subjects) were used in the study. The number of waveforms per trial was reduced from 64 to 22 by pre-processing. The application of ICA to the resulting waveforms produced 880 components (i.e. 4 subjects x 10 trials per subject x 22 components per trial). The components were divided into 373 lambda and 507 nonlambda waves by visual inspection and then they were represented by one spatial and two temporal features. The classification performance of a Bayesian approach called predictive statistical diagnosis (PSD) was compared with that of a fuzzy logic approach called a fuzzy inference system (FIS). The outputs from the two classification approaches were then combined and the resulting discrimination accuracy was evaluated. For each approach, half the data from the lambda and nonlambda wave categories were used to determine the operating parameters of the classification schemes while the rest (i.e. the validation set) were used to evaluate their classification accuracies. The sensitivity and specificity values when the classification approaches were applied to the lambda wave validation data set were as follows: for the PSD 92.51% and 91.73% respectively, for the FIS 95.72% and 89.76% respectively, and for the combined FIS and PSD approach 97.33% and 97.24% respectively (classification threshold was 0.5). The devised signal processing techniques together with the classification approaches provided for an effective extraction and classification of the single-trial lambda waves. However, as only four subjects were included, it will be valuable to further evaluate the methods on a larger group of subjects. PMID- 15876620 TI - The spatial and temporal characteristics of the apparent-diffusion-coefficient dependent fMRI signal changes during visual stimulation. AB - The blood oxygenation level dependent (BOLD) contrast has been commonly used to detect fMRI signal. The majority of the BOLD signals are believed to arise from the venous and capillary networks. However, only those from the capillaries are spatially close to the neuronal activities, while the signals from large veins could be distant, rendering the overall localization inaccurate. In recent years, an alternative contrast using arterial spin labeled (ASL) perfusion imaging techniques has been proposed for predominant capillary sensitivity. Such acquisition methods, however, are intrinsically limited in temporal resolution and spatial coverage. Another contrast mechanism, free of such constraints, is based on the apparent diffusion coefficient (ADC) changes during brain activation using isotropic diffusion weighting. It has been shown that these changes are synchronized with brain activation and that they, as a whole, temporally precede BOLD activation, suggesting significant upstream arterial contribution. Moreover, the spatial overlaps between the upstream ADC and downstream BOLD activations are shown to be more localized in the capillaries, which are the temporal and spatial middle ground. In this paper, we sought to further investigate the temporal and spatial characteristics of ADC contrast with additional arterial signal suppression. Also, a pixel-based evaluation was performed in conjunction with the averaged global assessment. It was found that in addition to the known spatial discrepancy and global timing advance compared to the BOLD signal, the ADC activation endured significant temporal heterogeneities. Such fine spatial and temporal assessment could help characterize the exact signal sources of ADC contrast, and ultimately achieve exclusive capillary sensitivity. PMID- 15876621 TI - A versatile all-channel stimulator for electrode arrays, with real-time control. AB - Over the last few decades, technology to record through ever increasing numbers of electrodes has become available to electrophysiologists. For the study of distributed neural processing, however, the ability to stimulate through equal numbers of electrodes, and thus to attain bidirectional communication, is of paramount importance. Here, we present a stimulation system for multi-electrode arrays which interfaces with existing commercial recording hardware, and allows stimulation through any electrode in the array, with rapid switching between channels. The system is controlled through real-time Linux, making it extremely flexible: stimulation sequences can be constructed on-the-fly, and arbitrary stimulus waveforms can be used if desired. A key feature of this design is that it can be readily and inexpensively reproduced in other labs, since it interfaces to standard PC parallel ports and uses only off-the-shelf components. Moreover, adaptation for use with in vivo multi-electrode probes would be straightforward. In combination with our freely available data-acquisition software, MeaBench, this system can provide feedback stimulation in response to recorded action potentials within 15 ms. PMID- 15876622 TI - Control of phase synchronization of neuronal activity in the rat hippocampus. AB - Analysis of the synchronization mechanisms of neural activity is crucial to the understanding of the generation, propagation and control of epileptiform activity. Recently, phase synchronization (PS) analysis was applied to quantify the partial synchrony that exists in complex chaotic or noisy systems. In a previous study, we have shown that neural activity between two remotely located sites can be synchronized through a complete cut of the tissue by endogenous non synaptic signals. Therefore, it should be possible to apply signals to control PS. In this study, we test the hypothesis that stimulation amplitudes below excitation level (sub-threshold) can be used to control phase synchronization of two neural signals and we investigate the underlying mechanisms. PS of neuronal activity is first analysed in two coupled Rossler neuron models. Both synchronization and desynchronization could be generated with sub-threshold sinusoidal stimulation. Phase synchronization was then studied in in vitro brain slices. Neuronal activity between two sites was modulated by the application of small sinusoidal electric fields. PS between two remote sites could be achieved by the application of two identical waveforms while phase desynchronization of two close sites was generated by the application of a stimulus at a single site. These results show that sub-threshold stimuli are able to phase synchronize or desynchronize two networks and suggest that small signals could play an important role in normal neural activity and epilepsy. PMID- 15876623 TI - Elucidation of causal relationships for multi-sourced activities in the human brain by directed transinformation between time series of equivalent dipoles. AB - Visualizing the causal relationships among multi-sourced activities in the human brain non-invasively is important for the elucidation of the processing mechanism and for clinical application of the diagnosis of disease. We will show our preliminary results of estimating equivalent dipoles to show the multi-sourced brain activities and analyse directed transinformation through time series of dipoles with three-dimensional display. First, we estimated the equivalent dipoles from evoked potentials via a three-layered concentric spherical model and two-dipole estimation. Second, we analysed the directed transinformation between two time series of the first and second dipole moments by use of the derived two dipole positions and moments as the loci and magnitude of brain activities. Therefore, we obtained bi-directional information flow between the neuronal activities localized in three-dimensional space of the brain with respect to 21 ch somatosensory evoked potentials (SEPs). Our preliminary results can be interpreted as showing that the information, calculated based on our method, flows from the first dipole cluster located in the thalamus, to the second dipole cluster located in the somatosensory area. This does not go against the neurophysiological knowledge of SEPs that the activities move from the thalamus to the somatosensory area. With the above, we show the potential possibility of realizing the elucidation of causal relationships. PMID- 15876624 TI - A brain-computer interface using electrocorticographic signals in humans. AB - Brain-computer interfaces (BCIs) enable users to control devices with electroencephalographic (EEG) activity from the scalp or with single-neuron activity from within the brain. Both methods have disadvantages: EEG has limited resolution and requires extensive training, while single-neuron recording entails significant clinical risks and has limited stability. We demonstrate here for the first time that electrocorticographic (ECoG) activity recorded from the surface of the brain can enable users to control a one-dimensional computer cursor rapidly and accurately. We first identified ECoG signals that were associated with different types of motor and speech imagery. Over brief training periods of 3-24 min, four patients then used these signals to master closed-loop control and to achieve success rates of 74-100% in a one-dimensional binary task. In additional open-loop experiments, we found that ECoG signals at frequencies up to 180 Hz encoded substantial information about the direction of two-dimensional joystick movements. Our results suggest that an ECoG-based BCI could provide for people with severe motor disabilities a non-muscular communication and control option that is more powerful than EEG-based BCIs and is potentially more stable and less traumatic than BCIs that use electrodes penetrating the brain. PMID- 15876625 TI - Using human extra-cortical local field potentials to control a switch. AB - Individuals with profound paralysis and mutism require a communication channel. Traditional assistive technology devices eventually fail, especially in the case of amyotrophic lateral sclerosis (ALS) subjects who gradually become totally locked-in. A direct brain-to-computer interface that provides switch functions can provide a direct communication channel to the external world. Electroencephalographic (EEG) signals recorded from scalp electrodes are significantly degraded due to skull and scalp attenuation and ambient noise. The present system using conductive skull screws allows more reliable access to cortical local field potentials (LFPs) without entering the brain itself. We describe an almost locked-in human subject with ALS who activated a switch using online time domain detection techniques. Frequency domain analysis of his LFP activity demonstrates this to be an alternative method of detecting switch activation intentions. With this brain communicator system it is reasonable to expect that locked-in, but cognitively intact, humans will always be able to communicate. PMID- 15876626 TI - Topographically modified surfaces affect orientation and growth of hippocampal neurons. AB - Extracellular matrix molecules provide biochemical and topographical cues that influence cell growth in vivo and in vitro. Effects of topographical cues on hippocampal neuron growth were examined after 14 days in vitro. Neurons from hippocampi of rat embryos were grown on poly-L-lysine-coated silicon surfaces containing fields of pillars with varying geometries. Photolithography was used to fabricate 1 microm high pillar arrays with different widths and spacings. Beta(III)-tubulin and MAP-2 immunocytochemistry and scanning electron microscopy were used to describe neuronal processes. Automated two-dimensional tracing software quantified process orientation and length. Process growth on smooth surfaces was random, while growth on pillared surfaces exhibited the most faithful alignment to pillar geometries with smallest gap sizes. Neurite lengths were significantly longer on pillars with the smallest inter-pillar spacings (gaps) and 2 microm pillar widths. These data indicate that physical cues affect neuron growth, suggesting that extracellular matrix topography may contribute to cell growth and differentiation. These results demonstrate new strategies for directing and promoting neuronal growth that will facilitate studies of synapse formation and function and provide methods to establish defined neural networks. PMID- 15876627 TI - Enhanced peripheral nerve regeneration through a poled bioresorbable poly(lactic co-glycolic acid) guidance channel. AB - In this study we investigated the effects of materials prepared with electrical poling on neurite outgrowth in vitro and nerve regeneration in vivo. Neuro-2a cells were seeded on poled and unpoled poly(lactic-co-glycolic) (PLGA) films and observed at time periods 24, 48 and 72 h post-seeding. The percentage of cells with neurites and the neurites per cell were quantified using light microscopy. At 48 and 72 h post-seeding, both the number of cells with neurites and the neurites per cell were significantly increased on the poled films compared to those on unpoled films. An established rat sciatic nerve model was used for in vivo studies to assess the effects of PLGA guides, poled for two different periods, on peripheral nerve regeneration. Guides were inserted in rats to bridge a 1.0 cm gap created in the right sciatic nerve. After four weeks, nerves regenerated through poled guides displayed a significant increase in conduction velocity and significantly increased numbers of axons across the guides, as compared to nerves regenerating through an unpoled guidance channel. Electrical poling was shown to promote neurite growth, axon regeneration and the conduction rate of the repaired nerve. We concluded that guides prepared with electrical poling enhance peripheral nerve regeneration. PMID- 15876628 TI - Can standard surface EMG processing parameters be used to estimate motor unit global firing rate? AB - The relations between motor unit global firing rates and established quantitative measures for processing the surface electromyogram (EMG) signals were explored using a simulation approach. Surface EMG signals were simulated using the reported properties of the first dorsal interosseous muscle in man, and the models were varied systematically, using several hypothetical relations between motor unit electrical and force output, and also using different motor unit firing rate strategies. The utility of using different EMG processing parameters to help estimate global motor unit firing rate was evaluated based on their relations to the number of motor unit action potentials (MUAPs) in the simulated surface EMG signals. Our results indicate that the relation between motor unit electrical and mechanical properties, and the motor unit firing rate scheme are all important factors determining the form of the relation between surface EMG amplitude and motor unit global firing rate. Conversely, these factors have less impact on the relations between turn or zero-crossing point counts and the number of MUAPs in surface EMG. We observed that the number of turn or zero-crossing points tends to saturate with the increase in the MUAP number in surface EMG, limiting the utility of these measures as estimates of MUAP number. The simulation results also indicate that the mean or median frequency of the surface EMG power spectrum is a poor indicator of the global motor unit firing rate. PMID- 15876629 TI - Fractal dynamics of body motion in post-stroke hemiplegic patients during walking. AB - In this paper, we quantify the complexity of body motion during walking in post stroke hemiplegic patients. The body motion of patients and healthy elderly subjects was measured by using the accelerometry technique. The complexity of body motion was quantified using the maximum likelihood estimator (MLE-) based fractal analysis methods. Our results suggest that the fractal dimensions of the body motion in post-stroke hemiplegic patients at several Brunnstrom stages were significantly higher than those of healthy elderly subjects (p < 0.05). However, in the hemiplegic patients, the fractal dimensions were more related to Brunnstrom stages. PMID- 15876630 TI - Simulation of the effect of Na+ and Cl- on the velocity of a spreading depression wave using a simplified electrochemical model of synaptic terminals. AB - In the study of the spreading depression (SD) wave phenomenon and its dynamics, it is necessary to describe the ionic movements along the extracellular space, as well as between this and the intracellular space. In both cases, the ionic movement includes a double coupling involving the concentration and the potential gradients and hence must be described by electrodiffusion mechanisms. Based on this, the effects of the ionic composition on the characteristics of the wave propagation can be predicted. The influence of varying extracellular sodium and chloride concentrations on the velocity of propagation of the SD wave was investigated by simulation. The results achieved are close to the experimental measurement from the literature. These findings suggest the potentiality of the model proposed in supporting the interpretation of experimental data in neuronal tissues, particularly the SD. PMID- 15876631 TI - Estimation of neural energy in microelectrode signals. AB - We considered the problem of determining the neural contribution to the signal recorded by an intracortical electrode. We developed a linear least-squares approach to determine the energy fraction of a signal attributable to an arbitrary number of autocorrelation-defined signals buried in noise. Application of the method requires estimation of autocorrelation functions R(ap)(tau) characterizing the action potential (AP) waveforms and R(n)(tau) characterizing background noise. This method was applied to the analysis of chronically implanted microelectrode signals from motor cortex of rat. We found that neural (AP) energy consisted of a large-signal component which grows linearly with the number of threshold-detected neural events and a small-signal component unrelated to the count of threshold-detected AP signals. The addition of pseudorandom noise to electrode signals demonstrated the algorithm's effectiveness for a wide range of noise-to-signal energy ratios (0.08 to 39). We suggest, therefore, that the method could be of use in providing a measure of neural response in situations where clearly identified spike waveforms cannot be isolated, or in providing an additional 'background' measure of microelectrode neural activity to supplement the traditional AP spike count. PMID- 15876632 TI - Motor imagery classification by means of source analysis for brain-computer interface applications. AB - We report a pilot study of performing classification of motor imagery for brain computer interface applications, by means of source analysis of scalp-recorded EEGs. Independent component analysis (ICA) was used as a spatio-temporal filter extracting signal components relevant to left or right motor imagery (MI) tasks. Source analysis methods including equivalent dipole analysis and cortical current density imaging were applied to reconstruct equivalent neural sources corresponding to MI, and classification was performed based on the inverse solutions. The classification was considered correct if the equivalent source was found over the motor cortex in the corresponding hemisphere. A classification rate of about 80% was achieved in the human subject studied using both the equivalent dipole analysis and the cortical current density imaging analysis. The present promising results suggest that the source analysis approach could manifest a clearer picture on the cortical activity, and thus facilitate the classification of MI tasks from scalp EEGs. PMID- 15876633 TI - Neural network classification of autoregressive features from electroencephalogram signals for brain-computer interface design. AB - In this paper, we have designed a two-state brain-computer interface (BCI) using neural network (NN) classification of autoregressive (AR) features from electroencephalogram (EEG) signals extracted during mental tasks. The main purpose of the study is to use Keirn and Aunon's data to investigate the performance of different mental task combinations and different AR features for BCI design for individual subjects. In the experimental study, EEG signals from five mental tasks were recorded from four subjects. Different combinations of two mental tasks were studied for each subject. Six different feature extraction methods were used to extract the features from the EEG signals: AR coefficients computed with Burg's algorithm, AR coefficients computed with a least-squares (LS) algorithm and adaptive autoregressive (AAR) coefficients computed with a least-mean-square (LMS) algorithm. All the methods used order six applied to 125 data points and these three methods were repeated with the same data but with segmentation into five segments in increments of 25 data points. The multilayer perceptron NN trained by the back-propagation algorithm (MLP-BP) and linear discriminant analysis (LDA) were used to classify the computed features into different categories that represent the mental tasks. We compared the classification performances among the six different feature extraction methods. The results showed that sixth-order AR coefficients with the LS algorithm without segmentation gave the best performance (93.10%) using MLP-BP and (97.00%) using LDA. The results also showed that the segmentation and AAR methods are not suitable for this set of EEG signals. We conclude that, for different subjects, the best mental task combinations are different and proper selection of mental tasks and feature extraction methods are essential for the BCI design. PMID- 15876634 TI - Synergistic effects of micropatterned biodegradable conduits and Schwann cells on sciatic nerve regeneration. AB - This paper describes a novel biodegradable conduit that provides a combination of physical, chemical and biological cues at the cellular level to facilitate peripheral nerve regeneration. The conduit consists of a porous poly(D,L-lactic acid) (PDLLA) tubular support structure with a micropatterned inner lumen. Schwann cells were pre-seeded into the lumen to provide additional trophic support. Conduits with micropatterned inner lumens pre-seeded with Schwann cells (MS) were fabricated and compared with three types of conduits used as controls: M (conduits with micropatterned inner lumens without pre-seeded Schwann cells), NS (conduits without micropatterned inner lumens pre-seeded with Schwann cells) and N (conduits without micropatterned inner lumens, without pre-seeded Schwann cells). The conduits were implanted in rats with 1 cm sciatic nerve transections and the regeneration and functional recovery were compared in the four different cases. The number or size of regenerated axons did not vary significantly among the different conduits. The time of recovery, and the sciatic function index, however, were significantly enhanced using the MS conduits, based on qualitative observations as well as quantitative measurements using walking track analysis. This demonstrates that biodegradable micropatterned conduits pre-seeded with Schwann cells that provide a combination of physical, chemical and biological guidance cues for regenerating axons at the cellular level offer a better alternative for repairing sciatic nerve transactions than conventional biodegradable conduits. PMID- 15876635 TI - A synaptic input portal for a mapped clock oscillator model of neuronal electrical rhythmic activity. AB - Neuronal electrical oscillations play a central role in a variety of situations, such as epilepsy and learning. The mapped clock oscillator (MCO) model is a general model of transmembrane voltage oscillations in excitable cells. In order to be able to investigate the behaviour of neuronal oscillator populations, we present a neuronal version of the model. The neuronal MCO includes an extra input portal, the synaptic portal, which can reflect the biological relationships in a chemical synapse between the frequency of the presynaptic action potentials and the postsynaptic resting level, which in turn affects the frequency of the postsynaptic potentials. We propose that the synaptic input-output relationship must include a power function in order to be able to reproduce physiological behaviour such as resting level saturation. One linear and two power functions (Butterworth and sigmoidal) are investigated, using the case of an inhibitory synapse. The linear relation was not able to produce physiologically plausible behaviour, whereas both the power function examples were appropriate. The resulting neuronal MCO model can be tailored to a variety of neuronal cell types, and can be used to investigate complex population behaviour, such as the influence of network topology and stochastic resonance. PMID- 15876636 TI - Adaptive autoregressive identification with spectral power decomposition for studying movement-related activity in scalp EEG signals and basal ganglia local field potentials. AB - We propose a method that combines adaptive autoregressive (AAR) identification and spectral power decomposition for the study of movement-related spectral changes in scalp EEG signals and basal ganglia local field potentials (LFPs). This approach introduces the concept of movement-related poles, allowing one to study not only the classical event-related desynchronizations (ERD) and synchronizations (ERS), which correspond to modulations of power, but also event related modulations of frequency. We applied the method to analyze movement related EEG signals and LFPs contemporarily recorded from the sensorimotor cortex, the globus pallidus internus (GPi) and the subthalamic nucleus (STN) in a patient with Parkinson's disease who underwent stereotactic neurosurgery for the implant of deep brain stimulation (DBS) electrodes. In the AAR identification we compared the whale and the exponential forgetting factors, showing that the whale forgetting provides a better disturbance rejection and it is therefore more suitable to investigate movement-related brain activity. Movement-related power modulations were consistent with previous studies. In addition, movement-related frequency modulations were observed from both scalp EEG signals and basal ganglia LFPs. The method therefore represents an effective approach to the study of movement-related brain activity. PMID- 15876637 TI - Oscillations in the power spectra of motor unit signals caused by refractoriness variations. AB - The refractory period of a motor unit is an important mechanism that regulates the motor unit firing, and its variation has been found in many physiological cases. In this study, a new observation that an increase in the motor unit refractoriness results in an enhancement of oscillations, or ripple effects, in the motor unit output power density spectra (PDS) has been identified and studied. The effects of the refractoriness variation on the PDS of motor unit firing were investigated on three levels: theoretical modeling, simulation and electromyographic (EMG) experimentation on human subjects. Both theoretical modeling and simulation showed the enhanced oscillations, ripple effects, in MUAPT PDS, given the increase in the refractoriness. It was also found that the extent of the increment in output PDS oscillation could be related to the motor unit size and the mean firing rate of the stimulation. A needle EMG experiment on biceps brachii muscles of five healthy human subjects was carried out during isometric contraction at 20% maximum voluntary contraction (MVC) for 20 s with a fatigue effort proceeded by MVC. The increased oscillations in the PDS of the real MUAPTs were observed with the rising of the motor unit refractoriness due to fatigue. The study gives new information for EMG spectra interpretation, and also provides a potential method for accessing neuromuscular transmission failure (NTF) due to fatigue during voluntary contraction. PMID- 15876638 TI - Analysis of real-time numerical integration methods applied to dynamic clamp experiments. AB - Real-time systems are frequently used as an experimental tool, whereby simulated models interact in real time with neurophysiological experiments. The most demanding of these techniques is known as the dynamic clamp, where simulated ion channel conductances are artificially injected into a neuron via intracellular electrodes for measurement and stimulation. Methodologies for implementing the numerical integration of the gating variables in real time typically employ first order numerical methods, either Euler or exponential Euler (EE). EE is often used for rapidly integrating ion channel gating variables. We find via simulation studies that for small time steps, both methods are comparable, but at larger time steps, EE performs worse than Euler. We derive error bounds for both methods, and find that the error can be characterized in terms of two ratios: time step over time constant, and voltage measurement error over the slope factor of the steady-state activation curve of the voltage-dependent gating variable. These ratios reliably bound the simulation error and yield results consistent with the simulation analysis. Our bounds quantitatively illustrate how measurement error restricts the accuracy that can be obtained by using smaller step sizes. Finally, we demonstrate that Euler can be computed with identical computational efficiency as EE. PMID- 15876639 TI - Estimation of temporary change of activation areas by moving an analysis time window in fMRI measurement. AB - In this paper, we propose a method to acquire temporal changes of activations by moving an analysis time window. An advantage of this method is that it can acquire rough changes of activated areas even with the data having low time resolution. We ascertained that activations from our method do not contradict previous reports on the oddball paradigm, thus showing its effectiveness. Eight normal subjects participated in the study, which consisted of a random series of 30 target and 70 nontarget stimuli. We investigated the activated area in three kinds of analysis time sections, from stimulus onset to 5 s after the stimulus (time section A), from 2 to 7 s after (B) and from 4 to 9 s after (C). In time section A, representative activated areas were regions including the left and supplementary motor areas (SMA), and cerebellum. In B, regions including the left motor area and SMA, right parahippocampal gyrus (Broadmann Area (BA) 30), right limbic lobe and cerebellum were activated. In C, bilaterally postcentral gyrus (BA 3,40), right anterior cingulate (ACC, BA 32), left middle frontal gyrus (BA 9) and right parahippocampal gyrus were activated. Most activations were consistent with previous studies. PMID- 15876640 TI - Extracellular voltage profile for reversing the recruitment order of peripheral nerve stimulation: a simulation study. AB - Electrical stimulation of peripheral nerve activates large-diameter fibers before small ones. A physiological recruitment order, from small to large-diameter axons, is desirable in many applications. Previous studies using computer simulations showed that selective activation of small fibers could be achieved by reshaping the extracellular voltage profile along the nerve using an array of nine electrodes. In this study, several electrode-array configurations were tested in order to minimize the number of contacts. Electrode arrays of 5, 7, 9, and 11 contacts with 0.75 mm contact separation were performed in computer simulations of dog sacral root (S2). Electrode arrays of 5 and 7 contacts recruited 40% of small axons (<10 microm) when recruiting only 10% of larger axons. Effectiveness of 9- and 11-contact arrays decreased with the presence of epineurium and perineurium. The effectiveness of electrode arrays was independent of stimulation pulsewidth. The biphasic-pulse stimulation with the amplitude of the second phase set as low as possible should be used to prevent the excitation of large axons during the second phase and to minimize the electrode corrosion. Arrays of 5 and 7 contacts also decreased the recruitment curve slope to 26% and 51% of the tripolar electrode, respectively. This modeling study predicts that reversing the recruitment order of peripheral nerve stimulation could be achieved by reshaping the extracellular voltage using electrode arrays of 5 or 7 contacts. PMID- 15876641 TI - A new approach in the BCI research based on fractal dimension as feature and Adaboost as classifier. AB - High rate classification of imagery tasks is still one of the hot topics among the brain computer interface (BCI) groups. In order to improve this rate, a new approach based on fractal dimension as feature and Adaboost as classifier is presented for five subjects in this paper. To have a comparison, features such as band power, Hjorth parameters along with LDA classifier have been taken into account. Fractal dimension as a feature with Adaboost and LDA can be considered as alternative combinations for BCI applications. PMID- 15876642 TI - An in vitro model for investigating impedance changes with cell growth and electrical stimulation: implications for cochlear implants. AB - The impedance of stimulating electrodes used in cochlear implants and other neural prostheses often increases post-implantation, and is thought to be due to fibrous tissue encapsulation of the electrode array. Increased impedance results in higher power requirements to stimulate target neurons at set charge densities. We developed an in vitro model to investigate the electrode-tissue interface in a highly controlled environment. This model was tested using three cell types, with and without charge-balanced biphasic electrical stimulation. Under standard tissue culture conditions, a monolayer of cells was grown over the electrode surface. Electrode impedance increased in proportion to the extent of cell coverage of the electrode. Cell type was a significant factor in the amount of impedance increase, with kidney epithelial cells (MDCK) creating the greatest impedance, followed by dissociated rat skin fibroblasts and then macrophages (J774). The application of electrical stimulation to cell-covered electrodes caused impedance fluctuations similar to that seen in vivo, with a lowering of impedance immediately following stimulation, and a recovery to pre-stimulation levels during inactive periods. Examination of these electrodes suggests that the stimulation-induced impedance changes were due to the amount of cell cover over the electrodes. This in vitro technique accurately models the changes in impedance observed with neural prostheses in vivo, and shows the close relationship between impedance and tissue coverage adjacent to the electrode surface. We believe that this in vitro approach holds great promise to further our knowledge of the mechanisms contributing to electrode impedance. PMID- 15876643 TI - Determining the receptive field of a neural filter. AB - In this paper, a method for determining the receptive field and the structure of hidden layers of a neural filter (NF) was developed and evaluated. With the proposed method, redundant units are removed from input and hidden layers in an NF based on the influence of removal of units on the error between output and teaching images. By performing the removal of units and retraining for recovery of the loss of the removal repeatedly, the receptive field and a reduced structure of hidden layers are determined. Experiments with NFs were performed for acquiring the function of a known filter, for the reduction of noise in natural images and for the reduction of noise in medical image sequences. By use of the proposed method, redundant units were able to be removed from NFs, while the performance of the NFs was maintained. Experimental results suggested that, with the proposed method, a reasonable receptive field for a given image processing task could be determined, i.e., the receptive field of the NF trained to obtain the function of a filter corresponded to the kernel of the filter, and the receptive fields of the NFs for noise reduction gathered around the object pixels in the input regions of the NFs. PMID- 15876644 TI - An evaluation of the utility and limitations of counting motor unit action potentials in the surface electromyogram. AB - The number of motor unit action potentials (MUAPs) appearing in the surface electromyogram (EMG) signal is directly related to motor unit recruitment and firing rates and therefore offers potentially valuable information about the level of activation of the motoneuron pool. In this paper, based on morphological features of the surface MUAPs, we try to estimate the number of MUAPs present in the surface EMG by counting the negative peaks in the signal. Several signal processing procedures are applied to the surface EMG to facilitate this peak counting process. The MUAP number estimation performance by this approach is first illustrated using the surface EMG simulations. Then, by evaluating the peak counting results from the EMG records detected by a very selective surface electrode, at different contraction levels of the first dorsal interosseous (FDI) muscles, the utility and limitations of such direct peak counts for MUAP number estimation in surface EMG are further explored. PMID- 15876645 TI - The FDA's role in medical device clinical studies of human subjects. AB - This paper provides an overview of the United States Food and Drug Administration's (FDA) role as a regulatory agency in medical device clinical studies involving human subjects. The FDA's regulations and responsibilities are explained and the device application process discussed. The specific medical device regulatory authorities are described as they apply to the development and clinical study of retinal visual prosthetic devices. The FDA medical device regulations regarding clinical studies of human subjects are intended to safeguard the rights and safety of subjects. The data gathered in pre-approval clinical studies provide a basis of valid scientific evidence in order to demonstrate the safety and effectiveness of a medical device. The importance of a working understanding of applicable medical device regulations from the beginning of the device development project is emphasized particularly for novel, complex products such as implantable visual prosthetic devices. PMID- 15876646 TI - Design of a high-resolution optoelectronic retinal prosthesis. AB - It has been demonstrated that electrical stimulation of the retina can produce visual percepts in blind patients suffering from macular degeneration and retinitis pigmentosa. However, current retinal implants provide very low resolution (just a few electrodes), whereas at least several thousand pixels would be required for functional restoration of sight. This paper presents the design of an optoelectronic retinal prosthetic system with a stimulating pixel density of up to 2500 pix mm(-2) (corresponding geometrically to a maximum visual acuity of 20/80). Requirements on proximity of neural cells to the stimulation electrodes are described as a function of the desired resolution. Two basic geometries of sub-retinal implants providing required proximity are presented: perforated membranes and protruding electrode arrays. To provide for natural eye scanning of the scene, rather than scanning with a head-mounted camera, the system operates similar to 'virtual reality' devices. An image from a video camera is projected by a goggle-mounted collimated infrared LED-LCD display onto the retina, activating an array of powered photodiodes in the retinal implant. The goggles are transparent to visible light, thus allowing for the simultaneous use of remaining natural vision along with prosthetic stimulation. Optical delivery of visual information to the implant allows for real-time image processing adjustable to retinal architecture, as well as flexible control of image processing algorithms and stimulation parameters. PMID- 15876647 TI - Fabrication of implantable microelectrode arrays by laser cutting of silicone rubber and platinum foil. AB - A new method for fabrication of microelectrode arrays comprised of traditional implant materials is presented. The main construction principle is the use of spun-on medical grade silicone rubber as insulating substrate material and platinum foil as conductor (tracks, pads and electrodes). The silicone rubber and the platinum foil are patterned by laser cutting using an Nd:YAG laser and a microcontroller-driven, stepper-motor operated x-y table. The method does not require expensive clean room facilities and offers an extremely short design-to prototype time of below 1 day. First prototypes demonstrate a minimal achievable feature size of about 30 microm. PMID- 15876648 TI - A method and technical equipment for an acute human trial to evaluate retinal implant technology. AB - This paper reports on methods and technical equipment to investigate the epiretinal stimulation of the retina in blind human subjects in acute trials. Current is applied to the retina through a thin, flexible microcontact film (microelectrode array) with electrode diameters ranging from 50 to 360 microm. The film is mounted in a custom-designed surgical tool that is hand-held by the surgeon during stimulation. The eventual goal of the work is the development of a chronically implantable retinal prosthesis to restore a useful level of vision to patients who are blind with outer retinal degenerations, specifically retinitis pigmentosa and macular degeneration. PMID- 15876649 TI - Visual acuity measurement of prosthetic vision: a virtual-reality simulation study. AB - A virtual-reality simulation tested prosthetic visual acuity for both rectangular and hexagonal phosphene grids. Thirteen normally sighted, untrained subjects were required to identify the Landolt C optotype over ten sessions. Overall performance, performance by filter setting (image analysis), and performance by size and orientation of the Landolt C optotype are reported. The results indicated that the hexagonal grid had a slight (4.1%) but statistically significant (p < 0.0001) performance advantage over the rectangular grid for correct identification of the testing symbol. It was also observed that both the phosphene grids had distinct performance profiles relating to their respective spatial orientation. Over the course of the ten sessions, learning afforded subjects an averaged improved performance of 10%. PMID- 15876650 TI - Responses of rabbit retinal ganglion cells to electrical stimulation with an epiretinal electrode. AB - Rational selection of electrical stimulus parameters for an electronic retinal prosthesis requires knowledge of the electrophysiological responses of retinal neurons to electrical stimuli. In this study, we examined the effects of cathodal and anodal current pulses on the extracellularly recorded responses of OFF and ON rabbit retinal ganglion cells (RGCs) in an in vitro preparation. Current pulses (1 msec duration), delivered by a 125 microm electrode placed on the inner retinal surface within the receptive field of a RGC, produced both short-latency (< or =5 msec) and long-latency (8-60 msec) responses. The long-latency responses, but not the short-latency responses, were abolished upon application of the glutamate receptor antagonists CNQX and NBQX, thus indicating that the long-latency responses of RGCs are due to activation of presynaptic neurons in the retina. The latency of the long-latency response depended upon the polarity of the stimulus. For OFF RGCs, the average latency was 11 msec for a cathodal stimulus and 24 msec for an anodal stimulus. For ON RGCs, the average latency was 25 msec for a cathodal stimulus and 16 msec for an anodal stimulus. The threshold current also depended upon the polarity of the stimulus, at least for OFF RGCs. The average threshold current for evoking a long-latency response in OFF RGCs was 10 microA for a cathodal stimulus and 21 microA for an anodal stimulus. In ON RGCs, the average threshold current was 13 microA for a cathodal stimulus and 15 microA for an anodal stimulus. PMID- 15876651 TI - Creating a meaningful visual perception in blind volunteers by optic nerve stimulation. AB - A blind volunteer, suffering from retinitis pigmentosa, has been chronically implanted with an optic nerve visual prosthesis. Vision rehabilitation with this volunteer has concentrated on the development of a stimulation strategy according to which video camera images are converted into stimulation pulses. The aim is to convey as much information as possible about the visual scene within the limits of the device's capabilities. Pattern recognition tasks were used to assess the effectiveness of the stimulation strategy. The results demonstrate how even a relatively basic algorithm can efficiently convey useful information regarding the visual scene. By increasing the number of phosphenes used in the algorithm, better performance is observed but a longer training period is required. After a learning period, the volunteer achieved a pattern recognition score of 85% at 54 s on average per pattern. After nine evaluation sessions, when using a stimulation strategy exploiting all available phosphenes, no saturation effect has yet been observed. PMID- 15876652 TI - A preparation for studying electrical stimulation of the retina in vivo in rat. AB - A remaining challenge to the development of electronic prostheses for vision is improving the effectiveness of retinal stimulation. Electrode design and stimulus parameters need to be optimized such that the neural output from the retina conveys information to the mind's eye that aids the patient in interpreting his or her environment. This optimization will require a detailed understanding of the response of the retina to electrical stimulation. The identity and response characteristics of the cellular targets of stimulation need to be defined and evaluated. Described here is an in vivo preparation for studying electrical stimulation of the retina in rat at the cellular level. The use of rat makes available a number of well-described models of retinal disease that motivate prosthesis development. Artificial stimulation can be investigated by adapting techniques traditionally employed to study the response of the retina to photic stimuli, such as recording at the cornea, single-cell recording, and pharmacological dissection of the response. Pilot studies include amplitude intensity response data for subretinal and transretinal stimulation paradigms recorded in wild-type rats and a transgenic rat model of autosomal dominant retinitis pigmentosa. The ability to record single-unit ganglion cell activity in vivo is also demonstrated. PMID- 15876653 TI - Possible sources of neuroprotection following subretinal silicon chip implantation in RCS rats. AB - Current retinal prosthetics are designed to stimulate existing neural circuits in diseased retinas to create a visual signal. However, implantation of retinal prosthetics may create a neurotrophic environment that also leads to improvements in visual function. Possible sources of increased neuroprotective effects on the retina may arise from electrical activity generated by the prosthetic, mechanical injury due to surgical implantation, and/or presence of a chronic foreign body. This study evaluates these three neuroprotective sources by implanting Royal College of Surgeons (RCS) rats, a model of retinitis pigmentosa, with a subretinal implant at an early stage of photoreceptor degeneration. Treatment groups included rats implanted with active and inactive devices, as well as sham operated. These groups were compared to unoperated controls. Evaluation of retinal function throughout an 18 week post-implantation period demonstrated transient functional improvements in eyes implanted with an inactive device at 6, 12 and 14 weeks post-implantation. However, the number of photoreceptors located directly over or around the implant or sham incision was significantly increased in eyes implanted with an active or inactive device or sham-operated. These results indicate that in the RCS rat localized neuroprotection of photoreceptors from mechanical injury or a chronic foreign body may provide similar results to subretinal electrical stimulation at the current output evaluated here. PMID- 15876654 TI - Comparison of electrically evoked cortical potential thresholds generated with subretinal or suprachoroidal placement of a microelectrode array in the rabbit. AB - The aim of the study was to directly compare the threshold electrical charge density of the retina (retinal threshold) in rabbits for the generation of electrical evoked potentials (EEP) by delivering electrical stimulation with a custom-made microelectrode array (MEA) implanted into either the subretinal or suprachoroidal space. Nine eyes of seven Dutch-belted rabbits were studied. The electroretinogram (ERG), visual evoked potentials (VEP) and EEP were recorded. Electrodes for the VEP and EEP were placed on the dura mater overlying the visual cortex. The EEP was recorded following electrical stimulation of the MEA placed either subretinally beneath the visual streak of the retina or in the suprachoroidal space in the rabbit eye. An ab externo approach was used for placement of the MEA. Liquid perfluorodecaline (PFCL; 0.4 ml) was placed within the vitreous cavity to flatten the neurosensory retina on the MEA after subretinal implantation. The retinal threshold for generation of an EEP was determined for each MEA placement by three consecutive measurements consisting of 100 computer-averaged recordings. Animals were sacrificed at the conclusion of the experiment and the eyes were enucleated for histological examination. The retinal threshold to generate an EEP was 9 +/- 7 nC (0.023 +/- 0.016 mC cm(-2)) within the subretinal space and 150 +/- 122 nC (0.375 +/- 0.306 mC cm(-2)) within the suprachoroidal space. Histology showed disruption of the outer retina with subretinal but not suprachoroidal placement. The retinal threshold to elicit an EEP is significantly lower with subretinal placement of the MEA compared to suprachoroidal placement (P < 0.05). The retinal threshold charge density with a subretinal MEA is well below the published charge limit of 1 mC cm(-2), which is the level below which chronic stimulation of the retina is considered necessary to avoid tissue damage (Shannon 1992 IEEE Trans. Biomed. Eng. 39 424-6). PMID- 15876655 TI - In vitro activation of retinal cells: estimating location of stimulated cell by using a mathematical model. AB - Activation of neurons at different depths within the retina and at various eccentricities from the stimulating electrode will presumably influence the visual percepts created by a retinal prosthesis. With an electrical prosthesis, neurons will be activated in relation to the stimulating charge that impacts their cell membranes. The common model used to predict charge density is Coulomb's law, also known as the square law. We propose a modified model that can be used to predict neuronal depth that takes into account: (1) finite dimensions related to the position and size of the stimulating and return electrodes and (2) two-dimensional displacements of neurons with respect to the electrodes, two factors that are not considered in the square law model. We tested our model by using in vitro physiological threshold data that we had obtained previously for eight OFF-center brisk-transient rabbit retinal ganglion cells. For our most spatially dense threshold data (25 microm increments up to 100 microm from the cell body), our model estimated the depth of one RGC to be 76 +/- 76 microm versus 87 +/- 62 microm (median: SD) for the square law model, respectively. This difference was not statistically significant. For the seven other RGCs for which we had obtained threshold data up to 800 microm from the cell body, the estimate of the RGC depth (using data obtained along the X axis) was 96 +/- 74 versus 20 +/- 20 microm for the square law and our modified model, respectively. Although this difference was not statistically significant (Student t-test: p = 0.12), our model provided median values much closer to the estimated depth of these RGCs (>>25 microm). This more realistic estimate of cell depth predicted by our model is not unexpected in this latter data set because of the more spatially distributed threshold data points that were evaluated. Our model has theoretical advantages over the traditional square law model under certain conditions, especially when considering neurons that are horizontally displaced from the stimulating electrode. Our model would have to be tested with a larger threshold data pool to permit more conclusive statements about the relative value of our model versus the traditional square law model under special circumstances. PMID- 15876656 TI - Transscleral implantation and neurophysiological testing of subretinal polyimide film electrodes in the domestic pig in visual prosthesis development. AB - Loss of photoreceptor function is responsible for a variety of blinding diseases, including retinitis pigmentosa. Advances in microtechnology have led to the development of electronic visual prostheses which are currently under investigation for the treatment of human blindness. The design of a subretinal prosthesis requires that the stimulation device should be implantable in the subretinal space of the eye. Current limitations in eye surgery have to be overcome to demonstrate the feasibility of this approach and to determine basic stimulation parameters. Therefore, polyimide film-bound electrodes were implanted in the subretinal space in anaesthetized domestic pigs as a prelude to electrical stimulation in acute experiments. Eight eyes underwent surgery to demonstrate the transscleral implantability of the device. Four of the eight eyes were stimulated electrically. In these four animals the cranium was prepared for epidural recording of evoked visual cortex responses, and stimulation was performed with sequences of current impulses. All eight subretinal implantation procedures were carried out successfully with polyimide film electrodes and each electrode was implanted beneath the outer retina of the posterior pole of the operated eyes. Four eyes were used for neurophysiological testing, involving recordings of epidural cortical responses to light and electrical stimulation. A light stimulus response, which occurred 40 ms after stimulation, proved the integrity of the operated eye. The electrical stimuli occurred about 20 ms after the onset of stimulation. The stimulation threshold was approximately 100 microA. Both the threshold and the cortical responses depended on the correspondence between retinal stimulation and cortical recording sites and on the number of stimulation electrodes used simultaneously. The subretinal implantation of complex stimulation devices using the transscleral procedure with consecutive subretinal stimulation is feasible in acute experiments in an animal model approximating to the situation in humans. The domestic pig is an appropriate animal model for basic testing of subretinal implants. Animal experiments with chronically implanted devices and long-term stimulation are advisable to prepare the field for successful human experiments. PMID- 15876657 TI - Long-term stimulation by active epiretinal implants in normal and RCD1 dogs. AB - An epiretinal prosthesis, consisting of an extraocular microelectronic stimulator and an intraocular electrode array, was implanted in one eye of three blind and three sighted dogs. Three dogs (2 blind, 1 normal) were stimulated for 120 days, and two dogs (both normal) for 60 and 103 days respectively for 8-10 h/day at levels of 0.1 mC cm(-2) and 0.05 mC cm(-2), with each stimulus level presented to half of the array. One blind dog was kept as an inactive implant control. During the study period, electroretinograms (ERG) and fundus photographs were recorded. At the end of the study period, the dogs were sacrificed and histological and morphometric evaluation was made of the retina. No inflammatory reaction, neovascularization or hemorrhage was observed during the follow-up examinations. ERGs were unchanged. Stimulus levels used were of sufficient amplitude to elicit cortical evoked potentials. Histological evaluation showed no inflammatory infiltrates or changes in retina morphometry related to electrical stimulation when compared to the unstimulated control eye. Morphometric analysis revealed no consistent differences relating to electrical stimulation. In summary, chronic electrical stimulation of the dog retina at up to 0.1 mC cm(-2) with an epiretinal prosthesis does not appear to adversely affect the retina. PMID- 15876658 TI - How the retinal network reacts to epiretinal stimulation to form the prosthetic visual input to the cortex. AB - We considered the problem of determining how the retinal network may interact with electrical epiretinal stimulation in shaping the spike trains of ON and OFF ganglion cells, and thus the synaptic input to first-stage cortical neurons. To do so, we developed a biophysical model of the retinal network with nine stacked neuronal mosaics. Here, we describe the model's behavior under (i) electrical stimulation of a retina with complete cone photoreceptor loss, but an otherwise intact circuitry and (ii) electrical stimulation of a fully-functional retina. Our results show that electrical stimulation alone results in indiscriminate excitation of ON and OFF ganglion cells and a patchy input to the cortex with islands of excitation among regions of no net excitation. Activation of the retinal network biases the excitation of ON relative to OFF ganglion cells, and in addition, gradually interpolates and focuses the initial, patchy synaptic input to the cortex. As stimulation level increases, the cortical input spreads beyond the area occupied by the electrode contact. Further, at very strong stimulation levels, ganglion cell responses begin to saturate, resulting in a significant distortion in the spatial profile of the cortical input. These findings occur in both the normal and the degenerated retina simulations, but the normal retina exhibits a tighter spatiotemporal response. The complex spatiotemporal dynamics of the prosthetic input to the cortex that are revealed by our model should be addressed by prosthetic image encoders and by studies that simulate prosthetic vision. PMID- 15876659 TI - Tunable retina encoders for retina implants: why and how. AB - Current research towards retina implants for partial restoration of vision in blind humans with retinal degenerative dysfunctions focuses on implant and stimulation experiments and technologies. In contrast, our approach takes the availability of an epiretinal multi-electrode neural interface for granted and studies the conditions for successful joint information processing of both retinal prosthesis and brain. Our proposed learning retina encoder (RE) includes information processing modules to simulate the complex mapping operation of parts of the 5-layered neural retina and to provide an iterative, perception-based dialog between RE and human subject. Alternative information processing technologies in the learning RE are being described, which allow an individual optimization of the RE mapping operation by means of iterative tuning with learning algorithms in a dialog between implant wearing subject and RE. The primate visual system is modeled by a retina module (RM) composed of spatio temporal (ST) filters and a central visual system module (VM). RM performs a mapping 1 of an optical pattern P1 in the physical domain onto a retinal output vector R1(t) in a neural domain, whereas VM performs a mapping 2 of R1(t) in a neural domain onto a visual percept P2 in the perceptual domain. Retinal ganglion cell properties represent non-invertible ST filters in RE, which generate ambiguous output signals. VM generates visual percepts only if the corresponding R1(t) is properly encoded, contains sufficient information, and can be disambiguated. Based on the learning RE and the proposed visual system model, a novel retina encoder (RE*) is proposed, which considers both ambiguity removal and miniature eye movements during fixation. Our simulation results suggest that VM requires miniature eye movements under control of the visual system to retrieve unambiguous patterns P2 corresponding to P1. For retina implant applications, RE* can be tuned to generate optimal ganglion cell codes for epiretinal stimulation. PMID- 15876660 TI - Assessment of rigid multi-modality image registration consistency using the multiple sub-volume registration (MSR) method. AB - Registration of different imaging modalities such as CT, MRI, functional MRI (fMRI), positron (PET) and single photon (SPECT) emission tomography is used in many clinical applications. Determining the quality of any automatic registration procedure has been a challenging part because no gold standard is available to evaluate the registration. In this note we present a method, called the 'multiple sub-volume registration' (MSR) method, for assessing the consistency of a rigid registration. This is done by registering sub-images of one data set on the other data set, performing a crude non-rigid registration. By analysing the deviations (local deformations) of the sub-volume registrations from the full registration we get a measure of the consistency of the rigid registration. Registration of 15 data sets which include CT, MR and PET images for brain, head and neck, cervix, prostate and lung was performed utilizing a rigid body registration with normalized mutual information as the similarity measure. The resulting registrations were classified as good or bad by visual inspection. The resulting registrations were also classified using our MSR method. The results of our MSR method agree with the classification obtained from visual inspection for all cases (p < 0.02 based on ANOVA of the good and bad groups). The proposed method is independent of the registration algorithm and similarity measure. It can be used for multi-modality image data sets and different anatomic sites of the patient. PMID- 15876661 TI - An inter-laboratory comparison study of image quality of PET scanners using the NEMA NU 2-2001 procedure for assessment of image quality. AB - An inter-laboratory comparison study was conducted to assess the image quality of PET scanners in Austria. The survey included both dedicated PET scanners (D-PET, n = 8) and coincidence cameras (GC-PET, n = 7). Measurement of image quality was based on the NEMA (National Electrical Manufacturers Association) NU 2-2001 protocol and the IEC (International Electrotechnical Commission) body phantom. The latter contains six fillable spheres ranging in diameter from 37 mm down to 10 mm and a 'lung' insert. The two largest lesions L1-2 simulate cold lesions, the four smaller ones (L3-6) are filled with 18F and activity concentration ratios relative to background of 8:1 and 4:1, respectively. Acquisition and reconstruction in the study employed the participating institutes' standard oncological processing protocol. Calculation of contrast of the spheres was performed with a fully automated procedure. Contrast quality indices (CQIs) reflecting global performance were obtained by summing individual contrast values. Other image quality parameters calculated according to the NEMA protocol were background variability and relative error for correction of attenuation and scatter. Contrast values obtained were 61 +/- 16 and 37 +/- 14 for L1 (per cent contrast +/- SD for D-PET and GC-PET, respectively), 57 +/- 16 and 29 +/- 16 for L2, 46 +/- 10 and 26 +/- 6.3 for L3, 37 +/- 10 and 15 +/- 4.3 for L4, 26 +/- 11.5 and 6.1 +/- 2.5 for L5, 14 +/- 7.1 and 2.6 +/- 2.6 for L6, with D-PET systems consistently being superior to GC-PET systems. CQIs permitted ranking of the scanners, also demonstrating a clear distinction between D-PET and GC-PET systems. Background variability was largest for GC-PET systems; the relative error of attenuation and scatter correction was significantly correlated with image quality for D-PET systems only. The study demonstrated considerable differences in image quality not only between GC-PET and D-PET systems but also between individual D-PET systems with possible consequences for clinical interpretation of images and measurement of quantitative indices such as the standardized uptake value. The study provided valuable feedback to the participants as well as baseline data for improving interchangeability of PET images and of quantitative indices between different laboratories. PMID- 15876662 TI - A kinetic model for dynamic [18F]-Fmiso PET data to analyse tumour hypoxia. AB - A method is presented to identify and quantify hypoxia in human head-and-neck tumours based on dynamic [18F]-Fmiso PET patient data, using a model for the tracer transport. A compartmental model was developed, inspired by recent immunohistochemical investigations with the tracer pimonidazole. In order to take the trapping of the tracer and the diffusion in interstitial space into account, the kinetic model consists of two compartments and a specific input function. This voxel-based data analysis allows us to decompose the time-activity curves (TACs) into their perfusion, diffusion and hypoxia-induced retention components. This characterization ranges from well perfused tumours over diffusion limited hypoxia to strong hypoxia and necrosis. The overall shape of the TAC and the model parameters may point at the structural architecture of the tissue sample. The model addresses the two main problems associated with hypoxia imaging with PET. Firstly, the hypoxic areas are spatially separated from well perfused vessels, causing long diffusion times of the tracer. Secondly, tracer uptake occurs only in viable hypoxic cells, which constitute only a small subpopulation in the presence of necrosis. The resulting parameters such as the concentration of hypoxic cells and the perfusion are displayed in parameter plots ('hypoxia map'). Quantification of hypoxia performed with the presented kinetic model is more reliable than a criterion based on static standardized uptake values (SUV) at an early timepoint, because severely hypoxic/necrotic tissues show low uptake and are thus overlooked by SUV threshold identification. The derived independent measures for perfusion and hypoxia may provide a basis for individually adapted treatment planning. PMID- 15876663 TI - A prototype of a quasi-monochromatic system for mammography applications. AB - Improvement in image contrast and dose reduction, in mammographic x-ray imaging, can be achieved using narrow energy band x-ray beams in the 16-24 keV range. As part of an Italian Government funded project, a quasi-monochromatic system for mammography applications has been developed. The system is based on a tunable narrow energy band x-ray source operating in the 16-24 keV energy range. The bremsstrahlung beam is monochromatized via Bragg diffraction by a highly oriented pyrolytic graphite mosaic crystal (HOPG). The scanning system provides a large field (18 x 24 cm2) of quasi-monochromatic x-rays with energy resolution ranging from 10% at 18 keV to 17.2% at 24 keV. The system has been characterized in terms of fluence rate and energy resolution. An x-ray tube developed ad hoc allows us to acquire images in a reasonable time to minimize the motion blur. A qualitative analysis has been performed in order to know if the prototype system performances are far from a clinical application, by evaluating the spatial resolution, the field uniformity and the image quality as a function of the quasi-monochromatic beam energy. Dose evaluation has been performed as a function of the energy and compared to a conventional system for mammography. The quasi-monochromatic prototype system can produce comparable image quality at half the dose. PMID- 15876664 TI - Application of the compound probability density function for characterization of breast masses in ultrasound B scans. AB - The compound probability density function (pdf) is investigated for the ability of its parameters to classify masses in ultrasonic B scan breast images. Results of 198 images (29 malignant and 70 benign cases and two images per case) are reported and compared to the classification performance reported by us earlier in this journal. A new parameter, the speckle factor, calculated from the parameters of the compound pdf was explored to separate benign and malignant masses. The receiver operating characteristic curve for the parameter resulted in an A(z) value of 0.852. This parameter was combined with one of the parameters from our previous work, namely the ratio of the K distribution parameter at the site and away from the site. This combined parameter resulted in an A(z) value of 0.955. In conclusion, the parameters of the K distribution and the compound pdf may be useful in the classification of breast masses. These parameters can be calculated in an automated fashion. It should be possible to combine the results of the ultrasonic image analysis with those of traditional mammography, thereby increasing the accuracy of breast cancer diagnosis. PMID- 15876665 TI - Image reconstruction for the circle-and-arc trajectory. AB - Proposed is an exact shift-invariant filtered backprojection algorithm for the circle-and-arc trajectory. The algorithm has several important features. First, it allows for the circle to be incomplete. Second, axial truncation of the cone beam data is allowed. Third, the length of the arc is determined only by the region of interest and is independent of the size of the entire object. The algorithm is quite flexible and can be used for even more general trajectories that consist of several circular segments and arcs. The algorithm applies also in the case when the circle (or, circles) is complete. A numerical experiment with the clock phantom demonstrated good image quality. PMID- 15876666 TI - Extraction of dielectric properties of multiple populations from dielectrophoretic collection spectrum data. AB - In this paper we show how simplifying assumptions can be used to extract useful data from the dielectrophoretic collection spectrum, in particular for the cytoplasm, and hence determine the properties of multiple populations of cells within a sample. Specifically, the observation of the frequencies of onset of dielectric dispersion allows the identification and enumeration of populations of cells according to cytoplasmic conductivity, with particular relevance to the determination of the action of drugs for high-throughput screening applications. PMID- 15876667 TI - Responses of a diamond detector to high-LET charged particles. AB - The responses of a commercial diamond detector (type 60003, PTW-Freiburg) to several heavy ions were examined. The responses to heavy-ion beams reached stable levels with relatively small pre-irradiation doses compared to photon-beam irradiations. The responses also reached stable levels with a smaller pre irradiation dose when the dose rate of the He beams was increased. A total accumulated dose of about 5 Gy was required for the pre-irradiation dose of heavy ion beams. No angular dependence of the detector responses was observed within a deviation of 5%. The dose-rate dependence of the detector responses to heavy-ion beams was far smaller than that to gamma rays. The decrease in the response was within 0.9%, with a variation from 0.88 to 18.2 Gy min(-1) in the carbon beam. We examined the LET dependence of the diamond detector responses using various kinds of heavy-ion beams. The responses had particle dependence in addition to LET dependence. The responses decreased more with higher LET particles and decreased less with large-Z particles. We proposed a gradual-saturation model based on the track structure under several simple assumptions to explain the LET and particle dependences of the response. PMID- 15876669 TI - Localization and quantification of fluorescent inclusions embedded in a turbid medium. AB - A set-up, based on a CCD camera, to localize fluorescent inclusions in diffusing media was developed. This set-up allows one to acquire a huge dataset along two axes. This aspect is fundamental to performing a tomographic reconstruction in order to quantify the fluorescence amplitude in each voxel of the sample. Firstly, a simple analytical approach to recover the position of a single inclusion, embedded in a turbid medium, was developed. Then, we implemented a reconstruction algorithm to recover the position of one and two inclusions and to estimate their relative concentrations. Finally, we studied the dependence of reconstructed data on the number of injection points of excitation light and the number of detection points of fluorescence emission. PMID- 15876668 TI - A method for determination of the absorption and scattering properties interstitially in turbid media. AB - We have developed a method to quickly determine tissue optical properties (absorption coefficient mu(a) and transport scattering coefficient mu'(s)) by measuring the ratio of light fluence rate to source power along a linear channel at a fixed distance (5 mm) from an isotropic point source. Diffuse light is collected by an isotropic detector whose position is determined by a computer controlled step motor, with a positioning accuracy of better than 0.1 mm. The system automatically records and plots the light fluence rate per unit source power as a function of position. The result is fitted with a diffusion equation to determine mu(a) and mu'(s). We use an integrating sphere to calibrate each source-detector pair, thus reducing uncertainty of individual calibrations. To test the ability of this algorithm to accurately recover the optical properties of the tissue, we made measurements in tissue simulating phantoms consisting of Liposyn at concentrations of 0.23, 0.53 and 1.14% (mu'(s) = 1.7-9.1 cm(-1)) in the presence of Higgins black India ink at concentrations of 0.002, 0.012 and 0.023% (mu(a) = 0.1-1 cm(-1)). For comparison, the optical properties of each phantom are determined independently using broad-beam illumination. We find that mu(a) and mu'(s) can be determined by this method with a standard (maximum) deviation of 8% (15%) and 18% (32%) for mu(a) and mu'(s), respectively. The current method is effective for samples whose optical properties satisfy the requirement of the diffusion approximation. The error caused by the air cavity introduced by the catheter is small, except when mu(a) is large (mu(a) > 1 cm( 1)). We presented in vivo data measured in human prostate using this method. PMID- 15876670 TI - An FDK-like cone-beam SPECT reconstruction algorithm for non-uniform attenuated projections acquired using a circular trajectory. AB - In this paper, Novikov's inversion formula of the attenuated two-dimensional (2D) Radon transform is applied to the reconstruction of attenuated fan-beam projections acquired with equal detector spacing and of attenuated cone-beam projections acquired with a flat planar detector and circular trajectory. The derivation of the fan-beam algorithm is obtained by transformation from parallel beam coordinates to fan-beam coordinates. The cone-beam reconstruction algorithm is an extension of the fan-beam reconstruction algorithm using Feldkamp-Davis Kress's (FDK) method. Computer simulations indicate that the algorithm is efficient and is accurate in reconstructing slices close to the central slice of the cone-beam orbit plane. When the attenuation map is set to zero the implementation is equivalent to the FDK method. Reconstructed images are also shown for noise corrupted projections. PMID- 15876671 TI - Verification of the optimal backscatter for an aSi electronic portal imaging device. AB - Electronic portal imaging devices (EPIDs), currently used for determining proper patient placement during irradiation in a radiotherapy treatment, can also be used as dosimeters. However, the Varian aS500 portal imager exhibits dosimetric artefacts caused by non-uniform backscatter from mechanical support structures located behind the imager. Monte Carlo simulations predict that adding 5 mm of Pb behind the imaging cassette will reduce the non-uniform backscatter to <1% for 6 MV and to <1.5% for 18 MV photon beams. This study experimentally tested this hypothesis by comparing images using an unmodified test imager and an imager modified by adding 3 and 5 mm of Pb behind the imaging cassette. Using the modified imager containing 5 mm of Pb, the non-uniform backscatter was reduced to <0.5% for 6 MV and <0.6% for 18 MV beams. Addition of the 5 mm of Pb increased the detector contrast by 3.5% +/- 0.5% at 6 MV and 5.0% +/- 0.7% at 18 MV, and increased the resolution by 0.9% +/- 0.2% at 6 MV and 0.5% +/- 0.12% at 18 MV. PMID- 15876672 TI - Multiple projection optical diffusion tomography with plane wave illumination. AB - We describe a new data collection scheme for optical diffusion tomography in which plane wave illumination is combined with multiple projections in the slab imaging geometry. Multiple projection measurements are performed by rotating the slab around the sample. The advantage of the proposed method is that the measured data are more compatible with the dynamic range of most commonly used detectors. At the same time, multiple projections improve image quality by mutually interchanging the depth and transverse directions, and the scanned (detection) and integrated (illumination) surfaces. Inversion methods are derived for image reconstructions with extremely large data sets. Numerical simulations are performed for fixed and rotated slabs. PMID- 15876673 TI - Gauss-Newton method for image reconstruction in diffuse optical tomography. AB - We present a regularized Gauss-Newton method for solving the inverse problem of parameter reconstruction from boundary data in frequency-domain diffuse optical tomography. To avoid the explicit formation and inversion of the Hessian which is often prohibitively expensive in terms of memory resources and runtime for large scale problems, we propose to solve the normal equation at each Newton step by means of an iterative Krylov method, which accesses the Hessian only in the form of matrix-vector products. This allows us to represent the Hessian implicitly by the Jacobian and regularization term. Further we introduce transformation strategies for data and parameter space to improve the reconstruction performance. We present simultaneous reconstructions of absorption and scattering distributions using this method for a simulated test case and experimental phantom data. PMID- 15876674 TI - Investigation of dose homogeneity for loose helical tomotherapy delivery in the context of breath-hold radiation therapy. AB - Loose helical delivery is a potential solution to account for respiration-driven tumour motion in helical tomotherapy (HT). In this approach, a treatment is divided into a set of interlaced 'loose' helices commencing at different gantry angles. Each loose helix covers the entire target length in one gantry rotation during a single breath-hold. The dosimetric characteristics of loose helical delivery were investigated by delivering a 6 MV photon beam in a HT-like manner. Multiple scenarios of conventional 'tight' HT and loose helical deliveries were modelled in treatment planning software, and carried out experimentally with Kodak EDR2 film. The advantage of loose helical delivery lies in its ability to produce a more homogeneous dose distribution by eliminating the 'thread' effect an inherent characteristic of HT, which results in dose modulations away from the axis of gantry rotation. However, loose helical delivery was also subjected to undesirable dose modulations in the direction of couch motion (termed 'beating' effect), when the ratio between the number of beam projections per gantry rotation (n) and pitch factor (p) was a non-integer. The magnitude of dose modulations decreased with an increasing n/p ratio. The results suggest that for the current HT unit (n = 51), dose modulations could be kept under 5% by selecting a pitch factor smaller than 7. A pitch factor of this magnitude should be able to treat a target up to 30 cm in length. Loose helical delivery should increase the total session time only by a factor of 2, while the planning time should stay the same since the total number of beam projections remains unchanged. Considering its dosimetric advantage and clinical practicality, loose helical delivery is a promising solution for the future HT treatments of respiration-driven targets. PMID- 15876675 TI - Compensation for respiratory motion by gated radiotherapy: an experimental study. AB - Respiratory organ motion is known to be one of the largest intrafractional organ motions. Therefore, it is important to investigate the potential benefit of gated dose delivery approaches which aim to account for the respective dose uncertainties. In this study respiration is simulated by a moving lung phantom; the movement is not restricted to a normal sinusoidal progression and simulates the one of the embedded lung tumour in the cranial-caudal direction. An IMRT plan with a total of 29 beam segments was designed for the treatment of this tumour. It was irradiated in its resting position-which is the position at exhalation-and during movement. Furthermore the irradiation was triggered using different amplitude thresholds, which means that the irradiation only proceeded if the deviation of the tumour's position from its resting position is smaller than the given threshold. We determined the gating-related increase of the treatment time for various gating procedures. We also measured the resulting dose distribution in specific slices of the phantom perpendicular to the direction of the movement using film dosimetry and compared it to the dose distribution of the static case. Since these film measurements cannot be done inside the whole tumour, additionally the movement and gating was simulated using the planning software to calculate the 3D dose distribution inside the tumour and to generate dose volume histograms for different treatment modalities. The total treatment time was observed to increase by 20%-100% depending on the individual gating threshold and can be calculated easily. The analysis of the films showed that irradiation without gating leads to significant underdosages up to 33%, especially at the edge of the tumour. With gating it is possible to considerably reduce this underdosage down to 9% depending on the trigger threshold. The calculation of the dose volume histograms makes it possible to find a reasonable compromise between the improvement of the dose distribution and the increase of the treatment time. PMID- 15876676 TI - A novel approach for the averaging of magnetocardiographically recorded heart beats. AB - Performing signal averaging in an efficient and correct way is indispensable since it is a prerequisite for a broad variety of magnetocardiographic (MCG) analysis methods. One of the most common procedures for performing the signal averaging to increase the signal-to-noise ratio (SNR) in magnetocardiography, as well as in electrocardiography (ECG), is done by means of spatial or temporal techniques. In this paper, an improvement of the temporal averaging method is presented. In order to obtain an accurate signal detection, temporal alignment methods and objective classification criteria are developed. The processing technique based on hierarchical clustering is introduced to take into account the non-stationarity of the noise and, to some extent, the biological variability of the signals reaching the optimum SNR. The method implemented is especially designed to run fast and does not require any interaction from the operator. The averaging procedure described in this work is applied to the averaging of MCG data as an example, but with its intrinsic properties it can also be applied to the averaging of ECG recording, averaging of body-surface-potential mapping (BSPM) and averaging of magnetoencephalographic (MEG) or electroencephalographic (EEG) signals. PMID- 15876677 TI - The modified linear-quadratic model of Guerrero and Li can be derived from a mechanistic basis and exhibits linear-quadratic-linear behaviour. PMID- 15876678 TI - Mapping QTLs for alpha-amylase activity in rye grain. AB - Genetic control of alpha-amylase activity in rye grain was investigated by QTL mapping based on DS2 x RXL10 intercross consisting of 99 F5-6 families propagated at one location during four vegetation seasons. A wide range of variation in alpha-amylase activity and transgression effects were found among families and parental lines. This variation was shown to be determined in 40.1% by 7 significant (LOD score not less than 2.5) and 2 putative QTLs (2 < LOD < 2.5) distributed on all rye chromosomes except 4R. Two significant QTLs located on 3RL and 5RL chromosome arms were expressed each year. The third significant QTL was detected in three years (1RL). The other four significant QTLs (2RL, 5RS, 6RL, 7RL) were found in one year of study. The number and composition of QTLs were specific for a given year varying from three to six. QTLs were not correlated with isoenzyme polymorphisms at the structural alpha-Amy1 loci. A QTL associated with a region containing the alpha-Amy3 locus was detected on chromosome 5RL. Both high- and low-activity QTL alleles were found in each parental line, which explains the appearance of transgressive recombinants in the segregating population. PMID- 15876679 TI - Assessing genetic variation to predict the breeding value of winter triticale cultivars and lines. AB - The objective of this study was to evaluate the usefulness of amplified fragment length polymorphism (AFLP) analyses for selection of the best parents for breeding of hybrid winter triticale. Phenotypic diversity was measured for 8 agronomic traits in 10 parents and 27 F1 hybrids. Genotypic diversity was measured by 91 AFLP markers. Coefficients of correlation of genetic similarity (AFLP-GS) with both Euclidean distances and mean values of the traits were generally not significant. A correction of the preliminary binary matrix into trait-specific derived matrices increased the values of 5 correlation coefficients to a significant level. The correlation of AFLP-GS with mid-parent heterosis of grain weight per plant was low but significant (r = -0.452). Our study confirms the effectiveness of marker preselection for obtaining AFLP-GS better correlated with heterosis. The use of derived matrices is promising for reducing the number of cross combinations tested for specific combining ability. PMID- 15876680 TI - Inheritance of seed dormancy in Tibetan semi-wild wheat accession Q1028. AB - Tibetan semi-wild wheat (Triticum aestivum ssp. tibetanum Shao) is one of the Chinese endemic hexaploid wheat genetic resources, distributed only in the Qinghai-Xizang Plateau of China. It has special characters, such as a hulled glume and spike disarticulation. However, seed dormancy, another important character for wheat resistance to pre-harvest sprouting, was rarely reported. Seed dormancy of more than 10 Tibetan semi-wild wheat accessions was evaluated, and their germinations were 0% or near 0% with both treatments of threshed seeds and intact spikes at hard dough stage. Tibetan semi-wild wheat accession Q1028 was investigated for its seed dormant characters by testing the seed germination percentages of intact spikes, seeds with bract powder, normal seeds, seeds with pierced coat, and sectioned embryos. It was observed that embryo dormancy of Q1028 accounted for its seed dormancy. Using threshed seeds and intact spikes at hard dough stage, the inheritance of seed dormancy was carried out using the F1, F2, F3 and F2BC1 populations of the cross between Q1028 and a wheat line 88-1643, susceptible to preharvest sprouting. The germinations of seeds and intact spikes in F1 plants were 1.0% and 0.9%, respectively. It indicated that seed dormancy of Q1028 was inherited as a dominant trait. From the genetic analysis of the F2, F3 and F2BC1 populations it was found that the strong seed dormancy of Q1028 was controlled by two dominant genes. PMID- 15876681 TI - Efficiency of transformation of Polish cultivars of pea (Pisum sativum L.) with various regeneration capacity by using hypervirulent Agrobacterium tumefaciens strains. AB - An Agrobacterium-mediated transformation method of pea has been developed for several edible and fodder cultivars of pea (Pisum sativum L.), characterized previously in their potential for regeneration via organogenesis. The most appropriate explant, which was susceptible to Agrobacterium infection and capable of regenerating transgenic plants, turned out to be a slice of an immature embryo, including the embryo axis and the basal part of a cotyledon. Three hypervirulent strains of A. tumefaciens were tested: AgL0, AgL1 and EHA105. Each carried the binary vector pP35SGIB containing the uid gene, with an intron under control of the 35S promoter, and the bar gene conferring resistance to phosphinotricin. Strain AgL0 was found to be efficient for the majority of cultivars, followed by AgL1 and EHA105. Transformation efficiency varied from 0.7 to 4.1%, depending on cultivar and Agrobacterium strain. The transformation efficiency of particular pea cultivars did not clearly correspond to their regeneration capacity, which--although indispensable--was not a critical parameter of successful transformation. The presence of integrated genes in pea genomic DNA was detected by the PCR. T-DNA was stably transmitted to the progeny, as it was confirmed by Southern hybridization. The activity of introduced genes was analysed by the histochemical GUS assay and by painting leaves or by spraying transgenic plants with the herbicide Basta. PMID- 15876682 TI - Production and characterization of amphiploids of Aegilops kotschyi and Ae. biuncialis with Secale cereale, and of backcross hybrids of Ae. biuncialis x S. cereale amphiploids with 2x and 4x S. cereale. AB - Amphiploids (2n = 6x = 42) of Ae. kotschyi and Ae. biuncialis with self compatible S. cereale were produced from F1 sterile hybrids (2n = 3x = 21) through colchicine treatment and callus tissue regeneration. The amphiploids resembled the F1 plants in overall morphology, but were larger in all respects and self-fertile. The spikelets consisted mostly of 3 well-developed florets. Selfed seeds were obtained from some colchicine-doubled sectors and callus regenerates. Most of the produced seeds were well developed. Backcrosses between amphiploids and rye (2x and 4x) resulted in obtaining (Ae. biuncialis x S. cereale amphiploid) x S. cereale hybrids via embryo culture. The BC1 plants (2n = 4x = 28 and 2n = 5x = 35, respectively) were phenotypically intermediate between the parents and vigorous in vegetative growth. Some seeds were obtained only from the 35-chromosome BC1 hybrids. PMID- 15876683 TI - Analysis of oocyte quality in recombinant inbred mouse strains developed from KE and CBA strains that differ in fertilization efficiency. AB - Recombinant inbred (RI) mouse strains were developed from reciprocal crosses between two inbred strains differing in the proportion of fertilized ova (CBA, 100%; KE, 77%), to analyse the underlying factors. A correlation (r = 0.83, P < 0.01) between fertilization efficiency within 22 RI strains and after mating RI females with KE males proved that oocyte quality was involved. The following oocyte parameters were analysed in RI and progenitor strains: time of meiotic maturation, rapidity of enzymatic removal of egg investments, and proportion of fertilized ova with supplementary spermatozoa in the perivitelline space. Among the RI strains, high incidence of supplementary spermatozoa was correlated with lower efficiency of fertilization (r = -0.58, P < 0.05) and with slow meiotic maturation (r = -64, P < 0.01), suggesting that delayed maturation may affect oocyte ability of being fertilized by the first penetrating spermatozoon. However, significant correlations were also found between characters which coexist within the progenitor strains, but are not likely to be physiologically related; this suggests that RI strains have inherited large blocks of progenitor genomes, not disrupted by recombination. The strain distribution pattern (SDP) of the analysed traits revealed CBA-like, KE-like, and intermediate phenotypes, indicating that they are polygenically determined. No linkages were found between the studied traits and 12 enzymatic markers. However, the SDP for fertilization efficiency showed a preponderance of non-matching strains (15/19) in relation to agouti locus; the known instability of this chromosome region makes it possible that a putative linkage was disrupted by recombination when RI strains were created. PMID- 15876684 TI - Immunorelevant gene expression in LPS-challenged bovine mammary epithelial cells. AB - Infection of the bovine mammary gland, in addition to causing animal distress, is a major economic burden of the dairy industry. Greater understanding of the initial host response to infection may lead to more accurate selection of resistant animals or to novel prophylactic or therapeutic intervention strategies. The epithelial cell plays a role in the host response by alerting the immune system to the infection and providing a signal as to where the infection is located. To understand this process better, a cDNA microarray approach was used to search for potential signals produced by mammary epithelial cells in response to exposure to Escherichia coli lipopolysaccharide (LPS). Total RNA from separate cultures of epithelial cells from 4 Holstein cows was harvested 6 h after LPS challenge or control conditions. For each cow, RNA from control or LPS exposed cells was transcribed to cDNA and labeled with Cy3 or Cy5, then pooled and applied to a bovine total leukocyte (BOTL) microarray slide containing 1278 unique transcripts. Dye reversal was used so that RNA from two of the control cultures was labeled with Cy3 while RNA from the other two control cultures was labeled with Cy5. From the resulting microarray data we selected 4 of the 9 genes significantly (P < 0.02) induced (>1.25-fold) in response to LPS exposure for more detailed analysis. The array signal intensity for 3 of these genes, RANTES/CCL5, IL-6 and T-PA, was relatively low, but quantitative real-time RT-PCR (Q-RT-PCR) analysis revealed that they were induced 208-fold, 10-fold and 3-fold, respectively. The gene that showed the greatest fold induction by microarray analysis (2.5-fold) was CXCL5. This gene had a relatively strong signal intensity on the array and was easily detected by northern blot analysis, which indicated a 10-fold induction. This cell culture model system provides evidence for an important role of the mammary epithelial cell in initiating the innate response to infection. PMID- 15876685 TI - Nucleotide sequence polymorphism within exon 4 of the bovine prolactin gene and its associations with milk performance traits. AB - Prolactin plays an important regulatory function in mammary gland development, milk secretion, and expression of milk protein genes. Hence the PRL gene is a potential quantitative trait locus and genetic marker of production traits in dairy cattle. We analysed the sequence of the PRL gene to investigate whether mutations in this sequence might be responsible for quantitative variations in milk yield and composition. Using SSCP and direct sequencing, we detected six single-nucleotide polymorphisms within a 294-bp prolactin gene fragment involving exon 4. All detected mutations were silent with respect to the amino acid sequence of the protein. PCR-RFLP genotyping of SNP 8398 R (RsaI) was used to assess allele frequencies in 186 Black-and-White cows (0.113 and 0.887 for A and G, respectively) and in 138 Jersey cows (0.706 and 0.294 for A and G, respectively). Black-and-White cows with genotype AG showed the highest milk yield, while cows with genotype GG showed the highest fat content. PMID- 15876686 TI - Inbreeding and relationship in Polish Black-and-White sires. AB - The aim of the study was to analyse inbreeding and relationship in the Polish population of Black-and-White sires. Data were pedigrees of 25,036 Black-and White sires born from 1960 through 2000, divided into subsets of 11,447 proven and 13 589 unproven sires, and their 38,228 ancestors, altogether 63,264 animals. Average inbreeding coefficients were about 0.3% for both subsets of sires. Mean relationship coefficients ranged from 0.1% in all animals to 0.7% in the subset of proven sires. Positive time trends in inbreeding coefficients were observed when the subsets of sires were divided into 5-year intervals according to the year of birth and in terms of Holstein Friesian gene contribution. PMID- 15876687 TI - Association of PIT1, GH and GHRH polymorphisms with performance and carcass traits in Landrace pigs. AB - The study of candidate genes, based on physiological effects, is an important tool to identify genes to be used in marker-assisted selection programs. In this study, a group of halothane gene-free, non-castrated, male Landrace pigs was used to study the association between polymorphisms in the PIT1 (n = 218), GH (n = 213) and GHRH (n = 206) genes and fat thickness, average daily gain, and the EPD (expected progeny difference) for fat thickness, average daily gain, and litter size. These genes are potential candidate markers because of their important physiological effects. The pigs were genotyped by PCR-RFLP, and the statistical model used to analyze the association between genotypes and the traits measured included genotypes as a fixed effect and age and weight as covariates. PIT1 polymorphisms were associated with fat thickness (P = 0.0019), EPD for average daily gain (P = 0.0001) and EPD for fat thickness (P = 0.0001), whereas GH polymorphisms were associated with fat thickness (P = 0.0326) and average daily gain (P = 0.0127), and GHRH polymorphisms were associated with the average daily gain (P = 0.0001) and EPD for fat thickness (P = 0.0004). These results confirmed the potential usefulness of these genes in marker-assisted selection programs for pig breeding. PMID- 15876688 TI - Sequence polymorphism in a novel noncoding region of Pacific oyster mitochondrial DNA. AB - Nucleotide sequence polymorphism in a 641-bp novel major noncoding region of mitochondrial DNA (mtDNA-NC) of the Pacific oyster Crassostrea gigas was analysed for 29 cultured individuals within the Goseong population. A total of 30 variable sites were detected, and the relative frequency of nucleotide alteration was determined to be 4.68. Alterations were mostly single nucleotide substitutions. Transition, transversion, both transition and transversion, and both transversion and nucleotide deletion were observed at 18, 9, 2 and 1 sites, respectively. Among 29 specimens, 22 haplotypes were identified, and pairwise genetic diversity of haplotypes was calculated to be 0.988 from multiple sequence substitutions using the two-parameter model. A phylogenetic tree, obtained for haplotypes by the neighbor-joining method, showed a single cluster of linkages. The cluster comprised 11 haplotypes associating with 14 specimens, while the other 11 haplotypes associating with 15 specimens were scattered. This mtDNA-NC presenting a high nucleotide sequence polymorphism is a potential mtDNA control region. It therefore can serve as a genetic marker for intraspecies phylogenetic analysis of the Pacific oyster and is more useful than the less polymorphic mtDNA coding genes. PMID- 15876689 TI - Molecular methods for rapid detection of aneuploidy. AB - Rapid molecular biological methods for prenatal diagnosis of the most common aneuploidies, collectively known as rapid aneuploidy testing, are compared in this review. We discuss methodological problems and limitations of these various methods. All these techniques are believed to be accurate and carry a low risk of misdiagnosis, but they differ in terms of labour-intensity and amenability to automation and high throughput testing. The question how to apply them safely and economically in a clinical setting has not been answered yet. The discussed techniques are so far not used as stand-alone tests, but some of them are routinely applied as a preliminary test that shortens the waiting time for classic cytogenetic karyotyping. In the future, mainly because of economical reasons, these methods may replace cytogenetics in the category of patients who make up the majority of those currently offered prenatal karyotyping: patients with moderately increased risk and no abnormalities detected by ultrasound. PMID- 15876690 TI - DNA constructs designed to produce short hairpin, interfering RNAs in transgenic mice sometimes show early lethality and an interferon response. AB - Arylamine N-acetyltransferase (NAT) genes were targeted for inhibition using short hairpin RNA (shRNA) using two different RNA polymerase III promoters. Constructs were developed for NAT1 and NAT2, the endogenous mouse genes, and for human NAT1. There were fetal and neonatal deaths with these constructs, perhaps due in part to an interferon response as reflected in increases in oligoadenylate synthetase I mRNA levels. Seven out of 8 founders with the U6 promoter generated offspring but only 2 gave positive offspring. Out of 15 founders for H1 promoted constructs, only 4 had positive offspring. When transgenic lines were successfully established, the expression of the targeted genes was variable between animals and was not generally inhibitory. PMID- 15876691 TI - Myosin heavy chain protein and gene expression in the masseter muscle of adult patients with distal or mesial malocclusion. AB - The aim of this study was to determine the amount of myosin heavy chain (MyHC) proteins and MyHC mRNA in muscles of patients with different positions of the mandible. Ten adult patients for orthognathic surgery were divided into two groups: distal and mesial malocclusion. The mRNA expression of two MyHC isoforms of the anterior and posterior part of the right and left side of the human masseter muscle was analysed with a competitive RT-PCR assay. An exogenous template that includes oligonucleotide sequences specific for sarcomeric MyHC isoforms (1 and 2x) was constructed and utilized as competitor. Different isoforms of the MyHC protein were identified by Western blot analysis. In the total mRNA pool of the masseter muscle, the MyHC 1 mRNA level was 25.5 +/- 7.6% and the MyHC 2x mRNA was 2.5 +/- 1.2%. The anterior part of the masseter muscle from patients with distal occlusion contained more type 1 and 2x MyHC mRNA, as compared to patients with mesial occlusion (P < 0.05). No difference in the protein distribution was observed. The differences in mRNA expression may be caused by the enforced stress of the masticatory muscle in distal occlusion because of the disadvantageous pivot. PMID- 15876692 TI - CAG repeat polymorphism in the androgen receptor (AR) gene of SBMA patients and a control group. AB - Spinobulbar muscular atrophy (SBMA) is an X-linked form of motor neuron disease characterized by progressive atrophy of the muscles, dysphagia, dysarthria and mild androgen insensitivity. SBMA is caused by CAG repeat expansion in the androgen receptor gene. CAG repeat polymorphism was analysed in a Polish control group (n = 150) and patients suspected of SBMA (n = 60). Normal and abnormal ranges of CAG repeats were established in the control group and in 21 patients whose clinical diagnosis of SBMA was molecularly confirmed. The ranges are similar to those reported for other populations. PMID- 15876693 TI - Optimisation of the microbiological mutagenicity assay based on genetically modified Vibrio harveyi strains. AB - Recently, we have developed a novel assay designed for detection of mutagenic pollution of the marine environment. This assay is based on the use of a series of genetically modified strains (named BB7, BB7M, BB7X and BB7XM) of a marine bacterium Vibrio harveyi. Sensitivity of the V. harveyi mutagenicity assay was found to be similar to, or even somewhat higher than, that of the commonly used Ames test. Subsequent studies indicated that this assay may be useful in assessment of mutagenic contamination of the marine environment. Nevertheless, we assumed that improvement of this assay is still possible, and thus we aimed to optimise its procedures. Here we present our research on the optimisation of the V. harveyi mutagenicity assay, which indicated that different tester strains used in this assay give the best results depending upon the experimental conditions employed. Incubation of bacteria in a buffer, rather than in a nutrient broth, containing a mutagen, increased the efficiency of the assay with BB7 and BB7M strains, but had a deleterious effect in the case of BB7X and BB7XM. The latter couple of strains revealed higher mutagenicity in the plate assay, as compared to the liquid medium assay. However, the opposite effect was observed for BB7 and BB7M. Low-dose (1 J m(-2)) UV irradiation, as well as 30 min incubation in 0.1 M CaCl2, had no significant effect on the efficiency of the assay when using BB7 and BB7M, whereas the number of mutagen-induced mutants of BB7X and BB7XM strains increased about two times under these conditions. Our previous experiments indicated that various tester strains revealed different sensitivity to particular mutagens. Thus, a series of strains should be used in the assay. Results presented in this report show that different conditions should be used for two pairs of the tester strains: BB7 and BB7M, and BB7X and BB7XM. PMID- 15876694 TI - Ag3.5Bi7.5S13, a new member (N = 8) of the homologous series [Bi2S3]2.AgBiS2]((N 1)/2). AB - The silver bismuth tridecasulfide Ag3.5Bi7.5S13 crystallizes in the monoclinic space group C2/m. Its structure is built up of two alternating kinds of layered modules parallel to (001). In the module denoted A, octahedra around the metal positions (M = Ag/Bi, M2 and an S atom on 2/m, other atoms on m) alternate with paired monocapped trigonal prisms around Bi. The NaCl-type module B is composed of parallel eight-membered chains of edge-sharing octahedra running diagonally across it. Ag3.5Bi7.5S13 is the member with N = 8 of the pavonite homologous series (N)P of ternary compounds with the general formula [Bi2S3]2.[AgBiS2]((N 1)/2). PMID- 15876695 TI - RbGa3(P3O10)2: a new gallium phosphate isotypic with RbAl3(P3O10)2. AB - Rubidium trigallium bis(triphosphate), RbGa3(P3O10)2 has been synthesized by solid-state reaction and studied by single-crystal X-ray diffraction at room temperature. This compound is the first anhydrous gallium phosphate containing both GaO4 tetrahedra (Ga1) and GaO6 octahedra (Ga2 and Ga3). The three independent Ga atoms are located on sites with imposed symmetry 2 (Wickoff positions 4a for Ga1 and 4b for Ga2 and Ga3). The GaO4 and GaO6 polyhedra are connected through the apices to triphosphate groups and form a three-dimensional host lattice. This framework presents intersecting tunnels running along the [001] and <110> directions, where the Rb2+ cations are located on sites with imposed symmetry 2 (Wickoff position 4a). The structure also exhibits remarkable features, such as infinite helical columns created by the junction of GaO4 and PO4 tetrahedra. PMID- 15876696 TI - The twofold superstructure of titanium(III) oxybromide at T = 17.5 K. AB - The low-temperature (T = 17.5 K) structure of titanium(III) oxybromide, TiOBr, is reported as a twofold superstructure of the crystal structure at room temperature. Weak superlattice reflections were measured with synchrotron radiation X-rays and were analyzed by structure refinements employing superspace techniques. Both the low-temperature and the room-temperature structures of TiOBr are isostructural with the corresponding structures of TiOCl. The results indicate that at low temperatures TiOBr is in a spin-Peierls state, similar to that of TiOCl, but with the modulations and relevant interactions smaller than in the latter compound. PMID- 15876697 TI - Barium chlorite hydrate, Ba(ClO2)2.3.5H2O. AB - The structure of barium chlorite hydrate, Ba(ClO2)2.3.5H2O, has been determined by single-crystal X-ray analysis at 150 K. The structure is monoclinic, space group C2/c, with Z = 8. It contains layers of Ba2+ cations coordinated by ClO2- anions and water molecules. There are also solvate water molecules involved only in hydrogen bonding of the layers. Three solvate water O atoms are on sites of twofold symmetry, while all other atoms are in general positions. The full coordination environment of the Ba2+ cation consists of ten O atoms belonging to six chlorites and three water molecules, forming a bicapped square antiprism. PMID- 15876698 TI - Two cubic polymorphs of AlGeLi. AB - Aluminium germanium lithium, AlGeLi, crystallizes in two cubic dimorphs. The structure of the F-43m form, already inferred from powder data, has been confirmed by both powder and single-crystal X-ray diffraction studies. The second dimorph, not previously identified, adopts a disordered centrosymmetric structure with space group Fm-3m. PMID- 15876699 TI - A new strontium borophosphate, Sr6BP5O20, from synchrotron powder data. AB - Strontium borophosphate, Sr6BP5O20, was prepared by a solution synthesis method. The crystal structure was solved ab initio from synchrotron powder data without preliminary knowledge of the chemical formula. The compound crystallizes in space group -4c2. Sr atoms occupy sites coordinated by eight or nine O atoms, and the anionic layer consists of BO4 and PO4 tetrahedra. The eightfold-coordinated Sr atom lies at a site with twofold symmetry, while one P atom and the B atom are located on special positions of site symmetry -4. PMID- 15876700 TI - [Li(MoO2)2O(AsO4)]. AB - Single crystals of the molybdenum arsenate Li(MoO2)2O(AsO4) [lithium bis(dioxomolybdenum) arsenate oxide] have been prepared by solid-state reaction. The framework of Li(MoO2)2O(AsO4) consists of corner-shared Mo2AsO12 structural units, which form layers parallel to the ab plane. The Mo2AsO12 units result from the association of corner-shared AsO4 tetrahedra and Mo2O9 dimers. The Li+ ions are located between layers in a square-based-pyramidal environment. This structure is compared with those containing M2O9 dimers (M = Mo, Fe or V). The As, Li and one of the O atoms lie at sites with mm symmetry, while the Mo atom and two of the O atoms lie at sites with m symmetry. PMID- 15876701 TI - Hexaaquanickel(II) bis(4',7-dimethoxyisoflavone-3'-sulfonate) octahydrate. AB - In the title compound, [Ni(H2O)6](C17H13O7S)2.8H2O, the NiII atom is located on an inversion centre in the space group P2(1)/c. The [Ni(H2O)6]2+, C17H13O7S- and H2O components form many hydrogen bonds and there are pi-pi stacking interactions between the isoflavone units. The hydrogen bonds, pi-pi stacking interactions and electrostatic interactions between the cation and anions link the components into a three-dimensional structure. PMID- 15876702 TI - catena-Poly[[[diaquabis(pyridine-4-carboxamide-kappaN)copper(II)]-mu2-squarato kappaO1:O3] dihydrate]. AB - The asymmetric unit of the title compound, {[Cu(C4O4)(C6H6N2O)2(H2O)2].2H2O}n, consists of one pyridine-4-carboxamide (isonicotinamide or ina) ligand, one-half of a squarate dianion, a coordinated aqua ligand and a solvent water molecule. Both the CuII and the squarate ions are located on inversion centers. The CuII ions are octahedrally surrounded by four O atoms of two water molecules and two squarate anions, and by two N atoms of the isonicotinamide ligands. The crystal structure contains chains of squarate-1,3-bridged CuII ions. These chains are held together by N-H...O and O-H...O intermolecular hydrogen-bond interactions, forming an extensive three-dimensional network. PMID- 15876703 TI - Bis(1,3-diphenylpropane-1,3-dionato-kappa2O,O'){hydrotris[3-(2-pyridyl)pyrazol-1 yl]borato}praseodymium(III): another member of an unpredictable series. AB - Reaction of praseodymium(III) chloride with stoichometric quantities of dibenzoylmethane (Hdbm) and hydrotris[3-(2-pyridyl)pyrazol-1-yl]borate (Tp 2py) affords the title complex, [Pr(C24H19BN9)(C15H11O4)2]. The lanthanide ion in this ternary complex exhibits an N6O4 ten-coordinate geometry arising from the five bidentate arms found on the anionic ligands. The structure is entirely different from those found in other lanthanide complexes with the same ligand set. PMID- 15876704 TI - catena-Poly[[[N-(4-amino-1,6-dihydro-1-methyl-5-nitroso-6-oxopyrimidin-2-yl)-(S) glutamato]hexaaquabarium]-mu-N-(4-amino-1,6-dihydro-1-methyl-5-nitroso-6 oxopyrimidin-2-yl)-(S)-glutamato]: coordination polymer chains linked into a three-dimensional framework by N-H...O and O-H...O hydrogen bonds. AB - In the title complex, [Ba(C10H12N5O6)2(H2O)6]n, the Ba atom is nine-coordinated by six water ligands and three carboxylate O atoms. The Ba2+ cations and the anionic glutamate ligands form coordination polymer chains, and these chains are linked by pairs of N-H...O hydrogen bonds and pairs of O-H...O hydrogen bonds to form a continuous three-dimensional framework of cations and anions, which is reinforced by hydrogen bonds involving the water molecules. PMID- 15876705 TI - Bis(acesulfamato-kappa2N3,O4)bis(2-aminopyrimidine-kappaN1)copper(II). AB - In the crystal structure of the title compound, bis(2-aminopyrimidine kappaN1)bis[6-methyl-1,2,3-oxathiazin-4(3H)-one 2,2-dioxide(1-) kappa2N3,O4]copper(II), [Cu(C4H4NO4S)2(C4H5N3)2], the first mixed-ligand complex of acesulfame, the CuII centre resides on a centre of symmetry and has an octahedral geometry that is distorted both by the presence of four-membered chelate rings and by the Jahn-Teller effect. The equatorial plane is formed by the N atoms of two aminopyrimidine (ampym) ligands and by the weakly basic carbonyl O atoms of the acesulfamate ligands, while the more basic deprotonated N atoms of these ligands are in the elongated axial positions with a strong misdirected valence. The crystal is stabilized by pyrimidine ring stacking and by intermolecular hydrogen bonding involving the NH2 moiety of the ampym ligand and the carbonyl O atom of the acesulfamate moiety. PMID- 15876706 TI - Two square-pyramidal chromium(V)-nitride complexes: bis(2-methylquinolin-8 olato)nitridochromium(V) and nitridobis(2-sulfidopyridine N-oxide)chromium(V). AB - Two new chromium(V)-nitride complexes with a coordination sphere completed by bidentate ligands have been synthesized and structurally characterized. Bis(2 methylquinolin-8-olato)nitridochromium(V), [Cr(C10H8NO)2(N)], has the coordination sphere completed by an equatorial N2O2 set of ligators. The compound crystallizes with the five-coordinate complexes at sites with twofold rotational symmetry and all Cr-N bond directions aligned with the crystallographic b axis. Nitridobis(2-sulfidopyridine N-oxide)chromium(V), [Cr(C5H4NOS)2(N)], crystallizes with the molecules on general positions and has an equatorial S2O2 coordination environment, which is unprecedented among nitride complexes of the first-row transition metals. In both systems, Cr[triple-bond]N bonds are short at ca 1.56 A. PMID- 15876707 TI - Single-crystal neutron-diffraction study of 3.4% Zn-doped (ND4)2[Cu(D2O)6](SO4)2 at 20 K. AB - Doping the perdeuterated ammonium copper Tutton salt (ND4)2[Cu(D2O)6](SO4)2 [perdeuterated diammonium hexaaquacopper(II) bis(sulfate)] with Zn leads to a change in the structure from dimorph A (low density) to dimorph B (high density). This change, which accompanies a switch in the direction of the Jahn-Teller distortion, had previously been observed to occur with substitution of Zn2+ at the Cu2+ site of between 1.3 (A) and 3.4% (B). In this study, the single-crystal neutron-diffraction analysis of (ND4)2[(Cu/Zn)(D2O)6](SO4)2 at 20 K, with 3.4% Zn doping and a deuterium substitution of 85% on the H-atom sites, reveals that the structure is entirely of type B, with the Cu/Zn site at an inversion centre and with no evidence of disorder or unusual atomic displacement parameters that might occur near a phase transition boundary. PMID- 15876708 TI - [catena-Poly[[[(N,N,N',N'-tetramethylethylenediamine)copper(II)]-mu-oxalato] tetrahydrate]]. AB - In the title compound, {[Cu(C2O4)(C6H16N2)].4H2O}n, the Cu atom is six-coordinate in a distorted octahedral geometry, and N,N,N',N'-tetramethylethylenediamine acts as a chelating ligand. The oxalate anion, which lies about an inversion centre, bridges the metal cations and forms infinite chains. The structure cohesion is ensured by hydrogen-bonding interactions, which form a three-dimensional framework. PMID- 15876709 TI - Two Cd and Hg peroxodisulfate complexes with aromatic amines. AB - The crystal structures of two complexes containing the peroxodisulfate anion are reported, namely mu-peroxodisulfato-1kappaO:2kappaO'-bis[(acetato kappa2O,O')aqua(2,2':6',2''-terpyridine-kappa3N,N',N'')cadmium(II)] heptahydrate, [Cd2(C2H3O2)2(S2O8)(C15H11N2)2(H2O)2].7H2O, (I), and catena-poly[[[bis(2,2' bipyridine-kappa2N,N')mercury(II)]-mu-peroxodisulfato-kappa2O:O'] 0.4-hydrate], {[Hg(C10H8N2)2(S2O8)].0.4H2O}n, (II). In both structures, the anion behaves as a bridge, linking neighbouring coordination polyhedra in two different ways, either tightly bound to the heptacoordinated Cd2+ cation forming neatly separated dimeric entities in (I) or across a shorter O-S-O path producing weakly connected chains by way of 'semicoordination' to the Hg2+ cations in (II). PMID- 15876710 TI - Cyanocobalt(III) complexes of penta- and tetradentate-coordinated macrocyclic hexaamines. AB - The pendent-arm macrocyclic hexaamine trans-6,13-dimethyl-1,4,8,11 tetraazacyclotetradecane-6,13-diamine (L) may coordinate in tetra-, penta- or hexadentate modes, depending on the metal ion and the synthetic procedure. We report here the crystal structures of two pseudo-octahedral cobalt(III) complexes of L, namely sodium trans-cyano(trans-6,13-dimethyl-1,4,8,11 tetraazacyclotetradecane-6,13-diamine)cobalt(III) triperchlorate, Na[Co(CN)(C13H30N6)](ClO4)3 or Na{trans-[CoL(CN)]}(ClO4)3, (I), where L is coordinated as a pentadentate ligand, and trans-dicyano(trans-6,13-dimethyl 1,4,8,11-tetraazacyclotetradecane-6,13-diamine)cobalt(III) trans-dicyano(trans 6,13-dimethyl-1,4,8,11-tetraazacyclotetradecane-6,13-diaminium)cobalt(III) tetraperchlorate tetrahydrate, [Co(CN)2(C14H32N6)][Co(CN)2(C14H30N6)](ClO4)4.4H2O or trans-[CoL(CN)2]trans-[Co(H2L)(CN)2](ClO4)4.4H2O, (II), where the ligand binds in a tetradentate mode, with the remaining coordination sites being filled by C bound cyano ligands. In (I), the secondary amine Co-N bond lengths lie within the range 1.944 (3)-1.969 (3) A, while the trans influence of the cyano ligand lengthens the Co-N bond length of the coordinated primary amine [Co-N = 1.986 (3) A]. The Co-CN bond length is 1.899 (3) A. The complex cations in (II) are each located on centres of symmetry. The Co-N bond lengths in both cations are somewhat longer than in (I) and span a narrow range [1.972 (3)-1.982 (3) A]. The two independent Co-CN bond lengths are similar [1.918 (4) and 1.926 (4) A] but significantly longer than in the structure of (I), again a consequence of the trans influence of each cyano ligand. PMID- 15876711 TI - catena-Poly[copper(II)-mu-L-tyrosyl-L-leucinato]. AB - In the title compound, [Cu(C15H20N2O4)]n, the copper(II) coordination is square planar. The anionic L-tyrosyl-L-leucinate ligand binds in an N,N',O-tridentate mode to one Cu(II) cation on one side and in an O-monodentate mode to a second Cu(II) cation on the other side, thus defining -Cu-O-C-O-Cu'- chains which run along the a axis. These chains are held together by a strong hydrogen bond involving the hydroxy H atom. PMID- 15876712 TI - Imino Diels-Alder adducts. II. Two pyrano[3,2-c]quinolines. AB - 8-Chloro-9-fluoro-5-phenyl-3,4,4a,5,6,10b-hexahydro-2H-pyrano[3,2-c]quinoline and 10-chloro-9-fluoro-5-phenyl-3,4,4a,5,6,10b-hexahydro-2H-pyrano[3,2-c]quinoline, both C18H17ClFNO, are diastereoisomers, formed as the result of the imino Diels Alder reactions of N-benzylideneanilines with 3,4-dihydro-2H-pyran. The crystal structures reveal the stereochemistry of the pyran ring, which is endo/exo to the quinoline ring system formed in the cycloaddition step. In both structures, the pyran ring adopts a chair conformation, while the nitrogen-containing heterocyclic ring prefers a half-chair conformation. The structures differ essentially in the relative orientation of the ring junction H atoms. PMID- 15876713 TI - 2-(2-Acetylamino-5-chlorophenyl)-2,2-difluoroethanoic acid and 2-(2-acetylamino-5 methylphenyl)-2,2-difluoroethanoic acid, and 2-(2-acetylaminophenyl)-2,2-difluoro N-phenylacetamide and 2-(2-acetylaminophenyl)-N-(4-chlorophenyl)-2,2 difluoroacetamide: examples of variation in molecular packing and hydrogen bonding motif induced by substituent change. AB - Among the title compounds, viz. the acids C10H8ClF2NO3, (I), and C11H11F2NO3, (II), and the amides C14H14F2N2O2, (III), and C14H13ClF2N2O2, (IV), the change of substituent from Cl in (I) to methyl in (II) has a dramatic effect upon the hydrogen bonding between the molecules, which occur in layers in both cases. In the structures of (III) and (IV), hydrogen bonds connect the molecules to form chains, but the introduction of a chloro substituent in (IV) has a profound effect on the orientation of the molecules within the chains and the packing of the chains in the structure as a whole. PMID- 15876714 TI - Supramolecular structures of four isomorphous anilinium 2-carboxy-4-nitrobenzoate salts: 4-X-C6H4NH3+.C8H4NO6- (X = H, Cl, Br and I). AB - Anilinium 2-carboxy-4-nitrobenzoate, C6H8N+.C8H4NO6-, (I), 4-chloroanilinium 2 carboxy-4-nitrobenzoate, C6H7ClN+.C8H4NO6-, (II), 4-bromoanilinium 2-carboxy-4 nitrobenzoate, C6H7BrN+.C8H4NO6-, (III), and 4-iodoanilinium 2-carboxy-4 nitrobenzoate, C6H7IN+.C8H4NO6-, (IV), are approximately isostructural. In each compound, the ions are linked into complex sheets by a combination of O-H...O and N-H...O hydrogen bonds. Within the sheets, two distinct one-dimensional substructures can be identified, viz. a chain of edge-fused R(3)3(13) rings and a double helix of simple C(2)2(9) chains. In (I) and (IV), the sheets are linked by a C-H...O(nitro) hydrogen bond and a two-centre C-I...O(nitro) interaction, respectively, but the corresponding C-Cl...O and C-Br...O contact distances in (II) and (III) are not significantly shorter than the sum of the van der Waals radii. PMID- 15876715 TI - Ethyl 5-amino-3-(4,6-dimethylpyrimidin-2-ylamino)-1-methyl-1H-pyrazole-4 carboxylate and ethyl 5-amino-3-(4,6-dimethylpyrimidin-2-ylamino)-1-(2 nitrophenylsulfonyl)-1H-pyrazole-4-carboxylate. AB - The molecular structures of ethyl 5-amino-3-(4,6-dimethylpyrimidin-2-ylamino)-1 methyl-1H-pyrazole-4-carboxylate, C13H18N6O2, (I), and ethyl 5-amino-3-(4,6 dimethylpyrimidin-2-ylamino)-1-(2-nitrophenylsulfonyl)-1H-pyrazole-4-carboxylate, C18H19N7O6S, (II), have been determined. There are two intramolecular N-H...O bonds and one intermolecular N-H...O hydrogen bond in (I). The rings formed by the N-H...O hydrogen bonds are almost planar. In (II), three intramolecular N H...O hydrogen bonds exist. PMID- 15876716 TI - (E,E)-2,5-Dipropoxy-1,4-bis[2-(2,4,6-trimethoxyphenyl)ethenyl]benzene. AB - The title compound, C30H34O8, crystallizes in the space group P-1 with one-half of a molecule in the asymmetric unit. A three-dimensional network is generated by OCH3...pi and CH...pi interactions. The conformation of the C-C bond exocyclic to the central benzene ring is different from that of every other known derivative. A comparison of the geometry of the title molecule and of its solid-state structure with other 2,4,6-trimethoxy-substituted PPV [i.e. poly(p phenylenevinylene)] oligomers, in particular the isopropoxy-substituted compound, is provided. PMID- 15876717 TI - Polymorphism of (Z)-4-bromo-N-(pent-2-enyl)-N-(3-phenylprop-2 ynyl)benzenesulfonamide. AB - The title compound, C20H20BrNO2S, has two polymorphic crystal structures with very similar lattice constants. A number of crystals are composites of the two polymorphs. Both crystal structures contain identical layers of molecules. The packing of the layers, however, is different for the two polymorphs. PMID- 15876718 TI - A main-group donor-acceptor adduct: 1,3,4,5-tetramethylimidazol-2-ylidene tris(pentafluorophenyl)borane. AB - The solid-state structural analysis of the title compound, Me4C2N2C-->B(C6F5)3 or C25H12BF15N2, provides useful metric parameters for the qualitative comparison of the donor strength with other mono- and dicoordinate main-group compounds which feature distinctive lone-pair character. PMID- 15876719 TI - 2-[Bis(pyrazol-1-yl)methyl]-4-tert-butyl-6-(phenylsulfanyl)phenol. AB - The title compound, C23H24N4OS, contains a highly asymmetric bifurcated intramolecular hydrogen bond between the hydroxy group and two pyrazole N atoms. The compound associates into centrosymmetric dimers in the crystal through two unique C-H...pi interactions, which are in turn linked into a (6,3)-network through an additional intermolecular C-H...N hydrogen bond. PMID- 15876720 TI - 5-Butyl-5-ethylbarbituric acid: a phase transition at low temperature. AB - The room-temperature crystal structure of 5-butyl-5-ethylbarbituric acid (generally known as butobarbitone), C10H16N2O3, was reported in space group C2/c [Bideau (1971). C. R. Acad. Sci. Paris Ser. C, 272, 757-760]. A redetermination at 120 K using synchrotron radiation shows the space group at this temperature to be P2(1)/n and not C2/c. There are two crystallographically independent molecules in the asymmetric unit, but no solvent. Reported issues concerning possible disorder of the molecule are addressed; the butyl substituent of one of the molecules adopts an unusual conformation in being not fully extended. A subsequent re-collection at room temperature shows that the space group is indeed C2/c (A2/a with the axes selected in this report), and so the crystal structure undergoes a phase change upon cooling to 120 K. PMID- 15876721 TI - Pseudomerohedrally twinned crystal structure of 2,3-diphenylbuta-1,3-diene. AB - The title compound, C16H14, is twinned by reticular pseudomerohedry of twin index 2. The primitive monoclinic cell of the single crystal can be transformed into a B-centred pseudo-orthorhombic supercell with a fourfold volume. The twofold twin operation about the reciprocal a* axis of the primitive monoclinic cell is co directional with the approximate C2 axis of the molecule and a symmetry element of the orthorhombic supercell. A tentative twin domain model is proposed. PMID- 15876722 TI - 1,3-Bis(4-methylphenyl)triazene, 1-(4-chlorophenyl)-3-(4-fluorophenyl)triazene and 1-(4-fluorophenyl)-3-(4-methylphenyl)triazene. AB - The title 4,4'-disubstituted diphenyl-1,3-triazines, C14H15N3, (I), C12H9ClFN3, (II), and C13H12FN3, (III), each contain a triazene group (-N=N-NH-) having an extended conformation. The dihedral angles between the two benzene rings in (I), (II) and (III) are 4.3, 3.4 and 6.5 degrees , respectively. The molecules are almost entirely planar, with maximum deviations from the mean planes of 0.1087 (2), -0.1072 (7) and 0.1401 (3) A, respectively. In each compound, the molecules are linked by N-H...N hydrogen bonds to form chains and pack similarly in the crystal structures. PMID- 15876723 TI - Supramolecular structures of five 5-(arylmethylene)-1,3-dimethylpyrimidine 2,4,6(1H,3H,5H)-triones: isolated molecules, hydrogen-bonded chains and chains of fused hydrogen-bonded rings. AB - In each of the five title compounds, namely 5-benzylidene-1,3-dimethylpyrimidine 2,4,6(1H,3H,5H)-trione, C13H12N2O3, (I), 5-(3-methoxybenzylidene)-1,3 dimethylpyrimidine-2,4,6(1H,3H,5H)-trione, C14H14N2O4, (II), 5-(4 methoxybenzylidene)-1,3-dimethylpyrimidine-2,4,6(1H,3H,5H)-trione, C14H14N2O4, (III), 5-[4-(dimethylamino)benzylidene]-1,3-dimethylpyrimidine-2,4,6(1H,3H,5H) trione, C15H17N3O3, (IV), and 5-(3,5-di-tert-butyl-4-hydroxybenzylidene)-1,3 dimethylpyrimidine-2,4,6(1H,3H,5H)-trione, C21H28N2O4, (V), which crystallizes with Z' = 2 in P-1, there is a very wide C-C-C angle at the methine C atom linking the two rings, ranging from 137.1 (2) degrees in (I) to 139.14 (14) degrees in (III). There is evidence for intramolecular charge separation in (IV) and, to a lesser degree, in (III). The molecules of (I)-(III) are linked by pairs of C-H...O hydrogen bonds into chains of edge-fused rings, with alternating R(2)2(14) and R(2)2(16) rings in (I), alternating R(2)2(14) and R(4)4(20) rings in (II), with two types of R(2)2(16) rings alternating in (III). The molecules in (IV) are linked by a single C-H...O hydrogen bond into simple C(8) chains, but there are no direction-specific intermolecular interactions in (V). PMID- 15876724 TI - Supramolecular structures of three isomeric (E,E)-1-(2-iodophenyl)-4 (nitrophenyl)-2,3-diaza-1,3-butadienes: changes in intermolecular interactions consequent upon changes of substituent location. AB - The supramolecular structures of the three isomeric (E,E)-1-(2-iodophenyl)-4 (2/3/4-nitrophenyl)-2,3-diaza-1,3-butadienes, C14H10IN3O2, are compared. In the 2 nitro isomer, the molecules are disordered across centres of inversion in space group C2/c and are linked into chains by a two-centre iodo-nitro interaction. The molecules of the 3-nitro isomer are linked into a three-dimensional framework by a combination of C-H...O and C-H...I hydrogen bonds and aromatic pi-pi stacking interactions, while molecules of the 4-nitro isomer are linked into sheets by a C H...O hydrogen bond and a two-centre iodo-nitro interaction. PMID- 15876725 TI - The alpha2-polymorph of salicylideneaniline. AB - N-Salicylideneaniline (SA), C13H11NO, belongs to the large family of aromatic Schiff bases. It is of particular importance owing to its reversible photoreactivity. SA forms two photochromic polymorphs, both with two non-coplanar benzene rings. In addition, we have recently discovered a planar polymorph, named the beta-polymorph, which will be discussed in a subsequent paper. We report here the structure of the alpha2-polymorph in the orthorhombic crystal system. This compound exhibits a strong intramolecular O-H...N hydrogen bond and the dihedral angle between the two rings varies with temperature. PMID- 15876726 TI - (2-Hydroxyethyl)hydrazinium(2+) dichloride. AB - The crystal structure of the title compound, C2H10N2O(2+).2Cl-, is built up from one 2-hydroxyethylhydrazinium(2+) cation and two Cl- anions. The molecular structure is stabilized by O-H...Cl and N-H...Cl hydrogen bonds. The crystal structure is stabilized by one N-H...O and three N-H...Cl interactions, and the three-dimensional network of hydrogen bonds stabilizes the crystal packing. All five hydrazinium H atoms are involved in hydrogen bonds to Cl- anions. The Cl...H contact distances range from 2.122 (15) to 2.809 (14) A. PMID- 15876727 TI - 4-[Bis(4-hydroxy-3,5-dimethylphenyl)methyl]pyridinium chloride and bromide. AB - The isostructural title salts, 4-[bis(4-hydroxy-3,5 dimethylphenyl)methyl]pyridinium chloride and bromide, C22H24NO2+.Cl- and C22H24NO2+.Br-, exhibit extended hydrogen-bonded ribbons in the solid state. The halide ions form hydrogen bonds to the pyridinium NH+ group and to the phenol OH groups of the cation. These bonds are coplanar to within 0.1 A and form a T configuration at the anion. PMID- 15876728 TI - A new polymorph and two inclusion compounds of 9,9'-spirobifluorene. AB - Two new inclusion compounds of 9,9'-spirobifluorene (SBF) incorporating benzene [(I), C25H16.C6H6] and biphenyl [(II), C25H16.C12H10], and a new polymorph of SBF [(III), C25H16] are reported. All three exhibit C-H...pi(arene) hydrogen bonds between adjacent SBF molecules. Compound (II) also contains biphenyl C-H to SBF pi-arene interactions. Collectively, hydrogen bonding gives rise to a chain and a layered motif in compounds (I) and (II), respectively. PMID- 15876729 TI - 3,3a-Dihydrocyclopenta[b]chromen-1(2H)-ones from the reaction of salicylaldehyde and 2-cyclopenten-1-one. AB - The two title chromene compounds, 3,3a-dihydrocyclopenta[b]chromen-1(2H)-one, C16H12O2, (I), and 2-(2-hydroxybenzylidene)-3,3a-dihydrocyclopenta[b]chromen 1(2H)-one, C19H14O3, (II), have been determined in the monoclinic space group P2(1)/n. Compound (I) is mainly stabilized by C-H...pi interactions. Compound (II) is linked into infinite one-dimensional chains with a C(3) motif via intermolecular O-H...O hydrogen bonds. The intermolecular C-H...pi and pi-pi interactions also play key roles in stabilizing the crystal packing. Two intramolecular C-H...O hydrogen bonds with S(5) motifs were detected in (II). PMID- 15876730 TI - Hydrogen bonding in C-substituted nitroanilines: simple C(8) chains in 2-bromo-6 chloro-4-nitroaniline. AB - In the title compound, C6H4BrClN2O2, the Br and Cl substituents are disordered over two sites, and the intramolecular dimensions provide evidence for charge polarization. The molecules are linked into C(8) chains by a single N-H...O hydrogen bond. PMID- 15876731 TI - Supramolecular aggregation of two hydroxycarboxylic acid derivatives. AB - The crystal structures of 7,7-dicyclobutyl-5-hydroxymethyl-6 oxabicyclo[3.2.1]octane-1-carboxylic acid, C17H26O4, (I), and 1-(hydroxymethyl)-7 oxaspiro[bicyclo[3.2.1]octane-6,1'-cyclopentane]-5-carboxylic acid, C13H20O4, (II), determined at 170 K, show that the conformation of the hydroxymethyl group (anti or gauche) affects the dimensionality (one- or two-dimensional) of the supramolecular structures via O-H...O hydrogen bonds. In (I), the carboxyl and hydroxymethyl groups interact with themselves, forming a one-dimensional step ladder, while in (II), a two-dimensional structure is made up of carboxylic acid centrosymmetric R(2)2(8) dimers connected by hydroxyl-to-ether contacts. PMID- 15876732 TI - N1-(4-Bromophenyl)-N2-hydroxy-2-oxo-2-phenylacetamidine. AB - In the title compound, C14H11BrN2O2, which has the oxime group in an E conformation, molecules are linked by strong O-H...O and N-H...O hydrogen bonds into chains of edge-fused rings, unlike closely related compounds. PMID- 15876733 TI - [Change in the definition of osteoporosis especially on bone quality]. AB - In recent years the term Bone Quality has been used with increasing frequency in the field of osteoporosis diagnosis and treatment. Bone quality is an umbrella term which describes a set of characteristics that influence bone strength. The strength of bone depends on its structure and material properties which are affected by bone remodeling and modeling. The structural property is determined by the size and shape of bone and also the microarchitecture. Material property can be determined by mineral crystallinity, collagen structure and micro-damage in bone. Osteoporosis is defined as a skeletal disorder characterized by compromised bone strength predisposing to an increased risk of fracture. Bone strength reflects the integration of two main features: bone density and bone quality. PMID- 15876734 TI - [Molecules which are involved in osteoclastic bone resorption: from the aspect of targets of treatment for osteoporosis]. AB - Bone in vertebrates is continuously destroyed and reformed to maintain bone volume throughout their life. In normal bone remodeling, osteoblastic bone formation follows osteoclastic bone resorption in a precise and quantitative manner. In osteoporosis, the bone resorption surpasses the bone formation. Recent studies have shown that osteoclastic bone resorption is tightly regulated by bone forming osteoblasts. Osteoblasts express RANKL (receptor activator of nuclear factor-KappaB ligand), which plays central roles in osteoclast differentiation and function. In this review article, we explain molecules which are involved in the RANKL-RANK interaction in the regulation of osteoclastic bone resorption. PMID- 15876735 TI - [Genes involved in bone formation as a therapeutic target for osteoporosis]. AB - Osteoblasts are derived from mesenchymal stem cells, common progenitors for adipocytes and other lineages. While transcription factors such as Runx2/Cbfa1 and Osterix are absolutely required for osteoblast differentiation, simply increasing their expression does not necessarily lead to increased bone formation. In contrast, genes such as deltafosB and an AP-1 target, interleukin 11, which have been shown to enhance bone formation in vivo and to be induced by mechanical stress and PTH, can be better therapeutic targets to develop drugs that stimulate bone formation. PMID- 15876736 TI - [Searches for the genes determining bone mineral density with gene polymorphisms]. AB - The genes determining bone mineral density have been searched by association and linkage studies using gene polymorphisms. Candidate gene approaches as well as genome-wide studies with single nucleotide polymorphisms have produced many products. In addition, rare mutations causing monogenic hereditary bone diseases gave novel aspects for the genes contributing the variation in bone mineral density. Furthermore, studies of functional SNPs would explain the variation in bone mineral density and basic pathophysiology of osteoporosis. PMID- 15876737 TI - [Genetic analysis of osteoporosis susceptibility: future perspectives on genome wide analysis]. AB - Genetic susceptibility of osteoporosis has been analyzed mainly via sib-pair analysis and association study on candidate genes, however these approaches might have substantial limitation. Recently proposed genome wide analysis of association using high density polymorphic markers has not been established on this field. Preferable phenotypes and suitable subjects have to be defined as well as for tag-SNP selection and the method of multi-step screening. PMID- 15876738 TI - [Methods of statistical genetics and use of database for genome information]. AB - Knowledge and technology of bioinformatics have become inevitable for gene and genome research. Education and research in this field of science are not sufficient in Japan. There are two different approaches to trait mapping, the way by which traits are mapped on the genome. Thus, the knowledge-based approach uses functions of molecules while the statistics-based approach uses polymorphisms. Statistics-based approach uses two different methods, linkage analysis and analysis based on linkage disequilibrium. Various phenotypes are efficiently mapped on the genome using such methods. Recently, bioinformatic data base search is mostly performed using internet. Anyone can perform sequence-search, homology search and SNP-search. Since such data bases change quickly, readers should access the databases themselves and be used to the procedures for them. PMID- 15876739 TI - [Systemic bone diseases; clues for the pathogenetic mechanism of osteoporosis]. AB - Single gene diseases may give us lots of clues to clarify the physiological roles of specific genes in human body. In the case of bone, extensive analysis of the pathogenetic mechanisms of systemic bone diseases may provide new important information on the factors in bone metabolism. In this manuscript, pathogenetic mechanisms of osteoporosis-pseudoglioma syndrome, osteogenesis imperfecta, homocystinuria, lysinuric protein intolerance and Menkes disease will be reviewed in relation to the pathogenesis of osteoporosis. PMID- 15876740 TI - [Novel phosphate regulating genes and osteoporosis]. AB - The hormones currently believe to influence inorganic phosphate (Pi) metabolism are parathyroid hormone (PTH) and the active metabolite to vitamin D. A new class of phosphate-regulating factors, collectively known as the phosphatonins have been shown to be associated with the hypophosphatemic diseases. The reabsorption of Pi in the kidney is a major determinant of the plasma Pi level. Reabsorption is largely regulated by the type II a sodium-dependent Pi cotransporter (NPT2a) that is expressed in renal proximal tubular cells. Phosphatonins cause Pi wasting by controlling the amount of NPT2a on the apical surface of the proximal tubular cell. A recent finding indicates that mutations in NPT2a can contribute to nephrolithiasis and osteoporosis in humans and suggests that changes in NPT2a levels may also cause other human disease. We discuss the roles of phosphatonins and NPT2a in bone formation. PMID- 15876741 TI - [Bone metabolism in metabolic syndrome and its treatment]. AB - Metabolic syndrome has recently been identified to be involved in the development of multiple diseases, such as type 2 diabetes (or glucose intolerance), dyslipidemia and hypertension. Its principal features are obesity and insulin resistance. Bone metabolism is a complex process that is regulated by complicated local and systemic factors. Recent investigations demonstrate an involvement of sympathetic nervous system and glucose and lipid metabolism in bone metabolism. Obesity and insulin resistance may favor bone dynamics, whereas diabetes and dyslipidemia may not. More importantly, various drugs for these disorders have been reported to have beneficial effects on bone health. On clinical situation to manage metabolic syndrome, physicians may want to evaluate outcome of their practice for bone metabolism. PMID- 15876742 TI - [Central control of bone metabolism]. AB - Leptin, a well-known adipokine regulating energy metabolism, inhibits bone formation via sympathetic nervous system. Recently, it was reported that adrenergic beta(2) receptor-deficient mice have high bone mass due to increased bone formation and decreased bone resorption. In addition, cocaine- and amphetamine-regulated transcript, an anorexigenic neuropeptide whose expression is regulated by leptin, has been shown to inhibit bone resorption. Thus, leptin regulates not only bone formation but bone resorption centrally. PMID- 15876743 TI - [Improvement of musculoskeletal function by cell-based therapy using mesenchymal stem cells with a prolonged life span]. AB - Cell transplantation has recently been attempted to improve musculoskeletal function. Many types of cells, such as embryonic stem cells, fetal cardiomyocytes, myoblasts, bone marrow hematopoietic cells, and mesenchymal stem cells (MSCs), have been transplanted to functionally restore damaged or diseased tissue in animal models, and marrow-derived mononuclear cells have been injected into ischemic limb clinically. MSCs can be a useful source of cell transplantation for several reasons:they have the ability to proliferate and differentiate into mesodermal tissues, including myocytes, they entail no ethical or immunological problems, and bone marrow aspiration is an established routine procedure. When placed in appropriate in vitro and in vivo environments, MSCs can give rise to all major mesenchymal tissues, such as bone, cartilage, muscle, and adipose tissue. Direct injection of murine and porcine MSCs into skeletal muscles has been shown to be feasible in murine models of ischemic limb. Large numbers of cells must be injected into damaged sites in ischemic limb to restore muscular function in humans, and cells need to be injected into the entire limb. Until now, however, there have been no reports of a sufficient number of differentiated human myocytes ever having been obtained to restore muscular function of ischemic limb. One of the reasons for this is that the life span of human cells in vitro is limited. Human cells reach senescence or stop cell growth after a limited number of cell replications, and the average number of hMSC population doublings (PDs) has been found to be 38, implying that it would be difficult to obtain enough cells to restore the function of ischemic limb. To resolve these problems and to establish a model of cell-based therapy, prolongation of the life span of hMSCs without affecting differentiation capability is essential. PMID- 15876744 TI - [Wnt signaling and bone metabolism]. AB - The Wnt family of secreted signaling molecules is essential in embryo development and tumor formation. In addition to the Frizzled family of serpentine receptors, LRP5, a member of the LDL receptor family, was recently shown to function as Wnt receptor. The finding of unique mutations in the human LRP5 gene as a modulator of bone mass establishes the importance of this novel and unexpected role of Wnt/LRP5 signaling in bone development and bone metabolism. PMID- 15876745 TI - [Bisphosphonates: the molecular targets and mechanisms of action]. AB - Bisphosphonates directly act on osteoclasts to inhibit bone resorption and used most widely to treat osteoporosis. The compounds can be classified into two groups with different modes of action. Nitrogen containing bisphosphonates exert their effects by inhibiting a key enzyme in the mevalonate pathway. The specific target is the isoprenoid biosynthetic enzyme, farnesyl pyrophosphate synthase, which is indispensable for protein prenylation and activation of intracellular signalling proteins, including the small GTPases Rho, Rac, Cdc42 and Ras. The disruption of the function of these key enzymes may explain the loss of osteoclast activity and induction of apoptosis. Whereas the first generation of bisphosphonates such as etidronate and clodronate (nitrogen deficient bisphosphonates) can be incorporated into nonhydrolysable analogues of ATP that may inhibit ATP-dependent intracellular events. Bisphosphonates are highly effective to inhibit bone resorption and increase bone mineral density, although their precise mechanisms of molecular action are not completely understood. PMID- 15876746 TI - [Molecular mechanism of tissue-specific actions of sex steroid hormones and SERM for bone remodeling]. AB - Female sex steroid hormone, estrogen, exerts a wide variety of biological actions including lipid metabolism and bone remodeling. Moreover, estrogen is known to be involved in many pathological states like osteoporosis, tumor incidence and tumor development of female reproductive organs. Most of such estrogen actions are believed to mediate transcriptional controls of a particular set of target genes by its nuclear receptor, estrogen receptor (ER). The products of the genes regulated by ER are responsible for the overt estrogen actions in the many estrogen target tissues. Reflecting the estrogen actions, two ER subtypes (alpha, beta) express in the bone and the other target tissues. ER is a hormone-dependent transcriptional factor and hormone binding is known to alter the structure of ER protein, following dissociation/recruitment of multiple co-regulator complexes. Synthetic ER ligands, SERM, are believed to cause the ligand type-specific alteration in ER structure. This SERM-induced structural alteration acquires the tissue-specific function of liganded ER, as SERM acts as an agonist for bone remodeling. PMID- 15876747 TI - [The genes related to the effects of vitamin D on muscle and bone]. AB - Vitamin D is most frequently used as a drug for osteoporosis in Japan, and its effects on the maintenance of bone mineral density and inhibition of fracture risk have been shown. Recently, the actions of vitamin D on muscle have been elaborated, and its prevention of falls is noted. Active vitamin D modulates several genes in muscle and bones. The studies using vitamin D receptor deletion mice revealed that active vitamin D modulates the genes related to muscle differentiation and bone resorption. PMID- 15876748 TI - [Molecular mechanisms of vitamin K action in the bone homeostasis]. AB - Vitamin K is used as an anti-osteoporosis drug in Japan. Moreover, vitamin K intake has been found to decrease hip fracture risk. In the bone homeostasis, vitamin K action is mediated through two molecular mechanisms: posttranslational modification of proteins, and regulation of gene expression. The former is vitamin K-dependent carboxylation, in which vitamin K functions as an essential cofactor for modification of glutamic acid residues to gamma-carboxyglutamic acid residues. The latter is a novel mechanism that regulates the transcription of target genes by vitamin K through activation of steroid and xenobiotic receptor (SXR). The two mechanisms may coordinately contribute to vitamin K function in the bone metabolism. PMID- 15876749 TI - [Parathyroid hormone -- its mechanisms of action and issues on clinical application]. AB - PTH, recently approved for severe osteoporosis in US and Europe, acts as an anabolic agent for bone when given intermittently. However, when continuously given, PTH acts as a catabolic agent for bone, as seen in primary hyperparathyroidism. The mechanisms by which PTH acts in opposite directions are largely unknown. In this review, some of the possible mechanisms will be discussed. PMID- 15876750 TI - [Osteoporosis and intake of vitamins]. AB - Subclinical vitamins deficiency is common in the elderly, especially in osteoporotic patients. However, most physicians in this area are just focused on drugs for the treatment of osteoporosis. It is already established that several vitamins influence bone turnover, bone mineral density, or even the risk of hip fractures. Improving these vitamins status may help to treat and prevent osteoporosis in elderly people. Recently higher vitamin D intake is recognized to be needed to keep not only bone health but also muscle strength. More sun exposure might be needed for improved bone health in the elderly. Deficiency of Vitamin K, C, or B(12) may be also important modifiable risk factors for osteoporosis and bone fracture. Excessive retinal supplementation may become associated with higher bone loss. Thus such diet rich in fruit and vegetables together with fish and meat could fulfill a balance among these vitamins and should be recommended for prevention or treatment of osteoporosis. PMID- 15876751 TI - [Jaws of birds]. AB - Birds (Class Aves) have been hypothesized to have evolved from theropod dinosaurs, which belong to the group known as diapsid reptiles, during the Mesozoic era. They subsequently acquired derived characters relating to flying ability in all parts of their body, including the jaws, which were toothed in the ancestral forms but beaked in later members. When the fore limbs became modified as wings specialized for flight, they became incapable of grasping food, which function was then assumed by the avian bill. Avian bills consequently evolved great morphological diversity reflecting very divergent modes of feeding. To illustrate the progression of "the evolution of vertebrate jaws," the comparative osteology of avian cranial bones is exemplified by reference to Mesozoic birds and the morphological and functional variations observed among the bills of Neornithes. PMID- 15876752 TI - [Two cases of hemodialysis patients with absolute hypoparathyroidism or normal parathyroid function who were suffering from prostate carcinoma]. AB - In recent years, the incidence of prostate carcinoma in patients with renal failure is increasing. But it is difficult to diagnose the occurrence of this neoplasm because of decrease of urinary volume in these patients. Two hemodialysis patients (age: 64, 68 years, the duration of hemodialysis: 10, 8 years) suffering from prostate carcinoma were treated with androgen deprivation therapy, which accelerates bone resorption. (This kind of osteoporosis is often treated by bisphosphonate, but this treatment is prohibited in hemodialysis patients.) Both serum testosterone and estradiol went down to the extremely lower levels in these patients. Thereafter, one (case 2) of them was maintained under absolute hypoparathyroidism by the conservative therapy with vitamin D. As a result, bone mineral density of the patient (case 2) was well maintained during one year. In conclusion, hypoparathyroidism was helpful to prevent bone loss in patients receiving androgen deprivation therapy. PMID- 15876753 TI - Reproductive health education intervention trial. AB - OBJECTIVE: To measure the effectiveness of a reproductive health education package in improving the knowledge of adolescent girls aged 15-19 years in Chandigarh (India). METHODS: A reproductive health education package, developed in consultation with parents, teachers and adolescents, was delivered to randomly sampled classes of two senior secondary schools and one school was selected as control. In one school, a nurse conducted 15 sessions for 94 students in three batches using conventional education approach. In another school she conducted sessions for a selected group of 20 adolescents who later disseminated the messages informally to their 84 classmates (peer education). Using a 70-item structured questionnaire the knowledge of 95 adolescents from conventional, 84 from peer, and 94 from control school were assessed before and one month after the last session. Change in the score in intervention and control groups was tested by ANOVA taking age and socio-economic status as covariates. RESULTS: Teachers, parents and students overwhelmingly (88%, 95.5% and 93% respectively) favoured reproductive health education program. Five percent of the respondents reported that someone in their class is having sexual relations, and 13% of the girls approved of pre-marital sexual relations. Reproductive health knowledge scores improved significantly after intervention in conventional education (27.28) and peer education group (20.77) in comparison to the controls (3.64). Post-test scores were not significantly different between peer education group and conventional education group (43.65 and 40.52 respectively) though the time consumed in delivering the peer education intervention was almost one third of the time taken to implement conventional education. CONCLUSION: Peer education and conventional education strategies were effective in improving the reproductive health knowledge of adolescent girls but peer strategy was less time consuming. PMID- 15876754 TI - Long-term outcome in coma. AB - OBJECTIVE: To assess the relationship between Modified Glasgow Coma Scale, its components, brain stem reflexes and long term functional outcome in children with acute non-traumatic coma. METHOD: MGCS and brainstem reflexes were assessed at 6 hourly intervals for 72 hours from the time of admission. The children were followed up regularly and functional outcome was assessed at the end of 9 months. The lowest score of the MGCS and worst brain stem reflexes were used for the analysis. RESULTS: Higher total MGCS score and verbal response score had a significant positive correlation with better functional outcome as measured by GOS and intelligence quotient. There was no association between the language function and the initial MGCS, its components and brain stem reflexes. Lower verbal response (P = 0.005) was the only factor that was found to be individually associated with poorer long term GOS score and intelligence quotient by multivariate linear regression analysis. CONCLUSION: In the long term prediction of outcome in acute non-traumatic coma, MGCS is not useful. However, verbal response, a component of MGCS, correlates well with long term functional outcome and intelligence quotient. PMID- 15876755 TI - Cardiovascular response in anemia. AB - OBJECTIVE: To study the functional consequences of nutritional anemia by evaluating the exercise performance in these children. METHODS: The study was conducted on 30 each of anemic and normal children of both sexes aged between 7 and 14 years. Relevant history was taken, and detailed examination was done. These children were classified according to severity of anemia based on hemoglobin estimation as mild (10-11.9 g/dl), moderate (7-9.9 g/dl) and normal (>12 g/dl). Children with severe anemia (Hb < 7g/dl), heart disease, non nutritional anemia, acute febrile or respiratory illness were excluded. The subjects were tested on Mortara X Scribe colour stress treadmill using the modified Bruce protocol. Continuous computerised electrocardiographic analysis was done. The parameters studied included heart rate, systolic blood pressure, double product (DP = HRxSBP), ECG changes, metabolic equivalents (METS) and exercise duration. The end point of the test was a HR of 170/min (non-fatigue group) or inability to perform further (fatigue group). Statistical analysis was done by appropriate tests. RESULTS: Fifteen children each with mild and moderate anemia and 30 age-matched controls were enrolled in the study. There was no significant difference in the resting HR, exercise duration or DP between the cases and controls. Thirty-one children reached the target HR (non-fatigue group). The end-point HR was significantly lesser among the anemic children in the fatigue group (P 0.04). The percent gain of SBP, at peak exercise (P 0.0007) and recovery SBP as percent of resting SBP (p 0.006) were significantly more in the anemic children, more so in the mildly anemic ones. Lesser METS was achieved by anemics as compared to controls (P 0.04). ECG changes occurred significantly more often in anemic children, 53.8% of those with changes being moderately anemic, the main abnormality being ST depression. None developed arrhythmia during exercise testing. CONCLUSION: The cardiovascular response to physical exercise is compromised in children with nutritional anemia and hence these children may never attain their full potential in various school activities. Prevention of anemia should be a priority in school going children. PMID- 15876756 TI - Impact of availability of oral hydrocortisone on growth of children with CAH. AB - OBJECTIVES: 1. To compare growth parameters of patients with Congenital Adrenal Hyperplasia (CAH) managed on Prednisolone (PR) before and on Hydrocortisone (HC) after its availability in India. 2. To compare growth parameters of patients with CAH who have been on treatment with HC since diagnosis with patients managed on PR. METHODS: Growth parameters of twelve children (8 m, 4 f) with congenital adrenal hyperplasia were retrospectively studied while on treatment with prednisolone (PR) earlier and then hydrocortisone (HC) after it became freely available in India. RESULTS: Patients treated with PR had height Z score of 0.42, weight Z score of - 0.45, and height velocity Z score of -2.06. On HC these scores were -0.27, +0.16 and + 2.27. Patients treated with HC from the begining had a height Z Score of + 0.08, weight Z score of +0.22, and height velocity Z score of +0.68. CONCLUSION: Hydrocortisone has a less growth effect than prednisolone and patients treated with HC from the beginning showed near normal growth. PMID- 15876759 TI - Pediatric epilepsy -- an Indian perspective. AB - Prevalence studies from India suggest that epilepsy prevalence is similar to developed nations. Neurocysticercosis (NCC) predominates as an etiology. A large treatment gap is still a public health problem. Benign epilepsies and West syndrome appear to be underrepresented in studies on classification of seizures/syndromes. Febrile seizures prevalence in India is similar to other countries and appear to be as benign. Risk factors of intractable epilepsy (IE) in Indian studies include early age of onset, neurodevelopmental abnormalities and certain seizure types. Perinatal injuries underlie many IE. Many IE are not truly intractable and respond to simple therapeutic measures. The ketogenic diet and surgery are other methods now being used in Indian centers. Neurocysticercosis and neonatal hypoglycemic brain injury, two widely prevalent etiologies are reviewed in detail. PMID- 15876760 TI - Leukodystrophies: Indian scenario. AB - The leukodystrophies are familial disorders with onset usually in infancy or childhood. The clinical features consist of motor dysfunction with varying degree of cognitive decline. Magnetic Resonance Imaging (MRI) has helped to identify and characterize these disorders. In some leukodystrophies, biochemical enzymatic and genetic defects have been identified. The commonest leukodystrophy seen in India is Megalencephalic Leukodystrophy with subcortical cysts. The essential features consist of large head, mild pyramidal and cerebellar dysfunction, and occasional seizures. MRI studies show extensive white matter changes with temporal cysts. It is common in the Agarwal community in India. An identical mutation in exon 2 of the MLC 1 gene has been identified in this community suggesting a founder effect. PMID- 15876761 TI - Thalassemia and related hemoglobinopathies. AB - Hemoglobinopathies are the most common single gene disorders in man. There are several hundred of these disorders though the thalassemias -- alpha and beta and the sickling disorders make up the vast majority. Recent advances in the understanding of the hemoglobin structure and the genetics of its synthesis has contributed significantly to the understanding of these diseases. Disorders include those with reduced globin synthesis, abnormal globin chains and failure to switch globin chain synthesis at the appropriate age. This review focuses on the clinical features, diagnosis and management strategies of the alpha and beta thalassemias, the sickling disorders and touches on a few rarer hemoglobinopathies. It also emphasizes prevention strategies and chronic transfusion safety in countries like India where there are limited resources. PMID- 15876762 TI - Inborn errors of metabolism (IEM) -- an Indian perspective. AB - The inborn errors of metabolism (IEM) constitute a diverse heterogeneous group of disorders with protean clinical manifestations presenting mainly in the pediatric population. Though individually rare, together they constitute a significant percentage of children seen in genetic and neurology clinics. This review focuses on selected IEMs and highlights those seen in the neonatal period. Data from Indian centers are presented. It also emphasizes principles of management in these difficult disorders in the context of a developing country. PMID- 15876763 TI - Tuberculosis -- persistent threat to human health. AB - With the increasing incidence of tuberculosis worldwide, childhood cases now constitute 40% of the total. TB control thus has global importance. Unfortunately, control of disease is not in sight. It was always thought that adult tuberculosis is the fountainhead of childhood tuberculosis but it is being increasingly realized that it is the infection acquired during childhood that promotes reactivation of adult disease, which in turn maintains the chain of transmission. Thus childhood tuberculosis needs equal or more attention for effective control. Early detection by simple tests and ensuring treatment compliance is the goal. The small number of bacilli and inaccessible sites for bacteriological confirmation makes diagnosis of childhood tuberculosis difficult. Circumstantial evidence is often the basis of diagnosis. However, as clinical manifestations depend upon host immune response and virulence of tubercle bacilli, there is no typical clinical presentation. A large number of infected children may remain asymptomatic, undiagnosed and untreated. Conventional tests such as tuberculin test and radiology are not fully dependable and newer tests have limitations. Poor patient treatment compliance contributes to failure of a tuberculosis control program and leads to drug resistance. To combat this, direct observed treatment (DOTS) has been unanimously recommended in treatment of tuberculosis. DOTS is however estimated to be used in less than 40% of new cases. Misconceptions threaten to undermine continued success in tuberculosis control. TB control is essentially a management problem. Greater accountability of governments, donors and providers is essential. PMID- 15876764 TI - Ethical issues in pediatric intensive care in developing countries: combining western technology and eastern wisdom. AB - Application of traditional ethical principles in developing countries may not, indeed should not, conform to the western philosophy and ideology. The principle of distributive justice is of utmost importance when critical resources are scarce. There is no ethical imperative, nor is one followed even in the most advanced countries, that every citizen is entitled to the very best available care. However, a society must establish a uniform code of ethics that can be applied nationally, whereby all citizens are eligible for a minimum acceptable level of care. The traditional principles of autonomy, beneficence, nonmaleficence and justice are still applicable in structuring an ethical framework that is most suited for the country's needs and resources. PMID- 15876765 TI - Autistic spectrum disorders. AB - Autistic spectrum disorders is a complex developmental disorder with social and communication dysfunction at its core. It has a wide clinical spectrum with a common triad of impairments -- social communication, social interaction and social imagination. Even mild or subtle difficulties can have a profound and devastating impact on the child. To be able to provide suitable treatments and interventions the distinctive way of thinking and learning of autistic children has to be understood. The core areas of social, emotional, communication and language deficits have to be addressed at all levels of functioning. The important goals of assessment include a categorical diagnosis of autism that looks at differential diagnosis, a refined precise documentation of the child's functioning in various developmental domains and ascertaining presence of co morbid conditions. The interventions have to be adapted to the individual's chronological age, developmental phase and level of functioning. The strategies of curriculum delivery and teaching the child with autism is distinctive and includes presence of structure to increase predictability and strategies to reduce arousal of anxiety. PMID- 15876766 TI - Wolman disease: diagnosis by leucocyte acid lipase estimation. AB - Wolman disease is a rare fatal autosomal recessive disorder caused by absence of acid lipase enzyme leading to accumulation of cholesterol ester. Hepatosplenomegaly is a constant feature and occurs as early as fourth day of life. Progressive mental deterioration may occur after few weeks of onset of symptoms. Adrenal calcification seen on X-ray abdomen, USG or CT scan is the hallmark of Wolman disease. For the first time in Indian literature, the authors report a case of Wolman disease that was confirmed by acid lipase enzyme estimation. PMID- 15876767 TI - Fibrochondrogenesis. AB - Fibrochondrogenesis is a rare lethal short-limb skeletal dysplasia. Till now only fifteen cases have been reported since Lazzaroni-Fossati first described it in 1978. Hence reported a case of fibrochondrogenesis in a child born to a consanguineously married couple with characteristic physical and radiological features and discuss the incidence, inheritance, ultrasonographic, clinical, radiological and pathological characteristics of this disorder. PMID- 15876768 TI - Aplastic anemia in an HIV infected child. AB - Hematologic manifestations of HIV in children are common and include anemia, neutropenia, lymphocytopenia, thrombocytopenia that may occur due to many reasons. However, aplastic anemia due to HIV infection is rare and even more so in children. Though anemia is seen with advanced disease and associated with poor prognosis it is treated with various therapeutic modalities. Our patient with aplastic anemia due to HIV infection responded to antiretroviral therapy. PMID- 15876769 TI - Bronchogenic cyst -- cause of refractory wheezing in infancy. AB - A 1 1/2-year-old male child with past history of recurrent wheezing presented with a severe and refractory episode following a peanut aspiration. A bronchoscopy was done and the peanut removed. However, as the wheezing persisted a detailed evaluation was done which revealed a bronchogenic cyst. Surgical excision of the cyst was done and the child improved symptomatically. Congenital lesions need to be ruled out in infants and children with persistent and refractory wheezing. PMID- 15876770 TI - Labial adhesions: facts and fiction. PMID- 15876772 TI - Arthritis in HIV. PMID- 15876773 TI - Silent blast in oral cavity: is the car battery innocuous? AB - Low voltage energy source is not free from danger. An exceptionally rare and peculiar mode of facial blast injury is reported. The blast took place silently in the mouth of a 15-year-old boy, due to short-circuiting of wires connected to a12-volt car battery while peeling off insulation with the intent of running a musical instrument. Airway compromise due to soft tissue injury produced further problem during tracheostomy. Emergent airway management is discussed. Post healing sequel resulted in loss of voice and prevented normal oral feeding. The case report emphasizes need for education to public in handling low voltage energy source. PMID- 15876774 TI - Teratoma of palatine tonsil in a neonate. AB - Among many causes of respiratory distress and feeding difficulties in the neonatal period, a tumor involving oropharyngeal region is very rare. But still rarer is the unique presentation of this case of teratoma tonsil with intermittent attacks of choking with cyanosis. PMID- 15876775 TI - Sirenomelia apus with vestigial tail. AB - Sirenomelia is an exceptionally rare congenital malformation characterized by complete or near complete fusion of lower limbs. A newborn with clinical features of sirenomelia including fused lower limbs in medial position, absent fibula, anal atresia, complete absence of urogenital system (bilateral renal agenesis, absent ureters, urinary bladder, absent internal and external genitalia), a single umbilical artery and a vestigial tail is reported. Association of vestigial tail with sirenomelia is not described in the literature. PMID- 15876776 TI - Thoracic three-dimensional spiral CT findings of an infant with spondylothoracic dysostosis. AB - Spondylocostal dysostoses are a group of rare inherited disease with a heterogeneous disorder of vertebral segmentation defects and rib anomalies, which lead to respiratory problems predicting the clinical outcome. Spiral CT with three-dimensional (3D) imaging provides exact measurement of the bony rib cage. We report a case of an infant with spondylothoracic dysostosis, a phenotype of spondylocostal dysostoses, and 3D spiral CT findings of his rib cage since it may contribute to the surgical planning. PMID- 15876777 TI - Yolk sac tumor of vagina. AB - Yolk sac tumors are common in children. By virtue of being chemosensitive, they are amenable to cure by chemotherapy alone and radical surgery is often not required. Yolk sac tumors occurring in the vagina are rare and thus may not be recognized early or may be inadvertently subjected to radical surgery. The authors report a case that presented to them after radical surgery with elevated Alpha-fetoprotein level is reported. The management of this case and review of the relevant literature are discussed here. PMID- 15876778 TI - Contraceptive practices and awareness of emergency contraception in educated working women. AB - BACKGROUND: Regular contraceptive use and emergency contraception are tools to prevent pregnancies. AIMS: This study was designed to investigate knowledge and use of contraceptive methods and awareness of emergency contraception among women working in the hospital. SETTINGS: Educated workingwomen in a medical college hospital. DESIGN: Cross-sectional study. MATERIALS AND METHODS: The study was carried out among women belonging to three categories: staff nurses, ministerial staff and others. Married as well as unmarried employees in the reproductive age group were interviewed. A pretested mixed questionnaire containing open as well as closed ended questions was administered. The women were asked questions concerning knowledge and use of contraceptive methods and awareness of emergency or postcoital contraception. RESULTS: Of the 284 employees 258 women consented for the interview. All the subjects were literate and majority (97.2%) had an urban background. Of the 190 married women, 154 (81.1%) practiced contraception, among them (73.3%) were regular users. Eighty respondents underwent abortions of which 46 had spontaneous and 34 had induced abortions. Among the available contraceptive methods, condom was the most popular method in 89 (57.8%) followed by Copper T in 38 women (24.7%). The use of hormonal contraception was very low 2.6%. Print and electronic media were the common source of public awareness in 149 subjects (57.7%). Twenty-nine women (11.2%) were aware and only three women used emergency contraception. CONCLUSIONS: A high percentage of females in this literate workingwomen population used contraception; however, the awareness of emergency contraception was low. PMID- 15876779 TI - Molecular analysis of fragile X syndrome in Antalya Province. AB - BACKGROUND: Detection of the (CGG)n repeats in the FMR1 gene that cause the fragile X syndrome (FXS), has become a milestone for phenotype-genotype correlation in FXS. AIMS: To screen the FMR1 gene CGG repeats in index cases with FXS and their family members in the Antalya Province. SETTING AND DESIGN: This study was prospectively conducted between January 2000 and March 2005 in Department of Medical Biology and Genetics, Faculty of Medicine, Akdeniz University, Antalya. MATERIALS AND METHODS: A series of 132 cases from three hospitals in Antalya Province were studied. All cases were molecularly screened using non-radioactive Expand Long PCR method that was confirmed by Southern blotting. RESULTS: Seventeen out of 132 cases were found to have a full mutation, including three that were mosaic for premutations/full mutations. Of the 132 cases, eight were found to have the premutation size of the CGG repeats. The remaining 107 cases were identified as normal. CONCLUSIONS: Due to premature ovarian failure and Fragile X premutation Tremor/Ataxia Syndrome related with the premutation, the detection of the premutation will provide valuable information both for clinical follow-up and genetic counseling. In conclusion, our data suggest that expansion of CGG repeats in the FMR1 gene can be analyzed by Expand Long PCR, an efficient and non-radioactive method that can be used to monitor the expansion of premutation to full mutation, which would eventually lead to reduce the FXS prevalence. PMID- 15876780 TI - Ethnobotanical study of medicinal plants used by the Jah Hut peoples in Malaysia. AB - CONTEXT: An ethnobotanical study was carried out among the Jah Hut people who live in the central part of peninsular Malaysia. MATERIALS AND METHODS: The information on the medicinal plants was obtained from interview with a traditional medicinal man. The traditional uses and remedies were documented. The literature searches were carried out for the evaluation on the current status of investigations on these plants. RESULTS: In this study, we present 16 species of plants, which are commonly used among the Jah Hut people to cure some common diseases. DISCUSSION: This study is important to preserve the knowledge of medicinal plants used by Jah Hut people. The surveys of phytopharmacological literatures of these plants have great pharmacological and ethnobotanical significance. PMID- 15876781 TI - Eosinophilia and valvular heart disease. PMID- 15876782 TI - Olanzapine-induced double incontinence. PMID- 15876783 TI - Pamidronate--a promising new candidate for the management of spondyloarthropathy. AB - Bisphosphonate group of agents are known for their anti-bone resorptive properties. However, recently their anti-inflammatory and anti-arthritis properties have come to light. Clinical trials of their use in spondyloarthropathy are showing promising results, especially in patients with shorter disease duration. The adverse event profile is mainly limited to postinfusion arthralgia, myalgia and fever. The concept of pamidronate in spondyloarthropathy management should be evaluated further in light of these clinical studies and could have a major impact on our resource-restricted setting. PMID- 15876784 TI - An experimental study of apico-aortic valved conduit (AAVC) for surgical treatment of aortic stenosis in dogs. AB - A new valved conduit was developed using a canine aortic valve. The bioprosthetic valve was fixed with glutaraldehyde and epoxy compound (Denacol-EX313/810). A vascular graft composed of ultra-fine polyester fiber (10 mm in diameter, 200 mm in length) was used. Four dogs underwent apico-aortic valved conduit (AAVC) implantation and aortic banding (bypass group, BG), while another 4 dogs underwent aortic banding without AAVC implantation (control group, CG). Cardiac catheterization and angiocardiography were performed for assessment of hemodynamics 2 weeks and 6 months after surgery. Left ventricular systolic pressure, left ventricular end-diastolic pressure and the left ventricular-aortic pressure gradient differed significantly (P<0.01) between the BG and CG dogs. Left ventricular angiocardiography showed patency of the valved conduit in all the BG dogs. Echocardiography was performed before and 2, 4 and 6 months after surgery, and showed that while pressure overload caused concentric myocardial hypertrophy in the CG dogs, the left ventricle dilated eccentrically in the BG dogs. Furthermore, relief of left ventricular pressure overload by AAVC was maintained. PMID- 15876785 TI - Proliferative responses to canine thyroglobulin of peripheral blood mononuclear cells from hypothyroid dogs. AB - The immune responses of hypothyroid dogs to canine thyroglobulin (cTg) were evaluated for the proliferative ability of peripheral blood mononuclear cells (PBMC). PBMC from three hypothyroid dogs with high titers of thyroglobulin autoantibody (TgAA) and 3 clinically normal dogs were cultured with 5, 10, or 20 microg/ml of cTg for 72 hr. The proliferative responses of the cells were determined by the level of incorporated BrdU. The numbers of cells expressing Thy 1, CD4, CD8 and IgG in the PBMC were counted by the immunofluorescence method. Proliferative responses to cTg were observed in the cells from hypothyroid dogs. The number of cells expressing IgG and CD8 in the hypothyroid dogs tended to be high compared with the clinically normal dogs. The CD4+ cells in cultures from hypothyroid dogs increased depending upon the amount of cTg. There was a significant (P<0.05) positive correlation between the number of CD4+ cells and the concentration of cTg in the cultures from hypothyroid dogs. These findings suggest a possible relationship between canine hypothyroidism and cellular immunity. Loss of self tolerance to thyroid antigens in CD4+ T cells may play an important role in the development of canine hypothyroidism. PMID- 15876786 TI - Differential invasion by Sendai virus of abdominal parenchymal organs and brain tissues in cortisone- and cyclophosphamide-based immunosuppressed mice. AB - Pneumotropic virus invasion of parenchymal organs in mice with immunosuppressants is worth studying from an etiopathogenetic viewpoint as an animal model of a compromised host. This study analyzed the invasion by Sendai virus (SeV) of mouse organs in immunosuppressive states induced by cortisone (CO) and cyclophosphamide (CY). After subcutaneous inoculation of CO or CY, or both, in mice infected intra nasally with SeV, the SeV invasion was investigated by detecting viremia and viral antigen in organs. SeV Nagoya strain and one treatment of CO or CY caused viremia at 18 hr or 3 d, respectively, with infection in tracheal lymph nodes, but not in untreated mice. SeV invaded hepatocytes and splenolymphocytes on days 10 and 5-10, respectively, after infection. CO or CY treatment, three times, induced viral invasion of brain tissues or serious respiratory infection, respectively, but progressive invasion of abdominal parenchymal organs was not different between treated groups. One CO and two CY treatments intensified viral invasion into many organs, maintaining seronegativity. Cerebral blood vessels had the highest incidence of viral antigen in the brain. Astroglial sheets, choroid plexuses, pia maters, and ventricular epithelia tested positive; test-positive neurons were few. SeV MN strain caused progressive invasion of the brain with gliosis and neuronophagy. Blood-brain barrier disruption was caused by virulence of the MN strain. Half the infected mice in two groups treated with CO once and CY twice succumbed to delayed hypersensitivity, suggested by cerebro microvascular nodulation. PMID- 15876787 TI - Anesthetic and cardiovascular effects of balanced anesthesia using constant rate infusion of midazolam-ketamine-medetomidine with inhalation of oxygen-sevoflurane (MKM-OS anesthesia) in horses. AB - The anesthetic sparring and cardiovascular effects produced by midazolam 0.8 mg/ml-ketamine 40 mg/ml-medetomidine 0.05 mg/ml (0.025 ml/kg/hr) drug infusion during sevoflurane in oxygen (MKM-OS) anesthesia was determined in healthy horses. The anesthetic sparring effects of MKM-OS were assessed in 6 healthy thoroughbred horses in which the right carotid artery was surgically relocated to a subcutaneous position. All horses were intubated and ventilated with oxygen using intermittent positive pressure ventilation (IPPV). The end-tidal concentration of sevoflurane (ET(SEV)) required to maintain surgical anesthesia was approximately 1.7%. Heart rate and mean arterial blood pressure averaged 23 41 beats/min and 70-112 mmHg, respectively. All horses stood between 23-44 min after the cessation of all anesthetic drugs. The cardiovascular effects of MKM-OS anesthesia were evaluated in 5 healthy thoroughbred horses ventilated using IPPV. Anesthesia was maintained for 4 hr at an ET(SEV) of 1.7%. Each horse was studied during left lateral (LR) and dorsal recumbency (DR) with a minimum interval between evaluations of 1 month. Cardiac output and cardiac index were maintained between 70-80% of baseline values during LR and 65-70% of baseline values during DR. Stroke volume was maintained between 75-85% of baseline values during LR and 60-70% of baseline values during DR. Systemic vascular resistance was not different from baseline values regardless of position. MKM-OS anesthesia may be useful for prolonged equine surgery because of its minimal cardiovascular depression in both of lateral and dorsal recumbency. PMID- 15876788 TI - Molecular cloning of canine membrane-anchored inhibitor of matrix metalloproteinase, RECK. AB - The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene is one of the endogenous matrix metalloproteinase (MMP) inhibitors. It was reported that decreased RECK expression closely correlated with tumor malignancy. We determined the cDNA sequence of the canine RECK gene. The cDNA sequence and deduced amino acid of canine RECK were 2,913 bases and 971 residues, respectively. The predicted amino acid sequence of the protein showed 95.5% and 91.9% homology with human and mouse RECK, respectively. RECK mRNA expression was analyzed in various canine tissues and tumor cell lines by quantitative RT-PCR. The highest RECK expression was detected in lung and testis. In comparison with the tissues, a remarkably low expression level was detected in tumor cell lines. In addition, the RECK gene was transfected in the canine transitional cell carcinoma, and its influence on cell proliferation, migration, and invasion was analyzed. The transfected RECK gene suppressed only canine tumor invasion. These results showed that RECK might play an important role in tumor malignancy in dogs as well as in other mammalians. PMID- 15876789 TI - Localization of insulin-like growth factor I (IGF-I) in the chicken liver after fasting and refeeding: demonstration by using antigen retrieval immunohistochemistry. AB - Immunohistochemical localization of insulin-like growth factor-I (IGF-I) was investigated in the liver of fasted and refed chickens by using an antigen retrieval method. The present study is the first one showing the localization of IGF-I in the chicken liver. Immunoreactivity for IGF-I was detected on the paraffin sections of livers from the fed and refed chickens after the treatment with the antigen retrieval agent. A moderate number of cells showing IGF-I immunoreactivity were scattered in the parenchyma of the liver from fed chickens. These cells were relatively large and polygonal in shape and seemed to be hepatocytes. Reaction products were observed as a granular structure in the cytoplasm of IGF-I-immunoreactive hepatocytes. The number of immunoreactive hepatocytes was increased in the liver from refed chickens compared with fed chickens. Diffuse reaction products as well as granular ones were observed throughout the cytoplasm of IGF-I-immunoreactive hepatocytes of livers from refed chickens. There are, however, no regular patterns of the distribution of immunoreactive hepatocytes in the parenchyma of both fed and refed chickens. In the liver of the fasted chickens, clear immunoreactivity for the peptide was not observed. These data show that IGF-I is located in the chicken hepatocytes and influenced by the nutriture. PMID- 15876790 TI - Serum growth hormone and insulin-like growth factor-1 concentrations in Japanese black cattle with renal tubular dysplasia. AB - To elucidate the cause of growth retardation, 14 cattle with renal tubular dysplasia (RTD) were investigated by determining serum growth hormone (GH) and insulin-like growth factor (IGF)-1 concentrations. Compared with 6 healthy cattle, the baseline, maximum, area under the curve and peak amplitude of serum GH concentrations were higher or tended to be higher. Serum IGF-1 concentrations were lower normal or lower in cattle with RTD than in healthy cattle. Serum IGF-1 concentrations correlated significantly with body weight and chest girth, but not with serum GH and creatinine concentrations. Growth retardation in RTD might be associated partially with GH resistance. PMID- 15876791 TI - Protective effects of bio-active ceramic water on alcohol-induced hepatic injury in pigs. AB - Among many detrimental injuries, alcohol is implicated in hepatitis, fatty liver, hepatic fibrosis, and cirrhosis. The purpose of this study was to evaluate the protective effect of bio-active ceramic water on alcohol-induced hepatic injury in pigs. Twelve male Landrace pigs were divided into 3 groups. Groups 1, 2, and 3 were fed with bio-active ceramic water + normal liquid diet, bio-active ceramic water + liquid diet containing 15% ethanol, and tap water + liquid diet containing 15% ethanol for 12 weeks, respectively. For serological, histopathological, and immunohistochemical analysis, all pigs were sacrificed at week 12. In group 3, serum ALT and AST levels increased, and mild fatty change and moderate necrosis were detected in the liver. Collagen fibers, myofibroblasts, and CYP2E1 were also increased or activated in group 3. In group 2, there were mild hepatic injuries compared to group 3. However, injuries and activations were not observed in the liver in group 1. We suggest that the bio active ceramic water used in the present study had protective capability against ethanol-induced hepatic injury and that having no toxic effect on the pig liver. The bio-active ceramic water might be useful as a therapeutic drinking water in patients suffering from alcoholic liver diseases. PMID- 15876792 TI - Bovine ephemeral fever in Taiwan (2001-2002). AB - Bovine ephemeral fever (BEF), a vector-borne disease of cattle, is caused by the Ephemerovirus of the family Rhabdoviridae. In the past 40 years, Taiwan has had seven BEF epizootics, and we have previously reported the first five. This study summarizes the 2001 and 2002 epizootics; conducted case-control serologic studies on 10 herds involved in the 2001 epizootic; determined whether the recent BEF viruses have varied significantly; and discusses the relationship between epizootic patterns and possible variant BEF viruses. For mature cows that had received at least 2 doses of vaccine before the study, a negative correlation between the prevaccinated (the 3rd dose and after) serum neutralization antibody (SNA) titers and their postvaccinated peak rates was found. When prevaccinated SNA levels were at < or = 32, their postvaccinated SNA levels increased significantly faster (P<0.01) than for those at > or = 32. The glycoprotein gene of isolates from 1999, 2001, and 2002 had a 99.2-99.9% homology, without consistent amino acid variations in the neutralization sites. Phylogenetic analysis of Taiwanese isolates revealed 2 distinct clusters, the 1983-1989 and 1996-2002 isolates. Cross-neutralization tests confirmed the glycoprotein gene sequence analysis results. In conclusion, annual boosters at SNA levels > 32, at more than 2 doses, or at intervals shorter than 6 months are not advisable. The occurrence of frequent small epizootics implies the dominance of BEF virus over host immunity, but not a variant virus. PMID- 15876793 TI - Carcinogenicity of biphenyl in mice by two years feeding. AB - Carcinogenicity and chronic toxicity of biphenyl was examined in the male and female BDF1 mice fed a diet containing biphenyl at 667, 2,000 or 6,000 ppm for 2 years. There was no difference in survival rate between any biphenyl-containing diet-fed group of either sex and the respective control. Body weights of the males and females fed 6,000 ppm diet were significantly lower than the respective control. Incidences of hepatocellular carcinomas and hepatocellular adenomas in the females fed diets containing biphenyl were significantly increased in a dose related manner, and exceeded a range of the historical control data in the Japan Bioassay Research Center. Incidences of basophilic cell foci in the liver were increased in the females fed 2,000 and 6,000 ppm diets. There was no increase in tumor or tumor-related lesion in the males fed diets containing biphenyl. Chronic toxicity of biphenyl was characterized by increased incidences of urothelial desquamation in the renal pelvis in males and females and mineralization in the inner stripe of renal outer medulla in females, as well as changes in serum levels of BUN, ALP and some electrolytes in males and females. In conclusion, the 2-year oral administration of biphenyl-containing diets induced pre-neoplastic and neoplastic lesions in the liver of females and non-neoplastic lesions in the kidney of males and females. Causative factors for the biphenyl-induced hepatocarcinogenicity were discussed in light of our published finding of peroxisome proliferation. PMID- 15876794 TI - Immunohistochemical analysis of expression patterns of tumor necrosis factor receptors on lymphoma cells in enzootic bovine leukosis. AB - Tumor necrosis factor-alpha (TNF-alpha) has been reported to be associated with the progression of lymphoproliferative neoplastic diseases and retroviral infections. Hence we examined immunohistochemically the expression patterns of TNF-receptors (TNF-RI and RII) on lymphoma cells derived from the 29 cases of enzootic bovine leukosis (EBL). Lymphomas obtained in 29 animals with EBL were histopathologically classified into three types: diffuse mixed type (10 cases), diffuse large type (9 cases), and diffuse large cleaved type (10 cases). Immunohistochemically using a monoclonal antibody to a bovine lymphocyte surface antigen, the lymphomas were classified into three phenotypes: B-1a (CD5+/CD11b+), B-1b (CD5-/CD11b+) and B-2 (conventional B) (CD5-/CD11b-). Interestingly, the lymphoma cells in all animals expressed TNF-RII, but not TNF-RI. Although, in EBL, lymphoma cells of which the histopathological and immunological property differs has been formed, the expression patterns of TNF-Rs had the universality in all lymphoma cells. TNF-RII, which induces cell proliferation, was expressed but TNF-RI, which induces cell apoptosis was not expressed on all lymphoma cells, suggesting that TNF-Rs play an important role in the malignant proliferation of B cells and formation of lymphomas in EBL. PMID- 15876795 TI - The effects of transcranial electrical stimulation on anaesthesia and analgesia in rats. AB - In this study, we determined the effects of transcranial electrical stimulation (TCES) on the anaesthetic requirements of thiopental and the analgesic requirements of remifentanil, in rats. The experiments were performed on 120 albino male Wistar rats, which were randomly allocated to four groups (n=30). (Thiopental, Thiopental+TCES, Thiopental+Remifentanil, and Thiopental+Remifentanil+TCES). Animals were anaesthetized with thiopental, and 15 min later, remifentanil was injected to rats in the Remifentanil groups. TCES was started in the stimulated groups 20 min after thiopental administration. Rats were stimulated 5 times for this experiment. The times for recovery, herein called Cognition Recovery Time and Motion Recovery Time were measured. Cognition Recovery and Motion Recovery Times were not affected by the stimulation. Analgesia was assessed using the wet tail-flick latency (TFL). In the Thiopental group, the analgesia level returned to control values on the 35th min. In the Thiopental+Remifentanil group, the analgesia level returned to control values on the 50th min. In the Thiopental+ TCES group, the analgesia level reached the peak value on the 65th min. In the Thiopental+Remifentanil+TCES group, the analgesia level reached the peak value on the 35th min and analgesia remained high during the 90 min after cessation of the stimulation. The analgesic potency for the Thiopental+Remifentanil+TCES group was increased by 30-40% when compared with the prior TFL values, 160% when compared with the control group, and 50-75% when compared with Thiopental+TCES group on the 35th min (P<0.001). In conclusion, TCES markedly decreases the anaesthetic and analgesic requirements for thiopental and remifentanil in rats. PMID- 15876796 TI - Axial correction of pes varus by transverse-opening wedge osteotomy and T-plate fixation with beta-tricalcium phosphate (beta-TCP) transplantation in dachshunds. AB - Axial correction was performed surgically in two miniature dachshunds presenting with lateral patellar dislocation and limping caused by pes varus. Pes varus had resulted from asymmetric closure of the physis of the distal tibia. Prior to surgery, osteotomy was simulated by measuring X-ray films to determine the distance required for the wedge opening. Transverse-opening wedge osteotomy was performed on the medial side of the distal tibia, and beta-tricalcium phosphate (beta-TCP) was inserted in a wedge shape into the area created by the cuneiform osteotomy. Finally, the tibia was fixed by a veterinary 1.5/2.0-mm T-plate. Both dogs were able to walk a few days after surgery, and the lateral dislocation of the patella normalized almost completely in about one month. At two months, X-ray films showed that the implant had remained in position without any dislocation, and the beta-TCP had fused with the surrounding bone. PMID- 15876797 TI - Counting absolute number of lymphocytes in quail whole blood by flow cytometry. AB - In a previous study, we reported a new method for counting quail blood cells. After quail blood cells were stained with fluorescent lipophilic dye (DiOC6(3)), absolute counts of erythrocytes, granulocytes, and monocytes were obtained by means of flow cytometry (FC). The FC method has the potential for application to avian blood cells count; however, the method was unable to distinguish between lymphocytes and thrombocytes. In the present study, we improved the FC method to obtain separate counts of lymphocytes using DiOC5(3). After quail blood cells were stained with DiOC5(3), the cells were measured with FC. Each blood cell type was distinguished by means of their typical FL-1 (green fluorescence) and SSC (side scatter). Absolute numbers of erythrocytes, granulocytes, monocytes and lymphocytes in whole blood were obtained. The improved FC analysis worked equally well with chicken (Gallus gallus) and goose (Anser cygnoides) blood. PMID- 15876798 TI - Infestivity of Demodex canis to hamster skin engrafted onto SCID mice. AB - We demonstrated that Demodex canis was transferred to skin xenografts of a dog and a hamster onto severe combined immunodeficiency mice. After the transfer of mites, the number of eggs, larvae, nymphs and adult mites per gram of canine and hamster xenografts increased, whereas no live mites were detected on murine allograft. These results indicate that D. canis proliferates in hair follicles of dog and hamster skins but not in murine allograft. Therefore, D. canis may have host preference but not strict host-specificity. PMID- 15876799 TI - Detection of Lawsonia intracellularis using immunomagnetic beads and ATP bioluminescence. AB - Lawsonia intracellularis is an obligate intracellular pathogenic bacterium that causes proliferative enteropathy in various animals. The detection of L. intracellularis in clinical and environmental samples is necessary for the diagnosis of infection and epidemiological investigations. For the detection of L. intracellularis in fecal samples, we have developed an immunological method using immunomagnetic separation and ATP bioluminescence. Magnetic beads were coated with an anti-Lawsonia surface antigen (LsaA) antibody in order to capture the L. intracellularis in fecal samples from infected rabbits and the bacteria captured by the immunomagnetic beads were assayed by means of ATP bioluminescence. Our results showed that L. intracelluraris was detected by immunomagnetic separation of bacteria-holding magnetic beads and ATP-based bioluminescence, suggesting that our methods could be useful for the diagnosis of proliferative enteropathy. PMID- 15876800 TI - Increased expression of both constitutive and inducible forms of nitric oxide synthase in the delayed phase of acute experimental testicular torsion. AB - To elucidate the roles of both constitutive endothelial nitric oxide synthase (eNOS) and neuronal NOS (nNOS), and inducible NOS (iNOS) in acute experimental testicular torsion, the expression of iNOS and constitutive eNOS and nNOS were studied in the rat testis with ischemia/reperfusion (I/R) injury. Western blot analysis showed that all three isoforms of NOS increased significantly at 24-48 hr after I/R and declined slightly thereafter. After I/R, immunoreactivity for both iNOS and nNOS was detected, mainly in the interstitial space around damaged tubules, while germ cells in the damaged tubules were immunostained intensely for eNOS. We postulate that increased expression of the three NOS isoforms in the testis after I/R, which might generate nitric oxide, affects delayed germ cell death following I/R via paracrine or autocrine fashion. PMID- 15876801 TI - Hematological findings and antibody responses in Syrian hamster (Mesocricetus auratus) infected with Babesia microti. AB - Hematological findings during the course of infection and the antibody response in Syrian hamsters infected with Babesia microti were examined. A macrocytic hypochromic anemia with an increase of the reticulocyte count was detected as a rise in the parasitized erythrocyte rate. White blood cell counts also remarkedly increased with the increases of both neutrophils and active-shaped monocytes, and thus they particularly play an important role in eliminating the parasite. In Western blotting with the sera from the hamsters infected with B. microti, a 38 kDa protozoan antigen reacted to the early-term sera, and additionally 28, 32, and 34 kDa antigens also reacted to the medium- and latter-term, and convalescent sera. These antigens were immunodominant and the antibodies against these antigens had also important roles for inhibition of this parasite. PMID- 15876802 TI - Impact of elevated plasma total homocysteine concentration on coronary atherosclerosis in Chinese patients with acute myocardial infarction undergoing primary coronary intervention. AB - Elevated plasma total homocysteine (tHcy) has been considered to be a new risk factor for coronary atherosclerotic disease. However, the association has not been proven indisputably, and the strength of the relationship and the interaction of plasma tHcy with other conventional risk factors remain uncertain in the clinical setting of acute myocardial infarction (AMI). The aim of this study was to investigate whether an elevated plasma level of tHcy is an independent predictor of the late stage of coronary atherosclerotic lesions in Chinese patients with AMI, who are undergoing primary percutaneous coronary intervention (PCI). Plasma levels of tHcy were prospectively measured in 178 consecutive patients with ST-segment elevated AMI undergoing primary PCI. The plasma level of tHcy was also measured in 30 control subjects with normal coronary angiographic findings. The plasma level of tHcy was significantly higher in the patients with AMI than in control subjects (10.5 +/- 3.3 micromol/L versus 8.3 +/- 2.4 micromol/L, P = 0.0004). Multiple stepwise logistic regression analysis of the baseline characteristics demonstrated that smoking (P = 0.004) and creatinine level (P < 0.0001) were independent predictors of an elevated plasma level of tHcy. Moreover, an increased plasma level of tHcy (P = 0.003), female gender (P = 0.008), diabetes mellitus (P = 0.020), and the presence of previous myocardial infarction (P = 0.003) were independent predictors of the late stage of multivessel diffuse atherosclerosis (defined by > or = 2 epicardial vessels with moderate or severe diffuse atherosclerosis). In conclusion, this investigation supports the hypothesis that a raised homocysteine concentration is a strong risk factor for the late stage of diffuse coronary atherosclerosis in Chinese patients with AMI undergoing primary PCI. This result therefore raises the prospect of the need for major therapeutic research in Chinese patients. PMID- 15876803 TI - Relationship of C-reactive protein to adverse cardiovascular events in patients treated by percutaneous coronary intervention for stable angina pectoris. AB - Low-grade inflammation as detected by increased C-reactive protein (CRP) levels predicts the risk of cardiovascular events. However, there is still controversy over the mid-term predictive value of CRP in patients referred for elective percutaneous coronary revascularization (PCI) for stable angina pectoris. The aim of this study was to assess the relationship between baseline CRP level and mid term outcome of patients undergoing PCI. Two groups of patients with stable angina pectoris were prospectively studied. Group A consisted of 150 consecutive patients with a CRP level < or = 3 mg/L, and group B consisted of 150 consecutive patients with a CRP level > 3 mg/L undergoing PCI at our institution. Comparing both groups of patients, the analysis confirmed a significant difference between medians of the CRP levels (0.5 versus 8 mg/mL; P < 0.001). A higher level of CRP in group B was associated with a lower presence of male gender (P < 0.05) and history of myocardial infarction (P < 0.05). On the other hand, in group B there was higher occurrence of smoking (P < 0.001), hypertension (P < 0.05), hypertriglyceridemia (P < 0.001), and diabetes mellitus (P < 0.01). The incidence of myocardial infarction based on post-interventional release of TnI > 1.5 ng/mL reached 12% in group A and 14% in group B (P = 0.73). Analyses were repeated with adjustment for significant baseline variables, which did not change our findings. The incidence of adverse cardiovascular events during a six month follow-up was 13% in both groups (NS). Increased CRP serum prior to PCI was not associated with the risk and extent of procedure-related myocardial injury measured by TnI release and does not portend heightened cardiovascular risk at six months after percutaneous revascularization. On the other hand, a CRP level > 3 mg/L was associated with a higher occurrence of cardiovascular risk factors (smoking, hypertension, hypertriglyceridemia, and diabetes mellitus). PMID- 15876804 TI - Are the high levels of cytomegalovirus antibodies a determinant in the development of coronary artery disease? AB - In several epidemiological studies, it was suggested that a high titer of cytomegalo-virus (CMV) antibody meant CMV reactivation, and that this condition was a determinant of coronary artery disease (CAD). The purpose of this study was to investigate both the prevalence of the CMV infections in our study population and whether high CMV sero-positivity is a determinant of CAD. Blood samples from 179 (58 female, 121 male) individuals being evaluated for CAD suspicion by coronary angiography were tested for CMV seropositivity and CRP levels. Fifty-six patients had normal coronary arteries and 123 patients had CAD. Six patients did not have anti-CMV antibodies and 87 of the 173 seropositive patients had high levels of anti-CMV antibodies (> or = 8 U/mL). High CMV seropositivity (> or = 8 U/mL) was a significant CAD determinant even after adjustment for traditional CAD risk factors (odds ratio [OR] = 2.1 P = 0.04, respectively). The results indicate that the prevalence of high CMV seropositivity is an independent predictor of CAD in our study population and that our study population with CAD had a high rate of CMV infection. PMID- 15876805 TI - Effects of co-existence of coronary stenosis and the extent of coronary ectasia on the TIMI frame count in patients with coronary artery ectasia. AB - The measurement of the thrombosis in myocardial infarction (TIMI) frame count is a simple method for evaluating coronary blood flow. Although it is well known that slow coronary flow is present in patients with coronary artery ectasia (CAE), the effects of coexisting stenosis and the severity of ectatic involvement on coronory flow have not been adequately studied. Thus, we examined (1) the effect of coexistence of obstructive coronary artery disease on TIMI frame count (TFC) and (2) the relation between the severity of ectatic involvement and TFC in patients with CAE. Ninety-seven study patients with CAE were examined in two steps to determine if they were appropriate in terms of the aim of this study. In the first step, ectasias were divided into three groups: an isolated CAE group, a CAE group with coexisting nonsignificant stenosis, and CAE with coexisting significant stenosis. In the second step, ectasias were subdivided into three groups: CAE with one segment, two segments, and three segments (or diffuse) involvement. The TIMI frame counts for the right coronary artery (RCA), the left circumflex coronary artery (LCx), and the left anterior descending artery (LAD) in the ectasia group were significantly higher than that of the control group (P < 0.001, P < 0.05, P < 0.05, respectively). The presence of coexisting nonsignificant stenosis or coexisting significant stenosis in patients with CAE did not influence TFC (P > 0.05, for all). In addition, the severity of ectasia involvement, regardless of the localization of ectatic segments and the type of the affected vessel, did not change the TFC (P > 0.05, for all). These results suggest that neither the coexisting stenosis nor the extent of involvement significantly affect TFC in patients with coronary ectasia. PMID- 15876806 TI - Myocardium utilizes more oxygen and glucose during tepid blood cardioplegic infusion in arrested heart. AB - The aims of this study were to evaluate myocardial metabolic activity during tepid blood cardioplegic infusion in the arrested heart in comparison with cold blood cardioplegia and to assess the early clinical outcomes of these patients. Thirty patients undergoing first elective coronary artery bypass grafting surgery were included and randomized to two groups (T for tepid and C for cold), 15 patients in each. Myocardial protection was similar in both groups except for the reinfusion of blood cardioplegia, which was 6 degrees C in group C and 28 degrees C in group T (same temperature as the body perfusion). The route of cardioplegic reinfusion was antegrade during the first reinfusion and retrograde during the second reinfusion. In order to assess myocardial metabolic activity, myocardial oxygen consumption (MVO2), myocardial glucose uptake, and myocardial lactate and acid production were all calculated. Arterial and coronary venous blood samples were obtained from the aortic root cannula and coronary sinus. During cardioplegic re-infusions in the ischemic period, the calculated values of myocardial oxygen extraction, oxygen consumption, and glucose uptake were higher in group T than in group C (P < 0.05). This difference was observed during both antegrade and retrograde delivery of cardioplegic solution. Myocardial lactate production was greater in group C than in group T during cardioplegic reinfusion, both antegradely and retrogradely (P < 0.05). In all patients, cardiopulmonary bypass was terminated in the first attempt. The clinical outcome was similar in both groups. The results of this study indicate that globally ischemic myocardium is able to utilize more oxygen and glucose during cardioplegic re-infusions at a tepid temperature in comparison to cold. In addition, the data showed evidence of less myocardial injury and better left ventricular function throughout the critical period of recovery from global ischemia for the heart protected by tepid cardioplegia. PMID- 15876807 TI - Analysis of circulating apoptosis mediators and proinflammatory cytokines in patients with idiopathic hypertrophic cardiomyopathy: comparison between nonobstructive and dilated-phase hypertrophic cardiomyopathy. AB - We examined the plasma levels of soluble Fas (sFas) or Fas ligand (sFas-L), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) in patients with idiopathic nonobstructive (HNCM) and dilated-phase (DHCM) hypertrophic cardiomyopathy. Patients with idiopathic hypertrophic cardiomyopathy (HCM) may deteriorate to DHCM and the pathogenesis is unknown. The levels of these plasma cytokines were measured by ELISA and echocardiography was performed in 38 HNCM and 11 DHCM patients, and 10 normal subjects. The follow-up period was three years. In HNCM, TNF-alpha (43.3 +/- 45.2 versus 16.9 +/- 4.3 pg/mL) and IL-6 (65.1 +/- 86.4 versus 4.0 +/- 2.1 pg/mL) were slightly higher compared to normal subjects and sFas (3.7 +/- 1.2 versus 2.1 +/- 0.7 ng/mL) increased significantly. sFas (3.9 +/- 1.8), TNF-alpha (79.3 +/- 72.4), and IL-6 (234.1 +/- 135.2) in DHCM were significantly increased and only IL-6 was significantly different from HNCM. sFas-L (0.18 +/- 0.08 versus 0.25 +/- 0.05 ng/mL) in HNCM was significantly decreased, and the decrease was marked in DHCM (0.05 +/- 0.02). In HNCM, TNF alpha was negatively correlated with fractional shortening (r = -0.432, P = 0.0062) or positively with IL-6 (r = 0.665, P < 0.0001), while sFas-L was negatively correlated with IL-6 (r = -0.580, P < 0.0001). DHCM with high sFas had significantly higher cumulative incidences of worsening heart failure. The Fas/Fas-L system and proinflammatory cytokines may play an important role in the status of HCM and its progression to DHCM. PMID- 15876808 TI - Hyperhomocysteinemia: an additional risk factor in white coat hypertension. AB - The association between homocysteine and sustained hypertension (HT) has been studied. The aim of this study was to assess homocysteine levels in white coat hypertension (WCH) as an indicator of increased risk in the development of cardiovascular diseases. WCH was defined as clinical hypertension and a daytime ambulatory blood pressure of < 135/85 mmHg. Plasma levels of homocysteine were determined in patients with WCH, hypertension, and normotension (NT). The study group included 100 subjects, 33 with WCH (16 males, 17 females) aged 49.1 +/- 1.9; 35 sustained hypertensives (17 males,18 females) aged 48.5 +/- 1.7 and 32 normotensive control subjects (15 males, 17 females) aged 48.8 +/- 2.2. The subjects were matched for age, gender, and body mass index. Patients with a smoking habit, dyslipidemia, or diabetes mellitus were not included in the study. Homocysteine levels were analyzed by ELISA. Plasma homocysteine levels were significantly higher in the WCH group compared to the controls (12.32 +/- 1.07 versus 5.35 +/- 1.38 micromol/L; P < 0.001) and the WCH group had significantly lower homocysteine values than the hypertensives (19.03 +/- 0.76 micromol/L P < 0.001). Total cholesterol and tri-glycerides were not different among the groups. There were no statistically significant differences in urinary albumin excretion (UAE) or creatinine clearance between the three groups. Hypertensive retinopathy was observed in the WCH group, but was less severe and less frequent compared to HTs. LVMI was greater in the WCH group compared to the NTs, but significantly less than HTs. The data demonstrate that WCH is associated with high levels of homocysteine. The increase in homocysteine level in WCH is not as high as in SHT. Since an elevated plasma homocysteine level is a strong risk factor for coronary artery disease and there was target organ damage in our WCH group, we conclude that WCH should not be considered to be an innocent trait. PMID- 15876809 TI - Intravenous iron therapy as a possible risk factor for atherosclerosis in end stage renal disease. AB - Atherosclerosis is a disease of the arterial wall, with increasing wall thickness representing an early event in the progression of the disease. It has been suggested that iron overload, as assessed by increased serum ferritin concentration, may be a risk factor for atherosclerosis. The aim of this study was to investigate the relationship between the influence of intravenous (IV) iron therapy and ferritin levels and carotid intima media thickness (C-IMT) in dialysis patients. Sixty patients (51 +/- 14) years were divided into two groups according to their IMT obtained by ultrasound; group I (high risk) and group II (low risk). The parameters assessed were serum creatinine, urea, calcium, phosphorus, hemoglobin, albumin, uric acid, iron, ferritin, and lipid levels. Thirty-eight patients (88%) in group I and 5 patients (12%) in group II received IV iron therapy while 5 patients (29%) in group I and 12 patients (71%) in group II (P < 0.001) did not receive IV iron therapy. Ferritin levels were higher in group I than in group II (581 +/- 303 and 306 +/- 224) (P < 0.001). C-IMT measurements correlated with serum ferritin and with the intravenous iron dose received during the 24 months preceding the study (r = 0.315, P = 0.015; r = 0.471, P = 0.001). The findings indicate that IV iron therapy and elevated serum ferritin levels may cause an increase in the incidence of atherosclerosis. PMID- 15876810 TI - Altered patterns of gene expression specific to thoracic aortic aneurysms: microarray analysis of surgically resected specimens. AB - Changes in the expression levels of several genes have been described in aortic aneurysm specimens, however, the spectrum of diverse molecular alterations remains to be elucidated. We attempted to identify key molecules that modulate the pathogenesis of aortic aneurysm, using a complimentary DNA microarray carrying approximately 13,000 human genes. Segments of thoracic aortic aneurysms (TAA) and adjacent normal thoracic aortic tissues without aneurysmal changes (NTA) were obtained from 20 patients undergoing graft surgery. RNA obtained from five pairs of TAA and NTA samples was compared to determine aneurysm-specific alterations using microarray. Further, the expression levels of several genes of interest were verified in the remaining specimens by real-time reverse transcription-polymerase chain reaction (RT-PCR). In microarray assays, several types of the matrix metalloproteinases were upregulated as reported previously. Also, 220 genes suggested to be involved in protein degradation, inflammation, apoptosis, stress response, intracellular signaling, and other processes were significantly upregulated. Many of these genes have not been previously implicated in cardiovascular disease. The real time RT-PCR independently confirmed that the expression levels of MMP-2, MMP-9, ADAMTS-1, and caspase 4 were consistently increased in TAA. The results indicate that many genes are involved in a complicated manner in the pathogenesis of TAA. Investigation of these genes will help clarify the pathogenesis of this disease, and may lead to the discovery of novel therapeutic targets. PMID- 15876811 TI - Cibenzoline attenuates upregulation of Kv1.5 channel gene expression by experimental paroxysmal atrial fibrillation. AB - Antiarrhythmic drugs exert their effects by inhibiting the ion channels of cardiomyocytes. However, these effects could also modify the ionic environment around them, and thereby affect the expression of ion channels, leading to biochemical enhancement or attenuation of the antiarrhythmic effects. To test this hypothesis, the physiological and biochemical effects of cibenzoline were evaluated in a rapid atrial pacing model in rats. In rats with rapid atrial pacing, pretreatment with cibenzoline significantly inhibited the increases in Kv1.5 mRNA at 2 hours and immunoreactive protein at 4 hours by 35 +/- 15% and 30 +/- 10%, respectively. These effects were observed only in the rapid atrial pacing group, not in the sham-operated group. With cibenzoline pretreatment, 4 hour rapid atrial pacing resulted in significant prolongation of the atrial refractory period compared to the untreated group even after removal of cibenzoline. In contrast, the sham and rapid atrial pacing model with and without cibenzoline pretreatment showed similar acute physiological responses to cibenzoline. In conclusion, in addition to the acute physiological effects, pretreatment with cibenzoline exerted pleiotropic effects of inhibition of Kv1.5 channel upregulation by rapid pacing, implying differences in the cibenzoline effects when administered before and after onset of paroxysmal atrial fibrillation. PMID- 15876812 TI - Intramyocardial injection of fibroblast growth factor-2 plus heparin suppresses cardiac failure progression in rats with hypertensive heart disease. AB - A reduction of coronary flow reserve has been reported in patients with hypertensive heart disease (HHD), which suggests that myocardial ischemia may contribute to the progression to cardiac failure in HHD. Therefore, we evaluated whether fibroblast growth factor (FGF)-2 and/or heparin, which induce angiogenesis, may affect cardiac function in the setting of HHD. We used Dahl salt sensitive (DS) rats as an HHD model. Direct intramyocardial injection of 100 microg of FGF-2 plus 1.28 microg of heparin (n = 6), 100 microg of FGF-2 (n = 6), 1.28 microg of heparin (n = 6) or saline (n = 6) were performed in 9-week-old rats. Echocardiography was performed to evaluate cardiac function at 9, 11, and 13 weeks of age. Plasma atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) concentrations were measured at 8 and 13 weeks of age. DS rats were killed 4 weeks after myocardial injection (at 13 weeks of age), and myocardial capillary density was assessed by von Willebrand factor staining. Injection of FGF-2 plus heparin significantly decreased left ventricular end-diastolic diameter (P < 0.0001) and left ventricular end-systolic diameter (P < 0.0001), significantly improved the reduction of left ventricular fractional shortening (P = 0.0005), significantly decreased plasma ANP (P < 0.0001) and BNP (P = 0.016) concentrations, and significantly increased myocardial capillary density (P = 0.0002) compared with injection of saline. These findings indicate that intramyocardial injection of FGF-2 plus heparin suppresses the progression of cardiac failure in DS rats. FGF-2 plus heparin administration may be a new therapeutic strategy for the treatment of HHD. PMID- 15876813 TI - Seeding of recipient bone marrow cells reduces neointimal hyperplasia of de endothelialized rat aortic allograft. AB - Transplant vasculopathy is a leading cause of graft failure and a major contributor to the lack of success with small caliber vascular allografts. In this study we evaluate techniques of bone marrow cell seeding on small caliber vessels and assess the impact of this tactic on neointimal hyperplasia in de endothelialized rat aortic allografts. In a preliminary study, bone marrow cells from Lewis rats were seeded onto the chemically de-endothelialized luminal surface of the abdominal aorta of WKY rats - with or without fibrin glue. In the allograft transplantation model, de-endothelialized fresh aortic allografts of WKY rats were orthotopically transplanted into Lewis recipients either directly (n = 6) or after recipient bone marrow cell seeding (n = 6). Histological evaluation was performed at 28 days. Bone marrow cells were able to adhere to the de-endothelialized aortic wall owing to the use of fibrin glue, but were unable to do so without fibrin glue. In the de-endothelialized allograft transplantation model, recipient bone marrow seeding led to a significant reduction of the ratio of intimal to medial area (0.40 +/- 0.08 versus 0.79 +/- 0.08, P = 0.0077). Some of the seeded cells remained in the intima for 4 weeks and some infiltrated the media, expressing CD31 or alpha-SMA. The results suggest that recipient bone marrow cell seeding on de-endothelialized aortic allograft is feasible with the use of fibrin glue and that this technique reduces neointimal hyperplasia of the graft. PMID- 15876814 TI - Mural thrombus in an ectatic right coronary artery caused acute myocardial infarction at downstream coronary artery. AB - A 72-year-old woman was admitted to our institution because of sudden chest pain. Emergency coronary angiography revealed thrombotic occlusion of the distal right coronary artery. A large cylindrical thrombus was retrieved from her distal right coronary artery using a thrombus aspiration catheter. IVUS showed minimal atherosclerosis and moderate ectatic change at the proximal right coronary artery. A reconstructed IVUS image also showed that a mural thrombus with abrupt ending was still retained at the ectatic segment. Based on this evidence, coronary ectasia was thought to be the primary cause for the thrombus formation and acute myocardial infarction in this case. PMID- 15876815 TI - Single coronary artery with subsequent coursing of right coronary artery between aorta and pulmonary artery: fractional flow reserve of the anomalous artery guiding the treatment. AB - A single coronary artery with an anomalous origin between the pulmonary artery and aorta is an unusual congenital anomaly. We present a 60 year old female patient with stable angina pectoris. Her coronary angiogram revealed that the right coronary artery originated from the left main stem and coursed between the great vessels. There was 90% stenosis in the left anterior descending coronary artery. We performed a fractional flow reserve study of both the anomalous origin artery and stenosed vessel. Fractional flow reserve study of the anomalous RCA did not reveal functional ischemia. We did not refer the patient to coronary artery bypass grafting but instead performed percutaneous coronary revascularization of the LAD. The patient is alive and has been free of symptoms during the 1 year follow-up. PMID- 15876816 TI - Localized dissection of the sinus of valsalva without coronary artery involvement during percutaneous coronary intervention. AB - Dissection of the sinus of Valsalva is an extremely rare accident during percutaneous coronary intervention (PCI), but it can lead to serious complications such as dissection of the ascending aorta. We experienced a localized dissection of the right coronary cusp without coronary artery involvement that was induced by a guiding catheter during PCI in a patient with acute myocardial infarction. The localized dissection showed pooling of the contrast medium in the acute phase, but it subsided spontaneously after 12 days without any sequelae. Manipulation of the guiding catheter should be performed with great caution not only in the coronary artery but also in the sinus of Valsalva. PMID- 15876817 TI - Histopathological findings of the no-reflow phenomenon following coronary intervention for acute coronary syndrome. AB - Although no-reflow phenomenon may occur in patients that experience reperfusion after ischemia, there have been no reports describing the postmortem findings in these patients. We describe the findings of an autopsy in a 56-year-old man who experienced acute coronary syndrome with no-reflow phenomenon after coronary intervention. Macroscopic study demonstrated myocardial infarction with diffuse hemorrhage, and microscopic analysis revealed vascular damage and microembolization in the no-reflow area. In conclusion, coronary microembolization and damage to the small coronary artery may contribute to the pathogenesis of no-reflow phenomenon following coronary intervention in humans. PMID- 15876818 TI - Ventriculatrial block during atrioventricular nodal reentrant tachycardia suggesting existence of an upper common pathway. AB - Studies on the mechanisms of atrioventricular nodal reentrant tachycardia (AVNRT) have yet to clarify whether the slow and fast pathways connect directly with the atria or via an upper common pathway. Although a "final common pathway" connecting the slow and fast pathways to the proximal His bundle was thought to be part of the reentrant circuit, debate on the presence of an upper common pathway continues. We report a case of AVNRT continuing despite the occurrence of ventriculoatrial block, thus supporting the existence of an upper common pathway. PMID- 15876819 TI - Reversible renin-dependent renovascular hypertension successfully treated with percutaneous transluminal renal angioplasty and stenting. AB - A 37-year old woman was suspected of having renovascular hypertension because of recent onset severe hypertension (blood pressure 220/135 mmHg; compared to 132/65 mmHg two years earlier) and an abdominal bruit. A captopril renal scan indicated the presence of right renal artery stenosis. Additionally, a captopril plasma renin activity (PRA) provocation test showed a positive result for renovascular hypertension (baseline PRA = 291 microU/mL; 1 hour post-captopril PRA = 1444 microU/mL). Selective renal angiography demonstrated a severe critical stenotic lesion at the distal portion of the right renal artery. Blood pressure (BP) decreased to 136/80 mmHg one day after successful percutaneous transluminal renal angioplasty and stenting. Repeat renal angiography six months after the procedure revealed no evidence of in-stent restenosis. Blood pressure (BP = 137/76 mmHg) and plasma renin profile (baseline PRA = 23.8 microU/mL; 1 hour post-captopril PRA=22.3 microu/mL) also were normal six months following initial revascularization. Moreover, blood pressure (137/84 mmHg) and renin profile remained normal 2.5 years after the procedure (baseline PRA = 24.3 microU/mL; 1 hour post-captopril = 25.6 microU/mL). The results of this study have thus demonstrated a case of renin-dependent renovascular hypertension in which both the blood pressure and plasma renin activity profile normalized following successful percutaneous transluminal angioplasty and stenting. PMID- 15876820 TI - Congenital solitary kidney with renovascular hypertension diagnosed by means of captopril-enhanced renography and magnetic resonance angiography. AB - A 24 year-old woman had a congenital solitary kidney with renovascular hypertension due to fibromuscular dysplasia. She had been treated as having essential hypertension until she developed preeclampsia and HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome at 28 weeks of gestation. Plasma renin activity and captopril test results did not indicate any abnormalities. However, renography revealed captopril-induced deterioration. Magnetic resonance angiography was also useful to detect renal artery stenosis. These findings were confirmed by renal angiography. After successful percutaneous transluminal renal angioplasty, her blood pressure and the pattern of captopril renography normalized. PMID- 15876821 TI - Stress/Rest (99m)Tc-MIBI SPECT and 123I-BMIPP scintigraphy for indication of surgery with coronary artery to pulmonary artery fistula. AB - A 45-year old man was admitted to our hospital with chest pain occurring suddenly upon exercise and disappearing with rest within several minutes. A continuous murmur was heard at the upper sternum border. Conventional electrocardiography showed no evidence of myocardial ischemia. Coronary angiography and cardiac catheterization demonstrated a fistula originating from the left coronary artery to the pulmonary artery with an aneurysm 2 cm in size, and Qp/Qs 1.08. Treadmill exercise testing showed no ST-T change at the maximum heart rate of 160 beats/min. Stress/Rest (99m)Technetium-MIBI single-photon emission computed tomography (SPECT) and 123I-15-(p-iodo-phenyl)-3,R,S-methylpentadecanoic acid (BMIPP) scintigraphy were performed to evaluate myocardial ischemia and ischemia was identified at the perfusion area of the left anterior descending artery. From these results, the patient was diagnosed as having a coronary artery to pulmonary artery fistula with myocardial ischemia. Consequently, surgical treatment was chosen under cardiopulmonary bypass. The determination of a surgical indication using stress/rest (99m)Tc-MIBI SPECT and BMIPP scintigraphy is useful in cases showing normal TMT, such as this case. PMID- 15876822 TI - Hemodialysis--from early days to tomorrow. AB - The Nestor of chronic dialysis, Belding Scribner, was well aware of most problems which plague hemodialysis to this day, but the major problem we are confronted with today has emerged much later, i.e. excessive cardiovascular mortality. Apart from modulating duration and frequency of dialysis, there is currently no logistic or technological solution immediately apparent to solve this problem. Elucidating and correcting the cardiovascular risk is the most important challenge to clinical nephrology today. PMID- 15876823 TI - Mechanisms of solute transport in extracorporeal therapies. AB - Diffusion and convection are the main mechanisms involved in the membrane separation processes occurring in extracorporeal hemodialysis. Operational parameters should be optimized in hollow fiber hemodialyzers to achieve the maximal efficiency. The nature of blood which is a non Newtonian fluid, requires specific attention in the design of dialyzers to ensure that the blood compartment operates properly. Similar attention must be placed in the design of the dialysate compartment to ensure a homogeneous distribution of the fluid and to prevent blood to dialysate flow mismatch. Finally, the membrane represents the third component of the hemodialyzer. Membrane performance depends on the used biomaterial, its biocompatibility, the thickness, the hydrophilic-hydrophobic mixture, the hydraulic permeability and the number and diameter of the pores. In this setting, diffusion and convection tend to reciprocally interfere, producing a final result that depends on the prevalence of one or the other mechanism for every specific solute. PMID- 15876824 TI - Machines for hemodialysis. AB - Basic functions of hemodialysis machines are described. The paper focuses on essential treatment parameters and safety aspects. The cause of safety hazards and protective systems for amelioration of these hazards are described. With the exception of hemolysis caused by obstructions in the extracorporeal circuit and blood losses caused by user errors machine related accidents are rare. Foreseeable improvements of next generation hemodialysis machines will reduce the likelihood of accidents further. The accuracy of adjusted or monitored treatment parameters that may influence outcome (dialysate concentration, ultrafiltration, blood flow and on-line measured clearance) is discussed. PMID- 15876825 TI - Mathematical model to characterize internal filtration. AB - Convective-diffusive dialysis techniques have recently gained considerable favor. Indeed, convective fluxes through dialyzer membranes have been demonstrated to play a role in enhancing the clearance of middle-molecular-weight solutes. An interesting opportunity is given by exploiting the internal filtration (IF)/back filtration mechanism that occurs spontaneously in high-flux dialyzers, but is difficult to quantify. In view of overcoming this drawback, a semi-empirical, lumped-parameter mathematical model for characterization of IF phenomena was developed. The model considers a dialyzer as composed by N adjacent axial blocks. For each block, hydrodynamics in the blood and dialysate compartments are determined considering hydraulic resistance and calculating local filtration. Blood viscosity and oncotic pressure are calculated locally based on hematocrit and protein concentration. Resistance parameters were determined experimentally for the BS-UL (Toray Industries Inc., Tokyo, Japan) dialyzers. The set of equations describing the model, implemented into a software program, is solved using a numerical method. Simulations allow highlighting the role of device-, treatment- and patient-dependent parameters in affecting IF. Provided an extensive validation is carried out, the use of a mathematical model could be the key to make IF more understandable and its use reliable in clinical practice. PMID- 15876826 TI - Extracorporeal sensing techniques. AB - Many physiologic variables have been measured in the extracorporeal circulation by experimental systems but only a few systems have reached technical maturity for everyday application. Variables relating to cardiovascular function, which today can be measured in the extracorporeal system, are pressure, temperature, and measures of blood composition such as hematocrit, hemoglobin, and total protein concentration. While the measurement of blood composition and temperature is well established, and while the use of extracorporeal pressure information awaits further analysis for robust application, recent interest focused on continuous measurement of plasma sodium concentration which is believed to be of major importance for optimal blood treatment. However, problems with a simple, reliable, and continuous measurement of plasma sodium for everyday use have not yet been resolved. As can be seen from the growing interest in isothermic or isonatremic treatment modes which turns away from constant and profiled treatment modes without feedback control, the treatment goal is now to provide stable conditions within the patient so as to minimize interference with intrinsic physiological control mechanisms. This, however, requires valid and reliable measurement of the specific patient variables of interest. PMID- 15876827 TI - Water treatment for hemodialysis: a 2005 update. AB - Water for dialysis represents an additive risk factors to the chronic inflammatory state documented in patients on ESRD. The possibility of sustaining proinflammatory cytokines through microbial derived products, coming from dialysate or infused solutions, is enhanced by biofilm presence on piping and on water treatment system or monitor components. Spread use of reverse osmosis, loop distribution system and pre-treatment components tailored to local raw water characteristics have greatly contributed to a general improvement in final water quality. Notwithstanding these contributions literature still reports fatal accidents or significant percentage of dialysis units not complying to the water quality standards. Technological improvement lowers chemical contamination but microbial quality relays more on quality assurance programs than on technology. Optimal water quality represents part of the anti-inflammatory strategies we need to assure to our dialysis patients to improve outcome. PMID- 15876828 TI - Online convective therapies: results from a hemofiltration trial. AB - With the introduction of the on-line preparation of dialysis fluids, the hemofiltration technique, which has never had a widespread diffusion in its old version with the infusion bags, has gained a new interest. We planned a prospective, randomized, 3-year-long study comparing survival and morbidity in ultrapure bicarbonate dialysis (BD) with on-line predilution hemofiltration (HF). Since comorbidity is one of the main factors limiting survival, the study was addressed to patients with a severe degree of comorbidity. The paper presents the preliminary results of the trial. Sixty-four patients were enrolled and randomized to either BD (N = 32) or HF (N = 32). Mean age and dialysis vintage were comparable. Twenty patients died during the study, 12 in BD and 8 in HF. The relative risk of death was 11% higher in patients treated with BD compared to those in the HF group (p < 0.005). The number of hospitalisation events per single patient was lower, even though not significantly, in HF compared to BD (1.94 + 1.26 in HF vs 2.48 + 1.98 in BD, p = NS). As concerns biochemistry, apart from beta-2-microglobulin, any other substantial difference was not found during the study, though the small solute concentration was generally a little more elevated in HF than in BD. Dialysis hypotension showed a trend to decrease in both the dialysis modalities up to near half of the trial, then, during the last year, it remained quite stable in HF, while, on the contrary, it increased in the BD group. By the end of the protocol, patients in HF showed a 2.5% incidence of acute dialysis hypotension, while patients in BD had 23%. PMID- 15876829 TI - Selected lessons learned from the Dialysis Outcomes and Practice Patterns Study (DOPPS). AB - The Dialysis Outcomes and Practice Patterns Study (DOPPS) is a prospective, observational study of the relationships between hemodialysis (HD) patient outcomes and HD treatment practices. The DOPPS began in 1996 in the United States, expanding to France, Germany, Italy, Japan, Spain, and the United Kingdom in 1998-1999, and then to Australia, Belgium, Canada, New Zealand, and Sweden in 2002. More than 300 dialysis units have participated in the DOPPS since 1996, with mortality data collected from nearly 90,000HD patients and detailed longitudinal data from nearly 30,000 HD patients. Large sample size and the large treatment practice variation observed in the DOPPS--given its international scope of participation--provide strong statistical power to investigate many different HD practices. Furthermore, the detailed patient data collected in the DOPPS allow relationships to account for differences in a large number of patient characteristics. More than 55 papers have been published from the DOPPS; here we provide a summary of selected DOPPS findings regarding nutrition, mineral metabolism, anemia management, vascular access, depression, and use of multivitamins and statins. PMID- 15876830 TI - What did we learn from the HEMO Study? Implications of secondary analyses. AB - BACKGROUND: The HEMO Study was a randomized clinical trial designed to determine whether increasing hemodialysis dose above current standards, or using high-flux membranes, would improve patient outcome. The primary results of the trial showed no statistically significant effects of either dialysis dose or membrane flux on the primary outcome of mortality. METHODS: This report examines the implications of secondary analyses involving subgroups and secondary outcome measures for the overall interpretation of the trial. RESULTS AND CONCLUSIONS: The secondary analyses of the HEMO Study do not alter the conclusions of the primary analysis: In the context of conventional three times per week hemodialysis, neither the high-flux nor high-dose interventions substantially improved patient outcome compared to low-flux and standard-dose levels. However, certain secondary results from the trial are consistent with the hypothesis of subtle effects that may be magnified by more intensive therapies that extend beyond the limits of conventional three times per week dialysis. This hypothesis will be addressed by a pair of new randomized trials sponsored by the National Institute of Digestive and Kidney Disease (NIDDK), which will compare six times per week daily and nocturnal therapies with conventional three times per week dialysis. PMID- 15876831 TI - What are we expecting to learn from the MPO study? AB - High-flux membranes represented a major improvement in dialysis technique, but evidences supporting their clinical superiority over conventional low-flux dialysis are still inconclusive. Although several studies, most of which were observational, showed an association between high-flux dialysis and lower morbidity and mortality, the Hemodialysis (HEMO) study, the first large-scale randomized clinical trial specifically aimed at testing the effect of membrane permeability on patients' outcome, failed to demonstrate a statistical significant benefit of high-flux membranes on all-cause mortality. Although disappointing, these results should however be interpreted in light of some important limitations of the HEMO study, first of all the inclusion of both incident and prevalent hemodialysis patients, the exclusion of sicker patients and the allowance of dialyzer reuse. In this context, much is expected from the Membrane Permeability Outcome (MPO) study, a randomized clinical trial investigating the effect of high-flux membranes in a large population of incident hemodialysis patients across Europe. Inclusion of only incident patients, absence of severe exclusion criteria and no dialyzer reuse are all distinguishing features of this study. Analyses of the baseline data of the MPO study confirm the high burden of cardiovascular disease among incident dialysis patients, although comparison with the Dialysis Outcomes and Practice Patterns Study data provides further evidence of a positive selection of patients in clinical trials. PMID- 15876832 TI - A new initiative in nephrology: 'Kidney disease: improving global outcomes'. AB - The burden of kidney disease: Improving global outcomes. Chronic kidney disease (CKD) is a worldwide public health problem with an increasing incidence and prevalence of patients requiring replacement therapy. There is an even higher prevalence of patients in earlier stages of CKD, with adverse outcomes such as kidney failure, cardiovascular disease, and premature death. Patients at earlier stages of CKD can be detected through laboratory testing and their treatment is effective in slowing the progression to kidney failure and reducing cardiovascular events. The evidence-based care of these patients are universal and independent of their geographic location. This paper describes the need to develop a uniform and global public health approach to the worldwide epidemic of CKD. It is to this end that a new initiative Kidney Disease: Improving Global Outcomes' has been established. Some current and future activities of this initiative are described. They include among others modification of the classification of CKD, the development of guidelines on hepatitis C, the organisation of consensus conferences like on Renal Osteodystrophy, and the creation of a website allowing the comparison of the five main English language clinical practice guidelines in kidney disease worldwide. PMID- 15876833 TI - Is there a magic in long nocturnal dialysis? AB - Long 3 X 8 h/week hemodialysis (HD) has been used without modification in Tassin since 35 years with very satisfactory morbidity and mortality results. It can be performed in the day or overnight. The observed good outcome is mainly due to lower cardiovascular morbidity and mortality than usually reported in HD. This, in turn, is due to the good control of blood pressure (BP) and of serum phosphate level. The control of BP results from the strict extracellular volume normalization using an adequate ultrafiltration and a low salt diet. High doses of small and middle molecules lead to a satisfactory nutrition, correction of anemia, control of serum phosphate and potassium with minimal needs for medications. The treatment is cost-effective. It provides an optimal dialysis i.e. it corrects as perfectly as possible each abnormality of renal failure. Overnight dialysis is the most logical way of delivering long HD with the lowest possible hindrance on patient's life. Due to the change in case mix a decreasing number of patients are apt or willing to go on overnight dialysis; education to autonomy is more difficult, but the benefits are still there. PMID- 15876834 TI - Mid-dilution: the perfect balance between convection and diffusion. AB - Although hemodiafiltration (HDF) offers the advantage of increased convective clearance for middle molecules, there is still controversy as to whether reinfusion should occur pre- or post-filter. Mid-dilution hemodiafiltration (MD HDF) is a new HDF technique that uses a special dialyzer, MD190, which allows both pre- and post-reinfusion. While externally the dialyzer looks similar to conventional hemodialyzers, the internal fibers are divided into two bundles by a special annular header that first lets the blood pass through the peripheral bundle in post-dilution, mix with the reinfusion fluid at the opposite end of the dialyzer and then proceed (after pre-dilution) to the dialyzer blood exit. The dialyzer is able to support substantially higher reinfusion rates (10-12 l/h). We have compared the removal characteristics of several small solutes and larger middle-molecular-weight toxins by examining instantaneous clearance at 45 min, the dialysis reduction ratio and total mass removal (by spilling) in a three center prospective cross-over study. Twenty patients were randomized to a treatment sequence of one-week high-flux bicarbonate hemodialysis (HD) followed by MD HDF, or vice versa. The parameters evaluated included urea, creatinine, beta2-microglobulin, angiogenin, leptin, retinol-binding protein, and the effects on sodium, potassium, bicarbonate and calcium. Blood flow rates ranged between 300-450 ml/min (mean 359 +/- 44 HD, 367 +/- 35 MD HDF). The mean reinfusion for MD HDF was 166 +/-17 ml/min. MD HDF had a significantly better instantaneous clearance for urea (328 +/- 28 vs 277 +/- 40); creatinine (292 +/- 32 vs. 212 +/- 66); phosphate (324 +/- 38 vs. 242 +/- 63); beta2-microglobulin (249 +/- 27 vs. 100 +/- 24); angiogenin (173 +/- 27 vs. 28 +/- 32); and leptin (202 +/- 29 vs. 63 +/- 43). Treatments were well tolerated with no adverse reactions occurring during any of the treatments. The MD HDF filter's unique configuration is designed to deliver high-efficiency HDF with a significant improvement in small and middle molecule removal. MD HDF supports substantially higher ultrafiltration rates, and as such, results in a higher removal of middle-molecular-weight toxins. PMID- 15876835 TI - Sequential hemofiltration-hemodiafiltration technique: all in one? AB - Sequential dialysis techniques (i.e. pure ultrafiltration followed by dialysis) have been used in the past, due to their capability to remove large volumes of fluids without inducing hemodynamic instability. The disadvantages of the inadequate dialysis and the lack of technology lead to the decline such methods. Hemofiltration (HF) and hemodiafiltration (HDF) are recently being utilized in a greater proportion thanks to the on line fluid preparation systems. Each process (HF and HDF) has its own benefits in the removal of small, medium and high molecular weight substances and in the hemodynamic stability. Sequential hemofiltration/ hemodiafiltration (SHF/HDF), may combine the benefits and eliminate the disadvantages of each method. Furthermore they can be easily applied nowadays, due to the development of new high technological hemodialysis machines. In order to evaluate the feasibility and the effects of SHF/HDF we studied 7 chronic hemodialysis patients (6 months of treatment with SHF/HDF switched to 6 months of SHDF/HF), using the same machine (AK200 ULTRA), with on line fluid preparation system and the same type of dialyzer (Polyflux 210). The feasibility of such techniques (SHF/HDF or vice versa) resulted excellent. All sessions left the patients in a condition of well-being making fulltime work. No difference was observed between the different period of treatment, but a reduction in pre value was observed in calcium-phosphorous product, C-reactive protein and beta2-microglobulin, at the end of the sequential techniques. SHF/HDF therapy is a very promising technique. Further studies are needed to better explore the potential of such a therapeutic approach in the quality of life, the hemodialysis adequacy and the hemodynamic stability of our patients. PMID- 15876836 TI - Should nephrologists be in charge? AB - Once, vascular access (VA) for hemodialysis treatment was initiated by nephrologists: Scribner introduced the arteriovenous shunt, Shaldon the central venous catheters and Brescia-Cimino the arteriovenous fistula. Later on, creating VA became a domain of surgery. Many nephrologists felt out of responsibility. Interventional procedures, angioplasty and stent insertion are mostly performed by radiologists. In 2005, the role of the nephrologist in comprehensive VA care must be newly identified. We know about the value of early referral to nephrologist and access surgeon to preserve venous vasculature. The nephrologist cares for clinical examination of vessels, for an ultrasound Doppler evaluation before the creation of primary VA with clear preference to native arteriovenous fistulae to aim at an early failure rate. Surveillance and monitoring require the exclusive responsibility of the nephrologist and his team. Early diagnosis of VA dysfunction allows elective revision before the onset of thrombosis. There should be an agreement on strategies between nephrology, surgery and radiology. Surgical techniques and skills are to be assessed from time to time. Worldwide, new organizational structures in creation, control and documentation of VA are needed. Flexibility between the disciplines involved as well as educational programs for nephrologists, surgeons and radiologists are future challenges. PMID- 15876837 TI - Challenges in interventional nephrology. AB - Lately, there has been a progressive decrease in the interest of nephrology as a medical sub-specialty reflected primarily in the decreasing number of renal fellows. Rising costs in establishing and running dialysis clinics and the lost of nephrologic procedures previously performed by nephrologists are among the many reasons for this disillusionment with the specialty. The care of chronic kidney patients frequently involve many diagnostic and interventional radiological procedures such as: diagnostic renal ultrasonography, ultrasound guided kidney biopsies, placement of tunneled hemodialysis catheters or peritoneal catheters, sonographic and radiological investigation of vascular access dysfunction, etc. Most of these procedures are nowadays performed by radiologists, vascular surgeons and surgeons in general. This fragmentation does not optimize medical care and it is inconvenient to the patient. This has led many nephrologists to introduce a new paradigm in kidney patients management, often referred as interventional nephrology (IN). This new breed of nephrologists have acquired diagnostic and interventional skills for procedures usually done by others with an added clinical perspective. To train nephrologists in these procedures and avoid the fragmented care of renal patients, the American Society for Diagnostic and Interventional Nephrology was established in 2000 and the International Society of Nephrology in 2004 introduced a new committee to address the issues of IN. It is hoped that concerted efforts will help to rescue these activities for the nephrologists and improve quality of patient care. PMID- 15876838 TI - Vascular access education, planning and percutaneous interventions by nephrologists. AB - To optimize vascular access care of patients with end stage renal disease, nephrologists themselves are taking a keen interest in the management of vascular access-related issues. Because of their unique clinical perspective on dialysis access and better understanding of the intricacies of renal replacement therapy, nephrologists are ideally suited for this activity. Two areas are the main focus of attention by these specialists: vascular access education and access-related percutaneous interventions. Vascular access-related procedures commonly performed by nephrologists include percutaneous balloon angioplasty for vascular access stenosis, thrombectomy procedure for a thrombosed arteriovenous access, tunneled hemodialysis catheter-related procedures and vascular mapping to determine the patient's optimal vascular access. While the performance of these procedures by nephrologists offers many advantages, appropriate training in order to develop the necessary procedural skills is critical. Recent data have emphasized that a nephrologist can be successfully trained to become a competent interventionalist. In addition to documenting excellent outcome data, multiple reports have demonstrated safety and success when these procedures are performed by nephrologists. This chapter focuses on vascular access education and hemodialysis access-related procedures performed by nephrologist and calls for a proactive approach in optimizing this aspect of patients care. PMID- 15876839 TI - Whole body--single frequency bioimpedance. AB - BACKGROUND: The postdialysis target weight is determined as the lowest weight a patient can tolerate without intradialytic symptoms or hypotension. Patterns of electrical properties of tissues allow a direct monitoring of fluid status without the need of the body weight. METHODS: Whole body impedance is measured from skin electrodes on hand and foot. Impedance (Z vector) is a combination of resistance, R, i.e. the opposition to the flow of an injected alternating current, at any current frequency, through intra- and extracellular ionic solutions, and reactance, Xc, i.e. the dielectric component of cell membranes and organelles, and tissue interfaces. Measurements at 50 kHz current frequency are done with the best signal to noise ratio. RESULTS: Cyclical tissue hydration changes in hemodialysis patients are detectable as changes in the whole body impedance, which can be utilized with patterns of impedance vector analysis in monitoring the prescription of optimal hydration independent of the body weight. Wet-dry weight prescription based on impedance vector analysis should bring abnormal vectors back into the reference, 75% tolerance ellipse, where tissue electrical conductivity is restored. CONCLUSIONS: Identification and ranking of normal versus abnormal tissue hydration can be obtained from impedance vector patterns without the need of equations. PMID- 15876840 TI - Use of segmental multifrequency bioimpedance spectroscopy in hemodialysis. AB - Whole body bioimpedance (BI) appears to be accurate and reproducible in the assessment of body composition, but does not appear useful for estimation of dry weight. Segmental BI has been used for the assessment of muscle mass in body segments, such as arms or legs and may be useful for rehabilitation studies. A promising new development is the application of segmental BI for dry weight determination. Changes in extracellular volume of the calf are recorded continuously during HD, thereby allowing the detection of a time-point at which no further volume is removed from the calf despite ongoing ultrafiltration (UF). Continuation of UF beyond this point is associated with an increased risk of intradialytic hypotension. This new technology may help optimizing the prescription of dry weight and UF rates in hemodialysis patients. PMID- 15876841 TI - Cardiovascular risk in patients with end-stage renal disease: a potential role for advanced glycation end products. AB - Cardiovascular complications are a major cause of morbidity and mortality in patients with end-stage renal disease (ESRD). Advanced glycation end products (AGE) are elevated in the plasma of such patients and are also found in atherosclerotic plaques. The cellular signalling pathway(s) underlying AGE induced platelet aggregation have not been elucidated. One pathway currently receiving increased attention is the externalization of the membrane phospholipid, phosphatidylserine (PS), which plays an important role in the activation of clotting factors. In this study, we have investigated ex vivo a possible link between elevated AGE concentration and PS externalization. We observed (i) increased PS externalization in platelets from patients with ESRD, (ii) reconstitution of healthy platelets with serum from patients with ESRD resulted in increased PS externalization and (iii) incubation of platelets with purified human serum albumin (HSA)-AGE elicited PS externalization suggesting a role for AGE. PMID- 15876842 TI - Improving removal of protein-bound retention solutes. AB - Recent in vitro and clinical evidence suggest that protein-bound uremic retention solutes contribute substantially to the pathophysiology of the uremic syndrome. As compared to their water-soluble counterparts however, the removal of these molecules by conventional dialysis techniques is limited. It was the purpose of the present paper to review the existing data on the dialytic removal of protein bound solutes and on the potential advantages of newly developed epuration techniques. Furthermore, the toxicity profile of this group of molecules is discussed. PMID- 15876843 TI - Inflammation in end-stage renal disease--a fire that burns within. AB - Cardiovascular disease (CVD) remains the major cause of morbidity and mortality in end-stage renal disease (ESRD) patients. As traditional risk factors cannot alone explain the unacceptable high prevalence and incidence of CVD in this population, inflammation (which is interrelated to insulin resistance, oxidative stress, wasting and endothelial dysfunction) has been suggested to be a significant contributor. Indeed, several different inflammatory biomarkers, such as high sensitivity C-reactive protein (hs-CRP), have been shown to independently predict mortality in ESRD patients. As CRP is so strongly associated with vascular disease it has been suggested that this hepatic-derived protein is not only a marker, but also a mediator of vascular disease. Indeed, recent in vitro data from studies on endothelial cells, monocytes-macrophages and smooth muscle cells support a direct role for CRP in atherogenesis. The causes of the highly prevalent state of inflammation in ESRD are multiple, including decreased renal function, volume overload, comorbidity and intercurrent clinical events, factors associated with the dialysis procedure and genetic factors. Recent evidence suggests that several cytokine DNA polymorphisms may affect the inflammatory state as well as outcome in ESRD patients. As interventions directed towards traditional risk factors have, so far, not proven to be very effective, controlled studies are needed to evaluate if various pharmacological as well as non-pharmacological anti-inflammatory treatment strategies, alone or in combination, may be an option to affect the unacceptable high cardiovascular mortality rate in this patient group. PMID- 15876844 TI - Interaction between nutrition and inflammation in hemodialysis patients. AB - The excessive cardiovascular mortality of dialysis patients is at least in part related to chronic inflammation, which is associated with the occurrence of malnutrition. The negative effects of chronic inflammation on nutritional status are mediated by proinflammatory cytokines leading to a reduction in appetite and increased muscle catabolism. However, dietary behavior itself may also independently affect inflammation. Reduced dietary supply of vitamins C, B6, B12 and folate, as well as regular coffee consumption and increased intake of dietary advanced glycation end products may trigger chronic inflammation. On the other hand, a Mediterranean dietary pattern and regular soy intake both have been shown to attenuate chronic inflammation. Dietary interventions aiming to attenuate the chronic inflammatory status in dialysis patients need further exploration. PMID- 15876845 TI - Monocytes from dialysis patients exhibit characteristics of senescent cells: does it really mean inflammation? AB - Hemodialysis treatment induces mononuclear cell activation particularly if cellulosic hemodialysis membrane is used. In normal cells, repeated activation induce a process of accelerate cellular senescence. The aim of the present study was to evaluate whether the mononuclear cell activation associated to hemodialysis with cellulosic membranes favors a process of accelerate senescence in mononuclear cells. Our results show that mononuclear cells from patients dialyzed with cellulosic membranes, exhibit: decrease telomere length, increase percentage of cells CD14dim/CD16bright and increase production of IL-1beta, IL1Ra and IL6 cytokines. After culture in vitro, these cells shown increase susceptibility to undergoing spontaneous apoptosis, that is enhanced by IL-4 and prevented by IL-1beta or LPS. All of these characteristics have been reported associated to senescence of monocytes, and not are observed in cells from controls subjects or patients dialyzed with non-cellulosic membranes, suggesting that hemodialysis with cellulosic membranes induce a process of senescence in mononuclear cells. PMID- 15876846 TI - Modifiable risk factors for cardiovascular disease in CKD patients. AB - Risk factors for cardiovascular disease (CVD) have been studied extensively in CKD patients. It can be differentiated between modifiable, potentially-modifiable and non-modifiable risk factors. Nonetheless, even for easily modifiable risk factors there is still a lack of data demonstrating the benefit of common interventions, such as statin treatment for dyslipidemia, improvement of HbA1c levels in diabetic patients, implementation of physical exercise, normalization of Hgb and achievement of adequate dry weight in dialysis patients. This article gives an overview of modifiable and potentially modifiable risk factors and available modification strategies. PMID- 15876847 TI - The cardiovascular burden of the dialysis patient: the impact of dialysis technology. AB - There is widespread recognition that the poor survival rates of dialysis patients, attributed predominantly to cardiovascular disease, need to be addressed and improved. In this paper, we relate diverse aspects of modern dialysis technology with factors that are considered to contribute towards increased mortality and morbidity in the dialysis population. Firstly, we assess the overall cardiovascular burden of the dialysis patient: it is the sum of uraemia-related risk factors (URRF), traditional risk factors and dialysis therapy-related factors. Secondly, we describe how key components of the dialysis procedure may be directly related to the more common URRF: the dialyser and the membrane, microbiological quality of water and dialysate, treatment modality and online monitory equipment. The judicious selection and application of these components may collectively help improve patient outcomes in hemodialysis therapy. PMID- 15876848 TI - Oxidative stress and reactive oxygen species. AB - This article discusses different aspects concerning classification/nomenclature, biochemical properties and pathophysiological roles of reactive oxygen species (ROS) which are pivotal to interpret the concept of oxidative stress. In vitro studies in both the prokaryotes and eukaryotes clearly demonstrate that exogenous or constitutive and inducible endogenous sources of ROS together with cofactors such as transition metals can damage virtually all the biomolecules. This adverse chemistry is at the origin of structural and metabolic defects that ultimately may lead to cell dysfunction and death as underlying mechanisms in tissue degeneration processes. The same biomolecular interpretation of aging has been proposed to embodies an oxidative stress-based process and oxidative stress may virtually accompany all the inflammatory events. As a consequence, ROS have proposed to play several roles in the pathogenesis of chronic-degenerative conditions, such as athero-thrombotic events, neurodegeneration, cancer, some forms of anemia, auto-immune diseases, and the entire comorbidity of uremia and diabetes. Nowadays, the chance to investigate biochemical and toxicological aspects of ROS with advanced biomolecular tools has, if needed, still more emphasized the interest on this area of biomedicine. These technological advancements and the huge information available in literature represent in our time a challenge to further understand the clinical meaning of oxidative stress and to develop specific therapeutic strategies. PMID- 15876849 TI - Calcium, phosphorus and vitamin D disorders in uremia. AB - BACKGROUND: Alterations in calcium, phosphate (P) and vitamin D metabolism play a critical role in the development of secondary hyperparathyroidism (SH), parathyroid hyperplasia and soft tissue and vascular calcification. METHODOLOGY: Studies were performed in uremic dogs and rats fed a low and high P diet over a period of 1-4 months. In addition, in vitro studies were performed in normal parathyroid glands incubated in culture media containing 0.2 mM P (low) or 2.0 mM P (high). RESULTS: Uremic rats maintained on a low P diet did not develop SH or parathyroid hyperplasia. There was an enhancement of p21, the suppressor of the cell cycle, in these parathyroid glands. Opposite results were obtained using a high P diet. There was an enhancement of transforming growth factor-alpha and epidermal growth factor receptor, known enhancers of cell proliferation. In vitro studies demonstrated the direct effect of P on parathyroid hormone secretion. CONCLUSIONS: Early dietary P restriction prevents the development of SH and parathyroid hyperplasia. If dietary P restriction is applied to rats with established SH, there is a significant amelioration of SH and parathyroid hyperplasia. In addition, control of serum P in uremic patients is crucial in the prevention of vascular calcification. PMID- 15876850 TI - Cardiovascular calcification in end stage renal disease. AB - Extensive atherosclerosis and heavy vascular and valvular calcifications are common complications of end stage chronic kidney disease (CKD-V) and are related to a high incidence of cardiovascular events. In CKD-V vascular calcifications occur both in the subintimal space and in the runica media. Intimal calcification is associated with atherosclerosis and is therefore a universal finding. On the contrary, medial calcification is characteristically associated with advanced CKD and diabetes mellitus. Numerous metabolic and endocrine abnormalities, primarily involving calcium and phosphorus metabolism, are found in CKD. Furthermore, CKD-V is believed to be a state of heightened inflammation and oxidative stress. All of these dysfunctions occur early in the course of the renal failure and likely contribute to the development and progression of vascular calcification and atherosclerosis. This review is centred on the pathobiology of vascular calcification in CKD-V, its detection with modern imaging modalities and the therapeutic approaches currently available to slow its progression. PMID- 15876851 TI - The mechanism of calcium deposition in soft tissues. AB - The current understanding of the mechanisms of calcium deposition in soft tissues in chronic kidney disease (CKD) patients has been deeply investigated in the last ten years. Because of higher morbidity and mortality due to cardiovascular disease in dialysis patients compared to general population, several studies showed that extraskeletal calcification may play a major role in the pathogenesis of cardiovascular events in CKD patients. Traditionally, the pathogenesis of vascular and soft tissue calcification has been associated with a passive calcium phosphate deposition. Actually, it is well known that extraskeletal calcification is also related to an active process. In this review, we analyzed some of the factors potentially involved in the pathogenesis of vascular calcification in CKD patients. PMID- 15876852 TI - Biosimilars, generic versions of the first generation of therapeutic proteins: do they exist? AB - This contribution describes the present regulatory status in the EU of biosimilars, the generic versions of the first generation of therapeutic proteins. It points out why and where recombinant protein molecules and low molecular-weight drugs differ in their behaviour and why biosimilars should be handled differently than generic low-molecular-weight drugs. This information is important for practitioners (pharmacists and physicians) while selecting the best supplier of a therapeutic protein. PMID- 15876853 TI - Electrophysiological response to dialysis: the role of dialysate potassium content and profiling. AB - The task of dialysis therapy is, amongst other things, to remove excess potassium (K+) from the body. The need to achieve an adequate K+ removal with the risk of cardiac arrhythmias due to sudden intra-extracellular K+ gradient advises the distribution of the removal throughout the dialysis session instead of just in the first half. The aim of the study was to investigate the electrical behavior of two different K+ removal rates on myocardial cells (risk of arrhythmia and ECG alterations). Constant acetate-free biofiltration (AFB) and profiled K+ (decreasing during the treatment) AFB (AFBK) were used in a patient sample to understand, first of all, the effect on premature ventricular contraction (PVC) and on repolarization indices [QT dispersion (QTd) and principal component analysis (PCA)]. The study was divided into two phases: phase 1 was a pilot study to evaluate K+ kinetics and to test the effect on the electrophysiological response of the two procedures. The second phase was set up as an extended cross over multicenter trial in patient subsets prone to arrhythmias during dialysis. Phase 1: PVC increased during both AFB and AFBK but less in the latter in the middle of dialysis (298 in AFB vs. 200 in AFBK). The PVC/h in a subset of arrhythmic patients was 404 +/- 145 in AFB and 309 +/- 116 in AFBK (p = 0.0028). QT interval (QTc) prolongation was less pronounced in AFBK than in AFB. Phase 2: The PVC again increased in both AFB and AFBK but less in the latter mid-way through dialysis (79 +/- 19 AFB vs. 53 +/- 13 AFBK). Moreover, in the most arrhythmic patients the benefit accruing from the smooth K+ removal rate was more pronounced (103 +/- 19 in AFB vs. 78 +/- 13 in AFBK). CONCLUSION: It is not the K+ dialysis removal alone that can be destabilizing from an electrophysiological standpoint, but rather its removal dynamics. This is all the more evident in patients with arrhythmias who benefit from the K+ profiling during their dialysis treatment. PMID- 15876854 TI - Noninvasive assessment of vascular function. AB - Impaired arterial compliance contributing to increased blood pressure and cardiac workload is well accepted as a major factor in cardiovascular disease. Information on local arterial compliance is obtained when analyzing the deformation of selected arterial segments under stress. A more global measure of arterial compliance is obtained by analyzing the arterial pulse by so-called pulse wave analysis. The arterial pulse, even when measured locally, carries characteristic information from the whole arterial system because of reflection of waves at distinct sites of the arterial system. Pulse wave velocity and the transfer function for pulse transmission is obtained from the combined measurement of arterial pulses at proximal and distal measuring points. Both pulse wave velocity and transfer function importantly, but not exclusively, depend on arterial compliance. The reconstruction of the aortic pulse from peripheral pulse measurements using a population-based transfer function finally provides information on central effects of reduced arterial compliance and increased peripheral resistance which may help in the diagnosis and treatment of vascular disease. PMID- 15876855 TI - New insights in uremic toxicity. AB - The uremic syndrome is characterised by the retention of a host of compounds that in healthy subjects are secreted by the kidneys into normal urine. These compounds disturb many physiologic functions, resulting in toxicity. Many of the responsible compounds remain unknown, however, as well as many patho-physiologic actions of the known retention solutes. In this publication, we review recent new information regarding uremic toxicity. Especially difficult to remove compounds, such as protein bound and larger molecules, seem to play a role. New strategies enhancing their removal might be highly useful. PMID- 15876856 TI - Continuous renal replacement therapy for end-stage renal disease. The wearable artificial kidney (WAK). AB - Daily dialysis offers many benefits but is difficult to implement. CRRT allows dialysis 24/7 but is not suitable for ESRD patients. Thus, the need for a miniaturized ambulatory CRRT device those patients can wear permanently. We report the feasibility, safety and efficiency in uremic pigs, of such a wearable artificial kidney (WAK) that can be worn as a belt, operated with batteries, and weights less than 5 lbs. We used a hollow fiber dialyzer with a surface area of 0.2 sqm. Dialysate was continuously regenerated by a series of cartridges containing several sorbents allowing the use of approximately 375 ml of dialysate. The device includes reservoirs with heparin and electrolytes. Average fluid removal was 100 ml/hr. The Creatinine was 25 ml/min. In 8 hrs the total Creatinine removed was 1 gr, Urea 12 gr, P0.8 gr and K 72 mEq. Weekly st kt/v was extrapolated to approximately 7. There were no side effects. The WAK can be operated safely and continuously 168 hr/week. This would allow for all the advantages of daily dialysis and reduce morbidity and mortality in the ESRD population. It will also reduce cost and manpower utilization. PMID- 15876857 TI - Slow continuous intravenous plasmapheresis (SCIP): clinical applications and hemostability of extracorporeal ultrafiltration. AB - An intravenous plasmapheresis catheter which excludes >99.4% of platelets from external ultrafiltration circuits is currently undergoing safety and efficacy trials for fluid removal from NYHA class II-IV congestive heart failure patients resistant to diuretic drug therapy. In animals, the SCIP catheter allowed a four fold increase in ultrafiltration efficiency without hemolysis, hemoinstability or external cartridge changes in 72 hours of treatment. Further, systemic anticoagulation was not required. These techniques might be envisioned for treatment of fluid overload in heart failure, surgery or trauma and may have applications in therapeutic apheresis, venous thrombosis, liver disease or autologous tissue engineering. PMID- 15876858 TI - Membraneless dialysis--is it possible? AB - Direct contact between uremic blood and a fluid capable of receiving uremic toxins is possible. Such contact by itself is, however, not beneficial because the selection of molecules that are removed is dependent on diffusion coefficients in blood. This selection is inadequate and would result in the exhaustion of a patient's albumin pool before useful reduction in the urea pool was achieved. Direct contact that is accomplished by sandwiching blood between two layers of a sheathing fluid, followed by diafiltration of the sheathing fluid through conventional membranes and recirculation of the sheathing fluid, is possible. This adaptation of membraneless transport of molecules from blood eliminates almost all contact of blood with solid artificial surfaces and the subsequent diafiltration and recirculation of the sheathing fluid allows precise control of what is removed from the system. Slightly hyperosmotic protein is carried back by the recirculating sheathing fluid. Only solutes and water that pass the diafilter, which operates on a cell-free fluid, are able to leave the system. The system depends strongly on the ability to keep cells out of the sheathing fluid. Preliminary results and earlier reports indicate that this separation is possible and more precise measurements are underway. A quantitative design of a wearable dialyzer based on a circulating sheathing fluid is presented. PMID- 15876859 TI - Inhibition of P-glycoprotein-mediated multidrug resistance by unfractionated heparin: a new potential chemosensitizer for cancer therapy. AB - Anticoagulant treatment with heparins is frequently used to prevent venous thromboembolism in cancer patients. In the present study, we investigated the ability of unfractionated heparin (UFH) to inhibit P-glycoprotein (Pgp)-mediated multidrug resistance (MDR) on human breast cancer cell line (MDA-MB231) and its doxo-resistant subline. Results were a compared to the classic reversing agent, Verapamil (Ver), used, as reference at 50 microM concentration. We analysed the Pgp function by calcein acetoxymethylester (calcein-AM) uptake, a fluorescent marker substrate, before and after in vitro exposure to UFH at clinically achievable dose of 20 U/ml. The mean percentage of calcein-AM retained into cancer cells after 3 and 12 h were 32 +/- 10.9 and 45 +/- 12.3, respectively, for UFH pretreated cells and 25.3 +/- 8.7 and 29.4 +/- 10.4, respectively, for Ver pretreated cells when compared to control cells, receiving only medium. Pgp activity was studied by measuring intracellular drug accumulation in doxo resistant subline, treated (2 h) with either UFH or Ver, prior exposure (2 h) at different doxo concentrations (2, 4 and 8 microM). The mean percentage of remaining intracellular doxo were 55.4 +/- 4.5 , 51.4 +/- 3.9 and 50 +/- 1.8 percent, respectively for UFH treated cells, and 44.1 +/- 5.8, 39.3 +/- 4.4 and 19.4 +/- 8.6%, respectively, for Ver treated cells as compared with control cells, receiving only doxo. These results were consistent with the increase of sensitivity to doxo of the same doxo-resistant subline resulting in a 2.2, 2.6 and 2.2-fold increase, respectively, for UFH-doxo combination and 2.2, 2.5 and 2.0-fold respectively, for Ver-doxo combination respect to cells receiving doxo alone, as assessed by MTT test. In conclusion, these findings demonstrate the potentiating effect in vitro of UFH on doxo accumulation and cytotoxicity in the MDA-231 cell line and its doxo-resistant subline and suggest that UFH could to be used, as an potential chemosensitizer, in clinical chemotherapy for increasing in vivo, the efficacy of the anticancer treatment. PMID- 15876860 TI - Localization of phosphorylated forms of Bcl-2 in mitosis: co-localization with Ki 67 and nucleolin in nuclear structures and on mitotic chromosomes. AB - Bcl-2 phosphorylation is a normal physiological process occurring at mitosis or during mitotic arrest induced by microtubule damaging agents. The consequences of Bcl-2 phosphorylation on its function are still controversial. To better understand the role of Bcl-2 phosphorylation in mitosis, we studied the subcellular localization of phosphorylated forms of Bcl-2. Immunofluorescence experiments performed in synchronized HeLa cells indicate for the first time that mitotic phosphorylated forms of Bcl-2 can be detected in nuclear structures in prophase cells together with nucleolin and Ki-67. In later mitotic stages, as previously described, phosphorylated forms of Bcl-2 are localized on mitotic chromosomes. In addition, we demonstrate that Bcl-2 in these structures is at least in part phosphorylated on the T56 residue. Then, coimmunoprecipitation experiments reveal that, in cells synchronized at the onset of mitosis, Bcl-2 is present in a complex with nucleolin, cdc2 kinase and PP1 phosphatase. Taken together, these data further support the idea that Bcl-2 could have a new function at mitosis. PMID- 15876861 TI - Gam1 and the SUMO pathway. AB - Post-translational modifications of proteins have critical roles in many cellular processes because they can cause rapid changes in the functions of preexisting proteins, multiprotein complexes and subcellular structures. Sumoylation, a ubiquitin-like dynamic and reversible post-translational modification system, is an enzymatic cascade leading to the covalent attachment of SUMO to it target proteins. This modification involves three steps and different enzymes: SUMO activating enzyme E1 (SAE1/SAE2), SUMO-conjugating enzyme E2 (UBC9), SUMO ligases E3s, and SUMO cleaving enzymes. Although the identification of SUMO-modified substrates has progressed rapidly, the biological function of SUMO and regulation of SUMO conjugation are still not well understood. Some viral proteins have been identified as substrates for SUMO modification as well as altering the sumoylation status of host cell proteins. We have been studying an unusual adenoviral protein, Gam1, a strong and global transcriptional activator of both viral and cellular genes that inactivates HDAC1. We have recently expanded the known functions of Gam1 by demonstrating that Gam1 also inhibits the SUMO pathway by interfering with the activity of E1 heterodimer (SAE1/SAE2), leading to the accumulation of SUMO-unmodified substrates. Our data provides a clear example of the effects of a viral infection on host sumoylation and supports the idea that viruses have multifunctional protein that can target essential biochemical pathways. PMID- 15876862 TI - MTAP homozygous deletion: an Achilles heel of human cancers ready for clinical use? PMID- 15876863 TI - A novel DNA vaccine containing four mimicry epitopes for gastric cancer. AB - Gastric cancer is one of the most common malignant tumors in China. This paper focuses on the development of a DNA vaccine containing four mimotopes of MG7Ag for gastric cancer (multi-epitope vaccine). By inoculating BALB/c mice, the vaccine was characterized and compared with a similar vaccine containing only one mimotope (mono-epitope vaccine) and other controls. Cellular ELISA indicated that serum titer of antibody against MG7Ag was significantly higher in mice immunized with the multi-epitope vaccine than that in the group immunized with the mono epitope vaccine (0.8627 vs 0.6754, P < 0.05). And ELISPOT assay showed that the number of INF-gamma spots induced by multi-epitope vaccine was significantly larger than that of the group immunized with mono-epitope vaccine(93.3 vs 70.7, P < 0.05). Two weeks after tumor challenge, the weight of tumor in each mouse was evaluated, and the tumor masses formed in the mice immunized with multi-epitope vaccine were markedly smaller than those formed in the mice immunized with mono epitope vaccine. These studies demonstrated that both humoral and cellular response were induced by the two vaccines and the efficiency of multi-epitope vaccine is stronger than that of the mono-epitope vaccine. PMID- 15876864 TI - MdmX inhibits ARF mediated Mdm2 sumoylation. AB - Mdm2, by virtue of an intrinsic E3 ubiquitin ligase activity, is capable of autoubiquitination and the ubiquitination of the p53 tumor suppressor protein. Additionally, Hdm2 has been reported to undergo a p14ARF-dependent sumoylation with concurrent Hdm2 stabilization. In this present work, we report that MdmX can undergo ARF-mediated sumoylation similar to that reported for Mdm2. When coexpressed, MdmX overexpression results in a dose-dependent inhibition of Mdm2 sumoylation and a concurrent increase in Mdm2 ubiquitination. This switch from Mdm2 sumoylation to Mdm2 ubiquitination may explain the destablization of Mdm2 previously observed in cells overexpressing both ARF and MdmX. Given that MdmX can heterodimerize with Mdm2 and separately associate with ARF we employed a series of MdmX mutants to examine how MdmX blocks Mdm2 sumoylation. A MdmX miniprotein capable of binding to ARF, but not p53 or Mdm2 was able to competitively inhibit Mdm2 sumoylation and reverse ARF mediated activation of p53 transactivation. Taken together, these results demonstrate that MdmX can affect post-translational modification and stability of Mdm2 and p53 activity through interaction with ARF. PMID- 15876865 TI - The anaphase promoting complex: a critical target for viral proteins and anti cancer drugs. AB - The study of animal viruses has provided extraordinary insights into cell cycle dynamics and tumor biology. The significance of the p53 and Rb tumor suppressor proteins, for example, was discovered due to their interactions with viral oncogenes. In the past several years, investigations with four viral proteins, human immunodeficiency virus type 1 (HIV-1) vpr, adenovirus E4orf4, chicken anemia virus (CAV) apoptin and human T lymphotropic virus type I (HTLV-I) Tax, have indicated that there are also critical viral targets involved in G2/M control. In particular, recent studies with E4orf4 and apoptin have shown that they induce G2/M arrest by targeting and inhibiting the anaphase-promoting complex/cyclosome (APC/C). Notably, these two viral proteins induce apoptosis selectively in transformed cells in a p53-independent manner; thus pathways affected by these proteins are of significant therapeutic interest. Further investigation of the underlying mechanism of G2/M arrest and subsequent apoptosis induced by viral APC/C inhibitors may shed light on the mechanisms of current cancer therapies and provide the foundation for developing novel therapeutic targets. PMID- 15876866 TI - Evidence that DNA damage detection machinery participates in DNA repair. AB - The toroidal Rad9-Rad1-Hus1 checkpoint complex (9-1-1) is structurally similar to the proliferating cell nuclear antigen (PCNA), which serves as a sliding clamp platform for DNA replication and repair. 9-1-1 has been characterized as a sensor of DNA damage that functions in concert with the checkpoint control proteins ATM and ATR. However, recent data suggest that the 9-1-1 complex and its individual Rad9 component serve different and multiple functions in cells by sensing DNA damage, stimulating apoptosis, and regulating gene transcription. Recently it was reported that 9-1-1 interacts with and/or stimulates components of the base excision repair (BER) pathway including the S. pombe MutY homolog (MYH), human polymerase beta (Polbeta), and flap endonuclease 1 (FEN1). Furthermore, preliminary results indicate a stimulation of DNA ligase I. In this review, the likely direct participation of 9-1-1 in DNA repair is discussed. PMID- 15876867 TI - Cyclic peptides, a chemical genetics tool for biologists. AB - The genomics era has presented scientists with an overwhelming amount of sequence information that encodes for a continuously expanding array of metabolic pathways and regulatory networks. In order to unravel this elaborate maze, effective approaches need to be pioneered, and cyclic peptides represent an exciting new tool that can aid this challenging task. There are considerable benefits to cyclic peptides including enhanced protein binding affinity and metabolic stability, but perhaps most important is the ability to biologically synthesize them using intein chemistry. This capacity for genetic encoding allows biologists access to diverse, intracellular small-molecule libraries that can be tailored for numerous chemical genetics approaches. An increasing number of molecular strategies are being developed to aid in the implementation of cyclic peptides in forward and reverse genetics experiments, which will provide researchers with a powerful toolbox to complement existing genomic methodologies. PMID- 15876868 TI - Genetic response to DNA damage: proapoptotic targets of RhoB include modules for p53 response and susceptibility to Alzheimer's disease. AB - Knockout mouse studies indicate that the small GTPase RhoB is critical for apoptosis triggered by genotoxic stress in transformed mouse cells. However, the mechanisms used by RhoB to sensitize cells to cell death are obscure. To gain insight into this question, we compared the genetic response of cells with different rhoB genotypes to the DNA damaging anticancer drug doxorubicin (DOX). The microarray hybridization strategy focused on events occurring by 6 hr of DOX treatment, preceding the execution phase of RhoB-dependent apoptosis by 12-16 hr. Genes controlling cytoskeletal organization, adhesion, transcription, trafficking, apoptosis, and protein turnover were represented prominently. Gene clustering revealed a module of p53 target genes, suggesting that RhoB may modify the p53 response, and a module for susceptibility to Alzheimer's disease, suggesting a link between RhoB and age-associated dementia. The findings of this study suggest mechanisms by which RhoB may act to elevate the sensitivity of cells to apoptosis following genotoxic stress. PMID- 15876869 TI - Mutated PI 3-kinases: cancer targets on a silver platter. AB - The PI3K signaling pathway is upregulated in numerous cancers. The catalytic subunit p110alpha of PI3K shows hot spot mutations in nearly 30% of several types of solid tumors. The most prominent of these mutations result in gain of enzymatic function, activate Akt signaling and induce oncogenic cellular transformation. The mutated p110alpha proteins are ideal targets for specific small molecule inhibitors that discriminate between the oncogenic and the wild type forms of the enzyme. Such inhibitors could become highly effective anti cancer drugs. PMID- 15876870 TI - Functional genomic analysis of cell division by endoribonuclease-prepared siRNAs. AB - The emergence of RNA interference (RNAi) technology has revolutionized functional genomic analyses in cell biology, including the study of cell division. In particular, the introduction of short interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) has facilitated loss-of-function studies in mammalian cell lines. We have pioneered the rapid and cost-efficient generation of libraries of endoribonuclease-prepared short interfering RNAs (esiRNAs) for large-scale genetic screens in mammalian tissue culture cells. Our first pilot screen of 5305 genes in human HeLa cells identified novel genes required for cell division, establishing esiRNA as a sophisticated method for loss-of-function screens. Future genome-wide studies with esiRNA, using image-based assays and video microscopy, will help provide novel insights on the molecular mechanisms that govern cell division in mammalian cells. PMID- 15876871 TI - CAK-Cyclin-dependent Activating Kinase: a key kinase in cell cycle control and a target for drugs? AB - The Cyclin-dependent kinase (CDK) Activating Kinase (CAK) is responsible for the activating phosphorylation of CDK1, CDK2, CDK4 and CDK6 and regulation of the cell cycle. The kinase is composed of three subunits: CDK7, Cyclin H and MAT1 (menage a trois). Together with six other subunits, CAK is also part of the general transcription factor TFIIH where it is involved in promoter clearance and progression of transcription from the preinitiation to the initiation stage. CAK is required for cell cycle progression, which suggests that CDK7 could be a target for cancer therapy. However its role in transcription and its ubiquitous presence raise sensible concerns about possible toxicity of its inhibitors. The recently determined structure of CDK7 allows the design of inhibitors with differential specificity for the different CDKs. We review the role of CAK in different biological processes and evaluate the biological evidence for CDK7 as a possible pharmacological target. PMID- 15876872 TI - Inhibition of caspase 9 and not caspase 8 mediated apoptosis may determine clinical response to chemotherapy in primary nodal diffuse large B-cell lymphomas. AB - Clinical response to chemotherapy in patients with diffuse large B-cell lymphomas is poorly predictable. We demonstrated that an expression profile consistent with inhibition of the stress-induced, caspase 9 mediated apoptosis pathway predicts poor clinical response to chemotherapy, whereas an expression profile consistent with inhibition of only the death-receptor induced, caspase 8 mediated pathway was associated with an excellent response to chemotherapy and favorable outcome. Furthermore, functional analysis of apoptosis sensitivity showed that lymphomas with a caspase 9 inhibition profile were indeed relatively resistant to Etoposide induced apoptosis. Determining the functional status of both apoptosis signaling pathways may accurately identify patients at high risk for fatal outcome and may have implications for alternative therapies triggering the death receptor mediated apoptosis pathway. PMID- 15876873 TI - Stromal responses to carcinomas of the pancreas: juxtatumoral gene expression conforms to the infiltrating pattern and not the biologic subtype. AB - If there is a "science" of tumor-stromal interactions, there must be a set of biologic rules that are organ-site dependent. One way to explore this hypothesis would be to compare the patterns of gene expression of two biologically distinct neoplasms that arise within the same organ site. Using nonradioactive in situ hybridization, we evaluated the gene expression patterns of three genes previously shown to be robust markers of the juxtatumoral stroma within eight infiltrating ductal adenocarcinoma of the pancreas (ApoC1, ApoD and MMP11), and compared these patterns to those associated with seven infiltrating colloid and tubular carcinomas arising in association with intraductal papillary mucinous neoplasms (IPMNs), a histologically distinct form of primary carcinoma of the pancreas, two surgically resected samples of chronic pancreatitis and two surgically resected pancreatic cancer liver metastases. Robust juxtatumoral stromal expression was noted for all three markers within all eight conventional infiltrating ductal adenocarcinoma tissues, but not in samples of chronic pancreatitis. Among the carcinomas arising within an IPMN, expression for all three markers was also noted for five of seven infiltrating carcinomas analyzed. However, when labeling for these three markers was analyzed with respect to infiltrative growth pattern, positive labeling was only seen in areas of tubular (ductal-type) growth and not in areas of colloid carcinoma. This observation was further supported by two infiltrating carcinomas arising in an IPMN that showed both tubular and colloid growth patterns within the same neoplasm indicating the host stromal response observed may relate to infiltrative growth pattern rather than the biology of the primary tumor type. Moreover, these robust patterns within conventional infiltrating ductal adenocarcinomas were not retained within matched metastases to the liver, indicating the importance of the tumor microenvironment in the host stromal response. Juxtatumoral stroma was found to be composed of a least two cell types, tumor-infiltrating macrophages and fibroblasts, highlighting the complexity of tumor-stromal interactions within an infiltrating carcinoma. Since the juxtatumoral gene expression response is the strongest indication of direct communication between stroma and cancer cells, we provide evidence of a stereotypical response to infiltrative growth that might predominate in tumor-stromal interactions independent of cancer type, a finding with clinical implications for therapeutic modalities that target this response in human tumors. PMID- 15876874 TI - p14ARF interacts with the SUMO-conjugating enzyme Ubc9 and promotes the sumoylation of its binding partners. AB - The p14ARF tumour suppressor regulates a series of cell cycle regulatory proteins to promote cell cycle arrest in response to abnormal hyperproliferative growth stimuli. p14ARF alterations are common in human cancers and, when inherited, confer susceptibility to cutaneous melanoma. We now propose that the mechanism of p14ARF action may involve the covalent modification of its binding partners with the small ubiquitin-related protein SUMO-1. In particular, we demonstrate that p14ARF interacts with the SUMO E2 conjugating enzyme, Ubc9 and enhances the sumoylation of its binding partners, hdm2, E2F-1, HIF-1alpha, TBP-1 and p120E4F. Furthermore, p14ARF-induced sumoylation is abrogated by a subset of melanoma associated p14ARF mutations. These results provide a mechanism for p14ARF action through a common modification of diverse binding partners. PMID- 15876875 TI - The regulation of S phase initiation by p27Kip1 in NIH3T3 cells. AB - Most eukaryotic cells become committed to divide once DNA synthesis begins. This critical event is controlled in yeast and Xenopus oocytes by the degradation of cyclin inhibitory proteins, while in mammalian cells over-expression of cyclin E or cyclin D1 promotes rapid entry into DNA synthesis. Curiously, however, this over-expression also lengthens S phase and promotes DNA damage. To directly assess the roles of the cyclin inhibitory protein p27Kip1 (p27) and of cyclin D1 in the regulation of DNA synthesis initiation in mammalian cells, we have utilized a quantitative cytometric approach for the study of cell cycle control in actively proliferating cultures. As evidence that p27 plays a direct role in regulating entry into S phase, we find that its levels fall at the time of DNA synthesis initiation in NIH3T3 cells, and that its suppression shortens G1 phase and shortens the length of the entire cell cycle. In this function, however, the action of p27 appears to be linked with cyclin D1. G1/S phase transition is efficiently blocked by p27 unless excess cyclin D1 is present. These two proteins are coordinately regulated by the cell, and are maintained at a nearly constant ratio throughout an actively proliferating culture. We propose that p27 directly regulates the initiation of DNA synthesis in NIH3T3 cells, and that cyclin D1 serves to modulate this activity. PMID- 15876876 TI - The Plk1 Polo box domain mediates a cell cycle and DNA damage regulated interaction with Chk2. AB - Polo-like kinase 1 (Plk1) regulates multiple processes during mitosis. Chk2 is a tumor suppressor that participates in DNA damage checkpoint signaling cascades. Plk1 phosphorylates, colocalizes with, and interacts with Chk2, suggesting interconnection of DNA damage checkpoints and mitotic regulation. However, the function of their association is unknown. Here, we show that the interaction between Chk2 and Plk1 is cell cycle-regulated, with a peak in mitosis. DNA damage in G2 and M phases but not in S phase induces dissociation of Plk1 and Chk2. In vitro, the Plk1 PBD binds phosphorylated Chk2, and mediates an interaction independent of other eukaryotic proteins. Additionally, a phosphopeptide encompassing phosphoT68 of Chk2 binds Plk1 in a PBD-dependent manner, and stimulates Plk1 activity. These results identify potential mechanisms for interaction and inter-regulation of these two protein kinases. PMID- 15876877 TI - Temporal control of cell cycle gene expression mediated by E2F transcription factors. AB - Various studies now point to the role of cooperative transcription factor interactions as a mechanism to achieve the necessary specificity in promoter recognition. An example can be seen in the action of E2F proteins where interactions with other transcription factors not only provides this general specificity but also the specificity that distinguishes function of the individual E2F proteins. This includes both the activation and repression functions of E2Fs and has now also been extended to the control of G2/M regulated genes in addition to the G1/S E2F targets. These studies further highlight events linking the control of G1/S genes with G2/M, providing a mechanism to achieve the temporal control of gene expression as cells move through the cell cycle. PMID- 15876879 TI - Outcomes of an infection prevention project focusing on hand hygiene and isolation practices. AB - Nosocomial infections are a major health problem for hospitalized patients and their families. Since the 1800s, hand hygiene has been recognized as the single best method to prevent the spread of pathogens and nosocomial infections. Despite this fact, many healthcare workers do not adhere to hand hygiene policies. The Centers for Disease Control and Prevention issued a guideline for hand hygiene practices in 2002. Multifaceted approaches to improve hand hygiene have been shown to increase compliance among healthcare workers and subsequently reduce infections. A performance improvement project was initiated to implement this guideline and other strategies to prevent nosocomial infection. This article summarizes the performance improvement processes and the preliminary outcomes on adherence to infection prevention policies related to hand hygiene and isolation practices. Clinically and statistically significant increases were noted for hand hygiene prior to patient care and in wearing masks when indicated. Nurses and patient care technicians had the greatest increases in compliance. Increases in hand hygiene after patient contact and wearing of gown and gloves were also noted, but results were not statistically significant. Nosocomial infection rates from antibiotic-resistant organisms decreased in the first surveillance, but rates increased during the 1-year surveillance. Consumption of alcohol-based foam disinfectant doubled from baseline. Findings are consistent with other published studies. The project will continue with further reinforcement and education over the second year. PMID- 15876880 TI - Quality improvement in backrest elevation: improving outcomes in critical care. AB - The positioning of critically ill patients is an independent nursing decision, often has multiple rationales, and may significantly affect morbidity and mortality. Recent evidence suggests that backrest elevation in critically ill patients may reduce ventilator-associated pneumonia. However, use of recommended levels of backrest elevation is infrequent in the critical care environment. In addition, published guidelines for backrest elevation to reduce pneumonia conflict with those for protecting skin integrity. This article reviews the benefits and complications of backrest elevation, data related to current positioning practices, and recommendations for backrest elevation. A quality improvement process to guide evidence-based care related to backrest positioning is also described. PMID- 15876881 TI - The use of common continuous monitoring parameters: a quality indicator for critically ill patients with sepsis. AB - Sepsis is a common source of morbidity and mortality among critically ill patients, and targeting measures to promote early recognition and treatment of sepsis is at the forefront of many critical care initiatives. Starting formally in 1992, with the publication of the definitions of sepsis, continuous monitoring of several common physiologic parameters, including electrocardiogram, blood pressure, and oxygen saturation, have been advocated as important in the early identification and treatment of patients with sepsis. The descriptive study detailed in this article was conducted to assess the perceptions and clinical continuous physiologic monitoring practices of experienced critical care clinicians with regard to their use of common physiologic monitoring parameters in the care of patients with sepsis. A convenience sample of 100 physicians and 517 nurses completed a 20-item survey assessing perceptions and clinical monitoring practices related to the care of patients with sepsis. Results indicated that the basic parameters of electrocardiogram, invasive blood pressure, pulmonary arterial catheter monitoring, and oxygen saturation all have value in the recognition and treatment of patients with sepsis. The majority of clinicians used these parameters routinely and felt they were necessary for patient care. These results indicate that clinical practice is in concordance with current practice recommendations. PMID- 15876882 TI - Cardiovascular surgeon and acute care nurse practitioner: collaboration on postoperative outcomes. AB - Changes occurring in the healthcare environment require healthcare delivery systems to provide high quality care services with increased efficiency and cost effectiveness. Healthcare systems are encouraged to use less expensive care providers for medical management responsibilities while maintaining or increasing quality of patient care. Accompanying the changes in healthcare delivery modes is the parallel rise in patient acuity levels related to chronic illnesses of patients admitted for cardiac services such as cardiovascular surgeries. This retrospective, 2-group comparison study examined patient and economic outcomes between 2 groups of adult patients for whom postoperative cardiovascular care was directed by either cardiovascular surgeons alone or cardiovascular surgeons in collaboration with acute care nurse practitioners. Outcome measures included length of stay and cost for an episode of care. Findings revealed that when cardiovascular surgeons, in collaboration with acute care nurse practitioners, directed postoperative care, the length of stay decreased 1. 91 days and total cost decreased 5,038.91 dollars per patient. PMID- 15876883 TI - "It's the people that make the environment good or bad": the patient's experience of the acute care hospital environment. AB - A review of contemporary nursing research reveals a tendency to focus on select aspects of the hospital environment such as noise, light, and music. Although studies such as these shed light on discrete aspects of the hospital environment, this body of literature contributes little to an understanding of the entirety of that world as the patient in the sickbed experiences it. The purpose of the study detailed in this article was to describe the patient's experience of the acute care hospital environment. Nondirective, in-depth phenomenological interviews were conducted, then transcribed verbatim, and analyzed for themes. Against the backdrop of "I lived and that's all that matters," there were 3 predominant themes in patients' experience of the acute care environment: (1) disconnection/connection, (2) fear/less fear, and (3) confinement/freedom. In this environment, human-to-human contact increased security and power in an environment that was described as sterile, disorienting, and untrustworthy. Acute and critical care nurses and other caregivers can use the findings to create less noxious hospital environments. PMID- 15876885 TI - The changing face of critical care medicine: nurse practitioners in the pediatric intensive care unit. AB - Over the last 50 years, healthcare has undergone countless changes. Some of the important changes in recent years have been budget cuts, decreased resident work hours, and increased patient acuity. The need for additional clinical expertise at the bedside has resulted in nurse practitioners becoming an integral part of the healthcare delivery team. To date, little has been published regarding the role of the nurse practitioners in intensive care units. This article outlines how one pediatric hospital has successfully utilized nurse practitioners in the intensive care unit. PMID- 15876886 TI - Munchausen syndrome by proxy: a case report. AB - Munchausen syndrome by proxy is difficult to diagnose unless healthcare providers are astute to its clinical features and management. A case is presented to educate nurses and advanced practice nurses, of the nursing, medical, legal, and social complexities associated with Munchausen syndrome by proxy. This article also provides a brief review of the definition of Munchausen syndrome by proxy, its epidemiology, common features of the perpetrator, implications for healthcare personnel, and the legal and international ramifications of Munchausen syndrome by proxy. PMID- 15876887 TI - Pediatric catheter-related bloodstream infections: latest strategies to decrease risk. AB - Central venous catheters are often mandatory devices when caring for critically ill children. They are required to deliver medications, nutrition, and blood products, as well as for monitoring hemodynamic status and drawing laboratory samples. Any foreign object that is introduced to the body is at risk for infection. Central venous catheters carry a particularly high risk of infection and these infections can be life threatening. Advanced practice nurses possess the power to influence catheter-related line infections in their critical care units. Understanding current recommendations for catheter material selection, site selection, site preparation, and site care can affect rates of catheter related bloodstream infections. This article discusses risk factors for developing catheter-related bloodstream infections in critically ill children, as well as measures to decrease incidence of catheter-related bloodstream infections, including a review of recommendations from the Centers for Disease Control and Prevention. PMID- 15876888 TI - Hyperosmolar therapy in the treatment of severe head injury in children: mannitol and hypertonic saline. AB - Traumatic brain injury is the result of a primary, acute injury and is complicated by the development of secondary injury due to hypotension and hypoxia. Cerebral edema due to brain injury compromises the delivery of essential nutrients and alters normal intracranial pressure. The Monroe-Kellie Doctrine defines the principles of intracranial pressure homeostasis. Treatment for intracranial hypertension is aimed at reducing the volume of 1 of the 3 intracranial compartments, brain tissue, blood, and cerebrospinal fluid. Hyperosmolar therapy is one treatment intervention in the care of patients with severe head injury resulting in cerebral edema and intracranial hypertension. The effect of hyperosmolar solutions on brain tissue was first studied nearly 90 years ago. Since that time, mannitol has become the most widely used hyperosmolar solution to treat elevated intracranial pressure. Increasingly, hypertonic saline solutions are being used as an adjunct to mannitol in basic science research and clinical studies. Hyperosmolar solutions are effective in reducing elevated intracranial pressure through 2 distinct mechanisms: plasma expansion with a resultant decrease in blood hematocrit, reduced blood viscosity, and decreased cerebral blood volume; and the creation of an osmotic gradient that draws cerebral edema fluid from brain tissue into the circulation. The pediatric section of the Society of Critical Care Medicine and the World Federation of Pediatric Intensive and Critical Care Societies adapted previously published guidelines for the treatment of adult brain injury into guidelines for the treatment of children with traumatic brain injury. These guidelines offer recommendations for the management of children with severe head injury, including the use of mannitol and hypertonic saline to treat intracranial hypertension. Acute and critical care pediatric advanced practice nurses caring for children with severe head injury should be familiar with management guidelines and the use of hyperosmolar solutions. The purpose of this article is to assist the advanced practice nurse in understanding the role of hyperosmolar therapy in the treatment of pediatric traumatic brain injury and review current guidelines for the use of mannitol and hypertonic saline. PMID- 15876889 TI - Management of increased intracranial pressure in the critically ill child with an acute neurological injury. AB - Increased intracranial pressure reflects the presence of mass effect in the brain and is associated with a poor outcome in children with acute neurological injury. If sustained, it has a negative effect on cerebral blood flow and cerebral perfusion pressure, can cause direct compression of vital cerebral structures, and can lead to herniation. The management of the patient with increased intracranial pressure involves the maintenance of an adequate cerebral perfusion pressure, prevention of intracranial hypertension, and optimization of oxygen delivery. This article reviews the neurological assessment, pathophysiology, and management of increased intracranial pressure in the critically ill child who has sustained an acute neurological injury. PMID- 15876890 TI - Oncological emergencies in the pediatric intensive care unit. AB - The overall 5-year survival rate of children with cancer has now reached 77%, an increase of about 45% in the past 25 years. Newer therapies, including hematopoietic cell transplantation and cutting edge chemotherapeutics evolving in the form of molecular and biological cell targeted agents, are being researched and developed and are responsible for the change in survival rates over time. Also, despite the national trend toward hospice and palliative care, children with chronic and life threatening illnesses, continue to die in the hospital setting, often in the intensive care unit. Previous studies of children with complications of cancer and its therapy document poor outcomes among those who do require intensive care. These trends are changing, however, currently leaving a hopeful, optimistic view of the outcome in children with cancer complications admitted to the pediatric intensive care unit. It is imperative that nurses and intensive care staff understand pediatric cancer and its potential emergent consequences in order to respond to the symptoms of life threatening events. PMID- 15876891 TI - The case against renal dose dopamine in the pediatric intensive care unit. AB - The use of dopamine for the treatment of renal insufficiency has become a controversial issue. Dopamine exerts its effects on the kidneys through activity on the catecholamine receptors and by its diuretic and natriuretic properties. Utilization of renal dose dopamine to increase renal blood flow has been considered beneficial for preservation of renal function for over 30 years. The hypothesis proposed was that increasing urine volume must indicate improving renal function, particularly in oliguric patients. However, recent clinical trials in adult and pediatric patients have not only failed to demonstrate any benefit, but have also suggested that this therapy may actually have detrimental effects. This article reviews basic pharmacology and physiologic effects and the potential adverse effects of "renal dose dopamine." It also examines the results of clinical trials, in both pediatric and adult patients, that evaluated its usefulness for the treatment of renal insufficiency. PMID- 15876892 TI - Atrial septal defect treatment options. AB - Atrial septal defects have traditionally been repaired by surgical closure. Recently, transcatheter device closure has increasingly been used with excellent results. Although there is limited comparative research evaluating long-term outcomes of the transcatheter technique, preliminary data reveal significantly fewer complications and shorter hospital stays than those reported for surgical repair. This article reviews relevant literature comparing efficacy, cost, and complications of the transcatheter device procedures with the surgical closure of ASDs. PMID- 15876896 TI - The challenge of teaching psychopharmacology and improving clinical practice. PMID- 15876897 TI - QTc variability in schizophrenia patients treated with antipsychotics and healthy controls. AB - QTc prolongation is associated with the administration of some antipsychotics but the QTc interval is also known to vary physiologically. There is little published evidence about changes in QTc variability during treatment with antipsychotics. In this prospective investigation, we analyzed ECGs in 61 patients suffering from a schizophrenic disorder who were treated with different antipsychotics and 31 sex- and age-matched healthy controls. We found no differences in QTc intervals nor in QTc variability between patients and controls. Our results raise the question of the clinical relevance of a single ECG for diagnostics of cardiac complications in schizophrenia patients and suggest the need to conduct ECG monitoring in patients at high risk for cardiac complications during antipsychotic treatment. PMID- 15876898 TI - Influence of topiramate on olanzapine-related adiposity in women: a random, double-blind, placebo-controlled study. AB - The aim of this study was to compare the efficacy of topiramate versus a placebo in the treatment of adiposity in women undergoing olanzapine therapy. We also assessed changes health-related quality of life, the patient's actual state of health, and psychologic impairments. The 10-week, random, double-blind, placebo controlled study included 43 women who had been treated with olanzapine (mean dose 7.8 +/- 3.6 in the topiramate group and 7.2 +/- 3.1 in the placebo group) and had gained weight as a side effect. The subjects were randomly assigned to topiramate (n = 25) or a placebo (n = 18). Primary outcome measures were weight checks and self-reported changes on the scales of the SF-36 Health Survey, Bf-S Scale of Well-Being, and the Adjective Checklist EWL-60-S. Weight loss was observed and was significantly more pronounced in the topiramate-treated group (difference in weight loss between the 2 groups: 5.6 kg, 95% CI = -8.5, -3.0, P < 0.001). In comparison with the placebo group, significant changes on 7 (7/8) scales of SF-36 Health Survey (all P < 0.001), on all 6 scales of the EWL-60-S, and on the Bf-S were observed in the topiramate-treated subjects after 10 weeks. All patients tolerated topiramate well. Topiramate appears to be a safe and effective agent in the treatment of weight gain that occurred during olanzapine treatment. Significantly positive changes in health-related quality of life, the patient's actual state of health, and psychologic impairments were observed. PMID- 15876899 TI - A double-blind placebo-controlled trial of bupropion sustained-release for smoking cessation in schizophrenia. AB - The objective of this study was to examine the efficacy of bupropion for smoking cessation in patients with schizophrenia. Adults with schizophrenia who smoked more than 10 cigarettes per day and wished to try to quit smoking were recruited from community mental health centers, enrolled in a 12-week group cognitive behavioral therapy intervention, and randomly assigned to receive either bupropion sustained-release 300 mg/d or identical placebo. Fifty-three adults, 25 on bupropion and 28 on placebo, were randomized, completed at least 1 postbaseline assessment and were included in the analysis. The primary outcome measures were 7-day point prevalence abstinence in the week after the quit date (week 4) and at the end of the intervention (week 12). Subjects in the bupropion group were significantly more likely to be abstinent for the week after the quit date (36% [9/25] vs. 7% [2/28], P = 0.016) and at end of the intervention (16% [4/25] vs. 0%, P = 0.043). Subjects in the bupropion group also had a higher rate of 4-week continuous abstinence (weeks 8-12) (16% [4/25] vs. 0%, P = 0.043) and a longer duration of abstinence (4.2 [3.2] weeks vs. 1.8 [0.96] weeks, t = 2.30, P = 0.037). The effect of bupropion did not persist after discontinuation of treatment. Subjects in the bupropion group had no worsening of clinical symptoms and had a trend toward improvement in depressive and negative symptoms. We conclude that bupropion does not worsen clinical symptoms of schizophrenia and is modestly effective for smoking cessation in patients with schizophrenia. The relapse rate is high after treatment discontinuation. PMID- 15876900 TI - Inhibition of CYP2D6 activity by bupropion. AB - The purpose of this study was to assess the effect of bupropion on cytochrome P450 2D6 (CYP2D6) activity. Twenty-one subjects completed this repeated-measures study in which dextromethorphan (30-mg oral dose) was administered to smokers at baseline and after 17 days of treatment with either bupropion sustained-release (150 mg twice daily) or matching placebo. Subjects quit smoking 3 days before the second dextromethorphan administration. To assess CYP2D6 activity, urinary dextromethorphan/dextrorphan metabolic ratios were calculated after an 8-hour urine collection. Thirteen subjects received bupropion, and 8 received placebo. In those receiving active medication, the dextromethorphan/dextrorphan ratio increased significantly at the second assessment relative to the first (0.012 +/- 0.012 vs. 0.418 +/- 0.302; P < 0.0004). No such change was observed in those randomized to placebo (0.009 +/- 0.010 vs. 0.017 +/- 0.015; P = NS). At baseline, all subjects were phenotypically extensive CYP2D6 metabolizers (metabolic ratio <0.3); after treatment, 6 of 13 subjects receiving bupropion, but none of those receiving placebo, had metabolic ratios consistent with poor CYP2D6 metabolizers. Bupropion is therefore a potent inhibitor of CYP2D6 activity, and care should be exercised when initiating or discontinuing bupropion use in patients taking drugs metabolized by CYP2D6. PMID- 15876901 TI - Efficacy of buspirone in the treatment of opioid withdrawal. AB - In an attempt to develop a new opiate detoxification approach, the authors assessed the efficacy of buspirone in the treatment of acute heroin withdrawal. Buspirone, a drug interacting with the serotonergic system, was selected because there is evidence that a decrease in serotonergic neurotransmission may be involved in opiate withdrawal symptoms. Twenty-nine hospitalized heroin addicts were randomized to 4 groups: (1) placebo; (2) methadone; (3) buspirone 30 mg daily; (4) buspirone 45 mg daily. The double-blind trial started in all patients with a 5-day methadone stabilization period ending with a 30-mg dose. This was followed from days 6 through 12 by placebo in group 1 and by a methadone taper in group 2. Because of its delayed action, buspirone was started on day 1 in groups 3 and 4 and was continued, after methadone discontinuation, through day 12. On day 13, drugs and placebo were discontinued and patients were observed through day 14. Withdrawal symptoms were assessed with the "Subjective Opiate Withdrawal Scale" (SOWS) and the "Objective Opiate Withdrawal Scale" (OOWS). The SOWS and OOWS scores were significantly higher in the placebo group than in the methadone, buspirone 30 mg, and buspirone 45 mg groups. There were no significant differences in SOWS or OOWS scores when the methadone group was compared with each of the two buspirone groups or when the two buspirone groups were compared with one another. In conclusion, buspirone, a nonopiate drug with no abuse potential, a safe side effect profile and no withdrawal symptoms, at doses of 30 and 45 mg, was as effective as a methadone taper in alleviating the withdrawal symptoms of heroin addicts stabilized for 5 days with, and then withdrawn from, methadone. The use of buspirone could be particularly helpful in outpatient settings where the duration of the methadone taper recommended for detoxification can be lengthy. PMID- 15876902 TI - The effect of citalopram adjunctive treatment added to atypical antipsychotic medications for cognitive performance in patients with schizophrenia. AB - Cognitive enhancement in patients with schizophrenia is a major treatment priority. Because serotonergic approaches have been suggested as a possible mechanism to enhance cognition and many patients with schizophrenia are treated with selective serotonin reuptake inhibitor antidepressants, we evaluated a serotonin reuptake inhibitor, citalopram, as adjunctive therapy to atypical antipsychotic treatment for its cognitive enhancing effects in schizophrenic patients. Nineteen schizophrenic patients were treated in a randomized, placebo controlled, crossover-designed 24-week study. In phase 1, subjects were randomized equally to 40 mg of citalopram or placebo and were evaluated prior to initiation of pharmacotherapy and at the end of phase 1 (after 12 weeks of treatment with double-blind agent). At the beginning of phase 2, subjects were crossed over to the other treatment and subsequently assessed after 12 weeks of treatment for symptom severity and cognitive performance. There were no statistically significant differences between citalopram 40 mg/d and placebo treatment on any clinical or cognitive measures. These results indicate that citalopram adjunctive treatment to atypical antipsychotics produces no significant cognitive improvement in patients with schizophrenia. Because the subjects in this study were all treated with atypical antipsychotics, it is possible that the pharmacologic profiles of atypical antipsychotic medications at serotonin receptors may have complicated the effects of citalopram augmentation. Further research on alternative serotonergic approaches to cognitive enhancement in schizophrenia is warranted. PMID- 15876903 TI - Hyperforin plasma level as a marker of treatment adherence in the National Institutes of Health Hypericum Depression Trial. AB - BACKGROUND: A previously reported clinical trial of Hypericum perforatum (St John's wort) in depression did not demonstrate efficacy. We assessed treatment adherence by measuring plasma hyperforin and evaluated the possible impact of adherence on study results. METHODS: Outpatients with major depression (N = 340) were randomized to an 8-week trial of H. perforatum (900-1500 mg/d), sertraline (50-100 mg/d) as active comparator, or placebo. Plasma was available from 292 patients (86% of randomized). Samples from the placebo and H. perforatum groups were assayed for hyperforin, and samples from the sertraline group for sertraline/N-desmethyl-sertraline. RESULTS: Of the 104 patients randomized to placebo, 18 (17%) had detectable plasma hyperforin. Of the 97 patients randomized to H. perforatum, 17 (17%) had no detectable plasma hyperforin. All the assayed sertraline patients (N = 91) had plasma sertraline/N-desmethyl-sertraline. The clinical trial conclusions remained unchanged when only patients with plasma assay consistent with random assignment were included in the analyses. CONCLUSIONS: One of every 6 patients assigned to placebo had plasma hyperforin, and 1 of every 6 patients assigned to H. perforatum had no detectable plasma hyperforin. The finding underscores the difficulty of enforcing treatment adherence in clinical trials of preparations that are readily available in the community. PMID- 15876904 TI - The association of weight gain and olanzapine plasma concentrations. AB - Atypical antipsychotics, including olanzapine, have been associated with clinically significant weight gain in some patients. The purpose of this study was to determine if weight gain was associated with increasing plasma concentrations during olanzapine treatment in subjects with schizophrenia. This study included 39 acutely ill subjects with schizophrenia, schizoaffective disorder, or schizophreniform disorder (DSM-III-R or DSM-IV). Assessments included the Brief Psychiatric Rating Scale (BPRS), the Scale for Assessment of Negative Symptoms (SANS), and weight measurements. Olanzapine was titrated to a dose of 5 to 20 mg/d for 2 to 6 weeks. A 24-hour plasma concentration was obtained after 6 weeks of treatment. Analysis using a receiver operator characteristic curve identified a threshold dose-weighted plasma concentration of 20.6 ng/mL being associated with an increased likelihood of clinically significant weight gain (> or =7% baseline weight) during olanzapine treatment. The associations remained significant after adjusting for age, gender, baseline body mass index, baseline symptom severity, and symptom improvement (OR = 10.1; 95% CI, 1.3-75.0; P = 0.024). Similar analysis determined that a threshold olanzapine dose of 13.3 mg/d was associated with > or =7% weight gain. However, after adjusting for potential confounders, the results did not remain significant. The association of weight gain with plasma concentrations during treatment with olanzapine may support the utilization of plasma drug concentration as a marker for antipsychotic-induced weight gain in the treatment of schizophrenia. PMID- 15876905 TI - Association between early and rapid weight gain and change in weight over one year of olanzapine therapy in patients with schizophrenia and related disorders. AB - Weight gain is an important issue in the use of atypical antipsychotics, including olanzapine. A retrospective analysis of patterns of weight gain and possible covariates was performed for 1191 patients diagnosed with schizophrenia or schizoaffective disorder who were treated with olanzapine for up to 52 weeks. Patients were dichotomized into 2 main groups according to the percentage of body weight gained during the first 6 weeks of treatment with olanzapine: (1) patients who gained > or =7% of their body weight (Rapid Weight Gain Group [RWG]), and (2) patients who lost weight, gained no weight, or gained <7% of their body weight (Nonrapid Weight Gain Group [NRWG]). Results demonstrated that approximately 15% of the patient population showed rapid increases in weight (RWG group), whereas 85% of patients gained weight more slowly or not at all (NRWG group). Patients in the RWG group gained an average of 4% of their body weight (approximately 4-7 lb) within the first 2 weeks of treatment with olanzapine. Furthermore, patients in the RWG group were younger, had a lower baseline body mass index, were more likely to report an increase in appetite, and showed a more robust clinical response compared with patients in the NRWG group. Over the course of 52 weeks, patients in the RWG group gained significantly more weight and reached a higher plateau for mean weight increase at 38 weeks compared with the mean increase observed for patients in the NRWG group. By measuring the weight of patients during the first few weeks of olanzapine treatment and by assessing changes in appetite, clinicians may be able to identify those patients at risk for substantial weight gain. PMID- 15876906 TI - Bone mineral density and its relationship to prolactin levels in patients taking antipsychotic treatment. AB - Antipsychotic treatment is frequently associated with elevated prolactin levels. Raised prolactin levels have been linked with osteoporosis. The objective of this study is to determine whether patients taking antipsychotics show reduced bone mineral density (BMD), and whether this is associated with prolactin levels. BMD (standardized as z scores) was compared using dual x-ray absorptiometry of the lumbar spine and hip in patients taking antipsychotics (n = 102, mean age: 46.0, SD: 13.1, 47% male, median treatment duration: 3.0 years) to matched reference controls. Levels of prolactin, markers of bone metabolism, and risk factors for osteoporosis were measured. Mean BMD was not significantly reduced, other than the total spine score for black males (mean z score: -0.88, P = 0.00001). BMD was correlated with body mass index but there was no correlation with prolactin. BMD was not correlated with prolactin levels and showed no clinically significant reduction. The low BMD in black males warrants further investigation. PMID- 15876907 TI - Activity of citalopram on adenosine and serotonin circulating levels in depressed patients. AB - Citalopram is a selective serotonin reuptake inhibitor used in the treatment of depression. Recent investigations have shown that it reduces in rat brain the release of excitatory amino neurotransmitters acid glutamate and aspartate by the involvement of the inhibitory neuromodulator adenosine. In this study, we described citalopram and serotonin levels in plasma and platelets, as well as plasma adenosine levels, in depressive patients during acute and chronic administration of citalopram. Twelve patients affected by Major Depression (DSM IV) received a single oral dose of citalopram in the morning, 5 mg in the first 5 days, 10 mg from the 6th to the 10th day, and 20 mg from the 11th to the 40th day. Blood samples for citalopram, serotonin, and adenosine were collected at Time 0 and 4, 12 and 24 hours after drug administration on the first day of citalopram 5 mg, and on the first and the last day of citalopram 20 mg. Citalopram, serotonin, and adenosine concentrations in plasma increased after citalopram administration, and the highest levels were observed on the last day of treatment. Citalopram was detectable in platelets with concentrations showing a time variation similar to plasma values. Serotonin levels in platelets decreased after drug administration, reaching the lowest values on the last day of treatment. PMID- 15876908 TI - An open trial of naltrexone in the treatment of depersonalization disorder. AB - Depersonalization disorder (DPD) remains one of the few disorders in modern psychiatry for which no treatments are established that are even partially effective, whether pharmacological or psychotherapeutic. Depersonalization disorder is a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition dissociative disorder characterized by a pervasive subjective sense of unreality and detachment with intact reality testing. Two recent controlled medication trials, one with lamotrigine and one with fluoxetine, failed to show efficacy. There is some evidence for dysregulation of endogenous opioid systems in depersonalization, and a few studies have suggested that opioid antagonists may have efficacy in the treatment of dissociation and depersonalization symptoms. In this prospective open treatment trial, 14 subjects were recruited and treated with naltrexone for 6 weeks to a maximum dose of 100 mg/d (first 7 subjects) or 10 weeks to a maximum dose of 250 mg/d (next 7 subjects). Mean naltrexone dose was 120 mg/d. There was an average 30% reduction of symptoms with treatment, as measured by 3 validated dissociation scales. Three patients were very much improved, and 1 patient was much improved with naltrexone treatment. These findings are potentially promising in a highly treatment-refractory disorder for which no treatment guidelines exist and warrant a randomized controlled trial. PMID- 15876909 TI - Effect of study criteria on recruitment and generalizability of the results. AB - OBJECTIVE: Clinical trials are indispensable to drug approval process. This research examined the effect of a specific study criteria on recruitment and generalizability of the results. METHODS: The following were reviewed: (a) the usual inclusion and exclusion criteria for the antipsychotic trials performed at the Institute; (b) epidemiologic data, to determine the effect of study criteria on the target population; and (c) the recruitment procedures/strategies used to identify potential candidates. A survey was conducted to determine the percentage of schizophrenic patients in a conventional outpatient psychiatric clinic conforming to the usual enrollment criteria for antipsychotic trials. RESULTS: Intensive recruitment efforts in a general population of 3.6 million would have been expected to yield only 632 eligible subjects out of 36,000 suffering from schizophrenia. Out of 632, only 50 contacted the research site after an intensive recruitment effort. From those 50, 30 were excluded during a telephone interview. Of the 20 remaining, 6 were excluded for a variety of reasons during a face-to face interview. Thus, only 14 subjects out of a population of 3.6 million met the study criteria. CONCLUSIONS: These results emphasize the rarified nature of patients-volunteers who enter a clinical trial. Inclusion and exclusion study criteria can severely restrict the number of eligible subjects, dictate recruitment strategies, and in turn affect generalizability of the results. PMID- 15876910 TI - Recurrence of clozapine-induced agranulocytosis. PMID- 15876911 TI - Double-blind, placebo-controlled, crossover trial of clozapine plus glycine in refractory schizophrenia negative results. PMID- 15876912 TI - The influence of olanzapine versus risperidone on facial expression of emotions in schizophrenia--preliminary results of a facial electromyogram study. PMID- 15876913 TI - Effective adjunctive use of pergolide with quetiapine for cognitive impairment and negative symptoms in schizophrenia. PMID- 15876914 TI - The effects of bupropion on heart rate variability in healthy volunteers. PMID- 15876915 TI - Antidepressant pharmacoepidemiology in a general population sample. PMID- 15876916 TI - Levofloxacin-related seizure activity in a patient with Alzheimer's disease: assessment of potential risk factors. PMID- 15876917 TI - Perospirone improves food refusal in elderly patients with mental disorders. PMID- 15876918 TI - Epidemiology of pertussis. AB - The World Health Organization recommended that a pertussis incidence of <1 case per 100,000 population be achieved in Europe by 2000. Available data indicate that this goal has generally not been achieved, and the incidence is actually rising in some countries. Understanding the reasons for this increased incidence may lead to better global control of pertussis. In the majority of countries where pertussis is a notifiable disease, a case-based national surveillance system is in place. However, different case definitions, methods of diagnosis and reporting and surveillance systems make direct intercountry comparisons difficult, and pertussis is not a statutory notifiable disease in every country. Nevertheless the general consensus is that reported incidences are probably considerably lower than the actual incidence of pertussis; underreporting is common. Prolonged cough may be the only clinical feature in adolescents or adults, who may present for diagnosis late (precluding laboratory confirmation) or not at all. When they do present, their condition is often misdiagnosed because, in part, clinicians continue to perceive pertussis as a childhood disease. Despite underreporting, an increased incidence of infant, adolescent and adult pertussis has been observed worldwide since the introduction of widespread vaccination. This is of concern because adolescents and adults have been identified as a source of transmission of pertussis to very young infants who are unimmunized or partially immunized and thus more vulnerable to disease-related complications and higher mortality. In recent years, acellular pertussis vaccines have been incorporated into the immunization schedules of many developed countries, gradually replacing whole cell vaccines. Dosing schedules vary between countries, although primary immunization with 3 doses of the pertussis vaccine within the first 6 months of life exists in most countries. Only Australia, Austria, Canada, France and Germany have incorporated an adolescent booster dose into their current immunization schedules, in recognition of the rising incidence of pertussis in adolescents and adults. Despite high coverage rates for primary immunization in infants and children, pertussis continues to be a global concern, with increased incidence widely noted. This global epidemiologic summary highlights differences worldwide in pertussis reporting, incidence and approaches to prevention. It underscores a general shift in the age distribution of pertussis toward older groups. Understanding the link between these observations may lead to better informed global control strategies, especially those pertaining to immunization schedules and use of pertussis vaccine. PMID- 15876919 TI - Pertussis sources of infection and routes of transmission in the vaccination era. AB - Vaccination against pertussis has resulted in reduction of the infection pressure of Bordetella pertussis (partial herd immunity), but the circulation of B. pertussis has persisted as a consequence of waning of vaccine-induced and naturally acquired immunity. An increase in the reported incidence of B. pertussis infection in older children, adolescents and young adults has been noted, resulting in a perceived resurgence of the disease in these age groups. Regardless of whether this resurgence is real or not, older groups are increasingly recognized as playing an important role in transmitting B. pertussis infection to incompletely immunized infants, in whom pertussis disease continues to cause severe and fatal illness, albeit at much lower levels than in the prevaccine era. Several studies have suggested that mothers, in particular, are a significant source of infection for infants. Adolescents, grandparents and health care workers can also play a role. By contrast, most adolescents acquire the infection from schoolmates and friends, whereas for adults the main sources are children and work colleagues. Furthermore teachers, child care workers and health care workers could be at increased risk of being exposed to, and transmitting, B. pertussis infection. Current immunization strategies inadequately control the circulation of B. pertussis, in part because of suboptimal adherence to current pediatric immunization guidelines. In addition to efforts to improve pertussis immunization rates in children, the expansion of pertussis immunization to target specific groups should be considered. Besides reducing morbidity in the targeted groups, these strategies could decrease the residual burden of pertussis morbidity and mortality in infants. PMID- 15876920 TI - Defining pertussis epidemiology: clinical, microbiologic and serologic perspectives. AB - In the past decade, numerous sources have noted an increase in reported pertussis in highly immunized populations. This has been accompanied by a perceived change in disease epidemiology, characterized by a significant increase in reported pertussis incidence among adolescents and adults. In populations where children are routinely immunized, adolescents and adults now constitute the main source of infection in infants. However, a range of factors makes delineation of these epidemiologic trends difficult. Reported cases of pertussis represent only a fraction of the actual number of Bordetella pertussis symptomatic infections, because underconsulting, underrecognition and underdiagnosis are widespread and are a particular problem in adolescents and adults. Possible explanations for failure to diagnose pertussis include the heterogeneity in pertussis disease expression and low physician awareness and index of suspicion. Consequently defining pertussis from a clinical perspective is difficult, and this is reflected by a lack of consistency between case definitions. Although case definitions for specific circumstances have been established by the World Health Organization and the United States Centers for Disease Control, these are not universally useful, making intercountry comparisons and global evaluation difficult. Less-than-optimal and poorly performed laboratory tests, or their lack of availability, also make confirmation of B. pertussis infection difficult. To overcome these problems, clinical case definitions should be standardized for outbreak and endemic situations. Rapid, easy-to-use and inexpensive laboratory diagnostic techniques also must be made available and be widely implemented. In particular, polymerase chain reaction and single serum serology are 2 techniques that should be more widely adopted. PMID- 15876921 TI - Summary: epidemiology of pertussis. PMID- 15876922 TI - Health burden of pertussis in infants and children. AB - In countries with high rates of vaccination against pertussis, the incidence of this disease has decreased dramatically compared with the prevaccine era. However, pertussis still occurs in these countries, and severe morbidity and mortality are greatest among infants, particularly those who are unimmunized or incompletely immunized. Pertussis in older children and adults is perceived by many as being a mild disease, but it is a significant health burden in persons of all ages. Infants with pertussis experience the highest rates of hospitalization, complications and death. Severe complications include pneumonia, encephalopathy and meningoencephalitis. In addition, infants may experience weight loss, bronchitis, otitis media, apnea, cyanosis, inguinal hernia and rectal prolapse. It is essential to explore methods to prevent disease transmission to infants in the months before they complete their primary immunization series. The Global Pertussis Initiative was established to assess the true health burden of pertussis in infants and to suggest strategies to combat transmission and infection with Bordetella pertussis, which remains a significant public health concern. PMID- 15876923 TI - Health burden of pertussis in adolescents and adults. AB - Pertussis in adolescents and adults is common, endemic, and epidemic worldwide, and its incidence is reportedly increasing. Although a number of individuals suffer only a mild cough, many others have symptoms typical of pertussis, causing prolonged cough illness, frequent use of health care resources, missed work and a variety of complications. Symptoms experienced by adolescents and adults include sleep disturbance, weight loss, pharyngeal discomfort, influenza-like symptoms, sneezing attacks, hoarseness, sinus pain, headaches and sweating attacks. Even when symptoms are typical of pertussis, the diagnosis is often not considered in adolescents and adults because of a low awareness of the disease in these age groups. Contrary to common perceptions, complications of pertussis, including some that are serious, are not infrequent in adolescents and adults. These include urinary incontinence, rib fracture, pneumothorax, inguinal hernia, aspiration, pneumonia, seizures and otitis media. Despite underreporting, hospitalization of adults and adolescents does occur. Many believe that adolescents and adults are the groups most commonly infected with pertussis and are now the major source of contagion to infants and young children. Because of the considerable health burden, there is a need for improved vaccination strategies to prevent disease in adolescents and adults and to reduce the risk of transmission to vulnerable infants. PMID- 15876924 TI - Economic burden of pertussis and the impact of immunization. AB - Although routine use of vaccines has diminished the incidence of pertussis disease, it has not eliminated the pathogen. Epidemiologic data confirm that pertussis remains a significant health problem in all age groups. Disease burden is highest in infants, in whom pertussis disease frequently leads to severe complications and mortality, although it is also a significant health burden in adolescents and adults, in whom the reported incidence of pertussis is increasing. The Global Pertussis Initiative reviewed the literature to find data that express the economic impact of this health burden and to review economic evaluations of pertussis immunization. Although only limited data on the direct and indirect costs of pertussis are available, they suggest that it poses a significant economic burden and indicate that the direct medical costs of pertussis depend on the rate of hospitalization and the severity of complications, and are highest in infants. The indirect costs of pertussis also appear to be considerable, particularly among adults, in whom the disease reduces work productivity, because of either personal illness or child care responsibilities. Several health economic models on the cost effectiveness of childhood immunization strategies have been published, and although constrained by missing data, have generally found childhood immunization strategies to be cost-effective. Economic analyses of adolescent and adult immunization strategies have also been conducted, but the findings of these studies have been inconsistent. The most recent evaluations, using much higher estimates of incidence than reported previously, suggest that immunization of adolescents and specific adult subgroups may be cost-effective. The literature review confirmed that the economic burden of pertussis is substantial, but there are gaps in existing information. In the short term, further economic analyses are required, particularly of adolescent and adult immunization. More importantly, collection of primary epidemiologic and economic data should be undertaken in parallel. Despite the existing gaps in data, further research using the most current data should facilitate decisions on new vaccination strategies by describing conditions for favorable results and quantifying the margin of uncertainty. PMID- 15876925 TI - Aims, scope and findings of the global pertussis initiative. PMID- 15876926 TI - Summary: health and economic burden of pertussis. PMID- 15876927 TI - Duration of immunity against pertussis after natural infection or vaccination. AB - Despite decades of high vaccination coverage, pertussis has remained endemic and reemerged as a public health problem in many countries in the past 2 decades. Waning of vaccine-induced immunity has been cited as one of the reasons for the observed epidemiologic trend. A review of the published data on duration of immunity reveals estimates that infection-acquired immunity against pertussis disease wanes after 4-20 years and protective immunity after vaccination wanes after 4-12 years. Further research into the rate of waning of vaccine-acquired immunity will help determine the optimal timing and frequency of booster immunizations and their role in pertussis control. PMID- 15876928 TI - Role of maternal pertussis antibodies in infants. AB - Pertussis remains a serious infection in young infants. Most deaths occur in the first 3 months of life, before administration of the first dose of pertussis vaccine. Pertussis antibodies are transferred from mother to infant; but because of the lack of serologic correlates of protection, it is difficult to determine the proportion of infants born with a protective concentration of maternal antibodies. Indirect evidence suggests that maternal antibodies provide short lived protection against fatal pertussis. It is hoped that the protection of young infants could be enhanced by maternal or neonatal vaccination. The possibility of protecting young infants against pertussis by immunizing their mothers during pregnancy was investigated in the 1930s and 1940s; no further studies have been published since. Recent animal and human studies have provided evidence that neonatal immunization with acellular pertussis vaccine can efficiently prime T and B cells and act as a basis for future immune response. The limited data on neonatal and maternal pertussis immunization are promising and call for further research to reduce the vulnerability of young infants to pertussis disease. PMID- 15876929 TI - Use of antibiotics in the prevention and treatment of pertussis. AB - Antibiotics are commonly used for treating confirmed cases of pertussis and also for disease prevention in outbreak situations, and there is little evidence of antibiotic resistance of Bordetella pertussis. The most commonly used antibiotic is erythromycin, but the associated side effects limit compliance and therefore efficacy. Other antibiotics, such as clarithromycin and azithromycin, have been shown to be at least as effective as erythromycin in preventing and treating pertussis, and they also have fewer side effects, which improves compliance. This article outlines the use of different antibiotics in pertussis management and their effect on preventing disease transmission and reducing disease severity and duration. PMID- 15876930 TI - Potential strategies to reduce the burden of pertussis. AB - Pertussis continues to be a significant cause of morbidity and mortality among nonimmunized young infants. Although the inception of childhood pertussis immunization programs has significantly reduced the occurrence of the disease in children, waning vaccine-induced immunity permits the disease to affect adolescents and adults, who in turn transmit the disease to unimmunized or incompletely immunized infants. The Global Pertussis Initiative brought together experts from 17 countries around the world to evaluate strategies to improve disease control. Seven strategies were considered: (1) universal adult immunization; (2) selective immunization of mothers and close family contacts of newborns; (3) selective immunization of health care workers; (4) selective immunization of child care workers; (5) universal immunization of adolescents; (6) preschool booster at 4-6 years of age; and (7) reinforcement and/or improvement of current infant and toddler immunization strategies. Because immunization programs vary widely from country to country, no single strategy is likely to be appropriate for all. Moreover it would be helpful to have additional data to support the strategies and provide a better understanding of the disease so that new approaches can be monitored effectively. However, certain steps can be taken now to reduce the incidence of pertussis. PMID- 15876931 TI - The global pertussis initiative: process overview. AB - In 2001, the Global Pertussis Initiative was established as a scientific forum to analyze the status of pertussis globally and to evaluate various immunization strategies to improve disease control. Thirty-seven multidisciplinary experts from 17 countries participated. The initiative was conducted in 3 stages: assessment of the international epidemiology, diagnosis, health and economic burden, and prevention and treatment of pertussis; evaluation and prioritization of 7 immunization strategies to address the problems; and identification of solutions to surmount potential barriers to the implementation of these strategies. A health-economic model, created for the Initiative, analyzed the cost-effectiveness of the selected immunization strategies. Discussion, debate, and consensus were facilitated via a roundtable meeting, teleconferences and use of a closed, interactive website, allowing the participants to share data, knowledge, experience, and opinion. This article describes the processes undertaken to conduct the Initiative. PMID- 15876932 TI - Pertussis immunization of adolescents in the United States: an economic evaluation. AB - The incidence of reported pertussis has increased during the past decade and poses a growing health and economic burden in developed countries, despite high rates of primary vaccination. Administration of a booster dose of acellular pertussis vaccine to adolescents may help reduce this burden, not only by reducing infections in vaccinated individuals but also by reducing transmission of Bordetella pertussis to other individuals, particularly infants. An epidemiologic model was created to assess the health and economic impact of implementing a program of routine acellular pertussis immunization in adolescents 11-18 years of age in the United States, considering both the reduction in cases in those vaccinated and among the unvaccinated population (due to herd immunity). Inputs for the base case were defined according to information derived from published literature and were supplemented by estimates provided by members of the Global Pertussis Initiative. Both direct and indirect costs were included (in 2002 US dollars) using U.S. data. Outcomes were evaluated over the lifetime of a cohort of potential adolescent vaccine candidates. Because of uncertainty in many of the inputs, extensive sensitivity analyses were conducted. With 80% vaccination coverage of adolescents and a 20% reduction of other cases because of herd immunity, >68,000 cases and 41 pertussis-related deaths would be avoided in the subsequent 10 years by routine administration of acellular pertussis boosters to a single cohort of adolescents in the United States. This strategy would be cost-effective, incurring from 6000 US dollars to 22,000 US dollars per life-year gained. The level of herd immunity attained and the true incidence of pertussis are critical determinants of cost effectiveness, as is the duration of immunity resulting from immunization. The cost of immunization and the discount rate also play a role. Although there is considerable uncertainty surrounding key inputs, the results indicate that the conditions required for adolescent immunization to be economically warranted are realistic. PMID- 15876933 TI - Pertussis immunization in the global pertussis initiative North American region: recommended strategies and implementation considerations. AB - In North America, children currently receive 5 doses of a combined diphtheria tetanus-acellular pertussis vaccine between the ages of 2 months and 6 years. Although this schedule has reduced the incidence of childhood pertussis, it has not led to the development of herd immunity in the total population, largely because pertussis immunity wanes with time. The time course over which immunity wanes is uncertain; however, high pertussis antibody titers in adolescents and adults indicate unrecognized infection in these groups. There is evidence that this group serves as a source of infection for young infants who are not fully immunized. Therefore, of the potential strategies reviewed by the North American Global Pertussis Initiative group, universal adolescent immunization would in theory reduce the risk of pertussis in this age group and may reduce transmission to young infants. However, because immunity probably wanes at the same rate in adolescents and children, the burden of disease will likely shift to older age groups, including young adults (parents of vulnerable infants). Therefore the ideal would be immunization of adolescents and adults, particularly those who are in contact with young infants. Adolescent immunization is already recommended in Austria, France, Germany and Canada, and participants in the Global Pertussis Initiative recommend that this strategy be implemented across North America with a view to eventually extending immunization to include adults. The final decision to implement such a strategy will depend on pertussis surveillance studies and analysis of the effectiveness and tolerability of adolescent and adult pertussis immunization as well as program considerations related to feasibility and economics. PMID- 15876934 TI - Pertussis immunization in the global pertussis initiative European region: recommended strategies and implementation considerations. AB - Approaches to pertussis diagnosis, surveillance and immunization vary widely across Europe. Nonetheless most countries report high levels of vaccine coverage in infants and toddlers, and significant reductions in infant morbidity and mortality have been achieved. As a consequence of the effective protection of infants and toddlers, the absolute incidence of pertussis has substantially decreased, but the relative proportion of older age groups, adolescents and adults in particular, has increased. These groups, however, are a relevant source of infection of unimmunized or incompletely immunized infants. In addition to efficient childhood vaccination, other approaches to pertussis immunization are required. Among the various strategies evaluated, 3 were recommended by the European participants in the Global Pertussis Initiative that might be adapted to each country's specific needs: the reinforcement of implementation of current schedules, the addition of an extra dose of vaccine to current immunization schedules and the selective immunization of health care workers, which is already included in a European Commission directive. The main barriers to the acceptance of these strategies are low awareness of pertussis in immunized populations, poor recognition of the disease in adults and adolescents, lack of standardized diagnostic criteria and poor access to laboratory confirmation of the diagnosis. These obstacles have led to underreporting of pertussis and an underestimation of the disease burden. Actions to overcome these issues are crucial to the implementation of new or improved immunization strategies to combat pertussis in Europe. PMID- 15876935 TI - Pertussis immunization in the global pertussis initiative international region: recommended strategies and implementation considerations. AB - Despite widespread immunization programs in most countries, pertussis disease continues to be a threat to public health. In particular, there has been a resurgence of pertussis disease in older children, adolescents and adults, creating a reservoir of infection, which poses a significant threat to infants who are either unimmunized or incompletely immunized. Global Pertussis Initiative participants from Argentina, Australia, Brazil and Japan considered the relative merits of several strategies to reduce the burden of pertussis disease and suggested strategies that might be implemented in these countries. Infants in these countries receive an initial course of 3 doses of vaccine in the first year of life followed by a fourth dose in the second year. Only children in Japan are not given a preschool booster (age 3-5 years). Of the strategies considered, the addition of a preschool booster is therefore a priority in Japan to overcome the problem of waning vaccine-induced immunity to pertussis in school children. Waning immunity also affects adolescents; Australia introduced an adolescent booster in 2003, and the addition of a booster in this age group was suggested for Argentina and Japan. Immunization of new mothers and other close contacts of young infants, such as child care and health care workers, might be appropriate in Australia in the future. Argentina also suggested a future possibility of immunizing health care and child care workers. Obstacles to new immunization strategies include poor access to standardized laboratory diagnostic techniques, inadequate resources to fund new immunization programs, low awareness of pertussis disease in adults and adolescents and inadequate surveillance techniques to assess the full extent of the problems caused by pertussis or the impact new vaccination strategies might have. PMID- 15876936 TI - Controlling pertussis: considerations for the future. PMID- 15876937 TI - Decreasing hospital-associated rotavirus infection: a multidisciplinary hand hygiene campaign in a children's hospital. AB - OBJECTIVE: Hand hygiene is an effective means of preventing hospital-associated infection, but compliance among health care workers is poor. Few studies aimed at increasing hand hygiene in the hospital setting have shown sustained improvement and concurrent decreases in hospital-associated infections, and even fewer have been performed in the pediatric setting. METHODS: We implemented a hand hygiene program with the hopes of improving hand hygiene and decreasing hospital associated rotavirus infection rates. A multidisciplinary group developed a hospital supported, house-wide campaign. Opportunities for hand hygiene were observed during 5 periods. The frequency of hospital-associated rotavirus infection was tracked over time by review of laboratory records. Correlates of hand hygiene were investigated with the use of multivariate logistic regression. RESULTS: Overall hand hygiene compliance improved from 62% in period 1 to 81% in period 5 (P < 0.001). Soap and water was the most common method for practicing hand hygiene, and alcohol hand gel use increased from 4% to 29% between the first and last observation periods (P < 0.001). The rate of hospital-associated rotavirus infection decreased from 5.9 episodes per 1000 discharged patients in 2001 to 2.2 episodes per 1000 discharged patients in 2004 (P = 0.01). Period of observation, hospital ward, type of care provider and type of care performed were all independently associated with hand hygiene (adjusted P < or = 0.02 for all). CONCLUSION: Improving hand hygiene is an important goal for health care institutions. These data can be useful for development of interventions aimed at improving hand hygiene. PMID- 15876938 TI - Antibiotic-resistant Helicobacter pylori strains in Portuguese children. AB - BACKGROUND: Data concerning the effectiveness of Helicobacter pylori eradication regimens based in antibiotic susceptibility testing are scanty in children. AIMS: To identify the prevalence of antibiotic resistance in H. pylori strains isolated from Portuguese children in 1999-2003; to evaluate eradication rate after antibiotic susceptibility testing-based treatment; and to identify factors associated with resistance and eradication outcome. METHODS: Included were 109 children with a gastric biopsy culture positive for H. pylori. First treatment (amoxicillin, omeprazole and clarithromycin or metronidazole) was guided by susceptibility testing (E test), and eradication was assessed by [C]urea breath test. RESULTS: Strains were susceptible to amoxicillin and tetracycline; 39.4% were resistant to clarithromycin, 16.5% to metronidazole and 4.5% to ciprofloxacin. No significant association was found between resistance and sex, age, clinical status, gastritis scores, H. pylori density scores and genotype. Clarithromycin resistance was significantly associated with European origin [odds ratio (OR), 3.9], previous H. pylori empiric therapy (OR 2.8) and amoxicillin minimal inhibitory concentration, > or =0.016 (OR 6.0). Eradication rate after susceptibility-based treatment was 74.7% (59 of 79; 95% confidence interval, 65.9 82.9), and a significant association was found between eradication failure and presence of resistance to 1 or more antibiotics (P < 0.05). CONCLUSIONS: The prevalence of H. pylori antibiotic resistance was high in the studied population. The modest therapeutic success of clarithromycin and metronidazole susceptibility based regimens suggests that in addition to resistance, other factors may be involved. The need of susceptibility-based treatment studies in children and of antimicrobial resistance surveillance in high prevalence areas for H. pylori are emphasized. PMID- 15876939 TI - Quantitative and qualitative antibody response to pneumococcal conjugate vaccine among African human immunodeficiency virus-infected and uninfected children. AB - OBJECTIVE: To determine the quantitative and qualitative antibody responses to a pneumococcal conjugate vaccine (PnCV) in human immunodeficiency virus (HIV) exposed infected and uninfected children. METHODS: Children were randomized to receive either a PnCV or placebo at 6, 10 and 14 weeks of age. PnCV serotype specific antibody concentrations were measured by a standard enzyme immunoassay (EIA) and a 22F modified EIA (22F EIA) on single serum samples drawn at 21-42 days post-dose 3. Functional activities of the serotype-specific antibody to serotypes 6B, 19F and 23F were measured with an opsonophagocytic assay (OPA). RESULTS: The geometric mean antibody concentrations (GMC) were similar in HIV infected and HIV-uninfected PnCV recipients for 7 of the 9 vaccine serotypes. In placebo recipients, the GMCs were significantly higher in HIV-infected than in uninfected children for 7 of the serotypes. In HIV-infected PnCV recipients, the GMCs were lower for 5 of the serotypes in children with severe acquired immunodeficiency syndrome than in children who were asymptomatic or mildly symptomatic with acquired immunodeficiency syndrome. HIV-infected PnCV recipients were less likely to have measurable functional antibody (OPA titer > or =1/8) to all 3 studied serotypes (6B, 19F and 23F) than in HIV-uninfected children. HIV infected children required a higher concentration of anticapsular antibody to achieve 50% of the maximum uptake of labeled Streptococcus pneumoniae in the OPA assay than HIV-uninfected children for 2 of the 3 serotypes, although this was significant only for serotype 6B (P = 0.0005). CONCLUSION: HIV-infected children have similar quantitative antibody responses but poorer qualitative antibody responses to the PnCV. PMID- 15876940 TI - GB virus C infection in children with perinatal human immunodeficiency virus infection. AB - BACKGROUND: GB virus C (GBV-C) infection occurs in 20-40% of human immunodeficiency virus (HIV)-infected adults, and coinfection is associated with improved HIV disease outcome. METHODS: To determine the prevalence of GBV-C infection in children who were perinatally infected with HIV, we conducted a cross-sectional prevalence survey in a cohort of perinatally infected HIV positive children selected from a large, multicenter observational protocol. A blood specimen was obtained and tested for GBV-C viremia with the use of a qualitative GBV-C RNA assay and screened for past GBV-C infection with enzyme linked immunosorbent assay to detect antibodies to the GBV-C envelope protein E2 (E2 Ab). RESULTS: The 354 children who participated in the substudy were relatively healthy, with a median CD4 of 784 cells/mm and median HIV-1 viral load of 1055 copies/mL. The prevalence of GBV-C viremia was 20 of 353 or 5.7% (95% confidence interval, 3.5-8.6%), and the prevalence of E2 Ab was 12 of 354 or 3.4% (95% confidence interval, 1.8-5.8%). GBV-C viremic patients were older than patients without past GBV-C infection (median age, 12.8 years versus 10.7 years). Median CD4 lymphocyte counts were highest in subjects without GBV-C infection and lowest in those with E2 Ab. CONCLUSIONS: GBV-C prevalence rates are lower in children with perinatal HIV infection than those reported for HIV-infected adults. With the exception of evidence that GBV-C viremic children had lower rates of Centers for Disease Control and Prevention HIV disease category C disease before GBV-C testing, we did not find evidence of improved HIV disease outcome in coinfected patients, but the number of HIV/GBV-C-coinfected children was small. PMID- 15876941 TI - Streptococcus pneumoniae and noncapsular Haemophilus influenzae nasal carriage and hand contamination in children: a comparison of two populations at risk of otitis media. AB - BACKGROUND: Australian Indigenous children living in remote areas have rates of tympanic membrane perforation as high as 60%, almost 100 times the prevalence in urban child care settings (<1%). Relative rates of pneumococcal nasal carriage do not reflect this difference in disease risk. METHODS: Cross-sectional comparison of nasal carriage and hand contamination in children younger than 4 years of age from urban child-care centers and Indigenous children 3-7 years of age from a remote community. Almost identical methods of nasal swab collection, transport and culture were used. Data on pneumococcal antimicrobial susceptibility patterns and serotypes are also reported. RESULTS: For Indigenous children compared with children in child care, the relative risk of nasal carriage of either pneumococcus or noncapsular Haemophilus influenzae was <2-fold [relative risk, 1.7; 95% confidence interval (CI), 1.5, 1.9], the risk of simultaneous nasal carriage was almost 3-fold (78% versus 28%; relative risk, 2.9; 95% CI 2.3, 3.5), and the risk of pneumococcal hand contamination was 8-fold higher (37% versus 4%; relative risk, 8.4; 95% CI 4.6, 15.2). For simultaneous hand contamination, the risk was 23-fold (8% versus 0.3%; relative risk, 23.1; 95% CI 2.9, 185.4). Remote Indigenous children also had a more diverse serotype distribution (25 versus 14 serotypes identified). CONCLUSIONS: Simultaneous nasal carriage of Streptococcus pneumoniae and H. influenzae and hand contamination are simple indicators of risk for use in studies of otitis media in populations at risk for tympanic membrane perforation. PMID- 15876942 TI - Kawasaki syndrome in Hawaii. AB - OBJECTIVE: To describe the incidence and epidemiology of Kawasaki syndrome (KS) in Hawaii. METHODS: Retrospective analysis of the State Inpatient Database for Hawaii residents hospitalized with KS during 1996 through 2001. RESULTS: During 1996 through 2001, 267 persons younger than 18 years of age living in Hawaii were hospitalized with KS; 226 (84.6%) were younger than 5 years of age. The average annual incidence for KS was 45.2 per 100,000 children younger than 5 years of age. The incidence was higher for children younger than 1 year of age than for those 1-4 years of age (74.3 and 37.5 per 100,000). The KS incidence for Asian and Pacific Islander children and for White children was 70.9 and 35.3 per 100,000, respectively. Incidence was highest among Japanese American children living in Hawaii (197.7 per 100,000). Honolulu County had the most KS patients (85.0%) and the highest incidence (53.1 per 100,000) among Hawaii counties. For children younger than 5 years of age hospitalized with KS, the median length of stay was 2 days, and the median hospital charge was $9379. CONCLUSION: During 1996 through 2001, the annual incidence rate for KS among children younger than 5 years of age in Hawaii was the highest in the United States. The incidence among Japanese American children in Hawaii was higher than that among other racial groups in the state and when compared with children living in Japan. PMID- 15876943 TI - Effectiveness of the Oka/GSK attenuated varicella vaccine for the prevention of chickenpox in clinical practice in Israel. AB - BACKGROUND: The varicella Oka/Merck vaccine has been shown to be very effective in clinical practice; however, several recent studies reported reduced effectiveness. The varicella Oka/GSK vaccine (Varilrix; GlaxoSmithKline Biologicals), which has been subjected to fewer effectiveness studies, was licensed in Israel for voluntary use in 2000. This study was planned to estimate the effectiveness of the varicella Oka/GSK vaccine among children 1-5 years of age. METHODS: Recent reports of varicella were retrieved from the Tel Aviv Health District. Two age-matched control subjects who attended the same day-care center and/or resided in the same neighborhood and had not contracted varicella in the past were recruited for each varicella case. The parents of case and control subjects were interviewed regarding the history of varicella vaccination and the severity of illness. RESULTS: A total of 151 case subjects (mean age, 2.9 +/-1.1 years) and 298 control subjects (mean age, 3.0 +/-1.1 years) were recruited. Vaccination rates for case and control subjects were 6.6% and 38.3%, respectively (P < 0.001). Disease among vaccinated children was significantly milder, as measured by several indices, including parental perception of the illness, presence of fever, time until the lesions dried and days of day care missed. Vaccine effectiveness against varicella of any severity with these figures was 88% (95% confidence interval, 77-94%), and effectiveness against moderate/severe illness was 100%. CONCLUSIONS: Our data demonstrate that the varicella Oka/GSK vaccine used in clinical practice is highly effective in prevention of varicella (especially moderate/severe disease) among children, including those attending day-care centers. PMID- 15876944 TI - Role of atypical bacteria and azithromycin therapy for children with recurrent respiratory tract infections. AB - BACKGROUND: The aim of this study of 352 patients, 1-14 years of age, with acute respiratory infections and a history of recurrent respiratory tract infections (RRTIs), and 208 healthy subjects was to evaluate whether Mycoplasma pneumoniae and Chlamydia pneumoniae played a role in causing acute respiratory episodes among children with RRTIs and whether specific antibiotic treatment for these bacteria could improve the acute episodes and reduce recurrences. METHODS: The patients were blindly randomized to receive azithromycin (10 mg/kg/d for 3 days weekly, for 3 weeks) together with symptom-specific agents or symptom-specific agents alone. Acute M. pneumoniae and/or C. pneumoniae infection was diagnosed if the child had a significant antibody response in paired sera and/or if the DNA of the bacteria was detected in nasopharyngeal aspirates. RESULTS: Atypical bacterial infections were identified for 190 patients (54.0%) and 8 healthy control subjects (3.8%; P < 0.0001). Short term (1-month) clinical success was significantly more frequent among the patients who had received azithromycin together with symptom-specific agents than among those who had received symptom specific agents alone, but the difference was significant only for the group of patients with atypical bacteria. In contrast, long term (6-month) clinical success was significantly more frequent among the patients who had received azithromycin in addition to symptom-specific agents, regardless of whether they experienced infections with atypical bacteria or other pathogens, although positive outcomes were significantly more frequent among those with atypical bacteria. CONCLUSIONS: Atypical bacteria seem to play a role among children with RRTIs, and prolonged azithromycin therapy can significantly improve the acute episodes and reduce the risk of recurrences. PMID- 15876945 TI - Diagnosis of catheter-related bloodstream infections among pediatric oncology patients lacking a peripheral culture, using differential time to detection. AB - BACKGROUND: Current methods for in situ diagnosis of catheter-related bloodstream infections require concurrent collection of central venous catheter (CVC) and peripheral vein (PV) blood cultures. Both the pain and inconvenience of PV cultures are undesirable. METHODS: A prospective study was conducted (August 2002 to March 2004) to assess the accuracy of diagnosing catheter-related bloodstream infections based on the difference in time to detection of blood cultures drawn concurrently from 2 lumens of a multilumen CVC. This difference in time to detection between 2 lumens was compared with results of the standard criterion with paired CVC and PV blood cultures. RESULTS: Twenty-one infectious episodes were categorized as catheter-related bloodstream infections and 38 as non catheter-related bloodstream infections. With a cutoff in difference in time to detection between 2 lumens of > or =180 minutes, the sensitivity of this test to diagnose a catheter-related bloodstream infection was 61% (95% confidence interval, 39-80%) and the specificity was 94% (95% confidence interval, 82-99%). In 4 of 7 episodes with false-negative results, the colony counts in cultures from both lumens were >400 colony-forming units/mL (maximal value reported), indicating the limitation of this method when both lumens of the catheter are colonized. With the pretest probability of catheter-related bloodstream infections ranging from 28% to 54%, the positive predictive value of a difference in time to detection between 2 lumens of > or =180 minutes for diagnosis of catheter-related bloodstream infections ranged from 81% to 93% and the negative predictive value ranged from 67% to 86%. CONCLUSION: Within the context of its limitations, this novel method provides an alternative for diagnosing catheter related bloodstream infections among patients with a CVC, without PV cultures. PMID- 15876946 TI - Malaria among refugee children at arrival in the United States. AB - BACKGROUND: Malaria infects >70% of children at any given time in highly disease endemic areas, such as parts of West Africa. Many infected children are asymptomatic. There are no published data regarding the prevalence of malaria among refugee children arriving in the United States from highly disease-endemic areas. This study was performed to determine the prevalence of malaria among asymptomatic children from West Africa. METHODS: A retrospective chart review for Liberian refugee children arriving in the United States was conducted from 1997 to 2000. RESULTS: Fifty-seven charts were reviewed. There were 28 (64%) positive thick and thin blood smears among the 44 children who had malaria smears performed during refugee medical screening examinations. There were 3 children who had been treated recently for malaria. An additional 3 children had splenomegaly and were treated for presumed malaria. Overall, the prevalence rate was 60% (34 of 57 cases). No sign or symptom, singly or in combination, was sufficient to predict the presence or absence of malaria. However, splenomegaly, fever and thrombocytopenia were highly specific for malaria and should alert clinicians to the likelihood of the diagnosis. Twenty-nine percent of children with malaria were asymptomatic. CONCLUSION: The results of this study suggest that refugee children arriving from West Africa have a significant burden of malaria. It is recommended that children who arrive in the United States as refugees from West Africa be subjected to a thorough physical examination, complete blood count and malaria blood smear. PMID- 15876947 TI - Outbreak of invasive Haemophilus influenzae serotype a disease. AB - Haemophilus influenzae serotype a is a rare cause of invasive disease. We report 5 cases of invasive H. influenzae type a that occurred in 3 infants living in a remote region of Alaska during the last 6 months of 2003. H. influenzae type a isolates from this outbreak were closely related as determined by pulsed field gel electrophoresis. Continued surveillance is necessary to monitor trends in H. influenzae invasive disease. PMID- 15876948 TI - Treatment of community-associated methicillin-resistant Staphylococcus aureus infections. PMID- 15876949 TI - Epidemic community-associated methicillin-resistant Staphylococcus aureus: modern times for an ancient pathogen. PMID- 15876950 TI - Selective IgA deficiency in children with recurrent parotitis of childhood. AB - Recurrent parotitis of childhood is defined as the relapsing form of juvenile (idiopathic) parotitis and represents a rare inflammatory disorder of the parotid gland with potentially significant morbidity. We reviewed the charts of patients who were diagnosed with inflammatory parotid diseases in our institution between 1992 and 2002. There were 91 patients presenting with juvenile parotitis (1 of 6117 of all clinical visits). Of these 91 cases, 23 patients (28%) had the relapsing form of juvenile parotitis, and the median number of episodes was 5 (range, 2-20). Laboratory investigations revealed that 5 patients had selective IgA deficiency. The prevalence (22%) is different from the cumulative prevalence of IgA deficiency in a healthy population (0.3%; P < 0.001). PMID- 15876951 TI - Antibodies against Haemophilus influenzae type b capsular polysaccharide and tetanus toxoid before and after a booster dose of the carrier protein nine years after primary vaccination with a protein conjugate vaccine. AB - IgG antibodies against Haemophilus influenzae type b (Hib) capsular polysaccharide (CPS) and tetanus toxoid (TT) were measured for 53 children, 10 years of age, before and 1 month after a booster dose of diphtheria-tetanus vaccine (DT). All children had been vaccinated at 3, 5 and 12 months of age with DT and a Hib-TT conjugate. Geometric mean concentrations of Hib CPS serum IgG antibody were 4.16 and 4.30 microg/mL before and after the DT booster, respectively. The geometric mean concentration of TT IgG antibody increased from 0.09 IU/mL to 4.58 IU/mL (P < 0.001). Hib CPS IgG levels remained well above protective titers for 9 years after 3 doses of Hib-TT appropriately spaced in infancy. A booster dose of TT did not affect Hib CPS antibody concentrations but induced a pronounced IgG response against TT. PMID- 15876952 TI - Amebic liver abscesses among neonates can mimic bacterial sepsis. AB - Amebic liver abscesses are unusual among neonates. Three (3.6%) of 83 children who presented to a university hospital with amebic liver abscesses in a 17-year period were neonates (<28 days of age). Patients presented with nonspecific clinical and laboratory findings mimicking fulminant neonatal sepsis. Diagnosis was delayed and was based on the presence of a hypoechoic lesion in the liver, antibodies against Entamoeba histolytica and microbiologic examination of pus. In addition to parenteral metronidazole treatment and early ultrasound-guided aspiration of the abscess, intensive care is vital for the treatment of neonates with amebic liver abscesses. PMID- 15876953 TI - Severe acute tonsillitis caused by Rothia dentocariosa in a healthy child. AB - A 4-year-old Japanese girl developed a sore throat and high fever. Her tonsils were enlarged, red and covered with a thick white membrane. There was marked leukocytosis (26,600 leukocytes per mm) and elevated C-reactive protein levels (23.3 mg/dL). Rothia dentocariosa was recovered from the throat swab; many Gram positive cocci were observed in the smear from the pseudomembrane covering the tonsil. PMID- 15876954 TI - Cytochrome c and tumor necrosis factor-alpha values in serum and cerebrospinal fluid of patients with influenza-associated encephalopathy. AB - Cytochrome c and tumor necrosis factor-alpha concentrations were measured in serum and cerebrospinal fluid samples from 10 patients with influenza-associated encephalopathy. In the acute exacerbation phase, serum tumor necrosis factor alpha and cytochrome c values were high in patients with a poor prognosis. In the convalescent phase, cerebrospinal fluid cytochrome c values increased remarkably in patients with subsequent brain atrophy. PMID- 15876955 TI - Right atrial mass in a child with disseminated coccidioidomycosis. AB - A 10-year-old patient with known coccidioidomycosis relapsed and had dysrrhythmias and a right atrial mass. Histopathology and culture after surgical removal revealed that this was a vegetative mass infected with Coccidioides spp. We believe that this is the first case of coccidioidal endocarditis to be reported. PMID- 15876956 TI - Multifocal bone marrow involvement in cat-scratch disease. AB - We describe 2 children with prolonged fever of unknown origin and prominent skeletal pain who had multifocal bone disease caused by Bartonella infection. Initial radiologic studies, including plain films, radionuclide scintigraphy and computed tomography, yielded negative results. In both cases, magnetic resonance imaging revealed multiple enhancing bone marrow lesions consistent with clinical symptoms. Microbiologic diagnoses were established serologically. PMID- 15876957 TI - Paraspinal mass in a seven-year-old girl. PMID- 15876958 TI - Varicella zoster virus encephalitis in a previously healthy five-year-old child with herpes zoster ophthalmicus. PMID- 15876959 TI - Seroprevalence of human herpesvirus 8 in children in Seoul, Korea. PMID- 15876960 TI - Oral lesions in a group of kidney transplant patients. AB - AIM: To assess the prevalence of oral mucosa lesions (OL) in a group of kidney transplant (KT) patients, and analyze possible OL associations with one another and with drugs use and relevant clinical and laboratory variables. METHODOLOGY: Transversal study, in which consecutive KT patients from the nephrology outpatient service at a General Zonal Hospital were examined. The prevalence of several types of OL was assessed, and their possible statistical associations with one another and total leukocyte count, renal function, the simplified oral hygiene index (S-OHI), cyclosporin-A (CsA) dose and blood levels, and nifedipine use was analyzed. A logistic regression model was used to analyze the association between gingival hyperplasia (GH) and CsA dose and nifedipine use. RESULTS: Fifty (55.6%) men and forty (44.4%) women were studied. Sixty percent of the patients had at least one OL. Oral candidiasis (OC) was found in 18.7%; 13% had lesions clinically compatible with hairy leukoplakia (CHL). An association was found between OC and CHL (P<0.05). Saburral tongue (ST) was found in 22% of the patients and gingival hyperplasia (GH) in 49%, which was distributed as follows: Grade 1 in 11 (12.2%); Grade 2 in 26 (28.9%), and Grade 3 in 7 (7.8%). Logistic regression results showed an association between GH and poor oral hygiene (P< 0.001), but not to either CsA dose or blood levels, or nifedipine use (P=0.075). CONCLUSION: Sixty percent of the KT patients had at least one OL. The association between GH and poor oral hygiene corroborate the need for oral hygiene practices supervision in the transplant patient. PMID- 15876961 TI - Expression of major histocompatibility complex class II and costimulatory molecules in oral carcinomas in vitro. AB - Recognition in the 1980 s that keratinocytes can express class II molecules of the Major Histocompatibility Complex (MHC) first raised the possibility that these cells might have an immunological function, and may even act as antigen presenting cells (APC). For effective T lymphocyte activation, APC require, in addition to MHC II, appropriate costimulatory signals. The aim of this study was to determine the expression of MHC class II and the co-stimulatory molecules CD40, CD80 and CD86 in keratinocytes derived from healthy oral mucosa and oral carcinomas. Using flow cytometry, it was confirmed that oral keratinocytes, switch on, expression of MHC class II molecules after stimulation with IFNgamma in vitro. All keratinocyte lines expressed CD40 constitutively; by contrast, CD80 and CD86 were universally absent. Loss of CD80 and CD86 may be one means whereby tumours escape immunological surveillance. PMID- 15876962 TI - Ameloblastoma. Diagnosis by means of FNAB. Report of two cases. AB - INTRODUCTION: Ameloblastomas are the most frequent odontogenic tumors of the maxilla. In spite of their benign cytohistological appearance, they behave as invasive recurring tumors, with the possibility of metastasis. FNAB is a rapid, bloodless test that provides a pre-surgical diagnosis, thus, on occasions avoiding the need for diagnostic biopsies. We present the cytological characteristics of two cases of jugal recurrences of mandibular ameloblastomas diagnosed by FNAB, as well as their cytohistological correlation. CLINICAL CASES: Two patients, a 36-year-old woman, and a 62-year-old male who both attended with mandibular swelling of a few months evolution. In both cases the first diagnostic approximation was the histological study of the tumoral mass, together with the radiological studies. Following therapeutic extirpation both cases recurred. The diagnosis of the recurrences was established cytologically by means of FNAB. The cytologic smears revealed a granular background with isolated macrophages and giant multinucleate cells and an abundant epithelial cellularity of basaloid appearance arranged in cohesive groups forming images of peripheral palidasing, as well as small groups of squamous metaplastic cells. DISCUSSION: FNAB is considered to be a rapid, bloodless and reliable method for the diagnosis of ameloblastoma. The cytology of these tumors reveals components of the lesion that, in general, are sufficient for the diagnosis of ameloblastoma, especially in cases of recurrence. PMID- 15876963 TI - Jacob's disease associated with temporomandibular joint dysfunction: a case report. AB - Jacob's disease is regarded a rare condition in which a joint formation is established between an enlarged mandibular coronoid process and the inner aspect of the zygomatic body. Chronic temporomandibular joint (TMJ) disk displacement has been proposed as etiological factor of coronoid process enlargement. We present a 23-year-old woman with long-standing TMJ dysfunction and restricted interincisal opening, who developed a progressive zygomatic asymmetry. The patient underwent treatment by intraoral coronoidectomy and homolateral TMJ arthroscopy in the same surgery. The histopathological diagnosis of the coronoid sample was cartilage-capped exostoses with presence of articular fibrous cartilage. Although the low prevalence of this entity, it should be considered as a possible diagnosis in patients with progressive limitation of mouth opening, although a TMJ syndrome may be present as a cause of this entity. PMID- 15876964 TI - Heart and periodontal diseases: does evidence exist of association? AB - Cardiovascular diseases represent one of the main causes of death in developed societies. These diseases are not only important because of the mortality they generate, but also due to the high number of affected persons under treatment that the health systems of different countries have to support. Hence, the early detection of risk factors triggering these diseases has been seen as the most effective mechanism to prevent them. Over the last few years periodontal pathology has been more or less related to the suffering of cardiovascular diseases. This fact could be of great clinical and epidemiological relevance owing to the actual considerable prevalence of periodontal diseases. In the present review, the clinical and physiopathological factors which scientifically support this association are analyzed,reaching the conclusion that further studies are needed to have the proof that said association represents a determinate risk factor for the suffering of cardiovascular diseases. PMID- 15876965 TI - Day case oral surgery in pediatric patients during the year 2000 in the University of Barcelona Dental Clinic (Spain). AB - AIM: A study is made to show that most pediatric oral surgical interventions can be performed on a day case surgery or ambulatory basis, and to describe the activity of the Master of Oral Surgery and Implantology (Barcelona University Dental School, Spain) in the year 2000. DESIGN: The study documents the surgical activity of the Units of Oral Surgery and Laser Surgery in pediatric patients, in the context of the Master of Oral Surgery and Implantology (Barcelona University Dental School, Spain). RESULTS: Of the 3187 operations carried out in our Service in this period, 489 involved patients under 18 years of age. The surgical removal of third molars was the most common intervention (55.6%), followed by other surgical or non-surgical tooth extractions (33.6%) and other interventions (10.6%) comprising fenestration of impacted canines or other teeth, frenectomies, the removal of mucoceles, and tooth relocations. Complications after surgical third molar extraction were recorded in 15.93% of cases, the most frequent problem being the pain and swelling (4.35% each) characteristic of postoperative inflammation. CONCLUSIONS: Our experience shows that day case oral surgery can be safely performed in pediatric patients, provided the required means and health care personnel resources are available. PMID- 15876966 TI - Mandibular ameloblastoma. A review of the literature and presentation of six cases. AB - Ameloblastoma is a benign odontogenic tumour of epithelial origin without induction in the connective tissue. In treating this type of tumour, it is important to assess the clinical type (solid, multicystic, unicystic, peripheral), localisation, and size of the tumour as well as age of the patient. Articles have recently been published with the purpose of providing updated knowledge and therapeutic approaches to ameloblastoma. We present six cases of patients with localised mandibular ameloblastoma who were treated during the last seven years. We present data on clinical appearance, histological characteristics, and therapeutic approach that was used, which included excision of the lesion, perilesional drilling of the bone, or block resection, according to the type of ameloblastoma. Lastly, we analysed follow-up measures and the rate of recurrence in these patients. The aim of this paper is to review the concepts relating to ameloblastoma that have been published recently and to assess their influence on the clinical attitude taken when facing this pathology, using our experience with ameloblastoma as a starting point to illustrate this discussion. PMID- 15876967 TI - Giant tonsillolith: report of a case. AB - Tonsilloliths or tonsil stones are calcifications that form in the crypts of the palatal tonsils. These calculi are composed of calcium salts either alone or in combination with other mineral salts, and are usually of small size - though there have been occasional reports of large tonsilloliths or calculi in peritonsillar locations. We present the case of a 55-year-old woman with a one year history of dysphagia and pharyngeal discomfort with a foreign body sensation, though the manifestations had recently intensified. Exploration of the oral cavity revealed a hard bulging submucosal mass in the region of the soft palate, at right anterior tonsillar pillar level. The mucosa overlying the lesion appeared erythematous. Computed tomography revealed a large, delimited and highly calcified oval image measuring 2.5 x 1.5 cm, which was subsequently surgically removed. PMID- 15876968 TI - Calcifying odontogenic cyst associated with complex odontoma: case report and review of the literature. AB - We report a calcifying odontogenic cyst associated with odontoma (COCaO) and an included permanent canine in the superior maxilla, in a 19 year-old-man. The calcifying odontogenic cyst (COC) was first described as a distinct entity by Gorlin et al in 1962. The lesion is a mixed odontogenic benign tumor, and although most of the cases present cystic characteristics, a few are of the solid type (15%), and its rare malignant transformation is well documented. The COC may occur in association with other odontogenic tumors, the most common is the odontoma, occurring in about 24% of the cases. For this association the term Odontocalcifying odontogenic cyst has been suggested. Radiographically is a well defined mixed lesion and histologically consists of a large cyst. In the central area of the cyst enamel and dentin deposits can be found, irregularly distributed in areas and in other parts it takes on a well defined organoid aspect. A thorough review of literature takes place and the pathogenesis is discussed. PMID- 15876969 TI - Intraosseous odontoma erupted into the oral cavity: an unusual pathology. AB - OBJECTIVES: Odontoma arising in the extraosseous soft tissue is extremely uncommon. Two forms of odontoma are presently recognized: peripheral odontoma and erupted odontoma. We report an erupted odontoma arising in the posterior maxilla, and discuss the main differences between both forms of clinical presentation. STUDY DESIGN: Case study. RESULTS: A 23-year-old man presented with a firm gingival mass on the left posterior maxilla, that had gradually enlarged over one year. Orthopantomography showed a dense radiopaque mass occupying the posterior portion of the left maxilla. The pathology was reported as complex odontoma. CONCLUSIONS: This is the eleventh reported case of erupted odontoma. Unlike peripheral odontoma, erupted odontoma is generally a complex odontoma, and affects older patients. PMID- 15876970 TI - Metastatic tumours in upper maxillary bone of esophageal adenocarcinoma. A case report. AB - Metastases in the oral cavity are rare lesions which represent approximately 1% of all malignant neoplasms in the oral cavity. Oral metastases are located in the mandible 80-90% on average, the maxilla location being rarer. Metastases in mouth soft tissue are also rare, and within these it is on the gums where they more frequently occur. Primary tumours which metastasize to mouth are most commonly: lung, breast and kidney. Oral cavity metastases appear as a result of distant disease spreading and show wrong prognosis, with short survival. Here we present a clinical case of a patient diagnosed with esophagus adenocarcinoma which presented metastasis in upper-left maxillary bone. PMID- 15876971 TI - Pain and inflammation in 41 patients following the placement of 131 dental implants. AB - AIMS: To study pain and swelling in the first week after dental implant placement. MATERIAL AND METHODS: A total of 131 implants were placed in 41 patients (17 males and 24 females) under local anesthesia. Pain was scored by means of a verbal and visual analog scale (VAS), while swelling was evaluated by a verbal scale. RESULTS: Most patients who experienced pain reported the latter to be slight, with a peak intensity 6 hours after the operation in 41.5% of cases. A significant relation was observed between pain and the number of implants placed. Swelling was moderate in most patients who reported pain -- peak intensity of inflammation being recorded after 48 hours in 48.8% of cases. A significant association was observed between swelling and older patients, the placement of more than four dental implants, and operations in which sinus lift or bone regeneration procedures were carried out. Swelling was greater in patients with implants positioned in the posterior sector versus those placed in the anterior zone, and in those located at free extreme or totally edentulous patients, versus those placed in interdental spaces. CONCLUSIONS: Following implant placement, pain tends to be mild, with moderate inflammation. The latter is seen to be greater in older patients, in subjects receiving a larger number of implants, and in interventions involving regenerative techniques. PMID- 15876972 TI - Melanoma of the oral mucosa. Clinical cases and review of the literature. AB - The appearance of primary melanomas of the oral mucosa is uncommon. The aggressiveness of this entity and the absence of any standardized treatment protocol make the prognostic unfortunate. The difficulty to obtain free surgical margins, the elevated tendency to invade in depth and the early haematogenous metastasis have been referred as features which may explain its bad prognosis, even in comparison with cutaneous melanoma. However, no large clinical series exist and actually, clinical cases are the main source of information. Due to the absence of any treatment modality which may substantially increase long-term survival, we suggest the use of resective surgery with wide margins and early diagnosis by means of biopsy for suspicious melanotic-pigmented lesions. In this work we present 2 new cases of primary melanoma of the oral mucosa, with a follow up period of 72 and 12 months respectively, and we make a review of the literature in relation with this rare entity. PMID- 15876973 TI - Synovial chondromatosis of the temporomandibular joint: a clinical, radiological and histological study. AB - Synovial chondromatosis (SC) is a cartilaginous metaplasy of the mesenchymal remnants of the synovial tissue of the joints. It is characterized by the formation of cartilaginous nodules in the synovium and inside the articular space (loose bodies). SC mainly affects to big synovial joints such as the knee and the elbow, being uncommon the onset within the TMJ, where 75 cases have been published. The main symptoms are pain, inflammation, limitation of the movements of the jaw and crepitation. Different methods of diagnosis include panoramic radiograph, CT, MR and arthroscopy of the TMJ. We report a new case of unilateral SC of the TMJ, including diagnostic images, treatment performed and histological analysis. PMID- 15876974 TI - Bifid condyle: case report. AB - The double headed mandible condyle is a rare alteration that is frequently diagnosticated as an incidental finding in a panoramic radiograph. The different theories about its etiology are explained in this article. Symptoms described with bifid condyles vary from case to case, but in most instances are absent. Computer tomography is the ideal imaging method to evaluate the condyle morphology and to role out any degenerative process to achieve the differential diagnosis. Treatment is conservative for symptomatic patients with TMJ disorders, surgical treatment is described in literature in TMJ ankylosis that develops in bifid condyles secondary to trauma. PMID- 15876975 TI - Spindle cell carcinoma. PMID- 15876976 TI - Ameloblastoma. PMID- 15876977 TI - Vascular dementia. Advances in nosology, diagnosis, treatment and prevention. AB - Ischemic or hemorrhagic cerebrovascular disease (CVD) produces injury of brain regions important for executive function, behavior, and memory leading to decline in cognitive functions and vascular dementia (VaD). Cardiovascular disease may cause VaD from hypoperfusion of susceptible brain areas. CVD may worsen degenerative dementias such as Alzheimer disease (AD). Currently, the global diagnostic category for cognitive impairment of vascular origin is vascular cognitive disorder (VCD). VCD ranges from vascular cognitive impairment (VCI) to VaD. The term VCI is limited to cases of cognitive impairment of vascular etiology, without dementia; VCI is equivalent to vascular mild cognitive impairment (MCI). Risk factors for VaD include age, hypertension, diabetes, smoking, cardiovascular disease (coronary heart disease, congestive heart failure, peripheral vascular disease), atrial fibrillation, left ventricular hypertrophy, hyperhomocysteinemia, orthostatic hypotension, cardiac arrhythmias, hyperfibrinogenemia, sleep apnea, infection, and high C-reactive protein. Research on biomarkers revealed increased CSF-NFL levels in VaD, whereas CSF-tau was normal. CSF-TNF-alpha, VEGF, and TGF-beta were increased in both AD and VaD. VaD shows low CSF acetylcholinesterase levels. This condition responds to acetylcholinesterase inhibitors, confirming the central role of cholinergic deficit in its pathogenesis. Evidence strongly suggests that control of vascular risk factors, in particular hypertension, could prevent VaD. PMID- 15876978 TI - Pathology and pathophysiology of vascular cognitive impairment. A critical update. AB - The prevalence, morphology, and pathogenesis of vascular dementia (VaD), recently termed vascular cognitive impairment (VCI), and of mixed dementia (Alzheimer disease associated with vascular encephalopathy) are a matter of discussion and clinical diagnostic criteria for these disorders slow low sensitivity and variable specificity. In Western memory clinic-based series, VaD/VCI is suggested in 8-10% of cognitively impaired elderly subjects. Its prevalence in autopsy series ranges from 0.03% to 58% with reasonable values of 4-10%. It has been related to focal, multifocal or diffuse cortical and/or subcortical microinfarcts and lacunes often affecting strategically important brain areas (thalamus, frontobasal and/or limbic systems), hemispheric white matter lesions and, less often, large brain infarcts. They result from systemic, cardiac or local large or small vessel disease. The pathogenesis of cerebrovascular lesions is multifactorial and their pathophysiology affects neuronal systems involved in cognition, memory, and behavior. Vascular pathology often coexists with Alzheimer disease (AD) and other dementing disorders; 25-60% of dementia cases show both AD and vascular lesions. The lesion patterns in ''pure'' VCI with predominant multiple small (subcortical) lesions related to arteriosclerosis and microangiopathies and in mixed dementia (AD associated with vascular encephalopathy), more often showing large infarcts, suggest different pathogenesis of both types of lesions. Due to the high variability of cerebrovascular pathology and its causative factors no validated neuropathological criteria exist for VCI and mixed dementia. In very old subjects selective hippocampal sclerosis may be associated with multiple other vascular pathologies. Minor cerebrovascular lesions, except for severe amyloid angiopathy, appear not essential for cognitive decline in full-blown AD, while both mild AD type pathology and small vessel disease may interact synergistically in ''unmasking'' or promoting dementia. AD pathology is significantly less severe in the presence of cerebrovascular lesions. Further studies are needed to validate diagnostic criteria for VaD/VCI and to clarify the impact of vascular lesions on cognitive impairment as a basis for more precise clinical diagnosis, early prevention and management. PMID- 15876979 TI - Blood pressure and dementia. AB - Hypertension has proven to be a major predisposing factor for the development of both vascular and degenerative (Alzheimer's) dementias, either following stroke or gradually through more insidious microcerebrovascular processes. In the latter case the interval between the respective manifestations of hypertension and dementia may vary between a few years and several decades. The temporal relationships may become complicated by the finding that blood pressure tends to fall shortly before the onset of overt Alzheimers' disease. Whether or not timely antihypertensive regimens may delay or even prevent the development of dementias later in life is still an ''educated'' guess, as long there has been no comprehensive trial comparing the potential of the different antihypertensive drug classes in this regard. Until then, the class of dihydropyridine calcium antagonists (exemplified by nitrendipine in the Syst-Eur trial) is the only category having statistically been proven to be of substantial value for prevention of Alzheimer's disease. PMID- 15876980 TI - Cerebrovascular disease and the pathophysiology of Alzheimer's disease. Implications for therapy. AB - Dementia is a disease of the elderly, and although there are many causes of dementia, Alzheimer's disease (AD) and vascular dementia (VaD) account for the majority of cases world- wide. Many patients with dementia have radiological and neuropathological features of AD and VaD, with the classical neurofibrillary tangles and senile amyloid-beta (Abeta) plaques of AD together with the cerebral infarcts of VaD. In this review we examine the close relationship between AD and VaD and suggest that the age changes in cerebral blood vessels that are the basis of cerebrovascular disease and VaD may also be responsible for the failure of elimination of Abeta from the brain in AD. Abeta appears to be eliminated along the perivascular pathways by which interstitial fluid (ISF) drains from the brain (effectively the lymphatics of the brain). In aged individuals, insoluble Abeta amyloid fibrils are deposited in the ISF drainage pathways resulting in cerebral amyloid angiopathy (CAA). We review the evidence that age changes in cerebral arteries and cerebrovascular disease inhibit the perivascular drainage of ISF and Abeta along the walls of cerebral arteries resulting in the accumulation of insoluble and soluble Abeta in the brain in AD. Therapies for AD are reviewed, especially those involving immunotherapy for the removal of insoluble Abeta from the cerebral cortex and the facilitation of drainage of ISF and soluble Abeta from the brain. PMID- 15876981 TI - Cerebral amyloid angiopathy and dementia. AB - Cerebral amyloid angiopathy (CAA) is a fundamental part of the pathology of many disorders causing dementia and/or cerebral haemorrhage. In Alzheimer's disease (AD), CAA is due to the deposition of amyloid alpha protein (Abeta) within the adventitia and media of leptomeningeal and brain parenchymal arteries. Although virtually all cases of AD show CAA to a greater or lesser extent, the brain distribution of CAA is not uniform with the occipital lobe being the most commonly and most severely affected region. In vessels affected by CAA, local muscle and elastic elements are lost and replaced by amyloid fibrils, thereby weakening the overall structure of the vessel. Consequently, CAA predisposes towards cerebral infarction and cerebral haemorrhage, though the clinical affects of CAA in AD are mostly silent, or at least are ''masked'' by the greater degree of neuronal dysfunction induced by senile plaque (SP) formation and neurofibrillary degeneration. Nonetheless, major cerebral infarctions with focal neurological deficits can occur in some cases of AD, and CAA is a major cause of fatal intracerebral (lobar) haemorrhage. CAA may also contribute to white matter lesions (myelin loss) in AD by inducing ischaemia through autoregulatory dysfunction. Although the Abeta protein deposited within blood vessels in AD is similar in chemical composition to that deposited in the brain parenchyma in SP, there is no clear relationship between the 2 pathologies. Indeed, when CAA is high, SP formation may be low, and vice versa. As if to emphasise these differences, Abeta within CAA is mostly Abeta40 whereas that within SP is Abeta42. Such compositional differences may reflect differences in source, with Abeta in SP being derived from nerve cells and Abeta in CAA having a local vascular origin. Although certain inherited forms of CAA with cerebral haemorrhage are associated with autosomal dominant mutations in APP and other genes (cystatin-C, transthyretin, gelsolin, ABrit, ADan), in most cases of AD CAA does not associate clearly with any genetic risk factor other than APO E beta4 allele, which appears to increase the severity of CAA in a dose dependent manner, especially within the occipital cortex. Genotype/phenotype correlations may be helpful in understanding the development of CAA in AD and other disorders. Why blood vessels in the occipital lobe should be most susceptible to CAA in AD remains unclear, though this pattern of blood vessel involvement does not seem to be recapitulated in other disorders in which CAA is the principal pathological change. PMID- 15876982 TI - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an adult-onset hereditary syndrome characterized by recurrent TIAs and strokes, cognitive decline and dementia, migraine with aura (+/-40% of patients), and psychiatric disturbances (+/-30% of patients). Affected individuals have prominent signal abnormalities on brain MRI. Symmetrical white matter abnormalities are invariably seen and often small subcortical infarcts are also present. The extent of the MRI lesions increases with age, from subtle white matter abnormalities in the anterior temporal poles in the early 20 years to confluent white matter lesions with subcortical infarcts and microbleeds in the 6(th) decade. A typical arteriopathy with electron dense granular depositions in the media of small cerebral arteries underlies this disorder. These arterial lesions can be found, to a lesser extent, in extra-cerebral arteries such as skin arterioles. In 1996, the defective gene in CADASIL was discovered to be NOTCH3. NOTCH3 encodes a 300-kd transmembrane protein with a receptor and cell signal transduction function. Mutations are almost always missense mutations causing the loss or gain of a cysteine residue and are detected in over 90% of patients. How alterations in NOTCH3 lead to the CADASIL phenotype has yet to be elucidated. PMID- 15876983 TI - Aneurysms of the ophthalmic (C6) segment of the internal carotid artery: treatment options and strategies based on a clinical series. AB - AIM: Treatment of ophthalmic segment aneurysms is technically demanding and still associated with a relatively high morbidity and mortality. The refinements of surgical techniques combined with the development of indirect methods of treatment have greatly improved the outcome in treating these lesions. We present our clinical experience and discuss treatment strategies. METHODS: Seventy-eight consecutive patients with 88 ophthalmic segment aneurysms were admitted to our service from January 1997 to June 2003. Forty-three patients presented with unruptured aneurysms and 35 presented with subarachnoid hemorrhage (SAH). Management strategies included surgical clipping alone in 53 patients, clipping and hemicraniectomy in 2, coiling in 17, external carotid artery-middle cerebral artery (ECA/MCA) by-pass in 2, and coil occlusion of the internal carotid artery in 2. Two patients underwent no treatment. RESULTS: In the group of 41 treated patients with unruptured aneurysms, 40 (97.6%) had good outcomes (GOS 1-2) and 1 patient had poor (GOS 3) outcome at discharge. Procedure-related morbidity was 15.7% (8/51 procedures), and permanent morbidity was 9.75% (4/41 patients). In the 35 patients who presented with SAH, mortality was 14.3% (5 patients); at discharge, 21 patients (60%) had good (GOS 1-2) and 9 (25.7%) poor (GOS 3) outcomes. The overall outcome was good (GOS 1-2) in 63 patients (80.8%) and poor (GOS 3-4) in 10 patients (12.8%). Overall mortality was 6.4% (5 patients all with SAH). CONCLUSIONS: Direct obliteration of the aneurysm utilizing advanced surgical techniques is our preferred treatment approach, whenever possible. In case of unclippable large or giant aneurysms, the surgical or endovascular occlusion of the proximal internal carotid artery with or without an extracranial intracranial by-pass is an option. A highly skilled team including a cerebrovascular and an endovascular surgeon is essential to achieve good outcomes in treating these lesions. PMID- 15876984 TI - Use of chronic sacral nerve stimulation in neurological voiding disorders. AB - AIM: Neurogenic low urinary tract dysfunctions unresponsive to medical and conservative therapy are difficult to manage. Nowadays they can be treated with Sacral Nerve Stimulation (SNS), even if clinical experiences reported in literature are still limited. METHODS: We performed SNS in 6 patients with neurogenic bladder: 3 patients had incontinence-urgency (1 myelitis, 1 multiple sclerosis, 1 autonomic polineuropathy) and 3 patients had urinary retention (1 incomplete spinal cord lesion, 1 operation for discal hernia T5-T6, 1 hysterectomy). RESULTS: Among cases with incontinence-urgency we achieved complete control of the bladder in 2 patients while in 1 patient the number of urinary losses was reduced of the 80%. In 2 patients with urinary retention we obtained complete recovery of the bladder function, while in 1 patient the number of cateterisms/die reduced of 50%, the urinary volume for micturion increased and residual urinary volume decreased. Results were unchanged during the follow-up (maximum 26 months), except for 1 patient in which a partial loss of effectiveness occurred. CONCLUSIONS: Chronic electric stimulation of S3 sacral roots via an implanted neuroprotesis is therefore an effectiveness, save and promising therapeutic option in treatment of neurogenic bladder dysfunctions. PMID- 15876985 TI - The paraclinoid aneurysms and the distal dural ring: a new classification. AB - AIM: The aneurysms of the paraclinoid region are a heterogeneous group of lesions, whose terminology and classification are contradictory because of the complex anatomy and the lack of key anatomical references. A surgical anatomical study to identify an anatomical reference is presented, a new classification of these aneurysms is proposed, and radiological guidelines are given to distinguish an intra from extradural lesion on the x-rays. METHODS: A surgical anatomical study of the paraclinoid region has been performed on 10 human cadaveric heads, formalin-fixed and injected with colored silicone rubber, and on skulls, with particular emphasis on the distal dural ring (DDR), anterior clinoid process (ACP) and their relationships with the cavernous, clinoid and ophthalmic segments of the internal carotid artery. On the basis of anatomical considerations a new classification of the aneurysms of the paraclinoid region centered on the DDR has been presented. Furthermore, 50 skulls and 5 injected cadaveric heads underwent radiological evaluation in order to develop x-ray criteria to identify the DDR. RESULTS: The key anatomical element is the DDR. This allows the differentiation of aneurysms of the paraclinoid region into 4 groups (supra-DDR, para-DDR, infra DDR, transitional). The radiological identification of the planum sphenoidale, tuberculum sellae and optic canal allows to distinguish intra versus extradural lesions. CONCLUSIONS: A new classification of the aneurysms of the paraclinoid region which uses the DDR as key anatomical element is proposed. Radiological guidelines able to identify the DDR and, thus, to distinguish intra versus extradural aneurysms are provided. PMID- 15876986 TI - Pharmacological complications of the chronic baclofen infusion in the severe spinal spasticity. Personal experience and review of the literature. AB - AIM: To evaluate the pharmacological complications and side effects of the chronic intrathecal baclofen infusion in patients suffering from intractable spinal spasticity in our Department as well as in the literature. METHODS: The pharmacological complications and side effects observed at long-term follow-up in 25 patients with chronic intrathecal baclofen infusion are evaluated; the search of the literature complications was carried out in Medline and in Ingenta web sites. RESULTS: We have been able to observe in our series 3 (12%) patients with hypotonia, 2 with impairment on erection (8%) and 1 (4%) with constipation; 5 (20%) patients showed also tolerance but only 1 (4%) needed a ''drug holiday''. In the literature the side effects range from 4% to 16%. Moreover the tolerance is reported from 3% to 15%. The overdose has been reported from 0% to 14%, while the syndrome of withdrawal is reported in 16 patients with 6 fatalities. In our series these two last complications were not observed. CONCLUSIONS: Lethal pharmacological complications are rare and, if promptly diagnosed and treated, are reversible. Side effects of minor importance can impair the patient quality of live and needed to be adjusted to patient's requirements. This kind of therapy should be restricted to specialized centres. PMID- 15876987 TI - [Laparoscopic treatment for Crohn's disease]. AB - AIM: Several studies have demonstrated the feasibility and safety of laparoscopic surgery for Crohn's disease. A trend towards less morbidity as compared to laparotomy has been suggested. However, problems noted early in the experience may have prevented the optimal benefit from having been conferred. Accordingly, the aim of this study was to evaluate perioperatively those patients. METHODS: All 51 patients with Crohn's disease who underwent an intestinal resection at Cleveland Clinic Florida between January 1997 and December 1998 were analyzed. RESULTS: Seventeen patients underwent laparoscopic treatment: there were no significant differences between the 2 groups as to age, gender, incidences of comorbidity, prior laparotomy, or the use of anti-inflammatory and immunosuppressive agents. Similarly, there were no significant differences between the 2 groups as to either surgical indication, intraoperative findings, or procedure performed. Moreover, there were no significant differences concerning total anesthetic time or surgical operative time, the incidence of intraoperative morbidity or need for enterolysis or stoma construction, use of intraoperative endoscopy, or need for transfusion. Significant differences were noted in the duration of patient controlled analgesic usage (3.1 days in the laparoscopic group vs 3.9 days, respectively; p = 0.03), the incidence of postoperative morbidity (7/17 patients in the laparoscopic group vs 27/34 patients: p = 0.01), and length of hospital stay (6.4 days in the laparoscopic group vs 9.6 days, respectively; p = 0.05). CONCLUSIONS: In this retrospective cohort comparative study, laparoscopic intestinal resection for Crohn's disease, when compared to laparotomy, was associated with a short duration of patient controlled analgesic usage, a lower incidence of postoperative morbidity and a shorter hospital stay, without significantly increased operative time. PMID- 15876988 TI - [Early vs conservative treatment of acute cholecystitis. Personal experience and review of the literature]. AB - AIM: The aim of this retrospective study was to compare the results of early cholecystectomy and conservative treatment in acute cholecystitis. METHODS: From January 1998 to December 2002, 134 patients were admitted to our Department with the diagnosis of acute cholecystitis. Eighty-nine patients (66%, Group 1) were cured with conservative treatment (i.e. fast, broad-spectrum antibiotics, fluid and analgesic drugs), 45 patients (34%, Group 2) were submitted to early cholecystectomy. The 2 groups were matched for age, sex, laboratory results and echographic findings. RESULTS: The morbidity was 32.5% in Group 1 versus 15.5% in Group 2 (p < 0.05). Mean hospital stay was 18 days in Group 1 as compared to 10.5 days in Group 2 (p < 0.05). At follow-up (12 months) the mean period before starting again a normal working and social life was 57 days in Group 1 and 33 days in Group 2. CONCLUSIONS: The results of this study showed that early cholecystectomy was the gold standard in the treatment of acute cholecystitis, since it reduces morbidity, hospital stay and absence from working and social life in a statistically important way. PMID- 15876989 TI - [Non-recurrent inferior laryngeal nerve: anatomy, frequency and surgical considerations]. AB - AIM: Systematic exposure and isolation of the inferior laryngeal nerve in thyroid surgery is a secure method to prevent complications. The knowledge of the nerve anatomy and its anomalies is essential to reduce laryngeal nerve injuries. By a re-examination of literature we underline the importance and some particular anatomical aspects of the anomalies which, even if rare, present an incidence of about 1%. METHODS: From January 1995 to December 2002 in our hospital we performed 299 interventions for thyroid diseases: total thyroidectomy (254 cases), partial thyroidectomy (45 cases). In 2 cases we observed a high variant (type I) of non recurrent laryngeal nerve. RESULTS: The global incidence of non recurrent laryngeal nerve was 0.66% (2/299 operations) and 0.72% if we consider the explorations of the cervical right side (2/274) where this anomaly is more frequent. The incidence of nerve injuries was 2/299 (0,66%); we had no complications in the 2 cases of non recurrent nerve observed. CONCLUSIONS: Our experience in thyroid surgery confirms the common opinion that only the knowledge of the laryngeal nerve anatomy and the awareness of its anomalies, associated to the systematic exposure of the nerve during surgery can prevent surgical injuries. PMID- 15876990 TI - Management of cervical aerodigestive injuries. AB - The management of patients with cervical injuries is highly controversial. Some authorities advocate mandatory exploration for all such injuries, while others advocate selective exploration. This paper will objectively review the evidence supporting each approach. The non-operative approach may be pursued through a variety of diagnostic modalities and this paper will also review the evidence supporting their use in cervical trauma. A clear understanding of these modalities and their relative merits is mandated by the potential severity of cervical injuries and their need for rapid intervention. PMID- 15876991 TI - [Clinical role of interleukin-2 in the surgical treatment of liver metastasis due to colon adenocarcinoma]. AB - The surgical treatment of liver metastasis due to colorectal cancer can substantially modify the natural history of the disease, mainly when it is associated with effective medical treatment. Chemotherapy, via systemic or locoregional (intrahepatic) administration, has 2 possible objectives: as adjuvant treatment, to prevent or delay disease recurrence; as neo-adjuvant treatment, mainly interesting for the surgeon, to allow resective surgery in responding patients previously considered not-operable. Unfortunately, the severe immune deficiency associated with the advanced cancer negative impact on long term outcome after any treatment (surgery, chemotherapy) is a limit for the clinical application of multidisciplinary treatments. Aim of this study is to review the possible different approaches to improve the clinical results, either as tumour response or overall survival, using an association of IL-2 with different chemotherapy procedures, in order to recover the locoregional and/or systemic immunodeficency. Several literature studies are worth of consideration not only for the biological activity reported, but also for the preliminary clinical results. At our Department, we have started a clinical experience in order to verify and confirm the results reported in these studies. The preliminary results seem to confirm an increase of chemotherapy activity obtained with an association of IL-2 immunotherapy with systemic therapy procedures and mainly with locoregional therapeutic programs. PMID- 15876992 TI - The ''central'' quadrantectomy. Introducing a new reconstruction technique after excision of the central sub- and periareolar portion of the breast. AB - A new method is presented for the closure of a central cylindrical breast exeresis, embracing, full thickness, the skin and the gland. For this reconstruction, performed without altering the original shape of the breast, without removing any additional healthy tissue, we used a skin-gland flap rotated from the inferior quadrant to correct the defect. PMID- 15876993 TI - [Laparoscopic surgery: from clinic to legal medicine]. AB - The laparoscopic technique introduced a new way of operating but inevitably causing new problems for the surgeon. After a comprehensive review of the history and the evolution of laparoscopic surgery from its beginning, the technical aspects of minimally invasive surgery and its fields of application are described. The close dependence on instruments and technology is emphasized. A detailed analysis of the advantages and limitations of laparoscopy is made with emphasis on the importance of a risk-benefit evaluation by the health care provider. Of key importance is to obtain a detailed and clear informed consent. The medico legal aspects of intraoperative complications and the liability of the surgical team in case of patients' injury or death are examined. However, it is always necessary to consider if the potential complications are predictable and/or preventable in accordance to the parameters of negligence, imprudence and lack of knowledge. The same criteria have to be applied to assure compliance with the preventive sanitary rules and that the conversion to laparotomy has been promptly carried out. PMID- 15876994 TI - Focus on current controversies in endovascular repair of abdominal aortic aneurysms. PMID- 15876995 TI - Thrombophilia and venous thromboembolism. International consensus statement. Guidelines according to scientific evidence. AB - Thrombophilia is the term now used to describe predisposition to increased risk of venous and occasionally arterial thromboembolism due to hematological abnormalities. It can be a multifactorial disorder where congenital defects of anticoagulant or procoagulant factors may be combined with acquired hematological abnormalities. It should be considered in patients with a documented unexplained thrombotic episode or a positive family history. The aim of this document is to provide guidelines for investigation and management of patients with thrombophilia in the presence or absence of venous thromboembolism (VTE). PMID- 15876996 TI - A critical appraisal of non-invasive diagnosis and exclusion of deep vein thrombosis and pulmonary embolism in outpatients with suspected deep vein thrombosis or pulmonary embolism: how many tests do we need? AB - The requirement for a safe diagnostic strategy should be based on an overall posttest incidence of venous thromboembolism of less than 1% during 3 month follow-up. Compression ultrasonography (CUS) has a negative predictive value (NPV) of 97% to 98% indicating the need of repeated CUS testing. Serial CUS testing is safe but you have to repeat 100 CUS to find 1 or 2 CUS positive for deep vein thrombosis (DVT), which is not cost-effective indicating the need to improve the diagnostic work-up of DVT by the use of clinical score assessment and D-dimer testing. The combination of a less sensitive D-dimer test (SimpliRed) and low clinical score does not, whereas the combination of a sensitive D-dimer test (ELISA VIDAS or Tinaquant) and low clinical score does safely exclude DVT without the need of CUS. The combination of a first negative CUS and a negative less sensitive D-dimer test (SimpliRed) or a sensitive ELISA D-dimer at a higher cut off level of 1,000 ng/ml safely excludes DVT with a NPV of > 99% without the need to repeated CUS in about 60%. The sequential use of a sensitive D-dimer and clinical score assessment will safely reduce the need for CUS testing by 40% to 60%. Large prospective outcome studies demonstrate that one negative examination with complete duplex color ultrasonography (CCUS) of the proximal and distal veins of the affected leg with suspected DVT is safe to withhold anticoagulant treatment with a NPV of 99.5%. This indicates that CCUS is equal or superior to serial CUS or the combined use of clinical score, D-dimer testing and CUS. Pulmonary angiography is the gold standard for segmental pulmonary embolism (PE) but not for subsegmental PE. A normal perfusion lung scan and a normal rapid ELISA VIDAS D-dimer test safely exclude PE. Helical spiral CT detects all clinically relevant PE and a large number of alternative diagnoses in symptomatic patients with a non-diagnostic ventilation perfusion scan (VP-scan) or a high probability VP-scan. Single-slice helical CT as the primary diagnostic test in patients with suspected PE in 5 retrospective studies and in 3 prospective management studies indicate that the NPV of a normal helical spiral CT, a negative CUS of the legs together with a low or intermediate pretest clinical probability is 99%. Helical spiral CT can replace both the VP-scan and pulmonary angiography to safely rule in and out PE. The combination of clinical assessment, a rapid ELISA VIDAS D-dimer followed by CUS will reduce the need for helical spiral CT by 40% to 50%. PMID- 15876997 TI - Pharmacodynamic effects of low molecular weight heparin in obese subjects following subcutaneous administration of 75 IU/kg on plasma tissue factor pathway inhibitor and nitric oxide. AB - AIM: Pharmacodynamic effects of the low molecular weight heparin tinzaparin on plasma levels of tissue factor pathway inhibitor (TFPI) and nitric oxide (NO) were compared in obese subjects, as well as in normal healthy controls. METHODS: Obese (n = 13) patients received a single 75 IU/kg SC injection (the deep vein thrombosis prophylaxis dose) of tinzaparin. Blood samples were obtained pre- and postadministration of drug and at different intervals over 24 h and assayed for total TFPI and NO stable metabolites (nitrates and nitrites) plasma levels, using a specific immunoassay and calorimetric methods. RESULTS: Mean maximum plasma TFPI levels approached 150-230 ng/ml at the 0.8 h and up to 5 h posttinzaparin dose compared to basal TFPI levels of 35-90 ng/mL. Plasma TFPI levels were still about 2-fold above basal levels at 12 h and fell to basal levels at 16 h after tinzaparin dose. Basal plasma levels of NO, but not TFPI, were significantly lower (P < 0.01) in obese patients compared to controls. Similar TFPI (3-fold above basal at peak) and NO pharmacodynamic profiles for tinzaparin at 75 anti-Xa IU/kg were demonstrated in obese and in normal healthy subjects. Plasma NO (nitrate + nitrite) showed a lag time of about 5-6 h posttinzaparin followed by a steady increase with a peak at 12-15 h and slow decline with a significant residual level at 24 h in obese and healthy subjects. CONCLUSIONS: Data suggest a normal responsiveness of vascular endothelial cells and other cellular compartments to tinzaparin with regard to the pharmacodynamic profiles of plasma TFPI and NO in obese subjects. PMID- 15876998 TI - Circulating markers of inflammation are related to carotid artery atherosclerosis. AB - AIM: The role of inflammation in atherothrombotic disorders is becoming increasingly recognized. The present study prospectively investigates relationships between inflammatory markers and hemostatic variables, and non invasive measures of carotid artery atherosclerosis. METHODS: Markers of hemostasis (sP-selectin and fibrinogen), cytokines (IL-6, IL-8, TNF-a and MCP-1), inflammatory variables (hsCRP, SAA and calprotectin) and cell adhesion molecules (ICAM-1 and VCAM-1) as well as ultrasonography of the carotid arteries were assessed in 111 consecutive outpatients with manifest or suspected coronary artery disease (CAD). RESULTS: Thirty-eight patients with manifest cardiovascular disease had higher IL-6 (P < 0.01) but not hsCRP levels. Higher levels of IL-6, calprotectin and VCAM-1 (all P < 0.05) were found in 35 patients with carotid plaques. In the whole study population (n = 109) an increased common carotid artery lumen diameter (LD) and cross sectional intima-media area (CIMA) was related to higher IL-6, IL-8 and MCP-1 levels (all P < 0.05), and increased LD also to higher hsCRP, calprotectin (both P < 0.05), sP-selectin and fibrinogen levels (both P < 0.01). Both LD and CIMA were related to VCAM-1 (both P < 0.01), but not to ICAM-1 levels. The intima-media thickness of the carotid artery was only positively related to MCP-1 levels (P < 0.05). Only the relation between IL 6 and LD remained significant after adjustment for age, gender, body mass index, smoking status or present lipid-lowering treatment. CONCLUSIONS: Several biomarkers of inflammation are related to ultrasonographic measures of carotid artery atherosclerosis in patients with moderate to high prevalence of CAD. IL-6 seems to be an independent and useful biomarker of atherosclerosis in this group of patients. PMID- 15876999 TI - Relationship between endothelial dysfunction, intima media thickness and cardiovascular risk factors in asymptomatic subjects. AB - AIM: The aim of the study was to evaluate endothelial function and intima media thickness (IMT) in relation to cardiovascular risk factors (RF). METHODS: We enrolled 113 patients, mean age 62 +/- 12 years; patients underwent: anamnesis, physical examination, measurement of body weight and height and blood pressure. Biochemistry variables were also measured: total cholesterol, high- and low density lipoprotein cholesterol (HDL-C and LDL-C), triglycerides and glycemia. Vascular echography was performed to analyze flow mediated vasodilatation (FMD) at the brachial artery and IMT of the carotid and femoral arteries. RESULTS: Compared with patients without RF, patients with cardiovascular RF showed an impaired FMD (p < 0.05) and higher values of mean carotid IMT (p = 0.03). Age (p < 0.005) and diabetes (p < 0.05) were directly correlated with carotid IMT, while femoral IMT is correlated with age (p < 0.005) and male gender (p < 0.02). Regarding the relationship between endothelial function cardiovascular RF, we showed an inverse linear correlation between systolic blood pressure (p < 0.005), smoking (p < 0.05) and FMD, and concerning biochemical parameters, we founded that total cholesterol (p < 0.05) and LDL-C plasma levels (p < 0.005) were inversely correlated with FMD. Finally, we showed a lower FMD in patients with carotid and femoral IMT in comparison with patients without peripheral atherosclerosis (p = 0.01). CONCLUSIONS: The present data indicate that cardiovascular RF are associated with impaired endothelial function and increased IMT, and that the presence of carotid and femoral IMT is significantly correlated with endothelial dysfunction. PMID- 15877000 TI - Suboptimal treatment of risk factor for atherosclerosis in critical limb ischemia. AB - AIM: The epidemiology of critical limb ischemia (CLI) is insufficiently studied, and treatment of risk factors for atherosclerosis has received less attention in CLI patients than in patients with coronary or precerebral atherosclerosis. The aim of this study was to establish the incidence of CLI and the quality of risk factor treatment in Swedish CLI patients. METHODS: During 14 months, 316 consecutive CLI patients were referred to the Malmo Department of Vascular Diseases. Two hundred and fifty-nine (82%) consented to evaluation of intercurrent disease, medication, ankle and arm blood pressures (BP), plasma glucose and lipid levels, p-homocysteine, cardiolipin antibodies and activated protein C (APC)-resistance. RESULTS: The incidence of CLI was 38/100,000 inhabitants/year. Patient age was 75 +/- 10 years, and BP 147 +/- 26/75 +/- 14 mmHg. Systolic or diastolic BP above recommended levels (140/90 mmHg) occurred in 137 (53%) patients. P-cholesterol was 4.8+/-1.2 mMol/L, but cholesterol above recommended level (5 mMol/L) or LDL above recommended level (3 mMol/L) occurred in 125 (48%) patients. Only 24% of patients met national recommendations for both BP and lipid levels. Diabetes mellitus was previously known in 123 (47%) patients, and another 12 (5%) patients showed diabetic fasting glucose levels during the hospital stay. Eighty-four (32%) patients were active, and 72 (28%) were former smokers. Myocardial infarction or angina pectoris had previously been diagnosed in 123 (47%) patients. P-homocysteine was 17 +/- 7 _mol/l, cardiolipin antibodies occurred in 71 (27%) and APC-resistance in 34 (13%) patients. CONCLUSIONS: Patients with CLI show high comorbidity in vascular diseases and high prevalence of modifiable risk factors for atherosclerotic vascular disease. The use of evidence-based medical therapy is suboptimal in this high-risk group. PMID- 15877001 TI - Short-term and long-term effects of one-week treatment with intravenous iloprost in critical limb ischemia patients (Leriche-Fontaine stage III and IV). AB - AIM: Iloprost, usually administered through intravenous infusion for 6 hours per day for at least 21 days, is the main medical treatment for critical limb ischemia in patients unsuitable for surgical or endovascular approach. We evaluated the tolerance and the short-term and long-term effects of a single 1 week treatment in critical limb ischemia patients. METHODS: Twenty-nine patients in Leriche-Fontaine III and IV stage were treated with iloprost infusions for 16 hours per day for 7 days, achieving a maximal dose of 1.5 ng/kg/min. Tolerance and clinical assessment after treatment discontinuation and after 1 and 6 months were recorded; clinical evaluation (rest pain, trophic lesions), ankle/brachial pressure index (ABPI) and treadmill exercise test were performed before, immediately after treatment and after 1 and 6 months. RESULTS: No discontinuation of treatment occurred because of intolerance to iloprost. At the end of the treatment 69% of patients were responders, 55.2% at 1 month, 37.9% after 6 months. ABPI and treadmill maximum walking distance were improved by the treatment at every timepoint. After 6 months 10.3% mortality and 3.4% major amputation rates were recorded. There was a higher percentage of non-responders amongst women vs men, in diabetic patients vs non diabetic and in stage IV patients vs stage III. CONCLUSIONS: One-week treatment with iloprost is safe and effective in both Leriche-Fontaine stage III and IV patients. Clinical effects are persistent over time, often lasting up to the 6th month, similarly to the commonly used 28-day treatment, with clear implications in terms of patient's compliance and medical cost containment. PMID- 15877002 TI - What is the best management for abdominal aortic aneurysm in patients at high surgical risk? A single-center review. AB - AIM: To determine the best treatment for high-risk patients with abdominal aortic aneurysms (AAA). METHODS: We reviewed a prospective database of all patients who underwent conventional (OPEN) or endovascular aneurysm repair (EVAR) between January 1998 and December 2002. Patients were preoperatively classified according to the American Society of Anesthesiology (ASA). Comorbidities and medical risk factors were categorized according to the Ad Hoc Committee on Reporting Standards. Perioperative mortality and morbidity rates were analyzed according to the type of surgical procedure (OPEN vs EVAR) and ASA class. Patients in ASA classes I and II were excluded. Continuous data were expressed as mean +/- standard deviation. All data were calculated using the cumulated actuarial method of event outcome probability. Kaplan-Meier curves were constructed and the log rank statistic and chi squared test were used for comparative data. P values less than 0.05 were considered to indicate statistical significance. RESULTS: Of the total 375 patients who underwent AAA repair, 168 (45%) belonged in ASA classes III and IV (85 submitted OPEN and 83 EVAR to repair). Among general risk factors only coronary artery disease differed significantly between the 4 groups (P = 0.04). The Bonferroni correction identified a statistically significant difference between ASA classes III and IV for the OPEN technique and for EVAR (P = 0.007 and P = 0.012). Neither 30-day morbidity or mortality differed significantly according to ASA class and surgical technique. The median follow-up was 19 months (range 5-60 months). The overall survival was 78% at 60 months. Survival rates during follow-up differed significantly in the 2 risk classes (ASA III 5/123, 4% vs ASA IV 9/38, 24%), (P = 0.0001). The deaths in the ASA class 4 patients (12/14; 86%) were caused by preexisting medical comorbidities (in 9 patients cardiovascular, in 1 cancer and in 2 cirrhosis). CONCLUSIONS: Except patients with small aneurysms (< 6 cm), in whom the risk of death at 1-year due to comorbidities exceeds the risk of a ruptured aneurysm, all patients at high surgical risk (ASA class IV) benefit from AAA repair. Patients with small aneurysms must undergo strict surveillance to assess growth and aneurysmal wall changes to prevent unexpected rupture. PMID- 15877003 TI - Transilluminated powered phlebectomy. Mid-term clinical experience. AB - AIM: Transilluminated powered phlebectomy (TIPP) was first described in 1996 by Sptiz et al. and was designed to allow minimally invasive surgical treatment of varicose veins (VV). We report our updated experience with TIPP technique. METHODS: Between January 2001 and February 2004, 84 patients underwent treatment by TIPP technique for primary symptomatic VV. Saphenofemoral junction with complete stripping of the great saphenous vein was performed in all patients. Incompetent perforating veins was ligated and prominent VV were ablated with TIPP technique. RESULTS: Mean age of patients was 50.6 years (range 29-79 years) and most of patients were women (73%). The major varicose vein risk factors were standing position and parity. Heaviness (62%), pain (57%) and unsightly veins (30%) were the most common indications for surgery. The mean number of surgical incisions was 6 (range 3-10), the average operative time was 59 min (range 30-100 min) and the mean cosmetic score (out of 10) at 6 weeks was 8 (range 2-10). The mean pain score (out of 10) was at 2 days, 7 days and 6 weeks was 5, 3 and 0, respectively. All the complications were documented. CONCLUSIONS: The TIPP technique is safe without any adverse events, presents advantages and inconvenient which are discussed in this paper. PMID- 15877004 TI - Transcutaneous oxygen pressure measurements (tcpO2) at ankle during exercise in arterial claudication. AB - AIM: Although a time consuming technique, tcpO2 provides complementary information as compared to other tests. Simultaneous recording of systemic and peripheral oxygen pressure changes with exercise could be interesting to confirm that local hypoxemia is of arterial origin, but accuracy versus gold standard arteriography and objectively determined cut-off points to be used in arterial claudication at the ankle are not reported. METHODS: EXPERIMENTAL DESIGN: retrospective plus prospective study. SETTING: institutional practice, ambulatory care. PATIENTS: 100 patients suffering stage 2 claudication (group A) were retrospectively studied to objectively define cut-off points derived from tcpO2 recordings to be used in exercise testing. Then, applicability and reproducibility of these cut-off points were analysed prospectively in another 50 patients (group B). INTERVENTION: tcpO2 was measured on both calves and with a chest reference electrode. Arteriography on each side was quoted positive for a diameter stenosis superior to 75% or occlusion on the aorto-popliteal axis or of all-3-calf arteries. RESULTS: The best performance was obtained with tcpO2 changes from rest at the calf normalised to eventual chest changes (DROP) during or following the treadmill test. Optimal cut-off point determined through ROC curve analysis for DROP was -15 mmHg in group A. Applying this cut-off point in group B provided a 86/84% sensitivity/specificity and showed excellent reproducibility. CONCLUSIONS: TcpO2 measurement on the calf during exercise could be useful in a selected population of patients with claudication of questionable vascular origin and/or when other non-invasive investigations cannot be performed. PMID- 15877005 TI - Arterial distensibility, intima media thickness and pulse wave velocity after renal transplantation and in dialysis normotensive patients. AB - AIM: Structural and mechanical properties of the arterial wall are altered in patients with renal failure. The ageing process of the arterial wall appears to be accelerated in patients with end-stage renal failure. The mechanisms responsible for reduced arterial compliance and distensibility in dialysis patients and renal transplant recipients without hypertension remain to be evaluated. METHODS: Thirty-five normotensive dialysis patients (D), 35 normotensive renal transplant recipients (T) and 35 healthy volunteers (N) matched for age, sex and blood pressure as controls were enrolled into the study. The arterial blood pressure of all patients was < 140/90 mmHg. The dialysis patients and renal transplant recipients were eligible for the study if the serum creatinine level was < 2 mg/dL. In all subjects, fasting concentrations of serum creatinine, total cholesterol, HDL-cholesterol, LDL-cholesterol and hemoglobin and glucose were determined at enrollment to the study. Blood pressure was measured using an automatic sphygmomanometer. Pulse wave velocity (PWV) was evaluated using non invasive automatic Complior device. The vessel wall properties of the left common carotid artery were studied using multigate pulsed Doppler system. With this method, the end-diastolic diameter (d) and the systolic increase of vessel diameter (distension DELTAd) were measured. From these data the relative systolic increase of vessel diameter (DELTAd/d) and the arterial wall distensibility coefficient (DC) were calculated. RESULTS: Systolic blood pressure (SBP) and central pulse pressure (CPP) were significantly higher in T than in D and N group, respectively 138 +/- 18 mmHg and 59 +/- 16 mmHg vs 128 +/- 13 mmHg and 49 +/- 12 mmHg and 132.12 mmHg and 51 +/- 10 mmHg. The d did not change significantly between all groups. The distension DELTAd was significantly lower in patients group D and T, respectively 466 +/- 38 microm and 511 +/- 37 microm than in controls. Similarly DELTAd/d was in these groups significantly lower than in healthy volunteers, respectively D 6.33 +/- 0.5%, T 6.9 +/- 0.4% vs N 9.15 +/- 0.5%. DC was also significantly lower in D and T than in N groups, respectively D 17.91 +/- 1.5 10-3/kPa and T 18.92 +/- 1.3 10-3/kPa and N 24.28 +/ 0.51-3/kPa. Significant differences were found in the increase of the intima media thickness (IMT) of carotid artery for dialyzed patients and renal transplant recipients in contrast to the control group, but there were no differences between the patients. PWV in both patient groups was statistically significant higher than in control group correspondingly D 11.1 +/- 1.03 m/s and T 13.3 +/- 1.13 m/s, N 9.4 +/- 0.89 m/s. There was a significant correlation between the change of DC, PWV and CPP in T group (n = 35; r = -0.43; P < 0.01 and n = 35; r = 0.48; P < 0.05). In the T group also an important correlation between PWV and IMT complex (n = 35; r = 0.49, P < 0.001) was found. CONCLUSIONS: The elastic and structural properties of arterial wall in dialysis patients and renal recipients are decreased. End-stage renal disease accelerates arterial stiffening despite of arteriosclerosis and hypertension. Renal transplantation does not reverse loss of elastic and morphologic properties of arteries found in patients with end-stage renal insufficiency. PMID- 15877006 TI - Endoluminal repair of anastomotic false popliteal aneurysm using the Wallgraft endoprosthesis. AB - The aim of this study was to present the endoluminal repair of a false anastomotic popliteal aneurysm on a previous polytetrafluoroethylene (PTFE) above knee bypass using a Wallgraft endoprosthesis. A 53-year-old man who underwent a left femoro-popliteal above knee PTFE bypass 13 years before was admitted with a painful pulsatile mass in the above knee area. Nine months previously he developed sudden pain around the knee extending to the foot associated with coldness and numbness after he was handling some fishing gear in a crouching position for about 1 hour, but this episode spontaneously resolved. Duplex scanning and angiography revealed a 3 x 2.5 cm false aneurysm, which was successfully treated by deploying a Wallgraft endoprosthesis as the patient declined surgical repair. The graft was detected as being occluded on the 3-month follow-up but no further action was taken because the patient experienced only non-limiting claudication and he refused again surgical treatment. Endoluminal repair of perianastomotic false aneurysms in the popliteal artery with Wallgraft endoprosthesis seems feasible and safe, but until its durability is validated in larger series surgical repair remains the treatment of choice. PMID- 15877007 TI - Ruptured abdominal aortic aneurysm secondary to tuberculous spondylitis. AB - A 69-year-old man was admitted with low back pain and signs of nerve root compression. A computed tomography (CT) scan showed abscess formation in the left psoas region, spondylodiscitis L3-L4 and a ruptured abdominal aortic aneurysm. The aortic aneurysm was replaced with a bifurcated vascular graft. One week later, laminectomy at the L4-level was done. In a small abscess, Mycobacterium bovis was found. The condition was considered to be a mycobacterial spondylitis secondary to BCG instillations of the urinary bladder for carcinoma. The patient received antituberculous medication for 9 months. Subsequently bone transplantation and internal fixation of the spine became necessary. Three years after surgery he is in good condition and there are no signs of graft infection on CT. Spondylitis and mycotic aortic aneurysm should be kept in mind in patients who have been treated for carcinoma of the bladder with BCG instillations. PMID- 15877008 TI - Ex vivo repair of renal artery aneurysm associated with repairing of abdominal aortic aneurysm. Case report. AB - A case of right renal artery distal aneurysm associated with juxtarenal abdominal aortic aneurysm in a 75-year-old male, who presented with abdominal and back pain and chronic renal failure, is reported. The abdominal aortic aneurysm was repaired with a bifurcated Dacron graft. The right kidney was simultaneously explanted, ex vivo reconstruction of the renal artery with PTFE graft was performed, followed by autotransplantation of the kidney into the right iliac fossa. In the postoperative course the renal function returned to normal. PMID- 15877009 TI - Growth hormone and the cardiovascular function. AB - In this review, the great importance of growth hormone (GH) for the maintenance of cardiac function in adult life is discussed. Physiological effects of GH are discussed as well as the cardiac dysfunction caused both by GH excess (acromegaly) and by GH deficiency in adult hypopituitary patients. In both acromegaly and adult GH deficiency, there is also increased cardiovascular morbidity and mortality. Finally, the effect of GH treatment in heart failure is discussed. PMID- 15877010 TI - Androgens and the skeleton. AB - Loss of estrogens or androgens causes bone loss by increasing the rate of bone remodeling, and also causes an imbalance between resorption and formation by prolonging the lifespan of osteoclasts and shortening the lifespan of osteoblasts. Conversely, treatment with androgens, as well as estrogens, maintains cancellous bone mass and integrity, regardless of age or sex. Both androgens, via the androgen receptor (AR), and estrogens, via the estrogen receptors (ERs) can exert these effects, but the relative contribution of these 2 pathways remains uncertain. Androgens, like estrogens, stimulate endochondral bone formation at the start of puberty, whereas they induce epiphyseal closure at the end of puberty, thus, they have a biphasic effect. Androgen action on the growth plate is, however, clearly mediated via aromatization into estrogens and interaction with ER alpha. Androgens increase, while estrogens decrease radial growth. This differential effect of the sex steroids may be important because bone strength in males seems to be determined by higher periosteal bone formation and, therefore, greater bone dimensions. Experiments in mice suggest that both the AR and ER alpha pathways are involved in androgen action on radial bone growth. ER beta may mediate growth-limiting effects of estrogens in the female but does not seem to be involved in the regulation of bone size in males. In conclusion, androgens may protect men against osteoporosis via maintenance of cancellous bone mass and expansion of cortical bone. This androgen action on bone is mediated by the AR and ER alpha. PMID- 15877011 TI - Hormone replacement therapy in postmenopausal women. AB - From the endocrine point of view, menopause is considered a deficiency state and menopausal hormone replacement therapy (HRT) regarded as restoring the premenopausal endocrine milieu. Millions of healthy postmenopausal women were taking HRT in late 1990's many in the absence of menopausal symptoms. The major benefit from HRT was considered to be cardiovascular protection and also protection against osteoporosis and Alzheimer's Disease. The Women's Health Initiative (WHI) trial and other studies published since 2002 fundamentally changed our understanding of risks and benefits associated with HRT. This review discusses the effects of HRT on menopausal symptoms, cognitive function, cardiovascular disease, osteoporosis and also breast and bowel cancer. PMID- 15877012 TI - 11Beta-hydroxysteroid dehydrogenase type 1 in human disease: a novel therapeutic target. AB - Patients with cortisol excess, Cushing's syndrome, develop a classical phenotype characterized by central obesity, hypertension, and increased cardiovascular mortality. Whilst this observation points to the importance of glucocorticoids, circulating cortisol excess is rare and does not explain the pathogenesis of many common conditions. At a tissue specific level, the enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) locally regenerates active cortisol from inactive cortisone amplifying glucocorticoid receptor activation in the context of normal circulating cortisol levels. Increased 11beta-HSD1 activity and expression have been implicated in the pathogenesis of many common conditions including, obesity, insulin resistance, the metabolic syndrome, polycystic ovarian syndrome, osteoporosis and glaucoma. Furthermore, selective 11beta-HSD1 inhibition has been proposed as a novel therapeutic strategy in many of these conditions. Here we review the role of 11beta-HSD1 in human disease and discuss the impact of selective 11beta-HSD1 inhibition. PMID- 15877013 TI - Disinfection of endoscopes from Helicobacter pylori-positive subjects: evaluation of the effectiveness of the Chinese Calijing disinfection kit. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness of the Calijing disinfection kit (an endoscope disinfection method used in Chinese hospitals) in eradicating Helicobacter pylori and assess whether use of the kit in 1994 during endoscopies in the Shandong Intervention Trial (SIT), Shandong, China, could have resulted in iatrogenic transmission of H pylori . METHODS: Bacterial culture studies at the Veterans Affairs Medical Center, Houston, Texas, using endoscopes and forceps from 49 H pylori -positive patients were performed on contaminated endoscopes before and after disinfection with the Calijing kit. RESULTS: At least 1 endoscope culture site was H pylori positive in 39 of 49 (79.6%) specimens predisinfection, whereas H pylori was not isolated from any endoscopic culture site postdisinfection. Non- H pylori bacteria and fungi were recovered from 22.6% of the postdisinfection cultures. CONCLUSION: Although no viable H pylori were recovered following the disinfection procedures, levels of H pylori below the detection threshold of the bacteriologic assay may have contributed to an increase in H pylori seroprevalence noted in the SIT. In addition, the kit was unable to provide disinfection against non- H pylori organisms, suggesting the need to adhere to internationally accepted disinfection procedures for endoscope reprocessing. PMID- 15877014 TI - Real-time monitoring in managing the decontamination of flexible gastrointestinal endoscopes. AB - OBJECTIVES: To evaluate the efficacy of endoscope reprocessing using bacteriologic methods and adenosine triphosphate (ATP) bioluminescence and to evaluate the potential benefits of the latter in managing the process. SETTING: An endoscopy unit in each of 2 acute district general hospitals in the United Kingdom. METHODS: Following visual observation, 8 locations important in the decontamination of gastrointestinal (GI) endoscopes were monitored during reprocessing by bacteriologic sampling (paddle method) and ATP bioluminescence. RESULTS: Endoscope reprocessing was conducted in accordance with British Society of Gastroenterology guidelines. Bacteriologic tests required 24 hours for results to be obtained; ATP tests were completed within 2 minutes. Of the 504 sites tested throughout the decontamination process, bacterial growth was recorded on 32 occasions, 3 from the end product. Throughout the decontamination process, ATP tests obtained 95 results above benchmark values and 1 from the end product. CONCLUSIONS: The bacteriologic results after terminal disinfection indicated that the reprocessing was carried out effectively overall. ATP results prior to disinfection indicated that manual cleaning might not have always been performed adequately and that this could have impaired the disinfection process. ATP provided a rapid means of assessing the efficacy of cleaning steps prior to terminal disinfection. PMID- 15877015 TI - A unique water optional health care personnel handwash provides antimicrobial persistence and residual effects while decreasing the need for additional products. AB - BACKGROUND: The Centers for Disease Control and Prevention (CDC) has published guidelines for hand hygiene practices, recommending a handwash regimen that alternates between waterless alcohol products and antimicrobial or nonantimicrobial soap and water. The advent of an alcohol-based product that can be used with or without water (ie, water optional) to decontaminate the hands while providing immediacy of kill and antimicrobial persistence could reduce the confusion associated with handwash guidelines. Such a product has been developed, is alcohol-based (61%), and zinc pyrithione (ZPT) preserved (61% alcohol-ZPT) and has proven to be fully compliant with the Food and Drug Administration (FDA) and CDC guidelines. METHODS: FDA-required testing of the 61% alcohol-ZPT product for the health care personnel handwash indication was performed as outlined in the Tentative Final Monograph (TFM) for Health-Care Antiseptic Drug Products, employing waterless and water-aided product applications. It was next assessed for antimicrobial persistence and residual effects by comparing it, in separate waterless and water-aided applications, with commonly available handwashes containing various antimicrobials in a 5-day study employing 49 subjects, in which samples were collected immediately and at 4 hours and 8 hours postapplication. The skin conditioning properties of this formulation were investigated via appropriate methods. RESULTS: The 61% alcohol-ZPT product easily produced >3.0 log 10 reduction in the indicator strain ( Serratia marcescens ) following the first wash, exceeding the 2.0 log 10 FDA requirement. This level of performance was maintained through the tenth wash, surpassing the 3.0 log 10 FDA requirement for the handwash indication. For the assessment of persistence and residual effect in the waterless mode, the water-optional, 61% alcohol-ZPT product consistently produced log 10 reductions of nearly 3.5 or greater at every point over the entire study period. In the water-aided configuration, similar results were obtained as log 10 reductions of 2.5 were observed. The formulation is nonirritating, actually contributing to hand skin condition. CONCLUSIONS: The 61% alcohol-ZPT product exceeds all FDA criteria for the health care personnel handwash indication and is a significant advancement in the concept of skin antisepsis. It represents a single product suitable for use in all hand hygiene settings, demonstrating improved antimicrobial persistence and residual effects. The 61% alcohol-ZPT formulation contributes positively to overall hand conditioning, and a previously reported study has documented it to be virucidal for several DNA and RNA viruses. PMID- 15877016 TI - Guidance on public reporting of healthcare-associated infections: recommendations of the Healthcare Infection Control Practices Advisory Committee. AB - Since 2002, 4 states have enacted legislation that requires health care organizations to publicly disclose health care-associated infection (HAI) rates. Similar legislative efforts are underway in several other states. Advocates of mandatory public reporting of HAIs believe that making such information publicly available will enable consumers to make more informed choices about their health care and improve overall health care quality by reducing HAIs. Further, they believe that patients have a right to know this information. However, others have expressed concern that the reliability of public reporting systems may be compromised by institutional variability in the definitions used for HAIs, or in the methods and resources used to identify HAIs. Presently, there is insufficient evidence on the merits and limitations of an HAI public reporting system. Therefore, the Healthcare Infection Control Practices Advisory Committee (HICPAC) has not recommended for or against mandatory public reporting of HAI rates. However, HICPAC has developed this guidance document based on established principles for public health and HAI reporting systems. This document is intended to assist policymakers, program planners, consumer advocacy organizations, and others tasked with designing and implementing public reporting systems for HAIs. The document provides a framework for legislators, but does not provide model legislation. HICPAC recommends that persons who design and implement such systems 1) use established public health surveillance methods when designing and implementing mandatory HAI reporting systems; 2) create multidisciplinary advisory panels, including persons with expertise in the prevention and control of HAIs, to monitor the planning and oversight of HAI public reporting systems; 3) choose appropriate process and outcome measures based on facility type and phase in measures to allow time for facilities to adapt and to permit ongoing evaluation of data validity; and 4) provide regular and confidential feedback of performance data to healthcare providers. Specifically, HICPAC recommends that states establishing public reporting systems for HAIs select one or more of the following process or outcome measures as appropriate for hospitals or long-term care facilities in their jurisdictions: 1) central-line insertion practices; 2) surgical antimicrobial prophylaxis; 3) influenza vaccination coverage among patients and healthcare personnel; 4) central line-associated bloodstream infections; and 5) surgical site infections following selected operations. HICPAC will update these recommendations as more research and experience become available. PMID- 15877017 TI - A brief report on the normal range of forehead temperature as determined by noncontact, handheld, infrared thermometer. AB - BACKGROUND: Noncontact forehead temperature measurement by handheld infrared thermometer was used as a screening tool for fever. However, the accuracy data and normal range of forehead temperature determined by this method were not available. METHODS: The temperature readings from 3 handheld infrared thermometers were validated against an electronic thermometer. Normal range of forehead temperature was determined by measuring the forehead temperature in 1000 apparently healthy subjects. RESULTS: Significant differences were detected in readings obtained by the 3 different handheld infrared thermometers (analysis of covariance, P < .001) The most accurate one was chosen, and the normal range of forehead temperature in 1000 subjects detected by this method was 31.0 degrees C to 35.6 degrees C. CONCLUSIONS: Our study shows that commercially available, handheld infrared thermometers require individual validation. Forehead temperature in excess of 35.6 degrees C is suggestive of fever. Further studies are required to confirm accuracy of this value in detecting fever. PMID- 15877018 TI - Colonization of personal digital assistants used in a health care setting. AB - BACKGROUND: The use of personal digital assistants (PDAs) by health care workers is increasing. Increasing rates of infection in our institution led to the question of whether PDAs were colonized with pathogenic organisms. METHODS: Specimens for culture were obtained from PDAs used at our institution, and surveys were distributed to the users to determine factors predisposing to colonization. RESULTS: Forty percent of PDAs had growth on culture. The most common organism detected on 27 of 82 PDAs was coagulase-negative Staphylococcus (82%). No isolates of methicillin-resistant Staphylococcus aureus or vancomycin resistant enterococci were detected. Colonization was more common on PDAs that had undergone previous cleaning. No other predisposing factors to colonization were found. CONCLUSIONS: PDAs are frequently colonized with typical skin organisms and less commonly with pathogenic organisms. Whether PDAs used in the health care setting serve as vectors for nosocomial infection is not determined. PMID- 15877019 TI - Computer equipment used in patient care within a multihospital system: recommendations for cleaning and disinfection. AB - Computer hardware has been implicated as a potential reservoir for infectious agents. Leaders of a 22-hospital system, which spans North America and serves pediatric patients with orthopedic or severe burns, sought to develop recommendations for the cleaning and disinfection of computer hardware within its myriad patient care venues. A task force comprising representatives from infection control, medical affairs, information services, and outcomes management departments was formed. Following a review of the literature and of procedures within the 22 hospitals, criteria for cleaning and disinfection were established and recommendations made. The recommendations are consistent with general environmental infection control cleaning and disinfection guidelines, yet flexible enough to be applicable to the different locales, different computer and cleaning products available, and different patient populations served within this large hospital system. PMID- 15877020 TI - A 5-year experience with influenza prevention and containment in a program of all inclusive care for elderly adults. AB - BACKGROUND: Influenza causes mortality and morbidity in the frail elderly population. Influenza prevention and mitigation models need to be developed for this population. METHODS: An observational study at a Program for All-Inclusive Care for the Elderly (PACE) during years 1999-2004. Participants were frail elderly adults who meet the state Medicaid requirements for nursing home placement. RESULTS: Over 91% of participants have been immunized yearly since the beginning of the program. Employee immunization has increased yearly from 61% to 90%, and caregivers known to be immunized increased from 9% to 62% over the last 4-year period. During 2 influenza seasons, we placed all our participants on ramantidine prophylaxis within a 2-week period. CONCLUSIONS: The incentives and flexibility offered by PACE have allowed our participants to enjoy much higher influenza immunization rates than experienced by other elderly adults while also prophylaxing employees and caregivers. Our systems have allowed us to prophylax all our participants within a short time during 2 influenza seasons. Nonetheless, many questions remain regarding the optimal way to decrease the burden of influenza in frail elderly adults. With adequate integration and supplementary financing, PACE programs throughout the United States could serve as laboratories to test candidate interventions. PMID- 15877021 TI - Understanding influenza vaccination attitudes at a Canadian cancer center. AB - BACKGROUND: Although influenza vaccination benefits both health care workers and their patients, participation by staff in vaccination programs is disappointingly low. Understanding health care worker perceptions and needs is essential for improving rates of vaccination. METHODS: A self-administered questionnaire was distributed to all staff at a Canadian cancer center. Information was sought on previous frequency of participation in influenza vaccination, as well as motivations, perceptions, and preferences. RESULTS: Three hundred sixty-three (70%) of 515 cancer center staff members responded. Twenty-two percent of staff were vaccinated 4 or 5 times in the past 5 years and were primarily motivated by the desire to protect their own health (81%). Forty-nine percent participated 1 to 3 times in the past 5 years, and this group had diverse knowledge and vaccine access needs. Twenty-nine percent received no vaccination in the previous 5 years because they believed the vaccine lacked efficacy (45%) or was harmful to health (19%). Moving from high to low levels of participation with influenza vaccination, the following trends were observed: increasing belief that vaccines cause illnesses or weaken the immune system, increasing belief that adverse effects of vaccination are underreported, and decreasing belief that vaccination programs are beneficial. CONCLUSIONS: Cancer center staff perceptions about influenza vaccination differ according to the past frequency of vaccine uptake. Strategies for promoting vaccination should be guided by these differences. PMID- 15877023 TI - Soft tissue lasers. PMID- 15877024 TI - Invisalign clinical trials needed. PMID- 15877025 TI - Buccal smile corridors. PMID- 15877027 TI - Functional matrix theory. PMID- 15877028 TI - Author's response to "Effect of cervical headgear". PMID- 15877030 TI - Class I occlusion a false goal? PMID- 15877032 TI - Obsequies for an opinion masquerading as fact. PMID- 15877033 TI - Rapid canine retraction and orthodontic treatment with dentoalveolar distraction osteogenesis. AB - PURPOSE: Duration of treatment is one of the things orthodontic patients complain about most. To shorten treatment time, a new technique of rapid canine retraction through distraction osteogenesis was introduced. The effects of dentoalveolar distraction on the dentofacial structures are presented in this article. MATERIAL: The study sample consisted of 20 maxillary canines in 10 growing or adult subjects (mean age, 16.53 years; range, 13.08-25.67 years). First premolars were extracted, the dentoalveolar distraction surgical procedure performed, and a custom-made intraoral, rigid, tooth-borne distraction device was placed. The canines were moved rapidly into the extraction sites in 8 to 14 days, at a rate of 0.8 mm per day. RESULTS: Full retraction of the canines was achieved in a mean time of 10.05 (+/-2.01) days. The anchorage teeth were able to withstand the retraction forces with minimal anchorage loss. The mean change in canine inclination was 13.15 degrees +/- 4.65 degrees, anterior face height and mandibular plane angle increased, and overjet decreased significantly at the end of dentoalveolar distraction. No clinical and radiographic evidence of complications, such as root fracture, root resorption, ankylosis, periodontal problems, and soft tissue dehiscence, was observed. Patients had minimal to moderate discomfort after the surgery. CONCLUSIONS: The dentoalveolar distraction technique is an innovative method that reduces overall orthodontic treatment time by nearly 50%, with no unfavorable effects on surrounding structures. PMID- 15877034 TI - Numerical simulation of canine retraction by sliding mechanics. AB - BACKGROUND: Bone remodeling laws have been used to simulate the movement of a single tooth, but the calculations for simulating the movement of several teeth simultaneously are time-consuming. The purpose of this article is to discuss a method that allows the simulation of more complex tooth movements. METHODS: A 3 dimensional finite element method was used to simulate the orthodontic tooth movement (retraction) of a maxillary canine by sliding mechanics and any associated movement of the anchor teeth. Absorption and apposition of the alveolar bone were produced in proportion to the stress of the periodontal ligament. RESULTS: In a reference case, the canine was retracted by a 2N force with 0.016-in square wire. The frictional coefficient between wire and bracket was 0.2. The movement of both the canine and the anchor teeth could be calculated with the elastic deformation of wire. The canine tipped during the initial unsteady state and then moved bodily during the steady state. It became upright when the orthodontic force was removed. The anchor teeth moved in the steady state and tipped in the mesial direction. The decrease in applied force by friction was about 70%. The tipping of the canine decreased when the wire size was increased or when the applied force was decreased. CONCLUSIONS: Simple assumptions were used in this calculation to simulate orthodontic tooth movements. The calculated results were reasonable in mechanical considerations. This method might enable one to estimate various tooth movements clinically. However, precise comparisons between calculated and clinical results, and the improvement of the calculation model, are left for a future study. PMID- 15877035 TI - Factors of importance for the development of dehiscences during labial movement of mandibular incisors: a retrospective study of adult orthodontic patients. AB - PURPOSE: Labial movement of mandibular incisors has traditionally been considered a risk factor for gingival recession. The aims of this study were to assess changes in prevalence and severity of gingival recession of mandibular incisors during orthodontic treatment of adults in whom the incisors had been moved labially and to identify parameters that could predict recession. MATERIAL: The sample consisted of 150 adult patients (aged 33.7 +/- 9.5 years, mean +/- SD) treated nonextraction with fixed appliances. Pretreatment overjet, overbite, degree of crowding, presence of tooth rotation, canine relationship, vertical face height, and position of the mandibular incisor to A-pogonion and mandibular lines were registered on study casts and lateral headfilms. Pretreatment gingival recession, width of keratinized gingiva, gingival biotype, gingival inflammation, and visible plaque accumulation were recorded, as was posttreatment gingival recession. Labial movement was determined by measuring pretreatment and posttreatment casts. Descriptive statistics were used to describe gingival recession at baseline and follow-up. Variables to be included in a logistic regression analysis as possible predictors of recession were identified with a bivariate correlation analysis. RESULTS: No significant increase in the mean gingival recession was observed during treatment. The prevalence of gingival recession greater than 0.1 mm increased from 21% before treatment to 35% after (P < .05). Only 2.8% of the subjects developed recession greater than 2 mm, and 5% of the pre-existing gingival recessions improved. The presence of baseline recession (P < .001), gingival biotype (P < .0179), and gingival inflammation (P < .003) were identified as possible predictors of recession. None of the orthodontic variables was significantly associated with recession. CONCLUSIONS: Gingival recession of mandibular incisors did not significantly increase during orthodontic treatment. After treatment, fewer than 10% of subjects had gingival recession greater than 2 mm, and, at follow-up, 5% of the pre-existing gingival recession had improved. Thin gingival biotype, visual plaque, and inflammation are useful predictors of gingival recession. PMID- 15877036 TI - Augmentation of faciolingual gingival dimensions with free connective tissue grafts before labial orthodontic tooth movement: an experimental study with a canine model. AB - PURPOSE: The aim of this study was to determine in an animal model whether any increase in gingival thickness after placement of free connective tissue autografts is maintained after labial orthodontic tooth movement. MATERIAL: In a split-mouth technique, the maxillary second and third incisors of 4 adult greyhounds were used as experimental, control, and sham-control teeth. The experimental teeth underwent gingival augmentation surgery. Two to 3 months later, the teeth were moved with a bonded orthodontic appliance over 2 to 3 months. Baseline and 2-month retention clinical measurements of gingival height were taken before the animals were killed. Histometric measurements recorded the free gingival height and the gingival thickness at 5 graduated levels down the teeth. RESULTS: Clinical measurements of changes in gingival margin position revealed a mean coronal shift of 0.44 mm over grafted teeth, with 50% of nongrafted teeth and 100% of the sham-control teeth experiencing small amounts of gingival recession. The histometric results showed that gingival thickness measurements for the grafted teeth were, on average, between 0.13 and 0.18 mm thicker at all levels of measurement than for the nongrafted teeth (P < .01). CONCLUSIONS: Free connective tissue grafts placed on the labial aspect of incisors might help prevent the faciolingual thinning of the gingival tissues that can occur as a result of labial orthodontic tooth movement. Further research is required to confirm these results in a larger sample and determine the long term benefits of preorthodontic gingival augmentation to prevent gingival recession. PMID- 15877037 TI - Effect of self-etching primer and adhesive formulations on the shear bond strength of orthodontic brackets. AB - PURPOSE: Despite many published articles on the bond strength of self-etching primers and adhesives in the restorative dentistry literature, there have been relatively few laboratory studies of the bond strength of new orthodontic materials, and, in most of these published studies, investigators used various methodologic approaches during different stages of the in vitro testing procedures. The aim of this study was to compare the shear bond strength of 4 self-etching primer and adhesive formulations, a nonrinse conditioner and acetone adhesive system, and a conventional system. MATERIAL: The self-etching products tested were Prompt L-Pop (3M ESPE, Seefeld, Germany), Clearfil SE Bond (Kuraray Dental, Osaka, Japan), FL Bond (Shofu Dental, Kyoto, Japan), and One-Up Bond F (Tokuyama, Tokyo, Japan); the nonrinse conditioner and acetone-based adhesive system was NRC and Prime&Bond NT (Dentsply International, Konstanz, Germany); the conventional acid-etch and bond system was Transbond XT (3M Unitek, Monrovia, Calif). Brackets were bonded to intact bovine mandibular incisors (n = 7 per group) according to each manufacturer's recommendations. The specimens were first stored in deionized water at 37 degrees C for 24 hours and then subjected to thermal cycling in deionized water at 5 degrees C +/- 2 degrees C to 55 degrees C +/- 2 degrees C for 1000 cycles. To facilitate degradation of bonds, the specimens were further stored in distilled water for 6 weeks before debonding procedures. RESULTS: The shear bond strengths of the 5 experimental groups were all significantly lower (P < .05) than that of the control group (Prompt L-Pop, 1.72 +/- 0.13 MPa; Clearfil SE Bond, 1.75 +/- 0.19 MPa; FL Bond, 1.71 +/- 0.22 MPa; One-Up Bond F, 1.77 +/- 0.14 MPa; control, 10.5 +/- 0.86 MPa) but not different from one another (P > .05). CONCLUSIONS: The tested self-etching primer and adhesive systems produced bond strength values much lower than that of the control product. Clinically, these products might not be suitable for orthodontic bracket bonding in terms of the shear bond strength achieved after thermal cycling and water storage. PMID- 15877038 TI - Effect of fluoride application on shear bond strength of brackets bonded with a resin-modified glass-ionomer. AB - PURPOSE: The purpose of this study was to assess the effect of fluoride application at 3 different steps of the bonding process on the shear bond strength and bond failure site of a resin-modified glass ionomer cement. MATERIAL: Sixty stainless steel brackets were bonded to bovine incisors with Fuji Ortho LC (GC Europe, Leuven, Belgium) under 4 different enamel conditions: (1) uncontaminated enamel, (2) enamel precleaned with fluoride-containing prophylaxis paste, (3) 1.1% acidulated phosphate fluoride (APF) gel applied immediately before conditioning, and (4) 1.1% APF gel applied immediately before bonding. After bonding, all samples were stored in distilled water for 24 hours and subsequently tested in shear mode on a universal testing machine. RESULTS: No significant differences were found between groups 1 (uncontaminated enamel) and 2 (fluoride application during prophylaxis). Both groups showed significantly greater shear bond strength values than groups 3 (fluoride application before conditioning) and 4 (fluoride application before bonding). Groups 3 and 4 did not differ significantly. Moreover, no significant differences in debond locations were found among the 4 groups. CONCLUSIONS: Fluoride application during initial prophylaxis does not affect the bond strength values of Fuji Ortho LC, whereas it significantly lowers bond strength values when applied before both conditioning and bonding. PMID- 15877039 TI - A longitudinal 3-dimensional size and shape comparison of untreated Class I and Class II subjects. AB - BACKGROUND: The invention of the Broadbent-Bolton cephalometer in 1925 made possible the collection of 3-dimensional data from biorthogonal plain film head radiographs. The objective of this study was to compare longitudinal changes in the shape and size of craniofacial structures between 16 untreated Class II Division 1 girls and 16 untreated Class I Bolton girls. METHODS: Procrustes analyses were used to compare differences in 30 cephalometric landmarks that were 3-dimensional. The same methods were also used to analyze changes of 4 subsets of landmarks (maxilla, mandible, midface, and cranial vault). Comparisons included shape and size differences between adjacent age groups at ages 6, 11, and 15 in the Class II sample as well as between the Class I and Class II samples at each age. RESULTS: Overall, the craniofacial complex underwent continuous shape change from ages 6 to 15 in both samples. In the Class II sample, the smallest contribution to craniofacial shape change was seen for the mandibular landmarks between ages 6 and 11. Compared with the Class I sample, the Class II sample had (1) a longer facial pattern, (2) the smallest mandibular shape difference at age 6 and the largest at age 15, and (3) more protrusive maxillary landmarks at all ages compared with the Class I sample. The Class II sample also had the largest change in size from ages 11 to 15 (6.5%), whereas the Class I sample showed the greatest size change (10.5%) from ages 6 to 11. CONCLUSIONS: Clinically significant size and shape differences were observed during growth and development between Class II and Class I subjects in this sample. PMID- 15877040 TI - The effectiveness of phase I orthodontic treatment in a Medicaid population. AB - BACKGROUND: The effectiveness of early orthodontic treatment for Medicaid enrolled children in the mixed dentition was assessed and compared with results in a population of private-pay patients. MATERIAL: Pre- and posttreatment casts from 196 subjects treated with interceptive orthodontics in the mixed dentition were evaluated by using the peer assessment rating (PAR) index and the index of complexity, outcome, and need (ICON). Ninety-six of the patients were treated at Odessa Brown Community Clinic in Seattle; their treatment was paid by Medicaid. One hundred private-pay patients were treated at the University of Washington graduate orthodontic clinic. The Medicaid and private-pay populations were comparable with respect to initial severity of malocclusion, as assessed by both indexes. RESULTS: PAR and ICON scores fell by similar amounts in the Medicaid (44.1% and 37.5%, respectively) and private-pay (46.8% and 37.3%, respectively) populations. Thus, the groups exhibited similar degrees of improvement with interceptive orthodontic treatment. According to the PAR and ICON, midline discrepancy, overjet, and esthetics exhibited the greatest improvement. The Medicaid population missed significantly more appointments and had poorer oral hygiene than the private-pay group, but these factors did not appear to worsen the outcomes, as measured by the 2 indexes. An appliance with 2 bands and 4 brackets was the most commonly used, and it produced the most significant treatment effect. CONCLUSIONS: Phase I orthodontic treatment significantly reduces malocclusion severity in Medicaid and private-pay populations. There was no difference in initial severity or final outcome, as assessed by the PAR and ICON, between Medicaid and private-pay populations. The degree of improvement in PAR scores did not appear to be associated with compliance. PMID- 15877041 TI - The effectiveness of computer-aided learning in teaching orthodontics: a review of the literature. AB - INTRODUCTION: The purposes of this review were to evaluate the effectiveness of computer-aided learning (CAL) in orthodontic education, to make evidence-based recommendations for the use of CAL in orthodontics, and to develop guidelines for conducting comparative trials to evaluate CAL as a mode of learning in orthodontic education. METHODS: Medline, the Cochrane Library Database, ERIC, CINAHL, LISA, Psycinfo, and IPA were searched for randomized controlled trials evaluating the effectiveness of CAL in orthodontics. Outcome measures included objectively measured posttest scores on multiple choice, written, or oral tests; performance on a clinical procedure or clinical interview; time spent on CAL programs to learn the material presented; and responses to questionnaires conveying participants' attitudes toward various modes of learning. RESULTS: Four randomized controlled trials comparing CAL with conventional teaching fulfilled the inclusion criteria and met the cutoff quality assessment checklist (QAC) score of > or = 8. Each study was assessed for quality by 2 independent reviewers. The validity and strength of the selected studies were assessed by using a QAC for an educational intervention. CONCLUSIONS: The controlled trials of CAL in orthodontics that met our QAC cutoff score of 8 were split, with 2 showing that CAL enjoyed a significant advantage over conventional teaching, 1 showing no difference, and 1 showing that the conventional tutorial method was better. More high-quality trials evaluating the effectiveness of CAL in orthodontics are needed. CAL programs in orthodontics elicit mostly positive responses and attitudes from students toward learning. PMID- 15877042 TI - The nuts and bolts of hemisection treatment: managing congenitally missing mandibular second premolars. AB - When you treat the congenital absence of mandibular second premolars by extracting the residual deciduous second molar and closing the space, you run the risk of flattening facial fullness, especially in patients with nonextraction features--diminished facial fullness, absence of crowding, and reduced tooth size. This challenge is heightened by the increased likelihood that these features will occur in patients who have agenesis. This article describes a new approach that has been shown to minimize such a loss of anchorage. The technique is called hemisection and involves buccolingual sectioning of the second deciduous molar, followed by removal of its distal half. As necessary, the mesial half is also removed, and space closure can be completed. The advantages, philosophy, and technique are discussed in detail. PMID- 15877043 TI - Correction of a Class II deep overbite skeletal and dental asymmetric malocclusion in an adult patient. PMID- 15877044 TI - Understanding science and evidence-based decision making in orthodontics. PMID- 15877045 TI - From 2-dimensional cephalograms to 3-dimensional computed tomography scans. AB - Computed tomography is entering the orthodontic specialty as a mainstream diagnostic modality. Radiation exposure and cost have decreased significantly, and the diagnostic value is very high compared with traditional radiographic options. However, 3-dimensional data present new challenges and need a different approach from traditional viewing of static images to make the most of the available possibilities. Advances in computer hardware and software now enable interactive display of the data on personal computers, with the ability to selectively view soft or hard tissues from any angle. Transfer functions are used to apply transparency and color. Cephalometric measurements can be taken by digitizing points in 3-dimensional coordinates. Application of 3-dimensional data is expected to increase significantly soon and might eventually replace many conventional orthodontic records that are in use today. PMID- 15877046 TI - AdEasy-based cloning system to generate tropism expanded replicating adenoviruses expressing transgenes late in the viral life cycle. AB - Replicating adenoviral vectors (RAds) hold great promise for the treatment of cancer. Significant therapeutic effects of these vectors do not only rely on tumor targeting but also on efficient release of viral progeny from host cells. Cytotoxic genes expressed late in the adenoviral life cycle can significantly enhance viral release and spreading. Therefore, an adenoviral cloning system that allows easy integration of established tumor targeting techniques together with late expression of transgenes can be a valuable tool for the development of RAds. We expanded the features of the widely used AdEasy adenoviral cloning system toward the production of tropism modified replicating adenoviral vectors that express transgenes late in the viral life cycle. Three vectors (pIRES, pFIBER and pAdEasy-Sce) that facilitate easy manipulation of the adenoviral fiber region were established. Unique BstBI and I-Sce-1 restriction sites facilitate the introduction of retargeting peptides in the fiber HI-loop and of genes of interest in the fiber transcription unit. We validated the system by constructing an E1-positive adenovirus with an RGD motif in the fiber HI-loop and green fluorescent protein (GFP) expressed from the fiber transcription unit (AdDelta24Fiber-rgd-GFP). Additionally, assessment of E1-negative replication deficient vectors confirmed strict dependence upon E1 expression for the expression of transgenes inserted into the fiber transcription unit. This flexible cloning system allows for straightforward construction of tropism expanded replicating adenoviral vectors that express transgenes late in the adenoviral life cycle. PMID- 15877047 TI - Silencing p21(Waf1/Cip1/Sdi1) expression increases gene transduction efficiency in primitive human hematopoietic cells. AB - Adult hematopoietic and other tissue stem cells have highly constrained cell cycling that limits their susceptibility to standard gene therapy vectors, which depend upon chromosomal integration. Using cytokine cocktails to increase transduction efficiency often compromises subsequent stem cell function in vivo. We previously showed that p21(Waf1/Cip1/Sdi1) (p21) mediates stem cell quiescence in vivo and decreasing its expression ex vivo leads to an expansion of stem cell pool in vivo. Here, we report that application of p21 specific siRNA increased the gene transduction efficiency in hematopoietic stem cells while preserving cell multipotentiality. Both types of siRNA, synthesized siRNA and transcribed shRNA, reduced p21 expression in target cells by 85-98%. The effect of RNAi in these cells was transient and the level of p21 mRNA returned to base line 14-28 days after siRNA treatment. This brief interval of reduction, however, was sufficient to increase transduction efficiency to two- to four-fold in cell cultures, and followed by a seven- to eight-fold increase in mice. The RNAi treated, lentivector-transduced CD34+ cells retained multipotentiality as assessed in vitro by colony formation assay and in vivo by NOD/SCID mouse transplantation assay. Reduction of p21 resulted in an increased chromosomal integration of lentivector into target cellular DNA. Taken together, both synthesized and transcribed siRNA knocked down p21 expression in human CD34+ hematopoietic stem/progenitor cells. Silencing p21 expression increased gene transduction efficiency and vector integration while retaining stem cell multipotentiality. Thus, RNAi targeting of p21 is a useful strategy to increase stem cell gene transfer efficiency. Decreasing p21 expression transiently while increasing gene-transfer vector integration may ultimately facilitate clinical applications of gene therapy. PMID- 15877048 TI - In vivo NMR imaging evaluation of efficiency and toxicity of gene electrotransfer in rat muscle. AB - In vivo gene electrotransfer (ET) is a simple method of gene delivery in various tissues relying on the injection of plasmid DNA followed by application of electric pulses. Noninvasive tools are needed to evaluate the ET efficiency and the resulting tissue damages. In this study, we performed ET of rat tibialis muscle after injection of either a plasmid coding for luciferase or a contrast agent (CA) detected by using magnetic resonance imaging (MRI). Plasmid expression and CA intracellular trapped quantity were compared throughout the electric field intensity range 0-300 V/cm. Although the CA trapped quantity reflects only the electropermeabilization step, both measurements were correlated. MRI measurements gave easy access to tridimensional visualization of the labelled zones where the CA has been injected and the applied electric field had a value allowing permeabilization. We also performed MRI measurements of the water transverse relaxation time T2 as an indicator of tissue modification, and tested whether another CA specific for necrosis could be used to detect muscle necrosis at high electric field intensity. In conclusion, MRI measurements may bring multiparametric information upon the efficiency and tissue toxicity of an ET protocol by using a simple and safe CA. PMID- 15877049 TI - An efficient construction of conditionally replicating adenoviruses that target tumor cells with multiple factors. AB - Despite the enormous potential of conditionally replicating adenoviruses (CRAs), the time-consuming and laborious methods required to construct CRAs have hampered both the development of CRAs that can specifically target tumors with multiple factors (m-CRA) and the efficient analysis of diverse candidate CRAs. Here, we present a novel method for efficiently constructing diverse m-CRAs. Elements involving viral replication, therapeutic genes, and adenoviral backbones were separately introduced into three plasmids of P1, P2, and P3, respectively, which comprised different antibiotic resistant genes, different ori, and a single loxP (H) sequence. Independently constructed plasmids were combined at 100% accuracy by transformation with originally prepared Cre and specific antibiotics in specific Escherichia coli; transfection of the resulting P1+2+3 plasmids into 293 cells efficiently generated m-CRAs. Moreover, the simultaneous generation of diverse m-CRAs was achieved at 100% accuracy by handling diverse types of P1+2 and P3. Alternatively, co-transfection of P1+3 and P2 plasmids into Cre expressing 293 cells directly generated m-CRA with therapeutic genes. Thus, our three-plasmid system, which allows unrestricted construction and efficient fusion of individual elements, should expedite the process of generating, modifying, and testing diverse m-CRAs for the development of the ideal m-CRA for tumor therapy. PMID- 15877050 TI - Towards mutation-independent silencing of genes involved in retinal degeneration by RNA interference. AB - More than one hundred different mutations in the gene encoding rhodopsin are associated with a group of retinal degenerations including retinitis pigmentosa, congenital stationary night blindness and retinitis punctata albescens. Given this large heterogeneity of mutations, it would be ideal to develop mutation independent therapies for these diseases. We describe use of RNA interference (RNAi) and specifically short hairpin RNAs (shRNAs) expressed from DNA templates to silence both normal and mutant (P23H) human rhodopsin alleles by 94.34+/-2.17 and 94.9+/-1.9%, respectively, in human embryonic retinoblasts. Degeneracy of the genetic code was used to engineer a codon-exchanged mRNA (cmRNA) that demonstrated complete resistance to silencing by the shRNA. Simulation of autosomal dominant retinitis pigmentosa in cell culture through triple transfection of DNAs expressing a cmRNA, a P23H mRNA and an shRNA revealed shRNA mediated silencing, specifically of P23H rhodopsin by 90.64+/-5.19% and no loss of rhodopsin translation from the cmRNA in those cells. In addition, we present data on two alternative shRNA sequences targeting human rhodopsin. Our results have implications for the treatment of a very large variety of retinal degenerations in a mutation-independent manner. PMID- 15877051 TI - Memo to FDA head: IT upgrade required. PMID- 15877052 TI - Earlier stage biotechs attract partners. PMID- 15877053 TI - Cautious welcome for FDA pharmacogenomics guidance. PMID- 15877054 TI - EPO neem patent revocation revives biopiracy debate. PMID- 15877055 TI - US court case to define EST patentability. PMID- 15877056 TI - Syngenta's gaff embarrasses industry and White House. PMID- 15877058 TI - Profile: Julian Thurston. PMID- 15877059 TI - Mixed results in Q1. PMID- 15877060 TI - Gene therapy: cursed or inching towards credibility? PMID- 15877061 TI - The origins of new drugs. PMID- 15877062 TI - Framing the issues on transgenic forests. PMID- 15877063 TI - Lost in the woods. PMID- 15877064 TI - The human cancer genome project--one more misstep in the war on cancer. PMID- 15877065 TI - Beware the biotech barker. PMID- 15877066 TI - Navigating an ethical patchwork--human gene banks. PMID- 15877067 TI - The challenge to patent law of pure chemical protein synthesis. PMID- 15877069 TI - Therapeutic antibody gene transfer. PMID- 15877070 TI - Bringing amyloid into focus. PMID- 15877071 TI - Reverse engineering gene regulatory networks. PMID- 15877072 TI - Tunable antibodies. PMID- 15877074 TI - Systematic interpretation of genetic interactions using protein networks. AB - Genetic interaction analysis,in which two mutations have a combined effect not exhibited by either mutation alone, is a powerful and widespread tool for establishing functional linkages between genes. In the yeast Saccharomyces cerevisiae, ongoing screens have generated >4,800 such genetic interaction data. We demonstrate that by combining these data with information on protein-protein, prote in-DNA or metabolic networks, it is possible to uncover physical mechanisms behind many of the observed genetic effects. Using a probabilistic model, we found that 1,922 genetic interactions are significantly associated with either between- or within-pathway explanations encoded in the physical networks, covering approximately 40% of known genetic interactions. These models predict new functions for 343 proteins and suggest that between-pathway explanations are better than within-pathway explanations at interpreting genetic interactions identified in systematic screens. This study provides a road map for how genetic and physical interactions can be integrated to reveal pathway organization and function. PMID- 15877077 TI - US biotechnology companies and foreign nationals: the changing dynamics of access to H-1B visas. PMID- 15877075 TI - Baculovirus as versatile vectors for protein expression in insect and mammalian cells. AB - Today, many thousands of recombinant proteins, ranging from cytosolic enzymes to membrane-bound proteins, have been successfully produced in baculovirus-infected insect cells. Yet, in addition to its value in producing recombinant proteins in insect cells and larvae, this viral vector system continues to evolve in new and unexpected ways. This is exemplified by the development of engineered insect cell lines to mimic mammalian cell glycosylation of expressed proteins, baculovirus display strategies and the application of the virus as a mammalian-cell gene delivery vector. Novel vector design and cell engineering approaches will serve to further enhance the value of baculovirus technology. PMID- 15877079 TI - Tumour-suppression activity of the proapoptotic regulator Par4. AB - The proapoptotic protein encoded by Par4 (prostate apoptosis response 4) has been implicated in tumour suppression, particularly in the prostate. We report here that Par4-null mice are prone to develop tumours, both spontaneously and on carcinogenic treatment. The endometrium and prostate of Par4-null mice were particularly sensitive to the development of proliferative lesions. Most (80%) Par4-null females presented endometrial hyperplasia by 9 months of age, and a significant proportion (36%) developed endometrial adenocarcinomas after 1 year of age. Similarly, Par4-null males showed a high incidence of prostate hyperplasia and prostatic intraepithelial neoplasias, and were extraordinarily sensitive to testosterone-induced prostate hyperplasia. Finally, the uterus and prostate of young Par4-null mice have increased levels of the apoptosis inhibitor XIAP (X-chromosome-linked inhibitor of apoptosis), supporting the previously proposed function of Par4 as an inhibitor of the (zeta)PKC (atypical protein kinase)-NF-(kappa)B (nuclear factor-(kappa)B)-XIAP pathway. These data show that Par4 has an important role in tumour suppression, with a particular relevance in the endometrium and prostate. PMID- 15877080 TI - Glutamyl-tRNA mediates a switch in RNA polymerase use during chloroplast biogenesis. AB - Chloroplast genes of higher plants are transcribed by two types of RNA polymerase that are encoded by nuclear (NEP (nuclear-encoded plastid RNA polymerase)) or plastid (PEP (plastid-encoded plastid RNA polymerase)) genomes. NEP is largely responsible for the transcription of housekeeping genes during early chloroplast development. Subsequent light-dependent chloroplast maturation is accompanied by repression of NEP activity and activation of PEP. Here, we show that the plastid encoded transfer RNA for glutamate, the expression of which is dependent on PEP, directly binds to and inhibits the transcriptional activity of NEP in vitro. The plastid tRNA(Glu) thus seems to mediate the switch in RNA polymerase usage from NEP to PEP during chloroplast development. PMID- 15877081 TI - Treatment of hepatocellular carcinoma with adenoviral vector-mediated Flt3 ligand gene therapy. AB - Fms-like tyrosine kinase 3 ligand (Flt3L) plays an important role in development and activation of dendritic cells (DCs) and natural killer cells (NK). It has been shown that administration of either tumor cells transfected in vitro with Flt3L vectors or soluble Flt3L fusion protein in a high dose can enhance host antitumor immunity in animal model systems. In this study, we developed a recombinant defective adenovirus with an insert of gene encoding extracellular domain of mouse Flt3L (Ad-mFlt3L) under control of cytomegalovirus promoter and investigated its biological efficacy in eliciting tumor-specific immune response against hepatocellular carcinoma in mouse hepatoma model. The constructed Ad mFlt3L efficiently infected hepa 1-6 hepatoma cells both in vitro and in vivo, leading to a high production of mFlt3L proteins in association with accumulation of DCsNK cells and lymphocytes in local tumor tissues. Tumor cells infected with Ad-mFlt3L lost tumorigenicity and became more immunogenic in syngeniec animal models. Intratumoral injection of Ad-mFlt3L (10(9) expression-forming unit) x 3 significantly inhibited tumor growth with elicitation of long-lasting antitumor immunity, which is both preventive and curative. The tumor-specific immunity can be partially abrogated by depletion of either CD3+CD4+ T cells or NK cells and can be also re-established in naive animals by adoptive transfer of splenocytes from treated mice. The results suggest that adenovirus-mediated Flt3L gene therapy may provide a useful strategy for treatment of cancers. PMID- 15877082 TI - Cotransfection of dendritic cells with RNA coding for HER-2/neu and 4-1BBL increases the induction of tumor antigen specific cytotoxic T lymphocytes. AB - Ribonucleic acid (RNA) transfection of dendritic cells (DCs) was shown to be highly efficient in eliciting CD8+ and CD4+ T-cell responses. We analyzed whether electroporation of DCs with RNA coding for a tumor-associated antigen (TAA) would elicit antigen-specific effector cytotoxic T lymphocyte (CTL) responses and whether these responses could be modulated by cotransfection with a second specific synthetic RNA. Therefore in vitro generated human monocyte-derived DCs were electroporated with in vitro transcribed RNA (in vitro transcript, IVT) encoding the TAA HER-2/neu. Additionally, these cells were cotransfected with IVT coding for human 4-1BBL. Transfection of DCs with 4-1BBL-IVT did not alter their typical phenotype. However, it increased the expression of the costimulatory molecules CD80 and CD40. Coadministration of HER-2/neu- and 4-1BBL-IVT resulted in an increased specific lysis of target cells by the in vitro induced CTL lines, indicating that 4-1BBL enhances their ability to elicit primary CTL responses. Interestingly, transfection of DCs with 4-1BBL-IVT did not augment their capacity to stimulate allogeneic lymphocyte responses. The here established approach of cotransfection of DCs with tumor-RNA and a second specific IVT could improve and optimize the in vitro manipulation of DCs for the induction of antigen-specific CTL responses. PMID- 15877083 TI - Gene therapy for barrett's esophagus: adenoviral gene transfer in different intestinal models. AB - Adenoviral gene therapy could potentially be used for treatment of patients with a Barrett's esophagus. In order to study the feasibility of this approach it is important to study adenoviral intestinal transduction both in vitro and in vivo. In the present study, we used differentiating Caco-2 cells, closed intestinal loops and a Barrett's esophagus rat model to test transduction of adenoviruses expressing green fluorescent protein. We observed a decreased adenoviral transduction from 18.6 to 2.3% in undifferentiated and differentiated Caco-2 cells, respectively. This could be improved by the use of the mucolytic agent N acetylcysteine (NAC) and the polycation diethylaminoethyl-dextran (DEAE-dextran), which improved transduction in differentiated cells five- and ten-fold, respectively. Also an RGD-retargeted adenovirus showed an improved transduction in differentiated cells. In closed intestinal loops adenoviral transduction was limited and the use of NAC and DEAE-dextran or RGD targeting had little effect. The Barrett's esophagus rat model consisted of an esophagojejunostomy, which results in a Barrett's esophagus and esophageal tumors within 6 months. Adenoviral transduction in this model was limited and mainly localized in the basal layer of normal esophagus and stromal tissue of a Barrett's segment. We conclude that although the adenovirus shows promising results in vitro, the current adenoviral vectors are probably not suitable for patients with Barrett's esophagus. PMID- 15877084 TI - Inefficient killing of quiescent human epithelial cells by replicating adenoviruses: potential implications for their use as oncolytic agents. AB - Cultured primary human cells have been widely used to assess the selectivity of oncolytic viruses as potential anticancer agents. As culture conditions can potentially have a significant impact on virus replication and ultimately cell killing, we evaluated the effects of dl309, a wild-type adenovirus, and dl01 / 07, a conditionally replicating adenovirus mutant, on quiescent and proliferating primary mammary epithelial cells. When primary cells were induced into quiescence, both viruses exhibited similar attenuated cell killing. However, cell killing by dl309 was superior to dl01 / 07 in proliferating primary cells. Analysis of viral effects at the level of entry, E2F activation, DNA replication, and late gene expression indicated that attenuation of dl309 in quiescent cells correlated with decreased expression of viral late genes such as hexon. In contrast, attenuation of dl01 / 07 in quiescent cells correlated with inefficient induction of E2F activity and inability to undergo efficient DNA replication. In proliferating cells, dl309 replicated efficiently, whereas dl01 / 07 still showed attenuated replication. In summary, our results indicate the intrinsic preference of wild-type adenoviruses for killing proliferating cells, which is an attractive feature for using adenoviruses as oncolytic agents. These results also highlight the need for the use of appropriate growth conditions for primary cells in vitro to distinguish subtle differences in cell killing among various oncolytic viruses. PMID- 15877085 TI - Recombinant murine interleukin-12 elicits potent antileukemic immune responses in a murine model of Philadelphia chromosome-positive acute lymphoblastic leukemia. AB - Despite the success of chemotherapy regimens in the treatment of acute lymphoblastic leukemia (ALL), certain subsets of patients have a high rate of induction failure and subsequent relapse. One of these subsets of patients carry a translocation between chromosomes 9 and 22, the so called Philadelphia chromosome (Ph+). The result of this translocation is the fusion oncogene, Bcr Abl, which is uniquely expressed in the leukemia clone, and as such has the potential to initiate antileukemic immune responses against the leukemia blasts. We utilized a murine model of Ph+ ALL to look at the ability of systemic interleukin 12 (IL-12) treatments to initiate antileukemic immune responses, and studied the mechanisms by which it does so. We found that IL-12 was able to eliminate pre-established leukemia, and that this protection was mediated by CD4, CD8, and NK cells in combination. While IL-12 was able to eliminate pre established leukemia, it did not elicit immunologic memory. Consistent with previous work, vaccination with irradiated leukemia cells transduced with immunomodulator genes was able to establish long-term memory, and, when used with IL-12, was able to eradicate pre-existing disease and induce resistance to subsequent leukemia challenge. These studies demonstrate the feasibility of an immunotherapeutic approach towards the treatment of Ph+ ALL. PMID- 15877086 TI - Central corneal thickness and its relationship to the patient's origin. AB - OBJECTIVES: To report central corneal thickness (CCT) measurements on patients requesting refractive surgery and to search for possible associations between CCT and patient's origin, age, sex, and preoperative data. SETTING: Department of Ophthalmology, Soroka University Medical Center, and Enaim Refractive Center, Beer-Sheva, Israel. METHODS: Subjects were patients undergoing preoperative examination at our refractive centre during 2003. Patient's age, sex, preoperative CCT, refractive status, keratometry, and intraocular pressure (IOP) were obtained together with country of origin of the patients and their parents. Main outcome measures were CCT measurements and relationship between CCT and patient's origin and preoperative data. RESULTS: A total of 204 patients were included in the study. Patients of North African origin had statistically significantly thinner corneas than patients of other origins (518.9+/-31.5 vs. 545.4+/-30.4 microm in the right eye and 518.4+/-32.1 vs. 546.3+/-29.7 microm in the left eye; P<0.00001). Patients of North African origin were significantly older than patients of other origins (P=0.028). No differences were found when comparing for sex, intraocular pressure, refractive status, and keratometry. When multivariate analysis was performed patient's origin was the only independently associated factor associated with CCT. CONCLUSIONS: This is the first work in reporting lower CCT in people of North African origin examined in a refractive surgery centre. Studies from similar populations are needed to confirm our results. PMID- 15877087 TI - Spontaneous closure of Nd:YAG posterior capsulotomy in capsular blockage syndrome. PMID- 15877088 TI - Late-onset visual decline following successful treatment of subfoveal choroidal neovascularisation with photodynamic therapy. AB - Photodynamic therapy (PDT) has been shown in large studies to be capable of achieving closure of choroidal neovascularization (CNV), thereby resulting in stabilization of visual acuity. We report a series of four patients with classic CNV treated with PDT with good initial result but subsequent severe visual loss that may be related to a change in the morphology of the subretinal scar. PMID- 15877089 TI - Comparative cohort study of the outcomes of deep lamellar keratoplasty and penetrating keratoplasty for keratoconus. AB - AIM: To compare the outcomes and complications of deep lamellar keratoplasty (DLK) and penetrating keratoplasty (PK) for keratoconus. METHODS: A cohort of 20 consecutive PKs, for keratoconus, was compared with 20 consecutive DLKs, for keratoconus. The PKs were performed between June 2000 and July 2001, the DLKs between October 2001 and October 2002. Surgery was performed by one surgeon. Best corrected visual acuities (BCVA), refraction and complications were recorded at the time of surgery, 6 and 12 months postoperatively. chi(2)-tests were used to compare visual acuity outcomes and independent t-tests in the analysis of astigmatism. RESULTS: Groups were comparable for age, sex, and ethnicity. All PKs were uncomplicated. Two of the DLK group had microperforations of Descemet's membrane. There was no significant difference in the proportion of patients achieving 6/9 or better between the PK and DLK groups (85 vs 78%, P=0.54). PK patients were, however, more likely than the DLKs to achieve 6/6 at 1 year; 70% (14/20) of PKs compared to 22% of (4/18) DLKs (P=0.04). Astigmatism was significantly higher in the PKs compared to the DLKs (P=0.022). There were two cases of graft rejection in the PK group, while none in the DLKs. CONCLUSIONS: This study confirms good visual results from both PK and DLK in keratoconus with similarly high percentages reaching 6/9 BCVA. DLK appears to cause less astigmatism and also has the advantage of no endothelial graft rejection. The apparent cost, however, is a reduction in the likelihood of achieving 6/6 BCVA. PMID- 15877090 TI - Rothia genus endophthalmitis following penetrating injury in a child. PMID- 15877091 TI - Exudative retinal detachment after photodynamic therapy: a case report in an Asian patient. PMID- 15877092 TI - Influence of age on conjunctival bacteria of patients undergoing cataract surgery. AB - AIM: To ascertain the effect of elderly in the conjunctival bacteria frequency of patients undergoing cataract extraction. METHODS: A retrospective case series study of 4432 consecutive patients who underwent cataract surgery, without excluding any of them for having pre-existing diseases. Their preoperative conjunctival culture were performed over a 3-year period (1994-1996). Bacteria were grouped in nine categories and patients were divided into seven groups, according to age; comparisons between groups were made by means of the chi(2) test, and the Mantel-Haenszel test to analyse age as a confounder, using SPSS program, version 12. RESULTS: Patients aged over 74 years accounted for 41.4%; women predominated among this group (61.4%), but not in the patients younger than 75 (47.7%); In the whole sample women accounted for 53.4%. Patients aged 75-96 years had a greater frequency of: Corynebacteria, Staphylococcus aureus, Streptococcus sp. (except Streptococcus pneumoniae), Gram-negative cocci and Gram negative rods (except Haemophilus sp.) and 'other bacteria' categories than those aged 3-74 years. Different bacteria frequency in both sexes produced a confounding effect in the comparison between age-groups. Men had more Staphylococci coagulase (-), S. pneumoniae and Gram-negative rods than women. CONCLUSIONS: Elderly patients awaiting cataract surgery had more conjunctival bacteria than those younger than 75 years, except Staphylococcus coagulase negative, S. pneumoniae and Haemophilus sp. At any age, men had more bacteria than women. These increments of frequencies could increase the risk of intraocular surgery contamination. PMID- 15877093 TI - Corticosteroid-induced glaucoma: a review of the literature. AB - The intraocular pressure rise that can complicate the use of topical or systemic corticosteroid has been recognised for 50 years. More recently, following isolation of the myocilin gene (previously known as the trabecular meshwork inducible glucocorticoid response or TIGR gene), there has been renewed interest in this steroid-responsive phenomenon. Furthermore, the currently fashionable use of injectable intraocular steroids in the management of clinically significant subretinal fluid and macular oedema has resulted in an increased incidence. Animal studies, cell biology, molecular biology, and an improved knowledge of genetics have provided a better understanding of the mechanisms behind the response. The purpose of this review is to describe the risk factors for developing corticosteroid-induced glaucoma, to discuss the underlying mechanisms and genetics of the condition and to present management options. PMID- 15877094 TI - 'Fast-tracking' cancer referrals: application for periocular basal cell carcinoma. AB - AIM: To study the feasibility of a modified fast-track protocol for periocular basal cell carcinoma (BCC). METHODS: A modified protocol was set up with an aim to examine all periocular BCCs within 6 weeks of referral to our oculoplastic clinic. An audit of this protocol was performed over a 2-year period. RESULTS: A total of 65 patients were referred as a 'possible BCC' over the 2-year period. In all, 32 of these patients were referred by dermatologists (49%), followed by fellow consultant ophthalmologists (20 patients, 31%) and general practitioners (11 patients, 17%). The clinical suspicion in the oculoplastic clinic agreed with the referral diagnosis in 71% (46 of the 65) of patients. This figure was particularly high for referrals from dermatologists (84%). In all, 44 out of the 46 'clinically suspected BCC' underwent surgical excision. Histopathology confirmed BCC in 39 of these 44 patients, a diagnostic accuracy of 89%. Among the subgroup of patients referred by the dermatologists, the largest source of referrals, 24 out of 30 patients that underwent surgical excision had histologically proven BCC; a diagnostic accuracy of 80%. CONCLUSION: Our study shows that the modified fast-track protocol for periocular BCCs is practical and feasible. Such a practice is highly desirable since it prevents a long wait for patients who are aware of a possible malignant periocular lesion. PMID- 15877095 TI - Modification of diabetic retinopathy presentation by underlying hepatitis C infection. PMID- 15877096 TI - Delayed retrobulbar haemorrhage after Ahmed glaucoma implant: a case report. PMID- 15877097 TI - Nonpreserved human amniotic membrane transplantation for conjunctival reconstruction after excision of extensive ocular surface neoplasia. AB - PURPOSE: To report our experience on the use of nonpreserved human amniotic membrane transplantation (AMT) in ocular surface reconstruction after excision of extensive ocular surface neoplasia (OSN). DESIGN: Prospective noncomparative interventional case series. PARTICIPANTS: In all, 10 eyes of 10 consecutive patients with extensive OSN involving various areas of limbus, conjunctiva, and cornea (conjunctival carcinoma in situ, four eyes; squamous cell carcinoma, three eyes; malignant melanoma, two eyes; conjunctival-orbital lymphangioma, one eye) were included in this prospective noncomparative interventional case series. After excision of the neoplasia with 3-4 mm tumour-free margins, double freeze thaw cryotherapy was applied to the margins of the remaining conjunctiva, and nonpreserved human amniotic membrane graft was sutured to the adjacent conjunctiva using 8/0 vicryl sutures and cornea using 10/0 nylon sutures, with the epithelial side facing up to cover the bare sclera and cornea. Postoperatively, topical corticosteroids were used for 3 months. RESULTS: After tumour excision and AMT, a satisfactory result with a wet, stable conjunctiva, and rapid and complete healing was observed in all eyes. Over a mean follow-up of 10.0 months (range, 6-27 months), all but one eye remained free of tumour recurrence. In one eye with conjunctival melanoma, there was a small recurrence, which was treated with excision and cryotherapy. Treatment complications were partial stem cell deficiency in two eyes and symblepharon formation in one eye. Immune graft rejection was not encountered. CONCLUSION: Nonpreserved human AMT appears to be useful for reconstruction of ocular surface following excision of extensive OSN. PMID- 15877099 TI - Precautions in ophthalmic practice in a hospital with a major acute SARS outbreak: an experience from Hong Kong. AB - Many new infectious diseases in humans have been derived from animal sources in the past 20 years. Some are highly contagious and fatal. Vaccination may not be available and antiviral drugs are not effective enough. Infectious control is important in clinical medicine and in Ophthalmology. Severe acute respiratory syndrome (SARS), as an example, is a highly contagious respiratory disease that has recently been reported in Asia, North America, and Europe. Within a matter of weeks, the outbreak has evolved to become a global health threat and more than 30 countries have been afflicted with a novel Coronavirus strain (SARS-CoV) that is the aetiologic agent of SARS. The primary route of transmission of SARS appears involving close person-to-person contact through droplets. Ophthalmologists may be particularly susceptible to the infection as routine ophthalmic examinations like direct ophthalmoscopy and slit-lamp examination are usually performed in a setting that has close doctor-patient contact. Being the Ophthalmology Department of the only hospital in the world that has just gone through the largest outbreak of SARS, we would like to share our strategy, measures, and experiences of preventing contracting or spreading of SARS infection as an infection control model. SARS is one of the many viruses against which personnel will need protecting in an ophthalmic setting. The experiences attained and the measures established might also apply to other infectious conditions spreading by droplets such as the avian influenza with H5N1. PMID- 15877100 TI - Acute irreversible diabetic cataract in adolescence: a case report. PMID- 15877101 TI - Diode laser transcleral cyclophotocoagulation for refractory glaucoma: a 1 year follow-up of patients treated using an aggressive protocol. AB - PURPOSE: To prospectively evaluate the intraocular pressure (IOP) lowering ability, retreatment rate, and complications of transcleral Diode laser cyclophotocoagulation using a higher power setting than what is generally recommended. PATIENTS AND METHODS: A total of 36 eyes of thirty six patients with refractory glaucoma, and who fitted our inclusion criteria underwent Diode cyclophotocoagulation. The laser power was set at 2250 mW, with a duration of 2000 ms, and a total number of 28 shots for the first treatment and 20 shots for any consequent one. The patients were followed up for 1 year with the following outcomes being analysed: IOP, visual acuity, change in the number of medications, and complications. RESULTS: The mean IOP decrease was 53% (P < 0.05), and 72.2% of the patients maintained an IOP < or =21 mmHg for the whole duration of the study The number of medications necessary to control the pressure, significantly dropped from 2.8 to 0.89 (P < 0.05), and 25% of the patients needed the treatment to be repeated only once. In all, 33% of the patients improved their visual acuity after the treatment, while 22% worsened, and the rest stayed the same. The most common treatment complications were conjunctival injection and corneal oedema, and these were both transient and reversible. CONCLUSION: The use of the higher power setting of 2250 mW, resulted in a sustained lower IOP, less use of medications, less need for retreatment, relative preservation of visual acuity, and only reversible complications. PMID- 15877102 TI - A comparative study of epiretinal membranes associated with Eales' disease: a clinicopathologic evaluation. AB - AIM: To study the histopathologic features and clinical correlation of epiretinal membranes (ERM) obtained from patients of Eales' disease and compare with other vasoproliferative disorders. METHODS: Retrospective analysis of epiretinal membranes submitted for histological evaluation between January 1995 and June 2001, from the patients of diabetic retinopathy and vascular occlusions (Group 1; vaso-occlusive disorders) and of Eales' disease (Group 2; vasoinflammatory disorders). Demographics, pre and postoperative visual acuity, and anatomic and histologic characteristics of membranes were studied. Histopathologic features and clinical outcomes were correlated between the groups. The results were analysed statistically by Student's t-test, Fisher's exact test and Kruskal Wallis test. RESULTS: This study consisted of 42 patients, 24 in Group 1 and 18 in Group 2. Patients in Group 2 (33.0+/-9.2 years) were significantly younger than the patients in Group 1 (49.9+/-7.6 years) (P< or =0.0001). Final visual acuity of >20/400 was attained in 79.2% (19/24) patients in Group 1 and 83.3% (15/18) in Group 2 (P=1.0). Inflammatory membranes were significantly associated with presumed Eales' disease (94.4 vs 0%) (P< or =0.0001) and fibrovascular membranes with Group 1 (70.8% vs 33.3%) (P=0.028). Mast cells and eosinophils were observed as special features in epiretinal membranes of patients with Eales' disease. CONCLUSIONS: Histological features of ERM in Eales' disease are comparable to other vasoproliferative disorders except for features of inflammation. Presence of mast cells and eosinophils in epiretinal membranes of Eales' disease needs further investigation. PMID- 15877103 TI - National Biometry Audit II. AB - PURPOSE: To determine the change in compliance with the Royal College of Ophthalmologists biometry guidelines since the last National Audit 2 years ago and in particular to quantify the adoption of modern methods of axial length measurement and customization of A constants. METHOD: A structured telephone questionnaire of individuals who perform biometry in all eye departments in the United Kingdom. RESULTS: A biometrist was interviewed in 94 of the 178 United Kingdom Ophthalmology departments. Compared with 2 years ago, nurses alone perform biometry more frequently (67 vs 51%) and junior doctors less frequently (9 vs 15%). More biometrists now attend external training courses (45 vs 37%). The Royal College of Ophthalmologists recommended intraocular lens calculation formulae (SRK-T, Hoffer Q, and Holladay) are used more commonly (30 and 15%) and audit of prediction error is being performed more frequently (78 vs 71%). The routine use of a partial coherence laser interferometry has increased from 35 to 61% in United Kingdom Ophthalmology departments. Currently, only one United Kingdom department is routinely using immersion ultrasound biometry. 'A' constants are customized in 47% of departments. CONCLUSION: Over the last 2 years, there has been improved implementation of the Royal College of Ophthalmologists guidelines on biometry. It is essential that the Royal College of Ophthalmologists guidelines are updated to include current best practice of routine use of partial coherence laser interferometry or immersion biometry and customization of A constants. A benchmark standard of 85-90% of patients achieving a final postoperative refraction within 1 dioptre of the predicted should be established. PMID- 15877104 TI - Outcome of cataract extraction and posterior chamber intraocular lens implantation following glaucoma filtration surgery. AB - PURPOSE: To evaluate the outcome of cataract extraction (CE) after glaucoma filtering surgery (GFS). METHODS: A total of 77 eyes (77 patients) who underwent CE with posterior chamber intraocular lens (PCIOL) implantation following GFS by a single surgeon were reviewed. Main outcome measures were preoperative and postoperative intraocular pressures (IOPs), visual acuities, medications, astigmatism, bleb survival, time of surgical failure, complications, and success rate. RESULTS: Mean time interval between GFS and CE was 46.8+/-50.9 months (range, 2-348 months). The mean preoperative IOP was 13.9+/-4.7 mmHg (range 3-27 mmHg) and mean postoperative IOP at 3 weeks was 13.6+/-5.5 mmHg (range, 6-44 mmHg). The mean follow-up was 19.5+/-20.1 months (range, 1.4-73 months; median 10.6 months). Complete success was achieved in 59 eyes (76.7%). The cumulative probability of complete success was 91.3+/-3.7, 82.0+/-5.6 and 78.1+/-6.5% at the end of 6 months, 1, and 2 years, respectively. Visual acuity before CE was < or =20/50 in all eyes (100%). Visual acuity at last visit was > or =20/40 in 33 eyes (42.8 %), 20/50-20/80 in 30 eyes (39.0%), < or =20/100 in 14 eyes (18.2%). Risk factors identified for qualified success included age at CE>60 years, interval of < or =5 months between GFS and CE, use of preoperative glaucoma medications, and postoperative IOP >19 mmHg within 2 weeks. CONCLUSIONS: IOP and bleb function was maintained after CE with PCIOL implantation following successful GFS with good visual recovery. PMID- 15877105 TI - Decreased susceptibility of differentiated PC12 cells to oxidative challenge: relationship to cellular redox and expression of apoptotic protease activator factor-1. AB - We previously showed that tert-butyl hydroperoxide (TBH) induced apoptosis in naive rat pheochromocytoma (nPC12) cells that correlated with cellular redox imbalance and mitochondrial apoptotic signaling. In this study, we tested the hypothesis that differentiation of nPC12 cells results in altered susceptibility to TBH utilizing a model of differentiated PC12 (dPC12) cells induced by nerve growth factor. TBH (100 microM) induced dPC12 apoptosis (12% at 24 h) at levels lower than naive cells (35%). This resistance was associated with elevated GSH, NADPH (reduced nicotinamide adenine dinucleotide phosphate), TBH metabolism, redox enzyme activities, reduced cellular GSH/GSSG (glutathione disulfide) status and preservation of mitochondrial membrane potential. Altering cellular GSH with ethacrynic acid or N-acetylcysteine, respectively, exacerbated or protected against dPC12 apoptosis. dPC12 apoptosis was mediated by caspase-9 and -3 activation and apoptosis protease activator protein-1 (Apaf-1) expression. These results show that nPC12 transition to dPC12 cells afforded protection against oxidative challenge due to maintenance of reduced GSH/GSSG and decreased Apaf-1 expression. PMID- 15877106 TI - Effects of inducible overexpression of DNp73alpha on cancer cell growth and response to treatment in vitro and in vivo. AB - The p73 gene has a complex regulation, which leads to the expression of different isoforms, often with opposite biological effects. We have generated in the human colocarcinoma cell line HCT116, expressing a wild-type p53, an inducible DNp73alpha expressing system. Two clones (HCT116/DN3 and HCT116/DN14), upon doxycycline addition, show a strong expression of DNp73alpha. In vitro the two DNp73alpha overexpressing clones grow at similar rate of the control transfected clone (HCT116/8a) and similarly respond to DNA damage. When injected in mice, HCT116/DN3, HCT116/DN14, and HCT116/8a cells grew similarly in the absence or presence of tetracycline. In HCT116/DN3 and HCT116/DN14 tumors, tetracycline induced a strong expression of DNp73alpha both as mRNA and protein. These results indicate that in this system the overexpression of the DNp73alpha does not induce a more aggressive phenotype and does not seem to be associated with a reduced response of the cells to treatment with anticancer agents. PMID- 15877107 TI - Emerging issues in traditional Chinese medicine. AB - Traditional Chinese medicine (TCM) has many beneficial effects and has been practiced for several thousand years. It is known to treat the cause of a disease rather than to alleviate its symptoms. Based on a belief that TCM is natural, safe, and of lower cost, consumers worldwide are spending more out-of-pocket money on this form of therapy. This increased spending, and reports of adverse reactions, has drawn the attention of many regulatory agencies. Scientists have called for more evidence-based and scientific research on the risks and benefits of TCM. In Canada, the Natural Health Product Regulations came into effect January 2004. TCM herbal product manufacturers will need to provide products of reputable quality to the market. Many will apply modern technology and good science to support their products. The issues facing producers, scientists, and consumers alike are quality control and assessment, standardization of bioactive components, mechanisms of actions, and integration of the evolved modern Chinese medicine into the healthcare system. Solid science, better regulation of the final product, and better education of consumers are necessary to extract the best of TCM to complement existing conventional medicine to deliver the best healthcare. PMID- 15877108 TI - Alpha 1 adrenergic receptor control of renal blood vessels during aging. AB - Aging humans and rats have a reduced renal vascular constriction response to stress, change in posture, or exercise. In this study, renal interlobar arteries from 9- (intermediate age) to 15-month-old (aging) male Wistar rats constricted less to alpha-adrenergic agonists than those of 4-month-old (young adult) rats. The reduced contraction to A61603 (alpha 1 A agonist) was similar to that to norepinephrine and phenylephrine. Therefore, it appears that the reduction in constriction is primarily related to alpha 1 A receptor stimulation. GeneChip microarray hybridization analysis of the interlobar arteries with the RAE 230A GeneChip indicated that there were no significant differences in gene expression for alpha 1 A/C, 1B, or 1D receptors between 4-month-old (young adult) and 1-year old (aging) male Wistar rats. Competitive binding experiments (prazosin) revealed that maximal binding (Bmax, fmol/mg protein) of the alpha 1 receptors of interlobar arteries was reduced 25% by 10 months of age and 50% by 18+ months of age. Alpha 1 receptor-induced arterial constriction and prazosin binding were both down-regulated. The loss of receptor-initiated constriction likely includes down-regulation of maximum agonist binding by alpha 1 adrenergic receptors. PMID- 15877109 TI - Partial reversal by rutin and quercetin of impaired cardiac function in streptozotocin-induced diabetic rats. AB - The present investigation was carried out to evaluate the effects of the cyclodextrin complexes quercetin and rutin on left ventricle dysfunction in streptozotocin-induced diabetic rats. Diabetes was induced by streptozotocin (45 mg/kg body mass, i.v.) in Sprague-Dawley rats. Echocardiography and biochemical and histological studies were carried out under normal control, diabetic untreated, normal and diabetic vehicle (beta-cyclodextrin, p.o.), quercetin- (100 and 300 mg/kg, p.o.), and rutin- (100 and 300 mg/kg, p.o.) treated normal and diabetic animals at varying time intervals (1 and 12 weeks). The increase in the serum triglycerides and cholesterol levels was attenuated in the cyclo dextrin complexes of rutin-treated animals significantly more than in the quercetin treated and diabetic vehicle-treated animals. Left ventricular diastolic dysfunction was observed in diabetic vehicle-treated animals after 12 weeks of the study as determined by a significant decrease in E-wave (45.91%), an increase in the A-wave (75.55%), and a decrease in the E/A ratio (70.14%). However, the percent decrease (after 12 weeks) in the E-wave, increase in the A-wave, and decrease in the E/A ratio were less in the cyclodextrin complexes of rutin treated animals (100 and 300 mg/kg), which had the following values: E-wave, 12.22% and 13.80%; A-wave, 25.90% and 10.40%; and E/A ratio, 31.01% and 20.52%. In the quercetin-treated animals (100 and 300 mg/kg), which had the following values: E-wave, 40.44% and 36.44%; A-wave, 52.98% and 29.28%; and E/A ratio, 61.70% and 51.11%. Histopathological studies revealed that the degree of myocardial necrosis was less in rutin-treated animals compared with quercetin and diabetic vehicle-treated animals: rutin < quercetin < beta-cyclodextrin. Myocardial fructose levels were significantly increased in the diabetic vehicle treated animals after 12 weeks of the study, suggesting an increment in the myocardial polyol pathway activity. However, myocardial fructose levels were significantly decreased in the rutin- and quercetin-treated animals compared with the vehicle-treated animals, possibly owing to their aldose reductase inhibitory activity. Quercetin and rutin treatment did not influence the echocardiographical and histo logical parameters in normal animals. Results from the present investigation demonstrated that rutin has a cardioprotective activity, and we conclude that the observed cardioprotection with rutin may be due to its aldose reductase inhibitory activity, as the enhanced aldose reductase pathway is implicated in the development of left ventricle dysfunction by several studies. PMID- 15877110 TI - Modification of sarcoplasmic reticulum (SR) Ca2+ release by FK506 induces defective excitation-contraction coupling only when SR Ca2+ recycling is disturbed. AB - This study examined whether the effects of FK506-binding protein dissociation from sarcoplasmic reticulum (SR) Ca(2+) release channels on excitation contraction (EC) coupling changed when SR Ca(2+) reuptake and (or) the trans sarcolemmal Ca(2+) extrusion were altered. The steady-state twitch Ca(2+) transient (CaT), cell shortening, post-rest caffeine-induced CaT, and Ca(2+) sparks were measured in rat ventricular myocytes using laser-scanning confocal microscopy. In the normal condition, 50 micromol FK506/L significantly increased steady-state CaT, cell shortening, and post-rest caffeine-induced CaT. When the cells were solely perfused with thapsigargin, FK506 did not reduce any of the states, but when low [Ca(2+)](0) (0.1 mmol/L) was perfused additionally, FK506 reduced CaT and cell shortening, and accelerated the reduction of post-rest caffeine-induced CaT. FK506 significantly increased Ca(2+) spark frequency in the normal condition, whereas it mainly prolonged duration of individual Ca(2+) sparks under the combination of thapsigargin and low [Ca(2+)](0) perfusion. Modification of SR Ca(2+) release by FK506 impaired EC coupling only when released Ca(2+) could not be taken back into the SR and was readily extruded to the extracellular space. Our findings could partly explain the controversy regarding the contribution of FK506-binding protein dissociation to defective EC coupling. PMID- 15877111 TI - Effects of sodium fluoride on the mechanical activity in mouse gastric preparations. AB - The aim of the present study was to investigate the responses induced by sodium fluoride (NaF) on gastric mechanical activity, using mouse whole-stomach preparations. The mechanical activity was recorded in vitro as changes of intraluminal pressure. In most of the preparations, NaF induced a tetrodotoxin insensitive biphasic effect characterized by early relaxation followed by slowly developing contractile response. The contraction was dependent on the concentration of NaF, whereas the relaxation was observed at only 10-30 mmol/L NaF. The contractile effect was significantly reduced by nifedipine (an L-type Ca(2+) channel blocker), ryanodine or ruthenium red (inhibitors of Ca(2+) release from sarcoplasmic reticulum), and GF109203X (a protein kinase C inhibitor). Moreover, it was abolished by neomycin (an inhibitor of phospholipase C) and potentiated by SQ22536 (an inhibitor of adenylyl cyclase). All the drugs significantly increased the relaxation, except SQ22536, which abolished it. The present results suggest that NaF causes a complex mechanical response in the whole-stomach, which might explain gastric discomfort after fluoride ingestion. The relaxation appears owing to production of cAMP, while the contractile effects imply activation of phospholipase C, protein kinase C, influx of Ca(2+), and release of Ca(2+) from ryanodine-sensitive intracellular store. PMID- 15877112 TI - Acetylcholine-induced phosphorylation of CPI-17 in rat bronchial smooth muscle: the roles of Rho-kinase and protein kinase C. AB - It has been demonstrated that CPI-17 provokes an inhibition of myosin light chain phosphatase to increase myosin light chain phosphorylaton and Ca(2+) sensitivity during contraction of vascular smooth muscle. However, expression and agonist mediated regulation of CPI-17 in bronchial smooth muscle have not been documented. Thus, expression and phosphorylation of CPI-17 mediated by PKC and ROCK were investigated using rat bronchial preparations. Acetylcholine (ACh) induced contraction and Ca(2+) sensitization were both attenuated by 10(-6) mol Y 27632 /L, a ROCK inhibitor, 10(-6) mol calphostin C/L, a PKC inhibitor, and their combination. A PKC activator, PDBu, induced a Ca(2+) sensitization in alpha-toxin permeabilized bronchial smooth muscle. In this case, the Ca(2+) sensitizing effect was significantly inhibited by caphostin C but not by Y-27632. An immunoblot study demonstrated CPI-17 expression in the rat bronchial smooth muscle. Acetylcholine induced a phosphorylation of CPI-17 in a concentration dependent manner, which was significantly inhibited by Y-27632 and calphostin C. In conclusion, these data suggest that both PKC and ROCK are involved in force development, Ca(2+) sensitization, and CPI-17 phosphorylation induced by ACh stimulation in rat bronchial smooth muscle. As such, RhoA/ROCK, PKC/CPI-17, and RhoA/ROCK/CPI pathways may play important roles in the ACh-induced Ca(2+) sensitization of bronchial smooth muscle contraction. PMID- 15877113 TI - Increased incidence of hepatic insulin-sensitizing substance (HISS)-dependent insulin resistance in female rats prenatally exposed to ethanol. AB - Insulin causes the release of the hepatic insulin-sensitizing substance (HISS) from the liver. Hepatic parasympathetic nerves play a permissive role in the release of HISS. HISS-dependent insulin resistance (HDIR) occurs in the absence of HISS. Fetal ethanol exposure has been shown to cause dose-dependent HDIR in adult male rat offspring. Since female offspring are more severely affected by in utero ethanol toxicity, we hypothesized that fetal alcohol exposure causes higher incidence and more severe HDIR in adult female offspring. Adult female rat offspring prenatally exposed to different concentrations of ethanol (0%, 15%, and 20%) were tested for insulin sensitivity using the rapid insulin sensitivity test (RIST). The RIST index was significantly reduced in the 15% (134.1 +/- 16.1 mg/kg) and the 20% (98.7 +/- 9.7 mg/kg) group compared with the 0% (220.9 +/- 27.6 mg/kg) group. Administration of atropine produced significant additional HDIR in the 15% group (82.9 +/- 14.5 mg/kg) but not the 20% group (83.8 +/- 20.5 mg/kg) indicating complete HDIR had been produced in this group, contrary to the adult male offspring in a previous study. The results are consistent with the hypothesis that adult-female offspring are more severely affected by in utero ethanol exposure compared with adult-male offspring. PMID- 15877114 TI - Alkyne metathesis. AB - This review discusses the emergence of alkyne metathesis as a valuable synthetic tool applicable in the synthesis of complex molecules and polymer science. PMID- 15877115 TI - A new self-assembling capsule via metal coordination. AB - A new octadentate cavitand forms a stable dimeric molecular capsule via metal coordination, creating a large and elaborate three-dimensional cavity in which large aromatic guests are accommodated to form supramolecular complexes. PMID- 15877116 TI - Cyclic dimer of a fused porphyrin zinc complex as a novel host with two pi electronically coupled binding sites. AB - Upon complexation with 4,4[prime or minute]-bipyridine, a cyclic dimer of a fused porphyrin zinc complex, having two pi-electronically coupled binding sites, shows a strong negative cooperativity in the second guest binding, to allow stepwise formation of 1 : 1 and 1 : 2 inclusion complexes. PMID- 15877117 TI - A two-dimensional cationic lattice built from [Zn6(HPO4)2(PO4)2]2+ clusters. AB - A unique cationic zinc phosphate cluster linked by neutral bifunctional rigid ligands to form a two dimensional framework was synthesized and structurally characterized. PMID- 15877118 TI - Theoretical quest for the titanium-substituted hydrocarbons. AB - Using theoretical calculations we predict a novel family of compounds, which might serve for hydrogen storage, and for chemical vapour deposition of titanium carbide coatings. PMID- 15877119 TI - Supramolecular isomers in the same crystal: a new type of entanglement involving ribbons of rings and 2D (4,4) networks polycatenated in a 3D architecture. AB - The reaction of corresponding cadmium salts with the flexible ligand 1,4 bis(1,2,4-triazol-1-ylmethyl)benzene (bbtz) affords the 3D coordination network [Cd3(bbtz)6(H2O)6](BF4)(6.1.75H2O (1), containing ribbons of rings and planar 2D (4,4) networks polycatenated with each other, the undulating 2D (4,4) network [Cd(bbtz)2(H2O)2](ClO4)2.2H2O (2), and ribbons of rings [Cd(bbtz)2(H2O)2](BF4)2.3DMF (3) and [Cd(bbtz)2(H2O)2](ClO4)2.3DMF (4). PMID- 15877120 TI - Observation of an octameric water cluster containing a book-shaped hexamer in a 4f-3d complex. AB - An octameric water cluster consisting of a book-shaped water hexamer and two dangling water molecules has been observed in a 4f-3d ionic pair complex [Y(dpdo)2(H2O)4][Co(CN)6].4H2O. PMID- 15877121 TI - Structure and magnetic properties of a novel copper halide framework {[tBuNH3]2[Cu3(mu3-OH)(mu2-H2O)Cl7]}n synthesized via in situ templation. AB - The use of the tris(alkylamido)phosphate OP[N(H)tBu]3 as an in situ source of the templating agent [tBuNH3]+ produces the copper halide chain {[tBuNH3]2[Cu3(mu3 OH)(mu2-H2O)Cl7]}n (1) in a solvothermal process; the novel antiferromagnetically coupled trinuclear fragment Cu3(OH)(H2O)Cl7 is the building block in the polymeric anion in . PMID- 15877122 TI - Selective O2 oxidation of air-sensitive lanthanocene thiolates and thioether chelate. AB - Two unusual regioselective O2 oxidation reactions of air-sensitive lanthanocene thiolates and a thioether chelate are described, revealing a novel oxygenation pattern of thiolate ligands. PMID- 15877123 TI - Enantioselective allylation of aldehydes catalyzed by chiral indium(III) complexes immobilized in ionic liquids. AB - In the presence of chiral catalytic complexes prepared from In(OTf)3 and chiral PYBOX ligands, allytributylstannane reacted with aldehydes in ionic liquids to afford the corresponding homoallylic alcohols in high enantioselectivities (86 94% ee) and good yields (68-89%); the chiral catalysts immobilized in ionic liquids could be reused with comparable enantioselectivities and yields. PMID- 15877124 TI - Penicillin G acylase catalyzed Markovnikov addition of allopurinol to vinyl ester. AB - A new enzymatic process is reported, in which penicillin G acylase from Escherichia coli displays a promiscuous activity in catalyzing the Markovnikov addition of allopurinol to vinyl ester. PMID- 15877125 TI - Unexpected promotion of Au/TiO2 by nitrate for CO oxidation. AB - The catalytic activity for Au/TiO2 for CO oxidation can be significantly enhanced by the addition of nitrates and this may relate to the variable catalyst performance observed in many studies. PMID- 15877126 TI - Phosphonate mediated surface reaction and reorganization: implications for the mechanism controlling cement hydration inhibition. AB - Vertical scanning interferometry and XPS show the reaction of CaCO3 with the hydration retarder nitrilo-tris-(methylene)phosphonic acid follows a pathway of dissolution of the calcium followed by precipitation of a calcium phosphonate; subsequent surface reorganization/restructuring of the calcium phosphonate exposes the underlying CaCO3 for further hydration. PMID- 15877127 TI - Pentacoordinated germanium in AST zeolite synthesised in fluoride media. A 19F NMR validated computational study. AB - A computational study shows that Ge is pentacoordinated in the double four rings (D4R) of Si/Ge AST zeolites; the calculated chemical shifts of F-D4R containing 8Si, 7Si1Ge and 8Ge reproduce the trends of 19F NMR experiments. PMID- 15877128 TI - Direct observation of a cooperative mechanism in the adsorption of heavy metal ions to thiolated surface by in-situ surface plasmon resonance measurements. AB - S-type adsorption kinetics, obtained by surface plasmon resonance spectroscopy measurements, suggest that the rate of Pt2+ adsorption on 1,6-hexanedithiol (HDT) on gold increases until the surface coverage reaches ca. 17%, after which, the adsorption profile of Pt2+ follows Langmuirian behavior for the surface coverage. PMID- 15877129 TI - Novel mesoporous silica-perfluorosulfonic acid hybrids as strong heterogeneous Bronsted catalysts. AB - Perfluorinated sulfonic acids have been immobilised into mesoporous silica frameworks by a one-step process, representing the first example of the successful incorporation of charged silanes using this route, and have been shown to be excellent catalysts for Bronsted acid catalysed transformations. PMID- 15877130 TI - Evolution of the pseudo-1,3-dipolar cycloaddition chemistry of SNSMF6 (M = As, Sb) leading to 2,5-dihydroxybenzo-1,3,2-dithiazolylium and 2,7-dicarbonylnaphtha 1,3,2-dithiazolylium salts and their corresponding radicals. AB - We report the unprecedented formation of a benzo-fused 1,3,2-dithiazolylium [AsF6 ] salt by a one step, quantitative, cycloaddition of SNSAsF6 with 1,4 benzoquinone. In contrast, the reaction of SNSSbF6 with 1,4-naphthaquinone results in 2,7-dicarbonylnaphtha-1,3,2-dithiazolylium [SbF6-]. Both were reduced to the corresponding 7pi radicals. PMID- 15877131 TI - Lipase-catalyzed domino kinetic resolution of alpha hydroxynitrones/intramolecular 1,3-dipolar cycloaddition: a concise asymmetric total synthesis of (-)-rosmarinecine. AB - The title domino reactions were developed to directly provide tetrahydrofuro[3,4 c]isoxazole derivatives (5 and 9) in > or =90% ee from racemic alpha hydroxynitrones (2 and 6), which were used in the concise asymmetric total synthesis of (-)-rosmarinecine . PMID- 15877132 TI - A novel strategy for the asymmetric synthesis of chiral cyclopropane carboxaldehydes. AB - A new way of combining chiral auxiliaries and substrate-directable reactions for asymmetric synthesis is described that employs a three-step sequence of aldol cyclopropanation-retro-aldol reactions for the stereoselective synthesis of enantiopure cyclopropane carboxaldehydes. PMID- 15877133 TI - Nanometer-scale ordering in cast films of columnar metallomesogen as revealed by STM observations. AB - STM observations were performed on a cast film of a columnar metallomesogen ([Cr(5C8)3]; 5C8 = 1-(3,4,5-trioctyloxyphenyl)-3-(3,4-dioctyloxyphenyl)propane 1,3-dionate anion) on a graphite surface, revealing the nanometer-scale surface ordering into an oblique lattice (a = 10.5 nm, b = 11.5 nm, alpha = 55 degrees) possibly due to the DeltaLambda-chiral interactions. PMID- 15877134 TI - Semiconducting polyfluorenes with electrophosphorescent on-chain platinum-salen chromophores. AB - The synthesis of statistical fluorene-type copolymers with on-chain Pt-salen phosphorescent units and their use in electrophosphorescent OLEDs is reported. PMID- 15877135 TI - Formation of GeO2 nanosheets using water thin layers in lamellar phase as a confined reaction field--in situ measurement of SAXS by synchrotron radiation. AB - Highly crystallized GeO2 nanosheets were synthesized by hydrolysis and condensation reactions of germanium alkoxide using a 2-dimensional flat thin lamellar phase water layer containing surfactant molecules at the liquid-liquid interface as a confined reaction field. PMID- 15877136 TI - Kinetics of polycondensation reactions during self-assembly of mesostructured films studied by in situ infrared spectroscopy. AB - In situ synchrotron FTIR experiments have been performed during evaporation induced self-assembly (EISA) of mesoporous films and the role of silica polycondensation in obtaining highly organized mesostructures has been illuminated. PMID- 15877137 TI - Polygonal gold nanoplates in a polymer matrix. AB - Polygonal gold nanoplates are generated in situ in poly(vinyl alcohol) film through thermal treatment, the polymer serving as the reducing agent and stabilizer for the nanoparticle formation and enforcing preferential orientation of the plates. The rare pentagonal as well as the more commonly observed hexagonal, triangular and square/rectangle shapes are obtained by fine-tuning the Au/PVA ratio and the time and temperature of fabrication. PMID- 15877138 TI - Dendrimeric Gd(III) complex of a monophosphinated DOTA analogue: optimizing relaxivity by reducing internal motion. AB - A marked increase of relaxivity has been observed upon rigidifying the internal frame of Gd-containing PAMAM dendrimers: the effect has been attained by either protonation of the dendrimer or by forming supramolecular adducts with cationic polyaminoacids. PMID- 15877139 TI - Palladium catalysed enantioselective phosphination reactions using secondary phosphine-boranes and aryl iodide. AB - Preliminary results dealing with the enantioselective version of the C-P cross coupling reaction between dissymmetric secondary phosphine-borane complexes and aryl iodide derivatives are presented. To gain information on the enantiodiscriminating step, direct observation of an intermediate involved in the catalytic cycle has been achieved by (31)P NMR spectroscopy. PMID- 15877140 TI - New families of supermicroporous metal oxides: the link between zeolites and mesoporous materials. AB - Sol-gel hydrolysis reactions in propanol of two or more metal acetates or alkoxides in n-alkylamines have been found to yield porous mixed oxides with the presence of pores largely in the 10-20 A region. PMID- 15877141 TI - One-pot beta-substitution of enones with alkyl groups to beta-alkyl enones. AB - Vinylic hydrogens at the beta-position of enones were effectively substituted with alkyl groups in a one-pot procedure to afford beta-alkyl enones in good to high isolated yields by conjugate addition of higher-order dialkyl cyanocuprates to enones, followed by a reaction with N-tert-butylbenzenesulfinimidoyl chloride at -78 degree C. PMID- 15877142 TI - A three-dimensional porous metal-organic framework with the rutile topology constructed from triangular and distorted octahedral building blocks. AB - The solvothermal reaction of zinc acetate dihydrate with 1,3,5 benzenetricarboxylic acid yields a three-dimensional porous metal-organic framework constructed from triangular and distorted octahedral building blocks, the framework of which can be described as a decorated rutile net. PMID- 15877143 TI - Influence of hydrogen bonding on coordination polymer assembly. AB - An extended dipyridyl ligand (L1) capable of hydrogen bonding with guest species via urea functionalities has been designed and synthesised. Assembly of a silver(I) coordination polymer of L1 is dependent on the nature of the hydrogen bond acceptor in a logical extension of the monopyridyl analogue. PMID- 15877144 TI - A comparison of the solid state structures of the selenium(IV) compounds PhSeX3(X=Cl, Br). AB - The crystal structures of PhSeX3(X=Cl, Br,) and their spectroscopic data are reported, with the structure of PhSeBr3 exhibiting interesting molecular, charge transfer, and ionic bonding aspects. PMID- 15877145 TI - New stoichiometries from an old system: amine complexes of silver chloride. AB - The unexpected stoichiometries L4(AgCl)5 and L4(AgCl)3 have been established for complexes with simple amines L. PMID- 15877146 TI - Selection of the cis and trans phosph(III)azane macrocycles [{P(mu-NtBu)}2(1-Y-2 NH-C6H4)]2(Y=O, S). AB - The 1 : 1 reactions of [ClP(mu-NtBu)]2 with the difunctional aromatic amines 1,2 1-YH-2-NH2-C6H4 in the presence of Et3N give the dimeric phosph(III)azane macrocycles [{P(mu-NtBu)2(1-Y-2-HN-C6H4)]2, predominantly as the cis isomer in the case of Y=O (1.cis) and as the trans isomer for Y=S (2.trans). Model M.O. calculations suggest that the selection of the cis and trans isomers is not thermodynamically controlled. The alternative isomers 1.trans and 2.cis are generated exclusively by the deprotonation of the model intermediates [(1-Y-2-NH2 C6H4)P(mu-NtBu)]2[Y=O (3), S (4)] with nBuLi followed by cyclisation with [ClP(mu NtBu)]2. The solid-state structures of 1.cis/trans(50 : 50), 2.cis, 3 and 4 are reported. PMID- 15877147 TI - Structural variation in organically templated uranium sulfate fluorides. AB - The ability of templated uranium sulfate fluorides to adopt diverse inorganic architectures is demonstrated in six novel materials. The inorganic structures present in [N2C6H16][UO2F2(SO4)](USFO-2), [N2C6H16][UO2F(SO4)]2(USFO-3), [N2C3H12][UO2F(SO4)]2.H2O (USFO-4), [N2C5H14][UO2F(H2O)(SO4]2(USFO-5), [N2C6H18]2[UO2F(SO4)]4.H2O (USFO-6) and [N2C3H12][UO2F(SO4)]2.H2O (USFO-7) range from infinite chains to five different layer topologies. The chain, and two of the five layers, have unprecedented structure types. These compounds illustrate the structural diversity within this new family of materials, arising from the varied coordination of the U6+ centres. Each material was synthesised under hydrothermal conditions, through reaction of uranyl acetate, sulfuric acid, HF(aq), water, and the respective organic template. PMID- 15877148 TI - Reductions by aquatitanium(II). AB - Solutions of titanium(II), prepared by dissolving titanium wire in mixtures of hydrofluoric and triflic acids, reduce quinones, nitrosodisulfonate anion, and complexes of cobalt(III). When the oxidant is taken in excess, these reactions yield Ti(IV), whereas with excess reductant, the principal product is Ti(III). These reactions are compared with those by Ti(III). Despite differences in rate laws, it is clear that rate ratios for the two reductants (kTiII/kTiIII) fall well below 10(4), the minimum selectivity corresponding to estimated differences in formal potentials, and in some instances, Ti(II), the stronger reductant, reacts more slowly. For both Ti(III) and Ti(II), reductions within the series [Co(NH3)5X]2+(where X=F, Cl, Br, and I), the fluoro complex reacts much more rapidly than its congeners, and the bromo and iodo complexes are slowest, an order similar to that for Eu2+ reductions, but opposite to that for Cr(II) and Cu(I). The [Co(NH3)5Br]2+ reaction with excess Ti(II) proceeds at rates very nearly independent of [oxidant] during the first 80-90% reaction, implying that initiation occurs via unimolecular conversion of Ti(II) to an activated cationic reducing species, in the same manner as the earlier described reduction of I3- by Ge(II) in aqueous HCl. PMID- 15877149 TI - Synthesis, reactivity and structural studies of carboranyl thioethers and disulfides. AB - The equimolar reaction of 1-SH-2-R-1,2-closo-C2B10H10(R=Me, H, Ph) with KOH in ethanol produces the thiolate species [1-S-2-R-1,2-closo-C2B10H10]-. These react with iodine to give the disulfide bridged dicluster (1-S-2-R-1,2-closo C2B10H10)2(R=H, Me, Ph) compounds as analytically pure, white and air-stable solids in high yield. Synthesis of monothioether bridged species is synthetically more difficult. In fact three procedures have been tested to obtain the thioether bridged dicluster compounds (2-R-1,2-closo-C2B10H10)2S (R=Me, H, Ph) but only (2 Me-1,2-closo-C2B10H10)2S was successfully synthesized and characterized. Attempts to produce mixed compounds (1-R-1,2-closo-C2B10H10)S(1-R'-1,2-closo-C2B10H10), R not=R', were unsuccessful. Deboronation reaction of this dicarboranylthioether lead, depending on the reaction conditions, to monoanionic [(2-Me-1,2-closo C2B10H10)S(8-Me-7,8-nido-C2B9H10)]- or dianionic [(8-Me-7,8-nido-C2B9H10)2S]2- sulfur bridge anions. Deboronation of carboranyl disulfides gave the corresponding dianionic [(7-S-8-R-7,8-nido-C2B9H10)2]2-(R=H, Me, Ph) species. This reaction was very dependent, however, on the reaction conditions. With slight variation of the reaction conditions, splitting of the S-S bond leading to the thiolate species with retention of the closo cluster was also found. Carboranyl disulfides (1-S-2-R-1,2-closo-C2B10H10)2(R=H, Me, Ph) do not lead to thiosulfinates R-S(O)-S-R' by oxidation with H2O2 or I2 as organic disulfides do. This behaviour is attributed to the presence of the sulfur atom directly bonded to the carbon cluster that produces electronic transfer from the filled orbitals on the sulfur atom into the cage LUMO (largely located on the cage Cc-Cc bond). This causes a depletion of electron density on the sulfur, thence impairing sulfur oxidation, and facilitating S-S breaking. Crystal structures of monothioethers (2-Me-1,2-closo-C2B10H10)2S, [NMe4][(2-Me-1,2-closo-C2B10H10)S(8 Me-7,8-nido-C2B9H10)](the first example reported in the literature of a two cluster compound incorporating the closo C2B10 and the nido[C2B9]- moieties linked by a one member spacer) and disulfides (1-S-1,2-closo-C2B10H11)2, (1-S-2 Me-1,2-closo-C2B10H10)2, (1-S-2-Ph-1,2-closo-C2B10H10)2 are reported which support the behaviour of these species. PMID- 15877150 TI - Preparation of hydrido(vinylidene)ruthenium(II) complexes and a one-pot synthesis of Grubbs-type ruthenium carbenes. AB - Treatment of the hydrido(dihydrogen) compound [RuHCl(H2)(PCy3)2] 1 with alkynes RC[triple bond, length as m-dash]CH (R=H, Ph) afforded the hydrido(vinylidene) complexes [RuHCl(=C=CHR)(PCy3)2] 2, 3 which react with HCl or [HPCy3]Cl to give the corresponding Grubbs-type ruthenium carbenes [RuCl2(=CHCH2R)(PCy3)2] 4, 5. The reaction of 2 (R=H) with DCl, or D2O in the presence of chloride sources, led to the formation of [RuCl2(=CHCH2D)(PCy3)2] 4-d1. Based on these observations, a one-pot synthesis of compounds 4 and 5 was developed using RuCl3.3H2O as the starting material. The hydrido(vinylidene) derivative 2 reacted with CF3CO2H and HCN at low temperatures to yield the carbene complexes [RuCl(X)(=CHCH3)(PCy3)2] 6, 7, of which 7 (X=CN) was characterized crystallographically. Salt metathesis of 2 with CF3CO2K and KI led to the formation of [RuH(X)(=C=CH2)(PCy3)2] 8, 9. The bis(trifluoracetato) and the diiodo compounds [RuX2(=CHCH3)(PCy3)2] 10, 11 as well as the new phosphine P(thp)3 12 (thp=4-tetrahydropyranyl) and the corresponding complex [RuCl2(=CHCH3){P(thp)3}2] 14 were also prepared. The catalytic activity of the ruthenium carbenes 4-7, 10, 11 and 14 in the olefin cross-metathesis of cyclopentene and allyl alcohol was investigated. PMID- 15877151 TI - The synthesis, structure and properties of copper(II) complexes of asymmetrically functionalized derivatives of 1,4,7-triazacyclononane. AB - Reaction of 1-propylamino-4-acetato-1,4,7-triazacyclononane (L1), 1-benzyl-4 acetato-1,4,7-triazacyclononane (L2) and 1-benzyl-4-propylamino-1,4,7 triazacyclononane (L3) with a copper(II) salt gave Na2[CuL1](ClO4)3(1a), [CuL2]Cl (2) and [Cu2L32](ClO4)4.5H2O (3), respectively. [CuL4]ClO4 (4) was formed by reacting 1-formyl-4-ethylacetato-1,4,7-triazacyclononane with cupric chloride in aqueous solution. The X-ray crystal structures of the complexes reveal that the ligands generate distorted square pyramidal or square planar coordination environments about the Cu(II) centre, but in three complexes (1b, 3 and 4) weak interactions to an oxygen atom from a perchlorate anion and, in the case of 4, also to an amide nitrogen leading to tetragonally elongated octahedral Cu(II) geometries. In 4, the formyl group is found to reduce the coordinating ability of the macrocyclic nitrogen to which it is attached, as evidenced by the weak CuN interaction. The formation of five-membered chelate rings on coordination of the ligands further contributes to the distortion from the ideal geometries. The crystal lattices contain a number of novel supramolecular features. 1a contains a negatively charged sodium perchlorate chain of composition [Na2(ClO4)3]x(x-), with a complex series of Na-O-Na bridges flanked by [CuL1]+ units, while 3 contains highly complex hydrogen bonded sheets approximately 20 A thick that stack through van der Waals interactions. One-dimensional chains comprised of copper complexes are found in 2 and 4, and are held together by hydrogen bonds in 2 and acetate bridges between the copper cations in 4. The solution EPR spectra indicate that the copper(II) centres exist in isolated distorted square pyramidal (possibly square planar for 4) environments, while in the solid state there is evidence for the existence of weak exchange and dipole-dipole coupling for some complexes. PMID- 15877152 TI - The mineral phase in the cuticles of two species of Crustacea consists of magnesium calcite, amorphous calcium carbonate, and amorphous calcium phosphate. AB - The cuticules (shells) of the woodlice Porcellio scaber and Armadillidium vulgare were analysed with respect to their content of inorganic material. It was found that the cuticles consist of crystalline magnesium calcite, amorphous calcium carbonate (ACC), and amorphous calcium phosphate (ACP), besides small amounts of water and an organic matrix. It is concluded that the cuticle, which constitutes a mineralized protective organ, is chemically adapted to the biological requirements by this combination of different materials. PMID- 15877153 TI - Synthesis, crystal structure and photoconductivity of the first [60]fullerene complex with metal diethyldithiocarbamate: {CuII(dedtc)2}2.C60. AB - The first molecular complex of fullerene C60 with metal dithiocarbamate, namely, {CuII(dedtc)2}2.C60(dedtc: diethyldithiocarbamate) (1) was obtained as single crystals. Butterfly-shaped CuII(dedtc)2 molecules efficiently co-crystallized with spherical fullerene molecules to form a layered structure, in which closely packed hexagonal C60 layers alternate with the layers composed of CuII(dedtc)2 dimers. The formation of the complex with C60 changes geometry and the EPR spectrum of starting CuII(dedtc)2. Magnetic susceptibility of 1 follows the Curie Weiss law in the 300-1.9 K range with the negative Weiss constant of -2.5 K showing a weak antiferromagnetic interaction between CuII centers in the dimers. The crystals of 1 have low dark conductivity of 10(-11) S cm-1, which is consistent with a neutral ground state of the complex. Illumination of the crystals by white light increases the photocurrent by 20-50 times. The photoconductivity spectrum of 1 has a maximum at 470 nm showing that both intermolecular charge transfer between neighboring C60 molecules and photoexcitation of CuII(dedtc)2 can contribute to photogeneration of free charge carriers. The effect of a weak magnetic field with Bo<0.5 T on the photoconductivity of 1 has been found. PMID- 15877154 TI - The methoxycarbonylation of aryl chlorides catalysed by palladium complexes of bis(di-tert-butylphosphinomethyl)benzene. AB - A catalyst system based on palladium-1,2-bis-(di-tert butylphosphinomethyl)benzene (BDTBPMB) shows good activity for the methoxycarbonylation of strongly activated aryl chlorides, like 4 chloromethylbenzoate or 4-chlorocyanobenzene. Surprisingly, the use of less activated aryl chlorides, like 4-chloroacetophenone, leads to the formation of dimethyl terephthalate amongst other products arising from organic reactions of methoxide ion and/or CO. Less nucleophilic alcohols such as 2,2,2 trifluoroethanol promote the formation of carbonylation products even from 4 chloroacetophenone and chlorobenzene. Labelling studies involving CD3OH, CD3OD or 13CO give information on the origin of many of the products. PMID- 15877155 TI - Investigation of the reactivity of arylamines, organo-hydrazines and tolylisocyanate towards [PW12-xMxO40]n- Keggin anions. AB - Reaction of K7[A,alpha-PW9Mo2O39] with Na2MoO4.2H2O in a mixture of water/dioxane/hydrochloric acid and further precipitation with (Bu4N)Br provided (Bu4N)3[A,alpha-PW9Mo3O40](3). Analogous reaction with K7-xNax[alpha-PW11O39] is an alternative to the synthesis of (Bu4N)3[alpha-PW11O39{MoVIO}]2. Multinuclear NMR and ESI mass spectrometry have been used to interpret the reaction of (Bu4N)x[alpha-PW11O39{ReO}](x=3 1; x=4 1I), (Bu4N)x[alpha-PW11O39{MoO}](x=3 2; x=4 2I) and (Bu4N)3[A,alpha-PW9Mo3O40]3 by organohydrazines, arylamines, tolylisocyanate and tetraphenylphosphine imide. PMID- 15877156 TI - The synthesis and characterisation of 4,1,2-MC2B10 metallacarboranes. AB - Reduction of the tethered carborane 1,2-(CH2)3-1,2-closo-C2B10H10 followed by treatment with CoCl2/NaCp, [(p-cymene)RuCl2]2(p-cymene=C6H4MeiPr-1,4), (PMe2Ph)2PtCl2 or (dppe)NiCl2(dppe=Ph2PCH2CH2PPh2) affords reasonable yields of the new 13-vertex metallacarboranes 1,2-(CH2)3-4-Cp-4,1,2-closo-CoC2B10H10 (1), 1,2-(CH2)3-4-(p-cymene)-4,1,2-closo-RuC2B10H10 (2), 1,2-(CH2)3-4,4-(PMe2Ph)2 4,1,2-closo-PtC2B10H10 (3) and 1,2-(CH2)3-4,4-(dppe)-4,1,2-closo-NiC2B10H10 (4), respectively. All compounds were characterised spectroscopically and crystallographically. The cobalt and ruthenium species 1 and 2 have Cs symmetry in both solution and the solid state, having henicosahedral cage structures featuring a trapezoidal C1C2B9B5 face. The platinum and nickel compounds 3 and 4 have asymmetric docosahedral cage structures in the crystal (the more so for 4 than for 3) although both appear, by 11B and 31P NMR spectroscopy, to have Cs symmetry in solution. Low-temperature experiments on the more soluble platinacarborane could not freeze out the diamond-trapezium-diamond fluctional process that we assume is operating in solution, and we therefore conclude that this process has a relatively low activation barrier, probably <35 kJ mol-1. PMID- 15877157 TI - Addition products of a P(III)-isothiocyanate to dialkyl acetylenedicarboxylates: a spirocyclic phosphinimine and a triphosphorus heterocycle with tetra- and penta coordinate phosphorus. AB - Treatment of the P(III) isothiocyanate CH2[6-t-Bu-4-Me-C6H2O]2PNCS (1) with dimethyl acetylenedicarboxylate (DMAD) or diethyl acetylenedicarboxylate (DEAD) yields the spirocyclic phosphinimines CH2[6-t-Bu-4-Me C6H2O]2P[NC(S)C(CO2R)C(CO2R)][R=Me (2), Et (3)], in a reaction unlike those of organic isocyanates. From the reaction of 1 with DEAD, a second product, the triphosphorus compound 5, with the composition [2x1+3] but with a completely reorganized structure {CH2[6-t-Bu-4-Me-C6H2O]2P=C(CO2Et)C(CO2Et)=CN-}{CH2[6-t-Bu 4-Me-C6H2O]2P(NCS)}-SC=N-P(S)[(OC6H2-6-t-Bu-4-Me)2CH2] with tetra- and penta coordinate phosphorus, is also isolated. Structure and reactivity of these compounds are discussed. Addition of 2,2,2-trifluoroethanol to 2 or 3 leads to the pentacoordinate phosphorus compounds [CH2(6-t-Bu-4-Me C6H2O)2P(OCH2CF3){C(CO2R)C(CO2R)-C(S)-NH-}][R=Me (6), Et (7)]. The phosphonate [CH2(6-t-Bu-4-Me-C6H2O)2P(O)C(CO2Et)=C(CO2Et)-C(S)-NH2] (8) is obtained by evaporating a solution of 7 in open air. PMID- 15877158 TI - Studies of oxalate-bridged MM quadruple bonds and their radical cations (M=Mo or W): on the matter of linkage isomers. AB - Electronic structure calculations employing density functional theory (DFT) and time-dependent density functional theory (TD-DFT) have been carried out on the model complexes {[(HCO2)3M2]2(mu-O2CCO2)}0/+(M=Mo or W) in D2h symmetry, where the oxalate bridge forms either five- or six-membered rings with the M(2) centres; the complexes are hereafter referred to as mu(5,5)0/+ and mu(6,6)0/+, respectively. The calculations predict that the neutral complexes should exist as the mu(5,5) linkage isomer, while the radical cations favour the mu(6,6) isomer by ca. 4-6 kJ mol-1. For the mu(5,5) isomers, the rotational barriers about the oxalate C-C bond have been calculated to be 15.9 and 27.2 kJ mol-1 for M=Mo and W, respectively. For the cationic mu(5,5)+ isomers the barrier is higher, being 36.8 and 50.6 kJ mol-1 for M=Mo and W, respectively. The calculated Raman and visible near-IR spectra for the mu(5,5)0/+ and mu(6,6)0/+ are compared with experimental data obtained for the {[(tBuCO2)3M2]2(mu-O2CCO2)}0/+ complexes, hereafter referred to as M4OXA0/+(M=Mo or W). The experimental data more closely correlate with that calculated for the mu(5,5)0/+ linkage isomers, and the 13C NMR spectrum of the mixed metal complex Mo2W2OXA indicates the presence of the 5 membered oxalate-bridged species (J(CC)=100 Hz). PMID- 15877159 TI - Density functional theory study of eight-atom germanium clusters: effect of electron count on cluster geometry. AB - Density functional theory (DFT) at the hybrid B3LYP level has been applied to the germanium clusters Ge8z(z=-6, -4, -2, 0, +2, +4) using nine initial geometries. For Ge8(2-) the D2d bisdisphenoid structure predicted by the Wade-Mingos rules is not computed to be the global minimum but instead lies 3.9 kcal mol-1 above the Td tetracapped tetrahedron global minimum predicted to exhibit spherical aromaticity. The hyperelectronic clusters Ge(8)4- and Ge8(6-) have nido B8H12 and square antiprism structures, respectively, as global minima in accord with the Wade-Mingos rules and experimental data on E(8)2+(E=Sb, Bi) cations. Hypoelectronic eight-vertex clusters isoelectronic and isolobal with Ge8, Ge8(2+) and Ge(8)4+ are not known experimentally. Their computed structures include smaller polyhedra having one or more capped triangular faces as well as more open non-polyhedral structures. PMID- 15877160 TI - The analysis of flue gas treatment residues using non-destructive X-ray fluorescence as a regulatory compliance test. AB - Current and proposed European Union (EU) regulations require the residual material from municipal solid waste incineration to be characterised prior to disposal. X-Ray fluorescence (XRF) provides a rapid and non-destructive technique for analysing such materials. PMID- 15877161 TI - Characterization of individual aerosol particles in workroom air of aluminium smelter potrooms. AB - Aerosol particles with aerodynamic diameters between 0.18 and 10 microm were collected in the workroom air of two aluminium smelter potrooms with different production processes (Soderberg and Prebake processes). Size, morphology and chemical composition of more than 2000 individual particles were determined by high resolution scanning electron microscopy and energy-dispersive X-ray microanalysis. Based on chemical composition and morphology, particles were classified into different groups. Particle groups with a relative abundance above 1%(by number) include aluminium oxides, cryolite, aluminium oxides-cryolite mixtures, soot, silicates and sea salt. In both production halls, mixtures of aluminium oxides and cryolite are the dominant particle group. Many particles have fluoride-containing surface coatings or show agglomerations of nanometer sized fluoride-containing particles on their surface. The phase composition of approximately 100 particles was studied by transmission electron microscopy. According to selected area electron diffraction, sodium beta-alumina (NaAl(11)O(17)) is the dominant aluminium oxide and cryolite (Na(3)AlF(6)) the only sodium aluminium fluoride present. Implications of our findings for assessment of adverse health effects are discussed. PMID- 15877162 TI - Theoretical aspects of fluoride air contaminant formation in aluminium smelter potrooms. AB - The amount of particulate fluorides evolved from aluminium electrolysis cells is not entirely accounted for by the fluorides entrained in the anode gas. The largest additional source of particulate fluoride formation is by direct evaporation of fluorides into the anode gas stream and subsequent condensation on the drops of electrolyte generated in the process of bubble burst. A theoretical model was used for the calculation of the main physical parameters responsible for the formation of particle nuclei when the hot anode-gas is mixed with ambient air. The results of these calculations are in agreement with experimental observations reported in the literature. In particular, the size distribution, composition and morphology of the nano-particles support the theory of a vapour condensation mechanism under conditions of extreme supersaturation, but further studies are necessary. PMID- 15877163 TI - Investigation of the concentration and isotopic composition of inputs and outputs of Pb in waters at an upland catchment in NE Scotland. AB - As a consequence of the accumulation of anthropogenic Pb in upland catchments, there has been much recent concern about the potential mobilisation and transport of Pb from the soils to receiving waters and also the possible harmful effects that this might have on aquatic biota. This paper presents the findings of a two year study of Pb behaviour in an organic-rich upland catchment at Glensaugh in NE Scotland. Pb inputs to the catchment were characterised by direct measurements of Pb concentration and (206)Pb/(207)Pb ratios in rain water and interception. Pb outputs from the catchment were calculated from measurements on stream water samples taken from the two main streams, the Cairn Burn and Birnie Burn. The relative contribution of Pb from groundwater and throughflow, under different flow conditions (base flow and high flow), to stream waters was investigated via analysis of springs sourced from groundwater and of waters flowing through the various soil horizons (S (surface), A, B, C, and D), respectively. The outcome of intensive sampling and analysis over the two-year time period was that, even with marked reduction in Pb inputs over the past two decades, the catchment was still acting as a net sink for the current atmospheric deposition. Although the Pb isotopic signature for stream water is very similar to that for the contemporaneous rain water ((206)Pb/(207)Pb approximately 1.15-1.16), only a small portion of the rain water is transferred directly to stream water. Instead, the Pb input is transferred to the stream waters mainly via groundwater and it was also confirmed that the latter had a similar Pb isotopic signature. From the Pb isotopic measurements on throughflow waters, however, Pb being removed via the streams contained some previously deposited Pb, i.e. mobilisation of a small portion of soil-derived anthropogenic Pb was occurring. These findings are important not only with respect to the source/sink status of the catchment but also for calculation of the extent of retention of the current atmospheric Pb inputs, which must take account of the release of previously deposited Pb from the catchment soils, a process occurring mainly under high flow conditions. PMID- 15877164 TI - The estimation of physicochemical properties of methyl and other alkyl naphthalenes. AB - The GLC and HPLC data of Autenrieth and co-workers (P. Dimitriou-Christidis, B. C. Harris, T. J. McDonald, E. Reese and R. L. Autenrieth, Chemosphere, 2003, 52, 869) has been used to obtain solvation descriptors for methyl naphthalenes for use in the Abraham solvation equations. These descriptors are then used to predict a large number of physicochemical properties, of environmental importance. These include solubility in water and the gas-water partition coefficient (equivalent to the Henry's Law constant for the water to gas partition). Predictions are in excellent agreement with those of Autenrieth and co-workers and with experimental observations, where available. Other important predictions are the gas-dry octanol and gas-wet octanol partition coefficients. Descriptors have also been obtained for the ethyl-, propyl- and butyl naphthalenes which, again, can be used to predict numerous physicochemical properties. PMID- 15877165 TI - Large particles are responsible for elevated bacterial marker levels in school air upon occupation. AB - Muramic acid (Mur) is found in bacterial peptidoglycan (PG) whereas 3-hydroxy fatty acids (3-OH FAs) are found in Gram-negative bacterial lipopolysaccharide (LPS). Thus Mur and 3-OH FAs serve as markers to assess bacterial levels in indoor air. An initial survey, in a school, demonstrated that the levels of dust, PG and LPS (pmol m(-3)) were each much higher in occupied rooms than in the same rooms when unoccupied. In each instance, the Mur content of dust was increased and the hydroxy fatty acid distribution changed similarly suggesting an alteration in the bacterial population. Here, findings are compared with results from two additional schools. Follow-up aerosol monitoring by particle size was also performed for the first time for all 3 schools. The particle size distribution was shown to be quite different in occupied versus unoccupied schoolrooms. Within individual classrooms, concentrations of airborne particles [greater-than-or-equal]0.8 [micro sign]m in diameter, and CO(2) were correlated. This suggests that the increased levels of larger particles are responsible for elevation of bacterial markers during occupation. Release of culturable and non culturable bacteria or bacterial aggregates from children (e.g. from flaking skin) might explain this phenomenon. PMID- 15877166 TI - In vivo K-shell X-ray fluorescence bone lead measurements in young adults. AB - The (109)Cd K-shell X-ray fluorescence (XRF) technique was used to measure in vivo tibia lead concentrations of 34 young adults living in the state of Vermont (USA) and the province of New Brunswick (Canada). The subjects ranged in age from 18 to 35 years, and had no known history of elevated lead exposure. Measurement parameters were varied, using the same XRF system for both populations. Tibia lead concentrations were low for both groups, with mean values of 0.7 microg lead g(-1) bone mineral (Vermont) and 0.5 microg g(-1)(New Brunswick). No individual measurement exceeded 7 microg g(-1). Mean uncertainty values obtained for the Vermont and New Brunswick subjects were 4.1 microg g(-1) and 2.6 microg g(-1), respectively. Improved measurement uncertainty in the New Brunswick group was attributed to the use of a reduced source-to-skin distance (approximately 5 mm) and a longer measurement time (3600 seconds) using a weaker radioisotope source (< or =0.42 GBq). Measurement uncertainty tended to increase with body mass index. For a given body mass index, female subjects returned a measurement uncertainty approximately 1 microg g(-1) greater than males. PMID- 15877167 TI - Assessment of workers' exposure to palladium in a catalyst production plant. AB - Airborne particulate matter was collected and biomonitoring of workers was performed by sampling blood, urine and hair of 84 exposed subjects, 17 occasionally exposed employees, 21 controls from administrative offices and 25 unexposed people (external controls). Determination of Pd was performed using Quadrupole and High Resolution Inductively Coupled Plasma Mass Spectrometry. The Production of Catalysts Department and the Refining Service presented the highest levels of Pd in airborne matter collected by means of an area sampler. The highest level of soluble Pd (1.66 microg m(-3)) was found in the Production of Catalysts Department. The highest concentration of Pd in airborne matter, collected by means of personal devices (7.90 microg m(-3)) was found in the Refining Service. Hair showed a clear distribution pattern among departments, with values ranging from 0.60 to 5.54 microg g(-1). Administrative workers presented blood levels of Pd between 2 and 500 times higher than external controls. Only urine levels correlated with the measurements of airborne Pd collected with personal devices. A very strong association between airborne Pd collected by personal devices and Pd levels in hair (r(2)= 0.569, with p< or = 0.01) and urine (r(2)= 0.684, with p< or = 0.01) was found. On the basis of these findings: (i) blood results appear to be an unsuitable biological marker for occupational exposure to Pd; (ii) urine could be considered as a satisfactorily responsive bio-marker for occupational monitoring; and (iii) hair cannot be considered a good index of time-related exposure. PMID- 15877168 TI - A chemosorptive cylindrical denuder designed for personal exposure measurements of isocyanates-evaluation on generated aerosols of 4,4'-methylenediphenyl diisocyanate. AB - A denuder/filter system constructed for solvent-free personal exposure measurements was evaluated for separation of vapour and particulate 4,4' methylenediphenyl diisocyanate (4,4'-MDI) generated from heated PUR-foam. The two different phases were collected in the denuder and on the filter, respectively, by chemosorption on a polydimethylsiloxane (SE-30)-dibutylamine (DBA) stationary phase. Both repeatability and the total mass concentration of 4,4'-MDI were similar to that obtained from the reference method, in this case an impinger/filter system. The penetration of particles through the denuder at 300 ml min(-1) was nearly 100% in the particle size range 25 to 700 nm, which fits well with the Gormley-Kennedy equation. Denuder/filter sampling of the 4,4'-MDI aerosol at 500 ml min(-1) yielded a phase distribution that was in accordance with the results from the reference method. The method limit of detection was 6 ng m(-3) and 4 ng m(-3) for the denuder and filter, respectively, when using an air sampling flow rate of 300 ml min(-1) and a sampling period of 15 min. This is well below the Swedish occupational exposure limit (OEL) of 50 and 100 microg m( 3) for an 8-hour working day and a 5-min period, respectively. PMID- 15877169 TI - The effect of filter material on bioaerosol collection of Bacillus subtilis spores used as a Bacillus anthracis simulant. AB - The objective of this study was to determine filter materials and extraction methods that are appropriate to use for environmental sampling of B. anthracis. Four types of filters were tested: mixed cellulose ester (MCE) with a pore size of 3 microm, polytetrafluoroethylene (PTFE) with pore sizes of 1 and 3 microm, and gelatin with a pore size of 3 microm. Bacillus subtilis var. niger endospores (also known as Bacillus globigii[BG]) were used as a surrogate for B. anthracis. Endospores were collected into Button Inhalable Aerosol Samplers with sampling times of 15 minutes, 1 hour, and 4 hours. Physical collection efficiency was determined by measuring upstream and downstream B. subtilis concentrations with an optical particle counter. Vortexing with ultrasonic agitation and vortexing with shaker agitation extraction methods were evaluated. The MCE, 1 microm PTFE, and gelatin filters provided physical collection efficiencies of 94% or greater. The 3 microm PTFE filter showed inconsistent physical efficiency characteristics between filters. Epifluorescence microscopic analysis of the gelatin filter extraction fluid revealed the presence of contamination by non-culturable bacteria. Mean differences for microbial culturability were not statistically significant for filter materials and extraction methods. However, the vortexing with shaker agitation extraction method resulted in higher total microbial counts in the extraction fluids for MCE and 1 microm PTFE filters when compared to vortexing with ultrasonic agitation. In summary, the MCE and 1 microm PTFE filters in combination with vortexing and shaker extraction demonstrated the best performance for the filter collection and extraction of BG spores. PMID- 15877170 TI - Aspiration and sampling efficiencies of the TSP and louvered particulate matter inlets. AB - An experimental system was developed for the rapid measurement of the aspiration/transfer efficiency of aerosol samplers in a wind tunnel. We attempted to measure the aspiration and particle transfer characteristics of two inlets commonly used for sampling airborne Particulate Matter (PM): the 'Total Suspended Particulate' or TSP inlet, and the louvered 'dichotomous sampler inlet' typically used in sampling PM10 or PM2.5. We were able to determine the fraction of the external aerosol that enters the inlet and is transferred through it, and hence is available for collection by a filter, or further size fractionation into PM10 or PM2.5. This 'sampling efficiency' was analysed as a function of dimensionless aerodynamic parameters in order to understand the factors governing inlet performance. We found that for the louvered inlet the sampling efficiency increases as the external wind increases. Under all conditions expected in practical use the louvered inlet aspirates sufficient PM to allow either PM10 or PM2.5 to be selected downstream. The TSP inlet's sampling efficiency decreases with increasing external wind, and the TSP inlet is likely to under-sample the coarse end of the PM10 fraction at moderate and high external winds. As this inlet is generally not used with a downstream size fractionator, changes in sampling efficiency directly affect the measured aerosol concentration. We also investigated whether it is possible to dimensionally scale the PM inlets to operate at either higher or lower flow rates, while preserving the same sampling characteristics as the current full-scale, 16.67 L min(-1) versions. In the case of the louvered inlet, our results indicate that scaling to lower flow rates is possible; scaling to higher flow rates was not tested. For the TSP sampler, the sampling efficiency changes if the sampler is scaled to operate at smaller or larger flow rates, leading to unreliable performance. PMID- 15877171 TI - Atmospheric methane at Zeppelin Station in Ny-Alesund: presentation and analysis of in situ measurements. AB - In situ methane (CH(4)) measurement techniques and data from the Zeppelin Station in Ny-Alesund on Svalbard (N 78 degrees 54' E 11 degrees 53') on Mt. Zeppelin (475 m.a.s.l) are presented. The data span the time period from October 1998 to December 2003, though not continuously. The daily mean was calculated from 96 samples per day which are analysed by gas chromatography with flame ionisation detection. Details of the experimental methods and procedures are given. A harmonic function is fitted to the data with a constant trend and seasonal amplitude. The data are also presented in a regression plot showing the difference between the in situ measurements and flask measurements from the National Oceanic and Atmospheric Administration of the Climate Monitoring and Diagnostics Laboratory (NOAA/CMDL) in Boulder, Colorado (http://www.cmdl.noaa.gov/). The Zeppelin data show typical Arctic air characteristics with wintertime pollution episodes from Europe and Russia and a relatively calm summer state. PMID- 15877172 TI - Measurements of biogenic hydrocarbons and carbonyl compounds emitted by trees from temperate warm Atlantic rainforest, Brazil. AB - This study is part of a three-year project on biogenic volatile organic compound (VOC) emissions from trees of the temperate warm Atlantic rainforest found in the metropolitan area of Sao Paulo City (MASP). No study of VOC emission rates from plant species has been carried out in the temperate warm Atlantic rainforest of Brazil prior to this work. Eleven species were selected (Alchornea sidifolia, Cupania oblongifolia, Cecropia pachystachia, Syagrus romanzoffiana, Casearia sylvestris, Machaerium villosum, Trema micrantha, Croton floribundus, Myrcia rostrata, Solanum erianthum and Ficus insipida) and some of them were studied in urban, sub-urban and forest areas inside the MASP in order to evaluate biogenic VOC composition at sites characterized by different emission sources. Biogenic VOC emissions were determined by placing branches of plants in a dynamic enclosure system, an all-Teflon cuvette, and by sampling the compounds in the air leaving the cuvette. Pre-concentration using adsorbents to retain the VOC, followed by GC-MS after thermal desorption of the sample, was employed to determine the amount of biogenic hydrocarbons. The collection of carbonyl compounds on a 2,4-dinitrophenylhydrazine coated silica followed by HPLC-UV was used to analyze low molecular weight carbonyl compounds. Emission rates of isoprene, alpha-pinene, camphene and limonene ranged from 0.01 to 2.16 microg C h(-1) g(-1) and emission rates of aldehydes (C(2)-C(6)), acrolein, methacrolein and 2-butanone ranged from 1.5 x 10(-2) to 2.3 micro g C h(-1) g (-1). Ambient and leaf temperatures, relative humidity, light intensity, O(3) and NO(x) levels in the local atmosphere were monitored during experiments. It was possible to identify different biogenic VOCs emitted from typical plants of temperate warm Atlantic rainforest. The emission rates were reported as a function of the type of site investigated and were only provided for compounds for which quantification was feasible. Other biogenic compounds were only identified. PMID- 15877173 TI - Performance of semipermeable membrane devices for sampling of organic contaminants in groundwater. AB - Lipid-filled semipermeable membrane devices (SPMDs) are receiving increasing attention as passive, in situ samplers for the assessment of environmental pollutant exposure. Although SPMDs have been successfully used in a variety of field studies in surface waters, only a few studies have addressed their characteristics as groundwater samplers. In this study, the performance of the SPMDs for monitoring organic contaminants in groundwater was evaluated in a pilot field application in an area severely contaminated by chemical waste, especially by chlorinated hydrocarbons. The spatial distribution of hydrophobic groundwater contaminants was assessed using a combination of passive sampling with SPMDs and non-target semiquantitative GC-MS analysis. More than 100 contaminants were identified and semiquantitatively determined in SPMD samples. Along the 6 field sites under investigation, a large concentration gradient was observed, which confirms a very limited mobility of hydrophobic substances in dissolved form in the aquifer. The in situ extraction potential of the SPMD is limited by groundwater flow, when the exchange volume of well water during an exposure is lower than the SPMD clearance volume for the analytes. This study demonstrates that SPMDs present a useful tool for sampling and analyzing of groundwater polluted with complex mixtures of hydrophobic chemicals and provides guidance for further development of passive sampling technology for groundwater. PMID- 15877174 TI - Comparison of TCPP concentrations in sludge and wastewater in a typical German sewage treatment plant-comparison of sewage sludge from 20 plants. AB - Tris(chloro-isopropyl)phosphate (TCPP) was identified by GC-MS by comparing mass spectra and retention times to original standards. The concentrations in wastewater of a sewage treatment plant's influent and effluent were analysed (520 ng l(-1) and 380 ng l(-1), respectively (mean values). The concentrations of TCPP in the wastewater inflow exhibited a high variability. The elimination of this compound in the sewage treatment plant also exhibits a high variability but is low. Additionally the concentrations in sewage sludge of the same plant were determined (mean value 5100 ng g(-1) dry weight; 1700 ng g(-1) wet weight, respectively). For a comparison sludge samples from twenty other plants were analysed. In these samples concentrations ranging from 1000-20000 ng g(-1)(dry weight) were determined. Thus sorption to sludge does occur to some extent. PMID- 15877175 TI - Determination of 226Ra at ultratrace level in mineral water samples by sector field inductively coupled plasma mass spectrometry. AB - An analytical procedure has been proposed for the determination of (226)Ra at the low femtogram per ml concentration level in mineral water samples using double focusing sector field ICP-MS (ICP-SFMS). For the pre-concentration and separation of radium from the matrix elements in water a tandem of a laboratory-prepared filter, based on MnO(2), and Eichrom "Sr-specific" resin was used. The recovery of the method was determined to be 70.5%. The limit of detection for (226)Ra determination was 0.02 fg ml(-1), including a pre-concentration factor of 10. In addition, uranium concentration and uranium isotope ratios were measured by ICP SFMS. In several mineral water samples with a relatively high uranium content, (226)Ra concentrations were found between 0.7-15 fg ml(-1). The effective dose of the contribution was calculated using the radionuclide concentration and dose conversion factors from the World Health Organization, WHO (1993). Assuming a mineral water consumption of 2 l d(-1), a slightly higher calculated dose than the suggested limit for drinking water (0.1 mSv y(-1)) was found in some samples. PMID- 15877176 TI - A method for measuring the potential dermal exposure to methyl methacrylate during two different dental technical work tasks. AB - Dental technicians are exposed on a daily basis to undiluted methyl methacrylate (MMA) when performing various routine tasks. Although the clinical effects of this chemical have been known for decades, no previous studies have been performed to estimate the potential dermal exposure to it. In this study we describe a patch-sampling technique to intercept the MMA that would otherwise have contaminated the skin on different parts of the hand and lower arm. Two different work tasks, making an orthodontic splint and denture preparation, were both performed under simulated workplace conditions twice by two recent graduated dental technicians. Air measurements were collected simultaneously. The results indicate that the exposure patterns associated with producing an orthodontic splint and denture preparation differed. We found work task-dependent differences in the amounts of MMA collected at the different parts of each hand, and differences between the right and left hands. There was also an interaction between hand and work task, especially for the right hand. The air measurements were positively correlated with the dermal exposure. This study highlights the importance of using a measurement strategy that takes the variability within the hand/arm body parts into account when measuring potential exposure during these kinds of work tasks. In order to establish future dermal exposure limits, more workplace and experimental studies are required. PMID- 15877177 TI - Meningioma of the accessory nerve extending from the jugular foramen into the parapharyngeal space. AB - A 35-year-old man presented with pain in the right shoulder and neck for 18 months. The neurological examination revealed complete accessory nerve palsy on the right side without further deficits. Magnetic resonance imaging showed a right parapharyngeal tumour expanding into the posterior fossa through the jugular foramen without dural attachment and absence of invasion into the middle ear cavity or internal auditory meatus. Intraoperative inspection disclosed a tumour originating from the accessory nerve. Histological diagnosis revealed a meningothelial meningioma with invasion of the epineural space. To the knowledge of the authors this is the first report of an accessory nerve meningioma in the jugular foramen associated with a posterior fossa component and extension into the parapharyngeal space. PMID- 15877178 TI - Low-dose methotrexate treatment for oligoarticular juvenile idiopathic arthritis nonresponsive to intra-articular corticosteroids. AB - The objective of this study was to evaluate the efficacy of low-dose (0.2 mg/kg) methotrexate (MTX) in the treatment of children with oligoarticular juvenile idiopathic arthritis (JIA) who do not respond to nonsteroidal anti-inflammatory drugs (NSAIDs) and repeated intra-articular corticosteroid (IA) injections. Nineteen consecutive patients (age: 2-14 years, 18 females) with oligoarticular JIA were studied prospectively. Sixteen had a persistent course and three had an extended course of the disease. Patients were defined as nonresponders to IA injections if the duration of improvement following two consecutive injections was less than 4 weeks. These patients were offered low-dose oral MTX, administered once a week for at least 6 months. Of the 19 patients in this series, 2 responded to NSAIDs alone. Forty-eight IA injections were given to 17 patients; 11 (64%) of them did not respond to this treatment. Nine of the nonresponders were treated with low-dose MTX for a median duration of 15+/-3.8 months. Except for one patient with an extended disease course, all responded very well to treatment and went into remission after a median of 6.4+/-2.9 months, and none required additional IA injections after initiation of MTX treatment. Low-dose oral MTX appears to be very effective in the management of children with oligoarticular JIA, who are unresponsive to IA injections. PMID- 15877179 TI - Clinical and radiological diagnosis of progressive pseudorheumatoid dysplasia in two sisters with severe polyarthropathy. AB - The aim of this case report is to describe unusual cases of progressive pseudorheumatoid dysplasia (PPD) affecting the axial skeleton and peripheral joints and to stress the importance of examining the entire skeleton for definite diagnosis and the importance of rehabilitation interventions. PPD is a rare familial disease characterized by generalized bone-cartilage dysplasia, progressive arthropathy, and platyspondyly. PPD presents as spondyloepiphyseal dysplasia (SED) tarda with progressive arthropathy and progressive pseudorheumatoid arthritis of childhood and is described as a specific autosomal recessive subtype of SED. Two sisters, 18 and 16 years old, with low back pain and polyarthritis are presented. Radiographic and magnetic resonance imaging of the cases revealed typical features characteristic for PPD-like platyspondyly, multiple intravertebral herniations, changes in metaphyses and epiphysis, and mega os trigonum. Consequently, PPD is a rare disease of childhood and should be kept in mind in the differential diagnosis of juvenile idiopathic arthritis to prevent delayed diagnosis and to begin rehabilitation interventions early. It is essential to carefully examine the entire body, particularly the axial skeleton, and to perform radiological evaluation of the spine. These illustrative cases serve to remind physicians to examine the entire skeleton and not to concentrate only on "branches" but also on the "trunk." PMID- 15877180 TI - Idiopathic portal hypertension associated with systemic sclerosis and Sjogren's syndrome. AB - We report a patient with idiopathic portal hypertension (IPH) associated with systemic sclerosis (SSc) and Sjogren's syndrome. A 72-year-old Japanese woman was admitted to our hospital because of Raynaud's phenomenon, sclerodactyly, and dyspnea. The patient had splenomegaly, esophageal varices in the absence of extrahepatic portal obstruction, and cirrhosis of the liver. Immunological studies revealed positive anti-nuclear antibodies and high titers of anti-Scl-70, anti-SS-A, anti-centromere, and anti-mitochondrial M2 antibodies. Histological examinations of the liver biopsy specimen revealed stenosis and loss of small portal veins without findings of primary biliary cirrhosis. The patient was diagnosed as having IPH associated with SSc and Sjogren's syndrome. These observations suggest an immunological role in the pathogenesis of IPH. PMID- 15877181 TI - Dural arteriovenous fistulas presenting with haemorrhagic and ischaemic stroke. PMID- 15877182 TI - Hemifacial spasm due to a subtentorial paramedian meningioma. PMID- 15877183 TI - Italian Right Hemisphere Language Battery: the normative study. AB - Clinical neurolinguistics still lacks consolidated and standardised tools for the assessment of impairments of pragmatics of verbal communication. In the present paper we present norms of the Italian version of the Right Hemisphere Language Battery (Batteria del Linguaggio dell'Emisfero Destro, BaLED) originally devised by Bryan. The normative study has been conducted with the recruitment of 440 healthy subjects. The battery of tests was not intended to be cognitively oriented, by providing evidence of the cognitive impairments underpinning verbal pragmatic deficits; on the contrary, it permits the detection of the presence/absence of impairments in processing the main pragmatic features of verbal communication traditionally associated with right hemisphere lesions. Thus, apart from being a clinical tool for diagnosing pragmatic impairments of verbal communication, the BaLED represents a useful initial battery of tests for clinical assessment and for selecting specific populations of neurological patients suitable for investigation in further experimental studies. PMID- 15877184 TI - A randomised, double-blind, dose-ranging study to evaluate efficacy and safety of three doses of botulinum toxin type A (Botox) for the treatment of spastic foot. AB - Botulinum toxin A (BTX) injections have been used successfully in the treatment of post-stroke foot spasticity, but the optimal dose-response relationship for selected muscles has yet to be established. The aim of this study was to outline beneficial and unwanted effects of three different doses of BTX in the treatment of spastic foot. In this randomised, double-blind, dose-ranging study, 45 spastic feet were randomly allocated to one of three groups, each of which was treated with a different dosage of BTX. The doses were decided on the basis of suggestions in the literature. Outcome measures (Modified Ashworth Scale, Medical Research Council Scale, gait assessment, presence of Achilles tendon clonus, Visual Analogue Scales for Gait Function and Pain, Adverse Effects scale) were applied at baseline, 4 weeks and 4 months after treatment. All the groups showed significant scales scores improvements after treatment with BTX. Group II (mean BTX total dose: 322 U) and Group III (mean BTX total dose: 540 U) showed a greater and more prolonged response than Group I (mean BTX total dose: 167 U). Group III showed the highest rate of adverse effects 4 weeks post-treatment. BTX injections constitute a useful and safe method of improving post-stroke foot spasticity, associated pain, gait speed and function. In particular, the medium BTX dosages (320 UI spread over 2-5 muscles) were found to be both safe and effective in producing long-lasting improvement of spastic foot dysfunction. PMID- 15877185 TI - Botulinum toxin type A for upper limb spasticity following stroke: an open-label study with individualised, flexible injection regimens. AB - Current antispastic medications are unsatisfactory for spasticity treatment, but botulinum toxin type A (BTX-A) shows promise as a new therapeutic option. This open-label, prospective study aimed to assess the effectiveness of BTX-A in improving functional mobility in the early post-stroke population using an individualised, flexible range of doses and targeted muscle groups. Twenty-one stroke patients (13 male, 8 female) were enrolled and injected with BTX-A (Botox, Allergan, mean dose: 255 U; range: 185-300) according to individual spasticity patterns. Assessments were made at baseline and weeks 2, 4, 6, 10 and 16 post treatment. Outcome measures comprised: Modified Ashworth Scale (MAS), finger flexion scale (Bhakta), MRC scale, Physician's Rating Scale (PRS), Nine Hole Peg Test (9HPT), Motor Assessment Scale, Clinical Global Impression (CGI), Global Assessment of Spasticity (GASS) and Visual Analogue Scale (VAS) for pain assessment. Statistically significant improvements in muscle tone as determined by the MAS were found in all areas (except arm) till week 16 (p<0.05). Finger positioning improved for the study duration, whilst muscle power increased only slightly in specific muscles. PRS revealed significant improvements to week 10 and slight improvement in 9HPT performance in selected patients was observed. Motor Assessment Scale results were statistically significant for arm, hand and advanced hand functions, although the overall functional benefit was mild. GASS and CGI results also showed improvement. Pain was present only in 11 patients and did not significantly improve following treatment. Individualised BTX-A injection regimens may be an effective, reversible and safe new treatment option for patients with spasticity. Nevertheless, functional improvement may be reached only in selected patients. PMID- 15877186 TI - Postpartum cerebellar infarction and haemolysis, elevated liver enzymes, low platelet (HELLP) syndrome. AB - Pregnancy is considered to be a hypercoagulable state per se with an increased risk for cerebrovascular events, however cerebellar infarction has been rarely described in pregnant women. A nulliparous pre-eclamptic woman at 25 weeks' gestation was submitted to an echocardiographic exam that showed an impaired cardiac structure and function. After 2 h, the patient underwent caesarean section for diagnosis of haemolysis, elevated liver enzymes, low platelet (HELLP) syndrome. Afterwards her platelet count raised, and eight days later she developed nystagmus, ataxia, dysmetria and motor deficit in the right limbs and sensory impairment in the right side of the face and in the left limbs. Cerebral magnetic resonance imaging (MRI) demonstrated a right cerebellar and median posterior bulbar infarction. Colour-coded sonography of cerebral vessels showed an occlusion of the right vertebral artery. Coagulation pattern analysis evidenced double heterozygosis of the methylenetetrahydrofolate reductase (MTHFR) gene and single mutation of the prothrombin gene. This case report gives evidence of the importance of considering the different risk factors involved in stroke occurrence during pregnancy. PMID- 15877187 TI - Pregnancy, patent foramen ovale and stroke: a case of pseudoperipheral facial palsy. AB - The pathogenetic role of patent foramen ovale (PFO) in embolic stroke and its prognostic and therapeutic implications have not yet been clearly defined. Nonetheless, recent availability of non-invasive diagnostic techniques, such as the transcranial Doppler (TCD), has increased the frequency with which this anomaly is diagnosed. Here we present the case of a young woman affected by post partum peripheral facial palsy: further exams disclosed not only its truncal ischaemic origin, but also, significantly, the presence of PFO, as well as of anticardiolipin antibodies (acL). Given the increased embolic risk in labouring women, this study highlights the importance of searching for PFO in case of a stroke during pregnancy. PMID- 15877188 TI - Hemifacial spasm due to a tentorial paramedian meningioma: a case report. AB - Hemifacial spasm (HFS) is a movement disorder characterised by involuntary paroxysmal facial movements that usually involve the orbicularis oculi and then spread to the other facial muscles. A microvascular compression and demyelination of the seventh nerve at its exit from the brain stem is considered to be the main aetiology of HFS. In addition to rare idiopathic (cryptogenetic) cases, others causes of HFS exist: tumours or vascular malformations have been described, of both the ipsilateral and contralateral cerebellopontine angle (CPA). However, space-occupying lesions in locations other than CPA are usually not thought to be responsible for HFS. Here we describe the case of a 45-year-old woman suffering from HFS, who dramatically improved after surgical removal of a tentorial paramedian meningioma. PMID- 15877189 TI - Dural arteriovenous fistulas with aggressive course: clinical and angiographic correlations in two patients. AB - Cranial dural arteriovenous fistulas (DAVFs) usually present with non-aggressive symptoms. We here report two patients who presented a peculiar clinical picture related to DAVFs, with focal neurological signs and haemorrhagic (case 1) or ischaemic lesions (case 2) respectively. The clinical and angiographic findings and putative pathophysiological mechanisms are discussed. PMID- 15877190 TI - Reversible choreoathetosis as the early onset of HIV-encephalopathy. PMID- 15877192 TI - The purchase of fruitfulness: assisted conception and reproductive disability in a seventeenth-century comedy. AB - The relationships between socioeconomic and biogenetic reproduction are always socially constructed but not always acknowledged. These relationships are examined as they apply to an instance of infertility and assisted reproduction presented in a seventeenth-century English play, Thomas Middleton's 1613 comedy, A Chaste Maid in Cheapside. Middleton's satirization of the effects of secrecy on the category of reproductive disability is analyzed and its applicability to our own time considered. The discussion is in four parts, focusing on: the attribution of disabled status to one member of the couple, the wife; the use of this attribution to protect the husband's reputation for sexual and reproductive health; the concealment of the nature of assisted reproduction; and the interests of the child conceived with such assistance. PMID- 15877193 TI - Looking in the mirror: images of abnormally developed infants. AB - Observational drawing provides a means of focusing on anomalous infant bodies. Time required by drawing connects the artist to the humanity of the subjects rather than to the deformities that make them, initially, frightening. PMID- 15877194 TI - Constructing prevention: Fetal Alcohol Syndrome and the problem of disability models. AB - Both the medical model and the social model of disability have substantial drawbacks for the project of creating better lives for people with disabilities; the first denies the value of difference and the effects of discrimination, and the second denies any place for prevention and cure. Using fictional and non fictional parental narratives of Fetal Alcohol Syndrome, this article argues that a third model--a morphological model of disability--can best help us think about respectfully and effectively intervening in disability. PMID- 15877195 TI - Mothers and models of disability. AB - Based on a qualitative anthropological study of American mothers of infants and young children newly diagnosed with disability, this essay examines how mothers understand their children and define disability in relation to publicly available discourses of disability and identity. In seeking to improve their children's opportunities in mainstream society, mothers appear to comply with the medical model. But over time and in the process of providing meaning to their experience, mothers retool models, drawing both on the social and minority group models' rejection of a problem-based definition of disability as inherently caused by impairment and on their own intimate engagement with impairment as an embodied experience. PMID- 15877196 TI - Reconsidering the social location of the medical model: an examination of disability in parenting literature. AB - This paper challenges the view that there is one medical model of disability monolithically and oppressively imposed on disabled people. Because the presence of disability may be ambiguous in any given case, multiple actors, lay and professional, may invoke particular medical models of disability and advance competing claims about an individual's disabilities and related needs. The literature for parents of disabled children is seen as a resource on which parents can draw in making claims about their children's disabilities and disability-related needs. Particular attention is given to the assumptions that this literature makes about the identity, resources and dispositions of the audience. Ultimately, these assumptions favor disability claims made by white, highly educated, upper-income parents. PMID- 15877197 TI - Textual abuse: Faulkner's Benjy. AB - William Faulkner's The Sound and the Fury has become a classic in literary history. Since its publication in 1929, it has sustained critical interest worldwide. Over time, analyses of this work have reflected shifting cultural perspectives of the inscribed human dynamics such as gender, race and sexuality. This paper contends that a similar critical development cannot be detected around the reception of the character of the "idiot," Benjy. Faulknerian scholarship, regardless of its place in time or trend, persists in conflating the dehumanized images of Benjy with the lived experience of disability, thus perpetuating oppressive disability prejudice and limiting the richness of the character's metaphoric potential. Acknowledging this critical lacuna, historicizing and theorizing Benjy's character from a disability perspective could lead to a deeper understanding of human experience. PMID- 15877198 TI - Braces, wheelchairs, and iron lungs: the paralyzed body and the machinery of rehabilitation in the polio epidemics. AB - The successful fund raising appeals of the March of Dimes employed images of cute crippled children standing on braces and forearm crutches, sitting in wheelchairs, or confined to iron lungs. Those who had to use these devices as a result of polio, however, were often stigmatized as cripples. American cultural antipathy to these assistive devices meant that polio survivors often had to overcome an emotional and psychological resistance to using them. Whatever their fears, polio survivors quickly discovered the functionality of braces and wheelchairs. By confronting the cultural stigma associated with these devices and in some sense embracing these mechanical "friends," polio survivors compensated for their paralyzed bodies and became active in the wider world of home, school and work. PMID- 15877199 TI - Cerebral ketone body metabolism. AB - Ketone bodies (KBs) are an important source of energy for the brain. During the neonatal period, they are also precursors for the synthesis of lipids (especially cholesterol) and amino acids. The rate of cerebral KB metabolism depends primarily on the concentration in blood; high concentrations occur during fasting and on a high-fat diet. Cerebral KB metabolism is also regulated by the permeability of the blood-brain barrier (BBB), which depends on the abundance of monocarboxylic acid transporters (MCT1). The BBB's permeability to KBs increases with fasting in humans. In rats, permeability increases during the suckling period, but human neonates have not been studied. Monocarboxylic acid transporters are also present in the plasma membranes of neurons and glia but their role in regulating KB metabolism is uncertain. Finally, the rate of cerebral KB metabolism depends on the activities of the relevant enzymes in brain. The activities vary with age in rats, but reliable results are not available for humans. Cerebral KB metabolism in humans differs from that in the rat in several respects. During fasting, for example, KBs supply more of the brain's energy in humans than in the rat. Conversely, KBs are probably used more extensively in the brain of suckling rats than in human neonates. These differences complicate the interpretation of rodent studies. Most patients with inborn errors of ketogenesis develop normally, suggesting that the only essential role for KBs is as an alternative fuel during illness or prolonged fasting. On the other hand, in HMG-CoA lyase deficiency, imaging generally shows asymptomatic white-matter abnormalities. The ability of KBs to act as an alternative fuel explains the effectiveness of the ketogenic diet in GLUT1 deficiency, but its effectiveness in epilepsy remains unexplained. PMID- 15877200 TI - Increased NO production in lysinuric protein intolerance. AB - Lysinuric protein intolerance (LPI) is a disorder of dibasic amino acid transport secondary to mutation of the SLC7A7 gene characterized by renal failure, pulmonary alveolar proteinosis, lupus-like autoimmune symptoms and usually increased plasma citrulline. In order to better understand the underlying mechanism, we studied the plasma and urinary nitrite/nitrate (NO2-/NO3-) concentrations in three LPI patients and the in vitro NO2- production in cultured fibroblasts. Our data show that NO3- levels are increased in the plasma of patients with LPI. Similarly, NO2- release in the medium of cultured fibroblasts was increased. On this basis, we hypothesize that some of the poorly understood clinical signs of LPI could be related to the activation of the NO-citrulline pathway. PMID- 15877201 TI - Hyperinsulinism in tyrosinaemia type I. AB - Tyrosinaemia type I (TT I) (McKusick 276700) is a heterogeneous disorder with a broad spectrum of clinical phenotypes. Although histological abnormalities of the pancreas are well recognized, there are only incidental reports of pancreatic dysfunction manifested as insulin-dependent diabetes mellitus. We report three subjects with TT I and acute liver dysfunction who had hyperinsulinism in early infancy. Hypoglycaemia persisted despite dietary treatment and one patient had inadequate lipolysis at the time of hypoglycaemia. All three patients were successfully treated with diazoxide (10 mg/kg per day) and chlorthiazide (35 mg/kg per day) and treatment was gradually withdrawn after 9, 13 and 34 months, respectively. The mechanism of pancreatic dysfunction in TT I is unknown but may be related to the toxic metabolites that accumulate in this condition. We conclude that hyperinsulinism is not a rare complication in TT I. In patients with persistent hypoglycaemia, C-peptide should always be measured. Treatment with diazoxide and chlorthiazide is highly effective, appears to be safe, and does not need to be continued lifelong. PMID- 15877202 TI - Hyperchylomicronaemia due to lipoprotein lipase deficiency as a cause of false positive newborn screening for biotinidase deficiency. AB - Two cases of molecular genetically proven lipoprotein lipase deficiency are reported. Both patients were detected owing to a false-positive neonatal screening test for biotinidase deficiency. We conclude that both the fluorimetric and the colorimetric screening tests for biotinidase deficiency used with dried blood samples are affected by severe hyperchylomicronaemia and that, most probably, severe plasma turbidity interferes with the assay. PMID- 15877203 TI - L-carnitine and exercise tolerance in medium-chain acyl-coenzyme A dehydrogenase (MCAD) deficiency: a pilot study. AB - Skeletal muscle function may be impaired in patients with medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, but the value of L-carnitine in their long-term management is not clear. This study was designed as a pilot to examine the effects of L-carnitine on exercise tolerance in patients with MCAD deficiency. Four clinically asymoptomatic MCAD-deficient patients, aged 8 to 20 years, were studied. Incremental ramp exercise tests were carried out before and after 4 weeks' treatment with oral L-carnitine (100 mg/kg per day). During exercise without L-carnitine supplementation, plasma carnitine concentrations fell, associated with an increased excretion of urinary acylcarnitines, notably acetylcarnitine, hexanoylcarnitine and octanoylcarnitine. L-carnitine treatment prevented this fall in plasma carnitine and resulted in greater increases in excretion of acylcarnitines. All four patients showed biologically significant improvement in peak oxygen uptake (peak VO2, 18-32% improvement), VO2 at a heart rate of 170 beats/min (15-23% improvement), VO2 at anaerobic threshold (27-42% improvement), and/or oxygen pulse (10-32% improvement). Exercise tolerance in MCAD-deficient patients may be improved by short-term L-carnitine supplementation. This may be the direct result of improved intramitochondrial homeostasis induced by L-carnitine in removing accumulating acyl moieties. PMID- 15877204 TI - Hepatocellular carcinoma in glycogen storage disease type Ia: a case series. AB - We present a series of 8 patients (6 males, 2 females) with hepatocellular carcinoma (HCC) and glycogen storage disease type Ia (GSD Ia). In this group, the age at which treatment was initiated ranged from birth to 39 years (mean 9.9 years). All patients but one were noncompliant with treatment. Hepatic masses were first detected at an age range of 13-45 years (mean 28.1 years). Age at diagnosis of HCC ranged from 19 to 49 years (mean 36.9 years). Duration between the diagnosis of liver adenomas and the diagnosis of HCC ranged from 0 to 28 years (mean 8.8 years, SD = 11.5). Two patients had positive hepatitis serologies (one hepatitis B, one hepatitis C). Alpha-fetoprotein (AFP) was normal in 6 of the 8 patients. Carcinoembryonic antigen (CEA) was normal in the 5 patients in which it was measured. Current guidelines recommend abdominal ultrasonography with AFP and CEA levels every 3 months once patients develop hepatic lesions. Abdominal CT or MRI is advised when the lesions are large or poorly defined or are growing larger. We question the reliability of AFP and CEA as markers for HCC in GSD Ia. Aggressive interventional management of masses with rapid growth or poorly defined margins may be necessary to prevent the development of HCC in this patient population. PMID- 15877205 TI - Galactosaemia: early treatment with an elemental formula. AB - Classical galactosaemia is caused by deficient galactose-1-phosphate uridyl transferase activity, and is treated by dietary galactose restriction. Despite dietary treatment, long-term outcomes have not been uniformly favourable. Late complications may include speech abnormalities, ataxia, cognitive impairment, growth delay, bone alterations and ovarian failure. We report an infant whose erythrocyte galactose 1-phosphate (gal-1-P) levels remained well above the treatment range on a low-galactose (soy) formula. Once she was begun on an elemental formula (galactose-free), gal-1-P levels decreased rapidly to within the treatment range. Urine galactitol levels decreased on the elemental formula but were within published treatment ranges despite treatment changes. These did not correlate with the gal-1-P levels. This case suggests further study be considered to determine whether a truly galactose-free diet in infancy could alter the long-term prognosis of classical galactosaemia. PMID- 15877206 TI - A newborn with severe liver failure, cardiomyopathy and transaldolase deficiency. AB - This paper describes the second patient found to be affected with a deficiency of transaldolase (TALDO1; EC 2.2.1.2). Clinically, this patient presented in the neonatal period with several signs of severe liver failure: severe coagulopathy, low serum protein, elevated blood ammonia, and hypoglycaemia. She had generalized oedema, moderate muscular hypotonia, and dysmorphic signs. Liver size was decreased, and the spleen was moderately enlarged. There was severe cardiomegaly. The clinical course was characterized by intractable liver failure and progressive myocardial hypertrophy. The child died at the age of 18 days from respiratory failure. In urine, elevations of erythritol, arabitol and ribitol were found, suggesting a deficiency of transaldolase. Enzyme studies in cultured fibroblasts showed undetectable transaldolase activity. DNA sequence analysis of the TALDO1 gene showed a homozygous missense mutation (575G>A), resulting in an amino acid alteration at position 192 (arginine to histidine, R192H). This amino acid is part of the catalytic site of the transaldolase protein. Discovery of this second patient affected with transaldolase deficiency and liver failure suggests that this disorder has a heterogeneous clinical presentation with highly variable severity. PMID- 15877207 TI - Selective antibody immune deficiency in a patient with Smith-Lemli-Opitz syndrome. AB - Smith-Lemli-Opitz syndrome is a rare autosomal recessive disorder characterized by multiple congenital anomalies and various degrees of cognitive deficits. This condition results from a deficiency of 7-dehydrocholesterol reductase, a critical step in cholesterol biosynthesis. Children with Smith-Lemli-Opitz syndrome have frequent infections, particularly of the respiratory tract. Immunodeficiency, however, is not recognized as a part of this metabolic condition. Frequent infections are usually attributed to a decreased patient mobility and reduced respiratory effort secondary to muscular hypotonia and mental retardation, which are often present in affected individuals. We describe a patient with Smith-Lemli Opitz syndrome and recurrent respiratory infections who was found to have a selective antibody deficiency. The immunological diagnosis was based on an absent immune response to Pneumovax. She also had no immunological response to hepatitis B vaccine and was unable to break down red cells with isoagglutinin B. Therapy with intravenous IgG (IVIG) was initiated. Infections were less severe, although they still occurred in a high frequency after initiation of the IVIG therapy. This finding prompts the need for a higher index of suspicion for an underlying immune deficiency in patients with Smith-Lemli-Opitz syndrome who present with recurrent and chronic infections. Early recognition and appropriate therapeutic interventions may decrease the severity of infections, prevent potentially fatal infections, and eventually improve the quality of life in these patients. PMID- 15877208 TI - Keratan sulphate levels in mucopolysaccharidoses and mucolipidoses. AB - The mucopolysaccharidoses (MPS) is characterized by accumulation of glycosaminoglycans (GAGs), and mucolipidosis (ML) by accumulation of GAGs and sphingolipids. Each type of MPS accumulates specific GAGs. The lysosomal enzymes N-acetylgalactosamine-6-sulphate sulphatase and beta-galactosidase involve the stepwise degradation of keratan sulphate (KS). Deficiency of these enzymes results in elevation of KS levels in the body fluids and in tissues, leading to MPS IV disease. In this study, we evaluated blood and urine KS levels in types of MPS and ML other than MPS IV. Eighty-five plasma samples came from MPS I (n = 18), MPS II (n = 28), MPS III (n = 20), MPS VI (n = 3), MPS VII (n = 5) and ML (n = 11) patients while 127 urine samples came from MPS I (n = 34), MPS II (n = 34), MPS III (n = 32), MPS VI (n = 7), MPS VII (n = 9) and ML (n = 11) patients. KS levels were determined using the ELISA method. Plasma KS levels varied with age in both control and patient populations. In all age groups, the mean values of plasma KS in MPS and ML patients were significantly higher than those in the age matched controls. Plasma KS values in four newborn patients were above the mean + 2SD of the age-matched controls (mean, 41 ng/ml). Overall, 85.9% of individual values in non-type IV MPS and ML patients were above the mean + 2SD of the age matched controls. For urine KS levels, 24.4% of individual values in patients were above the mean + 2SD of the age-matched controls. In conclusion, KS in blood is elevated in each type of non-type IV MPS examined, in contrast to the conventional understanding. This finding suggests that measurement of KS level provides a new diagnostic biomarker in a wide variety of mucopolysaccharidoses and mucolipidoses in addition to MPS IV. PMID- 15877209 TI - Acid sphingomyelinase deficiency. Phenotype variability with prevalence of intermediate phenotype in a series of twenty-five Czech and Slovak patients. A multi-approach study. AB - A multi-approach study in a series of 25 Czech and Slovak patients with acid sphingomyelinase deficiency revealed a broad phenotypic variability within Niemann-Pick disease types A and B. The clinical manifestation of only 9 patients fulfilled the historical classification: 5 with the rapidly progressive neurovisceral infantile type A and 4 with a slowly progressive visceral type B. Sixteen patients (64%) represented a hitherto scarcely documented 'intermediate type' (IT). Twelve patients showed a protracted neurovisceral course with overt or mild neurological symptoms, three a rapidly progressing fatal visceral affection with rudimentary neurological lesion. One patient died early from a severe visceral disease. The genotype in our patients was represented by 4 frameshift and 14 missense mutations. Six were novel (G166R, R228H, A241V, D251E, D278A, A595fsX601). The Q292K mutation (homoallelic, heteroallelic) was strongly associated with a protracted neurovisceral phenotype (10 of 12 cases). The sphingomyelin loading test in living fibroblasts resulted in total degradation from less than 2% in classical type A to 70-80% in classical type B. In the IT group it ranged from 5% to 49% in a 24 h chase. The liver storage showed three patterns: diffuse, zonal (centrolobular), and discrete submicroscopic. Our series showed a notable variability in both the neurological and visceral lesions as well as in their proportionality and synchrony, and demonstrates a continuum between the historical 'A' and 'B' phenotypes of ASM deficiency. This points to a broad phenotypic potential of ASM deficiency, suggesting the existence of still unknown factors independently controlling the storage level in the visceral and neuronal compartments. This report highlights the important position of the IT in the ASM deficiency phenotype classification. We define IT as a cluster of variants combining clinical features of both the classical types. The protracted neuronopathic variant with overt, borderline or subclinical neurology prevails and is important in view of future enzyme replacement therapy. It appears more common in central Europe. The visceral, rapidly progressing early fatal type has been recognized rarely so far. PMID- 15877210 TI - Consanguineous 3-methylcrotonyl-CoA carboxylase deficiency: early-onset necrotizing encephalopathy with lethal outcome. AB - A patient with a severe neonatal variant of 3-methylcrotonyl-CoA carboxylase (MCC) deficiency is reported. The first child of healthy consanguineous Turkish parents presented on the second day of life with dehydration, cyanosis, no sucking, generalized muscular hypotonia, encephalopathy, respiratory depression requiring mechanic ventilation, macrocephaly, severe acidosis and hypoglycaemia. Elevated C5-OH-carnitine in dried blood spot by tandem MS and elevated urinary excretion of 3-hydroxyisovaleric acid and 3-methylcrotonylglycine suggested MCC deficiency, confirmed by enzyme analysis in cultured fibroblasts. Cerebral ultrasonography and cranial CT findings revealed progressive changes such as disseminated encephalomalacia, cystic changes, ventricular dilatation and cerebral atrophy. Treatment with high-dose biotin and protein-restricted diet was ineffective and the patient died at the age of 33 days with progressive neurological deterioration. Mutation analysis revealed a homozygous mutation in the splice acceptor site of intron 15 in the MCC beta-subunit. Early-onset severe necrotizing encephalopathy should be included in the differential diagnosis of isolated MCC deficiency. PMID- 15877211 TI - Two novel mutations in mitochondrial acetoacetyl-CoA thiolase deficiency. AB - We report a new patient with acetoacetyl-CoA thiolase deficiency in whom we found two new missense mutations. PMID- 15877212 TI - Ornithine transcarbamylase deficiency in pregnancy. AB - Women heterozygous for mutations at the ornithine transcarbamylase (OTC) locus may be at risk for hyperammonaemia and its untoward effects including coma and death in the postpartum period. We present the case of a pregnant woman heterozygous for OTC deficiency (McKusick 311250) whose past medical history was significant for two prior pregnancies complicated by postpartum hyperammonaemic coma. In the index pregnancy, increased levels of serum ammonium were noted during labour. Postpartum hyperammonaemia was averted by administration of oral sodium benzoate. Our experience demonstrates that in women at risk, perilous hyperammonaemia can be prevented through appropriate medical management. PMID- 15877213 TI - N-carbamylglutamate protects patients with decompensated propionic aciduria from hyperammonaemia. AB - In patients with propionic aciduria, the accumulating metabolite propionyl-CoA causes a disturbance of the urea cycle via the inhibition of N-acetylglutamate synthesis. Lack of this allosteric activator results in an inhibition of carbamoylphosphate synthase (CPS). This finally leads to hyperammonaemia. In two patients with decompensated propionic aciduria the CPS activator carbamylglutamate was tested for its ability to antagonize the propionyl-CoA associated hyperammonaemia. Oral carbamyl glutamate administration resulted in a significant increase in ammonia detoxification and could avoid further dialysis therapy. Safe, fast and easy to administer, carbamyl glutamate improves the acute therapy of decompensated propionic aciduria by increasing ammonia detoxification and avoiding hyperammonaemia. PMID- 15877214 TI - [Stereotactic radiation therapy for liver metastases]. AB - Advances in radiation oncology made possible the locally curative treatment approach of stereotactically guided radiation therapy of liver tumours. Results of this highly focussed therapy were published by only a few centres. The radiation dose is delivered in one or a few fractions with high single doses. All published results show high local tumour control with low treatment morbidity. The treatment results of this noninvasive technique are similar to those of minimally invasive ablative therapies. Our own and other published data are summarized and discussed. The long-term results of this technique are currently being evaluated in a prospective multicentre trial. PMID- 15877215 TI - The adiponectin gene SNP+276G>T associates with early-onset coronary artery disease and with lower levels of adiponectin in younger coronary artery disease patients (age G and +276 G>T) of the adiponectin gene and CAD, looking also into the possible influence of these SNPs on adiponectin plasma levels. The SNPs were analysed in a first cohort of 595 subjects, 325 with CAD and 270 matched controls. We observed a significant association (p<0.001) between the SNP +276G>T in the adiponectin gene and CAD. In multivariate analysis, carriers of the +276G>T SNP had an odds ratio (OR) for CAD of 4.99 (p<0.0007). A strong interaction between the +276G>T SNP and age was also present (OR, 1.03; p<0.0001). The increase in CAD risk was most evident among individuals with early onset CAD (age T SNP was analysed in a second cohort of CAD and controls. The difference between CAD and controls in the +276G>T SNP frequencies showed a similar trend as before, although not significant. The combination of the two cohorts (1,046 subjects: 580 CAD and 466 controls) showed a statistically significant association, particularly in CAD subjects with early-onset of disease. In addition, we confirmed that in younger CAD subjects the SNP was a significant determinant of lower levels of adiponectin. In view of these results, it could be speculated that the adiponectin gene variant, or a mutation in linkage with it, determines lower adiponectin gene expression, causing in turn an increased risk to develop insulin resistance, atherosclerosis and cardiovascular disease. The significant association of the adiponectin gene in subjects with early-onset CAD also suggests that that genetic factors for late-onset diseases may exert a greater influence in younger persons, when other risk factors are not as prevalent as in older age groups. PMID- 15877216 TI - Angiotensin II type 1 receptor inhibition markedly improves the blood perfusion, oxygen tension and first phase of glucose-stimulated insulin secretion in revascularised syngeneic mouse islet grafts. AB - AIMS/HYPOTHESIS: We recently found evidence of an angiotensin-generating system in pancreatic islets. The present study investigated the effect of endogenously produced angiotensin II on microcirculation and function in transplanted islets. MATERIALS AND METHODS: Losartan, an angiotensin II type 1 receptor inhibitor, was administered either acute intravenously to mice with 4-week-old islet renal subcapsular transplants, or added to the drinking water for the final 14 days or throughout the 4-week post-transplantation period. The graft-bearing kidney was, in some cases, dissected out and perfused in vitro to evaluate the effect of angiotensin II and losartan on glucose-stimulated insulin release from the grafts. RESULTS: Losartan treatment throughout the 4-week post-transplantation period had negative effects on islet revascularisation as well as on islet graft insulin release. However, administration of losartan, either intravenously or orally, after the formation of a new vascular network, improved islet graft blood perfusion. PO2 in the islet transplants was also effectively improved by the losartan treatment. Graft perfusion experiments showed a markedly better first phase of glucose-stimulated insulin release in transplanted islets when exposed to losartan. In contrast, acute administration of angiotensin II decreased islet graft blood flow, PO2 and glucose-stimulated insulin release. CONCLUSIONS/INTERPRETATION: This study shows that inhibition of the islet reninangiotensin system may be a feasible strategy to increase the blood perfusion, PO2 and function within islet grafts. Such treatment should not be initiated, however, before the islet vascular system has been formed. PMID- 15877217 TI - Preliminary remarks on a prospective multicenter study of the Repicci minimally invasive unicondylar knee replacement. AB - Prospective multicentric study on unicondylar knee arthroplasties using the minimally invasive method described by Repicci. Three hundred and seventy nine unicondylar knee arthroplasties in patients with an average age of 66 years were examined preoperatively and 40 weeks postoperatively. In 98.4% of the cases the medial compartment was replaced. The examination was documented using the Knee Society Score (KSS). Radiological examination was done using films of the knee in two planes in standing position and a long leg axis film. The mean KSS increased significantly (P<0.005) from 100.1 (SD 21.7) before surgery to 179.1 (SD 24.0) postoperatively. Ninety-five of the patients were subjectively satisfied with the procedure. Using correlation analysis, we found that osteoarthritic changes of the patella had a highly significant (p=0.001) influence on the KSS results. Patient's age and the pain subscale of the KSS also had a highly significant (p=0.01) influence on the KSS result. Short-term results of the Repicci unicondylar arthroplasty using the minimally invasive technique are encouraging in terms of patient satisfaction. It became apparent that osteoarthritic changes of the patella, flexion contractures and extension deficits have a considerable influence on the overall result. PMID- 15877218 TI - Thermal effects after anterior cruciate ligament shrinkage using radiofrequency technology: a porcine cadaver study. AB - The effects of thermal energy on an anterior cruciate ligament (ACL) are unknown. This study was undertaken to know the immediate and direct effects of thermocoagulation on normal ACL. Thermocoagulation was generated by monopolar radiofrequency (RF) at 67 degrees C and 40 W of energy and applied with use of a commercial jig in a porcine cadaveric model, and biomechanical and histological changes were evaluated immediately. Thermocoagulation caused significant shrinkage of ACL without immediate effects on ligament biomechanical properties, including stiffness and maximal failure force. However, histological analysis demonstrated a decrease in number of dense elongated cells and blood vessels within the epiligamentous tissue, loss of areolar spaces, decrement in the periodicity of collagen waveform, and shortening and pyknotic nuclear changes of cells within the fascicular region. RF-generated thermocoagulation resulted in ACL shortening with significant changes on histological findings but not on initial biomechanical properties. PMID- 15877219 TI - Eccentric training in chronic painful impingement syndrome of the shoulder: results of a pilot study. AB - Treatment with painful eccentric muscle training has been demonstrated to give good clinical results in patients with chronic Achilles tendinosis. The pain mechanisms in chronic painful shoulder impingement syndrome have not been scientifically clarified, but the histological changes found in the supraspinatus tendon have similarities with the findings in Achilles tendinosis. In this pilot study, nine patients (five females and four males, mean age 54 years) with a long duration of shoulder pain (mean 41 months), diagnosed as having shoulder impingement syndrome and on the waiting list for surgical treatment (mean 13 months), were included. Patients with arthrosis in the acromio-clavicular joint or with large calcifications causing mechanical impingement during horizontal shoulder abduction were not included. We prospectively studied the effects of a specially designed painful eccentric training programme for the supraspintus and deltoideus muscles (3 x 15 reps, 2 times/day, 7 days/week, for 12 weeks). The patients evaluated the amount of shoulder pain during horizontal shoulder activity on a visual analogue scale (VAS), and satisfaction with treatment. Constant score was assessed. After 12 weeks of treatment, five patients were satisfied with treatment, their mean VAS had decreased (62-18, P<0.05), and their mean Constant score had increased (65-80, P<0.05). At 52-week follow-up, the same five patients were still satisfied (had withdrawn from the waiting list for surgery), and their mean VAS and Constant score were 31 and 81, respectively. Among the satisfied patients, two had a partial suprasinatus tendon rupture, and three had a Type 3 shaped acromion. In conclusion, the material in this study is small and the follow-up is short, but it seems that although there is a long duration of pain, together with bone and tendon abnormalities, painful eccentric supraspinatus and deltoideus training might be effective. The findings motivate further studies. PMID- 15877220 TI - Evaluation of the second-generation meniscus arrow in the fixation of bucket handle tears in the vascular area of the meniscus. A prospective study of 20 patients with a mean follow-up of 26 months. AB - The purpose of this prospective study was to evaluate whether the shorter degradation time of the second-generation Meniscus Arrow would be adequate to allow meniscal healing and whether equally good results could be achieved compared to the first generation implant. We also wanted to assess whether some of the implant related complications could be avoided with the altered absorption time. A total of 20 patients were included in this study. The mean follow-up was 26 months. The average Lysholm score at the final follow-up was 90. There were 13 second-look arthroscopies performed during the follow-up and in them two failures of the repaired menisci to heal were seen. Seven patients were found with some degree of chondral irritation of the femoral condyle corresponding to the repair site. The second generation Meniscus Arrow seems to give results comparable to the first generation Meniscus Arrow. The potential complications caused by the implant reported in the earlier literature using the first generation arrow are possible also using this implant. In seven of the 13 second-look arthroscopies some degree of chondral irritation corresponding to the repair site was seen. The clinical relevance of these lesions remains to be seen. PMID- 15877221 TI - Knee extensors kinetic chain training in anterior cruciate ligament deficiency. AB - Open kinetic chain (OKC) knee extensor resistance training has lost favour in rehabilitation of patients with knee ACLD due to concerns that this exercise is harmful to the remaining portion of the ACL and its secondary stabilizers, and will be less effective in improving function. In this randomized, single-blind clinical trial, closed and OKC knee extensor training were compared for their effects on knee laxity and function in patients with ACLD knees. Sixty-four patients with a diagnosis of knee ACLD (49 M, 15 F; mean age=30 years) were measured for knee laxity, using a ligament arthrometer, and function with the Hughston Clinic knee self-assessment questionnaire and maximal effort single leg jump testing. Between the above tests and identical tests carried out 6 weeks later, subjects trained using either open or closed kinetic chain resistance of their knee and hip extensors as part of formal physical therapy sessions three times per week for 6 weeks. The groups exhibited no statistically significant differences (p<0.083) in outcome. These results indicate that knee extensor OKC training, as used in this study, can be safely applied to patients with knee ACL deficiency, and shows no superiority to CKC training. PMID- 15877222 TI - ICP-MS multielemental determination of metals potentially released from dental implants and articular prostheses in human biological fluids. AB - A sector field high-resolution (HR)-ICP-MS and an octapole reaction system (ORS) ICP-MS have been compared for the simultaneous determination of traces of metals (Ti, V, Cr, Co, Ni, and Mo) released from dental implants and articular prostheses in human biological fluids. Optimum sample treatments were evaluated to minimize matrix effects in urine and whole blood. Urine samples were diluted tenfold with ultrapure water, whereas whole blood samples were digested with high purity nitric acid and hydrogen peroxide and finally diluted tenfold with ultrapure water. In both matrices, internal standardization (Ga and Y) was employed to avoid potential matrix interferences and ICP-MS signal drift. Spectral interferences arising from the plasma gases or the major components of urine and whole blood were identified by (HR)-ICP-MS at 3,000 resolving power. The capabilities of (HR)-ICP-MS and (ORS)-ICP-MS for the removal of such spectral interferences were evaluated and compared. Results indicate that polyatomic interferences, which hamper the determination of such metallic elements in these biological samples, could be overcome by using a resolving power of 3,000. Using (ORS)-ICP-MS, all those elements could be quantified except Ti and V (due to the polyatomic ions 31P16O and 35Cl16O, respectively). The accuracy of the proposed methodologies by (HR)- and (ORS)-ICP-MS was checked against two reference materials. Good agreement between the given values and the concentrations obtained for all the analytes under scrutiny was found except for Ti and V when analyzed by (ORS)-ICP-MS. PMID- 15877223 TI - Proteolysis and partial dissolution of calcium oxalate: a comparative, morphological study of urinary crystals from black and white subjects. AB - Crystal adherence to the renal epithelium is widely regarded as a probable mechanism of stone formation. Intracrystalline proteins may provide access to the core of urinary crystals and thereby facilitate the dismantling of these crystals after their attachment to and phagocytosis by renal epithelial cells. The present study investigated the role of proteolysis and limited dissolution of urinary calcium oxalate (CaOx) crystals in South Africa's white and black populations with a view to understanding the remarkably low stone incidence in the black population compared with the whites. CaOx crystals were precipitated from filtered urine or ultrafiltered urine dosed with an intracrystalline protein, urinary prothrombin fragment 1 (UPTF1), isolated from white and black subjects. The crystals were fractured and subjected to proteolysis and/or limited dissolution before examination using field emission scanning electron microscopy (FESEM). FESEM data showed that CaOx crystals from white and black subjects were eroded by treatment with proteases. Cathepsin D caused the most significant crystal erosion, and more noticeable degradation of CaOx monohydrate (COM) crystals compared to CaOx dihydrate (COD). Limited dissolution studies showed the unique ultrastructures and fragmentation processes of COM and COD crystals. COM crystals appeared to be more susceptible to degradation and dissolution than CODs. Since COMs are predominant in blacks, compared with COD crystals from whites, it is speculated that the lower stone rate amongst South African blacks might be attributed partly to their more efficient destruction of retained COM crystals. PMID- 15877224 TI - A novel antioxidant agent caffeic acid phenethyl ester prevents shock wave induced renal tubular oxidative stress. AB - The aim of this study was to evaluate the effects of the novel free radical scavenger caffeic acid phenethyl ester (CAPE) on extracorporeal shock wave lithotripsy (ESWL) induced renal impairment. The study was performed using 30 rabbits which were divided into two groups, each exposed to 3,000 shock waves at 18 kV: (1) control group, (2) ESWL+CAPE treated group. Malodialdehyde (MDA), urine N-acetyl-beta-glucosaminidase (NAG) activity, uric acid and white cell counts were used as markers of oxidative stress. Following shock wave exposure there was a significant rise in MDA, NAG and uric acid and white cell counts. CAPE reduced the rise in MDA, NAG, uric acid and white cell counts. Thus CAPE treatment to a great extent prevented the induction of these renal changes. Our results suggest that the antioxidant capacity of the kidney tissue was reduced after ESWL treatment and that the tissue was exposed to oxidant stress. We conclude that CAPE treatment provided significant protection against ESWL induced free radical damage. PMID- 15877225 TI - Spinal epithelioid hemangioendothelioma with epithelioid angiosarcomatous areas. AB - Epithelioid hemangioendothelioma is a distinctive vascular tumor rarely involving bones. We report a case of epithelioid hemangioendothelioma in the cervical spine with angiosarcomatous areas. A 50-year-old female presented with dizziness while walking. A mixed sclerotic and osteolytic or destructive lesion was radiographically disclosed in her upper cervical vertebrae. Histologically, a laminectomy specimen exhibited areas of ordinary epithelioid hemangioendothelioma together with foci of more atypical epithelioid cell proliferation, closely resembling epithelioid angiosarcoma. This phenomenon has been exceptionally described in cases with skeletal lesions, which are typically of low-grade malignancy. PMID- 15877226 TI - O-(2-[18F]fluoroethyl)-L-tyrosine PET for monitoring the effects of convection enhanced delivery of paclitaxel in patients with recurrent glioblastoma. AB - PURPOSE: Convection-enhanced delivery (CED) of paclitaxel is a new locoregional approach for patients with recurrent glioblastoma. The aim of this study was to evaluate O-(2-[(18)F]fluoroethyl)-L-tyrosine (FET) positron emission tomography (PET) in monitoring the effects of this type of direct drug delivery. METHODS: Eight patients with recurrent glioblastoma underwent CED of paclitaxel, which was infused over stereotactically placed catheters into the tumour. FET PET and MRI were performed before and 4 weeks after therapy and then at 3-month intervals to document follow-up. For quantitative evaluation, SUV(max)(tumour)/SUV(mean)(background) ratios were calculated. RESULTS: At baseline all tumours showed gadolinium enhancement and high FET uptake (SUV(max)/BG 3.2+/-0.8). Four weeks after CED, a statistically significant decrease in FET uptake was seen (SUV(max)/BG-17%; p<0.01). During follow-up, no recurrence was observed within the CED area. Two out of eight patients with extended tumours died 4 and 5 months after treatment, most probably from local complications. Temporarily stable disease with stable FET uptake was observed in six of eight patients; this was followed by progression and increasing FET uptake ratios (+46%) distant from the CED area in five of the six patients 3-13 months after CED. One patient still presents stable FET uptake 10 months after CED. MRI showed unchanged/increasing contrast enhancement and oedema without ability to reliably assess disease progression. CONCLUSION: FET PET is a valuable tool in monitoring the effects of CED of paclitaxel. In long-term follow-up, stable or decreasing FET uptake, even in contrast-enhancing lesions, is suggestive of reactive changes, whereas increasing ratios appear always to be indicative of recurrence. Therefore, FET PET is more reliable than MRI in differentiating stable disease from tumour regrowth. PMID- 15877227 TI - Efficiency of 18F-FDG and 99mTc-depreotide SPECT in the diagnosis of malignancy of solitary pulmonary nodules. AB - PURPOSE: The purpose of the study was to compare ( 18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) and ( 99m)Tc-depreotide single-photon emission computed tomography (SPECT) in the diagnosis of malignancy of solitary pulmonary nodules (SPNs). METHODS: Twenty-eight patients without any history of cancer and presenting an SPN (0.8-3 cm in size) underwent FDG PET and depreotide SPECT. Depreotide SPECT and FDG PET were performed on a double-head gamma camera and a dedicated PET scanner respectively. Twenty-five out of 28 lesions were removed by thoracotomy or assessed by biopsy (n=1) and histologically examined. A strategy of serial CT scanning was adopted in the three remaining patients. RESULTS: Histological findings revealed 18 malignant nodules and seven benign lesions. Stability over a 2-year period indicated a benign process in the remaining three cases. Both techniques yielded true positive results in 15 of the 18 cancers. FDG PET identified two additional adenocarcinomas not detected by depreotide SPECT. A carcinoid tumour not visualised on FDG PET was identified by depreotide SPECT. Seven of the ten benign lesions did not reveal tracer uptake on either depreotide SPECT or FDG PET. Both techniques showed false positive results for the same two lesions. One more false positive was seen on FDG PET. FDG PET and depreotide SPECT had a sensitivity of 94.4% and 88.9% respectively; this difference was not significant. In our experience, depreotide SPECT and FDG PET are equally sensitive (92.3%) for large (>1.5 cm) and equally specific (85.7%) for small (up to 1.5 cm) SPNs suspicious for malignancy. CONCLUSION: This study showed( 18)F FDG PET to be more sensitive than ( 99m)Tc-depreotide SPECT in the diagnosis of malignancy of SPNs. However, the combination of both techniques may provide additional accuracy. PMID- 15877228 TI - Tumor immunity: a balancing act between T cell activation, macrophage activation and tumor-induced immune suppression. AB - The mouse 4T1 mammary carcinoma is a BALB/c-derived tumor that spontaneously metastasizes and induces immune suppression. Although >95% of wild type BALB/c mice die from metastatic 4T1 tumor even if the primary mammary tumor is surgically removed, >65% of BALB/c mice with a deleted Signal Transducer Activator of Transcription 6 (STAT6) gene survive post-surgery. STAT6-deficiency also confers enhanced immunity against spontaneously developing breast cancer since NeuT+/- mice that are STAT6-deficient develop mammary tumors later and survive longer than NeuT+/- mice that are STAT6-competent. Rejection of metastastic disease and survival of STAT6-deficient mice after removal of primary tumor involve three mechanisms: (1) The generation of M1 type macrophages that produce nitric oxide and are tumoricidal; (2) A decrease to normal in the elevated levels of myeloid suppressor cells that accumulate during primary tumor growth; and (3) CD8+ tumor-specific T lymphocytes. STAT6-deficient, but not wild type BALB/c, mice generate nitric oxide producing macrophages because they lack the STAT6 transcription factor which is necessary for signaling through the type 2 IL-4Ralpha complex, and which induces the production of arginase instead of nitric oxide. PMID- 15877229 TI - Identification of biliary metabolites of ifosfamide using 31P magnetic resonance spectroscopy and mass spectrometry. AB - Biliary excretion is a significant component in the metabolism of many drugs, but remains difficult to detect and characterise non-invasively. A previous publication recently described the detection of metabolites of ifosfamide in gall bladder in a guinea pig model using in vivo 1H-decoupled 31P 3-D magnetic resonance spectroscopic imaging and a clinical 1.5 T MR scanner.. Here high resolution 31P magnetic resonance spectroscopy (MRS) of extracted bile identifies peaks as parent ifosfamide (1.19+/-1.47 mM; mean+/-sd), carboxyifosfamide (2.04+/ 1.04 mM) and a major contribution from a previously unreported peak at 16.0 ppm (4.05+/-2.38 mM). The unknown resonance was identified using liquid chromatography-mass spectrometry (LCMS) as the glutathione conjugate of ifosfamide (MW=531). This was confirmed by analysing products from the reaction of glutathione with ifosfamide using LCMS and MRS. These results demonstrate how combined in vivo and analytical MRS, together with mass spectrometry, can help identify visceral routes of drug metabolism, thereby aiding understanding of +/ drug disposition and mechanisms of action and toxicity. In particular, the distribution of ifosfamide and its metabolites into bile may be related to oxazophosphorine-related cholecystitis reported in patients. PMID- 15877230 TI - Effect of NQO1 induction on the antitumor activity of RH1 in human tumors in vitro and in vivo. AB - NQO1 is a reductive enzyme that is important for the activation of many bioreductive agents and is a target for an enzyme-directed approach to cancer therapy. It can be selectively induced in many tumor types by a number of compounds including dimethyl fumarate and sulforaphane. Mitomycin C is a bioreductive agent that is used clinically for treatment of solid tumors. RH1 (2,5-diaziridinyl-3-(hydroxymethyl)- 6-methyl-1,4-benzoquinone) is a new bioreductive agent currently in clinical trials. We have shown previously that induction of NQO1 can enhance the antitumor activity of mitomycin C in tumor cells in vitro and in vivo. As RH1 is activated selectively by NQO1 while mitomycin C is activated by many reductive enzymes, we investigated whether induction of NQO1 would produce a greater enhancement of the antitumor activity of RH1 compared with mitomycin C. HCT116 human colon cancer cells and T47D human breast cancer cells were incubated with or without dimethyl fumarate or sulforaphane followed by mitomycin C or RH1 treatment, and cytotoxic activity was measured by a clonogenic (HCT116) or MTT assay (T47D). Dimethyl fumarate and sulforaphane treatment increased NQO1 activity by 1.4- to 2.8-fold and resulted in a significant enhancement of the antitumor activity of mitomycin C, but not of RH1. This appeared to be due to the presence of a sufficient constitutive level of NQO1 activity in the tumor cells to fully activate the RH1. Mice were implanted with HL60 human promyelocytic leukemia cells, which have low levels of NQO1 activity. The mice were fed control or dimethyl fumarate-containing diet and were treated with RH1. NQO1 activity in the tumors increased but RH1 produced no antitumor activity in mice fed control or dimethyl fumarate diet. This is consistent with a narrow window of NQO1 activity between no RH1 activation and maximum RH1 activation. This study suggests that selective induction of NQO1 in tumor cells is not likely to be an effective strategy for enhancing the antitumor activity of RH1. In addition, we found that RH1 treatment produced significant leukopenia in mice that may be of concern in the clinic. These results suggest that the ease of reduction of RH1 by NQO1 makes it a poor candidate for an enzyme directed approach to cancer therapy. PMID- 15877231 TI - Light and electron microscopic assessment of progressive atrophy following moderate traumatic brain injury in the rat. AB - The presence of progressive white matter atrophy following traumatic brain injury (TBI) has been reported in humans as well as in animal models. However, a quantitative analysis of progressive alterations in myelinated axons and other cellular responses to trauma has not been conducted. This study examined quantitative differences in myelinated axons from several white and gray matter structures between non-traumatized and traumatized areas at several time points up to 1 year. We hypothesize that axonal numbers decrease over time within the structures analyzed, based on our previous work demonstrating shrinkage of tissue in these vulnerable areas. Intubated, anesthetized male Sprague-Dawley rats were subjected to moderate (1.8-2.2 atm) parasagittal fluid-percussion brain injury, and perfused at various intervals after surgery. Sections from the fimbria, external capsule, thalamus and cerebral cortex from the ipsilateral hemisphere of traumatized and sham-operated animals were prepared and. estimated total numbers of myelinated axons were determined by systematic random sampling. Electron micrographs were obtained for ultrastructural analysis. A significant (P<0.05) reduction in the number of myelinated axons in the traumatized hemisphere compared to control in all structures was observed. In addition, thalamic and cortical axonal counts decreased significantly (P<0.05) over time. Swollen axons and macrophage/microglia infiltration were present as late as 6 months post-TBI in various structures. This study is the first to describe quantitatively chronic axonal changes in vulnerable brains regions after injury. Based on these data, a time-dependent decrease in the number of myelinated axons is seen to occur in vulnerable gray matter regions including the cerebral cortex and thalamus along with distinct morphological changes within white matter tracts after TBI. Although this progressive axonal response to TBI may include Wallerian degeneration, other potential mechanisms underlying this progressive pathological response within the white matter are discussed. PMID- 15877232 TI - Aberrant neuronal-glial differentiation in Taylor-type focal cortical dysplasia (type IIA/B). AB - Focal cortical dysplasia (FCD) type IIA/B (Taylor type) is a malformation of cortical development characterized by laminar disorganization and dysplastic neurons. FCD IIA and FCD IIB denote subtypes in which balloon cells are absent or present, respectively. The etiology of FCD IIA/B is unknown, but previous studies suggest that its pathogenesis may involve aberrant, mixed neuronal-glial differentiation. To investigate whether aberrant differentiation is a consistent phenotype in FCD IIA/B, we studied a panel of neuronal and glial marker antigens in a series of 15 FCD IIB cases, and 2 FCD IIA cases. Double-labeling immunofluorescence and confocal imaging revealed that different combinations of neuronal and glial antigens were co-expressed by individual cells in all cases of FCD IIA/B, but not in control cases of epilepsy due to other causes. Co expression of neuronal and glial markers was most common in balloon cells, but was also observed in dysplastic neurons. The relative expression of neuronal and glial antigens varied over a broad range. Microtubule-associated protein 1B, an immature neuronal marker, was more frequently co-expressed with glial antigens than were mature neuronal markers, such as neuronal nuclear antigen. Our results indicate that aberrant neuronal-glial differentiation is a consistent and robust phenotype in FCD IIA/B, and support the hypothesis that developmental defects of neuronal and glial fate specification play an important role in its pathogenesis. PMID- 15877233 TI - Functional profiling of human atrial and ventricular gene expression. AB - The purpose of our investigation was to identify the transcriptional basis for ultrastructural and functional specialization of human atria and ventricles. Using exploratory microarray analysis (Affymetrix U133A+B), we detected 11,740 transcripts expressed in human heart, representing the most comprehensive report of the human myocardial transcriptome to date. Variation in gene expression between atria and ventricles accounted for the largest differences in this data set, as 3.300 and 2.974 transcripts showed higher expression in atria and ventricles, respectively. Functional classification based on Gene Ontology identified chamber-specific patterns of gene expression and provided molecular insights into the regional specialization of cardiomyocytes, correlating important functional pathways to transcriptional activity: Ventricular myocytes preferentially express genes satisfying contractile and energetic requirements, while atrial myocytes exhibit specific transcriptional activities related to neurohumoral function. In addition, several pro-fibrotic and apoptotic pathways were concentrated in atrial myocardium, substantiating the higher susceptibility of atria to programmed cell death and extracellular matrix remodelling observed in human and experimental animal models of heart failure. Differences in transcriptional profiles of atrial and ventricular myocardium thus provide molecular insights into myocardial cell diversity and distinct region-specific adaptations to physiological and pathophysiological conditions. Moreover, as major functional classes of atrial- and ventricular-specific transcripts were common to human and murine myocardium, an evolutionarily conserved chamber specific expression pattern in mammalian myocardium is suggested. PMID- 15877235 TI - Editorial by Prof. Denis. PMID- 15877234 TI - Effects of aging on pressure-induced MAPK activation in the rat aorta. AB - With increasing age, the cardiovascular system experiences substantial alterations in cellular morphology and function. Whilst the factors regulating these changes are unknown, the mitogen-activated protein kinase (MAPK) pathways have emerged as critical components for mediating numerous cellular responses including control of cell growth, differentiation and adaptation. Here we compare the expression, basal activation and the ability of increased pressure to activate the MAPK pathways in adult (6-month-old), aged (30-month-old) and very aged (36-month-old) Fischer 344xBrown Norway F1 hybrid rats. Histochemical analysis demonstrated an age-related increase in tunica media thickness of approximately 11 and 21% in aortae from aged and very aged animals, respectively. Western blot analysis of the MAPK family extracellular signal-regulated kinase (ERK 1/2), p38, and c-Jun NH2 -terminal kinase (JNK) MAPKs showed differential expression and activation among these proteins with age. Expression of ERK 1/2, p38, and JNK were unchanged, slightly increased (10+/-17.5%) or significantly increased (72.3+/-27%), respectively, in very aged aortae. In contrast, basal activation levels of these proteins were reduced (-26.2+/-7.4%), markedly increased (97.0+/-16.8%), and slightly increased (14.4+/-4.5%), respectively, in very aged compared with 6-month rat aortae. An acute increase of aortic intraluminal pressure (200 mmHg) indicated that ERK 1/2 regulation differed from p38 or JNK. Pressure loading-induced phosphorylation of ERK1/2 was unchanged or increased with aging while p38 and JNK phosphorylation was attenuated (P<0.01). These observations confirm previous conclusions that MAPK proteins are regulated mechanically and expand these studies to suggest that MAPK expression and the control of activation are changed with aging. PMID- 15877236 TI - Glycosaminoglycans in childhood urinary tract infections. AB - It has been suggested that urinary glycosaminoglycans (GAGs) form a natural defense mechanism against urinary tract infections (UTIs). This study investigated whether urinary GAGs play a role in pediatric UTIs, and whether urinary GAG level can be used to differentiate upper UTI from lower UTI. Forty one children with UTIs (33 girls and eight boys; mean age 5.4+/-3.7 years) and 46 age- and sex-matched healthy children (35 girls and 11 boys; mean age 6.6+/-3.9 years) were included in the study. Urinary GAG levels were measured at the onset of acute infection and after a 10-day course of antibiotic treatment. Group GAG findings were compared, and comparisons were also made with the patients divided according to sex and according to UTI type (upper versus lower). The mean urinary GAG level in the patient group at the onset of acute infection (pretreatment) was significantly higher than the mean level in the control group (132.2+/-104.8 mg/g vs 42.2+/-27.1 mg/g creatinine, respectively; P <0.01). In the patient group, the mean urinary GAG level after antimicrobial therapy was significantly lower than the pretreatment level (75.9+/-52.1 mg/g vs 132.2+/-104.8 mg/g creatinine, respectively; P <0.01). However, the mean post-treatment level was still higher than the mean level in the controls ( P <0.05). There was no significant difference in urinary GAG levels when patients were categorized as upper versus lower UTI ( P >0.05). The study results suggest that GAGs play an important role in the pathogenesis of UTIs in children, and that measurement of urinary GAGs may be a valuable noninvasive method for evaluating UTIs in this patient group. However, this assay cannot be used to differentiate upper UTI from lower UTI in children. PMID- 15877237 TI - [Pre-hospital airway management in trauma]. AB - Knowledge of trauma is as old as mankind and survived a long way to reach new technologic developments. Being a multisystemic disease, trauma is an important social and financial problem especially in developing countries. Anesthesists possess important roles in the multisystemic management of trauma patients. In our country prehospital life support for trauma victims is primarily realized and arranged by the emergency help and rescue team 112 which belonged to the Ministry of Health. Several courses are arranged for the team especially about endotracheal intubation and resuscitation in the field. Acute major airway injuries require an early diagnosis and a prompt treatment to improve their outcomes. Typical injuries causing alterations in oxygenation or ventilation are head, spinal cord and thoracic traumas. The use of cuffed tracheal tube remains the definitive 'gold standard' for airway control. Tracheal tube placement requires experience and high degree of skill. The laryngeal mask airway, intubating laryngeal mask airway and oesophageal-tracheal combitube are also indicated in maintaining control of airway during emergency trauma patients in the field. A probable occurrence of cervical spine fracture must be assumed while maintaining airway patency. On-site airway aspiration, and monitoring of carbon dioxide must be realized in trauma victims. PMID- 15877238 TI - The effect of epidermal growth factor on anastomosis, fascia, and skin wound healing. AB - BACKGROUND: The effect of epidermal growth factor (EGF) on gastrojejunostomy (anastomosis), fascia, and skin wound healing in rats was investigated. METHODS: The animals (rats) were separated into two main groups. In the first group (control group n: 23), rats were fed on standard diet and tap water without administration of EGF. In the second group (EGF group, n: 19), EGF was added to the diet. Skin, fascia, and anastomosis wounds were created on three different locations via laparotomy and gastrojejunostomy in all rats. In both groups, the rats divided into three subgroups were sacrificed on the 3rd, 7th, and 21st days post operatively. Tensile strength of skin and fascia and bursting pressure strength were measured for wound healing in both groups on 3rd, 7th, and 21st days. RESULTS: Anastomosis tensile strength of EGF group on the 3rd day of postoperation was found to be significantly higher than that of the control group and when we compared anastomosis bursting pressure, fascia tensile strength, and skin tensile strength in both groups, we did not find any significant differences. CONCLUSION: The measurements indicated that on the 3rd day EGF administered group, which had a more remarkable fibroblastic activity at gastrojejunostomy site, was superior to the control group only in terms of anastomosis breaking tensile strength. PMID- 15877239 TI - The effect of femoral lengthening on skeletal muscle: an experimental study in rats. AB - BACKGROUND: To investigate the muscular changes occurring during femoral lengthening performed in 42 rats. METHODS: The rats were randomly divided into six groups and different rates of lengthening were utilized and the rats were sacrificed at the end of the distraction. Histopathologic and histomorphometric measurements were done for all specimens. The results were analyzed statistically by Mann-Whitney U test. RESULTS: By increasing the rate of lengthening, the severity of pathologic changes such as degeneration of muscle fibers, number of vacuoles and cellular infiltration was increased. Phagocytosis of degenerated muscle fibers was observed after 14 days and cellular infiltration decreased sharply and fibrosis formed in the vacuoles and around the muscle fibers after 21 days of lengthening. Cellular infiltration was lost, the muscle had gained its normal appearance and endomysial, perimysial and epimysial fibrosis formed in the group, which were observed for 31 days. Twenty one percent of muscle fibers were atrophic but the ratio of type I to type II muscle fibers did not change. CONCLUSIONS: The findings of the current study showed that different regions with different phases of degeneration occur in the same muscle and irreversible changes in the muscle may develop even in 10 percent lengthening. PMID- 15877240 TI - [The effects of melatonin and pentoxiphylline on L-arginine induced acute pancreatitis]. AB - BACKGROUND: It has been showed that free oxygen radicals and cytokines contribute to tissue damage and impairment of pancreatic microcirculation in acute pancreatitis. In this study, the effects of melatonin and pentoxiphylline were investigated in rabbits with L-arginine induced acute pancreatitis. METHODS: Rabbits were divided into 5 groups (n=10). Any procedure was not applied for the control group (G1). Acute pancreatitis was induced in one group (G2). Melatonin (G3), pentoxiphylline (G4) and melatonin + pentoxiphylline (G5) were given to other groups after induction of acute pancreatitis. Plasma levels of MDA, amylase, LDH, SGOT, IL-6 and TNF-a were measured at 0, 6, 12, 24 and 48 hours and pancreatic tissue was assessed histopathologically. RESULTS: Melatonin significantly reduced amylase activities at 6, 12, 24, and 48 hours (p<0.025), and all biochemical parameters, (excl. MDA) and edema and necrosis of acinar cells after 48 hours. Although pentoxiphylline reduced abnormally increased parameters in acute pancreatitis (significant for SGOT at 6, 12, 24 and IL-6 at 12, 48 hours), it did not normalized pancreatic abnormalities. CONCLUSION: Melatonin in contrast to pentoxiphylline significantly improved biochemical and histopathological abnormalities due to its powerful antioxidant and free oxygen scavenger properties in acute pancreatitis, and it can be used for patients with pancreatitis. PMID- 15877241 TI - Wrist ligaments: their significance in carpal instability. AB - BACKGROUND: Understanding the exact contribution of the supporting ligaments to the functional integrity of the wrist is crucial for the diagnosis and treatment of carpal instabilities. The present study evaluates functional significance of the wrist ligaments with respect to carpal instabilities. MATERIALS AND METHODS: Sixteen fresh cadaver wrists were dissected. Extrinsic and intrinsic ligaments of the wrists (ligamentum radioscaphocapitatum, ligamentum radiolunotriquetrum and ligamentum triquetrohamatocapitatum) were sectioned sequentially. After sectioning of each ligament, the wrist was examined for clinical signs of instability such as misalignement of carpal bones, limited range of motion and dorsal translation. When instability was suspected, radiographs were taken and if instability was confirmed, then the ligament was repaired. RESULTS: Although none of the dorsal ligaments sectioning resulted in instability, sectioning of ligamentum scaphotrapeziotrapezoideum, ligamentum radioscaphocapitatum, ligamentum radiolunotriquetrum and ligamentum triquetrohamatocapitatum displayed scaphotrapeziotrapezoidal, dorsal intercalated segment, lunotriquetral and capitohamate instability respectively. In two wrists with arthrosis, sectioning of all ligaments didn't lead to any instability. CONCLUSION: Instability of the wrist can be classified on anatomical basis after the name of these four ligaments involved i.e. l. scaphotrapeziotrapezoideum, l. radioscaphocapitatum, l. radiolunotriquetrum and l. triquetrohamatocapitatum respectively . This approach clarifies the etiology and treatment of carpal instabilities. PMID- 15877242 TI - [A critical overview of surgical treatment methods of colorectal injuries]. AB - BACKGROUND: There is still ongoing debate on therapeutic results of traumatic colorectal perforations. The aim of this study was to evaluate and compare the incidence, management, and outcome of patients with traumatic colonic perforations. METHODS: From January 1997 to December 2002, 85 surgical patients underwent emergency operation for traumatic colonic perforations. Patients were reevaluated by analyzing the relationship between the overall morbidity and mortality and the surgical treatment options. We took into consideration the time interval between perforation and treatment as well as the nature, the site, and the cause of perforation, patients' age, and additional organ injuries. RESULTS: Mean age was 32,1. The most injured segment was transverse segment of the colon. Hartmann's procedure was performed in 3 patients (3.5%); resection, and primary anastomosis in 11 patients (12%); primary repair in 57(67%); primary repair and proximal diverting colostomy in 11(%12) ; colostomy in 5(5.8%); and appendectomy in 1 patient. Four postoperative death and one fecal fistula occurred. Deaths were not related to the surgical therapy applied. CONCLUSION: Our early postoperative results after primary repair, and resection anastomosis were good. We conclude that either primary repair or resection and anastomosis can be performed with acceptable morbidity for perforations of the colon and rectum. PMID- 15877243 TI - The efficacy of non-operative management in childhood blunt hepatic trauma. AB - BACKGROUND: In this study the results of non-operative management of pediatric hepatic injury after blunt abdominal trauma were evaluated. METHODS: Multitrauma patients (n = 498) admitted between 1998 and 2002 were analysed as for mechanism of trauma retrospectively. Liver injuries were classified according to the American Association for the Surgery of Trauma's Organ Injury Scaling System. Liver function tests, transfusion status, duration of hospital stay, complications, and operative intervention needed were recorded. RESULTS: Seventy five patients had liver injuries. The mean age was 6.7 years and male/female was 2.5/1. Number of patients in terms of injury grade: I: 15, II: 26, III: 29, IV: 4, V: 1. There was no significant difference between mean blood pressures, heart rates, and hematocrite values on admission and post-stabilisation. In three of 21 transfused patients, it exceeded 40 ml/kg and two of them were operated. The mean pediatric trauma score was +2. As a complication biloma was encountered in one patient. One patient with grade V hepatic injury died in the operating room because of heavy bleeding. The average hospital stay was 8,6 days. CONCLUSION: Our results support the efficacy of non-operative management of any grade hepatic injuries due to blunt abdominal trauma, with resultant low complication and mortality rates and shorter hospital stays. PMID- 15877244 TI - [Relationships among ultrasonographic and demographic, clinical, laboratory findings of patients with acute cholecystitis]. AB - BACKGROUND: To evaluate correlations among ultrasonographic, demographic, clinical and laboratory findings of patients with acute cholecystitis. METHODS: The patients older than 17 years of age with acute colecystitis admitted to the general surgery clinics between January 1991 and December 2000 were evaluated and compared in terms of various parameters. RESULTS: 336 (female, 212; male, 124) patients were included in the study. Mean age was 55.71+/-15.10. Two hundred and seventeen patients presented with more than 12 hours of pain, and 277 patients had multiple biliary stones. Gallbladder wall thickness (GWT) was found to be < 3 mm in 223, and > 5 mm in 58 patients. Pericolic fluid (PCF), distended gallbladder, sonographic Murphy positivity were found in 7.7%, 27.7%, and 9.2% of the cases. PCF was significantly higher in patients who had pain for more than 12 hours. Unlike right upper quadrant tenderness and Murphy sign, localized rebound, rigidity, and percussion tenderness showed significant correlations with abnormal USG findings. Leukocyte levels correlated significantly with PCF, multiple stones, GWT (>5mm) and distended gallbladder. Complications were significantly higher in patients with over 5 mm GWT and PCF. CONCLUSION: Due to significant correlations with abnormal ultrasonographic findings and the abovementioned parameters, prospective studies to evaluate these parameters for the diagnosis of acute cholecystitis are required. PMID- 15877245 TI - [Surgical approach for the septagenerian patients with type A aortic]. AB - BACKGROUND: We analysed early and late results of type A aortic dissection in patients over 70 years of age. METHODS: Sixteen patients over 70 years of age, operated for type A aortic dissections were analysed retrospectively. The mean age of the patients were 72.1+/-1.6 years. Five patients were operated on emergent basis for acute and 11 in elective conditions. Deep hypothermic circulatory support was used in 4 patients. Retrograd cerebroplegia was used in 7 cases. The mean duration of follow-up was 33.2+/-43.5 months. RESULTS: The rate of early mortality was 18.8% (n=3) due to multiorgan failure. In 4 cases, excess drainage from the chest tubes were noted, and one patient required reoperation for bleeding. Two patients had neurological complications. In 2 patients with acute dissections, low cardiac output syndrome developed. Although they didn't have coronary artery disease, hemodynamic improvement was noted with inotropes. One patient with acute aortic dissection had renal failure postoperatively. Two patients, one of whom with a history of chronic obstructive pulmonary disease, had respiratory problems postoperatively. CONCLUSION: High variability in mortality rates indicates that advanced age, solely is not an independent predictor of death. Assuming age as a complementary factor defining patient's clinical status will be helpful for more accurate clinical judgement. PMID- 15877246 TI - [Epidemiologic and clinical features of cases applying to Celal Bayar University emergency unit with head trauma]. AB - BACKGROUND: Head trauma (HT) patients constitute a major part of referrals to emergency unit (EU). We aimed to evaluate cases with HT who applied to our emergency unit retrospectively. METHODS: A hundred and seventy seven HT cases who applied to EU between January 1, 2001-June 30, 2003 were analyzed. Age, gender of the patients, time of intervention, type of trauma, level of consciousness, severity of trauma, concomitant systemic trauma, neuroradiological evaluation, intervention in EU, operation requirement and outcomes were recorded. RESULTS: Patients (men, 73.44% and women 26.55%) referred because of traffic accidents (32 +/- 19 yrs; 59.88%) and other etiologies (21 +/- 17 yrs; 40.11%). According to Glasgow coma scale (GCS) 79.1% had minimal or mild (GCS: 13-15), 3.95% had moderate (GCS: 9-12), and 16.95% had severe HT (GCS: 3-8). On craniography fracture was present in 25.99%, and on cranial CT lesions were present in 25.99% of the cases. Some of them (18.64%) were discharged after first examination, and 14.12% after short observation. They (24.29%) were admitted to neurosurgery, and 4.52% of them to intensive care units; 4.52% of the cases died in EU; 9.04% of the patients underwent neurosurgical operations. CONCLUSION: While traffic accidents constitute the primary cause of adulthood HT, falls take the lead during childhood. HT is usually of minimal or mild severity. Collaboration in patient transport, first aid and EU are important factors affecting prognosis of HT. PMID- 15877247 TI - [Treatment of two-part proximal humeral fractures with external fixators]. AB - BACKGROUND: We aimed to evaluate the results of the closed manipulation or transcutaneous reduction and external fixation in the treatment of two-part fractures. METHODS: Eight patients with two-part proximal humerus fractures who had been treated in our clinic with closed manipulation or transcutaneous reduction and external fixation between 1996-2001 were evaluated for this study. We evaluated the final functional status according to the Neer's classification system. RESULTS: Mean age of the patients was 42 (21-75) years. The major aetiologic factor was motor vehicle accident (in 5 cases). Follow-up time was 2 to 4.5 years. Mean union time was 16 weeks (10 to 24). Nonunion and avascular necrosis of the humeral head have not been observed in none of the cases. Superficial pin tract infection was developed in three cases. All of them were treated successfully with antibiotics. Functional results according to Neer's classification were as follows: 5 good. 2 fair and 1 failed. Mean score 77, 25 (50 to 89). CONCLUSION: External fixation of displaced two-part fractures of proximal humerus can be assessed as a reliable method with respect to satisfactory fracture reduction and stability, low rate of complications and good early functional results-obtained with this technique. PMID- 15877248 TI - [Tracheal rupture during esophagectomy]. AB - Traumatic and iatrogenic injuries of the trachea are rare. The most common causes are motor vehicle, especially motorbike accidents. The incidence of tracheal rupture during esophagectomy is reported as 4-10 %. This complication may occur during the dissection of the upper segment of the esophagus from the posterior membranous trachea, especially when there are adhesions. We reported a tracheal rupture during esophagectomy in a 29 year old man with cancer at the post cricoid region of the trachea, which had been irradiated for three months before the operation. While the esophagus was being dissected from the trachea, a vertical rupture extending to carina occurred on the posterior wall causing severe hypoventilation. To improve ventilation the cuffed armored tubes were inserted into both main bronchi which were connected to a "Y" piece. Based on this case we discussed casualties, risk factors, symptoms and anesthetic management of tracheal rupture. PMID- 15877249 TI - [Chronic duodenal stenosis and periduodenal fibrosis secondary to a intramesenteric cyst arising from an old traumatic hematoma: a case report]. AB - A case of mesenteric fibrosis secondary to post-traumatic mesenteric hematoma, resulting in duodenal stenosis eight years after the traumatic event is presented. A 50 year-old man was hospitalized with nausea and vomiting. An intraabdominal cystic mass was seen in abdominal ultrasonography and CT. Duodenum was found to be obstructed due to extrinsic compression in upper endoscopy. At laparotomy there was an intramesenteric traumatic cyst and fibrosis around the duodenum resulting in duodenal obstruction. After the cyst was unroofed a Roux-en Y duodenojejunostomy was performed. Both early and late (after 4 months) follow up results were excellent. PMID- 15877250 TI - Duodenal intramural hematoma due to blunt abdominal trauma. AB - A 14-year-old boy was admitted to the hospital with the complaints of epigastric pain and vomiting for three days following epigastric trauma. Physical examination revealed a palpable mass in the epigastrium without any signs of peritoneal irritation. Vital signs, biochemical analysis, chest and abdominal x rays were normal. Abdominal US, CT and upper gastrointestinal endoscopy showed that there was a well-defined duodenal intramural mass suggesting hematoma. Intraabdominal solid organs were normal and we did not observe free air or extravasation of contrast media. Therefore, we followed up the patient by US. The hematoma completely resolved 48 days after the trauma. PMID- 15877251 TI - [Incarceration of the left hepatic lobe in incisional hernia: a case report]. AB - Incisional hernias are delayed complications of abdominal surgery and occur in 0.5-13.9 % of patients according to various reported series. The causative factors of incisional hernia are related to the condition of the patient, the main disease, surgical techniques and postoperative complications. The subcutaneous herniation of the left hepatic lobe through the abdominal wall is a rare condition. In this case report, we discuss an incarcerated incisional hernia associated with medial segment of the left hepatic lobe. PMID- 15877252 TI - Adjunctive hyperbaric oxygen therapy contributes healing in electrical injury: a case report of high voltage electrical injury. AB - In electrical injuries, new treatment modalities and guidelines are needed for improving clinical outcome and the survival of damaged tissue. Although there is no published study about hyperbaric oxygen (HBO) therapy for electrical injury in the literature, it is indicated in conditions, which may contribute to the clinical presentation of electrical injury such as thermal burns, crush injuries, necrotizing soft tissue infections, problematic wounds and compromised skin grafts and flaps. An 11-year-old child with high voltage electrical injury treated with adjunctive hyperbaric oxygen for 90 minutes twice a day at 2,4 ATA for one week, then once a day for six days for a total of 20 sessions was presented to demonstrate the beneficial effects of hyperbaric oxygen therapy initiated before irreversible damage had taken place. Although hyperbaric oxygen therapy was initiated rather late, when the most effective window for intervention had already past, HBO was effective in fighting against necrosis, infection and tissue loss. Adjunctive HBO therapy is suggested for electrical injuries for its contribution to healing. In order to see the favourable effects of HBO, it is better to start the treatment within the first 24 hours following injury. PMID- 15877253 TI - A false aneurysm mistaken for a DVT after hip surgery. AB - False or pseudoaneurysm formation usually occurs after traumatic, iatrogenic or infective injury to the arterial wall. Despite the high incidence of pseudoaneurysm formation secondary to puncture injury to the common femoral artery false aneurysm formation of the profunda femoris artery (PFA) is a rare complication and has not been previously reported as a complication of orthopaedic surgery. We present a patient who developed a false aneurysm of the PFA secondary to arterial damage caused by a bone fragment dislodged during orthopaedic surgery for fracture of the femur. PMID- 15877254 TI - Delay of gastric emptying measured by 13C-acetate breath test in neurologically impaired children with gastroesophageal reflux. AB - Delayed gastric emptying often occurs in patients, including children, with gastroesophageal reflux (GER) due to neurological impairment (NI). The aim of this study was to evaluate gastric emptying of liquids in children with symptomatic GER using the (13)C- acetate breath test (ABT), and to compare the gastric emptying rates between children without and with NI. Thirteen patients were divided into 2 groups: group I without NI (5 patients) and group II with NI (8 patients). The liquid test meal consisted of Racoltrade mark (5 ml/kg) mixed with (13)C- acetate (50 mg for infants, 100 mg for children, and 150 mg for adolescents). Breath samples were collected for (13)CO (2) measurement before the intake of the meal, every 15 minutes during the first 2 hours after the meal and every 30 minutes thereafter to assess the ingestion of (13)C- acetate and Racoltrade mark. (13)CO (2) was measured using a gas chromatograph-isotope ratio mass spectrometer. The results were expressed as % of (13)C expired per hour and cumulative (13)C excretion over a 3-hour period. The half excretion time in (13)C ABT(t (1/2) ex) was 1.095 hour for group I and 1.817 hour for group II (p = 0.0045). The lag time (t lag), which reflected the initial delay of gastric emptying, was 0.666 h for group I and 1.002 h for group II (p = 0.0045). Gastric emptying studies can be easily and reliably carried out in children with NI using (13)C- ABT. We showed that we were able to determine the gastric emptying rate by (13)C- ABT in patients with GER due to NI. PMID- 15877255 TI - Extrahepatic portal hypertension: quality of life and somatic growth after surgery. AB - AIM: This study was carried out in order to determine whether children with extrahepatic portal hypertension show any improvement in growth parameters and quality of life after elective surgery. PATIENTS AND METHODS: All patients with extrahepatic portal hypertension who underwent surgery between April 1999 and March 2002 were studied prospectively. Height and weight before and after surgery were converted into Z scores for comparison. The quality of life was assessed by changes in scholastic ability, physical activity, social interaction, and economic effects on the family. RESULTS: At the end of a minimum follow-up period of 1 year, of the 30 children, 50 % and 76 % showed improvement in weight and height Z scores, respectively, compared to their scores while on medical management. Z scores below - 2 SD for weight and height were seen in 57 % and 37 % of patients prior to surgery, and only in 40 % and 20 %, respectively at the end of follow-up, which ranged from 1 - 4 years. The improvement in height and weight was more in those who had undergone splenectomy with either devascularization or central splenorenal shunt compared to those who had undergone side-to-side lienorenal shunt without splenectomy. The children, who had dropped out of school while on medical management, either rejoined school or attended vocational courses after surgery. There was an improvement in school performance and physical activity in 85 % of the children. Improvement in personality was seen in 73 % of the affected patients. The cost incurred for surgery was one-third of the amount spent while on medical management. CONCLUSIONS: Growth parameters improved significantly in children after surgical intervention for portal hypertension. Overall improvement in scholastic abilities, physical activity, and social interaction was noted in a majority of the patients. As a one-time procedure in a developing country, surgery is also more cost effective. PMID- 15877256 TI - Comparative study between isolated intestinal perforation and necrotizing enterocolitis. AB - INTRODUCTION: Intestinal perforations in the neonatal period are usually related to necrotizing enterocolitis (NEC) or intestinal occlusion. Intestinal perforation in the absence of these conditions is called isolated perforation (IP). Several risk factors and pathogenic mechanisms have been suggested, and most of them are common to those classically attributed to NEC. AIM: To identify and compare the clinical and pathological features of IP and NEC. MATERIAL AND METHODS: We reviewed all cases of neonatal intestinal perforation and NEC in the last five years. Thirty-three patients were retrospectively classified into Group NEC: 24 cases, and Group IP: 9 cases. We collected multiple data as study variables: 1) General features; 2) Obstetric history; 3) Neonatal treatment; 4) Comorbidity; 5) Perforation features; 6) Treatment and outcome. RESULTS: Comparing the groups, we found statistical significant differences in isolated perforation cases with these risk factors: extreme prematurity, very low birth weight, abruptio placenta, intubation and neonatal mechanical ventilation, umbilical catheterization, precocious sepsis, and indomethacin therapy. A more precocious operation and a good prognosis also reached statistical significance. In the other hand, we found statistically significant differences in NEC with congenital cardiopathy (excluding isolated patent ductus arteriosus), with intestinal pneumatosis, with diffuse bowel involvement and a worse prognosis. Risk factors and pathologic findings seem to support an ischaemic pathogenesis in both diseases. PMID- 15877257 TI - Outcome and long-term growth after extensive small bowel resection in the neonatal period: a survey of 87 children. AB - This retrospective study aims to analyze the outcome, the prognosis factors and the long-term growth of children after extensive small bowel (SB) resection in the neonatal period. PATIENTS AND METHODS: 87 children, born between 1975 and 1991 who had undergone extensive neonatal small bowel resection, were followed up over a mean period of 15 years. Anatomical data influencing PN dependency and duration were analyzed. Data on height and weight were collected and compared using growth standards. Final heights were studied for patients who achieved their puberty and compared to predicted height based on Tanner's formula. Patients were analyzed according to PN weaning and growth: children still receiving PN (group A), patients weaned from initial PN but requiring PN once again or enteral feeding (group B), and children with permanent intestinal autonomy (group C). RESULTS: The overall survival is 89.7 %, depending on the date of birth. The duration of PN-dependency varies according to the intestinal length and the presence of the ileocaecal valve (ICV). All patients who remain PN dependent had less than 40 cm of small bowel and/or the absence of ICV. Patients in group B had a mean small bowel length of 35 +/- 19 cm, resection of the ICV in 50 % of cases, and a PN duration of 47.4 +/- 23.8 months. There was a significant decrease in height and weight gain within the 4 years after cessation of PN, requiring enteral or parenteral feeding. Patients in group C had a mean small bowel length of 57 +/- 19 cm, presence of ICV in 81 % of cases and a PN duration of 16.1 +/- 11.4 months. After PN weaning, they grow up normally with normal puberty and final height as predicted from genetic target height. CONCLUSION: PN duration is influenced by the length of residual SB and the absence of ICV. With good anatomic prognosis factors and short duration of initial PN, normal long term growth may be predicted. Conversely, poor anatomical factors and protracted initial PN require careful monitoring of growth and may sometimes require nutritional support to be restarted. The last group, permanently dependent on PN, might be candidates for intestinal transplantation. PMID- 15877258 TI - The histopathology of congenital pouch colon associated with anorectal agenesis. AB - The colonic pouch excised during surgery in 17 patients with congenital pouch colon associated with anorectal agenesis (CPC) was subjected to detailed histological examination after staining with hematoxylin and eosin. The patients included 11 newborns, 3 infants, and 3 older children. The most frequently observed abnormalities were acute and chronic inflammation of the mucosa and submucosa, focal or generalized thinning of muscle layers, especially of the outer muscle coat, disorganized muscle layers, a decreased number of mature ganglion cells, and neuronal hyperplasia and hypertrophy in nerve plexuses. Ectopic heteroplastic tissues were identified in 2 patients. These findings suggest that the colonic pouch in CPC represents abnormally developed colonic tissue and points to the similarity with segmental dilatation of the colon. The neuromuscular abnormalities explain the physiological characteristics of the colonic pouch, namely weak peristalsis as well as the propensity to undergo marked dilatation even after tubularization. PMID- 15877259 TI - Effects of pentoxifylline and vitamin E on the bilateral ovary after experimental ovarian ischemia. AB - Pentoxifylline (PTX) and vitamin E inhibit the release of superoxide and hydroxyl radicals, and PTX improves capillary flow and tissue oxygenation. This experimental study was designed to determine the effects of PTX and vitamin E in the ovary after unilateral ovarian ischemia reperfusion (I-R) in albino Wistar rats. A vascular clamp was placed on the left ovary for 4 hours in all groups except for the control group. Following this, in the ischemia (I) group bilateral ovariectomy was performed. Saline, PTX, vitamin E, and PTX plus vitamin E were infused 30 min before reperfusion in the reperfusion (R), pentoxifylline (P), vitamin E (E), and pentoxifylline plus vitamin E (PE) groups, respectively. After 4 hours of reperfusion, the ovaries were removed for biochemical and histologic examination. MDA levels of bilateral ovaries in the PE group were significantly lower than in the E group (p < 0.0033). NO levels of bilateral ovaries in the PE group were significantly lower than in the P and E groups (p < 0.0033). Massive hemorrhage was determined in the ipsilateral ovaries of the R group. Hemorrhage was minimal or moderate in the ipsilateral ovaries of other groups. The contralateral ovaries showed congestion in different degrees. The contralateral ovaries of the group PE and the bilateral ovaries of the control group showed no pathological changes. PTX and vitamin E given together seems to be more effective in reducing I-R injury in ovarian tissue compared to administration of PTX, or vitamin E alone. However, further studies are required to evaluate the effective dose and duration of PTX and vitamin E on bilateral ovaries. PMID- 15877260 TI - Resveratrol reduces ischemia reperfusion injury after experimental testicular torsion. AB - The aim of the study was to evaluate the effects of resveratrol on testicular ischemia reperfusion injury. Forty Wistar albino rats were divided into 4 groups. Torsions (ischemia) were created by rotating the right testis 720 degrees in a clockwise direction for 4 hours in all groups except the control group. In the torsion group after 4 hours' ischemia bilateral orchiectomy was performed. In the detorsion group, saline was injected by an intraperitoneal route, 30 min before detorsion (reperfusion). In the resveratrol group, 30 mg/kg resveratrol was injected by an intraperitoneal route, 30 min before detorsion. In the detorsion and resveratrol groups, the bilateral testes were removed after 20 hours of detorsion. In all groups, the tissue levels of malondialdehyde (MDA) and glutathione (GSH) and histological changes were determined. In rats treated with resveratrol, MDA levels (138 +/- 25 nmol/mg protein) were significantly decreased compared with torsion (426 +/- 178 nmol/mg protein) and detorsion (370 +/- 76 nmol/mg protein) groups (p < 0.05). GSH levels (6.54 +/- 0.8 micromol/g wet tissue) were significantly increased compared with torsion (4.61 +/- 0.4 micromol/g wet tissue) and detorsion groups (5.24 +/- 0.9 micromol/g wet tissue) (p < 0.05). The mean testicular tissue injury score in the resveratrol group was significantly lower than in torsion and detorsion groups (p < 0.05). The present study demonstrates that intraperitoneal administration of resveratrol in rats may protect testis against injury associated with reperfusion. PMID- 15877261 TI - Purpura fulminans -- soft tissue damage as a manifestation of bacterial sepsis. AB - We present here two cases of purpura fulminans (PF), a rare complication of septicemia. Its fulminant course usually prompt the surgeon into quick and aggressive action. Although sometimes it may lead to mutilation of a patient, nevertheless it is a life-saving procedure. A decision to amputate all four extremities is very difficult, both for the parents and for the surgeon. Therefore one must be absolutely sure that this management strategy is the only way to save the patient's life. Two cases of meningococcal and staphylococcal sepsis treatment are described in this paper. The surgical approach is discussed together with a review of the literature. PMID- 15877262 TI - Multiple and recurrent intestinal perforations: an unusual complication of Henoch Schonlein purpura. AB - Intestinal perforation, intussusception, and infarction constitute the major surgical complications in Henoch-Schonlein purpura. Early corticosteroid treatment for intestinal complications is recommended. Here the authors describe the case of a 13-year-old boy with Henoch-Schonlein purpura who had multiple and recurrent perforations that occurred under corticosteroid treatment. PMID- 15877263 TI - Appendix as a biliary conduit for choledochal cysts in children. AB - Biliary conduits are constructed in operations for choledochal cysts. A wide variety of options are available for biliary tract reconstruction. We present our experience of treating six children with choledochal cysts by using the appendix as a biliary conduit. After mobilizing the appendix on its vascular pedicle, nonrefluxing, tunneled anastomosis was made with the 2nd part of the duodenum and the appendix. The operative procedure was simple and less time-consuming. Postoperative evaluation was done with the help of HIDA scan and ultrasound examination. Postoperative cholangitis was conspicuously absent in the two years of follow-up. One child has been lost to follow-up. From our preliminary experience, the operation seems simple and satisfying. Though the long-term efficacy still remains to be proven, the appendix should prove durable as a functional conduit. PMID- 15877264 TI - One early and three delayed distal pancreatectomies without splenectomy and with preservation of the splenic vessels after traumatic transection of the distal pancreas in children. AB - Four children are presented with Class II pancreas injury as a result of a motor vehicle accident. The first child was taken to the operating room promptly due to concomitant perforation of the hollow viscus (gastric rupture) and underwent successful spleen-sparing distal pancreatectomy with preservation of the splenic artery and vein. The next three cases with isolated abdominal symptoms of pancreatic injury generally experienced a delay of one day before the onset of abdominal symptoms and positive diagnostic investigation results and were managed non-operatively (NOM) on admission; they were then treated surgically due to developing peritonitis after 24, 36, and 38 hours, respectively. The same type of operation, even though delayed and technically much more demanding, was performed, but this was not an obstacle to our efforts to preserve the spleen and its full circulation, in order to provide full immunological and haematological support during the expected prolonged postoperative course. We did not lose any of the children. A reintervention was needed in two children due to the retention of necrotic tissue and intraabdominal abscess. The serum amylase level in all cases remained above normal. We believe that a spleen-preserving distal pancreatectomy with splenic vessel conservation can be safely performed, even in delayed operations, and should be indicated for the surgical management. PMID- 15877265 TI - Entero-enteric fistula following mild necrotizing enterocolitis. AB - We report a 26-week gestational age newborn who developed an entero-enteric fistula associated with a small bowel stricture following a mild episode of necrotizing enterocolitis. The fistula led to a delay in the diagnosis and surgical treatment of the stricture, and should therefore be considered in the setting of partial or intermittent obstruction in the newborn who has had necrotizing enterocolitis. PMID- 15877266 TI - Dorsal trans-scaphoid perilunate dislocation in a 9-year-old child. AB - The authors reported on the surgical management of a trans-scaphoid retrolunate dislocation of the carpus associated with a stable styloid fracture of the radius and a displaced fracture of the triquetrum in a nine-year-old child. An open reduction of the dislocation and fixation using a screw inserted disto-proximally in the scaphoid were quickly carried out using a palmar approach. X-ray showed a good union nine months later. At 29 months the motion of the wrist was normal and pain-free, the strength was graded at 80 % in comparison to the contralateral side and the X-ray examination showed no carpal instability nor growth trouble. The authors discuss the mechanism of production and the therapeutic strategy. PMID- 15877267 TI - [Fractures of the scaphoid tubercle]. AB - BACKGROUND: Avulsion fractures of the scaphoid tubercle (Herbert type A1) are rare. Recommendations in the literature focus on a four to six week plaster immobilization including the thumb. Surgery is recommended in case of additional injuries to the carpus. MATERIAL AND METHODS: In a retrospective investigation including a 36 month period we reviewed acute fractures of the scaphoid and avulsion fractures of the scaphoid tubercle. RESULTS: We found 81 acute scaphoid fractures, four of them (4.9 %) were isolated tuberculum avulsions. Three cases were extra-, one case an intraarticular fracture. The first were treated conservatively, the last with a percutaneous screw. CONCLUSIONS: Following our own experiences with those rare injuries we recommend a differentiation of the fracture type into extraarticular injuries where conservative treatment should still be performed and intraarticular fractures where percutaneous screw osteosynthesis should be discussed. PMID- 15877268 TI - [Wrist lesions in MRI arthrography compared with wrist arthroscopy]. AB - BACKGROUND: Wrist arthroscopy is routinely used for diagnosis of ligamentous lesions to the wrist. Although it is very sensitive and specific, it is also more invasive than other available diagnostic techniques. METHOD: In a prospective trial, 125 patients (80 men, 45 women) 37 years old (+/- 12) with clinical evidence of wrist lesions were evaluated with direct wrist arthro MRI. Within 24 hours following direct arthro MRI wrist arthroscopy was performed. The surgeons and radiologists were not aware of the other results until completion of their investigation. Specificity, sensitivity, positive predictive value, negative predictive value and accuracy were determined. RESULTS: According to the MRI findings, lesions of the TFCC were suggested in 70 patients (56 % of all patients). In 65 cases this was confirmed by wrist arthroscopy. In the remaining 55 patients no TFCC lesions were detected by MRI. However in three cases TFCC lesions were found by arthroscopy. In the remaining 52 patients MRI accurately excluded TFCC lesions. There was a correlation of MRI and arthroscopy in detecting TFCC lesions in 93.6 %. Sensitivity was 94 %, specificity 89 %, positive predictive value 91 % and negative predictive value 93 %. Complete scapholunate ligament tears were detected by MRI in 12 (9.6 %) cases. Correlation with wrist arthroscopy was 99 %, sensitivity 92 %, specificity 100 %, positive and negative predictive value 100 % and 99 %, respectively. Accuracy for incomplete scapholunate lesions (n = 17, 13.6 %) and lunotriquetral tears (n = 4, 3.2 %) was poor (sensitivity 59 % and 25 %, specificity 100 % and 99 %). CONCLUSION: Though sensitivity of MRI arthrography approaches that of arthroscopy, it cannot replace it at the moment. However, it is a potent additional tool for wrist diagnosis if intraarticular contrast is used. It can facilitate diagnosis and indications for surgery of the wrist. It may make arthroscopic and more invasive interventions for diagnostic purposes avoidable in future. PMID- 15877269 TI - [Results of ligament reconstruction in advanced scapholunate dissociation]. AB - Scapholunate dissociation is one of the most common disorders of the wrist. Untreated it might lead to osteoarthrosis (scapholunate advanced collapse, SLAC wrist). Choosing the best surgical treatment option is still challenging, especially in cases of carpal collapse in combination with beginning osteoarthrosis of the radial styloid and the proximal pole of the scaphoid. We report the results of a homogenous group of eight patients with reducible carpal collapse and beginning arthrosis treated by reconstruction of the scapholunate ligament. The operation was performed 66 (range: 20 to 252) months after trauma. The average length of follow-up was two years. Five patients stated general improvement, while three reported a change for the worse. At follow-up, the average total range of motion of the operated wrist was decreased by 16 % compared to the unaffected side. The average grip-strength (measured with a Jamar dynamometer) was 77 % of the uninvolved wrist. The DASH score was 43 +/- 25. In three cases the Martini score showed a good or an excellent result. The average scapholunate angle was 72.3 degrees preoperatively and decreased to 61.0 degrees at follow-up. At follow-up as well as pre- and postoperatively the carpal height ratio showed pathologic mean values. Therefore, reconstruction of the carpal alignment was not achieved in most of the cases. Progression of the osteoarthrosis has to be expected. Reconstruction of the scapholunate ligament for treatment of carpal collapse with beginning osteoarthrosis therefore remains an unsolved problem. PMID- 15877270 TI - [Dorsal capsulodesis versus triscaphe arthrodesis in patients with scapholunate dissociation. Do the results depend on the diagnosis or the operation?]. AB - BACKGROUND: Dorsal capsulodesis and triscaphe arthrodesis are possible treatment modalities for patients with scapholunate dissociation. In light of overlapping indications for either operation for patients with carpal instability, it is important to compare the postoperative results. METHODS: From 1998 until 2002, we operated on 87 patients suffering from scapholunate dissociation. 52 patients were treated by dorsal capsulodesis after R. Berger and 35 patients were treated by triscaphe arthrodesis as published by Watson. Of these 87 patients, we managed to follow-up 77 patients (47 dorsal capsulodesis und 30 triscaphe arthrodesis) with a physical examination, X-ray of the wrists and Krimmer Score. RESULTS: In terms of grip-strength, range of motion, functional outcome (Krimmer Score), duration of the operation and hospitalisation, the dorsal capsulodesis group performed better (p < 0.05) than the triscaphe arthrodesis group at the time of follow-up. At follow-up, pain reduction was significant in both groups (p < 0.05). Krimmer Score (functional outcome) and the rate of complication was clearly better for the dorsal capsulodesis group as compared to the triscaphe arthrodesis group. CONCLUSION: In case of non-static scapholunate dissociation, dorsal capsulodesis should be the first choice treatment. In case of a young manually working man with static scapholunate dissociation, it should be a case to-case decision whether performing a triscaphe arthrodesis or a dorsal capsulodesis. With this investigation we wanted to discuss and demonstrate the difficulties with the differential-indication for the two operations. Generally speaking the postoperative results did not depend on the type of scapholunate dissociation (dynamic versus static) but rather on the chosen surgical procedure. PMID- 15877271 TI - [Functional results after proximal row carpectomy (PRC) in patients with SNAC /SLAC-wrist stage II]. AB - The proximal row carpectomy (PRC) is a motion preserving procedure which creates a new joint without arthrosis. It is a frequently used procedure in stage II of a posttraumatic degenerative arthrosis of the wrist after scaphoid nonunion or scapholunate ligament instability (SNAC-/SLAC-wrist). In this retrospective analysis the functional postoperative results of this operation are compared in light of a homogenous indication (SNAC-/SLAC-wrist stage II). In 38 patients PRC was performed for a stage II SNAC- (n = 29) or SLAC-wrist (n = 9) between June 1994 and March 2002. Postoperative examination included range of motion and grip strength. Pain was assessed using a visual analogue scale (VAS 0 - 100). The DASH questionnaire (disability of the arm, shoulder and hand) was used to evaluate the disabilities in activities of daily living (ADL). Thirty patients (79 %) with a mean age of 39 years (23 - 59) were evaluated with a mean follow-up of 27 months (6 - 100). Mean extension and flexion of the wrist reached 75 degree which was 57 % of the contralateral hand. Mean radial and ulnar deviation was 33 degree corresponding with 52 % of the contralateral hand. The average grip strength was 50 % of the unaffected side. The postoperative DASH score was 27.4. Pain with strenuous activity was reduced by 40 %, resting pain by 77 %. Three patients showed radiological signs of a radiocapitate arthrosis, one patient needed conversion into a complete wrist arthrodesis. Our results are in concordance with the literature. However, our follow-up time is relatively short and we cannot make any conclusion about the long-term outcome. PRC is a technically straightforward procedure for treatment of carpal collapse. For stage II of the SNAC-/SLAC-wrist we consider the resection of the proximal carpal row an alternative procedure to the midcarpal arthrodesis particularly in patients who require less grip strength and when a shorter postoperative immobilization is reasonable. PMID- 15877272 TI - [Mid-term results after proximal row carpectomy and review of the literature]. AB - PURPOSE: The aim of this study is to evaluate the results after proximal row carpectomy and to compare them with results in the literature. METHOD: Between 1994 and 2001, 37 patients underwent proximal row carpectomy. 30 patients were available for follow-up. In all cases the proximal row carpectomy was performed through a dorsal approach. Clinical parameters were evaluated by using the conventional wrist score (Krimmer score) as well as the DASH-score. RESULTS: Follow-up examination shows a range of motion (ROM) for wrist extension and flexion of 46 % of the contralateral side. ROM for ulnar and radial deviation is 42 % of the other side, ROM for pronation and supination is equal to the other side. Mean grip strength is determined to be 58 % of the contralateral side. 90 % of the patients are satisfied with the result of the operation. The Krimmer score amounts to 58 and the DASH score to 39 points. CONCLUSION: We consider proximal row carpectomy to be a good therapeutic option for lunate necrosis stage IIIB or IV or carpal collapse stage II (SNAC or SLAC wrist) with a concomitant lesion of extrinsic ligaments with ulnar translocation. Another indication is the acute, non-reconstructable or the chronic perilunar luxation with arthrosis. PMID- 15877273 TI - [Functional outcome after midcarpal arthrodesis in the treatment of advanced carpal collapse (SNAC-/SLAC-wrist)]. AB - PURPOSE: The purpose of this retrospective study was the evaluation of the functional outcome of midcarpal arthrodesis, its results in the treatment of SNAC/SLAC stage II and III and patients' satisfaction. METHODS: 49 patients that were treated with a midcarpal arthrodesis were re-examined at a mean follow-up time of 47 months. Range of motion was verified and grip strength was measured with a Jamar-Dynamometer II and a pinch-grip. Pain was evaluated by a visual analogue scale (VAS 0 to 100) for stress and under resting conditions. Patients' daily activities and general quality of life were estimated with the DASH questionnaire. Radiographic evaluation was done by conventional X-ray. RESULTS: Active range of motion was 56 % and grip strength was 76 % of the non-operated wrist. The DASH score was 29 points. Pain relief was 34 % during resting conditions and 31 % after stress respectively. 45 patients demonstrated bony consolidation in X-ray control. Six patients needed further treatment with a total arthrodesis because of pain or absence of bony consolidation. 77 % of the patients returned to their original occupation and 80 % were satisfied with the final result. CONCLUSION: Our data demonstrate that midcarpal fusion is a reliable procedure for treating the difficult condition of advanced carpal collapse if proper realignment of the carpus is performed. PMID- 15877274 TI - [Does the disability of shoulder, arm and hand questionnaire (DASH) replace grip strength and range of motion in outcome-evaluation?]. AB - Scoring systems currently used to evaluate functional disabilities in upper extremity conditions frequently combine so-called "objective" parameters such as grip strength and range of motion with "subjective" parameters such as pain. Since its introduction, the Disabilities of Arm, Shoulder and Hand Questionnaire has proven useful in the pre- and postoperative evaluation of functional limitations of the upper extremity. This study examines the relationship between DASH scores, the so-called "objective" parameters of grip strength and range of motion and those of "mixed scores" such as Krimmer and Cooney Scores. Based on the data of six diagnostic groups, Spearman's correlation coefficients were calculated. There was no correlation between DASH scores and range of motion, however, some of the diagnostic groups showed a moderate correlation between DASH scores and grip strength. Both the Krimmer and Cooney scores showed a significant correlation with the DASH scores, indicating that these "mixed-scores" can be replaced by the DASH. Since it has been shown that grip strength and range of motion do not capture patients' functional limitations adequately, their continued use in the evaluation of upper extremity conditions should be the topic of further discussion. PMID- 15877275 TI - [Diagnosis of lunotriquetral instability]. AB - BACKGROUND: Lunotriquetral dissociation with rupture of the lunotriquetral ligament and the radiolunotriquetral and the radiodorsal ligament is usually recognized late. Cinematography, arthrography, magnetic resonance imaging and arthroscopy are the diagnostic procedures. Treatment includes nonoperative methods in the case of incomplete lesions but also closed reposition with temporary LT arthrodesis and open ligament repair. In many cases, definitive LT arthrodesis may become necessary. It is the aim to develop a diagnostic algorithm on the base of retrospective analysed data. MATERIAL AND METHODS: From January 1998 to July 2003, 97 cinematographies of the wrist were performed, 22 with the question for a dynamic or static VISI deformity as a sign for lunotriquetral instability. This group of patients was analysed retrospectively to evaluate the diagnostic reliability. Based on these datas, a diagnostical algorithm was established for prospective investigations. RESULTS: The combination of cinematography, arthrography and magnetic resonance imaging suspected lunotriquetral instability in ten cases so that an arthroscopy was arranged. This way LT-lesions were verified in nine cases. The mean interval between accident and first contact with our institution was five months. CONCLUSIONS: The management of lunotriquetral injuries is complicated by the long time from trauma to definitive diagnosis. A combination of refined clinical and apparative investigations can lead to a reliable diagnosis. PMID- 15877276 TI - [Resection arthroplasty of the carpometacarpal joint of the thumb - results of 132 cases]. AB - BACKGROUND: Resection arthroplasty of the carpometacarpal joint of the thumb is considered to be the most frequently used surgical treatment for arthrosis of the trapeziometacarpal joint. To avoid proximalisation and to improve stability of the first metacarpal, tendon-sling resectional arthroplasties have gained general approvement. To further simplify surgery, we have used a surgical technique since 1995, that consists in a fixation of the first metacarpal with local capsular tissue. METHOD AND CLINICAL MATERIAL: From 2000 to 2001, 152 resections of the trapezium were performed in 147 patients with arthrosis of the first carpometacarpal joint. 48 surgical procedures included a tendon-sling arthroplasty; in 104 cases stabilisations of the first metacarpal were achieved by fixing local radiopalmar capsular tissue to the flexor-carpi-radialis tendon. After a mean follow-up of 30 months (18 to 41 months), patients were asked to complete a questionnaire concerning pain, physical strength, practical skills, maximal physical capacity and aesthetic result. The general surgical result had to be scored and the recovery time until the thumb could be used for activities of daily living had to be recorded. RESULTS: 132 of 152 questionnaires, 43 of the patients with tendon-sling arthroplasty and 89 of the patients with arthroplasty with local radiopalmar capsular tissue, were returned. After tendon-sling arthroplasty, 69.8 % of the patients judged their results to be good or very good, after stabilisation with radiopalmar capsular tissue 69.7 % of the patients valued their surgical results good or very good. Unsatisfactory results were found in 18.6 % of the patients after tendon-sling arthroplasty and in 19.1 % of the patients after stabilisation with radiopalmar capsular tissue. The thumb could be used for every-day life after a mean time of 6.7 months. There was no significant difference between the operative procedures. CONCLUSION: The collected data implies that resection-arthroplasty of the carpometacarpal joint of the thumb with stabilisation by radiopalmar capsular tissue yields similar results compared to tendon-sling arthroplasty. PMID- 15877277 TI - Single-gene disorders: what role could moonlighting enzymes play? AB - Single-gene disorders with "simple" Mendelian inheritance do not always imply that there will be an easy prediction of the phenotype from the genotype, which has been shown for a number of metabolic disorders. We propose that moonlighting enzymes (i.e., metabolic enzymes with additional functional activities) could contribute to the complexity of such disorders. The lack of knowledge about the additional functional activities of proteins could result in a lack of correlation between genotype and phenotype. In this review, we highlight some notable and recent examples of moonlighting enzymes and their possible contributions to human disease. Because knowledge and cataloging of the moonlighting activities of proteins are essential for the study of cellular function and human physiology, we also review recently reported and recommended methods for the discovery of moonlighting activities. PMID- 15877278 TI - Joint modeling of linkage and association: identifying SNPs responsible for a linkage signal. AB - Once genetic linkage has been identified for a complex disease, the next step is often association analysis, in which single-nucleotide polymorphisms (SNPs) within the linkage region are genotyped and tested for association with the disease. If a SNP shows evidence of association, it is useful to know whether the linkage result can be explained, in part or in full, by the candidate SNP. We propose a novel approach that quantifies the degree of linkage disequilibrium (LD) between the candidate SNP and the putative disease locus through joint modeling of linkage and association. We describe a simple likelihood of the marker data conditional on the trait data for a sample of affected sib pairs, with disease penetrances and disease-SNP haplotype frequencies as parameters. We estimate model parameters by maximum likelihood and propose two likelihood-ratio tests to characterize the relationship of the candidate SNP and the disease locus. The first test assesses whether the candidate SNP and the disease locus are in linkage equilibrium so that the SNP plays no causal role in the linkage signal. The second test assesses whether the candidate SNP and the disease locus are in complete LD so that the SNP or a marker in complete LD with it may account fully for the linkage signal. Our method also yields a genetic model that includes parameter estimates for disease-SNP haplotype frequencies and the degree of disease-SNP LD. Our method provides a new tool for detecting linkage and association and can be extended to study designs that include unaffected family members. PMID- 15877279 TI - Fatal congenital heart glycogenosis caused by a recurrent activating R531Q mutation in the gamma 2-subunit of AMP-activated protein kinase (PRKAG2), not by phosphorylase kinase deficiency. AB - Fatal congenital nonlysosomal cardiac glycogenosis has been attributed to a subtype of phosphorylase kinase deficiency, but the underlying genes and mutations have not been identified. Analyzing four sporadic, unrelated patients, we found no mutations either in the eight genes encoding phosphorylase kinase subunits or in the two genes encoding the muscle and brain isoforms of glycogen phosphorylase. However, in three of five patients, we identified identical heterozygous R531Q missense mutations of the PRKAG2 gene, which encodes the gamma 2-subunit of AMP-activated protein kinase, a key regulator of energy balance. Biochemical characterization of the recombinant R531Q mutant protein showed >100 fold reduction of binding affinities for the regulatory nucleotides AMP and ATP but an enhanced basal activity and increased phosphorylation of the alpha subunit. Other PRKAG2 missense mutations were previously identified in patients with autosomal dominant hypertrophic cardiomyopathy with Wolff-Parkinson-White syndrome, characterized by juvenile-to-adult clinical onset, moderate cardiac glycogenosis, disturbed excitation conduction, risk of sudden cardiac death in midlife, and molecular perturbations that are similar to--but less severe than- those observed for the R531Q mutation. Thus, recurrent heterozygous R531Q missense mutations in PRKAG2 give rise to a massive nonlysosomal cardiac glycogenosis of fetal symptomatic onset and rapidly fatal course, constituting a genotypically and clinically distinct variant of hypertrophic cardiomyopathy with Wolff-Parkinson-White syndrome. R531Q and other PRKAG2 mutations enhance the basal activity and alpha -subunit phosphorylation of AMP-activated protein kinase, explaining the dominant nature of PRKAG2 disease mutations. Since not all cases displayed PRKAG2 mutations, fatal congenital nonlysosomal cardiac glycogenosis seems to be genetically heterogeneous. However, the existence of a heart-specific primary phosphorylase kinase deficiency is questionable, because no phosphorylase kinase mutations were found. PMID- 15877280 TI - Replication of autism linkage: fine-mapping peak at 17q21. AB - Autism is a heritable but genetically complex disorder characterized by deficits in language and in reciprocal social interactions, combined with repetitive and stereotypic behaviors. As with many genetically complex disorders, numerous genome scans reveal inconsistent results. A genome scan of 345 families from the Autism Genetic Resource Exchange (AGRE) (AGRE_1), gave the strongest evidence of linkage at 17q11-17q21 in families with no affected females. Here, we report a full-genome scan of an independent sample of 91 AGRE families with 109 affected sibling pairs (AGRE_2) that also shows the strongest evidence of linkage to 17q11 17q21 in families with no affected females. Taken together, these samples provide a replication of linkage to this chromosome region that is, to our knowledge, the first such replication in autism. Fine mapping at 2-centimorgan (cM) intervals in the combined sample of families with no affected females reveals a linkage peak at 66.85 cM, which places this locus at 17q21. PMID- 15877281 TI - Identification of FOXP2 truncation as a novel cause of developmental speech and language deficits. AB - FOXP2, the first gene to have been implicated in a developmental communication disorder, offers a unique entry point into neuromolecular mechanisms influencing human speech and language acquisition. In multiple members of the well-studied KE family, a heterozygous missense mutation in FOXP2 causes problems in sequencing muscle movements required for articulating speech (developmental verbal dyspraxia), accompanied by wider deficits in linguistic and grammatical processing. Chromosomal rearrangements involving this locus have also been identified. Analyses of FOXP2 coding sequence in typical forms of specific language impairment (SLI), autism, and dyslexia have not uncovered any etiological variants. However, no previous study has performed mutation screening of children with a primary diagnosis of verbal dyspraxia, the most overt feature of the disorder in affected members of the KE family. Here, we report investigations of the entire coding region of FOXP2, including alternatively spliced exons, in 49 probands affected with verbal dyspraxia. We detected variants that alter FOXP2 protein sequence in three probands. One such variant is a heterozygous nonsense mutation that yields a dramatically truncated protein product and cosegregates with speech and language difficulties in the proband, his affected sibling, and their mother. Our discovery of the first nonsense mutation in FOXP2 now opens the door for detailed investigations of neurodevelopment in people carrying different etiological variants of the gene. This endeavor will be crucial for gaining insight into the role of FOXP2 in human cognition. PMID- 15877282 TI - Deficiency of the ADP-forming succinyl-CoA synthase activity is associated with encephalomyopathy and mitochondrial DNA depletion. AB - The mitochondrial DNA (mtDNA) depletion syndrome is a quantitative defect of mtDNA resulting from dysfunction of one of several nuclear-encoded factors responsible for maintenance of mitochondrial deoxyribonucleoside triphosphate (dNTP) pools or replication of mtDNA. Markedly decreased succinyl-CoA synthetase activity due to a deleterious mutation in SUCLA2, the gene encoding the beta subunit of the ADP-forming succinyl-CoA synthetase ligase, was found in muscle mitochondria of patients with encephalomyopathy and mtDNA depletion. Succinyl-CoA synthetase is invariably in a complex with mitochondrial nucleotide diphosphate kinase; hence, we propose that a defect in the last step of mitochondrial dNTP salvage is a novel cause of the mtDNA depletion syndrome. PMID- 15877283 TI - Alterations in thigh subcutaneous adipose tissue gene expression in protease inhibitor-based highly active antiretroviral therapy. AB - Use of protease inhibitor (PI)-based highly active antiretroviral therapy (HAART) has been associated with altered regional fat distribution, insulin resistance, and dyslipidemias. To assess how PI-based HAART affects adipocyte gene expression in male HIV-1-infected patients, reverse transcription-polymerase chain reaction was used to quantify messenger RNA expression of adipocyte transcription factors and adipocytokines in thigh and abdominal subcutaneous adipose tissue from male (1) HIV-1 seronegative subjects (control, n = 9), (2) asymptomatic treatment naive HIV-1-infected patients (naive, n = 6), (3) HIV-1-infected patients who were receiving antiretroviral agents but never received PIs (PI naive, n = 5), (4) HIV-1-infected patients who were receiving PI-based HAART (PI, n = 7), and (5) HIV-1-infected patients who discontinued the PI component of their antiviral therapy more than 6 months before enrollment (past PI, n =7). In the PI group, the messenger RNA expression levels of the CCAAT/enhancer-binding protein alpha , leptin, and adiponectin (18%, P < .01; 23%, P < .05; and 13%, P < .05, respectively) were significantly lower than the levels measured in the PI-naive group. These results are consistent with previous studies on the effects of PIs on cultured adipocytes. Prospective longitudinal studies of thigh fat adipose tissue gene expression could provide further insights on the pathogenesis of metabolic complications associated with PI-based HAART. PMID- 15877284 TI - Association between DNA variant sites in the apolipoprotein A5 gene and coronary heart disease in Chinese. AB - The recently discovered apolipoprotein A5 ( APOA5 ) gene has been shown to be important in determining plasma triglyceride levels, a major cardiovascular disease risk factor. We searched for possible associations of the APOA5 gene polymorphisms S19W and -1131T>C with coronary heart disease (CHD) in a Chinese population. A total of 483 Chinese CHD patients and 502 control non-CHD subjects were genotyped by polymerase chain reaction-restriction fragment length polymorphism for these 2 single nucleotide polymorphisms. We found that the minor allele 19W was observed only in CHD patients and not in controls, with allelic frequencies of 0.047 and 0.000, respectively ( P < .000001), and the minor allele -1131C was significantly higher in CHD patients than in controls (0.391 vs 0.299, P < .0001). These results suggest that both the S19W and -1131T>C variations in the APOA5 gene are associated with the CHD and appear to be 2 genetic risk factors for CHD susceptibility in Chinese. Moreover, we found that triglyceride levels were significantly higher in -1131C carriers than in -1131T subjects of the control group and that high-density-lipoprotein cholesterol was decreased in 1131C carriers among CHD patients. PMID- 15877285 TI - A retinoid X receptor antagonist, HX531, improves leptin resistance without increasing plasma leptin level in KK-Ay mice under normal dietary conditions. AB - 4-(5 H -2,3-(2,5-Dimethyl-2,5-hexano)-5-methyl-8-nitrodibenzo[b,e][1,4]diazepin 11-yl)benzoic acid (HX531) is a novel retinoid X receptor antagonist. This study provides evidence that HX531 improves leptin resistance without increasing plasma leptin levels in KK-A y mice, an animal model with high plasma leptin levels and leptin resistance. Under normal dietary conditions, 3 weeks of treatment with HX531 (0.03% and 0.06% food admixture) in KK-A y mice decreased plasma leptin levels in a dose- and time-dependent manner, in addition to decreasing body weight and mesenteric fatty tissue weight. To evaluate the effect of HX531 on leptin resistance, leptin was injected intraperitoneally in the KK-A y mice for 4 days after 1 week of treatment with HX531 (0.06% food admixture). This pretreatment with HX531 resulted in exogenously administered leptin causing a significant decrease in food intake. These results suggested that HX531 decreased plasma leptin levels accompanied by a decrease in fatty tissue content in the KK A y mice and a simultaneous improvement in leptin resistance. This is the first report that HX531 improves leptin resistance without increasing plasma leptin level in KK-A y mice, under normal dietary conditions. PMID- 15877286 TI - The prandial insulin sensitivity-modifying effect of vagal stimulation in rats. AB - The effect of left cervical vagal nerve stimulation was studied on insulin sensitivity to test the proposed permissive insulin-sensitizing role of hepatic vagal parasympathetic efferent pathways in fasted and fed anesthetized rats. In fed animals, electrical stimulation (square impulses: 25 V, 5 Hz, 0.5 milliseconds over 15 minutes) of the vagal nerve induced hyperglycemia and an increase in plasma insulin immunoreactivity. Atropine (1.0 mg/kg intravenously) induced insulin resistance estimated by rapid insulin sensitivity testing. This was amplified when the vagal nerve was stimulated. The insulin-resistant state developed by fasting was not modified by either treatment with atropine or electrical stimulation. We conclude that both parasympathetic cholinergic and noncholinergic vagal efferents modulate postprandial neurogenic insulin sensitivity adjustments. PMID- 15877287 TI - Corticosteroid-binding globulin affects the relationship between circulating adiponectin and cortisol in men and women. AB - Inflammatory pathways are increasingly recognized to be tightly associated with insulin resistance in humans. The promoter region of the adiponectin gene--Apm1- encompasses consensus sequences for glucocorticosteroid receptor responsive element. Dexamethasone induced downregulation of adiponectin secretion in vitro, whereas prednisolone administration increased circulating adiponectin concentrations. As previous studies have demonstrated an inverse relationship between corticosteroid-binding globulin (CBG), body mass index, and insulin resistance, we studied whether CBG could explain cortisol-to-adiponectin relationship. One hundred twenty-two healthy subjects were enrolled in a cross sectional study. Plasma CBG and serum cortisol concentration were measured by radioimmunoassay. The cortisol-to-CBG ratio was used to calculate free cortisol. An RIA kit (Linco Research, St Louis, MO) was used to measure adiponectin levels. Insulin resistance was calculated using the homeostatis model of assessment (HOMA) value. Circulating adiponectin was associated with serum CBG ( r = 0.38, P < .00001), both in men ( r = 0.26, P = .03, n = 79) and women ( r = 0.48, P = .003, n = 43), and with insulin resistance (HOMA index) ( r = -0.30, P < .0001) in both. Free cortisol correlated negatively with adiponectin only in women ( r = -0.32, P = .04), but not in men ( r = 0.01, P = .89). Serum CBG concentration was significantly lower among men in the lowest quartile of adiponectin when compared with the remaining subjects (37.3 +/- 5.7 vs 40.6 +/- 5.1, P = .016), whereas men in the highest quartile of adiponectin showed significantly increased free cortisol index (14.2 +/- 3.3 vs 12.2 +/- 3.1, P = .039). Women in the lowest quartile of adiponectin also displayed significantly lower CBG concentration than that present in the remaining subjects (38.6 +/- 6.9 vs 44.4 +/- 5.5, P = .016), whereas free cortisol index was not significantly different across adiponectin quartiles ( P = .1). In a stepwise regression analysis, body mass index ( P = .0011), CBG ( P = .0047), and sex ( P = .04) contributed to 15%, 8%, and 3%, respectively, of adiponectin variance. Using CBG as dependent variable, both adiponectin ( P = .0002) and fasting cortisol ( P = .019) contributed to 14% and 4%, respectively, of CBG variance. In summary, circulating adiponectin, CBG concentration, and fasting cortisol were significantly interrelated in healthy subjects. A significant sexual dimorphism exists in this association. PMID- 15877288 TI - Peroxisome proliferator-activated receptor-gamma agonist rosiglitazone reduces clinical inflammatory responses in type 2 diabetes with coronary artery disease after coronary angioplasty. AB - Rosiglitazone, an agonist of peroxisome proliferator-activated receptor-gamma (PPAR gamma ), is an insulin-sensitizing antidiabetic agent and inhibits restenosis in animal blood vessels. However, its benefit for patients with type 2 diabetes and coronary artery disease (CAD) after percutaneous coronary intervention is unknown. Patients with diabetes and CAD who had undergone percutaneous coronary intervention were randomized to either receive or not receive rosiglitazone (4 mg/d) for 6 months. After 6 months of rosiglitazone treatment, the plasma levels of fasting glucose and insulin and those of hemoglobin A1C and homeostasis model assessment of insulin resistance were significantly decreased in the rosiglitazone group as compared with baseline levels and those in the control group. After 2 and 6 months of rosiglitazone treatment, the plasma level of high-density lipoprotein was significantly increased in the rosiglitazone group. In addition, plasma levels of monocyte chemoattractant protein-1 and C-reactive protein and hyperresponsiveness of low dose lipopolysaccharide-induced monocyte chemoattractant protein-1 secretion from monocytes were reduced. Furthermore, the occurrence of coronary events was significantly decreased in the rosiglitazone group at 6-month follow-up. Our data indicate that rosiglitazone may protect the vascular wall through not only improving the features of metabolic disorders but also reducing proinflammatory responses and the occurrence of coronary events in patients with diabetes and CAD after percutaneous coronary intervention. PMID- 15877289 TI - Analysis of insulin-stimulated insulin receptor activation and glucose transport in cultured skeletal muscle cells from obese subjects. AB - Obesity is associated with impaired insulin-stimulated glucose disposal in the skeletal muscle, but whether this is an intrinsic or acquired factor is unknown. In many patients with type 2 diabetes mellitus (T2D) and their nondiabetic relatives, who have a genetic predisposition for diabetes, insulin resistance is maintained in cultured muscle cells. To study the association of obesity with defects in insulin action, we investigated insulin stimulation of both insulin receptor (IR) autophosphorylation and subsequent glucose transport in primary skeletal muscle cell cultures obtained from both nonobese and obese nondiabetic subjects. In these 2 groups, there was no difference in the ability of insulin to induce autophosphorylation of the IR, phosphorylation of the downstream serine kinase Akt/PKB, or stimulation of glucose transport. Moreover, there were no major differences in cultured muscle cell content of either the IR, the IR antagonist PC-1, or GLUT 1 and GLUT 4. These data therefore indicate that the insulin resistance associated with obesity is not maintained in cultured muscle cells and suggest that this insulin resistance is an acquired feature of obesity. PMID- 15877290 TI - Growth hormone administration increases glucose production by preventing the expected decrease in glycogenolysis seen with fasting in healthy volunteers. AB - Twelve volunteers were fasted overnight and infused with [ 13 C]glucose (ul) to measure glucose production (GP), gluconeogenesis, and by subtraction, glycogenolysis. Glucose production, gluconeogenesis, and glycogenolysis were measured after a 3-hour baseline infusion and two 4-hour infusions. The first 4 hours of the pituitary-pancreatic clamp study (PPCS) with replacement insulin, cortisol, and glucagon was without growth hormone (GH) administration. The second 4 hours of the PPCS was with high-dose GH administration. Six fasting volunteers acted as controls over the 11-hour study period. Overnight 12-hour fasting measurements of hormones, glucose, GP, gluconeogenesis, and glycogenolysis were similar in both groups. The PPCS had no significant effect on GP (2.43 +/- 0.19 vs 2.07 +/- 0.11 mg/kg per minute, PPCS vs controls, mean +/- SEM). Glycogenolysis, as a percent of GP (43%-49%), was similar between PPCS and controls (43% +/- 3% vs 49% +/- 4%). High-dose GH for 4 hours increased GH (20.8 +/- 3.8 vs 2.0 +/- 0.9 ng/mL), blood glucose (127 +/- 28 vs 86 +/- 4 mg/dL, P < .05), GP (2.21 +/- 0.21 vs 1.81 +/- 0.12 mg/kg per minute, P < .05). The increase in GP was due to sustained glycogenolysis as compared to the observed fall in glycogenolysis seen with fasting alone (0.94 +/- 0.21 vs 0.53 +/- 0.07 mg/kg per minute, P < .05). Glycogenolysis, as a percent of GP, was significantly increased with high-dose GH (43 +/- 5% vs 29 +/- 3%, P < .05). High-dose GH had no effect on gluconeogenesis (1.26 +/- 0.15 vs 1.29 +/- 0.12 mg/kg per minute). High-dose GH prevents the fall in glycogenolysis observed with fasting alone. PMID- 15877291 TI - Oxidation of combined ingestion of glucose and sucrose during exercise. AB - The first purpose of the study was to examine whether combined ingestion of glucose and sucrose at an intake rate of 1.2 g/min would lead to higher oxidation rates compared with the ingestion of an isocaloric amount of glucose or sucrose alone. The second aim of the study was to investigate whether a mixture of glucose and sucrose when ingested at a high rate (2.4 g/min) would result in exogenous CHO oxidation rates higher than 1.2 to 1.3 g/min. Eight trained cyclists (maximal oxygen consumption: 64 +/- 2 mL . kg -1 . min -1 , mean +/- SE) performed 5 exercise trials in random order. Each trial consisted of 120 minutes of cycling at 50% maximum power output (63% +/- 2% maximal oxygen consumption), whereas subjects received a solution providing either 1.2 g/min of glucose (GLU), 1.2 g/min of sucrose (SUC), 0.6 g/min of glucose + 0.6 g/min of sucrose (M GLU+SUC), 1.2 g/min of glucose + 1.2 g/min of sucrose (H-GLU+SUC), or water (WAT). Peak exogenous CHO oxidation rates in the H-GLU+SUC trial (1.20 +/- 0.07 g/min) were significantly higher ( P < .01) compared with the GLU, M-GLU+SUC, and SUC trials (0.77 +/- 0.04, 0.90 +/- 0.07, 0.98 +/- 0.04 g/min, respectively). Furthermore, peak exogenous CHO rates in M-GLU+SUC and SUC trials were significantly higher ( P < .05) compared with the GLU trial. In conclusion, combined ingestion of moderate amounts of glucose and sucrose (144 g) during cycling exercise resulted in approximately 21% higher exogenous CHO oxidation rates compared with the ingestion of an isocaloric amount of glucose. Furthermore, when a mixture of glucose and sucrose was ingested at high rates (2.4 g/min), exogenous CHO oxidation rates reached peak values of approximately 1.20 g/min. PMID- 15877292 TI - Synthetic protease inhibitor camostat prevents and reverses dyslipidemia, insulin secretory defects, and histological abnormalities of the pancreas in genetically obese and diabetic rats. AB - BACKGROUND: Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a model of type 2 diabetes, lacks the expression of cholecystokinin-1 receptor mRNA and exhibits inflammation and degeneration of the pancreas and eventually develops insulinopenic diabetes. Protease inhibitors are known to modulate inflammatory response and fibrosis as well as inhibit proteases activity. AIM: To examine the effects of long-term treatment with camostat, a synthetic protease inhibitor, on metabolic and histopathological changes in the islets of OLETF rats. METHOD: OLETF rats were fed either camostat-containing food (200 mg/100 g) from 12 or 28 weeks of age to 72 weeks of age, or fed standard rat diet. RESULTS: Camostat-fed rats gained less weight or lost weight, although they consumed more food than the control rat when food intake was adjusted for body weight. Camostat reduced visceral adipose depots and fasting serum concentrations of triglyceride, free fatty acids, cholesterol, glucose, and insulin. Pancreatic insulin content in camostat-treated rats was significantly higher than in control rats. Immunohistochemistry revealed marked suppression of expressions of tumor necrosis factor alpha , interleukin 1 beta , interleukin 6, and alpha-smooth muscle actin in the islets of camostat-treated rats, compared with control rats. Histologically, disruption of the islets and pancreatic fibrosis were noted in control rats but not in camostat-fed rats. CONCLUSION: Our findings suggest that camostat prevents and reverses obesity, hyperinsulinemia, hyperglycemia, and hyperlipidemia and markedly inhibits inflammation, fibrosis, and disruption of the islets in the genetically obese diabetic OLETF rats. PMID- 15877293 TI - Functional polymorphism in Z-DNA-forming motif of promoter of SLC11A1 gene and type 1 diabetes in Japanese subjects: association study and meta-analysis. AB - The association of the polymorphism of the Z-DNA-forming repeats in the promoter region of SLC11A1 (solute carrier family 11 member 1), formerly designated NRAMP1 (natural resistance associated macrophage protein 1), with type 1 diabetes was studied in a total of 244 Japanese subjects. Three alleles were detected in Japanese subjects. In diabetic patients, allele 2 was less frequent and allele 3 was more frequent, albeit not significantly, than in control subjects. Allele 2 was significantly ( P < .024) less frequent whereas allele 3 was more, albeit not significantly, frequent in the younger onset group than in the control subjects. In patients with a susceptible HLA allele, DRB1*0405 or DRB1*0901 , the frequency of allele 2 was significantly ( P < .013) lower and that of allele 3 tended to be higher than that in patients without either DRB1*0405 or DRB1*0901 . The protective effect of allele 2 against type 1 diabetes and other autoimmune diseases was confirmed by meta-analysis (summary odds ratio, 0.71, 95% confidence interval, 0.53-0.96). Because allele 2 was shown to be associated with low expression of SLC11A1 and protection against another autoimmune disease, rheumatoid arthritis, the negative association of allele 2 with autoimmune type 1 diabetes in the present study suggests that a less active immune system in subjects with allele 2 may protect individuals from autoimmune diseases. PMID- 15877294 TI - Role of hypoxia-induced anorexia and right ventricular hypertrophy on lactate transport and MCT expression in rat muscle. AB - To dissect the independent effects of altitude-induced hypoxemia and anorexia on the capacity for cardiac lactate metabolism, we examined the effects of 21 days of chronic hypobaric hypoxia (CHH) and its associated decrease in food intake and right ventricle (RV) hypertrophy on the monocarboxylate transporter 1 and 4 (MCT) expression, the rate of lactate uptake into sarcolemmal vesicles, and the activity of lactate dehydrogenase isoforms in rat muscles. In comparison with control rats (C), 1 mmol/L lactate transport measured on skeletal muscle sarcolemmal vesicles increased by 33% and 58% in hypoxic (CHH, barometric pressure = 495 hPa) and rats pair-fed an equivalent quantity of food to that consumed by hypoxic animals, respectively. The increased lactate transport was higher in PF than in CHH animals ( P < .05). No associated change in the expression of MCT1 protein was observed in skeletal muscles, whereas MCT1 mRNA decreased in CHH rats, in comparison with C animals (42%, P < .05), partly related to caloric restriction (30%, P < .05). MCT4 mRNA and protein increased during acclimatization to hypoxia only in slow-oxidative muscles (68%, 72%, P < .05, respectively). The MCT4 protein content did not change in the plantaris muscle despite a decrease in transcript levels, related to hypoxia and caloric restriction. In both the left and right ventricles, the MCT1 protein content was unaffected by ambient hypoxia or restricted food consumption. These results suggest that MCT1 and MCT4 gene expression in fast-glycolytic muscles is mainly regulated by posttranscriptional mechanisms. Moreover, the results emphasize the role played by caloric restriction on the control of gene expression in response to chronic hypoxia and suggest that hypoxia-induced right ventricle hypertrophy failed to alter MCT proteins. PMID- 15877295 TI - Rosiglitazone reduces serum homocysteine levels, smooth muscle proliferation, and intimal hyperplasia in Sprague-Dawley rats fed a high methionine diet. AB - Homocysteine (Hcy) is a metabolite of the essential amino acid methionine. Hyperhomocysteinemia is associated with vascular disease, particularly carotid stenosis. Rosiglitazone, a ligand of the peroxisome proliferator-activated receptor gamma , attenuates balloon catheter-induced carotid intimal hyperplasia in type 2 diabetic rats. We studied 4 groups (n = 7 per group) of adult female Sprague-Dawley rats fed (a) powdered laboratory chow (control), (b) control diet with rosiglitazone (3.0 mg/kg/d), (c) diet containing 1.0% l -methionine, and (d) diet containing methionine and rosiglitazone. After 1 week on high methionine diet, the rats were administered an aqueous preparation of rosiglitazone by oral gavage. One week after initiation of rosiglitazone, balloon catheter injury of the carotid artery was carried out using established methods, and the animals continued on their respective dietary and drug regimens for another 21 days. At the end of the experimental period, blood samples were collected, and carotid arteries and liver were harvested. Serum Hcy increased significantly on methionine diet compared with controls (28.9 +/- 3.2 vs 6.3 +/- 0.04 micromol/L). Development of intimal hyperplasia was 4-fold higher in methionine-fed rats; this augmentation was significantly reduced ( P < .018) in rosiglitazone-treated animals. Rosiglitazone treatment significantly ( P < .001) suppressed Hcy levels and increased the activity of the Hcy metabolizing enzyme, cystathionine-beta synthase in the liver samples. Hcy (100 micromol/L) produced a 3-fold increase in proliferation of rat aortic vascular smooth muscle cells; this augmentation was inhibited by incorporating rosiglitazone (10 micromol/L). After balloon catheter injury to the carotid artery of animals on a high methionine diet, there was an increase in the rate of development of intimal hyperplasia consistent with the known effects of Hcy. It is demonstrated for the first time that the peroxisome proliferator-activated receptor gamma agonist rosiglitazone can attenuate the Hcy stimulated increase in the rate of development of intimal hyperplasia indirectly by increasing the rate of catabolism of Hcy by cystathionine-beta-synthase and directly by inhibiting vascular smooth muscle cell proliferation. These findings may have important implications for the prevention of cardiovascular disease and events in patients with hyperhomocysteinemia (HHcy). PMID- 15877296 TI - Close association of hypoadiponectinemia with arteriosclerosis obliterans and ischemic heart disease. AB - Adiponectin is an adipose-derived cytokine, and it is suggested that hypoadiponectinemia increases the prevalence of ischemic heart disease (IHD). The present study was undertaken to determine serum adiponectin levels in patients with arteriosclerosis obliterans (ASO) and IHD. Forty-nine patients with ASO and 49 age-, sex-, and body mass index-matched control subjects were examined. The diagnosis of ASO was derived from an ankle brachial index of less than 0.90 and stenotic or obstructive change in angiogram. Ischemic heart disease was diagnosed by ischemic or stenotic change in ECG, treadmill, or coronary angiogram. Serum adiponectin level was 8.6 +/- 0.9 microg/mL in the patients with ASO, a value significantly less than that of 12.4 +/- 1.0 microg/mL in the control subjects ( P < .01). Next, we subgrouped the subjects into 4 groups according to the presence of ASO and IHD. Serum adiponectin levels were 9.4 +/- 1.5 and 10.2 +/- 1.6 microg/mL in the subjects with ASO (n =23) and those with IHD (n = 13), respectively. It was further reduced to 7.9 +/- 1.2 microg/mL in the subjects having both ASO and IHD (n = 26), a value significantly less than that of 13.2 +/ 1.4 microg/mL in the control subjects (n = 36; P < .05). Serum high-density lipoprotein cholesterol was significantly less in the subjects with ASO than in the control subjects (42.1 +/- 1.7 vs 48.5 +/- 2.0 mg/dL; P < .05), but there were no differences in blood pressure, total cholesterol, low-density lipoprotein cholesterol, triglyceride, and uric acid levels. The present results indicate that a reduction in serum adiponectin level is associated with the prevalence and magnitude of systemic atherosclerosis including IHD and ASO. PMID- 15877297 TI - Cortitrol supplementation reduces serum cortisol responses to physical stress. AB - The supplement Cortitrol was formulated to mitigate the cortisol response to physiological and psychological stress. Therefore, the purpose of this study was to examine the effects of Cortitrol on serum cortisol concentrations before, during, and after a high-intensity resistance exercise protocol (EX) and a resting control day (REST). We used a matched, balanced, randomized, double blind, placebo-controlled, cross-over design. Blood samples were obtained at matching time points during EX and REST. Cortitrol significantly ( P < .05) reduced cortisol area under the curve concentrations during REST. During EX, Cortitrol reduced cortisol concentrations at 20, 10, and 0 minutes pre-exercise, at mid-exercise, immediately post-exercise, and at 5 minutes post-exercise. In addition, serum cortisol and plasma adrenocorticotropin hormone area under the curve concentrations during EX were significantly lower after Cortitrol than placebo. Furthermore, Cortitrol significantly reduced free radical production. This was indicated by significantly lower plasma malondialdehyde concentrations at the 65-minute post-exercise time point during REST, and at pre-exercise, immediate post-exercise, and 65 minutes post-exercise during EX. Serum total testosterone, free testosterone, dehydroepiandrosterone, and growth hormone showed exercise-induced increases but no treatment effects. These data demonstrate that Cortitrol was effective in modulating the physiological stress responses of exercise from the anticipatory rises before physical stress and into early recovery by reducing cortisol and associated free radical production. PMID- 15877298 TI - Association of decrease in serum dehydroepiandrosterone sulfate levels with the progression to type 2 diabetes in men of a Japanese population: the Funagata Study. AB - Association of serum dehydroepiandrosterone sulfate (DHEAS) levels with insulin resistance and impairment of insulin secretion have been reported. We here examined the association of serum DHEAS levels with type 2 diabetes mellitus (DM) and the progression to DM. The serum DHEAS levels at baseline (from 1995 to 1997) were evaluated in 1709 individuals (998 women and 711 men) from a cohort population (n = 3706) of the Funagata Study. Glucose tolerance was evaluated at baseline as well as at 5-year follow-up examinations (n = 970, follow-up rate, 56.8%) according to the 1985 World Health Organization criteria. The statistical significance of the difference between any 2 groups was determined by the Student t test. Multiple logistic regression analysis determined the association of the traits with the progression to DM at the 5-year follow-up examinations. P < .05 was accepted as statistically significant. The serum DHEAS levels were significantly lower in DM than in normal glucose tolerance. However, this difference was not significant when adjusted for age. In men, the decrease in serum DHEAS levels by the 5-year follow-up examinations was significantly larger in the subjects who became diabetic than in the subjects who remained normal glucose tolerance, even when adjusted for age ( P = .0003). Multiple logistic regression analysis revealed a significant association of the decrease in serum DHEAS levels with the progression to DM, with an odds ratio (per 0.1 log ng/mL) of 1.410 (95% confidence interval [CI], 1.020-1.948, P = .038), independently from age, height, and 2-hour plasma glucose in men. A decrease in serum DHEAS levels seems to be associated with the progression to DM in Japanese men. PMID- 15877299 TI - The beneficial effects of lipid-lowering drugs beyond lipid-lowering effects: a comparative study with pravastatin, atorvastatin, and fenofibrate in patients with type IIa and type IIb hyperlipidemia. AB - Hyperlipidemia is an important risk factor for atherosclerosis. Hemorheological factors contribute to morbidity and mortality in patients with dyslipidemia. We evaluated the effects of 3 antihyperlipidemic drugs (pravastatin, atorvastatin, and fenofibrate), which have different mechanisms of action and different patterns of action on lipid profiles, on erythrocyte deformability and fibrinogen levels in patients with type IIa and type IIb hyperlipidemia. Twenty-one patients ( 4 men and 17 women) with type IIa and IIb hyperlipidemia were randomized to 3 drugs (pravastatin 20 mg/d, atorvastatin 10 mg/d, fenofibrate 250 mg/d) for 8 weeks. Plasma glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) analysis were performed on a BM-Hitachi 747-200 autoanalyzer (Hitachi-Roche, Tokyo, Japan). Fibrinogen analysis was performed according to Clauss method. Erythrocyte deformability was assessed with cell transit analysis device. There was no significant difference in body mass index, lipid profile, fibrinogen level, and erythrocyte deformability index values among the groups before treatment ( P > .05). In all groups, there were statistically significant reductions in total LDL C levels ( P < .05). The triglyceride levels were significantly reduced in the atorvastatin and fenofibrate groups ( P < .05), but not in the pravastatin group ( P > .05). There was no significant change in HDL-C levels during the treatment with statins ( P > .05), but there was a significant increase in the fenofibrate group ( P < .05). Mean erythrocyte deformability index was improved in all the groups ( P < .05). There was no significant change in fibrinogen levels during the treatment of pravastatin and atorvastatin ( P > .05), but in fenofibrate group, fibrinogen levels were significantly decreased ( P < .05). The 3 groups of antihyperlipidemic drugs have beneficial effects on the erythrocyte deformability index. Only fenofibrate has significant beneficial effects on the fibrinogen levels. PMID- 15877300 TI - Absence of fatty liver in familial hypobetalipoproteinemia linked to chromosome 3p21. AB - Our aim was to ascertain whether fatty liver may be present in the genetic form of familial hypobetalipoproteinemia (FHBL) linked to a susceptibility locus on chromosome 3p21. Three genetic forms of FHBL exist: (a) FHBL caused by truncation specifying mutations of apolipoprotein B (apoB), (b) FHBL linked to chr3p21, and (c) FHBL not linked either to APOB or to chr3p21. Fatty liver is common in apoB defective FHBL. Hepatic fat contents were quantified by magnetic resonance spectroscopy in 16 subjects with 3p21-linked FHBL, 32 subjects with apoB defective FHBL, and 39 sex- and age-matched controls. Mean liver fat of 3p21 subjects was similar to controls and approximately 60% lower than apoB-defective FHBL subjects ( P = .0012). Indices of adiposity (body mass index, waist/hip ratio) and masses of abdominal subcutaneous, retroperitoneal, and intraperitoneal adipose tissue (IPAT) were quantified by MR imaging. Mean measures of adiposity were similar in the 3 groups, suggesting that adiposity per se was not responsible for differences in liver fat. Liver fat content was positively correlated with IPAT. The intercepts of regression lines of IPAT on liver fat content were similar in controls and 3p21, but higher in apoB-defective FHBL subjects. The slopes of the lines were steepest in apoB-defective, intermediate in 3p21, and flattest in controls. Lipoprotein profiles and very low density lipoprotein-apoB100 kinetics of 3p21 and apoB-defective groups also differed. Thus, 2 genetic subtypes of FHBL also differ in several phenotypic features. PMID- 15877301 TI - Intestinal assembly and secretion of highly dense/lipid-poor apolipoprotein B48 containing lipoprotein particles in the fasting state: evidence for induction by insulin resistance and exogenous fatty acids. AB - Emerging evidence suggests that overproduction of intestinally derived apolipoprotein (apo) B48-containing lipoprotein particles may be an important contributor to both fasting and postprandial dyslipidemia in insulin-resistant states. Mechanisms regulating the assembly and secretion of apoB48-containing lipoproteins are not fully understood particularly in the diabetic/insulin resistant intestine. In the present study, we have investigated the density profile of apoB48 lipoproteins assembled in primary hamster enterocytes. Both intracellular and secreted apoB48 particles were examined in intestinal enterocytes isolated from normal or insulin-resistant fructose-fed hamsters, as well as in enterocytes treated with exogenous oleic acid. Microsomal luminal contents and culture media were analyzed by discontinuous and sequential ultracentrifugation on sucrose and KBr gradients, respectively. ApoB48 was mostly secreted on VLDL-, LDL-, and denser HDL-sized particles in the fasting state. In pulse-chase labeling experiments, nascent apoB48-containing particles initially accumulated in the microsomal lumen as HDL-sized particles, with subsequent formation of apoB48-VLDL particles, with only a minute amount of chylomicrons observed. Treatment with 720 mu mol/L of oleic acid, increased microsomal apoB48 HDL synthesis, and induced a marked shift toward lighter more buoyant particles. A marked enhancement in assembly of apoB48-containing lipoproteins was also observed in the microsomal lumen of fructose-fed hamster enterocytes, suggesting facilitated assembly and secretion of dense intestinal lipoprotein particles in insulin-resistant states. Overall, these observations suggest that a major proportion of apoB48-containing lipoprotein particles is assembled and secreted as highly dense, HDL-sized particles. The production of these small, dense, and potentially atherogenic apoB48 particles can be stimulated by increased free fatty acid flux as well as in insulin-resistant diabetes. PMID- 15877305 TI - Overview of the field. AB - Clinical descriptions of depression date back to antiquity. Scientific investigations of mood disorders started only 150 years ago. An historical overview of depression leads into today's knowledge that depressive disorders are common, serious, and sometimes life-threatening and that their effects are persistent and costly. Depressive disorders have a high prevalence, around 5% in the general population, with at least 20% of patients suffering with chronic conditions, such as cardiovascular disease and diabetes. Neuroimaging technology provides unprecedented opportunities for elucidating the neurobiological correlates of mood disorders. Neuroimaging studies of primary mood disorders have identified neurophysiologic abnormalities in the orbital and medial prefrontal cortex (PFC), the amygdala, and the related parts of the striatum and thalamus. There has been a revolution in our understanding of the pathogenesis of depression, with recent work demonstrating, among others, the critical impact of stress. There is an abundant evidence from family, twin, and adoption studies that genetic factors play an important role in the etiology of affective disorders. A variety of hormonal abnormalities, such as altered levels of cortisol, growth hormone, or thyroid hormones, indicate the existence of endoctrine disturbances. Despite the initial findings of immunosuppresion in depression, some studies have indicated that immune activation could also be present and might even play a role in the onset of depressive symptoms. Neurotrophic factors are among the growth factors that have been studied for their role in the adult nervous system. Despite advances in the pharmacotherapy of depression, only one third of patients respond favorably to antidepressant drugs. One third do not respond at all, and in clinical trials, at least one third respond to placebo. There is clearly an urgent need for novel antidepressants. PMID- 15877306 TI - Genetics and genomics of depression. AB - Depressive disorders are among the most common psychiatric diseases, with prevalence estimates ranging from 5% to a maximum of 20%. Despite their high prevalence and socioeconomic impact, little is known about their etiology. Heritability estimates demonstrate up to a 50% genetic component based on family aggregation and contrasting monozygotic and dizygotic twin studies. The low relative risk to siblings ( lambda sib <1.5) makes the search for their genetic determinants very tedious. Gene-environment interaction has been recognized for a long time in the pathophysiology of depression, and its best biological substratum at present is represented by the serotonin transporter (5-HTT) gene, where several copies of its short allele culminate in depression and suicide in response to lifelong stress events. Many total genome scans have been performed with variable results, the most authoritative being the one of Utah pedigrees with a strong family history of major depression. It identified a locus on chromosome 12 encompassing a gene cluster and sex-specific predisposition. Nevertheless, recent genome scan meta-analysis yielded somewhat disappointing conclusions with a relatively low significance for quantitative trait loci on chromosomes 9, 10, 14, and 18. Studies on animal models have contributed to the chromosomal mapping of many behavioral traits, including anxiety, the stress response, and depression. Although F2 crosses constitute a classical approach, novel models of recombinant inbred strain and recombinant congenic strain animals allow for a rapid initial localization of any traits. This type of analysis has led us to uncover significant loci for the stress response and anxiety in rats and mice. PMID- 15877307 TI - Genes, stress, and depression. AB - A relationship between genetic makeup and susceptibility to major depressive disorder (MDD) has long been suspected on the basis of family and twin studies. A metaanalysis of reports on the basis of twin studies has estimated MDD's degree of heritability to be 0.33 (confidence interval, 0.26-0.39). Among families exhibiting an increased prevalence of MDD, risk of developing the illness was enhanced in members exposed to a highly stressful environment. Aberrant genes can predispose to depression in a number of ways, for example, by diminishing production of growth factors that act during brain development. An aberrant gene could also increase or decrease a neurotransmitter's release into synapses, its actions, or its duration of activity. The gene products of greatest interest at present are those involved in the synthesis and actions of serotonin; among them, the serotonin-uptake protein localized within the terminals and dendrites of serotonin-releasing neurons. It has been found that the Vmax of platelet serotonin uptake is low in some patients with MDD; also, Vmax is highly correlated in twins. Antidepressant drugs such as the selective serotonin reuptake inhibitors act on this uptake protein. The specific genetic locus causing serotonin uptake to be lower in some patients with major depression involves a polymorphic region (5-HTTLPR) in the promoter region of the gene for the uptake protein. The gene itself exists as several alleles, the short "S" allele and the long "L" allele. The S variant is associated with less, and the L variant with more, of the uptake protein. The effect of stressful life events on depressive symptoms in young adults was found to be significantly stronger among SS or SL subjects than among LL subjects. Neuroimaging studies showed that people with the SS or SL alleles exhibited a greater activation of the amygdala in response to fearful stimuli than those with LL. It has been reported recently that mutations in the gene that controls serotonin synthesis in the human brain (tryptophan hydroxylase) also predispose to mood disturbances. It may be asked whether people who lack a psychiatric history should be advised to avoid stressful environments if they are found to carry the SS or SL alleles. PMID- 15877308 TI - Glucocorticoids, depression, and mood disorders: structural remodeling in the brain. AB - The hippocampal formation expresses high levels of adrenal steroid receptors and is a malleable brain structure that is important for certain types of learning and memory. It is also vulnerable to the effects of stress and trauma. The amygdala is an important target of stress and mediates physiological and behavioral responses associated with fear and strong emotions. The prefrontal cortex plays an important role in working memory and executive function and is also involved in extinction of learning. All 3 regions are targets of stress hormones, and stress is known to precipitate and exacerbate mood disorders. In long-term depressive illness, the hippocampus and prefrontal cortex undergo atrophy, whereas the amygdala is hyperactive in anxiety and mood disorders and may undergo a biphasic change in structure--increasing in size in acute depression and shrinking on long-term depression. In animal models of acute and chronic stress, neurons in the hippocampus and prefrontal cortex respond to repeated stress by showing atrophy that leads to memory impairment, whereas neurons in amygdala show a growth response that leads to increased anxiety and aggression. Yet, these are not necessarily "damaged" and may be treatable with the right medications. The mechanisms that distinguish between protection and damage of brain cells from stress are discussed in this context. PMID- 15877309 TI - Neurobiological basis of depression: an update. AB - The past 5 years have seen unprecedented advances in our knowledge about the neurobiology of depression. Significant breakthroughs have been made in genomics, imaging, and the identification of key neural systems involved in cognition, emotion, and behavior. In addition, novel targets have been identified for the development of new pharmacological and behavioral treatments. Genetic variations associated with most mental disorders are being identified, and reliable tests for early detection of risk and disease are now on the horizon. New neurobiological concepts have emerged, as they relate to these advances in mental health research such as the serotonin transporter receptor, a genetic variant of which doubles the risk of depression. Brain neurochemicals, including neurotropic factors (implicated in several mental disorders), and anatomical studies involving imaging of the amygdala and the hippocampus and prefrontal cortex are now at the forefront. Several brain neurotransmitters systems: glutamate, gamma aminobutyric acid, serotonin, norepinephrine, and dopamine have been implicated in depression and mania. These transmitter systems, as well as other neurochemical systems such as membrane-bound signal transduction systems and intracellular signaling systems that modulate gene transcription and protein synthesis, play an important role in the etiology of depression. This new knowledge is expected to provide important clues for the development of selective pharmacological interventions. Neuroimaging studies of depressed patients have shown several abnormalities of regional cerebral blood flow and glucose metabolism--a surrogate of neuronal function--in various brain regions, including the limbic cortex, the prefrontal cortex, the hippocampus, the amygdala, and the anterior cingulate cortex. At this time, a considerable amount of new information is converging--derived from animal models of mood disorders, genetics, basic behavioral research, and neuroscience. It is inevitable that the next step in this progression will be the integration of these basic advances in clinical management and the application of this new information in the context of the depressed patient. PMID- 15877310 TI - Depression and neurosurgery: past, present, and future. AB - Neurosurgery has been used to treat depression since 1935, when open surgery was first used to isolate relatively large areas of the limbic system from the rest of the brain. Soon thereafter, more selective leucotomies were performed based on a growing knowledge of the role played by brain limbic circuitry in processing the emotions. Subsequent discovery of the effectiveness in depression of both electroconvulsive therapy and various pharmacotherapies raised serious doubts about "psychosurgical" treatments, but the introduction of stereotactic techniques revived interest in the selective-lesion, neurobiology-based approach. However, neurosurgery has only come to be regarded as an appropriate treatment of severe depression since Benabid introduced the frequency-dependent chronic electric stimulation technique. Because of its nondestructive nature, this procedure will undoubtedly be favored in the future. One can anticipate that, eventually, frequency-dependent chronic electric stimulation will be complemented by newer techniques such as microdialysis and reverse dialysis, with concomitant functional magnetic resonance imaging and/or positron emission tomography scanning, and the use of chemodes for microinfusion or for in situ insertion of reactivated-stem cells. To optimize success, these modern methods will require a new taxonomy of "depressions" based on up-to-date neurobiological criteria. PMID- 15877311 TI - Depression, cytokines, and glial function. AB - It has been known for some time that cytokines made and released during systemic illness can result in a constellation of symptoms strikingly similar to those observed in depression. The overlap of the symptoms of depression and systemic illness raises the intriguing possibility that cytokines may be involved in the development and maintenance of mood disorders. Cytokines are small ubiquitous pleiotropic molecules that are made and released in response to a variety of stimuli. They have a multitude of actions throughout the body, including actions on the central and peripheral nervous systems. Alterations in the levels of circulating cytokines, especially the key proinflammatory cytokines, interleukin 6 and tumor necrosis factor alpha , have been linked to a variety of disease states including those involving central nervous system depression. In this brief review, epidemiological and clinical data on depression, as well as findings from relevant animal models, are examined for links between cytokine expression and depression. We suggest that glial cells, both as a source and target of cytokines, represent the overlooked targets involved in the etiology of depression. PMID- 15877312 TI - Subsyndromal depression in the elderly: underdiagnosed and undertreated. AB - Major depressive illness is present in about 5.7% of US residents aged>or=65 years, whereas clinically significant nonmajor or "subsyndromal" depression affects approximately 15% of the ambulatory elderly. Risk of developing subsyndromal depression increases as elderly people get older. Because they have numerous distressing ailments, everyday life can be burdensome for many elderly persons. Almost one third of Americans aged 75 years or older rate their health as "fair to poor." Yet, the physical discomforts experienced by so many elderly individuals are unlikely to generate a clinically significant depression unless other ingredients such as loneliness, impairment of mobility, loss of a spouse, a serious financial reverse, and--probably most important--genetic susceptibility are added to the psychophysiological mix. Because depression damages quality of life and is usually eminently treatable, it is essential that physicians and other health professionals be trained to recognize true depression and distinguish it from confounding conditions caused by medications, organic brain disease, or short-term grief reactions. In the medically ill elderly, depressive symptoms may be overlooked because of the assumption that they are a part of the concurrent medical illness. Diagnosis of depression in the elderly can be greatly assisted by use of age-specific screening instruments such as the Geriatric Depression Scale. Ultimately, brain imaging and biochemical and physiological measurements may prove useful in diagnosis. The presence of somatic concomitants of depression such as severe neck and low back pain should alert the clinician to the possibility of an underlying mood disorder. Suicide and suicide attempts occur all too frequently in the depressed elderly; therefore, screening for late life depression is urgently required among the elderly in primary and residential health care settings. PMID- 15877313 TI - Depression and cardiovascular disease: a reciprocal relationship. AB - Until relatively recently, depression has been considered a purely "mental" disorder and therefore in the natural domain of psychologists and psychiatrists. However, recent epidemiological studies have revealed that aging, physical and psychological stress, chronic pain, several metabolic disorders such as insulin resistance and established diabetes, alcoholism, inflammatory conditions, and vascular disorders such as arterial hypertension all may be associated with depression. The present review examines some of these depression-associated factors and the mechanisms by which they might give rise to vascular disorders such as atherosclerosis, microcirculation endothelial dysfunction, and interstitial disturbances leading to organ damage. A number of disorders involving the circulation can lead progressively and insidiously to large artery rigidity, remodeling of peripheral arteries, and alterations of the microcirculation of large blood vessels. Perturbations in vasa vasorum blood flow may contribute to atherogenesis, in addition to the influence of numerous cellular events involved in inflammation (tumor necrosis factor alpha, interleukin 1 beta, etc). Since Hans Selye first described the neuroendocrine cascade generated by experimentally induced stress half a century ago, phenomena such as the axonal release of neurotransmitters (including serotonin), accumulation of metabolites such as homocysteine, platelet-activating factor, and nitric oxide also have been implicated in the pathogenesis of depression. Moreover, vascular consequences of depression such as heart rate and pulse pressure variations may lead to endothelial dysfunction in critical microcirculation networks (cerebral, myocardial, and renal) and initiate physicochemical alterations in interstitial compartments adjacent to vital organs. The appropriate use of ambulatory monitoring of vascular parameters, such as heart rate and pulse pressure, and eventually, early identification of genetic and metabolic markers may prove helpful in the early detection of events preceding and predicting the clinical manifestations of depression. PMID- 15877314 TI - Depression in women. AB - Depression is the leading cause of disease-related disability in women. Epidemiological studies have shown that the lifetime prevalence of a major depressive disorder in women (21.3%) is almost twice that in men (12.7%). This ratio has been documented in different countries and ethnic groups. Sex differences relating to depression vary with age, with male and female children showing similar incidence rates. National comorbidity data reveal that sex differences in prevalence first appear around the age of 10 years and persist until midlife, after which they disappear. Therefore, women have the greatest risk for developing depressive disorders during their child-bearing years. Several biological processes are thought to be involved in the predisposition of women to depression, including genetically determined vulnerability, hormonal fluctuations related to various aspects of reproductive function, and an undue sensitivity to such hormonal fluctuations in brain systems that mediate depressive states. Psychosocial events such as role-stress, victimization, sex specific socialization, internalization coping style, and disadvantaged social status have all been considered to be contributors to the increased vulnerability of women to depression. Women are more susceptible than men to stress-induced depression and to changes in photoperiod (more than 80% of individuals with seasonal affective disorder are women). Depression in women may develop during different phases of the reproductive cycle (premenstrual dysphoric disorder, depression during pregnancy, postpartum depressive conditions, and menopausal depression). Other reproductive events such as infertility, miscarriage, oral contraceptives, and hormone replacement treatment have been reported to cause depression in women. PMID- 15877315 TI - Medical and psychiatric illness: different but concurrent! PMID- 15877316 TI - Report from the 12th Retrovirus Conference. Immune-based therapies for established HIV infection. PMID- 15877317 TI - Bibliography. Current world literature. Melanoma and other skin neoplasms. PMID- 15877318 TI - Myocardial stunning due to sudden emotional stress. PMID- 15877319 TI - Myocardial stunning due to sudden emotional stress. PMID- 15877320 TI - Vascular risk factors and diabetic neuropathy. PMID- 15877321 TI - Vascular risk factors and diabetic neuropathy. PMID- 15877322 TI - Health care in the 21st century. PMID- 15877323 TI - Health care in the 21st century. PMID- 15877324 TI - Health care in the 21st century. PMID- 15877325 TI - Parecoxib: renal failure. PMID- 15877326 TI - Valdecoxib: cutaneous and vascular risks. PMID- 15877327 TI - Endocrine-related resources from the National Institutes of Health. PMID- 15877329 TI - In vitro cell response to a polymer surface micropatterned by laser interference lithography. AB - This presentation will introduce laser interference lithography to prepare a periodic line and point micropatterns for study of cell-surface interactions. This process provides a straightforward micropatterning technique based on selective laser ablation of polymers utilizing the periodic energy distribution of two or more beam interference patterns. The micropatterns were characterized by atomic force microscopy, while the surface chemical modification was analyzed using X-ray photoelectron spectroscopy. Human pulmonary fibroblasts cultured on the surface of polycarbonate bearing line micropatterns were elongated, spindlelike, and oriented themselves along the line patterns with all different groove widths. In contrast, cells cultured on point patterns were also bipolar but showed no orientation. Further investigations demonstrated that human pulmonary fibroblast cells cultured on line and point micropatterns showed inflammatory response. PMID- 15877330 TI - Smart hydrogels for in situ generated implants. AB - The objective of this study was to explore the use of reverse thermo-responsive (RTG) polymers for generating implants at their site of performance, following minimally invasive surgical procedures. Aiming at combining syringability and enhanced mechanical properties, a new family of injectable RTG-displaying polymers that exhibit improved mechanical properties was created, following two different strategies: (1) to synthesize high-molecular-weight polymers by covalenty joining poly(ethylene glycol) and poly(propylene glycol) chains using phosgene as the coupling molecule and (2) to cross-link poly(ethylene oxide) (PEO)-poly(propylene oxide) (PPO)-PEO triblocks after end-capping them with triethoxysilane or methacrylate reactive groups. While the methacrylates cross linked rapidly, the triethoxysilane groups enabled the system to cross-link gradually over time. The chain-extended PEO/PPO copolymers had molecular weights in the 39 000-54 000 interval and exhibited improved mechanical properties. Reverse thermo-responsive systems displaying gradually increasing mechanical properties were generated by cross-linking triethoxysilane-capped (EO)(99) (PO)(67)-(EO)(99) (F127) triblocks. Over time, the ethoxysilane groups hydrolyzed and created silanol moieties that subsequently condensated. With the aim of further improving their mechanical behavior, F127 triblocks were reacted with methacryloyl chloride and the resulting dimethacrylate was subsequently cross linked in an aqueous solution at 37 degrees C. The effect of the concentration of the F127 dimethacrylate on the mechanical properties and the porous structure of the cross-linked matrixes produced was assessed. Rheometric studies revealed that the cross-linked hydrogels attained remarkable mechanical properties and allowed the engineering of robust macroscopic constructs, such as large tubular structures. The microporosity of the matrixes produced was studied by scanning electron microscopy. Monolayered conduits as well as structures comprising two and three layers were engineered in vitro, and their compliance and burst strength were determined. PMID- 15877331 TI - In situ structure and activity studies of an enzyme adsorbed on spectroscopically undetectable particles. AB - The structural characteristics and the activity of a hyperthermophilic endoglucanase were investigated upon adsorption. Silica (hydrophilic) and Teflon (hydrophobic) surfaces were selected for the study. The materials were specially designed so that the interaction of the particles with light was negligible, and the enzyme conformation in the adsorbed state was monitored in situ. The adsorption isotherms were determined, and the adsorbed endoglucanase was studied using a number of spectroscopic techniques, enzymatic activity tests, and dynamic light scattering. Experiments were performed at pH values below, at, and above the isoelectric point of the enzyme. It was shown that the enzyme adsorbed on the hydrophobic surface of Teflon with higher affinity as compared to the hydrophilic silica nanoparticles. In all cases, adsorption was followed by (slight) changes in the secondary structure resulting in decreased beta-structural content. The changes were more profound upon adsorption on Teflon. The adsorbed enzyme remained active in the adsorbed state in spite of the structural changes induced when interacting with the surfaces. PMID- 15877332 TI - Nanoparticulate system for efficient gene transfer into refractory cell targets. AB - A biocompatible, nanoparticulate formulation has been designed to retain, protect, and deliver adenoviral gene constructs over an extended time course. Such devices can be administered locally or systemically with low toxicity. A multipolymeric nanoparticulate system, featuring very high stability in physiologic media, was designed to allow efficient in vitro gene transfer. The efficacy of nanoparticulate delivery is effective in cell systems that are normally refractory to gene transfer, such as pancreatic islets and antigen presenting cells. The findings suggest a nonspecific uptake system that permits adenoviral particle release within the transfected cells. A comparison with literature data revealed that our system is efficient at much lower levels (at least three orders of magnitude) of infectious viral particles. PMID- 15877333 TI - Hydroxyapatite surface modified by L-lactic acid and its subsequent grafting polymerization of L-lactide. AB - A new method of surface modification of hydroxyapatite nanoparticles (n-HA) by surface grafting reaction of l-lactic acid and ring-opening polymerization of l lactide (LLA) was developed. Two modified HA nanoparticles were obtained: HA modified by l-lactic acid (l-HA) and HA grafting with poly(l-lactide) (PLLA; p HA). The modified surface of n-HA was attested by Fourier transformation infrared, (31)P MAS NMR, and thermal gravimetric analysis. The results showed that l-lactic acid could be easily grafted onto the n-HA surface by forming a Ca carboxylate bond and initiated by the hydroxyl group of the grafted l-lactic acid and LLA could be graft-polymerized onto the n-HA surface in the presence of stannous octanoate. The highest grafting amounts of l-lactic acid and PLLA were about 33 and 22 wt %, respectively. The modified HA/PLLA composites showed good mechanical properties and uniform microstructure. The tensile strength and modulus of the p-HA/PLLA composite containing 15 wt % of p-HA were 67 MPa and 2.1 GPa, respectively, while those of the n-HA/PLLA composites were 45 MPa and 1.7 GPa, respectively. The elongation at the break of the l-HA/PLLA composite containing 15 wt % l-HA could reach 44%, in comparison with 6.5% of the n-HA/PLLA composites containing 15 wt % n-HA. PMID- 15877334 TI - pH-triggered changes in assembling properties of beta-cyclodextrin-conjugated poly(epsilon-lysine) complexes. PMID- 15877335 TI - Engineering of biomaterials surfaces by hyaluronan. AB - This review addresses the area of study that defines the field of surface modification of biomedical materials and devices by hyaluronan (HA), as related to the exploitation of HA biological properties. To provide a comprehensive view of the subject matter, initial sections give a quick introduction to basic information on HA-protein and HA-cell interactions, together with some discussion on the bioactive role of HA in wound healing and related phenomena. This is followed by a description of current theories that correlate HA properties to its molecular structure in aqueous media, underlying how HA molecular details are crucial for its biological interaction and role. Finally, existing approaches to surface modification by HA are reviewed, stressing the need for HA-surface engineering founded on the knowledge and control of the surface-linked HA molecular conformation at the solid/aqueous interface. PMID- 15877336 TI - Phospholipid-stabilized Au-nanoparticles. AB - This communication outlines a simple two-step approach of modification of 1 nm diameter Au nanoparticles using an aqueous solution of (1,2-dipalmitoyl-sn glycero-3-phosphothio-ethanol) phospholipid (PL). Transmission electron microscopy as well as particle size analysis show that, as a result of PL reactions with Au particles, the initial Au nanoparticle size increases to 5 nm. Considering the size of the PL and their ability to form liposomes, 5 nm diameter spheres indicate that the PL bilayer was attached to the surface of Au particles and the PL-Au interactions are facilitated by the presence of thiol functionality. The change of surface electronic properties of PL-stabilized Au particles is manifested by the disappearance of the 217 and 290 nm absorbances due to 5d-6sp transitions in Au, which is likely attributed to the presence of S H functionalities which increase the free electron density of the particle. As a consequence, two surface plasmons resulting from a collective oscillation of electrons in response to UV excitation disappear. PMID- 15877337 TI - Recombinant protein-co-PEG networks as cell-adhesive and proteolytically degradable hydrogel matrixes. Part I: Development and physicochemical characteristics. AB - Toward the development of synthetic bioactive materials to support tissue repair, we present here the design, production, and characterization of genetically engineered protein polymers carrying specific key features of the natural extracellular matrix, as well as cross-linking with functionalized poly(ethylene glycol) (PEG) to form hybrid hydrogel networks. The repeating units of target recombinant protein polymers contain a cell-binding site for ligation of cell surface integrin receptors and substrates for plasmin and matrix metalloproteinases (MMPs), proteases implicated in wound healing and tissue regeneration. Hydrogels were formed under physiological conditions via Michael type conjugate addition of vinyl sulfone groups of end-functionalized PEG with thiols of cysteine residues, representing designed chemical cross-linking sites within recombinant proteins. Cross-linking kinetics was shown to increase with the pH of precursor solutions. The elastic moduli (G') and swelling ratios (Q(m)) of the resulting hydrogels could be varied as a function of the stoichiometry of the reacting groups and precursor concentration. Optima of G' and Q(m), maximum and minimum, respectively, were obtained at stoichiometry ratios r slightly in excess of 1 (r = cysteine/vinyl sulfone). The pool of technologies utilized here represents a promising approach for the development of artificial matrixes tailored for specific medical applications. PMID- 15877338 TI - Molecular dynamics simulations of aqueous pullulan oligomers. AB - Molecular dynamics (MD) simulations have been used to model small-angle X-ray scattering (SAXS) data on aqueous solutions of four oligomeric segments of the glucan pullulan: the trimer G(3) (comprising one polymer repeating unit), the hexamer (G(3))(2), the nonamer (G(3))(3), and the dodecamer (G(3))(4). The AMBER force field was used in conjunction with the GB/SA continuum solvation model to calculate both the mean global dimensions of the oligomers from the limiting small angle scattering behavior and the shorter range structural information implicit in the Debye scattering function at larger scattering angles. This same force field and solvation treatment were employed earlier by Liu et al. (Macromolecules 1999, 32, 8611-8620) with apparent success in a rotational isomeric state (RIS) treatment of the same experimental data. The present work discloses that, despite numerical success in modeling the SAXS data, the RIS treatment, which includes only the interactions within dimeric segments of the polymer chain, fails to account accurately for excluded volume effects at the range of 3-12 sugar residues in the polymer backbone. It is suggested that MD simulations using continuum solvation models can be used to circumvent errors inherent in the computationally efficient RIS treatments of polymer nano- and picosecond dynamics while at the same time avoiding the heavy computational requirements of all-atom methods. PMID- 15877339 TI - Site-specific binding of the 9.5 kilodalton DNA-binding protein ORF80 visualized by atomic force microscopy. AB - Atomic force microscopy (AFM) has been used to examine the binding properties of the DNA-binding protein ORF80 to DNA. ORF80 is a 9.5 kDa protein that binds site specifically to double-stranded DNA of the sequence TTAA-N(7)-TTAA. Direct sizing of the protein complexes on DNA fragments from the plasmid pRN1 with AFM shows that the protein ORF80 binds preferentially to two positions. These positions agree well with the ORF80 binding sites determined by footprinting analysis. The measurements allow an estimate of the stoichiometry of the DNA-protein complexes. In contrast to previous results, the single-molecule experiments suggest that only a low number of ORF80 molecules bind to a DNA-binding site. PMID- 15877340 TI - SEC-MALS-QELS study on the molecular conformation of cellulose in LiCl/amide solutions. AB - The SEC-MALS-QELS (size-exclusion chromatography equipped with multiangle light scattering and quasi-elastic light scattering detectors) method using lithium chloride/N,N-dimethylacetamide (LiCl/DMAc) and LiCl/1,3-dimethyl-2 imidazolidinone (LiCl/DMI) as mobile phases was applied to cellulose and cellulose tricarbanilate (CTC) samples with various average degree of polymerization (DP) values. Molecular conformations of cellulose and CTC in the solvents were then discussed and compared on the basis of the relationships between the radii of gyration (R(g,z) or S(2)(z)(1/2)), the hydrodynamic radii (R(h,z)), and weight-average DP (DP(w)) or the contour lengths (L(w)). The Benoit Doty theory for wormlike polymer chains was applied to the R(g) vs L(w) data obtained, and the theoretical curves with Kuhn segment lengths l(K) of around 18 nm were found to fit the data of both cellulose and CTC molecules in the solvents. It was concluded from the obtained results that both cellulose and CTC molecules have conformations essentially identical to each other in the solvents; they behave as typical semiflexible chains in good solvents. PMID- 15877341 TI - Molecular level investigations of the inter- and intramolecular interactions of pH-responsive artificial triblock proteins. AB - Intelligent materials that can undergo physical gelation in response to environmental stimuli have potential impacts in the bioengineering and biomedical fields where the entrapment of cellular or molecular species is desired. Here, we utilize atomic force microscopy (AFM) to perform molecular level investigations of designer artificial proteins that undergo physical gelation. These are engineered as triblock copolymers with independent interchain binding and solvent retention functions, namely, two terminal leucine zipper-like peptide sequences and a central alanylglycine rich sequence, respectively. AFM force measurements between probes and surfaces functionalized with molecules of this triblock protein revealed adhesive interactions that increased in average force and frequency as the pH was lowered from pH 11.2 to 7.4 to 4.5, reflecting an increase in the numbers of interacting molecular strands. In bulk solution, lowering the pH results in a viscous liquid to gel transition. The modular design of the triblock protein was also exploited for single molecule force spectroscopy investigations, which revealed altered intramolecular interactions in response to changes in pH. An increased understanding of the inter- and intramolecular forces involved in biomolecule driven gelation processes is not only of great fundamental interest in the study of the biomolecular systems involved but may also prove key in enabling the rational design of new generations of intelligent hydrogel systems. PMID- 15877342 TI - Fractalkine targeting with a receptor-mimicking peptide-amphiphile. AB - In this study we have designed the NTFR peptide-amphiphile that mimics a fragment of the N-terminus of the fractalkine receptor (CX(3)CR1) and specifically targets fractalkine, a novel adhesion molecule expressed on the surface of inflamed endothelial cells. Bioartificial membranes were constructed from mixtures of NTFR peptide-amphiphiles and DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) phospholipids, and the affinity and specificity of fractalkine for the synthetic NTFR was investigated with an atomic force microscope (AFM). Fractalkine was immobilized onto the AFM tips, and forces were collected between fractalkine and the bioartificial membranes. The adhesive interactions were studied at the collective level, when each adhesion event corresponded to the rupture of multiple biomolecular bonds. Retraction force profiles for the fractalkine-NTFR system exhibited single or multiple peaks and a small percentage of the force curves demonstrated stretching of the fractalkine-NTFR complex. Strong adhesion was measured when both DPPC and NTFR were present, compared to pure NTFR surfaces. This may be due to the fact that the DPPC molecule is shorter, and thus it can provide more space for the peptide headgroup to bend and expose its sequence at the interface. Specificity was demonstrated by comparing the NTFR fractalkine adhesion to the forces between the alpha(5)beta(1) integrin (an adhesion receptor expressed on the surface of endothelial cells) and other surfaces such as GRGDSP (the specific ligand for alpha(5)beta(1)), GRGESP (an inactive sequence), and NTFR. PMID- 15877343 TI - Preparation of magnetic microspheres from water-in-oil emulsion stabilized by block copolymer dispersant. AB - A new method for the preparation of magnetic microspheres is reported. The preparation involved first the dispersion of an aqueous phase, containing magnetite nanoparticles and a water-soluble homopolymer, into droplets in an organic medium using an amphiphilic block copolymer as the dispersant. This was followed by water distillation at a raised temperature from the aqueous droplets to yield polymer/magnetite particles. The structure of the particles was then locked in by a reagent being added to cross-link the water-soluble copolymer block and homopolymer. Since the hydrophobic block of the copolymer consisted of a protected polyester, the removal of the protective moieties from the coronal chains yielded poly(acrylic acid) or other functional polymers to render water dispersibility to the spheres and to enable biomolecule immobilization. PMID- 15877344 TI - CaCO3 biomineralization: acidic 8-kDa proteins isolated from aragonitic abalone shell nacre can specifically modify calcite crystal morphology. AB - Acidic proteins from many biogenic minerals are implicated in directing the formation of crystal polymorphs and morphologies. We characterize the first extremely acidic proteins purified from biomineralized aragonite. These abalone nacre proteins are two variants of 8.7 and 7.8 kDa designated AP8 (for aragonite proteins of approximately 8 kDa). The AP8 proteins have compositions dominated by Asx ( approximately 35 mol %) and Gly ( approximately 40 mol %) residues, suggesting that their structures have high Ca(2+)-binding capacity and backbone flexibility. The growth of asymmetrically rounded CaCO(3) crystals in the presence of AP8 reveals that both proteins preferentially interact with specific locations on the crystal surface. In contrast, CaCO(3) crystals grown with nacre proteins depleted of AP8 retain the morphology of unmodified calcite rhombohedra. Our observations thus identify sites of protein-mineral interaction and provide evidence to support the long-standing theory that acidic proteins are more effective crystal-modulators than other proteins from the same biomineralized material. PMID- 15877345 TI - Conformational study and hydrogen bonds detection on elastin-related polypeptides using X-ray photoelectron spectroscopy. AB - The chemical bonds of the pentapeptide sequence of elastin ValGlyGlyValGly (VGGVG), both in its monomer and polymer forms, were correlated with their XPS spectra through a well-established curve-fitting procedure. To aid in this correlation, the C1s, O1s, and N1s chemical shifts of the Boc-VGGVG-OEt, were validated by theoretical calculations, performed in the framework of the Koopman approximation of HF/6-31G molecular orbitals, leading to the "preferred" conformation of the protected monomer. Then the same curve-fitting procedure was adopted for interpreting the XPS spectra of the polypentapeptide as a powder, and the XPS results obtained both for monomer and polymer compounds were compared with those obtained by FT-IR. The polymer was then analyzed after deposition onto a silicon substrate, Si(100), either from methanol or water suspensions and the presence of hydrogen bonds was detected at the polymer/substrate interface and between the polymer chains. The "surface rearrangement" that could be inferred from XPS results strongly confirms that derived from AFM images previously obtained under the same experimental conditions. In particular, the observed amyloid conformation is stabilized by hydrogen bonds to water molecules included in the structure while the formation of the beaded string structure observed in deposits from methanolic suspension is probably mediated by hydrogen bonds to the hydrated silicon surface. PMID- 15877346 TI - Solution self-assembly of hybrid block copolymers containing poly(ethylene glycol) and amphiphilic beta-strand peptide sequences. AB - The self-assembly in aqueous solution of hybrid block copolymers consisting of amphiphilic beta-strand peptide sequences flanked by one or two PEG chains was investigated by means of circular dichroism spectroscopy, small-angle X-ray scattering, and transmission electron microscopy. In comparison with the native peptide sequence, it was found that the peptide secondary structure was stabilized against pH variation in the di- and tri-block copolymers with PEG. Small-angle X-ray scattering indicated the presence of fibrillar structures, the dimensions of which are comparable to the estimated width of a beta-strand (with terminal PEG chains in the case of the copolymers). Transmission electron microscopy on selectively stained and dried specimens shows directly the presence of fibrils. It is proposed that these fibrils result from the hierarchical self assembly of peptide beta-strands into helical tapes, which then stack into fibrils. PMID- 15877347 TI - Repeated rapid shear-responsiveness of peptide hydrogels with tunable shear modulus. AB - A pair of mutually attractive but self-repulsive decapeptides, with alternating charged/neutral amino acid sequence patterns, was found to co-assemble into a viscoelastic material upon mixing at a low total peptide concentration of 0.25 wt %. Circular dichroism spectroscopy of individual decapeptide solutions revealed their random coil conformation. Transmission electron microscopy images showed the nanofibrillar network structure of the hydrogel. Dynamic rheological characterization revealed its high elasticity and shear-thinning nature. Furthermore, the co-assembled hydrogel was capable of rapid recoveries from repeated shear-induced breakdowns, a property desirable for designing injectable biomaterials for controlled drug delivery and tissue engineering applications. A systematic variation of the neutral amino acids in the sequence revealed some of the design principles for this class of biomaterials. First, viscoelastic properties of the hydrogels can be tuned through adjusting the hydrophobicity of the neutral amino acids. Second, the beta-sheet propensity of the neutral amino acid residue in the peptides is critical for hydrogelation. PMID- 15877348 TI - Chain-chain interactions for methyl polygalacturonate: models for high methyl esterified pectin junction zones. AB - The ability of pectins to form gels in the presence of calcium is well-known, and it implies the interaction of carboxylate groups and bivalent ions. However, even when most of the galacturonic units are methyl esterified, pectins are able to form gels but only under certain experimental conditions. In this case, hydrogen bonding and hydrophobic interactions are believed to be responsible for gel formation, and it is likely, as in the other mechanisms of polysaccharide gel formation, that stable junction zones consist of cooperatively ordered chains linked together throughout nonbonded interactions to provide a three-dimensional network. To investigate the junction zones in HM-pectin gels, we investigated, by molecular modeling, all of the ways to associate two, and then three, fully methyl-esterified galacturonic acid chains. Two models are obtained: the first one is based on a packing of parallel chains; it agrees with the hypothetical model derived from fiber diffraction study; the second one displays an antiparallel orientation of the chains; it presents a better arrangement of the chains and, theoretically, a much lower potential energy. In both cases, all of the favorable associations occur within a network of hydrogen bonds and of hydrophobic contacts. PMID- 15877349 TI - Viscoelastic properties and dynamics of porcine gastric mucin. AB - Gastric mucin is a glycoprotein known to undergo a pH-dependent sol-gel transition that is crucial to the protective function of the gastric mucus layer in mammalian stomachs. We present microscope-based dynamic light scattering data on porcine gastric mucin at pH 6 (solution) and pH 2 (gel) with and without the presence of tracer particles. The data provide a measurement of the microscale viscosity and the shear elastic modulus as well as an estimate of the mesh size of the gel formed at pH 2. We observe that the microscale viscosity in the gel is about 100-fold lower than its macroscopic viscosity, suggesting that large pores open up in the gel reducing frictional effects. The data presented here help to characterize physiologically relevant viscoelastic properties of an important biological macromolecule and may also serve to shed light on diffusive motion of small particles in the complex heterogeneous environment of a polymer gel network. PMID- 15877350 TI - Application of catalytic ozone chemistry for improving biodiesel product performance. AB - Ozonolysis of methyl soyate (biodiesel) was conducted in the presence of methanol, dichloromethane (solvent), and triethylamine (catalyst) at -75 degrees C. Structural analysis, including FTIR, GC, and GC-MS, showed that the total amount of double bonds in the mixture was reduced by more than 90% after 2 h of ozonolysis. All of the esters predicted by this novel application of ozone reaction chemistry were successfully produced. Other major components were identified by GC-MS. Thermogravimetric analysis showed a dramatic decrease in the onset volatilization temperature from 135 to 73 degrees C, making ozonated biodiesel fuel comparable to diesel fuel (76 degrees C). Differential scanning calorimetric studies showed that the cooling curves for both methyl soyate and ozonated methyl soyate displayed two exothermic regions. The onset freezing temperature of ozonated methyl soyate in the "colder" region was significantly reduced from -63 to -86 degrees C. Furthermore, the degree of crystallinity in the "hotter" region was also reduced. PMID- 15877351 TI - Copolymerizations of epsilon-caprolactone and glycolide-a comparison of tin(II)octanoate and bismuth(III)subsalicylate as initiators. AB - Copolymerizations of epsilon-caprolactone (epsilonCL) and glycolide (GL) were conducted in bulk at 120 degrees C with variation of the reaction time. Either Sn(II) 2-ethylhexanoate (SnOct(2)) or bismuth(III)subsalicylate (BiSS) were used as initiators combined with tetra(ethylene glycol) as co-initiator. The resulting copolyesters were analyzed by (1)H and (13)C NMR spectroscopy with regard to the total molar composition and to the sequence of the comonomers. Furthermore, two series of copolymerizations (either Sn- or Bi-initiated) were performed at constant time with variation of the temperature. It was found that BiSS favors alternating sequences more than SnOct(2). Time-conversion curves and MALDI-TOF mass spectrometry of homopolymerization suggest that SnOct(2) is the more efficient transesterification catalyst. A hypothetical reaction mechanism is discussed. PMID- 15877352 TI - Build-up of collagen and hyaluronic acid polyelectrolyte multilayers. AB - We have used a novel polyelectrolyte multilayer (PEM) coating consisting of the polyelectrolytes collagen and hyaluronic acid. The build-up by the layer-by-layer deposition technique is outlined by ex situ and in situ ellipsometric measurements. When collagen was added, the thickness of the PEM was increased, and the refractive index was decreased. Corresponding but opposite effects were noted when hyaluronic acid was added. These changes are considered to be explained by a diffusion mechanism. It was also found that the PEM layers were unstable at physiological pH. However, by cross-linking using N-(3-di methylaminopropyl)-N'-ethylcarbodiimide together with N-hydroxysuccinimide, a stable PEM layer resulted. These tissue friendly PEM layers are expected to have a great impact in the design of artificial extracellular matrixes. Also, the insertion of fluorescence labels demonstrates the potential for incorporation of other functionalities. PMID- 15877353 TI - Synthesis of conducting polyelectrolyte complexes of polyaniline and poly(2 acrylamido-3-methyl-1-propanesulfonic acid) catalyzed by pH-stable palm tree peroxidase. AB - Comparison of the stability of five plant peroxidases (horseradish, royal palm tree leaf, soybean, and cationic and anionic peanut peroxidases) was carried out under acidic conditions favorable for synthesis of polyelectrolyte complexes of polyaniline (PANI). It demonstrates that palm tree peroxidase has the highest stability. Using this peroxidase as a catalyst, the enzymatic synthesis of polyelectrolyte complexes of PANI and poly(2-acrylamido-3-methyl-1 propanesulfonic acid) (PAMPS) was developed. The template polymerization of aniline was carried out in aqueous buffer at pH 2.8. Varying the concentrations of aniline, PAMPS, and hydrogen peroxide as reagents, favorable conditions for production of PANI were determined. UV-vis-NIR absorption and EPR demonstrated that PAMPS and PANI formed the electroactive complex similar to PANI doped traditionally using low molecular weight sulfonic acids. The effect of pH on conformational variability of the complex was evaluated by UV-vis spectroscopy. Atomic force microscopy showed that a size of the particles of the PANI-PAMPS complexes varied between 10 and 25 nm, depending on a concentration of PAMPS in the complex. The dc conductivity of the complexes depends also on the content of PAMPS, the higher conductivity being for the complexes containing the lower content of the polymeric template. PMID- 15877354 TI - Investigations on the solubility behavior of cyanophycin. Solubility of cyanophycin in solutions of simple inorganic salts. AB - On the basis of a previous report on the occurrence of water-soluble cyanophycin (CGP, cyanophycin granule polypeptide) in a recombinant strain of Escherichia coli expressing the cyanophycin synthetase (CphA) of Desulfitobacterium hafniense published by others, the conditions of its production were investigated in this study. Although the incubation temperature, aeration level, and NaCl concentration during cultivation had effects on the in vivo production of water soluble CGP, it could be isolated as a major variant irrespective of the cultivation conditions. The occurrence of the soluble variant was also not dependent on the E. coli host or on the origin of cphA. Furthermore, it was shown that water-insoluble CGP can be in vitro solubilized to extents of up to about 80% (w/w) in solutions of different inorganic salts such as LiCl, NaCl, KCl, RbCl, KBr, MgCl(2), or CaCl(2). Evidence was obtained that the salt ions bind tightly to CGP. If the ions were not removed from the salt solution by dialysis or dilution, the CGP remained stable in solution. This method to solubilize water insoluble CGP could also be applied to high concentrations of the polymer. CGP that remained insoluble after the first treatment could only marginally be solubilized in following treatments. The polydisperse CGP molecules were solubilized to the same extent over the whole molecular weight range with no preference of a particular molecular weight. PMID- 15877355 TI - Polyphosphazene polyelectrolytes: a link between the formation of noncovalent complexes with antigenic proteins and immunostimulating activity. AB - Polyphosphazene polyelectrolytes are potent immunostimulants. Their in vivo performance has been demonstrated for various antigens in a number of animal models. To improve understanding of the mechanism of action, we performed a comparative study in a model system: bovine serum albumin, BSA poly[di(carboxylatophenoxy)phosphazene], PCPP, in vitro and in vivo. Multi-angle laser light scattering (MALLS) and size-exclusion HPLC methods were used to investigate polyphosphazene-protein formulations in an attempt to establish correlations between their physicochemical behavior and immunostimulating activity. These studies revealed the formation of water-soluble noncovalent protein-polymer complexes in the system. It was shown that both the amount of bound protein and the complex conformation could play an important role in the in vivo performance of the polyphosphazene polyelectrolytes. PMID- 15877356 TI - New route of deacetylation of alpha- and beta-chitins by means of freeze--pump out--thaw cycles. AB - 1st and 2nd heterogeneous deacetylations of alpha- and beta-chitins in a multistep process by means of freeze-pump out-thaw (FPT) cycles in the presence of 50% (w/v) NaOH, for temperatures ranging from 80 to 110 degrees C, were compared to the classical method using argon as the medium atmosphere. It was clearly demonstrated that FPT cycles extensively improved the reaction effectiveness by opening the crystalline structure of the two chitins and made them more permeable to alkaline solutions. The acetylated groups being more accessible, the preexponential factors of the reaction consequently enhanced, whereas the activation energies slightly increased, attesting to a more sensitive reaction to temperature. Compared to the usual method that mainly led to block copolymers for intermediate DAs, the statistical copolymers proceeded by FPT cycles improved the solubility of the samples and strongly modified the reaction mechanisms. For the very first time, the quasi-full absence of dioxygen in the reaction medium allowed us to estimate the non-oxidative degradation of the copolymer in alkaline conditions, independently of the oxydo-reductive process. Thus, heterogeneous deacetylation by means of FPT cycles exhibited all of the advantages of a reaction in homogeneous conditions (high solubility of the samples produced, reaction independent of the starting crystalline structure) without the drawbacks (low efficiency, severe degradation). It allowed us to generate fully deacetylated chitosans with the highest molecular weights never published in the literature. PMID- 15877357 TI - Transport properties of EVAl-starch-alpha amylase membranes. AB - We investigated the influence of various physicochemical parameters on the morphology and time-porosity formation of membranes composed of ethylene-vinyl alcohol, starch, and alpha-amylase. In particular, we determined that (1) it is possible to obtain a membrane with desired porosity by phase inversion in an appropriate water-ethanol mixture and (2) the enzymatic bioerosion is controlled by the amount of alpha-amylase present in the blend. Although no experiments involving drugs were carried out, the delivery properties of the film were determined by measuring the Darcy permeability, the effective diffusivity, and the mean reaction rate of the membranes, relating them to the modality of membrane preparation, the amount of enzyme present within the membrane, and the incubation time of the samples in a buffer solution. Simple theoretical models of the delivery properties of the membranes were developed, leading to predictions that were in good agreement with the experimental results. PMID- 15877358 TI - 13C Nuclear magnetic relaxation study of segmental dynamics of hyaluronan in aqueous solutions. AB - (13)C spin-lattice relaxation times (T(1)) and nuclear Overhauser enhancements (NOE) were measured as a function of temperature and magnetic field strength for the hetero-polysaccharide hyaluronan in water solutions. The relaxation data of the endocyclic ring carbons were successfully interpreted in terms of chain segmental motions by using the bimodal time-correlation function of Dejean de la Batie, Laupretre and Monnerie. On the basis of the calculated correlation times for segmental motion and amplitudes of librational motions of the C-H vectors at the various carbon sites of the HA repeating unit, we concluded that intramolecular hydrogen bonding of the secondary structure of HA plays a major role in the conformational flexibility of this carbohydrate molecule. The internal rotation of the free hydroxymethyl groups about the exocyclic C-5-C-6 bonds superimposed on segmental motion has been described as a diffusion process of restricted amplitude. The rate and amplitude of the internal rotation indicate that the hydroxymethyl groups are not involved in intramolecular hydrogen bonding. Finally, the motional parameters describing the local dynamics of the HA chain were correlated with the secondary structure of HA in aqueous solutions. PMID- 15877359 TI - Effects of electrospinning and solution casting protocols on the secondary structure of a genetically engineered dragline spider silk analogue investigated via Fourier transform Raman spectroscopy. AB - Micrometer and submicrometer diameter fibers of recombinant dragline spider silk analogues, synthesized via protein engineering strategies, have been electrospun from 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) and compared with cast films via Raman spectroscopy in order to assess changes in protein conformation that may result from the electrospinning process. Although the solvent casting process was shown to result in predominantly beta-sheet conformation similar to that observed in the bulk, the electrospinning process causes a major change in conformation from beta-sheet to alpha-helix. A possible mechanism involving electric field induced stabilization of alpha-helical segments in HFIP solution during the electrospinning process is discussed. PMID- 15877360 TI - Preparation and evaluation of trimethylsilylated chitin as a versatile precursor for facile chemical modifications. AB - Trimethylsilylation of chitin was studied in detail to establish a reliable method, and the properties of the resulting product were elucidated. Chitin was successfully trimethylsilylated with a mixture of hexamethyldisilazane and trimethylsilyl chloride in pyridine. Compared to alpha-chitin, beta-chitin was much more reactive and advantageous as a starting material to prepare fully substituted chitin in a simple manner, though alpha-chitin also underwent full silylation under appropriate conditions. The resulting silylated chitin was characterized by marked solubility in common organic solvents and by easy desilylation to regenerate hydroxy groups, which enabled clean preparation of chitin films. The reactivity of the silylated chitin was examined by treating with triphenylmethyl chloride and acetic anhydride as typical alkylating and acylating reagents, and complete substitutions were readily accomplished. The silylated chitin has thus proved to be a superb precursor for modification reactions. PMID- 15877361 TI - Swelling behavior of hyaluronic acid/polyallylamine hydrochloride multilayer films. AB - The reversible swelling behavior of multilayer films containing hyaluronic acid and polyallylamine hydrochloride was investigated using in situ ellipsometry, since many of the natural functions and applied uses of hyaluronic acid are related to the extraordinary ability of this biopolymer to swell, and to respond conformationally to the local solution environment. This swelling was observed to be substantial, and depended strongly on the film thickness, the pH conditions used to prepare the films, and the swelling solution pH and ionic strength. The swelling results were also rationalized in terms of the dissociation behavior of the polyelectrolytes in the multilayer assemblies, measured by the zeta potential, on colloidal particles. The films were found to swell by as much as 8 times their dry thickness, and the extent of film hydration was observed to depend on the thickness of the films in a nonlinear fashion. This was related to the internal structure of the films, which is dictated by the assembly pH conditions. In addition, the swelling solution pH and ionic strength influence the electrostatic environment in the films and, in turn, have a substantial effect on the overall swelling behavior. PMID- 15877362 TI - Purification and characterization of a vaterite-inducing peptide, pelovaterin, from the eggshells of Pelodiscus sinensis (Chinese soft-shelled turtle). AB - Proteins play a crucial role in the biomineralization of hard tissues such as eggshells. We report here the purification, characterization, and in vitro mineralization studies of a peptide, pelovaterin, extracted from eggshells of a soft-shelled turtle. It is a glycine-rich peptide with 42 amino acid residues and three disulfide bonds. When tested in vitro, the peptide induced the formation of a metastable vaterite phase. The floret-shaped morphology formed at a lower concentration ( approximately 1 microM) was transformed into spherical particles at higher concentrations (>500 microM). The solution properties of the peptide are investigated by circular dichroism (CD), fluorescence emission spectroscopy, and dynamic light scattering (DLS) experiments. The conformation of pelovaterin changed from an unordered state at a low concentration to a beta-sheet structure at high concentrations. Fluorescence emission studies indicated that the quantum yield is significantly decreased at higher concentrations, accompanied by a blue shift in the emission maximum. At higher concentrations a red-edge excitation shift was observed, indicating the restricted mobility of the peptide. On the basis of these observations, we discuss the presence of a peptide concentration dependent monomer-multimer equilibrium in solution and its role in controlling the nucleation, growth, and morphology of CaCO(3) crystals. This is the first peptide known to induce the nucleation and stabilization of the vaterite phase in solution. PMID- 15877363 TI - Size control of styrene oxide-ethylene oxide diblock copolymer aggregates with classical surfactants: DLS, TEM, and ITC study. AB - The interactions between the diblock copolymer S(15)E(63) and the surfactants sodium dodecyl sulfate (SDS), sodium decyl sulfate (SDeS), and sodium octyl sulfate (SOS) have been investigated by dynamic light scattering (DLS), transmission electron microscopy (TEM), and isothermal titration calorimetry (ITC). The surfactants with the same headgroup differentiate in their chain length. At 20 degrees C, the block copolymer is associated into micelles with a hydrodynamic radius of 11.6 nm, which is composed of a hydrophobic styrene oxide (S) core and a water-swollen oxypolyethylene (PEO or E) corona. The different copolymer/surfactant systems have been studied at a constant copolymer concentration of 2.5 g dm(-3) and in a vast range of surfactant concentrations, from 7.5 x 10(-6) up to 0.75 M. When SDS and SDeS are added to the block copolymer solution, different regions are observed in the DLS data: at low surfactant concentrations (c < 1.0 x 10(-4) M), single surfactant molecules associate with the copolymer micelle, probably the former being solubilized in the micelle core, leading to a certain disruption of the mixed micelle due to repulsive electrostatic interactions between surfactant headgroups followed by a stabilization of the mixed micelle. At higher concentrations (1.0 x 10(-4) < c < 0.1 M), two types of copolymer-surfactant complexes coexist: one large copolymer rich/surfactant complex and one small complex consisting of one or a few copolymer chains and rich in surfactants. At higher SDS and SDeS concentrations, complete disintegration of mixed micelles takes place. In contrast, SOS S(15)E(63) interactions are less important up to surfactant concentrations of 0.05 M due to its higher hydrophilicity, reducing the hydrophobic interactions between surfactant alkyl chains and copolymer micelles. At concentration larger than the critical aggregation concentration (cac) of the system, 0.05 M, disruption of copolymer micelles occurs. These regions have been confirmed by transmission electron microscopy. On the other hand, the titration calorimetric data for SDS and SDeS present an endothermic increase indicating the formation of mixed copolymer-rich-surfactant micelles. From that point, important differences in the ITC plot for both surfactants are present. However, the ITC curve obtained after titration of a SOS solution in the copolymer solution is quite similar to that of its titration in water. PMID- 15877364 TI - Structural analysis of the capsular polysaccharide from Sinorhizobium fredii HWG35. AB - We have determined the structure of a capsular polysaccharide from Sinorhizobium fredii HWG35. This polysaccharide was isolated following the standard protocols applied for lipopolysaccharide isolation. On the basis of monosaccharide analysis, methylation analysis, mass spectrometric analysis, one-dimensional (1)H and (13)C NMR, and two-dimensional NMR experiments, the structure was shown to consist of a polymer having the following disaccharide repeating unit: -->6)-2,4 di-O-methyl-alpha-d-Galp-(1-->4)-beta-d-GlcpA-(1-->. Strain HWG35 produces a capsular polysaccharide that does not show the structural motif (sugar-Kdx) observed in those S. fredii strains that, while effective with Asiatic soybean cultivars, are unable to form nitrogen-fixing nodules with American soybean cultivars. Instead, the structure of the capsular polysaccharide of S. fredii HWG35 is in line with those produced by strains HH303 (rhamnose and galacturonic acid) and B33 (4-O-methylglucose-3-O-methylglucuronic acid), two S. fredii strains that form nitrogen-fixing nodules with both groups of soybean cultivars. Hence, in these three strains that effectively nodulate American soybean cultivars, the repeating unit of the capsular polysaccharide is composed of two hexoses, one neutral (methylgalactose, rhamnose, or methylglucose) and the other acidic (glucuronic, galacturonic, or methylglucuronic acid). PMID- 15877365 TI - Understanding structure-stability relationships of Candida antartica lipase B in ionic liquids. AB - Two different water-immiscible ionic liquids (ILs), 1-ethyl-3-methylimidizolium bis(trifluoromethylsulfonyl)imide and butyltrimethylammonium bis(trifluoromethylsulfonyl)imide, were used for butyl butyrate synthesis from vinyl butyrate catalyzed by Candida antarctica lipase B (CALB) at 2% (v/v) water content and 50 degrees C. Both the synthetic activity and stability of the enzyme in these ILs were enhanced as compared to those in hexane. Circular dichroism and intrinsic fluorescence spectroscopic techniques have been used over a period of 4 days to determine structural changes in the enzyme associated with differences in its stability for each assayed medium. CALB showed a loss in residual activity higher than 75% after 4 days of incubation in both water and hexane media at 50 degrees C, being related to great changes in both alpha-helix and beta-strand secondary structures. The stabilization of CALB, which was observed in the two ILs studied, was associated with both the maintenance of the 50% of initial alpha helix content and the enhancement of beta-strands. Furthermore, intrinsic fluorescence studies clearly showed how a classical enzyme unfolding was occurring with time in both water and hexane media. However, the structural changes associated with the incubation of the enzyme in both ILs might be attributed to a compact and active enzyme conformation, resulting in an enhancement of the stability in these nonaqueous environments. PMID- 15877366 TI - Multiphoton excited fabrication of collagen matrixes cross-linked by a modified benzophenone dimer: bioactivity and enzymatic degradation. AB - Multiphoton excited (MPE) photochemistry is used to fabricate model tissue engineering scaffolds directly from types I, II, and IV collagen. A modified benzophenone dimer (BPD) provides the photoactivation and becomes incorporated into the resulting collagen matrixes. Unlike xanthene photochemistries, the benzophenone dimer can be used in acidic environments, where most forms of collagen have the greatest solubility. The minimum feature sizes are investigated by using two- and three-photon excitation, where the latter provides for superior "resolution" and suggests that collagen structures can be fabricated on the size scales of focal contacts. The resulting structures display excellent retention of bioactivity as evidenced by highly specific cell adhesion as well as immunofluorescence labeling. Structural and chemical aspects of the collagen matrixes are probed through measuring the enzymatic degradation through specific and nonspecific proteases, as the resulting relative rates are consistent with the activity of these enzymes. The degradation rates can also be controlled through varying the cross-link density in the matrixes, which is achieved through tuning the exposure dose during the fabrication process. The degradation rates are also found to be consistent with swelling/shrinking measurements and thus the average mesh size of the matrixes. In all cases the enzymatic degradations are well-fit single exponentials, suggesting that the matrixes can be fabricated with a priori knowledge of their structural properties. These results coupled with the resulting bioactivity suggest that the multiphoton fabrication process may be a powerful tool for the creation of cell-sized tissue engineering scaffolds. PMID- 15877367 TI - Electrostatic adsorption of heme proteins alternated with polyamidoamine dendrimers for layer-by-layer assembly of electroactive films. AB - A novel thin film of heme proteins, including hemoglobin (Hb), myoglobin (Mb), and catalase (Cat), was successfully assembled layer by layer with polyamidoamine (PAMAM) dendrimers on different solid surfaces. At pH 7.0, protonated PAMAM possesses positive surface charges, whereas the proteins have net negative surface charges at pH above their isoelectric points. Thus, layer-by-layer {PAMAM/protein}(n)() films were assembled with alternate adsorption of oppositely charged PAMAM and proteins from their aqueous solutions mainly by electrostatic interaction. The assembly process was monitored by quartz crystal microbalance (QCM), UV-vis spectroscopy, and cyclic voltammetry (CV). The growth of the protein multilayer films was regular and linear, whereas the electroactivity of the films was only extended to a few bilayers. CVs of {PAMAM/protein}(n)() films showed a pair of well-defined and nearly reversible peaks characteristic of the protein heme Fe(III)/Fe(II) redox couples. Although {PAMAM/Hb}(n)() and {PAMAM/Mb}(n)() films showed very similar properties, {PAMAM/Cat}(n)() films displayed different and unique characters. The substrates with biological or environmental significance, such as oxygen, hydrogen peroxide, trichloroacetic acid, and nitrite, were catalytically reduced at {PAMAM/protein}(n)() film electrodes, showing the potential applicability of the films as new types of biosensors or bioreactors based on direct electrochemistry of the proteins. Both the electrochemical and electrocatalytic activity of {PAMAM/protein}(n)() films can be tailored precisely by controlling the number of bilayers or the film thickness. PMID- 15877368 TI - Poly(vinyl alcohol) nanofibers by electrospinning as a protein delivery system and the retardation of enzyme release by additional polymer coatings. AB - Protein-loaded (bovine serum albumin (BSA) or luciferase) poly(vinyl alcohol) (PVA) nanofibers were obtained by electrospinning. Poly(p-xylylene) (PPX, also coined as parylene) coated PVA/BSA nanofibers were prepared by chemical vapor deposition (CVD). The release of BSA from PVA nanofibers under physiological conditions was monitored by absorption spectroscopy. Burst release of BSA was noted with uncoated PVA nanofibers. In contrast, PPX-coated nanofibers exhibited a significantly retarded release of BSA depending on the coating thickness of PPX (ranging from 40 to 300 nm). Luciferase was used here as model enzyme, which after electrospinning retained its enzyme activity. This preservation of enzyme activity and the continuous release of the intact enzyme from the immersed fibers meets a fundamental prerequisite for the application of enzymes or other sensitive agents released from electrospun nanofibers under physiological conditions. PMID- 15877369 TI - Heat-resistant oxygen-carrying hemoproteins consist of recombinant xylanases and synthetic iron(II) porphyrin. AB - Synthetic iron(II) porphyrin (FeP) is equivalently incorporated into recombinant Thermotoga maritima xylanase B (TMX; family F/10 of glycoside hydrolase), producing a heat-resistant artificial hemoprotein (TMX-FeP) that can bind and release oxygen (O(2)) in aqueous medium (pH 7.3, 25 degrees C) in the same manner as hemoglobin and myoglobin. The oxygenated species was sufficiently stable; the half-lifetime against the ferric state (tau(1/2)) was 5 h. This O(2)-carrying hemoprotein showed a high degree of thermal stability over a wide range of temperatures up to 90 degrees C (tau(1/2) = 5 min at 90 degrees C and 9 min at 75 degrees C). Dictyoglomus thermophilum xylanase B (DTX; family G/11) also incorporates FeP, and DTX-FeP showed identical O(2)-binding parameters and thermostability. TMX-FeP is capable of catalyzing the beta-1,4-d-xylan hydrolysis reaction. Its larger K(m) value compared to that of TMX itself suggested competitive FeP binding to the active site of the host enzyme. PMID- 15877370 TI - Multilayer DNA/poly(allylamine hydrochloride) microcapsules: assembly and mechanical properties. AB - We report the preparation, characterization, and mechanical properties of DNA/poly(allylamine hydrochloride) (PAH) multilayer microcapsules. The DNA/PAH multilayers were first constructed on a planar support to examine their layer-by layer buildup. Surface plasmon resonance spectroscopy (SPR) showed a nonlinear growth of the assembly upon each bilayer deposited independently on a concentration of salt. A weak increase in the film thickness with the DNA concentration was, however, detected. A post-treatment of the multilayers in the salt solutions has shown a thinning of the film. The optimal conditions of the planar film growth were used to deposit the same multilayers on the surface of colloidal templates and to study their roughness and morphology with the atomic force microscope (AFM) imaging. When an outer layer is formed by DNA, we observe large domains of oriented parallel DNA loops, while an outer layer formed by PAH shows highly porous morphology. The dissolution of colloidal templates led to a formation of highly porous DNA/PAH microcapsules. We probe their mechanical properties by measuring force-deformation curves with the AFM-related setup. The experiment suggests that the DNA/PAH capsules are softer than capsules made from the flexible polyelectrolytes studied before. The softening is due to both higher permeability and smaller Young's modulus of the shell material. The Young's modulus of the DNA/PAH shells increases after post-treatment in salt solutions of relatively low concentration. PMID- 15877371 TI - Nitrocinnamate-functionalized gelatin: synthesis and "smart"hydrogel formation via photo-cross-linking. AB - Gelatin having p-nitrocinnamate pendant groups (Gel-NC) was prepared via an efficient one-pot synthesis, yield >87%. (1)H NMR data indicated that 1 mol of gelatin was modified with 18 +/- 6 mol of the photosensitive group. Upon exposure to low-intensity 365 nm UV light and in the absence of photoinitiators or catalysts, Gel-NC cross-linked within minutes into a gelatin-based hydrogel as monitored by UV-vis spectroscopy. The degree of swelling of this biodegradable hydrogel in aqueous solutions responded to changes in Gel-NC concentration levels, the ionic strength of the aqueous solutions, and photo-cross-linking time. Topography changes associated with phase transition resulting from "photocleavage" of the hydrogel network with 254 nm UV light were studied with AFM. Both Gel-NC and its hydrogel expressed low toxicity to human neonatal fibroblast cells. In addition, gelatin-based microgels were prepared via the photo-cross-linking of Gel-NC within inverse micelles. PMID- 15877372 TI - Determination of the number-average degree of polymerization of cellodextrins and cellulose with application to enzymatic hydrolysis. AB - A rapid and accurate method for determining the number-average degree of polymerization (DP(n)) was established for insoluble cellulose and soluble cellodextrins as the ratio of glucosyl monomer concentration determined by the phenol-sulfuric acid method divided by the reducing-end concentration determined by a modified 2,2'-bicinchoninate (BCA) method. The modified BCA method, featuring incubation at 75 degrees C for 30 min, did not result in beta glucosidic bond cleavage, whereas substantial cleavage was observed at higher temperature. Solubilization of insoluble cellulose in cold phosphoric acid prior to measurement of the reducing-end concentration by the BCA method was found not to be necessary for several model celluloses such as microcrystalline cellulose, but such solubilization was required for large fibers of cellulose such as Whatman No. 1 filter paper. The phenol-sulfuric acid method can be used for measuring the glucosyl monomer concentration of soluble cellodextrins, and also for insoluble cellulose if preceded by a liquefaction step. Standard deviations of < or =2% were obtained for both reducing and glucosyl monomer determination and of < or =3% for overall determination of DP. By use of the reported method, hydrolysis of phosphoric acid-swollen cellulose (PASC) by the Trichoderma reesei cellulase system was shown to result in a rapid decrease in DP as hydrolysis proceeded. By contrast, the DP of Avicel remained nearly constant during hydrolysis. The specific enzymatic cellulose hydrolysis rate is 100-fold higher for PASC as compared to Avicel. PMID- 15877373 TI - Expression and characterization of a four-alpha-helix bundle protein that binds the volatile general anesthetic halothane. AB - The structural features of volatile anesthetic binding sites on proteins are being investigated with the use of a defined model system consisting of a four alpha-helix bundle scaffold with a hydrophobic core. The current study describes the bacterial expression, purification, and initial characterization of the four alpha-helix bundle (Aalpha(2)-L1M/L38M)(2). The alpha-helical content and stability of the expressed protein are comparable to that of the chemically synthesized four-alpha-helix bundle (Aalpha(2)-L38M)(2) reported earlier. The affinity for binding halothane is somewhat improved with a K(d) = 120 +/- 20 microM as determined by W15 fluorescence quenching, attributed to the L1M substitution. Near-UV circular dichroism spectroscopy demonstrated that halothane binding changes the orientation of the aromatic residues in the four-alpha-helix bundle. Nuclear magnetic resonance experiments reveal that halothane binding results in narrowing of the peaks in the amide region of the one-dimensional proton spectrum, indicating that bound anesthetic limits protein dynamics. This expressed protein should prove to be amenable to nuclear magnetic resonance structural studies on the anesthetic complexes, because of its relatively small size (124 residues) and the high affinities for binding volatile anesthetics. Such studies will provide much needed insight into how volatile anesthetics interact with biological macromolecules and will provide guidelines regarding the general architecture of binding sites on central nervous system proteins. PMID- 15877374 TI - Mucous systems show a novel mechanical response to applied deformation. AB - Mucous secretions have a wide range of biological functions that are intimately linked with their rheological properties. In addition, many mucous secretions are exposed to significant stress and deformation during physiological function. This study has examined the rheological response of three mucous systems, native pig gastric mucus, purified mucin gels, and mucin alginate gels, to increasing applied stress to a level sufficient to induce flow behavior. A novel, frequency dependent stress hardening was observed in all three systems. This hardening behavior may play a significant role in the ability of mucous systems to resist mechanical disruption in the physiological state. PMID- 15877376 TI - Polysaccharide-polynucleotide complexes. Part 32. Structural analysis of the curdlan/poly(cytidylic acid) complex with semiempirical molecular orbital calculations. AB - Natural Curdlan adopts a right-handed 6(1) triple helix, in which the constituting glucan chains are underpinned with each other by the intermolecular hydrogen bonds. Curdlan can form a stoichiometric complex with polynucleotides [e.g., poly(cytidylic acid), poly(C)]. In this paper, we carried out the MOPAC (semiempirical molecular-orbital package) calculation to examine the molecular structure of the Curdlan/poly(C) complex. The calculation exhibited that two types of hydrogen bonds are formed between the Curdlan and the poly(C); the third nitrogen (N3) in cytosine forms a hydrogen bond with the second OH of one Curdlan chain, and the proton of N4 is interacting with the O2 of another Curdlan chain. In our model, the helix diameter of poly(C) is expanded from 11.0 to 15.3 A upon complexation. Despite such large conformational changes, the 6(1) helix structure of poly(C) was maintained even after the complexation. This fact is complementary to the experimental fact that the complexation does not change the band shape of the circular dichroism of poly(C). The chain length dependence of the reaction enthalpy indicated that the complexation becomes thermodynamically more favorable with the chain length increasing. This feature is also consistent with the experimental data. PMID- 15877375 TI - Conformational behavior of chemically reactive alanine-rich repetitive protein polymers. AB - The synthesis of protein-based polymers with controlled conformational properties and functional group placement offers many opportunities for the design of advanced materials. In this work, protein engineering methods have been used to produce repetitive alanine-rich protein polymers with the sequence [(AAAQ)(5)(AAAE)(AAAQ)(5)](x) (x = 2 and 6); these macromolecules may mimic architectural features of certain alanine-rich helical sequences found in natural proteins. Various proteins from this family can be readily expressed and purified from Escherichia coli. Circular dichroic spectroscopy (CD) characterization demonstrates that the purified proteins are highly helical under a variety of conditions. Thermal analysis of [(AAAQ)(5)(AAAE)(AAAQ)(5)](2) via differential scanning calorimetry (DSC) and CD indicates that the protein undergoes a reversible helix-coil transition at approximately 45 degrees C and that the protein conformation can be manipulated at elevated temperatures depending on solution conditions. The demonstrated conformational properties of these artificial proteins suggest that they may be excellent candidates for elucidating structure-function relationships in biopolymers for nanotechnology and biological applications. PMID- 15877377 TI - Spherulitic crystallization in starch as a model for starch granule initiation. AB - The influence of cooling rate and quench temperature on the formation of spherulitic morphology in heated mung bean starch is reported. Spherulites were obtained for a wide range of cooling rates (2.5-250 degrees C/min), provided the system was heated to 180 degrees C and then cooled below 65 degrees C. Branched crystalline structures were also observed, as was a gellike morphology. The dissolution temperature for spherulitic material ranged between 100 and 130 degrees C. A second dissolution endotherm was observed between 130 and 150 degrees C in systems containing gellike material. Spherulites revealed B-type X ray diffraction patterns. Spherulitic crystallization of starch following phase separation is proposed as a model for starch granule initiation in vivo. PMID- 15877378 TI - Molecular interaction between organophosphorus acid anhydrolase and diisopropylfluorophosphate. AB - Organophosphorus acid anhydrolases (OPAA; E.C.3.1.8.2) are a class of enzymes that hydrolyze a variety of toxic acetylcholinesterase-inhibiting organophosphorus (OP) compounds, including pesticides and fluorine-containing chemical nerve agents. In this paper, subphase conditions have been optimized to obtain stable OPAA Langmuir films, and the diisopropylfluorophosphate (DFP) hydrolysis reaction catalyzed by OPAA in aqueous solution and at the air-water interface was studied. OPAA-DFP interactions were investigated utilizing different spectroscopic techniques, that is, circular dichroism and fluorescence in aqueous solution and infrared reflection absorption spectroscopies at the air water interface. The characterization of OPAA and its secondary structure in aqueous solution and as a monolayer at the air-water interface in the absence and in the presence of DFP dissolved in aqueous solution or in the aqueous subphase demonstrated significantly distinctive features. The research described herein demonstrated that OPAA can be used in an enzyme-based biosensor for DFP detection. PMID- 15877379 TI - Stimuli-responsive surfactant/phospholipid stabilized colloidal dispersions and their film formation. AB - Methyl methacrylate (MMA) and n-butyl acrylate (nBA) were copolymerized into stable colloidal particles in the presence of micelle forming sodium dioctyl sulfosuccinate (SDOSS) and liposome forming 1,2-dilauroyl-sn-glycero-3 phosphocholine (DLPC) in aqueous media that serve as thermodynamically stable loci for lipophilic monomers and nanostructured templates. These studies show for the first time that hollow colloidal particles may coalesce to form polymeric films and the combination of SDOSS and DLPC dispersing agents provides a stimuli responsive environment during film formation through which individual surface stabilizing components can be driven to the film-air (F-A) or film-substrate (F S) interface. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) of p-MMA/nBA colloidal dispersions revealed preferential and enhanced mobility of SDOSS and DLPC lipid rafts to the F-A and F-S interfaces in response to thermal, ionic, and enzymatic stimuli. PMID- 15877380 TI - Effects of protein-polyelectrolyte affinity and polyelectrolyte molecular weight on dynamic properties of bovine serum albumin-poly(diallyldimethylammonium chloride) coacervates. AB - Bovine serum albumin (BSA) and poly(diallyldimethylammonium chloride) (PDADMAC) spontaneously form, over a range of ionic strength I and pH, dense fluids rich in both macroions. To study their nanostructure, these coacervates were prepared at low I and high pH (strong interaction) or at high I and lower pH (weaker interaction), with polymer MWs ranging from 90K to 700K, and then examined by dynamic light scattering (DLS) and rheology. DLS shows a dominant and surprisingly fast protein diffusional mode independent of polymer MW; accompanied by robust slow modes, slower by 1-2 orders of magnitude, which are also insensitive to MW and are present regardless of I, pH, and sample aging. High MW sensitivity was observed by rheology for the terminal time (order of milliseconds), which increased as well with the strength of polyelectrolyte protein interaction. Viscoelastic behavior also indicated a tenuous network, solidlike at low strain but re-forming after breakage by shear. Two models, both of which have strengths and defects, are put forward: (I) macroion-rich domains dispersed in a continuum of macroion-poor domains near the percolation limit and (II) a semidilute solution of PDADMAC chains with interchain friction modulated by transient BSA-PDADMAC association. PMID- 15877381 TI - Morphology of modified regenerated model cellulose II surfaces studied by atomic force microscopy: effect of carboxymethylation and heat treatment. AB - Model cellulose II surfaces with different surface charge have been prepared from carboxymethylated wood pulp. AFM tapping-mode imaging in air showed that the introduction of charged groups into the film does not appreciably change the surface morphology. However, after a mild heat treatment (heating at 105 degrees C for 6 h), an irreversible surface structure change, from near spherical-type aggregates to a fibrillar structure, was observed. This might be attributed to the formation of strong hydrogen bonds in the crystalline region of the films while the amorphous regions shrank upon drying. The suitability of these charged cellulose films for surface forces studies was also investigated. At pH below the pK(a) of the carboxyl groups present in the film, the interaction force could be fit by a van der Waals force interaction. At higher pH, the interaction was of a purely electrostatic nature with no van der Waals component observable due to the swelling of the surfaces. PMID- 15877382 TI - Radicalization of lignocellulosic fibers, related structural and morphological changes. AB - The radicalization of unbleached lignocellulosic fibers obtained from thermomechanical (TMP) and chemothermomechanical (CTMP) pulps was performed in heterogeneous phase by reaction with dioxygen in the presence of N,N' ethylenebis(salicylideneiminato)cobalt(II), [Co(salen)], as catalyst. Phenoxy cobalt radicals immobilized in fibers were observed by electron paramagnetic resonance (EPR) spectroscopy; their amount depends on the fiber swelling induced by reaction medium. The absolute concentration of such radicals in fibers, about 10(16) spin/g, reaches values 10 times higher than that of phenoxy radicals formed in similar oxidative reactions catalyzed by laccase. The generation of phenoxy cobalt radicals in fibers was related to structural changes of lignin units, detected by mono- and bidimensional nuclear magnetic resonance ((13)C NMR and 2D-HSQC) investigations, and to morphological modifications in fibers observed by scanning electron microscopy (SEM). PMID- 15877383 TI - Protein-functionalized polymer brushes. AB - A new strategy for the preparation of protein-functionalized polymer brushes is reported, which is based on a combination of surface-initiated atom transfer radical polymerization (ATRP), p-nitrophenyl chloroformate activation of the surface hydroxyl groups, and subsequent O(6)-benzylguanine (BG) functionalization. The BG-functionalized brushes are used to chemoselectively immobilize O(6)-alkylguanine-DNA-alkyltransferase (AGT) fusion proteins with a defined orientation and surface density. These protein-modified polymer brushes are attractive candidates for the development of protein microarrays. PMID- 15877384 TI - Encapsulation of osteoblast seeded microcarriers into injectable, photopolymerizable three-dimensional scaffolds based on d,l-lactide and epsilon caprolactone. AB - UMR-106 seeded microcarriers were encapsulated into in situ, photopolymerizable three-dimensional scaffolds based on d,l-lactide and epsilon-caprolactone. UMR 106 and rat bone marrow cells proliferated and differentiated well on the microcarriers. The microcarriers were completely colonized after 14 days in culture. The viscous polymer paste allowed to mix the UMR-106 seeded microcarriers and gelatin (porosigen) properly. After the photopolymerization process, microcarriers and gelatin were evenly distributed throughout the scaffold. Gelatin was leached out within 7 h, and a porous scaffold was obtained. The microcarriers remained in the scaffold even after 7 days which demonstrates that they were well entrapped in the polymer. Increasing the amount of entrapped microcarriers (20-50%) leads to scaffolds with a reduced cross-linking. Hence, the microcarriers leached out. The encapsulated UMR-106 cells did not show pyknotic nuclei which demonstrates that the photopolymerization and handling the viscous polymer/gelatin/microcarrier paste is not detrimental for the cells. PMID- 15877385 TI - In situ formation of blends by photopolymerization of poly(ethylene glycol) dimethacrylate and polylactide. AB - Blends of cross-linked poly(ethylene glycol) dimethacrylate (PEGDMA) and poly(d,l lactide) (PLA) were prepared by mixing photoactive PEGDMA (molecular mass: 875 g/mol) and PLA, and subsequently photopolymerizing the mixture with visible light. The effects of PLA molecular mass and mass fraction on the rheological properties of the PEGDMA/PLA mixtures, and on the degree of methacrylate vinyl conversion (DC), as well as blend miscibility, microstructure, mechanical properties, in vitro swelling behavior, and cell responses were studied. PLA-2K (molecular mass: 2096 g/mol) and PLA-63K (molecular mass: 63 000 g/mol) formed miscible and partially miscible blends with cross-linked PEGDMA, respectively. The addition of the PLA-2K did not affect the immediate or post-cure (>24 h) DC of the PEGDMA upon photopolymerization. However, the addition of PLA-63K decreased the immediate DC of the PEGDMA, which can be increased through extending the curing time or post-curing period. Compared to the cross-linked neat PEGDMA and PLA-2K/PEGDMA blends, PLA-63K/PEGDMA blends were significantly stronger, stiffer, and tougher. Both types of blends and the cross-linked PEGDMA swelled when soaked in a phosphate buffered saline (PBS) solution. The attachment and spreading of MCT3-E1 cells increased with increasing PLA-63K content in the blends. The facile and rapid formation of PEGDMA/PLA blends by photopolymerization represents a simple and efficient approach to a class of biomaterials with a broad spectrum of properties. PMID- 15877386 TI - Flory temperature and upper critical solution temperature of gelatin solutions. AB - The Flory temperatures (theta) measured by turbidity experiments performed on gelatin solutions were found to be 12 +/- 0.3, 13 +/- 0.3, 14 +/- 0.3, 14.5 +/- 0.3, and 15 +/- 0.3 degrees C for salt concentrations 0.1, 0.075, 0.05, 0.025, and 0 M (NaCl), respectively. Estimated persistence length (l(p)) of this weakly charged polyelectrolyte could be deduced from the Benoit and Doty (J. Phys. Chem. 1953, 57, 958) relationship with the approximation that this biopolymer assumes a compact near-globular shape at Flory temperature, implying l(p) = 9(R(h))(2)/(5L(m)), where L(m) is the contour length and R(h) is the hydrodynamic radius. It was found that l(p) approximately 2.2 +/- 0.2 nm at room temperature (20 degrees C), invariant of salt concentration. The Flory expansion factor (alpha= R(h)(T)/R(h)(theta) = 1.5+/-0.2) was found to be almost constant. theta Composition for this biopolymer was deduced from turbidimitric titration of aqueous gelatin solutions with the alcohols methanol, ethanol, 2-propanol, and tert-butyl alcohol. It appears that hydrophobic interactions play a crucial role in causing chain collapse at theta-temperature and composition. PMID- 15877387 TI - Covalent immobilization of cellulose layers onto maleic anhydride copolymer thin films. AB - Thin films of cellulose are advantageous for analytical studies in aqueous environments to investigate various factors determining the performance of cellulose-based products. However, the weak fixation of cellulose layers on common carrier materials often limits this approach. To address this problem, we suggest a novel maleic anhydride copolymer precoating technique which allows for the covalent attachment of cellulose thin films through esterification. Maleic anhydride copolymers were deposited and covalently bound onto planar, aminosilane modified glass or silicon oxide surfaces. Cellulose was subsequently immobilized on top of the copolymer precoatings by spin coating from N-methylmorpholine-N oxide/dimethyl sulfoxide solutions. The resulting cellulose films were thoroughly characterized with respect to layer thickness, morphology, chemical constitution, and electrical charging. The stability of the layers against shear stress was demonstrated in aqueous solutions and the covalent attachment of the cellulose to the copolymer films was proven by means of dissolution experiments followed by ellipsometry and high-resolution X-ray photoelectron spectroscopy. PMID- 15877388 TI - Structure-property relationships in poly(ethylene glycol)-protein hydrogel systems made from various proteins. AB - A series of poly(ethylene glycol)-protein hydrogels were synthesized with different proteins, and the resultant structures were characterized in terms of swelling behavior and mechanical, optical, and drug release properties. Irrespectively of the protein involved in polymerization with poly(ethylene glycol), all studied systems were found to be loosely cross-linked networks, where both polymer and protein are completely solvated, enabling as high as 96% water content. Changes in the apparent transparency of the hydrogels synthesized with different proteins were attributed to the ability of the protein component to self-associate via hydrophobic interactions. The polyelectrolyte nature of the protein component governs the pH responsiveness of the network, which manifested itself in a pH-dependent mechanism of swelling and drug release. It was demonstrated that there is great opportunity to modulate the final characteristics of the hydrogel system to fit the need of specific biomedical application. PMID- 15877389 TI - Formation of polyelectrolyte complex particles from self-complexation of N sulfated chitosan. AB - This work investigated the elaboration of biocompatible nanoparticles from the pH induced self-complexation of the amphoteric polysaccharide N-sulfated chitosan. The acidification of aqueous solutions of chitosan having a degree of acetylation of 24% and a degree of sulfation of 34% or 56% was followed stepwise by turbidimetry, dynamic light scattering, and electrophoresis. With the highest sulfated chitosan, no turbidity was recorded between pH = 7.8 and 2.0, traducing a high apparent solubility of the polymer chains in this domain of pH. With the lowest sulfated chitosan, a steady increase in turbidity was monitored from pH = 6.90 to 6.15 followed by the flocculation of the polymer at pH approximately 6.0. In this range of pH, the polymer phase separated to yield particles having hydrodynamic diameters decreasing from 350 to 260 nm and an almost constant negative charge. These particles were assembled by electrostatic interactions between the protonated amino residues and the sulfate functions and stabilized by an excess of surface sulfate groups. The particles could be separated from the reaction medium and concentrated by centrifugation-redispersion cycles without alteration of their structure. PMID- 15877390 TI - Characterization of methylprednisolone esters of hyaluronan in aqueous solution: conformation and aggregation behavior. AB - Methylprednisolone steroid esters of hyaluronan differing in degree of functionalization and molecular weight were investigated in aqueous solution. Conformation and aggregation phenomena were elucidated by means of circular dichroism, viscometry, rheology, and nuclear magnetic resonance, mainly by (1)H pulsed field gradient (PFG) NMR, which allows the determination of the diffusion coefficient of the species under investigation. The functionalization of hyaluronan with the steroid induces a reduction of the molecular volume, as a consequence of intramolecular hydrophobic interactions. For concentrated samples we have observed the coexistence of unimolecular collapsed chains and of aggregates, the latter disappearing upon dilution. The methylprednisolone ester of lower molecular weight hyaluronan has a larger molecular volume than its higher molecular weight analogue, even though still smaller than the underivatized polymer. This effect can be explained with the reduced flexibility of the polymer backbone probably impairing intramolecular interactions. PMID- 15877391 TI - Role of the pericarp cellulose matrix as a moisture barrier in microwaveable popcorn. AB - Since moisture loss of popcorn can cause an increase in the number of unpopped kernels, pericarp properties of popcorn hybrids were analyzed to understand factors affecting moisture loss rate during microwave heating. Differential scanning calorimetry profiles of ground pericarp displayed a notable exothermal event, and hybrids with superior microwave popping performance (fewer unpopped kernels) exhibited significantly higher enthalpies. The number of unpopped kernels was highly correlated (r = 0.826, p = 0.011) with pericarp enthalpy values. X-ray analysis confirmed that cellulose and arabinoxylan are the major structural components of the pericarp. Structural changes in cellulose were induced by moisture and heat, and considerable enhancement in crystallinity occurred when the pericarp was heated in the presence of water. Results of this study indicate that the cellulose component of the pericarp is responsible for the development of exothermal events and increased crystallinity. Thus, the propensity of cellulose to form crystalline structures in the popcorn pericarp during microwave heating improves moisture retention and hence popping performance. PMID- 15877392 TI - Intermolecular interactions and phase structures of plasticized wheat proteins materials. AB - The intermolecular interactions and phase structures of thermally processed wheat proteins with glycerol and water as plasticizers were studied by dynamic mechanical analysis and solid-state high-resolution NMR spectroscopy. The results of phase structures at scales of molecular level to tens of nanometers were correlated with the mechanical properties of the materials. The strong hydrogen bonding intermolecular interactions between the components in wheat proteins and the plasticizers resulted in a significant change in molecular motions of wheat protein materials. The plasticized systems, however, still presented a wide distribution of chain mobility at a scale from the molecular level to 20-30 nm, and the plasticizing effect was different for each wheat protein system. High protein content systems tended to be plasticized relatively easily especially when lipid content is high, but the existence of residual starch would require more plasticizers to reach a similar level of chain mobility. On a scale of 20-30 nm, plasticized vital wheat gluten (WG) and the deamidated wheat proteins (WP-I) were heterogeneous with each component exhibiting its individual mobility, whereas the plasticized insoluble protein system (WP-II) with poor mechanical properties was homogeneous. Both WG and WP-I systems showed excellent mechanical polymeric properties in tensile strength and elasticity despite the heterogeneity. The strong intermolecular hydrogen bonding interactions and soluble protein components in the materials could provide an adhesion among different components and act as a continuous matrix in the systems. Therefore, these materials displayed excellent mechanical properties via coordination effects among different components. PMID- 15877393 TI - Purification and characterization of heparin from the Italian clam Callista chione. AB - An unusual heparin (approximately 1.9 mg/g of dry tissue) was isolated from the marine italian bivalve mollusk Callista chione. Agarose gel electrophoresis showed a high content of the fast-moving heparin component (85 +/- 7.6%) and 15 +/- 1.3% of the slow-moving species. An average molecular mass of 10 950 was calculated by PAGE analysis. The anticoagulant properties were measured as APTT (97 +/- 12.1 IU/mg) and anti-Xa activity (52 +/- 7.4 IU/mg). Structural analysis of clam heparin, performed by depolymerizing heparin samples with heparinase (EC 4.2.2.7) and then separating the resulting unsaturated oligosaccharides by SAX HPLC, revealed the presence of low amounts of the trisulfated disaccharide [DeltaUA2S(1-->4)-alpha-d-GlcN2S6S] and a significant increase of the disaccharides bearing nonsulfated iduronic and glucuronic acids, [-->4)-alpha-l IdoA(1-->4)-alpha-d-GlcNAc6S(1-->] and [-->4)-alpha-l-IdoA(1-->4)-alpha-d GlcN2S6S(1-->], and [-->4)-beta-d-GlcA(1-->4)-alpha-d-GlcN2S6S(1-->]. As a consequence, Callista chione heparin is a low-sulfated polysaccharide showing a specific decrease of the sulfatation in position 2 of the uronic acid units. PMID- 15877394 TI - Macrolactones and polyesters from ricinoleic acid. AB - A systematic study on the synthesis, characterization, and polymerization of ricinoleic acid (RA) lactone is reported. Ricinoleic acid lactones were synthesized by refluxing pure ricinoleic acid in chloroform (10 mg/mL) with dicyclohexylcarbodimide and (dimethylamino)pyridine as catalyst. Purification of RA lactones was performed by silica gel chromatography. The reaction resulted in a 75% yield of ricinoleic acid lactones. IR and NMR analysis confirmed the formation of cyclic compounds. Polymerization of the ricinoleic acid lactones with catalysts commonly used for ring-opening polymerization of lactones, under specific reaction conditions, resulted in oligomers. Copolymerization with lactide (LA) by ring-opening polymerization, using Sn(Oct) as catalyst, yielded copolyesters with molecular weights (M(w)) in the range of 5000-16000 and melting temperatures of 100-130 degrees C for copolymers containing 10-50% w/w ricinoleic acid residues. Degradation studies of the copolymers were performed in 0.1 M phosphate buffer solution, pH 7.4, at 37 degrees C. P(LA-RA)s with up to 20% w/w RA slowly degraded and released only approximately 7% of its lactic acid content after 60 days of study, while pure PLA under similar conditions released more than 20% of its lactic acid content. On the other hand, copolyesters containing more then 20% w/w RA degraded and released lactic acid faster than pure PLA due to the low crystallinity of the copolymers. PMID- 15877395 TI - Patterning poly(organophosphazenes) for selective cell adhesion applications. AB - Five polyphosphazenes with different hydrophilicites were synthesized and screened in vitro. The purpose was to identify unique types of polymeric substrates that distinctly favored or markedly prevented cellular adhesion. The SK-N-BE(2c) human neuroblastoma cell line, utilized for its electrogenic responses, was used to test this differential adhesion. In particular, the objective was to specifically culture this cell line in a highly selective pattern. Each candidate polymer was cast into films and plated with neuroblastoma cells for 3 days. The polyphosphazene materials which showed negative cellular adhesive properties (-CAPs) were poly[bis(trifluoroethoxy)phosphazene] (TFE) and poly[bis(methoxyethoxyethoxy)phosphazene] (MEEP). The polyphosphazenes which showed positive cellular adhesive properties (+CAPs) were poly[(methoxyethoxyethoxy)(1.0)(carboxylatophenoxy)(1.0)phosphazene] (PMCPP), poly[(methoxyethoxyethoxy)(1.0)(cinnamyloxy)(1.0)phosphazene] (PMCP), and poly[(methoxyethoxyethoxy)(1.0)(p-methylphenoxy)(1.0)phosphazene] (PMMP). To test cellular selectivity, films of -CAP and +CAP were copatterned onto glass substrates. The micropatterned films were plated with SK-N-BE(2c) neuroblastoma cells for one week. The results showed that neuroblastoma cells adhere selectively (over 60%) to the +CAP microfeatures. We also showed that multiple properties can be achieved with a single material and that we can use TFE as both a -CAP and an insulation layer and PMCP as a conductive +CAP layer. PMID- 15877396 TI - Electropolymerized films formed from the amphiphilic decyl esters of D- and L tyrosine compared to L-tyrosine using the electrochemical quartz crystal microbalance. AB - Using the electrochemical quartz crystal microbalance (EQCM), we compared thin films formed on Pt by electropolymerization of l-tyrosine to that of the amphiphilic monomers, decyl esters of d- and l-tyrosine (DEDT and DELT). Mass build-up and film properties were determined as a function of monomer concentration via frequency, f, motional resistance, R, and charge passage, Q, measurements. Films were found to occur by a combination of monomer electropolymerization and adsorption for DEDT and DELT, but only by electropolymerization for l-tyrosine. This difference in film formation process for the monomers is reflected in the net mass build-up for each film, as represented by calculated df/dQ values. For the adsorbing monomers DEDT and DELT, films possessed concentration dependent df/dQ values, more than 100-fold greater than that for l-tyrosine film formation under equivalent electropolymerization conditions. During the entire film growth process, all three films exhibited no significant energy dissipation properties (DeltaR invariant). Concentration dependent adsorption of significant levels of unpolymerized but self-assembled DEDT and DELT monomers account for the subsequent time dependent mass loss observed from the films maintained in buffer in the absence of monomer. Contact angle measurements demonstrated a pH dependent increase in the surface hydrophilicity of films electropolymerized from the DEDT, DELT, and l-tyrosine monomers but not films formed from phenol and 3-nitrophenol monomers. This behavior is consistent with the monomers' known changes in titration/charge state properties with increasing pH. This study provided insight into the film formation, stability, and surface hydrophilicity resulting from electropolymerization of these related tyrosine based monomers. This information is critical to assessing the utility these films may have in the development of new biomaterials and as biological macromolecule or cell immobilization strategies in biosensors. PMID- 15877397 TI - Asymmetric epoxidation of alpha-olefins having neighboring sugar chiral templates and alternating copolymerization with dicarboxylic anhydrides. AB - Sugar-substituted epoxides 5-8 were synthesized by asymmetric epoxidation (in CH(2)Cl(2)/water) of alpha-olefins having neighboring sugars (1-4) by use of an achiral oxidant (MCPBA), in which the sugar moiety acted as a chiral template. The diastereoselectivities depend on the methylene spacer between vinyl group and carbohydrate derivatives. The methylene spacer between sugar and vinyl groups influenced the diastereoselectvity. In the case of epoxidation of 4 at 27 degrees C for 24 h, the diastereoselectivity was the highest (99/1). Copolymerizations of 5-8 with succinic anhydride were attained at 100 degrees C for 72 h to give poly(ethylene succinate) having pendant carbohydrate [poly(SAn-alt-5), M(n) = 1.4 x 10(3); poly(SAn-alt-6), M(n) = 2.2 x10(3); poly(SAn-alt-7), M(n) = 2.9 x 10(3); poly(SAn-alt-8), M(n) = 1.8 x 10(3)]. The methylene spacer between sugar and epoxide has an effect on the reactivity of epoxide in copolymerization as well as the diastereoselectivity. Alternating copolymerization of 7 and glutaric anhydride gave a polyester of M(n) 4.2 x10(3). PMID- 15877398 TI - Synthesis and characterization of novel blood-compatible soluble chemically cross linked polyurethanes with excellent mechanical performance for biomedical applications. AB - A controlled cross-linking polymerization system was designed, and soluble chemically cross-linked polyurethane was synthesized using laurylamine, n octylamine, n-pentylamine, and ethylenediamine chain extenders. The mechanical analysis showed that the polyurethane materials synthesized in this paper have very excellent mechanical properties with a breaking elongation of 1914% and a tensile strength of 4303 N/cm(2). Such good mechanical properties must enable it to have good longevity when used as biomaterials. The polyurethane materials with n-pentylamine and n-octylamine chain extenders show reduced platelet adhesion than that with an ethylenediamine chain extender after sustaining 200 000 times of load cycles, indicating that polyurethanes introduced with an alkyl side chain onto the hard segments keep good antithrombogenic properties after sustaining load cycles. This might be because the hard segments are shielded by the alkyl side chain when the micro-phase-separation structure is destroyed in the repeated deformation of the polyurethane materials. The present investigation reveals that the influence of introducing long alkyl side chains into the backbone of the polyurethane macromolecule has been shown to reduce platelet deposition and to enhance in vitro albumin adsorption. However, in this paper, it has been observed that the polyurethane material introduced with a proper-length alkyl side chain onto the hard segment has the best antithrombogenic properties after the fatigue test. PMID- 15877399 TI - Structural studies of Bombyx mori silk fibroin during regeneration from solutions and wet fiber spinning. AB - Regenerated silk fibroin materials show properties dependent on the methods used to process them. The molecular structures of B. mori silk fibroin both in solution and in solid states were studied and compared using X-ray diffraction, FTIR, and (13)C NMR spectroscopy. Some portion of fibroin protein molecules dissolved in formic acid already have a beta-sheet structure, whereas those dissolved in TFA have some helical conformation. Moreover, fibroin molecules were spontaneously assembled into an ordered structure as the acidic solvents were removed from the fibroin-acidic solvent systems. This may be responsible for the improved physical properties of regenerated fibroin materials from acidic solvents. Regenerated fibroin materials have shown poor mechanical properties and brittleness compared to their original form. These problems were technically solved by improving the fiber forming process according to a method reported here. The regenerated fibroin fibers showed much better mechanical properties compared to the native silk fiber and their physical and chemical properties were characterized by X-ray diffraction, solid state (13)C NMR spectroscopy, SinTech tensile testing, and SEM. PMID- 15877400 TI - Analysis using a linear viscoelastic model of the in vitro osmotic kinetics of polydisperse synthetic colloids. AB - This study clarifies the contribution to overall osmotic kinetics of colloid osmotic pressure (Pi) and the interaction of synthetic colloids with the membrane. Solutions (6%) of dextran with weight average molecular weight (MW(w)) 68 800 (DEX 70), dextran with MW(w) 40 000 (DEX 40), hydroxyethyl starch with MW(w) 70 000 (HES 70), gelatin with MW(w) 60 000 and albumin were tested. An osmotic flow cell fitted with membranes of molecular weight cutoff size 30 000 or 50 000 was used to measure time-dependent changes in Pi for each of these solutions. A linear viscoelastic model was fitted to the curve describing changes to Pi as a function of time. Values of total effective Pi for DEX 40 and DEX 70 were larger than those for HES 70, gelatin, and albumin. As an index of solute solvent exchange rate at the membrane surface, these values were in the order DEX 40 > DEX 70, HES 70 > gelatin, albumin. The findings suggest that DEX 40 may be preferable for the temporary restoration of plasma volume because of a heightened initial osmotic force. In contrast, the osmotic force exerted by gelatin is slower to increase but is likely to be longer lasting in vivo as a result of the inhibition of gelatin from penetrating the capillary membrane due to its interaction with negatively charged groups in the endothelial glycocalyx. PMID- 15877401 TI - Reversible hydrogels from self-assembling genetically engineered protein block copolymers. AB - A series of triblock protein copolymers composed of a central water-soluble polyelectrolyte segment flanked by two coiled-coil domains was synthesized by genetic engineering methods. The copolymers self-assembled into reversible hydrogels in response to changes in temperature, pH, and the presence or absence of denaturating agent (guanidine hydrochloride, GdnHCl). Hydrogel formation was concentration-dependent, and the concentration needed for hydrogel formation correlated with the oligomerization state of the coiled-coil domains in the protein copolymers. The morphology of the hydrogels, as determined by scanning electron microscopy (SEM), indicated the presence of porous interconnected networks. The thermal stabilities and self-assembling properties of the protein copolymers were successfully controlled by manipulating the amino acid sequences of the coiled-coil domains. The stimuli responsiveness and reversibility of the hydrogel self-assembly suggest that these protein copolymers may have potential in biomedical applications. PMID- 15877402 TI - Synthesis and characterization of polymeric soybean oil-g-methyl methacrylate (and n-butyl methacrylate) graft copolymers: biocompatibility and bacterial adhesion. AB - Peroxidation, epoxidation, and/or perepoxidation reactions of soybean oil under air at room temperature resulted in cross-linked polymeric soybean oil peroxides on the surface along with the waxy soluble part, sPSB, with a molecular weight of 4690, containing up to 2.3 wt % peroxide. This soluble polymeric oil peroxide, sPSB, initiated the free radical polymerization of either methyl methacrylate (MMA) or n-butyl methacrylate (nBMA) to give PSB-g-PMMA and PSB-g-PnBMA graft copolymers. The polymers obtained were characterized by (1)H NMR, thermogravimetric analysis, differential scanning calorimetry, and gel permeation chromatography techniques. Polymeric oil as a plasticizer lowered the glass transition of the PSB-g-PMMA graft copolymers. PSB-g-PMMA and PSB-g-PnBMA graft copolymer film samples were also used in cell culture studies. Fibroblast and macrophage cells were strongly adhered and spread on the copolymer film surfaces, which is important in tissue engineering. Bacterial adhesion on PSB-g-PMMA graft copolymer was also studied. Both Staphylococcus epidermidis and Escherichia coli adhered on the graft copolymer better than on homo-PMMA. Furthermore, the latter adhered much better than the former. PMID- 15877403 TI - Heparin-coupled poly(poly(ethylene glycol) monomethacrylate)-Si(111) hybrids and their blood compatible surfaces. AB - Well-defined (nearly monodispersed) poly(poly(ethylene glycol)monomethacrylate) Si hybrids were prepared via surface-initiated atom transfer radical polymerization (ATRP) of the poly(ethylene glycol)monomethacrylate (PEGMA) macromonomer on the hydrogen-terminated Si(111) surface (Si-H surface). Both the active chloride groups at the chain ends (from the ATRP process) and the chloride groups converted from some ( approximately 32%) of the -OH groups of the Si-C bonded PEGMA polymer, or P(PEGMA), brushes were used as leaving groups for the covalent coupling of heparin. For the heparinized P(PEGMA)-Si hybrid surfaces, protein adsorption and platelet adhesion were significantly suppressed. The well defined and dense P(PEGMA) brushes, prepared from surface-initiated ATRP, had allowed the immobilization of a relatively high concentration of heparin (about 14 mug/cm(2)). The resulting silicon surface exhibited significantly improved antithrombogenecity with a plasma recalcification time (PRT) of about 150 min. The persistence of high bioactivity for the immobilized heparin on the hybrid surfaces can be attributed to the biocompatibility of the PEGMA units, as well as their role as spacers in providing the immobilized heparin with a higher degree of conformational freedom in a more hydrophilic environment. Thus, the heparin coupled P(PEGMA)-Si hybrids with anti-fouling and antithrombogenic surfaces are potentially useful in silicon-based implantable devices and tissue engineering. PMID- 15877404 TI - Synthesis and characterization of new permanently charged poly(amidoammonium) salts and evaluation of their DNA complexes for gene transport. AB - A new series of linear and permanently charged poly(amidoammonium) salts were synthesized in order to investigate the influence of their ionic and hydrophobic contents on both the cytotoxicity and the transfection mediated by polycation-DNA complexes. The poly(amidoammonium) salts were prepared by chemical modification of a parent poly(amidoamine) containing two tertiary amino groups per structural unit: one incorporated into the main chain and the other fixed at the end of a short bismethylene spacer. The permanent charges were introduced through a quaternization reaction involving iodomethane or 1-iodododecane as an alkylating agent. Under appropriate conditions, the methylation reaction was found to be regioselective, allowing the quaternization of either the side chains or both the side chains and the backbone. Under physiological salt conditions (150 mM NaCl), all of the poly(amidoammonium) salts self-assembled with DNA to form complexes. High proportions of highly quaternized polycation provided better defined morphology to the polycation-DNA complexes. Complexes formed from unquaternized polycation were less cytotoxic than branched poly(ethyleneimine) (25 kDa). At high polycation-DNA weight ratios, the introduction of permanent charges generated a significant increase in the cytotoxicity, but no patent correlation could be established with the amount and the position of the permanent charges. Only complexes formed from polycations with quaternized backbone were able to generate significant gene expression, which was putatively attributed to a better defined toroidal-like morphology together with a higher stability, as suggested by zeta potential measurements. The incorporation of dodecane side chains on highly charged polycations severely amplified the cytotoxicity so that, in return, the transfection level was dramatically affected. PMID- 15877405 TI - Multilayers of a globular protein and a weak polyacid: role of polyacid ionization in growth and decomposition in salt solutions. AB - Thin films obtained from a layer-by-layer deposition of a weak polycarboxylic acid and a positively charged globular protein were studied by in situ ATR-FTIR. The system was chicken egg lysozyme (Lys), bovine pancrease ribonuclease A (RNase), or bovine gamma-globulin (IgG) self-assembled with polycarboxylic acids. When the pH value was lowered below a critical point, the growth of films and their tolerance to decomposition by added sodium chloride improved dramatically. Stabilization of protein/polyacid films in salt solutions at lower pH values occurred due to the onset of nonelectrostatic interactions to intermolecular binding within protein/polyacid multilayers and was controlled by polyacid ionization within the film rather than the pH of the external solution. A fractional ionization of polyacid in the pH-stabilization region was lower with protein-containing films than for polyacid/linear polycation films, reflecting hindrance of the inter-association of protonated carboxylic groups by protein globules. Practical ramifications of the pH-stabilization effect might extend to areas of biotechnology and biomaterials. PMID- 15877406 TI - Preparation and properties of plasticized poly(lactic acid) films. AB - Poly(lactic acid), PLA, was blended with monomeric and oligomeric plasticizers in order to enhance its flexibility and thereby overcome its inherent problem of brittleness. Differential scanning calorimetry, dynamic mechanical analysis, transmission electron microscopy, and tensile testing were used to investigate the properties of the blends. Monomeric plasticizers, such as tributyl citrate, TbC, and diethyl bishydroxymethyl malonate, DBM, drastically decreased the T(g) of PLA, but the blends showed no morphological stability over time since rapid cold crystallization caused a size reduction of the amorphous domains in PLA. Consequently, the ability of PLA to accommodate the plasticizer diminished with the increase in crystallinity and migration of the plasticizer occurred. Increasing the molecular weight of the plasticizers by synthesizing oligoesters and oligoesteramides resulted in blends that displayed T(g) depressions slightly smaller than with the monomeric plasticizers. The compatibility with PLA was dependent on the molecular weight of the oligomers and on the presence or not of polar amide groups that were able to positively interact with the PLA chains. Aging the materials at ambient temperature revealed that the enhanced flexibility as well as the morphological stability of the films plasticized with the oligomers could be maintained as a result of the higher molecular weight and the polar interactions with PLA. PMID- 15877407 TI - Cell engineering biointerface focusing on cytocompatibility using phospholipid polymer with an isomeric oligo(lactic acid) segment. AB - Initial contact between a biological environment and a biomaterial ultimately decides the in vivo performance. Therefore, the fabrication of a delicate biointerface is important because it can be utilized as a platform for novel biomaterials. For the preparation of advanced biomedical devices such as biochips, nanoparticles, and cell engineering devices, the surface properties may be modified by the design of polymeric biomaterials. Anomalous phospholipid polymers with an isomeric oligo(lactic acid) segment were designed and evaluated as a biointerface. The phospholipid polymer containing 2-methacryloyloxyethyl phosphorylcholine was easily copolymerized with isomeric oligo(lactic acid) macromonomers, and the obtained polymer could easily form thin coating membranes as biointerfaces. The oligo(lactic acid) involves three kinds of isomers: dl-, d , and l-forms. The favorable characteristic on the surface provides regulation of cell-material interactions on the biointerface. The oligo(lactic acid) segment could form hydrophobic domains, which were considered to be located on the interface, to enhance protein adsorption and cell adhesion. The most favorable characteristics on the biointerface were dual functions of cytocompatibility by the phospholipid polymer and cell adhesion property by the oligo(lactic acid) segment. In this study, we focused on the biological responses such as protein adsorption and cell adhesion by change in the oligo(lactic acid) component. The cell viability on the confluent stage was evaluated in terms of metabolic activity. PMID- 15877408 TI - Time-resolved X-ray diffraction study on poly[(R)-3-hydroxybutyrate] films during two-step-drawing: generation mechanism of planar zigzag structure. AB - Uniaxially oriented films with high tensile strength were processed from ultrahigh-molecular-weight poly[(R)-3-hydroxybutyrate] (P(3HB)) by a method combining hot-drawing near the melting point of the polymer and two-step-drawing at room temperature. In a two-step-drawn and subsequently annealed film, P(3HB) molecular chains fall into two states: 2/1 helix (alpha-form) and planar zigzag (beta-form) conformations. The mechanism for generating the beta-form during two step-drawing was investigated by time-resolved synchrotron wide- and small-angle X-ray scattering measurements (WAXD and SAXS), together with the measurement of stress-strain curves. It was found that the improvement of mechanical properties is due to not only the orientation of molecular chains but also the generation of the beta-form during the drawing. The crystal and molecular structures of the alpha-form remained unchanged until the yield point of the stress-strain curve. At the yield point, the long period obtained from SAXS doubled and a new reflection indicative of the beta-form was observed on the equatorial line in WAXD. The intensity of the reflection from the beta-form increased with an increase in the two-step-drawing ratio at room temperature. The SAXS pattern changed from a two-point reflection along the meridian to a cross pattern with streaking on the equatorial line, demonstrating the close alignment of shish kebab structures. The reflection intensity, crystal orientation and crystal size of the alpha-form decreased during two-step-drawing. Based on these results, the beta-form is mainly introduced from the orientation of free molecular chains in the amorphous regions between alpha-form lamellar crystals, but the structural transformation of molecular chains also occurs from the alpha-form to the beta form at the deformed lamellar crystals. PMID- 15877409 TI - Synthesis of water-soluble electroactive ferrocene-grafted poly(p-phenylene ethynylene) via phase transfer and its biosensing application. PMID- 15877410 TI - Stretching single molecules of connective tissue glycans to characterize their shape-maintaining elasticity. PMID- 15877411 TI - A monomeric thallium(I) amide in the solid state: synthesis and structure of TlN(Me)ArMes2 (ArMes2 = C6H3-2,6-Mes2). AB - Reaction of TlCl and [LiN(Me)Ar(Mes)2](2) [Ar(Mes)2 = C(6)H(3)-2,6-(C(6)H(2) 2,4,6-Me(3))(2)] in Et(2)O generated the thallium amide, TlN(Me)Ar(Mes)2 (1). X ray data showed that it has a monomeric structure with an average Tl-N distance of 2.364(3) Angstroms. There was also a Tl-arene approach [Tl-centroid = 3.026(2) Angstroms (avg)] to a flanking mesityl ring from the terphenyl substituent. DFT calculations showed that this interaction is weak and supported essentially one coordination for thallium. The electronic spectrum of 1 is hypsochromically shifted in comparison to the monomeric TlAr(Trip)2 (Trip = C(6)H(2)-2,4,6 Pr(i)(3)). PMID- 15877412 TI - A unique cyanide-bridged three-dimensional (3-D) copper(II)-copper(I) mixed valence polymer containing 1-D water tapes with cyclic pentamer units. AB - A unique cyano-bridged copper(II)-copper(I) mixed-valence polymer, {[Cu(3)(CN)(4)(H(2)O)(3)](H(2)O)(2)}(n) (1), has been prepared and structurally determined by X-ray diffraction. The cyanide anions with (mu(2),eta(2)) and (mu(3),eta(2)) bridging modes connect the copper centers into a complicated zeolite-like 3-D network. Of further interest, coordinated and lattice water molecules in this structure form 1-D hydrogen-bonded water tapes consisting of linked cyclic pentamer clusters. PMID- 15877413 TI - The first cobalt metallacrowns: preparation and characterization of mixed-valence cobalt(II/III), inverse 12-metallacrown-4 complexes. AB - Aerobic reactions of Co(O(2)CMe)(2).4H(2)O with di-2-pyridyl ketone oxime (Hpko) in the presence of counterions (ClO(4)(-), PF(6-)) give the tetranuclear, mixed valence cobalt(II/III) clusters [Co(II)(2)Co(III)(2)(OR)(2)(O(2)CMe)(2)(pko)(4)S(2)]X(2) [R = H, S = MeOH, X = ClO(4) (1); R = Me, S = EtOH, X = PF(6) (2)] depending on the solvent mixture. Complexes 1 and 2 are the first Co members in the family of metallacrowns adopting the extremely rare inverse 12-metallacrown-4 motif. PMID- 15877414 TI - One-dimensional chain of tetranuclear manganese single-molecule magnets. AB - A one-dimensional chain of interconnected single-molecule magnets (SMMs) is obtained that consists of [Mn(4)(hmp)(6)](4+) units bridged by chloride ions. Slow magnetization relaxation is evident in the AC susceptibility data and in magnetization hysteresis measurements for [Mn(4)(hmp)(6)Cl(2)](n)(ClO(4))(2)(n). The magnetization hysteresis loops for this complex are similar to those for an SMM and show significant coercive field and steps at regular magnetic intervals. Spin-canted antiferromagnetic coupling due to misalignment of easy axes of neighboring Mn(4) units is also observed for this complex. PMID- 15877415 TI - A straightforward synthesis of diverse nickel dithiolene complexes appended with hydrogen-bond donor/acceptor groups. AB - Nickel dithiolene complexes symmetrically appended with hydrogen-bond donor/acceptor functionalities such as imide, amide, or cyano/amide groups have been synthesized by a straightforward method of wide applicability starting from one single precursor. Single-crystal X-ray structures reveal the occurrence of one type of ribbon common to all compounds that is based on a recurrent self complementary intermolecular hydrogen-bonded ring motif linking the symmetrically substituted complexes. PMID- 15877416 TI - Evidence for static quenching of MLCT excited states by iodide. AB - The metal-to-ligand charge-transfer (MLCT) excited states of Ru(deeb)(bpy)(2)(PF(6))(2) [where bpy is 2,2-bipyridine and deeb is 4,4' (CO(2)CH(2)CH(3))(2)-2,2'-bipyridine] in acetonitrile or dichloromethane were found to be quenched by iodide at room temperature. The ionic strength dependence of the optical spectra gave evidence for ion pairing. Iodide is found to quench the photoluminescence (PL) intensity and influence the spectral distribution of the emitted light. A static component to the time-resolved PL quenching provided further evidence for ground-state adduct. Stern-Volmer analysis of the static component provided an estimate of the iodide-Ru(deeb)(bpy)(2)(2+) adduct equilibrium constant in dichloromethane, K(sv) = 40,000 M(-)(1). Transient absorption studies clearly demonstrate that an electron-transfer quenching mechanism is operative and that I(2)(-)(*) can be photoproduced in high yield, phi = 0.25. For Ru(bpy)(3)(PF(6))(2) in acetonitrile, similar behavior could be observed at iodide concentrations >100 times that required for dichloromethane. PMID- 15877417 TI - A 3-D noninterpenetrating diamondoid network of a decanuclear copper(I) complex. AB - The solvothermal reaction of CuCl(2).2H(2)O with pyridine-4-thiol and ethanol yielded a novel photoluminescent 3-D polymeric complex with an interesting decorated diamondoid network that is constructed of decanuclear copper(I) cluster units and mu(4)-Cl ligands. The in situ generation of CuSO(4).5H(2)O implies the spontaneous occurrence of desulfurization and redox reactions in the present system. PMID- 15877418 TI - Magnetic polyoxometalates: anisotropic exchange interactions in the moiety of [(NaOH2)Co3(H2O)(P2W15O56)2]17-. AB - The magnetic exchange interactions in a C0(3)(11) moiety encapsulated in Na(17) [(NaOH(2))Co(3)(H(2)O)(P(2)W(15)O(56))(2)] (NaCo(3)) were studied by a combination of magnetic measurements (magnetic susceptibility and low-temperature magnetization), with a detailed Inelastic Neutron Scattering (INS) investigation. The novel structure of the salt was determined by X-ray crystallography. The ferromagnetic Co(3)O(14) triangular cluster core consists of three octahedrally oxo-coordinated Co(II) ions sharing edges. According to the single-ion anisotropy and spin-orbit coupling usually assumed for octahedral Co(II) ions, the appropiate exchange Hamiltonian to describe the ground-state properties of the isosceles triangular Co(3) spin cluster is anisotropic and is expressed as H = - 2sigma(alpha)(=)(x,y,z)(J(alpha)(12)S(1alpha)S(2alpha) + J(alpha)(23)S(2alpha)S(3alpha) + J(alpha)(13)S(1alpha)S(3alpha)), where J(alpha) are the components of the exchange interactions between the Co(II) ions. To reproduce the INS data, nonparallel anisotropic exchange tensors needed to be introduced, which were directly connected to the molecular symmetry of the complex. The following range of parameters (value +/- 0.5 cm(-1)) was found to reproduce all experimental information while taking magnetostructural relations into account: J(x)(12) = J(y)(13) = 8.6 cm(-1); J(y)(12) = J(x)(13) = 1.4 cm(-1); J(z)(12) = J(z)(13) = 10.0 cm(-1); J(x)(23) = J(y)(23) = 6.5 cm(-1) and = 3.4 cm( 1). PMID- 15877419 TI - Electropolymerization of vinylbipyridine complexes of ruthenium(II) and osmium(II) in SiO2 sol-gel films. AB - PF(6)(-) salts of the complexes [Ru(vbpy)(3)](2+) and [Os(vbpy)(3)](2+) (vbpy = 4 methyl-4'-vinyl-2,2'-bipyridine) have been electropolymerized into the pores of SiO(2) sol-gel films deposited on conductive Tin(IV)-doped indium oxide-coated glass slides (ITO, In(2)O(3):Sn). The resulting transparent composites represent a new class of materials of general formulas ITO/SG-poly-[Ru(vbpy)(3)](PF(6))(2) and ITO/SG-poly-[Os(vbpy)(3)](PF(6))(2). The composites are stable with respect to loss of complexes to the external solution and demonstrate several interesting phenomena: (1) Sol-gel pores, serving as diffusion channels for the vbpy complexes and counterions, play a key role in the formation of the polymer and dictate the electrochemical properties of the resulting composite. (2) Dynamic polymer growth occurs within individual diffusion channels creating parallel structures of filled and unfilled channels. (3) Unidirectional charge transfer and a "bilayer" effect have been shown to operate in ITO/SG-poly [Ru(vbpy)(3)](PF(6))(2) films exposed to [Os(vbpy)(3)](PF(6))(2) in the external solution. (4) Photophysical properties of the metal-to-ligand charge transfer (MLCT) excited states in ITO/SG-poly-[Ru(vbpy)(3)](PF(6))(2) composites are significantly modified compared to electropolymerized films on ITO or model monomeric complexes in solution. PMID- 15877420 TI - Use of phosphorus ligand NMR probes to investigate electronic and second-sphere solvent effects in ligand substitution reactions at manganese(II) and manganese(III). AB - Manganese/ligand association dynamics were studied using a series of structurally related anionic phosphorus ester ligand probes [CH(3)OP(O)(X)(Y)(-), where X = CH(3)O, CH(3)CH(2), or H and Y = O, S, or BH(3)]. Reactions of the probe ions with Mn(H(2)O)(6)(2+) and a manganese(III) porphyrin (Mn(III)TMPyP(5+)) were studied in aqueous solution by paramagnetic (31)P NMR line-broadening techniques. A satisfactory linear free energy relationship for reactions of the probe ions with Mn(H(2)O)(6)(2+) and Mn(III)TMPyP(5+) required consideration of both the basicity and solvent affinity of the probe ligands: log(k(app)) = log(k(0)) + alpha pK(a) + beta log(K(ext)), where k(0), alpha, and beta are metal complex dependent parameters and pK(a) and K(ext) represent the measured Bronsted acidity and water/n-butanol extraction constant for the probe anions, respectively. Reactions of Mn(H(2)O)(6)(2+) were relatively insensitive to changes in ligand basicity (alpha = -0.04) and favored the more hydrophilic anions (beta = -0.54). These observations are consistent with a dissociative ligand exchange mechanism wherein the outer-sphere complex is stabilized by hydrogen bonding between Mn(H(2)O)(6)(2+) and the incoming ligand. In contrast, reactions with Mn(III)TMPyP(5+) are accelerated by decreases in both the basicity (alpha = 0.43) and the hydrophilicity (beta = +0.97) of the probe. We conclude that reactions of Mn(III)TMPyP(5+) are also dissociative but that the aromatic groups of the porphyrin provide a hydrophobic environment surrounding the ligand binding site in Mn(III)TMPyP(5+). Thus, the probe/water solvent interactions must be significantly weakened in order to form the outer-sphere complex that leads to ligand substitution. This work demonstrates the utility of phosphorus relaxation enhancement (PhoRE) techniques for characterizing the second coordination sphere environment of metal complexes leading to ligation and will allow comparison of the second coordination spheres of Mn(H(2)O)(6)(2+) and Mn(III)TMPyP(5+) to those of other metal complexes. PMID- 15877421 TI - Influence of a gold(I)-Acetylide subunit on the photophysics of Re(phen)(CO)3Cl. AB - The synthesis and photophysical properties of two new Re(I) complexes are reported: fac-Re(phenC triple-bond CH)(CO)(3)Cl (where phenC triple bond CH is 5 ethynyl-1,10-phenanthroline) and its Au(I)-acetylide analogue (fac-Re(phenC triple-bond CAuPPh(3))(CO)(3)Cl). Also reported are the photophysical measurements obtained for the benchmark fac-Re(phen)(CO)(3)Cl chromophore, as well as the phenC triple-bond CAuPPh(3) and phenC triple-bond CH ligands. The unstable nature of the precursor gold-containing ligand illustrates the advantage of using the "chemistry on the complex" approach, which facilitated preparation of the Re-Au binuclear complex. Where possible, all compounds were studied by static and transient absorption (TA), as well as steady-state and time-resolved photoluminescence (TRPL), at room temperature (RT) and 77 K, as well as nanosecond time-resolved infrared (TRIR) spectroscopy. The spectroscopic information provided by these techniques enabled a thorough evaluation of excited state decay in most cases. In fac-Re(phenC triple bond CH)(CO)(3)Cl, the RT excited-state decay is most consistent with a metal-to-ligand charge transfer (MLCT) assignment, whereas at 77 K, the lowest excited state is dominated by the triplet intraligand ((3)IL) state, localized within the diimine ligand. The lowest excited state in fac-Re(phenC triple-bond CAuPPh(3))(CO)(3)Cl seems to result from an admixture of Re-based MLCT and (3)IL states resident on the phenC triple-bond CAuPPh(3) moiety. TA and TRIR methods indicate that these excited states are thermally equilibrated at room temperature. At 77 K, the MLCT energy of fac-Re(phenC triple-bond CAuPPh(3))(CO)(3)Cl is increased as a result of the glassy medium and the resulting excited state can be considered to be ligand localized. PMID- 15877422 TI - An asymmetric dizinc phosphodiesterase model with phenolate and carboxylate bridges. AB - A phosphodiesterase model with two zinc centers has been synthesized and characterized. The compound, [Zn(2)(L(-)(2H))(AcO)(H(2)O)](PF(6)).2H(2)O (Zn(2)L'), was formed using an "end-off" type compartmental ligand, 2,6-bis{[(2 pyridylmethyl)(2-hydroxyethyl)amino]methyl}-4-methylphenol (L), and zinc acetate dihydrate. The X-ray crystallographic analysis shows that Zn(2)L' contains a mu acetato-mu-cresolato-dizinc(II) core comprised of a quasi-trigonal bipyramidal Zn and a distorted octahedral Zn, and the distance between them is 3.421 Angstroms which is close to the dizinc distance in related natural metalloenzymes. Phosphodiesterase activity of Zn(2)L' was investigated using bis(4-nitrophenyl) phosphate (BNPP) as the substrate. The pH dependence of the BNPP cleavage in aqueous buffer media shows a sigmoid-shaped pH-k(obs) profile with an inflection point around pH 7.13 which is close to the first pK(a) value of 7.20 for Zn(2)L' obtained from the potentiometric titration. The catalytic rate constant (k(cat)) is 4.60 x 10(-6) s(-1) at pH 7.20 and 50 degrees C which is ca. 10(5)-fold higher than that of the uncatalyzed reaction. The deprotonated alcoholic group appended on Zn(2)L' is responsible for the cleavage reaction. The possible mechanism for the BNPP cleavage promoted by Zn(2)L' is proposed on the basis of kinetic and spectral analysis. The dizinc complex formed in situ in anhydrous DMSO exhibits a similar ability to cleave BNPP. This study provides a less common example for the phosphodiesterase model in which the metal-bound alkoxide is the nucleophile. PMID- 15877423 TI - Titanium(IV) citrate speciation and structure under environmentally and biologically relevant conditions. AB - The water-soluble complexes of Ti(IV) with citrate are of interest in environmental, biological, and materials chemistry. The aqueous solution speciation is revealed by spectropotentiometric titration. From pH 3-8, given at least three equivalents of ligand, 3:1 citrate/titanium complexes predominate in solution with successive deprotonation of dangling carboxylates as the pH increases. In this range and under these conditions, hydroxo- or oxo-metal species are not supported by the data. At ligand/metal ratios between 1:1 and 3:1, the data are difficult to fit, and are consistent with the formation of such hydroxo- or oxo- species. Stability constants for observed species are tabulated, featuring log beta-values of 9.18 for the 1:1 complex [Ti(Hcit)](+), and 16.99, 20.41, 16.11, and 4.07 for the 3:1 complexes [Ti(H(2)cit)(3)](2-), [Ti(H(2)cit)(Hcit)(2)](4-), [Ti(Hcit)(2)(cit)](6-), and [Ti(cit)(3)](8-), respectively (citric acid = H(4)cit). Optical spectra for the species are reported. The complexes exhibit similar yet distinct spectra, featuring putative citrate-to-Ti(IV) charge-transfer absorptions (lambda(max) approximately 250-310 nm with epsilon approximately 5000-7000 M(-)(1) cm(-1)). The prevailing 3:1 citrate/titanium ratio in solution is supported by electrospray mass spectrometry data. The X-ray crystal structure of a fully deprotonated tris-citrate complex Na(8)[Ti(C(6)H(4)O(7))(3)].17H(2)O (1) (or Na(8)[Ti(cit)(3)].17H(2)O) that crystallizes from aqueous solution at pH 7-8 is reported. Compound 1 crystallizes in the triclinic space group P, with a = 11.634(2) Angstroms, b = 13.223(3) Angstroms, c = 13.291(3) Angstroms, V = 1982.9(7) Angstroms(3), and Z = 2. PMID- 15877424 TI - Synthesis and structural characterization of some selenoruthenates and telluroruthenates. AB - The reaction of solid [RuClCp(PPh(3))(2)] with TeSe(3)(2-) or Se(n)(2-) in DMF leads to the formation of [RuCp(PPh(3))(mu(2)-Se(2))](2) (1). In the structure of this compound the two bridging Se(2) groups lead to a six-membered Ru(2)Se(4) ring in a chair conformation. Attached to each Ru center is a PPh(3) ligand in an equatorial position and a Cp ring in an axial position. The compound is diamagnetic. The compound [Ru(2)Cp(2)(mu(3)-Se(2))(mu(3)-Se)](2) (2) is obtained under similar conditions in the presence of air. This structure comprises a centrosymmetric Ru(4)Se(6) dimer formed from the two bridging Se groups and the two bridging Se(2) groups. Each Ru center is pi-bonded to a Cp ring. The reaction of solid [RuClCp(PPh(3))(2)] with a Te(n)(2-) polytelluride solution in DMF leads to the diamagnetic compound [(RuCp(PPh(3)))(2)(mu(2)-(1,4-eta:3,6-eta)Te(6))] (3). Here the Ru centers are bound to a bridging Te(6) chain at the 1, 4, 3, and 6 positions, leading to a bicyclic Ru(2)Te(6) ring. Each Ru atom is bound to a Cp ring and a PPh(3) group. This dimer possesses a center of symmetry. The structure of 3 is the first example of a bicyclic complex where fusion occurs along a Te-Te bond. If the same reaction is carried out in DMF/CH(2)Cl(2), rather than DMF, then [(RuCp(PPh(3)))(2)(mu(2)-(1,4-eta:3,6-eta)Te(6))].CH(2)Cl(2) (4) is obtained. In the solid state it possesses the same Ru(2)Te(6) structural unit as does 3, but the unit lacks a crystallographically imposed center of symmetry. The electronic structures of 3 and 4 have been analyzed with the use of first principles density functional theory. Bond order analysis indicates that the Te Te bond where fusion occurs has a shared bonding charge of about (2)/(3) of that found for Te-Te single bonds. PMID- 15877425 TI - Silylchalcogenolates MESiR(t)Bu(2) (M = Na, Cu, Zn, Fe; E = S, Se, Te; R = tBu, Ph) and disilyldichalcogenides tBu2RSiE-ESiRtBu2 (E = S, Se, Te; R = tBu, Ph): synthesis, properties, and structures. AB - The sodium silyl chalcogenolates NaESiR(t)Bu(2) (R = Ph, (t)Bu; E = S, Se, Te), accessible by the nucleophilic degradation of S, Se, or Te by the sodium silanides NaSiR(t)Bu(2) (R = Ph, (t)Bu), have been characterized by X-ray structure analysis. Protonolysis of the sodium silyl chalcogenolates yields the corresponding chalcogenols. The Cu and Zn chalcogenolates, [Cu(SSiPh(t)Bu(2))](4) and [ZnCl(SSi(t)Bu(3))(THF)](2), have been synthesized by metathesis reactions of CuCl with NaSSiPh(t)Bu(2) and of ZnCl(2) with NaSSi(t)Bu(3), respectively. The solid-state structures of the transition metal thiolates have been determined. The compounds (t)Bu(2)RSiE-ESiR(t)Bu(2) (R = Ph, (t)Bu; E = S, Se, Te) are accessible via air oxidation. With the exception of (t)Bu(3)SiS-SSi(t)Bu(3), these compounds were analyzed using X-ray crystallography and represent the first structurally characterized silylated heavy dichalcogenides. Oxidative addition of (t)Bu(3)SiTe-TeSi(t)Bu(3) to Fe(CO)(5) yields [Fe(TeSi(t)Bu(3))(CO)(3)](2), which has also been structurally characterized. PMID- 15877426 TI - Multielectron donors based on TTF-phosphine and ferrocene-phosphine hybrid complexes of a hexarhenium(III) octahedral cluster core. AB - Electroactive molecular materials precursors are obtained through coordination chemistry of the hexarhenium cluster core [Re(6)Se(8)](2+) on the six available apical positions with redox-active phosphines bearing tetrathiafulvalene- or ferrocene-based moieties. Single-crystal X-ray diffraction study and electrospray mass spectrometry ascertain the synthesis of these hexasubstituted electroactive clusters, containing up to 12 redox active sites. Cyclic voltammetry experiments demonstrate that these compounds can be reversibly oxidized at rather low potentials, thus allowing an easy access to the corresponding radical species which should provide new conducting and/or magnetic molecular materials. PMID- 15877427 TI - Metal-metal interactions in platinum(II)/gold(I) or platinum(II)/silver(I) salts containing planar cations and linear anions. AB - The salts [Pt{C(NHMe)(2)}(4)][Au(CN)(2)](2), [Pt{C(NHMe)(2)}(4)][Ag(2)(CN)(3)][Ag(CN)(2)], [Pt(en)(2)][Au(CN)(2)](2), [Pt(en)(2)][Ag(CN)(2)](2), and [Pt(bipy)(2)][Au(CN)(2)](2) have been prepared by mixing solutions of salts containing the appropriate cation with solutions of K[Au(CN)(2)] or K[Ag(CN)(2)]. Because the platinum atom in the cation is sterically protected, the structures of [Pt{C(NHMe)(2)}(4)][Au(CN)(2)](2) and [Pt{C(NHMe)(2)}(4)][Ag(2)(CN)(3)][Ag(CN)(2)] reveal no close metal-metal interactions. Colorless crystals of [Pt(en)(2)][Au(CN)(2)](2) and [Pt(en)(2)][Ag(CN)(2)](2) are isostructural and involve extended chains of alternating cations and anions that run parallel to the crystallographic a axis, along with isolated anions. In the chains, the metal-metal separations are relatively short: Pt...Au, 3.1799(3) Angstroms; Pt...Ag, 3.1949(2) Angstroms. In [Pt(bipy)(2)][Au(CN)(2)](2), each cation has axial interactions with the anions through close Pt...Au contacts [3.1735(6) Angstroms]. In addition, the anions are weakly linked through Au...Au contacts of 3.5978(9) Angstroms. Unlike the previously reported Pt/Au complex [Pt(NH(3))(4)][Au(CN)(2)](2).1.5H(2)O, which is luminescent, none of the salts reported here luminesce. PMID- 15877428 TI - Aerobic catechol oxidation catalyzed by a bis(mu-oxo)dimanganese(III,III) complex via a manganese(II)-semiquinonate complex. AB - A 3,5-di-tert-butyl-1,2-semiquinonato (DTBSQ) adduct of Mn(II) was prepared by a reaction between Mn(II)(TPA)Cl(2) (TPA = tris(pyridin-2-ylmethyl)amine) and DTBSQ anion and was isolated as a tetraphenylborate salt. The X-ray crystal structure revealed that the complex is formulated as a manganese(II)-semiquinonate complex [Mn(II)(TPA)(DTBSQ)](+) (1). The electronic spectra in solution also indicated the semiquinonate coordination to Mn. The exposure of 1 in acetonitrile to dioxygen afforded 3,5-di-tert-butyl-1,2-benzoquione and a bis(mu oxo)dimanganese(III,III) complex [Mn(III)(2)(mu-oxo)(2)(TPA)(2)](2+) (2). The reaction of 2 with 3,5-di-tert-butylcatechol (DTBCH(2)) quantitatively afforded two equivalents of 1 under anaerobic conditions. The highly efficient catalytic oxidation of DTBCH(2) with dioxygen was achieved by combining the above two reactions, that is, by constructing a catalytic cycle involving both manganese complexes 1 and 2. It was revealed that dioxygen is reduced to water but not to hydrogen peroxide in the catalytic cycle. PMID- 15877429 TI - Synthesis and properties of calcium alanate and two solvent adducts. AB - Several ways to synthesize solvated and desolvated calcium tetrahydroaluminate by wet-chemical and mechanochemical methods are presented. The products were characterized by elemental analysis, X-ray diffraction (XRD), and infrared spectroscopy (FTIR). The crystal structure of Ca(AlH(4))(2).4THF was determined. After desolvation, an ultrafine powder was obtained. IR data and the mass balance suggest a compound with the composition Ca(AlH(4))(2), containing tetrahedral [AlH(4)] groups. PMID- 15877430 TI - The reaction between ZnO and molten Na2S2O7 or K2S2O7 forming Na2Zn(SO4)2 orK2Zn(SO4)2, studied by Raman spectroscopy and x-ray diffraction. AB - Reactions between solid zinc oxide and molten sodium or potassium pyrosulfates at 500 degrees C are shown by Raman spectroscopy to be 1:1 reactions leading to solutions. By lowering the temperature of the solution melts, colorless crystals form. Raman spectra of the crystals are given and tentatively assigned. Crystal structures of the monoclinic salts at room temperature are given. Na(2)Zn(SO(4))(2): space group = P2/n (No. 13), Z = 8, a = 8.648(3) Angstroms, b = 10.323(3) Angstroms, c = 15.103(5) Angstroms, beta = 90.879(6) degrees, and wR(2) = 0.0945 for 2748 independent reflections. K(2)Zn(SO(4))(2): space group = P2(1)/n (No.14), Z = 4, a = 5.3582(11) Angstroms, b = 8.7653(18) Angstroms, c = 16.152(3) Angstroms, beta = 91.78(3) degrees , and wR(2) = 0.0758 for 1930 independent reflections. In both compounds, zinc is nearly perfectly trigonally bipyramidal, coordinated to five oxygen atoms, with Zn-O bond lengths ranging from 1.99 to 2.15 Angstroms, equatorial bonds being slightly shorter on the average. The O-Zn-O angles are approximately 90 degrees and 120 degrees . The sulfate groups connect adjacent Zn(2+) ions, forming complicated three dimensional networks. All oxygen atoms belong to nearly perfect tetrahedral SO(4)(2-) groups, bound to zinc. No oxygen atom is terminally bound to zinc; all zinc oxygens are further connected to sulfur atoms (Zn-O-S bridging). In both structures, some oxygen atoms are uniquely bound to certain S atoms. The sulfate group tetrahedra have quite short (1.42-1.45 Angstroms) terminal S-O bonds in comparison to the longer (1.46-1.50 Angstroms) Zn-bridging S-O bonds. The Na(+) or K(+) ions adopt positions between the ZnO(5) hexahedra and the SO(4) tetrahedra, completing the three-dimensional network of the M(2)Zn(SO(4))(2) structures. Bond distances and angles compare well with literature values. Empirical correlations between S-O bond distances and average O-S-O bond angles follow a previously found trend. PMID- 15877431 TI - Synthesis and characterization of a twin Cubane-type molybdenum-rhodium-sulfur cluster, [{Mo3RhCp*S4(H2O)7(O)}2]8+. X-ray structure of [{Mo3RhCpS4(H2O7O}2](CH3C6H4SO3)(8).14H2O. AB - The reaction of [Mo(3)S(4)(H(2)O)(9)](4+) (1) with [(CpRhCl(2))(2)] afforded a novel rhodium-molybdenum cluster, [{Mo(3)RhCpS(4)(H(2)O)(7)(O)}(2)](8+) (2). X ray structure analysis of [2](pts)(8).14H(2)O (pts(-) = CH(3)C(6)H(4)SO(3)(-)) has revealed the existence of a new oxo-bridged twin cubane-type core, (Mo(3)RhCpS(4))(2)(O)(2). The high affinity of the CpRh group for sulfur atoms in 1 seems to be the main driving force for this reaction. The strong Lewis acidity of the CpRh group in intermediate A, [Mo(3)RhCpS(4)(H(2)O)(9)](6+), caused a release of proton from one of the water molecules attached to the molybdenum atoms to give intermediate B, [Mo(3)RhCpS(4)(H(2)O)(8)(OH)](5+). The elimination of two water molecules from two intermediate B molecules, followed by the deprotonation reaction of hydroxo bridges, generated the twin cubane-type cluster 2. The formal oxidation states of rhodium and molybdenum atoms are the same before and after the reaction (i.e., Mo(IV)(3), Rh(III)). The Mo-O-Mo moieties in [2](pts)(8).14H(2)O are nearly linear with a bond angle of 164.3(3) degrees, and the basicity of the bridging oxygen atoms seems to be weak. For this reason, protonation at the bridging oxygen atoms does not occur even in a strongly acidic aqueous solution. The binding energy values of Mo 3d(5/2), Rh 3d(5/2), and C 1s obtained from X-ray photoelectron spectroscopy measurements for [2](pts)(8).14H(2)O are 229.8, 309.3, and 285 eV, respectively. The XPS measurements on the Rh 3d(5/2) binding energy indicate that the oxidation state of Rh is 3+. The binding energy of Mo 3d(5/2) (229.8 eV) compares with that observed for [1](pts)(4).7H(2)O (230.7 eV, Mo 3d(5/2)). A lower energy shift (0.9 eV) is observed in the binding energy of Mo 3d(5/2) for [2](pts)(8).14H(2)O. This energy shift may correspond to the coordination of an oxygen atom having a negative charge to the molybdenum atom. PMID- 15877432 TI - Effect of 2-(2-Pyridyl)azole-based ancillary ligands (L1-4) on the electrophilicity of the nitrosyl function in [RuII(trpy)(L1-)4)(NO)]3+ [trpy = 2,2':6',2' '-Terpyridine]. synthesis, structures, and spectroscopic, electrochemical, and kinetic aspects. AB - Ruthenium nitrosyl complexes [Ru(trpy)(L(1-4))(NO)](3+) (13-16) [trpy = 2,2':6',2' '-terpyridine, L(1) = 2-(2-pyridyl)benzoxazole, L(2) = 2-(2 pyridyl)benzthiazole, L(3) = 2-(2-pyridyl)benzimidazole, L(4) = 1-methyl-2-(2 pyridyl)-1H-benzimidazole] were obtained in a stepwise manner starting from [Ru(II)(trpy)(L(1-4))(Cl)]ClO(4) (1-4) -->[Ru(II)(trpy)(L(1 4))(H(2)O)](ClO(4))(2) (5-8) --> [Ru(II)(trpy)(L(1-4)) (NO(2))]ClO(4) (9-12) --> [Ru(II)(trpy)(L(1,2,4))(NO)](ClO(4))(3) (13, 14, 16)/[Ru(II)(trpy)(L(3))(NO)](ClO(4))(2)(NO(3)) (15). Crystal structures of 1, 2, 4, 9, 12, 13, 15, and 16 established the stereoretentive nature of the transformation processes. Though the complexes of L(1), L(3), and L(4) were isolated in the isomeric form A (pi-acceptor trpy and azole ring in the equatorial plane and the pyridine and chloride donors in the axial positions), complexes of L(2) preferentially stabilized in form B (trpy and pyridine in the equatorial plane and the azole ring and chloride donors in the axial positions). The nu(NO) stretching frequency varied in the range of 1957-1932 cm(-1), 13 >> 14 approximately 15 > 16, primarily depending on the electronic aspects of L as well as the isomeric structural forms. The coordinated nitrosyl function underwent successive reductions of [Ru(II)-NO(+)](3+) --> [Ru(II)-NO(*)](2+) and [Ru(II) NO(*)](2+) --> [Ru(II)-NO(-)](+), and the first reduction potential follows the order 14 > 13 >> 15 approximately 16. The nearly axial EPR spectra having nitrogen hyperfine splittings (A approximately 26 G) at 77 K of 13(-)-16(-) with g approximately 2.0 established that the reduction process is largely centered around the nitrosyl function. Despite an appreciably high nu(NO), the complexes were found to be unusually stable even in the aqueous medium. They transformed slowly and only partially into the corresponding nitro derivatives in H(2)O (k approximately 10(-4) s(-1) and K = 0.4-3.8). The chloro (1-4), aqua (5-8), and nitro (9-12) derivatives displayed reasonably strong emissions near 700 nm at 77 K (phi = 10(-1)-10(-2)). The aqua derivative 7 was found to interact with the calf thymus and the circular form of p-Bluescript SK DNA. PMID- 15877433 TI - New hydrothermal synthesis and structure of Th2(PO4)2(HPO4).H2O: the first structurally characterized thorium Hydrogenphosphate. AB - Th(2)(PO(4))(2)(HPO(4)).H(2)O was synthesized under wet hydrothermal conditions starting from a mixture of H(3)PO(3) and Th(NO(3))(4).5H(2)O. The crystal structure was solved by powder X-ray diffraction data. The unit cell parameters are a = 6.7023(8) Angstroms, b = 7.0150(8) Angstroms, c = 11.184(1) Angstroms, beta = 107.242(4) degrees, space group P2(1), and Z = 2. The structure consists of layers of both thorium atoms and PO(4) groups, alternating with a layer formed by HPO(4) entities and water molecules. By thermal treatment, this compound turns into Th(4)(PO(4))(4)P(2)O(7), a ceramic already described in the field of the immobilization of tetravalent actinides. PMID- 15877434 TI - Zinc-thiolate complexes of the bis(pyrazolyl)(thioimidazolyl)hydroborate tripods for the modeling of thiolate alkylating enzymes. AB - The new tripod ligands bis(pyrazolyl)(3-tert-butyl-2-thioimidazol-1 yl)hydroborate (L(1)) and bis(pyrazolyl)(3-isopropyl-2-thioimidazol-1 yl)hydroborate (L(2)), together with zinc nitrate or zinc chloride and the corresponding thiolates, have yielded a total of 17 zinc-thiolate complexes. These comprise aliphatic as well as aromatic thiolates and a cysteine derivative. Structure determinations have confirmed the tetrahedral ZnN(2)S(2) coordination in the complexes. Upon reaction with methyl iodide, the species L(1).Zn-SR are slowly converted to L(1).Zn-I and the free thioethers CH(3)SR. A kinetic analysis has shown these alkylations to be about 1 order of magnitude slower than those of the tris(pyrazolyl)borate complexes Tp(Ph,Me)Zn-SR. Alkylations with trimethyl phosphate were found to proceed very slowly even in DMSO at 80 degrees C. PMID- 15877435 TI - Syntheses, structures, and magnetic properties of diphenoxo-bridged M(II)Ln(III) complexes derived from N,N'-ethylenebis(3-ethoxysalicylaldiimine) (M = Cu or Ni; Ln = Ce-Yb): observation of surprisingly strong exchange interactions. AB - A series of heterodinuclear Cu(II)Ln(III) and Ni(II)Ln(III) complexes, [M(II)L(1)Ln(III)(NO(3))(3)] (M = Cu or Ni; Ln = Ce-Yb), with the hexadentate Schiff base compartmental ligand N,N'-ethylenebis(3-ethoxysalicylaldiimine) (H(2)L(1)) have been synthesized and characterized. The X-ray crystal structure determinations of 13 of these compounds reveal that they are all isostructural. All of these complexes crystallize with the same orthorhombic P2(1)2(1)2(1) space group with closely similar unit cell parameters. Typically, the structure consists of a diphenoxo-bridged 3d-4f dinuclear core, self-assembled to two dimensions due to the intermolecular nitrate...copper(II) or nitrate...nickel(II) semicoordination and weak C-H...O hydrogen bonds. Despite that, the metal centers of the neighboring units are well separated (the ranges of the shortest intermolecular contacts (A) are (M...M) 7.46-7.60, (Ln...Ln) 8.56-8.69, and (M...Ln) 6.12-6.20). Variable-temperature (5-300 K) magnetic susceptibility measurements of all the complexes have been made. The nature of exchange interactions in the Cu(II)Ln(III) systems has been inferred from the Deltachi(M)T versus T plots, where Deltachi(M)T is the difference between the values of chi(M)T for a Cu(II)Ln(III) system and its corresponding Ni(II)Ln(III) analogue. Ferromagnetic interactions seem to be exhibited by the Cu(II)Gd(III), Cu(II)Tb(III), Cu(II)Dy(III), Cu(II)Ho(III), Cu(II)Tm(III), and Cu(II)Yb(III) complexes, while, for the Cu(II)Er(III) complex, no definite conclusion could be reached. On the other hand, among the lower members of the series, the complexes of Ce(III), Nd(III), and Sm(III) exhibit antiferromagnetic interactions, while the Cu(II)Pr(III) and Cu(II)Eu(III) analogues behave as spin-uncorrelated systems. The observations made here vindicate the proposition of Kahn (Inorg. Chem. 1997, 36, 930). The Deltachi(M)T versus T plots also suggest that, for most of the Cu(II)Ln(III) complexes, the exchange interactions are fairly strong, which probably could be related to the small dihedral angle (ca. 4 degrees) between the CuO(2) and LnO(2) planes. PMID- 15877436 TI - OH functionality of germanium(II) compounds for the formation of heterobimetallic oxides. AB - Two novel germanium(II) mu-oxo heterobimetallic oxides with different oxidation states at the metal centers have been reported. The reaction of LGeOH [L = N(Ar)C(Me)CHC(Me)N(Ar) (Ar = 2,6-i-Pr(2)C(6)H(3))] with Cp(2)MMe(2) (M = Zr, Hf) in Et(2)O afforded LGeOM(Me)Cp(2) [M = Zr (2), Hf (3)] in moderate yield. Compounds 2 and 3 were characterized by elemental analysis, IR, NMR, EI-MS, and single X-ray structural analysis. Compounds 2 and 3 crystallized in the space group P, and the geometry at the metal centers is tetrahedral. The Ge-O bond lengths of 2 and 3 are very similar (1.797(2) and 1.799(3) Angstroms, respectively), and a bent M-O-M' angle in 2 (143.8(1) degrees) and 3 (141.9(2) degrees) features both oxide systems. Different orientations of the Cp and Me groups of the metal centers were observed, and deviations of the Cp groups were exhibited. PMID- 15877437 TI - Catalytic and direct oxidation of cysteine by octacyanomolybdate(V). AB - The oxidation of cysteine by [Mo(CN)(8)](3-) in deoxygenated aqueous solution at a moderate pH is strongly catalyzed by Cu(2+), to the degree that impurity levels of Cu(2+) are sufficient to dominate the reaction. Dipicolinic acid (dipic) is a very effective inhibitor of this catalysis, such that with 1 mM dipic, the direct oxidation can be studied. UV-vis spectra and electrochemistry show that [Mo(CN)(8)](4-) is the Mo-containing product. Cystine and cysteinesulfinate are the predominant cysteine oxidation products. The stoichiometric ratio (Deltan(Mo(V))/Deltan(cysteine)) of 1.4 at pH 10.8 is consistent with this product distribution. At pH 1.5, the reaction is quite slow and yields intractable kinetics. At pH 4.5, the rates are much faster and deviate only slightly from pseudo-first-order behavior. With 2 mM PBN (N-phenyl-tert-butyl nitrone) present at pH 4.5, the reaction rate is about 20% less and shows excellent pseudo-first-order behavior, but the stoichiometric ratio is not significantly changed. The rates also display a significant specific cation effect. In the presence of spin-trap PBN, the kinetics were studied over the pH range 3.48-12.28, with [Na(+)] maintained at 0.09-0.10 M. The rate law is d[Mo(V)]/dt = k[cysteine](tot)[Mo(V)], with k = {2(k(b)K(a1)K(a2)[H(+)] + k(c)K(a1)K(a2)K(a3))}/([H(+)](3) + K(a1)[H(+)](2) + K(a1)K(a2)[H(+)] + K(a1)K(a2)K(a3)), where K(a1), K(a2), and K(a3) are the successive acid dissociation constants of HSCH(2)CH(NH(3)(+))CO(2)H. Least-squares fitting yields k(b) = (7.1 +/- 0.4) x 10(4) M(-1) s(-1) and k(c) = (2.3 +/-0.2) x 10(4) M(-1) s( 1) at mu = 0.1 M (NaCF(3)SO(3)) and 25 degrees C. A mechanism is inferred in which k(b) and k(c) correspond to electron transfer to Mo(V) from the thiolate forms of anionic and dianionic cysteine. PMID- 15877438 TI - Complexes of functionalized dipyrido[3,2-a:2',3'-c]-phenazine: a synthetic, spectroscopic, structural, and density functional theory study. AB - The ligands 11-bromodipyrido[3,2-a:2',3'-c]phenazine and ethyl dipyrido[3,2 a:2',3'-c]phenazine-11-carboxylate have been prepared and coordinated to ruthenium(II), rhenium(I), and copper(I) metal centers. The electronic effects of substitution of dipyrido[2,3-a:3',2'-c]phenazine (dppz) have been investigated by spectroscopy and electrochemistry, and some photophysical properties have been studied. The crystal structures of [Re(L)(CO)(3)Cl] (L = ethyl dipyrido[3,2 a:2',3'-c]phenazine-11-carboxylate or 11-bromodipyrido[3,2-a:2',3'-c]phenazine) are presented. Density functional theory calculations on the complexes show only small deviations in bond lengths and angles (most bonds within 0.02 Angstroms, most angles within 2 degrees) from the crystallographic data. Furthermore, the vibrational spectra of the strongest Raman and IR bands are predicted to within an average 6 cm(-1) for the complexes [Re(L)(CO)(3)Cl] and [Cu(L)(triphenylphosphine)(2)]BF(4) (in the 1000-1700 cm(-1) region). Spectroscopic and electrochemical evidence suggest that reduction of the complex causes structural changes across the entire dppz ligand. This is unusual as dppz based ligands typically have electrochemical properties that suggest charge localization with reduction on the phenazine portion of the ligand. The excited state lifetimes of the complexes have been measured, and they range from ca. 200 ns for the [Ru(L)(2,2'-bipyridine)(2)](PF(6))(2) complexes to over 2 mus for [Cu(11-bromodipyrido[3,2-a:2',3'-c]phenazine)(PPh(3))(2)](BF(4)) at room temperature. The emission spectra suggest that the unusually long-lived excited states of the copper complexes result from metal-to-ligand charge transfer (MLCT) transitions as they are completely quenched in methanol. Electroluminescent films may be fabricated from these compounds; they show MLCT state emission even at low doping levels [<0.1% by weight in poly(vinylcarbazole) polymer matrix]. PMID- 15877439 TI - Separation and characterization of the two diastereomers for [Gd(DTPA-bz NH2)(H2O)]2-, a common synthon in macromolecular MRI contrast agents: their water exchange and isomerization kinetics. AB - Chiral, bifunctional poly(amino carboxylate) ligands are commonly used for the synthesis of macromolecular, Gd(III)-based MRI contrast agents, prepared in the objective of increasing relaxivity or delivering the paramagnetic Gd(III) to a specific site (targeting). Complex formation with such ligands results in two diastereomeric forms for the complex which can be separated by HPLC. We demonstrated that the diastereomer ratio for Ln(III) DTPA derivatives (approximately 60:40) remains constant throughout the lanthanide series, in contrast to Ln(III) EPTPA derivatives, where it varies as a function of the cation size with a maximum for the middle lanthanides (DTPA(5-) = diethylenetriaminepentaacetate; EPTPA(5-) = ethylenepropylenetriaminepentaacetate). The interconversion of the two diastereomers, studied by HPLC, is a proton-catalyzed process (k(obs) = k(1)[H(+)]). It is relatively fast for [Gd(EPTPA-bz-NH(2))(H(2)O)](2-) but slow enough for [Gd(DTPA-bz-NH(2))(H(2)O)](2-) to allow investigation of pure individual isomers (isomerization rate constants are k(1) = (3.03 +/- 0.07) x 10(4) and 11.6 +/- 0.5 s(-1) M(-1) for [Gd(EPTPA-bz-NH(2))(H(2)O)](2)(-) and [Gd(DTPA-bz-NH(2))(H(2)O)](2-), respectively). Individual water exchange rates have been determined for both diastereomers of [Gd(DTPA-bz-NH(2))(H(2)O)](2-) by a variable-temperature (17)O NMR study. Similarly to Ln(III) EPTPA derivatives, k(ex) values differ by a factor of 2 (k(ex)(298) = (5.7 +/- 0.2) x 10(6) and (3.1 +/- 0.1) x 10(6) s(-1)). This variance in the exchange rate has no consequence on the proton relaxivity of the two diastereomers, since it is solely limited by fast rotation. However, such difference in k(ex) will affect proton relaxivity when these diastereomers are linked to a slowly rotating macromolecule. Once the rotation is optimized, slow water exchange will limit relaxivity; thus, a factor of 2 in the exchange rate can lead to a remarkably different relaxivity for the diastereomer complexes. These results have implications for future development of Gd(III)-based, macromolecular MRI contrast agents, since the use of chiral bifunctional ligands in their synthesis inevitably generates diastereomeric complexes. PMID- 15877440 TI - Influence of steric and electronic properties of the defect site, lanthanide ionic radii, and solution conditions on the composition of lanthanide(III) alpha1 P2W17O6110- polyoxometalates. AB - This study identifies the principles that govern the formation and stability of Ln complexes of the (alpha(1)-P(2)W(17)O(61))(10-) isomer. The conditional stability constants for the stepwise formation equilibria, K(1cond) and K(2cond), determined by (31)P NMR spectroscopy, show that the high log K(1cond)/log K(2cond) ratio predicts the stabilization of the 1:1 Ln/ (alpha(1) P(2)W(17)O(61))(10-) species. The value of log K(1cond) increases as the Ln series is traversed, consistent with the high charge/size requirement of the basic alpha(1) defect site. The conditional stability constants, K(2), are very low and are highly dependent on the countercations in the buffer. The source of the instability is understood from the crystal structures of the early-mid lanthanide analogues, where the close contact of the (alpha(1)-P(2)W(17)O(61))(10 ) units result in severe steric encumbrance. The electronic properties of the alpha(1) defect along with the lanthanide ionic radii and countercation composition are important parameters that need to be considered for a rational synthesis of lanthanide polyoxometalates. PMID- 15877441 TI - Density functional theory study of 11-atom germanium clusters: effect of electron count on cluster geometry. AB - Density functional theory (DFT) at the hybrid B3LYP level has been applied to the germanium clusters Ge(11)(z) (z = -6, -4, -2, 0, +2, +4, +6) starting from eight different initial configurations. The global minimum within the Ge(11)(2-) set is an elongated pentacapped trigonal prism distorted from D(3)(h) to C(2v) symmetry. However, the much more spherical edge-coalesced icosahedron, also of C(2v) symmetry, expected by the Wade-Mingos rules for a 2n + 2 skeletal electron system and found experimentally in B(11)H(11)(2-) and isoelectronic carboranes, is of only slightly higher energy (+5.2 kcal/mol). Even more elongated D(3)(h) pentacapped trigonal prisms are the global minima for the electron-rich structures Ge(11)(4-) and Ge(11)(6-). For Ge(11)(4-) the C(5v) 5-capped pentagonal antiprism analogous to the dicarbollide ligand C(2)B(9)H(11)(2-) is of significantly higher energy (approximately 28 kcal/mol) than the D(3h) global minimum. The C(2v) edge-coalesced icosahedron is also the global minimum for the electron-poor Ge(11) similar to its occurrence in experimentally known 11-vertex "isocloso" metallaboranes of the type (eta(6)-arene)RuB(10)H(10). The lowest energy polyhedral structures computed for the more hypoelectronic Ge(11)(4+) and Ge(11)(6+) clusters are very similar to those found experimentally for the isoelectronic ions E(11)(7-) (E = Ga, In, Tl) and Tl(9)Au(2)(9-) in intermetallics in the case of Ge(11)(4+) and Ge(11)(6+), respectively. These DFT studies predict an interesting D(5h) centered pentagonal prismatic structure for Ge(11)(2+) and isoelectronic metal clusters. PMID- 15877442 TI - Analysis of the refractive indices of TiO2, TiOF2, and TiF4: concept of optical channel as a guide to understand and design optical materials. AB - In an effort to gain insight into how optical properties of insulating materials are affected by a change in chemical composition, we investigated the dielectric functions of titanium dioxide, TiO(2), and its fluorine-substituted phases, TiOF(2) and TiF(4), by electron energy loss spectroscopy measurements and density functional theory electronic band structure calculations. The refractive indices of these compounds are found to be inversely proportional to their cell volumes per formula unit. This observation was explained by employing the concept of optical channels. Our study indicates that the light-scattering properties of insulating compounds can be controlled by modifying their cell volumes. PMID- 15877443 TI - Structural and electronic factors controlling the refractive indices of the chalcogenides ZnQ and CdQ (Q = O, S, Se, Te). AB - On the basis of first-principles electronic band structure theory, we calculated the refractive indices of the zinc blende and wurtzite structures of the chalcogenides ZnQ and CdQ (Q = O, S, Se, Te) and analyzed their trends by calculating the total absorption power per unit formula. The calculated refractive indices are in good agreement with the available experimental data. The total absorption power per formula unit is found to allow one to distinguish the effect of the chromophore from that of the cell volume on the refractive indices of insulating inorganic compounds. PMID- 15877444 TI - Three-, two-, and one-dimensional metal phosphonates based on [hydroxy(4 pyridyl)methyl]phosphonate: M{(4-C5H4N)CH(OH)PO3}(H2O) (M = Ni, Cd) and Gd{(4 C5H4N)CH(OH)P(OH)O2}3.6H2O. AB - Based on the [hydroxy(4-pyridyl)methyl]phosphonate ligand, three compounds with formula Ni{(4-C(5)H(4)N)CH(OH)PO(3)}(H(2)O) (1), Cd{(4 C(5)H(4)N)CH(OH)PO(3)}(H(2)O) (2), and Gd{(4-C(5)H(4)N)CH(OH)P(OH)O(2)}(3).6H(2)O (3) have been synthesized under hydrothermal conditions. The crystal data for 1 are as follows: orthorhombic, space group Pbca, a = 8.7980(13) A, b = 10.1982(15) A, and c = 17.945(3) A. For 2 the crystal data are as follows: monoclinic, space group C2/c, a = 23.344(6) Angstroms, b = 5.2745(14) Angstroms, c = 16.571(4) Angstroms, and beta = 121.576(4) degrees. The crystal data for 3 are as follows: rhombohedral, space group R, a = 22.2714(16) Angstroms, b = 22.2714(16) Angstroms, and c = 9.8838(11) Angstroms. Compound 1 adopts a three-dimensional pillared layered structure in which the inorganic layers made up of corner sharing {NiO(5)N} octahedra and {CPO(3)} tetrahedra are connected by pyridyl groups. A two-dimensional layer structure is found in compound 2, which contains alternating inorganic double chains and pyridyl rings. Compound 3 has a one dimensional chain structure where the Gd atoms are triply bridged by O-P-O linkages. The pyridyl nitrogen atom in 3 remains uncoordinated and is involved in the interchain hydrogen bonds. Magnetic susceptibility studies of 1 and 3 reveal that weak ferromagnetic interactions are mediated between Ni(II) centers in compound 1. For compound 3, the behavior is principally paramagnetic. PMID- 15877445 TI - Spectroscopic and computational studies on [Ni(tmc)CH3]OTf: implications for Ni methyl bonding in the A cluster of acetyl-CoA synthase. AB - The five-coordinate high-spin (S = 1) Ni(2+) complex [Ni(tmc)CH(3)](+) (1) (tmc = 1,4,8,11-tetramethyl-1,4,8,11-tetraazacyclotetradecane) serves as a model for a viable reaction intermediate of the A cluster of acetyl-CoA synthase (ACS) in which the distal nickel center is methylated. Spectroscopic and density functional theory (DFT) computational studies afford a quantitative bonding description for 1 that reveals a highly covalent Ni-CH(3) bond. From a normal coordinate analysis of resonance Raman data obtained for 1, a value of k(Ni-C) = 1.44 mdyn/Angstroms is obtained for the Ni-C stretch force constant of this species. This value is smaller than k(Co)(-C) = 1.85 mdyn/Angstroms, which is reported for the Co-C stretch in the methylcobinamide cofactor (5) that serves as the methyl donor to the A cluster in the ACS catalytic cycle. Experimentally calibrated DFT computations on viable methylated A cluster models reveal that the methyl group binds to the proximal (Ni(p)) rather than the distal (Ni(d)) nickel center and afford a simple electronic argument for this preference. By correlating the experimental force constants with the computed bond orders of the M-C bonds in 1 and 5, the Ni(p)(2+)-CH(3) bond strength for an A cluster model with a square-planar Ni(p) conformation, which is the most probable structure of the methylated A cluster on the basis of steric and energetic considerations, is predicted to be similar to the Co(3+)-CH(3) bond strength in CH(3)-CoFeSP. This similarity could be a crucial thermodynamic prerequisite for the reversibility of the enzymatic transmethylation reaction. PMID- 15877446 TI - A second-order nonlinear optical material prepared through in situ hydrothermal ligand synthesis. AB - The in situ hydrothermal reactions of ZnCl(2) with benzonitrile, 2-amino-5 cyanopyridine, and trans-2,3-dihydro-2-(4' '-cyanophenyl)-benzo[e]indole in the presence of NaN(3) and water afford two 3D-diamond-like networks, (CN(4) C(6)H(5))(2)Zn (1) and (NH(2)-C(5)H(3)N-CN(4))(2)Zn (2), and one 2D square grid network, [(CN(4)-C(6)H(4)-C(12)H(7)N-C(5)H(4)N)(2)Zn].1.5H(2)O (3), in which these ligands gradually involve a noncenter-A-D (acceptor-donor) system, a one center-A-D system, and a two-center-A-D system, respectively. All three compounds crystallize in noncentrosymmetric space groups (I2d for 1 and 2 and Fdd2 for 3) and display strong second harmonic generation (SHG) responses. Among the three new complexes, 3 shows the largest SHG effect, which is about 50 and 500 times that of urea and KDP (KH(2)PO(4)), respectively. The two-center-A-D system (multicenter push-pull electronic effect) in 3 may be responsible for it having the largest SHG effect. Interestingly, the three compounds exhibit strong fluorescent emissions at different wavelengths, 1 and 2 with blue fluorescent emissions at 390 and 415 nm and 3 with yellow-green fluorescent emissions at 495 and 532 nm. PMID- 15877447 TI - Multinuclear Fe(III) complexes with polydentate ligands of the family of dicarboxyimidazoles: nuclearity- and topology-controlled syntheses and Magneto structural correlations. AB - Two polydentate ligands of the family of dicarboxyimidazoles, H(2)MeDCBI (= 4,5 dicarboxy-1-methyl-1H-imidazole) and H(3)DCBI (= 4,5-dicarboxyimidazole), have been used in reactions with the [Fe(3,5-(t)()Bu(2)salpn)](+) species {3,5 (t)Bu(2)salpn = the dianion of 1,3-bis-[(3,5-di-tert butylsalicylidene)amino]propane} to synthesize selectively complexes of different nuclearities. Four complexes have been synthesized: the mononuclear complex [Fe(3,5-(t)Bu(2)salpn)(HMeDCBI)] (1), the two binuclear but topologically different complexes [Fe(3,5-(t)Bu(2)salpn)(MeDCBI)Fe(3,5-(t)Bu(2)salpn)] (2) and {[Fe(3,5-(t)Bu(2)salpn)](2)(HDCBI)} (3), and the trinuclear complex {[Fe(3,5 (t)Bu(2)salpn)](2)(DCBI)Fe(3,5-(t)Bu(2)salpn)} (4). The structures of these complexes have been determined by X-ray crystallography. Variable-temperature direct-current magnetic susceptibility measurements were conducted for all compounds to obtain information about their electronic structure and to investigate the extent of magnetic communication among the Fe(III) centers. The results of these measurements allowed us to correlate the different structural motifs with the possible magnetic interactions that arise in multinuclear complexes of dicarboxyimidazoles. For 1, the room-temperature chi(M)T value reveals an S = (5)/(2) ground state. The data for the binuclear but topologically different complexes 2 and 3, and the trinuclear complex 4 suggest that weak intramolecular antiferromagnetic interactions are present, with interaction parameters ranging from -3.6 to -5.1 cm(-1). Differences in the extent of the magnetic communication between the metal centers through the two different interaction pathways of the ligands MeDCBI and DCBI (either through the imidazole ring or through the carboxylate groups) have been observed in complexes 2-4 that can be explained by the structural differences observed in the crystal structures of these compounds (the separation of the metal centers and the coplanarity of the metal ion orbitals with the pi system of the ligands). Cyclic voltammetry measurements for the mononuclear compound 1 show an irreversible reduction wave that is attributed to Fe(3+) + e(-) --> Fe(2+). The electrochemical behavior of the multinuclear complexes 2-4 is more complicated; however, it indicates that there is a degree of electronic communication between the Fe(III) centers. PMID- 15877448 TI - Square planar vs tetrahedral coordination in diamagnetic complexes of nickel(II) containing two bidentate pi-radical monoanions. AB - The reaction of three different 1-phenyl and 1,4-diphenyl substituted S methylisothiosemicarbazides, H(2)[L(1-6)], with Ni(OAc)(2).4H(2)O in ethanol in the presence of air yields six four-coordinate species [Ni(L(1-6)(*))(2)] (1-6) where (L(1-6)(*))(1-) represent the monoanionic pi-radical forms. The crystal structures of the nickel complexes with 1-phenyl derivatives as in 1 reveal a square planar structure trans-[Ni(L(1)(-3)(*))(2)], whereas the corresponding 1,4 diphenyl derivatives are distorted tetrahedral as is demonstrated by X-ray crystallography of [Ni(L(5)(*))(2)] (5) and [Ni(L(6)(*))(2)] (6). Both series of mononuclear complexes possess a diamagnetic ground state. The electronic structures of both series have been elucidated experimentally (electronic spectra magnetization data). The square planar complexes 1-3 consist of a diamagnetic central Ni(II) ion and two strongly antiferromagnetically coupled ligand pi radicals as has been deduced from correlated ab initio calculations; they are singlet diradicals. The tetrahedral complexes 4-6 consist of a paramagnetic high spin Ni(II) ion (S(Ni) = 1), which is strongly antiferromagnetically coupled to two ligand pi-radicals. This is clearly revealed by DFT and correlated ab initio calculations. Electrochemically, complexes 1-6 can be reduced to form stable, paramagnetic monoanions [1-6](-) (S = (1)/(2)). The anions [1-3](-) are square planar Ni(II) (d,(8) S(Ni) = 0) species where the excess electron is delocalized over both ligands (class III, ligand mixed valency). In contrast, one-electron reduction of 4, 5, and 6 yields paramagnetic tetrahedral monoanions (S = (1)/(2)). X-band EPR spectroscopy shows that there are two different isomers A and B of each monoanion present in solution. In these anions, the excess electron is localized on one ligand [Ni(II)(L(4-6)(*))(L(4-6))](-) where (L(4-6))(2-) is the closed shell dianion of the ligands H(2)[L(4-6)] as was deduced from their electronic spectra and broken symmetry DFT calculations. Oxidation of 1 and 5 with excess iodine yields octahedral complexes [Ni(II)(L(1,ox))(2)I(2)] (7), [Ni(II)(L(1,ox))(3)](I(3))(2) (8), and trans-[Ni(II)(L(5,ox))(2)(I(3))(2)] (9), which have been characterized by X-ray crystallography; (L(1-)(6,ox)) represent the neutral, two-electron oxidized forms of the corresponding dianions (L(1-6))(2 ). The room-temperature structures of complexes 1, 5, and 7 have been described previously in refs 1-5. PMID- 15877449 TI - Iminic N-bound iminophosphorano versus nitrilic n-bound iminophosphorano Os(IV) complexes: a new double derivatization of the nitrido ligand. AB - The reactions between trans-[Os(IV)(tpy)(Cl)(2)(NCN)] (1) and PPh(3) and between trans-[Os(IV)(tpy)(Cl)(2)(NPPh(3))](+) (2) and CN(-) provide new examples of double derivatization of the nitrido ligand in an Os(VI)-nitrido complex (Os(VI)N). The nitrilic N-bound product from the first reaction, trans [Os(II)(tpy)(Cl)(2)(NCNPPh(3))] (3), is the coordination isomer of the first iminic N-bound product from the second reaction, trans [Os(II)(tpy)(Cl)(2)(N(CN)(PPh(3)))] (4). In CH(3)CN at 45 degrees C, 4 undergoes isomerrization to 3 followed by solvolysis and release of (N cyano)iminophosphorane, NCNPPh(3). These reactions demonstrate new double derivatization reactions of the nitrido ligand in Os(VI)N with its implied synthetic utility. PMID- 15877450 TI - Acetic acid induced self-assembly of supramolecular compounds [Et4N]3[(WS4Cu2)2(mu-CN)3].2MeCN and [PPh4][WS4Cu3(mu-CN)2].MeCN from preformed clusters [A]2[WS4(CuCN)2] (A = Et4N, PPh4). AB - Reactions of two preformed trinuclear W/Cu/S clusters, [A](2)[WS(4)(CuCN)(2)] (1: A = Et(4)N; 2: A = PPh(4)), with different concentrations of acetic acid in MeCN generate two interesting 2D polymeric clusters [Et(4)N](3)[(WS(4)Cu(2))(2)(mu CN)(3)].2MeCN (3), and [PPh(4)][WS(4)Cu(3)(mu-CN)(2)].MeCN (4), respectively. Compound 4 can also be readily obtained in a high yield from the reaction of 2 with equimolar [Cu(MeCN)(4)]PF(6) in MeCN. These compounds have been characterized by elemental analysis, IR spectra, thermal analysis, and single crystal X-ray diffraction. An X-ray analysis reveals that compound 3 retains the WS(4)Cu(2) cluster core, which serves as a 3-connecting node to link equivalent nodes via single cyanide bridges, forming an anionic 2D (6,3) net. Compound 4 consists of a T-shaped WS(4)Cu(3) core, which also acts as a 3-connecting node, with links to 3 equivalent clusters either through single or double cyanide bridges, affording a different anionic 2D (6,3) network. The acetic acid induced aggregation of 3 and 4 from the two cluster precursors 1 and 2 suggests that this simple synthetic strategy is likely to be applicable to many related systems. PMID- 15877451 TI - Synthesis, structure, and characterization of new mononuclear Mn(II) complexes. Electrochemical conversion into new oxo-bridged Mn(2)(III,IV) complexes. Role of chloride ions. AB - Two Mn(II) complexes are isolated and X-ray characterized, namely, cis [(L(2))Mn(II)(Cl)(2)] (1) and [(L(3))Mn(II)Cl(OH(2))](ClO(4)) (2(ClO(4))), where L(2) and L(3) are the well-known tetradentate N,N'-dimethyl-N,N'-bis(2 pyridylmethyl)ethane-1,2-diamine and N,N'-dimethyl-N,N'-bis(2 pyridylmethyl)propane-1,3-diamine ligands, respectively. The crystal structure reveals that whereas the ligand L(2) is in the cis-alpha conformation in complex 1, the ligand L(3) is in the more unusual cis-beta conformation in 2. EPR spectra are recorded on frozen solutions for both complexes and are characteristic of Mn(II) species. Electrochemical behaviors are investigated on acetonitrile solution for both complexes and show that cation 2 exists as closely related Mn(II) species in equilibrium. For both complexes exhaustive bulk electrolyses of acetonitrile solution are performed at oxidative potential in various experimental conditions. In the presence of 2,6-lutidine and after elimination of chloride ligands, the formation of the di-mu-oxo mixed-valent complexes [(L(2))Mn(III)(mu-O)(2)Mn(IV)(L(2))](3+) (3a) and [(L(3))Mn(III)(mu O)(2)Mn(IV)(L(3))](3+) (4) is confirmed by UV-vis and EPR spectroscopies and cyclic voltammetry. In addition crystals of 4(ClO(4))(3) were isolated, and the X ray structure reveals the cis-alphaconformation of L(3). In the absence of 2,6 lutidine and without elimination of the exogenous chloride ions, the electrochemical oxidation of 1 leads to the formation of the mononuclear Mn(III) complex, namely, [(L(2))Mn(III)(Cl)(2)](+) (5), as confirmed by UV-vis as well as parallel mode EPR spectroscopy and cyclic voltammetry. In the same conditions, the electrochemical oxidation of complex 2 is more intricate, and a thorough analysis of EPR spectra establishes the formation of the binuclear mono-mu-oxo mixed-valent [(L(3))ClMn(III)(mu-O)Mn(IV)Cl(L(3))](3+) (6) complexes. Electrochemical conversion of Mn(II) complexes into mixed-valent Mn(2)(III,IV) oxo-bridged complexes in the presence of 2,6-lutidine is discussed. The role of the chloride ligands as well as that of L(3) in the building of oxo bridges is discussed. Differences in behavior between L(2) and L(3) are commented on. PMID- 15877452 TI - La9Sb16Br3 and Ce9Sb16Cl3: stars and stripes in rare earth halide and intermetallic compounds. AB - The title compounds were synthesized from Ln, LnX(3) (Ln = La, Ce; X = Cl, Br), and Sb under an Ar atmosphere at 950 degrees C. They crystallize in the space group P6(3)/m (No. 176) with lattice constants a = 21.232(5) and 20.862(2) Angstroms and c = 4.323(2) and 4.2728(7) Angstroms for La(9)Sb(16)Br(3) and Ce(9)Sb(16)Cl(3), respectively. The solids are the most metal-rich members in the reduced rare earth metal halide family and contain partial structures which are characteristic of reduced halides and intermetallic phases. These are the [Ln(6)X(6)](infinity) hexagon stars, Sb-centered [Ln(3)Sb](infinity) trigonal prismatic columns, and stripes of Sb square meshes. Computational analysis indicates that their electronic structure is valence-precise in the reduced halide part, but electron-deficient in the intermetallic part. Susceptibility and resistivity measurements reveal the metallic nature of the compounds. PMID- 15877453 TI - Rhenium(V) complexes with thiolato and dithiolato ligands: synthesis, structures, and monomerization reactions. AB - The new compound {(PhS)(2)ReO(mu-SPh)}(2), 1, was synthesized from Re(2)O(7) and PhSH and then used as the synthon for a number of hitherto unknown oxorhenium(V) compounds. Reactions between dithiols and 1 (2:1 ratio) afford {PhSReO(dt)}(2), where the dithiols, dtH(2), are 1,2-ethanedithiol (edtH(2)), 1,3-propanedithiol (pdtH(2)), 1,3-butanedithiol (pdtMeH(2)), 1,2-benzenedithiol (bdtH(2)), 2 (mercaptomethyl)thiophenol (mtpH(2)), and 2-mercaptoethyl sulfide (mesH(2)). Similar reactions carried out with a 3:1 ratio of dtH(2) to 1 afford [(ReO)(2)(dt)(3)], dt = edt, pdt. When NEt(3) was introduced prior to the 3:1 reaction between edtH(2) and 1, a compound containing an anionic complex was isolated, [PPh(4)][ReO(edt)(2)]. The new compounds were characterized analytically, spectroscopically, and crystallographically. The Re-O groups in two of the compounds, 1 and {ReO(mu-SPh)(bdt)}(2), exist in rare anti orientations; the others adopt the more familiar syn geometry, as discussed. Selected monomerization reactions of {PhSReO(dt)}(2) were also carried out: {PhSReO(dt)}(2) + 2L = 2[PhSReO(dt)L]. The rate for L = 4-phenylpyridine is given by v = {k(a)[L] + k(b)[L](2)} x [{PhSReO(dt)}(2)], as it is for the reactions of {MeReO(dt)}(2); for all of these compounds, the reaction proceeds nearly entirely by the third-order pathway. Values of k(b)/L(2) mol(-2) s(-1) at 25.0 degrees C are 5.8 x 10(2) (mtp), 2.97 x 10(3) (pdt), 4.62 x 10(5) (edt), and 3.87 x 10(5) (bdt). The rate law for the reactions of {PhSReO(dt)}(2) with L = PAr(3) is v = k(a)[L]/{1 + kappa[L]} x [{PhSReO(dt)}(2)]. For PPh(3), values at 25.0 degrees C of k(a)/L mol(-1) s(-1) (kappa/L mol(-1)) for {PhSReO(dt)}(2) are 9.64 x 10(-2) (1.87) for mtp, 3.43 x 10(-2) (0.492) for pdt, 1.91 (1.42) for edt, 1.84 x 10(-2) (0.82) for bdt, and 1.14 x 10(3) (10.6) for 1. Mechanisms are proposed that are consistent with the data obtained and with earlier work. PMID- 15877454 TI - High-valent imido complexes of manganese and chromium corroles. AB - The oxidation reaction of M(tpfc) [M = Mn or Cr and tpfc = tris(pentafluorophenyl)corrole] with aryl azides under photolytic or thermal conditions gives the first examples of mononuclear imido complexes of manganese(V) and chromium(V). These complexes have been characterized by NMR, mass spectrometry, UV-vis, EPR, elemental analysis, and cyclic voltammetry. Two X ray structures have been obtained for Mn(tpfc)(NMes) and Cr(tpfc)(NMes) [Mes = 2,4,6-(CH(3))(3)C(6)H(2)]. Short metal-imido bonds (1.610 and 1.635 Angstroms) as well as nearly linear M-N-C angles are consistent with triple M triple-bond NR bond formation. The kinetics of nitrene [NR] group transfer from manganese(V) corroles to various organic phosphines have been defined. Reduction of the manganese(V) corrolato complex affords phosphine imine and Mn(III) with reaction rates that are sensitive to steric and electronic elements of the phosphine substrate. An analogous manganese complex with a variant corrole ligand containing bromine atoms in the beta-pyrrole positions, Mn(Br(8)tpfc)(NAr), has been prepared and studied. Its reaction with PEt(3) is 250x faster than that of the parent tpfc complex, and its Mn(V/IV) couple is shifted by 370 mV to a more positive potential. The EPR spectra of chromium(V) imido corroles reveal a rich signal at ambient temperature consistent with Cr(V) triple-bond NR (d(1), S = 1/2) containing a localized spin density in the d(xy) orbital, and an anisotropic signal at liquid nitrogen temperature. Our results demonstrate the synthetic utility of organic aryl azides in the preparation of mononuclear metal imido complexes previously considered elusive, and suggest strong sigma-donation as the underlying factor in stabilizing high-valent metals by corrole ligands. PMID- 15877455 TI - Structural characterization of four members of the electron-transfer series [PdII(L)2)2]n (L = o-Iminophenolate derivative; n = 2-, 1-, 0, 1+, 2+). ligand mixed valency in the monocation and monoanion with S = (1)/(2) ground states. AB - The reaction of the ligand 2-(2-trifluoromethyl)anilino-4,6-di-tert-butylphenol, H(2)((1)L(IP)), and PdCl(2) (2:1) in the presence of air and excess NEt(3) in CH(2)Cl(2) produced blue-green crystals of diamagnetic [Pd(II)((1)L(ISQ))(2)] (1), where ((1)L(ISQ))(*)(-) represents the o-iminobenzosemiquinonate(1-) pi radical anion of the aromatic ((1)L(IP))(2-) dianion. The diamagnetic complex 1 was chemically oxidized with 1 equiv of Ag(BF(4)), affording red-brown crystals of paramagnetic (S = (1)/(2)) [Pd(II)((1)L(ISQ))((1)L(IBQ))](BF(4)) (2), and one electron reduction with cobaltocene yielded paramagnetic (S = (1)/(2)) green crystals of [Cp(2)Co][Pd(II)((1)L(ISQ))((1)L(IP))] (3); ((1)L(IBQ))(0) represents the neutral, diamagnetic quinone form. Complex 1 was oxidized with 2 equiv of [NO]BF(4), affording green crystals of diamagnetic [Pd(II)((1)L(IBQ))(2)](3)(BF(4))(4){(BF(4))(2)H}(2).4CH(2)Cl(2) (5). Oxidation of [Ni(II)((1)L(ISQ))(2)] (S = 0) in CH(2)Cl(2) solution with 2 equiv of Ag(ClO(4)) generated crystals of [Ni(II)((1)L(IBQ))(2)(ClO(4))(2)].2CH(2)Cl(2) (6) with an S = 1 ground state. Complexes 1-5 constitute a five-membered complete electron transfer series, [Pd((1)L)(2)](n) (n = 2-, 1-, 0, 1+, 2+), where only species 4, namely, diamagnetic [Pd(II)((1)L(IP))(2)](2-), has not been isolated; they are interrelated by four reversible one-electron-transfer waves in the cyclic voltammogram. Complexes 1, 2, 3, 5, and 6 have been characterized by X-ray crystallography at 100 K, which establishes that the redox processes are ligand centered. Species 2 and 3 exhibit ligand mixed valency: [Pd(II)((1)L(ISQ))((1)L(IBQ))](+) has localized ((1)L(IBQ))(0) and ((1)L(ISQ))(*)(-) ligands in the solid state, whereas in [Pd(II)((1)L(ISQ))((1)L(IP))](-) the excess electron is delocalized over both ligands in the solid-state structure of 3. Electronic and electron spin resonance spectra are reported, and the electronic structures of all members of this electron-transfer series are established. PMID- 15877456 TI - Self-assembly of chiral coordination polymers and macrocycles: a metal template effect on the polymer-macrocycle equilibrium. AB - The self-assembly of racemic and enantiopure binaphthyl-bis(amidopyridyl) ligands 1,1'-C(20)H(12){NHC(=O)-4-C(5)H(4)N}(2), 1, and 1,1'-C(20)H(12){NHC(=O)-3 C(5)H(4)N}(2), 2, with mercury(II) halides (HgX(2); X = Cl, Br, I) to form extended metal-containing arrays is described. It is shown that the self-assembly can lead to homochiral or heterochiral polymers or macrocycles, through self recognition or self-discrimination of the ligand units, and the primary materials can further self-assemble through hydrogen bonding between amide substituents. In addition, the formation of macrocycles or polymers can be influenced by the presence or absence of excess mercury(II) halide, through a template effect, and mercury(II) halide inclusion complexes may be formed. In one case, an unusual polymeric compound was obtained, with 1 guest HgX(2) molecule for every 12 mercury halide units in the polymer. PMID- 15877457 TI - A series of single, double, and triple Me2biim-bridged dinuclear, trinuclear, and polymeric complexes: syntheses, crystal structures, and luminescent properties. AB - Reaction of ZnCl(2) and Me(2)biim (Me(2)biim = N,N'-dimethyl-2,2-'-biimidazole) in acidic or neutral aqueous solutions gave the noncoordinated ZnCl(4).H(2)Me(2)biim (1) or the double Me(2)biim bridged [Zn(2)Cl(4)(mu Me(2)biim)(2)] (2). Use of CdX(2) (X = Cl, Br, I) instead of ZnCl(2) yielded the single Me(2)biim bridged one-dimensional coordination polymer [CdX(2)(mu Me(2)biim)](n) (X = Cl, 3; Br, 4; I, 5). The stacking of the infinite chains are dominated by C-H...X interactions in 3 and 4 but by I...I interactions in 5, responsible for their different crystal structures. Use of Zn(NO(3))(2) instead of ZnCl(2) produced the novel triple Me(2)biim-bridged [Zn(2)(mu Me(2)biim)(3)(H(2)O)(2)](NO(3))(4).H(2)O (6). The unprecedented hexa-Me(2)bim bridged trinuclear [Cd(3)(mu-Me(2)biim)(8)](2)(ClO(4))(12)(H(2)O)(6) (7) was obtained by using Cd(CH(3)CO(2))(2) in the presence of NaClO(4). Compounds 1-7 were characterized by X-ray crystallography and IR. Examination of photophysical properties of 1-7 indicates that the fluorescence emission of Me(2)biim has been effectively enhanced, quenched, or shifted in its metal complexes 1-7. PMID- 15877458 TI - Kinetic and thermodynamic characterization of the cobalt and manganese oxyhydroxide cores formed in horse spleen ferritin. AB - Horse spleen ferritin (HoSF) containing 800-1500 cobalt or 250-1200 manganese atoms as Co(O)OH and Mn(O)OH mineral cores within the HoSF interior (Co-HoSF and Mn-HoSF) was synthesized, and the chemical reactivity, kinetics of reduction, and the reduction potentials were measured. Microcoulometric and chemical reduction of HoSF containing the M(O)OH mineral core (M = Co or Mn) was rapid and quantitative with a reduction stoichiometry of 1.05 +/- 0.10 e/M forming a stable M(OH)(2) mineral core. At pH 9.0, ascorbic acid (AH(2)), a two-electron reductant, effectively reduced the mineral cores; however, the reaction was incomplete and rapidly reached equilibrium. The addition of excess AH(2) shifted the reaction to completion with a M(3+)/AH(2) stoichiometry of 1.9-2.1, consistent with a single electron per metal atom reduction. The rate of reaction between M(O)OH and excess AH(2) was measured by monitoring the decrease in mineral core absorbance with time. The reaction was first order in each reactant with second-order rate constants of 0.53 and 4.74 M(-1) min(-1), respectively, for Co- and Mn-HoSF at pH 9.0. From the variation of absorbance with increasing AH(2) concentration, equilibrium constants at pH 9.0 of 5.0 +/- 1.9 for Co-HoSF and 2.9 +/- 0.9 for Mn-HoSF were calculated for 2M(O)OH + AH(2) = 2M(OH)(2) + D, where AH(2) and D are ascorbic acid and dehydroascorbic acid, respectively. Consistent with these equilibrium constants, the standard potential for the reduction of Co(III)-HoSF is 42 mV more positive than that of the ascorbic acid reaction, while the standard potential of Mn(III)-HoSF is 27 mV positive relative to AH(2). Fe(2+) in solution with Co- and Mn-HoSF under anaerobic conditions was oxidized to form Fe(O)OH within the HoSF interior, resulting in partial displacement of the Co or Mn by iron. PMID- 15877459 TI - Structural increments for 11-vertex nido-phospha- and aza(carba)boranes and borates; dependence of energy penalties on the extent of Electron Localization. AB - Relevant structural features and corresponding energy penalties were determined that allow to easily estimate the relative stabilities of 11-vertex nido-phospha- and aza-substituted boranes, borates, carbaboranes, and carbaborates. For this purpose, density functional theory computations at the B3LYP/6 311+G(d,p)//B3LYP/6-31G(d)+ZPE level were carried out to determine the relative energies of 95 phospha- and 46 aza(carba)boranes and -borates. Energy penalties assigned to disfavoring structural features show additive behavior and excellent precision with respect to the computed results, as in the case of 6- and 11 vertex nido-carboranes and -borates. An unsubstituted phosphorus atom was found to possess energy penalties quite similar to those of the three-electron-donating H-C group. A bare nitrogen atom has energy penalties much larger than those of a bare phosphorus atom. Four-electron-donating RP and RN moieties, however, have even more adverse energy penalties. The disfavoring effects of heteroatoms in a borane cluster are determined by the amount of electron localization, that is, primarily by the number of skeletal electrons that formally originate from the heterogroup and secondarily by the electronegativity. Heteroatom energy penalties are independent of the type of the other heteroatoms present in the same cluster. Some novel phospha(carba)borane geometries with bare and exo-substituted phosphorus atoms in the same cluster have favorable thermodynamic stabilities competitive with those of known isomers. PMID- 15877460 TI - Synthesis, structure, and thermal properties of soluble hydrazinium germanium(IV) and tin(IV) selenide salts. AB - The crystal structures of two hydrazinium-based germanium(IV) and tin(IV) selenide salts are determined. (N(2)H(5))(4)Ge(2)Se(6) (1) [I4(1)cd, a = 12.708(1) Angstroms, c = 21.955(2) Angstroms, Z = 8] and (N(2)H(4))(3)(N(2)H(5))(4)Sn(2)Se(6) (2) [P, a = 6.6475(6) Angstroms, b = 9.5474(9) Angstroms, c = 9.8830(10) Angstroms, alpha = 94.110(2) degrees, beta = 99.429(2) degrees, gamma = 104.141(2) degrees, Z = 1] each consist of anionic dimers of edge-sharing metal selenide tetrahedra, M(2)Se(6)(4-) (M = Ge or Sn), separated by hydrazinium cations and, for 2, additional neutral hydrazine molecules. Substantial hydrogen bonding exists among the hydrazine/hydrazinium molecules as well as between the hydrazinium cations and the selenide anions. Whereas the previously reported tin(IV) sulfide system, (N(2)H(5))(4)Sn(2)S(6), decomposes cleanly to microcrystalline SnS(2) when heated to 200 degrees C in an inert atmosphere, higher temperatures (>300 degrees C) are required to dissociate selenium from 1 and 2 for the analogous preparations of single-phase metal selenides. The metal chalcogenide salts are highly soluble in hydrazine, as well as in a variety of amines and DMSO, highlighting the potential usefulness of these compounds as precursors for the solution deposition of the corresponding metal chalcogenide films. PMID- 15877461 TI - Low-dimensional ferromagnetic properties of SrCuV2O7 and BaCuV2O7. AB - The crystal structure of isostructural SrCuV(2)O(7) and BaCuV(2)O(7) consists of one-dimensional (1D) zigzag chains of Cu atoms with next-nearest-neighbor interaction. The main intrachain interaction was found to be ferromagnetic and estimated at 4.6 K (Hamiltonian H approximately -2J). SrCuV(2)O(7) and BaCuV(2)O(7) are new examples in the scanty family of 1D ferromagnets. Isothermal magnetization measurements at 0.08 K and specific heat data showed that MCuV(2)O(7) exhibits antiferromagnetic long-range ordering at T(N) = 1.36 K for SrCuV(2)O(7) and T(N) = 1.47 K for BaCuV(2)O(7). Spin-flop transitions were observed in the antiferromagnetic state at 0.08 K near 0.5 kOe in SrCuV(2)O(7) and 2 kOe in BaCuV(2)O(7). In air, SrCuV(2)O(7) and BaCuV(2)O(7) melted incongruently above 983 and 1018 K, respectively. PMID- 15877462 TI - Reproducibility across microwave instruments: preparation of a set of 24 compounds on a multiwell plate under temperature-controlled conditions. PMID- 15877463 TI - Combinatorial evaluation of the host-guest chemistry of star-shaped block copolymers. PMID- 15877464 TI - One-step construction of peptidomimetic 5-carbamoyl-4-sulfonyl-2-piperazinones. PMID- 15877465 TI - Quality control in combinatorial chemistry: determinations of amounts and comparison of the "purity" of LC-MS-purified samples by NMR, LC-UV and CLND. AB - The absolute purities of 20 purified samples from a combinatorial library have been determined by a new method that uses the DMSO sidebands [1J[13C-1H]] as an internal standard for quantification. The obtained absolute amounts are compared with the amounts of sample obtained by weighing, with the calculated weights obtained by chemiluminescent nitrogen detection (CLND) chromatography and with the relative purities obtained by LC-UV chromatography. PMID- 15877466 TI - A topologically segregated one-bead-one-compound combinatorial glycopeptide library for identification of lectin ligands. AB - A glycopeptide library containing more than 500,000 compounds has been constructed from a combination of Asn-linked carbohydrates using one-bead-one compound combinatorial methodologies. The library was encoded with peptide markers that were topologically segregated on the interior of the solid support to negate interference with carbohydrate/protein recognition during lectin screening. Both the peptide backbone and carbohydrate components were randomized, but the glycosamine was limited to position 3 at the center of the pentapeptide to evaluate the influence of the peptide backbone in lectin recognition. Of the four lectins that were evaluated, remarkable selectivity was observed with wheat germ agglutinin (WGA), which recognizes N-acetyl glucosamine (GlcNAc). From more than 80,000 possible combinations, only six ligands were identified, all possessing GlcNAc. These compounds were independently synthesized, characterized, and evaluated in solution. All six of the glycopeptides showed higher affinity for WGA than GlcNAc, with one having a 4-fold increase. Modeling studies indicate that the peptide backbone is capable of interacting with amino acids in the active site of WGA, but these interactions are not strongly correlated with activity, suggesting that the primary role of the peptide is to properly orient the sugar in the recognition process. PMID- 15877467 TI - A highly automated, polymer-assisted strategy for the preparation of 2 alkylthiobenzimidazoles and N,N'-dialkylbenzimidazolin-2-ones. AB - A multistep, polymer-assisted solution phase strategy for the highly automated (auto-PASP) synthesis of 2-alkylthiobenzimidazole and N,N'-dialkylbenzimidazolin 2-one libraries is presented. The approach incorporates in-line purification techniques to afford library products directly with high purities and is exemplified by the preparation of a 96-member 2-alkylthiobenzimidazoline library 1[1-12,1-8] and a 72-member N,N'-dialkylbenzimidazolin-2-one library 9[1-12,1-6]. PMID- 15877468 TI - Focused combinatorial library design based on structural diversity, druglikeness and binding affinity score. AB - The advent of focused library and virtual screening has reduced the disadvantage of combinatorial chemistry and changed it to a realizable and cost-effective tool in drug discovery. Usually, genetic algorithms (GAs) are used to quickly finding high-scoring molecules by sampling a small subset of the total combinatorial space. Therefore, scoring functions play essential roles in focused library design. Reported here is our initial attempt to establish a new approach for generating a target-focused library using the combination of the scores of structural diversity and binding affinity with our newly improved drug-likeness scoring functions. Meanwhile, a software package, named LD1.0, was developed on the basis of the new approach. One test on a cyclooxygenase (COX)2-focused library successfully reproduced the structures that have been experimentally studied as COX2-selective inhibitors. Another test is on a peroxisome proliferator-activated receptors gamma-focused library design, which not only reproduces the key fragments in the approved (thiazolidinedione) TZD drugs, but also generates some new structures that are more active than the approved drugs or published ligands. Both of the two tests took approximately 15% of the running time of the ordinary molecular docking method. Thus, our new approach is an effective, reliable, and practical way for building up a properly sized focused library with a high hit rate, novel structure, and good ADME/T profile. PMID- 15877469 TI - Optimization of MoVSb oxide catalyst for partial oxidation of isobutane by combinatorial approaches. AB - Optimization of the Mo-V-Sb mixed-oxide system for the selective oxidation of isobutane to methacrolein by true combinatorial methods primarily is intended to reduce the number of experiments in a broad parameter space. Therefore, an evolutionary approach based on a genetic algorithm has been chosen to screen three generations of 30 catalysts. With the help of automated sol-gel synthesis techniques, a high-throughput continuous flow reactor (16UPCFR), and appropriate software for experimental design, a new catalyst composition with improved performance has been obtained. Finally, the best catalysts were scaled-up to gram quantities and tested in a continuous-flow reactor unit that was equipped with four parallel reactors (4UPCFR). The final catalyst showed a significantly higher selectivity toward methacrolein at the same isobutane conversion, compared to the initial Mo8V2Sb90O(x) catalyst. PMID- 15877470 TI - Solid-phase syntheses of 6-arylpyridazin-3(2H)-ones. AB - The 3-chloropyridazine moiety was immobilized on a Wang resin, using two different methodologies. The first of these involved direct nucleophilic substitution of 3,6-dichloropyridazine with the alcoholate of Wang resin. The experimental conditions were optimized. The second method involved a Mitsunobu reaction between the Wang resin and 6-chloropyridazin-3-ol during which a problem of regioselectivity was observed. The so-obtained chloropyridazine-containing resins were subsequently reacted with various arylboronic acids under Suzuki conditions. Acid cleavage yielded 6-arylpyridazin-3(2H)-ones with high chemical purity. PMID- 15877471 TI - A versatile synthetic approach to peptidyl privileged structures using a "safety catch" linker. AB - Peptidyl privileged structures have been widely used by many groups to discover biologically active molecules. In this context, privileged substructures are used as "hydrophobic anchors", to which peptide functionality is appended to gain specificity. Utilization of this concept has led to the discovery of many different active compounds at a wide range of biological receptors. A synthetic approach to these compounds has been developed on a "safety-catch" linker that allows rapid preparation of large libraries of these molecules. Importantly, amide bond formation/cleavage through treatment with amines is the final step; it is a linker strategy that allows significant diversification to be easily incorporated, and it only requires the inclusion of an amide bond. In addition, chemistry has been developed that permits the urea moiety to be inserted at the N terminus of the peptide, allowing the same set of amines (either privileged substructures or amino acid analogues) to be used at both the N- and C-termini of the molecule. To show the robustness of this approach, a small library of peptidyl privileged structures were synthesized, illustrating that large combinatorial libraries can be synthesized using these technologies. PMID- 15877472 TI - 2-Methoxy-6-oxo-1,4,5,6-tetrahydropyridine-3-carbonitriles: versatile starting materials for the synthesis of libraries with diverse heterocyclic scaffolds. AB - Heterocyclic demonstration libraries for agrochemical screening were prepared from the common intermediates 2-methoxy-6-oxo-1,4,5,6-tetrahydropyridine-3 carbonitriles (1), using standard solution-phase techniques. A total of 18 screening libraries were prepared in good to excellent yields. Several members of these libraries were active in the first level of agrochemical screening, especially in the fungicide screen. PMID- 15877473 TI - The solid-phase Nicholas reaction: scope and limitations. AB - Two libraries of alpha-substituted alkynes has been prepared on solid phase using a sequential Sonogashira/Nicholas reaction approach. The scope of nucleophiles in the Nicholas reaction on solid phase has been investigated, including carbon, oxygen, nitrogen, sulfur, fluoride, and hydride nucleophiles. The conditions for the reaction sequence have been optimized in terms of Lewis acid, catalyst for the Sonogashira step, temperature, reaction time, and decomplexation method, enabling the five-step sequence to be performed in 1 day. PMID- 15877474 TI - Combinatorial solid-phase synthesis of 6-hydroxy-1,2,3,4-tetrahydro-beta carbolines from l-5-hydroxytryptophan. AB - A library of biologically relevant 6-hydroxy-tetrahydro-beta-carbolines (6-OH THBCs) based on the L-5-OH-tryptophan scaffold was prepared. A solid-phase synthesis was developed, utilizing aminomethyl polystyrene resin and solid-phase optimized reactions, such as Pictet-Spengler condensation. The library was designed such that three points of diversity would be readily introduced, making the strategy potentially suitable for generation of a large number of compounds. PMID- 15877475 TI - Trifluoroacetic anhydride-mediated solid-phase version of the Robinson-Gabriel synthesis of oxazoles. AB - A traceless solid-phase synthesis of oxazoles 4 via Robinson-Gabriel reaction of solid-supported alpha-acylamino ketones 2 has been achieved. The reaction requires that the cyclization precursor be linked to a benzhydrylic-type linker (compounds 2) and that trifluoroacetic anhydride be used as the cyclodehydrating agent. The solvent has a dramatic effect on the latter reaction, which goes to completion and follows a cyclative-type mechanism only when an ethereal solvent is used. Different synthetic routes have been investigated toward assembling compounds 2. The most straightforward one, which we have validated more extensively, comprises the reaction of Merrifield alpha-methoxyphenyl (MAMP) resin with an alpha-amino ketone to form compounds 1, which are, in turn, acylated. Other methodologies and strategies allowing for the synthesis of compounds 1 that have been investigated include direct alkylation of Rink amide resin; reductive amination of the latter with alpha-keto aldehydes; reaction of MAMP resin with alpha-amino alcohols, followed by oxidation; and protection of Rink amide resin with either 2,4-dinitrosulfonyl or allyl group, followed by alkylation and removal of protecting group. In addition, we disclose a novel variant of the Ugi four-component reaction that allows for the preparation of compounds 2 in a single synthetic step. PMID- 15877476 TI - Preparation of a fully substituted purine library. AB - A library of tetra-substituted purine analogues was readily prepared via parallel synthesis. This strategy relies on a key cyclization of a 4,5-diaminopyrimidine with either a carboxylic acid or its derivative to construct the 2,8,9 trisubstituted 6-chloropurine core. Further elaborations of this core allow the introduction of other diversity points. This methodology is demonstrated through the preparation of a 135-membered library of tetra-substituted purines in good yields and high purity. PMID- 15877477 TI - Formation of 4-aminopyrimidines via the trimerization of nitriles using focused microwave heating. AB - A series of substituted aliphatic nitriles have been trimerized to their corresponding pyrimidine structures under solvent-free conditions in the presence of catalytic quantities of potassium tert-butoxide using a focused microwave reactor. Multigram quantities of the corresponding 4-aminopyrimidines have been prepared in high yields and purity following a simple and scaleable protocol. PMID- 15877478 TI - The application of "click chemistry" for the decoration of 2(1H)-pyrazinone scaffold: generation of templates. AB - The "click chemistry" approach has been explored on the 2-(1H)-pyrazinone scaffold for the generation of pharmacologically interesting heterocyclic moieties. Huisgen 1,3-dipolar cycloaddition has been evaluated as the key step for the construction of the 1,2,3-triazole ring at the C-3 position of 2-(1H) pyrazinones. Two different pathways have been successfully evaluated: (1) via C-C or C-O linkage of the acetylenic part to the C-3 position of the 2-(1H) pyrazinone scaffold or (2) via azide introduction in the C-3 position. The subsequent application of "click chemistry" resulted in the formation of hitherto unknown skeletons. Microwave irradiation has successfully been applied in different steps of the sequence. PMID- 15877479 TI - The 3 "hypo's" of hospitalization. PMID- 15877480 TI - Irregular papillae pattern on tongue. Patient notes discomfort following spicy meals. PMID- 15877481 TI - Pain management in persons with dementia. BODIES mnemonic helps caregivers relay pain-related signs, symptoms to physicians and nursing staff. PMID- 15877482 TI - Chronic pain management in older adults: with coxibs under fire, what now? AB - Chronic non-cancer pain is notoriously undertreated, especially when the source cannot be identified by objective testing. Effective treatment often requires a combination of pharmacologic and non-pharmacologic approaches. This article describes current medication management of chronic pain, with particular attention to opioids. Acetaminophen and anti-inflammatories are first-line drugs for mild to moderate pain. For neuropathic pain, anticonvulsants are finding an increasing role, as are topical agents. Antidepressants are often advisable. Regarding opioids, the article addresses concerns about addiction potential; distinguishes between addiction and physical dependency; details the role of tolerance to different effects of opioids; and discusses their safety. With appropriate dosing, vigilant management, and careful tapering, opioids are a safe and effective choice for pain management in older adults. Appropriate follow-up guidelines are presented. PMID- 15877483 TI - Geriatric autoimmune diseases: systemic lupus erythematosus, Sjogren's syndrome, and myositis. AB - Systemic lupus erythematosus (SLE), Sjogren's syndrome (SS), and dermatomyositis (DM)/polymyositis (PM) may be encountered in geriatric patients due to improved survival rates in patients with younger ages of onset or from elderly-onset (EO) disease. EO disease accounts for up to 20% of patients affected by these disorders, and is typically insidious rather than acute. Whereas SS and DM/PM are considered autoimmune diseases with distinct organ specificity, SLE is a systemic disorder that may affect multiple organ systems. Commonly used clinical and laboratory criteria for defining and diagnosing these diseases were largely developed for patients age <65, and need to be modified in the geriatric patient. Therapeutic strategies include attention to ongoing drug regimens, medical comorbidities, and the roles of fatigue, depression, and arthropathy. Each disease may be responsive to low-dose corticosteroids, with a role for first or second-line immunosuppressives as steroid-sparing agents. PMID- 15877484 TI - Rural general practitioner preceptors--how can effective undergraduate teaching be supported or improved? AB - INTRODUCTION: General practitioners (GPs) as rural GP-Preceptors play an important role in medical student teaching within the Discipline of General Practice, University of Tasmania, Australia. As well a significant teaching role, they are responsible for apportioning 20% of the mark medical students receive in the General Practice examination at the end of the rotation. The contribution of the student-rural GP preceptor relationship to recruiting and retaining rural GPs has been acknowledged. Despite these important responsibilities, as honorary teaching staff rural GP preceptors receive no formal training in undergraduate education or assessment, nor did they receive formal peer support. METHOD: To address the lack of educational preparation of GP-preceptors, and in accordance with the stringent teaching and assessment standards of the Australian Medical School Accreditation, a tripartite approach was taken to educational skills development among rural GP-preceptors. The Discipline of General Practice conducted a needs-analysis of 64 rural GP-preceptors, and an evaluation by questionnaire of the teaching efficacy of those 64 rural GP-Preceptors and 76 urban GP-preceptors by 64 final-year medical students after their 3-week rural and urban GP attachments. The second intervention, based on the assessment and evaluation findings, was devised and implemented by the University Department of Rural Health. A series of workshops comprised the interdisciplinary educational and support program for GP preceptors, entitled Preceptor Onsite Preparation Program for Information Education and Support, or POPPIES. RESULTS: The rural GP preceptor needs analysis indicated that the majority of rural preceptors had no clear understanding of how what they taught fitted into the overall curriculum; they believed that their role as a clinical teacher had not been clearly defined by the program director; and also that undergraduate students had little understanding of what they needed to learn during their attachments. In contrast, preceptors believed they understood what students needed to learn; they were confident in performing the role of preceptor; and were familiar with adult learning principles, goal setting processes, effective student evaluation and the provision of appropriate performance feedback. Evaluation of feedback from students revealed that while rural GP preceptors performed well overall in regard to providing quality teaching and learning experiences, there was a significant spread of scores across all criteria, and approximately 15%-25% of students perceived various aspects of their attachment to be mediocre or poor. CONCLUSION: The combined results of the medical student questionnaire and rural GP-preceptor survey indicated both a need and a desire for educational skills development among rural GP-preceptors. On this basis, the interdisciplinary educational and support program for GP-preceptors (POPPIES) was developed and implemented as a series of workshops throughout rural Tasmania. Although no objective data are yet available about teaching outcomes as a result of POPPIES workshops, preliminary responses from attendee GP preceptors indicated that the workshops were effective in addressing educational needs, and in providing rural clinical teachers with professional teaching development. PMID- 15877485 TI - Outsourcing and benchmarking in a rural public hospital: does economic theory provide the complete answer? AB - INTRODUCTION: The ideology and pronouncements of the Australian Government in introducing 'competitive neutrality' to the public sector has improved efficiency and resource usage. In the health sector, the Human Services Department directed that non-clinical and clinical areas be market tested through benchmarking services against the private sector, with the possibility of outsourcing. These services included car parking, computing, laundry, engineering, cleaning, catering, medical imaging (radiology), pathology, pharmacy, allied health and general practice. Managers, when they choose between outsourcing, and internal servicing and production, would thus ideally base their decision on economic principles. Williamson's transaction cost theory studies the governance mechanisms that can be used to achieve economic efficiency and proposes that the optimal organisation structure is that which minimises transaction costs or the costs of exchange. Williamson proposes that four variables will affect such costs, namely: (i) frequency of exchange; (ii) asset specificity; (iii) environmental uncertainty; and (iv) threat of opportunism. This paper provides evidence from a rural public hospital and examines whether Williamson's transaction cost theory is applicable. METHOD: Case study research operates within the interpretivism paradigm and is used in this research to uncover why the outsourcing decision was made. Such research aims to study real-life experiences by examining the way people think and act and, in contrast to positivism, allows the interviewer to participate to better understand the details and features of the experiences. In the present research, individual interviews were conducted with managers of the hospital and owners and staff of the vendor organisations using semi- and unstructured questions to ascertain the extent of, and processes used in, outsourcing specific functional areas, and areas that were not outsourced. RESULTS: Pathology, radiology, dental technician services and lawn mowing were outsourced while food services was retained internally. The outsourcing of radiology was due to the hospital being unable (or unwilling) to finance new equipment and the problematical relationship between the existing radiologists, and hospital management and staff. Outsourcing resulted in increased staff morale, upgraded capital equipment and improved services. The outsourcing of pathology and dental technical services aimed to increase labour flexibility, thereby decreasing costs. Additional drivers in pathology were the changing nature of the funding arrangements rendering it profitable for the private sector to move into the provision of pathology and the increasing power of the medical scientists' union. The outsourcing of lawn mowing was simply to reduce costs. Food services was not outsourced because there was a lack of evidence that costs could be reduced. In addition, the existing relationships with food services staff were regarded as important because they had previously made immense changes to work practices, reduced staff numbers and decreased costs. CONCLUSION: Transaction costs are important when analysing how managers make the outsourcing decision, but the evidence from this case is that not all transaction costs are included in the decision, and that such costs are more complex than can be included in the type of analysis often undertaken by decision-makers. Taking into account Williamson's variables, the research shows that the outsourcing of services did not comply solely with the levels of transaction frequency or the requirement of asset specificity. In addition, opportunistic behaviour was evident on the part of all parties and was used in some cases as a reason for outsourcing, and in others to sway the decision to the manager's predisposed choice. A variety of arrangements were used to reduce environmental uncertainty, such as the transfer of staff to the contractor and the use of long-term contracts. Indeed the case shows that relationships between the hospital, its staff and the vendor are an important consideration that may not always be factored into an analysis that relies solely on transaction costs. PMID- 15877486 TI - Nutritional status comparison of rural with urban school children in Faisalabad District, Pakistan. AB - INTRODUCTION: Health and nutrition problems are the result of unsatisfactory food intake or severe and repeated infections, or a combination of both, and are closely linked to inadequate health services and an unhealthy environment. In Pakistan, there is a significant difference in the economic status, life style and nutrition between urban and rural populations, and between male and female children. Such differences affect the health and nutritional status of children. This study was undertaken in order to assess the nutritional and growth status of male and female school-going children aged 6-12 years from rural and urban areas in Pakistan. METHODS: Data were collected from male and female school-going children aged 6-12 years in and around Faisalabad, Pakistan. A total of 2042 children (853 male, 1189 female) were studied for being underweight, and for stunting and wasting. Of the males, 42.3% were from urban and 57.7% were from rural areas. Of the females, 72.7% were from urban and 27.3% were from rural areas. RESULTS: A total of 36.1% children were stunted and 45.3% were underweight; 25.2% were below standard weight for height. Among the urban children, 33% were below the standard for height (stunted), and 32.3% were below the standard for weight (underweight); 32.7% were wasted. Of the rural children, 40.9% were stunted, 64.7% were underweight and 33.3% were wasted. The rural female group was the most affected and malnourished with 61.8% stunted and 84% underweight; 67.1% were wasted. CONCLUSION: It is suggested that a comprehensive strategy be implemented in developing countries, especially Pakistan, in order to prevent infant and child under-nourishment. In addition to poverty alleviation, the underlying causes that work against food security, food safety, womens' education, safe water supply, sanitation and adequate health care must be addressed. PMID- 15877487 TI - Economic importance of the health-care sector in a rural economy. AB - INTRODUCTION: Leaders in rural communities around the world are struggling to provide primary health-care services. Common reasons for this situation include low population density, low incomes in that population, and inadequate medical payment policies. Per capita incomes in many rural Oklahoma communities, like rural areas in most other US States, are lower than the State average due to higher unemployment, higher proportion of income derived from transfer payments (government subsidies) and fewer higher paying employment opportunities. OBJECTIVE: The primary objective of this paper is to measure the economic impact that the health sector has on a community's economy. METHODS: A model, which measures the direct and secondary employment and income impacts of the health sector, is presented using Atoka, a rural county located in south-eastern Oklahoma, USA, as case study. The model is applied to the county's five health sector components (hospitals, doctors, nursing homes, pharmacies and others). Employment and payroll data are obtained for these sectors and the model measures the employment and payroll generated throughout the community as the health businesses and employees spend dollars in the community. RESULTS: The total employment impact of the health sector in Atoka County represents approximately 18.5% of the total non-farm employment. Secondary impacts of health-sector activity include the creation of jobs in other industries due to business and household spending. CONCLUSION: This study demonstrates the economic importance of the rural-health sector by quantifying its impacts on employment and payroll. The relationships among the existence of quality health care, industry attraction and quality of life for senior members of the rural community are suggested. By using a tool such as the IMPLAN model presented, rural community leaders are offered assistance to make important decisions about the provision of health-care services, particularly when the possible closure of the local hospital becomes an issue. PMID- 15877488 TI - Urban-rural differentials in maternal and child health in Andhra Pradesh, India. AB - INTRODUCTION: The unborn child is totally dependent on the mother; after birth, the child depends completely on the immediate social environment of the family and of the mother in particular. Recently the Indian Government changed its emphasis from family planning programs to family welfare programs. The intention was to promote the maternal and child health (MCH) programs to improve the health of mothers and young children. This study examined urban-rural differentials in MCH, and the factors influencing net change in MCH input, its utilisation and its output between the Indian National Family Health Survey (NFHS)-I and NFHS-II. METHODS: The NFHS data collected from 4276 ever-married women in the age group 13 49 years in 1992 (NFHS-I) and 4032 ever-married women in the age group 15-49 years in 1998 (NFHS-II) were analysed. RESULTS: The analysis revealed a positive net change in maternal and child health input, its utilisation and its output in the years 1992-1998. CONCLUSION: Andhra Pradesh was one of the most successful Indian States in providing MCH services, even though urban-rural differentials still exist. The Indian Government must take the necessary steps to improve MCH programs, including the provision of information and education campaigns, and sending dedicated health personnel to remote and inaccessible rural areas in order to reduce child mortality. PMID- 15877489 TI - Value of hospital-employed medical officers in rural Australia. AB - The author reports on a unique experience of the issues relating to recruitment and retention of specialist medical staff in a regional hospital in rural Tasmania, Australia. Based on a particular time of a high specialist medical staff resignation, the author identifies the importance to sustainability of rural medical specialists, of quality hospital-employed medical officer (HMO) support staff, a factor that has not been well recognized previously. It was found that HMOs, in particular senior or specialist-in-training HMOs, considerably reduced the workload of rural specialists, especially their after hours workload. A rating scale to assess the acceptability of after-hours workload for rural specialists, taking into account the impact of HMO support, was developed and is presented in the article. The presence of quality supporting HMOs was a vital recruitment and retention issue for rural medical specialists at that time and can be generalised to other rural situations and should be officially recognised. The observations made support increasing the number of senior or specialist-in-training-HMO positions in rural hospitals in order to retain and recruit more rural specialists. PMID- 15877490 TI - Health service utilisation among Filipino women in remote Queensland, Australia. AB - INTRODUCTION: This paper will discuss health issues among Filipinas (women born in the Philippines) living in remote and rural environments in Queensland in comparison to Filipinas living in other parts of the state. The sample was recruited as part of the University of Queensland component of the Australian Longitudinal Study of Women's Health. Access to health services will be discussed in terms of the framework outlined by McKinley et al: (i) Knowledge (Availability) is measured in terms of consumers' knowledge about the services and consumers' confidence in being able to use the medical care they require; (ii) Accessibility is considered in terms of their location to the consumers in relation to the services (distance) and being able to physically get to them (transport); (iii) Affordability is measured in terms of the extent to which cost is seen as a barrier to service use; (iv) Accommodation is measured in terms of the extent to which service delivery arrangements are able to accommodate consumers' needs and the extent to which doctors' attitudes are seen to be accommodating of those needs. It would be expected that cultural and linguistic issues would primarily affect Knowledge and Accommodation. Differences in Accessibility and Affordability would be expected to be affected by location. Access to services in rural areas may be more difficult and dependent on personal transport. METHODS: The sample, consisting of women born in the Philippines, was recruited though community organizations as part of the University of Queensland component of the Australian Longitudinal Study of Women's Health. Most of the women lived in urban or semi-urban areas (391) and 90 lived in areas designated as rural or remote areas by Queensland Health. The overall response rate was 87% and 83%. RESULTS: There were no differences between urban and rural and remote women in terms of knowledge. However women living in rural and remote areas had greater difficulty getting their information needs met. Consequently problems caused by structural limitations of health service delivery were often exacerbated by differences in expectations of health services and lack of understanding of methods to optimise access to care. CONCLUSION: Many of the barriers experienced by migrant women in rural/ remote areas mirror those of other Australian women. However the results suggest that better information about health services and self-management may improve the interface between service providers and their clients. PMID- 15877491 TI - Strongyloidiasis: an issue in Aboriginal communities. AB - Strongyloidiasis, a disease caused by the parasitic gut nematode (roundworm), Strongyloides stercoralis, has the highest prevalence in the world in rural and remote Aboriginal communities of northern Australia. With prevalences greater than 25%, these communities have rates of strongyloidiasis higher those in the worst affected developing countries where surveys have been recently conducted. Available data indisputably support that strongyloidiasis is more prevalent in rural and remote Aboriginal communities than in the mainstream Australian community. However control of strongyloidiasis has not been given a high priority by government health departments, with the result that Aboriginal people in remote and rural communities in Northern Australia are still suffering from a preventable and treatable disease. This article suggests that the only way to address the strongyloidiasis problem in Indigenous communities is to have strongyloidiasis recognised and addressed at the national level. One component of this must be making health departments responsible for establishing appropriate systems of effective treatment for and monitoring of patients with strongyloidiasis. PMID- 15877492 TI - Health status and participation in a range of activities in an Australian community. AB - INTRODUCTION: The aim of the research was to determine the relationship between levels of participation in a community and self-assessed health status of people in a rural and regional setting. METHOD: A cross-sectional design, using a mailed, self-administered questionnaire was used. Questionnaires were mailed to a random sample of people aged 18 years and over who were registered on the electoral roll of a regional city and rural area, the Barwon and Otway regions of Victoria, Australia. The sample consisted of 1752 participants: 990 females (57%), 739 males (42%) and 23 sex undisclosed (1%). The range of participants was 18-98 years, and the mean age was 50.53 years (SD = 17.19). RESULTS: Self assessed physical and mental health were measured using the SF-12 scale. Participants with low incomes, and those with low self-assessed physical and mental health scores, were significantly more likely than other participants to agree with one or more of the social isolation items, indicating that they experienced some social isolation. Low levels of participation in social, sports, leisure or support activities were associated with low self-assessed physical and mental health. Disengagement with the local community was associated with low levels of self-assessed mental health. While younger people were more likely than older people to participate in social, sports, leisure or support activities, they were less involved as members of their community. Females were more likely than males to have been involved in five or more sports, leisure or support activities. Participation in civic activities was associated with high income. Levels of participation in the four different types of activities were combined (social activities, sport, leisure or support activities, community and group activities, and civic activities). Participants classified as low participators were more likely to be older participants, to have a low income and to have low scores for both physical and mental health. CONCLUSION: An association was found between health and community participation in a range of activities, and between health and engagement with the community in this rural and regional population. These findings are consistent with those reported from similar research with a metropolitan population sample. The current research suggests that the groups of people of most concern in terms of low participation rates, are people who have low incomes, people aged over 65 years, people who may be defined as possessing poor physical health and people who may be defined as possessing poor mental health. The relationship between age, community participation and health is complex and needs further exploration because it is not known whether poor health reduces community participation or whether reduced community participation results in poor health. However, current research suggests that developing and implementing strategies to promote people's engagement with and involvement in their local community is one important way of promoting the health of the community as a whole. PMID- 15877493 TI - Albuminuria in a remote South Australian Aboriginal community: results of a community-based screening program for renal disease. AB - INTRODUCTION: The poverty, poor environmental living conditions and poor health standards experienced by Aboriginal Australians in some communities in rural and remote Australia have been described recently as 'fourth world'. For more than a century Aboriginal people have suffered the effects of dispossession of their land; destruction of their traditional culture and values; and exposure to infectious diseases, alcohol and the Western diet that is high in fat and sugar. Collectively these factors have contributed to the prevalence of chronic disease that afflicts Aboriginal people. In particular, renal disease has emerged during the last decade as a major contemporary health problem for Aboriginal Australians. According to the latest age- and sex-adjusted figures, Aboriginal people now have approximately nine-fold the risk of non-Aboriginal Australians of developing end-stage renal disease. In parts of Australia's Northern Territory, where Aboriginal people represent over 20% of the Territory's population, the rates of end-stage renal disease have been described as 'epidemic', reaching 2700 per million in the Tiwi Islands. In response to a request from the Umoona Tjutagku Health Service in mid 1997, the Renal Unit at Flinders Medical Centre, Adelaide, South Australia, formed a partnership with the health service to conduct a renal-disease screening program for adult members of the Umoona Community at Coober Pedy, a town 850 kilometres north of Adelaide. The partnership was later expanded to include screening for children (conducted by the Renal Unit at the Women's and Children's Hospital, Adelaide, South Australia). The community named the program 'The Umoona Kidney Project'. The Umoona community had recently experienced the dislocation of a number of its older people who suffered from advanced renal disease and were undergoing dialysis in a variety of centres in South Australia and the Northern Territory. As a result, the community had suffered social trauma. Consistent with the community's overall holistic approach to healthcare, the community wanted the renal program to provide a focus for community awareness of and knowledge about chronic disease, as well as to complement existing health programs. OBJECTIVES: The study objectives were to identify the prevalence of risk factors for renal disease, notably albuminuria, in adults from a remote Aboriginal community, and to examine the association of albuminuria with other risk factors; to empower Aboriginal health workers to self-manage a sustainable, community-controlled renal health program; and to assess the reliability and cultural acceptability of point-of-care technology for detecting renal disease. METHOD: The study was a three-year cross-sectional voluntary adult screening program (The Umoona Kidney Project). The study was performed as a partnership between the Flinders Medical Centre Renal Unit and the Umoona Tjutagku Health Service, and it involved nephrologists, medical scientists, Aboriginal health workers and clinical nurses. SETTING: Umoona Tjutagku Health Service, 850 km north of Adelaide. PARTICIPANTS: 158 adult members of the Umoona community: 58 males (37%; mean age = 43.8 years, range 23-78) and 100 females (63%; mean age = 39.6 years, range 18-72). MAIN OUTCOME MEASURES: First morning urine albumin : creatinine ratio measured by the Bayer DCA 2000 point-of-care analyser machine (Bayer Australia, Melbourne, Australia); lying and standing blood pressure; random blood glucose; body mass index; urinalysis. RESULTS: The study found that of screened adults, 29/149 (19%, 95% C.I. 13%-27%) had persistent microalbuminuria and 13/149 (9%, 95% C.I. 4% 14%) had persistent macroalbuminuria; 62/148 participants (42%, 95% C.I. 34%-50%) had overt hypertension; 35/145 participants (24%, 95% C.I. 17%-32%) had diabetes; 3 participants were newly diagnosed as having non-insulin dependent diabetes; 96/148 participants (65%, 95% C.I. 57%-73%) were either overweight or obese. Strong correlation was observed between the progression of albuminuria and age, all blood pressure categories, blood glucose, body mass index and an increasing number of risk factors. CONCLUSIONS: The Umoona Kidney Project identified a significant community burden of previously unknown incipient and established renal disease that required addressing via clinical- and community-based interventions. The DCA 2000 was a reliable instrument for detecting albuminuria on-site in the remote clinical location and was well accepted by Aboriginal health workers and community participants. PMID- 15877494 TI - Echidna: a Web-based community information database to assist students undertaking rural clinical placements. AB - The number of health-sciences students who spend time training in rural areas is increasing. Students undertaking rural placements have identified the need to be fully informed about the rural areas in which they are to be placed. To address this need, Echidna, a Web-based database containing rural-community information has been developed. The website brings fragmented information together in one easily navigable location for quick and accurate searching. Information provided in the website includes demographic data, health and community services information and relevant links. This paper examines the history of Echidna's development, data inclusion and data maintenance issues and discusses how students from medicine, nursing and other health disciplines, as well as other health professionals and organisations benefit from the resource. PMID- 15877495 TI - Evaluation of a rural community-based disability service in Queensland, Australia. AB - INTRODUCTION: The article outlines the evaluation framework devised for a semi formal disability service project in central Queensland, Australia, which was implemented using a community-based, participatory model. Utilising a service framework known as Community-Based Rehabilitation and an implementation strategy adapted from Participatory Rural Appraisal, this model is presented as a potential alternative for rehabilitation and disability services in the light of concerns that such services are poorly tailored and inadequate in remote and rural areas of Australia. METHOD: In recognition of the difficulty of evaluating such participatory and community-based initiatives, this evaluation was based on the analysis of large amounts of qualitative data from multiple sources, which were categorised against key themes drawn from the literature, using a SWOT analysis (strengths, weaknesses, opportunities and threats). It is suggested that this innovative and multifaceted evaluation methodology may have broader application. RESULTS AND CONCLUSIONS: Findings of the service evaluation indicated positive informal, community and social outcomes. Formal structural and organisational outcomes were found to be limited with a possibility of compromising the long-term viability of the initiative. Suggestions are made regarding the process of implementing similar research initiatives. The model may have application in similar rural community-based initiatives internationally. PMID- 15877496 TI - Rural health resource databases: a little nurturing goes a long way. AB - Access to rural health research information together with the type and availability of educational resources in rural areas, are important to rural health care providers, community members, researchers, students, planners and policy makers. The Rural Health Research Register (RHRR) focuses on current and recent research activity being undertaken in Australia in the field of rural health, while the Health Education Rural Remote Resources Database (HERRD) focuses on education courses and resources relevant to the practice and professional development of rural and remote health professionals throughout Australia. Early versions of these databases were established between 1992 and 1997, and in the period 1998-2001 both information resources were systematically updated through targeted promotion, registrations and the creation of web accessible search facilities. They continue to be maintained and updated. Detailed information is available by searching the RHRR and HERRD databases via the web or by contacting the relevant coordinator. This article examines some of the issues in developing and maintaining these resources and demonstrates the usefulness of their contents to rural healthcare workers. PMID- 15877497 TI - Compliance, culture, and the health of Indigenous people. AB - From the medical literature of the 70's to recent research, compliance has been named as an issue in the management of chronic disease, particularly in Indigenous contexts. Compliance can also be thought of as a universal problem that has different names in different contexts. In this paper compliance is described as a measure, with a numerator and a denominator, and progress towards better compliance is discussed in terms of manipulation of each element. The numerator is adherence, which has proven resistant to significant improvement, and the denominator is the advice and treatment, based on Western scientific medicine. Reduced to its most elemental terms, compliance compares the beliefs of patients with those of Western medicine. Proposed solutions to improve compliance have included moving to 'concordance' by reaching shared understanding of treatment goals, or forgetting compliance altogether and concentrating on institutional and organisational change to better align services with expectations of the population of interest. Indigenous Australian health care has been particularly beset with treatment failure as a result of 'poor compliance'. This paper suggests that compliance per se is not the problem. Rather, the extent to which it is apparent is a measure of dissonance between the belief systems. This gap between the culture of Western medicine and Indigenous culture should not be narrowed by the extinction of Indigenous belief, but by the development of a shared knowledge which includes Indigenous health beliefs and practices. PMID- 15877498 TI - Recruiting foreign doctors to South Africa: difficulties and dilemmas. AB - In 2003, the Medical and Dental Professional Board of the Health Professions Council of South Africa again made it possible for foreign-qualified doctors to obtain registration without sitting an examination. This eased the way for eligible non-South African doctors to work in South Africa in order to assist with the current staffing crisis in rural hospitals. This personal view examines the issues and difficulties related to foreign-trained doctors practising in South Africa, such as short-term practice vs long-term commitment; strategies to promote preparation of local doctors for rural hospital practice; the loss of recruited doctors to third-party countries; the principle of distributive justice in international medical training; and defining acceptable standards of training. The conclusions reached are that while doctors from developing countries, especially from neighbouring African nations, have historically brought a wealth of practical experience to South Africa's rural hospitals, there are also mutual benefits in recruiting from doctors trained in developed countries. The guiding principle is to obtain well-trained and experienced doctors who are able to function in a rural hospital until there are sufficient local-trained doctors. While the article focuses specifically on medical staffing in rural South Africa, the principles discussed apply equally to other health-care professionals and other, similar countries. PMID- 15877499 TI - Rural hospital focus: staffing. AB - Rural and Remote Health is committed to the task of providing a freely accessible, international, peer-reviewed evidence base for rural and remote health practice. Inherent in this aim is a recognition of the universal nature of rural health issues that transcends both regional interests and local culture. While RRH is already publishing peer-reviewed material, the Editorial Board believes many articles of potential worth are largely inaccessible due to their primary publication in small-circulation, paper-based journals whose readership is geographically limited. In order to augment our already comprehensive, international evidence base, the RRH Editorial Board has decided to republish, with permission, selected articles from such journals. This will also give worthwhile small-circulation articles the wide audience only a web-based journal can offer. The RRH editorial team encourages journal users to nominate similar, suitable articles from their own world region. First, then, in what RRH hopes will become a regular feature, is a series of articles from the prominent South African rural doctor, Professor Ian Couper. This article first appeared in South African Family Practice 2000; 22 (5), and is reproduced here in its original form, with kind permission of both publisher and author. This article introduced a regular column feature in SAFP, 'Rural hospital focus', and was entitled 'Staffing'. PMID- 15877500 TI - Rural hospital focus: no transport, no primary health care. AB - Rural and Remote Health is committed to the task of providing a freely accessible, international, peer-reviewed evidence base for rural and remote health practice. Inherent in this aim is a recognition of the universal nature of rural health issues that transcends local culture and regional interests. While RRH is already publishing such peer-reviewed material, the Editorial Board believes many articles of potential worth are largely inaccessible due to their primary publication in small-circulation, paper-based journals whose readership is geographically limited. In order to generate and make available a comprehensive, international evidence base, the RRH Editorial Board has decided to republish, with permission, selected articles from such journals. This will also give worthwhile small-circulation articles the wide audience only a web based journal can offer. The RRH RRH editorial team encourages journal users to nominate similar, suitable articles from their own world region. This article 'Rural hospital focus: No transport, no primary health care', is second in our series. It first appeared in South African Family Practice 2000; 22 (6), and is reproduced here in its original form, with kind permission of both publisher and author, prominent South African rural doctor, Professor Ian Couper. 'Rural hospital focus' was the title of the SAFP column which presented this article. PMID- 15877501 TI - Rural hospital focus: accommodation. AB - Rural and Remote Health is committed to the task of providing a freely accessible, international, peer-reviewed evidence base for rural and remote health practice. Inherent in this aim is a recognition of the universal nature of rural health issues that transcends both regional interests and local culture. While RRH is already publishing peer-reviewed material, the Editorial Board believes many articles of potential worth are largely inaccessible due to their primary publication in small-circulation, paper-based journals whose readership is geographically limited. In order to augment our already comprehensive, international evidence base, the RRH Editorial Board has decided to republish, with permission, selected articles from such journals. This will also give worthwhile small-circulation articles the wide audience only a web-based journal can offer. The RRH editorial team encourages journal users to nominate similar, suitable articles from their own world region. This article 'Rural hospital focus: accommodation', is third in our series. It first appeared in South African Family Practice 2000; 22 (7), and is reproduced here in its original form, with kind permission of both publisher and author, prominent South African rural doctor, Professor Ian Couper. 'Rural hospital focus' was the title of the SAFP column which presented this article. PMID- 15877502 TI - Rural origin and rural medical exposure: their impact on the rural and remote medical workforce in Australia. AB - Australia, like many countries, finds it difficult to recruit enough medical practitioners to live and work in rural and remote communities. Over the last decade the Australian Commonwealth Government has invested in a national strategy to train its medical workforce to encourage recruits to rural and remote general practice. This strategy is based on overseas experience that rural origin students, and those experiencing early and repeated rural exposure during training, are more likely to practise in a rural location. The importance of rural origin as a predictor of rural practice is well documented in the literature. More recent studies have tended to focus on rural exposure during both undergraduate and early postgraduate years, and on developing rural curricula in a multifaceted approach to medical training. All 11 medical schools in Australia have modified their selection criteria to encourage students from rural and remote locations, and have, to a varying degree, encouraged rural exposure in parallel with developing uniquely rural content in their curricula. Many of these initiatives are quite recent and have not yet been thoroughly evaluated against their success in addressing shortages in the rural and remote medical workforce. The aim of the review is to explore how the relationship between rural origin and rural exposure during undergraduate and postgraduate training and choice of practice location has underpinned initiatives in medical education in Australia in the years 1990-2003. PMID- 15877503 TI - Evidence-based rural workforce policy: an enduring challenge. PMID- 15877505 TI - Male sexual and reproductive health among the Aboriginal and Torres Strait Islander population. PMID- 15877504 TI - Development of a Rural Asthma Management Model, RAMM. AB - INTRODUCTION: The prevalence of asthma in Australia is increasing and places a significant cost burden on the community as well as reducing individuals' quality of life. In the late 1990s, asthma was the sixth National Health Priority in Australia and the prevalence of asthma in the Loddon Mallee region (LMR) of Victoria was approximately 1% higher than the State average. Four LMR local government areas had close to double the State average hospital admission ratios for asthma. AIM: The aim of this project was to develop a Regional Asthma Management Model (RAMM) and strategies for its implementation throughout the LMR, as a tool to implement a major health priority of both the Victorian State and Australian Commonwealth governments: to improve health outcomes for people with asthma. METHODS: A literature review was undertaken to identify best practice in asthma management for use as the basis of questions in workbooks designed to profile and compare current asthma management practice in the LMR. The workbooks were sent to all acute hospitals, community health centres and asthma educators in the LMR. The completed workbooks were returned and respondents elaborated on the workbook data at one of five subregional workshops. A survey was also undertaken to identify the range of asthma management strategies currently used by regional general practitioners (GPs) and to invite their views on ways to improve asthma management in the region. To gain consumer input into the RAMM a semi-structured group interview was held in an urban area and individual interviews were held in two rural areas in the region. A multidisciplinary reference group provided guidance to the project and a documentation design team was convened. RESULTS: Of the 19 workbooks sent to individual acute hospitals, 15 (78.9%) were completed and returned; 13 of 14 workbooks (92.8%) sent to individual community health centres were completed and returned. Fourteen of 15 asthma educators identified in the LMR were employed in the acute hospitals and community health centres that returned the workbooks; one asthma educator worked privately. Of the 215 GP surveys distributed, 38 surveys (17.6%) were returned. The majority of this small sample of GPs supported developing a uniform regional approach to asthma management based on NAC guidelines. Consumers interviewed suggested treating doctors, and/or EDs provide patients and carers with written instructions regarding acute asthma attacks and advice on management strategies for the ensuing 24-48 hours. A regional profile of asthma management practice was produced and compared with identified best practice. Gaps in practice and services were identified and responsive recommendations formulated. The National Asthma Campaign (NAC) guidelines were used as the basis for RAMM documentation, a package which consisted of a Regional Asthma Clinical Pathway and Emergency Department (ED) Package. CONCLUSION: The RAMM developed during the project provides documentation to assist best-practice asthma management by regional EDs and acute hospitals. The methodology and outcomes of the RAMM reflect the geography of the region, with multiple service providers from different locations managing a person with asthma across the primary, secondary and tertiary continuum. The RAMM methodology has the potential to be applied to other diseases and to other rural environments. Although the RAMM was designed for rural areas it could be easily adapted to suit the metropolitan environment. Implementation and evaluation of RAMM documentation is in progress. PMID- 15877506 TI - Facilitating recruitment of podiatrists to rural health in South Australia through a joint academic-clinical appointment. AB - Recruiting medical, nursing and paramedical or allied health staff to rural positions is an international problem. An example of this is the difficulty in recruitment and retention of podiatrists in rural South Australia. Increasing the proportion of undergraduate university students from rural areas is just one approach to the problem. It has also been established that students who undertake rural placements often return to a rural area to work on completion of their course. An innovative approach to addressing the recruitment and retention of rural podiatrists in South Australia involved the establishment of a joint rural academic-clinical position. The introduction of this joint position has enabled a mentored and supported increase in rural clinical exposure and education for undergraduate podiatry students. The aim of the appointment was to improve the recruitment of podiatrists to South Australian rural areas; however, an number of other benefits have become apparent. PMID- 15877507 TI - An exploration of the role that expert knowledge plays in the assessment of undergraduate clinical competence: registered nurses' experiences. AB - INTRODUCTION: This phenomenological study, conducted in rural Australia, explored the experiences of registered nurses (RNs) responsible for assessing the clinical competence of undergraduate nursing students. The purpose of the study was to gain insight into the experiences of a group of registered nurses who assess student competence by exploring how they perform the assessment process. A key assumption on which this study was based is that the participants are 'expert nurses', as defined by Benner. METHOD: Participants were recruited using purposive sampling from a population of registered nurses who assessed the clinical performance of undergraduate nursing students studying at a rural university in New South Wales, Australia. Individual unstructured interviews were conducted and audiotaped with the participants' permission. The analysed data were given to all participants to check for accuracy and validation and a thematic analysis of the data was conducted. RESULTS: Four themes were identified; the major theme, described in this article, was identified in all the narratives. The participants all acknowledged that they use their expert nursing knowledge to assist them when assessing the clinical competence of nursing students. The participants used a variety of terms to describe this type of knowledge such as intuition, instinct, gut feeling and 'just knowing'. CONCLUSION: While the findings of this study confirmed that experienced nurses unconsciously use their expert nursing knowledge when making decisions about students' competence, the findings also indicated a lack of awareness or underestimation of the value of expert clinical knowledge. These findings reinforce the need for further investigation to determine the role of expert nursing knowledge in the clinical competency assessment process. This is particularly significant for rural registered nurses employed in small health care facilities, who often assume the role of assessors of student clinical competence. PMID- 15877508 TI - Rural hospital focus: students. AB - Rural and Remote Health is committed to the task of providing a freely accessible, international, peer-reviewed evidence base for rural and remote health practice. Inherent in this aim is a recognition of the universal nature of rural health issues that transcends both regional interests and local culture. While RRH is already publishing peer-reviewed material, the Editorial Board believes many articles of potential worth are largely inaccessible due to their primary publication in small-circulation, paper-based journals whose readership is geographically limited. In order to augment our already comprehensive, international evidence base, the RRH Editorial Board has decided to republish, with permission, selected articles from such journals. This will also give worthwhile small-circulation articles the wide audience only a web-based journal can offer. The RRH editorial team encourages journal users to nominate similar, suitable articles from their own world region. PMID- 15877509 TI - Rural hospital focus: defining rural. AB - Rural and Remote Health is committed to the task of providing a freely accessible, international, peer-reviewed evidence base for rural and remote health practice. Inherent in this aim is a recognition of the universal nature of rural health issues that transcends both regional interests and local culture. While RRH is already publishing peer-reviewed material, the Editorial Board believes many articles of potential worth are largely inaccessible due to their primary publication in small-circulation, paper-based journals whose readership is geographically limited. In order to augment our already comprehensive, international evidence base, the RRH Editorial Board has decided to republish, with permission, selected articles from such journals. This will also give worthwhile small-circulation articles the wide audience only a web-based journal can offer. The RRH editorial team encourages journal users to nominate similar, suitable articles from their own world region. PMID- 15877510 TI - A community based, patient-centred, longitudinal medical curriculum. AB - The School of Rural Health is an initiative of the Australian Commonwealth Government, the University of New South Wales, Australia, and the Greater Murray Area Health Service. The school was established in February 2000 to facilitate the recruitment of doctors to and their retention in rural areas. The school is responsible for providing an education program for half of the three-year clinical component of the six-year undergraduate course. This article outlines the educational philosophies and methodologies employed in the development of a community-based, patient-centred, longitudinal approach to medical education. Although developed for and delivered in a rural setting, the curriculum could easily be adapted for implementation in an urban setting. The article presents a synopsis of experiences during the initial implementation of the curriculum, and it provides recommendations for future developments. PMID- 15877511 TI - Effect of repeated community-based ivermectin treatment on the intensity of onchocerciasis in Nigeria. AB - INTRODUCTION: Ivermectin (Mectizan) is an effective and well-tolerated microfilaricidal drug that has emerged as the drug of choice for large-scale treatment of onchocerciasis. Repeated doses of ivermectin is reported to lower the incidence of microfilardermia, while a single dose will markedly reduce skin microfilarial loads for up to 12 months with a transient fall in microfilarial level. AIM: The aim of the study was to evaluate the effect of repeated community based ivermectin treatment on the intensity of onchocerciasis in 10 onchocerciasis-endemic communities of Lade District, Pategi Local Government Area of Kware State, Nigeria. METHODS: The participants, a cohort of 330 men from whom parasitologic pre-treatment data were collected before mass ivermectin distribution commenced in 1992, were evaluated just before the fifth round of annual ivermectin treatment in August-September 1996, and 12 months after the fifth round in August-September 1997. Two bloodless skin snips were obtained from participants' left and right iliac crests and incubated for 24 h in an 0.5 mL microfuge tube containing 0.2 mL normal saline, before microscopic examination and enumeration of microfilariae. The community microfilarial load (CMFL), the most sensitive parasitologic indicator, was computed. RESULTS: Following four rounds of repeated annual treatments of onchocerciasis with ivermectin in the 10 communities studied, varying degrees of reduction in CMFL were observed in six of the 10 communities. These reductions were significant in three of the six communities at the first evaluation carried out just before the fifth round of annual ivermectin treatment (p = or <0.05). There was an increase in the remaining four communities, mainly attributed to poor annual acceptance and compliance with annual ivermectin treatment due to the occurrence of severe adverse reactions and absenteeism. Post-treatment evaluation (12 months after the previous evaluation), just before the sixth round of annual ivermectin treatment was preceded by community mobilization (enlightenment on the usefulness of ivermectin an assurance on its safety), especially in the four affected communities. This brought about reductions in the CMFL in the four communities that were not previously affected. Although the CMFL of these four communities was reduced, the CMFL of three of the communities did not fall below the 1992 pre treatment level but was below the level recorded 12 months earlier. One of these affected communities had its CMFL reduced below the 1992 pre-treatment CMFL. CONCLUSION: This study confirms the efficacy of ivermectin in the reduction of CMFL but exposes the danger of failing to monitor annual ivermectin treatment where community ivermectin-based control efforts are ongoing. The results obtained from this study are not as impressive as results reported from closely monitored scientific evaluations of ivermectin efficacy studies. PMID- 15877512 TI - Rural health focus: organising CPD for rural hospitals. AB - Rural and Remote Health is committed to the task of providing a freely accessible, international, peer-reviewed evidence base for rural and remote health practice. Inherent in this aim is a recognition of the universal nature of rural health issues that transcends both regional interests and local culture. While RRH is already publishing peer-reviewed material, the Editorial Board believes many articles of potential worth are largely inaccessible due to their primary publication in small-circulation, paper-based journals whose readership is geographically limited. In order to augment our already comprehensive, international evidence base, the RRH Editorial Board has decided to republish, with permission, selected articles from such journals. This will also give worthwhile small-circulation articles the wide audience only a web-based journal can offer. The RRH editorial team encourages journal users to nominate similar, suitable articles from their own world region. PMID- 15877513 TI - Overview of devolution of health services in the Philippines. AB - INTRODUCTION: In 1991 the Philippines Government introduced a major devolution of national government services, which included the first wave of health sector reform, through the introduction of the Local Government Code of 1991. The Code devolved basic services for agriculture extension, forest management, health services, barangay (township) roads and social welfare to Local Government Units. In 1992, the Philippines Government devolved the management and delivery of health services from the National Department of Health to locally elected provincial, city and municipal governments. AIM: The aim of this review is to (i) Provide a background to the introduction of devolution to the health system in the Philippines and to (ii) describe the impact of devolution on the structure and functioning of the health system in defined locations. METHOD: International literature was reviewed on the subjects of decentralization. Rapid appraisals of health management systems were conducted in both provinces. Additional data were accessed from the rural health information system and previous consultant reports. RESULTS: Subsequent to the introduction of devolution, quality and coverage of health services declined in some locations, particularly in rural and remote areas. It was found that in 1992-1997, system effects included a breakdown in management systems between levels of government, declining utilization particularly in the hospital sector, poor staff morale, a decline in maintenance of infrastructure and under financing of operational costs of services. CONCLUSION: The aim of decentralization is to widen decision-making space of middle level managers, enhance resource allocations from central to peripheral areas and to improve the efficiency and effectiveness of health services management. The findings of the historical review of devolution in the Philippines reveals some consistencies with the international literature, which describe some negative effects of decentralization, and provide a rationale for the Philippines in undertaking a second wave of reform in order to 'make devolution work'. PMID- 15877514 TI - Pilot study on the factors that influence learning by general practice registrars in central Australia. AB - INTRODUCTION: To counter a medical workforce shortage in rural and remote areas of Australia an increasing number of general practitioners are being trained in rural and remote areas. General practice (GP) registrars train in general practice as working apprentices alongside GP supervisors. GP registrars are allocated a training advisor to oversee their progress throughout their training. Central Australian GP registrars expressed concern to their training advisor regarding certain work partnerships with their GP supervisors. The study was carried out in response to these concerns, which were raised during a shortage of GPs in the area. The aim of the study was to explore factors in the interaction between GP registrars and GP supervisors in the context of their practices that impact on the quality of GP registrar learning in Central Australia. METHOD: A qualitative research method was used to explore the subtleties and issues in relationships between GP registrars, their GP supervisors and their practices. The interview schedule comprised pairs of polarised, provocative statements to generate discussion. Topics for the interview schedule were derived from the data from training advisor visits and the literature. GP registrars in Central Australia who had completed at least one six-month term in general practice were eligible for the study. Five female GP registrars participated in the study. Interviews were recorded, transcribed and checked by the participants before the interview material became the research record. RESULTS: The interview schedule generated considerable discussion as planned. The structures that determine GP income were seen as a barrier to GP registrar learning in Central Australia. The registrars reported that the fee-for-service model prevented them capitalising on learning opportunities both inside and outside their designated general practice. The GP registrars considered their training was compromised by the need to provide clinical service during a time of workforce shortage. Adaptation to a practice was seen as an important skill for GP registrars to learn, providing this did not compromise a registrar's own ethical and professional values. Learning was optimised by agreement between GP registrars and GP supervisors on the teaching subjects, and a mix of opportunistic and planned teaching sessions. Geographical isolation was perceived to have had a significant negative impact on GP registrar learning but one GP registrar discussed how this could be turned into a positive factor. CONCLUSION: GP registrars reported learning best by providing a clinical service with ready access to a supportive GP supervisor. Workforce pressures in Central Australia at the time of this pilot study reduced the GP supervisors' ability to support GP registrars, especially in a fee-for service model of health care. GP registrars should be placed in practices where they will receive experience, training and education rather than be allocated to areas of workforce shortage. Changes to the remuneration system for GP registrars and GP supervisors could be considered to enable GP registrars to capitalise on the learning opportunities in remote clinical practice. PMID- 15877516 TI - The ODAC Chronicles: Part 4. Hurdles pre and post accelerated approval. PMID- 15877518 TI - Docetaxel in the management of ovarian cancer. AB - Standard first-line treatment for Stage IC-IV ovarian cancer is currently a platinum agent or a combination of a platinum agent with a taxane. The use of a taxane compound in addition to single-agent platinum is increasingly preferred to platinum alone. In countries such as the UK, the taxane paclitaxel has been approved by the government for first-line use. However, it has yet to receive US Food and Drug Administration approval in the USA for use in this context. Typically, in countries such as the UK, patients with advanced ovarian cancer receive a combination of paclitaxel and carboplatin first line, both drugs given 3-weekly by intravenous infusion. Subsequent trials have demonstrated that the second-generation taxane docetaxel can be used as a substitute for paclitaxel; sharing many of its actions but with a different toxicity profile. However, docetaxel has not yet received approval for standard use. Here, the clinical development of docetaxel and its present and future place in the management of ovarian cancer is discussed. PMID- 15877519 TI - Paclitaxel for AIDS-associated Kaposi's sarcoma. AB - Treatment options are limited for patients with advanced acquired immunodeficiency syndrome-related Kaposi's sarcoma (AIDS-KS). The management of early stage cutaneous AIDS-KS has been revolutionized by the introduction of highly active antiretroviral therapy and for most patients highly active antiretroviral therapy alone will control early stage AIDS-KS. However, patients with advanced stage Kaposi's sarcoma with visceral disease, tumor-associated edema or extensive oral disease require systemic chemotherapy in addition to antiretrovirals. The standard first-line therapy for these affected individuals is a liposomal anthracycline, and response rates of around 70% are usually achieved. For patients with refractory or recurrent AIDS-KS, treatment algorithms are less well defined. The use of paclitaxel in these circumstances is reviewed. PMID- 15877520 TI - Clodronate in the prevention and treatment of skeletal metastasis. AB - Many solid tumors, including breast and prostate cancer, metastasize to bone, thereby putting patients at high risk for developing skeletal complications including pathologic fracture, spinal cord compression and debilitating bone pain. Patients often live for many years after developing bone metastasis, a fact that highlights the importance of therapies to reduce morbidity from skeletal complications. Bisphosphonates, including clodronate, have been shown to be useful in reducing skeletal complications in patients with cancer. This review will highlight the role of clodronate for skeletal metastasis. PMID- 15877521 TI - Pemetrexed-cisplatin combination in mesothelioma. AB - Despite the desperate struggle for improvement in the therapy of malignant pleural mesothelioma in previous decades, no standard therapy could be established. Pemetrexed (Alimta), a new multitargeted antifolate, showed significant antitumor activity in combination with cisplatin compared with cisplatin alone in patients with unresectable pleural mesothelioma. Furthermore, this improved efficacy was transformed into a significant symptomatic benefit. These data led to the approval of the drug by the US Food and Drug Administration and recently by the European Medicines Evaluation Agency for this combination in malignant pleural mesothelioma, and consequently founded the first standard of treatment in this disease. This review reflects the development of chemotherapy in malignant pleural mesothelioma, discusses the results of the EMPHACIS trial, and outlines further developments for pemetrexed-containing regimens in malignant pleural mesothelioma. PMID- 15877522 TI - MRI in the detection and management of breast cancer. AB - Breast magnetic resonance imaging (MRI) is now at a stage where the evidence is suggesting widespread potential in the management of patients with known or suspected breast cancers. MRI is used as a supplementary tool to complement conventional methods of breast evaluation because it has excellent problem solving capabilities. Many indications for clinical breast MRI are recognized, including resolving findings on mammography, staging of breast cancer when multiple or bilateral disease is suspected, and detecting the occult primary breast cancer presenting with malignant axillary lymphadenopathy but no detectable lesion on conventional breast examination. There is also encouraging ongoing research evaluating its role for the assessment of patients at high risk of breast cancer, for primary staging of cancers in radiographically dense breasts and for the assessment of response to chemotherapy. This article will review both the technical aspects of performing and interpreting breast MRI, as well as the current and possible future roles of breast MRI, comparing its strengths and weaknesses with conventional imaging. PMID- 15877523 TI - Brain metastases of breast cancer. AB - Brain metastases of breast cancer remain a difficult problem for clinical management. Their incidence appears to be increasing, which is likely due to longer survival times for advanced breast cancer patients as well as additional and improved tools for detection. Molecular features of tumors associated with this syndrome are not yet understood. In general, survival may be improving for brain metastases due to better local control in the CNS, as well as improvements in systemic disease management. Selected patients with brain metastases are able to undergo surgical resection, which has been associated with extended disease control in some patients. However, whole-brain radiation has been the mainstay for treatment for most patients. Stereotactic radiosurgery is playing an increasing role in the primary treatment of brain metastases, as well as for salvage after whole-brain radiation. Recent series have reported median survivals of 13 months or longer with stereotactic radiosurgery. Further improvements in radiation-based approaches may come from ongoing studies of radiosensitizing agents. The ability of systemic treatments to impact brain metastases has been debated, and specific treatment regimens have yet to be defined. New approaches include chemotherapy combinations, biologic therapies and novel drug-delivery strategies. PMID- 15877524 TI - Prognostic factors for breast cancer and their use in the clinical setting. AB - Breast cancer is the second leading cause of cancer deaths in US women today. This year, approximately 216,000 US women will be diagnosed with invasive breast cancer and another 60,000 with in situ disease. Numerous factors can quantify individual risks for breast cancer, guide therapy and predict outcome. This review focuses on the clinical, pathologic, molecular and genetic prognostic tools available for use in patients with breast cancer, and their impact on clinical decisions and treatment selection. PMID- 15877525 TI - Current chemotherapeutic strategies for rhabdomyosarcoma. AB - Rhabdomyosarcoma is a typical tumor of childhood and adolescence. Over the years there has been a gradual but important improvement in survival for patients with this tumor, despite its high grade of malignancy. These results are due to multidisciplinary treatment approaches including surgery, radiotherapy and especially chemotherapy. Rhabdomyosarcoma is a highly chemosensitive neoplasm, and the role of this therapeutic approach has also been clearly demonstrated in the adjuvant setting. This review covers current concepts on chemotherapy for rhabdomyosarcoma, with an overview of the results of the main clinical trials conducted over recent years and considerations of possible strategies for the near future. Recommendations for adult patients with rhabdomyosarcoma are also discussed, suggesting that these patients should be treated according to pediatric guidelines. PMID- 15877526 TI - Bisphosphonates and bone metastases: current status and future directions. AB - Since bone metastases in advanced cancer are common and frequently lead to skeletal-related morbid complications, their treatment remains a major challenge in cancer therapy. Bisphosphonates not only significantly decreased the odds ratios for fracture, need for radiotherapy, and incidence of hypercalcemia, but also had proven ability in the preservation of the 3D microstructure of bone that is responsible for bone stability. Bisphosphonates are well tolerated and have a very low incidence of serious side effects. Consequently, bisphosphonates have become the standard of care for the treatment of malignant bone disease. Benefits of bisphosphonate treatment appears to be more pronounced with longer treatment, indicating that they should be continued until no longer clinically relevant. As this advice has substantial implications on resources, it is essential that the use of bisphosphonates is evidence based. PMID- 15877527 TI - New concepts for the treatment of pediatric nonrhabdomyosarcoma soft tissue sarcomas. AB - Nonrhabdomyosarcoma soft tissue sarcomas form a group of rare tumors with a different biology and clinical behavior. The recently established European Pediatric Soft Tissue Sarcoma Study Group is organizing a new study devoted specifically to these tumors that were formerly treated according to the principles derived from experience with rhabdomyosarcoma, which is a clearly distinct entity. The new study includes two prospective trials, one for synovial sarcoma and the other for adult-type nonrhabdomyosarcoma soft tissue sarcomas. While surgery remains the mainstay of treatment, the role of adjuvant therapy is not yet clear and our understanding of the biology and natural history of nonrhabdomyosarcoma soft tissue sarcomas is still incomplete. This review presents the latest data on nonrhabdomyosarcoma soft tissue sarcoma treatment and outcome, and the rationale behind a risk-adapted treatment program that investigates the role of full-dose ifosfamide-doxorubicin chemotherapy in improving the response rate of patients with unresectable disease, the chances of avoiding adjuvant chemotherapy in low-risk synovial sarcomas, and the possible role of chemotherapy in high-risk adult-type soft tissue sarcomas. PMID- 15877528 TI - Current treatment for Ewing's sarcoma. AB - Ewing's sarcoma is the second most common primary bone tumor seen in children and adolescents, and was described by James Ewing in 1921 as a diffuse endothelioma of bone. It is one of the differential diagnoses of pediatric small round blue cell tumors. This is not a single condition, but a group of morphologically and clinically closely related disorders with similar molecular biology -- expression of tumor-specific chimeric oncoproteins through balanced chromosomal translocations involving the EWS gene -- often referred to as the Ewing family of tumors. This includes Ewing's sarcoma of bone, extra-osseous Ewing's sarcoma, Askin tumor and peripheral neuroectodermal tumor. These are aggressive neoplasms with almost 25% of patients having clinically evident metastases at presentation. Ewing's sarcoma has therefore been considered as a systemic disease necessitating local as well as systemic treatment. An aggressive multidisciplinary approach has resulted in significant improvement in prognosis for patients with these tumors. Despite aggressive treatment, 20-40% of patients with localized disease and almost 80% of patients with metastatic disease at presentation succumb to the illness. Advances in understanding the molecular biology of these tumors will hopefully result in the development of novel treatment approaches. The aim of this article is to review the existing treatment methods and to highlight the more recent approaches to the treatment of this condition. PMID- 15877529 TI - New opportunities in chemosensitization and radiosensitization: modulating the DNA-damage response. AB - Many current cancer treatments, including certain classes of chemotherapeutics and radiation, induce cytotoxicity by damaging DNA. However, many cancers are resistant to these therapies, which represents a significant challenge in the clinic. Thus, modulating DNA-damage responses to selectively enhance the sensitivity of cancer cells to these therapies is highly desirable. When DNA damage is detected, DNA checkpoint mechanisms are activated to halt cells at various phases of the cell cycle. Simultaneously, DNA-damage sensors transduce signals to activate DNA-repair mechanisms via de novo expression or post translational modification of enzymes required for DNA repair. p53 is the major player in a checkpoint that arrests cells at the G1/S boundary, while checkpoint kinase (Chk)1 is critical for the G2/M checkpoint and also the S checkpoint that prevents cell cycle progression after replication defects (intra-S-phase checkpoint) or S/M uncoupling (S/M checkpoint). Poly(ADP-ribose) polymerase is involved in sensing DNA single-strand breaks and inducing DNA repair via poly(ADP ribosyl)ating various DNA-binding and DNA-repair proteins. In this review, strategies for implementing small-molecule inhibitors of poly(ADP-ribose) polymerase and Chk1, which are emerging as potential adjuncts to current therapies, are discussed. PMID- 15877530 TI - Personalized medical treatment strategies for patients with chronic myeloid leukemia. AB - Management of patients with chronic myeloid leukemia has become increasingly difficult over the last few years since there are a variety of treatment options available. The ultimate challenge is to decide the most appropriate treatment strategy for an individual patient. To facilitate this, assessment of an individual patient's disease in terms of status, tempo and response to initial treatment needs to be determined. This review article discusses the current treatment options in the management of chronic myeloid leukemia, the factors that influence management decisions and suggests how treatment for the individual patient may be tailored whilst involving the patient in the decision-making process. PMID- 15877531 TI - Molecular neuro-oncology and the development of targeted therapeutic strategies for brain tumors. Part 5: apoptosis and cell cycle. AB - Brain tumors are a diverse group of malignancies that remain refractory to conventional treatment approaches. Molecular neuro-oncology has now begun to clarify the transformed phenotype of brain tumors and identify oncogenic pathways that might be amenable to targeted therapy. Abnormalities of the apoptotic and cell cycle signaling pathways are key molecular features of many brain tumors and are currently under evaluation for potential therapeutic intervention. The apoptosis pathway has numerous targets for molecular therapeutic development, including p53, Bax, Bcl-2, cFLIP, effector caspases, growth factor receptors, phosphatidylinositol-3-kinase, Akt and apoptosis inhibitors. Current molecular treatment approaches include antisense techniques, gene therapy and small molecule modulators and inhibitors. Potential targets of the cell cycle pathway include the cyclins, cyclin-dependent kinases, p53, retinoblastoma, E2F and the cyclin-dependent kinase inhibitors. Developmental molecular therapeutics for this pathway include adenoviral and gene therapy, small-peptide cyclin-dependent kinase modulators, proteasomal inhibitors and small-molecule cyclin-dependent kinase inhibitors. Several of these recently identified agents have begun evaluation in clinical trials. Further development of targeted therapies designed to modulate apoptosis and the cell cycle, and evaluation of these new agents in clinical trials, will be needed to improve survival and quality of life for patients with brain tumors. PMID- 15877532 TI - Multiple myeloma: an update of developments in targeted therapy. AB - Multiple myeloma continues to be an incurable illness. Improvement in our understanding of the disease is leading to new therapeutic modalities and the definition of new targeted therapy. Several pathways have been identified as areas for targeted therapy, and this review will discuss key molecules in some detail. New agents and molecules with biologic modulator activity appear to be changing the natural history of the disease. Agents such as thalidomide or its analogs, arsenic trioxide or bortezomib, hold great promise; however, much is yet to be learned on how to integrate these agents into disease management. PMID- 15877533 TI - Inverted papilloma involving the temporal bone and its association with squamous cell carcinoma: critical analysis of the literature. AB - Inverted papilloma is a rare, benign neoplasm, which usually originates from the lateral nasal wall and can be locally aggressive, extending into surrounding structures. Unusually, this disease involves temporal bone. There have been only 18 reported cases in the literature. The available data regarding inverted papilloma of the temporal bone are reviewed, and its etiopathogenesis, recurrence rate and association with squamous cell carcinoma are discussed. PMID- 15877535 TI - Yeast evolution and comparative genomics. AB - Recent sequencing efforts and experiments have advanced our understanding of genome evolution in yeasts, particularly the Saccharomyces yeasts. The ancestral genome of the Saccharomyces sensu stricto complex has been subject to both whole genome duplication, followed by massive sequence loss and divergence, and segmental duplication. In addition the subtelomeric regions are subject to further duplications and rearrangements via ectopic exchanges. Translocations and other gross chromosomal rearrangements that break down syntenic relationships occur; however, they do not appear to be a driving force of speciation. Analysis of single genomes has been fruitful for hypothesis generation such as the whole genome duplication, but comparative genomics between close and more distant species has proven to be a powerful tool in testing these hypotheses as well as elucidating evolutionary processes acting on the genome. Future work on population genomics and experimental evolution will keep yeast at the forefront of studies in genome evolution. PMID- 15877536 TI - Educating the staff at a homeless shelter about mental illness and anger management. AB - The purpose of this project was to assess the educational needs of the staff working at an urban homeless shelter for women and children, in terms of their knowledge about mental illness, and to provide educational sessions at the site based on the needs assessment. Needs were determined through a variety of sources including a survey designed to assess knowledge about mental illness and perceived learning needs, observations of staff interactions, and discussions with the director of the shelter. The ultimate goal was to have a positive impact on the treatment outcomes of the residents at this shelter by increasing the knowledge and skill level of the staff regarding working with clients with mental health conditions and with clients who exhibit angry behaviors. Most staff members interviewed several weeks after the educational sessions demonstrated application and retention of the information taught. PMID- 15877537 TI - Participation in community health screenings: a qualitative evaluation. AB - Community health screenings and education programs have been widely used to identify individual health risks and promote early diagnosis and treatment of disease. Participation in these community activities is dependent on the initial health-seeking behaviors of the individual. Focus group interviews1 were conducted with both attendees and nonattendees of a program offering health education and screenings in various settings in a semirural area. The purpose of these group interviews was to better understand issues associated with participation. A domain analysis of the qualitative data revealed that multiple aspects of self-care orientations as well as interpersonal and environmental factors influenced participation. A paradoxical facilitator-inhibitor effect was apparent. The companionship and support inherent in a community setting was an incentive for participation, but also presented concerns about confidentiality. However, concern about getting knowledge about self-care and the fear and anxiety of learning the results of screening tests creates a push-pull dilemma that is a challenge for health care providers. Factors that influence participation in semirural community health screenings are varied and complex and suggest multidimensional approaches be used in designing programs. PMID- 15877538 TI - Effects of a nutrition education program for urban, low-income, older adults: a collaborative program among nurses and nursing students. AB - Nutrition education programs can be effective in reducing risks of cardiovascular disease in older adults. The objective of the Eat and Learn Nutrition Program was to increase knowledge of nutrition and promote healthy eating among the residents of an older adult, low-income, urban housing community. The program was a series of 3 discussions on nutrition topics, based on a community needs assessment, that were presented in the common room of the participants' residence over a heart healthy lunch. On average, participants were able to increase their nutrition knowledge learning 1 major point per session. In addition, participants discussed alternative methods for healthy eating and shared culturally diverse nutritious recipes with each other. The collaborative nursing effort in designing the program served to facilitate learning about community program implementation for all nursing students involved. Collaboration between nurses and nursing students from a variety of educational backgrounds may enhance learning and program success. PMID- 15877539 TI - Orphan care in Malawi: current practices. AB - HIV and AIDS has moved rapidly throughout sub-Saharan Africa, dramatically adding to the crisis of orphaned children on the continent. Knowledge of African responses to their problem is needed so that interventions from the global community are culturally appropriate. An assessment of 73 programs caring for over 100,000 vulnerable and orphaned children in Malawi was conducted. A cross section of programs throughout the country was visited. Three primary care strategies were found: community-based orphan care, institutional and residential care, and self-care. The model of care preferred by Africans is community based because this keeps a child in a family environment in their own village and tribe. By listening to the people of Africa, the worldwide community can learn how to work with them as they care for millions of orphaned children. PMID- 15877540 TI - First trimester predictors of diet and birth outcomes in low-income pregnant women. AB - The purpose1 of this study is to test a model describing the relations of various biopsychosocial, behavioral, and cognitive factors on the health outcomes of nutritional adequacy and infant birth weight in low-income pregnant women. Descriptive, cohort design assessed low-income women (N=55) in their 1st trimester of pregnancy. Eighty percent of the women did not meet the recommended number of food servings outlined in the Food Guide Pyramid. Prepregnant body mass index (BMI) and pregnancy-related dietary changes explained 19% of the variance in nutritional adequacy. Maternal age, prepregnant BMI, nutritional knowledge, and infant gestation at birth explained 52% of the variance in infant birth weight. Nutritional adequacy in the 1st trimester and infant birth weight were not significantly related. Most low-income women are not meeting the nutritional requirements of pregnancy. Careful monitoring of dietary quality may help target women in need of additional nutritional education. Providing nutritional education throughout pregnancy may lead to improved dietary patterns that may reduce low birth weight. PMID- 15877544 TI - Hydrolysis characteristics of a beta-1,3-D-glucan elicitor from yeast. AB - Fluorophore-assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharides in a(n) (oligo)saccharide mixture. The single terminal aldehydes of oligoglucoside residues released by acid hydrolysis of beta 1,3-D-glucan from yeast were tagged with the charged fluorophore ANTS (8 aminonaphthalene-1,3,6-trisulphonate), and separated with high resolution on the basis of size by PAGE. ANTS fluorescence labelling was not biased by oligoglucoside length; therefore band fluorescence intensity was directly related to the relative abundance of individual oligoglucoside moieties in a heterogeneous sample. FACE analysis revealed that the major oligoglucoside mixture released by acid hydrolysis from beta-1,3-D-glucan was composed of monosaccharide, disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, hexasaccharide, heptasaccharide and octasaccharide, and the order of abundance from high to low was trisaccharide, monosaccharide, disaccharide, tetrasaccharide, pentasaccharide, hexasaccharide, heptasaccharide and octasaccharide respectively. In conclusion, FACE represents an accessible, sensitive and quantitative analytical tool enabling the characterization of a(n) (oligo)saccharide mixture. PMID- 15877545 TI - Dietary iron deficiency induces ventricular dilation, mitochondrial ultrastructural aberrations and cytochrome c release: involvement of nitric oxide synthase and protein tyrosine nitration. AB - Iron deficiency is associated with multiple health problems, including the cardiovascular system. However, the mechanism of action of iron-deficiency induced cardiovascular damage is unclear. The aim of the present study was to examine the effect of dietary iron deficiency on cardiac ultrastructure, mitochondrial cytochrome c release, NOS (nitric oxide synthase) and several stress-related protein molecules, including protein nitrotyrosine, the p47phox subunit of NADPH oxidase, caveolin-1 and RhoA. Male weanling rats were fed with either control or iron-deficient diets for 12 weeks. Cardiac ultrastructure was examined by transmission electron microscopy. Western blot analysis was used to evaluate cytochrome c, endothelial and inducible NOS, NADPH oxidase, caveolin-1 and RhoA. Protein nitrotyrosine formation was measured by ELISA. Rats fed an iron deficient diet exhibited increased heart weight and size compared with the control group. Heart width, length and ventricular free wall thickness were similar between the two groups. However, the left ventricular dimension and chamber volume were significantly enhanced in the iron-deficient group compared with controls. Ultrastructural examination revealed mitochondrial swelling and abnormal sarcomere structure in iron-deficient ventricular tissues. Cytochrome c release was significantly enhanced in iron-deficient rats. Protein expression of eNOS (endothelial NOS) and iNOS (inducible NOS), and protein nitrotyrosine formation were significantly elevated in cardiac tissue or mitochondrial extraction from the iron-deficient group. Significantly up-regulated NADPH oxidase, caveolin-1 and RhoA expression were also detected in ventricular tissue of the iron-deficient group. Taken together, these results suggest that dietary iron deficiency may have induced cardiac hypertrophy characterized by aberrant mitochondrial and irregular sarcomere organization, which was accompanied by increased reactive nitrogen species and RhoA expression. PMID- 15877546 TI - Spotting new DNA damage-responsive chromatin-binding proteins. AB - In response to DNA damage, cells initiate multiple repair mechanisms that all contribute to the survival of both the cell and the organism. These responses are numerous and variable, and can include cell cycle arrest, transcriptional activation of DNA repair genes and relocalization of repair proteins to sites of DNA damage. If all else fails, in multicellular organisms the initiation of apoptosis is also a potential cellular response to DNA damage. Despite a wealth of information about these events, it is clear that we do not yet have a comprehensive picture of the cellular responses to DNA damage. In this issue of the Biochemical Journal, a proteomics approach was used by Lee et al. to identify proteins that bind to chromatin in a DNA damage-inducible manner. The proteins identified, nucleophosmin, hnRNP C1 (heterogeneous nuclear ribonucleoprotein C1) and hnRNP C2, were proteins that would not necessarily have been predicted to behave this way. These studies have the potential to be extended and contribute to our knowledge of the cellular response to DNA damage. PMID- 15877547 TI - Self-assembly of SNARE fusion proteins into star-shaped oligomers. AB - Three evolutionarily conserved proteins known as SNAREs (soluble N-ethylmaleimide sensitive fusion protein attachment protein receptors) mediate exocytosis from single cell eukaryotes to neurons. Among neuronal SNAREs, syntaxin and SNAP-25 (synaptosome-associated protein of 25 kDa) reside on the plasma membrane, whereas synaptobrevin resides on synaptic vesicles prior to fusion. The SNARE motifs of the three proteins form a helical bundle which probably drives membrane fusion. Since studies in vivo suggested an importance for multiple SNARE complexes in the fusion process, and models appeared in the literature with large numbers of SNARE bundles executing the fusion process, we analysed the quaternary structure of the full-length native SNARE complexes in detail. By employing a preparative immunoaffinity procedure we isolated all of the SNARE complexes from brain, and have shown by size-exclusion chromatography and negative stain electron microscopy that they exist as approx. 30 nm particles containing, most frequently, 3 or 4 bundles emanating from their centre. Using highly purified, individual, full-length SNAREs we demonstrated that the oligomerization of SNAREs into star-shaped particles with 3 to 4 bundles is an intrinsic property of these proteins and is not dependent on other proteins, as previously hypothesized. The average number of the SNARE bundles in the isolated fusion particles corresponds well with the co-operativity observed in calcium-triggered neuronal exocytosis. PMID- 15877548 TI - Chronic cytomegalovirus infection and inflammation are associated with prevalent frailty in community-dwelling older women. AB - OBJECTIVES: To evaluate the association between asymptomatic chronic cytomegalovirus (CMV) infection and the frailty syndrome and to assess whether inflammation modifies this association. DESIGN: Cross-sectional analysis. SETTING: Women's Health and Aging Study I & II, Baltimore, Maryland. PARTICIPANTS: Seven hundred twenty-four community-dwelling women aged 70 to 79 with baseline measures of CMV, interleukin-6 (IL-6), and frailty status. MEASUREMENTS: CMV serology and IL-6 concentrations were measured using enzyme linked immunosorbent assay. Frailty status was based on previously validated criteria: unintentional weight loss, weak grip strength, exhaustion, slow walking speed, and low level of activity. Frail women had three or more of the five components, prefrail women had one or two components, and women who were not frail had none of the components. Multinomial logistic regression adjusted for potential confounders. RESULTS: Eighty-seven percent of women were CMV seropositive, an indication of chronic infection. CMV was associated with prevalent frailty, adjusting for age, smoking history, elevated body mass index, diabetes mellitus, and congestive heart failure (CMV frail adjusted odds ratio (AOR)=3.2, P=.03; CMV prefrail AOR=1.5, P=.18). IL-6 interacted with CMV, significantly increasing the magnitude of this association (CMV positive and low IL-6 frail AOR=1.5, P=.53; CMV positive and high IL-6 frail AOR=20.3, P=.007; CMV positive and low IL-6 prefrail AOR=0.9, P=.73; CMV positive and high IL-6 prefrail AOR=5.5, P=.001). CONCLUSION: Chronic CMV infection is associated with prevalent frailty, a state with increased morbidity and mortality in older adults; inflammation enhances this effect. Further prospective studies are needed to establish a causal relationship between CMV, inflammation, and frailty. PMID- 15877549 TI - Pneumonia versus aspiration pneumonitis in nursing home residents: prospective application of a clinical algorithm. AB - OBJECTIVES: To prospectively evaluate a clinical algorithm for the diagnosis of pneumonitis and pneumonia in nursing home residents. DESIGN: Prospective cohort study. SETTING: Inpatient geriatrics unit. PARTICIPANTS: Nursing home residents admitted to the hospital with suspected pneumonia. MEASUREMENTS: Identification of pneumonitis and pneumonia using the algorithm; medical record review and abstraction of clinical data; hospital outcome and length of stay. RESULTS: One hundred seventy episodes of suspected pneumonia were screened with the algorithm and classified into four groups: 25% pneumonia, 28% aspiration pneumonitis of 24 hours or less duration, 12% aspiration pneumonitis of more than 24 hours' duration, and 35% an aspiration event without pneumonitis. Presenting symptoms and signs, laboratory tests, severity of illness measures, or serum C-reactive protein levels did not distinguish between the four groups. Those with an aspiration event without pneumonitis tended to be treated less often with antibiotic therapy after admission (P=.004) and after discharge (P=.01). Of those who survived, there was no significant difference in mean hospital length of stay between the four groups. There was no significant difference in the percentage of case fatality between the four groups, but those with aspiration pneumonitis of 24 hours or less duration and with an aspiration event without pneumonitis had a lower mortality than the other two groups. CONCLUSION: Distribution of episodes of suspected pneumonia by clinical category as determined using the algorithm was similar to that of the derivation study, as were case fatality rates in each category. These findings suggest that the algorithm may be useful for making the distinction between pneumonitis and pneumonia in nursing home residents; further studies are warranted. PMID- 15877551 TI - The effect of hospitalization with medical illnesses on the suicide risk in the oldest old: a population-based register study. AB - OBJECTIVES: To compare the effect of hospitalization with medical illnesses on the suicide risk in the oldest old (> or = 80) with that in the old (65-79) and middle-aged (52-64) using nationwide data. DESIGN: Event-history analysis using time-varying covariates based on prospective individual-level register data. SETTING: Population-based record linkage. PARTICIPANTS: All persons aged 52 and older living in Denmark during 1996 to 1998 (N=1,684,205). MEASUREMENTS: The studied event is completed suicide. The following time-varying variables are included in the analysis: current age, hospitalization with medical illnesses within previous 2 years, and number of different medical diagnoses given at hospitalizations within previous 2 years. Relative suicide risks were calculated using event-history analysis. The proportional attributable risk was calculated to assess to which extent hospitalization can explain the increased suicide risk in the oldest old. RESULTS: During the 3-year study period, 1,184 persons committed suicide. Hospitalization with medical illnesses was associated with an increased suicide risk in the study population. The highest suicide rates were found in the oldest-old men who had been hospitalized during the previous 2 years: 113 per 100,000 versus 80 per 100,000 in the general population of men aged 80 and older. Also, the oldest-old women with a history of hospitalization had a higher suicide rate than their peer group. Nevertheless, the oldest old experienced a lower increase in risk after hospitalization than the middle-aged. Experiencing three or more different diagnoses increased the risk further. Almost two-thirds of the oldest old who committed suicide had experienced a medical hospitalization during a 2-year period preceding the suicide. A substantial part of the greater suicide rate in the oldest old than in the middle-aged can be attributed to the increased prevalence of medical hospitalization. CONCLUSION: The oldest old who have been hospitalized with medical illnesses have a significantly higher suicide risk than people in the same age range with no hospitalization, although they experience a lower increase in risk after hospitalization than the middle-aged. The increased prevalence of medical illnesses explains a part of the greater suicide risk with age, especially for the oldest old women. Considering that hospitalization with medical illness often precedes suicide in the oldest old, hospitalization may play an important role in identification of suicidal ideation in older people. PMID- 15877550 TI - Type and intensity of activity and risk of mobility limitation: the mediating role of muscle parameters. AB - OBJECTIVES: To investigate the association between different types of physical activity behavior and incident mobility limitation in older men and women and to examine whether muscle parameters mediate these associations. DESIGN: Cohort study with 4.5-year follow-up. SETTING: Metropolitan areas surrounding Pittsburgh, Pennsylvania, and Memphis, Tennessee. A random sample of white Medicare beneficiaries and all age-eligible blacks. PARTICIPANTS: Three thousand seventy-five black and white men and women aged 70 to 79 with no self-reported difficulty walking one-quarter of a mile or climbing 10 steps, enrolled in the Health, Aging and Body Composition (Health ABC) Study. MEASUREMENTS: Participants were classified as exercisers (reporting > or = 1,000 kcal/wk of exercise activity), lifestyle active (reporting < 1,000 kcal/wk of exercise activity and > or = 2,719 kcal/wk of total physical activity), or inactive (reporting < 1,000 kcal/wk of exercise activity and < 2,719 kcal/wk of total physical activity). The study outcome, incident mobility limitation, was defined as two consecutive, semiannual self-reports of any difficulty walking one quarter of a mile or climbing 10 steps. Thigh muscle area, thigh muscle attenuation (a marker of fat infiltration in muscle), appendicular lean soft tissue mass, and isokinetic knee extensor strength were examined as potential mediators. RESULTS: Over 4.5 years, 34.3% of men and 47.4% of women developed mobility limitation. Inactive persons had twice the risk of incident mobility limitation as exercisers (hazard ratio (HR)=2.08, 95% confidence interval (CI)=1.60-2.70, for men, HR=1.98, 95% CI=1.51 2.60, for women). Lifestyle-active men and women had an intermediate risk (HR=1.47 and 1.44, respectively). For the lifestyle active and inactive, absence of walking activity conferred an additional risk of mobility limitation. Muscle parameters did not mediate the relationship between physical activity and mobility limitation, except for knee extensor strength in men. CONCLUSION: Exercise and an active lifestyle that includes walking protect against mobility loss in older men and women. Activity effects on muscle parameters do not explain this association. PMID- 15877552 TI - Effect of race and sex on primary care physicians' diagnosis and treatment of late-life depression. AB - OBJECTIVES: To examine primary care physician (PCP) contributions toward racial and sex differences in the diagnosis and treatment of late-life depression. DESIGN: Survey using a computerized instrument incorporating video interviews and text, with volunteer PCPs randomly assigned to one of four standardized video vignettes of an elderly patient depicting late-life depression. Vignettes differed only in the patient/actor's race (white/African-American) or sex. SETTING: American Academy of Family Physicians meeting, San Diego, California, 2002. PARTICIPANTS: One hundred seventy-eight U.S.-practicing postresidency PCPs who were asked to participate in a clinical decision-making study. MEASUREMENTS: The computerized survey instrument assessed PCPs' diagnoses, first-line treatment and management recommendations, and judgment of personal characteristics/behaviors for the patients in the vignettes. RESULTS: Eighty-five percent of all PCPs correctly diagnosed the elderly patient(s) with major depression. There were no significant differences in the diagnosis of depression, treatment recommendations, or PCP assessment of most patient characteristics by the race or sex of the patient/actor in the vignette, but PCP characteristics, most notably the location of medical school training (U.S. vs international), affected the likelihood of a depression diagnosis and treatment recommendations. CONCLUSION: Given standardized symptom-pictures, PCPs are just as likely to diagnose and treat depression in African-American as in white older people, suggesting that bias based simply on apparent patient race is not a likely explanation for the lower rates of depression diagnosis and treatment in older African Americans. PCPs who have trained at international medical schools may benefit from targeted training initiatives on the diagnosis and treatment of late life depression. PMID- 15877553 TI - The effect of surgical and nonsurgical treatment on longitudinal outcomes of lumbar spinal stenosis over 10 years. AB - OBJECTIVES: To assess the relative effect of initial surgical and nonsurgical treatment on longitudinal outcomes of patients with lumbar spinal stenosis over a 10-year follow-up period. DESIGN: A prospective observational cohort study. SETTING: Enrollment from community-based specialist practices throughout Maine. PARTICIPANTS: One hundred forty-four patients with lumbar spinal stenosis who had at least one follow-up: 77 initially treated surgically and 67 initially treated nonsurgically. INTERVENTION: Initial surgical or nonsurgical treatment. MEASUREMENTS: Clinical data were obtained at baseline and outcomes followed at regular intervals over 10 years with mailed questionnaires including patient reported symptoms of back pain, leg symptoms, back-specific functional status, and satisfaction. Longitudinal data were analyzed using general linear mixed models. In addition to treatment (initial surgical or nonsurgical care), time period, and the interaction between treatment and time, the models included baseline score, patient age and sex, and a time-varying general health status score. The effects of these covariates in explaining differences between treatment groups were also examined. The effect of subsequent surgical procedures was assessed using different analysis strategies. RESULTS: The 10-year rate of subsequent surgical procedures was 23% and 38% for patients initially treated surgically and nonsurgically, respectively, and the overall 10-year survival rate was 69%. Patients undergoing initial surgical treatment had worse baseline symptoms and functional status than those initially treated nonsurgically. For all outcomes and at each time point, surgically treated patients reported greater improvement in symptoms and functional status and higher satisfaction scores, indicative of better outcomes, than nonsurgically treated patients. However, the relative magnitude of the benefit diminished over time such that the relative differences for low back pain and satisfaction were no longer significant over long-term follow-up (both P=.08 for treatment effect between 5 and 10 years after controlling for covariates). Regardless of initial treatment received, patients undergoing subsequent surgical procedures reported less improvement in outcomes over time than patients who did not undergo subsequent procedures, but the relative differences between treatment groups were similar in analyses that controlled for outcomes after subsequent procedures. CONCLUSION: After controlling for covariates, patients initially treated surgically demonstrated better outcomes on all measures than those initially treated nonsurgically. Although outcomes of initial surgical treatment remained superior over time, the relative benefit of surgery diminished in later years, especially for low back pain and satisfaction. Patients undergoing subsequent surgery had worse outcomes regardless of initial treatment received, but excluding them did not change overall treatment group comparisons. The analytical methods described may be helpful in the design and analysis of future studies comparing treatment outcomes for patients with lumbar spinal stenosis. PMID- 15877554 TI - Nighttime insomnia treatment and education for Alzheimer's disease: a randomized, controlled trial. AB - OBJECTIVES: To evaluate whether a comprehensive sleep education program (Nighttime Insomnia Treatment and Education for Alzheimer's Disease (NITE-AD)) could improve sleep in dementia patients living at home with their family caregivers. DESIGN: A randomized, controlled trial. PARTICIPANTS: Thirty-six community-dwelling patients with Alzheimer's disease (AD) and their family caregivers. INTERVENTION: All participants received written materials describing age- and dementia-related changes in sleep and standard principles of good sleep hygiene. Caregivers in active treatment (n=17) received specific recommendations about setting up and implementing a sleep hygiene program for the dementia patient and training in behavior management skills. Patients in active treatment were also instructed to walk daily and increase daytime light exposure with the use of a light box. Control subjects (n=19) received general dementia education and caregiver support. MEASUREMENTS: Primary sleep outcomes were derived for patients and caregivers from 1 week of sleep-wake activity measured at baseline, posttest (2 months), and 6-month follow-up using an Actillume wrist-movement recorder. Secondary patient outcomes included the Epworth Sleepiness Scale, the Cornell Depression Scale, and the Revised Memory and Behavior Problem Checklist. Caregiver self-reports included the Pittsburgh Sleep Quality Index and the Center for Epidemiological Study of Depression Scale. RESULTS: Patients participating in NITE-AD showed significantly greater (P<.05) posttest reductions in number of nighttime awakenings, total time awake at night, and depression, and increases in weekly exercise days than control subjects. At 6-month follow-up, treatment gains were maintained, and additional significant improvements in duration of night awakenings emerged. When cognitive level was controlled, NITE-AD patients had lower longitudinal ratings of daytime sleepiness than controls. There was a trend for control subjects to spend more time in bed at 6 months than NITE-AD patients. CONCLUSION: This study provides the first evidence that patients with AD who are experiencing sleep problems can benefit from behavioral techniques (specifically, sleep hygiene education, daily walking, and increased light exposure) that are known to improve sleep in nondemented, institutionalized older adults. PMID- 15877555 TI - Randomized, controlled trial of a nonpharmacological intervention to improve abnormal sleep/wake patterns in nursing home residents. AB - OBJECTIVES: Abnormal sleep/wake patterns are common in nursing home residents. Lifestyle and environmental factors likely contribute to these poor sleep patterns. The objective of this study was to test a multidimensional, nonpharmacological intervention to improve abnormal sleep/wake patterns in nursing home residents. DESIGN: Randomized, controlled trial. SETTING: Four nursing homes in the Los Angeles area. PARTICIPANTS: Residents were screened for excessive daytime sleeping (asleep > or = 15% of daytime observations) and nighttime sleep disruption (asleep < 80% of nighttime hours, according to wrist actigraphy). Four hundred ninety-two residents were screened; 339 had excessive daytime sleeping. Of these, 133 had nighttime sleep disruption and consented to participate; 120 completed baseline assessments, and 118 (77% female, mean age 86.9, 90% non-Hispanic white) were randomized to intervention versus usual care. INTERVENTION: Five consecutive days and nights of efforts to decrease daytime in bed time, 30 minutes or more of daily sunlight exposure, increased physical activity, structured bedtime routine, and efforts to decrease nighttime noise and light. MEASUREMENTS: Seventy-two consecutive hours of wrist actigraphy (nighttime sleep) and structured behavioral observations (daytime sleep and participation in social and physical activities and social conversation) at baseline and repeated at follow-up while the intervention or usual care condition was in place. RESULTS: The only effect on nighttime sleep was a modest decrease in mean duration of nighttime awakenings in intervention participants (10.6 minutes at baseline, 9.8 minutes at follow-up) versus an increase in controls (9.8 minutes at baseline, 13.8 minutes at follow-up) (F=4.27, P=.04). There were no significant effects on percentage of nighttime sleep or number of nighttime awakenings. There was a significant decrease in daytime sleeping in intervention participants (32% of daytime observations asleep at baseline, 21% at follow-up), with no change in controls (32% at baseline, 30% at follow-up; F=20.68, P<.001). Intervention participants had increased participation in social (F=22.42, P<.001) and physical (F=12.65, P=.001) activities and social conversation (F=5.04, P=.03). CONCLUSION: A multidimensional, nonpharmacological intervention into lifestyle and environmental factors that likely contribute to abnormal sleep/wake patterns in nursing home residents resulted in decreased daytime sleeping and increased participation in social and physical activities and social conversation. Nonpharmacological interventions should be considered in the management of abnormal sleep/wake patterns in nursing home residents. The main effect may be a significant decrease in daytime sleeping, which may translate to an improvement in quality of life. PMID- 15877556 TI - A pilot investigation of the short-term effects of an interdisciplinary intervention program on elderly patients with hip fracture in Taiwan. AB - OBJECTIVES: To evaluate an interdisciplinary intervention program for older people with hip fracture in Taiwan. DESIGN: Randomized experimental design. SETTING: A 3,800-bed medical center in northern Taiwan. PARTICIPANTS: Elderly patients with hip fracture (N=137) were randomly assigned to an experimental (n=68) or control (n=69) group. INTERVENTION: An interdisciplinary program of geriatric consultation, continuous rehabilitation, and discharge planning. MEASUREMENTS: Demographic and outcome variables were measured. Outcome variables included service utilization, clinical outcomes, self-care abilities, health related quality-of-life (HRQOL) outcomes, and depressive symptoms. RESULTS: Subjects in the experimental group improved significantly more than those in the control group in the following outcomes: ratio of hip flexion 1 month after discharge (P=.02), recovery of previous walking ability at 1 month (P=.04) and 3 months (P=.001) after discharge, and activities of daily living at 1 month (P=.01) and 2 months (P=.001) after discharge. Three months after discharge, the experimental group showed significant improvement in peak force of the fractured limb's quadriceps (P=.04) and the following health outcomes: bodily pain (P=.03), vitality (P<.001), mental health (P=.02), physical function (P<.001), and role physical (P=.006). They also had fewer depressive symptoms (P=.008) 3 months after discharge. CONCLUSION: This intervention program may benefit older people with hip fractures in Taiwan by improving their clinical outcomes, self-care abilities, and HRQOL and by decreasing depressive symptoms within 3 months after discharge. PMID- 15877557 TI - Timing of hospice referral and families' perceptions of services: are earlier hospice referrals better? AB - OBJECTIVES: To determine whether families of patients who enroll in hospice near the end of life believe that they receive less benefit from hospice services than families of patients who enroll earlier. DESIGN: Semistructured interviews at the time of hospice enrollment and 1 month after the patient's death. SETTING: This study was conducted at the Hospice of Lancaster County, Lancaster, PA, over a 12 month period spanning 2003-2004. PARTICIPANTS: First-time hospice admissions. MEASUREMENTS: Interviews assessed the anticipated helpfulness of six hospice services (enrollment interviews) and the actual helpfulness of the same services (follow-up interviews). RESULTS: Length of stay in hospice was associated with families' reports of the number of services provided (Spearman rho=0.34, P<.001), and with the mean helpfulness ratings for the services that were provided (Spearman rho=0.34, P<.001), although even with short lengths of stay, most families reported that they received the services that they anticipated and that those services were helpful. CONCLUSION: These results indicate that families feel they receive greater benefits from longer lengths of stay in hospice. Future efforts to define an "optimal" length of stay in hospice should consider patients' and families' perceptions of the benefits that hospice offers. PMID- 15877558 TI - Capsaicin troche for swallowing dysfunction in older people. AB - OBJECTIVES: To determine whether oral capsaicin troche supplementation with every meal upregulates the impairment of upper respiratory protective reflexes such as the swallowing reflex and the cough reflex. DESIGN: Randomized, controlled study with recruitment through nursing homes. SETTING: Sendai, Japan, from September 2002 through December 2003. PARTICIPANTS: Sixty-four participants in nursing homes with a mean age+/-standard deviation of 81.9+/-1.0 with stable physical status. INTERVENTION: Participants were randomly assigned to the program for the supplementation of capsaicin trochisci or placebo trochisci before every meal for 4 weeks. MEASUREMENTS: Assessment of individual latency time of the swallowing reflex (LTSR) and cough reflex sensitivity. RESULTS: Before the commencement of this study, there were no significant baseline differences in multiple parameters between the intervention group and control group. LTSR in participants in the intervention group was significantly shorter than in the control group (P<.05). The odds ratio (OR) of the shortening of the LTSR of more than 1 minute in the intervention group was 3.4 (95% confidence interval (CI)=1.1-10.4), compared with the control group (P=.03). In particular, daily capsaicin supplementation significantly increased the ratio of LTSR reduction at 4 weeks after the study to baseline LTSR in the high-risk group (baseline LTSR >6.0 seconds) compared with the low-risk group (baseline LTSR <3.0 seconds) and the intermediate group (3.0 seconds or = 75). More than 5,000 IGHI resources were identified, and 1,142 (22%) were advanced for screening. Of those, 246 (21%) were already peer reviewed, and 354 (31%) were advanced for peer review. Of those, 181 (51% or 20% of all those advanced for screening excluding those already peer reviewed) had a favorable assessment. Poor credibility and educational features contributed to this low quality. IGHI authored by national, federal, or academic organizations and targeted to higher academic levels was more likely to have a favorable assessment (P=0.01). Much publicly available IGHI is of suboptimal quality, particularly with regard to credibility and educational domains. Healthcare professionals need to be aware of how to locate high-quality IGHI and how to assess the quality of available IGHI. PMID- 15877571 TI - Assessment of geriatric information on the drug label for commonly prescribed drugs in older people. AB - Older people contribute to a significant portion of all prescription drug use and expenditures in the United States. Despite this, older people are often excluded from clinical trials examining the safety and efficacy of drugs. It is unclear to what extent drugs commonly used in older people contain information about prescribing in older people on their product labeling. The objective of this study was to determine the availability of pharmacokinetic, safety, and dosing information with reference to older people on product labels or package inserts (PIs) for commonly prescribed drugs in patients aged 65 and older. The top 50 oral drugs prescribed at the University of Pittsburgh Medical Center were identified through a computerized search of pharmacy records. PIs for these drugs were evaluated for information on elderly patients. Information on drug use in older people was found in 41 (82%) of the PIs. Drugs marketed after 1990 were more likely to contain information on geriatric use, with pharmacokinetic information being the most common type. Approximately 50% of the PIs contained precautionary statements for older people, but most did not provide any specific problems that might be encountered in this population. Only 28 (56%) PIs had dosing information available, and of these, just eight provided specific milligram recommendations for elderly patients. This analysis provides evidence that more should be done to improve the availability of prescribing information, particularly dosing and safety, on drug labels for older people. Additional regulatory action requiring this information to be included on drug product labels may be warranted. PMID- 15877572 TI - Exploring chronically ill seniors' attitudes about discussing death and postmortem medical procedures. AB - Proxy decisions about postmortem medical procedures must consider the dead patient's likely preferences. Ethnic- and sex-based attitudes surely underlie such preferences but lack sufficient characterization to guide decisions. Therefore, this exploratory study interviewed Mexican-American, Euroamerican, and African-American seniors in San Antonio, Texas, for their attitudes about discussing death before it occurs and about organ donation, autopsy, and practice on cadavers. A rigorous content analysis identified themes. Majority attitudes of an ethnic group or sex subgroup here may characterize the group generally. Attitudes about discussing death differed only by ethnic group. Mexican Americans and Euroamericans favored such discussions, but African Americans did not. Attitudes about the postmortem procedures differed by ethnic group and sex. Overall, Mexican Americans viewed the procedures most favorably; Euroamericans, less so; and African Americans, least so. Men and women differed further within ethnic groups. Mexican-American men and women split evenly over organ donation, the men expressed no majority preference about autopsies and the women agreed to them, and the men refused and the women agreed to practice on their cadavers. Euroamerican men expressed no majority preferences, but Euroamerican women agreed to organ donation, had no majority preference about autopsies, and refused practice on their cadavers. African-American men expressed no majority preferences, and African-American women expressed none about organ donation or autopsies but refused practice on their cadavers. If confirmed, these ethnic- and sex-based attitudes can help health professionals tailor postmortem care to individual patients' preferences. PMID- 15877575 TI - Chronic infection and frailty: surrogate markers, associations, and causality. PMID- 15877577 TI - Use of analgesics in elderly nursing home residents. PMID- 15877578 TI - Reactogenicity trends by age to influenza and pneumococcal vaccines. PMID- 15877579 TI - Recreational rehabilitation improved cognitive function in vascular dementia. PMID- 15877580 TI - New bases for percutaneous caval filter placement on site in geriatric institution in patients aged 75 and older. PMID- 15877581 TI - Do we need delirium units? PMID- 15877583 TI - Frailty judgment by hospital team members: degree of agreement and survival prediction. PMID- 15877584 TI - High vitamin B12 level: a strong predictor of mortality in elderly inpatients. PMID- 15877585 TI - Self-management is not related to lower demand for primary care in independent living elderly. PMID- 15877586 TI - Reliability and validity of the Medical Outcomes Study Social Support Survey (Chinese version). PMID- 15877587 TI - Geriatricians and functional outcomes. PMID- 15877590 TI - Cost-effectiveness of face-to-face smoking cessation interventions: a dynamic modeling study. AB - OBJECTIVES: To estimate the cost-effectiveness of five face-to-face smoking cessation interventions (i.e., minimal counseling by a general practitioner (GP) with, or without nicotine replacement therapy (NRT), intensive counseling with NRT, or bupropion, and telephone counseling) in terms of costs per quitter, costs per life-year gained, and costs per quality-adjusted life-year (QALY) gained. METHODS: Scenarios on increased implementation of smoking cessation interventions were compared with current practice in The Netherlands. One of the five interventions was implemented for a period of 1, 10, or 75 years reaching 25% of the smokers each year. A dynamic population model, the RIVM chronic disease model, was used to project future gains in life-years and QALYs, and savings of health-care costs from a decrease in the incidence of 11 smoking-related diseases over a time horizon of 75 years. This model allows the repetitive application of increased cessation rates to a population with a changing demographic and risk factor mix. Sensitivity analyses were performed for variations in costs, effects, time horizon, program size, and discount rates. RESULTS: Compared with current practice, minimal GP counseling was a dominant intervention, generating both gains in life-years and QALYs and savings that were higher than intervention costs. For the other interventions, incremental costs per QALY gained ranged from about 1100 per thousand for telephone counseling to 4900 per thousand for intensive counseling with nicotine patches or gum for implementation periods of 75 years. CONCLUSIONS: All five smoking cessation interventions were cost effective compared with current practice, and minimal GP counseling was even cost saving. PMID- 15877591 TI - Economic and utilization outcomes associated with choice of treatment for venous thromboembolism in hospitalized patients. AB - OBJECTIVES: Hospital administrative data were analyzed to assess treatment patterns, in-hospital mortality, rates of hemorrhagic events and thrombus propagation, utilization of health care resources, and hospital costs associated with various treatments during inpatient care for venous thromboembolism (VTE). STUDY DESIGN: Data from inpatient records were collected for deep venous thrombosis (DVT) and pulmonary embolism (PE) encounters at 132 US hospitals between January 1999 and December 2000. Patients receiving the most frequently employed treatments were compared with respect to demographics, related procedures and diagnostics, length of stay, adverse events, in-hospital mortality, and hospital costs. RESULTS: A total of 953 primary DVT and 3933 primary PE admissions were identified. Most admissions involved treatment with unfractionated heparin and vitamin K antagonist (UFH/VKA, 64.2% of admissions), followed by UFH with VKA and low-molecular-weight heparin (UFH/LMWH/VKA, 14.4%), and LMWH/VKA (12.9%). Compared with those treated with UFH/VKA, patients treated with LMWH/VKA experienced higher anticoagulant costs (dollar 540 vs. dollar 106), but lower total hospital costs (dollar 5198 vs. dollar 5977) and shorter lengths of stay (4.4 vs. 5.7 days for those without PE and 5.7 vs. 6.7 days for those with PE). CONCLUSIONS: UFH/VKA was the most common regimen used to treat VTE. In spite of its higher medication cost, however, treatment with LMWH/VKA was associated with significantly shorter hospital stays and lower total hospitalization costs, compared with UFH/VKA. PMID- 15877592 TI - Managing prices for hospital pharmaceuticals: a successful strategy for New Zealand? AB - OBJECTIVES: In 2002, as part of a National Hospital Pharmaceutical Strategy, the New Zealand (NZ) government agency PHARMAC commenced a 3-year period of negotiating prices for 90% of hospital pharmaceuticals on behalf of all NZ public hospitals. The present study was undertaken to determine the effects of this first year of "pooled procurement." METHODS: Using price changes and volume data for each of their top 150 pharmaceutical items, chief pharmacists at 11 public hospitals calculated projected cost savings for the financial year July 2003 to June 2004. Researchers calculated total projected savings for all 11 hospitals, and for three types of hospitals. Estimates of projected savings were made for all 29 major public hospitals by using savings per bed and savings per bed-day. A sensitivity analysis was undertaken. Items showing savings were categorized by using the Anatomical Therapeutic Chemical classification system. RESULTS: For the 11 hospitals, the top 150 items comprised 612 different items. Projected savings for 2003 to 2004 were NZ dollar 2,652,814, NZ dollar 658,984, and NZ dollar 127,952 for tertiary, secondary, and rural/special hospitals, respectively. Percentage savings as a median (range) of the total top 150 expenditure were: tertiary 5.28% (3.09-16.05%), secondary 7.41% (4.67-12.85%), and rural/special 9.55% (6.27-10.09%). For all 29 hospitals, estimated projected savings were NZ dollar 5,234,919 (NZ dollar 3,304,606-NZ dollar 8,044,482) by savings per bed, and NZ dollar 5,255,781 (NZ dollar 2,936,850-NZ dollar 8,693,239) by savings per bed-day. The main contributors to savings were: agents for infections, the nervous system, musculoskeletal system, and blood/blood-forming organs. CONCLUSION: The first year of pooled procurement under the National Hospital Pharmaceutical Strategy (2002-2003) has resulted in moderate savings. For all 29 major public hospitals, savings of around NZ dollar 5.2 million (dollar 2.9 million-dollar 8.7 million) or 3.7% were projected for 2003 to 2004. Longer-term effects, however, on patient outcomes and availability of pharmaceuticals, as well as on pharmaceutical expenditure, have yet to be evaluated. PMID- 15877593 TI - The economic value of childhood varicella vaccination in France and Germany. AB - OBJECTIVE: To determine the economic impact of childhood varicella vaccination in France and Germany. METHODS: A common methodology based on the use of a varicella transmission model was used for the two countries. Cost data (2002 per thousand) were derived from two previous studies. The analysis focused on a routine vaccination program for which three different coverage rates (CRs) were considered (90%, 70%, and 45%). Catch-up strategies were also analyzed. A societal perspective including both direct and indirect costs and a third-party payer perspective were considered (Social Security in France and Sickness Funds in Germany). RESULTS: A routine vaccination program has a clear positive impact on varicella-related morbidity in both countries. With a 90% CR, the number of varicella-related deaths was reduced by 87% in Germany and by 84% in France. In addition, with a CR of 90%, routine varicella vaccination induces savings in both countries from both societal (Germany 61%, France 60%) and third-party payer perspectives (Germany 51%, France 6.7%). For lower CRs, routine vaccination remains cost saving from a third-party payer perspective in Germany but not in France, where it is nevertheless cost-effective (cost per life-year gained of 6521 per thousand in the base case with a 45% CR). CONCLUSION: Considering the impact of vaccination on varicella morbidity and costs, a routine varicella vaccination program appears to be cost saving in Germany and France from both a societal and a third-party payer perspective. For France, routine varicella vaccination remains cost-effective in worst cases when a third-party payer perspective is adopted. Catch-up programs provide additional savings. PMID- 15877594 TI - Federal sponsorship of cost-effectiveness and related research in health care: 1997-2001. AB - OBJECTIVES: To describe recent federal sponsorship of cost-effectiveness and related health economics research to provide insight into the functioning of existing research support systems and assess the roles of federal health agencies. METHODS: Using the PubMed database, we identified cost-effectiveness and related publications citing support from a US government entity and published during the period of 1997 through 2001, and audited them for information on funding sources, study type, and content focus. RESULTS: Five Department of Health and Human Services agencies and centers and the Veterans Administration are cited as funders in 74% of 520 federally supported health economics publications we identified. Three-fourths of federally supported publications address five areas of high disease burden: infections, cancer, HIV/AIDS, cardiovascular disease, and substance abuse. Other high burden diseases, including mental health, diabetes, and injuries, receive less attention. Federal support of health economics studies of health education and care delivery intervention types underexamined in the field-is relatively strong but most often focuses on substance abuse or mental health services. Each of the top federal funders has a distinct funding pattern, but there are substantial areas of overlap within which we could not identify content domains specific to one funder or another. CONCLUSIONS: Federal support of health economics research has paralleled growth in the field. Federal funders support projects consistent with their mission and focus on high-burden disease areas. However, overlapping funding areas, ambiguity concerning agency interests within overlapping content areas, and gaps in some disease and intervention areas suggest that the coordination of health economics research funding could be improved. PMID- 15877596 TI - Potential roles for short RNAs in lymphocytes. AB - RNA interference (RNAi) is an ancient and evolutionarily conserved process. In some eukaryotes, RNAi silences parasitic genetic elements. In plants, RNAi serves as an immune system against RNA viruses and transgenes and in worms, RNAi silences transposons. In mammals, RNAi has yet unknown functions. However, emerging roles for short RNAs and the factors that interact with them in other eukaryotes include chromatin modification, DNA deletion and DNA methylation, which may provide clues to the roles for short RNA function in mammals. For example, antigen receptor expression in lymphocytes is a highly regulated process and although much is known about chromatin modification and DNA deletion in the immune system, several molecular details of chromatin regulation remain elusive. This review compares emerging roles for short RNA function to processes required for antigen receptor expression in mammalian lymphocytes and predicts that short RNAs direct events required for successful lymphocyte-restricted gene expression. PMID- 15877597 TI - Immune responses to dsRNA: implications for gene silencing technologies. AB - Nucleic acid-induced gene silencing, such as RNA interference (RNAi), induces a multitude of responses in addition to the knockdown of a gene. This is best understood in the context of the antiviral immune response, from which the processes of RNAi are thought to be derived. Viral challenge of a vertebrate host leads to an intricate series of responses that orchestrate antiviral immunity. The success of this multifaceted system in overcoming viral encounters hinges on complex pathogen-host interactions. One aspect of these interactions, the nucleic acid-based immune response, is key to the successful resolution of a viral challenge. In particular, dsRNA, a nucleic acid associated with viral replication, is involved in numerous interactions contributing to induction, activation and regulation of antiviral mechanisms. Specifically, dsRNA is responsible for stimulating important protective responses, such as the activation of dicer-related antiviral pathways, induction of type 1 IFN, and stimulation of dsRNA-activated protein kinase and oligoadenylate synthetase. Furthermore, the modulation and shaping of this overall immune response is facilitated through nucleic acid interactions with pattern recognition receptors such as toll-like receptor 3. These diverse dsRNA-induced antiviral responses have implications for biotechnologies that use dsRNA to harness one arm of the host antiviral machinery for silencing a specific target gene. The interlinked nature of these response elements means that it may be difficult to completely isolate one element from the other arms of the antiviral response program of an organism. Thus, it is beneficial to understand all aspects of the immune response to dsRNA in order to manipulate these systems and minimize unwanted non-specific effects. PMID- 15877598 TI - Genome-wide screening for gene function using RNAi in mammalian cells. AB - Mammalian genome sequencing has identified numerous genes requiring functional annotation. The discovery that dsRNA can direct gene-specific silencing in both model organisms and mammalian cells through RNA interference (RNAi) has provided a platform for dissecting the function of independent genes. The generation of large-scale RNAi libraries targeting all predicted genes within mouse, rat and human cells, combined with the large number of cell-based assays, provides a unique opportunity to perform high-throughput genetics in these complex cell systems. Many different formats exist for the generation of genome-wide RNAi libraries for use in mammalian cells. Furthermore, the use of these libraries in either genetic screens or genetic selections allows for the identification of known and novel genes involved in complex cellular phenotypes and biological processes, some of which underpin human disease. In this review, we examine genome-wide RNAi libraries used in model organisms and mammalian cells and provide examples of how these information rich reagents can be used for determining gene function, discovering novel therapeutic targets and dissecting signalling pathways, cellular processes and complex phenotypes. PMID- 15877599 TI - Detection of foreign RNA: implications for RNAi. AB - RNA interference (RNAi) is an exciting technology with applications in basic research, in elucidation and validation of drug targets, and as a direct therapeutic. In mammalian settings, it is based on the introduction or expression of small interfering RNA (siRNA) that guide the cleavage of a complementary target messenger RNA. While siRNA certainly directs specific silencing of genes in mammalian cells, longer RNA typically used to silence genes in other organisms potently activate mammalian cell defence mechanisms leading to a non-specific halt in translation, to activation of transcription and often, to cell death. Recent research has revealed that siRNA in certain settings can also activate these RNA-responsive pathways. With the recent advances in RNAi technology and its first forays into the in vivo setting now coming to light, it is pertinent to review the cellular response to ribonucleic acids typically used in RNAi methods. PMID- 15877600 TI - Transgenic plants for the production of veterinary vaccines. AB - The expression of antigens in transgenic plants has been increasingly used in the development of experimental vaccines, particularly oriented to the development of edible vaccines. Hence, this technology becomes highly suitable to express immunogenic proteins from pathogens. Foot and mouth disease virus, bovine rotavirus and bovine viral diarrhoea virus are considered to be the most important causative agents of economic loss of cattle production in Argentina, and they are thus optimal candidates for alternative means of immunization. Here, we present a review of our results corresponding to the expression of immunogenic proteins from these three viruses in alfalfa transgenic plants, and we discuss the possibility of using them for the development of plant-based vaccines. PMID- 15877601 TI - Is there a role for plant-made vaccines in the prevention of HIV/AIDS? AB - Although educational programs have had some impact, immunization against HIV will be necessary to control the AIDS pandemic. To be effective, vaccination will need to be accessible and affordable, directed against multiple antigens, and delivered in multiple doses. Plant-based vaccines that are heat-stable and easy to produce and administer are suited to this type of strategy. Pilot studies by a number of groups have demonstrated that plant viral expression systems can produce HIV antigens in quantities that are appropriate for use in vaccines. In addition, these plant-made HIV antigens have been shown to be immunogenic. However, given the need for potent cross-clade humoral and T-cell immunity for protection against HIV, and the uncertainty surrounding the efficacy of protein subunit vaccines, it is most likely that plant-made HIV vaccines will find their niche as booster immunizations in prime-boost vaccination schedules. PMID- 15877602 TI - The next 15 years: taking plant-made vaccines beyond proof of concept. AB - Significant potential advantages are associated with the production of vaccines in transgenic plants; however, no commercial product has emerged. An analysis of the strengths, weaknesses, opportunities and threats for plant-made vaccine technology is provided. The use of this technology for human vaccines will require significant investment and developmental efforts that cannot be supported entirely by the academic sector and is not currently supported financially by industry. A focus on downstream aspects to define potential products, conduct of additional basic clinical testing, and the incorporation of multidisciplinary strategic planning would accelerate the potential for commercialization in this field. Estimates of production cost per dose and volume of production are highly variable for a model vaccine produced in transgenic tomato, and can be influenced by the optimization of many factors. Commercialization of plant-made vaccine technology is likely to be led by the agricultural biotechnology sector rather than the pharmaceutical sector due to the disruptive nature of the technology and the complex intellectual property landscape. The next major milestones will be conduct of a phase II human clinical trial and demonstration of protection in humans. The achievement of these milestones would be accelerated by further basic investigation into mucosal immunity, the codevelopment of oral adjuvants, and the integration of quality control standards and good manufacturing practices for the production of preclinical and clinical batch materials. PMID- 15877603 TI - Oral hepatitis B vaccine candidates produced and delivered in plant material. AB - Hepatitis B is a major global health problem; approximately two billion people are infected with the virus worldwide, despite the fact that safe and efficacious vaccines have been developed and used for nearly 20 years. Prohibitive costs for vaccine purchase and administration restrict uptake in many developing nations. Agencies such as the Global Alliance for Vaccination and Immunization are helping to make current vaccines more available, but reduced costs would greatly aid this effort. Oral delivery is an option to reduce the expense of administering hepatitis B vaccines. It may also improve compliance, and orally delivered vaccines may be more efficacious among poor responders to current vaccines. However, to induce protective efficacy, oral administration may require encapsulation of antigen and delivery of large doses. Plant-based expression systems offer an oral delivery alternative with low production costs, and they also encapsulate the antigen. Some plant-based systems also stabilize antigen and therefore reduce storage and distribution costs. The hepatitis B major surface antigen has been expressed in several plant systems. A variety of regulatory sequences and subcellular targets have been used to achieve expression suitable for early stage clinical trials. However, further increase in expression will be necessary for practical and efficacious products. Appropriate processing can yield palatable products with uniform antigen concentration. The antigen expressed in plant systems shows extensive disulphide cross-linking and oligomerization and forms virus-like particles. Oral delivery of the antigen in plant material can induce a serum antibody response, prime the immune system for a subsequent injection of antigen and give a boosted response to a prior injection. Small scale clinical trials in which the antigen has been delivered orally in edible plant material indicate safety and immunogenicity. PMID- 15877604 TI - Use of viral vectors for vaccine production in plants. AB - The small size of plant viral genomes, the ease with which they can be manipulated, and the simplicity of the infection process is making the viral vectors an attractive alternative to the transgenic systems for the expression of foreign proteins in plants. One use of these virus expression systems is for vaccine production. There are two basic types of viral system that have been developed for the production of immunogenic peptides and proteins in plants: epitope presentation and polypeptide expression systems. In this review, we discuss advances made in this field. PMID- 15877605 TI - Expression systems and developments in plant-made vaccines. AB - Delivery of vaccines to mucosal surfaces can elicit humoral and cell-mediated responses of the mucosal and systemic immune systems, evoke less pain and discomfort than parenteral delivery, and eliminate needle-associated risks. Transgenic plants are an ideal means by which to produce oral vaccines, as the rigid walls of the plant cell protect antigenic proteins from the acidic environment of the stomach, enabling intact antigen to reach the gut associated lymphoid tissue. In the past few years, new techniques (such as chloroplast transformation and food processing) have improved antigen concentration in transgenic plants. In addition, adjuvants and targeting proteins have increased the immunogenicity of mucosally administered plant-made vaccines. These studies have moved plant-made vaccines closer to the development phase. PMID- 15877606 TI - Transfection of human monocyte-derived dendritic cells with CpG oligonucleotides. AB - Monocyte-derived dendritic cells (mDC), the most frequently applied DC subset in clinical studies, which can be obtained easily from peripheral blood monocytes after incubation with GM-CSF and IL-4, have not been clearly demonstrated to be activated by CpG oligodeoxynucleotides (ODN). The development of novel molecular strategies - such as the use of CpG-ODN - to increase immunological functions and thus improve the therapeutic efficiency of mDC vaccines in the treatment of malignant diseases is highly desirable. CpG-ODN need to be internalized into specific intracellular compartments to be active. Therefore, we applied electroporation and lipofection and compared these techniques with incubation to overcome possible defects in localization. Conditions of CpG-ODN transfection of these cells were optimized using fluorescein-marked ODN 2216. We were able to achieve high transfection efficiencies with various methods of delivery. However, we did not observe increased expression of maturation-associated and functionally relevant surface antigens (CD14, HLA-DR, CD40, CD83, CD80 and CD86), significant secretion of IL-12 and IFN-alpha in culture supernatant, or enhanced antitumour activation of cytokine-induced killer cells. In conclusion, our results show that non-viral transfection of CpG-ODN is not sufficient to overcome resistance of mDC to CpG activation. PMID- 15877607 TI - Upregulation of LPS-induced chemokine KC expression by 15-deoxy-delta12,14 prostaglandin J2 in mouse peritoneal macrophages. AB - 15-Deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) was initially identified as a high affinity natural ligand for the peroxisome proliferator-activated receptor (PPAR)-gamma. Recent studies have shown that it has a potent anti-inflammatory effect by attenuating the expression of proinflammatory mediators in activated macrophages, mainly through the inhibition of nuclear factor (NF)-kappaB dependent transcription of inflammatory genes. In this study, we investigated the synergistic effect of 15d-PGJ(2) on the expression of LPS-induced chemokine KC mRNA in mouse peritoneal macrophages. The time course of KC mRNA expression in cells stimulated with 15d-PGJ(2) plus LPS simultaneously (15d-PGJ(2)/LPS) showed similar patterns to the cells treated with LPS alone, and 15d-PGJ(2) had no effect on the stability of LPS-induced KC mRNA expression. Although NF-kappaB activity in cells treated with LPS was augmented by 15d-PGJ(2), pyrrolidone dithiocarbamate (PDTC) did not block the synergistic effect of 15d-PGJ(2) on LPS induced KC mRNA expression. However, the synergistic effect of 15d-PGJ(2) was markedly inhibited when the macrophages were treated with a inhibitor of the mitogen-activated protein kinase (MAPK) signalling pathway, 2'-amino-3' methoxyflavine (PD98059). Therefore, the mechanism of synergistic action of 15d PGJ(2) on the expression of LPS-induced KC mRNA in mouse peritoneal macrophages is possibly related to the MAPK signalling pathway, not to NF-kappaB activation. These data may contribute to unravelling some of the different mechanisms contrary to the anti-inflammatory effect of 15d-PGJ(2). PMID- 15877608 TI - A Staphylococcus aureus mouse keratitis topical infection model: cytokine balance in different strains of mice. AB - Staphylococcus is a leading cause of the potentially blinding disease microbial keratitis. Even with the use of antibiotic therapy, the host inflammatory response continues to damage the cornea, which may lead to blindness. Manipulation of the host response may help improve patient outcome from this devastating disease. We aim to understand the contribution of the host response to Staphylococcus aureus infection. A S. aureus keratitis mouse model was developed in both C57BL/6 and BALB/c mice using two different strains of S. aureus (8325-4 and Staph 38). Twenty-four hours postinfection, mice were killed and eyes were harvested for enumeration of bacteria, polymorphonuclear leucocytes, chemokines and cytokines. The laboratory strain 8325-4 was not as virulent as the clinical isolate Staph 38. In vitro data showed a 250-fold increase in invasion of human corneal epithelial cells by Staph 38 compared to 8325-4. BALB/c mice were susceptible to S. aureus infection whereas C57BL/6 mice were resistant. The resistant C57BL/6 mice were polarized towards a Th2 response, which may be protective for these mice. IL-4, IL-10 and IL-6 were elevated significantly in C57BL/6 mice infected with Staph 38 (P < 0.05). Macrophage inflammatory peptide (MIP)-2 was also significantly elevated in C57BL/6 mice (P < 0.001). The susceptible BALB/c mice had a muted cytokine response, which suggests that S. aureus might be 'walled off' during infection and might avoid host defences. IL-4, IL-10 and IL-6 cytokines may be protective during Gram-positive corneal infection and therefore may be useful for adjunct therapies in the treatment of this disease. PMID- 15877609 TI - Contribution of the cornea to cytokine levels in the whole eye induced during the early phase of Pseudomonas aeruginosa challenge. AB - Pseudomonas aeruginosa keratitis is one of the most destructive diseases of the cornea. The host response to this infection is critical to the outcome, and is regulated by cytokines produced in the ocular tissue. In this study, we assessed the relative contribution of the cytokines produced in the cornea to the inflammatory response of the whole eye to gain a better understanding of the inflammatory and regulatory processes in the ocular environment during localized corneal infection. C57BL/6 mice were challenged by topical application of P. aeruginosa to wounded corneas. Corneas and whole eyes were harvested 24 h post challenge and bacterial numbers, myeloperoxidase levels and the levels of cytokines known to be important in keratitis were determined. The site of production of IL-6 and KC in the retina was determined by in situ hybridization. Before infection, 90% of macrophage inflammatory protein (MIP)-2 and approximately 80% of all IFN-gamma and IL-10 produced constitutively in the eye was found outside the cornea. Twenty-four hours after infection, bacterial numbers, levels of myeloperoxidase, and levels of MIP-2 and IL-1 were not different, whether measured in cornea or whole eye. However, expression of IL-6, KC, IFN-gamma and IL-10 was significantly greater in whole eyes than in the corneas of infected eyes. The cells expressing IL-6 and KC in the retina were identified by in situ hybridization. This study indicates that during corneal inflammation, the response of the whole eye as well as the cornea needs to be considered. PMID- 15877610 TI - Direct processing and presentation of antigen from malaria sporozoites by professional antigen-presenting cells in the induction of CD8 T-cell responses. AB - Irradiated malaria sporozoites induce better protection than viable untreated sporozoites. We observed early differences between irradiated and viable untreated sporozoites in priming responses in vivo to a protective CD8 T-cell epitope, pb9, of the circumsporozoite protein of Plasmodium berghei. Sporozoites were processed for MHC class I presentation by dendritic cells (DC) to prime pb9 specific IFN-gamma-producing CD8 T cells. DC pulsed with untreated and irradiated sporozoites were similarly capable of priming central memory T-cell responses, detectable by the IFN-gamma cultured ELISPOT assay. However, irradiation significantly enhanced sporozoites' ability to prime effector T-cell responses detectable by the IFN-gammaex vivo ELISPOT assay. Irradiation also enhanced the ability of splenic APC to process and present sporozoites in order to re stimulate pb9-specific polyclonal and clonal T-cell responses. Sporozoites did not stimulate T cells in the absence of APC. Over-irradiation decreased the sporozoites' T-cell stimulating capacity in vitro at high parasite doses, which may indicate that an optimal irradiation dose is necessary to induce protective immunity by sporozoite inoculation. The induction of sporozoite-specific CD8 T cell responses without the need for liver stage infection identifies a potentially important mechanism in the development of pre-erythrocytic immunity. PMID- 15877612 TI - Catheter ablation of atrial fibrillation versus atrioventricular junction ablation plus pacing therapy for elderly patients with medically refractory paroxysmal atrial fibrillation. AB - BACKGROUND: Catheter ablation of atrial fibrillation (AF) has become another nonpharmacologic therapeutic option for medically refractory paroxysmal AF. Whether this method is better than atrioventricular (AV) junction ablation plus pacing therapy is unknown. The purpose of this study was to compare the very long term (longer than 4 years) clinical outcomes of the 2 methods in elderly patients (>65 years old) with medically refractory paroxysmal AF. METHODS: From January 1995 to December 2001, 71 elderly patients with medically refractory paroxysmal AF were included; group 1 included 32 patients with successful AV junction ablation plus pacing therapy and group 2, 37 patients with successful catheter ablation of AF. RESULTS: After a mean follow-up of more than 52 months, the AF was better controlled in the group 1 patients than group 2 (100% vs 81%, P = 0.013), however, they had a significantly higher incidence of persistent AF (69% vs 8%, P < 0.001) and heart failure (53% vs 24%, P = 0.001). Furthermore, the incidence of ischemic stroke and cardiac death was similar between the 2 groups. Compared with the preablation values, a significant increase in the NYHA functional class (1.7 +/- 0.9 vs 1.4 +/- 0.7, P = 0.01) and significant decrease in the left ventricular ejection fraction (44 +/- 8% vs 51 +/- 10%, P = 0.01) were noted in the group 1 patients, but not in the group 2 patients. CONCLUSIONS: Although AV junction ablation plus pacing therapy better controlled the AF in elderly patients with medically refractory paroxysmal AF, that method was associated with a higher incidence of persistent AF and heart failure than catheter ablation of AF in the very long-term follow-up. PMID- 15877613 TI - Non-pharmacological rate or rhythm control--it is time for randomized studies. PMID- 15877614 TI - Relationship between successful ablation sites and the scar border zone defined by substrate mapping for ventricular tachycardia post-myocardial infarction. AB - INTRODUCTION: It is unknown if identification of scar border zones by electroanatomical mapping correlates with successful ablation sites determined from mapping during ventricular tachycardia (VT) post-myocardial infarction (MI). We sought to assess the relationship between successful ablation sites of hemodynamically stable post-MI VTs determined by mapping during VT with the scar border zone defined in sinus rhythm. METHODS AND RESULTS: Forty-six patients presenting with hemodynamically stable, mappable monomorphic VT post-MI and who had at least one such VT successfully ablated were prospectively included in the study. In each patient, VT was ablated by targeting regions during VT that exhibited early activation, +/- isolated mid-diastolic potentials, and concealed entrainment suggesting a critical isthmus site. Prior to ablation, a detailed sinus-rhythm CARTO voltage map of the left ventricle was obtained. A voltage <0.5 mV defined dense scar. Successful VT ablation sites were registered on the sinus voltage map to assess their relationship to the scar border zone. Of the 86 VTs, 68% were successfully ablated at sites in the endocardial border zone. The remaining VTs had ablation sites within the scar in (18%), in normal myocardium (4%), and on the epicardial surface (10%). There were no significant differences in VT recurrence amongst the different groups. CONCLUSION: Successful ablation sites of hemodynamically stable, monomorphic VTs post-MI are often located in the scar border zone as defined by substrate voltage mapping. However, in a sizable minority, ablation sites are located within endocardial scar, epicardially, and even in normal myocardium. PMID- 15877615 TI - Further insights into appropriate target sites and outcomes of catheter ablation of ventricular tachycardia in post-infarction patients. PMID- 15877616 TI - Increased intensity of anticoagulation may reduce risk of thrombus during atrial fibrillation ablation procedures in patients with spontaneous echo contrast. AB - INTRODUCTION: A 10% incidence of left atrial (LA) thrombus formation has been detected using intracardiac echocardiography (ICE) imaging monitoring during LA ablation for atrial fibrillation (AF). The aim of this study was to determine if the intensity of anticoagulation reduces LA thrombus formation during pulmonary vein isolation procedure in patients with AF and spontaneous echo contrast (SEC). METHODS AND RESULTS: We studied 511 patients (age 56 +/- 10 years) undergoing pulmonary vein ostial isolation/ablation using radiofrequency energy. SEC was detected in 179 of 511 patients with ICE imaging before dual transseptal catheterization. All patients were anticoagulated with heparin to achieve activated clotting time (ACT) 250-300 seconds (group I) or >300 seconds (group II) confirmed at 30-minute intervals. SEC was detected in 49/294 (16.7%) patients in group I versus 130/217 (59.9%) in group II (P < 0.0001). LA thrombus was observed in 33/294 (11.2%) patients in group I versus 6/217 (2.8%) in group II (P < 0.05). For those patients with SEC, LA thrombus was observed in 22/49 (44.9%) in group I versus 2/61 (4.6%) in group II (P < 0.0001). There were no significant differences in age, number of unsuccessful drugs, persistent AF, left ventricular ejection fraction, and LA diameter between the two groups. No clinical embolic event was observed with withdrawal of LA thrombus to the RA. CONCLUSION: ICE diagnosed SEC before transseptal catheterization identifies an increased risk of LA thrombus. Increased intensity of heparin anticoagulation (ACT >300 seconds) during LA ablation for AF may prevent LA thrombus formation especially in patients with SEC. PMID- 15877617 TI - Long-term incidence of malignant ventricular arrhythmia and shock therapy in patients with primary defibrillator implantation does not differ from event rates in patients treated for survived cardiac arrest. AB - INTRODUCTION: Recent trials have demonstrated benefit of prophylactic defibrillator (ICD) implantation compared to conventional treatment in high-risk patients. However, many patients have rare or no sustained arrhythmias following implantation. Our study addresses the question, whether patients with prophylactic defibrillator implantation have a lower risk for life-threatening ventricular tachycardia (VT) or ventricular fibrillation (VF) compared to sudden cardiac death (SCD) survivors. METHODS AND RESULTS: Over 7 years we enrolled 245 patients. Occurrence of spontaneous sustained VT/VF resulting in adequate ICD treatment was the endpoint. Incidence, type, and treatment of sustained arrhythmia in 43 previously asymptomatic ICD recipients (group B) were compared to data of 202 survivors of imminent SCD (group A). All patients had severely impaired left ventricular ejection fraction (<45%). Group B patients had long runs (>6 cycles, <30 s) of VT during Holter monitoring and inducible sustained arrhythmia. Incidence of rapid VT and VF (cycle length <240 ms/heart rate >250 bpm) after 4 years (35% in both groups, P = ns) and adequate defibrillator therapies (57% vs 55%, P = ns) were similar in both groups after univariate and multivariate analysis. Cumulative mortality tended to be lower in group B compared to group A, but the difference was not statistically significant. CONCLUSION: During long-term follow-up, incidence of sustained rapid ventricular arrhythmia in prophylactically treated patients is as high as that of SCD survivors. Benefit from defibrillator implantation for primary prevention (group B) appears to be comparable to that for survived cardiac arrest (group A). PMID- 15877618 TI - The nature of the game. PMID- 15877619 TI - A novel missense mutation in the SCN5A gene associated with Brugada syndrome bidirectionally affecting blocking actions of antiarrhythmic drugs. AB - Brugada syndrome is an inherited cardiac disorder caused by mutations in the SCN5A gene encoding the cardiac sodium channel alpha subunit, which can lead ventricular fibrillation and sudden death. Inattentive use of antiarrhythmic drugs potentially triggers fatal cardiac arrhythmias through further reduction of sodium current (I(Na)). We studied the molecular mechanism underlying a case of Brugada syndrome that showed no response to a class Ic antiarrhythmic drug. Molecular genetic studies of a patient with Brugada syndrome identified a novel mutation in SCN5A, which causes substitution of serine for asparagine (N406S) in S6 of domain I (IS6). The provocation test with pilsicainide, a class Ic antiarrhythmic drug, failed to exacerbate ST-segment elevation in this case. Electrophysiological analyses of the N406S-mutant channel expressed together with the beta1 subunit in HEK293 cells showed that the voltage dependence of activation was positively shifted by 16 mV and that intermediate inactivation was enhanced. Whereas tonic block by pilsicainide was not changed in the N406S channel, use-dependent block by pilsicainide was almost completely abolished, consistent with the clinical findings of the negative provocation test. In contrast, the N406S channel showed stronger use-dependent block by quinidine than the wild-type channel. We demonstrate a novel Brugada mutation N406S, which is associated with the discordant effects on blocking actions of antiarrhythmic drugs as well as the multiple channel gating defects. We emphasis that an antiarrhythmic drug may exert unpredicted effects in patients with channel mutations. PMID- 15877620 TI - Biatrial and three-dimensional mapping of spontaneous atrial arrhythmias in patients with refractory atrial fibrillation. AB - INTRODUCTION: While atrial fibrillation (AF) initiation in the pulmonary veins has been well-studied, simultaneous biatrial and three-dimensional noncontact mapping (NCM) has not been performed. We hypothesized that these two techniques would provide novel information on triggers, initiation, and evolution of spontaneous AF and permit study of different AF populations. METHODS AND RESULTS: The origin of atrial premature beats (APBs), onset of spontaneous AF and its evolution were analyzed in 50 patients with AF in the presence or absence of structural heart disease (SHD) and in different AF presentations (group A: Persistent, group B: Paroxysmal). In 45 patients, spontaneous APBs in the right atrium (RA; n = 60) and left atrium (LA; n = 25) with similar regional distributions regardless of heart disease status were demonstrated. In total, 22 patients (44%) had > or =2 disparate regional origins. Biatrial regional foci were seen with equal frequency in patients with SHD (31%), without SHD (40%), in group A (32%), and in group B (36%). Biatrial mapping and NCM showed organized monomorphic atrial tachyarrhythmias arising in the RA (17), septum (17), or LA (21) and were classified as atrial flutter (RA = 34, LA = 8), macro-reentrant atrial tachycardia (RA = 1, LA = 3) or focal atrial tachycardia (RA = 2, LA = 7). Their regional distribution was more extensive in patients with SHD and persistent AF compared with patients without SHD or paroxysmal AF. Simultaneous biatrial tachycardias were observed only in group A patients and those with SHD. CONCLUSIONS: Simultaneous biatrial and NCM permits successful AF mapping in different AF populations and demonstrates a biatrial spectrum of spontaneous triggers and tachycardias. Organized monomorphic tachycardias with multiple unilateral or biatrial locations are commonly observed in human AF. Patients with heart disease or persistent AF have a more extensive distribution as well as simultaneous coexistence of multiple tachycardias during AF. PMID- 15877621 TI - Atrial fibrillation: can we cure it if we can't explain it? PMID- 15877622 TI - Cooled needle catheter ablation creates deeper and wider lesions than irrigated tip catheter ablation. AB - OBJECTIVES: To design and test a catheter that could create deeper ablation lesions. BACKGROUND: Endocardial radiofrequency (RF) ablation is unable to reliably create transmural ventricular lesions. We designed an intramural needle ablation catheter with an internally cooled 1.1-mm diameter straight needle that could be advanced up to 14 mm into the myocardium. The prototype catheter was compared with an irrigated tip ablation catheter. METHODS: Ablation lesions were created under general anesthesia in 14 male sheep (weight 44 +/- 7.3 kg) with fluoroscopic guidance. Each of the catheters was used to create two ablation lesions at randomly allocated positions within the left ventricle. The irrigation rate, target temperature, and maximum power were: 20 mL/min, 85 degrees C, 50 W for the intramural needle catheter and 20 mL/min, 50 degrees C, 50 W for the irrigated tip catheter, respectively. All ablations were performed for 2 minutes. After the last ablation, blue tetrazolium (12.5 mg/kg) was infused intravenously. The heart was removed via a left thoracotomy after monitoring the sheep for one hour. RESULTS: There was no evidence of cardiac tamponade in any sheep. The intramural needle catheter lesions were significantly wider (10.9 +/- 2.8 mm vs 10.1 +/- 2.4 mm, P = 0.01), deeper (9.6 +/- 2.0 mm vs 7.0 +/- 1.3 mm, P = 0.01), and more likely to be transmural (38% vs 0%, P = 0.03). CONCLUSIONS: Cooled intramural needle ablation creates lesions that are significantly deeper and wider than endocardial RF ablation using an irrigated tip catheter in sheep hearts. This technology may be useful in treating ventricular tachycardia resistant to conventional ablation techniques. PMID- 15877623 TI - Sex modulates the arrhythmogenic substrate in prepubertal rabbit hearts with Long QT 2. AB - Females have a greater susceptibility to Torsade de Pointes in congenital and drug-induced long QT syndrome (LQTS) that has been attributed to the modulation of ion channel expression by sex hormones. However, little is known regarding sex differences in pre-puberty, that is, before the surge of sexual hormones. In patients with congenital LQTS types 1 and 2, male children tend to have a greater occurrence of adverse events, especially in 10-15 year olds, than their female counterpart. To evaluate whether the rabbit model of drug-acquired LQTS exhibits similar age dependences, hearts of prepubertal rabbits were perfused, mapped optically to record action potentials (APs) and treated with an I(Kr) blocker, E4031 to elicit LQTS2. As expected, AP durations (APD) were significantly longer in female (n = 18) than male hearts (n = 10), at long cycle length. Surprisingly, E4031 (50-250 nM) induced a greater prolongation of APDs in male than in female hearts, and in both genders reversed the direction of repolarization (apex --> base to base --> apex), enhancing dispersions of repolarization. Furthermore, in male hearts, E4031 (0.5 microM) elicited early afterdepolarizations (EADs) that progressed to polymorphic ventricular tachycardia (PVT) (n = 7/10) and were interrupted by isoproterenol (40 nM) and prevented by propranolol (0.5-2.5 microM). In female hearts, E4031 (0.5 microM) produced marked prolongations of APDs yet few EADs with no progression to PVT (n = 16/18). Thus, sex differences are opposite in prepubertal versus adult rabbits with respect to E4031-induced APD prolongation, EADs and PVT, underscoring the fact that APD prolongation alone is insufficient to predict arrhythmia susceptibility. PMID- 15877624 TI - Gender-related differences in ventricular repolarization: beyond gonadal steroids. PMID- 15877625 TI - Method to predict isthmus location in ventricular tachycardia caused by reentry with a double-loop pattern. AB - BACKGROUND: During electrophysiologic study, induction and mapping of clinical reentrant ventricular tachycardia can be difficult. Hence, analysis of sinus rhythm electrograms for reentry localization is of potential clinical relevance. Herein is described a method of sinus-rhythm electrogram shape analysis, that does not require arbitrary threshold values, for localization of double-loop reentrant circuits that drive clinical tachycardias. METHODS AND RESULTS: Reentrant ventricular tachycardia was induced by premature stimulation in 23 postinfarction canine hearts 4-5 days after left anterior descending (LAD) ligation. Sinus-rhythm activation maps were constructed from bipolar electrograms acquired at 196-312 sites in the epicardial border zone. The timing of all electrogram peak deflections during one cycle of sinus rhythm was determined, and the mean (MPD) and deviation (DPD) were taken, respectively, as estimates of the time of activation wavefront crossing, and the duration of local electrical activity. These variables were then mapped on a computerized grid. The line of most uniform and sharp MPD gradient predicted the propagation direction through the double-loop reentrant circuit isthmus that would occur upon tachycardia induction. The sharpest transitions in DPD bounding this line predicted the positions of arcs of block that would border the reentrant circuit isthmus during tachycardia. The actual and estimated isthmuses overlapped by a mean of 84.1 +/- 3.8%. In six experiments lacking inducible reentrant tachycardia, no line of sharp MPD gradient was present in the MPD maps. CONCLUSIONS: Analysis of multiple sinus-rhythm deflections can localize the reentrant ventricular tachycardia isthmus without introduction of arbitrary threshold points and peak choices that may lead to error. PMID- 15877626 TI - The antiarrhythmic peptide analog ZP123 prevents atrial conduction slowing during metabolic stress. AB - OBJECTIVE: As atrial conduction slowing is important in the pathogenesis of atrial reentry arrhythmias, a drug that increases atrial conduction or prevents atrial conduction slowing could serve to prevent atrial reentry arrhythmias. In this study, we investigated whether the novel stable antiarrhythmic peptide analog, ZP123, was able to prevent atrial conduction slowing. METHODS: We examined the effect of ZP123 on metabolic stress-induced changes in conduction velocity (CV) and on dynamic CV restitution in isolated left atria from male Sprague-Dawley rats. We performed binding of ZP123 to a broad panel of 80 different cardiac and noncardiac ion channels and receptors and examined the effect of ZP123 on HERG channel conductance. RESULTS: ZP123 dose-dependently prevented metabolic stress-induced atrial CV slowing at doses ranging from 1 nM to 10 microM. ZP123 did not affect CV during physiological conditions nor did it affect dynamic CV restitution. ZP123 had no effect on atrial contractility. ZP123 showed no or low affinity binding to all ion channels and receptors examined. ZP123 had no effects on HERG channel activity in concentrations that affected atrial conduction. The concentration of ZP123 giving maximal effect on atrial conduction (100 nM) inhibited the outward K(+)-current by 2.7 +/- 0.1%. CONCLUSION: ZP123 has no effects on atrial conduction during physiological conditions, but it selectively prevents atrial conduction slowing during metabolic stress. PMID- 15877627 TI - An unusual accessory pathway: anteroseptal to ventricular outflow region connection. AB - Accessory pathways are typically located along the left or right atrioventricular junction. Distinct ventricular pre-excitation patterns determined by surface electrocardiography can provide reasonable pathway localization prior to invasive mapping and catheter ablation. We report an accessory pathway producing an unusual electrocardiographic appearance suggestive of ventricular outflow region pre-excitation. Pacing maneuvers and standard intracardiac recordings confirmed an atrial insertion immediately adjacent to the atrioventricular (AV) node and supported a ventricular insertion at the outflow tract region well away from the AV junction. The elimination of pathway conduction was achieved with radiofrequency (RF) energy at the atrial insertion after successful ice mapping excluded AV block at the target site. This is the second such pathway observed at our institution over a 20-year period. PMID- 15877628 TI - From genes to cell therapy: molecular medicine meets clinical EP. PMID- 15877629 TI - Is atrial fibrillation a genetic disease? AB - Atrial fibrillation remains one of the most challenging arrhythmias for the clinician and basic researcher. Different approaches have been undertaken to improve its understanding; from the development of animal models to the analysis of genetic backgrounds in individuals with familial and acquired forms of the disease. In the last few years, a large body of evidence has shown that alterations in ionic currents are involved in the disease. However, it has not been until recently, with the genetic link between mutations in proteins responsible for these ionic currents and the familial disease, that we have been given the final evidence that atrial fibrillation can also be primarily an ion channelopathy. Despite the limited prevalence of the inherited diseases, it has been shown before that the knowledge gained in their study will be helpful in dealing with the most common acquired forms of the disease. Therefore, as data keep unraveling, clinicians can expect that soon better therapeutic and preventive options for atrial fibrillation will emerge from basic science. PMID- 15877630 TI - A case of junctional extrasystoles? PMID- 15877631 TI - Optical coherence tomography imaging of the purkinje network. PMID- 15877633 TI - Changes in sleep problems, parents distress and impact of sleep problems from infancy to preschool age for referred and unreferred children. AB - This article compares and describes changes in sleep problems in 3- to 5-year-old Icelandic children referred and unreferred for sleep problems in infancy and explores changes in parents' distress and the impact of children's sleep problems on families over time. The sample consisted of a clinical group (n = 31) that had been referred to a sleep-disorder clinic in infancy, and a comparative group (n = 150) of age-matched unreferred community children. Self-report scales assessed infant/child sleep pattern, the impact of the sleep problem on family life and parents' distress. Results showed that about half the children in the community group have had a sleep problem in infancy. Nightwakings improved in both the referred and unreferred group over time but remained more frequent in the unreferred group. The referred group had significantly more settling problems in infancy than the other group but settling improved markedly over time. Parents of referred children were more fatigued compared with others despite improvement of children's sleep problems over time. Mothers of referred children were however, less likely to perceive the sleep problem as troublesome for family life than the others. It is concluded that parents of referred children are more fatigued than parents of unreferred children and nightwakings are more likely to persist in children who had sleep problems in infancy than in those with no such problems. PMID- 15877634 TI - Adolescents' perception of the quality of orthodontic treatment. AB - The aim was to describe quality of care from a patient perspective among adolescents receiving orthodontic treatment and to assess the relationship between quality of care and outcome-related aspects. The research design was cross-sectional. The sample consisted of 151 young people (mean age 17.1 years, SD: 2.2; 53% girls and 47% boys) receiving orthodontic treatment in the Stockholm region in Sweden (response rate 75%). Data were collected using the Quality from the Patient's Perspective questionnaire. The highest quality of care perceptions were noted on items dealing with receiving the best possible orthodontic treatment and being treated with respect. Less favourable perceptions of the quality of care were found regarding the opportunity to participate in the decisions related to the orthodontic treatment. In order to improve the quality of care a more active involvement of these patients in the decision-making process is suggested. The item 'I received the best possible orthodontic treatment' noted the highest subjective importance rating. The youngest participants reported the most favourable scores and the oldest the least. The majority (74%) reported that they were 'completely satisfied' with the result of the orthodontic treatment. However, 52% claimed that they had not followed all of the advice obtained during the treatment period, and 29% indicated some or more hesitation about attending the same dentist for future treatment. PMID- 15877635 TI - Assessing health-related quality of life in adolescents: some psychometric properties of the first Norwegian version of KINDL. AB - The study presented in this paper is part of a larger Norwegian investigation among adolescents, where the overall aim is to develop methods to promote their quality of life (QoL), to discover risk factors or threats to adolescents' well being, and finally to prevent the negative effects of such factors. An adequate generic health-related quality of life (HR-QoL) measure is therefore needed. However, only a limited number of well validated instruments that measure HR-QoL in adolescents exist, and to date only a few has been translated into Norwegian. The purpose of this study was therefore to examine some psychometric properties of the first Norwegian version of a simple, generic, German HR-QoL questionnaire for adolescents, KINDL. The instrument consists of 24-items, distributed in six subscales, which correspond to six domains of adolescents' HR-QoL. Based on a sample of 239 healthy adolescents, the internal consistency reliability is satisfactory for both the total scale and the subscales of 'Self-esteem and Family', fairly good for the 'Emotional' subscale, but lower for the subscales 'Physical', 'Friends' and 'School'. Factor analyses, which concerns construct validity, yielded interpretable solutions. The factor solutions at item level were interpreted to be in line with the original subscales, while factor analysis at subscale level indicated that a common QoL core is involved. To conclude, the Norwegian version of KINDL appears, in general, to be a psychometrically acceptable method of measuring HR-QoL in healthy adolescents. However, the alpha values of some of the subscales are not optimal, and these scales should be used with caution in research and profession. Still KINDL-N is considered suitable for screening purposes in the public health area and especially within school-health care. PMID- 15877636 TI - Descriptions of pain in elderly patients following orthopaedic surgery. AB - The aims of this study were to investigate what words elderly patients, who had undergone hip surgery, used to describe their experience of pain in spoken language and to compare these words with those used in the Short-Form McGill Pain Questionnaire (SF-MPQ) and Pain-O-Meter (POM). The study was carried out at two orthopaedic and two geriatric clinical departments at a large university hospital in Sweden. Altogether, 60 patients (mean age =77) who had undergone orthopaedic surgery took part in the study. A face-to-face interview was conducted with each patient on the second day after the operation. This was divided into two parts, one tape-recorded and semi-structured in character and one structured interview. The results show that a majority of the elderly patients who participated in this study verbally stated pain and spontaneously used a majority of the words found in the SF-MPQ and in the POM. The patients also used a number of additional words not found in the SF-MPQ or the POM. Among those patients who did not use any of the words in the SF-MPQ and the POM, the use of the three additional words 'stel' (stiff), 'hemsk' (awful) and 'rad(d)(sla)' (afraid/fear) were especially marked. The patients also combined the words with a negation to describe what pain was not. To achieve a more balanced and nuanced description of the patient's pain and to make it easier for the patients to talk about their pain, there is a need for access to a set of predefined words that describe pain from a more multidimensional perspective than just intensity. If the elderly patient is allowed, and finds it necessary, to use his/her own words to describe what pain is but also to describe what pain is not, by combining the words with a negation, then the risk of the patient being forced to choose words that do not fully correspond to their pain can be reduced. If so, pain scales such as the SF-MPQ and the POM can create a communicative bridge between the elderly patient and health care professionals in the pain evaluation process. PMID- 15877637 TI - Delirium is associated with poor rehabilitation outcome in elderly patients treated for femoral neck fractures. AB - The aim of this study was to describe risk factors for delirium and the impact of delirium on the rehabilitation outcome for patients operated for femoral neck fractures. Sixty-one patients, aged 70 years or older, consecutively admitted to the Department of Orthopaedic Surgery at Umea University Hospital, Sweden for femoral neck fractures were assessed and interviewed during hospitalization and at follow up 4 months after surgery. Delirium occurred in 38 (62%) patients and those who developed delirium were more often demented and/or depressed. Patients with delirium were longer hospitalized and they were more dependent in their activity of daily living (ADL) on discharge and after 4 months. They had poorer psychological well-being and more medical complications than the nondelirious. A large proportion of the patients who developed delirium did not regain their previous walking ability and could not return to their prefracture living accommodation. Delirium after hip fracture surgery is very common especially among patients with dementia or depression. This study shows that delirium has a serious impact on the rehabilitation outcome from both short- and long-term perspectives. Because delirium can be prevented and treated, it is important to improve the care of elderly patients with hip fractures. PMID- 15877638 TI - Applicability of the Nursing Interventions Classification to describe nursing. AB - The aim of this survey was to test the applicability of the Nursing Interventions Classification (NIC) system for use in a future nursing information system for documenting nursing in an electronic patient record in Iceland. Also, the aim was to test the translation of NIC into Icelandic. In order to be applicable to nursing NIC needs to be sensitive enough to describe the work nurses do, differentiate between specialities in nursing, and be understandable to nurses. A sample of 198 nurses was asked to identify how often they used each of 433 NIC nursing interventions. Of the 36 most frequently used interventions half are within the physiological domain. Core nursing interventions were different between specialities, e.g. Analgesic Administration had a high mean score in surgical nursing, and Health Education in primary health care. anova for the 27 classes in NIC showed significant differences (p < 0.01) by all nursing specialities except one, Crisis Management. A Tukey post hoc test showed how nursing specialities were reflected differently in the NIC domains, e.g. medical/surgical nursing in the Physiological: Basic Domain, but psychiatric nursing in the Behavioural Domain. Factor analysis of classes in NIC show good resemblance with the domains in NIC and the structure of the classification is strongly supported, except the Safety Domain. The results from this study indicate that nurses in the sample consider NIC to be applicable to describe nursing. The language is a powerful tool and is central in reflecting nursing practice as well as supporting the construct of knowledge. The translation of NIC to Icelandic is one step in many in preparing nurses to use a standardized language which can also be used in an electronic patient record. PMID- 15877639 TI - Stroke Specific Quality of Life Scale: Danish adaptation and a pilot study for testing psychometric properties. AB - BACKGROUND AND PURPOSE: Stroke has an effect on many aspects of quality of life (QoL) and therefore it is important to measure the magnitude of the impact. Items in the American version of the Stroke Specific Quality of Life (SS-QoL) Scale are developed with help from patients with stroke and include even language impairments. AIMS: To translate and culturally adapt the American SS-QoL Scale, version 2.0, a stroke-specific instrument measuring health-related QoL, into a Danish version, and to perform a pilot testing of the psychometric properties of the instrument. METHODS: The guidelines recommended by Guillemin et al. (J Clin Epidemiol, 46, 1993, 1417) were followed including two independent forward and one back translation, inclusion of laymen, here patients with stroke and their spouses, pretest, and examining selected items with a probing technique. Four couples were included in the pretest, 14 in the pilot study. The patients for pretest and pilot study were selected strategically with regard to interest for participation, experience as stroke survivors, sufficient capacity of language perception and cohabitant status. RESULTS: In the Danish version instructions were made more distinct and formal than in the original American version. Explanatory examples within the items were excluded. One new item adjusted for Danish conditions replaced one in the original version, and the response categories concerning personality were adjusted for cultural differences. The pilot study resulted in a more explicit formulation of the instructions to mark each item only once and on the same line as the item. CONCLUSIONS: A Danish version of SS-QoL version 2.0 has been developed and content as well as format has proved relevant and acceptable to patients with stroke including those with impaired language production. The next step is testing the psychometric properties of the instrument. PMID- 15877640 TI - Curtailing: handling the complexity of body care in people hospitalized with severe COPD. AB - Assisted personal body care (APBC) tends to be regarded as an unproblematic nursing activity with little professional challenge. For severely ill patient diagnosed with chronic obstructive pulmonary disease (COPD) daily bathing and washing is, however, a significant self-preserving activity that requires substantial efforts on the part of both patients and nurses. The aim of this study was to report on the complex pattern of APBC in hospitalized patients with severe COPD and highlight significant characteristics that should be considered in clinical practice in order to support patients' comfort and well-being. The study used a grounded theory design with a generative and constant comparative approach. The sample consisted of 12 cases of nurse-patient interaction, based on data from participant observation of sessions of APBC, measures of patient's perceived degree of breathlessness and individual interviews with patients and nurses after the sessions. Findings show that APBC in hospitalized patients with severe COPD is a complex integrated pattern of body care activities that can proceed with a greater or lesser degree of success. The main problem is how to keep the patient's breathing under control while optimizing comfort and well being. Curtailing addresses this problem. Curtailing is a subtle, purposeful balancing of protection from breathlessness and promotion of patients' present and future functional capacity in order to preserve their integrity. The idea that body care is a simple task belonging to patients' private daily lives may obscure the importance of determining a mutual nurse-patient agenda and the professional nurse responsibility may consequently be nebulous. Finding in this study will contribute to the development of a comprehensive and detailed understanding of the APBC and suggest the need for further investigation of the interaction perspective. PMID- 15877641 TI - Family caregivers' subjective experiences of satisfaction in dementia care: aspects of burden, subjective health and sense of coherence. AB - Family caregivers experience both positive and negative reactions in caregiving situations. There has been considerably less published about the positive aspects, however. The general aim of this study was to explore a previously developed instrument to study rewards gained by caregivers and to determine the factors associated with satisfaction in family members caring for patients with dementia living at home. The study group consisted of 153 such family members. Standardized interview schedules exploring different background characteristics, and instruments for assessment of the degree of dementia in the patients and the caregivers' total burden and degree of satisfaction were used. Factorial analysis of the Caregiver's Assessment of Satisfactions Index was performed and it became more specific for conditions of dementia when it was reduced. Stressors as measured by the Caregiver Burden scale and satisfaction can coexist and assess different aspects of the caregiver's situation. The caregiver can perceive both moderate burden and great satisfaction at the same time, and further studies may help to broaden our understanding of how we can reduce the degree of burden whilst increasing the sense of satisfaction. PMID- 15877642 TI - Recognition of client values as a basis for tailored care: the view of Dutch expert patients and family caregivers. AB - In the Netherlands confusion is signalled about the introduction of new care concepts like demand-oriented care. The aim of this article is to explore the phenomenon 'interaction aimed at care tailored to the client demand' as seen by expert clients: patients and their family caregivers. Focus interviews were held with expert patients and expert family caregivers of the 'Dutch Council of the Chronically ill and the Disabled'. Grounded theory methodology was used to analyse the results. Recognition by the professional of client values underlying their demand (uniqueness, comprehensiveness, continuity of life, fairness and autonomy) and underlying the care-relationship (equality, partnership and interdependence) emerged as central element within the interaction. Feelings of recognition with the client seem to reinforce autonomy, self-esteem and participation. Recognition was optimally felt in a dialogue. Four professional competencies could be identified related to recognition: attentiveness (ongoing actions to know and understand the patient); responsiveness (active, committed and responsible care guided by respect of patient identity); being a critical partner in care (giving and grounding professional opinion and discuss boundaries); being a developer of client competencies (facilitating and developing client participation within care). The findings offer possibilities to operationalize care concepts aimed at tailored care. Further research aimed at refining and testing the hypothesis developed is recommended. PMID- 15877643 TI - Interruption of agitation of platelet concentrates: effects on in vitro parameters. AB - BACKGROUND AND OBJECTIVES: When platelet concentrates (PCs) are shipped from one centre to another, they may remain unagitated for a considerable period of time. It was therefore our aim to study the effects of interruption of agitation on the in vitro parameters of PCs stored in platelet additive solutions. MATERIALS AND METHODS: In this multicentre study, PCs were prepared either by apheresis or from pooled buffy coats, paired to minimize donor-dependent differences, and aliquoted into 3 units with a 'low concentration' (approximately 1 x 10(9) platelets/ml; groups A, B and C) and 3 units with a 'high concentration' (approximately 2 x 10(9) platelets/ml; groups D, E and F). The final composition of the storage medium was 30% plasma and 70% additive solution in all PCs. Either PASIIIM or Composol was used as the additive solution. Agitation was interrupted for 2 days (between days 3 and 5, groups A and D), or for 4 days (between days 1 and 5, groups B and E), and continuous agitation served as the reference (groups C and F). A number of in vitro parameters were used for testing on days 1, 5 and 7. RESULTS: On day 7, reference units C and F in PASIIIM had significantly higher pH values than the study units in PASIIIM, but all retained a pH of > 6.5 at 37 degrees C. Hypotonic shock response (HSR) results were significantly lower in the high concentration/4-day interruption group (E) than in the other groups. The low concentration groups in PASIIIM, with agitation interrupted for either 2 days (group A) or 4 days (group B), did not have HSR values significantly different from the respective references. Study groups A, B, D and E in Composol, a solution lacking phosphate, had a pH of approximately 6.5 on day 7, which was significantly lower than that of the references and of the corresponding units in PASIIIM. The pH values were > 7.0 in reference groups C and F in Composol, not significantly different from those in PASIIIM. HSR values were also significantly lower in the Composol study groups. On the other hand, the reference Composol groups showed results similar to units in PASIIIM. CONCLUSIONS: PCs in PASIIIM additive solution with a platelet concentration of approximately 1 x 10(9)/ml can sustain 4 days without agitation. Phosphate may be of importance in maintaining good in vitro characteristics during interruption of agitation. PMID- 15877644 TI - Perioperative stimulation of erythropoiesis with intravenous iron and erythropoietin reduces transfusion requirements in patients with hip fracture. A prospective observational study. AB - BACKGROUND AND OBJECTIVES: Patients undergoing surgery for hip fracture (HF) often receive perioperative allogeneic blood transfusions (ABT) to avoid anaemia. However, concerns about the adverse effects of ABT have prompted the review of transfusion practice and the search for a safer treatment of perioperative anaemia. MATERIALS AND METHODS: We prospectively investigated the effect of a blood-saving protocol of perioperative iron sucrose (3 x 200 mg/48 h, intravenously) plus erythropoietin (1 x 40,000 IU, subcutaneously) if admission haemoglobin level < 130 g/l, on transfusion requirements and postoperative morbid mortality in patients with HF (group 2; n= 83). A parallel series of 41 HF patients admitted to another surgical unit within the same hospital served as the control group (group 1). Perioperative blood samples were taken for haematimetric, iron metabolism and inflammatory parameter determination. RESULTS: This blood-saving protocol reduced the number of transfused patients (P < 0.001), the number of transfused units (P < 0.0001), increased the reticulocyte count and improved iron metabolism. In addition, the blood-saving protocol also reduced the rate of postoperative infections (P = 0.016), but not the 30-day mortality rate or the mean length of hospital stay. CONCLUSIONS: The blood-saving protocol implemented seems to reduce ABT requirements in patients with HF, and is associated with a lower postoperative morbidity. The possible mechanisms involved in these effects are discussed. PMID- 15877645 TI - Does washing swabs increase the efficiency of red cell recovery by cell salvage in aortic surgery? AB - BACKGROUND AND OBJECTIVES: We investigated the contribution of swab washing to the efficiency of red cell recovery by intraoperative cell salvage (ICS) in 10 patients undergoing elective aortic aneurysm repair. MATERIALS AND METHODS: Volumes and haemoglobin (Hb) concentrations were recorded in the blood recovered by direct suction and from washed swabs, both before and after processing with a Haemonetics Cell Saver 5. RESULTS: The mean +/- standard deviation (SD) estimated blood loss was 991 +/- 403 ml, resulting in a mean +/- SD salvaged RBC volume of 380 +/- 124 ml. The median [interquartile (IQR) range] Hb collected from suction was 84.9 (61.8-131.4) g, of which 50.1 (45-71.5) g was returned to the patient after processing, a median yield of 68 (49-77)%. The swab wash produced a median (IQR) Hb of 39.4 (28.4-64.9) g, of which 26.2 (16.8-31) g was reinfused, a 67 (33 98)% yield. Swab wash thus contributed with a median (IQR) of 31 (24-39)% of the total RBC recovery. CONCLUSIONS: Washing swabs improves the efficiency of red cell recovery by ICS. PMID- 15877646 TI - Leukapheresis for the extraction of monocytes and various lymphocyte subpopulations from peripheral blood: product quality and prediction of the yield using different harvest procedures. AB - BACKGROUND AND OBJECTIVES: Leukapheresis of non-mobilized healthy donors is performed to harvest monocytes and lymphocyte subpopulations for use in various therapeutic regimens. In this methodological study, we compared two different leukapheresis programs, using equivalent volumes of processed blood over similar processing periods, to determine the influence of the procedures on the donor peripheral blood count and to establish the procedure that yields the highest quality product. MATERIALS AND METHODS: The target variables obtained in 41 healthy blood donors who underwent short-term leukapheresis (80-105 min) were retrospectively compared. Twenty-one volunteers were processed on a COBE Spectra machine at the MNC setting and 20 volunteers were processed at the AutoPBSC setting. Data were collected on pre- and postleukapheresis samples and on the product. RESULTS: AutoPBSC and MNC procedures resulted in a decrease of haemoglobin (5-7%), platelets (17-20%), monocytes (22%) and lymphocytes (23-27%), but not of granulocytes in peripheral blood. Both procedures produced nearly identical leucocyte and lymphocyte yields. AutoPBSC products contained a greater number of granulocytes, monocytes and red cells, but fewer platelets. The preleukapheresis values correlated with the yields for monocytes, T-helper and T suppressor cells, B-lymphocytes and natural killer cells, but not for granulocytes or platelets. CONCLUSIONS: Leukapheresis is a safe and efficient procedure for collecting large numbers of peripheral blood monocytes and different lymphocyte populations from non-mobilized donors. The two programs yield comparable leucocyte harvests. Based on our results, yields can be predicted from the peripheral cell counts. PMID- 15877647 TI - The Abantu phenotype in the ABO blood group system is due to a splice-site mutation in a hybrid between a new O1-like allelic lineage and the A2 allele. AB - BACKGROUND AND OBJECTIVES: Many phenotypic variations in the expression of blood group A have been explained by variations in gene structure, but unresolved samples are frequently encountered in the reference laboratory. Among ABO subgroups, A(bantu) has the highest frequency in a specified population. The molecular basis of this phenotype is now described. MATERIALS AND METHODS: Blood from Black donors phenotyped as A(bantu) was subjected to genomic ABO screening and direct sequencing of polymerase chain reaction (PCR)-amplified ABO exons 1-7 and introns 2-6. Total RNA was extracted and ABO cDNA was synthesized by reverse transcription (RT)-PCR. Control material comprised Black South African, Swedish, Jordanian and Brazilian blood samples with common phenotypes. RESULTS: Genomic ABO typing indicated the presence of an A(2) allele in each A(bantu) donor, in combination with an O allele. No previously reported mutations associated with weak A or B expression were found. Direct sequencing indicated the common A(2) sequence with a single nucleotide deletion (AGGT>AGT) at the exon 4/intron 4 junction, predicted either to disrupt the reading frame (resulting in a premature stop codon) or to cause erroneous splicing (resulting in the exclusion of exon 4 from the mRNA). O mRNA, but no transcripts from the A(bantu) allele, could be detected. Surprisingly, the splice-site mutation was also found in approximately 5% of O alleles in Black South Africans, but not in other blood donors, or in non O(1) alleles. Utilizing intron polymorphisms, the A(bantu) allele was shown to be a recombination between a new allelic lineage (O(1bantu)) and A(2), with a cross over region near exon 5. CONCLUSION: The A(bantu) phenotype is caused by an O(1bantu)-A(2) hybrid at the ABO locus. PMID- 15877648 TI - Blood group genotype analysis for the quality improvement of reagent test red blood cells. AB - BACKGROUND AND OBJECTIVES: Reagent red blood cells (RBCs) for antibody detection should express certain important antigens as a double dose, that is, the donors must be homozygous for the corresponding alleles. Traditionally, dose is determined by serological typing and known allele frequencies. However, RHD zygosity cannot be predicted serologically owing to the absence of an antithetical antigen, and FY zygosity is confounded by two variant haplotypes, FY*0 and FY*X. Furthermore, lack of reagents hampers our ability to type for some clinically important antigen pairs such as Do(a)/Do(b). MATERIALS AND METHODS: Genomic DNA was isolated from reagent RBC samples. Established, validated methods were used to determine the RHD, FY, and DO genotypes. RESULTS: Three of 52 D+ samples gave results that differed from the predicted genotype: two presumed R(1)R(1) samples and an R(2)R(2) sample were shown to be R(1)r' and R(2)r'', respectively. Five of 59 samples that were from presumed homozygotes for either FY*A or FY*B were heterozygous, together with either FY*X (three samples) or FY*0 (two samples). Seventy-five samples tested for DO were DO*A/A (n = 14), DO*A/B (n = 39), or DO*B/B (n = 22). CONCLUSIONS: The results show that serologically determined RhD and Duffy phenotypes of reagent RBCs are unreliable and that antigens we thought were represented as a double dose were single dose. The addition of Dombrock genotyping provides information which is useful in antibody identification. We conclude that selected genotype analyses are a valuable quality assurance measure to ensure that reagent RBCs comply with national and international recommendations for test sensitivity. PMID- 15877649 TI - Human platelet antigen genotyping by using sequence-specific primers and the particle gel agglutination assay. AB - BACKGROUND AND OBJECTIVES: Polymerase chain reaction using sequence-specific primers (PCR-SSP) is currently the most widely used technique for human platelet antigen (HPA) genotyping. Here, we describe a novel particle gel-agglutination technique for simplified visualization of the amplified products. MATERIALS AND METHODS: Biotinylated primers were used to amplify HPA-1, -2, -3, -4, -5, -6, and -15, and the PCR products were incubated with streptavidin particles. Fluorescein isothiocyanate (FITC)-labelled primers [amplifying a fragment of the human growth hormone (HGH) gene] and anti-FITC-coated particles were used as internal controls. Agglutination of the particles in or on top of the gel indicated specific amplification. A total of 100 samples from blood donors was tested by using this new technique and a standard PCR-SSP protocol. RESULTS: The use of biotinylated sequence-specific primers resulted in PCR products that agglutinated streptavidin particles, and the FITC-labelled HGH primers led to agglutination of anti-FITC-coated particles. Negative reactions were clearly distinguishable from positive reactions. The results of the particle gel agglutination method were in concordance with those of the electrophoretic visualization in all cases tested. CONCLUSIONS: The new particle agglutination method is reliable and easy to use. PMID- 15877650 TI - White blood cell fragments in platelet concentrates prepared by the platelet-rich plasma or buffy-coat methods. AB - BACKGROUND AND OBJECTIVES: White blood cell (WBC) fragments in platelet concentrates (PCs) may induce allo-immunization in the recipient. MATERIALS AND METHODS: As the level of WBC fragments can differ between PCs produced using different methods, we compared PCs prepared by using the buffy-coat method (BC PCs) in plasma or platelet additive solution (Composol) and PCs prepared using the platelet-rich plasma method (PRP-PCs). RESULTS: Post-filtration results revealed identical levels of WBC, but significantly higher CD62p expression and a significantly lower amount of total DNA, cell-free DNA and number of WBC fragments (0.6 vs. 4.2 WBC equivalents/microl) in PRP-PCs than in BC-PCs in either plasma or Composol. CONCLUSIONS: The number of WBC fragments is significantly higher in BC-PCs than in PRP-PCs. PMID- 15877651 TI - International collaborative study to evaluate a candidate reference preparation to define an appropriate specified limit of anti-D in intravenous immunoglobulin products. AB - BACKGROUND AND OBJECTIVES: The aim of the study was to evaluate a lyophilized intravenous immunoglobulin (IVIG) preparation containing anti-D (02/228; nominal reciprocal titre of 8) for its suitability to define the maximum limit of anti-D in IVIG products when used in a proposed reference method of direct haemagglutination of papain-treated erythrocytes, in an international collaborative study. MATERIALS AND METHODS: Twenty laboratories tested 02/228 along with a negative control IVIG preparation and four IVIG samples containing different levels of anti-D. Nineteen laboratories performed direct haemagglutination methodology using papain-treated erythrocytes; five of these laboratories and one additional laboratory performed their in-house haemagglutination methodology (all indirect antiglobulin tests). RESULTS: The mode titre of 02/228, obtained by using the proposed reference method, was 8 (62.5% of tests). However, there was wide variation in haemagglutination titres between laboratories for three of the four samples. Correcting the titres of the samples relative to those of the proposed reference preparation reduced the interlaboratory variability and increased the frequency of the mode titres in three out of four samples. The indirect antiglobulin tests also showed wide interlaboratory variability and were less sensitive than the direct method in four laboratories. Eleven of the 14 laboratories that expressed an opinion considered that the level of anti-D in 02/228 was appropriate to define a specified limit. CONCLUSIONS: Our results demonstrate the necessity of using a reference preparation to define the maximum level of anti-D in IVIG products and ensure sufficient sensitivity in haemagglutination testing methodology. On the basis of these results, members of the European Pharmacopoeia Expert Group 6B recommended revision of the appropriate monograph to include this new specification and test. The Food and Drug Administration in the USA intends to adopt the same maximal specification defined by the reference preparation and to recommend the same test for the safety of IVIG products. Preparations 02/228 and 02/226 were also established by the World Health Organization as International Reference Reagents to standardize haemagglutination testing for anti-D in normal IVIG products. PMID- 15877652 TI - Many prospective blood donors do not know for certain whether they have received blood transfusions. PMID- 15877653 TI - Implementation of donor screening for infectious agents transmitted by blood by nucleic acid technology: update to 2003. PMID- 15877658 TI - Medetomidine and dexmedetomidine: a review of cardiovascular effects and antinociceptive properties in the dog. AB - Alpha(2)-adrenoreceptor agonists (alpha(2)-agonists) are commonly used in small animal anaesthesia for their potent sedative and analgesic properties, although concerns about their cardiovascular effects have prevented their full adoption into veterinary practice. Research into alpha(2) adrenoreceptor agonists and their clinical use is extensive, therefore this review focuses on the use of dexmedetomidine and medetomidine in dogs. Emphasis is given to the cardiovascular effects and antinociceptive action of these agents. PMID- 15877659 TI - Electroencephalographic and cardiovascular variables as nociceptive indicators in isoflurane-anaesthetized horses. AB - OBJECTIVE: To evaluate Fourier-transformed electroencephalographic (EEG) variables, mean arterial blood pressure (MAP) and pulse rate as nociceptive indicators in isoflurane-anaesthetized horses. ANIMALS: Five standardbred and three Norwegian cold-blooded trotter stallions undergoing castration, aged 2-4 years, mass 378-538 kg. MATERIALS AND METHODS: All horses received intravenous (IV) detomidine (10 microg kg(-1) IV) and butorphanol (0.01 mg kg(-1) IV). Additional detomidine (4 microg kg(-1) IV) was administered in the induction area. Anaesthesia was induced with ketamine (2.5 mg kg(-1) IV) and diazepam (40 microg kg(-1) IV), and maintained for 30 minutes with isoflurane (end-tidal concentration of 1.4%) vaporized in oxygen. The electroencephalogram, MAP and pulse rate were recorded for 15 minutes, beginning 5 minutes before skin incision. Differences between the mean values of recordings taken before, and during surgery were calculated and tested for significant differences using a two sided Student's t-test. RESULTS: A significant rise in MAP and a fall in pulse rate were found. No significant change was found in any EEG variable. CONCLUSION/CLINICAL relevance Of the variables evaluated, MAP seems to be the most sensitive and reliable indicator of nociception in isoflurane-anaesthetized horses. PMID- 15877660 TI - The effects of spontaneous and mechanical ventilation on central cardiovascular function and peripheral perfusion during isoflurane anaesthesia in horses. AB - OBJECTIVE: To compare the effects of spontaneous breathing and mechanical ventilation on haemodynamic variables, including muscle and skin perfusion measured with laser Doppler flowmetery, in horses anaesthetized with isoflurane. STUDY DESIGN: Prospective controlled study. ANIMALS: Ten warm-blood trotter horses (five males, five females). Mean mass was 492 kg (range 420-584 kg) and mean age was 5 years (range 4-8 years). MATERIALS AND METHODS: After pre anaesthetic medication with detomidine (10 microg kg(-1)) anaesthesia was induced with intravenous (IV) guaifenesin and thiopental (4-5 mg kg(-1) IV) and maintained using isoflurane in oxygen. The horses were positioned in dorsal recumbency. In five animals breathing was initially spontaneous (SB) while the lungs of the other five were ventilated mechanically using intermittent positive pressure ventilation (IPPV). Total anaesthesia time was 4 hours with the ventilatory mode changed after 2 hours. During anaesthesia, heart rate (HR) cardiac output (Qt) stroke volume (SV) systemic arterial blood pressures (sAP), and pulmonary arterial pressure (pAP) were recorded. Peripheral perfusion was measured in the semimembranosus and gluteal muscles and on the tail skin using laser Doppler flowmetry. Arterial (a) and mixed venous (v) blood gases, pH, haemoglobin concentration [Hb], haematocrit (Hct), plasma lactate concentration and muscle temperature were measured. Oxygen content, venous admixture (s/Qt) oxygen delivery (DO(2)) and oxygen consumption (VO(2)) were calculated. RESULTS: During mechanical ventilation, HR, sAP, pAP, Qt, SV, Qs/Qt and PaCO(2) were lower and PaO(2) was higher compared with spontaneous breathing. There were no differences between the modes of ventilation in the level of perfusion, DO(2), VO(2), [Hb], (Hct), or plasma lactate concentration. After the change from IPPV to SB, left semimembranosus muscle and skin perfusion improved, while muscle perfusion tended to decrease when SB was changed to IPPV. Low-frequency flow motion was seen twice as frequently during IPPV compared with SB. CONCLUSIONS: Mechanical ventilation impaired cardiovascular function compared with SB in horses during isoflurane anaesthesia. Muscle and skin perfusion changes occurred with ventilation, although further studies are needed to elucidate the underlying mechanisms. PMID- 15877661 TI - Effect of medetomidine and its antagonism with atipamezole on stress-related hormones, metabolites, physiologic responses, sedation, and mechanical threshold in goats. AB - OBJECTIVE: To evaluate the effects of medetomidine and its antagonism with atipamezole in goats. STUDY DESIGN: Prospective randomized crossover study with 1 week between treatments. ANIMALS: Six healthy 3-year-old neutered goats (three male and three female) weighing 39.1-90.9 kg (60.0 +/- 18 kg, mean +/- SD). METHODS: Goats were given medetomidine (20 microg kg(-1), IV) followed, 25 minutes later, by either atipamezole (100 microg kg(-1), IV) or saline. Heart and respiratory rate, rectal temperature, indirect blood pressure, and mechanical threshold were measured, and sedation and posture were scored and blood samples obtained to measure epinephrine, norepinephrine, free fatty acids, glucose, and cortisol concentrations at baseline (immediately before medetomidine), 5 and 25 minutes after medetomidine administration, and at 5, 30, 60, and 120 minutes after the administration of antagonist or saline. Parametric and nonparametric tests were used to evaluate data; p < 0.05 was considered significant. RESULTS: Medetomidine decreased body temperature, heart rate, and respiratory rate and increased mean arterial blood pressure, cortisol, and glucose. Recumbency occurred 89 +/- 50 seconds after medetomidine administration. All goats were standing 86 +/- 24 seconds after atipamezole administration whereas all goats administered saline were sedate and recumbent at 2 hours. Tolerance to compression of the withers and metacarpus increased with medetomidine. From 5 to 120 minutes after saline or atipamezole administration, there were differences in body temperature, glucose, and cortisol but none in heart rate or blood pressure. Three of the six goats receiving saline developed bloat; five of six urinated. After atipamezole, four of six goats developed piloerection and all goats were agitated and vocalized. CONCLUSION: At the doses used, atipamezole antagonized the effects of medetomidine on recumbency, sedation, mechanical threshold, and the increase in glucose. Atipamezole increased the rate of return of cortisol toward baseline, and prevented further decline in rectal body temperature. CLINICAL RELEVANCE: Atipamezole may be used to antagonize some, but not all effects of medetomidine. PMID- 15877662 TI - Pre-operative stress in dogs - a preliminary investigation of behavior and heart rate variability in healthy hospitalized dogs. AB - OBJECTIVE: To assess pre-operative behavioral and physiological characteristics of healthy dogs hospitalized for elective surgery. STUDY DESIGN: Open clinical observational study. ANIMALS: Forty-one bitches hospitalized for elective ovariohysterectomy. METHODS: While undisturbed in a hospital cage, the behavior of the dog was recorded using a video camera and recorder. From the video recordings, various behavioral variables were registered. Simultaneous measurements were made on heart rates and heart rate variability (HRV) by use of an ambulatory electrocardiogram. In addition, the dog's response to human approach was noted. RESULTS: Different behavioral patterns were found in the dogs studied. Thirteen individuals were regarded as highly active, and were seen to bark or howl, manipulate the environment or attempt to flee vigorously. In 13 dogs (passive dogs) none of these activities occurred. Panting and displacement behaviors, such as snout licking, were observed in nearly all the animals monitored. In general, heart rates were higher and HRV lower with the most active individuals; however, the presence of physiological arousal could not be excluded in two animals with passive behaviors. To some extent, the behavior of the dog while undisturbed in the cage was reflected in the responses to a person entering the cage. CONCLUSIONS AND CLINICAL RELEVANCE: The different individual responses detected in this study raise an important question concerning their role and existence throughout the entire peri-operative period, especially during the post anesthetic recovery phase when behavioral characteristics are commonly used to assess patient welfare. The results also emphasize the need for further investigations to explore the effects of pre-operative stressors on canine surgical patients, and the factors contributing to them. PMID- 15877663 TI - Closed system anaesthesia in dogs using liquid sevoflurane injection; evaluation of the square-root-of-time model and the influence of CO2 absorbent. AB - OBJECTIVE: To determine whether predictable alveolar concentrations of sevoflurane are reliably produced in dogs when liquid sevoflurane is injected into closed circuit breathing systems, as calculated by Lowe's square-root-of time anaesthetic uptake model, and to confirm the validity of the model using soda lime and calcium hydroxide lime. STUDY DESIGN: Prospective clinical study. ANIMALS: Eleven healthy dogs with a mean body mass of 34 +/- 9 kg scheduled for pelvic limb orthopaedic surgery. MATERIALS AND METHODS: Following pre-anaesthetic medication, anaesthesia was induced with propofol and maintained with sevoflurane in a closed circle system. Epidural anaesthesia was performed with morphine and bupivacaine. Liquid sevoflurane was injected into the circuit by syringe, using dosages and time intervals derived from Lowe's square-root-of-time anaesthetic uptake model. The target alveolar concentration chosen was 1.1 x MAC (2.6% end tidal sevoflurane). Either soda lime (group S; n = 6) or calcium hydroxide lime (Amsorb; group A; n = 5) were used for CO(2) absorption. Sevoflurane concentration and the respiratory gas composition were measured with an infrared gas analyser. RESULTS: End-tidal sevoflurane concentrations were close to the predicted value of 2.6% at 9 minutes (2.53 +/- 0.1% group S; 2.60 +/- 0.26% group A) and 16 minutes (2.55 +/- 0.30 group S; 2.52 +/- 0.28% group A) but declined thereafter to reach 50% (group S) and 64% (group A) of the predicted value at 121 minutes. There was a constant trend towards higher end-tidal sevoflurane concentrations in group A but the difference was not statistically significant. CONCLUSIONS: The square-root-of-time model leads to significantly lower alveolar concentrations than expected, suggesting that the rate of sevoflurane uptake in dogs declines less rapidly than predicted. The use of Amsorb tends to reduce the deviation from predicted concentrations. CLINICAL RELEVANCE: The model used in this study provided only an approximate guide to the volume of liquid sevoflurane required. Consequently, the definitive dose schedule must be based on measured anaesthetic concentrations and clinical monitoring. PMID- 15877664 TI - Bone mineral density and urinary N-acetyl-beta-D-glucosaminidase activity in paediatric patients with idiopathic hypercalciuria. AB - BACKGROUND: Idiopathic hypercalciuria (IH) is defined as hypercalciuria that persists after correction of dietary inbalances and has no detectable causes. Patients with IH have a higher prevalence of osteoporosis. Defective reabsorption of calcium by the renal tubule is considered a likely mechanism of IH. N-acetyl beta-D-glucosaminidase (NAG) is a lysosomal enzyme that is a very sensitive marker of renal tubular impairment. METHODS: Fifteen patients (nine boys and six girls, mean age 12.4 +/- 4.0 years) with IH (urinary calcium excretion >0.1 mmol/kg per 24 h) had their bodyweight, height, body mass index (BMI), urinary NAG/creatinine ratio (U-NAG/Cr) and 24-h urinary calcium excretion (U-Ca/24 h) assessed. L1-L4 bone mineral density (BMD) was measured by dual energy X-ray absorptiometry and volumetric BMD (BMDvol) was calculated. The obtained results were expressed as Z-scores. RESULTS: The values of basic anthropometric parameters did not differ significantly from the values of the reference population and there was a tendency to short stature, which did not reach statistical significance (P = 0.08). The values of calciuria and U-NAG/Cr were significantly higher while BMD was significantly lower when compared to the reference values (P < 0.0006, P < 0.006 and P < 0.001, respectively). Inverse and significant correlations were found between U-Ca/24 h and BMD, U-Ca/24 h and body height, and U-Ca/24 h and BMDvol (r = -0.64 and -0.70, respectively, P < 0.01; r = -0.55, P < 0.05), while there was no correlation between U-NAG/Cr and U-Ca/24 h, nor between BMD and weight or BMD and BMI. CONCLUSION: Tubular impairment is highly probable in children with IH, but there is a poor relationship with the degree of calcium leakage. Idiopathic hypercalciuria should be considered as a risk factor for stunted growth and low bone mass. PMID- 15877665 TI - Acute interstitial nephritis due to mesalazine. AB - A case of mesalazine-induced acute interstitial nephritis (AIN) in a 41-year-old patient with ulcerative colitis (UC) is reported here. Clinical symptoms such as fever and arthralgia, and laboratory findings such as eosinophilia and renal failure suggested AIN, which was confirmed by biopsy. With withdrawal of mesalazine and intravenous methylprednisolone the patient's renal function was recovered. It is observed that early discontinuation of mesalazine is associated with amelioration of interstitial nephritis in most patients, so the recommendation is that patients receiving mesalazine should undergo routine monitoring of renal function. Delayed diagnosis may lead to permanent renal function impairment. PMID- 15877666 TI - Nephrology training in Australia: is there a problem? PMID- 15877667 TI - QT interval dispersion in dialysis patients. AB - The leading cause of mortality in dialysis patients is cardiovascular complications, including ventricular arrhythmias and sudden cardiac death. A reliable non-invasive predictive test of sudden death is therefore important. The interlead variation in duration of the QT interval on the surface electrocardiogram corrected with heart rate (QTc dispersion) might serve as a surrogate for ventricular arrhythmia. Prolonged QTc dispersion is commonly encountered in dialysis patients and possesses an increased risk of all mortality, including cardiovascular mortality. QT dispersion might be affected by shifts of the intracellular electrolytes during dialysis and increasing deposition of iron in cardiac muscles in these patients who have underlying heart diseases. Although no well-designed study has been done, the factors contributing to prolongation of QTc dispersion should be avoided. We summarize the results of the currently available clinical studies that examined QTc dispersion in dialysis patients. PMID- 15877668 TI - QTc dispersion in haemodialysis patients with cardiac complications. AB - BACKGROUND AND AIMS: QTc dispersion (Maximum - Minimum QTc interval on a standard 12-lead electrocardiogram (ECG)) is a useful predictor of tachyarrhythmic events and related sudden cardiac death. Recent studies have reported that QTc dispersion is prolonged in patients receiving haemodialysis (HD), and it is often further prolonged following HD treatment. In the present study, we examine whether the patients who are susceptible to prolongation of QTc dispersion by HD are prone to life-threatening cardiovascular events and related deaths. METHODS: Forty-eight patients with cardiac symptoms such as chest pain (n = 32), arrhythmia (n = 2) or heart failure (n = 14) and receiving chronic HD treatment (40 males and eight females) were subjected to the present study. Twenty-six of a total 48 patients were given a diagnosis of ischaemic heart disease (IHD) by coronary angiography. Ten patients had cerebrovascular accidents and 11 patients had peripheral vascular disease. After the ECG recording, a prospective follow up was conducted for 37 +/- 24 months. RESULTS: During the follow-up period, 30 patients had cardiac events, and 23 including 18 cardiovascular deaths occurred as outcomes. Throughout the total period, patients who showed prolongation of QTc dispersion after HD had cardiovascular deaths with higher incidence than those did not show the prolongation. CONCLUSIONS: We consider that prolongation of QTc dispersion after HD treatment can predict the prognosis of patients with renal failure bearing cardiac complications. Great care is necessary for such patients if they show longer QTc dispersion and/or susceptibility for further prolongation after HD treatment. PMID- 15877669 TI - Impact of haemodialysis on QTc dispersion in children. AB - BACKGROUND: The purpose of the present paper was to investigate the corrected QT (QTc) interval and QTc dispersion value, and the impact of haemodialysis on these parameters in children with chronic renal failure. METHODS: Nineteen patients with chronic renal failure receiving haemodialysis were included in the present study. Electrocardiography (ECG), echocardiography and serum biochemistry were performed in all patients. Serum electrolyte levels were measured before and after haemodialysis, at the time of the ECG. Nineteen healthy age- and sex matched children served as the control group. RESULTS: Patients with chronic renal failure had greater QTc interval and QTc dispersion compared to control subjects. The patients' sex, age and presence of hypertension or left ventricular hypertrophy (LVH) were not related to QTc interval/dispersion. However, the patients with left ventricular (LV) systolic dysfunction had significantly greater QTc dispersion value. After haemodialysis session, both QTc interval and QTc dispersion values significantly increased. Serum potassium levels significantly decreased, whereas the calcium level significantly increased after the haemodialysis session. The changes in electrolyte values were not associated with the changes in both QTc interval and QTc dispersion. CONCLUSION: Children receiving haemodialysis may be at greater risk of ventricular arrhythmia and sudden death because QTc dispersion reflects heterogeneous recovery of ventricular excitability. PMID- 15877670 TI - Effect of intravenous iron on haemodialysis catheter microbial colonization and blood-borne infection. AB - BACKGROUND: Intravenous (i.v.) iron is employed to treat absolute and relative iron deficiency in end-stage renal disease patients. However, there exists the possibility that i.v. iron increases infection risk. This pilot study examines whether i.v. iron gluconate acutely increases tunnelled haemodialysis catheter colonization, microbial growth, or blood-borne infection. METHODS: Nineteen patients with haemodialysis catheters who met criteria to receive an i.v. iron load entered the study. Six matched patients with catheters who did not receive iron were controls. Blood aspirated from the catheter prior to initiation of haemodialysis was sent for qualitative/quantitative cultures. The study consisted of three baseline cultures, five cultures during iron (125 mg of ferric gluconate per treatment), and three cultures following iron administration. Patients were monitored for infection for 30 days following iron. RESULTS: Fifteen iron-treated patients and six controls completed the study. Thirty-three per cent of treated patients were colonized at baseline; 66% were colonized following iron. Thirty three per cent of controls (2/6) were colonized at baseline; no new colonization developed during follow up. Neither treated patients nor controls had significant microbial growth within catheters; one patient in the iron-treated group developed candidaemia. CONCLUSION: Intravenous iron is not associated with acute microbial growth in catheters or clinical infection. However, a trend towards increased catheter colonization following iron administration exists. PMID- 15877671 TI - Randomized cross-over comparison of intravenous and subcutaneous darbepoetin dosing efficiency in haemodialysis patients. AB - BACKGROUND: Studies have consistently shown the superior dosing efficiency of subcutaneous (s.c.) compared to intravenous (i.v.) erythropoietin (r-HuEPO). Unlike r-HuEPO, data from pivotal darbepoetin trials support s.c. and i.v. dosing equivalence, however, no blinded cross-over randomized studies of s.c. and i.v. dose efficiency or intra-patient variability in response have been published. METHODS: During this 12-month study, 53 haemodialysis patients were randomized to s.c. or i.v. darbepoetin for a 6-month period and then switched to the alternative route for a second 6-month period. Darbepoetin dose was titrated during the first 4-months of each period to achieve a stable haemoglobin during the final 2-month observation period of each arm. RESULTS: Twenty-four patients were included in analysis. No significant difference between s.c. and i.v. administration was observed for any measured parameter. Patients achieved a non significantly higher haemoglobin (123.6 +/- 3.76 vs 120.9 +/- 4.42 g/L, P = 0.11) from a non-significantly lower darbepoetin dose (40.8 +/- 10.7 vs 42.5 +/- 11.0 mcg/week, P = 0.23) with i.v. administration. The population-based weight normalized s.c./i.v. dose ratio was 1.04 (0.97-1.11). Despite no significant overall difference, some patients experienced changes in individual dose efficiency response. Three of 24 patients recorded a greater than 30% change, four of 24 recorded between a 20 and 30% change, and five of 24 patients recorded between a 10 and 20% change relative to i.v. dose efficiency. CONCLUSIONS: This study further supports s.c. and i.v. dosing equality and that overall the more convenient i.v. route can be used with equal dosing efficiency. However, patients switching routes of administration should be monitored due to the wide range in individual response. PMID- 15877672 TI - Prediction of failure of native arteriovenous fistulas using 'on-line' fistula flow measurements. AB - BACKGROUND: Measurement of blood flow within native arteriovenous fistula during haemodialysis is recommended to detect incipient fistula failure. In the present study the value of such flow measurements was assessed in a group of patients on maintenance haemodialysis, with access via native arteriovenous fistulas. METHODS: Flow was measured using the 'on-line' thermodilution technique, on three separate occasions, and correlated with subsequent fistula failure within 6 months. RESULTS: Of the 53 patients studied, there were six failures (three thromboses and three inadequate dialysis filtration rates). Flow rates in patients who progressed to fistula failure were significantly less than flow rates in patients whose fistulas did not fail (U = 13.0, P < 0.0003). Failure was no more common in one type of fistula than another (type fistula: F = 0.29, P = 0.88; flow predicting failure: F = 7.22, P = 0.010). Receiver operating characteristic (ROC) curve analyses confirmed flow measurement to be a useful predictor of fistula failure (area under ROC curve 0.91). The optimal threshold of 576 mL/min flow gave a sensitivity of 89% and a specificity of 81%. Measurement of access resistance was less useful in predicting failure (area under ROC curve 0.87). Measurement of fall in flow from the previous measurement was of no use (area under ROC curve 0.535). CONCLUSION: On-line thermodilution measurement of flow within established native arteriovenous fistula is useful in surveillance and early prediction of fistula failure. Fistula flow <576 mL/min may indicate incipient native fistula failure, and should prompt further investigation. PMID- 15877673 TI - Efficacy of a non-vancomycin-based peritoneal dialysis peritonitis protocol. AB - BACKGROUND: Peritonitis has a significant impact upon morbidity and mortality of peritoneal dialysis (PD) patients. Gram-positive organisms account for the majority of infections and vancomycin is a cost effective broad-spectrum antimicrobial treatment for PD peritonitis, but this may lead to the emergence of multiple antibiotic-resistant organisms. The purpose of the present paper was to evaluate the efficacy of a non-vancomycin-based protocol comprising cephazolin and gentamicin, which was introduced in the present PD population as empirical treatment for peritonitis. METHODS: The study involved 82 peritonitis episodes over a 4-year period in 58 patients, excluding those with previous methicillin resistant staphylococcal peritonitis. RESULTS: With cephazolin and gentamicin there was no apparent difference in response or relapse rates in comparison to reported studies using vancomycin-based first-line therapy protocols. CONCLUSION: We advocate initial treatment of PD peritonitis with non-vancomycin-based therapy given similar efficacy and the potential for reduction of resistant organisms. PMID- 15877674 TI - Erythropoietin-resistant anaemia in a predialysis patient with Klinefelter syndrome. AB - A diabetic predialysis patient who had significantly reduced sensitivity to erythropoietin therapy was admitted to Tsukuba University Hospital. Many factors that might have been the cause of the erythropoietin resistance were examined, and a diagnosis of refractory anaemia was made based on a bone marrow aspiration biopsy. A cytogenetic abnormality (47, XXY) was also detected in the bone marrow biopsy specimen, and hence the patient was also diagnosed with Klinefelter syndrome. It was suspected that the sex chromosome abnormality influenced glucose intolerance, renal insufficiency, and erythropoietin resistance due to myelodysplastic changes in the bone marrow. PMID- 15877675 TI - Direct comparison of repeated same-day self and ambulatory blood pressure monitoring. AB - BACKGROUND AND AIM: The relative roles of self and ambulatory blood pressure monitoring (self BPM, ABPM, respectively) have yet to be clearly defined despite accumulating evidence of self BPM benefits, particularly in hypertension management. In particular, measurements comparable to ABPM have been documented with self-measurement, usually twice daily over one or more days. Nevertheless, ABPM offers a series of recordings performed throughout the day that can be invaluable during drug treatment. Consequently, this study evaluated 18 normotensive and 79 hypertensive patients, including 21 not taking hypotensive agents, and measured their blood pressure at least every 2 h while they were also undergoing ABPM. METHODS AND RESULTS: Self-measurement was performed between five and 12 times during the day with 77% of subjects preferring self BPM rather than daytime ABPM. When compared with the closest ABP measurement, mean systolic and diastolic pressures were not significantly different throughout the day in both normotensive and hypertensive patients. However, a comparison of a series of ABPM hourly averages (three readings performed closest to the respective self BPM reading) throughout the day in all hypertensive subjects found ABPM readings to be slightly but significantly lower (3.3 systolic, 2.2 mmHg diastolic; P < 0.05). CONCLUSION: It is concluded that multiple self-measurements throughout the day provides comparable information to daytime ABPM, is preferred by most patients, and should be a cost-effective alternative in the diagnosis and management of hypertension. PMID- 15877676 TI - New insights into the molecular biology of the glomerular filtration barrier and associated disease. AB - The glomerular filtration barrier of the kidney can no longer be considered as an inert and adynamic structure, viewed by electron microscopy. Molecular biology, medical genetics and protein chemistry have enabled us to further understand the complex structure and function of this highly specialized barrier of the kidney. Minor aberrations of physiology can lead to fatal disease. Recent advances in the understanding of the physiology of endothelial cells, glomerular epithelial cells and the glomerular basement membrane and its components, and how these relate to disease, will be considered systematically. PMID- 15877677 TI - Heparanase inhibition reduces proteinuria in a model of accelerated anti glomerular basement membrane antibody disease. AB - BACKGROUND: The beta-d-endoglycosidase, heparanase, is emerging as an important contributor to the pathogenesis of proteinuria. The purpose of the present study therefore was to examine the role of heparanase in a model of accelerated anti glomerular basement disease (anti-GBM). METHODS: Accelerated anti-GBM disease was induced and animals sacrificed at day 10 to establish heparanase expression using immunohistochemistry and western blot analysis. In addition, cortex was isolated from normal and diseased glomeruli to determine if mRNA levels altered with disease. A previously validated anti-heparanase antibody associated with proteinuria reduction, in a model of membranous nephropathy, was administered prior to disease induction to establish its impact on protein excretion in this model. RESULTS: At day 10 of anti-GBM disease, an increase in glomerular heparanase was shown using immunohistochemistry. Sequential staining studies revealed that this increase was associated with glomerular endothelial, epithelial cells and invading ED-1-positive inflammatory cells. RT-PCR revealed an insignificant 1.2-fold induction of mRNA at day 10 of disease. Western blot analysis of kidney cortex confirmed that the active 58-kDa heparanase species was restricted to diseased kidney at day 10. The inactive 65-kDa precursor, however, was found only in cortex derived from normal kidney. Proteinuria at day 10 of disease was significantly reduced, in the absence of altered rat anti-sheep antibody titres, after administration of a validated polyclonal anti-heparanase antibody (P < 0.05). Furthermore, sheep IgG deposition was not altered by administration of the anti-heparanase antibody. CONCLUSION: These data suggest that heparanase contributes to the pathogenesis of proteinuria in a model of anti GBM disease. PMID- 15877678 TI - Pathogenesis of lupus nephritis: an update. AB - Lupus nephritis (LN) is a prototypic autoimmune disease. Its immunopathogenesis is characterized by the loss of self-tolerance. In this article, we review our current understanding of the disease mediators of LN. There is ample evidence to suggest a pathogenic role of nephritogenic autoantibodies. These antibodies cross react with nucleosomal epitopes, and the in vivo generation of nucleosomes requires apoptosis. Furthermore, there is an intriguing and paradoxical relationship between complement and systemic lupus erythematosus (SLE). Immune complex-mediated activation of complement through the classic pathway is traditionally believed to be a major mechanism by which tissue injury occurs. In contrast, hereditary deficiencies of complement components increase the risk of SLE. Finally, the roles of reactive nitrogen and oxygen species are emphasized. PMID- 15877679 TI - Treatment for lupus nephritis: a revisit. AB - The optimal treatment of severe lupus nephritis remains unclear. Regimens consisting of steroid and cyclophosphamide (CYC) appear to be most effective. Infection and gonadal toxicity is a major concern of CYC use in patients of reproductive age. In addition, failure to respond or refractory to CYC treatment may lead to the development of end-stage renal disease. Mycophenolate mofetil (MMF) is a new immunosuppressive agent that selectively inhibits activated lymphocytes and renal mesangial cells. Data from experimental lupus nephritis and controlled studies, albeit small and lacking statistical power, have revealed that MMF is as effective in lupus patients as CYC in the induction of renal remission or as maintenance therapy to reduce renal flare in the short term. The significantly less ovarian toxicity and infection when compared to CYC are particularly attractive for the consideration of MMF in lupus nephritis. The potential of other new therapeutic agents is discussed together with the need for patient recruitment in future trials of lupus nephritis to address the importance of ethnicity as well as histological grading. PMID- 15877680 TI - Renal hypoplasia and situs inversus totalis. AB - A spectrum of renal abnormalities of patients with situs inversus has been reported. Renal dysplasia is the most common. Herein is described for the first time, an association of situs inversus totalis, unilateral congenital renal hypoplasia and external ear cartilage deformity. PMID- 15877681 TI - Higher rate and earlier peritonitis in Aboriginal patients compared to non Aboriginal patients with end-stage renal failure maintained on peritoneal dialysis in Australia: analysis of ANZDATA. AB - BACKGROUND: Aboriginal patients maintained on peritoneal dialysis (PD) have a higher rate of technique failure than any other racial group in Australia. Peritonitis accounts for the bulk of these technique failures, but it is uncertain whether the increased risk of peritonitis in Aboriginal patients was independent of associated comorbid conditions, such as diabetes mellitus. METHODS: Using data collected by the Australia and New Zealand Dialysis and Transplant Registry (ANZDATA), peritonitis rates and time to first peritonitis were compared between Aboriginal (n = 238) and non-Aboriginal patients (n = 2924) commencing PD in Australia between 1 April 1999 and 31 March 2003. RESULTS: Aboriginal PD patients were younger, and had a higher incidence of diabetes than their non-Aboriginal counterparts. Mean peritonitis rates were significantly higher among Aboriginal (1.15 episodes/year; 95% confidence interval (CI): 1.03 1.28) than non-Aboriginal patients (0.60 episodes/year; 95% CI: 0.57-0.62, P < 0.05). Using multivariate negative binomial regression, independent predictors of higher peritonitis rates include Aboriginal racial origin (adjusted odds ratio 1.78; 95% CI: 1.45-2.19), obesity, age and absence of a recorded dialysate : plasma creatinine ratio (D/P creatinine) measurement. Aboriginal racial origin was also associated with a shorter median time to first peritonitis (9.9 vs 19.3 months, P < 0.05), which remained statistically significant in a multivariate Cox proportional hazards model (adjusted hazard ratio 1.76; 95% CI: 1.47-2.11, P < 0.05). CONCLUSION: Aboriginal and obese PD patients have a higher rate of peritonitis and a shorter time to first peritonitis, independent of demographic and comorbid factors. Further investigation of the causes of increased peritonitis risk in Aboriginal patients is needed. PMID- 15877682 TI - Polyomavirus nephropathy after renal transplantation: a single centre experience. AB - BACKGROUND: Polyomavirus associated nephropathy (PVN) in renal transplant recipients has been observed with increasing frequency recently and has emerged as a cause of allograft failure linked to highly potent new immunosuppressive regimens containing tacrolimus or mycophenolate mofetil (MMF). METHODS: Polyomavirus associated nephropathy was identified in nine out of 182 patients who received renal transplantation between October 1998 and July 2003. PVN was confirmed by allograft biopsy. The clinical records of these nine patients were reviewed, as were all of the allograft biopsies. Electron microscopy was performed in all nine cases. After the diagnosis of PVN, maintenance immunosuppression was reduced. The clinical course and outcome of the PVN patients were reviewed in relation to manipulation of immunosuppressive agents. RESULTS: There were nine cases of PVN in renal transplant recipients and the incidence of PVN was 4.9%. All patients with PVN were under triple immunosuppression comprising tacrolimus and MMF. The mean time to a diagnosis of PVN was 7.8 months after transplantation. Three of the nine patients received antirejection therapy prior to PVN. Seven out of nine PVN patients presenting acute allograft dysfunction were initially treated with high-dose intravenous steroid pulse or OKT3 before reduction of the immunosuppression. After reduction of the immunosuppression, seven patients stabilized their renal function. Two (22%) lost their grafts due to persistent PVN and chronic rejection. Two (22%) patients later developed acute rejection after reduction of the immunosuppression. CONCLUSION: PVN can cause allograft dysfunction and graft loss. Renal allograft recipients who are at risk of PVN should be routinely screened with urine cytology and quantitative measurements of viral load in the blood, particularly patients who had graft dysfunction. Early diagnosis and judicious alteration of immunosuppressive agents might permit a superior prognosis and reduce the graft loss from PVN in renal transplant recipients. PMID- 15877683 TI - Should the AACN, proposed Doctor of Nursing Practice degree, be used as a model for international nursing? PMID- 15877684 TI - Caring for a person with dementia: exploring relationships between perceived burden, depression, coping and well-being. AB - This study was concerned with identifying the impact of variables such as gender, length of time caring, coping style, depression and perception of caregiving burden on the physical and psychological well-being of carers of persons with dementia. Forty-two carers aged between 21 and 88 years from Blue Care's Homecare Dementia Service and Cairns Aged Care Health Service participated in the study. A cross-sectional survey research design was used, with participants providing information on the Satisfaction with Life Scale, the Center for Epidemiologic Studies Depression Scale, the COPE, Short Form (SF)-12 and the Zarit Caregiver Burden Scale. Perceived burden accounted for 41.7% of the variance in satisfaction with life as a subjective measure of well-being. There were no significant differences between male and female carers. Satisfaction with life was not found to decrease with length of time caring for the dementia sufferer. There were no significant findings in regard to coping style or physical health of carers. The well-being of carers can be enhanced through strategies which lead to a reduced perception of burden, with respite services providing tangible relief from burden. PMID- 15877685 TI - Home health nursing care agenda based on health policy in Korea. AB - Home health nursing care (HHNC) in Korea has taken on an important role under the mandate of the national health care system since 2000. This program was developed to verify the possibility of early discharge of hospitalized patients and cost containment through a research and development project that was conducted with the government from 1994 to 1999. The process of development of HHNC provided an opportunity to realize the advancement and changes in the system into a consumer focused structure. This is an important turning point for the Korean health care system that suggests certain possibilities for building a foundation for further changes in the service delivery structure. The structure, which had been limited to a supplier-oriented model, is moving to a consumer-oriented structure. Accordingly, the major function and role of nursing policy makers in Korea is to develop an agenda and alternatives for policy-making in a systematic manner and to present implementation strategies clearly. PMID- 15877686 TI - Hepatitis C prevention with nurses. AB - A state campaign to educate nurses about hepatitis C and encourage nurses to fight the epidemic provided an opportunity to assess the relationship between information and safety practices. Baseline and follow-up surveys were sent to random, representative samples (n = 6000) of licensed nurses in Massachusetts, USA. Responses received by 42% (n = 2443) at baseline and 37% (n = 2167) at follow-up were compared to explore the relationship between receiving information and propensity to counsel patients or use safety equipment. Only data from nurses in direct patient care were analyzed. Although few changes were observed, the assessment provided further evidence that more than a single intervention is needed to achieve health behavior change. The number of nurses who never used safety equipment did decrease slightly, from 13.3% (191) to 8.2% (132), and nurses who received information were more likely to talk to patients about the virus: 49% (414) compared to 21% (120). Hepatitis information aimed at nurses may help control this emerging epidemic. PMID- 15877687 TI - Gender differences in health habits and in motivation for a healthy lifestyle among Swedish university students. AB - The aim of the present study was to investigate gender differences in students' health habits and motivation for a healthy lifestyle. The sample of students comprised a probability systematic stratified sample from each department at a small university in the south-west of Sweden (n = 479). A questionnaire created for this study was used for data collection. Self-rated health was measured by number of health complaints, where good health was defined as having less than three health complaints during the last month. A healthy lifestyle index was computed on habits related to smoking, alcohol consumption, food habits, physical activity and stress. Female students had healthier habits related to alcohol consumption and nutrition but were more stressed. Male students showed a high level of overweight and obesity and were less interested in nutrition advice and health enhancing activities. The gender differences are discussed in relation to the impact of stress on female students' health, and the risk for male students in having unhealthy nutritional habits in combination with being physically inactive and drinking too much alcohol. PMID- 15877688 TI - Essentials of research ethics for healthcare professionals. AB - Whether conducted in a university or the healthcare field, research with human subjects gives rise to a multitude of ethical questions for healthcare professionals. While engaging in ongoing professional development on how to conduct research ethically, both clinicians and scientists need to expand their knowledge to provide answers to the following questions: Which ethical theories serve as a foundation for ethical principles in research ethics? What ethical principles should a researcher respect when conducting research with human subjects? What does it mean to conduct research ethically? What ethical dilemmas are encountered by a researcher in conducting research? This paper provides a review of ethical theories, and the ethical basis of guidelines developed and used to guide human subject research. Ethical behaviors and the personal responsibility of the researcher conducting research with human subjects are discussed along with the ethical considerations in research designs and methods. PMID- 15877689 TI - Nursing time allocation and other considerations for staffing. AB - The purpose of this project was to evaluate how nursing staff on the telemetry unit at Merle West Medical Center, a rural Pacific northwest community medical center in the USA, spend their time on selected nursing tasks, and how they believed staffing and patient acuity levels could best be determined. A self report survey tool was developed, piloted and administered. Of the tasks that were listed in the survey, registered nurses (RNs) reported spending 39% of their time performing tasks that RNs only can perform. RNs reported spending 12% of their time performing activities that certified nursing assistants (CNAs) could perform alone and 49% of their time was spent on tasks that both RNs and CNAs must perform. Results from the quantitative portion of the survey indicated that RNs may not be used efficiently due to task overlap. Administering a modified survey which includes a balance of direct and indirect patient care tasks, as well as only those tasks that are truly performed frequently, will increase the relevance of the results. PMID- 15877690 TI - Childhood obesity in Taiwan: review of the Taiwanese literature. AB - In a number of countries, including Taiwan, the prevalence of childhood obesity has been steadily increasing. A study to assess school nurses' perspective on their role in supporting children and preventing childhood obesity in Taiwan is currently being undertaken. A search of the literature reveals that most research publications come from the West and these studies have been useful. However, it is important to isolate the research and policy materials that take into account the contextually and culturally relevant factors in Taiwan and neighboring countries. Findings from a review of the Taiwanese literature are presented in this paper. The literature reveals the factors associated with the prevalence of childhood obesity and prevention strategies. A significant proportion of the research is medical and focuses on cardiovascular disease rather than health promotion and education. However, there are findings in this review that generally support health promotion activities and programs that are school based. There appears to be an urgent need for investment in research that assesses the long-term effectiveness of interventions designed to promote the maintenance of healthy weight during childhood in the Taiwanese society. Western literature is referred to occasionally in this paper in order to introduce an issue or to compare with a Taiwanese paper. PMID- 15877705 TI - Anxiety disorders in the elderly: outdated beliefs and a research agenda. PMID- 15877706 TI - A review of studies describing the use of acetyl cholinesterase inhibitors in Parkinson's disease dementia. AB - OBJECTIVE: To review the literature relating to the use of acetyl cholinesterase inhibitors in Parkinson's disease dementia (PDD). METHOD: MEDLINE (1966--December 2004), PsychINFO (1972--December 2004), EMBASE (1980--December 2004), CINHAL (1982--December 2004), and the Cochrane Collaboration were searched in December 2004. RESULTS: Three controlled trials and seven open studies were identified. Efficacy was assessed in three key domains: cognitive, neuropsychiatric and parkinsonian symptoms. CONCLUSION: Cholinesterase inhibitors have a moderate effect against cognitive symptoms. There is no clear evidence of a noticeable clinical effect against neuropsychiatric symptoms. Tolerability including exacerbation of motor symptoms--in particular tremor--may limit the utility of cholinesterase inhibitors. PMID- 15877707 TI - Handedness in schizophrenia: a quantitative review of evidence. AB - OBJECTIVE: The prevalence of various anomalous handedness subtypes in schizophrenia patients remains ambiguous. Although current literature favours the notion that the shift in lateral preferences seen is because of an increase of mixed-handedness, several studies suggest that exclusive left handedness is more prevalent than in the general population. METHOD: Over 40 studies with reported prevalence data on various handedness subtypes in a schizophrenia population were evaluated by meta-analysis. Combined odds ratios for the three common handedness subtypes (left, mixed, and right) were separately calculated. RESULTS: Each of the three atypical hand dominance patterns were significantly greater in schizophrenia patients than in control subjects, showing that the leftward shift in handedness distribution is not entirely because of an increase in mixed handedness alone. CONCLUSION: An increase of exclusive left-handedness is at variance with the prevailing assertion that the handedness shift in schizophrenia patients is because of a diffuse and bilateral hemispheric insult. PMID- 15877708 TI - The outcome of anxiety disorders in older people at 6-year follow-up: results from the Longitudinal Aging Study Amsterdam. AB - OBJECTIVE: To examine long-term outcome of late-life anxiety disorders and utilization of mental health care services. METHOD: A cohort of subjects (aged > or = 55 years) with an anxiety disorder (n = 112) was identified in the Longitudinal Aging Study Amsterdam (n = 3107). At 6 year follow-up, the rate of persistence and prognostic factors for persistence of anxiety were established. RESULTS: Six years after baseline 23% of our sample met the criteria for an anxiety disorder. Another 47% suffered from subclinical anxiety symptoms. Persistence of anxiety was associated with a high score on neuroticism at baseline. Use of benzodiazepines was high (43%), while use of mental health care facilities (14%) and anti-depressants (7%) remained low in those with persistent anxiety. CONCLUSION: Results indicate that those high in neuroticism are at greater risk for persistence of anxiety. Efforts to enhance appropriate referral of anxious older adults do not seem to have had the desired effect. PMID- 15877709 TI - Sertraline in generalized anxiety disorder: efficacy in treating the psychic and somatic anxiety factors. AB - OBJECTIVE: The objective was to study the efficacy of sertraline on symptoms of psychic and somatic anxiety in patients suffering from moderate-to-severe generalized anxiety disorder (GAD). METHOD: Out-patients with DSM-IV GAD were randomized to 12 weeks of double-blind treatment with placebo. The psychic and somatic anxiety factors of the Hamilton Anxiety Rating Scale (HAM-A) and the Quality of Life, Enjoyment, and Satisfaction Questionnaire were analyzed. RESULTS: Treatment with sertraline resulted in significantly greater last observation carried forward (LOCF)-endpoint improvement than placebo on both the HAM-A psychic and somatic anxiety factors. At LOCF-endpoint, all items on the HAM A psychic factor were more improved on sertraline than placebo, as were three of seven items on the somatic factor. Reduction of secondary depressive symptoms was more correlated with endpoint improvement in quality of life than either psychic- or somatic anxiety. CONCLUSION: Sertraline treatment demonstrated efficacy for both the psychic and somatic anxiety symptoms of GAD. PMID- 15877710 TI - Comorbidity of physical and mental disorders and the effect on work-loss days. AB - OBJECTIVE: To examine the association between physical and mental disorders and the separate and joint effect of physical and mental disorders on work-loss. METHOD: Data was derived from the Netherlands Mental Health Survey and Incidence Study. This was a general population study in which 7076 adults, aged between 18 and 64 years, were assessed using the Composite International Diagnostic Interview. Medically treated physical disorders and work-loss were assessed using self-reports. RESULTS: All physical disorders, except injury caused by accident, were significantly related to anxiety and mood disorders, but only weakly related to substance use disorders. Both physical and mental disorders were significantly related to work-loss; mental disorders more so than physical disorders. Physical mental (PM) comorbidity leads to a mainly additive increase in work-loss. CONCLUSION: PM comorbidity is very common in the general population and leads to a greater absenteeism from work than pure disorders that also cause personal and social problems. PMID- 15877711 TI - Does it make sense to do repeated surveys?--the Lundby Study, 1947-1997. AB - OBJECTIVE: To describe the Lundby Study and the difficulties in doing repeated surveys. METHOD: Best-estimate consensus diagnoses have been used since 1957 together with DSM-IV and ICD-10 in 1997. RESULTS: The Lundby population consisting of 3563 probands was investigated in 1947, 1957 and 1972. Sufficient information was available for 98-99%. In 1997-2000 a fourth field investigation was carried out. Attrition rate for the interviews was 13% (238/1797). About 36% (1030/2827) had died between 1972 and 1997, but data from registers, case notes and key-informants for the period 1972 and 1997 completed the information for 94% (2659/2827). The population has followed the same pattern of development as many rural populations in Sweden since the 1940s. Multiple sources of information are preferable in longitudinal studies in order to tackle the problem of changing diagnostic systems. CONCLUSION: Low attrition rates over 50 years and reasonable diagnostic uniformity make comparisons over time justifiable. PMID- 15877712 TI - Adjunctive bright light in non-seasonal major depression: results from patient reported symptom and well-being scales. AB - OBJECTIVE: In this study, we tested the efficacy of bright light therapy as an adjunct to antidepressant treatment (sertraline) in patients with non-seasonal major depression. METHOD: In a randomized double-blind controlled trial, 102 patients were treated for 5 weeks with either white bright light (10.000 lx, 1 h/day) or red dim light (50 lx, 30 min/day). All patients received sertraline in a dosage of 50 mg daily. The self-assessment scales used were the Major Depression Inventory (MDI), the Psychological General Well-Being Scale (PGWB) and the Symptom Check List (SCL-90R). RESULTS: On all three questionnaires the score differences between baseline and endpoint were greatest in the bright light group. On the SCL-90R, the difference reached statistical significance. Results and effect sizes are compared with results from Danish national population studies applying PGWB and SCL-90R. CONCLUSION: The results advocate the use of bright light as an adjunct treatment of non-seasonal depression. PMID- 15877713 TI - Parent ratings of school behaviour in children at risk of attention deficit/hyperactivity disorder. AB - OBJECTIVE: To investigate whether parents are accurate informants of child hyperactivity symptoms and impairment at school. METHOD: Parents of a community sample of 93 children with pervasive hyperactivity completed rating scales about their child's behaviour at home and school. These were compared with teacher ratings. RESULTS: Parent ratings about school correlate more closely with parent (home) than teacher ratings. Such ratings systematically under-estimate teacher ratings and are influenced by the child's behaviour at both home and school as well as parental mental health. However, a parental report of impairment for the child at school is likely to be accurate. CONCLUSION: There are limitations in relying on parental accounts of school behaviour if teacher ratings are unavailable. As such ratings may under-identify children with ADHD and discrepancies between parent and teacher ratings may reflect actual differences in behaviour, this suggests that ratings are required from both sets of informants. PMID- 15877717 TI - Neuroleptic dysphoria 50 years later. PMID- 15877718 TI - Neuroleptic dysphoria: revisiting the concept 50 years later. AB - OBJECTIVE: To review the concept of neuroleptic dysphoria, its historical development and the current state of the art. METHOD: This paper is based on extensive but selective literature review and also draws on our extensive clinical and research experiences. RESULTS: Although the construct of neuroleptic dysphoria was recognized shortly following the introduction of the first antipsychotic, chlorpromazine, it took several years for the concept to receive adequate research and clinical attention. Without having direct evidence to link neuroleptic dysphoria to dopamine, it was generally understood that dopamine played a significant role in its genesis. In recent neuroimaging studies and dopamine depletion strategies, the role of dopamine in the genesis of neuroleptic dysphoria has been directly confirmed. CONCLUSION: Neuroleptic dysphoria is a valid construct, which has significant implications for treatment and outcome. It is now clear that it relates to dopamine activities in the nigrostriatal complex. Recent research has also raised the issue of whether neuroleptic dysphoria is a variant of extrapyramidal symptoms. Meanwhile, the role of dopamine in both the genesis of neuroleptic dysphoria and addictive behaviour has raised the issue of both conditions being different facets of the schizophrenic disease process. The recent interface of addiction and psychiatry research may have opened a new science: the science of subjective tolerability disorders. PMID- 15877719 TI - The role of dopamine in reward and pleasure behaviour--review of data from preclinical research. AB - OBJECTIVE: The purpose of this article is to review some of the basic aspects of the dopaminergic system and its role in reward and pleasure behaviour. We also discuss the association between dopamine and unpleasant symptoms that are commonly found in neuropsychiatric disorders and may also be side-effects of neuroleptic drugs. METHOD: A computer-based search of the literature, augmented by extensive bibliography-guided article reviews, were used to find basic information on the dopamine and the reward systems, and symptoms such as dysphoria, anhedonia and depression. RESULTS: Central dopaminergic neurotransmission is complex, having multiple actions at each level of the mesocorticolimbic reward pathway. The role of dopamine in the reward process was classically associated with the ability to experience pleasure; recent data suggest a more motivational role. Dysfunction of the dopamine transmission in the reward circuit is associated with symptoms such as anhedonia, apathy and dysphoria found in several neuropsychiatric disorders, including Parkinson's disease, depression, drug addiction, and neuroleptic-induced dysphoria. CONCLUSION: Viewing the dysfunctions of the reward pathways within a broader spectrum and exploring its complex relations with the dopaminergic transmission may help understand the pathophysiology of these neuropsychiatric disorders and lead to a rational development of novel treatments. PMID- 15877720 TI - Adolescence, schizophrenia and drug abuse: a window of vulnerability. AB - OBJECTIVE: To discuss the neurobiological and psychosocial developmental factors in adolescence contributing to simultaneous onset and co-occurrence of psychosis and substance use disorders. METHOD: A review of the literature. RESULTS: Adolescence is a period with specific psychosocial challenges and specific changes in the brain that increase the probability of the onset of both psychosis and substance abuse, in predisposed people. In vulnerable adolescents it is proposed that an excessive pruning of dopaminergic neurones leads to mesocortical hypofrontality causing anhedonia and dysphoria. At the same time, anhedonia and dysphoria are important risk factors for the development of substance abuse. In turn, hypofrontality leads to a reduction in mesocortical feedback inhibition of the mesolimbic system resulting in aberrant salience and positive symptoms. Finally, the development of aberrant salience plays a role in both psychoses and craving. CONCLUSION: Attention should be paid to the interaction of drug abuse and schizophrenia and an integrated treatment is needed. Dysphoria and anhedonia in schizophrenic adolescents are important factors in treatment with antipsychotic medication, both in terms of patient satisfaction and in the prevention of substance abuse. PMID- 15877721 TI - Subjective experiences on antipsychotic medications: synthesis and conclusions. AB - OBJECTIVE: This report synthesizes the literature describing the phenomenology, clinical importance and biology of subjective responses to antipsychotic medications in schizophrenia. A patient's experience of an antipsychotic is important because unpleasant or dysphoric responses can impair therapeutic relationships, lead to medication non-adherence, and have direct negative effects on a patient's quality of life. METHOD: The author selectively reviewed early studies of subjective responses to antipsychotics and integrated this literature with the work of the other investigators in this special section. RESULTS: There is substantial evidence that second-generation antipsychotics have advantages in causing fewer dysphoric responses when compared with first-generation agents. Clinical and neuroimaging studies suggest that dopamine blockade is an important determinant of many of these dysphorias. At this point in time it is unclear whether dysphoria results from extrapyramidal symptoms--particularly akathisia and akinesia--or whether they are a direct result of decreased dopamine activity. CONCLUSION: Clinicians and researchers should continue to monitor dysphorias in schizophrenia. Contributions by the authors in this supplement provide new and more refined methods for measuring subjective responses in future studies. PMID- 15877722 TI - Technical considerations when obtaining and interpreting prostatic biopsies from men with suspicion of early prostate cancer: part I. PMID- 15877723 TI - Technical considerations when obtaining and interpreting prostatic biopsies from men with suspicion of early prostate cancer: part 2. PMID- 15877724 TI - Gleason grading of prostate cancer in needle biopsies or radical prostatectomy specimens: contemporary approach, current clinical significance and sources of pathology discrepancies. AB - The Gleason grading system is a powerful tool to prognosticate and aid in the treatment of men with prostate cancer. The needle biopsy Gleason score correlates with virtually all other pathological variables, including tumour volume and margin status in radical prostatectomy specimens, serum prostate-specific antigen levels and many molecular markers. The Gleason score assigned to the tumour at radical prostatectomy is the most powerful predictor of progression after radical prostatectomy. However, there are significant deficiencies in the practice of this grading system. Not only are there problems among practising pathologists but also a relative lack of interobserver reproducibility among experts. PMID- 15877725 TI - Therapeutic approaches in metastatic renal cell carcinoma. PMID- 15877726 TI - Severe conidiobolomycosis complicating induction chemotherapy in a patient with acute lymphoblastic leukaemia. PMID- 15877727 TI - Loss of skin pigment caused by imatinib therapy. PMID- 15877728 TI - Chronic sickle cell lung disease: new insights into the diagnosis, pathogenesis and treatment of pulmonary hypertension. AB - Pulmonary hypertension is a common complication of sickle cell disease (SCD). In spite of the mild elevations in pulmonary artery pressures in these patients, the associated morbidity and mortality is high. In fact, in adult patients with SCD, pulmonary hypertension is emerging as the major independent risk factor for death. The aetiology of pulmonary hypertension is probably multifactorial, including haemolysis, impaired nitric oxide bioavailability, chronic hypoxaemia, thromboembolism, parenchymal and vascular injury because of sequestration of sickle erythrocytes, chronic liver disease and asplenia. Interestingly, pulmonary hypertension is emerging as a common, and probably, invariant sequella of lifelong haemolytic anaemia in other hereditary and acquired haemolytic diseases, such as thalassaemia, stomatocytosis and spherocytosis. There are currently limited specific data on the effects of any treatment modality for pulmonary hypertension in patients with SCD. It is likely that maximization of SCD therapy, in all patients, and treatment with selective pulmonary vasodilators and antiproliferative agents, in patients with severe disease, would be beneficial. A large trial evaluating the effects of therapy for pulmonary hypertension in the SCD population is clearly indicated. PMID- 15877729 TI - Predicting clinical severity in sickle cell anaemia. AB - The ability to predict the phenotype of an individual with sickle cell anaemia would allow a reliable prognosis and could guide therapeutic decision making. Some risk factors for individual disease complications are known but are insufficiently precise to use for prognostic purposes; predicting the global disease severity is not yet possible. Genetic association studies, which attempt to link gene polymorphisms with selected disease subphenotypes, may eventually provide useful methods of foretelling the likelihood of certain complications and allow better individualized treatment. PMID- 15877730 TI - Bone involvement in sickle cell disease. AB - Bone involvement is the commonest clinical manifestation of sickle cell disease both in the acute setting such as painful vaso-occlusive crises, and as a source of chronic, progressive disability such as avascular necrosis. Management of these problems is often difficult because of the diagnostic imprecision of most laboratory and imaging investigations and because of the lack of evidence for most surgical procedures in sickle cell disease. This review first discusses the acute problems related to bone involvement in sickle cell disease, with particular reference to differentiating infection from infarction, and then describes the long-term effects of sickle cell disease on bone mineral density, growth, and chronic bone and joint damage. PMID- 15877731 TI - Co-existence of multiple subclones in TEL-AML1 at diagnosis of acute lymphoblastic leukaemia in association with submicroscopic deletion of AML1. AB - The TEL/AML1 (ETV6/RUNX1) fusion gene is the most common genetic rearrangement in paediatric acute lymphoblastic leukaemia (ALL). Although considered to be a low risk leukaemia, it is associated with a relapse rate of 10-20%. The coexistence of different subclones at diagnosis, based on polymerase chain reaction (PCR) studies of IG/TCR gene rearrangement, with differential response to chemotherapy, was recently reported in this subtype of ALL. We wished to demonstrate such subclones at diagnosis by a recently developed technique of quantitative multiparametric fluorescence in situ hybridization (FISH). Bone marrow cells from 80 paediatric patients with ALL at diagnosis were analysed for the presence of the TEL/AML1 fusion gene by interphase FISH. Fourteen patients were positive for the translocation. Four of them had several subclones associated with various combinations of additional chromosomal abnormalities. The most striking was an atypical and unexpected hybridization pattern consistent with a submicroscopic deletion of the 5' region of the AML1 breakpoint. Other abnormalities included TEL deletion, trisomy and tetrasomy 21 as well as double TEL-AML1 fusion. The presence of numerous subclones in about 25% of patients with TEL/AML1+ ALL suggests extensive clonal evolution by the time of diagnosis. PMID- 15877732 TI - Immunoglobulin gene segment usage, location and immunogenicity in mutated and unmutated chronic lymphocytic leukaemia. AB - The mutational status of the variable region of the immunoglobulin heavy chain gene (IgV(H)) is an important prognostic marker in B-cell chronic lymphocytic leukaemia (B-CLL), with mutated patients having improved outcome. To examine the impact of mutational status on V(H), D(H), and J(H) gene segment location and immunogenicity, we analysed 375 IgH sequences from 356 patients with B-CLL. Although V(H) and D(H) gene usage was different in mutated compared to unmutated patients, there was no impact of gene location on frequency of use or clinical outcome. Surprisingly, somatic mutations did not increase the immunogenicity of the Ig, as assessed by predicted binding affinity of Ig-derived peptides to major histocompatibility Class I and Class II molecules. Even excluding patients using V(H)1-69, cases using the V(H)1 gene family had a poor outcome. Both mutated and unmutated CLL patients demonstrated evidence of antigen selection. The worst outcome was seen in the subset of 14 unmutated patients with similar HCDR3 amino acid sequence using V(H)1-69, D(H)3-3 and J(H)6, suggesting an antigen-driven process modulating the clinical course. PMID- 15877733 TI - Phenotype and frequency of Epstein-Barr virus-infected cells in pretreatment blood samples from patients with Hodgkin lymphoma. AB - An accumulating body of data suggests that the Epstein-Barr virus (EBV), a lymphotropic herpesvirus, is involved in the pathogenesis of a proportion of cases of Hodgkin lymphoma (HL). In this study, we showed that the frequency of circulating EBV-infected cells was significantly higher (P < 0.001) in pretreatment blood samples from EBV-associated cases when compared with non-EBV associated cases. We further showed that in patients with EBV-associated disease, the virus persisted in the peripheral blood in memory B cells. This phenotype is consistent with that seen in healthy seropositive controls, post-transplant patients and patients with acute infectious mononucleosis. The data suggest that an increased frequency of EBV carrying B cells in peripheral blood is associated with EBV-associated HL. PMID- 15877734 TI - Interphase molecular cytogenetic screening for chromosomal abnormalities of prognostic significance in childhood acute lymphoblastic leukaemia: a UK Cancer Cytogenetics Group Study. AB - Summary Interphase fluorescence in situ hybridization (iFISH) was used independently to reveal chromosomal abnormalities of prognostic importance in a large, consecutive series of children (n = 2367) with acute lymphoblastic leukaemia (ALL). The fusions, TEL/AML1 and BCR/ABL, and rearrangements of the MLL gene occurred at frequencies of 22% (n = 447/2027) (25% in B-lineage ALL), 2% (n = 43/2027) and 2% (n = 47/2016) respectively. There was considerable variation in iFISH signal patterns both between and within patient samples. The TEL/AML1 probe showed the highest incidence of variation (59%, n = 524/884), which included 38 (2%) patients with clustered, multiple copies of AML1. We were thus able to define amplification of AML1 as a new recurrent abnormality in ALL, associated with a poor prognosis. Amplification involving the ABL gene, a rare recurrent abnormality confined to T ALL patients, was identified for the first time. The use of centromeric probes revealed significant hidden high hyperdiploidy of 33% and 59%, respectively, in patients with normal (n = 21/64) or failed (n = 32/54) cytogenetic results. The iFISH contributed significantly to the high success rate of 91% (n = 2114/2323) and the remarkable abnormality detection rate of 89% (n = 1879/2114). This study highlights the importance of iFISH as a complementary tool to cytogenetics in routine screening for significant chromosomal abnormalities in ALL. PMID- 15877735 TI - Expression of the Rho-family GTPase gene RHOF in lymphocyte subsets and malignant lymphomas. AB - We have studied the expression of RHOF, a member of the Rho-GTPase family, in an array of lymphoid cells and tissues. Previous microarray studies demonstrated RHOF upregulation in a subset of transformed follicular lymphomas. Real-time quantitative polymerase chain reaction evaluated RHOF expression in lymphocyte subpopulations, and normal and malignant lymphoid tissue. Cells and tissues of B cell origin expressed higher RHOF levels than their T-cell counterparts. Neoplastic cells and tissues of B-cell origin expressed higher levels of RHOF than their benign cellular counterparts. Relatively elevated levels of RHOF were seen in lymphomas derived from germinal centre origin. PMID- 15877736 TI - Use of mycophenolate mofetil for chronic, refractory immune cytopenias in children with autoimmune lymphoproliferative syndrome. AB - Autoimmune lymphoproliferative syndrome (ALPS) is a disorder of apoptosis associated most often with heritable FAS mutations leading to lymphadenopathy, hypersplenism and chronic refractory autoimmune cytopenias. Mycophenolate mofetil (MMF) was used to treat cytopenias in 13 ALPS patients aged 9 months to 17 years from a cohort of 118 children (aged < 18 years) and 82 adults. Twelve responded for a median follow-up of 49 weeks (range 38-240 weeks), defined by maintenance of adequate blood counts and reduction in dosage or cessation of other immunosuppressive agents. This preliminary experience suggests that MMF may spare steroid usage in patients with ALPS-associated cytopenias. PMID- 15877737 TI - The absence of CD56 on malignant plasma cells in the cerebrospinal fluid is the hallmark of multiple myeloma involving central nervous system. AB - By immunohistochemistry, the CD56-positive myeloma cells were detected in three (38%) bone marrow (BM) and one (13%) cerebrospinal fluid (CSF) samples from eight patients with multiple myeloma involving the central nervous system (CNS MM). Of the three patients with CD56-positive BM myeloma cells, two had CSF myeloma cells negative for CD56. In a control cohort of 84 MM patients without CNS involvement, the BM myeloma cells were CD56-positive in 68 (80%) cases (P < 0.0001). The prevalence of CD56-negative myeloma cells in the BM and CSF of our patients suggests that CD56 downregulation may play a role in the pathogenesis of CNS MM. PMID- 15877738 TI - Chimaerism analyses and subsequent immunological intervention after stem cell transplantation in patients with juvenile myelomonocytic leukaemia. AB - Chimaerism analysis was performed by polymerase chain reaction amplification of short-tandem repeat markers in 30 children following haematopoietic stem cell transplantation for juvenile myelomonocytic leukaemia (JMML). Fourteen patients always had complete chimaerism (CC); one of them relapsed after the discontinuation of the study and 13 continued in complete remission (CR). Mixed chimaerism (MC) was noted in 16 patients. Of those 12 patients demonstrated increasing MC (i-MC); 10 relapsed and two achieved CC following discontinuation of immunosuppressive therapy (IST). Four other patients demonstrating only transient MC are alive in CR. MC with up to 20% of autologous cells could be successfully eradicated without induction of severe graft-versus-host disease when IST was reduced or discontinued directly after the first demonstration of MC. At the same time, MC with up to 10% of autologous cells could disappear without intervention. The interval between MC and relapse ranged from 0-320 d (median 38 d). Donor leucocyte infusion was given to six patients with i-MC, but only one patient responded. Peripheral blood seems as valuable as bone marrow for chimaerism studies. In conclusion, serial quantitative chimaerism studies can identify patients with i-MC who are at high risk for relapse of JMML. Immediate withdrawal of IST is advised in these patients. PMID- 15877739 TI - Brain parenchymal damage after haematopoietic stem cell transplantation for severe sickle cell disease. AB - Prospective magnetic resonance imaging (MRI), magnetic resonance angiography (MRA), neuropsychological testing and neurological examinations were performed to determine the long-term effect of successful haematopoietic stem cell transplantation on the neurological status of nine children with sickle cell disease. A scoring system for severity of brain parenchymal and vascular lesions was developed and applied. Neurological examinations and neuropsychometric tests were stable, but MRI and MRA studies were not. Transient changes occurred early in two patients. Persistent changes occurred in five. Parenchymal lesions occurred in zero of two patients without prior lacunae or infarcts and in all seven with prior lacunae or infarcts (P = 0.0278). PMID- 15877740 TI - Relevance of the criteria commonly used to diagnose myeloproliferative disorder in patients with splanchnic vein thrombosis. AB - Myeloproliferative disorders (MPD) are reported in 25-65% of patients with splanchnic vein thrombosis (SVT). Diagnostic criteria for MPD have not been fully established in this context. Using clusters of abnormal megakaryocytes in bone marrow (BM) biopsy as a reference standard for Philadelphia negative MPD, we assessed the relevance of other criteria currently recommended for the diagnosis of MPD in SVT (128 consecutive SVT patients). First, usual criteria were compared with BM results: endogenous erythroid colony formation (EEC) was strongly correlated with BM results; splenomegaly, blood cell count, total red cell volume, erythropoietin level and cytogenetic were much less accurate. Then, patients were assigned to three groups according to the combination of BM and EEC findings (group I: both present; group II: both absent; group III: other patients); clinical presentation and outcome were compared in each group. At a mean follow-up of 6.09 +/- 6.6 years, progression to a severe form of MPD occurred in 7 of 31 group I patients (23%), in 1 of 34 group III patients (3%) and 0 of 63 group II patients. The combination of marked splenomegaly and platelet count >200 x 10(9)/l was restricted to groups I and III. In conclusion, in patients with SVT, BM findings and EEC allowed the diagnosis of MPD at risk of aggravation. Marked splenomegaly in association with platelet counts >200 x 10(9)/l constitute a simple index with high specificity but low sensitivity. PMID- 15877741 TI - Protein Z-dependent protease inhibitor W303X mutation in venous thrombosis. PMID- 15877742 TI - Bone marrow biopsy in thrombocytopenic or anticoagulated patients. PMID- 15877744 TI - The role of natural killer T (NKT) cells in immune thrombocytopenia: is strong in vitro NKT cell activity related to the development of remission? PMID- 15877745 TI - New formation of periodontal tissues around titanium implants in a novel dentin chamber model. AB - Direct bone-to-implant contact, defined as "osseointegration", is considered most optimal for long-term stability and survival of dental implants. However, the possibility of the formation of a tooth-like attachment apparatus around implants has also been demonstrated. The purpose of this study was to explore the formation of periodontal tissues around titanium implants using a novel and unique experimental model. After resection of the crowns of the maxillary canine teeth in nine mongrel dogs, the roots were hollowed to a depth of 5 mm leaving a thin dentinal wall. Slits were prepared in the cavity wall to create passages from the chamber to the periodontal ligament area. A custom-made, titanium implant was placed into the center of each chamber. Machined, titanium plasma sprayed (TPS) and sand blasted with large grit and acid attacked (SLA) surfaces were used. A collagen barrier was placed over the submerged chamber. Following 4 months of healing, jaw sections were processed for histology. Newly formed periodontal ligament, alveolar bone, and root cementum filled the space between the implant and the wall of the chamber. Ingrown bone was neither in contact with dentin nor with the implant. Thus, an interposed soft connective tissue layer was present. Healing by fibrous encapsulation was observed around most implants. However, cellular cementum was deposited on one TPS and one SLA implant and on the dentinal walls of the chamber. This study shows a remarkable capacity for new periodontal tissue formation at a site where no such tissues ever existed. Maintenance of original periodontal tissue domains most likely prevented osseointegration of the implants. The cementum layer deposited on two implants was likely formed through cementoconductivity rather than by differentiation of periodontal ligament cells upon contact with the implant surface. PMID- 15877746 TI - The impact of nicotine on bone healing and osseointegration. AB - OBJECTIVES: To examine the short-term effect of nicotine on bone healing and osseointegration. MATERIAL AND METHODS: Sixteen female rabbits were divided into two groups. The test group was exposed to nicotine tartrate for 8 weeks and the control group was exposed to placebo. Nicotine or placebo was administered via a miniosmotic pump and plasma cotinine levels were measured weekly. The pump delivered 15 mg of nicotine/day for the animals in the test group. All rabbits had three tibial bone preparations. In the proximal and distal bone bed, implants were placed after 4 weeks (right tibia) and after 6 weeks (left tibia). Thus, 2- and 4-week healing groups were created. Removal torque test (RMT) was performed at the distal implants. Ground sections were made from the proximal and the central bone beds. The fraction of mineralized bone in contact to the implant (BIC) and the bone density within the implant threads (BD-i) were determined for the bone-implant specimens. For the central bone beds without implants the bone density (BD-c) in the bone defects was determined. RESULTS: No significant difference in RMT values was found between the test and the control group. Histomorphometric measurements of the BIC and the peri-implant BD-i showed no significant differences between the test and the control group after 2 or 4 weeks. Significant differences were, however, found between the 2- and 4-week samples. In the central bone beds, there was no significant difference in BD-c between the test and the control group. CONCLUSION: Nicotine exposure in a short period of time did not have a significant impact on bone healing or implant osseointegration in rabbits. PMID- 15877747 TI - Microbiological and clinical outcomes and patient satisfaction for two treatment options in the edentulous lower jaw after 10 years of function. AB - BACKGROUND: Long-term data on microbiological and clinical outcome as well as on patient satisfaction after implant therapy in the edentulous mandible are limited. Especially comparisons between fixed full prostheses (FFPs) and overdentures (ODs), or between anchoring systems for the latter are scarce. AIM: This study aimed to evaluate both of these parameters at the 10-year follow-up in a group of fully edentulous patients rehabilitated via an OD or a FFP (the latter to allow inter-group comparison). MATERIAL AND METHODS: A total of 37 fully edentulous patients (25 ODs, 12 FFPs, age at implant installation ranged from 36 to 85 years) participated in this study. All subjects received their implants (Branemark System, Nobel Biocare AB, Gothenburg, Sweden) 10 years previously. For the ODs different attachment systems (bar, magnets, ball) had been applied that allowed a further intra-group comparison. At the follow-up visit, 10 years after the abutment insertion, a series of periodontal parameters were recorded, long cone radiographs were taken and subgingival plaque samples were collected for analysis using checkerboard DNA-DNA hybridization. The clinical and radiographic data were recorded at abutment connection and after 1 and 10 years. RESULTS: After 10 years of loading, mean plaque and bleeding indices and changes in attachment or marginal bone level were not significantly different, neither between the OD and FFP group, nor within the OD group. The marginal bone loss between abutment connection and year 10 was 0.86 and 0.73 mm for OD and FFP groups, respectively. The subgingival microbiota at implant sites from all (sub) groups was comparable, with low numbers of DNA counts (+/-10 x 10(5)) but high detection frequencies of Actinobacillus actinomycetemcomitans (>90%), Porphyromonas gingivalis (>85%) and Tannerella forsythensis (30%). The composition of the subgingival microbiota was influenced by probing depth and bleeding tendency. Patient satisfaction was very high for both types of prosthetic rehabilitation. The FFP group scored only slightly better for chewing comfort and general satisfaction. CONCLUSION: These data indicate that from the clinical and microbiological standpoint, as well as patient satisfaction, both an OD and a FFP offer a favourable long-term outcome. PMID- 15877748 TI - Treatment of peri-implantitis by the Vector system. AB - AIM: To compare the effectiveness of treatment of peri-implantitis with a novel ultrasonic device, the Vector system, with that of subgingival debridement with carbon fiber curettes. MATERIAL AND METHODS: The study, comprising 11 patients with at least two screw type implants with bleeding on probing (BOP), probing pocket depth (PPD) > or =5 mm, and at least 1.5 mm radiographic bone loss and exposed implant threads, was carried out as a single blind randomized clinical trial. At baseline one randomly chosen implant in each patient was treated by the Vector system (test) while the other implant (control) was treated by submucosal debridement with a carbon fiber curette. After 3 months, the same treatments were repeated. Plaque, BOP, and PPD were recorded on all implant surfaces at baseline, and after 3 and 6 months. Bone levels were recorded on radiographs taken prior to the start of the study, and after 6 months. RESULTS: Oral hygiene around both test and control implants was improved at 3 and 6 months compared with baseline. At 6 months, four of the Vector-treated sites, and only one site treated with curettes, had stopped to bleed. In neither the test nor the control group, were there any differences between baseline and 6 months regarding PPD and bone levels. CONCLUSION: Although there was a greater reduction in the number of sites with BOP following treatment with the Vector system than following instrumentation with carbon fiber curettes, there was no significant difference between the two methods. PMID- 15877749 TI - Long-term results and survival rate of implants treated with guided bone regeneration: a 5-year case series prospective study. AB - One of the key factors for attaining osseointegration is the presence of an adequate osseous volume. In patients with inadequate osseous width or height, a bone augmentation using the guided bone regeneration (GBR) concept may be applied either with a simultaneous or a staged approach. The aim of this multicenter prospective case series study was to evaluate the efficacy and predictability of the GBR technique (simultaneous approach) in patients with peri-implant osseous defects, both dehiscences and fenestrations. Results 5 years post-treatment (survival rates and marginal bone level) were assessed. A total of 19 consecutive patients with 26 peri-implant osseous defects (20 dehiscences and six fenestrations) were treated during the period from September 1992 to June 1993 with a simultaneous GBR approach using non-resorbable membranes combined with autogenous bone grafts or decalcified freeze-dried bone allograft. The mean osseous augmentation was 94.8%. Marginal bone levels at re-entry and 5 years after surgery were calculated from standardized periapical radiographs. One implant was lost 3 months after loading. Thus, the cumulative survival rate was 96.1% after 5 years. The mean marginal bone level after 5 years was 2.03 mm (SD=+/-0.5), without a difference between mesial and distal sites. This study demonstrates that implants with peri-implant defects that are treated with GBR had similar survival rates and crestal bone levels compared with implants in native bone. PMID- 15877750 TI - A prospective 3-year study of fixed bridges linking Astra Tech ST implants to natural teeth. AB - BACKGROUND: Connecting teeth and osseointegrated implants in fixed reconstructions is not generally recommended because of differences in their response to loading. AIM: The aim of the present study was to assess the clinical and radiographic performance of the teeth and implants used to support three unit fixed bridges subjected to normal functional loads. SUBJECTS AND METHODS: Nineteen subjects (10 males, nine females, age range 27-65 years) with an edentulous posterior free end saddle in either maxilla or mandible (Kennedy Class 2), and opposing natural teeth or a tooth-supported fixed bridge were treated and completed the 3-year trial. An Astra Tech ST implant (length: 9 mm (n=2), 11 mm (n=9) or 13 mm (n=8); diameter: 4.5 mm) was placed immediately distal to the last tooth or leaving a single premolar sized space. The distal tooth received a gold coping and the implant was restored with a customised Prepable abutment (Astra Tech Profile BiAbutment: diameter 5.5 or 7 mm). A fixed bridge was placed linking the gold coping and implant abutment either with the pontic as a distal cantilever (n=6, length 7-8 mm) or as a fixed-fixed design (n=13, length 6-12 mm). Standardised radiographs and clinical records were taken at delivery of the prosthesis (baseline BL) and annually. RESULTS: Plaque scores at implant sites increased between BL and subsequent years (P<0.02). Statistically significant increases in probing depth were observed at both abutment teeth and implants between baseline and subsequent years (P<0.001). Marginal bone levels (mm) at the implant and tooth were stable between BL, 1-, 2- and 3-year examinations (implant: BL 0.65+/-0.42, 1 year 0.63+/-0.47, 2 years 0.88+/-0.55, 3 years 0.78+/ 0.64; tooth: BL 2.29+/-0.82, 1 year 2.41+/-0.8, 2 years 2.38+/-1.02, 3 years 2.68+/-0.86). No signs of the intrusion of the abutment teeth were detected. One case of abutment screw loosening occurred. Eight bridges required re-cementation with a permanent cement in place of the temporary cement. There were eight subjects presenting with fractures/chips to the composite component of the bridges. CONCLUSION: The 3-year results demonstrate fully functional successful restorations with no evidence of tooth intrusion and with stable bone levels at both teeth and implants. PMID- 15877751 TI - Oxidized titanium implants (Nobel Biocare TiUnite) compared with turned titanium implants (Nobel Biocare mark III) with respect to implant failure in a group of consecutive patients treated with early functional loading and two-stage protocol. AB - OBJECTIVES: The purpose of this study was to compare two implant types of similar shape but with different surfaces with respect to implant failure. MATERIAL AND METHODS: A total of 136 patients were treated with Nobel Biocare Implants between January 2001 and December 2002. Totally 394 implants were placed, of which 199 were oxidized titanium implants (Nobel Biocare TiUnite and 195 with turned titanium surface (Nobel Biocare Mark III. Sixty-three patients underwent a one stage surgical protocol, of which 24 were objected to early functional loading. The remaining 73 patients were treated with a traditional two-stage surgical protocol. All patients were followed for a minimum of 5 months after loading of the implants. Implants were classified as survivals when clinically stable and fulfilling purported function without any discomfort to the patient, with no signs of infection or ongoing pathologic process. RESULTS: Seven implants were lost in five patients (two males and three females), six in the maxilla and one in the mandible. All failed implants were Mark III implants, inserted following the traditional two-stage protocol. The implant success rate was 98.2% for the whole-patient group, divided as a 100% success rate following the implants with oxidized surface (Nobel Biocare TiUnite compared with a success rate of 96.4% with implants with turned surface (Nobel Biocare Mark III). PMID- 15877752 TI - Zygoma implant-supported midfacial prosthetic rehabilitation: a 4-year follow-up study including assessment of quality of life. AB - OBJECTIVE: Successful prosthetic rehabilitation is crucial for quality of life in cases of large maxillary defects when surgical reconstruction is not advisable because of general health or patient refusal. For this purpose, the extended indications for Zygomaticus fixtures in different defect types were evaluated. PATIENTS AND METHODS: Twelve patients received 28 zygoma implants and 23 dental implants (if a segment of alveolar process was available) and were followed-up 14 53 months. Zygoma implants were positioned classically in the maxillary molar region and to reduce leverage, a premolar and a canine position was developed. The quality of life was assessed by a validated questionnaire after complete rehabilitation. RESULTS: Cumulative zygoma implant survival was 82%. Three losses occurred because of persistent infection and gradual loosening. Lost implants were immediately replaced in adjacent bone. Insufficient implant length within soft tissue reconstructions was prone to chronic infection by pocketing and recurrent overgrowth of granulating tissue. Longer implants were free of soft tissue inhibition, yet prone to overloading and high leverage in cases when no anterior alveolar process and dental implants were present. Zygoma implant success was therefore 71%, including the new premolar and canine Zygomaticus fixture-position. Periotest values increased from 0 to +7 to the fourth year, peri-implant bleeding and plaque index were decreasing from 56% to 0% and 33% to 0%, respectively, and good general quality of life with the priorities on chewing and activity was noted. CONCLUSION: Zygoma implants can reliably anchor the midfacial maxillary prostheses and enable a quality of life comparable with autologous maxillary reconstruction. They can be replaced immediately if local infection or loosening should occur. A premolar and canine position reduce leverage when no anterior alveolar process is present. The patient can alternatively be provided with dental implants. PMID- 15877753 TI - Technical and biological complications/failures with single crowns and fixed partial dentures on implants: a 10-year prospective cohort study. AB - OBJECTIVES: To assess prospectively over 10 years the incidences of technical and/or biological complications and failures occurring in a cohort of consecutive partially edentulous patients with fixed reconstructions on implants of the ITI Dental Implant System. METHODS: Eighty-nine patients were available, 34 (38.2%) were male, 55 (61.8%) were female. At the 10-year examination (range 8-12 years), they were 58.9 years old (range 28-88 years). RESULTS: Single crowns (SC): 48 patients had been restored with 69 SC on 69 implants. Five of the implants with the crowns were lost because of biological failures. Two crowns (2.9%) were remade because of technical failures. Total failure amounted to seven (10%). Implant borne fixed partial dentures (I-I FPD): In 29 patients who had been restored with 33 implant borne suprastructures, the total number of failed I-I FPD was 2 (6.1%). Tooth-implant borne fixed partial dentures (I-T FPD): In 21 patients, 22 mixed tooth-implant borne reconstructions were constructed. The number of failed FPD reached 7 (31.8%). Statistically significantly fewer biological failures occurred with I-I FPD compared with the I-T FPDs (ANOVA, Bonferroni, P=0.022). The I-T FPDs experienced statistically significantly more frequent technical failures compared with the other two groups of suprastructures (P=0.003, 0.031). Consequences of complications: The occurrence of loss of retention as a complication increased the odds ratio (OR) to 17.6 (P<0.001) to end up in a technical failure. Similarly, the event of a porcelain fracture increased the OR for the suprastructure to be a failure at 10 years to 11.0 (P< or =0.004). Treatment of periimplantitis increased the OR to 5.44 (P< or =0.011) to result in a biological failure compared with implants in which this type of treatment was not applied. CONCLUSION: The three groups of suprastructures demonstrated marked differences in their patterns of failures and complications. Complications increased the risk for failure. Support by CRF, University of Berne, Switzerland. PMID- 15877754 TI - Economic aspects of single-tooth replacement. AB - The aim of this study was to assess and compare economic parameters of two treatment options in patients requiring single-tooth replacements in private practice. Thirty-seven patients received 41 conventional three-unit fixed partial dentures (FPDs). Fifty-two patients received 59 single crowns on implants (I). Treatment assignment was not random. All except one were metal ceramic reconstructions. All except one were crowns cemented on solid abutments of the ITI((R)) Dental Implant System. Economic parameters were noted for the preparatory phase, the actual reconstruction and for treatment of biological and/or technical complication thereafter (range 1-4 years): number of visits, chair-side time, treatment costs, costs for implant components and laboratory work. Costs were based on the tariffs between the Swiss dentists association and the insurers (Sfr 3.1 per tariff point). Implant treatment required more visits than FPD (8.1+/-2/4.8+/-2.3, chi(2): P=0.02). However, the total treatment time was similar (I: 4.8 h+/-0.9 h/FGM: 5.1 h+/-1.3 h, NS). Laboratory costs were higher for FPD (1527.8+/-209 SFr) vs. 579.6+/-106.9 CHF for I. Costs for treatment of technical and biological complications were similar. Total costs amounted to 3939.4+/-766.4 SFr for FGM vs. 3218+/-512.2 SFr for I (P<0.003, Kolmogorov-Smirnov). Even when considering opportunity costs (50 SFr) for each visit the implant solution was less expensive: 3623.2+/-656.1 SFr vs. 4178.7+/ 822.1 SFr (P<0.04, Kolmogorov-Smirnov). Costs for treatment of complications were similar. In conclusion, over a short observation period, the implant reconstruction demonstrated a more favorable cost/effectiveness ratio. Especially in clinical situations with either non- or minimally restored teeth and sufficient bone, the implant reconstruction is to be recommended from an economical point of view. PMID- 15877755 TI - The sinus lift with phycogenic bone substitute. A histomorphometric study. AB - OBJECTIVES: The aim of this histomorphometric prospective study was to ascertain the efficacy of phycogenic bone substitute in an augmented sinus. The process of graft healing, bone remodeling, and biomaterial replacement was examined. MATERIAL AND METHODS: The phycogenic material (fluorohydroxyapatite) made from calcium-encrusted sea algae was used for the sinus lifts. Twenty-four procedures were carried out (one-stage and two-stage equally) and 45 titanium stepped-screw implants were placed. The patients were followed for 12-23 months. In intervals of 6, 9, 12, or 15 months after the sinus lift, 24 graft specimens were taken with a trephine bur. These specimens were examined histomorphometrically. RESULTS: The grafting material was gradually resorbed and replaced by newly formed bone. Between the sixth and 15th month after the sinus lift, the percentage of newly formed bone grew linearly (from 15.5+/-9.6% to 40.8+/-15.3%) and the percentage of bone substitute decreased linearly (from 34.5+/-8.6% to 13+/-9.6%). After 15 months, the density of trabeculae in grafted bone corresponded to cancellous bone of good quality; however, the bone substitute was not completely resorbed during this period. No significant difference between the quality of the newly formed bone in the cases of the one- and two-stage sinus lifts was found. CONCLUSION: Sinus lift carried out with phycogenic bone substitute was shown to be an effective method with limited invasiveness and a high survival rate of implants (97.8%). PMID- 15877756 TI - Does platelet-rich plasma promote remodeling of autologous bone grafts used for augmentation of the maxillary sinus floor? AB - The aim of this study was to evaluate the effect of platelet-rich plasma (PRP) on remodeling of autologous bone grafts used for augmentation of the floor of the maxillary sinus. In five edentulous patients suffering from insufficient retention of their upper denture related to a severely resorbed maxilla, the floor of both maxillary sinus was augmented with an autologous bone graft from the iliac crest. Randomly, PRP was added to the bone graft used to augment the floor of the left or right sinus (split-mouth design). Three months after the reconstruction, bone biopsies were taken with a trephine from the planned implant sites (N=30). Subsequently, three implants were placed in the left and right posterior maxilla. Microradiograms were made of all biopsies (N=30), whereafter the biopsies were processed for light microscopic examination. In addition, clinical parameters were scored. Wound healing was uneventful, clinically no difference was observed between the side treated with PRP or not. Also microradiographical and histomorphological examination of the biopsies revealed no statistical difference between the PRP- and non-PRP side. One implant placed in the PRP side of the graft was lost during the healing phase. Implant-retained overdentures were fabricated 6 months after implantation. All patients functioned well (follow-up 20.2+/-4.3 months). In this study, no beneficial effect of PRP on wound healing and bone remodeling was observed. It is posed that PRP has no additional value in promoting healing of grafted non-critical size defects. PMID- 15877757 TI - The effect of three different calcium phosphate implant coatings on bone deposition and coating resorption: a long-term histological study in sheep. AB - The present study investigated the hypothesis that hydroxyapatite (HA), tricalcium phosphate (TCP), and a HA-gel coated on endosseous titanium (Ti) implants by spark discharging (SD) and dip coating would achieve predictable osseointegration without evident bioresorption of the coatings on the long term. A costal sheep model was used for the implantation of the HA/SD, HA/TCP/SD, and HA-gel/SD specimens, which were retrieved 6 and 12 months following implantation. HA and Ti coatings on implants obtained by conventional plasma spraying (HA/PS, Ti/PS) were used as controls. Microscopy showed that osseointegration was achieved from all types of implants. No evidence for bioresorption of the HA/SD, HA/TCP/SD, and HA-gel/SD coatings was present but cohesive failure with disruption of the coating/implant interface was seen. A statistical analysis of the histomorphometrical data showed no time-dependent effect, however. HA/PS coatings achieved significantly higher bone-implant contact (BIC) percentages of the total implant surface (toBIC) than the other types of coatings (P=0.01). If the BIC percentages were traced separately for implant portions placed into cortical and cancellous bone (coBIC and caBIC, respectively), detailed analysis showed that the caBIC values of HA-gel/SD and HA/PS coatings were significantly higher than that of the other types of coatings (P=0.01). CaBIC values were highly correlated with toBIC values (P<0.001). The present study showed that the preparation techniques used produced thin, dense, and unresorbable coatings that achieved osseointegration. Compared with the control coatings, however, only HA gel/SD coating can be recommended from the investigated preparation techniques for a future clinical use if a better coating cohesion is achieved. PMID- 15877758 TI - Biodegradation of differently cross-linked collagen membranes: an experimental study in the rat. AB - The aim of the present study was to compare the biodegradation of differently cross-linked collagen membranes in rats. Five commercially available and three experimental membranes (VN) were included: (1) BioGide (BG) (non-cross-linked porcine type I and III collagens), (2) BioMend (BM), (3) BioMendExtend (BME) (glutaraldehyde cross-linked bovine type I collagen), (4) Ossix (OS) (enzymatic cross-linked bovine type I collagen), (5) TutoDent (TD) (non-cross-linked bovine type I collagen, and (6-8) VN(1-3) (chemical cross-linked porcine type I and III collagens). Specimens were randomly allocated in unconnected subcutaneous pouches separated surgically on the back of 40 wistar rats, which were divided into five groups (2, 4, 8, 16, and 24 weeks), including eight animals each. After 2, 4, 8, 16, and 24 weeks of healing, the rats were sacrificed and explanted specimens were prepared for histologic and histometric analysis. The following parameters were evaluated: biodegradation over time, vascularization, tissue integration, and foreign body reaction. Highest vascularization and tissue integration was noted for BG followed by BM, BME, and VN(1); TD, VN(2), and VN(3) showed prolongated, while OS exhibited no vascularization. Subsequently, biodegradation of BG, BM, BME and VN(1) was faster than TD, VN(2), and VN(3). OS showed only a minute amount of superficial biodegradation 24 weeks following implantation. Biodegradation of TD, BM, BME, VN(2), and VN(3) was associated with the presence of inflammatory cells. Within the limits of the present study, it was concluded that cross-linking of bovine and porcine-derived collagen types I and III was associated with (i) prolonged biodegradation, (ii) decreased tissue integration and vascularization, and (iii) in case of TD, BM, BME, VN(2), and VN(3) foreign body reactions. PMID- 15877759 TI - The evaluation of processed cancellous bovine bone as a bone graft substitute. AB - OBJECTIVE: To determine the ability of a novel bovine cancellous bone xenoimplant to act as an osteoconductive graft in an ovine femoral defect model. An autograft harvested from the xenoimplant site was placed in a contralateral limb defect for comparison. MATERIAL AND METHODS: The xenoimplant used had been rendered immunologically inert by a novel defatting and deproteinating process. Following surgical implantation of cores into condylar cancellous bone defects, fluorochrome labels were administered to 12 sheep at 2 1/2, 4 1/2 and 8 weeks. Incorporation of the xenoimplants and autografts into the host bone was compared radiographically and histomorphometrically at 10 weeks. RESULTS: Radiographically, the degree of osteointegration was comparable. Histomorphometric data, consisting of labelled surface (LS) estimates, confirmed osteoconductive properties of both the xenoimplants and autografts. Remodelling activity was greatest in the xenoimplants at 2 1/2, weeks. At 4 1/2 weeks, there was more activity in the autograft, but by 8 weeks they were performing similarly. Xenoimplant-LS estimates were comparable or greater than those of the autograft at all times. CONCLUSIONS: Processed bovine cancellous bone xenoimplants were osteoconductive in this model and show promise for development as a biomaterial in human and veterinary orthopaedic surgery. PMID- 15877762 TI - Annotation: velo-cardio-facial syndrome. AB - BACKGROUND: Velo-cardio-facial syndrome (VCFS), the most frequent known interstitial deletion identified in man, is associated with chromosomal microdeletions in the q11 band of chromosome 22. Individuals with VCFS are reported to have a characteristic behavioural phenotype with high rates of behavioural, psychiatric, neuropsychological and linguistic disorders. METHODS: A selective literature review was undertaken. RESULTS: Children and adults with VCFS have high rates of behavioural, psychiatric and communication disorders. While VCFS children have high rates of ADHD, anxiety and affective disorders, adults have high rates of psychotic disorders, particularly schizophrenia. In addition, the presence of a chromosome 22q11 deletion is associated with specific neuropsychological and neuroanatomical abnormalities. CONCLUSIONS: People with VCFS have a characteristic behavioural phenotype with high rates of behavioural, psychiatric, neuropsychological and communication disorders. Early diagnosis and treatment within a multidisciplinary framework is of paramount importance for VCFS individuals as this will have a major effect in determining the long-term outcome in affected individuals. Longitudinal studies of VCFS children are currently under way to identify precursor symptoms and areas of dysfunction which precede the later development of major psychiatric disorder. Identification of such prodromal features in VCFS may have enormous implications for the clinical management of major psychiatric disorder in VCFS and in the wider population. PMID- 15877763 TI - No effect of MMR withdrawal on the incidence of autism: a total population study. AB - BACKGROUND: A causal relationship between the measles, mumps, and rubella (MMR) vaccine and occurrence of autism spectrum disorders (ASD) has been claimed, based on an increase in ASD in the USA and the UK after introduction of the MMR vaccine. However, the possibility that this increase is coincidental has not been eliminated. The unique circumstances of a Japanese MMR vaccination program provide an opportunity for comparison of ASD incidence before and after termination of the program. METHODS: This study examined cumulative incidence of ASD up to age seven for children born from 1988 to 1996 in Kohoku Ward (population approximately 300,000), Yokohama, Japan. ASD cases included all cases of pervasive developmental disorders according to ICD-10 guidelines. RESULTS: The MMR vaccination rate in the city of Yokohama declined significantly in the birth cohorts of years 1988 through 1992, and not a single vaccination was administered in 1993 or thereafter. In contrast, cumulative incidence of ASD up to age seven increased significantly in the birth cohorts of years 1988 through 1996 and most notably rose dramatically beginning with the birth cohort of 1993. CONCLUSIONS: The significance of this finding is that MMR vaccination is most unlikely to be a main cause of ASD, that it cannot explain the rise over time in the incidence of ASD, and that withdrawal of MMR in countries where it is still being used cannot be expected to lead to a reduction in the incidence of ASD. PMID- 15877764 TI - Genetic and environmental influences on conduct disorder: symptom, domain and full-scale analyses. AB - BACKGROUND: We used variable threshold models which accounted for age and gender differences to investigate the genetic and environmental influences on DSM-IV conduct disorder (CD) at the level of symptoms, aggressive versus non-aggressive domains, and full-scale. METHOD: A community sample of 1100 twin pairs (age 11 18) was interviewed using the Diagnostic Interview Schedule for Children. RESULTS: Behavior genetic model fitting suggested that genetic and environmental influences on individual symptoms varied by symptom. The best-fitting models for aggressive and non-aggressive domains, and full-scale CD included additive genetic effects and unique environmental effects only (AE models). These effects could be constrained across age cohorts and sex. The results suggest that using models that incorporate age- and gender-appropriate thresholds specific to each subject we can account for prevalence differences between cohorts. Heritability estimates were .49, .55 and .53 for the aggressive domain, non-aggressive domain, and full-scales, respectively. These results are in contrast to previous research on antisocial behavior measured with the CBCL reporting higher heritability for aggressive versus non-aggressive domains. CONCLUSIONS: Results suggest that individual symptoms of CD may be differentially heritable. Additionally, CD assessed using DSM-IV criteria may show differing patterns of heritability compared with estimates obtained for other measures of antisocial behavior such as the CBCL. PMID- 15877765 TI - Evidence for substantial genetic risk for psychopathy in 7-year-olds. AB - BACKGROUND: Individuals with early warning signs of life-long psychopathy, callous-unemotional traits (CU) and high levels of antisocial behaviour (AB) can be identified in childhood. We report here the first twin study of high levels of psychopathic tendencies in young children. METHODS: At the end of the first school year, teachers provided ratings of CU and AB for 3687 twin pairs from the Twins Early Development Study (TEDS). For the analyses of extreme CU, we selected same-sex twin pairs where at least one twin scored 1.3 or more standard deviations above the mean on the CU scale (612 probands, 459 twin pairs). For the analysis of extreme AB, we selected same-sex twin pairs where at least one twin scored 1.3 or more standard deviations above the mean on AB scale (444 probands, 364 twin pairs). Furthermore, the extreme AB sample was divided into those who were also extreme on CU (children with psychopathic tendencies; 234 probands, 187 twin pairs) and those who did not score in the extreme for CU (children without psychopathic tendencies; 210 probands, 177 twin pairs). RESULTS: DeFries-Fulker extremes analysis indicated that exhibiting high levels of CU is under strong genetic influence. Furthermore, separating children with AB into those with high and low levels of CU showed striking results: AB in children with high levels of CU is under extremely strong genetic influence and no influence of shared environment, whereas AB in children with low levels of CU shows moderate genetic and shared environmental influence. CONCLUSIONS: The remarkably high heritability for CU, and for AB children with CU, suggests that molecular genetic research on antisocial behaviour should focus on the CU core of psychopathy. Our findings also raise questions for public policy on interventions for antisocial behaviour. PMID- 15877766 TI - Developmental consequences of poor phonological short-term memory function in childhood: a longitudinal study. AB - BACKGROUND: A longitudinal study investigated the cognitive skills and scholastic attainments at 8 years of age of children selected on the basis of poor phonological loop skills at 5 years. METHODS: Children with low and average performance at 5 years were tested three years later on measures of working memory, phonological awareness, vocabulary, language, reading, and number skill. RESULTS: Two subgroups of children with poor early performance on phonological memory tests were identified. In one subgroup, the poor phonological memory skills persisted at 8 years. These children performed at comparable levels to the control group on measures of vocabulary, language and mathematics. They scored more poorly on literacy assessments, but this deficit was associated with group differences in complex memory span and phonological awareness performance. The second subgroup of children performed more highly on phonological memory tests at 8 years, but had enduring deficits in language assessments from 4 to 8 years. CONCLUSIONS: Persistently poor phonological memory skills do not appear to significantly constrain the acquisition of language, mathematics or number skills over the early school years. More general working memory skills do, however, appear to be crucial. PMID- 15877767 TI - The genetic-environmental etiology of parents' perceptions and self-assessed behaviours toward their 5-month-old infants in a large twin and singleton sample. AB - BACKGROUND: Given the importance of parenting for the child's early socio emotional development, parenting perceptions and behaviours, and their correlates, should be assessed as early as possible in the child's life. The goals of the present study were 1) to confirm, in two parallel population-based samples, including a large sample of twins, the factor structure of a new self administered questionnaire assessing both parents' specific parenting perceptions and behaviours toward their 5-month-old infants (i.e., parental self-efficacy, perceived parental impact, parental hostile-reactive behaviours and parental overprotection), 2) to identify the specific risk factors associated with the negative side of these parenting dimensions, 3) to document the genetic environmental etiology of these parenting dimensions through the twin method. METHODS: Parents (2,122 mothers and 1,829 fathers) of 5-month-old infants, and parents of 5-month-old infant twins (510 families) completed the questionnaire (28 items). The data were submitted to a series of confirmatory factor analyses. The contribution to parenting of a variety of risk factors was examined in the two samples using regression analyses. A series of quantitative genetic analyses were performed to quantify the different sources of variation in parenting. RESULTS: A consistent factor structure was found across informants and across samples. There were significant mean differences in parenting between mothers and fathers, as well as between parents of twins and parents of singletons. A differentiated pattern of association with risk factors was found for each dimension of parenting. The twin analyses revealed that shared environment accounted for each parenting dimension. Maternal hostile-reactive behaviours were also moderately related to genetic factors in the child and this association was mainly mediated by the infant difficultness. CONCLUSIONS: The overall pattern of results was consistent with Belsky's (1984) view of parenting as multiply determined. The longitudinal follow-up of these families should provide the means for testing developmental models about the determinants and outcomes of these parenting dimensions. PMID- 15877768 TI - Pre-attack symptomatology and temperament as predictors of children's responses to the September 11 terrorist attacks. AB - BACKGROUND: The aims of this study were to assess the psychological response of children following the September 11, 2001 terrorist attacks in New York and Washington, DC and to examine prospective predictors of children's post-attack responses. METHOD: Children's responses were assessed in a community sample of children in Seattle, Washington, participating in an ongoing study. Symptomatology and temperament assessed prior to the attacks were examined as prospective predictors of post-attack post-traumatic stress (PTS), anxiety, depression and externalizing problems. RESULTS: Children demonstrated PTS symptoms and worries at levels comparable to those in children directly experiencing disasters, with 77% of children reporting being worried, 68% being upset by reminders, and 39% having upsetting thoughts. The most common PTS symptom cluster was re-experiencing, and 8% of children met criteria consistent with PTSD. African-American children reported more avoidant PTS symptoms and being more upset by the attacks than European-American children. Girls reported being more upset than boys. Prior internalizing, externalizing, social competence and self-esteem were related to post-attack PTS; and child inhibitory control, assessed prior to the 9/11 attacks, demonstrated a trend towards an association with post-attack PTS symptoms controlling for prior levels of symptomatology. PTS predicted child-report anxiety and conduct problem symptoms at follow-up, approximately 6 months after 9/11. CONCLUSIONS: Children experiencing a major disaster at a distance or indirectly through media exposure demonstrated worries and PTS symptoms suggesting that communities need to attend to children's mental health needs in response to national or regional disasters. Pre-disaster symptomatology or low self-regulation may render children more vulnerable in response to a disaster, and immediate post-disaster responses predict subsequent symptomatology. These variables might be used in the identification of children in need of intervention. PMID- 15877769 TI - Ethnic group, acculturation, and psychiatric problems in young immigrants. AB - BACKGROUND: The goal of this study was to investigate the effects of ethnic origin and acculturation factors on psychiatric problems among immigrant adolescents. One aim was to examine variations in psychiatric problems according to gender and immigrant generation level. Another aim was to explore ethnic group differences in psychiatric problems and acculturation risk and protective factors. Finally, we examined the potential mediating effect of acculturation in the relationship between ethnic origin and psychiatric symptoms. METHOD: Questionnaire data were collected from 1275 immigrant 10th graders with 11 different ethnic origins. Psychiatric problems were measured by the Strength and Difficulty Questionnaire. Acculturation risk factors involved perceived discrimination and ethnic identity crisis. Protective factors were family values, host and ethnic culture competence. RESULTS: First-generation girls and second generation boys were identified as particularly vulnerable to psychiatric problems. There was significant variation in psychiatric problems and acculturation between ethnic groups. There was substantial ethnic group-level correlation between emotional and conduct problems, and between discrimination and peer problems. Otherwise, a differentiated pattern of high-scoring ethnic groups emerged across the various symptom and acculturation indices. ANOVAs yielded unique effects on each symptom category of both ethnic group and the acculturation risk and protective factors, undermining the notion of a mediating effect of acculturation. CONCLUSIONS: There is a complex pattern of adaptation in cultural context and idiosyncratic relationships between distinct psychiatric symptom groups and socio-cultural factors. Information about the differentiated vulnerability of gender, generation, and ethnic groups to psychiatric morbidity is important to identify groups at special risk, and to produce interventions that are tailored to their needs. Future studies should examine how cultural factors contribute both to resilience and to an increased vulnerability to psychiatric problems. PMID- 15877770 TI - The prototype effect in recognition memory: intact in autism? AB - BACKGROUND: There are two accounts of categorization performance in autism: that there is an impairment in prototype formation (Klinger & Dawson, 2001) and that there is an impairment in processing features held in common between stimuli (Plaisted, O'Riordan, & Baron-Cohen, 1998). These accounts, together with central coherence theory (Frith, 1989; Frith & Happe, 1994), imply a reduced or absent prototype effect in autism. METHOD: Children with autism or Asperger syndrome (n = 15) matched on age, gender, and verbal mental age with typically developing children (n = 15) completed a picture recognition task (Experiment 1). These participants also studied categories of cartoon animals possessing either an average prototype structure (Experiment 2) based on (Younger's 1985) stimuli or a modal structure (Experiment 3) based on (Hayes and Taplin's 1993b) stimuli. Following the study phases, participants completed recognition tests comprising prototypes and other exemplars with varying degrees of similarity to the prototypes. RESULTS: For both participant groups, recognition memory appeared intact (Experiment 1) and a full prototype effect in recognition memory was observed in both Experiment 2 and Experiment 3. CONCLUSIONS: The present studies fail to support predictions of impaired prototype effects in autism. The discussion focuses on key methodological differences between these studies and those that support claims that central coherence, prototype formation, and common feature processing are impaired in autism. PMID- 15877774 TI - Cost of disorders of the brain in Europe. PMID- 15877775 TI - Cost of addiction in Europe. PMID- 15877776 TI - Cost of affective disorders in Europe. PMID- 15877777 TI - Cost of anxiety disorders in Europe. PMID- 15877778 TI - Cost of brain tumour in Europe. PMID- 15877779 TI - Cost of dementia in Europe. PMID- 15877780 TI - Cost of epilepsy in Europe. PMID- 15877781 TI - Cost of migraine and other headaches in Europe. PMID- 15877782 TI - Cost of multiple sclerosis in Europe. PMID- 15877783 TI - Cost of Parkinson's disease in Europe. PMID- 15877784 TI - Cost of psychotic disorders in Europe. PMID- 15877785 TI - Cost of stroke in Europe. PMID- 15877786 TI - Cost of trauma in Europe. PMID- 15877787 TI - Guidelines for the treatment and management of new-onset diabetes after transplantation. AB - Although graft and patient survival after solid organ transplantation have improved markedly in recent years, transplant recipients continue to experience an increased prevalence of cardiovascular disease (CVD) compared with the general population. A number of factors are known to impact on the increased risk of CVD in this population, including hypertension, dyslipidemia and diabetes mellitus. Of these factors, new-onset diabetes after transplantation has been identified as one of the most important, being associated with reduced graft function and patient survival, and increased risk of graft loss. In 2003, International Consensus Guidelines on New-onset Diabetes after Transplantation were published, which aimed to establish a precise definition and diagnosis of the condition and recommend management strategies to reduce its occurrence and impact. These updated 2004 guidelines, developed in consultation with the International Diabetes Federation (IDF), extend the recommendations of the previous guidelines and encompass new-onset diabetes after kidney, liver and heart transplantation. It is hoped that adoption of these management approaches pre- and post-transplant will reduce individuals' risk of developing new-onset diabetes after transplantation as well as ameliorating the long-term impact of this serious complication. PMID- 15877788 TI - The use of low dose octreotide prophylaxis in pancreatic transplants with enteric drainage. Results of a prospective randomized single center trial. AB - AIM: The aim of this study was to investigate the effect of octreotide in the perioperative course of pancreas transplants drained into the bowel in terms of fistula formation, pancreatitis, hemorrhage and thrombosis, and to compare the results to patients not receiving octreotide in a prospectively, randomized single center trial. PATIENTS AND METHODS: Forty pancreas transplant recipients were prospectively randomized to either receive or not to receive octreotide 0.1 mg subcutaneously at the time of operation and 3x/d there after until post operative day 7. The incidence of pancreatic leakage from the anastomosis and the content of peritoneal fluid drainage regarding amylase and lipase concentrations collected by abdominal drains were registered on day 0-10. Both groups were comparable for age, sex, onset of diabetes, surgical procedure and immunosuppressive regimen. RESULTS: There were 35 simultaneous pancreas-kidney transplants and five solitary pancreas transplants, two in the octreotide and three in the control group two pancreas after kidney, one pancreas after liver pancreas, one pancreas after simultaneous pancreas kidney transplantation, one pancreas transplant alone. All had enteric drainage. Twenty patients received octreotide and 20 did not. In one patient, receiving octreotide the pancreas had to be removed for septic complications because of an enteric fistula arising from the anastomosis (1/20 = 5%). The incidence in patients on octreotide vs. non octreotide was 1 vs. 0 for pancreatitis, 2 vs. 3 for hemorrhage, 2 vs. 1 for thrombosis and 2 vs. 0 for pancreatic fistulae resulting in an actual overall 12 months patient survival of 100% in both groups and a pancreas survival of 85% vs. 95%. For primary simulaneous pancreas kidney the pancreas graft survival was 93%. The amylase and lipase concentrations of fluid collections drained into the peritoneum on day 0 to 10 post-operatively indicating pancreatic fistulization was comparable in both groups. CONCLUSION: The use of octreotide following pancreas transplantation did not prevent pancreatic fistula formation from the anastomosis neither from the pancreatic capsule in pancreas transplantation with enteric drainage. Further studies are required to finally evaluate the benefit of this prophylactic treatment. PMID- 15877789 TI - Heart transplant recipient clinical profile improvement following mycophenolate mofetil late incorporation into the treatment schedule. AB - Mycophenolate mofetil (MMF) has a better clinical profile than azathioprine in heart transplantation (HT). Forty-five recipients (aged 53 +/- 9 yr) were retrospectively evaluated (first year of follow-up) post-MMF introduction since its advent in 1997 (mean daily dose: 1.97 +/- 0.2 g). MMF was used (mean post-HT time: 40 +/- 27 months) for: (i) renal insufficiency attenuation (group 1 = 20); (ii) steroid reduction because of osteoporosis (group 2 = 12); (iii) treatment of persistent cellular rejection (group 3 = 7) and vascular graft disease (VGD) (group 4 = 6). Mean changes (groups 1-2) were: creatinine 172 +/- 59, 158 +/- 51, 153 +/- 57 mumol/L (at baseline, 6 and 12 months, respectively; p < 0.001). Cyclosporine daily dose: 219 +/- 37, 166 +/- 46, 176 +/- 98 mg, respectively (p < 0.001). Cyclosporine blood concentration: 151 +/- 40, 103 +/- 41, 83 +/- 34 ng/mL, respectively (p < 0.004). Prednisone daily dose: 8.3 +/- 2, 5.2 +/- 1, 4.1 +/- 1 mg, respectively (p < 0.001). Cellular rejection (group 3) was successfully treated (86%) but the outcome of VGD did not improve after the switch (group 4). Our limited experience (with caution) confirms the reported benefits of MMF particularly attenuating renal insufficiency. PMID- 15877790 TI - A calcineurin antagonist-free induction/maintenance strategy for immunosuppression in elderly recipients of renal allografts from elderly cadaver donors: long-term results from a prospective single centre trial. AB - BACKGROUND: With the aim to improve the inferior outcomes in elderly recipients of kidneys from elderly cadaver donors, we applied and investigated a therapeutic regimen consisting of calcineurin inhibitor (CNI)-free, mycophenolate mofetil (MMF)-based immunosuppressive (i.s.) induction/maintenance protocol. In this article, we report the long-term results of this clinical trial. METHODS: A total of 89 recipients (mean age: 63.8 yr) of kidneys from cadaver donors (mean age: 66.8 yr) were consecutively recruited for this 5-yr, prospective, open, single centre, pilot trial. Induction therapy consisted of MMF and steroids in conjunction with a short course (4-10 d) of rabbit antithymocyte globulin (ATG). Maintenance treatment was performed with MMF/steroids or MMF alone under strict therapeutic drug monitoring by aiming target mycophenolic acid (MPA)-trough levels between 2 and 6 mg/mL. RESULTS: Cumulative 5-year patient and renal allograft survival was 87.69% and 69.81%, respectively. Acute rejection episodes occurred in 23.6% (21 patients). Long-term function of the old renal allografts proved to be satisfactory as reflected by serum creatinine-values of 1.53 mg/dL and urea-values of 57.9 mg/dL at 5 yr. CONCLUSION: Application of a nephrotoxicity- and atherogenicity-free, MMF-based i.s. induction/maintenance protocol in elderly recipient of kidneys from elderly cadaver donors leads to improved long-term outcomes which are comparable with data from young recipients who have received allografts from young cadaver donors. PMID- 15877791 TI - Outcome of renal transplants from pediatric donors <5 yr of age. AB - AIM: Outcomes of single renal transplants from donors <5 yr old have traditionally been inferior to those from older donors. We retrospectively studied our experience with patients who received renal transplants, either individually or en bloc, from young donors (<5 yr of age) to determine the utility of these organs. We also compared the outcomes of these transplant patients maintained on either cyclosporine- (CyA) or tacrolimus-based (TRL) immunosuppression regimens. PATIENTS: Ninety-eight patients received transplants at our center from donors <5 yr of age between August 1993 and August 2003. They were followed-up from 12 months to 11 yr. Patients were divided into four groups based on whether they received single or en bloc transplants, and whether CyA or TRL was the base immunosuppressive agent. Patients in group I (n = 13) received single pediatric kidneys and were treated with CyA regimens; group II patients (n = 26) also received single pediatric kidneys, but were treated with TRL regimens; group III patients (n = 31) were transplanted en bloc and were treated with CyA; and group IV patients (n = 28) received en bloc transplants and were treated with TRL. RESULTS: One-year patient and death-censored graft survival was not significantly different between recipients of en bloc vs. single grafts (i.e. 88 and 85% vs. 90 and 87%, respectively), or between the four treatment groups (group I: 85 and 85%, group II: 92 and 88%, group III: 87 and 84%, and group IV: 89 and 86%, respectively). The overall 1-yr rejection rate was 30% (29 of 98), which was significantly higher in the CyA-treated patients 19 of 44; i.e. 43%, than in TRL-treated patients 10 of 54, i.e. 19%, p = 0.03). In the en bloc recipients, seven grafts (12%) were lost as a result of vascular thrombosis. Notably, none of the single kidneys were lost because of vascular thrombosis. At the end of follow-up the creatinine levels of both groups were comparable. CONCLUSIONS: Pediatric donor kidneys transplanted individually provide for equal patient and graft survival when compared with en bloc transplants. TRL can be used reduce the detrimental effect of acute rejection on graft growth and function when compared with CyA. Single use of such kidneys can safely and efficaciously be transplanted into adult recipients, greatly expanding the donor pool. PMID- 15877792 TI - Effect of lamivudine treatment in patients with decompensated cirrhosis due to anti-HBe positive/HBeAg-negative chronic hepatitis B. AB - Lamivudine has been shown to improve liver function and reduce the need for liver transplantation (LT) in patients with decompensated HBeAg-positive cirrhosis. Nevertheless, there is only limited experience with lamivudine in patients with anti-HBe-positive/HBeAg-negative cirrhosis. The primary aim of this study was to determine whether lamivudine treatment improves liver function and subsequently pre-LT survival or delays or obviates the need for LT in patients with anti-HBe positive/HBeAg-negative cirrhosis. Between July 1998 and June 2003, 20 consecutive patients awaiting LT were enrolled in the study. All patients showed active viral replication and were treated with lamivudine 100 mg daily. Significant clinical improvement, defined as a decrease in the Child-Pugh Turcotte score by >or=2 points, was observed in 11 (55%) patients. The median change in the Child-Pugh-Turcotte score was -2 (range -5 to +2). The median time required to achieve a 2-point or greater reduction in Child-Pugh-Turcotte score was 6 months (range 3-12 months). In nine patients (45%), the Child-Pugh-Turcotte score decreased to or=1 epicardial vessel or any distal vessel attenuation. All patients underwent blood sampling 89 months [67, 119] after HTx for biochemical (glucose, creatinine, total and LDL cholesterol, and cyclosporin levels) and redox evaluation [plasma reduced and total homocysteine, cysteine, cysteinylglycine, glutathione, blood reduced glutathione (GSH(bl)) and vitamin E]. Univariate Odds Ratios (OR) with 95% confidence interval (95% CI, highest vs. lowest quartile) were estimated on the basis of a logistic regression analysis between clinical, conventional biochemical and redox data. Only the significant variables at univariate entered into multivariate analysis. RESULTS: CAV was documented in 15 (27%) patients. Univariate analysis showed that time from HTx to angiography (OR 3.97, 95% CI 1.15-14, p = 0.03) and GSH(bl) (OR 0.31, 95% CI: 0.14-0.70, p = 0.005) were significantly associated with CAV. However, multivariate analysis revealed GSH(bl) as the only independent predictor of CAV (OR 0.31, 95% CI: 0.13-0.74, p = 0.008). CONCLUSIONS: In HTx recipients reduced levels of GSH(bl) are independently associated with CAV. Given its potent intracellular scavenger properties, GSH(bl) may serve as a marker of antioxidant defence consumption, favouring CAV development. PMID- 15877801 TI - Negative impact of 'old-to-old' donations on success of cadaveric renal transplants. AB - The effect of 'old-to-old' cadaveric renal transplants on operative complications and graft survival was assessed in all 325 patients undergoing solitary cadaveric renal transplantations in Israel during a 3-yr period. Preoperative information and hospital course data were abstracted from the charts. Results were analyzed using Kaplan-Meyer survival curves, univariate and multivariate Cox models. Overall, 62 (19.1%) grafts failed within a year. Failure rate was 46.2% for 'old to-old' transplants compared with 15.5% for all other donor/recipient age combinations (p < 0.0001). 'Old-to-old' transplants remained independently associated with graft failure in a multivariate Cox model after controlling the effect of other risk factors. 'Old-to-old' transplants were also associated with increased operative complications relative to other age combinations. The decision to use 'old-to-old' transplants, even when donors are scarce, is problematic and should be reconsidered. PMID- 15877802 TI - Pulmonary infections increase exhaled nitric oxide in lung transplant recipients: a longitudinal study. AB - The aim of this longitudinal study was to test whether pulmonary infections influence fractional exhaled nitric oxide levels (FENO) in otherwise clinically stable lung transplant recipients. Levels of FENO were measured at least on 11 occasions in nine lung transplant recipients who attended for routine or urgent clinical review over 27.0 +/- 3.2 months period. Diagnosis of infection was based on clinical symptoms, functional measurements and radiological findings. Concentrations of FENO were also determined in 12 healthy volunteers. During follow-up, six patients had one, two had three, and one had four episodes of pulmonary infections. Overall, six upper and 10 lower respiratory tract infections were noted. Recipients with active infections developed increased FENO levels as compared with their own baseline levels measured in the clinically well period (10.8 +/- 1.3 vs. 7.6 +/- 1.1 ppb, p < 0.05). After antibiotic treatment, elevated FENO concentrations returned to baseline in association with full clinical recovery. Baseline FENO levels in lung transplant recipients and in healthy volunteers (6.0 +/- 0.5 ppb) were similar. The sensitivity and specificity of FENO measurement in detecting pulmonary infections were 57 and 96%, respectively. Our data suggest that pulmonary infections are associated with increased FENO levels in patients with lung allografts. Nevertheless, the measurement of FENO by itself as a screening tool for infections seems to be limited by its low sensitivity. PMID- 15877803 TI - Expanding the donor pool: use of renal transplants from non-heart-beating donors supported with extracorporeal membrane oxygenation. AB - In response to organ shortage, we used the renal grafts from non-heart-beating donors (NHBDs). Extracorporeal membrane oxygenation (ECMO) was used to maintain NHBDs before organ procurement. We compared the results of renal transplantation from different donors, including heart-beating donors (HBDs), living-related donors (LDs), and NHBDs supported with ECMO. From February 1998 to June 2003, we recruited 219 patients receiving renal transplantation at National Taiwan University Hospital. Among them, 31 received kidneys from NHBDs supported with ECMO, 120 from HBDs, and 68 from LDs. Multiple organ transplant recipients were not included in this study. We compared the graft survival, serum creatinine levels, and estimated glomerular filtration rates of the three groups. The rate of delayed graft function was higher in NHBD recipients (41.9%) than in HBD recipients (27.0%) and LD recipients (10.9%) (p = 0.003). In the NHBD group, the recipients of grafts with delayed function had significantly longer ECMO runs (63.1 +/- 3.0 min) than those without delayed function (53.7 +/- 2.5 min) (p = 0.024). Estimated glomerular filtration rate (p = 0.472) and mean serum creatinine level (p = 0.286) were not significantly different between the three groups using a longitudinal approach. The 5-yr graft survival rates for NHBD (88.4%, 95% CI: 0.680-0.962), HBD (83.2%, 95% CI: 0.728-0.899), and LD transplant recipients (89.3%, 95% CI: 0.619-0.974) were not significantly different (p = 0.239). The 5-yr patient survival rates for NHBD, HBD, and LD transplant recipients were 100, 93.0 (95% CI: 0.859-0.966) and 100% respectively. The long term allograft survival and function of kidneys from NHBDs supported by ECMO, HBD, and LD did not differ significantly. Long ECMO running time tended to delay graft function. PMID- 15877804 TI - Risk factors for delayed kidney function and impact of delayed function on patient and graft survival in adult graft recipients. AB - The influence of delayed kidney graft function on allograft outcome is described controversially in the literature. The aim of the study was to evaluate possible risk factors for delayed graft function (DGF) and investigate the impact of DGF on short- and long-term renal allograft function. Two groups were formed: the first one consisted of patients who gained immediate graft function (IGF) (n = 64) after transplantation and the second group included patients with DGF (n = 31; with at least one dialysis needed in first week after transplantation). The DGF group had a statistically significant longer duration on dialyses prior to transplantation (DGF 54 vs. IGF 33 months; p < 0.05), on average more frequently a re-transplantation (DGF 1.7 vs. IGF 1.3; p < 0.01), a longer re-anastomosis time (DGF 52.9 vs. 44.2 min; p < 0.01), a lower systolic (DGF 136 +/-24 mmHg vs. IGF 158 +/- 25; p < 0.001) and diastolic blood pressure (DGF 78 +/- 14 vs. IGF 89 +/- 16 mmHg; p < 0.01) at admission to the hospital and a higher serum (S) creatinine at discharge (DGF 2.5 +/- 1.6 vs. IGF 1.6 +/- 0.4 mg/dL; p < 0.01). Prior to transplantation the DGF group had more often advanced vascular diseases (DGF 29.0 vs. IGF 12.5%; p < 0.01) and these patients incurred more frequently new ones during the next 3 yr after transplantation (DGF 22.6 vs. IGF 6.3%; p < 0.001). After 3 yr the graft survival tended to be lower in the DGF group (DGF 74.2 vs. IGF 84.4%; NS), but this difference was not statistically significant. PMID- 15877805 TI - Outcome of hepatic artery reconstruction in liver transplantation with an iliac arterial interposition graft. AB - BACKGROUND: In case of anomal hepatic arterial inflow, it can be necessary to perform revascularization of the liver allograft by iliac arterial interposition graft. METHODS: We analyzed retrospectively 613 liver transplants in a 16-yr period. The hepatic artery (HA) graft group (n = 101) consisted of patients with arterial inflow based on recipient infrarenal aorta using donor iliac artery graft tunneled through the transverse mesocolon. The control group (n = 512) consisted of patients who underwent liver transplantation with routine HA reconstruction. RESULTS: Both groups are homogeneous and comparable. In case of retransplantation, arterial conduit with iliac graft was adopted more frequently instead of conventional arterial anastomosis (24.8% vs. 9%, p < 0.0001). The 1-, 3- and 5-yr overall survival was 85.41, 79.42, 76.57% in the control group and 76.21, 73.43, 73.43% in the HA graft group, respectively (p = ns). The 1-, 3- and 5-yr graft survival was better in the control group (81.51, 73.66, 69.22% vs. 71.17, 62.50, 53.42%) (p = 0.01). In case of retransplantation, the 1-, 3- and 5 yr overall (57.81, 53.95, 41.96% vs. 60, 51.95, 49.85%) and graft survival (57.52, 53.68, 41.75% vs. 56, 50.4, 40.3%) was similar in control and HA graft group, respectively (p = ns). Hepatic artery thrombosis (HAT) rate is 21.8% vs. 8.6% (p < 0.0001) in HA graft group and control group, respectively. The only factor independently predictive of early HAT resulted arterial conduit (p = 0.001, OR = 3.13, 95% CI: 1.57-6.21). Retransplant procedure, donor age and arterial iliac conduit were found to be a significant risk factors for late HAT, at univariate analysis. At multivariate analysis, donor age >50 yr old resulted the only factor independently associated with late HAT (p < 0.0001, OR = 1.05, 95% CI: 1.02-1.07). CONCLUSION: Iliac arterial interpositional graft is an alternative solution for arterial revascularization of liver allograft in case of retransplantation when the use of HA is not possible. In case of primary transplantation, is better not to perform arterial conduit if it is possible, for poor graft survival and high incidence of early HAT, especially in case of liver donor aged over 50 yr. PMID- 15877806 TI - United Network for Organ Sharing's expanded criteria donors: is stratification useful? AB - The United Network for Organ Sharing (UNOS) Expanded Criteria Donor (ECD) system utilizes pre-transplant variables to identify deceased donor kidneys with an increased risk of graft loss. The aim of this study was to compare the ECD system with a quantitative approach, the deceased donor score (DDS), in predicting outcome after kidney transplantation. We retrospectively reviewed 49 111 deceased donor renal transplants from the UNOS database between 1984 and 2002. DDS: 0-39 points; >or=20 points defined as marginal. Recipient outcome variables were analyzed by ANOVA or Kaplan-Meier method. There was a 90% agreement between the DDS and ECD systems as predictors of renal function and graft survival. However, DDS identified ECD- kidneys (10.7%) with a significantly poorer outcome than expected (DDS 20-29 points, n = 5,252). Stratification of ECD+ kidneys identified a group with the poorest outcome (DDS >or=30 points). Predictability of early post-transplant events (i.e. need for hemodialysis, decline of serum creatinine and length of hospital stay) was also improved by DDS. DDS predicted outcome of deceased donor renal transplantation better than the ECD system. Knowledge obtained by stratification of deceased donor kidneys can allow for improved utilization of marginal kidneys which is not achieved by the UNOS ECD definition alone. PMID- 15877807 TI - Chronic allograft nephropathy and nephrotic range proteinuria. AB - While the association between post-transplant nephrotic range proteinuria (PTx NP) and chronic allograft nephropathy (CAN) has been described, the factors that determine graft survival in such patients are unclear. We retrospectively identified 30 patients with biopsy-proven CAN who presented with PTX-NP between 1988 and 2002. Patients were stratified into two groups according to PTX-NP onset: <1 yr vs. >1 yr post-transplantation. Both groups were comparable with respect to the degree of renal dysfunction (serum creatinine 4.3 +/- 2.5 mg/dL vs. 3.4 +/- 1.5 mg/dL) and proteinuria (4.7 +/- 1.6 gm/d vs. 5.8 +/- 3 gm/d). After a mean follow-up of 14 months post-biopsy, 87% of patients had lost their grafts in both groups (89% vs. 83%, p = NS). Overall, patients with serum creatinine 2 mg/dL (75% vs. 4%, Fisher Exact Probability p = 0.0038). Using Kaplan-Meier estimate, the 5-yr graft survival rate was 100% for patients with serum creatinine 2 mg/dL (p = 0.06). The magnitude of proteinuria beyond 3 gm/d did not influence graft survival. One-half of the patients (n = 15) received therapy with angiotensin converting enzyme inhibitors (ACEI). Graft survival, however, was not different between the patients who received ACEI compared with the patients who did not receive ACEI (13% vs. 13%). PTx-NP related to CAN was associated with poor allograft survival, irrespective of the time of onset of presentation, especially when renal function was reduced at the time of biopsy. PMID- 15877808 TI - Heparin-induced thrombocytopenia antibody and the pathogenesis of thrombotic microangiopathy after stem cell transplantation. AB - Thrombotic microangiopathy (TMA) that occurs after stem cell transplantation (SCT) is generally regarded as being different from thrombotic thrombocytopenic purpura (TTP), because it is reportedly not associated with deficiency of von Willebrand factor-cleaving protease, whereas this enzyme is deficient in TTP. However, better understanding of the pathogenesis of this condition is still required. Accordingly, we investigated the relationship between TMA occurring after SCT and heparin-induced thrombocytopenia (HIT), a condition related to low dosed heparin therapy that features thrombocytopenia and generalized thrombotic disorders. Thirty-nine consecutive patients who underwent bone marrow transplantation were divided into a TMA group and a non-microangiopathy group (10 and 29 patients, respectively). Before SCT, the serum platelet factor 4 (PF4) levels of the TMA and non-microangiopathy groups were 0.123 +/- 0.023 and 0.132 +/- 0.025, respectively (p = NS). One week after recovery of the white blood cell count following transplantation, the TMA group (0.2902 +/- 0.0678) had a significantly higher PF4 level than the non-microangiopathy group (0.1548 +/- 0.0312) (p < 0.001, t-test). Thus, PF4 increased after engraftment of the transplanted stem cells in the patients who developed TMA. In patients who developed TMA, there was a significant correlation between the PF4 level and the grade of angiopathy according to the Zeigler grading system (p < 0.01 by linear regression analysis). These results suggest that a HIT antibody produced by donor cells may be involved in the development of TMA after SCT. PMID- 15877809 TI - Anti-CD20 monoclonal antibody (Rituximab) for life-threatening hemolytic-uremic syndrome. AB - Rituximab is a chimaeric monoclonal antibody directed against the CD20 antigen. It has been successfully used in B-cell malignancy and its efficacy in the treatment of in autoimmune hemolytic anemia and other autoimmune diseases is being investigated. There are also few case reports of its success in thrombotic thrombocytopenic purpura, but no reports of its use in hemolytic-uremic syndrome (HUS). We report a 36-year-old patient who had lost the function of her native kidneys secondary to HUS. After more than 1 year in clinical remission, she received a living unrelated kidney transplant. This immediately precipitated a severe relapse of HUS. The process was abrogated but not completely inactivated, despite over 40 plasma exchange treatments. Consequently, she was given Rituximab in courses of two to three doses, each dose 375 mg/m(2), at weekly intervals with remarkable stabilization of her disease for approximately 6 months. PMID- 15877810 TI - Is Pneumocystis carinii pneumonia after stem cell transplantations a contagious disease? AB - We report of twins who underwent hematopoietic stem cell transplantation (HSCT) for neonatal acute leukemia. Hospitalized in the same room from the time the first one demonstrated respiratory symptoms, they both developed Pneumocystis jiroveci (formerly carinii) pneumonia (PCP) 2 wk apart. This observation suggests that PCP may be a contagious disease in HSCT recipients. This may be especially true for infants and young children who are at risk of primary P. jiroveci infection, and should be avoided. PMID- 15877811 TI - Phrenic nerve afferents elicited cord dorsum potential in the cat cervical spinal cord. AB - BACKGROUND: The diaphragm has sensory innervation from mechanoreceptors with myelinated axons entering the spinal cord via the phrenic nerve that project to the thalamus and somatosensory cortex. It was hypothesized that phrenic nerve afferent (PnA) projection to the central nervous system is via the spinal dorsal column pathway. RESULTS: A single N1 peak of the CDP was found in the C4 and C7 spinal segments. Three peaks (N1, N2, and N3) were found in the C5 and C6 segments. No CDP was recorded at C8 dorsal spinal cord surface in cats. CONCLUSION: These results demonstrate PnA activation of neurons in the cervical spinal cord. Three populations of myelinated PnA (Group I, Group II, and Group III) enter the cat's cervical spinal segments that supply the phrenic nerve. PMID- 15877812 TI - Adeno-associated virus: from defective virus to effective vector. AB - The initial discovery of adeno-associated virus (AAV) mixed with adenovirus particles was not a fortuitous one but rather an expression of AAV biology. Indeed, as it came to be known, in addition to the unavoidable host cell, AAV typically needs a so-called helper virus such as adenovirus to replicate. Since the AAV life cycle revolves around another unrelated virus it was dubbed a satellite virus. However, the structural simplicity plus the defective and non pathogenic character of this satellite virus caused recombinant forms to acquire centre-stage prominence in the current constellation of vectors for human gene therapy. In the present review, issues related to the development of recombinant AAV (rAAV) vectors, from the general principle to production methods, tropism modifications and other emerging technologies are discussed. In addition, the accumulating knowledge regarding the mechanisms of rAAV genome transduction and persistence is reviewed. The topics on rAAV vectorology are supplemented with information on the parental virus biology with an emphasis on aspects that directly impact on vector design and performance such as genome replication, genetic structure, and host cell entry. PMID- 15877813 TI - Molecular breakpoint cloning and gene expression studies of a novel translocation t(4;15)(q27;q11.2) associated with Prader-Willi syndrome. AB - BACKGROUND: Prader-Willi syndrome (MIM #176270; PWS) is caused by lack of the paternally-derived copies, or their expression, of multiple genes in a 4 Mb region on chromosome 15q11.2. Known mechanisms include large deletions, maternal uniparental disomy or mutations involving the imprinting center. De novo balanced reciprocal translocations in 5 reported individuals had breakpoints clustering in SNRPN intron 2 or exon 20/intron 20. To further dissect the PWS phenotype and define the minimal critical region for PWS features, we have studied a 22 year old male with a milder PWS phenotype and a de novo translocation t(4;15)(q27;q11.2). METHODS: We used metaphase FISH to narrow the breakpoint region and molecular analyses to map the breakpoints on both chromosomes at the nucleotide level. The expression of genes on chromosome 15 on both sides of the breakpoint was determined by RT-PCR analyses. RESULTS: Pertinent clinical features include neonatal hypotonia with feeding difficulties, hypogonadism, short stature, late-onset obesity, learning difficulties, abnormal social behavior and marked tolerance to pain, as well as sticky saliva and narcolepsy. Relative macrocephaly and facial features are not typical for PWS. The translocation breakpoints were identified within SNRPN intron 17 and intron 10 of a spliced non-coding transcript in band 4q27. LINE and SINE sequences at the exchange points may have contributed to the translocation event. By RT-PCR of lymphoblasts and fibroblasts, we find that upstream SNURF/SNRPN exons and snoRNAs HBII-437 and HBII-13 are expressed, but the downstream snoRNAs PWCR1/HBII-85 and HBII-438A/B snoRNAs are not. CONCLUSION: As part of the PWCR1/HBII-85 snoRNA cluster is highly conserved between human and mice, while no copy of HBII-438 has been found in mouse, we conclude that PWCR1/HBII-85 snoRNAs is likely to play a major role in the PWS- phenotype. PMID- 15877814 TI - Characteristics and adaptability of iron- and sulfur-oxidizing microorganisms used for the recovery of metals from minerals and their concentrates. AB - Microorganisms are used in large-scale heap or tank aeration processes for the commercial extraction of a variety of metals from their ores or concentrates. These include copper, cobalt, gold and, in the past, uranium. The metal solubilization processes are considered to be largely chemical with the microorganisms providing the chemicals and the space (exopolysaccharide layer) where the mineral dissolution reactions occur. Temperatures at which these processes are carried out can vary from ambient to 80 degrees C and the types of organisms present depends to a large extent on the process temperature used. Irrespective of the operation temperature, biomining microbes have several characteristics in common. One shared characteristic is their ability to produce the ferric iron and sulfuric acid required to degrade the mineral and facilitate metal recovery. Other characteristics are their ability to grow autotrophically, their acid-tolerance and their inherent metal resistance or ability to acquire metal resistance. Although the microorganisms that drive the process have the above properties in common, biomining microbes usually occur in consortia in which cross-feeding may occur such that a combination of microbes including some with heterotrophic tendencies may contribute to the efficiency of the process. The remarkable adaptability of these organisms is assisted by several of the processes being continuous-flow systems that enable the continual selection of microorganisms that are more efficient at mineral degradation. Adaptability is also assisted by the processes being open and non-sterile thereby permitting new organisms to enter. This openness allows for the possibility of new genes that improve cell fitness to be selected from the horizontal gene pool. Characteristics that biomining microorganisms have in common and examples of their remarkable adaptability are described. PMID- 15877815 TI - Confirmation of human protein interaction data by human expression data. AB - BACKGROUND: With microarray technology the expression of thousands of genes can be measured simultaneously. It is well known that the expression levels of genes of interacting proteins are correlated significantly more strongly in Saccharomyces cerevisiae than those of proteins that are not interacting. The objective of this work is to investigate whether this observation extends to the human genome. RESULTS: We investigated the quantitative relationship between expression levels of genes encoding interacting proteins and genes encoding random protein pairs. Therefore we studied 1369 interacting human protein pairs and human gene expression levels of 155 arrays. We were able to establish a statistically significantly higher correlation between the expression levels of genes whose proteins interact compared to random protein pairs. Additionally we were able to provide evidence that genes encoding proteins belonging to the same GO-class show correlated expression levels. CONCLUSION: This finding is concurrent with the naive hypothesis that the scales of production of interacting proteins are linked because an efficient interaction demands that involved proteins are available to some degree. The goal of further research in this field will be to understand the biological mechanisms behind this observation. PMID- 15877816 TI - The continuing HIV vaccine saga: naked emperors alongside fairy godmothers. AB - The latest developments in the HIV vaccine field were aired at a Keystone Symposium recently. This Commentary summarizes some of the highlights from this meeting, and focuses on some of the developments that appeared particularly promising, as well as those that do not. Unfortunately, the "saga" continues. PMID- 15877817 TI - A combined approach exploring gene function based on worm-human orthology. AB - BACKGROUND: Many aspects of the nematode Caenorhabditis elegans biology are conserved between invertebrates and vertebrates establishing this particular organism as an excellent genetic model. Because of its small size, large populations and self-fertilization of the hermaphrodite, functional predictions carried out by genetic modifications as well as RNAi screens, can be rapidly tested. RESULTS: In order to explore the function of a set of C. elegans genes of unknown function, as well as their potential functional roles in the human genome, we performed a phylogenetic analysis to select the most probable worm orthologs. A total of 13 C. elegans genes were subjected to down-regulation via RNAi and characterization of expression profiles using GFP strains. Previously unknown distinct expression patterns were observed for four of the analyzed genes, as well as four visible RNAi phenotypes. In addition, subcellular protein over-expression profiles of the human orthologs for seven out of the thirteen genes using human cells were also analyzed. CONCLUSION: By combining a whole organism approach using C. elegans with complementary experimental work done on human cell lines, this analysis extends currently available information on the selected set of genes. PMID- 15877818 TI - Vitamin supplementation for prevention of mother-to-child transmission of HIV and pre-term delivery: a systematic review of randomized trial including more than 2800 women. AB - BACKGROUND: Observational studies have suggested that low serum vitamin levels are associated with increased mother-to-child transmission (MTCT) of HIV and increased preterm delivery. We aimed to determine the efficacy of vitamins on the prevention of MTCT and preterm delivery by systematically reviewing the available randomized controlled trials [RCTs]. We conducted systematic searches of 7 electronic databases. We extracted data from the RCTs independently, in duplicate. RESULTS: We included 4 trials in our review. Of the three trials on Vitamin A, two suggested no difference in MTCT, while the third and largest trial (n = 1078) suggested an increased risk of MTCT (Relative Risk 1.35, 95% Confidence Interval [CI], 1.11-1.66, P = 0.009). Two of the vitamin A trials addressed the impact of supplementation on pre-term delivery; one suggested a benefit (RR 0.65, 95% CI, 0.44-0.94) and the other no difference. All three vitamin A trials found no significant effect on infant mortality at 1 year. Of the two trials that looked at multivitamin use, only one addressed the prevention of MTCT, and found a non-significant RR of 1.04 (95% CI, 0.82-1.32). Two of the multivitamin trials found no significant effects on pre-term delivery. The single multivitamin trial examining children's mortality at 1 year yielded a non significant RR of 0.91 (95% CI, 0.17-1.17). CONCLUSION: Randomized trials of vitamins to prevent MTCT have yielded conflicting results without strong evidence of benefit and have failed to exclude the possibility of harm. PMID- 15877819 TI - Retraction: Inactivation of MAP kinase signalling in Myc transformed cells and rescue by LiCl inhibition of GSK3. AB - The corresponding author submitted this article 1 to Molecular Cancer on the assumption that the co-author had agreed to the submission. Since this is not the case, the authors are retracting the article. The corresponding author is deeply sorry for any inconvenience this may have caused to the editorial and publishing staff. An apology is also extended to the readers. PMID- 15877820 TI - Recent developments in the funding and organisation of the New Zealand health system. AB - During the 1990s, the New Zealand health sector went through a decade of turbulence with a series of major structural changes being introduced in a relatively short period of time. The new millennium brought further change, with the establishment of 21 district health boards and the restoration of a less commercially-oriented system. The sector now appears to be more stable. However many incremental changes are in train and there has been considerable turbulence below the surface as key players jostle for position. This paper reports on some of the recent changes that have occurred in the restructuring of the New Zealand health system. Three issues are discussed: the devolution of funds and decision making to district health boards, developments in primary health care, and the position of the private health insurance industry. PMID- 15877821 TI - Tuberculosis masquerading as malignancy: a multimodality approach to the correct diagnosis - a case report. AB - BACKGROUND: Extrapulmonary tuberculosis is one of the great mimickers of medicine, and often masquerades as malignancy. As a result, patients may be referred to oncologists and surgeons for further evaluation and management, delaying the institution of appropriate anti-tuberculous drug therapy. CASE PRESENTATION: We present the case of a 21 year old man with tuberculous osteomyelitis, who was referred to the Bone and Soft Tissue Sarcoma Service at our institution with a provisional diagnosis of malignancy. Further investigation revealed extensive retroperitoneal abdominal and pelvic lymphadenopathy. The recognition of certain patterns on imaging, and finally the isolation of Mycobacterium tuberculosis from tissue samples obtained under image guidance, enabled the correct diagnosis to be made. CONCLUSION: This case highlights the importance of remaining cognisant of the protean manifestations of extrapulmonary tuberculosis, and illustrates the advantage of a clinically directed multi modality imaging approach to diagnosis. PMID- 15877822 TI - Paramagnetic artifact and safety criteria for human brain mapping. AB - Biological effects of magnetic field and their safety criteria, especially effects of gradient magnetic field on the cerebral and pulmonary circulation during functional brain mapping are still unclear. Here we estimated that magnetically induced artifacts for the blood oxygenation level- and flow- based functional magnetic resonance imaging are less than 0.1%, and disturbance in the pulmonary circulation is less than 1.3% even if the field strength of magnetic resonance system is risen up to 10 tesla. These paramagnetic effects are considered to be small and harmless during human brain mapping. PMID- 15877823 TI - An analysis of the use of genomic DNA as a universal reference in two channel DNA microarrays. AB - BACKGROUND: DNA microarray is an invaluable tool for gene expression explorations. In the two-dye microarray, fluorescence intensities of two samples, each labeled with a different dye, are compared after hybridization. To compare a large number of samples, the 'reference design' is widely used, in which all RNA samples are hybridized to a common reference. Genomic DNA is an attractive candidate for use as a universal reference, especially for bacterial systems with a low percentage of non-coding sequences. However, genomic DNA, comprising of both the sense and anti-sense strands, is unlike the single stranded cDNA usually used in microarray hybridizations. The presence of the antisense strand in the 'reference' leads to reactions between complementary labeled strands in solution and may cause the assay result to deviate from true values. RESULTS: We have developed a mathematical model to predict the validity of using genomic DNA as a reference in the microarray assay. The model predicts that the assay can accurately estimate relative concentrations for a wide range of initial cDNA concentrations. Experimental results of DNA microarray assay using genomic DNA as a reference correlated well to those obtained by a direct hybridization between two cDNA samples. The model predicts that the initial concentrations of labeled genomic DNA strands and immobilized strands, and the hybridization time do not significantly affect the assay performance. At low values of the rate constant for hybridization between immobilized and mobile strands, the assay performance varies with the hybridization time and initial cDNA concentrations. For the case where a microarray with immobilized single strands is used, results from hybridizations using genomic DNA as a reference will correspond to true ratios under all conditions. CONCLUSION: Simulation using the mathematical model, and the experimental study presented here show the potential utility of microarray assays using genomic DNA as a reference. We conclude that the use of genomic DNA as reference DNA should greatly facilitate comparative transcriptome analysis. PMID- 15877824 TI - Internal consistency reliability is a poor predictor of responsiveness. AB - BACKGROUND: Whether responsiveness represents a measurement property of health related quality of life (HRQL) instruments that is distinct from reliability and validity is an issue of debate. We addressed the claims of a recent study, which suggested that investigators could rely on internal consistency to reflect instrument responsiveness. METHODS: 516 patients with chronic obstructive pulmonary disease or knee injury participating in four longitudinal studies completed generic and disease-specific HRQL questionnaires before and after an intervention that impacted on HRQL. We used Pearson correlation coefficients and linear regression to assess the relationship between internal consistency reliability (expressed as Cronbach's alpha), instrument type (generic and disease specific) and responsiveness (expressed as the standardised response mean, SRM). RESULTS: Mean Cronbach's alpha was 0.83 (SD 0.08) and mean SRM was 0.59 (SD 0.33). The correlation between Cronbach's alpha and SRMs was 0.10 (95% CI -0.12 to 0.32) across all studies. Cronbach's alpha alone did not explain variability in SRMs (p = 0.59, r2 = 0.01) whereas the type of instrument was a strong predictor of the SRM (p = 0.012, r2 = 0.37). In multivariable models applied to individual studies Cronbach's alpha consistently failed to predict SRMs (regression coefficients between -0.45 and 1.58, p-values between 0.15 and 0.98) whereas the type of instrument did predict SRMs (regression coefficients between 0.25 to -0.59, p-values between <0.01 and 0.05). CONCLUSION: Investigators must look to data other than internal consistency reliability to select a responsive instrument for use as an outcome in clinical trials. PMID- 15877827 TI - Laparoscopic partial nephrectomy for cancer: techniques and outcomes. AB - Open partial nephrectomy is the gold standard nephron-sparing treatment for small renal tumors. Technical aspects of laparoscopic partial nephrectomy have evolved considerably, and the technique is approaching established status at our institution. Over the past 4 years, the senior author has performed more than 400 laparoscopic partial nephrectomies at the Cleveland Clinic. Herein we present our current technique and review contemporary outcome data. PMID- 15877825 TI - BMP-6 inhibits growth of mature human B cells; induction of Smad phosphorylation and upregulation of Id1. AB - BACKGROUND: Bone morphogenetic proteins (BMPs) belong to the TGF-beta superfamily and are secreted proteins with pleiotropic roles in many different cell types. A potential role of BMP-6 in the immune system has been implied by various studies of malignant and rheumatoid diseases. In the present study, we explored the role of BMP-6 in normal human peripheral blood B cells. RESULTS: The B cells were found to express BMP type I and type II receptors and BMP-6 rapidly induced phosphorylation of Smad1/5/8. Furthermore, Smad-phosphorylation was followed by upregulation of Id1 mRNA and Id1 protein, whereas Id2 and Id3 expression was not affected. Furthermore, we found that BMP-6 had an antiproliferative effect both in naive (CD19+CD27-) and memory B cells (CD19+CD27+) stimulated with anti-IgM alone or the combined action of anti-IgM and CD40L. Additionally, BMP-6 induced cell death in activated memory B cells. Importantly, the antiproliferative effect of BMP-6 in B-cells was completely neutralized by the natural antagonist, noggin. Furthermore, B cells were demonstrated to upregulate BMP-6 mRNA upon stimulation with anti-IgM. CONCLUSION: In mature human B cells, BMP-6 inhibited cell growth, and rapidly induced phosphorylation of Smad1/5/8 followed by an upregulation of Id1. PMID- 15877828 TI - Extracorporeal shock wave lithotripsy in the treatment of renal pelvicalyceal stones in morbidly obese patients. AB - INTRODUCTION: Management of urolithiasis in morbidly obese patients is usually associated with higher morbidity and mortality compared to non-obese patients. In morbidly obese patients, since the kidney and stone are at a considerable distance from the skin (compared to non-obese patients) difficulty may be found in positioning the patient so that the stone is situated at the focal point of the lithotripter. OBJECTIVE: To evaluate the outcomes and cost-efficiency of extracorporeal shock wave lithotripsy (ESWL) in the treatment of renal pelvicalyceal stones sized between 6 and 20 mm in morbidly obese patients. MATERIALS AND METHODS: Using various aids, such as mobile overtable module, extended shock pathway and abdominal compression 37 patients with body mass index more than 40 kg/m2 were treated using the Siemens Lithostar-plus third generation lithotripter. The size of renal pelvicalyceal stones was between 6 and 20 mm. Treatment costs for shock wave lithotripsy were calculated. RESULTS: The overall stone free rate at 3 months of 73% was achieved. The mean number of treatments per patient was 2.1. The post-lithotripsy secondary procedures rate was 5.4%. No complications, such as subcapsular haematoma or acute pyelonephritis were recorded. The most effective (87% success rate) and cost-efficient treatment was in the patients with pelvic stones. The treatment of the patients with low caliceal stones was effective in 60% only. The cost of the treatment of the patients with low calyceal stones was in 1.8 times higher than in the patients with pelvic stones. CONCLUSION: We conclude that ESWL with the Siemens Lithostar plus is the most effective and cost-efficient in morbidly obese patients with pelvic stones sized between 6 and 20 mm. 87% success rate was achieved. The increased distance from the skin surface to the stone in those patients does not decrease the success rate provided the stone is positioned in the focal point or within 3 cm of it on the extended shock pathway. ESWL should not be considered as the first line of treatment in the morbidly obese patients with low caliceal stones where the stone was positioned more than 1 cm from the focal point on the extended shock pathway. PMID- 15877829 TI - Retroperitoneoscopy for treatment of renal and ureteral stones. AB - OBJECTIVE: To assess the efficacy of retroperitoneoscopy for treating stones in the renal pelvis and proximal ureter. MATERIALS AND METHODS: In the period from August 2003 to August 2004, 35 retroperitoneoscopies for treatment of urinary stones were performed on 34 patients. Fifteen patients (42%) had stones in the renal pelvis, and in 2 cases, there were associated stones in the upper caliceal group. Twenty patients (58%) had ureteral stones, all of them located above the iliac vessel. Twenty-five patients (71%) had previously undergone at least one session of extracorporeal lithotripsy and 8 patients (26%) also underwent ureteroscopy to attempt to remove the stone. Eight patients underwent retroperitoneoscopy as a primary procedure. Stone size ranged from 0.5 to 6 cm with a mean of 2.1 cm. RESULTS: Retroperitoneoscopy was performed by lumbar approach with initial access conducted by open technique and creation of space by digital dissection. We used a 10-mm Hasson trocar for the optics, and 2 or 3 additional working ports placed under visualization. Following identification, the urinary tract was opened with a laparoscopic scalpel and the stone was removed intact. The urinary tract was closed with absorbable 4-0 suture and a Penrose drain was left in the retroperitoneum. In 17 patients (49%), a double-J stent was maintained postoperatively. Surgical time ranged from 60 to 260 minutes with a mean of 140 minutes. The mean hospital stay was 3 days (1-10 days). The mean length of retroperitoneal urinary drainage was 3 days (1-10 days). There were minor complications in 6 (17.6%) patients and 1 case of conversion due to technical difficulty. Thirty-three patients (94%) became stone free. CONCLUSION: Retroperitoneoscopy is an effective, low-morbidity alternative for treatment of urinary stones. PMID- 15877830 TI - Indications for percutaneous nephrostomy in patients with obstructive uropathy due to malignant urogenital neoplasias. AB - INTRODUCTION: Urogenital neoplasias frequently progress with obstructive uropathy due to local spreading or pelvic metastases. The urinary obstruction must be immediately relieved in order to avoid deterioration in these patients. The percutaneous nephrostomy is a safe and effective method for relief the obstruction; however the indications of such procedures have been questioned in patients with poor prognosis. MATERIALS AND METHODS: A retrospective study was performed with 43 patients (29 female and 14 male) with urogenital neoplasias who were undergoing percutaneous nephrostomy during a 54-month period. The median age was 52 years. The primary tumoral site was the uterine cervix in 53.5% of patients, the bladder in 23.3%, the prostate in 11.6% and other sites in 11.6%. RESULTS: Postoperative complications occurred in 42.3% of the patients. There was no procedure-related mortality. Thirty-nine per cent of the patients died during the hospitalization period due to advanced neoplasia. The mortality rate was higher in patients with prostate cancer (p = 0.006), in patients over 52 years of age (p = 0.03) and in those who required hemodialysis before the procedure (p = 0.02). Thirty-two per cent of the patients survived long enough to undergo some form of treatment focused on the primary tumor. The survival rate was 40% at 6 months and 24.2% at 12 months. The percentage of the lifetime spent in hospitalization was 17.7%. The survival rate was higher in patients with neoplasia of the uterine cervix (p = 0.007) and in patients with 52 years of age or less (p = 0.008). CONCLUSION: Morbidity was high in this patient group; however, the majority of patients could be discharged from hospital and followed at home. Patients under 52 years of age and patients with neoplasia of the uterine cervix benefited most from the percutaneous nephrostomy when compared to patients with hormone therapy-refractory prostate cancer, bladder cancer or over 52 years of age. PMID- 15877831 TI - Surgical complications after renal transplantation in grafts with multiple arteries. AB - INTRODUCTION: Renal transplantation with multiple arteries appears, in literature, associated to a major index of surgical complications. This study compared the surgical complications and short-term outcome renal transplants with multiple arteries and single artery grafts. MATERIALS AND METHODS: The data of 64 renal transplants with multiple arteries performed between January 1995 and December 1999 were compared to the ones of 292 transplants with single renal artery. The aspects analyzed were number of arteries of the graft, donor type, vascular reconstruction technique, the occurrence of surgical complications, the incidence of delayed graft function, graft function 1 month after transplantation, graft loss and the patients' deaths. RESULTS: The incidence of surgical complications in grafts with multiple arteries and single renal artery was respectively: vascular--3.1% and 3.1%; urological--6.3% and 2.7% and other surgical complications--15.6% and 10.6%, respectively. The incidence of lymphoceles was 3.1% in grafts with a single artery and 12.5% in grafts with more than 1 artery (p = 0.0015). The incidence of delayed graft function in grafts with multiple arteries and with a single renal artery was respectively 35.1 and 29.1% (p = 0.295). Mean serum creatinine at the 30th postoperative day was 2.46 and 1.81 in grafts with multiple and with 1 artery, respectively (p = 0.271). CONCLUSIONS: Kidney transplantation using grafts with single and multiple arteries present similar indexes of surgical complications and short-term outcome; lymphoceles were more frequent among grafts with multiple arteries. PMID- 15877832 TI - Repeat prostate biopsies following diagnoses of prostate intraepithelial neoplasia and atypical small gland proliferation. AB - OBJECTIVE: To assess the incidence of diagnosis of high-grade intraepithelial neoplasia or prostate intraepithelial neoplasia (PIN), and atypical small gland proliferation (ASAP) at a uropathology reference center. To assess the indexes and findings on repeat biopsies. MATERIALS AND METHODS: Diagnoses of PIN, ASAP or PIN + ASAP established between January 1, 2001 and December 31, 2003 were searched in our database. We studied repeat biopsies performed up to August 31, 2004. RESULTS: Of 1420 biopsies, ASAP was diagnosed in 26 (1.8%) patients, PIN in 142 (10%) and PIN + ASAP in 40 (2.8%). Repeat biopsies were performed in 98 patients, 16 (61.5%) with ASAP, 53 (37.3%) with PIN and 29 (72.5%) with PIN + ASAP. Carcinoma was diagnosed in 7 cases (43.8%) following a diagnosis of ASAP, 12 (41.4%) of PIN + ASAP and 7 (13.2%) of PIN. The mean interval between repeat biopsies was 299.6 days. There was no difference between groups where cancer was or was not diagnosed on repeat biopsy in relation to age and serum PSA levels. CONCLUSION: Despite explicit recommendations of repeat biopsy on pathology reports and the high incidence of adenocarcinoma on repeat biopsy, re intervention rates following a diagnosis of PIN, ASAP, PIN + ASAP are low in our setting. The diagnosis that most frequently led to repeat biopsy was PIN + ASAP. Adenocarcinoma was most often diagnosed after the initial diagnosis of ASAP. PMID- 15877833 TI - Assessment of serum level of prostate-specific antigen adjusted for the transition zone volume in early detection of prostate cancer. AB - OBJECTIVE: To determine the clinical usefulness of prostate-specific antigen (PSA) density in the transition zone (PSADTZ) for increasing the specificity in early detection of prostate cancer (PCa) and reducing unnecessary biopsies in males with PSA between 4.0 and 10 ng/mL. MATERIALS AND METHODS: This cross sectional study obtained PSADTZ measurements in 68 patients with PSA between 4.0 and 10 ng/mL. All patients underwent transrectal ultrasonography (TRUS) with biopsies. PSADTZ was estimated by dividing the PSA value by the volume of the transition zone (TZ) obtained. We compared performance measurements for these parameters with those from the PSA itself, PSA density (PSAD) and free PSA/total PSA ratio (F/T PSA). The ability of the method in increasing PSA specificity was demonstrated and compared in univariate and multivariate analyses, and by Receiver Operating Characteristic Curves (ROC). RESULTS: Of the 68 patients under study, 17 (25%) were diagnosed with PCa. The TZ volume (p = 0.001) and PSADTZ (p = 0.001) variables presented means that exhibited statistically significant differences. When compared with the area under the curve (AUC), ROC curves obtained by this method revealed that PSADTZ was the strongest predictor for PCa when considering the cut-off point provided by the curve; that is, 0.35 ng/mL/cc. When PSADTZ was employed, the detection failure would be close to 20%, and less than 45% of cases would undergo unnecessary biopsies. On the other hand, when F/T PSA was used, the loss would reach almost 40%; however less than 30% would undergo unnecessary biopsies. Nevertheless, PSADTZ had the only AUC presenting p < 0.05 in significance when compared with 50%, and was consequently discriminative. CONCLUSIONS: PSADTZ increased PSA specificity in early detection of PCa in males with PSA between 4.0 and 10 ng/mL. However, it was shown to have lower predictive value and lower accuracy than the percentage of free PSA since it presents a higher negative predictive value than all other parameters assessed, and it can be considered clinically useful for reducing unnecessary indications for biopsy. PMID- 15877834 TI - Retroperitoneoscopic surgery with extracorporeal uretero-ureteral anastomosis for treating retrocaval ureter. AB - We present a case of retrocaval ureter featuring laparoscopic technique treatment using extraperitoneal access and extracorporeal suture of the ureteral stumps. Surgical time was 130 minutes, and the anastomosis was performed in 40 minutes. There were no intra- or postoperative complications, and the patient was discharged from hospital on the second postoperative day. The medium-term outcome featured similar results to pure laparoscopic technique. We conclude that this technical variation for treatment of retrocaval ureter makes the procedure easier and provides a drastic reduction in surgical time, without compromising the minimally invasive aspect of this kind of approach. PMID- 15877835 TI - Renal cell carcinoma presenting as a cervical mass. AB - The authors report a case of a 60-year-old woman presenting with a renal cell carcinoma in which the first sign leading to its diagnosis was a cervical metastasis, an uncommon site of distant disease in renal neoplasms. The patient had an 18-month history of a progressively enlarging cervical mass at the anterior aspect of the neck. After laboratory and radiological evaluation, the cervical mass was excised, and the microscopic and immunohistochemical patterns suggested the possibility of a metastatic renal cell carcinoma. Computerized tomography of the abdomen showed a solid, 4 cm left renal mass. A radical left nephrectomy was performed, and the histology confirmed the suspected diagnosis. The patient received immunotherapy, and in a follow-up period of 9 months, there was no evidence of recurrent disease. It seems that head and neck metastasis of renal cell carcinoma should preferentially be treated with surgical excision because of the associated morbidity and quality-of-life issues. PMID- 15877836 TI - Von Recklinghausen's disease with urogenital manifestation. AB - Neurofibromatosis or Von Recklinghausen's disease is a rare disease that affects the urogenital system in its visceral form. We report on one case where the patient had a recurring mass located in the vesicouterine space that caused irritative urinary symptoms, which was confirmed as uterine cervix neurofibroma following surgery and immunohistochemical study. We stress the importance of a wide resection of such tumors and the need for long-term follow-up due to the high recurrence index and the risk of malignant transformation. PMID- 15877837 TI - Fibroepithelial polyp of the urethra. AB - The fibroepithelial polyp of the urethra is rare in adults. Hematuria and obstructive urinary symptoms are the most common findings. The treatment of choice is endoscopic resection and the prognosis for these lesions is excellent. There is no previous report on recurrence. We describe 2 new cases, with 1 of them presenting recurrence following surgical resection. PMID- 15877838 TI - Simplified upper pole nephrectomy: initial experience. AB - OBJECTIVE: To assess the results of an upper pole nephrectomy technique on 5 children. MATERIALS AND METHODS: Upper pole nephrectomy was performed on 5 children, including 4 females and 1 male. Age ranged from 3 to 6 years old. The technique was performed without initial dissection of the renal pedicle. The upper pole is incised and removed. Upon its complete dissection, the segment that drains the upper pole is easily identified, clamped and sectioned. RESULTS: Three children with ureterocele and 2 with ectopic ureter underwent this procedure. There was no intra- or postoperative complication with this technique. DSMA scintigraphy showed no decrease in renal function in the remaining kidney following the procedure. CONCLUSION: The polar nephrectomy technique is simple, and has the advantage of not approaching the renal hilum, which makes surgery less laborious and prevents risk of renal damage, hemorrhage and decreased function in the remaining renal portion. PMID- 15877859 TI - Ballooning dispersal using silk: world fauna, phylogenies, genetics and models. AB - Aerial dispersal using silk ('ballooning') has evolved in spiders (Araneae), spider mites (Acari) and in the larvae of moths (Lepidoptera). Since the 17th century, over 500 observations of ballooning behaviours have been published, yet there is an absence of any evolutionary synthesis of these data. In this paper the literature is reviewed, extensively documenting the known world fauna that balloon and the principal behaviours involved. This knowledge is then incorporated into the current evolutionary phylogenies to examine how ballooning might have arisen. Whilst it is possible that ballooning co-evolved with silk and emerged as early as the Devonian (410-355 mya), it is arguably more likely that ballooning evolved in parallel with deciduous trees, herbaceous annuals and grasses in the Cretaceous (135-65 mya). During this period, temporal (e.g. bud burst, chlorophyll thresholds) and spatial (e.g. herbivory, trampling) heterogeneities in habitat structuring predominated and intensified into the Cenozoic (65 mya to the present). It is hypothesized that from the ancestral launch mechanism known as 'suspended ballooning', widely used by individuals in plant canopies, 'tip-toe' and 'rearing' take-off behaviours were strongly selected for as habitats changed. It is contended that ballooning behaviour in all three orders can be described as a mixed Evolutionary Stable Strategy. This comprises individual bet-hedging due to habitat unpredictability, giving an underlying randomness to individual ballooning, with adjustments to the individual ballooning probability being conferred by more predictable habitat changes or colonization strategies. Finally, current methods used to study ballooning, including modelling and genetic research, are illustrated and an indication of future prospects given. PMID- 15877860 TI - Selection of assessment methods for evaluating banana weevil Cosmopolites sordidus (Coleoptera: Curculionidae) damage on highland cooking banana (Musa spp., genome group AAA-EA). AB - Cosmopolites sordidus (Germar) is an important pest on bananas and plantains. Population build-up is slow and damage becomes increasingly important in successive crop cycles (ratoons). Yield loss results from plant loss, mat disappearance and reduced bunch size. Damage assessment requires destructive sampling and is most often done on corms of recently harvested plants. A wide range of damage assessment methods exist and there are no agreed protocols. It is critical to know what types of damage best reflect C. sordidus pest status through their relationships with yield loss. Multiple damage assessment parameters (i.e. for the corm periphery, cortex and central cylinder) were employed in two yield loss trials and a cultivar-screening trial in Uganda. Damage to the central cylinder had a greater effect on plant size and yield loss than damage to the cortex or corm periphery. In some cases, a combined assessment of damage to the central cylinder and cortex showed a better relationship with yield loss than an assessment of the central cylinder alone. Correlation, logistic and linear regression analyses showed weak to modest correlations between damage to the corm periphery and damage to the central cylinder. Thus, damage to the corm periphery is not a strong predictor of the more important damage to the central cylinder. Therefore, C. sordidus damage assessment should target the central cylinder and cortex. PMID- 15877861 TI - The importance of antennae for pea aphid wing induction in the presence of natural enemies. AB - The pea aphid Acyrthosiphon pisum Harris has been shown to produce an increasing proportion of winged morphs among its offspring when exposed to natural enemies, in particular hoverfly larvae, lacewing larvae, adult and larval ladybirds and aphidiid parasitoids. While these results suggest that wing induction in the presence of predators and parasitoids is a general response of the pea aphid, the cues and mechanisms underlying this response are still unclear. Tactile stimuli and the perception of chemical signals as well as visual signals are candidates for suitable cues in the presence of natural enemies. In this paper the hypothesis that the aphids' antennae are crucial for the wing induction in the presence of natural enemies is tested. Antennae of pea aphids were ablated and morph production was scored when aphids were reared either in the presence or the absence of predatory lacewing larvae over a six-day period. Ablation of antennae resulted in a drastic drop in the proportion of winged morphs among the offspring, both in the presence and the absence of a predator whereas predator presence increased wing induction in aphids with intact antennae, as reported in previous experiments. The results show that antennae are necessary for wing induction in the presence of natural enemies. Critical re-examination of early work on the importance of aphid antennae and tactile stimuli for wing induction suggests that a combination of tactile and chemical cues is likely to be involved not only in predator-induced wing formation but also for wing induction in response to factors such as crowding in the aphid colony. PMID- 15877862 TI - Interactions between the solitary endoparasitoid, Meteorus gyrator (Hymenoptera: Braconidae) and its host, Lacanobia oleracea (Lepidoptera: Noctuidae), infected with the entomopathogenic microsporidium, Vairimorpha necatrix (Microspora: Microsporidia). AB - Infection of Lacanobia oleracea (Linnaeus) larvae with the microsporidium Vairimorpha necatrix (Kramer) resulted in significant effects on the survival and development of the braconid parasitoid, Meteorus gyrator (Thunberg). Female M. gyrator did not show any avoidance of V. necatrix-infected hosts when they were selecting hosts for oviposition. When parasitism occurred at the same time as infection by the pathogen, or up to four days later, no significant detrimental effects on the parasitoid were observed. However, when parasitism occurred six to eight days after infection, a greater proportion (12.5-14%) of hosts died before parasitoid larvae egressed. Successful eclosion of adult wasps was also reduced. When parasitism and infection were concurrent, parasitoid larval development was significantly faster in infected hosts, and cocoons were significantly heavier. However, as the time interval between infection and parasitism increased, parasitoid larval development was significantly extended by up to two days, and the cocoons formed were significantly (c. 20%) smaller. Vairimorpha necatrix spores were ingested by the developing parasitoid larvae, accumulated in the occluded midgut, and were excreted in the meconium upon pupation. PMID- 15877863 TI - How host larval age, and nutrition and density of the parasitoid Dinarmus basalis (Hymenoptera: Pteromalidae) influence control of Acanthoscelides obtectus (Coleoptera: Bruchidae). AB - Choice of the targeted host developmental stage, regulation of parasitoid numbers released and introduction of food supplements are operational factors with a potential to influence the level of biological control. In a closed laboratory storage system maintained over two generations of the host, the impact of these three parameters on the control potential of the parasitoid Dinarmus basalis Rondani was investigated for high populations of larvae of Acanthoscelides obtectus (Say) feeding inside dry common bean seeds Phaseolus vulgaris. The beans were already infested with immature bruchids at the beginning of the storage period to simulate harvest conditions, characterized in a previous study. Treatments resulted in a reduction of 48-75% of the bruchid population within 16 weeks of storage. The best timing of parasitoid release was at the simulated harvest, as later releases reduced the bruchid population only by about half this percentage. Host feeding is postulated to be the key factor involved in the observed difference. The effect of increasing the number of parasitoids strongly depended on host age and food supplement. Addition of vials with honey had no direct effect on the bruchid population or on the parasitoid progeny. The ecological significance of these findings and implications for biological control are discussed. PMID- 15877864 TI - The effect of environmentally induced changes in the bark of young conifers on feeding behaviour and reproductive development of adult Hylobius abietis (Coleoptera: Curculionidae). AB - Young plants of Sitka spruce, Scots and Corsican pine were subject to high and low light, and high and low nitrogen treatments in a polyhouse experiment. The effect of treatments on resin duct size and nitrogen concentration in stem bark was determined together with feeding by Hylobius abietis Linnaeus on the stems of 'intact' plants and on 'detached' stems cut from the plant. Resin duct size was largest on Corsican pine and smallest on Sitka spruce and inherent variation in duct size between the three conifer species appears to determine the pattern of weevil feeding between species. Resin ducts and the flow of resin from them protect the stems of young conifers from weevil feeding not by affecting the total amount of bark eaten but by limiting the depth of feeding and so protecting the inner phloem and cambium. Shallow feeding may increase the likelihood of effective wound repair. Duct size was positively related to plant growth and in particular increased with bark thickness. Overall, ducts were largest in the high light treatment although species differed in their response to the treatment. It is suggested that the effects of plant size, growing conditions and transplantation on susceptibility to attack by H. abietis, reported in various studies, may be due to underlying variation in resin duct size or flow rate. The effect on weevils of superficial feeding on stems is to increase the time for reproductive maturation by reducing consumption of the inner bark which has a higher nitrogen content. PMID- 15877865 TI - Monoclonal antibodies reveal the potential of the tetragnathid spider Pachygnatha degeeri (Araneae: Tetragnathidae) as an aphid predator. AB - The drive towards a more sustainable and integrated approach to pest management has engendered a renewed interest in conservation biological control, the role of natural enemy communities and their interactions with prey. Monoclonal antibodies have provided significant advances in enhancing our knowledge of trophic interactions and can be employed to help quantify predation on target species. The tetragnathid spider Pachygnatha degeeri Sundevall was collected from fields of winter wheat in the UK and assayed by ELISA for aphid proteins. It was demonstrated that this spider did not simply consume greater quantities of aphids because it was bigger. In addition, P. degeeri contained significantly greater concentrations of aphid in their guts than other spiders, showing that aphids comprised a greater proportion of their diet. Although P. degeeri constituted only 6% of the spider population numerically, females and males respectively contained 16% and 37% of total aphid proteins within all spiders screened, significantly more than their density would predict. These spiders also preyed upon aphids at a disproportionately high rate in June, during the aphid establishment phase, theoretically the best time for limiting growth in the aphid population. Although less abundant than other generalist predators, the capability of these hunting spiders to consume large numbers of aphids highlights them as a more significant component of the predator complex than had previously been realized. Limitation of aphid numbers early in the year by generalist predators provides more time for the specialist aphid predators and parasitoids to move in. PMID- 15877866 TI - Abundance, dispersion and parasitism of the stem borer Busseola fusca (Lepidoptera: Noctuidae) in maize in the humid forest zone of southern Cameroon. AB - This study was conducted in the humid forest zone of Cameroon, in 2002 and 2003. The main objective was to investigate the effects of intercropping on infestation levels and parasitism of the noctuid maize stem borer Busseola fusca Fuller. Two trials were planted per year, one during the long and one during the short rainy season. Maize monocrops were compared with maize/legume or maize/cassava intercrops in two spatial arrangements: maize on alternate hills or in alternate rows. Spatial analyses showed that the stemborer egg batches were regularly dispersed in the maize monocrop and aggregated in the intercrops, as indicated by b, the index of dispersion of Taylor's power law. Depending on the crop association and planting pattern, intercrops reduced the percentage of plants with stem borer eggs by 47.4-58.4% and egg densities by 41.2-54.5% compared to monocropped maize. Consequently, larval densities were 44.4-61.5% lower in intercrops compared to monocrops. Intercropping maize with non-host plants did not affect larval parasitism. Up to two-fold higher levels of egg parasitism by scelionid Telenomus spp. were recorded in inter- compared to monocrops during the short rainy seasons of 2002 and 2003. No differences were found among the mixed cropping treatments and parasitism was lower during the long compared to the short rainy seasons. It was proposed that differences in levels of parasitism were due to density dependence effects rather than the effect of the presence of non-host plants in the system. PMID- 15877867 TI - Paraoxonase 1 Q192R (PON1-192) polymorphism is associated with reduced lipid peroxidation in healthy young men on a low-carotenoid diet supplemented with tomato juice. AB - The HDL-bound enzyme paraoxonase (PON) protects LDL from oxidation and may therefore attenuate the development of atherosclerosis. We examined the effect of tomato and carrot juice consumption on PON1 activity and lipid peroxidation in healthy young volunteers with different PON1-192 genotypes (Q/R substitution at position 192). In this randomized cross-over study twenty-two healthy, non smoking men on a low-carotenoid diet received 330 ml/d tomato juice (37.0 mg lycopene, 1.6 mg beta-carotene) or carrot juice (27.1 mg beta-carotene, 13.1 mg alpha-carotene) for 2 weeks. Intervention periods were preceded by 2-week low carotenoid intake. We determined the PON1-192 genotype by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) and measured ex vivo LDL oxidation (lag time), plasma malondialdehyde and PON1 activity at the beginning and end of each intervention period. At baseline, lag time was higher (P<0.05) in QQ (111 (sd 9) min) than in QR/RR subjects (101 (sd 8) min). Neither tomato nor carrot juice consumption had significant effects on PON1 activity. However, tomato juice consumption reduced (P<0.05) plasma malondialdehyde in QR/RR (Delta: -0.073 (sd 0.11) micromol/l) as compared to QQ subjects (Delta:+0.047 (sd 0.13) micromol/l). Carrot juice had no significant effect on malondialdehyde irrespective of the PON1-192 genotype. Male volunteers with the QR/RR genotype showed an increased lipid peroxidation at baseline. Although tomato and carrot juice fail to affect PON1 activity, tomato juice intake reduced lipid peroxidation in healthy volunteers carrying the R-allele of the PON1-192 genotype and could thus contribute to CVD risk reduction in these individuals. PMID- 15877868 TI - Age-related alteration of vitamin D metabolism in response to low-phosphate diet in rats. AB - The responses of renal vitamin D metabolism to its major stimuli alter with age. Previous studies showed that the increase in circulating 1,25-dihydroxyvitamin D (1,25(OH)2D3) as well as renal 25-hydroxyvitamin D3 1-alpha hydroxylase (1-OHase) activity in response to dietary Ca or P restriction reduced with age in rats. We hypothesized that the mechanism involved in increasing circulating 1,25(OH)2D3 in response to mineral deficiency alters with age. In the present study, we tested the hypothesis by studying the expression of genes involved in renal vitamin D metabolism (renal 1-OHase, 25-hydroxyvitamin D 24-hydroxylase (24-OHase) and vitamin D receptor (VDR)) in young (1-month-old) and adult (6-month-old) rats in response to low-phosphate diet (LPD). As expected, serum 1,25(OH)2D3 increased in both young and adult rats upon LPD treatment and the increase was much higher in younger rats. In young rats, LPD treatment decreased renal 24-OHase (days 1-7, P<0.01) and increased renal 1-OHase mRNA expression (days 1-5, P<0.01). LPD treatment failed to increase renal 1-OHase but did suppress 24-OHase mRNA expression (P<0.01) within 7 d of LPD treatment in adult rats. Renal expression of VDR mRNA decreased with age (P<0.001) and was suppressed by LPD treatment in both age groups (P<0.05). Feeding of adult rats with 10 d of LPD increased 1 OHase (P<0.05) and suppressed 24-OHase (P<0.001) as well as VDR (P<0.05) mRNA expression. These results indicate that the increase in serum 1,25(OH)2D3 level in adult rats during short-term LPD treatment is likely to be mediated by a decrease in metabolic clearance via the down-regulation of both renal 24-OHase and VDR expression. The induction of renal 1-OHase mRNA expression in adult rats requires longer duration of LPD treatment than in younger rats. PMID- 15877869 TI - Nutritional regulation of proteases involved in fetal rat insulin secretion and islet cell proliferation. AB - Epidemiological studies have indicated that malnutrition during early life may programme chronic degenerative disease in adulthood. In an animal model of fetal malnutrition, rats received an isoenergetic, low-protein (LP) diet during gestation. This reduced fetal beta-cell proliferation and insulin secretion. Supplementation during gestation with taurine prevented these alterations. Since proteases are involved in secretion and proliferation, we investigated which proteases were associated with these alterations and their restoration in fetal LP islets. Insulin secretion and proliferation of fetal control and LP islets exposed to different protease modulators were measured. Lactacystin and calpain inhibitor I, but not isovaleryl-L-carnitine, raised insulin secretion in control islets, indicating that proteasome and cysteinyl cathepsin(s), but not mu calpain, are involved in fetal insulin secretion. Insulin secretion from LP islets responded normally to lactacystin but was insensitive to calpain inhibitor I, indicating a loss of cysteinyl cathepsin activity. Taurine supplementation prevented this by restoring the response to calpain inhibitor I. Control islet cell proliferation was reduced by calpain inhibitor I and raised by isovaleryl-L carnitine, indicating an involvement of calpain. Calpain activity appeared to be lost in LP islets and not restored by taurine. Most modifications in the mRNA expression of cysteinyl cathepsins, calpains and calpastatin due to maternal protein restriction were consistent with reduced protease activity and were restored by taurine. Thus, maternal protein restriction affected cysteinyl cathepsins and the calpain-calpastatin system. Taurine normalised fetal LP insulin secretion by protecting cysteinyl cathepsin(s), but the restoration of LP islet cell proliferation by taurine did not implicate calpains. PMID- 15877870 TI - Novel tempeh (fermented soyabean) isoflavones inhibit in vivo angiogenesis in the chicken chorioallantoic membrane assay. AB - Anti-angiogenic strategies are emerging as an important tool for the treatment of cancer and inflammatory diseases. In the present investigation we isolated several isoflavones from a tempeh (fermented soyabean) extract. The isolated isoflavones were identified as 5,7,4'-trihydroxyisoflavone (genistein), 7,4' dihydroxyisoflavone (daidzein), 6,7,4'-trihydroxyisoflavone (factor 2), 7,8,4' trihydroxyisoflavone (7,8,4'-TriOH) and 5,7,3',4'-tetrahydroxyisoflavone (orobol). The effects on angiogenesis of these isoflavones were evaluated in the chicken chorioallantoic membrane assay; their capacity to inhibit vascular endothelial growth factor-induced endothelial cell proliferation and expression of the Ets 1 transcription factor, known to be implicated in the regulation of new blood vessel formation, were also investigated. We found that all isoflavones inhibited angiogenesis, albeit with different potencies. Compared with negative controls, which slightly inhibited in vivo angiogenesis by 6.30 %, genistein reduced angiogenesis by 75.09 %, followed by orobol (67.96 %), factor 2 (56.77 %), daidzein (48.98 %) and 7,8,4'-TriOH (24.42 %). These compounds also inhibited endothelial cell proliferation, with orobol causing the greatest inhibition at lower concentrations. The isoflavones also inhibited Ets 1 expression, providing some insight into the molecular mechanisms of their action. Furthermore, the chemical structure of the different isoflavones suggests a structure-activity relationship. Our present findings suggest that the new isoflavones might be added to the list of low molecular mass therapeutic agents for the inhibition of angiogenesis. PMID- 15877871 TI - Hyperinsulinaemia, supplemental protein and branched-chain amino acids when combined can increase milk protein yield in lactating sows. AB - The aim of this study was to determine whether dietary supplementation with branched-chain amino acids, and the infusion of insulin and dextrose, would increase milk protein secretion in the sow. The experiment involved sixteen lactating sows fed either a normal lactation diet (162 g/kg crude protein, n 8) or a high-protein diet (230 g/kg crude protein, n 8) supplemented with branched chain amino acids (valine, isoleucine and leucine). Sows were either infused with insulin and dextrose or not infused at all during mid (day 5-10) and late (day 17 22) lactation in a single reversal design. Blood samples were analysed for glucose, and the dextrose infusion rate was adjusted to maintain the blood glucose level within 15 % of pre-infusion levels. Milk (10.1 v. 11.1 kg/d; P=0.014) and lactose (628 v. 727 g/d; P=0.002) yield increased with insulin infusion, whereas milk protein content (5.0 % v. 5.5 %; P=0.007) was increased in diets supplemented with protein and branched-chain amino acids. Piglet growth was increased by feeding the higher-protein diet (237 v. 273 g/d; P=0.05) but not significantly increased by insulin infusion (245 v. 265 g/d; P=0.11). These effects were additive such that the combined treatment resulted in a 24 % (56 g/d; P<0.05) increase in piglet growth rate. These data demonstrate that increasing the dietary protein/branched-chain amino acid content can increase milk protein secretion but not milk yield. The infusion of insulin and dextrose increased milk and milk lactose yields, and tended to increase milk protein yield but not milk protein content. These effects are additive and translate to increased protein yield and piglet growth. PMID- 15877872 TI - Vitamin E supplementation, cereal feed type and consumer sensory perceptions of poultry meat quality. AB - Lipid oxidation leads to meat spoilage and has been reported to cause adverse changes in the flavour and texture of poultry meat. Vitamin E has been found to be effective in delaying lipid oxidation. The aim of this study was to determine whether the vitamin E supplementation of chicken feed influences the consumers' perception of the quality of chicken meat under normal display and storage conditions. Untrained consumers (n 32) evaluated cooked breast meat from chickens (both corn fed and wheat fed) supplemented with 75,250 or 500 mg/kg vitamin E and after storage at 4 degrees C for 4 and 7 d. Factorial analysis found an interaction between vitamin E treatment and storage day upon the perceived juiciness (P=0.023) and tenderness (P=0.041) of the chicken meat. Perceptions of quality relative to vitamin E level were more evident on day 4 than day 7. When the two cereal types were compared, the time-related sub-group effects were observed only in meat from corn-fed chickens supplemented with either 75 or 250 mg/kg, which was perceived to be juicier (P=0.018) and more tender (P=0.020) than that supplemented at the 500 mg/kg level. These results imply that the two lower concentrations of vitamin E have some advantages over 500 mg/kg, but for optimal consumer acceptance of corn-fed chicken meat, we suggest that 250 mg/kg vitamin E should be added to corn-fed poultry feed. There was no evidence to suggest any advantages in changing the current amount of vitamin E (75 mg/kg) used to rear wheat-fed birds. PMID- 15877873 TI - Bone remodelling is not affected by consumption of a sodium-rich carbonated mineral water in healthy postmenopausal women. AB - This study was designed to investigate the possible effects of consuming Na-rich carbonated mineral water on bone remodelling and urinary mineral excretion in postmenopausal women. Women (n 18) included were amenorrhoeic (>1 year), healthy and not obese (BMI <30 kg/m2). No woman was taking oestrogen replacement therapy, mineral and vitamin supplements, phyto-oestrogens or medications known to affect bone and lipid metabolism. In two consecutive interventions that lasted 8 weeks each, women drank 1 litre of control mineral water daily and 1 litre of carbonated mineral water, rich in Na, HCO3- and Cl-, daily. Body weight and height were measured, BMI was calculated and blood pressure was measured. Blood samples were taken from fasting subjects and serum obtained to analyse the biochemical bone markers, procollagen I amino-terminal propeptide (PINP) and beta carboxy-terminal telopeptide of collagen (beta-CTX). At the end of each period, 24 h urine samples were collected to determine Ca, Mg, P, Na+, K+, Cl-, urine excretion and urinary pH. No changes in body weight, BMI or blood pressure were observed during the experimental period. Ca excretion was lower after the intake of carbonated water than after intake of the control water (P=0.037) while P excretion was higher (P=0.015). Total urine, Na and Cl- excretion did not differ between the two periods but urinary pH was increased after the intake of carbonated mineral water. PINP and beta-CTX did not differ between the two periods. Daily consumption of 1 litre of Na-rich carbonated mineral water for 8 weeks does not affect bone remodelling in healthy postmenopausal women. PMID- 15877874 TI - Elements of Mediterranean diet improve oxidative status in blood of kidney graft recipients. AB - Patients were fully informed as to the study objectives and benefits, and provided written consent prior to enrolment. The study protocol was approved by the Committee on Human Research at the Pomeranian Medical University, Szczecin, Poland. An intensification of free-radical reactions may contribute to accelerated atherosclerosis in kidney graft recipients. We examined the effect of a Mediterranean-type diet (MD) on the oxidative status of the plasma and erythrocytes of kidney graft recipients. Two patient groups were formed: a study group consuming the MD diet and a control group with a low-fat diet. C-reactive protein levels in plasma, oleic acid C18 : 1n-9 and linoleic acid C18 : 2n-6 concentrations in triacylglycerols were determined. To determine the oxidative status, we measured the concentrations of alpha-tocopherol in plasma, the content of thiobarbituric acid-reactive species (TBARS) in plasma and erythrocytes, and the activities of superoxide dismutase, catalase and glutathione peroxidase in erythrocytes. In the MD group, the activities of erythrocyte enzymes changed significantly: those of superoxide dismutase increased (P<0.001 after 6 months), catalase decreased (P<0.001 after 6 months) and glutathione peroxidase decreased (P<0.05 after 2 months). The oleic acid content of triacylglycerols was increased (P<0.006) whereas that of linoleic acid was decreased (P<0.00005), alpha tocopherol levels remaining unchanged. TBARS in plasma were decreased after 6 months of MD (P<0.05). No significant correlations were observed between TBARS, oleic acid, linoleic acid and alpha-tocopherol levels in plasma. MD appears to protect the erythrocytes against the action of free radicals, as reflected in the modified activities of some enzymes regulating the oxidative status of these blood cells. PMID- 15877875 TI - Aleurone flour increases red-cell folate and lowers plasma homocyst(e)ine substantially in man. AB - Aleurone flour (ALF) is a rich source of natural folate (>500 microg/100 g wet weight). Our objective was to establish whether intake of ALF in man can significantly improve folate status and reduce plasma homocyst(e)ine. We performed a randomised, controlled intervention, of 16 weeks duration, in free living healthy individuals (mean age 46-52 years). Participants were assigned to one of three groups: ALF, 175 g bread made with ALF and placebo tablet each day; PCS, 175 g bread made with pericarp seed coat (PCS) flour and placebo tablet each day (low-folate control); or FA, 175 g bread made with PCS flour and tablet containing 640 microg folic acid each day (high-folate control). The daily folate intake contributed by the bread and tablet was 233 microg in the PCS group, 615 microg in the ALF group and 819 microg in the FA group. The number of participants completing all phases of the PCS, ALF and FA interventions was twenty-five, twenty-five and eighteen, respectively. Plasma and red-cell folate increased significantly (P<0.0001) and plasma homocyst(e)ine decreased significantly (P<0.0001) in the ALF and FA groups only. Plasma folate and red cell folate in the ALF group (mean, 95 % CI) increased from baseline values of 12.9 (9.9, 15.7) nmol/l and 509 (434, 584) nmol/l to 27.1 (22.5, 31.7) nmol/l and 768 (676, 860) nmol/l, respectively. Plasma homocyst(e)ine in the ALF group decreased from 9.1 (8.2, 10.0) micromol/l at baseline to 6.8 (6.2, 7.5) micromol/l after 16 weeks. In conclusion, moderate dietary intake of ALF can increase red-cell folate and decrease plasma homocyst(e)ine substantially. PMID- 15877876 TI - Eight-year trends in food, energy and macronutrient intake in a sample of elderly German subjects. AB - Time trends in the food and the corresponding energy and macronutrient intake between 1994 and 2002 are investigated as part of the longitudinal study on nutrition and health status in an ageing population in Giessen, Germany (GISELA study). The dietary intake is assessed via a 3 d estimated dietary record especially developed and validated for this study. A total of 2135 records from 532 subjects, aged 60 years and over, are analysed separately according to sex and age groups (born 1904-1928 and born 1929-1942). Results show that the food, energy and macronutrient intake of the GISELA subjects stayed fairly stable over the study period. However, some significant changes in the consumption pattern are observed, predominantly in the women and the younger age groups, which correspond to the national trends in Germany. Most obvious changes observed among the GISELA subjects are the increased intake of water (between +21.05 and +41.61 ml/d per year) and vegetables (between +2.94 and +3.38 g/d per year) and a reduced consumption of coffee and tea (between -10.65 and -15.99 ml/d per year) as well as of meat (between -1.79 and -2.56 g/d per year). The healthier food choices and the ability to change food habits could be related to the relatively high health-awareness of the GISELA subjects compared to the federal average of equivalent age groups. PMID- 15877877 TI - The Diet Quality Index-International (DQI-I): is it a useful tool to evaluate the quality of the Mediterranean diet? AB - The aim of this study was to assess whether the recently developed Diet Quality Index-International (DQI-I) could be used to evaluate diet quality of a Mediterranean population. A cross-sectional nutritional survey was carried out in the Balearic Islands (Spain) between 1999 and 2000. Dietary information (replicated 24 h recall and a food frequency questionnaire), and socio demographic and lifestyle data were collected from a representative sample of the population (n 1200: 498 males and 702 females) aged 16-65 years (response rate 77.22 %). The DQI-I was developed according to the method defined by Kim et al. (2003), and focused on four major aspects of a high-quality diet (variety, adequacy, moderation and overall balance). The percentage of adherence to the Mediterranean dietary pattern (MDP) was also calculated and correlation analysis was carried out between the DQI-I score and the percentage of adherence to the MDP. The total score of the DQI-I reached 43 % of the possible score, indicating that the Balearic diet was a poor-quality diet. Correlation analysis between the DQI-I scores and adherence to the MDP showed that the DQI-I subcategories protein, iron and calcium adequacy were negatively correlated with the MDP. Furthermore, moderation in empty calorie food consumption and overall balance subcategories were not significantly correlated with the MDP. Due to some methodological factors and cultural biases, the proposed DQI-I scoring system is not useful to evaluate the quality of this Mediterranean-type diet. Further research is needed to develop a new diet quality index adapted to the MDP. PMID- 15877878 TI - Simulation of prospective phytosterol intake in Germany by novel functional foods. AB - A blood cholesterol-lowering margarine containing plant sterolesters was the first functional food placed on the European food market pursuant to the regulation (EC) 258/97. In the following years nine further applicants submitted the request to add plant sterol compounds to dairy products, cheeses, bakery products, sausages, plant oils and other products. The European Scientific Committee on Food (SCF) declared a precautionary intake limit of 3 g plant sterols per d by multiple dietary sources. Using the consumption data of the German National Food Consumption Study, carried out from 1985 to 1988 with 23 209 participants, we hypothetically added 0.3-2 g plant sterols to usual daily servings of ten different food products, selected from the novel food applications. We calculated the prospective plant sterol intake regarding each kind of enriched food and by stepwise accumulation of different functional foods in three enrichment scenarios. Within our enrichment context we find a phytosterol intake satiation, if multiple plant sterol-enriched foods are eaten. An enrichment amount of 2 g plant sterols per proposed food serving size results in an intake maximum of 13 g/d. PMID- 15877879 TI - Cold-induced heat production preceding shivering. AB - Individual changes in heat production and body temperature were studied in response to cold exposure, prior to shivering. The subjects ten women (seven men) were of normal weight, had a mean age of 23 (SD 3) years and average BMI 22.2 (SD 1.6) Kg/m2. They were lying supine under thermoneutral conditions for 30 min and were subsequently exposed to air of 15 degrees C until shivering occurred. Heat production was measured with a ventilated hood. Body composition was measured with underwater weighing and 2H dilution. Body temperatures were measured with thermistors. Heat production during cold exposure prior to shivering increased and reached a plateau. Skin temperature decreased and did not reach a plateau during the test period. The non-shivering interval (NSI) ranged from 20 to 148 min, was not related to body composition and was not significantly different between women (81 (sd 15) min) and men (84 (sd 34) min). NSI was negatively related to skin temperature (r2 0.44, P=0.004), and skin temperature was related to heat production (r2 0.39, P=0.007). In conclusion, subjects with a relatively large heat production during cold exposure maintained a relatively high skin temperature but showed a short NSI, independent of differences in body composition. PMID- 15877880 TI - Lignan contents of Dutch plant foods: a database including lariciresinol, pinoresinol, secoisolariciresinol and matairesinol. AB - Enterolignans (enterodiol and enterolactone) can potentially reduce the risk of certain cancers and cardiovascular diseases. Enterolignans are formed by the intestinal microflora after the consumption of plant lignans. Until recently, only secoisolariciresinol and matairesinol were considered enterolignan precursors, but now several new precursors have been identified, of which lariciresinol and pinoresinol have a high degree of conversion. Quantitative data on the contents in foods of these new enterolignan precursors are not available. Thus, the aim of this study was to compile a lignan database including all four major enterolignan precursors. Liquid chromatography-tandem mass spectrometry was used to quantify lariciresinol, pinoresinol, secoisolariciresinol and matairesinol in eighty-three solid foods and twenty-six beverages commonly consumed in The Netherlands. The richest source of lignans was flaxseed (301,129 microg/100 g), which contained mainly secoisolariciresinol. Also, lignan concentrations in sesame seeds (29,331 microg/100 g, mainly pinoresinol and lariciresinol) were relatively high. For grain products, which are known to be important sources of lignan, lignan concentrations ranged from 7 to 764 microg/100 g. However, many vegetables and fruits had similar concentrations, because of the contribution of lariciresinol and pinoresinol. Brassica vegetables contained unexpectedly high levels of lignans (185-2321 microg/100 g), mainly pinoresinol and lariciresinol. Lignan levels in beverages varied from 0 (cola) to 91 microg/100 ml (red wine). Only four of the 109 foods did not contain a measurable amount of lignans, and in most cases the amount of lariciresinol and pinoresinol was larger than that of secoisolariciresinol and matairesinol. Thus, available databases largely underestimate the amount of enterolignan precursors in foods. PMID- 15877881 TI - Meal-based intake assessment tool: relative validity when determining dietary intake of Fe and Zn and selected absorption modifiers in UK men. AB - A computer-based dietary assessment tool, the meal-based intake assessment tool (MBIAT), is described. In the current study, dietary intakes of Fe and Zn fractions (total Fe, non-haem Fe, haem Fe, meat Fe, total Zn) and dietary components that influence Fe and Zn absorption (vitamin C, phytate, Ca, grams of meat/fish/poultry, black tea equivalents, phytate:Zn molar ratio) were assessed. The relative validity of the MBIAT was determined in forty-eight UK men aged 40 years and over by comparing its results with those from weighed diet records collected over 12 d. There was good agreement between the MBIAT and the weighed diet records for median intakes of total, non-haem, haem and meat Fe, Zn, vitamin C, phytate, grams of meat/fish/poultry and phytate:Zn molar ratio. Correlations between the two methods ranged from 0.32 (for Ca) to 0.80 (for haem Fe), with 0.76 for total Fe and 0.75 for Zn. The percentage of participants classified by the MBIAT into the same/opposite weighed diet record quartiles ranged from 56/0 for Fe and 60/0 for Zn to 33/10 for Ca. The questionnaire also showed an acceptable level of agreement between repeat administrations (e.g. a correlation for total Fe of 0.74). In conclusion, the MBIAT is appropriate for assessing group dietary intakes of total Fe and Zn and their absorption modifiers in UK men aged 40 years and over. PMID- 15877882 TI - The concept of well-being: relevance to nutrition research. AB - The aim of this paper is to discuss issues that fall within the general concept of well-being, with special emphasis on approaches that have been used in studies of nutrition and behaviour. Following this, two specific studies are described in detail, the first examining high-fibre breakfast cereals and the second investigating effects of inulin. Studies of nutrition and well-being can be categorised in a number of ways. One method involves examining acute effects of nutrition on mood and cognitive functioning. Another method has been to examine cross-sectional associations between dietary habits and questionnaire measures of reported health. Examples are given showing that regular consumption of a high fibre diet is associated with better-reported physical and mental health. The problem with such correlational studies is that it is impossible to infer causality. Intervention studies are necessary to achieve this and some examples of this approach are given. In the first study reported here, we examined whether consumption of high-fibre breakfast cereal led to an increase in energy. Such an effect was observed and plausible biological mechanisms underlying such results are described. A similar methodology has recently been used to examine the effects of inulin. In this case the results showed no negative side-effects of taking inulin but there were no beneficial effects of inulin on measures of well being (both subjective reports and objective measures). Possible reasons for these effects are discussed. PMID- 15877883 TI - Methodological considerations in measuring human calcium absorption: relevance to study the effects of inulin-type fructans. AB - During the last 50 years, a variety of methods have been developed to estimate Ca absorption in man. Mass balances were initially used, but these were unable to accurately measure fractional Ca absorption because they cannot distinguish unabsorbed dietary Ca from endogenous faecal Ca excretion (excretion of previously absorbed Ca back into the gut). A number of isotopic methods have been developed that can measure true fractional Ca absorption, employing radioisotopes, stable isotopes, or both. Different methods involve collection of urine, faecal or plasma samples. Of the currently available methods, the dual isotope tracer method with a timed urine collection is probably the most precise and reliable. It is also relatively straightforward to carry out and avoids the need for a faecal collection. The purpose of the present paper is to discuss the general advantages and disadvantages of the different methods of Ca absorption. In addition, the limitations the different methods have in examining the possible effects of non-digestible oligosaccharides on Ca absorption will be discussed. PMID- 15877884 TI - Inulin-type fructans and bone health: state of the art and perspectives in the management of osteoporosis. AB - If the primary role of diet is to provide sufficient nutrients to meet the metabolic requirements of an individual, there is an emerging rationale to support the hypothesis that, by modulating specific target functions in the body, it can help achieve optimal health. Regarding osteoporosis prevention, since Ca is most likely to be inadequate in terms of dietary intake, every strategy targeting an improvement in Ca absorption is very interesting. Actually, this process may be susceptible to manipulation by fermentable substrates. In this light, inulin-type fructans are very interesting, even if we need to gather more data targeting bone metabolism before health professionals can actively advocate their consumption to prevent senile osteoporosis. Besides targeting the prevention of postmenopausal osteoporosis, inulin-type fructans still remain a source for putative innovative dietary health intervention. Indeed, given in combination with isoflavones, they may have a potential for maintaining or improving the bone mass of human subjects, by modulating the bioavailability of phyto-oestrogens. PMID- 15877885 TI - Application of inulin-type fructans in animal feed and pet food. AB - The inulin-type fructans are non-digestible oligosaccharides that are fermented in the gastrointestinal tract of farm animals and pets. This review focuses on the various effects of inulin-type fructans in pigs, poultry, calves and companion animals. Effects of the inulin-type fructans on gut microflora, digestion and availability of nutrients, gut morphology, fermentation characteristics and animal performance are discussed. Inulin-type fructans can support animal performance and health by affecting nutrient digestion, gut microflora and gut morphology, although results vary depending on composition of the basal diet, inclusion level, type of fructan, adaptation period and experimental hygienic conditions. PMID- 15877886 TI - Introducing inulin-type fructans. AB - Inulin is a generic term to cover all beta(2-->1) linear fructans. Chicory inulin is a linear beta(2-->1) fructan (degree of polymerisation (DP) 2 to 60; DPav=12), its partial enzymatic hydrolysis product is oligofructose (DP 2 to 8; DPav=4), and by applying specific separation technologies a long-chain inulin known as inulin HP (DP 10 to 60; DPav=25) can be produced. Finally, a specific product known as oligofructose-enriched inulin is obtained by combining chicory long chain inulin and oligofructose. Because of the beta-configuration of the anomeric C2 in their fructose monomers, inulin-type fructans resist hydrolysis by intestinal digestive enzymes, they classify as 'non-digestible' carbohydrates, and they are dietary fibres. By increasing faecal biomass and water content of the stools, they improve bowel habits, but they have characteristic features different from other fibres. They affect gastrointestinal functions not because of their physico-chemical properties but rather because of their biochemical and physiological attributes. In the colon, they are rapidly fermented to produce SCFA that are good candidates to explain some of the systemic effects of inulin type fructans. Fermentation of inulin-type fructans in the large bowel is a selective process; bifidobacteria (and possibly a few other genera) are preferentially stimulated to grow, thus causing significant changes in the composition of the gut microflora by increasing the number of potentially health promoting bacteria and reducing the number of potentially harmful species. Both oligofructose and inulin are prebiotic. They also induce changes in colonic epithelium stimulating proliferation in the crypts, increasing the concentration of polyamines, changing the profile of mucins, and modulating endocrine as well as immune functions. From a nutrition labelling perspective, inulin-type fructans are not only prebiotic dietary fibres; they are also low-calorie carbohydrates [6.3 kJ/g (1.5 kcal/g)]. Supported by the results of a large number of animal studies and human nutrition intervention trials, the claim 'inulin-type fructans enhance calcium and magnesium absorption' is scientifically substantiated, but different inulin-type fructans have probably a different efficacy (in terms of effective daily dose), the most active product being the oligofructose-enriched inulin. A series of animal studies demonstrate that inulin-type fructans affect the metabolism of lipids primarily by decreasing triglyceridaemia because of a reduction in the number of plasma VLDL particles. The human data largely confirm the animal experiments. They demonstrate mainly a reduction in triglyceridaemia and only a relatively slight decrease in cholesterolaemia mostly in (slightly) hypertriglyceridaemic conditions. Inulin appears thus eligible for an enhanced function claim related to normalization of blood triacylglycerols. A large number of animal data convincingly show that inulin-type fructans reduce the risk of colon carcinogenesis and nutrition intervention trials are now performed to test that hypothesis in human subjects known to be at risk for polyps and cancer development in the large bowel. PMID- 15877887 TI - Modelling dishes and exploring culinary 'precisions': the two issues of molecular gastronomy. AB - The scientific strategy of molecular gastronomy includes modelling 'culinary definitions' and experimental explorations of 'culinary precisions'. A formalism that describes complex dispersed systems leads to a physical classification of classical sauces, as well as to the invention of an infinite number of new dishes. PMID- 15877888 TI - The incretins: a link between nutrients and well-being. AB - The glucoincretins, glucagon-like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP), are intestinal peptides secreted in response to glucose or lipid intake. Data on isolated intestinal tissues, dietary treatments and knockout mice strongly suggest that GIP and GLP-1 secretion requires glucose and lipid metabolism by intestinal cells. However, incretin secretion can also be induced by non-digestible carbohydrates and involves the autonomic nervous system and endocrine factors such as GIP itself and cholecystokinin. The classical pharmacological approach and the recent use of knockout mice for the incretin receptors have shown that a remarkable feature of incretins is the ability to stimulate insulin secretion in the presence of hyperglycaemia only, hence avoiding any hypoglycaemic episode. This important role is the basis of ongoing clinical trials using GLP-1 analogues. Since most of the data concern GLP-1, we will focus on this incretin. In addition, GLP-1 is involved in glucose sensing by the autonomic nervous system of the hepato-portal vein controlling muscle glucose utilization and indirectly insulin secretion. GLP-1 has been shown to decrease glucagon secretion, food intake and gastric emptying, preventing excessive hyperglycaemia and overfeeding. Another remarkable feature of GLP-1 is its secretion by the brain. Recently, elegant data showed that cerebral GLP-1 is involved in cognition and memory. Experiments using knockout mice suggest that the lack of the GIP receptor prevents diet-induced obesity. Consequently, macronutrients controlling intestinal glucose and lipid metabolism would control incretin secretion and would consequently be beneficial for health. The control of incretin secretion represents a major goal for new therapeutic as well as nutrition strategies for treating and/or reducing the risk of hyperglycaemic syndromes, excessive body weight and thus improvement of well-being. PMID- 15877889 TI - Impact of inulin and oligofructose on gastrointestinal peptides. AB - In the present paper, we summarise the data supporting the following hypothesis: dietary inulin-type fructans extracted from chicory root may modulate the production of peptides, such as incretins, by endocrine cells present in the intestinal mucosa, this phenomenon being involved in the regulation of food intake and/or systemic effects. To test this hypothesis, male Wistar rats received for 3 weeks either a standard diet or the same diet supplemented with 10 % inulin-type fructans with different degrees of polymerisation. All the effects were most pronounced with the diet containing oligofructose, and consisted of (i) a decrease in mean daily energy intake and in epididymal fat mass; (ii) a higher caecal pool of the anorexigenic glucagon-like peptide-1 (7-36) amide (GLP-1), and peptide YY (PYY), due to caecal tissue proliferation; (iii) an increase in GLP-1 and of its precursor - proglucagon mRNA - concentrations in the proximal colon; (iv) an increase in portal serum level of GLP-1 and PYY; (v) a decrease in serum orexigenic peptide ghrelin. Moreover, oligofructose supplementation improved glucose homeostasis (i.e. decreased glycaemia, increased pancreatic and serum insulin content) in diabetic rats previously treated with streptozotocin, a phenomenon that is partly linked to the reduction in food intake and that correlates with the increase in colic and portal GLP-1 content. Based on these results it appears justified to test, in human subjects, the hypothesis that dietary inulin-type fructans could play a role in the management of obesity and diabetes through their capacity to promote secretion of endogenous gastrointestinal peptides involved in appetite regulation. PMID- 15877890 TI - Effects of inulin-type fructans on lipid metabolism in man and in animal models. AB - Studies in rodents show that inulin and oligofructose can reduce the plasma levels of cholesterol and triacylglycerols (TG). In addition, they can oppose TG accumulation in liver and have favourable effects on hepatic steatosis. The hypotriglyceridaemic effect is due to a reduction in hepatic re-esterification of fatty acids, but mainly in the expression and activity of liver lipogenesis, resulting in lower hepatic secretion rate of TG. This repression of lipogenesis is not observed in adipose tissue. The effect on liver lipogenesis can be explained by reduced insulin/glucose levels or by a selective exposure of the liver to increased amounts of propionic acid produced in the large intestine during fermentation of non-digestible carbohydrates. The decrease in plasma cholesterol could also be due to inhibition of cholesterol synthesis by propionic acid or to modifications in the bile acid metabolism. Studies in man yield more conflicting results with a decrease or no effects on plasma lipid levels, and, when a decrease is observed, more marked effects on TG than on cholesterol and more consistent action of inulin than of oligofructose. Besides the difference in the dose of inulin or oligofructose used, differences in metabolic status could play a role in this discrepancy between man and animals since reduction in plasma TG is observed in man mainly in a situation of increased liver lipogenesis (high carbohydrate diet, obesity, hypertriglyceridaemia). The effects on plasma cholesterol appear also more marked in hyperlipidaemic subjects than in healthy controls, suggesting that inulin and oligofructose have beneficial effects in these types of subjects. PMID- 15877891 TI - Behavioural and cognitive effects of oligofructose-enriched inulin in rats. AB - The behavioural and cognitive effects of oligofructose-enriched inulin at the doses of 5 and 10 % in the diet, orally ingested daily during 2 weeks, were investigated using a functional observational battery (FOB) and the light extinction test in male Wistar rats. Control rats received a standard diet and were tested in the same test situations. The behavioural effects were assessed 2 d before and 14 d after the beginning of the treatment period and the cognitive effects were investigated after the administration period by lever-pressing activity and learning discrimination using the light extinction test paradigm. In general, the study demonstrated that oligofructose-enriched inulin at 5 % in the diet, and particularly at 10 % in the diet, caused relaxing-like effects, stimulated and increased the general activity and interest of the rats to the test environment. In addition, both doses of oligofructose-enriched inulin showed significant effects on learning discrimination in male rats, in comparison with the control diet. These results suggest that oligofructose-enriched inulin, particularly at the dose of 10 %, improves cognitive performances in the light extinction test and the well-being of male rats using the FOB. PMID- 15877892 TI - Prebiotics and resistance to gastrointestinal infections. AB - Acute gut disorder is a cause for significant medicinal and economic concern. Certain individual pathogens of the gut, often transmitted in food or water, have the ability to cause severe discomfort. There is a need to manage such conditions more effectively. The route of reducing the risk of intestinal infections through diet remains largely unexplored. Antibiotics are effective at inhibiting pathogens; however, these should not be prescribed in the absence of disease and therefore cannot be used prophylactically. Moreover, their indiscriminate use has reduced effectiveness. Evidence has accumulated to suggest that some of the health-promoting bacteria in the gut (probiotics) can elicit a multiplicity of inhibitory effects against pathogens. Hence, an increase in their numbers should prove effective at repressing pathogen colonisation if/when infectious agents enter the gut. As such, fortification of indigenous bifidobacteria/lactobacilli by using prebiotics should improve protection. There are a number of potential mechanisms for lactic acid bacteria to reduce intestinal infections. Firstly, metabolic endproducts such as acids excreted by these micro-organisms may lower the gut pH to levels below those at which pathogens are able to effectively compete. Also, many lactobacilli and bifidobacteria species are able to excrete natural antibiotics, which can have a broad spectrum of activity. Other mechanisms include an improved immune stimulation, competition for nutrients and blocking of pathogen adhesion sites in the gut. Many intestinal pathogens like type 1 fimbriated Escherichia coli, salmonellae and campylobacters utilise oligosaccharide receptor sites in the gut. Once established, they can then cause gastroenteritis through invasive and/or toxin forming properties. One extrapolation of the prebiotic concept is to simulate such receptor sites in the gut lumen. Hence, the pathogen is 'decoyed' into not binding at the host mucosal interface. The combined effects of prebiotics upon the lactic acid flora and anti adhesive strategies may lead towards new dietary interventions against food safety agents. PMID- 15877893 TI - Modulation of gut mucosal biofilms. AB - Non-digestible inulin-type fructans, such as oligofructose and high-molecular weight inulin, have been shown to have the ability to alter the intestinal microbiota composition in such a way that members of the microbial community, generally considered as health-promoting, are stimulated. Bifidobacteria and lactobacilli are the most frequently targeted organisms. Less information exists on effects of inulin-type fructans on the composition, metabolism and health related significance of bacteria at or near the mucosa surface or in the mucus layer forming mucosa-associated biofilms. Using rats inoculated with a human faecal flora as an experimental model we have found that inulin-type fructans in the diet modulated the gut microbiota by stimulation of mucosa-associated bifidobacteria as well as by partial reduction of pathogenic Salmonella enterica subsp. enterica serovar Typhimurium and thereby benefit health. In addition to changes in mucosal biofilms, inulin-type fructans also induced changes in the colonic mucosa stimulating proliferation in the crypts, increasing the release of mucins, and altering the profile of mucin components in the goblet cells and epithelial mucus layer. These results indicate that inulin-type fructans may stabilise the gut mucosal barrier. Dietary supplementation with these prebiotics could offer a new approach to supporting the barrier function of the mucosa. PMID- 15877894 TI - The role of gut-associated lymphoid tissues and mucosal defence. AB - The newborn infant leaves a germ-free intrauterine environment to enter a contaminated extrauterine world and must have adequate intestinal defences to prevent the expression of clinical gastrointestinal disease states. Although the intestinal mucosal immune system is fully developed after a full-term birth, the actual protective function of the gut requires the microbial stimulation of initial bacterial colonization. Breast milk contains prebiotic oligosaccharides, like inulin-type fructans, which are not digested in the small intestine but enter the colon as intact large carbohydrates that are then fermented by the resident bacteria to produce SCFA. The nature of this fermentation and the consequent pH of the intestinal contents dictate proliferation of specific resident bacteria. For example, breast milk-fed infants with prebiotics present in breast milk produce an increased proliferation of bifidobacteria and lactobacilli (probiotics), whereas formula-fed infants produce more enterococci and enterobacteria. Probiotics, stimulated by prebiotic fermentation, are important to the development and sustainment of intestinal defences. For example, probiotics can stimulate the synthesis and secretion of polymeric IgA, the antibody that coats and protects mucosal surfaces against harmful bacterial invasion. In addition, appropriate colonization with probiotics helps to produce a balanced T helper cell response (Th1=Th2=Th3/Tr1) and prevent an imbalance (Th1>Th2 or Th2>Th1) contributing in part to clinical disease (Th2 imbalance contributes to atopic disease and Th1 imbalance contributes to Crohn's disease and Helicobacter pylori-induced gastritis). Furthermore, a series of pattern recognition receptors, toll-like receptors on gut lymphoid and epithelial cells that interact with bacterial molecular patterns (e.g. endotoxin (lipopolysaccharide), flagellin, etc.), help modulate intestinal innate immunity and an appropriate adaptive immune response. Animal and clinical studies have shown that inulin-type fructans will stimulate an increase in probiotics (commensal bacteria) and these bacteria have been shown to modulate the development and persistence of appropriate mucosal immune responses. However, additional studies are needed to show that prebiotics can directly or indirectly stimulate intestinal host defences. If this can be demonstrated, then prebiotics can be used as a dietary supplement to stimulate a balanced and an appropriately effective mucosal immune system in newborns and infants. PMID- 15877895 TI - Inulin, oligofructose and immunomodulation. AB - Diet is known to modulate immune functions in multiple ways and to affect host resistance to infections. Besides the essential nutrients, non-essential food constituents such as non-digestible carbohydrates may also have an impact on the immune system, especially in the area of the gut-associated lymphoid tissue (GALT). Recent data now provide first evidence that prebiotics such as inulin/oligofructose (IN/OF) modulate functions of the immune system. In animal studies IN/OF primarily activated immune cells in Peyer's patches including IL-10 production and natural killer (NK) cell cytotoxicity. Other immune functions modulated by IN/OF included the concentration of secretory IgA in ileum and caecum, splenic NK cell cytotoxicity as well as splenocyte cytokine production. In different tumour models, a lower incidence of tumours was observed, which in the case of colonic tumours was associated with enhanced NK cell cytotoxicity in the GALT. Few human studies so far have investigated the effects of IN/OF alone or in combination with other dietary supplements on immunocompetence. Supplementation of IN/OF resulted in minor changes of systemic immune functions such as decrease in phagocytic activity. No data are available on the effects of IN/OF on the GALT in man. The mechanisms of the reported effects of IN/OF on the immune system are currently investigated and include: (i) direct effects of lactic acid-producing bacteria or bacterial constituents on immune cells; (ii) the production of SCFA and binding to SCFA receptors on leucocytes. In conclusion, the current data suggest that IN/OF primarily modulate immune parameters in the GALT, but splenocytes are also activated by IN/OF. Human studies are needed to find out whether IN/OF have the potential to modulate systemic immunity in well-nourished individuals and to lower the risk of diseases such as colon cancer. PMID- 15877896 TI - Application of prebiotics in infant foods. AB - The rationale for supplementing an infant formula with prebiotics is to obtain a bifidogenic effect and the implied advantages of a 'breast-fed-like' flora. So far, the bifidogenic effect of oligofructose and inulin has been demonstrated in animals and in adults, of oligofructose in infants and toddlers and of a long chain inulin (10 %) and galactooligosaccharide (90 %) mixture in term and preterm infants. The addition of prebiotics to infant formula softens stools but other putative effects remain to be demonstrated. Studies published post marketing show that infants fed a long-chain inulin/galactooligosaccharide mixture (0.8 g/dl) in formula grow normally and have no side-effects. The addition of the same mixture at a concentration of 0.8 g/dl to infant formula was therefore recognized as safe by the European Commission in 2001 but follow-up studies were recommended. It is thought that a bifidogenic effect is beneficial for the infant host. The rising incidence in allergy during the first year of life may justify the attempts to modulate the infant's flora. Comfort issues should not be confused with morbidity and are likely to be multifactorial. The functional effects of prebiotics on infant health need further study in controlled intervention trials. PMID- 15877897 TI - Inulin and oligofructose: impact on intestinal diseases and disorders. AB - A large and diverse variety of bacteria have evolved and adapted to live in the human intestinal habitat in a symbiotic arrangement that influences both physiology and pathology in the host. Symbiosis between host and flora can be optimised by prebiotics. Inulin-type fructans have been shown to improve the metabolic functions of the commensal flora. Clinical and experimental data suggest that they also improve the gut mucosal barrier. Furthermore, modulation of the trophic functions of the flora by these prebiotics could help in the prevention of inflammatory bowel diseases. The anti-inflammatory effects of inulin or oligofructose have been assessed in the rat model of distal colitis induced by dextran sodium sulphate, which histologically resembles human ulcerative colitis, and in the trinitrobenzene sulphonic acid model that resembles human Crohn's disease. Both inulin and oligofructose stimulate colonic production of SCFA and favour the growth of indigenous lactobacilli and/or bifidobacteria. These effects are associated with reduced mucosal inflammation and decreased mucosal lesion scores. Inulin has also been tested in a placebo controlled clinical trial in patients with relapsing pouchitis. Treatment reduced endoscopic and histological parameters of inflammation of the pouch mucosa. Inulin and oligofructose may offer an opportunity to prevent chronic inflammatory intestinal disorders, and this potential should be tested in further clinical studies. PMID- 15877898 TI - Mucosal bacteria in ulcerative colitis. AB - Ulcerative colitis (UC) is an acute and chronic inflammatory bowel disease of unknown aetiology, although bacterial species belonging to the normal colonic microbiota are known to be involved in its initiation and maintenance. Several organisms have been linked to the disease; however, mucosa-associated bacteria are more likely to be involved than their luminal counterparts, due to their close proximity to the host epithelium. Comparative bacteriological analyses were done on rectal biopsies to investigate differences in mucosal bacteria in patients with UC and healthy controls. Complex bacterial communities were found in both groups, with significant reductions in bifidobacterial numbers in UC, which suggested that they might have a protective role in the disease. Accordingly, a therapy for treating UC was designed, with the aim of modifying the mucosal microbiota to increase bifidobacterial colonisation and reduce inflammation. Ranges of mucosal and faecal bifidobacteria were tested for their substrate preferences and their abilities to survive under a variety of environmental conditions. A synbiotic comprising a probiotic (Bifidobacterium longum) isolated from healthy rectal mucosa combined with a prebiotic (oligofructose-enriched inulin - Synergy 1) was developed. The treatment was used in a randomised controlled trial involving eighteen patients with active UC, for a period of 1 month. Rectal biopsies were collected at the beginning and end of the study. Bacteriological analysis and transcription levels of epithelium related immune markers were assessed. Results demonstrated that short-term synbiotic treatment resulted in increased bifidobacterial colonisation of the rectal mucosa and induced significant reductions in the expression of molecules that control inflammation in active UC. PMID- 15877899 TI - Intestinal perception: mechanisms and assessment. AB - Physiological stimuli in the gut induce regulatory reflexes to accomplish the digestive process, but are normally not perceived. However, under some circumstances, gut stimuli may activate perception pathways and induce conscious sensations. Experimental evidence gathered during the past decade suggests that patients with functional gut disorders and unexplained abdominal symptoms may have a sensory dysfunction of the gut, so that physiological stimuli would induce symptoms. Assessment of visceral sensitivity is still poorly developed, but in analogy to somatosensory testing, differential stimulation of visceral afferents may be achieved by a combination of stimulation techniques, which may help to characterize sensory dysfunctions. Visceral afferent input is modulated by a series of mechanisms at different levels of the brain-gut axis, and conceivably, a dysfunction of these regulatory mechanisms could cause hyperalgesia. The sensory dysfunction in functional patients seems to be associated with altered reflex activity, and both mechanisms may interact to produce the symptoms. Evidence of a gut sensory-reflex dysfunction as a common pathophysiological mechanism in different functional gastrointestinal disorders would suggest that they are different forms of the same process, and that the clinical manifestations depend on the specific pathways affected. PMID- 15877900 TI - Inulin-type fructans and reduction in colon cancer risk: review of experimental and human data. AB - Inulin-type fructans (beta(2,1)fructans) extracted from chicory roots (Cichorium intybus) are prebiotic food ingredients, which in the gut lumen are fermented to lactic acid and SCFA. Research in experimental animal models revealed that inulin type fructans have anticarcinogenic properties. A number of studies report the effects of inulin-type fructans on chemically induced pre-neoplastic lesions (ACF) or tumours in the colon of rats and mice. In twelve studies, there were twenty-nine individual treatment groups of which twenty-four measured aberrant crypt foci (ACF) and five measured tumours. There was a significant reduction of ACF in twenty-one of the twenty-four treatment groups and of tumour incidence in five of the five treatment groups. Higher beneficial effects were achieved by synbiotics (mixtures of probiotics and prebiotics), long-chain inulin-type fructans compared to short-chain derivatives, and feeding high-fat Western style diets. Inulin-type fructans reduced tumour incidence in APC(Min) mice in two of four studies and reduced growth and metastasising properties of implanted tumour cells in mice (four studies). The effects have been reported to be associated with gut flora-mediated fermentation and production of butyrate. In human cells, inulin-derived fermentation products inhibited cell growth, modulated differentiation and reduced metastasis activities. In conclusion, evidence has been accumulated that shows that inulin-type fructans and corresponding fermentation products reduced the risks for colon cancer. The involved mechanisms included the reduction of exposure to risk factors and suppression of tumour cell survival. Thus, this specific type of dietary fibre exerted both blocking agent and suppressing agent types of chemopreventive activities. PMID- 15877901 TI - The SYNCAN project: goals, set-up, first results and settings of the human intervention study. AB - Experimental evidence on the anticancer properties of dietary prebiotics such as chicory inulin and oligofructose and dietary probiotics has accumulated in recent years. Various experimental models ranging from chemoprevention studies, tumour implantation models to genetically modified mice models, etc. have systematically shown the protective effects of these food ingredients. In some studies it appeared that synbiotics (combination of pre- and probiotics) exerted synergistic activity against processes of carcinogenesis. The logical next step in research was to find out if these observations also would be valid for human volunteers. This was the principal goal of the EU-sponsored SYNCAN project (QLK1-1999-346) which involved the integration of an in vitro study to select the most suitable synbiotic preparation, the application of this synbiotic in an in vivo rat model of chemically induced colon cancer, and, as the heart of the project, the investigation of the synbiotic effects in a human intervention study. The in vitro tests consisted of fermentation studies where the interaction of pre- and probiotics was studied. Cell-free supernatants were generated from various synbiotic combinations fermented by faecal slurry, which were then used to optimise a series of bioassays. In the rat study the anticarcinogenic effect of prebiotics and synbiotics but not of probiotics was demonstrated. Using tissue samples generated in this model, attempts were made to gain a better insight into the mechanisms underlying cancer development. The human intervention study consisted of two groups of volunteers. One group was composed of people at high risk (polypectomised subjects) for colon cancer and the other of volunteers (colon cancer subjects) who had previously undergone 'curative resection' for colon cancer but were not currently receiving treatment. The present paper describes the experimental design of the SYNCAN study, and demonstrates a functional effect of the synbiotic preparation (probiotic survival during gastrointestinal transit and modification of the intestinal flora). Detailed experimental outcome of the human intervention study will be reported elsewhere. PMID- 15877902 TI - Inulin, oligofructose and bone health: experimental approaches and mechanisms. AB - Inulin-type fructans have been proposed to benefit mineral retention, thereby enhancing bone health. Many, but not all, experimental animal studies have shown increased mineral absorption by feeding non-digestible oligosaccharides. Possible reasons for inconsistencies are explored. A few studies have reported an enhanced bone mineral density or content. Bone health can be evaluated in chronic feeding studies with bone densitometry, bone breaking strength, bone mineral concentration and bone structure. Isotopic Ca tracers can be used to determine the point of metabolism affected by feeding a functional food ingredient. These methods and the effects of feeding inulin-type fructose are reviewed. Inulin-type fructans enhance Mg retention. Chicory long-chain inulin and oligofructose enhance femoral Ca content, bone mineral density and Ca retention through enhanced Ca absorption and suppressed bone turnover rates, but it is not bone promoting under all conditions. PMID- 15877903 TI - Public health nutrition. PMID- 15877904 TI - Out of the box. PMID- 15877905 TI - Effect of iron supplementation on mental and motor development in children: systematic review of randomised controlled trials. AB - OBJECTIVE: To evaluate the effect of iron supplementation on mental and motor development in children through a systematic review of randomised controlled trials (RCTs). DATA SOURCES: Electronic databases, personal files, hand search of reviews, bibliographies of books, abstracts and proceedings of international conferences. REVIEW METHODS: RCTs with interventions that included oral or parenteral iron supplementation, fortified formula milk or cereals were evaluated. The outcomes studied were mental and motor development scores and various individual development tests employed, including Bayley mental and psychomotor development indices and intelligence quotient. RESULTS: The pooled estimate (random effects model) of mental development score standardised mean difference (SMD) was 0.30 (95% confidence interval (CI) 0.15 to 0.46, P<0.001; P<0.001 for heterogeneity). Initial anaemia and iron-deficiency anaemia were significant explanatory variables for heterogeneity. The pooled estimate of Bayley Mental Development Index (weighted mean difference) in younger children (<27 months old) was 0.95 (95% CI -0.56 to 2.46, P=0.22; P=0.016 for heterogeneity). For intelligence quotient scores (> or =8 years age), the pooled SMD was 0.41 (95% CI 0.20 to 0.62, P<0.001; P=0.07 for heterogeneity). There was no effect of iron supplementation on motor development score (SMD 0.09, 95% CI 0.08 to 0.26, P=0.28; P=0.028 for heterogeneity). CONCLUSIONS: Iron supplementation improves mental development score modestly. This effect is particularly apparent for intelligence tests above 7 years of age and in initially anaemic or iron-deficient anaemic subjects. There is no convincing evidence that iron treatment has an effect on mental development in children below 27 months of age or on motor development. PMID- 15877906 TI - The CATCH Kids Club: a pilot after-school study for improving elementary students' nutrition and physical activity. AB - BACKGROUND: Although many school-based diet and physical activity interventions have been designed and evaluated, relatively few have been tested for the after school setting. After-school day-care programmes at either elementary schools or private locations provide a ready-made opportunity for health programmes that may be difficult to incorporate into an already-full school day. The purpose of this paper is to report on a pilot study of an after-school adaptation of the CATCH (Coordinated Approach To Child Health) elementary school programme called the CATCH Kids Club (CKC). METHODS: The CKC was pilot-tested and formatively evaluated in 16 Texas after-school programmes: eight in El Paso and eight in Austin (four intervention and four reference sites each). Evaluation consisted of direct observation of moderate to vigorous physical activity during play time, self-reported food intake and physical activity, and focus group interviews with after-school programme staff. RESULTS: Students responded well to the physical activity and snack components and were less interested in the five-module education component. Routine staff training was a key variable in achieving proper implementation; the ideal would be a full day with repeated follow-up model teaching visits. Staff turnover was a logistic issue, as was programme leader readiness and interest in conducting the programme. Strong and significant effects were observed for the physical activity but not for the education component. The results of the physical education component suggest it is feasible, effective and ready for larger-scale evaluation or dissemination. PMID- 15877907 TI - Overweight in urban, low-income, African American and Hispanic children attending Los Angeles elementary schools: research stimulating action. AB - OBJECTIVE: This study was undertaken to establish the prevalence and severity of nutritional problems among low-income children of elementary school age in the Los Angeles Unified School District (LAUSD) in order to collect baseline data to inform policy-makers. DESIGN AND METHODS: A cross-sectional survey of children in 14 elementary schools was conducted from January to June, 1998. Nine hundred and nineteen children were measured and interviewed. The planning, design and data analysis were carried out in collaboration with key LAUSD policy-makers. RESULTS: More than 35% of the sample was classified as being at risk for overweight or overweight according to body mass index. CONCLUSION: There is a high prevalence of children who are at risk for overweight or who are overweight in Los Angeles. This finding has triggered the development of multiple school-based intervention programmes. PMID- 15877908 TI - The stability of soft drinks intake from adolescence to adult age and the association between long-term consumption of soft drinks and lifestyle factors and body weight. AB - OBJECTIVES: To investigate the tracking of sugar-sweetened, carbonated soft drinks intake from age 15 to 33 years and the association between this intake and lifestyle factors and body weight. DESIGN: A longitudinal study with 18-20 years of follow-up. Data about diet, physical activity, smoking and dieting were collected in 1981/1979, 1991 and 1999. Body weight and height were measured in 1981/1979 and self-reported in 1999. SETTING: Oslo, Norway. SUBJECTS: Four hundred and twenty-two men and women. RESULTS: Tracking of soft drinks intake from adolescence into early adulthood (age 25 years) and from early adulthood into later adulthood (33 years) was moderate to high, while tracking from adolescence into later adulthood was low. Comparing those reporting a high intake of soft drinks in both 1991 and 1999 with those reporting a low intake at both times, male long-term high consumers were more likely to smoke (48 vs. 21%, P=0.002) and reported higher intakes of energy (12.2 vs. 10.2 MJ day(-1), P=0.005) and sugar (142 vs. 50 g day(-1), P<0.001) in 1999 than did long-term low consumers. Women high consumers were less likely to be physically active (14 vs. 42%, P=0.03) and had higher sugar intake (87 vs. 41 g day(-1), P<0.001) in 1999 than did women low consumers. There were no differences in body mass index, overweight or obesity in 1999 between long-term high and low consumers. CONCLUSION: In this study, stability of soft drinks intake from age 15 to 25 years and from age 25 to 33 years was moderate to high, while from age 15 to 33 years it was low. Soft drinks intake from age 25 to 33 years was associated with smoking and physical inactivity, but not with body weight. PMID- 15877909 TI - Who eats four or more servings of fruit and vegetables per day? Multivariate classification tree analysis of data from the 1998 Survey of Lifestyle, Attitudes and Nutrition in the Republic of Ireland. AB - OBJECTIVE: To identify, using the novel application of multivariate classification trees, the socio-economic, sociodemographic and health-related lifestyle behaviour profile of adults who comply with the recommended 4 or more servings per day of fruit and vegetables. DESIGN: Cross-sectional 1998 Survey of Lifestyle, Attitudes and Nutrition. SETTING: Community-dwelling adults aged 18 years and over on the Republic of Ireland electoral register. SUBJECTS: Six thousand five hundred and thirty-nine (response rate 62%) adults responded to a self-administered postal questionnaire, including a semi-quantitative food frequency questionnaire. RESULTS: The most important determining factor of compliance with the fruit and vegetable dietary recommendations was gender. A complex constellation of sociodemographic and socio-economic factors emerged for males whereas the important predictors of 4 or more servings of fruit and vegetable consumption among females were strongly socio-economic in nature. A separate algorithm was run to investigate the importance of health-related lifestyle and other dietary factors on compliance with the fruit and vegetable recommendations. Following an initial split on compliance with dairy recommendations, a combination of non-dietary behaviours showed a consistent pattern of healthier options more likely to lead to compliance with fruit and vegetable recommendations. There did, however, appear to be a compensatory element between the variables, particularly around smoking, suggesting the non existence of an exclusive lifestyle for health risk. CONCLUSIONS: Material and structural influences matter very much for females in respect to compliance with fruit and vegetable recommendations. For males, while these factors are important they appear to be mediated through other more socially contextual-type factors. Recognition of the role that each of these factors plays in influencing dietary habits of men and women has implications for the manner in which dietary strategies and policies are developed and implemented. PMID- 15877910 TI - Intake and sources of phylloquinone (vitamin K(1)) in 4-year-old British children: comparison between 1950 and the 1990s. AB - OBJECTIVES: To compare dietary intake and sources of phylloquinone (vitamin K1) in 4-year-old British children between 1950 and the 1990s, and report their variation by sociodemographic factors. DESIGN: Nationally representative samples of 4-year-olds from the longitudinal Medical Research Council National Survey of Health and Development (NSHD) (1950) and the cross-sectional National Diet and Nutrition Surveys (NDNS, 1992/93 and 1997). SETTING: Great Britain. SUBJECTS: Subjects were 4599 children born on 3-9 March 1946 (NSHD) and 307 children in the 1990s (NDNS). RESULTS: Geometric mean dietary phylloquinone intake was significantly higher in 1950 (39 mug day(-1), 95% confidence interval (CI) 37, 40) compared with the 1990s (24 mug day(-1), 95% CI 22, 25) (P<0.001). This difference remained when intake was expressed per MJ energy intake and per kilogram body weight, and after accounting for sex, region and occupational social class of the family. In 1950, phylloquinone intake in Scotland was significantly lower than in the rest of Britain. By the 1990s these regional differences had disappeared. Food sources of phylloquinone intake changed significantly between 1950 and the 1990s, with fats and oils contributing more and vegetables less, although vegetables contributed most (60% and 48%, respectively) to phylloquinone intake in both surveys. CONCLUSIONS: Phylloquinone intakes of children have decreased significantly since 1950. With the suggested need for adequate phylloquinone intake for optimal development and maintenance of bone and the cardiovascular system, the substantially lower phylloquinone intakes reported in children of the 1990s, compared with 1950, may have implications for the health of these two systems in later adulthood. PMID- 15877911 TI - Development and simulated validation of a food-frequency questionnaire for the Colombian population. AB - OBJECTIVE: To develop a food-frequency questionnaire (FFQ) useful for ranking of nutrient intakes. DESIGN: Subjects consuming their regular diet completed 7 days of weighed intake registry (7-WIR). Foods for the FFQ were selected by stepwise multiple regression. The FFQ was then completed for each subject using data on individual food consumption from the 7-WIR. The correlation and agreement between the extrapolated FFQ and the 7-WIR data were assessed using Spearman's rank correlation coefficients (rS) and Bland and Altman's limits of agreement (LOA). SETTING: Bucaramanga, Colombia. SUBJECTS: We studied 97 randomly selected 20-40 year-old subjects. RESULTS: Sixty foods were selected for the FFQ. The 7-WIR and the extrapolated FFQ intake estimates correlated well. rS was 0.58 for energy, 0.53 for carbohydrate, 0.50 for total fat, and 0.48 for protein. For micronutrients, rS varied from 0.46 (manganese) to 0.71 (vitamin B12). FFQ average intake estimates were 83%, 80%, 86.2% and 86.4% of 7-WIR estimates for energy, carbohydrate, total fat and protein, respectively. LOA for these nutrients ranged between 45% and 165%. FFQ micronutrient intakes were on average 96% (median) of those from the 7-WIR, and the median lower and upper LOA were 50% and 203%. However, there was no indication that the degree of agreement varied with the level of intake. CONCLUSIONS: According to our simulated validation, this FFQ may be useful to rank subjects by nutrient intake. Its validity against standard independent measurements and its applicability to other subsets of the Colombian population should be carefully considered. PMID- 15877912 TI - Reliability and validity of a questionnaire to measure personal, social and environmental correlates of fruit and vegetable intake in 10-11-year-old children in five European countries. AB - OBJECTIVES: To investigate the internal consistency of the scales and the test retest reliability and predictive validity of behaviour theory-based constructs measuring personal, social and environmental correlates of fruit and vegetable intake in 10-11-year-old children. DESIGN: Test-retest with one-week interval. SETTING: Five European countries: Norway, Spain, Denmark, Portugal, Belgium. SUBJECTS: Three hundred and twenty-six children completed the questionnaire during class hours. RESULTS: For the total sample across all countries, the test retest reliability was good to very good (intra-class correlation coefficient (ICC) >0.60) for 12 out of the 15 fruit constructs and also for 12 out of the 15 vegetable constructs. Acceptable ICCs, ranging between 0.50 and 0.59, were found for the remaining constructs. Test-retest reliability was comparable across countries. Only in Portugal were some significantly lower ICCs found for some constructs (knowledge and barriers related to fruit, general self-efficacy related to fruit and vegetables) compared with the other countries. Cronbach's alpha values were moderate to high (range 0.52 to 0.89) with the exception of the general self-efficacy scale, which had a value below 0.50 for both fruit (alpha=0.42) and vegetables (alpha=0.49). Spearman correlations with intake ranged between -0.16 and 0.54 for personal determinants and between 0.05 and 0.38 for environmental determinants. Compared with other studies, predictive validity can be considered moderate to good. CONCLUSIONS: The questionnaire provides a reliable, valid and easy-to-administer tool for assessing personal, social and environmental factors of potential influence on fruit and vegetable intake in 10 11-year-olds. PMID- 15877913 TI - Reproducibility of a short semi-quantitative food group questionnaire and its performance in estimating nutrient intake compared with a 7-day diet diary in the Million Women Study. AB - OBJECTIVES: To assess the short- and long-term reproducibility of a short food group questionnaire, and to compare its performance for estimating nutrient intakes in comparison with a 7-day diet diary. DESIGN: Participants for the reproducibility study completed the food group questionnaire at two time points, up to 2 years apart. Participants for the performance study completed both the food group questionnaire and a 7-day diet diary a few months apart. Reproducibility was assessed by kappa statistics and percentage change between the two questionnaires; performance was assessed by kappa statistics, rank correlations and percentages of participants classified into the same and opposite thirds of intake. SETTING: A random sample of participants in the Million Women Study, a population-based prospective study in the UK. SUBJECTS: In total, 12 221 women aged 50-64 years. RESULTS: In the reproducibility study, 75% of the food group items showed at least moderate agreement for all four time point comparisons. Items showing fair agreement or worse tended to be those where few respondents reported eating them more than once a week, those consumed in small amounts and those relating to types of fat consumed. Compared with the diet diary, the food group questionnaire showed consistently reasonable performance for the nutrients carbohydrate, saturated fat, cholesterol, total sugars, alcohol, fibre, calcium, riboflavin, folate and vitamin C. CONCLUSIONS: The short food group questionnaire used in this study has been shown to be reproducible over time and to perform reasonably well for the assessment of a number of dietary nutrients. PMID- 15877914 TI - A two-box option. PMID- 15877916 TI - Cost-effective measures to prevent obesity: epidemiological basis and appropriate target groups. AB - Cost-effective prevention strategies to prevent weight gain and the development of obesity should be based on appropriate knowledge of the determinants of weight gain. The body of evidence on the dietary determinants of weight gain is, however, fragmentary at best, partly because inappropriate research methods are used to study the determinants of obesity under normal circumstances. Evidence from studies using experimental diets have shortcomings because of their short duration and selection of highly-motivated subjects and because the outcomes can be easily influenced by the choice of foods to be used in the intervention. Of the observational studies, many have severe methodological shortcomings, e.g. ecological studies, cross-sectional surveys and classical cohort studies in which the baseline diet is linked to subsequent weight development over long periods of time. Longitudinal studies with repeated measurements in which changes in diet and physical activity are linked to changes in weight are probably the most informative, but these studies are relatively rare. There is a great interest in interventions that are effective and efficient for the prevention of obesity. Many countries and research funding agencies seem to show a strong tendency to develop interventions for children and adolescents exclusively. It can be easily shown, however, that intervention programmes are much more likely to be cost effective in older adults than in children, which indicates that adults should not be neglected as target populations for obesity prevention. Obesity prevention should follow a life-course approach, as currently recommended for non communicable diseases in general by the WHO. PMID- 15877917 TI - Obesity and metabolic disease: is adipose tissue the culprit? AB - Obesity is a risk factor for the development of type 2 diabetes and CVD. Is adipose tissue the culprit in the relationship between obesity and metabolic disease? It is certainly possible to argue that adipose tissue function is disturbed in obesity in such a way that adverse consequences may follow. For instance, lipolysis is down regulated, the sensitivity of lipolysis to insulin is reduced and there are disturbances in the regulation of adipose tissue blood flow. However, when examined critically these changes can be seen as adaptations to the increased adipose tissue mass, making the situation better rather than worse. In terms of the many peptide and other factors now known to be secreted from adipose tissue, it is easier to argue that adipose tissue is the culprit. However, for no single 'adipokine' is there as yet unequivocal evidence of a link between adipose tissue secretion and adverse metabolic events in other tissues. The best documented of these adipokines in relation to insulin resistance is adiponectin. Here, unusually, adiponectin confers insulin sensitivity, and its secretion is down regulated in obesity. It could be again that adipose tissue has down regulated its function in an attempt to compensate for its increased mass, although certainly that down-regulation is too extreme. On balance, it is clear that adipose tissue is a link in the chain of events leading to metabolic disease, but in many respects it is an innocent intermediary trying to deal with the consequences of positive energy balance, the real culprit. PMID- 15877918 TI - Endothelial dysfunction: role in obesity-related disorders and the early origins of CVD. AB - Atherosclerotic CVD is the most common cause of death in the West. Yet, its pathogenesis and early development are only partially understood. Central to the early atherosclerotic process is impairment of vascular endothelial function. Endothelial dysfunction can be measured non-invasively and is evident in children before clinical manifestations of atherosclerosis in adulthood. Factors in early life, such as conventional cardiovascular risk factors, or programming by perinatal growth and nutrition strongly affect endothelial function and hence the development of atherosclerosis and CVD. For instance, low birth weight and faster growth early in infancy have a detrimental effect on vascular structure and function. Childhood obesity, a key independent risk factor for CVD, also adversely affects early vascular health. Obesity is associated with endothelial dysfunction and greater arterial stiffness from as early as the first decade of life, while weight loss is beneficial. This effect on vascular function is probably mediated in part by low-grade inflammation and insulin resistance associated with obesity or by the production by adipose tissue of cytokine-like molecules, collectively termed adipokines. A high leptin concentration, in particular, is found in obese individuals and is strongly associated with vascular changes related to early atherosclerosis. The present review focuses on the early origins of endothelial dysfunction, emphasising the role of obesity. It also considers the mechanisms by which obesity impairs endothelial function, understanding of which will be important to further scientific knowledge and to improve public health. PMID- 15877919 TI - Fatty acids and the metabolic syndrome. AB - The metabolic syndrome is a very common condition, characterised by insulin resistance, dyslipidaemia, abdominal obesity and hypertension, that is associated with a high risk of type 2 diabetes mellitus (T2DM) and CVD. Obesity is a key aetiological factor in the development of the metabolic syndrome. In light of the increasing prevalence of obesity, there is a high requirement to reduce the impact of the adverse health effects associated with the metabolic syndrome. The aetiological role of nutrient-derived metabolic stressors, in particular fatty acids, in the development of obesity and the metabolic syndrome is explored. Also, the evidence that pro-inflammatory stressors may predispose to obesity induced insulin resistance is reviewed. The present paper explores the concept that reducing the impact of metabolic and inflammatory stressors may reduce the adverse health effects of obesity and slow the progression towards the metabolic syndrome and T2DM. Evidence from human dietary intervention studies that have investigated the potential therapeutic effects of dietary fatty acid modification is explored. The present review highlights the requirement to take account of genetic background, within the context of nutrient regulation of gene expression and individual responsiveness to dietary therapy. This approach will further the understanding of the interaction between fatty acids in the pathogenesis and progression of the metabolic syndrome. PMID- 15877920 TI - The biology of obesity. AB - Obesity is a multidisciplinary area, the 'biology' of which encompasses: (1) the fundamental mechanisms of energy balance and its regulation; (2) the biological basis for the development of obesity; (3) adipose tissue function; (4) the biological description of the obese state; (5) the pathological consequences of obesity; (6) the physiological basis for treatment strategies. At a mechanistic level, important developments in recent years include the identification of novel neuroendocrine factors in the control of appetite (such as cocaine- and amphetamine-regulated transcript, the orexins, the endocannabinoids) and the discovery of new peripheral signals (such as leptin, ghrelin). Despite the identification of additional uncoupling proteins (UCP2, UCP3), mitochondrial uncoupling in brown adipose tissue through UCP1 remains the only major mechanism for adaptive thermogenesis. White adipose tissue (WAT) has now moved centre stage in energy balance and obesity research, and there are three main reasons: (1) it is the organ which defines obesity; (2) it is the source of a critical endocrine signal in the control of body weight; (3) it secretes a range of diverse protein factors, termed adipokines, some of which are directly implicated in the pathologies associated with obesity. WAT is now recognised as a key endocrine organ, communicating both with the brain and peripheral tissues through the adipokines. Obesity is characterised by mild inflammation, and WAT may be the main locus of the inflammatory state, producing cytokines, chemokines, acute phase proteins and angiogenic factors. It has been suggested that inflammation in obesity is principally an adaptive response to hypoxia in clusters of adipocytes within the expanding adipose mass. PMID- 15877921 TI - Central regulation of energy balance: inputs, outputs and leptin resistance. AB - The regulation of energy balance is complex and, in man, imprecise. Nevertheless, in many individuals intake and expenditure are balanced with <1% error with little or no conscious effect. Essential components of such a regulatory system are signals, leptin and insulin, that reflect the size of lipid stores. Leptin receptors signal via phosphatidylinositol 3-kinase (as do insulin receptors) and via the transcription factor signal transducer and activator of transcription-3 to activate various types of neurone. Obese rodents, and possibly man, are resistant to leptin; in some cases because of genetic or perinatal programming (primary resistance), but commonly in response to high leptin levels (secondary resistance). Secondary leptin resistance may be a result of reduced transport of leptin to the brain or down-regulation of leptin signalling. Signals that reflect lipid stores form the tonic homeostatic regulatory system. They interact with episodic homeostatic signals carried by neurones, hormones and metabolites to regulate meal size and frequency. They also interact with signals related to the palatability of food, biorhythms and learning. Many neurotransmitters and hormones mediate responses to more than one input (e.g. gastric and adipocyte leptin), but are nevertheless most involved with particular inputs (e.g. leptin with adipocyte fat stores). Feeding can be divided into appetitive (preparation for feeding) and consummatory phases, which can both be further subdivided. Different sets of neurotransmitters and hormones are involved at each stage. In the long term it may be possible to customise obesity therapies according to those inputs and outputs that are most disturbed and most amenable to intervention in individual subjects. PMID- 15877922 TI - Respiration uncoupling and metabolism in the control of energy expenditure. AB - Metabolic energy expenditure negatively regulates energy balance. Metabolic and catabolic pathways contribute to energy expenditure. Catabolic pathways split C containing molecules into small molecules and generate reduced coenzymes and ATP. For a given amount of substrate, any increase in energy expenditure requires either increased ATP hydrolysis or decreased ATP synthesis. In skeletal muscles substrate utilisation is coupled to ATP production, whereas ATP hydrolysis is activated during physical exercise and increases energy expenditure. In brown adipose tissue activation of cells during exposure to cold increases substrate utilisation in such a way that glucose and fatty acid oxidation detach from the orthodox coupling to ATP synthesis and result in thermogenesis. The unique mechanism of uncoupling respiration that occurs in brown adipocyte mitochondria represents an attractive strategy for promoting energy expenditure and decreasing the fat content of the body. Moreover, ectopic expression of brown fat uncoupling protein (UCP) 1 in mouse skeletal muscle and induction of UCP1 in mouse or human white adipocytes promote fatty acid oxidation and resistance to obesity. In normal conditions UCP2 and UCP3 do not seem to contribute substantially to energy expenditure. Whether the induction of UCP1, the induction of other UCP or chemical mild uncoupling represent promising strategies for attenuating nutrient efficiency and counteracting obesity should be considered. PMID- 15877923 TI - Brain-adipose tissue cross talk. AB - While investigating the reversible seasonal obesity of Siberian hamsters, direct sympathetic nervous system (SNS) postganglionic innervation of white adipose tissue (WAT) has been demonstrated using anterograde and retrograde tract tracers. The primary function of this innervation is lipid mobilization. The brain SNS outflow to WAT has been defined using the pseudorabies virus (PRV), a retrograde transneuronal tract tracer. These PRV-labelled SNS outflow neurons are extensively co-localized with melanocortin-4 receptor mRNA, which, combined with functional data, suggests their involvement in lipolysis. The SNS innervation of WAT also regulates fat cell number, as noradrenaline inhibits and WAT denervation stimulates fat cell proliferation in vitro and in vivo respectively. The sensory innervation of WAT has been demonstrated by retrograde tract tracing, electrophysiological recording and labelling of the sensory-associated neuropeptide calcitonin gene-related peptide in WAT. Local injections of the sensory nerve neurotoxin capsaicin into WAT selectively destroy this innervation. Just as surgical removal of WAT pads triggers compensatory increases in lipid accretion by non-excised WAT depots, capsaicin-induced sensory denervation triggers increases in lipid accretion of non-capsaicin-injected WAT depots, suggesting that these nerves convey information about body fat levels to the brain. Finally, parasympathetic nervous system innervation of WAT has been suggested, but the recent finding of no WAT immunoreactivity for the possible parasympathetic marker vesicular acetylcholine transporter (VAChT) argues against this claim. Collectively, these data suggest several roles for efferent and afferent neural innervation of WAT in body fat regulation. PMID- 15877924 TI - Clinical treatment of obesity: are drugs and surgery the answer? AB - Obesity treatment remains a 'Cinderella' of all clinical management programmes, but generally without a happy ending. The great expectation for new therapeutic agents has not been fulfilled in clinical practice, whilst the restriction of eating through surgical division of the upper bowel seems strange in an age of advanced and sophisticated technology. The better understanding of the neuro regulation of appetite, and its application as part of evidence-based clinical interventions, could lead to a more coherent approach to obesity treatment. Nevertheless, investigation of potential neuroendocrine targets for appetite suppression suggests redundancy in the systems, which make development of effective agents against single receptors impractical. Importantly, the progressive rise in the prevalence of obesity will inevitably mean that only a small proportion of afflicted patients will actually be treated by long-term drugs and surgery. Drugs and surgery are not currently the answer for the majority of obese individuals. What is required is a better way of identifying patients who may particularly benefit from such approaches. However, the major emphasis must be the development of effective population-wide interventions that halt the increase in fatness and ensure that future generations maintain and enjoy a healthy body weight. PMID- 15877925 TI - Understanding the aetiology of childhood obesity: implications for treatment. AB - Childhood obesity poses one of the greatest challenges to paediatric health in the 21st century. Developing effective strategies for treatment and prevention is therefore a priority for clinical medicine and public health. This process is taking place at a time of unprecedented change in the understanding of the role of genetic factors in human health and disease, and genetic research into obesity has challenged assumptions about causal processes. In the spirit of the conference theme, the present paper will discuss the implications of the understanding of genetic and environmental influences on obesity for the development of effective behavioural treatments. PMID- 15877926 TI - Lifestyle strategies for weight control: experience from the Finnish Diabetes Prevention Study. AB - Currently, in many European countries more than half the adult population is overweight; it hass become 'abnormal' to be of 'normal weight'. The risk of type 2 diabetes, CVD, hypertension and certain forms of cancer increase with increasing weight. Biological evolution has produced body-fat-regulating mechanisms that are more powerful in protecting against weight loss than against weight gain. The current environment offers constant availability of affordable palatable energy-rich foods, with no need to consume the energy through physical activity. The 'obesogenic' environment is to some extent a political issue, but it has been shown that the healthcare system can also have a role in preventing obesity-related morbidity. The Finnish Diabetes Prevention Study was the first controlled randomised study to show that individualised lifestyle counselling of individuals with high risk of developing type 2 diabetes can influence diet, physical activity and body weight, and that type 2 diabetes can be prevented, or at least postponed. Most importantly, lifestyle changes do not have to be extreme. If the population would adopt a lifestyle in line with the official nutrition recommendations, the obesity and diabetes trend could at least be stabilised. PMID- 15877927 TI - Helping individuals to help themselves. AB - Obesity is a serious and increasing health issue. Approximately two-thirds of adults in the UK are now overweight or obese. Recent public health reports firmly reinforce the importance of engaging individuals to look after their health, including their weight. They also spell out the need for individuals to be supported more actively, on many levels, to enable this 'engagement'. Meanwhile, national surveys indicate that approximately two-thirds of adults are concerned about weight control, with one-third actively trying to lose weight. This finding is hardly surprising considering current weight statistics, plus the plethora of popular diets on offer. Weight-loss methods include diet clubs, diet books, exercise, meal replacements, advice from healthcare professionals and following a self-styled diet. Obesity is a multi-factorial problem, and losing weight and, in particular, maintaining weight loss is difficult and often elusive. It is argued that the modern obesogenic or 'toxic' environment has essentially taken body weight control from an instinctive 'survival' process to one that needs sustained cognitive and skill-based control. The evidence suggests that health professionals can help individuals achieve longer-term weight control by supporting them in making sustainable lifestyle changes using a range of behavioural techniques. These techniques include: assessing readiness to change; self-monitoring; realistic goal setting; dietary change; increased physical activity; stimulus control; cognitive restructuring; relapse management; establishing ongoing support. Consistently working in a client-centred way is also being increasingly advocated and incorporated into practice to help motivate and encourage, rather than hinder, the individual's progress. PMID- 15877928 TI - Weight management in primary care: how can it be made more effective? AB - Obesity is often difficult to tackle in primary care. Pressure of time in the consultation, a lack of appropriately-trained primary care staff, a shortage of community dietitians or nutritionists, the potentially enormous caseload, language or cultural barriers and the sheer intractability of patients' eating habits, exercise behaviour and their clinical condition, all conspire to make general practitioners, other team members and often the patients themselves lose heart and stop even trying. However, there are ways of overcoming these difficulties. Examples of changes that evidence suggests are able to support and enhance basic one-to-one interventions in general practice include: improved clinical guidelines; better training of primary care staff; at-risk patient registers; smarter database search tools; new quality incentives; closer working with dietitians, counsellors and pharmacists; more hospital outreach clinics; designated general practitioner specialists and practice clustering; expanded exercise referral schemes and links with leisure providers; subsidised referral to commercial slimming groups; better use of patient groups and voluntary and community workers. The present paper describes a proposed 'triple-tier' pathway for weight management incorporating most of the elements mentioned earlier. With a more joined-up and creative approach to the development and organisation of primary care, more comprehensive training and workforce planning, and better integration with social care, voluntary groups and the commercial sector, weight management in general practice has the potential to be much more effective. PMID- 15877929 TI - Low-glycaemic diets and health: implications for obesity. AB - The present review considers the background to terminology that relates foods, glycaemia and health, including 'available carbohydrate', 'glycaemic index' (GI), 'glycaemic glucose equivalent', 'glycaemic response index' and 'net carbohydrate', and concludes that central to each of these terms is 'glycaemic load' (GL). GL represents the acute increase in exposure of tissue to glucose determined by foods; it is expressed in ingested glucose equivalents (per 100 g fresh weight or per serving), and is regarded as independent of the state of glucose metabolism from normal to type 2 diabetes mellitus (T2DM). Ad libitum studies in overweight or obese adults and children show that low-GL diets are associated with marked weight benefits, loss of adiposity and reduced food intake. Weight benefits appear on low-glycaemic v. high-glycaemic available carbohydrates, unavailable v. available carbohydrates and protein v. available carbohydrate. Energy intake immediately after lowering of meal GL via carbohydrate exchanges is apparent only after a threshold cumulative intake of >2000 MJ. Various epidemiological and interventional studies are discussed. A relationship between GL and the development of T2DM and CHD is evident. Studies that at first seem conflicting are actually consistent when data are overlaid, such that diets with a GL of >120 glucose equivalents/d would appear to be inadvisable. Whereas certain studies might place GI as being slightly stronger than GL in relation to T2DM risk, this situation appears to be associated with observations in a lower range of GL or when the range of GI is too narrow for accuracy; nevertheless, authors emphasise the importance of GL. Among the studies reviewed, GL offers a better or stronger explanation than GI in various observations including body weight, T2DM in nurses, CHD, plasma triacylglycerols, HDL-cholesterol, high-sensitivity C-reactive protein and protein glycation. Where information is available, the associations between risk factors and GL are either similar or stronger in the overweight or obese, as judged by BMI, and apply to both body weight and blood risk factors. The implications tend to favour a long term benefit of reducing GL, for which further study is necessary to eliminate any possibility of publication bias and to establish results in clinical trials with overweight and obese patients. PMID- 15877930 TI - Expressing the glycaemic potency of foods. AB - The glycaemic index (GI) was introduced to guide food exchanges within equicarbohydrate food categories, and it expresses the glycaemic potency of the available carbohydrate component in a food relative to that of glucose. As GI is a relative value based on 'available carbohydrate' it cannot guide food choice for glycaemic control unless the foods are equal in available carbohydrate. Furthermore, GI cannot respond to food intake or to effects on food glycaemic potency of replacing glycaemic ingredients with non-glycaemic ingredients. The glycaemic glucose equivalent (GGE) overcomes these limitations of GI. The GGE content of an amount of food is the weight of glucose (g) that would induce a glycaemic response equal to that induced by the food. Few studies have compared GI and GGE as guides to food choice for glycaemic control, but in a direct test of the predictive validity of GGE in a group of foods of differing carbohydrate and GI, GGE predicted glycaemic potency well, whereas GI was unrelated to glycaemic effect. Furthermore, an information-processing model of the use of food information in food choice shows that GI has fundamental flaws when used outside the restriction of equicarbohydrate food exchange categories. As a general guide to food choices for the control of glycaemia GI does not satisfy the criteria predictive validity, accuracy, safety, ease of use, flexibility, sufficiency and compatability, whereas GGE does. GGE is also a scientifically precise and meaningful term with which to express glycaemic potency than is 'glycaemic load'. PMID- 15877931 TI - The role of plant secondary metabolites in mammalian herbivory: ecological perspectives. AB - Plant secondary metabolites (PSM) have many ecological functions, but have long been considered as defences against pathogens or herbivores (vertebrate or invertebrate), reducing the likelihood and extent of attack. However, mammalian herbivores ingest many foods containing PSM and use both behavioural methods and physiological strategies to limit their negative effects. Most physiological counter-adaptations are inducible in response to ingested PSM, providing efficient protection against toxic effects. Possible positive effects of PSM include antioxidant and anthelminthic properties and complex formation between protein and condensed tannins that protects dietary protein from degradation by the symbiotic microflora of foregut fermenters, increasing its utilisation by the animal. This protein effect is probably only beneficial to animals under a narrow range of nutrient-rich conditions found mainly in agricultural systems. There are many examples of PSM causing food avoidance or reducing food intake, but there is as yet relatively little evidence for positive selection of them by herbivores. Although the feedback mechanisms relating the post-ingestive consequences of PSM to subsequent foraging behaviour are beginning to be understood, knowledge of the integration of behavioural and physiological strategies for regulating the effects of PSM is relatively poor. The opportunities for learned avoidance of PSM may be restricted in animals with complex diets that cannot associate a particular feedback signal with a given food type. A greater emphasis on the study of subclinical effects of PSM rather than acute effects, on pharmaco kinetic studies in relation to behavioural studies and on the use of realistic experimental models is advocated. PMID- 15877932 TI - Muscarinic and nicotinic receptors synergistically modulate working memory and attention in humans. AB - Functional abnormalities in muscarinic and nicotinic receptors are associated with a number of disorders including Alzheimer's disease and schizophrenia. While the contribution of muscarinic receptors in modulating cognition is well established in humans, the effects of nicotinic receptors and the interactions and possible synergistic effects between muscarinic and nicotinic receptors have not been well characterized in humans. The current study examined the effects of selective and simultaneous muscarinic and nicotinic receptor antagonism on a range of cognitive processes. The study was a double-blind, placebo-controlled, repeated measures design in which 12 healthy, young volunteers completed cognitive testing under four acute treatment conditions: placebo (P); mecamylamine (15 mg) (M); scopolamine (0.4 mg i.m.) (S); mecamylamine (15 mg)/scopolamine (0.4 mg i.m.) (MS). Muscarinic receptor antagonism with scopolamine resulted in deficits in working memory, declarative memory, sustained visual attention and psychomotor speed. Nicotinic antagonism with mecamylamine had no effect on any of the cognitive processes examined. Simultaneous antagonism of both muscarinic and nicotinic receptors with mecamylamine and scopolamine impaired all cognitive processes impaired by scopolamine and produced greater deficits than either muscarinic or nicotinic blockade alone, particularly on working memory, visual attention and psychomotor speed. These findings suggest that muscarinic and nicotinic receptors may interact functionally to have synergistic effects particularly on working memory and attention and suggests that therapeutic strategies targeting both receptor systems may be useful in improving selective cognitive processes in a number of disorders. PMID- 15877933 TI - Increased phosphorylation of Ser473-Akt, Ser9-GSK-3beta and Ser133-CREB in the rat frontal cortex after MK-801 intraperitoneal injection. AB - GSK-3beta is regarded as playing an important part in the pathogenesis of schizophrenia and the action of psychotomimetic agents. We observed phosphorylation of molecules associated with the GSK-3beta signalling pathway in the rat brain after MK-801 injection, which induces a schizophrenia-like state in humans. Ser9-GSK-3beta phosphorylation was increased after injection of 1 mg/kg MK-801 in the rat frontal cortex but not in the hippocampus or cerebellum. This increase peaked at 30 min and was maintained until 90 min after injection. The phosphorylation showed a dose-dependent increase up to 1 mg/kg MK-801, followed by a decrease at higher dosage. Furthermore, phosphorylation of Ser473-Akt and Ser133-CREB showed similar temporal, dose-dependent and regionally specific patterns with those of Ser9-GSK-3beta. However, phosphorylation of Dvl and Ser33 beta-catenin was not affected by MK-801. These results suggest that GSK-3beta phosphorylation by MK-801 may be associated with the Akt-GSK-3beta pathway rather than with the Wnt-Dvl-GSK3beta pathway. PMID- 15877934 TI - Caffeine improves spatial learning deficits in an animal model of attention deficit hyperactivity disorder (ADHD) -- the spontaneously hypertensive rat (SHR). AB - The spontaneously hypertensive rat (SHR) is generally considered to be a suitable genetic model for the study of attention deficit hyperactivity disorder (ADHD), since it displays hyperactivity, impulsivity, poorly sustained attention, and deficits in learning and memory processes. Converging evidence suggests a primary role of disturbance in the dopaminergic neurotransmission in ADHD patients and in SHR, and in addition, some studies have also demonstrated alterations in adenosinergic neurotransmission in SHR. In the present study, adult female Wistar (WIS) and SHR rats received caffeine (1-10 mg/kg i.p.) 30 min before training, immediately after training, or 30 min before a test session in the spatial version of the Morris water maze. The effect of caffeine administration on WIS and SHR blood pressure was also measured. SHR needed significantly more trials in the training session to acquire the spatial information, but they displayed a similar profile to that of WIS rats in the test session (48 h later), demonstrating a selective deficit in spatial learning. Pre-training administration of caffeine (1-10 mg/kg i.p.) improved this spatial learning deficit in SHR, but did not alter the WIS performance. In contrast, post-training administration of caffeine (3 mg/kg i.p.) did not alter the SHR test performance, but increased memory retention in WIS rats. No dose of caffeine tested altered the mean blood pressure of WIS or SHR. These results demonstrate a selective spatial learning deficit in SHR which can be attenuated by pre-training administration of caffeine. In addition, the present findings indicate that the spatial learning deficit in SHR is not directly related to hypertension. PMID- 15877935 TI - The relationship between serum folate, vitamin B12, and homocysteine levels in major depressive disorder and the timing of improvement with fluoxetine. AB - The objective of the present study was to examine the relationship between serum folate, vitamin B12, and homocysteine levels and the timing of clinical improvement to fluoxetine in major depressive disorder (MDD) patients. A total of 110 outpatients with MDD who responded to an 8-wk trial of fluoxetine had serum folate, B12, and homocysteine measurements at baseline (prior to fluoxetine initiation). Onset of clinical improvement was defined as a 30% decrease in Hamilton Depression Scale scores that led to a 50% decrease by week 8. Patients with low folate levels (0.05). In conclusion, low serum folate levels were found to be associated with a delayed onset of clinical improvement during treatment with fluoxetine in MDD by, on average, 1.5 wk. PMID- 15877937 TI - A population-based study of the waiting times for prostatectomy in Ontario. AB - INTRODUCTION AND OBJECTIVE: Despite the high incidence of prostate cancer in Canada, there is currently limited information describing how these patients are being managed. The aim of this study was to review the surgical waiting times for radical prostatectomy in Ontario, utilizing existing population-based cancer databases, and to describe factors associated with prolonged waiting times. METHODS: This is a retrospective, population-based, observational study of men diagnosed with prostate cancer in Ontario between 1980 and 2000. The sources of data include the Ontario Cancer Registry linked to hospital discharge data, as well as census data from Statistics Canada. Study variables include age, county of residence, teaching hospital status, hospital surgical volume, area-level median household income and cause-specific survival. Waiting times were compared across study variables using univariate and graphical methods. Survival was compared across geographic regions with differing average wait times. RESULTS: We identified 9524 men treated with radical prostatectomy in Ontario over the study period and found the percentage of all patients with the disease who were treated surgically increasing from 3% to 20% over the last 2 decades. The overall time to prostatectomy has almost doubled with a median waiting time of 55 days in earlier eras to 91 days in 1996-2000. A few counties had significantly different wait times, whereas age and socio-economic factors were not associated with wait times across most eras. In the most recent eras, acute care hospitals and hospitals with higher surgical volumes had significantly higher waiting times (up to 20 days longer in 1996-2000, p<0.0001). Patients living in regions with the shortest wait times had statistically significant worse survival (p=0.02), implying that triaging has a greater impact than the potential effect of prolonged waits. CONCLUSIONS: The observed increases in waiting times for radical prostatectomy from this study are similar to the known increases in waiting times for radiotherapy. This increased time to treatment is an illustration of the stress on the health care system in Ontario. PMID- 15877938 TI - Compliance and outcome of patients with stage 1 non-seminomatous germ cell tumors (NSGCT) managed with surveillance programs in seven Canadian centres. AB - OBJECTIVE: We evaluate the impact of surveillance programs on the outcome of men with clinical stage 1 NSGCT following orchidectomy. PATIENTS AND METHODS: A retrospective review of 197 patients with a minimum of 2 years follow-up at seven cancer centres was conducted. Histological characteristics of the primary tumor were recorded for each patient. Surveillance protocols consisted of clinical assessments, chest X-rays, serum beta HCG (bHCG), alpha feto-protein (aFP), and abdominopelvic CT. All clinic visits and test completions were tracked. In accordance with each centre's specific surveillance protocol, patient compliance was defined as missing no more than two assessments/year. RESULTS: Overall 5 year survival was 100%. With a median follow-up of 54 months (range: 11-164 months), the relapse rate at 5 years was 29%. The median time to relapse was 6 months (range: 2-135 months). Ninety percent of relapses occurred within 18 months and only two patients relapsed after 5 years. On univariate analysis, only the presence of lymphovascular invasion was predictive of relapse. The first indicator of relapse was: CT alone, 36%; elevated bHCG or aFP, 29%; CXR, 10%; or clinical exam, 7%. Either CT, tumor markers, or CXR detected 90% of all relapses. Although differences in the frequency of assessments between the centres existed, no significant differences occurred in rates of relapse or survival (p>0.07). The mean rate of compliance with clinic visit (which included CXR and tumor markers) was 78% (range: 68.4-94.2%). The mean rate of compliance with CT scanning, was 64.3% (range: 32.2-100%). In the centre with the protocol requiring the least frequent visits, the rates of compliance were observed to be highest. CONCLUSIONS: Surveillance remains an effective means of managing stage 1 NSGCT despite variability in protocols and in patients compliance. An abnormal CT was the most frequent identifier of disease relapse, and in combination with tumor markers and CXR, 90% of relapses were detected within 2 years of orchiectomy. Modifications of surveillance protocols to less frequent assessments may be possible and should be subject to prospective evaluation. PMID- 15877939 TI - Pubovaginal bone anchor fixation with polyethylene versus fascia lata slings in the treatment of female stress incontinence: sling material and processing are predominant factors in success. AB - OBJECTIVE: The opponents of the In-Taca bone anchor system note the risk of a high rate of wound infection and osteitis pubis. We evaluated whether there is a difference in the outcome of the use of two different sling materials- polyethylene and fascia lata--with regard to wound infection, and analyzed the incidence of osteitis pubis further in a larger series. MATERIAL AND METHODS: A total of 61 women (mean age = 65.4 years) were treated for stress urinary incontinence (SUI) type II and III using the In-Taca bone anchor system. In 15 of 61 patients, we used a synthetic sling of polyethylene, and in 46, a fascia lata sling. The subjective success rate was determined with validated questionnaires (Urinary Distress Inventory-6, Symptom Severity Index and Symptom Impact Index). The objective assessment included a pad test according to the ICS- standard and a urogynecologic evaluation. Mean follow-up was 10.2 months. RESULTS: Wound inflammation of only very mild degree occurred in 15% in the fascia lata group, whereas 33% in the polyethylene group developed serious sling infection; in three patients explantation of the sling was necessary. Accordingly, satisfaction with the procedure was low in the polyethylene group. In both groups, there were no hints of osteitis pubis. The sling material used did not affect continence rate. CONCLUSION: Using the bone anchor system, the infection rate depends primarily on the sling material used and its processing: polyethylene is well tolerated in other reconstructive procedures (such as TVT, where a netlike mesh is used), so the processing of synthetic sling material plays an extremely important role in infection rate: platelike, dense synthetic material tends to cause wound infection. PMID- 15877940 TI - Validation of 1997 Partin Tables' lymph node invasion predictions in men treated with radical prostatectomy in Montreal Quebec. AB - OBJECTIVE: The accuracy of 1997 Partin Tables' lymph node invasion (LNI) predictions exhibits important variability in different testing populations. We explored the LNI predictive accuracy in radical prostatectomy (RP) patients from Montreal, Canada. Moreover, we assessed the extent of change in predictive accuracy related to a modification of PSA coding from categorical to continuous. METHODS: We used pretreatment serum PSA, clinical stage, and biopsy Gleason sum from 537 men treated with RP to compare predicted and observed rates of LNI. Accuracy was quantified with receiver-operating characteristics curves. RESULTS: Accuracy was 0.760 in 369 evaluable patients, when categorically coded pretreatment PSA (0-4, 4.1-10, 10.1-20, 20.1+) was combined with clinical stage and biopsy Gleason sum. A 2.7% accuracy increase was noted when categorically coded PSA was replaced with continuously coded values. CONCLUSION: Partin Tables' LNI predictions showed comparable accuracy to a community-based sample from the United States (0.766), and to a recent, multi-institutional sample (0.740). However, accuracy was lower than reported in internal (0.818), and external (0.837) academic, validation cohorts. Accuracy of LNI predictions was appreciably higher, when continuously coded PSA was used. PMID- 15877941 TI - Technology review: high-intensity focused ultrasound for prostate cancer. AB - INTRODUCTION AND OBJECTIVE: High-Intensity Focused Ultrasound (HIFU) is a technology that has moved from being used for benign prostate disease to the treatment of prostate cancer. A technology review was undertaken to guide patients and physicians as to its suitability. METHOD: An evidence-based review of published papers in the English language, with additional material from internet and other sources. RESULTS AND CONCLUSIONS: Only case series have been published; there are no randomized studies. The quality of evidence is poor, with no reports having longer follow-up than a mean of 2 years, with median follow-ups substantially shorter. Efficacy outcomes are thus premature and preclude assessment. Toxicity varies substantially with impotence rates 44%-61%, grade 2-3 incontinence 0%-14%, and rectal fistulae 0.7%-3.2%. There is limited data on the use of HIFU for the salvage therapy after radiation failure. There are no data on the toxicity of subsequent standard curative therapies after HIFU. In view of the lack of efficacy outcomes, and in the presence of significant toxicity, HIFU should only be offered within a research setting. PMID- 15877942 TI - Phimosis--a diagnostic dilemma? AB - INTRODUCTION: Phimosis is defined as the inability to retract the foreskin. Differentiating between physiological phimosis and pathological phimosis is important, as the former is managed conservatively and the latter requires surgical intervention. Referrals of patients with physiological phimosis to urology clinics may create anxiety regarding the need for surgery amongst patients and parents, while unnecessarily expanding the waiting list for specialty assessment. OBJECTIVES: To determine the ability of referring physicians to differentiate physiological from pathological phimosis, and to see whether there is any difference in this ability between generalists versus specialists. MATERIALS AND METHODS: A retrospective chart review of 284 consecutive referrals for phimosis to the Children's Hospital of Eastern Ontario (CHEO) Urology Clinic during November 2000 - April 2003 was conducted. Referral sources included family physicians (FP), pediatricians (PD), emergency physicians (ER), and other subspecialists (SS). Data for this study were obtained from the original referral letters and cross-referenced with the impressions of the pediatric urologist following the initial patient encounter. The accuracy in diagnosing phimosis was evaluated among the various types of referring physicians. RESULTS: A total of 284 phimosis referrals were reviewed of patients ranging from 2 months to 16 years of age (mean = 6.6 years). The referral sources consisted of 222-GP, 33-PD, 23-ER, and 6-SS. The majority of referred cases were diagnosed by the attending pediatric urologist as physiological phimosis across all referral sources, with the exception of subspecialists (FP = 75.2%, PD = 81.8%, ER = 56.5%, SS = 33.3%). Second to this was the diagnosis of pathological phimosis across all referral sources except SS (FP = 14.9%, PD = 12%, ER = 34.8%, SS = 50%). Overall, the circumcision rate for the 284 phimosis referrals reviewed was 14.4%. CONCLUSIONS: Our findings reveal that many physicians continue to face difficulties in distinguishing physiological phimosis from the pathological. As a result, many unnecessary referrals are made for phimosis . We suggest the implementation of improved educational measures regarding preputial pathophysiology in the medical curriculum. Such measures would serve two purposes: first, to reduce the number of unnecessary specialty referrals and secondly, to aid primary care physicians in recognizing the presence of physiological phimosis so that patients and families may be reassured of normalcy. PMID- 15877943 TI - Primary small cell carcinoma of the ureter. AB - Primary small cell carcinoma of the ureter is a rare clinico-pathological entity. We present a report of primary ureteric small cell carcinoma and pathological correlates. PMID- 15877944 TI - Understanding participation in a trial comparing cryotherapy and radiation treatment. AB - BACKGROUND: To date, few two-arm active treatment randomized control trials (RCTs) have compared prostate cancer therapies. OBJECTIVE: To examine the difference and similarities between the reasons for accepting and declining participation in a two-arm active treatment RCT comparing external beam radiation therapy (EBRT) versus cryotherapy. METHODS: Eleven men with prostate cancer, selected purposively, participated in a 30-minute post-treatment semi-structured interview. Interviews were transcribed verbatim, coded and analyzed for patterns with the assistance of the text management (TM) software (NVivo). RESULTS: RCT accepters participated principally with the hope of being randomized into the cryotherapy treatment arm. Consequently, randomization into the EBRT arm was often perceived as receiving the consolation prize. RCT decliners were either pushed away from cryotherapy and/or pulled towards another treatment (surgery, EBRT, brachytherapy). Factors influencing accepters'/decliners' treatment decisions include (1) personal factors such as patient research and treatment preference, cancer survivors, family/friends, and altruism, and (2) physician, trial, and treatment factors such as patient-physician rapport, RCT awareness and understanding, therapy convenience, expected outcome and perceived side effects. CONCLUSIONS: By better understanding patients' views about RCT participation, recruitment rates for prostate cancer RCTs can be improved. PMID- 15877945 TI - Mucocolpos associated with bladder exstrophy: a case report. AB - INTRODUCTION: We present the first reported case demonstrating an association of mucocolpos and bladder exstrophy. MATERIALS: A term baby girl presented with bladder exstrophy and underwent a successful primary bladder closure and ureteral reimplantation. Subsequently, she presented with a fluid-filled pelvic mass which was found to be a mucocolpos. RESULTS: Surgical drainage of the collection was required. CONCLUSION: Mucocolpos should be considered in the differential diagnosis of a pelvic mass in a patient with bladder exstrophy. PMID- 15877946 TI - [Scanning 2005 international consensus on guideline for cardiopulmonary resuscitation and emergency cardiovascular care (2)--basic life support play a key and important role in cardiopulmonary resuscitation]. PMID- 15877947 TI - [Clinical trial and evaluation on comprehensive treatment on attack in acute stage: report of 522 cases]. AB - OBJECTIVE: To investigate the clinical effect of the comprehensive treatment to acute stage of attack. METHODS: On the basis of the previous observation, the study of the randomization control with general treatment, treatment on acupuncture and western medical treatment were carried out. A comprehensive treatment on overall traditional Chinese medical differentiation according to the superiority of every treatment was assessed and evaluated in 522 patients with attack. RESULTS: The comprehensive treatment of cerebral infarction was superior to the western medicine treatment. General treatment, treatment on traditional Chinese medical differentiation, acupuncture group revealed different improvement on neural function, daily viability, cognitive function in various extent. CONCLUSION: The comprehensive treatment that based on overall traditional Chinese medical differentiation has advantage and characteristic. It has positive combined action to the attack and relevant to clinical setting, easier to popularization and application. Various appraising amount form has different evaluating effects in different stage. PMID- 15877948 TI - [Expression of heat shock protein 70 in intestinal mucosa during the early stage after severe burns and its significance]. AB - OBJECTIVE: To study the expression of heat shock protein 70 (HSP70) in intestinal mucosa during the early stage after severe burn injury and its significance. METHODS: With a model of 30% total body surface area (TBSA) full-thickness burned rats, the expression and distribution of HSP70 and heat shock factor-1 (HSF1) in intestinal mucosal were determined by reverse transcription-polymerase chain reaction (RT-PCR), Western blot and immunohistochemistry at 3, 6, 12, 24 and 48 hours postburn. RESULTS: The expression of HSP70 mRNA and protein in intestinal mucosa increased markedly at 3 hours after severe burns, peaked at 6 and 12 hours, and lasted for 48 hours postburn (all P<0.01). Following a slight decline at 3 hours postburn, the expression of HSF1 increased gradually, and reached a highest level at 48 hours postburn (all P<0.01). CONCLUSION: The expression of HSP70 and HSF1 markedly increase in intestinal mucosa following severe burn injury. It is suggested that the self-protective mechanism of cells might explain the increase of HSP70 expression. PMID- 15877949 TI - [Change in apoptosis and its mechanism in intestinal epithelial cells under oxidative stress]. AB - OBJECTIVE: To investigate change in apoptosis level and its mechanism of intestinal epithelial cells under oxidative stress. METHODS: HT-29 cells were cultured in vitro, which were treated with hydrogen peroxide (H2O2), to simulate the intestinal epithelial cells injured by reactive oxidative species. The cells viability was observed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Moreover, cell apoptosis and apoptosis associated proteins were evaluated by flow cytometry and Western blot. RESULTS: Cells viability of HT 29 was decreased by H2O2 which showed dose-dependent and time-dependent patterns (all P<0.05). The cell apoptotic ratios were increased with the concentration of H2O2 increased and the time of the stimulation prolonged compared with the controls (all P<0.05). Although the expression of the Bax was increased when HT 29 cells were stimulated with different concentrations of H2O2 for 24 hours, the expression of Bcl-2 was decreased. While HT-29 cells were stimulated with 500 micromol/L H2O2, the expression of the Bax was increased and that of Bcl-2 was decreased overtime. CONCLUSION: These data suggest that oxidative stress appears to be related to the apoptosis in intestinal epithelial cells under stress. The imbalance of Bcl-2/Bax expression might result in intestinal epithelial cell apoptosis in oxidative stress. PMID- 15877951 TI - [Study on high mobility group-1 protein in patients with multiple trauma]. AB - OBJECTIVE: To study the change in serum high mobility group-1 (HMG-1) protein in patients with multiple trauma and determine its relationship to severity of trauma and multiple organ dysfunction syndrome (MODS). METHODS: In 33 cases, HMG 1 levels were determined on 1, 3, 7 days after trauma respectively. The values of injury severity score (ISS), acute physiology and chronic health evaluation II (APACHE II) and systemic inflammatory response syndrome (SIRS) score were determined. Meanwhile, the occurrence of organ dysfunction was noted. RESULTS: The level of serum HMG-1 in patients with multiple trauma was much higher than that of normal controls, and also in those with organ dysfunction. HMG-1 and APACHE II were positively correlated (r=0.495, P=0.016). The level of serum HMG-1 and APACHE II value were much higher in patients with organ dysfunction than those of normal organ function, but there was no significant difference in SIRS score between the two groups (P=0.105). CONCLUSION: Serum HMG-1 increases in patients with trauma and is positively correlated with severity of trauma. Serial determination of HMG-1 is helpful to discover clinical infection earlier. HMG-1 can be used as a warning indicator of the onset of MODS. PMID- 15877952 TI - [Effects of injecting heparin into the center of hematoma on peri-hematoma edema and hematoma volume in pigs with intracerebral hemorrhage]. AB - OBJECTIVE: To investigate the safety and effect of injecting heparin into hematoma on peri-hematoma edema and hematoma volume in pigs with intracerebral hemorrhage (ICH). METHODS: Thirteen sucking pigs were divided randomly into two groups: hemorrhage group, in which 2.5 ml arterial blood was injected into the right frontal lobe and heparin group, in which 0.2 ml of heparin was injected into the hematoma produced by the injection of 2.3 ml of blood into the similar site. The hematoma volume and peri-hematoma edema were determined by the sequences of T2* weighted image (T2*WI), fluid-attenuated inversion- recovery (FLAIR) image and diffusion weighted image (DWI) by 1.5 T magnetic resonance image (MRI) from 30-60 minutes afterwards to 24 hours. The peri-hematoma apparent diffusion coefficient (ADC) was compared with that of contralateral hemisphere, and the corresponding histologic changes were studied. RESULTS: The average volume, shown by T2*WI at 24 hours, was significantly larger than that at 30-60 minutes after hematoma formation in hemorrhagic group [(5.29+/-0.98) cm3 vs. (3.09+/-0.38) cm3, P<0.01]. But there was no significant change in hematoma volume in hemorrhagic group from 30-60 minutes on to 24 hours [(2.21+/-0.28) cm3 vs. (2.33+/-0.30) cm3, P>0.05]. Both increased and decreased ADC were found around the hematoma in some animals of the heparin group compared with that of the contralateral hemisphere. On the other hand, in hemorrhagic group, only increased ADC could be found around the lesion, and there was no decreased ADC. CONCLUSION: Injection of heparin into an intracerebral hematoma leads to enlargement of the hematoma and more marked peri-lesion edema. On ADC maps, enlargement of hematoma is attributed to the edema around the lesion leading to injury to the brain tissue. PMID- 15877953 TI - [Investigation on the relation between systemic inflammatory response syndrome and severity of acute pancreatitis]. AB - OBJECTIVE: To explore the relationship between the systemic inflammatory response syndrome (SIRS) and the severity of the acute pancreatitis, so as to provide some leads in the treatment to lower the mortality of severe acute pancreatitis. METHODS: One hundred and seventy-two patients suffering from acute pancreatitis admitted to our departments from January 1, 2001 to December 31, 2003 were retrospectively analyzed. The patients were divided into survivor group and non survivor group, and patients in the survivor group were further divided into severe group and mild group according to scores of acute physiology and chronic health evaluation II (APACHE II). The scores of APACHE II were compared between patients without SIRS and those who met the criteria of SIRS. RESULTS: There was significant difference between the survivor group and non-survivor group not only in the incidence of patients who developed SIRS, but also in the number of patients fulfilling or not fulfilling the diagnostic criteria of SIRS between the severe group and mild group. There was also significant statistical difference between the SIRS group and non-SIRS group in the APACHE II scoring. Significant difference in duration of SIRS was found between the severe group and mild group. CONCLUSION: SIRS is highly correlated with the severity of acute pancreatitis. Active prevention and treatment of SIRS may raise the survival rate of severe acute pancreatitis. PMID- 15877954 TI - [Evaluation of electroencephalography grading standard in severe cerebrovascular disease]. AB - OBJECTIVE: To compare the prognostic value of different electroencephalography (EEG) grading standards of cerebral dysfunction after cerebrovascular diseases. METHODS: The EEG of all the 40 patients who suffered from acute cerebrovascular disease of the cerebral hemisphere were recorded continuously, and three different EEG grading standards (Lavizzari, Synek and Young) were analyzed and compared. RESULTS: All three different EEG grading standards showed significant correlation with outcome (all P<0.001), i. e. the higher the EEG grade was, the poorer the outcome was. Logistic regression showed that the Synek standard was the best for survival rate and comprehensive prognostic accuracy (90.9%, 89.2%, respectively), Young standard (81.8%, 83.8%) was lower than Synek standard in predicting accuracy but higher than Lavizzari standard (75.%, 80.% respectively). CONCLUSION: The EEG grading standard of Synek might well reflect the cerebral dysfunction and appear to be a reliable predictor for outcome. PMID- 15877956 TI - [Experimental study of apoptosis and Fas antigen expression in craniocerebral trauma]. AB - OBJECTIVE: To evaluate the change in Fas antigen expression and apoptosis of neurons, to provide an experimental evidence of loss of neurons in craniocerebral injury, and to provide experimental evidence for clarifying multi-approach of the lost neuron after damage. METHODS: Brain impact injury was reproduced in SD rat with a free falling impacting device. Using immunohistochemistry method the Fas protein expression was assessed and apoptotic cells were detected with electron microscopy. RESULTS: Apoptotic cells were found near the contused area, and Fas positive cells appeared around the injured and hippocampus areas at about 4 hours, peaked at 24 hours after injury and then reduced in number. CONCLUSION: Apoptosis and necrosis are two forms of cell death in brain tissue following experimental brain contusion. Moreover, the results imply that the Fas-FasL pathway plays a pivotal role in the pathophysiology of post-traumatic neuronal apoptosis. PMID- 15877958 TI - [Effect of clausenamide on the expression of Bcl-2 protein and apoptosis after focal cerebral ischemia/reperfusion in renovascular hypertensive rats]. AB - OBJECTIVE: To observe the effect and mechanism of clausenamide on the expression of Bcl-2 and apoptosis after focal cerebral ischemia/reperfusion in renovascular hypertensive rats. METHODS: Seventy-five renovascular hypertensive rats were randomly divided into three groups (25 in each group): clausenamide intervention group, single ischemia/reperfusion model group and sham-operated group. Focal cerebral ischemia was reproduced by ligature for 2 hours and loosening of the ligature in the rats. No arterial ligature was applied in sham-operated group. Computerized pathological image analyzer was used to determine the number of cells positive for Bcl-2 by immunohistochemical staining, and also the counts of apoptotic cells after TdT-mediated dUTP nick end labeling (TUNEL) staining respectively in coronal sections of brain after reperfusion (6, 12, 24, 48 and 72 hours). RESULTS: (1) The expression of Bcl-2 protein was detected 6 hours after reperfusion, peaking at 24 hours, then declined gradually. The Bcl-2 protein positive cell counts at every time point in clausenamide intervention group were significantly higher than simple ischemia/reperfusion model group (all P<0.01). (2) The number of apoptotic cells was increased with reperfusion, reaching its peak at 72 hours. The apoptosis counts in clausenamide intervention group were significantly lower than single ischemia/reperfusion model group (all P<0.01). At all time points, except at 48 hours after reperfusion, as there was no significant difference (all P>0.05). No Bcl-2 positive cells and only 0-2 apoptotic cells could be discernible in brain sections from sham-operated animals or in the contralateral side of ischemia in animals of the other groups. CONCLUSION: Expression of Bcl-2 protein is enhanced and apoptosis appears after focal cerebral ischemia/reperfusion in rat brain. Clausenamide can enhance the expression of Bcl-2 protein and inhibit apoptosis remarkably. Clausenamide may coordinate with Bcl-2 in inhibiting apoptosis. This may be the mechanism of protection of brain cells from ischemic damage of clausenamide treatment. PMID- 15877960 TI - [Correlation of plasma endothelin and multiple organ dysfunction syndrome caused by acute paraquat poisoning]. AB - OBJECTIVE: To investigate the relationship between endothelin (ET) and multiple organ dysfunction syndrome (MODS) caused by acute paraquat poisoning (APP). METHODS: The levels of plasma ET were measured by radioimmunoassay in 24 patients with MODS caused by APP and 19 healthy persons as controls. The levels of plasma ET patients in MODS caused by APP were correlatively analysed with acute physiology and chronic health evaluation II (APACHE II) score, partial pressure of oxygen in artery (PaO2), troponin I (cTnI) level, blood enzymology and biochemistry indexes. RESULTS: The levels of plasma ET in patients with MODS caused by APP were much higher than controls (P<0.01), the level of plasma ET in death group was much higher than that of survivor group (P<0.01). Plasma ET was positively correlated with APACHE II score, cTnI, MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN) and serum creatinine (SCr, P<0.05 or P<0.01), where there was a negative correlation with PaO2 (P<0.01). CONCLUSION: Endothelin may be involved in the pathogenesis of MODS caused by APP, plasma ET may be one of a clinical index for evaluating the degree of multiple organ function damage, for guiding treatment, and judging the prognosis of MODS caused by APP. PMID- 15877961 TI - [Effect of aloe vera polysaccharide on the release of cytokines and nitric oxide in cultured human keratinocytes]. AB - OBJECTIVE: To investigate the effects of polysaccharide extracted from Aloe Barbadensis on the release of cytokines and nitric oxide (NO) in cultured human keratinocytes. METHODS: The levels of transforming growth factor-alpha (TGF alpha), TGF-beta1, interleukin-1beta (IL-1beta), IL-6, IL-8, tumor necrosis factor (TNF) and NO in the supernatants of keratinocyte culture in which culture media containing 25, 50, 100, 200, 400 microg/ml, respectively of aloe polysaccharide were assayed. In the control group equal volume of media without the polysaccharide was used. RESULTS: Compared with control group, the levels of TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF in the supernatants of cultured keratinocytes were significantly higher when aloe polysaccharide was added (P<0.05 or P<0.01), and they were positively correlated to the concentration of aloe polysaccharide (P<0.01). However, aloe polysaccharide markedly decreased the level of NO in a dose dependent manner (P<0.01). CONCLUSION: Aloe polysaccharide could promote keratinocytes to secrete TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF, and inhibit the release of NO. PMID- 15877962 TI - [Expression of heat shock protein 72 in leukocytes from patients with acute trauma and its relationship with survival]. AB - OBJECTIVE: To determine the expression of heat shock protein 72 (HSP 72) in leukocytes of patients with acute trauma, and evaluate its relationship with survival. METHODS: Thirty acute trauma patients with acute physiology and chronic health evaluation III (APACHE III) > or = 18 were included as study subjects. Blood samples were serially obtained at three time points: days 0-1, days 2-6 and days 7-14 after the trauma. Expression of HSP72 was assessed with immunohistochemistry, and the expression of HSP72 mRNA was assessed with semi quantified reverse transcription-polymerase chain reaction (RT-PCR), and the relationship between their levels and the survival rate (survival period > 2 months) was analyzed. RESULTS: The expression of HSP72 was significantly enhanced on post-trauma days 2-6. The levels of HSP72 in the survivor groups were higher than that in non-survivors. CONCLUSION: The study suggests that acute trauma can lead to a marked elevation of HSP72, and its peak level appears on days 2-6, which might be correlated with 2-month survival of the afflicted patients. PMID- 15877963 TI - [Role of trefoil peptides in modulation of gastric adaptation to stress]. AB - OBJECTIVE: To investigate the role of trefoil peptides in modulation of gastric adaptation to water restraint stress (WRS) in rats. METHODS: Wistar rats were exposed to single or repeated WRS for 4 hours every other day for up to 6 days, gastric mucosal blood flow (GMBF) was measured by LDF-3 flowmeter, the extent of gastric mucosal lesions was evaluated grossly and histologically, and expression of PS2 intestinal trefoil peptide (ITF), cyclooxygenase (COX-2), inducible nitric oxide synthase (iNOS) and transferase growth factor-alpha (TGF-alpha) were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: One application of WRS produced extensive gastric mucosal erosion. With repeated WRS, the gastric mucosa became adapted to the stress of WRS, and the ulcerative index (UI) was reduced by 22.0% that of one WRS challenge after four consecutive WRS. Expression of PS2 was markedly decreased and expression of ITF, COX-2, iNOS and TGF-alpha were markedly increased after single stress. But this adaptation to WRS was accompanied by increased GMBF and active cell proliferation in the neck region of gastric glands, and by increased expression of PS2, ITF, TGF-alpha, but reduced expression of COX-2 and iNOS. CONCLUSION: Gastric adaptation to WRS injury involves enhanced cell proliferation, increased expression of PS2, ITF, TGF-alpha and reduced expression of COX-2 and iNOS, suggesting trefoil peptides might play an important modulating role in this phenomenon. PMID- 15877977 TI - The effects of response priming on the planning and execution of goal-directed movements in the presence of a distracting stimulus. AB - Two models of selective reaching have been proposed to account for deviations in movement trajectories in cluttered environments. The response vector model predicts movement trajectories should deviate towards or away from the location a distractor of little or large salience, respectively. In contrast, the response activation model predicts that a distractor with large salience should cause movement deviations towards it whereas a distractor with little salience should not influence the movement. The precuing technique was combined with the distractor interference paradigm to test these predictions. Results indicate that when the target was presented at the precued (salient) location, movements were unaffected by a distractor. Conversely, when the distractor was presented at the precued location while the target was presented at an uncued (non-salient) location, participants demonstrated increased reaction times and trajectory deviations towards the location of the distractor. These findings are consistent with the model of response activation. PMID- 15877978 TI - The effect of visuo-motor transformations on hand-foot coordination: evidence in favor of the incongruency hypothesis. AB - Understanding how multiple constraints contribute to the emergence of coordinated behavior has been the topic of considerable debate in cognitive sciences. The present experiment addressed the issue of the effects of visual motion structures (iso- and non-isodirectionality) on the stability of hand-foot coordination patterns. Visuo-motor transformations--decorrelating the perceived movement direction from the actually generated direction--were applied to both in-phase and anti-phase patterns. Two mutually exclusive hypotheses--the "visual grouping hypothesis" and the "incongruency hypothesis"--were tested. The results indicated that both conditions of transformed visual feedback destabilized the actual performed coordination patterns. Thus, despite the existence of common underlying principles that govern both the perceived motion pattern and the generation of hand-foot coordination patterns, it appeared that perceptual grouping principles were not exploited to monitor the production of coordination. These results strongly suggest that the congruency between the performed pattern and the perceived visual feedback is the primary factor determining the (in)stability of hand-foot coordination patterns. PMID- 15877979 TI - Timed picture naming norms for 590 pictures in Dutch. AB - The present study presents timed norms for 590 pictures in Belgian Dutch. We determined name agreement and response latencies. Furthermore, we assessed which factors influenced the naming latencies of the pictures. It appeared that age-of acquisition, the H-statistic (an index of name agreement), and the number of syllables of the dominant response were significant predictors of the naming latencies. These results are discussed in comparison with previous findings. PMID- 15877980 TI - Response monitoring and expectancy in random serial RT tasks. AB - Two different sequential patterns have been reported in serial reaction-time (RT) tasks with random stimulus sequences, depending on the response-to-stimulus interval (RSI). When RSI is short a first-order repetition effect and a higher order benefit-only pattern are found, and when RSI is long a cost-benefit pattern is observed. In a series of three experiments, we found that the patterns are not unique for spatial dimensions, and that the benefit-only pattern does not appear for irrelevant stimulus features. The cost-benefit pattern, on the other hand, can shift to irrelevant locations, but not to irrelevant colour. Distributional analyses of the sequential effects demonstrate that the first-order repetition effect can be dissociated from the higher order effects. The results are interpreted as support for the role of response monitoring in the development of the benefit-only pattern at short RSI, while the first-order repetition effect is explained as repetition priming. With a long RSI, the cost-benefit pattern is confirmed as a low-level subjective expectancy process, which is primarily controlled by stimulus location. The data clearly strengthen the support for three different processes generating sequential effects in random serial RT tasks. PMID- 15877981 TI - The effects of action video game experience on the time course of inhibition of return and the efficiency of visual search. AB - The ability to efficiently search the visual environment is a critical function of the visual system, and recent research has shown that experience playing action video games can influence visual selective attention. The present research examined the similarities and differences between video game players (VGPs) and non-video game players (NVGPs) in terms of the ability to inhibit attention from returning to previously attended locations, and the efficiency of visual search in easy and more demanding search environments. Both groups were equally good at inhibiting the return of attention to previously cued locations, although VGPs displayed overall faster reaction times to detect targets. VGPs also showed overall faster response time for easy and difficult visual search tasks compared to NVGPs, largely attributed to faster stimulus-response mapping. The findings suggest that relative to NVGPs, VGPs rely on similar types of visual processing strategies but possess faster stimulus-response mappings in visual attention tasks. PMID- 15877982 TI - Importance of the postdischarge interval in assessing major adverse clinical event rates following percutaneous coronary intervention. AB - In-hospital major adverse clinical event (MACE) rates after percutaneous coronary intervention serve as benchmarks of performance. However, accelerated clinical pathways, decreased lengths of stay, and potential delayed effects of percutaneous coronary intervention may result in an underestimation of this traditional measurement of outcome. Records from patients in the first 3 waves of the National Heart, Lung, and Blood Institute's Dynamic Registry (n = 6,676) were reviewed for rates of composite in-hospital MACEs (death, myocardial infarction, and any repeat target vessel revascularization) and postdischarge MACEs (death, myocardial infarction, repeat hospitalization, and repeat target vessel revascularization) through 30 days. Rates for each composite MACE were compared across waves to assess changes over time. Predictors of each MACE category were identified using multivariate analysis. In-hospital MACE decreased significantly (5.4% of wave 1, 4.9% of wave 2, 3.1% of wave 3, p <0.001), whereas stent implantation increased significantly (67.5% of wave 1, 79.1% of wave 2, 86.2% of wave 3, p <0.001). Postdischarge MACE through 30 days remained unchanged (5.1% of wave 1, 5.1% of wave 2, 4.8% of wave 3, p = 0.6). Mean length of stay decreased (2.7 days for wave 1, 2.2 days for wave 3, p <0.001). Disparate clinical, procedural, and angiographic factors were associated with each MACE. Postdischarge MACE rates through 30 days comprise a significant and unchanging fraction of overall procedurally related MACE rates despite improving in-hospital outcomes. Most postdischarge events derive from pathology related to the controlled vessel. A 30-day MACE rate may serve as a more comprehensive measurement of procedural outcome. PMID- 15877983 TI - Comparison of thrombosis and restenosis risk from stent length of sirolimus eluting stents versus bare metal stents. AB - Selection of coronary stent length varies from covering only the zone of maximum obstruction to stenting from normal- to normal-appearing vessels. With bare metal stenting, for any given lesion there is a high restenotic risk associated with longer stent length. With drug-eluting stents, the relation between stent length and restenosis has not been evaluated. In the angiographic follow-up cohort of the SIRIUS trial that compared the sirolimus-eluting Bx Velocity stent with the standard Bx Velocity stent (n = 699), we constructed a multiple regression model to predict 8-month percent diameter stenosis using the main effects of lesion length and excess stent length beyond the lesion length and adjusting for known predictors of restenosis. Stent length exceeded lesion length in 94% of lesions overall. Mean difference in length was 8.3 +/- 8.3 mm (mean lesion length 14.6 +/ 5.9 mm, mean stent length 22.9 +/- 9.6 mm). Stented lesion length and excess stent length were associated with absolute increases in percent diameter stenosis per 10 mm of 9.1% (p <0.0001) and 3.6% (p = 0.053) in the bare metal arm and 3.5% (p = 0.047) and 2.1% (p = 0.040) in the sirolimus-eluting stent arm. Although the effects of lesion length and excess stent length on restenosis were markedly decreased with sirolimus-eluting stents (vs bare metal stents), a small restenotic penalty is still paid for excessive stent length. Longer stent-to lesion length strategies should be used only when a shorter stent is likely to result in incomplete lesion coverage and edge dissection, a strong determinant of stent thrombosis. PMID- 15877984 TI - Drug-eluting stents versus bare metal stents in percutaneous coronary interventions (a meta-analysis). AB - This meta-analysis combined the results of randomized clinical trials to compare the efficacy of drug-eluting stents with that of bare metal stents in percutaneous coronary interventions to ascertain which revascularization strategy is most safe and effective. The literature identified 13 published studies, and 8 were included in the main meta-analysis, thus allowing a meta-analysis on 3,860 patients for the effect on all major adverse clinical events (MACEs) combined and for target vessel revascularization. Meta-analyses were performed for combined MACEs, patient MACEs, and thrombosis. Regression meta-analyses were performed to examine the effect of certain variables on the efficacy of drug-eluting stents compared with bare metal stents. Meta-analysis of all trials showed that drug eluting stents produced significant decreases in the need for percutaneous revascularization (relative risk [RR] 0.30, 95% confidence interval [CI] 0.22 to 0.40) and coronary artery bypass grafting (RR 0.54, 95% CI 0.32 to 0.89). Drug eluting stents significantly decreased all MACEs combined (RR 0.40, 95% CI 0.33 to 0.49) but were not associated with an increased risk of stent thrombosis or death. These results were confirmed at analysis as stratified by type of eluting stent, because the need for percutaneous revascularization was significantly lower for sirolimus-eluting stents (RR 0.23, 95% CI 0.15 to 0.35) and paclitaxel eluting stents (RR 0.39, 95% CIl 0.29 to 0.53). PMID- 15877985 TI - Risk factors, echocardiographic patterns, and outcomes in patients with acute ventricular septal rupture during myocardial infarction. AB - Ventricular septal rupture (VSR), which can complicate an acute myocardial infarction (MI), carries a high mortality rate. Because precordial and transesophageal echocardiography can identify the type of rupture and assess right ventricular (RV) function at the patient's bedside, we examined the prognostic significance of echocardiographic patterns in postinfarct VSR by postulating that complex rupture and RV involvement carry a worse prognosis. Seventeen patients (10 men; mean age 66 years) who had confirmed postinfarct VSR underwent precordial and transesophageal echocardiography followed by coronary angiography. Serial 12-lead and right precordial leads were also available. Type of septal rupture was classified as simple or complex based on autopsy-proved echocardiographic criteria. Three patients had inferior wall MI and 14 had anterior wall MI. ST-segment elevation persisted >72 hours in all 3 patients who had inferior wall MI and in 12 who had anterior wall MI. Segmental wall motion abnormalities helped in detecting the left ventricular entry site, and use of unconventional views superimposed with color flow Doppler provided the RV exit site. RV function was better appreciated with transesophageal echocardiography. Two patients who had inferior wall MI and 7 who had anterior wall MI had complex ruptures. All 3 patients who had inferior wall MI and 7 who had anterior wall MI had electrocardiographic and echocardiographic evidence of RV involvement. Mortality rate was higher in patients who had complex rupture (78% vs 38%, p <0.001) and in those who had RV extension (71% vs 29%, p <0.001). In conclusion, persistent ST elevation is a common finding in patients who have postinfarct VSR. Complex VSR and RV involvement are significant determinants of clinical outcome. PMID- 15877986 TI - Heart rate response during dipyridamole stress as a predictor of mortality in patients with normal myocardial perfusion and normal electrocardiograms. AB - Although it is well established that a blunted chronotropic response to exercise is associated with a higher risk of death, recent data suggest a similar association between mortality risk and blunted heart rate response to vasodilatory stress. We investigated the heart rate response to dipyridamole induced stress as a predictor of death in the setting of normal myocardial perfusion and a normal electrocardiogram. We followed 1,087 patients for 8 years (range 5.7 to 11.8) who underwent dipyridamole vasodilator stress and had normal perfusion scans and electrocardiograms. None had heart failure, known left ventricular systolic dysfunction, pacemaker implantation, or valve disease. Heart rate response was assessed as the ratio of heart rate at peak stress to heart rate at rest. The primary end point was all-cause mortality. Quartile values for the peak-to-rest heart rate ratio were <1.19, 1.19 to 1.30, 1.31 to 1.44, and >1.44. There were 246 deaths. Death rates according to quartiles of heart rate ratio were 103 of 271 (38%), 64 of 272 (24%), 52 of 272 (19%), and 27 of 272 (10%). After adjusting for age, gender, heart rate at rest, blood pressure response, standard cardiovascular risk factors, and other confounders, a blunted heart rate response remained predictive of death (adjusted hazard ratio for lowest vs highest quartile 3.3, 95% confidence interval 2.1 to 5.1, p <0.0001). When considered as a continuous variable, the logarithm of the heart rate ratio was the strongest predictor of death, aside from age. Thus, among patients who have normal myocardial perfusion and normal electrocardiograms, a blunted heart rate response to vasodilator stress is predictive of a marked increase in risk of death. PMID- 15877987 TI - Noninvasive assessment of coronary plaque burden using multislice computed tomography. AB - We performed coronary plaque imaging with 16-row multislice computed tomography in 85 patients who had stable angina pectoris and a high pretest likelihood of having coronary plaque to evaluate plaque burden, i.e., extent (number of diseased coronary segments) and size (small vs large) of plaque. We also assessed type of plaque (calcified, noncalcified, or mixed) and its anatomic distribution. Of 85 patients included, 78 (92%) had fully evaluable multislice computed tomograms that allowed assessment of coronary plaque burden, including major and side branches (>or=2 mm), yielding a total of 855 segments. These 78 patients (92% men; mean age +/- SD 58 +/- 11.5 years) were in sinus rhythm, with heart rates of <70 beats/min (spontaneous or induced by beta blocker). Plaque was detected in 57% of all segments (487 of 855). The mean number of segments with plaque per patient +/- SD was 6.2 +/- 3.9. Plaque was classified as large in 33% of segments and small in 67%. Overall, 65% of plaques were calcified, 24% were noncalcified, and 11% were mixed. Plaques were predominantly located in the proximal and middle segments of the main coronary vessels. PMID- 15877988 TI - Relation of Basal coronary nitric oxide activity to the burden of atherosclerosis. AB - Coronary disease or its risk factors has been reported to attenuate basal nitric oxide (NO) activity. Intravascular ultrasound was used in the present study to better understand this relation. Basal and stimulated NO activities were assessed in 53 stable subjects. Coronary diameter and velocity (0.014-inch Doppler wire) were measured at baseline and after intracoronary infusion of the NO synthase inhibitor N(G)-monomethyl-l-arginine, acetylcholine (10(-6) M), nitroglycerin (200 microg), and adenosine (24 microg). Intimal thickening was quantified with intravascular ultrasound. N(G)-monomethyl-l-arginine resulted in significant decreases in coronary blood flow (-14 +/- 48%), proximal coronary diameter (-10 +/- 18%), and distal coronary diameter (-10 +/- 9%, all p values <0.0001). Basal NO activity was unrelated to the presence of coronary disease as assessed by angiography and the burden of atherosclerosis as assessed by intravascular ultrasound. Conversely, stimulated NO activity correlated inversely with burden of coronary atherosclerosis (p <0.05). Basal NO activity is relatively preserved in patients who have moderate coronary disease and is not related to the degree of atherosclerosis as assessed by intravascular ultrasound. This is in contradistinction to the impairment of stimulated NO activity in the coronary circulation that characterizes atherosclerosis. PMID- 15877989 TI - Association of metabolic syndrome with exercise capacity and heart rate recovery in patients with coronary heart disease in the heart and soul study. AB - It is not known whether the metabolic syndrome is associated with poor exercise capacity among patients who have established coronary heart disease. We evaluated the association of the metabolic syndrome with treadmill exercise capacity and heart rate recovery among patients who had coronary heart disease. We measured treadmill exercise capacity (METs) and heart rate recovery (beats per minute) in 943 subjects who had known coronary heart disease. Of these, 377 (40%) had the metabolic syndrome as defined by criteria of the National Cholesterol Education Program. Participants who had the metabolic syndrome were more likely to have poor exercise capacity (METs <5, 33% vs 18%, p <0.0001) and poor heart rate recovery (100 for subjects with metabolic syndrome were 1.7 (95% CI 1.3 to 2.3) for men and 1.6 (95% CI 1.2 to 2.1) for women. Associations were found for most of the components of the metabolic syndrome with CAC. Associations with the metabolic syndrome were similar for calcified atherosclerotic plaque in the abdominal aorta among 3,173 subjects, with adjusted odds ratios for a score >1,000 of 2.1 (95% CI 1.5 to 3.1) for men and 1.8 (95% CI 1.4 to 2.4) for women. We conclude that the metabolic syndrome and most of its components are associated with a higher prevalence of calcified atherosclerotic plaque in the coronary arteries and abdominal aorta in white and African-American men and women. PMID- 15877991 TI - Effect of lifestyle changes on atherogenic lipids and endothelial cell adhesion molecules in young adults with familial premature coronary heart disease. AB - Guidelines have recommended that a family history of premature coronary heart disease (CHD) warrants screening and preventive efforts, including lifestyle change. The objective of this study was to evaluate the effects of a lifestyle modification program on lipids and novel risk markers in young relatives of patients with premature CHD. In a parallel, randomized, intervention trial, intensified support to quit smoking and dietary modification was compared with general lifestyle advice in 172 men and women aged 18 to 39 years with a total cholesterol of 5 to 8 mmol/L and >or=1 lipid abnormality (high low-density lipoprotein [LDL] cholesterol, triglycerides or lipoprotein(a), or low high density lipoprotein cholesterol). All had a first-degree relative with premature CHD or with hyperlipidemia and other relative(s) with premature CHD, and 40% were daily smokers. After a mean of 8 months, the intervention group reduced the dietary intake of saturated fat and cholesterol compared with controls (p = <0.01). Ten smokers in the intervention group quit, whereas 2 subjects in the control group started and none quit. LDL cholesterol (p = 0.007), oxidized LDL (p = 0.03), and E-selectin (p = 0.02) concentrations were reduced in the intervention group compared with controls. In subjects who quit smoking, concentrations of intercellular adhesion molecule-1 decreased (261 +/- 66 to 228 +/- 30 ng/ml) compared with that in continuing smokers (308 +/- 106 ng/ml to 304 +/- 109 ng/ml) (p = 0.05 between groups). These findings indicate that dietary modification and smoking cessation are feasible in young adults with familial premature CHD and document novel mechanisms by which lifestyle modification may reduce CHD risk. PMID- 15877992 TI - Effect of resistance exercise on skeletal muscle myopathy in heart transplant recipients. AB - The purpose of this study was to determine the efficacy of resistance exercise in reversing skeletal muscle myopathy in heart transplant recipients. Myopathy, engendered by both heart failure and immunosuppression with glucocorticoids, is a post-transplant complication. The sequelae of myopathic disease includes fiber type shifts and deficits in aerobic metabolic capability. We randomly assigned patients to either 6 months of resistance exercise (training group; n = 8) or a control (control group; n = 7) group. Exercise was initiated at 2 months after transplant. Biopsy of the right vastus lateralis was performed before and after the 6-month intervention. Myosin heavy chain (MHC) composition was assessed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochemical assays were performed to determine citrate synthase, 3-hydroxyacyl-CoA-dehydrogenase, and lactate dehydrogenase activity. There were no group differences (p >or=0.05) in MHC composition and enzymatic reserve at baseline. Improvements in the training group for citrate cynthase (+40%), 3-hydroxyacyl-CoA-dehydrogenase (+10%), and lactate dehydrogenase activity (+48%) were significantly greater (p 200 mg) doses of aspirin (acetylsalicylic acid [ASA]) in 192,036 patients enrolled in 31 clinical trials. Despite substantial differences in the reporting patterns of bleeding complications, low-dose ASA was associated with the lowest risk, and moderate doses caused a relatively high hemorrhagic event rate, especially with regard to minor, gastrointestinal, and total bleeding, and stroke. These findings should be considered when using combination antiplatelets, anticoagulant therapy, or both, with ASA, especially with the daily dose of >100 mg. PMID- 15877995 TI - Myocardial technetium-99m-tetrofosmin single-photon emission computed tomography compared with 18F-fluorodeoxyglucose imaging to assess myocardial viability. AB - Technetium-99m-tetrofosmin single-photon emission computed tomography at rest is practical for routine assessment of viability in patients with ischemic cardiomyopathy. However, underestimation of viability may occur compared with 18F fluorodeoxyglucose single-photon emission computed tomography, especially in the inferoposterior and septal regions. PMID- 15877996 TI - Impact of coronary calcium score on diagnostic accuracy for the detection of significant coronary stenosis with multislice computed tomography angiography. AB - One hundred twenty patients in sinus rhythm with suspected coronary artery disease who underwent multislice computed tomography of the heart and conventional coronary angiography were retrospectively selected. The population was divided into 2 groups depending on their calcium score (CS) (e.g., low CS and high CS). The diagnostic accuracy of multislice computed tomographic scans for detecting significant lesions (>50% lumen reduction) in both groups was compared with quantitative coronary angiography. The sensitivity and specificity of multislice computed tomography were 90% and 92%, and 97% and 91% for low and high CS groups, respectively. PMID- 15877997 TI - Usefulness of chest pain character and location as diagnostic indicators of an acute coronary syndrome. AB - This study assessed chest pain locations/symptoms among patients who presented with acute chest pain and how these compare with a clinical diagnosis of cardiac or noncardiac chest pain. A cluster analysis was undertaken to determine any pattern in the chest pain locations described by patients. Cluster analysis identified 4 distinct chest pain locations (upper chest, central retrosternal, central chest, and left chest and left arm). There was considerable location/symptom overlap between patients who had cardiac chest pain and those who had noncardiac chest pain. PMID- 15877998 TI - Influence of reperfusion therapy on prognosis in patients aged >or=89 years with acute myocardial infarction. AB - There is no evidence on the efficacy of reperfusion therapies on nonagenarian patients who have acute myocardial infarction. The present study suggests that such therapies may not be useful and that thrombolysis may even be harmful. PMID- 15877999 TI - Effect of nifedipine on C-reactive protein levels in the coronary sinus and on coronary blood flow in response to acetylcholine in patients with stable angina pectoris having percutaneous coronary intervention. AB - The effects of nifedipine on inflammation and endothelial function in the coronary circulation were studied in patients who had angina pectoris (n = 17). Long-term treatment with nifedipine (nifedipine CR, 20 mg/day for 4 months) decreased levels of C-reactive protein in the coronary sinus (from 0.35 +/- 0.09 mg/dl to 0.07 +/- 0.01 mg/dl, mean +/- SEM, p <0.05) and enhanced acetylcholine induced increases in coronary blood flow. Thus, nifedipine is effective in decreasing inflammation and incresing endothelial function in the coronary circulation. PMID- 15878000 TI - Comparison of C-reactive protein levels before and after coronary stenting and restenosis among patients treated with sirolimus-eluting versus bare metal stents. AB - We prospectively studied the inflammatory response to coronary stenting (calculated as the difference between the highest postprocedural C-reactive protein [CRP] and baseline CRP levels [DeltaCRP]) and restenosis in 301 patients who received a sirolimus-eluting stent (n = 149) or a bare stent (n = 152) in the setting of a randomized trial. Median values of DeltaCRP were 3.1 mg/L in the sirolimus-eluting stent group and 3.0 mg/L in the bare stent group (p = 0.71). In the sirolimus-eluting group, restenotic rates were 9.7% in the subgroup with DeltaCRP higher than the median and 11.5% in the subgroup with DeltaCRP no higher than the median (p = 0.37). In the bare stent group, restenotic rates were 28.6% in the subgroup with DeltaCRP higher than the median and 15.4% in the subgroup with DeltaCRP no higher than the median (p = 0.04). PMID- 15878001 TI - Intravascular ultrasound stent area of sirolimus-eluting stents and its impact on late outcome. AB - Stent area assessed by intravascular ultrasound is a strong predictor of late outcome after percutaneous coronary intervention. Because sirolimus-eluting stents (SESs) prevent intimal regrowth within the stent, achieving a larger final stent area might no longer be noteworthy; therefore, the investigators sought to determine the relevance and impact of the final stent area with SESs. PMID- 15878002 TI - Meta-analysis of survival and relief of angina pectoris after transmyocardial revascularization. AB - A meta-analysis of 7 randomized trials involving 1,053 patients was performed to evaluate the effect of transmyocardial laser revascularization as the sole procedural intervention on survival and angina relief. At 1 year, transmyocardial laser revascularization produced a significant improvement in angina class (p <0.0001) but no improvement in survival (p = 0.75). PMID- 15878003 TI - Effect of low-density lipoprotein apheresis on lipoprotein-associated phospholipase A2. AB - Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is a proinflammatory participant in atherosclerosis and a potential biomarker for coronary heart disease. The effects of low-density lipoprotein (LDL) apheresis on Lp-PLA(2) levels were evaluated in 8 patients with cardiovascular disease. Each patient received 5 LDL apheresis treatments over a 3-month period. The mean direct LDL cholesterol level reduction was 60% (252 to 100 mg/dl). LDL apheresis acutely reduced Lp-PLA(2) by 21.4%. Over the course of treatment, Lp-PLA(2) levels were reduced by 29%. Chronic LDL apheresis significantly reduces Lp-PLA(2) independent of LDL cholesterol, which may be a potential mechanism by which LDL apheresis diminishes coronary heart disease risk. PMID- 15878004 TI - Influence of the randomized trials, AFFIRM and RACE, on the management of atrial fibrillation in two University Medical Centers. AB - The results of the Atrial Fibrillation Follow-Up Investigation of Rhythm Management (AFFIRM) and the Rate Control Versus Electrical Cardioversion for Persistent Atrial Fibrillation (RACE) study were presented in March 2002. These large studies showed no benefit of a rhythm-control strategy over a rate-control strategy in managing atrial fibrillation (AF). Cardioversion and atrioventricular junctional ablation are forms of rhythm control and rate control, respectively. The numbers of cardioversions and atrioventricular junctional ablations performed at the University of Virginia and the Medical College of Virginia during the 52 months before AFFIRM and RACE results were released and the 21 months afterward were compared. From January 1998 to March 2002, monthly averages of 31 +/- 8 elective cardioversions and 6 +/- 3 atrioventricular junctional ablations were performed; from April 2002 to December 2003, the monthly averages were 21 +/- 6 cardioversions (p = 0.001) and 9 +/- 3 ablations (p = 0.001). AF management changed at these institutions shortly after the RACE and AFFIRM results were released. PMID- 15878005 TI - Effect of conduction mode and location on electrophysiologic characteristics of accessory pathways. AB - The conduction properties of accessory pathways (APs) are independent of location and conduction mode (except in patients with multiple, Mahaim, and slowly conducting APs). Patients with right-sided APs show higher rates of atrial fibrillation and longer arrhythmia cycle length due to slower anterograde conduction over the atrioventricular node during atrioventricular reentrant tachycardia. PMID- 15878006 TI - Comparison of clinical presentation of acute myocarditis following smallpox vaccination to acute coronary syndromes in patients <40 years of age. AB - Smallpox vaccine-associated myopericarditis may have a similar presentation to acute coronary syndrome (ACS). The clinical records of 78 young patients (<40 years of age) presenting with ACS (n = 16) or myocarditis after smallpox vaccination (n = 62) were reviewed. Comparisons were made among clinical presentation, cardiac enzymes, echocardiographic findings, and electrocardiographic changes. The presence of cardiac risk factors or focal wall motion abnormalities on echocardiography were associated with a diagnosis of ACS. There was a trend toward earlier elevation of troponin-I and creatine kinase in patients with myocarditis compared with ACS. PMID- 15878007 TI - Effect of transcatheter atrial septal defect closure in children on left ventricular diastolic function. AB - The acute impact of transcatheter atrial septal defect (ASD) closure on left ventricular (LV) diastolic function was assessed in 15 consecutive children, and pre- and postclosure data were compared with those in a matched group of controls. The data indicate that transcatheter ASD closure leads to an immediate improvement in LV diastolic function as assessed by septal myocardial Doppler tissue imaging and LV inflow velocity propagation. The improvement in LV diastolic properties correlates with the degree of right ventricular volume overload. PMID- 15878008 TI - Correlation between apoptotic endothelial microparticles and serum interleukin-6 and C-reactive protein in healthy men. AB - Inflammation has been associated with increased cardiovascular risk, and endothelial cell (EC) apoptosis has been implicated in atherogenesis. The correlation between circulating concentrations of interleukin-6 (IL-6), C reactive protein (CRP), and endothelial microparticles (EMPs) expressing an apoptotic (EMP31) or activation (EMP62E) phenotype in 20 middle-aged healthy men was investigated. IL-6 was significantly correlated with EMP31 (r = 0.6, p = 0.004), which persisted after adjusting for body mass index and CRP. CRP was significantly correlated with body mass index (r = 0.49, p = 0.02) but not with EMP31 or EMP62E. EC apoptosis is associated with IL-6 levels in men and might be partially responsible for the increased cardiovascular risk associated with subclinical inflammation. PMID- 15878009 TI - Diagnosis of right ventricular dysfunction in acute pulmonary embolism using helical computed tomography. AB - Forty-six consecutive patients with pulmonary embolism (PE) who underwent pulmonary angiography, helical computed tomography (CT), and echocardiography in the investigators' emergency department were studied. It was determined that the CT right ventricular (RV)/left ventricular (LV) end-diastolic area ratio was correlated with PE obstruction and echocardiography. A CT RV/LV area ratio >1 had a sensitivity of 88% and a specificity of 88% in diagnosing significant PE. The present study suggests that helical CT may be used as a triage tool in acute PE for selecting high-risk patients, using calculation of the RV/LV area ratio to detect RV dysfunction. PMID- 15878010 TI - Comparison of accurate measurement of left ventricular mass in patients with hypertrophied hearts by real-time three-dimensional echocardiography versus magnetic resonance imaging. AB - To evaluate whether left ventricular (LV) mass assessed by a new real-time, 3 dimensional echocardiographic (RT-3DE) system corresponds to cardiac magnetic resonance imaging (MRI) in patients with LV hypertrophy, RT-3DE and 2-dimensional echocardiography (2DE) were performed to calculate LV mass in 21 patients (mean age 54 +/- 15 years) who underwent MRI for the evaluation of LV hypertrophy. In 20 of 21 patients, adequate 3-dimensional data for LV mass analysis were obtained, and regression analysis showed that LV mass by RT-3DE correlated with that determined by MRI (r = 0.95, y = 28.9 + 0.85x) better than with that determined by 2DE (r = 0.70, y = 43.6 + 0.81x). RT-3DE allows the accurate measurement of LV mass in patients with hypertrophied hearts. PMID- 15878011 TI - Ultrasonic biophysical measurements in the normal human fetus for optimal design of the monolithic fetal pacemaker. AB - Ultrasound measurements, including xiphoid-to-pericardial distance and deployment angle, were made on human fetuses as a function of gestational age for the purpose of assessing the likelihood of 3 failure modes of a monolithic fetal pacemaker, including primary positioning failure due to device length and secondary dislodgement failure due to somatic growth. The small variation of the measurements over the gestational age range relevant to device implantation for the major indications of the device (for complete heart block complicated by hydrops and for bradycardia risk after fetal surgery or intrauterine intervention) predicts a small likelihood of these failure modes. PMID- 15878012 TI - Effectiveness of echocardiographic imaging by nurses to identify left ventricular systolic dysfunction in high-risk patients. AB - Recognizing left ventricular (LV) systolic dysfunction is critical. The investigators sought to evaluate whether nurses could be trained to use a hand carried ultrasound (HCU) device to screen for LV systolic dysfunction in high risk patients. Sixty-three patients from an outpatient diabetes clinic underwent brief echocardiographic examinations by nurses using HCU devices. Of the 63 patients enrolled in the study, 3 (4.7%) had LV systolic dysfunction. The nurses correctly identified these 3 patients as having LV systolic dysfunction (sensitivity 100%, negative predictive value 100%). The identification of occult LV systolic dysfunction in diabetic patients may allow the initiation of therapies known to improve prognosis. PMID- 15878013 TI - Proceedings of the Editorial Board Meeting of the American Journal of Cardiology on 7 March 2005. PMID- 15878014 TI - Combined anaerobic-aerobic treatment of azo dyes--a short review of bioreactor studies. AB - The most logical concept for the removal of azo dyes in biological wastewater treatment systems is based on anaerobic treatment, for the reductive cleavage of the dyes' azo linkages, in combination with aerobic treatment, for the degradation of the products from azo dye cleavage, aromatic amines. Since the 1990s, several research papers have been published on combined, sequential or integrated, anaerobic-aerobic bioreactor treatment of azo dye-containing wastewater. The extent of azo dye reduction in the anaerobic phase of those bioreactor systems was generally high, albeit the process often required long reaction times, a limitation that can easily be remedied by making use of the property of redox mediators to speed up the process. The consequent removal of aromatic amines under aerobic conditions was less unequivocal. Although analytical data indicate that many of the aromatic amines were removed from the wastewater, and although the limited amount of available toxicity data all show far-reaching detoxification during aerobic treatment, it is clear that not all aromatic amines can be completely mineralized. PMID- 15878015 TI - Degradation of industrial waste waters on Fe/C-fabrics. Optimization of the solution parameters during reactor operation. AB - This study addresses the pre-treatment of toxic and recalcitrant compounds found in the waste waters arriving at a treating station for industrial effluents containing chlorinated aromatics and non-aromatic compounds, anilines, phenols, methyl-tert-butyl-ether (MTBE). By reducing the total organic carbon (TOC) of these waste waters the hydraulic load for the further bacterial processing in the secondary biological treatment is decreased. The TOC decrease and discoloration of the waste waters was observed only under light irradiation in the reactor by immobilized Fenton processes on Fe/C-fabrics but not in the dark. The energy of activation for the degradation of the waste waters was of 4.2 kcal/mol. The degradation of the waste waters was studied in the reactor as a function of (a) the amount of oxidant used (H2O2), (b) the recirculation rate, (c) the solution pH and (d) the applied temperature. With these parameters taken as input factors, statistical modeling allows one to estimate the most economic use of the oxidant and electrical energy to degrade these waste waters. The concentration of the most abundant organic pollutants during waste waters degradation was followed by gas chromatography/mass spectrometry (GC-MS). The ratio of the biological oxygen demand to the total organic carbon BOD5/TOC increased significantly due to the Fe/C-fabric catalyzed treatment from an initial value of 2.03 to 2.71 (2 h). The reactor results show that the recirculation rate has no influence on the TOC decrease of the treated waters but affects the BOD increase of these solutions. PMID- 15878016 TI - Influence of the sediment on lead speciation in the Tagus estuary. AB - The aim of this work is to study the influence of the Tagus estuarine sediment on lead speciation in the overlying natural water. The water sample in the presence of the sediment was contaminated three times with Pb(II) in a laboratory experiment. In different periods of time, at 1-7 days after each contamination, small volumes of water were titrated with lead. The titration was followed by anodic stripping voltammetry in differential pulse mode. Before and after contamination systematic analysis of the voltammetric parameters (peak current, peak potential and peak width) were carried out to get a clear picture of Pb(II) complexation in the soluble fraction in contact with the sediment. Two main types of organic ligands, macromolecular ligands and small compounds, were detected before contamination. Both of them form labile complexes (degree of lability within the timescale of some milliseconds). The small compounds, with a diffusion coefficient similar to that of the free metal ion, present a homogeneous behaviour in terms of Pb complexation. On the other hand macromolecular ligands, with a diffusion coefficient of 1.2 x 10(-6) cm2s(-1), can be described by two different binding groups, which might be of phenolic and carboxylic type as presented by humic matter. The sediment eliminated lead contamination (10(-6) moldm(-3) was the maximum concentration added) from 12 dm3 of water (surface of 8 dm2) within 2 days. It was also found that the sediment released organic ligands responsible for both labile and inert Pb complexes "seen" by voltammetry. The release of organic ligands that decreases the bioavailability of Pb(II) was clearly detected 1 week after contamination. Therefore, the sediment acts as a buffer for lead through two mechanisms against lead contamination: removing lead ions from the solution and releasing organic ligands to the water column. In a short period of time the sediment responds as a self-cleaning system for Pb(II) contamination in the estuarine water, which may have a very important influence in environmental pollution. PMID- 15878017 TI - Mineralisation of coloured aqueous solutions by ozonation in the presence of activated carbon. AB - The degradation of organic matter in coloured solutions of different classes of dyes by ozonation in the presence of activated carbon is investigated. The kinetics of the decolourisation and mineralisation of three different dyes solutions (CI Acid Blue 113, CI Reactive Red 241 and CI Basic Red 14) were studied in a laboratory scale reactor by three different processes: adsorption on activated carbon, oxidation with ozone and ozonation in the presence of activated carbon. The mineralisation of the solutions was followed by measuring the total organic carbon (TOC). Under the experimental conditions used in this work, activated carbon was not capable of completely removing the colour of the solutions in reasonable time. On the other hand, ozonation quickly decolourised all the solutions, but satisfactory removal of TOC was never achieved by this process. The combination of activated carbon with ozone enhanced the decolourisation of the solutions and especially the mineralisation of the organic matter. Activated carbon acts both as an adsorbent and as a catalyst in the reaction of ozonation. The surface chemistry of the activated carbon is an important parameter; it was observed that basic samples improve TOC removal. The main conclusions of this work were validated by treating a real textile effluent collected after the conventional biological treatment. PMID- 15878018 TI - Halophilic biological treatment of tannery soak liquor in a sequencing batch reactor. AB - Hypersaline wastewater (i.e. wastewater containing more than 35 gl(-1) total dissolved solids (TDS)) is generated by various industrial activities. This wastewater, rich in both organic matter and TDS, is difficult to treat using conventional biological wastewater treatment processes. Among the industries generating hypersaline effluents, tanneries are prominent in India. In this study, tannery wastewater from soak pit was treated in a lab-scale SBR for the removal of organic matter. The characterisation of the soak liquor showed that this effluent is biodegradable, though not easily, and highly variable, depending on the origin and the nature of the hides. TDS was in the range of 21-57 gl(-1) and COD was in the range of 1.5-3.6 gl(-1). This soak liquor was biologically treated in an aerobic sequencing batch reactor seeded with halophilic bacteria, and the performance of the system was evaluated under different operating conditions with changes in hydraulic retention time, organic loading rate and salt concentration. The changes in salinity appeared to affect the removal of organic matter more than the changes in hydraulic retention time or organic loading rate. Despite the variations in the characteristics of the soak liquor, the reactor achieved proper removal of organic matter, once the acclimation of the microorganisms was achieved. Optimum removal efficiencies of 95%, 93%, 96% and 92% on COD, PO4 3-, TKN and SS, respectively, could be reached with 5 days hydraulic retention time (HRT), an organic loading rate (OLR) of 0.6 kg COD m( 3)d(-1) and 34 g NaCl l(-1). The organisms responsible for nitrogen removal appeared to be the most sensitive to the modifications of these parameters. PMID- 15878019 TI - Strategies for changing temperature from mesophilic to thermophilic conditions in anaerobic CSTR reactors treating sewage sludge. AB - Thermophilic anaerobic digestion presents an advantageous way for stabilization of sludge from wastewater treatment plants. Two different strategies for changing operational process temperature from mesophilic (37 degrees C) to thermophilic (55 degrees C) were tested using two continuous flow stirred tank reactors operated at constant organic loading rate of 1.38 g VS/l reactor/day and hydraulic retention time of 20 days. In reactor A, the temperature was increased step-wise: 37 degrees C-->42 degrees C-->47 degrees C-->51 degrees C-->55 degrees C. While in reactor B, the temperature was changed in one-step, from 37 degrees C to the desired temperature of 55 degrees C, The results showed that the overall adaptation of the process for the step-wise temperature increment took 70 days in total and a new change was applied when the process was stabilized as indicated by stable methane production and low volatile fatty acids concentrations. Although the one-step temperature increase caused a severe disturbance in all the process parameters, the system reached a new stable operation after only 30 days indicating that this strategy is the best in changing from mesophilic to thermophilic operation in anaerobic digestion plants. PMID- 15878020 TI - Long-term simulation of the activated sludge process at the Hanover-Gummerwald pilot WWTP. AB - The aim of this study was to obtain a validated model, consisting of the Activated Sludge Model No. 3 (ASM3) and the EAWAG bio-P module, which could be used as a decision tool for estimating the maximum allowable peak flow to wastewater treatment plants during stormwater conditions. The databases used for simulations originated from the Hanover-Gummerwald pilot plant subjected to a series of controlled, short-term hydraulic shock loading experiments. The continuous influent wastewater composition was generated using on-line measurements of only three parameters (COD, N-NH4+, P-PO4 3-). Model predictions were compared with on-line data from different locations in the activated sludge system including the aerobic zone (concentrations of N-NH4+, N-NO3-) and secondary effluent (concentrations of P-PO4 3-). The simulations confirmed experimental results concerning the capabilities of the system for handling increased flows during stormwater events. No (or minor) peaks of N-NH4+ were predicted for the line with the double dry weather flowrate, whereas peaks of N NH4+ at the line with the quadruple dry weather flowrate were normally exceeding 8 g Nm(-3) (similar to the observations). PMID- 15878021 TI - Denitrification on poly-beta-hydroxybutyrate in microbial granular sludge sequencing batch reactor. AB - Microbial granules were successfully cultivated in an alternating aerobic anaerobic sequencing batch reactor (SBR) for removing organic carbon and nitrogen. It was found that almost all input ammonium was converted to nitrite and nitrate in the aerobic phase, while the efficiency of denitrification was highly related to the availability of external carbon source in the anaerobic phase. Complete denitrification was achieved with sufficient supply of external carbon, while only partial denitrification was observed with no addition of external carbon. Results showed that in the absence of external carbon source, pre-accumulated poly-beta-hydroxybutyric acid (PHB) in microbial granules could be utilized for cell maintenance and denitrification. With supply of external carbon but no addition of nitrate, PHB accumulation accounted for the main mechanism of the organic removal. Under balanced growth conditions (with organic carbon and nitrogen supply), external organic carbon was consumed simultaneously for denitrification, PHB storage as well as for cell functions. It was revealed that the potential role of PHB for denitrification by microbial granules was very limited, i.e. less than 28 mg nitrate-nitrogen l(-1) was found to be denitrified with internally accumulated PHB. This study for the first time shows the limiting capacity of PHB as reducing power for denitrification by microbial granules. PMID- 15878022 TI - Optimum cycle time for intermittent UASB reactors treating dairy wastewater. AB - This work accesses the influence of cycle duration on the intermittent operation of mesophilic UASB reactors inoculated with flocculent sludge and used in dairy wastewater (DWW) treatment. Five cycle lengths ranging from 24 to 144 h were compared for loads between 2.5 and 29.0 gCOD/l/d. COD balances are presented which demonstrate the importance of a feedless period in the conversion to methane of the substrates removed during the feed period. The maximum applicable loads determined for the system were higher for the longer cycle times. The higher conversion to methane of the removed COD was obtained for the 96 h cycle (48 h feed + 48 h feedless) resulting in a more stable operation. The 96 h cycle (48 h feed + 48 h feedless) was considered as the optimum for the treatment of dairy effluents in intermittent UASB reactors. Compared to the maximum applicable loads reported in the literature for continuous systems (3-6 gCOD/l/d) treating dairy effluents the stable operation loads attained with intermittent operation were considerably higher (22 gCOD/l/d). PMID- 15878023 TI - Inactivation of enteric microorganisms with chemical disinfectants, UV irradiation and combined chemical/UV treatments. AB - The relative disinfection efficiencies of peracetic acid (PAA), hydrogen peroxide (H2O2) and sodium hypochlorite (NaOCl) against Escherichia coli, Enterococcus faecalis, Salmonella enteritidis and coliphage MS2 virus were studied in laboratory-scale experiments. This study also evaluated the efficiency of combined PAA/ultraviolet irradiation (UV) and H2O2/UV treatments to determine if the microbial inactivation was synergistic. Microbial cultures were added into a synthetic wastewater-like test medium and treated by chemical disinfectants with a 10 min contact time, UV irradiation or the combination of chemical and UV treatments. A peracetic acid dose of 3 mg/l resulted in approximately 2-3 log enteric bacterial reductions, whereas 7-15 mg/l PAA was needed to achieve 1-1.5 log coliphage MS2 reductions. Doses of 3-150 mg/l hydrogen peroxide achieved below 0.2 log microbial reductions. Sodium hypochlorite treatments caused 0.3-1 log microbial reductions at an 18 mg/l chlorine dose, while 2.6 log reductions of E. faecalis were achieved at a 12 mg/l chlorine dose. The results indicate that PAA could represent a good alternative to chlorine compounds in disinfection procedures, especially in wastewaters containing easily oxidizable organic matter. Hydrogen peroxide is not an efficient disinfectant against enteric microorganisms in wastewaters. The combined PAA/UV disinfection showed increased disinfection efficiency and synergistic benefits with all the enteric bacteria tested but lower synergies for the coliphage MS2. This suggests that this method could improve the efficiency and reliability of disinfection in wastewater treatment plants. The combined H2O2/UV disinfection only slightly influenced the microbial reductions compared to UV treatments and showed some antagonism and no synergies. PMID- 15878024 TI - Influence of chemical conditioning on the ion exchange capacity and on kinetic of zinc uptake by clinoptilolite. AB - Aim of this study was to evaluate the feasibility of the use of clinoptilolite as a barrier material to eliminate heavy metals from roof runoff. The effect of chemical conditioning with 1 M NaCl solution upon the ion exchange capacity and on kinetic of zinc uptake by clinoptilolite has been investigated. According to the batch experiments the modified clinoptilolite has up to 100% higher sorption capacity, regarding Zn than the natural material. The pre-treatment of clinoptilolite results in an acceleration of the ion exchange process up to 40% regarding zinc. In order to define the reasons of this behaviour, both materials, modified and natural, were analysed for: (i) chemical composition, (ii) density, (iii) pore size distribution and (iv) zeta potential. The clogging of the pores, the charge of the grain surface, the pH of the initial metal solution and the ion metal concentration are the factors which are mainly affecting the ion exchange capacity and the rate of zinc uptake by clinoptilolite. PMID- 15878025 TI - Evaluation of empirical formulae for estimation of the longitudinal dispersion in activated sludge reactors. AB - Tracer studies are widely applied to characterize the hydraulic properties of reactors. In the case of activated sludge reactors, however, tracer test results are difficult to interpret due to internal and returned activated sludge recirculation. Empirical formulae can be considered as an alternative method of estimating the hydraulic conditions within the activated sludge reactor. The aim of this study is to evaluate accuracy of four empirical formulae for the full scale conditions based on the results of tracer studies performed at the Rock Creek Wastewater Treatment Plant (WWTP) in Hillsboro, OR (USA). Values of the dispersion coefficient, E(L), were first estimated using a 1-D advection dispersion equation and setting a sum of squares of differences between the observed and calculated tracer concentrations to a minimum. The estimated values of E(L) coefficient remained within the range of 1043-1580 m2/h. The best approximation of dispersion was obtained from the formula of Fujie et al. (1983, J. Ferment. Technol. 63(3), 295). Also the formula of Murphy and Boyko (1970, J. San. Eng. ASCE 96(2), 211) generated E(L) values of the same order as the optimum E(L). The accuracy of these formulae was further confirmed based on the results of studies reported in the literature. PMID- 15878026 TI - Anaerobic digestion of the organic fraction of municipal solid waste: influence of co-digestion with manure. AB - Anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) was investigated in two thermophilic (55 degrees C) wet digestion treatment systems R1 and R2. Initially OFMSW was co-digested with manure with a successively higher concentration of OFMSW, at a hydraulic retention time (HRT) of 14-18 d and an organic loading rate (OLR) of 3.3-4.0 g-VS/l/d. Adaptation of the co-digestion process to a OFMSW:manure ratio of 50% (VS/VS) was established over a period of 6 weeks. This co-digestion ratio was maintained in reactor R2 while the ratio of OFMSW to manure was slowly increased to 100% in reactor R1 over a period of 8 weeks. Use of recirculated process liquid to adjust the organic loading to R1 was found to have a beneficial stabilization effect. The pH rose to a value of 8 and the reactor showed stable performance with high biogas yield and low VFA levels. The biogas yield from source-sorted OFMSW was 0.63-0.71 l/g-VS both in the co digestion configuration and in the treatment of 100% OFMSW with process liquid recirculation. This yield is corresponding to 180-220 m3 biogas per ton OFMSW. VS reduction of 69-74% was achieved when treating 100% OFMSW. None of the processes showed signs of inhibition at the free ammonia concentration of 0.45-0.62 g-N/l. PMID- 15878027 TI - Water surface contacting cover system--the basic study for improving the oxygen transfer coefficient and the BOD removal capacity. AB - In order to construct an environmental aeration basin in a sewage treatment plant, the suggested novel aeration basin was examined for the oxygen transfer coefficient by using tap water on a lab-scale and pilot-scale. We called it water surface contacting cover system (WSCCS) that was installed with a cover plate on the wafer surface and a slant plate in aeration basin, compared to a standard aeration basin (SRAS, spiral roll aeration system). Finally, the novel basin could be compared by adopting the slant and cover plates into the actual sewage treatment plant (W x L x D (m)=5.0 x 2.65 x 4.25, treatment capacity: 600 m3/d). As a result, only by adding a slant plate, the oxygen transfer coefficient (K(L)a) increased approximately by 15% more than that with a SRAS basin. And the slope of slant plate was optimized at 10 degrees . Additionally, in the WSCCS, the increasing efficiency of the oxygen transfer coefficient, affected only by the cover, was proved at about 25% on the lab-scale aeration basin and about 20% on the pilot plant. After all, the increasing effect of the oxygen transfer coefficient on WSCCS that was installed with the water contacting cover and the slant plate on standard aeration basin was proved to be above 35%. The distance between the air diffuser and the slant plate was 20-30 cm in the pilot-scale plant. As an effective factor for increasing the oxygen transfer rate, the width/depth ratio of aeration basin was proved to be 2. Through the short-term results of actual sewage treatment plant, it was known that BOD removal efficiency could be improved by about 3% by adopting tile WSCCS not but SRAS. PMID- 15878028 TI - Degradation of atrazine in a laboratory scale model system with Danube river sediment. AB - A laboratory-scale model system of Danube river gravel bed was constructed. Evaluation of water chemical parameters and composition of bacterial communities confirmed the liability of the model. Degradation of atrazine, a frequent micropollutant of Danube water was examined in the model system. In case of an acute atrazine load, short-term retention of the compound was observed, accumulation and degradation were not significant. Long-term continuous dose of atrazine led to enhanced degradation of the pollutant. Principal metabolite was hydroxyatrazine. Atrazine utilizing strains isolated from the sediment belong to various, predominantly Gram positive genera and have diverse atrazine metabolism. Dominant metabolic reactions are dechlorination, dealkylation and deamination, with the sequential products ammeline and ammelide. The strain Delftia acidovorans D24 mineralizes atrazine as a sole source of carbon and nitrogen. PMID- 15878029 TI - Enhancement of anaerobic batch digestion of sisal pulp waste by mesophilic aerobic pre-treatment. AB - Pre-treatment of sisal pulp prior to its anaerobic digestion was investigated using an activated sludge mixed culture under aerobic conditions in batch bioreactors at 37 degrees C. The progression of aerobic pre-treatment of the residue in relation to the activities of some extracellular hydrolytic enzymes in the slurry was monitored. The highest activity of hydrolytic enzymes was obtained at 9 h of pre-treatment. Filter paper cellulase had a maximum activity of 0.90 IU/ml, while carboxymethyl cellulase, amylase and xylanase were produced to a maximum of about 0.40 IU/ml. The methane yield obtained after anaerobic digestion of the pre-treated pulp ranged between 0.12 and 0.24 m3 CH4/kg VS added. The highest and lowest values were obtained for 9 and 72 h of pre-treatment, respectively. Nine hours of pre-treatment of sisal pulp prior to anaerobic digestion demonstrated a 26% higher methane yield when compared to the sisal pulp without pre-treatment. The consortia of microorganisms in activated sludge demonstrated a useful potential in the production of hydrolases acting on major macromolecules of sisal pulp. The fact that a correlation was observed between high enzyme activity and high methane yield at 9 h of aerobic pre-treatment suggests that such a short pre-treatment period could be an alternative option for increasing solubilization of sisal pulp and promoting methane productivity. PMID- 15878030 TI - Effect of wastewater composition on archaeal population diversity. AB - Distribution and occurrence of Archaea and methanogenic activity in a laboratory scale, completely mixed anaerobic reactor treating pharmaceutical wastewaters were investigated and associated with reactor performance. The reactor was initially seeded with anaerobic digester sludge from an alcohol distillery wastewater treatment plant and was subjected to a three step feeding strategy. The feeding procedure involved gradual transition from a glucose containing feed to a solvent stripped pharmaceutical wastewater and then raw pharmaceutical wastewater. During the start-up period, over 90% COD removal efficiency at an organic loading rate (OLR) of 6 kg COD m(-3)d(-1) was achieved with glucose feeding, and acetoclastic methanogenic activity was 336 ml CH4 gTVS(-1)d(-1). At the end of the primary loading, when the feed contained solvent stripped pharmaceutical wastewater at full composition, 71% soluble COD removal efficiency was obtained and acetoclastic methanogenic activity decreased to half of the rate under glucose feed (166 ml CH4 gTVS(-1)d(-1)). At the end of secondary loading with 60% (w/v) raw pharmaceutical wastewater, COD removal dropped to zero and acetoclastic methanogenic activity fell to less than 10 ml CH4 gTVS(-1)d(-1). Throughout the course of the experiment, microbial community structure was monitored by DGGE analysis of 16S rRNA gene fragments. Five different archaeal taxa were identified and the predominant archaeal sequences belonged to methanogenic Archaea. Two of these showed greatest sequence identity with Methanobacterium formicicum and Methanosaeta concilii. The types of Archaea present changed little in response to changing feed composition but the relative contribution of different organisms identified in the archaeal DGGE profiles did change. PMID- 15878031 TI - A rapid method to quantify nitrifiers in activated sludge. AB - Quantification of bacteria using Fluorescence In Situ Hybridization (FISH), confocal laser scanning microscopy (CLSM) and image analysis is very time consuming and requires the availability of an expensive microscope. Therefore, a rapid method to quantify nitrifying bacteria in activated sludge using FISH and epifluorescence microscopy was developed. The quantification of the biovolume is based on manual counting of the aggregates formed by nitrifying bacteria and determination of their size. The overall uncertainty of the method was evaluated as a function of the number of analyzed microscopic fields. It was found that 10 15 microscopic fields for ammonia-oxidizing bacteria and 6-8 microscopic fields for nitrite-oxidizing bacteria per sample were optimal regarding effort and accuracy. Accordingly, the time needed for one sample was only 5-15 min, compared to about 1h for the quantification with CLSM and image analysis. As a consequence, this method also allows for the measurement of extended time series with a reasonable effort. The comparison of the determined biovolume and the measured activity showed an explicit correlation. PMID- 15878032 TI - Dichloroaniline retention by nanofiltration membranes. AB - This study evaluates the performance of two nanofiltration membranes in removing a herbicide: dichloroaniline. The membranes, one polyamide and one cellulose acetate, have a cut-off in the range 150-300 g/mol (manufacturers' data). The experiments were carried out with solutions of dichloroaniline in demineralized water, with concentrations from 1 to 10 ppb. For each membrane, the amount of herbicide retained and adsorbed by the membrane was determined as a function of feed concentration and transmembrane pressure. The two membranes, made of different materials but having the same nominal cut-off, retained dichloroaniline to very different extents and by different mechanisms. PMID- 15878033 TI - Influence of anode material on electrochemical oxidation for the treatment of tannery wastewater. AB - The treatment of tannery wastewater by electrochemical oxidation, mediated by an electro-generated species was carried out under galvanostatic conditions in an electrochemical reactor equipped with anodes based on noble metals and metal oxides (Ti/Pt-Ir, Ti/PbO2, Ti/PdO-Co3O4 and Ti/RhO(x)-TiO2). The decrease in time of chemical oxygen demand, nitrogen (TKN and ammonia), Cr and sulphides was monitored. The study showed that the rate of pollutant removal was significantly influenced by the type of anode material and electrochemical parameters. Different mechanisms contributed to the removal of pollutants when the reactor operated under conditions close to the limiting current for chlorine evolution and under much higher current density, with the reactor performing better at a high current/voltage. The kinetic pseudo-first order model applied for the interpretation of the results showed that the Ti/Pt-Ir and Ti/PdO-Co3O4 anodes performed better than the other two electrodes under the majority of tested conditions, with the highest rate of removal obtained for ammonia (kinetic rate constant k=0.75 min(-1)). Electrochemical oxidation can be applied as a post treatment after the conventional biological process in order to remove the residual ammonia with low energy consumption (0.4 kWh m(-3)). PMID- 15878034 TI - Anaerobic biological treatment of phenolic wastewater at 15-18 degrees C. AB - Low-temperature, or psychrophilic (<20 degrees C) anaerobic digestion has been proven feasible for the mineralisation of simple wastewaters. In this study, hybrid expanded granular sludge bed-anaerobic filter (EGSB-AF) bioreactors were used to evaluate the feasibility of psychrophilic digestion for the treatment of phenol-containing wastewater. Efficient chemical oxygen demand and phenol removal were observed at organic and phenol loading rates of 5 kg COD m(-3)d(-1) and 0.4 1.2 kg phenol m(-3)d(-1) (400-1200 mg phenol [l wastewater](-1)), respectively. There was no long-term accumulation of volatile fatty acids in the reactor systems. Methanogenic activity was developed under psychrophilic conditions but anaerobic methane-producing populations remained mesophilic throughout the trial of 415 days. PMID- 15878035 TI - Enumeration of bifidobacterial populations with selective media to determine the source of waterborne fecal pollution. AB - Bifidobacteria are one of the main groups of bacteria found in the intestine of humans and other animals. They have been proposed as indicators to identify the source of fecal pollution, since certain Bifidobacterium spp. are found only in humans. However, species identification remains difficult, and has limited their practical application. In this study, a simple assay based on the ratio of sorbitol-fermenting bifidobacteria to total colony forming units on Human Bifido Sorbitol Agar (HBSA) medium was developed. Ratios greater than 0.2 were indicative of human fecal pollution while animal-derived slaughterhouse effluent had a ratio less than 0.05. This is a low-cost, easily implemented approach which may have wide applicability when characterizing recent and high concentrated fecal pollution. PMID- 15878036 TI - Kinetics of reactions between chlorine and the cyanobacterial toxins microcystins. AB - Blooms of cyanobacteria can give rise to the production of toxins which contaminate drinking water sources. Among the oxidants and disinfectants typically applied in waterworks, chlorine has been found to be effective for the degradation of microcystins. In the present study, unknown second-order rate constants for the reactions of microcystin-LR (MC-LR), -RR and -YR with chlorine were determined over a wide pH range. It was found that an increase of pH has a negative effect on the microcystin degradation rate. Apparent second-order rate constant for the chlorination of MC-LR at 20 degrees C varied from 475 M(-1)s(-1) at pH 4.8 to 9.8 M(-1)s(-1) at pH 8.8. From these apparent second-order rate constants, rate constants for the reactions of MC-LR with hypochlorous acid (HOCl) and hypochlorite (ClO-) were evaluated. Half-life times ranged from minutes at pH 6 to 1 h at pH 8 for a constant residual chlorine concentration of 1.0-0.5 mgl(-1), typical of oxidation pre-treatment and final disinfection. Similar reactivity with chlorine was found for MC-RR and MC-YR. Therefore, chlorination is a feasible option for microcystin degradation during oxidation and disinfection processes, and can be applied in drinking water treatment in case of cyanobacterial toxin risk if the pH is kept below 8. PMID- 15878037 TI - Biosorption of protons and heavy metals onto olive pomace: modelling of competition effects. AB - Heavy metal biosorption onto solid wastes from olive oil production plants, olive pomace, has been investigated. Acid-base properties of the active sites of olive pomace were determined by potentiometric titrations and represented by a continuous model accounting for two main kinds of active sites. Competition among protons and heavy metals in solution was considered by performing biosorption tests at different equilibrium pH with single (Cu and Cd) and binary metal systems (Cu-Cd). Both Langmuir extensions and non-ideal competitive adsorption models (NICA models) can be used to represent experimental data of Cu and Cd biosorption in single metal systems at different equilibrium pH. Nevertheless only NICA models, accounting for site heterogeneity and non-ideal adsorption of the different species simultaneously present in solution, can adequately simulate the competition among Cu and Cd in binary metal systems by using the parameters fitted to single system data. PMID- 15878038 TI - Heterogeneous catalytic degradation of cyanide using copper-impregnated pumice and hydrogen peroxide. AB - The main objective of this research was to investigate the oxidative destruction of free cyanide with hydrogen peroxide and copper-impregnated pumice as a heterogeneous catalyst. Original or copper-impregnated pumices added alone were not effective adsorbents of negatively charged cyanide ions due to incompatible surface interactions. Peroxide and original pumices added together were also ineffective in removing cyanide. However, for all of the three natural pumices tested with various particle size fractions, the use of copper-impregnated pumices and peroxide together significantly enhanced both the initial rate and extent of cyanide removal. Although copper-impregnated specific surface area was the major factor affecting the rate and extent of cyanide destruction for a particular pumice source with similar surface chemistries, the type of surface chemistry (i.e., specific functional groups) within different pumice sources also appears to be a very important factor. Lower rates and extents of cyanide removals were observed at pH 11 compared to pH 8 probably because of the negative impacts of alkaline conditions in terms of scavenging peroxide and forming more negatively charged pumice surfaces. Both the initial rate and ultimate extent of cyanide removals were generally higher at a temperature of 20 degrees C compared with those found at 10 degrees C. The use of copper-impregnated pumice as a light, cheap, readily available, natural, and porous heterogeneous catalyst either in completely mixed/suspended or fixed-bed reactor configurations may be an effective treatment technology for cyanide removal from solution. This new approach may minimize downstream metal removal problems experienced in conventional cyanide oxidation technologies. PMID- 15878039 TI - Effects of activated carbon characteristics on the simultaneous adsorption of aqueous organic micropollutants and natural organic matter. AB - The overall objective of this research was to determine the effects of physical and chemical activated carbon characteristics on the simultaneous adsorption of trace organic contaminants and natural organic matter (NOM). A matrix of 12 activated carbon fibers (ACFs) with three activation levels and four surface chemistry levels (acid-washed, oxidized, hydrogen-treated, and ammonia-treated) was studied to systematically evaluate pore structure and surface chemistry phenomena. Also, three commercially available granular activated carbons (GACs) were tested. The relatively hydrophilic fuel additive methyl tertiary-butyl ether (MTBE) and the relatively hydrophobic solvent trichloroethene (TCE) served as micropollutant probes. A comparison of adsorption isotherm data collected in the presence and absence of NOM showed that percent reductions of single-solute TCE and MTBE adsorption capacities that resulted from the presence of co-adsorbing NOM were not strongly affected by the chemical characteristics of activated carbons. However, hydrophobic carbons were more effective adsorbents for both TCE and MTBE than hydrophilic carbons because enhanced water adsorption on the latter interfered with the adsorption of micropollutants from solutions containing NOM. With respect to pore structure, activated carbons should exhibit a large volume of micropores with widths that are about 1.5 times the kinetic diameter of the target adsorbate. Furthermore, an effective adsorbent should possess a micropore size distribution that extends to widths that are approximately twice the kinetic diameter of the target adsorbate to prevent pore blockage/constriction as a result of NOM adsorption. PMID- 15878041 TI - From home to street: understanding young people's transitions into homelessness. AB - This paper explores why young people leave home and become homeless. Drawing on life history interviews conducted with 50 homeless youth in Los Angeles, explanations provided by participants for becoming homelessness and how they understand their experiences are presented. In professional discourses, homeless young people are often portrayed as victims of physical abuse and emotional neglect. Although participants' narratives reveal that abuse and neglect play a central role in their decisions to leave home, many maintain a sense of agency in the recounting of how they became homeless. The importance of service providers recognizing young people's agency is discussed. PMID- 15878042 TI - Young people, drug use and family conflict: pathways into homelessness. AB - Young people who experience homelessness, in Australia and in other western contexts (US, Canada, England), are widely perceived to use and abuse alcohol and drugs. The available research indicates that homeless young people use all drug types, whether injected or otherwise, more frequently than their home-based peers. Debate exists in the research and policy literature about whether drug use is a cause or consequence of homelessness. In a study exploring homeless young peoples reasons for leaving home, we examined the relationship between young people's drug use and their pathways into homelessness. Brief qualitative interviews were conducted with 302 homeless young people (12-20 years). Following a thematic analysis of interview transcripts, four pathways into homelessness involving personal or familial drug use were identified. One-third of the participants indicated that personal or familial drug use was a critical factor in them leaving home. Of these, just over half indicated that personal drug use was a direct or indirect cause of their homelessness and one-quarter indicated that familial drug and alcohol use was the critical factor that led them to leaving home. One-quarter indicated that their drug use only began after they became homeless. Family conflict, if not family breakdown, was implicated in all four pathways out of home. PMID- 15878043 TI - Homelessness and drug abuse among young men who have sex with men in New York city: a preliminary epidemiological trajectory. AB - The objective of this paper is to profile the role of homelessness in drug and sexual risk in a population of young men who have sex with men (YMSM). Data are from a cross-sectional survey collected between 2000 and 2001 in New York City (N = 569). With the goal of examining the import of homelessness in increased risk for the onset of drug and sexual risk, we compare and contrast three subgroups: (1) YMSM with no history of homelessness, (2) YMSM with a past history of homelessness but who were not homeless at the time of the interview, and (3) YMSM who were currently homeless. For each group, we describe the prevalence of a broad range of stressful life events (including foster care and runaway episodes, involvement in the criminal justice system, etc.), as well as selected mental health problems (including past suicide attempts, current depression, and selected help-seeking variables). Additionally, we examine the prevalence of selected drug and sexual risk, including exposure to a broad range of illegal substances, current use of illegal drugs, and prevalence of lifetime exposure to sex work. Finally, we use an event history analysis approach (time-event displays and paired t-test analysis) to examine the timing of negative life experiences and homelessness relative to the onset of drug and sexual risk. High levels of background negative life experiences and manifest mental health distress are seen in all three groups. Both a prior experience of homelessness and currently being homeless are both strongly associated with both higher levels of lifetime exposure to drug and sexual risk as well as higher levels of current drug and sexual risk. Onset of these risks occur earlier in both groups that have had an experience of housing instability (e.g., runaway, foster care, etc.) but are delayed or not present among YMSM with no history of housing instability. Few YMSM had used drug prior to becoming homeless. While causal inferences are subject to the limitations of a cross-sectional design, the findings pose an empirical challenge to the prevailing assumption that prior drug use is a dominant causal factor in YMSM becoming homeless. More broadly, the data illustrate the complexity of factors that must be accounted for, both in advancing our epidemiological understanding of the complexity of homelessness and its relationship to the onset of drug and sexual risk among high risk youth populations. PMID- 15878044 TI - Factors associated with homelessness of adolescents under supervision of the youth protection system. AB - There are two factors that limit our knowledge of the risk factors associated with homelessness among runaway adolescents, namely (1) the samples used are often composed of youth homeless service users and/or youths living on the streets (visible homelessness), whereas most adolescents in fact use "private" resources (hidden homelessness), and (2) failure to use an adequate control group to identify risk factors associated specifically with homelessness. Our study compares the characteristics of two groups of youths under the supervision of the youth protection system, according to the presence or absence of periods of homelessness. The results throw light on the factors underlying the shift from "at risk" to "homeless", showing that youths with experience of homelessness are more likely to have been placed in substitute home environments, have experienced significant relationship difficulties with one of their parents (deterioration of the parent/youth relationship and parental abuse) and to have been diagnosed with behavioural disorders. The findings suggest that the decision to place young people under supervision is based more on the dynamic between risk factors rather than on the existence of behavioural problems. PMID- 15878045 TI - Predictors of social network composition among homeless and runaway adolescents. AB - Recent research on the social support networks of homeless and runaway youth suggest the social networks of runaway youth are made up largely of transient deviant peer relationships. This paper examined social network characteristics of 428 homeless and runaway adolescents from small-to moderate-sized cities in four Midwestern states. We investigated size, homogeneity, and correlates of the composition of the instrumental and emotional support networks as reported by the adolescents. Results showed the networks are considerably heterogeneous, comprised of relationships from home and the street as well as family and non related adults. Further, the composition of these networks is related to adolescent characteristics and experiences including sexual identity, abuse history, and street experience. PMID- 15878046 TI - Future expectations of Brasilian street youth. AB - Future expectations of youth surviving on the streets of Porto Alegre, Brasil, were examined. The sample consisted of 35 boys and 34 girls aged 10-18 (M age 14.4) who participated in a sentence completion task and semi-structured interviews. Responses to two incomplete sentences regarding the future revealed a mismatch between hoped-for and expected events. When completing the sentence, "In the future, I hope..." the majority of youth gave optimistic (but generally non specific) responses. In contrast, the sentence "For me, the future..." elicited more pessimistic responses. Few gender or age-related differences emerged. Findings are discussed in light of prior research with general populations of adolescents as well as studies of street youth. Taken as a whole, results suggest that life on the street shapes and constrains how youth see their futures. Implications of the findings for policy makers and practitioners are addressed. PMID- 15878047 TI - Predictors of close family relationships over one year among homeless young people. AB - Predictors of perceived family bonds were examined among homeless young people who initially left home one year earlier. Newly homeless young people aged 12-20 years who had recently left home were recruited in Los Angeles County, United States (n = 201) and Melbourne, Australia (n = 124) and followed longitudinally at 3, 6, and 12 months (follow-up rates ranging from 72% to 86% overall). These homeless young people varied substantially in their bonds to their families. Family bonds at one year were predicted in multivariate regression analyses by having significantly fewer problem behaviours when leaving home and decreasing rates of problem behaviours over the next year. Having more emotional support and more instrumental financial support were also significantly associated with greater family bonds one year later. These results suggest that efforts to reunite families may be a viable intervention strategy for newly homeless young people. PMID- 15878048 TI - Ecologically based family therapy outcome with substance abusing runaway adolescents. AB - Runaway youth report a broader range and higher severity of substance-related, mental health and family problems relative to non-runaway youth. Most studies to date have collected self-report data on the family and social history; virtually no research has examined treatment effectiveness with this population. This study is a treatment development project in which 124 runaway youth were randomly assigned to (1) ecologically based family therapy (EBFT) or (2) service as usual (SAU) through a shelter. Youth completed an intake, posttreatment, 6 and 12 months follow-up assessment. Youth assigned to EBFT reported greater reductions in overall substance abuse compared to youth assigned to SAU while other problem areas improved in both conditions. Findings suggest that EBFT is an efficacious intervention for this relatively severe population of youth. PMID- 15878049 TI - Microbial contamination of immersion biometry ultrasound equipment. AB - PURPOSE: To investigate the prevalence of microorganisms on ultrasound biometry equipment and cleaning habits. DESIGN: Observational case series. PARTICIPANTS: Thirty-four university-based and private ophthalmology clinics. METHODS: In a prospective multicenter study, clinics representative of every region of the country sampled their fixed immersion biometry equipment (i.e., ultrasound probe, immersion shell, and infusion tubing) for bacteria and fungi. Assessment of the cleaning habits for this equipment was conducted by way of a standard questionnaire that included type of fluid and delivery method, frequency of fluid change, method of cleaning the probe and shell, and frequency of tubing change and/or cleaning. MAIN OUTCOME MEASURES: Frequency (prevalence), descriptive statistics, and type of microorganisms. RESULTS: Eighteen samples (53% [18/34]) grew organisms from either the probe/shell or tubing. Positive cultures were found in 32% (11/34) of the immersion shell/probes and in 31% (10/32) of the infusion tubing samples. The bacteria most commonly cultured from both probe/shell and tubing was coagulase-negative Staphylococcus, whereas Penicillium species was the most commonly cultured fungus (exclusively from the probe/shell). Overall, fungi (Penicillium and Alternaria species) were cultured in 12% of the probe/shell samples. Only 14% of the study sites adequately disinfected the probe/shell according to Centers for Disease Control and Prevention (CDC) guidelines, which recommend a 5-minute soak in antiseptic. CONCLUSIONS: The bacteria and fungi that colonize biometry equipment are not being adequately eliminated by the cleaning/disinfecting techniques employed in most ophthalmology clinics. These results also may apply to contact biometry, pachymetry, and tonometry equipment as well. Clinicians should follow the CDC recommendations for disinfecting instruments that come in contact with the eye, and the infusion tubing should be changed after each patient. PMID- 15878050 TI - Discoid lupus erythematosus masquerading as chronic blepharoconjunctivitis. AB - PURPOSE: To recognize discoid lupus erythematosus (DLE) as a treatable cause of chronic blepharoconjunctivitis. DESIGN: Retrospective observational case series. PARTICIPANTS: Records of 5 patients with biopsy-proven DLE were reviewed. METHODS: Clinical and pathology records were examined. MAIN OUTCOME MEASURES: Patients' clinical and histopathological characteristics and response to treatment were assessed. RESULTS: Clinical features included meibomian gland dysfunction, blepharitis, chalazia, trichiasis, madarosis, conjunctivitis, chronic eyelid edema, and eyelid plaques. Histopathology showed hyperkeratotic epithelium, degeneration of the basal cell layer, and a perivascular lymphocytic infiltrate. There was delayed diagnosis in all cases, ranging from 4 months to 25 years. All of the patients responded to systemic hydroxychloroquine therapy. CONCLUSIONS: Heightened awareness of eyelid DLE may lead to earlier detection and specific therapy for this chronic disorder. PMID- 15878051 TI - How can we assure that neuro-ophthalmology will survive? PMID- 15878052 TI - Management of the patient with suspected temporal arteritis a decision-analytic approach. AB - OBJECTIVE: To perform a decision analysis of temporal arteritis (TA) to guide clinicians in the interpretation of diagnostic testing and choice of therapy. DESIGN: Computer-based decision analytic model. METHODS: A 785-node decision tree was created that reflects common testing and therapeutic options for a patient with suspected TA. A comprehensive literature search was then performed. From this search, point estimates and distributions for pooled probabilities and utilities were derived using inverse variance weighting and random effects techniques. Employing utility analysis, this decision model selects the diagnostic/therapeutic pathway resulting in the greatest utility for any user defined set of patient characteristics on presentation. MAIN OUTCOME MEASURE: Using utility analysis, the diagnostic/therapeutic pathway that results in the least expected disutility is selected as the optimal course of action. RESULTS: The choice of diagnostic testing depends on several factors, including patient age, symptoms, and clinical findings. These factors can be used to calculate the pretest probability of TA being present. The optimal selection of diagnostic tests (laboratory or biopsy) depends on the pretest probability of disease. A temporal artery biopsy is recommended under most circumstances, with the choice of a unilateral versus bilateral biopsy depending upon blood test results and calculated pretest probability. A few scenarios exist in which blood tests alone can rule in or rule out TA without the need for biopsy. Empiric steroid therapy is almost never recommended. CONCLUSIONS: In TA, both the disease and its treatment are hazardous for the patient. Clinicians should have a very low threshold to initiate a diagnostic workup for TA. Physicians are often uncertain when a temporal artery biopsy is indicated and whether to perform a unilateral or bilateral biopsy. Often, the pathway chosen is not evidence based. Although the biopsy has long been considered the gold standard for diagnosis, it is invasive and less than 100% sensitive. The decision whether to undertake unilateral or bilateral biopsies is difficult, and our decision model delineates a method for choosing. PMID- 15878053 TI - The diagnostic yield of the evaluation for isolated unexplained optic atrophy. AB - PURPOSE: To report the diagnostic yield for the evaluation of isolated and unexplained optic atrophy. SETTING: Two tertiary care academic neuroophthalmology clinics. DESIGN: Retrospective case series. PARTICIPANTS: Patients with optic atrophy. METHODS: Retrospective review of all charts with the diagnosis of optic atrophy. Included patients were adults with isolated, but unexplained, optic atrophy. Patients were excluded if they were children, had incomplete or inadequate documentation of the findings, had nonneurologically isolated optic atrophy (e.g., other localizing findings), or had a history (e.g., prior neuroimaging study showed a compressive lesion, prior ischemic optic neuropathy) or examination (e.g., central retinal artery occlusion) evidence for an etiology for the optic atrophy. MAIN OUTCOME MEASURE: Results of diagnostic evaluation. RESULTS: A total of 1110 charts with the diagnosis of optic atrophy were reviewed from the 2 participating institutions (368 from the University of Cincinnati and 742 from the University of Iowa). Of these 1110 charts, 91 (8%) with isolated unexplained optic atrophy were included, and 1019 charts (92%) were excluded. Of 91 included patients, 18 (20%) had a compressive lesion causing optic atrophy, and 73 (80%) cases had no etiology for the optic atrophy on neuroimaging. Of the 18 patients with abnormal imaging (e.g., meningioma, pituitary adenoma, craniopharyngioma) studies, 11 had bilateral and 7 had unilateral optic atrophy. Five of the 18 patients had progressive visual loss, 3 had hemianopic visual field loss, and 11 were younger than 50 years old. CONCLUSIONS: Patients with optic atrophy in our study typically had historical or examination findings that led to an etiologic diagnosis. Neuroimaging showed an etiology in 20% of patients. Other laboratory testing did not produce an etiologic diagnosis in the absence of a suggestive history or examination. On the basis of our results, we recommend neuroimaging for all patients with unexplained optic atrophy and consideration for directed laboratory studies only. PMID- 15878054 TI - Diabetes and hypertension in isolated sixth nerve palsy: a population-based study. AB - PURPOSE: Diabetes mellitus and systemic hypertension are frequently reported as ischemic causes of sixth nerve palsy/paresis, but there are few rigorous studies to support these associations. We conducted a population-based case-control study to determine the presence and magnitude of any association of preexisting diabetes mellitus and systemic hypertension with isolated sixth nerve palsy. DESIGN: Retrospective population-based case-control study. PARTICIPANTS AND CONTROLS: Participants were patients with new onset of neurologically isolated sixth nerve palsy or paresis (n = 76) in Olmsted County, Minnesota, from January 1, 1978, to December 31, 1992. Controls (n = 76) were selected from the same general population and were matched for age, gender, and length of medical follow up. METHODS: Using the Rochester Epidemiology Project medical records linkage system, which captures virtually all medical care provided to residents of Olmsted County, Minnesota, we identified all incident cases of neurologically isolated sixth nerve palsy/paresis (n = 76) among county residents between the given dates. An equal number (n = 76) of controls were randomly selected from the general population. We reviewed the entire medical record of each case and control, using stringent predetermined criteria to define the presence of diabetes mellitus and systemic hypertension. We compared the prevalence of diabetes and systemic hypertension between cases and controls by use of chi square tests, and we calculated odds ratios (OR) with 95% confidence intervals (CI). MAIN OUTCOME MEASURES: Presence or absence of diabetes mellitus and systemic hypertension. RESULTS: Diabetes mellitus occurred more frequently in cases (23.7%) than in controls (5.3%; P = 0.001; OR, 5.59; 95% CI, 1.79-17.42). Systemic hypertension occurred with similar frequency in cases (51.3%) and controls (39.5%; P = 0.14; OR, 1.62; 95% CI, 0.85-3.08). Coexistent diabetes mellitus and hypertension were more common in cases (18.4%) than in controls (2.6%; P = 0.002; OR, 8.36; 95% CI, 1.83-38.18). CONCLUSIONS: We conclude that there is a 6-fold increase in odds of having diabetes in cases of sixth nerve palsy over controls, whereas systemic hypertension does not seem to be associated with increased odds. In contrast, there is an 8-fold increased odds of having coexistent diabetes and hypertension in cases of sixth nerve palsy over controls. The much-cited association of systemic hypertension alone with sixth nerve palsy may be coincidental. PMID- 15878055 TI - Initial evaluation of subcutaneous daclizumab treatments for noninfectious uveitis: a multicenter noncomparative interventional case series. AB - PURPOSE: To assess the feasibility of a study design that may determine whether subcutaneous administration of the interleukin-2 receptor antibody daclizumab can safely reduce the dependence on standard systemic corticosteroids or other immunosuppressive regimens in patients with sight-threatening, noninfectious intermediate uveitis, posterior uveitis, or panuveitis. DESIGN: Prospective, multicenter, nonrandomized, noncomparative, open-label interventional trial. PARTICIPANTS: Fifteen patients, 5 each at 3 clinical centers, with noninfectious intermediate, posterior, or panuveitis, who require a currently stable immunosuppression regimen of systemic corticosteroids and/or other systemic treatments to control noninfectious intraocular inflammation. METHODS: After enrollment and baseline ophthalmic evaluations, 2 induction treatments were given 2 weeks apart using subcutaneous (SC) daclizumab at 2 mg/kg. Subcutaneous daclizumab maintenance treatments were then continued every 2 weeks at 1 mg/kg for 6 months. The initial immunosuppression load was tapered over 8 to 12 weeks in a staggered fashion beginning with the first induction treatment. Safety evaluations were performed at each treatment visit, with a primary efficacy evaluation at 12 weeks and a repeat efficacy evaluation at 26 weeks. MAIN OUTCOME MEASURES: Best-corrected visual acuity (Early Treatment of Diabetic Retinopathy Study [ETDRS] method) with a concurrent taper of concomitant systemic immunosuppression medication load (tabulated by use of a weighted scoring system) was assessed; target for success was defined as a 50% or greater reduction in concomitant immunosuppression load by 12 weeks while maintaining visual acuity within 5 ETDRS letters of baseline. Ocular inflammation was assessed at each visit with standardized grading scales. RESULTS: Ten of 15 patients (67%) receiving SC daclizumab treatments every other week successfully achieved the primary efficacy end point of reducing their concomitant immunosuppression load by at least 50% while maintaining their baseline visual acuity at 12 and 26 weeks. Subcutaneous daclizumab injections were well tolerated with no serious adverse events observed during the 6 months of treatments, although 3 patients experienced possibly related, nonserious adverse events. CONCLUSIONS: Subcutaneous daclizumab induction treatments at 2 mg/kg followed by 1 mg/kg maintenance treatments every other week seems safe and, in most cases, may reduce the concomitant immunosuppressive load required to treat noninfectious uveitis for 12 to 26 weeks. PMID- 15878056 TI - Risk factors for mortality in patients with AIDS in the era of highly active antiretroviral therapy. AB - OBJECTIVE: To evaluate risk factors for mortality among patients with AIDS in the era of highly active antiretroviral therapy (HAART), particularly the effect of cytomegalovirus (CMV). DESIGN: Prospective cohort study of patients with AIDS, conducted from 1998 through 2003. PARTICIPANTS: One thousand five hundred eighty three patients with AIDS, of whom 374 had CMV retinitis. METHODS: Patients were contacted every 3 months, with examinations at least every 6 months, in which standardized data were collected on AIDS history and treatment, eye examinations, and hematologic, virologic, and immunologic laboratory data. MAIN OUTCOME MEASURE: Mortality. RESULTS: The overall mortality rate was 0.07 deaths/person year. In a multivariate analysis, the following baseline risk factors were associated with an increased mortality: higher human immunodeficiency virus (HIV) viral load (relative risk [RR] = 4.6 for HIV viral load >100,000 copies/ml vs. <400 copies/ml; P<0.0001), lower CD4+ T-cell count at enrollment (RR = 3.8 for CD4+ T cell count 0-49 cells/microl vs. > or = 200 cells/microl; P<0.0001), CMV viral load > or = 400 copies/ml (RR = 1.9; P = 0.002), lower hemoglobin (RR = 1.7 for hemoglobin <10 g/dl; P = 0.009), a history of cryptococcal meningitis (RR = 1.7; P = 0.02), CMV retinitis (RR = 1.6; P = 0.0002), and Karnofsky score < or = 80 (RR = 1.4; P = 0.008). CONCLUSIONS: In the era of HAART, CMV disease as manifested by CMV retinitis and a detectable CMV viral load were associated with an increased risk for mortality, even after adjusting for demographic, treatment, immunologic, and HIV virologic factors. PMID- 15878057 TI - Pathological findings in eyes with the ganciclovir implant. AB - OBJECTIVE: To examine the pathologic findings of eyes treated with ganciclovir implants. DESIGN: Retrospective consecutive case series. PARTICIPANTS: Thirty three eyes from 19 AIDS patients with ganciclovir implants. METHODS: Pathologic specimens collected from 1993 through 1999 at the National Eye Institute were reviewed. Thirty-three eyes from 19 patients were identified with ganciclovir implants. The submitted eyes then were sectioned through the implant site and stained with hematoxylin and eosin, periodic acid-Schiff, and other special stains when needed. Medical records, including operative and postoperative notes, were reviewed carefully. MAIN OUTCOME MEASURE: Light microscopic findings at and around the site of implantation. RESULTS: Scars of surgical perforation were present in all eyes. Fibrous ingrowth developed from the implant site into the vitreous in 32 of the 33 eyes. Vitreous hemorrhage was present in 18 of the 33 eyes. Poor wound apposition was found in 2 of the 33 eyes, both of which had undergone multiple procedures. Foreign body giant cell reactions were observed in most of the eyes related to suture material. Thirty-two of the 37 implant sites were located within the pars plana, whereas the other 5 were either on the border of the pars plana and pars plicata (n = 4) or within the pars plicata (n = 1). Hyalinization, atrophic changes of the ciliary body in the area of implantation, or both were observed in 18 eyes. CONCLUSIONS: The ganciclovir implant is well tolerated within the eye. Fibrous ingrowth is present in most eyes and seems to be a benign occurrence because of its limited extension. Microscopic vitreous hemorrhage is present in many eyes, especially those that underwent multiple procedures. Poor wound apposition occurred rarely and was found only in eyes that had undergone multiple procedures. PMID- 15878058 TI - 14-year incidence, progression, and visual morbidity of age-related maculopathy: the Copenhagen City Eye Study. AB - PURPOSE: To describe the 14-year incidence of age-related maculopathy (ARM) lesions and the related visual loss. DESIGN: Population-based cohort study. PARTICIPANTS: Nine hundred forty-six residents (age range, 60-80 years) of Copenhagen participated in the study from 1986 through 1988. Excluding participants who had died since baseline, 359 persons (97.3% of survivors) were reexamined from 2000 through 2002. METHODS: Participants underwent extensive ophthalmologic examinations. Age-related maculopathy lesions were determined by grading color fundus photographs from the examinations using a modified Wisconsin Age-Related Maculopathy Grading System. MAIN OUTCOME MEASURES: Incidence of drusen type and size, pigmentary abnormalities, pure geographic atrophy, exudative ARM, visual impairment, and blindness. RESULTS: The 14-year incidences of early and late ARM were 31.5% and 14.8%, respectively. Individuals 75 to 80 years of age at baseline had significantly (P< or =0.05) higher 14-year incidences of the following lesions than those aged 60 to 64 years: medium or large drusen (> or =125 microm; 34.2% vs. 12.8%, respectively), soft drusen (45.2% vs. 21.4%), pigmentary abnormalities (31.4% vs. 17.0%), pure geographic atrophy (17.4% vs. 1.0%), and exudative ARM (23.3% vs. 5.7%). Severe drusen type, large drusen, and retinal pigmentary abnormalities at baseline were important predictors of incident late ARM. The 14-year incidences of visual impairment (<20/40 but >20/200) or legal blindness from late ARM were 6.0% and 3.4%, respectively. Late ARM caused 35.7% of all visual impairment and 66.7% of all blindness. CONCLUSIONS: There is a high incidence of ARM lesions in this elderly white population. Severe drusen type and size or a combination of drusen and pigmentary abnormalities significantly increases the risk of developing late ARM, the most frequent cause of legal blindness in this population. PMID- 15878059 TI - Hyperglycemia, blood pressure, and the 9-year incidence of diabetic retinopathy: the Barbados Eye Studies. AB - OBJECTIVES: To evaluate factors related to the incidence of diabetic retinopathy (DR) in a population of African descent, after 9 years of follow-up. DESIGN: Population-based cohort study; 81% participation after 9 years. PARTICIPANTS: Three hundred twenty-four participants of the Barbados Eye Studies, with diabetes mellitus (DM) at baseline and at risk for developing DR during follow-up. METHODS: Diabetes-related changes were assessed by masked gradings of baseline and follow-up photographs using a standardized system. The 9-year cumulative incidence of DR was based on participants with DM and free of retinopathy at baseline; incidence rates were estimated by the product-limit approach. Cox regression models for discrete-time data were used to evaluate risk factors associated with the 9-year incidence of DR. RESULTS: Multivariate analyses revealed that older age at DM onset decreased the 9-year risk of DR development; for each 10 years of older age at onset, the risk of DR decreased by 30% (risk ratio [RR], 0.7; 95% confidence interval [CI], 0.56-0.96). The risk of DR doubled among persons with DM duration between 5 and 9 years (RR, 2.1; 95% CI, 1.2-3.6) versus those with shorter durations; it also doubled in those treated with oral medications or insulin at baseline versus those treated with diet only. Antihypertensive treatment halved the risk of DR versus no treatment (RR, 0.5; 95% CI, 0.3-0.9) and high systolic or diastolic blood pressure (BP) increased risk. Thus, DR risk increased by 30% for every 10 mmHg of higher systolic BP at baseline (RR, 1.3; 95% CI, 1.1-1.4) or of BP increase from baseline to the 4-year follow-up (RR, 1.3; 95% CI, 1.1-1.4). Diabetic retinopathy risk similarly increased with each 1% of higher glycosylated hemoglobin level at baseline (RR, 1.3; 95% CI, 1.2-1.5). CONCLUSIONS: The long-term follow-up of persons with DM in this population of African origin, where disease prevalence is high, identified important potentially modifiable risk factors for DR. Findings suggest that efforts to achieve optimal glycemic and BP control may reduce the vision threatening complications of DM. PMID- 15878060 TI - Vitreous levels of vascular endothelial growth factor and intercellular adhesion molecule 1 are related to diabetic macular edema. AB - PURPOSE: To investigate whether vascular endothelial growth factor (VEGF) or soluble intercellular adhesion molecule 1 (sICAM-1) is related to diabetic macular edema (DME). DESIGN: Retrospective case-control study. PARTICIPANTS: Thirty-three patients who had DME and 13 patients with nondiabetic ocular disease (control group). METHODS: Vitreous fluid samples were obtained at the time of vitreoretinal surgery, and the VEGF and sICAM-1 levels in vitreous fluid and plasma were measured by enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Relationship between vitreous levels of VEGF or sICAM-1 and DME. RESULTS: Vitreous levels of VEGF and sICAM-1 were significantly higher in DME patients than in control patients (P < 0.0001 and P = 0.0004, respectively). The vitreous level of VEGF was significantly correlated with that of sICAM-1 (P < 0.0001). Vitreous levels of both VEGF and sICAM-1 were significantly higher in patients with hyperfluorescent DME than in those with minimally fluorescent DME (P = 0.0027 and P = 0.0005, respectively). The vitreous levels of both VEGF and sICAM 1 were significantly correlated with retinal thickness at the central fovea (P < 0.0001 and P = 0.0005, respectively) CONCLUSIONS: We found that both VEGF and sICAM-1 were elevated in the vitreous fluid of patients with hyperfluorescent DME. These results suggest that VEGF and ICAM-1 are related to the increase of vascular permeability in DME patients. PMID- 15878061 TI - Outcomes of 140 consecutive cases of 25-gauge transconjunctival surgery for posterior segment disease. AB - PURPOSE: To evaluate the safety and efficacy of 25-gauge instrumentation for a variety of vitreoretinal conditions on previously nonvitrectomized eyes. DESIGN: Single-center, retrospective, interventional case series. PARTICIPANTS: One hundred forty eyes of 140 patients were evaluated at the Doheny Retina Institute from July 2002 to July 2003. INTERVENTION: All patients underwent surgical procedures using the Millennium 25-gauge Transconjunctival Standard Vitrectomy system. Twenty eyes (14.3%) underwent procedures without vitrectomy. MAIN OUTCOME MEASURES: Postoperative visual acuity (VA), intraocular pressure, surgical time, postoperative inflammation, complications, and number of sutured sites. RESULTS: No intraoperative complications were noted. No cases required conversion to 20 gauge machines. Ten cases (7.1%) involved single-site sclerotomy suture placement due to bleb formation at the conclusion of the procedure, but 5 of these entry sites were enlarged to facilitate larger instrumentation for tissue manipulation. Median VA improved from 20/250 (logarithm of the minimum angle of resolution, 1.08+/-0.47) preoperatively to 20/60 (0.47+/-0.30) (P<0.0001) at final visit. Mean follow-up was 33.8+/-9.7 weeks, and all eyes were observed for a minimum of 12 weeks. Mean total surgical time was 17.4+/-6.9 minutes. Intraocular pressures remained stable throughout the postoperative course. Five eyes (3.8%) presented on day 1 with shallow choroidal detachments, but all resolved by day 7, and none required volume infusion during the postoperative period. All but one of these cases was within the first 50 procedures performed. No detectable inflammation was noted in any eyes by 4 weeks postoperatively. No case of retinal detachment or endophthalmitis was recorded. CONCLUSIONS: Transconjunctival surgery using 25 gauge instrumentation may hasten postoperative recovery by decreasing overall surgical time and postoperative inflammation. Procedures requiring minimal intraocular manipulation did not require sutures and, thus, may be better suited for this surgical modality. PMID- 15878062 TI - Fundus autofluorescence and central serous chorioretinopathy. AB - PURPOSE: To investigate the autofluorescence characteristics in patients with central serous chorioretinopathy. DESIGN: Observational case series. PARTICIPANTS: Thirty consecutive patients examined in a private referral practice. METHODS: Patients were imaged with autofluorescence photography, fundus photography, fluorescein angiography, and optical coherence tomography (OCT). The mean and standard deviation (SD) of the grayscale values from a 100-pixel diameter circle centered on the fovea were obtained and normalized with the level of autofluorescence of the posterior pole. RESULTS: There were 30 patients, 23 male (76.7%) and 7 female (23.3%), with a median visual acuity (VA) of 20/25 and a range of 20/15 to 20/400. Stepwise linear regression that included individual fixed effects found that normalized central macular autofluorescence (P < 0.001), pigment mottling in the fovea (P = 0.045), subfoveal fluid detected by OCT (P = 0.033), and the SD of the central macular autofluorescence (P = 0.025) produced a highly significant model (R2 = 0.92, P < < 0.001) predicting VA. Increasing levels of autofluorescence were correlated with accumulation of material on the outer surface of the retina as seen by OCT. Decreased central macular autofluorescence, particularly in those eyes with central geographic retinal pigment epithelial atrophy, was associated with poor VA. CONCLUSIONS: This study established that autofluorescence changes occurring in central serous chorioretinopathy with explicit patterns can be measured in a noninvasive manner, and this information can be used to estimate the damage induced by central serous chorioretinopathy with a high degree of statistical significance. We hypothesize that the material on the outer surface of the elevated retina may represent accumulation of photoreceptor outer segments secondary to the lack of direct apposition and phagocytosis by the retinal pigment epithelium. PMID- 15878063 TI - Ruthenium brachytherapy for uveal melanoma, 1979-2003: survival and functional outcomes in the Swedish population. AB - PURPOSE: To evaluate observed and relative survival rates, enucleation rates, and visual outcome after ruthenium 106 brachytherapy for uveal melanoma. DESIGN: Retrospective cases series from the Swedish national referral center. PARTICIPANTS: Five hundred seventy-nine patients (579 eyes) with choroidal or ciliary body melanomas, including 55 tumors more than 7 mm in height, treated with ruthenium episcleral plaques from January, 1979, through April, 2003. METHODS: Clinical and radiotherapy data were extracted from a dedicated database, and survival status was determined through population registries. Tumor size was classified according to the Collaborative Ocular Melanoma Study criteria. The 5- and 10-year relative survival rates were estimated, and univariate and multivariate regression models were constructed for predictive factors on observed survival, enucleation, and visual deterioration. MAIN OUTCOME MEASURES: Observed and relative survival rate, proportion of secondary enucleation, deterioration of visual acuity to less than 0.5, respectively, to 0.1 or worse. RESULTS: Tumors were classified as small in 10.5%, medium in 78.4%, and large in 9.2% of patients. The 5- and 10-year observed overall survival rates were 83.3% and 71.5%, respectively, and the corresponding relative rates were 95.5% and 94%, respectively. Factors predicting survival were tumor diameter, patient age, and secondary enucleation. One hundred six patients (18%) underwent enucleation up to 14 years after plaque treatment. The only predictive factor for enucleation was tumor size. At 5 years, 31% of the patients retained 0.5 visual acuity or better, and 49% retained better than 0.1 visual acuity. Predictive factors for visual deterioration were visual acuity and distance from posterior tumor border to the foveola. CONCLUSIONS: After ruthenium brachytherapy for uveal melanoma, the survival rates and visual outcomes in this population-based investigation were similar to previously published results. The eye was retained in 81.7% of patients. Careful patient selection (presently we only treat melanomas 7 mm or smaller in height) and life-long monitoring for recurrences is warranted. PMID- 15878064 TI - Grouped congenital hypertrophy of the retinal pigment epithelium follows developmental patterns of pigmentary mosaicism. AB - PURPOSE: To determine whether sectorial-oriented grouped pigmentations of the retina follow developmental patterns of pigmentary mosaicism. DESIGN: Systematic literature review. PARTICIPANTS: Fundus images from patients with grouped congenital hypertrophy of the retinal pigment epithelium (CHRPE). METHODS: An extensive Internet and library search was performed to obtain articles dealing with grouped CHRPE. Each article was carefully screened for fundus images and inclusion criteria. Fundus images of sufficient quality were scanned, digitized, and matched in size using Adobe Photo Shop to compare the pattern location and extension of the pigmented lesion. These obtained patterns of grouped CHRPE were than mapped and superimposed. RESULTS: Forty-five images were retrieved from 32 articles with grouped CHRPE. The lesions extended from the margin of the optic disc and radiated in sectors to the fundus periphery. The stream of growth did not follow the pattern of the retinal nerve fiber layer, because the clusters of pigmented cells crossed the midline raphe not sparing the macular area. Smaller lesion clusters were mainly located near the optic disc, whereas larger lesions were found in the periphery. CONCLUSIONS: The growth pattern of grouped CHRPE is similar to cutaneous sectorial pigmentations. We speculate that pigmentary mosaicisms may be a modified wild-type allele in a somatic cell clone during early embryogenesis following developmental lines analogous to the cutaneous lines of Blaschko. The sectorial pigmentations on the ocular fundus may reflect the stream, outgrowth, and migration of retinal pigment epithelium cells during embryogenesis. PMID- 15878065 TI - Fundus perimetry with the Micro Perimeter 1 in normal individuals: comparison with conventional threshold perimetry. AB - PURPOSE: To determine differential light threshold values obtained with the Micro Perimeter 1 (MP1) in healthy volunteers and to correlate them with conventional automated static threshold perimetry using the Octopus 101 Perimeter. DESIGN: Prospective comparative observational study. PARTICIPANTS: Thirty healthy volunteers. METHODS: In 30 eyes of 30 healthy volunteers, static threshold perimetry was performed with the MP1 Micro Perimeter (Nidek Inc., Italy) and the Octopus 101 (Haag-Streit AG, Switzerland) in the same eye in random order. MAIN OUTCOME MEASURES: Differential light threshold values obtained with the MP1 and their difference to differential light threshold values with the Octopus. Differential light sensitivity was compared for 21 matching points in a rectangular test grid using similar examination settings with Goldmann III stimuli, stimulus presentation time of 100 msec, and white background illumination (1.27 cd/m2). RESULTS: For the 21 matching locations, mean differential light thresholds with the MP 1 and the Octopus were 15.5+/-0.8 decibels (dB) (range, 13.0-17.1) and 30.2+/-1.2 dB (range, 27.7-32.0), respectively. On the average, the Octopus showed higher threshold values for all test locations than the MP1. The mean difference between both examinations was 14.6+/-1.8 dB for all locations and 14.8+/-1.7 dB excluding the test locations at the blind spot. With a considerably high grade of variation according to the test point location, the difference between the 2 devices varied from 11.4 to 18.3 dB, showing a vertical asymmetry with a larger difference in the lower part of the visual field. Linear regression of the perimetric results for each test point location, excluding the area of the blind spot and the lower line of the test grid, showed significant correlation (r = 0.56; P = 0.036). CONCLUSIONS: The results show that the MP1 provides reproducible threshold values with a systematic difference compared with standard Octopus perimetry of approximately 15 dB. With a larger difference in the lower part of the visual field, differential light sensitivity values in microperimetry with the MP1 are comparable to the threshold values obtained with the Octopus 101 using a correction factor of 11.4 to 18.3 dB according to stimulus location. PMID- 15878066 TI - Discrimination of glaucomatous optic neuropathy by digital stereoscopic analysis. AB - PURPOSE: To evaluate the diagnostic power of a novel digital stereoscopic imaging system in the diagnosis of glaucomatous optic neuropathy. DESIGN: Prospective cross-sectional analysis of the diagnostic accuracy of digital stereoscopic optic disc analysis in the diagnosis of glaucomatous optic neuropathy exhibiting mild to moderate field loss. PARTICIPANTS: Fifty-two patients with open-angle glaucoma and 54 normal individuals were recruited. The presence of a reproducible visual field loss characteristic of glaucoma was used as the reference standard for the presence of glaucoma independent of the optic nerve head appearance. Patients were excluded if the optic disc, fundus, or visual field indicated other disease. One eye from each patient and individual was included in the study, the eye with the least field loss and a randomly designated normal eye, respectively. METHODS: Simultaneous stereoscopic optic disc photography was performed on each specified eye. Three experienced observers viewed the resultant stereoscopic image of each nerve head using a Z screen, recorded a subjective clinical diagnosis, and undertook digital stereoscopic planimetry. Separate linear regression analysis was performed, post hoc, from the planimetric results for each observer of the logarithm of neuroretinal rim (NRR) against optic disc area derived from each normal eye. Eyes with NRR areas below the 95th prediction interval of the normal cohort were then classified as glaucomatous. MAIN OUTCOME MEASURES: Sensitivity and specificity for the detection of glaucomatous optic neuropathy. RESULTS: With subjective stereoscopic analysis, sensitivity for glaucoma detection among the 3 observers was 80.8%, 76.9%, and 90.4%, with respective specificities of 94.4%, 79.6%, and 79.6%. Regression analysis of the NRR in 30 degrees segments gave sensitivities between 69.2% and 80.8% and specificities between 83.3% and 90.7%. A combination of the subjective and quantitative analysis did not significantly improve discrimination. CONCLUSIONS: The subjective analysis of digital stereoscopic images provides a useful method for the discrimination of normal and glaucomatous optic nerves. Planimetric analysis does not significantly improve the diagnostic precision of this technique. PMID- 15878067 TI - Does adjunctive glaucoma therapy affect adherence to the initial primary therapy? AB - OBJECTIVE: To examine the effect of adding complexity to a glaucoma medical treatment regimen--specifically, what would occur to the refill rate (and, by inference, to adherence) when a second medication was added to a currently used once-daily drug. DESIGN: Open-label retrospective review of patient records. PARTICIPANTS: Patients of a large national health care provider who had received a prescription for latanoprost between July 1, 2001 and June 30, 2002. There were 1784 patients who had a second medication added and 3146 patients who remained on monotherapy. METHODS: For each patient, the mean number of days between refills was calculated for both the period before and that subsequent to the addition of the second medication, and an interperiod difference in refill interval between the 2 periods was calculated. Probability comparisons were performed using paired t tests (continuous) and Wilcoxon signed rank tests (categorical). RESULTS: The mean age of the population using second-line therapy was 68.3+/-14.5 years (range, 4-97), and 56% were female. In this population of 1784 patients who used 2 different ocular-hypotensive medications, mean refill intervals for latanoprost were 40.6+/-21.8 days before the addition of a second drug and 47.4+/-24.4 days after the addition of a second drug, with a mean increase of 6.7+/-25.6 days. For 22.9% (409/1784) of patients, the interval was increased by >2 weeks (P<0.0001). The mean refill interval was longer than that for the 3146 patients who continued on latanoprost monotherapy, which was 41+/-24 days. CONCLUSIONS: This statistically and clinically significant increase in refill intervals may affect intraocular pressure control. We suggest that, when adding a second drug, physicians need to consider the possible impact on the patient's adherence to the first drug. PMID- 15878068 TI - Safety and efficacy of phacoemulsification compared with manual small-incision cataract surgery by a randomized controlled clinical trial: six-week results. AB - OBJECTIVE: To compare the efficacy, safety, and refractive errors of astigmatism after cataract surgery by phacoemulsification and manual small-incision cataract surgery techniques. DESIGN: Masked randomized control clinical trial. PARTICIPANTS: Four hundred eyes of 400 patients, 1:1 randomization with half in each arm of the trial. METHODS: A total of 400 eyes was assigned randomly to either phacoemulsification or small-incision groups after informed consent and were operated on by 4 surgeons. They were masked to the technique of surgery before, during, and after cataract surgery and followed up to 1 year after surgery. The intraoperative and postoperative complications, uncorrected and best corrected visual acuity, and astigmatism were recorded at 1 and 6 weeks postoperatively. MAIN OUTCOME MEASURES: The proportion of patients achieving visual acuity better than or equal to 6/18 with and without spectacles after cataract surgery in the operated eye up to 6 weeks, postoperative astigmatism, and complications during and after surgery. RESULTS: This article reports clinical outcomes up to 6 weeks. Three hundred eighty-three of 400 (95.75%) patients completed the 1-week follow-up, and 372 of 400 (93%) patients completed the 6-week follow-up. One hundred thirty-one of 192 (68.2%) patients in the phacoemulsification group and 117 of 191 (61.25%) patients in the small-incision group had uncorrected visual acuity better than or equal to 6/18 at 1 week (P = 0.153). One hundred fifty of 185 (81.08%) patients of the phacoemulsification group and 133 of 187 (71.1%) patients of the small-incision group (P = 0.038) were better than or equal to 6/18 at the 6-week follow-up for presenting visual activity. Visual acuity improved to > or = 6/18 with best correction in 182 of 185 patients (98.4%) and 184 of 187 (98.4%) patients (P = 0.549), respectively. Poor outcome (postoperative visual acuity < 6/60) was noted in 1 of 185 (0.5%) in the phacoemulsification group and none in the small-incision group. The mode of astigmatism was 0.5 diopters (D) for the phacoemulsification group and 1.5 D for the small-incision group, and the average astigmatism was 1.1 D and 1.2 D, respectively. There was an intra-surgeon variation in astigmatism. The phacoemulsification group had 7 posterior capsular rents compared with 12 in the small-incision group, but the phacoemulsification group had more corneal edema on the first postoperative day. CONCLUSIONS: Both the phacoemulsification and the small-incision techniques are safe and effective for visual rehabilitation of cataract patients, although phacoemulsification gives better uncorrected visual acuity in a larger proportion of patients at 6 weeks. PMID- 15878069 TI - Anterior vitreous tapping to manage positive vitreous pressure during triple procedures. AB - PURPOSE: To describe the technique and outcome of pars plana vitreous aspiration during penetrating keratoplasty (PK) combined with cataract extraction (CE) and intraocular lens (IOL) implantation (triple procedure) to prevent positive vitreous pressure and its associated complications. DESIGN: Nonrandomized prospective noncomparative interventional case series. PARTICIPANTS: Sixty-five patients who underwent triple procedures between June 2001 and September 2003. METHODS: Triple procedures performed on all 65 patients consisted of PK, extracapsular CE, and IOL implantation with standard techniques. After nonpenetrating trephination, anterior vitreous tapping was performed with a 23 gauge needle on a 5-ml syringe over the pars plana 3.5 mm posterior to the limbus in the superotemporal quadrant. MAIN OUTCOME MEASURES: Preoperative best corrected visual acuity (BCVA), volume of aspirated liquefied vitreous, intraoperative and postoperative complications, and postoperative BCVA at 3 and 6 months. RESULTS: Corneal scarring with cataract was the most common indication for surgery. Preoperative BCVA ranged from 20/160 to light perception. Aspirated vitreous volume varied from 0.3 to 1.5 ml, averaging 1.0 ml. No case had evidence of positive vitreous pressure during surgery. The mean follow-up period was 9.6+/ 2.6 months (range, 6-24). Postoperative complications consisted of graft rejection in 4 cases (6.2%), primary graft failure in 3 (4.6%), and secondary glaucoma in 1 (1.5%). Of the patients, 70.8% had a postoperative BCVA of > or =20/160 at 6 months. Vitreous hemorrhage, retinal tearing, and detachment were not observed. CONCLUSIONS: Pars plana anterior vitreous tapping is a safe adjunct for triple procedures to prevent positive vitreous pressure and its associated complications. PMID- 15878070 TI - Role of hyaluronidase in diplopia after peribulbar anesthesia for cataract surgery. AB - OBJECTIVE: To determine the protective action of hyaluronidase on peribulbar anesthesia-related diplopia in patients undergoing cataract surgery. DESIGN: Single-center observational case series. PARTICIPANTS: All patients undergoing elective phacoemulsification and intraocular lens implantation under peribulbar anesthesia between February 2001 and January 2003. METHODS: We compared the incidence of postoperative diplopia between 2 periods--February 2001 to January 2002 (P1) and February 2002 to January 2003 (P2)--which differed by the presence (P1) or absence (P2) of hyaluronidase in the anesthetic solution. MAIN OUTCOME MEASURES: All patients were examined on the first and fifth postoperative days during both periods. When diplopia was diagnosed, we recorded the characteristics of the patient, peribulbar anesthesia, and diplopia (orthoptic examination, and magnetic resonance imaging in some cases). RESULTS: Seven thousand two hundred five patients were studied. During P1, 3582 patients received peribulbar anesthesia, and no cases of diplopia occurred. During P2, 3623 patients received peribulbar anesthesia, and 27 cases of diplopia occurred (incidence, 0.75%; P = 0.0002 vs. P1). Diplopia involved the inferior rectus (40%) and the external rectus (37%) muscles. Diplopia was persistent in 54% of the cases. CONCLUSIONS: Peribulbar anesthesia-related diplopia was significantly more frequent when hyaluronidase was not added to the anesthetic solution. PMID- 15878071 TI - Detection of virulence factors in a corneal isolate of Klebsiella pneumoniae. AB - OBJECTIVE: To report on the microbiological findings of a Klebsiella pneumoniae strain isolated from a patient with keratitis. DESIGN: Interventional case report. INTERVENTION AND TESTING: Conjunctival swabs and corneal scrapings from the right eye were inoculated for culture. The isolate was analyzed for the presence of the mucoid phenotype and the ability to form biofilm. We also investigated whether the formation of biofilm by the corneal Klebsiella isolate is affected by N-acetylcysteine. MAIN OUTCOME MEASURES: Culture results and biofilm production were analyzed. RESULTS: K. pneumoniae was grown from the conjunctiva and cornea. The isolate showed the mucoid phenotype and strong biofilm production. N-acetylcysteine had an inhibitory effect on both biofilm formation and preformed biofilm. CONCLUSIONS: K. pneumoniae can cause severe keratitis. The presence of virulence factors, such as the mucoid phenotype and the ability to form biofilm, may be important in determining corneal infection. N acetylcysteine is a potential candidate for use as an inhibitor of Klebsiella biofilm formation. PMID- 15878072 TI - Incidence of herpes simplex virus keratitis in France. AB - OBJECTIVE: To assess the incidence and clinical characteristics of herpes simplex keratitis in France. DESIGN: National multicenter prospective study on herpetic keratitis. PARTICIPANTS: During a 3-month study period (September-December 2002), all cases of herpes simplex keratitis were prospectively reported by a randomly selected sample of 412 ophthalmologists representative of the 5471 French ophthalmologists in terms of gender, geographic distribution, and clinical practice. METHODS: The following set of assumptions were made to calculate the incidence of herpectic keratitis: (1) the participating (self-selected) ophthalmologists in the study could adequately represent French ophthalmologists, (2) estimates based on the 3-month study period could be used to calculate the annual incidence (i.e., no significant seasonal variations), and (3) all patients suffering from herpetic corneal lesions consult or are referred to an ophthalmologist. We calculated the incidence of herpetic keratitis by (1) estimating the average number of incident cases per ophthalmologist per year, (2) multiplying the average number of incident cases by the total number of French ophthalmologists (n = 5471), and (3) dividing the result by the French population. RESULTS: Four hundred twelve ophthalmologists reported 357 cases of herpes keratitis. We estimated that the overall incidence of herpetic keratitis during the study period was 31.5 per 100,000 person-years (95% confidence interval [CI], 25.5-37.5), and incidences were 13.2 per 100,000 person-years for new cases (95% CI, 10.4-15.9) and 18.3 per 100,000 person-years for recurrences (95% CI, 14.6-22.1). The most frequent types were dendritic keratitis (56.3%, n = 153), stromal keratitis (29.5%, n = 81), and geographic keratitis (9.8%, n = 27). Other ocular lesions were associated with keratitis in 35.0% (n = 125) of cases; the most frequent were conjunctivitis (18.8%, n = 67), uveitis (11.8%, n = 42), and/or lid involvement (8.6%, n = 31). CONCLUSIONS: This prospective epidemiological study provides an estimate of the incidence of herpes keratitis in France. Herpetic keratitis remains an epidemiologically important eye disease that justifies the need to pursue health care and research programs aimed at improving the outcome of ocular herpetic disease. PMID- 15878073 TI - Intraoperative mitomycin C and amniotic membrane transplantation for fornix reconstruction in severe cicatricial ocular surface diseases. AB - PURPOSE: To investigate whether intraoperative application of mitomycin C may enhance the success of amniotic membrane transplantation in symblepharon lysis and fornix reconstruction in severe cicatricial ocular surface diseases. DESIGN: Noncomparative interventional case series. PARTICIPANTS: Sixteen patients (8 female, 8 male; 18 eyes) with a mean age of 41+/-23.4 years (range, 3-79) and suffering from severe chemical/thermal burns (7 eyes), multiple recurrent pterygia and pseudopterygia (5 eyes), Stevens-Johnson syndrome (4 eyes), and ocular cicatricial pemphigoid (2 eyes) were consecutively enrolled. All except for 2 eyes had had prior surgical attempts of surgical reconstruction, including 6 eyes with a mucous membrane graft (MMG), but still presented with symblepharon and persistent ocular surface inflammation. INTERVENTION: After excision of subconjunctival fibrovascular tissues, 0.04% mitomycin C was applied for 5 minutes in the deep fornix before amniotic membrane transplantation. MAIN OUTCOME MEASURES: Deeper fornix, noninflamed ocular surface, and full motility. RESULTS: The mean epithelial healing time was 4.2+/-1.9 weeks. During the follow-up of 14.16+/-5.2 months, all eyes showed a marked reduction of conjunctival inflammation, a deep fornix, and a continuous tear meniscus. Of 12 eyes with motility restriction, 2 eyes with multiple recurrent pterygia and 1 eye with severe thermal burn showed recurrence of partial motility restriction 2 months after surgery. The vision of 9 eyes was successfully restored by an additional keratolimbal allograft with subsequent penetrating keratoplasty (6 eyes). CONCLUSION: Intraoperative application of mitomycin C is an effective means to reduce chronic and deep-seated conjunctival inflammation, and helps amniotic membrane restore a deep fornix after symblepharon lysis, even in eyes that had a failed MMG. Restoration of deep fornix and tear meniscus is an important prerequisite to achieve successful reconstruction by subsequent limbal stem cell transplantation. PMID- 15878074 TI - Correlation of corneal complications with eyelid cicatricial pathologies in patients with Stevens-Johnson syndrome and toxic epidermal necrolysis syndrome. AB - PURPOSE: To look at the correlation between many factors (time of hospitalization, floppy eyelid syndrome, trichiasis, open lacrimal puncta, symblepharon, and aqueous tear deficiency) and corneal complications in Stevens Johnson syndrome (SJS). DESIGN: Observational cases series. PATIENTS: Clinical data were retrospectively reviewed from 38 patients (32.7+/-20.1 years old) with SJS (n = 11) and with toxic epidermal necrolysis (TENS) (n = 27) from January 2002 to August 2004. One case report with SJS was included to verify the presence of tarsal/lid margin ulceration at the acute stage. METHODS: The medical history was retrieved regarding presumed causative medications used within 15 days and the duration of hospitalization. Data of the latest photographic documentation and eye examination were compared and correlated in a masked fashion. MAIN OUTCOME MEASURES: Floppy eyelid, trichiasis, lid margin keratinization, meibomian gland orifice metaplasia, symblepharon, tarsal scar, and corneal complications. RESULTS: Acute SJS/TENS was characterized by tarsal conjunctival ulceration. Keratinization of the eyelid margin with variable degrees of meibomian gland dysfunction was observed in all cases. Floppy eyelid, trichiasis, partially or totally opened lacrimal punctum, symblepharon, and aqueous tear deficiency were not significantly correlated with corneal complications. In contrast, there was a strong correlation between the severity of eyelid margin and tarsal pathology and the extent of corneal complications (Spearman r, 0.54; P = 0.0005). A multivariable regression analysis also showed that the extent of eyelid and tarsal pathology had a significant effect on corneal complications (coefficient, 0.84; P = 0.006). CONCLUSIONS: Patients with acute SJS/TENS are characterized by severe inflammation and ulceration of the tarsal conjunctiva and lid margins. If left unattended, lid margin keratinization and tarsal scar, together with lipid tear deficiency, contribute to corneal complications because of blink-related microtrauma. Attempts to suppress inflammation and scarring by amniotic membrane transplantation at the acute stage and to prevent microtrauma at the chronic stage are vital to avoid sight-threatening complications. PMID- 15878075 TI - Intralesional triamcinolone acetonide injection for primary and recurrent chalazia: is it really effective? AB - PURPOSE: To evaluate the safety and efficacy of intralesional triamcinolone acetonide (TA) injection in primary and recurrent chalazia. DESIGN: Retrospective, interventional, consecutive case series. PARTICIPANTS: One hundred forty-seven patients with primary or recurrent chalazia (155 cases) treated at the oculoplastic clinic at the Jules Stein Eye Institute between January 1, 2000, and December 31, 2003. METHODS: Patients received an intralesional injection of 0.1 to 0.2 ml TA (40 mg/ml). Data regarding lesion size, including digital color photography, lesion regression or recurrence, and complete ophthalmic examination, were recorded at the time of injection and at different intervals until resolution or surgical excision. Success was defined as at least an 80% decrease in size with no recurrence. If the lesion recurred or regression was minimal (<50%), further injections were given as needed. Patients who declined injection or who did not respond to 2 to 3 injections were referred for surgical excision and drainage. MAIN OUTCOME MEASURES: Lesion size, clinical resolution, time to resolution, recurrence, and complications. RESULTS: Most of the patients received 1 injection (93 patients; 60%) or 2 injections (31 patients; 20%) with resolution of the lesion (more than 80% decrease in size), with an average time to resolution of 2.5 weeks. Patients who did not respond to 2 injections were more likely to fail treatment (minimal or no regression), to respond to further injections, or to undergo surgical excision and drainage (P = 0.0001, chi-square test). Patients with blepharitis required more injections to resolution (2+/-1.3 vs. 1.4+/-1; P = 0.05, independent samples t test). Intraocular pressure and visual acuity remained stable after treatment. No complications, such as visual loss, subcutaneous fat atrophy, or skin depigmentation changes, were noted with steroids injections; assuming a complication rate of 2%, our power was adequate to rule out these complications. CONCLUSIONS: Intralesional TA injection in primary and recurrent chalazia is effective in achieving lesion regression. Most cases resolve with an average of 1 to 2 injections. Chalazia that fail to respond to 2 or 3 injections are more likely to benefit from surgical excision. It may be considered as a first treatment in cases where diagnosis is straightforward. PMID- 15878076 TI - Paradoxical use of frontalis muscle and the possible role of botulinum a toxin in permanent motor relearning. AB - PURPOSE: To report 7 patients with paradoxical use of the frontalis muscle despite postsurgical correction of ptosis with good postoperative eyelid position. Successful treatment with botulinum A toxin facilitated motor relearning and cessation of muscle contraction. DESIGN: Interventional case series. PARTICIPANTS: Seven patients, in 2 eye-plastic clinics, who underwent successful surgical correction of upper eyelid ptosis. METHODS: Review of clinical history, clinical photographs, treatment, and follow-up. MAIN OUTCOME MEASURES: Frontalis muscle contraction and upper eyelid position. RESULTS: Patients underwent successful surgical correction of ptosis but continued using the frontalis muscle despite good eyelid position postoperatively. Frontalis contraction ceased spontaneously in 2 patients, but required botulinum A toxin injection in 5. The effects of a single treatment of botulinum A toxin lasted from 3 months to 2 years, longer than the expected effect of the toxin. CONCLUSION: Patients with long-standing eyelid ptosis may paradoxically continue utilizing the frontalis after successful surgical correction and despite good postoperative eyelid position. Cessation of frontalis contraction can be achieved with a single injection of botulinum A toxin. We hypothesize that chemodenervation, achieved with the toxin, may influence the central nervous system to relearn the set point for muscle contraction and may be associated with permanent motor relearning. Spontaneous resolution of muscle contraction can occur in the first months after surgery. PMID- 15878077 TI - Simultaneous orbital decompression and correction of upper eyelid retraction versus staged procedures in thyroid-related orbitopathy. AB - PURPOSE: To evaluate the outcome of eyelid retraction surgery in thyroid-related orbitopathy (TRO) patients in 2 different surgical settings: done simultaneously with orbital decompression or as a staged procedure after orbital decompression. DESIGN: Retrospective, comparative, nonrandomized clinical study. PARTICIPANTS: Ninety-six patients (158 eyes). METHODS: A review of electronic medical records of TRO patients who underwent surgery for upper eyelid retraction and orbital decompression at the Jules Stein Eye Institute in 1999 to 2003 was performed. Data regarding eyelid position, comprehensive eye examination, surgical outcome, and complications were analyzed. MAIN OUTCOME MEASURES: Anatomical and functional success based on margin reflex distance (MRD1; < or = 5 mm was graded as mild retraction; > 5 mm and < 7 mm, moderate; and > 7 mm, severe), and patients' discomfort. RESULTS: One hundred fifty-eight eyelid retraction surgeries were performed on 96 TRO patients (18 male and 78 female; mean age, 48 years); mean follow up time was 15 (+/-12) months. Group 1 consisted of patients undergoing simultaneous eyelid retraction surgery and orbital decompression and comprised 97 cases (surgeries). Group 2 included 61 cases of staged surgery: orbital decompression and eyelid retraction at a later stage. The groups had similar surgical outcomes, and > 85% had a better eyelid position postoperatively. Reoperation rates for residual or recurrent eyelid retraction were similar, overcorrection was higher in group 2 (5% vs. 0%, P = 0.03). Changes in MRD1, lagophthalmos, and exophthalmos were similar (P > 0.05, independent samples t test). Correction of eyelid retraction was effective in treating patients' discomfort and exposure keratopathy (P = 0.04, chi2). No severe complications occurred after orbital decompression or eyelid retraction surgery in this group of patients. CONCLUSIONS: Transconjunctival Muller's muscle recession for correction of eyelid retraction in mild to moderate TRO patients, performed simultaneously with deep lateral wall orbital decompression, resulted in acceptable eyelid position in two thirds of our patients. Overcorrection and consecutive ptosis occurred less often after combined orbital decompression and eyelid retraction surgery than after isolated eyelid repositioning surgery. If confirmed in prospective controlled studies, eyelid-repositioning surgery performed at the time of orbital decompression may decrease the number of total procedures and compress the time needed for surgical rehabilitation. PMID- 15878078 TI - Combined rectus muscle transposition with posterior fixation sutures for the treatment of double-elevator palsy. AB - PURPOSE: To evaluate the surgical and functional results of full horizontal tendon rectus muscle transposition to the superior rectus (SR) insertion, augmented by posterior fixation sutures, in patients with double-elevator palsy (DEP). DESIGN: Retrospective nonconcurrent interventional comparative case series. PARTICIPANTS: Fourteen consecutive patients with DEP. INTERVENTION: Six patients treated for DEP by vertical transposition of the horizontal muscles to the SR insertion (Knapp procedure) were compared with 8 patients treated by the Knapp procedure combined with fixation of the transposed muscles to the sclera, adjacent to the SR, with nonabsorbable sutures. MAIN OUTCOME MEASURES: Postoperative ocular alignment, ductions, binocular functions, and rate of reoperation. RESULTS: Ocular deviation: Mean distance and near deviations decreased by 84% and 83%, respectively (P = 0.012), in the augmented-surgery group versus 48% and 47%, respectively (P = 0.03), in the standard-surgery group. Duction: Mean elevation deficiency in abduction and adduction improved by 64% and 65%, respectively (P = 0.01), in the augmented surgery group versus 42% and 55% (P = 0.02) in the standard group. Binocular functions: 3 patients (37%), all in the study group, gained binocular function. Reoperation was required in 5 patients (83.3%) in the control group. The difference in postoperative improvement between the groups was statistically significant for all 4 parameters. No postoperative complications or duction anomalies were observed during the follow-up period of 15.4 months (standard deviation, 5.5). CONCLUSIONS: The augmented Knapp procedure with superior posterior fixation suture is the preferred surgical treatment for patients with DEP. Its use in this series avoided the need for multiple surgeries on other extraocular muscles. PMID- 15878079 TI - Full vertical rectus muscle transposition combined with medial posterior fixation sutures for patients with adduction deficiency. AB - PURPOSE: To compare the surgical and functional results of full tendon rectus muscle transposition to the medial rectus muscle augmented with posterior medial fixation sutures in patients with adduction deficiency (ADD) and Duane's syndrome type 2. DESIGN: Retrospective nonconcurrent interventional comparative case series. PARTICIPANTS: Twelve consecutive patients with ADD and Duane's syndrome type 2. INTERVENTIONS: Six patients treated by the full tendon rectus muscle transposition procedure alone were compared with 6 patients treated by the full tendon rectus muscle transposition combined with posterior medial fixation sutures. MAIN OUTCOME MEASURES: Postoperative ocular alignment, duction improvement, binocular functions, and reoperation. RESULTS: In the augmented surgery group, mean distance and near deviation decreased by 74.5% and 74%, respectively, versus 56% and 59%, respectively, in the control group (P = 0.007 and 0.02, between-group comparison for distance and near deviation, respectively). Mean duction improved in 73% of the study group compared with 52% of controls (P = 0.025). No postoperative complications or duction anomalies were observed during follow-up in the entire cohort; reoperation was needed in 1 patient in the study group and in all 6 patients in the control group. CONCLUSIONS: The augmented full vertical rectus muscle procedure is a beneficial surgical approach for patients with ADD and Duane's retraction syndrome type 2, reducing the need for multiple extraocular muscle surgery. PMID- 15878080 TI - Tonometer accuracy. PMID- 15878082 TI - Endophthalmitis in an Asian population. PMID- 15878084 TI - Glaucoma detection. PMID- 15878086 TI - Visualizing the vitreous. PMID- 15878088 TI - Rate of foveal reattachment. PMID- 15878089 TI - Retinal venous pulsation and glaucoma. PMID- 15878090 TI - Pupil dilatation methods. PMID- 15878093 TI - Radiotherapy and Oncology comes of age. PMID- 15878094 TI - The clinical radiobiology of breast cancer radiotherapy. PMID- 15878095 TI - Fractionation sensitivity and dose response of late adverse effects in the breast after radiotherapy for early breast cancer: long-term results of a randomised trial. AB - BACKGROUND AND PURPOSE: Unlike squamous carcinomas, breast adenocarcinoma may be as sensitive to fraction size as late dose-limiting normal tissues. If so, fewer larger fractions would be as safe and effective as regimens based on 2.0 Gy fractions. The first step is to test the effects of radiotherapy fractions >2.0 Gy on late normal tissue responses in the breast after tumour excision and radiotherapy for early breast cancer. PATIENTS AND METHODS: One thousand four hundred and ten women with T1-3 N0-1 M0 invasive breast cancer were randomised between 1986-98 into one of three radiotherapy regimens after local tumour excision of early stage breast cancer; 50 Gy in 25 fractions (F) vs two dose levels of a test schedule giving 39 or 42.9 Gy in 13 F over 5 weeks. Fraction sizes were 2.0, 3.0 and 3.3 Gy, respectively. The primary endpoint was late change in breast appearance compared to post-surgical appearance scored from annual photographs blinded to treatment allocation. Secondary endpoints included palpable breast induration (fibrosis) and ipsilateral tumour recurrence. RESULTS: After a minimum 5-year follow up, the risk of scoring any change in breast appearance after 50 Gy/25 F, 39 Gy/13 F and 42.9 Gy/13 F was 39.6, 30.3 and 45.7%, from which an alpha/beta value of 3.6 Gy (95% CI 1.8-5.4) is estimated. The alpha/beta value for palpable breast induration was 3.1 Gy (95% CI 1.8-4.4). CONCLUSIONS: An alpha/beta value of around 3 Gy for late normal tissue changes in the breast is derived from the estimated equivalence of 41.6 Gy in 13 fractions and 50 Gy in 25 fractions over 5 weeks, in line with trial predictions. PMID- 15878096 TI - TGFB1 polymorphisms are associated with risk of late normal tissue complications in the breast after radiotherapy for early breast cancer. AB - Recent studies suggest that normal tissue radiosensitivity is influenced by single nucleotide polymorphisms (SNPs) in certain genes. In order to seek a confirmation of these findings, this study investigated SNPs in genes TGFB1 (position -509, codon 10 and codon 25), SOD2 (codon 16), XRCC1 (codon 399), XRCC3 (codon 241), APEX (codon 148) and ATM (codon 1853) in 26 breast cancer patients with marked changes in breast appearance after radiotherapy and 26 matched controls. Statistically significant associations were found between the TGFB1 codon 10 Pro allele (P=0.005) as well as the TGFB1 position -509 T allele (P=0.018) and increased risk of altered breast appearance. No significant associations were found for the remaining SNPs. PMID- 15878097 TI - Evaluation of set-up deviations during the irradiation of patients suffering from breast cancer treated with two different techniques. AB - PURPOSE: To compare reproducibility of set-up for two different treatment techniques for external irradiation of the breast. METHODS AND MATERIALS: In total, the analysis comprised 56 pairs of portal and simulator films for 14 consecutive patients treated following breast conserving therapy and 98 pairs of portal and simulator films for 20 consecutive patients treated after mastectomy. For the first group the tangential field technique (TF technique) was used, for the second the inverse hockey stick technique (IHS technique). Evaluation of the treatment reproducibility was performed in terms of systematic and random error calculated for the whole groups, comparison of set-up accuracy by means of comparison of cumulative distribution of the length of the displacement vector. RESULTS: In the IHS and TF techniques for medial and lateral fields, displacement larger than 5 mm occurred in 28.3, 15.8 and 25.4%, respectively. For the IHS technique, the systematic errors for lateral and cranial-caudal direction were 1.9 and 1.7 mm, respectively (1 SD), the random errors for lateral and cranial caudal direction were 2.0 and 2.5 mm. For the TF technique, the systematic errors for ventral-dorsal and cranial-caudal direction were 2.6 and 1.3 mm for medial field and 3.7 and 0.7 mm for lateral fields, respectively, the random errors for lateral and cranial-caudal direction were 2.2 and 1.0 mm for medial field and 2.9 and 1.1 for lateral field, respectively. Rotations were negligible in the IHS technique. For the TF technique the systematic and random components amounted to about 2.0 degrees (1 SD). CONCLUSIONS: Both the inverse hockey stick and standard tangential techniques showed good reproducibility of patients' set-up with respect to cranial-caudal direction. For the TF technique, the accuracy should be improved for the medial field with respect to the ventral-dorsal direction. PMID- 15878098 TI - Reliability of portal control procedure in irradiation of breast cancer: a Bayesian analysis. AB - BACKGROUND AND PURPOSE: To determine whether the information gathered from a fixed number of periodic verification films accurately reflects the true imprecision in patient positioning during the whole radiation therapy of early breast cancer. PATIENTS AND METHODS: A total of 204 medial and lateral treatment fields were evaluated in 102 breast cancer patients treated with conservative surgery and radiation therapy. For each treatment field, the central lung distance was measured on portal films obtained from one simulation and four treatment controls at weekly intervals during breast irradiation. Systematic and random errors in patient positioning throughout all treatment fractions were estimated from the available controls using Bayesian methods. RESULTS: The average systematic and random errors during treatment controls were 2.7 and 1.9 mm, respectively. For these mean control values, the probabilities that the true systematic and random errors remain below 5mm during all treatment fractions were 99 and 100%, respectively. CONCLUSIONS: Reproducibility of patient positioning was supported by a virtually null probability for systematic or random errors greater than 5 mm during the whole radiation therapy. Weekly verification films seem to be sufficient to estimate patient positioning errors with high accuracy in radiotherapy of early breast cancer. PMID- 15878099 TI - A randomised trial of accelerated radiotherapy for localised invasive bladder cancer. AB - BACKGROUND AND PURPOSE: To evaluate the efficacy and toxicity of an accelerated fractionation regimen to treat localised muscle invasive bladder cancer. PATIENTS AND METHODS: A prospective randomised trial was undertaken in 229 patients randomised between 1988 and 1998 comparing accelerated fractionation (AF) to a dose of 60.8 Gy in 32 fractions over 26 days with conventional fractionation (CF) treating to 64 Gy in 32 fractions over 45 days. Accelerated fractionation was delivered using two fractions per day with a 6h gap between fractions and with the first daily fraction size being 1.8 Gy and the second daily fraction size being 2.0 Gy. There was a 1 week treatment gap after the first 12 fractions. Conventional fractionation was one fraction per day, 5 days per week. Eligible patients had clinical stage T2 or T3, N0 or N1, M0 transitional cell carcinoma. The primary endpoint of the trial was local control and the trial was powered to detect a 20% difference (alpha 0.05, power 90%). Secondary endpoints were toxicity and survival. RESULTS: In the initial phase of the trial, randomisation was unequal such that in total 129 patients were randomised to accelerated fractionation and 100 to conventional fractionation. Acute toxicity was evaluable in 121 patients treated with AF and 96 patients treated with CF. RTOG grade 2 or 3 bowel toxicity was noted in 44% of AF patients compared to 26% of CF patients (P trend =0.001). Acute grade 2 or 3 bladder toxicity was seen in 35% of AF patients compared to 36% of CF patients (P=0.99). Late radiation toxicity was evaluated in patients surviving free from local recurrence at 2 years post treatment. Late radiation toxicity equivalent to RTOG grade 2 or more had occurred in 44% (95% CI 34-55%) of AF patients and in 38% (95% CI 26-49%) of CF patients (logrank over 5 years follow-up P=0.23). There was no significant difference in analysis of time to loss of local tumour control comparing the two treatment arms; local recurrence was recorded in 29 of the 100 patients treated with CF and in 41 of 129 patients treated with AF (logrank P=0.86). There was also no significant difference between the treatment arms comparing disease-free survival and overall survival. The overall survival figures at 3 years were for AF 54% (95% CI 45-63%) and for CF 47% (95% CI 36-57%). By 5 years the overall survival was 37% for AF and 40% for CF. There were two treatment related deaths, both on the AF arm of the trial. CONCLUSIONS: This accelerated fractionation schedule did not improve on the efficacy of conventional fractionation for patients with T2 and T3 bladder cancer and accelerated fractionation was associated with increased acute bowel reactions. PMID- 15878100 TI - Phase II study of radiochemotherapy with vinblastine in invasive bladder cancer. AB - Concurrent vinablastine-based radiochemotherapy was evaluated in 84 bladder cancer patients. It was effective in more than half: tumour-specific survival (51% 9-year), local control rate (55% 9-year). The drawback was the impaired function of the bladder (9-year prevalence SOMA G3-4 symptoms: 66%), indicating the need for treatment aimed at reducing chronic morbidity. PMID- 15878101 TI - Randomized trial of 8 Gy in 1 versus 20 Gy in 5 fractions of radiotherapy for neuropathic pain due to bone metastases (Trans-Tasman Radiation Oncology Group, TROG 96.05). AB - BACKGROUND AND PURPOSE: Despite numerous randomized trials investigating radiotherapy (RT) fractionation schedules for painful bone metastases, there are very few data on RT for bone metastases causing pain with a neuropathic component. The Trans-Tasman Radiation Oncology Group undertook a randomized trial comparing the efficacy of a single 8 Gy (8/1) with 20 Gy in 5 fractions (20/5) for this type of pain. MATERIALS AND METHODS: Eligible patients had radiological evidence of bone metastases from a known malignancy with no change in systemic therapy within 6 weeks before or anticipated within 4 weeks after RT, no other metastases along the distribution of the neuropathic pain and no clinical or radiological evidence of cord/cauda equina compression. All patients gave written informed consent. Primary endpoints were pain response within 2 months of commencement of RT and time to treatment failure (TTF). The hypothesis was that 8/1 is at least as effective as 20/5 and the planned sample size was 270 patients. RESULTS: Between February 1996 and December 2002, 272 patients were randomized (8/1:20/5=137:135) from 15 centres (Australia 11, New Zealand 3, UK 1). The commonest primary cancers were lung (31%), prostate (29%) and breast (8%); index sites were spine (89%), rib (9%), other (2%); 72% of patients were males and the median age was 67 (range 29-89). The median overall survival (95% CI) for all randomized patients was 4.8 mo (4.2-5.7 mo). The intention-to-treat overall response rates (95% CI) for 8/1 vs 20/5 were 53% (45-62%) vs 61% (53 70%), P=0.18. Corresponding figures for complete response were 26% (18-34%) vs 27% (19-35%), P=0.89. The estimated median TTFs (95% CI) were 2.4 mo (2.0-3.3 mo) vs 3.7 mo (3.1-5.9 mo) respectively. The hazard ratio (95% CI) for the comparison of TTF curves was 1.35 (0.99-1.85), log-rank P=0.056. There were no statistically significant differences in the rates of re-treatment, cord compression or pathological fracture by arm. CONCLUSIONS: 8/1 was not shown to be as effective as 20/5, nor was it statistically significantly worse. Outcomes were generally poorer for 8/1, although the quantitative differences were relatively small. PMID- 15878102 TI - Pain flare following external beam radiotherapy and meaningful change in pain scores in the treatment of bone metastases. AB - BACKGROUND AND PURPOSE: To examine the incidence of pain flare following external beam radiotherapy and to determine what constitutes a meaningful change in pain scores in the treatment of bone metastases. PATIENTS AND METHODS: Patients with bone metastases treated with external beam radiotherapy were asked to score their pain on a scale of 0-10 before the treatment (baseline), daily during the treatment and for 10 days after completion of external beam radiation. Pain flare was defined as a two-point increase from baseline pain in the pain scale of 0-10 with no decrease in analgesic intake or a 25% increase in analgesic intake employing daily oral morphine equivalent with no decrease in pain score. To distinguish pain flare from progression of pain, we required the pain score and analgesic intake to return back to baseline levels after the increase/flare. They were also asked to indicate if their pain changed during that time compared to pre-treatment level. The change in pain score was compared with patient perception. RESULTS: Eighty-eight patients were evaluated in this study. There were 49 male and 39 female patients with the median age of 70 years. Twelve of 88 patients (14%) had pain flare on day 1. The overall incidence of pain flare during the study period ranged from 2 to 16%. A total of 797 pain scorings were obtained. Patients perceived an improvement in pain when their self-reported pain score decreased by at least two points. CONCLUSIONS: Our study confirms the occurrence of pain flare following the external beam radiotherapy in the treatment of bone metastases. Further studies are required to predict who are at risk for flare. Appropriate measures can be taken to alleviate the pain flare. The finding in the meaningful change in pain scores supports the investigator defined partial response used in some clinical trials. PMID- 15878103 TI - Effectiveness and toxicity of single-fraction radiotherapy with 1 x 8 Gy for metastatic spinal cord compression. AB - Toxicity and functional outcome were evaluated in 199 MSCC-patients irradiated with 1 x 8 Gy. Motor function improved in 54/199 patients (27%). Patients who regained walking ability were 20/78 (26%). Long-term-survivors (> or =12 months) who needed re-irradiation for in-field-recurrence were 19/65 (29%). Acute toxicity was mild, late toxicity not observed. A randomised trial comparing single-fraction RT to multi-fraction RT is mandatory. PMID- 15878104 TI - Is the alpha-beta ratio of prostate cancer really low? A prospective, non randomized trial comparing standard and hyperfractionated conformal radiation therapy. AB - BACKGROUND AND PURPOSE: The objectives of the current study were to compare genito-urinary (GU) and gastro-intestinal (GI) toxicities as well as biochemical control (bRFS) in prostate cancer, utilizing conventional (2.0 Gy daily) (STD) or hyperfractionated (HFX) conformal irradiation (CRT). HFX (1.2 Gy BID) was chosen as a radiobiological method to try to reduce long term sequelae without compromising local control. PATIENTS AND METHODS: Three-hundred-and-seventy consecutive patients (pts) entered this prospective, non-randomized trial in the period January 1993-January 2003; 209 were treated with STD and 161 with HFX CRT. All were evaluable for acute toxicity analysis, 179 (STD) and 151 pts (HFX) being evaluable for late sequelae and bRFS analyses. Pt characteristics were not statistically different in the two groups. CRT consisted of a 4-field technique for prostate and/or pelvic nodes and a 5-field boost with rectal shielding. Median doses were 74 and 79.2 Gy for STD and HFX patients respectively, the latter dose being isoeffective for tumour control assuming alpha/beta=10 (EQD(2)=73.9 Gy). Median follow-up was 29.4 months (25.2 mos for STD; 37.7 mos for HFX; P<0.01). The two regimens were compared in terms of acute and late GU and GI toxicities and 5-year bRFS by univariate and multivariate analyses. RESULTS: Acute grade> or =2 GU toxicity was higher in the STD group (48.6% versus 37.3% in HFX, P=0.03), while no significant difference was found for acute GI toxicity. Late grade> or =2 GU and GI toxicities were lower in the HFX group (5 year actuarial rate: GU: 10.1% versus 20.3%, P=0.05; GI: 6.0% versus 10.6%, P=0.18). Five-year bRFS were 70% (+/-13.8%, 95% CI) and 82.6% (+/-7.2%) for STD and HFX, respectively (P=0.44); a trend favouring HFX was found in the subgroup of pts who did not receive hormonal therapy (5-year bRFS: 85.9%+/-12.4% versus 63.9%+/-23.8%, P=0.15). Multivariate analysis revealed only risk groups and age statistically related to bRFS but not fractionation regimen. Using the Nahum Chapman TLCP model and prostate parameter set, which includes hypoxia, the TLCPs are approximately equal for the two regimens, whereas assuming alpha/beta=1.5 and no hypoxia we obtain 73% for the STD group but only 36% for the HFX group. CONCLUSIONS: As expected from radiobiological considerations, HFX reduces GI and GU late toxicities. Concerning early bRFS, our clinical findings suggest that HFX is no less effective than STD when delivering an isoeffective (alpha/beta=10) dose. Despite the relatively short follow-up, this result appears to be inconsistent with a low alpha/beta ratio for prostate cancer. PMID- 15878105 TI - Testicular dose and hormonal changes after radiotherapy of rectal cancer. AB - BACKGROUND AND PURPOSE: To measure the dose received by the testicles during radiotherapy for rectal cancer and to determine the contribution of each field of the pelvic box and the relevance for hormonal status. MATERIALS AND METHODS: In 11 patients (mean age 55.2 years) testicular doses were measured with an ionisation chamber between 7 and 10 times during the course of pelvic radiotherapy (50 Gy) for rectal carcinoma. Before and several months after radiotherapy luteinizing hormone, follicle stimulating hormone and total testosterone serum levels were determined. RESULTS: The mean cumulative radiation exposure to the testicles was 3.56 Gy (0.7-8.4 Gy; 7.1% of the prescribed dose). Seventy-three percent received more than 2 Gy to the testicles. Fifty-eight percent of the measured dose was contributed by the p.a. field, 30% by the a.p. field and 12% by the lateral fields. Mean LH and FSH levels were significantly increased after therapy (350%/185% of the pre-treatment values), testosterone levels decreased to 78%. No correlation could be found between changes of hormones and doses to the testis, probably due to the low number of evaluated patients. CONCLUSIONS: Radiotherapy of rectal carcinoma causes significant damage to the testis, as shown by increased levels of gonadotropins after radiotherapy. Most of the gonadal dose is delivered by the p.a. field, due to the divergence of the p.a. beam towards the testicles. The reduction in testosterone level may be of clinical concern. Patients who will receive radiotherapy for rectal carcinoma must be instructed about a high risk of permanent infertility, and the risk of endocrine failure (hypogonadism). Larger studies are needed to establish the correlation between testicular radiation dose and hormonal changes in this group of patients. PMID- 15878106 TI - Enhanced response to radiotherapy in tumours deficient in the function of hypoxia inducible factor-1. AB - BACKGROUND AND PURPOSE: To test the hypothesis that deficiency in expression of the transcription factor, HIF-1, renders tumours more radioresponsive than HIF-1 proficient tumours. PATIENTS AND METHODS: Tumours comprising mouse hepatoma cells lacking HIF-1beta (and thereby HIF-1 function) were grown in nude mice and radiation-induced growth delay compared with that seen for wild-type tumours and tumours derived from HIF-1beta negative cells where HIF-1 function had been restored. RESULTS: The xenografts that lack HIF-1 activity take longer to establish their growth and are more radioresponsive than both parental xenografts and those with restored HIF-1 function. Pre-treatment of the HIF-1 deficient xenografts with the hypoxic radiosensitizer misonidazole, had little effect on radioresponse. In contrast this treatment radiosensitized the parental xenografts. In spite of this, no difference in oxygenation status was found between the tumour types as measured by Eppendorf O(2)-electrodes and by binding of the hypoxic cell marker NITP. Admixing wild type and HIF-1 deficient cells in the same tumour at ratios of 1 in 10 and 1 in 100 restores the growth of the mixed tumours to that of a 100% HIF-1 proficient cell population. However, when comparing the effects of radiation on the mixed tumours, radioresponsiveness is maintained in those tumours containing the high proportion of HIF-1 deficient cells. CONCLUSIONS: The differences in radioresponse do not correlate with tumour oxygenation, suggesting that the hypoxic cells within the HIF-1 deficient tumours do not contribute to the outcome of radiotherapy. Thus, hypoxia impacts on tumour radioresponsiveness not simply because of the physio-chemical mechanism of oxygen with radiation-induced radicals causing damage 'fixation', but also because hypoxia/HIF-1 promotes expression of genes that allow tumour cells to survive under these adverse conditions. Further, the results from the cell mixing experiments uncouple the growth promoting effects of HIF-1 and the underlying mechanism by which HIF-1 may increase radiation resistance in solid tumours. PMID- 15878107 TI - Effects of keratinocyte growth factor (palifermin) administration protocols on oral mucositis (mouse) induced by fractionated irradiation. AB - BACKGROUND AND PURPOSE: Aim of this study was to assess the impact of the administration protocol of palifermin on amelioration of oral mucositis after fractionated irradiation. MATERIALS AND METHODS: Mouse tongue ulceration was analysed as the clinically relevant endpoint. Daily fractionated irradiation (5 x 3 Gy/week, days 0 to +4, +7 to +11, with a weekend gap on days +5 and +6) was followed by graded test doses on day +14, i.e. after a second weekend gap. Palifermin (5 mg/kg) was injected subcutaneously. In the first series of experiments, the effect of three daily injections (days -3, -2 and -1) was compared with a single administration either on day -2 or -1; all animals received a further injection on day +4. In the second series, a single or three injections were given in the weekend gap between fractionated irradiation (days +5 to +6), with an additional administration on day +11. In a final protocol, single weekly injections of palifermin were given either on days -3, +4 and +11, days +4, +11 and +18, or on days -3, +4, +11 and +18. RESULTS: The ED50 (dose after which ulcer induction is expected in 50% of the mice) to single dose irradiation was 11.5+/-0.7 Gy. The ED50 for test irradiation after 10 x 3 Gy was 5.7+/-1.6 Gy. Palifermin administration before the start of fractionated irradiation and on day +4 increased the ED50 to 10-12 Gy, administration over the first weekend and on day +11 to 11-15 Gy. Administration over three consecutive weekends, starting on day -3 or day +4, increased the ED50 to 13.0+/-0.1 and 14.9+/-0.3 Gy. Single weekly KGF administrations over four weekends, including the weekend prior to and after completion of radiotherapy, showed no further increase in ED50. CONCLUSIONS: A single palifermin injection during the weekend gap before or during fractionated irradiation is as effective as three applications. Onset of the palifermin treatment during the first weekend gap between fractionated irradiation is more effective than during the weekend before radiotherapy. The effect of palifermin on oral mucositis can be increased by three weekly injections, while four injections do not yield a further increase in ED50. PMID- 15878108 TI - Intensity modulated radiation therapy with multileaf collimators of different leaf widths: a comparison of achievable dose distributions. AB - PURPOSE: A planning study to analyze the impact of different leaf widths on the achievable dose distributions with intensity modulated radiation therapy (IMRT). METHODS: Five patients (3 intra- and 2 extra-cranial) with projected planning target volume (PTV) sizes smaller than 10 cm by 10 cm were re-planned with four different multileaf collimators (MLC). Two internal collimators with an isocentric leaf width of 4 and 10 mm and two add-on collimators with an isocentric leaf width of 2.75 and were evaluated. The inverse treatment planning system KonRad (Siemens Medical Solutions) was used to create IMRT 'step & shoot' plans. For each patient the same arrangement of beams and the same parameters for the optimization were used for all MLCs. The beamlet size for all treatment plans was chosen to coincide with the leaf width of the respective MLC. To evaluate the treatment plans 3D dose distributions and dose volume histograms were analyzed. As indicators for the quality of the PTV dose distribution the minimum dose, maximum dose and the standard deviation were used. For the organs at risk (OAR) the equivalent uniform dose (EUD) was calculated. To measure the dose coverage of the PTV the volume (V(90)) that received doses higher than 90% of the prescribed dose was calculated where for the conformity the dose conformity index given by Baltas et al. was determined. RESULTS: The MLC with the smallest leaf width yields the best mean value of all five patients for the PTV coverage and for the conformity. For the MLCs with the same leaf width, the add-on MLC leads to superior treatment plans than the internal MLC. This is due to the sharper penumbra of the add-on MLC. The number of IMRT field segments to deliver increased by approximately a factor of two if 2. MLC leafs are used instead of the standard 10 mm leafs. In case of the para-spinal patients the EUD value for the spinal cord is only reduced slightly by using MLCs with leaf widths smaller than 5 mm. For the intra-cranial the EUD value for some organs improved with reduced leaf widths while for some organs the 10 mm MLC leafs give comparable values. CONCLUSION: As expected the MLC with the smallest leaf width always yields the best PTV coverage. Reducing the leaf width from 4 to 2.75 mm results in a slight enhancement of the PTV coverage. With the selected organ parameters no significant improvement for most OAR was found. The disadvantage of the reduction of the leaf width is the increasing number of segments due to the more complex fluence patterns and therefore an increased delivery time. PMID- 15878109 TI - IMRT with the sliding window: comparison of the static and dynamic methods. Dosimetric and spectral analysis. AB - BACKGROUND AND PURPOSE: Aim of the study is the critical appraisal of the two delivery techniques for intensity-modulated treatments commonly known as dynamic and static step and shoot, in the framework of a sliding window multileaf sequencing method. METHODS: The study was performed using the solution commercialised by Varian with the Eclipse treatment planning system (TPS) and the Clinac accelerator. For a set of intensity modulated fluences, the calculated dose maps and the dose delivered to films were compared for the static and dynamic modes to verify the capability of the TPS to accurately model both the delivery modes. For these investigations, the gamma concept of Low et al. [Low D, Harms W, Mutic S, Purdy J. A technique for the quantitative evaluation of dose distributions. Med Phys 1998;25: 656-60] was applied demonstrating, in general, optimal modeling for both static and dynamic tests. Optimal and actual fluences, were analysed to ascertain the degree of the TPS accuracy in converting 'ideal' maps into realistic leaf motions. RESULTS: Among the methods used, the Webb's Modulation Index [Webb S. Use of a quantitative index of beam modulation to characterize dose conformality: illustration by a comparison of full beamlet IMRT, few-segment IMRT and conformal unmodulated radiotherapy. Phys Med Biol. 2003;48: 2051-2062] proved to be useful. Also, dose volume histogram analysis was applied on deliverable plans using more clinical tools. CONCLUSION: The dynamic delivery method seems to offer a more reliable agreement with the optimal calculations and, clinically, a slightly superior performance in terms of target coverage. PMID- 15878110 TI - A prospective study on radiation pneumonitis following conformal radiation therapy in non-small-cell lung cancer: clinical and dosimetric factors analysis. PMID- 15878111 TI - Monotherapy for stage T1-T2 prostate cancer: radical prostatectomy, external beam radiotherapy, or permanent seed implantation. PMID- 15878112 TI - Interaction of tetraphenyl-porphyrin derivatives with DPPC-liposomes: an EPR study. AB - The effect of the symmetry and polarity of the porphyrin molecules on their membrane localization and interaction with membrane lipids were investigated by electron paramagnetic resonance (EPR). For this purpose, two glycoconjugated tetraphenyl porphyrin derivatives were selected, respectively, symmetrically and asymmetrically substituted. Small unilamellar liposomes composed of dipalmitoylphosphatidylcholine (DPPC) and spin labeled stearic acids were prepared. The spin probe was located at the 5th or 7th or 12th or 16th position of the hydrocarbon chain in order to monitor various regions of the lipid bilayer. EPR spectra of porphyrin-free and porphyrin-bound liposomes were recorded at various temperatures below and above the phase transition temperature of DPPC. The effect on membrane fluidity proved to be stronger with the asymmetrical porphyrin derivative than with the symmetrical one. The rigidity increased when the spin label was near lipid head groups. The difference observed between control and porphyrin-treated samples when measured below the main lipid transition temperature disappeared at higher temperature. When the spin label was near the end of the hydrophobic tails, the symmetrical porphyrin derivative caused increase in fluidity, while the asymmetrical one slightly decreased it. To explain this phenomenon we propose that the asymmetrical derivative exerts a stronger ordering effect caused by its fluorophenyl group located at the level of the lipid heads, which is attenuated to the hydrophobic tails. The perturbing effect of the symmetric derivative could not lead to similar extent of ordering at the head groups and looses the hydrocarbon chains deeper in the membrane. PMID- 15878113 TI - Photoreduction behavior of cytochrome c by zinc porphyrin in lipid media. AB - To elucidate the role of cardiolipin (CL) on redox behavior of cytochrome c (cyt c (III)), the photoreduction of cyt c using the photosensitization of zinc tetraphenylporphyrin in presence of triethanolamine (TEOA) as a sacrificial electron-donating reagent in various lipid media were studied. The initial rate of cyt c (III) photoreduction in various lipid, CL, l-alpha-phosphatidic acid (PA), dimethyldipalmitoylammonium bromide (DMPA) and Triton X-100 media were 1.0, 0.73, 0.80 and 0.67 micromol dm-3 min-1, respectively. The cyt c (III) photoreduction rate slightly increased by the addition of CL. PMID- 15878114 TI - Spectral analysis of laser-induced ultraweak delayed luminescence in cultured normal and tumor human cells: temperature dependence. AB - The emission spectrum of ultraviolet A laser induced ultraweak delayed luminescence in cell cultures of mammalian cells depended on the temperature during irradiation and photonic measurements. A new method using a sophisticated photomultiplier system was developed in order to find differences between normal and tumor cells. The maximal peak of the emitted light for cultures measured at low temperature of 10 degrees C was near 510 nm in the green visible region while following irradiation at 32 degrees C this maximum was shifted to yellow-orange at 570 nm both in normal and melanoma cells. Overall, this ultraweak photonic model of cultured cells provides to be a new and powerful non-invasive tool for developing new strategies in skin cancer detection. PMID- 15878115 TI - Evaluation of photolyase (Photosome) repair activity in human keratinocytes after a single dose of ultraviolet B irradiation using the comet assay. AB - Photosome is constituted of photolyases included in liposomes. Photolyase is a bacterial enzyme that can repair ultraviolet B (UVB)-induced cyclobutane pyrimidine dimers (CPD) in eukaryotic cells. A modified version of the alkaline comet assay has been set up to evaluate the repair activity of this enzyme after a single dose of UVB (312 nm, 0.06 J/cm2) in human keratinocytes. The formation of single strand breaks (SSB) induced by the UVA photoactivation of the enzyme (1.2 J/cm2) was inhibited by the pretreatment of the cells with 4 mM L ergothioneine (ERT) during 30 min at 37 degrees C. To increase the sensitivity of the comet assay, an additional lysis was used with a buffer containing sodium dodecyl sulfate (0.5%) and proteinase K (0.1 mg/ml) for 60 min at 37 degrees C. Unrepaired CPD by photolyase were revealed by a second enzymatic treatment with T4 endonuclease V, a CPD specific glycosylase. UVB irradiation increased the SSB level in keratinocytes and additional T4NV treatment enhanced this SSB level by 1.5-2.0-fold confirming that CPD were the major base modifications generated by UVB irradiation. UVA-photoactivated Photosome repaired CPD lesions and decreased the SSB levels by 2.6-3.3-fold. Photosome could be an additional component of sunscreens to reduce the development of skin cancer. PMID- 15878116 TI - Formation and reactivity of free radicals in 5-hydroxymethyl-2-furaldehyde--the effect on isoprenaline photostability. AB - Solutions of glucose are used as diluents for drugs in various drug infusions. When sterilized by heat small amounts of the substance 5-hydroxymethyl-2 furaldehyde (5-HMF) is produced from glucose. At a hospital ward such infusions may be exposed to irradiation; including UV-light. The photoreactivity of the furaldehyde is investigated. It is shown to photodestabilize the catecholamine isoprenaline. It is shown to be a producer, but also a consumer, of singlet oxygen. The excited triplet, cation and anion radical have been produced by pulse radiolysis and flash photolysis and their absorbance characteristics have been determined. The triplet absorption spectrum showed absorption bands at 320 and 430 nm with molar absorption coefficients of 4700 and 2600 M-1 cm-1, respectively. The anion radical showed absorption bands at 330 and 420 nm with molar absorption coefficients of 2000 and 300 M-1 cm-1, respectively. The cation radical had an absorption band at 320 nm with a molar absorption coefficient of 5000 M-1 cm-1. The quantum yield for the production of singlet oxygen, sensitized by the 5-HMF triplet, was determined to be 0.6, whilst the quantum yield for the triplet formation was 1.0. Aqueous solutions of 5-HMF were found to photoionize to yield the hydrated electron and the cation radical of 5-HMF in a biphotonic process. The influences of pH, buffer and glucose on the formation of transients were evaluated. The reactions between 5-HMF and the solvated electron, the hydroxyl radical and the superoxide were also studied. PMID- 15878117 TI - Reactive oxygen species produced upon photoexcitation of sunscreens containing titanium dioxide (an EPR study). AB - Commercial sunscreen products containing titanium dioxide were irradiated with lambda>300 nm and the formation of oxygen- (.OH, O2.-/.OOH) and carbon-centered radicals was monitored by EPR spectroscopy and spin trapping technique using 5,5 dimethyl-1-pyrroline N-oxide, alpha-phenyl-N-tert-butylnitrone (PBN), alpha-(4 pyridyl-1-oxide)-N-tert-butylnitrone as spin traps, and free nitroxide radical 4 hydroxy-2,2,6,6-tetramethylpiperidine N-oxyl. The photoinduced production of singlet oxygen was shown by 4-hydroxy-2,2,6,6-piperidine. The generation of reactive oxygen radical species upon irradiation of sunscreens significantly depends on their composition, as the additives present (antioxidants, radical scavengers, solvents) can transform the reactive radicals formed to less harmful products. The continuous in situ irradiation of titanium dioxide powder, recommended for cosmetic application, investigated in different solvents (water, dimethyl sulfoxide, isopropyl myristate) resulted in the generation of oxygen centered reactive radical species (superoxide anion radical, hydroxyl and alkoxyl radicals). PMID- 15878118 TI - Alterations of ciliate phosducin phosphorylation in Blepharisma japonicum cells. AB - We have previously reported that motile photophobic response in ciliate Blepharisma japonicum correlates with dephosphorylation of a cytosolic 28 kDa phosphoprotein (PP28) exhibiting properties similar to those of phosducin. Here we demonstrate in in vivo phosphorylation assay that the light-elicited dephosphorylation of the PP28 is significantly modified by cell incubation with substances known to modulate protein phosphatase and kinase activities. Immunoblot analyses showed that incubation of ciliates with okadaic acid and calyculin A, potent inhibitors of type 1 or 2A protein phosphatases, distinctly increased phosphorylation of PP28 in dark-adapted cells and markedly weakened dephosphorylation of the ciliate phosducin following cell illumination. An enhancement of PP28 phosphorylation was also observed in dark-adapted ciliates exposed to 8-Br-cAMP and 8-Br-cGMP, slowly hydrolysable cyclic nucleotide analogs and 3-isobutyryl-1-methylxanthine (IBMX), a non-specific cyclic nucleotide phosphodiesterase (PDEs) inhibitor. Only slight changes in light-evoked dephosphorylation levels of PP28 were observed in cells treated with the cyclic nucleotide analogs and IBMX. Incubation of ciliates with H 89 or KT 5823, highly selective inhibitor of cAMP-dependent protein kinase (PKA) and cGMP-dependent protein kinase (PKG), respectively, decreased PP28 phosphorylation levels in dark adapted cells, whereas the extent of light-evoked dephosphorylation of the phosphoprotein was only slightly influenced. Cell treatment with higher Ca2+ concentration together with ionophore A23187 in culture medium resulted in marked increase in PP28 phosphorylation levels, while quite an opposite effect was observed in cells exposed to Ca2+ chelators, EGTA or BAPTA/AM as well as calmodulin antagonists, such as trifluoperazine (TFP), W-7 or calmidazolium. Light-dependent dephosphorylation was not considerably affected by these treatments. The experimental findings presented here suggest that an endogenous light-dependent protein kinase-phosphatase system may be engaged in the alteration of phosducin phosphorylation in ciliate B. japonicum thereby to modulate the cell motile photophobic behavior. PMID- 15878119 TI - Kinetic isotope effects in the photochemical reaction cycle of ion transporting retinal proteins. AB - The kinetics of the photochemical reaction cycle of the bacteriorhodopsin, pharaonis halorhodopsin and proteorhodopsin were determined in H2O and D2O at low and high pH, to get insight in the proton dependent steps of the transport process. While all the steps of the bacteriorhodopsin photocycle at normal pH exhibited a strong isotope effect, the proton uptake step of the photocycle, measured at high pH, became independent of deuterium exchange, making plausible that this step, at low proton concentration, becomes concentration dependent, not mobility dependent. The proton transporting photocycle of the proteorhodopsin at its normal pH (9.5) shows a marked deuterium effect, while at high pH (12.2) this effect almost totally disappears. It was shown earlier that the proton uptake step of the proteorhodopsin is at the rise of the N form. As the proton concentration decreases with rising pH this step becomes the rate limiting, proton concentration dependent step, hiding all the other isotope dependent components. In the case of halorhodopsin in all the chloride, nitrate and proton transporting conditions the photocycle was not strongly affected by the deuterium exchange. While in the cases of the first two ions this seems normal, the absence of the deuterium effect in the case of the proton transporting photocycle was a puzzle. The only plausible explanation is that in the presence of azide the halorhodopsin transports not the proton, but a negatively charged ion the OH-, the mass and mobility of which is only slightly influenced by the deuterium exchange. PMID- 15878120 TI - Reduced hydroperoxidase (HPI and HPII) activity in the Deltafur mutant contributes to increased sensitivity to UVA radiation in Escherichia coli. AB - In Escherichia coli, Deltafur (ferric uptake regulator) mutants are hypersensitive to various oxidative agents, including UVA radiation (400-315 nm). Studies suggest that UVA radiation mediates its biological effects on bacteria via oxidative mechanisms that lead to reactive oxygen species, including the superoxide anion radical (O2.-), hydroxyl radical (HO.), hydrogen peroxide (H2O2) and singlet oxygen (1O2). There is accumulating evidence that Fur may play an important role in the defense against UVA radiation. In addition to regulating almost all genes directly involved in iron acquisition, Fur also regulates the expression of manganese and iron superoxide dismutase (MnSOD, FeSOD), key enzymes in the defense against oxygen toxicity in E. coli. In Deltafur mutants, there is a complete absence of FeSOD. Previous results suggest that the native iron chelating agent, enterobactin, which exists in increased levels in Deltafur mutants, is an endogenous chromophore for UVA, releasing Fe2+ into the cytoplasm to catalyze the production of highly reactive hydroxyl radicals. We now report that the hypersensitivity of Deltafur mutants to UVA irradiation is associated with reduced hydroperoxidase I (HPI) and hydroperoxidase II (HPII) activity, and is associated with a decrease in the transcription of katE and katG genes. The observed decrease in HPII activity in Deltafur mutants is also associated with reduced rpoS gene transcription. This study provides additional evidence that the Fur gene product, in addition to its known regulatory effect on the expression of SOD and iron uptake mechanisms, also regulates HPI and HPII activity levels in E. coli. An H2O2-inducible antioxidant defense system leading to an increase in HPI activity, is unaltered in Deltafur mutants. PMID- 15878121 TI - Photodynamic therapy for periodontal diseases: state of the art. AB - BACKGROUND: Photodynamic killing of periodontopathogenic bacteria may be an alternative to the systemic application of antibacterial drugs used in the treatment of periodontal diseases. Even though the method is still in the experimental stage, increasing bacterial resistance problems may promote the introduction of photodynamic therapy (PDT) into periodontal practice. AIM: In this review a literature survey is given of PDT as seen from a periodontal perspective. METHODS: In this review, the present knowledge and experience of PDT is summarized. Literature data are presented on drawbacks of conventional antibiotics, the mechanism of PDT, bactericidal effects of PDT as well as results of clinical efforts. The future prospects of the method are discussed. RESULTS: The application of photosensitizing dyes and their excitation by visible light enables effective killing of periodontopathogens. Encouraging studies using PDT in periodontitis and in peri-implantitis are known. CONCLUSION: Even though PDT is still in experimental stages of development and testing, the method may be an adjunct to conventional antibacterial measures in periodontology. Clinical follow up studies are needed to confirm the efficacy of the procedure. PMID- 15878122 TI - Molecular phylogenetic evidence for the monophyly of Fritillaria and Lilium (Liliaceae; Liliales) and the infrageneric classification of Fritillaria. AB - We present phylogenetic analyses of 37 taxa of Fritillaria (Liliaceae), 15 species of Lilium, and several outgroup taxa from Liliaceae s.s. to investigate the generic delimitation of Fritillaria in relation to Lilium as well as infrageneric relationships within Fritillaria. We used DNA sequences from the maturase-coding plastid matK gene and the trnK intron, the intron of the ribosomal protein-coding rpl16 plastid gene, and the nuclear ribosomal internal transcribed spacers (ITS). Phylogenetic analysis using maximum parsimony defined Fritillaria and Lilium (the latter including Nomocharis) as sister taxa. Fritillaria sections Fritillaria and Liliorhiza are supported in part, and some of the most enigmatic species usually included in Fritillaria (sections Petilium and Theresia and the monotypic genus Korolkowia) are closely related. The results support the new classification of Fritillaria proposed by Rix. We postulate independent origins of the underground bulbils found in Fritillaria davidii and the remainder of subgenus Liliorhiza. PMID- 15878123 TI - Identifying optimal incomplete phylogenetic data sets from sequence databases. AB - We introduce a new method for identifying optimal incomplete data sets from large sequence databases based on the graph theoretic concept of alpha-quasi-bicliques. The quasi-biclique method searches large sequence databases to identify useful phylogenetic data sets with a specified amount of missing data while maintaining the necessary amount of overlap among genes and taxa. The utility of the quasi biclique method is demonstrated on large simulated sequence databases and on a data set of green plant sequences from GenBank. The quasi-biclique method greatly increases the taxon and gene sampling in the data sets while adding only a limited amount of missing data. Furthermore, under the conditions of the simulation, data sets with a limited amount of missing data often produce topologies nearly as accurate as those built from complete data sets. The quasi biclique method will be an effective tool for exploiting sequence databases for phylogenetic information and also may help identify critical sequences needed to build large phylogenetic data sets. PMID- 15878124 TI - Cenozoic biogeography and evolution in direct-developing frogs of Central America (Leptodactylidae: Eleutherodactylus) as inferred from a phylogenetic analysis of nuclear and mitochondrial genes. AB - We report the first phylogenetic analysis of DNA sequence data for the Central American component of the genus Eleutherodactylus (Anura: Leptodactylidae: Eleutherodactylinae), one of the most ubiquitous, diverse, and abundant components of the Neotropical amphibian fauna. We obtained DNA sequence data from 55 specimens representing 45 species. Sampling was focused on Central America, but also included Bolivia, Brazil, Jamaica, and the USA. We sequenced 1460 contiguous base pairs (bp) of the mitochondrial genome containing ND2 and five neighboring tRNA genes, plus 1300 bp of the c-myc nuclear gene. The resulting phylogenetic inferences were broadly concordant between data sets and among analytical methods. The subgenus Craugastor is monophyletic and its initial radiation was potentially rapid and adaptive. Within Craugastor, the earliest splits separate three northern Central American species groups, milesi, augusti, and alfredi, from a clade comprising the rest of Craugastor. Within the latter clade, the rhodopis group as formerly recognized comprises three deeply divergent clades that do not form a monophyletic group; we therefore restrict the content of the rhodopis group to one of two northern clades, and use new names for the other northern (mexicanus group) and one southern clade (bransfordii group). The new rhodopis and bransfordii groups together form the sister taxon to a clade comprising the biporcatus, fitzingeri, mexicanus, and rugulosus groups. We used a Bayesian MCMC approach together with geological and biogeographic assumptions to estimate divergence times from the combined DNA sequence data. Our results corroborated three independent dispersal events for the origins of Central American Eleutherodactylus: (1) an ancestor of Craugastor entered northern Central America from South American in the early Paleocene, (2) an ancestor of the subgenus Syrrhophus entered northern Central America from the Caribbean at the end of the Eocene, and (3) a wave of independent dispersal events from South America coincided with formation of the Isthmus of Panama during the Pliocene. We elevate the subgenus Craugastor to the genus rank. PMID- 15878125 TI - Mitochondrial phylogeny reveals differential modes of chromosomal evolution in the genus Tatera (Rodentia: Gerbillinae) in Africa. AB - The African gerbils of the genus Tatera are widespread and abundant throughout sub-Saharan Africa. There is still today a certain controversy concerning the taxonomy of these rodents and very few attempts have been made to assess their systematic relationships. The present paper introduces findings based on the partial sequences of cytochrome b (495 bp) and the 16S rRNA (469 bp) mitochondrial genes of six (T. robusta, T. nigricauda, T. vicina, T. leucogaster, T. valida, and T. kempi) species together with two additional taxa. We also report the karyotypes of T. vicina and T. leucogaster. We propose that T. vicina should be considered as a valid species and show the monophyly of the robusta species group, with the exclusion of T. leucogaster. Our results show there is a different chromosomal evolutionary pattern within the two major lineages, which is recognizable through molecular phylogenetics. One is characterized by karyotype stability and the other by a considerable number of chromosomal rearrangements. The lineage divergence coincides with the formation of the East African Rift. The processes that led to the origin of the East African species seem to be related to the subsequent climatic changes, which caused cyclic contraction and expansion of the savannah biomes. Furthermore, geological activities that characterized East Africa during Plio-Pleistocene may also have contributed to lineage divergence. PMID- 15878126 TI - Taxon sampling effects in molecular clock dating: an example from the African Restionaceae. AB - Three commonly used molecular dating methods for correction of variable rates (non-parametric rate smoothing, penalized likelihood, and Bayesian rate correction) as well as the assumption of a global molecular clock were tested for sensitivity to taxon sampling. The test dataset of 6854 basepairs for 300 terminals includes a nearly complete sample of the Restio-clade of the African Restionaceae (272 of the 288 species), as well as 26 outgroup species. Of this, nested subsets of 35, 51, 80, 120, 150, and the full 300 species were used. Molecular dating experiments with these datasets showed that all methods are sensitive to undersampling, but that this effect is more severe in analyses that use more extreme rate smoothing. Additionally, the undersampling effect is positively related to distance from the calibration node. The combined effect of undersampling and distance from the calibration node resulted in up to threefold differences in the age estimation of nodes from the same dataset with the same calibration point. We suggest that the most suitable methods are penalized likelihood and Bayesian when a global clock assumption has been rejected, as these methods are more successful at finding optimal levels of smoothing to correct for rate heterogeneity, and are less sensitive to undersampling. PMID- 15878127 TI - Multiple events of horizontal transfer of the Minos transposable element between Drosophila species. AB - In this study the Minos element was analyzed in 26 species of the repleta group and seven species of the saltans group of the genus Drosophila. The PCR and Southern blot analysis showed a wide occurrence of the Minos transposable element among species of the repleta and the saltans groups and also a low number of insertions in both genomes. Three different analyses, nucleotide divergence, historical associations, and comparisons between substitution rates (d(N) and d(S)) of Minos and Adh host gene sequences, suggest the occurrence of horizontal transfer between repleta and saltans species. These data reinforce and extend the Arca and Savakis [Genetica 108 (2000) 263] results and suggest five events of horizontal transfer to explain the present Minos distribution: between D. saltans and the ancestor of the mulleri and the mojavensis clusters; between D. hydei and the ancestor of the mulleri and the mojavensis clusters; between D. mojavensis and D. aldrichi; between D. buzzatii and D. serido; and between D. spenceri and D. emarginata. An alternative explanation would be that repeated events of horizontal transfer involving D. hydei, which is a cosmopolitan species that diverged from the others repleta species as long as 14Mya, could have spread Minos within the repleta group and to D. saltans. The data presented in this article support a model in which distribution of Minos transposon among Drosophila species is determined by horizontal transmission balanced by vertical inactivation and extinction. PMID- 15878128 TI - Molecular criteria for determining new hybrid species--an application to the Sonneratia hybrids. AB - The possible hybrid origin of new species can usually be corroborated by molecular means. Here, we suggest that the segregation patterns of the molecular markers be further analyzed. A true hybrid species should show the patterns under continuous breeding among its members, at least beyond the F2 generation. We applied the guidelines to the putative hybrid species of Sonneratia, a widespread mangrove genus, and concluded that all the observed hybrids in this genus are simple F1's. Thus, S. x gulngai and S. x hainanensis are not true hybrid species. The segregation patterns of molecular markers should be heeded in interpreting the existence of hybrid species. PMID- 15878129 TI - Both host-plant phylogeny and chemistry have shaped the African seed-beetle radiation. AB - For the last 40 years, many authors have attempted to characterize the main patterns of plant-insect evolutionary interactions and understand their causes. In the present work on African seed-beetles (Coleoptera: Bruchidae), we have performed a 10-year field work to sample seeds of more than 300 species of potential host-plants (from the family Fabaceae), to obtain bruchids by rearing. This seed sampling in the field was followed by the monitoring of adult emergences which gave us the opportunity to identify host-plant use accurately. Then, by using molecular phylogenetics (on a combined data set of four genes), we have investigated the relationships between host-plant preferences and insect phylogeny. Our objectives were to investigate the level of taxonomic conservatism in host-plant fidelity and host-plant chemistry. Our results indicate that phylogenetically related insects are associated with phylogenetically related host-plants but the phylogeny of the latter cannot alone explain the observed patterns. Major host shifts from Papilionoideae to Mimosoideae subfamilies have happened twice independently suggesting that feeding specialization on a given host-plant group is not always a dead end in seed-beetles. If host-plant taxonomy and chemistry in legumes generally provide consistent data, it appears that the nature of the seed secondary compounds may be the major factor driving the diversification of a large clade specializing on the subfamily Mimosoideae in which host-plant taxonomy is not consistent with chemical similarity. PMID- 15878130 TI - The phylogeography of the gecko Cyrtopodion kotschyi (Reptilia: Gekkonidae) in the Aegean archipelago. AB - Cyrtopodion kotschyi is a small gecko, widely distributed on the islands of the Aegean archipelago and the adjacent mainland. We unveiled the phylogeography of the species by using a portion of the cytochrome oxidase I mitochondrial DNA gene from 35 insular and mainland populations. The distinct geographic distribution of the major clades of the phylogenetic tree and its topology suggest a spatial and temporal sequence of phylogenetic separations that coincide with some major paleogeographical separations during the geological history of the Aegean and support a mainly vicariant pattern of differentiation. The separation times and 95% confidence limits among the different clades were estimated according to two different paleogeographical scenarios. The very high interpopulation genetic divergence (up to 20% uncorrected pairwise distances) and the better concordance between paleogeographical and phylogenetic separations for one of the scenarios suggest that species differentiation started during Miocene (about 10 Mya) due to the fragmentation of the united landmass of "Agais" that was Aegean at that time. PMID- 15878131 TI - Utility of nuclear DNA intron markers at lower taxonomic levels: phylogenetic resolution among nine Tragelaphus spp. AB - Phylogenetic relationships among the nine spiral-horn antelope species of the African bovid tribe Tragelaphini are controversial. In particular, mitochondrial DNA sequencing studies are not congruent with previous morphological investigations. To test the utility of nuclear DNA intron markers at lower taxonomic levels and to provide additional data pertinent to tragelaphid evolution, we sequenced four nuclear DNA segments (MGF, PRKCI, SPTBN, and THY) and combined these data with mitochondrial DNA sequences from three genes (cytochrome b, 12S rRNA, and 16S rRNA). Our molecular supermatrix comprised 4682 characters which were analyzed independently and in combination. Parsimony and model based phylogenetic analyses of the combined nuclear DNA data are congruent with those derived from the analysis of mitochondrial gene sequences. The corroboration between nuclear and mtDNA gene trees reject the possibility that genetic processes such as lineage sorting, gene duplication/deletion and hybrid speciation account for the conflict evident in the previously published phylogenies. It suggests rather that the morphological characters used to delimit the Tragelaphid species are subject to convergent evolution. Divergence times among species, calculated using a relaxed Bayesian molecular clock, are consistent with hypotheses proposing that climatic oscillations and their impact on habitats were the major forces driving speciation in the tribe Tragelaphini. PMID- 15878132 TI - Molecular phylogenetics and biogeography of Neotropical piping guans (Aves: Galliformes): Pipile Bonaparte, 1856 is synonym of Aburria Reichenbach, 1853. AB - The Cracidae are Neotropical galliform birds with 11 genera currently recognized. To investigate the questioned validity of Pipile Bonaparte, 1856 and the monotypic Aburria Reichenbach, 1853 as separate genera, we gathered data from 2727 bp of mitochondrial DNA (cytochrome b, ND2 and control region) and 151 osteological characters. Our phylogenetic analyses of DNA sequences indicated that Aburria aburri is embedded within Pipile. Also, genetic distances between Aburria and any Pipile species are equivalent to the distances estimated for other congeneric cracid species, which genus status is not doubtful. Although the osteological characters do not have phylogenetic signal to solve the phylogenetic relationships at species level, five synapomorphies were found for Aburria and Pipile. Therefore, we suggest that Pipile should be merged with Aburria, which is the oldest described genus. We estimated that speciation in this group occurred in the Plio-Pleistocene, concordant with other birds, primates and rodents that have similar geographic distribution, and proposed a diversification hypothesis based on the occurrence of sea transgressions and the formation of the Amazon Lagoon. Therefore, we conclude that these palaeogeographic events may have contributed to Neotropical taxa diversification to a greater extent than previously suspected. PMID- 15878133 TI - Molecular phylogeny of parabasalids inferred from small subunit rRNA sequences, with emphasis on the Hypermastigea. AB - Small subunit rRNA gene sequences were identified without cultivation from parabasalid symbionts of termites belonging to the hypermastigid orders Trichonymphida (the genera Hoplonympha, Staurojoenina, Teranympha, and Eucomonympha) and Spirotrichonymphida (Spirotrichonymphella), and from four yet unidentified parabasalid symbionts of the termite Incisitermes minor. All these new sequences were analyzed by Bayesian, likelihood, and parsimony methods in a broad phylogeny including all identified parabasalid sequences available in databases and some as yet unidentified sequences probably derived from hypermastigids. A salient point of our study focused on hypermastigids was the polyphyly of this class. We also noted a clear dichotomy between Trichonymphida and the other parabasalid taxa. However, this hypermastigid order was apparently polyphyletic, probably reflecting its morphological diversity. Among Trichonymphida, Teranympha (Teranymphidae) grouped together with the members of the family Eucomonymphidae, suggesting that its family status is ambiguous. The monophyletic lineage composed by Spirotrichonymphida exhibited a narrower branching pattern than Trichonymphida. The root of parabasalids was examined but could not be discerned accurately. PMID- 15878134 TI - Intraordinal phylogenetics of treeshrews (Mammalia: Scandentia) based on evidence from the mitochondrial 12S rRNA gene. AB - Despite their traditional and continuing prominence in studies of interordinal mammalian phylogenetics, treeshrews (order Scandentia) remain relatively unstudied with respect to their intraordinal relationships. At the same time, significant morphological variation among living treeshrews has been shown to have direct relevance to higher-level interpretations of character state change as reconstructed in traditional interordinal studies, which have often included only a single species of treeshrew. Therefore, the importance of resolving relationships among treeshrews extends well beyond a better understanding of patterns of diversification within the order. A recent review highlighted several shortcomings in published studies of treeshrew phylogenetics based on morphology. Here we present the first investigation of treeshrew phylogenetics based on DNA sequences, utilizing previously published sequences from the mitochondrial 12S rRNA gene and combining them with newly generated sequence data from 15 species. Parsimony, likelihood, and Bayesian analyses all strongly support a sister relationship between Ptilocercus and the remaining species, further substantiating its recent elevation to familial status. Dendrogale is consistently recovered as the next taxon to diverge, but relationships among the remaining taxa are poorly supported by these data. We provide evidence for a relatively rapid radiation within the genera Tupaia and Urogale, but limited resolution precludes more than a cursory interpretation of biogeographic patterns. PMID- 15878135 TI - Systematics and evolution of the subfamily Gerbillinae (Mammalia, Rodentia, Muridae). AB - Although they represent a quarter of the mammalian species, the evolutionary relationships among as well as within, the main murid lineages are still controversial. The subfamily Gerbillinae is no exception as previous studies based on morphological, karyotypical, and allozyme characters are highly incongruent. Here, we present the first molecular phylogeny for gerbils based on cytochrome b and 12S rRNA mitochondrial genes. Results are largely congruent between the two genes as well as with the concatenated data set with most of the nodes being well-supported. Based on the topologies retrieved here, we (1) propose the identification of three main clades, (2) support the split of Tatera genus into an Asian and an African group, the latter including Gerbillurus species, and (3) provide some evidence towards the inaccuracy of subgeneric divisions within both Gerbillus and Meriones. In addition, the sharp contrast between the genetic characters and morphological data sets suggest high levels of convergence, probably as a result of strong environmental constraints imposed on these rodents adapted to arid and semi-arid regions. Finally, molecular datings for the various cladogenetic events are in good agreement with the known gerbilline fossil record and support an African origin with subsequent migrations to Asia. PMID- 15878136 TI - Phylogenetic analyses of mtDNA sequences corroborate taxonomic designations based on cuticular hydrocarbons in subterranean termites. AB - Cuticular hydrocarbons (CHCs) are valuable characters for the analysis of cryptic insect species with few discernible morphological characters. Yet, their use in insect systematics, specifically in subterranean termites in the genus Reticulitermes (Isoptera: Rhinotermitidae), remains controversial. In this paper, we show that taxonomic designations in Reticulitermes from California (USA) suggested in light of differences among CHC phenotypes are corroborated by phylogenetic analyses using mtDNA sequences. Analyses based on CHC phenotypes and supported, in part, by behavioral and ecological differences have suggested the presence of more species than the two currently recognized: R. hesperus Banks and R. tibialis Banks. We analyze a 680 base pair fragment of the mitochondrial DNA cytochrome oxidase (COII) gene from 45 new (21 collection localities) and two previously recorded samples of Reticulitermes from California using parsimony and maximum likelihood methods. Both methods result in trees with highly similar topologies. Bootstrapping indicates support for six clades of Reticulitermes, and corroborates groupings based on cuticular hydrocarbons. One of the clades, R. hesperus, is already recognized in California, while four clades appear to be previously undescribed taxa. Although identification of the final clade is inconclusive, it includes a sample putatively identified as R. tibialis. Therefore, using phylogenetic analyses we corroborate chemical characters used to identify taxa, associate a chemical phenotype with a previously described species, and provide additional support for undescribed taxa of Reticulitermes. PMID- 15878137 TI - Testing the trend towards specialization in herbivore-host plant associations using a molecular phylogeny of Tomoplagia (Diptera: Tephritidae). AB - Herbivorous insects are abundant and diverse and insect-host plant associations tend to be specialized and evolutionarily conserved. Some authors suggested that generalist insect lineages tend to become specialists, with host specialization leading to an evolutionary dead-end for the parasite species. In this paper, we have examined this tendency using a phylogenetic tree of Tomoplagia (Diptera: Tephritidae), a parasite of asteracean plants. We have tested the trend towards specialization in different hierarchical degrees of host specialization. The topology of the tree, the inference of ancestral hosts, and the lack of directional evolution indicated that specialization does not correspond to a phylogenetic dead-end. Although most Tomoplagia species are restricted to a single host genus, specialization does not seem to limit further host range evolution. This work emphasizes the advantages of the use of different levels of specialization and the inclusion of occasional hosts to establish a more detailed scenario for the evolution of this kind of ecological association. PMID- 15878138 TI - Testing the sister-group relationship of the Rhodniini and Triatomini (Insecta: Hemiptera: Reduviidae: Triatominae). PMID- 15878139 TI - Intron retention identifies a malaria vector within the Anopheles (Nyssorhynchus) albitaris complex (Diptera: Culicidae). PMID- 15878140 TI - Extrapolations are the Achilles heel of risk assessment. AB - This Reflections article considers the problems associated with the various extrapolations that are required for the estimation of human cancer risks from exposure to environmental carcinogens at low doses. These include extrapolation between species (particularly rodent to human), from responses at high doses to those at low doses, and among different stages of life. Reductions in uncertainty in risk estimates are closely coupled to the ability to conduct reliable extrapolations. The best way forward appears to be the use of data on mechanisms of carcinogenesis to develop bioindicators of responses related to the pathway to tumor formation. Such an approach is proposed based on the phenotypes represented by the six acquired characteristics forming the Hanahan-Weinberg model for carcinogenesis (The Hallmarks of Cancer). In addition, approaches can be established that use the Hanahan-Weinberg model as the basis for the collection and/or analysis of microarray or similar data. The reduction in reliance on default options and safety factors in the risk assessment process is a real possibility. PMID- 15878141 TI - Review of the genotoxicity of styrene in humans. AB - Styrene (CAS No. 100-42-5) is an important industrial chemical for which positive results have been reported in in vitro and in vivo genotoxicity assays. Styrene exposed workers have been studied extensively over two decades for the induction of various types of genotoxic effects. The outcomes of these studies have been conflicting, and where positive responses have been reported, it has proved difficult to demonstrate clear relationships between levels of damage reported and exposure levels. In this review, we have assessed studies addressing mutagenicity (chromosome aberrations, micronuclei and gene mutations) and other endpoints (sister chromatid exchanges, DNA breaks and DNA adducts) using criteria derived from the IPCS guidelines for the conduct of human biomonitoring studies. Based on the re-evaluated outcomes, the data are not convincing that styrene induces gene mutations. The evidence for induction of clastogenicity in occupationally exposed workers is less clear, with a predominant lack of induction of micronuclei in different studies, but conflicting responses in chromosome aberration assays. The results of numerous studies on sister chromatid exchanges do not provide evidence of a clear positive response, despite these being induced in animals exposed to styrene at high concentrations. However, there is evidence that both DNA adducts and DNA single strand breaks are induced in styrene workers. These types of damage are considered indicative of exposure of the target cells and interaction with cellular DNA but do not necessarily result in heritable changes. There is evidence that the metabolism of styrene in humans is affected by genetic polymorphisms of metabolizing genes and that these polymorphisms affect the outcome of in vitro mutagenicity studies on styrene. Therefore, studies that have addressed the potential of this factor to affect in vivo responses were considered. To date, there are no consistent relationships between genetic polymorphisms and induction of genotoxicity by styrene in humans, but further work is warranted on larger samples. The analyses of individual studies, together with a consideration of dose-response relationships and the lack of a common profile of positive responses for the various endpoints in different studies, provide no clear evidence that styrene exposure in workers results in detectable levels of mutagenic damage. However, evidence of exposure to genotoxic metabolites is demonstrated by the formation of DNA adducts and strand breaks. PMID- 15878142 TI - TP53 mutations as biomarkers for cancer epidemiology in Latin America: current knowledge and perspectives. AB - Due to particular social and economical development, and to the impact of globalization of lifestyles, Latin America shows a superposition of cancers that are frequent in low resource countries (gastric, oesophageal squamous cell and cervical cancers) and high resource countries (cancers of breast, colon and rectum, lung and prostate). Latin America thus offers opportunities for investigating the impact on changing lifestyle patterns on the occurrence of cancer. At the molecular level, mutations in the tumor suppressor gene TP53 are common in many cancers and their distribution can be informative of the nature of the mutagenic mechanisms, thus giving clues to cancer etiology and molecular pathogenesis. However most of the data available are derived from studies in industrialized countries. In this review, we discuss current trends on cancer occurrence in Latin American countries, and we review the literature available on TP53 mutations and polymorphisms in patients from Latin America. Overall, a total of 285 mutations have been described in 1213 patients in 20 publications, representing 1.5% of the total number of mutations reported world-wide. Except for hematological cancers, TP53 mutation frequencies are similar to those reported in other regions of the world. The only tumor site presenting significant differences in mutation pattern as compared to other parts of the world is colon and rectum. However, this difference is based on a single study with 35 patients. Recently, a characteristic TP53 mutation at codon 337 (R337H) has been identified in the germline of children with adrenocortical carcinoma in Southern Brazil. Further and better focused analyses of TP53 mutation patterns in the context of epidemiological studies, should help to improve our understanding of cancer etiology in order to develop appropriate health policies and public health programs in Latin America. PMID- 15878143 TI - Analytical methods in environmental effects-directed investigations of effluents. AB - Effluent discharges are released into aquatic environments as complex mixtures for which there is commonly either no knowledge of the toxic components or a lack of understanding of how known toxicants interact with other effluent components. Effects-directed investigations consist of chemical extraction and iterative fractionation steps directed by a biological endpoint that is designed to permit the identification or characterization of the chemical classes or compounds in a complex mixture responsible for the observed biological activity. Our review of the literature on effects-directed analyses of effluents for non-mutagenic as well as mutagenic endpoints showed that common extraction and concentration methods have been used. Since the mid-1980s, the methods have evolved from the use of XAD resins to C18 solid-phase extraction (SPE). Blue cotton, blue rayon, and blue chitin have been used specifically for investigations of mutagenic activity where polycyclic compounds were involved or suspected. After isolation, subsequent fractionations have been accomplished using SPE or a high-pressure liquid chromatography (HPLC) system commonly fitted with a C18 reverse-phase column. Substances in active fractions are characterized by gas chromatography/mass spectrometry (GC-MS) and/or other spectrometric techniques for identification. LC-MS methods have been developed for difficult-to-analyze polar substances identified from effects-directed studies, but the potential for LC-MS to identify unknown polar compounds has yet to be fully realized. Salmonella-based assays (some miniaturized) have been coupled with fractionation methods for most studies aimed at identifying mutagenic fractions and chemical classes in mixtures. Effects-directed investigations of mutagens have focused mostly on drinking water and sewage, whereas extensive investigations of non mutagenic effects have also included runoff, pesticides, and pulp mill effluents. The success of effects-directed investigations should be based on a realistic initial objective of each project. Identification of chemical classes associated with the measured biological endpoint is frequently achievable; however, confirmation of individual compounds is much more difficult and not always a necessary goal of effects-directed chemical analysis. PMID- 15878144 TI - GHB-C rats: the control line of GHB-sensitive (GHB-S) and GHB-resistant (GHB-R) rats. AB - Gamma-hydroxybutyric acid (GHB)-sensitive (GHB-S) and GHB-resistant (GHB-R) rats have been selectively bred for their opposite sensitivity to the sedative/hypnotic effect of GHB. This opposite sensitivity has been found to generalize to the GABA(B) receptor agonist, baclofen. A control line [named GHB control (GHB-C)] has been derived from the foundation stock of GHB-S and GHB-R rats. GHB-C rats have been bred without any evaluation of their sensitivity to GHB. The experiments described here were designed to evaluate the sensitivity of GHB-C rats, from the 13th generation, to the sedative/hypnotic effect of GHB (1 g/kg, i.p.) and baclofen (20 mg/kg, i.p.). All measures (onset, sleep time and r = sleep time/onset) of sensitivity to GHB- and baclofen-induced sedation/hypnosis in GHB-C rats were significantly different from and intermediate to those recorded in GHB-S and GHB-R rats. Furthermore, these values were similar to those recorded in the foundation stock. These results suggest that GHB-C rats may constitute a valid control line for GHB-S and GHB-R rats, representing the "general population" from which GHB-S and GHB-R rats were derived. Furthermore, the relative equidistance of sensitivity to GHB- and baclofen-induced sedation/hypnosis of GHB-C rats from those of GHB-S and GHB-R rats suggests that genetic factors contributes to the development of both sensitivity in GHB-S rats and resistance in GHB-R rats. PMID- 15878145 TI - Differences in the magnitude of long-term potentiation produced by theta burst and high frequency stimulation protocols matched in stimulus number. AB - Theta-burst stimulation (TBS: four pulses at 100 Hz repeated with 200 ms inter burst-intervals) and another commonly used high-frequency stimulation protocol (HFS: 1 s burst of equally spaced pulses at 100 Hz) were compared for the magnitude of LTP produced in rat hippocampal slices. The total number of pulses applied during tetanus (TET) was either 40, 100, 200, or 300. In a conventional analysis of the last 10 min of the post-TET period, a two-way ANOVA revealed no difference either in LTP of the field excitatory post-synaptic potential (fEPSP) between TBS and HFS or differences across pulse number at 40, 100, or 200 pulses. At 300 pulses, there was a significant main effect by pulse number but not by protocol. A linear regression analysis showed that stimulation protocol accounted for only about 10% of the change in magnitude while pulse number contributed to 30% of the change. However, when an extended analysis of the same data was performed across the entire post-TET period with a repeated-measure ANOVA, a small but persistent increase in TBS over HFS at 200 pulses was significant. A difference between TBS and HFS at 300 pulses that occurred only during the early phase of LTP was also significant. These results suggest that, over a range of stimuli, the number of pulses in an induction protocol, rather than the pattern of stimulation, determines the magnitude of late phase LTP, while TBS produces greater potentiation than HFS in the early phase of LTP with higher TET number. PMID- 15878146 TI - Development of a rat model to assess the efficacy of the somatosensory-evoked potential as indicator of analgesia. AB - Drug-induced changes in somatosensory-evoked potentials (SEPs) are considered to reflect an altered nociceptive state. Therefore, the SEP is proposed to be a parameter of analgesic efficacy. However, at present, SEPs have not been studied in relation to animal pain. The present study aims to develop a rat model in which this relationship can be studied based on Pavlovian fear conditioning. Therefore, rats, implanted with epidural electro-encephalogram recording electrodes, were randomly assigned to either a paired or random-control group and subjected to an aversive-to-appetitive transfer paradigm. During the aversive phase, the SEP-stimulation paradigm (5 mA square wave pulses, n = 72, of 2 ms duration each, with a stimulus frequency of 0.5 Hz; total duration 144 s) was used as the unconditioned stimulus (US), while a tone (40 s, 1500 Hz, 85 dB sound pressure level) was used as the conditioned stimulus (CS). During the appetitive phase, the CS was presented paired to the presentation of a sugar pellet. When compared to the random-control group, the paired group showed significantly more freezing behavior and significantly less reward-directed behavior in response to the CS in the appetitive phase. In addition, SEPs were not significantly affected by fear conditioning. Based on these results, we conclude that the SEP stimulation paradigm can be successfully employed as a US in fear conditioning. In future studies, fear conditioning can be carried out under different levels of an analgesic regimen to allow the changes in SEP parameters to be compared to changes in fear-induced behavior making this model potentially useful to validate SEP parameters as indicators of analgesia. PMID- 15878147 TI - A brightness-area-product-based protocol for the quantitative assessment of antigen abundance in fluorescent immunohistochemistry. AB - A problem frequently facing researchers examining abundance of expression of a given antigen is measurement. When the antigen is confined to the nucleus, absolute numbers of nuclei or a percentage of nuclei expressing the antigen in a given region can be estimated. When the antigen is localized to cytoplasm, cytoplasmic organelles or processes or membranes, the assessment becomes more difficult. In these settings, an observer/experimenter may assign a density score but intra- and inter-observer agreement using a three-tiered system, and finer resolution than this, is unlikely to be reproducible. Digital image analysis provides an opportunity to minimize observer bias in quantification of immunohistochemical staining. Previously, reported digital methods have mostly employed chromogen-staining methods and often report mean image brightness. We report a method for quantitatively assessing and expressing abundance of expression of an antigen in neural tissue stained with immunofluorescent methods by determining the brightness-area-product (BAP). The described protocol utilizes simple to use commercially available software and calculates BAP rather than mean brightness as a measure more representative of antigen abundance and visual interpretation. Accordingly, we propose this protocol as a useful adjunct to observer interpretation of fluorescent immunohistochemistry and its application to assessment of antigen abundance for varying patterns of antigen localization. PMID- 15878148 TI - A sequential fluorescence method for neurotransmitter-specific retrograde tracing in the central nervous system of the rat; utilizing True Blue and immunohistochemistry in combination with computer-assisted photography. AB - Aiming to map the distribution of spinally projecting, hypothalamic neurons containing neuronal nitric oxide synthase (nNOS), True Blue (TB) is injected into the rat spinal cord. After survival times of 7-14 days the animals are anaesthetized and perfused transcardially with a solution containing paraformaldehyde and sucrose. After dissection, the injection site is further fixed for 4-8 h, cut in a cryostat, and documented by computer-assisted digital photography. The brain region of interest is fixed for 4 h, rinsed in phosphate buffer for 48 h, sectioned, and photographically documented utilizing filter settings for visualization of TB. The brain sections are then immunohistochemically processed using a primary antibody against nNOS and a Texas Red (TR)-labelled secondary antibody and once again photographically documented, now using filter settings for visualization of TB and TR, respectively. Utilizing the Photoshop program, the TB containing cells can then be exactly aligned and the presence of TB and/or TR fluorescence in the same cell bodies are evaluated. This method for neurotransmitter-specific retrograde tracing derives its high sensitivity from the optimization of fixation/rinsing parameters, the use of appropriate fluorophores, and sequential digital microphotography. PMID- 15878149 TI - Real time computation of in vivo drug levels during drug self-administration experiments. AB - A growing body of evidence suggests that the drug concentration in the effect compartment of the body is the major factor regulating self-administration behavior. A novel computer-based protocol was developed to facilitate studies on mechanisms of drug addiction by determining correlations between drug levels and behavior during multiple drug injections and infusions. The core of the system is a user's program written in Medstate Notation language (Med-Associates, Inc.), which runs the self-administration session (with MED-PC software and hardware, Med-Associates, Inc.) and calculates the levels of infused and/or injected drugs in real time during the session. From the comparison of classical exponential and simple linear models of first-order kinetics, it is concluded that exponential solutions for the appropriate differential equations may be replaced with linear equations if the cycle of computation is much shorter than the shortest half-life for the drug. The choice between particular computation equations depends on assumptions about the pharmacokinetics of the particular drug: (i) one-, two- or three-compartment model, (ii) zero-, first- or second-order process of elimination, (iii) the constants of distribution and elimination half-lives of the drug are known or can be reasonably assumed, (iv) dependence of the constants on the drug level, and (v) temporal stability of all parameters during the session. This method of drug level computation can be employed not only for self administration but also for other behavioral paradigms to advance pharmacokinetic/pharmacodynamic modeling. PMID- 15878150 TI - Therapeutic benefit of TH-engineered mesenchymal stem cells for Parkinson's disease. AB - The present study was designed to assess the potential of marrow stromal cells (MSCs) to deliver therapeutic genes to the brain and result in biologically significant functional recovery. The tyrosine hydroxylase (TH) gene was transfected to MSCs with an adeno-associated virus (AAV) vector. MSCs expressing TH gene were transplanted into the striatum of Parkinson's disease (PD) rat. The asymmetric rotation of these models after apomorphine administration was detected every week after transplantation. Six weeks after grafting, animals were sacrificed. Some brains were sectioned to do TH immunohistochemistry. The others were used to detect the dopamine levels by high-performance liquid chromatograph and electrochemical detection (HPLC-ECD). The results showed that MSCs multiply rapidly and formed fibroblast colony-forming units in primary culture. The gene expression efficiency was about 75%. The rounds of asymmetric rotation after apomorphine administration decreased after TH-engineered MSCs were grafted. Histological examination showed that TH gene was expressed around the transplantation points. The dopamine level in the lesioned striatum of rats injected with TH-MSCs was significantly greater than that in rats treated with LacZ-MSCs (P < 0.05). All the data demonstrated that MSCs could readily be genetically engineered. Therefore, MSCs could be useful gene delivery vehicles of gene therapy for Parkinson's disease. PMID- 15878151 TI - A modified method for generation of neural precursor cells from cultured mouse embryonic stem cells. AB - The pluripotency and high proliferative capacity of embryonic stem (ES) cells make them an attractive source of different cell types for biomedical research and cell replacement therapies. It has been demonstrated that ES cells can be induced into neural precursor cells (NPCs) under conditions. NPCs can be expanded in large numbers for significant periods of time to provide a reliable source of cells for transplantation in neurodegenerative disorders and injury of the central nervous system. This study describes a modified method for generation of NPCs from cultured mouse ES cells. PMID- 15878152 TI - The contribution of alcohol to night time crash risk and other risks of night driving. AB - Many studies show that driving at night is more risky in terms of crash involvements per distance travelled than driving during the day. The reasons for this include the more prevalent use of alcohol by drivers at night, the effects of fatigue on the driving task and the risk associated with reduced visibility. Although the consumption of alcohol prior to driving occurs most commonly at night, drink-driving is not inherently a night time risk factor. This study decomposes the New Zealand risk of driving at night into risk associated with alcohol and risk associated with inherently night time factors. The overall risk associated with alcohol use by drivers was shown to decrease with increasing age for the most risky situation analysed (male drivers on weekend nights). Given the levels of drinking and driving on weekend nights, the overall effect of alcohol was shown to contribute almost half of weekend night time risk for drivers aged under 40 on lower volume roads, but to contribute little to overall risk on higher-volume roads, consistent with other research showing that higher-volume roads are not favoured by drinking drivers. Risk at night relative to risk during the day (excluding risk associated with drinking and driving) was shown to decrease with age. Roads with illumination at night are less risky at night relative to during the day than roads without illumination. The risks estimated in this paper reflect the behaviour of the road users studied and their prevalence on the roads under the conditions analysed. PMID- 15878153 TI - The origins and spread of antimalarial drug resistance: lessons for policy makers. AB - Understanding the frequency with which new resistance alleles arise and their subsequent patterns of spread is critical to our attempts to manage drug resistance in parasite populations. We review recent molecular evolutionary studies utilizing marker loci situated close to resistance loci on the Plasmodium falciparum genome that have given surprising insights into the origins and spread of drug resistance loci. We discuss possible reasons for the patterns observed, and highlight the implications of these results for resistance management. In particular, we show that many resistance mutations have rather few independent origins. De novo mutation appears to be less important than migration for introducing resistance alleles into parasite populations. Attempts to manage drug resistance will be of limited effectiveness unless this is taken into account. PMID- 15878154 TI - Population biology and antimalarial resistance: The transmission of antimalarial drug resistance in Plasmodium falciparum. AB - Malaria morbidity and mortality continue to increase across sub-Saharan Africa. This is largely as a result of the continued use of chloroquine and sulfadoxine pyrimethamine, despite widespread resistance. Although eliminating the asexual stages of Plasmodium falciparum is the focus of treatment of individual symptomatic patients, at a population level, reducing the carriage of gametocytes - the sexual stage responsible for infection of the mosquito vector - is necessary to limit the transmission of malaria parasites and the spread of antimalarial resistance. The probability of a mosquito being infected depends on the prevalence, duration and density of viable gametocyte carriage in the human host, although additional humoral and leukocyte factors also affect transmissibility. There is a log-sigmoid relationship between gametocyte density in the patients' blood and infectivity to the mosquito. The infectivity and thus transmission potential associated with a particular antimalarial treatment can be characterised as a function of blood gametocyte density and time, summing these over the acute and all subsequent recrudescences of that infection. Gametocyte carriage and infectivity to mosquitoes is consistently higher in patients infected with drug resistant compared with drug sensitive malaria parasites. It is the ratio of transmission potential in drug resistant versus sensitive infections that drives the spread of resistance. Early access to highly effective antimalarial treatment reduces the risk of disease progression and limits gametocyte carriage. The remarkable spread of sulfadoxine-pyrimethamine (SP) resistance across vast regions results from the very high post-treatment prevalence and density of gametocyte carriage following SP treatment. In areas of low intensity malaria transmission, the gametocyte-reducing effect of widespread use of artemisinin-based combination therapy has resulted in a sustained decrease in malaria transmission and a decrease in the spread of resistance. Malaria treatment policy should be based primarily on therapeutic efficacy against asexual stages, but should also consider transmission reduction potential. Artemisinin-based combination therapies are the only antimalarials currently available which rapidly reduce both asexual and gametocyte stages of the P. falciparum lifecycle. PMID- 15878155 TI - Oligomeric behavior of Bordetella pertussis adenylate cyclase toxin in solution. AB - Adenylate cyclase (AC) toxin from Bordetella pertussis inserts into eukaryotic cells, producing intracellular cAMP, as well as hemolysis and cytotoxicity. Concentration dependence of hemolysis suggests oligomers as the functional unit and inactive deletion mutants permit partial restoration of intoxication and/or hemolysis, when added in pairs [M. Iwaki, A. Ullmann, P. Sebo, Mol. Microbiol. 17 (1995) 1015-1024], suggesting dimerization/oligomerization. Using affinity co precipitation and fluorescence resonance energy transfer (FRET), we demonstrate specific self-association of AC toxin molecules in solution. Flag-tagged AC toxin mixed with biotinylated-AC toxin, followed by streptavidin beads, yields both forms of the toxin. FRET measurements of toxin, labeled with different fluorophores, demonstrate association in solution, requiring post-translational acylation, but not calcium. AC toxin mixed with DeltaR, an inactive mutant, results in enhancement of hemolysis over that with wild type alone, suggesting that oligomers are functional. Dimers and perhaps higher molecular mass forms of AC toxin occur in solution in a manner that is relevant to toxin action. PMID- 15878156 TI - Local application of prostaglandin E2 reduces trap, calcitonin receptor and metalloproteinase-2 immunoreactivity in the rat periodontium. AB - It has been shown that prostaglandin E2 (PGE2) locally released adjacent to the mandible over a 20-day period increases alveolar bone area, in part, due to a reduction in the percentage of eroded surface. To determine the effect of PGE2 on alveolar bone resorption, left mandibles from 24 Lewis rats were treated over a 20-day period with a local application of PGE2 (0.1, 0.05 or 0.025 mg/day) or placebo. The right side served as the non-treated matched control. Tissue sections were stained for tartrate resistant acid phosphatase (TRAP) calcitonin receptor (CTR) and metalloproteinase-2 (MMP-2). Matched samples were analysed by Wilcoxon matched pairs test and, a non-parametric one-way analysis of variance compared groups of treatment. Those tissues treated with PGE2 at doses of 0.1 and 0.05 mg/day showed significantly reduced numbers of TRAP and CTR-positive multinucleated cells compared with matched controls (p<0.005), as well as significantly reduced numbers of TRAP- and CTR-positive multinucleated cells when compared with the placebo-treated group (p<0.001). The number of periodontal ligament cells expressing MMP-2 was also significantly reduced in tissues treated with the two higher doses of PGE2 (p<0.001) comparing with both matched controls and the placebo-treated group. Following a 20-day period, locally released PGE2 at doses of 0.1 and 0.05 mg/day appears to affect alveolar bone resorption in the periodontium of rats, as the number of multinucleated cells expressing TRAP and CTR are significantly reduced. Furthermore, the same doses of PGE2 also significantly reduced the expression of MMP-2 by the periodontal cells. PMID- 15878157 TI - The role of epidermal growth factor receptor in ethanol-mediated inhibition of activator protein-1 transactivation. AB - A potential mechanism underlying ethanol-induced alterations in gene expression is the disruption of transcription factor activity. Growth factor receptors, particularly receptor tyrosine kinases, play an important role in modulating many biological effects of ethanol. We demonstrated here that the expression of epidermal growth factor receptor (EGFR) mediated the effect of ethanol on the activity of transcription factor activator protein-1 (AP-1). Ethanol had little effect on AP-1 activity in the fibroblast cells devoid of EGFR (B82); however, it significantly suppressed AP-1 activity in B82 cells that were stably transfected with either a wild-type EGFR (B82L) or a kinase-deficient receptor (B82M721) in a concentration-dependent manner. EGF activated AP-1 only in B82L cells; the activation was mediated primarily by Akt and ERK. Ethanol inhibited EGF-induced EGFR autophosphorylation, phosphorylation of ERK as well as Akt and its substrate GSK-3beta, and subsequently blocked EGF-stimulated AP-1 activation in B82L cells. On the other hand, ethanol had little effect on EGF-stimulated JNK activation. Phorbol ester 12-O-teradecanoyl-phorbol-13-acetate (TPA) activated AP-1 in B82L and B82M721 cells, but not B82 cells. TPA-induced activation of ERK and PKCdelta was dependent on the expression of EGFR although the intrinsic kinase activity of EGFR was not required. In contrast, TPA-induced phosphorylation of p38 MAPK, JNKs and other PKC isoforms was independent of EGFR. Ethanol selectively inhibited TPA induced phosphorylation of ERK and PKCdelta, and modestly suppressed TPA stimulated AP-1 activation in B82L and B82M721 cells. Thus, EGFR plays a critical role in the interaction between ethanol and AP-1. PMID- 15878158 TI - Estrogenic protection against gp120 neurotoxicity: role of microglia. AB - HIV infection of the nervous system can cause neurotoxicity, and the glycoprotein gp120 of HIV seems to play a key role in this. gp120 is neurotoxic through a multi-cellular pathway, stimulating microglia to release excitotoxins, cytokines and reactive oxygen species, which then damage neurons. We have previously shown that estrogen decreases the neurotoxicity of gp120 in mixed neuronal/glial cultures. In this study, we determine whether estrogen a) decreases the collective neurotoxicity of the factors released by gp120-treated microglia, and/or b) enhances the ability of neurons to survive such factors. To do so, we established microglial cultures, mixed neuronal/glial hippocampal cultures, and neuron-enriched cultures, independently manipulating gp120 and estrogen exposure in each type of culture, and inducing neurotoxicity in neuron-containing cultures by introducing conditioned media from gp120-treated microglial cultures. We observe that estrogen can exert some small protective effects at the level of bolstering neuronal resistance, but that the bulk of its protective effects arise at the level of decreasing the neurotoxicity of factors released by microglia. PMID- 15878159 TI - Platelet-derived microparticles induce angiogenesis and stimulate post-ischemic revascularization. AB - OBJECTIVE: Platelet activation is accompanied by the release of microparticles. However, little is known about the role of platelet-derived microparticles (PMP) in the regulation of angiogenesis and related clinical situations. The aim of our study was to evaluate the effect of PMP on angiogenesis and to analyze its mechanisms. METHODS: Both in vitro (rat aortic ring model, cell invasion test) and in vivo (agarose bead transplantation, artificial cardiac ischemia in Sabra rats) approaches were used in the study. RESULTS: A dose-dependent pro-angiogenic effect of PMP was observed in the rat aortic ring model. This effect could be eliminated by inhibition of VEGF, bFGF, and PDGF, but not heparanase. PMP exerted their effect via PI 3-kinase, Src kinase, and ERK, whereas protein kinase C and p38 were not involved. Moreover, PMP induced invasion of endothelial cells through a layer of matrigel. This effect was mediated by VEGF, heparanase, and PDGF, but not bFGF. Furthermore, PMP induced angiogenesis in an in vivo model in which agarose beads containing PMP were transplanted subcutaneously into mice. In addition, the effect of PMP on angiogenesis was evaluated in the model of in vivo chronic myocardial ischemia in rats. Ischemia induced a decrease in the number of functioning capillaries (34+/-21.5 vs. 157+/-42.0 per view field), but their amount increased after injection of PMP into the myocarium (97+/-27.3; p<0.001 vs. ischemia without PMP). CONCLUSIONS: PMP induce angiogenesis both in vitro and in vivo. Injection of PMP into the ischemic myocardium might improve the process of revascularization after chronic ischemia. PMID- 15878160 TI - Do children have increased susceptibility for developing secondary acute myelogenous leukemia? AB - This study was undertaken to evaluate the effects of age on a child's susceptibility to developing leukemia following exposure to known leukemogenic agents. The clinical literature describing the risk of developing acute myelogenous leukemia (AML) following treatment with alkylating agents or topoisomerase reactive drugs (known leukemogens) was used as a basis for this investigation. Based on this preliminary assessment, the age of the child does not appear to be an independent variable for risk following treatment with either class of drug. Although the number of studies and cases was very small, the available scientific and medical literature does not support the hypothesis that children will necessarily have an altered susceptibility or increased risk of developing chemotherapy-induced AML. PMID- 15878161 TI - Resveratrol inhibits angiotensin II-induced endothelin-1 gene expression and subsequent proliferation in rat aortic smooth muscle cells. AB - Resveratrol is a phytoestrogen naturally found in grapes and is the major constituent of wine thought to have a cardioprotective effect. The aims of this study were to examine whether resveratrol alters angiotenisn II-induced cell proliferation and endothelin-1 gene expression and to identify the putative underlying signaling pathways in rat aortic smooth muscle cells. Cultured rat aortic smooth muscle cells were preincubated with resveratrol then stimulated with angiotensin II, after which [3H]thymidine incorporation and endothelin-1 gene expression were examined. The intracellular mechanism of resveratrol in cellular proliferation and endothelin-1 gene expression was elucidated by examining the phosphorylation level of angiotensin II-induced extracellular signal-regulated kinase (ERK). The inhibitory effects of resveratrol (1-100 microM) on angiotensin II-induced DNA synthesis and endothelin-1 gene expression were demonstrated with Northern blot and promoter activity assays. Measurements of 2'7'-dichlorofluorescin diacetate, a redox-senstive fluorescent dye, showed a resveratrol-mediated inhibition of intracellular reactive oxygen species generated by the effects of angiotensin II. The inductive properties of angiotensin II and H2O2 on ERK phosphorylation and activator protein-1-mediated reporter activity were found reversed with resveratrol and antioxidants such as N acetyl-cysteine. In summary, we speculate that resveratrol inhibits angiotensin II-induced cell proliferation and endothelin-1 gene expression, and does so in a manner which involves the disruption of the ERK pathway via attenuation of reactive oxygen species generation. Thus, this study provides important insight into the molecular pathways that may contribute to the proposed beneficial effects of resveratrol on the cardiovascular system. PMID- 15878162 TI - Icariin-mediated modulation of cell cycle and p53 during cardiomyocyte differentiation in embryonic stem cells. AB - The aim of this study was to investigate the possible inducible effects and to clarify the modulation by icariin of cell cycle and p53 expression in the differentiation of embryonic stem cells into cardiomyocytes in vitro. Embryonic stem cells were cultivated as embryoid bodies in hanging drops and induced to differentiate into cardiomyocytes by icariin at 10(-7) M. Cardiomyocytes were characterized by the expression of sarcomeric proteins, alpha-actinin and cardiac troponin T, by immunocytochemistry. Flow cytometry revealed that 10(-7) M icariin treatment for 48 h significantly induced the accumulation of cells in G0/G1 and reduced the proportion of cells in S phase. A marked increase in apoptosis rate was observed 48 h after icariin treatment. Icariin resulted in significantly increased expressions of p53 mRNA and protein, as determined by reverse transcription-polymerase chain reaction and Western blot analysis. During day 7+0 and 7+9 cardiac developmental stage, 10(-7) M icariin increased the level of p53 mRNA, but caused a parallel decrease in the level of p53 protein. In conclusion, icariin at 10(-7) M facilitated the directional differentiation of embryonic stem cells into cardiomyocytes. Results showed p53 to be an important regulator in the differentiation in embryonic stem cells treated with 10(-7) M icariin, controlling or adjusting the balance between differentiated cells and cells undergoing apoptosis. PMID- 15878163 TI - The death domain-associated protein modulates activity of the transcription co factor Skip/NcoA62. AB - Death domain-associated protein (Daxx) regulates both transcription and apoptosis. The role of Daxx in transcription is not well understood. Here, we show that Daxx interacts with Skip/NcoA62, a transcription cofactor that modulates the activity of oncoproteins including Ski and NotchIC. Daxx strongly binds with Skip both in vitro and in mammalian cells. This interaction is mediated by the PAH2 domain of Daxx and the highly conserved SNW domain of Skip. Daxx partially co-localizes with Skip in vivo and changes the cellular distribution of Skip. In addition, Skip represses transcription when tethered to a promoter, and Daxx antagonizes this activity. Furthermore, Skip is phosphorylated at serine 224 in its SNW domain. These results suggest a novel function of Daxx in transcription regulation through alteration of the cellular localization of Skip. PMID- 15878164 TI - Zinc ions selectively inhibit steps associated with binding and release of NADP(H) during turnover of proton-translocating transhydrogenase. AB - Transhydrogenase couples the redox reaction between NAD(H) and NADP(H) to proton translocation across a membrane. In membrane vesicles from Escherichia coli and Rhodospirillum rubrum, the transhydrogenase reaction (measured in the direction driving inward proton translocation) was inhibited by Zn(2+) and Cd(2+). However, depending on pH, the metal ions either had no effect on, or stimulated, "cyclic" transhydrogenation. They must, therefore, interfere specifically with steps involving binding/release of NADP(+)/NADPH: the steps thought to be associated with proton translocation. It is suggested that Zn(2+) and Cd(2+) bind in the proton-transfer pathway and block inter-conversion of states responsible for changing NADP(+)/NADPH binding energy. PMID- 15878165 TI - Lower extremity control and dynamics during backward angular impulse generation in forward translating tasks. AB - Observation of complex whole body movements suggests that the nervous system coordinates multiple operational subsystems using some type of hierarchical control. When comparing two forward translating tasks performed with and without backward angular impulse, we have learned that both trunk-leg coordination and reaction force-time characteristics are significantly different between tasks. This led us to hypothesize that differences in trunk-leg coordination and reaction force generation would induce between-task differences in the control of the lower extremity joints during impulse generation phase of the tasks. Eight highly skilled performers executed a series of forward jumps with and without backward rotation (reverse somersault and reverse timer, respectively). Sagittal plane kinematics, reaction forces, and electromyograms of lower extremity muscles were acquired during the take-off phase of both tasks. Lower extremity joint kinetics were calculated using inverse dynamics. The results demonstrated between task differences in the relative angles between the lower extremity segments and the net joint forces/reaction force and the joint angular velocity profiles. Significantly less knee extensor net joint moments and net joint moment work and greater hip extensor net joint moments and net joint moment work were observed during the push interval of the reverse somersault as compared to the reverse timer. Between-task differences in lower extremity joint kinetics were regulated by selectively activating the bi-articular muscles crossing the knee and hip. These results indicate that between-task differences in the control of the center of mass relative to the reaction force alters control and dynamics of the multijoint lower extremity subsystem. PMID- 15878166 TI - The nature of infant color categorization: evidence from eye movements on a target detection task. AB - Infants respond categorically to color. However, the nature of infants' categorical responding to color is unclear. The current study investigated two issues. First, is infants' categorical responding more absolute than adults' categorical responding? That is, can infants discriminate two stimuli from the same color category? Second, is color categorization in infants truly perceptual? Color categorization was tested by recording adults' and infants' eye movements on a target detection task. In Experiment 1, adults were faster at fixating a colored target when it was presented on a colored background from a different color category (between-category) than when it was presented on a colored background from the same color category (within-category), even when within- and between-category chromatic differences were equated in CIE (Committee International d'Eclairage) color space. This category effect was found for two chromatic separation sizes. In Experiment 2, 4-month-olds also responded categorically on the task. Infants were able to fixate the target when the background color was from the same category. However, as with adults, infants were faster at fixating the target when the target background chromatic difference was between-category than when it was within-category. This implies that infant color categorization, like adult color categorization, is truly perceptual. PMID- 15878167 TI - Pre-event discussion and recall of a novel event: how are children best prepared? AB - We investigated the conditions under which information 1 day before a novel event influenced 6-year-olds' recall 2 weeks later. In Experiment 1A, four preparation conditions included either the event goals, goals with narration of the event actions and objects, photographs of the objects with goals and narration, or photographs with narration and child verbalization of the actions and objects. Compared with an irrelevant preparation control condition, goals with narration reduced errors, but correct recall was increased only when photographs were included. Child verbalization together with goals, narration, and photographs increased correct recall relative to goals with narration. In Experiment 1B, neither photographs nor preparation alone improved recall relative to the control condition. Experiment 2 found no further advantage of increasing children's participation in the preparation via questions. Overall, the impact of preevent information on memory is influenced by the nature of the information and the child's participation in the preparation. PMID- 15878168 TI - KChIP2 modulates the cell surface expression of Kv 1.5-encoded K(+) channels. AB - The Kv channel interacting proteins (KChIPs) were identified in a yeast two hybrid screen using the N terminus of Kv4.3 as bait. Previous studies have demonstrated that KChIP2 associates with voltage-gated K(+) (Kv) pore-forming (alpha) subunits of the Kv4 subfamily and contributes to the formation of the rapidly inactivating and recovering Kv4-encoded cardiac transient outward K(+) channels, I(to,f). Here, we report that co-expression of KChIP2 (or KChIP1) also modulates the functional cell surface expression of Kv1.5-encoded K(+) channels in transiently transfected HEK-293 cells. In contrast to the effects of KChIP2 on Kv4 channels, however, co-expression of KChIP2 (or KChIP1) decreases Kv1.5 encoded K(+) currents. Although current densities are reduced, KChIP2 (or KChIP1) co-expression does not affect the time- or voltage-dependent properties of heterologously expressed Kv1.5-encoded K(+) currents. Immunohistochemical and cell surface biotinylation experiments demonstrate that KChIP2 reduces the cell surface expression of Kv1.5, likely by inhibiting forward trafficking from the endoplasmic reticulum. In addition, biochemical experiments reveal that KChIP2 co immunoprecipitates with Kv1.5 (as well as Kv4.2/Kv4.3) from adult mouse ventricles, demonstrating that, similar to other Kv accessory subunits, KChIP2 is a multifunctional Kv channel accessory subunit. Taken together, the results here suggest that KChIP2 contributes to the formation of functional mouse ventricular (Kv1.5-encoded) I(K,slow1) channels as well, perhaps, as other Kv1.5-encoded K(+) currents, including I(Kur) (I(K,ultrarapid)), in human atria. PMID- 15878169 TI - Lonesome Kv4.2 channels. PMID- 15878170 TI - Short-term caloric restriction improves ischemic tolerance independent of opening of ATP-sensitive K+ channels in both young and aged hearts. AB - Ischemic tolerance decreases with aging and the cardioprotective effect of ischemic preconditioning (IPC) is impaired in aged animals. Although lifelong caloric restriction (CR) profoundly affects the physiological and pathophysiological modifications induced by aging and markedly increases life span in several species, it is unclear whether short-term CR affects ischemic tolerance and IPC in aged hearts. Six-month-old (Y) and 24-month-old (O) Fischer 344 male rats were randomly divided into two groups; AL rats were fed ad libitum, whereas CR rats were fed 90% of the caloric intake of AL for 2 weeks followed by 65% of the caloric intake for 2 weeks. Isolated perfused hearts were subjected to 25 min of ischemia followed by 30 min of reperfusion with or without IPC. The recovery of LV function after reperfusion improved with IPC in ALY but not in ALO. CR improved the recovery of LV function in both CRY and CRO but the cardioprotective effect of IPC was not additive to that of CR. Neither 5 hydroxydecanoate nor glibenclamide abrogated the protective effect of CR in either CRY or CRO. The recovery of myocardial high-energy phosphates after reperfusion was better with CR in both generations. There was no difference in myocardial expression levels of AMP-activated kinase (AMPK) but AMPK-alpha phosphorylated at Thr172 increased with CR in both Y and O. In conclusion, short term CR improves myocardial ischemic tolerance independent of the opening of KATP channels in both Y and O. CR-induced cardioprotection is associated with an increase in activated AMPK. PMID- 15878171 TI - Inhibition of protein kinase C reduces left ventricular fibrosis and dysfunction following myocardial infarction. AB - Despite current therapies, chronic heart failure (CHF) remains a major complication of myocardial infarction (MI). The pathological changes that follow MI extend to regions remote from the site of infarction (non-infarct zone, NIZ) where fibrosis is a prominent finding. Although the mechanisms underlying this adverse remodeling are incompletely understood, activation of protein kinase C has recently been implicated in its pathogenesis. MI was induced in Sprague Dawley rats by ligation of the left anterior descending coronary artery. One week post-MI, animals were randomized to receive the PKC-inhibitor, ruboxistaurin (LY333531) for 4 weeks, or no treatment. When compared with sham-operated animals, post-MI rats showed a 33+/-7% reduction in fractional shortening over a 4 weeks period, that was attenuated by treatment with ruboxistaurin (6+/-11%, P<0.05). Increased matrix deposition was noted in the NIZ, particularly in the subendocardial region of post-MI rats, in association with elevated expression of the profibrotic growth factor, transforming growth factor-beta. These findings were also significantly reduced by ruboxistaurin. PKC-inhibition with ruboxistaurin led to attenuation in both the pathological fibrosis and impaired cardiac function that follow experimental MI, suggesting a possible role for this agent in preventing post-infarction heart failure. PMID- 15878172 TI - From mutation to clinical presentation: mechanisms in the black box. PMID- 15878173 TI - Impaired SERCA function contributes to cardiomyocyte dysfunction in insulin resistant rats. AB - Ventricular dysfunction in type 2 diabetic patients is becoming apparent early after diagnosis of diabetes, but the cellular mechanisms contributing to this dysfunction are not well established. Our group has recently identified cardiomyocyte dysfunction in diet-induced insulin resistant rats that have not developed type 2 diabetes. The present investigation was designed to determine cellular mechanisms contributing to slowed cardiomyocyte relaxation in sucrose (SU)-fed rats. SU-feeding was used to induce whole-body insulin resistance. After 9-12 weeks on diet, isolated ventricular myocyte shortening/relengthening were slower in SU-fed adult male Wistar rats (42-63%) compared to starch (ST)-fed controls. Cytosolic Ca2+ removal attributable to Na+/Ca2+ exchange (NCX) and to sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) was evaluated with fluo-3/AM. Caffeine-releasable Ca2+ and cytosolic Ca2+ clearing through NCX were normal, whereas Ca2+ uptake by SERCA was significantly slower in SU myocytes (330+/-29 ms) compared to ST cells (253+/-16 ms). Protein levels for SERCA, NCX and phospholamban were not affected by SU-feeding. Manipulating intracellular Ca2+ with various positive inotropic interventions (e.g. post-rest potentiation, isoproterenol) and changes in stimulus frequency demonstrated that mechanical properties can be improved in subsets of myocytes. Thus, we conclude that impaired SERCA activity (with normal protein content) contributes to cardiomyocyte dysfunction in insulin resistant animals, whereas NCX function and expression are normal. These results suggest that subtle changes in Ca2+ regulation which occur prior to overt ventricular dysfunction/failure, may be common to early stages of a number of disorders involving insulin resistance (e.g. diabetes, obesity, syndrome X and hypertension). PMID- 15878174 TI - Structural basis for the high-affinity binding of nucleoporin Nup1p to the Saccharomyces cerevisiae importin-beta homologue, Kap95p. AB - Macromolecules are transported across the nuclear envelope most frequently by karyopherin/importin-beta superfamily members that are constructed from HEAT repeats. Transport of Kap95p (yeast importin-beta), the principal carrier for protein import, through nuclear pore complexes is facilitated by interactions with nucleoporins containing FG repeats. However, Nup1p interacts more strongly with Kap95p than other FG-nucleoporins. To establish the basis of this increased affinity, we determined the structure of Kap95p complexed with Nup1p residues 963 1076 that contain the high-affinity Kap95p binding site. Nup1p binds Kap95p at three sites between the outer A-helices of HEAT repeats 5, 6, 7 and 8. At each site, phenylalanine residues from Nup1p are buried in hydrophobic depressions between adjacent HEAT repeats. Although the Nup1p and generic FG-nucleoporin binding sites on Kap95p overlap, Nup1p binding differs markedly and has contributions from additional hydrophobic residues, together with interactions generated by the intimate contact of the linker between Nup1 residues 977-987 with Kap95p. The length and composition of this linker is crucial and suggests how differences in affinity for Kap95p both between and within FG-nucleoporins arise. PMID- 15878175 TI - Structural analysis of dispersin B, a biofilm-releasing glycoside hydrolase from the periodontopathogen Actinobacillus actinomycetemcomitans. AB - Bacteria in a biofilm are enmeshed in a self-synthesized extracellular polysaccharide matrix that holds the bacteria together in a mass and firmly attaches the bacterial mass to the underlying surface. A major component of the extracellular polysaccharide matrix in several phylogenetically diverse bacteria is PGA, a linear polymer of N-acetylglucosamine residues in beta(1,6)-linkage. PGA is produced by the Gram-negative periodontopathogen Actinobacillus actinomycetemcomitans as well as by the Gram-positive device-associated pathogen Staphylococcus epidermidis. We recently reported that A.actinomycetemcomitans produces a soluble glycoside hydrolase named dispersin B, which degrades PGA. Here, we present the crystal structure of dispersin B at 2.0A in complex with a glycerol and an acetate ion at the active site. The enzyme crystallizes in the orthorhombic space group C222(1) with cell dimensions a=41.02A, b=86.13A, c=185.77A. The core of the enzyme consists a (beta/alpha)(8) barrel topology similar to other beta-hexosaminidases but significant differences exist in the arrangement of loops hovering in the vicinity of the active site. The location and interactions of the glycerol and acetate moieties in conjunction with the sequence analysis suggest that dispersin B cleaves beta(1,6)-linked N acetylglucosamine polymer using a catalytic machinery similar to other family 20 hexosaminidases which cleave beta(1,4)-linked N-acetylglucosamine residues. PMID- 15878176 TI - A bacterial biosensor of endocrine modulators. AB - The nuclear hormone receptors comprise one of the largest classes of protein targets for drug discovery, as their function has been linked to a variety of serious diseases, including several forms of cancer. Identifying novel compounds with the ability to modulate the function of these targets could lead to the development of effective therapeutics. In vivo sensors of ligand binding have emerged as tools that can greatly accelerate the lead identification process, allowing new drugs to be discovered more rapidly and cheaply. In this work, a novel sensor of nuclear hormone binding has been developed in Escherichia coli by constructing a fusion of the ligand-binding domain of the human estrogen receptor with a thymidylate synthase enzyme (TS). Expression of this fusion protein in TS deficient bacterial cells resulted in growth phenotypes that were dependent on the presence of estrogen. Subsequent replacement of the estrogen receptor with the ligand-binding domain of the human thyroid hormone receptor led to specific thyroid hormone-enhanced growth that was insensitive to estrogen. This biosensor was then challenged with a small library of estrogen and thyroid hormone analogues, and it was observed that levels of cell growth correlate well with ligand-binding affinity. Remarkably, this simple biosensor was able to discriminate between agonistic and antagonistic activities, as combinations of estrogen agonists had an additive impact on cell growth, whereas known estrogen antagonists were found to neutralize agonist effects. This system constitutes a technique for facile selection of lead compounds with potential medical applications. PMID- 15878177 TI - Engineering the structural stability and functional properties of the GI domain into the intrinsically unfolded GII domain of the yeast linker histone Hho1p. AB - Yeast Hho1p contains two domains, GI and GII, that are homologous to the single globular domain of the linker histone H1 (GH1). We showed previously that the isolated GI and GII domains have different structural stabilities and functional properties. GI, like GH1 and the related GH5, is stably folded at low ionic strength (10 mM sodium phosphate) and gives strong protection of chromatosome length DNA ( approximately 166 bp) during micrococcal nuclease digestion of chromatin. GII is intrinsically unfolded in 10 mM sodium phosphate and gives weak chromatosome protection, but in 250 mM sodium phosphate has a structure very similar to that of GI as determined by NMR spectroscopy. We now show that the loop between helices II and III in GII is the cause of both its instability and its inability to confer strong chromatosome protection. A mutant GII, containing the loop of GI, termed GII-L, is stable in 10 mM sodium phosphate and is as effective as GI in chromatosome protection. Two GII mutants with selected mutations within the original loop were also slightly more stable than GII. In GII, two of the four basic residues conserved at the second DNA binding site ("site II") on the globular domain of canonical linker histones, and in GI, are absent. Introduction of the two "missing" site II basic residues into GII or GII L destabilised the protein and led to decreased chromatosome protection relative to the protein without the basic residues. In general, the ability to confer chromatosome protection in vitro is closely related to structural stability (the relative population of structured and unstructured states). We have determined the structure of GII-L by NMR spectroscopy. GII-L is very similar to GII folded in 250 mM sodium phosphate, with the exception of the substituted loop region, which, as in GI, contains a single helical turn. PMID- 15878178 TI - Suppression of multidrug-resistant HIV-1 reverse transcriptase primer unblocking activity by alpha-phosphate-modified thymidine analogues. AB - A dipeptide insertion between codons 69 and 70 together with the amino acid substitution T215Y in the reverse transcriptase (RT)-coding region of human immunodeficiency virus type 1 (HIV-1) strains are known to confer phenotypic resistance to zidovudine (AZT) and stavudine (d4T). Phenotypic resistance correlates with an increased ATP-dependent phosphorolytic activity. Nucleoside alpha-boranophosphate diastereoisomers derived from AZT and d4T were tested as substrates of a multidrug-resistant HIV-1 RT (designated as SS RT) bearing a Ser Ser insertion at codons 69-70 and other drug resistance-related mutations, in DNA polymerization assays and ATP-mediated excision reactions. Using pre-steady-state kinetics, we show that SS RT can incorporate both R(p) and S(p) diastereoisomers, although R(p) is the preferred isomer. Chirality at the internucleotidic linkage formed upon incorporation of nucleoside alpha-boranophosphate did not affect ATP mediated excision. As reported for AZT and d4T-terminated primers, substituting Thr, Asn or Ser for Tyr215 abrogates the ATP-dependent phosphorolytic activity on primers terminated with alpha-boranophosphate derivatives of thymidine analogues. However, unlike in the case of AZT, eliminating the dipeptide insertion in SS RT had no effect on the ATP-mediated excision of primers terminated with alpha boranophosphate derivatives of d4T. Studies with ATP analogues showed that exchanging a non-bridging oxygen atom at the gamma-phosphate group for sulfur causes a significant reduction of the ATP-dependent phosphorolytic activity of SS RT. Interestingly, SS RT's excision activity is completely eliminated upon phosphorothioate substitution at the 3' end of primers terminated with AZT. These results suggest that phosphorothioate derivatives of currently approved drugs could be useful against excision-proficient HIV-1 strains. PMID- 15878179 TI - Prevalence of major depressive disorder in the general population of South Korea. AB - INTRODUCTION: Previous epidemiological studies have reported a high prevalence of major depressive disorder (MDD) in North America and Western Europe. However, little information exists on MDD in Asian countries. This study investigates the prevalence of MDD and its characteristics in the general population of South Korea. METHODS: A representative sample of the South Korean general population composed of 3719 non-institutionalized individuals aged 15 years or older was interviewed by telephone using the Sleep-EVAL system. The participation rate was 91.4%. The interviews covered sociodemographic characteristics, health care utilization, physical illnesses and Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV) psychiatric disorders. RESULTS: A depressive mood, i.e., feeling sad, downcast, having the blues or having lost interest in things formerly pleasant was reported by 20.9% of the sample without significant difference between men and women and among age groups. DSM-IV MDD was found in 3.6% (95% CI: 3.0-4.2%) of the sample. The prevalence of MDD was comparable among age groups. Shift workers were more likely to have MDD than daytime workers. Factor significantly associated with MDD were: being a woman, being a light or heavy smoker, perceiving one's health as being average or poor, doing physical activities at least three times per week in the evening, having a BMI below 18.5 kg/m2 and perceiving one's life as being moderately or highly stressful. CONCLUSIONS: Prevalence of MDD in Korea is higher than what it was previously estimated to be two decades ago. The number of individuals seeking help for depression was very low, and only a small number of MDD subjects received appropriate treatment for their condition. PMID- 15878180 TI - On realizing shapes in the theory of RNA neutral networks. AB - It is known (Reidys et al., 1997b. Bull. Math. Biol. 59(2), 339-397) that for any two secondary structures S,S' there exists an RNA sequence compatible with both, and that this result does not extend to more than two secondary structures. Indeed, a simple formula for the number of RNA sequences compatible with secondary structures S,S' plays a role in the algorithms of Flamm et al. (2001. RNA 7, 254-265) and of Abfalter et al. (2003. Proceedings of the German Conference on Bioinformatics, ) to design an RNA switch. Here we show that a natural extension of this problem is NP-complete. Unless P=NP, there is no polynomial time algorithm, which when given secondary structures S1,...,S(k), for k4, determines the least number of positions, such that after removal of all base pairs incident to these positions there exists an RNA nucleotide sequence compatible with the given secondary structures. We also consider a restricted version of this problem with a "fixed maximum" number of possible stars and show that it has a simple polynomial time solution. PMID- 15878181 TI - Differentiating mechanisms of toxicity using global gene expression analysis in Saccharomyces cerevisiae. AB - Genotoxic stress triggers a variety of biological responses including the transcriptional activation of genes regulating DNA repair, cell survival and cell death. Genomic approaches, which monitor gene expressions across large numbers of genes, can serve as a powerful tool for exploring mechanisms of toxicity. Here, using five different agents, we investigated whether the analysis of genome-wide expression profiles in Saccharomyces cerevisiae could provide insights into mechanisms of genotoxicity versus cytotoxicity. To differentiate the genotoxic stress-associated expression signatures from that of a general cytotoxic stress, we compared gene expression profiles following the treatment with DNA-reactive (cisplatin, MMS, bleomycin) and DNA non-reactive (ethanol and sodium chloride) compounds. Although each of the tested chemicals produced a distinct gene expression profile, we were able to identify a gene expression signature consisting of a relatively small number of biologically relevant genes capable of differentiating genotoxic and cytotoxic stress. The gene set includes such upregulated genes as HUG1, ECM4 and previously uncharacterized gene, YLR297W in the genotoxic and GAP1, CGR1 in the cytotoxic group. Our results indicate the potential of gene expression profile analysis for elucidating mechanism of action of genotoxic agents. PMID- 15878182 TI - Experience with a category alters hemispheric asymmetries for the detection of anomalies. AB - Previous research with both brain-damaged and neurologically intact individuals suggests that the right cerebral hemisphere (RH) is superior to the left cerebral hemisphere (LH) at detecting anomalies in objects. The current research assesses whether experience with a category is necessary for this RH advantage to emerge. Participants were taught the diagnostic criteria necessary to categorize two fictitious species of animals ("Dleebs" and "Tazes"). After training, participants were given a test in which half of the items were congruent with the diagnostic rules and half of the items were incongruent. Participants were tested on two occasions-once after the initial training session and once after five training sessions. The results demonstrated that experience is required for the RH advantage for anomaly detection to occur. On the first test, reaction times were faster when items were presented to the LH. After 5 days of training, reaction times were faster when items were presented to the RH. This interaction could be due to the fact that participants reported analyzing the items in terms of a series of features during the initial test, but analyzed the items as a configural whole as experience with the category increased. PMID- 15878183 TI - Telling good from bad news: ADHD differentially affects processing of positive and negative feedback during guessing. AB - Neuroimaging studies on ADHD suggest abnormalities in brain regions associated with decision-making and reward processing such as the anterior cingulate cortex (ACC) and orbitofrontal cortex. Recently, event-related potential (ERP) studies demonstrated that the ACC is involved in processing feedback signals during guessing and gambling. The resulting negative deflection, the 'feedback-related negativity' (FRN) has been interpreted as reflecting an error in reward prediction. In the present study, ERPs elicited by positive and negative feedback were recorded in children with ADHD and normal controls during guessing. 'Correct' and 'incorrect' guesses resulted in respectively monetary gains and losses. The FRN amplitude to losses was more pronounced in the ADHD group than in normal controls. Positive and negative feedback differentially affected long latency components in the ERP waveforms of normal controls, but not ADHD children. These later deflections might be related to further emotional or strategic processing. The present findings suggest an enhanced sensitivity to unfavourable outcomes in children with ADHD, probably due to abnormalities in mesolimbic reward circuits. In addition, further processing, such as affective evaluation and the assessment of future consequences of the feedback signal seems to be altered in ADHD. These results may further help understanding the neural basis of decision-making deficits in ADHD. PMID- 15878184 TI - A cross-fostering analysis of the effects of PCB 77 on the maternal behavior of rats. AB - Polychlorinated biphenyls (PCBs) are environmental contaminants known to cause multiple behavioral and developmental problems in humans and animals. In rats, gestational exposure to the PCB congener 3, 4, 3', 4'-tetrachlorobiphenyl (PCB 77) affects the brain and behavior of the offspring as well as the maternal behavior of the dams. Whether the behavior of dams is affected by direct effects of the contaminant or indirectly by actions of the PCB on the developing offspring is not known. We investigated this question using a cross-fostering paradigm in which pregnant rats were exposed to either oil vehicle or 2 mg/kg of PCB 77 on gestational days 6 through 18, and then raised pups that had been exposed to either oil or PCB 77 during gestation. Maternal behavior was observed on postnatal days 1, 2, 4 and 6. Some of the effects on maternal behavior, including an increase in the frequency of nursing bouts and in the amount of maternal autogrooming, can be ascribed to prenatal exposure of the litters to the PCB. Other behavioral effects, including an increase in time on the nest and in the amount of pup grooming as well as a reduction in high-crouch nursing, appear to be due to both direct effects of the PCB on the dams and effects mediated by changes in the offspring. Our results show that exposure to PCB 77 can have complex effects on the behavioral interactions between the dams and their litters with a potential impact on the development of the offspring. PMID- 15878185 TI - Fatty acid metabolism as a target for obesity treatment. AB - Although metabolites and energy balance have long been known to play roles in the regulation of food intake, the potential role of fatty acid metabolism in this process has been considered only recently. Fatty acid synthase (FAS) catalyzes the condensation of acetyl-CoA and malonyl-CoA to generate long-chain fatty acids in the cytoplasm, while the breakdown of fatty acids (beta-oxidation) occurs in mitochondria and is regulated by carnitine palmitoyltransferase-1 (CPT-1), the rate-limiting step for the entry of fatty acids into the mitochondria. Inhibition of FAS using cerulenin or synthetic FAS inhibitors such as C75 reduces food intake and induces profound reversible weight loss. Subsequent studies reveal that C75 also stimulates CPT-1 and increases beta-oxidation. Hypotheses as to the mechanisms by which C75 and cerulenin mediate their effects have been proposed. Centrally, these compounds alter the expression profiles of feeding-related neuropeptides, often inhibiting the expression of orexigenic peptides. Whether through centrally mediated or peripheral mechanisms, C75 also increases energy consumption, which contributes to weight loss. In vitro and in vivo studies demonstrate that at least part of C75's effects is mediated by modulation of AMP activated protein kinase (AMPK), a known peripheral energy-sensing kinase. Collectively, these data suggest a role for fatty acid metabolism in the perception and regulation of energy balance. PMID- 15878186 TI - Detection of canine distemper virus by reverse transcriptase-polymerase chain reaction in the urine of dogs with clinical signs of distemper encephalitis. AB - In a prospective study we evaluated the use of the reverse transcriptase polymerase chain reaction (RT-PCR) in urine samples to diagnose canine distemper virus in dogs with progressive neurological disease. A fragment of the nucleoprotein gene of canine distemper virus was amplified from the urine of 22 distemper dogs. The body fluids and leukocytes of 12 asymptomatic dogs were RT PCR negative. RT-PCR of urine samples was more sensitive than serum and leukocytes and at least as sensitive as cerebrospinal fluid to screen for distemper in dogs with neurological signs and extraneural systemic signs. PMID- 15878187 TI - Crystal structures of 10alpha-gon-5-enes from the synthetic pathway to desogestrel. AB - X-ray crystallographic studies performed on the product of the ketalization reaction of 13beta-ethyl-11alpha-hydroxy-gon-5-ene-3,17-dione have lead to the unequivocal assignment of the 10alpha stereochemistry to C10, showing that an inversion of configuration occurred during formation of the 3,17-diketal. From the Swern oxidation of this compound, 11alpha-(methylthio)methoxy-10alpha-gonene was obtained as the major product instead of the desired 11-ketone. Modeling studies showed that the configurational instability at C10 is determined by the presence of the 11alpha-hydroxyl group. PMID- 15878188 TI - Development and analysis of a longitudinal and torsional type ultrasonic motor with two stators. AB - In this paper, a new longitudinal and torsional type ultrasonic motor with two stators is presented and investigated. Normally, such a motor consists of one rotor and one stator, and two types of transducers that are longitudinal PZT and torsional PZT are used to generate the desired elliptical locus on the stator surface. The operating frequency is at the resonance frequency of torsional transducer. In order to enhance the efficiency of the motor, however, the resonance frequencies of both transducers should be closed to each other. For the purpose of matching the resonance frequencies, a symmetrical structure is adopted in design of the motor. Furthermore, two rings are added to the stators in order to adjust the resonance frequencies of these two transducers. A finite element model is developed and ANSYS software is used to analyze the resonance frequencies of longitudinal vibration and torsional vibration as well as optimize the motor geometry. According to the FE results, an experimental prototype is fabricated and the experimental results agree well with the theoretical predictions. PMID- 15878189 TI - Electrochemical degradation of Acid Blue and Basic Brown dyes on Pb/PbO2 electrode in the presence of different conductive electrolyte and effect of various operating factors. AB - Electrocatalytic degradation of Acid Blue and Basic Brown dyes from simulated wastewater on lead dioxide anode was investigated in different conductive electrolytes. It was shown that complete degradation of these dyes is dependent primarily on type and concentration of the conductive electrolyte. The highest electrocatalytic activity was achieved in the presence of NaCl (2g/l) and could be attributed to indirect oxidation of the investigated dyes by the electrogenerated hypochlorite ions formed from the chloride oxidation. In addition, contribution from direct oxidation could also be possible via reaction of these organic compounds with the electrogenerated hydroxyl radicals adsorbed on the lead dioxide surface. In the presence of NaOH, the electrocatalytic activity of the employed anode was not comparable to that in NaCl due primarily to the absence of chloride. This indicates that dyes degradation in NaOH occurs exclusively via direct electrochemical process. However, in H2SO4, the electrode performance was poor due partially to the absence of chloride from the conductive solution. The possibility of electrode poisoning as a result of growth of adherent film on the anode surface or production of stable intermediates not easily further oxidized by direct electrolysis in H2SO4 might also be accountable for the poor performance observed in this conductive electrolyte. Optimizing the conditions that ensure effective electrochemical degradation of Acid Blue and Basic Brown dyes on lead dioxide electrode necessitates the control of all the operating factors. PMID- 15878190 TI - Decoloration of textile dyes by alginate-immobilized Trametes versicolor. AB - Alginate-immobilized Trametes versicolor decolorized Amaranth at similar rates in repeated batch culture when the dye was present in either (i) modified Kirk's medium containing 0.22 gl(-1) ammonium tartrate, (ii) the same buffer, thiamine, trace elements and glucose concentrations as in the modified Kirk's medium, or (iii) glucose alone at either 1, 5 or 10 gl(-1). With glucose alone (0.5 gl(-1)), Amaranth, Reactive Black 5, Reactive Blue 19 and Direct Black 22 had first-order decoloration rate constants of 0.56, 0.76, 0.52, and 0.15 h(-1), respectively. Mixtures of these dyes were also completely decolorized. After four successive decolorations, beads were kept in storage solutions for 48 d at 6 degrees C. CaCl2 (1g l(-1)) was the best storage solution as the beads were easier to handle and had the fastest decoloration rates after storage. Decoloration rates were faster with lower viscosity (less than 2000 cps) alginates and with softer beads which had a lower resistance to compression. Fungal colonization of the beads resulted in higher biomass concentrations with a corresponding higher decoloration rate but the beads became larger, had a lower resistance to compression and a higher percentage of bead breakage in a stirred tank reactor. Biomass, recovered from beads in which there was no growth, could be dispersed while the biomass from colonized beads formed a hollow, spherical shell due to growth on and near the bead surface and no growth in the bead interior. If alginate-immobilized T. versicolor is to be used in a stirred tank reactor, a high biomass loading during the immobilization phase and no fungal growth in the beads is recommended to have high decoloration rates and low bead breakage. PMID- 15878191 TI - Pepsinogen and H,K-ATPase mediate acid secretion in gastric glands of Triturus carnifex (Amphibia, Caudata). AB - The gastric glands of Triturus carnifex (Amphibia, Caudata) have been examined by histochemical and immunohistochemical methods with particular regard to hydrochloric acid and pepsinogen secretion. Fundic glands consist of mucous neck cells, endocrine cells and oxynticopeptic cells producing both pepsinogen and hydrochloric acid. The neck cells showed an unexpected distribution pattern which was only observed in the oral fundus, and produced neutral mucins with glycosidic residues of GalNAc and Gal beta1,3GalNAc, and in this respect they differ from the neck cells of anuran amphibians. The secretion of pepsinogen and hydrochloric acid as demonstrated by immunolabelling with anti-H,K-ATPase and with anti pepsinogen, respectively, seems not to vary significantly along the longitudinal axis of the stomach. The mechanism of gastric acid secretion seems to be mediated by an ATPase, having similar features to the mammalian gastric H,K-ATPase, and is localised in the luminal membrane and in the subapical cytoplasm of the oxynticopeptic cells. Unusually, the same cytoplasmic areas revealed binding specificity for the winged pea lectin (WPA) from Lotus tetragonolobus, even after beta elimination, indicating the presence of fucosyl residues in N-linked oligosaccharidic chains in glycoproteins of beta-H,K-ATPase subunits. PMID- 15878192 TI - Effect of semen collection method on pre- and post-thaw Guirra ram spermatozoa. AB - In this study, we evaluated the potential effect of the method of recovery (artificial vagina or electroejaculation) on the production and quality of Guirra ram spermatozoa cryopreserved for the possible constitution of a sperm bank. In order to address this question, we evaluated the effect of semen collection method on fresh semen quality parameters, including: volume, concentration, production, microscopic analysis (abnormal sperm and intact apical ridge) and sperm motility parameters determined by CASA system. For frozen-thawed semen, we evaluated motility parameters by CASA and intact apical ridge, acrosomal status, assessed by dual staining by IP and FITC-PNA and capacitation status, assessed by M540 and Yo-pro1, using flow cytometry. The main findings from this study were: (i) that electroejaculation resulted in a lower recovery efficiency (80% of the cases), as a consequence of contamination with urine or lack of response to the electrical stimulation; (ii) the fresh seminal quality was not significantly different between recovery methods, except for the concentration of spermatozoa, but total number of spermatozoa and the consequent number of possible seminal doses for artificial insemination were similar; and, (iii) a higher number of stable and functional spermatozoa (higher number of live non-capacitated cells, higher live acrosome intact cells and live acrosome reacted cells) were found for frozen-thawed spermatozoa collected by electro ejaculation than by artificial vagina. According to our results, we are able to develop both methodologies in the creation of the Guirra sperm bank. Assuming the advantages and limitations of both methodologies, in Guirra breed, would enable the rapid constitution of a sperm bank including samples from a large number of non-trained rams in a short period of time, which will increase the genetic variability, and so guarantee the conservation of this breed. PMID- 15878193 TI - Gene expression and developmental competence of bovine embryos produced in vitro under varying embryo density conditions. AB - The objective of this study was to determine the effects of oocyte/embryo density (embryo number:medium volume ratio) on developmental competence and relative transcription of developmentally important genes in in vitro-produced bovine embryos. Using standard procedures, bovine oocytes were matured, fertilized and cultured in vitro in groups of 5, 10, 20 or 30 oocytes/embryos per 100 microL medium drop (embryo number/medium volume ratios of 1:20, 1:10, 1:5 and 1:3.3, respectively). A semi-quantitative RT-PCR assay was used to determine the relative abundance of Hsp 70.1 and Glut-1 transcripts in Day-6 morulae, and in Day-7 and Day-8 blastocysts and expanded blastocysts. Cleavage rates (55.9, 49.1, 47.1 and 34.6%, respectively) and blastocyst developmental rates (24.1, 18.9, 20.4 and 12.6%, respectively) were higher in groups containing 5, 10 and 20 embryos versus 30 embryos per drop, but there were no significant differences among groups in hatching rates. The relative abundance of Hsp 70.1 transcripts in Day-7 blastocysts was higher in embryos produced in the 20-embryo than in the 5 embryo groups. There were no differences among embryos from all groups in the amount of Glut-1 mRNA (regardless of embryonic stage). However, there were differences in the pattern of Hsp 70.1 gene expression between distinct stages of embryo development and embryo density; these dissimilarities appeared to be associated with compromised development at higher embryo densities. In conclusion, the increase in embryo density during IVM, IVF, and IVC phases of in vitro embryo production might negatively affect embryo development and the pattern of gene expression of in vitro-produced bovine embryos. PMID- 15878194 TI - Bond strength of resin cements to H2O2-treated titanium plates. AB - OBJECTIVE: This study investigated the effects of H(2)O(2)-treatment of titanium surfaces on cement shear bond strengths, and characterized H(2)O(2)-treated titanium surfaces. METHODS: Using 34.5% hydrogen peroxide solution, cp Ti plates (10 mm x 10 mm x 1 mm) were treated by (1) an immersion method, and (2) halogen irradiation while immersed in H(2)O(2) for varying times. A cylindrical block (6 mm diameter, 4 mm height) of four different cements was bonded onto H(2)O(2) treated surfaces. The cement bond strengths were evaluated under shear mode. Treated surfaces were also characterized for color change, wettability, AC impedance, and transmission electron diffraction of stripped oxide film. RESULTS: The cement shear bond strength of cp Ti treated with H(2)O(2) and halogen for 160 s was the highest and was approximately 14 times higher than the un-treated control cp Ti plates. Bond strengths are correlated linearly to wettability. The more surface wetted with the cement material, the higher the resultant shear bond strength value. SIGNIFICANCE: These results suggest that a combined treatment of hydrogen peroxide and halogen light irradiation provides an effective surface condition with appropriate oxide film thickness to enhance the cement bond strength. PMID- 15878195 TI - Electron beam irradiation of dental composites. AB - OBJECTIVES: Electron beam irradiation can be used to influence the mechanical properties of polymers. It was the aim of this study to investigate whether dental composites can benefit from irradiation in order to achieve increased fracture toughness, work of fracture, hardness or less wear. METHODS: Two hundred rectangular specimens of five veneering composites were electron beam irradiated with 25, 100 and 200 kGy using an electron accelerator of 10 MeV. Fracture toughness, work of fracture, Vickers hardness, color changes and three-medium wear were measured and compared with non-irradiated specimens. RESULTS: Visible color changes (DeltaE>3) were observed with all composites and with all dose rates. Fracture toughness, work of fracture, Vickers hardness and resistance against wear increased significantly with few exceptions. Composites with a simple curing process needed higher dose rates while systems with a more complex curing procedure should be irradiated with lower dose rates. SIGNIFICANCE: Electron beam irradiation can significantly change the mechanical properties of dental composites. However, color changes can limit the use of irradiation for dentistry. PMID- 15878196 TI - Brief report: the Ego Identity Process Questionnaire: factor structure, reliability, and convergent validity in Dutch-speaking late adolescents. AB - The reliability and validity of a Dutch version of the Ego Identity Process Questionnaire (EIPQ) were evaluated. In Study 1, the instrument was found to exhibit a clear factor structure and acceptable reliability. Using Confirmatory Factor Analysis, both a model with two process factors (Commitment, Exploration) and a model with four content factors (Ideological Commitment, Ideological Exploration, Interpersonal Commitment, Interpersonal Exploration) evidenced a good fit once direction of wording was taken into account. In Study 2, moderate convergence was observed between Commitment and Exploration, and continuous measures of identity statuses and identity styles. The results of both studies combined indicated that the Dutch version of the EIPQ may be recommended as a research tool with college students. PMID- 15878197 TI - A fundamental approach for the estimation of the mechanical glass transition temperature in gelatin. AB - The paper constitutes an attempt to overcome the empiricism prevalent in the estimation of the glass transition temperature (Tg) of gelatin networks using rheological techniques. In doing so, it presents a study of the viscoelastic properties of a well-characterised gelatin sample covering the structural properties from the rubbery region to the glassy state. The pattern of oscillatory behaviour on shear is given by a master curve produced by shifting data obtained at different temperatures along the logarithmic time scale. Data reduction does not hold for all temperatures thus giving rise to thermorheological complexity. Within the temperature range at which molecular processes are represented by a simple distribution of relaxation times, a fundamental argument is developed to pinpoint the mechanical Tg. This should improve confidence in measured glassy properties over the empirical indicators found in the literature. As a demonstration, the glass transition temperature of gelatin at "zero moisture" obtained using the proposed framework of analysis is contrasted with earlier attempts to identify the mechanical Tg of gelatin solids. PMID- 15878198 TI - The effect of incorporating RGD adhesive peptide in polyethylene glycol diacrylate hydrogel on osteogenesis of bone marrow stromal cells. AB - Advances in tissue engineering require biofunctional scaffolds that can not only provide cells with structural support, but also interact with cells in a biological manner. To achieve this goal, a frequently used cell adhesion peptide Arg-Gly-Asp (RGD) was covalently incorporated into poly(ethylene glycol) diacrylate (PEODA) hydrogel and its dosage effect (0.025, 1.25 and 2.5 mm) on osteogenesis of marrow stromal cells in a three-dimensional environment was examined. Expression of bone-related markers, osteocalcin (OCN) and Alkaline phosphatase (ALP), increased significantly as the RGD concentration increased. Compared with no RGD, 2.5 mm RGD group showed a 1344% increase in ALP production and a 277% increase in OCN accumulation in the medium. RGD helped MSCs maintain cbfa-1 expression when shifted from a two-dimensional environment to a three dimensional environment. Soluble RGD was found to completely block the mineralization of marrow stromal cells, as manifested by quantitative calcium assay, phosphorus elemental analysis and Von Kossa staining. In conclusion, we have demonstrated that RGD-conjugated PEODA hydrogel promotes the osteogenesis of MSCs in a dosage-dependent manner, with 2.5 mm being optimal concentration. PMID- 15878199 TI - Primary pulmonary plasmacytoma involving bilateral lungs and marked hypergammaglobulinemia: differentiation from extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue. AB - A 71-year-old woman was referred to our hospital because of hyperproteinemia and serum M-protein (IgG-lambda type). Chest computed tomographic (CT) scan revealed a tumor in each lung and transbronchial lung biopsy was performed. Histopathological examination showed monotonous medullary proliferation of morphologically mature plasma cells. These cells were cIgG+, cIg-lambda+, CD 20+, CD 79 a+, CD 138+, cIg-kappa-, and CD3-. Since there were very few non-neoplastic plasma cells and small lymphocytes in addition to the absence of reactive lymph follicles and fibrosis, the patient was diagnosed as having plasmacytoma. There was no proliferation of plasma cells in the bone marrow. Thus, the lesion was finally characterized as primary pulmonary plasmacytoma. Treatment with melphalan/prednisolone resulted in considerable decrease in the serum IgG level and regression of the pulmonary tumors. The effectiveness of the chemotherapy could confirm our diagnosis, although MALT-type lymphoma with plasmacytic differentiation cannot be completely ruled out. PMID- 15878200 TI - Chromium toxicity in plants. AB - Due to its wide industrial use, chromium is considered a serious environmental pollutant. Contamination of soil and water by chromium (Cr) is of recent concern. Toxicity of Cr to plants depends on its valence state: Cr(VI) is highly toxic and mobile whereas Cr(III) is less toxic. Since plants lack a specific transport system for Cr, it is taken up by carriers of essential ions such as sulfate or iron. Toxic effects of Cr on plant growth and development include alterations in the germination process as well as in the growth of roots, stems and leaves, which may affect total dry matter production and yield. Cr also causes deleterious effects on plant physiological processes such as photosynthesis, water relations and mineral nutrition. Metabolic alterations by Cr exposure have also been described in plants either by a direct effect on enzymes or other metabolites or by its ability to generate reactive oxygen species which may cause oxidative stress. The potential of plants with the capacity to accumulate or to stabilize Cr compounds for bioremediation of Cr contamination has gained interest in recent years. PMID- 15878201 TI - Fusion protein of CDR mimetic peptide with Fc inhibit TNF-alpha induced cytotoxicity. AB - The variable regions of antibodies play central roles in the binding with antigens. Based on the model of a tumour necrosis factor-alpha (TNF-alpha) neutralizing monoclonal antibody (named as Z12) with TNF-alpha, heavy chain CDR2 (HCDR2) and light chain CDR3 (LCDR3) of Z12 were found to be the most responsible to bind with TNF-alpha. A mimetic peptide (PT) was designed based on the sequence derived from HCDR2 and LCDR3. Fusion protein PT-Fc was constructed by linking PT with Fc of human IgG1 through a flexible linker (GGGGGS). The primary structural characteristics of Fc and PT-Fc were analyzed, including the flexibility, hydrophilicity and epitopes. It was demonstrated that PT and Fc in the fusion protein possessed bio-function properly and non-interfering with each other. Furthermore, PT-Fc was expressed in Escherichia coli by fusion with thioredoxin (Trx). After trx-PT-Fc was cleaved with recombinant enterokinase, PT-Fc was obtained. The results of in vitro cytotoxic assays showed that both PT and PT-Fc could efficiently inhibit TNF-alpha induced apoptosis on L929 cells. At the same micromole concentration, the inhibition activity of PT-Fc was significantly higher than PT. PMID- 15878202 TI - An influx of macrophages is the predominant local immune response in ovine pulmonary adenocarcinoma. AB - Infection with a retrovirus, Jaagsiekte sheep retrovirus (JSRV), causes ovine pulmonary adenocarcinoma (OPA). The excess production of surfactant proteins by alveolar tumour cells results in increased production of pulmonary fluid, which is characteristically expelled through the nostrils of affected sheep. The immune response to JSRV and the tumour is poorly understood: no JSRV-specific circulating antibodies or T cells have been detected to date. The aim of the present study was to obtain phenotypic evidence for a local immune response in OPA lungs. Specific-pathogen free lambs were infected intratracheally with JSRV. When clinical signs of OPA were apparent, the lungs were removed at necropsy and immunohistochemistry (IHC) was performed on lung sections using a panel of mouse anti-sheep mAbs. No influx of dendritic cells, B cells, CD4, CD8 or gammadelta T cells was seen in the neoplastic nodules or in their periphery. MHC Class II positive cells were found intratumourally, peritumourally and in the surrounding alveolar lumina. In the tumours, many of these cells were shown to be fibroblasts and the remainder were likely to be mature macrophages. In the alveolar lumen, the MHC Class II-positive cells were CD14-positive and expressed high levels of IFN-gamma. They appeared to be immature monocytes or macrophages which then differentiated to become CD14-negative as they reached the periphery of the tumours. A high level of MHC Class I expression was detected on a range of cells in the OPA lungs but the tumour nodules themselves contained no MHC Class I positive cells. On the basis of these findings, it is proposed that the lack of an effective immune response in OPA could result from a mechanism of peripheral tolerance in which the activity of the invading macrophages is suppressed by the local environment, possibly as a consequence of the inhibitory properties of the surfactant proteins. PMID- 15878203 TI - An essential role for CCL3 in the development of collagen antibody-induced arthritis. AB - CCL3 is a C-C family chemokine detected at high levels in the synovial tissue and fluids of rheumatoid arthritis (RA) patients. CCL3 binds to the chemokine receptors CCR1 and CCR5, which are expressed by inflammatory leukocytes such as macrophages and T cells present in the affected joints of RA patients. The present study was undertaken to investigate whether absence of CCL3 prevented development of inflammation and joint destruction in anti-type II collagen monoclonal antibody (anti-CII mAb)-induced arthritis. "CCL3 null mice were different from wild-type control mice in terms of body weight loss". In addition, CCL3 null mice exhibited milder clinical and histopathological scores following administration of anti-CII mAb and endotoxin. Moreover, the release of TNF in response to systemic administration of endotoxin was not affected in CCL3 null mice compared to wild-type mice, indicating that the phenotype was not attributable to a defect in endotoxin response. These results indicate that CCL3 plays an essential role in the development of inflammation and joint destruction induced by anti-CII mAb. PMID- 15878204 TI - GABAA receptor beta3 subunit gene-deficient heterozygous mice show parent-of origin and gender-related differences in beta3 subunit levels, EEG, and behavior. AB - The homozygous knockout mouse for the beta3 subunit of the GABAA receptor has been proposed as a model for the neurodevelopmental disorder, Angelman syndrome, based on phenotypic similarities of craniofacial abnormalities, cognitive defects, hyperactivity, motor incoordination, disturbed rest-activity cycles, and epilepsy. Since most children with Angelman syndrome are autosomal heterozygotes of maternal origin, apparently through genomic imprinting, we used gabrb3 deficient heterozygote mice of defined parental origin to investigate whether this phenotype is also maternally imprinted in mouse. Whole brain extracts showed greatly reduced beta3 subunit levels in male mice of maternal origin but not in male mice of paternal origin. Females of both parental origin showed greatly reduced beta3 subunit levels. Heterozygotes did not exhibit hyperactive circling behavior, convulsions, or electrographically recorded seizures. EEGs showed qualitative differences among heterozygotes, with male mice of maternal origin demonstrating more abnormalities including increased theta activity. Ethosuximide inhibited theta bursts, suggesting an alteration in the thalamocortical relay. Carbamazepine induced EEG slowing in males and EEG acceleration in females, with a larger effect in paternal-origin heterozygotes. Evidence thus suggests both parent-of-origin and gender-related components in developmental regulation of beta3 expression, in particular, that the maternally-derived male heterozygote may carry a developmental modification resulting in less beta3 protein, which may reflect partial genomic imprinting of the gabrb3 gene in mice. PMID- 15878205 TI - Predictive value of laryngeal pseudosulcus for gastroesophageal reflux in pediatric patients. AB - OBJECTIVE: To determine if the presence of laryngeal pseudosulcus is a good predictor of reflux in children. METHODS: A retrospective review of clinical photographs of 66 children undergoing direct laryngoscopy for airway symptoms for the presence of laryngeal pseudosulcus. Its presence was correlated with positive reflux tests: 24 h pH monitoring, gastric scintiscan, barium swallow, and esophageal biopsy. RESULTS: Fifty-six patients had a positive reflux test, and 89% of these had pseudosulcus. Seventy percent of the reflux-negative group (N=10) had no pseudosulcus. Sensitivity and specificity were 89% and 70%, respectively. CONCLUSIONS: On its own the presence of pseudosulcus has marginal predictive use, but it is found more frequently among reflux-positive children and may be a useful marker to add to existing visual scoring systems. PMID- 15878206 TI - Connexin 26 mutation 35delG: prevalence of carriers in various regions in France. AB - OBJECTIVE: Mutation 35delG in the connexin 26 gene is the main cause of recessive deafness in Europe. The prevalence of carriers varies, with a mean value proportion of 1/30 in Mediterranean countries. The aim of this study is to determinate the percentage of carriers in several regions of the Mediterranean coast in France. METHODS: This study has been carried out on the genomic DNAs out of a total of 1584 healthy subjects, originating from five French towns or regions, genotyped by Taqman assays. RESULTS: the approximate carrier proportions of the 35delG mutation are 1/50 in Perpignan, 1/65 in Montpellier, 1/66 in Toulon, 1/53 in Grasse. This carrier proportion is 1/31 for the region of Marseilles, a value near the maximal value already obtained in France for Corsica. PMID- 15878207 TI - The effects of sex and neonatal maternal separation on fear-potentiated and light enhanced startle. AB - This study was based on the higher prevalence of anxiety disorders in women than in men, and on the finding that early adverse experiences are a major risk factor for the development of anxiety disorders later in life. The object of this study was to investigate in rats, the sensitivities of the light-enhanced startle (LES) and fear-potentiated startle (FPS) paradigms to sex differences and to determine the effects of maternal separation (MS) on the baseline startle magnitude and potentiated startle response in these paradigms. Pups in the MS group were separated daily from their mother for 180 min/day from postnatal day 2 (PND2) to PND14. Control litters remained undisturbed. The adult male and female progeny were tested in the FPS and LES. As predicted, females showed a significantly greater potentiation of startle than males in the FPS, and a strong trend towards greater startle potentiation in the LES. Contrary to predictions, MS had no effect on startle potentiation in the FPS and severely disrupted LES in female, but not male rats. The observed sex differences add to the validity of the FPS and LES as animal paradigms of fear and anxiety. The findings indicate that these paradigms can be used to study the biological basis of sex differences in fear and anxiety. In contrast, the effects of MS on startle potentiation argue against the idea that MS provides a robust model for the predicted influences of early adverse effects on these startle potentiation measures of fear and anxiety. PMID- 15878208 TI - An essential, putative MEK kinase of Leishmania major. PMID- 15878209 TI - Zonation of the metabolic action of vasopressin in the bivascularly perfused rat liver. AB - Predominance of the vasopressin binding capacity in the hepatic perivenous area leads to the hypothesis that the metabolic effects of the hormone should also be more pronounced in this area. Until now this question has been approached solely by experiments with isolated hepatocytes where an apparent absence of metabolic zonation was found. We have reexamined this question using the bivascularly perfused liver. In this system periportal cells can be reached in a selective manner with substrates and effectors via the hepatic artery when retrograde perfusion (hepatic vein --> portal vein) is done. The action of vasopressin (1-10 nM) on glycogenolysis, initial calcium efflux, glycolysis and oxygen uptake were measured. The results revealed that the action of vasopressin in the liver is heterogeneously distributed. Glycogenolysis stimulation and initial calcium efflux were predominant in the perivenous area, irrespective of the vasopressin concentration. Oxygen uptake was stimulated in the perivenous area; in the periportal area it ranged from inhibition at low vasopressin concentrations to stimulation at high ones. Lactate production was generally greater in the perivenous zone, whereas the opposite occurred with pyruvate production. Analysis of these and other results suggests that at least three factors are contributing to the heterogenic response of the liver parenchyma to vasopressin: a) receptor density, which tends to favour the perivenous zone; b) cell-to-cell interactions, which tend to favour situations where the perivenous zone is amply supplied with vasopressin; and c) the different response capacities of perivenous and periportal cells. PMID- 15878210 TI - Golgi reassembly after mitosis: the AAA family meets the ubiquitin family. AB - The Golgi apparatus in animal cells breaks down at the onset of mitosis and is later rebuilt in the two daughter cells. Two AAA ATPases, NSF and p97/VCP, have been implicated in regulating membrane fusion steps that lead to regrowth of Golgi cisternae from mitotic fragments. NSF dissociates complexes of SNARE proteins, thereby reactivating them to mediate membrane fusion. However, NSF has a second function in regulating SNARE pairing together with the ubiquitin-like protein GATE-16. p97/VCP, on the other hand, is involved in a cycle of ubiquitination and deubiquitination of an unknown target that governs Golgi membrane dynamics. Here, these findings are reviewed and discussed in the context of the increasingly evident role of ubiquitin in membrane traffic processes. PMID- 15878211 TI - Ocular dominancy in conjugate eye movements at reading distance. AB - We recorded conjugate eye movements to elucidate whether ocular dominancy was present at reading distance in 21 normal volunteers with the right-handedness by using a video-oculographic (VOG) measurement. This included the velocity of smooth pursuits, and the latency and velocity of saccades. We defined the dominant eye for each subject by means of the near-far alignment test and 20 subjects showed the right dominant eyes. Although the ocular dominancy was not found in the velocity of smooth pursuit and vertical saccades, the velocity of horizontal saccades in the dominant eyes was faster than that in the non-dominant eyes. These results suggest that the dominant eye is functionally activated prior to non-dominant eye in horizontal saccades at reading distance, which thus indicates the functional dominancy of the dominant eye in conjugate eye movements. PMID- 15878212 TI - Effect of calcium, phosphate and nitrogen on cell growth and biosynthesis of cell wall polysaccharides by Silene vulgaris cell culture. AB - Medium nutrients such as calcium, phosphorus, nitrogen and nitrate to ammonium ratio have significant influence on the growth, biosynthetic and biochemical characteristics of polysaccharides produced by Silene vulgaris (M.) G. cell culture. Cell growth and production of polysaccharides was limited by an absence of any of these components in the medium. Optimal growth of the callus and production of arabinogalactan were achieved at 1.5-4.5 microM calcium while the optimal production of pectin named silenan was observed at 3.0-4.5 microM. The phosphate contents in the medium in the range of 0.63-3.75 microM were favorable for callus growth. Production of silenan was maximal at 1.25-3.75 microM phosphate. Optimal growth of the callus was achieved at 30-90 microM nitrogen. Maximal production of silenan was observed at 60 microM nitrogen while the optimal production of arabinogalactan was at 90 microM nitrogen (at ratio of NH(4)(+):NO(3)(-) as 1:2). A presence both of nitrate and ammonium is needed for the silenan biosynthesis (the NH(4)(+):NO(3)(-) ratio as 1:1 and 1:2). Yields and volumetric production of arabinogalactan were maximal at deletion of ammonium from the nutrient medium (ratio 0:1). Absence of calcium or nitrogen in the medium leads to a decrease of the galacturonic acid residues in silenan. The galactose residues contents in arabinogalactan were decreased in the absence of nitrogen and calcium in the medium. PMID- 15878213 TI - Origin and evolution of viruses causing classical swine fever in Cuba. AB - We have analyzed the origin and evolution of viruses from the classical swine fever (CSF) epidemic that affects Cuba since 2001 by nucleotide sequencing of regions within the E2 glycoprotein and the NS5B (polymerase) genes. The sequence of 190 nucleotides from E2 gene was determined for 10 CSF viruses isolated at different locations of the island, and used for phylogenetic analyses, including sequences from viruses of the 1993--1997 epizootic, previously determined, as well as those from representatives of the different CSFV genotypes. The phylogenetic tree obtained indicates that viruses circulating at present belong to the subgroup 1.2 and are closely related to those isolated during the 1993- 1997 epizootic, including the strain Margarita used for vaccine potency tests in Cuba. However, the pattern of evolution revealed by these analyses was different than that observed previously, in which western isolates were almost identical to Margarita strain, while eastern isolates showed a higher level of genetic diversification. In this case, all the viruses analyzed grouped in an independent, define cluster that is closely related, albeit distinguishable, from that of Margarita-related viruses that previously circulated in the western part of Cuba. In addition, the 2001--2003 viruses showed a branched pattern with a level of sequence diversification similar to that observed in the eastern 1993- 1997 viruses. Interestingly, a significant fraction (about 54%) of the mutations found in the E2 sequence led to amino acid replacements. This high rate of non synonymous mutations was not found in the previous Cuban epizootic and has not been reported for other CSF outbreaks. In spite of these amino acid replacements, no antigenic changes were observed in the reactivity of different isolates with CSFV-specific MAbs and polyclonal sera. The phylogenetic tree derived from 409 nucleotides of NS5B gene of seven isolates and Margarita strain, was consistent with that obtained from E2 sequences. In this region, encoding a non-structural protein, a low level of fixation of non-synonymous mutations was observed. The results obtained suggests that epidemiological factors affecting CSFV spread during the current epizootic in Cuba can favour the fixation of non-synonymous mutation in the E2 gene, which could be associated with a lower severity in the clinical signs developed by most of the affected animals. PMID- 15878214 TI - New features in the genus Ilarvirus revealed by the nucleotide sequence of Fragaria chiloensis latent virus. AB - Fragaria chiloensis latent virus (FClLV), a member of the genus Ilarvirus was first identified in the early 1990s. Double-stranded RNA was extracted from FClLV infected plants and cloned. The complete nucleotide sequence of the virus has been elucidated. RNA 1 encodes a protein with methyltransferase and helicase enzymatic motifs while RNA 2 encodes the viral RNA dependent RNA polymerase and an ORF, that shares no homology with other Ilarvirus genes. RNA 3 codes for movement and coat proteins and an additional ORF, making FClLV possibly the first Ilarvirus encoding a third protein in RNA 3. Phylogenetic analysis reveals that FClLV is most closely related to Prune dwarf virus, the type member of subgroup 4 of the Ilarvirus genus. FClLV is also closely related to Alfalfa mosaic virus (AlMV), a virus that shares many properties with Ilarviruses . We propose the reclassification of AlMV as a member of the Ilarvirus genus instead of being a member of a distinct genus. PMID- 15878215 TI - Hand grip strength in patients with type 2 diabetes mellitus. AB - AIM: The aim of the present study was to compare hand grip strength and pinch power, which are important parameters of hand function, in 76 patients with type 2 diabetes mellitus (T2DM) (mean age: 50.11+/-7.6) with 47 non-diabetic control subjects (mean age: 46.93+/-10.2). METHODS: Grip strength was assessed with a Jamar dynamometer and pinch power was measured with a pinch gauge. Body composition was measured using a Tanita body composition analyzer. Mann-Whitney test, chi-square test, Fisher's exact test, T-test, Kruskal-Wallis variance analysis, Wilcoxon's signed rank test and Pearson's correlation coefficients were used to determine the differences and relations between groups. A p-value <0.05 was taken as statistically significant. RESULTS: Hand grip strength test values were significantly lower in the diabetic group compared with the control group. Key pinch power value for the right hand was significantly lower in the diabetic group than in the control group whereas the left hand value was similar. CONCLUSION: Hand grip strength and key pinch power values were found to be lower in patients with T2DM than in age-matched control subjects. Hands, as well as feet, are also affected by diabetes and physicians should be aware of this. PMID- 15878216 TI - Albumin dialysis reduces portal pressure acutely in patients with severe alcoholic hepatitis. AB - BACKGROUND/AIMS: In patients with alcoholic hepatitis (AH), inflammation contributes to the severity of portal hypertension. This study evaluates the acute effects of albumin dialysis, using the Molecular Adsorbents Recirculating System (MARS), on portal pressure in AH. METHODS: Eleven patients with AH and portal hypertension were treated with MARS (n=8) or haemofiltration (n=3). All patients had associated organ failure manifested by hepatic encephalopathy (Grade 2 or more) or renal failure. Hepatic venous pressure gradient (HVPG) was measured before, during and after the treatment session. RESULTS: A rapid significant reduction of HVPG was observed by 6 h (falling by > or =20% in 7/8 patients, reaching 12 mmHg in 6/8), which was sustained up to 18 h after stopping dialysis. Similar rapid sustained improvements of systemic haemodynamics were also observed. No changes occurred in three patients receiving haemofiltration alone. CONCLUSIONS: Albumin dialysis produces clinically significant, acute reduction in portal pressure but the mechanism by which this effect is achieved is not clear. Our results suggest that MARS may be a useful adjunct in management of portal hypertension, particularly in patients with severe AH with associated organ failure. PMID- 15878217 TI - Effects of 5 daily injections of the neurotensin-mimetic NT69L on the expression of neurotensin receptors in rat brain. AB - The effects of one or five daily intraperitoneal injections of a neurotensin (NT) receptor agonist NT69L (2 mg/kg, i.p.) on the expression of NT (NTS), dopamine 1 and 2 receptors, tyrosine hydroxylase, and DOPA decarboxylase using immunohistochemical and real-time PCR were investigated in rats. Except for the striatum, acute injection of NT69L did not affect neurotensin receptors as compared to saline control. However, 5 daily injections of NT69L resulted in down regulation of both NTS-1 protein and mRNA levels in several brain regions with the striatum showing a dramatic decrease in NTS-1 expression (P<0.05). The down regulation of NTS-1 in the striatum, hypothalamus, and substania nigra (SN) after 5 daily injections was confirmed by autoradiography. Acute injection of NT69L increased NTS-2 mRNA and protein level in prefrontal cortex (PFC). NTS-3 mRNA expression and protein levels were slightly down-regulated in hypothalamus, periaqueductal gray (PAG), and SN, though the difference was not significant. The results indicated a difference in the profile of NT receptors expression in response to NT69L. Tyrosine hydroxylase (TH) and DOPA decarboxylase (DDC) mRNA was significantly down-regulated in striatum but not in SN. Interestingly, Nurr 1, a transcriptional activator of TH, was dramatically up-regulated in striatum, but down-regulated in PFC, suggesting that different modulating mechanisms may participate in NT69L tolerance in different regions. The present results suggest that distinct NT receptors involved in the effects exerted by NT69L may contribute to the interactions of NT69L with both neural networks and cellular proteins. PMID- 15878218 TI - Synthesis and antimalarial activity of E-2-quinolinylbenzocycloalcanones. AB - A series of E-2-quinolinylbenzocycloalcanones 5-21 were prepared and evaluated for their activity to inhibit beta-hematin formation and the hydrolysis of hemoglobin in vitro. Positive compounds for both assays were also tested for their efficacy in rodent Plasmodium berghei. Compounds 6, 16, 19, and 20, were the most promising. Inhibition of beta-hematin formation was minimal when a hydrogen or methoxy groups were present on the position 8 of the quinoline and position 4' of the indanone ring as it appeared for compounds 5, 7-15, 17, 18, and 21, and greatest with compounds (52%) and (90%) with a substitution of methoxy on position 6 and 7 or methyl on position 8 of the quinoline nucleus and methoxy or methyl groups on position 4' of the indanone. The most active compound to emerge from this study is 2-chloro-8-methyl-3-[(4'-methoxy-1'-indanoyl)-2' methyliden]-quinoline 20 effective as antimalarial that target beta-hematin formation and the inhibition of the hydrolysis of hemoglobin in vitro together with a good survival in a murine malaria model, which should help delay the rapid onset of resistance to drugs acting at only a single site. Results with these assays suggest that quinolinylbenzocycloalcanones exert their antimalarial activity via multiple mechanisms. PMID- 15878219 TI - 2-Arylidene-4-(4-phenoxy-phenyl)but-3-en-4-olides: synthesis, reactions and biological activity. AB - 2-Arylidene-4-(4-phenoxy-phenyl)but-3-en-4-olides (1-17) were prepared from 3-(4 phenoxy-benzoyl)propionic acid and aromatic aldehydes. Some of the selected butenolides were reacted with ammonia and benzylamine to give corresponding 3 arylidene-5-(4-phenoxy-phenyl)-2(3H)-pyrrolones (18-23) and 3-arylidene-5-(4 phenoxy-phenyl)-1-benzyl-2(3H)-pyrrolones (24-29) respectively, which were characterized on the basis of 1H-, 13C-NMR, Mass spectrometric data and elemental analysis results. These compounds were tested for anti-inflammatory and antimicrobial actions. The compounds, which showed significant anti-inflammatory activity, were screened for their analgesic and ulcerogenic activities. Five new compounds (5, 6, 7, 25 and 26), out of 29 showed very good anti-inflammatory activity in the carrageenan induced rat paw edema test, with significant analgesic activity in the acetic acid induced writhing test together with negligible ulcerogenic action. Antibacterial activity against Staphylococcus aureus and Escherichia coli as well as antifungal activity against Candida albicans were expressed as the corresponding minimum inhibitory concentration (MIC) values. Compound 21, 22 and 23 showed excellent activity against C. albicans with MIC-10 microg/ml. Out of the above-mentioned compounds, 22 and 23 also showed good activity against S. aureus with MIC-20 and 15 microg/ml respectively. PMID- 15878220 TI - Synthesis, spectroscopic and biological properties of bis(3-arylimidazolidinyl 1)methanes. A novel family of antimicrobial agents. AB - Synthesis, spectroscopic and biological properties of new bis(3 arylimidazolidinyl-1)methanes are described. These compounds were synthesized by condensation reaction between N-arylethylenediamines and formaldehyde. Chemical structures were confirmed by means of their (1)H- and (13)C-NMR and mass spectroscopic data. Investigation of in vitro antimicrobial activity was performed using Gram-negative and Gram-positive bacteria as well as antifungal studies against Aspergillus niger and Candida albicans. Minimal inhibitory concentrations of active compounds were determined. PMID- 15878221 TI - [Ethics of medical decisions]. PMID- 15878222 TI - [An unknown cause of elevated chromogranin A levels: the proton-pump inhibitors]. PMID- 15878223 TI - [Internists reunited on the Loire Board]. PMID- 15878224 TI - A quasi-dynamic nonlinear finite element model to investigate prosthetic interface stresses during walking for trans-tibial amputees. AB - BACKGROUND: To predict the interface pressure between residual limb and prosthetic socket for trans-tibial amputees during walking. METHODS: A quasi dynamic finite element model was built based on the actual geometry of residual limb, internal bones and socket liner. To simulate the friction/slip boundary conditions between the skin and liner, automated surface-to-surface contact was used. Besides variable external loads and material inertia, the coupling between the large rigid displacement of knee joint and small elastic deformation of residual limb and prosthetic components were also considered. RESULTS: Interface pressure distribution was found to have the same profile during walking. The high pressures fall over popliteal depression, middle patella tendon, lateral tibia and medial tibia regions. Interface pressure predicted by static or quasi-dynamic analysis had the similar double-peaked waveform shape in stance phase. INTERPRETATION: The consideration of inertial effects and motion of knee joint cause 210% average variation of the area between the pressure curve and the horizontal line of pressure threshold between two cases, even though there is only a small change in the peak pressure. The findings in this paper show that the coupling dynamic effects of inertial loads and knee flexion must be considered to study interface pressure between residual limb and prosthetic socket during walking. PMID- 15878225 TI - Toxicologic evaluation of licorice extract as a cigarette ingredient. AB - Licorice extract (block, powder or liquid) may be applied to cigarette tobacco at levels of about 1-4% to enhance and harmonize the flavor characteristics of smoke, improve moisture holding characteristics of tobacco, and act as a surface active agent for ingredient application. Neat material pyrolysis studies, and smoke chemistry and biological activity studies (bacterial mutagenicity, cytotoxicity, micronucleus, and sub-chronic inhalation) with mainstream smoke, or mainstream smoke preparations from cigarettes containing various target levels (1.5-12%) of the licorice extracts were performed to provide data for an assessment of the use of licorice extract as a cigarette tobacco ingredient. At simulated tobacco burning temperatures up to 900 degrees C all forms of neat licorice extract pyrolyzed extensively, yielding small amounts of benzene, toluene, phenol and acetaldehyde with no indication that licorice extracts would transfer intact to mainstream smoke. As a single ingredient added to cigarette tobacco, block licorice extract at a target level of 12.5% increased smoke constituents including selected PAH, arsenic, lead, phenol and formaldehyde (on a TPM basis), while licorice extract powder (target level of 8% tobacco) increased select PAH, phenol and formaldehyde (on a TPM basis). Lower target application levels (including typical application levels) of block, powder or liquid licorice extract did not significantly alter the smoke chemistry profile. Biological tests indicated no relevant difference in the genotoxic or cytotoxic potential of either mainstream smoke (or smoke preparations) from cigarettes with added licorice extracts compared to control cigarettes. In sub-chronic 90-day rat inhalation studies, the mainstream smoke from cigarettes with 12.5% added block and 8% added powder licorice extract contained higher formaldehyde concentrations compared to control cigarette smoke. Female rats in the 12.5% block licorice extract exposure group displayed an increased incidence and severity of epithelial hyperplasia in the nose (level 2), with no relevant respiratory tract changes in the 8% powder licorice extract exposed rats. At the lower licorice extract application levels (1.25-5%), there was no indication of increased formaldehyde concentration in the smoke atmosphere and no relevant changes in respiratory tract tissues. Mineralcorticoid-like effects which have been associated with excess licorice ingestion were not found in any of the smoke inhalation studies. The results of these studies with various forms of licorice extract applied to cigarette tobacco suggest that adding licorice extract to cigarette tobacco at levels of < or =5% does not discernibly alter the smoke chemistry or biological effects normally associated with mainstream cigarette smoke. PMID- 15878226 TI - [Our experience about ten end-to-side sensitive nerve sutures at the hand]. AB - Ten traumatic nerve defects at the palm or digit level were treated by end-to side (ETS) neurorrhaphy. The technique included removal of an epineurial window on the donor nerve and coaptation of the sharply cut distal end of the injured nerve by epineurial stitches under microscope magnification. At final follow-up, the static two-point discrimination test scored at an average of 9.1 (range, 6 12) mm on the repaired nerve, compared to an average of 4.6 (range, 4-6) mm on the contralateral uninjured control side. The moving two-point discrimination test scored at an average of 7 (range, 4-10) mm on the repaired nerve compared to an average of 2.6 (range, 2-4) mm on the control side. This short series showed ETS neurorrhaphies are effective and give comparable results with those of nerve grafts or vein conduits, but worse than direct repairs. PMID- 15878227 TI - [Mondor's disease: about two cases]. AB - The authors report two cases of Mondor's disease after mammoplasty reduction's surgery. This pathology rare and benign should be known by all surgeons, because it could be a postoperative discord, especially in these lithigious times. If the evolution is favourable in one or two months, the therapeutic urgency is to reassure the patient. PMID- 15878228 TI - An European inter-laboratory validation of alternative endpoints of the murine local lymph node assay: 2nd round. AB - The original local lymph node assay (LLNA) is based on the use of radioactive labelling to measure cell proliferation. Other endpoints for the assessment of proliferation are also authorized by the OECD Guideline 429 provided there is appropriate scientific support, including full citations and description of the methodology (OECD, 2002. OECD Guideline for the Testing of Chemicals; Skin Sensitization: Local Lymph Node Assay, Guideline 429. Paris, adopted 24th April 2002.). Here, we describe the outcome of the second round of an inter-laboratory validation of alternative endpoints in the LLNA conducted in nine laboratories in Europe. The validation study was managed and supervised by the Swiss Agency for Therapeutic Products (Swissmedic) in Bern. Ear-draining lymph node (LN) weight and cell counts were used to assess LN cell proliferation instead of [3H]TdR incorporation. In addition, the acute inflammatory skin reaction was measured by ear weight determination of circular biopsies of the ears to identify skin irritation properties of the test items. The statistical analysis was performed in the department of statistics at the university of Bern. Similar to the EC(3) values defined for the radioactive method, threshold values were calculated for the endpoints measured in this modification of the LLNA. It was concluded that all parameters measured have to be taken into consideration for the categorisation of compounds due to their sensitising potencies. Therefore, an assessment scheme has been developed which turned out to be of great importance to consistently assess sensitisation versus irritancy based on the data of the different parameters. In contrast to the radioactive method, irritants have been picked up by all the laboratories applying this assessment scheme. PMID- 15878229 TI - Sea bass (Dicentrarchus labrax) androgen receptor: cDNA cloning, tissue-specific expression, and mRNA levels during early development and sex differentiation. AB - Androgens play key roles in vertebrate sex differentiation, gonadal maturation and reproductive behaviour and their actions are generally mediated through specific nuclear receptors. The present study describes the isolation, sequencing and characterization of the cDNA encoding the androgen receptor (AR) in the European sea bass. AR was cloned from a sea bass testis cDNA library and encoded a predicted protein of 767 residues, with a calculated molecular weight of 86.4 kDa and a theoretical pI of 6.34. Several domains present in all cloned ARs were identified. The domains corresponded to an amino-terminal hypervariable transcriptional activation domain (TAD), a central highly conserved DNA-binding domain (DBD), and a carboxy-terminal ligand-binding domain (LBD). Percentages of homology-similarity among these functional domains in teleost fish ranged between 9 and 75% for the TAD, 73 and 98% for the DBD, and 78 and 96% for the LBD when compared to those of the sea bass. Tissue-specific expression showed that AR was preferentially expressed in testis, ovaries, and brain. Some other tissues such as the head kidney, liver and spleen also showed AR expression although at very low levels. A semiquantitative PCR was developed to study the expression of AR mRNA during the period of development encompassed between 50 and 300 DPH in sea bass gonads. An experimental design, involving repeated size gradings, based on the fact that sea bass females are larger than males already at sex differentiation, was set to obtain a group consisting of the largest fish (female dominant) and a group consisting of the smallest fish (male-dominant). The results showed very low mRNA expression levels of AR in the gonads during early development. Differences in AR expression between groups were first encountered at 150 DPH and became especially marked at 250 DPH with much higher levels in the male-dominant group. These sex-related differences in expression profiles between males and females by the time of sex differentiation, suggest an important role for AR controlling this process in the sea bass. PMID- 15878230 TI - Thyroid dysfunction in megalin deficient mice. AB - Megalin mediates transcytosis of thyroglobulin (Tg), the thyroid hormone precursor, resulting in its passage into the bloodstream. The process involves especially hormone-poor Tg, which may favour hormone secretion by preventing competition with hormone-rich Tg for proteolytic degradation. To gain more insight into the role of megalin, here we studied thyroid function and histology in megalin deficient mice compared with WT mice. As expected from the knowledge that megalin mediates Tg transcytosis, serum Tg levels were significantly reduced in homozygous (megalin-/-) mice, which, more importantly, were found to be hypothyroid, as demonstrated by significantly reduced serum free thyroxine and significantly increased serum thyroid stimulating hormone (TSH) levels. In heterozygous (megalin+/-) mice, in which megalin expression was normal, thyroid function was unaffected. Although the serological phenotype in megalin-/- mice was not associated with histological alterations or goiter, our results support a major role of megalin in thyroid hormone secretion. PMID- 15878231 TI - Midkine, a heparin-binding growth factor, produced by the host enhances metastasis of Lewis lung carcinoma cells. AB - Midkine is a heparin-binding growth factor and is expressed by a number of tumor cells, contributing to their growth both in vitro and in vivo. Spontaneous lung metastasis of Lewis lung carcinoma cells, which did not significantly express MK, was significantly less extensive in mice deficient in the midkine gene than in wild-type mice, when the tumor was subcutaneously grown above the thigh. Midkine strongly enhanced migration of Lewis lung carcinoma cells in vitro. Therefore, midkine is also a host factor enhancing tumor metastasis. Anti-midkine therapy for malignancy may act on midkine produced by both the tumor and host. PMID- 15878232 TI - A truncated splice variant of human BARD1 that lacks the RING finger and ankyrin repeats. AB - BARD1 is a crucial partner of the breast and ovarian tumor suppressor BRCA1 required for ubiquitin ligase activity and for reciprocal stabilization in cells. We report here an alternatively spliced human BARD1 mRNA variant (BARD1DeltaRIN) isolated from a HeLa cell cDNA library. It is characterized by deletion of exons 2-6 that encode most of the RING finger domain and the entire span of ankyrin repeats. DeltaRIN transcript was detected in all breast cancer-cell lines studied although its protein expression level was low. DeltaRIN does not interact with BRCA1, whereas it interacts with and colocalizes with CstF-50 to cytoplasmic dots. Hence, a deletion variant of BARD1 occurs in cells and may play a distinct role with CstF-50. PMID- 15878233 TI - Use of advanced oxidation processes to improve the biodegradability of mature landfill leachates. AB - Two advanced oxidative processes (Fe2+/H2O2/UV and H2O2/UV systems) were used for the pre-treatment of mature landfill leachate with the objective of improving its overall biodegradability, evaluated in terms of BOD5/COD ratio, up to a value compatible with biological treatment. At optimized experimental conditions (2000 mgL(-1) of H2O2 and 10 mgL(-1) of Fe2+ for the photo-Fenton system, and 3000 mgL( 1) of H2O2 for the H2O2/UV system), both methods showed suitability for partial removal of chemical oxygen demand (COD), total organic carbon (TOC) and color. The biodegradability was significantly improved (BOD5/COD from 0.13 to 0.37 or 0.42) which allowed an almost total removal of COD and color by a sequential activated sludge process. In addition, gel permeation chromatography (GPC) has showed a substantial agreement on the cleavage of large organic compound into smaller ones. PMID- 15878234 TI - Adsorption of copper from aqueous solution on Brassica cumpestris (mustard oil cake). AB - The adsorption behavior of various heavy metals on mustard oil cake (MOC) was studied. The maximum adsorption of Cu(II) was observed followed by Zn(II), Cr(VI), Mn(II), Cd(II), Ni(II) and Pb(II). The adsorption of Cu(II) was found to be dependent on initial concentration of solution, pH, adsorbent dose, temperature and contact time. The adsorption followed pseudo-first-order and second-order kinetics but pseudo-second-order kinetic model was better obeyed since experimental data agreed well with theoretical data. Thermodynamic parameters were also evaluated. The adsorption process was found to be endothermic and spontaneous in nature. Attempts were also made to desorb Cu(II) from the adsorbent and regeneration of the spent adsorbent. The breakthrough and exhaustive capacities were found to be 5 and 10 mg g(-1), respectively. PMID- 15878235 TI - Dynamic stability of locomotor respiratory coupling during cycling in humans. AB - We explored the locomotor respiratory coupling (LRC) during a 50-min constant load submaximal cycling exercise. A 4-week recombinant human erythropoietin (r HuEPO) treatment improved participants' aerobic capabilities, but did not elicit significant changes in LRC. The distributions of the respiratory frequency over pedalling frequency ratios were systematically bimodal, with a preferred use of 1/3 and 1/2, and a progressive shift of the higher mode from 1/3 towards 1/2 with exercise duration. These results are interpreted in the framework of the sine circle map as the result of coordination dynamics between the physiological subsystems involved in the breathing pedalling cooperation. PMID- 15878236 TI - Olfactory stimulation with scent of lavender oil affects autonomic nerves, lipolysis and appetite in rats. AB - In a previous study, we presented evidence that scent of grapefruit oil excites sympathetic nerves innervating white and brown adipose tissues and the adrenal gland, inhibits the vagal nerve innervating the stomach, increases lipolysis and heat production (energy consumption), and reduces appetite and body weight. Here, we examined the effects of olfactory stimulation with scent of lavender oil (SLVO) in rats and observed that in contrast to grapefruit oil, it inhibits the sympathetic nerves innervating the white and brown adipose tissues and adrenal gland and excites the parasympathetic gastric nerve. Local anesthesia of the nasal mucosa with xylocaine or anosmic treatment using ZnSO(4) eliminated the autonomic changes caused by SLVO. Moreover, stimulation with SLVO lowered the plasma glycerol level, and treatment with either ZnSO(4) or an intracranial injection of thioperamide, a histamine H3 receptor-antagonist, abolished SLVO mediated glycerol decline. Furthermore, a 15-min daily exposure to SLVO increased food intake and body weight. Finally, linalool, a component of lavender oil, induced responses similar to those caused by SLVO, and the glycerol response to linalool was eliminated by thioperamide. Thus, scent of lavender oil and its active component, linalool, affect autonomic nerves, suppress lipolysis through a histaminergic response, and enhance appetite and body weight. PMID- 15878237 TI - Adaptation to hydrogen peroxide enhances PC12 cell tolerance against oxidative damage. AB - The adaptive responses to H2O2 and the resulting protective effect against oxidative stress have been investigated using PC12 cells. Pretreatment of sublethal doses of H2O2 significantly protected PC12 cells against the cytotoxicity induced by lethal H2O2. The endogenous antioxidant defense systems, catalase and glutathione peroxidase, were enhanced quickly by the pretreatment of low doses of H2O2. This pretreatment also exerted protective effect against the oxidative insults induced by 6-hydroxydopamine and paraquat, but not against alkyl peroxyl radicals. Our results, taken together, suggest that the stimulation by low dose of H2O2 enriches the cellular antioxidant defense systems, thereby enhancing cell tolerance against the forthcoming oxidative insults induced by H2O2 and related hydroxyl radicals. PMID- 15878238 TI - Imaging of ferroelectric thin films by X-ray photoemission electron microscopy (XPEEM). AB - We performed X-ray photoemission electron microscopy (XPEEM) measurements at the Nanospectroscopy Beamline of the synchrotron light source ELETTRA, Trieste, Italy, to demonstrate the principal possibility of imaging ferroelectric thin films by low-energy photoelectrons. Due to the insulating properties of ferroelectric films, severe surface charging was the major experimental challenge to overcome. This was achieved by grounding an array of gold inter-digital electrodes (with 5 microm blank intervals between them) deposited on top of the films. The images taken with BaTiO(3) films revealed 50-100 nm-sized holes (material discontinuities) on the surface, an observation confirmed by high resolution scanning electron microscopy (HRSEM). Finer details, e.g. a granular structure, which has been resolved with HRSEM, could not be observed in the XPEEM images. Our measurements indicate that despite some residual charging, a 50 nm lateral resolution can be achieved in XPEEM measurements with ferroelectric films. PMID- 15878239 TI - Isolation of Cryptosporidium andersoni Kawatabi type in a slaughterhouse in the northern island of Japan. AB - Fecal samples were collected from 325 adult cattle and 108 pigs in a slaughterhouse in Hokkaido, the northern island of Japan. Five adult cattle were found to be positive for oocysts of Cryptopsoridium (1.5%). The oocysts were morphologically similar to those of Cryptosporidium andersoni. The partial sequence of the 18S rRNA gene of the isolate was 100% identical with that of the C. andersoni Kawatabi strain. SCID mice were infected after oral administration. Based on the morphology of the oocysts, the sequence of the 18S rRNA gene and the infectivity to SCID mice, the isolate was concluded to be of the same type as the C. andersoni Kawatabi strain that has been isolated in Honshu, the main island of Japan. PMID- 15878240 TI - A survey of alternative transcripts of human tissue kallikrein genes. AB - Alternative splicing is prevalent within the human tissue kallikrein gene locus. Aside from being the most important source of protein diversity in eukaryotes, this process plays a significant role in development, physiology and disease. A better understanding of alternative splicing could lead to the use of gene variants as drug targets, therapeutic agents or diagnostic markers. With the rapidly rising number of alternative kallikrein transcripts, classifying new transcripts and piecing together the significance of existing data are becoming increasingly challenging. In this review, we present a systematic analysis of all currently known kallikrein alternative transcripts. By defining a reference form for each of the 15 kallikrein genes (KLK1 to KLK15), we were able to classify alternative splicing patterns. We identified 82 different kallikrein gene transcript forms, including reference forms. Alternative splicing may lead to the synthesis of 56 different protein forms for KLK1-15. In the kallikrein locus, the majority of alternative splicing events occur within the protein-coding region, and to a lesser extent in the 5' untranslated regions (UTRs). The most common alternative splicing event is exon skipping (35%) and the least common events are cryptic exons (3%) and internal exon deletion (3%). Seventy-six percent of kallikrein splice variants that are predicted to encode truncated proteins are the result of frameshifts. Eighty-nine percent of putative proteins encoded by splice variants are predicted to be secreted. Although several reports describe the identification of kallikrein splice variants and their potential clinical utility, this is the first extensive review on this subject. Accumulating evidence suggests that alternative kallikrein forms could be involved in many pathologic conditions or could have practical applications as biomarkers. The organization and analysis of the kallikrein transcripts will facilitate future work in this area and may lead to novel clinical and diagnostic applications. PMID- 15878241 TI - Alterations in dendritic morphology of prefrontal cortical and nucleus accumbens neurons in post-pubertal rats after neonatal excitotoxic lesions of the ventral hippocampus. AB - Neonatal ventral hippocampal (nVH) lesions in rats result in adult onset of a number of behavioral and cognitive abnormalities analogous to those seen in schizophrenia, including hyperresponsiveness to stress and psychostimulants and deficits in working memory, sensorimotor gating and social interaction. Molecular and neurochemical alterations in the prefrontal cortex (PFC) and nucleus accumbens (NAcc) of nVH-lesioned animals suggest developmental reorganization of these structures following neonatal lesions. To determine whether nVH lesions lead to neuronal morphological changes, we investigated the effect of nVH lesion on dendritic structure and spine density of pyramidal neurons of the PFC and medium spiny neurons of the NAcc. Bilateral ibotenic acid-induced lesion of the VH was made in Sprague-Dawley pups at postnatal day 7 (P7); and at P70, neuronal morphology was quantified by modified Golgi-Cox staining. The results show that length of basilar dendrites and branching and the density of dendritic spines on layer 3 pyramidal neurons were significantly decreased in rats with nVH lesions. Medium spiny neurons from the NAcc showed a decrease in the density of dendritic spines without significant changes in dendritic length or arborization. The data, comparable to those observed in the PFC of schizophrenic patients, suggest that developmental loss of excitatory projections from the VH may lead to altered neuronal plasticity in the PFC and the NAcc that may contribute to the behavioral changes in these animals. PMID- 15878242 TI - Co-expression of the P75 neurotrophin receptor and neurotrophin receptor interacting melanoma antigen homolog in the mature rat brain. AB - The p75 neurotrophin receptor (p75(NTR)) is involved in the regulation of neuronal survival and phenotype, but its signal transduction mechanisms are poorly understood. Recent evidence has implicated the cytoplasmic protein NRAGE (neurotrophin receptor-interacting MAGE (from Melanoma AntiGEn) homolog) in p75(NTR) signaling. To gain further insight into the role of NRAGE, we investigated the co-expression of NRAGE and p75(NTR) in mature rat brain. In all areas examined, NRAGE appeared to be confined to neurons. In the basal forebrain cholinergic complex, NRAGE immunoreactivity was evident in all p75(NTR)-positive neurons. There were many more NRAGE-positive than p75(NTR)-positive neurons in these regions, however. NRAGE was also expressed in areas of the basal forebrain that did not express p75(NTR), including the lateral septal nucleus and the nucleus accumbens. A finding in marked contrast to previous studies was the presence of p75(NTR) immunoreactivity in neuronal cell bodies in the hippocampus. Hippocampal p75(NTR) immunoreactivity was apparent in rats 6 months and older, and was localized to the dentate gyrus and stratum oriens. All p75(NTR)-positive neurons in the dentate gyrus and hippocampal formation were positive for NRAGE. The majority of granular cells of the dentate gyrus and pyramidal cells in the hippocampal formation were positive for NRAGE and negative for p75(NTR). NRAGE was also present in some neuronal populations that express p75(NTR) after injury, including striatal cholinergic interneurons, and motor neurons. A region of marked disparity was the cerebral cortex, in which NRAGE immunoreactivity was widespread whereas p75(NTR) was absent. The results are consistent with an important role for NRAGE in p75(NTR) signaling, as all cells that expressed p75(NTR) also expressed NRAGE. The wider distribution of NRAGE expression suggests that NRAGE may also participate in other signaling processes. PMID- 15878243 TI - Pathophysiology of newborn hypotension outside the transitional period. AB - Hypotension is a common diagnosis in neonatal intensive care units. Although there are epidemiological data for normative blood pressure values in preterm and term infants, the ranges of normal blood pressure where adequate organ perfusion is ensured for different gestational and postnatal ages remain unclear. An understanding of the developmentally regulated differences in the physiology and pathophysiology of the neonatal cardiovascular system in comparison to that of mature subjects is important to formulate an appropriate treatment strategy in neonates with circulatory compromise. This article reviews the current understanding of pathophysiology of hypotension and shock in the neonate beyond the transitional period, focusing on hypovolemia, myocardial dysfunction, abnormal peripheral vasoregulation, and relative or absolute adrenal insufficiency with potentially associated down regulation of adrenergic receptors. PMID- 15878244 TI - Cross-linked high amylose starch derivatives for drug release III. Diffusion properties. AB - Acetate (Ac-), aminoethyl (AE-) and carboxymethyl (CM-) derivatives of cross linked high amylose starch (HASCL-6) were previously shown to control the release of drugs over 20 h from highly loaded (up to 60% drug) monolithic tablets. This report presents a diffusion analysis, aimed to facilitate a better understanding of the mechanisms involved in the control of the drug release from these hydrogels. The diffusion was found to depend on the molecular weight of the diffusant, whereas the partition coefficient depended on the affinities of the diffusant for the polymers and for the dissolution media via attractive or repulsive ionic interactions. The diffusion was also affected by the swelling of CM-HASCL-6, which, unexpectedly, increased with the decrease of the ionic strength. This diffusion analysis completes the swelling studies of HASCL-6 and of its derivatives, allowing the prediction of release kinetics of various active agents. PMID- 15878245 TI - Evaluation of plants used for antimalarial treatment by the Maasai of Kenya. AB - Semi-structured interviews with three Maasai herbalists led to the identification and collection of 21 species of plants used to treat malaria. Extracts were evaluated using in vitro antimalarial and cytotoxicity assays. Of the species tested, over half were antiplasmodial (IC50<10 microg/ml), and all but one (Gutenbergia cordifolia Benth.) displayed selectivity for the malaria parasite Plasmodium falciparum as indicated by a lack of cytotoxicity (ED50>20 microg/ml) against cultured KB cells. The results of this preliminary investigation support the traditional knowledge of Maasai herbalists and justify ethnomedical inquiry as a promising method, specifically, in antimalarial therapy, to yield leads for drug discovery. PMID- 15878246 TI - Folk phytotherapeutical plants from Maratea area (Basilicata, Italy). AB - Field ethnobotanical survey was undertaken for the period of 2002-2003 in the Tyrrhenian part of the Basilicata region of southern Italy. Data of 56 species of plants belonging to 29 families where gathered through interviews; among the species, 47 are used in human therapy, 6 as insect repellents, 15 in veterinary medicine, 1 for its ichthyotoxic properties and 3 for magic therapeutic purposes. The most important findings in ethnomedicine relate to Nasturtium officinale (renal colic, liver diseases), Foeniculum vulgare subsp. piperitum (mouth ulcers), Leopoldia comosa (toothache, headache), Micromeria graeca subsp. graeca (coughs) and Ceterach officinarum (malaria), while in the ethnoveterinary field, we have Pteridium aquilinum (wolf bites) and Spartium junceum (fractures of animal limbs). PMID- 15878247 TI - Antibacterial potential from Indian Suregada angustifolia. AB - Phytochemical analysis of the leaves from Indian Suregada angustifolia (Baill. ex Muell. Arg.) Airy Shaw (Euphorbiaceae) resulted in the isolation and identification of six known compounds, viz. friedelin, epi-friedelinol, n octacosanol, alpha-amyrin, beta-sitosterol and beta-sitosterol-3-beta-D glucopyranoside. Aqueous (room temperature, boiled and autoclaved) and various solvent (methanol, chloroform and hexane) extracts of leaves were tested against 12 human pathogenic bacteria by the agar well-diffusion method. Aqueous extracts did not express any activity. Antibacterial activity was recorded in the order of methanol, hexane and chloroform extracts. Maximum activity recorded against Staphylococcus aureus (skin infections) in methanol and hexane extracts and moderate activity recorded against diarrhoea causing bacteria, Vibrio vulnificus (hexane extract) and Vibrio cholerae (chloroform extract). PMID- 15878248 TI - Serial MR imaging and 1H-MR spectroscopy of unidentified bright objects in a case of neurofibromatosis type 1. AB - Serial study using MR imaging and 1H-MR spectroscopy (1H-MRS) of unidentified bright objects (UBO) in a 9-year-old boy with neurofibromatosis type 1 (NF1) is described. UBO was indicated in the left globus pallidum at 3 years old, then appeared in the right globus pallidum at 5 years old along with left hemiconvulsion. These UBO gradually disappeared over several years. On 1H-MRS, the ratios of N-acetylaspartate (NAA)/creatine and that of choline/creatine were each mildly reduced in the UBO. These ratios improved in parallel with disappearance of the UBO. In addition, elevation of the lactate/lipid peak was shown in UBO during the early stage, however, it disappeared in the latter. The longitudinal follow-up using MR image and 1H-MRS was useful for metabolic evaluation of UBO in patients with NF1. PMID- 15878249 TI - [Surgical wound infections due to Pseudomonas aeruginosa in orthopedic surgery]. AB - OBJECTIVE: The department of infection control carried out an investigation to search for the origin of 4 surgical site infections and 1 wound colonization by Pseudomonas aeruginosa in patients having undergone orthopedic surgery. PATIENTS AND METHODS: The authors retrospectively reviewed the medical records, the clinical data of the operating units, as well as the bacteriological assessments of the infected patients. Multiple environmental samples were made to screen for P. aeruginosa and care giving was evaluated. RESULTS: The 5 patients underwent surgery between August and September 2001 with various surgeons and were followed up by various paramedics. The surgical procedures were varied and performed in different operating rooms. Various P. aeruginosa serotypes were isolated. No specific event could be related to the infections concerning the surgical procedures. In 3 of the 5 patients, non-sterile cotton jersey had been used, either normally (plaster or plaster splint) or after sterilization (wrapping of wounded limbs before surgical procedure). The culture samplings of non-sterile jersey were always contaminated by Enterobacteriaceae or Pseudomonas sp., with 2 positives cultures of P. aeruginosa. Only one water sample was positive, whereas other environmental samples remained negative. The reorganization of jersey supply put an end to this epidemic phenomenon. CONCLUSION: The most probable hypothesis for surgical wound infection was the cotton jersey in 3 of the 5 cases. PMID- 15878250 TI - [Adaptive behaviors to HIV risk of transmission in different populations]. AB - Since the beginning of the HIV epidemic in France, surveys aimed at better understanding risk perceptions of HIV infection and preventive sexual behaviors have been implemented in the general population, and in populations such as IVDU and homosexual men, more concerned by risks of HIV transmission. The objective of this article is to describe these surveys, to present their main results and to assess what has been the overall impact of prevention campaigns on the adoption of preventive sexual behaviors in these populations. The results show that very early after the beginning of the AIDS epidemic, both general and homosexual populations have adopted preventive sexual behaviors, mainly increasing condom use and implementing other preventive strategies. However, with the introduction of HAART in 1996, a slackening of these preventive behaviors is noted. The use of condom is less frequent, especially in the youngest generations of both general and homosexual populations. On the opposite, among IVDU, the use of sterile syringes increased dramatically as soon as over-the-counter sales of syringes was authorized in 1987, as well as the adoption of ways other than intravenous to take drugs. Both have contributed to almost stop the HIV epidemic in this specific group. The results of these surveys show that the benefits of prevention campaigns are different between populations and are reversible. It is necessary to renew the messages, campaigns and programs of prevention with the renewal of generations. It is also necessary to adapt these messages to the new scientific data, and to the evolution of social and individual representations of the disease. PMID- 15878251 TI - [The role of primary care physicians in the detection and control of epidemics: the Geneva experience]. AB - Private practice physicians play a central role in the detection and control of outbreaks. In Geneva, a local alert and response network completes the federal surveillance system, based on the mandatory reporting of infectious diseases. The common objective of these two mechanisms is to help health authorities in detecting, assessing, and controlling infectious diseases treats. Private practice physicians are one of the most effective and timely sources of information. As network members, they receive brief reports and information and are involved in the implementation of control and prevention measures. Their key role was confirmed during a measles epidemic which occurred in Geneva in the spring of 2003. PMID- 15878252 TI - Development and optimization of a rapid HPLC method for analysis of ricobendazole and albendazole sulfone in sheep plasma. AB - A simple, rapid and reliable high performance liquid chromatographic (HPLC) method has been developed and validated for simultaneous determination of ricobendazole (RBZ) and its main metabolite albendazole sulfone (ABZSO(2)) in sheep plasma using an isocratic system with UV detection. The method involved solid phase extraction followed by separation on a reversed phase C-18 column. Internal standard was selected by quantitative structure retention relationships (QSRRs) analysis. A method to optimize the composition of ternary components mobile phase with the assistance of multiple linear regression is described. Retention times were within 10 min. The calibration curves were linear over a concentration range of 10-1000 ng/ml for both RBZ and ABZSO(2) (r > 0.999). Intra day relative standard deviation at low, medium and high concentration levels were <5.5% for RBZ and <4.6% for ABZSO(2); average accuracies were 98.3, 101.0 and 100.5% for RBZ and 101.0, 102.4 and 100.8% for ABZSO(2). The inter-day variations at the same concentrations were <5.9% for RBZ and <6.4% for ABZSO(2). The extraction recoveries at these concentrations for RBZ, ABZSO(2) and the internal standard were all over 96%. The limit of detection and limit of quantitation were 2.4 and 7.1 ng/ml, respectively for RBZ, and 10ng/ml for both analytes. PMID- 15878253 TI - Expression and activation of peroxisome proliferator-activated receptors in growth plate chondrocytes. AB - Peroxisome proliferator-activated receptor gamma (PPAR-gamma) is a nuclear hormone receptor that is involved in a wide range of cellular processes. Although it is known that PPAR-gamma plays an important role in cell cycle control, inflammation, apoptotic cell death, and other cellular processes, the role of PPAR-gamma in the normal and pathological function of growth plate chondrocytes has not been investigated. The purpose of this study was to determine if PPARs are expressed in growth plate chondrocytes and to describe the biological effect of PPAR activation in these cells. The results demonstrate the presence of three PPAR isoforms (alpha, delta, and gamma) in growth plate cartilage. Activation of PPAR-gamma by ciglitazone in growth plate chondrocytes inhibits T(3) induced terminal differentiation and promotes apoptosis through increased levels of caspase 3/7 activity and decreased expression of the anti-apoptotic protein Bcl 2. PMID- 15878254 TI - The course of bone healing is influenced by the initial shear fixation stability. AB - Fracture healing is influenced by fixation stability and experimental evidence suggests that the initial mechanical conditions may determine the healing outcome. We hypothesised that mechanical conditions influence not only the healing outcome, but also the early phase of fracture healing. Additionally, it was hypothesised that decreased fixation stability characterised by an increased shear interfragmentary movement results in a delay in healing. Sixty-four sheep underwent a mid-shaft tibial osteotomy which was treated with either a rigid or a semi-rigid external fixator. Animals were sacrificed at 2, 3, 6 and 9 weeks postoperatively and the fracture callus was analysed using radiological, biomechanical and histological techniques. The tibia treated with semi-rigid fixation showed inferior callus stiffness and quality after 6 weeks. At 9 weeks, the calluses were no longer distinguishable in their mechanical competence. The calluses at 9 weeks produced under rigid fixation were smaller and consisted of a reduced fibrous tissue component. These results demonstrate that the callus formation over the course of healing differed both morphologically and in the rate of development. In this study, we provide evidence that the course of healing is influenced by the initial fixation stability. The semi-rigid fixator did not result in delayed healing, but a less optimal healing path was taken. An upper limit of stability required for successful healing remains unknown, however a limit by which healing is less optimal has been determined. PMID- 15878255 TI - Expression of leptin and its receptors in various tissues of ruminants. AB - The energy metabolism of domestic animals is under the control of hormonal factors, which include thyroid hormones and leptin. Leptin signals from the periphery to the centre. It is mostly produced in the white adipose tissue and informs the central nervous system (CNS) about the total fat depot of the body. Low and high levels of leptin induce anabolic and catabolic processes, respectively. Besides controlling the food uptake and energy expenditure leptin is also involved in regulation of the reproduction and the immune system. Leptin is produced in several tissues other than fat. In the present paper the leptin expression of ruminant (Egyptian water buffalo, cow, and one-humped camel) tissues are examined. The mammary gland produces leptin in each species investigated. The local hormone production contributes to milk leptin and most probably helps to maintain lactation. Considerable leptin receptor expression was observed in the milk-producing epithelial cells, which is the same cell type that produces most of the udder leptin. Based on the results tissues participating in production have an autoregulative mechanism through which tissues can be relatively independent of the plasma leptin levels in order to maintain the desired function. PMID- 15878256 TI - The effect of feeding strategy during the pre-follicular phase on subsequent follicular development in the pig. AB - In female pigs feeding level has important effects on reproductive performance. This review is focused on the follicular development after low and high feeding levels during the luteal phase in gilts. Although aspects of diet composition seem to have a role in regulating reproductive performance, the most important aspect appears to be the plane of nutrition. Similar effects are described during lactation in primiparous sows, when their metabolism is challenged to its maximum. Also in this situation feeding level clearly affects weaning to oestrus interval, follicular development, oocyte maturation, ovulation rate and subsequent embryonic survival. PMID- 15878257 TI - An experimental evaluation of ablation devices for the local treatment of the liver resection edge. AB - Surgical margin is the only technical variable that has an impact on long-term outcome after liver resection. In this study we compared radiofrequency (RFA), cryotherapy, diathermy and argon beam for the local treatment of liver resection edge. METHODS: Sixty eight ablations were produced on the surface of ex vivo sheep liver using the various modalities. Cryoablations were produced using both the Cryotech and the Erbe probes. Radiofrequency ablations were produced using a custom-made surface application probe and the RITA 1500 generator. Conmed 7500 system was used to produce diathermy and argon beam coagulation. RESULTS: Argon beam and the diathermy resulted in ablation to maximum depth of 3.5 mm with endpoint and spray modes at various power setting. RF ablation resulted in consistent ablations the diameter of which varied in a linear manner to the time of RF application and the depth of the ablations with the length of electrode deployment. Cryotherapy was as effective as RFA with both the cryotherapy systems but the Erbe trocar probe resulted in a deeper ablation, whereas the Cryotech paddle probe resulted in a larger diameter ablation. CONCLUSIONS: RF and cryotherapy are equally effective as liver edge ablation device. Diathermy and argon are considered less effective. Cryotherapy requires expensive complex equipment that at least with liquid nitrogen systems requires to be prepared for use and this may not be available if the need for edge treatment during resection was unplanned. PMID- 15878258 TI - The management of elderly patients with T1-T2 breast cancer treated with or without radiotherapy. AB - AIM: The aim of the current study is to identify a subgroup of patients with breast cancer who have a low risk of local recurrence after conservative surgery in order to avoid radiotherapy treatment. METHODS: A group of 472 patients underwent conservative surgery without radiotherapy, and it was compared to a second group of 755 patients with similar characteristics, but who had received radiotherapy treatment (RT) after conservative surgery. RESULTS: Breast relapse's univariate analysis demonstrated statistical significance for the following factors: radiotherapy treatment, clinical stage, pathological stage, positive axillary nodes and tumour grading. Different results were obtained studying breast relapse. In the no-RT group breast relapse was 10.6% while in the irradiated group it was 3.4%. The breast relapse incidence decreases as the age of the patients increases especially over 75 years of age. CONCLUSIONS: In conclusion, there is clinical evidence of avoiding adjuvant radiotherapy for patients over 75 years with T1-T2 cancer treated with quadrantectomy with a clear excision margin. PMID- 15878259 TI - Influence of hospital volume on local recurrence and survival in a population sample of rectal cancer patients. AB - AIMS: To investigate the role of hospital volume and individual hospitals on long term outcomes (local recurrence and survival) of rectal cancer patients. METHODS: One thousand thirty-eight patients with rectal cancer were diagnosed between 1996 and 1998. From these, we analysed 884 patients with a resected invasive primary rectal cancer. Median follow-up was 5.7 years. The impact of hospital volume (<10, 10-30 and >30 rectal cancer patients annually) on local recurrence and survival was examined in a Cox model. Differences between the four largest clinics in the high volume group were also investigated. RESULTS: In the multivariate model predicting survival the following variables were significant: UICC stage, grade, age, local recurrence, and (neo-) adjuvant therapy treatment. In the multivariate model predicting local recurrence UICC stage, tumour localisation, and neoadjuvant therapy treatment were significant variables. Hospital volume was not a significant factor for survival or local recurrence. Within the high volume category one hospital showed significantly worse local recurrence rates than all other hospitals, but no survival difference could be seen between the four largest hospitals of the high volume group. CONCLUSIONS: This analysis of a rectal cancer population found that hospital volume did not determine survival or local recurrence. Detailed clinical data with long term follow-up from cancer registries are vital to demonstrate the quality of routine care. PMID- 15878260 TI - Surgical quality assurance in rectal cancer treatment: the key to improved outcome. AB - Surgery is the key to cure for patients with rectal cancer. Surgical techniques are evolving with conventional blunt dissection being increasingly abandoned for the sake of total mesorectal excision (TME), leading to favourable clinical and functional outcome. Surgical quality assurance programmes, involving training of general surgeons to adopt the TME technique, have proven to be feasible and result in improved outcome compared to historical controls. In this overview, developments in rectal cancer treatment are highlighted, the relation of surgeon and/or hospital volume with treatment outcome is discussed, and future directions in optimising rectal cancer treatment are considered. PMID- 15878261 TI - Tofupill lacks peripheral estrogen-like actions in the rat reproductive tract. AB - OBJECTIVE: The objective of this study was to evaluate the estrogenic effect of phytoestrogens contained in a commercial food supplement (Tofupill) on the reproductive tract of ovariectomized rats. METHODS: Food supplement (3.4 or 10.2 mg/kg) and conjugated equine estrogens (CEE, 31 or 100 microg/kg) were orally administered, daily during 14 days to ovariectomized rats. At the end of treatment, the following determinations were done: dry and wet uterine weight, vaginal epithelium condition, and uterine serotonin-induced contractile response. A group treated with 17beta-estradiol was included as control for serotonin induced contractile response. RESULTS: Food supplement did not display clear estrogenic effects on vaginal epithelium, uterine weight or myometrial sensitivity to serotonin, whereas high doses of conjugated equine estrogens showed estrogenic action. CONCLUSIONS: The present data showed that Tofupill displayed a lower estrogenic effect than conjugated equine estrogens, which are one of the most commonly used hormone replacement therapy for postmenopausal women. However, further studies are needed to evaluate the risk associated to the use of Tofupill as an alternative to hormone replacement therapy for postmenopausal women. PMID- 15878262 TI - Predictive factors of lopinavir/ritonavir discontinuation for drug-related toxicity: results from a cohort of 416 multi-experienced HIV-infected individuals. AB - The objective of this study was to find predictive factors of lopinavir/ritonavir (LPV/r) discontinuation for drug-related toxicities in highly pre-treated human immunodeficiency virus (HIV)-infected subjects. The study was an observational study of HIV patients starting LPV/r with HIV RNA > 3log10 copies/mL and a follow up > or = 6 months. Parameters studied were HIV RNA, CD4+ cell counts, metabolic parameters and drug-related adverse events. Acquired immune deficiency syndrome (AIDS) events and deaths were recorded. The Kaplan-Meier (KM) model was used to estimate time-dependent probability, and the multivariable Cox model to identify predictors of LPV/r discontinuation for adverse events. The study evaluated 416 HIV-infected patients. Seventy-seven patients (18.5%) discontinued LPV/r for toxicities. Adverse events leading to LPV/r discontinuation were gastrointestinal symptoms in 40 cases, hyperlipidaemia in 27 and increase of aspartate aminotransferase (AST)/alanine aminotransferase (ALT) in 10 patients. Nineteen patients (4.6%) developed an AIDS event during observation and 15 (3.6%) died. The KM probability of LPV/r discontinuation for toxicities was 5.3% (range 3.1 7.5%) at month 12 and 15.7% (range 12.1-19.3%) at month 24. Subjects with hepatitis C virus (HCV)-HIV co-infection (odds ratio (OR) 7.40; 95% confidence interval (CI) 3.73-14.66 versus HCV-negative; P = 0.001) and receiving LPV/r plus nucleoside reverse transcriptase inhibitors (NRTIs) and protease inhibitor (PI)/non-nucleoside reverse transcriptase inhibitor (NNRTI) (OR 1.74; 95% CI 1.04 2.91 versus LPV/r plus only NRTIs; P = 0.04) showed a higher risk of LPV/r discontinuation by a Cox analysis, whereas non-intravenous drug abusers (IVDUs) (OR 0.40; 95% CI 0.24-0.67 versus IVDUs; P = 0.001) had a lower risk. The rate of discontinuation for toxicity decreased by 17% for each additional month of LPV/r exposure (OR 0.83; 95% CI 0.80-0.86 for each additional month; P < 0.001). LPV/r was substantially well tolerated. Diarrhoea was the most frequent adverse event leading to discontinuation. HCV-HIV co-infected patients and patients with a short exposure to LPV/r have a higher risk of discontinuing LPV/r and should be strictly monitored. PMID- 15878263 TI - Molecular characterisation of Hungarian macrolide-resistant Streptococcus pneumoniae isolates, including three highly resistant strains with the mef gene. AB - The macrolide resistance of 304 Hungarian Streptococcus pneumoniae isolates was investigated. Antibiotic sensitivity testing was performed in air and in 5% CO(2). More erythromycin resistance was noted when growth was in CO(2). A resistance determinant was found in almost all isolates: erm(B) gene (87.4%), mef genes (9.2%) and one strain with the erm(TR) gene. This indicates that screening for carriage of resistance determinants should always be done in the presence of 5% CO(2). We found three isolates with mef(E), which were highly resistant to erythromycin. These contained multiple and some novel, ribosomal mutations. The most prevalent serogroups were 6, 19 and 14. Based on the PFGE pattern, we found identity between the Hungarian isolates and two PMEN clones. PMID- 15878264 TI - Continuous infusion of ticarcillin-clavulanate for home treatment of serious infections: clinical efficacy, safety, pharmacokinetics and pharmacodynamics. AB - Continuous infusion (CI) ticarcillin-clavulanate is a potential therapeutic improvement over conventional intermittent dosing because the major pharmacodynamic (PD) predictor of efficacy of beta-lactams is the time that free drug levels exceed the MIC. This study incorporated a 6-year retrospective arm evaluating efficacy and safety of CI ticarcillin-clavulanate in the home treatment of serious infections and a prospective arm additionally evaluating pharmacokinetics (PK) and PD. In the prospective arm, steady-state serum ticarcillin and clavulanate levels and MIC testing of significant pathogens were performed. One hundred and twelve patients (median age, 56 years) were treated with a CI dose of 9.3-12.4g/day and mean CI duration of 18.0 days. Infections treated included osteomyelitis (50 patients), septic arthritis (6), cellulitis (17), pulmonary infections (12), febrile neutropenia (7), vascular infections (7), intra-abdominal infections (2), and Gram-negative endocarditis (2); 91/112 (81%) of patients were cured, 14 (13%) had partial response and 7 (6%) failed therapy. Nine patients had PICC line complications and five patients had drug adverse events. Eighteen patients had prospective PK/PD assessment although only four patients had sufficient data for a full PK/PD evaluation (both serum steady state drug levels and ticarcillin and clavulanate MICs from a bacteriological isolate), as this was difficult to obtain in home-based patients, particularly as serum clavulanate levels were found to deteriorate rapidly on storage. Three of four patients with matched PK/PD assessment had free drug levels exceeding the MIC of the pathogen. Home CI of ticarcillin-clavulanate is a safe, effective, convenient and practical therapy and is a therapeutic advance over traditional intermittent dosing when used in the home setting. PMID- 15878265 TI - Electrophysiological ratio markers for the balance between reward and punishment. AB - It has been argued that prototypical forms of psychopathology result from an imbalance in reward and punishment systems. Recent studies suggest that the ratios between slower and faster waves of the electroencephalogram (EEG) index this motivational balance and might therefore have diagnostic value for psychopathology. To scrutinize this notion, the present study investigated whether resting state EEG ratios would predict decision making on the Iowa gambling task (Iowa-GT), a well-known marker for motivational imbalance. A resting state EEG recording was acquired followed by the Iowa-GT in twenty-eight healthy right-handed volunteers. Results showed that higher versus lower EEG ratios were associated with disadvantageous versus advantageous decision making strategies indicating motivational imbalances in reward- and punishment-driven behavior, respectively. This finding provides the first direct evidence that the electrophysiologically derived EEG ratios can serve as biological markers for balance and imbalance in motivation. PMID- 15878266 TI - Nuclear type II [3H]estradiol binding sites: a histone H3-H4 complex. AB - [(3)H]luteolin covalently labels two forms (11kDa and 35kDa proteins) of type II binding sites in rat uterine nuclear extracts [K. Shoulars, T. Brown, M. Alejandro, J. Crowley, B. Markaverich, Identification of rat uterine nuclear type II [(3)H]estradiol binding sites as histone H4, Biochem. Biophys. Res. Commun. 296 (2002) 1083-1090]. The 11kDa protein was identified as histone H4. Levels of the 35kDa protein were insufficient for sequencing; however, this protein was recognized by anti-histone H4 antibodies. Histones H3 and H4 exist as dimers in vivo (mw>>35kDa) and we suspected the 35kDa [(3)H]luteolin-labeled protein in uterine nuclear extracts might be a complex of histones H3 and H4. This manuscript describes methods for the purification of commercially available calf thymus core histones that retain [(3)H]luteolin binding activity and are of sufficient purity for recombination studies. Mixing experiments with pure H3 and H4 from calf thymus demonstrate that a 35kDa H3-H4 dimer capable of binding [(3)H]luteolin is generated and this protein appears equivalent to the 35kDa [(3)H]luteolin binding protein in rat uterine nuclear extracts. If this is the case, type II site ligands including MeHPLA, luteolin, and other bioflavonoids and phytoestrogens may control histone-dependent gene transcription and cellular proliferation via binding to and modulating core histone/nucleosome function. PMID- 15878267 TI - A strategy to profile prime and non-prime proteolytic substrate specificity. AB - A strategy was developed to determine the prime and non-prime substrate specificity of serine, threonine and cysteine proteases. ACC positional scanning technology was employed to determine the P4-P1 non-prime site substrate specificity. The data was used to synthesize biased donor-quencher positional scanning libraries to profile the P1'-P4' prime site substrate specificity. Directed sorting using the Irori Nanokan system allowed for the archiving of multiple P1'-P4' positional scanning libraries. From these libraries focused donor-quencher libraries incorporating P4-P1 data for each protease under study could be rapidly prepared. The profiling of thrombin and caspase-3 P4-P4' substrate specificity, comparison of the library specificity data to single substrates, and the analysis of physiological cleavage sites are described. PMID- 15878268 TI - Antioxidant and antiproliferative activity of curcumin semicarbazone. AB - A new semicarbazone derivative of curcumin (CRSC) was synthesized and examined for its antioxidant, antiproliferative, and antiradical activity and compared with those of curcumin (CR). The antioxidant activity was tested by their ability to inhibit radiation induced lipid peroxidation in rat liver microsomes. The antiproliferative activity was tested by studying the in vitro activity of CRSC against estrogen dependant breast cancer cell line MCF-7. Kinetics of reaction of (2,2'-diphenyl-1-picrylhydrazide) DPPH, a stable hydrogen abstracting free radical was studied to measure the antiradical activity using stopped-flow spectrophotometer. Finally one-electron oxidized radicals of CRSC were generated and characterized by pulse radiolysis. The results suggest that the probable site of attack for CRSC is both the phenolic OH and the imine carbonyl position. CRSC shows efficient antioxidant and antiproliferative activity although its antiradical activity is less than that of CR. PMID- 15878269 TI - A switch in enantiomer preference between mitochondrial F1F0-ATPase chemotypes. AB - The preferred absolute configuration of two series of F(1)F(0)-ATP synthase inhibitors was determined. Although the configuration of the active enantiomer in each series is different, each series presents the same 'triaryl' pharmacophore to the enzyme binding site. PMID- 15878270 TI - Chiral discrimination in binding of enantiomers of 2-(aminoalkoxy)-substituted 4 (2-thienyl)pyrimidines and 4,6-bis(2-thienyl)pyrimidines with duplex DNA. AB - Thienylpyrimidines substituted at position 2 of the pyrimidine with a chiral aminoalkoxy group were synthesized. Upon interaction with duplex DNA, the unfused heteroaromatic system of these compounds intercalates with DNA base pairs and the protonated side chain is located in the major groove. The S-enantiomers bind more strongly than their R-counterparts with enantiomeric discrimination, as measured by a ratio of binding constants K(S)/K(R), ranging from 1.2 to 2.4. PMID- 15878271 TI - Tethered phytic acid as a probe for measuring phytase activity. AB - A novel approach for measuring phytase activity is presented. We have developed a new chromophoric substrate analog of phytic acid, 5-O-[6-(benzoylamino)hexyl]-d myo-inositol-1,2,3,4,6-pentakisphosphate that permits direct measurement of the phosphate ester bond-cleavage reaction using HPLC. This compound, along with its dephosphorylated T-phosphatidylinositol intermediates, are quantified using reversed phase chromatography with UV detection. PMID- 15878272 TI - Antimalarial activity and synthesis of new trisubstituted pyrimidines. AB - A series of 2,4,6-trisubstituted-pyrimidines was synthesized and evaluated for their in vitro antimalarial activity against Plasmodium falciparum. Out of the 30 compounds synthesized 21 compounds showed MIC in the range of 0.5-2 microg/mL. These compounds are in vitro several folds more active than pyrimethamine. PMID- 15878273 TI - Total asymmetric synthesis of (-)-conduramine B-1 and of its enantiomer. N-Benzyl derivatives of conduramine B-1 are beta-glucosidase inhibitors. AB - The 'naked sugars' (+)- and (-)-7-oxabicyclo[2.2.1]hept-5-en-2-one have been converted into (-)-conduramine B-1 ((-)-3) and its enantiomer (+)-3, respectively. They have been condensed with a variety of aldehydes in the presence of NaBH(OAc)(3). The N-substituted derivatives 4 and ent-4 so-obtained have been tested against two alpha-glucosidases, two amyloglucosidases, two beta glucosidases and one beta-xylosidase for their inhibitory activities. Although ( )-3 and (+)-3 do not inhibit any of these enzymes at 1mM concentration, N benzylated derivatives of (-)-conduramine B-1 are selective and competitive inhibitors of beta-glucosidases with K(i) in low micromolecular range. PMID- 15878274 TI - Discovery of potent and use-dependent sodium channel blockers for treatment of chronic pain. AB - A new series of voltage-gated sodium channel blockers with potential for treatment of chronic pain is reported. Systematic structure-activity relationship studies, starting with compound 1, led to identification of potent analogs that displayed use-dependent block of sodium channels, were efficacious in pain models in vivo, and most importantly, were devoid of activity against the cardiac potassium channel hERG. PMID- 15878275 TI - Synthesis of tricyclic analogs of stephaoxocanidine and their evaluation as acetylcholinesterase inhibitors. AB - The synthesis of simplified analogs of the novel isoquinoline alkaloid stephaoxocanidine, carrying the oxazaphenalene ABC-ring system of the natural product, and their activity as inhibitors of the enzyme acetylcholinesterase, are reported. 5,6-Dimethoxy-7H -8-oxa-1-aza-phenalen-9-one (5) was as active as a Narcissus extract enriched in galantamine. PMID- 15878276 TI - Inhibition of hydrogen peroxide-induced necrotic cell death with 3-amino-2 indolylmaleimide derivatives. AB - Novel analogs of indolylmaleimide derivatives (IM derivatives) were synthesized and tested for cell death-inhibitory activity. 2-(1H-Indol-3-yl)-3-pentylamino maleimide IM-54 was the most effective cell death inhibitor among the compounds tested. IM-54 inhibited necrotic cell death induced by H2O2, but not apoptotic cell death induced by etoposide. These results indicated that this novel cell death inhibitor is distinct from the well-known caspase inhibitor, Z-VAD, which can block apoptotic cell death, but not necrotic cell death. IM-54 is expected to be a powerful bioprobe for clarifying the unique signaling pathway of necrotic cell death. PMID- 15878277 TI - Synthesis and phosphodiesterase 5 inhibitory activity of novel pyrido[1,2-e]purin 4(3H)-one derivatives. AB - Synthesis and primary SAR of a novel series of 2-phenylpyrido[1,2-e]purin-4(3H) one derivatives with piperazinyl sulfonamide substituents were described herein. As potential PDE5 inhibitors for erectile dysfunction (ED) treatment, representative compounds exhibit improved selectivity versus PDE1 and PDE6. Meanwhile, compound 3e demonstrated functional efficacy on rabbit corpus cavernosum strip in vitro. PMID- 15878278 TI - The evaluation of 2,8-disubstituted benzoxazinone derivatives as anti inflammatory and anti-platelet aggregation agents. AB - A series of 2,8-disubstituted benzoxazinones were synthesized and subjected to anti-platelet aggregation, inhibition of superoxide anion generation, and inhibition of neutrophil elastase release assays. Among them, 2-(2'-substituted phenyl)-benzoxazinones exhibited significant inhibitory effect to target assays. Additionally, all of them were more potent than aspirin on AA-induced platelet aggregation, and these suggested that 2-(2'-substituted-phenyl)-benzoxazinones also possess aspirin-like activity. On the other hand, the compounds 6 and 16 showed inhibitory effects on neutrophil elastase release and superoxide generation. PMID- 15878279 TI - A-ring hydroxymethyl 19-nor analogs of the natural hormone 1alpha,25 dihydroxyvitamin D(3): synthesis and preliminary biological evaluation. AB - A series 5-8 of 1- and 3-CH(2)OH 19-nor analogs of the hormone calcitriol (1) has been prepared. Surprisingly, 19-nor 1alpha-CH(2)OH analog 5a is more antiproliferative at 100 nM concentration than the corresponding regioisomeric analog 6a with the natural 1alpha-OH group, and 1alpha-CH(2)OH hybrid analog 7a is similar in antiproliferative potency to calcitriol (1) even at low nanomolar concentrations. PMID- 15878280 TI - A prenylbisabolane with NF-kappaB inhibiting properties from Cascarilla (Croton eluteria). AB - Investigation of the bark of Croton eluteria Bennett for biologically active compounds has led to the isolation of the new prenylbisabolane 3, whose structure was assessed by spectroscopic methods. The corresponding known enone 4 and the eudesmane sesquiterpene 2 were also obtained. Compound 3 proved active in selectively inhibiting the induction of NF-kappaB by tumor necrosis factor-alpha in T cells. PMID- 15878281 TI - Biomimetic synthesis of acid-sensitive (-)- and (+)-caparrapi oxides, (-)- and (+)-8-epicaparrapi oxides, and (+)-dysifragin induced by artificial cyclases. AB - Asymmetric total syntheses of acid-sensitive (-)- and (+)-caparrapi oxides (1) and (+)-8-epicaparrapi oxide (2) from farnesol (10) are achieved using Sharpless Katsuki epoxidation and Lewis acid-assisted chiral Bronsted acid (chiral LBA) induced polyene cyclization as key steps. The relative configuration of (+) dysifragin (4) is determined by a single-crystal X-ray diffraction and its total synthesis is accomplished by the diastereoselective epoxidation of (+)-1. Furthermore, (-)-1 can be directly synthesized from (S)-nerolidol (3) and (R)-LBA with 88% ds by reagent control, which overcame substrate control, while (-)-2 is obtained from (R)-3 and (R)-LBA with >99% ds by the double asymmetric induction. PMID- 15878282 TI - 3D-QSAR study of bis-azaaromatic quaternary ammonium analogs at the blood-brain barrier choline transporter. AB - Previously, we have developed 3D-QSAR models of the blood-brain barrier (BBB) choline transporter, a transport system that may have utility as a vector for central nervous system drug delivery. In this study, we extended the model by evaluating five bis-azaaromatic quaternary ammonium compounds for their affinity for the choline binding site on the BBB-choline transporter. The compounds, and their affinities for the transporter, were then incorporated into our existing molecular model, in order to update our knowledge on the molecular recognition factors associated with interaction of ligands at the choline binding site. The current compounds are structurally related to previous substrates that we have evaluated, but offer additional three dimensional aspects compared to the series of compounds previously utilized to define the original models. The compounds showed good affinity for the BBB-choline transporter, exhibiting inhibition constants ranging from 10 to 68 microM, as determined by the in situ rat brain perfusion method. Comparative molecular field analysis (CoMFA) and comparative molecular similarity index analysis (CoMSIA) methods were used to build the new 3D QSAR models. When the new bis-azaaromatic quaternary ammonium compounds were included in the model, the best cross-validated CoMFA q2 was found to be 0.536 and the non-cross-validated r2 was 0.818. CoMSIA hydrophobic cross-validated q2 was 0.506 and the non-cross-validated r2 was 0.804. This new model was able to better predict BBB-choline transporter affinity of hemicholinium-3 (predicted 65 microM, actual 54 microM), when compared to an earlier model (predicted 316 microM). PMID- 15878283 TI - Synthesis of gossypol atropisomers and derivatives and evaluation of their anti proliferative and anti-oxidant activity. AB - Gossypol 1, gossypolone 2, and a series of bis 3 and half Schiff's bases 4 of gossypol were synthesised and tested for anti-proliferative and anti-oxidant activity. (-)-Gossypol (-)-1 was the most potent inhibitor of the proliferation of the HPV-16 keratinocyte cell line (using an MTT viability assay) with a GI50 of 4.8 microM. The bis Schiff's base of (-)-gossypol with L-tyrosine ethyl ester (-)-3b was the most potent inhibitor of iron/ascorbate dependent lipid peroxidation (using the thiobarbituric acid test), with an IC50 of 11.7 microM, with (-)-gossypol being the next most potent of the series, with an IC50 of 13.1 microM. The results from these initial assays suggest that gossypol, as either a racemic mixture rac-1, or the individual atropisomers (-)-1 or (+)-1, has potential for the treatment of psoriasis. PMID- 15878284 TI - CO(2)-induced total phenolics in suspension cultures of Panax ginseng C. A. Mayer roots: role of antioxidants and enzymes. AB - The effects of different concentrations of CO(2) (1%, 2.5% and 5%) on the antioxidant capacity, total phenols, flavonoids, protein content and phenol biosynthetic enzymes in roots of Panax ginseng were studied in bioreactor (working volume 4 l) after 15, 30 and 45 days. CO(2) induced accumulation of total phenolics in a concentration and duration dependent manner. Total phenols, flavonoids and 1,1-diphenyl-2-picrylhydrazyl (DPPH) activity increased 60%, 30% and 20% at 2.5% CO(2) after 45 days compared to control in P. ginseng roots which indicated that phenolics compounds played an important role in protecting the plants from CO(2). Hypothesizing that increasing the phenolic compounds in roots of P. ginseng may increase its nutritional functionality; we investigated whether pentose phosphate pathway (PPP), shikimate/phenylpropanoid pathway enzymes have a role in phenolics mobilization in P. ginseng roots. Fresh weight (FW), dry weight (DW) and growth ratio was increased at 1% and 2.5% CO(2) only after 45 days, however, unaffected after 15 and 30 days. Results also indicated that high CO(2) progressively stimulated the activities of glucose 6 phosphate dehydrogenase (G6PDH, E.C. 1.1.1.49), shikimate dehydrogenase (SKDH, E.C. 1.1.1.25), phenylalanine ammonia lyase (PAL, E.C. 4.3.1.5), cinnamyl alcohol dehydrogenase (CAD, E.C. 1.1.1.195), caffeic acid (CA) peroxidase and chlorogenic acid (CGA) peroxidase after 15, 30 and 45 days. Increased CO(2) levels resulted in increases in accumulation of total protein (45%), non-protein thiol (NP-SH) (30%) and cysteine contents (52%) after 45 days compared to control and increased activities of beta-glucosidase (GS, E.C. 3.2.1.21) and polyphenol oxidase (PPO, E.C. 1.10.3.2) in P. ginseng roots indicated that they played an important role in protecting the plants from CO(2). These results strongly suggest that high concentration of CO(2) delivered to ginseng root suspension cultures induced the accumulation of total phenolics possessing high antioxidant properties probably useful for human health. Therefore, roots of P. ginseng are considered as a good source of phenolics compounds with high antioxidants capacity and can be produced on a large scale. PMID- 15878285 TI - Association of interleukin-1B (IL-1B) gene polymorphisms with risk of gastric cancer in Chinese population. AB - The incidence of gastric cancer (GC) in China is among the highest in the world. In present work, 154 patients with GC and 166 healthy controls in population of north-western China were investigated to evaluate the genetic associations of IL 1B gene single nucleotide polymorphisms (SNP) and variable number tandem repeat (VNTR) polymorphisms of IL-1RN gene with increased risk of GC. The frequency of IL-1B+3954C/T was significantly higher in GC cases group (25.97%) than that in controls (4.82%) with odds ratio (OR)=6.93 (95% confidence interval [CI] 3.13 15.36); the frequencies of IL-1B-31C/T, IL-1B-31C/C and IL-1B-511C/T genotypes were also higher in GC cases group (51.95%, 23.38% and 50.65%) than those in controls (46.99%, 19.88% and 42.77%) with OR=1.48 (95% CI 0.88-2.49), OR=1.58 (95% CI 0.84-2.95) and OR=1.39 (95% CI 0.80-2.41), respectively. The results show that these SNPs of IL-1B gene are associated with significantly increased risk of GC. This is the first report that IL-1B+3954C/T heterozygote is associated with greatly increased risk of GC. The results of this study did not support the report that IL-1RN*2+ genotypes were associated with increased risk of GC in Chinese population. PMID- 15878286 TI - The structure of gas-phase bradykinin fragment 1-5 (RPPGF) ions: an ion mobility spectrometry and H/D exchange ion-molecule reaction chemistry study. AB - Ion mobility-mass spectrometry (IM-MS) data is interpreted as evidence that gas phase bradykinin fragment 1-5 (BK1-5, RPPGF) [M + H](+) ions exist as three distinct structural forms, and the relative abundances of the structural forms depend on the solvent used to prepare the matrix-assisted laser desorption ionization (MALDI) samples. Samples prepared from organic rich solvents (90% methanol/10% water) yield ions having an ion mobility arrival-time distribution (ATD) that is dominated by a single peak; conversely, samples prepared using mostly aqueous solvents (10% methanol/90% water) yield an ATD composed of three distinct peaks. The BK1-5 [M + H](+) ions were also studied by gas-phase hydrogen/deuterium (H/D) exchange ion-molecule reactions and this data supports our interpretation of the IM-MS data. Plausible structures for BK1-5 ions were generated by molecular dynamics (MD). Candidate MD-generated structures correlated to measured cross-sections suggest a compact conformer containing a beta-turn whereas a more extended, open form does not contain such an interaction. This study illustrates the importance of intra-molecular interactions in the stabilization of the gas-phase ions, and these results clearly illustrate that solution-phase parameters (i.e., MALDI sample preparation) greatly influence the structures of gas-phase ions. PMID- 15878287 TI - The use of correlation integrals in the study of localized muscle fatigue of elbow flexors during maximal efforts. AB - Innovative applications of non-linear time series analysis have recently been used to investigate physiological phenomena. In this study, we investigated the feasibility of using the correlation integral to monitor the localized muscle fatigue process in the biceps brachii during sustained maximal efforts. The subjects performed isometric maximum contractions until failure in elbow flexion (90 degrees from neutral). The median and the 70th percentile frequency of the Surface electromyography (SEMG) power spectrum, the integrated SEMG, and the Correlation Integral (CI) were evaluated during the trials. The linear correlation between these variables and the elbow torque production was used to quantify the ability of a parameter to follow the fatiguing process. The CI had the highest linear correlation with torque (0.77 (0.12SD)), while the spectral indices correlations with torque were much lower. The decreasing trend of the torque production was followed by the spectral indices for only the beginning part of the contraction, while the CI increased sharply after the torque production fell to about 0.60 of the MVC. This suggests that the CI is sensitive to different changes of the SEMG signal during fatigue than the spectral variables. PMID- 15878288 TI - Statistical modelling of fatigue-related electromyographic median frequency characteristics of back and hip muscles during a standardized isometric back extension test. AB - The purpose of the present study was to evaluate which statistical model - linear, logarithmic, quadratic or exponential - best described the fatigue related electromyographic (EMG) changes of back and hip muscles. Twenty healthy volunteers performed a modified Biering-Sorensen test. The EMG activity of the latissimus dorsi (LD), longissimus thoracis pars thoracis (LTT) and lumborum (LTL), iliocostalis lumborum pars thoracis (ILT) and lumborum (ILL), multifidus (MF), gluteus maximus (GM) and biceps femoris (BF) was measured bilaterally using surface electrodes. Higher R(2) values were found for the quadratic models (p<0.05 for all muscles), and lower R(2) values for the logarithmic models (p<0.05 for LTT, LTL, ILL, MF and GM). The exponential models generated higher R(2) values compared to the linear ones for the LTT, LTL and MF (all p<0.05). Further analyses revealed, however, that these models did not add useful additional information, and therefore would only increase the complexity. The findings of the current study validate the use of simple linear regression techniques when studying fatigue-related EMG median frequency characteristics of back and hip muscles during isometric contractions. PMID- 15878289 TI - Analysis on the activation of the VMO and VLL muscles during semisquat exercises with and without hip adduction in individuals with patellofemoral pain syndrome. AB - OBJECTIVE: The purpose of this study was to investigate the effect of hip adduction on the activity of the Vastus Medialis Obliquus (VMO) and Vastus Lateralis Longus (VLL) muscles during semisquat exercises. METHODS: Twenty female subjects, divided into two groups comprising healthy and patellofemoral pain syndrome (PFPS) subjects (ten volunteers for each group), performed three double leg semisquat exercise trials with maximum hip adduction isometric contraction (DLSS-HA) and three double-leg semisquat exercise trials without hip adduction (DLSS). The normalized electromyographic muscle data were analysed using Repeated Measure ANOVA (p or =10), allocated using EEG linked video-recording of concurrent behaviour, and sensitivity and specificity values had to be stated or be calculable. RESULTS: Thirty-three papers were identified, of which 13 satisfied criteria. Excluded studies are briefly described. Those retained comprised a range of procedures [seizure induction, MMPI assessment, physiological assessment (prolactin, SPECT), pre-ictal pseudosleep, and ictal/post-ictal characteristics]. No procedure emerged with both high sensitivity and specificity and adequately replicated findings, although high levels of specificity were more commonly reported than high levels of sensitivity. This suggests that procedures were generally better at excluding a possible diagnosis. CONCLUSIONS: No procedure attains reliability equivalent to EEG video-telemetry. Further rigorous evaluation, using standardised and replicable methodologies, is required. The range of symptoms presented in NES suggests that a multi-method approach may be required. This too would require evaluation. PMID- 15878292 TI - Did Fyodor Mikhailovich Dostoevsky suffer from mesial temporal lobe epilepsy? AB - Many scientific authors--among them famous names such as Henri Gastaut or Sigmund Freud--dealt with the question from what kind of epilepsy Fyodor Mikhailovitch Dostoevsky (1821-1881) might had suffered. Because of the tight interplay between Dostoevsky's literary work and his own disease we throw light on the author's epilepsy against the background of his epileptic fictional characters. Moreover, we attempt to classify Dostoevsky's epilepsy on the basis of his bibliography, language, and literary work. PMID- 15878293 TI - Expression of fibroblast growth factor receptor 3 by fibroblast growth factor 2 in cultured chick embryo chondrocytes. AB - Although fibroblast growth factor 2 (FGF2) and fibroblast growth factor receptor 3 (FGFR3) both inhibit longitudinal bone growth, little is known about the relationship between FGF2 and FGFR3. Accordingly, the current study examined the expression of FGFR3 mRNA after the administration of FGF2 using cultured chondrocytes from day 17 chick embryos to evaluate the relationship between FGF2 and FGFR3. The chondrocytes were isolated from the caudal one-third portion (LS) of sterna, peripheral regions (USP) and central core regions (USC) of the cephalic portion of the sterna, and lower portion of the proximal tibial growth plate (Ti) of day 17 chick embryo. The expression of FGFR1, FGFR3, and type II and X collagen mRNA in the chondrocytes from the LS, USP, USC, and Ti was determined. FGFR1 was not expressed in the LS and USP chondrocytes, yet strongly expressed in the USC and Ti chondrocytes. With a treatment of FGF2, the expression of FGFR1 slightly increased in the USC chondrocytes and was not related with the concentration of FGF2 in the Ti chondrocytes. FGFR3 was expressed in all the chondrocyte types, yet strongly increased in the LS, USC, USP, and Ti in that order according to the concentration of FGF2. For the LS and USP chondrocytes, the expression of FGFR3 with FGF2 increased in a 4-day culture, yet decreased in a 6-day culture, whereas for the USC chondrocytes, the expression of FGFR3 mRNA with FGF2 increased in a 2-day culture, yet decreased in a 4-day culture, suggesting that the hypertrophic chondrocytes were more numerous and sensitive compared to the proliferative chondrocytes. For all the chondrocyte types, FGF2 appeared to be up-regulated to FGFR3, as the expression of FGFR3 mRNA increased with a higher concentration of FGF2 until a peak level. In conclusion, FGF2 was found to up-regulate to FGFR3 until the peak level of FGFR3 mRNA expression, while in hypertrophic chondrocytes, FGFR3 appeared to cause the differentiaton of chondrocytes, resulting in the inhibition of longitudinal bone growth after the peak level of FGFR3 mRNA expression. PMID- 15878294 TI - Human neuroblastoma GOTO cells express CD44 and localize it into lipid rafts upon differentiation into Schwannian cells. AB - Neuroblastoma, which is a malignant tumor consisting of dedifferentiated neuroectodermal cells, is known to show spontaneous maturation or regression in its growth. Cultured human neuroblastoma GOTO cells could be induced to differentiate into Schwannian cells and neuronal cells by incubation in the presence of 5-bromo-2'-deoxyuridine (BrdU) and by serum depletion, respectively. Here we report that in association with these differentiations, cells differentiated into Schwannian cells specifically expressed a cell adhesion molecule CD44, of which expression is usually suppressed in GOTO cells. In contrast, it remained suppressed in cells differentiated into neuronal cells. Polymerase-chain reaction revealed that the CD44 species expressed was the hemopoietic form (CD44H) with long cytoplasmic tail. Furthermore, the newly expressed CD44 in the cells was found exclusively in membrane microdomains, called lipid rafts. These data suggest that CD44 might play an important role in GOTO cells differentiated into Schwannian cells. PMID- 15878295 TI - Ejection mechanics and trajectory of the ascospores of Gibberella zeae (anamorph Fuarium graminearum). AB - Since wind speed drops to zero at a surface, forced ejection should facilitate spore dispersal. But for tiny spores, with low mass relative to surface area, high ejection speed yields only a short range trajectory, so pernicious is their drag. Thus, achieving high speeds requires prodigious accelerations. In the ascomycete Gibberella zeae, we determined the launch speed and kinetic energy of ascospores shot from perithecia, and the source and magnitude of the pressure driving the launch. We asked whether the pressure inside the ascus suffices to account for launch speed and energy. Launch speed was 34.5 ms-1, requiring a pressure of 1.54 MPa and an acceleration of 870,000 g--the highest acceleration reported in a biological system. This analysis allows us to discount the major sugar component of the epiplasmic fluid, mannitol, as having a key role in driving discharge, and supports the role of potassium ion flux in the mechanism. PMID- 15878296 TI - Fear-avoidance beliefs and pain coping strategies in relation to lower back problems in a South African steel industry. AB - The objective was to determine the association between the prevalence of lower back problems (LBP), fear-avoidance beliefs and pain coping strategies using an analytical cross-sectional epidemiological study among a group of 366 workers in a South African stainless steel industry. Outcome (LBP) was defined using a questionnaire and a functional rating index. Exposure to psychosocial risk was determined using the Fear-Avoidance Beliefs (FABQ) and Coping Strategies (CSQ) questionnaires. Multivariate logistic regression analyses for LBP indicated the following significant risk factors: work-related fear-avoidance beliefs (OR 3.40; 95% CI 2.20-5.25), catastrophizing (1.31; 1.01-1.7) and pain coping self statements (1.47; 1.16-1.87). Significant protective associations were found for increased activity levels (OR 0.57; 95% CI 0.42-0.78). These findings have utility in preventative screening procedures to identify workers with such beliefs and coping strategies who are at risk for prolonged work restrictions. PMID- 15878297 TI - A comparison of the effects of preferred music, arithmetic and humour on cold pressor pain. AB - Research studies of 'audioanalgesia', the ability of music to affect pain perception, have significantly increased in number during the past two decades. Listening to preferred music in particular may provide an emotionally engaging distraction capable of reducing both the sensation of pain itself and the accompanying negative affective experience. The current study uses experimentally induced cold pressor pain to compare the effects of preferred music to two types of distracting stimuli found effective within the previous studies; mental arithmetic, a cognitive distraction, and humour, which may emotionally engage us in a similar manner to music. Forty-four participants (24 females, 20 males) underwent three cold pressor trials in counterbalanced order. The Paced Auditory Serial Addition Task provided the cognitive distraction and a choice was given from three types of audiotaped stand-up comedy. Participants provided their own preferred music. A circulating and cooling water bath administered cold pressor stimulation. Tolerance time, pain intensity on visual analogue scale and the pain rating index and perceived control were measured. Preferred music listening was found to significantly increase tolerance in comparison to the cognitive task, and significantly increase perceived control in comparison to humour. Ratings of pain intensity did not significantly differ. The results suggest preferred music listening to offer effective distraction and enhancement of control as a pain intervention under controlled laboratory conditions. PMID- 15878298 TI - Influence of the anisotropic hyperfine interaction on the 14N ENDOR and the ESEEM orientation-disordered spectra. AB - The influence of the anisotropic hyperfine interaction on the 14N electron nuclear double resonance/electron spin echo envelope modulation spectra is studied by approximate analytical and graphical methods for the case of the isotropic g-factor. The suggested determination of the modified characteristic directions of the magnetic field due to anisotropy enhances the insight in the structural details of the system and analytical solutions of the secular equation for these conditions are derived. The graphical method, previously used for the analysis of the orientation dependence of the 14N nuclear-transition frequencies in orientation-disordered samples for isotropic hyperfine interaction is extended to the case of arbitrary anisotropic hyperfine tensor. The above analytical and graphical methods are illustrated and tested against exact simulations in two practically important cases: (i) isotropic hyperfine interaction (hfi) exceeding other nuclear interactions in nuclear spin Hamiltonian. (ii) Cancellation of the isotropic part of the hfi. PMID- 15878299 TI - Three forms of cyclin B transcripts in the ovary of the kuruma prawn Marsupenaeus japonicus: their molecular characterizations and expression profiles during oogenesis. AB - Cyclin B is a well known regulatory factor that plays a crucial role in mitosis and meiosis. Although the existence of cyclin B has been reported to be universal in a wide variety of eukaryotic organisms, no molecular data are available on crustacean species. In this study, three forms of cyclin B transcripts were first identified and characterized in the ovary of the commercially important kuruma prawn Marsupenaeus japonicus. The three transcripts (2.4, 1.9 and 1.7 kb) shared the identical sequence, with variations only in the length of 3' untranslated regions (UTRs), and coexisted in the ovary as demonstrated by Northern blot analysis. The sequences of 3' UTRs indicated that the distinct length UTRs of the transcripts is attributed to an alternative usage of various polyadenylation signals in the 3' UTR. The open reading frame of 1203 bp encoded a putative 401 amino acid peptide. The deduced amino acid sequence shared 45-50% identities with the known B-type cyclin in other animals. Quantitative real-time RT-PCR revealed that the short transcript (1.7 kb) was the most abundant among the three transcripts, followed by the long (2.4 kb) and medium (1.9 kb), and the three forms of the transcripts displayed various expression profiles during oogenesis. In situ hybridization showed that the short transcript commenced expressing in the ova as early as the oogonia stage and accumulated largely at the perinucleolus (PN) stage, whereas almost no expression was found for the medium and long transcripts at the oogonia stage and moderate signals were detected at the PN stage. The differential expression of the three forms of transcripts suggested that various transcripts might perform different roles during oogenesis of the kuruma prawn. PMID- 15878300 TI - Lies, damn lies and statistics. PMID- 15878301 TI - Bone morphogenetic proteins in dentin regeneration for potential use in endodontic therapy. AB - The human dentition is indispensable for nutrition and physiology. The teeth have evolved for mastication of food. Caries is a common dental problem in which the dentin matrix is damaged. When the caries is deep and the dental pulp is exposed, the pulp has to be removed in many cases, resulting ultimately in loss of the tooth. Therefore, the regeneration of dentin-pulp complex is the long-term goal of operative dentistry and endodontics. The key elements of dentin regeneration are stem cells, morphogens such as bone morphogenetic proteins (BMPs) and a scaffold of extracellular matrix. The dental pulp has stem/progenitor cells that have the potential to differentiate into dentin-forming odontoblasts in response to BMPs. Pulpal wound healing consists of stem/progenitor cells release from dental pulp niche after noxious stimuli such as caries, migration to the injured site, proliferation and differentiation into odontoblasts. There are two main strategies for pulp therapy to regenerate dentin: (1) in vivo method of enhancing the natural healing potential of pulp tissue by application of BMP proteins or BMP genes, (2) ex vivo method of isolation of stem/progenitor cells, differentiation with BMP proteins or BMP genes and transplantation to the tooth. This review summarizes recent advances in application of BMPs for dentin regeneration and possible use in endodotic therapy. PMID- 15878302 TI - Tagged magnetic resonance imaging of the heart: a survey. AB - Magnetic resonance imaging (MRI) of the heart with magnetization tagging provides a potentially useful new way to assess cardiac mechanical function, through revealing the local motion of otherwise indistinguishable portions of the heart wall. While still an evolving area, tagged cardiac MRI is already able to provide novel quantitative information on cardiac function. Exploiting this potential requires developing tailored methods for both imaging and image analysis. In this paper, we review some of the progress that has been made in developing such methods for tagged cardiac MRI, as well as some of the ways these methods have been applied to the study of cardiac function. PMID- 15878303 TI - Mechanistic perspectives for 1,2,4-trioxanes in anti-cancer therapy. AB - In addition to their well-known anti-malarial activity, artemisinin and its derivatives (1,2,4-trioxanes) possess potent activity against tumor cells in the nano- to micromolar range. Candidate genes that may contribute to the sensitivity and resistance of tumor cells to artemisinins were identified by pharmacogenomic and molecular pharmacological approaches. Target validation was performed using cell lines transfected with candidate genes or corresponding knockout cells. These genes are from classes with different biological function; for example, regulation of proliferation (BUB3, cyclins, CDC25A), angiogenesis (vascular endothelial growth factor and its receptor, matrix metalloproteinase-9, angiostatin, thrombospondin-1) or apoptosis (BCL-2, BAX). Artesunate triggers apoptosis both by p53-dependent and -independent pathways. Anti-oxidant stress genes (thioredoxin, catalase, gamma-glutamyl-cysteine synthetase, glutathione S transferases) as well as the epidermal growth factor receptor confer resistance to artesunate. Cell lines over-expressing genes that confer resistance to established anti-tumor drugs (MDR1, MRP1, BCRP, dihydrofolate reductase, ribonucleotide reductase) were not cross-resistant to artesunate, indicating that this drug has a different target and is not subject to multidrug resistance. The Plasmodium translationally controlled tumor protein (TCTP) represents a known target protein of artemisinin and its derivatives in the malaria parasite. The microarray-based mRNA expression of human TCTP correlated with sensitivity to artesunate in tumor cells, suggesting that human TCTP contributes to response of tumor cells to the drug. The multi-factorial nature of cellular response to artemisinin and its derivatives may be beneficial to treat otherwise drug resistant tumors and may explain why resistance development has not been observed in either cancer or malaria. PMID- 15878304 TI - Cytogenetic studies in mouse oocytes irradiated in vitro at different stages of maturation, by use of an early preantral follicle culture system. AB - In vivo studies on X-irradiated mice have shown that structural chromosome aberrations can be induced in female germ cells and that the radiation-induced chromosomal damage strongly depends on the stage of maturation reached by the oocytes at the time of irradiation. In the present study, the sensitivity of oocytes to induction of chromosome damage by radiation was evaluated at two different stages, by use of a recently developed method of in vitro culture covering a crucial period of follicle/oocyte growth and maturation. A key feature of this system is that growth and development of all follicles is perfectly synchronized, due to the selection of a narrow class of follicles in the start off culture. This allows irradiation of well-characterized and homogenous populations of follicles, in contrast to the situation prevailing in vivo. Follicles were X-irradiated with either 2 or 4 Gy, on day 0 of culture (early preantral follicles with one to two cell layers) or on day 12, 3h after hormonal stimulation of ovulation (antral Graafian follicles). Ovulated oocytes, blocked in metaphase I (MI) by colchicine, were fixed 16 h after hormonal stimulation and analyzed for chromosome aberrations. The results confirm the high radiosensitivity of oocytes at 2 weeks prior to ovulation and the even higher radiosensitivity of those irradiated a few hours before ovulation, underlining the suitability of the in vitro system for further studies on the genetic effects of ionising radiation in female mammals. PMID- 15878305 TI - Effect of topiramate on cognitive function and activity level following neonatal seizures. AB - Topiramate, an antiepileptic drug with a number of mechanisms of action including blockade of AMPA/KA receptor subtypes, was assessed as a neuroprotective agent following seizures. We administered topiramate or saline chronically during and following a series of 25 neonatal seizures. After completion of the topiramate treatment, animals were tested in the water maze for spatial learning and the open field for activity level. Brains were then examined for cell loss and sprouting of mossy fibers. Rats treated with topiramate performed significantly better in the water maze than rats treated with saline. Topiramate treatment also reduced the amount of seizure-induced sprouting in the supragranular region. There were no differences between topiramate- and saline-treated rats in activity level in the open field, swimming speed, or weight gain. These findings show that long-term treatment with topiramate after neonatal seizures changes the long-term consequences of seizures and improves cognitive function. PMID- 15878306 TI - Comparison of classifications of seizures: a preliminary study with 28 participants and 48 seizures. AB - PURPOSE: Our aim was to compare three available seizure classifications (SCs), namely, the international classification of epileptic seizures published in 1981 (ICES; Epilepsia 1981;22:489-50); the semiological seizure classification (SSC) by H. Luders, J. Acharya, C. Baumgartner, et al. (Epilepsia 1998;39:1006-13; Acta Neurol Scand 1999;99:137-41); and the proposal of a new diagnostic scheme for seizures (PDSS) by J. Engel, Jr. (Epilepsia 2001;42:796-803) published in 2001. The three SCs were compared with respect to diagnostic success rates, usefulness, and consistency by a large group of neurologists in this preliminary study. METHODS: After a training period, 28 blindfed participants with different levels of experience with epilepsy classified videos or written descriptions of 48 randomly selected seizures according to the three SCs. Definite diagnoses of the seizures were established based on all clinical, ictal/interictal EEG, and MRI data. All the participants answered a questionnaire concerning their preferences for SCs after the study. RESULTS: The overall diagnostic success rates were 81.4% for ICES, 80.5% for PDSS, and 87.5%, for SSC. Various parameters concerning experience with epilepsy affected success rates positively, without reaching statistical significance, whereas experience with epilepsy surgery appeared to be a parameter significantly affecting the success rate in all SCs. In reliability analysis, Cronbach's alpha was 0.94 for ICES, 0.88 for PDSS, and 0.70 for SSC, all showing good agreement in the group. Nineteen reviewers chose SSC, eight chose ICES, and one chose PDSS as their preference in the questionnaire, completed after the end of the study. CONCLUSION: The results of this preliminary study demonstrate that with proper training, physicians treating epilepsy patients can handle new SCs, and emphasize the need for revision of the current classification. PMID- 15878307 TI - Common characteristics of epilepsy and schizophrenia: clinical observation and depth electrode studies. 1961. PMID- 15878308 TI - Atypical language representation in epilepsy: implications for injury-induced reorganization of brain function. AB - This review addresses language function and reorganization associated with various forms of epilepsy. Longstanding epilepsy, particularly types with onset early in life, may be associated with changes in the representation of language function in the brain. As a result of this reorganization, language function may be relatively spared despite injury to areas of the brain that normally subserve these functions. We examine the changes seen in language function in two types of epilepsy: hemispheric epilepsy of childhood and focal epilepsies. Findings from behavioral studies, intracarotid amytal testing, intraoperative cortical testing, and more recent functional imaging studies are reviewed. Studying changes in the representation of language function seen in some forms of epilepsy provides information about brain plasticity with implications for other neurologic diseases, as well as for the neuroscientific understanding of how and when functional reorganization may occur. PMID- 15878309 TI - New metallothionein mRNAs in Gobio gobio reveal at least three gene duplication events in cyprinid metallothionein evolution. AB - This paper reports the identification and analysis of the primary structure of three novel metallothionein cDNA sequences in the gudgeon, Gobio gobio (Cyprinidae). Two different 180 bp coding regions were identified, resulting in two MT isoforms differing in one amino acid. The primary structure of the amino acid sequence was compared to other cyprinid MT sequences. Furthermore, two differently sized cDNAs were discovered in one of the two transcripts. We present a phylogenetic comparison of our sequences to other, previously published cyprinid MT gene sequences. Our analysis reveals an unexpected complexity in cyprinid MT evolution, with at least three gene duplication events. Differences and homologies between the evolution of cyprinid MT genes are compared to other teleost families. Finally, possible implications for metallothionein classification are discussed. PMID- 15878310 TI - Overexpression of Rad51 inhibits double-strand break-induced homologous recombination but does not affect gene conversion tract lengths. AB - DNA double-strand breaks (DSBs) in yeast are repaired by homologous recombination (HR) and non-homologous end-joining (NHEJ). Rad51 forms nucleoprotein filaments at processed broken ends that effect strand exchange, forming heteroduplex DNA (hDNA) that gives rise to a gene conversion tract. We hypothesized that excess Rad51 would increase gene conversion tract lengths. We found that excess Rad51 reduced DSB-induced HR but did not alter tract lengths or other outcomes including rates of crossovers, break-induced replication, or chromosome loss. Thus, excess Rad51 appears to influence DSB-induced HR at an early stage. MAT heterozygosity largely mitigated the inhibitory effect of excess Rad51 on allelic HR, but not direct repeat HR. Excess Rad52 had no effect on DSB-induced HR efficiency or outcome, nor did it mitigate the dominant negative effects of excess Rad51. Excess Rad51 had little effect on DSB-induced lethality in wild type cells, but it did enhance lethality in yku70Delta mutants. Interestingly, dnl4Delta showed marked DSB-induced lethality but this was not further enhanced by excess Rad51. The differential effects of yku70Delta and dnl4Delta indicate that the enhanced killing with excess Rad51 in yku70Delta is not due to its NHEJ defect, but may reflect its defect in end-protection and/or its inability to escape from checkpoint arrest. Srs2 displaces Rad51 from nucleoprotein filaments in vitro, suggesting that excess Rad51 might antagonize Srs2. We show that excess Rad51 does not reduce survival of wild-type cells treated with methylmethane sulfonate (MMS), or cells suffering a single DSB. In contrast, excess Rad51 sensitized srs2Delta cells to both MMS and a single DSB. These results support the idea that excess Rad51 antagonizes Srs2, and underscores the importance of displacing Rad51 from nucleoprotein filaments to achieve optimum repair efficiency. PMID- 15878311 TI - Characterisation of the ventilatory response to hypoxia in a model of transgenic anemic mice. AB - Both polycythemia and the increase in hypoxic ventilatory response (HVR) are considered as important factors of acclimatization to hypoxia. The objective of this study was to characterise the ventilation pattern at different inspired oxygen fraction in a model of chronic anemic mice. These mice have a targeted disruption in the 5' untranslated region of the Epo gene that reduces Epo expression such that the homozygous animal is severely anemic. Ventilation in normoxia in Epo-TAg(h) mice was significantly greater than in wild type, and the difference was mainly due to a higher tidal volume. HVR was higher in Epo-TAg(h) mice at every FIO2 suggesting a higher chemosensitivity. Resting oxygen consumption was maintained in anemic mice. Maximal oxygen consumption was 30% lower while hemoglobin was 60% lower in anemic mice compared to wild type. This small decrease in maximal oxygen consumption is probably due a greater cardiac output and/or a better tissue oxygen extraction and would allow these anemic mice to acclimatize to hypoxia in spite of low oxygen carrying capacity. In conclusion, Epo-TAg(h) anemic mice showed increased ventilation and hypoxic ventilatory response. However, whether these adaptations will contribute to acclimatization in chronic hypoxia remains to be determined. PMID- 15878312 TI - Quantification of urinary o,o'-dityrosine, a biomarker for oxidative damage to proteins, by high performance liquid chromatography with triple quadrupole tandem mass spectrometry. A comparison with ion-trap tandem mass spectrometry. AB - We recently described an isotope dilution reversed-phase liquid chromatography atmospheric pressure chemical ionization-ion-trap-tandem mass spectrometry (HPLC APCI-MS/MS) method for the quantitative determination of oxidized amino acids in human urine, including o,o'-dityrosine, a specific marker of protein oxidation. In the present study, we investigated the possibility to use a triple quadrupole instrument for the analysis of this biomarker in urine. The two instruments were compared in terms of sensitivity, specificity and reproducibility. Results showed that the triple quadrupole instrument reaches 2.5-fold higher sensitivity (LOD=0.01 microM) compared to the previously used ion-trap instrument. Precision of the present assay is as follows: in-day variation is 4.6% and inter-day variation is 17%. The currently developed method was applied to a group of smoker urine samples. The mean urinary o,o'-dityrosine concentration was 0.08+/-0.01 microM. Expressed per urinary creatinine concentration, this corresponds to 10.1+/-0.4 micromol/mol creatinine. This is comparable to the previously reported values of 5.8+/-0.3 micromol/mol creatinine in non-smokers night-time urines, and 12.3+/-5 micromol/mol creatinine in day-time urines measured by the ion-trap instrument. PMID- 15878313 TI - Investigation of the separation of heterocyclic aromatic amines by reversed phase ion-pair liquid chromatography coupled with tandem mass spectrometry: the role of ion pair reagents on LC-MS/MS sensitivity. AB - Reversed phase ion-pair chromatography (RP-IPC) of seven heterocyclic aromatic amines encompassing quinoline (IQ, MeIQ), quinoxaline (MeIQx), pyridine (PhIP) and carboline derivatives (AalphaC, Harman, Norharman) was carried out with formate as counter ion in an aqueous eluent with acetonitrile as organic modifier. TSKgel ODS-80TS was used as the stationary phase. With the aim of acquiring a better insight into the mutual influence of ion-pair reagent and the organic modifier upon solute retention, the study was performed by using an experimental design approach able to evidencing the effect of the simultaneous variation of the two factors. A model for the chromatographic behavior of the amines is proposed that includes classical ion-pair mechanism involving formate in the case of MeIQx, PhIP, Harman and Norharman. A competitive ion-exchange mechanism was hypothesized to govern retention of quinoline compounds, whereas electrostatic interactions and hydrogen bond formation with the silanols of the stationary phase were judged to be responsible for the retention of AalphaC. Further, the chromatographic behavior of the analytes using the formic acid ammonium formate buffer in the mobile phase was compared with that observed using acetic acid-ammonium acetate buffer. The method based on the use of RP IPC with tandem mass spectrometry when the eluent contained formate buffer at pH 2.8 exhibited higher detectability with respect to that achieved using the acetate buffer. PMID- 15878314 TI - Osteopetrosis-like phenotype in latent TGF-beta binding protein 3 deficient mice. AB - LTBPs are extracellular matrix proteins resembling fibrillins. LTBP-1, 3, and 4 covalently bind latent TGF-beta and modulate tissue levels of this potent cytokine through regulation of its secretion, localization, and/or activation. To address LTBP function in vivo, we generated Ltbp-3 null mice. Ltbp-3-/- animals developed craniofacial abnormalities due to early ossification of the skull base synchondroses and displayed reduced body size. In addition, histological examination of Ltbp-3-/- skeletons revealed an increase in bone mass. The osteoblast numbers and mineral apposition rates were decreased in Ltbp-3-/- mice, whereas the osteoclast numbers were similar in null and wild type mice. Histological examination revealed persistence of cartilage remnants in Ltbp-3-/- trabecular bone. Taken together, these results indicate that the Ltbp-3-/- high bone mass phenotype was due to a defect in bone resorption. We hypothesize that lack of Ltbp-3 results in decreased levels of TGF-beta in bone and cartilage, which leads to compromised osteoclast function and decreased bone turnover. PMID- 15878315 TI - Endogenous expression and endocytosis of tartrate-resistant acid phosphatase (TRACP) by osteoblast-like cells. AB - Tartrate-resistant acid phosphatase (TRACP) is produced by macrophages and other cells of the monohistiocytic lineage. In particular, osteoclasts are characterized for a high expression of this enzyme. Yet, several data suggest that other bone cell types, such as osteocytes and osteoblasts, may also express activity of this enzyme. This is particularly obvious at sites were osteoclasts resorb bone, suggesting that osteoclasts (or their precursors) somehow induce TRACP activity in osteoblasts. In the present study, we investigated this by culturing human osteoblast-like cells with and without conditioned medium (MCM) from human blood monocytes (as a source of osteoclast precursors). High levels of TRACP activity were found in osteoblast-like cells cultured with MCM. Depletion of TRACP from this medium resulted in the absence of its activity in osteoblast like cells, thus suggesting that the TRACP activity in these cells was the result of endocytosed TRACP that was released by the monocytes in the MCM. Osteoblast like cells cultured in control (non-conditioned) medium contained very low levels of TRACP-like activity. However, the cells expressed TRACP mRNA and incubation of extracts of these cells with active cathepsin B did induce activity of a TRACP like enzyme. Inhibition of the activity of cysteine proteinases in general and of cathepsin B in particular, completely blocked TRACP activity of the osteoblast like cells. This TRACP-like enzyme but not the alleged endocytosed fraction of TRACP was inhibited by fluoride, suggesting that the fractions may be different isoenzymes. Our data seem to indicate that osteoblast-like cells may contain two different fractions of TRACP, one that is released by monocytes and subsequently endocytosed by osteoblast-like cells and a second endogenous fraction that is present in an inactive proform. We hypothesize that the capacity of osteoblast like cells to endocytose TRACP is important for the removal of this enzyme during or following the bone resorptive activity of the osteoclast. PMID- 15878316 TI - Loading induces site-specific increases in mineral content assessed by microcomputed tomography of the mouse tibia. AB - Adaptation to mechanical loading has been studied extensively in cortical, but not cancellous bone. However, corticocancellous sites are more relevant to osteoporosis and related fracture risk of the hip and spine. We tested the hypotheses that adaptation in a long bone would be greater at cancellous than cortical sites and would depend on the term of daily in vivo cyclic axial loading. We applied compressive loads to the adolescent, 10-week old, male C57BL/6 mouse tibia to examine the skeletal response immediately prior to attainment of peak bone mass. Adaptation was quantified at the completion of either 2-week (n = 8) or 6-week (n = 12) loading terms by directly comparing volumetric bone mineral content between loaded and contralateral limbs by microcomputed tomography. The increase in mineral content was site specific with a greater response found in the corticocancellous proximal metaphysis (14%) than the cortical mid-shaft (2%) after 6 weeks of loading. Furthermore, bone volume fraction and average trabecular thickness of cancellous bone in the proximal tibia increased after 6 weeks by 15% and 12% respectively. Diaphyseal response was only evident proximal to the mid-shaft as indicated by an 8% increase in maximum principal moment of inertia. Both loading terms produced similar results for mineral content, volume fraction, and moments of inertia. Our finding that non-invasive loading increases the bone volume and fraction at a corticocancellous site by as much as 15% motivates exploring the use of mechanical loading to attain greater peak bone mass and inhibit osteoporosis. PMID- 15878317 TI - Anabolic actions of PTH (1-34): use of a novel tissue engineering model to investigate temporal effects on bone. AB - PTH is in clinical use for the treatment of osteoporosis and is under intensive investigation for its potential in applications of tissue engineering, fracture healing, and implant integration. However, the mechanisms of its action to stimulate bone formation are still unclear. A novel bone tissue engineering model was used to elucidate basic mechanisms of PTH anabolic actions. Ectopic ossicles containing cortical bone, trabecular bone, and a hematopoietic marrow were generated from implanted bone marrow stromal cells (BMSC). One week after implantation, nude mice were administered PTH or vehicle for 1 week (group 1), 3 weeks (group 2), or 7 weeks (group 3). Another group was also treated for 3 weeks, initiated 12 weeks after implantation (group 4). Micro-radiography and histomorphometry revealed increased marrow cellularity in group 1 PTH-treated ossicles, increased bone in group 2 PTH-treated ossicles, and similar amounts of bone in both group 3 and 4 ossicles regardless of treatment. Incidence of phosphate mineral and phosphate mineral to hydroxyproline ratio via Raman spectroscopy were significantly higher after 3 weeks versus 1 week of PTH treatment, but there was no difference between PTH- and vehicle-treated ossicles. Early events of PTH action in group 1 ossicles and the effects of a single injection of PTH on 1- and 2-week-old ossicles were evaluated by Northern blot analysis. Osteocalcin (OC) mRNA was increased after 1 week of intermittent PTH treatment in ossicles and calvaria but an acute injection did not alter OC mRNA. In contrast, a single injection of PTH increased matrix gamma-carboxyglutamic acid protein (MGP) mRNA in 2-week-old ossicles. Differential and temporal dependent effects of PTH on OC and MGP suggest at the molecular level, that PTH acts to inhibit osteoblast mineralization. However, this does not translate into tissue level alterations. These data indicate that anabolic actions of PTH in ectopic ossicles are temporally dependent on the BMSC implanted and suggest that cell implantation strategies are particularly responsive to PTH. PMID- 15878318 TI - Effects of teriparatide [rhPTH (1-34)] treatment on structural geometry of the proximal femur in elderly osteoporotic women. AB - INTRODUCTION: We evaluated effects of teriparatide (rDNA origin) injection [teriparatide, rhPTH (1-34), TPTD] on hip structure among a subset 558 postmenopausal women enrolled in the Fracture Prevention Trial. METHODS: Patients were randomized to once-daily, self-administered subcutaneous injections of placebo (N = 189), teriparatide 20 mug (TPTD20; N = 186), or 40 mug (TPTD40; N = 183) for a median of 20 months. Repeated dual energy X-ray absorptiometry (DXA) hip scans were analyzed with the Hip Structure Analysis (HSA) program to derive structural geometry. RESULTS AND CONCLUSIONS: There were no significant differences in age or body size between groups at baseline, 1 year, or study termination. At the femoral neck, teriparatide increased bone mass and improved bone geometric strength in both treatment groups compared to the placebo group, with the response being dose-related. The mean difference (95% CI) in bone cross sectional area (CSA) in the TPTD20 was 3.5% (1.8% to 5.3%), and 6.3% (4.5% to 8.2%) in TPTD40 at study termination, compared to placebo controls. Teriparatide treatment increased bending strength, with the mean difference in section modulus being 3.6% (1.4% to 5.8%) and 6.8% (4.6% to 9.1%) greater in the TPTD20 and TPTD40 groups, respectively. Compared to placebo, local cortical instability characterized by the buckling ratio decreased by 5.5% (3.5% to 7.5%) and 8.6% (6.6% to 10.5%) in the TPTD20 and TPTD40 groups, respectively, during the study period. The changes at the intertrochanteric region were comparable to those at the narrow neck although between-group differences were slightly smaller. Except for an inconsequential (1%) improvement in section modulus in TPTD20, teriparatide effects did not reach significance at the femoral shaft. In conclusion, teriparatide treatment improved axial and bending strength, and increased cortical thickness and stability at the femoral neck and intertrochanteric region. Teriparatide treatment effects were not apparent at the purely cortical femoral shaft. PMID- 15878319 TI - Inhibition of angiogenesis and tumor growth by beta and gamma-secretase inhibitors. AB - The involvement of beta-secretase and gamma-secretase in producing the beta amyloid component of senile plaques found in the brain of Alzheimer's patients has fueled a major research effort to design selective inhibitors of these proteases. Interestingly, gamma-secretase cleaves several proteins including Notch, E-cadherin, CD44 and ErbB-4 (erythroblastic leukemia viral oncogene homolog 4), which are important modulators of angiogenesis. The beta-amyloid precursor protein, which is cleaved by beta-secretase and gamma-secretase to produce beta-amyloid, is highly expressed in the endothelium of neoforming vessels suggesting that it might play a role during angiogenesis. These data prompted us to explore the effects of beta and gamma-secretase inhibitors of different structures on angiogenesis and tumor growth. Both the gamma and beta secretase inhibitors tested reduce endothelial cell proliferation without inducing cellular toxicity, suppress the formation of capillary structures in vitro and oppose the sprouting of microvessel outgrowths in the rat aortic ring model of angiogenesis. Moreover, they potently inhibit the growth and vascularization of human glioblastoma and human lung adenocarcinoma tumors xenotransplanted into nude mice. Altogether these data suggest that the gamma and beta-secretases play an essential role during angiogenesis and that inhibitors of the beta and gamma-secretases may constitute new classes of anti-angiogenic and anti-tumoral compounds. PMID- 15878320 TI - Allosteric modulation of 5-HT3 serotonin receptors. AB - [(3)H]Granisetron binding to 5-HT(3) type serotonin receptors was examined in homogenates of rat forebrain and NG 108-15 cells. We have applied an allosteric model to 5-HT(3) receptor binding for the first time. Slope factors of displacement improved the modelling. Serotonin displaced [(3)H]granisetron binding with micromolar potency in forebrain and with nanomolar potency in NG 108 15 cells. Racemic and (+)verapamil, ifenprodil and GYKI-46903 were used as representative allosteric inhibitors of 5-HT(3) receptors. They displaced [(3)H]granisetron binding with great negative cooperativity (alpha>10) and exerted great negative cooperativity with serotonin binding (beta>10). Great negative cooperativity of these agents with serotonin and [(3)H]granisetron binding cannot be distinguished from dual competitive displacement. Trichloroethanol (data from literature) had no cooperativity with [(3)H]granisetron binding (alpha~1) and exhibit positive cooperativity with serotonin (beta<1) in displacement. The allosteric model can lead to a more quantitative method in vitro to develop allosteric agents for 5-HT(3) receptors. PMID- 15878321 TI - Anxiolytic-like effects of mGlu1 and mGlu5 receptor antagonists in rats. AB - The purpose of the present study was to compare anxiolytic activity of the metabotropic glutamate receptor 1 (mGlu) antagonist, EMQMCM ((3-ethyl-2-methyl quinolin-6-yl)-(4-methoxy-cyclohexyl)-methanone methanesulfonate) and the mGlu5 receptor antagonist MTEP ([(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine) and MPEP (2-methyl-6-(phenylethynyl)pyridine) in animal models of anxiety. In the elevated plus maze, diazepam (1 mg/kg), but not the mGlu1 or mGlu5 receptor antagonists induced anxiolytic-like effects. Meanwhile, MTEP (2.5 and 5 mg/kg), EMQMCM (5 mg/kg), and diazepam (2 mg/kg) all significantly inhibited fear potentiated startle. In the contextual fear conditioning test, MTEP (1.25 and 2.5 but not 5 mg/kg) and EMQMCM (0.6 to 5 mg/kg) attenuated freezing responding. In the Geller Seifter conflict test, MPEP (1 and 3 mg/kg), MTEP (3 mg/kg), chlordiazepoxide (10 and 20 mg/kg) and midazolam (1 mg/kg) all facilitated punished responding, while ECMQCM failed to produce any significant effects up to 3 mg/kg dose. To summarise, the present data further support a significant anxiolytic potential of group I mGlu receptor antagonists, while suggesting the effects of mGlu1 receptor antagonists may depend on the experimental procedure and may be qualitatively different from those of mGlu5 receptor antagonists. PMID- 15878322 TI - Different antithrombotic properties of factor Xa inhibitor and thrombin inhibitor in rat thrombosis models. AB - We compared the antithrombotic properties of a factor Xa inhibitor (DX-9065a) with those of a thrombin inhibitor (melagatran) in a rat disseminated intravascular coagulation model and a rat venous thrombosis model. Rat disseminated intravascular coagulation and venous thrombosis models were produced by injection of tissue factor and platinum wire placement, respectively. DX-9065a exerted antithrombotic effects dose dependently in both models. Melagatran was also effective in the venous thrombosis model, whereas it showed an aggravation in the disseminated intravascular coagulation model at low but not high doses. In the in vitro study, DX-9065a decreased the C(max) of the thrombin generation curve in plasma irrespective of whether protein C was present or not. However, melagatran increased the C(max) at low concentrations when protein C was present. This increase was not detected in protein C-deficient plasma. These results suggest that, unlike DX-9065a, melagatran in low doses aggravates disseminated intravascular coagulation by increasing thrombin generation, which may be partly due to suppression of negative feedback by activated protein C. PMID- 15878323 TI - Mechanism-based modeling of reduced inotropic responsiveness to digoxin in endotoxemic rat hearts. AB - The mechanisms by which endotoxemia affects myocardial contractility and responsiveness to inotropic drugs are not well understood. We examined the positive inotropic effect of digoxin in single-pass Langendorff-perfused hearts from rats after in vivo pretreatment with lipopolysaccharide (LPS, 4 mg/kg, i.p., 4 h before heart isolation). Using a mathematical modeling approach that allows differentiation between effects elicited at the receptor and postreceptor level, we studied uptake, receptor binding and effectuation kinetics after three consecutive digoxin doses (15, 30, and 45 microg) in the absence and presence of the reverse mode Na(+)/Ca(2+) exchange (NCX) inhibitor KB-R7943 (0.1 microM) in perfusate. LPS significantly depressed baseline contractility and the inotropic response to digoxin without affecting its uptake mechanism. Compared with the control group, the slope of the functional receptor occupancy (stimulus)-to response relationship was reduced by 44% in the LPS group. Model analysis revealed a significant correlation between changes in digoxin action and LPS induced febrile response: digoxin receptor affinity increased and the response/stimulus ratio decreased with rise in body temperature, respectively. In contrast, the diminished responsiveness to digoxin observed after NCX inhibition in the control group was not further attenuated in the LPS group. These results support the hypothesis that postreceptor events may be responsible for the diminished contractile response to digoxin during endotoxemia. PMID- 15878324 TI - Protective effect of astaxanthin on naproxen-induced gastric antral ulceration in rats. AB - Frequently used for humans as non-steroidal anti-inflammatory drug, naproxen has been known to induce ulcerative gastric lesion. The present study investigated the in vivo protective effect of astaxanthin isolated from Xanthophyllomyces dendrorhous against naproxen-induced gastric antral ulceration in rats. The oral administration of astaxanthin (1, 5, and 25 mg/kg of body weight) showed a significant protection against naproxen (80 mg/kg of body weight)-induced gastric antral ulcer and inhibited elevation of the lipid peroxide level in gastric mucosa. In addition, pretreatment of astaxanthin resulted in a significant increase in the activities of radical scavenging enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. A histologic examination clearly proved that the acute gastric mucosal lesion induced by naproxen nearly disappeared after the pretreatment of astaxanthin. These results suggest that astaxanthin removes the lipid peroxides and free radicals induced by naproxen, and it may offer potential remedy of gastric ulceration. PMID- 15878325 TI - Hyperalgesia in non-obese diabetic (NOD) mice: a role for the inducible bradykinin B1 receptor. AB - Most studies performed to investigate the role of the inducible bradykinin B(1) receptor in the pathology and complications of type 1 diabetes have been carried out using the model of streptozotocin (STZ)-induced diabetes. The model of spontaneous autoimmune diabetes in non-obese diabetic (NOD) mice involves a long term inflammatory process that closely resembles the human type 1 diabetes. In the present study, we aimed at establishing the correlation between the progress of diabetic hyperalgesia and the incidence of diabetes, as a function of age, in NOD mice. We also evaluated the implication of the bradykinin B(1) receptor, a receptor up-regulated during the inflammatory progress of diabetes, in the development of diabetic hyperalgesia in NOD mice. Female NOD mice were followed up from the 4th to the 32nd week of age for the incidence of diabetes. Only NOD mice with plasma glucose concentration >20 mmol/l were considered diabetic. The nociception was assessed using the hot plate and the tail immersion pain tests and the effect of acute and chronic administration of the selective bradykinin B(1) receptor agonist, desArg(9)bradykinin and its selective antagonists, R-715 (Ac-Lys-[D-beta Nal(7), Ile(8)]desArg(9)bradykinin) and R-954 (Ac-Orn-[Oic(2), alpha-MePhe(5), D-beta Nal(7), Ile(8)]desArg(9)bradykinin), on the development of diabetic hyperalgesia was studied. Diabetic NOD mice developed a significant time dependent hyperalgesia, as measured in both tests, starting from the 8th week of age with the maximum effect observed over 16 to 20 weeks, whereas the incidence of diabetes in the tested NOD mice was only 40.16% at the age of 16 weeks and reached a maximum of 73.23% at the age 24 weeks. Both acute and chronic administration of desArg(9)bradykinin (400 microg/kg) markedly increased the hyperalgesic activity in diabetic NOD mice in the hot plate and tail immersion nociceptive tests. The selective bradykinin B(1) receptor antagonist R-715 (400 microg/kg) and its more potent and long acting analogue R-954 (200 microg/kg), administered in acute or chronic manner, significantly attenuated diabetic hyperalgesia in NOD mice in both thermal pain tests and restored nociceptive responses to values observed in control non-diabetic siblings. Our results bring the first evidence that the development of hyperalgesia in NOD mice, a model of spontaneous type 1 diabetes, precedes the occurrence of hyperglycemia and is mediated by the bradykinin B(1) receptor. It is suggested that bradykinin B(1) receptor antagonism could become a novel therapeutic approach to the treatment of diabetic neuropathic complications. PMID- 15878326 TI - Effects of a selective bradykinin B1 receptor antagonist on increased plasma extravasation in streptozotocin-induced diabetic rats: distinct vasculopathic profile of major key organs. AB - Diffuse vasculopathy is a common feature of the morbidity and increased mortality associated with insulino-dependent type 1 diabetes. Increased vascular permeability leading to plasma extravasation occurs in surrounding tissues following endothelial dysfunction. Such micro- and macro-vascular complications develop over time and lead to oedema, hypertension, cardiomyopathy, renal failure (nephropathy) and other complications (neuropathy, retinopathy). In the present investigation, we studied the effect of a selective bradykinin B(1) receptor antagonist, R-954, on the enhanced vascular permeability in streptozotocin (STZ) induced diabetic Wistar rats compared with age-matched controls. Plasma extravasation was determined using Evans blue dye in selected target tissues (left and right heart atria, ventricles, lung, abdominal and thoracic aortas, liver, spleen, renal cortex and medulla), at 1 and 4 weeks following STZ administration. The vascular permeability was significantly increased in the aortas, cortex, medulla, and spleen in 1-week STZ rats and remained elevated at 4 weeks of diabetes. Both atria showed an increased vascular permeability only after 4-week STZ-administration. R-954 (2 mg/kg, bolus, s.c.), given 2 h prior to Evans blue dye, to 1- and 4-week diabetic rats significantly inhibited (by 48 100%) plasma leakage in most tested tissues affected by diabetes with no effect in healthy rats. These results showed that the inducible bradykinin B(1) receptor subtype participates in the modulation of the vascular permeability in diabetic rats and suggest that selective bradykinin B(1) receptor antagonism could have a beneficial role in reducing diabetic vascular complications. PMID- 15878327 TI - Reduction of apoptosis through the mitochondrial pathway by the administration of acetyl-L-carnitine to mouse fibroblasts in culture. AB - It is shown in literature that stress, such as deprivation of trophic factors and hypoxia, induces apoptosis in cultured cells and in tissues. In light of these results, we explored the possibility of protecting cells from programmed death by improving the metabolism of the mitochondrion. To this end, acetyl-L-carnitine was administered at various concentrations under conditions of serum deprivation. The choice of this drug was based on the accepted notion that acetyl-L-carnitine is able to stabilize mitochondrial membranes and to increase the supply of energy to the organelle. The results presented here indicate that the drug protects cells from apoptotic death: this is demonstrated by a lower positivity to the TUNEL reaction and by a strong reduction of the apoptotic DNA ladder in serum deprived cells. The involvement of the mitochondrial apoptotic pathway was assessed by cytochrome C release and immunoreactivity to caspase 3. Moreover, acetyl-L-carnitine stimulates cell proliferation. PMID- 15878328 TI - Identification of a novel cell-adhesive protein spatiotemporally expressed in the basement membrane of mouse developing hair follicle. AB - We used PCR-based cDNA subtraction to screen for genes up-regulated during mouse hair morphogenesis. One gene selected was predominantly expressed at the tip of developing hair follicles and encoded a protein characterized by the presence of twelve tandem repeats of approximately 120 amino acids and a novel N-terminal domain containing an Arg-Gly-Asp cell-adhesive motif. Immunohistochemistry demonstrated that the protein encoded by this gene, named QBRICK, was localized at the basement membrane zone of embryonic epidermis and hair follicles, in which it was more enriched at the tip rather than the stalk region. Cell adhesion assays showed that QBRICK was active in mediating cell-substratum adhesion through integrins containing alphav or alpha8 chain, but not integrin alpha5beta1. Immunohistochemistry showed that QBRICK colocalized with alphav containing integrins in the interfollicular region, but with the alpha8 containing integrin at the tip region of developing hair follicles. These results, together, indicate that QBRICK is an adhesive ligand of basement membrane distinctively recognized by cells in the embryonic skin and hair follicles through different types of integrins directed to the Arg-Gly-Asp motif. PMID- 15878329 TI - Characterization of the human GARP (Golgi associated retrograde protein) complex. AB - The Golgi associated retrograde protein complex (GARP) or Vps fifty-three (VFT) complex is part of cellular inter-compartmental transport systems. Here we report the identification of the VFT tethering factor complex and its interactions in mammalian cells. Subcellular fractionation shows that human Vps proteins are found in the smooth membrane/Golgi fraction but not in the cytosol. Immunostaining of human Vps proteins displays a vesicular distribution most concentrated at the perinuclear envelope. Co-staining experiments with endosomal markers imply an endosomal origin of these vesicles. Significant accumulation of VFT complex positive endosomes is found in the vicinity of the Trans Golgi Network area. This is in accordance with a putative role in Golgi associated transport processes. In Saccharomyces cerevisiae, GARP is the main effector of the small GTPase Ypt6p and interacts with the SNARE Tlg1p to facilitate membrane fusion. Accordingly, the human homologue of Ypt6p, Rab6, specifically binds hVps52. In human cells, the "orphan" SNARE Syntaxin 10 is the genuine binding partner of GARP mediated by hVps52. This reveals a previously unknown function of human Syntaxin 10 in membrane docking and fusion events at the Golgi. Taken together, GARP shows significant conservation between various species but diversification and specialization result in important differences in human cells. PMID- 15878330 TI - Function of donor cell centrosome in intraspecies and interspecies nuclear transfer embryos. AB - Centrosomes, the main microtubule-organizing centers (MTOCs) in most animal cells, are important for many cellular activities such as assembly of the mitotic spindle, establishment of cell polarity, and cell movement. In nuclear transfer (NT), MTOCs that are located at the poles of the meiotic spindle are removed from the recipient oocyte, while the centrosome of the donor cell is introduced. We used mouse MII oocytes as recipients, mouse fibroblasts, rat fibroblasts, or pig granulosa cells as donor cells to construct intraspecies and interspecies nuclear transfer embryos in order to observe centrosome dynamics and functions. Three antibodies against centrin, gamma-tubulin, and NuMA, respectively, were used to stain the centrosome. Centrin was not detected either at the poles of transient spindles or at the poles of first mitotic spindles. gamma-tubulin translocated into the two poles of the transient spindles, while no accumulated gamma-tubulin aggregates were detected in the area adjacent to the two pseudo-pronuclei. At first mitotic metaphase, gamma-tubulin was translocated to the spindle poles. The distribution of gamma-tubulin was similar in mouse intraspecies and rat-mouse interspecies embryos. The NuMA antibody that we used can recognize porcine but not murine NuMA protein, so it was used to trace the NuMA protein of donor cell in reconstructed embryos. In the pig-mouse interspecies reconstructed embryos, NuMA concentrated between the disarrayed chromosomes soon after activation and translocated to the transient spindle poles. NuMA then immigrated into pseudo pronuclei. After pseudo-pronuclear envelope breakdown, NuMA was located between the chromosomes and then translocated to the spindle poles of first mitotic metaphase. gamma-tubulin antibody microinjection resulted in spindle disorganization and retardation of the first cell division. NuMA antibody microinjection also resulted in spindle disorganization. Our findings indicate that (1) the donor cell centrosome, defined as pericentriolar material surrounding a pair of centrioles, is degraded in the 1-cell reconstituted embryos after activation; (2) components of donor cell centrosomes contribute to the formation of the transient spindle and normal functional mitotic spindle, although the contribution of centrosomal material stored in the recipient ooplasm is not excluded; and (3) components of donor cell centrosomes involved in spindle assembly may not be species-specific. PMID- 15878331 TI - Loss of the Dictyostelium RasC protein alters vegetative cell size, motility and endocytosis. AB - In addition to its previously established roles in cAMP relay and cAMP chemotaxis, loss of signal transduction through the RasC protein was found to impact a number of vegetative cell functions. Vegetative rasC- cells exhibited reduced random motility, were less polarized and had altered F-actin distribution. Cells lacking RasC also contained more protein and were larger in size than wild type cells. These increases were associated with increased liquid phase endocytosis. Despite the increase in cell size, cytokinesis was relatively normal and there was no change in the rate of cell division. rasC- cells also chemotaxed poorly to folate and exhibited reduced F-actin accumulation, reduced ERK2 phosphorylation and reduced Akt/PKB phosphorylation in response to folate, indicating that RasC was also involved in transducing chemotactic signals in vegetative cells. PMID- 15878332 TI - Vav promotes differentiation of human tumoral myeloid precursors. AB - Vav is one of the genetic markers that correlate with the differentiation of hematopoietic cells. In T and B cells, it appears crucial for both development and functions, while, in non-lymphoid hematopoietic cells, Vav seems not involved in cell maturation, but rather in the response of mature cells to agonist dependent proliferation and phagocytosis. We have previously demonstrated that the amount and the tyrosine phosphorylation of Vav are up-regulated in both whole cells and nuclei of tumoral promyelocytes induced to granulocytic maturation by ATRA and that tyrosine-phosphorylated Vav does not display any ATRA-induced GEF activity but contributes to the regulation of PI 3-K activity. In this study, we report that Vav accumulates in nuclei of ATRA-treated APL-derived cells and that the down-modulation of Vav prevents differentiation of tumoral promyelocytes, indicating that it is a key molecule in ATRA-dependent myeloid maturation. On the other hand, the overexpression of Vav induces an increased expression of surface markers of granulocytic differentiation without affecting the maturation-related changes of the nuclear morphology. Consistent with an effect of Vav on the transcriptional machinery, array profiling shows that the inhibition of the Syk dependent tyrosine phosphorylation of Vav reduces the number of ATRA-induced genes. Our data support the unprecedented notion that Vav plays crucial functions in the maturation process of myeloid cells, and suggest that Vav can be regarded as a potential target for the therapeutic treatment of myeloproliferative disorders. PMID- 15878333 TI - Characterization of the telomere complex, TERF1 and TERF2 genes in muntjac species with fusion karyotypes. AB - The telomere binding proteins TRF1 and TRF2 maintain and protect chromosome ends and confer karyotypic stability. Chromosome evolution in the genus Muntiacus is characterized by numerous tandem (end-to-end) fusions. To study TRF1 and TRF2 telomere binding proteins in Muntiacus species, we isolated and characterized the TERF1 and -2 genes from Indian muntjac (Muntiacus muntjak vaginalis; 2n = 6 female) and from Chinese muntjac (Muntiacus reveesi; 2n = 46). Expression analysis revealed that both genes are ubiquitously expressed and sequence analysis identified several transcript variants of both TERF genes. Control experiments disclosed a novel testis-specific splice variant of TERF1 in human testes. Amino acid sequence comparisons demonstrate that Muntiacus TRF1 and in particular TRF2 are highly conserved between muntjac and human. In vivo TRF2-GFP and immuno-staining studies in muntjac cell lines revealed telomeric TRF2 localization, while deletion of the DNA binding domain abrogated this localization, suggesting muntjac TRF2 represents a functional telomere protein. Finally, expression analysis of a set of telomere-related genes revealed their presence in muntjac fibroblasts and testis tissue, which suggests the presence of a conserved telomere complex in muntjacs. However, a deviation from the common theme was noted for the TERT gene, encoding the catalytic subunit of telomerase; TERT expression could not be detected in Indian or Chinese muntjac cDNA or genomic DNA using a series of conserved primers, while TRAP assay revealed functional telomerase in Chinese muntjac testis tissues. This suggests muntjacs may harbor a diverged telomerase sequence. PMID- 15878334 TI - Mitochondrial localization of cyclooxygenase-2 and calcium-independent phospholipase A2 in human cancer cells: implication in apoptosis resistance. AB - Cyclooxygenase-2 (COX-2) is inducible by myriad stimuli. The inducible COX-2 in primary cultured human cells has been reported to localize to nuclear envelope, endoplasmic reticulum, nucleus and caveolae. As COX-2 plays an important role in tumor growth, we were interested in its subcellular location in cancer cells. We examined COX-2 localization in several cancer cell lines by confocal microscopy. A majority of COX-2 was colocalized with heat shock protein 60, a mitochondrial protein, in colon cancer (HT-29, HCT-15 and DLD-1), breast cancer (MCF7), hepatocellular cancer (HepG2) and lung cancer cells (A549) with a similar distribution pattern. By contrast, COX-2 was not localized to mitochondria in human foreskin fibroblasts or endothelial cells. Immunoblot analysis of COX-2 in mitochondrial and cytosolic fractions confirmed localization of COX-2 to mitochondria in HT-29 and DLD-1 cells but not in fibroblasts. Calcium-independent phospholipase A2 was colocalized with heat shock protein 60 to mitochondria not only in cancer cells (HT-29 and DLD-1) but also in fibroblasts. HT-29 which expressed more abundant mitochondrial COX-2 than DLD-1 was highly resistant to arachidonic acid and H2O2-induced apoptosis whereas DLD-1 was less resistant and human fibroblasts were highly susceptible. Treatment of HT-29 cells with sulindac or SC-236, a selective COX-2 inhibitor, resulted in loss of resistance to apoptosis. These results suggest that mitochondrial COX-2 in cancer cells confer resistance to apoptosis by reducing the proapoptotic arachidonic acid. PMID- 15878335 TI - Disruption of Smad5 gene induces mitochondria-dependent apoptosis in cardiomyocytes. AB - Our previous studies have shown that SMAD5, an important intracellular mediator of transforming growth factor beta (TGF-beta) family, is required for normal development of the cardiovascular system in vivo. In the current study, we reported that the lack of the Smad5 gene resulted in apoptosis of cardiac myocytes in vivo. To further investigate the mechanism of the Smad5 gene in cardiomyocyte apoptosis, the embryonic stem (ES) cell differentiation system was employed. We found that the myotubes that differentiated from the homozygous Smad5ex6/ex6 mutant ES cells underwent collapse and degeneration during the late stages of in vitro differentiation, mimicking the in vivo observation. By electron microscopy, abnormal swollen mitochondria were observed in cardiomyocytes both from Smad5-deficient embryos and from ES-differentiated cells. There was also a significant reduction in mitochondrial membrane potential (Deltapsi m) and a leakage of cytochrome c from mitochondria into the cytosol of myocytes differentiated from Smad5 mutant ES cells. The expression of p53 and p21 was found to be elevated in the differentiated Smad5 mutant myocytes, and this was accompanied by an up-regulation in caspase 3 expression. These results suggest that the Smad5-mediated TGF-beta signals may protect cardiomyocytes from apoptosis by maintaining the integrity of the mitochondria, probably through suppression of p53 mediated pathways. PMID- 15878336 TI - Sensitization to UV-induced apoptosis by the histone deacetylase inhibitor trichostatin A (TSA). AB - UV-induced apoptosis is a protective mechanism that is primarily caused by DNA damage. Cyclobutane pyrimidine dimers (CPD) and 6-4 photoproducts are the main DNA adducts triggered by UV radiation. Because the formation of DNA lesions in the chromatin is modulated by the structure of the nucleosomes, we postulated that modification of chromatin compaction could affect the formation of the lesions and consequently apoptosis. To verify this possibility we treated human colon carcinoma RKO cells with the histone deacetylase inhibitor trichostatin A (TSA) prior to exposure to UV radiation. Our data show that pre-treatment with TSA increased UV killing efficiency by more than threefold. This effect correlated with increased formation of CPDs and consequently apoptosis. On the other hand, TSA treatment after UV exposure rather than before had no more effect than UV radiation alone. This suggests that a primed (opened) chromatin status is required to sensitize the cells. Moreover, TSA sensitization to UV-induced apoptosis is p53 dependent. p53 and acetylation of the core histones may thus contribute to UV-induced apoptosis by modulating the formation of DNA lesions on chromatin. PMID- 15878337 TI - Carboxypeptidases cathepsins X and B display distinct protein profile in human cells and tissues. AB - Cathepsin X, a recently discovered lysosomal cysteine protease, shares common structural features and activity properties with cysteine protease cathepsin B. Based on its widespread mRNA distribution in primary tumors and tumor cell lines, a redundant function in tumor progression has been proposed. In this study, we have shown that these two related proteases exhibit different profiles with respect to their protein distribution in cells and tissues and to their possible roles in malignancy. Protein level of cathepsin X did not differ significantly between matched pairs of lung tumor and adjacent lung tissue obtained from patients with lung cancer whereas that of cathepsin B was 9.6-fold higher in tumor compared to adjacent lung tissue. Immunohistochemical analysis of lung tumor cathepsin X revealed very faint staining in tumor cells but positive staining in infiltrated histiocytes, alveolar macrophages, bronchial epithelial cells, and alveolar type II cells. Cathepsin X stained positive also in CD68+ cells in germinal centers of secondary follicles in lymph nodes, corresponding to tingible body macrophages. Two cell lines with proven invasive behavior, MCF-10A neoT and MDA-MB 231, showed positive staining for cathepsin B, but negative for cathepsin X. We showed that the invasive potential of MCF-10A neoT cells can be impaired by specific inhibitor of cathepsin B but not by that of cathepsin X. Cathepsin X was found in large amounts in the pro-monocytic U-937 cell line, in monocytes and in dendritic cells, generated from monocytes in vitro. Our results show that cathepsin X is not involved in degradation of extracellular matrix, a proteolytic event leading to tumor cell invasion and metastasis. Its expression, restricted to immune cells suggests a role in phagocytosis and the regulation of immune response. PMID- 15878338 TI - The multidomain protooncogenic protein c-Cbl binds to tubulin and stabilizes microtubules. AB - The protooncogenic protein c-Cbl is known to regulate the actin cytoskeleton. In this study, we present results indicating that c-Cbl can also regulate the microtubular network. We have shown that c-Cbl binds to tubulin and microtubules through its tyrosine kinase binding (TKB) domain. However, the character of the interactions described in this report is novel, since the G306E mutation, which disrupts the ability of c-Cbl's TKB to bind to tyrosine-phosphorylated proteins, does not affect the observed interaction between c-Cbl and microtubules. Furthermore, overexpression of c-Cbl in human pulmonary artery endothelial cells and COS-7 cells leads to microtubule stabilization. We demonstrate that this effect of c-Cbl is mediated by TKB, and, like c-Cbl binding to microtubules, is independent of the ability of TKB to bind to tyrosine-phosphorylated proteins. Finally, we have shown that c-Cbl directly polymerizes microtubules in vitro, and that TKB is necessary and sufficient for this effect of c-Cbl. In this last phenomenon, as well as in the previous ones, the effect of TKB is not sensitive to the inactivating G306E mutation. Overall, the results presented in this report suggest a novel function for c-Cbl-microtubule binding and stabilization. PMID- 15878339 TI - The immunomodulatory protein B7-H4 is overexpressed in breast and ovarian cancers and promotes epithelial cell transformation. AB - B7-H4 protein is expressed on the surface of a variety of immune cells and functions as a negative regulator of T cell responses. We independently identified B7-H4 (DD-O110) through a genomic effort to discover genes upregulated in tumors and here we describe a new functional role for B7-H4 protein in cancer. We show that B7-H4 mRNA and protein are overexpressed in human serous ovarian cancers and breast cancers with relatively little or no expression in normal tissues. B7-H4 protein is extensively glycosylated and displayed on the surface of tumor cells and we provide the first demonstration of a direct role for B7-H4 in promoting malignant transformation of epithelial cells. Overexpression of B7 H4 in a human ovarian cancer cell line with little endogenous B7-H4 expression increased tumor formation in SCID mice. Whereas overexpression of B7-H4 protected epithelial cells from anoikis, siRNA-mediated knockdown of B7-H4 mRNA and protein expression in a breast cancer cell line increased caspase activity and apoptosis. The restricted normal tissue distribution of B7-H4, its overexpression in a majority of breast and ovarian cancers and functional activity in transformation validate this cell surface protein as a new target for therapeutic intervention. A therapeutic antibody strategy aimed at B7-H4 could offer an exciting opportunity to inhibit the growth and progression of human ovarian and breast cancers. PMID- 15878340 TI - Possible role of deep tubular invaginations of the plasma membrane in MHC-I trafficking. AB - Tubules and vesicles are membrane carriers involved in traffic along the endocytic and secretory routes. The small GTPase Arf6 regulates a recycling branch of short dynamic tubular intermediates used by major histocompatibility class I (MHC-I) molecules to traffic through vesicles between endosomes and the plasma membrane. We observed that Arf6 also affects a second network of very long and stable tubules containing MHC-I, many of which correspond to deep invaginations of the plasma membrane. Treatment with wortmannin, an inhibitor of phosphatidylinositol-3-phosphate kinase, prevents formation of the short dynamic tubules while increasing the number of the long and very stable ones. Expression of NefAAAA, a mutant form of HIV Nef, increases the number of cells containing the stable tubules, and is used here as a tool to facilitate their study. Photoactivation of NefAAAA-PA-GFP demonstrates that this molecule traffics from endosomes to the tubules. Finally, live-cell imaging also shows internalization of MHC-I molecules into these tubules, suggesting that this is an additional route for MHC-I traffic. PMID- 15878341 TI - High levels of endogenous nitric oxide produced after burn injury in rats arrest activated T lymphocytes in the first G1 phase of the cell cycle and then induce their apoptosis. AB - Major physical traumas provoke a systemic inflammatory response and immune dysfunction. In a model of thermal injury in rats, we previously showed that an overproduction of nitric oxide (NO) was responsible for the collapse of lymphoproliferative responses. In the present work, we performed a time-course analysis of cell proliferation and cell death parameters in order to establish the sequence of events triggered by the high NO output in Wistar/Han rat splenocytes activated with Con A, 10 days after burn injury. We demonstrate that activated T cells from burned rats never divided whereas normal T cells underwent four division cycles. However, T cells from both burned and normal rat entered the G1 phase as shown by increase of cell size, mitochondria hyperpolarization, and expression of cyclin D1. Burned rat T cells progressed to the late G1 phase as shown by expression of the nuclear Ki-67 antigen, but they never entered the S phase. They underwent apoptosis as shown by morphological parameters, disruption of transmembrane mitochondrial potential, and DNA fragmentation. Persistent accumulation of the p53 protein accompanied these phenomena. NO synthase inhibitors antagonize alterations of cell proliferation and cell death parameters in burned rat T cells and accelerated p53 turnover. PMID- 15878342 TI - The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions. AB - Rsu-1 is a highly conserved leucine rich repeat (LRR) protein that is expressed ubiquitously in mammalian cells. Rsu-1 was identified based on its ability to inhibit transformation by Ras, and previous studies demonstrated that ectopic expression of Rsu-1 inhibited anchorage-independent growth of Ras-transformed cells and human tumor cell lines. Using GAL4-based yeast two-hybrid screening, the LIM domain protein, PINCH1, was identified as the binding partner of Rsu-1. PINCH1 is an adaptor protein that localizes to focal adhesions and it has been implicated in the regulation of adhesion functions. Subdomain mapping in yeast revealed that Rsu-1 binds to the LIM 5 domain of PINCH1, a region not previously identified as a specific binding domain for any other protein. Additional testing demonstrated that PINCH2, which is highly homologous to PINCH1, except in the LIM 5 domain, does not interact with Rsu-1. Glutathione transferase fusion protein binding studies determined that the LRR region of Rsu-1 interacts with PINCH1. Transient expression studies using epitope-tagged Rsu-1 and PINCH1 revealed that Rsu-1 co-immunoprecipitated with PINCH1 and colocalized with vinculin at sites of focal adhesions in mammalian cells. In addition, endogenous P33 Rsu-1 from 293T cells co-immunoprecipitated with transiently expressed myc-tagged PINCH1. Furthermore, RNAi-induced reduction in Rsu-1 RNA and protein inhibited cell attachment, and while previous studies demonstrated that ectopic expression of Rsu-1 inhibited Jun kinase activation, the depletion of Rsu-1 resulted in activation of Jun and p38 stress kinases. These studies demonstrate that Rsu-1 interacts with PINCH1 in mammalian cells and functions, in part, by altering cell adhesion. PMID- 15878343 TI - Activation of hypoxia-induced transcription in normoxia. AB - Hypoxia-inducible factor-1 (HIF-1), the master regulator of transcriptional responses to reduced oxygen tension (hypoxia) in mammalian cells, consists of one HIF-1alpha and one HIF-1beta subunit. In normoxia, HIF-1alpha subunits are hydroxylated on specific proline residues; modifications that signal ubiquitination and degradation of HIF-1alpha by the proteasome. To test the effect of saturating HIF-1alpha degradation, we generated a construct, denoted the saturating domain (SD), based on a region surrounding proline 564 (Pro564) in HIF-1alpha. Expression of the SD led to accumulation of endogenous HIF-1alpha proteins in nuclei of normoxic cells. The induced HIF-1alpha was functional as it activated expression from a hypoxia-regulated reporter gene and from the endogenous vascular endothelial growth facor-a (Vegf-a) and carbonic anhydrase 9 (Ca9) genes. The effect of the SD was dependent on Pro564 since a mutated SD, in which Pro564 had been replaced by a glycine residue, failed to bind the von Hippel-Lindau protein (pVHL) and to stabilise HIF-1alpha. Treatment of cells with the prolylhydroxylase inhibitor dimethyloxalylglycine, or the proteasome inhibitor MG-132, mimicked the effect of the SD. In conclusion, we show that blocking HIF-1alpha degradation, either by saturation, or inhibition of prolyl hydroxylases or proteosomal degradation, leads to nuclear localisation of active HIF-1alpha proteins. PMID- 15878344 TI - FHOD1 coordinates actin filament and microtubule alignment to mediate cell elongation. AB - Diaphanous-related formins (DRFs) are actin nucleators that mediate rearrangements of the actin cytoskeleton downstream of specific Rho GTPases. The DRF Formin Homology 2 Domain containing 1 (FHOD1) interacts with the Rac1 GTPase and induces the formation of and associates with bundled actin stress fibers. Here we report that active FHOD1 also coordinates microtubules with these actin stress fibers. Expression of a constitutive active FHOD1 variant in HeLa cells not only resulted in pronounced formation of FHOD1-actin fibers but also caused marked cell elongation and parallel alignment of microtubules without affecting cytokinesis of these cells. The analysis of deletions in the FH1 and FH2 functional regions revealed that the integrity of both domains was strictly required for FHOD1's effects on the cytoskeleton. Dominant-negative approaches demonstrated that filament coordination and cell elongation depended on the activity of the Rho-ROCK cascade, but did not involve Rac or Cdc42 activity. Experimental depolymerization of actin filaments or microtubules revealed that the formation of FHOD1-actin fibers was a prerequisite for the polarization of microtubules. However, only simultaneous disruption of both filament systems reversed the cell elongation induced by activated FHOD1. Thus, sustained cell elongation was a consequence of FHOD1-mediated actin-microtubule coordination. These results suggest filament coordination as a conserved function of mammalian DRFs. PMID- 15878345 TI - alpha-Crystallin localizes to the leading edges of migrating lens epithelial cells. AB - alpha-crystallin (alphaA and alphaB) is a major lens protein, which belongs to the small heat-shock family of proteins and binds to various cytoskeletal proteins including actin, vimentin and desmin. In this study, we investigated the cellular localization of alphaA and alphaB-crystallins in migrating epithelial cells isolated from porcine lens. Immunofluorescence localization and confocal imaging of alphaB-crystallin in confluent and in migrating subconfluent cell cultures revealed a distinct pattern of subcellular distribution. While alphaB crystallin localization was predominantly cytoplasmic in confluent cultures, it was strongly localized to the leading edges of cell membrane or the lamellipodia in migrating cells. In accordance with this pattern, we found abundant levels of alphaB-crystallin in membrane fractions compared to cytosolic and nuclear fractions in migrating lens epithelial cells. alphaA-crystallin, which has 60% sequence identity to alphaB-crystallin, also exhibited a distribution profile localizing to the leading edge of the cell membrane in migrating lens epithelial cells. Localization of alphaB-crystallin to the lamellipodia appears to be dependent on phosphorylation of residue serine-59. An inhibitor of p38 MAP kinase (SB202190), but not the ERK kinase inhibitor PD98059, was found to diminish localization of alphaB-crystallin to the lamellipodia, and this effect was found to be associated with reduced levels of Serine-59 phosphorylated alphaB crystallin in SB202190-treated migrating lens epithelial cells. alphaB-crystallin localization to the lamellipodia was also altered by the treatment with RGD (Arg Ala-Asp) peptide, dominant negative N17 Rac1 GTPase, cytochalasin D and Src kinase inhibitor (PP2), but not by the Rho kinase inhibitor Y-27632 or the myosin II inhibitor, blebbistatin. Additionally, in migrating lens epithelial cells, alphaB-crystallin exhibited a clear co-localization with the actin meshwork, beta catenin, WAVE-1, a promoter of actin nucleation, Abi-2, a component of WAVE-1 protein complex and Arp3, a protein of the actin nucleation complex, suggesting potential interactions between alphaB-crystallin and regulatory proteins involved in actin dynamics and cell adhesion. This is the first report demonstrating specific localization of alphaA and alphaB-crystallins to the lamellipodia in migrating lens epithelial cells and our findings indicate a potential role for alpha-crystallin in actin dynamics during cell migration. PMID- 15878346 TI - Autocrine regulation of human urothelial cell proliferation and migration during regenerative responses in vitro. AB - Regeneration of the urothelium is rapid and effective in order to maintain a barrier to urine following tissue injury. Whereas normal human urothelial (NHU) cells are mitotically quiescent and G0 arrested in situ, they rapidly enter the cell cycle upon seeding in primary culture and show reversible growth arrest at confluency. We have used this as a model to investigate the role of EGF receptor signaling in urothelial regeneration and wound-healing. Transcripts for HER-1, HER-2, and HER-3 were expressed by quiescent human urothelium in situ. Expression of HER-1 was upregulated in proliferating cultures, whereas HER-2 and HER-3 were more associated with a growth-arrested phenotype. NHU cells could be propagated in the absence of exogenous EGF, but autocrine signaling through HER-1 via the MAPK and PI3-kinase pathways was essential for proliferation and migration during urothelial wound repair. HB-EGF was expressed by urothelium in situ and HB-EGF, epiregulin, TGF-alpha, and amphiregulin were expressed by proliferating NHU cells. Urothelial wound repair in vitro was attenuated by neutralizing antibodies against HER-1 ligands, particularly amphiregulin. By contrast, the same ligands applied exogenously promoted migration, but inhibited proliferation, implying that HER-1 ligands provoke differential effects in NHU cells depending upon whether they are presented as soluble or juxtacrine ligands. We conclude that proliferation and migration during wound healing in NHU cells are mediated through an EGFR autocrine signalling loop and our results implicate amphiregulin as a key mediator. PMID- 15878347 TI - Cloning, characterization, and heat stress-induced redistribution of a protein homologous to human hsp27 in the zebrafish Danio rerio. AB - Hsp27 is a small heat shock protein (shsp) regulating stress tolerance and increasingly thought to play roles in tissue homeostasis and differentiation. The zebrafish Danio rerio is an important model for the study of developmental processes, but little is known regarding shsps in this animal. Here, we report the sequence, expression, regulation, and function of a zebrafish protein (zfHsp27) homologous to human Hsp27. zfHsp27 contains three conserved phosphorylatable serines and a cysteine important for regulation of apoptosis, but it lacks much of a C-terminal tail domain and shows low homology in two putative actin interacting domains that are features of mammalian Hsp27. zfHsp27 mRNA is most abundant in adult skeletal muscle and heart and is upregulated during early embryogenesis. zfHsp27 expressed in mammalian fibroblasts was phosphorylated in response to heat stress and anisomycin, and this phosphorylation was prevented by treatment with SB202190, an inhibitor of p38 MAPK. Expression of zfHsp27 and human Hsp27 in mammalian fibroblasts promoted a similar degree of tolerance to heat stress. zfHsp27 fusion proteins entered the nucleus and associated with the cytoskeleton of heat stressed cells in vitro and in zebrafish embryos. These results reveal conservation in regulation and function of mammalian and teleost Hsp27 proteins and define zebrafish as a new model for the study of Hsp27 function. PMID- 15878348 TI - A temperature-sensitive mutation in the WD repeat-containing protein Smu1 is related to maintenance of chromosome integrity. AB - Temperature-sensitive CHO-K1 mutant cell line tsTM18 exhibits chromosomal instability and cell cycle arrest at S and G2 phases with decreased DNA synthesis at the nonpermissive temperature, 39 degrees C. To identify the causative mutation, we fused tsTM18 cells with normal human cells to generate hybrids carrying fragments of human chromosomes. Analysis of chromosome content of temperature-resistant transformants and introduction of a bacterial artificial chromosome containing part of human chromosome 9 led to isolation of the human SMU1 gene. Comparison of sequences of the Smu1 gene from wild-type and mutant cells revealed that the mutant phenotype is caused by a G-to-A transition that yields a gly-to-arg substitution at position 489 in hamster Smu1. The substituted glycine is located in the WD-repeat domain of Smu1. Single-stranded DNA accumulated in the nuclei of mutant cells at 39 degrees C. Furthermore, cdc2 kinase was not activated during G2 phase, and there was no chromosome segregation due to incomplete assembly of the spindle during M phase. Thus, Smu1 appears to be involved directly or indirectly in DNA replication, activation of cdc2 kinase, spindle assembly, and maintenance of chromosome integrity, reflecting the important roles of Smu1 in cellular function. PMID- 15878349 TI - Oncostatin M induces an acute phase response but does not modulate the growth or maturation-status of liver progenitor (oval) cells in culture. AB - Following acute injury, the liver regenerates through hepatocyte division. If this pathway is impaired, liver repair depends on the recruitment of adult liver progenitor (oval) cells. Mice fed a choline deficient, ethionine supplemented (CDE) diet possess substantial numbers of oval cells, which can be isolated, or examined in vivo. Oncostatin M (OSM) has been shown to induce maturation of murine fetal hepatoblasts into hepatocytes. We recently confirmed this in human fetal liver cultures. Here, we show that liver OSM expression increases in mice fed a CDE diet and CDE-derived oval cell isolates express OSM and its receptor (OSMR). Oval cell lines (PIL cells), as well as primary oval cell cultures, displayed STAT-3 phosphorylation following OSM stimulation. OSM had no effect on the growth of primary oval cells, but it was pro-apoptotic to PIL cells, suggesting that the two cell models are not directly comparable. Expression of PCNA and cyclin D1 was not affected by OSM treatment. No evidence was obtained to suggest an effect on oval cell maturation with OSM treatment. However, decreased albumin production, accompanied by increased expression of haptoglobin and fibrinogen, suggests that OSM induced an acute phase reaction in cultured oval cells. PMID- 15878350 TI - Protein kinase C regulates the phosphorylation and oligomerization of ERM binding phosphoprotein 50. AB - Ezrin-Radixin-Moesin (ERM) binding phosphoprotein 50 (EBP50, a.k.a. NHERF-1) is a scaffold protein essential for the localization and coordinated activity of apical transporters, enzymes and receptors in epithelial cells. EBP50 acts via multiple protein binding interactions, including oligomerization through interactions of its PSD95-Dlg-ZO1 (PDZ) domains. EBP50 can be phosphorylated on multiple sites and phosphorylation of specific sites modulates the extent of oligomerization. The aim of the present study was to test the capacity of protein kinase C (PKC) to phosphorylate EBP50 and to regulate its oligomerization. In vitro experiments showed that the catalytic subunit of PKC directly phosphorylates EBP50. In HEK-293 cells transfected with rat EBP50 cDNA, a treatment with 12 myristate 13-acetate (PMA) induced a translocation of PKCalpha and beta isoforms to the membrane and increased 32P incorporation into EBP50. In co-transfection/co-precipitation studies, PMA treatment stimulated EBP50 oligomerization. Mass spectrometry analysis of full-length EBP50 and phosphorylation analyses of specific domains, and of mutated or truncated forms of EBP50, indicated that PKC-induced phosphorylation of EBP50 occurred on the Ser337/Ser338 residue within the carboxyl-tail domain of the protein. Truncation of Ser337/Ser338 also diminished PKC-induced oligomerization of EBP50. These results suggest the PKC signaling pathway can impact EBP50-dependent cellular functions by regulating EBP50 oligomerization. PMID- 15878351 TI - Nitric oxide reverses desferrioxamine- and hypoxia-evoked HIF-1alpha accumulation -implications for prolyl hydroxylase activity and iron. AB - Hypoxia inducible factor 1 (HIF-1) senses and coordinates cellular responses towards hypoxia. HIF-1 activity is primarily determined by stability regulation of its alpha subunit that is degraded by the 26S proteasome under normoxia due to hydroxylation by prolyl hydroxylases (PHDs) but is stabilized under hypoxia. Besides hypoxia, nitric oxide (NO) stabilizes HIF-1alpha and promotes hypoxia responsive target gene expression under normoxia. However, in hypoxia, NO attenuates HIF-1alpha stabilization and gene activation. It was our intention to explain the contrasting behavior of NO under hypoxia. We used the iron chelator desferrioxamine (DFX) or hypoxia to accumulate HIF-1alpha in HEK293 cells. Once the protein accumulated, we supplied NO donors and followed HIF-1alpha disappearance. NO-evoked HIF-1alpha destabilization was reversed by proteasomal inhibition or by blocking PHD activity. By using the von Hippel Lindau (pVHL)-HIF 1alpha capture assay, we went on to demonstrate binding of pVHL to HIF-1alpha under DFX/NO but not DFX alone. Showing increased intracellular free iron under conditions of hypoxia/NO compared to hypoxia alone, we assume that increased free iron contributes to regain PHD activity. Variables that allow efficient PHD activation such as oxygen availability, iron content, or cofactor accessibility at that end allow NO to modulate HIF-1alpha accumulation. PMID- 15878352 TI - Differential growth factor induction and modulation of human gastric epithelial regeneration. AB - While several autocrine/paracrine growth factors (GFs) can all stimulate epithelial regeneration in experimentally wounded primary gastric cultures, clinical relevance for their non-redundant cooperative actions in human gastric ulcer healing is suggested by the sequential pattern of GF gene induction in vivo. Using new HGE cell lines able to form a coherent monolayer with tight junctions as well as using primary human gastric epithelial cultures, we show that EGF, TGFalpha, HGF and IGFs accelerate epithelial restitution upon wounding, independently of the TGFbeta pathway (as opposed to intestinal cells). However, they differently modulate cell behavior: TGFalpha exerts strong effects (even more than EGF) on cytoplasmic spreading and non-oriented protruding activity of bordering cells whereas HGF preferentially coordinates single lamella formation, cell elongation and migration into the wound. IGF-I and IGF-II rather induce the alignment of bordering cells and maintain a compact monolayer front. The number of mitotic cells maximally increases with EGF, followed by TGFalpha and IGF-I, II. The current study demonstrates that GFs differentially regulate the regeneration of human gastric epithelial cells through specific modulation of cell shape adaptation, migration and proliferation, further stressing that a coordination of GF activities would be necessary for the normal progression of post-wounding epithelial repair. PMID- 15878353 TI - Marked increase in the histamine content of neointima after stent implantation of pig coronary artery and growth-promoting effects of histamine in cultured smooth muscle cells. AB - After coronary stent implantation, the unfavorable in-stent restenosis often occurs by the formation of neointima due to the proliferation of smooth muscle cells. Platelet-derived growth factor (PDGF) and other peptide growth factors contribute to this process, but little is known about the role of non-peptide factors in this process. In the present study, the role of histamine, a non peptide factor, in the formation of neointima was investigated using a pig coronary model of in-stent restenosis and a culture system of coronary smooth muscle cells. A Palmaz-Schatz stent was implanted in the left anterior descending coronary artery of male pigs. At 1, 2 and 4 weeks after stenting, the histamine content of neointima was determined to be 326 +/- 82, 1427 +/- 280 and 440 +/- 69 pmol/mg protein, respectively, by HPLC fluorometry. In contrast, the histamine content of arterial media from the untreated control arteries was only 15.3 +/- 1.6 pmol/mg protein. These results demonstrate that the histamine content of neointima is about 20 to 90-fold that of the normal media. In vitro, histamine by itself did not stimulate the proliferation of cultured smooth muscle cells, but potentiated the PDGF-stimulated proliferation of the cultured cells via a mechanism independent of H1 and H2 histamine receptors. Thus, histamine may be an important non-peptide factor in the pathogenesis of in-stent restenosis. PMID- 15878354 TI - Ganoderic acid X, a lanostanoid triterpene, inhibits topoisomerases and induces apoptosis of cancer cells. AB - Lanostanoid triterpenes isolated from Ganoderma amboinense were found to inhibit the growth of numerous cancer cell lines, and some of them inhibited the activities of topoisomerases I and IIalpha in vitro. Among the bioactive isolates, one of the most potent triterpene was identified to be 3 alpha-hydroxy 15 alpha-acetoxy-lanosta-7,9(11),24-trien-26-oic acid, ganoderic acid X (GAX). Treatment of human hepatoma HuH-7 cells with GAX caused immediate inhibition of DNA synthesis as well as activation of ERK and JNK mitogen-activated protein kinases, and cell apoptosis. Molecular events of apoptosis including degradation of chromosomal DNA, decrease in the level of Bcl-xL, the disruption of mitochondrial membrane, cytosolic release of cytochrome c and activation of caspase-3 were elucidated. The ability of GAX to inhibit topoisomerases and to sensitize the cancer cells toward apoptosis fulfills the feature of a potential anticancer drug. PMID- 15878355 TI - Hepatoprotective effects of Hibiscus, Rosmarinus and Salvia on azathioprine induced toxicity in rats. AB - As an anti-metabolite, Azathioprine inhibits the de novo and salvage pathways of purine synthesis. Intraperitoneal injection of this drug results in not only lymphocyte suppression but also toxicity to bone marrow, gastrointestinal tract, and liver. This Azathioprine-induced hepatotoxicity was found to be associated with oxidative damage. Plants with antioxidative properties have been traditionally used to prevent diseases associated with free radicals. In this report, we used water extracts of three herbal plants that have been commonly used for treating many illnesses (Hibiscus sabdariffa, Rosmarinus officinalis and Salvia officinalis). Here we show their novel hepatoprotective effects against Azathioprine-induced hepatotoxicity in rats. Typically, administration of Azathioprine induces oxidative stress through depleting the activities of antioxidants and elevating the level of malonialdehyde in liver. This escalates levels of alanine aminotransferase, and aspartate aminotranferase in serum. Pretreatment with any of the three herbal plants used in this investigation proved to have a protective effect against Azathioprine-induced hepatotoxicity. Animals pretreated with water extracts from any of the three herbs under investigation not only failed to show necrosis of the liver after azathioprine administration, but also retained livers that, for the most part, were histologically normal. In addition, these herbs blocked the induced elevated levels of alanine aminotransferase and aspartate aminotranferase in serum. The Azathioprine-induced oxidative stress was relieved to varying degrees by the examined herbal extracts. This effect was evident through reducing malonialdehyde levels and releasing the inhibitory effect of Azathioprine on the activities of glutathione, catalase and superoxide dismutase. To our knowledge, this report is the first that shows hepatoprotective effects of Hibiscus, Rosmarinus and Salvia species against Azathioprine-induced acute liver damage. PMID- 15878356 TI - Isoliquiritigenin induces apoptosis and cell cycle arrest through p53-dependent pathway in Hep G2 cells. AB - Isoliquiritigenin (ISL) is a natural pigment with the simple chalcone structure 4,2',4'-trihydroxychalcone. In this study, we report ISL induced inhibition in the proliferation of human hepatoma cells (Hep G2) for the first time. The cell proliferation inhibition achieved by ISL treatment resulted in a G2/M-phase arrest and programmed cell death. ISL treatment was found to result in the upregulation of p53, p21/WAF1, Fas/APO-1 receptor, Fas ligand, Bax and NOXA, but not in Bad. To elevate the role of p53 in these functions, we generated Hep G2 cells that express the dominant negative p53, which blocks the transcriptional activity of p53. The enhancement of p21/WAF1, Fas/APO-1, Bax and NOXA were decreased in Hep G2 cells that lack functional p53. Furthermore, Hep G2 cells were significantly more resistant to ISL when the activity of p53 was blocked. These results demonstrated that ISL-inducible p53 plays a key apoptotic role, and may do so by regulating the expression of specific target molecules that promotes efficient apoptotic cell death following G2/M-cell cycle arrest. PMID- 15878357 TI - Artelastin is a cytotoxic prenylated flavone that disturbs microtubules and interferes with DNA replication in MCF-7 human breast cancer cells. AB - Artelastin, a novel prenylated flavone, previously isolated from the wood bark of Artocarpus elasticus, was evaluated for its capacity to inhibit the growth of fifty-two human tumor cell lines, representing nine different tumor types. A pronounced dose-dependent growth inhibitory effect was detected in all the cell lines, with GI50 values ranging from 0.8-20.8 microM. Studies to elucidate the basis of the growth inhibitory activity of artelastin were performed in the MCF-7 human breast cancer cell line (GI50 = 6.0 microM). We show that artelastin exerts a biphasic effect in the DNA synthesis of MCF-7 cells, a stimulatory effect at low concentrations (below GI50) for short times of exposition (6 h and 24 h), and an inhibitory effect at high concentrations (above GI50). Remarkably, treated cells that have DNA synthesis affected could be viable and metabolically active. Furthermore, artelastin acts as a cytotoxic rather than a cytostatic compound. Massive cytoplasmatic vacuoles were detected in cells after artelastin treatment. Together with these morphological alterations, cells show the presence of abnormal nuclear morphologies, and occasionally nuclear condensation, which were identified as apoptotic by TUNEL assay. Moreover, artelastin was shown to disturb the microtubule network while no effect was observed on the kinetochores. Flow cytometry analysis of cells treated with artelastin reveal an accumulation in S phase that interferes with the cell cycle progression. Additionally, according to BrdU patterns, studies with synchronized cells at G0 and at G1/S transition also suggest that artelastin delays DNA replication since progression of cells trough S-phase is perturbed. PMID- 15878358 TI - Cardioprotection without cardiosuppression by SEA0400, a novel inhibitor of Na+ Ca2+ exchanger, during ischemia and reperfusion in guinea-pig myocardium. AB - The effect of SEA0400, a novel Na+-Ca2+ exchanger inhibitor, on mechanical and electrophysiological parameters of coronary-perfused guinea-pig right ventricular tissue preparation was examined during no-flow ischemia and reperfusion. Contractile force and action potential duration were decreased during no-flow ischemia, while the resting tension was increased. Upon reperfusion, transient arrhythmias were observed and contractile force returned to less than 50% of preischemic values. SEA0400 (1 microM) had no effect on the decline in contractile force during the no-flow ischemia, but abolished the rise in resting tension. SEA0400 significantly improved the recovery of contractile force after reperfusion to about 80% of the preischemic value. SEA0400 had no effect on the action potential under normal conditions and during ischemia, but significantly improved the recovery of action potential duration after reperfusion. Enhancement of the recovery of contractile force during reperfusion by SEA0400 was also observed when the drug was applied only before and during the ischemic period and when the drug was applied only during reperfusion. The present results indicate that inhibition of Na+-Ca2+ exchanger either during ischemia or during reperfusion exerts cardioprotective effects and enhances the recovery of myocardial contractile function. PMID- 15878359 TI - Neutrophils activation can be diminished by apolipoprotein A-I. AB - High density lipoprotein (HDL) has anti-inflammatory function. To investigate the effects of apolipoprotein A-I (ApoA-I), the major apolipoprotein of HDL, on activated neutrophils, we stimulated neutrophils in vitro with fMLP and PMA, as a receptor-binding and a nonreceptor-binding stimuli, respectively, and incubated ApoA-I with those neutrophils. Three conditions were utilized: 1) resting neutrophils + ApoA-I (0, 2.5,5, 10 microg/mL respectively), 2) fMLP(10(-7) mol/L) activated neutrophils + ApoA-I (0, 2.5, 5, 10 microg/mL respectively), and 3) PMA(10(-7) mol/L)-activated neutrophils + ApoA-I (0, 2.5, 5, 10 microg/mL respectively). After incubation, we measured neutrophils adhesion to fibronectin, oxidative bust (O2- and H2O2 production), degranulation (release of MPO and elastase), and L929 cell mortality which were attacked by release-out of cytokines in activated neutrophils (using MTT). Our results showed that in vitro ApoA-I inhibits fMLP- and PMA- activated neutrophil adhesion, oxidative burst, degranulation and L929 cell mortality. These inhibition effects of ApoA-I on fMLP activated neutrophils are more powerful than that on PMA-activated neutrophils. ApoA-I has no effect on resting neutrophils. We concluded that ApoA-I could diminish the function of activated neutrophils. PMID- 15878360 TI - 2-O-methyl PAF as a Ca2+ mobilizer in Madin Darby canine kidney cells. AB - In Madin-Darby canine kidney (MDCK) cells, the effect of 2-O-methyl PAF, an inactive analogue of platelet activating factor (PAF), on intracellular Ca2+ concentration ([Ca2+]i) was measured by using the Ca2+-sensitive fluorescent dye fura-2. 2-O-methyl PAF (> or = 15 microM) caused a rapid rise of [Ca2+]i in a concentration-dependent manner. 2-O-methyl PAF-induced [Ca2+]i rise was partly reduced by removal of extracellular Ca2+. 2-O-methyl PAF-induced extracellular Ca2+ influx was also suggested by Mn2+ influx-induced fura-2 fluorescence quench. The 2-O-methyl PAF-induced Ca2+ influx was blocked by nifedipine, verapamil and diltiazem. In Ca2+-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+-ATPase, caused a monophasic [Ca2+]i rise, after which 2-O-methyl PAF failed to increase [Ca2+]i; also, pretreatment with 2-O-methyl PAF depleted thapsigargin-sensitive Ca2+ stores. U73122, an inhibitor of phospholipase C, abolished ATP (but not 2-O-methyl PAF)-induced [Ca2+]i rise. These findings suggest that 2-O-methyl PAF evokes a rapid increase in [Ca2+]i in renal tubular cells by stimulating both extracellular Ca2+ influx and intracellular Ca2+ release. PMID- 15878361 TI - Ilex paraguariensis extracts are potent inhibitors of nitrosative stress: a comparative study with green tea and wines using a protein nitration model and mammalian cell cytotoxicity. AB - Due to the increasing importance of nitrosative stress in pathology and the efficacy displayed by flavonoids in cancelling the effects of peroxynitrite, we decided to conduct a comparative study of three commonly used beverages with the highest polyphenol contents and proven antioxidant properties: mate (Ilex paraguariensis); green tea (Camelia sinensis) extracts and white and red wines of the main varietals. We directly evaluated and compared the extracts and wines as protein nitration inhibitors using 3-nitrotyrosine as a biomarker, we studied the extracts as protectors from OONO-induced cytotoxicity in two mammalian cell lines. Both green tea and mate extracts have a high polyphenol content, in the case of Ip, its higher concentration and higher free radical quenching activity on the DPPH assay may be mainly due to the sui generis extraction procedure. When BSA was incubated in the presence of SIN-1, a time and dose dependent nitration of the protein is clearly shown. Co-incubation of BSA with Ip, green tea or red wines led to a dose dependent inhibition of the effect. Ip displayed the highest inhibitory activity, followed by red wines and the green tea. Dilutions as low as 1/1500 produced more than 80% inhibition of albumin nitration. When we studied peroxynitrite-induced cytotoxicity in murine RAW 264.7 macrophages and 31EG4 mammary cells., we found a potent, dose-dependent protective effect that was Ilex paraguariensis > red wines > green tea. Taken together, our results indicate that when the herbal preparations studied here are prepared the way they are usually drunk, Ip displays the highest inhibition of protein nitration, and the highest promotion of cell survival, whereas green tea or red wines display significant but lesser effects at the same concentrations. Further studies aiming at isolation of the active principles and assessment of their bioavailability are warranted. PMID- 15878362 TI - Effect of cobalt and chromium ions on MMP-1, TIMP-1, and TNF-alpha gene expression in human U937 macrophages: a role for tyrosine kinases. AB - Previous reports have suggested that the imbalance of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) activity may contribute to prosthetic loosening. However, the mechanisms controlling these enzymes in the periprosthetic environment is unknown. We examined the effect of Co2+ and Cr3+ ions on the expression of genes encoding MMP-1, one of the principal proteinases capable of degrading native fibrillar collagens in the extracellular matrix (ECM), its inhibitor TIMP-1, and TNF-alpha, a cytokine that plays a central role in the induction of implant osteolysis. Human U937 macrophages were incubated in suspension or on phosphorylcholine (PC)-polymer coated surfaces for 24h with Co2+ and Cr3+ ions. The level of mRNAs was determined by reverse transcription-polymerase chain reaction (RT-PCR). Results show that both Co2+ and Cr3+ ions induce the expression of MMP-1, TIMP-1, and TNF alpha mRNA in a dose-dependent manner in cell suspensions. Tyrosine kinase inhibitors have different effects on these stimulatory effects. Indeed, genistein has only partial inhibitory effect on MMP-1 and TIMP-1, with even less effect on TNF-alpha expression. In contrast, herbimycin A completely blocks MMP-1 and TNF alpha while partially inhibiting TIMP-1. However, Co2+ and Cr3+ ions had no effect on the expression of MMP-1 and TIMP-1 in macrophages cultured on the PC polymer, suggesting that the attachment of U937 macrophages to the PC-polymer surfaces may modify their gene expression. In fact, MMP-1 and TIMP-1 seems to be constitutively up-regulated in this condition. However, the effect of Co2+ and Cr3+ ions on macrophages cultured on PC-polymer coated surfaces is similar to what was observed in suspension. Together, these findings indicate that activation of MMP-1, TIMP-1, and TNF-alpha by Co2+ and Cr3+ ions is regulated by tyrosine kinases. PMID- 15878363 TI - Effect of vitamin E addition to poly(D,L)-lactic acid on surface properties and osteoblast behaviour. AB - Vitamin E (Vit.E, alpha-tocoferol) is a natural agent with anti-oxidative and anti-inflammatory properties and it has been suggested that it could act as a stimulating factor for osteoblast proliferation and maturation. We produced poly(D,L)-lactic acid films enriched with Vit.E (1, 5 and 10% w/w) and investigated their surface properties using the FTIR analysis, sessile measure of wettability and serum protein adsorption, and evaluated attachment and spreading of MC-3T3 E1 murine osteoblast cells. FTIR analysis showed the presence of Vit.E on the polymer surface and Vit.E increased the polymer wettability in a concentration-dependent manner. The serum total protein adsorption increased significantly onto the 10% Vit.E P(D,L)-LA and the main protein adsorbed was albumin. The presence of albumin, considered as an anti-adhesive protein, on the surface of Vit.E enriched P(D,L)-LA films (especially 5 and 10% Vit.E) could explain, at least in part, the behaviour of MC-3T3 osteoblast cells seeded onto the polymers. Cell adhesion and spreading were strongly decreased by Vit.E (5 and 10%) in spite of the increased wettability. This reaction could be cell type specific, independent by the surface wettability and linked to cell-specific characteristics (membrane phospholipid composition, integrins expression). Moreover a direct effect of Vit.E on cell adhesion and spreading cannot be completely excluded. PMID- 15878364 TI - Bone formation and resorption of highly purified beta-tricalcium phosphate in the rat femoral condyle. AB - The aim of this study was to examine the chronological histology associated with highly purified beta-tricalcium phosphate (beta-TCP) implanted in the rat femoral condyle. Specimens were harvested on days 4, 7, 14, 28 and 56 after implantation, and were analyzed by tartrate-resistant acid phosphatase (TRAP) staining, immunohistochemistry of the ED1 protein as a marker of the phagocyte system, and in situ hybridization with digoxigenin-labeled alpha1 chain of type I procollagen (COL1A1), osteopontin and osteocalcin. beta-TCP was resorbed in a chronological manner. Although new bone was not observed on day 4, fibroblast-like cells around beta-TCP were positive for COL1A1 and osteopontin mRNA. New bone formation presented after day 7. In the double-staining for OPN and ED1 on day 7, most cells around beta-TCP were positive for either osteopontin mRNA or ED1 protein. However, there were some doubly positive multinucleated cells, suggesting that they belonged to the mononuclear phagocyte system. After day 28, the implanted region was replaced with bone marrow. Multinucleated TRAP-positive and ED1 positive cells which adhered to beta-TCP at all stages seemed to be osteoclasts and they continuously resorbed beta-TCP. beta-TCP has a good biocompatibility since both bioresorption and bone formation started at an early stage after implantation. PMID- 15878365 TI - Developing macroporous bicontinuous materials as scaffolds for tissue engineering. AB - Calcareous skeletal elements (ossicles) isolated from the seastar, Pisaster giganteus, were characterized and tested as potential biocompatible substrates for cellular attachment. These ossicles have a remarkably robust open-framework architecture with an interconnected network of ca. 10 microm diameter pores. Scanning electron and confocal microscopy was used to characterize the cell substrate interaction. Cell culturing experiments revealed that the cells firmly attach to the ossicle surface, forming cell aggregates of several layers thick. The anchored cells extended to form 'bridges' between the openings in the bicontinuous framework and the degree of coverage increased as culture time progressed. Osteoblasts grown on the ossicles were found to be viable up to 32 days after initial seeding, as proven by assaying with AlamarBlue and FDA/PI staining indicating the ossicle's potential as an alternative highly effective tissue scaffold. Given the limitation in availability of this natural material, the results presented here should be seen as offering guidelines for future development of synthetic materials with physical and chemical properties strongly conducive to bone repair and restoration. PMID- 15878366 TI - Preparation of gamma-PGA/chitosan composite tissue engineering matrices. AB - Gamma-poly(glutamic acid) (gamma-PGA), a hydrophilic and biodegradable polymer, was chosen to modify chitosan matrices to produce a gamma-PGA/chitosan composite biomaterial. Three types of both dense and porous composite matrices containing different amounts of gamma-PGA were fabricated. Chitosan and gamma-PGA matrices were also prepared as controls. Fluorescence staining indicated that chitosan and gamma-PGA were evenly distributed in the composite matrices. SEM micrographs showed that an interconnected porous structure with a pore size of 30-100 microm was present in all porous matrices except the gamma-PGA ones. By increasing the percentage of gamma-PGA from 0% to 20%, the swelling ratio of the matrices was enhanced from 1.6 to 3.2. Similarly, the contact angle of the matrices decreased from 113 degrees to 94 degrees . These data suggested that the surface hydrophilicity, water absorption rate, and swelling ratio were improved by adding gamma-PGA to the matrices. Additionally, the mechanical strength of the porous gamma-PGA/chitosan matrices was about 25-50%, higher than that of the unmodified chitosan matrices. The composite matrices were also examined and found to be an appropriate environment for cell attachment and proliferation. The cell density on the 20% gamma-PGA-modified matrices was almost triple that on the unmodified chitosan matrices on day 5. In summary, the gamma-PGA/chitosan composite matrices, due to their better hydrophilic, cytocompatible, and mechanical properties, are very promising biomaterials for tissue engineering applications. PMID- 15878367 TI - A synthetic nanofibrillar matrix promotes in vivo-like organization and morphogenesis for cells in culture. AB - The purpose of this study was to design a synthetic nanofibrillar matrix that more accurately models the porosity and fibrillar geometry of cell attachment surfaces in tissues. The synthetic nanofibrillar matrices are composed of nanofibers prepared by electrospinning a polymer solution of polyamide onto glass coverslips. Scanning electron and atomic force microscopy showed that the nanofibers were organized into fibrillar networks reminiscent of the architecture of basement membrane, a structurally compact form of the extracellular matrix (ECM). NIH 3T3 fibroblasts and normal rat kidney (NRK) cells, when grown on nanofibers in the presence of serum, displayed the morphology and characteristics of their counterparts in vivo. Breast epithelial cells underwent morphogenesis to form multicellular spheroids containing lumens. Hence the synthetic nanofibrillar matrix described herein provides a physically and chemically stable three dimensional surface for ex vivo growth of cells. Nanofiber-based synthetic matrices could have considerable value for applications in tissue engineering, cell-based therapies, and studies of cell/tissue function and pathology. PMID- 15878368 TI - Concentrated hydroxyapatite inks for direct-write assembly of 3-D periodic scaffolds. AB - Hydroxyapatite (HA) scaffolds with a 3-D periodic architecture and multiscale porosity have been fabricated by direct-write assembly. Concentrated HA inks with tailored viscoelastic properties were developed to enable the construction of complex 3-D architectures comprised of self-supporting cylindrical rods in a layer-by-layer patterning sequence. By controlling their lattice constant and sintering conditions, 3-D periodic HA scaffolds were produced with a bimodal pore size distribution. Mercury intrusion porosimetry (MIP) was used to determine the characteristic pore size and volume associated with the interconnected pore channels between HA rods and the finer pores within the partially sintered HA rods. PMID- 15878369 TI - Subcutaneous xenotransplantation of bovine pancreatic islets. AB - Transplantation of pancreatic islets in diabetes is currently limited by the need of immunosuppressive therapy. The present study was designed to test an immunoprotection planar device for subcutaneous xenotransplantation of pancreatic islets in the diabetic rat. We tested three different devices made of polyethersulfone hollow fibers. In all diabetic rats, implantation of islet containing devices promptly normalized hyperglycemia. In vitro membrane permeability to glucose was correlated with implant function duration. These data confirm that bovine islets contained within devices and implanted subcutaneously remain functional for several days. Strategies to prolong islet function may allow achieving successful long-term islet implantation in this attractive site. PMID- 15878370 TI - Bioactivation of an anorganic bone matrix by P-15 peptide for the promotion of early bone formation. AB - This animal experiment compared the regenerative processes within defined bony defects of the porcine skull after delivery of routinely utilized bone graft materials: anorganic bone matrix (ABM) and an identical ABM carrying the cell binding peptide P-15. Particulated autogenous bone was used as a control group. The chosen porcine model guaranteed the transferability of the obtained results to clinical practice. A total observation period of 6 months was defined. The bone samples were examined microradiographically and histologically at 8 specific times. Sufficient osseointegration and osseoconduction could be demonstrated for both anorganic bone minerals. However, in the selected model significantly higher mineralization rates (p = 0.0286) were found in the microradiographic image at 12 weeks after application of the bioactive form. The histological examination confirmed this accelerating effect on bone formation starting at day 3. At the end of the study after 6 months, the mineralization values had equalized in both study groups. For the first time, the material was demonstrated to be suitable as a bone substitute material for the treatment of larger bony defects in a large animal model. The P-15 sequence accelerated the process of bone formation on the surface of the anorganic bone matrix as early as 3 days but was not traced over the whole term of the study. PMID- 15878371 TI - Effect of passage number and matrix characteristics on differentiation of endothelial cells cultured for tissue engineering. AB - Cells can sense the physical and chemical properties of artificial materials used as scaffolds for tissue engineering and regulate their behavior. Therefore, biomimetic and biospecific molecules are coated on materials to regulate function of cells on the tissue-engineered product. These bioactive molecules can be attached in a defined spectrum, concentration and spatial distribution in order to control adhesion, growth, viability, differentiation, and function of the cells. When autologous cells are used for tissue engineering, initially limited cells obtained may often need an amplification of cell number by passage in tissue culture before they are seeded on a biomaterial or scaffold. We have conducted this study to understand how the characteristics of bioactive molecule coating might affect proliferation, apoptosis and differentiation when endothelial cell (EC) is serially passaged. Proliferation was assessed by proliferating cell nuclear antigen (PCNA) staining along with counting of cells harvested from confluent monolayer. Apoptosis was assessed by Annexin V staining and differentiation by semi quantitative reverse transcriptase polymerase chain reaction (RT-PCR) for von Willebrand factor (vWF) expression and quantification of its release using enzyme linked immunosorbant assay (ELISA), and thrombogenicity by comparing platelet adhesion to EC monolayer Dacron grafts (DG) with specific protein coating. The results indicate that ECs easily lose its proliferation potential when they are cultured repeatedly on gelatin, turn apoptotic and over express the prothrombotic protein- vWF. Whereas, when it is grown on a matrix composed of fibrin, fibronectin, gelatin and vascular EC growth factor (VEGF), the cells retained their ability to proliferate, remained viable and were relatively less thrombogenic, even when passage number progressed. It is concluded that if ECs are grown on the composite matrix that mimics natural vessel scaffold, the cell number can be amplified without affecting its normal physiological function and may be used to generate effective tissue-engineered cardiovascular constructs. PMID- 15878372 TI - Down regulation of degenerative cartilage molecules in chondrocytes grown on a hyaluronan-based scaffold. AB - Hyaluronic-acid-based biomaterials used for cartilage repair allow the expression of specific extracellular matrix molecules by human chondrocytes grown onto them. We investigated whether these biomaterials could also create an environment in which the cells downregulate the expression of some catabolic factors. Chondrocytes were isolated from human articular cartilage obtained from the knees of patients with a history of trauma. First, the cells were expanded in monolayers and then they were seeded on a hyaluronic-acid derivative scaffold. Constructs and surnatants were collected and analysed at 1, 3, 7, 14 and 21 days after seeding. Immunohistochemical analysis for CD44 and caspase was carried out on paraffin-embedded sections. The Tunel method was used to identify chondrocyte apoptosis status. Secretion of MMP-1 and MMP-13 in the surnatants of the cells grown onto the biomaterial was measured by enzyme-linked immunosorbent assay. Nitric oxide (NO) production was evaluated by estimating the stable NO metabolite nitrite by the Griess method. A real-time RT-PCR analysis was performed on the constructs to evaluate the expression of type I and II collagens, aggrecan, Sox 9, MMP-1 and MMP-13 mRNAs at the different experimental times evaluated. Decreased levels of metalloproteinases and nitric oxide were observed in the surnatants of chondrocytes grown onto the hyaluronan-based scaffold. This was also confirmed by real-time PCR analysis which showed that the cells expressed the specific differentiated phenotype downregulating the expression of some catabolic molecules. Cells apoptosis decreased during the culture period, which further supported the biochemical data. The ability of the hyaluronan scaffold to reduce the expression and production of molecules involved in cartilage degenerative diseases indicates its use to treat early lesions of osteoarthritic patients. PMID- 15878373 TI - Cellular and molecular events during chondrogenesis of human mesenchymal stromal cells grown in a three-dimensional hyaluronan based scaffold. AB - Mesenchymal stromal cells (MSCs) seem to be a good alternative to chondrocytes for cartilage regeneration. To obtain new information on the sequence of cellular and molecular events during in vitro chondrogenic differentiation we analysed MSCs on a widely used hyaluronic acid biomaterial (Hyaff-11). Cellular differentiation was induced using two different concentrations of TGFbeta1 (10 and 20 ng/ml) and the process was analysed at different time points (24 h, and 7, 14, 21 and 28 days) using techniques of light and electron microscopy, real-time PCR and immunohistochemistry. We found that without TGFbeta MSCs did not survive while in the presence of TGFbeta the cells significantly proliferated from day 7 until day 28. Light and electron microscopy showed that TGFbeta at 20 ng/ml better induced the formation of cartilage-like tissue. Real-time PCR showed an increased expression of collagen type II, IX and aggrecan associated to a down regulation of collagen type I. Immunohistochemical analysis confirmed that collagen type I was down-modulated while collagen type II increased from day 14 to day 28. These data clearly showed that higher concentrations of TGFbeta (20 ng/ml) induce chondrogenesis of MSCs on Hyaff-11 scaffold better than 10 ng/ml of TGFbeta. This process is characterized by a sequence of cellular and molecular events that deal with the in vitro formation of a cartilage-like tissue. PMID- 15878374 TI - Effect of bone mineral density and amorphous diamond coatings on insertion torque of bone screws. AB - In this study, the potential of high-quality amorphous diamond (AD) coatings in reducing the torque and failures of bone screws was studied. Torque values were recorded for 32 stainless steel screws, 2.7 or 3.5 mm in diameter and 60 mm in length. Half of the screw sets were coated with the AD coating before installing in predrilled holes of human cadaveric femoral bone samples. The bone samples were selected from two groups of four persons with mean ages of 34 years (range 25-41 years) and 75 years (range 73-77 years), respectively. The bone mineral density (BMD) values of the samples were determined exactly at the screw insertion site by peripheral quantitative computed tomography (pQCT). In the mechanical tests, insertion and removal torques were measured. BMD had a significant effect on insertion torque; the maximum torque (adjusted with respect to the screw diameter) was significantly higher for the young bone than for the old bone (p < 0.05). By using a polished AD coating, insertion torque was decreased even up to 50% in comparison with the screws without coating. The results suggest that AD coating provides a stable, smooth surface and reduces the risk of screw failures. PMID- 15878375 TI - Fe-based nanoparticulate metallic alloys as contrast agents for magnetic resonance imaging. AB - Pharmaceutical grade magnetic colloidal dispersions have been prepared from iron alloys synthesized by laser pyrolysis. The colloids were obtained by simultaneous dispersion and coating of the particles with dextran in a strong alkaline solution. Both powders and dispersions have been analyzed in terms of microstructural characteristics, chemical composition and magnetic properties. The powders consist of uniform spherical nanoparticles (12 nm of diameter) showing a metallic core encapsulated into an iron-oxide shell. On the other hand, the colloidal dispersions consist of magnetic particles-aggregates with hydrodynamic sizes of approximately 75 nm. Magnetic resonance images of rats were taken after the intravenously administration of the Fe colloidal dispersions, and compared with those obtained using a commercial iron oxide magnetic resonance imaging contrast agent. The results showed a contrast improvement of 60% in the liver with respect to the commercial sample, which suggests that this product could be a suitable contrast agent for NMR imaging of liver and spleen. PMID- 15878376 TI - Steady-state diffusion of water through soft-contact-lens materials. AB - Water transport through soft contact lenses (SCL) is important for acceptable performance on the human eye. Chemical-potential gradient-driven diffusion rates of water through SCL materials are measured with an evaporation-cell technique. Water is evaporated from the bottom surface of a lens membrane by impinging air at controlled flow rate and humidity. The resulting weight loss of a water reservoir covering the top surface of the contact-lens material is recorded as a function of time. New results are reported for a conventional hydrogel material (SofLens One Day, hilafilcon A, water content at saturation w10 = 70 weight %) and a silicone hydrogel material (PureVision, balafilcon A, w10 = 36%), with and without surface oxygen plasma treatment. Also, previously reported data for a conventional 2-hydroxyethyl methacrylate (HEMA)-SCL (w10 = 38%) hydrogel are reexamined and compared with those for SofLens One Day and PureVision hydrogels. Measured steady-state water fluxes are largest for SofLens One Day, followed by PureVision and HEMA. In some cases, the measured steady-state water fluxes increase with rising relative air humidity. This increase, due to an apparent mass-transfer resistance at the surface (trapping skinning), is associated with formation of a glassy skin at the air/membrane interface when the relative humidity is below 55-75%. Steady-state water fluxes are interpreted through an extended Maxwell-Stefan diffusion model for a mixture of species starkly different in size. Thermodynamic nonideality is considered through Flory-Rehner polymer-solution theory. Shrinking/swelling is self-consistently modeled by conservation of the total polymer mass. Fitted Maxwell-Stefan diffusivities increase significantly with water concentration in the contact lens. PMID- 15878377 TI - The effect of the antimicrobial peptide, Dhvar-5, on gentamicin release from a polymethyl methacrylate bone cement. AB - The objective of this study was to investigate the release mechanism and kinetics of the antimicrobial peptide, Dhvar-5, both alone and in combination with gentamicin, from a standard commercial polymethyl methacrylate (PMMA) bone cement. Different amounts of Dhvar-5 were mixed with the bone cement powders of Osteopal and the gentamicin-containing Osteopal G bone cement and their release kinetics from the polymerized cement were investigated. Additionally, the internal structure of the bone cements were analysed by scanning electron microscopy (SEM) of the fracture surfaces. Secondly, porosity was investigated with the mercury intrusion method and related to the observed release profiles. In order to obtain an insight into the mechanical characteristics of the bone cement mixtures, the compressive strength of Osteopal and Osteopal G with Dhvar-5 was also investigated. The total Dhvar-5 release reached 96% in the 100 mg Dhvar 5/g Osteopal cement, whereas total gentamicin release from Osteopal G reached only 18%. Total gentamicin release increased significantly to 67% with the addition of 50mg Dhvar-5/g, but the Dhvar-5 release was not influenced. SEM showed an increase of dissolved gentamicin crystals with the addition of Dhvar-5. The mercury intrusion results suggested an increase of small pores (< 0.1 microm) with the addition of Dhvar-5. Compressive strength of Osteopal was reduced by the addition of Dhvar-5 and gentamicin, but still remained above the limit prescribed by the ISO standard for clinical bone cements. We therefore conclude that the antimicrobial peptide, Dhvar-5, was released in high amounts from PMMA bone cement. When used together with gentamicin sulphate, Dhvar-5 made the gentamicin crystals accessible for the release medium presumably through increased micro porosity (< 0.1 microm) resulting in a fourfold increase of gentamicin release. PMID- 15878378 TI - Surface modification of biodegradable polyesters with fatty acid conjugates for improved drug targeting. AB - We describe a general method for incorporating target ligands into the surface of biocompatible polyester poly(lactic-co-glycolic acid) (PLGA) 50/50 materials using fatty acids. Avidin-fatty acid conjugates were prepared and efficiently incorporated into PLGA. Avidin was chosen as an adaptor protein to facilitate the attachment of a variety of biotinylated ligands. We show that fatty acid preferentially associates with the hydrophobic PLGA matrix, rather than the external aqueous environment, facilitating a prolonged presentation of avidin over several weeks. We successfully applied this approach in both microspheres encapsulating a model protein, bovine serum albumin, and PLGA scaffolds fabricated by a salt-leaching method. Because of its ease, generality and flexibility, this strategy promises widespread utility in modifying the surface of PLGA-based materials for applications in drug delivery and tissue engineering. PMID- 15878379 TI - Chitosan beads as molecularly imprinted polymer matrix for selective separation of proteins. AB - Two kinds of molecularly imprinted polymers were prepared using hemoglobin as the imprinting molecule, acrylamide as the functional monomer, chitosan beads and maleic anhydride-modified chitosan beads as matrixes, respectively. Static adsorbing experimental results showed that an equal class of adsorption was formed in the imprinted polymers and the adsorption equilibrium constant and the maximum adsorption capacity were evaluated. Chromatographic characteristics showed that the column bedded with the hemoglobin imprinted beads could separate hemoglobin and bovine serum albumin effectively from their mixture, which indicates that the imprinted beads have very higher selectivity for hemoglobin than the non-imprinted with the same chemical composition. PMID- 15878380 TI - Collection of neural inducing factors from PA6 cells using heparin solution and their immobilization on plastic culture dishes for the induction of neurons from embryonic stem cells. AB - Embryonic stem (ES) cells have the ability to replicate themselves and differentiate into various mature cells. Recently, dopaminergic neurons were efficiently induced from ES cells using mouse stromal cells (PA6 cells) as a feeder cell layer. This simple procedure seems to be very efficient to obtain dopamine-releasing cells for future clinical cell transplantation treatment of Parkinson's disease. In this study, we prepared stock solutions containing neural inducing factors (NIFs) by washing PA6 cells with phosphate-buffered saline containing heparin. ES cells grew successfully in culture media supplemented with 33 v/v% NIFs stock solution, and the rate of neural differentiation of ES cell progeny increased with increasing heparin concentration in the culture media. In addition, NIFs-immobilized surfaces were prepared by exposing polyethyleneimine modified surfaces to NIFs stock solutions. The NIFs-immobilized culture dish effectively supported cell growth as the culture medium supplemented with NIFs stock did, but its induction effect to dopaminergic neurons from ES cells was much smaller than free NIFs. NIFs stock solutions have two different activities. One can stimulate cell growth and the other induces differentiation of ES cells to the neural fate when heparin existed. The former factors were effectively immobilized on the culture dish, but those that induce differentiation may not be. Further optimization is required. PMID- 15878381 TI - The role of luteinizing hormone-releasing hormone therapy in locally advanced prostate cancer and biochemical failure: considerations for optimal use. AB - BACKGROUND: More patients are being diagnosed with prostate cancer at an earlier age with earlier stage disease because of advances in screening and detection. Investigators continue to explore the use of hormone therapy, particularly luteinizing hormone-releasing hormone (LHRH) analogues, earlier in the course of disease. OBJECTIVE: This review summarizes clinical evidence regarding the safety and efficacy of LHRH analogues in the treatment of locoregional disease or following biochemical failure. METHODS: Relevant information from clinical studies was identified through a MEDLINE search of the medical literature published in English in the last 5 years (search terms: LHRH and prostate cancer). The search included prospective and retrospective clinical studies on LHRH therapy in locally advanced prostate cancer. Additional relevant publications published before 1999 were identified from citations in the resulting articles. RESULTS: The available clinical evidence suggests that the use of LHRH analogues as adjuvant or neoadjuvant therapy in conjunction with radiation therapy may improve survival outcomes. Few studies have evaluated the use of LHRH analogues after biochemical failure. However, several related studies indicate that initiating hormone therapy earlier rather than later may provide some clinical benefit. When considering early initiation of LHRH therapy, the potential risks of long-term treatment must be considered. Physiologic changes, such as deterioration of body composition and bone quality, may have important effects on the risk of developing cardiovascular disease and osteoporosis. CONCLUSIONS: Although some clinical evidence supports the use of LHRH analogues as adjuvant or neoadjuvant therapy or following biochemical failure, further study is needed. In the meantime, clinicians should carefully weigh the potential benefits and risks of early hormone therapy when making treatment decisions. PMID- 15878382 TI - Testim 1% testosterone gel for the treatment of male hypogonadism. AB - OBJECTIVE: The aim of this work was to review the pharmacokinetic and clinical profile of Testim (Auxilium Pharmaceuticals, Norristown, Pennsylvania) 1% gel formulation of testosterone for the treatment of male hypogonadism. METHODS: An English-language search of the medical literature was conducted using PubMed (1998-December 2004) and EMBASE (1998-December 2004). Search terms included ag(e)ing male, male hypogonadism, late-onset hypogonadism, testosterone, testosterone deficiency, testosterone therapy, testosterone replacement therapy, androgen therapy, testosterone gel, and Testim. Bibliographies of retrieved articles were also reviewed. RESULTS: Five published clinical studies were reviewed. Testim 50 mg showed clear pharmacokinetic differences from AndroGel (known as Testogel in Europe; Unimed Pharmaceuticals, Inc., and Solvay Pharmaceuticals, Inc., Marietta, Georgia) 1% testosterone gel 50 mg, with increases of 30% (90% CI, 8%-57%) and 47% (90% CI, 20%-79%) versus AndroGel, respectively, in AUC(0-24h) for total serum testosterone and free testosterone. In a 30-day study of 638 men with hypogonadism, sexual desire scores and sexual motivation scores increased after Testim treatment at weeks 1, 2, 3, and 4 (each, P < 0.001). During 12 months of treatment with Testim 50 or 100 mg in 371 men with hypogonadism, total serum testosterone levels were raised to and maintained within the normal adult range, lean body mass increased by 2.2 kg (P < 0.001), fat mass fell by 2.1% (P < 0.001), and bone mineral density increased by 2.58% (P < 0.001). Mean scores for sexual desire, performance, motivation, and spontaneous erections were all significantly higher (all, P < 0.001) than at baseline for all time points during 12-month studies of Testim. In 2 studies comparing Testim with different testosterone patches, treatment with the gel resulted in 10-fold fewer application-site reactions than either patch. CONCLUSIONS: In men with hypogonadism, Testim gel raised and maintained serum testosterone levels to within the normal adult range, alleviated signs and symptoms associated with hypogonadism, and was well tolerated. PMID- 15878383 TI - Potential use of biochemical markers of bone turnover for assessing the effect of calcium supplementation and predicting fracture risk. AB - BACKGROUND: Biochemical markers of bone remodeling have been extensively used (independent of bone mass measurements) to document the efficacy of various anticatabolic and anabolic bone-modifying medications in reducing fracture risk. Nonetheless, their usefulness in determining the effectiveness of osteoporosis prevention and treatment, particularly calcium supplementation, has not been well established. OBJECTIVE: This article reviews the use of biochemical markers of bone remodeling as a measure of the effect of calcium supplementation and the implications for prediction of fracture risk. METHODS: A generalized search of MEDLINE from 1966 through April 2004 using the terms osteoporosis, fracture risk, and the specific bone biomarkers of interest was conducted to identify articles relating to these biomarkers and their relationship to prediction of fracture risk. A second MEDLINE search for the same period used the terms calcium, biological markers, and fracture risk to identify studies of calcium supplementation and bone biomarkers. In both cases, the reference lists of identified review articles were searched for additional publications. RESULTS: : Several biochemical markers of bone remodeling have been shown to be positively correlated with bone mineral density and fracture risk. Furthermore, calcium supplementation has been shown to have a significant correlation with levels of a number of these biomarkers (P < 0.05): the markers of bone formation procollagen type I carboxy and amino terminal peptides and serum bone-specific alkaline phosphatase, and the markers of bone resorption urinary hydroxyproline, urinary pyridinoline, urinary deoxypyridinoline, urinary amino terminal crosslinked telopeptide, and urinary and serum carboxy terminal crosslinked telopeptide. CONCLUSIONS: Calcium supplementation has a significant effect on a number of biomarkers of bone remodeling, an effect that is, in turn, correlated with decreased fracture risk. Most studies of the efficacy of calcium supplementation in reducing bone remodeling and influencing bone mineral density and fracture risk have involved calcium carbonate, although a few studies have found that other calcium salts may produce similar results. PMID- 15878384 TI - Efficacy and tolerability of the switch from a branded to a generic warfarin sodium product: an observer-blinded, randomized, crossover study. AB - BACKGROUND: Anticoagulation is an essential part of the management of numerous cardiovascular conditions. Although warfarin sodium has been in clinical use for >40 years, only 1 proprietary warfarin preparation was in use for long-term anticoagulation in Taiwan until 2001, when the Taiwan Food and Drug Administration approved a new generic formulation. OBJECTIVE: This study evaluated the efficacy and tolerability of switching from an established dose of the branded warfarin sodium product (in mg/wk) to the same dose of the generic product. METHODS: This was a randomized, observer-blinded,crossover study in Taiwanese patients who received a mechanical heart valve to replace an original valve or a previously implanted mechanical valve from March 2003 to August 2004 and had received branded warfarin therapy for >/=2 months postoperatively, with a mean baseline international normalized ratio (INR) in the range from 1.7 to 3.0 in the 8- to 12-week run-in period. Patients were randomized to 1 of 2 treatment sequences, each involving three 28-day periods: the generic product for 1 period followed by the branded product for 2 periods, or the branded product for 1 period followed by the generic product for 2 periods. Prothrombin time and INR were measured, and anticoagulant activity was compared. At each visit, treatment emergent adverse events were recorded and evaluated by the blinded observer, an internal medicine physician who was not involved in patients' care. RESULTS: Thirty-five patients were enrolled (25 men, 10 women; mean age, 52.4 years [range, 34.5-74.4 years]). All but 2 of the patients had received a replacement for a natural heart valve. Thirty-four patients (16 who received the generic product initially, 18 who received the branded product initially) completed the study without a dose change and were >/=75% compliant with therapy. There was no difference in pooled mean (SE) INR between the 2 products (2.28 [0.06] and 2.27 [0.06], respectively). The 90% CI for the difference was 96.4 to 104.9, well within the range for bioequivalence (80-120). There were no differences in the adverse-event profiles of the 2 formulations. CONCLUSIONS: No therapeutic inequivalence was demonstrated in this small, single-blind study, suggesting that the generic and branded warfarin products studied were equally effective in maintaining anticoagulation in patients with mechanical prosthetic heart valves. The 2 products had similar safety profiles. PMID- 15878385 TI - A retrospective chart review of the clinical effects of atypical antipsychotic drugs on glycemic control in institutionalized patients with schizophrenia and comorbid diabetes mellitus. AB - BACKGROUND: The association between schizophrenia and onset of type 2 diabetes mellitus (DM) is well documented. It is unclear whether this association is due to inherent risk factors in individuals with schizophrenia or to treatment with antipsychotic drugs, particularly atypical antipsychotic agents. OBJECTIVES: The goals of this retrospective, observational study were the following: (1) to determine whether adequate glycemic control could be achieved in patients with schizophrenia and comorbid DM who were undergoing treatment with atypical antipsychotic drugs, (2) to identify inherent risk factors that may affect glycemic control in this patient population, and (3) to consider the possibility that combined medication and inherent risk factors may affect glycemic control. METHODS: This was a retrospective, observational chart review that evaluated institutionalized patients in a New Jersey mental institution with concurrent diagnoses of schizophrenia and type 1 or 2 DM who were being treated with atypical antipsychotic agents (ie, olanzapine, risperidone, clozapine, or quetiapine) and were referred by their hospital internists to a weekly, half-day, in-hospital DM clinic. All patients with initial and end-point data for the efficacy measure (ie, change in glycosylated hemoglobin [HbA(1c)]) were included in the analysis. Mixed-effects linear and least-squares models were used to test whether a specific comorbidity had an effect on the change in HbA(1c) adjusted for the duration of the observation. RESULTS: A total of 72 patients met entry criteria. Among the 38 patients with baseline and end-point data (20 treated with olanzapine and 18 treated with risperidone), mean (SD) HbA(1c) decreased from 8.21% (2.4%) to 7.62% (1.7%). The only baseline demographic characteristic or comorbidity that predicted significant worsening in the adjusted HbA(1c) change was hepatitis (P = 0.003). CONCLUSIONS: Using appropriate, aggressive antidiabetic therapy, glycemic control was achieved in this group of patients with schizophrenia and comorbid DM who were treated with atypical antipsychotic agents. PMID- 15878386 TI - Classification and occurrence of clinically significant drug interactions with irinotecan and oxaliplatin in patients with metastatic colorectal cancer. AB - BACKGROUND: Pharmacokinetic and pharmacodynamic drug interactions with cytotoxic drugs may significantly influence the efficacy and toxicity of chemotherapy. OBJECTIVE: The purpose of this study was to identify drug interactions with irinotecan and oxaliplatin reported in the literature, to assess their clinical significance, and to examine the occurrence of these interactions in patients with metastatic colorectal cancer treated with either irinotecan or oxaliplatin or both. METHODS: To obtain data on drug-drug interactions with irinotecan and oxaliplatin, a literature search of PubMed and EMBASE was conducted using the search terms irinotecan, oxaliplatin, and interactions (English-language studies only published between 1980 and August 2004). The interactions found were subsequently classified for documentation evidence and severity of clinical effect, according to a 5-level classification system of a standard reference text, by a study panel of medical oncologists and clinical pharmacists. Comedication of patients who were treated with irinotecan or oxaliplatin, or both, was then examined to determine the occurrence of clinically significant interactions. RESULTS: Ninety-eight patients (50 women, 48 men;mean age, 60 years) were included in the study. Seventeen interactions with irinotecan were found in the literature, and 11 were classified as clinically significant. Only 1 nonspecific, clinically significant interaction was identified for oxaliplatin. Irinotecan-treated patients received a mean of 8 different comedications and oxaliplatin-treated patients received a mean of 6. Apart from antiemetic and antidiarrheal drugs that were prescribed for treatment-related toxicities, only 1 patient appeared to be exposed to a possible clinically significant interaction (between irinotecan and phenytoin). CONCLUSIONS: Eleven of the 17 interactions with irinotecan that were found in the literature were classified as clinically significant versus 1 clinically significant interaction with oxaliplatin. The occurrence of these interactions in the study patients with metastatic colorectal cancer was low. For medication surveillance purposes, however, the significant interactions should be considered in clinical practice. PMID- 15878387 TI - A patient-tailored N-acetylcysteine protocol for acute acetaminophen intoxication. AB - BACKGROUND: Hepatotoxicity as a result of acetaminophen(APAP) intoxication has become an important problem, but early intervention with N-acetylcysteine (NAC) is effective in preventing hepatic injury. Two NAC regimens are currently approved for acute APAP intoxication: NAC administered orally every 4 hours for 72 hours, and NAC administered intravenously for 20 hours within 8 to 10 hours after ingestion of a potentially hepatotoxic amount of APAP. However, clinical observations suggest that a variable treatment duration may be more appropriate than use of these predetermined, fixed-duration protocols. OBJECTIVES: This study investigated the tolerability and efficacy of a patient-tailored NAC protocol for acute APAP intoxication by comparing the incidence of hepatotoxicity in patients receiving this protocol and in historical controls receiving 1 of 2 fixed duration protocols: oral NAC for 72 hours and intravenous NAC for 20 hours within 8 to 10 hours after ingestion of a potentially hepatotoxic amount of APAP. METHODS: This was a retrospective case series study that included all patients admitted through the emergency department (ED) of the National Taiwan University Hospital with a diagnosis of APAP intoxication between October 1997 and October 2002. According to the patient-tailored protocol, which had been used in the ED since 1997, patients with a serum APAP concentration above the limit for possible risk based on a modified Rumack-Matthew nomogram received oral treatment with NAC 140 mg/kg, followed by maintenance doses of 70 mg/kg every 4 hours. NAC treatment was discontinued when the APAP concentration was <10 mg/L and serum aspartate aminotransferase (AST) was <40 IU/L. For the purposes of assessing clinical outcomes, patients were divided into 3 groups based on duration of treatment: the short-course group (/=73 hours). The primary outcome measure was development of hepatotoxicity, defined as a serum AST or alanine aminotransferase concentration >1000 IU/L. RESULTS: Twenty-seven patients were included in the study, 17 in the short-course group, 4 in the intermediate-course group, and 6 in the long-course group. The mean (SD) durations of NAC treatment in the respective groups were 22.1 (5.5) hours, 45.0 (8.2) hours, and 97.3 (33.2) hours. All 6 patients (22%) in the long-course group had hepatotoxicity (peak AST range, 1083 9770 IU/L); their treatment duration ranged from 80 to 164 hours. No patients in the short- or intermediate-course group had evidence of hepatotoxicity. One woman in the long-course group in whom initiation of NAC treatment was delayed by 28 hours died of fulminant hepatic failure. The overall incidence of hepatotoxicity was similar to that in the historical controls. CONCLUSIONS: In this retrospective case series inpatients who received patient-tailored NAC therapy for acute APAP intoxication, the incidence of hepatotoxicity was low and comparable to that in historical controls who received treatment with 1 of 2 fixed-duration regimens. Use of this protocol may have the potential to shorten hospital stays without increasing the risk to patients. However, the sample size was small, and the findings require confirmation in prospective clinical trials. PMID- 15878388 TI - Must an interest be a conflict? PMID- 15878389 TI - A comparison of the risk of hospitalizations due to chronicobstructive pulmonary disease in medicaid patients with various medication regimens, including ipratropium, inhaled corticosteroids, salmeterol, or their combination. AB - OBJECTIVE: The aim of this study was to compare the risk of hospitalizations related to chronic obstructive pulmonary disease (COPD) among Medicaid patients prescribed various medication regimens. METHODS: This was an observational, retrospective study set in the Texas Medicaid program. Eligible patients were aged 40 to 65 years, had a primary or secondary diagnosis of COPD, and had >/=1 prescription for ipratropium (IPR), inhaled corticosteroids (ICS), or salmeterol (SAL) between January 1, 1998, and August 31, 2000. Five index therapy groups were included in the risk analysis: IPR alone, ICS alone, SAL alone, ICS + IPR, and ICS + SAL. RESULTS: A total of 4447 patients were included in the study (IPR alone, n = 2435; ICS alone, n = 1088; SAL alone, n = 299; ICS + IPR, n = 410; and ICS + SAL, n = 215). After adjusting for baseline characteristics, ICS + SAL was associated with a 35% lower risk of COPD-related hospitalization (hazard ratio [HR], 0.653 [95% CI, 0.428-0.997]) versus IPR alone. ICS alone was associated with a 16% lower risk (HR, 0.844 [95% CI, 0.693-1.028]) and SAL alone was associated with a 24% lower risk (HR, 0.756 [95% CI, 0.539-1.060]) versus IPR alone, but neither of these was statistically significant. There was no decrease in risk with ICS + IPR versus IPR alone (HR, 1.111 [95% CI, 0.870-1.420]). Variables that indicated increased risk were as follows: increasing age (HR, 1.015 [95% CI, 1.003-1.027]); number of preindex emergency department visits (HR, 1.189 [95% CI, 1.080-1.309]); number of preindex hospitalization visits (HR, 1.342 [95% CI, 1.220-1.477] ); number of nonrespiratory comorbid diagnoses (HR, 1.046 [95% CI, 1.012-1.081]); and having a diagnosis of influenza/pneumonia (HR, 1.276 [95% CI, 1.062-1.533]) or other respiratory diseases (HR, 1.356 [95% CI, 1.134-1.622]). Comorbid asthma was not associated with increased risk. CONCLUSIONS: ICS + SAL was associated with a significantly lower risk of COPD related hospitalization compared with IPR alone during the initial 12 months of therapy in a Medicaid population. Additional studies are needed to confirm these findings across different populations. PMID- 15878390 TI - Public-released version of the Adverse Event Reporting System (AERS) database. PMID- 15878392 TI - Uptake of LPS/E. coli/latex beads via distinct signalling pathways in medfly hemocytes: the role of MAP kinases activation and protein secretion. AB - In response to LPS/E. coli treatment, extracellular signal-regulated kinase (ERK) is activated in medfly hemocytes. To explore the molecular mechanisms underlying LPS/E. coli/latex beads endo- and phagocytosis, we studied the signalling pathways leading to p38 and c-jun N-terminal kinase (JNK) activation. JNK and p38 like proteins were initially identified within medfly hemocytes. Flow cytometry analysis revealed that mitogen-activated protein kinases (MAPK) are required for phagocytosis. Inhibition of specific MAPK signalling pathways, with manumycin A, toxin A, cytochalasin D and latrunculin A, revealed activation of p38 via Ras/Rho/actin remodelling pathway and activation of JNK that was independent of actin cytoskeleton reorganization. ERK and p38 pathways, but not JNK, appeared to be involved in LPS-dependent hemocyte secretion, whereas all MAPK subfamilies seemed to participate in E. coli-dependent secretion. In addition, flow cytometry experiments in hemocytes showed that the LPS/E. coli-induced release was a prerequisite for LPS/E. coli uptake, whereas latex bead phagocytosis did not depend on hemocyte secretion. This is a novel aspect, as in mammalian monocytes/macrophages LPS/E. coli-triggered release has not been yet correlated with phagocytosis. It is of interest that these data suggest distinct mechanisms for the phagocytosis of E. coli and latex beads in medfly hemocytes. PMID- 15878393 TI - Identification of a novel phosphorylation site in ataxin-1. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal dominant neurodegenerative disease resulting from an expanded CAG repeat in the SCA1 gene that leads to an expanded polyglutamine tract in the gene product. Previous studies have demonstrated that serine at site 776 is phosphorylated [E.S. Emiamian, M.D. Kaytor, L.A. Duvick, T. Zu, S.K. Tousey, H.Y. Zoghbi, H.B. Clark, H.T. Orr, Serine 776 of ataxin-1 is critical for polyglutamine-induced disease in SCA1 transgenic mice, Neuron 38 (2003) 375-387.]. Studies of ataxin-1 S776 and serine mutated to an alanine, A776, have also shown differential protein-protein interactions and reduced neurodegeneration [H.K. Chen, P. Fernandez-Funez, S.F. Acevedo, Y.C. Lam, M.D. Kaytor, M.H. Fernandez, A. Aitken, E.M. Skoulakis, H.T. Orr, J. Botas, H.Y. Zoghbi, Interaction of Akt_phosphorylated ataxin-1 with 14-3 3 mediates neurodegeneration in spinocerebellar ataxia type 1.]. However, mutation of the site serine 776 to an alanine did not abolish all phosphorylation of the protein ataxin-1, suggesting the presence of additional phosphorylation sites [E.S. Emiamian, M.D. Kaytor, L.A. Duvick, T. Zu, S.K. Tousey, H.Y. Zoghbi, H.B. Clark, H.T. Orr, Serine 776 of ataxin-1 is critical for polyglutamine induced disease in SCA1 transgenic mice, Neuron 38 (2003) 375-387.]. Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and mutational analysis demonstrated a novel phosphorylation site at serine 239 of ataxin-1. PMID- 15878394 TI - Mitochondria are intracellular magnesium stores: investigation by simultaneous fluorescent imagings in PC12 cells. AB - To determine the nature of intracellular Mg2+ stores and Mg2+ release mechanisms in differentiated PC12 cells, Mg2+ and Ca2+ mobilizations were measured simultaneously in living cells with KMG-104, a fluorescent Mg2+ indicator, and fura-2, respectively. Treatment with the mitochondrial uncoupler, carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone (FCCP), increased both the intracellular Mg2+ concentration ([Mg2+]i) and the [Ca2+]i in these cells. Possible candidates as intracellular Mg2+ stores under these conditions include intracellular divalent cation binding sites, endoplasmic reticulum (ER), Mg-ATP and mitochondria. Given that no change in [Mg2+]i was induced by caffeine application, intracellular IP3 or Ca2+ liberated by photolysis, it appears that no Mg2+ release mechanism thus exists that is mediated via the action of Ca2+ on membrane-bound receptors in the ER or via the offloading of Mg2+ from binding sites as a result of the increased [Ca2+]i. FCCP treatment for 2 min did not alter the intracellular ATP content, indicating that Mg2+ was not released from Mg-ATP, at least in the first 2 min following exposure to FCCP. FCCP-induced [Mg2+]i increase was observed at mitochondria localized area, and vice versa. These results suggest that the mitochondria serve as the intracellular Mg2+ store in PC12 cell. Simultaneous measurements of [Ca2+]i and mitochondrial membrane potential, and also of [Ca2+]i and [Mg2+]i, revealed that the initial rise in [Mg2+]i followed that of mitochondrial depolarization for several seconds. These findings show that the source of Mg2+ in the FCCP-induced [Mg2+]i increase in PC12 cells is mitochondria, and that mitochondrial depolarization triggers the Mg2+ release. PMID- 15878395 TI - Calcyclin (S100A6) expression is stimulated by agents evoking oxidative stress via the antioxidant response element. AB - Calcyclin (S100A6) is a cell-specific, calcium binding protein of the S100 family whose expression is augmented in many types of cancer. By means of luciferase activity assays, RT-PCR and Northern blot hybridization, we established that transcription of S100A6 gene is increased by agents known to evoke oxidative stress. Mutation of the antioxidant response element (ARE) located at position 290/-281 of the calcyclin gene promoter, and overlapping the E-box sequence recognized by the upstream stimulatory factor (USF), led to inhibition of calcyclin gene promoter activity stimulated by cadmium ions. Electrophoretic mobility shift assays (EMSA) with the -302/-260 calcyclin gene promoter fragment revealed, apart from USF binding, the presence of another protein complex (N) shown by competitive EMSA to be bound to ARE. DNA affinity chromatography followed by Western blot showed the binding of Nrf2 transcription factor to the immobilized calcyclin gene promoter fragment and concomitant appearance of complex N in EMSA of the eluted fractions. The results indicate that agents evoking oxidative stress activate calcyclin gene via the ARE sequence in its promoter. PMID- 15878396 TI - Uneven cellular expression of recombinant alpha2A-adrenoceptors in transfected CHO cells results in loss of response in adenylyl cyclase inhibition. AB - Two populations of Chinese hamster ovary (CHO) cells expressing similar numbers of recombinant human alpha2A-adrenergic receptors (alpha2A-AR) showed different capacity to inhibit adenylyl cyclase (AC) activity. Cells transfected with an integrating vector exhibited agonist-dependent inhibition of forskolin-stimulated AC, whereas cells transfected with a non-integrating episomal vector showed no inhibition. Fluorescent microscopy and flow cytometry revealed a very uneven receptor distribution in the episomally transfected cell population. Monoclonal cell populations were expanded from this parent population. Most clones lacked significant amounts of receptors, while a few expressed receptors at high density; these exhibited efficient agonist-dependent inhibition of forskolin stimulated AC activity. Thus, dense receptor expression in only a few cells is not sufficient to evoke a significant inhibitory response in a functional assay where AC is stimulated in all cells. Consequently, a false negative result was produced. Furthermore, the cell population transfected with an integrating vector showed loss of homogeneity with increasing passage number. PMID- 15878397 TI - Bovine type I collagen inhibits Raw264.7 cell proliferation through phosphoinositide 3-kinase- and mitogen-activated protein kinase-dependent down regulation of cyclins D1, A and B1. AB - Bovine type I collagen (BIC), which is widely used as a fibrous extracellular matrix component in cell culture models, inhibits the progression of melanoma cell cycle via p27 up-regulation. BIC also induces nitric oxide synthase in macrophages through JunB/AP-1 and NF-kappaB activation. Given the previous observations, this study investigates the effect of BIC on the cell cycle progression and regulatory function of Raw264.7 macrophage cells and the responsible signaling pathways. Cell cycle analysis revealed that BIC completely suppressed proliferation of Raw264.7 cells with inhibition of the percentage of cells in the S phase and the reciprocal decrease in the G0/G1 phase. DNA synthesis was also inhibited by BIC, as evidenced by a decrease in the cellular incorporation of [3H]thymidine. The G1/S arrest induced by BIC was reversed by chemical inhibition of phosphatidylinositol 3-kinase (PI3-kinase) or overexpression of the p85 subunit of PI3-kinase. Either PD98059 or stable transfection with mitogen-activated protein kinase kinase-1 [MKK1(-)] or c-Jun N terminal kinase 1 [JNK1(-)] also released the cell cycle arrest. Immunoblot analyses revealed that the levels of cyclins D1, A and B1 were partly or completely down-regulated by BIC, but cyclin E, p21 and p27 were minimally changed. Chemical inhibition and dominant negative mutant overexpression experiments revealed that either PI3-kinase inhibition or JNK1(-) transfection prevented the decreases in cyclin D1, A and B1 by BIC, indicating that the PI3 kinase and JNK1 pathways were associated with disruption of the cyclins. The pathway involving MKK1-extracellular signal-regulated kinase-1/2 (ERK1/2) was responsible for the suppression of cyclin A and B1, but not that of cyclin D1. The present study showed that BIC inhibited proliferation of Raw264.7 cells and that the pathways involving PI3-kinase and mitogen-activated protein kinases regulate the cell cycle arrest. PMID- 15878398 TI - Increase of intracellular glutathione by low-level NO mediated by transcription factor NF-kappaB in RAW 264.7 cells. AB - The mechanism underlying the elevation of intracellular glutathione (GSH) in RAW 264.7 cells exposed to low concentrations of sodium nitroprusside (SNP), a well known nitric oxide (NO) donor, was investigated. The peak of intracellular GSH was reached at 6 h after exposure of the cells to SNP (0.1-0.5 mM), and this was preceded by the induction of mRNA for gamma-glutamylcysteine synthetase (gamma GCS; the rate-limiting enzyme of de novo GSH synthesis), which peaked at 3 h. N alpha-Tosyl-L-phenylalanine chloromethyl ketone (TPCK) and caffeic acid phenethyl ester (CAPE), specific inhibitors of NF-kappaB, significantly suppressed the SNP induced elevation of GSH protein and gamma-GCS mRNA, while curcumin, an inhibitor of AP-1, was less effective. Electrophoretic mobility shift assay (EMSA) showed that SNP exposure markedly increased the DNA binding of NF-kappaB, but not that of AP-1. Deletion or mutagenesis of the NF-kappaB site in the gamma-GCS gene promoter abolished the SNP-induced up-regulation of GSH protein and gamma-GCS mRNA. These results suggest that the elevation of intracellular GSH in RAW 264.7 cells exposed to low concentrations of SNP occurs through the operation of the de novo GSH pathway, and is mediated by transcriptional up-regulation of the gamma GCS gene, predominantly at the NF-kappaB binding site in its promoter. PMID- 15878399 TI - Outer membrane protein 25-a mitochondrial anchor and inhibitor of stress activated protein kinase-3. AB - Stress-activated protein kinase-3 (SAPK3) is unique amongst the mitogen-activated protein kinase (MAPK) family with its C-terminal 5 amino acids directing interaction with the PDZ domain-containing substrates alpha1-Syntrophin and SAP90/PSD95. Here, we identify three additional PDZ domain-containing binding partners, Lin-7C, Scribble, and outer membrane protein 25 (OMP25). This latter protein is localised together with SAPK3 at the mitochondria but it is not a SAPK3 substrate. Instead, OMP25 inhibits SAPK3 activity towards PDZ domain containing substrates such as alpha1-Syntrophin and substrates without PDZ domains such as the mitochondrial protein Sab. This is a new mechanism for the regulation of SAPK3 and suggests that its intracellular activity should not be solely assessed by its phosphorylation status. PMID- 15878400 TI - The plasticity of p19 ARF null hepatic stellate cells and the dynamics of activation. AB - In the healthy adult liver, quiescent hepatic stellate cells (HSCs) present the major site for vitamin A storage in cytoplasmic lipid droplets. During liver injury due to viral infection or alcohol intoxication, HSCs get activated and produce high amounts of extracellular matrix components for tissue repair and fibrogenesis. Employing p19 ARF deficiency, we established a non-transformed murine HSC model to investigate their plasticity and the dynamics of HSC activation. Primary HSCs isolated from livers of adult p19 ARF null mice underwent spontaneous activation through long-term passaging without an obvious replicative limit. The immortalized cell line, referred to as M1-4HSC, showed stellate cell characteristics including the expression of desmin, glial fibrillary acidic protein, alpha-smooth muscle actin and pro-collagen I. Treatment of these non-tumorigenic M1-4HSC with pro-fibrogenic TGF-beta1 provoked a morphological transition to a myofibroblastoid cell type which was accompanied by enhanced cellular turnover and impaired migration. In addition, M1-4HSCs expressed constituents of cell adhesion complexes such as p120(ctn) and beta catenin at cell borders, which dislocalized in the cytoplasm during stimulation to myofibroblasts, pointing to the epitheloid characteristics of HSCs. By virtue of its non-transformed phenotype and unlimited availability of cells, the p19(ARF) deficient model of activated HSCs and corresponding myofibroblasts render this system a highly valuable tool for studying the cellular and molecular basis of hepatic fibrogenesis. PMID- 15878401 TI - Physiological changes in Campylobacter jejuni on entry into stationary phase. AB - Campylobacter jejuni NCTC 11168 does not exhibit the general increase in cellular stress resistance on entry into stationary phase that is seen in most other bacteria. This is consistent with the lack of global stationary phase regulatory elements in this organism, deduced from an analysis of its genome sequence. We now show that C. jejuni NCTC 11168 does undergo certain changes in stationary phase, of a pattern not previously described. As cells entered stationary phase there was a change in membrane fatty acid composition, principally a decrease in the proportion of unsaturated fatty acids and an increase in the content of cyclopropane and short-chain fatty acids. These changes in membrane composition were accompanied by an increase in the resilience of the cell membrane towards loss of integrity caused by pressure and an increase in cellular pressure resistance. By contrast, there were no major changes in resistance to acid or heat treatment. A similar pattern of changes in stress resistance on entry into stationary phase was seen in C. jejuni NCTC 11351, the type strain. These changes appear to represent a restricted physiological response to the conditions existing in stationary phase cultures, in an organism having limited capacity for genetic regulation and adaptation to environment. PMID- 15878402 TI - A rapid chemiluminescent slot blot immunoassay for the detection and quantification of Clostridium botulinum neurotoxin type E, in cultures. AB - A simple, rapid, cost-effective in vitro slot blot immunoassay was developed for the detection and quantification of botulinum neurotoxin type E (BoNT/E) in cultures. Culture supernatants of 36 strains of clostridia, including 12 strains of Clostridium botulinum type E, 12 strains of other C. botulinum neurotoxin serotypes, and 12 strains of other clostridial species were tested. Samples containing BoNT/E were detected using affinity-purified polyclonal rabbit antisera prepared against BoNT/E with subsequent detection of secondary antibodies using chemiluminescence. All strains of C. botulinum type E tested positive, while all non C. botulinum type E strains tested negative. The sensitivity of the slot blot immunoassay for detection of BoNT/E was approximately four mouse lethal doses (MLD). The intensity of chemiluminescence was directly correlated with the concentration of BoNT/E up to 128 MLD, allowing quantification of BoNT/E between 4 and 128 MLD. The slot blot immunoassay was compared to the mouse bioassay for detection of BoNT/E using cultures derived from fish samples inoculated with C. botulinum type E, and cultures derived from naturally contaminated environmental samples. A total of 120 primary enrichment cultures derived from fish samples, of which 103 were inoculated with C. botulinum type E, and 17 were uninoculated controls, were assayed. Of the 103 primary enrichment cultures derived from inoculated fish samples, all were positive by mouse bioassay, while 94 were also positive by slot blot immunoassay, resulting in a 7.5% false-negative rate. All 17 primary enrichment cultures derived from the uninoculated fish samples were negative by both mouse bioassay and slot blot immunoassay. A total of twenty-six primary enrichment cultures derived from environmental samples were tested by mouse bioassay and slot blot immunoassay. Of 13 primary enrichment cultures positive by mouse bioassay, 12 were also positive by slot blot immunoassay, resulting in a 3.8% false-negative rate. All 13 primary enrichment cultures that tested negative by mouse bioassay also tested negative by slot blot immunoassay. The slot blot immunoassay could be used routinely as a positive screen for BoNT/E in primary enrichment cultures, and could be used as a replacement for the mouse bioassay for pure cultures. PMID- 15878403 TI - Purification, partial amino acid sequence and mode of action of pediocin PD-1, a bacteriocin produced by Pediococcus damnosus NCFB 1832. AB - Pediocin PD-1 is a ribosomally synthesized antimicrobial peptide produced by Pediococcus damnosus NCFB1832. It inhibits the growth of several food spoilage bacteria, including malolactic bacteria isolated from wine. Pediocin PD-1 is 2866.87+/-0.4 Da in size, has an isoelectric point (pI) of ca. 9.0 and, on amino acid composition, has partial homology to the lantibiotic plantaricin C. The highest activity of pediocin PD-1 against cells of Oenococcus oeni was observed at an external pH of 5.0 and at 25 degrees C. The primary mode of action of pediocin PD-1 is most probably due to pore formation, as indicated by the efflux of K+ from metabolically active cells of O. oeni. In the presence of 10 mM gadolinium (Gd3+), pediocin PD-1 did not affect cells of O. oeni. This suggests that the mode of action of pediocin PD-1 relies on a net negatively charged cell surface. In comparison to nisin, pediocin PD-1 is less active against non-growing cells of O. oeni. PMID- 15878404 TI - In vitro screening of probiotic properties of Saccharomyces cerevisiae var. boulardii and food-borne Saccharomyces cerevisiae strains. AB - The probiotic potential of 18 Saccharomyces cerevisiae strains used for production of foods or beverages or isolated from such, and eight strains of Saccharomyces cerevisiae var. boulardii, was investigated. All strains included were able to withstand pH 2.5 and 0.3% Oxgall. Adhesion to the nontumorigenic porcine jejunal epithelial cell line (IPEC-J2) was investigated by incorporation of 3H-methionine into the yeast cells and use of liquid scintillation counting. Only few of the food-borne S. cerevisiae strains exhibited noteworthy adhesiveness with the strongest levels of adhesion (13.6-16.8%) recorded for two isolates from blue veined cheeses. Merely 25% of the S. cerevisiae var. boulardii strains displayed good adhesive properties (16.2-28.0%). The expression of the proinflammatory cytokine IL-1alpha decreased strikingly in IPEC-J2 cells exposed to a Shiga-like toxin 2e producing Escherichia coli strain when the cells were pre- and coincubated with S. cerevisiae var. boulardii even though this yeast strain was low adhesive (5.4%), suggesting that adhesion is not a mandatory prerequisite for such a probiotic effect. A strain of S. cerevisiae isolated from West African sorghum beer exerted similar effects hence indicating that food borne strains of S. cerevisiae may possess probiotic properties in spite of low adhesiveness. PMID- 15878405 TI - Tracing Listeria monocytogenes isolates from cold-smoked salmon and its processing environment in Iceland using pulsed-field gel electrophoresis. AB - Listeria spp. and Listeria monocytogenes contamination of cold-smoked salmon (n=125) and its processing environment (n=522) were evaluated during surveys conducted in 1997-1998 and 2001 as well as in samples of final products analysed in 2001. The overall frequencies of Listeria spp. and L. monocytogenes in samples from all sources were 15.1% and 11.3%, respectively, but the incidence of L. monocytogenes in cold-smoked salmon final products was only 4%. A total of 201 L. monocytogenes isolates were characterised by Pulsed-Field Gel Electrophoresis (PFGE) in order to trace L. monocytogenes contamination in the processing plants. The combination of AscI and ApaI macrorestriction patterns yielded 24 different pulsotypes in 6 plants. One pulsotype observed by AscI restriction digestion comprised 148 of the 167 typed isolates from two processing plants. Two other pulsotypes predominated in samples from raw material, processing environments and final products. The results indicate that raw material, floors, and drains are potential sources of the L. monocytogenes found on cold-smoked salmon products. This highlights the need to readdress the design and cleaning of processing plants and equipment, and staff behavior. Hindering the introduction into and spread of the organism through the processing environment is necessary to avoid jeopardizing safety of the final product. PMID- 15878406 TI - Effect of prior growth temperature, type of enrichment medium, and temperature and time of storage on recovery of Listeria monocytogenes following high pressure processing of milk. AB - A five-isolate cocktail of Listeria monocytogenes (10(3) cfu/ml in skim or whole raw milk) was subjected to 450 MPa for 900 s or 600 MPa for 90 s. The effects of prior growth temperature, type of milk (skim vs. whole), type of recovery enrichment media (optimized Penn State University [oPSU] broth, Listeria Enrichment Broth [LEB], Buffered LEB [BLEB], Modified BLEB [MBLEB], and milk), storage temperature and storage time on the recovery of L. monocytogenes were examined. Optimized PSU broth significantly increased the recovery of L. monocytogenes following high pressure processing (HPP), and was 63 times more likely to recover L. monocytogenes following HPP, compared to LEB, BLEB and MBLEB broths (p<0.05; Odds Ratio=63.09, C.I. 23.70-167.96). There was a significant main effect for prior growth temperature (p<0.05). However, this relationship could not be interpreted given the significant interaction effects between temperature and both pressure and milk type. HPP-injured L. monocytogenes could be recovered using both LEB and oPSU broths after storage of milk at 4, 15 and 30 degrees C, with recovery being maximal after 24 to 72 h of storage; however, recovery yield dropped to 0% after prolonged storage of milk at 4 and 30 degrees C. In contrast, storage of milk at 15 degrees C yielded the most rapid rate of recovery and the highest recovery yield (100%), which remained high throughout the 14 days of storage at 15 degrees C. The above factors need to be taken into consideration when designing challenge studies to insure complete inactivation of L. monocytogenes and possibly other foodborne pathogens during high pressure processing of foods. PMID- 15878407 TI - Pulsed-field gel electrophoresis types of Campylobacter spp. in Danish turkeys before and after slaughter. AB - In this study, seven Danish turkey flocks were investigated at the farm, on arrival to the slaughterhouse, and during and after slaughter. Flocks were selected based on their Campylobacter spp. status at the farm and three Campylobacter negative and four Campylobacter positive flocks were included in the study. At the slaughterhouse, 70-75 samples were collected at different points from the shackling station to packaging of the final meat cuttings. Samples included cloacal swabs, neckskin, liver, heart, meat and environmental samples. Detection of Campylobacter was carried out by conventional culture and by the EiaFoss system (Foss Electric, Hilleroed, Denmark) for detecting Campylobacter spp. in food, using Preston Broth as enrichment medium. The two methods were compared and sensitivities and specificities were calculated using the conventional culture as gold standard. The three negative flocks were consistently negative from the farm and all through processing. Among the samples from the positive flocks, the frequency of positive samples obtained at the slaughterhouse varied. The frequency of positive samples obtained from the four positive flocks varied and was found to be 4%, 49%, 87% and 96%, respectively. In 31 out of 424 samples, discrepancies were observed between results obtained by the EiaFoss system and the conventional culture technique. The sensitivity for the EiaFoss system was calculated to be 0.94 for meat and neckskin samples. A total of 161 strains were genotyped by pulsed-field gel electrophoresis (PFGE) in order to investigate possible changes in carriage of Campylobacter spp. strains during processing. In three flocks, only one PFGE type was encountered in samples collected at the farm level and, in one flock, two different types were observed. In two flocks, the strain from the farm was also isolated in samples collected at the slaughterhouse. Changes in carriage were observed in two flocks during processing, in particular post chilling. PMID- 15878408 TI - Survival curves of heated bacterial spores: effect of environmental factors on Weibull parameters. AB - The classical D-value of first order inactivation kinetic is not suitable for quantifying bacterial heat resistance for non-log linear survival curves. One simple model derived from the Weibull cumulative function describes non-log linear kinetics of micro-organisms. The influences of environmental factors on Weibull model parameters, shape parameter "p" and scale parameter "delta", were studied. This paper points out structural correlation between these two parameters. The environmental heating and recovery conditions do not present clear and regular influence on the shape the parameter "p" and could not be described by any model tried. Conversely, the scale parameter "delta" depends on heating temperature and heating and recovery medium pH. The models established to quantify these influences on the classical "D" values could be applied to this parameter "delta". The slight influence of the shape parameter p variation on the goodness of fit of these models can be neglected and the simplified Weibull model with a constant p-value for given microbial population can be applied for canning process calculations. PMID- 15878409 TI - Practical application of dynamic temperature profiles to estimate the parameters of the square root model. AB - Optimal experimental design for parameter estimation (OED/PE) is a promising method to improve parameter estimation accuracy and minimise experimental effort in the field of predictive microbiology. In this paper, the OED/PE methodology was applied on two practical examples: the growth of Bacillus cereus and Enterobacter cloacae in liquid whole egg product. Both strains were recovered from samples of a commercial product. The goal of the modelling exercise was to quantify the influence of temperature on bacterial growth. The Baranyi-model for bacterial growth combined with the Ratkowsky square root model to describe temperature dependence was used. Using this model, a temperature step profile was calculated based on the optimal D-criterion. The model was then fitted against the experimental bacterial growth curve measured under the dynamic temperature conditions. This process was repeated until the parameters could be estimated with sufficient accuracy, apparent by the model prediction errors. For B. cereus, prior information could be extracted from the literature, allowing calculating a dynamic temperature profile directly. Two-step profiles were sufficient to obtain a good estimation for the model parameters. No prior information could be found for E. cloacae. Therefore, a limited series of static experiments had to be conducted to obtain usable prior model parameters estimates. Only one dynamic experiment was then needed to achieve a good estimation. PMID- 15878410 TI - Real-time PCR-based methods for detection of Mycobacterium avium subsp. paratuberculosis in water and milk. AB - A real-time PCR assay for quantitative detection of Mycobacterium avium paratuberculosis has been developed. It targets and amplifies sequences from the IS900 insertion element which is specific for this bacterium, and includes an internal amplification control. The assay was tested against 18 isolates of M. avium paratuberculosis, 17 other mycobacterial strains, and 25 non-mycobacterial strains, and was fully selective. It is capable of detecting <3 genomic DNA copies with 99% probability or alternatively, using cells directly in the reaction, 12 cells can be detected with 99% probability. Using prior centrifugation, the assay was able to consistently and quantifiably detect 10(2) M. avium paratuberculosis cells in 20 ml artificially contaminated drinking water. With a simple detergent and enzymatic sample pretreatment before centrifugation and nucleic acid extraction, the assay was able to consistently detect 10(2) M. avium paratuberculosis in 20 ml artificially contaminated semi skimmed milk. The assay will be a useful addition to the range of diagnostic tools available for the study of M. avium paratuberculosis. PMID- 15878411 TI - The effect of short-time microwave exposures on Escherichia coli O157:H7 inoculated onto chicken meat portions and whole chickens. AB - Small portions of fresh chicken breasts weighting 20 g each and fresh whole chickens, weighting on average 1310 g each, were inoculated with Escherichia coli O157:H7 (10(5)-10(6) cfu/g) and cooked, using two different domestic microwave ovens at full power. The chicken portions were heated for 5, 10, 15, 20, 25, 30, and 35 s and the whole chickens for 22 min. Following exposures, viable counts and temperature measurements were performed. Although the chicken breast portions looked well-cooked after 30 s of MW heating at a mean end-point surface temperature of 69.8 degrees C, a mean concentration of 83 cfu/g E. coli O157:H7 cells was recovered. Elimination of E. coli O157:H7 cells occurred only after 35 s of MW exposure at 73.7 degrees C. When whole chickens were thoroughly cooked by MW heating, the final subsurface temperatures, measured in the thighs and wings, ranged from 60.2 degrees C to 92 degrees C and viable cells of E. coli O157:H7 were recovered from all samples of whole chicken. The results indicate that short time exposures of chicken portions to microwave heating do not eliminate E. coli O157:H7. PMID- 15878412 TI - Antimicrobial effect of lactic acid producing bacteria culture condensate mixture (LCCM) against Salmonella enteritidis. AB - The antimicrobial effects of a lactic acid producing bacteria culture condensate mixture (LCCM) were assessed against Salmonella enteritidis. In the presence of LCCM, bacterial growth was assessed in vitro by the measurement of optical density (OD) and viable bacterial counting. At concentrations of 1.25 and 2.5% LCCM, OD values were significantly lower than that of the control broth, and at concentrations of 5 and 10% LCCM, OD values did not increase for the entire period of experiment. At 8 h after incubation, the viable bacterial numbers in 5% and 10% LCCM-containing broths were remarkably lower than that in the control broth. This antimicrobial ability of the LCCM was fundamentally attributed to causing cell death rather than inhibiting growth. Even when the pH of LCCM containing broth was adjusted to 7.2, the number of viable bacteria was significantly lower in the broths containing LCCM over 2.5% than that in control broth at 8 h after incubation. However, the OD value of each culture in the presence of each concentration of the LCCM increased over 1.0 at the completion of the experiment. The in vivo antimicrobial effects of the LCCM against S. enteritidis were also assessed. In S. enteritidis-infected mice, the LCCM decreased both the viable bacteria found in the feces and the mortality rate of the mice. These findings showed that the LCCM might have an antimicrobial ability against S. enteritidis. PMID- 15878413 TI - Occupational vehicular accident claims: a workers' compensation analysis of Oregon truck drivers 1990-1997. AB - This study used workers' compensation data from Oregon from 1990 to 1997 to examine workers' compensation claims from vehicular accidents by truck drivers, and to calculate claim rate estimates using baseline data derived from the US Bureau of Census' Current Population Surveys. During this period, 1168 valid injury claims due to vehicular accidents were filed representing an accident claim rate of 50.3 (95% CI: 45.1-55.5) per 10,000 truck drivers annually. There were 19 work-related vehicular accident fatalities recorded in the data over the 8-year period. Of all claimants, males constituted the majority (80.7%), most were 35 years of age or younger (51.4%) and had less than 1 year of job tenure (51.0%). Truck driver claim rates due to vehicular accidents were lowest during the 6 a.m. to 12 p.m. period. The average amount of compensable lost work days per injury claim was 57.8 days (S.D.=124.7) and the median claim time was 16.0 days with the inter-quartile range being 53.5 days. The amount of lost work reported increased with the claimant's age. A total of US$ 11,642,635 was paid in claims for vehicular accidents of truck drivers in Oregon over the time examined, which averaged US$ 9966 per claim, with a median claim amount of US$ 2590 and inter-quartile range of US$ 7670. Claims citing sprains were the most frequently recorded injury experienced from vehicular accidents. PMID- 15878414 TI - Radium isotopes in the Ulsan Bay. AB - In order to estimate the fluxes of Ra isotopes, we measured 224Ra, 226Ra, and 228Ra activities in the Ulsan Bay mixing zone. The convex upwards curvature of the plot of Ra isotope activities versus salinity for the mixing zone suggests that Ra isotopes are supplied from particles entering the mixing zone from both the river and bottom sediments. This addition increases the estuarine flux of 226Ra and 228Ra to the outer sea by factors of 15 and 95 over the flux attributable to Ra dissolved in the riverine water alone. In order to estimate the residence time of the water in Ulsan Bay, we applied a mass balance model to the distribution of 224Ra and 226Ra activities in the Ulsan Bay mixing zone with the inflow from the Taehwa River. The obtained residence times of the waters in the Ulsan Bay were estimated to be 6.8-11.4 d. The waters in the upper part of the estuary have long residence times whereas those in the lower part, in contact with the open sea, have shorter residence times. The mean residence time of the water in the Ulsan Bay was estimated to be 9.1 d. PMID- 15878415 TI - Decision making framework for application of forest countermeasures in the long term after the Chernobyl accident. AB - After the ChNPP accident a very large part of the territories covered by natural and artificial forests are contaminated with long-lived radionuclides, especially 137Cs. To protect people against exposure associated with forest contamination in the most affected regions of the NIS countries, countermeasures have been developed and recommended for the forest management. The paper presents a decision making framework to optimise forest countermeasures in the long term after the ChNPP accident. The approach presented is based on the analysis of the main exposure pathways and application of radiological, socio-economical and ecological criteria for the selection of optimal countermeasures strategies. Because of the diversity of these criteria modern decision support technologies based on multi-attributive analysis were applied. The results of the application of this approach are presented in a selected study area (Novozybkov district, Bryansk region, Russian Federation). The results prove and emphasize the need for a flexible technique to provide the optimised forest countermeasures taking into account radioecological, social and economic features of contaminated forests. PMID- 15878416 TI - Health and environmental impacts of a fertilizer plant--Part I: assessment of radioactive pollution. AB - The aim of the first part of this investigation is to assess the radioactive pollution caused by a production plant of complex fertilizers (that is to say containing nitrogen, phosphorus and, in some cases, potassium). Firstly, the authors determine the concentrations of natural radioactivity present in raw materials, end products and wastes of the industrial plant. Then, they carry out an assessment of radioactive releases into the atmosphere, hydrosphere and lithosphere as well as of their significance from the environmental point of view. The second part of the investigation will be aimed at assessing the annual effective doses to plant workers and to members of the population surrounding the industrial site. PMID- 15878417 TI - Health and environmental impacts of a fertilizer plant--Part II: assessment of radiation exposure. AB - In the previous paper the authors have studied the radioactive pollution caused by a complex fertilizers production plant. In this paper, the effective doses to the plant workers and to members of the population surrounding the industrial site are estimated. The authors have considered external irradiation, inhalation and ingestion of dust and inhalation of radon and radon daughters as the main occupational exposure routes. After estimating the single contributions, the total effective dose has been calculated as the sum of said contributions. Calculations have been differentiated according to the different tasks of the company employees. The estimated annual effective doses range from 0.6 to 1.4 mSv y(-1). Annual individual effective doses to local residents, resulting from internal and external irradiation caused by particulate matter emitted into the atmosphere by the plant have been estimated. The maximum individual dose rate is estimated to be about 4 microSv y(-1). PMID- 15878418 TI - Accumulation of Chernobyl-derived 137Cs in bottom sediments of some Finnish lakes. AB - The amount and vertical distribution of Chernobyl-derived 137Cs in the bottom sediments of some Finnish lakes were studied. Sediment and surface water samples were taken in 2000 and 2003 from 12 stations in nine lakes and the results were compared with those obtained in corresponding surveys carried out in 1969, 1978, 1988 and 1990. Each of the five deposition categories of Chernobyl fallout in Finland were represented. The depth profiles of 137Cs in the sediments showed considerable variety in the lakes studied. The peak values varied between 1.5 and 46 kBq kg(-1) dry wt. The size and shape of the peak did not always correlate with the amount of deposition in the area, but on the other hand, reflected differences in sedimentation processes in different lakes. In some of the lakes the peak still occurred in the uppermost (0-2 cm) sediment layer, but in an extreme case the peak occurred at a depth of 22-23 cm corresponding to a sedimentation rate of 16 mm year(-1) during the 14 years after the Chernobyl accident. The total amounts of 137Cs in sediments varied from 15 to 170 kBq m(-2) at the sampling stations studied. Since 1990, the amounts have continued to increase slightly in two lakes, but started to decrease in the other lakes. In most of the lakes, the total amounts of 137Cs in sediments were about 1.5-2 times higher than in local deposition. In two lakes, the ratio was below 1, but in one case 3.2. Compared with the total amounts of 137Cs at the same stations in the late 1960s and 1970s, the values were now at their highest, at about 60-fold. The most important factors affecting 137Cs values in sediments were the local amount of deposition and the type of the lake and the sediment, but in addition, there were a number of other factors to be considered. PMID- 15878419 TI - Soil to plant transfer of 238U, 226Ra and 232Th on a uranium mining-impacted soil from southeastern China. AB - Both soil and plant samples of nine different plant species grown in soils from southeastern China contaminated with uranium mine tailings were analyzed for the plant uptake and translocation of 238U, 226Ra and 232Th. Substantial differences were observed in the soil-plant transfer factor (TF) among these radionuclides and plant species. Lupine (Lupinus albus) exhibited the highest uptake of 238U (TF value of 3.7x10(-2)), while Chinese mustard (Brassica chinensis) had the least (0.5x10(-2)). However, in the case of 226Ra and 232Th, the highest TFs were observed for white clover (Trifolium pratense) (3.4x10(-2)) and ryegrass (Lolium perenne) (2.1x10(-3)), respectively. 232Th in the tailings/soil mixture was less available for plant uptake than 226Ra or 238U, and this was especially evident for Chinese mustard and corn (Zea mays). The root/shoot (R/S) ratios obtained for different plants and radionuclides shown that Indian mustard had the smallest R/S ratios for both 226Ra (5.3+/-1.2) and 232Th (5.3+/-1.7), while the smallest R/S ratio for 238U was observed in clover (2.8+/-0.9). PMID- 15878420 TI - Assessment of the radiological impacts of a zircon sand processing plant. AB - The paper presents the results of a study on radiological impacts resulting from a zircon sand processing plant located in the North-Eastern part of Italy. Activity concentrations of radionuclides found in materials associated with this industrial process are presented as well as the results of the assessment of the annual effective doses to the workers and the members of the public. Gamma spectrometric analyses were performed on "raw" sands, end-products, and soils sampled near the plant. Thermoluminescent dosimeters, electric pumps and electret ion chambers were used to measure the external irradiation, the indoor dust concentrations and the radon concentrations, respectively. The ground-level air concentration of radioactive particulate near the plant and the deposition of particulate matter were estimated by a Gaussian model (ISCLT3). Finally, the annual effective doses, calculated as provided for by Directive 96/29/Euratom, were estimated to be 1.7 mSv y(-1) for workers and 4.4 microSv y(-1) for members of the public. PMID- 15878421 TI - Seasonal variation of 210Pb activity concentration in outdoor air of Milan (Italy). AB - Weekly measurements of 210Pb activity concentration on aerosol particles were performed at ground level in outdoor air in Milan (Italy) during the years 2000 2001. The experimental method was based on the delayed alpha spectrometry of in grown 210Po activity detected on airborne particles collected on a filter support. On the same filter the 214Pb activity concentration was also measured during the sampling. 210Pb weekly concentration values showed a great variability and the average monthly values exhibited a seasonal pattern characterised by maxima in winter and minima in spring-summer. The same seasonal trend was also observed in 214Pb concentration values. 210Pb average yearly values were 0.70 and 0.82 mBq/m3 in 2000 and 2001, respectively, in good agreement with literature data registered at continental sites of similar latitude and longitude. The activity ratio 210Pb/214Pb was used to evaluate the residence time of fine aerosols in the urban air of Milan, which was 1-2 days. PMID- 15878422 TI - Energetics of primary and secondary electron transfer in Photosystem II membrane particles of spinach revisited on basis of recombination-fluorescence measurements. AB - Photon absorption by one of the roughly 200 chlorophylls of the plant Photosystem II (PSII) results in formation of an equilibrated excited state (Chl200*) and is followed by chlorophyll oxidation (formation of P680+) coupled to reduction of a specific pheophytin (Phe), then electron transfer from Phe- to a firmly bound quinone (QA), and subsequently reduction of P680+ by a redox-active tyrosine residue denoted as Z. The involved free-energy differences (DeltaG) and redox potentials are of prime interest. Oxygen-evolving PSII membrane particles of spinach were studied at 5 degrees C. By analyzing the delayed and prompt Chl fluorescence, we determined the equilibrium constant and thus free-energy difference between Chl200* and the [Z+,QA-] radical pair to be -0.43+/-0.025 eV, at 10 mus after the photon absorption event for PSII in its S(3)-state. On basis of this value and previously published results, the free-energy difference between P680* and [P680+,QA-] is calculated to be -0.50+/-0.04 eV; the free energy loss associated with electron transfer from Phe to QA is found to be 0.34+/-0.04 eV. The given uncertainty ranges do not represent a standard deviation or likely error, but an estimate of the maximal error. Assuming a QA /QA redox potential of -0.08 V, the following redox-potential estimates are obtained: +1.25 V for P680/P680+; +1.21 V for Z/Z+ (at 10 mus); -0.42 V for Phe /Phe; -0.58 V for P680*/P680+. PMID- 15878423 TI - Premicellar complexes of sphingomyelinase mediate enzyme exchange for the stationary phase turnover. AB - During the steady state reaction progress in the scooting mode with highly processive turnover, Bacillus cereus sphingomyelinase (SMase) remains tightly bound to sphingomyelin (SM) vesicles (Yu et al., Biochim. Biophys. Acta 1583, 121 131, 2002). In this paper, we analyze the kinetics of SMase-catalyzed hydrolysis of SM dispersed in diheptanoylphosphatidyl-choline (DC7PC) micelles. Results show that the resulting decrease in the turnover processivity induces the stationary phase in the reaction progress. The exchange of the bound enzyme (E*) between the vesicle during such reaction progress is mediated via the premicellar complexes (E(i)#) of SMase with DC7PC. Biophysical studies indicate that in E(i)# monodisperse DC7PC is bound to the interface binding surface (i-face) of SMase that is also involved in its binding to micelles or vesicles. In the presence of magnesium, required for the catalytic turnover, three different complexes of SMase with monodisperse DC7PC (E(i)# with i=1, 2, 3) are sequentially formed with Hill coefficients of 3, 4 and 8, respectively. As a result, during the stationary phase reaction progress, the initial rate is linear for an extended period and all the substrate in the reaction mixture is hydrolyzed at the end of the reaction progress. At low mole fraction (X) of total added SM, exchange is rapid and the processive turnover is limited by the steps of the interfacial turnover cycle without becoming microscopically limited by local substrate depletion or enzyme exchange. At high X, less DC7PC will be monodisperse, E(i)# does not form and the turnover becomes limited by slow enzyme exchange. Transferred NOESY enhancement results show that monomeric DC7PC in solution is in a rapid exchange with that bound to E(i)# at a rate comparable to that in micelles. Significance of the exchange and equilibrium properties of the E(i)# complexes for the interpretation of the stationary phase reaction progress is discussed. PMID- 15878424 TI - Effects of lipid composition on the membrane activity and lipid phase behaviour of Vibrio sp. DSM14379 cells grown at various NaCl concentrations. AB - The membrane lipid composition of living cells generally adjusts to the prevailing environmental and physiological conditions. In this study, membrane activity and lipid composition of the Gram-negative bacterium Vibrio sp. DSM14379, grown aerobically in a peptone-yeast extract medium supplemented with 0.5, 1.76, 3, 5 or 10% (w/v) NaCl, was determined. The ability of the membrane to reduce a spin label was studied by EPR spectroscopy under different salt concentrations in cell suspensions labeled with TEMPON. For lipid composition studies, cells were harvested in a late exponential phase and lipids were extracted with chloroform-methanol-water, 1:2:0.8 (v/v). The lipid polar head group and acyl chain compositions were determined by thin-layer and gas-liquid chromatographies. (31)P-NMR spectroscopy was used to study the phase behaviour of the cell lipid extracts with 20 wt.% water contents in a temperature range from 10 to 50 degrees C. The results indicate that the ability of the membrane to reduce the spin label was highest at optimal salt concentrations. The composition of both polar head groups and acyl chains changed markedly with increasing salinity. The fractions of 16:0, 16:1 and 18:0 acyl chains increased while the fraction of 18:1 acyl chains decreased with increasing salinity. The phosphatidylethanolamine fraction correlated inversely with the lysophosphatidylethanolamine fraction, with phosphatidylethanolamine exhibiting a minimum, and lysophosphatidylethanolamine a maximum, at the optimum growth rate. The fraction of lysophosphatidylethanolamine was surprisingly high in the lipid extracts. This lipid can form normal micellar and hexagonal phases and it was found that all lipid extracts form a mixture of lamellar and normal isotropic liquid crystalline phases. This is an anomalous behaviour since the nonlamellar phases formed by total lipid extracts are generally of the reversed type. PMID- 15878425 TI - Eight million babies die in the world each year. PMID- 15878426 TI - Women's views and experiences of postnatal hospital care in the Victorian Survey of Recent Mothers 2000. AB - OBJECTIVE: To investigate the views and experiences of postnatal hospital care of a representative sample of Victorian women who gave birth in Victoria, Australia, in 1999. DESIGN: Postal survey sent to women 5-6 months after giving birth. SETTING: Victoria, Australia. PARTICIPANTS: 1616 women who gave birth in Victoria in a 2-week period in September 1999. FINDINGS: 50.8% of women described their postnatal care in hospital as 'very good'. After adjusting for parity, method of birth, length of stay, model of care and socio-demographic characteristics, specific aspects of care with the greatest negative impact on the overall rating of postnatal care were as follows: midwives perceived as rushed and too busy (adjusted OR = 4.59 [95% CI 3.4-6.1]), doctors and midwives perceived as not 'always' sensitive and understanding (adjusted OR = 3.88 [2.8-5.5]), support and advice about going home not 'very helpful' (adjusted OR = 3.18 [2.3-4.5]), help and advice about baby feeding not 'extremely helpful' (adjusted OR = 3.27 [2.1 5.1]), not being given advice about baby feeding (adjusted OR = 2.84 [1.2-6.9]). Staying in hospital only 1-2 days (adjusted OR=2.00 [1.2-3.4]), and not knowing any of the midwives in the postnatal ward (adjusted OR = 1.80 [1.3-2.4]) were also associated with less positive ratings of postnatal hospital care. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: The 2000 Survey shows that women rate early postnatal care in hospital far less favourably than care in pregnancy, labour and birth. The findings indicate that interactions with caregivers are a major influence on women's overall rating of postnatal hospital care. Acting on these findings requires a greater focus on communication and listening skills, attention to staffing levels, and leadership promoting more women-centred care in postnatal wards. PMID- 15878427 TI - Chi2 test for linear trend--what's that? PMID- 15878428 TI - Mothers' lived experiences of co-care and part-care after birth, and their strong desire to be close to their baby. AB - OBJECTIVE: To describe how mothers of premature or sick mature babies, experienced the care and their own state of health after birth in postnatal care in a neonatal co-care ward. DESIGN: A Husserlian phenomenology method inspired by Giorgi was used. Six mothers were interviewed using a semi-structured, open-ended interview guide. SETTING: A neonatal ward using a concept of co-care for premature or sick mature babies and their mothers. FINDINGS: In essence, mothers felt that, whatever the circumstances, they wanted to be close to their babies. It was the mother's experience that the organisation, staff or other circumstances prolonged the separation from her baby. The mother experienced the separation from the baby intensely during the first days after birth (even for a short period of time); after returning home, they had still not come to terms with it. The mothers regarded the entire stay in hospital as one event; they did not differentiate between wards or ward staff in the delivery, maternity or neonatal wards. All mothers in the study had, therefore, also experienced part care for shorter or longer periods when separated from their baby, being then later reunited in co-care. CONCLUSION: This study can be used as a basis for discussion on more individualised care through co-operation and organisation between delivery, maternity and neonatal wards, in order to reduce the amount of time mother and baby are separated. PMID- 15878429 TI - Traditional beliefs about pregnancy and child birth among women from Chiang Mai, Northern Thailand. AB - OBJECTIVE: To examine women's embodied knowledge of pregnancy and birth, women's explanations of precautions during pregnancy and birth and preparations for easy birth and the role of a traditional midwife in a Thai birthing care. DESIGN: In depth interviews relating to traditional and changed beliefs and practices of pregnancy and childbirth with Thai women in Northern Thailand. SETTING: Chiang Mai city and Mae On sub-district in Chiang Mai province, Northern Thailand. PARTICIPANTS: 30 Thai women living in Chiang Mai in Thailand. FINDINGS: The social meaning of childbirth in Thai culture is part of the larger social system, which involves the woman, her family, the community, society and the supernatural world. Traditional beliefs and practices in Thai culture clearly aim to preserve the life and well-being of a new mother and her baby. It seems that traditional childbirth practices have not totally disappeared in northern Thailand, but have gradually diminished. Women's social backgrounds influence traditional beliefs and practices. The traditions are followed by most rural and some urban poor women in Chiang Mai. IMPLICATIONS FOR PRACTICE: The findings of this study may assist health professionals to better understand women from different cultures. It is important to recognise many factors discussed in this paper within the context of Thai lives and traditions. This will prevent misunderstanding and, consequently, encourage more sensitive pregnancy and birthing care for pregnant women. PMID- 15878430 TI - Techniques to reduce perineal pain during spontaneous vaginal delivery and perineal suturing: a UK survey of midwifery practice. AB - OBJECTIVE: To investigate use of pharmacological and non-pharmacological methods of perineal analgesia used by midwives during the second stage of labour and perineal repair in the UK. DESIGN: Postal survey. METHODS: Self-complete questionnaires were sent to Heads of Midwifery in all 219 maternity units in the UK. Information was requested on the number and type of deliveries undertaken in the previous year and on the midwifery procedures used to provide pain relief immediately before delivery and for perineal repair. Details were also sought on local anaesthetics given before episiotomy or perineal repair. FINDINGS: 207 completed questionnaires were returned providing information on 210 maternity units. Midwives reported using a variety of non-pharmacological analgesic methods to control pain at the end of the second stage of labour. Hot packs were used in 70 (33%) maternity units, cold packs in 44 (21%) and perineal massage in 109 (52%). Midwives in 131 (62%) maternity units used injectable local anaesthetics to control perineal pain. All units advocated use of local anaesthetic before episiotomy or perineal repair, but the reported doses used varied widely. CONCLUSIONS: The literature on levels of pain experienced immediately before spontaneous vaginal delivery and during perineal repair is sparse, but what evidence exists suggests that, for some women, these occasions are accompanied by severe pain. Findings from this survey show that there is considerable variation in what midwives provide to control pain. Formal evaluation of the perineal analgesia offered to women during the second stage of labour is urgently required. PMID- 15878431 TI - An action-research study exploring midwives' support needs and the affect of group clinical supervision. AB - OBJECTIVE: To explore community midwives' views and experiences of their support needs in clinical practice, and then to identify how they would wish to receive such support. Further objectives were to redress the imbalance identified by planning and facilitating a model of clinical supervision devised by the participating midwives. DESIGN: A qualitative study using an action-research approach based on collaboration and participation. Action research has the potential to facilitate understanding of, and is able to adapt to, changing situations within clinical practice. Data were collected in three phases using in depth interviews and focus groups. SETTING: A large maternity unit in the north of England, UK. PARTICIPANTS: Eight National Health Service (NHS) community midwives working in the same team. FINDINGS: Recent and ongoing organisational change and increased demands placed on the midwives by their managers were found to be detrimental to the process of clinical supervision and working relationships with their peers and clients. These pressures also inhibited the process of change. The midwives' behaviour and coping strategies revealed an apparent lack of understanding on their part, and that of their midwifery managers, of the regulation of emotion and the amount of energy this generated. Pseudo-cohesion and resistance to change were key defence mechanisms used by the participating midwives. KEY CONCLUSIONS: A large amount of published literature supported the existence of stress and burnout in midwifery, but no research addressed ways of alleviating this situation. Effective facilitation of midwifery support is needed, which can be met through support mechanisms such as clinical supervision. During the process of clinical supervision, strong messages emerged about the necessity to ensure that midwives are prepared educationally for the difficult situations that are brought about through collaborative working. There are also messages about the cultural legacy of NHS midwifery and how this can inhibit autonomous behaviour by midwives. IMPLICATIONS FOR PRACTICE: Developing and increasing self-awareness is still not viewed as being intrinsic to the work of the midwife, and midwives are being asked to undertake a level of work for which they have not been adequately prepared. The bureaucratic pressures of working in a large maternity unit exaggerate this further. In this situation, the system is seen as more important than the midwives. PMID- 15878432 TI - 'Maternity clinic on the net service' and its introduction into practice: experiences of maternity-care professionals. AB - OBJECTIVE: To describe the experiences of maternity-care professionals using an Internet-based network service, called Net Clinic, and the opportunities and obstacles they encountered in Net Clinic while organising their work and developing their know-how. DESIGN: Qualitative information gathered from semi structured thematic interviews. SETTING: Four maternity care units in Eastern Finland (one antenatal ward in a university hospital, one antenatal ward in a central hospital, two maternity clinics in the community). PARTICIPANTS: Five midwives, two public health nurses and three doctors. FINDINGS: All participants considered adequate privacy protection a prerequisite for development. They anticipated that because of their computer skills, young people would be able to use information and communication technology (ICT) as part of maternity-care services naturally. Managerial support, such as allocation of time and equipment, was extremely important during the introductory phase. The participants were divided into three groups based on their experiences of implementing Net Clinic: (1) 'doubters' did not believe in their own ICT competence and were afraid of using Net Clinic. They also showed resistance to Net Clinic. Doubters wanted versatile and personal guidance in ICT skills; (2) 'accepters' agreed that progress in ICT would inevitably affect their work. Development was considered difficult without awareness of the benefits of ICT, and Net Clinic was expected to increase their workload. Accepters preferred to have one-to-one ICT training; (3) 'future confidents' saw ICT as a useful tool for developing maternity care. They recognised the opportunities implicit in network collaboration and wanted versatile user training in ICT. They wanted successful network service models and were eager to develop them. IMPLICATIONS FOR THE FUTURE: Families will increase their knowledge levels, as those who will be future parents are learning to use public network services at school. The increasing diversity of family structure will be a challenge to maternity care. Changes in maternity services can be accomplished with new e-Working models developed through diversified co-operation and with managerial support. The crucial question will be the attitude of maternity-care professionals towards Internet-based network service in their work. PMID- 15878433 TI - Writing for publication in this refereed journal. AB - Acceptance of a paper for publication sometimes appears to be a mountain that has to be climbed. For some it has the height of Everest but for others it is a stroll up a hill. In writing this paper I hope that readers will be able to have an easier job in writing a paper that will be accepted in this journal and that in return the editor's task will be a little easier. PMID- 15878434 TI - Calpain and N-methyl-d-aspartate (NMDA)-induced excitotoxicity in rat retinas. AB - Calpain-mediated proteolysis has been implicated as a major process in neuronal cell death in both acute insults and the chronic neurodegenerative disorders in the central nerves system. However, activation of calpain also plays a protective function in the early phase of excitotoxic neuronal death. The exact role of calpains in neuronal death and recovery after exposure to N-methyl-D-aspartate (NMDA) is not clearly known. The purpose of present study was to examine the involvement of mu- and m-calpain in NMDA-induced excitotoxicity in the adult rat retina. Increased immunoreactivity of mu-calpain was noted in RGC layer cells and in the inner nuclear layer with maximal expression at 12 h after NMDA injection. This was further confirmed with Western blotting. TdT-mediated biotin-dUTP nick end labeling (TUNEL) positive cells in the inner retina co-localized with moderate or intense mu-calpain immunoreactivity. In contrast, there was no remarkable change in m-calpain immunoreactivity at any time point after NMDA injection. Simultaneous injection of 2 nmol of a calpain inhibitor (calpain inhibitor II) significantly reduced the number of TUNEL-positive cells in the inner retina at 18 h after NMDA injection and preserved RGC-like cells counted at 7 days after injection. The results of this study showed that mu-calpain may be involved in mediating NMDA-induced excitotoxicity in the rat retina and calpain inhibitors may play a therapeutic role in NMDA related disease. PMID- 15878435 TI - A real-time PCR assay for the detection of Neisseria gonorrhoeae in genital and extragenital specimens. AB - A Neisseria gonorrhoeae LightCycler (NGpapLC) assay targeting the porA pseudogene was compared with bacterial culture for detection of N. gonorrhoeae in 636 clinical specimens (216 cervical, 185 urethral, 196 throat, and 39 rectal swab specimens). The specificity of the NGpapLC assay was further investigated by testing a bacterial reference panel comprising several Neisseria species. Overall, 19 (3.0%) specimens were positive and 613 (96.4%) specimens were negative by both methods. Four (0.6%) specimens were positive by the NGpapLC assay only. For the cervical and urethral swabs, the NGpapLC provided 100% sensitivity and 100% specificity compared with bacterial culture. Following discrepant analysis, the clinical sensitivity and specificity of the NGpapLC for throat and rectal swabs was also 100%. For the bacterial panel, only N. gonorrhoeae isolates provided positive results. The results show the NGpapLC assay is suitable for use on a range of clinical specimens and could improve detection of pharyngeal N. gonorrhoeae. PMID- 15878436 TI - Chlamydia pneumoniae and Mycoplasma pneumoniae in young children from China with community-acquired pneumonia. AB - Eighty-five cases community-acquired pneumonia (CAP) in children 5 years or younger, confirmed by chest X-ray, and 185 age-matched control patients with diarrhea or dermatitis from the Outpatient Department at Beijing Children's Hospital were enrolled into this study. Nasopharyngeal swab specimens were obtained from all subjects. Real-time PCR-based fluorescence assays were performed for Chlamydia pneumoniae and Mycoplasma pneumoniae. A nested PCR was also run for C. pneumoniae for comparison of assays. C. pneumoniae was found in 3 (3.5%) of CAP cases and in 4 (2.1%) of controls (P = 0.51). M. pneumoniae was found in 6 (7.1%) of CAP cases and in none of the controls (P = 0.001). The agreement rate of the 2 applied PCR methods used for C. pneumoniae detection was 98.5%. Our study demonstrates that M. pneumoniae may play a significant role in CAP affecting children up to 5 years in China, whereas C. pneumoniae in nasopharyngeal specimens was not associated with CAP in this age group. PMID- 15878437 TI - Increased frequency of non-fumigatus Aspergillus species in amphotericin B- or triazole-pre-exposed cancer patients with positive cultures for aspergilli. AB - Invasive aspergillosis (IA) can occur despite prior prophylactic or empiric use of triazoles or amphotericin B (AMB). Although profound immunosuppression may account for breakthrough IA, resistance of Aspergillus to antifungals may also play a role. To examine this question, we measured the minimal inhibitory concentration of 105 Aspergillus isolates recovered from 105 cancer patients (64 with IA, 41 with Aspergillus colonization) to AMB, itraconazole (ITC), and voriconazole (VRC) using the National Committee for Clinical Laboratory Standards (NCCLS) M38-A microdilution and E-test methods. We also determined the minimal fungicidal concentration (MFC) of these agents and the minimal effective concentration (MEC) of caspofungin (CAS) using standardized methods. We then collected information regarding pre-exposure to AMB or triazoles (fluconazole, ITC, VRC) within 3 months before Aspergillus isolation. Pre-exposure of cancer patients to AMB or triazoles was associated with increased frequency of non fumigatus Aspergillus species. Aspergillus isolates recovered from patients who previously received AMB exhibited higher E-test AMB MICs compared with isolates from patients without prior AMB exposure (P = 0.01). In addition, the AMB MICs by E-test were higher in triazole-pre-exposed patients compared with those not exposed to triazoles (P = 0.001). The ITC and VRC MICs by E-test were not affected by prior AMB or triazole exposure. Finally, neither the AMB, ITC, and VRC MICs and MFCs by NCCLS method nor CAS MECs showed such changes. In conclusion, cancer patients with positive Aspergillus cultures who are pre exposed to AMB or triazoles have high frequency of non-fumigatus Aspergillus species. These Aspergillus isolates were found to be AMB-resistant by the more sensitive E-test method. PMID- 15878438 TI - The use of variable-number tandem-repeat mycobacterial interspersed repetitive unit typing to identify laboratory cross-contamination with Mycobacterium tuberculosis. AB - A retrospective study including 515 Mycobacterium tuberculosis isolates from 215 patients was conducted to investigate possible laboratory contamination with M. tuberculosis over a 1-year period in a university hospital. All cultures underwent variable-number tandem-repeat (VNTR) typing. Cultures suspected of being contaminated in the VNTR analysis and possible sources of contamination underwent mycobacterial interspersed repetitive unit (MIRU) typing further. Overall, 8 (3.7%) cases of 215 patients were considered possible false-positives. Five (2.3%) cultures might be contaminated during initial batching processing, and 1 (0.5%) and 4 (1.9%) cultures of them were further classified as presumed and possible cases, respectively, of cross-contamination on clinical grounds. Three (1.4%) cultures might be contaminated by cultures that had been processed in species identification procedures in the same laminar-flow hood. The 2-step strategy using VNTR and MIRU analyses in combination in this study appears to be a valuable means for the study of false-positive cultures. PMID- 15878439 TI - Initial management of candidemia at an academic medical center: evaluation of the IDSA guidelines. AB - Treatment of candidemia is more complicated because of the changing epidemiology of Candida and introduction of newer antifungal agents. Utilization and benefit of practice guidelines and infectious disease consultation for the management of candidemia has not been previously described in the routine clinical setting. We prospectively studied the impact of the Infectious Disease Society of America (IDSA) guidelines for the management of candidemia and infectious disease consultation on clinical outcomes in 119 patients with candidemia at a tertiary care hospital. Medical records were reviewed to capture data concerning use of antifungal agents, management of central venous catheters, and infectious disease consultation. Initial antifungal therapy was consistent with the IDSA guidelines in 76% of patients. Variation from the guidelines was independently associated with higher mortality (24% versus 57%, P = 0.003). Infectious disease consultation was independently associated with lower mortality (18% versus 39%, P < 0.01). Use of the IDSA guidelines and infectious disease consultation service was found to improve patient outcomes in patients with candidemia at our institution. Further studies should be performed to validate newer guidelines in a clinical setting at other institutions. PMID- 15878440 TI - Isolation of cholesterol-dependent Candida glabrata from clinical specimens. AB - In the past few years, we have detected in the United Kingdom and in the United States several isolates of Candida glabrata that grew poorly unless bile was available. Cholesterol, a component of bile, stimulated equivalent growth of the bile-dependent isolates. The bile-dependent C. glabrata isolates appeared resistant to amphotericin B, but their resistance to fluconazole was unclear. These results demonstrate that occasional isolates of C. glabrata require cholesterol to grow, they may not be detected in specimens set up on standard primary plating media, and they may be difficult to eradicate in patients with antifungal agents directed against ergosterol and its synthetic pathway. PMID- 15878441 TI - Vinyl adhesive tape also effective for direct microscopy diagnosis of chromomycosis, lobomycosis, and paracoccidioidomycosis. AB - Among the existing techniques for direct microscopy diagnosis in mycology, vinyl adhesive tape (VAT) preparations have proved their usefulness for Malassezia spp., Candida spp., and dermatophyte superficial infections. The collection and identification of fungal agents can be performed easily by the VAT technique, as demonstrated in this work, for the setting of some deep-seated cutaneous mycoses, where infectious agents can be observed in the horny layer of the epidermis in transepidermal elimination events. Preliminary results obtained in a series of 19 cases (chromomycosis, 12; lobomycosis, 5; paracoccidioidomycosis, 2) yielded positive results in 17 (89.47%). Failures for proper microscopic diagnosis detection happened in just 2 cases, 1 for chromomycosis and 1 for paracoccidioidomycosis. These results, although encouraging, were obtained from a relatively small patient population sample. Thus, the authors recommend further tests using this direct microscopy technique in a larger number of patients to validate these preliminary results and to explore its sensitivity in cases of small lesions. PMID- 15878442 TI - Pharmacodynamics of an 800-mg dose of telithromycin in patients with community acquired pneumonia caused by extracellular pathogens. AB - The pharmacodynamics of telithromycin, a new ketolide antibacterial, was examined in 115 patients with community-acquired pneumonia (CAP). Patients received telithromycin 800 mg qd for 7-10 days. Pharmacokinetic parameters were determined, and exposure was linked to microbiological outcome using logistic regression analysis. A breakpoint for increased probability of microbiological eradication was developed and was found to be the ratio of area under the concentration-time curve (AUC) to minimum inhibitory concentration (MIC) of 3.375. The final logistic regression model of microbiological outcome included body weight and AUC/MIC ratio breakpoint. This model was found in analyses of the entire population and when Streptococcus pneumoniae and Haemophilus influenzae were examined separately. The AUC/MIC ratio target attainment rate is expected to be >99.9% for S. pneumoniae and Moraxella catarrhalis and 93.1% for H. influenzae. This study demonstrated a relationship between telithromycin drug exposure and microbiological outcome. Telithromycin is expected to achieve the drug exposure breakpoint for the majority of isolates causing CAP. PMID- 15878443 TI - Trends in linezolid susceptibility patterns in 2002: report from the worldwide Zyvox Annual Appraisal of Potency and Spectrum Program. AB - Oxazolidinones have become reliable clinical and candidate antimicrobial agents to be utilized for infections caused by multidrug-resistant Gram-positive cocci, especially vancomycin-resistant enterococci and methicillin-resistant staphylococci. However, mutational resistance of the ribosomal target has been described for several species. Longitudinal surveillance remains necessary to monitor for this evolving linezolid resistance pattern. A survey of linezolid and several comparison Gram-positive focused agents was initiated in 2002 (7971 strains, >99.0% compliance) for 54 participating sites in the United States, Canada, Europe (6 nations), Latin America (2 nations), and the Asia Pacific (2 nations). The 5 and 25 sites in Canada and the United States, respectively, submitted 200 strains each to a central laboratory for organism identification/confirmation and reference MIC processing. The 10 remaining nations had 200 strain samples from 1 to 4 separate institutions. Linezolid resistance (MIC >/= 8 microg/mL) was confirmed by alternative susceptibility testing methods (Etest, AB BIO Disk, Solna, Sweden; disk diffusion method) and target mutation characterization by PCR and sequence analysis. Linezolid activity against the 6 major organism groups did not vary between geographic areas. A total of 98.1% of linezolid MIC values were between 0.5 and 2 microg/mL, and only 0.5% of results were at 4 microg/mL, which included 32 Staphylococcus aureus (0.9%) and 5 (0.5%) enterococcal isolates. Linezolid resistance was detected in only 4 isolates (0.05%): 1 each Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecium, and a viridans group Streptococcus. All 4 isolates had a G2576U mutation in the 23S rRNA target. Linezolid activity as outlined by these Zyvox Annual Appraisal of Potency and Spectrum (ZAAPS) Program results demonstrate sustained, near complete activity against contemporary Gram positive isolates on 4 monitored continents and in centers utilizing oxazolidinones. Rare linezolid-resistant strains were identified in the United States only (0.05% resistance overall). PMID- 15878444 TI - Penicillin resistance and serotypes/serogroups of Streptococcus pneumoniae in nasopharyngeal carrier children younger than 2 years in Lima, Peru. AB - The purpose of this study was to determine the carriage rate, susceptibility pattern, and serotype distribution of Streptococcus pneumoniae in the nasopharynx of children younger than 2 years old in Lima, Peru. A total of 666 children were evaluated during 3 periods, 1997, 2001, and 2003. The overall pneumococcal carrier rate was 41%. Reduced susceptibility to penicillin was found in 5% (4/75) of isolates in 1997, 20% (15/75) in 2001, and 37% (40/109) in 2003. Reduced susceptibility to ceftriaxone was found in 12% of isolates in 2003. Serogroups 6, 19, 23, 15, and 14 accounted for 68% of all the isolates and for 81% of the penicillin-nonsusceptible strains. Only 65% of the isolated strains had serogroups found in the 7-valent conjugate pneumococcal vaccine. This highlights the importance of regional surveillance studies for effective vaccine strategies and treatment protocols. PMID- 15878445 TI - Interest of postmortem-collected specimens in the diagnosis of fulminant meningococcal sepsis. AB - We reported the case of a child who died of purpura fulminans. The diagnosis of Neisseria meningitidis serogroup C could be assessed using postmortem specimens collected up to 10 h after death. We were able to identify the bacteria by culture and/or PCR on samples without having autopsy performed. Soluble antigens were also detected in serum. PMID- 15878446 TI - Reproducibility assessment of tigecycline MIC results by broth microdilution methods using commercially prepared dry-form panels. AB - We assessed the reproducibility of tigecycline and piperacillin/tazobactam MIC results tested by broth microdilution methods on commercially prepared dry-format panels. Fifteen bacterial isolates were tested 3 times daily for 3 days for a total of 9 replicate results per strain. The within-day and between-days reproducibility was 100% for tigecycline at +/-1 log(2) dilution step and >98% for piperacillin/tazobactam at +/-1 log(2) dilution step. Identical MIC results were observed for 84.4% and 78.7% of tests for same-day replicates for tigecycline and piperacillin/tazobactam, respectively. These results demonstrate the excellent reproducibility of in vitro susceptibility tests for both tigecycline and piperacillin/tazobactam using a commercially prepared dry-form broth microdilution MIC panel. PMID- 15878447 TI - Comparative activity of doripenem and three other carbapenems tested against Gram negative bacilli with various beta-lactamase resistance mechanisms. AB - Doripenem (formerly S-4661), a novel carbapenem antimicrobial, was compared with ertapenem, imipenem, and meropenem using reference broth microdilution test methods against wild-type and various resistant microbial subsets (380 strains). Doripenem and meropenem were consistently more potent than ertapenem or imipenem when tested against Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter spp. Ertapenem exhibited minimum inhibitory concentration (MIC) elevations for some isolates producing AmpC and extended-spectrum beta-lactamases, in contrast to greater enzyme stability for doripenem and other carbapenems tested. Multiple beta-lactamase (TEM, SHV, CTX-M, OXA, CMY types)-producing Escherichia coli had doripenem MIC values at or = 0.84 and P < 0.0001) between 6MTD and 24 h-activity, the slope of which was 2.5-fold greater in the training group (P < 0.01). Similar relationships emerged for 6MWD. There was no association between baseline 6MTD, FEV1 or BMI and any of the other measures. These data suggest that daily activity did not markedly vary with exercise capacity under baseline conditions. Participation in a training program increased activity significantly stronger than predicted from the gain in exercise capacity. This underlines the importance of non physiological, patient-centered factors associated with training in COPD. PMID- 15878488 TI - Sleep-related breathing disorders in obese patients presenting with acute respiratory failure. AB - INTRODUCTION: The study was conducted to assess the clinical and polysomnographic characteristics of patients with sleep-related breathing disorders who presented to the intensive care unit (ICU) with acute respiratory failure and the practicability of performing polysomnography for such patients. MATERIAL AND METHODS: We analyzed clinical presentation, cause of admission to the ICU, ICU course and outcome of 11 subjects with acute respiratory failure who were diagnosed to have sleep disordered breathing based on polysomnography between October 1999 and January 2003. Subjects were compared to 11 patients with obstructive sleep apnea syndrome matched to each subject using body mass index, age and apnea hypopnea index measured at the time of diagnosis (matched comparison group). Repeated arterial blood gases and polysomnography were done for 8 subjects compliant to treatment 6-8 months after discharge from ICU. RESULTS: The reason for ICU admission for all subjects was hypercapnic respiratory failure. pH and daytime PaO2 were significantly lower in studied subjects compared to the matched comparison group while awake daytime PaCO2 was significantly higher. Subjects had frequent episodes of hypoventilation. Follow up arterial blood gases and polysomnography 6-8 months after treatment (non invasive ventilation) in compliant subjects showed significant improvement in all blood gases parameters. CONCLUSIONS: Early polysomnography (or portable cardio respiratory monitoring) allows accurate diagnosis and institution of the appropriate ventilation method Further studies should assess the evolution of respiratory drive in patients with sleep disordered breathing and hypercapnia under therapy (non-invasive ventilation). PMID- 15878489 TI - Hypertonicity of the challenge solution may increase the diagnostic accuracy of histamine challenge. AB - There is significant overlap in the responsiveness to direct airway challenges, such as the histamine challenge, between asthmatic and non-asthmatic subjects, which decreases their accuracy in the diagnosis of asthma. To minimise this overlap, a new test, hypertonic histamine challenge, was developed. Fifteen healthy subjects, 16 subjects with steroid-naive asthma, and 16 asthmatic subjects undergoing inhaled corticosteroid treatment underwent inhalation challenges with hypertonic saline, isotonic histamine, and hypertonic histamine, using an ultrasonic nebuliser and 2-min tidal breathing method. The increase in histamine solution tonicity decreased the histamine PC20 values only in the steroid-naive asthmatic subjects (1.1 (0.5-2.7) vs. 0.5 (0.2-1.2) mg/ml, P = 0.047). Using 1mg/ml as the cut-off value, the sensitivity, specificity, and accuracy of the hypertonic histamine challenge to detect steroid-naive asthma was 81%, 100%, and 90%. The respective values for the isotonic histamine challenge were 56%, 100%, and 77%. Furthermore, there was a statistically significant difference in the hypertonic histamine PC20 between steroid-naive and steroid treated asthmatic subjects, which could not be detected in the isotonic histamine PC20. The hypertonic histamine PC20 was highly repeatable, with a single determination 95% range of +/-1.35 doubling concentrations. The hypertonic histamine challenge was safe but provoked more cough and throat irritation than the other two challenges. In conclusion, compared with a conventional, isotonic histamine challenge, hypertonic histamine challenge may be more accurate in the diagnosis of asthma and also, more capable to detect the effects of inhaled corticosteroid treatment. PMID- 15878490 TI - The relationship between the quality of prescribing and practice appointment rates with asthma management data in those admitted to hospital due to an acute exacerbation. AB - Specific targeting of patients with a previous asthma hospitalisation could be more focused if predictors could be identified. This study was an observational retrospective analysis using ridge and linear multivariate regression analysis. Patient asthma management data were extracted from the hospital and general practice notes of those that had been admitted with an acute exacerbation of their asthma over a 5-year period. From the prescribing data, the annual doses of preventer (P) and reliever (R) medication were converted to defined daily doses then divided to give a P:R ratio. Preliminary statistical analysis was used to identify any association between either the P:R ratio or for the number of general practitioner (GP) practice appointments (PA) and their asthma management data. Multivariate regression analysis was applied to the P:R ratio and to PA to determine a model between each of these and asthma management data/events. GPs gave consent to access the data of 115 (out of 440) asthmatics, age >5 years, admitted to a district general hospital for asthma exacerbations between 1994 and 1998. The multivariate analysis revealed that PA was associated with oral prednisolone rescue courses (PRCs) and age whilst the P:R ratio was associated to PRCs and more reliever usage but not preventers. Patients with low preventer usage with respect to their reliever medication should be targeted for medication review as these were the patients prescribed more prednisolone courses and their increased PAs reflect this. This could decrease visits to the doctor and acute exacerbations. PMID- 15878491 TI - Sputum color as a marker of acute bacterial exacerbations of chronic obstructive pulmonary disease. AB - We analyzed 795 sputa from 315 patients (233 males, mean age 69.3+/-8.8 years, mean number of exacerbations 2.52/patient) with acute exacerbations of moderate to-severe chronic obstructive pulmonary disease (COPD) (mean steady-state FEV1 42.5+/-7.8% of predicted). 581/795 sputa were considered adequate. Sputum was analyzed by a quali-quantitative colorimetric scale allowing both color distinction and color degree of intensity. Quantitative culture was then performed (threshold: >10(6)CFU/mL). Samples were distinguished in mucoid (145) and purulent (436) sputa. Absence of bacterial growth was observed in 22% and 5% of mucoid and purulent sputa, respectively. Among mucoid sputa, Gram positive bacterial growth occurred more commonly compared to Gram negative and Pseudomonas aeruginosa/Enterobacteriaceae (56%, 24%, 20%, respectively). In purulent sputa, Gram positives were found in 38% of cases, Gram negatives in 38%, and P. aeruginosa/Enterobacteriaceae in 24%. We evaluated whether functional impairment (FEV1) orientates as to the infectious etiology of exacerbations. Significant differences were observed in the distribution of pathogens. Gram negative and P. aeruginosa/Enterobacteriaceae were isolated more frequently in the sputum when FEV1 was <35%. Our study indicates that purulent sputum is strongly associated with bacterial growth in COPD exacerbations. Deepening sputum color (from yellowish to brownish) was associated with increased yield of Gram negative and P. aeruginosa/Enterobacteriaceae. PMID- 15878492 TI - Effect of inhalation of thermal water on airway inflammation in chronic obstructive pulmonary disease. AB - Thermal water inhalations have been traditionally used in the treatment of upper and lower chronic airway diseases. However, the benefit and the mechanism of this treatment have not been properly assessed. To determine whether inhaled salt bromide-iodine thermal water improves lung function, quality of life and airway inflammation, 39 patients with chronic obstructive pulmonary disease (COPD) were randomly assigned to receive 2-weeks inhalation treatment with thermal water (active, no. = 20) or normal saline (control, no. = 19) in single blind. Lung volumes were measured, Saint George's respiratory questionnaire (SGRQ) was administered and induced sputum was performed before and after treatment. No changes in pre- and post-salbutamol lung volumes was observed after inhalation treatment in both groups. SGRQ score showed a significant improvement in active group compared with control group at the end of the trial. The concentration of total cells in induced sputum increased significantly in both active (P < 0.05) and control groups (P < 0.05). Inhalation of thermal water induced a small but significant decrease in percentages of sputum neutrophils (P < 0.01) and a parallel increase in macrophages (P < 0.01). In contrast, normal saline inhalation was not associated with changes in differential sputum cell counts. In conclusion, treatment with inhaled salt-bromide-iodine thermal water in COPD is associated with a reduced proportion of neutrophils in induced sputum suggesting that thermal water may have a mild anti-inflammatory effect on the airways. However, the short-term improvement in some components health-related quality of life was not related with changes in lung function or with the degree of airway inflammation. PMID- 15878493 TI - The Medical Research Council dyspnea scale in the estimation of disease severity in idiopathic pulmonary fibrosis. AB - BACKGROUND: Medical Research Council (MRC) chronic dyspnea scale, used for the estimation of disability due to dyspnea, may serve as a simple index of disease severity and extent in patients with idiopathic pulmonary fibrosis (IPF). However, its relationship with other commonly used measures has not been evaluated. METHODS: The association of MRC chronic dyspnea scale with lung function indices and high-resolution computerized tomography (HRCT) scores such as the total interstitial disease score (TIDs) and the fibrosis score (Fs) was examined in 26 untreated patients with IPF sequentially recruited over a period of 3 years. The aim of this observational study was to explore the relationship between dyspnea, impairment of lung function and CT estimation of disease severity in patients with IPF. RESULTS: The MRC dyspnea score was significantly associated with FVC, FEV1, TLC, DLCO, PaO2, and PaCO2 and with both HRCT scores. In multiple regression analysis only the FVC (OR = 0.85, 95% CI = 0.75-0.95, P = 0.004) and PaCO2 (OR = 0.69, 95% CI = 0.50-0.95, P = 0.02) correlated with dyspnea. Furthermore, both TIDs and Fs were negatively associated with FVC, FEV1, TLC and PaO2. In multiple regression analysis only the FVC correlated with both TIDs (r2 = 0.57, P = 0.0001) and Fs (r2 = 0.46, P = 0.0005). CONCLUSIONS: These observations suggest that the MRC dyspnea scale could offer useful information about the estimation of severity in patients with IPF. Furthermore among functional indices the FVC seems to be the best estimator of disease severity and extent. PMID- 15878494 TI - Alcohol-induced upper airway symptoms: prevalence and co-morbidity. AB - Little is known about effects of alcohol intake on the upper, nasal airways. The present aim was to examine the prevalence of alcohol-induced nasal symptoms (ANS) and to explore associations between ANS and other respiratory diseases. A postal questionnaire focused on respiratory diseases and symptoms was sent to 11,933 randomly selected adult individuals. Subjects with ANS, n = 316 (3.4%) received a second questionnaire focusing on this condition. Nine thousand three hundred and sixteen (78%) subjects answered the first and 228 (72%) the second questionnaire. Two-thirds of the subjects with ANS were women. Red wine and white wine were the most frequent triggers of ANS, reported by 83% and 31% of the subjects, respectively. Nasal blockage was the most prominent symptom, but also sneezing, nasal discharge, as well as lower airway symptoms occurred after intake of alcoholic drinks. Self-reported physician's diagnoses of asthma, chronic bronchitis/emphysema, chronic obstructive pulmonary disease (COPD), as well as allergic rhinitis were more common in subjects with ANS compared with the general population (P < 0.001 for all comparisons). In conclusion, ANS are common and are about twice as frequent in women than in men. ANS seem to be associated with important respiratory diseases such as asthma, chronic bronchitis, COPD, and allergic rhinitis. PMID- 15878495 TI - Improvement of asthma control with beclomethasone extrafine aerosol compared to fluticasone and budesonide. AB - Qvar Autohaler efficacy on asthma control, assessed with E. Juniper asthma control questionnaire (ACQ), was compared with fluticasone and budesonide. An open randomized study, stratified (2:1) on the intake of long-acting beta2 mimetics (LAbeta2), was performed in patients with moderate to severe poorly controlled asthma (defined by at least one nocturnal discomfort in the last 5 days or a mean of 2 puffs of short-acting beta2-mimetics in the last 7 days or exercise dyspnea) despite treatment with beclomethasone < or = 1000 microg/day (or equivalent). 460 patients received Qvar Autohaler 800 microg/day (n = 149), fluticasone Diskus 1000 microg/day (n = 149) or budesonide Turbuhaler 1600 microg/day (n = 162) during 12 weeks. Asthma control improved in all groups, with no difference between groups. For patients treated with LAbeta2 (n = 286) a significantly greater improvement of the ACQ score was obtained with Qvar Autohaler versus fluticasone (1.0 +/- 1.0 vs. 0.6 +/- 0.9; P = 0.019), but not versus budesonide (0.9 +/- 0.9). Pulmonary function test improvements were similar in the 3 groups. The significant improvement in asthma control in patients receiving LAbeta2 suggests potential advantages for extrafine aerosols as part of anti-inflammatory treatment optimization. PMID- 15878496 TI - Benefit from anti-inflammatory treatment during clinical remission of atopic asthma. AB - STUDY OBJECTIVES: Subjects with atopic asthma often experience a disappearance of symptoms around puberty. However, airway inflammation and remodeling may persist. It is unknown whether those findings warrant prolonged anti-inflammatory treatment despite the absence of symptoms. In this study, we investigated whether a short course of combined anti-inflammatory treatment would, also in this specific patient population, diminish airway inflammation and/or remodeling. DESIGN: A double-blind, randomized placebo-controlled trial was conducted in 28 asymptomatic subjects with a history of atopic asthma, with established bronchial hyperresponsiveness to methacholine (MCh) as non-invasive indicator of ongoing airway pathology. INTERVENTIONS: Intervention consisted of the salmeterol/fluticasone propionate combination (SFC) product (50/250 microg bid via the Diskus inhaler) or placebo for 3 months. MEASUREMENTS: The change in lung function (FEV1), bronchial response to MCh and adenosine monophosphate (AMP), the fraction of nitric oxide in exhaled air (FENO) and quality of life (QOL) scores were measured. Also, bronchial biopsies were taken and cryo sections immunostained for eosinophils (major basic protein, MBP) and mast cells (tryptase and chymase) before and after treatment. The change in reticular basement membrane (RBM) thickness, one of the parameters of airway remodeling, was also determined. RESULTS: SFC treatment improved hyperresponsiveness to MCh (P = 0.014) as well as AMP (P = 0.011), and reduced FENO (P < 0.001) significantly as compared with placebo. Lung function tended to improve (NS). Furthermore, SFC treatment reduced tryptase in the subepithelium of bronchial biopsy specimens (P = 0.01), and slightly reduced RBM thickness (P = 0.05). However, eosinophils in (sub)epithelium were not significantly affected; neither were chymase levels, blood eosinophils or QOL scores. CONCLUSIONS: We found that 3 months of treatment with fluticasone propionate and salmeterol reduced airway hyperresponsiveness, FENO and tryptase density in the airway mucosa as markers of airway inflammation. MBP density in the airway mucosa and QOL were, however, unchanged. The clinical relevance of these findings, especially with respect to the long-term outcome, has not been determined yet. PMID- 15878497 TI - Monitoring of urine trans, trans-muconic acid level among smokers and non smokers. AB - Smoking has contributed to various neurological, cardiovascular, and pulmonary diseases. According to carcinogens found in cigarette smog, benzene is one of the important carcinogenic compounds. The urinary trans, trans-muconic acid (ttMA) levels among a sample of 10 Thai smokers and 35 Thai non-smokers were investigated, compared and reported. The average urinary ttMA level in smokers and non-smokers were 2.19 +/- 2.32 and 0.24 +/- 0.33 mg/gCr, respectively. A significant higher urinary ttMA level among the smokers was observed (P < 0.05). Since the higher urinary ttMA indicates the higher occult risk for cancer, the usage of urinary ttMA is recommended as a monitoring tool to follow up benzene exposure in the smokers. PMID- 15878498 TI - Desensitization of nicotine acetylcholine receptors: modulation by kinase activation and phosphatase inhibition. AB - The desensitization of alpha-bungarotoxin-insensitive native neuronal nicotinic receptors was studied in rat cortical cell cultures using the patch clamp technique. Thirty-minute perfusions of nicotine reduced currents evoked by short test pulses of 300 microM acetylcholine over a range of 3 to 300 nM, with an IC50 of 51 nM. The time course of desensitization onset was fit by a biexponential function consisting of a fast time constant of about 1 min and a slower component of 6-10 min. The desensitization recovery process was also biexponential and was dominated by a slow time constant of 12-20 min, as well as a minor component of about 1 min. The intracellular dialysis of either the protein kinase C activator phorbol-12-myristate-13 acetate or the phosphatase inhibitor cyclosporin A accelerated the desensitization recovery rate by 2-fold. The data imply that endogenous cortical nicotinic receptor channels may enter one of two desensitization states. The first state (D1) is characterized by rapid entry and recovery, whereas transitions into and out of the second state (D2) occur at slower rates. The D2 receptor state may arise by a sequential transition from the D1 conformation. Protein kinase C activation or phosphatase 2B inhibition may favor the D1 receptor state over that of D2 to promote faster overall rates of desensitization recovery. PMID- 15878499 TI - Effects of betahistine on the spatiotemporal response properties of vestibulospinal neurons to labyrinthine volleys. AB - Betahistine, a drug used in the treatment of vestibular disorders, speeds-up the recovery from hemilabyrinthectomy in experimental animals, likely through the activation of histamine receptors. In order to better understand the mechanism of action of this drug we investigated, in adult, urethane anesthetized rats, whether betahistine modifies the spatial (directional) and temporal response properties of vestibular nuclear neurons to the labyrinthine input, as well as the convergence of different labyrinthine signals on single units. Extracellular single-unit activity was recorded from the caudal, spinal-projecting region of the vestibular nuclei during tilt of the animal, before and after i.p. injection of betahistine. The two orthogonal directions of maximal and minimal response to tilt, as well as the corresponding gains were determined for each neuron. Betahistine reduced the maximal response gain of units showing larger basal values of this parameter and increased it in neurons with smaller basal values, while the minimal response gain was on the average raised. These changes led to a significant decrease in the spatial specificity of the neurons, suggesting that betahistine affects the process of spatiotemporal convergence on vestibular units, likely through a rearrangement of the various inputs. This could be related to the effect of the drug on vestibular compensation. PMID- 15878500 TI - Synthesis and in vitro examination of [124I]-, [125I]- and [131I]-2-(4 iodophenylamino) pyrido[2,3-d]pyrimidin-7-one radiolabeled Abl kinase inhibitors. AB - The pyridopyrimidinones are a potent class of inhibitors of c-Abl kinase and Bcr Abl kinase, the causative fusion protein in chronic myelogenous leukemia and Src family kinases. A novel method for routine, high-yield no-carrier-added synthesis of [(124)I]-, [(125)I]- and [(131)I]-6-(2,6-dichlorophenyl)-2-(4-iodophenylamino) 8-methyl-8H-pyrido[2,3-d]pyrimidin-7-one has been developed. The 4' trimethylstannyl- or 4'-tri-n-butylstannyl-pyridopyrimidinone precursors were prepared from the aryl bromide via a palladium-mediated coupling with hexaalkylditin (dioxane/microwave irradiation/10 min at 160 degrees C). The radioiodination of 4'-stannylpyridopyrimidinones was found to optimally occur via an iododestannylation with Na(124)I, Na(125)I or Na(131)I in the presence of an oxidant [30% H(2)O(2)/HOAc (1:3)/10 min] in 79-87% radiochemical yield with >99% radiochemical purity. The total radiosynthesis time was 30 min. The 4 iodophenylpyridopyrimidinone 2 inhibited recombinant Abl kinase activity with an IC(50) of 2.0 nM. Cell proliferation of K562 and A431 cells was inhibited with an IC(50) of 2.0 and 20 nM, respectively. Rapid cellular uptake and equilibrium were observed within 10-15 min using [(131)I]-4-iodophenylpyridopyrimidinone 6c in K562 and A431 cells and demonstrated a 2.8-fold uptake selectivity for the Bcr Abl-expressing K562 cells at 60 min. These results suggest that pyridopyrimidinone radiotracers may be useful in imaging Abl-, Bcr-Abl- or Src expressing malignancies. PMID- 15878501 TI - Synthesis, 11C labeling and biological properties of derivatives of the tyrphostin AG957. AB - Four analogues of AG957, a known inhibitor of the tyrosine kinase p210(bcr-abl), have been synthesized and tested for their growth inhibitory effect against the BCR/ABL-positive FDrv210C cells as well as the epidermal growth factor (EGF) receptor-positive Baf/ERX cells. All compounds that can undergo oxidation to the corresponding quinone demonstrated inhibition of FDrv210C cells and Baf/ERX cells. Compounds that cannot become oxidized showed significantly less inhibition of BCR/ABL- or EGF receptor-mediated cell proliferation. The (11)C-labeled compounds were prepared by labeling 4-aminobenzoic acid using [(11)C]CH(3)I, which afforded the corresponding (11)C-labeled methyl ester in excellent yields. Subsequent condensation of the amino group with an appropriately substituted hydroxy benzaldehyde formed the respective Schiff base. Reduction of this compound with NaBH(3)CN gave the (11)C-labeled inhibitors in an overall radiochemical yield of 17.3+/-2.1% (n=3; not decay corrected) and an average specific radioactivity of 40 GBq/micromol (1.1 Ci/micromol) at the end of synthesis. The total synthesis time from EOB including HPLC purification and formulation was 45 min. PMID- 15878502 TI - Synthesis and biological evaluation of (E)-3-styrylpyridine derivatives as amyloid imaging agents for Alzheimer's disease. AB - A new series of (E)-3-styrylpyridine derivatives as potential diagnostic imaging agents targeting amyloid plaques in Alzheimer's disease (AD) were synthesized and examined. When in vitro binding studies using AD brain homogenates were carried out with a series of styrylpyridine derivatives, (E)-2-Bromo-5-(4 dimethylaminostyryl)pyridine (7) with a dimethylamino group showed the highest binding affinity. Compound 7 intensely stained neuritic and diffused plaques and cerebrovascular amyloids on postmortem AD brain sections. (E)-2-Iodo-5-(4 dimethylaminostyryl)pyridine, the iodo derivative of compound 7, also stained senile plaques in human AD sections. The radioiodinated ligand [125I] was successfully prepared through an iododestannylation reaction from the corresponding tributyltin derivatives using hydrogen peroxide as the oxidant in high yields and with high radiochemical purity. A biodistribution study in normal mice after an intravenous injection of [125I] displayed high brain uptake and fast washout. Taken together, the data suggest that the new radio tracer, [125I], may be useful as a radioiodinated imaging agent for mapping A beta plaques in the brains of patients with AD. PMID- 15878503 TI - Imaging beta-amyloid fibrils in Alzheimer's disease: a critical analysis through simulation of amyloid fibril polymerization. AB - The polymerization of beta-amyloid (A beta) peptides into fibrillary plaques is implicated, in part, in the pathogenesis of Alzheimer's disease. A beta molecular imaging probes (A beta-MIPs) have been introduced in an effort to quantify amyloid burden or load, in subjects afflicted with AD by invoking the classic PET receptor model for the quantitation of neuronal receptor density. In this communication, we explore conceptual differences between imaging the density of amyloid fibril polymers and neuronal receptors. We formulate a mathematical model for the polymerization of A beta with parameters that are mapped to biological modulators of fibrillogenesis and introduce a universal measure for amyloid load to accommodate various interactions of A beta-MIPs with fibrils. Subsequently, we hypothesize four A beta-MIPs and utilize the fibrillogenesis model to simulate PET tissue time activity curves (TACs). Given the unique nature of polymer growth and resulting PET TAC, the four probes report differing amyloid burdens for a given brain pathology, thus complicating the interpretation of PET images. In addition, we introduce the notion of an MIP's resolution, apparent maximal binding site concentration, optimal kinetic topology and its resolving power in characterizing the pathological progression of AD and the effectiveness of drug therapy. The concepts introduced in this work call for a new paradigm that goes beyond the classic parameters B(max) and K(D) to include binding characteristics to polymeric peptide aggregates such as amyloid fibrils, neurofibrillary tangles and prions. PMID- 15878504 TI - A preliminary PET evaluation of the new dopamine D2 receptor agonist [11C]MNPA in cynomolgus monkey. AB - This study describes the preliminary positron emission tomography (PET) evaluation of a dopamine D(2)-like receptor agonist, (R)-2-(11)CH(3)O-N-n propylnorapomorphine ([(11)C]MNPA), as a potential new radioligand for in vivo imaging of the high-affinity state of the dopamine D(2) receptor (D(2)R). MNPA is a selective D(2)-like receptor agonist with a high affinity (K(i)=0.17 nM). [(11)C]MNPA was successfully synthesized by direct O-methylation of (R)-2-hydroxy NPA using [(11)C]methyl iodide and was evaluated in cynomolgus monkeys. This study included baseline PET experiments and a pretreatment study using unlabeled raclopride (1 mg/kg). High uptake of radioactivity was seen in regions known to contain high D(2)R, with a maximum striatum-to-cerebellum ratio of 2.23+/-0.21 at 78 min and a maximum thalamus-to-cerebellum ratio of 1.37+/-0.06 at 72 min. The pretreatment study demonstrated high specific binding to D(2)R by reducing the striatum-to-cerebellum ratio to 1.26 at 78 min. This preliminary study indicates that the dopamine agonist [(11)C]MNPA has potential as an agonist radioligand for the D(2)-like receptor and has potential for examination of the high-affinity state of the D(2)R in human subjects and patients with neuropsychiatric disorders. PMID- 15878505 TI - Candidate PET radioligands for cannabinoid CB1 receptors: [18F]AM5144 and related pyrazole compounds. AB - INTRODUCTION: The mammalian brain contains abundant G protein-coupled cannabinoid CB(1) receptors that respond to Delta(9)-tetrahydrocannabinol, the active ingredient of cannabis. The availability of a positron emission tomography (PET) radioligand would facilitate studies of the addictive and medicinal properties of compounds that bind to this receptor. Among the known classes of ligands for CB(1) receptors, the pyrazoles are attractive targets for radiopharmaceutical development because they are antagonists and are generally less lipophilic than the other classes. METHODS: A convenient high-yield synthesis of N-(4 [(18)F]fluorophenyl)-5-(4-bromophenyl)-1-(2,4-dichlorophenyl)-1H-pyrazole-3 carboxamide (AM5144) was devised by coupling the appropriate pyrazole-3-carboxyl chloride compound with 4-[(18)F]fluoroaniline. The labeled precursor was synthesized from 1-[(18)F]fluoro-4-nitrobenzene in 60% radiochemical yield for 10 min using an improved procedure involving sodium borohydride reduction with cobalt chloride catalysis. The product was purified by HPLC to give a specific activity >400 mCi/micromol and a radiochemical purity >95%, and a PET study was conducted in a baboon. RESULTS: Although the regional uptake of AM5144 in baboon brain was consistent with binding to cannabinoid CB(1) receptors, absolute uptake at <0.003% injected radioactivity per cubic centimeter was lower than the previously reported uptake of the radioiodinated pyrazole AM281. CONCLUSIONS: The relatively poor brain uptake of AM5144 and other pyrazole CB(1) receptor ligands is not surprising because of their high lipophilicity as compared with most brain PET radiotracers. However, for nine pyrazole compounds for which rodent data are available, brain uptake and calculated logP values are not correlated. Thus, high logP values should not preclude evaluation of radiotracers for targets such as the CB(1) receptor that may require very lipophilic ligands. PMID- 15878506 TI - Evaluation of F-18-labeled amino acid derivatives and [18F]FDG as PET probes in a brain tumor-bearing animal model. AB - 2-Deoxy-2-[(18)F]fluoro-d-glucose ([(18)F]FDG) has been extensively used as positron emission tomography (PET) tracer in clinical tumor imaging. This study compared the pharmacokinetics of two (18)F-labeled amino acid derivatives, O-2 [(18)F]fluoroethyl-l-tyrosine (l-[(18)F]FET) and 4-borono-2-[(18)F]fluoro-l phenylalanine-fructose (l-[(18)F]FBPA-Fr), to that of [(18)F]FDG in an animal brain tumor model. METHODS: A self-modified automated PET tracer synthesizer was used to produce no-carrier-added (nca) l-[(18)F]FET. The cellular uptake, biodistribution, autoradiography and microPET imaging of l-[(18)F]FET, l [(18)F]FBPA-Fr and [(18)F]FDG were performed with F98 glioma cell culture and F98 glioma-bearing Fischer344 rats. RESULTS: The radiochemical purity of l-[(18)F]FET was >98% and the radiochemical yield was 50% in average of 16 runs. The uptake of l-[(18)F]FET and l-[(18)F]FBPA-Fr in the F98 glioma cells increased rapidly for the first 5 min and reached a steady-state level after 10 min of incubation, whereas the cellular uptake of [(18)F]FDG kept increasing during the study period. The biodistribution of l-[(18)F]FET, l-[(18)F]FBPA-Fr and [(18)F]FDG in the brain tumors was 1.26+/-0.22, 0.86+/-0.08 and 2.77+/-0.44 %ID/g at 60 min postinjection, respectively, while the tumor-to-normal brain ratios of l [(18)F]FET (3.15) and l-[(18)F]FBPA-Fr (3.44) were higher than that of [(18)F]FDG (1.44). Both microPET images and autoradiograms of l-[(18)F]FET and l-[(18)F]FBPA Fr exhibited remarkable uptake with high contrast in the brain tumor, whereas [(18)F]FDG showed high uptake in the normal brain and gave blurred brain tumor images. CONCLUSION: Both l-[(18)F]FET and l-[(18)F]FBPA-Fr are superior to [(18)F]FDG for the brain tumor imaging as shown in this study with microPET. PMID- 15878507 TI - Synthesis, in vitro binding and biodistribution in B16 melanoma-bearing mice of new iodine-125 spermidine benzamide derivatives. AB - In the course of our investigations aimed at improving the biological characteristics of iodobenzamides for melanoma therapeutic applications, four new derivatives containing a spermidine chain have been prepared and radiolabeled with (125)I. In vitro studies showed that all compounds displayed high affinity for melanin superior to the reference compound BZA, thus validating our experimental approach. In vivo biodistribution was investigated in B16 melanoma bearing mice. All four compounds, particularly benzamide 3, showed accumulation in the tumor, but lower, however, than that of BZA. Moreover, high concentrations of radioactivity in other organs, namely, the liver and lung, demonstrated nonspecific tumoral uptake. In view of these results, compounds 1 2 3 4 do not appear to be suitable radiopharmaceuticals for melanoma radionuclide therapy. PMID- 15878508 TI - Radiosynthesis and evaluation of two novel 123I-labeled 2-methyl-4-nitroimidazole derivatives as potential infection imaging agents. AB - INTRODUCTION: The inflammation- and infection-seeking properties of (131)I labeled ornidazole, a 5-nitroimidazole derivative, have recently been reported. Whole-body images in rabbits showed a more rapid uptake in inflamed areas compared to (67)Ga. In the present study, two novel (123)I-labeled 2-methyl-4 nitroimidazole derivatives were synthesized and their infection-seeking properties compared with those of (67)Ga and (123)I-labeled ornidazole. METHODS: Radiolabeling was carried out by means of iodide-for-tosylate, triflate or halogen exchange. Various methods were utilized in order to synthesize the labeling precursors for the (123)I-labeled novel compounds. Serum stability studies on all of the (123)I-labeled tracers were followed by gamma camera imaging studies on rabbits artificially infected with Escherichia coli bacteria. RESULTS AND CONCLUSIONS: The (123)I-labeled tracers were obtained in moderate to good radiochemical yields (34-80%) and acceptable radiochemical purities (93 99%). In contrast to (123)I-labeled ornidazole, 1-[(1-hydroxy-3-[(123)I]iodoprop 2-yloxy)methyl]-2-methyl-4-nitroimidazole (2) and 1-[(1-[(123)I]iodoprop-2 yloxy)methyl]-2-methyl-4-nitroimidazole (3) showed high serum stability. Compared to noninfected controls, all of the (123)I-labeled tracers showed increased uptake at the area of induced infection after 6 and 24 h, but the uptake was significantly lower than in the case of (67)Ga over the same period. Tracer 3 showed a slightly superior uptake after 6 h than the other (123)I-labeled tracers over the same period. The advantage of the initially slightly faster rate at which nitroimidazole tracers appear to accumulate in the infection area in comparison to (67)Ga might not outweigh the advantage of the eventual higher target to nontarget ratio displayed by (67)Ga. PMID- 15878509 TI - Imaging apoptosis in vivo using 124I-annexin V and PET. AB - Abnormal regulation of apoptosis is an important pathogenic mechanism in many diseases including cancer. Techniques to assess apoptosis in living organisms are limited and, in the case of solid organs, restricted to histological examination of biopsy samples. We investigated the use of (124)I-annexin V, which binds to phosphatidylserine (PS) on the surface of apoptotic cells, as a potential positron emission tomography (PET) radioligand for the noninvasive measurement of apoptosis in vivo. Annexin V and a similar-sized protein, ovalbumin, were directly labelled with (124)I. We report the validation of (124)I-annexin V in vitro and in an animal model of liver apoptosis that has not previously been used to test iodinated annexin V. Also, for the first time, we report metabolite analysis of (124)I-annexin V and the correlation of (124)I-annexin V uptake with apoptotic density (AD). Sixfold more (124)I-annexin V was associated with Jurkat cells after apoptosis induction, indicating that PS binding by annexin V was preserved after iodination. (124)I-ovalbumin did not demonstrate increased uptake in apoptotic cells. In normal BDF-1 mice, the radioligand was rapidly cleared, but some in vivo dehalogenation resulted in the accumulation of activity in the thyroid and stomach content. PET images demonstrated uptake of (124)I-annexin V but not (124)I-ovalbumin in apoptotic liver lesions. In vivo (124)I-annexin V uptake, derived from PET images, correlated with histologically derived AD (r=.86, P<.01). These results demonstrate that (124)I-annexin V is localised to anti-Fas-induced apoptosis, in contrast to (124)I-ovalbumin, which did not show preferential uptake in the apoptotic liver. PMID- 15878510 TI - Functional comparison of annexin V analogues labeled indirectly and directly with iodine-124. AB - We are interested in imaging cell death in vivo using annexin V radiolabeled with (124)I. In this study, [(124)I]4IB-annexin V and [(124)I]4IB-ovalbumin were made using [(124)I]N-hydroxysuccinimidyl-4-iodobenzoate prepared by iododestannylation of N-hydroxysuccinimidyl-4-(tributylstannyl)benzoate. [(124)I]4IB-annexin V binds to phosphatidylserine-coated microtiter plates and apoptotic Jurkat cells and accumulates in hepatic apoptotic lesions in mice treated with anti-Fas antibody, while [(124)I]4IB-ovalbumin does not. In comparison with (124)I-annexin V, [(124)I]4IB-annexin V has a higher rate of binding to phosphatidylserine in vitro, a higher kidney and urine uptake, a lower thyroid and stomach content uptake, greater plasma stability and a lower rate of plasma clearance. Binding of radioactivity to apoptotic cells relative to normal cells in vitro and in vivo appears to be lower for [(124)I]4IB-annexin V than for (124)I-annexin V. PMID- 15878511 TI - Synthesis, enantiomeric resolution, F-18 labeling and biodistribution of reboxetine analogs: promising radioligands for imaging the norepinephrine transporter with positron emission tomography. AB - Racemic and enantiomerically pure ((S,S) and (R,R)) 2-[alpha-(2-(2 [(18)F]fluoroethoxy)phenoxy)benzyl]morpholine ([(18)F]FRB) and its tetradeuterated form [(18)F]FRB-D(4), analogs of the highly selective norepinephrine reuptake inhibitor reboxetine (2-[alpha-(2 ethoxyphenoxy)benzyl]morpholine, RB), have been synthesized for studies of norepinephrine transporter (NET) system with positron emission tomography (PET). The [(18)F]fluorinated precursor, (S,S)/(R,R)-N-tert-butyloxycarbonyl-2-[alpha-(2 hydroxyphenoxy)benzyl]morpholine ((S,S)/(R,R)-N-Boc-desethylRB), was prepared by the N-protection of (S,S)/(R,R)-2-[alpha-(2-hydroxyphenoxy)benzyl]morpholine ((S,S)/(R,R)-desethylRB) with a tert-butyloxycarbonyl (Boc) group followed by enantiomeric resolution with chiral HPLC to provide both (S,S) and (R,R) enantiomers with >99% enantiomeric purity. These compounds were then used for radiosynthesis to prepare enantiomerically pure [(18)F]FRB and [(18)F]FRB-D(4) via the following three-step procedure: (1) formation of 1-bromo-2 [(18)F]fluoroethane ([(18)F]BFE or [(18)F]BFE-D(4)) by nucleophilic displacement of 2-bromoethyl triflate (or D(4) analog) with no-carrier added [(18)F]F(-) in THF; (2) reaction of [(18)F]BFE (or [(18)F]BFE-D(4)) with N-Boc-desethylRB in DMF in the presence of excess base; and (3) deprotection with trifluoroacetic acid. The racemates, (S,S) and (R,R) enantiomers of [(18)F]FRB and [(18)F]FRB-D(4) were obtained in 11-27% (decay corrected to the end of bombardment, EOB) in 120-min synthesis time with a radiochemical purity of >98% and specific activities of 21 48 GBq/micromol (EOB). The results of the whole-body biodistribution studies with (S,S)-[(18)F]FRB-D(4) were similar to those with (S,S)-[(18)F]FRB but showed relatively faster blood clearance and no significant in vivo defluorination. Positron emission tomography studies in baboon brain also showed that (S,S) [(18)F]FRB-D(4) may be a potentially useful ligand for imaging NET with PET. PMID- 15878512 TI - Protein C inhibitor (plasminogen activator inhibitor-3) expression in the CWR22 prostate cancer xenograft. AB - The serine protease inhibitor (serpin) protein C inhibitor (PCI) has been found in the prostate and possibly is a marker to distinguish normal prostate, benign prostatic hyperplasia, and prostate cancer. In this study, we assessed PCI expression in normal, hyperplastic, and malignant prostatic tissues, prostate cancer cell lines, and the CWR22 prostate cancer xenograft model that allowed us to study PCI expression and its regulation in response to androgens. By Northern blot, immunohistochemistry, and in situ hybridization, we found that PCI was expressed in both benign and malignant prostate tissues. Protein C inhibitor was expressed in both androgen-independent (PC-3) and androgen-dependent (LNCaP) prostate cancer cell lines. Furthermore, PCI was detected in all CWR22 tumor samples (androgen dependent, 6 days post-castration, 12 days post-castration followed by 72 h of testosterone treatment, and recurrent CWR22 tumor), although expression of the mature forms of both prostate-specific antigen (PSA) and its homolog, kallikrein 2 (hK2), was clearly androgen-dependent. These results suggest that PCI expression is not regulated by androgens and that PCI is unlikely to be a tumor suppressor gene, but also that PCI may be involved in regulating key serine proteases involved in metastatic prostate disease. PMID- 15878513 TI - Prevention of halo pin complications in post-cranioplasty patients. AB - INTRODUCTION: Distraction Osteogenesis has been successfully implemented in the treatment of maxillary hypoplasia. By using the Rigid External Distraction device (RED) the maxilla can be advanced without the need for bone grafts, providing more stability to the repositioned maxilla. BACKGROUND: The introduction of the RED system in craniofacial surgery has given rise to previously unseen problems. AIM: To provide a set of protocol improvements that might prevent the intracranial pin migration seen at the removal of a RED-II in one patient. CONCLUSIONS: Although the RED device has been shown to achieve good clinical results, there are some disadvantages to the system. There is a high incidence of pin tract infections - leading to loosening of the pins and loss of rigidity. Also, the external ring is prone to traumatic injury. Furthermore, the positioning of the pins can be difficult in thin bone. The improvements used in our protocol might prevent this complication. PMID- 15878514 TI - Definitive surgical correction of the deformity resulting from hemimandibular hyperplasia. AB - INTRODUCTION: The asymmetric deformity of the mandible resulting from the rare condition of hemimandibular hyperplasia has posed a challenge for aesthetic surgical correction. The literature relating to this condition is reviewed and the results of treatment in a series of patients described. MATERIAL AND METHODS: A series of treated patients is reviewed and the results of surgery illustrated and discussed. A maxillary Le Fort I osteotomy is first performed where levelling of the occlusal plane is required. The mandibular surgical technique commences with an extended sagittal ramus osteotomy with dissection of the inferior dental neurovascular bundle completely free of the mandible up to and including the mental foramen. The proximal fragment is then rotated cranially leaving undisturbed the soft tissue attachments at the inferior border, with corresponding bone reduction at the upper border. The inferior border of the distal fragment is then reduced to match the new position of the lower border of the proximal fragment, the neurovascular bundle repositioned, and rigid fixation applied. Finally, a straightening genioplasty is performed. RESULTS: It was possible to overcome the technical and anatomical difficulties associated with correction of this hemimandibular deformity and to achieve a good aesthetic result with acceptable facial symmetry when applying this surgical technique in 3 patients. CONCLUSION: Hemimandibular hyperplasia can be surgically corrected with good aesthetic and functional results and minimal morbidity. PMID- 15878515 TI - Relationship between the risk of mandibular angle fractures and the status of incompletely erupted mandibular third molars. AB - BACKGROUND: Several studies have shown the increased risk of mandibular angle fractures associated with incompletely erupted mandibular third molars. But only a few reports analysed in detail the relationship of the status of eruption of this tooth and this type of fracture. OBJECTIVES: The purpose of this study was to clarify the influence of the eruption status of incompletely erupted mandibular third molars on the incidence of mandibular angle fractures. METHODS: Four hundred and thirty-six mandibular halves in 218 patients with mandibular fractures, aged between 15 and 40 years old, were analysed using panoramic radiographs. RESULTS: The incidence of angle fractures in the mandibular halves with incompletely erupted mandibular third molars was 30.8% and this was statistically significantly higher than that in another group (p<0.0001). Deeply located mandibular third molars clearly showed a higher incidence of this fracture when compared with the adjacent second molar (p<0.0001). CONCLUSION: The results of this investigation showed that incompletely erupted mandibular third molars close to the inferior border of the mandible have a high risk of angle fractures. PMID- 15878516 TI - Relationship between lower wisdom tooth position and cyst development, deep abscess formation and mandibular angle fracture. AB - AIM: There is an ongoing controversy about indications for prophylactic removal of third molars. The purpose of this retrospective study was to clarify the risk of preservation of lower third molars. MATERIAL AND METHOD: In a 5-year period, 316 patients were registered who had received in-patient treatment for deep abscess formation, cyst formation or mandibular angle fracture in relation to lower third molars. A radiological analysis (panoramic radiographs) was performed to determine whether major pathological changes associated with lower wisdom teeth are related to their position. Third molar positions were studied in this in-patient group and in an out-patient group. The latter consisted of 300 consecutive patients with prophylactically removed impacted third molars without any pathology. The relationship between the positions and the different pathological changes associated with impacted lower wisdom teeth was analysed statistically using a new 'position score'. RESULTS: The study revealed that the highest 'position scores' corresponding to a leading aberrant position correlated significantly with cyst formation. Lower scores corresponding to moderately aberrant or slightly irregular position were found with angle fractures, abscess formation, and in the control group as a whole. CONCLUSION: This study indicates that prophylactic third molar surgery for teeth with high and strongly elevated 'position scores' is appropriate in order to prevent cyst formation or mandibular angle fractures in a population at risk for facial trauma. In addition to other factors, 'position score' data could be useful for development of a model for predicting severe complications related to (removal of) impacted lower wisdom teeth. PMID- 15878517 TI - The periauricular transparotid approach for open reduction and internal fixation of condylar fractures. AB - INTRODUCTION: Treatment of fractures of the mandibular condyle fractures varies among centres as there still is no general consensus. The aim of this paper was to determine the safety and efficiency of surgical treatment using a transparotid approach for direct plating. PATIENTS AND METHODS: A prospective clinical study was conducted on 34 patients with 36 fractures of the condyle. All 36 fractures were displaced, and 14 (39%) of them were fracture dislocations. The fractures were treated surgically with a transparotid facelift or retromandibular approach using miniplates and screws for fixation. Patients were carefully followed up and were asked to answer a survey paper 2-39 months postoperatively. RESULTS: Occlusion practically identical to the pretraumatic condition was achieved in 31 out of 33 dentate patients (94%). Postoperative interincisal distance was 30-61 mm (mean 44 mm), 4 patients (12%) had postoperative deflection to the side of injury during mouth opening. Facial symmetry was achieved in all of the patients. Eight out of 36 cases (22%) had a transient weakness of certain ipsilateral facial muscle groups, lasting for 4-8 weeks. In one of these patients, a mild weakness of the upper lip and lower eyelid persisted after 13 months. There were 5 cases of miniplate fractures (14%), all of them in patients in whom 1.7 or thinner miniplates were used. There were 5 cases of salivary fistulae (14%), all of them in patients where the parotid capsule was not closed in a watertight fashion. According to the postoperative survey completed by 32 patients, 30 of them (94%) were very satisfied with the outcome of treatment. CONCLUSION: If conducted properly, the transparotid facelift approach offers a safe and effective approach for direct fixation of condylar fractures. PMID- 15878518 TI - A comparative study of unilateral dislocated mandibular condyle fractures in the rabbit. AB - PURPOSE: The purpose of this study was to clarify the influence of patients age and the degree of dislocation on the healing of mandibular condyle fractures. MATERIAL AND METHODS: Sixty-two Japanese white rabbits (30 adolescent and 30 adult ones) were used. They underwent unilateral condylar neck osteotomy. The proximal fragments were positioned differently. In group I animals, the fragments were reduced into the original position, in group II animals the small fragment was dislocated 55-90 degrees to the ramus. In group III animals the proximal fragment was dislocated 135 degrees to the ramus. Gross and microscopic assessment was performed 4, 6, 8, 12 and 24 weeks postoperatively. RESULTS: In adolescent animals, complete healing of the fracture was observed, irrespective of the degree of dislocation. In the adult animals, there was complete healing of the fractures in groups I and II. In group III animals, there was a decrease of ramus height accompanied by some fibro-cartilaginous changes. CONCLUSION: Even in rabbits condylar deformation has been found following heavily dislocated artificial condylar fractures in adult animals. This result will help to decide for or against surgical treatment of condylar fractures in humans. PMID- 15878519 TI - Hyperostosis as a late sequel of parasymphyseal mandibular fractures in 2 children. AB - BACKGROUND: The potential problem of growth anomalies affecting a mandible following a fracture of a mandibular condyle in childhood is well established. However, there have been no previous reports of this phenomenon affecting other fracture sites in the mandible. PATIENTS: Two patients who had parasymphyseal fractures treated in childhood presented at skeletal maturity with hyperostosis at the fracture site, producing chin asymmetry in their teens. RESULTS: In both cases the hyperostosis produced significant chin asymmetry without disturbance of the occlusion. Both patients were managed with corrective genioplasty. CONCLUSION: These cases reinforce the previous recommendations regarding the need for long-term follow-up of children who sustain facial fractures of the mandible, and that the protocol should be expanded to include parasymphyseal fractures as well as fractures of the condyle. PMID- 15878520 TI - Tumour-associated urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 in normal and neoplastic tissues of patients with squamous cell cancer of the oral cavity - clinical relevance and prognostic value. AB - The central role of the serine protease urokinase-type plasminogen activator (uPA) and its inhibitor, the plasminogen activator-inhibitor-1 (PAI-1), in tumour invasion and metastasis becomes more and more evident. In several studies, uPA and PAI-1 proved to be of prognostic relevance as shown for different types of cancer (e.g. breast, stomach, lung). Elevated antigen levels of uPA and/or PAI-1 predict poor outcome (relapse-free survival) for patients afflicted with cancer. For oral squamous cell carcinomas, however, the prognostic relevance of the tumour-associated proteolytic factors uPA and PAI-1 has still to be evaluated. In the present study, using tissue extracts of 79 oral cancer cases, 58 specimens of normal oral cavity mucosa and of 16 tumour positive lymph nodes taken from the same patients, uPA and PAI-1 antigen were determined by highly sensitive enzyme linked immunosorbent assays (ELISA). A correlation was found between uPA and PAI 1 in tumour tissue, when compared with the normal mucosa of the same oral cavity. Median levels showed significant elevations in cancer tissue and in tumour positive lymph nodes versus normal oral mucosa. In patients with high levels of uPA or PAI-1, there were significantly more tumour relapses. There was no significant correlation between pathological TNM categories, grading, residual tumour category, tumour site and patient age. In summary, tumour uPA/PAI-1 content (as determined by ELISA) appears to be a strong independent prognostic factor for relapse-free survival in squamous cell cancer of the oral cavity. These observations might help to select patients with poor prognosis for additional adjuvant therapy in conjunction with complete surgical resection. PMID- 15878521 TI - Infiltrating basal cell carcinoma of the neck 34 years after irradiation of an haemangioma in early childhood. A case-report. AB - A case of a 34-year-old Caucasian male is presented with a basal cell carcinoma deeply infiltrating the structures of the neck, including skeletal muscles and reaching the parotid gland. Radical surgical removal under meticulous histological control was performed. 18 months postoperatively, the patient is still free of recurrence. Thirty years ago the patient had undergone radiotherapy for an infantile haemangioma of the skin of the neck. The effects of this treatment when given in childhood in the aetiology of a basal cell carcinoma are discussed. PMID- 15878522 TI - Early CT-scan for chronic lacrimal duct symptoms - case report of a malignant melanoma of the lacrimal sac and review of the literature. AB - Malignant melanoma of the lacrimal sac is rare, and only 21 cases have been reported in the world literature. They have an insidious onset and may look like chronic dacryocystitis. In only 6 of these cases were there no recurrences. In most of the survivors, the diagnosis was made early which appears to be the most important prognostic factor. Hence a CT-scan in cases of persistent dacryocystitis is important. Melanoma of the lacrimal duct system is a good example of a condition requiring a multidisciplinary approach between ophthalmologist and maxillofacial surgeon. Any persistently symptomatic lacrimal duct system may need a maxillofacial consultation. PMID- 15878523 TI - Appraisal of a modified medial canthal plication for treating laxity of the medial lower eyelid. AB - PURPOSE: The purpose of this study was to evaluate the efficacy of modified medial canthal tendon plication technique for correcting laxity of the medial end of the lower eyelid. MATERIAL AND METHODS: Eleven patients (9 males and 2 females, 21 eyes), with an age range of 31-80 years, having laxity of the medial end of the lower lid of varying degrees were enrolled in this study. These patients presented with complaints of watering, recurrent redness, photophobia and foreign body sensation. After grading the amount of lower lid laxity, plication was performed. In patients in whom laxity was associated with ectropion, the severity of lower lid ectropion was also graded. In patients with ectropion of Grade II or more, additional procedures for its correction were performed before undertaking plication. The patients were followed up 6 months post-operatively and re-assessed for laxity, recurrence of symptoms and complications of plication. RESULTS: All the lids were evaluated for the amount and extent of laxity of lower lid. Twelve (57%) eyes had Grade I, 9 (43%) had Grade II (and none Grade III) laxity of the lower lid. Twelve eyes had laxity restricted to the medial end and 9 eyes had laxity of the entire length of the lower lid. Some eyes also had an associated ectropion. Among the 21 eyes, 4 eyes (22%) had Grade 1, 11 (61%) had Grade 2, 6 (17%) Grade 3 and none Grade 4 ectropion. In the lids with associated ectropion, additional procedures had been performed. In 1 eye, the medial canthus was anchored to the tendon and this patient had poor apposition of the lower lid to the globe. Hence, in the remaining 20 eyes, the medial canthus was anchored to the orbital periosteum. The mean change in eyelid length was 2.6 mm (measured from medial to lateral canthus) from pre-operative status (33+/-3.4 mm) to the post-operative status (30+/-2.9 mm) this being statistically significant. An overriding of the lower lid was seen in 2 eyes (10%), and a residual gap between the lower lid and globe was noticed in 15 (71%) eyes. Some eyes had uncorrected symptoms including epiphora (3 eyes; 14%), redness (2 eyes; 10%), and photophobia (1 eye; 5%). All were those in which the lateral end was lax. None had a recurrence of foreign body sensation. CONCLUSION: Plication is easy, effective and quick for correcting laxity of the lower lid when restricted predominantly to the medial end. Correct identification of anatomical landmarks and appropriate tension of sutures is vital to achieve proper apposition of the lower lid to the globe. PMID- 15878524 TI - Transient recurrence of pressure regurgitation following successful conventional external dacryocystorhinostomy. AB - PURPOSE: To report a patient with transient recurrence of lacrimal pressure regurgitation following a successful conventional external dacryocystorhinostomy. MATERIAL AND METHODS: A 22-year-old female, who had undergone conventional external dacryocystorhinostomy, presented 3 days later with a recurrence of pressure regurgitation. The patient's symptoms disappeared spontaneously 7 days later. Serial naso-endoscopic examination was used to find the cause. RESULTS: Endoscopic examination showed an oedematous posterior nasal mucosal remnant obstructing the lacrimal drainage pathway. On account of its peculiar location, shape and movements, and the surgically induced tissue oedema, this mucosal remnant misdirected the fluid coming through the canalicular system away from the nasal cavity into the lacrimal pocket. Ten days later, due to healing fibrosis, this flap adhered permanently to the lateral nasal wall and the patient's symptoms disappeared spontaneously. CONCLUSION: Nasal endoscopy is a simple, quick and convenient technique for diagnosing the cause of obstruction in the lacrimal drainage system. The surgeon should aim for large anterior lacrimal and anterior nasal mucosal flaps, and every remnant of posterior flaps should be excised during dacryocystorhinostomy. PMID- 15878525 TI - Failed dental implants - clinical, radiological and bacteriological findings in 17 patients. AB - AIM: The aim of this study was to evaluate the reasons for implant failure in two Oral and Maxillofacial Units in Helsinki. Seventeen patients, who lost 30 implants were included in this study. MATERIAL AND METHODS: The patients and implants were studied clinically, radiologically and microbiologically. RESULTS: Most patients did not have any symptom indicating failure; hence, the failures were noticed mainly by the clinicians when instability of the fixture or of the prosthetic reconstruction became obvious. Radiolucency around the fixture was the most frequent radiological finding. Twenty per cent of the fixtures were located in insufficient bone. Ninety seven per cent of the bacterial cultures were positive, Streptococcus milleri being the most commonly identified aerobic and Fusobacterium nucleatum the most commonly anaerobic bacteria. CONCLUSION: The most critical time for success is immediately after prosthetic loading. Hence, implants should be placed in the optimal position to facilitate prosthetic reconstruction and loading. PMID- 15878526 TI - Multinuclear giant cell formation is enhanced by down-regulation of Wnt signaling in gastric cancer cell line, AGS. AB - AGS cells, which were derived from malignant gastric adenocarcinoma tissue, lack E-cadherin-mediated cell adhesion but have a high level of nuclear beta-catenin, which suggests altered Wnt signal. In addition, approximately 5% of AGS cells form multinuclear giant cells in the routine culture conditions, while taxol treatment causes most AGS cells to become giant cells. The observation of reduced nuclear beta-catenin levels in giant cells induced by taxol treatment prompted us to investigate the relationship between Wnt signaling and giant cell formation. After overnight serum starvation, the shape of AGS cells became flattened, and this morphological change was accompanied by decrease in Myc expression and an increase in the giant cell population. Lithium chloride treatment, which inhibits GSK3beta activity, reversed these serum starvation effects, which suggests an inverse relationship between Wnt signaling and giant cell formation. Furthermore, the down-regulation of Wnt signaling caused by the over-expression of ICAT, E cadherin, and Axin enhanced giant cell formation. Therefore, down-regulation of Wnt signaling may be related to giant cell formation, which is considered to be a survival mechanism against induced cell death. PMID- 15878527 TI - Distinct IkappaB kinase regulation in adult T cell leukemia and HTLV-I transformed cells. AB - We have recently shown constitutive IkappaB kinase (IKK) activation and aberrant p52 expression in adult T cell leukemia (ATL) cells that do not express human T cell leukemia virus type I (HTLV-I) Tax, but the mechanism of IKK activation in these cells has remained unknown. Here, we demonstrate distinct regulation of IKK activity in ATL and HTLV-I-transformed T cells in response to protein synthesis inhibition or arsenite treatment. Protein synthesis inhibition for 4 h by cycloheximide (CHX) barely affects IKK activity in Tax-positive HTLV-I transformed cells, while it diminishes IKK activity in Tax-negative ATL cells. Treatment of ATL cells with a proteasome inhibitor MG132 prior to protein synthesis inhibition reverses the inhibitory effect of CHX, and MG132 alone greatly enhances IKK activity. In addition, treatment of HTLV-I-transformed cells with arsenite for 1 h results in down-regulation of IKK activity without affecting Tax expression, while 8 h of arsenite treatment does not impair IKK activity in ATL cells. These results indicate that a labile protein sensitive to proteasome-dependent degradation governs IKK activation in ATL cells, and suggest a molecular mechanism of IKK activation in ATL cells distinct from that in HTLV-I transformed T cells. PMID- 15878529 TI - Registries versus trials for the evaluation of the endovascular treatment of abdominal aortic aneurysms. PMID- 15878530 TI - Results from the prospective registry of endovascular treatment of abdominal aortic aneurysms (RETA): mid term results to five years. AB - OBJECTIVES: To assess the mid-term outcomes up to 5 years following endovascular repair of abdominal aortic aneurysms (EVAR), following its initial introduction into practice in the UK. DESIGN: A prospective voluntary Registry of Endovascular Treatment of Aneurysms (RETA) collected demographic and risk factor data, short term (30 day) outcomes and follow up outcomes up to 5 years from the 41 centres that initially undertook EVAR in the UK. RESULTS: Short term outcomes (30 days): 90.4% of aneurysms were successfully excluded, 6.1% had persistent endoleaks and 5.8% of patients had died. Follow up was obtained from 30 days up to 5 years (mean 3.1 years). Returns rates for requested follow up data were 87% at 1 year and 77, 65, 52 and 51% at 2, 3, 4 and 5 years, respectively. Ninety percent of deaths at follow up were unrelated to the stent-graft or aneurysm. Persistent proximal type I endoleak was associated with significant mortality both from attempted open repair or from rupture if untreated. Other endoleaks were more benign. Complications related to the aneurysm or device occurred at an average rate of 15% per annum. The most common complications were secondary endoleaks or graft migration. Endovascular treatment was preferred if treatment was necessary for graft complications. The cumulative freedom from secondary procedure (Kaplan Meier) were 87, 77, 70, 65 and 62% at 1, 2, 3, 4 and 5 years of follow up, respectively. CONCLUSIONS: Registry data provides useful information to guide the design of more formal trials. Collecting follow up from voluntarily submitted data is difficult. The registry data remains well ahead of the trial data, but indicate that long term follow up is required in these trials, because of the high rate of complications seen at follow up. PMID- 15878531 TI - Quality of data reported on abdominal aortic aneurysm repair--a comparison between a national vascular and a national administrative registry. AB - OBJECTIVE: To study consistency of data and completeness of reporting in a national vascular registry, NorKar, and a national administrative registry, The Norwegian patient register (NPR). DESIGN: Comparative registry-based national study supplemented with a comprehensive control of patients registered in one major hospital. MATERIAL: All patients registered with a procedure-code for treatment of AAA in NorKar or NPR during 2001 or 2002, were included. METHOD: We compared the reporting of procedure-codes, diagnosis-codes and in-hospital deaths after treatment for abdominal aortic aneurysm (AAA) in the two registries to evaluate completeness. Consistency between procedure-codes and diagnoses were evaluated within both registries. Completeness of reporting to one NorKar Local Registry was investigated in more detail in one of the hospitals. RESULTS: Compared with the NPR numbers, NorKar contained 69% of the patients treated for AAA in Norway, while completeness for NorKar member hospitals was 84%. The detailed investigation in one of the hospitals showed a completeness of 91% and a false inclusion of 5.3% of all cases treated for AAA. The consistency between procedure-codes and diagnosis-codes was 93% in both registries. We found evidence of substantial underreporting of in-hospital deaths to NorKar in several hospitals. Overall reporting of early deaths to NorKar relative to completeness of reported cases was estimated to 72%. CONCLUSION: There is an underreporting of patients with AAA to NorKar according to the NPR numbers and a need for better control of procedure-diagnosis consistency in both registries. There seems to be a substantial underreporting of early deaths to NorKar. Introduction of unique patient-identifiable data could improve the quality of both registries by making matching of data possible. PMID- 15878532 TI - Single center experience with a new commercially available thoracic endovascular graft. AB - PURPOSE: To evaluate the intra-operative performance and clinical outcome of a new commercially available stent-graft for the treatment of thoracic aortic diseases. METHODS AND PATIENTS: From January 2003 to October 2004, 45 consecutive patients received endovascular treatment with the Zenith TX1 device for diseases of the thoracic aorta at a single center in northern Italy. Indications included disease of the descending thoracic aorta in 26 cases, of the aortic arch in 17 cases and of the thoraco-abdominal aorta in two cases. We treated 38 atherosclerotic aneurysms, two post-traumatic aortic ruptures, two penetrating ulcers, two chronic dissections and one case was treated for aortic bleeding after voluntary acid ingestion for attempted suicide. General anesthesia was used in 20 cases. Combined or hybrid endovascular and open surgical repair was performed in 11 patients. Mean follow-up was 7 months (range 1-22 months). RESULTS: Technical success was obtained in 44 patients (98%). One primary type I endoleak occurred (2%). ICU was used in 12 cases with a mean stay of 1 day. The mean hospital stay was 6 days (range 4-13 days). There were no hospital deaths or strokes but one transient paraplegia (2%). A type II endoleak was observed in one case and resolved spontaneously 1 month later. No aneurysm enlargement, endograft migration or structural failures were observed during follow-up. Two late unrelated-deaths were observed. CONCLUSIONS: This stent-graft does not fulfill all the characteristics of the ideal graft, however, it proved to be safe and allowed satisfactory short term results in this group of patients treated at a single center. PMID- 15878533 TI - Robot-assisted laparoscopic aortobifemoral bypass for aortoiliac occlusive disease: early clinical experience. AB - BACKGROUND: Robotic technology may facilitate laparoscopic aortic reconstruction. We present our early clinical experience with laparoscopic aortobifemoral bypass, aided by two different robotic surgical systems. METHODS: Between February 2002 and April 2004, we performed eight robot-assisted laparoscopic aorto-bifemoral bypasses for aortoiliac occlusive disease. All patients were male; median age was 55 years (range: 36-64). Dissection was performed laparoscopically and the robotic system was used to construct the aortic anastomosis. RESULTS: A robot assisted anastomosis was successfully performed in seven patients. Median operative time was 405 min (range: 260-589), with a median clamp-time of 111 min (range: 85-205). Median blood loss was 900 ml (range: 200-5800). Median anastomosis time was 74 min (range 40-110). In two patients conversion was necessary, one due to bleeding of an earlier clipped lumbar artery after completion of the anastomosis, the other because of difficulties with the laparoscopic exposure of the aorta. On post-operative day 3 one patient died unexpectedly as a result of a massive myocardial infarction. Median hospital stay was 7.5 days (range: 3-57). CONCLUSION: Our initial experience with robotic assisted laparoscopic surgery (RALS) shows it is a feasible technique for aortoiliac bypass surgery. However, laparoscopic aortoiliac surgery demands considerable experience and operative times need to be reduced before this technique can be widely implemented. PMID- 15878534 TI - Complications related to intra-aortic balloon pump in cardiac surgery: a decade later. AB - BACKGROUND: Our centre in 1995 reported 26% of vascular complications in cardiac surgical patients treated with intra-aortic balloon pump (IABP). However, during the last decade there have been improvements in IABP technology and insertion techniques. We aimed to evaluate the impact of these changes on the incidence of IABP-related complications in cardiac surgery. METHODS: Demographics, indications, technique and complication rate in 186 consecutive patients treated with IABP from January 1994 to December 1998 (Group I) were compared with 323 consecutive patients treated with IABP from January 1999 to December 2003 (Group II) at our regional cardiothoracic centre. Data was variably expressed as mean with or without range and either standard deviation or range. Statistical significance was accepted at P<0.05. RESULTS: There were 121 (65%) and 194 (60%) males in Group I and II, respectively. The mean age was 66+/-12.1 (17-88) years and the mean duration of IAPB use was 43.5h (range 3-144 h). Overall complication rate was 10% in Group I and 2% in Group II whereas vascular complications accounted for 3% in Group-I and 1% in Group-II. Logistic regression analysis demonstrated cardiogenic shock being strongly correlated to in-hospital mortality (OR 4.68; P=0.004) followed by older age (OR 3.12; P=0.034) and ejection fraction <35% (OR 1.78; P=0.03). CONCLUSION: The study demonstrated a significant decrease in the IABP-related complications even though complexity of cases referred for surgery has increased. Henceforth, the risk of 1% vascular complications should play little influence on decision-making regarding the use of IABP. PMID- 15878535 TI - Low-energy blunt abdominal aortic trauma in an underweighted man. PMID- 15878536 TI - The role of carotid endarterectomy in the endovascular era. AB - OBJECTIVE: Carotid artery angioplasty and stenting (CAS) has been proposed as an alternative to surgery for patients with high-grade symptomatic carotid disease. The purpose of this study was to determine the proportion of patients that were suitable for each type of intervention and to analyse the reasons that precluded stenting. MATERIALS AND METHODS: This was a prospective observational study. All patients considered for intervention for carotid artery disease during an 18 month period were analysed. The management decision was recorded, as were the reasons for unsuitability for stenting. RESULTS: Two hundred and sixty-eight patients' data were analysed, 224 had complete records. Forty-seven patients did not require intervention and received best medical treatment alone. One hundred and seventy-seven patients required intervention, 113 were suitable for stenting and 64 were not. In 51 patients stenting was preferred. Sixty-two patients were suitable for either stent or surgery. Sixty-four patients were unsuitable for stenting. Carotid tortuosity and proximal disease accounted for 70% of this group. CONCLUSIONS: Current enthusiasm for carotid stenting might well be supported by the results of ongoing randomised-controlled clinical trials. However, this study highlights a significant proportion (64/177; 36%) of our patients is presently unsuitable for stenting. The common technical difficulties and limitations of stenting encountered in our unit are related predominantly to carotid anatomy. PMID- 15878537 TI - In-stent restenosis after carotid angioplasty and stenting: a challenge for the vascular surgeon. AB - PURPOSE: This study aims to review the incidence of in-stent restenosis (ISR), the factors which determine restenosis, and to evaluate the use of various endovascular techniques for the management of ISR following carotid artery stenting (CAS). METHODS: Four hundred and seven patients (334 men, mean age 63 years, range 46-86, median 65 years) were treated with CAS between December 2000 and March 2004. Three hundred and seventy-two (89%) patients had at least one ultrasound evaluation performed 6 months after procedure (range 6-40). Recurrent stenosis >80% detected with duplex ultrasound scans were further evaluated by angiography and treated with repeat endovascular procedure. RESULTS: CAS was performed successfully in all cases with a Carotid WallStent (Boston Scientific) using a cerebral protection device (filter). Perioperative complications included four (0.9%) minor and two (0.4%) major strokes these latter two patients died at 5 and 12 days after the operation. No other deaths occurred. A total of 15 carotid arteries (3.6%) in 14 patients had ISR. All ISR were treated with a repeat endovascular procedure: three balloon angioplasty alone, eight angioplasty and secondary stenting, four angioplasty with cutting balloon. Postsurgical restenosis was confirmed to be the only predictive factor for the development of in-stent restenosis (OR 14.5, 95% CI 2.3-113.4, p=0.005). Endovascular treatment of ISR achieved technical success without periprocedurale complications and the absence of significant restenosis over a median follow up time of 12.4 months (range 3.5-30.7). CONCLUSION: Our experience with a large cohort of CAS showed an encouragingly low incidence of ISR (3.6%) and successful treatment by repeat endovascular intervention. We recommend attempting all endovascular possibilities before performing stent removal. PMID- 15878538 TI - The retrojugular approach to carotid endarterectomy--a safer technique? AB - The conventional technique of carotid endarterectomy involves approaching the carotid sheath, anterior and medial to the internal jugular vein with division of the facial vein. Mobilisation of the ansa cervicalis and identification of the hypoglossal nerve is usually required. We describe our results of retrojugular approach in a consecutive nonrandomised cohort of 50 carotid endarterectomy patients. PMID- 15878539 TI - Collateral artery aneurysm: a unique presentation of thoracic outlet syndrome. AB - Aneurysms of collateral arteries are unusual. A case of transverse cervical artery aneurysm as the sole presentation of vascular thoracic outlet syndrome is presented and the relevant literature reviewed. PMID- 15878540 TI - Excimer laser assisted angioplasty for critical limb ischemia: results of the LACI Belgium Study. AB - BACKGROUND: The purpose of this study was to assess the safety and efficacy of translating into national practice methodology for infrainguinal excimer laser assisted angioplasty, for the treatment of critical limb ischemia in poor surgical bypass candidates. METHODS: A prospective five centre Belgian registry enrolled 48 patients, who presented with 51 chronic critically ischemic limbs (Rutherford category 4, 5 or 6) and were poor candidates for bypass surgery. Treatment included crossing the occlusion or stenosis by conventional guidewire followed by excimer laser angioplasty with, or without, adjunctive balloon angioplasty or stenting. A step-by-step technique was used in cases where the guidewire could not pass the occluded site. The primary endpoint was limb salvage, at 6 months, of the treated limb. RESULTS: Initial treatment was successful in all 51 limbs. By 6 months there had been six deaths, six minor and four major amputations and further intervention was required in four patients. Among survivors, limb salvage rate at 6 month was 38/42 (90.5%), with freedom from critical limb ischemia in 86%. CONCLUSIONS: This Belgian study of excimer laser assisted angioplasty, in high-risk patients who were poor candidates for surgical re-vascularisation, had a low incidence of surgical re-interventions and limb salvage rate in excess of 90%. PMID- 15878541 TI - Peripheral angioplasty as the first-choice revascularization procedure in diabetic patients with critical limb ischemia: prospective study of 993 consecutive patients hospitalized and followed between 1999 and 2003. AB - OBJECTIVE: To evaluate the effectiveness of peripheral angioplasty (PTA) as the first-choice revascularisation procedure in diabetic patients with critical limb ischemia (CLI). DESIGN: Prospective study. METHODS: PTA was employed as first choice revascularisation in a consecutive series of diabetic patients hospitalized for CLI between January 1999 and December 2003. RESULTS: PTA was successful performed in 993 patients. Seventeen (1.7%) major amputations were carried out. One death and 33 non-fatal complications were observed. Mean follow up was 26+/-15 months. Clinical restenosis was observed in 87 patients. The 5 years primary patency was 88%, 95% CI 86-91%. During follow-up 119 (12.0%) patients died at a rate of 6.7% per year. CONCLUSIONS: PTA as the first choice revascularisation procedure is feasible, safe and effective for limb salvage in a high percentage of diabetic patients. Clinical restenosis was an infrequent event and PTA could successfully be repeated in most cases. PMID- 15878542 TI - Vascular PET prostheses surface modification with cyclodextrin coating: development of a new drug delivery system. AB - PURPOSE: Cyclodextrins (CDs) are torus shaped cyclic oligosaccharides with a hydrophobic internal cavity and a hydrophilic external surface. We performed and analysed an antibiotic binding on Dacron (polyethyleneterephtalate, PET) vascular grafts, previously coated with CDs based polymers. METHODS: The CDs coating process was based on the pad-dry-cure method patented in our laboratory. The Dacron prostheses were immersed into a solution containing a polycarboxylic acid, a cyclodextrin and a catalyst, and placed into a thermofixation oven before impregnation with an antibiotic solution (Vancomycin). Biocompatibility tests were performed with L132 human epithelial cells. The antibiotic release in an aqueous medium was assessed by batch type experiments using UV spectroscopy. RESULTS: Viability tests confirmed that the CDs polymers coating the Dacron fibers were not toxic towards L132 cell. Cell proliferation was similar on coated and uncoated grafts. A linear release of Vancomycin was observed over 50 days. CONCLUSION: Our results demonstrate the feasibility of coating CDs onto vascular Dacron grafts. Biological tests show no toxicity of the different cyclodextrins coated. A linear release of antibiotics was depicted over 50 days, demonstrating that cyclodextrin grafting was an efficient drug delivery system. PMID- 15878543 TI - Contemporary series of morbidity and mortality after lower limb amputation. AB - OBJECTIVES: This study was performed in order to assess morbidity and mortality associated with major lower extremity amputation according to an extensive complication registration system used in our hospital. METHODS: All consecutive patients who underwent lower limb major extremity amputation were included from January 1996 until December 2002. Complications were prospectively registered according to our standard complication registration system. RESULTS: In 97 patients 122 amputations were performed including 45 above (AKA) and 77 below (BKA) knee amputations. The conversion rate from below to above knee amputation was 14%. In 65 patients 107 complications occurred (67%). The incidence of wound infection was 10% in the BKA group and 2% in the AKA group. The most frequently reported complications were pressure sores (8%) or originating from the urinary tract (13%). The hospital mortality for BKA was 9% and for AKA 18%. Long-term survival was 62% at 1 year, 50% at 2 years and 29% at 5 years. CONCLUSIONS: An extensive registration system provides us with a detailed insight into the incidence, consequence and cause of complications. Major lower extremity amputations are still associated with considerable morbidity and mortality. PMID- 15878544 TI - Long-term treatment of deep venous thrombosis with a low molecular weight heparin (tinzaparin): a prospective randomized trial. AB - OBJECTIVES: Evaluation of the effectiveness and safety of the low molecular weight heparin (LMWH) tinzaparin versus unfractionated heparin (UFH) followed by acenocoumarol in proximal deep venous thrombosis (DVT). DESIGN: Prospective, randomized clinical trial. MATERIAL AND METHODS: Consecutive patients (n=108) with acute leg DVT, confirmed by duplex, were randomized to either tinzaparin alone or UFH and acenocoumarol for 6 months. Patients were evaluated ultrasonographically at entry, 1, 3, 6 and 12 months. Thrombus regression, reflux distribution and the incidence of complications were studied. A cost-analysis, comparing the two treatments, was performed. RESULTS: The overall incidence of major events (mortality, DVT recurrence, pulmonary embolism, major bleeding, heparin-induced thrombocytopenia) was significantly different (p=0.035) in favor of tinzaparin (7 versus 17 events). The ultrasonographic clot volume score (an index of recanalization) decreased significantly in both treatment groups. However, tinzaparin produced significantly more extended overall recanalization from 3 months onwards (p<0.02). Thrombus regression was equivalent or in favor of tinzaparin in the different DVT subgroups and venous segments, but the statistical significance varied. Reflux showed non-significant differences overall or in subgroups. A cost-analysis resulted in favor of LMWH. CONCLUSIONS: A fixed daily dose of tinzaparin for 6 months was at least as effective and safe as UFH and acenocoumarol. Regarding major events and recanalization, there was a significant benefit in favor of tinzaparin. Long-term DVT treatment with tinzaparin could represent an alternative to conventional treatment. PMID- 15878545 TI - Knot in the cava--an unusual complication of swan-ganz catheters. AB - Complications associated with the use of the Swan-Ganz catheters including coiling and knotting of the catheter in the central venous system or in the chambers of the heart, often with disastrous consequences. We report a case of knotting of a Swan-Ganz catheter in the superior vena cava which necessitated surgical removal via the right internal jugular vein. PMID- 15878546 TI - Regarding 'Life-style modification in peripheral arterial disease'. PMID- 15878548 TI - Seasonal variation in occurrence of vascular diseases. PMID- 15878550 TI - Elke Deuerling: winner of the 2005 FEBS Letters Award for Young Scientists [interview by Tine Walma]. PMID- 15878551 TI - Recombinant human alphaA-crystallin can protect the enzymatic activity of CpUDG against thermal inactivation. AB - alpha-Crystallin is one of the major protein components in mammalian lens fiber cells. It is composed of alphaA and alphaB subunits that have structural homology to the family of mammalian small heat shock proteins. Horwitz firstly characterized native alpha-crystallin as a molecular chaperone in vitro based on its ability to prevent heat-induced aggregation of lens proteins and enzymes. Andley et al. cloned and expressed human alphaA-crystallin in Escherichia coli and confirmed its chaperone activity by suppression of thermal aggregation and singlet oxygen-induced opacification. Although alphaA-crystallin acts as a chaperone protein, there is no report showing on its ability to protect enzymes against thermal inactivation. Here, we present data showing that alphaA crystallin can prevent thermal inactivation of CpUDG that catalyzes uracil removal from DNAs. PMID- 15878552 TI - The flavin reductase ActVB from Streptomyces coelicolor: characterization of the electron transferase activity of the flavoprotein form. AB - The flavin reductase ActVB is involved in the last step of actinorhodin biosynthesis in Streptomyces coelicolor. Although ActVB can be isolated with some FMN bound, this form was not involved in the flavin reductase activity. By studying the ferric reductase activity of ActVB, we show that its FMN-bound form exhibits a proper enzymatic activity of reduction of iron complexes by NADH. This shows that ActVB active site exhibits a dual property with regard to the FMN. It can use it as a substrate that goes in and off the active site or as a cofactor to provide an electron transferase activity to the polypeptide. PMID- 15878553 TI - Prediction of siRNA functionality using generalized string kernel and support vector machine. AB - Small interfering RNAs (siRNAs) are becoming widely used for sequence-specific gene silencing in mammalian cells, but designing an effective siRNA is still a challenging task. In this study, we developed an algorithm for predicting siRNA functionality by using generalized string kernel (GSK) combined with support vector machine (SVM). With GSK, siRNA sequences were represented as vectors in a multi-dimensional feature space according to the numbers of subsequences in each siRNA, and subsequently classified with SVM into effective or ineffective siRNAs. We applied this algorithm to published siRNAs, and could classify effective and ineffective siRNAs with 90.6%, 86.2% accuracy, respectively. PMID- 15878554 TI - Identification of the residue responsible for catalysing regeneration of activity in the inactive glutamate dehydrogenase mutant D165N. AB - Previously a mutant of clostridial glutamate dehydrogenase with the catalytic Asp 165 replaced by Asn was shown to regain activity through spontaneous, specific deamidation of this residue. A double mutant D165N/K125A has now been constructed, in which the catalytic Lys is also replaced. This was successfully over-expressed and according to several criteria appears to be correctly folded. The double mutant was incubated for 35 days under conditions where D165N reactivates. LC-MS analysis of tryptic digests of timed samples showed no significant deamidation. This confirms that the reactivation of D165N is a consequence of the catalytic chemistry of the enzyme's active site. PMID- 15878555 TI - Heart rate turbulence and left ventricular ejection fraction in Chagas disease. AB - AIMS: Chagas disease patients often present premature ventricular complexes (PVCs), depression of left ventricular ejection fraction (LVEF) and autonomic dysfunction, which is generally evaluated by heart rate variability (HRV) analysis. As frequent PVCs may complicate HRV computation, we measured heart rate turbulence (HRT) and evaluated the correlation between ejection fraction and HRT or HRV in Chagas disease. METHODS: We studied 30 patients (47+/-11 years, 20 men) with Chagas cardiomyopathy and left ventricular dilatation who underwent clinical evaluation, ejection fraction (EF: 45+/-14%) determination and 24-h Holter monitoring (median PVC=1781). In all patients, the standard deviation of normal RR intervals (SDNN), the square root of the mean square differences of successive RR intervals (RMSSD) and values of turbulence onset (TO) and turbulence slope (TS) were calculated. RESULTS: HRT indices were independent of mean RR interval and presented high correlation with EF: TO (-0.11+/-0.01%, r=-0.60, P<0.001) and TS (5.8+/-3.7 ms/RR-interval, r=0.73, P<0.001). Of HRV parameters, only SDNN, corrected for mean RR interval, showed a weak but not significant correlation with EF (r=0.41). The comparison of HRT/EF and HRV/EF correlation coefficients, indicated the presence of a significant difference (P=0.017). CONCLUSIONS: HRT indices appear to correlate better with EF than SDNN in Chagas disease. Thus, an analysis based on heart rate transient adaptation seems to perform better than HRV in detecting the autonomic alterations that parallel left ventricular dysfunction in Chagas disease patients. The high number of PVCs observed in these patients further support the use of HRT methodology. PMID- 15878556 TI - Improvement in sympatho-vagal imbalance and heart rate variability in patients with mitral stenosis after percutaneous balloon commissurotomy. AB - Elevated sympathetic nerve activity in patients with mitral stenosis (MS) may be an index of the severity of the disease. Percutaneous mitral balloon commissurotomy (PMBC) is now a standard treatment for many patients with symptomatic MS. We aimed to show the effects of PMBC on autonomic nervous system activity in the patients with MS by heart rate variability (HRV) analysis. Fifty four consecutive patients with mitral stenosis and sinus rhythm who underwent percutaneous mitral commissurotomy were enrolled. Apart from significant haemodynamic improvements, mean heart rate (HR), LF day, LF night, LF/HF day and night significantly decreased and SDNN, RMSSD, PNN50, HF day and night significantly increased in the early period after PMBC and these changes were preserved for up to one month. SDNN was positively correlated with left ventricle ejection fraction (LVEF) but negatively correlated with mean valve area (MVA), left atrial (LA) diameter and pressure, right atrial (RA) pressure; LF/HF day ratio was positively correlated with LA diameter and pressure, mean transmitral gradient and negatively correlated with LVEF; LF/HF night ratio was positively correlated with LA pressure and mean transmitral gradient. The increase in SDNN was correlated with the change in LA and RA pressure. The decrease in LF/HF ratio after PMBC was significantly correlated with the changes in the mean transmitral gradient, LA pressure and RA pressure. As a result, the heart rate variability and autonomic nervous system function in patients with mitral stenosis are correlated with the atrial pressures and left ventricular function. These parameters significantly change in the early period after PMBC and are preserved at one month. The improvement in the heart rate variability and sympatho-vagal balance are significantly affected by the early changes in atrial pressures after PMBC. PMID- 15878557 TI - The relationship between personality, socio-economic factors, acute life stress and the development, spontaneous conversion and recurrences of acute lone atrial fibrillation. AB - AIMS: The present study was designed to establish the relationship between personality factors, socio-economic factors and acute life stress with development, spontaneous cardioversion and recurrences of acute lone atrial fibrillation. METHODS: The study group consisted of 116 patients with lone atrial fibrillation cardioverted within 48h of the onset of arrhythmia; they underwent a series of cognitive tests to evaluate acute psychological stress and personality type. The socio-economic status and other covariates (alcohol consumption, smoking, and body mass index) were investigated. A control group, age- and sex matched, was selected and compared. In the logistic regression analysis, the presence of spontaneous conversion to sinus rhythm was used as the dependent variable. Independent variables were indicator variables representing categories of stress, Type A behaviour pattern, coffee consumption and body mass index. Variables considered for logistic analysis were only those with independent prognostic value. RESULTS: Type A behaviour pattern was found in 23 (20%) patients with atrial fibrillation and in 11 (9%) controls (P<0.001). The mean score among patients with atrial fibrillation was 8+/-2.7, while in control subjects it was 5.5+/-2. The mean acute life stress score among patients with atrial fibrillation was 56+/-33, while in controls it was 34+/-27 (P<0.01). Spontaneous conversion of atrial fibrillation to sinus rhythm was observed in 72 patients (63%). In univariate analysis alcohol consumption, income, education and smoking habits did not affect spontaneous conversion. High coffee consumption (OR 0.3 95% CI 0.11-0.49; P<0.008) and high body mass index were associated with a significantly greater risk of atrial fibrillation (OR 1.5 95% CI 1.2-1.7). CONCLUSIONS: Type A behaviour pattern and acute life stress affect the development and spontaneous conversion of atrial fibrillation. Patients with acute stress showed the highest probability of spontaneous conversion followed by patients with Type A behaviour. Other socio-economic factors affect spontaneous conversion and recurrences of lone atrial fibrillation to a lesser extent. PMID- 15878558 TI - Atrial fibrillation triggered by postinfarction ventricular premature beats in a patient with Wolff-Parkinson-White syndrome. AB - The mechanism by which atrial fibrillation is initiated in patients with accessory pathways is not fully understood. Retrograde conduction of ventricular premature beats to the atrium, causing the arrhythmia, is a very rare cause. We report a patient with Wolff-Parkinson-White syndrome (WPW), without previous tachycardias, who presented multiple episodes of paroxysmal atrial fibrillation after having a myocardial infarction. During the electrophysiological (EP) study the patient presented two spontaneous episodes of atrial fibrillation initiated by ventricular premature beats conducted to the atria through the accessory pathway. After successful catheter ablation of the accessory pathway the patient did not present arrhythmia recurrences. PMID- 15878559 TI - Transient complete atrioventricular block associated with herb intake. AB - We report a case of transient complete atrioventricular block in a 38-year-old man, after intake of a mixture of herbs, intended to aid cigarette smoking cessation. Since all other causes of conduction disturbances were excluded, a side-effect of the herbal remedy was identified as the most likely diagnosis. Given that most patients are unaware of the potential risks of the intake of various herbs, we would urge that their usage be regulated. PMID- 15878560 TI - Doxorubicin-induced second degree and complete atrioventricular block. AB - Doxorubicin is one of the most effective chemotherapeutic agents used in the treatment of malignancies. Cardiotoxicity is the most important dose-limiting toxicity of doxorubicin. Although cardiomyopathy is the most well known side effect of doxorubicin, it usually occurs many years after the treatment and relates to cumulative doxorubicin dosage. Another form of doxorubicin cardiotoxicity is arrhythmia which may occur at any time and after any dosage. However, doxorubicin-induced arrhythmia is rarely a life-threatening side effect. In this report, we present a case in which there were doxorubicin-induced life threatening arrhythmias. PMID- 15878561 TI - Orthostatic changes in AV node conduction causing dizziness during paroxysmal atrial tachycardia. AB - Dizziness in the upright posture is a common reason to seek medical advice and is often dismissed as benign orthostatic hypotension. We present an unusual case of dizziness in the upright posture due to possible changes in atrioventricular nodal conduction during atrial tachycardia. PMID- 15878562 TI - Rate-responsive pacing regulated by cardiac haemodynamics. AB - AIMS: Trans-valvular impedance (TVI) recording has been proposed for the assessment of cardiac haemodynamics, assuming an inverse relationship between TVI and ventricular volume. We checked whether the TVI sensor can drive the rate responsive function of a cardiac pacemaker following changes in the inotropic regulation of the heart. METHODS: An external DDD-R pacemaker (Ext Sophos by Medico, Padova, Italy) equipped with the TVI detecting system was tested in 30 patients on the implantation of conventional pacing leads for dual-chamber pacing. Pacing rate regulation was based on the relationship between the stroke volume and the end-diastolic volume, inferred from TVI data. After sensor calibration in basal conditions, beta-adrenergic stimulation was induced by i.v. administration of 2 microg/ml/min isoprenaline (isoproterenol) (IPN). The actual cardiac rate, the TVI waveform, the end-diastolic and systolic TVI in each cardiac cycle and the TVI-indicated rate were stored in memory as a function of time and down-loaded at the end of the session. RESULTS: All patients with intrinsic atrial activity (28/30) showed a positive chronotropic response to IPN, coupled with a significant increase in end-diastolic TVI and a four-times larger increase in end-systolic TVI. The TVI inotropic index mirrored the sinus rate time-course, with a linear correlation between the two parameters (r(2)>0.7 in 25/28 cases). As a result, the TVI-indicated rate closely reproduced the sinus rate. CONCLUSIONS: The study confirms the reliability of the haemodynamic information derived from TVI and supports its application in the regulation of rate-responsive pacing. PMID- 15878563 TI - Automatic analysis of pacemaker diagnostic data in the identification of atrial tachyarrhythmias in patients with no prior history of them. AB - AIMS: Modern pacemakers provide a large amount of diagnostic data. Given the limited time available during a pacemaker follow-up visit essential information may be overlooked. This registry was conducted to assess the utility of an expert system that analyses the diagnostic data collected by an implanted pacing device and notifies and advises the physician about suspected technical issues and arrhythmias that need further attention. METHODS: Patients with various standard indications for pacing were included in this registry and received single or dual chamber pacemakers. Data were collected and analysed by the expert system during at least two subsequent follow-up visits. The evaluation of this system focused on data obtained from patients with a dual chamber pacing device without prior history of atrial arrhythmias. RESULTS: A total number of 239 patients without prior history of atrial tachyarrhythmias were included in this analysis. Atrial tachyarrhythmias were detected in 73 (31%) of these patients. The highest incidence of newly detected arrhythmias occurred in the group of patients with high-degree AV block and VDD pacemakers. Furthermore, newly detected arrhythmias predominantly occurred in the period shortly after implantation. Device programming suggestions by the expert system were adopted in 30% of the cases. Following detection of atrial tachyarrhythmias by the expert system, pharmacological management was adjusted at 71% of the first follow-up visits and at 27% of later follow-up visits. CONCLUSION: Results of this registry show that this expert system provides a valuable tool for the detection of atrial tachyarrhythmias during pacemaker follow-up visits. PMID- 15878564 TI - Comparison of immediate and delayed automatic antitachycardia pacing for the termination of atrial tachyarrhythmias. AB - AIMS: Automatic atrial antitachycardia pacing (ATP) can terminate atrial tachyarrhythmias (ATs) in patients with an implanted device. We investigated if the programmable delay between AT onset and ATP influences therapy efficacy. METHODS: Patients with intermittent ATs and an implanted DDDRP pacemaker were randomized to receive ATP either immediately or 30min after AT detection. After four months stored data were interrogated, AT-related symptoms were assessed, and patients crossed over to the alternative treatment arm for another four months. Stored atrial electrograms were analyzed for degree of AT organization and ATP success. RESULTS: In 22 patients (64% male; 72+/-7 years), ATP success rates were higher during immediate than during delayed ATP (device classification: 59+/-7% vs. 22+/-5%, P<0.01; manual analysis: 36+/-6% vs. 12+/-5%, P<0.01). Higher efficacy of immediate ATP was associated with a larger proportion of organized (Type I) AT prior to therapy (71% vs. 44% during delayed ATP). No difference was found in total AT numbers and duration, AT burden or related symptoms. CONCLUSIONS: The programmable delay between arrhythmia onset and therapy delivery significantly influences the success-rate of ATP. However, a higher efficacy of immediate compared with delayed ATP does not translate into a reduction of AT burden or related symptoms. PMID- 15878565 TI - Pacing with capture verification in candidates for resynchronisation therapy: a feasibility study. AB - BACKGROUND: Devices for cardiac resynchronisation therapy (CRT) deliver energy into 3 output channels. Such a burden can significantly reduce device longevity. Autocapture has been shown to improve pacemaker longevity and safety of right ventricular pacing in clinical studies. The aim of this study was to investigate the application of Autocapture during biventricular pacing (BIV) to decrease the energy cost of CRT. METHODS: During implantation of BIV devices, an acute study was performed to test the hypothesis that the evoked response (ER) elicited by each delivered stimulus is correctly detected and measured either on the right ventricular (RV) channel during BIV pacing with the left ventricular (LV) channel pacing first, or in the LV channel with the RV channel pacing first. A reliable measurement of ER is the critical requirement for the correct performance of Autocapture. RESULTS: ER amplitude in the right ventricle during BIV pacing was not significantly decreased compared with RV pacing in the VVI mode (16.36+/-5.27 mV vs 17.09+/-6.12 mV). ER amplitude in the left ventricle during BIV pacing was not significantly decreased compared with LV pacing in the VVI mode (12.4+/-8.95 mV vs 12.25+/-8.97 mV). Three patients in atrial fibrillation had a DDDR pacemaker with the LV lead connected to the atrial port, and received BIV pacing with Autocapture turned on in the RV channel. Autocapture performance in the long term, as assessed by the trend of RV threshold over 20+/-8 months, showed that LV depolarisation was never sensed as an ER on the RV channel. CONCLUSIONS: Our observations support the feasibility and safety of capture verification during BIV pacing on the ventricular channel paced secondly, which could increase the longevity of CRT devices, and decrease the costs of this new therapy for heart failure patients. PMID- 15878566 TI - The incidence of implantable cardioverter defibrillator indications in patients admitted to all coronary care units in a single district. AB - AIMS: Implantable cardioverter defibrillators (ICDs) have been shown to reduce all-cause mortality in groups of patients at high risk of ventricular arrhythmias. The true incidence of ICD indications is unknown but we hypothesize that it far exceeds the number actually implanted. METHODS: In a one month observational audit, we reviewed the clinical records of all 336 patients admitted to coronary care units serving a district with a population of 471,000, to determine the additional screening tests required in myocardial infarction (MI) survivors and the number of additional ICDs which would be implanted for the primary and secondary prevention indications recommended in UK National Institute for Clinical Excellence (NICE) guidance. A further analysis was performed to determine the effect of extending the primary prevention indications to include the selection criteria used in the second multicentre automatic defibrillator trial, MADIT II. RESULTS: Using NICE criteria, we found the incidence of ICD indications to be 98.4/10(6)/year. The addition of patients fulfilling MADIT II selection criteria for primary prevention would have increased this to 453/10(6)/year. CONCLUSION: We conclude that the implementation of national guidance on the use of ICDs for arrhythmias will require the systematic screening of MI survivors, and would identify an incidence of ICD indications at least three times that anticipated by NICE, and eight times as many as were actually implanted in the UK in 2000. If the primary prevention indications were widened to include MADIT II selection criteria, the total would be nearer to 12 times that anticipated, and 47 times the number actually implanted. PMID- 15878567 TI - The usage and diagnostic yield of the implantable loop-recorder in detection of the mechanism of syncope and in guiding effective antiarrhythmic therapy in older people. AB - OBJECTIVES: To evaluate the usage and diagnostic yield of the implantable loop recorder (ILR) in detection of the mechanism of syncope and in guiding therapy in patients aged >/=65 years and comparing them with those <65 years. DESIGN: This was a two-hospital, observational, prospective study in consecutive patients with unexplained syncope who underwent an ILR implantation. Between November 1997 and December 2002, a total of 2052 patients with syncope were evaluated (local population of 590,000 inhabitants). The diagnosis remained unexplained in 371 (18%). Of these, 103 patients (5% of the total, 28% of those with unexplained syncope) received an ILR. RESULTS: There were 70 (76%) patients aged >/=65 years and 25 (24%) <65 years. ILR implantation was 110 and 9 per million inhabitants per year, respectively. During a mean follow-up of 14+/-10 months, syncope was recorded in 52 patients. Compared with younger patients those older had a 2.7 higher syncope recurrence rate (56% vs 32%, P=0.03); arrhythmias were 3.1 times more likely to be responsible for syncope (44% vs 20%, P=0.03). More patients >/=65 years finally received ILR-guided therapy (42% vs 20%, P=0.04). Among the 29 patients (25 of those >/=65 years) who received specific antiarrhythmic therapy, only one (3%), had recurrence of syncope during the subsequent follow-up of 40+/-18 months. CONCLUSIONS: In patients referred for investigation of unexplained syncope, the older subjects are more likely to have an indication for ILR implantation than those younger, ILR has a higher diagnostic value, an arrhythmia is more likely to be detected and successfully treated. PMID- 15878568 TI - Report and abstracts for Cardiostim 2005. Pacing and Haemodynamics: Art of Timing. Budapest, Hungary, April 6-8, 2005. PMID- 15878569 TI - Heterocyclic inhibitors of AChE acylation and peripheral sites. AB - Notwithstanding the criticism to the so called " cholinergic hypothesis", the therapeutic strategies for the treatment of Alzheimer's disease (AD) have been mainly centered on the restoration of cholinergic functionality and, until the last year, the only drugs licensed for the management of AD were the acetycholinesterase (AChE) inhibitors. Target enzyme AChE consists of a narrow gorge with two separate ligand binding sites: an acylation site at the bottom of the gorge containing the catalytic triad and a peripheral site located at the gorge rim, which encompasses binding sites for allosteric ligands. The aim of this short review is to update the knowledge on heterocyclic AChE inhibitors able to interact with the two sites of enzymes, structurally related to the well known inhibitors physostigmine, rivastigmine and propidium. The therapeutic potential of the dual site inhibithors in inhibiting amyloid-beta aggregatrion and deposition is also briefly summarised. PMID- 15878570 TI - Neuroendocrine correlates of behavioral polymorphism in white-throated sparrows. AB - Interspecific differences in the neuropeptide systems of the lateral septum (LS) often parallel differences in social behavior. In rodents, some closely related species that differ in aggressive behavior also differ according to the level of vasopressin (VP) innervation of the LS. In songbirds, the neuropeptides vasotocin (VT) and vasoactive intestinal peptide (VIP) affect aggression when administered directly to the LS. Here, we tested whether the density of VT or VIP innervation of the LS reflects patterns of intraspecific behavioral polymorphism in male and female white-throated sparrows (Zonotrichia albicollis), in which the "white stripe" (WS) morph behaves more aggressively than the "tan-stripe" (TS) morph. We found that the WS birds had more VT-immunoreactivity (IR) than the TS birds in the ventrolateral subdivision of the caudal LS (LSc.vl) and in the medial portion of the bed nucleus of the stria terminalis (BSTm). In addition, the TS birds had more densely stained VIP-IR in the LSc.vl than the WS birds. Males had more VT-IR than females in the LSc.vl and BSTm, and more VIP-IR in the LSc.vl. We also report sex and morph differences in VIP-IR in the basal hypothalamus, where VIP is synthesized and released into the portal vasculature. Males had nearly twice as many VIP-immunoreactive (ir) neurons in the infundibular nucleus than did females, and birds of the WS morph had more densely stained VIP-IR in the median eminence than TS birds. Our results support the hypothesis that differences in these neuropeptide systems underlie inter- and intraspecific differences in social behavior across vertebrates. PMID- 15878571 TI - Cognitive abilities, androgen levels, and body mass index in 5-year-old children. AB - This study explores the potential relationship between a series of cognitive abilities and testosterone, dehydroepiandrosterone (DHEA), androstenedione, and body mass index (BMI) measurements in 5-year-old children. 60 boys and 69 girls were administered a test (K-BIT) which provided measurements of fluid intelligence (Matrices subtest), crystallized intelligence (Vocabulary subtest), and IQ composite (the combination of the two subtests); a sub-sample of 48 boys and 61 girls was also subjected to diverse tests related to theory of mind (affective labeling, appearance-reality distinction, display rules, and false belief). Testosterone, DHEA, and androstenedione levels were measured using an enzyme immunoassay technique in saliva samples. An analysis of variance failed to reveal any significant differences between boys and girls in any of the cognitive abilities assessed. The correlation analysis revealed a positive relationship between fluid intelligence and testosterone levels in boys, a negative relationship between crystallized intelligence and androstenedione levels in girls, and between affective labeling and androstenedione levels in boys. A multiple regression analysis indicated that androstenedione and BMI were the best predictors for some of the cognitive abilities assessed. PMID- 15878572 TI - Tactile contact is required for early estrogen treatment to alter the sexual partner preference of female zebra finches. AB - Sex steroid actions during early development appear to play a role in the development of sex differences in sexual partner preference (SPP, preference for males vs. females) in several species of mammals and in the socially monogamous pair bonding zebra finch (Taeniopygia guttata). Female finches treated with estrogen as nestlings exhibit varying degrees of masculinized SPP as adults, but only if they have been housed in all-female groups during the juvenile and young adult period, suggesting that the estrogen effect may involve social experience and possibly sexual imprinting. Because tactile contact is important for consolidation of imprinted preferences in this species, it was predicted that early estrogen treatment would alter preferences of females only if they were allowed to have tactile contact with other females. Subjects were injected with estradiol benzoate or with oil (normal controls) daily for the first 2 weeks post hatching. At age 45 days, they were housed in a mixed sex aviary (normal controls), in an all-female aviary allowing tactile contact (group EB-TC), or in an all-female aviary with no tactile contact (group EB-NTC). At 100+ days, birds were given two-choice SPP tests followed by aviary tests of SPP. EB-TC females did not show the sex-typical preference for male stimuli, and differed significantly from the controls on several measures. EB-NTC females preferred males and never differed significantly from controls. These results show that tactile contact after age 45 days is essential for an EB effect on SPP, supporting the hypothesis that hormones and sexual imprinting together contribute to SPP. PMID- 15878573 TI - Experimental manipulation of yolk testosterone affects digit length ratios in the ring-necked pheasant (Phasianus colchicus). AB - In humans, most of the mammals and one bird species studied so far, the relative length of individual digits is sexually dimorphic. Most studies of humans have been concerned with the ratio between second (2D) and fourth digits (4D), whereas some studies of humans and other mammals have also investigated other digit ratios. Inter- and intra-sexual variation in 2D:4D may depend on differential exposure to androgens during embryonic life, and the genetic mechanisms linking 2D:4D to androgens may be mediated by Hox genes. Because Hox genes are conserved in vertebrates, similar patterns of variation in digit ratios might be expected across vertebrate classes. The observation of correlations between digit ratios and physiological, psychological and performance traits in humans has generated interest in exploring the possibility that digit ratios are a marker of embryonic exposure to androgens, which have diverse consequences on several phenotypic traits. However, the hypothesis that digit ratios depend on androgen effects during development has never been tested experimentally. In this study, we increased testosterone concentration in ring-necked pheasant eggs and measured length ratios between the second, third and fourth digits of both feet in fully grown offspring. Females from testosterone-injected eggs had larger 2D:3D in the left foot, whereas this was not the case in males. The other digit ratios were unaffected by hormone treatment in both sexes. However, digit ratios showed no sexual dimorphism among controls. Thus, present results are consistent with the hypothesis that variation in testosterone levels during development affects digit ratios. PMID- 15878574 TI - Lactation reduces prolactin levels in reproductively experienced female rats. AB - Long-term alterations in prolactin (PRL) secretion following reproductive experience have been demonstrated in both women and female rats. In the rat, these changes include decreased PRL secretion in response to a dopamine antagonist challenge following ovariectomy, decreased post-coital diurnal and nocturnal prolactin surges in multigravid versus primigravid females, as well as decreased suckling-induced prolactin release in multiparous versus primiparous females. To date, there have been no studies examining PRL secretion following reproductive experience in cycling female rats. Studies in women, however, have demonstrated a reduction in basal PRL secretion during the menstrual cycle. The purpose of the present work was to determine whether similar changes occur in the rat during the estrous cycle and to what extent lactation is involved in these effects. In addition to examining PRL, potential parity-induced changes in estradiol secretion were also studied. The findings revealed a significant decrease in PRL levels during the afternoon of proestrus, which was only observed in primiparous females that had lactated. Significant differences in estradiol secretion were not detected following reproductive experience. Thus, a reduction in the PRL surge on the afternoon of proestrus is a consequence of reproductive experience that requires both pregnancy and lactation. PMID- 15878575 TI - Conservation and behavioral neuroendocrinology. AB - The total number of threatened species of vertebrates is likely to be more than 10,000, with approximately one quarter of the world's mammal species, one eighth of the birds and one third of the amphibians threatened with extinction. The rate of loss of animal species and hence of biodiversity is increasing and may become even greater as ecosystems become affected by climate change due to global warming. Behavioral neuroendocrinology, which considers interactions between behavior and neuroendocrine function in animals from all vertebrate taxa, can contribute to animal conservation. Research with laboratory animals can address questions in basic biology relevant to conservation and develop methods for use with threatened animals. Field work with free-living animals considers the basic biology of new species and the use of endocrine tools to assess the susceptibility of species to threats. Non-invasive measurements of hormone concentrations, especially fecal steroids, are extensively used to assess reproductive function and the stress status of animals in captive breeding programs and in the wild. Biodiversity and natural selection both depend on individual variation, and conservation programs often work with animals on an individual basis. The consideration of data from individuals is essential in conservation endocrinology. Direct contributions to conservation programs are challenging as study situations are determined by practical conservation concerns. Indirect contributions such as the provision of scientific input to conservation plans and participation in public education programs offer significant benefits for conservation programs. Directly and indirectly, there are many opportunities for behavioral neuroendocrinologists to contribute to conservation. PMID- 15878576 TI - Anabolic androgenic steroid affects social aggression and fear-related behaviors in male pair-housed rats. AB - This study examines the effect of chronic administration of the anabolic androgenic steroid nandrolone decanoate (ND) on dominant and subordinate male rats in a pair-housed condition. Pair-housed rats were assessed for dominance status based on their behavior and alterations in body weights. Throughout the study the rats were allowed limited social interactions on a daily basis. At all other times, a Plexiglas divider kept the rats separated, allowing olfactory and visual contact between the cage mates while preventing significant physical contact. One week into the study all subjects were subcutaneously implanted with a pellet that continuously infused either ND (15 mg/kg/day) or placebo for 21 days. Following the pellet implant, behavioral tests including reassessment for dominance status, and a conditioned fear test were conducted over a period of approximately 2 months to investigate possible long-term changes. The main finding is that during the allowed social interactions, the dominant ND pretreated rats spent more time on highly aggressive behaviors than the dominant placebo-treated rats. In addition, the probability for highly aggressive behaviors was maintained for the ND-treated rats throughout the study, whereas it was decreased for the placebo-treated rats. The ND-treated subordinate rats showed less fear in a potential threatening situation compared to placebo-treated controls. These findings support the relatively long-term behavioral changes that have been seen in humans after abuse of ND and other anabolic androgenic steroid compounds. PMID- 15878577 TI - The hormonal control of begging and early aggressive behavior: experiments in black-headed gull chicks. AB - The hormonal control of begging and sibling competition is largely unknown, but recent evidence suggests a role for steroid hormones. We tested the influence of the aromatizable androgen testosterone (T), the non-aromatizable androgen 5alpha dihydrotestosterone (DHT), and 17beta-estradiol (E) on both begging behavior and aggressive behavior in black-headed gull chicks (Larus ridibundus). Chicks of this species have a conspicuous begging display, while their frequently performed early aggressive behavior is facilitated by testosterone and important for territorial defense. Hormone treatment was applied by implants between days 6 and 16 after hatching. Behavior was tested by means of standard stimulus tests. The results were validated in a second experiment under semi-natural conditions. Begging was suppressed by T and DHT and not affected by E. Aggressive Pecking was strongly facilitated by T. The erect threat posture, characteristic for older chicks, was facilitated by T, DHT, and E and the nest-oriented threat display, typical for young chicks, only by T and DHT. Growth was suppressed in the T group. The results indicate that androgen production, needed for territorial defense, has costs in terms of a suppression of begging and growth. It is discussed to what extent older chicks may avoid these costs by converting testosterone to estrogen and why pre-natal and post-natal exposure to androgens differ in their effect on begging behavior. PMID- 15878578 TI - Social competition and plasma testosterone profile in domesticated canaries: an experimental test of the challenge hypothesis. AB - The challenge hypothesis predicts that plasma testosterone (T) concentration is high when male-male competitions are high and decreases when males are engaged in paternal care. In monogamous species, T concentration increases at the beginning of the breeding period and decreases after egg laying. According to the challenge hypothesis, increasing competition should also lead to T increase. The aim of our study was to test this hypothesis. In a first experiment, we measured the T profile of domesticated canaries housed with their mate in separated cages without competition. In a second one, we created a competition by housing male and female domestic canaries together (in an aviary) and emphasized this competition by limiting food access. We also studied social status effect. Our results showed no effect of social status in both sexes and no differences in female's T concentration. Concerning males, we obtained a clear monogamous T profile from the ones housed in a low competition situation and a polygamous profile from the others housed in high competition situation. Thus, our results support the hypothesis of the plasticity of the mechanisms controlling T concentration according to environmental conditions. PMID- 15878579 TI - Simultaneous treatment with an aromatase inhibitor and an anti-androgen decreases the likelihood of dawn song in free-living male great tits, Parus major. AB - Gonadal steroids, most importantly testosterone (T), are considered to be a major factor in the expression of adult song behavior in temperate-zone songbirds. The action of T within specific brain regions involved in the regulation of song may occur either directly, or through its androgenic or estrogenic metabolites. In the present study, we tested steroid-dependence of great tit dawn song by blocking both known pathways of T action by simultaneous implantation of flutamide, an anti-androgen, and ATD, an aromatase inhibitor. By our knowledge, this is the first study investigating the effects of androgen inhibitors on dawn song in free-living birds. Male great tits were implanted during their mate's egg laying stage, being the time of maximal male song activity at dawn. Treatment with ATD and flutamide significantly increased plasma T levels, probably because feedback mechanisms on T secretion were inhibited. The treatment decreased the likelihood of showing dawn song, which is in line with the hypothesis that sex steroids are involved in the endocrine control of song behavior. In males that did show dawn song, we found no evidence for a treatment effect on song quality. Although the implants were present for the larger part of the breeding season, males were able to maintain control of a territory and mate and to complete their brood cycle as successful as control males. PMID- 15878580 TI - The effect of tau genotype on clinical features in FTDP-17. AB - The clinical phenotype of frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17) varies. This variability is seen not only between kindreds with different mutations but also in families sharing the same mutation. Inheritance of tau haplotype (H1) and genotype (H1/H1) has been established as a risk factor for some neurodegenerative disorders with parkinsonism. We assessed the effect of tau polymorphism on the clinical features of FTDP-17 in 61 cases from 30 separately ascertained families with four different tau mutations, including P301L, +16, N279K, and P301S. There were no significant differences of age at symptomatic onset and disease duration between H1/H1 and H1/H2 genotypes. The comparison between tau genotype and type of initial clinical sign showed an association between the H1/H1 genotype and parkinsonian phenotype and between the H1/H2 genotype and frontotemporal dementia phenotype (OR=11.7; 95% confidence interval, 1.4-98.7; P=0.008). Our results suggest that tau genotype does not influence the disease course. However, it may predispose to a specific clinical sign in the early stage of FTDP-17. PMID- 15878581 TI - Visual event-related potentials under different interstimulus intervals in Parkinson's disease: relation to motor disability, WAIS-R, and regional cerebral blood flow. AB - We have introduced S1-S2 paradigm (task S) as well as oddball paradigm (task O) visual event-related potentials (ERPs) under different interstimulus intervals (ISIs) in Parkinson's disease (PD). ERP measurements were correlated with motor disability, WAIS-R, and regional cerebral blood flow (rCBF). The 'group' influence was characterized by longer latency for P300, N200, and reaction time and decreased P300 amplitude in PD. Both P300 latency and reaction time during task O showed significantly longer latency in longer ISI condition. Our results revealed 'ISI' influence on ERPs during task S and significant correlation between ERPs and rCBF in task S. PMID- 15878582 TI - Quality of life in patients with Parkinson's disease. AB - We evaluated the quality of life (QoL) in 278 Indian patients with Parkinson's disease (PD) using PDQL questionnaire, with measures various aspects of health status in PD patients including parkinsonian symptoms, systemic symptoms, social and emotional functions. Apart from demographic and treatment details parkinsonian disability and stage was assessed by Hoehn and Yahr stage, Schwab and England scale and UPDRS scores. We conclude that female gender, presence of depression, low degree of independence, higher levodopa dose (>400 mg/day) and higher UPDRS activity of daily living score have the most detrimental impact on QoL in patients with Parkinson's disease. Depression was the most significant factor impairing the QoL and it needs to be treated. PMID- 15878583 TI - Brain perfusion differences between Parkinson's disease and multiple system atrophy with predominant parkinsonian features. AB - BACKGROUND: The patterns of regional cerebral blood flow in Parkinson's disease and multiple system atrophy remain inconsistent. OBJECTIVES: To compare brain perfusion images of 123I-IMP SPECT between Parkinson's disease, multiple system atrophy with predominant parkinsonian features (MSA-P) and controls. METHODS: Eighty-two patients with Parkinson's disease, 10 patients with MSA-P and 14 controls were studied. We performed 3D-SSP and volume of interest analysis using 123I-IMP scintigraphy. RESULTS: Occipital perfusion of MSA-P increased compared to that of Parkinson's disease and perfusion in the cerebellum and primary sensorimotor cortex of Parkinson's disease increased compared to that of MSA-P. Perfusion in the putamen of MSA-P decreased compared to that of Parkinson's disease. CONCLUSION: Our study demonstrated perfusion differences in 123I-IMP SPECT between the two diseases. PMID- 15878584 TI - Prevalence of essential tremor in Singapore: a study on three races in an Asian country. AB - To investigate the prevalence of Essential Tremor (ET) in Singapore and compare the rates between Singaporean Chinese, Malays, and Indians, a community-based survey among a disproportionate random sample of 15,000 individuals (9000 Chinese, 3000 Malays, 3000 Indians) aged 50 years and above was conducted. In phase 1, trained interviewers conducted a door-to-door survey using a screening questionnaire for Parkinson's disease. In phase 2, medical specialists examined participants who screened positive to evaluate for the presence of postural or kinetic tremor of the upper limbs, or head tremor. Participants with suspected ET had their diagnosis confirmed in phase 3 by a movement disorders specialist and fellow based on the latest core diagnostic criteria. Forty participants with classic ET were identified. The prevalence rate (PR) of ET was 2.37 per 1000 (95% CI: 1.65-3.32), age-adjusted to UICC world standard population. The PR was significantly higher in males (p=0.01) and increased significantly with age (p<0.001). Indians (PR=4.94 per 1000, 95% CI: 2.63-9.04) were 1.8 times more likely to have ET than Chinese (PR=2.77 per 1000, 95% CI: 1.78-4.17) (p=0.08). No Malays with ET were identified. The data suggest that the prevalence of ET increases with age, is higher in males and may be higher amongst Indians. PMID- 15878585 TI - Behind the facial twitch: depressive symptoms in hemifacial spasm. AB - BACKGROUND: Depression impairs psychosocial and occupational functioning and contributes to significant morbidity and mortality. Hemifacial spasm (HFS) causes social embarrassment and visual and verbal disability. OBJECTIVE: We examined; (1) the prevalence and predictive factors of depressive symptoms (Becks Depression Inventory (BDI) and clinical assessment) in HFS and (2) the sensitivity and specificity of BDI as a screening and diagnostic tool in HFS. METHODS: A large cohort of HFS patients in a movement disorders clinic was clinically evaluated and the BDI self-administered by patients. Univariate analysis and multivariate logistic regression were undertaken to investigate the effect of age, gender, body-mass index, duration and severity of HFS on the outcome of BDI score. ROC (receiver operating characteristics) analysis was utilized to evaluate the sensitivity and specificity and discriminative property of the scale. RESULTS: There were 90 HFS patients with a mean age of 54.4+11.1 (35-79) years, comprising of 58.9% women and with a mean severity HFS score of 2.9+0.8 (range 1-4). The mean BDI score was higher in depressed HFS than in non depressed HFS (19.7+6.7 vs 4.2+4.9, p<0.0001). Female gender and a younger age were risk factors (p=0.07). In the multivariate analysis, the severity of HFS was an independent predictor of BDI scores (p<0.0001). The AUC was 97.1% suggesting excellent discriminative property of BDI. For cut-off score of 12/13, the sensitivity was 93.3%, specificity 94.7%, Positive Predictive Value 77.8% and Negative Predictive Value 98.6%. CONCLUSIONS: The prevalence of depressive disorder in HFS was 16.7%, with younger women at greater risk. The severity of HFS was positively correlated with the severity of depressive symptoms. The BDI can be a complimentary screening and/or diagnostic instrument for depressive disorder in HFS. Early diagnosis of at-risk patients will prevent unnecessary morbidity and mortality. PMID- 15878586 TI - Weight and body mass index in Parkinson's disease patients after deep brain stimulation surgery. AB - A retrospective chart review characterizing changes in 17 male and 10 female Parkinson's disease (PD) patients undergoing deep brain stimulation (DBS) surgery indicated that 6 mo before surgery, patients lost a mean of 5.1 lbs, whereas in the 6 mo after surgery, subjects gained a mean of 10.1 lbs; 22% gained more than 14 lbs. In 10 patients followed an additional 6 mo, weight gain continued. This weight gain may be associated with decreased energy expenditure due to subsidence of chronic tremor. The magnitude of gain underscores the need for proactive management of body weight in PD patients undergoing DBS. PMID- 15878587 TI - Reduced plasma homocysteine levels in levodopa/entacapone treated Parkinson patients. AB - Hyperhomocysteinemia is not only a major risk factor for atherothrombotic disease, but is also strongly associated with an increased risk of dementia and cognitive impairment, both of which are common in the course of Parkinson's disease (PD). Previous work has found that levodopa increases plasma homocysteine concentrations. Animal studies have indicated that the catechol-O methyltransferase (COMT) inhibitors can prevent levodopa-induced elevation of homocysteine concentrations by reducing the O-methylation of levodopa. The objective of our study was to assess the impact of entacapone, a COMT inhibitor, on plasma levels of homocysteine, serum folate, and vitamin B12 in levodopa treated PD patients. Nineteen PD patients receiving only levodopa and 21 PD patients on a combination of levodopa and entacapone participated in the cross sectional study. The control group consisted of 17 subjects on dopamine agonists. The mean plasma homocysteine concentration in the subjects on only levodopa was higher than that in the subjects on a combination of levodopa and entacapone (P=0.001) or in the control group (P=0.0001). Concentrations of serum vitamin B12 and serum folate were on average normal in all groups, but levodopa-treated subjects (with or without entacapone therapy) were more prone to have hypovitaminosis B12 (45%) than controls on dopamine agonists (6%). We suggest that the COMT inhibition may play a promising role in successfully controlling levodopa-induced hyperhomocysteinemia and in reducing the risk of pathologies probably linked to it. These preliminary findings and postulated hypotheses must now be confirmed in prospective studies. PMID- 15878588 TI - Unilateral subthalamic nucleus deep brain stimulation contralateral to thalamic stimulation in Parkinson disease. AB - The effects of unilateral subthalamic nucleus (STN) stimulation contralateral to thalamic stimulation in Parkinson disease (PD) have not been previously reported. We are reporting a patient who developed left arm tremor in 1994, at age 62, as her first PD symptom. She underwent right thalamic DBS surgery in 1999 that resulted in complete resolution of left arm tremor. Her PD symptoms progressed and she developed severe motor fluctuations and disabling dyskinesias. In 2003, she underwent left STN electrode implantation. Left STN stimulation improved contralateral motor scores in the medication OFF state, and allowed for reduced medication doses and less dyskinesia. However, there was no significant improvement in activities of daily living (ADL), motor scores in the medication ON state, gait, or postural stability. PMID- 15878589 TI - Deep brain stimulation in a patient on immunosuppressive therapy after renal transplant. AB - We performed thalamic deep brain stimulation (DBS) surgery to treat severe essential tremor in a 36 year-old woman who had undergone cadaveric renal transplant four years earlier. She was receiving chronic immunosuppressive therapy. Post-operative healing was normal and there have been no infections of the DBS hardware. There were no peri-operative complications and no rejection of the transplanted kidney. She remains on the same systemic immunosuppressive agents as pre-operatively: prednisone, cyclosporine, and mycophenolate mofetil (CellCept). DBS surgery may be safely performed in carefully selected patients on systemic immunosuppression after renal transplant. PMID- 15878590 TI - Long-term treatment of sleep breathing disorder in a patient with Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder. An association with sleep breathing disorder has not previously been established in the literature. We report the case of a 45-year-old woman with HD, presenting with snoring and observed apnea. Polysomnography showed obstructive sleep apnea with an apnea-hypopnea index (AHI) of 6.6 per hour including events up to 57 s long during rapid eye movement (REM) sleep. A trial of continuous positive airway pressure (CPAP) resulted in significant improvement in sleep structure, nocturnal respiration, daytime alertness, and subjective memory. Snoring, apneas, and respiratory arousals were abolished on CPAP. HD patients may suffer from sleep breathing disorder, and in spite of potential chorea, CPAP is feasible and may significantly improve their quality of life. PMID- 15878591 TI - Administration of selegiline is not associated with visual hallucinations in advanced Parkinson's disease patients. PMID- 15878593 TI - Numerical analysis of maximal bat performance in baseball. AB - Metal baseball bats have been experimentally demonstrated to produce higher ball exit velocity (BEV) than wooden bats. In the United States, all bats are subject to BEV tests using hitting machines that rotate the bat in a horizontal plane. In this paper, a model of bat-ball impact was developed based on 3-D translational and rotational kinematics of a swing performed by high-level players. The model was designed to simulate the maximal performance of specific models of a wooden bat and a metal bat when swung by a player, and included material properties and kinematics specific to each bat. Impact dynamics were quantified using the finite element method (ANSYS/LSDYNA, version 6.1). Maximum BEV from both a metal (61.5 m/s) and a wooden (50.9 m/s) bat exceeded the 43.1 m/s threshold by which bats are certified as appropriate for commercial sale. The lower BEV from the wooden bat was attributed to a lower pre-impact bat linear velocity, and a more oblique impact that resulted in a greater proportion of BEV being lost to lateral and vertical motion. The results demonstrate the importance of factoring bat linear velocity and spatial orientation into tests of maximal bat performance, and have implications for the design of metal baseball bats. PMID- 15878594 TI - Glycation increases human annulus fibrosus stiffness in both experimental measurements and theoretical predictions. AB - One of the primary age-related changes to collagenous tissues is the increased concentration of advanced glycation endproducts (AGEs). Although AGEs have been shown to increase the mechanical stiffness of many tissues, their influence on the mechanical properties of the annulus fibrosus has not been measured experimentally. In previous theoretical work, we hypothesized that the mechanical influence of AGEs on the annulus could be represented in an additive strain energy function with a separate crosslinking term, but the material coefficients associated with this term were not correlated with AGE concentration. In the current study, we measured the tensile stress-strain response of the human annulus in the axial direction both before and after glycation with methylglyoxal. Using nonlinear regression, the strain energy function was simultaneously applied to these new data and to data from a wide range of experimental protocols reported in the literature to determine values for the material coefficients appearing in the constitutive equation. Nonenzymatic collagen crosslinking induced a statistically significant change in annular material properties. Furthermore, the concentration of AGEs correlated positively with the material coefficients found in the terms of the strain energy function that we associate with collagen crosslinking. These data suggest that AGEs contribute to age-related disc stiffening as well as validate the hypothesis that biochemical constituents can be related mathematically to tissue behavior. In the future, this structurally guided constitutive relationship may provide further insight into the structure-function relationships of the annulus fibrosus. PMID- 15878595 TI - The crystal structure of Plasmodium falciparum glutamate dehydrogenase, a putative target for novel antimalarial drugs. AB - Plasmodium falciparum is the main causative agent of tropical malaria, the most severe parasitic disease in the world. Growing resistance of Plasmodia towards available drugs is an increasing problem in countries where malaria is endemic. As Plasmodia are sensitive to oxidative stress, augmenting this in the parasite represents a promising principle for the development of novel antimalarial drugs. The NADP-dependent glutamate dehydrogenase (GDH) of P.falciparum is largely responsible for the production of NADPH in the parasite, which in turn serves as electron source for the antioxidative enzymes glutathione reductase and thioredoxin reductase. As GDH does not occur in the host erythrocyte, GDH is a particularly attractive target for drug therapy. The three-dimensional structure of P.falciparum GDH in the unligated state has been determined by X-ray crystallography to a resolution of 2.7A. Compared to the mammalian enzymes, two amino acid residues are exchanged in the putative active site of the parasite GDH. The most obvious differences between parasite and human GDH are the subunit interfaces of the hexameric proteins. In the parasite protein, several salt bridges mediate contacts between the subunits whereas in the human enzyme these interactions are mainly of hydrophobic nature. Furthermore, P.falciparum GDH possesses a unique N-terminal extension that does not occur in any other GDH sequence so far studied. These findings might be exploited for the design of peptidomimetics capable of disrupting the oligomeric organisation of the parasite enzyme. PMID- 15878596 TI - Physical coupling between ryanodine receptor-calcium release channels. AB - Ryanodine receptor-calcium release channels play a pivotal role in the calcium signaling that mediates muscle excitation-contraction coupling. Their membrane organization into regular patterns, functional gating studies and theoretical analysis of receptor clustering have led to models that invoke allosteric interaction between individual channel oligomers as a critical mechanism for control of calcium release. Here we show that in reconstituted "checkerboard like" lattices that mimic in situ membrane channel arrays, each oligomer is interlocked physically with four adjacent oligomers via a specific domain-domain interaction. Direct physical coupling between ryanodine receptors provides structural evidence for an inter-oligomer allosteric mechanism in channel regulation. Therefore, in addition to established cytosolic and luminal regulation of function, these observations indicate that channel-channel communication through physical coupling provides a novel mode of regulation of intracellular calcium release channels. PMID- 15878597 TI - Effect on fatality risk of changing from secondary to primary seat belt enforcement. AB - BACKGROUND: Most seat belt use laws originally passed in the United States contained language restricting enforcement to drivers already stopped for some other reason. States that have since removed this secondary enforcement restriction have reported increased seat belt use. The purpose of the present study was to estimate the effect of these law changes on driver fatality rates. METHOD: Trends in passenger vehicle driver death rates per billion miles traveled were compared for 10 states that changed from secondary to primary seat belt enforcement and 14 states that remained with secondary enforcement. RESULTS: After accounting for possible economic effects and other general time trends, the change from secondary to primary enforcement was found to reduce annual passenger vehicle driver death rates by an estimated 7% (95% confidence limits 3.0-10.9). CONCLUSION: The majority of U.S. states still have secondary enforcement laws. If these remaining secondary laws were amended, an estimated 696 deaths per year could be prevented. PMID- 15878598 TI - Steroid hormones in multiple sclerosis. AB - The possible influence of steroid hormones in multiple sclerosis (MS) has been a matter of great interest. A first illustration comes from the analyses of the influence of gender on susceptibility to MS and on MS severity. A series of arguments emerge in favour of a possible influence of steroid hormones in MS. The menstrual cycle, and even more pregnancy, may influence the clinical evolution of MS. In the PRIMS study, there was a dramatic decrease in the relapse rate during pregnancy, especially in the third trimester, with a rebound increase in the 3 months post partum. Animal studies have provided further confirmatory results. Many experiments have shown that sex steroids may have immunological effects, in preventing or treating experimental allergic encephalomyelitis. They could also have an effect on myelinating and remyelinating the peripheral and possibly the central nervous system. These clinical and experimental data have led to consider sexual steroids as potential therapeutic tools for preventing relapses in women with MS, in particular in the post-partum period. PMID- 15878599 TI - Down-regulation of the sodium channel Na(v)1.1 alpha-subunit following focal ischemic brain injury in rats: in situ hybridization and immunohistochemical analysis. AB - Change in sodium channel (NaCh) activity can play a role in reorganization, recovery, or possibly excitotoxic damage after CNS injury. Alteration of sodium channel function has been reported to occur in a variety of neuropathological states including epilepsy and brain injury. Previously we reported that out of five NaCh alpha subunit genes that were down-regulated, Na(v)1.1 exhibited the most dramatic and sustained alterations following focal cerebral ischemia in the rat. In the present study, we evaluated the acute spatial and temporal time course distribution of Na(v)1.1 mRNA (in situ hybridization) and protein (immunohistochemistry) following ischemic brain injury. Male rats were subjected to 2 h of middle cerebral artery occlusion (MCAo) followed by reperfusion and brain tissue was collected at 2, 6, 24, and 48 h post-MCAo. Analysis of brain tissue revealed a qualitative drop in both mRNA and protein levels of Na(v)1.1 throughout ischemic regions, beginning at the early stage of injury (6 h) with dramatic losses at later stages (24 and 48 h). Quantitative cell counts and optical density measurements indicated significant decreases in the percent of brain cells immunoreactive for Na(v)1.1 as well as a loss of signal in those cells positive for Na(v)1.1 in the injured cortex and striatum as compared to the contralateral hemisphere. Double labeling with NeuN and Na(v)1.1 immunoflouresence confirmed that the predominate loss of Na(v)1.1 immunoreactivity was in neurons. In conclusion, these data map the time-dependent loss of Na(v)1.1 mRNA and protein following focal ischemic brain injury in the rat out to 48 h post-injury. PMID- 15878600 TI - Endomorphin 1[psi] and endomorphin 2[psi], endomorphins analogues containing a reduced (CH2NH) amide bond between Tyr1 and Pro2, display partial agonist potency but significant antinociception. AB - Endomorphin 1 (EM1) and endomorphin 2 (EM2) are highly potent and selective mu opioid receptor agonists and have significant antinociceptive action. In the mu selective pocket of endomorphins (EMs), Pro2 residue is a spacer and directs the Tyr1 and Trp3/Phe3 side chains into the required orientation. The present work was designed to substitute the peptide bond between Tyr1 and Pro2 of EMs with a reduced (CH2NH) bond and study the agonist potency and antinociception of EM1[psi] (Tyr[psi(CH2NH)]Pro-Trp-Phe-NH2) and EM2[psi] (Tyr[psi(CH2NH)]Pro-Phe Phe-NH2). Both EM1[psi] and EM2[psi] are partial mu opioid receptor agonists showing significant loss of agonist potency in GPI assay. However, EMs[psi] exhibited potent supraspinal antinociceptive action in vivo. In the mice tail flick test, EMs[psi] (1, 5, 10 nmol/mouse, i.c.v.) produced potent and short lasting antinociception in a dose-dependent and naloxone (1 mg/kg) reversed manner. At the highest dose of 10 nmol, the effect of EM2[psi] was prolonged and more significant than that of EM2. In the rat model of formalin injection induced inflammatory pain, EMs[psi] (0.1, 1, 10 nmol/rat, i.c.v.), like EMs, exerted transient but not dose-dependent antinociception. These results suggested that in the mu-selective pocket of EMs, the rigid conformation induced by the peptide bond between Tyr1 and Pro2 is essential to regulate their agonist properties at the mu opioid receptors. However, the increased conformational flexibility induced by the reduced (CH2NH) bond made less influence on their antinociception. PMID- 15878601 TI - Phosphorus dynamics in water and sediments in urbanized and non-urbanized rivers in Southern Brazil. AB - Phosphorus flux models show that the removal tends to be a common feature in polluted estuarine systems whereas the release of P to the adjacent coastal area occurs in pristine environments. This study analyzes the distribution of P in water and sediments along six rivers in the south coast of Brazil. Three rivers located inside protected areas were considered non-polluted and used as a control of urbanization. The other three, situated within urbanized areas, were considered as having highly elevated concentrations of P. Results showed a different behavior of P in water and sediments located in urbanized and non urbanized areas. The concentrations of dissolved organic (P-org) and inorganic (P inorg) phosphorus in water, and the total phosphorus (P-tot) and polyphosphate (P poly) in sediments where higher in the urbanized rivers compared to the non urbanized ones. Both P of punctual origin and of diffuse origin contributed to the maintenance of elevated concentrations and disturbed the natural fluxes along the polluted rivers. The minimum and maximum concentrations in urbanized areas varied from 0.39 to 12.45 (microM) for P-org and 0.00 to 5.92 (microM) for P inorg in water, and in sediments from 89.90 to 808.16 (microM g(-1)) for P-tot and 0.00 to 76.51 (microM g(-1)) for P-poly. In non-urbanized areas concentrations in water varied from 0.22 to 1.20 (microM) for P-inorg and 4.43 to 5.56 (microM) for P-org, and in sediments from 45.91 to 652.26 (microM g(-1)) for P-tot and 0.00 to 8.61 (microM g(-1)) for P-poly. Using a hierarchical sampling design and a simple model of variation of P (K(d) model) it was possible to demonstrate that urbanized and non-urbanized estuaries may act as sinks or sources of P. The variation of P analysis in different points along each of the six rivers showed that release and removal areas may occur within a same river independently of its urbanization. PMID- 15878602 TI - The cerebellum's involvement in the judgment of spatial orientation: a functional magnetic resonance imaging study. AB - A functional magnetic resonance imaging (fMRI) study was conducted to integrate the clinical observations of the impaired judgment of spatial orientation of cerebellar patients with recent theoretical discoveries about the role of the cerebellum in cognitive functions. Ten normal healthy male right-handed Chinese postgraduates consented to participate in this study. The experimental task employed was a modified version of Benton's Judgment of Line Orientation Test, administered in a blocked fMRI study. The findings indicated activation of the cerebellar regions, the Hemisphere Lobules IV, VI and Crus I, while the subjects were performing the experimental task of the judgment of the orientation of lines. Furthermore, cortical regions were activated, including the bilateral precuneus (BA 7), the extrastriate regions (BA 19), and the bilateral prefrontal regions (BA 9, 10, 44, 46). The imaging data confirmed that the activity of the cerebellum is associated with judging spatial orientation. The theoretical and clinical implications of the findings are discussed. PMID- 15878603 TI - Behaviour of food restricted broilers during rearing and lay--effects of an alternative feeding method. AB - Broiler breeders are subjected to quantitative food restriction in order to control their growth, and this restriction is particularly severe during rearing. While such restriction improves some welfare problems associated with ad libitum feeding, it causes others: birds routinely show abnormal oral behaviours and have elevated plasma corticosterone concentration (PCC) and changes in white blood cell counts (WBCs). The aim of this study was to examine if feeding birds qualitatively restricted diets ad libitum during rearing could reduce signs of impaired welfare, as judged by behaviour and blood indices of stress, while also meeting commercially desired growth rates and uniformity. Furthermore, we examined what carry-over effects such a feeding method had on birds in the laying phase when all birds were fed on a conventional quantitative restriction regime. During rearing (1-20 weeks of age), pens of birds were either fed limited amounts of standard basal diets (Control, i.e. quantitative restriction), or ad libitum diets consisting of standard basal diets with gradually increasing levels of calcium propionate (CaP) and a constant level of oat hulls (OH), designated CaP+OH (i.e. qualitative restriction). Results showed that, during rearing, weights and weight uniformity were similar for the two groups. During feeding motivation tests, Control birds always consumed more food than CaP+OH birds. This suggests that Control birds were more highly motivated to feed than CaP+OH birds, although care has to be taken in interpreting these results. Treatment did not affect PCCs or WBCs, but there was a general decline in PCCs with bird age. All reported behaviours differed significantly between treatment groups during rearing, but disappeared during lay when all birds were fed a similar amount of food. Control birds spent up to 50% of time in object pecking during rearing periods, but this behaviour was virtually non-existent in birds in the qualitative feeding regime. Overall, the results indicate that broiler breeders can be successfully limited in growth rates by qualitative food restriction and this results in significant changes of behaviour that suggest improvements to bird welfare. PMID- 15878604 TI - Biotic and abiotic degradation behaviour of ethylene glycol monomethyl ether (EGME). AB - Glycol ethers are widely used in many processes in the chemical industry. Their high water solubility means they are used as solvents for different purposes (e.g. lacquers and varnishes). Since glycol ethers are known to produce toxic metabolites such as the teratogenic methoxyacetic acid during biodegradation, the biological treatment of glycol ethers can be hazardous. However, using oxidizing agents like hydrogen peroxide could be a feasible option for treating wastewater containing glycol ether. In this study, both-, biodegradation and abiotic oxidation experiments with ethylene glycol monomethyl ether (EGME) as contaminant were performed. The biodegradation experiments were conducted with a synthetic model wastewater containing 15 wt% NaCl and 5000 mgl(-1) of EGME. While experiments with the fungus Aspergillus versicolor resulted in the exhaustive biotic degradation of EGME, the toxic metabolite methoxyacetic acid (MAA) was produced as a 'dead end' product. Sodium hydroxide was added to adjust the decreasing pH caused by the production of MAA. In abiotic degradation experiments with EGME, other degradation products--organic acids and toxic aldehydes, e.g. methoxy acetaldehyde (MALD)--were detected. It must be taken into account that EGME and its biotic and abiotic degradation products are usually not analysed in routine wastewater measurements owing to their physical properties. PMID- 15878605 TI - Bioassays and selected chemical analysis of biocide-free antifouling coatings. AB - Over the years several types of biocide-free antifouling paints have entered the market. The prohibition of biocidal antifouling paints in special areas of some European countries such as Sweden, Denmark and Germany has favoured the introduction of these paints to the market. Several types of biocide-free antifouling paints were subjected to bioassays and selected chemical analysis of leachate and incorporated substances. Both non-eroding coatings (silicones, fibre coats, epoxies, polyurethane, polyvinyl) and eroding coatings (SPCs, ablative) were tested to exclude the presence of active biocides and dangerous compounds. The paints were subjected to the luminescent bacteria test and the cypris larvae settlement assay, the latter delivering information on toxicity as well as on efficacy. The following chemical analyses of selected compounds of dry-film were performed: The results of the bioassays indicated that none of the coatings analysed contained leachable biocides. Nevertheless, some products contained or leached dangerous compounds. The analyses revealed leaching of nonylphenol (up to 74.7 ng/cm2/d after 48 h) and bisphenol A (up to 2.77 ng/cm2/d after 24 h) from epoxy resins used as substitutes for antifouling paints. The heavy metal, zinc, was measured in dry paint film in quantities up to 576,000 ppm in erodable coatings, not incorporated as a biocide but to control the rate of erosion. Values for TBT in silicone elutriates were mostly below the detection limit of 0.005 mg/kg. Values for DBT ranged between <0.005 and 6.28 mg/kg, deriving from catalysts used as curing agents. Some biocide-free paints contained leachable, toxic and dangerous compounds in the dry film, some of which may act as substitutes for biocides or are incorporated as plasticizers or catalysts. Implications to environmental requirements and legislation are discussed. PMID- 15878606 TI - Photocatalytic degradation of azo dyes by nitrogen-doped TiO2 nanocatalysts. AB - This study examined the photocatalytic degradation of three azo dyes, acid orange 7 (AO7), procion red MX-5B (MX-5B) and reactive black 5 (RB5) using a new type of nitrogen-doped TiO2 nanocrystals. These newly developed doped titania nanocatalysts demonstrated high reactivity under visible light (lambda>390 nm), allowing more efficient usage of solar light. The doped titania were characterized by X-ray diffraction (XRD) and transmission electron microscopy (TEM). Experiments were conducted to compare the photocatalytic activities of nitrogen-doped TiO2 nanocatalysts and commercially available Degussa P25 powder using both UV illumination and solar light. It is shown that nitrogen-doped TiO2 after calcination had the highest photocatalytic activity among all three catalysts tested, with 95% of AO7 decolorized in 1 h under UV illumination. The doped TiO2 also exhibited substantial photocatalytic activity under direct sunlight irradiation, with 70% of the dye color removed in 1h and complete decolorization within 3 h. Degussa P25 did not cause detectable dye decolorization under identical experimental conditions using solar light. The decrease of total organic carbon (TOC) and evolution of inorganic sulfate (SO4(2 )) ions in dye solutions were measured to monitor the dye mineralization process. PMID- 15878607 TI - Long series relationships between global interannual CO2 increment and climate: evidence for stability and change in role of the tropical and boreal-temperate zones. AB - Interannual variability in global CO2 increment (averaged from the Mauna Loa and South Pole Stations) shows certain strong spatial relationships to both tropical and temperate temperatures. There is a fairly strong positive year-round correlation between tropical mean annual temperatures (leading by 4 months) and annual CO2 throughout the time series since 1960, agreeing with the generally held view that the tropics play a major role in determining inter-annual variability in CO2 increment, with a major CO2 pulse following a warm year in the tropics. This 'almost no lag' climatic response is very strong during winter and relatively stable in time. However, the correlation with tropical temperature appears to have weakened in the first years of the 1990s in correspondence of the Pinatubo eruption and the positive phase of the AO/NAO. A secondary concurrent temperature signal is linked to summer variations of north temperate belt. Northern summer temperatures in the region 30-60 degrees N-and especially in the land area corresponding to the central east USA-have become relatively more closely correlated with CO2 increment. This trend has become increasingly stronger in recent years, suggesting an increasing role for growing season processes in the northern midlatitudes in affecting global CO2 increment. Once non-lagged annual tropical temperature variations are accounted for, terrestrial ecosystems, especially the temperate-boreal biomes, also show a coherent large scale lagged response. This involves an inverse response to annual temperature of preceding years centered at around 2 years before. This lagged response is most likely linked to internal biogeochemical cycles, in particular N cycling. During the study period north boreal ecosystems show a strengthening of the lagged correlation with temperature in recent years, while the lagged correlation with areas of tropical ecosystems has weakened. Residuals from a multiple correlations based on these climatic signals are directly correlated with SO, confirming an additional important role of upwelling in interannual variability of CO2 increment. Cooler summers following the Pinatubo eruption and the possible influence of the North Atlantic Oscillation (NAO/AO) are discussed as factors responsible for the shift in the relative importance of different regions over time during the series of data. PMID- 15878608 TI - Kinetics of mass and DNA decomposition in tomato leaves. AB - This laboratory study investigated the kinetics of leaf and DNA content decomposition in two varieties of tomato (Palmiro and Admiro) after incubation in soil for 35 days. Results revealed that the decrease of dry matter in leaves in both varieties did not follow a single exponential function and was better described by a double exponential model. Composite half-decrease times were 3.4 and 2.4 days for Palmiro and Admiro respectively. The same pattern was observed for DNA mass loss, although this was closer to a single exponential model with composite half-decrease times of 1.5 and 1.4 days. Genomic analysis showed that DNA in dried leaves at room temperature (not inoculated in the soil), remains intact or presents a weak degradation, and DNA extracted from leaves inoculated in non-sterile soil showed degradation after two days. These results indicate that before release an important quantity of DNA may be degraded inside plant tissues during decomposition in soil. PMID- 15878609 TI - Factors affecting biosurfactant production by oil degrading Aeromonas spp. isolated from a tropical environment. AB - An Aeromonas spp. was isolated from tropical estuarine water. The organism grew on crude oil and produced biosurfactant that could emulsify hydrocarbons. The peak growth and biosurfactant production was on the 8th day. The organism grew on a range of hydrocarbons that include crude oil and hexadecane while no growth was recorded on some hydrocarbons that include benzene. The biosurfactant produced by the organism emulsified a range of hydrocarbons with diesel (E24=65) as the best substrate and hexane (E24=22) as the poorest. After purification, the biosurfactant was found to contain about 38% carbohydrate and an unidentified lipid. No protein was present in the purified biosurfactant. Production of biosurfactant was highest in medium with glucose and lowest in the medium with diesel+acetate. Soybean was the best nitrogen source for biosurfactant production. The activity of the biosurfactant was enhanced optimally at NaCl concentration of 5%, pH of 8.0 and temperature of 40 degrees C. The biosurfactant retained 77% of its original activity after 120 min of exposure to heat at a temperature of 100 degrees C. Biosurfactant may be produced with this organism using non-hydrocarbon substrates such as glucose and soybean that are readily available and would not require extensive purification for use in food and pharmaceutical industries. PMID- 15878610 TI - A practical approach to the degradation of polychlorinated biphenyls in transformer oil. AB - A practical and efficient disposal method for hydrodechlorination of polychlorinated biphenyls (PCBs) in transformer oil is reported. Transformer oil containing PCBs was treated by nanometric sodium hydride (nano-NaH) and transition metal catalysts. High destruction and removal efficiency (89.8%) can be attained by nano-NaH alone under mild conditions. The process exhibits apparent characteristics of a first order reaction. The reductive ability of nano NaH was enhanced by the addition of transition metal catalysts. In the presence of TiCl4, 99.9% PCBs was hydrodechlorinated. The complex reducing reagents, Ni(OAc)2+i-PrONa, show extra hydrodechlorinating activity for di-chlorinated biphenyls. PMID- 15878611 TI - Net changes in nutrient concentrations below a point source input in two streams draining catchments with contrasting land uses. AB - We examined net changes in ammonium-Nitrogen (NH(4)(+)-N), nitrate-Nitrogen (NO(3)(-)-N), dissolved inorganic nitrogen (DIN), soluble reactive phosphorus (SRP), and dissolved organic carbon (DOC) chloride-corrected ambient concentrations along a reach located below a wastewater treatment plant (WWTP) input in a non-agricultural (12 dates) and an agricultural (6 dates) stream. Based on those net changes, we estimated processing length (Snet) and mass transfer coefficient (Vf) of the cited nutrients. In the agricultural stream, results suggest that diffuse nutrient inputs from adjacent agricultural fields had a greater effect on water chemistry than the WWTP input, and probably overwhelmed the stream capacity to retain and transform nutrients. In the non agricultural stream we observed consistent longitudinal trends below the WWTP input only for NH(4)(+)-N and NO(3)(-)-N. The tight coupling between longitudinal NH(4)(+)-N decreases and NO(3)(-)-N increases in the non-agricultural stream, and lack of longitudinal trends of DIN on most dates suggest that NH(4)(+)-N from the WWTP input was being nitrified along the reach. These results suggest that WWTP inputs favor conditions to support hot spots for chemoautotrophic activity. PMID- 15878612 TI - Differential inhibitory effects of platelet glycoprotein IIb/IIIa antagonists on aggregation induced by procoagulant agonists. AB - INTRODUCTION: In addition to mediating the final common pathway of aggregation, the glycoprotein (GP) IIb/IIIa receptor participates in the activation of coagulation on the platelet surface. High-affinity conformation of GP IIb/IIIa in response to collagen-induced inside-out signalling seems to be mediated by GP VI( FcRgamma) and reinforced by release of soluble mediators. METHODS: We assessed the effects of the three currently available GP IIb/IIIa antagonists--abciximab, tirofiban and eptifibatide--on platelet aggregation induced by various procoagulant and GP VI-related agonists, i.e. collagen-related peptide (CRP), convulxin and collagen fibrils, in PPACK-anticoagulated platelet-rich plasma. RESULTS: At concentrations that equally inhibited 80% of ADP-induced maximal aggregation abciximab-inhibited GP VI-mediated platelet responses to CRP or convulxin significantly more than the low-molecular-weight antagonists (CRP: abciximab 75+/-18%, tirofiban 41+/-7% and eptifibatide 41+/-6%; convulxin: abciximab 90+/-6%, tirofiban 64+/-20%, eptifibatide 61+/-14%, p<0.01 for all). In contrast, aggregation induced by collagen was equally abolished with all antagonists under the similar conditions. During CRP- or convulxin-triggered platelet activation, inhibition of fibrin polymerisation with GPRP potentiated the antiaggregatory effects of tirofiban and eptifibatide to reach that of abciximab. GPRP as such did not affect platelet aggregation. CONCLUSIONS: GP IIb/IIIa antagonists exhibit distinct inhibition profiles in platelet aggregation, depending on fibrin polymerization and calcium. Specifically, the ability of procoagulant platelet agonists to expose pre-activated and ligand bound GP IIb/IIIa from the internal pool seems important. PMID- 15878613 TI - Distribution patterns of the anti-angiogenic protein ADAMTS-1 during rat development. AB - A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS-1) belongs to the large ADAM family of proteins. ADAMTS-1 contains a metalloproteinase domain, a disintegrin domain and three thrombospondin-like repeats but unlike ADAMs lacks a transmembrane domain. For the elucidation of the biological functions of ADAMTS-1, we raised new antibodies against ADAMTS-1. We show an accumulation of ADAMTS-1 protein at the basal lamina of rat embryonal epithelia of intestines, nasal cavity, choroid plexus, skin and in intracellular storage vesicles of epithelial cells. ADAMTS-1 protein seems to play a role in the development of the neuronal system, adipose tissue, muscle, heart, liver and adrenal glands. At the time of birth its presence is reduced in most organs. However, in the developing bone as well as in the skin, labelling increased towards late embryonal development. Immunoblots revealed a strong presence of a 62 kDa ADAMTS-1 fragment in kidneys, adrenal glands, lungs, intestines and heart. ADAMTS-1 was also present in the corresponding adult rat organs, but in more restricted distribution patterns. It was typically found in principal cells of collecting ducts, of the renal medulla, in ependymal cells of the ventricles and in some neurons. The results were confirmed by real-time PCR. The specific distribution pattern of ADAMTS-1 in a variety of organs during embryogenesis suggests a role of the molecule in tissue remodelling, vasculogenesis and angiogenesis. PMID- 15878614 TI - Light microscopical study of nitric oxide synthase I-positive neurons, including fibres in the vestibular nuclear complex of the cat. AB - Nitric oxide is a gaseous neurotransmitter that is synthesized by the enzyme nitric oxide synthase I (NOS I). At present, little is known of NOS I-positive neurons in the vestibular nuclear complex of the cat (VNCc). The aim of the present study was to examine the morphology, distribution patterns and interconnections of NOS I-positive neurons, including fibres in the VNCc. Five adult cats were used as experimental animals. All cats were anaesthetized and perfused transcardially. Brains were removed, postfixed, cut on a freezing microtome and stained in three different ways. Every third section was treated with the Nissl method, other sections were stained either histochemically for NADPH diaphorase or immunohistochemically for NOS I. The atlas of Berman (1928) was used for orientation in the morphometric study. NOS I-positive neurons and fibres were found in all parts of VNCc: medial vestibular nucleus (MVN); lateral vestibular nucleus (LVN); superior vestibular nucleus (SVN); inferior vestibular nucleus (IVN); X, Y, Z groups and Cajal's nucleus. The NOS I-positive neurons were classified according to their size (small, medium-sized, large neurons type I and type II) and their shape (oval, fusiform, triangular, pear-shaped, multipolar and irregular). In every nucleus, a specific neuronal population was observed. In SVN, a large number of interconnections between NOS I-positive neurons were identified. In MVN, chain-like rolls of small neurons were found. Tiny interconnections between MVN and mesencephalic reticular formation were present. Our data provide information on the morphology, distribution patterns and interconnections of NOS I-positive neurons in the VNCc and can be extrapolated to other mammals. PMID- 15878616 TI - An in-vitro study of the sterilization of titanium dental implants using low intensity UV-radiation. AB - OBJECTIVES: Commercial titanium dental implants are coated with nanostructured TiO2. The aim of the research reported in this paper was to assess whether the TiO2 at the surface of a dental implant is sufficiently photoactive to eradicate bacteria when illuminated with low intensity light. METHODS: The photoactivity of dental implants was established by studies of the photoenhanced decomposition of Rhodamine B. In vitro studies to establish the influence of irradiating with UV light an implant that is immersed in a solution containing Escherichia Coli were performed. RESULTS: It was demonstrated that under low UV intensity irradiation, 49 microW cm(-2), bacteria are killed at a rate of approximately 650 million per cm2 of implant per minute. SIGNIFICANCE: The results indicate that illumination of dental implants with UV light may be a suitable treatment for periimplantitis. PMID- 15878617 TI - Resin composites: strength of the bond to dentin versus surface energy parameters. AB - OBJECTIVES: The strength of the bond to dentin treated with two adhesive systems was investigated. The bond was tested with six resin composites. It was hypothesized that the surface energy parameters of composite and adhesive-treated dentin would influence the bond strength. METHODS: The investigated adhesive systems were Prompt L-Pop and OptiBond FL, which were used as recommended. The bond strengths were tested in shear. The dispersive, the acid, and the base component of the surface energy were determined from measurements of the contact angle of four test liquids placed on the composites and on the adhesive-treated dentin. Relationships were investigated by regression analysis. RESULTS: The strength of the bond to dentin varied with the resin composite and with the adhesive system. The bond strength with both adhesive systems increased with the base component of the surface energy of the resin composites. Dentin treated with OptiBond FL had higher surface energy and gave higher bond strengths than was the case with Prompt L-Pop. SIGNIFICANCE: It is advisable to use a resin composite that is compatible with the adhesive system. PMID- 15878618 TI - Fluoride release from aged resin composites containing fluoridated glass filler. AB - OBJECTIVES: The aim of this study was to evaluate the fluoride release from aged resin composites containing different types of fluoridated glass filler into both deionized distilled water and lactic acid solution. METHODS: Three resin composites, UniFil S (containing fluoro-alumino-silicate glass filler), Reactmer (containing pre-reacted glass-ionomer filler) and Beautifil (containing both types of fillers) were used. A conventional glass-ionomer cement, Ketac-Fil, was used as a control. Five disk specimens of each material were prepared and aged in water for 10 weeks. After aging, specimens were immersed in deionized distilled water for a further 6 days and then in aqueous lactic acid (pH 4.0) for 2 days. This process was repeated twice more and the specimens were subsequently immersed in water for a further 12 days. Fluoride release was measured every 2 days throughout the post-aging period. RESULTS: The amount of fluoride release for aged UniFil S and Beautifil markedly increased in acid solution compared with water storage. The difference was not so great for aged Reactmer and Ketac-Fil. UniFil S and Beautifil gave significantly greater fluoride release in water following immersion in acid solution (p<0.05, two-way ANOVA and Scheffe's test), but Reactmer and Ketac-Fil showed no such increase in fluoride release after acid immersion. SIGNIFICANCE: These results suggested that the nature of the fluoridated glass filler within a resin composite and the way in which the material interacts with an acidic environment affected the amount of fluoride released. PMID- 15878619 TI - Unpacking commitment and exploration: preliminary validation of an integrative model of late adolescent identity formation. AB - A model of identity formation comprising four structural dimensions (Commitment Making, Identification with Commitment, Exploration in Depth, and Exploration in Breadth) was developed through confirmatory factor analysis. In a sample of 565 emerging adults, this model provided a better fit than did alternative two- and three-dimensional models, thereby validating the unpacking of both exploration and commitment. Regression analyses indicated that Commitment Making was significantly related to family context in accordance with hypotheses. Identification with Commitment and both exploration dimensions were significantly related to adjustment and family context, again in accordance with hypotheses. Identification with Commitment was positively related to positive adjustment indicators and negatively to depressive symptoms, whereas Exploration in Breadth was positively related to depressive symptoms and substance use. Exploration in Depth, on the other hand, was positively related to academic adjustment and negatively to substance use. Implications and suggestions for future research are discussed. PMID- 15878620 TI - Aberrant expression of tumor suppressor genes and their association with chimeric oncogenes in pediatric acute lymphoblastic leukemia. AB - Aberrant expression of tumor suppressor genes WT 1, RB 1, p53, homozygous deletion of p16 gene and their relationship with expression of oncogenes BCR-ABL, TEL-AML 1, MLL-AF 4, E2A-PBX 1, SIL-TAL 1 were determined in bone marrow samples of children with de novo B-lineage (n=170) and T-lineage (n=25) acute lymphoblastic leukemia (ALL). In contrast to expression of chimeric oncogenes alterations in p16, WT 1, RB 1 and p53 expression were T/B-lineage-unrestricted. Significant association between expression of MLL-AF 4 and WT 1, E2A-PBX 1 and p53; SIL-TAL 1 and homozygous deletion of p16 has been demonstrated. PMID- 15878621 TI - Endotoxin- or pro-inflammatory cytokine-induced sickness behavior as an animal model of depression: focus on anhedonia. AB - In humans, exposure to endotoxins or pro-inflammatory cytokines induces a number of neuropsychological symptoms collectively referred to as 'flu-like syndrome'. The degree of overlap between flu-like syndrome and major depressive disorder is considerable and a close linkage between these has been predicted to arise due to hypersecretion of endogenous pro-inflammatory cytokines and activation of the hypothalamic pituitary adrenal axis. In animals, exposure to pro-inflammatory cytokines or endotoxins induces a 'sickness behavior' syndrome that is analogous to flu-like symptoms observed in human patients. The goal of the current paper is to review evidence implicating endotoxin- or cytokine-induced sickness behavior as an animal model of depression, with an emphasis on reduced consumption of highly palatable substances as a defining feature. PMID- 15878622 TI - Isatin-Schiff base copper(II) complexes and their influence on cellular viability. AB - Some copper(II) complexes with isatin (isa) or imine ligands derived from isatin were prepared, characterized by analytical and spectroscopic techniques, and had their biological activity toward proliferation of two different cell types verified. These complexes exhibit keto-enolic equilibria in aqueous solution, very dependent of pH, although isolated in the solid state in one defined form, and this type of equilibrium was previously verified to be crucial for their catalytic activity in the oxidation of carbohydrates, through intermediary generation of reactive oxygen species. Herein, biological studies carried out with tumor cells of different origin such as human neuroblastoma (SH-SY5Y) and promonocytic (U937) cells showed that these compounds exert different toxicity. In particular, while compounds [Cu(isaen)(H(2)O)]ClO(4).2H(2)O 2, [Cu(isahist)(H(2)O)](ClO(4))(2)4 and [Cu(isa)(2)]ClO(4)6 are not toxic for both cell lines at the concentrations used in this study, compounds [Cu(isapn)](ClO(4))(2)1, [Cu(isaepy)(2)](ClO(4))(2).2H(2)O 3 and [Cu(isami)(H(2)O)]ClO(4)5 are cytotoxic, with the compound 3 being the most effective. In these compounds, isaen, isahist, isapn, isaepy and isami stand for imine ligands prepared by condensation of ethylenediamine (en), histamine (hist), 1,3-diaminopropane (pn), 2-aminoethylpyridine (epy), and 8-aminoquinoline (ami) with isatin (isa). Cells treated with these compounds were committed to the apoptotic program as evidenced by cytofluorimetric analyses of cell cycle. Moreover, the toxicity of compound 5 was equivalent for both cell lines while the compound 1 was almost not toxic at 24h for SH-SY5Y cells where only an arrest in G1 phase was observed. Compound 3 was more efficient in inducing cell death and also in this case a striking effect on U937 cells (apoptotic cells 68% compared with 11% of SH-SY5Y) was observed. Therefore, the results indicated that their activity seems to be cell type specific. PMID- 15878623 TI - Ternary copper(II) complexes with N-carboxymethyl-l-prolinato(2-) ion and imidazole or creatinine: a comparative study of the interligand interactions influencing the molecular recognition and stability. AB - The compounds {[Cu(CMP)(Him)].H(2)O}(n) (I) and [Cu(CMP)(crea)H(2)O].3H(2)O (II) were synthesized and characterized by X-ray diffraction, thermal, spectral and magnetic methods (CMP=N-carboxymethyl-;l-prolinato(2-) ion, Him=imidazole and crea=creatinine). Appropriate structural comparison with other compounds such as {[Cu(CMP)(H(2)O)].H(2)O}(n), [Cu(crea)(2)Cl(2)] and [Cu(dipeptide)(crea)(H(2)O)(x)].nH(2)O (x=0 or 1) have been made in order to prove that crea can act as an imidazole-like ligand (because it is able to promote the same fac- to mer-CMP tridentate conformational change in copper(II) complexes) as well as to discuss the interligand interactions which control the 'Cu(CMP) complex-crea, molecular recognition processes. In contrast to that found in related ternary complexes, we have concluded that direct CMP-crea interligand interactions are missing in the Cu-CMP-crea complex due to the inappropriate correspondence between the donor and/or acceptor H-bonding properties of these ligands. CMP can only act as H-acceptor by its two terminal carboxylate group, and crea can display H-donor and H-acceptor roles by its exocyclic -NH(2) and O moieties, respectively. That promotes the reinforcement of the Cu-N(crea) bond by a bridge -N-H(crea)...O(aqua) (2.867(3)A, 176.4 degrees). PMID- 15878624 TI - Adenosine receptors and the control of endothelial cell function in inflammatory disease. AB - The nucleoside adenosine accumulates in many tissues following the onset of ischaemia and inflammation. This initiates a series of protective mechanisms in target cells upon binding and activation of a family of four G-protein-coupled cell surface adenosine receptor (AR) proteins. The magnitude and duration of adenosine's effects are dictated by the identity and expression levels of each receptor subtype on individual cell types within the hypoxic microenvironment. Given the key role of endothelial cells (ECs) in the development of inflammatory diseases, such as sepsis, rheumatoid arthritis (RA) and atherosclerosis, ARs represent attractive targets for therapeutic intervention in these conditions. In this review, we examine several critical aspects of endothelial function in vivo, assess the role of individual AR subtypes in these events and, where known, discuss the molecular mechanisms by which specific ARs exert their effects. PMID- 15878625 TI - Sox group E gene expression distinguishes different types and maturational stages of glial cells in developing chick and mouse. AB - The SoxE genes, Sox8, Sox9 and Sox10, have been shown to modulate glial specification and differentiation in the peripheral nervous system and spinal cord. However, little is known of their expression during brain development. We have therefore analyzed their expression in developing brain, focused on the cerebellum. In situ hybridization showed a striking degree of temporo-spatial variation in their expression in different glial lineages in both chick and mouse. Sox8 appeared both in oligodendrocytes and astrocytes, Sox9 was more abundant in astrocytes and Sox10 was restricted to later stages of oligodendrocyte development. PMID- 15878626 TI - Olfactory bulb mitral cell dendritic pruning abnormalities in a mouse model of the Fragile-X mental retardation syndrome: further support for FMRP's involvement in dendritic development. AB - The Fragile-X mental retardation syndrome is the leading form of inherited mental retardation. Dendritic analysis in a mouse model (FraX) found abnormal pruning in somatosensory cortex. To further characterize dendritic abnormalities and assess their occurrence in other brain regions, we examined mitral cells in FraX mice olfactory bulbs. FraX mice exhibited dendritic abnormalities consistent with somatosensory cortex, suggesting that deficient pruning is found in multiple brain regions. PMID- 15878627 TI - ADAM-17 and TIMP3 protein and mRNA expression in spinal cord white matter of rats with acute experimental autoimmune encephalomyelitis. AB - Tumour necrosis factor (TNF) is a major immunomodulatory and proinflammatory cytokine implicated in the pathogenesis of multiple sclerosis (MS) and the animal model experimental autoimmune encephalomyelitis (EAE). ADAM-17 cleaves membrane bound TNF into its soluble form. The distribution and level of ADAM-17 expression within spinal cords of Lewis rats with EAE was investigated. ADAM-17 was associated with endothelial cells in the naive and pre-disease spinal cords. In peak disease astrocytic and inflammatory cells expressed ADAM-17. Upregulation of ADAM-17 mRNA expression was coupled with a decrease in mRNA levels of its inhibitor TIMP3 suggesting a role for ADAM-17 in EAE pathogenesis. PMID- 15878628 TI - N-benzyloxycarbonyl-L-proline: an in vitro and in vivo inhibitor of prolidase. AB - Prolidase deficiency (PD) is a recessive disorder of the connective tissue caused by mutations in the prolidase, a specific peptidase, cleaving the dipeptides with a C-terminal prolyl and hydroxyprolyl residue. PD is a complex syndrome characterized mainly by intractable skin lesions, recurrent respiratory infections and mental retardation. The relation between prolidase biological functions and the disease is still largely unknown. We studied the effect of a prolidase inhibitor, N-benzyloxycarbonyl-l-proline (Cbz-Pro), in vitro on prolidase from human fibroblasts and in vivo on murine erythrocytes prolidase. A 90% inhibition was detected incubating cellular extracts at 1:1 ratio of Gly-Pro substrate: Cbz-Pro inhibitor. Pulse experiments performed incubating human fibroblasts with 6 mM Cbz-Pro revealed that the inhibitor uptake was completed in about 1 min. The Cbz-Pro uptake was saturable and pH dependent. Long-term incubation of fibroblasts with Cbz-Pro caused mitochondria depolarization and increased cellular death as reported for long-term culture of fibroblasts from PD patients. An inhibitory effect of Cbz-Pro has also been shown in vivo. Our results demonstrated that Cbz-Pro is a potent inhibitor of prolidase in cultured fibroblasts and it can be used in vivo to better characterize the prolidase enzyme and further investigate PD physiopathology. PMID- 15878629 TI - Differential effect of estrogen receptor alpha and beta agonists on the receptor for advanced glycation end product expression in human microvascular endothelial cells. AB - Estrogens are known to induce the expression of the receptor for advanced glycation end products (RAGE). In the current investigation, we examined the effect of three estrogens with different potency for specific estrogen receptors (ER) on RAGE expression in human microvascular endothelial cells (HMEC-1). Of the three estrogens tested, ethinyl estradiol (EE), an estrogen receptor alpha (ERalpha) agonist, was the strongest inducer of RAGE expression in HMEC-1. By comparison, 17-epiestriol, an estrogen receptor beta (ERbeta) agonist and 17-beta E2, an ER agonist that is almost equally potent for ERalpha and ERbeta were less effective in stimulating RAGE expression. We then determined whether the prooxidative and proinflammatory transcription factors Sp1 or NF-kappaB were downstream modulators of ER-agonists that mediate RAGE expression. The results implicated Sp1 but not NF-kappaB in estrogen-dependent RAGE expression. We further demonstrated that ERalpha but not ERbeta was responsible for the estrogen mediated Sp1 activation. In summary, the present investigation demonstrates that a direct interaction of EE-ERalpha-Sp1 plays a central role in estrogen-induced RAGE expression in HMEC-1. PMID- 15878630 TI - Testing the new ICRU 62 'Planning Organ at Risk Volume' concept for the rectum. AB - BACKGROUND AND PURPOSE: To study the impact of the new ICRU 62 'Planning organ at Risk Volume' (PRV) concept on the relationship between rectum dose-volume histogram (DVH) data and toxicity. PATIENTS AND METHODS: The acute gastro intestinal (GI) RTOG toxicity in 127 prostate cancer patients prescribed a total dose of 70 Gy with conformal irradiation to either the prostate, the prostate and seminal vesicles or the whole pelvis (initial 50 Gy only) were analysed. DVHs were derived for the rectum only and for rectum extended with six PRV margin sets (narrow/intermediate/wide; anterior/anterior and posterior). The data was analysed using permutation tests, logistic regression and effective uniform dose (EUD) calculations. RESULTS: Acute Grade 2 GI toxicity was seen in 22 of 127 cases (17%). Permutation tests showed that the difference between DVHs for patients with and without Grade 2 effects was significant, both for rectum only and rectum PRVs (P-value range: 0.02-0.04), with generally lower P-values for the PRVs. In the logistic regression, the fractional DVH variables (i.e. volumes) were significantly related to toxicity, with approximately 2-3 times as many significant dose levels for the PRVs as for rectum only. E.g. with wide anterior and posterior margins (16 and 11 mm, respectively) the relation was significant at 26 different dose levels (6-7, 13-14, 35-43, 60-71 and 73 Gy), compared to nine levels (38-40, 43-44 and 71-74 Gy) for rectum only. EUDs were significantly different for patients with and without Grade 2 effects both for rectum only and the PRVs (95% confidence interval for EUD increase with Grade 2 effects: 0.1-3.1 Gy). CONCLUSIONS: All statistical methods applied indicated a small, but definite difference in DVH parameters between patients with versus those without Grade 2 effects. The difference was most pronounced when margins of 16 mm anterior and 11 mm posterior were applied. PMID- 15878631 TI - CHART in lung cancer: economic evaluation and incentives for implementation. AB - BACKGROUND AND PURPOSE: To investigate the financial consequences and the impact on daily implementation of CHART in lung cancer. PATIENTS AND METHODS: A cost effectiveness and cost-utility analysis were performed using Markov models, comparing the early and delayed costs and effects of CHART for NSCLC over a 4 year time span from a societal viewpoint. The outcome estimates were based on the CHART literature, the cost estimates on the standard practice of the Leuven University Hospitals, the radiotherapy costs being derived from an activity-based costing (ABC) programme developed in the department. RESULTS: The additional societal cost per life-year gained was 9164 Euro, the incremental cost per quality-adjusted life-year 11,576 Euro. Sensitivity analyses confirmed the robustness of these results, the incremental cost-utility ratio remaining well under 20,000 Euro/QALY in all tested circumstances. The threshold analyses found the results of the study to be sensitive to the cost of CHART and to the quality of life after treatment. More specifically, standard treatment would become the optimal treatment if CHART would have a higher cost or would result in more long term side effects. CONCLUSION: CHART should not be denied to patients with NSCLC on the basis of clinical or economic arguments. Other factors such as socio economical, institutional, practical departmental and physician-bound barriers most probably explain the lack of implementation into daily practice. PMID- 15878632 TI - Nonviral HVJ (hemagglutinating virus of Japan) liposome-mediated retrograde gene transfer of human hepatocyte growth factor into rat nervous system promotes functional and histological recovery of the crushed nerve. AB - Hepatocyte growth factor (HGF) is well known to be involved in many biological functions, such as organ regeneration and angiogenesis, and to exert neurotrophic effects on motor, sensory, and parasympathetic neurons. In this study, we gave repeated intramuscular injections of the human HGF gene, using nonviral HVJ (hemagglutinating virus of Japan) liposome method, to examine whether transfection of the rat nervous system with this gene is able to exert neurotrophic effects facilitating recovery of a crushed nerve. The expression of HGF protein and HGF mRNA indicated that gene transfer into the nervous system did occur via retrograde axonal transport. At 4 weeks after crush, electrophysiological examination of the crushed nerve showed a significantly shorter mean latency and a significantly greater mean maximum M-wave amplitude with repeated injections of HGF gene. Furthermore, histological findings showed that the mean diameter of the axons, the axon number and the axon population were significantly larger in the group with repeated injections of HGF gene. The above results show that repeated human HGF gene transfer into the rat nervous system is able to promote crushed-nerve recovery, both electrophysiologically and histologically, and suggest that HGF gene transfer has potential for the treatment of crushed nerve. PMID- 15878633 TI - Galactosylated polyethylenimine-graft-poly(vinyl pyrrolidone) as a hepatocyte targeting gene carrier. AB - Polyethylenimine (PEI) has been used for the gene delivery system in vitro and in vivo since it has high transfection efficiency owing to proton buffer capacity. However, the use of PEI for gene delivery is limited due to cytotoxicity, non specificity and unnecessary interaction with serum components. To overcome cytotoxicity and non-specificity, PEI was coupled with poly(vinyl pyrrolidone) (PVP) as the hydrophilic group to reduce cytotoxicity and lactose bearing galactose group for hepatocyte targeting. The galactosylated-PEI-graft-PVP (GPP) was complexed with DNA, and GPP/DNA complexes were characterized. GPP showed good DNA binding ability, high protection of DNA from nuclease attack. The sizes of DNA complexes show tendency to decrease with an increase of charge ratio and had a minimum value around 59 nm at the charge ratio of 40 for the GPP-1/DNA complex (PVP content: 4.1 mol%). The GPP showed low cytotoxicity. And GPP/DNA complexes were mediated by asialoglycoprotein receptors (ASGP-R)-mediated endocytosis. Also, the transfection efficiency of GPP-1/DNA complex at charge ratio of 40 in the HepG2 was higher than that of PEI/DNA one. PMID- 15878634 TI - Bacterial ghosts as antigen delivery vehicles. AB - The bacterial ghost system is a novel vaccine delivery system unusual in that it combines excellent natural intrinsic adjuvant properties with versatile carrier functions for foreign antigens. The efficient tropism of bacterial ghosts (BG) for antigen presenting cells promotes the generation of both cellular and humoral responses to heterologous antigens and carrier envelope structures. The simplicity of both BG production and packaging of (multiple) target antigens makes them particularly suitable for use as combination vaccines. Further advantages of BG vaccines include a long shelf-life without the need of cold chain storage due to their freeze-dried status, they are safe as they do not involve host DNA or live organisms, they exhibit improved potency with regard to target antigens compared to conventional approaches, they are versatile with regards to DNA or protein antigen choice and size, and as a delivery system they offer high bioavailability. PMID- 15878635 TI - Demonstration plasma gasification/vitrification system for effective hazardous waste treatment. AB - Plasma gasification/vitrification is a technologically advanced and environmentally friendly method of disposing of waste, converting it to commercially usable by-products. This process is a drastic non-incineration thermal process, which uses extremely high temperatures in an oxygen-starved environment to completely decompose input waste material into very simple molecules. The intense and versatile heat generation capabilities of plasma technology enable a plasma gasification/vitrification facility to treat a large number of waste streams in a safe and reliable manner. The by-products of the process are a combustible gas and an inert slag. Plasma gasification consistently exhibits much lower environmental levels for both air emissions and slag leachate toxicity than other thermal technologies. In the framework of a LIFE-Environment project, financed by Directorate General Environment and Viotia Prefecture in Greece, a pilot plasma gasification/vitrification system was designed, constructed and installed in Viotia Region in order to examine the efficiency of this innovative technology in treating industrial hazardous waste. The pilot plant, which was designed to treat up to 50kg waste/h, has two main sections: (i) the furnace and its related equipment and (ii) the off-gas treatment system, including the secondary combustion chamber, quench and scrubber. PMID- 15878636 TI - Hindlimb stepping movements in complete spinal rats induced by epidural spinal cord stimulation. AB - The locomotor ability of the spinal cord of adult rats deprived of brain control was tested by epidural spinal cord stimulation. The studies were performed on six rats that had a complete spinal cord transection (T7-T9) and epidural electrode implantations 2-3 weeks before testing was initiated. The stimulating epidural electrodes were implanted at the T12-L6 spinal segments. Epidural electrical stimulation of the dorsal surface of the spinal cord at frequencies between 1 and 50 Hz and intensities between 1 and 10 V without any pharmacological facilitation was used. Stimulation at each of the lumbar spinal cord segments elicited some rhythmic activity in the hindlimbs. However, stimulation at most segmental levels usually evoked activity in only one leg and was maintained for short periods of time (< 10s). Bilateral hindlimb locomotor activity was evoked most often with epidural stimulation at 40-50 Hz applied at the L2 segment. A necessary condition for initiation of locomotor activity was providing a specific amount (at least 5%) of body weight support. Therefore, the rat spinal cord isolated from brain control is capable of producing bilateral stepping patterns induced most readily by epidural stimulation applied at the L2 spinal segment. Furthermore, the induced stepping patterns were dependent on sensory feedback associated with weight bearing. PMID- 15878637 TI - Evidence for lectin activity of a plant receptor-like protein kinase by application of neoglycoproteins and bioinformatic algorithms. AB - Detection of genes for putative receptor-like protein kinases, which contain an extracellular domain related to leguminous lectins, in plant genomes inspired the hypothesis that this part acts as sensor. Initial support for this concept came from proof for protein kinase activity. The next step, focusing on the protein of lombardy poplar (Populus nigra var. italica), is scrutiny for lectin activity. Consequently, we first pinpointed sets of high-scoring sequence pairs by extensive databank search. The calculations resulted in P-values in the range from 10(-14) to 10(-18) exclusively for leguminous lectins, the Pterocarpus angolensis agglutinin being front runner with P=3 x 10(-18) and thus most suitable template for modeling. The superimposition of the two folds gave notable similarity in the region responsible for binding carbohydrate and Ca(2+)/Mn(2+) ions. Binding activity toward carbohydrates was detected by assaying a panel of (neo)glycoproteins as polyvalent probes, especially for alpha-l-rhamnose and glycans of asialofetuin. It was strictly dependent on Ca(2+)-ions, enhanced by Mn(2+)-ions and reached a K(D)-value of 34.3 nM for the neoglycoprotein with rhamnose as ligand. These results give further research direction to define physiological ligands, plant/bacterial rhamnose-containing saccharides and rhamnose-mimetic glycans or peptides being potential candidates. PMID- 15878638 TI - Reactivity towards singlet oxygen of propofol inside liposomes and neuronal cells. AB - Singlet oxygen (1O2), a reactive oxygen species, has been found to be implicated in many cellular events and pathological disorders. Herein, we investigated the reactivity of 1O2 towards the anaesthetic agent propofol (PPF) encapsulated within DMPC liposomes. By time resolved luminescence, the rate constant of 1O2 quenching by PPF was evaluated, depending on the location of the sensitizer, with following values: 1.35+/-0.05x10(7) M(-1) s(-1) for deuteroporphyrin (as embedded source) and 0.8+/-0.04x10(7) M(-1) s(-1) for uroporphyrin (as external source), respectively. The nature of the oxidation product, resulting from the reaction of 1O2 with PPF, was determined using absorption and HPLC techniques. Finally, the in vitro protective effect of PPF towards the 1O2-induced neuronal cell toxicity was evaluated in terms of cell viability. PMID- 15878639 TI - Identification and immunohistochemical localization of Sulfotransferase 2B1b (SULT2B1b) in human lung. AB - Sulfotransferase 2B1b (SULT2B1b) is a member of SULT 2 gene family. SULT2B1a and SULT2B1b are transcribed from the same gene using different transcriptional start sites and contain different first exons as the result of alternative splicing. SULT2B1a and SULT2B1b are 350 and 365 AA in length, respectively. Northern blot analysis and SULT2B1 isoform specific RT-PCR detected only SULT2B1b message in human lung tissue. Immunoblot analysis of human lung tissue with a specific rabbit anti-SULT2B1 polyclonal antibody detected only SULT2B1b immunoreactive protein in human lung cytosol. Immunoprecipitation and MALDI mass spectroscopic analysis verified that the immunoreactive protein was SULT2B1b. Immunohistochemical localization of SULT2B1b in human tissues showed expression in the cytoplasm of ciliated columnar or cuboidal epithelial cells in terminal bronchia. No staining was observed in alveolar cells. SULT2B1b is selective for the sulfation of 3beta-hydroxysteroids such as dehydroepiandrosterone and pregnenolone as well as cholesterol. The presence of SULT2B1b in lung tissues suggests a role in the regulation of local steroid hormone synthesis and metabolism. PMID- 15878640 TI - New insights in the visualization of membrane permeabilization and DNA/membrane interaction of cells submitted to electric pulses. AB - Electropermeabilization designates the use of electric pulses to overcome the barrier of the cell membrane. This physical method is used to transfer anticancer drugs or genes into living cells. Its mechanism remains to be elucidated. A position-dependent modulation of the membrane potential difference is induced, leading to a transient and reversible local membrane alteration. Electropermeabilization allows a fast exchange of small hydrophilic molecules across the membrane. It occurs at the positions of the cell facing the two electrodes on an asymmetrical way. In the case of DNA transfer, a complex process is present, involving a key step of electrophoretically driven association of DNA only with the destabilized membrane facing the cathode. We report here at the membrane level, by using fluorescence microscopy, the visualization of the effect of the polarity and the orientation of electric pulses on membrane permeabilization and gene transfer. Membrane permeabilization depends on electric field orientation. Moreover, at a given electric field orientation, it becomes symmetrical for pulses of reversed polarities. The area of cell membrane where DNA interacts is increased by applying electric pulses with different orientations and polarities, leading to an increase in gene expression. Interestingly, under reversed polarity conditions, part of the DNA associated with the membrane can be removed, showing some evidence for two states of DNA in interaction with the membrane: DNA reversibly associated and DNA irreversibly inserted. PMID- 15878641 TI - Fos expression in the suprachiasmatic nucleus in response to light stimulation in a solitary and social species of African mole-rat (family Bathyergidae). AB - Mole-rats are strictly subterranean rodents that are rarely exposed to environmental light. They are well adapted to their environment and have reduced eyes and a severely regressed visual system. It has been shown, however, that mole-rats do exhibit endogenous circadian rhythms that can be entrained, suggesting an intact and functional circadian system. To determine whether light is the entraining agent in these animals, Fos expression in response to light pulses at different circadian times was investigated to obtain phase response curves. Light is integrated effectively in the suprachiasmatic nucleus of the Cape mole-rat (Georychus capensis), and Fos expression is gated according to the phase of the circadian clock. The Fos response in the Cape mole-rat was comparable to that of aboveground rodents. In contrast, the highveld mole-rat (Cryptomys hottentotus pretoriae) was less sensitive to light and did not show a selective Fos response according to the phase of the circadian cycle. Social species appear to be less sensitive to light than their solitary counterparts, which compares well with results from locomotor activity studies. PMID- 15878642 TI - Protein kinase Calpha modulates depolarizaton-evoked changes of intracellular Ca2+ concentration in a rat pheochromocytoma cell line. AB - Conventional protein kinase C (cPKC) isoforms are activated by a coincident rise in cytosolic Ca(2+) and membrane-bound diacylglycerol. In excitable cells, cPKC may be activated by Ca(2+) influx through voltage-gated Ca(2+) channels (VGCC). cPKCs, in turn, are known to modulate the activity of VGCC. We examined whether PKCalpha, a cPKC, could be activated by depolarization in a neuroendocrine cell line and whether activation occurred on a time scale that modulated the depolarization-evoked intracellular Ca(2+) concentration ([Ca(2+)](i)) signal. Pheochromocytoma cells (PC12 cells) were transfected with wild-type and mutant forms of PKCalpha labeled with yellow fluorescent protein to monitor kinase translocation. Simultaneously, [Ca(2+)](i) changes were monitored with fura-2. Two point mutations that render PKCalpha inactive, D187A in the Ca(2+) binding site and K368R in the ATP binding site, significantly prolonged the time-to-peak of the depolarization-evoked [Ca(2+)](i) signal. A mutation that modulates membrane insertion (W58G) and two mutations of an autophosphorylation site (S657A, S657E) had no effect on the kinetics of the [Ca(2+)](i) signal. We conclude that in PC12 cells, Ca(2+) entry through VGCC rapidly activates PKCalpha, and that PKCalpha can modulate the Ca(2+) signal on a physiologically relevant time scale. Point mutations of PKCalpha can be used as specific and potent modulators of the PKC signaling pathway. PMID- 15878643 TI - Involvement of extracellular signal-regulated kinases 1/2 and (phosphoinositide 3 kinase)/Akt signal pathways in acquired resistance against neurotoxin of 6 hydroxydopamine in SH-SY5Y cells following cell-cell interaction with astrocytes. AB - Glial cells interact with neurons and play important roles in the development, differentiation, maintenance and repair of the nervous system. Human neuroblastoma cells (SH-SY5Y) became dramatically resistant to neurotoxin 6 hydroxydopamine (6-OHDA), when co-cultured with mouse astrocytes. In order to further delineate the molecular mechanism involved in the neuroprotection in this selective cell-cell interaction, we assessed the activation of two signal pathways, namely, the MAP kinases (extracellular signal-regulated kinases, ERK1/2) and phosphoinositide 3-kinase (PI3-K)/Akt signal pathways in response to 6-OHDA insult and subsequent neuronal survival. Western blot revealed that 6-OHDA significantly increased the phosphorylation of ERK1/2 and Akt in mono-cultured SH SY5Y cells. However, the increase in ERK1/2 in SH-SY5Y cells after co-cultured with astrocytes occurred as early as 3 h after 6-OHDA treatment in oppose to the increase after 12 h in monocultures. The phosphorylation of Akt in the co cultured SH-SY5Y cells was much pronounced 3 h after 6-OHDA treatment compared with that in the mono-cultured cells. The anti-apoptotic protein bcl-2 was also increased in the co-cultured SH-SY5Y cells 3 h after treatment with 6-OHDA. Selective inhibitor of PI3-K/Akt signal pathway blocked the acquired resistance to 6-OHDA in SH-SY5Y cells following interaction with astrocytes. Inhibition of ERK1/2 signal pathway did not affect the cell survival. Our data suggest that PI3 K/Akt signal pathway, but not ERK1/2, is involved the acquired resistance in SH SY5Y cells following cell-cell interaction with astrocytes against the neurotoxic 6-OHDA insult. PMID- 15878644 TI - An antidepressant mechanism of desipramine is to decrease tumor necrosis factor alpha production culminating in increases in noradrenergic neurotransmission. AB - The monoamine theory of depression proposes decreased bioavailability of monoamines, such as norepinephrine (NE), as the underlying cause of depression. Thus, the antidepressant efficacy of NE-reuptake inhibitors such as desipramine is attributed to increases in synaptic concentrations of NE. The time difference between inhibition of reuptake and therapeutic efficacy, however, argues against this being the primary mechanism. If desipramine elicits its therapeutic efficacy by increasing NE release, in turn, increasing activation of the alpha(2) adrenergic autoinhibitory receptor, then mimicking this increase with an exogenous agonist (clonidine) should support or even enhance the efficacy of the antidepressant. Intriguingly, simultaneous administration of clonidine with desipramine prevented the cellular and behavioral effects elicited by desipramine alone, in both acute and chronic administration paradigms. These results suggest the involvement of additional factor(s) in the mechanism of antidepressant action of this drug. Desipramine administration results in a virtual ablation of neuron derived tumor necrosis factor-alpha (TNF), thus implicating an essential role of TNF in the therapeutic efficacy of this antidepressant. Additionally, following chronic administration of desipramine, TNF-regulation of NE release is transformed, from inhibition to facilitation. Here, we demonstrate that a transformation in TNF-regulation of NE release in the brain is a key element in the efficacy of this antidepressant. Interestingly, an increase in neurotransmission prior to the antidepressant's effect on TNF production prevents the efficacy of the antidepressant drug. Thus, the efficacy of desipramine is due to decreased levels of TNF in the brain induced by this drug, ultimately modifying noradrenergic neurotransmission. PMID- 15878645 TI - Mucosal vaccination delays or prevents prion infection via an oral route. AB - In recent years major outbreaks of prion disease linked to oral exposure of the prion agent have occurred in animal and human populations. These disorders are associated with a conformational change of a normal protein, PrP(C) (prion protein cellular), to a toxic and infectious form, PrP(Sc) (prion protein scrapie). None of the prionoses currently have an effective treatment. A limited number of active immunization approaches have been shown to slightly prolong the incubation period of prion infection. Active immunization in wild-type animals is hampered by auto-tolerance to PrP and potential toxicity. Here we report that mucosal vaccination with an attenuated Salmonella vaccine strain expressing the mouse PrP, is effective at overcoming tolerance to PrP and leads to a significant delay or prevention of prion disease in mice later exposed orally to the 139A scrapie strain. This mucosal vaccine induced gut anti-PrP immunoglobulin (Ig)A and systemic anti-PrP IgG. No toxicity was evident with this vaccination approach. This promising finding suggests that mucosal vaccination may be a useful method for overcoming tolerance to PrP and preventing prion infection among animal and potentially human populations at risk. PMID- 15878646 TI - The distribution of inhibitory and excitatory synapses on single, reconstructed jaw-opening motoneurons in the cat. AB - In a previous study, we reported that the distribution of inhibitory input, in contrast to excitatory input, decreased somatofugally along dendrites of cat jaw closing alpha-motoneurons [J Comp Neurol 414 (1999) 454]. The present study examined the distribution of GABA, glycine, and glutamate immunopositive boutons covering horseradish peroxidase-labeled cat jaw-opening motoneurons. The motoneurons were divided into four compartments: the soma, and primary, intermediate, and distal dendrites. Ninety-seven percent of the total number of studied boutons had immunoreactivity for at least one of the three amino acids. The proportion of boutons immunoreactive for GABA and/or glycine was lower than the proportion of boutons immunoreactive for glutamate. Boutons immunoreactive to glycine alone were more numerous than boutons double-labeled for GABA and glycine, which, in turn, occurred more frequently than boutons immunoreactive to GABA alone. The percentage synaptic covering (proportion of membrane covered by synaptic boutons) of the putatively excitatory (glutamate containing) and putatively inhibitory (GABA and/or glycine containing) boutons decreased somatofugally along the dendrites. Such systematic variations were not seen in the packing density (number of boutons per 100 microm(2)); the packing density showed a distinct drop between the soma and primary dendrites but did not differ significantly among the three dendritic compartments. Overall, the packing density was slightly higher for the putatively excitatory boutons than for the inhibitory ones. When taken together with previous analyses of jaw-closing alpha motoneurons the present data on jaw-opening alpha-motoneurons indicate that the two types of neuron differ in regard to the nature of synaptic integration in the dendritic tree. PMID- 15878647 TI - The Drosophila black enigma: the molecular and behavioural characterization of the black1 mutant allele. AB - The cuticular melanization phenotype of black flies is rescued by beta-alanine, but beta-alanine production, by aspartate decarboxylation, was reported to be normal in assays of black mutants, and although black/Dgad2 is expressed in the lamina, the first optic ganglion, no electroretinogram (ERG) or other visual defect has been demonstrated in black flies. The purpose of this study was to investigate the black gene, and protein, in black(1) mutants of Drosophila melanogaster in order to resolve the apparent paradox of the black phenotype. Using black(1) mutant flies we show that (1) aspartate decarboxylase activity is significantly reduced in adults and at puparium formation, consistent with defects in cuticular and non-cuticular processes, (2) that the black(1) mutation is a frameshift, and black(1) flies are nulls for the black/DGAD2 protein, and (3) that behavioural experiments using Buridan's paradigm, demonstrate that black responds abnormally to visual cues. No ERG, or target recognition defects can be demonstrated suggesting a problem with higher order visual functions in black mutants. PMID- 15878648 TI - MS-KIF18A, new kinesin; structure and cellular expression. AB - The present study describes the cloning and molecular analysis of a new gene, MS KIF18A, a member of the kinesin family. MS-KIF18A was cloned from a marrow stromal cells expression library. Transcripts derived from this gene were also detected in testis and trachea, although they differed from the stroma mesenchymal cell transcript in the open reading frame (ORF) as well as in the untranslated regions (UTRs). The existence of various transcripts suggests alternative regulation of gene expression and defines tissue specific function of the new kinesin. The cDNA from the marrow stroma, MS-KIF18A, encodes a predicted protein of 898 amino acids with a molecular weight of 100 kDa. Kinesins are motor proteins that consist of a motor domain with microtubule-binding and ATPase sites, a coiled coil region and a cargo-binding domain. Examination of a three dimensional model of the MS-KIF18A motor domain suggested that this protein associates with microtubules, which was confirmed by immunofluorescence (IF) experiments in stromal cells. PMID- 15878649 TI - Allophylus serratus: a plant with potential anti-ulcerogenic activity. AB - Allophylus serratus is known to possess various therapeutic properties. We evaluated the anti-ulcerogenic property of crude ethanolic extract of Allophylus serratus (AS) in different ulcer models in Sprague-Dawley rats. The extract at 400 mg/kg body weight, once daily, orally has a significant effect in cold restraint (CRU, 2 h cold restraint stress), aspirin (ASA, 150 mg/kg body weight, orally), alcohol (AL, 1 ml/200 gm of absolute alcohol) and pyloric ligation (PL, 4h ligation) induced gastric ulcer models as it showed protection index of 71.28, 62.50, 90.84 and 64.29% protection, respectively whereas, standard drug omeprazole (OMZ, 10mg/kg body weight) has shown protection index of 85.70, 74.99 and 74.99 in CRU, ASA and PL model respectively. Sucralfate (SUC, 500 mg/kg body weight) as a standard drug in AL model has 93.20% protection. Furthermore, AS has significantly decreased the free acidity (72.41%), total acidity (47.97%) and peptic activity (24.59%), respectively as well as has significantly increased the mucus secretion (29.41%). Conclusively the ulcer protective effect of AS may be due to its anti-secretory along with cytoprotective mechanism. PMID- 15878650 TI - Immunological responses to bone soluble proteins in recipients of bone allografts. AB - The significance of an immune response in complications of bone allograft procedures is not well understood. This study evaluates the immunological reaction to bone allografts in either cortical or cancellous form. Serological responses from osteoarthritis (OA) patients and normal individuals to soluble proteins extracted from allografts were assessed using Western blotting and ELISA techniques. A large number of patients expressed antibodies to bone proteins extracted from the washed bone as compared with normal controls. Antibodies were present in patients without the use of bone allografts, indicating that OA patients develop antibodies to bone soluble proteins. However, patients receiving allografts exhibited an increased immune reactivity against multiple bone proteins when compared with non-grafted patients. Protein characterization of the immunoreactive proteins revealed that the majority of antigenic targets were fragments of various collagen molecules. The data suggests that OA patients develop antibodies to bone soluble proteins prior to surgery, and these antibodies increase after revision surgery utilizing bone allografts. These findings support the implications that various collagen molecules as well as their degraded fragments represent potential immunogenic proteins within bone allografts. The removal of these antigenic proteins from the allografts prior to surgery may alleviate this immunological reactivity and improve graft outcome. PMID- 15878651 TI - Altered gene expression in human hepatoma HepG2 cells exposed to low-level 2,4 dichlorophenoxyacetic acid and potassium nitrate. AB - 2,4-dichlorophenoxyacetic acid (2,4-D) and nitrate are agricultural contaminants found in rural ground water. It is not known whether levels found in groundwater pose a human or environmental health risk, nor is the mechanism of toxicity at the molecular/cellular level understood. This study focused on determining whether 2,4-D or nitrate at environmentally realistic levels elicit gene expression changes in exposed cells. cDNA microarray technology was used to determine the impact of 2,4-D and nitrate in an in vitro model of exposure. Human hepatoma HepG2 cells were incubated with 2,4-D or nitrate alone for 24 h. Cell viability (neutral red assay) and proliferation (BrdU incorporation) were assessed following exposure. Total RNA from treated and control cells were isolated, reverse transcribed and reciprocal labelled with Cy3 or Cy5 dyes, and hybridized to a human cDNA microarray. The hybridized microarray chips were scanned, quantified and analyzed to identify genes affected by 2,4-D or nitrate exposure based on a two-fold increase or decrease in gene expression and reproducibility (affected in three or more treatments). Following filtering, normalization and hierarchical clustering initial data indicate that numerous genes were found to be commonly expressed in at least three or more treatments of 2,4-D or nitrate tested. The affected genes indicate that HepG2 cells respond to environmental, low-level exposure and produce a cellular response that is associated with alterations in the expression of many genes. The affected genes were characterized as stress response, cell cycle control, immunological and DNA repair genes. These findings serve to highlight new pathway(s) in which to further probe the effects of environmental levels of 2,4-D and nitrate. PMID- 15878652 TI - Genetic polymorphisms of toll-like receptor 9 influence the immune response to CpG and contribute to hyper-IgM in primary biliary cirrhosis. AB - The serum hallmark of primary biliary cirrhosis (PBC) is the presence of anti mitochondrial antibodies (AMA), found in 95% of patients. However, nearly every patient with PBC, including those who are AMA-negative, has an elevation in serum IgM. This hyper-IgM is neither representative of other Ig isoforms, nor is due to the levels of AMA. In fact, we have recently reported that the hyper-IgM is an innate immune response and can be induced with CpG-B with concurrent up regulation of toll-like receptor 9 (TLR9). Based on these observations, we performed a two-tier study. First, we quantitated TLR9 genotypes in patients with PBC and controls and correlated these data with the B cell response to CpG-B. Second, based on these data, we performed an extensive TLR9 genotyping in a large cohort of patients and controls. We report herein that the 2848 AA TLR9 genotype is associated with enhanced gene expression and higher frequency of intracellular IgM(+) B cells following CpG stimulation. Interestingly, however, despite the functional association, there is no difference in the distribution of TLR9 genotypes between patients and controls. Our data emphasize the importance of dissecting the innate immune response in PBC. PMID- 15878653 TI - Extended-spectrum beta-lactamases in Shigella flexneri from Argentina: first report of TOHO-1 outside Japan. AB - A 9-year nation-wide survey of the presence of extended-spectrum beta-lactamases (ESBLs) in Shigella flexneri is described. Ten of 9033 (0.1%) isolates produced ESBLs, which were characterized by isoelectric focusing, PCR and DNA sequencing. These were CTX-M-2 (five isolates), TOHO-1 (one isolate), SHV-2 (two isolates) and PER-2 (two isolates, the first report in S. flexneri world wide). The emergence of each ESBL type in S. flexneri was not restricted to a particular region of Argentina. TOHO-1 showed a more basic isoelectric point (8.4) than that previously found (7.8) and its encoding gene (bla(TOHO-1a)) harboured a silent change, G825A, relative to the reported bla(TOHO-1). All the ESBL-encoding genes were transferred to Escherichia coli by conjugation. PFGE analysis indicated that the 10 ESBL-producing S. flexneri isolates were subtypes of a unique clone. PMID- 15878654 TI - Reversal of the late asthmatic response increases exhaled nitric oxide. PMID- 15878655 TI - Continuous oxygen monitoring--a better way to prescribe long-term oxygen therapy. AB - STUDY OBJECTIVE: The purpose of this study was to determine if an oxygen prescription based on continuous oximetry monitoring, would result in an increased percentage of time spent within an SpO2 level between 88% and 92%. METHODS: We conducted a prospective, cohort study in an outpatient, pulmonary setting in a tertiary care referral center, on 17 patients with stable chronic obstructive pulmonary disease (COPD) who had previously been prescribed long-term oxygen therapy. The patients were monitored for approximately 24h with a portable oximeter that recorded SpO2 and EKG readings. During the initial 24h of monitoring, the patients were on their previous oxygen prescription. Their oxygen prescription was then altered based on a predetermined protocol described below. The patients were then monitored for an additional 24h. RESULTS: Sixteen patients completed the study. Based on the initial continuous oximetry, the median oxygen prescription was reduced from 2.5 to 1.2 L/min (P < 0.001). The oxygen prescription during exercise, rest, and sleep decreased from 3.0 +/- 0.9 to 1.8 +/- 1.3 L/min (P < 0.001), 2.2+/-0.4 to 0.8 +/- 0.7 L/min (P < 0.001), and 2.2 +/ 0.4 to 0.9 +/- 0.8 L/min (P < 0.001) respectively. After the oxygen prescription was adjusted, the percentage of the time that the SpO2 was between 88 and 92% increased from 24.8 +/- 21.7% to 52.8 +/- 25.0% (P = 0.001), but the percentage of time that the SpO2 was below 88% did not change significantly (2.8 +/- 6.1% to 4.3 +/- 8.1%). CONCLUSIONS: This study demonstrates that an oxygen prescription based on continuous oximetry monitoring results in (1) a significant increase in the percentage of time that the SpO2 is between 88% and 92%, (2) a significant decrease in the amount of oxygen prescribed, (3) a slight increase in the amount of time that the SpO2 is below 88% that was not statistically significant. PMID- 15878656 TI - Androgen action on hepatic vitellogenin synthesis in the eel, Anguilla japonica is suppressed by an androgen receptor antagonist. AB - Involvement of additional hormones other than estrogen in the control of vitellogenin (Vg) synthesis has been suggested in fish. However, no satisfactory explanation on the mechanism of the action of these hormones has been reported. In this study, we have exploited the possibility of androgen receptor mediation during the androgen action on the pathway of Vg synthesis. Hepatocytes were prepared from sexually immature Japanese eel Anguilla japonica and treated with estradiol-17beta (E2), 17alpha-methyltestosterone (MT), growth hormone, tamoxifen or flutamide, or in combination of these. Spent culture media were analysed by SDS-PAGE for Vg detection. Results from the chemical treatments demonstrated the necessity of E2 as the primary factor for Vg synthesis and requirement of additional hormones for the full expression of Vg. The effects of E2 and MT were effectively blocked by tamoxifen, an estrogen receptor antagonist and flutamide, an androgen receptor antagonist, respectively, indicating ER-mediated estrogen action and AR-mediated androgen action on Vg synthesis in this species. PMID- 15878657 TI - Hydrothermal upgrading of biomass: effect of K2CO3 concentration and biomass/water ratio on products distribution. AB - Catalytic hydrothermal treatment of wood biomass was performed at 280 degrees C for 15 min in the presence of K2CO3 with different concentrations and biomass/water ratio (thermal). Oil products were extracted from both liquid and solid portion by different solvents and analyzed them individually. The biomass to water ratio has an important effect on product distribution and composition of oil products. Oil 1 (ether extract) with K2CO3 contained mainly phenolic compounds. Benzenediol derivatives were observed with 0.94 M K2CO3 concentration and they were not formed at lower concentrations (0.235 and 0.47 M). The decrease of solid residue was achieved to 4% with 0.94 M K2CO3 at 280 degrees C for 15 min. The volatility distribution of hydrocarbons (ether extract) were characterized by using C-NP gram. The distribution of oxygenated hydrocarbons changed depending upon the biomass to water ratio and concentration of K2CO3 solution. PMID- 15878658 TI - Effect of salinity and temperature on germination, growth and ion relations of Panicum turgidum Forssk. AB - Seed germination of Panicum turgidum was significantly affected by salinity levels, temperature and their interaction. Maximum germination was noted in the lowest saline media (25-50 mM) and distilled water at the temperature of 15-25 degrees C and 20-30 degrees C. Seeds germination was substantially delayed and reduced with an increase in NaCl to levels above 50mM. This trend was much pronounced under high levels of NaCl and incubation temperature. Low levels of NaCl (25-50 mM) stimulated shoot and root dry weights of P. turgidum seedlings. However, the highest NaCl levels (>100 mM) resulted in a significant decrease in shoot, root and total dry weights of seedlings. Intermediate degrees of temperature, 15-25 and 20-30 degrees C, resulted in a significant increase in biomass accumulation. The Na+ concentration in shoots and roots significantly increased as NaCl concentration increased. The K+ concentration in roots and K/Na ratio in shoots and roots was significantly reduced as salinity concentration increased. The K/Na ratio was greatly affected by higher NaCl concentration and incubation temperatures. PMID- 15878659 TI - Non-competitive inhibitors of metabotropic glutamate receptor 5 (mGluR5). AB - Based on a pharmacophore alignment on known non-competitive mGluR5 inhibitors applying 4SCan technology, a new lead series was identified and further structurally investigated. K(i)'s as low as around 100 nM were achieved. PMID- 15878660 TI - Redox-responsive and calcium-dependent switching of glycosyldisulfide interactions with Concanavalin A. AB - Glycosyldisulfides can interact efficiently with carbohydrate-binding entities. This has been shown for a range of thiosaccharide dimers when tested for their effects against the lectin Concanavalin A using a modified quartz crystal microbalance-technique. Contrary to the thiosaccharide monomers, showing no significant binding up to 10 mM, several of the dimers showed IC(50)-values in the low millimolar range. Three of the glycosyldisulfides tested also displayed very high positive apparent cooperativity effects that were found to be both calcium-dependent and redox-responsive. PMID- 15878661 TI - 2D RNA-QSAR: assigning ACC oxidase family membership with stochastic molecular descriptors; isolation and prediction of a sequence from Psidium guajava L. AB - Quantitative structure-activity relationship (QSAR) techniques for small molecules could be applied to nucleic acids. Unfortunately, almost all molecular descriptors are more successful at encoding branching information than sequences and/or cannot be back-projected. A solution for scaling the QSAR problem up to RNA may be to transform sequences into secondary structures first. Our group has used Markovian negentropies as molecular descriptors for drug design with preliminary results in bioinformatics [Bioinformatics 2003, 19, 2079]. However, RNA-QSAR studies on RNA molecules have not been described to date. Novel Markovian negentropies have been introduced here as molecular descriptors for 2D RNA structures. An RNA-QSAR study of the ACC proteins from different plants has been carried out. The QSAR recognizes 19/20 sequences (95.0%) within the ACC family and 12/17 (70.6%) of the control group sequences. The model has a high Matthews' regression coefficient (C = 0.68). Overall cross-validation average accuracies were 14 out of 15 for ACC sequences (93.3%) and 10 out of 13 for control sequences (76.9%). Finally, ACC oxidase family membership was assigned to a new sequence isolated for the first time in this work from Psidium guajava L. A backprojection map for this sequence identifies the left stem (40%) and the main stem (45%) as highly important substructures. Results of an nBLAST experiment are consistent with this finding and indicate a high conservation score (>70) for left stem and main stem; whereas major loop, right stem, cap and major loop right half were hardly conserved. PMID- 15878662 TI - Synthesis and binding affinity of novel 3-aminoethyl-1-tetralones, potential atypical antipsychotics. AB - A series of 3-aminoethyl-1-tetralones, conformationally constrained higher homologues of haloperidol (standard for typical antipsychotic profile), have been obtained by a four-step route from valerolactone. Their binding affinities at dopamine D(2) and serotonin 5-HT2A and 5-HT2C receptors were determined, showing in some cases an atypical antipsychotic profile. PMID- 15878663 TI - Mechanism of biochemical action of substituted 4-methylcoumarins. Part 11: Comparison of the specificities of acetoxy derivatives of 4-methylcoumarin and 4 phenylcoumarin to acetoxycoumarins: protein transacetylase. AB - Our earlier observations led to the identification of a microsomal enzyme termed as acetoxy drug: protein transacetylase (TAase) catalyzing the transfer of acetyl groups from acetylated polyphenols to the receptor proteins. TAase was conveniently assayed by the irreversible inhibition of cytosolic glutathione S transferase (GST) by the model acetoxycoumarin, 7,8-diacetoxy-4-methylcoumarin (1). The specificities of the acetoxy group on the benzenoid ring and position of the pyran carbonyl group of the coumarin with respect to oxygen heteroatom for the catalytic activity of TAase were also reported earlier. In this communication, we have demonstrated that the acetoxy coumarins and acetoxy dihydrocoumarins having a methyl group instead of a phenyl ring at the C-4, when used as the substrates, resulted in enhancement of TAase activity, while the saturation of double bond at C-3 and C-4 position had no effect on TAase activity. A comparison of the optimized structures of 1 and 7,8-diacetoxy-4 phenylcoumarin (2) suggested that the observed influence may be due to out of plane configuration of the phenyl ring at C-4. Further, the TAase-catalyzed activation of NADPH cytochrome c reductase and inhibition of aflatoxin B1 (AFB1) DNA binding by acetoxy 4-phenylcoumarins and dihydrocoumarins were significantly lower as compared to those caused by acetoxy 4-methylcoumarins. PMID- 15878664 TI - Steroid-derived phospholipid scramblases induce exposure of phosphatidylserine on the surface of red blood cells. AB - A series of methyl 7alpha,12alpha-bis(phenylurea) cholate derivatives with different cationic substituents at the 3alpha-position were prepared and evaluated for an ability to increase the level of endogenous phosphatidylserine (PS) on the surface of red blood cells (erythrocytes). Some of the compounds induced large fractions of erythrocytes to expose sufficient PS to become stained by the protein annexin V-FITC. In addition, the compounds were found to bind PS in homogeneous solution, and to promote the translocation of fluorescent NBD labeled phospholipids across vesicle membranes, which supports the hypothesis that cholate-induced exposure of endogenous PS on the erythrocyte surface is due to the ability of the cationic cholates to promote anionic phospholipid flip flop. PMID- 15878665 TI - Theoretical study revealing the functioning of a novel combination of catalytic motifs in histone deacetylase. AB - Histone deacetylases (HDACs) have recently attracted considerable interest as targets in the treatment of cell proliferative diseases such as cancer. In the present work, the chemical properties of the active site of HDAC were theoretically investigated at a high computational level. Evidence was gathered for a novel catalytic mechanism, which differs from a previous proposal in the native protonation state of the His-Asp dyads, and in the deprotonation of water as a distinct step in the mechanism. PMID- 15878666 TI - Towards new boron carriers for boron neutron capture therapy: metallacarboranes and their nucleoside conjugates. AB - Thymidine conjugates containing metallacarborane, {8-[5-(N(3)-thymidine)-3-oxa pentoxy]-3-cobalt bis(1,2-dicarbollide)}- (5) and {8-[5-(O(4)-thymidine)-3-oxa pentoxy]-3-cobalt bis(1,2-dicarbollide)}- (6) ions and several simple [3-cobalt bis(1,2-dicarbollide)]- ion (1) derivatives have been studied as potential boron carriers for BNCT. Compound 6 and some nonnucleoside derivatives of 1 were not toxic above 100 microM. The partition coefficient for both metallacarborane bearing thymidine conjugates 5 and 6 was more than 500 times higher than that of unmodified nucleoside. The cellular uptake studies showed accumulation of compounds 6 in V79 Chinese hamster cells but not of compound 5. The low toxicity of conjugate type of 6 together with its high partition coefficient suggest that judicially designed derivatives of metallacarboranes can be considered as potential boron carriers for BNCT. PMID- 15878667 TI - Synthesis, antimalarial, antileishmanial, and antimicrobial activities of some 8 quinolinamine analogues. AB - In the present communication, newly synthesized 8-quinolinamines (25-27) related to previously reported 2-tert-butylprimaquine (2) were evaluated for their in vitro antimalarial activity against chloroquine sensitive and resistant Plasmodium falciparum strains, in vivo antimalarial activity against P. berghei infected mice, in vitro antileishmanial activity against Leishmania donovani, in vitro antimicrobial activity against various fungi and bacteria, and cytotoxicity in a panel of mammalian cell lines. No promising cytotoxicities were observed for compounds reported herein. Analogue 25 was found to exhibit curative antimalarial activity at a dose of 25 mg/kg/dayx4 in a P. berghei infected mice model, and produced suppressive activity at a lower dose of 10 mg/kg/dayx4. In vitro antileishmanial activities (IC50 and IC90) comparable to standard drug pentamidine were exhibited by all synthesized 8-quinolinamines 25-27. At the same time, promising antibacterial and antifungal activities were also observed for synthesized compounds against a panel consisting of several bacteria and fungi. PMID- 15878668 TI - New selective acetylcholinesterase inhibitors designed from natural piperidine alkaloids. AB - Five new piperidine alkaloids were designed from natural (-)-3-O-acetyl spectaline and (-)-spectaline that were obtained from the flowers of Senna spectabilis (sin. Cassia spectabilis, Leguminosae). Two semi-synthetic analogues (7 and 9) inhibited rat brain acetylcholinesterase, showing IC50 of 7.32 and 15.1 microM, and were 21 and 9.5 times less potent against rat brain butyrylcholinesterase, respectively. Compound 9 (1mg/kg, i.p.) was fully efficacious in reverting scopolamine-induced amnesia in mice. The two active compounds (7 and 9) did not show overt toxic effects at the doses tested in vivo. PMID- 15878669 TI - Design and synthesis of glycosidase inhibitor 5-amino-1,2,3,4-cyclohexanetetrol derivatives from (-)-vibo-quercitol. AB - In continuation of development of bioactive inositol derivatives, a 1-O-methyl derivative of 5-amino-5-deoxy-L-talo-quercitol was designed and synthesized as an analogue of the strong alpha-fucosidase inhibitor, 5a-carba-alpha-L fucopyranosylamine, the methyl branch being replaced with methoxyl, and demonstrated to be a moderate alpha-fucosidase inhibitor. The present approach provides a possible route to apply alkyl ethers of aminodeoxyinositols as hexopyranose mimics of biological interest. PMID- 15878670 TI - Dolastatin 11 conformations, analogues and pharmacophore. AB - Twenty analogues of the natural antitumor agent dolastatin 11, including majusculamide C, were synthesized and tested for cytotoxicity against human cancer cells and stimulation of actin polymerization. Only analogues containing the 30-membered ring were active. Molecular modeling and NMR evidence showed the low-energy conformations. The amide bonds are all trans except for the one between the Tyr and Val units, which is cis. Since an analogue restricted to negative 2-3-4-5 angles stimulated actin polymerization but was inactive in cells, the binding conformation (most likely the lowest-energy conformation in water) has a negative 2-3-4-5 angle, whereas a conformation with a positive 2-3-4 5 angle (most likely the lowest energy conformation in chloroform) goes through cell walls. The highly active R alcohol from borohydride reduction of dolastatin 11 is a candidate for conversion to prodrugs. PMID- 15878671 TI - Influence of locoregional radiation treatment on tumor necrosis factor-alpha and interleukin-6 in the serum of patients with head and neck cancer. AB - Serum tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 levels in the serum of 34 patients with head and neck cancer (HNC) undergoing locoregional radiotherapy (RT) were examined. The aim of the RT was definitive in 19 and postoperative adjuvant in 15 patients. Serum TNF-alpha and IL-6 levels were recorded before RT and after the completion of the fifth week of RT. The mean TNF alpha levels before and after RT were 28.26 +/- 2.87 and 83.03 +/- 7.47, and the mean IL-6 levels were 61.56 +/- 14.32 and 122.45 +/- 30.66, respectively. The statistical analysis yielded a significant rise in TNF-alpha levels with RT in all patients (p < 0.0001) and also in IL-6 levels in patients treated with postoperative adjuvant RT (p = 0.001). Irradiation is likely to cause an acute phase response, and the cytokines studied may be used to monitor this clinically important response in further trials. PMID- 15878672 TI - Immune-regulation of the apolipoprotein A-I/C-III/A-IV gene cluster in experimental inflammation. AB - Apolipoprotein A-IV is a member of the apo A-I/C-III/A-IV gene cluster. In order to investigate its hypothetical coordinated regulation, an acute phase was induced in pigs by turpentine oil injection. The hepatic expression of the gene cluster as well as the plasma levels of apolipoproteins were monitored at different time periods. Furthermore, the involvement of the inflammatory mediators' interleukins 1 and 6 and tumor necrosis factor in the regulation of this gene cluster was tested in cultured pig hepatocytes, incubated with those mediators and apo A-I/C-III/A-IV gene cluster expression at the mRNA level was measured. In response to turpentine oil-induced inflammation, a decreased hepatic apo A-IV mRNA expression was observed (independent of apo A-I and apo C-III mRNA) not correlating with the plasma protein levels. The distribution of plasma apo A IV experienced a shift from HDL to larger particles. In contrast, the changes in apo A-I and apo C-III mRNA were reflected in their corresponding plasma levels. Addition of cytokines to cultured pig hepatocytes also decreased apo A-IV and apo A-I mRNA levels. All these results show that the down-regulation of apolipoprotein A-I and A-IV messages in the liver may be mediated by interleukin 6 and TNF-alpha. The well-known HDL decrease found in many different acute-phase responses also appears in the pig due to the decreased expression of apolipoprotein A-I and the enlargement of the apolipoprotein A-IV-containing HDL. PMID- 15878673 TI - Imaging the visual autokinetic illusion with fMRI. AB - During fixation of a stationary, dim light-emitting diode (LED) in complete darkness, a subtle, apparent motion is perceived which is called autokinesis. This autokinetic illusion increases with increasing fixation time. Eleven healthy subjects were examined by fMRI while fixating an LED in darkness for 35 s. BOLD signal changes of the first and the second half of the fixation period were compared. While the stimulus was the same for both periods, perception differed in that autokinesis was more pronounced in the second half. This second half of the period was associated with bilateral activations in the motion-sensitive middle occipito-temporal area known as MT/V5. Our finding suggests that area MT/V5 is involved in the mediation of autokinesis. PMID- 15878674 TI - Surface EMG measurements during fMRI at 3T: accurate EMG recordings after artifact correction. AB - In this experiment, we have measured surface EMG of the first dorsal interosseus during predefined submaximal isometric contractions (5, 15, 30, 50, and 70% of maximal force) of the index finger simultaneously with fMRI measurements. Since we have used sparse sampling fMRI (3-s scanning; 2-s non-scanning), we were able to compare the mean amplitude of the undisturbed EMG (non-scanning) intervals with the mean amplitude of the EMG intervals during scanning, after MRI artifact correction. The agreement between the mean amplitudes of the corrected and the undisturbed EMG was excellent and the mean difference between the two amplitudes was not significantly different. Furthermore, there was no significant difference between the corrected and undisturbed amplitude at different force levels. In conclusion, we have shown that it is feasible to record surface EMG during scanning and that, after MRI artifact correction, the EMG recordings can be used to quantify isometric muscle activity, even at very low activation intensities. PMID- 15878675 TI - A whole brain MR spectroscopy study from patients with Alzheimer's disease and mild cognitive impairment. AB - Brain damage in Alzheimer's disease (AD) and mild cognitive impairment (MCI) is widespread with involvement of large portions of the neocortex and the subcortical white matter. A quantitative measure of neuronal damage of the entire brain might be valuable in the context of large-scale, longitudinal studies of these patients. This study investigated the extent of neuroaxonal injury of patients with AD and MCI using a novel unlocalized proton magnetic resonance spectroscopy ((1)H-MRS) technique, which allows quantification of the concentration of N-acetylaspartate from the whole of the brain tissue (WBNAA). Conventional brain MRI and WBNAA were obtained from 28 AD patients, 27 MCI patients and 25 age-matched controls. Normalized brain volume (NBV) was also measured using an automated segmentation technique. WBNAA and NBV showed a significant heterogeneity between groups (P < 0.001). WBNAA concentration was different between controls and MCI patients (P = 0.003), but not between MCI and AD patients (P = 0.33). NBV differed both between controls and MCI patients (P = 0.02) and between MCI and AD patients (P = 0.03). A multivariate regression model retained WBNAA as the best MRI predictor of the Mini Mental State Examination score (P = 0.001). Significant neuronal damage, which is related to the extent of cognitive decline, can be quantified in the whole brain tissue of patients with AD, using a novel (1)H-MRS approach. The demonstration in patients with MCI of MR structural and metabolic findings, intermediate between those of healthy volunteers and those of AD patients, indicates that neuronal damage is already evident and widespread in individuals with MCI before they are clinically demented. PMID- 15878676 TI - 1H-MRS imaging in intractable frontal lobe epilepsies characterized by depth electrode recording. AB - Presurgical evaluation of frontal lobe epilepsy (FLE) remains a challenging issue and frequently requires invasive depth electrode recording. In this study, we aimed at evaluating the potential usefulness of a non-invasive technique such as proton magnetic resonance spectroscopic imaging ((1)H-MRSI) in the presurgical evaluation of FLE and at investigating the potential electrophysiological correlates of the metabolic disturbances as defined by (1)H-MRSI. We compared the distribution of (1)H-MRSI abnormalities with the electrophysiological abnormalities defined by stereo-electroencephalography (SEEG) recording in 12 patients presenting with several subtypes of FLE. We also used 12 control subjects in order to obtain normative (1)H-MRSI data. We used a multilevel (1)H MRSI protocol to better sample the principal regions of the frontal lobe. We also applied a metabolic mapping technique allowing a visual display of metabolic data. A significant decrease of both N-acetyl-aspartate/phosphocreatine-creatine and N-acetyl-aspartate/(choline-compounds + phosphocreatine-creatine) ratios was observed in regions involved in the epileptogenic zone (EZ) and/or the irritative zone (IZ) compared to regions without electrical abnormalities in the same patients (P = 0.044 and P = 0.018, respectively), and also compared to controls (P = 0.004 and P = 0.0001, respectively). No significant differences in metabolic ratios were observed between those regions involved in the EZ and those involved in the IZ only. Our results suggest a link between the relative decrease of N acetyl-aspartate and the EZ as well as the IZ in FLE. Thus, multilevel (1)H-MRSI protocol may add pertinent information during the non-invasive presurgical evaluation of FLE. PMID- 15878677 TI - Neural mechanisms of visual backward masking revealed by high temporal resolution imaging of human brain. AB - Backward masking is one of the potent ways to reveal the neural mechanism of visual awareness in humans. Although previous neuroimaging studies have reported that the visual masking involves the attenuation of hemodynamic signals to the masked stimulus in visual ventral regions such as the fusiform and inferior temporal gyrus, the temporal profiles of this attenuation as a whole neural population is mostly unclear. Here we used magnetoencephalography and investigated the neural response changes in higher visual region induced by backward masking. The combination of our previous random dot blinking method with the sensor-based analysis isolated the neural responses in the higher visual cortex relating to shape perception. The results revealed that, as the visibility of the target stimulus was reduced by the mask following it, the neural response to the target in the ventral regions showed gradual decreases both in its peak amplitude and peak latency. Furthermore, this decrease in the peak amplitudes was significantly correlated with the behavioral accuracy of the target identification, while the peak latency was not. These results indicate that backward masking simultaneously produces two types of neural changes in higher visual regions: attenuation of the populational neural activity itself and temporal interruption of this activity by the subsequent mask response. Especially, our data suggest that the response attenuation in higher visual response is a main cause of the perceptual impairment observed in the backward masking paradigm. PMID- 15878678 TI - Free fatty acid metabolism in the air-breathing African catfish (Clarias gariepinus) during asphyxia. AB - In several waterbreathing fish species, hypoxia induces a decrease in plasma free fatty acid (FFA) levels as opposed to an increase in air-breathing mammals. We hypothesised that this change is coupled to the mode of breathing. Therefore, we followed the metabolic response of cannulated air-breathing African catfish to an 8-h asphyxia period. The hematocrit and hemoglobin increased significantly upon asphyxia. However, no change was observed in the mean cellular hemoglobin concentration, indicating that more erythrocytes were brought into circulation. A continuous increase in plasma lactate concentration during asphyxia showed permanent activation of anaerobic glycolysis, pointing to a persistent oxygen shortage. Plasma glucose levels did not change, but FFA levels decreased significantly upon asphyxia with a concomitant increase in plasma noradrenaline levels. Thus, these results suggest that in the air-breathing African catfish noradrenaline mediated a decrease in plasma FFA levels similar to that in waterbreathing fish species. PMID- 15878679 TI - Antibody responses to individual proteins of SARS coronavirus and their neutralization activities. AB - A novel coronavirus, the severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV), was identified as the causative agent of SARS. The profile of specific antibodies to individual proteins of the virus is critical to the development of vaccine and diagnostic tools. In this study, 13 recombinant proteins associated with four structural proteins (S, E, M and N) and five putative uncharacterized proteins (3a, 3b, 6, 7a and 9b) of the SARS-CoV were prepared and used for screening and monitoring their specific IgG antibodies in SARS patient sera by protein microarray. Antibodies to proteins S, 3a, N and 9b were detected in the sera from convalescent-phase SARS patients, whereas those to proteins E, M, 3b, 6 and 7a were undetected. In the detectable specific antibodies, anti-S and anti-N were dominant and could persist in the sera of SARS patients until week 30. Among the rabbit antisera to recombinant proteins S3, N, 3a and 9b, only anti-S3 serum showed significant neutralizing activity to the SARS-CoV infection in Vero E6 cells. The results suggest (1) that anti-S and anti N antibodies are diagnostic markers and in particular that S3 is immunogenic and therefore is a good candidate as a subunit vaccine antigen; and (2) that, from a virus structure viewpoint, the presence in some human sera of antibodies reacting with two recombinant polypeptides, 3a and 9b, supports the hypothesis that they are synthesized during the virus cycle. PMID- 15878680 TI - Rapid clearance of a recombinant Salmonella vaccine carrier prevents enhanced antigen-specific CD8 T-cell responses after oral boost immunizations. AB - The type III secretion system of Salmonella enterica serovar Typhimurium can be used to target heterologous antigens directly into the cytosol of antigen presenting cells. Our laboratory has previously reported that the single oral immunization of mice with a recombinant Salmonella strain expressing the translocated Yersinia outer protein E fused to the immunodominant antigen p60 from Listeria monocytogenes results in the efficient induction of p60-specific CD8 T cells and confers protection against a lethal wild-type Listeria challenge infection. In the present study, we investigated whether these antigen-specific cytotoxic T lymphocytes induced by the prime immunization contribute to a more rapid clearance of the vaccine carrier after subsequent boost immunizations and whether oral boost immunizations lead to an augmented p60-specific CD8 T-cell response. We found that the ability of recombinant Salmonella strains to colonize the intestine, mesenteric lymph nodes, and spleen was markedly impaired after boost immunizations but that this effect was independent of existing CD8 T cells reactive with p60(217-225). A significant elevation of antigen-specific CD8 T cells could not be detected by enzyme-linked immunospot assay after the second or the third oral immunization, possibly due to the rapid clearance of the bacterial vaccine carrier from lymphatic organs. PMID- 15878681 TI - Internalin-expressing Lactococcus lactis is able to invade small intestine of guinea pigs and deliver DNA into mammalian epithelial cells. AB - The use of the food-grade bacterium Lactococcus lactis as antigen delivery vehicle at the mucosal level is an attractive vaccination strategy intensively explored during the last decade. In this study, we developed L. lactis strains which could be used as a DNA delivery vector to combine both advantages of mucosal delivery and of DNA vaccination. To render lactococci capable of invading epithelial cells, the Listeria monocytogenes inlA gene was cloned and expressed in L. lactis under transcriptional control of the native promoter. Western blot and immunofluorescence assays revealed that recombinant lactococci efficiently displayed the cell wall anchored form of InlA. We demonstrated that this expression promotes internalization of L. lactis inlA+ into the human epithelial cell line Caco-2. Gentamicin assay showed that invasiveness of L. lactis in these cells is approximately 100-fold higher for L. lactis inlA+ than for wild type (wt) L. lactis strains. Moreover, we showed that L. lactis inlA+ is able to enter intestinal cells in vivo, after oral inoculation of guinea pigs. After internalization, L. lactis inlA+ was able to deliver a functional eukaryotic gfp gene into epithelial Caco-2 cells; GFP was detected in 1% of internalized cells. The L. lactis inlA+ strain will be a useful bacterial vector for the development of new live oral DNA vaccines. It also constitutes an interesting new model to study the role of internalin in bacterial localization in the animal host. PMID- 15878682 TI - PorA protein of Campylobacter jejuni is not a cytotoxin mediating inflammatory diarrhoea. AB - Campylobacter jejuni is a major food-borne pathogen and a leading cause of diarrhoea. A cytotoxin is most likely involved in the pathogenesis of inflammatory diarrhoea due to C. jejuni. A 45-kDa outer membrane protein encoded by the porA gene was reported to exhibit cytotoxic activity for cultured mammalian cells in vitro. We cloned and expressed the porA gene in Escherichia coli BL21 codon plus RIL strain using the fusion vector pGEX-4T-1. The fusion protein solubilised in urea in denatured form or solubilised in Empigen BB in native form or their thrombin-cleaved products did not exhibit cytotoxic activity for Chinese hamster ovary (CHO) cells. The urea-solubilised fusion protein did not induce fluid accumulation in the rabbit ileal loop assay. All 76 clinical isolates of Campylobacter spp. tested were positive for porA by PCR, but only 13 isolates were positive for cytotoxin on CHO cells. Both cytotoxin-positive as well as cytotoxin-negative strains expressed PorA as determined by immunoblot analysis. These findings show that the porA gene product of C. jejuni is not a cytotoxin mediating inflammatory diarrhoea. PMID- 15878683 TI - Plasmid DNA vaccination. AB - Plasmid DNA vaccination against tuberculosis is a very powerful and easy method for the induction of strong humoral responses, CD4+ mediated secretion of Th1 cytokines and CD8+ mediated CTL activity in mice. Tuberculosis DNA vaccines have not been assessed so far in humans, and clinical trials with DNA in general have been somewhat disappointing. However, numerous studies have reported on the potent priming capacity of plasmid DNA for Th1 and CD8+ mediated immune responses, which can be boosted subsequently by recombinant protein or recombinant pox-viruses. With respect to tuberculosis, prime/boost regimens with Mycobacterium bovis BCG vaccine are particularly promising and warrant further analysis. PMID- 15878684 TI - Monitoring for human cytomegalovirus infection in solid organ transplant recipients through antigenemia and glycoprotein N (gN) variants: evidence of correlation and potential prognostic value of gN genotypes. AB - Human cytomegalovirus (HCMV) ORF UL73 encodes the envelope glycoprotein gpUL73 gN, which shows seven genotypes (gN-1, gN-2, gN-3a, gN-3b, gN-4a, gN-4b, gN-4c). The goal of this study was to determine retrospectively the distribution of gN variants in solid organ transplant recipients with HCMV infection and to establish an association with parameters important for monitoring post transplantation clinical course during a follow-up of up to 2 years. Peripheral blood leukocytes from 40 solid organ transplant recipients were analysed for pp65 antigen by immunofluorescence and gN genotyped by sequencing or RFLP analysis. A correlation between gN genotypes and antigenemia peak was found, showing a highly significant difference between gN-1 and gN-4b variants (P<0.005). In particular, gN-1 seems to be associated with patients developing low level antigenemia (<50 pp65-positive cells/2 x 10(5) PBLs; PPV = 90%), whereas gN-4b predicts significantly higher values (>50 pp65-positive cells/2 x 10(5) PBLs; PPV = 80%). Furthermore, the onset of positive antigenemia is significantly earlier in patients infected with a gN-4b strain, compared with those infected by a gN-1 variant. Reported data further support a role for gN genotypes in HCMV pathogenesis. gN-1 and gN-4b show a significantly different virulence and could serve as early predictors for the progression of HCMV infection in transplant patients. PMID- 15878685 TI - Introduction. Rational vaccine development against tuberculosis: "Those who don't remember the past are condemned to repeat it". PMID- 15878686 TI - [Preliminary results from the OPNI observatory: long-term follow-up of a cohort of women using the progestagen contraceptive implant Implanon]. AB - OBJECTIVE: Describe the initiation and follow-up of Implanon insertion in current office-based practice in France and estimate the rate and causes of early removals. PATIENTS AND METHODS: A prospective cohort study of 1000 women having been inserted with Implanon by a representative national sample of prescribers (gynaecologists and general practitioners) was designed. The follow-up period was 3 years and the enrolment was planned for 2 years starting July 2002 according to a naturalistic design. RESULTS: The results are related to an intermediate analysis describing the situation of the cohort at the date 31 December 2003. 872 women were enrolled, of whom 784 (89.9%) by gynaecologists and 88 (10.1%) by GPs. Implanon was inserted in 691 (79.2%) and 360 (52.1%) had at least one follow-up visit at this date. The mean follow-up period after insertion was 10.5 months. 38 early removals were recorded (actuarial rate at 16 months of 8.8% [CI 95% 5.0 12.7]), integrating the distribution of follow-up duration and the assumption that women with no follow-up visit the still had device. DISCUSSION AND CONCLUSION: The estimated early removal rate was lower than the result of the meta-analysis of international clinical trials but this figure should be confirmed in the final analysis of the study. PMID- 15878687 TI - [Recording expulsive forces during childbirth using intercostal muscle electromyogram: a pilot study]. AB - OBJECTIVE: The expulsive forces of childbirth can be included among the many potential risk factors implicated in the subsequent development of perineal disorders. The objective of this study was to devise a non-invasive way to measure abdominal pushing that would accurately represent the expulsive forces during childbirth. PATIENTS AND METHODS: By means of intravesical and intrauterine manometry, and electromyography (EMG) of intercostal muscles, we quantified these forces in 21 women during vaginal delivery. RESULTS: A mixed Ancova model showed the integral of intravesical pressure to be significantly associated (P<0.001) with the integral of intercostal muscle electrical activity during the first 6 uterine contractions during the phase of fetal expulsion. DISCUSSION AND CONCLUSION: Electromyography is a non-invasive measurement that can replace intravesical determinations to quantify these forces as it reflects the real intra-abdominal pressure. PMID- 15878688 TI - [Polycythemia vera and pregnancy: difficulties for diagnosis and treatment]. AB - Polycythemia vera is a myeloproliferative syndrome. This clonal disorder involves a pluripotent stem cell capable of differentiating into red blood cells, granulocytes, and platelets. Polycythemia vera is characterized by the overproduction of mature red blood cells in the bone marrow. Myeloid and megakaryocytic elements are also often increased. Polycythemia vera (PV) is rarely associated with pregnancy. About 20 cases have been reported. Prognosis of PV is not influenced by pregnancy. Conversely, pregnancy outcome is poor, due to the occurrence of gestational hypertension, stillbirth and induced prematurity. During pregnancy, clinical management needs to be close including a collaborative approach between obstetricians, hematologists and anesthesists. The risk of poor outcome may be reduced by the association of antiaggregant and anticoagulant therapy. Phlebotomy can be provided in order to maintain hemoglobin level under 42%. PMID- 15878689 TI - Estimation of early postmortem intervals by a multiple regression analysis using rectal temperature and non-temperature based postmortem changes. AB - Five general methods based on rectal temperature and a multiple regression analysis using rectal temperature and non-temperature based postmortem changes were applied to 212 postmortem cases of within 24h postmortem (PM) intervals. Non temperature based postmortem changes of rigidity, hypostasis and corneal turbidity were numerically categorized and used with rectal temperatures as four statistical variables in the multiple regression analysis. The correlation coefficient values between true and calculated postmortem intervals were 0.78 0.82 in the five general methods based on rectal temperature. The multiple regression analysis produced a multiple correlation coefficient value of 0.89 and according to the error ranges of the PM intervals, 72% of the cases were estimated within the error of +/-1.0 h and 92% within +/-5.0 h. Although assessments of non-temperature based PM changes are mostly subjective and have a wide variation, the present study demonstrated a usefulness of non-temperature based PM changes in the estimation of PM intervals. PMID- 15878690 TI - Treatment of acute Sydenham's chorea with methyl-prednisolone pulse-therapy. AB - Serious adverse effects with antichoreic drugs can occur in patients with acute Sydenham's chorea (SC). The response to conventional treatment in severe SC may also be poor. Thus alternative therapeutic strategies have been developed. We report on four patients with severe acute SC (two with chorea paralytica and two with neuroleptic-induced parkinsonism) treated with i.v. methyl-prednisolone pulse-therapy followed by oral prednisone. The mean+/-SEM clinical rating score dropped from 63.7+/-10.2 to 33.5+/-10.6 in the first month post-treatment but in a lesser degree thereafter. Only one patient developed weight gain and 'moon facies'. This case series suggests that immune suppression with corticosteroids may be an effective and safe treatment in complicated and severe acute SC. PMID- 15878691 TI - Acetaminophen decreases intracellular glutathione levels and modulates cytokine production in human alveolar macrophages and type II pneumocytes in vitro. AB - Recent epidemiological observations suggest that acetaminophen (paracetamol) may contribute to asthma morbidity. Impaired endogenous antioxidant defences may have a role in the pathogenesis of a number of inflammatory pulmonary diseases, including asthma. We studied the effect of acetaminophen on the intracellular level of reduced glutathione (GSH) with and without inhibitors of cytochrome P450 or prostaglandin H synthetase, and TNF-alpha, IL-6 and IL-8 protein production in human alveolar macrophages and type II pneumocytes in vitro. Following a 20 h incubation with acetaminophen, cytotoxicity was apparent from > or = 5 and > or = 10 mM in macrophages and type II pneumocytes, respectively. A time- and concentration-dependent decrease of intracellular GSH occurred after acetaminophen (0.05-1 mM) exposure (1-4 h) in pulmonary macrophages (up to 53%) and type II pneumocytes (up to 34%). Diethyldithiocarbamic acid, potassium ethyl xanthate, and indomethacin decreased significantly acetaminophen-induced GSH depletion in the two cell types tested, suggesting the involvement of cytochrome P450 (mainly CYP2E1) and/or prostaglandin H synthetase. In macrophages, acetaminophen decreased the secretion of TNF-alpha (at 4 and 24 h, concentration related) and IL-6 (at 24 h, at 0.1 mM), and did not affect significantly IL-8 production. These in vitro observations demonstrate that clinically relevant concentrations of acetaminophen decreased: (i) intracellular GSH in human pulmonary macrophages and type II pneumocytes and (ii) the secretion of TNF-alpha and possibly IL-6 by human pulmonary macrophages. These findings provide experimental plausibility to the challenging observations that frequent use of APAP may be a risk factor for asthma morbidity. PMID- 15878692 TI - Deficiency of PPARalpha disturbs the response of lipogenic flux and of lipogenic and cholesterogenic gene expression to dietary cholesterol in mouse white adipose tissue. AB - PPARalpha-deficiency in mice fed a high-carbohydrate, low-cholesterol diet was associated with a decreased weight of epididymal adipose tissue and an increased concentration of adipose tissue cholesterol. Consumption of a high (2% w/w) cholesterol diet resulted in a further increase in the concentration of cholesterol and a further decrease in epididymal fat pad weight in PPARalpha-null mice, but had no effect in the wild-type. These reductions in fat pad weight were associated with an increase in hepatic triacylglycerol content, indicating that both PPARalpha-deficiency and cholesterol altered the distribution of triacylglycerol in the body. Adipose tissue de novo lipogenesis was increased in PPARalpha-null mice and was further enhanced when they were fed a cholesterol rich diet; no such effect was observed in the wild-type mice. The increased lipogenesis in the chow-fed PPARalpha-null mice was accompanied paradoxically by lower mRNA expression of SREBP-1c and its target genes, acetyl-CoA carboxylase and fatty acid synthase. Consumption of a high-cholesterol diet increased the mRNA expression of these genes in the PPARalpha-deficient mice but not in the wild-type. De novo cholesterol synthesis was not detectable in the adipose tissue of either genotype despite a relatively high expression of the mRNA's encoding SREBP-2 and 3-hydroxy-3-methylglutaryl Coenzyme A reductase. The mRNA expression of these genes and of the LDL-receptor in adipose tissue of the PPARalpha deficient mice was lower than that of the wild-type and was not downregulated by cholesterol feeding. The results suggest that PPARalpha plays a role in adipose tissue cholesterol and triacylglycerol homeostasis and prevents cholesterol mediated changes in de novo lipogenesis. PMID- 15878693 TI - Presence and potential signaling function of N-acylethanolamines and their phospholipid precursors in the yeast Saccharomyces cerevisiae. AB - N-acylethanolamines (NAEs) and N-acylphosphatidylethanolamines (NAPEs) are trace constituents of vertebrate cells and tissues and much is known about their metabolism and possible function in animals. Here we report for the first time the identification and quantification of NAEs and NAPEs in several strains of the yeast Saccharomyces cerevisiae. Gas chromatography-mass spectrometry of appropriate derivatives revealed 16:0, 16:1, 18:0 and 18:1 N-acyl groups in both NAE and NAPE whose levels, in wild-type cells, were 50 to 90 and 85 to 750 pmol/micromol lipid P, respectively (depending on the phase of growth). NAPE levels were reduced by 45 to 60% in a strain lacking three type B phospholipases, suggesting their involvement in NAPE synthesis by their known transacylation activity. A yeast strain lacking the YPL103c gene, which codes for a protein with 50.3% homology to human NAPE-specific phospholipase D, exhibited a 60% reduction in NAE, compared to wild-type controls. The exposure of various yeast strains to peroxidative stress, by incubation in media containing 0.6 mM H(2)O(2), resulted in substantial increases in NAE. Because yeast cells lack polyunsaturated fatty acids, they offer a useful system for the study of NAE generation and its potential signaling and cytoprotective effects in the absence of polyunsaturated ("endocannabinoid") congeners. PMID- 15878694 TI - 10 challenges in supervision of doctoral students. PMID- 15878695 TI - Upregulation of estrogen receptor alpha and vitellogenin in eelpout (Zoarces viviparus) by waterborne exposure to 4-tert-octylphenol and 17beta-estradiol. AB - The mechanisms of action of an estrogenic chemical have been examined in a viviparous fish the eelpout (Zoarces viviparus), by identification of an upregulated estrogenic pathway--the induction of hepatic estrogen receptor mRNA, hepatic estrogen binding activity and plasma vitellogenin. A relative quantitative RT-PCR assay has been established to measure hepatic estrogen receptor alpha (ER) mRNA levels in eelpout. Assay conditions were optimised using control and induced samples to ascertain its applicability in the actual working range of ER mRNA concentrations. beta-Actin was co-amplified and used as an internal standard. Time-course effects of water exposure to 0.5 microg/L 17beta estradiol (E(2)) and 25 microg/L of the xeno-estrogen 4-tert-octylphenol (4-tert OP) on ER mRNA levels in the male eelpout was examined. After 48 h of exposure, ER transcripts were induced 15-fold and 6-fold in the E(2)- and OP-treated fish, respectively. This difference, however, was not apparent after 1 week of exposure, when similar high levels of ER mRNA were present in both groups (20 fold induction). This indicates that the estrogenic capacity of 4-tert-OP increases with exposure time. The effect of treatment was also evaluated by examining the induction of specific E(2) binding capacity in hepatic cytosolic extracts and by measuring vitellogenin in plasma. Both parameters were also induced by the treatments, but later in the time course. The measurement of ER mRNA by the RT-PCR assay showed to be the most sensitive method for the detection of estrogenic responses in eelpout. PMID- 15878696 TI - Role of mitochondrial DNA in toxic responses to oxidative stress. AB - Mitochondria are at the crossroads of several crucial cellular activities including: adenosine triphosphate (ATP) generation via oxidative phosphorylation; the biosynthesis of heme, pyrimidines and steroids; calcium and iron homeostasis and programmed cell death (apoptosis). Mitochondria also produce considerable quantities of superoxide and hydrogen peroxide (H2O2) that in conjunction with its large iron stores can lead to a witch's brew of reactive intermediates capable of damaging macromolecules. Mitochondrial DNA (mtDNA) represents a critical target for such oxidative damage. Once damaged, mtDNA can amplify oxidative stress by decreased expression of critical proteins important for electron transport leading to a vicious cycle of reactive oxygen species (ROS) and organellar dysregulation that eventually trigger apoptosis. Oxidative stress is associated with many human disorders including: cancer, cardiovascular disease, diabetes mellitus, liver disease and neurodegenerative disease. This article reviews the evidence that oxidative damage to mtDNA can culminate in cell death and thus represents an important target for therapeutic intervention in a number of human diseases. PMID- 15878697 TI - Physiological and pathophysiological down-regulation of cough. AB - Recent clinical studies have emphasized the up-regulation (sensitization) of cough in pathological conditions of the airways. However there are also many situations where voluntary and reflex cough can be down-regulated. These include: (1) chemical stimulation of breathing by hypercapnia or hypoxia or both, establishing that cough sensitivity can be inversely related to drive to breathing; (2) voluntary inhibition of cough, probably similar in mechanism to the depression of cough that can be induced by hypnosis and other branches of alternative medicine; (3) the placebo effect of many antitussive treatments; (4) sleep; (5) general anaesthesia; (6) central nervous disorders such as coma, stroke, Parkinson's disease and several other conditions where the defect in the protective reflexes may lead to aspiration pneumonia; (7) increased activity in various afferent inputs from viscera in the thorax and abdomen; (8) a number of bronchopulmonary clinical disorders. The list is long, but regrettably the nervous mechanisms of these down-regulations have been little studied. In addition there are a number of situations, such as exercise, coitus, talking and singing which, while important to coughing humans, have been not investigated in relation to cough. Most of the studies have been with experimental animals, and their extension to human research is desirable. In view of the importance of cough and other defensive reflexes in maintaining human well-being, far more research is needed. The field is wide-open. PMID- 15878698 TI - Crystallization and preliminary X-ray diffraction analysis of importin-alpha complexed with NLS peptidomimetics. AB - Importin-alpha is the nuclear import receptor that recognizes cargo proteins with nuclear localization sequences (NLSs). The study of NLS peptidomimetics can provide a better understanding of the requirements for the molecular recognition of cargo proteins by importin-alpha, and potentially engender a large number of applications in medicine. Importin-alpha was crystallized with a set of six NLS peptidomimetics, and X-ray diffraction data were collected in the range 2.1-2.5 A resolution. Preliminary electron density calculations show that the ligands are present in the crystals. PMID- 15878699 TI - NMR and mass spectrometry studies of putative interactions of cell cycle proteins pRb and CDK6 with cell differentiation proteins MyoD and ID-2. AB - Cell growth and differentiation require precise coordination of cell cycle and differentiation proteins. This can be achieved by direct interactions between proteins, by indirect interaction in multiprotein complexes, or by modulation of gene expression levels of partner proteins. Contradictory data abound in the literature regarding the binding between some central cell cycle proteins, pRb, and CDK6, with myogenic differentiation promoting, MyoD, and inhibiting, Id-2, factors. We have tested these interactions using pure proteins and in vitro biophysical and biochemical methods, which included mass spectrometry, nuclear magnetic resonance (NMR), the affinity chromatography pull-down assays, and gel filtration chromatography. Using this multimethod approach, we were able to document interactions between pRb and HPV-E7, pRb and SV40 large T antigen, CDK6 and p19, and MyoD and DNA. Using the same methods, we could unambiguously show that there is no direct protein-protein interaction in vitro between the small pocket domain of pRb and the bHLH domain of MyoD, the small pocket domain of pRb and Id-2, and CDK6 and a 15-amino-acid peptide from the C-terminal domain of MyoD. Indirect interactions, through additional binding partners in multiprotein complexes or modulation of gene expression levels of these proteins, are therefore their probable mode of action. PMID- 15878700 TI - Skeletal dysplasias. AB - This review is intended to help the neonatologist who is asked to see a baby or speak to parents who are expecting a baby with signs of a generalised disturbance of bone growth and/or modelling. In this review, we will: define a skeletal dysplasia; discuss the presenting features of a skeletal dysplasia in pregnancy and the newborn period; suggest a clinical approach to find the correct diagnosis; discuss the management of the neonate with a skeletal dysplasia; summarise the clinical features of the most common dysplasias; outline some pitfalls and difficulties in counselling the parents of the baby; and give information on further sources of information about skeletal dysplasias. PMID- 15878701 TI - Dental phenotype of the col1a2(oim) mutation: DI is present in both homozygotes and heterozygotes. AB - Dentinogenesis imperfecta (DI) is a common but variable feature of osteogenesis imperfecta (OI). The Col1a2(oim) mutation (oim) is a well-studied mouse model of chain deficiency OI. Heterozygous oim/+ mice have subtle skeletal fragility, while homozygous oim/oim mice have marked skeletal fragility. To further define the consequences of oim mutation, we examined teeth by light and scanning electron microscopy (SEM). The dental phenotype in Col1a2(oim) (oim) mice is more severe in incisors than in molars and includes changes in pulp chamber size, tooth shape, and dentin ultrastructure. Teeth in oim/oim animals are clinically fragile, while oim/+ teeth are grossly normal. Incisor pulp chamber areas (in mum(2)) are: upper +/+ = 358 +/- 75, lower +/+ = 671 +/- 162, upper oim/+ = 161 +/- 54, lower oim/+ = 156 +/- 19, upper oim/oim = 6900 +/- 1040, and lower oim/oim = 66 +/- 62 (P < 10(-5)). Incisor non-pulp chamber cross-sectional areas (in mum(2)), reflecting dentin areas, are: upper +/+ = 39,000 +/- 1670, lower +/+ = 35,600 +/- 1980, upper oim/+ = 47,500 +/- 2510, lower oim/+ = 26,000 +/- 1830, upper oim/oim = 29,800 + 315, and lower oim/oim = 36,800 +/- 3450 (P < 10(-5)). Ultrastructural abnormalities are more pronounced in incisors than in molars and depend on dosage of the mutant allele. These include reduction in the number and regularity of spacing of the dentinal tubules, lesser mineralization, and blurring of the boundary between peritubular and intertubular dentin. Our findings demonstrate that both oim/oim and oim/+ mice suffer from DI. The more severe incisor phenotype may reflect incisors' continuous growth. PMID- 15878702 TI - The morphological association between microcracks and osteocyte lacunae in human cortical bone. AB - We studied the spatial relationship between the osteocyte lacunar-canalicular network and microdamage accumulation in bone matrix. Rib sections from 9 white women aged 50-60 were stained with basic fuchsin and examined using bright-field and fluorescence microscopy. The results showed that the numerical and length density of cracks were 5-fold higher in interstitial bone than in osteons (P<0.001). Osteocyte lacunar density was 17% lower in interstitial bone than in osteonal bone (P<0.001). In addition, the osteocyte lacunae in interstitial bone were significantly fewer (by 16%) in the area adjacent to microdamage as compared with the area remote from microdamage (P<0.001). The proportion of fields with lacunar density less than 728/mm2, the cut-off point calculated from ROC analysis, was 30% in osteonal bone, 55% in interstitial bone remote from microcracks and 83% adjacent to microcracks. The mean values of lacunar density in these bones were 10%, 22% and 27% lower than the cut-off point, respectively. The likelihood of microdamage was 3.8 times higher in bone with osteocyte lacunar density <728/mm2. About 73% of the crack profiles were spatially associated, at least partly, with bone fragments in which osteocyte lacunae were absent. We conclude that microdamage and osteocyte deficiency occur in the same bone regions; there is likely a causal relationship between them but we are unable to say which comes first. PMID- 15878703 TI - A statistical profile of road accidents during cross-flow turns. AB - In-depth studies of behavioral factors in road accidents using conventional methods are often inconclusive and costly. In a series of studies exploring alternative approaches, 200 cross-flow junction road accidents were sampled from the files of Nottinghamshire Constabulary, England, coded for computer analysis using a specially devised 'Traffic Related Action Analysis Language', and then examined using different computational and statistical techniques. For comparison, the same analyses were also carried out on 100 descriptions of safe turns, and 100 descriptions of hypothetical accidents provided by experienced drivers. The present study used statistical methods to explore the database of cases. The youngest and oldest groups of drivers were found to be over represented in the junction accidents, and were the least likely to stop before turning. The young drivers had particular problems turning onto major roads. Women were more likely than men to stop before turning; they tended to have their collisions with other women; and they were under-represented as drivers of the non-turning vehicle. In hypothetical accidents, informants tended to blame the younger driver, increasingly so for male informants as they got older. Female informants tended to blame male drivers. PMID- 15878704 TI - Protocatechuic aldehyde suppresses TNF-alpha-induced ICAM-1 and VCAM-1 expression in human umbilical vein endothelial cells. AB - Adhesion molecules, which play a crucial role in the development of atherogenesis, are produced by endothelial cells following stimulation with various inflammatory cytokines. The current studies examined the effect of a potent water-soluble antioxidant, protocatechuic aldehyde (derived from the Chinese herb, Salvia miltiorrhiza), on the expression of adhesion molecules in human umbilical vein endothelial cells (HUVECs) stimulated with tumor necrosis factor-alpha (TNF-alpha). Protocatechuic aldehyde appeared to specifically downregulate the TNF-alpha-induced cell surface expression of vascular adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) on HUVECs as well as the release of soluble VCAM-1and ICAM-1 from HUVECs in a dose-response manner at pharmacologically relevant concentrations (0.15-1.35 mM). We also observed a dose-dependent lowering of mRNA expression of VCAM-1 and ICAM-1 in the presence of protocatechuic aldehyde. Furthermore, protocatechuic aldehyde (0.15, 0.45, and 1.35 mM) notably inhibited TNF-alpha-induced upregulation of U937 cell adhesion to HUVECs to 83.7%, 60.9%, and 40.8%, respectively. A gel shift assay further showed that protocatechuic aldehyde inhibited the TNF-alpha-activated NF kappaB and AP-1 DNA binding activities in a dose-dependent manner. Collectively, these results indicate that protocatechuic aldehyde inhibits TNF-alpha-stimulated VCAM-1 and ICAM-1expression in HUVECs through a mechanism that involves NF-kappaB and AP-1. PMID- 15878705 TI - cGMP-phosphodiesterase antagonists inhibit Ca2+-influx in Dictyostelium discoideum and bovine cyclic-nucleotide-gated-channel. AB - We used antagonists of cGMP-phosphodiesterases to examine the role of cGMP for light-scattering oscillations and cAMP-induced Ca(2+)-influx in Dictyostelium discoideum, however, SCH 51866 (cis-5,6a,7,8,9,9a-hexahydro-2-[4 (trifluoromethyl)phenylmethyl]-5-methyl-cyclopent[4,5]imidazo[2,1-b]purin-4(3H) one) and sildenafil citrate (1-[[3-(6,7-dihydro-1-methyl-7-oxo-3-propyl-1-H pyrazolo[4,3-d]pyrimidin-5-yl)-4-ethoxyphenyl]sulfonyl]-4-methylpiperazine citrate) were poor inhibitors of cGMP-hydrolysis. Instead, SCH 51866 (IC(50) = 16 microM) and sildenafil, blocked chemoattractant (cAMP)-induced Ca(2+)-influx as determined with a Ca(2+)-specific electrode. SCH 51866 (150 microM) affected neither spontaneous cGMP transients during light-scattering-oscillations nor cAMP mediated K(+)-efflux. SCH 51866 and sildenafil are competitive inhibitors of cGMP phosphodiesterases. However, the activity of cGMP-dependent protein kinase Ialpha (PKGIalpha) was not altered by SCH 51866 (150 microM). By contrast, patch-clamp measurements of bovine cone cGMP-gated-channels (cyclic-nucleotide-gated-channel, CNGA3), stably expressed in human embryonic kidney cells, HEK 293 cells, revealed reversible, competitive and dose-dependent inhibition of sodium currents by SCH 51866 (IC(50) = 25 microM) and sildenafil, but not by another inhibitor of cGMP phosphodiesterases, UK 114,542. The possibility that D. discoideum cells also express a cGMP-regulated channel is supported by our finding that LY 83583 (6 (phenylamino)-5,8-quinolinedione) (35 microM), known to inhibit cyclic-nucleotide gated-channels as well as guanylyl-cyclases, reduced cAMP-induced Ca(2+)-influx in D. discoideum, but did not affect cAMP-induced cGMP accumulation. Utilizing a PDED null strain that exhibits a prolonged and elevated cGMP transient following receptor activation, we found that the inhibition of Ca(2+)-influx by SCH 51866 in the wildtype was absent in the mutant. Our results show that SCH 51866 and sildenafil are antagonists of a Ca(2+)-permeable channel (CNGA3) and that both compete with cGMP for a regulatory site of Ca(2+)-influx in D. discoideum. PMID- 15878706 TI - Gene expression of energy and protein metabolism in hearts of hypertensive nitric oxide- or GSH-depleted mice. AB - Hypertension demands cardiac synthetic and metabolic adaptations to increased afterload. We studied gene expression in two models of mild hypertension without overt left ventricular hypertrophy using the NO synthase inhibitor nitro-L arginine (L-NNA) and the glutathione depletor buthionine-S,R-sulfoximine (BSO). Mice were administered L-NNA, BSO, or water for 8 weeks. RNA of left ventricles was pooled per group, reverse transcribed, Cy3 and Cy5 labeled, and hybridized to cDNA microarrays. Normalized log(2) Cy3/Cy5 ratios of > or =0.7 or < or =-0.7 were considered significant. L-NNA and BSO both caused hypertension. Gene expression was regulated in cytoskeletal components in both models, protein synthesis in L-NNA-treated mice, and energy metabolism in BSO-treated mice. Energy metabolism genes shared several common transcription factor-binding sites such as Coup-Tf2, of which gene expression was increased in BSO-treated mice, and COMP-1. Characterization of the left ventricular adaptations as assessed with gene expression profiles reveals differential expression in energy and protein metabolism related to the pathogenetic background of the hypertension. PMID- 15878707 TI - Ligand binding and functional properties of human angiotensin AT1 receptors in transiently and stably expressed CHO-K1 cells. AB - Chinese Hamster Ovary Cells (CHO-K1) were transiently and stably transfected to express the human angiotensin AT(1) receptor. Cell surface receptor expression was maximal 2 days after transient transfection. Their pharmacological and signalling properties differed from stably expressed receptors. Receptor reserve was significant in the transient cells but not in stable cells, explaining the higher potency of angiotensin II and the lower degree of insurmountable inhibition by candesartan in the transient cells. [Sar(1)Ile(8)]angiotensin II (sarile) is a potent angiotensin AT(1) receptor antagonist for the stable cells but is a partial agonist, producing 19% of the maximal response by angiotensin II, in transient cells. Internalization of [(3)H]angiotensin II and [(125)I]sarile (i.e., acid-resistant binding) was more pronounced in stable cells. CHO-K1 cells were also transiently transfected with the enhanced green fluorescence-AT(1) receptor gene. Confocal microscopy revealed rapid internalization induced by angiotensin II and sarile but not by candesartan. The above disparities may result from differences in receptor maturation and/or cellular surrounding. PMID- 15878708 TI - Iron complexing activity of mangiferin, a naturally occurring glucosylxanthone, inhibits mitochondrial lipid peroxidation induced by Fe2+-citrate. AB - Mangiferin, a naturally occurring glucosylxanthone, has been described as having antidiabetic, antiproliferative, immunomodulatory and antioxidant activities. In this study we report for the first time the iron-complexing ability of mangiferin as a primary mechanism for protection of rat liver mitochondria against Fe(2+) citrate induced lipid peroxidation. Thiobarbituric acid reactive substances and antimycin A-insensitive oxygen consumption were used as quantitative measures of lipid peroxidation. Mangiferin at 10 microM induced near-full protection against 50 microM Fe(2+)-citrate-induced mitochondrial swelling and loss of mitochondrial transmembrane potential (DeltaPsi). The IC(50) value for mangiferin protection against Fe(2+)-citrate-induced mitochondrial thiobarbituric acid reactive substance formation (9.02+/-1.12 microM) was around 10 times lower than that for tert-butylhydroperoxide mitochondrial induction of thiobarbituric acid reactive substance formation. The xanthone derivative also inhibited the iron citrate induction of mitochondrial antimycin A-insensitive oxygen consumption, stimulated oxygen consumption due to Fe(2+) autoxidation and prevented Fe(3+) ascorbate reduction. Absorption spectra of mangiferin-Fe(2+)/Fe(3+) complexes also suggest the formation of a transient charge transfer complex between Fe(2+) and mangiferin, accelerating Fe(2+) oxidation and the formation of a more stable Fe(3+)-mangiferin complex unable to participate in Fenton-type reaction and lipid peroxidation propagation phase. In conclusion, these results show that in vitro antioxidant activity of mangiferin is related to its iron-chelating properties and not merely due to the scavenging activity of free radicals. These results are of pharmacological relevance since mangiferin and its naturally contained extracts could be potential candidates for chelation therapy in diseases related to abnormal intracellular iron distribution or iron overload. PMID- 15878709 TI - Cloning and functional characterization of dog transient receptor potential vanilloid receptor-1 (TRPV1). AB - Transient receptor potential vanilloid receptor-1 (TRPV1) is a sensory neuron specific cation channel capable of integrating various noxious chemical and physical stimuli. The dog orthologue of TRPV1 was cloned using cDNA from nodose ganglia and heterologously expressed in HEK293(OFF) cells. At the amino acid level, dTRPV1 displays 85-89% sequence identity to other TRPV1 orthologues. Molecular pharmacological characterization of HEK293(OFF) cells expressing TRPV1 was assessed using a fluorescence imaging plate reader (FLIPR)-based calcium imaging assay. Dog TRPV1 was activated by various known TRPV1 agonists in a concentration-dependent manner: Ag23 = resiniferatoxin > olvanil approximately arvanil > capsaicin > phorbol 12-phenylacetate 13-acetate 20-homovanillate (PPAHV) > N-oleoyldopamine (OLDA). In addition, select TRPV1 antagonists (capsazepine, I-resiniferatoxin and N-(-4-tertiarybutylphenyl)-4-(3-cholorpyridin 2-yl)tetrahydropyrazine-1(2H)-carbox-amide (BCTC)) were able to block the response of dTRPV1 to capsaicin. Furthermore, the dog TRPV1 lacked a conserved protein kinase A (PKA) phosphorylation site (117) found in other cloned orthologues, which may have physiological consequences on dog TRPV1 function. Taken together, these data constitute the first study of the cloning, expression and pharmacological characterization of dog TRPV1. PMID- 15878710 TI - Residues stabilizing the heme moiety of the nitric oxide sensor soluble guanylate cyclase. AB - Soluble guanylate cyclase, a heterodimer consisting of an alpha- and a heme containing beta-subunit, is the major receptor for the biological messenger nitric oxide (NO) and is involved in various signal transduction pathways. The heme moiety of the enzyme is bound between the axial heme ligand histidine(105) and the recently identified counterparts of the heme propionic acids, tyrosine(135) and arginine(139). The latter residues together with an invariant serine(137) form the unique heme binding motif Y-x-S-x-R. In this work, we show that replacement of the serine(137) with alanine destabilizes the binding of the heme moiety and impairs NO-mediated soluble guanylate cyclase activation. PMID- 15878711 TI - Systemically administered glucosamine-kynurenic acid, but not pure kynurenic acid, is effective in decreasing the evoked activity in area CA1 of the rat hippocampus. AB - The metabolism of tryptophan along the kynurenine pathway yields several neuroactive intermediates, including kynurenic acid, which is one of the few known endogenous N-methyl-d-aspartate receptor inhibitors; in parallel with this, it is an alpha7 nicotinic acetylcholinergic receptor antagonist. On the basis of these properties, kynurenic acid might therefore come into consideration as a therapeutic agent in certain neurobiological disorders. However, the use of kynurenic acid as a neuroprotective agent is practically excluded because kynurenic acid hardly crosses the blood-brain barrier. We recently synthetized a new compound, glucosamine-kynurenic acid, which is presumed to cross the blood brain barrier more easily. In this study, the effects of systemically administered kynurenic acid and glucosamine-kynurenic acid on CA3 stimulation evoked population spike activity in region CA1 of the rat hippocampus were compared. The effect of kynurenic acid or glucosamine-kynurenic acid was augmented by probenecid (200 mg/kg), which inhibits kynurenic acid excretion from the cerebrospinal fluid. The results showed that, while kynurenic acid administered i.p. or i.v. in doses of 17, 34, 68 or 136 micromol/kg did not cause any observable change in the animals, 136 micromol/kg glucosamine-kynurenic acid (either i.p. or i.v.) resulted in the sudden death of all the animals. The dose of 68 micromol/kg i.v., but not i.p., resulted in a sudden stoppage of breath, but the animals could be reanimated. As small a dose of glucosamine-kynurenic acid as 17 micromol/kg i.p. resulted in a reduction in population spike amplitudes; this effect was further augmented by probenecid, whereas neither 17 micromol/kg nor higher doses of pure kynurenic acid had a similar effect. The results presented here suggest that glucosamine-kynurenic acid passes the blood brain barrier much more readily than does kynurenic acid. PMID- 15878712 TI - Peripheral and spinal mechanisms of antinociceptive action of lumiracoxib. AB - The possible participation of the nitric oxide (NO)-cyclic GMP-K(+) channel pathway, serotonergic or opioidergic system on lumiracoxib-induced local or intrathecal antinociception was assessed in the formalin test. Local or intrathecal administration of lumiracoxib dose-dependently produced antinociception in the second phase of the test. Moreover, local or intrathecal pretreatment with N(G)-L-nitro-arginine methyl ester (L-NAME, NO synthesis inhibitor), 1H-(1,2,4)-oxadiazolo(4,2-a)quinoxalin-1-one (ODQ, guanylyl cyclase inhibitor), glibenclamide (ATP-sensitive K(+) channel blocker), charybdotoxin and apamin (large- and small-conductance Ca(2+)-activated-K(+) channel blockers, respectively) or margatoxin (voltage-dependent K(+) channel blocker), but not N(G)-D-nitro-arginine methyl ester (D-NAME) or vehicle, significantly prevented lumiracoxib-induced antinociception. The intrathecal injection of methiothepin (serotonin receptor antagonist) reduced lumiracoxib-induced intrathecal antinociception. Local peripheral or intrathecal naloxone did not modify either local or intrathecal lumiracoxib-induced antinociception. Results suggest that lumiracoxib activates the NO-cyclic GMP-K(+) channels to produce local and intrathecal antinociception. Data also suggest that lumiracoxib activates the intrathecal serotonergic system, but not opioid receptors either at peripheral or spinal sites. PMID- 15878713 TI - Enhanced reactivity to vasopressin in rat basilar arteries during vasospasm after subarachnoid hemorrhage. AB - Subarachnoid hemorrhage increases the plasma level of vasopressin, a well-known vasoconstrictor. We examined the sensitivity to vasopressin in rat basilar artery after subarachnoid hemorrhage using a rat subarachnoid hemorrhage model. Vasospasm was observed 1-2 days after subarachnoid hemorrhage induction, and the contractile response to vasopressin in rat basilar arteries was assessed. The concentration-response curve for vasopressin in subarachnoid hemorrhage (1 day) rats shifted leftward compared with that of control rats. The concentration response curve for vasopressin V(1) receptor agonist also shifted leftward and upward compared with that of control rats. The concentration-response curve for vasopressin was inhibited not by vasopressin V(2) receptor antagonist but by vasopressin V(1) receptor antagonist. Thus, it was demonstrated that the vasoconstricting effect of vasopressin was significantly enhanced in the vasospasm phase after subarachnoid hemorrhage. PMID- 15878714 TI - Relaxations to oestrogen receptor subtype selective agonists in rat and mouse arteries. AB - It has been recently reported that the oestrogen receptor alpha agonist PPT (4,4',4"-(4-propyl-[1H]-pyrazole-1,3,5-triyl) tris-phenol) is more potent than the oestrogen receptor beta agonist DPN (2,3-bis(4-hydroxyphenyl)-propionitrile) at producing relaxations in rat mesenteric artery. We have investigated the relaxant actions of PPT and DPN in rat and mouse aorta and mesenteric artery. In rat aortic rings contracted with KCl (40 mM), the oestrogen receptor beta agonist DPN produced significantly greater relaxations than the oestrogen receptor alpha agonist PPT. In wild-type (WT) mouse aorta, the same result was found, but in WT mouse mesenteric artery, as in rat mesenteric artery, DPN was significantly less potent than PPT in females but had similar potency to PPT in males. Relaxations to DPN also occurred in aorta from nitric oxide synthase-3-knockout (NOS-3-KO) mice, and in denuded aorta from both mouse and rat. Hence, in the mouse mesenteric artery, as in the rat mesenteric artery, PPT is at least as potent as DPN at producing relaxations; however, DPN was much more potent than PPT in the rat and mouse aorta. Effects of oestrogen receptor subtype selective agonists are tissue dependent. In addition, actions are largely endothelium-independent. PMID- 15878715 TI - Evidence for two atypical conformations of beta-adrenoceptors and their interaction with Gi proteins. AB - In this study, we investigated whether the responses of right atria from sinoaortic denervated rats to CGP12177 (4(3-t-butylamino-2-hydroxypropoxy benzidimidazole-2 one, hydrochloride)), isoprenaline and norepinephrine desensitized in parallel and whether CGP12177 interacted with distinct conformations of beta-adrenoceptors. Right atria from rats 48 h after sinoaortic denervation were subsensitive to isoprenaline, norepinephrine and CGP12177. One week after sinoaortic denervation, the sensitivity to CGP12177 had recovered whereas the responses to isoprenaline and norepinephrine were still subsensitive, suggesting that the binding sites for these molecules showed independent behavior. In atria from 48 h sinoaortic-denervated rats, propranolol or 3 microM CGP20712A (2-hydroxy-5(2-((2-hydroxy-3-(4-((methyl-4-trifluormethyl)1H imidazole 2-yl)-phenoxypropyl) amino) ethoxy)-benzamide monomethane sulphonate)) blocked the responses to 10 nM-1 microM CGP12177 and steepened the curves. The concentration-response curves to CGP12177 in the presence of ICI118,551 (erythro DL-1(-methylindan-4-yloxy)-3-isopropylamino-butan-2-ol) were biphasic, suggesting that CGP12177 interacted with at least two conformations of beta-adrenoceptors that showed negative cooperativism, one acting through beta(2)-adrenoceptor-Gi and the other via beta(1)-adrenoceptor-Gs. This hypothesis was confirmed in right atria from sinoaortic-denervated rats treated with pertussis toxin. PMID- 15878716 TI - Formation of releasable NO stores by S-nitrosoglutathione in arteries exhibiting tolerance to glyceryl-trinitrate. AB - S-Nitrosating nitric oxide (NO) donors like S-nitrosoglutathione (GSNO) induce a persistent inhibition of vascular tone, through the formation of releasable NO stores. In this study, we investigate whether GSNO also induces NO stores-related effects in vessels exhibiting tolerance to glyceryl-trinitrate. Rat aortic rings treated with glyceryl-trinitrate (100 microM for 1 h) exhibited increased level of superoxide and a decrease in NO elevation and relaxation induced by subsequent addition of glyceryl-trinitrate. In glyceryl-trinitrate-treated rings as in controls, pre-exposure to GSNO (1 microM for 30 min) induced a persistent hyporesponsiveness to noradrenaline and a relaxant response to N-acetylcysteine (a low molecular weight thiol which can displace NO from NO stores), both of which being inhibited by guanylyl-cyclase or cyclic GMP-dependent protein kinase inhibitors. These data indicate that GSNO can promote the formation of releasable NO stores in arteries exhibiting increased superoxide level and tolerance to glyceryl-trinitrate. Formation of releasable NO stores is of potential interest to restore the protective effect of NO in organic nitrate-tolerant blood vessels. PMID- 15878717 TI - The role of substance P and bradykinin in the cough reflex and bronchoconstriction in guinea-pigs. AB - In this study we investigated the ability of aerosolized substance P to induce either cough or bronchoconstriction in guinea-pigs. We have also examined whether pre-treatment, by the inhaled route, of animals with a combination of the neutral endopeptidase inhibitor, phosphoramidon (10(-3) M), and the diaminopeptidase IV inhibitor, diprotin A (10(-3) M), enhances the airway response to substance P. Moreover, we also assessed whether aerosol pre-treatment of guinea-pigs with either substance P or bradykinin, at 10(-4) M, affects the citric acid-induced cough and/or bronchoconstriction. Challenge of guinea-pigs with substance P only at 10(-3) M resulted in significant bronchconstriction but only a weak and variable cough response (1.1+/-0.6; P>0.05). Pre-treatment of guinea-pigs with both phosphoramidon and diprotin A resulted in a small non-significant increase in the cough response (2.8+/-0.9 vs. 1.1+/-0.6; P>0.05) but significantly enhanced substance P-induced bronchoconstriction (P<0.05). Moreover, exposure of guinea-pigs to substance P (10(-4) M) prior to citric acid challenge (0.6 M) resulted in a significant (P<0.05) enhancement of the citric acid-induced bronchoconstriction but not the citric acid-induced cough (11.7+/-1.8 vs. 12.8+/ 1.5; P>0.05). In contrast, exposure of guinea-pigs to bradykinin (10(-4) M) prior to the citric acid challenge resulted in a significant enhancement of the cough response (9.2+/-1.9 vs. 25.8+/-2.5; P<0.05) but not the bronchoconstriction (P>0.05). These data do not support a major peripheral role for substance P in the cough reflex, although bradykinin is able to sensitize the cough reflex. Furthermore, these data suggest that bronchoconstriction, induced by citric acid, is not responsible for the cough associated with this irritant. PMID- 15878718 TI - Evidence for the presence of P2y and P2x receptors with different functions in mouse stomach. AB - To clarify the function of P2 receptor subtypes in mouse stomach, the motor responses to ATP, alpha,beta-methyleneATP (alpha,beta-MeATP), P2X receptor agonist, 2-methylthioATP (2-MeSATP), P2Y receptor agonist, and the effects of the desensitisation of P2X receptors with alpha,beta-MeATP and of P2Y receptors with ADPbetaS were analysed recording the endoluminal pressure from whole-organ. ATP induced relaxation was antagonised by suramin, non-selective P2 receptor antagonist, by desensitisation of P2Y receptors with ADPbetaS, and increased by desensitisation of P2X receptors with alpha,beta-MeATP. alpha,beta-MeATP produced biphasic responses: relaxation, reduced by P2X- or P2Y desensitisation, and contraction, almost abolished by P2X desensitisation and potentiated by P2Y desensitisation. 2-MeSATP induced relaxation, which was antagonised by P2Y desensitisation and increased by P2X desensitisation. Tetrodotoxin increased the relaxation induced by purines and deeply antagonised the contraction to alpha,beta-MeATP. Our results suggest that in mouse stomach are present muscular P2Y receptors, subserving relaxation, and neuronal presynaptic P2X receptors, mediating contraction. PMID- 15878719 TI - gamma-Globulin-induced modulation with necrotic-like morphology of peripheral blood neutrophils. AB - To determine the effect of intravenous immunoglobulin-administration on neutrophil function, we obtained neutrophils from patients with an acute phase of Kawasaki disease. In vitro IgG-induced modulation of neutrophils into Annexin-V positive and propidium iodide-negative cells was observed in 20 of 28 patients in the presence of more than 300 microg/ml IgG and showed necrosis-like changes in morphologic features. However, we could not find any patients showing promotion of the sub-G1 cell fraction on DNA content analysis. The modulatory effect of in vitro IgG was not observed in neutrophils from healthy volunteers and was significantly correlated with the antifebrile effect of in vivo IgG. PMID- 15878720 TI - Histidine and carnosine delay diabetic deterioration in mice and protect human low density lipoprotein against oxidation and glycation. AB - In vivo effects of histidine and carnosine against diabetic deterioration in diabetic Balb/cA mice were studied. Histidine and carnosine at 0.5, 1 g/l were added into drinking water. After 4 weeks intake of these agents, the content of histidine and carnosine in plasma, heart and liver significantly elevated (P < 0.05). The intake of these agents significantly decreased plasma glucose and fibronectin levels (P < 0.05); however, only 1 g/l histidine and carnosine treatments significantly increased insulin level (P < 0.05) in diabetic mice. Triglyceride level in heart and liver was dose-dependently reduced by histidine or carnosine treatments (P < 0.05); however, only 1 g/l histidine and carnosine treatments significantly reduced cholesterol level in heart and liver (P < 0.05). The administration of histidine or carnosine significantly enhanced catalase activity and decreased lipid oxidation levels in kidney and liver (P < 0.05); however, only 1 g/l histidine and carnosine treatments significantly increased glutathione peroxidase activity (P < 0.05). The increased interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha in diabetic mice were significantly suppressed by the intake of histidine or carnosine (P < 0.05). In human low density lipoprotein, histidine or carnosine showed dose-dependently suppressive effect in glucose-induced oxidation and glycation (P < 0.05). These data suggest that histidine and carnosine are potential multiple-protective agents for diabetic complications prevention or therapy. PMID- 15878721 TI - Changes in ovarian steroidogenesis in insulin-resistant, type 2 diabetic Goto Kakizaki rats after thyroidectomy and gonadotropin treatment. AB - The present study used thyroidectomized insulin-resistant, type 2 diabetic Goto Kakizaki (GK) rats to assess whether insulin resistance and hypothyroidism modulate ovarian physiology. Animals were treated with daily injections of 5 IU equine chorionic gonadotropin for 5 days starting 1 week after thyroidectomy. Control groups included rats of GK and control (Wistar) strains treated only with equine chorionic gonadotropin or thyroidectomy, or with no treatment (intact). In Wistar rats, equine chorionic gonadotropin injections tended to increase the serum concentrations of luteinizing hormone (LH) and testosterone more in the thyroidectomy group than in intact rats. Similar changes in LH and testosterone were observed in the thyroidectomy + equine chorionic gonadotropin and equine chorionic gonadotropin groups of GK rats, but the LH and testosterone levels in the thyroidectomy + equine chorionic gonadotropin group were significantly higher in GK rats. Expression of ovarian LH receptor messenger RNA (mRNA) was enhanced by thyroidectomy. The LH receptor mRNA levels were significantly higher in the thyroidectomy+equine chorionic gonadotropin group of GK rats than in the corresponding group of control rats. These results indicate that hypothyroidism in animals with insulin resistance and type 2 diabetes promotes LH and testosterone secretions, and suggests that the enhanced-testosterone levels is partially mediated by the enhancement of LH receptor expression and an increase in the serum level of LH. PMID- 15878722 TI - Neuropsychology of timing and time perception. AB - Interval timing in the range of milliseconds to minutes is affected in a variety of neurological and psychiatric populations involving disruption of the frontal cortex, hippocampus, basal ganglia, and cerebellum. Our understanding of these distortions in timing and time perception are aided by the analysis of the sources of variance attributable to clock, memory, decision, and motor-control processes. The conclusion is that the representation of time depends on the integration of multiple neural systems that can be fruitfully studied in selected patient populations. PMID- 15878723 TI - Chronic treatment with haloperidol induces deficits in working memory and feedback effects of interval timing. AB - Normal participants (n=5) having no experience with antipsychotic drugs and medicated participants (n=5) with clinical experience with chronic low doses of haloperidol (3-10 mg/day for 2-4 months) in the treatment of neuroses were evaluated for the effects of inter-trial interval (ITI) feedback on a discrete trials peak-interval timing procedure. Feedback was presented during the ITI in the form of a histogram showing the distribution of the responses participants made on the previous trial plotted on a relative time scale. As feedback concerning the accuracy and precision of a reproduced duration (e.g., 7- and 14-s visual signals) became more remote in time, reproduced intervals gradually lengthened in duration. This rightward horizontal shift in peak time increased as a function of the probability of feedback and was enhanced by chronic treatment with haloperidol in a manner that was proportional to the duration of the signal. Our data suggest a gradual change in the underlying representation of the signal duration as a function of the remoteness of ITI feedback that is dependent upon both changes in working memory and the speed of the internal clock used to time durations in the seconds-to-minutes range. PMID- 15878724 TI - The effects of aging on time reproduction in delayed free-recall. AB - The experiments presented here demonstrate that normal aging amplifies differences in time production occurring in delayed free-recall testing. Experiment 1 compared the time production ability of two healthy aged groups as well as college-aged participants. During the test session, which followed a 24-h delay and omitted all feedback and examples of the two target intervals, the two groups of aged participants' over-produced a 6s interval. This effect is similar in form to errors shown by young participants, but twice the magnitude. Productions of a 17 s interval were generally accurate overall. However, further analysis indicated that the majority of aged participants over-produced the 17 s interval while a minority greatly under-produced the 17 s interval. Furthermore, aged participants showed violations of the scalar property of timing variability in the training session, while in the test session, only those who under-produced the 17 s interval showed this tendency. In contrast, training session performance was good for all participants. Experiments 2 and 3 investigated the ability of the participants in Experiment 1 to reproduce the length of a line from memory, under conditions analogous to those of the time production experiments. These experiments provided tests of the specificity of the errors observed in Experiment 1. Performance in the older participants was accurate, if more variable, compared to the young participants, in contrast to the time production results, indicating that production inaccuracy in free-recall is specific to interval timing in the current context. PMID- 15878725 TI - Effects of internal clock and memory disorders on duration reproductions and duration productions in patients with Parkinson's disease. AB - Patients with Parkinson's disease (PD) exhibit deficits in perceptual and motor timing as well as impairments in memory and attentional processes that are related to dysfunction of dopaminergic systems in the basal ganglia. The aim of the present study was to assess the relationships existing between impaired duration judgments and defective memory and attention in PD patients. We compared time performance of medicated PD patients and control subjects on a duration reproduction task that is highly memory-dependent, and on a duration production task that could reveal effects of changes in the speed of internal time-keeping mechanisms. Each task was performed in a control counting condition and in a condition requiring divided attention between the temporal task and a concurrent reading task. Moreover, PD patients and control subjects were assessed on memory and attention using a battery of neuropsychological tests. The results revealed that in the concurrent reading condition of the reproduction task, duration judgments tended to be more variable in PD patients than in control subjects. Moreover, variability of duration reproductions was correlated with both measures of memory and of disease severity. In the concurrent reading condition of the production task, duration judgments were significantly shorter in PD patients than in control subjects, and accuracy of duration productions was correlated with scores on sub-tests of short-term memory. The findings suggest that the administration of dopamine did not entirely remove the memory deficits in PD patients. Moreover, DA treatment would have abnormally accelerated the rate of the internal clock leading to shorter duration productions in PD patients. The whole results indicate that dopamine administration in patients might have overshadowed the slowing rate of the internal clock usually reported in non medicated PD patients, without entirely restoring all of the memory functions. PMID- 15878726 TI - Variable foreperiod deficits in Parkinson's disease: dissociation across reflexive and voluntary behaviors. AB - The effect of a visual warning signal (1.0-6.5 s random foreperiod, FP) on the latency of voluntary (hand-grip) and reflexive (startle-eyeblink) reactions was investigated in Parkinson's disease (PD) patients and in young and aged control subjects. Equivalent FP effects on blink were observed across groups. By contrast, FP effects diverged for voluntary responses across groups with no effect of foreperiod duration for PD patients. The convergence of these results with findings from animal research suggests that interval-timing processes associated with higher level voluntary behaviors are dependent upon intact dopaminergic pathways, while those associated with lower level reflexive behaviors are spared in PD. PMID- 15878727 TI - Effects of focal basal ganglia lesions on timing and force control. AB - Studies of basal ganglia dysfunction in humans have generally involved patients with degenerative disorders, notably Parkinson's disease. In many instances, the performance of these patients is compared to that of patients with focal lesions of other brain structures such as the cerebellum. In the present report, we studied the performance of patients with focal basal ganglia lesions on three fundamental motor tasks. The patients all had suffered unilateral damage in the striatum and were tested in the chronic state. The first task required the participants to tap with their index finger as fast as possible; this test provided a simple assessment of motor competence. Compared to controls, the maximum tapping rate was lower for the patients when tapping with their contralesional limb, although the deficit was not severe. The second and third tasks were designed to assess timing and force control, two functions that have been associated with basal ganglia function. The patients performed similar to controls on both tasks and showed no evidence of impairment when using their contralesional limb compared to their ipsilesional limb. The results indicate that unilateral basal ganglia lesions tend to produce minor motor problems in force control, and fail to support the hypothesized role of the basal ganglia in timing. PMID- 15878728 TI - Multidimensional sequence learning in patients with focal basal ganglia lesions. AB - Parkinson's patients have been found to be impaired in learning movement sequences. In the current study, patients with unilateral basal ganglia lesions due to stroke were tested on a serial reaction time task in which responses were based on the spatial location of each stimulus. The spatial locations either followed a fixed sequence or were selected at random, with learning operationalized as the difference in reaction time between these two conditions. In addition, three response-to-stimulus intervals were used, and these either followed a fixed sequence or were randomized. Compared to control participants, the patients showed normal learning of the spatial and temporal sequences, as well as normal cross-dimensional learning. This was true for performance with either the contralesional or ipsilesional hand. Sequence learning was not correlated with maximum tapping rate, a simple measure of motor impairment. These results raise questions concerning the use of Parkinson's disease as a model for studying basal ganglia dysfunction. PMID- 15878729 TI - Comparison of patients with Parkinson's disease or cerebellar lesions in the production of periodic movements involving event-based or emergent timing. AB - We have hypothesized a distinction between the processes required to control the timing of different classes of periodic movements. In one class, salient events mark successive cycles. For these movements, we hypothesize that the temporal goal is a requisite component of the task representation, what we refer to as event-based timing. In the other class, the successive cycles are produced continuously. For these movements, alternative control strategies can optimize performance, allowing timing to be emergent. In a previous study, patients with cerebellar lesions were found to be selectively impaired on event-based timing tasks; they were unimpaired on a continuously produced task. In the present study, patients with Parkinson's disease were tested on repetitive movement tasks in which timing was either event-based or emergent. Temporal variability on either type of task did not differ between on- and off-medication sessions for the Parkinson's patients nor did patient performance differ from that of controls. These results suggest that the basal ganglia play a minimal role in movement timing and that impairments on event-based timing tasks are specific to cerebellar damage. PMID- 15878730 TI - Eyeblink conditioning deficits indicate timing and cerebellar abnormalities in schizophrenia. AB - Accumulating evidence indicates that individuals with schizophrenia manifest abnormalities in structures (cerebellum and basal ganglia) and neurotransmitter systems (dopamine) linked to internal-timing processes. A single-cue tone delay eyeblink conditioning paradigm comprised of 100 learning and 50 extinction trials was used to examine cerebellar timing circuits in 13 medicated patients with schizophrenia and 13 age- and sex-matched controls. Patients with schizophrenia showed impaired learning of the conditioned response compared to controls and also greater within-subject variability in the timing of their responses. These findings are consistent with models of schizophrenia in which timing deficits underlie information-processing abnormalities and clinical features of the disorder. PMID- 15878731 TI - Interval-timing deficits in individuals at high risk for schizophrenia. AB - A duration-bisection procedure was used to study the effects of signal modality and divided attention on duration classification in participants at high genetic risk for schizophrenia (HrSz), major affective disorder (HrAff), and normal controls (NC). Participants learned short and long target durations during training and classified probe durations during test. All groups classified visual signals as shorter than equivalent duration auditory signals. However, the difference between auditory and visual signal classification was significantly larger for the HrSz group than for the NC group. We posit a model in which there is a clock rate difference between auditory and visual signals due to an attentional effect at the level of a mode switch that gates pulses into an accumulator. This attentionally mediated clock rate difference was larger for the HrSz participants than for the NC participants, resulting in a larger auditory/visual difference for the HrSz group. PMID- 15878732 TI - Auditory/visual duration bisection in patients with left or right medial-temporal lobe resection. AB - Patients with unilateral (left or right) medial temporal lobe lesions and normal control (NC) volunteers participated in two experiments, both using a duration bisection procedure. Experiment 1 assessed discrimination of auditory and visual signal durations ranging from 2 to 8 s, in the same test session. Patients and NC participants judged auditory signals as longer than equivalent duration visual signals. The difference between auditory and visual time discrimination was equivalent for the three groups, suggesting that a unilateral temporal lobe resection does not modulate the modality effect. To document interval-timing abilities after temporal lobe resection for different duration ranges, Experiment 2 investigated the discrimination of brief, 50-200 ms, auditory durations in the same patients. Overall, patients with right temporal lobe resection were found to have more variable duration judgments across both signal modality and duration range. These findings suggest the involvement of the right temporal lobe at the level of the decision process in temporal discriminations. PMID- 15878733 TI - About hemispheric differences in the processing of temporal intervals. AB - The purpose of the present study was to identify differences between cerebral hemispheres for processing temporal intervals ranging from .9 to 1.4 s. The intervals to be judged were marked by series of brief visual signals located in the left or the right visual field. Series of three (two standards and one comparison) or five intervals (four standards and one comparison), marked by sequences of 4 or 6 signals, were compared. While discrimination, as estimated by d', was significantly better in the 4-standard than in the 2-standard condition when stimuli were presented in the left visual field (LVF), this number-of standard effect on discrimination varied with the difficulty levels when the signals were presented in the LVF. Moreover, the discrimination levels were constant for the different base durations with stimuli presented in the LVF, but not with stimuli presented in the right visual field. This article discusses the implication of these findings for the study of hemispheric dominance for temporal processing and for a single-clock hypothesis. PMID- 15878734 TI - Auditory discrimination of anisochrony: influence of the tempo and musical backgrounds of listeners. AB - This study explored the influence of several factors, physical and human, on anisochrony's thresholds measured with an adaptive two alternative forced choice paradigm. The effect of the number and duration of sounds on anisochrony discrimination was tested in the first experiment as well as potential interactions between each of these factors and tempo. In the second experiment, the tempo or the inter onset interval (IOI) was varied systematically between 80 and 1000 ms. The results showed that just noticeable differences increase linearly and proportionally with IOI in accordance with Weber's law except for quickest tempo (IOI of 80 ms). The third experiment investigated the role of musical training on anisochrony thresholds obtained for different IOI. It focused on differential effects of musical experiences by comparing non-musicians, instrumentalists, and percussionists thresholds. The results of the present study replicated the findings of previous experiments regarding the adequacy of Weber's law for slow rhythm and provided evidence for its departure for fast tempos. Moreover, thresholds from percussionists seem distinguishable from the ones of other listeners by their highest sensitivity to temporal shifts suggesting therefore the necessity to control the nature of musical experiences. The results are discussed according to current models of time perception. PMID- 15878735 TI - Stability of RNA isolated from human trabecular bone at post-mortem and surgery. AB - To determine the reliability of gene expression studies in human post-mortem bone, it is important to evaluate the stability of RNA isolated from such tissues as a function of the post-mortem interval. The stability of total RNA and bone specific mRNA species was examined in bone samples obtained from routine autopsies and at surgery. The optimal temperature for any storage and transport of the bone before RNA isolation was shown to be 4 degrees C, and RT-PCR analysis is the preferred technique for the analysis of gene expression in post-mortem bone as it tolerates partial RNA degradation. For gene expression studies in bone, post-mortem cases, with a post-mortem interval of less than 48 h, should be selected, and the time that bone is stored after retrieval at autopsy or surgery should be kept to a minimum. Overall, our findings indicate that with appropriate storage and handling, RNA can be reliably isolated from human bone obtained at post-mortem and surgery to study ex vivo the pattern of gene expression in healthy individuals and in patients with musculoskeletal diseases such as osteoporosis and osteoarthritis. PMID- 15878736 TI - Homocysteine attenuates the expression of osteocalcin but enhances osteopontin in MC3T3-E1 preosteoblastic cells. AB - It has been pointed out that very high plasma levels of homocysteine are characteristic of homocystinuria, a rare autosomal recessive disease accompanied by the early onset of generalized osteoporosis. However, it is unclear by which mechanism hyperhomocysteine induces osteoporosis, although it is known to interfere with the formation of cross-links in collagen, an essential process in bone formation. Therefore, we investigated the effect of homcysteine on expression of osteocalcin and osteopontin in MC3T3-E1 preosteoblastic cells. Confluent cells were grown in RPMI 1640 containing 10% fetal calf serum with or without homocysteine in an atmosphere of 95% humidified air, 5% CO2 at 37 degrees C. The secretion of osteocalcin from the cells increased time-dependently until the end of culture (day 34), but 500 microM homocysteine led to an approximately 61% decrease for osteocalcin after 19 days of culture as compared with the control. On the other hand, osteopontin was not inhibited by 500 microM homocysteine but rather activated, and ranged from 134%-209% of the control level in the period from 10 days until the end of culture. From analysis of RT-PCR for mRNA of osteocalcin and osteopontin at the end of the culture, homocysteine levels of 100 and 500 microM significantly increased the expression of osteopontin mRNA with the control (p < 0.05). In contrast, the expression of osteopontin mRNA was suppressed in a dose-dependent manner, showing a mirror image of the effect on osteopontin mRNA. These findings suggest that hyperhomocystenemia appears to be an independent risk factor for osteoporosis by disturbing osteoblast function. PMID- 15878737 TI - Erythropoietin hypersensitivity in primary familial and congenital polycythemia: role of tyrosines Y285 and Y344 in erythropoietin receptor cytoplasmic domain. AB - Erythropoietin receptor (EPOR) gene mutations leading to truncations of the cytoplasmic, carboxy-terminal region of EPOR have been described in some patients with primary familial and congenital polycythemia (PFCP), a disorder characterized by isolated erythrocytosis and increased sensitivity of erythroid progenitors to Epo. We studied the role of EPOR in the pathogenesis of PFCP and the requirement for intracytoplasmic tyrosine residues Y285 and Y344 in generation of Epo hypersensitivity phenotype. Interleukin-3-dependent hematopoietic cells were engineered to express variant human EPORs using retrovirus-mediated gene transfer. We introduced tyrosine to phenylalanine substitutions in EPOR-ME, a naturally occurring, mutant human EPOR (G5881T), truncated by 110 carboxy-terminal amino acids and associated with autosomal dominantly inherited PFCP. Cells expressing EPOR-ME exhibited increased Epo sensitivity compared to cells expressing wild type EPOR. Mutation of Y285 alone had a relatively minor effect on Epo hypersensitivity whereas mutation of Y344 resulted in loss of increased Epo sensitivity. Expression of a tyrosine-null truncated EPOR conferred further decrease of Epo-mediated proliferation suggesting that both Y285 and Y344 may contribute to proliferation signals. In the context of EPOR-ME, Y344 was required for Epo-induced Stat5 tyrosine phosphorylation. The positive effect of either Y285 or Y344 on cellular proliferation was associated with Epo-induced tyrosine phosphorylation of Stat1. These findings suggest that both tyrosine residues Y285 and Y344 in the cytoplasmic domain of EPOR-ME may contribute to increased Epo sensitivity that is characteristic of PFCP phenotype. PMID- 15878738 TI - Altered renal elimination of organic anions in rats with chronic renal failure. AB - The progress of chronic renal failure (CRF) is characterized by the development of glomerular and tubular lesions. However, little is known about the expression of organic anions renal transporters. The objective of this work was to study, in rats with experimental CRF (5/6 nephrectomy), the expression of the organic anion transporter 1 (OAT1) and organic anion transporter 3 (OAT3) and their contribution to the pharmacokinetics and renal excretion of p-aminohippurate (PAH). Two groups of animals were used: Sham and CRF. Six months after surgery, systolic blood pressure and plasma creatinine concentrations were significantly higher in CRF groups. CRF rats showed a diminution in: the filtered, secreted and excreted load of PAH; the systemic clearance of PAH; the renal OAT1 expression; and the renal Na-K-ATPase activity. No remarkable modifications were observed in the OAT3 expression from CRF kidneys. The diminution in the systemic depuration and renal excretion of PAH may be explained by the decrease in its filtered and secreted load. The lower OAT1 expression in remnant renal mass of CRF rats or/and the lower activity of Na-K-ATPase might justify, at least in part, the diminished secreted load of this organic anion. PMID- 15878739 TI - Budding, vesiculation and permeabilization of phospholipid membranes-evidence for a feasible physiologic role of beta2-glycoprotein I and pathogenic actions of anti-beta2-glycoprotein I antibodies. AB - The in vivo physiologic role of beta2-glycoprotein I (beta2GPI) is presumed to be related to its interactions with negatively charged phospholipid membranes. Increased quantities of procoagulant microparticles derived by the vesiculation of blood cells have been detected in patients with antiphospholipid syndrome (APS) frequently associated with antibodies against beta2GPI (anti-beta2GPI). We investigated the influence of beta2GPI and anti-beta2GPI on giant phospholipid vesicles (GPVs). GPVs composed of phosphatidylserine and phosphatidylcholine were formed in an aqueous medium and individually transferred to a compartment containing either beta2GPI, anti-beta2GPI, or beta2GPI along with anti-beta2GPI. Shape changes of a single GPV were observed by a phase contrast microscope. Most GPVs transferred to the solution containing only beta2GPI budded moderately. Upon the transfer of GPVs to the solution containing beta2GPI and anti-beta2GPI either from patient with APS or mouse monoclonal anti-beta2GPI Cof-22, the budding was much more pronounced, generating also daughter vesicles. No such effects were seen when GPV was transferred to the solution containing anti-beta2GPI without beta2GPI. Our results suggest a significant physiologic role of beta2GPI in the budding of phospholipid membranes, which may be explained by the insertion of the C-terminal loop of beta2GPI into membranes, thus increasing the surface of the outer layer of a phospholipid bilayer. Anti-beta2GPI, recognizing domains I to IV of beta2GPI, enhanced the budding and vesiculation of GPVs in the presence of beta2GPI. This might be a novel pathogenic mechanism of anti-beta2GPI, promoting in vivo the expression of proadhesive and procoagulant phospholipid surfaces in APS. PMID- 15878740 TI - Dermatan sulfate is the predominant antithrombotic glycosaminoglycan in vessel walls: implications for a possible physiological function of heparin cofactor II. AB - The role of different glycosaminoglycan species from the vessel walls as physiological antithrombotic agents remains controversial. To further investigate this aspect we extracted glycosaminoglycans from human thoracic aorta and saphenous vein. The different species were highly purified and their anticoagulant and antithrombotic activities tested by in vitro and in vivo assays. We observed that dermatan sulfate is the major anticoagulant and antithrombotic among the vessel wall glycosaminoglycans while the bulk of heparan sulfate is a poorly sulfated glycosaminoglycan, devoid of anticoagulant and antithrombotic activities. Minor amounts of particular a heparan sulfate (< 5% of the total arterial glycosaminoglycans) with high anticoagulant activity were also observed, as assessed by its retention on an antithrombin-affinity column. Possibly, this anticoagulant heparan sulfate originates from the endothelial cells and may exert a significant physiological role due to its location in the interface between the vessel wall and the blood. In view of these results we discuss a possible balance between the two glycosaminoglycan-dependent anticoagulant pathways present in the vascular wall. One is based on antithrombin activation by the heparan sulfate expressed by the endothelial cells. The other, which may assume special relevance after vascular endothelial injury, is based on heparin cofactor II activation by the dermatan sulfate proteoglycans synthesized by cells from the subendothelial layer. PMID- 15878741 TI - Identification of novel mutations of the human N-acetylglutamate synthase gene and their functional investigation by expression studies. AB - The mitochondrial enzyme N-acetylglutamate synthase (NAGS) produces N acetylglutamate serving as an allosteric activator of carbamylphosphate synthetase 1, the first enzyme of the urea cycle. Autosomal recessively inherited NAGS deficiency (NAGSD) leads to severe neonatal or late-onset hyperammonemia. To date few patients have been described and the gene involved was described only recently. In this study, another three families affected by NAGSD were analyzed for NAGS gene mutations resulting in the identification of three novel missense mutations (C200R [c.598T > C], S410P [c.1228T > C], A518T [c.1552G > A]). In order to investigate the effects of these three and two additional previously published missense mutations on enzyme activity, the mutated proteins were overexpressed in a bacterial expression system using the NAGS deficient E. coli strain NK5992. All mutated proteins showed a severe decrease in enzyme activity providing evidence for the disease-causing nature of the mutations. In addition, we expressed the full-length NAGS wild type protein including the mitochondrial leading sequence, the mature protein as well as a highly conserved core protein. NAGS activity was detected in all three recombinant proteins but varied regarding activity levels and response to stimulation by l-arginine. In conclusion, overexpression of wild type and mutated NAGS proteins in E. coli provides a suitable tool for functional analysis of NAGS deficiency. PMID- 15878742 TI - The type of collagen cross-link determines the reversibility of experimental skin fibrosis. AB - Fibrotic processes in humans are characterised by an excessive accumulation of collagen containing increased levels of hydroxyallysine-derived cross-links. The occurrence of these cross-links appears to be an important criterion in assessing the irreversibility of fibrosis. We hypothesise that increased hydroxyallysine cross-linking results in a collagenous matrix that is less susceptible to proteolytic degradation and therefore the collagen deposition is no longer reversible. In this report, we show that collagen matrices with increased hydroxyallysine cross-link levels were less susceptible to matrix metalloproteinase 1 degradation than are collagen matrices containing low hydroxyallysine levels. These data indicate that the type of collagen cross-link influences collagen catabolism. In vivo evidence for the importance of the cross linking type in determining the reversibility of the fibrotic process was found using the bleomycin-induced skin fibrosis mouse model. The analysis of the accumulated collagen in the fibrotic skin of bleomycin-treated mice did not reveal an increase in hydroxyallysine cross-link levels. In concurrence with our hypothesis, the collagen accumulation resolved in time when the mice were no longer receiving bleomycin treatment, showing the reversibility of the fibrosis. In conclusion, our data indicate that the type of collagen cross-linking is an important factor in determining whether the outcome of the fibrotic process is reversible or not. PMID- 15878743 TI - Oxidative stress in patients with phenylketonuria. AB - Phenylketonuria (PKU) is an autossomal recessive disease caused by phenylalanine 4-hydroxylase deficiency, which is a liver-specific enzyme that catalyzes the hydroxylation of l-phenylalanine (Phe) to l-tyrosine (Tyr). The deficiency of this enzyme leads to the accumulation of Phe in the tissues and plasma of patients. The clinical characterization of this disease is mental retardation and other neurological features. The mechanisms of brain damage are poorly understood. Oxidative stress is observed in some inborn errors of intermediary metabolism owing to the accumulation of toxic metabolites leading to excessive free radical production and may be a result of restricted diets on the antioxidant status. In the present study we evaluated various oxidative stress parameters, namely thiobarbituric acid-reactive species (TBA-RS) and total antioxidant reactivity (TAR) in the plasma of PKU patients. The activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also measured in erythrocytes from these patients. It was observed that phenylketonuric patients present a significant increase of plasma TBA-RS measurement, indicating a stimulation of lipoperoxidation, as well as a decrease of plasma TAR, reflecting a deficient capacity to rapidly handle an increase of reactive species. The results also showed a decrease of erythrocyte GSH-Px activity. Therefore, it is presumed that oxidative stress is involved in the pathophysiology of the tissue damage found in PKU. PMID- 15878744 TI - Islet amyloid polypeptide gene promoter polymorphisms are not associated with Type 2 diabetes or with the severity of islet amyloidosis. AB - The over-expression of the islet amyloid polypeptide (IAPP) gene could be a causal factor for islet amyloidosis and beta-cell destruction in Type 2 diabetes (T2DM). An IAPP gene promoter polymorphism, IAPP-132G to A, has been associated with T2DM in Spain. To investigate this polymorphism in other cohorts and in relation to therapy, DNA from 425 T2DM and 279 unrelated, non-diabetic UK subjects (ND) and 102 T2DM and 80 ND Finnish subjects was examined. The relationship of amyloid severity (percent amyloid/islet) to prevalence (number of islets affected) and the association of IAPP-132G/A with amyloid was determined in post-mortem pancreas from 38 T2DM subjects. The -132G/A was not associated with T2DM in the UK cohorts (4.5% T2DM; 3.2% ND) or associated with requirement for insulin therapy by 6 years. The mutation was and undetected in the Finnish samples but a new variant, -166T/C, was identified in 2 Finnish T2DM subjects. 132G/A was found in 2/38 diabetic, amyloid-containing and 3/19 ND, amyloid-free subjects. The islet amyloid severity was linearly correlated with the prevalence in T2DM. The IAPP-132G/A promoter polymorphism is not associated with T2DM, a requirement for insulin therapy or with the degree of islet amyloidosis in cohorts from the UK or Finland. PMID- 15878745 TI - Altered expression of iron transport proteins in streptozotocin-induced diabetic rat kidney. AB - Diabetes mellitus is associated with altered iron homeostasis in both human and animal diabetic models. Iron is a metal oxidant capable of generating reactive oxygen species (ROS) and has been postulated to contribute to diabetic nephropathy. Two proteins involved in iron metabolism that are expressed in the kidney are the divalent metal transporter, DMT1 (Slc11a2), and the Transferrin Receptor (TfR). Thus, we investigated whether renal DMT1 or TfR expression is altered in diabetes, as this could potentially affect ROS generation and contribute to diabetic nephropathy. Rats were rendered diabetic with streptozotocin (STZ-diabetes) and renal DMT1 and TfR expression studied using semi-quantitative immunoblotting and immunofluorescence. In STZ-diabetic Sprague Dawley rats, renal DMT1 expression was significantly reduced and TfR expression increased after 2 weeks. DMT1 downregulation was observed in both proximal tubules and collecting ducts. Renal DMT1 expression was also decreased in Wistar rats following 12 weeks of STZ-diabetes, an effect that was fully corrected by insulin-replacement but not by cotreatment with the aldose reductase inhibitor, sorbinil. Increased renal TfR expression was also observed in STZ-diabetic Wistar rats together with elevated cellular iron accumulation. Together these data demonstrate renal DMT1 downregulation and TfR upregulation in STZ-diabetes. Whilst the consequence of altered DMT1 expression on renal iron handling and oxidant damage remains to be determined, the attenuation of the putative lysosomal iron exit pathway in proximal tubules could potentially explain lysosomal iron accumulation reported in human diabetes and STZ-diabetic animals. PMID- 15878746 TI - Partial characterisation of the human GFAT promoter: effect of single nucleotide polymorphisms on promoter function. AB - The 5'-flanking region of the human glutamine:fructose-6-phosphate amidotransferase (GFAT) gene was characterised as a functional active promoter and the GFAT gene contained multiple transcription start sites. A novel single nucleotide polymorphism identified at position -1412 (G to C) had a functional effect on promoter activity and EMSA revealed specific binding of nuclear proteins to this region. PMID- 15878748 TI - Are psycho-social and behavioural factors related to health related-quality of life in patients with head and neck cancer? A systematic review. AB - The survival rate in advanced cancer of the head and neck has remained at approximately 50%, and efforts are now directed towards reducing the impact of the disease and its treatment in terms of functioning and health related-quality of life (HR-QoL). Factors such as stage, site of disease and type of treatment all impact on HR-QoL, but it is unclear what additional factors influence HR-QoL. A systematic review was undertaken of studies that have investigated psycho social or behavioural factors associated with HR-QoL in this patient group. Literature was systematically searched using electronic databases and hand searching relevant journals. Data were sought on HR-QoL and studies were only included if the measurement instrument was recognised as a reliable and valid measure of HR-QoL. Studies had to include at least one psycho-social or behavioural predictor variable. Sixteen studies fulfilling the inclusion criteria were identified and reviewed. Five main factors were associated with varying degrees with HR-QoL, personality, social support, satisfaction with consultation and information, behavioural factors, such as consuming alcohol and smoking, and depressive symptoms. The major difficulty with synthesising the findings was the amount of different indices of QoL that have been used. However, a number of psycho-social factors have been investigated in relation to HR-QoL in head and neck cancer patients, some of which are potentially modifiable, such as those related to informational needs. Further research is needed to investigate other psychological factors which may influence aspects of HR-QoL. By understanding the relationship between HR-QoL and potentially modifiable variables, interventions can be designed with the aim of improving a patient's long-term well-being. PMID- 15878749 TI - Association between polymorphisms of the GPX1 gene and second primary tumours after index squamous cell cancer of the head and neck. AB - We investigated the association between genetic polymorphisms in GPX1 gene amongst patients who had index squamous cell carcinoma (SCCHN) and a second primary tumour (SPT) after a primary SCCHN in a case-control study. GPX1 genotypes were determined for 61 patients with SPT and for 259 control subjects by a PCR technique using a fluorescent-labelled primer. Analysis was by an ABI automated fluorescent sequencer. The associations between specific genotypes and the development of SPT were examined by logistic regression. A significant difference was found between the control group and the SPT cases in allele frequencies of GPX1 ALA( *)6 and ALA( *)7 (p(trend)=0.04). These results suggest that polymorphisms in the GPX1 gene may be a marker for SPT development and further studies are indicated. PMID- 15878750 TI - CD40 function in squamous cell cancer of the head and neck. AB - CD40 is expressed on basal keratinocytes and Squamous Cell Cancer of the Head and Neck (SCCHN) tumor cells in vivo and in vitro. CD40 ligation reduces proliferation of SCCHN cell lines and enhances EGFr mediated inhibition of proliferation. We investigated the mechanisms of CD40 function and EGFr cross communication in SCCHN cell lines. CD40 ligation inhibited spontaneous and Fas induced apoptosis. CD40 ligation specifically increased the secretion of IL-8, VEGF and PGE(2) but not IL-6, IL-10, FasL, GM-CSF, or TGFalpha. Co-ligation with EGFr further increased IL-8, VEGF and PGE(2) secretion. CD40 ligation also induced delayed activation and tyrosine phosphorylation of EGFr. CD40 induces secretion of specific proinflammatory and proangiogenic cytokines, inhibits spontaneous and Fas-induced apoptosis and increases EGFr phosphorylation. CD40 signaling may enhance the survival of SCCHN and tumor stroma. PMID- 15878751 TI - Incidence rates and trends for oral and pharyngeal cancer in North Carolina: 1990 1999. AB - This study describes the incidence of oral and pharyngeal cancer (OPC) in North Carolina (NC) from 1990 to 1999. OPC cases 1990-1999 from the NC Central Cancer Registry were included. Variables included date of diagnosis, county of residence, anatomical site, histology, age at diagnosis, sex, race, stage at diagnosis, grade, tumor behavior, and laterality. The overall average annual rate of OPC was 10.6 cases per 100,000 population. OPC incidence was highest for African-American males and for eastern NC; there was no significant trend in the overall annual incidence rate. Squamous cell carcinoma of the lip increased for white females and other oral cancers decreased among all females. With a few specific exceptions, NCCCR rates were similar to SEER rates. North Carolina continues to have substantial OPC incidence and substantial racial disparities persist. PMID- 15878752 TI - Immunohistochemical expression of RANK, GRalpha and CTR in central giant cell granuloma of the jaws. AB - The aim was to evaluate the phenotypic expression of various cellular osteotropic factors in central giant cell granuloma (CGCG). Paraffin-embedded tissue from 27 aggressive and 10 non-aggressive cases of CGCG was assessed for the expression of RANK, GRalpha and CTR using immunohistochemistry. In addition, a staining intensity-distribution (SID) score (proportion of stained cells x staining intensity) was used to assess immunoreactivity of each marker. The results showed that the multinucleated giant cells (MGC), mononuclear stromal cells (MSC) and endothelial cells were intensely positives for GRalpha, moderate for RANK and weak-to-moderate for CTR in all clinical groups, whereas spindle-shaped cells were intensely immunoreactive to GRalpha and unreactive to CTR and RANK. Although neither difference in RANK and GRalpha expression nor the SID score between the clinical forms of CGCG was observed, a statistically significant difference for CTR was evident. Furthermore, the comparison of the marker expression and SID score showed a significant correlation for all three markers within the clinical groups, except for GRalpha in the non-aggressive lesions where a weak and no significant correlation was detected. It was concluded that although the MGC share some similarities with the osteoclasts, they demonstrate phenotypic differences from each other that suggest a distinct precursor. The expression of RANK, GRalpha and CTR also suggest a role for these receptors in the resorptive activity of different cellular groups in CGCG and may lead to a more effective use of therapeutic inhibitors of bone resorption for the treatment of these disorders. PMID- 15878753 TI - Reliability of histologic diagnosis of clinically normal intraoral tissue adjacent to clinically suspicious lesions in former upper aerodigestive tract cancer patients. AB - Oral lesion biopsies often include adjacent clinically normal tissue to assess lesion spread, but the reliability of adjacent tissue histologic diagnoses has not been reported. Sixty-seven patients presenting with subsequent suspicious intraoral lesions underwent tissue staining with tolonium chloride. Biopsies of unstained tissue adjacent to suspicious lesions were reviewed by independent pathologists. The weighted kappa (kappa(w)) for histologic classification was 0.75 (95% confidence interval [CI] 0.64, 0.86) and was lower for: inflamed specimens [kappa(w)=-0.13 (95% CI -0.30, 0.05)] versus noninflamed specimens [kappa(w)=0.81 (95% CI 0.69, 0.92)], wedge biopsies [kappa(w)=0.58 (95% CI 0.37, 0.79)] versus punch biopsies [kappa(w)=0.90 (95% CI 0.83, 0.97)], and current smokers [w=0.59 (95% CI 0.32, 0.86)] versus past and never smokers [kappa(w)=0.82 (95% CI 0.72, 0.91)]. Histologic diagnosis of subsets of clinically normal oral specimens adjacent to suspicious tissue has low reliability. Molecular techniques may improve diagnoses in these subsets. PMID- 15878754 TI - Vascular endothelial growth factor gene-460 C/T polymorphism is a biomarker for oral cancer. AB - Oral cancer is one of the most common malignant diseases in Taiwan. The incidence of male oral cancer is 9.01 times than that of female. The formation or progression of oral cancer may be associated with a polymorphism of the vascular endothelial growth factor (VEGF) gene. The most frequently seen polymorphism is BstUI (C to T) located at the -460th nucleotide upstream of the VEGF gene. We investigated whether this polymorphism could be a genetic marker of oral cancer. A normal control group of 230 healthy people and 137 patients with oral cancer were examined. The polymorphism was detected by polymerase chain reaction-based restriction analysis. The analysis revealed significant differences between normal individuals and patients with cancer (P<0.001). The distribution of the "TT" homozygote in the patient group was greater than that in the control group. The odds ratio per copy of the "T/C" ratio was 9.62 (95% confidence interval 5.81 15.87), which means that for a group of people with a higher T/C ratio have higher risk in getting oral cancer. There is no gender difference in this VEGF gene polymorphism. Therefore, the BstUI polymorphism of the VEGF gene is a suitable genetic marker of oral cancer. PMID- 15878755 TI - Profiling EGFR activity in head and neck squamous cell carcinoma by using a novel layered membrane Western blot technology. AB - Given the role of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinomas (HNSCC), several rational approaches have now been utilized to abrogate tyrosine kinase activity and its disengagement from downstream signal transducers. Monitoring the activity of these molecules could potentially be useful to determine not only drug efficacy but also to identify HNSCC patients most likely to benefit from this type of therapy. In this study we have used a novel high throughput multi-layered Western blotting (MLWestern) method that allows the detection of multiple proteins from a single experiment in order to characterize key components in the EGFR signaling pathway in HNSCC cells. Total and activated forms of EGFR and the downstream effectors, Erk and Akt were readily detected in HNSCC cells, where in the control cells (HaCaT) these proteins could only be detected in EGF stimulated cells. Results from conventional Western blot and MLWestern were comparable. Clustering analysis of protein expression revealed similarities in cellular response between some of the cell lines indicative of similarities in their biological response. The data indicate that MLWestern can be potentially applied to identify molecular targets that could be used for rational therapeutic intervention strategies. PMID- 15878756 TI - HGF is released from buccal fibroblasts after smokeless tobacco stimulation. AB - To investigate the effect of smokeless tobacco (ST) on (1) HGF, KGF and GM-CSF expression by buccal fibroblasts and (2) on keratinocyte and fibroblast proliferation. Buccal fibroblasts were stimulated with different concentrations of ST extracts in a double dilution from 0.50% w/v to 0.03% w/v. Supernatant was collected after 24/48/72/96 h and assayed for HGF, KGF, and GM-CSF by ELISA. The amount of RNA was used as an indicator of fibroblast cell number. Buccal epithelial cell proliferation was determined by CyQUANT proliferation assay. The amount of HGF and KGF in the supernatant was dependent on exposure time and on concentration of the tobacco extract. High concentration increased production of HGF 4-fold. KGF production was doubled when high concentration of tobacco was used, low concentration did not stimulate cells. GM-CSF production was low in both stimulated and non-stimulated cells. Keratinocytes and fibroblasts showed no increase in proliferation after stimulation with increased concentrations of ST. The results suggest that HGF and KGF may play an important role as a paracrine growth factor in epithelial hyperplasia in ST lesions. PMID- 15878757 TI - Association between GSTT-1 gene deletion and the susceptibility to oral squamous cell carcinoma in cigarette-smoking subjects. AB - Oral squamous cell carcinoma (OSCC) is related to tobacco use and alcohol consumption and the individual susceptibility for development of this tumor has been associated with xenobiotic-metabolizing enzymes polymorphisms. Glutathione S transferase Theta-1 (GSTT1) is a phase II metabolic enzyme that catalyzes detoxification reactions of carcinogenic compounds. A genetic polymorphism of the GSTT1 gene is characterized by a complete gene deletion and consequent absence of the enzyme. The null genotype (gene deleted) of GSTT1 has a decreased capacity in detoxifying carcinogens present in tobacco smoke, leading the formation of DNA adducts and DNA damage. The purpose of the present study was to evaluate the association between GSTT1 polymorphism and risk for OSCC development in a Brazilian population. We investigated the GSTT1 polymorphism in 87 patients with OSCC and 81 age-sex matched controls. The genotypes were studied by PCR-based methods. Individuals homozygous for the wild type GSTT1 (+/+) and heterozygous (+/0) were grouped together. The prevalence of GSTT1 deficiency (null) was significantly higher in the group with OSSC patients with oral cancer of the floor of the mouth. The results suggest that GSTT1 polymorphism increases the risk for OSCC development. PMID- 15878758 TI - Treatment results of stage I oral tongue cancer with definitive radiotherapy. AB - The medical records of 133 patients with Stage I tongue cancer treated by definitive radiotherapy between 1966 and 2001 were reviewed. Overall survival rate (OS), progression free survival rate (PFS), and survival rate after recurrence were calculated according to the Kaplan-Meier method. We investigated prognostic factors for local control and risk factors of late neck LN metastasis. The 5-year OS was 81.8% and the 5-year PFS was 67.2%. The 5-year OS after local recurrence was 100% by salvage operation, and that after neck LN metastasis was 40.7% despite radical neck dissection. Tumor thickness over 5 mm and treatment without interstitial irradiation were prognostic factors for local control. Tumor diameter over 15 mm and tumor thickness over 5 mm were risk factors of late neck LN metastasis. We should consider prophylactic treatment for neck LN for high risk patients with Stage I tongue cancer in order to improve treatment results further. PMID- 15878759 TI - Neoadjuvant accelerated chemotherapy followed by hyperfractionated radiation therapy in patients with operable, locally advanced head and neck carcinoma. AB - A prospective phase II trial was carried out to evaluate an accelerated chemotherapy (CT) regimen followed by hyperfractionated radiation therapy (RT) in previously untreated Stage III-IV, operable (total laryngectomy), head and neck cancer patients. The current study evaluates overall survival, loco-regional control, organ preservation rates and toxicity. Between April 1997 and December 2002, 68 patients with advanced head and neck cancer were treated with 3 cycles of induction CT (cisplatin and 5-fluorouracil; days 1, 14, and 28) followed by hyperfractionated RT (7440 cGy/62 fractions). Sixty patients received the planned RT-CT treatment. Two months after the end of RT, 96% of patients had a clinical complete remission of the primary and 66% of the neck disease. At a median follow up of 32 months, the 3-year overall and disease-free survival rates were 66% and 76%, respectively. Seven patients recurred on the primary site, 1 on the neck and 2 patients only had distant metastases. The organ preservation rate was 73%. Acute grade 3-4 mucositis occurred in 75% of patients and an 18% rate of CT related cardiotoxicity was reported. The accelerated CT-RT regimen achieves a high rate of larynx preservation albeit with considerable toxicity. The current prospective clinical trial was approved by the Ethics Committee of the Centro di Riferimento Oncologico (C.R.O.) on May 27, 1996, # CRO-02-96. Written informed consent was required from all patients entering the study. PMID- 15878760 TI - Predictive factors for distant metastasis from oral and oropharyngeal squamous cell carcinoma. AB - The presence of distant metastasis after the initial treatment of head and neck squamous cell carcinoma is not considered a common event and is associated with a poor outcome. The objective of this study was to investigate the prevalence and risk factors associated with the diagnosis of distant metastasis in oral and oropharyngeal carcinoma patients. The medical charts of 2327 patients treated from 1954 to 1997 were reviewed. They were 1703 patients (73.2%) with oral cavity and 624 (26.8%), oropharyngeal tumours. Regarding the primary treatment: 637 patients (27.4%) underwent surgery alone; 1147 (49.3%), radiotherapy alone and 543 (23.3%), combined treatment (surgery and radiotherapy). During the follow-up period after the initial treatment, 89 patients (3.8%) were diagnosed with distant metastasis. The variables associated with the distant metastasis-free survival were: tumour site (p=0.008); T stage (p<0.001); N stage (p<0.001); treatment performed (p<0.001) and decade of admission at the institution (p<0.001). The multivariate analysis (Cox regression) showed that the clinical stage (p=0.007); treatment performed (p=0.012) and decade of admission at the institution (p=0.004) were independent predictive factors for distant metastasis. Distant metastasis has been diagnosed more frequently in the latter decade and the significant predictors associated with its presence were the advanced clinical stage and patients who had undergone combined treatment. PMID- 15878761 TI - Heparanase gene and protein expression in ameloblastoma: possible role in local invasion of tumor cells. AB - Ameloblastoma is the most common odontogenic neoplasm, particularized by its local invasiveness. Heparanase is the endo-glucuronidase enzyme that specifically cleaves heparan sulfate, the important modulator of extracellular matrix, and related to invasion of tumor cells. In this study, we addressed to show the gene expression and localization of heparanase in ameloblastoma. Immunohistochemistry and in situ hybridization of heparanase were carried out in 23 ameloblastomas. Strong expression of heparanase at both mRNA and protein levels was detected in all ameloblastomas studied. Small tumor nests and budding epithelial branches showed stronger staining pattern and the stromal tissues at the immediate vicinity of the tumor nests with strong heparanase expression were loose and edematous. Cystic areas and squamous metaplastic areas of the tumor showed intense staining with heparanase antibody proposing the implication of heparanase in these processes. These results suggest the possible contribution of heparanase in the local invasiveness and secondary morphologic changes of ameloblastoma. PMID- 15878762 TI - Effect of irrigated rice agriculture on Japanese encephalitis, including challenges and opportunities for integrated vector management. AB - Japanese encephalitis (JE) is a disease caused by an arbovirus that is spread by marsh birds, amplified by pigs, and mainly transmitted by the bite of infected Culex tritaeniorhynchus mosquitoes. The estimated annual incidence and mortality rates are 30,000--50,000 and 10,000, respectively, and the estimated global burden of JE in 2002 was 709,000 disability-adjusted life years lost. Here, we discuss the contextual determinants of JE, and systematically examine studies assessing the relationship between irrigated rice agriculture and clinical parameters of JE. Estimates of the sizes of the rural population and population in irrigated areas are presented, and trends of the rural population, the rice irrigated area, and the rice production are analyzed from 1963 to 2003. We find that approximately 1.9 billion people currently live in rural JE-prone areas of the world. Among them 220 million people live in proximity to rice-irrigation schemes. In 2003, the total rice harvested area of all JE-endemic countries (excluding the Russian Federation and Australia) was 1,345,000 km(2). This is an increase of 22% over the past 40 years. Meanwhile, the total rice production in these countries has risen from 226 millions of tonnes to 529 millions of tonnes (+134%). Finally, we evaluate the effect of different vector control interventions in rice fields, including environmental measures (i.e. alternate wet and dry irrigation (AWDI)), and biological control approaches (i.e. bacteria, nematodes, invertebrate predators, larvivorous fish, fungi and other natural products). We conclude that in JE-endemic rural settings, where vaccination rates are often low, an integrated vector management approach with AWDI and the use of larvivorous fish as its main components can reduce vector populations, and hence has the potential to reduce the transmission level and the burden of JE. PMID- 15878763 TI - Purification, cloning, and functional expression of phenylalanine aminomutase: the first committed step in Taxol side-chain biosynthesis. AB - The conversion of alpha-phenylalanine to beta-phenylalanine is the first committed step in the biosynthesis of the C-13 side chain of Taxol. Thus, the novel enzyme responsible for this step, phenylalanine aminomutase (PAM), is of considerable interest for studies of Taxol biosynthesis and represents a potential target for genetic engineering. A method is described for purifying PAM from Taxus chinensis cell cultures. The purified enzyme has a K(m) of 1.1mM, a V(max) of 110.1 microm/min/mg protein, a pH optimum of 7.5-8.0, and a denatured molecular weight of about 80 kDa. Peptide sequences derived from the purified protein were used to design and synthesize degenerate primers enabling the PCR synthesis of the PAM cDNA. The PAM cDNA encodes a protein of 687 amino acid residues with a deduced molecular weight of 75.3 kDa. The PAM cDNA was cloned and expressed in Escherichia coli, and PAM activity was demonstrated. As a gene symbol for the PAM enzyme, pam is proposed. Protein sequence alignments of PAM, phenylalanine ammonia-lyase (PAL), and histidine ammonia-lyase (HAL) sequences exhibit significant similarity providing insight into potential active site residues of PAM. PMID- 15878764 TI - Changes in the properties of mouse tongue muscle fibres before and after weaning. AB - The purpose of this study was to clarify any changes in muscle fibre properties in different regions of murine tongue during development, and to assess the effects of functional changes including weaning on these muscle fibres. The tongue was divided into upper and lower regions at the lateral margin, and the expression of myosin heavy chain (MHC) isoforms at different ages was investigated. Expression of genes encoding MHC proteins was quantified at the transcription level by quantitative reverse transcriptase polymerase chain reaction, and the protein expression of MHC isoforms was assessed by immunostaining. No difference was found in isoform expression between the upper and lower regions of the tongue before weaning. However, the expression of MHC-2b increased markedly in both regions after weaning, while that of MHC-2a decreased. At the age of 16 weeks, the expression of MHC-2b in the lower region was greater than that in the upper region. These findings show that during weaning, when there is a shift from sucking behaviour to mastication, the expression of MHC-2b increases along with an increase in the speed and strength of muscle contraction. Also, contraction force becomes stronger in the lower region of the tongue than the upper region at the age of 16 weeks. PMID- 15878765 TI - Regulatory effects of FGF-2 on the growth of mandibular condyles and femoral heads from newborn rats. AB - The secondary cartilage of the mandibular condyle is considered to be adaptive to functional factors. In the last decades, growth factors have also been shown to be potent regulators of cartilage metabolism. Moreover, it has been suggested that growth factors may differentially regulate the growth of primary and secondary cartilages. However, only a few studies have made a direct comparison of the effects of growth factors on both cartilages. Therefore, the aim here was to compare the effects of FGF-2 on secondary cartilage of the mandibular condyle and primary cartilage of the femoral head from 4-day-old rats in vitro. Cartilages were cultured for 1, 7 and 14 days with 0 and 100 ng/mL FGF-2. We evaluated the effects of FGF-2 on growth, tissue organisation, DNA and glycosaminoglycan (GAG) synthesis and GAG and collagen content. With FGF-2, the morphology of the mandibular condyles changed and the GAG and collagen contents were reduced. However, the growth of the mandibular condyles was not affected. On the contrary, the growth of the femoral heads was strongly reduced due to an inhibition of chondrocyte hypertrophy. In both cartilages, FGF-2 stimulated DNA synthesis in short-term cultures and reduced it in long-term cultures. In conclusion, FGF-2 had a larger effect on the metabolism of the mandibular condyles as compared to the femoral heads. However, the growth of the femoral heads was strongly reduced while that of the mandibular condyles was not affected. PMID- 15878766 TI - Tamoxifen alters gating of the BK alpha subunit and mediates enhanced interactions with the avian beta subunit. AB - Mammalian BK channels are modulated by estrogen and non-steroidal estrogen-like compounds (i.e. xenoestrogens), but the effects are dependent on channel composition. (Xeno)estrogens preferentially activate BK channels through accessory beta subunits, but reduce single-channel conductance by interaction with alpha subunits. In this report, the xenoestrogen tamoxifen was applied to chicken BK channels, in order to asses the mechanism behind drug interaction and to determine the extent to which (xeno)estrogen interaction is extended to avian BK homologs. As with mammalian isoforms, the properties of chicken BK channels were modulated by tamoxifen in a subunit-dependent manner. Tamoxifen reduced single-channel conductance through interaction with the alpha subunit. However, if the expression construct included the beta subunit, tamoxifen increased the channel's open probability and shifted the voltage-activation range to more negative potentials. This effect on channel gating was concentration-dependent, with an EC(50) of about 0.2 microM. Tamoxifen-mediated reductions in gating charge and in the intrinsic energetics that govern channel equilibrium. The relative contribution of these two effects on channel gating was altered by beta co-expression. Modulation by (xeno)estrogens may be an evolutionarily conserved mechanism for non-genomic hormonal actions, and the limited conservation between avian and mammalian beta subunits may suggest potential binding motifs. Alternatively, the data are consistent with a tamoxifen-mediated conformation change in the alpha subunit that alters the way alpha and beta subunits interact, resulting in enhanced gating without direct binding to beta. PMID- 15878767 TI - Antinociceptive tolerance to morphine from repeated nociceptive testing in the rat. AB - Repeated morphine administration has been shown to produce tolerance to the antinociceptive effects of morphine. However, the degree to which repeated morphine administration decreases antinociception is exaggerated by repeated behavioral testing, a phenomenon known as behavioral tolerance. An important question is whether behavioral tolerance can be overcome by direct administration of morphine into the ventrolateral periaqueductal gray (vPAG), a key structure contributing to morphine antinociception. Rats were injected with morphine or saline into the vPAG (Experiment 1) or subcutaneously (Experiment 2) followed 20 min later with hot-plate testing. The control groups received the same drug administration, but no nociceptive testing. Repeated nociceptive testing or repeated morphine administration produced antinociceptive tolerance regardless of whether morphine was injected into the vPAG or systemically. Administration of a high dose of morphine (20 mg/kg, s.c.) was able to overcome the development of behavioral tolerance, but not pharmacological tolerance revealing separate mechanisms for these two types of tolerance. These data indicate that behavioral tolerance is independent of the route of morphine administration. PMID- 15878768 TI - Effect of cholecalciferol supplementation on blood glucose in an experimental model of type 2 diabetes mellitus in spontaneously hypertensive rats and Wistar rats. AB - BACKGROUND: Vitamin D might have an influence on glucose concentrations, due to the presence of VDR receptors on the pancreas. We established an experimental model of type 2 diabetes in spontaneously hypertensive rats (SHR) and Wistar rats in order to investigate the glycemic response. METHODS: SHR males (n=6) and Wistar rats (n=6) weighing approximately 89+/-5.5 g and 123.5+/-6.5 g, respectively, after 7 days of basal period, had the chow pattern substituted (350 kcal/100 g) for a hypercaloric/hyperlipidic (HC/HL) diet (490 kcal/100g) and then injected with 40 mg/kg (SHR) and 20 mg/kg (Wistar) streptozotocin I.P. After the creation of diabetes, the rats suffered daily gavage of cholecalciferol (12.5 microg/kg(-) (1)) for 14 days. The blood glucose was assessed twice a week with a glucometer. The data were analyzed by ANOVA. RESULTS: SHR and Wistar rats fed on a HC/HL diet gained 60 g and 32 g in once week, vs. the basal period, where they only gained 23 g and 13 g, respectively. The cholecalciferol supplementation did not change the glucose concentration in all of the SHR animals. About 40% of the group responded by treatment with reduction of about 60% in glucose concentrations. We did find a 40% of the blood glucose levels in all Wistar rats. CONCLUSIONS: Cholecalciferol is able to reduce blood glucose in this experimental diabetes model. PMID- 15878769 TI - Mash1 is required for glomus cell formation in the mouse carotid body. AB - The carotid body consists of chemoreceptive glomus cells, sustentacular cells and nerve endings. The murine carotid body, located at the carotid bifurcation, is always joined to the superior cervical ganglion of the sympathetic trunk. Glomus cells and sympathetic neurons are immunoreactive for the TuJ1, PGP9.5, tyrosine hydroxylase (TH) and neuropeptide Y (NPY) markers. Glomus cells are also immunoreactive for serotonin (5-HT). A targeted mutation of Mash1, a mouse homolog of the Drosophila achaete-scute complex, results in the elimination of sympathetic ganglia. In Mash1 null mutant mice, the carotid body primordium forms normally in the wall of the third arch artery at embryonic day (E) 13.0 and continues to develop, although the superior cervical ganglion is completely absent. However, no cells in the mutant carotid body display the TuJ1, PGP 9.5, TH, NPY and 5-HT markers throughout development. The absence of glomus cells was also confirmed by electron microscopy. The carotid body of newborn null mutants is composed of mesenchymal-like cells and nerve fibers. Many cells immunoreactive for the S-100 protein, a sustentacular cell marker, appear in the mutant carotid body during fetal development. The Mash1 gene is thus required for the genesis of glomus cells but not for sustentacular cells. PMID- 15878770 TI - Partial protection against Eimeria acervulina and Eimeria tenella induced by synthetic peptide vaccine. AB - Coccidiosis is a major parasitic disease of poultry industry and an ideal vaccine should induce long-lasting cross-species protective immunity. Broiler chickens (Cobb 500) were inoculated with single, double or triple injections of a synthetic peptide (derived from sequences of Eimeria acervulina and Eimeria tenella antigens) homogenized in Freund's complete and incomplete adjuvants. The immune responses to the vaccine were assessed by evaluation of antibody and lymphocyte proliferation responses, and the degree of resistance of vaccinated chickens to challenge with sporulated oocysts of E. acervulina or E. tenella determined by comparison of their oocyst output with those of control chickens. The results indicated that the synthetic peptide vaccine induced a high level of antibody and cellular responses associated with partial cross-species protection against challenge with sporulated oocysts of E. acervulina or E. tenella. PMID- 15878771 TI - Nurse turnover: a literature review. AB - Ongoing instability in the nursing workforce is raising questions globally about the issue of nurse turnover. A comprehensive literature review was undertaken to examine the current state of knowledge about the scope of the nurse turnover problem, definitions of turnover, factors considered to be determinants of nurse turnover, turnover costs and the impact of turnover on patient, and nurse and system outcomes. Much of the research to date has focused on turnover determinants, and recent studies have provided cost estimations at the organizational level. Further research is needed to examine the impact of turnover on health system cost, and how nurse turnover influences patient and nurse outcomes. PMID- 15878772 TI - The recovery of dysfunctional lipoprotein lipase (Asp204-Glu) activity by modification of substrate. AB - Functional deficiency of lipoprotein lipase (LPL) was found in a patient with severe hypertriglyceridemia. The patient was 39-year-old man with a plasma triglyceride level of 2032 mg/dl, and suffered from recurrent pancreatitis. His post heparin plasma LPL mass was almost normal, but the LPL activity was remarkably decreased. Gene analysis showed that homozygote missense mutation (204 Asp (GAC)-Glu (GAG)) exists in exon 5 of LPL gene. The patient LPL purified from post heparin plasma scarcely hydrolyzed VLDL-triglyceride and also triolein emulsified with Triton X-100 or phosphatidylcholine. When phosphatidylethenolamine, phosphatidylserine and cardiolipin were used as an emulsifier for triolein, triolein-hydrolyzing activity of the patient's LPL was observed and was much higher than that of wild-type LPL. Mutant LPL gene (Asp204 Glu) was made by site-direct mutagenesis and was transfected to COS-1 cell. The expressed LPL (Asp204-Glu) also showed the same properties. These results suggested that the LPL (Asp204-Glu) is a functional deficiency, and the activity could be recovered by using acidic phospholipids as an emulsifier. PMID- 15878773 TI - An Endoreticulatus species from Ocinara lida (Lepidoptera: Bombycidae) in Taiwan. AB - A microsporidium from the Ficus pest, Ocinara lida, in Taiwan is characterized. The taxonomic position of this species was preliminarily determined by sequencing small subunit rRNA gene (SSUrRNA). Analysis of the SSUrRNA sequence indicated that this isolate from O. lida is a member of the genus Endoreticulatus and belongs to the genetic grouping containing other lepidopteran Endoreticulatus species we have analyzed phylogenetically. The taxonomic position of this isolate was also confirmed by the ultrastructural characteristics of this isolate. The congruence between SSUrRNA sequence analysis and ultrastructural characteristics shows that this isolate is more closely related to Endoreticulatus bombycis than to Endoreticulatus schubergi Zwolfer. PMID- 15878774 TI - Management commitment to safety as organizational support: relationships with non safety outcomes in wood manufacturing employees. AB - INTRODUCTION: Employee perceptions of management commitment to safety are known to influence important safety-related outcomes. However, little work has been conducted to explore non-safety-related outcomes resulting from a commitment to safety. METHOD: Employee-level outcomes critical to the effective functioning of an organization, including attitudes such as job satisfaction and commitment to the organization, were included on surveys given to 641 hourly production employees at three wood products manufacturing facilities. Participants' were asked about perceptions of management commitment to safety and job-related variables such as perceived dangerousness of their position, organizational commitment, and withdrawal behaviors. Supervisors also rated the performance of each of their hourly subordinates. RESULTS: Results suggest that employee outcomes differ based on perceptions of management's commitment to safety. Specifically, management commitment to safety was positively related to job satisfaction, organizational commitment, and job-related performance. We also found a negative relationship between commitment to safety and employee withdrawal behaviors. CONCLUSIONS: Our results suggest that increasing employee perceptions of management's personal concern for employee well-being through a dedication to safety will result in positive outcomes beyond improved safety performance. These results also imply that there is a type of social exchange between employees and management that may affect employees similarly to perceived organizational support. IMPACT ON INDUSTRY: Results further reinforce the value of a commitment to safety by a firm's management. Organizations with a strong commitment to safety may enjoy not only a reduction in safety-related events but also increases in desirable employee attitudes and behaviors. PMID- 15878775 TI - Work domain analysis for air traffic controller weather displays. AB - INTRODUCTION: Adverse weather conditions have a major impact on National Airspace System (NAS) operations. They create safety hazards for pilots, constrain the usable airspace for air traffic control (ATC), and reduce the overall capacity of the NAS. A system-wide dissemination of weather information to controllers could theoretically improve safety and efficiency. PROBLEM: However, it is currently unclear what weather information would be beneficial for tactical operations. Furthermore, no previous research has empirically evaluated optimal presentation designs for ATC weather displays. Ill-designed weather displays can cause safety hazards by presenting redundant information (i.e., by increasing the cognitive load) and display clutter (e.g., by interfering with the visual extraction of traffic data). METHOD: In the present paper, we outline our use of cognitive work analysis (CWA) techniques for the assessment of weather information needs for terminal controllers. RESULTS: Specifically, we describe how the CWA modeling tools helped us reveal instances in the terminal domain where weather information is lacking or insufficiently disseminated. We used our CWA results to drive the development of weather display concepts and to set up a high-fidelity simulation capability. IMPACT ON INDUSTRY: By means of high-fidelity simulations, we can empirically evaluate controller weather information needs in order to propose weather displays for increased aircraft safety and efficiency of terminal operations. PMID- 15878776 TI - Parental risk perceptions of childhood pedestrian road safety: a cross cultural comparison. AB - INTRODUCTION: This cross-sectional community-based randomized telephone survey, aimed to compare the parental risk perceptions on childhood pedestrian road safety among three different major community language speaking groups and the mainstream English speaking Australians in Sydney, Australia. METHOD: This was a community-based telephone survey with subjects randomly selected from the population of specific cultural languages in Sydney. RESULTS: Results indicated that Chinese and Arabic speaking parents or caregivers perceived the road environments as significantly less hazardous for their 4-12 year old children as pedestrians when compared with Vietnamese and English speaking parents. This was true even after adjusting for the age of child, socioeconomic status, and the living environments of respondents. CONCLUSION: The implication of the results was discussed in light of possible application to childhood safety behavior across different cultures. IMPACT TO THE INDUSTRY: The results obtained have highlighted the importance of cultural factors in risk perceptions, and safety behaviors. Cultural differences in risk perceptions should be taken into consideration in the design of safety education programs. PMID- 15878777 TI - Toxicogenomic analysis of gene expression changes in rat liver after a 28-day oral benzene exposure. AB - Benzene is an industrial chemical, component of automobile exhaust and cigarette smoke. After hepatic bioactivation benzene induces bone marrow, blood and hepatic toxicity. Using a toxicogenomics approach this study analysed the effects of benzene at three dose levels on gene expression in the liver after 28 daily doses. NMR based metabolomics was used to assess benzene exposure by identification of characteristic benzene metabolite profiles in urine. The 28-day oral exposure to 200 and 800 mg/kg/day but not 10 mg/kg/day benzene-induced hematotoxicity in male Fisher rats. Additionally these upper dose levels slightly reduced body weight and increased relative liver weights. Changes in hepatic gene expression were identified with oligonucleotide microarrays at all dose levels including the 10 mg/kg/day dose level where no toxicity was detected by other methods. The benzene-induced gene expression changes were related to pathways of biotransformation, glutathione synthesis, fatty acid and cholesterol metabolism and others. Some of the effects on gene expression observed here have previously been observed after induction of acute hepatic necrosis with bromobenzene and acetaminophen. In conclusion, changes in hepatic gene expression were found after treatment with benzene both at the toxic and non-toxic doses. The results from this study show that toxicogenomics identified hepatic effects of benzene exposure possibly related to toxicity. The findings aid to interpret the relevance of hepatic gene expression changes in response to exposure to xenobiotics. In addition, the results have the potential to inform on the mechanisms of response to benzene exposure. PMID- 15878778 TI - INK4a/ARF: a multifunctional tumor suppressor locus. AB - The INK4a/ARF locus encodes two physically linked tumor suppressor proteins, p16(INK4a) and ARF, which regulate the RB and p53 pathways, respectively. The unusual genomic relationship of the open reading frames of these proteins initially fueled speculation that only one of the two was the true tumor suppressor, and loss of the other merely coincidental in cancer. Recent human and mouse genetic data, however, have firmly established that both proteins possess significant in vivo tumor suppressor activity, although there appear to be species- and cell-type specific differences between the two. For example, ARF plays a clear role in preventing Myc-induced lymphomagenesis in mice, whereas the role for p16(INK4a) is human carcinomas is more firmly established. In this review, I discuss the evolutionary history of the locus, the relative importance of these tumor suppressor genes in human cancer, and recent information suggesting novel biochemical and physiologic functions of these proteins in vivo. PMID- 15878779 TI - Endocannabinoid release from midbrain dopamine neurons: a potential substrate for cannabinoid receptor antagonist treatment of addiction. AB - Substantial evidence suggests that all commonly abused drugs act upon the brain reward circuitry to ultimately increase extracellular concentrations of the neurotransmitter dopamine in the nucleus accumbens and other forebrain areas. Many drugs of abuse appear to increase dopamine levels by dramatically increase the firing and bursting rates of dopamine neurons located in the ventral mesencephalon. Recent clinical evidence in humans and behavioral evidence in animals indicate that cannabinoid receptor antagonists such as SR141716A (Rimonabant) can reduce the self-administration of, and craving for, several commonly addictive drugs. However, the mechanism of this potentially beneficial effect has not yet been identified. We propose, on the basis of recent studies in our laboratory and others, that these antagonists may act by blocking the effects of endogenously released cannabinoid molecules (endocannabinoids) that are released in an activity- and calcium-dependent manner from mesencephalic dopamine neurons. It is hypothesized that, through the antagonism of cannabinoid CB1 receptors located on inhibitory and excitatory axon terminals targeting the midbrain dopamine neurons, the effects of the endocannabinoids are occluded. The data from these studies therefore suggest that the endocannabinoid system and the CB1 receptors located in the ventral mesencephalon may play an important role in regulating drug reward processes, and that this substrate is recruited whenever dopamine neuron activity is increased. PMID- 15878780 TI - Genomics refutes an exclusively African origin of humans. AB - Ten years ago, evidence from genetics gave strong support to the "recent African origin" view of the evolution of modern humans, which posits that Homo sapiens arose as a new species in Africa and subsequently spread, leading to the extinction of other archaic human species. Subsequent data from the nuclear genome not only fail to support this model, they do not support any simple model of human demographic history. In this paper, we study a process in which the modern human phenotype originates in Africa and then advances across the world by local demic diffusion, hybridization, and natural selection. While the multiregional model of human origins posits a number of independent single locus selective sweeps, and the "out of Africa" model posits a sweep of a new species, we study the intermediate case of a phenotypic sweep. Numerical simulations of this process replicate many of the seemingly contradictory features of the genetic data, and suggest that as much as 80% of nuclear loci have assimilated genetic material from non-African archaic humans. PMID- 15878781 TI - Sb(III) oxidation by iodate in seawater: a cautionary tale. AB - Knowledge of antimony redox kinetics is crucial in understanding the impact and fate of antimony in the environment. The oxidation of Sb(III) with iodate was measured in 0.5 mol L(-1) NaCl solutions as a function of pH at environmentally significant concentrations of antimony and iodate. The oxidation of Sb(III) with iodate is pH dependent: no measurable oxidation is observed below pH 9. The undissociated Sb(OH)3 does not react with iodate and the formation of significant amounts of Sb(OH)4- is needed for the reaction to take place. It is thus unlikely that iodate oxidizes Sb(III) in seawater. Our results support that the observed presence of the thermodynamically unstable Sb(III) in oxic waters can be due to the kinetic stabilization of the trivalent state vis-a-vis some common abiotic oxidants at natural pH values. However, caution must be exercised because the presence of iodate in seawater favours fast oxidation of Sb(III) if water samples are acidified, as is the case in many analytical procedures. PMID- 15878782 TI - Method to compare mu-tensile bond strength of a self-etching adhesive and mu cohesive strength of adjacent dentin. AB - Laboratory results from tensile or micro-tensile testing of adhesive/restorative systems need a gold standard to make their interpretation possible. This can be done by comparing the mu-tensile bond strength (muTBS) of the tested adhesive(s) with the mu-tensile cohesive strength (muTCS) of the adjacent dentin, which is meant to be replaced by the restoration. OBJECTIVES: To test immediate muTBS of an adhesive/restorative system versus muTCS of the dentin adjacent to the bonded interface, in perfused specimens. METHODS: Enamel and roots of five-third sound molars were removed and teeth were perfused (30 cm distilled H2O) until after the bonding procedure was completed. Parallel grooves (1.5-mm widex 1.5-mm deep, separated by 1.5mm), were drilled in the exposed dentin, under water refrigeration. Teeth were restored (AdheSE/Filtek Z250). Specimens were sawn from grooves to test muTBS (n = 30) and from between grooves to test muTCS (n = 32) of dentin, adjacent to adhesive interfaces, both areas to be tested at the same relative depth. Areas to be studied were rounded (mean bonded surface area for both groups, BA = 0.68 mm2), and muTBS and muTCS were found. As not all BAs were identical, residuals of muTBS and muTCS values to the regression line relating BA and tensile test results (representing the null hypothesis, i.e. muTBS = muTCS) were compared using a non-parametric test. RESULTS: Difference in the means of residuals was not statistically significant (two tails p = 0.067). SIGNIFICANCE: mean muTBS of the tested adhesive was not different to mean muTCS of adjacent dentine. The null hypothesis was not rejected. PMID- 15878783 TI - Electrochemical impedance spectroscopy study of Ti-6Al-4V alloy in artificial saliva with fluoride and/or bovine albumin. AB - OBJECTIVES: To investigate the electrochemical mechanism of Ti-6Al-4V alloy in acidic artificial saliva with NaF and/or bovine albumin (BA). METHODS: The electrochemical behavior of Ti-6Al-4V alloy was investigated using electrochemical impedance spectroscopy (EIS) measurements at the open circuit potentials. The electrolytes used included artificial saliva with 0-0.5% NaF, with 0.1% NaF+0.01-0.5% BA, and with 0.01-0.5% BA at pH 5 and 37 degrees C. One way ANOVA was used to analyze the influence of NaF and BA concentration on the polarization resistance (Rp) obtained from the EIS data. Surface chemical analyses were performed with X-ray photoelectron spectroscopy. RESULTS: The impedance was mainly characterized by the resistance and capacitance of a protective TiO2-based passive film on Ti-6Al-4V alloy when NaF concentration was low (< or = 0.01%). When NaF concentration was higher than 0.1%, the impedance was characterized by two capacitive and one inductive effects, which were related to the metal's charge transfer reaction and the decayed TiO2-based oxide film (capacitive effects), and the relaxation of the adsorbed Ti-F complex compound (inductive effect). The addition of 0.01-0.5% BA in 0.1% NaF-containing artificial saliva significantly increased the impedance of Ti-6Al-4V alloy compared to that without BA addition (p < 0.001). The presence of BA in fluoride free artificial saliva had no influence on the electrochemical impedance of Ti 6Al-4V alloy (p > 0.05). SIGNIFICANCE: The EIS measurement technique is suitable for the study of the electrochemical behavior of dental alloy. The electrochemical mechanism of Ti-6Al-4V alloy in artificial saliva is related to the fluoride and bovine albumin concentration. PMID- 15878784 TI - The influence of calcium fluoride (CaF2) on biaxial flexural strength of apatite mullite glass-ceramic materials. AB - OBJECTIVE: The aim of this study was to evaluate the effect of varying the molar percentage of calcium fluoride (CaF2) on the biaxial flexural strength (BFS) of apatite-mullite glass-ceramics. METHODS: Three apatite-mullite glass-ceramic materials were produced based on a formula of 4.5SiO(2)-3Al2O(3)-1.5P2O(5)-3CaO XCaF2, where X=0.5, 1 and 3 and called HG 1-3, respectively. These materials were used to produce discs of 2 mm thickness and 12 mm diameter using the lost-wax casting process. 30 discs per material were produced, 10 discs per material were cast and then left to bench cool (glass state), 10 discs per material were given a heat treatment at 765, 679 and 629 degrees C for 1h and then heat treated at 890, 860 and 824 degrees C for the HG 1-3, respectively, for another hour to form apatite. Ten discs per material were heat treated as previously described (765, 679 and 629 degrees C) then heat treated for 1h at 1022, 987 and 892 degrees C for the HG 1-3, respectively, to form apatite-mullite. The heat treatment temperatures were obtained from differential thermal analysis data. A lithium disilicate glass-ceramic was used as a control. Biaxial flexural strength (BFS) was determined using a LIoyd 2000R tester. RESULTS: Data showed that the BFS increased as the fluoride content increased, and the apatite-mullite samples had significantly higher BFS values than the as cast glass or apatite samples (p<0.05), with the control having significantly higher BFS values than all the HG glass-ceramic materials for every condition (p<0.05). The fictive glass transition temperature (Tg) was observed to drop with increasing fluoride content. SIGNIFICANCE: Increasing the CaF2 content increased the BFS and decreased the Tg of the glass-ceramic materials tested. PMID- 15878785 TI - Suppression of glial activation is involved in the protection of IL-10 on maternal E. coli induced neonatal white matter injury. AB - White matter damage (WMD) is an important cause of disability including cerebral palsy in preterm, low birth-weight infants. Maternal infection is now recognized as one of the risk factors for WMD. Previously we reported that intrauterine inoculation of Escherichia coli to pregnant rats resulted in WMD in offspring and interleukin-10 (IL-10) was protective against this damage. The objective of this study was to elucidate the mechanism involved in the protective effect of IL-10 against neonatal WMD. We found that E. coli treatment in dams resulted in significant apoptosis in periventricular white matter of rat pups on postnatal day 0 (P0). On P8, a remarkable increase in ED-1 immunostaining (indicating either microglial activation or macrophage infiltration) was detected in brains of pups in the E. coli-treated group. Astrogliosis was also noticed in brain white matter of pups in the E. coli-treated group. In addition to the strong activation of microglia and astrocytes, oligodendrocytes (OLs) were significantly reduced in periventricular areas in the brains of pups from the E. coli-treated group. Later, on P15, hypomyelination was also noticed in rat brains from the E. coli-treated group, using myelin basic protein (MBP) immunostaining. Treatment with IL-10 after E. coli inoculation significantly reduced TUNEL staining and caspase-3 activation, and partially restored the impaired immunostaining markers for immature and mature OLs, such as CNPase, O4, adenomatous polyposis coli (APC) and MBP. These results indicate that the protective effect of IL-10 against brain WMD is linked with suppression of microglial activation/macrophage infiltration, as shown by significantly reduced ED-1+ cells in the white matter. PMID- 15878786 TI - Development of antiviral therapy for severe acute respiratory syndrome. AB - A new disease, the severe acute respiratory distress syndrome (SARS), caused by the SARS coronavirus (SARS-CoV), emerged at the beginning of 2003 and rapidly spread throughout the world. Although the disease had disappeared in June 2003 its re-emergence cannot be excluded. The development of vaccines against SARS-CoV may take years. Therefore, the availability of effective antiviral drugs against SARS-CoV may be crucial for the control of future SARS outbreaks. In this review, experimental and clinical data about potential anti-SARS drugs is summarised and discussed. Animal model studies will be needed to help to determine which interventions warrant controlled clinical testing. PMID- 15878787 TI - Effects of different malnutrition techniques on the behavior of rats tested in the elevated T-maze. AB - The influence of different malnutrition techniques on the behavior of adult animals was investigated in the elevated T-maze (ETM). Control litters (C) were composed by eight pups constantly kept with their mother and fed by a 16%-protein diet ad libitum; protein malnutrition litters (PM) were fed by a 6%-protein diet; protein-calorie malnutrition litters (PCM) were fed with 50% of the 16%-protein diet ingested by C litters; malnutrition by increase in the size of the litter (LLM-number of pups was twice the number of pups in C litters), and malnutrition by separation (SM-litters spent half of the day with non-lactating females). After weaning, all groups received lab chow diet until the test day (70th day). During the test were recorded the basal, avoidance 1, avoidance 2 and escape latencies. The data showed that PM, PCM, LLM and SM animals showed lower increases in avoidance latencies, when compared to their control groups. However, malnutrition did not affect escape latencies. The nature of these alterations seems to be nutritional, as the extra-nutritional factors (i.e. maternal care) differ a lot among the malnutrition techniques. These results suggest that malnutrition, irrespective of the technique, altered the neural mechanisms believed to control defensive behaviors in the ETM. PMID- 15878788 TI - Effect of major ions on the toxicity of copper to Hyalella azteca and implications for the biotic ligand model. AB - The effect of major ions (Ca, Mg, Na, and K) and pH on Cu toxicity (LC50) to Hyalella azteca was determined in 1 week exposures. The simplest equation for describing Cu toxicity is a linear relationship between the total dissolved Cu LC50 and Ca and Na in water, ignoring pH. This equation would be useful in tier one of a two-tiered approach; if the measured dissolved Cu exceeds the value predicted from the equation, the sample should either be tested for toxicity, or a more detailed chemical speciation analysis can be conducted. The data were not consistent with a single-binding-site biotic ligand model, assuming that toxicity was due to the free Cu ion alone. However, toxicity could be predicted using a two-binding-site model. This requires separate coefficients to account for the effects of Ca and Na at low and high pH values (6.5-8.4), corresponding to the different binding sites (Mg and K did not affect toxicity). The single-binding site BLM does not allow for this. Toxicity of Cu hydroxide or carbonate complexes does not need to be invoked, but cannot be excluded, and several models invoking the toxicity of these complexes can also explain the data. The free ion LC50 is strongly dependent on pH, but the LC50 for total dissolved Cu is almost pH independent. The effects of Ca and Na on the free ion LC50 are very different at high and low pH (contrary to single-site biotic ligand model predictions), but similar for total dissolved Cu. Published data suggest that the same model, with different coefficients, can also be applied to Daphnia and fish. A more critical evaluation of the effects of cations at both low and high pH for organisms other than Hyalella is needed to determine if the BLM needs to be adjusted to incorporate more than one binding site for other species as well. Hydrogen ions reduce the toxicity of free Cu ions to Hyalella, but Cu also reduces the toxicity of hydrogen ions. A mixture model accounting for the joint toxicity of Cu and pH, as well as their mutual antagonistic effects, is presented. PMID- 15878789 TI - Gene polymorphism of Plasmodium falciparum merozoite surface proteins 4 and 5. PMID- 15878790 TI - Biochemical and genetic analysis of the distinct proliferating cell nuclear antigens of Toxoplasma gondii. AB - The apicomplexa parasite Toxoplasma gondii expresses two distinct proliferating cell nuclear antigens (PCNA) that exhibit distinct patterns of subcellular localization during tachyzoite growth. In all cell cycle phases, TgPCNA1 is concentrated in the nucleus, while TgPCNA2 is only concentrated in the nucleus during S-phase and uniformly distributed throughout the cell during mitosis and early G1-phase. TgPCNA1-GFP and native TgPCNA2 display a punctate staining pattern that is consistent with assembly into replication foci during S-phase; however, TgPCNA2 disassociates from replication foci before TgPCNA1-GFP. Consistent with the distinct pattern of TgPCNA2 cellular localization, homotypic TgPCNA2 interactions were primarily observed by yeast two-hybrid or co immunoprecipitation analysis. Transgenic parasites in which the TgPCNA2 gene was disrupted displayed a slower growth rate in vitro; however, no difference in DNA polymerase activity, response to chemical mutagens, or recombinational frequency was observed in these mutant clones demonstrating that TgPCNA2 is non-essential in the tachyzoite developmental stage. Heterologous expression of TgPCNA1, but not TgPCNA2, was able to complement a POL30 cold-sensitive yeast strain suggesting that this isoform may serve as a major replisomal factor in T. gondii and is consistent with the failure to disrupt this gene in tachyzoites. PMID- 15878791 TI - Recombinant non-structural 1 (NS1) protein of dengue-2 virus interacts with human STAT3beta protein. AB - A combination of yeast two-hybrid library screening, co-immunoprecipitation and immunofluorescence microscopy demonstrated that dengue-2 virus non-structural 1 (NS1) protein can interact with an N-terminally truncated form of human STAT3beta (DeltaN40-STAT3beta) protein. The NS1 protein interacted with the activated STAT3beta protein in vesicle-like structures in the cell cytoplasm. In addition, transfection of dendritic cells with plasmid expressing NS1 protein also resulted in significant induction of tumor necrosis factor-alpha (TNFalpha) and interleukin-6 (IL-6). Since the STAT3beta protein is an acute-phase response factor, its interaction with NS1 protein may influence the pathological changes observed in dengue fever, dengue hemorrhagic fever and dengue shock syndrome. PMID- 15878792 TI - Substantial increases in idarubicin plasma concentration by liposome encapsulation mediates improved antitumor activity. AB - Idarubicin has been successfully encapsulated in cholesterol-free liposomes, however, little is known about how the rate of drug release from circulating liposomes influences therapeutic activity. The studies described herein assess the attributes of a liposome formulation required to significantly increase the plasma levels of idarubicin and further establish whether increases in the circulation longevity of the drug mediate improved antitumor activity. Pharmacokinetic assessments of 6 different 3[H]-labelled liposome formulations were compared to free idarubicin. The highest idarubicin plasma concentrations were observed with DSPC/DSPE-PEG2000 liposomes formulated with 2 mol% DSPE PEG2000 and 150 mM (iso-osmotic) internal citrate concentration. It was shown that increased levels of PEG-lipid incorporation augmented IDA release and the optimal liposomal formulation needed to be prepared under iso-osmotic conditions. For efficacy studies in a murine leukemia model, groups of 12-14 mice were treated i.v. with saline or equivalent doses (1, 2, 3 mg/kg) of free or liposomal IDA. Liposomal treatment groups exhibited a higher % increase in life span (ILS) as compared to equivalent doses of free drug. Efficacy studies completed in two drug resistant models, P388/ADR and MDA435LCC6/MDR1, demonstrated that neither the free nor liposomal formulation of idarubicin was therapeutically active. Encapsulation of IDA in liposomes increased antitumor activity in an IDA sensitive model, however, the significant increase in plasma drug levels was not sufficient to overcome multidrug resistance. PMID- 15878793 TI - Effects of alpha-lipoic acid on kinin B1 and B2 receptor binding sites in the spinal cord of chronically angiotensin-treated rats. AB - A quantitative autoradiographic study was performed to determine whether kinin receptors are altered in the rat spinal cord in an experimental model of arterial hypertension under antioxidant therapy with alpha-lipoic acid. Sprague-Dawley rats were fed for 4 weeks with a normal chow diet or with an alpha-lipoic acid supplemented diet (1000 mg/kg feed), and treated for the last 2 weeks with angiotensin II (AT II) (200 ng/kg/min with an osmotic pump implanted s.c.). Control rats received either diet but not AT II. A 2-week administration of AT II increased significantly systolic blood pressure, the production of superoxide anion in the aorta and B1 receptor binding sites in the thoracic spinal dorsal horn. This treatment did not affect spinal B2 receptor binding sites, glycemia and insulinemia. The diet supplemented with alpha-lipoic acid reduced significantly the increase in systolic blood pressure, the production of aortic superoxide anion and prevented the increases of B1 receptor binding sites. Results show an association between the oxidative stress and the increases of B1 receptors and arterial blood pressure induced by AT II. Data also exclude the possibility that arterial hypertension is a primary mechanism leading to an increase of B2 receptor binding sites in the rat spinal cord. PMID- 15878794 TI - Treatment of severe pulmonary hypertension: a bradykinin receptor 2 agonist B9972 causes reduction of pulmonary artery pressure and right ventricular hypertrophy. AB - Bradykinin is an important modulator of endothelial cell function and has also a powerful cardioprotective effect. Here we report that treatment of severely pulmonary hypertensive rats (that recapitulate several of the physiological and pathological characteristics of the human pulmonary vascular disease, including dramatic right ventricular hypertrophy, pericardial effusion and death) with a newly synthesized long-acting bradykinin B2 receptor agonist B9972 caused reduction of the pulmonary artery pressure (PAP=51+/-2.0 versus 68+/-2.8 of untreated animals) and of right ventricular hypertrophy (Rv/Lv+S=0.55+/-0.02 versus 0.73+/-0.03 of untreated rats) and activation of Akt. Long-term stimulation with B9972 in our animal model of SPH resulted in decreased expression of the B2 receptor in lung vasculature. Treatment with B9972 decreased the number of plexiform lesions in the lungs by inducing cell apoptosis in the obliterated vessels and by restoring caveolin-1 expression. B9972 also promoted eNOS activation. In vitro B9972 caused activation of caspase-3 as well as Erk and induction of prostacyclin production in rat pulmonary microvascular EC. Taken together our data suggest that a stable bradykinin B2 agonist B9972 demonstrates the capacity to reduce severe pulmonary hypertension, right ventricular hypertrophy and induce apoptosis of hyperproliferative cells in pre-capillary pulmonary arterioles. PMID- 15878795 TI - Combination cancer chemotherapy with one compound: pluripotent bradykinin antagonists. AB - Lung and prostate cancers are major health problems worldwide. Treatments with standard chemotherapy agents are relatively ineffective. Combination chemotherapy gives better treatment than a single agent because the drugs can inhibit the cancer in different pathways, but new therapeutic agents are needed for the treatment of both tumor types. Bradykinin (BK) antagonists offer advantages of combination therapy in one compound. These promising multitargeted anti-cancer compounds selectively stimulate apoptosis in cancers and also inhibit both angiogenesis and matrix metalloprotease (MMP) action in treated lung and prostate tumors in nude mice. The highly potent, metabolism-resistant bradykinin antagonist peptide dimer, B-9870 [SUIM-(DArg-Arg-Pro-Hyp-Gly-Igl-Ser-DIgl-Oic Arg)2] (SUIM=suberimidyl; Hyp=4-hydroxyproline; Igl=alpha-(2-indanyl)glycine; Oic=octahydroindole-2-carboxylic acid) and its non-peptide mimetic, BKM-570 [2,3,4,5,6-pentafluorocinnamoyl-(o-2,6-dichlorobenzyl)-L-tyrosine-N-(4-amino 2,2,6,6-tetramethylpiperidyl)amide] are superior to the widely used but toxic chemotherapeutic drugs cisplatin and taxotere. In certain combinations, they act synergistically with standard anti-cancer drugs. Due to its structure and biological activity, BKM-570 is an attractive lead compound for derivatization and evaluation for lung and prostate cancer drugs. PMID- 15878796 TI - Influence of glyphosate and its formulation (Roundup) on the toxicity and bioavailability of metals to Ceriodaphnia dubia. AB - This study examined the toxicological interaction between glyphosate (or its formulation, Roundup) and several heavy metals to a freshwater cladoceran, Ceriodaphnia dubia. We demonstrated that all binary combinations of Roundup and metals (Cd, Cu, Cr, Ni, Pb, Se and Zn) exhibited "less than additive" mixture toxicity, with 48-h LC50 toxic unit > 1. Addition of glyphosate alone could significantly reduce the acute toxicity of Ag, Cd, Cr, Cu, Ni, Pb and Zn (but not Hg and Se). The ratio between glyphosate and metal ions was important in determining the mitigation of metal toxicity by glyphosate. A bioaccumulation study showed that in the presence of glyphosate the uptake of some metals (e.g. Ag) was halted but that of others (e.g. Hg) was increased significantly. Therefore, our study strongly suggests that glyphosate and its commercial formulations can control the toxicity as well as the bioavailability of heavy metals in aquatic ecosystems where both groups of chemicals can co-occur. PMID- 15878797 TI - Purple grape juice inhibits 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumorigenesis and in vivo DMBA-DNA adduct formation. AB - There has been considerable interest in identifying specific foods and phytochemicals that may have breast cancer preventive properties. Concord grapes are rich in polyphenolic chemicals and anthocyanin pigments that may have biological properties which could suppress cancer such as having antioxidant, antiproliferative, and proapoptotic actions. To determine the potential breast cancer protective action of purple grape juice, we examined the effect of grape juice consumption on the initiation stage of 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumorigenesis and on the in vivo formation of rat mammary DNA adducts in female Sprague-Dawley rats. Consumption of grape juice significantly inhibited mammary tumor mass at termination and the growth of tumors for the first 5 weeks of detectable tumor development. Consumption of grape juice phenolics by rats also significantly inhibited in vivo mammary DMBA DNA adduct formation by 34 and 56% for animals fed phenolics at 346 and 692 mg/dL, respectively, compared to controls. Mammary 8-oxo-deoxyguanosine (8-oxo dG) levels decreased by 25 and 37%, respectively, but the differences were not statistically significant. Liver DMBA-DNA adducts decreased by 10-30%, while 8 oxo-dG adducts remained unchanged, following grape juice intake. Liver glutathione S-transferase activity was significantly increased following grape juice consumption, but only at the highest level of intake. In addition, liver activities of catalase increased and xanthine oxidase decreased significantly, but only at the highest grape juice dose. Thus, these studies indicate that specific constituents or combinations of phytochemicals in purple grape juice can block the initiation stage of DMBA-induced rat mammary tumorigenesis. This tumor inhibitory effect was associated with a suppression of mammary DMBA-DNA adduct formation, which in part may be explained by increased liver activity of the phase II metabolizing enzyme, glutathione S-transferase. Mammary and liver 8-oxo dG levels were not significantly altered by grape juice consumption. Thus, grape juice constituents appear to have benefit in decreasing susceptibility of the rat mammary gland to the tumor-initiating action of DMBA. PMID- 15878798 TI - Removal of heavy metal ions from aqueous solutions using carbon aerogel as an adsorbent. AB - The removal of Cd(II), Pb(II), Hg(II), Cu(II), Ni(II), Mn(II) and Zn(II) by carbon aerogel has been found to be concentration, pH, contact time, adsorbent dose and temperature dependent. The adsorption parameters were determined using both Langmuir and Freundlich isotherm models. Surface complexation and ion exchange are the major removal mechanisms involved. The adsorption isotherm studies clearly indicated that the adsorptive behaviour of metal ions on carbon aerogel satisfies not only the Langmuir assumptions but also the Freundlich assumptions, i.e. multilayer formation on the surface of the adsorbent with an exponential distribution of site energy. The applicability of the Lagergren kinetic model has also been investigated. Thermodynamic constant (K(ad)), standard free energy (DeltaG(0)), enthalpy (DeltaH(0)) and entropy (DeltaS(0)) were calculated for predicting the nature of adsorption. The results indicate the potential application of this method for effluent treatment in industries and also provide strong evidence to support the adsorption mechanism proposed. PMID- 15878799 TI - The effects of regular aerobic exercise in adolescent period on hippocampal neuron density, apoptosis and spatial memory. AB - It is known that positive effects of regular aerobic exercise on cognitive functions in humans and also animals; but how to the effects of aerobic exercise in adolescent period is unknown. The present study examined the effects of regular aerobic exercise on spatial memory using the Morris water maze, cell density and apoptosis of hippocampus in adolescent rats. Twenty-two days of age male rats were run on a treadmill for 30 min/session at a speed of 8m/min and 0 degrees slope, five times a week for 8 weeks. The present study showed that exercise induced significant cognitive improvement throughout brain maturation in rats. The number of hippocampal CA1 and CA3 neurons, and gyrus dentatus neurons were significantly increased in the exercised rats. There was no significant difference of CA2 neuron density between exercise and control groups. There was no significantly differences in any groups according to the results of apoptosis that account of TUNEL positive cells. The present results suggest that regular moderate aerobic treadmill exercise benefit in cognitive functions. This result may derive from treadmill exercise-induced increase cell density without altering of apoptosis in the hippocampus and dentate gyrus of adolescent rats. PMID- 15878800 TI - Comment on: Central neuropathic itch from spinal-cord cavernous hemangioma: a human case, a possible animal model, and hypotheses about pathogenesis, Dey et al., Pain 113 (2005) 233-237. PMID- 15878801 TI - Reversal of the circadian expression of tyrosine-hydroxylase but not nitric oxide synthase levels in the spinal cord of dopamine D3 receptor knockout mice. AB - Circadian rhythms have been described for numerous transmitter synthesizing enzymes in the brain but rarely in spinal cord. We measured spinal tyrosine hydroxylase (TH) and nitric oxide synthase (NOS) levels in the thoracic intermediolateral nucleus, the location of sympathetic preganglionic neurons, in male wild type (WT) and dopamine D(3) receptor knockout mice (D(3)KO). TH and NOS levels both displayed circadian patterns in WT and D(3)KO animals with overall reduced TH and increased NOS expression in the D(3)KO mice. The circadian pattern of NOS expression was similar in WT and D(3)KO mice. In contrast, TH expression was inverted in D(3)KO mice, with TH levels consistently lower than in WT throughout the day, but strongly increased temporarily 1 h prior to daylight. TH is the rate-limiting enzyme for the production of dopamine. Spinal dopamine dysfunction is implicated in a sleep disorder called restless legs syndrome (RLS). RLS follows a circadian rhythm and is relieved clinically by dopamine D(3) receptor agonists. Our observations of an altered circadian pattern in spinal dopamine synthesis in D(3)KO animals may provide insight into putative dopaminergic mechanisms contributing to RLS. PMID- 15878802 TI - The lateral amygdala processes the value of conditioned and unconditioned aversive stimuli. AB - The amygdala is critical for acquiring and expressing conditioned fear responses elicited by sensory stimuli that predict future punishment, but there is conflicting evidence about whether the amygdala is necessary for perceiving the aversive qualities of painful or noxious stimuli that inflict primary punishment. To investigate this question, rats were fear conditioned by pairing a sequence of auditory pips (the conditioned stimulus, or CS) with a brief train of shocks to one eyelid (the unconditioned stimulus, or US). Conditioned responding to the CS was assessed by measuring freezing responses during a test session conducted 24 h after training, and unconditioned responding to the US was assessed by measuring head movements evoked by the eyelid shocks during training. We found that pre training electrolytic lesions of the amygdala's lateral (LA) nucleus blocked acquisition of conditioned freezing to the CS, and also significantly attenuated unconditioned head movements evoked by the US. Similarly, bilateral inactivation of the amygdala with the GABA-A agonist muscimol impaired acquisition of CS evoked freezing, and also attenuated US-evoked responses during training. However, when amygdala synaptic plasticity was blocked by infusion of the NR2B receptor antagonist ifenprodil, acquisition of conditioned freezing was impaired but shock reactivity was unaffected. These findings indicate that neural activity within the amygdala is important for both predicting and perceiving the aversive qualities of noxious stimuli, and that synaptic plasticity within LA is the mechanism by which the CS becomes associated with the US during fear conditioning. PMID- 15878803 TI - Time-dependent changes in extracellular glutamate in the rat dorsolateral striatum following a single cocaine injection. AB - Acute cocaine administration has been shown to alter dorsal striatal plasticity [Proc Natl Acad Sci USA 87 (1990) 6912; Brain Res Bull 30 (1993) 173] and produce long-term neurochemical changes [Pharmacol Biochem Behav 27 (1987) 533]. To date, the effects of acute cocaine on extracellular glutamate and nerve terminal glutamate immunolabeling in the rat dorsolateral striatum have not been reported. To investigate cocaine-induced changes in extracellular glutamate, in vivo microdialysis was carried out in the dorsolateral striatum of rats 1-14 days after receiving a single injection of either vehicle or 15 mg/kg cocaine. There was an increase in the group injected with cocaine 1 day prior to measuring extracellular glutamate as compared with the control group. The group injected with cocaine 3 days prior to the microdialysis session had decreased extracellular glutamate levels. Furthermore, extracellular glutamate remained attenuated 14 days after acute cocaine treatment. Striatal glutamate decreased in the cocaine-treated rats after calcium removal, suggesting that cocaine-induced changes in extracellular glutamate were partially calcium-dependent. The density of nerve terminal glutamate immunolabeling was measured using immunogold electron microscopy in the contralateral striatum of the same rats that had been acutely treated with cocaine or vehicle. There were no changes in the density of glutamate immunolabeling within identified nerve terminals making an asymmetrical (excitatory) synaptic contact 1, 2, 3, or 14 days after acute cocaine exposure as compared with the control groups. Hence, these alterations in extracellular glutamate did not result from changes in glutamate immunolabeling within the synaptic vesicle pool. In addition, no changes in glutamate immunolabeling were found in rats that received cocaine 2 h previously or were withdrawn after 1 week of cocaine administration. The results demonstrate that a single injection of cocaine produces biphasic, time-dependent changes in extracellular glutamate in the rat dorsolateral striatum. PMID- 15878804 TI - Hippocalcin-deficient mice display a defect in cAMP response element-binding protein activation associated with impaired spatial and associative memory. AB - Hippocalcin is a member of the neuronal calcium sensor (NCS) protein family that is highly expressed in hippocampal pyramidal cells and moderately expressed in the neurons of cerebral cortex, cerebellum and striatum. Here we examined the physiological roles of hippocalcin using targeted gene disruption. Hippocalcin deficient (-/-) mice displayed no obvious structural abnormalities in the brain including hippocampal formation at the light microscopic level. Deletion of hippocalcin did not result in up-regulation of the hippocalcin-related proteins; neural visinin-like Ca(2+)-binding proteins (NVP) 1, 2, and 3. The synaptic excitability of hippocampal CA1 neurons appeared to be normal, as estimated by the shape of field excitatory postsynaptic potentials elicited by single- and paired-pulse stimuli, and by tetanic stimulation. However, N-methyl-d-aspartate stimulation- and depolarization-induced phosphorylation of cAMP-response element binding protein (CREB) was significantly attenuated in -/- hippocampal neurons, suggesting an impairment in an activity-dependent gene expression cascade. In the Morris water maze test, the performance of -/- mice was comparable to that of wild-type littermates except in the probe test, where -/- mice crossed the previous location of the platform significantly less often than +/+ mice. Hippocalcin-deficient mice were also impaired on a discrimination learning task in which they needed to respond to a lamp illuminated on the left or right side to obtain food reinforcement. No abnormalities were observed in motor activity, anxiety behavior, or fear learning. These results suggest that hippocalcin plays a crucial role in the Ca(2+)-signaling pathway that underlies long-lasting neural plasticity and that leads to spatial and associative memory. PMID- 15878805 TI - Signal transduction pathways of nitric oxide release in primary microglial culture challenged with gram-positive bacterial constituent, lipoteichoic acid. AB - Between one-third and one-half of all cases of sepsis are known to be caused by gram-positive microorganisms through the cell wall component, e.g. lipoteichoic acid (LTA). Gram-positive bacteria are also known to induce encephalomyelitis and meningeal inflammation, and enhance the production of nitric oxide (NO) via expression of inducible nitric oxide synthase (iNOS) in murine tissue macrophages. It remains to be explored if LTA could activate microglia considered to be resident brain macrophages. We report here that LTA derived from gram positive bacteria (Staphylococcus aureus) significantly induces NO release and iNOS expression in primary microglia. LTA-induced NO accumulation was detected at 2 h in microglial culture and was significantly attenuated by pretreatment with anti-CD14, complement receptor type 3 (CR3) or scavenger receptor (SR) antibodies. LTA activated mitogen-activated protein kinases (MAPKs) such as extracellular signal-regulated kinase, p38 MAPK or c-Jun N-terminal kinase in cultured microglia. LTA-elicited microglial NO production was also drastically suppressed by SB203580 (p38 MAPK inhibitor) or pyrrolidine dithiocarbamate (an inhibitor of nuclear factor kappaB), indicating that p38 MAPK and nuclear factor kappaB were involved in microglial NO release after LTA challenge. These results suggest that gram-positive bacterial product such as LTA can activate microglia to release NO via the signal transduction pathway involving multiple LTA receptors (e.g. CD14, CR3 or SR), p38 MAPK and nuclear factor kappaB. The in vivo study further confirmed that administered intracerebrally LTA induced considerable noticeable iNOS, phospho-IkappaB and phospho-p38 MAPK expression in microglia/macrophages. PMID- 15878806 TI - Neonatal lesion of forebrain cholinergic neurons: further characterization of behavioral effects and permanency. AB - Intraventricular injections of 192 IgG-saporin in the neonatal rat caused severe loss of basal forebrain cholinergic neurons and ectopic hippocampal ingrowths. These were evident at 24 months of age and thus, were lifelong consequences of the 192 IgG-saporin treatment. When tested as young adults on a novel water escape radial arm maze, the rats with this lesion were slower to learn the task, committing significantly more working and reference memory errors before they achieved control level of performance. It is unlikely that this was a result of attentional impairment as the lesioned rats performed as vigilantly as controls in a five choice serial reaction time task. When tested in the Morris water maze at 22 months of age, they were slower at learning the hidden platform location. This contrasts with previous studies which have repeatedly shown that they normally acquire this task as young adults. It was concluded that this neonatal cholinergic lesion has modest but discernable effects on problem solving in young adulthood that are consistent with the reported effects of the lesion on cortical pyramidal neurons. The cognitive effects of the lesion may become more severe in aging, perhaps as a result of the added effects of aging on these neurons. PMID- 15878807 TI - Activating transcription factor 2 expression in the adult human brain: association with both neurodegeneration and neurogenesis. AB - Activating transcription factor 2 (ATF2) is a member of the activator protein-1 family of transcription factors, which includes c-Jun and c-Fos. ATF2 is highly expressed in the mammalian brain although little is known about its function in nerve cells. Knockout mouse studies show that this transcription factor plays a role in neuronal migration during development but over-expression of ATF2 in neuronal-like cell culture promotes nerve cell death. Using immunohistochemical techniques we demonstrate ATF2 expression in the normal human brain is neuronal, is found throughout the cerebral cortex and is particularly high in the granule cells of the hippocampus, in the brain stem, in the pigmented cells of the substantia nigra and locus coeruleus, and in the granule and molecular cell layers of the cerebellum. In contrast to normal cases, ATF2 expression is down regulated in the hippocampus, substantia nigra pars compacta and caudate nucleus of the neurological diseases Alzheimer's, Parkinson's and Huntington's, respectively. Paradoxically, an increase in ATF2 expression was found in the subependymal layer of Huntington's disease cases, compared with normal brains; a region reported to contain increased numbers of proliferating progenitor cells in Huntington's disease. We propose ATF2 plays a role in neuronal viability in the normal brain, which is compromised in susceptible regions of neurological diseases leading to its down-regulation. In contrast, the increased expression of ATF2 in the subependymal layer of Huntington's disease suggests a role for ATF2 in some aspect of neurogenesis in the diseased brain. PMID- 15878808 TI - Outpatient alcoholism treatment: predictors of outcome after 3 years. AB - AIMS: This prospective study investigated predictors for relapse 3 years after completion of an intensive outpatient treatment programme for alcoholism. DESIGN: As previous studies mainly revealed that severity of alcohol dependence, and comorbid psychopathology were predictive for subsequent relapses, the impact of these and other pre-treatment variables on the 36-month outcome was evaluated in a logistic regression analysis. A structured interview was used to assess the variables. Patients were personally interviewed at entry to, and the end of, an outpatient treatment programme, and 6, 12, 24 and 36 months after the end of treatment. One hundred and three alcohol-dependent participants who were taking part in an outpatient treatment were consecutively recruited. RESULTS: Seventy four patients completed the treatment programme. At the follow-up after 36 months, 2 patients had died (after heavy alcohol relapse) and 88 (88%) of the remaining patients could be located and personally re-interviewed. Forty-four (43%) patients were abstinent, 46 (45%) had relapsed and 12 (12%) were classified as improved for the total follow-up period according to the classification proposed by Feuerlein and Kufner. Based on a logistic regression analysis, significant variables for prediction of relapse were treatment drop-outs, female sex and sum of positive life events prior to treatment (relapsers had significantly fewer positive life events). CONCLUSIONS: In contrast to previous studies we could not confirm the importance of determinants known as risk factors for relapse like severity of alcohol dependence. The strongest predictor for relapsing after treatment is treatment drop-out. Since women were at an increased risk for relapse gender-specific treatment approaches should be considered. In summary, the effectiveness of the studied intensive outpatient treatment programme, with an abstinence rate of 43% for the total follow-up period of 3 years, is favourable although selection criteria of must be taken into account. PMID- 15878809 TI - Organisation of trypsin genes in the salmon louse (Lepeophtheirus salmonis, Crustacea, copepoda) genome. AB - Trypsins constitute a subclass of the S1A family of serine peptidases found in all groups of animal and some bacteria. At present, no information about the genomic organisation of trypsins is available for copepods. The only data of copepod trypsins indicate several different trypsins in the marine parasitic copepod Lepeophtheirus salmonis. In the present study, 31.7 kbp of genomic DNA surrounding the previously described LsTryp1-5 sequences was sequenced. The sequenced regions contain nine full-length and three partial trypsin genes. A conservative estimate based on PCR analysis and genomic sequence indicated at least 22 different trypsin genes in L. salmonis, of which 18 are most similar to the previously described LsTryp1 and -2 cDNA sequences. Four of these genes are putative pseudogenes. In addition, a putative mariner like transposase gene was identified. The genomic sequences suggest that the L. salmonis trypsin genes reside within one or more gene clusters. Three different LsTryp intron exon structures were identified, and all three are different from the intron exon organisation previously reported for other S1A peptidases. This implies several intron loss and gain events in the evolution of the L. salmonis trypsin genes. PMID- 15878810 TI - Calorimetric study of bovine serum albumin dilution and adsorption onto polystyrene particles. AB - Titration calorimetry was used to investigate the interaction between a model antigen, bovine serum albumin (BSA), and a model particulate carrier, polystyrene (PS). The binding enthalpy was much higher than reported in the literature for a similar system and did not display a sigmoidal binding curve. These experiments may have accessed low coverage surface sites due to the irreversible nature of protein binding and stepwise titration. An important correction is the heat of dilution of the protein solution. Two regimes were observed: at low concentrations of BSA (below ca. 0.3% (w/v)) an exothermic dilution enthalpy of ca. -100 mJ mg-1 was determined, whereas at higher concentrations of BSA values of ca. -20 mJ mg-1 were obtained. Solution rheological data also showed a change at 0.3% (w/v) BSA, so we hypothesise that the fraction of the BSA as monomers, dimers and polymers in solution changes at approximately 0.3% (w/v). PMID- 15878811 TI - Skin permeation enhancement by sucrose esters: a pH-dependent phenomenon. AB - The purpose of the present study was to evaluate the effect of sucrose esters (particularly, sucrose laureate and sucrose oleate in Transcutol) on the percutaneous penetration of a charged molecule as a function of ionization. We have investigated the influence of these sucrose esters on the in vitro diffusion profiles of lidocaine hydrochloride, a weak ionizable base (pKa=7.9), at different pH values, using porcine ear skin as the barrier membrane. As expected, lidocaine flux in the absence of an enhancer, increased from pH 5 to 9 with a corrresponding increase in the level of the unionized base. However, when skin was pretreated with 2% laureate in Transcutol (2% L-TC), drug permeation was higher at pH 5.0 and 7.0 than at 9.0. A different trend was observed in experiments with 2% oleate in Transcutol (2% O-TC), where skin flux was maximal at a more basic pH, when the degree of ionization is low. The results suggest that sucrose laureate enhances the penetration of the ionized form of the drug (12-fold greater flux relative to control), whereas sucrose oleate is more effective in promoting permeation of the unionized species. The structural properties of the sucrose esters as well as the degree of ionization of the drug are important characteristics affecting the transdermal flux of lidocaine. PMID- 15878812 TI - San-Huang-Xie-Xin-Tang attenuates inflammatory responses in lipopolysaccharide exposed rat lungs. AB - In this study, the potential anti-inflammatory effect of San-Huang-Xie-Xin-Tang (SHXT) and its main component baicalin on LPS-induced lung injury were investigated and compared to the profile of dexamethasone (DEXA) in a pre clinical animal model. Post-treatment with SHXT (75 mg/kg), baicalin (1.5 mg/kg) and DEXA (0.5 mg/kg), significantly inhibited LPS-induced hypotension, lung edema and acute survival rates. Western blotting analysis results indicated that all of them significantly inhibited LPS-induced iNOS, TGF-beta, p38MAPK, and ICAM-1 expressions in the lung tissues. Results from ELISA analysis showed that SHXT, baicalin and DEXA all decreased plasma levels of IL-1beta, TNF-alpha, and MCP-1 caused by LPS. Based on these findings, SHXT and baicalin decreased plasma concentrations of IL-1beta, TNF-alpha, MCP-1, and expressions of TGF-beta, ICAM 1, phosphorylated p38 MAPK, and iNOS, which were associated with lung injury and lethality. These evidences indicated that SHXT and baicalin showed strong anti inflammatory activity, similar to that observed for DEXA, and therefore implicated that herbal SHXT might be therapeutically useful for the treatment of endotoxic lung injury. PMID- 15878813 TI - Fatal steamer accident; blunt force injuries and drowning in post-mortem MSCT and MRI. AB - Steamer accidents, through contact with the bucket wheel, are very seldom today. No publication of such a kind of fatal accident could be found in literature. We present the case of a fatal steamer accident, in which the findings of a blunt traumatization of a person by the ship was completely documented by post-mortem combined multi-slice computed tomography (MSCT) and magnetic resonance imaging (MRI) examinations. A rupture of the aorta was detected using both radiological methods without use of radiopaque material. Radiological examination revealed a comminuted fracture of the thorax vertebrae at the same level as the aortic rupture. Injuries of the soft tissues of the back, caused by the bucket wheel of the steamer, were also diagnosed. In addition to the signs of blunt force trauma the findings of drowning such as an over inflation of the lungs, fluid in the stomach and duodenum were revealed. Furthermore, algological analysis detected diatoms in the lung tissue and blood from the left heart. Therefore, the cause of death was considered being a combination of fatal hemorrhage, caused by the aortic rupture, and drowning. We conclude that virtual autopsy using combined post-mortem MSCT and MRI is a useful tool for documentation, visualisation and analysis of the findings of blunt force trauma and drowning with a large potential in forensic medicine. PMID- 15878814 TI - Inferring the population of origin of DNA evidence within the UK by allele specific hybridization of Y-SNPs. AB - Marked differences in Y-SNP allele frequencies between continental populations can be used to predict the biogeographic origin of a man's ancestral paternal lineage. Using 627 samples collected from individuals within the UK with pale skinned Caucasian, dark-skinned Caucasian, African/Caribbean, South Asian, East Asian or Middle Eastern appearance we demonstrate that an individual's Y-SNP haplogroup is also strongly correlated with their physical appearance. Furthermore, experimental evaluation of the Marligen Signet Y-SNP kit in conjunction with the Luminex 100 detection instrument indicates that reliable and reproducible haplogrouping results can be obtained from 1 ng or more of target template derived from a variety of forensic evidence types including, blood, saliva and post-coital vaginal swabs. The test proved highly male-specific with reliable results being generated in the presence of a 1000-fold excess of female DNA, and no anomalous results were observed during degradation studies despite a gradual loss of typable loci. Hence, Y-SNP haplogrouping has considerable potential forensic utility in predicting likely ethnic appearance. PMID- 15878815 TI - [Tularemia. The disease and its epidemiology in France]. AB - OBJECTIVE: This study had for aim to determine whether the epidemiologic features of animal and human tularemia observed in France could explain its re-emergence since 1993. MATERIALS AND METHODS: A retrospective study was made of outbreaks in animals and human cases and contamination. RESULTS: Twenty to sixty animal outbreaks were recorded every year but the number of cases was under-estimated. This indicates an increase in animal cases. Human cases or contaminations were associated with areas where animal cases had been recorded, except for a few cases more difficult to explain. Several factors are implicated: the disease is not known, except by hunters. Tularemia is more often described in hares but the disease may be present in other species. The disease and its forms should be documented for clinicians and biologists for an adequate clinical and bacteriological diagnosis. COMMENTS: Several factors in the epidemiology of the animal diseases have changed since 1993. It is necessary to be increasingly vigilant, for animal cases and human contamination. It is necessary to strengthen medical and veterinary supervision especially since Francisella tularensis may be used for bioterrorism. PMID- 15878816 TI - [General practice and surveillance: the example of influenza in France]. AB - Data registration by the GROG national network (Regional Group for the Surveillance of Influenza) since 1984 has helped to identify methodological problems. The choice of sentinels and the selection of indicators depend on the analysis of actual influenza consequences. Various sentinels may be concerned: health insurance companies, private companies, schools, physicians, pharmacists. Health care organization modifies the validity of indicators. In France, for instance, home visits were an excellent indicator for early warning before 1995 but this indicator is no longer as efficient as before. The virological detection of Influenza depends on the organization of transportation (samples, results). The predictive value of cases definitions depends on the incidence of infection. The level of choice between specificity and sensibility modifies the perception of outbreaks. Sentinel participation rate influences the sample representativity. The farther this rate is from 100%, the more result validity decreases (in the same way that patients lost to follow-up compromise the validity of results in clinical trials). The publication of results can modify health expenses and behaviors. CONCLUSION: The GROG network stresses the important role that general practitioners play in health surveillance; it also raises questions in the field of mathematics, statistics, professional organization, training, education, and politics. PMID- 15878817 TI - [Sexually transmitted infections. Current clinical and therapeutic data]. AB - A recent increase of syphilis and gonorrhea has been observed in France. More recently, the resurgence of very infrequent STDs, such as lymphogranuloma venerum, has been noticed in Western Europe. This data illustrates the emergence of high-risk behavior and the relative failure of prevention for sexually transmitted infection and HIV infection, especially in homosexual men. Since the onset of the AIDS epidemic, oral sex has been increasingly used both by heterosexual and homosexual partners. Even if the risk of HIV transmission with oral sex is very low, oral sex is a major mode of transmission for syphilis, gonorrhea, and genital herpes. Condoms are seldom used for oral sex. The transmission of STI by oral sex is less documented in heterosexual than in homosexual patients. The level of knowledge does not seem to reduce risky behavior in the most concerned population. An infection not normally considered as sexually transmitted (i.e. hepatitis C) may be transmitted by high risk sexual behavior increasing the risk of blood-blood contact, as recently reported among HIV-infected homosexual men. The emergence of high-risk sexual practices (traumatic practices, fisting) in a specific population required focused prevention measures to avoid blood-transmitted infections. The increased risk of HIV transmission by ST co-infection (syphilis, genital herpes, gonorrhea) and the potential morbidity of STI in HIV infected patients should also be emphasized. PMID- 15878818 TI - [Acute transverse myelitis associated with Mycoplasma pneumoniae infection in an eight year old boy]. AB - Many Mycoplasma pneumoniae extra respiratory infections have already been reported. The authors report the case of an eight year old child, presenting with acute transverse myelitis, with a PCR proven involvement of M. pneumoniae in CSF as well as the presence of specific IgM in blood. Acute transverse myelitis may have many causes, most of the time viral. The mechanisms of neurological involvement in M. pneumoniae infections are still unclear, but several points indicate an immune reaction. Corticotherapy first i.v. then per os has proved to be an efficient treatment for acute myelitis. Antibiotherapy is discussed because of the undocumented mechanisms of neurological involvement related to the mycoplasma. PMID- 15878819 TI - [Antiplatelet agents and regional anaesthesia: experience in 130 patients]. PMID- 15878820 TI - Pathological mandibular fractures following third molar removal. AB - Seventeen patients, who presented mandibular fractures following third molar removal, were treated in the Department of Oral and Maxillofacial Surgery at the University Hospital Freiburg between 2000 and 2004. Clinical and radiological data were analysed to evaluate the possible risk of fracture following third molar removal. Our data show an increased rate of pathological mandibular fractures in males over 40 years of age, following removal of a difficult lower wisdom tooth with a high retention grade that necessitated bone removal. Fourteen out of 17 fractures occurred postoperatively. In six cases, no fracture was visible in radiographs during the primary investigation. Pathological mandibular fractures were typically located anterior to the mandibular angle (n=15). We recommend informing patients about possible risks and, in selective cases, a soft diet for up to 4 weeks after the operation. A cracking noise reported by the patient is the most important indication of a fracture. If, initially, the fracture is radiologically undetectable, we recommend the use of radiological controls. PMID- 15878821 TI - Changes in condylar long axis and skeletal stability after bilateral sagittal split ramus osteotomy with poly-L-lactic acid or titanium plate fixation. AB - This study was designed to assess skeletal stability after bilateral sagittal split ramus osteotomy (BSSO) and fixation with a poly-l-lactic acid (PLLA) plate, as compared to that after BSSO and fixation with a titanium plate, and to analyze the change in the condylar long axis after these procedures. The study group comprised 40 patients who had mandibular prognathism (20, titanium group; 20, PLLA group). The groups were randomized to show similar distributions of preoperative SNB. All patients underwent BSSO setback by the Obwegeser method. Fixation was done with bent titanium plates or bent PLLA plates, applied in a similar manner. Lateral, frontal, and submental-vertical cephalograms were analyzed preoperatively and postoperatively. The maximum mouth opening range and the incidence of temporomandibular disorders were also evaluated. There was no significant difference in the right condylar angle or width between the two groups, but the left condylar angle and width, gonial angle, and ramus inclination differed significantly between them (P<0.05). SNA, SNB, and ANB were similar in both groups. There was no significant difference between the groups in maximum mouth opening range or temporomandibular disorders. We conclude that the change in condylar angle after BSSO and fixation with a titanium plate is greater than that after BSSO and fixation with a PLLA plate, but skeletal stability related to the occlusion is similar for the two procedures. PMID- 15878822 TI - Rise in ciprofloxacin resistance in Escherichia coli from urinary tract infections from 1999-2004. PMID- 15878823 TI - [X-linked adrenoleukodystrophy in a female proband: clinical presentation, biological diagnosis and family consequences]. AB - INTRODUCTION: X-linked adrenoleukodystrophy (ALD) is the most frequent type of leukodystrophy (1/17 000 males). The phenotypic range in male patients varies from the severe cerebral presentations in children to the milder myeloneuropathy and to isolate adrenal insufficiency. More than a half of the carrier females display clinical symptoms over the age of 40 years. OBSERVATION: Diagnosis of ALD was raised in a 40 year-old female who presented with spastic paraparesis and sphincterian dysfunction, occurring after the delivery of her first child. There was no family history of ALD. Very long-chain fatty acids (VLFCA) were assayed in her one-year-old son in order to propose appropriate hormonal and neurological survey. His dosage was abnormal and an adrenal insufficiency was subsequently found. A brain MRI will be proposed biannually when he reaches to age of for years. The proband's mother had an increased level of VLCFA, showing that she was a carrier. Family screening was extended to the proband's sisters and maternal aunt who already had children, but also to her brother, who may express a mild form of the disease later on, and to her maternal uncles who might be asymptomatic carriers. A frameshift mutation was found in the ABCD1 gene and will allow accurate carrier identification and prenatal diagnosis in the family. CONCLUSION: ALD diagnosis should be evoked in a woman affected by myelopathy despite the lack of family history. Such a diagnosis has severe consequences since some of the related males may carry the mutation although they do not display any symptom at time of diagnosis, and because carrier females have a risk to both have a clinical expression of the disease and give birth to an affected boy. PMID- 15878824 TI - [The biochemical diagnosis of Gaucher disease]. AB - Gaucher disease is a disease of overload lyosomale which we often met since a score of years. Since 1980 we had to answer several requests for diagnosis of this metabolic disease. Requests emanating primarily from paediatric services. Twelve cases were confirmed within sight of measurement of the intra-leucocytic activity of the beta-glucocerebrosidase, enzyme intervening in the catabolism of the sphingolipides. We report here our experiment in the biochemical diagnosis of Gaucher disease by showing mainly the variability and the extreme heterogeneity of the activity of the beta-glucosidasic during practised measurements. In addition, we expose the problems of diagnosis etiologic which certain patients raise in front of the discordances between the measured enzymatic activity and clinical signs of the disease of left-handed person. In addition, we develop the biological parameters useful to proportion for the monitoring of the treatment which is finally available in our country. PMID- 15878825 TI - A mixed self-assembled monolayer-based surface plasmon immunosensor for detection of E. coli O157:H7. AB - The sensitivity and specificity of a polyethylene glycol terminated alkanethiol mixed self-assembled monolayers (SAM) on surface plasmon resonance (SPR) immunosensor to detect Escherichia coli O157:H7 is demonstrated. Purified monoclonal (Mabs) or polyclonal antibodies (PAbs) against E. coli O157:H7 were immobilized on an activated sensor chip and direct and sandwich assays were carried to detect E. coli O157:H7. Effect of Protein G based detection and effect of concentrations of primary and secondary antibodies in sandwich assay were investigated. The sensor surface was observed under an optical microscope at various stages of the detection process. The sensor could detect as low as 10(3)CFU/ml of E. coli O157:H7 in a sandwich assay, with high specificity against Salmonella Enteritidis. The detection limit using direct assay and Protein G were 10(6)CFU/ml and 10(4)CFU/ml, respectively. Results indicate that an alkanethiol SAM based SPR biosensor has the potential for rapid and specific detection of E. coli O157:H7, using a sandwich assay. PMID- 15878826 TI - Analogs of 1alpha,25-dihydroxyvitamin D(3) as novel inhibitors of renin biosynthesis. AB - The renin-angiotensin system (RAS) plays a central role in the pathogenesis of hypertension. Recently, we discovered that 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] functions as a negative endocrine regulator of renin biosynthesis, which provides a molecular basis to explore the potential of Vitamin D analogs as renin inhibitors to control the RAS. To search for renin inhibiting Vitamin D analogs, we screened 20 Vitamin D analog compounds using As4.1-hVDR cell (a juxtaglomerular cell line) culture by Northern blot and luciferase reporter assays. We found that the Gemini compounds, which have two side-chains at carbon-20 position, were particularly active in suppressing renin expression. Eight Gemini compounds were identified that were equally or 10- to 100-times more potent than 1,25(OH)(2)D(3) in renin inhibition. These Gemini compounds also potently stimulate 25-hydroxyvitamin D 24-hydroxylase expression in As4.1-hVDR cells. Administration of compound RO-27-5646 [1,25-dihydroxy-21-(3 methyl-3-hydroxy-butyl)-19-nor-cholecalciferol] in mice caused a marked reduction in renal renin mRNA expression without invoking severe hypercalcemia as seen in 1,25(OH)(2)D(3) treatment. These data establish in principle that Vitamin D analogs can indeed inhibit renin expression in vitro and in vivo, and support the notion that low calcemic Vitamin D analogs can potentially be used as therapeutic agents to control the RAS. PMID- 15878827 TI - Expression, purification, and functional testing of recombinant CYR61/CCN1. AB - The human cysteine-rich protein 61 (CYR61/CCN1) belongs to the CCN family of genes which plays an important role in cellular processes such as proliferation, migration, adhesion, and differentiation. These extracellular matrix signaling molecules consist of a modular structure and contain 38 conserved cysteine residues. Previously, we have shown that CYR61 is expressed in human osteoblasts and is regulated by bone-relevant growth factors. The protein also plays a role in angiogenesis. The open reading frame was cloned into a baculovirus expression vector and transfected into SF-21 insect cells. Recombinant protein was expressed as a fusion protein with the Fc-domain of human IgG and purified using affinity chromatography on protein G-Sepharose columns. The chorioallantoic membrane assay verified that blood vessel formation was stimulated by rCYR61. Additionally, human primary mesenchymal stem cells, osteoblasts, and endothelial cells responded to CYR61 treatment by a markedly stimulated proliferation. rCYR61-Fc represents a tool to elucidate its role in cells of the bone microenvironment. PMID- 15878828 TI - A rapid and efficient protocol to purify biologically active recombinant proteins from mammalian cells. AB - Here, we describe a simple and efficient method for the expression and purification of active recombinant proteins in mammalian cells. This method uses the expression of T7 epitope-tagged proteins in transiently transfected 293T cells grown in monolayer, followed by anti-T7-agarose affinity chromatography. This procedure yields approximately between 75 and 100 microg of biologically active protein/150 cm(2) flask that can be used for biochemical studies. We have tested this protocol for the expression of the prototype SR protein, SF2/ASF, which is a member of the SR protein family with a role in constitutive and alternative splicing. We show that SF2/ASF purified using this protocol is able to complement an S100 HeLa extract, demonstrating that is biologically active. Moreover, expression of a novel SR-related protein that it is required for the second step of pre-mRNA splicing also rendered an active protein. In summary, we present a protocol based on transient transfection of mammalian cells that results in easy purification of significant amounts of biologically active proteins. PMID- 15878829 TI - The phylogenetic status of Paxillosida (Asteroidea) based on complete mitochondrial DNA sequences. AB - One of the most important issues in asteroid phylogeny is the phylogenetic status of Paxillosida. This group lacks an anus and suckers on the tube feet in adults and does not develop the brachiolaria stage in early development. Two controversial hypotheses have been proposed for the phylogenetic status of Paxillosida, i.e., Paxillosida is primitive or rather specialized in asteroids. In this study, we determined the complete mitochondrial DNA nucleotide sequences from two paxillosidans (Astropecten polyacanthus and Luidia quinaria) and one forcipulatidan (Asterias amurensis). The mitochondrial genomes of the three asteroids were identical with respect to gene order and transcription direction, and were identical to the previously reported mitochondrial genomes of Asterina pectinifera (Valvatida) and Pisaster ochraceus (Forcipulatida) in this respect. Therefore, the comparison of genome structures was uninformative for the purposes of asteroid phylogeny. However, molecular phylogenetic analyses based on the amino acid sequences and the nucleotide sequences from the five asteroids supported the monophyly of the clade that included the two paxillosidans and Asterina. This suggests that the paxillosidan characters are secondarily derived ones. PMID- 15878830 TI - Optimization and validation of small quantity RNA profiling for identifying TNF responses in cultured human vascular endothelial cells. AB - INTRODUCTION: Affymetrix oligonucleotide microarrays are widely used in basic and applied research (Lander, E.S., (1999). Array of hope. Nature Genetics 21, 3-4; Lockhart, D.J. & Winzeler, E.A. (2000) Genomics, gene expression and DNA arrays. Nature 405, 827-836.) The need for a significant amount of starting RNA has limited its use in applications where the amount of RNA is limiting, such as with Laser Captured Microdissection (LCM), small biopsies, or peripheral blood in rodent models. To overcome this limitation, various RNA amplification and labeling methods have been described, however, further optimization and validation of these methods are needed. METHODS: Here we reported using the Arcturus technology to optimize amplification and labeling of small amounts of RNA for Affymetrix microarray studies. We assessed the technical feasibility and variation introduced by differences in starting RNA quantity and differences in technical performance by microarray hybridization. RESULTS: We demonstrated that the current approach is reliable to amplify as little as 40 ng total RNA, and it is suitable for Affymetrix studies yielding satisfactory quantitative chip performance. We also showed that differences in labeling methods contribute more to variation than the differences in starting RNA quantity per se. As a model, we studied the well-documented TNF-induced inflammatory responses in cultured human vascular endothelial cells. We were able to recapitulate the TNF-induced responses using small RNA sample profiling. PMID- 15878831 TI - Aflatoxin conducive and non-conducive growth conditions reveal new gene associations with aflatoxin production. AB - Research on aflatoxin (AF) production has traditionally focused on defining the AF biosynthetic pathway with the goal of identifying potential targets for intervention. To understand the effect of nitrogen source, carbon source, temperature, and pH on the regulation of AF biosynthesis, a targeted cDNA microarray consisting of genes associated with AF production over time was employed. Expression profiles for genes involved in AF biosynthesis grouped into five clades. A putative regulon was identified consisting of 20 genes that were induced in the conducive nitrogen and pH treatments and the non-conducive carbon and temperature treatments, as well as four other putative regulons corresponding to each of the four variables studied. Seventeen genes exhibited consistent induction/repression profiles across all the experiments. One of these genes was consistently downregulated with AF production. Overexpression of this gene resulted in repression of AF biosynthesis. The cellular function of this gene is currently unresolved. PMID- 15878832 TI - Predictive Bayesian neural network models of MHC class II peptide binding. AB - We used Bayesian regularized neural networks to model data on the MHC class II binding affinity of peptides. Training data consisted of sequences and binding data for nonamer (nine amino acid) peptides. Independent test data consisted of sequences and binding data for peptides of length 0.8. We also used both amino acid indicator variables (bin20) and property-based descriptors to generate models for MHC class II-binding of peptides. The property based descriptors were more parsimonious than the indicator variable descriptors, making them applicable to larger peptides, and their design makes them able to generalize to unknown peptides outside of the training space. None of the external test data sets contained any of the nonamer sequences in the training sets. Consequently, the models attempted to predict the activity of truly unknown peptides not encountered in the training sets. Our models were well able to tackle the difficult problem of correctly predicting the MHC class II-binding activities of a majority of the test set peptides. Exceptions to the assumption that nonamer motif activities were invariant to the peptide in which they were embedded, together with the limited coverage of the test data, and the fuzziness of the classification procedure, are likely explanations for some misclassifications. PMID- 15878833 TI - Glucokinase in chicken (Gallus gallus). Partial cDNA cloning, immunodetection and activity determination. AB - Chickens are more hyperglycaemic and insulin-resistant than mammals, and in efforts to understand their glucose metabolism we investigated whether glucokinase (GK) is present in chicken liver or pancreas. This enzyme plays a major role in glucose-sensing in mammals and we have examined whether it also contributes to glucose homeostasis in chickens. Using RT-PCR, we cloned and sequenced a partial cDNA fragment (750 bp) from liver and pancreas that showed a high degree of identity with mammalian GK. Using antibodies directed towards human GK, we immunodetected a 50 kDa band in chicken liver and pancreas. The molecular mass of the band and its specific interaction with the antibody suggest that this protein corresponds to a chicken homologue of human GK. We also determined by spectrophotometry a glucokinase-like activity in crude liver homogenates with an apparent half saturating concentration for glucose of 8.6 mM. GK gene and protein expression did not differ between fed and 24 h fasted states but GK-like activity was significantly increased in fed chickens. In conclusion, our study provides evidence for the presence of GK gene and protein in chicken liver and pancreas and shows that the liver enzyme is active. PMID- 15878834 TI - The use of animal models to guide rational vaccine design. AB - Although there are several varieties of animal models of tuberculosis, the mouse and the guinea pig are by far the most validated and useful. These provide information about vaccine-induced protection, immunogenicity, toxicity, and immunopathological effects. There is still much to be learned, however, in terms of rational vaccine design, especially in the context of therapeutic or anti latent vaccine formulations and animal models of these situations. PMID- 15878835 TI - The role of "go no-go" decisions in TB vaccine development. AB - "Go no-go" decisions play a critical role in the product development plan of any TB vaccine at several points. Go no-go decisions are designed to serve the fundamental maxim in vaccine development that killing a bad vaccine project as early as possible is the hallmark of a successful overall program; in development, unlike in basic research, costs and skilled manpower requirements rise exponentially as the program proceeds. The opportunity costs in resources, manpower and equipment utilized on bad vaccine projects are unavailable to other vaccine programs. The go no-go decision is a fundamental piece of the process that balances risk, time and resources. An example of a go no-go decision development program is presented. PMID- 15878836 TI - TB subunit vaccines--putting the pieces together. AB - The search for a new and improved vaccine against tuberculosis (TB) is currently a very active field of research, which in the last 10 years has benefited tremendously from the completed Mycobacterium tuberculosis genome and the progress in molecular biology and computer science. In this review, we discuss how Genomics, Proteomics and Transcriptomics have accelerated the pace of antigen discovery and vaccine development and have changed this field completely, resulting in the identification of a large number of antigens with potential in TB vaccines. The next phase of this work has now started--putting the most relevant molecules back together as fusion molecules and cocktails. This requires carefully monitoring aspects as immunodominance, recognition in different populations as well as vaccine manufacturing. PMID- 15878837 TI - The pivotal role of cytokines in muscle wasting during cancer. AB - The cachectic syndrome, characterized by a marked weight loss, anorexia, asthenia and anemia, is invariably associated with the presence and growth of the tumour and leads to a malnutrition status due to the induction of anorexia or decreased food intake. In addition, the competition for nutrients between the tumour and the host leads to an accelerated catabolic state, which promotes severe metabolic disturbances in the host, including hypermetabolism, which leads to an increased energetic inefficiency. Although the search for the cachectic factor(s) started a long time ago, and although many scientific and economic efforts have been devoted to its discovery, we are still a long way from knowing the whole truth. Present investigation is devoted to unrevealing the different signaling pathways (particularly transcriptional factors) involved in muscle wasting. The main aim of the present review is to summarize and evaluate the different molecular mechanisms and catabolic mediators involved in cancer cachexia since they may represent targets for future promising clinical investigations. PMID- 15878838 TI - Role of oxidative stress, endoplasmic reticulum stress, and c-Jun N-terminal kinase in pancreatic beta-cell dysfunction and insulin resistance. AB - Type 2 diabetes is the most prevalent and serious metabolic disease affecting people all over the world. Pancreatic beta-cell dysfunction and insulin resistance are the hallmark of type 2 diabetes. Normal beta-cells can compensate for insulin resistance by increasing insulin secretion and/or beta-cell mass, but insufficient compensation leads to the onset of glucose intolerance. Once hyperglycemia becomes apparent, beta-cell function gradually deteriorates and insulin resistance aggravates. Under diabetic conditions, oxidative stress and endoplasmic reticulum stress are induced in various tissues, leading to activation of the c-Jun N-terminal kinase pathway. The activation of c-Jun N terminal kinase suppresses insulin biosynthesis and interferes with insulin action. Indeed, suppression of c-Jun N-terminal kinase in diabetic mice improves insulin resistance and ameliorates glucose tolerance. Thus, the c-Jun N-terminal kinase pathway plays a central role in pathogenesis of type 2 diabetes and could be a potential target for diabetes therapy. PMID- 15878839 TI - Discrimination between two steps in the mitochondrial permeability transition process. AB - It is well known that a lag phase generally elapses between the addition of inducers of the mitochondrial permeability transition and the opening of the pore. To advance our present understanding as regards the significance of this phenomenon, we used experimental approaches which are sensitive to different aspects of the permeability transition process. The pore conformation was sensed by the fluorescence anisotropy changes of hematoporphyrin-labelled mitochondria. Membrane permeabilization was ascertained by following the matrix swelling consequent to external solute equilibration. We show that the anisotropy changes of mitochondria-bound hematoporphyrin precede both membrane depolarization (proton permeation) and matrix swelling (solute permeation), thus sensing a step of the permeability transition process that involves the pore in its closed state. We suggest that the opening of the pore is preceded by a structural remodelling of mitochondrial domains containing hematoporphyrin-near, pore regulating histidines. Such a perturbation is strongly inhibited at acidic matrix pH and completely blocked by cyclosporin A. In sucrose-based media the opening of the pore can be strongly delayed, as compared to salt-based media, a fact which probably reflects perturbation of mitochondrial membranes by sugar. We conclude that the mitochondrial permeability transition could be described as an at least two-step process which is mainly regulated by conformational changes of the pore components. PMID- 15878840 TI - Pterostilbene and 3'-hydroxypterostilbene are effective apoptosis-inducing agents in MDR and BCR-ABL-expressing leukemia cells. AB - Pterostilbene and 3,5-hydroxypterostilbene are the natural 3,5-dimethoxy analogs of trans-resveratrol and piceatannol, two compounds which can induce apoptosis in tumor cells. In previous studies we demonstrated the importance of a 3,5 dimethoxy motif in conferring pro-apoptotic activity to stilbene based compounds so we now wanted to evaluate the ability of pterostilbene and 3,5 hydroxypterostilbene in inducing apoptosis in sensitive and resistant leukemia cells. When tested in sensitive cell lines, HL60 and HUT78, 3' hydroxypterostilbene was 50-97 times more potent than trans-resveratrol in inducing apoptosis, while pterostilbene appeared barely active. However, both compounds, but not trans-resveratrol and piceatannol, were able to induce apoptosis in the two Fas-ligand resistant lymphoma cell lines, HUT78B1 and HUT78B3, and the multi drug-resistant leukemia cell lines HL60-R and K562-ADR (a Bcr-Abl-expressing cell line resistant to imatinib mesylate). Of note, pterostilbene-induced apoptosis was not inhibited by the pancaspase-inhibitor Z VAD-fmk, suggesting that this compound acts through a caspase-independent pathway. On the contrary, 3'-hydroxypterostilbene seemed to trigger apoptosis through the intrinsic apoptotic pathway: indeed, it caused a marked disruption of the mitochondrial membrane potential delta psi and its apoptotic effects were inhibited by Z-VAD-fmk and the caspase-9-inhibitor Z-LEHD-fmk. Moreover, pterostilbene and 3'-hydroxypterostilbene, when used at concentrations that elicit significant apoptotic effects in tumor cell lines, did not show any cytotoxicity in normal hemopoietic stem cells. In conclusion, our data show that pterostilbene and particularly 3'-hydroxypterostilbene are interesting antitumor natural compounds that may be useful in the treatment of resistant hematological malignancies, including imatinib, non-responsive neoplasms. PMID- 15878841 TI - Application of bone morphogenetic proteins in spinal fusion. AB - We are now entering an exciting new era in spinal surgery where the inherent osteoinductive capacity of the body has been harnessed for bone formation for therapeutic purposes. Recombinant bone morphogenetic proteins have been extensively studied in both the pre-clinical and clinical arena for spinal fusion with considerable success. The challenges facing spine surgeons now is the development of site-specific carriers and optimal doses for these growth factors. This review highlights the recent advances in this regard. PMID- 15878842 TI - Symptom severity as a predictor of reported differences of prehospital delay between medical records and structured interviews among patients with AMI. AB - BACKGROUND: The time interval from symptom onset to hospital arrival can guide the decision to start reperfusion for patients with acute myocardial infarction (AMI). AIMS: The aims of the study were 1) to examine the consistency and agreement of prehospital delay time between medical record review and structured patient interview and 2) to identify whether symptom severity is an independent predictor of differences in reported prehospital delay between these two data sources. METHODS: In this cross-sectional study, a convenience sample of 155 patients with AMI in Japan was recruited. The time and date of symptom onset were obtained from medical record review and a structured patient interview. The interviewer asked about severity of symptoms, using a scale on 0 to 10 scale, with 0 being "no symptom" and 10 being "the most severe symptoms imaginable". RESULTS: The prehospital delay time from the medical record was significantly shorter than from the structured interview. Perfect agreement of reported prehospital delay time between two data sources was found in 46% of patients. Patients with symptom severity scores of greater than 8 on a 10-point scale were 2.2 times (95% CI: Odds Ratio 1.1 to 5.1) more likely to report different symptom onset time. CONCLUSIONS: Prehospital delay time documented in the medical record was shorter than that identified from a structured interview. In addition, the discrepancy between the two data sources may be larger in those patients with severe AMI symptoms. PMID- 15878843 TI - Structure of the G60A mutant of Ras: implications for the dominant negative effect. AB - Substituting alanine for glycine at position 60 in v-H-Ras generated a dominant negative mutant that completely abolished the ability of v-H-Ras to transform NIH 3T3 cells and to induce germinal vesicle breakdown in Xenopus oocytes. The crystal structure of the GppNp-bound form of RasG60A unexpectedly shows that the switch regions adopt an open conformation reminiscent of the structure of the nucleotide-free form of Ras in complex with Sos. Critical residues that normally stabilize the guanine nucleotide and the Mg(2+) ion have moved considerably. Sos binds to RasG60A but is unable to catalyze nucleotide exchange. Our data suggest that the dominant negative effect observed for RasG60A.GTP could result from the sequestering of Sos in a non-productive Ras-GTP-guanine nucleotide exchange factor ternary complex. PMID- 15878844 TI - Chimeric mutations in the M2 segment of the 5-hydroxytryptamine-gated chloride channel MOD-1 define a minimal determinant of anion/cation permeability. AB - The ionic selectivity of ligand-gated ion channels (LGICs) determines whether receptor activation produces an excitatory or inhibitory response. The determinants of anion/cation selectivity were investigated for a new member of the LGIC superfamily, MOD-1, a serotonin-gated chloride channel cloned from the nematode Caenorhabditis elegans. In common with other anionic LGICs (glycine receptors and GABA(A) receptors), the selectivity triple mutant in the pore forming M2 segment (proline insertion, Ala --> Glu substitution at the central ring, and Thr --> Val at the hydrophobic ring) converted the selectivity of MOD-1 from anionic to cationic. Unlike other LGICs, however, this mutant in MOD-1 was highly selective for K+ over other cations. Subsets of this selectivity triple mutant were studied to define the minimal change required for conversion from anion-permeable to cation-permeable. The double mutant at the central ring of charge (deltaPro-269/A270E) produced a non-selective cation channel. Charge reversal at the central ring alone (A270E) was sufficient to convert MOD-1 to cation-permeable. These results refine the determinants of ion-charge selectivity in LGICs and demonstrate the critical role of the central ring of charge formed by the M2 segments. PMID- 15878845 TI - The foot structure from the type 1 ryanodine receptor is required for functional coupling to store-operated channels. AB - In the present study we have explored structural determinants of the functional interaction between skeletal muscle ryanodine receptor (RyR1) and transient receptor potential channel 1 (TRPC1) channels expressed in Chinese hamster ovary cells. We have illustrated a functional interaction between TRPC1 channels and RyR1 for the regulation of store-operated calcium entry (SOCE) initiated after releasing calcium from a caffeine-sensitive intracellular calcium pool. RNA interference experiments directed to reduce the amount of TRPC1 protein indicate that RyR1 associates to at least two different types of store-operated channels (SOCs), one dependent and one independent of TRPC1. In contrast, bradykinin induced SOCE is completely dependent on the presence of TRPC1 protein, as we have previously illustrated. Removing the foot structure from RyR1 results in normal caffeine-induced release of calcium from internal stores but abolishes the activation of SOCE, indicating that this structure is require for functional coupling to SOCs. The footless RyR1 protein shows a different cellular localization when compared with wild type RyR1. The later protein shows a higher percentage of colocalization with FM-464, a marker of plasma membrane. The implications of the foot structure for the functional and physical coupling to TRPC and SOCs is discussed. PMID- 15878846 TI - The RNA-binding protein HuR binds to acetylcholinesterase transcripts and regulates their expression in differentiating skeletal muscle cells. AB - During myogenic differentiation, acetylcholinesterase (AChE) transcript levels are known to increase dramatically. Although this increase can be attributed in part to increased transcriptional activity, posttranscriptional mechanisms have also been implicated in the high levels of AChE mRNA in myotubes. In this study, we observed that transfection of a luciferase reporter construct containing the full-length AChE 3'-untranslated region (UTR) resulted in significantly higher (5 fold) luciferase activity in differentiated myotubes versus myoblasts. RNA electrophoretic mobility shift assays (REMSAs) performed with a full-length AChE 3'-UTR probe and the AU-rich element revealed that the intensity of RNA-binding protein complexes increased as myogenic differentiation proceeded. Using several complementary approaches including supershift REMSA, mRNA-binding protein pull down assays, and immunoprecipitation followed by reverse transcription-PCR, we found that the mRNA-stabilizing protein HuR interacts directly with AChE transcripts. Stable overexpression of HuR in C2C12 cells increased the expression of endogenous AChE transcripts as well as that of the luciferase reporter construct containing the AChE 3'-UTR. In vitro stability assays performed with protein extracts from these cells versus controls resulted in a slower rate of AChE mRNA decay. The down-regulation of HuR expression mediated through small interfering RNA further confirmed the role of HuR in the regulation of AChE mRNA levels. Taken together, these studies demonstrate that HuR interacts with the AChE 3'-UTR to regulate posttranscriptionally the expression of AChE mRNA during myogenic differentiation. PMID- 15878847 TI - An essential tryptophan of Escherichia coli DnaA protein functions in oligomerization at the E. coli replication origin. AB - In the initiation of bacterial DNA replication, DnaA protein recruits DnaB helicase to the chromosomal origin, oriC, leading to the assemble of the replication fork machinery at this site. Because a region near the N terminus of DnaA is required for self-oligomerization and the loading of DnaB helicase at oriC, we asked if these functions are separable or interdependent by substituting many conserved amino acids in this region with alanine to identify essential residues. We show that alanine substitutions of leucine 3, phenylalanine 46, and leucine 62 do not affect DnaA function in initiation. In contrast, we find on characterization of a mutant DnaA that tryptophan 6 is essential for DnaA function because its substitution by alanine abrogates self-oligomerization, resulting in the failure to load DnaB at oriC. These results indicate that DnaA bound to oriC forms a specific oligomeric structure, which is required to load DnaB helicase. PMID- 15878848 TI - Membrane cholesterol content accounts for developmental differences in surface B cell receptor compartmentalization and signaling. AB - Recent studies argue for an important role for cholesterol in maintaining plasma membrane heterogeneity and influencing a variety of cellular processes, including signaling, adhesion, and permeability. Here, we document that tolerance-sensitive transitional immature B cells maintain significantly lower membrane unesterified cholesterol levels than mature-stage splenic B cells. In addition, the relatively low level of cholesterol in transitional immature B cells impairs compartmentalization of their B cell receptor (BCR) into cholesterol-enriched domains following BCR aggregation and reduces their ability to sustain certain aspects of BCR signaling as compared with mature B cells. These studies establish an unexpected difference in the lipid composition of peripheral transitional immature and mature B cells and point to a determining role for development associated differences in cholesterol content for the differential responses of these B cells to BCR engagement. PMID- 15878849 TI - Identification of the major urinary metabolite of the highly reactive cyclopentenone isoprostane 15-A(2t)-isoprostane in vivo. AB - The cyclopentenone isoprostanes (A(2)/J(2)-IsoPs) are formed in significant amounts in humans and rodents esterified in tissue phospholipids. Nonetheless, they have not been detected unesterified in the free form, presumably because of their marked reactivity. A(2)/J(2)-IsoPs, similar to other electrophilic lipids such as 15-deoxy-Delta(12,14)-prostaglandin J(2) and 4-hydroxynonenal, contain a highly reactive alpha,beta-unsaturated carbonyl, which allows these compounds to react with thiol-containing biomolecules to produce a range of biological effects. We sought to identify and characterize in rats the major urinary metabolite of 15-A(2t)-IsoP, one of the most abundant A(2)-IsoPs produced in vivo, in order to develop a specific biomarker that can be used to quantify the in vivo production of these molecules. Following intravenous administration of 15 A(2t)-IsoP containing small amounts of [(3)H(4)]15-A(2t)-IsoP, 80% of the radioactivity excreted in the urine remained in aqueous solution after extraction with organic solvents, indicating the formation of a polar conjugate(s). Using high pressure liquid chromatography/mass spectrometry, the major urinary metabolite of 15-A(2t)-IsoP was determined to be the mercapturic acid sulfoxide conjugate in which the carbonyl at C9 was reduced to an alcohol. The structure was confirmed by direct comparison to a synthesized standard and via various chemical derivatizations. In addition, this metabolite was found to be formed in significant quantities in urine from rats exposed to an oxidant stress. The identification of this metabolite combined with the finding that these metabolites are produced in in vivo settings of oxidant stress makes it possible to use this method to quantify, for the first time, the in vivo production of cyclopentenone prostanoids. PMID- 15878850 TI - Mitochondrial deoxynucleotide pools in quiescent fibroblasts: a possible model for mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). AB - Mitochondrial (mt) DNA depletion syndromes can arise from genetic deficiencies for enzymes of dNTP metabolism, operating either inside or outside mitochondria. MNGIE is caused by the deficiency of cytosolic thymidine phosphorylase that degrades thymidine and deoxyuridine. The extracellular fluid of the patients contains 10-20 microM deoxynucleosides leading to changes in dTTP that may disturb mtDNA replication. In earlier work, we suggested that mt dTTP originates from two distinct pathways: (i) the reduction of ribonucleotides in the cytosol (in cycling cells) and (ii) intra-mt salvage of thymidine (in quiescent cells). In MNGIE and most other mtDNA depletion syndromes, quiescent cells are affected. Here, we demonstrate in quiescent fibroblasts (i) the existence of small mt dNTP pools, each usually 3-4% of the corresponding cytosolic pool; (ii) the rapid metabolic equilibrium between mt and cytosolic pools; and (iii) the intra-mt synthesis and rapid turnover of dTTP in the absence of DNA replication. Between 0.1 and 10 microM extracellular thymidine, intracellular thymidine rapidly approaches the extracellular concentration. We mimic the conditions of MNGIE by maintaining quiescent fibroblasts in 10-40 microM thymidine and/or deoxyuridine. Despite a large increase in intracellular thymidine concentration, cytosolic and mt dTTP increase at most 4-fold, maintaining their concentration for 41 days. Other dNTPs are marginally affected. Deoxyuridine does not increase the normal dNTP pools but gives rise to a small dUTP and a large dUMP pool, both turning over rapidly. We discuss these results in relation to MNGIE. PMID- 15878851 TI - Cyclic AMP and AKAP-mediated targeting of protein kinase A regulates lactate dehydrogenase subunit A mRNA stability. AB - Expression of the lactate dehydrogenase A subunit (ldh-A) gene is controlled through transcriptional as well as post-transcriptional mechanisms. Both mechanisms involve activation of protein kinase A (PKA) into its subunits and subsequent phosphorylation and activation of several key regulatory factors. In rat C6 glioma cells, post-transcriptional gene regulation occurs through PKA mediated stabilization of LDH-A mRNA and subsequent increase of intracellular LDH A mRNA levels. Previous studies have demonstrated a cAMP-stabilizing region (CSR) located in the LDH-A 3'-untranslated region which, in combination with several phosphorylated CSR-binding proteins (CSR-BP), regulates the PKA-mediated stabilization of LDH-A mRNA. However, the mechanistic details of interaction of CSR with proteins as they pertain to mRNA stabilization by PKA are so far largely unknown. In this study we tested the hypothesis that ribosomal protein extracts (RSW) from glioma cells contain PKA regulatory (RII) and catalytic (C) subunits that, in combination with a protein kinase A anchoring protein (AKAP 95) and CSR BPs participate in forming CSR-protein complexes that are responsible for mRNA stability regulation. To demonstrate the importance of CSR-protein complex formation, the PKA subunits and AKAP 95 were removed from the RSW by immunoprecipitation, and the antigen-deleted RSW were subjected to CSR binding analysis using gel mobility shift and UV cross-linking. It was shown that AKAP 95 as well as RII formed a direct linkage with CSR during CSR-protein complex formation. In contrast, the catalytic subunit formed part of the CSR-protein complex but did not bind to CSR directly in a covalent linkage. To determine whether formation of CSR complexes that included C, RII, and AKAP 95 constituted a functional event and was necessary for mRNA stabilization, cell-free decay reactions were carried out with RSW extracts, and the kinetics of decay of LDH-A mRNA was determined. Depletion of PKA subunits and AKAP 95 from RSW extracts by immunoprecipitation resulted in a marked loss of mRNA stabilization activity indicating that the presence of the PKA regulatory and catalytic subunits as well as AKAP 95 in the CSR-protein complexes was absolutely necessary to achieve LDH-A mRNA stabilization. PMID- 15878852 TI - Rheb binding to mammalian target of rapamycin (mTOR) is regulated by amino acid sufficiency. AB - The removal of extracellular amino acids or leucine alone inhibits the ability of the mammalian target of rapamycin (mTOR) to signal to the raptor-dependent substrates, p70 S6 kinase and 4E-BP. This inhibition can be overcome by overexpression of the Rheb GTPase. Rheb binds directly to the amino-terminal lobe of the mTOR catalytic domain, and activates mTOR kinase in a GTP-dependent manner. Herein we show that the binding of Rheb to endogenous and recombinant mTOR is reversibly inhibited by withdrawal of all extracellular amino acids or just leucine. The effect of amino acid withdrawal is not attributable to changes in Rheb-GTP charging; amino acid withdrawal does not alter the GTP charging of recombinant Rheb. Moreover, the binding of mTOR to Rheb mutants that are unable to bind guanyl nucleotide in vivo is also inhibited by amino withdrawal. The inhibitory effect of amino acid withdrawal is exerted through an action on mTOR, at a site largely distinct from that responsible for the binding of Rheb; deletion of the larger, carboxyl-terminal lobe of the mTOR catalytic domain eliminates the inhibitory effect of amino acid withdrawal on Rheb binding, without altering Rheb binding per se. The lesser ability of the mTOR catalytic domain to bind Rheb after amino acid withdrawal does not persist after extraction and purification of the mTOR polypeptide. Amino acid withdrawal may generate an inhibitor of the Rheb-mTOR interaction that interferes with the signaling function of TOR complex 1. PMID- 15878853 TI - Analysis of the DNA substrate specificity of the human BACH1 helicase associated with breast cancer. AB - We have investigated the DNA substrate specificity of BACH1 (BRCA1-associated C terminal helicase). The importance of various DNA structural elements for efficient unwinding by purified recombinant BACH1 helicase was examined. The results indicated that BACH1 preferentially binds and unwinds a forked duplex substrate compared with a duplex flanked by only one single-stranded DNA (ssDNA) tail. In support of its DNA substrate preference, helicase sequestration studies revealed that BACH1 can be preferentially trapped by forked duplex molecules. BACH1 helicase requires a minimal 5 ' ssDNA tail of 15 nucleotides for unwinding of conventional duplex DNA substrates; however, the enzyme is able to catalytically release the third strand of the homologous recombination intermediate D-loop structure irrespective of DNA tail status. In contrast, BACH1 completely fails to unwind a synthetic Holliday junction structure. Moreover, BACH1 requires nucleic acid continuity in the 5 ' ssDNA tail of the forked duplex substrate within six nucleotides of the ssDNA-dsDNA junction to initiate efficiently DNA unwinding. These studies provide the first detailed information on the DNA substrate specificity of BACH1 helicase and provide insight to the types of DNA structures the enzyme is likely to act upon to perform its functions in DNA repair or recombination. PMID- 15878854 TI - Mammalian exocyst complex is required for the docking step of insulin vesicle exocytosis. AB - Glucose stimulates insulin secretion from pancreatic beta cells by inducing the recruitment and fusion of insulin vesicles to the plasma membrane. However, little is currently known about the mechanism of the initial docking or tethering of insulin vesicles prior to fusion. Here, we examined the role of the SEC6-SEC8 (exocyst) complex, implicated in trafficking of secretory vesicles to fusion sites in the plasma membrane in yeast and in regulating glucose-stimulated insulin secretion from pancreatic MIN6 beta cells. We show first that SEC6 is concentrated on insulin-positive vesicles, whereas SEC5 and SEC8 are largely confined to the cytoplasm and the plasma membrane, respectively. Overexpression of truncated, dominant-negative SEC8 or SEC10 mutants decreased the number of vesicles at the plasma membrane, whereas expression of truncated SEC6 or SEC8 inhibited overall insulin secretion. When single exocytotic events were imaged by total internal reflection fluorescence microscopy, the fluorescence of the insulin surrogate, neuropeptide Y-monomeric red fluorescent protein brightened, diffused, and then vanished with kinetics that were unaffected by overexpression of truncated SEC8 or SEC10. Together, these data suggest that the exocyst complex serves to selectively regulate the docking of insulin-containing vesicles at sites of release close to the plasma membrane. PMID- 15878855 TI - {beta}-Arrestin is crucial for ubiquitination and down-regulation of the insulin like growth factor-1 receptor by acting as adaptor for the MDM2 E3 ligase. AB - The insulin-like growth factor-1 receptor (IGF-1R) plays important roles in physiological growth and aging as well as promoting several crucial functions in cancer cells. However, the molecular mechanisms involved in expression and down regulation of IGF-1R are still poorly understood. Here we provide evidence that beta-arrestin, otherwise known to be involved in the regulation of G protein coupled receptors, serves as an adaptor to bring the oncoprotein E3 ubiquitin ligase MDM2 to the IGF-1R. In this way, beta-arrestin acts as a crucial component in the ubiquitination and down-regulation of the receptor. Both MDM2 and beta arrestin co-immunoprecipitated with the IGF-1R. The beta-arrestin isoform 1 appeared to be more strongly associated with the receptor than isoform 2, and in a molecular context it was 4-fold more efficient in inducing polyubiquitination of IGF-1R, a reaction that required the presence of beta-arrestin and MDM2. Ligand stimulation accelerated IGF-1R ubiquitination. In mouse P6 cells (overexpressing human IGF-1R) absence of beta-arrestin 1, but not of beta arrestin 2, blocked ubiquitination of IGF-1R. Conversely, in the two studied human melanoma cell lines both beta-arrestin isoforms seemed to be involved in IGF-1R ubiquitination. However, because depletion of beta-arrestin 1 almost completely eliminated degradation, and IGF-1 induced down-regulation of the receptor in these cells, whereas beta-arrestin 2 only had a partial effect, beta arrestin 1 seems to the more important isoform in affecting the expression of IGF 1R. To our knowledge this is the first study demonstrating a defined molecular role of beta-arrestin with direct relevance to cell growth and cancer. PMID- 15878856 TI - Anti-lipolytic action of AMP-activated protein kinase in rodent adipocytes. AB - Despite its importance in terms of energy homeostasis, the role of AMP-activated protein kinase in adipose tissue remains controversial. Initial studies have described an anti-lipolytic role for AMP-activated protein kinase, whereas more recent studies have suggested the converse. Thus we have addressed the role of AMP-activated protein kinase in adipose tissue by modulating AMP-activated protein kinase activity in primary rodent adipocytes using pharmacological activators or by adenoviral expression of dominant negative or constitutively active forms of the kinase. We then studied the effects of AMP-activated protein kinase activity modulation on lipolytic mechanisms. Finally, we analyzed the consequences of a genetic deletion of AMP-activated protein kinase in mouse adipocytes. AMP-activated protein kinase activity in adipocytes is represented mainly by the alpha(1) isoform and is induced by all of the stimuli that increase cAMP in adipocytes, including fasting. When AMP-activated protein kinase activity is increased by 5-aminoimidazole-4-carboxamide-riboside, phenformin, or by the expression of a constitutively active form, isoproterenol-induced lipolysis is strongly reduced. Conversely, when AMP-activated protein kinase activity is decreased either by a dominant negative form or in AMP-activated protein kinase alpha(1) knock-out mice, lipolysis is increased. We present data suggesting that AMP-activated protein kinase acts on hormone-sensitive lipase by blocking its translocation to the lipid droplet. We conclude that, in mature adipocytes, AMP activated protein kinase activation has a clear anti-lipolytic effect. PMID- 15878857 TI - Identification and active expression of the Mycobacterium tuberculosis gene encoding 5-phospho-{alpha}-d-ribose-1-diphosphate: decaprenyl-phosphate 5 phosphoribosyltransferase, the first enzyme committed to decaprenylphosphoryl-d arabinose synthesis. AB - Decaprenylphosphoryl-d-arabinose, the lipid donor of mycobacterial d arabinofuranosyl residues, is synthesized from phosphoribose diphosphate rather than from a sugar nucleotide. The first committed step in the process is the transfer of a 5-phosphoribosyl residue from phosphoribose diphosphate to decaprenyl phosphate to form decaprenylphosphoryl-5-phosphoribose via a 5-phospho alpha-d-ribose-1-diphosphate:decaprenyl-phosphate 5-phospho-ribosyltransferase. A candidate for the gene encoding this enzyme (Rv3806c) was identified in Mycobacterium tuberculosis, primarily via its homology to one of four genes responsible for d-arabinosylation of nodulation factor in Azorhizobium caulinodans. The resulting protein was predicted to contain eight or nine transmembrane domains. The gene was expressed in Escherichia coli, and membranes from the expression strain of E. coli but not from a control strain of E. coli were shown to convert phosphoribose diphosphate and decaprenyl phosphate into decaprenylphosphoryl-5-phosphoribose. Neither UDP-galactose nor GDP-mannose was active as a sugar donor. The enzyme favored polyprenyl phosphate with 50-60 carbon atoms, was unable to use C-20 polyprenyl phosphate, and used C-75 polyprenyl phosphate less efficiently than C-50 or C-60. It requires CHAPS detergent and Mg(2+) for activity. The Rv3806c gene encoding 5-phospho-alpha-d ribose-1-diphosphate:decaprenyl-phosphate 5-phosphoribosyltransferase is known to be essential for the growth of M. tuberculosis, and the tuberculosis drug ethambutol inhibits other steps in arabinan biosynthesis. Thus the Rv3806c encoded enzyme appears to be a good target for the development of new tuberculosis drugs. PMID- 15878858 TI - The iron-sulfur cluster of the Rieske iron-sulfur protein functions as a proton exiting gate in the cytochrome bc(1) complex. AB - The destruction of the Rieske iron-sulfur cluster ([2Fe-2S]) in the bc(1) complex by hematoporphyrin-promoted photoinactivation resulted in the complex becoming proton-permeable. To study further the role of this [2Fe-2S] cluster in proton translocation of the bc(1) complex, Rhodobacter sphaeroides mutants expressing His-tagged cytochrome bc(1) complexes with mutations at the histidine ligands of the [2Fe-2S] cluster were generated and characterized. These mutants lacked the [2Fe-2S] cluster and possessed no bc(1) activity. When the mutant complex was co inlaid in phospholipid vesicles with intact bovine mitochondrial bc(1) complex or cytochrome c oxidase, the proton ejection, normally observed in intact reductase or oxidase vesicles during the oxidation of their corresponding substrates, disappeared. This indicated the creation of a proton-leaking channel in the mutant complex, whose [2Fe-2S] cluster was lacking. Insertion of the bc(1) complex lacking the head domain of the Rieske iron-sulfur protein, removed by thermolysin digestion, into PL vesicles together with mitochondrial bc(1) complex also rendered the vesicles proton-permeable. Addition of the excess purified head domain of the Rieske iron-sulfur protein partially restored the proton-pumping activity. These results indicated that elimination of the [2Fe-2S] cluster in mutant bc(1) complexes opened up an otherwise closed proton channel within the bc(1) complex. It was speculated that in the normal catalytic cycle of the bc(1) complex, the [2Fe-2S] cluster may function as a proton-exiting gate. PMID- 15878859 TI - Crystallin {gamma}B-I4F mutant protein binds to {alpha}-crystallin and affects lens transparency. AB - A new mouse mutant line, Clapper, identified from N-ethyl-N-nitrosurea (ENU) mutagenized mice, develops a dominant lamellar cataract. The cataract blocks the image of retinal fundus and transmits a fuzzy fluorescein image of retinal vasculature during angiography. The cataractous lens opacity decreases as the mice age. The Clapper mutation has been identified to be a missense mutation of the gammaB-crystallin gene that replaces the 4th isoleucine residue with a phenylalanine (gammaB-I4F). Unlike wild type gammaB, the gammaB-I4F mutant protein binds to alpha-crystallin to form high molecular weight complexes in vivo and in vitro. Circular dichroism measurements indicate that gammaB-I4F protein is less stable than wild type gammaB at high temperature. Darkly stained aggregates, enlarged interfiber spaces, and disorganized and smaller inner mature fibers were found in the regions of the cataract in homozygous Clapper mutant lenses. Thus, the lamellar cataract is likely due to the light-scattering effects of the enlarged interfiber spaces and protein aggregates caused by gammaB-I4F mutant proteins interacting with alpha-crystallin in the lens. PMID- 15878860 TI - Mechanism of the conversion of xanthine dehydrogenase to xanthine oxidase: identification of the two cysteine disulfide bonds and crystal structure of a non convertible rat liver xanthine dehydrogenase mutant. AB - Mammalian xanthine dehydrogenase can be converted to xanthine oxidase by modification of cysteine residues or by proteolysis of the enzyme polypeptide chain. Here we present evidence that the Cys(535) and Cys(992) residues of rat liver enzyme are indeed involved in the rapid conversion from the dehydrogenase to the oxidase. The purified mutants C535A and/or C992R were significantly resistant to conversion by incubation with 4,4'-dithiodipyridine, whereas the recombinant wild-type enzyme converted readily to the oxidase type, indicating that these residues are responsible for the rapid conversion. The C535A/C992R mutant, however, converted very slowly during prolonged incubation with 4,4' dithiodipyridine, and this slow conversion was blocked by the addition of NADH, suggesting that another cysteine couple located near the NAD(+) binding site is responsible for the slower conversion. On the other hand, the C535A/C992R/C1316S and C535A/C992R/C1324S mutants were completely resistant to conversion, even on prolonged incubation with 4,4'-dithiodipyridine, indicating that Cys(1316) and Cys(1324) are responsible for the slow conversion. The crystal structure of the C535A/C992R/C1324S mutant was determined in its demolybdo form, confirming its dehydrogenase conformation. PMID- 15878861 TI - Activated mitofusin 2 signals mitochondrial fusion, interferes with Bax activation, and reduces susceptibility to radical induced depolarization. AB - Mitochondrial fusion in higher eukaryotes requires at least two essential GTPases, Mitofusin 1 and Mitofusin 2 (Mfn2). We have created an activated mutant of Mfn2, which shows increased rates of nucleotide exchange and decreased rates of hydrolysis relative to wild type Mfn2. Mitochondrial fusion is stimulated dramatically within heterokaryons expressing this mutant, demonstrating that hydrolysis is not requisite for the fusion event, and supporting a role for Mfn2 as a signaling GTPase. Although steady-state mitochondrial fusion required the conserved intermembrane space tryptophan residue, this requirement was overcome within the context of the hydrolysis-deficient mutant. Furthermore, the punctate localization of Mfn2 is lost in the dominant active mutants, indicating that these sites are functionally controlled by changes in the nucleotide state of Mfn2. Upon staurosporine-stimulated cell death, activated Bax is recruited to the Mfn2-containing puncta; however, Bax activation and cytochrome c release are inhibited in the presence of the dominant active mutants of Mfn2. The dominant active form of Mfn2 also protected the mitochondria against free radical-induced permeability transition. In contrast to staurosporine-induced outer membrane permeability transition, pore opening induced through the introduction of free radicals was dependent upon the conserved intermembrane space residue. This is the first evidence that Mfn2 is a signaling GTPase regulating mitochondrial fusion and that the nucleotide-dependent activation of Mfn2 concomitantly protects the organelle from permeability transition. The data provide new insights into the critical relationship between mitochondrial membrane dynamics and programmed cell death. PMID- 15878862 TI - Fine tuning of the specificity of an anti-progesterone antibody by first and second sphere residue engineering. AB - The specificity of anti-progesterone P15G12C12G11 antibody was improved by combination of in vitro scanning saturation mutagenesis and error-prone PCR. The most evolved mutant is able to discriminate against 5beta- or 5alpha dihydroprogesterone, 23 and 15 times better than the starting antibody, while maintaining the affinity for progesterone that remains in the picomolar range. The high level of homology with anti-progesterone monoclonal antibody DB3 allowed the construction of three-dimensional models of P15G12C12G11 based on the structures of DB3 in complex with various steroids. These models together with binding data, derived from site-directed mutagenesis, were used to build a phage library in which five first sphere positions in complementarity-determining regions 2H and 3L were varied. Variants selected by an initial screening in competition against a large excess of 5beta- or 5alpha-dihydroprogesterone were characterized by a convergent amino acid signature different from that of the wild-type antibody and had lower cross-reactivity. Binding properties of this first set of mutants were further improved by the addition of second sphere mutations selected independently from an error-prone library. The three dimensional models of the best variant show changes in the antigen binding site that explain well the increase in selectivity. The improvements are partly linked to a change in the canonical class of the light chain third hypervariable loop. PMID- 15878863 TI - A role for mixed lineage kinases in regulating transcription factor CCAAT/enhancer-binding protein-{beta}-dependent gene expression in response to interferon-{gamma}. AB - Transcription factor CCAAT/enhancer-binding protein-beta (C/EBP-beta) regulates a variety of cellular functions in response to exogenous stimuli. We have reported earlier that C/EBP-beta induces gene transcription through a novel interferon (IFN)-response element called gamma-IFN-activated transcriptional element. We show here that IFN-gamma-induced, C/EBP-beta/gamma-IFN-activated transcriptional element-dependent gene expression is regulated by mixed lineage kinases (MLKs), members of the mitogen-activated protein kinase kinase kinase family. MLK3 appears to activate C/EBP-beta in response to IFN-gamma by a mechanism involving decreased phosphorylation of a specific phosphoacceptor residue, Ser(64), within the transactivation domain. Decreased phosphorylation of Ser(64) was independent of IFN-gamma-stimulated ERK1/2 activation and did not require the ERK phosphorylation site Thr(189) located in regulatory domain 2 of C/EBP-beta. Together these studies provide the first evidence that MLK3 is involved in IFN gamma signaling and identify a novel mechanism of transcriptional activation by IFN-gamma. PMID- 15878864 TI - Kaposi's sarcoma-associated herpesvirus activation of vascular endothelial growth factor receptor 3 alters endothelial function and enhances infection. AB - Kaposi's sarcoma-associated herpesvirus (KSHV; also known as human herpesvirus 8) is the etiologic agent of Kaposi's sarcoma, an endothelial neoplasm. This gamma herpesvirus encodes for several unique proteins that alter target cell function, including the virion envelope-associated glycoprotein B (gB). Glycoprotein B has an RGD (Arg-Gly-Asp) motif at the extracellular amino terminus region and binds to the alpha3beta1 surface integrin, which enhances virus entry. We now report that gB can activate the vascular endothelial growth factor receptor 3 (VEGFR-3) on the surface of microvascular endothelial cells and trigger receptor signaling, which can modulate endothelial migration and proliferation. Furthermore, we observed that VEGFR-3 expression and activation enhance KSHV infection and participate in KSHV-mediated transformation. These functional changes in the endothelium may contribute to the pathogenesis of Kaposi's sarcoma and suggest that interventions that inhibit gB activation of VEGFR-3 could be useful in the treatment of this neoplasm. PMID- 15878865 TI - Statins exert endothelial atheroprotective effects via the KLF2 transcription factor. AB - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, statins, have been shown to positively impact vascular function independent of their plasma lipid lowering action. Several of these beneficial effects involve modulation of gene expression. Here we explored whether the transcription factor Kruppel-like factor 2 (KLF2), a biomechanically activated gene we recently identified as part of the endothelial "atheroprotective phenotype," is regulated by statins and whether this mechanism is important for the non-lipid lowering beneficial effects mediated by these drugs in endothelium. The mRNA levels of KLF2 in human umbilical vein endothelial cells increased in the presence of various statins. KLF2 induction was observed within 8 h after drug treatment and remained elevated for at least 24 h. This statin effect on KLF2 expression was reversed by addition of mevalonate and its downstream metabolite geranygeranyl pyrophosphate. Furthermore, inhibition of protein geranylgeranylation with GGTI-298 significantly induced KLF2 levels, whereas inhibition of farnesylation did not. Statin-mediated KLF2 expression was followed by the up-regulation of several of its downstream transcriptional targets. Using small interfering RNA to block KLF2 expression, we demonstrated that this transcription factor is necessary for the statin-mediated regulation of several pathophysiologically relevant genes. These results strongly implicate KLF2 as a transcriptional regulator of the statin mediated effects in vascular endothelium and provide a novel mechanism for the well established non-lipid lowering beneficial cardiovascular effects of statins. PMID- 15878866 TI - Role of Tim21 in mitochondrial translocation contact sites. AB - Translocation of preproteins with N-terminal presequences into mitochondria requires the cooperation of the translocase of the outer membrane (TOM complex) and the presequence translocase of the inner membrane (TIM23 complex). However, the molecular nature of the translocation contact sites is poorly understood. We have identified a novel component of the TIM23 translocase, Tim21, which is involved in their formation. Tim21 is anchored in the mitochondrial inner membrane by a single transmembrane domain and exposes its C-terminal domain into the intermembrane space. The purified C-terminal domain of Tim21 appears not to bind to any of the TIM23 components but rather specifically interacts with the TOM complex. We propose that Tim21 binds to the trans site of the TOM complex thus keeping the two translocases in close contact. PMID- 15878867 TI - Alpha-tocopheryl succinate inhibits malignant mesothelioma by disrupting the fibroblast growth factor autocrine loop: mechanism and the role of oxidative stress. AB - We have studied the potential effect against human malignant mesotheliomas (MM) of alpha-tocopheryl succinate (alpha-TOS), a redox-silent vitamin E analog with strong pro-apoptotic and anti-cancer activity. alpha-TOS at sub-apoptotic levels inhibited proliferation of MM cell lines, while being nontoxic to nonmalignant mesothelial cells. Because MM cells are typified by a highly metastatic phenotype, we investigated the effect of alpha-TOS on genes playing a major role in MM progression. Of these, alpha-TOS down regulated fibroblast growth factor (FGF)-1 and, in particular, FGF-2 on the transcriptional level in MM cells, and this was not observed in their nonmalignant counterparts. FGF-2 short interfering RNA suppressed proliferation of MM cells. Down-regulation of FGF-2 was likely because of inhibition of the egr-1 transcription activity that was decreased in MM cells via oxidative stress induced by alpha-TOS, as evidenced by EPR spectroscopy, whereas nonmalignant cells did not show this response. Treatment of MM cells with egr-1 short interfering RNA suppressed proliferation, which was overridden by exogenously added recombinant FGF-1 and, in particular, FGF-2. An analog of coenzyme Q targeted to mitochondria and superoxide dismutase overrode inhibition of MM cell proliferation by alpha-TOS as well as alpha-TOS-induced inhibition of egr-1-dependent transactivation. Finally, alpha-TOS significantly suppressed experimental MM in immunocompromised mice. Our data suggest that alpha TOS suppresses MM cell proliferation by disrupting the FGF-FGF receptor autocrine signaling loop by generating oxidative stress and point to the agent as a selective drug against thus far fatal mesotheliomas. PMID- 15878868 TI - Eukaryotic translation initiation factor 4E is a cellular target for toxicity and death due to exposure to cadmium chloride. AB - Whether translation initiation factor 4E (eIF4E), the mRNA cap binding and rate limiting factor required for translation, is a target for cytotoxicity and cell death induced by cadmium, a human carcinogen, was investigated. Exposure of human cell lines, HCT15, PLC/PR/5, HeLa, and Chang, to cadmium chloride resulted in cytotoxicity and cell death, and this was associated with a significant decrease in eIF4E protein levels. Similarly, specific silencing of the expression of the eIF4E gene, caused by a small interfering RNA, resulted in significant cytotoxicity and cell death. On the other hand, overexpression of the eIF4E gene was protective against the cadmium-induced cytotoxicity and cell death. Further studies revealed the absence of alterations in the eIF4E mRNA level in the cadmium-treated cells despite their decreased eIF4E protein level. In addition, exposure of cells to cadmium resulted in enhanced ubiquitination of eIF4E protein while inhibitors of proteasome activity reversed the cadmium-induced decrease of eIF4E protein. Exposure of cells to cadmium, as well as the specific silencing of eIF4E gene, also resulted in decreased cellular levels of cyclin D1, a critical cell cycle and growth regulating gene, suggesting that the observed inhibition of cyclin D1 gene expression in the cadmium-treated cells is most likely due to decreased cellular level of eIF4E. Taken together, our results demonstrate that the exposure of cells to cadmium chloride resulted in cytotoxicity and cell death due to enhanced ubiquitination and consequent proteolysis of eIF4E protein, which in turn diminished cellular levels of critical genes such as cyclin D1. PMID- 15878869 TI - Membrane type-1 matrix metalloproteinase (MT1-MMP) exhibits an important intracellular cleavage function and causes chromosome instability. AB - Elevated expression of membrane type-1 matrix metalloproteinase (MT1-MMP) is closely associated with malignancies. There is a consensus among scientists that cell surface-associated MT1-MMP is a key player in pericellular proteolytic events. Now we have identified an intracellular, hitherto unknown, function of MT1-MMP. We demonstrated that MT1-MMP is trafficked along the tubulin cytoskeleton. A fraction of cellular MT1-MMP accumulates in the centrosomal compartment. MT1-MMP targets an integral centrosomal protein, pericentrin. Pericentrin is known to be essential to the normal functioning of centrosomes and to mitotic spindle formation. Expression of MT1-MMP stimulates mitotic spindle aberrations and aneuploidy in non-malignant cells. Volumes of data indicate that chromosome instability is an early event of carcinogenesis. In agreement, the presence of MT1-MMP activity correlates with degraded pericentrin in tumor biopsies, whereas normal tissues exhibit intact pericentrin. We believe that our data show a novel proteolytic pathway to chromatin instability and elucidate the close association of MT1-MMP with malignant transformation. PMID- 15878870 TI - Functional selectivity of G protein signaling by agonist peptides and thrombin for the protease-activated receptor-1. AB - Thrombin activates protease-activated receptor-1 (PAR-1) by cleavage of the amino terminus to unmask a tethered ligand. Although peptide analogs can activate PAR 1, we show that the functional responses mediated via PAR-1 differ between the agonists. Thrombin caused endothelial monolayer permeability and mobilized intracellular calcium with EC(50) values of 0.1 and 1.7 nm, respectively. The opposite order of activation was observed for agonist peptide (SFLLRN-CONH(2) or TFLLRNKPDK) activation. The addition of inactivated thrombin did not affect agonist peptide signaling, suggesting that the differences in activation mechanisms are intramolecular in origin. Although activation of PAR-1 or PAR-2 by agonist peptides induced calcium mobilization, only PAR-1 activation affected barrier function. Induced barrier permeability is likely to be Galpha(12/13) mediated as chelation of Galpha(q)-mediated intracellular calcium with BAPTA-AM, pertussis toxin inhibition of Galpha(i/o), or GM6001 inhibition of matrix metalloproteinase had no effect, whereas Y-27632 inhibition of the Galpha(12/13) mediated Rho kinase abrogated the response. Similarly, calcium mobilization is Galpha(q)-mediated and independent of Galpha(i/o) and Galpha(12/13) because pertussis toxin Y-27632 and had no effect, whereas U-73122 inhibition of phospholipase C-beta blocked the response. It is therefore likely that changes in permeability reflect Galpha(12/13) activation, and changes in calcium reflect Galpha(q) activation, implying that the pharmacological differences between agonists are likely caused by the ability of the receptor to activate Galpha(12/13) or Galpha(q). This functional selectivity was characterized quantitatively by a mathematical model describing each step leading to Rho activation and/or calcium mobilization. This model provides an estimate that peptide activation alters receptor/G protein binding to favor Galpha(q) activation over Galpha(12/13) by approximately 800-fold. PMID- 15878871 TI - Analysis of binding sites in human C-reactive protein for Fc{gamma}RI, Fc{gamma}RIIA, and C1q by site-directed mutagenesis. AB - Human C-reactive protein (CRP) is a classical, acute phase serum protein synthesized by the liver in response to infection, inflammation, or trauma. CRP binds to microbial antigens and damaged cells, opsonizes particles for phagocytosis and regulates the inflammatory response by the induction of cytokine synthesis. These activities of CRP depend on its ability to activate complement and to bind to Fcgamma receptors (FcgammaR). The goal of this study was to elucidate amino acid residues important for the interaction of CRP with human FcgammaRI (CD64) and FcgammaRIIa (CD32). Several mutations of the CRP structure were studied based on the published crystal structure of CRP. Mutant and wild type recombinant CRP molecules were expressed in the baculovirus system and their interactions with FcgammaR and C1q were determined. A previous study by our laboratory identified an amino acid position, Leu(176), critical for CRP binding to FcgammaRI and work by others (Agrawal, A., Shrive, A. K., Greenhough, T. J., and Volanakis, J. E. (2001) J. Immunol. 166, 3998-4004) determined several residues important for C1q binding. The amino acid residues important to CRP binding to FcgammaRIIa were previously unknown. This study newly identifies residues Thr(173) and Asn(186) as important for the binding of CRP to FcgammaRIIa and FcgammaRI. Lys(114), like Leu(176), was implicated in binding to FcgammaRI, but not FcgammaRIIa. Single mutations at amino acid positions Lys(114), Asp(169), Thr(173), Tyr(175), and Leu(176) affected C1q binding to CRP. These results further identify amino acids involved in the binding sites on CRP for FcgammaRI, FcgammaRIIa, and C1q and indicate that these sites are overlapping. PMID- 15878872 TI - Sterically locked synthetic bilin derivatives and phytochrome Agp1 from Agrobacterium tumefaciens form photoinsensitive Pr- and Pfr-like adducts. AB - Phytochrome photoreceptors undergo reversible photoconversion between the red absorbing form, Pr, and the far-red-absorbing form, Pfr. The first step in the conversion from Pr to Pfr is a Z to E isomerization around the C15=C16 double bond of the bilin chromophore. We prepared four synthetic biliverdin (BV) derivatives in which rings C and D are sterically locked by cyclizing with an additional carbon chain. In these chromophores, which are termed 15Za, 15Zs, 15Ea, and 15Es, the C15=C16 double bond is in either the Z or E configuration and the C14-C15 single bond in either the syn or anti conformation. The chromophores were assembled with Agrobacterium phytochrome Agp1, which incorporates BV as natural chromophore. All locked BV derivatives bound covalently to the protein and formed adducts with characteristic spectral properties. The 15Za adduct was spectrally similar to the Pr form and the 15Ea adduct similar to the Pfr form of the BV adduct. Thus, the chromophore of Agp1 adopts a C15=C16 Z configuration and a C14-C15 anti conformation in the Pr form and a C15=C16 E configuration and a C14-C15 anti conformation in the Pfr form. Both the 15Zs and the 15Es adducts absorbed only in the blue region of the visible spectra. All chromophore adducts were analyzed by size exclusion chromatography and histidine kinase activity to probe for protein conformation. In either case, the 15Za adduct behaved like the Pr and the 15Ea adduct like the Pfr form of Agp1. Replacing the natural chromophore by a locked 15Ea derivative can thus bring phytochrome holoprotein in the Pfr form in darkness. In this way, physiological action of Pfr can be studied in vivo and separated from Pr/Pfr cycling and other light effects. PMID- 15878873 TI - A cryptic Rab1-binding site in the p115 tethering protein. AB - Small GTPases and coiled-coil proteins of the golgin family help to tether COPI vesicles to Golgi membranes. At the cis-side of the Golgi, the Rab1 GTPase binds directly to each of three coiled-coil proteins: p115, GM130, and as now shown, Giantin. Rab1 binds to a coiled-coil region within the tail domain of p115 and this binding is inhibited by the C-terminal, acidic domain of p115. Furthermore, GM130 and Giantin bind to the acidic domain of p115 and stimulate p115 binding to Rab1, suggesting that p115 binding to Rab1 is regulated. Regulation of this interaction by proteins such as GM130 and Giantin may control the membrane recruitment of p115 by Rab1. PMID- 15878874 TI - Mitochondrial glycerol-3-phosphate acyltransferase-1 is essential in liver for the metabolism of excess acyl-CoAs. AB - In vitro studies suggest that the mitochondrial glycerol-3-phosphate acyltransferase-1 (mtGPAT1) isoform catalyzes the initial and rate-controlling step in glycerolipid synthesis and aids in partitioning acyl-CoAs toward triacylglycerol synthesis and away from degradative pathways. To determine whether the absence of mtGPAT1 would increase oxidation of acyl-CoAs and restrict the development of hepatic steatosis, we fed wild type and mtGPAT1-/- mice a diet high in fat and sucrose (HH) for 4 months to induce the development of obesity and a fatty liver. Control mice were fed a diet low in fat and sucrose (LL). With the HH diet, absence of mtGPAT1 resulted in increased partitioning of acyl-CoAs toward oxidative pathways, demonstrated by 60% lower hepatic triacylglycerol content and 2-fold increases in plasma beta-hydroxybutyrate, acylcarnitines, and hepatic mRNA expression of mitochondrial HMG-CoA synthase. Despite the increase in fatty acid oxidation, liver acyl-CoA levels were 3-fold higher in the mtGPAT1 /- mice fed both diets. A lack of difference in CPT1 and FAS mRNA expression between genotypes suggested that the increased acyl-CoA content was not because of increased de novo synthesis, but instead, to an impaired ability to use long chain acyl-CoAs derived from the diet, even when the dietary fat content was low. Hyperinsulinemia and reduced glucose tolerance on the HH diet was greater in the mtGPAT1-/- mice, which did not suppress the expression of the gluconeogenic genes glucose-6-phosphatase and phosphoenolpyruvate carboxykinase. This study demonstrates that mtGPAT1 is essential for normal acyl-CoA metabolism, and that the absence of hepatic mtGPAT1 results in the partitioning of fatty acids away from triacylglycerol synthesis and toward oxidation and ketogenesis. PMID- 15878875 TI - Caenorhabditus elegans arrestin regulates neural G protein signaling and olfactory adaptation and recovery. AB - Although regulation of G protein-coupled receptor signaling by receptor kinases and arrestins is a well established biochemical process, the physiological significance of such regulation remains poorly understood. To better understand the in vivo consequences of arrestin function, we have examined the function of the sole arrestin in Caenorhabditis elegans (ARR-1). ARR-1 is primarily expressed in the nervous system, including the HSN neuron and various chemosensory neurons involved in detecting soluble and volatile odorants. arr-1 null mutants exhibit normal chemotaxis but have significant defects in olfactory adaptation and recovery to volatile odorants. In contrast, adaptation is enhanced in animals overexpressing ARR-1. Both the adaptation and recovery defects of arr-1 mutants are rescued by transgenic expression of wild-type ARR-1, whereas expression of a C-terminally truncated ARR-1 effectively rescues only the adaptation defect. A potential mechanistic basis for these findings is revealed by in vitro studies demonstrating that wild-type ARR-1 binds proteins of the endocytic machinery and promotes receptor endocytosis, whereas C-terminally truncated ARR-1 does not. These results demonstrate that ARR-1 functions to regulate chemosensory signaling, enabling organisms to adapt to a variety of environmental cues, and provide an in vivo link between arrestin, receptor endocytosis, and temporal recovery from adaptation. PMID- 15878876 TI - Transcription-induced chromatin remodeling at the c-myc gene involves the local exchange of histone H2A.Z. AB - The post-translational modification of histones and the incorporation of core histone variants play key roles in governing gene expression. Many eukaryotic genes regulate their expression by limiting the escape of RNA polymerase from promoter-proximal pause sites. Here we report that elongating RNA polymerase II complexes encounter distinct chromatin landscapes that are marked by methylation of lysine residues Lys(4), Lys(79), and Lys(36) of histone H3. However, neither histone methylation nor acetylation directly regulates the release of elongation complexes stalled at promoter-proximal pause sites of the c-myc gene. In contrast, transcriptional activation is associated with local displacement of the histone variant H2A.Z within the transcribed region and incorporation of the major histone variant H2A. This result indicates that transcribing RNA polymerase II remodels chromatin in part through coincident displacement of H2A.Z-H2B dimers and incorporation of H2A-H2B dimers. In combination, these results suggest a new model in which the incorporation of H2A.Z into nucleosomes down-regulates transcription; at the same time it may act as a cellular memory for transcriptionally poised gene domains. PMID- 15878877 TI - Apolipoprotein A-V-heparin interactions: implications for plasma lipoprotein metabolism. AB - Transgenic and gene disruption experiments in mice have revealed that apolipoprotein (apo) A-V is a potent regulator of plasma triglyceride (TG) levels. To investigate the molecular basis of apoA-V function, the ability of isolated recombinant apoA-V to modulate lipoprotein lipase (LPL) activity was examined in vitro. With three distinct lipid substrate particles, including very low-density lipoprotein (VLDL), a TG/phospholipid emulsion, or dimyristoylphosphatidylcholine liposomes, apoA-V had little effect on LPL activity. In the absence or presence apolipoprotein C-II, apoA-V marginally inhibited LPL activity. On the other hand, apoA-V-dimyristoylphosphatidylcholine disc particles bound to heparin-Sepharose and were specifically eluted upon application of a linear gradient of NaCl. The interaction of apoA-V with sulfated glycosaminoglycans was further studied by surface plasmon resonance spectroscopy. ApoA-V showed strong binding to heparin-coated chips, and binding was competed by free heparin. ApoA-V enrichment enhanced binding of apoC-II-deficient chylomicrons and VLDL to heparin-coated chips. When LPL was first bound to the heparin-coated chip, apoA-V-enriched chylomicrons showed binding. Finally, human pre- and post-heparin plasma samples were subjected to immunoblot analysis with anti-apoA-V IgG. No differences in the amount of apoA-V present were detected. Taken together, the results show that apoA-V lipid complexes bind heparin and, when present on TG-rich lipoprotein particles, may promote their association with cell surface heparan sulfate proteoglycans. Through such interactions, apoA-V may indirectly affect LPL activity, possibly explaining its inverse correlation with plasma TG levels. PMID- 15878878 TI - Mechanisms of cytoplasmic {beta}-catenin accumulation and its involvement in tumorigenic activities mediated by oncogenic splicing variant of the receptor originated from Nantes tyrosine kinase. AB - The beta-catenin pathway plays a critical role in the pathogenesis of certain types of cancers. To gain insight into mechanisms by which altered receptor tyrosine kinases regulate cytoplasmic beta-catenin accumulation, the effect of an oncogenic receptor originated from Nantes (RON) variant on beta-catenin accumulation and the role of beta-catenin in RON-mediated tumorigenic activities were studied. In NIH3T3 cells harboring oncogenic variant RONDelta160, increased beta-catenin accumulation with tyrosine phosphorylation and nuclear translocation was observed. Overexpression of RONDelta160 also resulted in increased expression of beta-catenin target genes c-myc and cyclin D1. By analyzing cellular proteins that regulate beta-catenin stabilities, it was found that RONDelta160 activates the protein disheveled (DVL) and inactivates glycogen synthase kinase-3beta by Ser-9 residue phosphorylation. These effects were channeled by RONDelta160 activated PI 3-kinase-AKT pathways that are sensitive to specific inhibitors, such as wortmannin, but not to other chemical inhibitors. Silencing RONDelta160 expression by specific small interfering RNA blocked not only beta-catenin expression but also c-myc and cyclin D1 expression, suggesting that RON expression is required for the activation of the beta-catenin signaling pathway. Moreover, it was found that knockdown of the beta-catenin gene expression by small interfering RNA techniques reduces significantly the RONDelta160-mediated NIH3T3 cell proliferation, focus-forming activities and anchorage-independent growth. Thus, the oncogenic RON variant regulates beta-catenin stabilities through activation of DVL and inactivation of glycogen synthase kinase-3beta. The activated beta-catenin cascade is one of the pathways involved in tumorigenic activities mediated by the oncogenic RON variant. PMID- 15878879 TI - Alkali cation binding and permeation in the rat organic cation transporter rOCT2. AB - Organic cation transporters of the OCT family mediate downhill transport of organic cations, compatible with carrier, pore, or gate-lumen-gate mechanisms. We studied rat OCT2 expressed in Xenopus oocytes by the two-electrode voltage-clamp technique, including membrane capacitance (C(m)) monitoring. Choline, a transported cationic substrate, elicited the expected inward currents but also elicited decreases of C(m). Similar C(m) decreases were caused by the non transported inhibitors tetrabutylammonium (a cation) and corticosterone (uncharged). Effects on C(m) were voltage-dependent, with a maximum at -140 mV. These findings suggest that the empty rOCT2 protein can undergo an electrogenic conformation change, with one conformation highly favored at physiological voltage. Moreover, alkali cations elicited considerable inward currents and inhibited uptake of [(14)C]tetraethylammonium with a sequence Cs(+) > Rb(+) > K(+) > Na(+) approximately Li(+). Cs(+) affected current and capacitance with similar affinity (K(0.5) approximately 50 mm). Tetraethylammonium inhibited Cs(+) currents in a concentration-dependent manner. Conversely, Cs(+) inhibited tetraethylammonium uptake by a competitive mechanism. Activation energy of the currents estimated from measurements between 12 degrees C and 32 degrees C was approximately 81 kJ/mol for Cs(+) and 39 kJ/mol for tetramethylammonium, compatible with permeation of Cs(+) through rOCT2 along the same path as organic substrates and by a mechanism different from simple electrodiffusion. Rationalization of Cs(+) selectivity in terms of a pore pointed to a pore diameter of approximately 4 A. Intriguingly, that value matches the known selectivity of rOCT2 for organic compounds. Our data show that selective permeability of rOCT2 is not determined by ligand affinity but might rather be understood in terms of the ion channel concept of a distinct "selectivity filter." PMID- 15878880 TI - Transcriptional activity of Sp1 is regulated by molecular interactions between the zinc finger DNA binding domain and the inhibitory domain with corepressors, and this interaction is modulated by MEK. AB - Sp1 activates the transcription of many cellular and viral genes with the GC-box in either the proximal promoter or the enhancer. Sp1 is composed of several functional domains, such as the inhibitory domain (ID), two serine/threonine-rich domains, two glutamine-rich domains, three C2H2-type zinc finger DNA binding domains (ZFDBD), and a C-terminal D domain. The ZDDBD is the most highly conserved domain among the Sp-family transcription factors and plays a critical role in GC-box recognition. In this study, we investigated the protein-protein interactions occurring at the Sp1ZFDBD and the Sp1ID, and the molecular mechanisms controlling the interaction. Our results found that Sp1ZFDBD and Sp1ID repressed transcription once they were targeted to the proximal promoter of the pGal4 UAS reporter fusion gene system, suggesting molecular interaction with the repressor molecules. Indeed, mammalian two-hybrid assays, GST fusion protein pull down assays, and co-immunoprecipitation assays showed that Sp1ZFDBD and Sp1ID are able to interact with corepressor proteins such as SMRT, NcoR, and BCoR. The molecular interactions appear to be regulated by MAP kinase/Erk kinase kinase (MEK). The molecular interactions between Sp1ID and the corepressor might explain the role of Sp1 as a repressor under certain circumstances. The siRNA-induced degradation of the corepressors resulted in an up-regulation of Sp1-dependent transcription. The cellular context of the corepressors and the regulation of molecular interaction between corepressors and Sp1ZFDBD or Sp1ID might be important in controlling Sp1 activity. PMID- 15878881 TI - Comprehensive analysis of cytosolic Nudix hydrolases in Arabidopsis thaliana. AB - Nudix hydrolases are a family of proteins that catalyze the hydrolysis of a variety of nucleoside diphosphate derivatives. Twenty-four genes of the Nudix hydrolase homologues (AtNUDTs) with predicted localizations in the cytosol, chloroplasts, and mitochondria exist in Arabidopsis thaliana. Here, we demonstrated the comprehensive analysis of nine types of cytosolic AtNUDT proteins (AtNUDT1, -2, -4, -5, -6, -7, -9, -10, and -11). The recombinant proteins of AtNUDT2, -6, -7, and -10 showed both ADP-ribose and NADH pyrophosphatase activities with significantly high affinities compared with those of animal and yeast enzymes. The expression of each AtNUDT is individually regulated in different tissues. These findings suggest that most cytosolic AtNUDTs may substantially function in the sanitization of potentially hazardous ADP-ribose and the regulation of the cellular NADH/NAD(+) ratio in plant cells. On the other hand, the AtNUDT1 protein had the ability to hydrolyze 8-oxo-dGTP with a K(m) value of 6.8 mum and completely suppress the increased frequency of spontaneous mutations in the Escherichia coli mutT(-) strain, indicating that AtNUDT1 is a functional homologue of E. coli MutT in A. thaliana and is involved in the prevention of spontaneous mutation. The results obtained here suggest that the plant Nudix family has evolved in a specific manner that differs from that of yeast and humans. PMID- 15878882 TI - Remote site control of an active site fidelity checkpoint in a viral RNA dependent RNA polymerase. AB - The kinetic, thermodynamic, and structural basis for fidelity of nucleic acid polymerases remains controversial. An understanding of viral RNA-dependent RNA polymerase (RdRp) fidelity has become a topic of considerable interest as a result of recent experiments that show that a 2-fold increase in fidelity attenuates viral pathogenesis and a 2-fold decrease in fidelity reduces viral fitness. Here we show that a conformational change step preceding phosphoryl transfer is a key fidelity checkpoint for the poliovirus RdRp (3Dpol). We provide evidence that this conformational change step is orientation of the triphosphate into a conformation suitable for catalysis, suggesting a kinetic and structural model for RdRp fidelity that can be extrapolated to other classes of nucleic acid polymerases. Finally, we show that a site remote from the catalytic center can control this checkpoint, which occurs at the active site. Importantly, similar connections between a remote site and the active site exist in a wide variety of viral RdRps. The capacity for sites remote from the catalytic center to alter fidelity suggests new possibilities for targeting the viral RdRp for antiviral drug development. PMID- 15878883 TI - Chemical and biological evidence for base propenals as the major source of the endogenous M1dG adduct in cellular DNA. AB - The endogenous DNA adduct, M(1)dG, has been shown to arise in vitro in reactions of dG with malondialdehyde (MDA), a product of both lipid peroxidation and 4' oxidation of deoxyribose in DNA, and with base propenals also derived from deoxyribose 4'-oxidation. We now report the results of cellular studies consistent with base propenals, and not MDA, as the major source of M1dG under biological conditions. As a foundation for cellular studies, M1dG, base propenals, and MDA were quantified in purified DNA treated with oxidizing agents known to produce deoxyribose 4'-oxidation. The results revealed a consistent pattern; Fe2+-EDTA and gamma-radiation generated MDA but not base propenals or M1dG, whereas bleomycin and peroxynitrite (ONOO-) both produced M1dG as well as base propenals with no detectable MDA. These observations were then assessed in Escherichia coli with controlled membrane levels of polyunsaturated fatty acids (PUFA). ONOO- treatment (2 mm) of cells containing no PUFA (defined medium with 18:0/stearic acid) produced 6.5 M1dG/10(7) deoxynucleotides and no detectable lipid peroxidation products, including MDA, as compared with 3.8 M1dG/10(7) deoxynucleotides and 0.07 microg/ml lipid peroxidation products with control cells grown in a mixture of fatty acids (0.5% PUFA) mimicking Luria-Bertani medium. In cells grown with linoleic acid (18:2), the level of PUFA rose to 54% and the level of MDA rose to 0.14 microg/ml, whereas M1dG fell to 1.4/10(7) deoxynucleotides. Parallel studies with gamma-radiation revealed levels of MDA similar to those produced by ONOO- but no detectable M1dG. These results are consistent with base propenals as the major source of M1dG in this model cell system. PMID- 15878884 TI - Search of factors that intermediate cytokine-induced group IIA phospholipase A2 expression through the cytosolic phospholipase A2- and 12/15-lipoxygenase dependent pathway. AB - Inducible expression of group IIA secretory phospholipase A2 (sPLA2-IIA) by interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNFalpha) is under the control of group IVA cytosolic PLA2alpha and 12/15-lipoxygenase (12/15-LOX) in rat fibroblastic 3Y1 cells. We show here that this cytokine induction of sPLA2 IIA mRNA requires de novo protein synthesis. By means of cDNA array analysis, we found that the level of the CXC chemokine MIP-2 (macrophage inflammatory protein 2) was significantly elevated in 12/15-LOX-transfected cells compared with control cells. IL-1beta/TNFalpha-stimulated induction of endogenous MIP-2 preceded that of sPLA2-IIA, and exogenous MIP-2 induced sPLA2-IIA dose dependently. Moreover, a MIP-2-specific antisense oligonucleotide and small interfering RNA attenuated the IL-1beta/TNFalpha-induced expression of sPLA2-IIA, suggesting that MIP-2 is an absolute intermediate requirement for optimal induction of sPLA2-IIA. In addition, the expression of c-jun and fra-1, which are components of the transcription factor AP-1, was elevated in 12/15-LOX transfected cells, in which cytokine-dependent binding of AP-1 to the sPLA2-IIA promoter was increased significantly. Conversely, the receptors for transforming growth factor-beta and platelet-derived growth factor, which contributed to down regulation of sPLA2-IIA expression, were decreased following 12/15-LOX overexpression. Taken together, 12/15-LOX-dependent up-regulation of sPLA2-IIA expression may result from the interplay between accelerated MIP-2 signaling, AP 1 activation, and attenuated transforming growth factor-beta and platelet-derived growth factor signaling. PMID- 15878886 TI - Generation of normal progeny by intracytoplasmic sperm injection following grafting of testicular tissue from cloned mice that died postnatally. AB - Animal cloning by nuclear transfer has been successful in several species and was expected to become an alternative reproductive technique. Among the problems associated with this cloning technique, however, are its low success rate and high mortality of cloned animals even if they develop to term. Nuclear transfer has thus come to be considered too difficult to apply as a reproductive technique. The transplantation of male germ cells or pieces of testicular tissue has enabled the induction of spermatogenesis from fetal or postnatal male mice. In the present study, we examined whether functional male gametes could be obtained by the transplantation of pieces of testicular tissue from cloned mice that died immediately after birth with typical aberrant phenotypes, such as large offspring syndrome. Donor testicular tissues were retrieved from cloned mice that died postnatally and were transplanted into the testes of recipient nude mice. Two to three months after transplantation, the grafted donor testicular tissue had grown in the host testis, and histological analysis showed that spermatogenesis occurred within the graft. Intracytoplasmic sperm injection demonstrated that the testicular sperm generated in the grafted donor tissue were able to support full-term development of progeny. These results clearly showed that functional spermatogenesis could be induced by transplanting testicular tissue from cloned mice that died postnatally into recipient mice. The strategy presented here will be applicable to cloned animals of other species, because the xenografting of testicular tissue into mice has been demonstrated previously to be possible. PMID- 15878887 TI - Pathways participating in activation of mouse uterine natural killer cells during pregnancy. AB - Activated natural killer (NK) cells proliferate in large numbers in murine mesometrial endometrium from Day 6 to Day 12 of gestation (term = 19 gestation days) to become the most abundant uterine lymphocytes. Early human decidua contains analogous CD56+/CD16- cells. Murine uterine (u)NK cells localize to decidua basalis and mesometrial lymphoid aggregate of pregnancy (MLAp). Decidua and MLAp are transient, pregnancy-associated tissues traversed by maternal arteries to the placentas. Uterine NK cells sensitize these arteries, facilitating their structural changes into high-volume conduits by Gestation Day 10 through release of interleukin (IL)-18, interferon (IFN)-gamma, vascular endothelial growth factor (VEGF), and other molecules. Little information exists concerning where, when, or how murine or human uNK cells become activated. In murine lymphoid tissue, three NK cell adaptor-mediated activation pathways are known: FcRgamma/CD3zeta, DNAX-activating protein (DAP) 10, and DAP12 (genes Fcgr3/Cd3z, Hcst, and Tyrobp, respectively). Expression of ligands for these receptors was demonstrated in implantation sites of normal C57BL/6J mice. Then, histological and morphometric analyses of implantation sites in mice with genetic inactivation of each pathway were undertaken. Implantation sites in DAP10-/- (Hcst deleted) mice appeared normal, spiral artery modification occurred, and concentrations of IFN-gamma in MLAp and decidua basalis were similar to those in time-matched C57BL/6J. Implantation sites of FcRgamma-/-/CD3zeta-/- (Fcgr3/Cd3z double knockout), DAP12 (Tyrobp)-loss-of-function-mutant, and FcRgamma-/-/DAP12-/ (Fcgr3/Tyrobp double knockout) mice differentiated abundant but functionally impaired uNK cells that could not modify spiral arteries. These data reveal key importance of FcRgamma-/-/CD3zeta-/- and thus maternal IgG during activation of mouse uNK cells and assign DAP12 but not DAP10 signaling contributions. PMID- 15878888 TI - Calcium/calmodulin and calmodulin kinase II stimulate hyperactivation in demembranated bovine sperm. AB - Hyperactivated motility is observed among sperm in the mammalian oviduct near the time of ovulation. It is characterized by high-amplitude, asymmetrical flagellar beating and assists sperm in penetrating the cumulus oophorus and zona pellucida. Elevated intracellular Ca2+ is required for the initiation of hyperactivated motility, suggesting that calmodulin (CALM) and Ca2+/CALM-stimulated pathways are involved. A demembranated sperm model was used to investigate the role of CALM in promoting hyperactivation. Ejaculated bovine sperm were demembranated and immobilized by brief exposure to Triton X-100. Motility was restored by addition of reactivation medium containing MgATP and Ca2+, and hyperactivation was observed as free Ca2+ was increased from 50 nM to 1 microM. However, when 2.5 mM Ca2+ was added to the demembranation medium to extract flagellar CALM, motility was not reactivated unless exogenous CALM was readded. The inclusion of anti-CALM IgG in the reactivation medium reduced the proportion hyperactivated in 1 microM Ca2+ to 5%. Neither control IgG, the CALM antagonist W-7, nor a peptide directed against the CALM-binding domain of myosin light chain kinase (MYLK2) inhibited hyperactivation. However, when sperm were reactivated in the presence of CALM kinase II (CAMK2) inhibiting peptides, hyperactivation was reduced by 75%. Furthermore, an inhibitor of CAMK2, KN-93, inhibited hyperactivation without impairing normal motility of intact sperm. CALM and CAMK2 were immunolocalized to the acrosomal region and flagellum. These results indicate that hyperactivation is stimulated by a Ca2+/CALM pathway involving CAMK2. PMID- 15878889 TI - Linking two immuno-suppressive molecules: indoleamine 2,3 dioxygenase can modify HLA-G cell-surface expression. AB - Nonclassical human leukocyte antigen (HLA) class I molecule HLA-G and indoleamine 2,3 dioxygenase (INDO) in humans and mice, respectively, have been shown to play crucial immunosuppressive roles in fetal-maternal tolerance. HLA-G inhibits natural killer and T cell function by high-affinity interaction with inhibitory receptors, and INDO acts by depleting the surrounding microenvironment of the essential amino acid tryptophan, thus inhibiting T cell proliferation. We investigated whether HLA-G expression and INDO function were linked. Working with antigen-presenting cell (APC) lines and monocytes, we found that functional inhibition of INDO by 1-methyl-tryptophan induced cell surface expression of HLA G1 by HLA-G1-negative APCs that were originally cell-surface negative, and that in reverse, the functional boost of INDO by high concentrations of tryptophan induced a complete loss of HLA-G1 cell surface expression by APCs that were originally cell-surface HLA-G1-positive. This mechanism was shown to be posttranslational because HLA-G protein cell contents remained unaffected by the treatments used. Furthermore, HLA-G cell surface expression regulation by INDO seems to relate to INDO function, but not to tryptophan catabolism itself. Potential implications in fetal-maternal tolerance are discussed. PMID- 15878890 TI - The mouse epididymal transcriptome: transcriptional profiling of segmental gene expression in the epididymis. AB - Maturation of spermatozoa, including the acquisition of motility and the ability to undergo capacitation, occurs during transit through the dynamic environment of the epididymis. The microenvironments created along the length of the epididymal tubule are essential to the molecular modifications of spermatozoa that result in fertile gametes. The secretory and resorptive processes of the epithelial cells that line this tubule generate these microenvironments. In the current study, 10 morphologically distinct segments of the mouse epididymis were identified by microdissection. We hypothesized that the changing environments of the epididymal lumen are established by differential gene expression among these segments. RNA isolated from each of the 10 segments was analyzed by microarray analysis. More than 17,000 genes are expressed in the mouse epididymis, compared with about 12,000 genes identified from whole epididymal samples. Screening a panel of normal mouse tissues identified both epididymal-selective and epididymal-specific transcripts. In addition, this study identified 2168 genes that are up-regulated or down-regulated by greater than 4-fold between at least two different segments. The expression patterns of these genes identify distinct patterns of segmental regulation. Using principal component analysis, we determined that the 10 segments form 6 different transcriptional units. These analyses elucidate the changes in gene expression along the length of the epididymis for 17,000 expressed transcripts and provide a powerful resource for the research community in future studies of the biological factors that mediate epididymal sperm maturation. PMID- 15878891 TI - In utero and lactational exposure to an environmentally relevant organochlorine mixture disrupts reproductive development and function in male rats. AB - We hypothesized that in utero and lactational exposure of male rats to a mixture of more than 15 organochlorines, resembling that found in blubber from northern Quebec seals, alters reproductive development and function. Female rats were gavaged with either corn oil (controls) or the organochlorine mixture in increasing doses (low, medium, and high) for 5 wk before mating and through gestation. Developmental effects were monitored in the male offspring from Postnatal Day (PND) 2 until PND 90. The high-dose mixture reduced the number of pups per litter, percentage of live offspring, and pup weights (P < 0.05). Because only three rats from the high-dose treatment survived, data from this group beyond PND 2 were not included in the statistical analyses. As assessed by the time of preputial separation, puberty was delayed in the pups from treated dams (P < 0.05). Testes weights in the medium-dose group were greater than those in controls on PND 21 (P < 0.05). Ventral prostate weights were lower for the medium-dose group on PND 60 (P < 0.05). On PND 90, weights of the epididymis, ventral prostate, and seminal vesicle of the medium-dose rats were reduced compared to those of controls (P < 0.05). On PND 90, sperm motility parameters assessed by computer-assisted sperm analysis were altered in the low- and medium dose groups (P < 0.05). Testicular and epididymal morphology was severely affected in rats exposed to the high dose of the mixture. Serum testosterone, LH, FSH, prolactin, and total thyroxine levels did not differ because of organochlorine treatment. Therefore, in utero and lactational exposure to an environmentally relevant organochlorine mixture adversely affects the reproductive system of male rats, perhaps via antiandrogenic effects during testis development, suggesting a possible reproductive health hazard for humans and other species. PMID- 15878892 TI - Pumphead--or not! Does avoiding cardiopulmonary bypass for coronary artery bypass surgery result in less brain damage? PMID- 15878893 TI - Audit of double-lumen endobronchial intubation. PMID- 15878894 TI - Recombinant factor VIIa fails to correct coagulopathy induced by haemodilution with colloid. PMID- 15878895 TI - What does cerebral oximetry measure? PMID- 15878896 TI - Does external leg rotation facilitate femoral venipuncture? PMID- 15878897 TI - Minimum effective local anaesthetic dose for spinal anaesthesia. PMID- 15878898 TI - External iliac vein injury during total hip arthroplasty resulting in delayed shock. PMID- 15878899 TI - Insulin-sensitizing agents: use in pregnancy and as therapy in polycystic ovary syndrome. AB - Treatment with insulin-sensitizing agents is a relatively recent therapeutic strategy in women with polycystic ovary syndrome (PCOS) and insulin resistance. The key areas addressed in this review include PCOS and the development of type 2 diabetes mellitus and gestational diabetes, as well as the use of insulin sensitizing agents, particularly metformin, in the management of infertility in obese and non-obese PCOS women. Treatment with metformin in PCOS women undergoing IVF and the use of metformin during gestation will be discussed. The challenge for the health care professional should be the appropriate utilization of pharmacotherapies to improve insulin sensitivity and lower circulating insulin levels resulting in beneficial changes in PCOS phenotype. Further research into the potential role of other insulin-sensitizing agents, such as pioglitazone and rosiglitazone, in the treatment of infertile women with PCOS is needed. PMID- 15878900 TI - AMH and AMH receptor defects in persistent Mullerian duct syndrome. AB - Anti-Mullerian hormone (AMH) produced by fetal Sertoli cells is responsible for regression of Mullerian ducts, the anlage for uterus and Fallopian tubes, during male sex differentiation. A member of the transforming growth factor-beta superfamily, AMH signals through two transmembrane receptors, type II which is specific and type I receptors, shared with the bone morphogenetic protein family. Mutations of the AMH and AMH receptor type II (AMHR-II) genes lead to persistence of the uterus and Fallopian tubes in males. Both conditions are transmitted according to a recessive autosomal pattern and are symptomatic only in males. Affected individuals are otherwise normally virilized, undergo normal male puberty; and may be fertile if testes, tightly attached to the Fallopian tubes, can be replaced in the scrotum. Approximately 85% of the cases are due, in similar proportions, to mutations of the AMH or AMHR-II gene. The genetic background does not influence the phenotype, the only difference is the level of circulating AMH which is normal for age in AMHR-II mutants and usually low or undetectable in AMH gene defects. This is due to lack of secretion, explained by the localization of the mutations in critical regions, based on the assumed 3D structure of the molecule. Similarly, lack of translocation to the surface membrane is responsible for the inactivity of AMHR-II molecules bearing mutations in the extracellular domain. In 15% of cases, the cause of the persistent Mullerian duct syndrome is unknown and could be related to complex malformations of the urogenital region, unrelated to AMH physiology. PMID- 15878901 TI - Expression of interferon beta in synovial tissue from patients with rheumatoid arthritis: comparison with patients with osteoarthritis and reactive arthritis. AB - BACKGROUND: IFNbeta may have immunomodulatory effects in rheumatoid arthritis (RA) and its increased production in RA synovium may be a reactive attempt to inhibit inflammation. OBJECTIVE: To determine the expression of IFNbeta in the synovial tissue of patients with RA, osteoarthritis, and reactive arthritis. METHODS: Synovial biopsy specimens were obtained by arthroscopy from patients with RA and disease controls for immunohistological analysis using a monoclonal antibody specific for IFNbeta. Bound antibody was detected by an immunoperoxidase method. Stained sections were evaluated by computer assisted image analysis. Double stainings were performed with antibodies to detect CD55 positive fibroblast-like synoviocytes (FLS), CD68 positive macrophages, and CD83 positive dendritic cells (DCs) co-expressing IFNbeta. RESULTS: IFNbeta protein was abundantly expressed in the synovium of patients with RA. Digital image analysis showed a significant increase in the mean integrated optical density for IFNbeta expression in RA synovial tissue compared with disease controls. Specific up regulation of IFNbeta expression was also seen when the results were controlled for cell numbers. Phenotypic analysis showed that FLS, especially, but also macrophages and DCs may express IFNbeta in RA synovial tissue. CONCLUSIONS: The increased expression of IFNbeta in RA synovium suggests activation of an immunomodulatory mechanism that could inhibit synovial inflammation. PMID- 15878902 TI - EULAR report on the use of ultrasonography in painful knee osteoarthritis. Part 2: exploring decision rules for clinical utility. AB - BACKGROUND: Synovial inflammation (as defined by hypertrophy and effusion) is common in osteoarthritis (OA) and may be important in both pain and structural progression. OBJECTIVE: To determine if decision rules can be devised from clinical findings and ultrasonography (US) to allow recognition of synovial inflammation in patients with painful knee OA. METHODS: A EULAR-ESCISIT cross sectional, multicentre study enrolled subjects with painful OA knee who had clinical, radiographic, and US evaluations. A classification and regression tree (CART) analysis was performed to find combinations of predictor variables that would provide high sensitivity and specificity for clinically detecting synovitis and effusion in individual subjects. A range of definitions for the two key US variables, synovitis and effusion (using different combinations of synovial thickness, depth, and appearance), were also included in exploratory analyses. RESULTS: 600 patients with knee OA were included in the analysis. For both knee synovitis and joint effusion, the sensitivity and specificity were poor, yielding unsatisfactory likelihood ratios (75% sensitivity, 45% specificity, and positive LR of 1.36 for knee synovitis; 71.6% sensitivity, 43.2% specificity, and positive LR of 1.26 for joint effusion). The exploratory analyses did not improve the sensitivity and specificity (demonstrating positive LRs of between 1.26 and 1.57). CONCLUSION: Although it is possible to determine clinical and radiological predictors of OA inflammation in populations, CART analysis could not be used to devise useful clinical decision rules for an individual subject. Thus sensitive imaging techniques such as US remain the most useful tool for demonstrating synovial inflammation of the knee at the individual level. PMID- 15878903 TI - EULAR report on the use of ultrasonography in painful knee osteoarthritis. Part 1: prevalence of inflammation in osteoarthritis. AB - OBJECTIVES: To assess the prevalence of inflammation in subjects with chronic painful knee osteoarthritis (OA), as determined by the presence of synovitis or joint effusion at ultrasonography (US); and to evaluate the correlation between synovitis, effusion, and clinical parameters. METHODS: A cross sectional, multicentre, European study was conducted under the umbrella of EULAR-ESCISIT. SUBJECTS: had primary chronic knee OA (ACR criteria) with pain during physical activity >or=30 mm for at least 48 hours. Clinical parameters were collected by a rheumatologist and an US examination of the painful knee was performed by a radiologist or rheumatologist within 72 hours of the clinical examination. Ultrasonographic synovitis was defined as synovial thickness >or=4 mm and diffuse or nodular appearance, and a joint effusion was defined as effusion depth >or=4 mm. RESULTS: 600 patients with painful knee OA were analysed. At US 16 (2.7%) had synovitis alone, 85 (14.2%) had both synovitis and effusion, 177 (29.5%) had joint effusion alone, and 322 (53.7%) had no inflammation according to the definitions employed. Multivariate analysis showed that inflammation seen by US correlated statistically with advanced radiographic disease (Kellgren-Lawrence grade >or=3; odds ratio (OR)=2.20 and 1.91 for synovitis and joint effusion, respectively), and with clinical signs and symptoms suggestive of an inflammatory "flare", such as joint effusion on clinical examination (OR=1.97 and 2.70 for synovitis and joint effusion, respectively) or sudden aggravation of knee pain (OR=1.77 for joint effusion). CONCLUSION: US can detect synovial inflammation and effusion in painful knee OA, which correlate significantly with knee synovitis, effusion, and clinical parameters suggestive of an inflammatory "flare". PMID- 15878904 TI - Autoantibody profiling as early diagnostic and prognostic tool for rheumatoid arthritis. AB - BACKGROUND: Early treatment prevents progression of joint damage in rheumatoid arthritis (RA), but diagnosis in early disease is impeded by lack of appropriate diagnostic criteria. OBJECTIVE: To study the value of rheumatoid factor (RF), anti-cyclic citrullinated peptide autoantibodies (anti-CCP), and anti-RA33 autoantibodies for diagnosis of RA and prediction of outcome in patients with very early arthritis. METHODS: The prospective follow up inception cohort included 200 patients with very early (<3 months) inflammatory joint disease. Autoantibodies were measured at baseline and analysed in a tree based model which aimed at determining the added diagnostic value of testing for anti-CCP and anti RA33 as compared with RF alone. RESULTS: RA was diagnosed in 102 patients, while 98 developed other inflammatory arthropathies. Receiver operator curve analysis showed an optimum cut off level for RF at 50 U/ml, above which anti-CCP and anti RA33 had no additional diagnostic value. Remarkably, RF >or=50 U/ml and anti-CCP showed similar sensitivity and high specificity for RA, but overlapped considerably. Anti-RA33 was less specific and did not correlate with RF or anti CCP. Among patients with RA, 72% showed at least one of these three autoantibodies, compared with 15% of non-RA patients. RF >or=50 U/ml and anti-CCP were predictors of erosive disease, whereas anti-RA33 was associated with mild disease. CONCLUSIONS: Stepwise autoantibody testing in early inflammatory joint disease, starting with RF, followed by anti-CCP (in patients with RF <50 U/ml), and finally anti-RA33, should be used as a sensitive and effective strategy for distinguishing patients with RA at high risk for poor outcome. PMID- 15878906 TI - Enhanced gene transfer to arthritic joints using adeno-associated virus type 5: implications for intra-articular gene therapy. AB - BACKGROUND: Gene therapy of the joint has great potential as a new therapeutic approach for the treatment of rheumatoid arthritis (RA). The vector chosen is of crucial importance for clinical success. OBJECTIVE: To investigate the tropism and transduction efficiency in arthritic joints in vivo, and in synovial cells in vitro, using five different serotypes of recombinant adeno-associated virus (rAAV) encoding beta-galactosidase or green fluorescent protein genes. METHODS: rAAV was injected into the ankle joints of rats with adjuvant arthritis after the onset of disease. Synovial tissue was examined at different time points for beta galactosidase protein and gene expression by in situ staining and polymerase chain reaction (PCR) analysis, respectively. In addition, the ability of rAAV to transduce primary human fibroblast-like synoviocytes from patients with RA was investigated in vitro. RESULTS: Intra-articular injection of the rAAV5 serotype resulted in the highest synovial transduction, followed by much lower expression using rAAV2. Expression of the transgene was already detectable 7 days after injection and lasted for at least 4 weeks. Only background staining was seen for serotypes 1, 3, and 4. Importantly, there was a minimal humoral immune response to rAAV5 compared with rAAV2. Additionally, it was found that both rAAV2 and rAAV5 can efficiently transduce human fibroblast-like synoviocytes obtained from patients with RA. CONCLUSION: Intra-articular rAAV mediated gene therapy in RA might be improved by using rAAV5 rather than other serotypes. PMID- 15878905 TI - Dendritic cells, Fc{gamma} receptors, and Toll-like receptors: potential allies in the battle against rheumatoid arthritis. AB - Recent findings suggest an important role for Fcgamma receptors and Toll-like receptors expressed by dendritic cells (DC) in the pathogenesis of rheumatoid arthritis (RA). Possibly, DC behaviour might be tuned to counteract the misbalanced immune system in RA. Understanding the precise mechanisms that determine the skewed immune response in RA may provide new clues for the therapeutic use of DC in RA. PMID- 15878907 TI - Dendritic cells from patients with rheumatoid arthritis lack the interleukin 13 mediated increase of Fc gamma RII expression, which has clear functional consequences. AB - BACKGROUND: Dendritic cell (DC) function is largely tailored by Fc gamma receptors (Fc gamma R) and is critical for every immune response. OBJECTIVE: To compare interleukin (IL) 13 mediated regulation of Fc gamma RII and its related DC function between healthy controls and patients with rheumatoid arthritis (RA). METHODS: DC were derived from peripheral blood mononuclear cells according to standardised protocols. F cgammaRI, II, and III expression and DC phenotype were assessed by FACS analysis. The level of cytokine production and chemokine expression was measured by Luminex and real time quantitative polymerase chain reaction techniques. Antigen uptake capacity was studied by DC fluorescent heat aggregated immunoglobulins and FACS analysis. RESULTS: Replacement of IL4 by IL13 clearly increased the expression of Fc gamma RII on DC from healthy controls (CDC), but had no effect on DC from patients with RA (RADC). The lower production of inflammatory mediators by IL13 CDC upon Fc gamma R mediated triggering suggests that IL13 induces up regulation of specifically Fc gamma RII. RADC co cultured with IL4 already displayed an inhibitory DC phenotype, but this inhibitory phenotype was not augmented by the addition of IL13. The defective Fc gamma RII regulation was further substantiated by the finding that IL13 CDC increased antigen uptake capacity, whereas IL13 RADC did not. CONCLUSION: IL13 regulates the expression of inhibitory Fc gamma RII in normal subjects but not in RA, potentially resulting in a chronic proinflammatory immune reaction in RA. Unravelling the underlying mechanisms of Fc gamma RII regulation might lead to new therapeutic targets in RA. PMID- 15878908 TI - Mortality of rheumatoid arthritis in Japan: a longitudinal cohort study. AB - OBJECTIVE: To determine the mortality risk of Japanese patients with rheumatoid arthritis, taking into account lifestyle and physical factors, including comorbidity. METHODS: 91 individuals with rheumatoid arthritis were identified during screening a cohort of 16 119 Japanese atomic bomb survivors in the period 1958 to 1966. These individuals and the remainder of the cohort were followed for mortality until 1999. Mortality risk of the rheumatoid patients was estimated by the Cox proportional hazards model. In addition to age and sex, lifestyle and physical factors such as smoking status, alcohol consumption, blood pressure, and comorbidity were included as adjustment factors for the analysis of total mortality and for analysis of mortality from each cause of death. RESULTS: 83 of the rheumatoid patients (91.2%) and 8527 of the non-rheumatoid controls (52.9%) died during mean follow up periods of 17.8 and 28.0 years, respectively. The age and sex adjusted hazard ratio for mortality in the rheumatoid patients was 1.60 (95% confidence interval, 1.29 to 1.99), p < 0.001. Multiple adjustments, including for lifestyle and physical factors, resulted in a similar mortality hazard ratio of 1.57 (1.25 to 1.94), p < 0.001. Although mortality risk tended to be higher in male than in female rheumatoid patients, the difference was not significant. Pneumonia, tuberculosis, and liver disease were significantly increased as causes of death in rheumatoid patients. CONCLUSIONS: Rheumatoid arthritis is an independent risk factor for mortality. Infectious events are associated with increased mortality in rheumatoid arthritis. PMID- 15878909 TI - Nitrite enhances neutrophil-induced DNA strand breakage in pulmonary epithelial cells by inhibition of myeloperoxidase. AB - Chronic inhalation of environmental particles is associated with pulmonary carcinogenesis. Although the mechanism has not yet been fully elucidated, influx of inflammatory cells, including neutrophils, is suggested to play a major role in this process. Typically, in the particle-exposed lung, influx of neutrophils is accompanied by an accumulation of nitrite. Previous studies indicated that nitrite may affect the toxicity of neutrophils, involving an interaction with neutrophil-derived myeloperoxidase (MPO). To evaluate the possible consequences of this interaction for inflammation-mediated genotoxicity, we investigated the effect of nitrite on neutrophil-induced DNA damage in pulmonary target cells. Therefore, activated neutrophils were co-cultured with alveolar type II epithelial cells (RLE), and DNA strand breakage was evaluated using single-cell gel electrophoresis (comet assay). In this system, addition of nitrite caused an increase in neutrophil-induced DNA strand breakage in RLE cells, which was associated with an inhibition of MPO activity. Similar results were obtained by co-culturing RLE cells with neutrophils in the presence of the specific MPO inhibitor 4-aminobenzoic acid hydrazide (4-ABAH). To further investigate the mechanism underlying these observations, in vitro experiments were performed using mixtures of nitrite, MPO and its substrate H2O2. DNA strand breakage by reagent H2O2 was inhibited when it was allowed to react with MPO before addition to the RLE cells. However, when MPO and H2O2 were pre-mixed in the presence of nitrite or 4-ABAH, the inhibitory effect of MPO on resultant DNA damage was reversed. Further studies using catalase indicated that DNA strand breakage by the pre-mixtures of MPO, H2O2 and nitrite was H2O2-specific, suggesting that nitrite prevents consumption of H2O2 by MPO. Collectively, our results show that nitrite enhances neutrophil-induced DNA strand breakage in pulmonary epithelial cells. This effect is probably due to an inhibition of MPO activity, which increases the availability of its DNA strand breaking substrate H2O2. PMID- 15878910 TI - Polymorphisms in DNA repair genes in the molecular pathogenesis of esophageal (Barrett) adenocarcinoma. AB - To test the hypothesis that aberrations of DNA repair contribute to susceptibility for the progression of gastroesophageal reflux disease (GERD) into Barrett esophagus (BE) and esophageal adenocarcinoma (EADC), we studied the frequency of polymorphisms of selected DNA repair genes in patients with GERD (n = 126), BE (n = 125) and EADC (n = 56) enrolled in a 2-year prospective case control study. Controls comprised 95 strictly asymptomatic healthy individuals. Using genomic DNA extracted from blood samples, we identified wild-type and polymorphic variants of XPD (Arg156Arg and Lys751Gln), XRCC1 (Arg194Trp and Arg399Gln) and XRCC3 (Thr241Met), and the poly (AT) insertion/deletion of XPC (PAT). Allelic frequencies were compared between cases and controls using logistic regression to calculate age, gender, smoking and alcohol-adjusted odds ratios (OR) and 95% confidence intervals (CI). Patients with EADC demonstrated a significantly higher frequency of the XPC PAT homozygous variant genotype compared with asymptomatic controls (OR = 3.82; 95% CI = 1.05-13.93). Significantly reduced frequencies were seen for the XPD Lys751Gln homozygous variant genotype in patients with EADC (OR = 0.24; 95% CI = 0.07-0.88), and for the XRCC1 Arg399Gln homozygous variant genotype in patients with BE (OR = 0.38; 95% CI = 0.12-0.64) and GERD (OR = 0.29; 95% CI = 0.12-0.66). We conclude that the malignant phenotype probably results from a summation of polymorphic nucleotide excision repair genes showing opposing effects (an increased risk of XPC versus a protective effect of XPD). The protective effect of the homozygous variant of XRCC1 Arg399Gln for GERD and BE suggests that base excision repair alterations may occur early in progression to EADC, likely in response to GERD induced endogenous oxidative or inflammatory DNA damage. As GERD and BE are highly prevalent in the general population, this protective effect may well explain why only a fraction of individuals with GERD and BE progress into invasive EADC. PMID- 15878911 TI - Effect of inflammation on cyclooxygenase (COX)-2 expression in benign and malignant oesophageal cells. AB - Chronic inflammation has been linked to carcinogenesis in various tissue sites. Barrett's oesophageal epithelium (BE) is a premalignant condition in which cyclooxygenase-2 (COX-2) levels are increased. However, it is not clear whether the primary stimulus for the high COX-2 levels is related to inflammation or malignancy. The effect of exogenous cytokines (IL-1beta, IL-10 and IL-13) on COX 2 expression was assessed by western blotting in three BE cancer cell lines (SEG 1, BIC-1 and OE33) and a squamous cancer cell line (OE21). Primary tissue was assessed from 17 patients with long BE segments, 13 oesophagitis, 30 oesophageal adenocarcinoma (EAC), and 40 normal oesophageal (NE) and duodenal (DU) controls. COX-2 protein expression was determined by western blotting and its tissue localization was examined using immunohistochemistry. COX-2 protein and the neutrophil marker myeloperoxidase (MPO) were quantified along BE segments. The leukocyte marker CD45 was used to identify any correlation between COX-2 expression and leukocyte cell distribution in EAC. IL-1beta induced COX-2 expression in SEG-1 cells (P < 0.05), whereas IL-10 and IL-13 had no effect. COX 2 protein levels were found to be increased in distal BE > proximal BE > oesophagitis > NE (P < 0.001). COX-2 expression in EAC was heterogeneous and the overall levels were not significantly increased. The increased COX-2 expression in distal BE was not associated with inflammation (MPO expression). In addition, there was no correlation between COX-2 and CD45 in AC. COX-2 protein expression in the oesophagus appears to be independent of the degree of inflammation. PMID- 15878912 TI - Interferon regulatory factor-1 (IRF-1) exhibits tumor suppressor activities in breast cancer associated with caspase activation and induction of apoptosis. AB - We have directly assessed the ability of interferon regulatory factor-1 (IRF-1) to act as a tumor suppressor gene in human breast cancer cells and explored whether this suppressor function is mechanistically conferred by affecting cell cycle transition, apoptosis and/or caspase activation. We have used a dual approach, measuring whether overexpression of wild-type IRF-1 or a dominant negative IRF-1 (dnIRF-1) produce opposing effects on breast cancer cell proliferation in vitro or tumorigenicity in athymic nude mice. Mechanistic studies determined the effects of blocking endogenous IRF-1 expression on cell cycle transition by flow cytometry, on apoptosis by Annexin V staining, and on caspase activation by fluorescent substrate cleavage. IRF-1 mRNA (P < or = 0.001) and protein (P < or = 0.001) are highly expressed in non-tumorigenic, normal, mammary epithelial cells, with intermediate expression in tumorigenic, but non metastatic, cells and very low expression in metastatic cell lines. In MCF-7 cells transfected with a wild-type IRF-1 (MCF-7/IRF-1), IRF-1 mRNA expression inversely correlates with the rate of cell proliferation (r = -0.91; P = 0.002). Conversely, expression of dnIRF-1 in both MCF-7 (MCF-7/dnIRF-1; p53 wild-type) and T47D cells (T47D/dnIRF-1; p53 mutant) increases cell proliferation (P < or = 0.001). In athymic nude mice, the incidence of MCF-7/IRF-1 xenografts is reduced (P = 0.045), whereas MCF-7/dnIRF-1 xenografts exhibit a significantly higher tumor incidence (P < or = 0.001). Effects of IRF-1/dnIRF-1 are mediated through changes in the rates of apoptosis and not through cell cycle regulation. MCF 7/dnIRF-1 cells exhibit a 50% decrease in basal apoptosis (P = 0.007) and a significant reduction in caspase 8 activity (P = 0.03); similar effects occur in T47D/dnIRF-1 cells, where the effects on apoptosis appear to be mediated through inhibition of caspases 3/7 (P < 0.001) and caspase 8 (P = 0.03). These data establish a functional role for IRF-1 in the growth suppression of breast cancer cells and strongly implicate IRF-1 as a tumor suppressor gene in breast cancer that acts, independent of p53, to control apoptosis. PMID- 15878913 TI - Arachidonic acid, an omega-6 fatty acid, induces cytoplasmic phospholipase A2 in prostate carcinoma cells. AB - For the past 60 years, dietary intake of essential fatty acids has increased. Moreover, the omega-6 fatty acids have recently been found to play an important role in regulation of gene expression. Proliferation of human prostate cells was significantly increased 48 h after arachidonic acid (AA) addition. We have analyzed initial uptake using nile red fluorescence and we found that the albumin conjugated AA is endocytosed into the cells followed by the induction of RNA within minutes, protein and PGE2 synthesis within hours. Here we describe that AA induces expression of cytosolic phospholipase A2 (cPLA2) in a dose-dependent manner and that this upregulation is dependent upon downstream synthesis of PGE2. The upregulation of cox-2 and cPLA2 was inhibited by flurbiprofen, a cyclooxygenase (COX) inhibitor, making this a second feed-forward enzyme in the eicosanoid pathway. Cox-2 specific inhibitors are known to inhibit colon and prostate cancer growth in humans; however, recent findings show that some of these have cardiovascular complications. Since cPLA2 is upstream in the eicosanoid pathway, it may be a good alternative for a pharmaceutical target for the treatment of cancer. PMID- 15878914 TI - The effects of L-748706, a selective cyclooxygenase-2 inhibitor, on N nitrosomethylbenzylamine-induced rat esophageal tumorigenesis. AB - Epidemiological studies suggest that the frequent intake of non-steroidal anti inflammatory drugs (NSAIDs) is associated with a decreased risk of developing esophageal squamous cell carcinoma (SCC). This decrease is thought to correlate with the inhibition of cyclooxygenase (COX) activity. The production of prostaglandin E2 (PGE2), a major metabolite of COX, is increased in numerous human cancers including esophageal SCC, therefore, inhibition of COX activity and subsequent suppression of the formation of PGE2 may be chemopreventive in the esophagus. The objective of the present study was to determine whether L-748706 (L-706), a novel selective COX-2 inhibitor, would prevent N nitrosomethylbenzylamine (NMBA)-induced esophageal tumor progression in the Fischer 344 (F344) rat. In rats pretreated with a low-dose of NMBA (0.25 mg/kg body weight), L-706 at 100 p.p.m. in the diet significantly reduced tumor multiplicity but not tumor incidence. At 150 p.p.m. in the diet, L-706 alone and in combination with 200 p.p.m. piroxicam produced significant reductions in both tumor incidence and multiplicity. Inhibition of tumor development in low-dose NMBA-treated rats was associated with reductions in esophageal cell proliferation rates and PGE2 levels in preneoplastic tissues. In contrast, in rats treated with a higher dose of NMBA (0.5 mg/kg body weight), neither L-706 alone nor in combination with piroxicam reduced esophageal tumor incidence or multiplicity in spite of the fact that they reduced esophageal PGE2 levels in preneoplastic tissues and in papillomas. Cell proliferation rates were reduced only in animals treated with L-706 + piroxicam. Our data suggest that the chemopreventive treatments were effective in inhibiting tumor development in NMBA-treated animals only when they reduced PGE2 levels in preneoplastic esophageal tissues approximately to those levels found in normal esophagus. PMID- 15878915 TI - Substantial changes in gene expression of Wnt, MAPK and TNFalpha pathways induced by TGF-beta1 in cervical cancer cell lines. AB - Transforming growth factor-beta 1 (TGF-beta1) is a potent inhibitor of epithelial cell proliferation. During the development of cervical carcinoma however, an increase in production of TGF-beta1 is accompanied by decreased sensitivity for the growth-limiting effect of TGF-beta1. TGF-beta1 has an anti-proliferative effect on cells of the immune system and thus can be advantageous for tumor progression. The aim of the present study was to determine the effect of TGF beta1 on mRNA expression profile of genes in pathways involved in cell growth and cell death, in cervical carcinoma cell lines with different sensitivity to TGF beta1. For this purpose, we have investigated changes in gene expression in TGF beta1 stimulated cervical cancer cell lines with high (CC10B), intermediate (SiHa) and low (HeLa) sensitivity to the anti-proliferative effect of TGF-beta1, at timepoints 0, 6, 12 and 24 h. Microarray analysis, using Affymetrics focus arrays, representing 8973 genes, was used to measure gene expression. In our study novel target genes involved in tumor necrosis factor alpha (TNFalpha), mitogen-activated protein kinase (MAPK) and wingless type (Wnt) pathways in response to TGF-beta1 were found. Substantial differences in gene expression between TGF-beta1 sensitive and insensitive cell lines were observed involving genes in TNFalpha, MAPK, Wnt and Smad pathways. Since these pathways are implicated in cell proliferation and cell death, these pathways may play a role in determining the overall sensitivity of a cell to TGF-beta1 induced cell growth inhibition. The results were subsequently validated by quantitative real-time PCR. Increased resistance to TGF-beta1 induced cell growth inhibition was correlated with an elevated production of TGF-beta1 by the cell lines, as measured by enzyme linked immunosorbent assay. TGF-beta1 production did not inhibit cell growth, since blocking TGF-beta1 protein by anti-TGF-beta had no effect on cell proliferation. TGF-beta1 excretion by tumor cells more likely contributes to paracrine stimulation of tumor development. PMID- 15878916 TI - Genetic polymorphisms of p21 are associated with risk of squamous cell carcinoma of the head and neck. AB - The p21 (Waf1/Cip1/CDKN1A) protein regulates the transition from the G1 to the S phase and has an important role in modulating cell-cycle control, apoptosis and cell growth. Two polymorphisms of the p21 gene at codon 31 (p21 C98A, dbSNP rs1801270) and at the 3' untranslated region (p21 T70C, dbSNP rs1059234) may have an effect on the protein function and may thus play a role in the development of cancer. We hypothesized that these two p21 polymorphisms are associated with the risk of squamous cell carcinoma of the head and neck (SCCHN). We tested this hypothesis in a hospital-based case-control study of 712 patients newly diagnosed with SCCHN and 1222 cancer-free controls who were frequency-matched by age, sex and ethnicity. All subjects were non-Hispanic whites. Our results showed that the variant alleles and genotypes were more common among cases than among controls (P < 0.001 and P = 0.013 for p21C70T, and P < 0.001 and P = 0.035 for p21C98A, respectively). Compared with the p21 70CC genotype, there was a significantly greater risk of SCCHN associated with the variant p21 70TC [odds ratio (OR) = 1.47, 95% confidence interval (CI) = 1.12-1.93] and combined p21 70TC/TT (OR = 1.49, 95% CI = 1.14-1.95) genotypes. Similarly, compared with the p21 98CC genotype, there was also a significantly greater SCCHN risk associated with the variant p21 98AC (OR = 1.32, 95% CI = 1.00-1.73) and combined p21 98AC/AA (OR = 1.37, 95% CI = 1.05-1.79) genotypes. When these two polymorphisms were evaluated together by the number of risk alleles, there was a significant increase in SCCHN risk that was dependent on the number of risk alleles (P(trend) = 0.001). Our results suggest that the presence of these two p21 polymorphisms may be a marker of genetic susceptibility to SCCHN. PMID- 15878917 TI - Comparison of male and female meiotic segregation patterns in translocation heterozygotes: a case study in an animal model (Sus scrofa domestica L.). AB - BACKGROUND: The comparison of male and female meiotic segregation patterns for individuals carrying identical reciprocal translocations has been rarely reported in mammalian species. The main comparative study involving males and females with comparable genetic background has been performed in the mouse. Swine is another relevant animal model species for meiotic studies. Here we present the segregation patterns determined for sows carrying one of the two following reciprocal translocations: 38, XX, rcp(3;15)(q27;q13), and 38, XX, rcp(12;14)(q13;q21). These segregation data were compared to those previously obtained for closely related boars carrying the same balanced chromosomal rearrangements. METHODS: Dual colour in situ hybridization of whole chromosome painting probes was carried out on metaphases of in vitro-matured oocytes II. Segregation results were obtained for 118 and 206 metaphases II respectively for the two translocations. RESULTS: Significant differences between sexes were demonstrated for both rearrangements. For instance, for the 3/15 translocation, the chromosomally unbalanced gametes were of different origin: preponderance of the adjacent-I segregation in the male (31.4%), and of the adjacent-II (14.3%) and 3:1 (14.3%) segregations in females. For the 12/14 translocation, the proportion of balanced gametes was greater in males than in females (75.9 and 59.4% respectively). CONCLUSION: This study is a new scientific contribution to compare the segregation patterns of male and female carriers of identical chromosomal rearrangements. The results obtained are consistent with those previously reported in mice. Hypotheses to interpret the observed differences between the two translocations, as well as between the male and female segregation patterns, are formulated and discussed. PMID- 15878918 TI - Increased expression of Leydig cell haem oxygenase-1 preserves spermatogenesis in varicocele. AB - BACKGROUND: Oxidative stress is involved in the pathogenesis of testicular disorders. Haem oxygenase-1 (HO-1) plays an important cytoprotective role against oxidative stress. We investigated the presence of oxidative stress, represented by generation of 4-hydroxy-2-nonenal (4-HNE)-modified proteins, and expression of HO-1 in varicocele testes of human. METHODS: Thirty testicular biopsies from patients with left varicocele and 10 from patients with normal spermatogenesis were included. Generation of 4-HNE-modified proteins was examined as a marker of oxidative stress. Expression of HO-1 was assessed by western blotting and immunohistochemistry. The expression was compared with clinico-pathological parameters. RESULTS: Increased generation of 4-HNE-modified proteins was observed in varicocele testes. HO-1 expression was significantly correlated with varicocele grade (P < 0.01 in grade 2 and 3 compared to control) and expression of 4-HNE-modified proteins (r = 0.508, P < 0.01). The immunoreactivity was increased in Leydig cells in varicocele testes. There were significant correlations between age, total motile sperm count, Johnsen's mean score and HO 1:4-HNE-modified protein ratio (r = 0.206, 0.405 and 0.408, P = 0.027, 0.027 and 0.025 respectively). CONCLUSIONS: In testes with varicocele, there are increases in 4-HNE-modified proteins, suggesting that oxidative stress is present. Increased HO-1 expression, mainly in Leydig cells, is considered to protect the cells against oxidative stresses in varicocele testes. PMID- 15878919 TI - CYP17 genotype predicts serum hormone levels among pre-menopausal women. AB - BACKGROUND: CYP17, which encodes cytochrome P450c17alpha, mediates both steroid 17alpha-hydroxylase and 17,20-lyase activities, and is essential for the production of glucocorticoids and sex steroids. There is evidence that a common polymorphism in CYP17 (T27C) is associated with estrogen levels, making it a potential marker of disease risk. METHODS: This is the first study to examine the relationship between CYP17 and estradiol (E2) using serum sampled exclusively from the early follicular phase of the menstrual cycle. We assessed the relationship between CYP17 and serum hormone levels, menstrual cycle length, bleed length, and age at menarche in 164 pre-menopausal women. RESULTS: Among women with body mass index (BMI) < or =25 kg/m2, those with the TC and CC genotypes had 19 and 42% higher E2 (P for trend 0.007) and 14 and 30% higher dehydroepiandrosterone sulphate respectively (P for trend 0.10) than women with the TT genotype. Androstenedione levels did not differ between genotypes. Among women with BMI >25 kg/m2, hormone levels did not differ by genotype. Women with the C allele were also more likely to have menstrual cycle lengths <27 days [odds ratio (OR) for TC=2.36, 95% confidence interval (CI)=1.24-4.52; OR for CC=5.59, 95% CI=1.53-20.43 compared to TT]. CYP17 genotype was not associated with menstrual bleed length or age at menarche. CONCLUSION: The CYP17 T27C polymorphism may be a marker of endocrine function. PMID- 15878920 TI - Glutathione-S-transferase P1 gene polymorphism and susceptibility to endometriosis. AB - BACKGROUND: Glutathione-S-tranferase (GST) is the part of the key phase II detoxifying enzyme system. Many studies have investigated the role of GSTM1 and GSTT1 gene polymorphisms in endometriosis. Although GSTP1 was found to be one of the most abundant types of GST in genital system, there are insufficient data about the importance of the role of GSTP1 gene polymorphism in endometriosis. METHODS: This case-control study involved 150 patients with endometriosis and 150 controls. The frequency of GSTP1 single nucleotide polymorphisms was evaluated using PCR and melting curve analysis. RESULTS: The proportion of GSTP1 ile/ile tended to be higher in patients with endometriosis than control group, although the difference was not significant [odds ratio (OR)=1.53; 95% confidence interval (CI)=0.95-2.46]. In contrast, GSTP1 val/val was significantly higher in control patients and seems protective for endometriosis (OR=0.10; 95% CI=0.02-0.42). CONCLUSION: The results of this study suggest that GSTP1 polymorphism might modulate the risk of endometriosis with significantly decreased risk for GSTP1 val/val and marginally increased risk for GSTP1 ile/ile. Further studies on not only the disease processes but also normal distribution of the enzyme in female genital tract may provide better understanding about the role of GST types and their polymorphs in endometriosis. PMID- 15878921 TI - In search of candidate genes critically expressed in the human endometrium during the window of implantation. AB - BACKGROUND: In this prospective randomized blinded clinical trial, we examined gene expression profiles of the human endometrium during the early and mid-luteal phases of the natural cycle. METHODS: An endometrial biopsy was performed on day 16 (LH +3) or on day 21 (LH +8), followed by RNA extraction and microarray analysis using an Affymetrix HG-U95A microchip. Data analysis was carried out using pairwise multiple group comparison with the significance analysis of microarrays (SAM) software. RESULTS: With a false discovery rate of 0, the analysis revealed that 107 genes were significantly and differently expressed (> or =2-fold) during the early versus the mid-luteal phase of the cycle. Forty-five of these genes have not been previously linked to endometrial receptivity. Validation of the microarray data was accomplished using semiquantitative RT-PCR. We demonstrated the presence of estrogen and progesterone response elements (ERE and PRE) by analysis of the 5'-flanking regions of a subset of differentially regulated genes. CONCLUSIONS: Using a strict bioinformatics approach of microarray data, we demonstrated significant changes in candidate genes during the transition of the early to the mid-luteal phase of the human endometrium that may have functional significance for the opening and maintenance of the window of implantation. PMID- 15878922 TI - Ultrastructural studies of spermatozoa from infertile males with Robertsonian translocations and 18, X, Y aneuploidies. AB - BACKGROUND: In order to clarify the relationship between chromosomal rearrangements, sperm morphology and interchromosomal effects (ICE), we studied the spermatogenetic defects in seven infertile Robertsonian translocation carriers. METHODS: Lymphocyte karyotypes were evaluated using Giemsa-Trypsin Giemsa banding and fluorescence in-situ hybridization (FISH). Semen analysis was performed by light and transmission electron microscopy. FISH of sperm nuclei was carried out to detect possible ICE. RESULTS: Lymphocyte karyotype analysis revealed five t(13;14), one t(13;21) and one t(14;22) carriers. Sperm ultrastructural examination highlighted a higher percentage of immaturity, apoptosis and necrosis than in controls. Aneuploidies of gonosomes were detected in sperm from five out of six carriers of Robertsonian translocation, whereas aneuploidy of chromosome 18 was evident in three out of six carriers. The frequencies of diploidy were altered in all cases. CONCLUSIONS: Since these infertile patients showed severe spermatogenetic impairment from the morphological and meiotic points of view, we recommend detailed sperm ultrastructural and chromosomal analysis before undertaking ICSI cycles in Robertsonian translocation carriers. PMID- 15878923 TI - How complete is full thickness disc resection of bowel endometriotic lesions? A prospective surgical and histological study. AB - BACKGROUND: This study aims to evaluate the completeness of full thickness disc resection in the treatment of deep endometriotic bowel lesions. METHODS: This study comprised 16 women with bowel endometriotic lesions requiring segmental resection. For the purpose of the study, before intestinal resection, nodulectomy was performed. The presence of endometriotic infiltration in direct continuity with the removed nodule and the presence of fibrosis in the area surrounding the nodule were histologically evaluated. RESULTS: In seven out of 16 cases (43.8%; 95% CI, 19.8-70.1), endometriosis was found in the bowel wall adjacent to the site of nodulectomy; the infiltration was visible in the muscular layer in all cases. In cases of incomplete nodulectomy, the muscular layer of the bowel segment surrounding the endometriotic nodule contained limited or no fibrosis. CONCLUSIONS: Full thickness disc resection is not complete in > or =40% of women with bowel endometriosis. Our finding that fibrosis in the muscular layer, the main landmark during surgical resection, does not always surround bowel endometriotic lesions might explain why incomplete resection may occur. PMID- 15878924 TI - Correlation between in vitro maturation and expression of LH receptor in cumulus cells of the oocytes collected from PCOS patients in HCG-primed IVM cycles. AB - BACKGROUND: The aim of this study was to investigate whether in vitro maturation (IVM) and blastocyst development of oocytes collected following HCG-primed IVM cycles of PCOS patients are correlated with their cumulus cell (CC) patterns and further to investigate mRNA expression of the receptors for FSH, LH and epidermal growth factor (EGF) in the CCs with each pattern. METHODS: Patients who underwent IVM were primed with 10,000 IU of HCG 36 h before oocyte aspiration. The isolated cumulus-oocyte complexes were divided into three groups according to the CC patterns: oocytes with dispersed CCs (group A), oocytes with compacted CCs (group B) and oocytes with sparse CCs (group C). Oocyte maturation and blastocyst development were compared among three groups. The expression of the mRNA for FSH, LH and EGF receptors in group A and B was analysed by semi-quantitative RT-PCR. RESULTS: The maturation rate of group A was significantly higher than those of group B and C. The rate of blastocysts in group A was significantly higher than those of group B and C. mRNA expression of the LH receptor in group A was more abundant than that of group B. CONCLUSIONS: These results suggest that the presence of dispersed CCs at oocyte collection may be positively correlated with the rates of oocyte maturation and blastocysts in HCG-primed IVM cycles. In addition, the expression of LH receptor in CCs may be correlated with the CC pattern of oocytes at collection. PMID- 15878925 TI - Clomiphene citrate--end of an era? A mini-review. AB - The purpose of this review is to examine whether the time has come to replace clomiphene citrate (CC) as the first line therapy for WHO group II (eu oestrogenic) infertility, the majority of which is associated with polycystic ovary syndrome. CC has been the first line therapy for these cases for the last 40 years. It is a simple, cheap treatment, almost devoid of side effects which yields a single live birth rate of approximately 25% of starters. Non-response to CC and the gap between ovulation and pregnancy rates have variously been attributed to its anti-estrogen effects, and high LH and androgen concentrations. Three possible contenders for the replacement of CC as first-line treatment are scrutinized: metformin, aromatase inhibitors and low-dose FSH. Each has their advantages and disadvantages, but none of them, while showing much potential promise, has been proven, as yet, to be a feasible replacement for CC in this role. For CC, it may not yet be the end of an era but it may be the beginning of the end. PMID- 15878926 TI - Vasculogenesis in complete and partial hydatidiform mole pregnancies studied with CD34 immunohistochemistry. AB - BACKGROUND: Defective chorionic villous vascularization is present in pregnancies complicated by absent or abnormal embryonic development. The aim of this study was to investigate the embryonic and/or maternal genomic influence on vasculogenesis in diploid complete hydatidiform mole (CHM) and in triploid partial hydatidiform mole (PHM) in comparison with normal development. METHODS: Mean villous stromal area and functional vascular area, vessels with a lumen and haemangiogenetic cords, peripherally or centrally located were measured and counted in chorionic villi of 12 CHM, 12 normal pregnancies (termination of pregnancy, TOP) and 15 PHM of which nine were without an embryo (PHM-E) and six were with an embryo (PHM + E), using quantitative CD34 immunohistochemistry. RESULTS: TOP showed significantly more vessels per chorionic villus, centrally and peripherally located (median, range), than CHM, PHM-E and PHM + E (4.0, 0-9 versus 0.0, 0-11, 0.0, 0-18 and 1.0, 0-21). CHM showed significantly more centrally located cords than PHM-E, PHM + E and TOP (1.5, 0-22 versus 1.0, 0-15, 0.5, 0-8 and 1.0, 0-2). CONCLUSIONS: Initiation of chorionic villous vasculogenesis is independent of the maternal genome (CHM). The development of an embryo, however, is obligatory for the modulation of normal vascularization resulting in a well developed vasculosyncytial membrane. PMID- 15878927 TI - A randomized trial of mifepristone in combination with misoprostol administered sublingually or vaginally for medical abortion at 13-20 weeks gestation. AB - BACKGROUND: Several studies have now reported the successful use of the sublingual administration of misoprostol for medical abortion in the first trimester. The objective of this study was to assess the acceptability to women, the efficacy of the regimen, as well as the acceptability to staff of sublingual versus vaginal administration of misoprostol following mifepristone for medical abortion at 13-20 weeks gestation. METHODS: Women were randomized by opening consecutive sealed envelopes generated using random number tables. Mifepristone (200 mg) was followed 36-48 h later by sublingual administration of misoprostol 600 microg or vaginal misoprostol 800 microg. This was followed by 3 hourly doses of misoprostol 400 microg administered sublingually or vaginally. RESULTS: A total of 76 women were randomized. Of women in the sublingual group, 24 (66.7%) expressed satisfaction with the route of misoprostol administration compared with 25 (62.5%) in the vaginal group. A higher proportion in the sublingual group used intramuscular opiates. There was no significant difference in the surgical evacuation rate between the sublingual (three out of 36 women, 8.3%) and vaginal groups (one out of 40, 2.5%), (P=0.26) and acceptability to staff was the same for both methods. CONCLUSIONS: Sublingual administration of misoprostol following mifepristone is an acceptable and effective alternative to vaginal administration for medical abortion at 13-20 weeks gestation. However, women should be advised about the greater likelihood of requiring stronger analgesia. PMID- 15878928 TI - Patients with abnormal sperm parameters have an increased sex chromosome aneuploidy rate in peripheral leukocytes. AB - BACKGROUND: Patients with oligoasthenoteratozoospermia (OAT) and normal karyotypes have an increased sperm aneuploidy rate. This may be due to an altered intratesticular environment that affects the chromosomal segregation mechanism(s). Alternatively, it may be due to a generalized meiotic and mitotic abnormality. In this case, patients with abnormal spermatogenesis should also have an increased somatic cell aneuploidy rate. To test this hypothesis, we evaluated peripheral leukocyte aneuploidy rate in patients with spermatogenic impairment. METHODS: In all, 38 patients were enrolled, of whom 20 had OAT, 15 non-obstructive azoospermia and three Y chromosome (Yq) microdeletions (AZF). Eight healthy normozoospermic men with proven fertility were recruited as controls. Conventional karyotype analysis, AZF microdeletion evaluation and triple-colour FISH for chromosomes X, Y and 12 were conducted in all patients and controls. A total of 1000 lymphocytes were scored for each patient and control. RESULTS: All patients and controls had a normal karyotype. Sex chromosome aneuploidy rates in peripheral lymphocytes was significantly higher in patients with OAT (0.74+/-0.09%), azoospermia (1.15+/-0.15%) or Yq microdeleted (1.54+/ 0.40%), compared with controls (0.15+/-0.03%) (P <0.05). CONCLUSIONS: Patients with OAT, azoospermia or Yq microdeletions had a slight, but significant, increase of sex chromosome aneuploidy rate in lymphocytes, suggesting the presence of a generalized defective cell division mechanism. In contrast with recent observations, Yq microdeletions do not seem to predispose to a higher number of malsegregation events in somatic cells compared with patients with azoospermia. PMID- 15878929 TI - Birth of a healthy infant following trophectoderm biopsy from blastocysts for PGD of beta-thalassaemia major. AB - PGD is a well accepted reproductive choice for couples at genetic risk and involves the diagnosis and transfer of unaffected IVF embryos. PGD for monogenetic diseases is most commonly accomplished by the biopsy of one or two blastomeres from cleavage stage embryos, followed by PCR-based protocols. However, PCR-based DNA analysis of one or two cells is subject to several problems, including total PCR failure, or failure of one allele to amplify. Trophectoderm biopsy at the blastocyst stage enables the removal of more than two cells for diagnosis while being non-invasive to the inner cell mass which is destined for fetal development. The aim of this study was to develop a safe, reliable technique for the biopsy of trophectoderm cells from human blastocysts. This case report demonstrates that removal of trophectoderm cells prior to blastocyst transfer is compatible with implantation and development to term. Here we report successful PGD for beta-thalassaemia following trophectoderm cell biopsy from blastocysts and the birth of a healthy infant. PMID- 15878930 TI - Levothyroxine treatment in thyroid peroxidase antibody-positive women undergoing assisted reproduction technologies: a prospective study. AB - BACKGROUND: Infertile women positive for thyroid antibodies suffer from a poor pregnancy/delivery outcome, although conflicting data have been published. Our objective was to investigate if levothyroxine (LT4) exerts any effect on pregnancy and/or delivery rates in thyroid peroxidase antibody (TPOAb)-positive (+) women undergoing assisted reproductive technologies. METHODS: Patients undergoing treatment were screened for TPOAb, thyroid-stimulating hormone (TSH) and free thyroxine (FT4). A total of 72 (15%) out of the 484 euthyroid women selected were TPOAb (+). These 72 patients were randomly divided into two groups: group A (n = 36) underwent LT4 treatment, group B (n = 36) placebo. Group C consisted of 412 women (85%) who were TPOAb negative (-). All patients received controlled ovarian stimulation. The endpoints of treatment were pregnancy rate, miscarriage rate and delivery rate. RESULTS: No differences in pregnancy rate were observed between the three groups. Miscarriage rate was higher in TPOAb (+) in comparison to TPOAb (-) [relative risk: 2.01 (95% CI = 1.13-3.56), P = 0.028]. CONCLUSIONS: The pregnancy rate is not affected either by presence of TPOAb or treatment with LT4. However, TPOAb (+) women show a poorer delivery rate compared to TPOAb (-). LT4 treatment in TPOAb (+) does not affect the delivery rate. PMID- 15878931 TI - Peroxynitrite provides the peroxide tone for PGHS-2-dependent prostacyclin synthesis in vascular smooth muscle cells. AB - Endotoxin-treated vascular smooth muscle cells (VSMCs) were recently shown to release high amounts of prostacyclin (PGI2) dependent on the induction of prostaglandin endoperoxide synthase-2 (PGHS-2). In contrast to endothelial PGI2 synthase, for which nitration and inhibition by peroxynitrite was reported, addition of SIN-1 as a peroxynitrite-generating system did not cause inhibition but rather doubled PGI2 release by VSMC. The hypothesis of peroxynitrite supplementing an unsaturated peroxide tone for PGHS-2 was supported by H2O2 exerting the same effect. Studies performed with purified PGHS-2 revealed maximal elevation of enzyme activity in the presence of equimolar concentrations of *NO and *O2-, which together form peroxynitrite, while excessive production of either one radical was inhibitory. Most importantly, 6-keto-PGF1alpha formation by intact VSMC depended on a nearly equimolar generation of *NO and *O2- for providing the endogenous peroxide tone. These findings, together with the observation that an excess of exogenously added *NO, as well as uric acid as a scavenger of peroxynitrite potently reduced PGI2 release, underlined the role of peroxynitrite as the dominating and rate-limiting intracellular mediator of peroxide tone in VSMC. The results allow us to postulate a new cross-talk between the *NO and the prostanoid pathways with a crucial role for peroxynitrite in providing the peroxide tone for a continuous activation of PGHS-2. PMID- 15878932 TI - Effects of alpha-AMPK knockout on exercise-induced gene activation in mouse skeletal muscle. AB - We tested the hypothesis that 5'AMP-activated protein kinase (AMPK) plays an important role in regulating the acute, exercise-induced activation of metabolic genes in skeletal muscle, which were dissected from whole-body alpha2- and alpha1 AMPK knockout (KO) and wild-type (WT) mice at rest, after treadmill running (90 min), and in recovery. Running increased alpha1-AMPK kinase activity, phosphorylation (P) of AMPK, and acetyl-CoA carboxylase (ACC)beta in alpha2-WT and alpha2-KO muscles and increased alpha2-AMPK kinase activity in alpha2-WT. In alpha2-KO muscles, AMPK-P and ACCbeta-P were markedly lower compared with alpha2 WT. However, in alpha1-WT and alpha1-KO muscles, AMPK-P and ACCbeta-P levels were identical at rest and increased similarly during exercise in the two genotypes. The alpha2-KO decreased peroxisome-proliferator-activated receptor gamma coactivator (PGC)-1alpha, uncoupling protein-3 (UCP3), and hexokinase II (HKII) transcription at rest but did not affect exercise-induced transcription. Exercise increased the mRNA content of PGC-1alpha, Forkhead box class O (FOXO)1, HKII, and pyruvate dehydrogenase kinase 4 (PDK4) similarly in alpha2-WT and alpha2-KO mice, whereas glucose transporter GLUT 4, carnitine palmitoyltransferase 1 (CPTI), lipoprotein lipase, and UCP3 mRNA were unchanged by exercise in both genotypes. CPTI mRNA was lower in alpha2-KO muscles than in alpha2-WT muscles at all time points. In alpha1-WT and alpha1-KO muscles, running increased the mRNA content of PGC-1alpha and FOXO1 similarly. The alpha2-KO was associated with lower muscle adenosine 5'-triphosphate content, and the inosine monophosphate content increased substantially at the end of exercise only in alpha2-KO muscles. In addition, subcutaneous injection of 5-aminoimidazole-4-carboxamide-1-beta-4 ribofuranoside (AICAR) increased the mRNA content of PGC-1alpha, HKII, FOXO1, PDK4, and UCP3, and alpha2-KO abolished the AICAR-induced increases in PGC-1alpha and HKII mRNA. In conclusion, KO of the alpha2- but not the alpha1-AMPK isoform markedly diminished AMPK activation during running. Nevertheless, exercise induced activation of the investigated genes in mouse skeletal muscle was not impaired in alpha1- or alpha2-AMPK KO muscles. Although it cannot be ruled out that activation of the remaining alpha-isoform is sufficient to increase gene activation during exercise, the present data do not support an essential role of AMPK in regulating exercise-induced gene activation in skeletal muscle. PMID- 15878933 TI - Antenatal steroids and fluid balance in very low birthweight infants. AB - OBJECTIVES: To determine if insensible water loss (IWL) differed between infants exposed or not exposed antenatally to corticosteroids and to explore possible mechanisms for the early postnatal diuresis associated with antenatal steroid exposure. DESIGN: Retrospective analysis of prospectively collected data. SETTING: Level three neonatal intensive care unit. PATIENTS: Ninety six infants, median gestational age 27.5 weeks (range 23-33). MAIN OUTCOME MEASURES: Comparison of the IWL, urine output and osmolality, fluid input, electrolyte imbalance, respiratory illness severity (as assessed by surfactant requirement, maximum peak inspiratory pressure, and inspired oxygen concentration), and cardiovascular status (as assessed by inotrope requirement) between infants with antenatal corticosteroid exposure and gestational age matched controls. RESULTS: The infants exposed to antenatal steroids differed significantly from the controls in having both a lower IWL (p = 0.0135) and a higher urine output (p = 0.0036) on day 1, and fewer developed hyponatraemia (p = 0.027) on day 2. Fewer of those exposed to antenatal steroids required inotropes (p = 0.06), but their respiratory status was similar to that of the controls. CONCLUSIONS: Infants exposed to antenatal corticosteroids have a lower IWL. The results suggest that greater skin maturation, but also better perfusion rather than less severe respiratory status, explains the early diuresis in infants exposed to antenatal steroids. PMID- 15878934 TI - Early onset neonatal meningitis in Australia and New Zealand, 1992-2002. AB - OBJECTIVES: To study the epidemiology of early onset neonatal bacterial meningitis (EONBM) in Australasia. DESIGN: Prospective surveillance study, 1992 2002, in 20 neonatal units in Australia and New Zealand. EONBM was defined as meningitis occurring within 48 hours of delivery. RESULTS: There were 852 babies with early onset sepsis, of whom 78 (9.2%) had EONBM. The incidence of early onset group B streptococcal meningitis fell significantly from a peak of 0.24/1000 live births in 1993 to 0.03/1000 in 2002 (p trend = 0.002). There was no significant change over time in the incidence of Escherichia coli meningitis. The rate of EONBM in very low birthweight babies was 1.09/1000 compared with the rate in all infants of 0.11/1000. The overall rate of EONBM was 0.41/1000 in 1992 and 0.06 in 2001, but this trend was not significant (p trend = 0.07). Case fatality rates for EONBM did not change significantly with time. Birth weight <1500 g (odds ratio (OR) 7.2 (95% confidence interval (CI) 4.8 to 10.9)) and Gram negative bacillary meningitis (OR 3.3 (95% CI 2.2 to 4.9)) were significant risk factors for mortality. Sixty two percent of the 129 babies who died from early onset sepsis or suspected sepsis did not have a lumbar puncture performed. CONCLUSION: The incidence of early onset group B streptococcal meningitis has fallen, probably because of maternal intrapartum antibiotic prophylaxis, without a corresponding change in E. coli meningitis. Gram negative bacillary meningitis still carries a worse prognosis than meningitis with a Gram positive organism. PMID- 15878935 TI - Risk factors for respiratory morbidity in infancy after very premature birth. AB - OBJECTIVES: To determine the occurrence of respiratory morbidity during infancy after very premature birth and to identify risk factors. DESIGN: Prospective follow up study. SETTING: The United Kingdom oscillation study. PATIENTS: 492 infants, all born before 29 weeks gestation. INTERVENTIONS: Structured questionnaires were completed by local paediatricians when the infants were seen in outpatients at 6 and 12 months of age corrected for prematurity. MAIN OUTCOME MEASURES: Cough, wheeze, and treatment requirements and the composite measure of respiratory morbidity (cough, frequent cough, cough without infection, wheeze, frequent wheeze, wheeze without infection, and use of chest medicine) and their relation to 13 possible explanatory variables. RESULTS: At 6 and 12 months of corrected age, 27% of the infants coughed and 6% had frequent (more than once a week) cough, and 20% and 3% respectively had wheeze or frequent wheeze. At 6 and 12 months, 14% of infants had taken bronchodilators and 8% inhaled steroids. After adjustment for multiple outcome testing, four factors were associated with increased respiratory morbidity: male sex, oxygen dependency at 36 weeks postmenstrual age, having older siblings aged less than 5 years, and living in rented accommodation. CONCLUSIONS: Male infants are particularly vulnerable to respiratory morbidity in infancy after very premature birth. It is important to identify a safe and effective strategy to prevent chronic oxygen dependency. PMID- 15878936 TI - Dampened ventilatory response to added dead space in newborns of smoking mothers. AB - BACKGROUND: Term newborns can compensate fully for an imposed dead space (tube breathing) by increasing their minute ventilation. OBJECTIVE: To test the hypothesis that infants of smoking mothers would have an impaired response to tube breathing. DESIGN: Prospective study. SETTING: Perinatal service. PATIENTS: Fourteen infants of smoking and 24 infants of non-smoking mothers (median postnatal age 37 (11-85) hours and 26 (10-120) hours respectively) were studied. INTERVENTIONS: Breath by breath minute volume was measured at baseline and when a dead space of 4.4 ml/kg was incorporated into the breathing circuit. MAIN OUTCOME MEASURES: The maximum minute ventilation during tube breathing was determined and the time constant of the response calculated. RESULTS: The time constant of the infants of smoking mothers was longer than that of the infants of non-smoking mothers (median (range) 37.3 (22.2-70.2) v 26.2 (13.8-51.0) seconds, p = 0.016). Regression analysis showed that maternal smoking status was related to the time constant independently of birth weight, gestational or postnatal age, or sex (p = 0.018). CONCLUSIONS: Intrauterine exposure to smoking is associated with a dampened response to tube breathing. PMID- 15878937 TI - 'You ain't going to say...I've got a problem down there': workplace-based prostate health promotion with men. AB - Prostate health has emerged as a key health issue for men. Nearly 10,000 men die from prostate cancer each year and many more live with non-cancerous, but debilitating, prostate conditions. Despite the widespread prevalence, evidence suggests that men lack knowledge about male cancers and conditions, and are more likely to ignore signs and delay seeking help. Health promotion with men in the workplace is, therefore, increasingly being advocated as an important way of providing men with health information and encouraging them to see a health professional where appropriate. However, there has not been a developed account of men's views on health promotion within the workplace. This paper presents the findings of a small-scale qualitative study that explored men's perceptions and experiences of three different workplace-based health promotion interventions to improve prostate health awareness and their attitudes towards the workplace as an appropriate setting for promoting men's health. This paper shows that men generally welcomed a workplace-based health promotion campaign targeted specifically at them. However, the masculine 'culture' of the workplace, where concerns about health were likely to be met with ridicule rather than concern, was one important factor in understanding these men's views of different health promotion interventions. PMID- 15878938 TI - Synaptic input of rat spinal lamina I projection and unidentified neurones in vitro. AB - Spinal lamina I projection neurones that transmit nociceptive information to the brain play a pivotal role in hyperalgesia in various animal models of inflammatory and neuropathic pain. Consistently, activity-dependent long-term potentiation can be induced at synapses between primary afferent C-fibres and lamina I projection neurones but not unidentified neurones in lamina I. The specific properties that enable projection neurones to undergo long-term potentiation and mediate hyperalgesia are not fully understood. Here, we have tested whether lamina I projection neurones differ from unidentified neurones in types or strength of primary afferent input and/or action potential-independent excitatory and inhibitory input. We used the whole-cell patch-clamp technique to record synaptic currents in projection and unidentified lamina I neurones in a transverse lumbar spinal cord slice preparation from rats between postnatal day 18 and 37. Lamina I neurones with a projection to the parabrachial area or the periaqueductal grey were identified by retrograde labelling with a fluorescent tracer. The relative contribution of NMDA receptors versus AMPA/kainate receptors to C-fibre-evoked excitatory postsynaptic currents of lamina I neurones significantly decreased with age between postnatal day 18 and 27, but was independent of the supraspinal projection of the neurones. We did not find a significant contribution of kainate receptors to C-fibre-evoked excitatory postsynaptic currents. Lamina I projection and unidentified neurones possessed functional GABAA and glycine receptors but received scarce action potential independent spontaneous GABAergic and glycinergic inhibitory input as measured by miniature inhibitory postsynaptic currents. The miniature excitatory postsynaptic current frequencies were five times higher in projection than in unidentified neurones. The predominance of excitatory synaptic input to projection neurones, taken together with the previous finding that their membranes are more easily excitable than those of unidentified neurones, may facilitate the induction of synaptic long-term potentiation. PMID- 15878939 TI - Gating of TRP channels: a voltage connection? AB - TRP channels represent the main pathways for cation influx in non-excitable cells. Although TRP channels were for a long time considered to be voltage independent, several TRP channels now appear to be weakly voltage dependent with an activation curve extending mainly into the non-physiological positive voltage range. In connection with this voltage dependence, there is now abundant evidence that physical stimuli, such as temperature (TRPV1, TRPM8, TRPV3), or the binding of various ligands (TRPV1, TRPV3, TRPM8, TRPM4), shift this voltage dependence towards physiologically relevant potentials, a mechanism that may represent the main functional hallmark of these TRP channels. This review discusses some features of voltage-dependent gating of TRPV1, TRPM4 and TRPM8. A thermodynamic principle is elaborated, which predicts that the small gating charge of TRP channels is a crucial factor for the large voltage shifts induced by various stimuli. Some structural considerations will be given indicating that, although the voltage sensor is not yet known, the C-terminus may substantially change the voltage dependence of these channels. PMID- 15878940 TI - The Ca2+ spark of mammalian muscle. Physiology or pathology? PMID- 15878941 TI - Both cGMP and peroxynitrite mediate chronic interleukin-6-induced negative inotropy in adult rat ventricular myocytes. AB - We previously showed that chronic exposure to interleukin (IL)-6 decreases contractile and sarcoplasmic reticular (SR) function assessed by postrest potentiation (PRP) via a nitric oxide (NO)-dependent mechanism in adult rat ventricular myocytes (ARVM). Cyclic GMP (cGMP) has been associated with NO associated negative inotropic effects of IL-6 during acute exposure; however, its role in chronic cardiac effects of IL-6 remains unclear. The present study examined the roles of cGMP and peroxynitrite (ONOO-) in chronic IL-6-induced negative inotropy in ARVM. After ARVM were exposed to IL-6 for 2-24 h, intracellular cGMP contents were time dependently increased; this was mimicked by a NO donor and abolished by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), an inhibitor of soluble guanylyl cyclase (sGC), or Rp-8-Br-cGMP, an inhibitor of cGMP-dependent protein kinase G (PKG). Meanwhile, the IL-6-induced decrease in PRP at 2 h was blocked by ODQ or Rp-8-Br-cGMP. By contrast, ODQ or Rp-8-Br-cGMP only attenuated the inhibition of PRP induced by IL-6 after 24 h exposure. Furthermore, IL-6 time dependently increased superoxide anion production and ONOO formation; the latter was abolished by 5,10,15,20-tetrakis-(4-sulphonatophenyl) porphyrinato iron (III) (FeTPPS), an ONOO- decomposition catalyst. Interestingly, FeTPPS had no effect on the IL-6-elicited decrease in PRP at 2 h, but attenuated it after 24 h exposure. Moreover, inhibition of sGC/cGMP/PKG, but not ONOO- formation, abolished the IL-6-induced inhibition of kinetics of myocyte contraction during 24 h exposure. We conclude that while the sGC/cGMP/PKG pathway was the primary mechanism for chronic IL-6-induced negative inotropy at 2 h, both sGC/cGMP/PKG and ONOO-, at least in part, mediate the IL-6-induced inhibition of SR function after 24 h exposure. PMID- 15878942 TI - Facilitating efferent inhibition of inner hair cells in the cochlea of the neonatal rat. AB - Cholinergic brainstem neurones make inhibitory synapses on outer hair cells (OHCs) in the mature mammalian cochlea and on inner hair cells (IHCs) prior to the onset of hearing. We used electrical stimulation in an excised organ of Corti preparation to examine evoked release of acetylcholine (ACh) onto neonatal IHCs from these efferent fibres. Whole-cell voltage-clamp recording revealed that low frequency (0.25-1 Hz) electrical stimulation produced evoked inhibitory postsynaptic currents (IPSCs) at a relatively high fraction of failures (65%) and with mean amplitudes of about -20 pA at -90 mV, corresponding to a quantum content of approximately 1. Evoked IPSCs had biphasic waveforms at -60 mV, were blocked reversibly by alpha-bungarotoxin and strychnine and are most likely mediated by the alpha9/alpha10 acetylcholine receptor, with subsequent activation of calcium-dependent potassium (SK2) channels. Paired pulse stimulation with intervals of 10-100 ms caused facilitation of 200-300% in the mean IPSC amplitude. A train of 10 pulses with an interpulse interval of 25 ms produced increasingly larger IPSCs with maximum amplitudes greater than -100 pA due to facilitation and summation throughout the train. Repetitive efferent stimulation at 5 Hz or higher hyperpolarized IHCs by 5-10 mV and could completely prevent the generation of calcium action potentials normally evoked by depolarizing current injection. PMID- 15878943 TI - Arterial baroreflex control of heart rate: determining factors and methods to assess its spontaneous modulation. PMID- 15878944 TI - Resetting of the arterial baroreflex increases orthostatic sympathetic activation and prevents postural hypotension in rabbits. AB - Since humans are under ceaseless orthostatic stress, the mechanism to maintain arterial pressure (AP) under orthostatic stress against gravitational fluid shift is of great importance. We hypothesized that (1) orthostatic stress resets the arterial baroreflex control of sympathetic nerve activity (SNA) to a higher SNA, and (2) resetting of the arterial baroreflex contributes to preventing postural hypotension. Renal SNA and AP were recorded in eight anaesthetized, vagotomized and aortic-denervated rabbits. Isolated intracarotid sinus pressure (CSP) was increased stepwise from 40 to 160 mmHg with increments of 20 mmHg (60 s for each CSP level) while the animal was placed supine and at 60 deg upright tilt. Upright tilt shifted the CSP-SNA relationship (the baroreflex neural arc) to a higher SNA, shifted the SNA-AP relationship (the baroreflex peripheral arc) to a lower AP, and consequently moved the operating point to marked high SNA while maintaining AP. A simulation study suggests that resetting in the neural arc would double the orthostatic activation of SNA and increase the operating AP in upright tilt by 10 mmHg, compared with the absence of resetting. In addition, upright tilt did not change the CSP-AP relationship (the baroreflex total arc). A simulation study suggests that although a downward shift of the peripheral arc could shift the total arc downward, resetting in the neural arc would compensate this fall and prevent the total arc from shifting downward to a lower AP. In conclusion, upright tilt increases SNA by resetting the baroreflex neural arc. This resetting may compensate for the reduced pressor responses to SNA in the peripheral cardiovascular system and contribute to preventing postural hypotension. PMID- 15878945 TI - Hypoxic activation of arterial chemoreceptors inhibits sympathetic outflow to brown adipose tissue in rats. AB - In urethane-chloralose anaesthetized, neuromuscularly blocked, artificially ventilated rats, we demonstrated that activation of carotid chemoreceptors inhibits the elevated levels of brown adipose tissue (BAT) sympathetic nerve activity (SNA) evoked by hypothermia, by microinjection of prostaglandin E2 into the medial preoptic area or by disinhibition of neurones in the raphe pallidus area (RPa). Peripheral chemoreceptor stimulation with systemic administration of NaCN (50 microg in 0.1 ml) or with hypoxic ventilation (8% O2-92% N2, 30 s) completely inhibited BAT SNA. Arterial chemoreceptor-evoked inhibition of BAT SNA was eliminated by prior bilateral transections of the carotid sinus nerves or by prior inhibition of neurones within the commissural nucleus tractus solitarii (commNTS) with glycine (40 nmol/80 nl) or with the GABAA receptor agonist muscimol (160 pmol/80 nl; 77 +/- 10% attenuation), or by prior blockade of ionotropic excitatory amino acid receptors in the commNTS with kynurenate (8 nmol/80 nl; 82 +/- 10% attenuation). Furthermore, activation of commNTS neurones following local microinjection of bicuculline (30 pmol/60 nl) completely inhibited the elevated level of BAT SNA resulting from disinhibition of neurones in the RPa. These results demonstrate that hypoxic stimulation of arterial chemoreceptor afferents leads to an inhibition of BAT SNA and BAT thermogenesis through an EAA-mediated activation of second-order, arterial chemoreceptor neurones in the commNTS. Peripheral chemoreceptor-evoked inhibition of BAT SNA could directly contribute to (or be permissive for) the hypoxia-evoked reductions in body temperature and oxygen consumption that serve as an adaptive response to decreased oxygen availability. PMID- 15878946 TI - Gain modulation by serotonin in pyramidal neurones of the rat prefrontal cortex. AB - Serotonin (5-HT) is widely implicated in brain functions and diseases. The vertebrate brain is extensively innervated by 5-HT fibres originating from the brain stem, and 5-HT axon terminals interact with other neurones in complex ways. The cellular mechanisms underlying 5-HT function in the brain are not well understood. The present study examined the effect of 5-HT on the responsiveness of neurones in the neocortex. Using patch-clamp recording in acute slices, we showed that 5-HT substantially increased the slope (gain) of the firing rate current curve in layer 5 pyramidal neurones of the rat prefrontal cortex. The effect of 5-HT on gain is confined to the range of firing rate (0-10 Hz) that is known to be behaviourally relevant. 5-HT also changed current threshold for spike train generation, but this effect was inconsistent, and was independent of the effect on gain. The gain modulation by 5-HT was mediated by 5-HT2 receptors, and involved postsynaptic mechanisms. 5-HT2-mediated gain increase could not be attributed to changes in the membrane potential, the input resistance or the properties of action potentials, but was associated with a reduction of the afterhyperpolarization and an induction of the slow afterdepolarization. Blocking Ca2+ entry with Cd2+ increased the gain by itself and blocked 5-HT2- mediated gain increase. Buffering [Ca2+](i) with 25 mM EGTA also substantially reduced 5 HT2- mediated gain increase. Noradrenaline, which blocked the afterhyperpolarization, also induced a moderate increase in gain. Together, our results suggest that 5-HT may regulate the dynamics of cortical circuits through multiplicative scaling. PMID- 15878947 TI - Mechanisms underlying the early phase of spike frequency adaptation in mouse spinal motoneurones. AB - Spike frequency adaptation (SFA) is a fundamental property of repetitive firing in motoneurones (MNs). Early SFA (occurring over several hundred milliseconds) is thought to be important in the initiation of muscular contraction. To date the mechanisms underlying SFA in spinal MNs remain unclear. In the present study, we used both whole-cell patch-clamp recordings of MNs in lumbar spinal cord slices prepared from motor functionally mature mice and computer modelling of spinal MNs to investigate the mechanisms underlying SFA. Pharmacological blocking agents applied during whole-cell recordings in current-clamp mode demonstrated that the medium AHP conductance (apamin), BK-type Ca2+ -dependent K+ channels (iberiotoxin), voltage-activated Ca2+ channels (CdCl2), M-current (linopirdine) and persistent Na+ currents (riluzole) are all unnecessary for SFA. Measurements of Na+ channel availability including action potential amplitude, action potential threshold and maximum depolarization rate of the action potential were found to correlate with instantaneous firing frequency suggesting that the availability of fast, inactivating Na+ channels is involved in SFA. Characterization of this Na+ conductance in voltage-clamp mode demonstrated that it undergoes slow inactivation with a time course similar to that of SFA. When experimentally measured parameters for the fast, inactivating Na+ conductance (including slow inactivation) were incorporated into a MN model, SFA could be faithfully reproduced. The removal of slow inactivation from this model was sufficient to remove SFA. These data indicate that slow inactivation of the fast, inactivating Na+ conductance is likely to be the key mechanism underlying early SFA in spinal MNs. PMID- 15878948 TI - Enhancement of calcium-dependent afterpotentials in oxytocin neurons of the rat supraoptic nucleus during lactation. AB - The firing pattern of oxytocin (OT) hormone synthesizing neurons changes dramatically immediately before each milk ejection, when a brief burst of action potentials is discharged. OT neurons possess intrinsic currents that would modulate this burst. Our previous studies showed the amplitude of the Ca2+ dependent afterhyperpolarization (AHP) following spike trains is significantly larger during lactation. In the present study we sought to determine which component of the AHP is enhanced, and whether the enhancement could be related to changes in whole-cell Ca2+ current or the Ca2+ transient in identified OT or vasopressin (VP) neurons during lactation. We confirmed, with whole-cell current clamp recordings, our previous finding from sharp electrodes that the size of the AHP following spike trains increased in OT, but not VP neurons during lactation. We then determined that an apamin-sensitive medium-duration AHP (mAHP) and an apamin-insensitive slow AHP (sAHP) were specifically increased in OT neurons. Simultaneous Ca2+ imaging revealed that the peak change in somatic [Ca2+]i was not altered in either cell type, but the slow decay of the Ca2+ transient was faster in both cell types during lactation. In voltage clamp, the whole-cell, Ca2+ current was slightly larger during lactation in OT cells only, but current density was unchanged when corrected for somatic hypertrophy. The currents, ImAHP and IsAHP, also were increased in OT neurons only, but only the apamin-sensitive ImAHP showed an increase in current density after adjusting for somatic hypertrophy. These findings suggest a specific modulation (e.g. increased number) of the small-conductance Ca2+ -dependent K+ (SK) channels, or their interaction with Ca2+, underlies the increased mAHP/ImAHP during lactation. This larger mAHP may be necessary to limit the explosive bursts during milk ejection. PMID- 15878949 TI - Mechanism of riboflavin uptake by cultured human retinal pigment epithelial ARPE 19 cells: possible regulation by an intracellular Ca2+-calmodulin-mediated pathway. AB - In mammalian cells (including those of the ocular system), the water-soluble vitamin B2 (riboflavin, RF) assumes an essential role in a variety of metabolic reactions and is critical for normal cellular functions, growth and development. Cells of the human retinal pigment epithelium (hRPE) play an important role in providing a sufficient supply of RF to the retina, but nothing is known about the mechanism of the vitamin uptake by these cells and its regulation. Our aim in the present study was to address this issue using the hRPE ARPE-19 cells as the retinal epithelial model. Our results show RF uptake in the hRPE to be: (1) energy and temperature dependent and occurring without metabolic alteration in the transported substrate, (2) pH but not Na+ dependent, (3) saturable as a function of concentration with an apparent Km of 80 +/- 14 nM, (4) trans stimulated by unlabelled RF and its structural analogue lumiflavine, (5) cis inhibited by the RF structural analogues lumiflavine and lumichrome but not by unrelated compounds, and (6) inhibited by the anion transport inhibitors 4,4' diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) and 4-acetamido-4' isothiocyanatostilbene-2,2'-disulphonic acid (SITS) as well as by the Na+ -H+ exchange inhibitor amiloride and the sulfhydryl group inhibitor p chloromercuriphenylsulphonate (p-CMPS). Maintaining the hRPE cells in a RF deficient medium led to a specific and significant up-regulation in RF uptake which was mediated via changes in the number and affinity of the RF uptake carriers. While modulating the activities of intracellular protein kinase A (PKA) , protein kinase C (PKC)-, protein tyrosine kinase (PTK)-, and nitric oxide (NO) mediated pathways were found to have no role in regulating RF uptake, a role for the Ca2+ -calmodulin-mediated pathway was observed. These studies demonstrate for the first time the involvement of a specialized carrier-mediated mechanism for RF uptake by hRPE cells and show that the process is adaptively regulated in RF deficiency, and also appears to be under the regulation of an intracellular Ca2+ calmodulin-mediated pathway. PMID- 15878950 TI - Intestinal gluconeogenesis and glucose transport according to body fuel availability in rats. AB - Intestinal hexose absorption and gluconeogenesis have been studied in relation to refeeding after two different fasting phases: a long period of protein sparing during which energy expenditure is derived from lipid oxidation (phase II), and a later phase characterized by a rise in plasma corticosterone triggering protein catabolism (phase III). Such a switch in body fuel uses, leading to changes in body reserves and gluconeogenic precursors, could modulate intestinal gluconeogenesis and glucose transport. The gene and protein levels, and the cellular localization of the sodium-glucose cotransporter SGLT1, and of GLUT5 and GLUT2, as well as that of the key gluconeogenic enzymes phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (Glc6Pase) were measured. PEPCK and Glc6Pase activities were also determined. In phase III fasted rats, SGLT1 was up-regulated and intestinal glucose uptake rates were higher than in phase II fasted and fed rats. PEPCK and Glc6Pase mRNA, protein levels and activities also increased in phase III. GLUT5 and GLUT2 were down-regulated throughout the fast, but increased after refeeding, with GLUT2 recruited to the apical membrane. The increase in SGLT1 expression during phase III may allow glucose absorption at low concentrations as soon as food is available. Furthermore, an increased epithelial permeability due to fasting may induce a paracellular movement of glucose. In the absence of intestinal GLUT2 during fasting, Glc6Pase could be involved in glucose release to the bloodstream via membrane trafficking. Finally, refeeding triggered GLUT2 and GLUT5 synthesis and apical recruitment of GLUT2, to absorb larger amounts of hexoses. PMID- 15878951 TI - Hexokinase II protein content is a determinant of exercise endurance capacity in the mouse. AB - Hexokinase (HK) II content is elevated in fatigue resistant muscle fibres and exercise trained muscle. The aim of this study was to determine if exercise capacity is dependent on muscle HK protein content. C57Bl/6 mice with a 50% HK knockout (HK+/-), no genetic manipulation (wild-type, WT) and an approximately 3 fold HK overexpression (HKTg) were tested. Mice (n = 12/group) completed both a maximal oxygen consumption test(VO2max) test and an endurance capacity test (run at approximately 75% VO2max) on an enclosed treadmill equipped to measure gas exchange. Arterial and venous catheters were surgically implanted into separate groups of mice (n = 9-11/group) in order to measure an index of muscle glucose uptake Rg during 30 min of treadmill exercise. Maximum work rate (0.95 +/- 0.05, 1.00 +/- 0.04 and 1.06 +/- 0.07 kg m min-1), (137 +/- 3, 141 +/- 4 and 141 +/- 5 ml kg-1 min-1) and maximal respiratory exchange ratio (1.04 +/- 0.02, 1.00 +/- 0.03 and 1.04 +/- 0.04) were similar in HK+/-, WT and HKTg, respectively. Exercise endurance capacity (measured as time to exhaustion) increased as HK content increased (55 +/- 11, 77 +/- 5 and 98 +/- 9 min) and this was related to Rg measured in mice during 30 min of exercise (13 +/- 2, 24 +/- 5 and 42 +/- 5 micromol (100 g)-1 min-1). Muscle glycogen in sedentary HK+/-mice and HK+/- mice following 30 min of exercise were significantly lower than in HKTg and WT mice. However, the net exercise-induced muscle glycogen breakdown was equal in the three genotypes. In summary, HK protein content within the range studied (a) was not associated with a difference in the capacity to perform maximal intensity exercise, (b) was a powerful determinant of the ability to sustain moderate intensity exercise, as reducing HK content impaired endurance and increasing HK content enhanced endurance, and (c) although directly related to exercise endurance, was not a determinant of net muscle glycogen usage during exercise. In conclusion, adaptations that increase HK protein content and/or functional activity such as regular exercise contribute to increased muscular endurance. PMID- 15878952 TI - Maternal nutrient deprivation induces sex-specific changes in thyroid hormone receptor and deiodinase expression in the fetal guinea pig brain. AB - Thyroid hormone deprivation during fetal life has been implicated in neurodevelopmental morbidity. In humans, poor growth in utero is also associated with fetal hypothyroxinaemia. In guinea pigs, a short period (48 h) of maternal nutrient deprivation at gestational day (gd) 50 results in fetuses with hypothyroxinaemia and increased brain/body weight ratios. Thyroid hormone action is mediated by nuclear thyroid hormone receptors (TRs) and is dependent upon the prereceptor regulation of supply of triiodothyronine (T3) by deiodinase enzymes. Examination of fetal guinea pig brains using in situ hybridization demonstrated widespread expression of mRNAs encoding TRalpha1, alpha2 and beta1, with regional colocalization of deiodinase type 2 (D2) mRNA in the developing forebrain, limbic structures, brainstem and cerebellum at gd52. With maternal nutrient deprivation, TRalpha1 and beta1 mRNA expression was significantly increased in the male, but decreased in the female fetal hippocampus and cerebellum and other areas showing high TR expression under euthyroid conditions. Maternal nutrient deprivation resulted in elevated D2 mRNA expression in males and females. Deiodinase type 3 (D3) mRNA expression was confined to the shell of the nucleus accumbens, the posterior amygdalohippocampal area, brainstem and cerebellum, and did not change with maternal nutrient deprivation. In conclusion, maternal nutrient deprivation resulted in sex-specific changes in TR mRNA expression and a generalized increase in D2 mRNAs within the fetal brain. These changes may represent a protective mechanism to maintain appropriate thyroid hormone action in the face of fetal hypothyroxinaemia in order to optimize brain development. PMID- 15878953 TI - Medullary serotonergic neurones modulate the ventilatory response to hypercapnia, but not hypoxia in conscious rats. AB - Serotonergic neurones in the mammalian medullary raphe region (MRR) have been implicated in central chemoreception and the modulation of the ventilatory response to hypercapnia, and may also be involved in the ventilatory response to hypoxia. In this study, we ask whether ventilatory responses across arousal states are affected when the 5-hydroxytryptamine 1A receptor (5-HT1A) agonist (R) (+)-8-hydroxy-2(di-n-propylamino)tetralin (DPAT) is microdialysed into the MRR of the unanaesthetized adult rat. Microdialysis of 1, 10 and 30 mM DPAT into the MRR significantly decreased absolute ventilation values(VE) during 7% CO2 breathing by 21%, 19% and 30%, respectively, in wakefulness compared to artificial cerebrospinal fluid (aCSF) microdialysis, due to decreases in tidal volume (VT) and not in frequency (f), similar to what occurred during non-rapid eye movement (NREM) sleep. The concentration-dependence of the hypercapnic ventilatory effect might be due to differences in tissue distribution of DPAT. DPAT (30 mM) changed room air breathing pattern by increasing f and decreasing VT. As evidenced by a sham control group, repeated experimentation and microdialysis of aCSF alone had no effect on the ventilatory response to 7% CO2 during wakefulness or sleep. Unlike during hypercapnia, microdialysis of 30 mM DPAT into the MRR did not change the ventilatory response to 10% O2. Additionally, 10 and 30 mM DPAT MRR microdialysis decreased body temperature, and 30 mM DPAT increased the percentage of experimental time in wakefulness. We conclude that serotonergic activity in the MRR plays a role in the ventilatory response to hypercapnia, but not to hypoxia, and that MRR 5-HT1A receptors are also involved in thermoregulation and arousal. PMID- 15878954 TI - Computing with thalamocortical ensembles during different behavioural states. AB - A series of recent studies have indicated that ensembles of neurones, distributed within the neural structures that form the primary thalamocortical loop (TCL) of the trigeminal component of the rat somatosensory system, change the way they respond to similar tactile stimuli, according to both the behavioural strategy employed by animals to gather information and the animal's internal brain states. These findings suggest that top-down influences, which are more likely to play a role during active discrimination than during passive whisker stimulation, may alter the pattern of neuronal firing within both the distinct layers of the primary somatosensory cortex (S1) and the ventral posterior medial nucleus (VPM). We propose that through this physiological process, which involves concurrent dynamic modulations at both cellular and circuit levels in the TCL, rats can either optimize the detection of novel or hard to sense stimuli or they can analyse complex patterns of multi-whisker stimulation, during natural exploration of their surrounding environment. PMID- 15878955 TI - General receptor for phosphoinositides 1, a novel repressor of thyroid hormone receptor action that prevents deoxyribonucleic acid binding. AB - Thyroid hormone receptors (TRs) bind to response elements (TREs) located in the promoter region of target genes and modulate their transcription. The effects of TRs require the presence of coregulators that act as adaptor molecules between TRs and complexes that are involved in chromatin remodeling or that directly contact the basal transcription machinery. Using the yeast two-hybrid system, we identified a new interacting partner for TRs: GRP1 (general receptor for phosphoinositides-1), a nucleotide exchange factor, which had never been shown to interact with nuclear receptors. We reconfirmed the interaction between TRs and GRP1 in yeast and glutathione-S-transferase pull-down assays, and determined the areas of TRs and GRP1 involved in the interaction. Coimmunoprecipitation studies demonstrated that the interaction between GRP1 and TRs takes place in the cytoplasm and the nucleus of mammalian cells. To assess functional consequences of the interaction, we used transient transfection of CV-1 cells with TR and GRP1 expression vectors and luciferase reporter genes. On positive TREs, GRP1 decreased activation by 45-60%. On the negative TREs it increased repression by blunting the activation in the absence of T3, except for TRbeta2, which was not affected. Using EMSA, we have determined that addition of GRP1 diminishes the formation of TR/TR homodimers and TR/retinoid X receptor heterodimers on TREs, which could explain the effect of GRP1 on transcription. Furthermore, protein interaction assays using increasing concentrations of double-stranded TREs show a dose-dependent decrease of the interaction between GRP1 and TRs. The homo/heterodimers formed by TRs and retinoic X receptor-alpha were not influenced by the presence of GRP1, also suggesting that GRP1 interferes directly with DNA binding. Taken together, these data provide evidence that GRP1 is a new corepressor for TRs, which modulates both positive and negative regulation by T3 by decreasing TR-complex formation on TREs. PMID- 15878956 TI - Zero-length cross-linking reveals that tight interactions between the extracellular and transmembrane domains of the luteinizing hormone receptor persist during receptor activation. AB - Several molecular models of glycoprotein hormone receptor activation have been proposed. It has been suggested that ligand binding to the ectodomain (ECD) leads to major changes in intramolecular interactions between the ECD and the transmembrane domain. We studied these intramolecular modifications by generating a recombinant LH/CG receptor (LHR) bearing an intramolecular cleavage site. We did this by inserting a furin site at position 316 in the hinge region of the ECD (LHR_Fur316). Affinity for human chorionic gonadotropin (hCG) and cAMP production upon hCG stimulation was identical to those of wild-type LHR. Western blot analysis showed that the LHR_Fur316 receptor was cleaved into two subunits linked by disulfide bridges. Chemical shedding of the ECD from the transmembrane domain did not increase basal adenylate cyclase activity, indicating that the first 294 residues did not act as an inverse agonist. The truncated LHR_316 was still activated by hCG but with an EC50 higher than that for the wild-type receptor. Zero length cross-linking was used to study intramolecular interactions between the two domains of LHR_Fur316. Cross-linking efficiency was similar for the basal and activated states, which indicated that the two domains interacted closely in the basal state, and this tight interaction persisted during activation. Our data suggest that activation of the LHR results from subtle modifications of intramolecular interactions between the two domains and low-affinity binding of hCG to the extracellular loops or residues preceding the first transmembrane segment. PMID- 15878957 TI - Constitutively active Gq impairs gonadotropin-releasing hormone-induced intracellular signaling and luteinizing hormone secretion in LbetaT2 cells. AB - Chronic GnRH treatment causes homologous desensitization by reducing GnRH receptor and Gq/11 expression and by down-regulating protein kinase C (PKC), cAMP, and calcium-dependent signaling. It also causes heterologous desensitization of other Gq-coupled receptors, but the mechanisms involved remain elusive. In this study, we investigated the effect of constitutive activation of Gq signaling on GnRH-induced signaling and LH secretion. We show that adenoviral expression of a constitutively active mutant Gq(Q209L) results in a state of GnRH resistance but does not alter GnRH receptor expression. We observed that Gq(Q209L) reduced expression of phospholipase C (PLC)beta1, a target of Gq in these cells, but not PLCbeta3 or PLCgamma1. Downstream of PLCbeta1, expression of novel PKC isoforms (delta and epsilon) was reduced. Adenoviral expression of a kinase-inactive, dominant-negative version of PKCdelta impaired GnRH activation of ERK, but not induction of c-Fos and LHbeta proteins, indicating that the novel PKCs signal to the ERK cascade. Despite reductions in PLCbeta1, calcium responses to GnRH were elevated in Gq(Q209L)-infected cells due to increased calcium influx through L-type calcium channels. Paradoxically, downstream calcium-dependent signaling and LH secretion were impaired. Taken together, these data demonstrate that prolonged activation of the Gq pathway desensitizes GnRH-induced signaling by selectively down-regulating the PLC-PKC-Ca2+ pathway, leading to reduced LHbeta synthesis and LH secretion. PMID- 15878958 TI - Myostatin inhibits myogenesis and promotes adipogenesis in C3H 10T(1/2) mesenchymal multipotent cells. AB - Inactivating mutations of the mammalian myostatin gene are associated with increased muscle mass and decreased fat mass; conversely, myostatin transgenic mice that overexpress myostatin in the skeletal muscle have decreased muscle mass and increased fat mass. We investigated the effects of recombinant myostatin protein and antimyostatin antibody on myogenic and adipogenic differentiation of mesenchymal multipotent cells. Accordingly, 10T(1/2) cells were incubated with 5' azacytidine for 3 d to induce differentiation and then treated with a recombinant protein for myostatin (Mst) carboxy terminal 113 amino acids or a polyclonal anti Mst antibody for 3, 7, and 14 d. Cells were also cotransfected with a Mst cDNA plasmid expressing the full-length 375-amino acid protein (pcDNA-Mst375) and the silencer RNAs for either Mst (pSil-Mst) or a random sequence (pSil-RS) for 3 or 7 d, and Mst expression was determined. Adipogenesis was evaluated by quantitative image analysis of fat cells before and after oil-red-O staining, immunocytochemistry of adiponectin, and Western blot for CCAAT/enhancer binding protein-alpha. Myogenesis was estimated by quantitative image analysis immunocytochemistry for MyoD (Myo differentiation protein), myogenin, and myosin heavy chain type II, or by Western blot for myogenin. 5'-Azacytidine-mediated differentiation induced endogenous full-length Mst expression. Recombinant Mst carboxy terminal 113 amino acids inhibited both early and late markers of myogenesis and stimulated both early and late markers of adipogenesis, whereas the antibody against Mst exerted the reverse effects. Myogenin levels at 7 d after transfection of pcDNA-Mst375 were reduced as expected and elevated by pSil Mst, which blocked efficiently Mst375 expression. In conclusion, myostatin promotes the differentiation of multipotent mesenchymal cells into the adipogenic lineage and inhibits myogenesis. PMID- 15878959 TI - Heat shock protein-70 expressed on the surface of cancer cells binds parathyroid hormone-related protein in vitro. AB - Recent studies have shown that the functions of PTH-related protein (PTHrP) and its derived peptides cannot be attributed solely to PTH/PTHrP receptor binding. The present study focused on the identification of other proteins that might bind PTHrP at the cell surface. Using affinity chromatography, we applied extracts of cell-surface biotinylated proteins from cancer and normal cell lines over Sepharose beads coupled with different PTHrP-derived peptides. Elution with the corresponding free peptide revealed a major protein of about 70 kDa that was present in all of the PTHrP peptide eluates from cancer cell extracts but not from normal breast cell extracts. Mass spectroscopy analysis and immunoblotting identified this PTHrP-binding protein as heat shock protein-70 (HSP70). Using a recently published algorithm that predicts HSP70 binding sites within proteins, we found that all four PTHrP peptides used in these studies contain amino acid motifs with high probabilities for HSP70 binding in vivo. Cell culture studies in the presence of a polyclonal anti-HSP70 antibody demonstrated increased PTHrP secretion, decreased total cellular protein, and differentially regulated proliferation. Taken together, these studies demonstrate a novel and biologically relevant interaction between cell surface-expressed HSP70 and PTHrP in cancer. PMID- 15878960 TI - Essential role of the oocyte in estrogen amplification of follicle-stimulating hormone signaling in granulosa cells. AB - The establishment of dominant ovarian follicles that are capable of ovulating fertilizable oocytes is a fundamental determinant of female fertility. This process is governed by pituitary gonadotropins as well as local ovarian factors. Within the follicle, estrogen acts in an autocrine/paracrine manner to enhance FSH action in the granulosa cells. These effects include the augmentation of P450aromatase expression and estradiol production. This feed-forward effect of estrogen is believed to play a key role in follicle dominance. Here we found the essential role of the oocyte in this physiological process using primary cultures of rat granulosa cells. In the presence, but not absence, of oocytes, estrogen amplified FSH-stimulated increases in mRNA expression of P450aromatase, FSH receptor, LH receptor, and inhibin alpha-, betaA-, and betaB-subunits as well as cAMP production. Thus, oocytes mediate the estrogen enhancement of FSH action in the granulosa cells. In comparison with FSH, cotreatment with estrogen and oocytes failed to amplify the stimulatory effects of forskolin or 8 bromoadenosine-cAMP on granulosa cell responses including P450aromatase mRNA expression and cAMP production, indicating that estrogen/oocytes amplify FSH action at a site upstream of adenylate cyclase. These findings support the novel conclusion that communication between the oocyte and granulosa cells plays a crucial role in mediating estrogen action during FSH-dependent folliculogenesis. PMID- 15878961 TI - Progesterone receptor isoforms A and B: temporal and spatial differences in expression during murine mammary gland development. AB - Progesterone is a potent mitogen in the mammary gland. Based on studies using cells and animals engineered to express progesterone receptor (PR) isoforms A or B, PRA and PRB are believed to have different functions. Using an immunohistochemical approach with antibodies specific for PRA only or PRB only, we show that PRA and PRB expression in mammary epithelial cells is temporally and spatially separated during normal mammary gland development in the BALB/c mouse. In the virgin mammary gland when ductal development is active, the only PR protein isoform expressed was PRA. PRA levels were significantly lower during pregnancy, suggesting a minor role at this stage of development. PRB was abundantly expressed only during pregnancy, during alveologenesis. PRA and PRB colocalization occurred in only a small percentage of cells. During pregnancy there was extensive colocalization of PRB with 5-bromo-2'-deoxyuridine (BrdU) and cyclin D1; 95% of BrdU-positive cells and 83% of cyclin D1-positive cells expressed PRB. No colocalization of PRA with either BrdU or cyclin D1 was observed at pregnancy. In the virgin gland, PRA colocalization with BrdU or cyclin D1 was low; only 27% of BrdU-positive cells and 4% of cyclin D1-positive cells expressed PRA. The implication of these findings is that different actions of progesterone are mediated in PRB positive vs. PRA-positive cells in vivo. The spatial and temporal separation of PR isoform expression in mouse mammary gland provides a unique opportunity to determine the specific functions of PRA vs. PRB in vivo. PMID- 15878962 TI - Hedgehog signaling in mouse ovary: Indian hedgehog and desert hedgehog from granulosa cells induce target gene expression in developing theca cells. AB - Follicle development in the mammalian ovary requires interactions among the oocyte, granulosa cells, and theca cells, coordinating gametogenesis and steroidogenesis. Here we show that granulosa cells of growing follicles in mouse ovary act as a source of hedgehog signaling. Expression of Indian hedgehog and desert hedgehog mRNAs initiates in granulosa cells at the primary follicle stage, and we find induced expression of the hedgehog target genes Ptch1 and Gli1, in the surrounding pre-theca cell compartment. Cyclopamine, a highly specific hedgehog signaling antagonist, inhibits this induced expression of target genes in cultured neonatal mouse ovaries. The theca cell compartment remains a target of hedgehog signaling throughout follicle development, showing induced expression of the hedgehog target genes Ptch1, Ptch2, Hip1, and Gli1. In periovulatory follicles, a dynamic synchrony between loss of hedgehog expression and loss of induced target gene expression is observed. Oocytes are unable to respond to hedgehog because they lack expression of the essential signal transducer Smo (smoothened). The present results point to a prominent role of hedgehog signaling in the communication between granulosa cells and developing theca cells. PMID- 15878963 TI - Cloning and characterization of a functional type II gonadotropin-releasing hormone receptor with a lengthy carboxy-terminal tail from an ancestral vertebrate, the sea lamprey. AB - A full-length transcript encoding a functional type II GnRH receptor was cloned from the pituitary of the sea lamprey, Petromyzon marinus. The current study is the first to identify a pituitary GnRH receptor transcript in an agnathan, which is the oldest vertebrate lineage. The cloned receptor retains the conserved structural features and amino acid motifs of other known GnRH receptors and notably includes a C-terminal intracellular tail of approximately 120 amino acids, the longest C-terminal tail of any vertebrate GnRH receptor identified to date. The lamprey GnRH receptor was shown to activate the inositol phosphate (IP) signaling system; stimulation with either lamprey GnRH-I or lamprey GnRH-III led to dose-dependent responses in transiently transfected COS7 cells. Furthermore, analyses of serially truncated lamprey GnRH receptor mutants indicate perturbations of the C-terminal tail disrupts IP accumulation, however, the tailless lamprey GnRH receptor was not only functional but was also capable of stimulating IP levels equal to wild type. Expression of the receptor transcript was demonstrated in the pituitary and testes using RT-PCR, whereas in situ hybridization showed expression and localization of the transcript in the proximal pars distalis of the pituitary. The phylogenetic placement and structural and functional features of this GnRH receptor suggest that it is representative of an ancestral GnRH receptor. In addition to having an important role in lamprey reproductive processes, the extensive C-terminal tail of this lamprey GnRH receptor may have great significance for understanding the evolutionary change of this vital structural feature within the GnRH receptor family. PMID- 15878964 TI - Deficiency of the very low-density lipoprotein (VLDL) receptors in streptozotocin induced diabetic rats: insulin dependency of the VLDL receptor. AB - Hyperlipidemia is a common feature of diabetes and is related to cardiovascular disease. The very low-density lipoprotein receptor (VLDL-R) is a member of the low-density lipoprotein receptor (LDL-R) family. It binds and internalizes triglyceride-rich lipoproteins with high specificity. We examined the etiology of hyperlipidemia in the insulin-deficient state. VLDL-R expression in heart and skeletal muscle were measured in rats with streptozotocin (STZ)-induced diabetes. STZ rats showed severe hyperlipidemia on d 21 and 28, with a dramatic decline in VLDL-R protein in skeletal muscle (>90%), heart (approximately 50%) and a loss of adipose tissues itself on d 28. The reduction of VLDL-R protein in skeletal muscle could not be explained simply by a decrease at the transcriptional level, because a dissociation between VLDL-R protein and mRNA expression was observed. The expression of LDL-R and LDL-R-related protein in liver showed no consistent changes. Furthermore, no effect on VLDL-triglyceride production in liver was observed in STZ rats. A decrease in postheparin plasma lipoprotein lipase activity started on d 7 and continued to d 28 at the 50% level even though severe hyperlipidemia was detected only on d 21 and 28. In rat myoblast cells, serum deprivation for 24 h induced a reduction in VLDL-R proteins. Insulin (10(-6) m), but not IGF-I (10 ng/ml), restored the decreased VLDL-R proteins by serum deprivation. These results suggest that the combination of VLDL-R deficiency and reduced plasma lipoprotein lipase activity may be responsible for severe hyperlipidemia in insulin-deficient diabetes. PMID- 15878965 TI - Attenuation of diet-induced weight gain and adiposity through increased energy expenditure in mice lacking angiotensin II type 1a receptor. AB - Given that angiotensin II (AII) type 1 and 2 receptors (Agtr1 and Agtr2) are expressed in adipose tissue, AII may act directly on adipose tissue. However, regardless of whether AII directly modulates adipose tissue growth and metabolism in vivo and, if so, whether it is mediated via Agtr1 are still matters of debate. To understand the functional role of Agtr1 in adipose tissue growth and metabolism in vivo, we examined the metabolic phenotypes of mice lacking Agtr1a (Agtr1a-/- mice) during a high-fat diet. The Agtr1a-/- mice exhibited the attenuation of diet-induced body weight gain and adiposity, and insulin resistance relative to wild-type littermates (Agtr1a+/+ mice). They also showed increased energy expenditure accompanied by sympathetic activation, as revealed by increased rectal temperature and oxygen consumption, increased expression of uncoupling protein-1 mRNA in brown adipose tissue, and increased urinary catecholamine excretion. The heterozygous Agtr1a-deficient mice (Agtr1a+/- mice) also exhibited metabolic phenotypes similar to those of Agtr1a-/- mice. Using mouse embryonic fibroblasts derived from Agtr1a+/+ and Agtr1a-/- mice, we found no significant difference between genotypes in the ability to differentiate into lipid-laden mature adipocytes. In primary cultures of mouse mature adipocytes, AII increased the expression of mRNAs for some adipocytokines, which was abolished by pharmacological blockade of Agtr1. This study demonstrates that Agtr1a-/- mice exhibit attenuation of diet-induced weight gain and adiposity through increased energy expenditure. The data also suggest that AII does not affect directly adipocyte differentiation, but can modulate adipocytokine production via Agtr1. PMID- 15878966 TI - Effect of follicle-stimulating hormone and estrogen on the expression of betaglycan messenger ribonucleic acid levels in cultured rat granulosa cells. AB - Betaglycan (TGFbeta type III receptor) was recently identified as a coreceptor to enhance the binding of inhibin A to activin type II receptor. This inhibin/betaglycan/activin type II receptor complex prevents activins from binding to their own receptors. The present study was undertaken to identify the expression and the regulation of the betaglycan gene in cultured rat granulosa cells. Northern blot analysis indicated betaglycan mRNA transcript of approximately 6.4 kbp. The treatment of the cells with FSH increased the betaglycan mRNA level, and a concurrent treatment with estradiol brought a significant increase in betaglycan mRNA. The protein kinase A activator, 8 bromoadenosine-cAMP, also increased the expression of its mRNA. Furthermore, betaglycan mRNA was induced additively by estradiol, which was blocked by estrogen receptor antagonists [ICI 182780, (R, R)-cis-diethyltetrahydro-2,8 chrysenediol]. In the luciferase assay, FSH altered the promoter activity of betaglycan. Moreover, when FSH plus estradiol was added to the granulosa cells, a significant increase in the half-life of betaglycan mRNA transcript was seen. In summary, FSH and estradiol increased betaglycan mRNA expression, most possibly through the protein kinase A pathway and the estrogen receptor-beta. The increase of betaglycan mRNA was due to an increase in transcription and altered mRNA stability. In ovarian regulatory function, the expression of betaglycan may involve the functional antagonism of inhibin A in activin signal transduction. PMID- 15878967 TI - Lysophospholipids enhance matrix metalloproteinase-2 expression in human endothelial cells. AB - Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are both low molecular-weight lysophospholipids, which promote cell proliferation, migration, and invasion via interaction with a family of specific G protein-coupled receptors. Matrix metalloproteinases (MMPs) are zinc-dependent proteolytic enzymes, which are involved in degradation of the extracellular matrix and play critical roles in endothelial cell migration and matrix remodeling during angiogenesis. Among these MMPs, MMP-2 is known to trigger cell migration. In our present study, we examined the effects of LPA and S1P on MMP-2 expression in human endothelial cells. We showed that LPA and S1P enhanced MMP-2 expression in mRNA, protein levels, and also enzymatic activity of cells of the EAhy926 human endothelial cell line. The enhancement effects occurred in concentration- and time-dependent manners. Results from real-time PCR, Western blots, and substrate gels indicated that these enhancement effects were mediated through MAPK kinase/ERK-, nuclear factor-kappaB-, and calcium influx-dependent pathways. Furthermore, we show that endothelial cell invasion of the gel was enhanced by lysophospholipids, and the induction could be prevented by an MMP inhibitor, GM6001. These observations suggest that LPA and S1P may play important roles in endothelial cell invasion by regulating the expression of MMP-2. PMID- 15878968 TI - Transcriptional regulation of dehydroepiandrosterone sulfotransferase (SULT2A1) by estrogen-related receptor alpha. AB - The estrogen-related receptors (ERRalpha, -beta, and -gamma) are a subfamily of orphan nuclear receptors (designated NR3B1, NR3B2, and NR3B3) that are structurally and functionally related to estrogen receptors alpha and beta. Herein we test the hypothesis that ERRalpha regulates transcription of the genes encoding the enzymes involved in adrenal steroid production. Real-time RT-PCR was first used to determine the levels of ERRalpha mRNA in various human tissues. Adult adrenal levels of ERRalpha transcript were similar to that seen in heart, which is known to highly express ERRalpha. Expression of ERRalpha in the adult adrenal was then confirmed using Western blotting and immunohistochemistry. To examine the effects of ERRalpha on steroidogenic capacity we used reporter constructs with the 5'-flanking regions of steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage (CYP11A), 3beta-hydroxysteroid dehydrogenase type II (HSD3B2), 17alpha-hydroxylase/17,20-lyase (CYP17), and dehydroepiandrosterone sulfotransferase (SULT2A1). Cotransfection of these reporter constructs with wild-type ERRalpha or VP16-ERRalpha expression vectors demonstrated ERRalpha enhanced reporter activity driven by flanking DNA from CYP17 and SULT2A1. SULT2A1 promoter activity was most responsive to the ERRalpha and VP16-ERRalpha, increasing activity 2.6- and 79.5-fold, respectively. ERRalpha effects on SULT2A1 were greater than the stimulation seen in response to steroidogenic factor 1 (SF1). Transfection of serial deletions of the 5'-flanking DNA of the SULT2A1 gene and EMSA experiments indicated the presence of three functional regulatory cis-elements which shared sequence similarity to binding sites for SF1. Taken together, the expression of ERRalpha in the adrenal and its regulation of SULT2A1 suggest an important role for this orphan receptor in the regulation of adrenal steroid production. PMID- 15878969 TI - Peroxisome proliferator-activated receptor alpha (PPARalpha) potentiates, whereas PPARgamma attenuates, glucose-stimulated insulin secretion in pancreatic beta cells. AB - Fatty acids (FAs) are known to be important regulators of insulin secretion from pancreatic beta-cells. FA-coenzyme A esters have been shown to directly stimulate the secretion process, whereas long-term exposure of beta-cells to FAs compromises glucose-stimulated insulin secretion (GSIS) by mechanisms unknown to date. It has been speculated that some of these long-term effects are mediated by members of the peroxisome proliferator-activated receptor (PPAR) family via an induction of uncoupling protein-2 (UCP2). In this study we show that adenoviral coexpression of PPARalpha and retinoid X receptor alpha (RXRalpha) in INS-1E beta cells synergistically and in a dose- and ligand-dependent manner increases the expression of known PPARalpha target genes and enhances FA uptake and beta oxidation. In contrast, ectopic expression of PPARgamma/RXRalpha increases FA uptake and deposition as triacylglycerides. Although the expression of PPARalpha/RXRalpha leads to the induction of UCP2 mRNA and protein, this is not accompanied by reduced hyperpolarization of the mitochondrial membrane, indicating that under these conditions, increased UCP2 expression is insufficient for dissipation of the mitochondrial proton gradient. Importantly, whereas expression of PPARgamma/RXRalpha attenuates GSIS, the expression of PPARalpha/RXRalpha potentiates GSIS in rat islets and INS-1E cells without affecting the mitochondrial membrane potential. These results show a strong subtype specificity of the two PPAR subtypes alpha and gamma on lipid partitioning and insulin secretion when systematically compared in a beta-cell context. PMID- 15878970 TI - Integrin activates receptor-like protein tyrosine phosphatase alpha, Src, and Rho to increase prolactin gene expression through a final phosphatidylinositol 3 kinase/cytoskeletal pathway that is additive with insulin. AB - We previously showed that receptor-like protein tyrosine phosphatase (RPTP)-alpha inhibited insulin-increased prolactin gene transcription. Others suggested that RPTPalpha was a key intermediary between integrins and activation of Src. We present evidence that inhibition of insulin-increased prolactin gene transcription was secondary to RPTPalpha activation of Src, reflecting its role as mediator of integrin responses. Src kinase activity was increased in GH4 cells transiently or stably expressing RPTPalpha and cells plated on the integrin alpha5beta1 ligand fibronectin. C-terminal Src kinase inactivated Src and blocked RPTPalpha inhibition of insulin-increased prolactin gene transcription. Expression of dominant-negative Src also prevented the RPTPalpha-mediated inhibition of insulin-increased prolactin gene expression. Low levels of a constitutively active Src mutant (SrcY/F) stimulated whereas higher expression levels of Src Y/F inhibited prolactin gene expression. Src-increased prolactin gene transcription was inhibited by expression of a blocking Rho-mutant (RhoN19), suggesting that Src acted through or required active Rho. Experiments with an activated Rho-mutant (RhoL63) demonstrated a biphasic activation/repression of prolactin gene transcription that was similar to the effect of Src. The effects of both Src and Rho were phosphatidylinositol 3-kinase dependent. Expression of SrcY/F or RhoL63 altered the actin cytoskeleton and morphology of GH4 cells. Taken together, these data suggest a physiological pathway from the cell matrix to increased prolactin gene transcription mediated by RPTPalpha/Src/Rho/phosphatidylinositol 3-kinase and cytoskeletal change that is additive with effects of insulin. Over activation of this pathway, however, caused extreme alteration of the cytoskeleton that blocked activation of the prolactin gene. PMID- 15878971 TI - Strain-dependent influences on the hypothalamo-pituitary-adrenal axis profoundly affect the 7B2 and PC2 null phenotypes. AB - Two null mouse models have previously been created to study the role of the prohormone convertase (PC2) and its helper protein 7B2; unexpectedly, the phenotypes of these two nulls differ profoundly, with the 7B2 but not the PC2 null dying at 5 wk. The genetic backgrounds of these two models differ, with the 7B2 null in a 129/SvEv (129) background and the PC2 null in a mixed C57BL/N6:129/SvEv (B6:129) background. Because background can contribute greatly to phenotype, we have here examined strain influence on the hypothalamo-pituitary adrenal (HPA) axis and glucose levels in wild-type, 7B2 null, and PC2 null mice. Wild-type B6 and 129 mice differed in basal corticosterone and glucose levels. When 7B2 nulls were transferred onto the B6 background, they survived and showed greatly decreased circulating corticosterone and increased blood glucose levels, most likely due to the comparatively higher adrenal resistance of the B6 strain to ACTH stimulation. Circulating ACTH levels were increased over wild-type in the B6 7B2 null but did not reach levels as high as the 129 7B2 null. Conversely, when the mixed-strain PC2 nulls were bred into the 129 background at the N6 generation, they began to exhibit the Cushing's-like phenotype characteristic of 129 7B2 null mice and died before 6 wk of age. Taken together, these results indicate that background effects are critical because they increase the phenotypic differences between the 7B2 and PC2 nulls and play a life-or-death role in the ACTH hypersecretion syndrome present in both 129 nulls. PMID- 15878972 TI - Human short-term repopulating stem cells are efficiently detected following intrafemoral transplantation into NOD/SCID recipients depleted of CD122+ cells. AB - The nonobese diabetic/severe combined immune deficiency (NOD/SCID) xenotransplantation model has emerged as a widely used assay for human hematopoietic stem cells; however, barriers still exist that limit engraftment. We previously identified a short-term SCID-repopulating cell (SRC) following direct intrafemoral injection into NOD/SCID mice, whereas others characterized similar SRCs using NOD/SCID mice depleted of natural killer (NK) cell activity. To determine the model that most efficiently detects short-term SRCs, we compared human engraftment in 6 different xenotransplantation models: NOD/SCID-beta2 microglobulin-null mice, anti-CD122 (interleukin-2 receptor beta [IL-2Rbeta]) treated or unmanipulated NOD/SCID mice, each given transplants by intravenous or intrafemoral injection. Human cell engraftment was highest in intrafemorally injected anti-CD122-treated NOD/SCID mice compared to all other groups at 2 and 6 weeks after transplantation. These modifications to the SRC assay provide improved detection of human stem cells and demonstrate that CD122+ cells provide barriers to stem cell engraftment, a finding with potential clinical relevance. PMID- 15878973 TI - Global approach to the diagnosis of leukemia using gene expression profiling. AB - Accurate diagnosis and classification of leukemias are the bases for the appropriate management of patients. The diagnostic accuracy and efficiency of present methods may be improved by the use of microarrays for gene expression profiling. We analyzed gene expression profiles in 937 bone marrow and peripheral blood samples from 892 patients with all clinically relevant leukemia subtypes and from 45 nonleukemic controls by U133A and U133B GeneChip arrays. For each subgroup, differentially expressed genes were calculated. Class prediction was performed using support vector machines. Prediction accuracy was estimated by 10 fold cross-validation and was assessed for robustness in a 100-fold resampling approach using randomly chosen test sets consisting of one third of the samples. Applying the top 100 genes of each subgroup, an overall prediction accuracy of 95.1% was achieved that was confirmed by resampling (median, 93.8%; 95% confidence interval, 91.4%-95.8%). In particular, acute myeloid leukemia (AML) with t(15;17), AML with t(8;21), AML with inv(16), chronic lymphatic leukemia (CLL), and pro-B-cell acute lymphoblastic leukemia (pro-B-ALL) with t(11q23) were classified with 100% sensitivity and 100% specificity. Accordingly, cluster analysis completely separated all 13 subgroups analyzed. Gene expression profiling can predict all clinically relevant subentities of leukemia with high accuracy. PMID- 15878974 TI - Prospective evaluation of 2 acute graft-versus-host (GVHD) grading systems: a joint Societe Francaise de Greffe de Moelle et Therapie Cellulaire (SFGM-TC), Dana Farber Cancer Institute (DFCI), and International Bone Marrow Transplant Registry (IBMTR) prospective study. AB - The most commonly used grading system for acute graft-versus-host disease (aGVHD) was introduced 30 years ago by Glucksberg; a revised system was developed by the International Bone Marrow Transplant Registry (IBMTR) in 1997. To prospectively compare the 2 classifications and to evaluate the effect of duration and severity of aGVHD on survival, we conducted a multicenter study of 607 patients receiving T-cell-replete allografts, scored weekly for aGVHD in 18 transplantation centers. Sixty-nine percent of donors were HLA-identical siblings and 28% were unrelated donors. The conditioning regimen included total body irradiation in 442 (73%) patients. The 2 classifications performed similarly in explaining variability in survival by aGVHD grade, although the Glucksberg classification predicted early survival better. There was less physician bias or error in assigning grades with the IBMTR scoring system. With either system, only the maximum observed grade had prognostic significance for survival; neither time of onset nor progression from an initially lower grade of aGVHD was associated with survival once maximum grade was considered. Regardless of scoring system, aGVHD severity accounted for only a small percentage of observed variation in survival. Validity of these results in populations receiving peripheral blood transplants or nonmyeloablative conditioning regimens remains to be tested. PMID- 15878975 TI - Disseminated intravascular coagulation associated with acute hemoglobinemia or hemoglobinuria following Rh(0)(D) immune globulin intravenous administration for immune thrombocytopenic purpura. AB - The Food and Drug Administration (FDA) licensed Rh(o)(D) immune globulin intravenous (anti-D IGIV) on March 24, 1995, for treatment of immune thrombocytopenic purpura (ITP). A previous review described data on 15 patients who experienced acute hemoglobinemia or hemoglobinuria following anti-D IGIV administration for ITP or secondary thrombocytopenia. Eleven of those patients also experienced clinically compromising anemia, transfusion with packed red blood cells, renal insufficiency, dialysis, or death. That review suggested that patients receiving anti-D IGIV be monitored for those and other potential complications of hemoglobinemia, particularly disseminated intravascular coagulation (DIC). Through November 30, 2004, the FDA received 6 reports of DIC associated with "acute hemolysis" (or similar terms), 5 of which involved fatalities. The attending or consulting physicians assessed that acute hemolysis or DIC caused or contributed to each death. This review presents the first case series of DIC associated with acute hemoglobinemia or hemoglobinuria following anti-D IGIV administration for ITP. The purpose of this review is to increase awareness among physicians and other health care professionals that DIC may be a rare but potentially severe complication of anti-D IGIV treatment. Increased awareness of DIC as a diagnostic possibility may enable prompt recognition and medical intervention in affected patients. PMID- 15878976 TI - Bcl10 can promote survival of antigen-stimulated B lymphocytes. AB - To understand the nature of negative responses through the B-cell antigen receptor (BCR), we have screened an expression cDNA library for the ability to block BCR-induced growth arrest and apoptosis in the immature B-cell line, WEHI 231. We isolated multiple copies of full-length, unmutated Bcl10, a signaling adaptor molecule encoded by a gene found to translocate to the immunoglobulin heavy chain (IgH) locus in some mucosa-associated lymphoid tissue (MALT) lymphomas. A conditionally active form of B-cell lymphoma 10 (Bcl10) protected WEHI-231 cells from BCR-induced apoptosis upon activation. Induction of Bcl10 activity caused rapid activation of nuclear factor-kappaB (NF-kappaB) and c-Jun N terminal kinase (JNK), but not activation of extracellular signal-regulated kinase (ERK) or p38 mitogen-activated protein (MAP) kinases. These results support genetic and biochemical experiments that have implicated Bcl10 and its binding partners Carma1 and MALT1 in mediating the ability of the BCR to activate NF-kappaB. The ability of Bcl10 expression to prevent BCR-induced growth arrest and apoptosis of WEHI-231 cells was dependent on NF-kappaB activation. Finally, overexpression of Bcl10 in primary B cells activated ex vivo promoted the survival of these cells after removal of activating stimuli. Taken together these results support the hypothesis that enhanced BCL10 expression caused by translocation to the IGH locus can promote formation of MALT lymphomas. PMID- 15878977 TI - IL-3 is a potential inhibitor of osteoblast differentiation in multiple myeloma. AB - Bone destruction in multiple myeloma is characterized both by markedly increased osteoclastic bone destruction and severely impaired osteoblast activity. We reported that interleukin-3 (IL-3) levels are increased in bone marrow plasma of myeloma patients compared with healthy controls and that IL-3 stimulates osteoclast formation. However, the effects of IL-3 on osteoblasts are unknown. Therefore, to determine if IL-3 inhibits osteoblast growth and differentiation, we treated primary mouse and human marrow stromal cells with IL-3 and assessed osteoblast differentiation. IL-3 inhibited basal and bone morphogenic protein-2 (BMP-2)-stimulated osteoblast formation in a dose-dependent manner without affecting cell growth. Importantly, marrow plasma from patients with high IL-3 levels inhibited osteoblast differentiation, which could be blocked by anti-IL-3. However, IL-3 did not inhibit osteoblast differentiation of osteoblastlike cell lines. In contrast, IL-3 increased the number of CD45+ hematopoietic cells in stromal-cell cultures. Depletion of the CD45+ cells abolished the inhibitory effects of IL-3 on osteoblasts, and reconstitution of the cultures with CD45+ cells restored the capacity of IL-3 to inhibit osteoblast differentiation. These data suggest that IL-3 plays a dual role in the bone destructive process in myeloma by both stimulating osteoclasts and indirectly inhibiting osteoblast formation. PMID- 15878978 TI - The fully human anti-CD30 antibody 5F11 activates NF-{kappa}B and sensitizes lymphoma cells to bortezomib-induced apoptosis. AB - 5F11, a fully human monoclonal antibody directed against CD30, effectively induces killing of CD30-expressing lymphoma cell lines in vitro and in animal models. A recently conducted phase 1/2 study shows that 5F11 is well tolerated in heavily pretreated patients with relapsed and refractory CD30(+) lymphoma and has some clinical activity. In the present study, we demonstrate that 5F11 activates nuclear factor kappaB (NF-kappaB) and the anti-apoptotic protein cellular FLICE (Fas-associating protein with death domain-like interleukin-1beta-converting enzyme) inhibitory protein (c-flip) in Hodgkin lymphoma (HD)-derived cell lines, which might cause apoptosis resistance, thus limiting the clinical use of 5F11. To overcome this resistance, we combined 5F11 with the proteasome inhibitor bortezomib, which has been shown to suppress NF-kappaB activity. This combination revealed a synergistic cytotoxic effect in vitro and in a human HD xenograft model provided that 5F11 precedes bortezomib treatment. We conclude that initial 5F11-mediated NF-kappaB signaling sensitizes the tumor cells to bortezomib induced cell death. These data suggest a therapeutic value of this combination for HD patients. PMID- 15878979 TI - Sequential activation of class IB and class IA PI3K is important for the primed respiratory burst of human but not murine neutrophils. AB - It is well established that preexposure of human neutrophils to proinflammatory cytokines markedly augments the production of reactive oxygen species (ROS) to subsequent stimuli. This priming event is thought to be critical for localizing ROS to the vicinity of the inflammation, maximizing their role in the resolution of the inflammation, and minimizing the damage to surrounding tissue. We have used a new generation of isoform-selective phosphoinositide 3-kinase (PI3K) inhibitors to show that ROS production under these circumstances is regulated by temporal control of class I PI3K activity. Stimulation of tumor necrosis factor alpha (TNF-alpha)-primed human neutrophils with N-formyl-methionyl-leucyl phenylalanine (fMLP) results in biphasic activation of PI3K; the first phase is largely dependent on PI3Kgamma, and the second phase is largely dependent on PI3Kdelta. The second phase of PI3K activation requires the first phase; it is this second phase that is augmented by TNF-alpha priming and that regulates parallel activation of ROS production. Surprisingly, although TNF-alpha-primed mouse bone marrow-derived neutrophils exhibit superficially similar patterns of PI3K activation and ROS production in response to fMLP, these responses are substantially lower and largely dependent on PI3Kgamma alone. These results start to define which PI3K isoforms are responsible for modulating neutrophil responsiveness to infection and inflammation. PMID- 15878980 TI - Effective induction of naive and recall T-cell responses by targeting antigen to human dendritic cells via a humanized anti-DC-SIGN antibody. AB - Current dendritic cell (DC)-based vaccines are based on ex vivo-generated autologous DCs loaded with antigen prior to readministration into patients. A more direct and less laborious strategy is to target antigens to DCs in vivo via specific surface receptors. Therefore, we developed a humanized antibody, hD1V1G2/G4 (hD1), directed against the C-type lectin DC-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) to explore its capacity to serve as a target receptor for vaccination purposes. hD1 was cross-linked to a model antigen, keyhole limpet hemocyanin (KLH). We observed that the chimeric antibody-protein complex (hD1-KLH) bound specifically to DC-SIGN and was rapidly internalized and translocated to the lysosomal compartment. To determine the targeting efficiency of hD1-KLH, monocyte-derived DCs and peripheral blood lymphocytes (PBLs) were obtained from patients who had previously been vaccinated with KLH-pulsed DCs. Autologous DCs pulsed with hD1-KLH induced proliferation of patient PBLs at a 100-fold lower concentration than KLH-pulsed DCs. In addition, hD1-KLH-targeted DCs induced proliferation of naive T cells recognizing KLH epitopes in the context of major histocompatibility complex (MHC) classes I and II. We conclude that antibody-mediated targeting of antigen to DCs via DC-SIGN effectively induces antigen-specific naive as well as recall T-cell responses. This identifies DC-SIGN as a promising target molecule for DC-based vaccination strategies. PMID- 15878981 TI - Negative feedback regulation of T helper type 1 (Th1)/Th2 cytokine balance via dendritic cell and natural killer T cell interactions. AB - The ability of extracellular stimuli to modulate dendritic cell (DC) activation of natural killer T (NKT) cells was not well understood. We investigated the effects of the T helper type 1 (Th1)/Th2-cytokine environment on DC induction of NKT cell-mediated cytokine production in mice. Pretreatment of myeloid DCs with Th1 or Th2 cytokines, interleukin (IL)-4 or interferon (IFN)-gamma, led to the enhanced production of reciprocal cytokines by NKT cells (eg, IL-4 pretreatment led to the enhanced production of Th1 cytokines) in vitro and in vivo. Thus, the recognition of Th1 or Th2 cytokines by DCs acts as a negative feedback loop to maintain Th1/Th2-cytokine balance via NKT cell functions. Using these data, we manipulated cytokine levels and innate cytolytic activity in vivo to increase an antitumor response. This is the first description of a novel regulation system governing Th1/Th2 cytokine balance involving DCs and NKT cells. PMID- 15878982 TI - Rapamycin stimulates apoptosis of childhood acute lymphoblastic leukemia cells. AB - The phosphatidyl-inositol 3 kinase (PI3k)/Akt pathway has been implicated in childhood acute lymphoblastic leukemia (ALL). Because rapamycin suppresses the oncogenic processes sustained by PI3k/Akt, we investigated whether rapamycin affects blast survival. We found that rapamycin induces apoptosis of blasts in 56% of the bone marrow samples analyzed. Using the PI3k inhibitor wortmannin, we show that the PI3k/Akt pathway is involved in blast survival. Moreover, rapamycin increased doxorubicin-induced apoptosis even in nonresponder samples. Anthracyclines activate nuclear factor kappaB (NF-kappaB), and disruption of this signaling pathway increases the efficacy of apoptogenic stimuli. Rapamycin inhibited doxorubicin-induced NF-kappaB in ALL samples. Using a short interfering (si) RNA approach, we demonstrate that FKBP51, a large immunophilin inhibited by rapamycin, is essential for drug-induced NF-kappaB activation in human leukemia. Furthermore, rapamycin did not increase doxorubicin-induced apoptosis when NF kappaB was overexpressed. In conclusion, rapamycin targets 2 pathways that are crucial for cell survival and chemoresistance of malignant lymphoblasts--PI3k/Akt through the mammalian target of rapamycin and NF-kappaB through FKBP51- suggesting that the drug could be beneficial in the treatment of childhood ALL. PMID- 15878983 TI - Providing research results to study participants: support versus practice of researchers presenting at the American Society of Hematology annual meeting. AB - Offering to provide research results to study participants is gaining increasing support based, in part, on the principle of respect for persons. The frequency and means of this practice is unknown in national and international research communities. All investigators who presented oral abstracts involving human research at the American Society of Hematology Annual Meeting (December 2003) were surveyed. Responses were received from 197 (42%) of 472 eligible investigators. Nonrespondents did not differ in study type or country of origin. Only 30% (n = 48) of those who completed the survey had a formal plan for the return of research results; 40% of these would return both a summary plus individual level results. Of the respondents, 69% (n = 109) supported or strongly supported the practice; only 3% opposed the practice. The most commonly cited reasons for not returning results were: did not consider it (38%), anticipated contact difficulties (32%), and participant difficulty understanding results (26%). Only 11 (7%) indicated that their institutional review board (IRB) mandates the offer to provide results to all participants; this did not vary significantly by country. Given the high level of support in the international research community, evaluation of well-planned interventions for offering to provide research results to participants should be a priority. PMID- 15878984 TI - Toll-like receptor-4 regulation of hepatic Cyp3a11 metabolism in a mouse model of LPS-induced CNS inflammation. AB - Central nervous system (CNS) infection and inflammation severely reduce the capacity of cytochrome P-450 metabolism in the liver. We developed a mouse model to examine the effects of CNS inflammation on hepatic cytochrome P-450 metabolism. FVB, C57BL/6, and C3H/HeouJ mice were given Escherichia coli LPS (2.5 microg) by intracerebroventricular (ICV) injection. The CNS inflammatory response was confirmed by the elevation of TNF-alpha and/or IL-1beta proteins in the brain. In all mouse strains, LPS produced a 60-70% loss in hepatic Cyp3a11 expression and activity compared with saline-injected controls. Adrenalectomy did not prevent the loss in Cyp3a11 expression or activity, thereby precluding the involvement of the hypothalamic-adrenal-pituitary axis. Endotoxin was detectable (1-10 ng/ml) in serum between 15 and 120 min after ICV dosing of 2.5 microg LPS. Peripheral administration of 2.5 microg LPS by intraperitoneal injection produced similar serum endotoxin levels and a similar loss (60%) in Cyp3a11 expression and activity in the liver. The loss of Cyp3a11 in response to centrally or peripherally administered LPS could not be evoked in Toll-like receptor-4 (TLR4) mutant (C3H/HeJ) mice, indicating that TLR4 signaling pathways are directly involved in the enzyme loss. In summary, we conclude that LPS is transferred from the brain to the circulation in significant quantities in a model of CNS infection or inflammation. Subsequently, LPS that has reached the circulation stimulates a TLR4-dependent mechanism in the periphery, evoking a reduction in Cyp3a11 expression and metabolism in the liver. PMID- 15878985 TI - Glutamine inhibits cytokine-induced apoptosis in human colonic epithelial cells via the pyrimidine pathway. AB - Glutamine (Gln) prevents apoptosis in intestinal epithelial cells, but the mechanism(s) remain unknown. Gln-derived metabolites include ammonia, glutamate (Glu), glutathione (GSH), and nucleotides. We previously showed that Gln potently inhibited apoptosis in cytokine-treated human colonic HT-29 cells; this effect was specific to Gln, unaffected by Glu, and unrelated to intracellular GSH. The current research examines mechanism(s) for Gln-induced antiapoptotic effects in HT-29 cells treated with TNF-alpha-related apoptosis-inducing ligand (TRAIL). Proliferating cells were treated with Gln or selected Gln metabolites for 24 h. Cells were then treated with TRAIL and Gln or its downstream metabolites, and apoptosis was assessed at 8 h after treatment. The purine and pyrimidine precursors inosine and orotate inhibited TRAIL-induced apoptosis. However, inhibition of purine synthesis with azaserine did not alter the potent antiapoptotic effect of Gln. In contrast, the pyrimidine synthesis inhibitor, acivicin, completely prevented this response. Supplementation with the pyrimidine uracil or the pyrimidine precursor orotate rescued the acivicin-induced blockade of Gln antiapoptotic action. Removal of bicarbonate, a substrate for pyrimidine synthesis, also inhibited the antiapoptotic effects of Gln. Uracil and thymine alone also significantly decreased TRAIL-induced apoptosis. The antiapoptotic effects of Gln were independent of DNA/RNA synthesis as measured by flow cytometry and bromodeoxyuridine incorporation. In conclusion, Gln prevents TRAIL induced apoptosis in HT-29 cells through a mechanism involving the pyrimidine pathway. Our data also demonstrate the novel antiapoptotic effects of pyrimidine bases and their precursor orotate in these human intestinal cells. PMID- 15878986 TI - Light, the circadian clock, and sugar perception in the control of lignin biosynthesis. AB - Experiments were undertaken to investigate some of the mechanisms that may function to regulate lignin biosynthesis (lignification) in Arabidopsis thaliana. Northern blot analyses revealed that several genes encoding enzymes involved in the synthesis of lignin monomers displayed significant changes in transcript abundance over a diurnal cycle. Northern blot analysis also suggested that some of the changes in diurnal transcript abundance were likely to be attributable to circadian regulation, whereas others were likely to be attributable to light perception. Comparison of circadian changes in transcript abundance of lignin biosynthetic genes between wild-type plants and the sex1 mutant, which is impaired in starch turnover, suggested that carbon availability related to starch turnover might determine the capacity to synthesize lignins. This hypothesis was supported by the observation that the sex1 mutant accumulated fewer lignins than wild-type plants. Consistent with the relationship between carbon availability and lignin accumulation, analysis of dark-grown wild-type A. thaliana seedlings uncovered a role for sugars in the regulation of lignin biosynthesis. Analysis of lignin accumulation, as determined by qualitative changes in phloroglucinol staining, suggested that metabolizable sugars positively influence the abundance of lignins. Transcriptome analysis supports the hypothesis that sugars are not merely a source of carbon skeletons for lignification, but they also function as a signal to enhance the capacity to synthesize lignins. PMID- 15878987 TI - Global divergence of microbial genome sequences mediated by propagating fronts. AB - We model the competition between homologous recombination and point mutation in microbial genomes, and present evidence for two distinct phases, one uniform, the other genetically diverse. Depending on the specifics of homologous recombination, we find that global sequence divergence can be mediated by fronts propagating along the genome, whose characteristic signature on genome structure is elucidated, and apparently observed in closely related Bacillus strains. Front propagation provides an emergent, generic mechanism for microbial "speciation," and suggests a classification of microorganisms on the basis of their propensity to support propagating fronts. PMID- 15878988 TI - A late Neandertal femur from Les Rochers-de-Villeneuve, France. AB - In 2002, a Neandertal partial femoral diaphysis was discovered at Les Rochers-de Villeneuve (Vienne, France). Radiocarbon dated to approximately 40,700 14C years before present, this specimen is one of the most recent Middle Paleolithic Neandertals. The diaphysis derives from an archeological level indicating alternating human and carnivore (mostly hyena) occupation of the cave, reinforcing the close proximity and probable competition of Middle Paleolithic humans with large carnivores for resources and space. Morphological aspects of the diaphysis and ancient DNA extracted from it indicate that it is aligned with the Neandertals and is distinct from early modern humans. However, its midshaft cortical bone distribution places it between other Middle Paleolithic Neandertals and the Chatelperronian Neandertal from La Roche-a-Pierrot, supporting a pattern of changing mobility patterns among late Middle Paleolithic Neandertals on the eve of modern human dispersals into Europe. PMID- 15878989 TI - Interleukin 10 attenuates neointimal proliferation and inflammation in aortic allografts by a heme oxygenase-dependent pathway. AB - Interleukin 10 (IL-10) is a pleiotropic cytokine with well known antiinflammatory, immunosuppressive, and immunostimulatory properties. Chronic allograft rejection, characterized by vascular neointimal proliferation, is a major cause of organ transplant loss, particularly in heart and kidney transplant recipients. In a Dark Agouti to Lewis rat model of aortic transplantation, we evaluated the effects of a single intramuscular injection of a recombinant adeno associated viral vector (serotype 1) encoding IL-10 (rAAV1-IL-10) on neointimal proliferation and inflammation. rAAV1-IL-10 treatment resulted in a significant reduction of neointimal proliferation and graft infiltration with macrophages and T and B lymphocytes. The mechanism underlying the protective effects of IL-10 in aortic allografts involved heme oxygenase 1 (HO-1) because inhibition of HO activity reversed not only neointimal proliferation but also inflammatory cell infiltration. Our results indicate that IL-10 attenuates neointimal proliferation and inflammatory infiltration and strongly imply that HO-1 is an important intermediary through which IL-10 regulates the inflammatory responses associated with chronic vascular rejection. PMID- 15878990 TI - Emotion expression in human punishment behavior. AB - Evolutionary theory reveals that punishment is effective in promoting cooperation and maintaining social norms. Although it is accepted that emotions are connected to punishment decisions, there remains substantial debate over why humans use costly punishment. Here we show experimentally that constraints on emotion expression can increase the use of costly punishment. We report data from ultimatum games, where a proposer offers a division of a sum of money and a responder decides whether to accept the split, or reject and leave both players with nothing. Compared with the treatment in which expressing emotions directly to proposers is prohibited, rejection of unfair offers is significantly less frequent when responders can convey their feelings to the proposer concurrently with their decisions. These data support the view that costly punishment might itself be used to express negative emotions and suggest that future studies will benefit by recognizing that human demand for emotion expression can have significant behavioral consequences in social environments, including families, courts, companies, and markets. PMID- 15878991 TI - Structural and functional similarities between the capsid proteins of bacteriophages T4 and HK97 point to a common ancestry. AB - Gene product (gp) 24 of bacteriophage T4 forms the pentameric vertices of the capsid. Using x-ray crystallography, we found the principal domain of gp24 to have a polypeptide fold similar to that of the HK97 phage capsid protein plus an additional insertion domain. Fitting gp24 monomers into a cryo-EM density map of the mature T4 capsid suggests that the insertion domain interacts with a neighboring subunit, effecting a stabilization analogous to the covalent crosslinking in the HK97 capsid. Sequence alignment and genetic data show that the folds of gp24 and the hexamer-forming capsid protein, gp23*, are similar. Accordingly, models of gp24* pentamers, gp23* hexamers, and the whole capsid were built, based on a cryo-EM image reconstruction of the capsid. Mutations in gene 23 that affect capsid shape map to the capsomer's periphery, whereas mutations that allow gp23 to substitute for gp24 at the vertices modify the interactions between monomers within capsomers. Structural data show that capsid proteins of most tailed phages, and some eukaryotic viruses, may have evolved from a common ancestor. PMID- 15878992 TI - Ventralized dorsal telencephalic progenitors in Pax6 mutant mice generate GABA interneurons of a lateral ganglionic eminence fate. AB - The transcription factor Pax6 is expressed by progenitors in the ventricular zone (VZ) of dorsal telencephalon (dTel), which generate all cortical glutamatergic neurons, but not by progenitors in the medial ganglionic eminence (MGE), which generate cortical GABAergic interneurons (GABA INs), or the lateral ganglionic eminence (LGE), which generate GABA INs that normally migrate to the olfactory bulb. We show that perinatally, Pax6(sey/sey) mice, which lack functional Pax6 protein, have large subpial ectopias in dTel and ventral telencephalon connected by cell streams arising from an aberrant paraventricular ectopia found throughout dTel. The subpial and paraventricular ectopias and connecting cell streams are comprised of postmitotic neurons expressing markers for GABA INs characteristic of a LGE fate. Marker analyses show that dTel VZ progenitors in Pax6 mutants are progressively ventralized, acquiring expression of regulatory genes normally limited to GE progenitors; by midneurogenesis, the entire dTel VZ exhibits ventralization. This ventralization of the dTel VZ is paralleled by the expression of markers for GABA INs superficial to it, suggesting that it ectopically produces GABA INs, leading to their ectopias and a thinner cortical plate due to diminished production of glutamatergic neurons. Genetic lineage tracing demonstrates that the GABA INs comprising the ectopias are from a cortical Emx1 lineage generated in the dTel VZ, definitively showing that dTel progenitors and progeny acquire a ventral, GE, fate in Pax6 mutants. Thus, Pax6 delimits the appropriate proliferative zone for GABA INs and regulates their numbers and distributions by repressing the ventral fates of dTel progenitors and progeny. PMID- 15878993 TI - The domains of a cholesterol-dependent cytolysin undergo a major FRET-detected rearrangement during pore formation. AB - FRET measurements were used to determine the domain-specific topography of perfringolysin O, a pore-forming toxin, on a membrane surface at different stages of pore formation. The data reveal that the elongated toxin monomer binds stably to the membrane in an "end-on" orientation, with its long axis approximately perpendicular to the plane of the membrane bilayer. This orientation is largely retained even after monomer association to form an oligomeric prepore complex. The domain 3 (D3) polypeptide segments that ultimately form transmembrane beta hairpins remain far above the membrane surface in both the membrane-bound monomer and prepore oligomer. Upon pore formation, these segments enter the bilayer, whereas D1 moves to a position that is substantially closer to the membrane. Therefore, the extended D2 beta-structure that connects D1 to membrane-bound D4 appears to bend or otherwise reconfigure during the prepore-to-pore transition of the perfringolysin O oligomer. PMID- 15878994 TI - The Toll pathway is important for an antiviral response in Drosophila. AB - The innate immune response of Drosophila melanogaster is governed by a complex set of signaling pathways that trigger antimicrobial peptide (AMP) production, phagocytosis, melanization, and encapsulation. Although immune responses against both bacteria and fungi have been demonstrated in Drosophila, identification of an antiviral response has yet to be found. To investigate what responses Drosophila mounts against a viral infection, we have developed an in vivo Drosophila X virus (DXV)-based screening system that identifies altered sensitivity to viral infection by using DXV's anoxia-induced death pathology. Using this system to screen flies with mutations in genes with known or suggested immune activity, we identified the Toll pathway as a vital part of the Drosophila antiviral response. Inactivation of this pathway instigated a rapid onset of anoxia induced death in infected flies and increases in viral titers compared to those in WT flies. Although constitutive activation of the pathway resulted in similar rapid onset of anoxia sensitivity, it also resulted in decreased viral titer. Additionally, AMP genes were induced in response to viral infection similar to levels observed during Escherichia coli infection. However, enhanced expression of single AMPs did not alter resistance to viral infection or viral titer levels, suggesting that the main antiviral response is cellular rather than humoral. Our results show that the Toll pathway is required for efficient inhibition of DXV replication in Drosophila. Additionally, our results demonstrate the validity of using a genetic approach to identify genes and pathways used in viral innate immune responses in Drosophila. PMID- 15878995 TI - G protein-dependent presynaptic inhibition mediated by AMPA receptors at the calyx of Held. AB - The alpha-amino-3-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) is an ionotropic receptor mediating excitatory synaptic transmission, but it can also interact with intracellular messengers. Here we report that, at the calyx of Held in the rat auditory brainstem, activation of AMPARs induced inward currents in the nerve terminal and inhibited presynaptic Ca2+ currents (I(pCa)), thereby attenuating glutamatergic synaptic transmission. The AMPAR-mediated I(pCa) inhibition was disinhibited by a strong depolarizing pulse and occluded by the nonhydrolyzable GTP analog GTPgammaS loaded into the terminal. We conclude that functional AMPARs are expressed at the calyx of Held nerve terminal and that their activation inhibits voltage-gated Ca2+ channels by an interaction with heterotrimeric GTP-binding proteins (G proteins). Thus, at a central glutamatergic synapse, presynaptic AMPARs have a metabotropic nature and regulate transmitter release by means of G proteins. PMID- 15878996 TI - Preclinical profile of ciclesonide, a novel corticosteroid for the treatment of asthma. AB - Ciclesonide is a novel, inhaled corticosteroid under development for the treatment of asthma. Ciclesonide is activated to desisobutyryl-ciclesonide (des CIC) in the lungs to provide potent anti-inflammatory activity. The investigations herein compared the activity of ciclesonide with fluticasone in animal models to assess efficacy/potency as an airway anti-inflammatory and the comparative side effect potential to consider the therapeutic ratio of each compound. In radioligand binding assays, des-CIC and fluticasone exhibited comparable high-affinity binding to the glucocorticoid receptor, whereas ciclesonide exhibited 100-fold less binding affinity. In the Brown Norway rat model of antigen-induced airway eosinophilia and in a model of Sephadex-induced lung edema, ciclesonide and fluticasone exhibited comparable efficacy. Interestingly, following 7-day intratracheal administration, ciclesonide elicited adrenal involution with a potency that was 44-fold less than fluticasone. Furthermore, ciclesonide was 22-fold less active than fluticasone in eliciting hypoplasia of the femoral growth plate. These data support the concept that ciclesonide acts as a parent compound that, when delivered to the airways, can be transformed into the active metabolite des-CIC, resulting in local high anti inflammatory activity. Furthermore, ciclesonide possesses equivalent anti inflammatory efficacy through pulmonary activation with a significantly improved safety profile in preclinical animal models compared with fluticasone. PMID- 15878997 TI - Protein kinase C betaII peptide inhibitor exerts cardioprotective effects in rat cardiac ischemia/reperfusion injury. AB - Ischemia followed by reperfusion (I/R) in the presence of polymorphonuclear leukocytes (PMNs) results in a marked cardiac contractile dysfunction. A cell permeable protein kinase C (PKC) betaII peptide inhibitor was used to test the hypothesis that PKC betaII inhibition could attenuate PMN-induced cardiac dysfunction by suppression of superoxide production from PMNs and increase NO release from vascular endothelium. The effects of the PKC betaII peptide inhibitor were examined in isolated ischemic (20 min) and reperfused (45 min) rat hearts with PMNs. The PKC betaII inhibitor (10 microM; n = 7) significantly attenuated PMN-induced cardiac dysfunction compared with I/R hearts (n = 9) receiving PMNs alone in left ventricular developed pressure (LVDP) and the maximal rate of LVDP (+dP/dt(max)) cardiac function indices (p < 0.01). The PKC betaII inhibitor at 10 microM significantly increased endothelial NO release from a basal value of 1.85 +/- 0.18 pmol NO/mg tissue to 3.49 +/- 0.62 pmol NO/mg tissue from rat aorta. It also significantly inhibited superoxide release (i.e., absorbance) from N-formyl-L-methionyl-L-leucyl-L-phenylalanine-stimulated rat PMNs from 0.13 +/- 0.01 to 0.02 +/- 0.004 (p < 0.01) at 10 microM. Histological analysis of the left ventricle of representative rat hearts from each group showed that the PKC betaII peptide inhibitor-treated hearts experienced a marked reduction in PMN vascular adherence and infiltration into the postreperfused cardiac tissue compared with I/R + PMN hearts (p < 0.01). These results suggest that the PKC betaII peptide inhibitor attenuates PMN-induced post-I/R cardiac contractile dysfunction by increasing endothelial NO release and by inhibiting superoxide release from PMNs. PMID- 15878998 TI - Two "knockout" mouse models demonstrate that aortic vasodilatation is mediated via alpha2a-adrenoceptors located on the endothelium. AB - UK-14,304 [5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6-quinoxalinamine]-mediated vasodilator responses were studied on wire myograph-mounted mouse aorta to determine the cells involved, mechanisms of action, and subtypes of alpha(2) adrenoceptors. In the presence of induced tone, UK-14,304 produced concentration related vasodilatation that was abolished by rauwolscine, N(omega)-nitro-L arginine methyl ester (L-NAME), or endothelium removal, indicating that endothelial alpha(2)-adrenoceptors can release nitric oxide. In the alpha(2A) adrenoceptor knockout mouse and the D79N mouse, a functional knockout of the alpha(2A)-adrenoceptor, these relaxant effects of UK-14,304 were lost, indicating the involvement of the alpha(2A)-adrenoceptor. UK-14,304 could also contract aorta: a small contraction occurred at high concentrations, was enhanced by L NAME, and was absent in the alpha(1D)-adrenoceptor knockout mouse, indicating activation of the alpha(1D)-adrenoceptor. There was no evidence for a contractile alpha(2)-adrenoceptor-mediated response. A fluorescent ligand, quinazoline piperazine bodipy, antagonized the relaxant action of UK-14,304. This compound could be visualized on aortic endothelial cells, and its binding could be prevented by rauwolscine, providing direct evidence for the presence of alpha(2) adrenoceptors on the endothelium. Norepinephrine reduced tone in the alpha(1D) adrenoceptor knockout and controls, an effect blocked by rauwolscine and L-NAME but not by prazosin. This suggests that norepinephrine activates endothelial alpha(2)-adrenoceptors. In conclusion, the endothelium of mouse aorta has an alpha(2A)-adrenoceptor that responds to norepinephrine; promotes the release of nitric oxide, causing smooth muscle relaxation; and that can be directly visualized. Knockout or genetic malfunction of this receptor should increase arterial stiffness, exacerbated by raised catecholamines, and contribute to heart failure. PMID- 15878999 TI - Comparison of cannabidiol, antioxidants, and diuretics in reversing binge ethanol induced neurotoxicity. AB - Binge alcohol consumption in the rat induces substantial neurodegeneration in the hippocampus and entorhinal cortex. Oxidative stress and cytotoxic edema have both been shown to be involved in such neurotoxicity, whereas N-methyl-d-aspartate (NMDA) receptor activity has been implicated in alcohol withdrawal and excitoxic injury. Because the nonpsychoactive cannabinoid cannabidiol (CBD) was previously shown in vitro to prevent glutamate toxicity through its ability to reduce oxidative stress, we evaluated CBD as a neuroprotectant in a rat binge ethanol model. When administered concurrently with binge ethanol exposure, CBD protected against hippocampal and entorhinal cortical neurodegeneration in a dose-dependent manner. Similarly, the common antioxidants butylated hydroxytoluene and alpha tocopherol also afforded significant protection. In contrast, the NMDA receptor antagonists dizocilpine (MK-801) and memantine did not prevent cell death. Of the diuretics tested, furosemide was protective, whereas the other two anion exchanger inhibitors, L-644,711 [(R)-(+)-(5,6-dichloro2,3,9,9a-tetrahydro 3-oxo 9a-propyl-1H-fluoren-7-yl)oxy acetic acid] and bumetanide, were ineffective. In vitro comparison of these diuretics indicated that furosemide is also a potent antioxidant, whereas the nonprotective diuretics are not. The lack of efficacy of L-644,711 and bumetanide suggests that the antioxidant rather than the diuretic properties of furosemide contribute most critically to its efficacy in reversing ethanol-induced neurotoxicity in vitro, in our model. This study provides the first demonstration of CBD as an in vivo neuroprotectant and shows the efficacy of lipophilic antioxidants in preventing binge ethanol-induced brain injury. PMID- 15879000 TI - Renal clearance of endogenous hippurate correlates with expression levels of renal organic anion transporters in uremic rats. AB - Hippurate (HA) is a harmful uremic toxin that accumulates during chronic renal failure, and failure of the excretion system for uremic toxins is thought to be responsible. Recently, we reported that rat organic anion transporter 1 (rOat1) is the primary mediator of HA uptake in the kidney, and so now we have studied the pharmacokinetics and tissue distribution of HA after a single i.v. dose of HA to normal and 5/6 nephrectomized rats (5/6Nx rats). In control rats, the renal and biliary clearances of HA were 18.1 and 0.1 ml/min/kg, respectively. Plasma clearance decreased as dosage increased from 0.1 to 5 mg/kg, which suggests that renal tubular secretion is the primary route for elimination of HA. The plasma clearance of HA was significantly decreased in 5/6 Nx rats compared with normal rats. In 5/6 Nx rats, renal clearance of endogenous HA correlated more closely with clearance of p-aminohippurate than with that of creatinine. Protein expression of rOat1 and rOat3, assessed by Western blot analysis, was decreased in 5/6 Nx rats. Furthermore, in 5/6 Nx rats, the renal secretory clearance of endogenous HA correlated closely with protein expression of renal rOats. Thus, HA is primarily eliminated from the plasma via the kidney by active tubular secretion. The renal clearance of endogenous HA seems to be a useful indicator of changes in renal secretion that accompany the reduced levels of OAT protein in chronic renal failure. PMID- 15879001 TI - Cocaine up-regulates Fra-2 and sigma-1 receptor gene and protein expression in brain regions involved in addiction and reward. AB - Sigma receptors have recently been implicated in the actions of cocaine, and antagonists of these receptors prevent many acute and subchronic cocaine effects. A previous study revealed that the immediate early gene fra-2 is up-regulated after cocaine administration, and this effect is prevented by the sigma-1 receptor antagonist BD1063 [1-[2-(3,4-dichlorophenyl)ethyl]-4-methylpiperazine]. In the present study, the effects of cocaine and BD1063 on the expression of six fos and jun genes were evaluated in mouse brains using cDNA microarrays. Several of these genes were altered by cocaine, but only the alteration in fra-2 was prevented by BD1063. The time courses of fra-2 and sigma-1 receptor gene and protein expression in different brain regions were also determined. Cocaine up regulated fra-2, which was followed by a later up-regulation of sigma-1 receptors. The cocaine-induced up-regulation of fra-2 and sigma-1 receptor genes and proteins were detected in whole brain, striatum, and cortex, but not in cerebellum. All of these cocaine-induced effects were prevented by BD1063. The interaction between cocaine, fra-2, and sigma-1 receptors involves brain regions that are established components of the neural circuit for reward, suggesting that they may contribute to the enduring changes that underlie the cellular basis of drug abuse. PMID- 15879002 TI - Resveratrol-mediated activation of cAMP response element-binding protein through adenosine A3 receptor by Akt-dependent and -independent pathways. AB - A recent study documented a role of adenosine A(3)-Akt-cAMP response element binding protein (CREB) survival signaling in resveratrol preconditioning of the heart. In this study, we demonstrate that resveratrol-mediated CREB activation can also occur through an Akt-independent pathway. Isolated rat hearts were perfused for 15 min with Krebs-Henseleit bicarbonate (KHB) buffer containing resveratrol in the absence or presence of adenosine A(3) receptor blocker MRS 1191 [3-ethyl-5-benzyl-2-methyl-4-phenylethynyl-6-phenyl-1,4-(+/-) dihydropyridine-3,5-dicar-boxylate], phosphatidylinositol 3 (PI3)-kinase inhibitor LY294002 [2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride], mitogen-activated extracellular signal-regulated protein kinase inhibitor PD098059 [2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one], or a combination of LY294002 and PD098059. All hearts were subsequently subjected to 30-min ischemia followed by 2-h reperfusion. Cardioprotection was examined by determining infarct size, cardiomyocyte apoptosis, and ventricular recovery. Resveratrol phosphorylated both Akt and CREB that was blocked by MRS-1191, which also abolished cardioprotective abilities of resveratrol. LY294002 completely inhibited Akt phosphorylation but partially blocked the phosphorylation of CREB. Inhibition of PI3-kinase also partially blocked resveratrol's ability to precondition the heart. PD098059 partially blocked the phosphorylation of CREB and resveratrol-mediated cardioprotection. Preperfusing the hearts with LY294002 and PD098059 together completely abolished the phosphorylation of CREB, simultaneously inhibiting resveratrol-mediated cardioprotection. The results indicate that resveratrol preconditions the hearts through adenosine A(3) receptor signaling that triggers the phosphorylation of CREB through both Akt dependent and -independent pathways, leading to cardioprotection. PMID- 15879003 TI - Increased potency of a novel complement factor 5a receptor antagonist in a rat model of inflammatory bowel disease. AB - We have previously shown that complement factor 5a (C5a) plays a role in the pathogenesis of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats by using the selective, orally active C5a antagonist AcF-[OP(d-Cha)WR]. This study tested the efficacy and potency of a new C5a antagonist, hydrocinnamate (HC)-[OP(d-Cha)WR], which has limited intestinal lumenal metabolism, in this model of colitis. Analogs of AcF-[OP(d-Cha)WR] were examined for their susceptibility to alimentary metabolism in the rat using intestinal mucosal washings. One metabolically stable analog, HC-[OP(d-Cha)WR], was then evaluated pharmacokinetically and investigated at a range of doses (0.03-10 mg/kg/day p.o.) in the 8-day rat TNBS-colitis model, against the comparator drug AcF-[OP(d Cha)WR]. Using various amino acid substitutions, it was determined that the AcF moiety of AcF-[OP(d-Cha)WR] was responsible for the metabolic instability of the compound in intestinal mucosal washings. The analog HC-[OP(d-Cha)WR], equiactive in vitro to AcF-[OP(d-Cha)WR], was resistant to intestinal metabolism, but it displayed similar oral bioavailability to AcF-[OP(d-Cha)WR]. However, in the rat TNBS-colitis model, HC-[OP(d-Cha)WR] was effective at reducing mortality, colon edema, colon macroscopic scores, and increasing food consumption and body weights, at 10- to 30-fold lower oral doses than AcF-[OP(d-Cha)WR]. These studies suggest that resistance to intestinal metabolism by HC-[OP(d-Cha)WR] may result in increased local concentrations of the drug in the colon, thus affording efficacy with markedly lower oral doses than AcF-[OP(d-Cha)WR] against TNBS colitis. This large increase in potency and high efficacy of this compound makes it a potential candidate for clinical development against intestinal diseases such as inflammatory bowel disease. PMID- 15879004 TI - Effects of beta-phenylethylamine on dopaminergic neurons of the ventral tegmental area in the rat: a combined electrophysiological and microdialysis study. AB - The effects of systemic administration of beta-phenylethylamine (beta-PEA) and microiontophoretically applied beta-PEA on the spontaneous discharge of dopamine (DA) neurons in the ventral tegmental area (VTA) of the anesthetized rat were examined. Intravenous administration of beta-PEA (1.0, 2.5, and 5.0 mg/kg) and microiontophoretic applications of beta-PEA caused inhibitory responses in DA neurons. Systemic administration and microiontophoretic applications of beta-PEA induced dose- or current-dependent responses. The systemic beta-PEA-induced inhibitory responses were reversed by pretreatment with the DA D(2) receptor antagonists haloperidol (0.5 mg/kg i.p.) and sulpiride (10 mg/kg i.p). Pretreatment with reserpine (5 mg/kg i.p. 24 h earlier) did not completely block the systemic administration of beta-PEA (2.5 mg/kg) inhibition. A microdialysis study of freely moving rats demonstrated that the extracellular DA level increased significantly in response to local application of beta-PEA (100 muM) in the VTA via a microdialysis probe, and local application of beta-PEA-stimulated somatodendritic DA release in the VTA. The beta-PEA-induced release of DA was calcium ion-independent and was enhanced by pretreatment with pertussis toxin. These findings indicate that beta-phenylethylamine inhibits DA neuron activity via DA D(2) autoreceptors in the rat VTA and that this inhibitory effect is mediated by the somatodendritic DA release. PMID- 15879005 TI - Recognition of benztropine by the dopamine transporter (DAT) differs from that of the classical dopamine uptake inhibitors cocaine, methylphenidate, and mazindol as a function of a DAT transmembrane 1 aspartic acid residue. AB - Binding of cocaine to the dopamine transporter (DAT) protein blocks synaptic dopamine clearance, triggering the psychoactive effects associated with the drug; the discrete drug-protein interactions, however, remain poorly understood. A longstanding postulate holds that cocaine inhibits DAT-mediated dopamine transport via competition with dopamine for formation of an ionic bond with the DAT transmembrane aspartic acid residue D79. In the present study, DAT mutations of this residue were generated and assayed for translocation of radiolabeled dopamine and binding of radiolabeled DAT inhibitors under identical conditions. When feasible, dopamine uptake inhibition potency and apparent binding affinity K(i) values were determined for structurally diverse DAT inhibitors. The glutamic acid substitution mutant (D79E) displayed values indistinguishable from wild-type DAT in both assays for the charge-neutral cocaine analog 8-oxa-norcocaine, a finding not supportive of the D79 "salt bridge" ligand-docking model. In addressing whether the D79 side chain contributes to the DAT binding sites of other portions of the cocaine pharmacophore, only inhibitors with modifications of the tropane ring C-3 substituent, i.e., benztropine and its analogs, displayed a substantially altered dopamine uptake inhibition potency as a function of the D79E mutation. A single conservative amino acid substitution thus differentiated structural requirements for benztropine function relative to those for all other classical DAT inhibitors. Distinguishing the precise mechanism of action of this DAT inhibitor with relatively low abuse liability from that of cocaine may be attainable using DAT mutagenesis and other structure-function studies, opening the door to rational design of therapeutic agents for cocaine abuse. PMID- 15879006 TI - Resistance of human astrocytoma cells to apoptosis induced by mitochondria damaging agents: possible implications for anticancer therapy. AB - The success of anticancer chemotherapy is often hampered by resistance to apoptosis, which may depend on defects in intracellular cell death pathways. Characterizing the alterations of these pathways is a prerequisite for developing alternative and effective antitumoral strategies. Here, we investigated the susceptibility of a human astrocytoma cell line, ADF, to apoptotic cell death induced by mitochondria-damaging agents. Neither the anticancer agent betulinic acid nor the "mitochondriotropic" poisons 2-deoxy-d-ribose and potassium cyanide induced apoptosis of these cells, despite induction of highly significant mitochondrial depolarization, eventually resulting in necrotic death. Resistance to apoptosis was not due to presence of the multidrug resistance pump or to impaired expression of caspase-8, caspase-9, or "executioner" caspase-3. Cloning of caspase-9 revealed the presence of full-length caspase-9alpha and a short variant (caspase-9beta), which, in other tumors, acts as a dominant negative of the long isoform. All analyzed clones showed a point mutation in the prodomain region that is known to interact with mitochondria-released factors. Thus, in these human astrocytoma cells, mitochondria-damaging agents induce a regulated form of mitochondrial-dependent necrotic cell death (oncosis). Resistance to apoptosis is due to an intrinsic defect of caspase-9, leading to inhibition of enzyme activation and/or impaired interaction with proteins released from depolarized mitochondria. These results may have implications for developing strategies aimed at overcoming tumor resistance to chemotherapy. PMID- 15879007 TI - Acetaminophen and the cyclooxygenase-3 puzzle: sorting out facts, fictions, and uncertainties. AB - Cyclooxygenase (COX)-3, a novel COX splice variant, was suggested as the key to unlocking the mystery of the mechanism of action of acetaminophen. Although COX-3 might have COX activity in canines, and this activity might be inhibited by acetaminophen, its low expression level and the kinetics indicate unlikely clinical relevance. In rodents and humans, COX-3 encodes proteins with completely different amino acid sequences than COX-1 or COX-2 and without COX activity; therefore, it is improbable that COX-3 in these species plays a role in prostaglandin-mediated fever and pain. The aim of this review is to evaluate the literature that seeks to point out critical theoretical and methodological limitations of the COX-3 studies that led several investigators to scientifically questionable conclusions. PMID- 15879008 TI - Effect of tempol on renal cyclooxygenase expression and activity in experimental diabetes in the rat. AB - Renal cyclooxygenase (COX)-2 expression is increased in the streptozotocin (STZ) diabetic rat and is associated with enhanced renal prostaglandin release in response to arachidonic acid (AA). Endoperoxide-mediated vasoconstrictor responses to AA were also enhanced in the diabetic rat kidney. Because oxidative stress is increased in diabetes and has been shown to induce COX-2, we assessed its contribution to prostaglandin release by treating diabetic rats with tempol (120 mg/kg/day) for 28 days. Release of AA-stimulated prostaglandins PGE(2) and 6 ketoPGF(1alpha) from the isolated perfused kidney was used as an index of COX activity, and Western analysis was used to determine COX-2 protein expression. In untreated diabetic rats, the release of prostaglandins in response to AA was markedly enhanced; the increase in release of both 6-ketoPGF(1alpha) and PGE(2) after AA was twice that in control rats. Renal cortical COX-2 expression in diabetic rats was 3-fold that of control rats. Tempol treatment reduced the AA stimulated release of prostaglandins to levels seen in control rats; this was associated with reduced expression of COX-2 protein to levels not different from that in control rats. However, the enhanced vasoconstrictor response to AA in diabetic rats was unaffected by tempol treatment but abolished by inhibition of COX-1 with SC58560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-(trifluoromethyl)-1H pyrazole]. The addition of tempol to the perfusate of kidneys from diabetic and control rats had only a slight effect on prostaglandin release. We conclude that oxidative stress is an integral component of the mechanism involved in the induction of renal COX-2 in diabetes. PMID- 15879009 TI - Quality of pharmacies in Pakistan: a cross-sectional survey. AB - OBJECTIVE: To estimate the proportion of pharmacies meeting licensing requirements and to identify factors associated with these pharmacies in urban Rawalpindi, Pakistan. DESIGN: Cross-sectional questionnaire survey conducted during July-September 2001, of 311 pharmacies selected from a drug company list of 506. SETTING: Free-standing licensed and unlicensed pharmacies in urban Rawalpindi. STUDY PARTICIPANTS: A pharmacist or (if unavailable) the most experienced drug seller. RESULTS: The proportion of pharmacies meeting licensing requirements was 19.3% [95% C.I (confidence interval): 15.1, 24.2], with few qualified persons (22%). Only 10% had a temperature-monitoring device and 4% an alternative power supply for refrigerators (present in 76% of pharmacies). Associated with pharmacies meeting licensing requirements was the knowledge of not giving co-trimoxazole, a prescription drug, without prescription [OR (odds ratio) = 2.0; 95% CI: 1.1, 3.6], knowledge of the temperature range for vaccines (OR = 2.6; 95% CI: 1.4, 4.8), availability of vaccines (OR = 2.8; 95% CI: 2.8, 18.4), and alternative power supply for the refrigerator (OR = 6.0; 95% CI: 1.5, 23.7). The practice of selling drugs without prescription was not found to have a significant association (OR = 1.1; 95% CI: 0.5, 2.3); however, it did show a trend indicating discrepancy between knowledge and practice. CONCLUSIONS: Most drug sellers had fragmentary knowledge regarding drug dispensing and storage, and improper dispensing practices. There is a need to enforce existing legislation with training programmes directed towards drug sellers and to involve the pharmaceutical industry, which plays an important role in influencing pharmacy knowledge and practices. PMID- 15879010 TI - Appropriateness of total hip joint replacement. AB - OBJECTIVE: To evaluate the appropriateness of the use of total hip joint replacements. DESIGN: Observational study of consecutive patients with a diagnosis of hip osteoarthritis and who had undergone total hip arthroplasty over a 1-year period from seven hospitals. MAIN MEASURES: The appropriateness of the use of hip replacement was judged by explicit criteria developed by a panel of experts using RAND methodology. The length of hospital stay during the admission and complications were recorded 6 months post-operatively. Patients were also surveyed 6 months after discharge to determine whether they believed they had recovered or their satisfaction with the intervention. Appropriateness results of this study were compared with a previous study performed with the same criteria 4 years previously. RESULTS: In total, 784 patients participated in the study. Indications for surgery were considered necessary in 52.2% of cases, appropriate in 21.3%, uncertain in 21.4%, and inappropriate in 5.1%. Differences were found in the rates of appropriateness exclusively from one hospital. At 6 months after discharge, differences between centres were found for the proportion of patients that reported they had recovered from surgery (range 57.7-24.8%) and in the length of hospital stay during admission (range 10-16 days). Improvement in the appropriateness rates were found for all participant hospitals during both periods. CONCLUSIONS: We identified a low percentage of inappropriate indications and differences in some outcomes between centres. Compared with previously, there has been improvement in the use of this technique, although both periods are not methodologically comparable. PMID- 15879011 TI - Quality-based payment: six case examples. AB - INTRODUCTION: The logic of paying more for high-quality care and less for low quality resonates. Increasingly health system leaders worldwide acknowledge that payment reforms are needed to do just that, prompted no doubt by the growing body of evidence indicating that quality is not what it should be. PURPOSE: This review was undertaken to explore contexts in which quality-based payment appears feasible. The ultimate intent is to provoke thoughtful debate about whether and how quality-based payment might fit within a particular developing country's framework of policies to ensure and promote quality of care. METHODS: With guidance from key informants with first-hand knowledge of international quality based payment schemes, a purposive sample of six quality-based payment schemes was assembled. Schemes were examined to identify environmental contexts and design features. RESULTS: Examples illustrate a variety of approaches and a breadth of contexts in which quality-based payment has been implemented. Contrary to what might be expected, implementation does not appear to be constrained to private-sector purchasers, private-sector providers, hospital settings, nor to any particular type of underlying payment system. Further, quality-based payment pioneers are using a variety of incentive structures, and are tapping a rich mix of structural, process, and outcome standards to benchmark quality. CONCLUSION: Despite significant operational challenges, quality-based payment has been implemented in developing as well as developed countries, albeit not frequently in either instance. What we do not know--what the literature is nearly silent on- relates to the sustainability and ultimate impact of alternative incentive schemes. PMID- 15879012 TI - p53 gene (Gendicine) and embolisation overcame recurrent hepatocellular carcinoma. AB - Transcatheter arterial chemoembolisation (TACE) has become the standard treatment for unresectable hepatocellular carcinoma (HCC). However, this method is often unsuccessful. The p53 gene, which is present as a mutant form in many human tumours, is known to have broad spectrum antitumour effects when expressed normally. In this study, we report a 23 year old patient with recurrent HCC who was treated with the p53 gene (Gendicine) combining TACE, which resulted in a good clinical prognosis. PMID- 15879013 TI - In vivo CpG DNA/toll-like receptor 9 interaction induces regulatory properties in CD4+CD62L+ T cells which prevent intestinal inflammation in the SCID transfer model of colitis. AB - BACKGROUND AND METHODS: Cytosin-guanosin dinucleotide (CpG) motifs of bacterial DNA are known to be potent activators of innate immunity. We have shown previously that administration of CpG containing oligodeoxynucleotide (CpG-ODN) to mice before the onset of dextran sodium sulphate induced colitis ameliorated colitis and inhibited induction of proinflammatory cytokines. To investigate the possible involvement of CD4(+) T cells in the prophylactic CpG-ODN effects, we used the SCID transfer model of colitis. RESULTS: CD4(+)CD62L(+) T cells from CpG ODN treated donors did not induce significant intestinal inflammation in SCID recipients, in contrast with control cells. Additionally, cotransfer of these cells with CD4(+)CD62L(+) cells from normal mice protected recipient animals from colitis, indicating regulatory activity. Also, CD4(+)CD62L(+) cells from toll like receptor 9 deficient animals induced a significantly more severe colitis in SCID recipients than cells from wild-type littermate controls, suggesting a similar protective role of "endogenous" bacterial DNA leading to a less "aggressive" phenotype of these cells. There was no detectable difference in regulatory T cell surface markers between aggressive and attenuated cell pools but attenuated cell pools showed reduced proliferation in vitro and in vivo and produced less interferon gamma, interleukin (IL)-5, and IL-6 after anti-CD3 stimulation. CONCLUSIONS: Collectively, our data support the concept that both endogenous bacterial DNA and exogenously supplied CpG motifs of bacterial DNA induce regulatory properties in CD4(+) T cells. Therefore, bacterial DNA derived from the normal gut flora may contribute essentially to the homeostasis between effector and regulatory immune mechanisms in healthy individuals to protect them from chronic intestinal inflammation. PMID- 15879014 TI - Fordyce granules and hereditary non-polyposis colorectal cancer syndrome. AB - BACKGROUND: Germline mutations in mismatch repair (MMR) genes are found in only about half of clinically diagnosed families with hereditary non-polyposis colorectal cancer syndrome (HNPCC) (or Lynch syndrome). Early identification of gene carriers is essential to reduce cancer incidence and overall mortality. AIMS: Recent evidence indicates an increase in size and number of sebaceous glands following activation of the hedgehog pathway, a crucial signalling pathway for animal development that is aberrantly activated in several types of cancer. Here we sought to assess a possible association between Fordyce granules (FGs that is, ectopic sebaceous glands on the oral mucosa) and HNPCC. METHODS: A total of 15 members of five different genetically unrelated HNPCC kindreds (MLH1 gene mutation n = 8; undetectable MLH1 protein at immunochemistry n = 4; clinical diagnosis n = 3) and 630 genetically unrelated age and sex matched healthy controls were examined. Following examination of the oral mucosa surface, subjects were categorised as either FGs positive or FGs negative. RESULTS: Evidence of FGs was significantly associated with HNPCC (13/15 (86.7%) affected patients v 6/630 (0.95%) controls; p<0.0001), with a relative risk of 91.0 (95% confidence interval 40.05-206.76). The observed difference remained significant when carriers of germline mutations in MMR genes were considered (8/15 v 6/630; p<0.0001). The most common site for the FGs in HNPCC patients was the lower gingival and vestibular oral mucosa. CONCLUSIONS: Our findings suggest that a previously unrecognised activation of the sebaceous glands system occurs in HNPCC. The observation could be of value for attending physicians in identifying affected families and/or increase the accuracy of the currently available molecular genetics screenings. PMID- 15879015 TI - Levels of recent thymic emigrant cells decrease in children undergoing partial thymectomy during cardiac surgery. AB - The human thymus is required for establishment of a T-cell pool in fetal life, but postnatal thymectomy is not known to cause immunodeficiency. T-cell emigration from thymus (thymic recent emigrants [TRECs]) is a continuous thymic dependent process. We studied TREC levels pre- and post-partial thymectomy in children undergoing cardiac surgery. TRECs were quantitated by real-time PCR in peripheral blood lymphocytes of 24 children (0 to 12 years). TREC values were 47916 +/- 9271 pre-partial thymectomy and 33157 +/- 8479 post-partial thymectomy in 11 paired patients (P = 0.014). Interval between pre- and post-partial thymectomy was 8.8 days +/- 5.8 days. Another group of 8 children had 30384 +/- 9748 TRECs 16 days to 6 years post-partial thymectomy. There was a significant drop in TREC values post-partial thymectomy in the immediate postoperative period compared to prethymectomy TREC levels. While decreased thymic output may persist, the long-term implications were not evaluated in this patient population. PMID- 15879016 TI - Duplex microsphere-based immunoassay for detection of anti-West Nile virus and anti-St. Louis encephalitis virus immunoglobulin m antibodies. AB - West Nile (WN) virus was introduced into the United States in 1999, when the first human cases of WN fever and encephalitis appeared in New York City. From there, the virus has spread throughout North America, in some areas cocirculating with the related flavivirus St. Louis encephalitis (SLE) virus. Public health laboratories currently use an immunoglobulin M (IgM) antibody capture enzyme linked immunosorbent assay (MAC-ELISA) as a primary test for human serodiagnosis, followed by a confirmatory plaque-reduction neutralization test (PRNT). The MAC ELISAs take 2 days to perform; therefore there is a need for a more rapid test. This report describes a duplex microsphere-based immunoassay (MIA) that shortens the test processing time to about 4.5 h. The assay employs two sets of microspheres coupled to a single flavivirus group-reactive antibody, which are used to capture the WN and SLE viral antigens independently. Immunoglobulin G depleted serum is concurrently assayed for IgM antibodies to each of the viral antigens. The results are standardized and classified by using quadratic discriminant analysis so that a single result, anti-WN IgM-positive, anti-SLE IgM positive, negative, or nonspecific, can be determined. The duplex MIA results compared favorably to those of the plaque-reduction neutralization test and MAC ELISA. The assay proved to be reproducible, produced accurate classifications as to the infecting virus, and was specific. PMID- 15879017 TI - Boswellia carterii extract inhibits TH1 cytokines and promotes TH2 cytokines in vitro. AB - Traditional herbal formulas used to treat inflammatory arthritis in China and India include Boswellia carterii or Boswellia serrata. They both contain boswellic acids (BAs) which have been shown to exhibit anti-inflammatory and antiarthritic properties. This study tests the hypothesis that mixtures of BAs derived from B. carterii have immunomodulatory properties. B. carterii plant resin obtained from China was prepared as an ethanol extract, and the presence of seven BAs was confirmed by column chromatography, high-performance liquid chromatography, and UV laser desorption/ionization tandem mass spectroscopy. The extract was then tested for its ability to alter in vitro production of TH1 cytokines (interleukin-2 [IL-2] and gamma interferon) and TH2 cytokines (IL-4 and IL-10) by murine splenocytes. Delivery of the resin extract using ethanol as a solvent resulted in significant cellular toxicity not seen with the addition of ethanol alone. By contrast, delivery of the resin extract using a sesame oil solvent resulted in a dose-dependent inhibition of TH1 cytokines coupled with a dose-dependent potentiation of TH2 cytokines. These results indicate that a purified mixture of BAs from B. carterii plant resin exhibits carrier-dependent immunomodulatory properties in vitro. PMID- 15879018 TI - Immunoglobulin E reactivity of recombinant allergen Tyr p 13 from Tyrophagus putrescentiae homologous to fatty acid binding protein. AB - The storage mite, Tyrophagus putrescentiae, is one of the important causes of allergic disorders. Fifteen allergenic components were demonstrated in storage mite by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting, but only the group 2 allergen Tyr p 2 has been cloned and characterized. In this study, we attempted to identify and characterize new allergens from T. putrescentiae, which is a dominant species of storage mite in Korea. Expressed sequence tags were analyzed to identify possible storage mite allergens, and the cDNA sequence encoding a protein homologous to fatty acid binding protein, a mite group 13 allergen, was identified and named Tyr p 13. Its deduced amino acid sequence showed 61.1 to 85.3% identity with other mite group 13 allergens. The recombinant protein was expressed in Escherichia coli using a pET 28b vector system, and its allergenicity was investigated by enzyme-linked immunosorbent assay (ELISA). The recombinant allergen was detected in 5 of 78 (6.4%) T. putrescentiae-positive sera tested, and it inhibited 61.9% of immunoglobulin E binding to crude extract at an inhibitor concentration of 10 mug/ml by inhibition ELISA using serum from the patient who showed the strongest reaction by ELISA. In this study, a novel allergen was identified in T. putrescentiae. This allergen could be helpful for more-detailed characterizations of storage mite allergy. PMID- 15879019 TI - Analysis of human serum immunoglobulin G against O-acetyl-positive and O-acetyl negative serogroup W135 meningococcal capsular polysaccharide. AB - The capsular polysaccharide of Neisseria meningitidis serogroup W135 is expressed in both O-acetyl-positive (OA+) and O-acetyl-negative (OA-) forms. This study investigates the impact of OA status (OA+ versus OA-) on serological measurements of anti-W135 immunoglobulin G (IgG) antibodies in immunized adults. W135-specific serum antibody assignments were made for 28 postimmunization sera from adults by enzyme-linked immunosorbent assay using the meningococcal standard reference serum CDC1992. The established IgG concentration in micrograms per milliliter ([IgG]microg/ml) for CDC1992 against OA+ antigen (16.2 microg/ml) was used as a reference to assign a concentration of 10.13 microg/ml IgG against OA- antigen by cross-standardization. Overall, the IgG assignments for these sera were higher against OA+ antigen (geometric mean concentration [GMC] = 7.16 microg/ml) than against OA- antigen (GMC = 2.84 microg/ml). However, seven sera showed higher specific [IgG]microg/ml values against the OA+ antigen than against the OA- antigen. These sera were also distinguished by the inability of fluid-phase OA- antigen to compete for antibody binding to OA+ solid-phase antigen. Although there was no overall difference in functional activity measured by complement mediated serum bactericidal assay (SBA) against OA+ and OA- target bacteria (geometric mean titers of 9,642 and 9,045, respectively), three serum specimens showed a large difference in SBA antibody titers against OA+ versus OA- W135 target bacteria, which may reflect different epitope specificities for these sera. Our data indicate that, for some sera, the agreement in anti-OA+ versus anti-OA- W135 IgG assignments is serum specific and does not reflect the functional (killing) activity in vitro. PMID- 15879020 TI - Comparison of a new immunochromatographic test to enzyme-linked immunosorbent assay for rapid detection of immunoglobulin m antibodies to hepatitis e virus in human sera. AB - An immunochromatographic test for rapid detection of IgM antibodies in patients with acute hepatitis E infection was developed utilizing the well-characterized recombinant protein EP2.1 and monoclonal antibody 4B2. The new rapid test based on a novel reverse-flow technology was able to generate a positive result within 2 to 3 min. Our study showed that this test was able to detect anti-HEV IgM antibodies in 96.7% of the patient samples tested (n = 151) while maintaining an excellent specificity of 98.6% with samples from various patient or healthy control groups (total n = 208). Furthermore, this rapid test gave a good specificity of 90.9% when tested with rheumatoid factor (RF)-positive sera (RF value of < or =850 IU/ml; n = 11) although a higher concentration of RF in samples might cause cross-reactivity. The new test has a good agreement of 97.2% with a kappa value of 0.943 when compared with a reference enzyme-linked immunosorbent assay. The positive predictive value and the negative predictive value for the rapid test thus reached 98.0 and 97.6%, respectively. This is the first rapid, point-of-care test for hepatitis E and will be especially useful for the diagnosis of acute hepatitis E virus infection in field and emergency settings and in resource-poor countries. PMID- 15879021 TI - Combined administration of meningococcal serogroup B outer membrane vesicle vaccine and conjugated serogroup C vaccine indicated for prevention of meningococcal disease is safe and immunogenic. AB - MenBvac and Menjugate are safe and efficacious vaccines. The purpose of this study was to evaluate safety and immunogenicity of the combination (MenB/C) of the lyophilized active components of the conjugated group C vaccine Menjugate when reconstituted with the full liquid group B outer membrane vesicle vaccine MenBvac compared to MenBvac and Menjugate given separately. At 6-week intervals, healthy adults were given one dose of MenB/C followed by two doses of MenBvac (MenB/C group), three doses of MenBvac (MenB group), or one dose of Menjugate and two doses of placebo (MenC group). Injection site reactions were frequent in all groups. However, most reactions were short lasting and mild or moderate in intensity, and the vaccines were found to be well tolerated, with no vaccine related serious adverse events. MenB/C was immunogenic with regard to both serogroup B and C meningococci. Both the serum bactericidal assay and the enzyme linked immunosorbent assay analyses showed that the immune responses of the combination vaccine were similar to the immune responses of its separate components MenBvac and Menjugate for both serogroup B and C. In conclusion, the combined MenB/C vaccine is safe and immunogenic. The two vaccines do not interact negatively with each other and can easily be administered in the same syringe. The induced immune responses suggest that the combined vaccine is likely to confer protection against systemic group B disease caused by the vaccine strain as well as against group C meningococcal disease. PMID- 15879022 TI - Deoxycytidyl-deoxyguanosine oligonucleotide classes A, B, and C induce distinct cytokine gene expression patterns in rhesus monkey peripheral blood mononuclear cells and distinct alpha interferon responses in TLR9-expressing rhesus monkey plasmacytoid dendritic cells. AB - To determine if deoxycytidyl-deoxyguanosine oligonucleotides (CpG ODN) can be used effectively as nonspecific inducers of innate immune defenses for preventative or therapeutic interventions in infectious disease models for nonhuman primates, the present study evaluated the response of rhesus monkey peripheral blood mononuclear cells to three different synthetic CpG ODN classes by defining the cytokine gene expression patterns and by characterizing IFN alpha/beta responses. Depending on the type and dose of CpG ODN used for stimulation, distinct gene expression patterns were induced. CpG ODN class A (CpG A ODN) and CpG-C ODN, but not CpG-B ODN, were potent inducers of alpha interferon (IFN-alpha), and this response was due to IFN-alpha production by TLR9-positive plasmacytoid dendritic cells. Importantly, there was a dose-dependent increase in IFN-alpha responses to CpG-A ODN but a dose-dependent decrease in IFN-alpha responses by CpG-B ODN. The most sustained IFN-alpha response was induced by CpG A ODN and was associated with a stronger induction of interferon regulatory factor 7 and the induction of several interferon-stimulated genes. In contrast, and independent of the dose, CpG-B ODN were the weakest inducers of IFN-alpha but the most potent inducers of proinflammatory cytokines. CpG-C ODN induced cytokine gene expression patterns that were intermediate between those of CpG-A and CpG-B ODN. Thus, the different types of CpG ODN induce different post-TLR9 signaling pathways that result in distinct cytokine gene expression patterns. Based on these findings, A and C class CpG ODN, but not B class CpG ODN, may be particularly suited for use as therapeutic or prophylactic antiviral interventions. PMID- 15879023 TI - Association of selected phenotypic markers of lymphocyte activation and differentiation with perinatal human immunodeficiency virus transmission and infant infection. AB - This study of a subset of women and infants participating in National Institutes of Health Pediatric AIDS Clinical Trials Group protocol 185 evaluated lymphocyte phenotypic markers of immune activation and differentiation to determine their association with the likelihood of human immunodeficiency virus (HIV) transmission from the women to their infants and the potential for early identification and/or prognosis of infection in the infants. Lymphocytes from 215 human immunodeficiency virus type 1 (HIV)-infected women and 192 of their infants were analyzed by flow cytometry with an extended three-color panel of monoclonal antibodies. Women who did not transmit to their infants tended to have higher CD4+ T cells. Most notably, levels of total CD8+ T cells and CD8+ CD38+ cells made significant independent contributions to predicting the risk of mother-to child transmission. Adjusting for HIV-1 RNA level at entry, a one percentage point increase in these marker combinations was associated with a nine percent increase in the likelihood of maternal transmission. Total as well as naive CD4+ T cells were significantly higher in uninfected than infected infants. Total CD8+ cells, as well as CD8+ cells positive for HLA-DR+, CD45 RA+ HLA-DR+, and CD28+ HLA-DR+ were elevated in infected infants. Detailed immunophenotyping may be helpful in predicting which pregnant HIV-infected women are at increased risk of transmitting HIV to their infants. Increasing differences in lymphocyte subsets between infected and uninfected infants became apparent as early as six weeks of age. Detailed immunophenotyping may be useful in supporting the diagnosis of HIV infection in infants with perinatal HIV exposure. PMID- 15879024 TI - Association of uterine and salpingeal fibrosis with chlamydial hsp60 and hsp10 antigen-specific antibodies in Chlamydia-infected koalas. AB - Infection by Chlamydia pneumoniae or Chlamydia pecorum commonly causes chronic, fibrotic disease of the urogenital tracts of female koalas. Studies of humans have associated titers of serum immunoglobulin G (IgG) against chlamydial hsp60 and hsp10 antigens with chronic infection, salpingeal fibrosis, and tubal infertility. To determine whether a similar relationship exists in Chlamydia infected koalas, samples were collected opportunistically from 34 wild female koalas and examined by gross pathology and histopathology, PCR, and immunohistochemistry for Chlamydia spp. and enzyme-linked immunosorbent assay for serological responses to chlamydial hsp10 and hsp60 antigens. Greater anti-hsp titers occurred in Chlamydia-infected koalas with fibrous occlusion of the uterus or uterine tube than in other Chlamydia-infected koalas (for hsp10 IgG, P = 0.005; for hsp60 IgG, P = 0.001; for hsp10 IgA, P = 0.04; for hsp60 IgA, P = 0.09). However, as in humans, some koalas with tubal occlusion had low titers. Among Chlamydia-infected koalas with tubal occlusion, those with low titers were more likely to have an active component to their ongoing uterine or salpingeal inflammation (P = 0.007), such that the assay predicted, with 79% sensitivity and 92% specificity, tubal occlusion where an active component of inflammation was absent. Findings of this study permit advancement of clinical and epidemiological studies of host-pathogen-environment interactions and pose intriguing questions regarding the significance of the Th1/Th2 paradigm and antigen-presenting and inflammation-regulating capabilities of uterine epithelial cells and the roles of latency and reactivation of chlamydial infections in pathogenesis of upper reproductive tract disease of koalas. PMID- 15879025 TI - Reliability of CD4 quantitation in human immunodeficiency virus-positive children: implications for definition of immunologic response to highly active antiretroviral therapy. AB - Our objective was to develop data-based algorithms for definition of immunologic response to AIDS therapies in pediatric patients, taking account of T-cell subset measurement errors. The study design involved cross-protocol analysis of 2,148 enrollees in six completed Pediatric AIDS Clinical Trials Group trials. We used standard quantitation of T-cell subsets; linear modeling with mean-dependent measurement error variance was used to develop 95% tolerance limits for change in CD4%. For individuals with a CD4% of approximately 25%, the measurement error based 95% tolerance interval ranges from 15% to 35%, whereas for individuals with a CD4% of approximately 5%, the tolerance interval ranges from 3% to 7%. When pairs of CD4% measures taken within a time interval of less than 30 days are averaged to estimate steady-state CD4%, tolerance interval width decreases by approximately 30%. A simple graphical tool that provides a data-based criterion for immunologic response over and above variation ascribable to T-cell measurement error is provided. Variability in CD4% due to measurement error is substantial, increases with level of CD4%, and complicates assessment of immunologic response to therapy. Replicates of CD4% measures could be used to improve precision of interpretation of CD4% measures. PMID- 15879026 TI - Cytokine gene expression in response to SnSAG1 in horses with equine protozoal myeloencephalitis. AB - Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome seen in horses from the Americas and is mainly caused by Sarcocystis neurona. Recently, a 29-kDa surface antigen from S. neurona merozoites was identified as being highly immunodominant on a Western blot. This antigen has been sequenced and cloned, and the expressed protein has been named SnSAG1. In a previous study, cell-mediated immune responses to SnSAG1 were shown to be statistically significantly reduced in horses with EPM in comparison to EPM-negative control horses. It therefore appears as though the parasite is able to induce immunosuppression towards parasite-derived antigens as parasite-specific responses are decreased. Isolated peripheral blood lymphocytes from 21 EPM (cerebrospinal fluid [CSF] Western blot) negative horses with no clinical signs and 21 horses with clinical signs of EPM (CSF Western blot positive) were cocultured with SnSAG1 for 48 and 72 h, and the effect on cytokine production was investigated by means of reverse transcriptase PCR. Cytokines assayed include gamma interferon (IFN-gamma), tumor necrosis factor alpha, interleukin (IL)-2, IL-4, and IL-6. beta-Actin was used as the housekeeping gene. A Wilcoxon signed-rank test of the findings indicated that there was a statistically significant decrease in IFN-gamma production after 48 h in culture for samples from horses with clinical disease. There was also a statistically significant increase in IL-4 production after 72 h in culture for samples from horses with EPM. These results further support the notion that this parasite is able to subvert the immune system in horses with clinical disease. PMID- 15879027 TI - Development of recombinant chimeric antigen expressing immunodominant B epitopes of Leishmania infantum for serodiagnosis of visceral leishmaniasis. AB - Wild canids and domestic dogs are the main reservoir of zoonotic visceral leishmaniasis (VL) caused by Leishmania infantum (syn.: Leishmania chagasi). Serological diagnosis of VL is therefore important in both human and dog leishmaniasis from a clinical and epidemiological point of view. Routine diagnosis of VL is traditionally carried out by immunofluorescent antibody test (IFAT), which is laborious and difficult to standardize and to interpret. In the last decade, however, several specific antigens of Leishmania infantum have been characterized, allowing the development of a recombinant-based immunoassay. Among them, the whole open reading frame encoding K9 antigen, the gene fragment encoding the repetitive sequence of K26, and the 3'-terminal gene fragment of the kinesin-related protein (K39sub) were previously evaluated as diagnostic markers for canine leishmaniasis and proved to be independent in their antibody reactivity. Since sensitivity of serological test is usually higher in multiple epitope format, in this study the relevant epitopes of K9, K26, and K39 antigens were joined by PCR strategy to produce the chimeric recombinant protein. The resulting mosaic antigen was found highly expressed in Escherichia coli and efficiently purified by affinity chromatography. Antigenic properties of this recombinant antigen were evaluated by indirect enzyme-linked immunosorbent assay (ELISA) using a panel of human and dog sera previously characterized by parasitological and/or serological techniques. Chimeric ELISA showed 99% specificity in both human (n = 180) and canine (n = 343) control groups, while sensitivity was higher in canine VL (96%, n = 213) than in human VL (82%, n = 185). Accordingly, concordance between IFAT and canine chimeric ELISA (k = 0.95, 95% confidence interval = 0.93 to 0.98) was higher than between IFAT and human chimeric ELISA (k = 0.81, 95% confidence interval = 0.76 to 0.87). Results suggest the potential use of this new antigen for routine serodiagnosis of VL in both human and canine hosts. PMID- 15879028 TI - Prevalence and persistence of Chlamydia pneumoniae antibodies in healthy laboratory personnel in Finland. AB - The rates of Chlamydia pneumoniae seroconversions suggesting acute primary infections or reinfections and the prevalences of antibodies were followed up among healthy laboratory workers. Annual serum samples were collected from 47 persons in Helsinki from 1958 to 1990 and from 40 persons in Oulu from 1994 to 1999. C. pneumoniae species-specific immunoglobulin G (IgG), IgA, and IgM antibodies were measured by microimmunofluorescence (MIF) in 407 sera from Helsinki. The 185 sera collected in Oulu were tested both by MIF and by commercial enzyme immunoassay (EIA). During the follow-up periods of 31 years in Helsinki and 6 years in Oulu, seroconversions were demonstrated by MIF in 45% and 15% of the study groups, respectively. In Helsinki 9% of the persons seroconverted twice during the follow-up period. By MIF, the total incidence rate per 100 person-years at risk was 6.9 in Helsinki and 4.9 in Oulu, and annual incidence rates varied from 0 to 15.4. By EIA, annual incidence rates in Oulu varied from 0 to 10.8. The seroconversions by MIF were usually not confirmed by EIA and vice versa. Prevalence and persistence rates, respectively, of IgA antibodies were higher in EIA (62% and 26%) than in MIF (26% and 17%), whereas the figures for IgG were quite similar. The prevalence of IgG and IgA antibodies was higher in older persons than in younger ones. The presence of antibodies did not offer protection from reinfection. PMID- 15879029 TI - Enzyme-linked immunosorbent assay for pharmacological studies targeting hypoxia inducible factor 1alpha. AB - Hypoxia-inducible factor 1 (HIF-1) activates the transcription of a wide range of genes related to oxygen delivery and metabolic adaptation under hypoxic (low oxygen) conditions. HIF-1 is, in fact, a heterodimer of two subunits, HIF-1alpha and HIF-1beta. The only analytical methods available for measuring HIF-1alpha levels in tumors are immunohistochemistry and Western blotting. Immunohistochemistry has the advantage of allowing the identification and direct examination of HIF-1alpha-expressing cells, but has the intrinsic limitation, as for Western blotting, of being nonquantitative. We developed and validated an enzyme-linked immunosorbent assay (ELISA) approach to measure HIF-1alpha levels in cultured tumor cell lines in vitro. HIF-1alpha was expressed in thirteen tumor cell lines grown under hypoxic conditions; however, the levels differed strongly between cell lines. These data point to intrinsic differences between cell lines for the induction of HIF-1alpha under hypoxic conditions. The ELISA developed in the present study is thus an interesting alternative to other analytical methods used to measure HIF-1alpha protein levels and should be useful in preclinical pharmacological studies targeting HIF-1alpha. PMID- 15879030 TI - Persistence of antibodies to West Nile virus in naturally infected rock pigeons (Columba livia). AB - Wild caught rock pigeons (Columba livia) with antibodies to West Nile virus were monitored for 15 months to determine antibody persistence and compare results of three serologic techniques. Antibodies persisted for the entire study as detected by epitope-blocking enzyme-linked immunosorbent assay and plaque reduction neutralization test. Maternal antibodies in squabs derived from seropositive birds persisted for an average of 27 days. PMID- 15879031 TI - Evaluation of commercial assay detecting specific immunoglobulin g in oral fluid for determining measles immunity in vaccinees. AB - A commercial assay for detection of measles immunoglobulin G (IgG) in oral fluid was evaluated in a highly vaccinated cohort using serum IgG as gold standard. In contrast to previous studies from cohorts protected by natural immunity, antibody prevalence was significantly underestimated (-7.4%; confidence interval: -1.5 to 13.2%; P = 0.01) due to a reduced sensitivity when antibody levels were low. PMID- 15879032 TI - Detection by radioligand assay of antibodies against Borna disease virus in patients with various psychiatric disorders. AB - Using a radioligand assay, which preserves the natural form of the antigen, antibodies against Borna disease virus nucleoprotein and phosphoprotein were detected in 11 and 19 sera of 171 psychiatric patients, respectively. Compared with results by Western blotting, three and nine sera were concordantly positive, respectively. The four sera showing the highest levels of antibodies by radioligand assay were all negative by Western blotting; however, dilution and inhibition tests supported the positive results. Our results suggest the importance of conformational structure to detect human anti-Borna disease virus antibodies. PMID- 15879033 TI - Multiple autoantigen mimotopes of infectious agents induce autoimmune arthritis and uveitis in lewis rats. AB - We found multimolecular antigenic mimicry of arthritogenic autoantigens and peptides from several other "self" or foreign antigens sharing amino acid sequence homologies. Many of these new mimotopes induced arthritis and/or uveitis upon immunization in Lewis rats, indicating a role for multiple antigens in the initiation of a certain autoimmune disease. PMID- 15879034 TI - Strongyloides hyperinfection and hypogammaglobulinemia. AB - We report strongyloides hyperinfection in two patients with generalized hypogammaglobulinemia from multiple myeloma and nephrotic syndrome, despite a significant strongyloides-specific immunoglobulin G (IgG) response. In contrast to reports on animals, where human IgG was shown to be a protective antibody, our observation suggests that in humans, immunity to the infective-stage larvae is not protective against the autoinfective larvae, which are the causative agents of strongyloides hyperinfection. PMID- 15879035 TI - Construction and expression of recombinant streptolysin-o and preevaluation of its use in immunoassays. AB - Commercially available immunoassays for assessment of anti-streptolysin-O antibodies use native streptolysin-O obtained by a complex process. We prepared a biologically active recombinant streptolysin-O with higher yield and a simpler purification process. An enzyme-linked immunosorbent assay developed with this recombinant showed good correlation with a commercial test, suggesting that it could be suitable for immunoassays. PMID- 15879036 TI - As easy as ABC? Primary prevention of sexually transmitted infections. PMID- 15879037 TI - The drama of being a doctor. PMID- 15879038 TI - Management of epilepsy in women. AB - There are many aspects to the management of epilepsy in women related to their role in reproduction. Some of these need to be considered in adolescents, some are related to pregnancy, concerning both the mother and her infant, and others with the menstrual cycle and the menopause. This review considers contraception, fertility, teratogenicity, and the use of folic acid. It also discusses the special investigations in pregnancy, hyperemesis, the effect of pregnancy on the control of epilepsy, the effect of seizures on the fetus, a first fit in pregnancy, pseudoseizures, seizures during delivery, vitamin K, breast feeding, postpartum maternal epilepsy, hereditary risks, counselling, catamenial epilepsy, the menopause, and bone density. PMID- 15879039 TI - Device based treatment of heart failure. AB - As the population ages and survival from ischaemic heart disease improves, the incidence and prevalence of congestive cardiac failure has increased dramatically. Medical treatments including ACE inhibitors, beta blockers, and aldosterone antagonists have improved the outlook for most patients. However, despite optimal medical treatment there is a significant group of patients who continue to suffer poor morbidity and mortality. Device based treatment consisting of implantable cardioverter defibrillators (ICD) and cardiac resynchronisation therapy (CRT) devices offer new modes of treatment to patients with symptomatic heart failure despite optimal medical therapy. ICDs have been shown to reduce mortality in patients with severe heart failure while CRT leads to an improvement in functional class, quality of life scores, physiological measures such as peak Vo(2), and reduce hospitalisations. Combination devices, which provide both ICD and CRT functions, have now been seen to provide synergistic benefits in selected patients. PMID- 15879040 TI - Congenital heart disease in pregnancy. AB - Congenital heart disease is now more prevalent than acquired in pregnancy in the developed world. In pregnancy the fall in systemic vascular resistance and increase in blood volume and cardiac output can cause functional deterioration in certain conditions. A minimally symptomatic woman with good ventricular function, normal oxygen saturation, and no left heart obstruction should tolerate pregnancy well. Women with pulmonary hypertension or dilated aortic root (pre-replacement) should be counselled against pregnancy and given appropriate contraceptive advice. The optimum management of a pregnant woman with a metallic prosthetic valve remains to be determined. PMID- 15879042 TI - Primary prevention of sexually transmitted disease: applying the ABC strategy. AB - Escalating rates of sexually transmitted disease (STD) in many areas of the world necessitate a re-evaluation of current public health STD preventive programmes. Pervasive long term sequelae for many STD afflicted people and the emerging threat, caused by the HIV/AIDS pandemic, to some national infrastructures, suggests that ongoing initiatives focusing primarily on risk reduction through barrier protection have not met their desired objective. Recent strategies to promote non-coital sexual involvement as a means of achieving STD reduction fail to address the transmission of infection that may occur through alternative non intercourse sexual activities. The demonstrated success of the innovative, comprehensive ABC strategy shows that while risk reduction and treatment of existing infection remain important, the promotion of optimal health may be achieved more effectively through broad based comprehensive and adaptable programmes that include an emphasis on risk avoidance through delayed sexual debut and partner reduction. PMID- 15879043 TI - Multiple sclerosis: diagnosis and the management of acute relapses. AB - Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system that may result in a wide range of neurological symptoms and accumulating disability. Its course is unpredictable resulting in a changing pattern of clinical need. Diagnostic criteria for multiple sclerosis require objective evidence for dissemination in space and time. The diagnostic and management process should follow good practice guidelines with the person at the centre of the process. Appropriate support and information should be available from the time of diagnosis. Continuing education is key in enabling the person to actively participate in their management. In the event of an acute relapse the person should have direct access to the most appropriate local service. Provided medical causes have been excluded, corticosteroid treatment to hasten the recovery from the relapse should be considered. Management of an acute relapse should be comprehensive addressing any medical, functional, or psychosocial sequelae. PMID- 15879044 TI - Epistaxis: an update on current management. AB - Epistaxis is one of the commonest ENT emergencies. Although most patients can be treated within an accident and emergency setting, some are complex and may require specialist intervention. There are multiple risk factors for the development of epistaxis and it can affect any age group, but it is the elderly population with their associated morbidity who often require more intensive treatment and subsequent admission. Treatment strategies have been broadly similar for decades. However, with the evolution of endoscopic technology, new ways of actively managing epistaxis are now available. Recent evidence suggests that this, combined with the use of stepwise management plans, should limit patient complications and the need for admission. This review discusses the various treatment options and integrates the traditional methods with modern techniques. PMID- 15879045 TI - What caused the Black Death? AB - For the whole of the 20th century it was believed that the Black Death and all the plagues of Europe (1347-1670) were epidemics of bubonic plague. This review presents evidence that this view is incorrect and that the disease was a viral haemorrhagic fever, characterised by a long incubation period of 32 days, which allowed it to be spread widely even with the limited transport of the Middle Ages. It is suggested that haemorrhagic plague emerged from its animal host in Ethiopia and struck repeatedly at European/Asian civilisations, before appearing as the Black Death. The CCR5-Delta32 mutation confers protection against HIV-1 in an average of 10% of the people of European origin today. It is suggested that all the Deltaccr5 alleles originated from a single mutation event that occurred before 1000 BC and the subsequent epidemics of haemorrhagic plague gently forced up its frequency to 5 x 10(-5) at the time of the Black Death. Epidemics of haemorrhagic plague over the next three centuries then steadily raised the frequency in Europe (but not elsewhere) to present day values. PMID- 15879046 TI - Management of chronic heart failure: perceived needs of general practitioners in light of the new general medical services contract. AB - BACKGROUND: Despite the existence of several chronic heart failure (CHF) guidelines the treatment of patients with CHF is suboptimal. The new general medical services (GMS) contract in primary care has only three specific performance indicators for patients with left ventricular dysfunction. The aim of this current questionnaire survey was to assess the views of general practitioners (GPs) on CHF treatments and services in light of the new GMS contract. METHODS AND RESULTS: All local GPs (717) were sent a questionnaire. Fifty three per cent were returned. Forty five per cent of GPs had access to a community CHF nurse. Having read a national guideline (SIGN) and having the support of a CHF nurse did not seem to affect the knowledge of GPs in terms of perceived benefits of drug treatments. GPs with access to a specialist CHF nurse service attached more importance to it than those with no specialist nurse (p = 0.003). CONCLUSIONS: Most GPs were aware of the existence of a national guideline but many had not read it. There was little or no difference in the knowledge level for various evidence based treatments between GPs who had or had not read the guideline suggesting that reading guidelines may not be a key factor in determining knowledge. Support for a specialist CHF nurse was higher among GPs who already had this service, suggesting that this service is valued. The new GMS contract may improve identification and diagnosis of patients with CHF but there is a danger that it may fall short of ensuring optimal treatment for patients with CHF. PMID- 15879047 TI - Do consultants differ? Inferences drawn from hospital in-patient enquiry (HIPE) discharge coding at an Irish teaching hospital. AB - OBJECTIVE: To find out if there was a difference between hospital consultants, all trained in acute general medicine, in length of stay (LOS), re-admission rates, resource utilisation, and diagnostic coding, among patients admitted as emergencies to St James' Hospital (SJH) Dublin. METHODS: A retrospective analysis was performed of data on discharges from hospital, recorded in the hospital in patient enquiry (HIPE) system, relating to 9204 episodes among 6968 emergency medical patients admitted to SJH between 1 January 2002 and 31 October 2003. For comparative analysis, four physician groups were defined consisting of gastroenterology (GI, n = 4), respiratory (n = 3), general internal medicine (GIM, n = 2), or specialty (n = 5). RESULTS: GIM consultants had the shortest LOS (median 5 days); GIM and respiratory consultants were less likely to have long stay patients (> 30 days, p<0.0001). Patients re-admitted under the same consultant had a longer LOS than those re-admitted under a different consultant (p<0.0001). Endoscopy and GI radiology investigations were used most by GI consultants, computed tomography of the thorax by respiratory, ECHO by respiratory and specialty, and computed tomography of brain by GIM and specialty consultants. GI diagnostic codings were more frequent with GI consultants (p<0.0001), respiratory diagnoses and malignancy with respiratory (p<0.0001 for both), diabetes and hypertension with specialty (p = 0.0017), and heart failure more with GIM consultants (p = 0.001). CONCLUSIONS: This study found that the HIPE database was very powerful in predicting differences between hospital consultants in LOS, re-admission rates, resource utilisation, and disease coding. It would be of interest to examine the extent to which protocols and guidelines could reduce such variations. PMID- 15879048 TI - Starry sky appearance. PMID- 15879049 TI - Expression of granulocyte colony stimulating factor receptor in human colorectal cancer. AB - AIMS: To discover if human colorectal cancer expresses granulocyte colony stimulating factor receptor (G-CSFR) and if granulocyte colony stimulating factor (G-CSF) plays an important part in the development and progression of human colorectal cancer. METHODS: Forty two specimens of colorectal cancer and normal colorectal mucosa were investigated, taken from the colon or rectum in group of colorectal cancer patients. Immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) technique were used to show G-CSFR expression. The relation between expression of G-CSFR and clinical or pathological factors was analysed. RESULTS: Immucohistochemical analyses showed that G-CSFR was expressed in the human colorectal cancer (25 of 42, 59.52%) and seemed to be up regulated compared with the normal mucosa (14 of 42, 33.33%, p<0.001). In pronounced contrast with mostly strongly positive tumours, corresponding normal colorectal mucosa was negative or weakly positive. A significant correlation was found between G-CSFR expression and tumour stage (p = 0.001), tumour differentiation (p<0.001), but there was no significant relation between the expression of G-CSFR and the age, sex, and tumour size (p = 0.346, p = 0.686, p = 0.459). In RT-PCR, 21 of all 42 tumours had G-CSFR mRNA expression, while only 11 of 42 normal colorectal mucosa had such expression. CONCLUSIONS: These data show that G-CSFR is commonly expressed in human colorectal cancers, thus supporting a possible role for G-CSF in colorectal cancer physiology. PMID- 15879050 TI - Colonic ulcers in propylthiouracil induced vasculitis with secondary antiphospholipid syndrome. AB - A 48 year old white woman was admitted to the hospital because of several bouts of migratory polyarthritis, weight loss, fever, and abdominal pain over a period of 15 months. She had been taking propylthiouracil 100 mg daily for three years for hyperthyroidism treatment. A test for antineutrophil cytoplasmic autoantibodies (ANCA) was positive with a perinuclear pattern of staining. Antiphospholipid antibodies were also detected. Colonoscopy showed several ulcers on intestinal mucosa and the biopsy specimen showed intense microscopic vasculitis. The patient is well after methylprednisolone pulse therapy and eight months of oral azathioprine. A surveillance colonoscopy showed complete healing of intestinal ulcers. No recurrence of symptoms has occurred and autoantibodies are negative, 10 months after treatment finished. The sequence of events suggests a propylthiouracil induced vasculitis p-ANCA positive and an antiphospholipid syndrome. This is the first report of colonic ulcers diagnosed and successfully treated in such circumstances. PMID- 15879051 TI - White retinal vessels. PMID- 15879052 TI - A woman with language disturbance. PMID- 15879053 TI - Hyperglycemia after protein ingestion concurrent with injection of a GLP-1 receptor agonist in rats: a possible role for dietary peptides. AB - Protein ingestion after injection of the glucagon-like peptide-1 receptor agonist Exendin-4 (Ex-4) causes hyperglycemia in rats. The objectives of this study were to determine the components of protein digestion responsible for this effect and to associate it with changes in the concentrations of other metabolites and hormones. Two experiments were conducted. In the first experiment, food-deprived rats were gavaged with intact whey (WP) or albumin protein, their hydrolysates, amino acid mixtures (1 g/2.5 ml), or water 5 min after injection of either PBS or Ex-4 (0.5 microg/rat). Tail vein blood was analyzed for glucose over 2 h. In the second experiment, food-deprived rats were gavaged with WP with or without Ex-4. Groups of conscious rats were killed by decapitation either before, or at selected times after gavage. Plasma concentrations of glucose, amino acids, free fatty acids (FFA), glycerol, insulin, glucagon, and leptin were measured. In experiment 1, blood glucose was higher when intact proteins and protein hydrolysates, but not amino acid mixtures, were given with than without Ex-4 (P < 0.05). In experiment 2, concentrations of glucose, FFA, and the ratio of tyrosine to branched-chain amino acid were higher (P < 0.01), but leptin and essential amino acid concentrations were lower (P < 0.05), and insulin, glucagon, and glycerol were similar when WP was given with or without Ex-4. We conclude that the hyperglycemia caused by the administration of Ex-4 concurrently with dietary protein arises from the action of peptides released during digestion and their interaction with Ex-4 in the regulation of glucose, fatty acid, and amino acid metabolism. PMID- 15879054 TI - Diminished feeding responsiveness to orexin A (hypocretin 1) in aged rats is accompanied by decreased neuronal activation. AB - Orexin A is produced in caudal lateral, posterior, perifornical, and dorsomedial hypothalamic areas. Orexin A in the rostro-dorsal lateral hypothalamic area (rLHa) stimulates feeding and activates several feeding-regulatory brain areas. We hypothesized that aging diminishes feeding and c-Fos-immunoreactivity (c-Fos ir; marker of neuronal activation) response to orexin A. Young (3 mo), middle aged (12 mo), and old (24 mo) male Fischer 344 rLHa-cannulated rats were injected with orexin A (0.5, 1, and 2 nmol). Food intake was measured at 1, 2, and 4 h. c Fos-ir in hypothalamic, limbic, and hindbrain regions was measured in two additional sets of rLHa-orexin A injected rats. In a separate study, orexin A effects on feeding and c-Fos-ir were measured in 6-mo-old rats. Orexin A significantly elevated feeding in rats aged 3, 6, and 12 mo in the 0-1 and 1-2- h time intervals, whereas in old rats this was significant in the 1-2 h time interval only. At 1 h, 6-8 (of 14) brain areas showed elevated c-Fos-ir in response to orexin A in 3- and 6-mo-old rats, but 24-mo-old rats exhibited attenuated or absent c-Fos-ir response in all brain regions except the hypothalamic paraventricular nucleus (PVN) and rostral nucleus of the solitary tract (rNTS). Orexin A did not elevate c-Fos-ir in 3-mo-old rats at 2 h after injection, whereas the PVN and mediodorsal thalamic nucleus (MD) showed elevated c-Fos-ir at 2 h in 24-mo-old rats. These data suggest that delayed and diminished feeding responses in old animals may be due to ineffective neural signaling and implicate the orexin A network as one feeding system affected by aging. PMID- 15879055 TI - ANG III induces expression of inducible transcription factors of AP-1 and Krox families in rat brain. AB - In addition to rapid responses comprising increases in blood pressure, drinking, and stimulation of natriuresis, ANG II induces the expression of transcription factors (TF) in the central nervous system. The ANG II metabolite ANG III (ANG 2 8) has been demonstrated to exert physiological effects similar to those of ANG II. We aimed to determine 1) whether ANG III induces TF expression in the brain, 2) which ANG II (AT) receptor subtype is involved, and 3) whether the two peptides, ANG II and ANG III, differ in their efficacy to stimulate TF expression. ANG II (100 pmol), ANG III (100 pmol), or vehicle was injected into the lateral brain ventricle of conscious rats alone or in combination with the AT(1) receptor antagonist losartan (10 nmol), the AT(2) receptor antagonist PD 123319 (5 nmol), or the aminopeptidase inhibitor amastatin (10 nmol). Similar to ANG II, ANG III induced the expression of c-Fos, c-Jun, and Krox-24 in four brain regions, subfornical organ, median preoptic area, paraventricular nucleus, and supraoptic nucleus of the hypothalamus, with the same efficacy. This effect was AT(1) receptor mediated. Pretreatment with amastatin reduced the expression of TF in response to ANG II, indicating that this expression is partly mediated by ANG III. Interestingly, the AT(2) receptor antagonist PD-123319 alone slightly enhanced the expression of c-Fos, c-Jun, and Krox-24 in different populations of neurons of the paraventricular nucleus. These data indicate that different populations of neurons in the paraventricular nucleus are tonically inhibited by AT(2) receptors under physiological conditions. PMID- 15879056 TI - F-DIO obesity-prone rat is insulin resistant before obesity onset. AB - We previously created a novel F-DIO rat strain derived by crossing rats selectively bred for the diet-induced obesity (DIO) phenotype with obesity resistant Fischer F344 rats. The offspring retained the DIO phenotype through 3 backcrosses with F344 rats but also had exaggerated insulin responses to oral glucose before they became obese on a 31% fat high-energy (HE) diet. Here, we demonstrate that chow-fed rats from the subsequent randomly bred progeny required 57% lower glucose infusions to maintain euglycemia during a hyperinsulinemic clamp in association with 45% less insulin-induced hepatic glucose output inhibition and 80% lower insulin-induced glucose uptake than F344 rats. The DIO phenotype and exaggerated insulin response to oral glucose in the nonobese, chow fed state persisted in the F6 generation. Also, compared with F344 rats, chow-fed F-DIO rats had 68% higher arcuate nucleus proopiomelanocortin mRNA expression which, unlike the increase in F344 rats, was decreased by 26% on HE diet. Further, F-DIO lateral hypothalamic orexin expression was 18% lower than in F344 rats and was increased rather than decreased by HE diet intake. Finally, both maternal obesity and 30% caloric restriction during the third week of gestation produced F-DIO offspring which were heavier and had higher leptin and insulin levels than lean F-DIO dam offspring. Third-gestational week dexamethasone also produced offspring with higher leptin and insulin levels but with lower body weight. Thus F-DIO rats represent a novel and potentially useful model for the study of DIO, insulin resistance, and perinatal factors that influence the development and persistence of obesity. PMID- 15879057 TI - Mechanisms of oleoylethanolamide-induced changes in feeding behavior and motor activity. AB - Oleoylethanolamide (OEA), a lipid synthesized in the intestine, reduces food intake and stimulates lipolysis through peroxisome proliferator-activated receptor-alpha. OEA also activates transient receptor potential vanilloid type 1 (TRPV1) in vitro. Because the anorexigenic effect of OEA is associated with delayed feeding onset and reduced locomotion, we examined whether intraperitoneal administration of OEA results in nonspecific behavioral effects that contribute to the anorexia in rats. Moreover, we determined whether circulating levels of other gut hormones are modulated by OEA and whether CCK is involved in OEA induced anorexia. Our results indicate that OEA reduces food intake without causing a conditioned taste aversion or reducing sodium appetite. It also failed to induce a conditioned place aversion. However, OEA induced changes in posture and reduced spontaneous activity in the open field. This likely underlies the reduced heat expenditure and sodium consumption observed after OEA injection, which disappeared within 1 h. The effects of OEA on motor activity were similar to those of the TRPV1 agonist capsaicin and were also observed with the peroxisome proliferator-activated receptor-alpha agonist Wy-14643. Plasma levels of ghrelin, peptide YY, glucagon-like peptide 1, and apolipoprotein A-IV were not changed by OEA. Finally, antagonism of CCK-1 receptors did not affect OEA-induced anorexia. These results suggest that OEA suppresses feeding without causing visceral illness and that neither ghrelin, peptide YY, glucagon-like peptide 1, apolipoprotein A-IV, nor CCK plays a critical role in this effect. Despite that OEA-induced anorexia is unlikely to be due to impaired motor activity, our data raise a cautionary note in how specific behavioral and metabolic effects of OEA should be interpreted. PMID- 15879058 TI - Foundations of gregariousness in barnacles. 1953. PMID- 15879059 TI - Linking swimming performance, cardiac pumping ability and cardiac anatomy in rainbow trout. AB - We exploited the inherent individual diversity in swimming performance of rainbow trout Oncorhynchus mykiss to investigate the hypothesis that maximum cardiac performance is linked to active metabolic rate (AMR) and critical swimming speed (U crit). Six hundred juveniles (body mass approximately 150 g) were screened using a swimming challenge of 1.2 m s(-1) to identify 'poor swimmers' and 'good swimmers', i.e. the first and last 60 fish to fatigue, respectively. These 120 fish were individually tagged and then reared in common tanks for 9 months, where they grew at similar rates and achieved a similar body mass of approximately 1100 g. Critical swimming speed (U crit) was then measured individually in tunnel respirometers, with simultaneous recordings of cardiac output via a ventral aortic flow probe. The group of individuals that were screened as poor swimmers remained so, with a significantly (27%) lower U crit than good swimmers [89+/-10 cm s(-1) vs 123+/-5 cm s(-1) (mean +/-s.e.m.), respectively, N = 6], a 19% lower AMR (147+/-12 micromol min(-1) kg(-1) vs 181+/-11 micromol min(-1) kg(-1), respectively), and a 30% lower maximum in vivo cardiac output (47.3+/-4.7 ml min( 1) kg(-1) vs 68.0+/-5.2 ml min(-1) kg(-1), respectively). When cardiac performance was compared with an in situ heart preparation, hearts from poor swimmers had a significantly (26%) lower maximum cardiac output (45.9+/-1.9 ml min(-1) kg(-1) vs 56.4+/-2.3 ml min(-1) kg(-1), respectively) and a 32% lower maximum cardiac power output at a high afterload (3.96+/-0.58 mW g(-1) vs 5.79+/ 1.97 mW g(-1), respectively). Cardiac morphology was visualised in vivo by Doppler echography on anaesthetised individual fish and revealed that poor swimmers had a significantly more rounded ventricle (reduced ventricle length to height ratio) compared with good swimmers, which in turn was correlated with fish condition factor. These results provide clear evidence that maximum cardiac performance is linked to AMR and U crit and indicate that a simple screening test can distinguish between rainbow trout with lower active metabolic rate, U crit, maximal cardiac pumping capacity and a more rounded ventricular morphology. These distinguishing traits may have been retained for 9 months despite a common growing environment and growth. PMID- 15879060 TI - Colouration in crab spiders: substrate choice and prey attraction. AB - Australian crab spiders Thomisus spectabilis ambush pollinating insects, such as honeybees (Apis mellifera) on flowers, and can change their body colour between yellow and white. It is traditionally assumed that the spiders change their colour to match the flower colour, thus rendering them cryptic to insect prey. Here, we test this assumption combining state-of-the-art knowledge of bee vision and behavioural experiments. In the field, yellow spiders are only found on yellow daisies (Chrysanthemum frutescens), whereas white spiders are found on yellow and white daisies. These field patterns were confirmed in the laboratory. When given the choice between white and yellow daisies, yellow spiders preferred yellow daisies, whereas white spiders showed only a slight but non-significant preference for white flowers. Thus, T. spectabilis select background colours according to their own body colour. When viewed from a distance, bees use an achromatic signal produced by their green receptors for target detection. Through this visual channel, white spiders on white flowers, and yellow spiders on yellow flowers are virtually undetectable. From a closer distance of a few centimetres, when bees evaluate colour contrast, the combination of spider colour against different flower backgrounds affected the response of honeybees, but not in ways predicted by a classical crypsis/conspicuousness interpretation. Yellow spiders on yellow flowers are not perfectly matched when interpreted through the colour vision of a honeybee. Nevertheless, honeybees showed indifference to the presence of a spider, equally landing on vacant or spider-occupied flowers. Likewise, white spiders are poorly hidden on white flowers, as white spiders reflect ultraviolet light strongly, while white flowers do not. Surprisingly, bees are attracted to this contrast, and significantly more honeybees preferred white flowers occupied by white spiders. White spiders on yellow flowers produce the highest colour contrast and bees again preferred spider-occupied flowers. Yellow spiders on white flowers were the only pairing where bees rejected spider occupied flowers, especially in cases where the contrast between the two was relatively strong. Thus, T. spectabilis select flower colours adaptively in a way that deceives honeybees, or at least does not deter them. PMID- 15879061 TI - Hypoxic responses of Na+/K+ ATPase in trout hepatocytes. AB - Reduction in oxygenation induces inhibition of Na+/K+ ATPase in a number of cells and tissues, including hepatocytes. When not reversed, decrease in Na+/K+ pump activity leads to a gradual Na+ accumulation, cell swelling and death. However, when accompanied by suppression of dissipative cation pathways, it has also been shown to be a beneficial adaptive strategy used by some hypoxia-tolerant species to reduce ATP consumption during prolonged periods of anoxia. This study aims to investigate acute hypoxic responses of the Na+/K+ ATPase in primary cultures of trout hepatocytes. Gradual decrease in oxygenation was followed by an instantaneous transient dose-dependent downregulation of the Na+/K+ ATPase transport activity, but was without an effect on hydrolytic function of the enzyme. Hypoxia-induced inhibition of active K+ influx was reversed spontaneously when hypoxic incubation time exceeded 20 min. The stimulating effect of prolonged hypoxic exposure on the Na+/K+ pump is most probably secondary to hypoxia-induced activation of the Na+/H+ exchanger with the following Na+ accumulation leading to Na+/K+ ATPase activation. Hypoxia-induced inhibition of the Na+/K+ pump was not caused by ATP depletion or global oxidative stress. However, local controlled production of reactive oxygen species seems to play an important role in hypoxia induced regulation of the Na+/K+ ATPase. Treatment of cells with mercaptopropionyl glycine (MPG), a scavenger of OH*-, abolished hypoxia-induced inhibition of the Na+/K+ ATPase. Earlier on we have shown that activation of Na+/H+ exchanger under hypoxic conditions can be opposed by MPG treatment as well. Taken together our results suggest that regulation of both oxygen-sensitive transporters may be accomplished by local changes in free radical production. PMID- 15879062 TI - The ecology of visual pigment tuning in an Australian marsupial: the honey possum Tarsipes rostratus. AB - While most mammals have no more than two types of cone photoreceptor, four species of Australian marsupial have recently been shown to possess three types, and thus have the potential for trichromatic colour vision. Interestingly, the long-wave cones of the honey possum Tarsipes rostratus are tuned to longer wavelengths than those of the other species measured to date. We tested whether the honey possum's long-wave tuning is adaptive for visual tasks associated with its almost unique diet of nectar and pollen. We modelled three tasks: (1) detecting food-rich 'target' flowers against their natural background of foliage or other vegetation; (2) discriminating target flowers from flowers of non-target species; (3) discerning the maturity of the most important target flowers. Initial comparisons of trichromacy vs dichromacy generally favoured the former, but interestingly dichromacy was no disadvantage in some cases. For tuning, we found that overall the honey possum's long-wave tuning is more adaptive than that of the other marsupial species. Nevertheless, the optimal tuning for tasks 1 and 2 would be at longer wavelengths still, implying that a different pressure or constraint operates against a further long-wave shift of the honey possum's L cone tuning. Our data show that a possible ecological pressure may be provided by the third task--the difficult and potentially critical discrimination of the maturity of the animal's major food supply, the flowers of Banksia attenuata. PMID- 15879063 TI - A 3-D kinematic analysis of gliding in a flying snake, Chrysopelea paradisi. AB - Flying snake species (Chrysopelea) locomote through the air despite a lack of appendages or any obvious external morphological specialization for flight. Here photogrammetric techniques were used to investigate C. paradisi's aerial trajectory in three dimensions. Two videocameras arranged in stereo were used to record head, midpoint and vent landmarks on snakes that jumped from a horizontal branch at a height of 9.62 m and landed in an open field. The coordinates of these landmarks were reconstructed in three dimensions and used to analyze patterns of position, glide angle and speed concurrently with changes in body posture in 14 glide sequences from different individuals. C. paradisi's trajectory was composed of a ballistic dive followed by a shallowing phase in which the path became more horizontal; for most glide trials, no equilibrium phase was observed. In the ballistic dive, the snake changed posture from generally straight to a wide 'S' shape in planview and began aerial undulation. Shortly after the ballistic dive, the snake's speed transitioned from an initial acceleration to stable or to a different rate of increase or decrease. Aerial undulation, in which high-amplitude traveling waves pass posteriorly down the body, was a prominent locomotor behavior. In mid-glide, this undulation occurred with the anterior body oriented approximately parallel with the ground and the posterior body cycling up and down in the vertical plane. The body angle of attack for the anterior body for one trial was 20-40 degrees . Snakes traveled a horizontal distance of 10.14+/-2.69 m (mean +/-s.d.) while reaching an airspeed of 10.0+/-0.9 m s(-1), sinking speed of 6.4+/-0.8 m s(-1) and horizontal speed of 8.1+/-0.9 m s(-1). The glide path shallowed at a rate of 20+/-6 degrees s(-1) and reached a minimum glide angle of 28+/-10 degrees , with a minimum recorded glide angle of 13 degrees . C. paradisi are surprisingly good gliders given their unconventional locomotor style, with performance characteristics that rival or surpass more familiar gliding taxa such as flying squirrels. As in other gliders, C. paradisi is potentially capable of using aerial locomotion to move effectively between trees, chase aerial prey, or avoid predators. PMID- 15879064 TI - Effects of size and behavior on aerial performance of two species of flying snakes (Chrysopelea). AB - Aerial locomotion in snakes (genus Chrysopelea) is kinematically distinct from any other type of gliding or powered flight, with prominent, high amplitude body undulations visually dominating the behavior. Because it is not known how flying snakes produce aerodynamic forces in flight, the factors that determine snake flight performance are not clear. In this study, the effects of size and behavior on aerial performance were examined both within a species (C. paradisi) and between two species (C. paradisi and C. ornata), using stepwise multiple regressions to identify relevant variables. Smaller C. paradisi traveled farther than larger snakes at lower sinking speeds, with trajectories that shallowed more quickly and reached lower minimum glide angles. Although wing loading increased faster than expected for isometric size increase, wing loading per se was not responsible for performance differences between large and small snakes. Snakes with higher interactions between relative undulation amplitude and body size transitioned out of the initial acceleration phase at higher airspeeds and sinking speeds, and attained higher maximum airspeeds and horizontal speeds; snakes that used higher average relative amplitudes transitioned out of the initial acceleration phase at higher horizontal speeds. Undulation frequency was not significantly related to any performance variable within C. paradisi and was not significantly different between the two species, suggesting that this variable (in contrast to relative undulation amplitude) may have a minor influence on the aerodynamic mechanism of force production in snake flight. C. paradisi and C. ornata differed significantly in most performance comparisons. C. ornata were more massive than C. paradisi at any given body length and in general exhibited poorer gliding performance than C. paradisi. This study contributes towards understanding how an unconventional body form and kinematics can produce a novel mode of aerial locomotion in a vertebrate glider. PMID- 15879065 TI - Insect egg deposition induces defence responses in Pinus sylvestris: characterisation of the elicitor. AB - Egg deposition by the phytophagous sawfly Diprion pini L. (Hymenoptera, Diprionidae) is known to induce locally and systemically the emission of volatiles in Scots pine (Pinus sylvestris L.) that attract the egg parasitoid Chrysonotomyia ruforum Krausse (Hymenoptera, Eulophidae). The egg parasitoids kill the eggs and thus prevent damage to the plant from feeding sawfly larvae. The elicitor inducing the pine's response is known to be located in the oviduct secretion which the female sawfly applies to the eggs when inserting them into a slit in the pine needle using the sclerotized ovipositor valves. In this study we have characterized the elicitor. The elicitor was still active when isolated from the oviduct and applied directly to slits made in the pine needles. However, as soon as the oviduct secretion was dissolved in Aqua dest. and stored for 3 h at room temperature or kept frozen at -80 degrees C, its activity was lost. In contrast, oviduct secretion kept its eliciting activity, when dissolved in Ringer solution (pH 7.2) both after storage at room temperature and after freezing. The activity of the elicitor vanished after treatment of the oviduct secretion with proteinase K, which destroyed all proteins. This suggests that the elicitor in the oviduct secretion is a peptide or protein, or a component bound to these. SDS PAGE revealed a similar, but not identical protein pattern from hemolymph and oviduct secretion. Hemolymph itself has no eliciting effect. The elicitor in the oviduct secretion is only active when transferred to slit pine needles, since its application on undamaged needles did not induce the emission of attractive volatiles. PMID- 15879066 TI - Appetite-suppressing effects of ammonia exposure in rainbow trout associated with regional and temporal activation of brain monoaminergic and CRF systems. AB - To assess whether the brain's monoaminergic and/or corticotropin-releasing factor (CRF) systems may be involved in mediating the appetite-suppressing effects of high environmental ammonia levels, we exposed rainbow trout to one of four NH4Cl treatments (0, 500, 750, 1000 micromol l(-1)) for 24 or 96 h and monitored changes in food intake, brain serotonin (5-HT) and dopamine (DA) activity, CRF and urotensin I (UI) mRNA levels, and plasma cortisol levels. Food intake decreased in a dose-dependent manner after 24 h of ammonia exposure and partially recovered in all groups after 96 h. Ammonia also elicited dose-dependent increases in serotonergic activity in the hypothalamus (HYP), telencephalon (TEL) and posterior brain (PB). Whereas the increase in serotonergic activity was timed with the 24 h food intake inhibition, TEL and PB serotonergic activity increased after 96 h. In the PB, exogenous ammonia also elicited dose-dependent increases in dopaminergic activity after both 24 and 96 h of exposure. Transient increases in TEL CRF and UI mRNA levels, HYP UI mRNA levels, and plasma cortisol concentrations were evidence that the hypothalamic-pituitary-interrenal (HPI) stress axis was primarily stimulated in the first 24 h of ammonia exposure when food intake was depressed. Overall, the transient nature of the appetite suppression during chronic ammonia exposure, and the time-dependent changes in brain monoaminergic and CRF systems, implicate 5-HT, DA, CRF and UI as potential mediators of the appetite-suppressing effects of ammonia. Among these anorexigenic signals, our results specifically identify hypothalamic 5-HT as a potentially key neurobiological substrate for the regulation of food intake during exposure to high external ammonia concentrations. PMID- 15879067 TI - Timing of praying mantis evasive responses during simulated bat attack sequences. AB - Praying mantids perform evasive maneuvers that vary with the level of danger posed by their bat predators. The vocalization pattern of attacking bats provides cues that mantids can potentially use to decide how and when to respond. Using pulse trains simulating bat attack echolocation sequences, this study determines when in the attack sequence the mantis power dive (its response to high-level threat) occurs and predicts the parameters within the echolocation sequence that are important for eliciting the response. For sequences with a rapid transition from low to high pulse repetition rates (PRRs), the evasive response occurred close to the point during the simulated sequence when the bat would have contacted the mantis. However, the evasive response occurred earlier if the transition was gradual. Regardless of the transition type, the prediction data show that sequences trigger the response when PRRs reach 20-40 pulses s(-1). These results suggest that a bat gradually increasing its PRR could 'tip off' the mantis, enabling it to escape. Attack sequences contain gradual transitions when bats engage in strobing behavior, an echolocation phenomenon that may help the bat perceive the auditory scene. Conversely, bat attack sequences that contain rapid increases in PRR close to the point of capture could circumvent the mantid's auditory defense. Based on these findings, mantids as well as other insects could benefit from having a back-up defense response to offset any advantage the bat gains by rapidly switching from low to high PRRs. PMID- 15879068 TI - Arterial hemodynamics and mechanical properties after circulatory intervention in the chick embryo. AB - Altered blood pressure and flow impact cardiac function during morphogenesis. How the arterial system supports cardiac morphogenesis after circulatory disruptions is not well characterized. We manipulated arterial flow via left atrial ligation (LAL) or arterial load via right vitelline artery ligation (VAL) in Hamburger Hamilton (HH) stage 21 chick embryos. Embryos were reincubated for 1 h (HH21), 14 h (HH24) or 30 h (HH27). At each stage we measured simultaneous dorsal aortic blood pressure and flow, and calculated arterial compliance, impedance and hydraulic power. LAL acutely reduced stroke volume (Vs), cardiac output (Q) and hydraulic power. Arterial pressure was preserved by a compensatory increase in characteristic impedance and decrease in compliance. Impedance parameters and compliance normalized by HH24 and all parameters normalized by HH27. VAL acutely increased arterial resistance. Embryos maintained arterial pressure by decreasing Vs and Q. These parameters remained altered through HH27. In summary, despite the intervention, compensatory alterations in Vs and arterial resistance maintained arterial pressure and fraction of oscillatory power within a narrow range. These results suggest that the maintenance of arterial pressure and circulatory energy efficiency, but not arterial flow, is critical to embryogenesis. PMID- 15879069 TI - The effect of desiccation on water management and compartmentalisation in scorpions: the hepatopancreas as a water reservoir. AB - Scorpions of the Family Buthidae have lower water loss rates (WLR) and enhanced osmoregulatory capacities in comparison with sympatric species of F. Scorpionidae. In this study we followed changes in water content of different body compartments in four scorpion species under prolonged desiccation conditions. The high initial WLR previously reported for Scorpionidae result in rapid depletion of body water stores. A significant decrease in total body water content of Scorpionidae was recorded following loss of only 5% of initial mass, whereas no such decrease was recorded for Buthidae following severe desiccation. When desiccated, scorpions lose water primarily from the hepatopancreas, while haemolymph volume is more tightly regulated. However, the haemolymph volume of Scorpionidae decreases as a result of depletion of hepatopancreas water stores following severe desiccation. The increasing lipid fraction in the hepatopancreas of Scorpionidae during desiccation suggests that depletion of body water stores may induce enhanced catabolism of carbohydrates, which may contribute to volume regulation by making initially glycogen-bound water available to the desiccating scorpion. PMID- 15879070 TI - Changes in gut and Malpighian tubule transport during seasonal acclimatization and freezing in the gall fly Eurosta solidaginis. AB - Since few studies have examined cold tolerance at the organ level in insects, our primary objective was to characterize the functional responses of the gut and Malpighian tubules (MT) to seasonal acclimatization, chilling and freezing in larvae of the goldenrod gall fly Eurosta solidaginis Fitch (Diptera, Tephritidae). From September to December, hemolymph osmolality (455-926 mOsmol kg l(-1)) and freezing tolerance increased markedly in field-collected larvae. Chlorophenol Red was readily transported into the lumen of the foregut, the posterior portion of the midgut, the ureter, the proximal region of the anterior pair of MT, and entire posterior pair of MT. Ouabain and KCN inhibited transport of Chlorophenol Red in the gut and MT. Transport was readily detected at 0 degrees C and the rate of transport was directly related to temperature. The rate of fluid transport by the MT decreased steadily from a monthly high in September (10.7+/-0.8 nl min(-1) for the anterior pair; 12.7+/-1.0 nl min(-1) for the posterior pair) until secretion was no longer detectable in December; this decrease parallels entry into diapause for this species. Even in larvae that died following freezing for 40 days at -20 degrees C, individual organ function was retained to a limited extent. Through the autumn, cholesterol concentrations in the hemolymph increased nearly fourfold. In contrast, the ratio of cholesterol to protein content (nmol mg l(-1)) in the MT membrane remained relatively constant (22 approximately 24 nmol mg l(-1) protein) during this period. Freezing of larvae for 20 days at -20 degrees C caused a significant decrease in cholesterol levels in the hemolymph and the MT membranes compared to unfrozen controls. These results suggest that cholesterol plays a role in seasonal cold hardening and freeze tolerance in insects. PMID- 15879071 TI - Contractile properties of mouse single muscle fibers, a comparison with amphibian muscle fibers. AB - Single fibers, 25-40 microm wide and 0.5-0.7 mm long, were isolated from the flexor digitorum brevis muscle of the mouse. Force and movement were recorded (21 27 degrees C) from the fiber as a whole and, in certain experiments, from a short marked segment that was held at constant length by feedback control. The maximum tetanic force, 368+/-57 kN/m2 (N = 10), was not significantly different from that recorded in frog muscle fibers at equal temperature. However, the rising phase of the tetanus was considerably slower in the mammalian fibers, 202+/-20 ms (N = 17) being required to reach 90% of maximum tetanic force as compared with 59+/-4 ms (N = 20) in the frog muscle fibers. Similar to the situation in frog muscle fibers, the force-velocity relation exhibited two distinct curvatures located on either side of a breakpoint near 80% of the isometric force. Maximum speed of shortening was 4.0+/-0.3 fiber lengths s(-1) (N = 6). The relationship between tetanic force and sarcomere length was studied between 1.5 and 4.0 microm sarcomere spacings, based on length-clamp recordings that were free of 'tension creep'. There was a flat maximum (plateau) of the length-tension relation between approximately 2.0 and 2.4 microm sarcomere lengths. The descending limb of the length-tension relation (linear regression) intersected the length axis (zero force) at 3.88 microm and reached maximum force at 2.40 microm sarcomere length. The slope of the descending limb is compatible with a thick filament length of 1.63 microm and an average thin filament length of 1.10 microm. These values accord well with recent electron microscope measurements of myofilament length in mammalian muscle. PMID- 15879072 TI - Genomic sequences encoding two types of medaka hemopexin-like protein Wap65, and their gene expression profiles in embryos. AB - Medaka genomic BAC clones, which contained two types of medaka hemopexin-like protein gene (Wap65), mWap65-1 and mWap65-2, were screened and their genomic sequences were determined by the shotgun strategy. The exon-intron organizations were highly conserved between both mWap65s and human hemopexin genes. The 5' flanking regions of mWap65-1 and mWap65-2 contained various putative transcription factor binding sites including elements for developmental regulation. The expression patterns of mWap65s during embryonic development were examined by quantitative real-time PCR, demonstrating that both mWap65 transcripts were observed in early embryonic stages, but their expression patterns were different. Interestingly, in situ hybridization revealed that mWap65-2 transcripts were restricted to liver, whereas mWap65-1 transcripts were detected along the edge of pectoral fin buds and the median fin fold of tail buds in embryos at stage 32. Furthermore, we generated transgenic medaka expressing GFP driven by mWap65-1 and mWap65-2 promoters and observed GFP expression patterns during ontogeny. Although localizations of GFP varied among individuals, embryos uniformly expressed GFP 1 day after injection of mWap65-1-hrGFP and mWap65-2-hrGFP constructs, suggesting that mWap65-1 and mWap65-2 promoters were activated in very early stages. The differences between mWap65-1 and mWap65-2 in their expression profiles indicate their distinct roles during ontogeny. PMID- 15879073 TI - Cardiac preload and venous return in swimming sea bass (Dicentrarchus labrax L.). AB - Cardiac preload (central venous pressure, P(CV), mean circulatory filling pressure (MCFP), dorsal aortic blood pressure (P(DA)) and relative cardiac output (Q) were measured in sea bass (Dicentrarchus labrax) at rest and while swimming at 1 and 2 BL s(-1). MCFP, an index of venous capacitance and the upstream venous pressure driving the return of venous blood to the heart, was measured as the plateau in Pcv during ventral aortic occlusion. Compared with resting values, swimming at 1 and 2 BL s(-1) increased Q (by 15+/-1.5 and 38+/-6.5%, respectively), P(CV) (from 0.11+/-0.01 kPa to 0.12+/-0.01 and 0.16+/-0.02 kPa, respectively), MCFP (from 0.27+/-0.02 kPa to 0.31+/-0.02 and 0.40+/-0.04 kPa, respectively) and the calculated pressure gradient for venous return (DeltaP(V), from 0.16+/-0.01 kPa to 0.18+/-0.02 and 0.24+/-0.02 kPa, respectively), but not P(DA). In spite of an increased preload, the increase in Q was exclusively mediated by an increased heart rate (f(H), from 80+/-4 beats min(-1) to 88+/-4 and 103+/-3 beats min(-1), respectively), and stroke volume (Vs) remained unchanged. Prazosin treatment (1 mg kg(-1) Mb) abolished pressure and flow changes during swimming at 1 BL s(-1), but not 2 BL s(-1), indicating that other control systems besides an alpha-adrenoceptor control are involved. This study is the first to address the control of venous capacitance in swimming fish. It questions the generality that increased Q during swimming is regulated primarily through Vs and shows that an increased cardiac filling pressure does not necessarily lead to an increased Vs in fish, but may instead compensate for a reduced cardiac filling time. PMID- 15879074 TI - Quasistatic and continuous dynamic characterization of the mechanical properties of silk from the cobweb of the black widow spider Latrodectus hesperus. AB - Spider silks are among the strongest and toughest known materials, but investigation of these remarkable properties has been confined largely to orb weaving spiders. We investigated the mechanical performance of silk from the cobweb-weaving spider Latrodectus hesperus. Both silk from the scaffolding region of the web and sticky gumfooted capture lines had material properties similar to the major ampullate silk that orb weavers use as the framework for their orb webs. Major ampullate fibers obtained from anaesthetized Latrodectus spiders were similar, but exhibited increased stiffness and reduced extensibility. Novel continuous dynamic analysis of the silks revealed that the loss tangent (tandelta) increased rapidly during the first 2-3% of extension and reached a maximum near the yield point of fibers. The loss tangent then rapidly declined at an ever-decreasing rate until failure. We suggest that these data support molecular models for the mechanics of spider silk. We also demonstrate that the addition of sticky aggregate glue to the ends of the gumfooted lines modulates their mechanical performance--reducing stiffness and increasing extensibility. The storage modulus of viscid regions of the gumfooted lines was much lower than dry regions. This may be explained by disruption of hydrogen bonding within the amorphous regions of the fibers due to hydration from the glue. PMID- 15879075 TI - Cytoplasmic carbonic anhydrase isozymes in rainbow trout Oncorhynchus mykiss: comparative physiology and molecular evolution. AB - It is well established that the gills of teleost fish contain substantial levels of cytoplasmic carbonic anhydrase (CA), but it is unclear which CA isozyme(s) might be responsible for this activity. The objective of the current study was to determine if branchial CA activity in rainbow trout was the result of a general cytoplasmic CA isozyme, with kinetic properties, tissue distribution and physiological functions distinct from those of the red blood cell (rbc)-specific CA isozyme. Isolation and sequencing of a second trout cytoplasmic CA yielded a 780 bp coding region that was 76% identical with the trout rbc CA (TCAb), although the active sites differed by only 1 amino acid. Interestingly, phylogenetic analyses did not group these two isozymes closely together, suggesting that more fish species may have multiple cytoplasmic CA isozymes. In contrast to TCAb, the second cytoplasmic CA isozyme had a wide tissue distribution with high expression in the gills and brain, and lower expression in many tissues, including the red blood cells. Thus, unlike TCAb, the second isozyme lacks tissue specificity and may be expressed in the cytoplasm of all cells. For this reason, it is referred to hereafter as TCAc (trout cytoplasmic CA). The inhibitor properties of both cytoplasmic isozymes were similar (Ki acetazolamide 1.21+/-0.18 nmol l(-1) and 1.34+/-0.10 nmol l(-1) for TCAc and TCAb, respectively). However, the turnover of TCAb was over three times greater than that of TCAc (30.3+/-5.83 vs 8.90+/-1.95 e4 s(-1), respectively), indicating that the rbc-specific CA isoform was significantly faster than the general cytoplasmic isoform. Induction of anaemia revealed differential expression of the two isozymes in the red blood cell; whereas TCAc mRNA expression was unaffected, TCAb mRNA expression was significantly increased by 30- to 60-fold in anaemic trout. PMID- 15879076 TI - Sources of mechanical power for uphill running in humans. AB - During uphill running limb muscles must perform net mechanical work to increase the body's potential energy, while during level running the net mechanical work required is negligible as long as speed is constant. The increased demands for work as running incline increases might be met by an increase in power output at all joints, or only a subset of joints. We used inverse dynamics to determine which joints modulate net work output in humans running uphill. We measured joint kinematics and ground reaction force during moderate speed running at 0 degrees , 6 degrees and 12 degrees inclines. Muscle force, joint power and work per step were determined at the ankle, knee and hip using inverse dynamics calculations. We found that virtually all of the increase in work output with increasing incline resulted from increases in net work done at the hip (-0.25+/-0.23 J kg( 1), level, vs 0.88+/-0.10 J kg(-1), 12 degrees incline), while the knee and ankle performed similar functions at all inclines. The increase in work output at the hip resulted primarily from a large increase in average net muscle moment during stance (2.07+/-17.84 Nm, level, vs 87.30+/-13.89 Nm, 12 degrees incline); joint excursion increased by only 20% (41.22+/-3.41 degrees , level, vs 49.22+/-2.35 degrees , 12 degrees incline). The increase in hip muscle moment and power was associated with a poorer mechanical advantage for producing force against the ground. The increase in hip moment with running incline allows for the production of the power necessary to lift the body. This power may be developed by hip extensors or by transfer of power from muscles at other joints via biarticular muscles. PMID- 15879077 TI - Physiological control of diving behaviour in the Weddell seal Leptonychotes weddelli: a model based on cardiorespiratory control theory. AB - Despite being obligate air breathers, many species of marine mammal are capable of spending most of their lives submerged in water. How they do this has been a subject of intense interest to physiologists for over a century, yet we still do not have a detailed understanding of the physiological mechanisms underlying this behaviour. What are the proximate mechanisms that trigger the 'decisions' to submerge and return to the surface? The present study proposes a model intended to address this question, based on fundamental concepts of cardiorespiratory control. Two basic hypotheses are examined by computer simulation, using a mathematical model of the mammalian cardiorespiratory control system with parameter values for an adult Weddell seal: (1) that the control of diving can be considered to be a respiratory control problem, and (2) that dives are initiated and maintained by disfacilitation of respiratory drive, not inhibition. Computer simulations confirmed the plausibility of these hypotheses. Simulated diving behaviour and physiological responses (ventilation, cardiac output, blood and tissue gas tensions) were consistent with published data from freely diving Weddell seals. Dives up to the estimated aerobic dive limit (ADL, 18-25 min) could be simulated without the need for active inhibition of breathing in this model. This theoretical analysis suggests that the most important physiological adjustments occur during the surface interval phase of the dive cycle and include hyperventilation accompanied by high cardiac output, appropriate regulation of cerebral blood flow and central chemoreceptor threshold shifts. During dives, cardiac output, distribution of peripheral blood flow, splenic contraction and peripheral chemoreflex drives were found to modulate physiological and behavioural responses, but were not essential for simulated dives to occur. The main conclusion from this study is that the central chemoreceptor may be an important mechanism involved in the regulation of diving behaviour, implying that CO2, not O2, is the key regulatory variable in this model. This model includes and extends the ADL concept and suggests an explicit mechanism by which the respiratory control system may play a central role in the regulation of diving behaviour. It is likely that respiratory mechanisms are an important component of a hierarchical behavioural control system and further studies are required to test the qualitative and quantitative validity of the model. PMID- 15879078 TI - Chronic and acute ammonia toxicity in mudskippers, Periophthalmodon schlosseri and Boleophthalmus boddaerti: brain ammonia and glutamine contents, and effects of methionine sulfoximine and MK801. AB - The objective of this study was to elucidate if chronic and acute ammonia intoxication in mudskippers, Periophthalmodon schlosseri and Boleophthalmus boddaerti, were associated with high levels of ammonia and/or glutamine in their brains, and if acute ammonia intoxication could be prevented by the administration of methionine sulfoximine [MSO; an inhibitor of glutamine synthetase (GS)] or MK801 [an antagonist of N-methyl D-aspartate type glutamate (NMDA) receptors]. For P. schlosseri and B. boddaerti exposed to sublethal concentrations (100 and 8 mmol l(-1) NH4Cl, respectively, at pH 7.0) of environmental ammonia for 4 days, brain ammonia contents increased drastically during the first 24 h, and they reached 18 and 14.5 micromol g(-1), respectively, at hour 96. Simultaneously, there were increases in brain glutamine contents, but brain glutamate contents were unchanged. Because glutamine accumulated to exceptionally high levels in brains of P. schlosseri (29.8 micromol g(-1)) and B. boddaerti (12.1 micromol g(-1)) without causing death, it can be concluded that these two mudskippers could ameliorate those problems associated with glutamine synthesis and accumulation as observed in patients suffering from hyperammonemia. P. schlosseri and B. boddaerti could tolerate high doses of ammonium acetate (CH3COONH4) injected into their peritoneal cavities, with 24 h LC50 of 15.6 and 12.3 micromol g(-1) fish, respectively. After the injection with a sublethal dose of CH3COONH4 (8 micromol g(-1) fish), there were significant increases in ammonia (5.11 and 8.36 micromol g(-1), respectively) and glutamine (4.22 and 3.54 micromol g(-1), respectively) levels in their brains at hour 0.5, but these levels returned to normal at hour 24. By contrast, for P. schlosseri and B. boddaerti that succumbed within 15-50 min to a dose of CH3COONH4 (15 and 12 micromol g(-1) fish, respectively) close to the LC50 values, the ammonia contents in the brains reached much higher levels (12.8 and 14.9 micromol g(-1), respectively), while the glutamine level remained relatively low (3.93 and 2.67 micromol g(-1), respectively). Thus, glutamine synthesis and accumulation in the brain was not the major cause of death in these two mudskippers confronted with acute ammonia toxicity. Indeed, MSO, at a dosage (100 microg g(-1) fish) protective for rats, did not protect B. boddaerti against acute ammonia toxicity, although it was an inhibitor of GS activities from the brains of both mudskippers. In the case of P. schlosseri, MSO only prolonged the time to death but did not reduce the mortality rate (100%). In addition, MK801 (2 microg g(-1) fish) had no protective effect on P. schlosseri and B. boddaerti injected with a lethal dose of CH3COONH4, indicating that activation of NMDA receptors was not the major cause of death during acute ammonia intoxication. Thus, it can be concluded that there are major differences in mechanisms of chronic and acute ammonia toxicity between brains of these two mudskippers and mammalian brains. PMID- 15879079 TI - Nitric oxide modulates peristaltic muscle activity associated with fluid circulation in the sea pansy Renilla koellikeri. AB - Nitric oxide (NO) is a well-known regulator of vascular activities in vertebrates and it has also been implicated as a vasodilatatory agent in a cephalopod. In the sea pansy Renilla koellikeri, an octocorallian representative of the most basal animals with a nervous system, we investigated the role of NO in peristalsis, an activity that moves body fluids through the coelenteron (gastrovascular cavity) of the polyps across the colony. NO donors increased the amplitude of peristaltic contractions and increased tonic contractions in relaxed preparations, but caused a relaxation of basal tension in contracted preparations. The NO synthase (NOS) inhibitors L-NAME (N(omega)-nitro-L-arginine methyl ester) and 7-nitroindazole reduced the amplitude of peristaltic contractions and lowered basal tension. In contrast, aminoguanidine, a specific inhibitor of inducible NOS, increased the amplitude but reduced the rate of peristalsis. Zaprinast, a cGMP-specific phosphodiesterase inhibitor, decreased the amplitude of peristaltic contractions, a decrease that was amplified by dibutyryl cGMP. In contrast, the inhibitor of soluble guanylyl cyclase ODQ (1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one) enhanced peristalsis. Putative NOS-containing neurons, revealed by NADPH diaphorase activity and citrulline immunohistochemistry, were observed in the basiectoderm at the base of the autozooid polyp tentacles and in a nerve-net around the oral disc. Their neurites ran up the tentacles and down to the polyp body wall, crossing from the ectoderm through the mesoglea and into the endoderm musculature where musculo-epithelial cells were also reactive. These data suggest that two distinct nitrergic pathways, one of which is mediated by cGMP, regulate peristalsis and muscle tone in the sea pansy and that these pathways may involve NOS-containing ectodermal neurons and musculo-epithelial cells. PMID- 15879080 TI - License to heal: bidirectional interaction of antigen-specific regulatory T cells and tolerogenic APC. AB - Naturally occurring CD4(+)CD25(+) regulatory T (T(R)) cells, a component of the innate immune response, which play a key role in the maintenance of self tolerance, have become the focus of numerous studies over the last decade. These cells inhibit the immune response in an Ag-nonspecific manner, interacting with other T cells. Much less is known about adaptive T(R) cells, which develop in response to chronic antigenic stimulation, and act directly on professional and nonprofessional APC, rendering them tolerogenic and able to elicit the differentiation of CD8(+) and CD4(+) T cells with suppressive activity. In this review, we will discuss data pertaining to the bidirectional interaction between Ag-specific T(R) with APC and their clinical relevance. PMID- 15879081 TI - Cutting edge: CD95 maintains effector T cell homeostasis in chronic immune activation. AB - The elimination of activated T cells is important to maintain homeostasis and avoid immunopathology. CD95 (Fas/APO-1) has been identified as a death mediator for activated T cells in vitro but the function of CD95 in death of mature T cells in vivo is still controversial. Here we show that triggering of the costimulatory TNF receptor family member CD27 sensitized T cells for CD95-induced apoptosis. CD95-deficient (lpr/lpr) T cells massively expanded and differentiated into IFN-gamma-secreting effector cells in transgenic mice that constitutively express the CD27 ligand, CD70. Concomitantly, CD95-deficient CD70 transgenic mice became moribund by 4 wk of age with severe liver pathology and bone marrow failure. These findings establish that CD95 is a critical regulator of effector T cell homeostasis in chronic immune activation. PMID- 15879082 TI - Cutting edge: the acquisition of TLR tolerance during malaria infection impacts T cell activation. AB - An effective immune response to infection requires control of pathogen growth while minimizing inflammation-associated pathology. During malaria infection, this balance is particularly important. Murine malaria is characterized by early production of proinflammatory cytokines, which declines in the face of continuing parasitemia. The mechanism by which this occurs remains poorly understood. In this study, we investigated the role of dendritic cells (DCs) in regulating pro- and anti-inflammatory cytokine responses. As malaria infection progresses, DCs become refractory to TLR-mediated IL-12 and TNF-alpha production, while increasing their ability to produce IL-10 and retaining the capacity for activation of naive T cells. IL-12-secreting DCs from early infection stimulate an IFN-gamma-dominated T cell response, whereas IL-10-secreting DCs from later stages induce an IL-10-dominated T cell response. We suggest that phenotypic changes in DCs during Plasmodium yoelii infection represent a mechanism of controlling host inflammation while maintaining effective adaptive immunity. PMID- 15879083 TI - Cutting edge: emergence of CD127high functionally competent memory T cells is compromised by high viral loads and inadequate T cell help. AB - In this report we have inspected whether difficulties in controlling viral infections negatively impacts the generation of CD127(high) memory T cells. Using both MHC class I and II tetramers we reveal that CD127(low) T cells are not necessarily rapidly deleted but can persist in a pseudoeffector state in which they display the hallmarks of activated effector cells but are functionally inferior. CD127(high) cells can, however, emerge if the infection is contained. We also show that in the absence of CD4 T cell help significant populations of CD127(high) CD8 T cells fail to emerge. Analyses of cytokine-producing activities by both mouse and human CD8 T cells further document that the extended maintenance of T cells in a CD127(low) state has functional consequences which manifest as an impairment of IL-2 production. PMID- 15879084 TI - Cutting edge: the SLAM family receptor Ly108 controls T cell and neutrophil functions. AB - Ly108, a glycoprotein of the signaling lymphocytic activation molecule family of cell surface receptors expressed by T, B, NK, and APCs has been shown to have a role in NK cell cytotoxicity and T cell cytokine responses. In this study, we describe that CD4(+) T cells from mice with a targeted disruption of exons 2 and 3 of Ly108 (Ly108(DeltaE2+3)) produce significantly less IL-4 than wild-type CD4(+) cells, as judged by in vitro assays and by in vivo responses to cutaneous infection with Leishmania mexicana. Surprisingly, neutrophil functions are controlled by Ly108. Ly108(DeltaE2+3) mice are highly susceptible to infection with Salmonella typhimurium, bactericidal activity of Ly108(DeltaE2+3) neutrophils is defective, and their production of IL-6, IL-12, and TNF-alpha is increased. The aberrant bactericidal activity by Ly108(DeltaE2+3) neutrophils is a consequence of severely reduced production of reactive oxygen species following phagocytosis of bacteria. Thus, Ly108 serves as a regulator of both innate and adaptive immune responses. PMID- 15879085 TI - Cutting edge: re-evaluating the in vivo cytokine responses of CD8+ T cells during primary and secondary viral infections. AB - Virus-specific CD8(+) T cells produce IFN-gamma after Ag contact and, in the absence of this cytokine, the host often cannot eradicate infection. However, our ability to identify cells that are actively expressing this critical effector function in vivo is limited, because the protein is rapidly secreted. In this study, we describe a simple approach that circumvents the need for ex vivo Ag stimulation and allows the enumeration of CD8(+) T cells that are actively synthesizing IFN-gamma in vivo during primary and secondary virus infections. The proportion of Ag-specific primary CD8(+) T cells producing IFN-gamma peaks at 5 days postinfection, when the T cell population is still expanding exponentially. In vivo IFN-gamma synthesis by memory cells is explosive, peaking at approximately 12 h after secondary infection and terminating hours thereafter. This technique will be useful when evaluating in vivo immune cell activity in many situations, including a variety of noninfectious (e.g., autoimmune) diseases. PMID- 15879086 TI - Effects of cytokines on acetylcholine receptor expression: implications for myasthenia gravis. AB - Myasthenia gravis is an autoimmune disease associated with thymic pathologies, including hyperplasia. In this study, we investigated the processes that may lead to thymic overexpression of the triggering Ag, the acetylcholine receptor (AChR). Using microarray technology, we found that IFN-regulated genes are more highly expressed in these pathological thymic tissues compared with age- and sex-matched normal thymus controls. Therefore, we investigated whether proinflammatory cytokines could locally modify AChR expression in myoid and thymic epithelial cells. We found that AChR transcripts are up-regulated by IFN-gamma, and even more so by IFN-gamma and TNF-alpha, as assessed by real-time RT-PCR, with the alpha-AChR subunit being the most sensitive to this regulation. The expression of AChR protein was increased at the cytoplasmic level in thymic epithelial cells and at the membrane in myoid cells. To examine whether IFN-gamma could influence AChR expression in vivo, we analyzed AChR transcripts in IFN-gamma gene knock-out mice, and found a significant decrease in AChR transcript levels in the thymus but not in the muscle, compared with wild-type mice. However, up-regulation of AChR protein expression was found in the muscles of animals with myasthenic symptoms treated with TNF-alpha. Altogether, these results indicate that proinflammatory cytokines influence the expression of AChR in vitro and in vivo. Because proinflammatory cytokine activity is evidenced in the thymus of myasthenia gravis patients, it could influence AChR expression and thereby contribute to the initiation of the autoimmune anti-AChR response. PMID- 15879087 TI - TGF-beta 1 uses distinct mechanisms to inhibit IFN-gamma expression in CD4+ T cells at priming and at recall: differential involvement of Stat4 and T-bet. AB - TGF-beta1 plays a critical role in restraining pathogenic Th1 autoimmune responses in vivo, but the mechanisms that mediate TGF-beta1's suppressive effects on CD4(+) T cell expression of IFN-gamma expression remain incompletely understood. To evaluate mechanisms by which TGF-beta1 inhibits IFN-gamma expression in CD4(+) T cells, we primed naive wild-type murine BALB/c CD4(+) T cells in vitro under Th1 development conditions in the presence or the absence of added TGF-beta1. We found that the presence of TGF-beta1 during priming of CD4(+) T cells suppressed both IFN-gamma expression during priming as well as the development of Th1 effector cells expressing IFN-gamma at a recall stimulation. TGF-beta1 inhibited the development of IFN-gamma-expressing cells in a dose dependent fashion and in the absence of APC, indicating that TGF-beta1 can inhibit Th1 development by acting directly on the CD4(+) T cell. During priming, TGF-beta1 strongly inhibited the expression of both T-bet (T box expressed in T cells) and Stat4. We evaluated the importance of these two molecules in the suppression of IFN-gamma expression at the two phases of Th1 responses. Enforced expression of T-bet by retrovirus prevented TGF-beta1's inhibition of Th1 development, but did not prevent TGF-beta1's inhibition of IFN-gamma expression at priming. Conversely, enforced expression of Stat4 partly prevented TGF-beta1's inhibition of IFN-gamma expression during priming, but did not prevent TGF beta1's inhibition of Th1 development. These data show that TGF-beta1 uses distinct mechanisms to inhibit IFN-gamma expression in CD4(+) T cells at priming and at recall. PMID- 15879088 TI - CD4+CD25+ regulatory T cells attenuate the phosphatidylinositol 3-kinase/Akt pathway in antigen-primed immature CD8+ CTLs during functional maturation. AB - This study was designed to determine the role of CD25(+)CD4(+) regulatory T (Tr) cells in CTL maturation and effector functions using a murine CTL line and in vitro MLC. Tr cells inhibited CTL functional maturation, but had no effect on CTL effector functions. In CD4(+) responder T cell-depleted MLC supplemented with IL 2, Tr cells suppressed mature CTL generation only when added within the first 2 days of culture. Tr cells down-regulated levels of active Akt, but not STAT5 or ZAP70 in Ag-primed immature CTLs. Down-regulation of active Akt was accompanied by a reduction in CTL cell size and IL-2Ralpha expression. In Tr cell-depleted MLC, CTLs were generated that exhibited high levels of nonspecific cytotoxicity. Our in vitro findings suggest that Tr cells regulate functional CTL maturation to generate optimal Ag-specific immune responses through the control of the PI3K/Akt pathway. PMID- 15879089 TI - Intravenous infusion of syngeneic apoptotic cells by photopheresis induces antigen-specific regulatory T cells. AB - The basis of extracorporeal photopheresis is the reinfusion of leukocytes previously exposed to 8-methoxypsoralen (8-MOP) and UVA radiation. It has been approved for the palliative treatment of cutaneous T cell lymphoma and has reported benefits in autoimmune diseases, transplant rejection, and graft-vs-host disease. However, the underlying mechanism of photopheresis remains unresolved. Because UVB radiation can cause immune tolerance via induction of regulatory T cells, we studied whether photopheresis exerts a similar effect extracorporeally. Therefore, we established a model of photopheresis using a murine model of contact hypersensitivity. Splenocytes and lymph node cells of mice that were sensitized with dinitrofluorobenzene were exposed to 8-MOP plus UVA in vitro. Intravenous injection of these cells into naive mice caused inhibition of a hapten immune response, which was lost upon depletion of CD11c(+) cells but not T cells. Mice that received untreated cells or cells exposed to UVA or 8-MOP alone were not affected. Inhibition was cell-mediated and Ag-specific as demonstrated by transfer of tolerance from the primary recipients into naive animals, which could, however, properly respond to the unrelated hapten oxazolone. Transfer activity was lost when cells were depleted of CD4(+) or CD25(+) subpopulations. These data suggest that photopheresis exerts its immunomodulatory effects via the induction of Ag-specific regulatory T cells. PMID- 15879090 TI - Identification of Grb2 as a novel binding partner of the signaling lymphocytic activation molecule-associated protein binding receptor CD229. AB - Ag recognition by the TCR determines the subsequent fate of the T cell and is regulated by the involvement of other cell surface molecules, termed coreceptors. CD229 is a lymphocyte cell surface molecule that belongs to the CD150 family of receptors. Upon tyrosine phosphorylation, CD229 recruits various signaling molecules to the membrane. One of these molecules is the signaling lymphocytic activation molecule-associated protein, of which a deficiency leads to the X linked lymphoproliferative syndrome. We report that CD229 interacts in a phosphorylation-dependent manner with Grb2. We mapped this interaction showing that the Src homology 2 domain of Grb2 and the tyrosine residue Y606 in CD229 are required for CD229-Grb2 complex formation. The Grb2 motif in the cytoplasmic tail of CD229 is distinct and independent from the two tyrosines required for efficient signaling lymphocytic activation molecule-associated protein recruitment. CD229, but not other members of the CD150 family, directly bound Grb2. We also demonstrate that CD229 precipitates with Grb2 in T lymphocytes after pervanadate treatment, as well as CD229 or TCR ligation. Interestingly, the CD229 mutant lacking the Grb2 binding site is not internalized after CD229 engagement with specific Abs. Moreover, a dominant negative form of Grb2 (containing only Src homology 2 domain) impaired CD229 endocytosis. Unexpectedly, Erk phosphorylation was partially inhibited after activation of CD229 plus CD3. Consistent with this, CD229 ligation partially inhibited TCR signaling in peripheral blood cells and CD229-Jurkat cells transfected with the 3XNFAT luciferase reporter construct. Altogether, the data suggest a model whereby CD229 ligation attenuates TCR signaling and Grb2 recruitment to CD229 controls its rate of internalization. PMID- 15879091 TI - TGF-beta 1 inhibits mast cell Fc epsilon RI expression. AB - Mast cell activation through the high affinity IgE receptor (FcepsilonRI) is a critical component of atopic inflammation. The cytokine TGF-beta1 has been shown to inhibit IgE-dependent mast cell activation, possibly serving to dampen mast cell-mediated inflammatory responses. We present proof that TGF-beta1 inhibits mast cell FcepsilonRI expression through a reversible pathway that diminishes protein, but not mRNA, expression of the FcepsilonRI subunit proteins alpha, beta, and gamma. The stability of the expressed proteins and the assembled cell surface complex was unaltered by TGF-beta1 treatment. However, TGF-beta1 decreased the rate of FcepsilonRI beta-chain synthesis, arguing that this inhibitory cytokine exerts its effects at the level of mRNA translation. TGF beta1 consistently diminished FcepsilonRI expression on cultured human or mouse mast cells as well as freshly isolated peritoneal mast cells. The related cytokines, TGF-beta2 and TGF-beta3, had similar effects. We propose that TGF beta1 acts as a negative regulator of mast cell function, in part by decreasing FcepsilonRI expression. PMID- 15879092 TI - Multiple costimulatory modalities enhance CTL avidity. AB - Recent studies in both animal models and clinical trials have demonstrated that the avidity of T cells is a major determinant of antitumor and antiviral immunity. In this study, we evaluated several different vaccine strategies for their ability to enhance both the quantity and avidity of CTL responses. CD8(+) T cell quantity was measured by tetramer binding precursor frequency, and avidity was measured by both tetramer dissociation and quantitative cytolytic function. We have evaluated a peptide, a viral vector expressing the Ag transgene alone, with one costimulatory molecule (B7-1), and with three costimulatory molecules (B7-1, ICAM-1, and LFA-3), with anti-CTLA-4 mAb, with GM-CSF, and combinations of the above. We have evaluated these strategies in both a foreign Ag model using beta-galactosidase as immunogen, and in a "self" Ag model, using carcinoembryonic Ag as immunogen in carcinoembryonic Ag transgenic mice. The combined use of several of these strategies was shown to enhance not only the quantity, but, to a greater magnitude, the avidity of T cells generated; a combination strategy is also shown to enhance antitumor effects. The results reported in this study thus demonstrate multiple strategies that can be used in both antitumor and antiviral vaccine settings to generate higher avidity host T cell responses. PMID- 15879093 TI - IFN regulatory factor-2 deficiency revealed a novel checkpoint critical for the generation of peripheral NK cells. AB - NK cell development is far less understood compared with that of T and B cells despite the critical importance of NK cells in innate immunity. Mice lacking the transcription factor IFN regulatory factor-2 (IRF-2) are known to exhibit NK cell deficiency. However, the role of IRF-2 in NK cell development has remained unclear. In this study we found that NK cell deficiency in the periphery in IRF-2 deficient mice was due to selective loss of mature NK cells, but not to maturation arrest, and NK cells in these mice exhibited very immature surface phenotypes (CD11b(low)Dx5(low)) with highly compromised NK receptor expression. In contrast, IRF-2-deficient NK cells in bone marrow (BM) showed relatively mature phenotypes (CD11b(low)Dx5(high)) with less compromised NK receptor repertoire. Furthermore, BM NK cells in IRF-2-deficient mice were found to proliferate almost normally, but underwent accelerated apoptosis. These observations indicated that NK cell maturation could advance up to a late, but not the final, stage in the BM, whereas these cells were incapable of contributing to the peripheral NK cell pool due to premature death in the absence of IRF-2. In contrast, NK cell numbers and Ly49 expression were much more severely reduced in BM in IL-15-deficient mice than in IRF-2(-/-) mice. The differential peripheral and central NK cell deficiencies in IRF-2(-/-) mice thus revealed a novel late checkpoint for NK cell maturation, distinct from the early IL-15-dependent expansion stage. PMID- 15879094 TI - CD40 ligation activates murine macrophages via an IFN-gamma-dependent mechanism resulting in tumor cell destruction in vitro. AB - We have shown previously that agonistic anti-CD40 mAb induced T cell-independent antitumor effects in vivo. In this study, we investigated mechanisms of macrophage activation with anti-CD40 mAb treatment, assessed by the antitumor action of macrophages in vitro. Intraperitoneal injection of anti-CD40 mAb into C57BL/6 mice resulted in activation of peritoneal macrophages capable of suppressing B16 melanoma cell proliferation in vitro, an effect that was greatly enhanced by LPS and observed against several murine and human tumor cell lines. Anti-CD40 mAb also primed macrophages in vitro to mediate cytostatic effects in the presence of LPS. The tumoristatic effect of CD40 ligation-activated macrophages was associated with apoptosis and killing of tumor cells. Activation of macrophages by anti-CD40 mAb required endogenous IFN-gamma because priming of macrophages by anti-CD40 mAb was abrogated in the presence of anti-IFN-gamma mAb, as well as in IFN-gamma-knockout mice. Macrophages obtained either from C57BL/6 mice depleted of T and NK cells by Ab treatment, or from scid/beige mice, were still activated by anti-CD40 mAb to mediate cytostatic activity. These results argued against the role of NK and T cells as the sole source of exogenous IFN gamma for macrophage activation and suggested that anti-CD40 mAb-activated macrophages could produce IFN-gamma. We confirmed this hypothesis by detecting intracytoplasmic IFN-gamma in macrophages activated with anti-CD40 mAb in vivo or in vitro. IFN-gamma production by macrophages was dependent on IL-12. Taken together, the results show that murine macrophages are activated directly by anti CD40 mAb to secrete IFN-gamma and mediate tumor cell destruction. PMID- 15879095 TI - Estrogen regulates CCR gene expression and function in T lymphocytes. AB - Estrogen has been implicated in the observed female bias in autoimmune diseases. However, the mechanisms behind this gender dimorphism are poorly defined. We have previously reported that in vivo T cell trafficking is gender- and estrogen dependent. Chemokine receptors are critical determinants of T cell homing and immune response. In this study, we show that the female gender is associated with increased CD4(+) T cell CCR1-CCR5 gene and protein expression in mice. The increased CCR expression correlates with enhanced in vitro chemotaxis response to MIP-1beta (CCL4). In vivo treatment of young oophorectomized and postmenopausal female mice with 17beta-estradiol also increased CD4(+) T cell CCR expression. Finally, 17beta-estradiol enhances tyrosine phosphorylation in T cells stimulated with MIP-1alpha in a time-dependent manner. Our results indicate an important role of estrogen in determining T cell chemokine response that may help explain the increased susceptibility and severity of autoimmune diseases in females. PMID- 15879096 TI - Cyclophilin A-deficient mice are resistant to immunosuppression by cyclosporine. AB - Cyclosporine is an immunosuppressive drug that is widely used to prevent organ transplant rejection. Known intracellular ligands for cyclosporine include the cyclophilins, a large family of phylogenetically conserved proteins that potentially regulate protein folding in cells. Immunosuppression by cyclosporine is thought to result from the formation of a drug-cyclophilin complex that binds to and inhibits calcineurin, a serine/threonine phosphatase that is activated by TCR engagement. Amino acids within the cyclophilins that are critical for binding to cyclosporine have been identified. Most of these residues are highly conserved within the 15 mammalian cyclophilins, suggesting that many are potential targets for the drug. We examined the effects of cyclosporine on immune cells and mice lacking Ppia, the gene encoding the prototypical cyclophilin protein cyclophilin A. TCR-induced proliferation and signal transduction by Ppia(-/-) CD4(+) T cells were resistant to cyclosporine, an effect that was attributable to diminished calcineurin inhibition. Immunosuppressive doses of cyclosporine failed to block the responses of Ppia(-/-) mice to allogeneic challenge. Rag2(-/-) mice reconstituted with Ppia(-/-) splenocytes were also cyclosporine resistant, indicating that this property is intrinsic to Ppia(-/-) immune cells. Thus, among multiple potential ligands, CypA is the primary mediator of immunosuppression by cyclosporine. PMID- 15879097 TI - Spontaneous and homeostatic proliferation of CD4 T cells are regulated by different mechanisms. AB - Transfer of naive CD4 T cells into lymphopenic mice initiates a proliferative response of the transferred cells, often referred to as homeostatic proliferation. Careful analysis reveals that some of the transferred cells proliferate rapidly and undergo robust differentiation to memory cells, a process we have designated spontaneous proliferation, and other cells proliferate relatively slowly and show more limited evidence of differentiation. In this study we report that spontaneous proliferation is IL-7 independent, whereas the slow proliferation (referred to as homeostatic proliferation) is IL-7 dependent. Administration of IL-7 induces homeostatic proliferation of naive CD4 T cells even within wild-type recipients. Moreover, the activation/differentiation pattern of the two responses are clearly distinguishable, indicating that different activation mechanisms may be involved. Our results reveal the complexity and heterogeneity of lymphopenia-driven T cell proliferation and suggest that they may have fundamentally distinct roles in the maintenance of CD4 T cell homeostasis. PMID- 15879098 TI - Gamma delta T cells respond directly to pathogen-associated molecular patterns. AB - Gammadelta T cells recognize unprocessed or non-peptide Ags, respond rapidly to infection, and localize to mucosal surfaces. We have hypothesized that the innate functions of gammadelta T cells may be more similar to those of cells of the myeloid lineage than to other T cells. To begin to test this assumption, we have analyzed the direct response of cultured human and peripheral blood bovine gammadelta T cells to pathogen associated molecular patterns (PAMPs) in the absence of APCs using microarray, real-time RT-PCR, proteome array, and chemotaxis assays. Our results indicate that purified gammadelta T cells respond directly to PAMPs by increasing expression of chemokine and activation-related genes. The response was distinct from that to known gammadelta T cell Ags and different from the response of myeloid cells to PAMPs. In addition, we have analyzed the expression of a variety of PAMP receptors in gammadelta T cells. Freshly purified bovine gammadelta T cells responded more robustly to PAMPs than did cultured human cells and expressed measurable mRNA encoding a variety of PAMP receptors. Our results suggest that rapid response to PAMPs through the expression of PAMP receptors may be another innate role of gammadelta T cells. PMID- 15879099 TI - Pertussis toxin (PTX) B subunit and the nontoxic PTX mutant PT9K/129G inhibit Tat induced TGF-beta production by NK cells and TGF-beta-mediated NK cell apoptosis. AB - We show that the pertussis toxin B oligomer (PTX-B), and the PTX mutant PT9K/129G, which is safely administered in vivo, inhibit both transcription and secretion of TGF-beta elicited by HIV-1 Tat in NK cells. Tat-induced TGF-beta mRNA synthesis is also blocked by the ERK1 inhibitor PD98059, suggesting that ERK1 is needed for TGF-beta production. Moreover, Tat strongly activates the c Jun component of the multimolecular complex AP-1, whereas TGF-beta triggers c-Fos and c-Jun. Of note, treatment of NK cells with PTX-B or PT9K/129G inhibits Tat- and TGF-beta-induced activation of AP-1. TGF-beta enhances starvation-induced NK cell apoptosis, significantly reduces transcription of the antiapoptotic protein Bcl-2, and inhibits Akt phosphorylation induced by oligomerization of the triggering NK cell receptor NKG2D. All these TGF-beta-mediated effects are prevented by PTX-B or PT9K/129G through a PI3K-dependent mechanism, as demonstrated by use of the specific PI3K inhibitor, LY294002. Finally, PTX-B and PT9K/129G up-regulate Bcl-x(L), the isoform of Bcl-x that protects cells from starvation-induced apoptosis. It is of note that in NK cells from patients with early HIV-1 infection, mRNA expression of Bcl-2 and Bcl-x(L) was consistently lower than that in healthy donors; interestingly, TGF-beta and Tat were detected in the sera of these patients. Together, these data suggest that Tat-induced TGF beta production and the consequent NK cell failure, possibly occurring during early HIV-1 infection, may be regulated by PTX-B and PT9K/129G. PMID- 15879100 TI - B cell receptor (BCR) cross-talk: CD40 engagement creates an alternate pathway for BCR signaling that activates I kappa B kinase/I kappa B alpha/NF-kappa B without the need for PI3K and phospholipase C gamma. AB - BCR signaling is propagated by a series of intermediaries and eventuates in NF kappaB activation, among other outcomes. Interruption of several mediators that constitute the signalosome, such as PI3K and phospholipase Cgamma2, completely blocks BCR signaling for NF-kappaB. We show here that this accepted, conventional paradigm is, in fact, limited to naive B cells. CD40L treatment reprograms normal B cells such that a novel, alternate pathway for BCR signaling is created. Through this alternate pathway BCR triggering induces nuclear NF-kappaB without the need for PI3K or for phospholipase Cgamma2. Induction of NF-kappaB via the alternate pathway is accompanied by IkappaB kinase beta (IKKbeta) phosphorylation, IkappaBalpha phosphorylation, and IkappaBalpha degradation, and inhibition of IKKbeta blocked IkappaBalpha degradation. Several key events in the conventional pathway, including early protein tyrosine phosphorylation, were unimpeded by generation of the alternate pathway which appears to operate in parallel, rather than in competition, with classical BCR signaling. These results demonstrate cross-talk between CD40 and BCR, such that the requirements for BCR signaling are altered by prior B cell exposure to CD40L. The alternate BCR signaling pathway bypasses multiple signalosome elements and terminates in IKKbeta activation. PMID- 15879101 TI - Selective expression of the 21-kilodalton tyrosine-phosphorylated form of TCR zeta promotes the emergence of T cells with autoreactive potential. AB - T cells undergo negative selection in the thymus to eliminate potentially autoreactive cells. The signals generated through the alphabeta TCR following receptor interactions with peptide/MHC complexes in the thymus control these selection processes. Following receptor ligation, a fraction of the TCR zeta subunit appears as two distinct tyrosine-phosphorylated forms of 21 and 23 kDa (p21 and p23). Previous data have reported elevated levels of p21 in some murine models of autoimmunity. We have examined the contributions of both the p21 and p23 to T cell negative selection in the HY TCR-transgenic system using ITAM substituted TCR zeta and CD3 epsilon transgenic mice. Expression of just p21, in the absence of p23, partially impairs negative selection of self-reactive HY specific T cells. This results in the emergence of potentially autoreactive peripheral T cells and an elevated population of CD11b(+)B220(+) B cells in the spleen. These data clearly identify a specific and unique role for p21 during negative selection. PMID- 15879102 TI - Preferential cell death of CD8+ effector memory (CCR7-CD45RA-) T cells by hydrogen peroxide-induced oxidative stress. AB - T cells are used in many cell-based cancer treatments. However, oxidative stress that is induced during various chronic inflammatory conditions, such as cancer, can impair the immune system and have detrimental effects on T cell function. In this study, we have investigated the sensitivity of different human T cell subsets to H(2)O(2)-induced oxidative stress. We showed that central memory (CD45RA(-)CCR7(+)) and effector memory (CD45RA(-)CCR7(-)) T cells are more sensitive to H(2)O(2) as compared with naive (CD45RA(+)CCR7(+)) T cells. Furthermore, the study showed that CD8(+) effector memory T cells are more sensitive to low levels of H(2)O(2) (5 microM) compared with other types of T cells investigated. H(2)O(2)-exposed CD45RO(+) T cells showed mitochondrial depolarization prior to caspase 3 activity. Moreover, the pan-caspase inhibitor z Val-Ala-Asp(OMe)-fluoromethylketone rescued cells from death. These experiments suggest that H(2)O(2)-induced cell death of CD45RO(+) T cells acts via the mitochondrial pathway and that caspase involvement is needed. This study suggests that oxidative stress in cancer patients can be disadvantageous for T cell-based adoptive cell transfer therapies, since effector memory T cells are the primary phenotype of the cells administered. PMID- 15879103 TI - Expression of killer cell lectin-like receptor G1 on antigen-specific human CD8+ T lymphocytes during active, latent, and resolved infection and its relation with CD57. AB - Killer cell lectin-like receptor G1 (KLRG1) is one of several inhibitory killer cell lectin-like receptors expressed by NK cells and T lymphocytes, mainly CD8(+) effector/memory cells that can secrete cytokines but have poor proliferative capacity. Using multiparameter flow cytometry, we studied KLRG1 expression on CD8(+) T cells specific for epitopes of CMV, EBV, influenza, and HIV. Over 92% of CD8(+) cells specific for CMV or EBV expressed KLRG1 during the latent stage of these chronic infections. CD8(+) T cell cells specific for HIV epitopes were mostly (72-89%) KLRG1(+), even though not quite at the level of predominance noted with CMV or EBV. Lower frequency of KLRG1 expression was observed among CD8(+) cells specific for influenza (40-73%), a resolved infection without a latent stage. We further observed that CD8(+) cells expressing CD57, a marker of replicative senescence, also expressed KLRG1; however, a population of CD57( )KLRG1(+) cells was also identified. This population may represent a "memory" phenotype, because they also expressed CD27, CD28, CCR7, and CD127. In contrast, CD57(+)KLRG1(+) cells did not express CD27, CD28, and CCR7, and expressed CD127 at a much lower frequency, indicating that they represent effector cells that are truly terminally differentiated. The combination of KLRG1 and CD57 expression might thus aid in refining functional characterization of CD8(+) T cell subsets. PMID- 15879104 TI - Reactive oxygen species and 12/15-lipoxygenase contribute to the antiproliferative capacity of alternatively activated myeloid cells elicited during helminth infection. AB - Understanding the role of CD11b(+)GR-1(+) myeloid suppressor cells in the immune suppression and immunoregulation associated with a variety of diseases may provide therapeutic opportunities. In this article, we show, in a model of helminth infection, that CD11b(+)GR-1(+) myeloid suppressor cells but not CD11b(+)F4/80(high) mature macrophages expanded in the peritoneal cavity of BALB/c mice implanted with Taenia crassiceps. Peritoneal cell populations from early stage-infected animals impaired T cell proliferation by secreting NO. Yet, they lost their ability to secrete NO in the late stage of infection. Concomitantly, their capacity to exert arginase activity and to express mRNAs coding for FIZZ1 (found in inflammatory zone 1), Ym, and macrophage galactose type C-type lectin increased. Furthermore, cells from early stage-infected mice triggered T cells to secrete IFN-gamma and IL-4, whereas in the late stage of infection, they only induced IL-4 production. These data suggest that CD11b(+)GR 1(+) myeloid suppressor cells displaying an alternative activation phenotype emerged gradually as T. crassiceps infection progressed. Corroborating the alternative activation status in the late stage of infection, the suppressive activity relied on arginase activity, which facilitated the production of reactive oxygen species including H(2)O(2) and superoxide. We also document that the suppressive activity of alternative myeloid suppressor cells depended on 12/15-lipoxygenase activation generating lipid mediators, which triggered peroxisome proliferator-activated receptor-gamma. IL-4 and IL-13 signaling contributed to the expansion of myeloid suppressor cells in the peritoneal cavity of T. crassiceps-infected animals and to their antiproliferative activity by allowing arginase and 12/15-lipoxygenase gene expression. PMID- 15879105 TI - Tid1 is required for T cell transition from double-negative 3 to double-positive stages. AB - Tid1, a DnaJ cochaperone protein, is the mammalian homologue of the Drosophila tumor suppressor Tid56 whose antitumor function is most likely mediated through its capacity to regulate cell differentiation in imaginal discs. We suspected that the mammalian counterpart, tid1, may also be involved in regulating cell differentiation. To investigate this, we exploited the system of T cell development to examine whether tid1 plays a role in this well-defined process. Mice with tid1 specifically deleted in T cells developed thymic atrophy, with dramatic reduction of double-positive and single-positive thymocytes in the tid1( /-) thymus. Although the subpopulations of tid1(-/-) double-negative (DN) 1-3 thymocytes were normal, the subpopulation of DN4 thymocytes was measurably smaller because of reduced proliferation and significant cell death. Immature tid1(-/-) thymocytes show normal VDJ beta-chain rearrangement and pre-TCR and CD3 expression in both DN3 and DN4 thymocytes, but in DN4 thymocytes, there was significantly reduced expression of the antiapoptotic bcl-2 gene. Restoring the expression level of Bcl-2 protein in tid1(-/-) thymus by introduction of a transgenic human bcl-2 gene resulted in reversal of the developmental defects in tid1(-/-) thymus. Together, these results demonstrate that tid1 is critical in early thymocyte development, especially during transition from the DN3 to double positive stages, possibly through its regulation of bcl-2 expression, which provides survival signals. PMID- 15879106 TI - CpG-independent synergistic induction of beta-chemokines and a dendritic cell phenotype by orthophosphorothioate oligodeoxynucleotides and granulocyte macrophage colony-stimulating factor in elutriated human primary monocytes. AB - Chemokines attract leukocytes bearing the relevant chemokine receptors and regulate innate immune responses. CpG oligodeoxynucleotides (ODN) and GM-CSF are potent vaccine adjuvants and in combination induce enhanced Th1 responses by mechanisms yet to be determined. We have examined combinations of CpG- or non-CpG ODN and GM-CSF for effects on the production of chemokines and the differentiation of monocytes to dendritic cells. High levels of the Th1 attracting, HIV-1-inhibitory chemokines, CCL3/MIP-1alpha and CCL4/MIP-1beta, were induced in human primary monocytes when CpG- or non-CpG-ODN was combined with GM CSF, but not with IL-4 or IFN-gamma. The synergistic induction of beta-chemokines by non-CpG-ODN was phosphorothioate (PS) chemistry dependent and inhibited by blocking endosome maturation/acidification and ERK1/2 activation. Chemokine and TLR9 mRNAs were induced by PS-ODN. Cells treated with non-CpG PS-ODN and GM-CSF expressed dendritic cell marker CD83 and high levels of HLA-DR and costimulatory molecules, and were CD14(-) or CD14(dim), consistent with monocyte differentiation into a dendritic cell phenotype. The induction of CD83 and beta chemokines was tyrosine phosphorylation dependent. Secreted CCL3 and CCL4 were detected as a heterodimer. Our results indicate the CpG-independent synergy between PS-ODN and GM-CSF mediated through chemokine and dendritic cell induction. In addition, our observations suggest that PS-ODN plus GM-CSF may be useful as potent ex vivo dendritic cell differentiation/maturation agents for dendritic cell therapy and as vaccine adjuvants for tumor and infectious microorganisms, including HIV-1. PMID- 15879107 TI - Impaired TGF-beta responses in peripheral T cells of G alpha i2-/- mice. AB - Null mutation of heterotrimeric G protein alpha2 inhibitory subunit (Galphai2) induces Th1-skewed hyperimmune responses in the colon, leading to chronic colitis and the development of colonic adenocarcinoma. However, the underlying molecular mechanisms and cellular basis, in particular, for the role of Galphai2 in regulating immune responses, are poorly understood. We show here that peripheral T cells from Galphai2-deficient mice do not respond normally to the inhibitory effects of TGF-beta on proliferation and cytokine production, revealing a previously unappreciated cross-talk between these two signaling pathways. Lack of Galphai2 resulted in decreased phosphorylation of Smad2 and Smad3 in T cells at the basal levels as well as at the late but not early phase of TGF-beta stimulation, which appears to be ascribed to differential expression of neither cell surface TGF-beta receptors nor Smad7. The altered phosphorylation of Smad proteins involves phospholipase C-mediated signaling, a downstream signaling molecule of Galphai2, because phospholipase C inhibitors could restore Smad2 and Smad3 phosphorylation in Galphai2(-/-) T cells at levels comparable to that in wild-type T cells. Moreover, adoptive transfer of Galphai2-deficient T cells into immunocompromised mice rendered an otherwise resistant mouse strain susceptible to trinitrobenzesulfonic acid-induced colitis, suggesting that an impaired response of Galphai2-deficient T cells to TGF-beta may be one of the primary defects accounting for the observed colonic Th1-skewed hyperimmune responses. These findings shed new lights on the molecular and cellular basis of how Galphai2 down-regulates immune responses, contributing to the maintenance of mucosal tolerance. PMID- 15879108 TI - Endosomal translocation of vertebrate DNA activates dendritic cells via TLR9 dependent and -independent pathways. AB - TLRs discriminate foreign from self via their specificity for pathogen-derived invariant ligands, an example being TLR9 recognizing bacterial unmethylated CpG motifs. In this study we report that endosomal translocation of CpG DNA via the natural endocytotic pathway is inefficient and highly saturable, whereas endosomal translocation of DNA complexed to the cationic lipid N-[1-(2,3 dioleoyloxy)propyl]-N,N,N-trimethylammonium methylsulfate (DOTAP) is not. Interestingly, DOTAP-mediated enhanced endosomal translocation of otherwise nonstimulatory vertebrate DNA or of certain noncanonical CpG motifs triggers robust dendritic cell activation in terms of both up-regulation of CD40/CD69 and cytokine production, such as type I IFN and IL-6. We report that the stimulatory activity of phosphorothioated noncanonical CpG oligodeoxynucleotides is TLR9 dependent, whereas phosphodiester DNA, such as vertebrate DNA, in addition trigger TLR9-independent pathways. We propose that the inefficiency of the natural route for DNA internalization hinders low affinity TLR9 ligands in endosomes to reach threshold concentrations required for TLR9 activation. Endosomal compartmentalization of TLR9 may thus reflect an evolutionary strategy to avoid TLR9 activation by self-DNA. PMID- 15879109 TI - TGF-alpha regulates TLR expression and function on epidermal keratinocytes. AB - The expression of TLRs on epithelial cells provides a first line of defense against invading pathogens. We investigated the regulated expression and function of TLR5 and TLR9 on human keratinocytes, because we found by immunohistochemistry that these TLRs are expressed in distinct layers of the epidermis. We found that TGF-alpha, a growth and differentiation factor that is present during wound healing and in psoriasis, increased the expression of both TLR5 and TLR9 on keratinocytes. In addition, TGF-alpha regulated the function of TLR5 and TLR9, because activation with their respective ligands enhanced the production of IL-8 and human beta-defensins. These findings provide evidence that TGF-alpha up regulates TLR expression and function, augmenting host defense mechanisms at epithelial surfaces. PMID- 15879110 TI - Transcriptional profiling of gamma delta T cells identifies a role for vitamin D in the immunoregulation of the V gamma 9V delta 2 response to phosphate containing ligands. AB - Vitamin D is a steroid hormone that, in addition to its well-characterized role in calcium/phosphate metabolism, has been found to have regulatory properties for immune system function. The nuclear vitamin D receptor is widely expressed in tissues, but has also been shown to be regulated by hormones, growth factors, and cytokines. In this study we show that activation of human Vdelta2Vgamma9 T cells by nonpeptidic monoalkyl phosphates such as isopentenyl pyrophosphate leads to the up-regulation of the vitamin D receptor via a pathway that involves the classical isoforms of protein kinase C. We further show that this receptor is active by demonstrating that the ligand 1alpha,25-dihydroxyvitamin D3 (vitD3) significantly inhibits in a dose-dependent fashion phospholigand-induced gammadelta T cell expansion, IFN-gamma production, and CD25 expression. We also show that vitD3 negatively regulates signaling via Akt and ERK and, at high concentrations, potentiates Ag-induced cell death. As such, these data provide further support for the immunoregulatory properties of vitamin D, and suggest that the ability of vitD3 to negatively regulate the proinflammatory activity of gammadelta T cells may contribute to the protection this vitamin affords against inflammatory and autoimmune disorders dependent upon Th1-type responses. PMID- 15879111 TI - An ancient role for a prokineticin domain in invertebrate hematopoiesis. AB - Hemopoietic development requires firm control of cell proliferation and differentiation. Although recent research has revealed conserved function of transcription factors and signaling pathways regulating lineage commitment in hemopoietic development in Drosophila melanogaster and vertebrates, little is known about hemopoietic cytokines among the invertebrate phyla. In the present study, we show that differentiation and growth of hemopoietic stem cells in vitro from an invertebrate, Pacifastacus leniusculus, require an endogenous cytokine like factor, astakine, containing a prokineticin (PK) domain. Astakine induces a strong hematopoiesis response in live animals. An astakine homologue was also found in the shrimp, Penaeus monodon. So far, PK domains are only identified in vertebrates, in which they, for example, direct angiogenic growth. Our finding of the first PK-like cytokine characterized from any invertebrate provides novel information concerning the evolution of growth factors and blood cell development. PMID- 15879112 TI - Testicular immune privilege promotes transplantation tolerance by altering the balance between memory and regulatory T cells. AB - Immune responses are suppressed in immunologically privileged sites, which may provide a unique opportunity to prolong allograft survival. However, it is unknown whether testicular immune privilege promotes transplantation tolerance. Mechanisms underlying immune privilege are also not well understood. Here we found that islet transplantation in the testis, an immunologically privileged site, generates much less memory CD8(+) T cells but induces more Ag-specific CD4(+)CD25(+) regulatory T cells than in a conventional site. These CD4(+)CD25(+) cells exhibited the suppression of alloimmune responses in vivo and in vitro. Despite the immune regulation, intratesticular islet allografts all were rejected within 42 days after transplantation although they survived longer than renal subcapsular islet allografts. However, blocking CD40/CD40L costimulation induced the tolerance of intratesticular, but not renal subcapsular, islet allografts. Tolerance to intratesticular islet allografts spread to skin allografts in the non-privileged sites. Either transfer of memory CD8(+) T cells or deletion of CD25(+) T cells in vivo broke islet allograft tolerance. Thus, transplantation tolerance requires both costimulatory blockade, which suppresses acute allograft rejection, and a favorable balance between memory and regulatory T cells that could favorably prevent late allograft failure. These findings reveal novel mechanisms of immune privilege and provide direct evidence that testicular immune privilege fosters the induction of transplantation tolerance to allografts in both immunologically privileged and non-privileged sites. PMID- 15879113 TI - TGF-beta inhibits Fas-mediated apoptosis of a follicular dendritic cell line by down-regulating the expression of Fas and caspase-8: counteracting role of TGF beta on TNF sensitization of Fas-mediated apoptosis. AB - Follicular dendritic cells (FDC) constitute the framework of germinal center (GC) in secondary lymphoid follicles, and the integrity of FDC networks is critically affected by cytokines present in the GC. We have previously shown that TNF promotes Fas-mediated apoptosis of HK cells, an established FDC-like cell line, by up-regulating Fas expression. However, in the developing GC, FDC death is not a hallmark of GC despite the presence of TNF and FasL. In this study, we report that TGF-beta inhibits Fas-mediated apoptosis of HK cells by down-regulating the expression of surface Fas and caspase-8. The inhibitory effect of TGF-beta can be observed when HK cells were simultaneously treated with TNF and TGF-beta, indicating that TGF-beta counteracts the effect of TNF in sensitizing cells to Fas-mediated apoptosis. Furthermore, the deprivation of TGF-beta by injecting neutralizing TGF-beta Abs to the SRBC-immunized mice resulted in the sporadic appearance of FDC undergoing apoptosis in the lymphoid follicles, suggesting that TGF-beta functions as a naturally occurring inhibitor that rescues FDCs which are predisposed to apoptosis. Our study documents a novel function of TGF-beta in the maintenance of FDC networks. PMID- 15879114 TI - Interallelic class switch recombination contributes significantly to class switching in mouse B cells. AB - Except for the expression of IgM and IgD, DNA recombination is constantly needed for the expression of other Ig classes and subclasses. The predominant path of class switch recombination (CSR) is intrachromosomal, and the looping-out and deletion model has been abundantly documented. However, switch regions also occasionally constitute convenient substrates for interchromosomal recombination, since it is noticeably the case in a number of chromosomal translocations causing oncogene deregulation in the course of lymphoma and myeloma. Although asymmetric accessibility of Ig alleles should theoretically limit its occurrence, interallelic CSR was shown to occur at low levels during IgA switching in rabbit, where the definition of allotypes within both V and C regions helped identify interchromosomally derived Ig. Thus, we wished to evaluate precisely interallelic CSR frequency in mouse B cells, by using a system in which only one allele (of b allotype) could express a functional VDJ region, whereas only interallelic CSR could restore expression of an excluded (a allotype) allele. In our study, we show that interchromosomal recombination of V(H) and Cgamma or Calpha occurs in vivo in B cells at a frequency that makes a significant contribution to physiological class switching: trans-association of V(H) and C(H) genes accounted for 7% of all alpha mRNA, and this frequency was about twice higher for the gamma3 transcripts, despite the much shorter distance between the J(H) region and the Cgamma3 gene, thus confirming that this phenomenon corresponded to site specific switching and not to random recombination between long homologous loci. PMID- 15879115 TI - Enforced expression of Spi-B reverses T lineage commitment and blocks beta selection. AB - The molecular changes that restrict multipotent murine thymocytes to the T cell lineage and render them responsive to Ag receptor signals remain poorly understood. In this study, we report our analysis of the role of the Ets transcription factor, Spi-B, in this process. Spi-B expression is acutely induced coincident with T cell lineage commitment at the CD4(-)CD8(-)CD44(-)CD25(+) (DN3) stage of thymocyte development and is then down-regulated as thymocytes respond to pre-TCR signals and develop beyond the beta-selection checkpoint to the CD4( )CD8(-)CD44(-)CD25(-) (DN4) stage. We found that dysregulation of Spi-B expression in DN3 thymocytes resulted in a dose-dependent perturbation of thymocyte development. Indeed, DN3 thymocytes expressing approximately five times the endogenous level of Spi-B were arrested at the beta-selection checkpoint, due to impaired induction of Egr proteins, which are important molecular effectors of the beta-selection checkpoint. T lineage-committed DN3 thymocytes expressing even higher levels of Spi-B were diverted to the dendritic cell lineage. Thus, we demonstrate that the prescribed modulation of Spi-B expression is important for T lineage commitment and differentiation beyond the beta-selection checkpoint; and we provide insight into the mechanism underlying perturbation of development when that expression pattern is disrupted. PMID- 15879116 TI - Mechanisms of dimethyl sulfoxide augmentation of IL-1 beta production. AB - Expression of the inflammatory cytokine IL-1beta occurs in various inflammatory diseases, and IL-1beta production is regulated at multiple levels. There are conflicting reports about the effects of antioxidants on IL-1beta production. In this study, we investigated the regulatory role of the antioxidant DMSO on LPS stimulated IL-1beta gene expression in human PBMC and in vivo. This study demonstrated that 1% DMSO increased LPS-stimulated (50 ng/ml) IL-1beta secretion in a dose- and time-dependent manner without altering TNF or IL-6. DMSO also elevated IL-1beta secretion by PBMC in response to exogenous superoxide anions. Despite the increase in IL-1beta, there was no augmentation of NF-kappaB with the addition of DMSO. The steady state mRNA coding for IL-1beta following LPS stimulation was also increased. Cycloheximide studies demonstrated that the DMSO augmentation of IL-1beta mRNA did not require de novo protein synthesis, and studies with actinomycin D showed that DMSO did not alter the half-life of IL 1beta mRNA, suggesting that DMSO did not change the stability of IL-1beta mRNA. Experiments using a reporter vector containing the 5'-flanking region of the human IL-1beta gene revealed that DMSO augmented LPS-induced IL-1beta reporter activity. In vivo, treatment of mice with DMSO significantly increased plasma levels of IL-1beta after endotoxin challenge. These data indicate that DMSO directly increases LPS-stimulated IL-1beta protein production through the mechanisms of augmenting promoter activity and increasing mRNA levels. PMID- 15879117 TI - IFN-gamma-stimulated transcriptional activation by IFN-gamma-activated transcriptional element-binding factor 1 occurs via an inducible interaction with CAAAT/enhancer-binding protein-beta. AB - IFN-gamma-activated transcriptional element (GATE)-binding factor 1 (GBF1) was identified as a transactivator that induces gene expression through GATE, a novel IFN-inducible element. Although it can induce gene expression, it is an extremely weak DNA-binding protein on its own. GATE also binds another transcription factor, C/EBP-beta. Therefore, we explored whether GBF1 physically interacts with C/EBP-beta to induce IFN-gamma-regulated transcription. In response to IFN-gamma, C/EBP-beta undergoes phosphorylation at a critical ERK1/2 phosphorylation motif. Mutational inactivation of this motif and/or interference with the ERK1/2 activation prevented the IFN-gamma-induced interactions between GBF1 and C/EBP beta. A 37-aa long peptide derived from the GBF1 protein can associate with C/EBP beta in an IFN-inducible manner. These results identify a converging point for two transactivators that exert their effects through a single response element. Together, our studies identify a novel regulatory mechanism that controls IFN induced transcription. PMID- 15879118 TI - Demethylation of the same promoter sequence increases CD70 expression in lupus T cells and T cells treated with lupus-inducing drugs. AB - Exposing genetically predisposed individuals to certain environmental agents is believed to cause human lupus. How environmental agents interact with the host to cause lupus is poorly understood. Procainamide and hydralazine are drugs that cause lupus in genetically predisposed individuals. Understanding how these environmental agents cause lupus may indicate mechanisms relevant to the idiopathic disease. Abnormal T cell DNA methylation, a repressive epigenetic DNA modification, is implicated in procainamide and hydralazine induced lupus, as well as idiopathic lupus. Procainamide is a competitive DNA methyltransferase (Dnmt) inhibitor, hydralazine inhibits ERK pathway signaling thereby decreasing Dnmt expression, and in lupus T cells decreased ERK pathway signaling causing a similar Dnmt decrease. T cells treated with procainamide, hydralazine, and other Dnmt and ERK pathway inhibitors cause lupus in mice. Whether the same genetic regulatory elements demethylate in T cells treated with Dnmt inhibitors, ERK pathway inhibitors, and in human lupus is unknown. CD70 (TNFSF7) is a B cell costimulatory molecule overexpressed on CD4(+) lupus T cells as well as procainamide and hydralazine treated T cells, and contributes to excessive B cell stimulation in vitro and in lupus. In this report we identify a genetic element that suppresses CD70 expression when methylated, and which demethylates in lupus and in T cells treated with Dnmt and ERK pathway inhibitors including procainamide and hydralazine. The results support a model in which demethylation of specific genetic elements in T cells, caused by decreasing Dnmt expression or inhibiting its function, contributes to drug-induced and idiopathic lupus through altered gene expression. PMID- 15879119 TI - Beyond the 12/23 rule of VDJ recombination independent of the Rag proteins. AB - The combinatorial repertoire of AgRs is established through somatic recombination of V, D, and J gene segments during lymphocyte development. Incorporation of D segments into IgH, TCRbeta, and TCRdelta chains also contributes to junctional diversification by substantially extending the length of the third CDR. The V, D, and J gene segments are flanked by recombination signals (RS) of 12- or 23-mer spacer length that direct recombination according to the 12/23 rule. D genes in the TCRbeta and TCRdelta loci are flanked by a 12RS and 23RS, and their incorporation is controlled by mechanisms "beyond the 12/23 rule." In the TCRbeta locus, selective interactions between Rag proteins and the RS flanking the V-D and D-J genes, respectively, are sufficient to enforce D gene usage. In this article, we report that in the TCRdelta locus, the Rag proteins are not the major determinant of D gene incorporation. In developing mouse and human thymocytes, the two Ddelta genes rearrange predominantly to form D-D coding joints. In contrast, when tested in ex vivo transfection assays in a nonlymphoid cell line, the flanking RS mediate deletion, rather than incorporation, of the two D genes on both exogenous recombination substrates and the endogenous locus. These results suggest that selective Rag-RS interactions are not the sole regulators of D gene segment incorporation, and additional, perhaps lymphocyte-specific, mechanisms exist that allow proper shaping of the primary AgR repertoire. PMID- 15879120 TI - Recombinant C345C and factor I modules of complement components C5 and C7 inhibit C7 incorporation into the complement membrane attack complex. AB - Complement component C5 binds to components C6 and C7 in reversible reactions that are distinct from the essentially nonreversible associations that form during assembly of the complement membrane attack complex (MAC). We previously reported that the approximately 150-aa residue C345C domain (also known as NTR) of C5 mediates these reversible reactions, and that the corresponding recombinant module (rC5-C345C) binds directly to the tandem pair of approximately 75-residue factor I modules from C7 (C7-FIMs). We suggested from these and other observations that binding of the C345C module of C5 to the FIMs of C7, but not C6, is also essential for MAC assembly itself. The present report describes a novel method for assembling a complex that appears to closely resemble the MAC on the sensor chip of a surface plasmon resonance instrument using the complement reactive lysis mechanism. This method provides the ability to monitor individually the incorporation of C7, C8, and C9 into the complex. Using this method, we found that C7 binds to surface-bound C5b,6 with a K(d) of approximately 3 pM, and that micromolar concentrations of either rC5-C345C or rC7 FIMs inhibit this early step in MAC formation. We also found that similar concentrations of either module inhibited complement-mediated erythrocyte lysis by both the reactive lysis and classical pathway mechanisms. These results demonstrate that the interaction between the C345C domain of C5 and the FIMs of C7, which mediates reversible binding of C5 to C7 in solution, also plays an essential role in MAC formation and complement lytic activity. PMID- 15879121 TI - A new tyrosine phosphorylation site in PLC gamma 1: the role of tyrosine 775 in immune receptor signaling. AB - Phospholipase Cgamma (PLCgamma) is a ubiquitous gatekeeper of calcium mobilization and diacylglycerol-mediated events induced by the activation of Ag and growth factor receptors. The activity of PLCgamma is regulated through its controlled membrane translocation and tyrosine (Y) phosphorylation. Four activation-induced tyrosine phosphorylation sites have been previously described (Y472, Y771, Y783, and Y1254), but their specific roles in Ag receptor-induced PLCgamma1 activation are not fully elucidated. Unexpectedly, we found that the phosphorylation of a PLCgamma1 construct with all four sites mutated to phenylalanine was comparable with that observed with wild-type PLCgamma1, suggesting the existence of an unidentified site(s). Sequence alignment with known phosphorylation sites in PLCgamma2 indicated homology of PLCgamma1 tyrosine residue 775 (Y775) with PLCgamma2 Y753, a characterized phosphorylation site. Tyrosine 775 was characterized as a phosphorylation site using phospho-specific anti-Y775 antiserum, and by mutational analysis. Phosphorylation of Y775 did not depend on the other tyrosines, and point mutation of PLCgamma1 Y775, or the previously described Y783, substantially reduced AgR-induced calcium, NF-AT, and AP-1 activation. Mutation of Y472, Y771, and Y1254 had no effect on overall PLCgamma1 phosphorylation or activation. Although the concomitant mutation of Y775 and Y783 abolished downstream PLCgamma1 signaling, these two tyrosines were sufficient to reconstitute the wild-type response in the absence of functional Y472, Y771, and Y1254. These data establish Y775 as a critical phosphorylation site for PLCgamma1 activation and confirm the functional importance of Y783. PMID- 15879122 TI - Transcriptional regulation of the human CD3 gamma gene: the TATA-less CD3 gamma promoter functions via an initiator and contiguous Sp-binding elements. AB - Growing evidence that the CD3gamma gene is specifically targeted in some T cell diseases focused our attention on the need to identify and characterize the elusive elements involved in CD3gamma transcriptional control. In this study, we show that while the human CD3gamma and CD3delta genes are oriented head-to-head and separated by only 1.6 kb, the CD3gamma gene is transcribed from an independent but weak, lymphoid-specific TATA-less proximal promoter. Using RNA ligase-mediated rapid amplification of cDNA ends, we demonstrate that a cluster of transcription initiation sites is present in the vicinity of the primary core promoter, and the major start site is situated in a classical initiator sequence. A GT box immediately upstream of the initiator binds Sp family proteins and the general transcription machinery, with the activity of these adjacent elements enhanced by the presence of a second GC box 10 nt further upstream. The primary core promoter is limited to a sequence that extends upstream to -15 and contains the initiator and GT box. An identical GT box located approximately 50 nt from the initiator functions as a weak secondary core promoter and likely generates transcripts originating upstream from the +1. Finally, we show that two previously identified NFAT motifs in the proximal promoter positively (NFATgamma(1)) or negatively (NFATgamma(1) and NFATgamma(2)) regulate expression of the human CD3gamma gene by their differential binding of NFATc1 plus NF-kappaB p50 or NFATc2 containing complexes, respectively. These data elucidate some of the mechanisms controlling expression of the CD3gamma gene as a step toward furthering our understanding of how its transcription is targeted in human disease. PMID- 15879123 TI - Human factor H-related protein 5 has cofactor activity, inhibits C3 convertase activity, binds heparin and C-reactive protein, and associates with lipoprotein. AB - Factor H-related protein 5 (FHR-5) is a recently discovered member of the factor H (fH)-related protein family. FHR proteins are structurally similar to the complement regulator fH, but their biological functions remain poorly defined. FHR-5 is synthesized in the liver and consists of 9 short consensus repeats (SCRs), which display various degrees of homology to those of fH and the other FHR proteins. FHR-5 colocalizes with complement deposits in vivo and binds C3b in vitro, suggesting a role in complement regulation or localization. The current study examined whether rFHR-5 exhibits properties similar to those of fH, including heparin binding, CRP binding, cofactor activity for the factor I mediated degradation of C3b and decay acceleration of the C3 convertase. rFHR-5 bound heparin-BSA and heparin-agarose and a defined series of truncations expressed in Pichia pastoris localized the heparin-binding region to within SCRs 5-7. rFHR-5 bound CRP, and this binding was also localized to SCRs 5-7. FHR-5 inhibited alternative pathway C3 convertase activity in a fluid phase assay; however, dissociation of the convertase was not observed in a solid phase assay. rFHR-5 displayed factor I-dependent cofactor activity for C3b cleavage, although it was apparently less effective than fH. In addition, we demonstrate association of FHR-5 with high density lipid lipoprotein complexes in human plasma. These results demonstrate that FHR-5 shares properties of heparin and CRP binding and lipoprotein association with one or more of the other FHRs but is unique among this family of proteins in possessing independent complement-regulatory activity. PMID- 15879124 TI - Mouse cathelin-related antimicrobial peptide chemoattracts leukocytes using formyl peptide receptor-like 1/mouse formyl peptide receptor-like 2 as the receptor and acts as an immune adjuvant. AB - Mammalian antimicrobial proteins, such as defensins and cathelicidin, have stimulating effects on host leukocytes. Cathelin-related antimicrobial peptide (CRAMP), the orthologue of human cathelicidin/LL-37, is the sole identified murine cathelicidin. CRAMP has been shown to have both antimicrobial and angiogenic activities. However, whether CRAMP, like human cathelicidin/LL-37, also exhibits a direct effect on the migration and function of leukocytes is not known. We have observed that CRAMP, like LL-37, was chemotactic for human monocytes, neutrophils, macrophages, and mouse peripheral blood leukocytes. CRAMP also induced calcium mobilization and the activation of MAPK in monocytes. CRAMP induced calcium flux in monocytes was desensitized by MMK-1, an agonistic ligand specific for formyl peptide receptor-like-1 (FPRL1), and vice versa, suggesting the use of FPRL1 by CRAMP as a receptor. Furthermore, CRAMP induced the chemotaxis of human embryonic kidney 293 cells transfected with either FPRL1 or mouse formyl peptide receptor-2, the mouse homologue of FPRL1, but not by untransfected parental human embryonic kidney 293 cells, confirming the use of FPRL1/mouse formyl peptide receptor-2 by CRAMP. Injection of CRAMP into mouse air pouches resulted in the recruitment predominantly of neutrophils and monocytes, indicating that CRAMP acts as a chemotactic factor in vivo. Finally, simultaneous administration of OVA with CRAMP to mice promoted both humoral and cellular Ag specific immune responses. Thus, CRAMP functions as both a chemoattractant for phagocytic leukocytes and an enhancer of adaptive immune response. PMID- 15879125 TI - Protective immunity to genital herpes simplex [correction of simpex] virus type 2 infection is mediated by T-bet. AB - We show, for the first time, that the transcription factor T-bet, which is implicated in IFN-gamma production, is required for the induction of vaccine induced antiviral immune protection. T-bet was found to be important in both the innate and acquired immune protection against genital HSV-2 infection. T-bet(-/-) and T-bet(+/+) mice were infected vaginally with HSV-2 and examined daily for disease and mortality. T-bet(-/-) mice had significantly higher virus titers than T-bet(+/+) mice following a primary HSV-2 infection, and succumbed significantly earlier to the infection. This result was associated with an impaired NK cell cytotoxic capacity and NK cell-mediated IFN-gamma production in the T-bet(-/-) mice. To assess the induction of acquired antiviral immune protection, mice were vaccinated with an attenuated virus before infection. Vaccinated T-bet(-/-) mice could not control viral replication following an HSV-2 challenge and had significantly higher virus titers and mortality rates than vaccinated T-bet(+/+) mice that remained healthy. The impaired acquired immune protection in T-bet(-/-) mice was associated with a significantly decreased HSV-2-specific delayed-type hypersensitivity response and a significantly reduced HSV-2-specific IFN-gamma production from CD4(+) T cells. However, T-bet deficiency did not impair either the IFN-gamma production or the cytotoxic capacity of HSV-2-specific CD8(+) T cells. We conclude that T-bet plays a crucial role in both the innate defense and the generation of vaccine-induced immunity against genital HSV-2 infection in mice. PMID- 15879126 TI - Analysis of the 5q31-q33 locus shows an association between IL13-1055C/T IL-13 591A/G polymorphisms and Schistosoma haematobium infections. AB - Millions of humans are exposed to schistosome infections, which cause severe kidney and liver disease and 280,000 deaths annually. Th2-mediated immunity is critical to human defenses against this pathogen and susceptibility to infection is controlled by a major genetic locus that includes IL4, IL5, and IL13 genes. These observations led us to evaluate whether certain polymorphisms in IL4, IL5, or IL13 determine schistosome infection. The study was performed in two Dogon villages where Schistosoma haematobium is endemic. Schistosome infections were evaluated by counting eggs and measuring worm Ags in urine. Genetic polymorphisms were determined by restriction enzyme analysis or by primer extension and denaturing high-performance liquid chromatography analysis. Associations were tested using family-based association tests and logistical regression analysis. The alleles IL13-1055C (p = 0.05) and IL13-591A (p = 0.01) are shown, by family based association test, to be preferentially transmitted to children with the 10% highest infections. A logistic regression analysis that included IL13-1055 G/G, G/T and T/T genotypes, age, gender, and village of residency, applied to the whole study population, showed that subjects bearing the IL13-1055T/T genotype were on average much less infected than individuals with other genotypes. Previous studies on asthma indicated that the IL13-1055T allele increased gene transcription, which is in agreement with the fact that this cytokine enhances resistance to infection by schistosome in humans. PMID- 15879127 TI - Dendritic cells cross-present exogenous fungal antigens to stimulate a protective CD8 T cell response in infection by Histoplasma capsulatum. AB - The contribution of CD8 T cells in host defense against histoplasmosis is minor in the CD4 T cell-intact mouse, as it has been shown that depleting CD8 T cells only marginally affects fungal clearance. However, it remains to be determined whether the CD8 T cells are protective in a host lacking functional CD4 T cells. In this study, MHC class II-deficient mice infected with Histoplasma capsulatum (Histoplasma) kept the fungus in check for up to 16 wk, indicating that CD8 T cells are able to limit fungal replication. Ex vivo studies showed that CD8 T cells from Histoplasma-infected mice expressed both intracytoplasmic IFN-gamma and granzyme B. Furthermore, CD8 T cells exhibited cytotoxic activity against macrophage targets containing Histoplasma. We demonstrated that the macrophage, being the primary host cell as well as the effector cell, can also serve as Ag donor to dendritic cells. Histoplasma-specific CD8 T cells are stimulated by dendritic cells that present exogenous Histoplasma Ags, either through direct ingestion of yeasts or through uptake of apoptotic macrophage-associated fungal Ags, a process known as "cross-presentation." Based on these results, we present a model detailing the possible sequence of events leading to a cell-mediated immune response and fungal clearance in Histoplasma-infected hosts. PMID- 15879128 TI - Electroporation as a "prime/boost" strategy for naked DNA vaccination against a tumor antigen. AB - We have developed novel DNA fusion vaccines encoding tumor Ags fused to pathogen derived sequences. This strategy activates linked T cell help and, using fragment C of tetanus toxin, amplification of anti-tumor Ab, CD4(+), and CD8(+) T cell responses is achievable in mice. However, there is concern that simple DNA vaccine injection may produce inadequate responses in larger humans. To overcome this, we tested electroporation as a method to increase the transfection efficiency and immune responses by these tumor vaccines in vivo in mice. Using a DNA vaccine expressing the CTL epitope AH1 from colon carcinoma CT26, we confirmed that effective priming and tumor protection in mice are highly dependent on vaccine dose and volume. However, suboptimal vaccination was rendered effective by electroporation, priming higher levels of AH1-specific CD8(+) T cells able to protect mice from tumor growth. Electroporation during priming with our optimal vaccination protocol did not improve CD8(+) T cell responses. In contrast, electroporation during boosting strikingly improved vaccine performance. The prime/boost strategy was also effective if electroporation was used at both priming and boosting. For Ab induction, DNA vaccination is generally less effective than protein. However, prime/boost with naked DNA followed by electroporation dramatically increased Ab levels. Thus, the priming qualities of DNA fusion vaccines, integrated with the improved Ag expression offered by electroporation, can be combined in a novel homologous prime/boost approach, to generate superior antitumor immune responses. Therefore, boosting may not require viral vectors, but simply a physical change in delivery, facilitating application to the cancer clinic. PMID- 15879129 TI - Binding of the complement inhibitor C4bp to serogroup B Neisseria meningitidis. AB - Neisseria meningitidis (meningococcus) is an important cause of meningitis and sepsis. Currently, there is no effective vaccine against serogroup B meningococcal infection. Host defense against neisseriae requires the complement system (C) as indicated by the fact that individuals deficient in properdin or late C components (C6-9) have an increased susceptibility to recurrent neisserial infections. Because the classical pathway (CP) is required to initiate efficient complement activation on neisseriae, meningococci should be able to evade it to cause disease. To test this hypothesis, we studied the interactions of meningococci with the major CP inhibitor C4b-binding protein (C4bp). We tested C4bp binding to wild-type group B meningococcus strain (H44/76) and to 11 isogenic mutants thereof that differed in capsule expression, lipo oligosaccharide sialylation, and/or expression of either porin (Por) A or PorB3. All strains expressing PorA bound radiolabeled C4bp, whereas the strains lacking PorA bound significantly less C4bp. Increased binding was observed under hypotonic conditions. Deleting PorB3 did not influence C4bp binding, but the presence of polysialic acid capsule reduced C4bp binding by 50%. Bound C4bp remained functionally active in that it promoted the inactivation of C4b by factor I. PorA-expressing strains were also more resistant to C lysis than PorA negative strains in a serum bactericidal assay. Binding of C4bp thus helps Neisseria meningitidis to escape CP complement activation. PMID- 15879130 TI - The mucosal adjuvant macrophage-activating lipopeptide-2 directly stimulates B lymphocytes via the TLR2 without the need of accessory cells. AB - The macrophage-activating lipopeptide-2 (MALP-2) is an agonist of the TLR heterodimer 2/6, which exhibits potent activity as mucosal adjuvant, promoting strong humoral and cellular responses. Although B cells expressing TLR2/6 are potential targets, very little is known about the effect of MALP-2 on B cells. Studies were performed using total spleen cells or purified B cells from WT mice or animals deficient in TLR2, T cells, B cells, or specific subpopulations of B cells. They demonstrated that MALP-2 promotes a T cell-independent activation and maturation of B cells (mainly follicular but also B-1a and marginal zone B cells) via TLR2. MALP-2 also increased the frequency of IgM- and IgG-secreting cells, but bystander cells were required for IgA secretion. Activated B cells exhibited increased expression of activation markers and ligands that are critical for cross-talk with T cells (CD19, CD25, CD80, CD86, MHC I, MHC II, and CD40). Immunization of mice lacking T cells showed that MALP-2-mediated stimulation of TLR2/6 was unable to circumvent the need of T cell help for efficient Ag-specific B cell activation. Immunization of mice lacking B cells demonstrated that B cells are critical for MALP-2-dependent improvement of T cell responses. The knowledge emerging from this work suggests that MALP-2-mediated activation of B cells through TLR2/6 is critical for adjuvanticity. B cell stimulation by pattern recognition receptors seems to be a basic mechanism that can be exploited to improve the immunogenicity of vaccine formulations. PMID- 15879131 TI - Caspase-8 activity prevents type 2 cytokine responses and is required for protective T cell-mediated immunity against Trypanosoma cruzi infection. AB - During Trypanosoma cruzi infection, T cells up-regulate caspase-8 activity. To assess the role of caspase-8 in T cell-mediated immunity, we investigated the effects of caspase-8 inhibition on T cells in viral FLIP (v-FLIP) transgenic mice. Compared with wild-type controls, increased parasitemia was observed in v FLIP mice infected with T. cruzi. There was a profound decrease in expansion of both CD4 and CD8 T cell subsets in the spleens of infected v-FLIP mice. We did not find differences in activation ratios of T cells from transgenic or wild-type infected mice. However, the numbers of memory/activated CD4 and CD8 T cells were markedly reduced in v-FLIP mice, possibly due to defective survival. We also found decreased production of IL-2 and increased secretion of type 2 cytokines, IL-4 and IL-10, which could enhance susceptibility to infection. Similar, but less pronounced, alterations were observed in mice treated with the caspase-8 inhibitor, zIETD. Furthermore, blockade of caspase-8 by zIETD in vitro mimicked the effects observed on T. cruzi infection in vivo, affecting the generation of activated/memory T cells and T cell cytokine production. Caspase-8 is also required for NF-kappaB signaling upon T cell activation. Blockade of caspase-8 by either v-FLIP expression or treatment with zIETD peptide decreased NF-kappaB responses to TCR:CD3 engagement in T cell cultures. These results suggest a critical role for caspase-8 in the establishment of T cell memory, cell signaling, and regulation of cytokine responses during protozoan infection. PMID- 15879132 TI - Protein kinase R mediates intestinal epithelial gene remodeling in response to double-stranded RNA and live rotavirus. AB - As sentinels of host defense, intestinal epithelial cells respond to the viral pathogen rotavirus by activating a gene expression that promotes immune cell recruitment and activation. We hypothesized that epithelial sensing of rotavirus might target dsRNA, which can be detected by TLR3 or protein kinase R (PKR). Accordingly, we observed that synthetic dsRNA, polyinosinic acid:cytidylic acid (poly(I:C)), potently induced gene remodeling in model intestinal epithelia with the specific pattern of expressed genes, including both classic proinflammatory genes (e.g., IL-8), as well as genes that are classically activated in virus infected cells (e.g., IFN-responsive genes). Poly(I:C)-induced IL-8 was concentration dependent (2-100 mug/ml) and displayed slower kinetics compared with IL-8 induced by bacterial flagellin (ET(50) approximately 24 vs 8 h poly(I:C) vs flagellin, respectively). Although model epithelia expressed detectable TLR3 mRNA, neither TLR3-neutralizing Abs nor chloroquine, which blocks activation of intracellular TLR3, attenuated epithelial responses to poly(I:C). Conversely, poly(I:C)-induced phosphorylation of PKR and inhibitors of PKR, 2 aminopurine and adenine, ablated poly(I:C)-induced gene expression but had no effect on gene expression induced by flagellin, thus suggesting that intestinal epithelial cell detection of dsRNA relies on PKR. Consistent with poly(I:C) detection by an intracellular molecule such as PKR, we observed that both uptake of and responses to poly(I:C) were polarized to the basolateral side. Lastly, we observed that the pattern of pharmacologic inhibition of responses to poly(I:C) was identical to that seen in response to infection by live rotavirus, indicating a potentially important role for PKR in activating intestinal epithelial gene expression in rotavirus infection. PMID- 15879133 TI - Exchanging ESAT6 with TB10.4 in an Ag85B fusion molecule-based tuberculosis subunit vaccine: efficient protection and ESAT6-based sensitive monitoring of vaccine efficacy. AB - Previously we have shown that Ag85B-ESAT-6 is a highly efficient vaccine against tuberculosis. However, because the ESAT-6 Ag is also an extremely valuable diagnostic reagent, finding a vaccine as effective as Ag85B-ESAT-6 that does not contain ESAT-6 is a high priority. Recently, we identified a novel protein expressed by Mycobacterium tuberculosis designated TB10.4. In most infected humans, TB10.4 is strongly recognized, raising interest in TB10.4 as a potential vaccine candidate and substitute for ESAT-6. We have now examined the vaccine potential of this protein and found that vaccination with TB10.4 induced a significant protection against tuberculosis. Fusing Ag85B to TB10.4 produced an even more effective vaccine, which induced protection against tuberculosis comparable to bacillus Calmette-Guerin vaccination and superior to the individual Ag components. Thus, Ag85B-TB10 represents a new promising vaccine candidate against tuberculosis. Furthermore, having now exchanged ESAT-6 for TB10.4, we show that ESAT-6, apart from being an excellent diagnostic reagent, can also be used as a reagent for monitoring vaccine efficacy. This may open a new way for monitoring vaccine efficacy in clinical trials. PMID- 15879134 TI - Control of infection with Leishmania major in susceptible BALB/c mice lacking the common gamma-chain for FcR is associated with reduced production of IL-10 and TGF beta by parasitized cells. AB - Previous studies have shown that the in vitro ligation of FcgammaRs with IgG opsonized Leishmania amastigotes promotes IL-10 production by macrophages. In addition, infection of either BALB/c mice lacking the common gamma-chain of Fc receptors (FcgammaR(-/-)) or mice genetically altered to lack circulating Ab (J(H)D) with Leishmania pifanoi results in reduced and delayed lesion development and a deficit in the recruitment of inflammatory cells into infected lesions. We show in this study that FcgammaR(-/-) mice can control infection with Leishmania major and totally resolve cutaneous lesions. The ability to eventually control infection is not associated with a reduction in lesion inflammation or a reduction in the ability of Leishmania to parasitize cells through week 6 of infection. The immune response in healing FcgammaR(-/-) mice is associated with a reduction in numbers of cells producing Th2-type cytokines, including IL-4 and IL 10, but not an increase in numbers of IFN-gamma-producing cells characteristic of a dominant Th1-type response. Instead, we observe a reduction in levels of IL-10 and TGF-beta within infected lesions, including reduced levels of these cytokines within parasitized macrophages. Together, these results suggest that uptake of opsonized parasites via FcgammaRs may be a strong in vivo stimulus for the production of anti-inflammatory cytokines that play a role in susceptibility to infection. PMID- 15879135 TI - Role of IFN-gamma in regulating T2 immunity and the development of alternatively activated macrophages during allergic bronchopulmonary mycosis. AB - Pulmonary Cryptococcus neoformans infection of C57BL/6 mice is an established model of a chronic pulmonary fungal infection accompanied by an "allergic" response (T2) to the infection, i.e., a model of an allergic bronchopulmonary mycosis. Our objective was to determine whether IFN-gamma plays a role in regulating the pulmonary T2 immune response in C. neoformans-infected C57BL/6 mice. Long-term pulmonary fungistasis was lost in IFN-gamma knockout (KO) mice, resulting in an increased pulmonary burden of fungi at wk 3. IFN-gamma was required for the early influx of leukocytes into the lungs but was not required later in the infection. By wk 3, eosinophil and macrophage numbers were elevated in the absence of IFN-gamma. The inducible NO synthase to arginase ratio was lower in the lungs of IFN-gamma KO mice and the macrophages had increased numbers of intracellular cryptococci and YM1 crystals, indicative of alternatively activated macrophages in these mice. There was evidence of pulmonary fibrosis in both wild-type and IFN-gamma KO mice by 5 wk postinfection. IFN-gamma production was not required for the development of T2 cytokine (IL-4, IL-5, IL-13) producing cells in the lungs and lung-associated lymph nodes or induction of an IgE response. At a number of time points, T2 cytokine production was enhanced in IFN gamma KO mice. Thus, in the absence of IFN-gamma, C57BL/6 mice develop an augmented allergic response to C. neoformans, including enhanced generation of alternatively activated macrophages, which is accompanied by a switch from a chronic to a progressive pulmonary cryptococcal infection. PMID- 15879136 TI - Cyclic AMP response element-binding protein positively regulates production of IFN-gamma by T cells in response to a microbial pathogen. AB - IFN-gamma is essential for resistance to many intracellular pathogens, including Mycobacterium tuberculosis. Transcription of the IFN-gamma gene in activated T cells is controlled by the proximal promoter element (-73 to -48 bp). CREB binds to the IFN-gamma proximal promoter, and binding is enhanced by phosphorylation of CREB. Studies in human T cell lines and in transgenic mice have yielded conflicting results about whether CREB is a positive or a negative regulator of IFN-gamma transcription. To determine the role of CREB in mediating IFN-gamma production in response to a microbial pathogen, we evaluated the peripheral blood T cell response to M. tuberculosis in healthy tuberculin reactors. EMSAs, chromatin immunoprecipitation, and Western blotting demonstrated that stimulation of PBMC with M. tuberculosis induced phosphorylation and enhanced binding of CREB to the IFN-gamma proximal promoter. Neutralization of CREB with intracellular Abs or down-regulation of CREB levels with small interfering RNA decreased M. tuberculosis-induced production of IFN-gamma and IFN-gamma mRNA expression. In addition, M. tuberculosis-stimulated T cells from tuberculosis patients, who have ineffective immunity, showed diminished IFN-gamma production, reduced amounts of CREB binding to the IFN-gamma proximal promoter, and absence of phosphorylated CREB. These findings demonstrate that CREB positively regulates IFN-gamma production by human T cells that respond to M. tuberculosis. PMID- 15879137 TI - Insights into pathogen immune evasion mechanisms: Anaplasma phagocytophilum fails to induce an apoptosis differentiation program in human neutrophils. AB - Polymorphonuclear leukocytes (PMNs or neutrophils) are essential to human innate host defense. However, some bacterial pathogens circumvent destruction by PMNs and thereby cause disease. Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, survives within PMNs in part by altering normal host cell processes, such as production of reactive oxygen species (ROS) and apoptosis. To investigate the molecular basis of A. phagocytophilum survival within neutrophils, we used Affymetrix microarrays to measure global changes in human PMN gene expression following infection with A. phagocytophilum. Notably, A. phagocytophilum uptake induced fewer perturbations in host cell gene regulation compared with phagocytosis of Staphylococcus aureus. Although ingestion of A. phagocytophilum did not elicit significant PMN ROS, proinflammatory genes were gradually up-regulated, indicating delayed PMN activation rather than loss of proinflammatory capacity normally observed during phagocytosis-induced apoptosis. Importantly, ingestion of A. phagocytophilum failed to trigger the neutrophil apoptosis differentiation program that typically follows phagocytosis and ROS production. Heat-killed A. phagocytophilum caused some similar initial alterations in neutrophil gene expression and function, which included delaying normal PMN apoptosis and blocking Fas-induced programmed cell death. However, at 24 h, down-regulation of PMN gene transcription may be more reliant on active infection. Taken together, these findings suggest two separate antiapoptotic processes may work concomitantly to promote bacterial survival: 1) uptake of A. phagocytophilum fails to trigger the apoptosis differentiation program usually induced by bacteria, and 2) a protein or molecule on the pathogen surface can mediate an early delay in spontaneous neutrophil apoptosis. PMID- 15879138 TI - Gastrointestinal ischemia-reperfusion injury is lectin complement pathway dependent without involving C1q. AB - Complement activation plays an important role in local and remote tissue injury associated with gastrointestinal ischemia-reperfusion (GI/R). The role of the classical and lectin complement pathways in GI/R injury was evaluated using C1q deficient (C1q KO), MBL-A/C-deficient (MBL-null), complement factor 2- and factor B-deficient (C2/fB KO), and wild-type (WT) mice. Gastrointestinal ischemia (20 min), followed by 3-h reperfusion, induced intestinal and lung injury in C1q KO and WT mice, but not in C2/fB KO mice. Addition of human C2 to C2/fB KO mice significantly restored GI/R injury, demonstrating that GI/R injury is mediated via the lectin and/or classical pathway. Tissue C3 deposition in C1q KO and WT, but not C2/fB KO, mice after GI/R demonstrated that complement was activated in C1q KO mice. GI/R significantly increased serum alanine aminotransferase, gastrointestinal barrier dysfunction, and neutrophil infiltration into the lung and gut in C1q KO and WT, but not C2/fB KO, mice. MBL-null mice displayed little gut injury after GI/R, but lung injury was present. Addition of recombinant human MBL (rhuMBL) to MBL-null mice significantly increased injury compared with MBL null mice after GI/R and was reversed by anti-MBL mAb treatment. However, MBL null mice were not protected from secondary lung injury after GI/R. These data demonstrate that C2 and MBL, but not C1q, are necessary for gut injury after GI/R. Lung injury in mice after GI/R is MBL and C1q independent, but C2 dependent, suggesting a potential role for ficolins in this model. PMID- 15879139 TI - Proteinase-3 induces procaspase-3 activation in the absence of apoptosis: potential role of this compartmentalized activation of membrane-associated procaspase-3 in neutrophils. AB - In the present study, we provide evidence that procaspase-3 is a novel target of proteinase 3 (PR3) but not of human neutrophil elastase (HNE). Human mast cell clone 1 (HMC1) and rat basophilic leukemia (RBL) mast cell lines were transfected with PR3 or the inactive mutated PR3 (PR3S203A) or HNE cDNA. In both RBL/PR3 and HMC1/PR3, a constitutive activity of caspase-3 was measured with DEVD substrate, due to the direct processing of procaspase-3 by PR3. No caspase-3 activation was observed in cells transfected with the inactive PR3 mutant or HNE. Despite the high caspase-3 activity in RBL/PR3, no apoptosis was detected as demonstrated by an absence of 1) phosphatidylserine externalization, 2) mitochondria cytochrome c release, 3) upstream caspase-8 or caspase-9 activation, or 4) DNA fragmentation. In vitro, purified PR3 cleaved procaspase-3 into an active 22-kDa fragment. In neutrophils, the 22-kDa caspase-3 activation fragment was present only in resting neutrophils but was absent after apoptosis. The 22 kDa fragment was specific of myeloid cells because it was absent from resting lymphocytes. This 22-kDa fragment was not present when neutrophils were treated with pefabloc, an inhibitor of serine proteinase. Like in HMC1/PR3, the 22-kDa caspase-3 fragment was restricted to the plasma membrane compartment. Double immunofluorescence labeling after streptolysin-O permeabilization further showed that PR3 and procaspase-3 could colocalize in an extragranular compartment. In conclusion, our results strongly suggest that compartmentalized PR3-induced caspase-3 activation might play specific functions in neutrophil survival. PMID- 15879140 TI - Estradiol is required for a proper immune response to bacterial and viral pathogens in the female brain. AB - Although the neuroprotective effects of estrogens are well recognized, the exact mechanisms involved in the ability of these sex steroids to protect the cerebral tissue still remain unclear. We tested in our study the hypothesis that estradiol (E(2)) modulates the innate immune response and expression of genes encoding proteins that a provide survival signal to neurons during infection. Mice received a single systemic or cerebral injection of LPS to trigger a robust but transient inflammatory reaction in the brain. The endotoxin increased transcriptional activation of genes encoding TLR2, TNF-alpha, and IL-12 in microglial cells. Expression of these transcripts was largely inhibited in the brain of ovariectomized mice at time 24 h postchallenge. E(2) replacement therapy totally rescued the ability of the endotoxin to trigger microglial cells and these permissive effects of E(2) are mediated via the estrogen receptor (ER)alpha. Indeed, ERalpha-deficient mice exhibited an inappropriate reaction to LPS when compared with ERbeta-deficient and wild-type mice. This defective innate immune response was also associated with a widespread viral replication and neurodegeneration in ovariectomized mice inoculated intranasally with HSV-2. These data provide evidence that interaction of E(2) with their nuclear ERalpha plays a critical role in the control of cytokines involved in the transfer from the innate to adaptive immunity. This transfer is deviant in mice lacking E(2), which allows pathogens to hide from immune surveillance and exacerbates neuronal damages during viral encephalitis. PMID- 15879141 TI - Neutrophil-derived heparin-binding protein (HBP/CAP37) deposited on endothelium enhances monocyte arrest under flow conditions. AB - In acute inflammation, infiltration of neutrophils often precedes a second phase of monocyte invasion, and data in the literature suggest that neutrophils may directly stimulate mobilization of monocytes via neutrophil granule proteins. In this study, we present a role for neutrophil-derived heparin-binding protein (HBP) in monocyte arrest on endothelium. Adhesion of neutrophils to bovine aorta endothelial cells (ECs) or HUVEC-triggered secretion of HBP and binding of the protein to the EC surface. Blockade of neutrophil adhesion by treatment with a mAb to CD18 greatly reduced accumulation of HBP. In a flow chamber model, immobilized recombinant HBP induced arrest of human monocytes or monocytic Mono Mac 6 (MM6) cells to activated EC or plates coated with recombinant adhesion molecules (E-selectin, P-selectin, VCAM-1). However, immobilized recombinant HBP did not influence arrest of neutrophils or lymphocytes. Treatment of MM6 cells with recombinant HBP evoked a rapid and clear-cut increase in cytosolic free Ca(2+) that was found to be critical for the HBP-induced monocyte arrest inasmuch as pretreatment with the intracellular calcium chelating agent BAPTA-AM abolished the evoked increase in adhesion. Thus, secretion of a neutrophil granule protein, accumulating on the EC surface and promoting arrest of monocytes, could contribute to the recruitment of monocytes at inflammatory loci. PMID- 15879142 TI - Junctional adhesion molecule-C regulates the early influx of leukocytes into tissues during inflammation. AB - Leukocyte recruitment from blood to inflammatory sites occurs in a multistep process that involves discrete molecular interactions between circulating and endothelial cells. Junctional adhesion molecule (JAM)-C is expressed at different levels on endothelial cells of lymphoid organs and peripheral tissues and has been proposed to regulate neutrophil migration by its interaction with the leukocyte integrin Mac-1. In the present study, we show that the accumulation of leukocytes in alveoli during acute pulmonary inflammation in mice is partially blocked using neutralizing Abs against JAM-C. To confirm the function of JAM-C in regulating leukocyte migration in vivo, we then generated a strain of transgenic mice overexpressing JAM-C under the control of the endothelial specific promotor Tie2. The transgenic animals accumulate more leukocytes to inflammatory sites compared with littermate control mice. Intravital microscopy shows that this is the result of increased leukocyte adhesion and transmigration, whereas rolling of leukocytes is not significantly affected in transgenic mice compared with littermates. Thus, JAM-C participates in the later steps of the leukoendothelial adhesion cascade. PMID- 15879143 TI - A synthetic TLR4 antagonist has anti-inflammatory effects in two murine models of inflammatory bowel disease. AB - Current evidence indicates that the chronic inflammation observed in the intestines of patients with inflammatory bowel disease is due to an aberrant immune response to enteric flora. We have developed a lipid A-mimetic, CRX-526, which has antagonistic activity for TLR4 and can block the interaction of LPS with the immune system. CRX-526 can prevent the expression of proinflammatory genes stimulated by LPS in vitro. This antagonist activity of CRX-526 is directly related to its structure, particularly secondary fatty acyl chain length. In vivo, CRX-526 treatment blocks the ability of LPS to induce TNF-alpha release. Importantly, treatment with CRX-526 inhibits the development of moderate-to severe disease in two mouse models of colonic inflammation: the dextran sodium sulfate model and multidrug resistance gene 1a-deficient mice. By blocking the interaction between enteric bacteria and the innate immune system, CRX-526 may be an effective therapeutic molecule for inflammatory bowel disease. PMID- 15879144 TI - Regulation of c-Jun phosphorylation by the I kappa B kinase-epsilon complex in fibroblast-like synoviocytes. AB - Rheumatoid arthritis (RA) causes a symmetric, inflammatory polyarthritis that results in joint destruction and significant disability. Signaling pathways that regulate the production of cytokines and destructive enzymes have been implicated in its pathogenesis and represent potential therapeutic targets. The IkappaB kinase (IKK)-related kinase, IKKepsilon/IKKi, which plays a pivotal role in regulating antiviral gene transcription, is constitutively expressed by cultured fibroblast-like synoviocytes (FLS) and could participate in the pathogenesis of RA. In the current studies we demonstrate that IKKepsilon protein is expressed in RA and osteoarthritis synovium and that the protein is found primarily in the synovial intimal lining. Functional studies in cultured FLS showed that IKKepsilon kinase activity is rapidly induced by cytokines, although IkappaB phosphorylation is significantly less compared with IKK2. Because NF-kappaB activation is similar in wild-type and IKKepsilon knockout murine FLS, studies were performed to identify an alternative substrate for IKKepsilon. Interestingly, c-Jun is a more efficient substrate for IKKepsilon immunocomplexes in human FLS and this activity appears to be independent of JNK. The functional relevance of IKKepsilon was examined using murine IKKepsilon(-/-) cultured FLS. IL-1-, TNF-alpha-, and LPS-mediated induction of matrix metalloproteinases, MMP3 and MMP13, is significantly decreased in the IKKepsilon(-/-) cells. These data suggest a novel role for the IKKepsilon complex in synovial inflammation, extracellular matrix destruction, and activation of the viral program and innate immune response in RA. PMID- 15879145 TI - Questioning current concepts in acute pancreatitis: endotoxin contamination of porcine pancreatic elastase is responsible for experimental pancreatitis associated distant organ failure. AB - The systemic inflammatory response syndrome is responsible for pancreatitis associated mortality. Recent in vitro and in vivo studies have suggested that pancreatic elastase is one missing link between the localized inflammatory process in the pancreas and distant organ dysfunction and failure. It has been shown that pancreatic elastase activates transcription factors, including NF kappaB, and induces TNF-alpha secretion in myeloid cells via TLRs. In this study we demonstrate that a highly purified low endotoxin pancreatic elastase preparation (El-UP) failed both to activate NF-kappaB and to induce TNF-alpha release in RAW 264.7 cells and bone marrow-derived macrophages. In contrast, a less purified elastase preparation (El-IV) caused activation of NF-kappaB and was able to induce TNF-alpha release at very low concentrations. These effects were sensitive to pretreatment of the cells with polymyxin B and were resistant to heat inactivation. Endotoxin activity as determined by the Limulus amebocyte lysate assay was >3 orders of magnitude lower in the low endotoxin elastase preparation (El-UP) compared with less purified elastase preparations (El-IV). In contrast to contaminated elastase or LPS, elastase free of contamination (El-UP) failed to induce elevated serum TNF-alpha levels or pulmonary neutrophil infiltration after i.p. application in mice and did not induce lethality when coinjected with d-galactosamine. Failure of low endotoxin elastase (El-UP) to induce proinflammatory effects in vivo and in vitro was not due to functional inactivity of the elastase preparation, as determined by elastase activity assay. These results question current concepts of direct proinflammatory effects attributed to pancreatic elastase. PMID- 15879146 TI - Exosomes derived from IL-10-treated dendritic cells can suppress inflammation and collagen-induced arthritis. AB - We have demonstrated previously that local, adenoviral-mediated gene transfer of viral IL-10 to a single joint of rabbits and mice with experimental arthritis can suppress disease in both the treated and untreated contralateral joints. This contralateral effect is mediated in part by APCs able to traffic from the treated joint to lymph nodes as well as to untreated joints. Moreover, injection of dendritic cells (DC) genetically modified to express IL-4 or Fas ligand was able to reverse established murine arthritis. To examine the ability of exosomes derived from immunosuppressive DCs to reduce inflammation and autoimmunity, murine models of delayed-type hypersensitivity and collagen-induced arthritis were used. In this study, we demonstrate that periarticular administration of exosomes purified from either bone marrow-derived DCs transduced ex vivo with an adenovirus expressing viral IL-10 or bone marrow-derived DCs treated with recombinant murine IL-10 were able to suppress delayed-type hypersensitivity responses within injected and untreated contralateral joints. In addition, the systemic injection of IL-10-treated DC-derived exosomes was able suppress the onset of murine collagen-induced arthritis as well as reduce severity of established arthritis. Taken together, these data suggest that immature DCs are able to secrete exosomes that are involved in the suppression of inflammatory and autoimmune responses. Thus DC-derived exosomes may represent a novel, cell-free therapy for the treatment of autoimmune diseases. PMID- 15879147 TI - Role of lipoteichoic acid in the phagocyte response to group B streptococcus. AB - Group B Streptococcus (GBS) cell walls potently activate phagocytes by a largely TLR2-independent mechanism. In contrast, the cell wall component lipoteichoic acid (LTA) from diverse Gram-positive bacterial species has been shown to engage TLR2. In this study we examined the role of LTA from GBS in phagocyte activation and the requirements for TLR-LTA interaction. Using cells from knockout mice and genetic complementation in epithelial cells we found that highly pure LTA from both GBS and Staphylococcus aureus interact with TLR2 and TLR6, but not TLR1, in contrast to previous reports. Furthermore, NF-kappaB activation by LTA required the integrity of two putative PI3K binding domains within TLR2 and was inhibited by wortmannin, indicating an essential role for PI3K in cellular activation by LTA. However, LTA from GBS proved to be a relatively weak stimulus of phagocytes containing approximately 20% of the activity observed with LTA from Staphylococcus aureus. Structural analysis by nuclear magnetic resonance spectrometry revealed important differences between LTA from GBS and S. aureus, specifically differences in glycosyl linkage, in the glycolipid anchor and a lack of N-acetylglucosamine substituents of the glycerophosphate backbone. Furthermore, GBS expressing LTA devoid of d-alanine residues, that are essential within immune activation by LTA, exhibited similar inflammatory potency as GBS with alanylated LTA. In conclusion, LTA from GBS is a TLR2/TLR6 ligand that might contribute to secreted GBS activity, but does not contribute significantly to GBS cell wall mediated macrophage activation. PMID- 15879148 TI - A critical role for sphingosine kinase in anaphylatoxin-induced neutropenia, peritonitis, and cytokine production in vivo. AB - The aim of our study was to investigate the roles played by sphingosine kinase (SPHK) in the anaphylatoxin C5a-triggered responses in vivo. Our data show that i.v. administration of C5a triggers a rapid neutropenic response, but pretreating mice with the SPHK inhibitor, N,N-dimethylsphingosine (DMS), 10 min before the C5a i.v. administration substantially inhibited the C5a-triggered neutropenia. Similarly the i.v. administration of C5a caused a rapid increase in the serum levels of TNF-alpha and IL-6, and this increase in cytokine levels was blocked by DMS. We then induced acute peritonitis with C5a. The C5a i.p. injection triggered a fast recruitment of neutrophils, later followed by monocytes, into the peritoneal cavity. Vascular permeability was also observed: when we i.v. injected Evans blue before C5a i.p. injection, we could observe a continued influx of the dye into the peritoneum. In mice pretreated with DMS, there was a significant reduction on the C5a-triggered neutrophil and monocyte infiltration, as well as a marked reduction on the Evans blue influx. Our data also show that the i.p. administration of C5a caused a rapid increase in TNF-alpha and IL-6 levels in the peritoneal cavity, and this increase in cytokine levels was substantially inhibited in mice pretreated with the SPHK inhibitor. Taken together, these observations suggest a potential role for SPHK in the C5a-triggered inflammatory responses in vivo. PMID- 15879149 TI - A direct role for C1 inhibitor in regulation of leukocyte adhesion. AB - Plasma C1 inhibitor (C1INH) is a natural inhibitor of complement and contact system proteases. Heterozygosity for C1INH deficiency results in hereditary angioedema, which is mediated by bradykinin. Treatment with plasma C1INH is effective not only in patients with hereditary angioedema, but also in a variety of other disease models, in which such therapy is accompanied by diminished neutrophil infiltration. The underlying mechanism has been explained primarily as a result of the inhibition of the complement and contact systems. We have shown that C1INH expresses the sialyl-Lewis(x) tetrasaccharide on its N-linked glycan, via which it binds to E- and P-selectins and interferes with leukocyte endothelial adhesion in vitro. Here we show that both native C1INH and reactive center cleaved C1INH significantly inhibit selectin-mediated leukocyte adhesion in several in vitro and in vivo models, whereas N-deglycosylated C1INH loses such activities. The data support the hypothesis that C1INH plays a direct role in leukocyte-endothelial cell adhesion, that the activity is mediated by carbohydrate, and that it is independent of protease inhibitory activity. Direct involvement of C1INH in modulation of selectin-mediated cell adhesion may be an important mechanism in the physiologic suppression of inflammation, and may partially explain its utility in therapy of inflammatory diseases. PMID- 15879150 TI - TLR4, but not TLR2, signals autoregulatory apoptosis of cultured microglia: a critical role of IFN-beta as a decision maker. AB - TLRs mediate diverse signaling after recognition of evolutionary conserved pathogen-associated molecular patterns such as LPS and lipopeptides. Both TLR2 and TLR4 are known to trigger a protective immune response as well as cellular apoptosis. In this study, we present evidence that TLR4, but not TLR2, mediates an autoregulatory apoptosis of activated microglia. Brain microglia underwent apoptosis upon stimulation with TLR4 ligand (LPS), but not TLR2 ligands (Pam(3)Cys-Ser-Lys(4), peptidoglycan, and lipoteichoic acid). Based on studies using TLR2-deficient or TLR4 mutant mice and TLR dominant-negative mutants, we also demonstrated that TLR4, but not TLR2, is necessary for microglial apoptosis. The critical difference between TLR2 and TLR4 signalings in microglia was IFN regulatory factor-3 (IRF-3) activation, followed by IFN-beta expression: while TLR4 agonist induced the activation of IRF-3/IFN-beta pathway, TLR2 did not. Nevertheless, both TLR2 and TLR4 agonists strongly induced NF-kappaB activation and NO production in microglia. Neutralizing Ab against IFN-beta attenuated TLR4 mediated microglial apoptosis. IFN-beta alone, however, did not induce a significant cell death. Meanwhile, TLR2 activation induced microglial apoptosis with help of IFN-beta, indicating that IFN-beta production following IRF-3 activation determines the apoptogenic action of TLR signaling. TLR4-mediated microglial apoptosis was mediated by MyD88 and Toll/IL-1R domain-containing adaptor-inducing IFN-beta, and was associated with caspase-11 and -3 activation rather than Fas-associated death domain protein/caspase-8 pathway. Taken together, TLR4 appears to signal a microglial apoptosis via autocrine/paracrine IFN-beta production, which may act as an apoptotic sensitizer. PMID- 15879151 TI - Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells. AB - Ethical considerations constrain the in vivo study of human hemopoietic stem cells (HSC). To overcome this limitation, small animal models of human HSC engraftment have been used. We report the development and characterization of a new genetic stock of IL-2R common gamma-chain deficient NOD/LtSz-scid (NOD-scid IL2Rgamma(null)) mice and document their ability to support human mobilized blood HSC engraftment and multilineage differentiation. NOD-scid IL2Rgamma(null) mice are deficient in mature lymphocytes and NK cells, survive beyond 16 mo of age, and even after sublethal irradiation resist lymphoma development. Engraftment of NOD-scid IL2Rgamma(null) mice with human HSC generate 6-fold higher percentages of human CD45(+) cells in host bone marrow than with similarly treated NOD-scid mice. These human cells include B cells, NK cells, myeloid cells, plasmacytoid dendritic cells, and HSC. Spleens from engrafted NOD-scid IL2Rgamma(null) mice contain human Ig(+) B cells and lower numbers of human CD3(+) T cells. Coadministration of human Fc-IL7 fusion protein results in high percentages of human CD4(+)CD8(+) thymocytes as well human CD4(+)CD8(-) and CD4(-)CD8(+) peripheral blood and splenic T cells. De novo human T cell development in NOD scid IL2Rgamma(null) mice was validated by 1) high levels of TCR excision circles, 2) complex TCRbeta repertoire diversity, and 3) proliferative responses to PHA and streptococcal superantigen, streptococcal pyrogenic exotoxin. Thus, NOD-scid IL2Rgamma(null) mice engrafted with human mobilized blood stem cells provide a new in vivo long-lived model of robust multilineage human HSC engraftment. PMID- 15879152 TI - Activation of discoidin domain receptor 1 on CD14-positive bronchoalveolar lavage fluid cells induces chemokine production in idiopathic pulmonary fibrosis. AB - Discoidin domain receptor 1 (DDR1) is a receptor tyrosine kinase activated by collagen. We previously reported the functional expression of DDR1 on human monocyte-derived macrophages in vitro; however, information regarding its role in diseases is limited. Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease, and the lesions contain an abundance of collagen. In this study, we examined DDR1 expression on bronchoalveolar lavage fluid (BALF) cells and investigated its functionality using samples obtained from 28 IPF patients, 13 chronic obstructive pulmonary disease patients, and 14 healthy volunteers. The DDR1 expression level in CD14-positive BALF cells was higher in IPF patients than in chronic obstructive pulmonary disease patients or healthy volunteers. The predominant isoform was DDR1b in the IPF group, while DDR1a was predominant in the other two groups. Using immunohistochemical analysis, we also detected DDR1 expression on infiltrating inflammatory cells in the IPF lesion. In IPF patients, DDR1 activation induced the production of MCP-1, IL-8, MIP-1 alpha, and matrix metalloproteinase-9 (MMP-9) from CD14-positive BALF cells in a p38 MAPK-dependent manner. In contrast, DDR1 activation of CD14-positive BALF cells in the other groups did not induce the production of these chemokines or MMP-9. These chemokines and MMP-9 contribute to the development of IPF and, therefore, we suggest that DDR1 might be associated with the pathogenesis of IPF in the tissue microenvironment. PMID- 15879153 TI - Absence of recipient CCR5 promotes early and increased allospecific antibody responses to cardiac allografts. AB - Acute rejection is mediated by T cell infiltration of allografts, but mechanisms mediating the delayed rejection of allografts in chemokine receptor-deficient recipients remain unclear. The rejection of vascularized, MHC-mismatched cardiac allografts by CCR5(-/-) recipients was investigated. Heart grafts from A/J (H 2(a)) donors were rejected by wild-type C57BL/6 (H-2(b)) recipients on day 8-10 posttransplant vs day 8-11 by CCR5(-/-) recipients. When compared with grafts from wild-type recipients, however, significant decreases in CD4(+) and CD8(+) T cells and macrophages were observed in rejecting allografts from CCR5-deficient recipients. These decreases were accompanied by significantly lower numbers of alloreactive T cells developing to IFN-gamma-, but not IL-4-producing cells in the CCR5(-/-) recipients, suggesting suboptimal priming of T cells in the knockout recipients. CCR5 was more prominently expressed on activated CD4(+) than CD8(+) T cells in the spleens of allograft wild-type recipients and on CD4(+) T cells infiltrating the cardiac allografts. Rejecting cardiac allografts from wild type recipients had low level deposition of C3d that was restricted to the graft vessels. Rejecting allografts from CCR5(-/-) recipients had intense C3d deposition in the vessels as well as on capillaries throughout the graft parenchyma similar to that observed during rejection in donor-sensitized recipients. Titers of donor-reactive Abs in the serum of CCR5(-/-) recipients were almost 20-fold higher than those induced in wild-type recipients, and the high titers appeared as early as day 6 posttransplant. These results suggest dysregulation of alloreactive Ab responses and Ab-mediated cardiac allograft rejection in the absence of recipient CCR5. PMID- 15879154 TI - Human 60-kDa heat shock protein is a target autoantigen of T cells derived from atherosclerotic plaques. AB - Epidemiological studies suggest the potential importance of an inflammatory component in atherosclerosis and support the hypothesis that immune responses to Ags of pathogens cross-react with homologous host proteins due to molecular mimicry. Protein candidates involved may be the stress-induced proteins known as heat shock proteins (HSP). In this study, we report that atherosclerotic plaques harbor in vivo-activated CD4(+) T cells that recognize the human 60-kDa HSP. Such in vivo-activated 60-kDa HSP-specific T cells are not detectable in the peripheral blood. In patients with positive serology and PCR for Chlamydia pneumoniae DNA, but not in patients negative for both, most of plaque-derived T cells specific for human 60-kDa HSP also recognized the C. pneumoniae 60-kDa HSP. We characterized the submolecular specificity of such 60-kDa HSP-specific plaque derived T cells and identified both the self- and cross-reactive epitopes of that autoantigen. On challenge with human 60-kDa HSP, most of the plaque-derived T cells expressed Th type 1 functions, including cytotoxicity and help for monocyte tissue factor production. We suggest that arterial endothelial cells, undergoing classical atherosclerosis risk factors and conditioned by Th type 1 cytokines, express self 60-kDa HSP, which becomes target for both autoreactive T cells and cross-reactive T cells to microbial 60-kDa HSP via a mechanism of molecular mimicry. This hypothesis is in agreement with the notion that immunization with HSP exacerbates atherosclerosis, whereas immunosuppression and T cell depletion prevent the formation of arteriosclerotic lesions in experimental animals. PMID- 15879155 TI - Nucleotide-binding oligomerization domain-2 modulates specific TLR pathways for the induction of cytokine release. AB - The recognition of peptidoglycan by cells of the innate immune system has been controversial; both TLR2 and nucleotide-binding oligomerization domain-2 (NOD2) have been implicated in this process. In the present study we demonstrate that although NOD2 is required for recognition of peptidoglycan, this leads to strong synergistic effects on TLR2-mediated production of both pro- and anti inflammatory cytokines. Defective IL-10 production in patients with Crohn's disease bearing loss of function mutations of NOD2 may lead to overwhelming inflammation due to a subsequent Th1 bias. In addition to the potentiation of TLR2 effects, NOD2 is a modulator of signals transmitted through TLR4 and TLR3, but not through TLR5, TLR9, or TLR7. Thus, interaction between NOD2 and specific TLR pathways may represent an important modulatory mechanism of innate immune responses. PMID- 15879156 TI - Gene transfer of tissue inhibitor of metalloproteinases-3 reverses the inhibitory effects of TNF-alpha on Fas-induced apoptosis in rheumatoid arthritis synovial fibroblasts. AB - Apart from counteracting matrix metalloproteinases, tissue inhibitor of metalloproteinases-3 (TIMP-3) has proapoptotic properties. These features have been attributed to the inhibition of metalloproteinases involved in the shedding of cell surface receptors such as the TNFR. However, little is known about effects of TIMP-3 in cells that are not susceptible to apoptosis by TNF-alpha. In this study, we report that gene transfer of TIMP-3 into human rheumatoid arthritis synovial fibroblasts and MRC-5 human fetal lung fibroblasts facilitates apoptosis and completely reverses the apoptosis-inhibiting effects of TNF-alpha. Although TNF-alpha inhibits Fas/CD95-induced apoptosis in untransfected and mock transfected cells, fibroblasts ectopically expressing TIMP-3 are sensitized most strongly to Fas/CD95-mediated cell death by TNF-alpha. Neither synthetic MMP inhibitors nor glycosylated bioactive TIMP-3 are able to achieve these effects. Gene transfer of TIMP-3 inhibits the TNF-alpha-induced activation of NF-kappaB in rheumatoid arthritis synovial fibroblasts and reduces the up-regulation of soluble Fas/CD95 by TNF-alpha, but has no effects on the cell surface expression of Fas. Collectively, our data demonstrate that intracellularly produced TIMP-3 not only induces apoptosis, but also modulates the apoptosis-inhibiting effects of TNF-alpha in human rheumatoid arthritis synovial fibroblast-like cells. Thus, our findings may stimulate further studies on the therapeutic potential of gene transfer strategies with TIMP-3. PMID- 15879157 TI - Hepatitis C virus drives the unconstrained monoclonal expansion of VH1-69 expressing memory B cells in type II cryoglobulinemia: a model of infection driven lymphomagenesis. AB - Chronic hepatitis C virus infection causes B cell lymphoproliferative disorders that include type II mixed cryoglobulinemia and lymphoma. This virus drives the monoclonal expansion and, occasionally, the malignant transformation of B cells producing a polyreactive natural Ab commonly encoded by the V(H)1-69 variable gene. Owing to their property of producing natural Ab, these cells are reminiscent of murine B-1 and marginal zone B cells. We used anti-Id Abs to track the stages of differentiation and clonal expansion of V(H)1-69(+) cells in patients with type II mixed cryoglobulinemia. By immunophenotyping and cell size analysis, we could define three discrete stages of differentiation of V(H)1-69(+) B cells: naive (small, IgM(high)IgD(high)CD38(+)CD27(-)CD21(high)CD95(-)CD5(-)), "early memory" (medium-sized, IgM(high)IgD(low)CD38( )CD27(+)CD21(low)CD95(+)CD5(+)), and "late memory" (large-sized, IgM(low)IgD(low neg)CD38(-)CD27(low)CD21(low-neg)CD5(-)CD95(-)). The B cells expanded in cryoglobulinemia patients have a "memory" phenotype; this fact, together with the evidence for intraclonal variation, suggests that antigenic stimulation by hepatitis C virus causes the unconstrained expansion of activated V(H)1-69(+) B cells. In some cases, these cells replace the entire pool of circulating B cells, although the absolute B cell number remains within normal limits. Absolute monoclonal V(H)1-69(+) B lymphocytosis was seen in three patients with cryoglobulinemia and splenic lymphoma; in two of these patients, expanded cells carried trisomy 3q. The data presented here indicate that the hepatitis C virus driven clonal expansion of memory B cells producing a V(H)1-69(+) natural Ab escapes control mechanisms and subverts B cell homeostasis. Genetic alterations may provide a further growth advantage leading to an overt lymphoproliferative disorder. PMID- 15879158 TI - Comparison of killer Ig-like receptor genotyping and phenotyping for selection of allogeneic blood stem cell donors. AB - The repertoire of killer Ig-like receptors (KIRs) can be determined at the level of DNA, RNA, or surface protein expression for selection of blood stem cell donors. We compared genotyping and phenotyping of the four inhibitory KIRs that are important in transplantation for leukemia in 73 unrelated persons. In 5 (7%) of the 68 individuals in whom the KIR2DL1 gene was present and in 10 (15%) of the 67 in whom KIR3DL1 was present, the corresponding receptor was not expressed by NK cells, as determined by flow cytometry analysis. In contrast, one or both allelic forms of KIR2DL2/KIR2DL3 were expressed by a high proportion of NK cells in all 73 individuals. However if both KIR2DL2 and KIR2DL3 genes were present, KIR2DL3 was preferentially expressed, as transcripts of KIR2DL2 was not detectable by RT-PCR in 42% of these individuals. In total, repertoire assessment for the four KIRs by genotyping vs phenotyping was not in complete agreement in 18 (25%) of the 73 individuals. Furthermore, among the samples that tested positive for the expression of a certain KIR gene, the levels of transcripts and surface expression varied considerably as measured by both real-time quantitative PCR and flow cytometry analysis. Extension of this comparative analysis to include all 12 KIR family members showed that KIR2DL3 and KIR3DL2 were the only genes whose transcripts were consistently detectable. These results caution the use of genotyping alone for donor selection or leukemia-relapse prognostication because some KIRs may be expressed at a very low level. PMID- 15879160 TI - Hepatocellular carcinoma associated with acquired von Willebrand disease and extreme thrombocytosis. PMID- 15879161 TI - The effect of cyclosporin A on airway cell proinflammatory signaling and pneumonia. AB - Cyclosporin A (CsA) blocks T cell activation by interfering with the Ca2+ dependent phosphatase, calcineurin. Proinflammatory responses to bacteria that are activated by Ca2+-fluxes in airway cells are a potential target for CsA. Although local immunosuppression may be advantageous to control airway inflammation, it could also increase susceptibility to bacterial pneumonia and invasive infection. As aerosolized CsA is currently under study in lung transplantation, we examined its direct effects on airway cells as well as in a murine model of pneumonia. Epithelial interleukin-6 production was very effectively inhibited by CsA, whereas CXCL8 production, the major PMN chemokine, was only modestly diminished. Responses to a TLR2 agonist Pam3Cys were more sensitive to CsA inhibition than those activated by Pseudomonas aeruginosa. CsA substantially blocked activation of nuclear factor of activated T cells and cAMP responsive element-binding protein (P<0.001), inhibited CCAAT/enhancer-binding protein by 50% (P<0.05), and minimally blocked activator protein-1 and nuclear factor-kappaB responses to bacteria in epithelial cells. The in vitro effects were confirmed in a mouse model of P. aeruginosa infection with similar rates of PMN recruitment, pneumonia and mortality in CsA treated and control mice. These studies indicate that airway epithelial signaling is a potential target for CsA, and such local immunosuppression may not increase susceptibility to invasive infection. PMID- 15879162 TI - Intravascular infusions of plasma into fetal sheep cause arterial and venous hypertension. AB - Fetal volume control is driven by an equilibrium between fetal and maternal hydrostatic and oncotic pressures in the placenta. Renal contributions to blood volume regulation are minor because the fetal kidneys cannot excrete fluid from the fetal compartment. We hypothesized that an increase in fetal plasma protein would lead to an increase in plasma oncotic pressure, resulting in an increase in fetal arterial and venous pressures and decreased angiotensin levels. Plasma or lactated Ringer solution was infused into each of five twin fetuses. After 7 days, fetal protein concentration was 71.2 +/- 4.2 g/l in the plasma-infused fetuses compared with 35.7 +/- 6.3 g/l in the lactated Ringer-solution-infused fetuses. Arterial pressure was 68.0 +/- 3.6 compared with 43.4 +/- 1.9 mmHg in the lactated Ringer solution-infused fetuses (P < 0.0003), whereas venous pressure was 4.8 +/- 0.3 mmHg in the plasma-infused fetuses compared with 3.3 +/- 0.4 mmHg in the lactated Ringer solution-infused fetuses (P < 0.036). Six fetuses were studied on days 0, 7, and 14 of plasma protein infusion. Fetal protein concentration increased from 31.1 +/- 1.5 to 84.8 +/- 3.8 g/l after 14 days (P < 0.01), and arterial pressure increased from 43.1 +/- 1.8 to 69.1 +/- 4.1 mmHg (P < 0.01). Venous pressure increased from 3.0 +/- 0.4 to 6.2 +/- 1.3 mmHg (P < 0.05). Fetal heart rate did not change. Angiotensin II concentration decreased, from 24.6 +/- 5.6 to 2.9 +/- 1.3 pg/l, after 14 days (P < 0.01). Fetal plasma infusions resulted in fetal arterial and venous hypertensions that could not be corrected by reductions in angiotensin II levels. PMID- 15879163 TI - Caffeine increases time to fatigue by maintaining force and not by altering firing rates during submaximal isometric contractions. AB - Caffeine increases time to fatigue [limit of endurance (T(lim))] during submaximal isometric contractions without altering whole muscle activation or neuromuscular junction transmission. We used 10 male volunteers in a randomized, double-blind, repeated-measures experiment to examine single motor unit firing rates during intermittent submaximal contractions and to determine whether administering caffeine increased T(lim) by maintaining higher firing rates. On 2 separate days, subjects performed intermittent 50% maximal voluntary contractions of the quadriceps to T(lim), 1 h after ingesting a caffeine (6 mg/kg) or placebo capsule. Average motor unit firing rates recorded with tungsten microelectrodes were constant for the duration of contractions. Caffeine increased average T(lim) by 20.5 +/- 8.1% (P < 0.05) compared with placebo conditions. This increase was due to seven subjects, termed responders, who increased T(lim) significantly. Two other subjects showed no response, and a third had a shorter T(lim). Neither the increased T(lim) nor the responders' performance could be explained by alterations in firing rates or other neuromuscular variables. However, the amplitude of the evoked twitch and its maximal instantaneous rate of relaxation did not decline to the same degree in the caffeine trial of the responders; this resulted in values 20 and 30% higher at the time point matching the end of the placebo trial (P < 0.05). The amplitude of the evoked twitch and the maximal instantaneous rate of relaxation were linearly correlated (caffeine r = 0.72, placebo r = 0.80, both P < 0.001), suggesting that the increase in T(lim) may be partially explained by caffeine's effects on calcium reuptake and twitch force. PMID- 15879164 TI - The effect of dynamic knee-extension exercise on patellar tendon and quadriceps femoris muscle glucose uptake in humans studied by positron emission tomography. AB - Both tendon and peritendinous tissue show evidence of metabolic activity, but the effect of acute exercise on substrate turnover is unknown. We therefore examined the influence of acute exercise on glucose uptake in the patellar and quadriceps tendons during dynamic exercise in humans. Glucose uptake was measured in five healthy men in the patellar and quadriceps tendons and the quadriceps femoris muscle at rest and during dynamic knee-extension exercise (25 W) using positron emission tomography and [18F]-2-fluoro-2-deoxy-D-glucose ([18F]FDG). Glucose uptake index was calculated by dividing the tissue activity with blood activity of [18F]FDG. Exercise increased glucose uptake index by 77% in the patellar tendon (from 0.30 +/- 0.09 to 0.51 +/- 0.16, P = 0.03), by 106% in the quadriceps tendon (from 0.37 +/- 0.15 to 0.75 +/- 0.36, P = 0.02), and by 15-fold in the quadriceps femoris muscle (from 0.31 +/- 0.11 to 4.5 +/- 1.7, P = 0.005). The exercise-induced increase in the glucose uptake in neither tendon correlated with the increase in glucose uptake in the quadriceps muscle (r = -0.10, P = 0.87 for the patellar tendon and r = -0.30, P = 0.62 for the quadriceps tendon). These results show that tendon glucose uptake is increased during exercise. However, the increase in tendon glucose uptake is less pronounced than in muscle and the increases are uncorrelated. Thus tendon glucose uptake is likely to be regulated by mechanisms independently of those regulating skeletal muscle glucose uptake. PMID- 15879165 TI - Muscle endurance is greater for old men compared with strength-matched young men. AB - The purpose was to compare the time to task failure for a sustained isometric contraction performed at a submaximal intensity with the elbow flexor muscles by young and old men who were matched for strength. Eight young men (18-31 yr) and eight old men (67-76 yr) sustained an isometric contraction at 20% of maximal voluntary contraction (MVC) torque until the target torque could no longer be achieved for at least 5 s. The maximal torque exerted at the wrist was similar for the young and old men before the fatiguing task (65.9 +/- 8.0 vs. 65.4 +/- 8.7 N x m; P > 0.05), and they experienced similar reductions in MVC torque after the fatiguing contraction (31.4 +/- 10.6%; P < 0.05). The time to task failure was longer for the old men (22.6 +/- 7.4 min) compared with the strength-matched young men (13.0 +/- 5.2 min; P < 0.05), despite each group sustaining a similar torque during the fatiguing contraction (P > 0.05). The increases in torque fluctuations, electromyographic (EMG) bursting activity, and heart rate were greater for young men compared with the old men, and they were less at task failure for the old men (P < 0.05). Mean arterial pressure increased at a similar rate for both groups of men (P > 0.05), whereas the averaged EMG activity and rating of perceived exertion reached similar values at task failure for the young and old men (P > 0.05). These findings indicate that the longer time to task failure for the old men when performing the submaximal contraction was not due the absolute target torque exerted during the contraction. PMID- 15879166 TI - Modification of alterations in cardiac function and sarcoplasmic reticulum by vanadate in ischemic-reperfused rat hearts. AB - To study the cardioprotective effects of vanadate on ischemia-reperfusion (I/R) injury, isolated rat hearts perfused at constant flow were subjected to global ischemia for 30 min followed by reperfusion for 30 min. In this experimental model, I/R markedly decreased ventricular developed pressure and increased end diastolic pressure. Pretreatment of hearts with 4 microM vanadate attenuated I/R induced cardiac dysfunction. The reduction in sarcoplasmic reticulum (SR) Ca2+ uptake and Ca2+ release, as well as SR protein contents for Ca2+-pump ATPase and Ca2+-release channel, was also prevented by vanadate. Pretreatment of hearts with an antioxidant mixture containing superoxide dismutase + catalase exerted effects similar to those of vanadate in I/R hearts. Postischemic treatment of hearts with vanadate or superoxide dismutase + catalase also had beneficial effects on I/R induced changes in cardiac performance and SR function. Alterations in cardiac function and SR Ca2+ transport due to an oxyradical-generating system (xanthine + xanthine oxidase) or an oxidant (H2O2) were attenuated by treatment with vanadate. These results suggest that vanadate may exert beneficial effects on cardiac performance and SR function in I/R hearts because of its antioxidant action. PMID- 15879167 TI - Influence of gender, menstrual phase, and oral contraceptive use on immunological changes in response to prolonged cycling. AB - This study determined the influence of gender, menstrual phase (MP), and oral contraceptive (OC) use on immunological changes in response to endurance exercise. Twelve women and 11 men similar in age, aerobic power, and activity level cycled for 90 min at 65% maximal aerobic power. Women were OC users (n = 6) or nonusers (NOC) and cycled during the follicular (Fol) and the luteal (Lut) phases. Venous blood was collected before and after exercise to determine leukocyte counts, IL-6 concentrations, and cortisol. Higher resting levels of neutrophils (approximately 1.5-fold) and cortisol (approximately 2.5-fold) were found in OC vs. NOC and men. Exercise-induced immune cell count and IL-6 changes were similar between men and NOC, except for an approximately 38% greater lymphocyte response in NOC vs. men (P = 0.07). Neutrophil, monocyte, and lymphocyte responses to exercise during Lut in OC were greater than during Fol and also greater than the responses in men (P < or = 0.003). Changes in immune cell counts were consistently greater during Lut in OC vs. NOC, regardless of MP, but only neutrophil responses reached statistical significance (P = 0.01). The exercise-induced change in IL-6 was approximately 80% greater in NOC vs. OC during Fol (P = 0.06), but it was similar between these groups during Lut. Cortisol changes with exercise were not different between groups or MP. These results highlight the necessity to control for gender, and in particular OC use, when designing studies evaluating exercise and immunology. PMID- 15879168 TI - Influence of muscle glycogen availability on ERK1/2 and Akt signaling after resistance exercise in human skeletal muscle. AB - Two pathways that have been implicated for cellular growth and development in response to muscle contraction are the extracellular signal-regulated kinase (ERK1/2) and Akt signaling pathways. Although these pathways are readily stimulated after exercise, little is known about how nutritional status may affect stimulation of these pathways in response to resistance exercise in human skeletal muscle. To investigate this, experienced cyclists performed 30 repetitions of knee extension exercise at 70% of one repetition maximum after a low (2%) or high (77%) carbohydrate (LCHO or HCHO) diet, which resulted in low or high (approximately 174 or approximately 591 mmol/kg dry wt) preexercise muscle glycogen content. Muscle biopsies were taken from the vastus lateralis before, approximately 20 s after, and 10 min after exercise. ERK1/2 and p90 ribosomal S6 kinase phosphorylation increased (P < or = 0.05) 10 min after exercise, regardless of muscle glycogen availability. Akt phosphorylation was elevated (P < 0.05) 10 min after exercise in the HCHO trial but was unaffected after exercise in the LCHO trial. Mammalian target of rapamycin phosphorylation was similar to that of Akt during each trial; however, change or lack of change was not significant. In conclusion, the ERK1/2 pathway appears to be unaffected by muscle glycogen content. However, muscle glycogen availability appears to contribute to regulation of the Akt pathway, which may influence cellular growth and adaptation in response to resistance exercise in a low-glycogen state. PMID- 15879169 TI - Heat extraction through the palm of one hand improves aerobic exercise endurance in a hot environment. AB - In situations where the accumulation of internal heat limits physical performance, enhanced heat extraction from the body should improve performance capacity. The combined application of local subatmospheric pressure (35-45 mmHg) to an entire hand (to increase blood volume) and a heat sink (18-22 degrees C) to the palmar surface were used to draw heat out of the circulating blood. Subjects walked uphill (5.63 km/h) on a treadmill in a 40 degree C environment. Slopes of the treadmill were held constant during paired experimental trials (with and without the device). Heat extraction attenuated the rate of esophageal temperature rise during exercise (2.1 +/- 0.4 degrees and 2.9 +/- 0.5 degrees C/h, mean +/- SE, with and without the device, respectively; n = 8) and increased exercise duration (46.1 +/- 3.4 and 32.3 +/- 1.7 min with and without the device, respectively; n = 18). Hand cooling alone had little effect on exercise duration (34.1 +/- 3.0, 38.0 +/- 3.5, and 57.0 +/- 6.4 min, for control, cooling only, and cooling, and subatmospheric pressure, respectively; n = 6). In a longer term study, nine subjects participated in two or four trials per week for 8 wk. The individual workloads (treadmill slope) were varied weekly. Use of the device had a beneficial effect on exercise endurance at all workloads, but the benefit proportionally decreased at higher workloads. It is concluded that heat can be efficiently removed from the body by using the described technology and that such treatment can provide a substantial performance benefit in thermally stressful conditions. PMID- 15879170 TI - Acute intermittent hypoxia improves rat myocardium tolerance to ischemia. AB - In this study, we investigated the influence of depth and duration of intermittent hypoxia (IH) on the infarct size development in isolated rat heart. The role of nitric oxide synthase (NOS) and ATP-sensitive K+ (K(ATP)) channel was also studied. Wistar male rats were exposed to IH [repetitive cycles of 1 min, 40 s with inspired oxygen fraction (FI(O2)), 5 or 10%, followed by 20-s normoxia], during 30 min or 4 h. Another group was exposed to 4 h of continuous hypoxia with 10% FI(O2). Twenty-four hours later, their hearts were isolated and subjected to a 30-min no-flow global ischemia-120-min reperfusion sequence. For some hearts, N(omega)-nitro-L-arginine methyl ester (L-NAME) (a nonselective inhibitor of NOS) or 5-hydroxydecanoic acid (5-HD) (a selective mitochondrial K(ATP) blocker) was infused before ischemia. Infarct size (in percentage of ventricles) was significantly reduced by prior IH for 4 h (10% FI(O2)) (21.8 +/- 3.1 vs. 33.5 +/- 2.5% in sham group). This effect was abolished by L-NAME or 5-HD. Infarct size was not different in groups subjected to either 30 min of IH or to continuous hypoxia compared with sham group. In contrast, IH for 4 h (5% FI(O2)) significantly increased infarct size (45.1 +/- 3.6 vs. 33.5 +/- 2.5% in sham group). Acute IH for 4 h with a minimal FI(O2) of 10% induced a delayed preconditioning against myocardial infarction in the rat, which was abolished by NOS inhibition and mitochondrial K(ATP) channel blockade. Depth, duration, and intermittence of hypoxia appeared to be critical for cardioprotection to occur. PMID- 15879171 TI - Carbohydrate supplementation improves time-trial cycle performance during energy deficit at 4,300-m altitude. AB - Carbohydrate supplementation (CHOS) typically improves prolonged time-trial (TT) performance at sea level (SL). This study determined whether CHOS also improves TT performance at high altitude (ALT; 4,300 M) despite increased hypoxemia and while in negative energy balance (approximately 1,250 kcal/day). Two groups of fasting, fitness-matched men performed a 720-kJ cycle TT at SL and while living at ALT on days 3 (ALT3) and 10 (ALT10). Eight men drank a 10% carbohydrate solution (0.175 g/kg body wt) and eight drank a placebo (PLA; double blind) at the start of and every 15 min of the TT. Blood glucose during each TT was higher (P < 0.05) for CHOS than for PLA. At SL, TT duration (approximately 59 min) and watts (approximately 218 or approximately 61% of peak watts; %SL Wpeak) were similar for both groups. At ALT, the TT was longer for both groups (P < 0.01) but was shorter for CHOS than for PLA on ALT3 (means +/- SE: 80 +/- 7 vs. 105 +/- 9 min; P < 0.01) and ALT10 (77 +/- 7 vs. 90 +/- 5 min; P < 0.01). At ALT, %SL Wpeak was reduced (P < 0.01) with the reduction on ALT3 being larger for PLA (to 33 +/- 3%) than for CHOS (to 43 +/- 2%; P < 0.05). On ALT3, O2 saturation fell similarly from 84 +/- 2% at rest to 73 +/- 1% during the TT for both groups (P < 0.05), and on ALT10 O2 saturation fell more (P < 0.02) for CHOS (91 +/- 1 to 76 +/- 2%) than for PLA (90 +/- 1 to 81 +/- 1%). %SL Wpeak and O2 saturation were inversely related during the TT for both groups at ALT (r > or = -0.76; P < or = 0.03). It was concluded that, despite hypoxemia exacerbated by exercise, CHOS greatly improved TT performance at ALT in which there was a negative energy balance. PMID- 15879172 TI - The rotational spectrum and structure of the HOOO radical. AB - The adduct of the hydroxyl radical with oxygen has been studied theoretically, in connection with atmospheric reactions, but its stability and structure remained an open question. Pure rotational spectra of the HOOO and DOOO radicals have now been observed in a supersonic jet by using a Fourier-transform microwave spectrometer with a pulsed discharge nozzle. The molecular constants extracted from 12 rotational transitions with fine and hyperfine splittings support a trans planar molecular structure, in contrast to the cis planar structure predicted by most ab initio calculations. The bond linking the HO and O2 moieties is fairly long (1.688 angstroms) and comparable to the F-O bond in the isoelectronic FOO radical. PMID- 15879173 TI - Community proteomics of a natural microbial biofilm. AB - Using genomic and mass spectrometry-based proteomic methods, we evaluated gene expression, identified key activities, and examined partitioning of metabolic functions in a natural acid mine drainage (AMD) microbial biofilm community. We detected 2033 proteins from the five most abundant species in the biofilm, including 48% of the predicted proteins from the dominant biofilm organism, Leptospirillum group II. Proteins involved in protein refolding and response to oxidative stress appeared to be highly expressed, which suggests that damage to biomolecules is a key challenge for survival. We validated and estimated the relative abundance and cellular localization of 357 unique and 215 conserved novel proteins and determined that one abundant novel protein is a cytochrome central to iron oxidation and AMD formation. PMID- 15879174 TI - Extension of murine life span by overexpression of catalase targeted to mitochondria. AB - To determine the role of reactive oxygen species in mammalian longevity, we generated transgenic mice that overexpress human catalase localized to the peroxisome, the nucleus, or mitochondria (MCAT). Median and maximum life spans were maximally increased (averages of 5 months and 5.5 months, respectively) in MCAT animals. Cardiac pathology and cataract development were delayed, oxidative damage was reduced, H2O2 production and H2O2-induced aconitase inactivation were attenuated, and the development of mitochondrial deletions was reduced. These results support the free radical theory of aging and reinforce the importance of mitochondria as a source of these radicals. PMID- 15879175 TI - A mutation in the TRPC6 cation channel causes familial focal segmental glomerulosclerosis. AB - Focal and segmental glomerulosclerosis (FSGS) is a kidney disorder of unknown etiology, and up to 20% of patients on dialysis have been diagnosed with it. Here we show that a large family with hereditary FSGS carries a missense mutation in the TRPC6 gene on chromosome 11q, encoding the ion-channel protein transient receptor potential cation channel 6 (TRPC6). The proline-to-glutamine substitution at position 112, which occurs in a highly conserved region of the protein, enhances TRPC6-mediated calcium signals in response to agonists such as angiotensin II and appears to alter the intracellular distribution of TRPC6 protein. Previous work has emphasized the importance of cytoskeletal and structural proteins in proteinuric kidney diseases. Our findings suggest an alternative mechanism for the pathogenesis of glomerular disease. PMID- 15879176 TI - An endless frontier postponed. PMID- 15879177 TI - Public health. A heavyweight battle over CDC's obesity forecasts. PMID- 15879178 TI - Astronomy. Picture-perfect planet on course for the history books. PMID- 15879179 TI - Computing. IBM offers free number crunching for humanitarian research projects. PMID- 15879181 TI - European policy. Panel gives thumbs-down to European Institute of Technology. PMID- 15879180 TI - Science resources. Chemists want NIH to curtail database. PMID- 15879183 TI - NASA. U.S. lawmakers call for new earth science strategy. PMID- 15879182 TI - Genomics. Celera to end subscriptions and give data to public GenBank. PMID- 15879184 TI - Department of Energy. Two-thirds of Senate backs more research. PMID- 15879185 TI - Neuroscience. The dark side of glia. PMID- 15879186 TI - Embryology. Embryologists polarized over early cell fate determination. PMID- 15879187 TI - Technology. Electronic paper: a revolution about to unfold? PMID- 15879188 TI - Technology. Shrinking dimensions spur research into ever-slimmer batteries. PMID- 15879189 TI - Atmospheric science. Changes in the sun may sway the tropical monsoon. PMID- 15879190 TI - Calling on scientists to fight budget cuts. PMID- 15879192 TI - Memo to NASA: finish what you start. PMID- 15879191 TI - Establishing indicators for biodiversity. PMID- 15879193 TI - The end of a Chilean institute. PMID- 15879194 TI - Comment on "Energetics of hydrogen bond network rearrangements in liquid water". PMID- 15879196 TI - Essays on science and society. Pleistocene Park: return of the mammoth's ecosystem. PMID- 15879197 TI - Neuroscience. Attractors in memory. PMID- 15879198 TI - Neuroscience. Matching at the synapse. PMID- 15879199 TI - Cell biology. Wnt signaling glows with RNAi. PMID- 15879200 TI - Retrospective: Stanley Joel Korsmeyer (1950-2005). PMID- 15879201 TI - Atmosphere. Air pollution-related illness: effects of particles. PMID- 15879202 TI - Atmospheric science. In search of balance. PMID- 15879203 TI - Geochemistry. The paradox of mantle redox. PMID- 15879205 TI - Distributed computing. Grassroots supercomputing. PMID- 15879206 TI - Distributed computing. Grid sport: competitive crunching. PMID- 15879207 TI - Distributed computing. Data-bots chart the Internet. PMID- 15879208 TI - Service-oriented science. AB - New information architectures enable new approaches to publishing and accessing valuable data and programs. So-called service-oriented architectures define standard interfaces and protocols that allow developers to encapsulate information tools as services that clients can access without knowledge of, or control over, their internal workings. Thus, tools formerly accessible only to the specialist can be made available to all; previously manual data-processing and analysis tasks can be automated by having services access services. Such service-oriented approaches to science are already being applied successfully, in some cases at substantial scales, but much more effort is required before these approaches are applied routinely across many disciplines. Grid technologies can accelerate the development and adoption of service-oriented science by enabling a separation of concerns between discipline-specific content and domain-independent software and hardware infrastructure. PMID- 15879209 TI - Cyberinfrastructure for e-Science. AB - Here we describe the requirements of an e-Infrastructure to enable faster, better, and different scientific research capabilities. We use two application exemplars taken from the United Kingdom's e-Science Programme to illustrate these requirements and make the case for a service-oriented infrastructure. We provide a brief overview of the UK "plug-and-play composable services" vision and the role of semantics in such an e-Infrastructure. PMID- 15879210 TI - Cyberinfrastructure: empowering a "third way" in biomedical research. AB - Biomedicine has experienced explosive growth, fueled in parts by the substantial increase of government support, continued development of the biotechnology industry, and the increasing adoption of molecular-based medicine. At its core, it is composed of fiercely independent, innovative, entrepreneurial individuals, organizations, and institutions. The field has developed unprecedented capacity to characterize biologic systems at their most fundamental levels with the use of tools and technologies almost unimaginable a generation ago. Biomedicine is at the precipice of unlocking the very essence of biologic life and enabling a new generation of medicine. Development and deployment of cyberinfrastructure may prove to be on the critical path to obtaining these goals. PMID- 15879211 TI - Changes in Earth's albedo measured by satellite. AB - NASA global satellite data provide observations of Earth's albedo, i.e., the fraction of incident solar radiation that is reflected back to space. The satellite data show that the last four years are within natural variability and fail to confirm the 6% relative increase in albedo inferred from observations of earthshine from the moon. Longer global satellite records will be required to discern climate trends in Earth's albedo. PMID- 15879212 TI - The optical resonances in carbon nanotubes arise from excitons. AB - Optical transitions in carbon nanotubes are of central importance for nanotube characterization. They also provide insight into the nature of excited states in these one-dimensional systems. Recent work suggests that light absorption produces strongly correlated electron-hole states in the form of excitons. However, it has been difficult to rule out a simpler model in which resonances arise from the van Hove singularities associated with the one-dimensional band [corrected] structure of the nanotubes. Here, two-photon excitation spectroscopy bolsters the exciton picture. We found binding energies of approximately 400 millielectron volts for semiconducting single-walled nanotubes with 0.8-nanometer diameters. The results demonstrate the dominant role of many-body interactions in the excited-state properties of one-dimensional systems. PMID- 15879213 TI - Zircon thermometer reveals minimum melting conditions on earliest Earth. AB - Ancient zircons from Western Australia's Jack Hills preserve a record of conditions that prevailed on Earth not long after its formation. Widely considered to have been a uniquely violent period geodynamically, the Hadean Eon [4.5 to 4.0 billion years ago (Ga)] has recently been interpreted by some as far more benign-possibly even characterized by oceans like those of the present day. Knowledge of the crystallization temperatures of the Hadean zircons is key to this debate. A thermometer based on titanium content revealed that these zircons cluster strongly at approximately 700 degrees C, which is indistinguishable from temperatures of granitoid zircon growth today and strongly suggests a regulated mechanism producing zircon-bearing rocks during the Hadean. The temperatures substantiate the existence of wet, minimum-melting conditions within 200 million years of solar system formation. They further suggest that Earth had settled into a pattern of crust formation, erosion, and sediment recycling as early as 4.35 Ga. PMID- 15879214 TI - From dimming to brightening: decadal changes in solar radiation at Earth's surface. AB - Variations in solar radiation incident at Earth's surface profoundly affect the human and terrestrial environment. A decline in solar radiation at land surfaces has become apparent in many observational records up to 1990, a phenomenon known as global dimming. Newly available surface observations from 1990 to the present, primarily from the Northern Hemisphere, show that the dimming did not persist into the 1990s. Instead, a widespread brightening has been observed since the late 1980s. This reversal is reconcilable with changes in cloudiness and atmospheric transmission and may substantially affect surface climate, the hydrological cycle, glaciers, and ecosystems. PMID- 15879215 TI - Do satellites detect trends in surface solar radiation? AB - Long-term variations in solar radiation at Earth's surface (S) can affect our climate, the hydrological cycle, plant photosynthesis, and solar power. Sustained decreases in S have been widely reported from about the year 1960 to 1990. Here we present an estimate of global temporal variations in S by using the longest available satellite record. We observed an overall increase in S from 1983 to 2001 at a rate of 0.16 watts per square meter (0.10%) per year; this change is a combination of a decrease until about 1990, followed by a sustained increase. The global-scale findings are consistent with recent independent satellite observations but differ in sign and magnitude from previously reported ground observations. Unlike ground stations, satellites can uniformly sample the entire globe. PMID- 15879216 TI - The Holocene Asian monsoon: links to solar changes and North Atlantic climate. AB - A 5-year-resolution absolute-dated oxygen isotope record from Dongge Cave, southern China, provides a continuous history of the Asian monsoon over the past 9000 years. Although the record broadly follows summer insolation, it is punctuated by eight weak monsoon events lasting approximately 1 to 5 centuries. One correlates with the "8200-year" event, another with the collapse of the Chinese Neolithic culture, and most with North Atlantic ice-rafting events. Cross correlation of the decadal- to centennial-scale monsoon record with the atmospheric carbon-14 record shows that some, but not all, of the monsoon variability at these frequencies results from changes in solar output. PMID- 15879217 TI - Computational thermostabilization of an enzyme. AB - Thermostabilizing an enzyme while maintaining its activity for industrial or biomedical applications can be difficult with traditional selection methods. We describe a rapid computational approach that identified three mutations within a model enzyme that produced a 10 degrees C increase in apparent melting temperature T(m) and a 30-fold increase in half-life at 50 degrees C, with no reduction in catalytic efficiency. The effects of the mutations were synergistic, giving an increase in excess of the sum of their individual effects. The redesigned enzyme induced an increased, temperature-dependent bacterial growth rate under conditions that required its activity, thereby coupling molecular and metabolic engineering. PMID- 15879218 TI - Swimming against the flow: a mechanism of zooplankton aggregation. AB - Zooplankton reside in a constantly flowing environment. However, information about their response to ambient flow has remained elusive, because of the difficulties of following the individual motions of these minute, nearly transparent animals in the ocean. Using a three-dimensional acoustic imaging system, we tracked >375,000 zooplankters at two coastal sites in the Red Sea. Resolution of their motion from that of the water showed that the animals effectively maintained their depth by swimming against upwelling and downwelling currents moving at rates of up to tens of body lengths per second, causing their accumulation at frontal zones. This mechanism explains how oceanic fronts become major feeding grounds for predators and targets for fishermen. PMID- 15879219 TI - A synaptonemal complex protein promotes homology-independent centromere coupling. AB - We describe a process in meiotic cells of budding yeast in which chromosomes become joined together in pairs at their centromeres independent of chromosomal homology. These centromeric interactions depend on the synaptonemal complex component Zip1. During meiosis in wild-type diploids, centromere couples are initially nonhomologous and then undergo switching until all couples involve homologs. This transition to homologous coupling depends on Spo11, a protein required for the initiation of meiotic recombination. Regions of synaptonemal complex assembled early in meiosis are often centromere-associated. We propose that centromere coupling facilitates homolog pairing and promotes synapsis initiation. PMID- 15879220 TI - Attractor dynamics in the hippocampal representation of the local environment. AB - Memories are thought to be attractor states of neuronal representations, with the hippocampus a likely substrate for context-dependent episodic memories. However, such states have not been directly observed. For example, the hippocampal place cell representation of location was previously found to respond continuously to changes in environmental shape alone. We report that exposure to novel square and circular environments made of different materials creates attractor representations for both shapes: Place cells abruptly and simultaneously switch between representations as environmental shape changes incrementally. This enables study of attractor dynamics in a cognitive representation and may correspond to the formation of distinct contexts in context-dependent memory. PMID- 15879221 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. AB - The purpose of this announcement is to effect the valid publication of the following new names and new combinations under the procedure described in the Bacteriological Code (1990 Revision). Authors and other individuals wishing to have new names and/or combinations included in future lists should send three copies of the pertinent reprint or photocopies thereof to the IJSEM Editorial Office for confirmation that all of the other requirements for valid publication have been met. It is also a requirement of IJSEM and the ICSP that authors of new species, new subspecies and new combinations provide evidence that types are deposited in two recognized culture collections in two different countries (i.e. documents certifying deposition and availability of type strains). It should be noted that the date of valid publication of these new names and combinations is the date of publication of this list, not the date of the original publication of the names and combinations. The authors of the new names and combinations are as given below, and these authors' names will be included in the author index of the present issue and in the volume author index. Inclusion of a name on these lists validates the publication of the name and thereby makes it available in bacteriological nomenclature. The inclusion of a name on this list is not to be construed as taxonomic acceptance of the taxon to which the name is applied. Indeed, some of these names may, in time, be shown to be synonyms, or the organisms may be transferred to another genus, thus necessitating the creation of a new combination. PMID- 15879222 TI - Notification that new names and new combinations have appeared in volume 55, part 1, of the IJSEM. AB - This listing of names published in a previous issue of the IJSEM is provided as a service to bacteriology to assist in the recognition of new names and new combinations. This procedure was proposed by the Judicial Commission [Minute 11(ii), Int J Syst Bacteriol 41 (1991), p. 185]. The names given herein are listed according to the Rules of priority (i.e. page number and order of valid publication of names in the original articles). Taxonomic opinions included in this List (i.e. the creation of synonyms or the emendation of circumscriptions) cannot be considered as validly published nor, in any other way, approved by the International Committee on Systematics of Prokaryotes and its Judicial Commission. PMID- 15879223 TI - Bundling the forces in systematists. PMID- 15879224 TI - Desulfurococcus fermentans sp. nov., a novel hyperthermophilic archaeon from a Kamchatka hot spring, and emended description of the genus Desulfurococcus. AB - An obligately anaerobic, hyperthermophilic, organoheterotrophic archaeon, strain Z-1312(T), was isolated from a freshwater hot spring of the Uzon caldera (Kamchatka Peninsula, Russia). The cells were regular cocci, 1-4 microm in diameter, with one long flagellum. The cell envelope was composed of a globular layer attached to the cytoplasmic membrane. The temperature range for growth was 63-89 degrees C, with an optimum between 80 and 82 degrees C. The pH range for growth at 80 degrees C was 4.8-6.8, with an optimum at pH 6.0. Strain Z-1312(T) grew by hydrolysis and/or fermentation of a wide range of polymeric and monomeric substrates, including agarose, amygdalin, arabinose, arbutin, casein hydrolysate, cellulose (filter paper, microcrystalline cellulose, carboxymethyl cellulose), dextran, dulcitol, fructose, lactose, laminarin, lichenan, maltose, pectin, peptone, ribose, starch and sucrose. No growth was detected on glucose, xylose, mannitol or sorbitol. Growth products when sucrose or starch were used as the substrate were acetate, H(2) and CO(2). Elemental sulfur, thiosulfate and nitrate added as potential electron acceptors for anaerobic respiration did not stimulate growth when tested with starch as the substrate. H(2) at 100 % in the gas phase did not inhibit growth on starch or peptone. The G+C content of the DNA was 42.5 mol%. 16S rRNA gene sequence analysis placed the isolated strain Z-1312(T) as a member of the genus Desulfurococcus, where it represented a novel species, for which the name Desulfurococcus fermentans sp. nov. (type strain Z-1312(T) = DSM 16532 (T) = VKM V-2316(T)) is proposed. PMID- 15879225 TI - Desulfonatronum cooperativum sp. nov., a novel hydrogenotrophic, alkaliphilic, sulfate-reducing bacterium, from a syntrophic culture growing on acetate. AB - A novel alkaliphilic, sulfate-reducing bacterium was isolated from a syntrophic acetate-decomposing community enriched from samples of the soda lake Khadin, Tuva, Russia; the isolate was designated strain Z-7999(T). Cells of strain Z 7999(T) were vibrioid, Gram-negative, 0.4-0.5 x 1.0-2.5 microm and motile by means of a polar flagellum. The temperature range for growth was 15-40 degrees C, with an optimum of 35-38 degrees C. The pH range for growth was 6.7-10.3, with an optimum of pH 8.0-9.0. The NaCl concentration range for growth was 1-80 g l(-1). The novel isolate was obligately anaerobic, was alkaliphilic with a broad pH range and had an obligate requirement for carbonate ions in the growth medium. In the presence of sulfate as electron acceptor, it grew with hydrogen, formate and lactate. It was not able to ferment sugars, organic acids, amino acids or peptides. During growth on formate, strain Z-7999(T) reduced sulfite and thiosulfate to sulfide. It was able to grow lithoheterotrophically with sulfate and formate when acetate was added as a carbon source for biosynthesis of biomass. The G + C content of the genomic DNA of strain Z-7999(T) was 56.5 mol%. Results of comparative 16S rRNA gene sequence analyses revealed that strain Z 7999(T) was part of the delta-Proteobacteria and clustered with other members of the genus Desulfonatronum (similarity values of 95.2 and 95.3 % to Desulfonatronum lacustre and Desulfonatronum thiodismutans, respectively). DNA DNA hybridization with D. lacustre was 37 %. On the basis of physiological and phylogenetic data, it is proposed that strain Z-7999(T) (= DSM 16749(T) = VKM B 2329(T)) should be placed in the genus Desulfonatronum as a representative of a novel species, Desulfonatronum cooperativum sp. nov. PMID- 15879226 TI - Psychrobacter aquimaris sp. nov. and Psychrobacter namhaensis sp. nov., isolated from sea water of the South Sea in Korea. AB - Two Gram-negative, non-motile, non-spore-forming, slightly halophilic bacterial strains, SW-210(T) and SW-242(T), were isolated from sea water of the South Sea in Korea, and were characterized taxonomically by means of a polyphasic approach. The two isolates grew optimally at 25-30 degrees C and in the presence of 2-3 % (w/v) NaCl. Strains SW-210(T) and SW-242(T) contained Q-8 as the predominant respiratory lipoquinone and C(18 : 1)omega9c as the major fatty acid. The DNA G+C contents of strains SW-210(T) and SW-242(T) were 43.2 and 45.3 mol%, respectively. The 16S rRNA gene sequences of strains SW-210(T) and SW-242(T) were 98.9 % similar, and the mean DNA-DNA relatedness value between them was 24 %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains SW 210(T) and SW-242(T) form distinct phylogenetic lineages within the radiation of the cluster comprising Psychrobacter species, having 16S rRNA gene sequence similarity levels of 95.9-99.2 % with respect to the type strains of Psychrobacter species. The levels of DNA-DNA relatedness between the two isolates and the type strains of 15 phylogenetically related Psychrobacter species were well below 70 %. On the basis of phenotypic and phylogenetic data and genomic distinctiveness, strains SW-210(T) and SW-242(T) were classified in the genus Psychrobacter as representing two distinct novel species, for which the names Psychrobacter aquimaris sp. nov. (type strain, SW-210(T) = KCTC 12254(T) = DSM 16329(T)) and Psychrobacter namhaensis sp. nov. (type strain, SW-242(T) = KCTC 12255(T) = DSM 16330(T)) are proposed, respectively. PMID- 15879227 TI - Muricauda flavescens sp. nov. and Muricauda aquimarina sp. nov., isolated from a salt lake near Hwajinpo Beach of the East Sea in Korea, and emended description of the genus Muricauda. AB - Four Gram-negative, non-spore-forming, slightly halophilic rods (strains SW 62(T), SW-74, SW-63(T) and SW-72) with appendages were isolated from a salt lake near Hwajinpo Beach on the East Sea in Korea, and subjected to a polyphasic taxonomic analysis. Phylogenetic analyses based on 16S rRNA gene sequences showed that strains SW-62(T), SW-74, SW-63(T) and SW-72 formed a coherent cluster with Muricauda ruestringensis. Strains SW-62(T) and SW-74 had the same 16S rRNA gene sequence, as did strains SW-63(T) and SW-72. The level of 16S rRNA gene sequence similarity between strains SW-62(T) and SW-63(T) was 97.0 %. Strains SW-62(T) and SW-63(T) exhibited 16S rRNA gene similarity levels of 96.5 and 98.3 %, respectively, with respect to M. ruestringensis DSM 13258(T). The predominant isoprenoid quinone found in the four isolates and M. ruestringensis DSM 13258(T) was MK-6. The four strains contained iso-C(17 : 0) 3-OH, iso-C(15 : 1) and iso C(15 : 0) as the major fatty acids. Their DNA G + C contents were 44.1-45.4 mol%. The levels of DNA-DNA relatedness indicated that strains SW-62(T) and SW-74 and strains SW-63(T) and SW-72 were members of two species that were different from M. ruestringensis. On the basis of phenotypic and phylogenetic data and genomic distinctiveness, strains SW-62(T) and SW-74 and strains SW-63(T) and SW-72 were placed in the genus Muricauda as two distinct novel species, for which the names Muricauda flavescens sp. nov. (type strain, SW-62(T) = KCCM 41645(T) = JCM 11812(T)) and Muricauda aquimarina sp. nov. (type strain, SW-63(T) = KCCM 41646(T) = JCM 11811(T)), respectively, are proposed. PMID- 15879228 TI - Genomic analysis of Hyphomonas neptunium contradicts 16S rRNA gene-based phylogenetic analysis: implications for the taxonomy of the orders 'Rhodobacterales' and Caulobacterales. AB - Hyphomonas neptunium is a marine prosthecate alpha-proteobacterium currently classified as a member of the order 'Rhodobacterales'. Although this classification is supported by 16S rRNA gene sequence phylogeny, 23S rRNA gene sequence analysis, concatenated ribosomal proteins, HSP70 and EF-Tu phylogenies all support classifying Hyphomonas neptunium as a member of the Caulobacterales instead. The possible reasons why the 16S rRNA gene sequence gives conflicting results in this case are also discussed. PMID- 15879229 TI - Pontibacillus marinus sp. nov., a moderately halophilic bacterium from a solar saltern, and emended description of the genus Pontibacillus. AB - A moderately halophilic, Gram-positive, rod-shaped bacterium (BH030004(T)) was isolated from a solar saltern in Korea. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain BH030004(T) belonged to the genus Pontibacillus. Chemotaxonomic data (DNA G + C content, 42 mol%; major isoprenoid quinone, MK-7; cell-wall type, A1gamma-type meso-diaminopimelic acid; major fatty acids, iso-C(15 : 0) and anteiso-C(15 : 0)) also supported the affiliation of the isolate to the genus Pontibacillus. Although the 16S rRNA gene sequence similarity between strain BH030004(T) and Pontibacillus chungwhensis DSM 16287(T) was relatively high (99.1 %), physiological properties and DNA-DNA hybridization (about 7 % DNA-DNA relatedness) allowed genotypic and phenotypic differentiation of strain BH030004(T) from the type strain of P. chungwhensis. Therefore, strain BH030004(T) represents a novel species of the genus Pontibacillus, for which the name Pontibacillus marinus sp. nov. is proposed. The type strain is BH030004(T) (=KCTC 3917(T) = DSM 16465(T)). PMID- 15879230 TI - Reclassification of [Cytophaga] marinoflava Reichenbach 1989 as Leeuwenhoekiella marinoflava gen. nov., comb. nov. and description of Leeuwenhoekiella aequorea sp. nov. AB - Five heterotrophic, aerobic, halotolerant and pigmented bacterial strains with gliding motility were isolated from Antarctic sea water; one other isolate was collected from the sea urchin Strongylocentrotus intermedius in the Gulf of Peter the Great in the Sea of Japan. 16S rRNA gene sequence analysis indicated that the strains are members of the family Flavobacteriaceae, the nearest neighbour (with 97.1 % sequence similarity) being the misclassified species [Cytophaga] marinoflava. DNA-DNA hybridization experiments and chemotaxonomic and phenotypic analyses demonstrated that the six novel isolates represent a single species distinct from [C.] marinoflava. On the basis of its separate phylogenetic lineage (the nearest neighbours show 92 % sequence similarity), [C.] marinoflava is reclassified as Leeuwenhoekiella marinoflava gen. nov., comb. nov. A second species of this new genus, Leeuwenhoekiella aequorea sp. nov., is proposed for the six novel isolates, with strain LMG 22550(T) (=CCUG 50091(T)) as the type strain. PMID- 15879231 TI - Brevibacillus levickii sp. nov. and Aneurinibacillus terranovensis sp. nov., two novel thermoacidophiles isolated from geothermal soils of northern Victoria Land, Antarctica. AB - Thirteen strains of endospore-forming bacteria were isolated from geothermal soils at Cryptogam Ridge, the north-west slope of Mt Melbourne, and at the vents and summit of Mt Rittmann in northern Victoria Land, Antarctica. 16S rRNA gene sequencing, SDS-PAGE and routine phenotypic characterization tests indicated that the seven isolates from the north-west slope of Mt Melbourne represent a novel species of Brevibacillus and that the six isolates from Cryptogam Ridge and the vents and summit of Mt Rittmann represent a novel species of Aneurinibacillus. Brevibacillus strains were not isolated from the sites at Mt Rittmann or Cryptogam Ridge and Aneurinibacillus strains were not isolated from the north west slope of Mt Melbourne. Preliminary metabolic studies revealed that L glutamic acid, although not essential for growth, was utilized by both species. The Brevibacillus species possessed an uptake system specific for L-glutamic acid, whereas the Aneurinibacillus species possessed a more general uptake system capable of transporting other related amino acids. Both species utilized a K(+) antiport system and similar energy systems for the uptake of l-glutamic acid. The rate of uptake by the Brevibacillus species type strain was 20-fold greater than that shown by the Aneurinibacillus species type strain. The names Brevibacillus levickii sp. nov. and Aneurinibacillus terranovensis sp. nov. are proposed for the novel taxa; the type strains are Logan B-1657(T) (= LMG 22481(T) = CIP 108307(T)) and Logan B-1599(T) (LMG 22483(T) = CIP 108308(T)), respectively. PMID- 15879232 TI - Owenweeksia hongkongensis gen. nov., sp. nov., a novel marine bacterium of the phylum 'Bacteroidetes'. AB - An aerobic, Gram-negative, non-fermentative, rod-shaped, motile, orange-pigmented bacterium, UST20020801(T), was isolated from sea-water samples collected from Port Shelter, Hong Kong, S.A.R., China, in August 2002. The full 16S rRNA gene sequence of this strain shared only 87.5 % similarity with its nearest relative, Crocinitomix catalasitica, a species of the family Cryomorphaceae. However, strain UST20020801(T) possessed menaquinone-6, a major respiratory quinone of members of the family Flavobacteriaceae. This strain contains unique fatty acids such as i15 : 1G, i17 : 1omega9c, 2-OH 15 : 0, 15 : 1omega6c and three unknown fatty acids of equivalent chain-length of 11.543, 13.565 and 16.582. Further analysis of its ecophysiology and biochemistry suggests that this strain represents a new genus in the phylum 'Bacteroidetes'. The name Owenweeksia hongkongensis gen. nov., sp. nov. is proposed. The type strain is UST20020801(T) (=NRRL B-23963(T) = JCM 12287(T)). PMID- 15879233 TI - Thiovirga sulfuroxydans gen. nov., sp. nov., a chemolithoautotrophic sulfur oxidizing bacterium isolated from a microaerobic waste-water biofilm. AB - A novel mesophilic, chemolithoautotrophic, sulfur-oxidizing bacterium, designated strain SO07(T), was isolated from a microaerobic waste-water biofilm. Chemolithoautotrophic growth was observed with elemental sulfur, sulfide and thiosulfate as sole electron donors and oxygen as electron acceptor. Anaerobic and heterotrophic growth were not observed. Nitrate was not used as a terminal electron acceptor. The optimum pH and temperature for growth were pH 7.5 and 30 degrees C, respectively. The major isoprenoid quinone was Q-8. The DNA G + C content of strain SO07(T) was 47.1 mol%. Phylogenetic analysis of 16S rRNA gene sequences demonstrated that strain SO07(T) formed a monophyletic group in the gamma-Proteobacteria with only 89 % similarity to members of the genus Halothiobacillus, its nearest phylogenetic neighbours. In addition, the isolate differed from members of the genus Halothiobacillus in its requirement for and tolerance of NaCl; strain SO07(T) was unable to grow in NaCl concentrations of more than 180 mM. On the basis of phylogenetic, chemotaxonomic and physiological data, it is proposed that isolate SO07(T) (=JCM 12417(T) = ATCC BAA-1033(T)) represents the type strain of a novel species in a new genus, Thiovirga sulfuroxydans gen. nov., sp. nov. PMID- 15879234 TI - Alteromonas addita sp. nov. AB - On the basis of phenotypic, genotypic characteristics and analysis of 16S rRNA gene sequences, a novel species belonging to the genus Alteromonas is described. A non-pigmented, motile, Gram-negative bacterium designated R10SW13(T) was isolated from sea water samples collected in Chazhma Bay (Sea of Japan, Pacific Ocean). The novel organism mainly grew between 4 degrees C and 37 degrees C, was neutrophilic and slightly halophilic, tolerating up to 10 % NaCl. Strain R10SW13(T) was haemolytic and was able to degrade starch and Tween 80 and to degrade gelatin and agar weakly, but did not degrade casein. Phosphatidylethanolamine (44.3 +/- 0.9 %) and phosphatidylglycerol (55.7 +/ -0.9 %) were the predominant phospholipids. The major fatty acids formed were typical for the genus Alteromonas, including 16 : 0, 16 : 1omega-7 and 18 : 1omega-7. The G + C content of the DNA was 43.4 mol%. DNA-DNA hybridization experiments showed 38-53 % binding with the DNAs of type strains of phylogenetically related species of the genus Alteromonas, namely: Alteromonas macleodii, Alteromonas marina, Alteromonas stellipolaris, Alteromonas litorea, 'Alteromonas macleodii subsp. fijiensis' and 'Alteromonas infernus'. Based on these results, a novel species, Alteromonas addita sp. nov., is proposed, with strain R10SW13(T) (=KMM 3600(T) = KCTC 12195(T) = LMG 22532(T)) as the type strain. PMID- 15879235 TI - Thioclava pacifica gen. nov., sp. nov., a novel facultatively autotrophic, marine, sulfur-oxidizing bacterium from a near-shore sulfidic hydrothermal area. AB - Strain TL 2(T) was isolated on mineral medium with thiosulfate from a near-shore sulfidic hydrothermal area in Matupi Harbour on the island of New Britain, Papua New Guinea. The cells varied from long filaments with swollen ends, often aggregated, to short rods, depending on the growth conditions. The bacterium was obligately aerobic and grew autotrophically with thiosulfate as energy source or heterotrophically with organic acids and sugars. In thiosulfate-limited continuous culture, mu(max) and Y(max) for autotrophic growth were 0.1 h(-1) and 3 g protein mol(-1), respectively. From the various reduced sulfur compounds tested, only thiosulfate and sulfide supported active respiration. Inorganic carbon was assimilated via the Calvin cycle. Presence of the 'green'-type of form I RubisCO gene was detected. Growth was possible from 15 to 47 degrees C with an optimum at 35 degrees C, pH 6.5-8.5 with an optimum at pH 8.0, and between 10 and 90 g NaCl l(-1) with an optimum at 35 g l(-1). Phylogenetic analysis based on 16S rRNA and cbbL gene sequences demonstrated that strain TL 2(T) forms a separate lineage within the alpha-3 subdivision of the Proteobacteria, distantly related to the genera Rhodovulum and Rhodobacter. On the basis of these results, a novel genus and species, Thioclava pacifica gen. nov., sp. nov., is proposed to accommodate strain TL 2(T) (= DSM 10166(T) = UNIQEM 229(T)). PMID- 15879236 TI - Pusillimonas noertemannii gen. nov., sp. nov., a new member of the family Alcaligenaceae that degrades substituted salicylates. AB - The taxonomic position of a Pseudomonas-like strain, designated BN9(T), was investigated. This strain had previously been isolated as a 5-aminosalicylate degrading organism from a 6-aminonaphthalene-2-sulphonate-degrading mixed bacterial culture. Previously, detection of ubiquinone Q-8, a polyamine pattern with putrescine, spermidine and 2-hydroxyputrescine as the major polyamines, and partial 16S rRNA gene sequencing had suggested that strain BN9(T) belongs to the 'Betaproteobacteria'. This was supported by sequencing the 16S rRNA gene, which demonstrated 94-96 % sequence similarity to different species of the genera Achromobacter, Alcaligenes and Bordetella, and suggested that strain BN9(T) represents a member of the family Alcaligenaceae. Different methods for the construction of phylogenetic dendrograms placed the strain separately from all other genera within the Alcaligenaceae. Fatty acid analysis demonstrated the presence of high concentrations of C(19 : 0) cyclo omega8c. On the basis of low 16S rRNA gene sequence similarity to other members of the Alcaligenaceae, fatty acid and polar lipid profiles, and other unique phenotypic properties of strain BN9(T), the creation of a new genus and species with the name Pusillimonas noertemannii gen. nov., sp. nov. is proposed. The type strain is BN9(T) (= DSM 10065(T) = NCIMB 14020(T)). PMID- 15879237 TI - Pedobacter himalayensis sp. nov., from the Hamta glacier located in the Himalayan mountain ranges of India. AB - Strain HHS 22(T) was isolated from a glacial water sample from the snout of the Hamta glacier located in the Himalayan mountain ranges of India. Phenotypic, chemotaxonomic and phylogenetic analyses established the affiliation of the isolate to the genus Pedobacter. HHS 22(T) exhibits high 16S rRNA gene sequence similarity with Pedobacter cryoconitis (98 %). However, the level of DNA-DNA relatedness between HHS 22(T) and P. cryoconitis is only 42 %. Furthermore, HHS 22(T) differs from P. cryoconitis and the four other recognized species of Pedobacter in a number of phenotypic characteristics. These data suggest that HHS 22(T) represents a novel species of the genus Pedobacter, for which the name Pedobacter himalayensis sp. nov. is proposed. The type strain is HHS 22(T) (= JCM 12171(T) = MTCC 6384(T)). PMID- 15879238 TI - Dinoroseobacter shibae gen. nov., sp. nov., a new aerobic phototrophic bacterium isolated from dinoflagellates. AB - A novel group of aerobic anoxygenic phototrophic bacteria was isolated from marine dinoflagellates, and two strains were characterized in detail. Cells were Gram-negative cocci or ovoid rods and were motile by means of a single, polarly inserted flagellum. They were obligate aerobes requiring 1-7 % salinity. The optimal pH range for growth was 6.5-9.0 and the temperature optimum was 33 degrees C. The bacteria contained bacteriochlorophyll a and spheroidenone as the only carotenoid. The in vivo absorption spectrum displayed two maxima in the infrared region at 804 and 868 nm. The distinct 804 nm band indicates the presence of light-harvesting system 2. Various organic carbon sources were assimilated, including many carboxylic acids, glucose and glycerol, but not butyrate, ethanol or methanol. Dissimilatory nitrate reduction was found for both strains. The physiological characteristics of the new strains resembled those of Roseobacter denitrificans, but there were differences in the lipid composition. Based on 16S rRNA gene sequence analysis the new strains are relatively distant from other recognized species, with the closest relatives Jannaschia helgolandensis, Ruegeria atlantica and Rhodobacter veldkampii showing 94.1-93.4 % similarity. Similarity to Roseobacter denitrificans was only 92.2 %, in line with numerous other species of the Roseobacter group. Therefore, it is proposed to classify the strains in a new genus and species within the Roseobacter clade, Dinoroseobacter shibae gen. nov., sp. nov. The type strain is DFL 12(T) (=DSM 16493(T)=NCIMB 14021(T)). PMID- 15879239 TI - Methanogenium frittonii Harris et al. 1996 is a later synonym of Methanoculleus thermophilus (Rivard and Smith 1982) Maestrojuan et al. 1990. AB - The 16S rRNA gene sequence of [Methanogenium] frittonii DSM 2832(T) was determined and was found to be 99.9 % similar to the sequence of Methanoculleus thermophilus DSM 2373(T). DNA-DNA hybridizations between both strains revealed 86 % DNA-DNA binding, indicating that both strains belong to the same species. The determination of the DNA G + C content of both type strains, DSM 2832(T) and DSM 2373(T), revealed values of 56.1 and 59.1 mol%, respectively. Based on the phenotypic and genotypic characteristics, it is proposed to unite the species [Methanogenium] frittonii and Methanoculleus thermophilus under the name Methanoculleus thermophilus, which is the earlier synonym and hence has priority. Emended descriptions of the species Methanoculleus thermophilus and the genus Methanogenium are also given. PMID- 15879240 TI - Mycobacterium florentinum sp. nov., isolated from humans. AB - Eight mycobacterial strains isolated during an 11 year period from the sputum of independent patients with various pulmonary disorders and, in one case, from a lymph node of a young girl, were found to present identical features. Phenotypic and genotypic characteristics revealed that the most closely related species to these test isolates were Mycobacterium triplex and Mycobacterium lentiflavum. However, the lipids of the cell wall of the test isolates differed from those of the latter species by TLC and presented unique profiles by both GC and HPLC. Genotypic analysis showed that they had unique 16S rRNA gene and internal transcribed spacer (ITS) sequences, and could be differentiated from all other mycobacterial strains by PCR restriction analysis of hsp65. The strains presented high resistance to antimycobacterial drugs. The name Mycobacterium florentinum sp. nov. is proposed for this taxon, with strain FI-93171(T) (=DSM 44852(T) = CIP 108409(T)) as the type strain. PMID- 15879241 TI - Pseudomonas argentinensis sp. nov., a novel yellow pigment-producing bacterial species, isolated from rhizospheric soil in Cordoba, Argentina. AB - During a study in the Argentinian region of Chaco (Cordoba), some strains were isolated from the rhizosphere of grasses growing in semi-desertic arid soils. Two of these strains, one isolated from the rhizospheric soil of Chloris ciliata (strain CH01(T)) and the other from Pappophorum caespitosum (strain PA01), were Gram-negative, strictly aerobic rods, which formed yellow round colonies on nutrient agar. They produced a water-insoluble yellow pigment, and a fluorescent pigment was also detected. A polyphasic taxonomic approach was used to characterize the strains. Comparison of the 16S rRNA gene sequences showed a similarity of 99.3 % between them, and phylogenetic analysis revealed that the strains belong to the genus Pseudomonas, within the gamma-subclass of the Proteobacteria. The closest related species is Pseudomonas straminea IAM 1598(T) (similarity of 99.0 % to strain CH01(T) and 98.8 % to strain PA01), clustering in a separate branch with the various methods of tree building used. Strains CH01(T) and PA01 both had a single polar flagellum, like other yellow pigment-producing pseudomonads related to them. Both strains produced catalase and oxidase. Similar to P. straminea, they did not hydrolyse gelatin or casein. The G+C DNA contents determined were 57.5 mol% for CH01(T) and 58.0 mol% for PA01. DNA-DNA hybridization results showed 81 % relatedness between them, and only 40-44 % relatedness with respect to the type strain of P. straminea. These results, together with other phenotypic characteristics, support the conclusion that both isolates belong to the same species, and should be described as representing a novel species within the genus Pseudomonas, for which the name Pseudomonas argentinensis sp. nov. is proposed. The type strain is CH01(T) (=LMG 22563(T) = CECT 7010(T)). PMID- 15879242 TI - Petrimonas sulfuriphila gen. nov., sp. nov., a mesophilic fermentative bacterium isolated from a biodegraded oil reservoir. AB - A mesophilic, anaerobic, fermentative bacterium, strain BN3(T), was isolated from a producing well of a biodegraded oil reservoir in Canada. Cells were Gram negative, non-motile rods that did not form spores. The temperature range for growth was 15-40 degrees C, with optimum growth at 37-40 degrees C. The strain grew with up 4 % NaCl, with optimum growth in the absence of NaCl. Tryptone was required for growth. Yeast extract and elemental sulfur stimulated growth. Growth was also enhanced during fermentation of glucose, arabinose, galactose, maltose, mannose, rhamnose, lactose, ribose, fructose, sucrose, cellobiose, lactate, mannitol and glycerol. Acetate, hydrogen and CO(2) were produced during glucose fermentation. Elemental sulfur and nitrate were used as electron acceptors and were reduced to sulfide and ammonium, respectively. The G + C content of the genomic DNA was 40.8 mol%. Phylogenetic analyses of the 16S rRNA gene sequence indicated that the strain was a member of the phylum 'Bacteroidetes', distantly related to the genera Bacteroides and Tannerella (similarity values of less than 90 %). The chemotaxonomic data (fatty acids, polar lipids and quinones composition) also indicated that strain BN3(T) could be clearly distinguished from its closest cultivated relatives. This novel organism possesses phenotypic, chemotaxonomic and phylogenetic traits that do not allow its classification as a member of any previously described genus; therefore, it is proposed that this isolate should be described as a member of a novel species of a new genus, Petrimonas gen. nov., of which Petrimonas sulfuriphila sp. nov. is the type species. The type strain is BN3(T) (= DSM 16547(T) = JCM 12565(T)). PMID- 15879243 TI - Bacillus arsenicus sp. nov., an arsenic-resistant bacterium isolated from a siderite concretion in West Bengal, India. AB - Strain Con a/3(T) is a Gram-positive, motile, endospore-forming, rod-shaped and arsenic-resistant bacterium, which was isolated from a concretion of arsenic ore obtained from a bore-hole. The bacterium grew in the presence of 20 mM arsenate and 0.5 mM arsenite. Diaminopimelic acid was present in the cell wall peptidoglycan, MK-7 was the major menaquinone, and iso-C(15 : 0), anteiso-C(15 : 0), iso-C(16 : 0) and C(16 : 1)(delta7cis) were the major fatty acids. Based on its phenotypic, chemotaxonomic and phylogenetic characteristics, strain Con a/3(T) was identified as a member of the genus Bacillus. It exhibited maximum similarity (97 %) at the 16S rRNA gene level with Bacillus barbaricus (DSM 14730(T)); however, the DNA-DNA relatedness value with B. barbaricus was 60 %. Strain Con a/3(T) also exhibited a number of phenotypic differences from B. barbaricus (DSM 14730(T)). Strain Con a/3(T) was therefore identified as representing a novel species of the genus Bacillus, for which the name Bacillus arsenicus sp. nov. is proposed. The type strain is Con a/3(T) (= MTCC 4380(T) = DSM 15822(T) = JCM 12167(T)). PMID- 15879244 TI - Corynebacterium mooreparkense, a later heterotypic synonym of Corynebacterium variabile. AB - Strains of a Gram-positive bacterium were isolated from the Irish smear-ripened cheese Gubbeen, and assigned to a new species, Corynebacterium mooreparkense, in 2001. During a further study on the same cheese, no additional isolates from this species could be found. Instead, multiple isolates of its nearest phylogenetic neighbour, Corynebacterium variabile, were found. A first screening with rep-PCR and SDS-PAGE pointed to a similarity between C. mooreparkense and C. variabile. Following this peculiar result, attempts were made to collect all type strains deposited at different culture collections and all strains described by Brennan et al. [Int J Syst Evol Microbiol (2001) 51, 843-852]. Subsequently, 16S rRNA gene sequencing and DNA-DNA hybridizations were performed. All C. mooreparkense strains had a 16S rRNA gene sequence similarity of at least 99.5 % with C. variabile and the DNA-DNA relatedness was 95 %. On the basis of these results, it is concluded that C. mooreparkense is a later heterotypic synonym of C. variabile. PMID- 15879245 TI - Rhodococcus yunnanensis sp. nov., a mesophilic actinobacterium isolated from forest soil. AB - A Gram-positive, aerobic, non-motile, mesophilic strain, designated YIM 70056(T), was isolated from a forest soil sample in Yunnan Province, China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that this isolate had less than 97.0 % similarity to any Rhodococcus species with validly published names, with the exception of Rhodococcus fascians (DSM 20669(T)), which was found to be its closest neighbour (98.9 % similarity). Chemotaxonomic data, including peptidoglycan type, diagnostic sugar compositions, fatty acid profiles, menaquinones, polar lipids and mycolic acids, were determined for this isolate; the results supported the affiliation of strain YIM 70056(T) to the genus Rhodococcus. The DNA G + C content was 63.5 mol%. The results of DNA-DNA hybridization with R. fascians DSM 20669(T), in combination with chemotaxonomic and physiological data, demonstrated that isolate YIM 70056(T) represents a novel Rhodococcus species, for which the name Rhodococcus yunnanensis sp. nov. is proposed, with YIM 70056(T) (=CCTCC AA 204007(T) = KCTC 19021(T) = DSM 44837(T)) as the type strain. PMID- 15879246 TI - Mycobacterium pseudoshottsii sp. nov., a slowly growing chromogenic species isolated from Chesapeake Bay striped bass (Morone saxatilis). AB - A group of slowly growing photochromogenic mycobacteria was isolated from Chesapeake Bay striped bass (Morone saxatilis) during an epizootic of mycobacteriosis. Growth characteristics, acid-fastness and 16S rRNA gene sequencing results were consistent with those of the genus Mycobacterium. Biochemical reactions, growth characteristics and mycolic acid profiles (HPLC) resembled those of Mycobacterium shottsii, a non-pigmented mycobacterium also isolated during the same epizootic. Sequencing of the 16S rRNA genes, the gene encoding the exported repeated protein (erp) and the gene encoding the 65 kDa heat-shock protein (hsp65) and restriction enzyme analysis of the hsp65 gene demonstrated that this group of isolates is unique. Insertion sequences associated with Mycobacterium ulcerans, IS2404 and IS2606, were detected by PCR. These isolates could be differentiated from other slowly growing pigmented mycobacteria by their inability to grow at 37 degrees C, production of niacin and urease, absence of nitrate reductase, negative Tween 80 hydrolysis and resistance to isoniazid (1 mug ml(-1)), p-nitrobenzoic acid, thiacetazone and thiophene-2 carboxylic hydrazide. On the basis of this polyphasic study, it is proposed that these isolates represent a novel species, Mycobacterium pseudoshottsii sp. nov. The type strain, L15(T), has been deposited in the American Type Culture Collection as ATCC BAA-883(T) and the National Collection of Type Cultures (UK) as NCTC 13318(T). PMID- 15879247 TI - Naxibacter alkalitolerans gen. nov., sp. nov., a novel member of the family 'Oxalobacteraceae' isolated from China. AB - A taxonomic study was performed on strain YIM 31775(T), which was isolated from a soil sample collected from Yunnan Province, China. The isolate was chemo organotrophic, aerobic and Gram-negative. Cells were short rods and motile, with one or more polar flagella. Growth temperature and pH ranged from 4 to 55 degrees C and 6.5 to 12.0, respectively; the optimum growth temperature and pH were 28-37 degrees C and 7.0-9.0, respectively. Q-8 was the predominant respiratory lipoquinone. The major fatty acids were C(16 : 1)omega7c (42.4 %) and C(16 : 0) (28.1 %). The DNA G + C content was 62.4 +/ -0.3 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 31775(T) should be placed within the family 'Oxalobacteraceae', in which it formed a distinct lineage. Based on the high 16S rRNA gene sequence divergence and phenotypic characteristics, it is proposed that strain YIM 31775(T) should be classified as representing a novel member of the family 'Oxalobacteraceae', for which the name Naxibacter alkalitolerans gen. nov., sp. nov. is proposed. The type strain is YIM 31775(T) (= CCTCC AA 204003(T) = KCTC 12194(T)). PMID- 15879248 TI - Lysobacter concretionis sp. nov., isolated from anaerobic granules in an upflow anaerobic sludge blanket reactor. AB - The taxonomic positions of Lysobacter species with validly published names and a novel strain Ko07(T), which was newly isolated from an upflow anaerobic sludge blanket reactor treating wastewater from a brewery, were (re)estimated on the basis of results obtained by using a polyphasic taxonomy approach. Phylogenetic inference based on 16S rRNA gene sequences showed that strain Ko07(T) and all Lysobacter species with validly published names clustered together in a phylogenetic branch within the class 'Gammaproteobacteria'. The sequence similarity of strain Ko07(T) to the type strains of established Lysobacter species was in the range 94.9-96.7 %. Ubiquinone Q-8 and branched fatty acids, C(11 : 0) iso, C(15 : 0) iso, C(16 : 0) iso, iso C(17 : 1)omega9c and C(11 : 0) iso 3OH, predominantly appeared in strain Ko07(T) as well as in all type strains of the recognized Lysobacter species. The DNA-DNA hybridization values of strain Ko07(T) with those of recognized Lysobacter species were estimated to be 2-20 %. Despite sharing common taxonomic features in important phenotypic characteristics, such as gliding movement, long-rod shape and proteolytic activity, strain Ko07(T) could be distinguished from the Lysobacter species with validly published names by its low DNA-DNA hybridization value, a comparatively low DNA G + C content (63.8 mol%), substrate utilization and some physiochemical characteristics. On the basis of the results obtained in this study, it is proposed that strain Ko07(T) should be classified as representing a novel member of the genus Lysobacter, for which the name Lysobacter concretionis sp. nov. is proposed. The type strain is Ko07(T) (=KCTC 12205(T) = DSM 16239(T)). PMID- 15879249 TI - Reclassification of Agrobacterium ferrugineum LMG 128 as Hoeflea marina gen. nov., sp. nov. AB - Members of the species Agrobacterium ferrugineum were isolated from marine environments. The type strain of this species (= LMG 22047(T) = ATCC 25652(T)) was recently reclassified in the new genus Pseudorhodobacter, in the order 'Rhodobacterales' of the class 'Alphaproteobacteria'. Strain LMG 128 (= ATCC 25654) was also initially classified as belonging to the species Agrobacterium ferrugineum; however, the nearly complete 16S rRNA gene sequence of this strain indicated that it does not belong within the genus Agrobacterium or within the genus Pseudorhodobacter. The closest related organism, with 95.5 % 16S rRNA gene similarity, was Aquamicrobium defluvii from the family 'Phyllobacteriaceae' in the order 'Rhizobiales'. The remaining genera from this order had 16S rRNA gene sequence similarities that were lower than 95.1 % with respect to strain LMG 128. These phylogenetic distances suggested that strain LMG 128 belonged to a different genus. The major fatty acid present in strain LMG 128 was mono unsaturated straight chain 18 : 1omega7c. The G + C content of the DNA was 53.1 mol%. Strain LMG 128 grew at 4 degrees C but not at 40 degrees C, and tolerated up to 5 % NaCl. The pH range for growth was 6-8. It produced urease and beta galactosidase, and hydrolysed aesculin. Denitrification was negative. Growth was observed with many carbohydrates as the only carbon source. The data from this polyphasic study indicate that this strain belongs to a new genus of the family 'Phyllobacteriaceae', and therefore it is proposed that strain LMG 128(T) should be reclassified as representing a novel species within the new genus Hoeflea gen. nov., for which the name Hoeflea marina sp. nov. is proposed. PMID- 15879250 TI - Erythrobacter luteolus sp. nov., isolated from a tidal flat of the Yellow Sea in Korea. AB - A Gram-negative, non-spore-forming, yellow-pigmented, slightly halophilic bacterial strain, SW-109(T), was isolated from a tidal flat of the Yellow Sea in Korea, and subjected to a polyphasic taxonomic study. This isolate did not produce bacteriochlorophyll a and contained ubiquinone-10 as the predominant respiratory lipoquinone and C(18 : 1)omega7c as the major fatty acid. The DNA G + C content was 60.3 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain SW-109(T) is phylogenetically affiliated to the genus Erythrobacter of the family Sphingomonadaceae. Strain SW-109(T) exhibited levels of 16S rRNA gene sequence similarity to the type strains of Erythrobacter species of 94.0-96.3 %, making it possible to categorize strain SW-109(T) as a species that is separate from previously recognized Erythrobacter species. On the basis of its phenotypic properties and phylogenetic distinctiveness, SW-109(T) (= KCTC 12311(T) = JCM 12599(T)) was classified as the type strain of a novel Erythrobacter species, for which the name Erythrobacter luteolus sp. nov. is proposed. PMID- 15879251 TI - Application of a recN sequence similarity analysis to the identification of species within the bacterial genus Geobacillus. AB - Full-length recN and 16S rRNA gene sequences were determined for a collection of 68 strains from the thermophilic Gram-positive genus Geobacillus, members of which have been isolated from geographically and ecologically diverse locations. Phylogenetic treeing methods clustered the isolates into nine sequence similarity groups, regardless of which gene was used for analysis. Several of these groups corresponded unambiguously to known Geobacillus species, whereas others contained two or more type strains from species with validly published names, highlighting a need for a re-assessment of the taxonomy for this genus. For taxonomic analysis of bacteria related at a genus, species or subspecies level, recN sequence comparisons had a resolving power nearly an order or magnitude greater than 16S rRNA gene comparisons. Mutational saturation rendered recN comparisons much less powerful than 16S rRNA gene comparisons for analysis of higher taxa, however. Analysis of recN sequences should prove a powerful tool for assigning strains to species within Geobacillus, and perhaps within other genera as well. PMID- 15879252 TI - Alcanivorax dieselolei sp. nov., a novel alkane-degrading bacterium isolated from sea water and deep-sea sediment. AB - Two bacterial strains, B-5(T) and NO1A, were isolated from the surface water of the Bohai Sea and deep-sea sediment of the east Pacific Ocean, respectively. Both strains were halophilic, aerobic, Gram-negative, non-spore-forming, catalase- and oxidase-positive motile rods. They grew on a restricted spectrum of organic compounds, including some organic acids and alkanes. On the basis of 16S rRNA gene sequence similarity, strains B-5(T) and NO1A were shown to belong to the gamma-Proteobacteria. Highest similarity values were found with Alcanivorax venustensis (95.2 %), Alcanivorax jadensis (94.6 %) and Alcanivorax borkumensis (94.1 %). Principal fatty acids of both strains were C(16 : 0), C(16 : 1)omega7c and C(18 : 1)omega7c. The chemotaxonomically characteristic fatty acid C(19 : 0) cyclo omega8c was also detected. On the basis of the above, together with results of physiological and biochemical tests, DNA-DNA hybridization, comparisons of 16S 23S internal transcribed spacer sequences and comparisons of the partial deduced amino acid sequence of alkane hydroxylase, both strains were affiliated to the genus Alcanivorax but were differentiated from recognized Alcanivorax species. Therefore, a novel species, Alcanivorax dieselolei sp. nov., represented by strains B-5(T) and NO1A is proposed, with the type strain B-5(T) (=DSM 16502(T)=CGMCC 1.3690(T)). PMID- 15879253 TI - Burkholderia phytofirmans sp. nov., a novel plant-associated bacterium with plant beneficial properties. AB - A Gram-negative, non-sporulating, rod-shaped, motile bacterium, with a single polar flagellum, designated strain PsJN(T), was isolated from surface-sterilized onion roots. This isolate proved to be a highly effective plant-beneficial bacterium, and was able to establish rhizosphere and endophytic populations associated with various plants. Seven related strains were recovered from Dutch soils. Based on 16S rRNA gene sequence data, strain PsJN(T) and the Dutch strains were identified as representing a member of the genus Burkholderia, as they were closely related to Burkholderia fungorum (98.7 %) and Burkholderia phenazinium (98.5 %). Analysis of whole-cell protein profiles and DNA-DNA hybridization experiments confirmed that all eight strains belonged to a single species. Strain PsJN(T) had a DNA G+C content of 61.0 mol%. Only low levels of DNA-DNA hybridization to closely related species were found. Qualitative and quantitative differences in fatty acid composition between strain PsJN(T) and closely related species were identified. The predominant fatty acids in strain PsJN(T) were 16 : 0, 18 : 1omega7c and summed feature 3 (comprising 16 : 1omega7c and/or iso-15 : 0 2-OH). Isolate PsJN(T) showed high 1-aminocyclopropane-1-carboxylate deaminase activity and is therefore able to lower the ethylene level in a developing or stressed plant. Production of the quorum-sensing signal compound 3-hydroxy-C8 homoserine lactone was detected. Based on the results of this polyphasic taxonomic study, strain PsJN(T) and the seven Dutch isolates are considered to represent a single, novel species, for which the name Burkholderia phytofirmans sp. nov. is proposed. The type strain is strain PsJN(T) (=LMG 22146(T) = CCUG 49060(T)). PMID- 15879254 TI - Clostridium sardiniense Prevot 1938 and Clostridium absonum Nakamura et al. 1973 are heterotypic synonyms: evidence from phylogenetic analyses of phospholipase C and 16S rRNA sequences, and DNA relatedness. AB - Clostridium sardiniense Prevot 1938 and Clostridium absonum Nakamura et al. 1973 have long been considered similar in terms of their biological and biochemical properties, but their taxonomic positions have not been clarified by DNA-DNA hybridization studies or rigorous analysis of 16S rRNA genes. In the present study, DNA-DNA hybridization analysis revealed that C. absonum strains DSM 599(T), DSM 600 and KZ 1544 shared 83.0-86.3 % DNA relatedness with C. sardiniense DSM 2632(T). 16S rRNA gene sequence analysis showed that the C. absonum strains also shared high identity with C. sardiniense DSM 2632(T) (99.7, 99.3 and 99.8 % for DSM 599(T), DSM 600 and KZ 1544, respectively), implying that C. absonum and C. sardiniense are synonyms. In addition, alignment of the inferred amino acid sequences for phospholipase C (PLC) indicated 96.5 % identity between PLCs from C. sardiniense and C. absonum, but relatively low identity with other clostridial species. These results strongly suggest that the species C. sardiniense and C. absonum should be united, with the name C. sardiniense having priority. PMID- 15879255 TI - Mucispirillum schaedleri gen. nov., sp. nov., a spiral-shaped bacterium colonizing the mucus layer of the gastrointestinal tract of laboratory rodents. AB - The mammalian gastrointestinal tract is covered by a layer of mucus that can harbour a range of bacterial species specifically adapted to colonize this ecological niche. Examination of 110 bacterial isolates cultivated from the gastrointestinal tract of 23 mice revealed the presence of a subgroup of 30 isolates that did not correspond genetically with genera commonly associated with this site, i.e. members of the epsilon-Proteobacteria such as Helicobacter and Campylobacter species. Instead this group of isolates was found to lie within the phylum Deferribacteres, a completely distinct lineage in the domain Bacteria. There was a high level of consensus in results obtained from the phenotypic and genotypic characterization of a number of the isolates, which showed they were distinct from other members of the Deferribacteres. As such, they are proposed to constitute a new genus and species, Mucispirillum schaedleri gen. nov., sp. nov. These organisms are anaerobic, Gram-negative, spiral-shaped rods with bipolar flagella. The type strain is HRI I17(T) (= ATCC BAA-1009(T) = ACM 5223(T)). PMID- 15879256 TI - Comparative study of overlapping genes in bacteria, with special reference to Rickettsia prowazekii and Rickettsia conorii. AB - Overlapping genes have been proposed as a means of achieving genome reduction by compressing the maximum amount of information in limited sequence space. In this report, comparative analyses of the overlapping genes of genomes of nine bacteria with different lifestyles were performed. The results clearly suggest that overlapping genes may be a result of evolutionary pressure to minimize genome size. The genomes of two closely related obligatory intracellular parasites - Rickettsia prowazekii and Rickettsia conorii - were investigated further. Detailed analyses of these two genomes revealed that mutations at the ends of coding regions and elimination of intergenic DNA are the main forces that determine overlapping of genes. PMID- 15879257 TI - Bacillus alveayuensis sp. nov., a thermophilic bacterium isolated from deep-sea sediments of the Ayu Trough. AB - Two thermophilic, spore-forming strains, TM1(T) and TM5, were isolated from deep sea sediment (4000 m below sea level) of the Ayu Trough in the western Pacific Ocean. Cells of the two strains were Gram-positive, motile and rod-shaped. Their spores were ellipsoidal, subterminal to terminal and occurred in swollen sporangia. The two strains grew at temperatures up to 65 degrees C and in the pH range 6.5-9.0. The NaCl concentration for optimal growth was 3.0 % (w/v) and growth was inhibited by 5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains TM1(T) and TM5 belonged to the genus Bacillus, and that strain TM1(T) was most closely related to Bacillus aeolius DSM 15084(T) (96.7 %), Bacillus smithii DSM 4216(T) (96.1 %), Bacillus methanolicus NCIMB 13113(T) (95.8 %) and Bacillus pallidus DSM 3670(T) (95.7 %). Between the 16S rRNA gene sequences of strains TM1(T) and TM5 there were only three nucleotide differences, implying that the two strains were of the same species. The cellular fatty acid profiles of the two strains were also very similar, with iso-C(15 : 0), iso-C(16 : 0), C(16 : 0), iso-C(17 : 0) and anteiso-C(17 : 0) as the major components. The G + C content of strain TM1(T) was 38.7 %. On the basis of phenotypic and molecular data, strains TM1(T) and TM5 represent a novel species of the genus Bacillus, for which the name Bacillus alveayuensis sp. nov. is proposed. The type strain is TM1(T) (= KCTC 10634(T) = JCM 12523(T)). PMID- 15879258 TI - Marinitoga hydrogenitolerans sp. nov., a novel member of the order Thermotogales isolated from a black smoker chimney on the Mid-Atlantic Ridge. AB - A novel, thermophilic, anaerobic bacterium that is able to tolerate hydrogen was isolated from a deep-sea hydrothermal chimney collected at the Rainbow field on the Mid-Atlantic Ridge. Cells were rod-shaped and surrounded by a sheath-like outer structure (toga); they were weakly motile by means of a polar flagellum. They appeared singly, in pairs or in short chains. They grew at 35-65 degrees C (optimum 60 degrees C), pH 4.5-8.5 (optimum pH 6.0) and 10-65 g sea salts l(-1) (optimum 30-40 g l(-1)). The isolate was organotrophic, and able to grow on various carbohydrates or complex proteinaceous substrates. Growth was not inhibited under 100 % hydrogen or in the presence of 2 % oxygen in the gas phase. The isolate reduces sulfur, although sulfur reduction is not required for growth. The fermentation products identified on glucose were acetate, ethanol, formate, hydrogen and CO(2). The G + C content of the genomic DNA was 28 +/- 1 mol%. Phylogenetic analysis of the 16S rRNA gene placed the strain within the genus Marinitoga, order Thermotogales, in the bacterial domain. On the basis of the 16S rRNA gene sequence comparisons and physiological characteristics, the isolate is considered to represent a novel species, for which the name Marinitoga hydrogenitolerans sp. nov. is proposed. The type strain is AT1271(T) (=DSM 16785(T) = JCM 12826(T)). PMID- 15879259 TI - Sphingopyxis baekryungensis sp. nov., an orange-pigmented bacterium isolated from sea water of the Yellow Sea in Korea. AB - A Gram-negative, motile, slightly halophilic bacterial strain, SW-150(T), was isolated from sea water of the Yellow Sea, Korea, and was characterized by a polyphasic taxonomic approach. Strain SW-150(T) grew optimally at 25-30 degrees C and in the presence of 2 % (w/v) NaCl. The isolate could be distinguished from other Sphingopyxis species in producing an orange pigment. It contained ubiquinone-10 as the predominant respiratory lipoquinone and C(18 : 1)omega7c and C(17 : 1)omega6c as the major fatty acids. No 3-hydroxy fatty acids were detected. Major polar lipids were sphingoglycolipid, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 63 mol%. Comparative 16S rRNA gene sequence analyses showed that strain SW-150(T) was phylogenetically affiliated to the genus Sphingopyxis of the family Sphingomonadaceae. Similarity values between the 16S rRNA gene sequences of strain SW-150(T) and the type strains of Sphingopyxis species ranged from 91.6 to 94.2 %, making it possible to categorize strain SW-150(T) as a species that is separate from previously described Sphingopyxis species. On the basis of phenotypic properties and phylogenetic distinctiveness, SW-150(T) (=KCTC 12231(T) = DSM 16222(T)) should be classified as the type strain of a novel Sphingopyxis species, for which the name Sphingopyxis baekryungensis sp. nov. is proposed. PMID- 15879260 TI - Novosphingobium taihuense sp. nov., a novel aromatic-compound-degrading bacterium isolated from Taihu Lake, China. AB - A novel aromatic-compound-degrading bacterium, strain T3-B9(T), was isolated from sediment of Taihu Lake, Jiangsu Province, south-east China. This bacterial isolate assimilated several aromatic compounds such as phenol, aniline, nitrobenzene, 4-chloronitrobenzene and phenanthrene. The taxonomy of strain T3 B9(T) was studied by polyphasic methods. The organism showed a range of phenotypic and chemotaxonomic properties consistent with those of the genus Novosphingobium. The 16S rRNA gene sequence similarity of strain T3-B9(T) to members of the genus Novosphingobium ranged from 91.6 to 97.5 %, and this isolate clustered phylogenetically with members of genus Novosphingobium. The DNA-DNA relatedness values of strain T3-B9(T) to the most phylogenetically related species, Novosphingobium subterraneum DSM 12447(T), Novosphingobium aromaticivorans ATCC 700278(T) and Novosphingobium stygium ATCC 700280(T), were 31, 33 and 14 %, respectively. The combined genotypic and phenotypic data show that strain T3-B9(T) represents a novel species of the genus Novosphingobium, for which the name Novosphingobium taihuense sp. nov. is proposed. The type strain is T3-B9(T) (=AS 1.3432(T) = JCM 12465(T)). PMID- 15879261 TI - Pleomorphomonas oryzae gen. nov., sp. nov., a nitrogen-fixing bacterium isolated from paddy soil of Oryza sativa. AB - Five strains of free-living diazotrophs isolated from rice were characterized by using a polyphasic approach. The strains were found to be very closely related, with 99-100 % 16S rRNA gene sequence similarity and DNA-DNA hybridization values greater than 70 %, suggesting that they represent a single species. When compared with other recognized species, they showed not more than 93 and 89 % similarity for the 16S rRNA and nifH gene sequences, respectively. Phylogenetic distances showed that these isolates were distinct from other taxa within the alpha Proteobacteria. Chemotaxonomic characteristics of these isolates included the DNA G + C content (62.1-63.1 mol%), the major quinone system (Q-10), predominant fatty acids (18 : 1omega7c, cyclo 19 : 0omega8c and 16 : 0) and major hydroxy fatty acids (14 : 0 3-OH, 18 : 0 3-OH and 16 : 0 3-OH). Based on phylogenetic and phenotypic analyses, these isolates are considered to represent a novel genus and species, for which the name Pleomorphomonas oryzae gen. nov., sp. nov. is proposed. The type strain is F-7(T) (=IAM 15079(T) = ATCC BAA-940(T) = DSM 16300(T)). PMID- 15879262 TI - Asticcacaulis taihuensis sp. nov., a novel stalked bacterium isolated from Taihu Lake, China. AB - A novel stalked bacterium, designated strain T3-B7(T), was isolated from sediment of Taihu Lake, Jiangsu Province, China, and its taxonomy was studied by using a polyphasic approach. Cell morphology, physiological and biochemical properties, and polar lipids indicated that strain T3-B7(T) represented a member of the genus Asticcacaulis. Based on 16S rRNA gene sequence similarity analysis, strain T3 B7(T) was found to be phylogenetically related to Asticcacaulis biprosthecium DSM 4723(T) (98.5 %) and Asticcacaulis excentricus DSM 4724(T) (95.0 %), but could be differentiated from these two species on the basis of the number and position of prosthecae, assimilation of sugars, nitrate reduction and tolerance to NaCl. Levels of DNA-DNA relatedness of strain T3-B7(T) to A. biprosthecium DSM 4723(T) and A. excentricus DSM 4724(T) were 37.1 and 18.0 %, respectively. The G + C content of strain T3-B7(T) was 59 mol% (T(m)). It is concluded that strain T3 B7(T) represents a novel species of the genus Asticcacaulis, for which the name of Asticcacaulis taihuensis sp. nov. is proposed. The type strain is T3-B7(T) (=AS 1.3431(T) = JCM 12463(T)). PMID- 15879263 TI - Slackia faecicanis sp. nov., isolated from canine faeces. AB - Morphological, biochemical and molecular genetic studies were carried out on an unknown non-spore-forming, Gram-positive, rod-shaped bacterium that was isolated from dog faeces. The bacterium grew under strictly anaerobic conditions, was asaccharolytic, and possessed a relatively high G + C content of 61 mol%. Phylogenetic analysis based on comparative 16S rRNA gene sequencing showed that the unidentified bacterium was a member of the family Coriobacteriaceae and represents a hitherto unknown subline within the genus Slackia. Based on the presented findings, a novel species, Slackia faecicanis sp. nov., is described. The type strain of Slackia faecicanis is 5WC12(T) (= CCUG 48399(T) = CIP 108281(T)). PMID- 15879264 TI - Labrys methylaminiphilus sp. nov., a novel facultatively methylotrophic bacterium from a freshwater lake sediment. AB - A new bacterial isolate from a methylamine enrichment culture is described, representing a novel species of facultatively methylotrophic bacteria. The non motile bacterium is Gram-negative, replicates by budding and does not form endospores. The isolate utilizes methylated amines, as well as a variety of monosaccharides, disaccharides, amino acids, organic acids, aromatic compounds and alcohols as substrates, but does not utilize methanol. Growth factors are not required, although yeast extract stimulates growth. The major components of the fatty acid profile are C(18 : 1)omega7c, C(19 : 0) cyclo and C(16 : 0). The dominant cellular phospholipids are phosphatidyl acid, phosphatidylcholine and phosphatidylethanolamine. The G + C content of the DNA is 65.7 +/- 0.3 mol%. 16S rRNA gene-based phylogenetic analysis revealed that the novel isolate belongs to the alpha-Proteobacteria and is closely related to the only representative of the genus Labrys, Labrys monachus (97.4 % sequence similarity). However, the level of DNA-DNA relatedness with L. monachus is less than 3 %, justifying the placement of this isolate into a novel species of the genus Labrys. The name Labrys methylaminiphilus sp. nov. is proposed (type strain JLW10(T) = ATCC BAA-1080(T) = DSM 16812(T)). PMID- 15879265 TI - Dechloromonas denitrificans sp. nov., Flavobacterium denitrificans sp. nov., Paenibacillus anaericanus sp. nov. and Paenibacillus terrae strain MH72, N2O producing bacteria isolated from the gut of the earthworm Aporrectodea caliginosa. AB - Earthworms emit nitrous oxide (N(2)O) via the activity of bacteria in their gut. Four N(2)O-producing facultative aerobes, ED1(T), ED5(T), MH21(T) and MH72, were isolated from the gut of the earthworm Aporrectodea caliginosa. The isolates produced N(2)O under conditions that simulated the microenvironment of the earthworm gut. ED1(T) and ED5(T) were Gram-negative, motile rods that carried out complete denitrification (i.e. the reduction of nitrate to N(2)) and contained membranous c-type cytochromes. ED1(T) grew optimally at 30 degrees C and pH 7. ED1(T) oxidized organic acids and reduced (per)chlorate, sulfate, nitrate and nitrite. The closest phylogenetic relative of ED1(T) was Dechloromonas agitata. ED5(T) grew optimally at 25 degrees C and pH 7. ED5(T) grew mainly on sugars, and nitrate and nitrite were used as alternative electron acceptors. The closest phylogenetic relatives of ED5(T) were Flavobacterium johnsoniae and Flavobacterium flevense. MH21(T) and MH72 were motile, spore-forming, rod-shaped bacteria with a three-layered cell wall. Sugars supported the growth of MH21(T) and MH72. Cells of MH21(T) grew in chains, were linked by connecting filaments and contained membranous b-type cytochromes. MH21(T) grew optimally at 30-35 degrees C and pH 7.7, grew by fermentation and reduced low amounts of nitrite to N(2)O. The closest phylogenetic relatives of MH21(T) were Paenibacillus borealis and Paenibacillus chibensis. Based on morphological, physiological and phylogenetic characteristics, ED1(T) (= DSM 15892(T) = ATCC BAA-841(T)), ED5(T) (= DSM 15936(T) = ATCC BAA-842(T)) and MH21(T) (=DSM 15890(T) = ATCC BAA-844(T)) are proposed as type strains of the novel species Dechloromonas denitrificans sp. nov., Flavobacterium denitrificans sp. nov. and Paenibacillus anaericanus sp. nov., respectively. MH72 is considered a new strain of Paenibacillus terrae. PMID- 15879266 TI - Leuconostoc durionis sp. nov., a heterofermenter with no detectable gas production from glucose. AB - Three lactic acid bacterial (LAB) strains obtained from a Malaysian acid fermented condiment, tempoyak (made from pulp of the durian fruit), showed analogous but distinct patterns after screening by SDS-PAGE of whole-cell proteins and comparison with profiles of all recognized LAB species. 16S rRNA gene sequencing of one representative strain showed that the taxon belongs phylogenetically to the genus Leuconostoc, with its nearest neighbour being Leuconostoc fructosum (98 % sequence similarity). Biochemical characteristics and DNA-DNA hybridization experiments demonstrated that the strains differ from Leuconostoc fructosum and represent a single, novel Leuconostoc species for which the name Leuconostoc durionis sp. nov. is proposed. The type strain is LMG 22556(T) (= LAB 1679(T) = D-24(T) = CCUG 49949(T)). PMID- 15879267 TI - Arcobacter halophilus sp. nov., the first obligate halophile in the genus Arcobacter. AB - A Gram-negative bacterium, designated LA31B(T), was isolated from water collected from a hypersaline lagoon on Laysan Atoll in the north-western Hawaiian Islands. Single cells of LA31B(T) were slightly curved but became helical as their length increased. Preliminary characterization based on 16S rRNA gene sequence analysis showed that LA31B(T) shared 96.0 % identity with an Arcobacter sp. isolated from a cyanobacterial mat in hypersaline Lake Sinai, and 94 % identity with Arcobacter nitrofigilis, the type species of the genus Arcobacter. A polyphasic taxonomic study was conducted and confirmed the phylogenetic affiliation of strain LA31B(T) to the genus Arcobacter. However, LA31B(T) was found to be distinct from all recognized Arcobacter species, by a comprehensive biochemical test analysis, whole-cell fatty acid profiling, DNA G + C content (35 mol% in LA31B(T)) and degree of DNA-DNA reassociation. Most notably, LA31B(T) was found to be an obligate halophile, a hitherto undescribed feature among recognized Arcobacter species. These data indicate that LA31B(T) should be considered to represent a novel species in the genus Arcobacter, for which the name Arcobacter halophilus sp. nov. is proposed. This is the first obligately halophilic member of the genus. The type strain is LA31B(T) (=ATCC BAA-1022(T) = CIP 108450(T)). PMID- 15879268 TI - Bacillus axarquiensis sp. nov. and Bacillus malacitensis sp. nov., isolated from river-mouth sediments in southern Spain. AB - Two Gram-positive, rod-shaped, endospore-forming bacteria (strains CR-119(T) and CR-95(T)) were isolated from brackish sediments in the mouth of the river Velez in Malaga, southern Spain, and subjected to a polyphasic taxonomic study. Phenotypic tests showed that these strains were related to other Bacillus species at a similarity level of less than 87.6 %. Both strains are halotolerant, aerobic, chemoheterotrophic, motile with peritrichous flagella and biosurfactant producers. Their endospores are oval, subterminal and non-deforming structures. The predominant menaquinone in both strains is MK-7. The fatty-acid profiles of both strains contain large quantities of branched and saturated fatty acids. The major fatty acids (%) are 15 : 0 anteiso (32.4), 15 : 0 iso (16.8), 17 : 0 iso (13.4), 16 : 0 (11.5) and 17 : 0 anteiso (10.2) in strain CR-119(T) and 15 : 0 anteiso (37.5), 17 : 0 iso (16.0) and 17 : 0 anteiso (15.8) in strain CR-95(T). The G + C contents of strains CR-119(T) and CR-95(T) are 41.0 and 42.5 mol%, respectively. RAPD analysis confirmed the low degree of similarity between the two strains and also amongst other Bacillus species. 16S rRNA gene analysis of strain CR-119(T) showed the highest sequence similarity to be 97.4 %, with Bacillus mojavensis and Bacillus subtilis subsp. spizizenii. In the case of strain CR-95(T), the maximum similarity value was 99.5 %, with B. mojavensis. DNA DNA hybridization of strains CR-119(T) and CR-95(T) with the above species produced values lower than 46.9 %. Therefore, on the basis of phenotypic characteristics, phylogenetic data and genomic distinctiveness, we conclude that these Bacillus strains merit classification as novel species, for which we propose the names Bacillus axarquiensis sp. nov. (type strain CR-119(T) = CECT 5688(T) = LMG 22476(T)) and Bacillus malacitensis sp. nov. (type strain CR-95(T) = CECT 5687(T) = LMG 22477(T)). PMID- 15879269 TI - Transfer of Chryseobacterium meningosepticum and Chryseobacterium miricola to Elizabethkingia gen. nov. as Elizabethkingia meningoseptica comb. nov. and Elizabethkingia miricola comb. nov. AB - The taxonomic positions of six strains (including the type strain) of Chryseobacterium meningosepticum (King 1959) Vandamme et al. 1994 and the type strain of Chryseobacterium miricola Li et al. 2004 were re-evaluated by using a polyphasic taxonomic approach. Phylogenetic analysis, based on 16S rRNA gene sequencing, showed that the strains represent a separate lineage from the type strains of the Chryseobacterium-Bergeyella-Riemerella branch within the family Flavobacteriaceae (90.7-93.9 % similarities), which was supported by phenotypic differences. Combined phylogenetic and phenotypic data showed that C. meningosepticum and C. miricola should be transferred to a new genus, Elizabethkingia gen. nov., with the names Elizabethkingia meningoseptica comb. nov. (type strain, ATCC 13253(T) = NCTC 10016(T) = LMG 12279(T) = CCUG 214(T)) and Elizabethkingia miricola comb. nov. (type strain, DSM 14571(T) = JCM 11413(T) = GTC 862(T)) proposed. PMID- 15879270 TI - Dyadobacter crusticola sp. nov., from biological soil crusts in the Colorado Plateau, USA, and an emended description of the genus Dyadobacter Chelius and Triplett 2000. AB - Bacterial strain CP183-8(T) was isolated from biological soil crusts collected in the Colorado Plateau, USA. Cells of this strain were aerobic, non-motile, Gram negative, psychrotolerant and formed beaded chains in the stationary growth phase. They contained C(16 : 1)omega5c and C(16 : 1)omega7c as major fatty acids. 16S rRNA gene sequence analysis assigned the strain to the genus Dyadobacter. However, it shared a sequence similarity of only 95.88 % with the type strain of Dyadobacter fermentans, NS114(T). Because it also exhibited a significant number of phenotypic and chemotaxonomic differences from D. fermentans, it is described as a novel second species in the genus Dyadobacter, with the name Dyadobacter crusticola sp. nov. The type strain is CP183-8(T) (=DSM 16708(T) = ATCC BAA 1036(T)). PMID- 15879271 TI - Chryseobacterium taichungense sp. nov., isolated from contaminated soil. AB - A bacterial strain (CC-TWGS1-8(T)) isolated from a tar-contaminated soil in Taiwan was studied in a detailed taxonomic study. The cells were Gram-negative, rod-shaped and non-spore-forming. Phylogenetic analyses using the 16S rRNA gene sequence of the strain clearly revealed an affiliation to the genus Chryseobacterium, the highest sequence similarities being to the type strain of Chryseobacterium indologenes (96.8 %), to Chryseobacterium gleum (96.8 %) and to Chryseobacterium joostei (96.4 %). The 16S rRNA gene sequence similarities to all other Chryseobacterium species were below 96 %. The major whole-cell fatty acids were 15 : 0 iso (35.4 %) and 17 : 0 iso 3OH (22.5 %). DNA-DNA hybridization values and the biochemical and chemotaxonomic properties demonstrate that strain CC-TWGS1-8(T) represents a novel species, for which the name Chryseobacterium taichungense sp. nov. is proposed. The type strain is CC-TWGS1-8(T) (= CCUG 50001(T) = CIP 108519(T)). PMID- 15879272 TI - Paenibacillus rhizosphaerae sp. nov., isolated from the rhizosphere of Cicer arietinum. AB - Two sporulating bacterial strains designated CECAP06(T) and CECAP16 were isolated from the rhizosphere of the legume Cicer arietinum in Argentina. Almost-complete 16S rRNA gene sequences identified the isolates as a Paenibacillus species. It was most closely related to Paenibacillus cineris LMG 18439(T) (99.6 % sequence similarity), Paenibacillus favisporus LMG 20987(T) (99.4 % sequence similarity) and Paenibacillus azoreducens DSM 13822(T) (97.7 % sequence similarity). The cells of this novel species were motile, sporulating, rod-shaped, Gram-positive and strictly aerobic. The predominant fatty acids were anteiso-C(15 : 0), C(16 : 0) and iso-C(16 : 0). The DNA G+C content of strains CECAP06(T) and CECAP16 was 51.3 and 50.9 mol%, respectively. Growth was observed from many carbohydrates, but gas production was not observed from glucose. Catalase and oxidase activities were present. The isolates produced beta-galactosidase and hydrolysed aesculin. Gelatinase, caseinase and urease were not produced. The results of DNA-DNA hybridization showed that the strains from this study constitute a novel species of the genus Paenibacillus, for which the name Paenibacillus rhizosphaerae sp. nov. is proposed. The type strain is CECAP06(T) (=LMG 21955(T) = CECT 5831(T)). PMID- 15879273 TI - Natrinema altunense sp. nov., an extremely halophilic archaeon isolated from a salt lake in Altun Mountain in Xinjiang, China. AB - A novel extremely halophilic strain, AJ2(T), was isolated from Ayakekum salt lake located in the Altun Mountain National Nature Reserve in Xinjiang, China. This isolate was neutrophilic, motile and grew in a wide range of MgCl(2) concentrations (0.005-1.0 M). The major polar lipids of the isolate were C(20)C(20) and C(20)C(25) derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate. A comprehensive 16S rRNA gene sequence analysis revealed that the isolate shared 96.6-97.7 % sequence identity with Natrinema species. The isolate, however, could be genetically differentiated from these species by DNA-DNA hybridization analysis and on the basis of its physiological properties. On the basis of the polyphasic evidence, strain AJ2(T) (= AS 1.3731(T) = JCM 12890(T)) represents the type strain of a novel species, for which the name Natrinema altunense sp. nov. is proposed. PMID- 15879274 TI - Pedobacter caeni sp. nov., a novel species isolated from a nitrifying inoculum. AB - A Gram-negative, rod-shaped, non-spore-forming bacterium was isolated from a nitrifying inoculum. On the basis of 16S rRNA gene sequence similarity, LMG 22862(T) was shown to belong to the family Sphingobacteriaceae, related to Pedobacter africanus (98.0 %) and Pedobacter heparinus (97.6 %). The results of 16S rRNA gene sequence analysis, DNA-DNA hybridization, SDS-PAGE, analysis of the fatty acid composition and physiological and biochemical tests allowed genotypic and phenotypic differentiation of LMG 22862(T) from Pedobacter species with validly published names. LMG 22862(T) therefore represents a novel species within this genus, for which the name Pedobacter caeni sp. nov. is proposed, with the type strain LMG 22862(T) (=DSM 16990(T)). PMID- 15879275 TI - Syntrophobacter sulfatireducens sp. nov., a novel syntrophic, propionate oxidizing bacterium isolated from UASB reactors. AB - Two obligate anaerobes, TB8106(T) and WZH410, which degraded propionate in syntrophic association with methanogens, were isolated from two upflow anaerobic sludge blanket reactors, one treating brewery wastewater and the other bean curd wastewater. The strains were Gram-negative, non-spore-forming and non-motile. Cells were egg-shaped, with a size of 1.0-1.3 x 1.8-2.2 microm. Growth was observed at 20-48 degrees C and pH 6.2-8.8. Both strains converted propionate to acetate and methane in co-culture with methanogens, and grew on propionate plus sulfate in pure culture, with a doubling time of 52-55 h at 37 degrees C. Sulfate and thiosulfate both served as electron acceptors for propionate degradation. The DNA G + C contents of the two strains were 58.5 and 58.7 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains were closely related to a propionate-oxidizing syntrophic bacterium, Syntrophobacter fumaroxidans DSM 10017(T) (94.7 % similarity). However, the novel strains could not ferment fumarate, and grew at a more alkaline pH range than Syntrophobacter fumaroxidans. Moreover, the novel isolates had obviously higher growth rates on propionate plus sulfate (0.12 day(-1)) than Syntrophobacter fumaroxidans DSM 10017(T) (0.024 day(-1)). Therefore, a novel species, Syntrophobacter sulfatireducens sp. nov., is proposed, with strain TB8106(T) (=AS 1.5016(T) = DSM 16706(T)) as the type strain. PMID- 15879276 TI - Reclassification of Pediococcus urinaeequi (ex Mees 1934) Garvie 1988 as Aerococcus urinaeequi comb. nov. AB - The taxonomic status of Pediococcus urinaeequi is described, and the transfer of the species to the genus Aerococcus with the name Aerococcus urinaeequi comb. nov. is proposed, on the basis of the analysis of 16S rRNA gene sequence and DNA DNA hybridization data. PMID- 15879277 TI - Streptomyces sodiiphilus sp. nov., a novel alkaliphilic actinomycete. AB - An alkaliphilic actinomycete, strain YIM 80305(T), which was isolated from a muddy sample in Chaka salt lake, Qinghai Province of China, was characterized using a polyphasic approach. The isolate produced light-yellow substrate and yellow-white aerial mycelia on most tested media. Optimum pH for growth was 9.0 10.0 with scant growth at pH 7.0. Results showed that strain YIM 80305(T) was obligately Na(+)-dependent, and showed sensitivity to K(+). The DNA G + C content was 70.5 mol%. 16S rRNA gene sequence analysis together with these characteristics consistently assigned strain YIM 80305(T) to the genus Streptomyces. It formed a distinct clade based on analyses of the almost-complete and 120-nucleotide variable gamma region of the 16S rRNA gene. It could be differentiated by phenotypic and genotypic analysis from all the Streptomyces species whose names have been validly published. On the basis of polyphasic evidence, Streptomyces sodiiphilus sp. nov. is proposed. The type strain is YIM 80305(T) (= CCTCC AA 203015(T) = CIP 107975(T)). PMID- 15879278 TI - Bacteroides massiliensis sp. nov., isolated from blood culture of a newborn. AB - Gram-negative anaerobic rods were isolated from blood culture of a healthy baby born at term. Based on phylogenetic analysis and specific phenotypic characteristics, this strain was included within the Bacteroides cluster. The novel bile-resistant anaerobic species was designated Bacteroides massiliensis and it has 92.8 % 16S rRNA similarity to Bacteroides vulgatus and a DNA G + C content of 49 mol%. The major cellular fatty acid is anteiso-branched C(15 : 0). The type strain of B. massiliensis sp. nov. is B84634(T) (=CIP 107942(T) = CCUG 48901(T)). PMID- 15879279 TI - Lentibacillus salarius sp. nov., isolated from saline sediment in China, and emended description of the genus Lentibacillus. AB - A Gram-positive, spore-forming and moderately halophilic bacterium, strain BH139(T), was isolated from saline sediment of Xinjiang Province, China. Cells of strain BH139(T) were motile, flagellated rods. The organism grew optimally at 30 35 degrees C in the presence 12-14 % (w/v) NaCl. The major fatty acids were branched saturated fatty acids such as iso-C(16 : 0), anteiso-C(15 : 0), iso-C(15 : 0), iso-C(14 : 0) and anteiso-C(17 : 0). The G + C content of the genomic DNA was about 43 mol% and the predominant isoprenoid quinone was MK-7. Comparative 16S rRNA gene sequence analyses showed that strain BH139(T) was most closely related to Lentibacillus salicampi KCCM 41560(T) (96.9 % 16S rRNA gene sequence similarity) and formed a distinct phyletic line from that species. On the basis of physiological and molecular properties, the isolate represents a novel species within the genus Lentibacillus, for which the name Lentibacillus salarius sp. nov. is proposed. The type strain is BH139(T) (= KCTC 3911(T) = DSM 16459(T)). PMID- 15879280 TI - Reclassification of Nocardia corynebacterioides Serrano et al. 1972 (Approved Lists 1980) as Rhodococcus corynebacterioides comb. nov. AB - The type strain of Nocardia corynebacterioides was the subject of a polyphasic taxonomic study. The 16S rRNA gene sequence was aligned with the sequences of representatives of the genera Corynebacterium, Dietzia, Gordonia, Mycobacterium, Nocardia, Rhodococcus, Skermania, Tsukamurella and Williamsia, and phylogenetic trees were constructed by using maximum-parsimony, maximum-likelihood and neighbour-joining methods. It was evident from the phylogenetic analysis that N. corynebacterioides represents a distinct phyletic line within the genus Rhodococcus. Menaquinone analysis showed that the organism contained dihydrogenated menaquinone with eight isoprene units, MK-8(H(2)), as the major isoprenologue. The genealogical evidence, together with chemotaxonomic and phenotypic data from this and previous studies, indicates that N. corynebacterioides DSM 20151(T) (= CIP 104510(T)) should be reclassified in the genus Rhodococcus as Rhodococcus corynebacterioides comb. nov. PMID- 15879281 TI - Marinobacter hydrocarbonoclasticus Gauthier et al. 1992 and Marinobacter aquaeolei Nguyen et al. 1999 are heterotypic synonyms. AB - The genus Marinobacter was described with a single species, Marinobacter hydrocarbonoclasticus Gauthier et al. 1992, to place a hydrocarbon-degrading marine bacterium belonging to the gamma-subclass of the Proteobacteria. Marinobacter aquaeolei Nguyen et al. 1999, the second described species of the genus, was proposed for a strain isolated from an oil-producing well on an offshore platform in southern Vietnam, on the basis of differences in the whole cell protein pattern and lipopolysaccharide composition, although its phenotypic and genotypic characteristics were very similar to those of the type species, M. hydrocarbonoclasticus. In the present paper, literature data concerning the two species were reviewed. Fatty acid composition, G+C content and DNA-DNA hybridization studies were performed in order to clarify the taxonomic positions of these two species. Based on the results obtained in this study and phenotypic and phylogenetic traits available from the literature, it is proposed that the two species should be united under the same name; according to the rules of priority, M. hydrocarbonoclasticus, the first described species, is the earlier synonym and M. aquaeolei is the later synonym. PMID- 15879282 TI - Taxonomy and phylogeny of the ascomycetous yeast genus Zygoascus, with proposal of Zygoascus meyerae sp. nov. and related anamorphic varieties. AB - Physiological characters, mating compatibility, PCR-RAPD fingerprints, mol% G + C content, DNA-DNA relatedness, and large-subunit and internal transcribed spacer rRNA gene sequences of strains assigned to the genus Zygoascus were re-examined. On the basis of those data, and after phylogenetic analyses, an emendation of Zygoascus hellenicus (type material is a cross of CBS 6736(T) x CBS 5839(T)) is proposed, comprising two novel anamorphic varieties, Candida steatolytica var. steatolytica (CBS 6736(T)) and C. steatolytica var. inositophila (CBS 5839(T)). A novel teleomorphic species, Zygoascus meyerae sp. nov. (type material is a cross of CBS 4099(T) x CBS 7521(T)) is described, together with two novel anamorphic varieties corresponding to it, Candida hellenica var. hellenica (CBS 4099(T)) and C. hellenica var. acidophila (CBS 7115(T)). PMID- 15879283 TI - Cryptococcus taeanensis sp. nov., a new anamorphic basidiomycetous yeast isolated from a salt farm. AB - Cryptococcus taeanensis, a new anamorphic yeast species originating from a salt farm on the Taean peninsula in Korea, is described. Strain 3-12(T) grew by budding, contained ubiquinone Q-10 and xylose in cell hydrolysates, utilized d glucuronate and did not ferment D-glucose. A molecular phylogenetic analysis based on the large-subunit rRNA D1/D2 domain and ITS region sequences placed C. taeanensis near Auriculibuller fuscus and Bullera japonica, recently proposed taxa of the Tremellales. However, these species were distinguishable based on standard physiological tests used for yeast identification, with characteristics including the assimilation of L-sorbose, absence of ballistoconidia, no arbutin hydrolysis and no growth in the presence of 0.01 % cycloheximide. The isolate exhibited the typical physiology of the genus Cryptococcus Vuillemin, but its large-subunit rRNA D1/D2 domain sequence was clearly distinct from previously described species in the genus. Therefore, on the basis of these results, Cryptococcus taeanensis sp. nov. is proposed; the type strain is 3-12(T) (=KCTC 17149(T) = CBS 9742(T)). PMID- 15879284 TI - Metschnikowia hamakuensis sp. nov., Metschnikowia kamakouana sp. nov. and Metschnikowia mauinuiana sp. nov., three endemic yeasts from Hawaiian nitidulid beetles. AB - Three heterothallic, haplontic yeast species, Metschnikowia hamakuensis, Metschnikowia kamakouana and Metschnikowia mauinuiana, are described from isolates associated with endemic nitidulid beetles living on various endemic plants on three Hawaiian islands. As morphospecies, they are similar to Metschnikowia hawaiiensis, but based on mating compatibility and ascospore formation, they can be assigned clearly to distinct biological species. Analysis of ITS/5.8S and D1/D2 large subunit rDNA sequences shows that, with M. hawaiiensis and two other isolates, these species form a distinct subclade within the large-spored Metschnikowia species, indicating that they are Hawaiian endemics. Type cultures are: M. hamakuensis, UWOPS 04-207.1(T) = CBS 10056(T) = NRRL Y-27834(T) (type, h(+)) and UWOPS 04-204.1 = CBS 10055 = NRRL Y-27833 (allotype, h(-)); M. kamakouana, UWOPS 04-112.5(T) = CBS 10058(T) = NRRL Y 27836(T) (type, h(+)) and UWOPS 04-109.1 = CBS 10057 = NRRL Y-27835 (allotype, h( )); and M. mauinuiana, UWOPS 04-190.1(T) = CBS 10060(T) = NRRL Y-27838(T) (type, h(+)) and UWOPS 04-110.4 = CBS 10059 = NRRL Y-27837 (allotype, h(-)). PMID- 15879285 TI - Pichia myanmarensis sp. nov., a novel cation-tolerant yeast isolated from palm sugar in Myanmar. AB - Four halotolerant yeast strains, M21(T), M34-1, HS054 and D41, were isolated from various foods in South-East Asia. These isolates were most closely related to Pichia anomala, with which each strain had from zero to two differences in the 26S rDNA D1/D2 domain nucleotide sequence; for this reason, they were thought to be the same as, or sister species of, P. anomala. Of the four yeast isolates, only one strain, M21(T), had an 18S rDNA sequence that differed from those of P. anomala IFO 10213(T) and the other three isolates, having 20 substitutions and two gaps. Strain M21(T) showed lower cation (Li(+)) tolerance (< or =0.3 M LiCl) than P. anomala IFO 10213(T) or the other three strains (< or = 0.5 M LiCl). Furthermore, the DNA-DNA hybridization data indicated that M21(T) was clearly distinct from P. anomala IFO 10213(T) and the other three isolates. The ability of strain M21(T) to assimilate d-arabinose distinguished it from P. anomala IFO 10213(T) and the other three isolates; it also differed in that it was able to grow at 37 and 40 degrees C. Strain M21(T) grew by multilateral budding, produced persistent asci, in which between one and four hat-shaped ascospores were formed, and contained ubiquinone Q-7. On the basis of this polyphasic characterization, strain M21(T) represents a novel species within the Q-7-containing group of the genus Pichia, for which the name Pichia myanmarensis is proposed. The type strain is M21(T) (= NBRC 11090(T) = JCM 12922(T) = CBS 9786(T)). PMID- 15879286 TI - Phylogenetic analysis and description of Eperythrozoon coccoides, proposal to transfer to the genus Mycoplasma as Mycoplasma coccoides comb. nov. and Request for an Opinion. AB - Eperythrozoon coccoides, an epierythrocytic organism that causes a mild haemolytic anaemia in laboratory and wild mice, currently is thought to be a rickettsia. To determine the relationship of this agent to other haemotrophic bacterial parasites, the 16S rRNA gene of this organism has been sequenced and it is shown by phylogenetic analysis that this wall-less bacterium is not a rickettsia but actually is a mycoplasma. This mycoplasma shares properties with and is closely related to the other uncultivated mycoplasmas that comprise a recently identified group, the haemotrophic mycoplasmas (haemoplasmas). The haemoplasma group is composed of former Eperythrozoon and Haemobartonella species as well as newly identified haemotrophic mycoplasmas. Haemoplasmas parasitize the surface of erythrocytes of a wide variety of vertebrate animal hosts and are transmitted mainly by blood-feeding arthropod vectors. Because both primary infections and chronic latent infections caused by this bacterium have been observed in many laboratories and this bacterium has been the subject of much experimental work, considerable information exists about this haemotrophic mycoplasma that may be applicable to other haemoplasmas. It is proposed that Eperythrozoon coccoides be reclassified as Mycoplasma coccoides comb. nov. A Request for an Opinion is submitted to the Judicial Commission of the International Committee on Systematics of Prokaryotes regarding this reclassification. PMID- 15879287 TI - Erythropoietin improves functional and histological outcome in neonatal stroke. AB - Neonatal stroke is a condition that leads to disability in later life, and as yet there is no effective treatment. Recently, erythropoietin (EPO) has been shown to be cytoprotective following brain injury and may promote neurogenesis. However, the effect of EPO on functional outcome and on morphologic changes in neonatal subventricular zone (SVZ) following experimental neonatal stroke has not been described. We used a transient focal model of neonatal stroke in P10 rat. Injury was documented by diffusion weighted MRI during occlusion. Immediately upon reperfusion, either EPO (5U/gm) or vehicle was administered intraperitoneally and animals were allowed to grow for 2 wk. Sensorimotor function was assessed using the cylinder rearing test and then brains were processed for volumetric analysis of the SVZ. Stroke induced SVZ expansion proportional to hemispheric volume loss. EPO treatment markedly preserved hemispheric volume and decreased the expansion of SVZ unilaterally. Furthermore, EPO treatment significantly improved the asymmetry of forelimb use following neonatal stroke. This functional improvement directly correlated with the amount of preserved hemispheric volume. These results suggest EPO may be a candidate in the treatment of neonatal stroke. PMID- 15879288 TI - Changes in fetal lung distension alter expression of vascular endothelial growth factor and its isoforms in developing rat lung. AB - Vascular endothelial growth factor A (VEGF-A) is essential for normal pulmonary vascular and parenchymal development. Changes in fetal lung distension profoundly affect lung growth and maturation, including vascular development. To define developmental lung expression of VEGF-A and its receptors and investigate effects of changes in fetal lung distension, we studied fetal rats at embryonic day (ED) 16, 19, and 22, postnatal rats at postnatal day (PD) 5, 10, and 21, and adult rats. We used reverse transcriptase PCR to measure mRNA expression for VEGF-A isoforms (VEGF-A(120), (-144), (-164), and (-188)) and VEGF-A receptors, Flt-1 and Flk-1. With advancing development, mRNA content increased only for VEGF A(188) (p < 0.05) and for Flt-1 (p < 0.02) and Flk-1 (p < 0.005). As a percentage of total VEGF-A mRNA, VEGF-A(188) (15% at ED 16) increased to become the dominant isoform at PD 21 (40%, p < 0.005) and adulthood; in contrast, there were decreases in both VEGF-A(144) (p < 0.05) and (-120) (p < 0.005). VEGF-A protein was expressed in alveolar epithelium (type I and II cells) and interstitium. Increasing fetal lung distension by tracheal occlusion (TO) accelerated the normal maturational pattern of VEGF-A isoforms and increased VEGF-A protein; decreasing fetal lung distension by congenital diaphragmatic hernia (CDH) retarded the normal developmental pattern and decreased VEGF-A protein. Neither TO nor CDH consistently affected Flt-1 or Flk-1 mRNA content. These results show that mechanical factors significantly affect lung VEGF-A expression and suggest that VEGF-A mediates previously described changes in lung vascular and parenchymal development caused by CDH and by TO. PMID- 15879289 TI - Lipopolysaccharide induces both a primary and a secondary phase of sensitization in the developing rat brain. AB - Data indicate that bacterial products in combination with other antenatal or postnatal exposures increase the risk of perinatal brain injury. We have previously shown that administration of lipopolysaccharide (LPS) 4 h before hypoxia-ischemia (HI) increases brain injury in 7-d-old rats. The mechanisms behind such sensitization are unclear, but contrasts against a preconditioning effect of LPS given 1-3 d before ischemia in adult animals. To investigate how the effects of LPS depend on the time interval between administration and HI in the developing brain, we evaluated the effect of varying time interval (2-72 h) between LPS and HI, the duration of HI (20 or 50 min), and age of the rat pups (postnatal d 4 or 7). Outcome was assessed by brain injury scoring of specific regions. We found that LPS reduced brain injury (by 78%) when administered 24 h before 50 min of HI. However, when LPS was administered 6 h before either 20 or 50 min of HI, brain injury was increased by 2026% and 137%, respectively. Even LPS given 72 h before HI increased injury, both when LPS was administered at postnatal d 4 (by 446%) and 7 (by 77%). In conclusion, LPS enhanced vulnerability in the developing brain both in the acute (4-6 h) and the chronic (72 h) phase after administration, whereas an intermediate interval between LPS and HI had the opposite effect. The long-term sensitizing effect of LPS has not been previously described. PMID- 15879290 TI - Monocyte toll-like receptor 4 expression and LPS-induced cytokine production increase during gestational aging. AB - Premature newborns are highly susceptible to severe bacterial infections. This is partially due to their immature innate immune system, characterized by decreased neutrophil and monocyte activity as well as by reduced concentrations of complement factors. However, additional mechanisms might be important for innate immunity and are still the subject of considerable debate. The importance of pattern recognition domains such as Toll-like receptors (TLR) has been fully acknowledged within the last few years. Therefore, we investigated age-related monocyte TLR4 expression and lipopolysaccharide-induced cytokine secretion from very low birth weight infants (VLBWI) and from newborns after wk 30 of gestation in comparison to healthy adults. In VLBWI, expression of TLR4 surface protein, detected by flow cytometry, and TLR4-specific mRNA, quantified by real time-PCR, were significantly reduced in comparison to mature infants and to adults. Reduced TLR4 expression was paralleled by significantly diminished ex vivo LPS stimulated IL-1beta, IL-6, and tumor necrosis factor-alpha secretion into whole blood. We conclude that, in VLBWI, the minimized expression of TLR4 contributes to the susceptibility of VLBWI to infections with Gram-negative bacteria due to the lack of cytokines to boost initial immune response. PMID- 15879291 TI - Higher spontaneous and TNFalpha-induced apoptosis of neonatal blood granulocytes. AB - Granulocytes play an important role in inflammatory diseases. Neonates tend to develop granulocytopenia under sepsis and stress. It remains unclear whether apoptosis of neonatal granulocytes is different from that of adult granulocytes. In this study, we analyzed the discrepancy of granulocyte apoptosis between cord blood (CB) and adult blood (AB). We found that spontaneous and tumor necrosis factor-alpha (TNFalpha)-induced granulocyte apoptosis as determined by phosphatidylserine expression and DNA fragmentation were more prominent in CB granulocytes than AB granulocytes. CD95 ligand and TNFalpha levels were significantly higher in CB than in AB (p = 0.001 and p < 0.001, respectively). TNF receptor-2 and CD95 expression on CB granulocytes were not different from those on AB granulocytes. However, the TNF receptor-1 (TNFR1) expression was lower on CB granulocytes than that on AB granulocytes (69.98 +/- 7.32 versus 89.04 +/- 3.73%, p = 0.029). This decrease of TNFR1 expression on neonatal granulocytes might be related to a higher plasma TNFalpha level associated with an intrinsic defect on the dynamic change of membrane TNFR1 expression in neonatal granulocytes. The expression of anti-apoptotic molecule Bcl-2 in neonatal granulocytes was also lower than that in adult granulocytes (4.64 +/- 0.51 versus 7.24 +/- 1.17 unit/mg protein, p = 0.032). Moreover, another anti apoptotic signal, nuclear factor-kappaB nuclear translocation, was also lower in CB than AB granulocytes. Results from this study suggest that higher plasma death ligands associated with lower anti-apoptotic molecules in granulocytes may act an important role in triggering neonatal granulocyte apoptosis. PMID- 15879292 TI - Effects of systemic and local phenylephrine and arginine vasopressin infusions in conscious postnatal sheep. AB - Mean arterial pressure (MAP) increases after birth, however, the mechanisms remain unclear. Systemic angiotensin II (ANG II) infusions increase MAP in newborn sheep, but the direct effects of ANG II on peripheral vascular resistance (PVR) are minimal. Thus, its systemic pressor effects may reflect release of other pressor agents, e.g. alpha-agonists and/or AVP, suggesting they contribute to postnatal regulation of MAP and PVR. To address this, we performed studies in conscious sheep at 7-14, 15-21, and 22-35 d postnatal, infusing phenylephrine (PE) or AVP systemically or intra-arterially into the hindlimb while measuring MAP, heart rate (HR), and femoral blood flow (FmBF). Basal MAP and FmBF rose, whereas HR and femoral vascular resistance (FmVR) fell (p < or = 0.03) during the first month postnatal. Although systemic PE and AVP dose dependently increased MAP and FmVR and decreased FmBF and HR (p < 0.001, ANOVA) at all ages, responses were not age dependent. Notably, increases in FmVR exceeded increases in MAP, and responses to PE appeared to exceed AVP (p < 0.05). Hindlimb infusions of both agents decreased FmBF and increased FmVR dose dependently (p < 0.001, ANOVA) at all ages without altering MAP or HR. These responses also were not age dependent. Unlike ANG II, PE and AVP directly increase PVR in newborn sheep. Moreover, FmVR increases more than MAP at all doses, suggesting these agonists may contribute to postnatal MAP regulation and could mediate the effects of systemic ANG II on postnatal MAP. PMID- 15879293 TI - Effects of erythropoietin on hyperoxic lung injury in neonatal rats. AB - Pulmonary oxygen toxicity is believed to play a prominent role in the lung injury that leads to the development of bronchopulmonary dysplasia (BPD). To determine whether human recombinant erythropoietin (rhEPO) treatment reduces the risk of developing BPD, we investigated the effect of rhEPO treatment on the histopathologic changes seen in hyperoxia-induced lung injury of BPD. Twenty-five rat pups were divided into four groups: air-exposed control group (n = 5), hyperoxia-exposed placebo group (n = 7), hyperoxia-exposed rhEPO-treated group (n = 6), and air-exposed rhEPO-treated group (n = 7). Measurement of alveolar surface area, quantification of secondary crest formation, microvessel count, evaluation of alveolar septal fibrosis, and smooth muscle actin immunostaining were performed to assess hyperoxia-induced changes in lung morphology. Treatment of hyperoxia-exposed animals with rhEPO resulted in a significant increase in the mean alveolar area, number of secondary crests formed, and the microvessel count in comparison with hyperoxia-exposed placebo-treated animals. There was significantly less fibrosis in rhEPO-treated animals. However, treatment of hyperoxia-exposed animals with rhEPO did not result in a significant change in smooth muscle content compared with hyperoxia-exposed placebo treated animals. Our results suggest treatment with rhEPO during hyperoxia exposure is associated with improved alveolar structure, enhanced vascularity, and decreased fibrosis. Therefore, we conclude that treatment of preterm infants with EPO might reduce the risk of developing BPD. PMID- 15879294 TI - Developmental changes in HIF transcription factor in carotid body: relevance for O2 sensing by chemoreceptors. AB - Before birth, the peripheral chemoreceptors located in the carotid bodies (CB) are adapted to the low fetal Po(2) and are relatively insensitive to hypoxia. After birth, the sensitivity of the CB to hypoxia is reset in response to the rise in Po(2). The mechanism underlying this resetting, which requires several days to complete, remains unknown. We have investigated the possibility that the hypoxia-inducible factors HIF-1alpha and HIF-2alpha, which are activated by oxygen deprivation, are involved in this resetting process. Accordingly, we used immunostaining and densitometry to quantitate the levels of the HIF-1alpha and HIF-2alpha proteins in the rat CB during early perinatal life and after exposure to in vivo hypoxia during adolescence. Tyrosine hydroxylase (TH) was used as a marker for catecholaminergic neurons and oxygen-sensitive cells in the CB. Double immunostaining revealed constitutive expression of HIF-1alpha in both glomus cells (TH+) and sustentacular cells (TH-) of the CB of adolescent rats. However, immunoreactivity toward HIF-2alpha was restricted to glomus cells. After exposure to hypoxia (8% O(2), 6 h), the expression of HIF-1alpha was selectively up regulated in glomus cells and apparent translocation of both HIF-1alpha and HIF 2alpha to the nucleus was observed. Both of these proteins were expressed constitutively in the CB during the perinatal transition period. During the first postnatal week, the intensity of immunostaining for HIF-1alpha in glomus cells decreased markedly, whereas the level of HIF-2alpha remained constant. We suggest that this selective down-regulation of HIF-1alpha may be involved in the postnatal maturation of CB responsiveness to hypoxia. PMID- 15879295 TI - White matter injury following prolonged free radical formation in the 0.65 gestation fetal sheep brain. AB - Free radicals seem to be involved in the development of cerebral white matter damage after asphyxia in the premature infant. The immature brain may be at increased risk of free radical mediated injury, as particularly the preterm infant has a relative deficiency in brain antioxidants systems, such as superoxide dismutase and glutathione peroxidase. In vitro studies show that immature oligodendrocytes express an intrinsic vulnerability to reactive oxygen species and free radical scavengers are able to protect immature oligodendrocytes from injury. The aim of this study was to examine the formation of ascorbyl radicals as a marker of oxidative stress in the preterm brain in association with cerebral white matter injury after intrauterine asphyxia. Fetal sheep at 0.65 gestation were chronically instrumented with vascular catheters and an occluder cuff around the umbilical cord. A microdialysis probe was placed in the periventricular white matter. Fetal asphyxia was induced by occlusion of the umbilical cord for 25 min (n = 10). Microdialysis samples were collected for 72 h and analyzed for ascorbyl radicals using electron spin resonance. Five instrumented fetuses served as controls. Three days after the insult, fetal brains were examined for morphologic injury. Umbilical cord occlusion resulted in prolonged and marked increase in ascorbyl radical production in the brain in connection with white matter injury, with activation of microglia cells in periventricular white matter and axonal injury. These data suggest that reperfusion injury following asphyxia in the immature brain is associated with marked free radical production. PMID- 15879296 TI - Relations of body fatness and cardiovascular fitness to lipid profile in black and white adolescents. AB - High body fatness is associated with unfavorable cardiovascular disease risk profiles in juveniles. However, the degree to which the deleterious effects attributed to fatness may actually be due to the low cardiovascular fitness (CVF) that is usually confounded with fatness is not known. This study determined in 14 to 18-y-old (n = 398) youths the degree to which percentage body fat (%BF) and CVF explained variability in lipids and lipoproteins. Blood samples were taken after a 12-h fast. %BF was measured with dual-energy x-ray absorptiometry. CVF was determined with a multistage treadmill test. The degree to which %BF and CVF explained variance in outcome variables was determined by regression analyses, controlling for demographic variables before entering %BF or CVF and their interactions with race and sex. Because %BF and CVF were highly correlated (r = 0.69, p < 0.001), they were first entered separately in the regression models before being considered together. Both higher %BF and lower CVF were associated with unfavorable concentrations of total cholesterol to HDLC ratio and triglycerides. However, the effects of %BF and CVF were not additive; once %BF was in the regression model, CVF did not explain additional variance. For Lp(a), only %BF explained a significant portion of the variance. For triglycerides, there was a %BF by race interaction, such that the deleterious effects of high %BF were greater in whites than in blacks. These results suggest that interventions to improve lipid profile in youths should be designed primarily to minimize fatness. PMID- 15879297 TI - Inhaled nitric oxide enhances distal lung growth after exposure to hyperoxia in neonatal rats. AB - Exposure of newborn rats to hyperoxia impairs alveolarization and vessel growth, causing abnormal lung structure that persists during infancy. Recent studies have shown that impaired angiogenesis due to inhibition of vascular endothelial growth factor (VEGF) signaling decreases alveolar and vessel growth in the developing lung, and that nitric oxide (NO) mediates VEGF-dependent angiogenesis. The purpose of this study was to determine whether hyperoxia causes sustained reduction of lung VEGF, VEGF receptor, or endothelial NO synthase (eNOS) expression during recovery, and whether inhaled NO improves lung structure in infant rats after neonatal exposure to hyperoxia. Newborn rat pups were randomized to hyperoxia [fraction of inspired oxygen (Fio(2)), 1.00] or room air exposure for 6 d, and then placed in room air with or without inhaled NO (10 ppm) for 2 wk. Rats were then killed for studies, which included measurements of body weight, lung weight, right ventricular hypertrophy (RVH), morphometric analysis of alveolarization (by mean linear intercept (MLI), radial alveolar counts (RAC), and vascular volume (Vv), and immunostaining and Western blot analysis. In comparison with controls, neonatal hyperoxia reduced body weight, increased MLI, and reduced RAC in infant rats. Lung VEGF, VEGFR-2, and eNOS protein expression were reduced after hyperoxia. Inhaled NO treatment after hyperoxia increased body weight and improved distal lung growth, as demonstrated by increased RAC and Vv and decreased MLI. We conclude that neonatal hyperoxia reduced lung VEGF expression, which persisted during recovery in room air, and that inhaled NO restored distal lung growth in infant rats after neonatal hyperoxia. PMID- 15879298 TI - CC10 administration to premature infants: in search of the "silver bullet" to prevent lung inflammation. PMID- 15879299 TI - High expression of pulmonary proteinase-activated receptor 2 in acute and chronic lung injury in preterm infants. AB - Proteinase-activated receptor 2 (PAR(2)), a G-protein-coupled receptor activated by serine proteinases such as trypsin, has been suggested to play an important role in inflammatory and fibroproliferative processes. In preterm infants, the development of bronchopulmonary dysplasia (BPD) is characterized by early pulmonary inflammation and subsequent interstitial fibrosis. High pulmonary trypsin-2 has been shown to be associated with the development of BPD. We studied the expression and distribution of PAR(2) and trypsin-2 by immunohistochemistry in autopsy lung specimens of fetuses (n = 10), of preterm infants who died of acute or prolonged respiratory distress syndrome (RDS) (n = 8 and n = 7, respectively) or BPD (n = 6), and of newborn infants without lung disease (n = 5) who served as controls. In prolonged RDS and BPD, PAR(2) immunoreactivity was significantly higher in bronchial epithelium when compared with infants without pulmonary pathology (p < 0.05 and p < 0.005, respectively). In alveolar epithelium, expression of PAR(2) was elevated in prolonged RDS when compared with newborn infants without pulmonary pathology (p < 0.05). Moreover, strong expression of PAR(2) was detected in myofibroblasts of thickened and fibrotic alveolar walls in prolonged RDS or BPD. Trypsin-2 was co-localized with PAR(2) in bronchoalveolar epithelium. These findings suggest that PAR(2), possibly activated by trypsin-2, may participate in inflammation and fibroproliferation associated with progression of RDS toward BPD in preterm infants. PMID- 15879300 TI - Thymidine phosphorylase gene transfer inhibits vascular smooth muscle cell proliferation by upregulating heme oxygenase-1 and p27KIP1. AB - OBJECTIVE: Thymidine phosphorylase (TP) reportedly promotes endothelial cell migration and induces heme oxygenase (HO)-1 expression. However, its effect on vascular smooth muscle cells (VSMCs) is poorly understood. In this study, we examined the effect of TP on VSMCs in vitro and in vivo. METHODS AND RESULTS: Phagemid vector encoding human TP gene was transfected into rat VSMCs, and a clone overexpressing TP was selected (C2). C2 showed a slower migration and proliferation than VSMCs cloned with empty vector (pC) under basal, serum stimulated, and hypoxic conditions. This decrease in proliferation correlated with TP-induced HO-1 expression and was reversed by inhibitors of either TP or HO activity. Furthermore, in C2, the cyclin-dependent kinase inhibitor (p27KIP1) was much more abundant than in pC, and the cell cycle was arrested at the G1 phase. TP or HO activity inhibitors decreased p27(KIP1) expression in C2 to the level seen in pC. Adventitial TP gene delivery significantly reduced neointimal VSMC migration and neointima formation in balloon-injured rat carotid arteries. CONCLUSIONS: TP overexpression upregulated HO-1 expression and consequently increased p27(KIP1) in cultured VSMCs, and inhibited VSMC migration and proliferation in vitro and in vivo. TP represents a promising target for treating vascular obstructive disease. PMID- 15879301 TI - Sphingomyelinase induces aggregation and fusion of small very low-density lipoprotein and intermediate-density lipoprotein particles and increases their retention to human arterial proteoglycans. AB - OBJECTIVE: Infiltration of low-density lipoprotein (LDL) into subendothelial space is an early step in atherosclerosis. In addition to LDL particles, small very low-density lipoprotein (sVLDL) and intermediate-density lipoprotein (IDL) particles are also able to enter the arterial intima and be retained within the subendothelial extracellular matrix. Here we compared how proteolysis with alpha chymotrypsin and phospholipid hydrolysis with phospholipase A2 or sphingomyelinase (SMase) of sVLDL, IDL, and LDL particles can influence their aggregation, fusion, and binding to human arterial proteoglycans in vitro. METHODS AND RESULTS: In each of the 3 lipoprotein classes, the particles became only slightly aggregated with alpha-chymotrypsin or phospholipase A2. However, the particles strongly aggregated when treated with SMase. The aggregated/fused particles were found to bind to proteoglycans in proteoglycan affinity chromatography more tightly than the native-sized counterparts. In addition, in a microtiter well assay, the binding of SMase-treated lipoproteins was enhanced: the amounts of proteoglycan-bound SMase-treated LDL, IDL, and sVLDL were 4-, 5-, and 20-fold higher, respectively, than the amounts of proteoglycan-bound native lipoproteins. CONCLUSIONS: These results imply a specific role for SMase as an sVLDL- and IDL-modifying enzyme and also suggest a novel mechanism of lipid accumulation in atherogenesis, namely enhanced retention of atherogenic triglyceride-rich lipoprotein particles in intimal areas expressing extracellular SMase activity. PMID- 15879302 TI - Sensitivity and specificity of the ankle-brachial index to predict future cardiovascular outcomes: a systematic review. AB - OBJECTIVE: The ankle-brachial index is the ratio of the ankle and the brachial systolic blood pressure and is used to assess individuals with peripheral arterial disease. An ankle-brachial index <0.90 suggests the presence of peripheral arterial disease and is a marker of cardiovascular risk. The objective of this review is to determine the sensitivity and specificity of an ankle brachial index <0.90 to predict future cardiovascular events, including coronary heart disease, stroke, and death. METHODS AND RESULTS: We conducted a systematic review of the literature and included studies that used an ankle-brachial index cutoff between 0.80 and 0.90 to classify patients with or without peripheral arterial disease, followed patients prospectively, and recorded cardiovascular outcomes (ie, myocardial infarction, stroke, or mortality). Data were combined using a random-effects model meta-analysis to determine the sensitivity, specificity, relative risks, and likelihood ratios of a low ankle-brachial index to predict future cardiovascular disease. A total of 22 studies were identified, 13 were excluded, and 9 studies were included in the meta-analysis. The sensitivity and specificity of a low ankle-brachial index to predict incident coronary heart diseases were 16.5% and 92.7%, for incident stroke were 16.0% and 92.2%, and for cardiovascular mortality were 41.0% and 87.9%, respectively. The corresponding positive likelihood ratios were 2.53 (95% CI, 1.45 to 4.40) for coronary heart disease, 2.45 (95% CI, 1.76 to 3.41) for stroke, and 5.61 (95% CI, 3.45 to 9.13) for cardiovascular death. CONCLUSIONS: The specificity of a low ankle-brachial index to predict future cardiovascular outcomes is high, but its sensitivity is low. The ankle-brachial index should become part of the vascular risk assessment among selected individuals. PMID- 15879303 TI - Genetic determinants of cardiovascular disease risk in familial hypercholesterolemia. AB - OBJECTIVE: To investigate the contribution of polymorphisms in multiple candidate genes to cardiovascular disease (CVD) risk in a large cohort of patients with heterozygous familial hypercholesterolemia (FH). METHODS AND RESULTS: We genotyped 1940 FH patients for 65 polymorphisms in 36 candidate genes. During 91.451 person-years, 643 (33.1%) patients had at least 1 cardiovascular event. Multifactorial Cox survival analysis revealed that the G20210A polymorphism in the prothrombin gene was strongly associated with a significantly increased CVD risk (GA versus GG; P<0.001). CONCLUSIONS: In a large cohort of FH patients, we found that the G20210A polymorphism in the prothrombin gene is strongly associated with CVD risk. Our results constitute a step forward in the unraveling of the hereditary propensity toward CVD in FH and might lead to better risk stratification and hence to more tailored therapy for CVD prevention. PMID- 15879304 TI - A Novel alphavbeta3 integrin antagonist suppresses neointima formation for more than 4 weeks after balloon injury in rats. AB - OBJECTIVES: We performed a detailed kinetic analysis in a rat balloon injury model to clarify the essential roles of alphavbeta3 integrin and endothelial cell (EC) regeneration in neointima formation. Using this model, we evaluated the antistenotic effect of Dainippon compound BS-1417, a novel alphavbeta3 integrin antagonist. METHODS AND RESULTS: Kinetic analysis using RT-PCR showed that alphavbeta3 integrin-related genes are upregulated before neointima formation. Morphological and functional analyses revealed that EC regeneration requires >4 weeks after injury, and that recovery of EC normal function coincides with the arrest of neointima formation. Subcutaneous infusion of BS-1417 for 2, 4, 7, or 12 weeks after injury potently inhibited neointima formation without affecting EC regeneration. Although withdrawal of treatment with BS-1417 after short-term administration after injury resulted in catch-up growth of neointima, a long-term study suggested that this catch-up growth can be prevented by continuous administration of BS-1417 until EC regeneration. CONCLUSIONS: We clarified that alphavbeta3 integrin and EC regeneration play an essential role in neointima formation, and that continuous administration of BS-1417 potently and stably inhibits neointima formation without affecting EC regeneration. These findings suggest that BS-1417 might be useful as a novel systemic drug for the treatment of restenosis. PMID- 15879305 TI - Vascular consequences of endothelial nitric oxide synthase uncoupling for the activity and expression of the soluble guanylyl cyclase and the cGMP-dependent protein kinase. AB - Endothelial dysfunction in the setting of cardiovascular risk factors, such as hypercholesterolemia, hypertension, diabetes mellitus, chronic smoking, as well as in the setting of heart failure, has been shown to be at least partly dependent on the production of reactive oxygen species (ROS), such as the superoxide radical, and the subsequent decrease in vascular bioavailability of nitric oxide (NO). Superoxide-producing enzymes involved in increased oxidative stress within vascular tissue include the NAD(P)H oxidase, the xanthine oxidase, and mitochondrial superoxide-producing enzymes. Superoxide produced by the NADPH oxidase may react with NO released by endothelial nitric oxide synthase (eNOS), thereby generating peroxynitrite. Peroxynitrite in turn has been shown to uncouple eNOS, thereby switching an antiatherosclerotic NO-producing enzyme to an enzyme that may initiate or even accelerate the atherosclerotic process by producing superoxide. Increased oxidative stress in the vasculature, however, is not restricted to the endothelium and has also been demonstrated to occur within the smooth muscle cell layer in the setting of hypercholesterolemia, diabetes mellitus, hypertension, congestive heart failure, and nitrate tolerance. Increased superoxide production by the endothelial and/or smooth muscle cells has important consequences with respect to signaling by the soluble guanylyl cyclase (sGC) and the cGMP-dependent protein kinase I (cGK-I), the activity and expression of which has been shown to be regulated in a redox-sensitive fashion. The present review summarizes current concepts concerning eNOS uncoupling and also focuses on the consequences for downstream signaling with respect to activity and expression of the sGC and cGK-I in various diseases. PMID- 15879306 TI - Functional properties of mouse connexin30.2 expressed in the conduction system of the heart. AB - Gap junction channels composed of connexin (Cx) 40, Cx43, and Cx45 proteins are known to be necessary for impulse propagation through the heart. Here, we report mouse connexin30.2 (mCx30.2) to be a new cardiac connexin that is expressed mainly in the conduction system of the heart. Antibodies raised to the cytoplasmic loop or the C-terminal regions of mCx30.2 recognized this protein in mouse heart as well as in HeLa cells transfected with wild-type mCx30.2 or mCx30.2 fused with enhanced green fluorescent protein (mCx30.2-EGFP). Immunofluorescence analyses of adult hearts yielded positive signals within the sinoatrial node, atrioventricular node, and A-V bundle of the cardiac conduction system. Dye transfer studies demonstrated that mCx30.2 and mCx30.2-EGFP channels discriminate poorly on the basis of charge, but do not allow permeation of tracers >400 Da. Both mCx30.2 and mCx30.2-EGFP gap junctional channels exhibited weak sensitivity to transjunctional voltage (Vj) and a single channel conductance of approximately 9 pS, which is the lowest among all members of the connexin family measured in HeLa cell transfectants. HeLa mCx30.2-EGFP transfectants when paired with cells expressing Cx40, Cx43, or Cx45 formed functional heterotypic gap junction channels that exhibited low unitary conductances (15 to 18 pS), rectifying open channel I-V relations and asymmetric Vj dependence. The electrical properties of homo- and hetero-typic junctions involving mCx30.2 may contribute to slow propagation velocity in nodal tissues and directional asymmetry of excitation spread in the AV nodal region. PMID- 15879307 TI - p38 mitogen-activated protein kinase activates eNOS in endothelial cells by an estrogen receptor alpha-dependent pathway in response to black tea polyphenols. AB - Black tea has been shown to improve endothelial function in patients with coronary artery disease and recent data indicate the polyphenol fraction of black tea enhances endothelial nitric oxide synthase (eNOS) activity through p38 MAP kinase (p38 MAPK) activation. Because the mechanisms for this phenomenon are not yet clear, we sought to elucidate the signaling events in response to black tea polyphenols. Bovine aortic endothelial cells (BAECs) exposed to black tea polyphenols demonstrated eNOS activation that was inhibited by the estrogen receptor (ER) antagonist ICI 182,780, and siRNA-mediated silencing of ER expression. Consistent with this observation, black tea polyphenols induced time dependent phosphorylation of ERalpha on Ser-118 that was inhibited by ICI 182,780. Phosphorylation of ERalpha on Ser-118 was due to p38 MAP kinase (p38 MAPK) as, it was inhibited by SB203580 and overexpression of dominant-negative p38alpha MAPK. Conversely, constitutively active MKK6 induced p38 MAPK activation that recapitulated the effects of polyphenols by inducing ERalpha phosphorylation and downstream activation of Akt, and eNOS. The key role of ERalpha Ser-118 phosphorylation was confirmed in eNOS-transfected COS-7 cells, as polyphenol induced eNOS activation required cotransfection with ERalpha subject to phosphorylation at Ser-118. This residue appeared critical for functional association of ERalpha with p38 MAPK as ERalpha with Ser-118 mutated to alanine could not form a complex with p38 MAPK. These findings suggest p38 MAP kinase mediated eNOS activation requires ERalpha and these data uncover a new mechanism of ERalpha activation that has broad implications for NO bioactivity and endothelial cell phenotype. PMID- 15879308 TI - BMK1/ERK5 is a novel regulator of angiogenesis by destabilizing hypoxia inducible factor 1alpha. AB - Big MAP kinase 1 (BMK1 or ERK5) is a key mediator of endothelial cell (EC) function as shown by impaired embryonic angiogenesis and vascular collapse in BMK1 knockout mice. Hypoxia inducible factor 1alpha (HIF1alpha), a potent mediator of angiogenesis, is positively regulated by the MAP kinases, ERK1/2. Because BMK1 deficiency is associated with impaired angiogenesis we hypothesized that BMK1 might regulate HIF1alpha. To test this hypothesis, bovine lung microvascular ECs (BLMECs) were transfected with HIF1alpha and BMK1 cDNAs, and stimulated by hypoxia. HIF1alpha activity was measured by a reporter gene assay in which luciferase expression was driven by HIF1alpha activation. Hypoxia (1% O2, 24 hours) stimulated HIF1alpha activity by 5.1+/-0.6 fold. In the presence of dominant negative (DN)-BMK1, which inhibited BMK1 activity, hypoxia induced HIF1alpha activity was enhanced significantly to 6.4+/-0.4 fold. BMK1 activation by constitutively active (CA)-MEK5 inhibited HIF1alpha activity by 46+/-4%, suggesting BMK1 functions as a negative regulator of HIF1alpha activation. Activation of BMK1 reduced HIF1alpha protein levels. Ubiquitination inhibitors (30 micromol/L ALLN, 2 micromol/L lactacystin, or 100 nmol/L MG132) reduced the BMK1-mediated effect on HIF1alpha expression by >80%, suggesting that BMK1 stimulated HIF1alpha proteolysis. The negative effect of BMK1 on HIF1alpha was functionally important because transfection with CA-MEK5 significantly decreased EC migration by 68+/-10%, and inhibited angiogenesis (in vitro Matrigel assay) by 76+/-7%. In summary, BMK1 is a novel negative regulator of HIF1alpha and angiogenesis by increasing HIF1alpha ubiquitination and inhibiting HIF1alpha activity in endothelial cells. PMID- 15879309 TI - Oxygen regulation of tumor perfusion by S-nitrosohemoglobin reveals a pressor activity of nitric oxide. AB - In erythrocytes, S-nitrosohemoglobin (SNO-Hb) arises from S-nitrosylation of oxygenated hemoglobin (Hb). It has been shown that SNO-Hb behaves as a nitric oxide (NO) donor at low oxygen tensions. This property, in combination with oxygen transport capacity, suggests that SNO-Hb may have unique potential to reoxygenate hypoxic tissues. The present study was designed to test the idea that the allosteric properties of SNO-Hb could be manipulated to enhance oxygen delivery in a hypoxic tumor. Using Laser Doppler flowmetry, we showed that SNO-Hb infusion to animals breathing 21% O2 reduced tumor perfusion without affecting blood pressure and heart rate. Raising the pO2 (100% O2) slowed the release of NO bioactivity from SNO-Hb (ie, prolonged the plasma half-life of the SNO in Hb), preserved tumor perfusion, and raised the blood pressure. In contrast, native Hb reduced both tumor perfusion and heart rate independently of the oxygen concentration of the inhaled gas, and did not elicit hypertensive effects. Window chamber (to image tumor arteriolar reactivity in vivo) and hemodynamic measurements indicated that the preservation of tissue perfusion by micromolar concentrations of SNO-Hb is a composite effect created by reduced peripheral vascular resistance and direct inhibition of the baroreceptor reflex, leading to increased blood pressure. Overall, these results indicate that the properties of SNO-Hb are attributable to allosteric control of NO release by oxygen in central as well as peripheral issues. PMID- 15879310 TI - Cell-to-cell connection of endothelial progenitor cells with cardiac myocytes by nanotubes: a novel mechanism for cell fate changes? AB - The regeneration of new myocardium by stem or progenitor cells is an important therapeutic option. Cellular or nuclear fusion is considered as an alternative to cell reprogramming by transdifferentiation. However, the generation of hybrid cells may also be a consequence of a transient transmembrane exchange of proteins and organelles between cells. Therefore, we investigated the formation of intercellular connections, which may allow the transport of macromolecular structures between labeled adult human endothelial progenitor cells (EPC) and GFP expressing neonatal rat cardiomyocytes (CM) in a coculture system. FACS analysis revealed that, 6 days after initiation of coculture, 2.1+/-0.4% of the cells stained positive for GFP and Dil-ac-LDL. 6 hours after initiation of the coculture, ultrafine intercellular structures between Dil-ac-LDL-labeled EPC and GFP-expressing CM were observed. The number of EPC, which established nanotubular connections with CM increased from 0.5+/-0.2% after 6 hours to 2.6+/-0.3% after 24 hours of coculture. The intercellular connections had a diameter from 50 to 800 nm, a length of 5 to 120 mum, and were only transiently established. To determine whether the nanotubular structures allowed the transport of organelles, we labeled CM with a mitochondrial live tracker (MitoTracker). Using time-lapse video microscopy, we observed the transport of stained complexes between CM and EPC resulting in up-take of MitoTracker-positive structures in EPC. Thus, the present study shows a novel type of cell-to-cell communication between progenitor cells and CM in vitro, which may contribute to the acquisition of a cardiomyogenic phenotype independent of permanent cellular or nuclear fusion. PMID- 15879311 TI - Diabetes induces endothelial dysfunction but does not increase neointimal formation in high-fat diet fed C57BL/6J mice. AB - Studies of diabetic vascular disease have traditionally used murine models of type 1 diabetes and genetic models of type 2 diabetes. Because the majority of patients with type 2 diabetes have diet induced obesity, we sought to study the effect of diabetes on arterial disease in a mouse model of diet induced obesity/diabetes. C57Bl/6 mice fed a high-fat diet for 9 weeks developed type 2 diabetes characterized by elevated body weight, hyperglycemia, and hyperinsulinemia. Arteries from diabetic mice exhibited a marked decrease in endothelium-dependent vasodilation, a modest decrease in endothelium independent vasodilation, and an increase in sensitivity to adrenergic vasoconstricting agents. Insulin stimulated protein kinase B (akt) and endothelial nitric oxide synthase (eNOS) phosphorylation were preserved in arteries from diabetic mice; however, eNOS protein dimers were markedly diminished. Arterial nitrotyrosine staining indicated that increased levels of peroxynitrite contributed to eNOS dimer disruption in the diabetic mice. The abnormal vasomotion was not an acute response to the high-fat diet, as short term high-fat diet feeding had no effect on endothelium dependent dilation. A trend toward smaller neointimal lesions was noted in high-fat diet fed mice after femoral artery wire denudation injury. In summary, disrupted eNOS dimer formation rather than impaired insulin mediated eNOS phosphorylation contributed to the endothelial dysfunction in diet induced obese/diabetic mice. The lack of an increase in neointimal formation indicates that additional diabetes associated parameters (such as hyperlipidemia and atherosclerotic vascular disease) may need to be present to increase neointimal formation in this model. PMID- 15879312 TI - Endotoxin-induced myocardial dysfunction: effects of macrophage migration inhibitory factor neutralization. AB - The pathophysiology of sepsis-induced myocardial dysfunction still remains controversial. Macrophage migration inhibitory factor (MIF) has recently been identified as a cardiac-derived myocardial depressant factor in septic shock. Putative mechanisms by which MIF affects cardiac function are unknown. In an investigation of possible mechanisms of action, a rat model of endotoxin toxicity was designed using intraperitoneal (I/P) injection of lipopolysaccharides (LPS) with or without coinfusion of neutralizing anti-MIF or isotypic-matched antibodies. Echocardiographic evaluation revealed that MIF neutralization reversed endotoxin-induced myocardial dysfunction at 24 hours after injection. RNase protection assay (RPA) and Western blot established that MIF neutralization prevented LPS-induced mRNA expression and production of heart-derived inflammatory paracrine and autocrine cytokines such as IL-1s and IL-6. Moreover, MIF immunoneutralization increased heart Bcl-2/Bax protein ratio and suppressed endotoxin-induced release of mitochondrial cytochrome-c, as demonstrated by Western blotting. Inhibition of mitochondrial loss of cytochrome-c decreased in heart caspase-3 activity at 6 and 24 hours after injection. MIF neutralization also restored the LPS-induced deficient nuclear translocation of phospho-Akt and consequently the expression of the heart survival nuclear factor GATA-4. The restoration of the translocation/expression of survival factors by MIF inhibition resulted in lowered endotoxin-induced DNA fragmentation at 24 hours, a hallmark of downstream cardiomyocyte apoptosis. Our data indicate that early inactivation of MIF significantly reverses the imbalance of proapoptotic to prosurvival pathways and reduces acute inflammation of the heart thereby improving myocardial dysfunction induced by endotoxin. PMID- 15879313 TI - Functional characterization of connexin43 mutations found in patients with oculodentodigital dysplasia. AB - Specific mutations in GJA1, the gene encoding the gap junction protein connexin43 (Cx43), cause an autosomal dominant disorder called oculodentodigital dysplasia (ODDD). Here, we characterize the effects of 8 of these mutations on Cx43 function. Immunochemical studies have shown that most of the mutant proteins formed gap junction plaques at the sites of cell-cell apposition. However, 2 of the mutations (a codon duplication in the first extracellular loop, F52dup, and a missense mutation in the second extracellular loop, R202H, produced full-length connexins that failed to properly form gap junction plaques. Cx43 proteins containing ODDD mutations found in the N-terminus (Y17S), first transmembrane domain (G21R, A40V), second transmembrane domain (L90V), and cytoplasmic loop (I130T, K134E) do form gap junction plaques but show compromised channel function. L90V, I130T, and K134E demonstrated a significant decrease in junctional conductance relative to Cx43WT. Mutations Y17S, G21R, and A40V demonstrated a complete lack of functional electrical coupling even in the presence of significant plaque formation between paired cells. Heterologous channels formed by coexpression of Cx43WT and mutation R202H resulted in electrically functional gap junctions that were not permeable to Lucifer yellow. Therefore, the mutations found in ODDD not only cause phenotypic variability, but also result in various functional consequences. Overall, our data show an extensive range of molecular phenotypes, consistent with the pleiotropic nature of the clinical syndrome as a whole. PMID- 15879320 TI - Why human color vision cannot reliably detect cerebrospinal fluid xanthochromia. AB - BACKGROUND: Visual assessment of cerebrospinal fluid (CSF) for xanthochromia (yellow color) is practiced by the majority of laboratories worldwide as a means of diagnosing intracranical bleeds. METHODS: Colorimetric and spectrophotometric analysis of CSF samples for recognizing the presence of bilirubin either in low concentrations or in the presence of hemolysed blood. RESULTS: The experiments provide the physiological and colorimetric basis for abandoning visual assessment of CSF for xanthochromia. CONCLUSIONS: We strongly recommend relying on spectrophotometry as the analytical method of choice. PMID- 15879321 TI - Steps after stroke: capturing ambulatory recovery. AB - BACKGROUND AND PURPOSE: Nearly two thirds of stroke survivors have deficits impairing ambulatory recovery. Conventional mobility outcome measures such as timed walks and functional independence measure (FIM) do not quantify free-living ambulatory behavior. This study compared step activity monitoring (SAM) with established instruments to assess ambulatory recovery across the outpatient subacute stroke rehabilitation phase. METHODS: We measured FIM mobility subscale, SAM-derived daily steps, Stroke Impact Scale (SIS) mobility scores, and timed walks in 11 subjects after discharge from inpatient rehabilitation and again 3 months later. RESULTS: Significant improvement was measured in free-living step activity (mean gain 80%; P=0.001) but not with timed walks (P=0.4), FIM (P=0.08), or SIS mobility scales (P=0.3). CONCLUSIONS: Microprocessor-linked SAM is a sensitive indicator of ambulatory recovery that measures improvements not captured by other conventional outcome instruments. PMID- 15879322 TI - Association of blood pressure indices and stroke mortality in isolated systolic hypertension. AB - BACKGROUND AND PURPOSE: Isolated systolic hypertension (ISH), systolic blood pressure (BP) > or =160 mm Hg and diastolic BP (DBP) <90 mm Hg, is associated with stroke; however, the correlation between specific BP indices and stroke mortality in ISH is not defined. METHODS: In a pooled analysis of 9 epidemiological studies, we examined whether pulse pressure (PP) was more predictive of stroke mortality than systolic BP (SBP), DBP, or mean BP (MAP) in persons with ISH. Subjects (n=682; 29% male; 77% white; mean age 63.6 years) with ISH, free of cardiovascular disease, and not on antihypertensive drug therapy at baseline were followed a mean of 13.0+/-7.3 years, and 54 stroke deaths occurred. The relative importance of each BP index was compared by the decrease in the -2 log likelihood (a measure of model agreement with data) because of the addition of 1 or a combination of BP indices to a Cox regression model. Hazards ratios (HRs) for fatal stroke for a 1-SD in BP index were determined. RESULTS: PP was the best predictor of stroke mortality based on the decrease in the -2 log likelihood (10.65; P=0.001; HR=1.52), followed by SBP (7.19; P=0.007; HR=1.40), DBP (2.76; P=0.10; HR=0.80), or MAP (0.39; P=0.39; HR=1.10). Any combination of BP indices did not exceed a decrease in the -2 log likelihood of 10.72. CONCLUSIONS: These data suggest that in persons with ISH, PP is a better predictor of fatal stroke than SBP, DBP, or MAP. PMID- 15879323 TI - Early prediction of gross hemorrhagic transformation by noncontrast agent MRI cluster analysis after embolic stroke in rat. AB - BACKGROUND AND PURPOSE: Our goal was to develop magnetic resonance indices, without image contrast agent enhancement, that predict hemorrhagic transformation (HT) in a rat model of embolic stroke. METHODS: Male Wistar rats subjected to embolic stroke with (n=12) or without (n=10) the combination treatment with recombinant tissue plasminogen activator and an anti-platelet glycoprotein IIb/IIIa antibody 7E3 F(ab')2 initiated at 4 hours after onset of stroke were investigated using a 7-T MRI system. Radiofrequency saturation T1 (T1sat) maps with magnetization transfer, apparent diffusion coefficient of water (ADCw) maps in 3 directions, and T2 maps were measured at 2, 24, and 48 hours after embolization. MRI data were analyzed individually and using 2D cluster plots. Histological measurements were obtained at 48 hours. RESULTS: Gross hemorrhage was detected at 48 hours in 7 (4 control, 3 treated) of 22 animals. The 2D cluster plot using MRI T1sat and ADCw maps obtained at 2 hours after stroke predicted all gross HT. The location of gross hemorrhage predicted by the 2D cluster plot was within 0.75 mm of the identifying MRI cluster. CONCLUSIONS: The 2D MRI cluster plot analysis using T1sat and ADCw maps acquired at 2 hours after the onset of embolic stroke predicts gross HT. PMID- 15879324 TI - Enriched environment increases neural stem/progenitor cell proliferation and neurogenesis in the subventricular zone of stroke-lesioned adult rats. AB - BACKGROUND AND PURPOSE: The subventricular zone in the adult brain is identified as an endogenous resource of neuronal precursors that can be recruited to adjacent lesioned areas. The hypothesis was tested that postischemic environmental enrichment might enhance subventricular zone cell genesis. METHODS: A cortical infarct was induced in adult spontaneously hypertensive rats by ligating the middle cerebral artery distal to the striatal branches, after which animals were housed in either standard or enriched environment and allowed to survive for 5 weeks. The thymidine analogue bromodeoxyuridine was administered during the first postischemic week. The generation of neural stem/progenitor cells and neuronal precursors in the subventricular zone were studied with cell specific markers such as Ki67 and phosphorylated histone H3 (cell proliferation), Sox-2 (neural stem/progenitor cells), bromodeoxyuridine (slowly cycling, nonmigratory putative neural stem cells), and doublecortin (newborn immature neurons). RESULTS: Proliferating cells in the subventricular zone were identified as chiefly neural progenitors but also putative neural stem cells and neuronal precursors. Five weeks after stroke, proliferation in the subventricular zone was lower in stroke-lesioned rats housed in standard environment compared with nonlesioned rats. Postischemic environmental enrichment normalized cell proliferation levels, increased the numbers of putative neural stem cells as assessed with bromodeoxyuridine, and increased doublecortin-positive neuroblasts, which extended in migratory chains toward the infarct. CONCLUSIONS: Enriched environment increased the neural stem/progenitor cell pool and neurogenesis in the adult subventricular zone 5 weeks after a cortical stroke. This might be of potential importance for tissue regeneration. PMID- 15879325 TI - Stroke in south west Nigeria: a 10-year review. AB - BACKGROUND AND PURPOSE: Stroke is a significant economic, social, and medical problem worldwide. This retrospective follow-up study aimed to review the pattern, types, and case fatality of stroke in Nigeria. METHODS: Records of all stroke patients admitted into Ogun State University Teaching Hospital (OSUTH), Sagamu, from December 1993 to November 2003 were reviewed. Patients were classified into hemorrhage or infarct using the World Health Organization criteria. Information was obtained as to the time of death in those who died and case fatality at 24 hours, 7 days, 30 days, and 6 months recorded. Autopsy records were also reviewed. RESULTS: A total of 708 stroke patients were reviewed and this constituted 2.4% of all emergency admissions. On clinical grounds, 49% of the patients had cerebral infarction (CI) and 45% had intracerebral hemorrhage (ICH), whereas 6% had subarachnoid hemorrhage. Stroke constituted 1.8% of all deaths at the emergency unit and the case fatality was 9% at 24 hours, 28% at 7 days, 40% at 30 days, and 46% at 6 months. CONCLUSIONS: Stroke constitutes a significant cause of mortality and the need for prompt institution of intensive treatment is emphasized. A changing pattern with an increasing frequency of hemorrhagic stroke in our population is suspected. However, because this was a retrospective study based on clinical examination in a highly selected stroke population, neuroimaging confirmation would be needed for any future prospective hospital or population-based studies. PMID- 15879326 TI - Regional impairment of cerebrovascular reactivity and BOLD signal in adults after stroke. AB - BACKGROUND AND PURPOSE: Comparative studies across populations using functional magnetic resonance imaging (fMRI) rely on a similar relationship between blood oxygen level-dependent (BOLD) signal and neural activity. However, in elderly and patients with cerebrovascular disease, impaired cerebrovascular dynamics and neurovascular coupling may explain differences in BOLD contrast across populations and brain regions. The purpose of the study was to determine whether poststroke patients have regional heterogeneities of cerebrovascular reactivity (CVR) and their potential influence on voxel-wise motor-related BOLD signal. METHODS: Using fMRI, 8 fully recovered patients from stroke in the frontal lobe without cortical lesion in the regions of interest located in the primary sensorimotor cortex (SMC), supplementary motor area (SMA), and cerebellum (CRB) were compared with 8 healthy subjects. Motor-related BOLD signal changes (%SC) were evaluated during simple unimanual and bimanual tasks, and CVR was evaluated during hyperventilation (HV). Analyses were performed using Lipsia software in SMC, SMA, and CRB. RESULTS: In controls, amplitudes of BOLD signal were symmetrical in all regions of interest during all motor tasks and HV. In patients, %SC was decreased in SMC and SMA of the lesioned hemisphere despite their apparent anatomical integrity for all tasks. Impaired CVR was a predictor of impaired motor-related BOLD response in the SMC during contralateral movements (beta=-1.87; R=-0.75; P=0.03). CONCLUSIONS: These preliminary findings suggest that CVR heterogeneities may account for task-related BOLD signal changes in patients after stroke. PMID- 15879327 TI - Embolic signals and prediction of ipsilateral stroke or transient ischemic attack in asymptomatic carotid stenosis: a multicenter prospective cohort study. AB - BACKGROUND AND PURPOSE: We tested the hypothesis that transcranial Doppler embolic signal (ES) detection identifies an increased risk of ipsilateral carotid stroke or transient ischemic attack (TIA) in subjects with asymptomatic severe carotid stenosis. METHODS: Subjects with duplex-determined 60% to 99% carotid stenosis, without other apparent cerebroembolic sources, underwent 6-monthly neurological assessment and 60-minute ES monitoring. ES positivity was defined as > or =1 ES detected in > or =1 study, ES negativity as no ES in any study, and consistent ES negativity as no ES in any study where > or =6 studies were performed. Rates of ipsilateral carotid stroke/TIA were calculated using Kaplan Meier analysis and correlated with ES status using odds ratios (ORs) and Cox proportional hazards regression analysis. RESULTS: A total of 202 subjects (138 male; mean age 74 years; mean follow-up 34 months) were recruited. The average annual rate of ipsilateral carotid stroke/TIA was 3.1%. A total of 231 arteries were monitored at least once (mean 4.3 studies/artery). Six of 60 (10.0%) ES positive arteries had an ipsilateral carotid stroke/TIA compared with 12 of 171 (7.0%) ES-negative arteries (OR, 1.47; 95% CI, 0.43, 4.48; P=0.624) and 2 of 41 (4.9%) consistently ES-negative arteries (OR, 2.17; 95% CI, 0.36, 22.90; P=0.59). Differences in survival free of ipsilateral carotid stroke/TIA according to ES status were not statistically significant. CONCLUSIONS: Although there were more ipsilateral carotid cerebrovascular events among ES-positive arteries, this was not statistically significant. Less labor-intensive techniques are required to make further study and clinical application practical. PMID- 15879328 TI - Protein Z gene polymorphisms, protein Z concentrations, and ischemic stroke. AB - BACKGROUND AND PURPOSE: We aimed to determine whether A-13G or G79A polymorphisms of the protein Z gene that have been reported to be an important determinant of blood concentrations of protein Z are associated with risk of ischemic stroke in a broad range of stroke patients and controls. METHODS: We conducted a case control study of 151 hospital cases of first-ever ischemic stroke and 164 randomly selected community controls. Protein Z genotype was determined for the A 13G promoter polymorphism and the G79A intron F polymorphism, and plasma protein Z concentrations were measured during the first 7 days and at 3 to 6 months after the acute stroke event. RESULTS: Geometric mean concentrations of protein Z measured within 7 days of acute stroke were significantly higher in cases compared with controls (1.51 microg/mL versus 1.13 microg/mL; P<0.0001). Protein Z concentrations were highest among subjects with the A-13G AA genotype, intermediate among those with the AG genotype, and lowest among those with the GG genotype (1.39 microg/mL versus 1.05 microg/mL versus 0.76 microg/mL; P<0.0001); and highest among those with the G79A GG genotype, intermediate among those with the GA genotype, and lowest among those with the AA genotype (1.47 microg/mL versus 1.13 microg/mL versus 0.66 microg/mL; P<0.0001). The prevalence of A-13G and G79A genotypes was not significantly different between cases of ischemic stroke and controls. However, compared with the G79A GG genotype (reference), the odds of ischemic stroke was progressively lower for the heterozygote GA (odds ratio [OR], 0.83; 95% CI, 0.52 to 1.33) and the homozygote AA genotype (OR, 0.63; 95% CI, 0.20 to 1.98). A pooled analysis showed that compared with the G79A GG genotype (reference), the odds of ischemic stroke was progressively lower for the heterozygote GA (OR, 0.78; 95% CI, 0.57 to 1.07) and the homozygote AA genotype (OR, 0.31; 95% CI, 0.14 to 0.69). CONCLUSIONS: The consistency of the association between protein Z genotypes, blood concentrations of protein Z, and ischemic stroke, determined using 2 different methods that have different sources of bias strengthens the evidence that increased blood concentrations of protein Z concentrations are associated causally with an increased risk of ischemic stroke. PMID- 15879329 TI - Increased plasma levels of 15-deoxyDelta prostaglandin J2 are associated with good outcome in acute atherothrombotic ischemic stroke. AB - BACKGROUND AND PURPOSE: The 15-deoxyDelta prostaglandin J2 (15-dPGJ2) is an anti inflammatory prostaglandin that has been proposed to be the endogenous ligand of peroxisome proliferator-activated receptor-gamma (PPARgamma), a nuclear receptor that can exert potent anti-inflammatory actions by repressing inflammatory genes when activated. It has been suggested that 15-dPGJ2 could be beneficial in neurological disorders in which inflammation contributes to cell death such as stroke. METHODS: We investigated the relationship between plasma levels of 15 dPGJ2 and early neurological deterioration (END), infarct volume, and neurologic outcome in 552 patients with an acute stroke admitted within 24 hours after symptoms onset. RESULTS: Median [quartiles] plasma 15-dPGJ2 levels on admission were significantly higher in patients than in controls (60.5 [11.2 to 109.4] versus 5.0 [3.8 to 7.2] pg/mL; P<0.0001). Levels of this prostaglandin were also significantly higher in patients with vascular risk factors (history of hypertension or diabetes) and with atherothrombotic infarcts (113.9 [81.6 to 139.7] pg/mL), than in those with lacunar (58.7 [32.7 to 86.2] pg/mL), cardioembolic (12.1 [6.5 to 39.2] pg/mL), or undetermined origin infarcts (11.4 [5.6 to 24.3] pg/mL) (P<0.0001). In the subgroup of patients with atherothrombotic infarcts, the adjusted odds ratio of END and poor outcome for 1 pg/mL increase in 15-dPGJ2 were 0.95 (95% CI, 0.94 to 0.97) and 0.97 (95% CI, 0.96 to 0.98), respectively. In a generalized linear model, by 1 U increase in 15 dPGJ2, there was a reduction of 0.47 mL (95% CI, 0.32 to 0.63) in the mean estimated infarct volume. CONCLUSIONS: Increased plasma 15-dPGJ2 concentration is associated with good early and late neurological outcome and smaller infarct volume. These findings suggest a neuroprotective effect of 15-dPGJ2 in atherothrombotic ischemic stroke. PMID- 15879330 TI - Contribution of atrial fibrillation to incidence and outcome of ischemic stroke: results from a population-based study. AB - BACKGROUND AND PURPOSE: Atrial fibrillation (AF) is a major risk factor for ischemic stroke and its prevalence increases steeply with age. Population-based data on its influence on stroke outcome are scarce. METHODS: We evaluated the prevalence of AF and its influence on prognosis in patients with a first-ever ischemic stroke from a population-based registry. RESULTS: The presence of AF at stroke onset and during the acute phase was confirmed by a standard electrocardiogram in 869 (24.6%) of 3530 patients with ischemic stroke. With respect to patients without the arrhythmia, those with AF were more frequently women, aged 80 years and older, with coronary heart disease and peripheral arterial disease. The presence of AF was associated with high 30-day (32.5%; 95% CI, 29.3 to 35.6) and 1-year case-fatality rates (49.5%; 95% CI, 46.2 to 52.8), with a higher stroke recurrence rate within the first year of follow-up (6.6% versus 4.4%; P=0.046) and with the worst survival after an average follow-up of 45.2 months (P<0.0001). At the multivariate Cox regression analysis, AF was an independent predictor of 30-day and 1-year mortality. Approximately 17% of all deaths were attributable to the presence of AF. CONCLUSIONS: We found a high prevalence of AF in patients with a first-ever ischemic stroke, especially among elderly women. The overall contribution of AF to stroke mortality was relevant, suggesting that together with new strategies to prevent the development of the arrhythmia more appropriate treatments are needed, mostly in elderly women. PMID- 15879331 TI - Vampire bat salivary plasminogen activator (desmoteplase) inhibits tissue-type plasminogen activator-induced potentiation of excitotoxic injury. AB - BACKGROUND AND PURPOSE: In contrast to tissue-type plasminogen activator (tPA), vampire bat (Desmodus rotundus) salivary plasminogen activator (desmoteplase [DSPA]) does not promote excitotoxic injury when injected directly into the brain. We have compared the excitotoxic effects of intravenously delivered tPA and DSPA and determined whether DSPA can antagonize the neurotoxic and calcium enhancing effects of tPA. METHODS: The brain striatal region of wild-type c57 Black 6 mice was stereotaxically injected with N-methyl-d-Aspartate (NMDA); 24 hour later, mice received an intravenous injection of tPA or DSPA (10 mg/kg) and lesion size was assessed after 24 hours. Cell death and calcium mobilization studies were performed using cultures of primary murine cortical neurons. RESULTS: NMDA-mediated injury was increased after intravenous administration of tPA, whereas no additional toxicity was seen after administration of DSPA. Unlike DSPA, tPA enhanced NMDA-induced cell death and the NMDA-mediated increase in intracellular calcium levels in vitro. Moreover, the enhancing effects of tPA were blocked by DSPA. CONCLUSIONS: Intravenous administration of tPA promotes excitotoxic injury, raising the possibility that leakage of tPA from the vasculature into the parenchyma contributes to brain damage. The lack of such toxicity by DSPA further encourages its use as a thrombolytic agent in the treatment of ischemic stroke. PMID- 15879332 TI - Morbidity and Mortality After Stroke, Eprosartan Compared with Nitrendipine for Secondary Prevention: principal results of a prospective randomized controlled study (MOSES). AB - BACKGROUND AND PURPOSE: In hypertensive stroke patients, for the same level of blood pressure control, eprosartan will be more effective than nitrendipine in reducing cerebrovascular and cardiovascular morbidity and mortality. METHODS: A total of 1405 well-defined, high-risk hypertensives with cerebral event during the last 24 months (proven by cerebral computed tomography scan or nuclear magnetic resonance) were randomized to eprosartan or nitrendipine (mean follow-up 2.5 years). Primary end point was the composite of total mortality and all cardiovascular and cerebrovascular events, including all recurrent events. RESULTS: Randomization was successful without significant differences in the baseline characteristics. Blood pressure was reduced to a comparable extent without any significant differences between the 2 groups during the whole study period (150.7/84 mm Hg and 152.0/87.2 mm Hg with eprosartan and nitrendipine therapy to 137.5/80.8 mm Hg and 136.0/80.2 mm Hg, respectively, confirmed by ambulatory blood pressure monitoring). Moreover, already after 3 months, normotensive mean values were achieved, and 75.5% reached values <140/90 mm Hg with the eprosartan regimen and 77.7% with the nitrendipine regimen. During follow-up, in total, 461 primary events occurred: 206 eprosartan and 255 nitrendipine (incidence density ratio [IDR], 0.79; 95% CI, 0.66 to 0.96; P=0.014). Cardiovascular events were: 77 eprosartan and 101 nitrendipine (IDR, 0.75; 95% CI, 0.55 to 1.02; P=0.06); cerebrovascular events: 102 eprosartan and134 nitrendipine (IDR, 0.75; 95% CI, 0.58 to 0.97; P=0.03). CONCLUSIONS: The Morbidity and Mortality After Stroke, Eprosartan Compared With Nitrendipine for Secondary Prevention (MOSES) study was the first to compare an angiotensin II type 1 receptor antagonist with a calcium antagonist in secondary stroke prevention. In these high-risk hypertensive stroke patients, an early normotensive and comparable blood pressure was achieved. The combined primary end point was significantly lower in the eprosartan group. PMID- 15879333 TI - Silent myocardial ischemia in patients with symptomatic intracranial atherosclerosis: associated factors. AB - BACKGROUND AND PURPOSE: Optimization of coronary risk evaluation in stroke patients has been encouraged. The relationship between symptomatic intracranial atherosclerosis and occult coronary artery disease (CAD) has not been evaluated sufficiently. We aimed to investigate the prevalence of silent myocardial ischemia in patients with symptomatic intracranial atherosclerosis and to identify factors associated with its presence. METHODS: From 186 first-ever transient ischemic attack or ischemic stroke patients with intracranial stenoses, 65 fulfilled selection criteria, including angiographic confirmation of a symptomatic atherosclerotic stenosis and absence of known CAD. All patients underwent a maximal-stress myocardial perfusion single-photon emission computed tomography (SPECT). Lipoprotein(a) [Lp(a)], C-reactive protein, and homocysteine (Hcy) levels were determined before SPECT. RESULTS: Stress-rest SPECT detected reversible myocardial perfusion defects in 34 (52%) patients. Vascular risk factors associated with a pathologic SPECT were hypercholesterolemia (P=0.045), presence of >2 risk factors (P=0.004) and high Lp(a) (P=0.023) and Hcy levels (P=0.018). Ninety percent of patients with high Lp(a) and Hcy levels had a positive SPECT. Existence of a stenosed intracranial internal carotid artery (ICA; odds ratio [OR], 7.22, 2.07 to 25.23; P=0.002) and location of the symptomatic stenosis in vertebrobasilar arteries (OR, 4.89, 1.19 to 20.12; P=0.027) were independently associated with silent myocardial ischemia after adjustment by age, sex, and risk factors. CONCLUSIONS: More than 50% of the patients with symptomatic intracranial atherosclerosis and not overt CAD show myocardial perfusion defects on stress-rest SPECT. Stenosed intracranial ICA, symptomatic vertebrobasilar stenosis and presence of high Lp(a) and Hcy levels may characterize the patients at a higher risk for occult CAD. PMID- 15879334 TI - Secondary prevention of stroke is important: but all hypertensive drugs are not created equal? PMID- 15879335 TI - AutoCITE: automated delivery of CI therapy with reduced effort by therapists. AB - BACKGROUND AND PURPOSE: To evaluate the effectiveness of a device that automates Constraint-Induced Movement therapy (CI therapy), termed AutoCITE, when only partially supervised by therapists. METHODS: Twenty-seven participants with chronic stroke trained with AutoCITE for 3 hours per day for 10 consecutive weekdays. Participants were assigned to 1 of 3 groups in a fixed irregular order (ie, in alternating blocks): supervision from a therapist for 100%, 50%, or 25% of training time. RESULTS: The effect sizes of the treatment gains for the 3 groups on the Motor Activity Log (MAL) were very large and for the Wolf Motor Function Test they were large (all P<0.001) but were not significantly different from one another. Gains were comparable to those previously reported for participants who received an equal amount of standard one-on-one CI therapy without the device. At 1-month and long-term follow-up, gains from pretreatment on the MAL were also significant (P<0.001). CONCLUSIONS: These results demonstrate that AutoCITE training with greatly reduced supervision from a therapist is as effective as standard one-on-one CI therapy. PMID- 15879336 TI - Improved quality of stroke care for hospitalized Medicare beneficiaries in Michigan. AB - BACKGROUND AND PURPOSE: We reported previously that acute ischemic stroke patients encountered delays in obtaining neuroimaging and receiving thrombolysis, and that deep venous thrombosis prophylaxis was used only in a minority of eligible patients. We investigated whether these and other measures improved after a quality improvement initiative. METHODS: Medicare fee-for-service ischemic stroke and transient ischemic attack discharges in 136 acute care hospitals in Michigan were identified by International Classification of Diseases, 9th Revision, Clinical Modification codes. Only patients with stroke symptoms persisting for >1 hour and present on arrival were included in the analysis. Seven quality indicators were abstracted from chart review at baseline (discharges between July 1, 1998, and June 30, 1999) and at remeasurement (discharges between January 1, 2001, and June 30, 2001) after an intensive quality improvement initiative throughout Michigan hospitals. Quality indicators were compared at baseline and remeasurement. RESULTS: Indicators of care were determined in 5146 patients at baseline and 4980 patients on remeasurement. Four quality-of-care indicators showed significant improvement on remeasurement: antithrombotic prescribed at discharge (81.9 baseline versus 83.7% remeasurement; P=0.026), avoidance of sublingual nifedipine in patients with acute ischemic stroke (97.1 versus 99.7%; P<0.0001), documentation of a computed tomography (CT)/MRI during hospitalization (98.0 versus 99.1%; P=0.024), and appropriate deep venous thrombosis prophylaxis (13.8 versus 26.9%; P<0.0001). Time to CT/MRI did not significantly change, but time to thrombolysis improved (113 versus 88.5 minutes; P=0.045). CONCLUSIONS: Improvement occurred in several indicators of quality of care in Michigan Medicare beneficiaries presenting with acute stroke symptoms. PMID- 15879337 TI - Increased extracellular K+ concentration reduces the efficacy of N-methyl-D aspartate receptor antagonists to block spreading depression-like depolarizations and spreading ischemia. AB - BACKGROUND AND PURPOSE: Spreading depression (SD)-like depolarizations may augment neuronal damage in neurovascular disorders such as stroke and traumatic brain injury. Spreading ischemia (SI), a particularly malignant variant of SD like depolarization, is characterized by inverse coupling between the spreading depolarization wave and cerebral blood flow. SI has been implicated in particular in the pathophysiology of subarachnoid hemorrhage. Under physiological conditions, SD is blocked by N-methyl-D-aspartate receptor (NMDAR) antagonists. However, because both SD-like depolarizations and SI occur in presence of an increased extracellular K+ concentration ([K+]o), we tested whether this increase in baseline [K+]o would reduce the efficacy of NMDAR antagonists. METHODS: Cranial window preparations, laser Doppler flowmetry, and K+-sensitive/reference microelectrodes were used to record SD, SD-like depolarizations, and SI in rats in vivo; microelectrodes and intrinsic optical signal measurements were used to record SD and SD-like depolarizations in human and rat brain slices. RESULTS: In vivo, the noncompetitive NMDAR antagonist dizocilpine (MK-801) blocked SD propagation under physiological conditions, but did not block SD-like depolarizations or SI under high baseline [K+]o. Similar results were found in human and rat neocortical slices with both MK-801 and the competitive NMDAR antagonist D-2-amino-5-phosphonovaleric acid. CONCLUSIONS: Our data suggest that elevated baseline [K+]o reduces the efficacy of NMDAR antagonists on SD-like depolarizations and SI. In conditions of moderate energy depletion, as in the ischemic penumbra, or after subarachnoid hemorrhage, NMDAR inhibition may not be sufficient to block these depolarizations. PMID- 15879340 TI - The past is the future: innovative designs in acute stroke therapy trials. PMID- 15879341 TI - Evaluation of C-reactive protein measurement for assessing the risk and prognosis in ischemic stroke: a statement for health care professionals from the CRP Pooling Project members. AB - BACKGROUND AND PURPOSE: Several studies have shown, in different populations, that modest elevation of plasma C-reactive protein (CRP) in the range seen in apparently healthy individuals is a strong predictor of future vascular events. Elevated plasma CRP concentrations are also associated with an increased risk of cerebrovascular events and an increased risk of fatal and nonfatal cardiovascular events in ischemic stroke patients. These epidemiological and clinical observations suggest that determination of plasma CRP concentrations could be used as an adjunct for risk assessment in primary and secondary prevention of cerebrovascular disease and be of prognostic value. The aim of this review is to summarize the evidence for CRP as an independent predictor of cerebrovascular events in at-risk individuals and ischemic stroke patients and to consider its usefulness in evaluating prognosis after stroke. SUMMARY OF REVIEW: CRP fulfils most of the requirements of a new risk and prognostic predictor, but several issues await further confirmation and clarification before this marker can be included in the routine evaluation of stroke patients and subjects at risk for cerebrovascular disease. Potentially important associations have been established between elevated plasma CRP concentrations and increased efficacy of established therapies, particularly lipid-lowering therapy with statins. CONCLUSIONS: At present, there is not sufficient evidence to recommend measurement of CRP in the routine evaluation of cerebrovascular disease risk in primary prevention, because there is insufficient evidence as to whether early detection, or intervention based on detection, improves health outcomes, although shared risk of cardiovascular disease indicates this may be of value. In secondary prevention of stroke, elevated CRP adds to existing prognostic markers, but it remains to be established whether specific therapeutic options can be derived from this. PMID- 15879342 TI - Frequency of depression after stroke: a systematic review of observational studies. AB - BACKGROUND AND PURPOSE: Although depression is an important sequelae of stroke, there is uncertainty regarding its frequency and outcome. METHODS: We undertook a systematic review of all published nonexperimental studies (to June 2004) with prospective consecutive patient recruitment and quantification of depressive symptoms/illness after stroke. RESULTS: Data were available from 51 studies (reported in 96 publications) conducted between 1977 and 2002. Although frequencies varied considerably across studies, the pooled estimate was 33% (95% confidence interval, 29% to 36%) of all stroke survivors experiencing depression. Differences in case mix and method of mood assessment could explain some of the variation in estimates across studies. The data also suggest that depression resolves spontaneously within several months of onset in the majority of stroke survivors, with few receiving any specific antidepressant therapy or active management. CONCLUSIONS: Depression is common among stroke patients, with the risks of occurrence being similar for the early, medium, and late stages of stroke recovery. There is a pressing need for further research to improve clinical practice in this area of stroke care. PMID- 15879343 TI - Recurrent stroke risk is higher than cardiac event risk after initial stroke/transient ischemic attack. AB - BACKGROUND AND PURPOSE: Patients with ischemic stroke and transient ischemic attack (TIA) are at risk for recurrent cerebrovascular and cardiac events. Understanding which of these adverse events is more likely to occur next is instructive for preventive therapy planning. METHODS: Subjects (n=1923) were identified from a sample of hospital discharges from administrative claims for the Michigan Medicare population from January 2001 to June 2001 using International Classification of Diseases, 9th Revision codes for ischemic stroke/TIA. Outcomes (cardiac events, myocardial infarction [MI], percutaneous transluminal coronary angioplasty [PTCA], coronary artery bypass grafting [CABG] and ischemic strokes) were identified for 2001 to 2003. Comparison between cardiac and stroke as secondary events were made using cumulative incidence estimates. RESULTS: Over the follow-up period, 172 patients had a cardiac event (62.8% MI, 7.6% CABG, 14.5% PTCA, 9.3% MI and PTCA, and 5.8% MI and CABG) and 239 had a stroke as their first event. Cardiac event at 2 years had occurred in 7.7%, and stroke occurred in 11.8%. CONCLUSIONS: The risk of stroke after initial stroke/TIA is higher than the risk of cardiac events. The propensity after stroke/TIA to have the first recurrent ischemic event in the brain, rather than in the heart, has implications for prophylactic therapy selection. PMID- 15879344 TI - Vascular endothelial growth factor antagonist reduces brain edema formation and venous infarction. AB - BACKGROUND AND PURPOSE: Cerebral venous ischemia often induces severe brain edema. Vascular endothelial growth factor (VEGF), which induces angiogenesis, is also known as vascular permeability (VP) factor. The present study was undertaken to investigate whether the inhibition of VEGF could reduce brain edema formation and cerebral venous infarction (CVI) in a rat 2-vein occlusion (2-VO) model. METHODS: We used 2-VO model in which 2 adjacent cortical veins were photochemically occluded. Male Wistar rats (n=25) were divided into 2 groups: one group was treated with a VEGF antagonist (antagonist group, n=10) and the second group was treated with phosphate-buffered solution (PBS) (PBS group, n=15). VEGF antagonist or PBS was injected intraperitoneally immediately after 2-VO. The developing ischemic infarct was evaluated by magnetic resonance imaging (MRI) and histology 24 hours after occlusion. RESULTS: VEGF expression was observed in the cytoplasm of neurons exclusively in the area of vasogenic edema that was shown as a high-intensity area in the apparent diffusion coefficient of water map. Ischemic volumes calculated from each MR images, which are related to infarction and/or vasogenic edema, respectively, were significantly smaller in the antagonist group as compared with the PBS group (P<0.05) CONCLUSIONS: Our study is the first to provide evidence that the inhibition of VEGF attenuates VP and reduces CVI in the acute stage. Although VEGF is a significant angiogenesis factor, we concluded that the inhibition of VEGF might be a new therapy for both brain edema formation and CVI. PMID- 15879345 TI - Total homocysteine is associated with white matter hyperintensity volume: the Northern Manhattan Study. AB - BACKGROUND: Total homocysteine (tHcy) has been implicated as a risk factor for stroke and dementia, but the mechanism is unclear. White matter hyperintensities may be a risk factor for both, but studies of the relationship between tHcy and quantitative measures of white matter hyperintensity volume (WMHV) are lacking, especially in minority populations. METHODS: A community-based sample of 259 subjects with baseline tHcy levels underwent pixel-based quantitative measurement of WMHV. We examined the relationship between tHcy and WMHV adjusting for age, sociodemographics, vascular risk factors, and B12 deficiency. RESULTS: Higher levels of tHcy were associated with WMHV adjusting for sociodemographics and vascular risk factors. CONCLUSIONS: These cross-sectional data provide evidence that tHcy is a risk factor for white matter damage. PMID- 15879346 TI - HMG-CoA reductase inhibitors improve acute ischemic stroke outcome. AB - BACKGROUND AND PURPOSE: Statins reduce the risk of stroke recurrence, but the benefits of statins in improving outcome of acute stroke patients have not been well explored. METHODS: We assessed potential effects of statins initiated before or within 4 weeks of stroke on 90-day outcome. Favorable outcomes were National Institutes of Health Stroke Scale (NIHSS) score < or =2 at 12 weeks and modified Rankin Scale (mRS) < or =2. RESULTS: Before stroke, 129 patients were receiving statins, 123 initiated statins within 4 weeks, and 600 patients were not on statins. Multivariate logistic regression analysis demonstrated that poststroke statins were associated with a significant probability of a favorable outcome at 12 weeks [NIHSS (P=0.002; OR, 1.92; CI, 1.27 to 2.91) and mRS (P=0.033; OR, 1.57; CI, 1.04 to 2.38)], whereas prestroke statins demonstrated a trend toward significance. CONCLUSIONS: These preliminary results suggest that statin use may improve outcome of acute ischemic stroke. PMID- 15879347 TI - Early carotid atherosclerosis in subjects with periodontal diseases. AB - BACKGROUND AND PURPOSE: There is growing experimental evidence implicating chronic inflammation/infection as an atherosclerotic risk factor. In this study, the involvement of periodontal disease in the development of early atherosclerotic vascular lesions has been evaluated. METHODS: In randomly chosen 82 patients with periodontal disease and 31 periodontally healthy individuals subjected to a clinical oral examination in 1985, atherosclerotic risk factor analysis and carotid ultrasonography was performed during reexamination 16 years later. Common carotid artery intima-media thickness (IMT) and lumen diameter were measured and intima-media area (cIMA) was calculated. The relationship between IMT and cIMA as dependent variables and periodontal disease, age, gender, body mass index, heredity for atherosclerosis, diabetes mellitus, hypertension, plasma cholesterol, smoking, and education as independent variables was evaluated in a multiple logistic regression model. RESULTS: The mean values of IMT and cIMA were significantly higher in patients with periodontal disease than in controls, both at the right (P<0.01 and P<0.001, respectively) and left side (P<0.001 for both variables). When the means of the bilateral measurements of these 2 ultrasonographic variables were tested, multiple logistic regression analysis identified periodontal disease as a principal independent predictor of the common carotid artery cIMA (odds ratio [OR], 5.20; P=0.003) and IMT (OR, 4.64; P=0.004). CONCLUSIONS: The present results indicate that periodontal disease is associated with the development of early atherosclerotic carotid lesions. PMID- 15879348 TI - Production of viral vectors using recombinase-mediated cassette exchange. AB - DNA viruses are often used as vectors for foreign gene expression, but large DNA region from cloned or authentic viral genomes must usually be handled to generate viral vectors. Here, we present a unique system for generating adenoviral vectors by directly substituting a gene of interest in a small transfected plasmid with a replaced gene in a replicating viral genome in Cre-expressing 293 cells using the recombinase-mediated cassette exchange (RMCE) reaction. In combination with a positive selection of the viral cis-acting packaging signal connected with the gene of interest, the purpose vector was enriched to 97.5 and 99.8% after three and four cycles of infection, respectively. Our results also showed that the mutant loxP V (previously called loxP 2272), a variant target of Cre used in the RMCE reaction, was useful as a non-compatible mutant to wild-type loxP. This method could be useful for generating not only a large number of adenovirus vectors simultaneously, but also other DNA virus vectors including helper dependent adenovirus vector. PMID- 15879349 TI - Molecular beacons with a homo-DNA stem: improving target selectivity. AB - Molecular beacons (MBs) are stem-loop DNA probes used for identifying and reporting the presence and localization of nucleic acid targets in vitro and in vivo via target-dependent dequenching of fluorescence. A drawback of conventional MB design is present in the stem sequence that is necessary to keep the MBs in a closed conformation in the absence of a target, but that can participate in target binding in the open (target-on) conformation, giving rise to the possibility of false-positive results. In order to circumvent these problems, we designed MBs in which the stem was replaced by an orthogonal DNA analog that does not cross-pair with natural nucleic acids. Homo-DNA seemed to be specially suited, as it forms stable adenine-adenine base pairs of the reversed Hoogsteen type, potentially reducing the number of necessary building blocks for stem design to one. We found that MBs in which the stem part was replaced by homo adenylate residues can easily be synthesized using conventional automated DNA synthesis. As conventional MBs, such hybrid MBs show cooperative hairpin to coil transitions in the absence of a DNA target, indicating stable homo-DNA base pair formation in the closed conformation. Furthermore, our results show that the homo adenylate stem is excluded from DNA target binding, which leads to a significant increase in target binding selectivity. PMID- 15879350 TI - In vitro selection to identify determinants in tRNA for Bacillus subtilis tyrS T box antiterminator mRNA binding. AB - The T box transcription antitermination regulatory system, found in Gram-positive bacteria, is dependent on a complex set of interactions between uncharged tRNA and the 5'-untranslated mRNA leader region of the regulated gene. One of these interactions involves the base pairing of the acceptor end of cognate tRNA with four bases in a 7 nt bulge of the antiterminator RNA. In vitro selection of randomized tRNA binding to Bacillus subtilis tyrS antiterminator model RNAs was used to determine what, if any, sequence trends there are for binding beyond the known base pair complementarity. The model antiterminator RNAs were selected for the wild-type tertiary fold of tRNA. While there were no obvious sequence correlations between the selected tRNAs, there were correlations between certain tertiary structural elements and binding efficiency to different antiterminator model RNAs. In addition, one antiterminator model selected primarily for a kissing tRNA T loop-antiterminator bulge interaction, while another antiterminator model resulted in no such selection. The selection results indicate that, at the level of tertiary structure, there are ideal matches between tRNAs and antiterminator model RNAs consistent with in vivo observations and that additional recognition features, beyond base pair complementarity, may play a role in the formation of the complex. PMID- 15879351 TI - Exploration of factors driving incorporation of unnatural dNTPS into DNA by Klenow fragment (DNA polymerase I) and DNA polymerase alpha. AB - In order to further understand how DNA polymerases discriminate against incorrect dNTPs, we synthesized two sets of dNTP analogues and tested them as substrates for DNA polymerase alpha (pol alpha) and Klenow fragment (exo-) of DNA polymerase I (Escherichia coli). One set of analogues was designed to test the importance of the electronic nature of the base. The bases consisted of a benzimidazole ring with one or two exocyclic substituent(s) that are either electron-donating (methyl and methoxy) or electron-withdrawing (trifluoromethyl and dinitro). Both pol alpha and Klenow fragment exhibit a remarkable inability to discriminate against these analogues as compared to their ability to discriminate against incorrect natural dNTPs. Neither polymerase shows any distinct electronic or steric preferences for analogue incorporation. The other set of analogues, designed to examine the importance of hydrophobicity in dNTP incorporation, consists of a set of four regioisomers of trifluoromethyl benzimidazole. Whereas pol alpha and Klenow fragment exhibited minimal discrimination against the 5- and 6-regioisomers, they discriminated much more effectively against the 4- and 7 regioisomers. Since all four of these analogues will have similar hydrophobicity and stacking ability, these data indicate that hydrophobicity and stacking ability alone cannot account for the inability of pol alpha and Klenow fragment to discriminate against unnatural bases. After incorporation, however, both sets of analogues were not efficiently elongated. These results suggest that factors other than hydrophobicity, sterics and electronics govern the incorporation of dNTPs into DNA by pol alpha and Klenow fragment. PMID- 15879352 TI - Hypopituitarism after traumatic brain injury. AB - Traumatic brain injury (TBI) is one of the main causes of death and disability in young adults, with consequences ranging from physical disabilities to long-term cognitive, behavioural, psychological and social defects. Post-traumatic hypopituitarism (PTHP) was recognized more than 80 years ago, but it was thought to be a rare occurrence. Recently, clinical evidence has demonstrated that TBI may frequently cause hypothalamic-pituitary dysfunction, probably contributing to a delayed or hampered recovery from TBI. Changes in pituitary hormone secretion may be observed during the acute phase post-TBI, representing part of the acute adaptive response to the injury. Moreover, diminished pituitary hormone secretion, caused by damage to the pituitary and/or hypothalamus, may occur at any time after TBI. PTHP is observed in about 40% of patients with a history of TBI, presenting as an isolated deficiency in most cases, and more rarely as complete pituitary failure. The most common alterations appear to be gonadotropin and somatotropin deficiency, followed by corticotropin and thyrotropin deficiency. Hyper- or hypoprolactinemia may also be present. Diabetes insipidus may be frequent in the early, acute phase post-TBI, but it is rarely permanent. Severity of TBI seems to be an important risk factor for developing PTHP; however, PTHP can also manifest after mild TBI. Accurate evaluation and long-term follow-up of all TBI patients are necessary in order to detect the occurrence of PTHP, regardless of clinical evidence for pituitary dysfunction. In order to improve outcome and quality of life of TBI patients, an adequate replacement therapy is of paramount importance. PMID- 15879353 TI - Debulking of pituitary adenomas improves hormonal control of acromegaly by somatostatin analogues. PMID- 15879354 TI - Effect of long-term continuous methimazole treatment of hyperthyroidism: comparison with radioiodine. AB - OBJECTIVE: To investigate the long-term effects of continuous methimazole (MMI) therapy. DESIGN AND METHODS: Five hundred and four patients over 40 years of age with diffuse toxic goiter were treated with MMI for 18 months. Within one year after discontinuation of MMI, hyperthyroidism recurred in 104 patients. They were randomized into 2 groups for continuous antithyroid and radioiodine treatment. Numbers of occurrences of thyroid dysfunction and total costs of management were assessed during 10 years of follow-up. At the end of the study, 26 patients were still on continuous MMI (group 1), and of 41 radioiodine-treated patients (group 2), 16 were euthyroid and 25 became hypothyroid. Serum thyroid and lipid profiles, bone mineral density, and echocardiography data were obtained. RESULTS: There was no significant difference in age, sex, duration of symptoms and thyroid function between the two groups. No serious complications occurred in any of the patients. The cost of treatment was lower in group 1 than in group 2. At the end of 10 years, goiter rate was greater and antithyroperoxidase antibody concentration was higher in group 1 than in group 2. Serum cholesterol and low density lipoprotein-cholesterol concentrations were increased in group 2 as compared with group 1; relative risks were 1.8 (1.12-2.95, P<0.02) and 1.6 (1.09 2.34, P<0.02) respectively. Bone mineral density and echocardiographic measurements were not different between the two groups. CONCLUSION: Long-term continuous treatment of hyperthyroidism with MMI is safe. The complications and the expense of the treatment do not exceed those of radioactive iodine therapy. PMID- 15879355 TI - Defect of a subpopulation of natural killer immune cells in Graves' disease and Hashimoto's thyroiditis: normalizing effect of dehydroepiandrosterone sulfate. AB - BACKGROUND: The study of the natural killer (NK) immune compartment could provide important findings to help in the understanding of some of the pathogenetic mechanisms related to autoimmune thyroid diseases (Graves' disease (GD) and Hashimoto's thyroiditis (HT)). Within this context, it was suggested that alterations in NK cell cytotoxicity (NKCC) and NK production of cytokines might occur in subjects with GD and HT, whereas the normalization of NK functions could potentially contribute to the prevention of the onset or the progression of both diseases. OBJECTIVE: Due to the hypothesis of alterations in NK in autoimmune thyroid diseases, we were interested to evaluate NKCC in GD and HT patients and to modulate NK function and secretory activity with cytokines and dehydroepiandrosterone sulfate (DHEAS) in an attempt to normalize NK cell defect. DESIGN: We studied 13 patients with recent onset Graves' disease, 11 patients with Hashimoto's thyroiditis at first diagnosis and 15 age-matched healthy subjects. METHODS: NK cells were concentrated at a density of 7.75x10(6) cells/ml by negative immunomagnetic cell separation and validated by FACScan as CD16+/CD56+cells. NK cells were incubated with interleukin-2 (IL-2) and interferon-beta (IFN-beta) and co-incubated with DHEAS at different molar concentrations for measuring NKCC and the secretory pattern of tumor necrosis factor-alpha (TNF-alpha) from NK cells. RESULTS: Lower spontaneous, IL-2- and IFN beta-modulated NKCC was demonstrated in GD and HT patients compared with healthy subjects (P<0.001). A decrease in spontaneous and IL-2-modulated TNF-alpha release from NK cells was also found in both groups of patients (P<0.001). The co incubation of NK cells with IL-2/IFN-beta+DHEAS at different molar concentrations (from 10(-8) to 10(-5) M/ml/NK cells) promptly normalized NKCC and TNF-alpha secretion in GD and HT patients. CONCLUSIONS: A functional defect of a subpopulation of NK immune cells, involving both NKCC and the secretory activity, was demonstrated in newly-diagnosed GD and HT patients. This defect can be reversed by a dose-dependent treatment with DHEAS. The impairment of NK cell activity in autoimmune thyroid diseases could potentially determine a critical expansion of T/B-cell immune compartments leading to the generation of autoantibodies and to the pathogenesis of thyroid autoimmunity. PMID- 15879356 TI - Health status in patients with sub-clinical hypothyroidism. AB - OBJECTIVE: Sub-clinical hypothyroidism (SCH) is a common disorder. People with this condition may have symptoms which could affect their perception of health. Therefore, the perceived health status of people with SCH was assessed and compared with population-matched norms. DESIGN: A prospective cross-sectional survey. METHODS: Seventy-one adults with SCH, age range 18-64 years were studied. Perceived health status was measured by the Short Form-36 (SF-36) version 2 questionaire, which has been validated in a UK population setting. The SF-36 has eight scales measuring physical functioning, role physical, bodily pain, general health, vitality, social functioning, role emotional and mental health. Their SF 36 scores were compared with UK normative data after matching for age and sex and are reported as z-scores. RESULTS: Scores of all eight SF-36 scales were significantly lower in people with SCH compared with the normative population. A negative score (compared with zero of the normative population) indicates worse health status. The most significantly impaired aspects of health status were vitality and role limitations due to physical problems (role physical scale) with z-scores (95% confidence intervals) of -1.01 (-0.74 to -1.29) and -0.73 (-0.43 to -1.04) respectively. Thyroid autoimmunity did not influence the results. CONCLUSION: Perceived health status is significantly impaired in people with SCH when compared with UK normative population scores. This needs to be taken into consideration by clinicians when managing patients with this disease. PMID- 15879357 TI - Discontinuation of estrogen replacement therapy in GH-treated hypopituitary women alters androgen status and IGF-I. AB - OBJECTIVE AND DESIGN: Compared with their male counterparts, healthy females secrete more growth hormone (GH) and those with GH-deficiency have lower insulin like growth factor I (IGF-I) levels and are less responsive to GH substitution. To test whether this gender difference is related to sex hormones we measured androgen status and IGF-I related parameters in 38 hypopituitary women (mean (range) age 41.5 (20-58) years) during continued GH substitution as compared with a control group of 38 healthy women matched for age and menopausal status. Twenty six patients were studied twice: with estrogen replacement and after 28 days of estrogen discontinuation in a randomised design. RESULTS: The patients were androgen deficient compared with controls (median, range), dehydroepiandrosterone sulphate (DHEAS): 185 (99-7800) nmol/l vs 4400 (820-13,000) nmol/l, P=or<0.001; androstenedione: 0.5 (0.1-7.1) nmol/l vs 4.3 (1.6-8.8) nmol/l, P=or<0.001; dihydrotestosterone (DHT): 0.13 (0.09-0.54) nmol/l vs 0.55 (0.09-0.89) nmol/l, P=or<0.001; testosterone: 0.28 (0.09-1.56) nmol/l vs 1.1 (0.71-2.24) nmol/l, (P=or<0.001); free testosterone: 0.004 (0.001-0.030) nmol/l vs 0.016 (0.001 0.030) nmol/l, P=or<0.001. The circulating levels of IGF-I, IGF-II, IGF-binding protein 1 (IGFBP-1), and IGFBP-3 did not differ between patients and controls. The subgroup of patients receiving hydrocortisone (HC) replacement (n=24) had significantly lower levels of androgens (suppressed by 80-100%) as well as IGF-I and IGFBP-3 as compared with the patients not receiving HC. IGF-I was correlated to free testosterone in patients (r=0.57, P=0.0005) as well as controls (r=0.43, P=0.008), and free testosterone was a significant positive predictor of IGF-I. Estrogen discontinuation induced an increase in IGF-I (167+/-15 vs 206+/-14 microg/l, P=0.005 and IGFBP-3 (3887+/-139 vs 4309+/-138 microg/l, P=0.0005). Estrogen discontinuation was associated with a significant increase in median (range) free testosterone (0.004 (0-0.02) vs 0.0065 (0-0.03) nmol/l, P=0.001) and a significant decrease in median (range) sex-hormone binding globulin (SHBG; 93 (11-278) vs 55.5 (20-142) nmol/l, P=0.001). DeltaIGF-I correlated with DeltaSHBG (r=-0.45 P=0.033) and DeltaIGFBP-3 (r=0.67 P=or<0.001). In a regression model DeltaE2, Deltatestosterone, DeltaSHBG and DeltaIGFBP-3 explained 93% of the variation in DeltaIGF-I. CONCLUSIONS: Androgen levels are low in hypopituitary women and free testosterone correlates with IGF-I. Discontinuation of estrogen replacement in these patients induces elevations in IGF-I as well as free testosterone, and DeltaIGF-I correlated positively with Deltafree testosterone. These effects may contribute to the gender differences observed in the GH-IGF axis in healthy adults as well as in the responsiveness of hypopituitary patients to GH substitution. PMID- 15879358 TI - Seasonality of growth response to GH therapy in prepubertal children with idiopathic growth hormone deficiency. AB - OBJECTIVE: Longitudinal growth of children exhibits seasonal variation. In both healthy children and in children with growth hormone (GH) deficiency (GHD) receiving GH therapy, growth rate is maximal during spring and summer. In the present study, we analyzed the growth response to GH therapy in children with GHD as a function of the season when therapy was started. SUBJECTS AND METHODS: Anthropometric measurements and biochemical analyses of GH secretion status and bone formation were longitudinally assessed in a cohort of 52 prepubertal children with GHD (14 girls, mean age 7.6 years) who were treated with a fixed dose of GH (0.025 mg/kg/day). RESULTS: Auxological assessments over the 2-year observation period revealed a significantly better growth response to GH therapy in children who started therapy between the spring and summer (group 1) compared with children who started in the autumn or winter (group 2). The difference was largest in the initial 3-month treatment period (35%; P<0.01). The initial better gain in height of group 1 was sustained during the study period. Baseline peak GH levels during stimulation tests and insuin-like growth factor-I levels did not differ between the two groups. However, group 1 had significantly higher bone resorption and formation markers, either at the start or shortly after initiation of GH treatment. This suggests that children with GHD have higher bone turnover during spring and early summer, irrespective of GH therapy. CONCLUSIONS: In summary, this study suggests that the season of GH initiation is a determinant of the initial growth response to GH replacement in prepubertal children with GHD. PMID- 15879359 TI - Evaluation of adrenal function in patients with growth hormone deficiency and hypothalamic-pituitary disorders: comparison between insulin-induced hypoglycemia, low-dose ACTH, standard ACTH and CRH stimulation tests. AB - OBJECTIVES: Patients with organic growth hormone deficiency (GHD) or with structural hypothalamic-pituitary abnormalities may have additional anterior pituitary hormone deficits, and are at risk of developing complete or partial corticotropin (ACTH) deficiency. Evaluation of the integrity of the hypothalamic pituitary-adrenal axis (HPA) is essential in these patients because, although clinically asymptomatic, their HPA cannot appropriately react to stressful stimuli with potentially life-threatening consequences. DESIGN AND METHODS: In this study we evaluated the integrity of the HPA in 24 patients (age 4.2-31 years at the time of the study) with an established diagnosis of GHD and compared the reliability of the insulin tolerance test (ITT), short synacthen test (SST), low dose SST (LDSST), and corticotropin releasing hormone (CRH) test in the diagnosis of adrenal insufficiency. RESULTS: At a cortisol cut-off for a normal response of 550 nmol/l (20 microg/dl), the response to ITT was subnormal in 11 subjects, 6 with congenital and 5 with acquired GHD. Four patients had overt adrenal insufficiency, with morning cortisol concentrations ranging between 66.2-135.2 nmol/l (2.4-4.9 microg/dl) and typical clinical symptoms and laboratory findings. In all these patients, a subnormal cortisol response to ITT was confirmed by LDSST and by CRH tests. SST failed to identify one of the patients as adrenal insufficient. In the seven asymptomatic patients with a subnormal cortisol response to ITT, the diagnosis of adrenal insufficiency was confirmed in one by LDSST, in none by SST, and in five by CRH tests. The five patients with a normal cortisol response to ITT exhibited a normal response also after LDSST and SST. Only two of them had a normal response after a CRH test. In the seven patients with asymptomatic adrenal insufficiency mean morning cortisol concentration was significantly higher than in the patients with overt adrenal insufficiency. ITT was contraindicated in eight patients, and none of them had clinical symptoms of overt adrenal insufficiency. One of these patients had a subnormal cortisol response to LDSST, SST, and CRH, and three exhibited a subnormal response to CRH but normal responses to LDSST and to SST. CONCLUSION: We conclude that none of these tests can be considered completely reliable for establishing or excluding the presence of secondary or tertiary adrenal insufficiency. Consequently, clinical judgment remains one of the most important issues for deciding which patients need assessment or re-assessment of adrenal function. PMID- 15879360 TI - Inhibitory effect of alcohol on ghrelin secretion in normal man. AB - BACKGROUND: Human appetite is stimulated by alcohol but the underlying mechanism is unknown. It is possible that hunger-stimulating hormones are mediators of this effect of alcohol. Ghrelin stimulates hunger, but how alcohol affects human ghrelin secretion has never been studied before. OBJECTIVE: To investigate whether alcohol ingestion exerts an acute influence on serum ghrelin concentrations in healthy subjects. SUBJECTS AND DESIGN: Eight healthy non-obese subjects participated in the study. All were investigated on two occasions (experiments A and B). Alcohol (0.55 g ethanol/kg body weight) was ingested in experiment A, and drinking-water in experiment B. Venous blood was collected before, and 30 and 60 min after consumption of the drinks. Serum concentrations of ghrelin, cortisol and ethanol were determined and neuropeptide Y (NPY) concentrations were determined in plasma. RESULTS: Alcohol lowered the ghrelin level by 13.9+/-5.0% at 30 min and by 17.5+/-2.6% at 60 min, in contrast to drinking-water which was without significant effect. Serum levels of cortisol and insulin were similar after alcohol and water as was plasma NPY. CONCLUSION: Alcohol has an acute inhibitory influence on human ghrelin secretion but no measurable effect on the secretion of NPY and cortisol. Hence, none of these hormones mediate the orexigenic effect of the drug. PMID- 15879361 TI - Metformin administration improves endothelial function in women with polycystic ovary syndrome. AB - OBJECTIVE: The aim of this study was to investigate the endothelial status in young women with polycystic ovary syndrome (PCOS), using a simple and easily reproducible hemodynamic method combined with a biological marker and to evaluate the effect of metformin treatment on these parameters. DESIGN: Descriptive clinical trial. METHODS: Forty young women, 20 with PCOS and 20 normal women of similar age and body mass index were studied. Metformin (1700 mg daily) was administered for 6 months to the PCOS group. The endothelium status and the metabolic and hormonal profile were studied in both groups, as well as after metformin, by flow-mediated dilatation (FMD) on the brachial artery and by measurements of plasma endothelin-1 (ET-1) levels. RESULTS: FMD was impaired in the PCOS group when compared with controls (3.24+/-0.71% vs 8.81+/-1.07% respectively, P<0.0001), but this difference normalized after metformin treatment (PCOS(post-metformin) vs controls: 8.17+/-1.26 vs 8.81+/-1.07%, P = 0.70) since the values significantly improved after metformin treatment (PCOS(pre-metformin) vs PCOS(post-metformin): 3.24+/-0.71 vs 8.17+/-1.26%, P=0.003). ET-1 levels were significantly higher in the PCOS women compared with the control group (7.23+/ 0.50 vs 4.99+/-0.69 fmol/l, P=0.01), they improved significantly after metformin treatment (PCOS(pre-metformin) vs PCOS(post-metformin): 7.23+/-0.50 vs 3.57+/ 0.60 fmol/l, P<0.0001) and their difference compared with the control group was reversed (PCOS(post-metformin) vs controls: 3.57+/-0.60 vs 4.99+/-0.69 fmol/l, P=0.13). Metformin administration improved hyperandrogenemia. However, in this study, mathematical methods used to assess insulin resistance failed to show any detected alteration after treatment with metformin. CONCLUSIONS: PCOS women were found to exhibit endothelial dysfunction compared with controls, which was reversed 6 months after metformin administration. PMID- 15879362 TI - Octreotide in insulinoma patients: efficacy on hypoglycemia, relationships with Octreoscan scintigraphy and immunostaining with anti-sst2A and anti-sst5 antibodies. AB - OBJECTIVE: We studied the efficacy of octreotide treatment on hypoglycaemia in patients with insulinoma and its relationships with Octreoscan scintigraphy and the presence of tumoral somatostatin receptors sst2A and sst5. DESIGN AND METHODS: 17 patients with insulinoma were evaluated using (i) evaluation of blood glucose, insulin and C-peptide during a short 100 mug octreotide test in fasting patients and/or treatment over 8 days-8 months with octreotide, (ii) Octreoscan scintigraphy and (iii) immunostaining of the tumor with anti-sst2A and anti-sst5. RESULTS: Octreotide was effective on hypoglycaemia in 10/17 patients. Octreoscan scintigraphy detected 4/17 insulinomas. sst2A receptor was detected in 7/17 insulinomas and sst5 in 15/17 insulinomas. Octreotide was effective on hypoglycaemia in those seven patients with sst2A receptor-expressing insulinoma, and in three patients with undetectable sst2A receptor and detectable sst5; it was ineffective in six patients whose tumor expressed the sst5 receptor with undetectable sst2A and in one patient with undetectable sst2A and sst5 receptor. CONCLUSIONS: Octreotide is an effective treatment of hypoglycaemia in more than 50% of patients with insulinoma. Detection of responsive patients was better based on a positive short test with subcutaneous octreotide than on the results of Octreoscan scintigraphy. Positive anti-sst2 receptor immunostaining is associated with efficacy of octreotide treatment, but does not account for all cases of responsiveness to octreotide. Expression of sst5 receptor does not appear to explain per se the efficacy of octreotide on sst2A-negative insulinomas. PMID- 15879363 TI - Clinical and molecular evidence for DAX-1 inhibition of steroidogenic factor-1 dependent ACTH receptor gene expression. AB - BACKGROUND: The ACTH receptor (ACTH-R) is a member of the seven transmembrane domain receptor super-family. In non-functional adrenal adenomas and adrenocortical carcinomas, ACTH-R expression is low. However, no inhibitory factor for ACTH-R expression has been defined to date. DAX-1 (dosage-sensitive sex reversal, adrenal hypoplasia congenita, critical region on the X chromosome, gene-1) is a general repressor of steroid production, inhibiting steroidogenic factor-1 (SF-1)-dependent expression of multiple steroidogenic enzymes. The aim of this study was to investigate whether ACTH-R gene transcription is affected by DAX-1 and whether this mechanism is involved in down-regulation of ACTH-R expression in adrenocortical tumors. METHODS: We screened 22 adrenocortical tumors for ACTH-R and DAX-1 mRNA expression by Northern blot. For in vitro analyses we co-transfected mouse Y1 adrenocortical carcinoma cells with the luciferase reporter gene vector pGL3 containing full-length constructs of human (h) or mouse (m) ACTH-R promoter together with a DAX-1 expression plasmid. These experiments were also performed using ACTH-R promoter 5'-deletion constructs and constructs mutated at the SF-1-binding sites. RESULTS: We found a negative correlation between DAX-1 and ACTH-R mRNA expression (R=-0.47, P<0.02). Accordingly, in vitro expression of DAX-1 significantly reduced hACTH-R and mACTH R promoter activity by 89 and 55% respectively. DAX-1 inhibition was also present in the shortest construct of a series of 5'-deletion constructs of the human promoter extending from -64 to +40 bp relative to the transcription start site. Mutation of the SF-1-binding sites within the hACTH-R promoter resulted in reduced or abolished DAX-1 inhibition, arguing for a mechanism that involves SF-1 for DAX-1 inhibition. CONCLUSIONS: These data support the concept that DAX-1 is a major repressor of ACTH-R gene expression in vitro and in vivo. PMID- 15879364 TI - Evidence for protein and mRNA TSHr expression in fibroblasts from patients with thyroid-associated ophthalmopathy (TAO) after adipocytic differentiation. AB - OBJECTIVE: Thyroid-associated ophthalmopathy (TAO) is a chronic autoimmune disorder characterized by an increased volume of adipose/connective tissue in the human orbit. DESIGN: The aim of this study was to investigate the thyrotropin receptor (TSHr) expression in orbital fibroblasts from TAO patients undergoing adipocytic differentiation. METHODS: Retro-ocular tissue and skin were obtained from five patients undergoing orbital decompression surgery for TAO and placed in culture. Proliferating fibroblasts were subjected to adipocytic differentiation for 10 days. Total RNA was isolated from fibroblasts and was reverse transcribed. TSHr mRNA levels were determined by real-time PCR. cAMP was determined by radioimmunoassay (RIA) after fibroblast incubation with the substances to test. RESULTS: Orbital differentiated fibroblasts became rounded and acquired lipid droplets. The amount of TSHr mRNA in these fibroblasts was higher than fibroblasts not subjected to adipocytic differentiation. Immunocytochemical analysis showed TSHr protein in differentiated orbital fibroblasts. Differentiated orbital fibroblasts stimulated with bovine (b) TSH showed a cAMP production greater than that in paired undifferentiated cultures. A specific thyroid-inhibiting antibody (TBAb) inhibited cAMP production after bTSH challenge, and a thyroid-stimulating antibody (TSAb) stimulated cAMP production in differentiated fibroblasts. CONCLUSIONS: We suggest that orbital fibroblasts subjected to adipocytic differentiation increase TSHr expression that responds specifically to bTSH and TSAb stimulation, and to TBAb inhibition. PMID- 15879365 TI - Evaluation of insulin-like growth factor II, cyclooxygenase-2, ets-1 and thyroid specific thyroglobulin mRNA expression in benign and malignant thyroid tumours. AB - OBJECTIVE: We evaluated three markers (insulin-like growth factor II (IGF-II), cyclooxygenase-2 (COX-2) and ets-1) of thyroid growth stimulation and cell transformation together with a thyroid-specific marker (thyroglobulin (Tg)) for their potential to differentiate benign and malignant follicular thyroid neoplasia (FN). DESIGN AND METHODS: mRNA expression levels were determined by real-time PCR in 100 snap-frozen thyroid samples: 36 benign thyroid nodules with different histology and function (19 cold (CTN) and 17 toxic thyroid nodules (TTN)), 36 corresponding normal thyroid tissues of the same patients, eight Graves' disease (GD) thyroids, 10 follicular thyroid carcinomas (FTC) and 10 papillary thyroid carcinomas (PTC). RESULTS: Mean IGF-II and COX-2 levels were not significantly altered between benign and malignant thyroid nodules (IGF-II) or nodular (FTC, TTN, CTN) and normal thyroid tissues (COX-2). In contrast, eight to tenfold upregulation of ets-1 was observed in PTC and three- to fourfold upregulation of ets-1 was observed in FTC (and GD) compared with benign thyroid nodules and normal thyroid tissues. In addition, thyroglobulin mRNA expression was markedly downregulated (50- to 100-fold) in FTC, PTC and GD samples compared with benign nodular and normal thyroid tissues. Hence an ets-1/Tg ratio >20 distinguished differentiated thyroid cancer from benign nodular or normal thyroid tissue. We then studied ets1- and Tg mRNA expression levels in fine needle aspiration cytology (FNAC) samples. However, in a consecutive series of 40 FNAC samples only equivocal results were obtained on 38 benign and two malignant (FTC) thyroid tumour samples. CONCLUSIONS: Upregulation of ets-1 and downregulation of Tg mRNA expression occur in differentiated thyroid cancer and may facilitate pre operative identification of thyroid malignancy depending on further evaluation of these potentially promising markers in a larger series of benign and malignant thyroid tumours and their FNAC samples. PMID- 15879366 TI - Thyrotrophin-releasing hormone receptor 1 and prothyrotrophin-releasing hormone mRNA expression in the central nervous system are regulated by suckling in lactating rats. AB - BACKGROUND: The accepted function of the hypothalamic peptide, thyrotrophin releasing hormone (TRH), is to initiate release of thyrotrophin (TSH) from the pituitary. A physiological role for TRH in lactating rats has not yet been established. METHODS: Tissues were prepared from random-cycling and lactating rats and analysed using Northern blot, real time RT-PCR and quantitative in situ hybridisation. RESULTS: This study demonstrates that TRH receptor 1 (TRHR1) mRNA expression is up-regulated in the pituitary and in discrete nuclei of the hypothalamus in lactating rats, while proTRH mRNA expression levels are increased only in the hypothalamus. The results were corroborated by quantitative in situ analysis of proTRH and TRHR1. Bromocriptine, which reduced prolactin (PRL) concentrations in plasma of lactating and nursing rats, also counteracted the suckling-induced increase in TRHR1 mRNA expression in the hypothalamus, but had an opposite effect in the pituitary. These changes were confined to the hypothalamus and the amygdala in the brain. CONCLUSIONS: The present study shows that the mechanisms of suckling-induced lactation involve region-specific regulation of TRHR1 and proTRH mRNAs in the central nervous system notably at the hypothalamic level. The results demonstrate that continued suckling is critical to maintain plasma prolactin (PRL) levels as well as proTRH and TRHR1 mRNA expression in the hypothalamus. Increased plasma PRL levels may have a positive modulatory role on the proTRH/TRHR1 system during suckling. PMID- 15879367 TI - Systemic administration of pituitary adenylate cyclase-activating polypeptide maintains beta-cell mass and retards onset of hyperglycaemia in beta-cell specific calmodulin-overexpressing transgenic mice. AB - OBJECTIVE: Pituitary adenylate cyclase-activating polypeptide (PACAP) has been shown to play an important role in the regulation of islet function. We investigated its effects in beta-cell-specific calmodulin-overexpressing diabetic (CaMTg) mice, in which we consider that apoptosis of beta cells is the primary defect leading to basal hyperglycaemia. METHODS: CaMTg mice were treated with continuous s.c. infusions of PACAP from 2 to 4 weeks after birth, and were evaluated against littermate non-transgenic (nTg) and saline-treated CaMTg mice as to plasma glucose levels, insulin content, islet function and morphological features. RESULTS: Remarkable and progressive hyperglycaemia was observed in CaMTg mice, and PACAP treatment blunted this elevation. Insulin secretion from isolated islets demonstrated an impaired response to glucose in CaMTg mice, and PACAP treatment did not cause any improvement. The total pancreatic insulin content in CaMTg mice decreased significantly to 19.1% of that in nTg mice. PACAP treatment of CaMTg mice increased the content to 158% of the value in saline treated CaMTg mice. The insulin content in isolated islets from CaMTg mice also decreased to 15.9% of that in nTg mice, while PACAP treatment caused a doubling of the value. Immunohistochemical investigation revealed that the insulin positive islet area was markedly smaller in CaMTg mice and that PACAP treatment significantly expanded the insulin-positive islet area. CONCLUSIONS: These findings indicate that PACAP treatment retards the onset of hyperglycaemia in CaMTg mice by maintaining beta-cell mass and PACAP treatment may potentially be a therapeutic measure for preventing beta-cell exhaustion during hyperglycaemia. PMID- 15879368 TI - The polypill and cardiovascular disease. PMID- 15879369 TI - Pharmacogenetics and ethnically targeted therapies. PMID- 15879370 TI - Prevalence of asthma. PMID- 15879371 TI - Coping with the aftermath of trauma. PMID- 15879372 TI - Management of chronic kidney disease. PMID- 15879373 TI - Dutch approve euthanasia for a patient with Alzheimer's disease. PMID- 15879374 TI - Law lords give the go ahead for creation of "saviour siblings". PMID- 15879377 TI - Initiative could give free access to UK medical research. PMID- 15879381 TI - Legislation introduced to create a new drug safety office at the FDA. PMID- 15879380 TI - Doctor suspended as high death rate in emergency department is investigated. PMID- 15879382 TI - Doctor accused of exaggerating child sexual abuse. PMID- 15879383 TI - WHO combats counterfeit malaria drugs in Asia. PMID- 15879384 TI - GMC gives evidence to inquiry. PMID- 15879385 TI - People over 50 in northern Europe feel healthier than their southern counterparts. PMID- 15879387 TI - Obesity in children in England continues to rise. PMID- 15879388 TI - Trends in rates of different forms of diagnosed coronary heart disease, 1978 to 2000: prospective, population based study of British men. AB - OBJECTIVE: To examine trends over time in rates of different forms of diagnosed coronary heart disease among British men, during a period in which mortality due to coronary heart disease has been declining. DESIGN: Prospective cohort study covering the period 1978-80 to 1998-2000. PARTICIPANTS: 7735 men, aged 40-59 at entry, randomly selected from one general practice in each of 24 British towns. MAIN OUTCOME MEASURES: Trends in the rates of major coronary events, first diagnosed angina and first diagnosed coronary heart disease (any fatal or non fatal documented event or diagnosis). Events were ascertained from NHS central registers and reviews of medical records from general practices. RESULTS: Over the 20 year period, 1561 major coronary events occurred; 1087 and 1816 men had new diagnoses of angina and coronary heart disease, respectively. The age adjusted annual relative changes were -3.6% (95% confidence interval -4.8% to 2.4%, P < 0.001) for all major coronary events, 2.6% (1.1% to 4.0%, P < 0.001) for first diagnosed angina and -0.8% (-1.8% to 0.3%, P = 0.18) for first diagnosed coronary heart disease. The fall in major coronary events occurred across all categories of event (fatal and non-fatal, first and recurrent). Similarly, first diagnosed angina increased for both uncomplicated angina and angina after myocardial infarction. The age adjusted annual relative change in case fatality at 28 days of first major coronary events was -1.4% (-3.1% to 0.4%, P = 0.12). CONCLUSIONS: Among British middle aged men, a substantial decline in the rate of major coronary events over the past two decades seems to have been largely offset by an increase in the incidence of diagnosed angina. Overall there was little change in the incidence of first diagnosed coronary heart disease. A continuing need exists for resources and services for coronary heart disease in general, and for new angina in particular. PMID- 15879389 TI - Endorsement of the CONSORT statement by high impact medical journals: survey of instructions for authors. PMID- 15879390 TI - Effect of combinations of drugs on all cause mortality in patients with ischaemic heart disease: nested case-control analysis. AB - OBJECTIVE: To determine the effect of combinations of statins, aspirin, beta blockers, and angiotensin converting enzyme inhibitors in the secondary prevention of all cause mortality in patients with ischaemic heart disease. DESIGN: Open prospective cohort study with nested case-control analysis. SETTING: 1.18 million patients registered with 89 general practices across 23 strategic health authority areas within the United Kingdom. Practices had longitudinal data for a minimum of eight years and were contributing to QRESEARCH, a new database. PATIENTS: All patients with a first diagnosis of ischaemic heart disease between January 1996 and December 2003. Cases were patients with ischaemic heart disease who died. Controls were patients with ischaemic heart disease who were matched for age, sex, and year of diagnosis and were alive at the time their matched case died. MAIN OUTCOME MEASURES: Odds ratio with 95% confidence interval for risk of death in cases compared with controls. Exposure was current use of different combinations of statins, aspirin, beta blockers, and angiotensin converting enzyme inhibitors before death in cases, or the equivalent date in controls. RESULTS: 13,029 patients had a first diagnosis of ischaemic heart disease (incidence rate 338 per 100,000 person years). 2266 cases were matched to 9064 controls. Drug combinations associated with the greatest reduction in all cause mortality were statins, aspirin, and beta blockers (83% reduction, 95% confidence interval 77% to 88%); statins, aspirin, beta blockers, and angiotensin converting enzyme inhibitors (75% reduction, 65% to 82%); and statins, aspirin, and angiotensin converting enzyme inhibitors (71% reduction, 59% to 79%). Treatments associated with the smallest reduction in all cause mortality were beta blockers alone (19% reduction, 37% reduction to 4% increase), angiotensin converting enzyme inhibitors alone (20% reduction, 1% to 35%), and combined statins and angiotensin converting enzyme inhibitors (31% reduction, 57% reduction to 12% increase). CONCLUSIONS: Combinations of statins, aspirins, and beta blockers improve survival in high risk patients with cardiovascular disease, although the addition of an angiotensin converting enzyme inhibitor conferred no additional benefit despite the analysis being adjusted for congestive cardiac failure. PMID- 15879391 TI - Detecting cheating in written medical examinations by statistical analysis of similarity of answers: pilot study. AB - OBJECTIVE: To assess whether a computer program using a variant of Angoff's method can detect anomalous behaviour indicative of cheating in multiple choice medical examinations. DESIGN: Statistical analysis of 11 examinations held by the Royal College of Paediatrics and Child Health. SETTING: UK postgraduate medical examination. PARTICIPANTS: Examination candidates. MAIN OUTCOME MEASURES: Detection of anomalous candidate pairs by regression of similarity of correct answers in all possible pairs of candidates on the overall proportion of correct answers. Anomalous pairs were subsequently assessed in terms of examination centres and the seating plan of candidates, to assess adjacency. RESULTS: The 11 examinations were taken by a total of 11,518 candidates, and Acinonyx examined 6,178,628 pairs of candidates. Two examinations showed no anomalies, and one examination found an anomaly resulting from a scanning error. The other eight examinations showed 13 anomalies compatible with cheating, and in each pair the two candidates had sat the examination at the same centre, and for six examinations with seating plans, the candidates in the anomalous pairs had been seated side by side. The raw probabilities of the anomalies varied from 3.9x10( 11) to 9.3x10(-30) (median = 1.1x10(-17)), with Bonferroni-corrected probabilities in the range 2.4x10(-5) to 4.1x10(-24) (median = 1.6x10(-11)). This suggests that one anomalous pair is found for every 1000 or so candidates taking this postgraduate examination. CONCLUSIONS: This statistical technique identified a small proportion of candidates who had very similar patterns of correctly answered questions. The likelihood is that one candidate has copied from the other, or that there was collusion, or that a technical error occurred in the exams department (as happened in a single case). Analysis of similarities can be used to identify cheating and as part of the quality assurance process of postgraduate medical examinations. PMID- 15879393 TI - Postoperative hypoxia in a woman with Down's syndrome: case outcome. PMID- 15879394 TI - Doing what's best and best interests. PMID- 15879396 TI - Easy yet so easily missed. PMID- 15879397 TI - Quality assuring our learning from others. PMID- 15879398 TI - Underwater birth and neonatal respiratory distress. PMID- 15879399 TI - Hand. PMID- 15879400 TI - What can mendelian randomisation tell us about modifiable behavioural and environmental exposures? PMID- 15879401 TI - Why clinicians are natural bayesians. PMID- 15879402 TI - What's the evidence that NICE guidance has been implemented? Analysis is subject to confounding. PMID- 15879403 TI - Nurse led care: nurses are autonomous professionals delivering expert care. PMID- 15879405 TI - What's the evidence that NICE guidance has been implemented? Maybe NICE needs to do more to ensure implementation of guidelines. PMID- 15879406 TI - Influence of guidelines in determining medical negligence. PMID- 15879407 TI - What's the evidence that NICE guidance has been implemented? More recent data on NICE implementation show different picture. PMID- 15879408 TI - Long term sickness absence: general practitioners have crucial role but need political support. PMID- 15879409 TI - Long term sickness absence: motivation rather than ability to work may be key. PMID- 15879411 TI - Sit up and take notice about avian flu. PMID- 15879410 TI - Long term sickness absence: sickness absence, stress, and disaffection are linked. PMID- 15879412 TI - Neurotrophin overexpression in lower airways of infants with respiratory syncytial virus infection. AB - RATIONALE: Nerve growth factor and its receptors are upregulated in the lungs of weanling rats with lower respiratory tract infection caused by the respiratory syncytial virus (RSV), and this is a major mechanism of the inflammatory response against the virus. However, no information is available in humans. OBJECTIVES: We sought to determine whether the expression of neurotrophic factors and receptors is increased in human infants infected with RSV. METHODS: We used a highly sensitive immunoassay to measure the concentrations of nerve growth factor and brain-derived neurotrophic factor proteins in serum samples and in the supernatants and cell fractions of nonbronchoscopic bronchoalveolar lavage fluid. We also used immunofluorescence to detect expression in airway cells of the tyrosine kinase receptor TrkA, which binds nerve growth factor with high affinity. Samples were obtained at 24 and 48 hours postintubation from 31 mechanically ventilated infants: 15 patients with RSV infection, 5 patients with respiratory infection negative for RSV, and 11 postsurgical patients without respiratory infection. MAIN RESULTS: Data show significantly higher concentrations of both neurotrophins in the cell fractions of bronchoalveolar lavage samples obtained from infants with RSV infection compared with control infants, whereas no significant difference was found in supernatants or serum samples. We also detected tyrosine kinase receptor immunoreactivity only in cells from airways infected with RSV. CONCLUSIONS: We conclude that neurotrophic factors and receptors are overexpressed in human airways infected by RSV, and may contribute to airway inflammation and hyperreactivity. PMID- 15879413 TI - Decreased exhaled nitric oxide in pulmonary arterial hypertension: response to bosentan therapy. AB - RATIONALE: Decreased nitric oxide (NO) is considered an important pathogenetic mechanism in pulmonary arterial hypertension (PAH), but clear evidence is lacking. OBJECTIVES: We used multiple techniques to assess endogenous NO in 10 patients with untreated PAH (8 idiopathic and 2 anorexigen-associated PAH) and 12 control subjects. METHODS: After a nitrite/nitrate-restricted diet, NO metabolites (NOx) were assayed in 24-hour urine collections and exhaled NO (FE(NO)) determined at multiple expiratory flows. Analysis of the relation between FE(NO) and flow allowed derivation of three flow-independent parameters: airway wall concentration (C(W)), diffusing capacity (D(NO)), and alveolar concentration (C(A)). Seven patients underwent follow-up testing after 3 months of bosentan treatment. RESULTS: At baseline, FE(NO) was markedly decreased at the two lowest expiratory flows in PAH: 21 +/- 4 versus 36 +/- 4 ppb at 18 ml/second and 11 +/- 2 versus 17 +/- 2 ppb at 50 ml/second, for subjects with PAH and control subjects, respectively (p < 0.05). C(W) was 33 +/- 11 ppb in subjects with PAH versus 104 +/- 34 in control subjects (p = 0.04). Urinary NOx was also reduced in PAH (42 +/- 6 microM NOx/mM creatinine versus 62 +/- 7 in control subjects; p = 0.04). After bosentan, FE(NO), C(W), and urine NOx increased to control values (p < 0.05). Exclusion of the two anorexigen cases did not alter these results. CONCLUSIONS: FE(NO) at low expiratory flows was decreased in PAH due to reduced C(W). Bosentan reversed these abnormalities, suggesting that suppression of NO in PAH may have been caused by endothelin. PMID- 15879414 TI - A disintegrin and metalloprotease 33 polymorphisms and lung function decline in the general population. AB - RATIONALE: A disintegrin and metalloprotease 33 (ADAM33) has been identified as a susceptibility gene for asthma and single nucleotide polymorphisms (SNPs) in this gene have been associated with excessive decline of lung function in individuals with asthma. OBJECTIVES: To assess whether SNPs in ADAM33 are associated with accelerated lung function loss in the general population and with chronic obstructive pulmonary disease (COPD). METHODS: DNA was collected from subjects of the Vlagtwedde-Vlaardingen cohort participating in the last survey in 1989-1990 after a follow-up of 25 years. Information was collected every 3 years, including lung function measurements. We defined COPD as GOLD stage 2 or higher at the last survey. A total of 1,390 subjects from the cohort was genotyped for the following SNPs in ADAM33: F+1, Q-1, S_1, S_2, T_1, T_2, V_4, and ST+5. Differences in prevalence of SNPs were analyzed with chi(2) tests. Linear mixed effects models were used to analyze FEV(1) decline according to genotype. MEASUREMENTS AND MAIN RESULTS: In the whole population, mean adjusted decline was 18.7 and 12.7 ml/year in females and males, respectively. Individuals homozygous for minor alleles of SNPs S_2 and Q-1 and heterozygous for SNP S_1 had a significantly accelerated decline in FEV(1) of, respectively, 4.9, 9.6, and 3.6 ml/year compared with wild type. We found a significantly higher prevalence of SNPs F+1, S_1, S_2, and T_2 in subjects with COPD. CONCLUSIONS: We demonstrated that SNPs in ADAM33 are associated with accelerated lung function decline in the general population. These SNPs are also risk factors for COPD. PMID- 15879415 TI - Pulmonary dendritic cells. AB - Dendritic cells (DCs) are leukocytes that are emerging as chief orchestrators of immune responses. The crucial task of DCs is the continuous surveillance of antigen-exposed sites throughout the body, and their unique responsibility is to decide whether to present sampled antigen in an immunogenic or tolerogenic way. Any misstep can either lead to a flawed immune defense or to allergy, even autoimmunity. It comes as no surprise that the lungs become increasingly the subject of DC-related investigations, as they represent a vast interface between the body and the outer world. This constitutes an enormous challenge for the immune system: "firing up" immune responses inappropriately could have devastating results for the fragile gas exchange structures. Evidence accumulates that DCs play a pivotal role in maintaining the delicate balance between tolerance and active immune response in our respiratory system. The exponentially growing body of DC-related publications is a big challenge. This article aims to provide researchers and clinicians with an up-to-date view on DC biology and its relevance to pulmonary medicine. A developing trend in the field of DCs is the shift from fundamental immunologic research toward exciting clinical insights and applications. For the pulmonary clinician, this heralds the dawn of promising therapies in various domains such as infections, allergy, and cancer. PMID- 15879416 TI - CD14 tobacco gene-environment interaction modifies asthma severity and immunoglobulin E levels in Latinos with asthma. AB - BACKGROUND: A recent family-based genomewide screen revealed linkage between the 5q31 region and the diagnosis of asthma, but only in those exposed to environmental tobacco smoke (ETS). Among the candidate genes in this region is CD14. METHODS: To determine whether polymorphisms in the CD14 gene are related to this gene-by-environment interaction in Latinos, we used both family-based and cross-sectional cohort analysis to test for interactions between CD14 genotypes/haplotypes, exposure to ETS, and asthma-related phenotypes in 659 Mexican and Puerto Rican families. RESULTS: We identified 21 single nucleotide polymorphisms (SNPs) in the CD14 gene by sequencing 72 Puerto Ricans, Mexicans, and African Americans with asthma. Three SNPs, -810, -159, and +1437, were further genotyped in families with asthma. Among all subjects with asthma exposed to ETS, without regard to ethnicity, CD14 +1437 genotypes were associated with asthma severity. SNP +1437 GG or GC genotypes were significantly associated with lower baseline FEV1 using both family-based (p = 0.0009) and cross-sectional cohort (p = 0.03) analyses. Subjects with asthma with the GG or GC genotypes who were exposed to ETS had mean baseline FEV1 (% predicted) values 8.6% lower than subjects not exposed to ETS (p = 0.03). As previously observed in whites, we found an interaction between plasma IgE levels, SNP -159 genotypes, and ETS exposure (p = 0.0002). The lowest IgE levels were in those subjects with the TT genotype and who were exposed to ETS regardless of ethnicity. CONCLUSIONS: Our data suggest a gene-by-environment interaction between CD14 genotypes and ETS, which affects pulmonary function and IgE levels among Latinos with asthma. PMID- 15879417 TI - Positional identification of an asthma susceptibility gene on human chromosome 5q33. AB - RATIONALE: Asthma is a common respiratory disease with complex genetic components. We previously reported strong evidence for linkage between mite sensitive asthma and markers on chromosome 5q33. This area of linkage includes a region homologous to a mouse area that contains a locus involved in regulation of airway hyperreactivity. OBJECTIVE: The aim of the present study is to identify asthma susceptibility genes on chromosome 5q33. METHODS AND RESULTS: We performed mutation screening and association analyses of genes in the 9.4-Mb human linkage region. Transmission disequilibrium test analysis of 105 polymorphisms in 155 families with asthma revealed that six polymorphisms in cytoplasmic fragile X mental retardation protein (FMRP)-interacting protein 2 gene were associated significantly with the development of asthma (p = 0.000075; odds ratio, 5.9). These six polymorphisms were in complete linkage disequilibrium. In real-time quantitative polymerase chain reaction analysis, subjects homozygous for the haplotype overtransmitted to asthma-affected offspring showed significantly increased level of cytoplasmic FMRP interacting protein 2 gene expression in lymphocytes compared with ones heterozygous for the haplotype (p = 0.038). CONCLUSIONS: Our data suggest that cytoplasmic FMRP interacting protein 2 are associated with the development of atopic asthma in humans, and that targeting cytoplasmic FMRP interacting protein 2 could be a novel strategy for treating atopic asthma. PMID- 15879418 TI - Differential desensitization of homozygous haplotypes of the beta2-adrenergic receptor in lymphocytes. AB - Single-nucleotide polymorphisms of the beta(2)-adrenergic receptor gene and its 5' promoter have been associated with differences in receptor function and desensitization. Linkage disequilibrium may account for inconsistencies in reported effects of isolated polymorphisms. Therefore, we have investigated the three most common homozygous haplotypes of the beta(2)-adrenergic receptor (position 19 [Cys/Arg] of the 5' leader cistron and positions 16 [Arg/Gly] and 27 [Gln/Glu] of the receptor) for putative differences in agonist-induced desensitization. Lymphocytes of well defined nonasthmatic, nonallergic subjects homozygous for the haplotype CysGlyGln, ArgGlyGlu, or CysArgGln were isolated. Desensitization of (-)-isoproterenol-induced cyclic adenosine monophosphate (cAMP) accumulation and beta(2)-adrenergic receptor sequestration and downregulation were measured in relation to beta(2)-adrenergic receptor-mediated inhibition of IFN-gamma and interleukin-5 production. We observed that lymphocytes of individuals bearing the CysGlyGln haplotype were more susceptible to desensitization of the beta-agonist-induced cAMP response than those of individuals with the ArgGlyGlu or CysArgGln haplotype. The haplotype-dependent desensitization of beta-agonist-induced cAMP response was not associated with haplotype-dependent beta(2)-adrenergic receptor sequestration or downregulation. In addition, our data suggest reduced inhibition, in lymphocytes of subjects with the CysGlyGln haplotype, of interleukin-5 production induced by treatment with antibodies to the T-cell receptor-CD3 complex and to costimulatory molecule CD28 (alphaCD3/alphaCD28). This is the first study demonstrating haplotype-related differences in agonist-induced beta(2)-adrenergic receptor desensitization in primary human cells. This haplotype-related desensitization of the beta(2) adrenergic receptor in lymphocytes might have consequences regarding the regulation of helper T-cell type 2 inflammatory responses. PMID- 15879419 TI - Leukotriene modifier therapy for mild sleep-disordered breathing in children. AB - BACKGROUND: Children with mild sleep-disordered breathing (SDB), who may not be recommended for adenotonsillectomy, frequently exhibit neurocognitive and behavioral morbidity, and may benefit from alternative therapeutic interventions, such as leukotriene modifier therapy. METHODS: Twenty-four children with SDB completed an open-label intervention study for 16 weeks with daily montelukast therapy. Sleep studies and adenoid size estimates from lateral X-ray films of the neck were obtained before and after treatment. In a parallel study, adenoid and tonsillar tissues from children with obstructive sleep apnea or recurrent throat infections were subjected to quantitative polymerase chain reaction, immunohistochemistry, and Western blotting for gene and protein expression of leukotriene receptors LT1-R and LT2-R, and for concentrations of LTB4 and LTC4/D4/E4. RESULTS: Montelukast treatment induced significant reductions in adenoid size and respiratory-related sleep disturbances, which were absent in 16 children with SDB who did not receive treatment. LT1-R and LT2-R mRNA was similarly abundant in adenoid tissues, but increased LT1-R and LT2-R protein expression and higher levels of LTB4 and LTC4/D4/E4 emerged in children with obstructive sleep apnea. CONCLUSIONS: Oral therapy with a leukotriene modifier appears to be associated with improved breathing during sleep. Double-blind, placebo-controlled trials will be needed to corroborate current findings and solidly establish antiinflammatory strategies, such as leukotriene modifiers, as therapeutic alternatives in children with SDB too mild to justify referral for adenotonsillectomy. PMID- 15879420 TI - Clinical advances in the diagnosis and therapy of the interstitial lung diseases. AB - The last century experienced remarkable advances in the classification, diagnosis, and understanding of the pathogenesis of the interstitial lung diseases. Technological advances, particularly physiologic testing, lung imaging studies, bronchoalveolar lavage, surgical lung biopsy, and histopathologic assessment, improved our understanding of these entities. In particular, the advent of high-resolution computed tomography, the narrowed pathologic definition of usual interstitial pneumonia, and recognition of the prognostic importance of separating usual interstitial pneumonia from other idiopathic interstitial pneumonia patterns have profoundly changed the approach to these processes. Most recently, genetic medicine, the use of new technologies (e.g., microarrays, mass spectroscopic analysis of proteins, and laser capture microdissection), and the development of animal models have had a major impact on understanding the pathogenesis and potential molecular targets for interfering with fibrogenesis. This article highlights some of the advances and changes in clinical practice that took place in the management of patients with interstitial lung diseases over the last century. PMID- 15879421 TI - Inhaled iloprost reverses vascular remodeling in chronic experimental pulmonary hypertension. AB - RATIONALE: Inhaled iloprost is an effective therapy for pulmonary arterial hypertension (PAH). However, no study to date has addressed the effects of inhaled iloprost on changes to pulmonary vascular structure that occur in PAH. OBJECTIVES: The present study was designed to investigate chronic antiremodeling effects of inhaled iloprost in monocrotaline (MCT)-induced PAH in rats. METHODS: Four weeks after a single injection of MCT, after full establishment of PAH, rats were nebulized with iloprost at a dose of 6 microg . kg(-1) . day(-1), or underwent sham nebulization with saline. RESULTS: After 2 weeks of inhalation therapy, right ventricular pressure and pulmonary vascular resistance were reversed in rats treated with iloprost, but not in sham-treated control animals. Systemic arterial pressure was unaffected. In addition, right heart hypertrophy, the degree of pulmonary artery muscularization, and the medial wall thickness of intraacinar pulmonary arteries regressed in response to iloprost. Furthermore, the MCT-induced increase in matrix metalloproteinase-2 and -9 activities and tenascin-C expression was suppressed. CONCLUSIONS: We conclude that the inhalation of iloprost reverses PAH and vascular structural remodeling in MCT treated rats. This regimen suggests the possibility of an antiremodeling therapy in PAH. PMID- 15879422 TI - House dust mite facilitates ovalbumin-specific allergic sensitization and airway inflammation. AB - RATIONALE: Mouse models of allergic airway disease have greatly contributed to our understanding of disease induction and pathogenesis. Although these models typically investigate responses to a single antigen or allergen, humans are frequently exposed to a myriad of allergens, each with distinct antigenic potential. OBJECTIVES: Given that airway exposure to ovalbumin (OVA), a prototypic innocuous antigen, induces inhalation tolerance, we wished to investigate how this response would be altered if OVA were encountered concurrently with a house dust mite extract (HDM), which we have recently shown is capable of eliciting a robust allergic airway inflammatory response that is mediated, at least in part, by granulocyte-macrophage colony-stimulating factor. METHODS: Balb/c mice were exposed daily to HDM (intranasally) followed immediately by exposure to aerosolized OVA for 5 weeks. To allow the inflammatory response elicited by HDM to subside fully, mice were then allowed to rest, unexposed, for 8 weeks, at which time they were rechallenged with aerosolized OVA for 3 consecutive days. MEASUREMENTS AND MAIN RESULTS: At this time, we observed a robust eosinophilic inflammatory response in the lung that was associated with an increase in bronchial hyperreactivity. Moreover, we documented significantly elevated serum levels of OVA-specific IgE and IgG(1) and increased production of the Th2 cytokines interleukin 4 (IL-4), IL-5, and IL-13 by splenocytes stimulated in vitro with OVA. CONCLUSION: Our data demonstrate the potential of a potent allergen such as HDM to establish a lung microenvironment that fosters the development of allergic sensitization to otherwise weak or innocuous antigens, such as OVA. PMID- 15879423 TI - Surfactant palmitoylmyristoylphosphatidylcholine is a marker for alveolar size during disease. AB - Two common lung-related complications in the neonate are respiratory distress syndrome, which is associated with a failure to generate low surface tension at the air-liquid interface because of pulmonary surfactant insufficiency, and bronchopulmonary dysplasia (BPD), a chronic lung injury with reduced alveolarization. Surfactant phosphatidylcholine (PC) molecular species composition during alveolarization has not been examined. Mass spectrometry analysis of bronchoalveolar lavage fluid of rodents and humans revealed significant changes in surfactant PC during alveolar development and BPD. In rats, total PC content rose during alveolarization, which was caused by an increase in palmitoylmyristoyl-PC (16:0/14:0PC) concentration. Furthermore, two animal models of BPD exhibited a specific reduction in 16:0/14:0PC content. In humans, 16:0/14:0PC content was specifically decreased in patients with BPD and emphysema compared with patients without alveolar pathology. Palmitoylmyristoyl PC content increased with increasing intrinsic surfactant curvature, suggesting that it affects surfactant function in the septating lung. The changes in acyl composition of PC were attributed to type II cells producing an altered surfactant during alveolar development. These data are compatible with extracellular surfactant 16:0/14:0PC content being an indicator of alveolar architecture of the lung. PMID- 15879424 TI - Inhaled corticosteroid dose reduction in childhood asthma: is nitrosopnea informative? PMID- 15879425 TI - Haplotypes and asthma. PMID- 15879426 TI - Tobacco money at the University of California. PMID- 15879427 TI - The CXCL10/CXCR3 axis mediates human lung mast cell migration to asthmatic airway smooth muscle. AB - Mast cell microlocalization within the airway smooth muscle bundle is an important determinant of the asthmatic phenotype. We hypothesized that mast cells migrate toward airway smooth muscle in response to smooth muscle-derived chemokines. In this study, we investigated (1) chemokine receptor expression by mast cells in the airway smooth muscle bundle in bronchial biopsies from subjects with asthma using immunohistology, (2) the concentration of chemokines in supernatants from stimulated ex vivo airway smooth muscle cells from subjects with and without asthma measured by enzyme-linked immunosorbent assay, and (3) mast cell migration toward these supernatants using chemotaxis assays. We found that CXCR3 was the most abundantly expressed chemokine receptor on human lung mast cells in the airway smooth muscle in asthma and was expressed by 100% of these mast cells compared with 47% of mast cells in the submucosa. Human lung mast cell migration was induced by airway smooth muscle cultures predominantly through activation of CXCR3. Most importantly, CXCL10 was expressed preferentially by asthmatic airway smooth muscle in bronchial biopsies and ex vivo cells compared with those from healthy control subjects. These results suggest that inhibition of the CXCL10/CXCR3 axis offers a novel target for the treatment of asthma. PMID- 15879429 TI - Fluticasone reduces CRP in COPD. PMID- 15879430 TI - Systemic bioavailability of fluticasone in COPD. PMID- 15879431 TI - Post-translational modification of nuclear co-repressor receptor-interacting protein 140 by acetylation. AB - Receptor-interacting protein 140 (RIP140) is a versatile co-regulator for nuclear receptors and many transcription factors and contains several autonomous repressive domains. RIP140 can be acetylated, and acetylation affects its biological activity. In this study, a comprehensive proteomic analysis using liquid chromatography-tandem mass spectroscopy was conducted to identify the in vivo acetylation sites on RIP140 purified from Sf21 insect cells. Eight acetylation sites were found within the amino-terminal and the central regions, including Lys111, Lys158, Lys287, Lys311, Lys482, Lys529, Lys607, and Lys932. Reporter assays were conducted to examine the effects of acetylation on various domains of RIP140. Green fluorescent protein-tagged fusion proteins were used to demonstrate the effect on nuclear translocation of these domains. A general inhibitor of reversible protein deacetylation was used to enrich the acetylated population of RIP140. The amino-terminal region (amino acids (aa) 1-495) was more repressive and accumulated more in the nuclei under hyperacetylated conditions, whereas hyperacetylation reduced the repressive activity and nuclear translocation of the central region (aa 336-1006). The deacetylase inhibitor had no effect on the carboxyl-terminal region (aa 977-1161) where no acetylation sites were found. Hyperacetylation also enhanced the repressive activity of the full-length protein but triggered its export into the cytosol in a small population of cells. This study revealed differential effects of post translational modification on various domains of RIP140 through acetylation, including its effects on repressive activity and nuclear translocation of the full-length protein and its subdomains. PMID- 15879432 TI - Quantification of gel-separated proteins and their phosphorylation sites by LC-MS using unlabeled internal standards: analysis of phosphoprotein dynamics in a B cell lymphoma cell line. AB - Protein phosphorylation plays a critical role in normal cellular function and is often subverted in disease. Although major advances have recently been made in identification and quantitation of protein phosphorylation sites by MS, current methodological limitations still preclude routine, easily usable, and comprehensive quantitative analysis of protein phosphorylation. Here we report a simple LC-MS method to quantify gel-separated proteins and their sites of phosphorylation; in this approach, integrated chromatographic peak areas of peptide analytes from proteins under study are normalized to those of a non isotopically labeled internal standard protein spiked into the excised gel samples just prior to in-gel digestion. The internal standard intensities correct for differences in enzymatic activities and sample losses that may occur during the processes of in-gel digestion and peptide extraction from the gel pieces. We used this method of peak area measurement with an internal standard to investigate the effects of pervanadate on protein phosphorylation in the WEHI-231 B cell lymphoma cell line and to assess the role of phosphoinositide 3-kinase (PI3K) in these phosphorylation events. Phosphoproteins, isolated from total cell lysates using IMAC or by immunoprecipitation using Tyr(P) antibodies, were analyzed using this method, leading to identification of >400 proteins, several of which were found at higher levels in phosphoprotein fractions after pervanadate treatment. Pretreatment of cells with the PI3K inhibitor wortmannin reduced the phosphorylation level of certain proteins (e.g. STAT1 and phospholipase Cgamma2) while increasing the phosphorylation of several others. Peak area measurement with an internal standard was also used to follow the dynamics of PI3K-dependent and -independent changes in the post-translational modification of both known and novel phospholipase Cgamma2 phosphorylation sites. Our results illustrate the capacity of this conceptually simple LC-MS method for quantification of gel-separated proteins and their phosphorylation sites and for quantitative profiling of biological systems. PMID- 15879433 TI - Haplotype analysis indicates an association between the DOPA decarboxylase (DDC) gene and nicotine dependence. AB - DOPA decarboxylase (DDC; also known as L-amino acid decarboxylase; AADC) is involved in the synthesis of dopamine, norepinephrine and serotonin. Because the mesolimbic dopaminergic system is implicated in the reinforcing effects of many drugs, including nicotine, the DDC gene is considered a plausible candidate for involvement in the development of vulnerability to nicotine dependence (ND). Further, this gene is located within the 7p11 region that showed a 'suggestive linkage' to ND in our previous genome-wide scan in the Framingham Heart Study population. In the present study, we tested eight single nucleotide polymorphisms (SNPs) within DDC for association with ND, which was assessed by smoking quantity (SQ), the heaviness of smoking index (HSI) and the Fagerstrom test for ND (FTND) score, in a total of 2037 smokers and non-smokers from 602 nuclear families of African- or European-American (AA or EA, respectively) ancestry. Association analysis for individual SNPs using the PBAT-GEE program indicated that SNP rs921451 was significantly associated with two of the three adjusted ND measures in the EA sample (P=0.01-0.04). Haplotype-based association analysis revealed a protective T-G-T-G haplotype for rs921451-rs3735273-rs1451371-rs2060762 in the AA sample, which was significantly associated with all three adjusted ND measures after correction for multiple testing (min Z=-2.78, P=0.006 for HSI). In contrast, we found a high-risk T-G-T-G haplotype for a different SNP combination in the EA sample, rs921451-rs3735273-rs1451371-rs3757472, which showed a significant association after Bonferroni correction with the SQ and FTND score (max Z=2.73, P=0.005 for FTND). In summary, our findings provide the first evidence for the involvement of DDC in the susceptibility to ND and, further, reveal the racial specificity of its impact. PMID- 15879434 TI - Impaired cotranslational processing of the calcium-sensing receptor due to signal peptide missense mutations in familial hypocalciuric hypercalcemia. AB - The CASR, a cell surface glycoprotein expressed in parathyroid gland and kidney, is critical for maintaining extracellular calcium homeostasis. The inherited disorders, familial hypocalciuric hypercalcemia (FHH) and neonatal severe hyperparathyroidism (NSHPT), are caused by inactivating mutations in the CASR gene. The CASR has an N-terminal, 19 amino acid signal peptide that is predicted to direct the nascent polypeptide chain, as it emerges from the ribosome, into the endoplasmic reticulum (ER). Here, we report the functional characterization of three CASR mutations identified in hypercalcemic/hyperparathyroid patients. The mutations, L11S, L13P and T14A, lie within the signal peptide hydrophobic core. When transiently transfected into kidney cells, L11S and L13P mutants demonstrated reduced intracellular and plasma membrane expression and signaling to the mitogen-activated protein kinase pathway in response to extracellular calcium relative to wild-type CASR and the T14A mutant. All mutant CASR RNAs translated into protein normally. In cotranslational processing assays, which test the functionality of the signal peptide in the early secretory pathway, the wild-type CASR and mutant T14A nascent polypeptides were targeted to microsomal vesicles, representing the ER, translocated into the vesicular lumen and underwent core N-glycosylation. In contrast, the L11S and L13P mutants failed to be inserted in the microsomes and undergo glycosylation. This is the first study examining the function of the CASR signal sequence and reveals that both L11S and L13P mutants are markedly impaired with respect to cotranslational processing, accounting for the observed parathyroid dysfunction. PMID- 15879435 TI - Molecular properties and pharmacogenetics of a polymorphism of adenylyl cyclase type 9 in asthma: interaction between beta-agonist and corticosteroid pathways. AB - In asthma, the response to beta-agonists acting at beta2-adrenergic receptors (beta2AR) displays extensive interindividual variation. One effector for airway beta2AR, adenylyl cyclase type 9 (AC9), was considered a candidate locus for predicting beta-agonist efficacy in the absence and presence of corticosteroid treatment. One non-synonymous AC9 polymorphism has been identified, which results in substitution of Met for Ile at amino acid 772. Under standard culture conditions in stably transfected cells, we found decreased catalytic activity of Met772. However, cells cultured in the presence of glucocorticoid expressing Met772 had a significantly increased albuterol-stimulated adenylyl cyclase response (approximately 80%) when compared with those expressing Ile772 (approximately 20%, P=0.02). An equivalent increase in beta2AR expression was observed in both lines due to glucocorticoid, but AC9 expression was unaffected. The hypothesis that Met772-AC9 is associated with an improved albuterol bronchodilator response in asthmatics was investigated in 436 asthmatic children who were followed for 4 years and randomized to receive placebo or the inhaled corticosteroid budesonide. Met772 carriers on budesonide showed a significant improvement in forced expiratory volume in 1 s (P=0.005). Moreover, a highly significant interaction (P=0.002) was found for budesonide treatment and the AC9 polymorphism. These in vitro and human association studies are consistent with this AC9 polymorphism altering albuterol responsiveness in the context of concomitant inhaled corticosteroid administration, which is a common asthma regimen. The Met772-AC9 polymorphism represents one of most likely several multi gene polymorphisms along the receptor-relaxation axis, which together may provide for a composite pharmacogenetic index for asthma therapy. PMID- 15879436 TI - HspB8, a small heat shock protein mutated in human neuromuscular disorders, has in vivo chaperone activity in cultured cells. AB - The family of small heat shock proteins (sHsp) is composed of 10 members in mammals, four of which are found mutated in diseases associated with the accumulation of protein aggregates. Though many sHsp have demonstrated molecular chaperone activity in vitro in cell-free conditions, their activity in vivo in the normal cellular context remains unclear. In the present study, we investigated the capacity of the sHsp, HspB8/Hsp22, to prevent protein aggregation in the cells using the polyglutamine protein Htt43Q as a model. In control conditions, Htt43Q accumulated in perinuclear inclusions composed of SDS insoluble aggregates. Co-transfected with Htt43Q, HspB8 became occasionally trapped within the inclusions; however, in most cells, HspB8 blocked inclusion formation. Biochemical analyses indicated that HspB8 inhibited the accumulation of SDS-insoluble Htt43Q as efficiently as Hsp40 which was taken as a positive control. Htt43Q then accumulated in the SDS-soluble fraction, provided that protein degradation was blocked by proteasome and autophagy inhibitors. In contrast, the other sHsp Hsp27/HspB1 and alphaB-crystallin/HspB5 had no effect. This suggested that HspB8 functions as a molecular chaperone, maintaining Htt43Q in a soluble state competent for rapid degradation. Analyses of Hsp27-HspB8 chimeric proteins indicated that the C-terminal domain of HspB8 contains the specific sequence necessary for chaperone activity. Missense mutations in this domain at lysine 141, which are found in human motor neuropathies, significantly reduced the chaperone activity of the protein. A decrease in the HspB8 chaperone activity may therefore contribute to the development of these diseases. PMID- 15879437 TI - Polymorphisms in the FCN2 gene determine serum variation and function of Ficolin 2. AB - The ficolin 1, 2 and 3 (derived from the FCN1, 2 and 3 genes, respectively) are homologous soluble pattern recognition molecules of importance for innate immunity, comprising collagen-like and fibrinogen-like domains, binding to sugar groups on different types of microorganisms. Serum concentration of Ficolin-2 varies considerably in healthy individuals. Thus, we speculated whether this could be due to variations in the FCN2 gene. We sequenced the promoter region and the exons and intron-exon boundaries of FCN2 in Danish Caucasians. For comparison, FCN1 and FCN3 were also investigated. Ficolin-2 concentrations were measured in serum and the functional relevance of amino acid substituting polymorphisms in FCN2 was investigated by binding to and recovery from N acetylglucosamine (GlcNAc). Both FCN1 and FCN2 contained polymorphisms in the promoters and structural parts of the genes, but only polymorphisms in FCN2 resulted in amino acid exchanges. FCN2 promoter polymorphisms were associated with marked changes in the Ficolin-2 serum concentration, whereas two polymorphisms clustered in the exon encoding the fibrinogen-like domain were associated with increased and decreased GlcNAc binding, respectively. In FCN3, only a single frame-shift deletion in exon 5 was detected. These results show that the FCN genes are polymorphic and that particularly FCN2 harbors functional polymorphic sites that regulate both the expression as well as the function of Ficolin-2, which may have pathophysiological implications for innate immunity. PMID- 15879438 TI - The effect of Prime Minister Anthony Eden's illness on his decision-making during the Suez crisis. PMID- 15879439 TI - Osteoporosis therapy: an example of putting evidence-based medicine into clinical practice. AB - A major aim of evidence-based medicine is to assist clinical decision-making by providing the most current and reliable medical information. Systematic reviews and meta-analyses are important tools in this process. Systematic reviews identify and compile relevant evidence, while meta-analyses summarize and quantify the results of such reviews. Results from meta-analyses are, at present, the main source of summary evidence for the efficacy of treatments for a specific condition. They are important tools for helping to choose among treatment options, although they cannot be used to directly compare the magnitude of the effect of various therapies. However, the methods used and the consequent clinical value of the results, may be poorly understood by clinicians, who may therefore not take full advantage of the evidence. Recently, a panel of experts in osteoporosis and evidence-based medicine applied rigorous, validated, scientific standards to produce a systematic review and meta-analysis of randomized controlled trials of anti-resorptive agents used to treat osteoporosis. They found that, although several agents reduced the risk of vertebral fracture, only two, alendronate and risedronate, demonstrated convincing evidence for both non-vertebral and vertebral fracture risk reductions. The clinical implication of these results is that there are important differences in anti-fracture efficacy among currently available agents. In the absence of evidence from head-to-head clinical trials and because of the systematic nature and methodological rigor of the analyses, these data provide important information for clinical decision-making. PMID- 15879440 TI - Faecal calprotectin in the assessment of Crohn's disease activity. AB - BACKGROUND: Clinical and laboratory assessment of activity in Crohn's disease (CD) correlate poorly with endoscopic findings. Calprotectin is a calcium-binding protein abundant in neutrophil cytosol, and extremely stable in faeces. Faecal calprotectin (FC) is an excellent surrogate marker of neutrophil influx into the bowel lumen. AIM: To assess whether FC concentration from a spot stool sample reliably detects active inflammation in patients with CD. DESIGN: Cross-sectional comparative study. METHODS: Subjects had a previously confirmed diagnosis of CD and were suspected on clinical grounds to be in the midst of a relapse. Thirty five entered the study; they underwent radiolabelled white cell scanning (WCS) and had a stool sample collected for calprotectin measurement on the same day. A Crohn's disease activity index (CDAI) was also calculated for each. The WCS scans were scored at six standard sites to give a mean total, 'extent', 'severity' and 'combined extent and severity' scores. RESULTS: FC was significantly and positively correlated with mean total (r = 0.73, p < 0.001), 'extent' (r = 0.71, p < 0.001), 'severity' (r = 0.64, p < 0.001) and combined 'extent and severity' WCS scores (r = 0.71, p < 0.001). A cut-off of faecal calprotectin > 100 microg/g gave a sensitivity of 80%, specificity of 67%, positive predictive value of 87% and a negative predictive value of 64% in identifying those with and without any inflammation on WCS. There was, however, no significant correlation between CDAI and mean total WCS score (r = 0.21, p = 0.24), nor between CDAI and FC (r = 0.33, p = 0.06). DISCUSSION: While the CDAI does not accurately reflect inflammatory activity in CD, a one-off FC reliably detects the presence or absence of intestinal inflammation in adult patients with CD, compared to WCS. PMID- 15879441 TI - Diazepam with your dinner, Sir? The lifestyle drug-substitution strategy: a radical alcohol policy. AB - Over recent decades the drink problem in the British Isles has grown to become arguably the worst in the Western world, combining the high average alcohol intake of southern Europe with binge-drinking typical of extreme latitudes. Since the problem continues to worsen, and traditional strategies regulating price and access are probably untenable, radical new alcohol policies are required. The drug-substitution strategy is based on an assumption that most people use lifestyle drugs rationally for self-medication purposes, to achieve specific desired psychological effects, especially enhanced well-being. When there is access to an equally effective, but safer, alternative drug, then people would tend to switch to it (especially when the substitute is legal and socially acceptable). There are several safer lifestyle drug-substitutes for alcohol, including benzodiazepines, SSRIs and marijuana. Southern Europeans use alcohol mainly as an anxiolytic social lubricant, taken in low but frequent doses with high annual per capita consumption, and for this pattern, benzodiazepines might be a medically safer lifestyle drug-substitute. Northern Europeans traditionally use alcohol in high doses as a euphoric intoxicant, and for this pattern, marijuana might be a safer and less-antisocial substitute. Since this risk benefit calculus implies that there are better alternatives to alcohol, government policy should promote safer lifestyle drug-substitutes by removing legal barriers and altering the balance of economic and social incentives. PMID- 15879442 TI - Hypertension-based clinical risk strategies for detecting microalbuminuria in diabetes. AB - BACKGROUND: Microalbuminuria screening to identify patients at risk of diabetic nephropathy is widely accepted. AIM: To investigate whether blood-pressure-based strategies can identify such patients without the need for microalbuminuria testing. METHODS: Spot urine for albumin/creatinine ratios was performed in all patients over an 18-month period. The performance of four combinations of clinical models, based on existing triggers for anti-hypertensive intervention (prior use and/or existing systolic BP exceeding 140 or 160 mmHg and/or dipstick proteinuria exceeding 1+ or 2+) was evaluated at microalbuminuria thresholds of 3.5 and 10 mg/mmol. The models were ranked 1 to 4, based on their escalating relative strengths in predicting need for intervention. RESULTS: Of 3748 patients, 1257 (34%) or 739 (20%) exceeded microalbuminuria thresholds of 3.5 or 10 mg/mmol. All four models predicted microalbuminuria risk (areas under ROC curves 0.60-0.77, all p < 0.001). The models (1-4) identified 2220, 2465, 2803 or 2937 for intervention, respectively, irrespective of microalbuminuria status, and missed 368, 232, 194 or 126 at 3.5 mg/mmol and 164, 87, 81 or 45 at 10 mg/mmol. DISCUSSION: Clinical models using routinely measured parameters reduced the target population for microalbuminuria screening by 60-80%, missing 3-10% of patients with albumin/creatinine ratios exceeding 3.5 mg/mmol or 1-4% of those exceeding 10 mg/mmol. PMID- 15879443 TI - Cost-effectiveness of integrated stroke services. AB - BACKGROUND: Randomized trials have shown that integrating services for acute stroke care may lead to organizational improvements, higher efficiency and better patient outcomes in the acute phase. AIM: To compare the costs and effects of stroke services in an experimental group of patients compared to a group of patients receiving conventional care. DESIGN: Prospective non-randomized controlled trial. METHODS: We compared all consecutively hospitalized stroke patients in three experimental stroke service settings (Delft, Haarlem and Nijmegen, n = 411) with concurrent patients receiving conventional stroke care (n = 187) over 6 months follow-up. Main end-points were total costs per patient and total health-adjusted days per 100 patients as measured by the EuroQol-5D score during follow-up. RESULTS: Mean total costs per patient were 16,000 Euro (95%CI 14,670 Euro-16,930 Euro): 13,160 Euro in Delft, 16,790 Euro in Haarlem, 20,230 Euro in Nijmegen, and 13,810 Euro in the control regions. Early discharge in Delft saved about 2500 Euro hospital costs per patient. General patient health in Delft was significantly better than in the control regions; Haarlem and Nijmegen showed no difference in health. DISCUSSION: Our study confirms the potential to improve stroke outcomes in a cost-effective way in Dutch settings. This was seen in the group of patients in Delft, a complete and relatively simple stroke service, but not in two other regions with more complex stroke services. Important factors are reduction of hospital days and, most likely, adequate multidisciplinary rehabilitation. PMID- 15879444 TI - Fungal infections as a complication of therapy for sarcoidosis. AB - BACKGROUND: Treatment of symptomatic sarcoidosis usually includes systemic immunosuppressive agents. These agents may render the patient more susceptible to opportunistic infections. In addition, the fungal infection may be difficult to distinguish from the underlying sarcoidosis. AIM: To examine the presentation and management of invasive fungal infections in sarcoidosis patients. DESIGN: Retrospective record review. METHODS: We reviewed the notes of all sarcoidosis patients (n = 753) seen at our clinic over an 18-month period. RESULTS: Seven patients (0.9%) with previously diagnosed sarcoidosis developed fungal infections: two each with Histoplasma capsulatum and Blastomyces dermatitidis and three others with Cryptococcus neoformans. No cases of invasive aspergillus or tuberculosis were identified. The diagnosis of fungal infection was made by bronchoscopy (four cases), open-lung biopsy (one case), bone-marrow aspirate (one case), and spinal fluid examination (one case). All patients were receiving corticosteroids at the time of worsening chest X-ray or clinical status. Four patients were also receiving methotrexate prior to infection. No patient with systemic fungal infection was receiving either infliximab or cyclophosphamide. All patients responded to anti-fungal therapy and a reduction in immunosuppression. DISCUSSION: Fungal infections occur rarely in treated patients with sarcoidosis. Deterioration of chest X-ray, especially a localized infiltrate, warrants investigation. PMID- 15879445 TI - Treatment and outcome of adult patients with primary focal segmental glomerulosclerosis in five UK renal units. AB - BACKGROUND: Focal segmental glomerulosclerosis (FSGS) is the least studied of the causes of idiopathic nephrotic syndrome, and there are few specific guidelines for treatment. AIM: To review data from five UK renal units to investigate whether adult patients with FSGS were treated uniformly, and to examine the effect of treatment on proteinuria and survival. DESIGN: Retrospective record review. METHODS: We examined electronic records of patients with idiopathic FSGS for information on baseline clinical parameters, treatment regimens and outcomes. RESULTS: Of 136 patients with primary FSGS and nephrotic range proteinuria, 76 (56%) were treated with prednisolone and of this group, 59% were treated with additional immunosuppression. Among the treated patients, the total remission rate (complete and partial) was 67%, and one hospital achieved a remission rate of 80%. Treated patients had a significantly higher remission rate than those who were not treated. Remission was associated with a 5-year survival off dialysis of 94%, compared with 53% if remission was not achieved. Baseline serum creatinine and remission were independently associated with survival off dialysis in a multivariate Cox proportional hazards model. DISCUSSION: Patients with primary FSGS and nephrotic range proteinuria, who are treated with corticosteroids, are more likely to enter remission than those who are not treated. Remission rates of up to 80% can be achieved with prolonged treatment, and remission is an independent predictor of survival off dialysis. Patients who do not achieve remission have a poor prognosis. Further clarification of optimal treatment regimens requires additional, prospective studies. PMID- 15879446 TI - Meaningful workplace protection factor measurement: experimental protocols and data treatment. AB - Workplace performance measurement of respiratory protective equipment (RPE) is fundamental to the understanding of how well wearers are protected. It forms the basis for guidance on which the selection of appropriate equipment is based. However, the measurement of this performance is open to many sources of interference and inaccuracy, reducing the value and relevance of the results, and is most difficult for devices providing the highest levels of protection. In this paper, a method for critically assessing collected workplace protection factor (WPF) data is validated. This method rejects unreliable data, using criteria based on the detection limits of the analytical measurement system. An iterative approach is also described which arrives at a supportable estimate of given non parametric percentiles of the distribution of measured WPFs [such as the fifth percentile, conventionally taken to be the assigned protection factor (APF)]. Further pragmatic criteria, based on a combination of experimental experience and consideration from first principles, are suggested as the basis for any future studies of RPE performance. These will maximize the chances of valid measurements being made, and also provide insight into the level of confidence which can be placed on any of the results. A consequence of these criteria is that typical working environments and measurement methods are incapable of generating WPF data which can be considered reliable. PMID- 15879447 TI - Volatile C6-aldehydes and Allo-ocimene activate defense genes and induce resistance against Botrytis cinerea in Arabidopsis thaliana. AB - Green leafy volatiles or isoprenoids are produced after mechanical wounding or pathogen/herbivore attacks in higher plants. We monitored expression profiles of the genes involved in defense responses upon exposing Arabidopsis thaliana to the volatiles. Among the genes investigated, those known to be induced by mechanical wounding and/or jasmonate application, such as chalcone synthase (CHS), caffeic acid-O-methyltransferase (COMT), diacylglycerol kinase1 (DGK1), glutathione-S transferase1 (GST1) and lipoxygenase2 (LOX2), were shown to be induced with (E)-2 hexenal, (Z)-3-hexenal, (Z)-3-hexenol or allo-ocimene (2,6-dimethyl-2,4,6 octatriene). A salicylic acid-responsive gene, pathogenesis-related protein2 (PR2), was not induced by the volatiles. Detailed analyses of the expression profiles showed that the manner of induction varied depending on either the gene monitored or the volatile used. A chemically inert compound, (Z)-3-hexenol, was also potent, which suggested that chemical reactivity was not the sole requisite for the inducing activity. With a jasmonate-insensitive mutant (jar1), the induction by the volatiles was mostly suppressed, however, that of LOX2 was unaltered. An ethylene-insensitive mutant (etr1) showed responses almost identical to the wild type, with minor exceptions. From these observations, it was suggested that both the jasmonate-dependent and -independent pathways were operative upon perception of the volatiles, while the ETR1-dependent pathway was not directly involved. When Botrytis cinerea was inoculated after the volatile treatment, retardation of disease development could be seen. It appears that volatile treatment could make the plants more resistant against the fungal disease. PMID- 15879448 TI - Involvement of ARM2 in the uptake of indole-3-butyric acid in rice (Oryza sativa L.) roots. AB - Auxin influx carriers are involved in auxin transport and plant development. Here we show that the mutant of rice (Oryza sativa L. ssp. indica cv IR8) arm2 is defective in the uptake of the naturally occurring auxin indole-3-butyric acid (IBA). The acropetal and basipetal transport of IBA is reduced in arm2 roots compared with wild type. In contrast, arm2 roots are normal with respect to uptake and transport of indole-3-acetic acid (IAA). Furthermore, arm2 roots are resistant to IBA but respond normally to IAA. The mutant analysis of arm2 indicates the presence of an influx carrier system for IBA in rice roots. PMID- 15879449 TI - Regulation of TOP2 by various abiotic stresses including cold and salinity in pea and transgenic tobacco plants. AB - Although abiotic stress affects plant growth and development, demonstration of its direct effect on regulation of the components of the DNA replication machinery is largely unknown. In this work, we demonstrate that the expression of TOP2 (encoding topoisomerase II) is up-regulated by various abiotic stresses including salinity and low temperature, and phytohormones such as ABA (abscisic acid) and SA (salicylic acid). Transgenic studies with different deletion versions of the TOP2 promoter in tobacco define several promoter determinants responsible for specific abiotic stress responsiveness. Taken together, these results demonstrate a direct involvement of stress in the transcriptional regulation of TOP2. PMID- 15879450 TI - CGHMultiArray: exact P-values for multi-array comparative genomic hybridization data. AB - We compute P-values, based on the Wilcoxon test with ties, to compare two conditions with array comparative genomic hybridization data, and we provide a simple interface to export and plot these P-values. PMID- 15879451 TI - Fast parsers for Entrez Gene. AB - NCBI completed the transition of its main genome annotation database from Locuslink to Entrez Gene in Spring 2005. However, to this date few parsers exist for the Entrez Gene annotation file. Owing to the widespread use of Locuslink and the popularity of Perl programming language in bioinformatics, a publicly available high performance Entrez Gene parser in Perl is urgently needed. We present four such parsers that were developed using several parsing approaches (Parse::RecDescent, Parse::Yapp, Perl-byacc and Perl 5 regular expressions) and provide the first in-depth comparison of these sophisticated Perl tools. Our fastest parser processes the entire human Entrez Gene annotation file in under 12 min on one Intel Xeon 2.4 GHz CPU and can be of help to the bioinformatics community during and after the transition from Locuslink to Entrez Gene. PMID- 15879452 TI - A Bayesian regression approach to the inference of regulatory networks from gene expression data. AB - MOTIVATION: There is currently much interest in reverse-engineering regulatory relationships between genes from microarray expression data. We propose a new algorithmic method for inferring such interactions between genes using data from gene knockout experiments. The algorithm we use is the Sparse Bayesian regression algorithm of Tipping and Faul. This method is highly suited to this problem as it does not require the data to be discretized, overcomes the need for an explicit topology search and, most importantly, requires no heuristic thresholding of the discovered connections. RESULTS: Using simulated expression data, we are able to show that this algorithm outperforms a recently published correlation-based approach. Crucially, it does this without the need to set any ad hoc threshold on possible connections. PMID- 15879453 TI - PMUT: a web-based tool for the annotation of pathological mutations on proteins. AB - PMUT allows the fast and accurate prediction (approximately 80% success rate in humans) of the pathological character of single point amino acidic mutations based on the use of neural networks. The program also allows the fast scanning of mutational hot spots, which are obtained by three procedures: (1) alanine scanning, (2) massive mutation and (3) genetically accessible mutations. A graphical interface for Protein Data Bank (PDB) structures, when available, and a database containing hot spot profiles for all non-redundant PDB structures are also accessible from the PMUT server. PMID- 15879454 TI - GMCheck: Bayesian error checking for pedigree genotypes and phenotypes. AB - GMCheck uses graphical modeling to find the posterior probabilities of data errors given genotypes or phenotypes in a specified pedigree structure. PMID- 15879455 TI - Ensemble dependence model for classification and prediction of cancer and normal gene expression data. AB - MOTIVATION: DNA microarray technologies make it possible to simultaneously monitor thousands of genes' expression levels. A topic of great interest is to study the different expression profiles between microarray samples from cancer patients and normal subjects, by classifying them at gene expression levels. Currently, various clustering methods have been proposed in the literature to classify cancer and normal samples based on microarray data, and they are predominantly data-driven approaches. In this paper, we propose an alternative approach, a model-driven approach, which can reveal the relationship between the global gene expression profile and the subject's health status, and thus is promising in predicting the early development of cancer. RESULTS: In this work, we propose an ensemble dependence model, aimed at exploring the group dependence relationship of gene clusters. Under the framework of hypothesis-testing, we employ genes' dependence relationship as a feature to model and classify cancer and normal samples. The proposed classification scheme is applied to several real cancer datasets, including cDNA, Affymetrix microarray and proteomic data. It is noted that the proposed method yields very promising performance. We further investigate the eigenvalue pattern of the proposed method, and we discover different patterns between cancer and normal samples. Moreover, the transition between cancer and normal patterns suggests that the eigenvalue pattern of the proposed models may have potential to predict the early stage of cancer development. In addition, we examine the effects of possible model mismatch on the proposed scheme. PMID- 15879456 TI - Algorithms for alignment of mass spectrometry proteomic data. AB - MOTIVATION: The analysis of biological samples with high-throughput mass spectrometers has increased greatly in recent years. As larger datasets are processed, it is important that the spectra are aligned to ensure that the same protein intensities are correctly identified in each sample. Without such an alignment procedure it is possible to make errors in identifying the signals from peptides with similar molecular weight. Two algorithms are provided that can improve the alignment among samples. One algorithm is designed to work with SELDI data produced from a Ciphergen instrument, and the other can be used with data in a more general format. RESULTS: The two algorithms were applied to samples drawn from a common pool of reference serum. The results indicate substantial improvement in consistently identifying peptide signals in different samples. PMID- 15879457 TI - SDMinP: a program to control the family wise error rate using step-down minP adjusted P-values. AB - SDMinP is an easy-to-use program for fast calculation of empirical and adjusted P values for correlated and uncorrelated hypotheses in multiple testing experiments. It is based on the Free Step-Down Resampling Method for controlling the family wise error rate, and implements a variation of an efficient algorithm, which reduces the originally required re-sampling effort considerably and makes the method computationally feasible. The program is independent of the underlying test statistic and works with provided observed and permutation test statistics. PMID- 15879458 TI - MEPD: a resource for medaka gene expression patterns. AB - The Medaka Expression Pattern Database (MEPD) is a database for gene expression patterns determined by in situ hybridization in the small freshwater fish medaka (Oryzias latipes). Data have been collected from various research groups and MEPD is developing into a central expression pattern depository within the medaka community. Gene expression patterns are described by images and terms of a detailed medaka anatomy ontology of over 4000 terms, which we have developed for this purpose and submitted to Open Biological Ontologies. Sequences have been annotated via BLAST match results and using Gene Ontology terms. These new features will facilitate data analyses using bioinformatics approaches and allow cross-species comparisons of gene expression patterns. Presently, MEPD has 19,757 entries, for 1024 of them the expression pattern has been determined. PMID- 15879459 TI - Pulmonary reexpansion causes xanthine oxidase-induced apoptosis in rat lung. AB - The pathogenesis of reexpansion pulmonary edema is not yet fully understood. We therefore studied its mechanism in a rat model in which the left lung was collapsed by bronchial occlusion for 1 h and then reexpanded and ventilated for an additional 3 h. We then evaluated the production of reactive oxygen species in the lungs using fluorescent imaging and cerium deposition electron microscopic techniques and the incidence of apoptosis using the TdT-mediated dUTP-digoxigenin nick end labeling (TUNEL) method. We found that pulmonary reexpansion induced production of reactive oxygen species and then apoptosis, mainly in endothelial and alveolar type II epithelial cells. Endothelial cells and alveolar type I and II epithelial cells in the reexpanded lung were positive for TUNEL and cleaved caspase-3. DNA fragmentation was also observed in the reexpanded lung. In addition, wet-dry ratios obtained with reexpanded lungs were significantly higher than those obtained with control lungs, indicating increased fluid content. All of these effects were attenuated by pretreating rats with a specific xanthine oxidase inhibitor, sodium (-)-8-(3-methoxy-4-phenylsulfinylphenyl) pyrazolo[1,5 a]-1,3,5-triazine-4(1H)-one. It thus appears that pulmonary reexpansion activates xanthine oxidase in both endothelial and alveolar type II epithelial cells and that the reactive oxygen species produced by the enzyme induce apoptosis among the endothelial and alveolar type I and II epithelial cells that make up the pulmonary water-air barrier, leading to reexpansion pulmonary edema. PMID- 15879460 TI - Pressure-induced leukocyte margination in lung postcapillary venules. AB - Although pressure elevation in lung postcapillary venules increases endothelial P selectin expression, the extent to which P-selectin causes lung leukocyte margination remains controversial. To address this issue, we optically viewed postcapillary venules of the isolated blood-perfused rat lung by real-time fluorescence imaging. To determine leukocyte margination in single postcapillary venules, we quantified the fluorescence of leukocytes labeled in situ with rhodamine 6G (R6G). Although baseline fluorescence was sparse, a 10-min pressure elevation by 10 cmH(2)O markedly increased R6G fluorescence. Both stopping blood flow during pressure elevation and eliminating leukocytes from the perfusion blocked the fluorescence increase, affirming that these fluorescence responses were attributable to pressure-induced leukocyte margination. A P-selectin blocking MAb and the L- and P-selectin blocker fucoidin each inhibited the fluorescence increase, indicating that P-selectin was critical for inducing margination. Time-dependent imaging of blood-borne fluorescent beads revealed reduction of plasma velocity during pressure elevation. After pressure returned to baseline, a similar reduction of plasma velocity, established by manually decreasing the perfusion rate, prolonged margination. Our findings show that in lung postcapillary venules, the decrease in plasma velocity critically determines pressure-induced leukocyte margination. PMID- 15879461 TI - Temporal and spatial distribution of ciliogenesis in the tracheobronchial airways of mice. AB - Little is known about ciliogenesis as it proceeds through the entire airway tree, from the trachea to the terminal bronchioles, especially during the postnatal period. The purpose of this study was to define the spatial and temporal (prenatal and postnatal) pattern of normal cilia development in the mouse. Three airway generations representing the entire airway tree were examined: trachea, lobar bronchi, and terminal bronchiole. Ciliated cells in lung lobe whole mounts were labeled with a fluorescent dye for confocal microscopy, and ciliated cell surface density was measured for each airway generation and age. The same samples were examined by scanning electron microscopy to verify the appearance of ciliated cells among the differentiating epithelium of the airways. Ciliated cells were first detected in the trachea and lobar bronchi at 16 days gestational age (DGA) and in the terminal bronchioles at 18 DGA. Ciliated cell surface density increased with prenatal and postnatal age at all airway levels. However, the ciliated cell surface density of the trachea and lobar bronchi was always greater compared with the terminal bronchiole. In conclusion, the study revealed that in developing tracheobronchial airways of the mouse: 1) Ciliogenesis differs temporally and spatially by airway generation; 2) Ciliated cell surface density increases with age in all airway generations, but density decreases in a proximal to distal direction; and 3) A significant portion of ciliogenesis continues after birth. This study provides a healthy basis for investigations of neonatal pulmonary disease or pollutant toxicity affecting cilia and its functions. PMID- 15879462 TI - Bisphenol-A induces cell cycle delay and alters centrosome and spindle microtubular organization in oocytes during meiosis. AB - Bisphenol-A (BPA) is a widely used environmental estrogen-like chemical that has a weak estrogenic activity. This study aimed to test the potential inhibitory effects of BPA on meiotic cell cycle progression, centrosomes and spindle integrity in mouse cumulus-oocyte complexes (COCs). They were exposed to BPA (10 30 microM; 2.3-6.8 ppm) during meiosis-I and the formation of metaphase-II (M-II) spindle. Exposure to BPA during meiosis-I caused a dose-dependent retardation/inhibition of cell cycle progression; 74 and 61% of cells reached metaphase-I (M-I) in the presence of 10 and 30 microM BPA, respectively, (81% in controls, P<0.001). A more striking delay was noted when oocytes were exposed to BPA during the formation of M-II spindle, i.e. 61 and 41% of cells (94% in controls, P<0.001) reached M-II while the remaining cells remained at M-I. Depending on dose, both (i) loosening and elongation of meiotic spindles and (ii) compaction and dispersion of pericentriolar material (PCM) were noted in all samples, all of which resulted in a series of spindle abnormalities. Interestingly, no chromosome was detected in the first polar body after the 10 and 30 microM BPA treatments. When the cells were freed from BPA exposure at 10 and 30 microM, 70 and 61%, of the cells succeeded in reaching M-II (93% in controls, P<0.001), respectively. In conclusion, one mode of action of BPA is a moderately severe yet reversible delay in the meiotic cell cycle, possibly by a mechanism that degrades centrosomal proteins and thus perturbs the spindle microtubule organization and chromosome segregation. PMID- 15879463 TI - Association of SHBG gene polymorphism with menarche. AB - The age of menarche may be subject to hereditary influences but the specific determinants are unknown. Our aim was to investigate the possible association of a functional (TAAAA)n polymorphism in the promoter of the sex hormone-binding globulin (SHBG) gene with the timing of menarche. This polymorphism has been associated with polycystic ovary syndrome (PCOS) and is considered to contribute to SHBG levels. We studied 130 healthy normal-weight adolescent females from a closed community in North-Western Greece. Information on menarche was obtained through interviews. The BMI was recorded. Genomic DNA was isolated from peripheral blood leukocytes for genotyping the TAAAA repeat region. We subdivided our subjects into two groups based on median age of menarche: those with menarche <13 years and those with menarche > or =13 years. Genotype analysis revealed six (TAAAA)n alleles containing 5-10 TAAAA repeats. The distribution of alleles was different in the two groups. Girls with late menarche had more frequently longer TAAAA alleles (>8 repeats), while girls with early menarche had shorter alleles at a greater frequency (P=0.048). The major contribution to early menarche was by the 6 TAAAA repeat allele. Furthermore, carriers of the longer allele genotypes had later menarche (13.24+/-1.15 years) than those with shorter allele genotypes (12.67+/-1.15, P=0.018). These findings provide evidence for a genetic contribution of SHBG gene to the age of menarche. PMID- 15879464 TI - The function of the SNP in the MMP1 and MMP3 promoter in susceptibility to endometriosis in China. AB - Matrix metalloproteinases (MMPs) may contribute to the development of endometriosis. Genetic variations in several MMP promoters may influence the transcription and expression of MMPs. The purpose of the present study was to assess how gene polymorphisms in the MMP1 and MMP3 promoters affect the risk of development of endometriosis. We genotyped 100 women with endometriosis and 150 control subjects in North China. There was a significant difference in frequency of the MMP1 genotype between cases and controls (P=0.03). The 2G homozygote in endometriosis and controls was significantly different (P=0.02). The frequency of the 2G allele among affected women (79%) was significantly higher than among the healthy controls (66.9%; P=0.003). However, the overall genotype and allelotype distribution of the MMP3 single nucleotide polymorphism (SNP) in patients was not different from that of controls (P> or =0.05). MMP1 and MMP3 polymorphisms were in linkage disequilibrium in cases and controls (D'=0.47; P=0.00). The haplotype frequency distribution derived from these two polymorphisms was significantly different between cases and controls (P=0.00). The haplotype analysis suggested an implication of both MMP1 and MMP3 polymorphisms in the susceptibility to endometriosis. We conclude that the MMP1 promoter SNP and MMP 2G/6A haplotype may modify susceptibility to endometriosis, but that the MMP3 promoter SNP is unlikely to be associated with endometriosis in the population of North China. PMID- 15879465 TI - Estrogen-induced changes in IGF-I, Myb family and MAP kinase pathway genes in human uterine leiomyoma and normal uterine smooth muscle cell lines. AB - Many studies have implicated numerous hormones, growth factors, cytokines and other signal transduction molecules in the pathogenesis of uterine leiomyoma. Estrogen and estrogen-related genes are thought to play a key role in the growth of uterine leiomyomas, but the molecular mechanisms are unclear. In an attempt to investigate various pathways that might be involved in estrogen-regulated uterine leiomyoma growth as well as to identify any novel effector genes, microarray studies comparing estrogen-treated uterine leiomyoma cells (UtLM) and normal myometrial cells to untreated cells were performed. Several genes were differentially expressed in estrogen treated UtLM cells, including insulin-like growth factor-I (IGF-I) and others potentially involved in the IGF-I signalling pathway, specifically genes for A-myb, a transcription factor which promotes cell cycle progression and for MKP-1, a dual specificity phosphatase that dephosphorylates mitogen-activated protein kinase. IGF-I and A-myb were up regulated in estrogen-treated cells while MKP-1 was down-regulated. Two other cell cycle promoting genes, c-fos and myc, were also down-regulated in estrogen treated UtLM cells. These genes are typically up-regulated in response to estrogen in some cells, notably breast epithelial cells, yet consistently have lower expression levels in uterine leiomyoma tissue when compared to autologous myometrium. Our results demonstrate some novel genes that may play a role in the growth of uterine leiomyoma, strengthen the case for involvement of the IGF-I pathway in the response of UtLM to estrogen and corroborate evidence that uterine smooth muscle cells respond to estrogen with a different gene expression pattern than that seen in epithelial cells. PMID- 15879466 TI - DAZL expression in human oocytes, preimplantation embryos and embryonic stem cells. AB - In humans, the Deleted in Azoospermia Like (DAZL) gene is believed to function in the development of primordial germ cells and in germ cell differentiation and maturation because the expression of DAZL is only found in the germ and non-germ lineage of the reproductive system and in embryonic stem (ES) cells. The present study examined the presence of DAZL transcripts in the last stages of oocyte maturation, in ES cells, and throughout the preimplantation development; the link between gametes and ES cells. The finding of DAZL transcripts in the last stages of oogenesis and during the first two cell cycles of the preimplantation development was expected, because DAZL is a germ cell marker and the transcripts present at that time are generally encoded by the maternal genome. During the third cell cycle, DAZL showed a variable expression pattern, which may point to the maternal to embryonic transition. After the third cell cycle, transcripts were again consistently detected, suggesting embryonic DAZL transcription. In blastocysts, DAZL transcripts were only detected in those of good quality and this as well in the inner cell mass (ICM) as in the trophectoderm (TE). The presence of DAZL transcripts in the ICM and in ES cells was not surprising since both can lead to the formation of germ cells, but TE cells cannot. The quality related expression of DAZL in blastocysts, and especially its trophectodermal expression, might imply other functions for DAZL beyond germ cell development. PMID- 15879467 TI - Main phase transitions in supported lipid single-bilayer. AB - We have studied the phase transitions of a phospholipidic single-bilayer supported on a mica substrate by real-time temperature-controlled atomic force microscopy. We show the existence of two phase transitions in this bilayer that we attribute to two gel (L(beta))/fluid (L(alpha)) transitions, corresponding to the independent melting of each leaflet of the bilayer. The ratio of each phase with temperature and the large broadening of the transitions' widths have been interpreted through a basic thermodynamic framework in which the surface tension varies during the transitions. The experimental data can be fit with such a model using known thermodynamic parameters. PMID- 15879468 TI - Testing two predictions for fracture load using computer models of trabecular bone. AB - Aging induces several types of architectural changes in trabecular bone including thinning, increased levels of anisotropy, and perforation. It has been determined, on the basis of analysis of mathematical models, that reduction in fracture load caused by perforation is significantly higher than those due to equivalent levels of thinning or anisotropy. The analysis has also provided an expression which relates the fractional reduction of strength tau to the fraction of elements nu that have been removed from a network. Further, it was proposed that the ratio Gamma of the elastic constant of a sample and its linear response at resonance can be used as a surrogate for tau. Experimental validation of these predictions requires following architectural changes in a given sample of trabecular bone; techniques to study such changes using microcomputed tomography are only beginning to be available. In the present study, we use anatomically accurate computer models constructed from digitized images of bone samples for the purpose. Images of healthy bone are subjected to successive levels of synthetic degradation via surface erosion. Computer models constructed from these images are used to calculate their fracture load and other mechanical properties. Results from these computations are shown to be consistent with predictions derived from the analysis of mathematical models. Although the form of tau(nu) is known, parameters in the expression are expected to be sample-specific, and hence nu is not a reliable predictor of strength. We provide an example to demonstrate this. In contrast, analysis of model networks shows that the linear part of tau(Gamma) depends only on the structure of trabecular bone. Computations on models constructed from samples of iliac crest trabecular bone are shown to be in agreement with this assertion. Since Gamma can be computed from a vibrational assessment of bone, we argue that the latter can be used to introduce new surrogates for bone strength and hence diagnostic tools for osteoporosis. PMID- 15879469 TI - Local mobility in lipid domains of supported bilayers characterized by atomic force microscopy and fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) is used to examine mobility of labeled probes at specific sites in supported bilayers consisting of 1,2 dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid domains in 1,2-dioleoyl-sn glycero-3-phosphocholine (DOPC). Those sites are mapped beforehand with simultaneous atomic force microscopy and submicron confocal fluorescence imaging, allowing characterization of probe partitioning between gel DPPC and disordered liquid DOPC domains with corresponding topography of domain structure. We thus examine the relative partitioning and mobility in gel and disordered liquid phases for headgroup- and tailgroup-labeled GM1 ganglioside probes and for headgroup- and tailgroup-labeled phospholipid probes. For the GM1 probes, large differences in mobility between fluid and gel domains are observed; whereas unexpected mobility is observed in submicron gel domains for the phospholipid probes. We attribute the latter to domain heterogeneities that could be induced by the probe. Furthermore, fits to the FCS data for the phospholipid probes in the DOPC fluid phase require two components (fast and slow). Although proximity to the glass substrate may be a factor, local distortion of the probe by the fluorophore could also be important. Overall, we observe nonideal aspects of phospholipid probe mobility and partitioning that may not be restricted to supported bilayers. PMID- 15879470 TI - Sacrificial bonds in polymer brushes from rat tail tendon functioning as nanoscale velcro. AB - Polymers play an important role in many biological systems, so a fundamental understanding of their cross-links is crucial not only for the development of medicines but also for the development of biomimetic materials. The biomechanical movements of all mammals are aided by tendon fibrils. The self-organization and biomechanical functions of tendon fibrils are determined by the properties of the cross-links between their individual molecules and the interactions among the cross-links. The cross-links of collagen and proteoglycan molecules are particularly important in tendons and, perhaps, bone. To probe cross-links between tendon molecules, we used the cantilever tip of an atomic force microscope in a pulling setup. Applying higher forces to rat tail tendon molecules with the tip led to a local disruption of the highly organized shell of tendon fibrils and to the formation or an increase of a polymer brush of molecules sticking out of the surface. The cross-linking between these molecules was influenced by divalent Ca2+ ions. Furthermore, the molecules of the polymer brush seemed to bind back to the fibrils in several minutes. We propose that sacrificial bonds significantly influence the tendon fibrils' self-organization and self-healing and therefore contribute to toughness and strength. PMID- 15879471 TI - Effect of microvillus deformability on leukocyte adhesion explored using adhesive dynamics simulations. AB - Leukocyte rolling on the endothelium via selectin molecules is an important step in the adhesion cascade, which allows leukocytes in the bloodstream to reach sites of infection. We improve upon Adhesive Dynamics simulations by incorporating deformable microvilli on which adhesion molecules are clustered. As determined in micropipette experiments, microvilli deform like an elastic spring at small forces and a combination of yield and viscous dissipation at high forces. First, we create a modified version of the state diagram for adhesion which includes microvillus deformation, and find four adhesion states-firmly bound; landing; rolling; and no-adhesion. Then, we simulate the effects of receptor clustering on the tips of microvilli, number of adhesion molecules on the cell, and the spring constant of the bonds, within the context of deformable microvilli. We also explore how the microvillus rheology itself controls the dynamics of adhesion. A minimum in rolling velocity occurs at an intermediate value of the microvillus membrane viscosity, remarkably close to the reported physiological value, suggesting that the mechanics of microvilli have evolved ideally for rolling and adhesion of leukocytes. We find that a larger degree of association between the membrane and cytoskeleton leads to slower rolling, and stiffer microvilli result in faster rolling. Decreasing the overall deformability of the microvilli greatly reduces a simulated cell's ability to roll. A comparison to experimental results of in vitro cell rolling agrees with the simulation at low shear rates. Furthermore, simulated rolling trajectories of cells with deformable microvilli display periods of rolling interdispersed with pauses, consistent with that seen in experiments where microvilli were observed to stretch. PMID- 15879472 TI - Large-scale quantitative analysis of sources of variation in the actin polymerization-based movement of Listeria monocytogenes. AB - During the actin polymerization-based movement of Listeria monocytogenes, individual bacteria are rapidly propelled through the host cell cytoplasm by the growth of a filamentous actin tail. The rate of propulsion varies significantly among individuals and over time. To study this variation, we used a high throughput tracking technique to record the movement of a large number (approximately 7900) of bacteria in Xenopus frog egg extract. Most bacteria (70%) appeared to maintain an individual characteristic speed over several minutes, suggesting that the major source of variation in average speed is intrinsic to the bacterium. Thirty percent of bacteria had significant changes in speed over time spans of a few minutes, including 17% that appeared to collide with obstacles and 13% that moved with a significant periodic component. For the latter, the peak frequency was proportional to speed, suggesting a mechanism with a fixed spatial scale of approximately 0.6 bacterial length. Near the rear of the bacterium, temporal fluctuations in actin density were positively correlated with fluctuations in speed, whereas near the front the correlation was negative. A comparison of the performance of linear models that predict motion given actin density suggests that the mechanism has a history of 5-10 s, and that fluctuations in actin density near the front of the bacteria contain more predictive information than the rear. Our results are consistent with physical models where bacterial speed is governed by the rate of dissociation of bonds between the bacterial surface and the actin tail, and individual variation is determined by long-lived intrinsic variability in bacterial surface properties. PMID- 15879473 TI - The phot LOV2 domain and its interaction with LOV1. AB - Phot proteins are homologs of the blue-light receptor phototropin. We report a comparative study of the photocycles of the isolated, light-sensitive domains LOV1 and LOV2 from Chlamydomonas reinhardtii phot protein, as well as the construct LOV1/2 containing both domains. Transient absorption measurements revealed a short lifetime of the LOV2-wt triplet state (500 ns), but a long lifetime (287 micros) of the triplet in the mutant LOV2-C250S, in which the reactive cysteine is replaced by serine. For LOV1, in comparison, corresponding numbers of 800 ns and 4 micros for the two conformers in LOV1-wt, and 27 micros for LOV1-C57S have been reported. The triplet decay kinetics in the mixed domains LOV1/2-wt, LOV1/2-C57S, and LOV1/2-C250S can be analyzed as the superposition of the behavior of the corresponding single domains. The situation is different for the slow, thermal reaction of the photoadduct back to the dark form. Whereas the individual domains LOV1 and LOV2 show two decay components, the double domains LOV1/2-C57S and LOV1/2-C250S both show only a single component. The interaction of the two domains does therefore not manifest itself during the lifetime of the triplet states, but changes the decay behavior of the adduct states. PMID- 15879474 TI - The physics of filopodial protrusion. AB - Filopodium, a spike-like actin protrusion at the leading edge of migrating cells, functions as a sensor of the local environment and has a mechanical role in protrusion. We use modeling to examine mechanics and spatial-temporal dynamics of filopodia. We find that >10 actin filaments have to be bundled to overcome the membrane resistance and that the filopodial length is limited by buckling for 10 30 filaments and by G-actin diffusion for >30 filaments. There is an optimal number of bundled filaments, approximately 30, at which the filopodial length can reach a few microns. The model explains characteristic interfilopodial distance of a few microns as a balance of initiation, lateral drift, and merging of the filopodia. The theory suggests that F-actin barbed ends have to be focused and protected from capping (the capping rate has to decrease one order of magnitude) once every hundred seconds per micron of the leading edge to initiate the observed number of filopodia. The model generates testable predictions about how filopodial length, rate of growth, and interfilopodial distance should depend on the number of bundled filaments, membrane resistance, lamellipodial protrusion rate, and G-actin diffusion coefficient. PMID- 15879475 TI - Cholesterol-enriched lipid domains can be visualized by di-4-ANEPPDHQ with linear and nonlinear optics. AB - We present a membrane-staining dye, di-4-ANEPPDHQ, which differentiates liquid ordered phases from liquid-disordered phases coexisting in model membranes under both linear and nonlinear microscopies. The dye's fluorescence emission spectrum is blue-shifted 60 nm in liquid-ordered phases compared with liquid-disordered phases, and shows strong second harmonic generation in the liquid-disordered phase compared with the liquid-ordered phase. The ease of staining and the ability of this single dye to detect both phases, should lead to broad applications in biophysical studies of lipid domains in model membranes and cells. PMID- 15879476 TI - Structural transformations in the dynamics of Michaelis complex formation in lactate dehydrogenase. AB - The dynamical nature of the binding of a substrate surrogate to lactate dehydrogenase is examined on the nanoseconds to milliseconds timescale by laser induced temperature-jump relaxation spectroscopy. Fluorescence emission of the nicotinamide group of bound NADH is used to define the pathway and kinetics of substrate binding. Assignment of specific kinetic states and elucidation of their structures are accomplished using isotope edited infrared absorption spectroscopy. Such studies are poised to yield a detailed picture of the coupling of protein dynamics to function. PMID- 15879477 TI - Probing the local dynamics of nucleotide-binding pocket coupled to the global dynamics: myosin versus kinesin. AB - Based on the elastic network model, we develop a new analysis for protein complexes, which probes the local dynamics of a subsystem that is elastically coupled to a fluctuating environment. This method is applied to a comparative dynamical analysis of the nucleotide-binding pocket of two motor proteins-myosins and kinesins. In myosins, the observed structural changes in the nucleotide pocket from the transition state to the rigorlike state are dominated by the lowest normal mode that involves significant movements in both switch I and switch II; in kinesins, the measured conformational changes in the nucleotide pocket are also dominated by the lowest mode, which, however, only involves large movement in switch I. We then compute the global structural changes induced by the nucleotide-pocket deformations as described by the dominant pocket-mode, which yield encouraging results: in myosins, multiple hinge motions involving the opening/closing of the cleft between the upper and lower 50 -kDa subdomains and the swinging movement of the converter are induced, which are dominated by precisely the same global mode that has been recently identified by us as important to the dynamical correlations among the nucleotide-pocket, the actin binding site, and the converter; in kinesins, the induced global conformational changes are well described by a highly collective global mode which hints for a dynamical pathway spanning from the nucleotide-pocket to the neck-linker via the H6 helix. PMID- 15879478 TI - Stoichiometry and substrate affinity of the mannitol transporter, EnzymeIImtl, from Escherichia coli. AB - Uptake and consecutive phosphorylation of mannitol in Escherichia coli is catalyzed by the mannitol permease EnzymeIImtl. The substrate is bound at an extracellular-oriented binding site, translocated to an inward-facing site, from where it is phosphorylated, and subsequently released into the cell. Previous studies have shown the presence of both a high- and a low-affinity binding site with K(D)-values in the nano- and micromolar range, respectively. However, reported K(D)-values in literature are highly variable, which casts doubts about the reliability of the measurements and data analysis. Using an optimized binding measurement system, we investigated the discrepancies reported in literature, regarding both the variability in K(D)-values and the binding stoichiometry. By comparing the binding capacity obtained with flow dialysis with different methods to determine the protein concentration (UV-protein absorption, Bradford protein detection, and a LDH-linked protein assay to quantify the number of phosphorylation sites), we proved the existence of only one mannitol binding site per dimeric species of unphosphorylated EnzymeIImtl. Furthermore, the affinity of EnzymeIImtl for mannitol appeared to be dependent on the protein concentration and seemed to reflect the presence of an endogenous ligand. The dependency could be simulated assuming that >50% of the binding sites were occupied with a ligand that shows an affinity for EnzymeIImtl in the same range as mannitol. PMID- 15879479 TI - Cortical actomyosin breakage triggers shape oscillations in cells and cell fragments. AB - Cell shape and movements rely on complex biochemical pathways that regulate actin, microtubules, and substrate adhesions. Some of these pathways act through altering the cortex contractility. Here we examined cellular systems where contractility is enhanced by disassembly of the microtubules. We found that adherent cells, when detached from their substrate, developed a membrane bulge devoid of detectable actin and myosin. A constriction ring at the base of the bulge oscillated from one side of the cell to the other. The movement was accompanied by sequential redistribution of actin and myosin to the membrane. We observed this oscillatory behavior also in cell fragments of various sizes, providing a simplified, nucleus-free system for biophysical studies. Our observations suggest a mechanism based on active gel dynamics and inspired by symmetry breaking of actin gels growing around beads. The proposed mechanism for breakage of the actomyosin cortex may be used for cell polarization. PMID- 15879480 TI - Mechanisms of enhanced shock-induced arrhythmogenesis in the rabbit heart with healed myocardial infarction. AB - Shock-induced vulnerability and defibrillation have been mostly studied in structurally normal hearts. However, defibrillation therapy is normally applied to patients with diseased hearts, frequently those with prior myocardial infarction (MI). Shock-induced vulnerability and defibrillation have not been well studied under this condition. We sought to examine the mechanisms of shock induced arrhythmogenesis and arrhythmia maintenance in a rabbit model of healed MI (4 wk or more postinfarction). Ligation of the lateral division or posterolateral division of the left coronary artery at a level of 40-70% from the apex was performed 53 +/- 21 days before acute experiments. Shock-induced vulnerability was assessed in infarcted (n = 8) and structurally normal (n = 8) hearts by delivering internal monophasic shocks at different shock strengths and delivery phases. Electrical activities from the anterior epicardium during shock application and during shock-induced arrhythmias were optically recorded and quantitatively analyzed. Ligation resulted in a transmural left ventricular free wall infarction mainly located at the apical region with a consistent endocardial border zone (BZ) as confirmed by histological studies. There were significant increases in the incidence, severity, and duration of shock-induced arrhythmias in the infarcted hearts versus controls due to 1) postshock break-excitation wavefronts that frequently originated near the infarction BZ and 2) the existence of an infarction BZ that created an anatomic reentry pathway and facilitated arrhythmia maintenance. In conclusion, the infarction BZ contributes to both increased shock-induced arrhythmogenesis and arrhythmia maintenance in the rabbit model of healed MI. PMID- 15879481 TI - Reduced functional expression of K(+) channels in vascular smooth muscle cells from rats made hypertensive with N{omega}-nitro-L-arginine. AB - Smooth muscle membrane potential is determined, in part, by K(+) channels. In the companion paper to this article, we demonstrated that superior mesenteric arteries from rats made hypertensive with N(omega)-nitro-l-arginine (l-NNA) are depolarized and express less K(+) channel protein compared with those from normotensive rats. In the present study, we used patch-clamp techniques to test the hypothesis that l-NNA-induced hypertension reduces the functional expression of K(+) channels in smooth muscle. In whole cell experiments using a Ca(2+)-free pipette solution, current at 0 mV, largely due to voltage-dependent K(+) (K(V)) channels, was reduced approximately 60% by hypertension (2.7 +/- 0.4 vs. 1.1 +/- 0.2 pA/pF). Current at +100 mV with 300 nM free Ca(2+), largely due to large conductance Ca(2+)-activated K(+) (BK(Ca)) channels, was reduced approximately 40% by hypertension (181 +/- 24 vs. 101 +/- 28 pA/pF). Current blocked by 3 mM 4 aminopyridine, an inhibitor of many K(V) channel types, was reduced approximately 50% by hypertension (1.0 +/- 0.4 vs. 0.5 +/- 0.2 pA/pF). Current blocked by 1 mM tetraethylammonium, an inhibitor of BK(Ca) channels, was reduced approximately 40% by hypertension (86 +/- 14 vs. 53 +/- 19 pA/pF). Differences in BK(Ca) current magnitude are not attributable to changes in single-channel conductance or Ca(2+)/voltage sensitivity. The data support the hypothesis that l-NNA-induced hypertension reduces K(+) current in vascular smooth muscle. Reduced molecular and functional expression of K(+) channels may partly explain the depolarization and augmented contractile sensitivity of smooth muscle from l-NNA-treated rats. PMID- 15879482 TI - Calcitonin gene-related peptide and substance P contribute to reduced blood pressure in sympathectomized rats. AB - CGRP and substance P (SP) are produced in dorsal root ganglia (DRG) sensory neurons and modulate vascular tone. Sympathetic and sensory nerves compete for NGF, a potent stimulator of CGRP and SP, and it has been suggested that sympathetic hyperinnervation in spontaneously hypertensive rats may reduce the availability of NGF to sensory nerves, thus reducing CGRP and SP. The purpose of this study was to determine whether destruction of peripheral sympathetic nerves in normal rats would increase the availability of NGF for sensory neurons and enhance expression of CGRP and SP. Sympathectomy was produced in rats by guanethidine sulfate administration. Control rats received saline. Sympathectomized rats displayed reductions in blood pressure (BP) and atria norepinephrine levels, whereas NGF levels in the DRG, spleen, and ventricles were increased. Sympathectomy also enhanced CGRP and SP mRNA and peptide content in DRG. Administration of CGRP and SP receptor antagonists increased the BP in sympathectomized rats but not in the controls. Thus sympathectomy enhances sensory neuron CGRP and SP expression that contributes to the BP reduction. PMID- 15879483 TI - Measurement of total pulmonary arterial compliance using invasive pressure monitoring and MR flow quantification during MR-guided cardiac catheterization. AB - Pulmonary hypertensive disease is assessed by quantification of pulmonary vascular resistance. Pulmonary total arterial compliance is also an indicator of pulmonary hypertensive disease. However, because of difficulties in measuring compliance, it is rarely used. We describe a method of measuring pulmonary arterial compliance utilizing magnetic resonance (MR) flow data and invasive pressure measurements. Seventeen patients with suspected pulmonary hypertension or congenital heart disease requiring preoperative assessment underwent MR-guided cardiac catheterization. Invasive manometry was used to measure pulmonary arterial pressure, and phase-contrast MR was used to measure flow at baseline and at 20 ppm nitric oxide (NO). Total arterial compliance was calculated using the pulse pressure method (parameter optimization of the 2-element windkessel model) and the ratio of stroke volume to pulse pressure. There was good agreement between the two estimates of compliance (r = 0.98, P < 0.001). However, there was a systematic bias between the ratio of stroke volume to pulse pressure and the pulse pressure method (bias = 61%, upper level of agreement = 84%, lower level of agreement = 38%). In response to 20 ppm NO, there was a statistically significant fall in resistance, systolic pressure, and pulse pressure. In seven patients, total arterial compliance increased >10% in response to 20 ppm NO. As a population, the increase did not reach statistical significance. There was an inverse relation between compliance and resistance (r = 0.89, P < 0.001) and between compliance and mean pulmonary arterial pressure (r = 0.72, P < 0.001). We have demonstrated the feasibility of quantifying total arterial compliance using an MR method. PMID- 15879484 TI - Concomitant antagonism of endothelial and vascular smooth muscle cell ETB receptors for endothelin induces hypertension in the hamster. AB - In the vascular system, endothelin (ET) type B (ET(B)) receptors for ET-1 are located on endothelial and on venous and arterial smooth muscle cells. In the present study, we investigated the hemodynamic effects of chronic ET(B) receptor blockade at low and high doses in the Syrian Golden hamster. After 16 days of gavage with A-192621 (0.5 or 30 mg.kg(-1).day(-1)), a selective ET(B) receptor antagonist, hamsters were anesthetized with a mixture of ketamine and xylazine (87 and 13 mg/kg im, respectively), and basal mean arterial blood pressure (MAP) and pressor responses to exogenous ET-1 were evaluated. The lower dose of A 192621 (0.5 mg.kg(-1).day(-1)) did not modify basal MAP, whereas the higher dose (30 mg.kg(-1).day(-1)) increased MAP and plasma ET levels. Radio-telemetry recordings confirmed the increase in MAP induced by the higher dose of A-192621 in conscious hamsters. On the other hand, although the lower dose of A-192621 was devoid of intrinsic pressor effects, it markedly reduced the transient hypotensive phase induced by intravenously injected IRL-1620, a selective ET(B) receptor agonist. Finally, A-192621 (0.5 mg.kg(-1).day(-1)) alone or A-192621 (30 mg.kg(-1).day(-1)) + atrasentan (6 mg.kg(-1).day(-1)), a selective ET(A) receptor antagonist, potentiated the pressor response to exogenous ET-1. Our results suggest that, in the hamster, ET(B) receptors on vascular smooth muscle cells are importantly involved in the clearance of endogenous ET-1, whereas the same receptor type on the endothelium is solely involved in the vasodilatory responses to the pressor peptide. Blockade of endothelial and vascular smooth muscle cell ET(B) receptors triggers a marked potentiation of ET(A)-dependent increases in systemic resistance. PMID- 15879485 TI - Endothelial microparticles affect angiogenesis in vitro: role of oxidative stress. AB - Endothelium-derived microparticles have recently been described as a new marker of endothelial cell dysfunction. Increased levels of circulating microparticles have been documented in inflammatory disorders, diabetes mellitus, and many cardiovascular diseases. Perturbations of angiogenesis play an important role in the pathogenesis of these disorders. We demonstrated previously that isolated endothelial microparticles (EMPs) impair endothelial function in vitro, diminishing acetylcholine-induced vasorelaxation and nitric oxide production by rat aortic rings and simultaneously increasing superoxide production. Herein, using the Matrigel assay of angiogenesis in vitro and a topological analysis of the capillary-like network by human umbilical vein endothelial cells (HUVECs), we investigated the effects of EMPs on formation of the vascular network. All parameters of angiogenesis were affected by treatment for 48 h with isolated EMPs in a concentration of 10(5) but not 10(3) or 10(4) EMPs/ml. The effects included decreases in total capillary length (24%), number of meshes (45%), and branching points (36%) and an increase in mesh area (38%). The positional and topological order indicated that EMPs affect angiogenic parameters uniformly over the capillary network. Treatment with the cell-permeable SOD mimetic Mn(III)tetrakis(4-benzoic acid) porphyrin chloride (Mn-TBAP) partially or completely restored all parameters of angiogenesis affected by EMPs. EMPs reduced cell proliferation rate and increased apoptosis rate in time- and dose-dependent manners, and this phenomenon was also prevented by Mn-TBAP treatment. Our data demonstrate that EMPs have considerable impact on angiogenesis in vitro and may be an important contributor to the pathogenesis of diseases that are accompanied by impaired angiogenesis. PMID- 15879486 TI - Acetylcholine causes endothelium-dependent contraction of mouse arteries. AB - The goal of this study was to determine whether acetylcholine evokes endothelium dependent contraction in mouse arteries and to define the mechanisms involved in regulating this response. Arterial rings isolated from wild-type (WT) and endothelial nitric oxide (NO) synthase knockout (eNOS(-/-)) mice were suspended for isometric tension recording. In abdominal aorta from WT mice contracted with phenylephrine, acetylcholine caused a relaxation that reversed at the concentration of 0.3-3 microM. After inhibition of NO synthase [with N(omega) nitro-l-arginine methyl ester (l-NAME), 1 mM], acetylcholine (0.1-10 microM) caused contraction under basal conditions or during constriction to phenylephrine, which was abolished by endothelial denudation. This contraction was inhibited by the cyclooxygenase inhibitor indomethacin (1 muM) or by a thromboxane A(2) (TxA(2)) and/or prostaglandin H(2) receptor antagonist SQ-29548 (1 microM) and was associated with endothelium-dependent generation of the TxA(2) metabolite TxB(2.) Also, SQ-29548 (1 microM) abolished the reversal in relaxation evoked by 0.3-3 microM acetylcholine and subsequently enhanced the relaxation to the agonist. The magnitude of the endothelium-dependent contraction to acetylcholine (0.1-10 microM) was similar in aortas from WT mice treated in vitro with l-NAME and from eNOS(-/-) mice. In addition, we found that acetylcholine (10 microM) also caused endothelium-dependent contraction in carotid and femoral arteries of eNOS(-/-) mice. These results suggest that acetylcholine initiates two competing responses in mouse arteries: endothelium-dependent relaxation mediated predominantly by NO and endothelium-dependent contraction mediated most likely by TxA(2). PMID- 15879487 TI - Transcriptional basis for exercise limitation in male eNOS-knockout mice with age: heart failure and the fetal phenotype. AB - Endothelium-derived nitric oxide (NO) is pivotal in regulating mitochondrial O(2) consumption (Vo(2)) and glucose uptake in mice. The aim of this study was to investigate the mechanism of age- and genotype-related exercise limitation in male endothelial NO synthase (eNOS)-knockout (KO, n = 16) and wild-type (WT, n = 19) mice. Treadmill testing was performed at 12, 14, 16, 18, and 21 mo of age. Vo(2), CO(2) production, respiratory exchange ratio, and maximal running distance were determined during treadmill running. There were good linear correlations for increase of speed with increase of Vo(2). The difference between KO and WT mice was not significant at 12 mo but was significant at 18 mo. Linear regression showed that KO mice consumed more O(2) at the same absolute and relative workloads, suggesting that Vo(2) was not inhibited by NO in KO mice. KO mice performed 30-50% less work than WT mice at each age (work = vertical distance x weight). In contrast to WT mice, the work performed by KO mice significantly decreased from 17 +/- 1.4 m.kg at 12 mo to 9.4 +/- 1.7 m.kg at 21 mo. Running distance was significantly decreased from 334 +/- 27 m at 12 mo to 178 +/- 38 m at 21 mo, and maximal Vo(2), CO(2) production, and respiratory exchange ratio per work unit were significantly higher in KO than in WT mice. Gene arrays showed evidence of a fetal phenotype in KO mice at 21 mo. In conclusion, age- and genotype-related exercise limitations in maximal work performed and maximal running distance in male eNOS-KO mice indicated that fetal phenotype and age were related to onset of heart failure. PMID- 15879488 TI - Acute exercise increases the ventricular arrhythmia threshold via the intrinsic adenosine receptor system in conscious hypertensive rats. AB - Coronary artery occlusion-induced tachyarrhythmias that culminate in ventricular fibrillation are the leading cause of death in developed countries. The intrinsic adenosine receptor system protects the heart from an ischemic insult. Thus the increased functional demands made on the heart during exercise may produce protective adaptations mediated by endogenous adenosine. Therefore, we tested the hypothesis that a single bout of dynamic exercise increases the ventricular arrhythmia threshold (VAT) induced by coronary artery occlusion in conscious hypertensive rats via the intrinsic adenosine receptor system. To test this hypothesis, we recorded the VAT before and on an alternate day after a single bout of dynamic treadmill exercise (12 m/min, 10% grade for 40 min). A single bout of dynamic exercise significantly reduced postexercise arterial pressure (Delta-24 +/- 4 mmHg) and increased VAT (Delta+1.95 +/- 0.31 min). Adenosine receptor blockade with the nonselective adenosine receptor antagonists theophylline or aminophylline (10 mg/kg) attenuated the cardioprotective effects of a single bout of dynamic exercise. Results suggest that strategies that increase myocardial ATP requirements leading to adenosine production provide protection against coronary artery occlusion. PMID- 15879489 TI - Transmural sheet strains in the lateral wall of the ovine left ventricle. AB - In an attempt to provide a better understanding of our finding that regions with contracting left ventricular myofibers need not develop a significant transmural systolic wall thickening gradient, the analytic approach of Costa et al. was applied to the four-dimensional dynamic data obtained 1 and 8 wk after surgical implantation of transmural radiopaque beads in the lateral equatorial left ventricular wall in seven ovine hearts. Quantitative histology of tissue blocks demonstrated that fiber angles varied linearly across the wall in this region from -37 degrees in the subepicardium to +18 degrees in the subendocardium. Sheet angles exhibited a pleated-sheet behavior, alternating sign from subepicardium to subendocardium. From end diastole (reference configuration) to end systole (deformed configuration), fiber strain was uniformly negative, sheet extension and sheet thickening were uniformly positive, and sheet-normal shear contributed to wall thickening at all wall depths. Subepicardial radial wall thickening increased significantly from week 1 to week 8, with significant increases in the contributions from subepicardial sheet extension and sheet-normal shear. At 1 and 8 wk, the contribution of sheet-normal shear to wall thickening was substantial at all transmural depths; the contribution of sheet extension to wall thickening was greatest in the subepicardium and least in the subendocardium, and the contribution of sheet thickening to wall thickening was greatest in the subendocardium and least in the subepicardium. A mechanistic model is proposed that provides a working hypothesis that a selective decrease in subepicardial intercellular matrix stiffness is responsible for elimination of the transmural wall thickening gradient 1-8 wk after marker implantation surgery. PMID- 15879490 TI - Effects of cytokine treatment on angiotensin II type 1A receptor transcription and splicing in rat cardiac fibroblasts. AB - Angiotensin II (ANG II) plays important roles in cardiac extracellular matrix remodeling via its type 1A (AT(1A)) receptor. The cytokines tumor necrosis factor alpha and interleukin-1beta (IL-1beta) were shown previously to upregulate AT(1A) receptor mRNA and protein, thereby increasing the profibrotic response to ANG II in cardiac fibroblasts. The present experiments implicate increased nuclear factor-kappaB (NF-kappaB)-dependent transcription and also, to a lesser extent, altered mRNA splicing in the mechanism of receptor upregulation. Cytokine stimulation was found to increase AT(1A) heterogeneous nuclear RNA levels, which strongly suggests that mRNA upregulation occurs transcriptionally. The transcription factor NF-kappaB was previously deemed necessary for cytokine induced AT(1A) receptor mRNA upregulation. Computer analysis of upstream DNA sequences revealed putative NF-kappaB elements at -365 and -2540 bp. Both isolated elements were shown to bind NF-kappaB (using gel-shift assays) and to transactivate a minimal promoter (using reporter assays), although the element at -365 bp appeared stronger. Three splice variants of AT(1A) receptor mRNA that have different 5' untranslated regions were detected in rat tissues, namely, exons 1-2-3 (predominant), 1-2-3+6, and 1-3. Cytokine treatment of fibroblasts upregulated all splice variants, but exon 1-3 increased more than the others. This differential upregulation, albeit of modest magnitude, was statistically significant with IL-1beta treatment. Exon 2 contains an inhibitory minicistron and a predicted inhibitory hairpin structure. Luciferase reporter assays indicated that each splice variant translates at a different efficiency, with exon 1-2-3+6 (both minicistron and hairpin) < exon 1-2-3 (minicistron only) < exon 1-3 (neither minicistron or hairpin). These results provide evidence that cytokines increase AT(1) protein levels by altering both transcription and splicing. PMID- 15879491 TI - Olmesartan, a novel AT1 antagonist, suppresses cytotoxic myocardial injury in autoimmune heart failure. AB - Some ANG II receptor type 1 (AT(1)) antagonists are reported to inhibit proinflammatory cytokine production in vitro and in vivo. However, the effects of the drugs on autoimmune diseases are unknown. We tested the hypothesis that olmesartan, a novel AT(1) antagonist, ameliorated experimental autoimmune myocarditis (EAM) in rats attributed to the suppression of inflammatory cytokines as well as to the immunomodulatory action of the heart. We administered olmesartan orally at does of 1, 3, and 10 mg.kg(-1).day(-1) to rats with EAM for 3 wk. The results showed that olmesartan decreased blood pressure significantly compared with the untreated group and markedly reduced the severity of myocarditis associated with the decrease of myocardial macrophage, CD4(+), and CD8(+) T-cell expression by comparison of heart wt-to-body wt ratios, pericardial effusion scores, and macroscopic and microscopic scores. Numbers of myocardial interleukin-1beta (IL-1beta)-positive-staining cells (obtained by immunohistochemistry) and quantities of IL-1beta expression (obtained by Western blotting) were significantly lower in rats with EAM given olmesartan treatment compared with rats given vehicle. Cardiac myosin-specific, delayed-type hypersensitivity was significantly lower in olmesartan-treated rats than in control rats. The cytotoxic activities of lymphocytes in rats with EAM treated with olmesartan were reduced compared with untreated control rats. In vitro study showed that both olmesartan and its active metabolite RNH-6270 suppressed IL 1beta production in U-937 cells and cultured myocytes. Olmesartan ameliorates acute EAM in rats. The cardioprotection of olmesartan may be due to suppression of inflammatory cytokines as well as to suppressive effects of cytotoxic myocardial injury in addition to hemodynamic modifications. PMID- 15879492 TI - Trends in antibiotic prescribing for acute respiratory infection in veterans with spinal cord injury and disorder. AB - OBJECTIVES: Most acute respiratory infections (ARIs) are viral and do not warrant antibiotic therapy. Studies to date have not examined trends in antibiotic use in ARIs in populations with disabilities, thus we assessed antibiotic prescribing for veterans with spinal cord injury and disorder (SCI and D) with outpatient ARI visits. PATIENTS AND METHODS: Retrospective study using Department of Veterans Affairs (VA) administrative and pharmacy datasets (1 October 1998-30 September 2001; fiscal years 1999-2001) to assess antibiotic prescribing for upper respiratory infection (URI), lower respiratory infection (LRI), and pneumonia in veterans with SCI and D. RESULTS: There were 5713 ARI visits; 50% received new antibiotic prescriptions. URI and LRI visits were 2.3 times and almost 4 times (P < 0.0001), respectively, more likely to have antibiotics prescribed than pneumonia visits. The majority of URI visits with antibiotic prescriptions had a diagnosis of the common cold or URI not otherwise specified (78%). Acute bronchitis without exacerbation was associated with 95% of LRI visits that received antibiotics. Broad-spectrum antibiotic use increased over time (1999, 46%; 2001, 62%; P < 0.0001). CONCLUSIONS: Although rates of antibiotic prescribing remained stable, prescriptions for broad-spectrum antibiotics increased. Most prescriptions were for indications for which antibiotic use is generally not recommended. Since patients with SCI and D are susceptible to multiple complications, providers may be more concerned with ensuring that any infection is treated, rather than the potential for overuse and resistance. Future efforts should focus on defining benefits of antibiotic use for ARIs in those with disabilities, predictors of prescribing, and interventions to prevent injudicious use of antibiotics. PMID- 15879493 TI - Direct detection in clinical samples of multiple gene mutations causing resistance of Mycobacterium tuberculosis to isoniazid and rifampicin using fluorogenic probes. AB - BACKGROUND: This study evaluates a method based on real-time PCR for direct detection in clinical samples of the common mutations responsible for isoniazid and rifampicin resistance of Mycobacterium tuberculosis. METHODS: Six pairs of fluorogenic 5' exonuclease probes (Taqman), mutated and wild-type, were designed for six targets: codon 315 of katG, substitution C209T in the regulatory region of inhA, and codons 513, 516, 526 and 531 of rpoB. RESULTS: A total of 98 clinical samples harbouring resistant bacilli from 55 patients and 126 samples harbouring susceptible bacilli from 126 patients were processed. The isolates from samples were tested for drug susceptibility with the radiometric method and sequenced for the same genetic targets. Among the samples, 93 harboured isoniazid resistant bacilli. According to the sequencing results, 30 had mutations in katG, 30 in inhA and 33 (35.4%) had no mutations in these targets. All 27 clinical specimens harbouring rifampicin-resistant bacilli showed mutations in rpoB. The detection threshold of this method in detecting target genes in serial dilutions of artificial samples was 1.5 x 10(3) cfu/mL. In clinical samples, the sensitivity ranged from 30.4 to 35.3% for smear-negative samples and from 95.1 to 99.2% for smear-positive samples, with a specificity of 100%. In this study, the overall sensitivity in detecting patients having the target mutations was 74.3%. CONCLUSIONS: The main advantage of the described method is the possibility of detecting rifampicin and isoniazid resistance within 48-72 h after sample collection, with a sensitivity of nearly 100% in smear-positive samples if the chosen target is responsible for the resistance. PMID- 15879494 TI - Characterization of microsomal cytochrome P450-dependent monooxygenases in the rat olfactory mucosa. AB - Nasal administration of a drug ensures therapeutic action by rapid systemic absorption and/or the entry of some molecules into the brain through different routes. Many recent studies have pointed out the presence of xenobiotic metabolizing enzymes in rat olfactory mucosa (OM). Nevertheless, very little is known about the precise identity of isoforms of cytochrome P450 (P450)-dependent monooxygenases (P450) and their metabolic function in this tissue. Therefore, we evaluated mRNA expression of 19 P450 isoforms by semiquantitative reverse transcriptase-polymerase chain reaction and measured their microsomal activity toward six model substrates. For purposes of comparison, studies were conducted on OM and the liver. Specific activities toward phenacetin, chlorzoxazone, and dextromethorphan are higher in OM than in the liver; those toward lauric acid and testosterone are similar in both tissues, and that toward tolbutamide is much lower in OM. There are considerable differences between the two tissues with regard to mRNA expression of P450 isoforms. Some isoforms are expressed in OM but not in the liver (CYP1A1, 2G1, 2B21, and 4B1), whereas mRNA of others (CYP2C6, 2C11, 2D2, 3A1, 3A2, and 4A1) is present only in hepatic tissue. Although expression of CYP1A2, 2A1, 2A3, 2B2, 2D1, 2D4, 2E1, 2J4, and 3A9 is noticed in both tissues, there are a number of quantitative differences. On the whole, our results strongly suggest that CYP1A1, 1A2, 2A3, 2E1, 2G1, and 3A9 are among the main functional isoforms present in OM, at least regarding activities toward the six tested substrates. The implication of olfactory P450-dependent monooxygenases in toxicology, pharmacology, and physiology should be further investigated. PMID- 15879495 TI - Increased bioaccumulation of urethane in CYP2E1-/- versus CYP2E1+/+ mice. AB - Urethane is a fermentation by-product and a potent animal carcinogen. Human exposure to urethane occurs through consumption of alcoholic beverages and fermented foods. Recently, CYP2E1 was identified as the primary enzyme responsible for the metabolism of [(14)C]carbonyl-labeled urethane. Subsequently, attenuation of urethane-induced cell proliferation and genotoxicity in CYP2E1-/- mice was reported. The present work compares the metabolism of single versus multiple exposures of CYP2E1-/- and CYP2E1+/+ mice to (14)C-ethyl-labeled urethane. Urethane was administered as a single 10 or 100 mg/kg gavage dose or at 100 mg/kg/day for 5 consecutive days. CYP2E1+/+ mice administered single or multiple doses exhaled 78 to 88% of dose as (14)CO(2)/day. CYP2E1-/- mice eliminated 30 to 38% of a single dose as (14)CO(2) in 24 h and plateaued after day 3 at approximately 52% of dose/day. The concentrations of urethane-derived radioactivity in plasma and tissues were dose-dependent, increased as a function of the number of doses administered, and were significantly higher in CYP2E1-/- versus CYP2E1+/+ mice. Whereas urethane was the main chemical found in the plasma and tissues of CYP2E1-/- mice, it was not detectable in CYP2E1+/+ mice. In conclusion, multiple dosing led to considerable bioaccumulation of urethane in mice of both genotypes; however, greater retention occurred in CYP2E1-/- versus CYP2E1+/+ mice. Furthermore, greater bioaccumulation of (14)C-ethyl-labeled than [(14)C]carbonyl-labeled urethane was observed in mice. Comparison of the metabolism of ethyl-versus carbonyl-labeled urethane was necessary for tracing the source of CO(2) and led us to propose for the first time that C-hydroxylation is a likely pathway of urethane metabolism. PMID- 15879496 TI - Transendocardial and transepicardial intramyocardial fibroblast growth factor-2 administration: myocardial and tissue distribution. AB - Effective local delivery to the heart remains an obstacle to successful therapeutic application of a number of drugs and biological agents. This study was designed to study and optimize the delivery characteristics of transendocardial intramyocardial (IM) administration, determine myocardial deposition and retention over time, and compare it to transepicardial IM injection. Thirty-nine pigs were used for the study (15 for catheter optimization, 15 for transendocardial IM delivery, and 9 for transepicardial IM delivery). (125)I-Fibroblast growth factor-2 (FGF2) (25 microCi) was used as the prototype molecule. Tissue and myocardial distribution was determined at 1 and 24 h and 7 days. Using 1-h (125)I-FGF2 myocardial deposition as a parameter for delivery efficiency, the optimal needle length and delivery volume for transendocardial based delivery were determined to be 6 mm and 0.1 ml, respectively. Using these parameters for endocardial delivery, (125)I-FGF2 cardiac activity was 18.01 +/- 3.84% of delivered activity at 1 h, 11.65 +/- 5.17% at 24 h, and 2.32 +/- 0.87% at 7 days in ischemic animals. Studies in nonischemic animals produced similar results. For transepicardial delivery, (125)I-FGF2 cardiac-specific activity was 23.14 +/- 12.67% for the 6-mm needle, declining to 12.32 +/- 8.50% at 24 h, and did not significantly differ from values obtained following transendocardial delivery. Thus, optimized transendocardial intramyocardial delivery using Biosense guidance results in efficient delivery of FGF2 to the target myocardium that is comparable with transepicardial delivery, both providing markedly higher myocardial deposition and retention and lower systemic recirculation of FGF2 than intracoronary, intrapericardial, or intravenous delivery. However, myocardial distribution is limited to injection sites. PMID- 15879497 TI - High incidence of skewed X chromosome inactivation in young patients with familial non-BRCA1/BRCA2 breast cancer. AB - BACKGROUND: A higher frequency of skewed X chromosome inactivation has been reported in a consecutive series of young patients with breast cancer compared with controls of a similar age. OBJECTIVE: To investigate the X inactivation pattern in patients with familial non-BRCA1/BRCA2 breast cancer (n = 272), BRCA1/BRCA2 germline mutations (n = 35), and sporadic breast cancer (n = 292). METHODS: X inactivation pattern was determined by polymerase chain reaction analysis of the highly polymorphic CAG repeat in the androgen receptor (AR) gene. The X inactivation pattern was classified as skewed when 90% or more of the cells preferentially expressed one X chromosome. RESULTS: Young patients with familial breast cancer had a significantly higher frequency of skewed X inactivation (11.2%) than young controls (2.7%) (p = 0.001). There was also a strong tendency for middle aged patients with sporadic breast cancer to be more skewed than middle aged controls (13.6% v 4.4%) (p = 0.02). No association between skewed X inactivation and breast cancer was found for the BRCA1/BRCA2 patients . CONCLUSIONS: Skewed X inactivation may be a risk factor for the development of breast cancer in both sporadic and familial breast cancer and may indicate an effect of X linked genes. PMID- 15879498 TI - A common variant of CDKN2A (p16) predisposes to breast cancer. AB - BACKGROUND: A common missense variant of the CDKN2A gene (A148T) predisposes to malignant melanoma in Poland. An association between malignant melanoma and breast cancer has been reported in several families with CDKN2A mutations, OBJECTIVE: To determine whether this variant also predisposes to breast cancer. METHODS: Genotyping was undertaken in 4209 cases of breast cancer, unselected for family history, from 18 hospitals throughout Poland and in 3000 controls. RESULTS: The odds ratio (OR) associated with the CDKN2A allele for women diagnosed with breast cancer before the age of 50 was 1.5 (p = 0.002) and after age 50 it was 1.3 (p = 0.2). The effect was particularly strong for patients diagnosed at or before the age of 30 (OR = 3.8; p = 0.0002). CONCLUSIONS: CDKN2A appears to be a low penetrance breast cancer susceptibility gene in Poland. The association should be confirmed in other populations. PMID- 15879499 TI - Oculocerebrocutaneous syndrome: the brain malformation defines a core phenotype. AB - BACKGROUND: Oculocerebrocutaneous syndrome (OCCS) is characterised by orbital cysts and anophthalmia or microphthalmia, focal aplastic or hypoplastic skin defects, skin appendages, and brain malformations. The eye and skin abnormalities are well described but the neuropathological features less so. To date, 28 patients with an unequivocal diagnosis of OCCS have been reported, with a preponderance of males. OBJECTIVE: To evaluate the brain imaging studies, clinical records, photographs, and pathological material of two new and nine previously reported cases of OCCS. RESULTS: There was a consistent pattern of malformations in eight of the 11 cases, consisting of frontal predominant polymicrogyria and periventricular nodular heterotopia, enlarged lateral ventricles or hydrocephalus, agenesis of the corpus callosum sometimes associated with interhemispheric cysts, and a novel mid-hindbrain malformation. The latter consisted of a giant and dysplastic tectum, absent cerebellar vermis, small cerebellar hemispheres in most cases, and a large posterior fossa fluid collection. CONCLUSIONS: The mid-hindbrain malformation appears pathognomonic for OCCS. The eye and skin features of OCCS show considerable overlap with several other syndromes, such as encephalocraniocutaneous lipomatosis, oculo-auriculo vertebral spectrum, and focal dermal hypoplasia, none of which has a comparable pattern of brain malformations. In particular the unique mid-hindbrain malformation also distinguishes OCCS from related syndromes with comparable forebrain anomalies. The pattern of malformation described thus helps in differentiating OCCS from other entities. The mid-hindbrain malformation points to a defect of the mid-hindbrain organiser as the underlying pathogenic mechanism. PMID- 15879500 TI - Hereditary haemorrhagic telangiectasia: current views on genetics and mechanisms of disease. AB - Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant disorder characterised by epistaxis, telangiectases, and multiorgan vascular dysplasia. The two major types of disease, HHT1 and HHT2, are caused by mutations in the ENG (endoglin) and ACVRL1 genes, respectively. The corresponding endoglin and ALK-1 proteins are specific endothelial receptors of the transforming growth factor beta superfamily essential for maintaining vascular integrity. Many mutations have been identified in ENG and ACVRL1 genes and support the haploinsufficiency model for HHT. Two more genes have recently been implicated in HHT: MADH4 mutated in a combined syndrome of juvenile polyposis and HHT (JPHT), and an unidentified HHT3 gene linked to chromosome 5. Current knowledge on the genetics of HHT is summarised, including the pathways that link the genes responsible for HHT and the potential mechanisms underlying the pathogenesis of the disease. PMID- 15879501 TI - Global analysis of uniparental disomy using high density genotyping arrays. AB - BACKGROUND: Uniparental disomy (UPD), the inheritance of both copies of a chromosome from a single parent, has been identified as the cause for congenital disorders such as Silver-Russell, Prader-Willi, and Angelman syndromes. Detection of UPD has largely been performed through labour intensive screening of DNA from patients and their parents, using microsatellite markers. METHODS: We applied high density single nucleotide polymorphism (SNP) microarrays to diagnose whole chromosome and segmental UPD and to study the occurrence of continuous or interspersed heterodisomic and isodisomic regions in six patients with Silver Russell syndrome patients who had maternal UPD for chromosome 7 (matUPD7). RESULTS: We have devised a new high precision and high-throughput computational method to confirm UPD and to localise segments where transitions of UPD status occur. Our method reliably confirmed and mapped the matUPD7 regions in all patients in our study. CONCLUSION: Our results suggest that high density SNP arrays can be reliably used for rapid and efficient diagnosis of both segmental and whole chromosome UPD across the entire genome. PMID- 15879502 TI - Primary pulmonary synovial sarcoma confirmed by molecular detection of SYT-SSX1 fusion gene transcripts: a case report and review of the literature. AB - This is a case report of a rare patient with primary pulmonary synovial sarcoma. The patient was a 58-year-old woman who presented with a well-defined giant mass in the right lower field on a chest radiograph. A malignant pulmonary tumor was suspected and consequently a right middle and lower lobectomy was performed. Grossly, the tumor measured 10 x 8 x 7 cm, was whitish-yellow in color and friable with hemorrhage. Histologically, the tumor showed a dense proliferation of spindle cells. In some areas, a herringbone-like pattern with coagulation necrosis of large size was noted. Immunohistochemically, the tumor cells were focally positive for cytokeratin and epithelial membrane antigen (EMA). As these features suggested a monophasic synovial sarcoma, we looked for the presence of SYT-SSX fusion gene transcripts using RNA samples from the paraffin-embedded tissue. A reverse transcription-polymerase chain reaction (RT-PCR) amplified a single 118 bp fragment characteristic of the SYT-SSX1 fusion gene transcripts. As no tumor was found at other sites, it was diagnosed as primary pulmonary synovial sarcoma. Molecular testing proved to be very helpful or necessary when monophasic spindle cell synovial sarcoma was recognized in uncommon/unexpected sites. In our review of primary pulmonary synovial sarcomas confirmed by molecular detection of SYT-SSX fusion gene transcripts, the SYT-SSX2 fusion protein expression correlates with poorer prognosis. This is in contrast to the association between the SYT-SSX1 fusion protein expression and poorer prognosis in soft tissue synovial sarcomas. PMID- 15879503 TI - The molecular chaperone Sse1 and the growth control protein kinase Sch9 collaborate to regulate protein kinase A activity in Saccharomyces cerevisiae. AB - The Sch9 protein kinase regulates Hsp90-dependent signal transduction activity in the budding yeast Saccharomyces cerevisiae. Hsp90 functions in concert with a number of cochaperones, including the Hsp110 homolog Sse1. In this report, we demonstrate a novel synthetic genetic interaction between SSE1 and SCH9. This interaction was observed specifically during growth at elevated temperature and was suppressed by decreased signaling through the protein kinase A (PKA) signal transduction pathway. Correspondingly, sse1Delta sch9Delta cells were shown by both genetic and biochemical approaches to have abnormally high levels of PKA activity and were less sensitive to modulation of PKA by glucose availability. Growth defects of an sse1Delta mutant were corrected by reducing PKA signaling through overexpression of negative regulators or growth on nonoptimal carbon sources. Hyperactivation of the PKA pathway through expression of a constitutive RAS2 allele likewise resulted in temperature-sensitive growth, suggesting that modulation of PKA activity during thermal stress is required for adaptation and viability. Together these results demonstrate that the Sse1 chaperone and the growth control kinase Sch9 independently contribute to regulation of PKA signaling. PMID- 15879504 TI - The inactivation of KlNOT4, a Kluyveromyces lactis gene encoding a component of the CCR4-NOT complex, reveals new regulatory functions. AB - We have isolated the KlNOT4 gene of the yeast Kluyveromyces lactis, which encodes a component of the evolutionarily conserved CCR4-NOT complex. We show that inactivation of the gene leads to pleiotropic defects that were differentially suppressed by the NOT4 gene of S. cerevisiae, indicating that these genes have overlapping, but not identical, functions. K. lactis strains lacking Not4p are defective in fermentation and show reduced transcription of glucose transporter and glycolytic genes, which are phenotypes that are not found in the corresponding mutant of S. cerevisiae. We also show that Not4 proteins control the respiratory pathway in both yeasts, although with some differences. They activate transcription of KlACS2 and KlCYC1, but repress KlICL1, ScICL1, ScACS1, and ScCYC1. Altogether, our results indicate that Not4p is a pivotal factor involved in the regulation of carbon metabolism in yeast. PMID- 15879506 TI - The origin of multiple B mating specificities in Coprinus cinereus. AB - Mushrooms, such as Coprinus cinereus, possess large families of pheromones and G protein-coupled receptors that are sequestered at the B mating-type locus and whose function is to confer vast numbers of different mating types. This ability results from complex patterns of cognate and noncognate pheromone/receptor pairings, which potentially offer a unique insight into the molecular interaction between receptor and ligand. In this study we have identified many more members of these families by molecular analysis of strains collected worldwide. There are three groups of genes at each B locus. We have identified two alleles of group 1, five alleles of group 2, and seven alleles of group 3, encoding in total 14 different receptors and 29 different pheromones. The specificity of many newly identified alleles was determined by transformation analysis. One striking finding was that receptors fall into groups based on sequence homology but these do not correspond to the groups defined by position, indicating that complex evolutionary processes gave rise to the B loci. While additional allelic versions may occur in nature, the number of B specificities possible by combination of the alleles that we describe is 70, close to previous estimates based on population analysis. PMID- 15879507 TI - Epistasis and the adaptability of an RNA virus. AB - We have explored the patterns of fitness recovery in the vesicular stomatitis RNA virus. We show that, in our experimental setting, reversions to the wild-type genotype were rare and fitness recovery was at least partially driven by compensatory mutations. We compared compensatory adaptation for genotypes carrying (1) mutations with varying deleterious fitness effects, (2) one or two deleterious mutations, and (3) pairs of mutations showing differences in the strength and sign of epistasis. In all cases, we found that the rate of fitness recovery and the proportion of reversions were positively affected by population size. Additionally, we observed that mutations with large fitness effect were always compensated faster than mutations with small fitness effect. Similarly, compensatory evolution was faster for genotypes carrying a single deleterious mutation than for those carrying pairs of mutations. Finally, for genotypes carrying two deleterious mutations, we found evidence of a negative correlation between the epistastic effect and the rate of compensatory evolution. PMID- 15879508 TI - Comparison of the X and Y chromosome organization in Silene latifolia. AB - Here we compare gene orders on the Silene latifolia sex chromosomes. On the basis of the deletion mapping results (11 markers and 23 independent Y chromosome deletion lines used), we conclude that a part of the Y chromosome (covering a region corresponding to at least 23.9 cM on the X chromosome) has been inverted. The gradient in silent-site divergence suggests that this inversion took place after the recombination arrest in this region. Because recombination arrest events followed by Y chromosome rearrangements also have been found in the human Y chromosome, this process seems to be a general evolutionary pathway. PMID- 15879505 TI - Pericentromeric regions of soybean (Glycine max L. Merr.) chromosomes consist of retroelements and tandemly repeated DNA and are structurally and evolutionarily labile. AB - Little is known about the physical makeup of heterochromatin in the soybean (Glycine max L. Merr.) genome. Using DNA sequencing and molecular cytogenetics, an initial analysis of the repetitive fraction of the soybean genome is presented. BAC 076J21, derived from linkage group L, has sequences conserved in the pericentromeric heterochromatin of all 20 chromosomes. FISH analysis of this BAC and three subclones on pachytene chromosomes revealed relatively strict partitioning of the heterochromatic and euchromatic regions. Sequence analysis showed that this BAC consists primarily of repetitive sequences such as a 102-bp tandem repeat with sequence identity to a previously characterized approximately 120-bp repeat (STR120). Fragments of Calypso-like retroelements, a recently inserted SIRE1 element, and a SIRE1 solo LTR were present within this BAC. Some of these sequences are methylated and are not conserved outside of G. max and G. soja, a close relative of soybean, except for STR102, which hybridized to a restriction fragment from G. latifolia. These data present a picture of the repetitive fraction of the soybean genome that is highly concentrated in the pericentromeric regions, consisting of rapidly evolving tandem repeats with interspersed retroelements. PMID- 15879510 TI - Efficiency of the use of pedigree and molecular marker information in conservation programs. AB - The value of molecular markers and pedigree records, separately or in combination, to assist in the management of conserved populations has been tested. The general strategy for managing the population was to optimize contributions of parents to the next generation for minimizing the global weighted coancestry. Strategies differed in the type of information used to compute global coancestries, the number and type of evaluated individuals, and the system of mating. Genealogical information proved to be very useful (at least for 10 generations of management) to arrange individuals' contributions via the minimization of global coancestry. In fact, the level of expected heterozygosity after 10 generations yielded by this strategy was 88-100% of the maximum possible improvement obtained if the genotype for all loci was known. Marker information was of very limited value if used alone. The amount and degree of polymorphism of markers to be used to compute molecular coancestry had to be high to mimic the performance of the strategy relying on pedigree, especially in the short term (for example, >10 markers per chromosome with 10 alleles each were needed if only the parents' genotype was available). When both sources of information are combined to calculate the coancestry conditional on markers, clear increases in effective population size (Ne) were found, but observed diversity levels (either gene or allelic diversity) in the early generations were quite similar to the ones obtained with pedigree alone. The advantage of including molecular information is greater when information is available on a greater number of individuals (offspring and parents vs. parents only). However, for realistic situations (i.e., large genomes) the benefits of using information on offspring are small. The same conclusions were reached when comparing the use of the different types of information (genealogical or/and molecular) to perform minimum coancestry matings. PMID- 15879509 TI - The genetic basis of resistance to anticoagulants in rodents. AB - Anticoagulant compounds, i.e., derivatives of either 4-hydroxycoumarin (e.g., warfarin, bromadiolone) or indane-1,3-dione (e.g., diphacinone, chlorophacinone), have been in worldwide use as rodenticides for >50 years. These compounds inhibit blood coagulation by repression of the vitamin K reductase reaction (VKOR). Anticoagulant-resistant rodent populations have been reported from many countries and pose a considerable problem for pest control. Resistance is transmitted as an autosomal dominant trait although, until recently, the basic genetic mutation was unknown. Here, we report on the identification of eight different mutations in the VKORC1 gene in resistant laboratory strains of brown rats and house mice and in wild-caught brown rats from various locations in Europe with five of these mutations affecting only two amino acids (Tyr139Cys, Tyr139Ser, Tyr139Phe and Leu128Gln, Leu128Ser). By recombinant expression of VKORC1 constructs in HEK293 cells we demonstrate that mutations at Tyr139 confer resistance to warfarin at variable degrees while the other mutations, in addition, dramatically reduce VKOR activity. Our data strongly argue for at least seven independent mutation events in brown rats and two in mice. They suggest that mutations in VKORC1 are the genetic basis of anticoagulant resistance in wild populations of rodents, although the mutations alone do not explain all aspects of resistance that have been reported. We hypothesize that these mutations, apart from generating structural changes in the VKORC1 protein, may induce compensatory mechanisms to maintain blood clotting. Our findings provide the basis for a DNA-based field monitoring of anticoagulant resistance in rodents. PMID- 15879511 TI - The coupon collector and the suppressor mutation: estimating the number of compensatory mutations by maximum likelihood. AB - Compensatory mutation occurs when a loss of fitness caused by a deleterious mutation is restored by its epistatic interaction with a second mutation at a different site in the genome. How many different compensatory mutations can act on a given deleterious mutation? Although this quantity is fundamentally important to understanding the evolutionary consequence of mutation and the genetic complexity of adaptation, it remains poorly understood. To determine the shape of the statistical distribution for the number of compensatory mutations per deleterious mutation, we have performed a maximum-likelihood analysis of experimental data collected from the suppressor mutation literature. Suppressor mutations are used widely to assess protein interactions and are under certain conditions equivalent to compensatory mutations. By comparing the maximum likelihood of a variety of candidate distribution functions, we established that an L-shaped gamma distribution (alpha=0.564, theta=21.01) is the most successful at explaining the collected data. This distribution predicts an average of 11.8 compensatory mutations per deleterious mutation. Furthermore, the success of the L-shaped gamma distribution is robust to variation in mutation rates among sites. We have detected significant differences among viral, prokaryotic, and eukaryotic data subsets in the number of compensatory mutations and also in the proportion of compensatory mutations that are intragenic. This is the first attempt to characterize the overall diversity of compensatory mutations, identifying a consistent and accurate prior distribution of compensatory mutation diversity for theoretical evolutionary models. PMID- 15879512 TI - Quantitative trait locus analysis using recombinant inbred intercrosses: theoretical and empirical considerations. AB - We describe a new approach, called recombinant inbred intercross (RIX) mapping, that extends the power of recombinant inbred (RI) lines to provide sensitive detection of quantitative trait loci (QTL) responsible for complex genetic and nongenetic interactions. RIXs are generated by producing F1 hybrids between all or a subset of parental RI lines. By dramatically extending the number of unique, reproducible genomes, RIXs share some of the best properties of both the parental RI and F2 mapping panels. These attributes make the RIX method ideally suited for experiments requiring analysis of multiple parameters, under different environmental conditions and/or temporal sampling. However, since any pair of RIX genomes shares either one or no parental RIs, this cross introduces an unusual population structure requiring special computational approaches for analysis. Herein, we propose an efficient statistical procedure for QTL mapping with RIXs and describe a novel empirical permutation procedure to assess genome-wide significance. This procedure will also be applicable to diallel crosses. Extensive simulations using strain distribution patterns from CXB, AXB/BXA, and BXD mouse RI lines show the theoretical power of the RIX approach and the analysis of CXB RIXs demonstrates the limitations of this procedure when using small RI panels. PMID- 15879513 TI - A composite-likelihood approach for detecting directional selection from DNA sequence data. AB - We present a novel composite-likelihood-ratio test (CLRT) for detecting genes and genomic regions that are subject to recurrent natural selection (either positive or negative). The method uses the likelihood functions of Hartl et al. (1994) for inference in a Wright-Fisher genic selection model and corrects for nonindependence among sites by application of coalescent simulations with recombination. Here, we (1) characterize the distribution of the CLRT statistic (Lambda) as a function of the population recombination rate (R=4Ner); (2) explore the effects of bias in estimation of R on the size (type I error) of the CLRT; (3) explore the robustness of the model to population growth, bottlenecks, and migration; (4) explore the power of the CLRT under varying levels of mutation, selection, and recombination; (5) explore the discriminatory power of the test in distinguishing negative selection from population growth; and (6) evaluate the performance of maximum composite-likelihood estimation (MCLE) of the selection coefficient. We find that the test has excellent power to detect weak negative selection and moderate power to detect positive selection. Moreover, the test is quite robust to bias in the estimate of local recombination rate, but not to certain demographic scenarios such as population growth or a recent bottleneck. Last, we demonstrate that the MCLE of the selection parameter has little bias for weak negative selection and has downward bias for positively selected mutations. PMID- 15879515 TI - Cornfests, cornfabs and cooperation: the origins and beginnings of the maize genetics cooperation news letter. PMID- 15879514 TI - The large loop repair and mismatch repair pathways of Saccharomyces cerevisiae act on distinct substrates during meiosis. AB - During meiotic recombination in the yeast Saccharomyces cerevisiae, heteroduplex DNA is formed when single-stranded DNAs from two homologs anneal as a consequence of strand invasion. If the two DNA strands differ in sequence, a mismatch will be generated. Mismatches in heteroduplex DNA are recognized and repaired efficiently by meiotic DNA mismatch repair systems. Components of two meiotic systems, mismatch repair (MMR) and large loop repair (LLR), have been identified previously, but the substrate range of these repair systems has never been defined. To determine the substrates for the MMR and LLR repair pathways, we constructed insertion mutations at HIS4 that form loops of varying sizes when complexed with wild-type HIS4 sequence during meiotic heteroduplex DNA formation. We compared the frequency of repair during meiosis in wild-type diploids and in diploids lacking components of either MMR or LLR. We find that the LLR pathway does not act on single-stranded DNA loops of <16 nucleotides in length. We also find that the MMR pathway can act on loops up to 17, but not >19, nucleotides in length, indicating that the two pathways overlap slightly in their substrate range during meiosis. Our data reveal differences in mitotic and meiotic MMR and LLR; these may be due to alterations in the functioning of each complex or result from subtle sequence context influences on repair of the various mismatches examined. PMID- 15879516 TI - A major species of mouse mu-opioid receptor mRNA and its promoter-dependent functional polyadenylation signal. AB - The pharmacological effects of opioid drugs are mediated mainly by the mu-opioid receptor (MOR), which is encoded by an mRNA transcript named MOR1. Although several MOR mRNA splice variants have been reported, their biological relevance has been debated. In this study, we found that probes of regions essential for the production of functional MOR, as well as that of the 3'-downstream region of the MOR gene coding region, detected by Northern blot analyses, a major species of mature transcript MOR1 from mouse brain of approximately 11.5 kilobases (kb). Although exon 3 probe detected an additional 3.7-kb transcript, this transcript was not detected by other probes, ruling out its ability to produce functional MOR. The 3'-untranslated region (UTR) of MOR1 is contiguously extended from the end of the coding region, and uses a single polyadenylation [poly (A)] signal (located 10,179 bp downstream of the MOR1 stop codon). The poly (A) signal (AAUAAA) is located 26 bp upstream of the poly (A) site. Transient transfection using luciferase reporters verified the functionality of this poly (A) signal, in particular on a reporter driven by the MOR promoter. This poly (A) is much less effective for a heterologous promoter, such as simian virus 40, indicating a functional coupling of MOR promoter and its own poly (A). This report verifies MOR1 as the major mature MOR gene transcript that has the full capacity to produce functional MOR protein, identifies the 3'-UTR of MOR1 transcript, and uncovers functional coupling of the MOR gene promoter and its polyadenylation signal. PMID- 15879517 TI - Down-regulation of Na+/Ca2+ exchanger by fluvastatin in rat cardiomyoblast H9c2 cells: involvement of RhoB in Na+/Ca2+ exchanger mRNA stability. AB - We investigated the effect of fluvastatin (Flv), an HMG-CoA reductase inhibitor, on Na(+)/Ca(2+) exchanger 1 (NCX1) expression in H9c2 cardiomyoblasts. Reverse transcriptase-polymerase chain reaction analyses revealed that Flv decreased NCX1 mRNA in a concentration- and time-dependent manner and NCX1 protein. This effect of Flv was caused by the inhibition of HMG-CoA reductase, because Flv did not affect the NCX1 mRNA in the presence of mevalonate. Flv-induced down-regulation of NCX1 mRNA was also cancelled by farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), suggesting an involvement of small G proteins. However, overexpression of neither constitutive active RhoA nor Ras affected NCX1 mRNA. In contrast, intracellular expression of C3 toxin, a specific inhibitor of Rho family proteins, decreased NCX1 mRNA, suggesting that Flv decreases NCX1 mRNA by inhibiting a signaling pathway of Rho family proteins other than RhoA. On the other hand, lysophosphatidylcholine (LPC), an activator of Rho signaling, increased both NCX1 mRNA and protein in a C3 toxin-sensitive manner. Western blot analyses revealed that membrane-associated RhoB, which is isoprenylated by either FPP or GGPP, was decreased by Flv but was increased by LPC. Selective inhibition of gene expression by short interfering RNA duplex showed that RhoB but not RhoA is involved in the regulation of NCX1 mRNA and protein. When transcription was blocked by 5,6-dichlorobenzimidazole riboside, the NCX1 mRNA stability was decreased by Flv. Long-term treatment of rat with Flv in vivo also down-regulated the cardiac NCX1 mRNA. These results suggest that a RhoB-mediated signaling pathway regulates cardiac NCX1 levels by controlling the NCX1 mRNA stability. PMID- 15879518 TI - The zrfA and zrfB genes of Aspergillus fumigatus encode the zinc transporter proteins of a zinc uptake system induced in an acid, zinc-depleted environment. AB - Zinc is an essential micronutrient that cells must obtain from the environment in order to develop their normal growth. Previous work performed at our laboratory showed that the synthesis of immunodominant antigens from Aspergillus spp., including A. fumigatus, was up-regulated by a low environmental concentration of zinc. These results suggested that a tightly regulated system for the fungus to grow under zinc-limiting conditions must underlie the ability of A. fumigatus to acquire zinc in such environments. In this work, we show that zrfA and zrfB are two of the genes that encode membrane zinc transporters from A. fumigatus in this system. Expression of these genes is differentially down-regulated by increasing concentrations of zinc in the medium. Thus, the transcription of zrfB is turned off at a concentration 50-fold higher than that for zrfA transcription. In addition, phenotypic analyses of single zrfADelta and zrfBDelta mutants and a double zrfAzrfBDelta mutant revealed that the deletion of zrfB causes a greater defect in growth than the single deletion of zrfA. Deletion of both genes has a dramatic effect on growth under acid, zinc-limiting conditions. Interestingly, in neutral or slightly alkaline zinc-depleted medium, the transcriptional expression of both genes is down-regulated to such an extent that even in the absence of a supplement of zinc, the expression of zrfA and zrfB is strongly reduced. This fact correlates with the growth observed in alkaline medium, in which even a zrfAzrfBDelta double mutant was able to grow in a similar way to the wild-type under extremely zinc-limiting conditions. In sum, the zinc transport proteins encoded by zrfA and zrfB are members of a zinc uptake system of A. fumigatus that operates mainly under acid, zinc-limiting conditions. PMID- 15879519 TI - A two-hybrid screen of the yeast proteome for Hsp90 interactors uncovers a novel Hsp90 chaperone requirement in the activity of a stress-activated mitogen activated protein kinase, Slt2p (Mpk1p). AB - The Hsp90 chaperone cycle catalyzes the final activation step of several important eukaryotic proteins (Hsp90 "clients"). Although largely a functional form of Hsp90, an Hsp90-Gal4p DNA binding domain fusion (Hsp90-BD) displays no strong interactions in the yeast two-hybrid system, consistent with a general transience of most Hsp90-client associations. Strong in vivo interactions are though detected when the E33A mutation is introduced into this bait, a mutation that should arrest Hsp90-client complexes at a stage where the client is stabilized, yet prevented from attaining its active form. This E33A mutation stabilized the two-hybrid interactions of the Hsp90-BD fusion with approximately 3% of the Saccharomyces cerevisiae proteome in a screen of the 6,000 yeast proteins expressed as fusions to the Gal4p activation domain (AD). Among the detected interactors were the two stress-activated mitogen-activated protein (MAP) kinases of yeast, Hog1p and Slt2p (Mpk1p). Column retention experiments using wild-type and mutant forms of Hsp90 and Slt2p MAP kinase, as well as quantitative measurements of the effects of stress on the two-hybrid interaction of mutant Hsp90-BD and AD-Slt2p fusions, revealed that Hsp90 binds exclusively to the dually Thr/Tyr-phosphorylated, stress-activated form of Slt2p [(Y-P,T P)Slt2p] and also to the MAP kinase domain within this (Y-P,T-P)Slt2p. Phenotypic analysis of a yeast mutant that expresses a mutant Hsp90 (T22Ihsp82) revealed that Hsp90 function is essential for this (Y-P,T-P)Slt2p to activate one of its downstream targets, the Rlm1p transcription factor. The interaction between Hsp90 and (Y-P,T-P)Slt2p, characterized in this study, is probably essential in this Hsp90 facilitation of the Rlm1p activation by Slt2p. PMID- 15879520 TI - Role of Tos3, a Snf1 protein kinase kinase, during growth of Saccharomyces cerevisiae on nonfermentable carbon sources. AB - In Saccharomyces cerevisiae, Snf1 protein kinase of the Snf1/AMP-activated protein kinase family is required for growth on nonfermentable carbon sources and nonpreferred sugars. Three kinases, Pak1, Elm1, and Tos3, activate Snf1 by phosphorylation of its activation-loop threonine, and the absence of all three causes the Snf(-) phenotype. No phenotype has previously been reported for the tos3Delta single mutation. We show here that, when cells are grown on glycerol ethanol, tos3Delta reduces growth rate, Snf1 catalytic activity, and activation of the Snf1-dependent carbon source-responsive element (CSRE) in the promoters of gluconeogenic genes. In contrast, tos3Delta did not significantly affect Snf1 catalytic activity or CSRE function during abrupt glucose depletion, indicating that Tos3 has a more substantial role in activating Snf1 protein kinase during growth on a nonfermentable carbon source than during acute carbon stress. We also report that Tos3 is localized in the cytosol during growth in either glucose or glycerol-ethanol. These findings lend support to the idea that the Snf1 protein kinase kinases make different contributions to cellular regulation under different growth conditions. PMID- 15879521 TI - Two complexes of spindle checkpoint proteins containing Cdc20 and Mad2 assemble during mitosis independently of the kinetochore in Saccharomyces cerevisiae. AB - Favored models of spindle checkpoint signaling propose that two inhibitory complexes (Mad2-Cdc20 and Mad2-Mad3-Bub3-Cdc20) must be assembled at kinetochores in order to inhibit mitosis. We have directly tested this model in the budding yeast Saccharomyces cerevisiae. The proteins Mad2, Mad3, Bub3, Cdc20, and Cdc27 in yeast were quantified, and there are sufficient amounts to form stoichiometric inhibitors of Cdc20 and the anaphase-promoting complex. Mad2 is present in two separate complexes in cells arrested in mitosis with nocodazole. There is a small amount of Mad2-Mad3-Bub3-Cdc20 and a much larger amount of a complex that contains Mad2-Cdc20. We use conditional mutants to show that both Mad2 and Mad3 are essential for establishment and maintenance of the spindle checkpoint. Both spindle checkpoint complexes containing Mad2 form in mitosis, not in response to checkpoint activation. The kinetochore is not required to form either complex. We propose that the conversion of Mad1-Mad2 to Cdc20-Mad2, a key step in generating inhibitory checkpoint complexes, is limited to mitosis by the availability of Cdc20 and is kinetochore independent. PMID- 15879522 TI - Mannosyl-diinositolphospho-ceramide, the major yeast plasma membrane sphingolipid, governs toxicity of Kluyveromyces lactis zymocin. AB - Kluyveromyces lactis zymocin, a trimeric (alphabetagamma) protein toxin complex, inhibits proliferation of Saccharomyces cerevisiae cells. Here we present an analysis of kti6 mutants, which resist exogenous zymocin but are sensitive to intracellular expression of its inhibitory gamma-toxin subunit, suggesting that KTI6 encodes a factor needed for toxin entry into the cell. Consistent with altered cell surface properties, kti6 cells resist hygromycin B, syringomycin E, and nystatin, antibiotics that require intact membrane potentials or provoke membrane disruption. KTI6 is allelic to IPT1, coding for mannosyl diinositolphospho-ceramide [M(IP)(2)C] synthase, which produces M(IP)(2)C, the major plasma membrane sphingolipid. kti6 membranes lack M(IP)(2)C and sphingolipid mutants that have reduced levels of M(IP)(2)C precursors, including the sphingolipid building block ceramide survive zymocin. In addition, kti6/ipt1 cells allow zymocin docking but prevent import of its toxic gamma-subunit. Genetic analysis indicates that Kti6 is likely to act upstream of lipid raft proton pump Kti10/Pma1, a previously identified zymocin sensitivity factor. In sum, M(IP)(2)C operates in a plasma membrane step that follows recognition of cell wall chitin by zymocin but precedes the involvement of elongator, the potential toxin target. PMID- 15879523 TI - Relationship of DFG16 to the Rim101p pH response pathway in Saccharomyces cerevisiae and Candida albicans. AB - Many fungal pH responses depend upon conserved Rim101p/PacC transcription factors, which are activated by C-terminal proteolytic processing. The means by which environmental pH is sensed by this pathway are not known. Here, we report a screen of the Saccharomyces cerevisiae viable deletion mutant library that has yielded a new gene required for processed Rim101p accumulation, DFG16. An S. cerevisiae dfg16Delta mutant expresses Rim101p-repressed genes at elevated levels. In addition, Candida albicans dfg16Delta/dfg16Delta mutants are defective in alkaline pH-induced filamentation, and their defect is suppressed by expression of truncated Rim101-405p. Thus, Dfg16p is a functionally conserved Rim101p pathway member. Many proteins required for processed Rim101p accumulation are members of the ESCRT complex, which functions in the formation of multivesicular bodies (MVBs). Staining with the dye FM4-64 indicates that the S. cerevisiae dfg16Delta mutant does not have an MVB defect. We find that two transcripts, PRY1 and ASN1, respond to mutations that affect both the Rim101p and MVB pathways but not to mutations that affect only one pathway. The S. cerevisiae dfg16Delta mutation does not affect PRY1 and ASN1 expression, thus confirming that Dfg16p function is restricted to the Rim101p pathway. Dfg16p is homologous to Aspergillus nidulans PalH, a component of the well-characterized PacC processing pathway. We verify that the previously recognized PalH homolog, Rim21p, also functions in the S. cerevisiae Rim101p pathway. Dfg16p is predicted to have seven membrane-spanning segments and a long hydrophilic C-terminal region, as expected if Dfg16p were a G-protein-coupled receptor. PMID- 15879524 TI - Molecular and functional analyses of poi-2, a novel gene highly expressed in sexual and perithecial tissues of Neurospora crassa. AB - The poi-2 gene is highly and specifically expressed in starved and sexual tissues of the filamentous fungus Neurospora crassa. It encodes a 27-kDa protein, as shown by in vitro transcription and translation. The POI2 protein contains a hydrophobic signal sequence at the amino terminus followed by novel 16 tandem repeats of 13 to 14 amino acid residues; all repeats are separated by Kex2 processing sites. Repeat-induced point mutation (RIP)-mediated gene disruption was used to generate poi-2 mutants, and the mutated sequences showed either one of two distinct patterns: typical RIPs (GC-to-AT transitions) or insertion deletion (indel) mutations. Although the poi-2 strains contained numerous mutations, all retained intact open reading frames (ORFs) of various lengths. They showed greatly reduced vegetative growth and protoperithecial formation and low viability of their sexual progeny. All poi-2 mutants had similar defects in male fertility and the mating response, but the nature of female fertility defects varied and corresponded to the length of the residual poi-2 ORF. Mutants with ORFs of approximately normal length occasionally completed sexual development and produced viable ascospores, while a mutant with a severely truncated ORF was female sterile due to its inability to form protoperithecia. Thus, poi-2 is essential for differentiation of female reproductive structures and perithecial development as well as for normal vegetative growth. The POI2 protein is involved in the mating response, probably as a component in the pathway rather than as a pheromone. PMID- 15879525 TI - Cell biology of conidial anastomosis tubes in Neurospora crassa. AB - Although hyphal fusion has been well documented in mature colonies of filamentous fungi, it has been little studied during colony establishment. Here we show that specialized hyphae, called conidial anastomosis tubes (CATs), are produced by all types of conidia and by conidial germ tubes of Neurospora crassa. The CAT is shown to be a cellular element that is morphologically and physiologically distinct from a germ tube and under separate genetic control. In contrast to germ tubes, CATs are thinner, shorter, lack branches, exhibit determinate growth, and home toward each other. Evidence for an extracellular CAT inducer derived from conidia was obtained because CAT formation was reduced at low conidial concentrations. A cr-1 mutant lacking cyclic AMP (cAMP) produced CATs, indicating that the inducer is not cAMP. Evidence that the transduction of the CAT inducer signal involves a putative transmembrane protein (HAM-2) and the MAK-2 and NRC-1 proteins of a mitogen-activated protein kinase signaling pathway was obtained because ham-2, mak-2, and nrc-1 mutants lacked CATs. Optical tweezers were used in a novel experimental assay to micromanipulate whole conidia and germlings to analyze chemoattraction between CATs during homing. Strains of the same and opposite mating type were shown to home toward each other. The cr-1 mutant also underwent normal homing, indicating that cAMP is not the chemoattractant. ham-2, mak-2, and nrc-1 macroconidia did not attract CATs of the wild type. Fusion between CATs of opposite mating types was partially inhibited, providing evidence of non-self-recognition prior to fusion. Microtubules and nuclei passed through fused CATs. PMID- 15879526 TI - The so locus is required for vegetative cell fusion and postfertilization events in Neurospora crassa. AB - The process of cell fusion is a basic developmental feature found in most eukaryotic organisms. In filamentous fungi, cell fusion events play an important role during both vegetative growth and sexual reproduction. We employ the model organism Neurospora crassa to dissect the mechanisms of cell fusion and cell-cell communication involved in fusion processes. In this study, we characterized a mutant with a mutation in the gene so, which exhibits defects in cell fusion. The so mutant has a pleiotropic phenotype, including shortened aerial hyphae, an altered conidiation pattern, and female sterility. Using light microscopy and heterokaryon tests, the so mutant was shown to possess defects in germling and hyphal fusion. Although so produces conidial anastomosis tubes, so germlings did not home toward wild-type germlings nor were wild-type germlings attracted to so germlings. We employed a trichogyne attraction and fusion assay to determine whether the female sterility of the so mutant is caused by impaired communication or fusion failure between mating partners. so showed no defects in attraction or fusion between mating partners, indicating that so is specific for vegetative hyphal fusion and/or associated communication events. The so gene encodes a protein of unknown function, but which contains a WW domain; WW domains are predicted to be involved in protein-protein interactions. Database searches showed that so was conserved in the genomes of filamentous ascomycete fungi but was absent in ascomycete yeast and basidiomycete species. PMID- 15879527 TI - A Hydrophobin of the chestnut blight fungus, Cryphonectria parasitica, is required for stromal pustule eruption. AB - Hydrophobins are abundant small hydrophobic proteins that are present on the surfaces of many filamentous fungi. The chestnut blight pathogen Cryphonectria parasitica was shown to produce a class II hydrophobin, cryparin. Cryparin is the most abundant protein produced by this fungus when grown in liquid culture. When the fungus is growing on chestnut trees, cryparin is found only in the fungal fruiting body walls. Deletion of the gene encoding cryparin resulted in a culture phenotype typical of hydrophobin deletion mutants of other fungi, i.e., easily wettable (nonhydrophobic) hyphae. When grown on the natural substrate of the fungus, however, cryparin-null mutation strains were unable to normally produce its fungal fruiting bodies. Although the stromal pustules showed normal development initially, they were unable to erupt through the bark of the tree. The hydrophobin cryparin thus plays an essential role in the fitness of this important plant pathogen by facilitating the eruption of the fungal fruiting bodies through the bark of its host tree. PMID- 15879528 TI - Developmental and transcriptional responses to host and nonhost cuticles by the specific locust pathogen Metarhizium anisopliae var. acridum. AB - Transcript patterns elicited in response to hosts can reveal how fungi recognize suitable hosts and the mechanisms involved in pathogenicity. These patterns could be fashioned by recognition of host-specific topographical features or by chemical components displayed or released by the host. We investigated this in the specific locust pathogen Metarhizium anisopliae var. acridum. Only host (Schistocerca gregaria) cuticle stimulated the full developmental program of germination and differentiation of infection structures (appressoria). Cuticle from beetles (Leptinotarsa decimlineata) repressed germination while cuticle from hemipteran bugs (Magicicada septendecim) allowed germination but only very low levels of differentiation, indicating that the ability to cause disease can be blocked at different stages. Using organic solvents to extract insects we identified a polar fraction from locusts that allowed appressorial formation against a flat plastic (hydrophobic) surface. Microarrays comprising 1,730 expressed sequence tags were used to determine if this extract elicits different transcriptional programs than whole locust cuticle or nonhost extracts. Of 483 differentially regulated genes, 97% were upregulated. These included genes involved in metabolism, utilization of host cuticle components, cell survival and detoxification, and signal transduction. Surprisingly, given the complex nature of insect epicuticle components and the specific response of M. anisopliae var. acridum to locusts, very similar transcript profiles were observed on locust and beetle extracts. However, the beetle extract cluster was enriched in genes for detoxification and redox processes, while the locust extract upregulated more genes for cell division and accumulation of cell mass. In addition, several signal transduction genes previously implicated in pathogenicity in plant pathogens were only upregulated in response to locust extract, implying similarities in the regulatory circuitry of these pathogens with very different hosts. PMID- 15879529 TI - Identification in the ancient protist Giardia lamblia of two eukaryotic translation initiation factor 4E homologues with distinctive functions. AB - Eukaryotic translation initiation factor 4E (eIF4E) binds to the m(7)GTP of capped mRNAs and is an essential component of the translational machinery that recruits the 40S small ribosomal subunit. We describe here the identification and characterization of two eIF4E homologues in an ancient protist, Giardia lamblia. Using m(7)GTP-Sepharose affinity column chromatography, a specific binding protein was isolated and identified as Giardia eIF4E2. The other homologue, Giardia eIF4E1, bound only to the m(2,2,7)GpppN structure. Although neither homologue can rescue the function of yeast eIF4E, a knockdown of eIF4E2 mRNA in Giardia by a virus-based antisense ribozyme decreased translation, which was shown to use m(7)GpppN-capped mRNA as a template. Thus, eIF4E2 is likely the cap binding protein in a translation initiation complex. The same knockdown approach indicated that eIF4E1 is not required for translation in Giardia. Immunofluorescence assays showed wide distribution of both homologues in the cytoplasm. But eIF4E1 was also found concentrated and colocalized with the m(2,2,7)GpppN cap, 16S-like rRNA, and fibrillarin in the nucleolus-like structure in the nucleus. eIF4E1 depletion from Giardia did not affect mRNA splicing, but the protein was bound to Giardia small nuclear RNAs D and H known to have an m(2,2,7)GpppN cap, thus suggesting a novel function not yet observed among other eIF4Es in eukaryotes. PMID- 15879530 TI - Actively transcribing RNA polymerase II concentrates on spliced leader genes in the nucleus of Trypanosoma cruzi. AB - RNA polymerase II of trypanosomes, early diverging eukaryotes, transcribes long polycistronic messages, which are not capped but are processed by trans-splicing and polyadenylation to form mature mRNAs. The same RNA polymerase II also transcribes the genes coding for the spliced leader RNA, which are capped, exported to the cytoplasm, processed, and reimported into the nucleus before they are used as splicing donors to form mRNAs from pre-mRNA polycistronic transcripts. As pre-mRNA and spliced leader transcription events appear to be uncoupled, we studied how the RNA polymerase II is distributed in the nucleus of Trypanosoma cruzi. Using specific antibodies to the T. cruzi RNA polymerase II unique carboxy-terminal domain, we demonstrated that large amounts of the enzyme are found concentrated in a domain close to the parasite nucleolus and containing the spliced leader genes. The remaining RNA polymerase II is diffusely distributed in the nucleoplasm. The spliced leader-associated RNA polymerase II localization is dependent on the cell transcriptional state. It disperses when transcription is blocked by alpha-amanitin and actinomycin D. Tubulin genes are excluded from this domain, suggesting that it may exclusively be the transcriptional site of spliced leader genes. Trypomastigote forms of the parasite, which have reduced spliced leader transcription, show less RNA polymerase II labeling, and the spliced leader genes are more dispersed in the nucleoplasm. These results provide strong evidences that transcription of spliced leader RNAs occurs in a particular domain in the T. cruzi nucleus. PMID- 15879531 TI - Developmental variation in Rab11-dependent trafficking in Trypanosoma brucei. AB - In Trypanosoma brucei, endocytosis is developmentally regulated and is substantially more active in the mammalian infective stage, where it likely plays a role in immune evasion. The small GTPase TbRAB11 is highly expressed in the mammalian stage and mediates recycling of glycosylphosphatidylinositol-anchored proteins, including the variant surface glycoprotein (VSG) and the transferrin receptor, plus trafficking of internalized anti-VSG antibody and transferrin. No function has been assigned to TbRAB11 in the procyclic (insect) stage trypanosome. The importance of TbRAB11 to both bloodstream and procyclic form viability was assessed by RNA interference (RNAi). Suppression of TbRAB11 in the bloodstream form was rapidly lethal and led to cells with round morphology and an enlarged flagellar pocket. TbRAB11 RNAi was also lethal in procyclic forms, which also became rounded, but progression to cell death was significantly slower and the flagellar pocket remained normal. In bloodstream forms, silencing of TbRAB11 had no effect on exocytosis of newly synthesized VSG, fluid-phase endocytosis, or transferrin uptake, but export of internalized transferrin was inhibited. Lectin endocytosis assays revealed a block to postendosomal transport mediated by suppressing TbRAB11. By contrast, in procyclic forms, depletion of TbRAB11 blocks both fluid-phase endocytosis and internalization of surface proteins. In normal bloodstream forms, most VSG is recycled, but in procyclics, internalized surface proteins accumulated in the lysosome. These data demonstrate that TbRAB11 controls recycling and is essential in both life stages of T. brucei but that its primary role is subject to developmental variation. PMID- 15879539 TI - Prevalence and diagnosis of parasites of the stomach and small intestine in horses in south-west England. AB - Parasites were extracted from the stomach and small intestine of 118 horses at slaughter. The most abundant species was the tapeworm Anoplocephala perfoliata. Maximum likelihood analysis was used to investigate the relationship between the number of worms and their total weight, and the ability of an antibody-based elisa to diagnose the level of infection. The total weight of tapeworms increased towards a maximum as the number of worms increased, suggesting a population density-dependent constraint on the weight. The number of A perfoliata present could be predicted approximately from the results of the elisa. Although wide variation in elisa optical densities confound interpretation in individual animals, tests on groups of animals could provide a useful reflection of overall levels of infection. PMID- 15879532 TI - Class I histone deacetylase Thd1p affects nuclear integrity in Tetrahymena thermophila. AB - Class I histone deacetylases (HDACs) participate in the regulation of DNA templated processes such as transcription and replication. Members of this class can act locally at specific sites, or they can act more globally, contributing to a baseline acetylation state, both of which actions may be important for genome maintenance and organization. We previously identified a macronuclear-specific class I HDAC in Tetrahymena thermophila called Thd1p, which is expressed early in the development of the macronucleus when it initially becomes transcriptionally active. To test the idea that Thd1p is important for global chromatin integrity in an active macronucleus, Tetrahymena cells reduced in expression of Thd1p were generated. We observed phenotypes that indicated loss of chromatin integrity in the mutant cells, including DNA fragmentation and extrusion of chromatin from the macronucleus, variable macronuclear size and shape, enlarged nucleoli, and reduced phosphorylation of histone H1 from bulk chromatin. Macronuclei in mutant cells also contained more DNA. This observation suggests a role for Thd1p in the control of nuclear DNA content, a previously undescribed role for class I HDACs. Together, these phenotypes implicate Thd1p in the maintenance of macronuclear integrity in multiple ways, probably through site-specific changes in histone acetylation since no change in the acetylation levels of bulk histones was detected in mutant cells. PMID- 15879540 TI - Dysplasia of the caudal vertebral articular facets in four dogs: results of radiographic, myelographic and magnetic resonance imaging investigations. AB - Congenital anomalies of the vertebral column associated with aberrations of one of the primary vertebral ossification centres have been frequently described in the veterinary literature, but clinically significant abnormalities of secondary vertebral ossification centres, particularly involving the caudal articular processes, are much less frequently reported. This paper describes three dogs with aplasia and one dog with hypoplasia of the caudal vertebral articular processes. Thoracolumbar spinal cord compression and ataxia was evident in the three dogs with aplasia but no clinical signs were evident in the dog with hypoplasia. The radiographic appearance was similar in all four cases, with aplasia or hypoplasia of the caudal articular facets at one or more intervertebral joints in the thoracolumbar region. Bone proliferation was evident secondary to an associated degenerative joint disease. Compensatory hyperplasia of the adjacent cranial articular facets and ligamentum flavum protruded into the vertebral canal, resulting in a compressive myelopathy observed by myelography and magnetic resonance imaging. PMID- 15879541 TI - Efficacy of in-feed medication with tylosin for the treatment and control of Mycoplasma hyopneumoniae infections. AB - The efficacy of in-feed medication with tylosin for the treatment of enzootic pneumonia was examined in an experimental Mycoplasma hyopneumoniae infection model. One group of 10 conventional M. hyopneumoniae-free pigs was inoculated intratracheally with a highly virulent field isolate of M. hyopneumoniae; a second group of 10 pigs was inoculated in the same way and after 12 days was given tylosin at 100 mg/kg feed for 21 days; a third group of 10 pigs was inoculated with sterile culture medium, and these pigs were not given tylosin. The pigs were examined daily for clinical signs and each pig was given a respiratory disease score. Thirty-three days after they had been infected the pigs were euthanased, the lung lesions were quantified and samples of lung were processed for immunofluorescence testing for M. hyopneumoniae. The mean (sd) respiratory disease and lung lesion scores were significantly higher (P<0.05) in both the infected groups than in the uninfected group. Between 23 and 33 days after infection the mean respiratory disease score of the pigs treated with tylosin was 0.54 (0.22), significantly (P<0.05) lower than that of the infected pigs which were left untreated, 1.54 (0.46); similarly, their average lung lesion score, 1.72 (1.20), was significantly lower than that of the untreated pigs, 5.27 (3.85). PMID- 15879542 TI - Serological evidence of latency in cattle experimentally infected with elk herpesvirus. PMID- 15879543 TI - Gross and microscopic pathological findings in a sebaceous gland carcinoma of the perineum and vulva in a Friesian cow. PMID- 15879544 TI - Ileal diverticulum in a broiler chicken. PMID- 15879545 TI - Leptospiral seroepidemiology in a feline hospital population in Greece. PMID- 15879546 TI - Clinical evaluation of caudal epidural anaesthesia for the neutering of alpacas. PMID- 15879548 TI - Postweaning multisystemic wasting syndrome in pigs in Venezuela. PMID- 15879549 TI - Logos for the RCVS Practice Standards Scheme. PMID- 15879550 TI - Unwind and slow down: checkpoint activation by helicase and polymerase uncoupling. PMID- 15879551 TI - Neural guidance molecules regulate vascular remodeling and vessel navigation. AB - The development of the embryonic blood vascular and lymphatic systems requires the coordinated action of several transcription factors and growth factors that target endothelial and periendothelial cells. However, according to recent studies, the precise "wiring" of the vascular system does not occur without an ordered series of guidance decisions involving several molecules initially discovered for axons in the nervous system, including ephrins, netrins, slits, and semaphorins. Here, we summarize the new advances in our understanding of the roles of these axonal pathfinding molecules in vascular remodeling and vessel guidance, indicating that neuronal axons and vessel sprouts use common molecular mechanisms for navigation in the body. PMID- 15879552 TI - Default neural induction: neuralization of dissociated Xenopus cells is mediated by Ras/MAPK activation. AB - Xenopus embryonic ectodermal cells dissociated for three or more hours differentiate into neural tissue instead of adopting their normal epidermal fate. This default type of neural induction occurs in the absence of Spemann's organizer signals and is thought to be caused by the dilution of endogenous BMPs into the culture medium. Unexpectedly, we observed that BMP ligands continue to signal in dissociated cells. Instead, cell dissociation induces a sustained activation of the Ras/MAPK pathway, which causes the phosphorylation of Smad1 at sites that inhibit the activity of this transcription factor. It is this activation of Ras/MAPK that is required for neuralization in dissociated ectoderm. PMID- 15879553 TI - Brain lipid-binding protein is a direct target of Notch signaling in radial glial cells. AB - Radial glia function during CNS development both as neural progenitors and as a scaffolding supporting neuronal migration. To elucidate pathways involved in these functions, we mapped in vivo the promoter for Blbp, a radial glial gene. We show here that a binding site for the Notch effector CBF1 is essential for all Blbp transcription in radial glia, and that BLBP expression is significantly reduced in the forebrains of mice lacking the Notch1 and Notch3 receptors. These results identify Blbp as the first predominantly CNS-specific Notch target gene and suggest that it mediates some aspects of Notch signaling in radial glia. PMID- 15879554 TI - Lozenge directly activates argos and klumpfuss to regulate programmed cell death. AB - We show that reducing the activity of the Drosophila Runx protein Lozenge (Lz) during pupal development causes a decrease in cell death in the eye. We identified Lz-binding sites in introns of argos (aos) and klumpfuss (klu) and demonstrate that these genes are directly activated targets of Lz. Loss of either aos or klu reduces cell death, suggesting that Lz promotes apoptosis at least in part by regulating aos and klu. These results provide novel insights into the control of programmed cell death (PCD) by Lz during Drosophila eye development. PMID- 15879555 TI - Specific signals at the 3' end of the DHFR gene define one boundary of the downstream origin of replication. AB - The Chinese hamster dihydrofolate reductase (DHFR) origin of replication consists of a 55-kb zone of potential initiation sites lying between the convergently transcribed DHFR and 2BE2121 genes. Two subregions within this zone (ori-beta/ori beta' and ori-gamma) are preferred. In the DHFR-deficient variant, DR8, which has deleted a 14-kb sequence straddling the 3' end of the DHFR gene, early-firing origin activity in the downstream ori-beta/ori-beta' and ori-gamma regions is completely suppressed. We show that the critical deleted sequences reside within a 168-bp segment encompassing the intron 5/exon 6 boundary, exon 6, 54 bp of the 3' untranslated region (UTR), but not the three natural polyA sites. In wild-type cells, this sequence efficiently arrests transcription in a region a few kilobases downstream, which coincides with the 5' boundary of the replication initiation zone. In DR8, DHFR-specific transcripts efficiently use an alternative sixth exon (6c) and polyA signals near the middle of the former intergenic region to process primary transcripts. However, transcription proceeds to a position almost 35 kb downstream from these signals, and replication initiation can only be detected beyond this point. When the wild-type 168-bp 3' element is inserted into DR8 at the same position as alternative exon 6c, transcription is arrested efficiently and initiations occur almost immediately downstream. Thus, the normal 3' end of the DHFR gene constitutes a boundary element not only for the gene but also for the local origin of replication. PMID- 15879556 TI - Polarity of the mouse embryo is established at blastocyst and is not prepatterned. AB - Polarity formation in mammalian preimplantation embryos has long been a subject of controversy. Mammalian embryos are highly regulative, which has led to the conclusion that polarity specification does not exist until the blastocyst stage; however, some recent reports have now suggested polarity predetermination in the egg. Our recent time-lapse recordings have demonstrated that the first cleavage plane is not predetermined in the mouse egg. Here we show that, in contrast to previous claims, two-cell blastomeres do not differ and their precise future contribution to the inner cell mass and/or the trophectoderm cannot be anticipated. Thus, all evidence so far strongly suggests the absence of predetermined axes in the mouse egg. We observe that the ellipsoidal zona pellucida exerts mechanical pressure and space constraints as the coalescing multiple cavities are restricted to one end of the long axis of the blastocyst. We propose that these mechanical cues, in conjunction with the epithelial seal in the outer cell layer, lead to specification of the embryonic-abembryonic axis, thus establishing first polarity in the mouse embryo. PMID- 15879557 TI - Overexpression of fasciculation and elongation protein zeta-1 (FEZ1) induces a post-entry block to retroviruses in cultured cells. AB - Two mutant Rat2 fibroblast cell lines, R3-2 and R4-7, have been previously isolated by a selection for retrovirus resistance. We have now further analyzed the basis of the block to retroviral infection in the R3-2 line. Using Affymetrix GeneChip analysis, several genes were identified as differentially expressed in the mutant R3-2 line compared with the wild-type cells. One of the candidate gene products, FEZ1 (fasciculation and elongation protein zeta-1), a protein kinase C (PKC)zeta-interacting protein homologous to the Caenorhabditis elegans synaptic transport protein UNC-76, was found to be up-regulated >30-fold in the resistant R3-2 line. FEZ1 overexpression in Rat2 cells conferred a potent resistance to infection by genetically marked retroviruses, and the degree of retroviral resistance in both Rat2 fibroblasts and 293T cells tightly correlated with the expression level of FEZ1 transcripts. FEZ1-overexpressing Rat2 cells showed a similar phenotype to that of the mutant R3-2 line: Infection resulted in normal viral DNA synthesis but a reduction in the formation of circular DNA, indicating a block after reverse transcription but before nuclear entry. Partial knockdown of FEZ1 expression in R3-2 by RNA interference (RNAi) significantly reduced the resistance of this line to infection. Thus, our data suggest that FEZ1 overexpression is sufficient to explain the resistant phenotype of R3-2 cells and identify FEZ1 as a new gene capable of causing retrovirus resistance. PMID- 15879558 TI - The glucocorticoid receptor blocks P-TEFb recruitment by NFkappaB to effect promoter-specific transcriptional repression. AB - To investigate the determinants of promoter-specific gene regulation by the glucocorticoid receptor (GR), we compared the composition and function of regulatory complexes at two NFkappaB-responsive genes that are differentially regulated by GR. Transcription of the IL-8 and IkappaBalpha genes is stimulated by TNFalpha in A549 cells, but GR selectively represses IL-8 mRNA synthesis by inhibiting Ser2 phosphorylation of the RNA polymerase II (pol II) C-terminal domain (CTD). The proximal kappaB elements at these genes differ in sequence by a single base pair, and both recruited RelA and p50. Surprisingly, GR was recruited to both of these elements, despite the fact that GR failed to repress the IkappaBalpha promoter. Rather, the regulatory complexes formed at IL-8 and IkappaBalpha were distinguished by differential recruitment of the Ser2 CTD kinase, P-TEFb. Disruption of P-TEFb function by the Cdk-inhibitor, DRB, or by small interfering RNA selectively blocked TNFalpha stimulation of IL-8 mRNA production. GR competed with P-TEFb recruitment to the IL-8 promoter. Strikingly, IL-8 mRNA synthesis was repressed by GR at a post-initiation step, demonstrating that promoter proximal regulatory sequences assemble complexes that impact early and late stages of mRNA synthesis. Thus, GR accomplishes selective repression by targeting promoter-specific components of NFkappaB regulatory complexes. PMID- 15879559 TI - Localization of the microtubule end binding protein EB1 reveals alternative pathways of spindle development in Arabidopsis suspension cells. AB - In a previous study on Arabidopsis thaliana suspension cells transiently infected with the microtubule end binding protein AtEB1a-green fluorescent protein (GFP), we reported that interphase microtubules grow from multiple sites dispersed over the cortex, with plus ends forming the characteristic comet-like pattern. In this study, AtEB1a-GFP was used to study the transitions of microtubule arrays throughout the division cycle of cells lacking a defined centrosome. During division, the dispersed origin of microtubules was replaced by a more focused pattern with the plus end comets growing away from sites associated with the nuclear periphery. The mitotic spindle then evolved in two quite distinct ways depending on the presence or absence of the preprophase band (PPB): the cells displaying outside-in as well as inside-out mitotic pathways. In those cells possessing a PPB, the fusion protein labeled material at the nuclear periphery that segregated into two polar caps, perpendicular to the PPB, before nuclear envelope breakdown (NEBD). These polar caps then marked the spindle poles upon NEBD. However, in the population of cells without PPBs, there was no prepolarization of material at the nuclear envelope before NEBD, and the bipolar spindle only emerged clearly after NEBD. Such cells had variable spindle orientations and enhanced phragmoplast mobility, suggesting that the PPB is involved in a polarization event that promotes early spindle pole morphogenesis and subsequent positional stability during division. Astral-like microtubules are not usually prominent in plant cells, but they are clearly seen in these Arabidopsis cells, and we hypothesize that they may be involved in orienting the division plane, particularly where the plane is not determined before division. PMID- 15879560 TI - Dual role for tomato heat shock protein 21: protecting photosystem II from oxidative stress and promoting color changes during fruit maturation. AB - The tomato (Lycopersicon esculentum) chloroplast small heat shock protein (sHSP), HSP21, is induced by heat treatment in leaves, but also under normal growth conditions in developing fruits during the transition of chloroplasts to chromoplasts. We used transgenic tomato plants constitutively expressing HSP21 to study the role of the protein under stress conditions and during fruit maturation. Although we did not find any effect for the transgene on photosystem II (PSII) thermotolerance, our results show that the protein protects PSII from temperature-dependent oxidative stress. In addition, we found direct evidence of the protein's role in fruit reddening and the conversion of chloroplasts to chromoplasts. When plants were grown under normal growth temperature, transgenic fruits accumulated carotenoids earlier than controls. Furthermore, when detached mature green fruits were stored for 2 weeks at 2 degrees C and then transferred to room temperature, the natural accumulation of carotenoids was blocked. In a previous study, we showed that preheat treatment, which induces HSP21, allowed fruit color change at room temperature, after a cold treatment. Here, we show that mature green transgenic fruits constitutively expressing HSP21 do not require the heat treatment to maintain the ability to accumulate carotenoids after cold storage. This study demonstrates that a sHSP plays a role in plant development under normal growth conditions, in addition to its protective effect under stress conditions. PMID- 15879562 TI - Health care workers' expectations and empathy toward patients in abusive relationships. AB - PURPOSE: To understand attitudes that may affect health care workers' ability to provide appropriate long-term care for patients who stay with abusive partners. METHODS: We surveyed 278 health care workers in 31 primary care practices before their participation in an educational intervention. RESULTS: More than half of participants (51% to 60%) found it easy to empathize with a patient who decided to remain in an abusive relationship if the patient was described as poor or disabled, but few (25% to 39%) could empathize if the patient was described as educated or financially secure. A majority (57% to 59%) agreed with a statement meant to assess unrealistic expectations. ("A provider's responsibility includes making sure a patient gets to a shelter right away if he or she discloses abuse.") Participants who agreed with this statement had more difficulty empathizing with patients who decided to stay with an abusive partner (P = .0045). CONCLUSIONS: Training efforts must focus on screening and on helping providers develop more realistic expectations about the complex nature of leaving an abusive relationship. Health care workers need a better understanding of the barriers patients face and why patients may choose to remain in abusive relationships, even in the absence of economic or health limitations. PMID- 15879561 TI - Class 1 reversibly glycosylated polypeptides are plasmodesmal-associated proteins delivered to plasmodesmata via the golgi apparatus. AB - SE-WAP41, a salt-extractable 41-kD wall-associated protein that is associated with walls of etiolated maize (Zea mays) seedlings and is recognized by an antiserum previously reported to label plasmodesmata and the Golgi, was cloned, sequenced, and found to be a class 1 reversibly glycosylated polypeptide ((C1)RGP). Protein gel blot analysis of cell fractions with an antiserum against recombinant SE-WAP41 showed it to be enriched in the wall fraction. RNA gel blot analysis along the mesocotyl developmental axis and during deetiolation demonstrates that high SE-WAP41 transcript levels correlate spatially and temporally with primary and secondary plasmodesmata (Pd) formation. All four of the Arabidopsis thaliana (C1)RGP proteins, when fused to green fluorescent protein (GFP) and transiently expressed in tobacco (Nicotiana tabacum) epidermal cells, display fluorescence patterns indicating they are Golgi- and plasmodesmal associated proteins. Localization to the Golgi apparatus was verified by colocalization of transiently expressed AtRGP2 fused to cyan fluorescence protein together with a known Golgi marker, Golgi Nucleotide Sugar Transporter 1 fused to yellow fluorescent protein (GONST1:YFP). In transgenic tobacco, AtRGP2:GFP fluorescence is punctate, is present only in contact walls between cells, and colocalizes with aniline blue-stained callose present around Pd. In plasmolyzed cells, AtRGP2:GFP remains wall embedded, whereas GONST1:YFP cannot be found embedded in cell walls. This result implies that the targeting to Pd is not due to a default pathway for Golgi-localized fusion proteins but is specific to (C1)RGPs. Treatment with the Golgi disrupting drug Brefeldin A inhibits Pd labeling by AtRGP2:GFP. Integrating these data, we conclude that (C1)RGPs are plasmodesmal-associated proteins delivered to plasmodesmata via the Golgi apparatus. PMID- 15879563 TI - Connections to primary medical care after psychiatric crisis. AB - BACKGROUND: Patients presenting with a psychiatric emergency face a unique set of challenges in connecting to primary care. OBJECTIVES: We tested the hypothesis that, in contrast to usual care, case management will result in higher rates of connection to primary care. We examined variables affecting primary care entry, including insurance status, hospital admission, and concurrent linkages to mental health care. RESEARCH DESIGN/METHODS: This article reports on a preliminary outcome of an ongoing randomized controlled trial conducted with 101 patients presenting in an urban psychiatric setting. Patients were randomized to a case management team or to usual care. The need for medical care was assessed by documenting medical comorbidity. RESULTS: Average age of the sample was 37.5; 65% were male, and 78% had low income; 37% were African American and 9% were Hispanic. Within 3 months of study enrollment, 57% of the intervention group was successfully linked to primary care compared with 16% of the usual care group, a difference that was statistically significant (P < .001). Associated positive predictors for linkage to primary care included mental health care visits and success in obtaining health insurance. Inpatient hospital stay at the time of psychiatric crisis was negatively associated with later attendance at primary care. CONCLUSIONS: Case management intervention was effective in establishing linkage to primary care within 3 months. Ongoing work will evaluate primary care retention and physical and mental health outcomes. PMID- 15879564 TI - The sexual concerns of african american, asian american, and white women seeking routine gynecological care. AB - PURPOSE: Because of limited knowledge regarding the sexual health of minority women, we described the types and frequency of sexual concerns among African American, Asian American, and white women. METHODS: The sexual concerns of eligible women who received their health care at a large military health care facility in Washington State were determined from December 1992 through January 1993 through the use of a questionnaire on women's sexual health that was developed by one of the authors. RESULTS: A high and similar prevalence of sexual concerns was reported among these women. The prevalence of certain sexual concerns among white, African American, and Asian American women differed significantly. A history of sexual coercion or physical/emotional abuse was strongly associated with a high prevalence of sexual concerns for each racial and ethnic group. CONCLUSION: Although women of different racial/ethnic backgrounds reported a high and similar prevalence of sexual concerns, there were significant differences in the prevalence of certain sexual concerns that may have differential health importance. Health-care providers should exercise caution when generalizing the sexual concerns of women from different racial/ethnic backgrounds. PMID- 15879565 TI - Enhancing doctor-patient communication using email: a pilot study. AB - BACKGROUND: The doctor-patient relationship has been eroded by many factors. Would e-mail enhance communication and address some of the barriers inherent to our medical practices? METHODS: Of our study population, 4 physicians offered e mail communication to participating patients and 4 did not. Both patients and physicians completed questionnaires regarding satisfaction, perceived quality, convenience, and promptness of the communication. RESULTS: Patient satisfaction significantly increased in the e-mail group compared with the control group in the areas of convenience (P < .0001) and the amount of time spent contacting their physician (P < .0001). Physician satisfaction in the e-mail group increased regarding convenience, amount of time spent on messages, and volume of messages. The response time was longer with e-mail. When asked if patients should be able to e-mail their physicians, most patients in the e-mail group and all but 2 of the physicians in the non-e-mail group responded "yes." CONCLUSION: E-mail communication was found to be a more convenient form of communication. Satisfaction by both patients and physicians improved in the e-mail group. The volume of messages and the time spent answering messages for the e-mail group physicians was not increased. E-mail has the potential to improve the doctor patient relationship as a result of better communication. PMID- 15879566 TI - Diastolic heart failure: a review and primary care perspective. AB - Previously the subject of much debate, there is now consensus that diastolic heart failure (DHF) represents a distinct form of heart failure. Epidemiologic data indicate that DHF is common. Indeed, there is evidence that, among elderly persons, DHF is more common than systolic heart failure (SHF). Like SHF, DHF is associated with significant morbidity, mortality, and cost; however, few clinical trials focusing on isolated DHF have been completed. Much of the treatment of DHF is based on current concepts of the pathophysiology of DHF, small clinical studies, and experience gained from treating patients with SHF. The diagnosis of DHF is clinical; data supporting the establishment of a diagnosis of DHF are limited. Differences exist in prognosis and treatment between diastolic and systolic heart failure. This article reviews diastolic heart failure with emphasis on evidence-based management, aimed at primary care physicians who routinely provide care to patients with DHF. PMID- 15879567 TI - Basal insulin therapy in type 2 diabetes. AB - Patients with type 2 diabetes mellitus are usually treated initially with oral antidiabetic agents, but as the disease progresses, most patients eventually require insulin to maintain glucose control. Optimal insulin therapy should mimic the normal physiologic secretion of insulin and minimize the risk of hypoglycemia. This article discusses the role of insulin therapy in patients with type 2 diabetes, emphasizing long-acting insulin agents designed to approximate physiologic basal insulin secretion and provide control over fasting plasma glucose. Clinical trials of recently developed long-acting insulins are reviewed herein, with emphasis on studies that combined basal insulin with oral agents or with short-acting insulins in a basal-bolus approach. The normal physiologic pattern of insulin secretion by pancreatic beta cells consists of a sustained basal insulin level throughout the day, superimposed after meals by relatively large bursts of insulin that slowly decay over 2 to 3 hours (bolus insulin). Basal support with long-acting insulin is a key component of basal-bolus therapy for patients with diabetes who require insulin with or without the addition of oral agents. Newer long-acting agents such as insulin glargine provide a steadier and more reliable level of basal insulin coverage and may have significant advantages over traditional long-acting insulins as part of a basal-bolus treatment strategy. PMID- 15879568 TI - Dietary calcium intake and obesity. AB - Obesity is increasing in the United States in epidemic proportions. Epidemiologic data suggest that people with high calcium intake have a lower prevalence of overweight, obesity, and insulin resistance syndrome. Studies in transgenic mice have demonstrated that calcium influences adipocyte metabolism. High calcium intake depresses levels of parathyroid hormone and 1,25-hydroxy vitamin D. These decreased hormone levels cause decreases in intracellular calcium, thereby inhibiting lipogenesis and stimulating lipolysis. High dietary calcium intakes also increases excretion of fecal fat and may increase core body temperature. Calcium from dairy products seems to have more of an impact than calcium from dietary supplements. Primary care providers should include recommendations about adequate calcium intake in standard dietary counseling about weight management. PMID- 15879569 TI - Patients with a family history of cancer: identification and management. AB - A family history of certain malignancies, especially breast, ovarian, colorectal, and prostate cancers, can place persons at increased risk of developing these cancers. By constructing a pedigree that includes 3 generations, family physicians can identify patients at increased risk because of family cancer history. Persons at increased cancer risk because of family history warrant a surveillance strategy for early detection. Genetic professionals represent an important resource in assessing genetic risk and possible testing. Persons identified as being at increased risk of various cancers based on their family history should understand the surveillance plan that is recommended and the importance of maintaining a healthy lifestyle and remaining up to date on other cancer screening tests. PMID- 15879570 TI - Health care interactions with deaf culture. PMID- 15879571 TI - An enhanced obstetrics track for a family practice residency program: results from the first 6 years. AB - BACKGROUND: Advanced training in obstetrics for family physicians occurs through a variety of methods. The program described has developed an obstetrics track for family practice residents. METHODS: Five residents have completed the 4-year residency program with enhanced obstetric training developed, and the results, in terms of procedural experience and examination scores, have been reviewed. RESULTS: These 5 family physicians performed a similar number of obstetric procedures compared with their Obstetrics and Gynecology resident counterparts, and they performed as well as their family medicine resident counterparts on national in-service examinations. CONCLUSIONS: A 4-year enhanced obstetrics track is an effective means of improving the training of family medicine residents in obstetric procedures while maintaining the other fundamental training and residency review committee requirements for family medicine residents. PMID- 15879572 TI - Development of species-specific rDNA probes for Giardia by multiple fluorescent in situ hybridization combined with immunocytochemical identification of cyst wall antigens. AB - In this study, we describe the development of fluorescent oligonucleotide probes to variable regions in the small subunit of 16S rRNA in three distinct Giardia species. Sense and antisense probes (17-22 mer) to variable regions 1, 3, and 8 were labeled with digoxygenin or selected fluorochomes (FluorX, Cy3, or Cy5). Optimal results were obtained with fluorochome-labeled oligonucleotides for detection of rRNA in Giardia cysts. Specificity of fluorescent in situ hybridization (FISH) was shown using RNase digestion and high stringency to diminish the hybridization signal, and oligonucleotide probes for rRNA in Giardia lamblia, Giardia muris, and Giardia ardeae were shown to specifically stain rRNA only within cysts or trophozoites of those species. The fluorescent oligonucleotide specific for rRNA in human isolates of Giardia was positive for ten different strains. A method for simultaneous FISH detection of cysts using fluorescent antibody (genotype marker) and two oligonucleotide probes (species marker) permitted visualization of G. lamblia and G. muris cysts in the same preparation. Testing of an environmental water sample revealed the presence of FISH-positive G. lamblia cysts with a specific rDNA probe for rRNA, while negative cysts were presumed to be of animal or bird origin. PMID- 15879573 TI - Targeted deletion of the SPARC gene accelerates disc degeneration in the aging mouse. AB - SPARC (secreted protein, acidic, and rich in cysteine) is a matricellular protein that is present in the intervertebral disc; in man, levels of SPARC decrease with aging and degeneration. In this study, we asked whether targeted deletion of SPARC in the mouse influenced disc morphology. SPARC-null and wild-type (WT) mice were studied at 0.3-21 months of age. Radiologic examination of spines from 2 month-old SPARC-null mice revealed wedging, endplate calcification, and sclerosis, features absent in age-matched WT spines. Discs from 3-month-old SPARC null mice had a greater number of annulus cells than those of WT animals (1884.6 +/- 397.9 [mean +/- SD] vs 1500.2 +/- 188.2, p=0.031). By 19 months discs from SPARC-null mice contained fewer cells than WT counterparts (1383.6 +/- 363.3 vs 1466.8 +/- 148.0, p=0.033). Histology of midsagittal spines showed herniations of lower lumbar discs of SPARC-null mice ages 14-19 months; in contrast, no herniations were seen in WT age-matched animals. Ultrastructural studies showed uniform collagen fibril diameters in the WT annulus, whereas in SPARC-null disc fibrils were of variable size with irregular margins. Consistent with the connective tissue deficits observed in other tissues of SPARC-null mice, our findings support a fundamental role for SPARC in the production, assembly, or maintenance of the disc extracellular matrix. PMID- 15879574 TI - Overexpression of eCLCA1 in small airways of horses with recurrent airway obstruction. AB - The human hCLCA1 and murine mCLCA3 (chloride channels, calcium-activated) have recently been identified as promising therapeutic targets in asthma. Recurrent airway obstruction in horses is an important animal model of human asthma. Here, we have cloned and characterized the first equine CLCA family member, eCLCA1. The 913 amino acids eCLCA1 polypeptide forms a 120-kDa transmembrane glycoprotein that is processed to an 80-kDa protein in vivo. Three single nucleotide polymorphisms were detected in the eCLCA1 coding region in 14 horses, resulting in two amino acid changes (485H/R and 490V/L). However, no functional differences were recorded between the channel properties of the two variants in transfected HEK293 cells. The eCLCA1 protein was detected immunohistochemically in mucin producing cells in the respiratory and intestinal tracts, cutaneous sweat glands, and renal mucous glands. Strong overexpression of eCLCA1 was observed in the airways of horses with recurrent airway obstruction using Northern blot hybridization, Western blotting, immunohistochemistry, and real-time quantitative RT-PCR. The results suggest that spontaneous or experimental recurrent airway obstruction in horses may serve as a model to study the role of CLCA homologs in chronic airway disease with overproduction of mucins. PMID- 15879575 TI - Immunocytochemical localization of atrial natriuretic peptide, vessel dilator, long-acting natriuretic peptide, and kaliuretic peptide in human pancreatic adenocarcinomas. AB - We recently found that four peptide hormones synthesized by the same gene completely inhibit the growth of human pancreatic adenocarcinomas in athymic mice. The present immunocytochemical investigation was designed to determine where in the adenocarcinomas these peptide hormones localize. Atrial natriuretic peptide, vessel dilator, long-acting natriuretic peptide, and kaliuretic peptide localized to the cytoplasm and nucleus of the human pancreatic adenocarcinomas, which is consistent with their ability to decrease DNA synthesis in the nucleus of this cancer. In this first investigation of where these peptide hormones with anticancer effects localize in any cancer, these peptide hormones also localized to the endothelium of capillaries and fibroblasts within these cancers. This is the first demonstration of growth-inhibiting peptide hormones localizing to the nucleus, where they inhibit DNA synthesis and may interact with growth-promoting hormones that localize there as the etiology of their ability to inhibit the growth of adenocarcinomas both in vitro and in vivo. PMID- 15879576 TI - Profiles of PrKX expression in developmental mouse embryo and human tissues. AB - Protein kinase X (PrKX), karyotypically located on the human X chromosome, is a type I cAMP-dependent protein kinase. Although a specific role for PrKX has not yet been defined, PrKX gene expression in mouse and human tissues has been profiled only by in situ hybridization and Northern blot analyses and not by protein expression. To determine more precisely the PrKX protein levels, we developed specific anti-PrKX antibodies and examined gestationally staged mouse embryo sections by immunohistochemistry. These results showed that PrKX is ubiquitously distributed and highly expressed in murine central nervous system and heart tissues in early developmental stages and in most organs at later stages but was not detected in either connective tissues or bone. Using Western blots to detect PrKX, total protein extracts from eight different adult or fetal human tissues including brain, heart, kidney, liver, lung, pancreas, spleen, and thymus were analyzed. Although PrKX protein was present in each of the tissues tested, the protein levels varied depending on tissue type and developmental stage. Very low protein levels were found in heart tissues from a 5-month-old fetus and from an adult, whereas PrKX proteins were more abundant in fetal brain, kidney, and liver tissues compared with adult samples of the same tissue type. PMID- 15879577 TI - Parathyroid hormone-related peptide (hPTHrP) and parathyroid hormone-related peptide receptor type 1 (PTHR1) expression in human thymus. AB - Parathyroid hormone-related peptide (hPTHrP) is expressed in human tissues and regulates cellular proliferation, differentiation, and apoptosis by an autocrine/paracrine loop. In rodent thymus, both parathormone and parathyroid hormone-related peptide (PTHrP) are expressed by thymic epithelial cells (TECs). The present study demonstrated by RT-PCR and immunohistochemistry that hPTHrP and parathyroid hormone-related peptide receptor type 1 (PTHR1) were expressed in human thymus at both RNA and protein levels. hPTHrP was expressed mainly in the thymic medulla by epithelial (cytokeratin-positive), mature dendritic (CD40+/86+) and plasmacytoid interleukin (IL)-3Ralpha1 cells. This protein was also present in some cells forming Hassall's bodies and a few subcapsular and cortical TECs. PTHR1 was expressed by scattered subcapsular and cortical TECs and by rare TECs in the medulla. Thymocytes did not express either hPTHrP or PTHR1. Primary cultures of human TECs revealed the presence of both hPTHrP and PTHR1 mRNAs, confirming the capacity of TECs to synthesize both peptides. Moreover, synthetic (1-39) hPTHrP peptide administered on cultured TECs induced the expression of IL 6 mRNA, suggesting that hPTHrP can regulate thymic functions by inducing in TECs the expression of IL-6, which is involved in the development and maturation of thymocytes. PMID- 15879578 TI - Ultrastructural studies and Na+,K+-ATPase immunolocalization in the antennal urinary glands of the lobster Homarus gammarus (Crustacea, Decapoda). AB - Unlike in crustacean freshwater species, the structure and ultrastructure of the excretory antennal gland is poorly documented in marine species. The general organization and ultrastructure of the cells and the localization of Na(+),K(+) ATPase were examined in the antennal gland of the adult lobster Homarus gammarus. Each gland is composed of a centrally located coelomosac surrounded ventrally by a labyrinth divided into two parts (I and II) and dorsally by a voluminous bladder. There is no differentiated nephridal tubule between them. The labyrinth and bladder cells have in common a number of ultrastructural cytological features, including basal membrane infoldings associated with mitochondria, apical microvilli, and cytoplasmic extrusions, and a cytoplasm packed with numerous vacuoles, vesicles, lysosome-like bodies, and swollen mitochondria. Each type of cell also presents distinctive characters. Na(+),K(+)-ATPase was detected through immunofluorescence in the basal part of the cells of the labyrinth and in the bladder cells with an increasing immunostaining from labyrinth I to the bladder. No immunoreactivity was detected in the coelomosac. The cells of the labyrinth and of the bladder present morphological and enzymatic features of ionocytes. The antennal glands of the lobster thus possess active ion exchanges capabilities. PMID- 15879579 TI - Depth-varying density and organization of chondrocytes in immature and mature bovine articular cartilage assessed by 3d imaging and analysis. AB - Articular cartilage is a heterogeneous tissue, with cell density and organization varying with depth from the surface. The objectives of the present study were to establish a method for localizing individual cells in three-dimensional (3D) images of cartilage and quantifying depth-associated variation in cellularity and cell organization at different stages of growth. Accuracy of nucleus localization was high, with 99% sensitivity relative to manual localization. Cellularity (million cells per cm3) decreased from 290, 310, and 150 near the articular surface in fetal, calf, and adult samples, respectively, to 120, 110, and 50 at a depth of 1.0 mm. The distance/angle to the nearest neighboring cell was 7.9 microm/31 degrees , 7.1 microm/31 degrees , and 9.1 microm/31 degrees for cells at the articular surface of fetal, calf, and adult samples, respectively, and increased/decreased to 11.6 microm/31 degrees , 12.0 microm/30 degrees , and 19.2 microm/25 degrees at a depth of 0.7 mm. The methodologies described here may be useful for analyzing the 3D cellular organization of cartilage during growth, maturation, aging, degeneration, and regeneration. PMID- 15879580 TI - Expression of components of the renin-angiotensin system in autosomal recessive polycystic kidney disease. AB - Hypertension is a common complication in children with autosomal recessive polycystic kidney disease (ARPKD) who have survived the neonatal period. No information is available regarding the mechanism of hypertension in this condition. The renin-angiotensin system (RAS) is thought to play a role in hypertension associated with the more common autosomal dominant polycystic kidney disease (ADPKD). Occasional reports have documented increased activity of the intrarenal RAS in ADPKD, with ectopic renin expression within cysts and dilated tubules. Because of similarities between ARPKD and ADPKD, we hypothesized that increased intrarenal RAS activity might also be found in ARPKD. We performed immunohistochemical studies on kidney tissues from two infants with ARPKD and two control kidneys. The cystic dilated tubules showed staining with the peanut lectin arachis hypogaea, a marker of distal tubules and collecting ducts, but not with lotus tetragonolobus, a marker of proximal tubules. Strong renin staining was seen in many cysts and tubules of ARPKD kidneys, but only in the afferent arterioles of the normal control kidneys. Angiotensinogen staining was also observed in some cysts and in proximal tubules. Staining for angiotensin converting enzyme, angiotensin II type 1 receptor, and angiotensin II peptide was present in many cystic dilated tubules. These immunohistochemical studies document for the first time ectopic expression of components of the RAS in cystic dilated tubules of ARPKD and suggest that overactivity of RAS could result in increased intrarenal angiotensin II production, which may contribute to the development of hypertension in ARPKD. PMID- 15879581 TI - Sleep and aging. PMID- 15879582 TI - Aging and the role of the HPA axis and rhythm in sleep and memory-consolidation. AB - Changes in the hypothalamo-pituitary-adrenal (HPA) axis and its rhythm with aging have interesting implications for sleep. Herein, the authors review sleep and HPA changes associated with normal aging and point out the similarities in how they change over time. The authors also discuss the effects of sleep on declarative memory consolidation, in particular. This focused review suggests that some of the declarative memory dysfunction with normal aging, and possibly procedural memory dysfunction, may be partially reversible by instituting methods to augment slow-wave sleep (SWS). Also, agents that decrease nocturnal corticotropin releasing hormone and the cortisol nadir and enhance SWS may offer potential ways to manipulate the HPA axis/rhythm and improve sleep and memory. In this regard, the authors propose that drugs that act directly on the HPA axis (e.g., mineralocorticoid agonists) may be potentially quite useful for improving both sleep and declarative memory consolidation during sleep. PMID- 15879583 TI - Accelerating symptom-reduction in late-life depression: a double-blind, randomized, placebo-controlled trial of sleep deprivation. AB - OBJECTIVE: Authors tested the hypothesis that one night of total sleep deprivation (TSD) would accelerate antidepressant response to paroxetine, as compared with TSD+placebo (PBO) and paroxetine-alone, in late-life major depression. METHODS: Eighty elderly outpatients with current episodes of non psychotic, non-bipolar major depression were randomly assigned to one of three treatment conditions: TSD+paroxetine (N = 27), TSD + PBO (N = 27), and paroxetine only (N = 26). Primary outcome was percentage of subjects in each condition who demonstrated early response (Hamilton Rating Scale for Depression scores [Ham-D: 17-item] of < or = 10) or remission (score of < or = 7) on Day 14. RESULTS: Response rates after 14 days were 22% in subjects randomly assigned to the TSD + paroxetine condition, 41% in TSD + PBO, and 46% in paroxetine alone. Remission rates after 14 days were 11% in TSD+paroxetine, 22% in TSD + PBO, and 38% in paroxetine. After adjusting for baseline depression severity, there were no statistically significant differences in response or remission rates. CONCLUSION: Contrary to the study hypothesis, one night of total sleep deprivation did not accelerate onset of antidepressant response to paroxetine pharmacotherapy of late life depression. The data suggest, rather, that the two interventions might have counteracted each other. PMID- 15879584 TI - Disturbance of endogenous circadian rhythm in aging and Alzheimer disease. AB - OBJECTIVE: Normal sleep-wake regulation is dependent upon an oscillatory circadian rhythm promoting alertness and sleep at appropriate times of day. Circadian rhythms have been noted to be disturbed as a consequence of both normal aging and age-associated pathologies like Alzheimer disease (AD). However, the relationship between the consequences of normal versus pathological aging upon circadian regulation remains unclear. The authors evaluated the similarities and differences between the consequences of aging and AD on endogenous circadian rhythm. METHODS: Authors measured locomotor activity and, with a constant routine protocol, core body temperature, examining differences and similarities in circadian disturbances in groups of normal elderly and patients with probable AD (pAD), as compared with a comparison group of young, normal volunteers, measuring endogenous circadian amplitude (ECA) and endogenous circadian phase (ECP) of core body temperature, and made parametric and nonparametric assessments of locomotor activity rhythms. RESULTS: The ECP of core body temperature was delayed in patients with pAD versus both normal young and normal elderly subjects, whereas the ECA was reduced both in normal elderly and pAD subjects compared with a comparison group of young subjects. There was also disassociation of the activity and core body temperature rhythms in both age groups versus the young subjects. CONCLUSIONS: Authors observed changes in endogenous circadian rhythm in pAD that were consonant with those seen in normal aging (amplitude reduction; loss of phase coordination) and also observed changes that were apparently discrete from those seen in normal aging (phase delay). PMID- 15879585 TI - Passive body heating ameliorates sleep disturbances in patients with vascular dementia without circadian phase-shifting. AB - OBJECTIVE: This study investigated the sleep-promoting, thermoregulatory, and circadian phase-shifting actions of passive body heating (PBH) in elderly insomniac patients (IPs) with mild-to-moderate vascular dementia. METHODS: Thirteen elderly IPs with vascular dementia (mean age 76.9 years; male/female ratio 2/11) were subjected to a PBH trial session. This session comprised a 3-day baseline period, 2-day PBH period, and 1-day post-PBH period. In the PBH period, the subjects received PBH (immersion in hot water about 40.0 degrees C to mid thorax level) for 30 minutes beginning 2 hours before bedtime. Sleep-waking, estimated by actigraph, core body temperature (cBT), and heart rate variability were continuously monitored. Dim-light melatonin-onset time (DLMO) was determined in the baseline and post-PBH periods. RESULTS: PBH significantly improved subjects' sleep quality; sleep latency decreased; sleep efficiency increased; and wake time after sleep onset decreased. These trends were more prominent in the latter half of the sleep time. PBH induced a rapid cBT elevation of approximately 0.80 degrees C, on average, followed by enhanced heat loss (DeltacBT: difference in cBT between just after the PBH and bedtime), lasting 1.5 hours before sleep. There was a significantly positive correlation between DeltacBT and sleep latency. PBH induced no significant phase shift in DLMO. Heart-rate variability data showed that PBH induced parasympathomimetic action during sleep time in the subjects. CONCLUSION: PBH may have a sleep-promoting effect by intervening in the thermoregulatory and autonomic systems in elderly IPs with vascular dementia. PMID- 15879586 TI - Influence of aging on the improvement of subjective sleep quality by atypical antipsychotic drugs in patients with schizophrenia: comparison of middle-aged and older adults. AB - OBJECTIVE: The authors investigated the influence of aging on the improvement of subjective sleep quality by atypical antipsychotic drugs in patients with schizophrenia. METHODS: Subjects were 86 inpatients (mean age: 61.4 years) who had been receiving treatment with conventional antipsychotic drugs and who met DSM-IV criteria for schizophrenia. Their antipsychotic medication was changed from conventional antipsychotics to one of four atypical antipsychotic drugs (olanzapine, perospirone, quetiapine, or risperidone). Patients were grouped by age (older or younger than 65 years). Subjective sleep quality and psychopathology were assessed twice: 1) at baseline, and 2) 8 weeks after switching to the atypical antipsychotic drugs. Subjective sleep quality was assessed with the Pittsburgh Sleep Quality Index (PSQI), and the Positive and Negative Syndrome Scale (PANSS) was used to measure psychopathology. RESULTS: The proportion of the patients who experienced improved subjective sleep quality was significantly higher in the elderly than in the middle-aged group. Logistic regression analysis revealed that the improvement in subjective sleep quality through administration of atypical antipsychotic drugs was predicted by increased age, daytime dysfunction, and longer sleep latency at baseline. CONCLUSION: These results demonstrate that atypical antipsychotic drugs are beneficial to the quality of sleep in elderly patients with schizophrenia. PMID- 15879587 TI - Older admissions to substance abuse treatment in 2001. AB - OBJECTIVE: The number of older people with substance abuse problems is expected to increase over the next decade. Given the expected growth in the number of elderly clients needing substance abuse treatment, the authors provide a description of admissions of patients age 55 years and over to facilities receiving some public funds. METHODS: The Treatment Episode Data Set (TEDS), a public-use data-set, contained information on 58,073 admissions to substance treatment (age 55+) and 1,043,910 admissions age 30-54 years. RESULTS: Older admissions listed only one substance-daily use of alcohol. Admission record notations indicated that these admissions were more frequently associated with income, insurance, and marriage or divorce than younger admissions. As in younger admissions, criminal justice was a major source of referral to treatment. Older patients' admissions records indicated fewer previous treatment experiences. Older male and female admissions were similar in many regards, but differed in their treatment history. The current treatment admission was more often the first for female admissions. Older female admissions were likely to be more educated than their male counterparts, with a later age at onset. CONCLUSIONS: Older admissions to substance abuse treatment differed in important ways from younger adult admissions. The older admissions tended to come from a more stable environment (income, insurance, marriage). Despite their very high frequency and amount of drinking, few of these admissions were referred to treatment by healthcare workers. PMID- 15879588 TI - Comorbid psychosocial symptoms and quality of life in patients with dementia. AB - OBJECTIVE: The authors examined the association between treatable comorbid psychosocial symptoms and quality of life (QOL) in patients with dementia. In an effort to develop a more comprehensive understanding of this relationship, this study included both patient reports of their QOL and caregiver reports of patient QOL. METHODS: Eighty-nine dyads, consisting of a community-residing veteran with dementia and his family caregiver, were interviewed once to determine the correlation between patient QOL and depression, functional disability, pain, and agitation. RESULTS: Higher patient-rated QOL was significantly associated with lower patient depression. Higher caregiver-rated patient QOL was associated with lower caregiver depression and higher patient functional status. Agitation was not related to patient-rated or caregiver-rated patient QOL. CONCLUSIONS: Authors discuss the psychometric and conceptual implications of possible differences between self- and other-ratings of QOL, and treatment implications for caregiver education interventions. PMID- 15879589 TI - The influence of depression on cognitive decline in community-dwelling elderly persons. AB - OBJECTIVE: An association between depression and cognitive decline (CD) has been observed in cross-sectional and case/control studies of elderly populations. Whereas a handful of longitudinal community studies have found depressive symptoms to predate the onset of CD, others have found no association between depression and subsequent cognitive dysfunction. The authors examined the association between depressive symptoms and cognitive errors measured 3 years apart in a longitudinal sample of community-dwelling elderly persons. METHODS: Data were obtained from 4,162 subjects assessed as part of the Established Populations for Epidemiologic Studies of the Elderly, a population survey assessing physical, psychological, and social functioning of persons 65 years and older. RESULTS: Depressive symptoms were associated with subsequent CD even after controlling for baseline cognitive status, as well as demographic (e.g., race, gender, age, and socioeconomic status) and physical functioning variables. This was true for the sample as a whole, as well as for a subsample of participants who evidenced no baseline cognitive difficulties. CONCLUSION: Study results are consistent with others finding an association between depressive symptoms and subsequent CD. Theories regarding the causal mechanisms underlying the association between depression and cognitive decline are discussed. PMID- 15879590 TI - Prevalence of depression and its correlates in Hong Kong's Chinese older adults. AB - OBJECTIVE: Because of the rapid aging of the population and inconsistent findings of previous epidemiological studies in Hong Kong, a prevalence study of depression among older adults was timely. The authors assessed the prevalence of depression among older adults and identified factors associated with it. METHODS: The authors interviewed a random representative sample of 917 community-dwelling Chinese adults age 60 and over. The 15-item Chinese Geriatric Depression Scale with a cutoff of > or = 8 was used to identify clinically significant depression in the older adults. RESULTS: The authors found that 11.0% and 14.5% of older Chinese men and women, respectively, scored above the cutoff, a prevalence rate similar to those found in other countries, including the United States, England, and Finland. Factors that were associated with an increased likelihood of depression among older adults included poor self-rated health, long-term pain, vision problems, higher level of impairment in activities of daily living, residing in Hong Kong less than 20 years, financial strain, and having less social support. CONCLUSIONS: The prevalence rate of depression among older Chinese adults in Hong Kong is more or less similar to rates found in Western countries. The data suggest that older adults who receive less social support are more likely to be depressed. PMID- 15879591 TI - Patients with very-late-onset schizophrenia-like psychosis: a follow-up study. AB - OBJECTIVE: The topic of course and outcome of very-late-onset schizophrenia-like psychosis (VLOSLP) has not received the research attention it deserves. The aim of this study was to evaluate the course of clinical symptoms and functional status of patients with VLOSLP in comparison with patients with life-long schizophrenia. METHODS: Telephone interviews were conducted on primary caregivers of 21 patients with VLOSLP who had recently been released from inpatient care. Their treating staff evaluated 21 schizophrenia inpatients according to the same criteria. RESULTS: The majority of patients with VLOSLP did not present cognitive and functional deterioration. On the other hand, 8 of the 19 patients in the elderly schizophrenia group had some functional decline; 3 of those 8 patients seemed to have some cognitive decline, as well. CONCLUSIONS: The results suggest that the VLOSLP patients present stable cognitive and everyday functioning, as compared with chronically institutionalized elderly patients with schizophrenia. PMID- 15879592 TI - Bright-light treatment reduces actigraphic-measured daytime sleep in nursing home patients with dementia: a pilot study. AB - OBJECTIVE: The authors evaluated the effects of bright-light therapy on daytime sleep among nursing home patients with dementia. METHODS: Eleven patients with sleep/wake disturbances received 2 hours/day of morning bright-light exposure for 2 weeks. Sleep was measured with actigraphy and nursing staff diaries. RESULTS: Daytime sleep decreased significantly in the period from rising time to 3:00 P.M. with bright-light treatment. Treatment effects did not last into the 16-week post treatment period. CONCLUSIONS: Bright light exposure was effective in reducing daytime sleep in nursing home patients with dementia; this finding is possibly related to bright light's acute alerting effects. PMID- 15879593 TI - Delayed onset of posttraumatic stress disorder among male combat veterans: a case series. AB - OBJECTIVE: Authors investigated the nature of delayed-onset posttraumatic stress disorder (PTSD) among combat veterans. METHODS: PTSD, along with cognitive and emotional functioning, was assessed in a case series of elderly Australian war veterans. RESULTS: Fifteen elderly male subjects consecutively referred to an outpatient psychiatric clinic were identified as having PTSD with significantly delayed onset. In most cases, the onset of PTSD symptoms was associated with unrelated medical complaints, psychosocial stress, and/or mild cognitive impairment. CONCLUSION: Environmental stressors, coupled with age-related neurodegeneration, may potentially contribute to the late-life recrudescence or emergence of PTSD symptoms in veterans exposed to combat-related trauma. PMID- 15879594 TI - Association of the serotonin transporter gene-linked polymorphic region (5 HTTLPR) genotype with depression in elderly persons after hip fracture. AB - OBJECTIVE: The authors examined the serotonin transporter gene-linked polymorphic region (5-HTTLPR) as a predictor of major depressive disorder and depressive symptoms after hip fracture, a common stressful medical event. METHODS: This was a prospective, observational study of 23 elderly rehabilitation-hospital patients during their inpatient stay. Depressive symptoms were assessed by Hamilton Rating Scale for Depression (Ham-D) and PRIME-MD. Subjects were also genotyped for 5 HTTLPR. RESULTS: Survival analysis showed that genotype significantly predicted time-to-major depressive episode. Subjects with an s allele (genotype s/l or s/s) had significantly higher Ham-D scores over 14 weeks of follow-up than those with the l/l genotype. CONCLUSION: Depressive symptoms and major depressive disorder in elderly persons after a stressful medical event may be associated with 5 HTTLPR genotype. This finding requires confirmation in a larger sample. PMID- 15879595 TI - Calcium-dependent activation of interleukin-21 gene expression in T cells. AB - Interleukin (IL)-21 is a gamma(c)-dependent cytokine produced by activated T cells with important actions for T, B, and NK cells. The IL-21 gene is adjacent to the IL-2 gene, and like IL-2, IL-21 is strongly induced at the transcriptional level after T cell activation. Interestingly, however, in contrast to the IL-2 gene, a calcium ionophore alone was sufficient to induce IL-21 gene expression in preactivated T cells. Two DNase I hypersensitivity sites were found in the IL-21 gene, corresponding to nucleotide sequences that are conserved in humans and mice. One site is located at the IL-21 promoter region and conferred T cell receptor-mediated IL-21 gene transcription. TCR-induced IL-21 gene expression was inhibited by cyclosporin A and FK506. Correspondingly, the IL-21 5'-regulatory region contains three NFAT binding sites, and induction of IL-21 promoter activity was impaired when these sites were mutated or following treatment with cyclosporin A. Thus, our studies reveal that in contrast to IL-2, a calcium signal alone is sufficient to mediate induction of the IL-21 in preactivated T lymphocytes and that this induction appears to result from specific NFAT binding. PMID- 15879596 TI - Detection of human red blood cell-bound nitric oxide. AB - Major disparities in reported levels of basal human nitric oxide metabolites have resulted in a recent literature focusing almost exclusively on methods. We chose to analyze triiodide chemiluminescence, drawn by the prospect of identifying why the most commonly employed assay in nitric oxide biology typically yielded lower metabolite values, compared with several other techniques. We found that the sensitivity of triiodide was greatly affected by the auto-capture of nitric oxide by deoxygenated cell-free heme in the reaction chamber. Potential contaminants and signal losses were also associated with standard sample purification procedures and the chemistry involved in nitrite removal. To inhibit heme nitric oxide auto-capture, we added potassium ferricyanide to the triiodide reagent, reasoning this would provide a more complete detection of any liberated nitric oxide. From human venous blood samples, we established nitric oxide levels ranging from 0.000178 to 0.00024 mol nitric oxide/mol hemoglobin. We went on to find significantly elevated nitric oxide levels in venous blood taken from diabetic patients in comparison to healthy controls (p < 0.0001). We concluded that the lack of signals reported of late by several groups using triiodide chemiluminescence for the detection of hemoglobin-bound nitric oxide may not represent levels on the border of assay sensitivity but rather underestimated values because of methodological limitations. We therefore stress the need for assay systems to be developed that differentiate between individual nitric oxide metabolite species and overcome the limitations we outline, allowing accurate conclusions to be drawn regarding physiological nitric oxide metabolite levels. PMID- 15879597 TI - Exogenously added fibroblast growth factor 2 (FGF-2) to NIH3T3 cells interacts with nuclear ribosomal S6 kinase 2 (RSK2) in a cell cycle-dependent manner. AB - Fibroblast growth factor 2 (FGF-2) has been detected in the nuclei of many tissues and cell lines. Here we demonstrate that FGF-2 added exogenously to NIH3T3 cells enters the nucleus and interacts with the nuclear active 90-kDa ribosomal S6 kinase 2 (RSK2) in a cell cycle-dependent manner. By using purified proteins, FGF-2 is shown to directly interact through two separate domains with two RSK2 domains on both sides of the hydrophobic motif, namely the NH2-terminal kinase domain (residues 360-381) by amino acid Ser-117 and the COOH-terminal kinase domain (residues 388-400) by amino acids Leu-127 and Lys-128. Moreover, this interaction leads to maintenance of the sustained activation of RSK2 in G1 phase of the cell cycle. FGF-2 mutants (FGF-2 S117A, FGF-2 L127A, and FGF-2 K128A) that fail to interact in vitro with RSK2 fail to maintain a sustained RSK2 activity in vivo. PMID- 15879598 TI - Thioredoxin reductase is irreversibly modified by curcumin: a novel molecular mechanism for its anticancer activity. AB - The thioredoxin reductase (TrxR) isoenzymes, TrxR1 in cytosol or nucleus and TrxR2 in mitochondria, are essential mammalian selenocysteine (Sec)-containing flavoenzymes with a -Gly-Cys-Sec-Gly active site. TrxRs are the only enzymes catalyzing the NADPH-dependent reduction of the active site disulfide in thioredoxins (Trxs), which play essential roles in substrate reductions, defense against oxidative stress, and redox regulation by thiol redox control. TrxRs have been found to be overexpressed by a number of human tumors. Curcumin, which is consumed daily by millions of people, is a polyphenol derived from the plant Curcuma longa. This phytochemical has well known anticancer and antiangiogenic properties. In this study we report that rat TrxR1 activity in Trx-dependent disulfide reduction was inhibited by curcumin. The IC(50) value for the enzyme was 3.6 microM after incubation at room temperature for 2 h in vitro. The inhibition occurred with enzyme only in the presence of NADPH and persisted after removal of curcumin. By using mass spectrometry and blotting analysis, we proved that this irreversible inhibition by curcumin was caused by alkylation of both residues in the catalytically active site (Cys(496)/Sec(497)) of the enzyme. However, the curcumin-modified enzyme showed a strongly induced NADPH oxidase activity to produce reactive oxygen species. Inhibition of TrxR by curcumin added to cultured HeLa cells was also observed with an IC(50) of around 15 microM. Modification of TrxR by curcumin provides a possible mechanistic explanation for its cancer preventive activity, shifting the enzyme from an antioxidant to a prooxidant. PMID- 15879599 TI - Proliferating cell nuclear antigen promotes translesion synthesis by DNA polymerase zeta. AB - DNA polymerase zeta (Pol zeta), a heterodimer of Rev3 and Rev7, is essential for DNA damage provoked mutagenesis in eukaryotes. DNA polymerases that function in a processive complex with the replication clamp proliferating cell nuclear antigen (PCNA) have been shown to possess a close match to the consensus PCNA-binding motif QxxLxxFF. This consensus motif is lacking in either subunit of Pol zeta, yet its activity is stimulated by PCNA. In particular, translesion synthesis of UV damage-containing DNA is dramatically stimulated by PCNA such that translesion synthesis rates are comparable with replication rates by Pol zeta on undamaged DNA. PCNA also stimulated translesion synthesis of a model abasic site by Pol zeta. Efficient PCNA stimulation required that PCNA was prevented from sliding off the damage-containing model oligonucleotide template-primer through the use of biotin-streptavidin bumpers or other blocks. Under those experimental conditions, facile bypass of the abasic site was also detected by DNA polymerase delta or eta (Rad30). The yeast DNA damage checkpoint clamp, consisting of Rad17, Mec3, and Ddc1, and an ortholog of human 9-1-1, has been implicated in damage induced mutagenesis. However, this checkpoint clamp did not stimulate translesion synthesis by Pol zeta or by DNA polymerase delta. PMID- 15879602 TI - Differences in baseline characteristics and in-hospital outcomes in patients with or without prior stroke undergoing percutaneous coronary intervention. AB - BACKGROUND: Percutaneous coronary intervention (PCI) is increasingly used in patients with high-risk baseline characteristics. A prior stroke may identify patients who have a higher risk for post-PCI complications. However, no comparative data exist on post-PCI outcomes of patients with or without prior stroke. METHODS: Review of a PCI database of 9,088 consecutive PCIs from July 1997 to December 2002 identified 812 PCIs in patients with a history of prior stroke and 8,044 PCIs without prior stroke. RESULTS: Patients with prior stroke had high-risk baseline characteristics [diabetes, hypertension, hyperlipidemia, smoking, peripheral arterial disease, congestive heart failure, chronic renal failure, history of prior myocardial infarction and prior coronary artery bypass graft (CABG)] and high-risk coronary anatomy (p < 0.001 for each one). The triple composite (death, myocardial infarction and emergent CABG) and the triple composite plus post-PCI stroke were higher in patients with prior stroke (11.2% vs. 4.8%; p < 0.001; z = 7.617 and 12.1% vs. 5.0%; p < 0.001; z = 8.271, respectively. CONCLUSION: Patients with prior stroke constitute a high-risk PCI cohort with higher rates of in-hospital adverse events. A prior stroke history should be considered in evaluating potential candidates for PCI. PMID- 15879603 TI - New heparin dosing regimen for diabetics undergoing percutaneous coronary intervention. AB - It has been previously demonstrated that diabetics are less sensitive to heparin compared to non-diabetics. We hypothesized that an initial heparin dose of 80 IU per kilogram administered to diabetics rather than 70 IU per kilogram might yield a more optimal initial ACT of 300 to 350 seconds when glycoprotein IIb/IIIa receptor antagonists are not used. We prospectively studied 130 elective PCI patients without diabetes treated with 70 IU per kilogram of unfractionated heparin and 81 elective PCI patients with diabetes treated with 80 IU per kilogram, and compared the initially achieved ACT. The mean heparin dose given per kg was greater (by intention) in diabetics versus non-diabetics. Despite that, there was no significant difference in the initially achieved ACT in diabetics and non-diabetics. PMID- 15879604 TI - Long-term outcomes based on time-to-angioplasty in patients admitted with non-ST segment elevation acute coronary syndromes. AB - OBJECTIVE: We investigated the impact of the duration from hospital admission to coronary angiography on the outcome of patients admitted with non ST-segment elevation acute coronary syndromes (NSTE-ACS). BACKGROUND: Invasive risk stratification in patients with acute coronary syndromes (ACS) has been shown to improve outcome in contemporary studies. It is unclear whether early coronary angiography is better than initial medical therapy with later angiography. METHODS: We performed an analysis of patients admitted to a tertiary coronary intensive care unit (CICU) with NSTE-ACS and had coronary angiography performed during the same hospitalization. Patients were categorized into three groups based on the time-to-angiography: same-day, 1 to 2 days, and > 2 days. The baseline clinical features, angiography results, 30-day, 6-month cardiovascular outcome and 3-year mortality rate were compared between the groups before and after adjusting for confounding variables. RESULTS: A total of 836 fulfilled the inclusion criteria. Patients undergoing angiography > 2 days had a higher incidence of 3-vessel disease (45.7% vs. 31.7%, p < 0.001), underwent less percutaneous interventions at the time of the angiography (41.6% vs. 56.7%, p < 0.001), and more frequent coronary artery bypass surgery (9.9% vs. 15.3%, p = 0.05). Patients undergoing late invasive risk stratification (> 2 days) had increased 3-year mortality (OR 2.12, 95% CI 1.03-4.35, p = 0.04) after adjusting for confounding variables. CONCLUSION: In patients with NSTE-ACS and no contraindication to angiography, delayed angiography of more than 2 days of presentation was associated with increased mortality at 3 years. PMID- 15879605 TI - Differential outcome after intracoronary radiation therapy is related to a simple classification based on lesion length and reference diameter. AB - OBJECTIVE: We sought to develop a prognostic lesion classification based on simple angiographic parameters, lesion length and reference diameter that predicts differential outcome in patients undergoing intracoronary radiation. METHODS AND RESULTS: Three types of lesions were identified: Type A characterized by lesion length less than or equal to 30 mm, reference diameter > 2.5 mm to less than or equal to 4.0 mm (short lesion: normal diameter), Type B by lesion length less than or equal to 30 mm, reference diameter less than or equal to 2.5 mm or > 4 mm (short lesion: extreme diameter), and Type C by lesion length > 30 mm (long lesion). A total of 1,151 lesions (77.7% in-stent restenosis) in 1,098 consecutive patients undergoing brachytherapy were classified into these 3 lesion types. Overall, 79.9%, 10.3% and 9.8% patients met the criteria for Type A, B and C lesions. While the in-hospital major adverse cardiac event (MACE) rate was 1.4%, 3.6% and 3.8% (p = 0.026), the 6-month MACE rate was 16.1%, 22.5% and 32.1% (p < 0.001), the angiographic restenosis rate was 21.3%, 32.4% and 42.4% (p < 0.001), and the late thrombosis rate was 4.1%, 9.0% and 11.3%, (p < 0.001) in Type A, B and C lesions, respectively. Consequently, with increasing lesion severity, 3 risk groups with low, medium and high risk were defined. Multivariate analysis showed that Type B and C lesions were independent predictors of 6-month MACE (OR, 1.5 and 1.9, respectively). CONCLUSION: The proposed novel and easily applicable lesion classification effectively predicts early and medium term outcome, and may be used for appropriate therapeutic decision making in patients undergoing brachytherapy. PMID- 15879606 TI - Catheter closure of perforated secundum atrial septal defect under intracardiac echocardiographic guidance using a single amplatzer device: feasibility of a new method. AB - OBJECTIVES: Little is known about the feasibility and effectiveness of perforated secundum atrial septal defects (ASDs) transcatheter closure. This study tested whether this type of fenestration can be percutaneously closed using a single Amplatzer PFO or Cribriform Occluder device in a patch-like fashion under intracardiac echocardiographic (ICE) guidance and monitoring. METHODS: Twenty four adult patients with perforated ASDs underwent transcatheter single Amplatzer PFO or Cribriform Occluder device closure by using ICE as the primary imaging tool, both for crossing the flap valve and monitoring each stage of the procedure. The entire atrial septum and fossa ovalis end diastolic dimensions on 2 orthogonal ICE planes, as well as the distance between the eccentric guidewire passage and the more adjacent rim of the fossa ovalis on the longitudinal ICE plane, have been measured. RESULTS: On the basis of the ICE fossa ovalis size and its geometric evaluation, in order to completely cover the fossa ovalis with a single device, 2 Amplatzer PFO Occluders 25 mm, 9 Amplatzer PFO Occluders 35 mm, and 13 Amplatzer Cribriform Occluders (four 25 mm and nine 35 mm) have been implanted successfully. During follow-up (31.4, 7.2 months), complete closure by contrast echo color Doppler occurred in 16/24 (67%) patients after 24 hours, 19/24 (79%) after 1 mouth, 20/24 (83%) after 3 months, 22/24 (92%) after 1 year, and 23/24 (96%) after 2 years. CONCLUSIONS: Transcatheter closure of perforated ASDs using a single Amplatzer Occluder device under ICE guidance appears to be a suitable procedure, providing a low rate of residual shunting. PMID- 15879607 TI - Eosinophilic arteritis with coronary aneurysms and stenoses. PMID- 15879608 TI - Use of a sirolimus-eluting stent to treat failure of a nonpolymer release paclitaxel-eluting stent implanted for in-stent restenosis. PMID- 15879609 TI - First experience with intra-renal fenoldopam in a patient with heart failure. PMID- 15879610 TI - Successful use of a new guidewire with radiofrequency ablation capability for the treatment of chronic total occlusion at the ostium of the left anterior descending artery. PMID- 15879611 TI - A case of late stent thrombosis after exercise electrocardiography: exercise- or residual plaque-induced? PMID- 15879612 TI - Issues in the management of antiplatelet therapy in patients undergoing surgical revascularization. A roundtable discussion. PMID- 15879613 TI - Acute myocardial infarction: early strategies to optimize results. PMID- 15879614 TI - The enigma of primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a chronic cholestatic disease characterized by a striking predominance in female patients (with most cases diagnosed between ages 40 and 60 yr) as well as serum auto-antibodies to mitochondrial antigens, elevated serum immunoglobulin M,progressive destruction of intrahepatic bile ducts, and, ultimately, liver cirrhosis and failure(1). The precise mechanisms leading to selective destruction of biliary epithelial cells lining intrahepatic bile ducts are still unknown, although numerous immunomediated pathways have been proposed. Genetic background appears to be important in determining susceptibility to the disease (2), but no clear association with alleles in the major histocompatibility complex has been identified. Molecular mimicry either by infections (3) or xenobiotics (4) has been proposed to be capable of breaking tolerance in genetically predisposed individuals, thus leading to onset of PBC. This article describes and discusses the available data regarding the immunomediated pathogenesis of PBC (with particular attention to auto-antibodies and autoreactive T-cells) and presents the recent evidence indicating a role for either xenobiotic chemicals or novel infectious agents in the induction of the disease. PMID- 15879615 TI - Mitochondrial antigens as targets of cellular and humoral auto-immunity in primary biliary cirrhosis. AB - Several factors point toward an auto-immune pathogenesis for primary biliary cirrhosis(PBC), mostly based on the presence of serum auto-antibodies to mitochondrial antigens(AMAs) and autoreactive T cells (both helper and cytotoxic). Interestingly, epitopes recognized by AMA and T-cell clones are located within overlapping areas of the antigens. Moreover,a role for an imbalance in cytokine pattern and for natural-killer lymphocytes has also been proposed. Despite several experimental reports, no clear evidence is available regarding the interaction of these factors leading to bile duct destruction. This article reviews the current reports regarding the auto-immune reaction against mitochondrial auto-antigens in PBC. PMID- 15879616 TI - Immunopathogenesis of primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease of unknown etiology;however, lymphocytic portal tract infiltration is suggestive of an immune-mediated basis for PSC. Associations with inflammatory bowel disease- especially ulcerative colitis--and with other auto-immune diseases, together with genetic associations, further suggest that PSC may be an immune-mediated disease. The immunogenetics of PSC have been the subject of active research, and several human leukocyte antigen (HLA)- and non-HLA-associated genes have been implicated in the development of the disease. Lymphocytes derived from the inflamed gut may enter the liver via the enterohepatic circulation to cause hepatic disease.PSC may be triggered in genetically susceptible individuals by infections or toxins entering the portal circulation through a permeable colon and, therefore, evoking an abnormal immune response. PMID- 15879617 TI - Overlap syndromes of cholestatic liver diseases and auto-immune hepatitis. AB - Approximately 18% of patients with auto-immune liver disease present with features characteristic of a second auto-immune liver disease. These cases have been termed "overlap syndromes."The pathogenesis of overlap syndromes is poorly understood, and few data are available regarding the clinical characteristics and outcome of this disease. Therefore, a consensus on the definition of overlap syndromes has not been reached. A common genetic background between auto-immune hepatitis (AIH) and its overlap with primary biliary cirrhosis(PBC) or primary sclerosing cholangitis (PSC) might confer susceptibility to a more inflammatory phenotype, probably requiring combined immunosuppressive treatment. This article focuses on the prevalence, diagnosis, and treatment of the overlap syndrome of AIH and PBC or PSC. PMID- 15879618 TI - Role of auto-antibodies for the diagnosis of chronic cholestatic liver diseases. AB - Auto-antibodies are an integral part of the diagnostic armentarium in chronic cholestatic liver disorders, such as primary sclerosing cholangitis (PSC), primary biliary cirrhosis (PBC),auto-immune cholangitis, or overlap syndromes among these disorders. However, care should be taken not to overestimate the diagnostic specificity. Auto-antibodies to mitochondrial antigens(AMAs) with reactivity to the E2 subunit of the pyruvate dehydrogenase complex represent the hallmark antibody for the diagnosis of PBC, whereas antinuclear antibodies (ANAs)with low disease specificity are found in up to 50% of these sera. Antibodies that recognize nuclear envelope proteins exert a similarly high diagnostic specificity as AMA in PBC but occur at a rather low prevalence. The role of auto-antibodies is less well-studied for patients with PSC, but there is growing evidence that only antineutrophil cytoplasmic antibodies(ANCAs) are of relevant diagnostic significance. In contrast, auto-antibodies-particularlyAMAs do not contribute to the diagnosis of auto-immune cholangitis, whereas ANCAs,ANAs, smooth muscle antibodies, and AMAs are of varying significance in PBC auto-immune hepatitis (AIH) or PSC-AIH overlap syndromes. It has been widely accepted that the course of the auto-antibody serum end point titers are not suited for the clinical management of patients with chronic cholestatic liver disorders. Additionally, auto-antibodies in these disorders usually do not contribute to the immunopathogenesis of the disease. PMID- 15879619 TI - Medical treatment of primary biliary cirrhosis and primary sclerosing cholangitis. AB - Chronic cholestasis is the main feature of primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC), the most common chronic cholestatic liver diseases in adults. Although the etiology of both diseases remains poorly understood, auto-immune processes appear to be important, particularly in PBC. PBC and PSC usually slowly progress to cirrhosis,liver failure, and death, unless liver transplantation is performed. Ursodeoxycholic acid(UDCA), a hydrophilic dihydroxy bile acid, is the only drug currently approved for the treatment of patients with PBC and is also used in patients with PSC. In addition to UDCA, patients with PSC should be referred to endoscopic dilatation of major bile duct stenoses. Several potential mechanisms of action of UDCA have been proposed, including intracellular modulation of signaling events and secretion. Various immunosuppressive drugs have been evaluated alone or in combination with UDCA especially for the treatment of PBC. Of these drugs,the topical corticosteroid budesonide, together with UDCA, appears promising in the treatment of early stage PBC, but data remain insufficient to warrant use of budesonide outside of controlled studies. PMID- 15879621 TI - Endoscopic treatment of dominant stenoses in patients with primary sclerosing cholangitis. AB - Primary sclerosing cholangitis is characterized by progressive fibrosing inflammation of the bile ducts, leading to their obliteration, which results in cholestasis and, finally, cirrhosis of the liver. Over time, the majority of patients with advanced disease develop dominant stenoses of major bile ducts. Ursodeoxycholic acid (UDCA) treatment does not prevent the development of such stenoses. Endoscopic measures allow the opening of short- and long-segment stenoses of the common bile duct and also of short segment stenoses of the hepatic ducts. Inpatients treated by early endoscopic dilatation of dominant stenoses, as well as with UDCA,survival may be significantly improved (compared with the predicted survival). PMID- 15879620 TI - Extrahepatic manifestations of cholestatic liver diseases: pathogenesis and therapy. AB - Pruritus, fatigue, and metabolic bone disease are frequent complications of cholestatic liver diseases, which can be quite distressing for the patient and can considerably reduce the quality of life. The molecular pathogenesis of these extrahepatic manifestations of cholestasis is poorly understood, and hypotheses to explain these symptoms are being discussed. This article provides treatment recommendations for the complications of cholestasis based on putative pathomechanisms and summarizes recent experimental and clinical data involving management options. PMID- 15879622 TI - Surgical treatment of primary biliary cirrhosis and primary sclerosing cholangitis. AB - Primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) are progressive cholestatic liver diseases of supposed auto-immune etiology. The clinical course is unpredictable and, in many patients, leads to end-stage liver disease or a poor quality of life. Conservative therapy only has a limited effect on the natural history, but orthotopic liver transplantation(OLT) offers a definitive therapeutic option. Retrospective analysis was performed for 38 patients with PBC and 17 patients with PSC who underwent OLT between January 1986 and June 2003 at our institution. Median followup after OLT was 72 mo.Cumulative survival at 5 yr post-OLT was 84% in the PBC group and 73% in the PSC group. Compared with OLT for other benign diseases, actuarial survival rates at 5 and 10 yr post-OLT were significantly better for patients with PBC, whereas there was no difference in survival after OLT for patients with PSC. Survival rate at 5 yr post-OLT was significantly increased for patients with PBC who had a Child-Pugh B liver cirrhosis (93%) compared with those who had Child-Pugh C cirrhosis (60%). Retransplantation rate was 18.2% (resulting from biliary complications in three cases). Surgical techniques had no effect on outcome after OLT in both groups. We concluded that liver transplantation represents a safe and beneficial therapy for patients with end-stage PBC. Cirrhotic patients with PSC also benefit from OLT, with an outcome comparable to that of liver cirrhosis of other etiologies. PMID- 15879623 TI - Antitumor effect of a combination of lysine, proline, arginine, ascorbic acid, and green tea extract on pancreatic cancer cell line MIA PaCa-2. AB - BACKGROUND: Current treatment of pancreatic cancer is generally associated with poor prognosis, even if diagnosed early, owing to its aggressive rate of metastasis and non-responsiveness to chemotherapy and radiotherapy. Matrix metalloproteinases (MMPs) have received much attention in recent years for their role in various malignancies, and have been implicated in tumor invasion, metastasis, and angiogenesis. AIM OF STUDY: Reported antitumor properties of ascorbic acid, lysine, proline, and green tea extract prompted us to investigate the effect of a combination of lysine, proline, arginine, ascorbic acid, and green tea extract on pancreatic cancer cell line MIA PaCa-2 for viability, MMP expression, invasion, and morphology. METHODS: Viability was evaluated based on cell proliferation by MTT assay and MMP expression in condition media by gelatinase zymography. Invasion through Matrigel was assayed and morphology was observed by hematoxylin and eosin (H+E)staining. Data was analyzed by independent sample "t" test. RESULTS: The nutrient mixture (NM) did not inhibit cell proliferation at 10 microg/mL and exhibited a dose-dependent antiproliferative effect with maximum inhibition of 38% over the control at 1000 microg/mL. Zymography demonstrated production of only MMP-9, which showed a dose-dependent decreased expression that was abolished at 100 microg/mL of NM. Invasion through Matrigel was inhibited at 10, 50, 100, and 500 microg/mL by 66%, 66%, 87% and 100%, respectively. H&E staining did not indicate changes even at the highest concentration of NM. CONCLUSION: Our results suggest that the formulation of green tea extract, lysine, proline, and ascorbic acid, tested as a promising adjunct to standard treatment of pancreatic cancer, by inhibiting MMP expression and invasion without toxic effects important parameters in cancer metastasis. PMID- 15879624 TI - A phase II study of capecitabine plus oxaliplatin (XELOX): a new first-line option in metastatic colorectal cancer. AB - BACKGROUND: Capecitabine and oxaliplatin are both effective and well-tolerated monotherapies for the treatment of advanced colorectal cancer (CRC). Oxaliplatin has also been shown to be very effective when combined with 5-FU/LV in the first line setting. AIM OF THE STUDY: Assess the efficacy and safety of capecitabine plus oxaliplatin (XELOX) in patients with previously untreated advanced CRC. METHODS: Fifty-three patients with measurable disease received capecitabine 1,000 mg/m2 twice daily on d 1-14 and oxaliplatin 130 mg/m2 on d 1, every 3 wk. Of these, 52 were evaluable for safety and 49 for antitumor response. RESULTS: There was a low rate of grade 1/2 adverse events; grade 3/4 events included leukopenia (10%), neutropenia (6%), thrombocytopenia (2%), nausea/vomiting (4%), and diarrhea (4%). The overall response rate was 39% (95% CI, 25-54%) and median time to disease progression was 7.8 mo. CONCLUSIONS: XELOX is an active and well tolerated first-line treatment for advanced CRC. Randomized phase III studies are ongoing to compare XELOX with FOLFOX in view of the comparable efficacy and safety but superior convenience of XELOX therapy. PMID- 15879625 TI - Biological similarities and differences between pancreatic intraepithelial neoplasias and intraductal papillary mucinous neoplasms. AB - BACKGROUND: Ever since the classification of pancreatic intraepithelial neoplasia (PanIN) was published, studies on the precursor lesions of pancreatic cancer have been advancing along a new directions, using standardized terminology. There are few studies that have examined the biological differences between PanIN and intraductal papillary mucinous neoplasm (IPMN) in detail. AIMS: PanIN and IPMN, which are similar in morphology, were compared using various indicators, with the aim of identifying the similarities and differences between the two. METHODOLOGY: A total of 46 PanINs and 37 ducts with IPMN were identified in 19 patients with invasive ductal carcinoma and 18 patients with IPMN. These PanINs and IPMNs were examined immunohistologically with respect to the expression patterns of HER2/neu, DPC4/Smad4, Akt/PKB, p53, cyclin A, Ki67, MUC1, and MUC2. RESULTS: Significant differences in the expression of MUC1 and MUC2 were observed between IPMNadenoma and PanIN-2 and between CIS and PanIN-3 (MUC1: p = 0.001 and p = 0.005, respectively; MUC2: p = 0.002 and p < 0.001, respectively). A significant difference in the p53 expression level was also observed between CIS and PanIN-3 (p = 0.015). CONCLUSIONS: In both IPMN and PanIN, the grade of atypism increased with increasing expression of HER2/neu, DPC4/Smad4, and Akt/PKB, along with progression in the process of multistage carcinogenesis. Although the expression levels of these factors reflected the grade of atypism, they did not reflect any differences in the grade of biological malignancy between IPMN and PanIN. On the other hand, MUC1 and MUC2 may serve as indicators of the direction of differentiation, i.e., either progression to IDAC or IPMN. Positivity for MUC1 was believed to suggest differentiation into IDAC, and positivity for MUC2 appeared to be indicative of differentiation into IPMN. Such indication of the direction of differentiation seemed to appear in PanIN1-2, even before abnormalities of HER2/neu, Akt/PKB, DPC4/Smad4, p53, and cyclin A expression began to be detected. PMID- 15879626 TI - Impact of a family history of colorectal cancer on age at diagnosis, anatomic location, and clinical characteristics of colorectal cancer. AB - BACKGROUND: Among the risk factors for colorectal cancer (CRC) is a family history of colorectal cancer. Reliable evidence is needed regarding the clinical characteristics of cancer in patients with this history to determine if a change in the diagnostic approach is needed. AIM OF THE STUDY: This study set out to determine specific clinical outcomes in patients with CRC with a family history of one first-degree relative with sporadic colorectal cancer compared to control patients with colorectal cancer but without the family history. METHODS: We designed a case-control study of colorectal cancer registry data between 1988 and 1999. Patients with a family history of one first-degree relative with colorectal cancer were compared to those without the history with regard to four characteristics: age at cancer diagnosis, anatomic location of the cancer, presence of distal adenomas with proximal cancer, and stage of disease at diagnosis. RESULTS: Nine hundred and twenty-one patients met the inclusion criteria. Family history was positive in 124 patients. The demography of the populations was similar, except for mean age, which was 65 yr for men with a family history and proximal cancer compared to 70 yr for their counterparts without the family history (p = 0.03). The anatomic location of the cancer, presence of distal benign neoplasia when the cancer was proximal, and disease stage at diagnosis were not different between the groups. CONCLUSIONS: Men with a family history of sporadic colorectal cancer and proximal colon cancer were younger than men without the family history and proximal colon cancer. The overall results do not indicate that a change in the diagnostic approach is needed. PMID- 15879627 TI - Bowel obstruction in patients with metastatic cancer: does intervention influence outcome? AB - BACKGROUND: Both surgical and nonsurgical options are available to treat bowel obstruction in patients with metastatic cancer. The goal is straightforward: to restore bowel patency and palliate the symptoms of obstruction. Yet the most appropriate management is often a challenging decision. AIM OF THE STUDY: We sought to review our experience in managing patients with metastatic cancer and bowel obstruction. METHODS: A retrospective review was performed to identify all patients admitted at University of Wisconsin Hospital between 1993 and 2000 with the diagnoses of both bowel obstruction and metastatic cancer. Demographic data, type of management, postoperative complications, and outcome were analyzed. RESULTS: A total of 114 patients with primarily colorectal or gynecologic malignancies were identified. Patients' first bowel obstructions were managed in one of two ways: (1) definitive surgical intervention (n = 47), or (2) conservative management (n = 67). The median overall survival was 3 mo for the entire study group. There was no significant difference in overall or obstruction free survival based on management, presence of recurrent bowel obstruction, or type of primary cancer. The only factor that was significant in predicting poor overall survival included a disease-free interval of less than 1 yr (time of diagnosis of primary cancer to time of bowel obstruction, p = 0.002). CONCLUSIONS: Bowel obstruction in patients with metastatic cancer is a terminal event, with a 3-mo median survival. Because there is no difference in overall or obstruction-free survival based on management, the treatment for palliation of bowel obstruction in patients with metastatic cancer should be individualized. PMID- 15879628 TI - HPV-associated carcinoma of esophagus in the young: a case report and review of literature. AB - Esophageal carcinoma (EC) is rare in the younger age group, with only 15 cases reported in world literature. It is even rarer for an etiological agent to be implicated.A case of a 15-yr-old boy with squamous cell carcinoma of the esophagus is reported. Human papillomavirus (HPV) 16 DNA sequences were detected in the neoplasm and the adjacent mucosa by polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP). The adjacent mucosa also showed virus-induced changes. PMID- 15879629 TI - Primary malignant fibrous histiocytoma of the jejunum metastatic to the spine: report of a case. AB - We report a case of malignant fibrous histiocytoma (MFH) metastatic to the spine. A 41-yr-old male was admitted to our hospital for radiation treatment of MFH of the spine. He began to show signs suggestive of partial small bowel obstruction. Computed tomography demonstrated jejuno-jejunal intussusception. The patient was taken to the operating room, where the diagnosis was confirmed. Partial jejunal resection was performed. The lead point of the intussusception was histologically diagnosed to be a high-grade malignant fibrous histiocytoma. We believe that the spinal lesion was the metastatic lesion and that metastasis occurred via the vessels of Adamciewicz. To our knowledge this is the first case thus reported. PMID- 15879630 TI - Recurrence of primary hepatic carcinoid tumor in the remnant liver 13 yr after resection. AB - We report here a case of primary hepatic carcinoid tumor (PHCT) recurring in the remnant liver 13 yr and 10 mo after first resection. A 70-yr-old man developed four hypervascular tumors in the liver in December 2003. He had undergone curative left-lobe hepatectomy for PHCT in February 1990. Histopathological examination of the tumor biopsy specimen showed that the tumor was composed of uniform round-to-oval cells with solid arrangement and the tumor cells stained positive for chromogranin A, synaptophysin, and neuron-specific enolase. We diagnosed this case as an intrahepatic metastasis of PHCT with a long latency period, based on the fact that no primary site of carcinoid tumor could be found despite intensive examination and the immunohistochemical findings of the resected tumors were essentially same as those of PHCT in 1990. Although PHCT is reported to have a more favorable prognosis than other hepatic cancer or metastatic carcinoid tumor in the liver, long-term observation is recommended. PMID- 15879631 TI - Gallbladder carcinoma presenting as exfoliative dermatitis (erythroderma). AB - Although exfoliative dermatitis (erythroderma) secondary to malignancy is commonly associated with lymphomas or leukemias, coincident gastrointestinal (GI) malignancy and erythroderma is rare. The authors recently encountered a patient with gallbladder carcinoma presenting as erythroderma. A 77-yr-old Japanese man presented with a 3-mo history of erythematous eruptions with pruritus over almost the entire body. After confirming the diagnosis of erythroderma, asymptomatic gallbladder carcinoma was found. Further investigations detected no malignancies in other organs. An extended cholecystectomy was performed. Histologic examination of resected specimens revealed poorly differentiated adenocarcinoma with negative resection margins. The eruptions with pruritus resolved within 1 wk after the operation. This is the first report, to our knowledge, of coincident biliary malignancy and erythroderma. The experience of the current patient suggests that erythroderma secondary to GI malignancy may resolve spontaneously after curative resection of the tumor. PMID- 15879632 TI - Zollinger-Ellison phenotype in the absence of hypergastrinemia and islet-cell tumor. AB - Patients with the Zollinger-Ellison syndrome are characterized by islet-cell tumors, striking gastric acid hypersecretion, and peptic ulcer disease. They often experience severe abdominal pain, diarrhea, and gastrointestinal bleeding with potentially life-threatening consequences. It is a rare syndrome caused by non-beta cell islet-cell tumors (gastrinomas) located in or in proximity to the pancreas. These tumors freely secrete gastrin, a peptide hormone that serves as a powerful stimulant of gastric acid secretion. Exuberant secretion of gastrin from the gastrinomas produces severe gastric acid hypersecretion that often leads to impressive peptic ulcer disease and the constellation of symptoms listed above. We describe a patient presenting with clinical manifestations characteristic of the ZES with strikingly elevated gastric acid secretion,multiple ulcers in the first and second portions of the duodenum and diarrhea, but in absence of islet cell tumor and/or hypergastrinemia. PMID- 15879633 TI - Cut-off levels based on deviation from standard negative control is better than moderate level based on fixed cut-off for ACA assessed using ELISA for the diagnosis of antiphospholipid syndrome. AB - AIM: To assess the consistency of the standard negative control of IgG and IgM ACA levels within runs and batches of tests, and levels of ACA agreement between those established according to deviation from standard negative control and those established based on a fixed level cut off. METHODS: Serum samples of 148 patients who presented an INR < 0.9 or prothrombin activity of > 130% or aPTT below 0.8 times control or thrombosis with aPTT below 1.2 times control were tested in a 22-time running test to determine IgG and IgM ACA levels using Quanta Lite ACA IgG (HRP) and Quanta Lite ACA IgM (HRP) commercial reagents. RESULTS: Coefficients of variant within runs and batches of standard negative control IgG and IgM ACA levels were 19.30% and 29.17% respectively. Using kappa statistics to determine degree of agreement between cut-off levels by deviation from standard negative control and fixed cut-off level of ACA identified using ELISA, the disagreement in IgM and IgG were k 0.30, and 95% CI of k 0.27 to 0.34 (z = 1.033, p = 0.3015), and k 0.63, and 95% CI of k 0.53 to 0.73 (z = 1.411, p = 0.1584) for cut-off levels based on deviations from standard negative control and fixed cut off levels respectively. Cut-off levels based on deviation from standard negative control was more sensitive, with a 92% predictive true positive value, compared to a 69% predictive true positive value by fixed cut-off levels of IgM ACA detected using ELISA, and nearly equivalent to IgG ACA, with 84.4% and 87.1% predictive true positive values respectively. CONCLUSION: Cut-off points based on fixed levels of ACA detected using ELISA cannot be applied, because both IgG and IgM ACA levels of standard negative control were inconsistent among runs and batches. Cut-off points based on the deviation of 3 standard negative control levels for IgG ACA and based on deviations of 2.5 times from standard negative control levels for IgM ACA were better than cut off by fixed levels of ACA in producing true positive results. PMID- 15879634 TI - The diagnostic value of combined risk factor analysis and radiological imaging in determining osteoporosis in post-menopausal women. AB - AIM: To determine the diagnostic value of risk factor analysis (age, duration of menopause, body mass index and physical activities) and radiological imaging (Singh index and cortical index of the femoral neck) in diagnosing osteoporosis in post-menopausal women. METHODS: The study was cross sectional on 64 post menopausal women without secondary risk factor for osteoporosis. They were classified proportionally using the Singh index. Bone density was measured using DEXA (dual x-ray absorptiometry) on the femoral neck and lumbal 2-4 spine areas. The Singh index and cortical index of the femoral neck were evaluated using femoral neck antero-posterior x-ray. Physical activities were measured using a Historical leisure activity questionnaire. Bivariat statistical analysis was conducted using the t-test and chi-square, whereas multivariate analysis was conducted using multinomial logistic regression. RESULTS: There was a significant association (p<0.05) between bone density and age, body weight, height, body mass index, duration of menopause and Singh index. With multinomial logistic regression analysis, it was demonstrated that only Singh index, the duration of menopause and body mass index had the highest sensitivity and specificity. The score system algorithm could be utilized in two steps, the first was to diagnose osteoporosis and the second was to distinguish between osteopenia and normal bone. This score system had a sensitivity of 91.4% and a specificity of 89.6%, a positive prediction value of 91.4% in determining osteoporosis, and a sensitivity of 66.7%, a specificity of 89.1% and a positive prediction value of 70.6% in determining osteopenia, whereas the negative prediction value was 75%. CONCLUSION: The score system algorithm is the best method for determining osteoporosis in post-menopausal women. If there is osteopenia, evaluation using DEXA is then required. The score system algorithm cannot be used to follow up the therapy. PMID- 15879635 TI - Hepatic cirrhosis caused by Caroli disease. PMID- 15879636 TI - Diagnostic problems of pleuroperitoneal tuberculosis. PMID- 15879637 TI - Heat stroke. PMID- 15879638 TI - The role of aromatase inhibitor (AI) in the treatment of breast cancer. PMID- 15879639 TI - Toxoplasma encephalitis in HIV-infected person. PMID- 15879640 TI - Epidemiology of asthma in Indonesia. PMID- 15879641 TI - Allergy skin testing in clinical practice. PMID- 15879642 TI - An outbreak of Campylobacter jejuni enteritis in a school of Madrid, Spain. AB - An outbreak of gastroenteritis caused by Campylobacter infection was identified in May 2003 in a school in Madrid, Spain. Eighty one cases were identified in a total of 253 people studied. A retrospective cohort study showed that a custard made with ultra high temperature (UHT) milk was associated with illness (RR: 3.15; 95% CI: 1.25-7.93). The custard was probably contaminated with Campylobacter jejuni from a raw chicken prepared a day previously in the same kitchen. Our recommendations were to periodically remind the school s authorities how to act if an outbreak should be suspected, to include the monitoring of a food handler s working day in each environmental investigation in order to detect any risk behaviour; to implement microbiological analysis from the surfaces and utensils of the collective kitchens and improve the sanitary education of food handlers. PMID- 15879643 TI - Infectious diseases surveillance activities in the north of Portugal, during the EURO 2004 football tournament. AB - A European football tournament (EURO 2004) took place in Portugal, from the 12 June to the 4 July 2004. Portugal's Northern Regional Health Authority serves a population of 3.2 million people. This region hosted 12 matches, more than any other region. We describe the communicable disease surveillance activities in the region, during EURO 2004. Ten foodborne outbreaks, seven cases of meningococcal disease and one case of legionnaires disease, were detected. Visitors were not affected, furthermore, cases among residents seemed not to be influenced by the presence of thousands of visitors. A similar pattern has been observed at other mass gatherings where special surveillance activities were implemented. This does not reduce the importance of public health surveillance during such mass gatherings. Furthermore, evaluation of this special activities should be an opportunity to put, issues of communicable disease surveillance resources, priorities, organisation and training back on the agenda. PMID- 15879644 TI - Surveillance of Lyme borreliosis in Germany, 2002 and 2003. AB - Lyme borreliosis is a potentially serious infection common in Germany, but little data about its incidence, distribution, and clinical manifestations are available. Lyme borreliosis is not a notifiable disease in Germany, but six of Germany's 16 states - Berlin, Brandenburg, Mecklenburg-Vorpommern, Sachsen, Sachsen-Anhalt and Thuringen, have enhanced notification systems, which do include Lyme borreliosis. The efforts made in these states to monitor confirmed cases through notification are therefore an important contribution to understanding the epidemiology of Lyme borreliosis in Germany. This report summarises the analysis of Lyme borreliosis cases submitted to the Robert Koch Institut during 2002-2003. The average incidence of Lyme borreliosis of the six East German states was 17.8 cases per 100,000 population in 2002 and increased by 31% to 23.3 cases in 2003, respectively. Patient ages were bimodally distributed, with incidence peaks among children aged 5- 9 and elderly patients, aged 60- 64 in 2002, and 65- 69 in 2003. For both years, 55% of patients were female. Around 86% of notified cases occurred from May to October. Erythema migrans affected 2697 patients (89.3%) in 2002 and 3442 (86.7%) in 2003. For a vector-borne disease, like Lyme borreliosis, the risk of infection depends on the degree and duration of contact between humans and ticks harbouring Borrelia burgdorferi. As infectious ticks probably occur throughout Germany, it is likely that the situation in the remaining 10 German states is similar to that of the states in this study. PMID- 15879645 TI - A medical locum service as a site for sentinel influenza surveillance. AB - Surveillance of influenza in the countries of the European Union includes a sentinel network of general practitioners reporting cases of influenza-like illness (ILI), the collection of specimens for virological testing, and laboratory reporting of influenza diagnoses. In Victoria, Australia we have a similar sentinel surveillance system, with ILI defined by fever, cough and fatigue, and influenza seasons described by thresholds. The coordination of sentinel general practices can, however, be time consuming and expensive. For the last two influenza seasons we have used a deputising medical locum service as a sentinel site for influenza surveillance. We are not aware of such a service being used as a sentinel site elsewhere in the world. In both retrospective and prospective comparisons, we have shown that ILI surveillance patterns from sentinel general practices are very similar to those from the locum service. Because of its timeliness, flexibility, patient mix and geographic spread, locum service surveillance is able to supplement sentinel ILI surveillance and may also have a role in the recognition of emerging disease patterns. This is likely to be true not only in Australia but also in countries of the European Union. PMID- 15879646 TI - Epidemiological and virological assessment of influenza activity in Europe during the 2003-2004 season. AB - The 2003-2004 influenza season in Europe was dominated by the spread of the new drift variant A/Fujian/411/2002 (H3N2)-like virus which was not perfectly matched with the A(H3N2) component of the influenza vaccine. Sporadic cases of this virus were detected in Europe at the end of the 2002-2003 season and influenza activity associated with this virus began relatively early during the 2003-2004 season. Generally, influenza activity first occurred in the west of Europe (Ireland, the United Kingdom and the Iberian Peninsula) in October/November and gradually moved east across Europe, affecting Latvia, Lithuania and Poland during the months of January and February 2004. In general, the intensity of clinical activity was higher than during the 2002-2003 season (in 13 out of 20 networks) and, in countries reporting age specific data, the highest consultation incidences were observed among children aged 0-14. However, despite the emergence of the A(H3N2) drift variant, clinical incidences were not especially high compared with historical data. The composition of the 2004-2005 influenza vaccine has been modified compared with the 2003-2004 season and includes an A/Fujian/411/2002 (H3N2)-like virus strain and a new B virus strain (a B/Shanghai/361/2002-like virus). PMID- 15879647 TI - Cholestatic jaundice during infancy. PMID- 15879648 TI - Relationship between serum alanine aminotransferase levels and liver histology in chronic hepatitis C-infected patients. AB - BACKGROUND: The efficacy of serum alanine aminotransferase (ALT) levels in predicting the severity of hepatitis C virus (HCV) infection is unclear. OBJECTIVE: To compare histologic scoring of liver pathology in patients with chronic HCV infection with normal or elevated serum ALT. METHODS: Liver biopsies were performed in patients with HCV infection and either normal (n=40) or elevated (n=76) serum ALT levels, and scored for activity and fibrosis using the modified histological activity index. RESULTS: Patients with normal ALT and elevated ALT had similar demographic features. Median (range) histological activity grade was higher in patients with elevated ALT than in those with normal ALT (6 [1-15] vs. 5 [0-11], respectively; p=0.001), as was the fibrosis stage (2 [0-6] vs. 1[0-6]; p=0.02). Two patients with normal ALT and 4 with elevated ALT had liver cirrhosis. CONCLUSIONS: Among patients with chronic HCV infection, liver lesions are milder in those with normal serum ALT levels than those with abnormal ALT levels. However, some patients with normal ALT too may have advanced liver disease. PMID- 15879649 TI - Cholestatic jaundice during infancy: experience at a tertiary-care center in Bangladesh. AB - BACKGROUND AND OBJECTIVE: Cholestatic jaundice in early infancy is a difficult diagnostic problem. Early diagnosis is important for proper management. This retrospective study was conducted to find out the etiology and clinical profile of neonatal cholestatic disorders in Bangladesh. SETTING: Tertiary-care hospital in a developing country. METHODS: Clinical profile and cause of cholestatic illness were studied in 62 infants with cholestatic jaundice developing before three months of age and persisting for more than two weeks. RESULTS: Neonatal hepatitis (22; 35.5%--17 with TORCH, 5 with urinary infection), followed by biliary atresia (16; 25.8%) and idiopathic neonatal hepatitis (15; 24.2%), were the commonest causes of cholestasis. Mean age at presentation was 3.5 months. Ten (62.5%) of 16 biliary atresia cases were male and jaundice appeared before 14 days in 14 (87.5%) cases. CONCLUSIONS: Neonatal hepatitis, biliary atresia and idiopathic neonatal hepatitis were the common causes of neonatal cholestasis in infancy. Though cholestatic jaundice developed early, most of the cases presented late. PMID- 15879650 TI - Bile leaks following surgery for hepatic hydatid disease. AB - BACKGROUND: Conservative surgery (cyst evacuation and partial pericystectomy) for hydatid cysts of the liver is known to be safe but is often associated with bile leak and its sequelae. METHODS: Case records of 86 patients undergoing surgery for hydatid cysts of the liver at a tertiary-care center in northern India over a 14-year period were reviewed retrospectively. RESULTS: Sixteen (18%) patients had jaundice and 36 (42%) had a cyst-biliary communication detected at surgery. Biliary complications developed in 14 (16%) patients. Bile leaks and bilio cutaneous fistulae were observed in 11 (13%) patients; the fistula output was low (< 300 mL/day) in 8 of these. Three patients had localized intra-abdominal bile collections; all 3 underwent percutaneous drainage of biloma (subsequent laparotomy and lavage was required in one patient due to failure of percutaneous drainage), producing controlled low-output bilio-cutaneous fistulae in all. All low-output fistulae closed spontaneously after a mean duration of 4 weeks. Patients with high-output fistulae underwent endoscopic intervention (stenting/naso-biliary drainage), resulting in the conversion of these fistulae to low-output category and eventual closure after a mean duration of 7.5 weeks. CONCLUSION: Postoperative bile leaks lead to significant morbidity after surgical management of hydatid cysts of liver. A majority of them resolve spontaneously. Biliary drainage (endoscopic or surgical) hastens the closure of these bilio cutaneous fistulae. PMID- 15879651 TI - Peptic ulcer bleeding: is Helicobacter pylori a risk factor in an endemic area? AB - BACKGROUND/OBJECTIVE: A high prevalence of Helicobacter pylori infection has been reported in Iran. Although the importance of H. pylori in the induction of peptic ulcer disease is clearly defined, only few studies have addressed its role in bleeding from peptic ulcers. We evaluated the role of H. pylori in peptic ulcer bleeding. METHODS: Patients with acute peptic ulcer bleeding (PUB) and those with peptic ulcer disease without bleeding ('controls') were enrolled. Upper GI endoscopy and rapid urease test were performed in both groups. Histological study for detection of H. pylori was performed in patients with active bleeding, if RUT was negative. Other variables evaluated included sex, age, smoking, previous history of bleeding, non-steroidal anti-inflammatory drugs use, ulcer size, ulcer location, and duration of acid-peptic disease. Multivariate logistic regression analysis was performed to identify independent risk factors. RESULTS: 161 patients with PUB and 287 control patients were enrolled. H. pylori infection was seen more frequently in patients with duodenal ulcer than gastric ulcer (88.9% vs. 60.5%, p< 0.001). Univariate analysis showed that patients with PUB were more often male, older in age, used NSAID, had history of PUB in the past, had ulcer located in the stomach and not in the duodenum, and more often had large ulcer (>1 cm). Logistic regression analysis showed that H. pylori infection was protective in PUB after controlling for confounders (OR 0.41, 95% CI 0.21-0.79), when ulcer location was not entered in the model. A second model including ulcer location (to test for a residual effect) showed that H. pylori infection was not a significant risk factor in PUB (OR 0.61, 95% CI 0.30-1.24). CONCLUSIONS: H. pylori may not be an independent factor in bleeding from peptic ulcers. The lower frequency of this infection in these patients can be described by the higher frequency of bleeding from gastric ulcers, which are less H. pylori related compared with duodenal ulcer. PMID- 15879652 TI - Impact of intra-operative ultrasonography in liver surgery. AB - BACKGROUND/OBJECTIVE: Intra-operative ultra-sonography (IOUS) during surgery for primary and metastatic hepatic tumors identifies additional lesions and helps in determining the most optimal surgical strategy. We assessed the impact of IOUS in liver surgery at our hospital, a tertiary-care cancer center. METHODS: Patients with potentially resectable hepatic tumors underwent surgical exploration. The relationship of the tumor with regard to the intrahepatic vasculature was determined by IOUS. A search was also made for additional lesions not detected by pre-operative imaging modalities. In appropriate cases, IOUS was also used to assist resection and radiofrequency ablation/ethanol injection. RESULTS: Between January 2003 and January 2005, 52 patients underwent surgery for primary or secondary hepatic tumors. IOUS was performed in 48 of these patients. It detected additional hepatic lesions in 14 patients (29.2%). IOUS contributed to changing the operative plan in 21 patients (43.8%). It was directly responsible for avoiding resection or ablation in 7 patients (14.6%), 5 of whom had multiple bilobar lesions, 1 had IOUS-guided biopsy that revealed caseating granuloma on frozen section, and 1 patient had no lesion on IOUS. Three patients had extent of resection changed based on IOUS findings. IOUS also guided radiofrequency ablation in 8 patients and ethanol injection in one patient. CONCLUSION: IOUS is an essential tool in surgery for hepatic tumors. In addition to accurate staging, it also aids in safe resection and radiofrequency ablation in appropriate cases. PMID- 15879653 TI - Should acute viral C hepatitis be treated with interferon? PMID- 15879654 TI - Acute viral hepatitis C should be treated. PMID- 15879655 TI - Hepatitis C virus genotype 6 infection in India. AB - We report two patients with chronic liver disease--a 46-year-old man and a 52 year-old woman, both from eastern India--who were found to be infected with hepatitis C virus genotype 6 strains. These strains have been previously reported only from Hong Kong and Southeast Asia. PMID- 15879656 TI - Spontaneous antegrade enteral migration of feeding jejunostomy tube. AB - Spontaneous migration of enteral feeding tubes is unusual. We report an 18-year old man with corrosive stricture of the upper esophagus in whom feeding jejunostomy was performed using an 18F Levine's tube. Thirteen months later, he presented with absence of the tube. The patient was tolerating liquid diet and was managed conservatively. Serial radiographs and colonoscopy showed a steady progress of the tube through the gut. The tube was passed out spontaneously after 20 days. PMID- 15879657 TI - Pancreatic carcinoma: report of two cases presenting with unusual metastases. AB - We report two patients with adenocarcinoma of the body and tail of the pancreas who presented with features localized to the metastatic sites. A 67-year-old gentleman presented with right groin mass due to spermatic cord metastasis and later developed duodenal obstruction; the other gentleman was 69 years old and presented with change of bowel habit as a result of pelvic/pararectal metastasis. PMID- 15879658 TI - Ischemic stricture of Roux-en-Y intestinal loop and recurrent cholangitis. AB - The commonest complication of hepaticojejunostomy for the management of biliary strictures is recurrent cholangitis. We report a 54-year-old man who underwent choledochojejunostomy after choledochal cyst excision, and later developed ischemic stricture of the Roux-en-Y loop intestinal loop and recurrent cholangitis. The stricturous intestinal loop was excised with re-anastomosis with new Roux-en-Y loop, with uneventful recovery. PMID- 15879659 TI - Cholestatic liver injury due to ibuprofen. AB - Ibuprofen is a member of the propionic acid class of NSAID. We report a 35-year old man with ibuprofen-induced acute severe cholestatic liver injury. He recovered after seven months. PMID- 15879660 TI - Hyperhomocysteinemia presenting as superior mesenteric artery thrombosis. AB - We report a 23-year-old man who presented with acute abdomen. At laparotomy, he was diagnosed to have superior mesenteric artery thrombosis, with consequent extensive intestinal gangrene extending from the proximal jejunum till the mid transverse colon. He subsequently developed dry gangrene of the digits. Further evaluation showed that he had marked hyperhomocysteinemia. The gangrenous bowel was resected, and the homocysteine level normalized with folic acid supplementation. He is well at 1-year follow up. His brother, who was asymptomatic, was also detected to have hyperhomocysteinemia, which responded to folic acid. PMID- 15879661 TI - Inflammatory pseudotumor of gall bladder fossa of liver. AB - Inflammatory pseudotumor has been described in the lung, liver and other sites, but pseudotumors of the gall bladder fossa have not been reported earlier. We report a 39-year-old woman with inflammatory pseudotumor of the liver in the gall bladder fossa that resembled carcinoma gall bladder. PMID- 15879662 TI - Severe esophagitis in a child with Henoch-Schonlein purpura presenting as protein losing enteropathy. AB - A 4-year-old boy was seen for vomiting, diarrhea and peripheral edema. He had no evidence of nephrosis or liver dysfunction. Upper gastrointestinal endoscopy was performed for investigating the etiology of protein-losing enteropathy. It showed severe esophagitis and multiple ulcers in the descending duodenum. The symptoms and endoscopic mucosal abnormalities subsided after three weeks of hospitalization without specific therapy. Ten days after being discharged he was seen again with characteristic rash of Henoch-Schonlein purpura and arthritis without gastrointestinal symptoms. Biopsy of the skin rash revealed leukocytoclastic vasculitis. PMID- 15879663 TI - Hepatitis B vaccination status in health-care workers. PMID- 15879664 TI - Adult intussusception. PMID- 15879665 TI - Hepatitis C infection correlates with alteration of serum immunoglobulins pattern in chronic liver disease. PMID- 15879666 TI - Gastric cancer in southern Croatia during 2002-2003. PMID- 15879667 TI - Treatment of appendiceal mass. PMID- 15879668 TI - Prevalence of cholelithiasis in children--a hospital-based observation. PMID- 15879669 TI - Lymphoplasmacytic sclerosing pancreatitis mimicking pancreatic cancer. PMID- 15879670 TI - Henoch-Schonlein purpura probably due to montelukast presenting as subacute intestinal obstruction. PMID- 15879672 TI - Safe liver biopsy in a patient with chronic hepatitis C under continuous ambulatory peritoneal dialysis treatment. PMID- 15879673 TI - Spontaneous giant intramural hematoma of esophagus and stomach. PMID- 15879674 TI - Inhibitory action of 5-HT1A agonist MKC-242 on triazolam-induced phase advances in hamster circadian activity rhythms. AB - Mammalian circadian rhythms can be entrained by photic and non-photic stimuli. Although we know that non-photic entrainment interferes with photic entrainment signals, there is no information about whether photic entrainment interferes with non-photic entraining signals. We examined whether triazolam (TRZ), a non-photic form of entrainment, could be attenuated by pre-treatment with (S)-5-[3-[(1,4 benzodioxan-2-ylmethyl)amino]propoxy]-1,3-benzodioxole hydrochloride (MKC-242), a photic enhancing drug. We found that TRZ-induced phase advances in hamster behavioral circadian rhythms and the increase of locomotor activity were both significantly attenuated by MKC-242. Thus, in hamsters, the photic enhancing drug MKC-242 seemed to hinder benzodiazepine-induced non-photic entrainment. PMID- 15879675 TI - Abrogation of bronchial eosinophilic inflammation and attenuated eotaxin content in protease-activated receptor 2-deficient mice. AB - Protease-activated receptor 2 (PAR2) belongs to the PAR family (PAR1 to PAR4), which is activated by serine proteases (trypsin, tryptase, etc.). In this study, we evaluated the role of PAR2 in allergic inflammation of airways using PAR2 deficient (PAR2(-/-)) mice. In wild- type mice, infiltration of eosinophils and high eotaxin content were found in bronchoalveolar lavage fluid (BALF) after ovalbumin (OA) sensitization and following challenge. In contrast, both OA induced infiltration of eosinophils and increase of eotaxin content were abrogated in BALF from PAR2(-/-) mice. The activation of PAR2 might be essential in the production of eotaxin and consequential allergic inflammation in airways. PMID- 15879676 TI - Prostaglandin D2 inhibits IgE-mediated scratching by suppressing histamine release from mast cells. AB - Effects of prostaglandin (PG) D(2), PGE(2), and PGI(2) on itch-associated scratching responses of mice and histamine release from the rat basophilic leukemia cell line RBL-2H3 were examined. PGD(2) and ketotifen but not PGE(2) and PGI(2) suppressed the scratching caused by ovalbumin injected into ovalbumin sensitized mice. Ketotifen also suppressed compound 48/80-induced scratching but not PGD(2), PGE(2), and PGI(2). In vitro, PGD(2) suppressed the antigen-induced histamine release from RBL-2H3 cells, but PGE(2) and PGI(2) did not. These findings suggest that PGD(2) specifically suppressed IgE-mediated scratching by inhibiting IgE-mediated histamine release from mast cells. PMID- 15879677 TI - Cephaloridine induces translocation of protein kinase C delta into mitochondria and enhances mitochondrial generation of free radicals in the kidney cortex of rats causing renal dysfunction. AB - We have previously reported that the enhancement of free radical generation in mitochondria isolated from the kidney cortex of rats exposed to cephaloridine (CER) is probably mediated by the activation of protein kinase C (PKC). We examined which isoenzymes of PKC might be involved in the development of nephrotoxicity induced by CER in rats. The CER-induced renal dysfunction observed 24 h after its injection was prevented by a potent antioxidant DPPD and well known PKC inhibitors like H-7 and rottlerin. At 1.5 and 3.5 h after the CER injection, the free radical generation was increased markedly and this was associated with translocation of PKCdelta into the mitochondria of renal cortex tissue. Pretreatment of rats with H-7, a PKC inhibitor, significantly inhibited the CER-derived increase in mitochondrial generation of free radicals, suggesting that H-7 probably gets into the mitochondria and inhibits the activity of translocated PKC within the mitochondria. It was also shown that pretreatment of rats with rottlerin, a specific inhibitor of PKCdelta, suppressed the early translocation of PKCdelta into mitochondria and inhibited the CER-derived development of renal dysfunction. These results suggest that the CER-derived early translocation of PKCdelta into mitochondria probably leads to the enhanced production of free radicals through the mitochondrial respiratory chain during the development of the nephrotoxicity caused by CER. Understanding the role of PKCdelta in mitochondria may provide an important clue to the molecular mechanisms of mitochondrial production of reactive oxygen species and the free radical-induced renal failure in rats treated with CER. PMID- 15879678 TI - Pepsin-digested bovine lactoferrin induces apoptotic cell death with JNK/SAPK activation in oral cancer cells. AB - Lactoferrin, a member of the transferrin family, is iron-binding and a strongly cationic 76 kDa glycoprotein. In breast milk it is secreted in high concentrations from glandular epithelia and is also present in other exocrine fluids including saliva. In the present study, we examined the biological mechanisms of apoptosis induced by pepsin-digested-lactoferrin peptide (Lfn-p) in the human oral squamous cell carcinoma cell line SAS. We found that treatment with Lfn-p induced cell death with apoptotic nuclear changes, preceded by the cleavage of caspase-3 and poly (ADP-ribose) polymerase (PARP) in the apoptotic cells. Treatment with Lfn-p induced phosphorylation of extracellular signal regulated kinase (ERK1/2), a member of the MAP kinase family, at early stages of apoptosis. Another MAP kinase, c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK), was also phosphorylated by treatment with Lfn-p. Pretreatment of SAS cells with SP600125, a JNK/SAPK inhibitor, diminished Lfn-induced apoptosis, as assessed by determining released lactate dehydrogenase activity. On the other hand, the MEK1 inhibitors PD98059 or U0126 showed no effect on repression of cell death, but rather an increase. These results suggest that JNK/SAPK activation may play an important role in Lfn-p-induced apoptotic cell death of human oral squamous cell carcinoma cells. PMID- 15879679 TI - Atria selective prolongation by NIP-142, an antiarrhythmic agent, of refractory period and action potential duration in guinea pig myocardium. AB - NIP-142 is a novel benzopyran compound that was shown to prolong the atrial effective refractory period and terminate experimental atrial fibrillation in the dog. In the present study, we examined the effects of NIP-142 on isolated guinea pig myocardium and on the G-protein-coupled inwardly rectifying potassium channel current (acetylcholine-activated potassium current; I(KACh)) expressed in Xenopus oocytes. NIP-142 (10 and 100 microM) concentration-dependently prolonged the refractory period and action potential duration in the atrium but not in the ventricle. E-4031 and 4-aminopyridine prolonged action potential duration in both left atrium and right ventricle. Prolongation by NIP-142 of the atrial action potential duration was observed at stimulation frequencies between 0.5 and 5 Hz. In contrast, the prolongation by E-4031 was not observed at higher frequencies. Tertiapin, a blocker of I(KACh), prolonged action potential duration in the atrium but not in the ventricle. NIP-142 completely reversed the carbachol induced shortening of atrial action potential duration. NIP-142 (1 to 100 microM), as well as tertiapin (0.1 to 100 nM), concentration-dependently blocked I(KACh) expressed in Xenopus oocytes; the blockade by NIP-142 was not affected by membrane voltage. In conclusion, NIP-142 was shown to prolong atrial refractory period and action potential duration through blockade of I(KACh) which may possibly explain its previously described antiarrhythmic activity. NIP-142 has pharmacological properties that are different from classical class III antiarrhythmic agents such as atria specificity and lack of reverse frequency dependence, and thus appears promising for the treatment of supraventricular arrhythmia. PMID- 15879680 TI - Modification of kappa-opioid receptor agonist-induced antinociception by diabetes in the mouse brain and spinal cord. AB - The supraspinal and spinal antinociceptive effects of several kappa-opioid receptor agonists were examined in diabetic and non-diabetic mice using the tail flick assay. The antinociception induced by intrathecal (i.t.), but not intracerebroventricular (i.c.v.), CI-977, a highly selective kappa(1)-opioid receptor agonist, in diabetic mice was less than that in non-diabetic mice. The antinociceptive effects of ICI-199,441 and R-84760, high potency kappa(1)-opioid receptor agonists, given i.c.v., but not i.t., were attenuated in diabetic mice compared to those in non-diabetic mice. On the other hand, the antinociceptive effects of the new kappa-opioid receptor agonist TRK-820, which has high affinity for kappa(2)- and/or kappa(3)-opioid receptors, injected both i.c.v. and i.t. in diabetic mice were markedly less than those in non-diabetic mice. These results indicate that the antinociceptive effects of those kappa-opioid receptor agonists in diabetic mice are altered in a region-specific manner in the central nervous system (CNS). The dysfunction of kappa-opioid receptor subtypes in diabetic mice may underlie this CNS region-specific variation in the effects of these kappa opioid receptor agonists. PMID- 15879681 TI - (+)-Catechin, an ingredient of green tea, protects murine microglia from oxidative stress-induced DNA damage and cell cycle arrest. AB - Extracts of Chinese herbs have been demonstrated to inhibit oxidative stress in vitro. In this study, we investigated the mechanism of (+)-catechin, isolated from green tea, which preserved murine microglia N9 cells from an oxidative agent tert-butylhydroperoxide (tBHP)-induced cell death. (+)-Catechin augmented the cell survival ratio after exposure to tBHP. Protective action of this drug was more efficacious than that of N-acetylcysteine, which is a putative antioxidant. DNA damage, detected by the Comet assay, was diminished with treatment of the drug. Results of flow cytometric analysis showed that the amount of intracellular *OH was decreased, and the cell cycle arrest was reversed by down-regulation of p53 phosphorylation after treatment with (+)-catechin. The reduced p53 activity followed the impairment of NF-kappaB translocation to the nuclear region. Then the phosphorylation of extracellular signal regulated protein kinase, a cell survival facilitative signal, was upregulated at the later stage. Taken together, (+)-catechin inhibited tBHP-induced translocation of NF-kappaB to improve cellular survival. PMID- 15879682 TI - Shengmai San, a Chinese herbal medicine protects against rat heat stroke by reducing inflammatory cytokines and nitric oxide formation. AB - The aim of the present study was to ascertain whether the possible occurrence of overproduction of inducible nitric oxide synthase (iNOS)-dependent nitric oxide (NO) in the brain and inflammatory cytokines in the peripheral blood exhibited during heat stroke can be reduced by prior administration of Shengmai San, a Chinese herbal medicine. Aminoguanidine, an iNOS inhibitor, was evaluated at the same time as a reference (positive control). Urethane-anesthetized rats were exposed to heat stress (ambient temperature of 43 degrees C) to induce heat stroke. Control rats were exposed to 24 degrees C. Mean arterial pressure and cerebral blood flow after the onset of heat stroke were all significantly lower than in control rats. However, cerebral iNOS immunoreactivity and NO levels were all greater after the onset of heat stroke. The serum levels of interleukin 1beta, interleukin-6, and tumor necrosis factor-alpha were all increased after the onset of heat stroke. Shengmai San (1.2 g/ml per rat) or aminoguanidine (30 micromol/ml per rat) was administered orally, daily, and consecutively for 7 days before the initiation of heat stress; and this significantly attenuated the heat stress-induced arterial hypotension, cerebral ischemia, and increased levels of brain iNOS-dependent NO production and serum cytokines formation. Shengmai San shared with the aminoguanidine almost the same efficacy in reducing iNOS dependent NO and cytokines overproduction during heat stroke. These results suggest that Shengmai San or aminoguanidine protects against heat stroke-induced arterial hypotension and cerebral ischemia by inhibition of iNOS-dependent NO overproduction in the brain and excessive accumulation of several inflammatory cytokines in the peripheral blood stream. PMID- 15879683 TI - The IGF system: new developments relevant to pediatric practice. AB - In the 50 years since the initial report of the 'sulfation factor' or 'somatomedin'hypothesis, the IGF system has established itself as the primary mediator of both intrauterine and postnatal growth in mammals. IGF deficiency (IGFD) has emerged as an important clinical diagnosis: secondary IGFD results from insufficient production of GH and is characterized by postnatal growth failure; primary IGFD can result from abnormalities of the GH receptor or GH signaling cascade, or from mutations or deletions of the IGF-I gene. Monitoring IGF production during short-term IGF generation tests or during chronic GH therapy can provide a means for evaluating the efficacy and safety of GH treatment. IGF-I, either alone or in combination with IGF binding proteins, is the treatment of choice for primary IGFD and may have a role in treatment of idiopathic short stature when accompanied by decreased serum concentrations of IGF-I. PMID- 15879684 TI - Clinical relevance of systemic and local IGF-I. AB - The insulin-like growth factor family of ligands, receptors and binding proteins are critical for many normal physiological functions. These include normal development during fetal and post-natal development and maintenance of organ function in adult life. Circulating IGF-I is produced primarily by the liver under GH control, whereas the production of tissue IGF-I has other controls. Recent studies have demonstrated that both circulating and tissue IGF-I are important for maintaining the normal structure-function of complex organs such as bone. Circulating IGF-I is important for maintaining ambient GH levels; in its absence GH elevation is seen leading to insulin resistance. In addition, low levels of circulating IGF-I retard the progression and metastatic potential of a number of cancers. PMID- 15879685 TI - Cartilage disorders: potential therapeutic use of mesenchymal stem cells. AB - Chondrogenesis is a well-orchestrated process driven by chondroprogenitors that undergo to condensation, proliferation and chondrocyte differentiation. Because cartilage lacks regenerative ability, treatments for cartilage diseases are primarily palliative. Adult bone marrow contains a reservoir of mesenchymal stem cells (MSC) with in vitro and in vivo potential of becoming cartilage. To optimize the potential therapeutic use of MSC in cartilage disorders, we need to understand the mechanisms by which growth factors determine their chondrogenic potential. Insulin-like growth factors (IGFs) play a central role in chondrogenesis as indicated by the severe growth failure observed in animals carrying null mutations of Igfs and Igf1R genes. We have found that IGF-I has potent chondrogenic effects in MSC. Effects are similar to transforming growth factor-Beta (TGF-Beta). Insulin-like growth factor binding protein-3 (IGFBP-3), well characterized as IGF carrier, has intrinsic bioactivities that are independent of IGF binding. IGFBP-3 levels are increased in degenerative cartilage diseases such as osteoarthritis. We have demonstrated that IGFBP-3 has IGF-independent growth inhibitory effects in chondroprogenitors. We now show that IGFBP-3 induces MSC apoptosis and antagonizes TGF-Beta chondroinductive effects in chondroprogenitors. Understanding IGF-I chondroinductive and IGFBP-3 chondroinhibitory effects would provide critical information to optimize the therapeutic use of MSC in cartilage disorders. PMID- 15879686 TI - Treatment in animal models. AB - It is now well established that neurons and other cell types may die many hours or even days after hypoxic-ischemic injury due to activation of programmed cell death (apoptotic) pathways. The potent anti-apoptotic factor IGF-1 and its binding proteins and receptors are intensely induced within damaged brain regions following brain injury suggesting a possible a role for IGF-1 in endogenous brain recovery. Exogenous administration of IGF-1 within a few hours after brain injury has now been shown to be protective in both grey and white matter, and leads to improved long-term neurological function. The limited window of opportunity for treatment with IGF-1 can be extended by spontaneous mild post-hypoxic hypothermia, probably due to delayed evolution of apoptotic processes. The efficacy of IGF-1 is specific to particular cellular phenotypes and brain regions, and its neuroprotective effects are mediated by IGF-1 receptors and binding proteins. Intriguingly its naturally cleaved N-terminal tripeptide (glycine-proline-glutamate, GPE) has been demonstrated to be neuroprotective after both central and peripheral administration. Peripheral administration of GPE also prevents the loss of dopamine neurons and improves long-term functional recovery following 6-OHDA lesion. However, GPE is unlikely to contribute significantly to the direct effects of IGF-1. PMID- 15879687 TI - Insulin-like growth factor-I and risk of type 2 diabetes and coronary heart disease: molecular epidemiology. AB - Insulin-like growth factors (IGFs) play a fundamental role in somatic growth and cellular differentiation, metabolism and survival. Indeed, the processes linking nutrition, metabolism and growth are thought to involve a complex interrelation among insulin, growth hormone (GH), IGFs and their binding proteins (IGFBPs). However, accumulating data from both experimental and molecular epidemiological studies indicate that these growth factors may also be important in the pathophysiological processes underlying chronic disease, including type 2 diabetes mellitus, coronary heart disease and cancer. Experimental and observational studies suggest that higher levels of circulating IGF-I may increase risk of several cancers. By contrast, recent prospective epidemiological studies suggest that relatively higher IGF-I levels may reduce the risk of type 2 diabetes and coronary heart disease. However, these relatively small-scale observational studies are susceptible to chance, reverse causality and residual or unmeasured confounding. A 'Mendelian randomization' approach based on large scale gene association and prospective observational studies might help determine the possible causal role of IGF-I and its binding proteins in the aetiology of type 2 diabetes, coronary heart disease and cancer. PMID- 15879688 TI - Quantitative measurement of IGF-I and its use in diagnosing and monitoring treatment of disorders of growth hormone secretion. AB - Quantitative measurements of serum IGF-I concentrations are extremely useful in the diagnosis of growth hormone deficiency in children and young adults and in the diagnosis of acromegaly. Similarly, IGF-I measurements have been of great use to clinical investigators who are monitoring the effect of either growth hormone or IGF-I in various disorders. Several guidelines have proven very useful for assaying IGF-I and defining high-quality assays. Since IGF-I circulates associated with IGF binding proteins it is essential that a high-quality assay remove binding protein interference. Similarly, it is essential that the antibodies that are utilized in the assay have both high specificity and high affinity for IGF-I. Assay reproducibility is effected by such factors as quality of sample collection, utilization of proper internal and external standards and published validation using not only normal samples but samples from a variety of pathophysiologic conditions. Interlaboratory variability often is due to differences in one of more of these characteristics. When assessing the validity of abnormal assay results it is important to be able to compare the value to a normative dataset that has been developed on a large number of subjects. Recently a normative dataset was published on 2,812 subjects and this provides an important reference standard by which other assays should be judged. Careful attention to these characteristics is likely to result in valid and useful IGF-I assay results. PMID- 15879689 TI - IGF-I and IGFBP-3 assessment in the management of childhood onset growth hormone deficiency. AB - The diagnosis of growth hormone (GH) deficiency (GHD) in childhood is not straightforward, being still based on a comprehensive clinical, anthropometric, endocrine and neuroradiological assessment. Due to their GH dependency and relative stability in circulation, IGF-I and IGFBP-3 serum concentrations were proposed as reliable indicators of daily GH secretion. However, the sensitivity of assays for both IGF-I and IGFBP-3 is inadequate to exclude the diagnosis of GHD merely on the basis of a normal value of the two parameters, although it seems likely that IGF-I values higher than -1 SD reflect a normal GH secretion. On the other hand, as the specificity of both measurements is over 90%, subnormal concentrations strongly support the diagnosis of GHD. Finally, combining the evaluation of growth velocity with IGF-I measurement, sensitivity and specificity reach a value > or =95%, implying that two subnormal values strongly suggest and two normal values strongly oppose the diagnosis of GHD. PMID- 15879690 TI - IGFs and IGFBPs in adult growth hormone deficiency. AB - In the current guidelines for the diagnosis of adult GH deficiency (GHD) it is stated that, within the appropriate clinical context, it has to be shown by provocative tests only. But the diagnostic value of measuring IGF-I levels has been recently revisited. It has been confirmed that normal IGF-I levels do not rule out severe GHD in adults. However, it has also been emphasized that very low IGF-I levels in patients highly suspected for GHD (and without malnutrition, liver disease or hypothyroidism) could be considered definite evidence for severe GHD. This assumption particularly applies to patients with childhood-onset, severe GHD or with multiple hypopituitarism acquired in adulthood. The value of measuring IGF-I levels for monitoring the efficacy and the adequacy of rhGH replacement remains definitely accepted. PMID- 15879691 TI - Monitoring of insulin-like growth factors during growth hormone treatment: adulthood growth hormone deficiency. AB - Decisions on growth hormone (GH) dosing in paediatric practice have depended on body weight or surface area calculations. Although this policy was used initially in the treatment of adult patients, it has become apparent both in clinical studies and longitudinal observation that this results in relative overtreatment of male patients, with a high incidence of immediate side effects, and undertreatment of females. Use of individualized GH dosing against serum IGF-I has proven useful in optimizing therapy across the whole adult age range, including the elderly and in patients who are in transition from post-puberty to full somatic development. It also eliminates gender differences in susceptibility. In this respect, serum IGF-I is superior to other GH-dependent peptides. However, on present evidence, serum IGF-I should be regarded primarily as a safety marker rather than a specific therapeutic marker. PMID- 15879692 TI - IGFs and IGFBPs in GH insensitivity. AB - IGF-I, IGFBP-3 and ALS are GH-dependent peptides and their production is disturbed in states of GH insensitivity. This chapter explores the relative degrees of IGF-I, IGFBP-3 and ALS deficiency across the spectrum of GH insensitivity. In classical GH insensitivity syndrome (GHIS), known as Laron syndrome, due to GH receptor (GHR) deficiency, serum IGF-I, IGFBP-3 and ALS are severely reduced with inability to produce these peptides during an IGF-I generation test. Across the spectrum of severity of GHR defects, some patients have short stature and normal facial appearance, so-called partial or non classical GH insensitivity. In these cases the IGF-I, IGFBP-3 deficiency is less severe. A positive relationship exists between height SDS and IGFBP-3 SDS (r2 = 0.45, p < 0.001) in patients from the European series with GHIS. In a new series of GHIS cases (n = 36) there was a significant difference in IGFBP-3 and ALS (p < 0.05) between classical (n = 25) and non-classical cases (n = 11). IGF-I, IGFBP-3 and ALS were significantly higher (p < 0.05) in pubertal compared with pre pubertal subjects in the same series. In idiopathic short stature (ISS), heterozygous mutations of the GHR may have a dominant negative effect. ISS patients have lower IGF-I levels than the normal population. In 21 cases, mean IGF-I SDS was -1.39 (-2.4 to -1.16) and IGFBP-3; -0.45 (-1.13 to 0.38). However, IGF-I and IGFBP-3 responses in the IGF-I generation test were generally normal. In acquired GHI due to chronic illness such as Crohn's disease, juvenile arthritis and cystic fibrosis, IGF-I deficiency is present, although IGFBP-3 is usually normal. In summary, assessment of IGF-I, IGFBP-3 and ALS contributes to diagnosis in GH insensitivity states. In our experience, IGF-I is more sensitive to disturbance of GH action that IGFBP-3, however in severe GHIS cases, IGF-I is usually undetectable and measurement of IGFBP-3 is valuable as a guide to the severity of the biological defect. PMID- 15879693 TI - Childhood and adolescent diabetes. AB - Circulating levels of insulin-like growth factor-I (IGF-I) and its principal binding protein IGFBP-3 are reduced, whereas those of the inhibitory binding protein, IGFBP-1, tend to be high in children and adolescents with type 1 diabetes mellitus (T1DM). These abnormalities are thought to arise because of relative portal hypoinsulinaemia and partial resistance at the hepatic growth hormone (GH) receptor. During adolescence, reductions in IGF-I and IGF bioactivity lead to feedback for GH hypersecretion and the elevated GH and low IGF-I levels lead to an increase of the normal insulin resistance encountered during puberty. Low IGF-I levels, but in particular elevated GH levels, have been implicated in the pathogenesis of diabetic microangiopathic complications, in particular, renal hypertrophy, glomerular hyperfiltration and the development of microalbuminuria. Early study of IGF-I replacement with recombinant human IGF-I (rhIGF-I) demonstrated, in the short term, reductions in GH hypersecretion with improved insulin sensitivity and, in the longer term, reductions in insulin requirements and improvements in HbA1c levels. However, larger doses of rhIGF-I were associated with retinopathy either due to rapid improvements in glycaemic control or direct effects of high levels of 'free' IGF-I. More recently, pilot studies using the combination of rhIGF-I/rhIGFBP-3 have confirmed the physiological efficacy of IGF-I replacement in T1DM. The combined treatment is better tolerated and may result in reduced tissue exposure to high levels of 'free' IGF-I. Longer term clinical studies with this IGF-I/IGFBP-3 combination are needed. PMID- 15879694 TI - Insulin resistance and type 2 diabetes mellitus: is there a therapeutic role for IGF-1? AB - Severe insulin resistance represents a heterogeneous set of conditions with diverse underlying pathophysiology and limited therapeutic options. While recent studies have identified some of the molecular mechanisms resulting in extreme insulin resistance in peripheral tissues and liver, these studies have resulted in relatively few advances in therapy. Insulin like-growth factor 1 (IGF-1) lowers blood glucose while at the same time lowering serum insulin levels in normal volunteers. Its mechanism of action appears to be independent of activation of the insulin receptor although the role of IGF-1 in normal carbohydrate metabolism remains incompletely defined. IGF-1 also improves insulin resistance both in type 2 diabetes and in subjects with more severe insulin resistance. Small-scale clinical trials have demonstrated the potential utility of rhIGF-1 in selected cases of severe insulin resistance and, in these cases, the risk-benefit ratio appears to favor the use of this drug to ameliorate biochemical abnormalities and clinical symptoms. PMID- 15879695 TI - Insulin-like growth factors in the treatment of neurological disease. AB - Although the functional deficits of neurological diseases vary, they are all pathologically marked by neuronal degeneration. The ability of insulin-like growth factor-I (IGF-I) to support both sensory and motor neuron regeneration has suggested its potential in treatment of neurological diseases. IGF-I is pleiotrophic, stimulating survival, neurite outgrowth and motility in neurons, as well as myelination of neurons by glia. Understanding the intracellular signaling pathways that mediate these pleiotrophic responses to IGF-I is important for tailoring IGF-I treatment to the appropriate neurological deficit. This review surveys the current understanding of IGF-I pleiotrophism, the underlying signaling conferring these effects, and the status of IGF-I in treatment of human neurological disorders. PMID- 15879696 TI - Insulin-like growth factor system in amyotrophic lateral sclerosis. AB - Insulin-like growth factor-I (IGF-I) is a neurotrophic factor with insulin-like metabolic activities, and possesses potential clinical applications, particularly in neurodegenerative disorders. Amyotrophic lateral sclerosis (ALS) is a chronic progressive devastating disorder of the central nervous system, characterized by the death of upper and lower motor neurons. Both in vivo and in vitro studies have shown that IGF-I promotes motor neuron survival and strongly enhances motor nerve regeneration. Evidence that IGF-I rescues motor neurons has led to clinical trials of human recombinant IGF-I in ALS patients. However, systemic delivery of human recombinant IGF-I in these trials did not lead to beneficial clinical effects in ALS patients and may be due through inactivation of IGF-I by binding to IGF binding proteins (IGFBPs), and or limited delivery of IGF-I to motor neurons. Recently it was shown that both IGF-I receptors and IGFBPs were increased on motor neurons of ALS patients and free levels of IGF-I were decreased by 50%. In this study it was suggested that IGFBPs inactivate IGF-I by forming inactive complexes. The uses of IGF analogues with low affinity for IGFBPs and analogues that are able to displace IGF-I from IGFBPs are better candidates in new clinical trials. Another possibility is to find a way of IGF-I transport without hindrance of circulating and tissue-specific IGFBPs, such as IGF-I delivery based on gene therapy. PMID- 15879697 TI - Recent progress of structural biology of tRNA processing and modification. AB - Transfer RNA (tRNA) is a key molecule to decode the genetic information on mRNA to amino aicds (protein), in a ribosome. For tRNA to fulfill its adopter function, tRNA should be processed into the standard length, and be post transcriptionally modified. This modification step is essential for the tRNA to maintain the canonical L-shaped structure, which is required for the decoding function of tRNA. Otherwise, it has recently been proposed that modification procedure itself contributes to the RNA (re)folding, where the modification enzymes function as a kind of RNA chaperones. Recent genome analyses and post genome (proteomics and transcriptomics) analyses have identified genes involved in the tRNA processings and modifications. Furthermore, post-genomic structural analysis has elucidated the structural basis for the tRNA maturation mechanism. In this paper, the recent progress of the structural biology of the tRNA processing and modification is reviewed. PMID- 15879698 TI - Drosophila melanogaster: a model for the study of DNA damage checkpoint response. AB - The cells of metazoans respond to DNA damage by either arresting their cell cycle in order to repair the DNA, or by undergoing apoptosis. This response is highly conserved across species, and many of the genes involved in this DNA damage response have been shown to be inactivated in human cancers. This suggests the importance of DNA damage response with regard to the prevention of cancer. The DNA damage checkpoint responses vary greatly depending on the developmental context, cell type, gene expression profile, and the degree and nature of the DNA lesions. More valuable information can be obtained from studies utilizing whole organisms in which the molecular basis of development has been well established, such as Drosophila. Since the discovery of the Drosophila p53 orthologue, various aspects of DNA damage responses have been studied in Drosophila. In this review, I will summarize the current knowledge on the DNA damage checkpoint response in Drosophila. With the ease of genetic, cellular, and cytological approaches, Drosophila will become an increasingly valuable model organism for the study of mechanisms inherent to cancer formation associated with defects in the DNA damage pathway. PMID- 15879699 TI - Expression of IL-17 homologs and their receptors in the synovial cells of rheumatoid arthritis patients. AB - IL-17 is a major proinflammatory cytokine secreted by activated T-lymphocytes that accumulates in the inflamed joints of rheumatoid arthritis (RA) patients. Additional IL-17-related molecules and their receptors have been discovered and may also contribute to RA pathogenesis. We examined the expression of the prototypic IL-17 (IL-17A) and its homologs, IL-17B-F, by RT-PCR analyses of synovial fluid mononuclear cells (SFMCs) and peripheral blood mononuclear cells (PBMCs) from RA patients. We also tested for induction of the IL-17 receptor homologs upon stimulation of the fibroblast-like synoviocytes (FLSs) of RA patients with IL-17. The patients' SFMCs expressed IL-17C, E and F in addition to IL-17A. As in the case of IL-17, IL-15 appears to be the major inducer of these homologs in RA SFMCs. We detected transcripts of IL-17R, as well as those of IL 17RB, C and D, in the FLSs of RA patients. Whereas IL-17R expression increased upon in vitro stimulation with IL-17, expression of IL-17RB, C and D was unchanged. However the possibility of cross-interaction between other IL-17 homologs and receptor isoforms remains to be investigated. Our data suggest that these additional homologs should also be considered as targets for immune modulation in the treatment of RA joint inflammation. PMID- 15879700 TI - Induction of apoptosis in chicken oviduct cells by C2-ceramide. AB - The chicken oviduct is a dynamic organ that produces secretory proteins such as ovalbumin and its cells undergo cell proliferation and differentiation. There has been no study of the cellular mechanism involved in cell death in the chicken oviduct. Therefore, this study has focused on the study of apoptosis in primary oviduct cells. Because ceramide is known to activate apoptosis in tumor cells and is produced in the oviduct, we used an exogenous ceramide analog to induce cell death. The viability of ceramide-treated chicken oviduct cells decreased in a dose-dependent manner and apoptotic cells were detected by staining with annexin V. The expression of apoptosis-related genes was assessed by RT-PCR and bcl-2 mRNA was found to decrease after exposure to ceramide while Bcl-x mRNA increased 12 h post-treatment. In addition, caspase-3 was expressed strongly in the early stages of apoptosis, while caspase-1 and -9 transcripts increased at later times. We conclude that ceramide induces apoptosis in oviduct-derived primary cells via a caspase- and bcl-2- dependent pathway. PMID- 15879701 TI - Enhanced transduction of Cu,Zn-superoxide dismutase with HIV-1 Tat protein transduction domains at both termini. AB - The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species. PMID- 15879702 TI - 3'-Half of the thrombopoietin cDNA confers higher expression of erythropoietin at the RNA level but not at the protein level. AB - Both erythropoietin (EPO) and the short-form thrombopoietin (TPO) were expressed at low levels whereas the long-form TPO was expressed at high levels in transgenic animals. To elucidate the role of carboxy-terminal half of the long form TPO which is absent in the short-form, we generated recombinant TPO or EPO expression vectors which contain or lack the carboxy-terminal half of TPO and examined their expression in the HC11 and 293 cells. The long-form TPO was expressed higher than the short-form regardless of the cell types, transfection modes, and promoters. When 3'-half of the long-form TPO cDNA was placed downstream of the EPO cDNA to act as a 3'-untranslated region, expression of EPO was moderately increased at the RNA level, however, no remarkable increase was observed at the protein level. These results suggest that the low expression of EPO, as like as the short-form TPO, is due to absence of the 3'-half in the full length TPO that confers stability both at the RNA and protein levels. PMID- 15879703 TI - Structure and function of RGD peptides derived from disintegrin proteins. AB - The Arg-Gly-Asp (RGD) sequence serves as the primary recognition site in extracellular matrix proteins, and peptides containing this sequence can mimic the biological activities of matrix proteins. We have initiated structure function studies of two RGD containing peptides, RGD-5(AGGDD) and cyclic RGD 6(CARGDDC). Assays have shown that cyclic RGD-peptides inhibit platelet aggregation more efficiently than linear ones. NMR data revealed that RGD-5 and RGD-6 have entirely different conformation. RGD-5 has a linear extended structure and RGD-6 has a stable loop conformation. In RGD-5 the guanidinium group of Arg2 and the carboxyl group of Asp4 lie in parallel, whereas the side-chains of Arg3 and Asp5 of RGD-6 are located in different planes, supporting the idea that the stability of the cyclic form derives from the packing of the side chain of the Arg and Asp residues. The structural features of these peptides could provide a basis for designing new drugs against diseases related to platelet aggregation and as cancer antagonists. PMID- 15879704 TI - Characterization of an abiotic stress-inducible dehydrin gene, OsDhn1, in rice (Oryza sativa L.). AB - A full-length 1.1 kb cDNA, designated Oryza sativa Dehydrin 1 (OsDhn1), was isolated from the seed coat of rice. The deduced protein is hydrophilic and has three K-type and one S-type motifs (SK3-type), indicating that OsDhn1 belongs to the acidic dehydrin family, which includes wheat WCOR410 and Arabidopsis COR47. Expression of OsDhn1 was strongly induced by low temperature as well as by drought. Induction of OsDhn1 by cold stress was clearcut in the roots of seedlings and the epidermis of palea and lemma. OsDhn1 was also up-regulated in UBI::CBF1/DREB1b transgenic plants indicating that it is regulated by the CBF/DREB stress signaling pathway. PMID- 15879705 TI - Crystallization and X-ray crystallographic studies of wild-type and mutant tryptophan synthase alpha-subunits from Escherichia coli. AB - The alpha-subunit of Escherichia coli tryptophan synthase (aTS), a component of the tryptophan synthase alpha2beta2 complex, is a monomeric 268-residues protein (Mr = 28,600). alphaTS by itself catalyzes the cleavage of indole-3-glycerol phosphate to glyceraldehyde-3-phosphate and indole, which is converted to tryptophan in tryptophan biosynthesis. Wild-type and P28L/Y173F double mutant alpha-subunits were overexpressed in E. coli and crystallized at 298 K by the hanging-drop vapor-diffusion method. X-ray diffraction data were collected to 2.5 angstroms resolution from the wild-type crystals and to 1.8 angstroms from the crystals of the double mutant, since the latter produced better quality diffraction data. The wild-type crystals belonged to the monoclinic space group C2 (a = 155.64 angstroms, b = 44.54 angstroms, c = 71.53 angstroms and beta = 96.39 degrees) and the P28L/Y173F crystals to the monoclinic space group P21 (a = 71.09 angstroms, b = 52.70, c = 71.52 angstroms, and beta = 91.49 degrees). The asymmetric unit of both structures contained two molecules of aTS. Crystal volume per protein mass (V(m)) and solvent content were 2.15 angstroms3 Da(-1) and 42.95% for the wild-type and 2.34 angstroms3 Da(-1) and 47.52% for the double mutant. PMID- 15879706 TI - Cloning and characterization of TMPRSS6, a novel type 2 transmembrane serine protease. AB - We have identified TMPRSS6, a novel type 2 transmembrane serine protease. TMPRSS6 possesses all the signature motifs of the family of transmembrane serine proteases (TMPRSSs), including a transmembrane domain, an LDL receptor class A (LDLRA) domain, a scavenger receptor cysteine-rich (SRCR) domain, and a serine protease domain. The substrate specificity of TMPRSS6 is slightly different from those of other TMPRSS family members. Combined with the finding that TMPRSS6 is expressed strongly in the thyroid and weakly in the trachea, this may indicate that TMPRSS6 has a specialized role. PMID- 15879707 TI - The tobacco ubiquitin-activating enzymes NtE1A and NtE1B are induced by tobacco mosaic virus, wounding and stress hormones. AB - Recent characterization of several genes involved in plant defense responses suggested that ubiquitin-mediated protein degradation has a role in these responses. We isolated two cDNAs (NtUBA1 and NtUBA2) encoding ubiquitin activating enzyme (E1) from Nicotiana tabacum cv. BY-2. The open reading frames of both encoded 1080 amino acids, corresponding to molecular masses of 120 kDa. The E1s and corresponding transcripts were upregulated by infection with tobacco mosaic virus (TMV) and tomato mosaic virus (ToMV), and to a lesser extent by cucumber mosaic virus (CMV). Furthermore, they were also upregulated by wounding stress, and the plant hormones salicylic acid, jasmonic acid and the ethylene precursor, aminocyclopropane-1-carboxylic acid (ACC). Our findings support the idea that the ubiquitin-proteasome system plays a role in plant disease defenses. PMID- 15879708 TI - Attenuation of extracellular acidic pH-induced cyclooxygenase-2 expression by nitric oxide. AB - Corneal endothelial cells play an important role in maintaining the transparency and ionic balance of the cornea. Inflammation causes many changes in the intracellular and extracellular environment of the cornea, including acidosis. We examined the relationship between changes in extracellular pH and expression of cyclooxygenase-2 in cultured bovine corneal endothelial cells. When extracellular pH ([pH]o) was reduced to pH 6.4, COX-2 mRNA increased, with a peak at 2 h. This was blocked by pretreatment with actinomycin D and incubation with spermine NONOate (SPER/NO, a nitric oxide donor). Exposure to the H+ ionophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), also raised COX-2 mRNA levels. CCCP induced COX-2 mRNA expression was also reduced by SPER/NO. These results were confirmed immuno-cytochemically. These data demonstrate that COX-2 expression is stimulated by the lowering of extracellular pH that could result from bacterial infection, and that this is countered by over-production of nitric oxide, which could also result from bacterial infection. PMID- 15879709 TI - Effects of FIS protein on rnpB transcription in Escherichia coli. AB - Factor for inversion stimulation (FIS), the Escherichia coli protein, is a positive regulator of the transcription of genes that encode stable RNA species, such as rRNA and tRNA. Transcription of the rnpB gene encoding M1 RNA, the catalytic subunit of E. coli RNase P, rapidly declines under stringent conditions, as does that of other stable RNAs. There are multiple putative FIS binding sites upstream of the rnpB promoter. We tested whether FIS binds to these sites, and if so, how it affects rnpB transcription. In vitro binding assays revealed specific binding of FIS to multiple sites in the rnpB promoter region. Interestingly, FIS bound not only to the upstream region of the promoter, but also to the region from +4 to +18. FIS activated rnpB transcription in vitro, but the level of activation was much lower than that of the rrnB promoter for rRNA. We also examined the effects of FIS on rnpB transcription in vivo using isogenic fis+ and fis- strains. rnpB transcription was higher in the fis- than the fis+ cells during the transitions from lag to exponential phase, and from exponential to stationary phase. PMID- 15879710 TI - Characterization of the functional domains of human foamy virus integrase using chimeric integrases. AB - Retroviral integrases insert viral DNA into target DNA. In this process they recognize their own DNA specifically via functional domains. In order to analyze these functional domains, we constructed six chimeric integrases by swapping domains between HIV-1 and HFV integrases, and two point mutants of HFV integrase. Chimeric integrases with the central domain of HIV-1 integrase had strand transfer and disintegration activities, in agreement with the idea that the central domain determines viral DNA specificity and has catalytic activity. On the other hand, chimeric integrases with the central domain of HFV integrase did not have any enzymatic activity apart from FFH that had weak disintegration activity, suggesting that the central domain of HFV integrase was defective catalytically or structurally. However, these inactive chimeras were efficiently complemented by the point mutants (D164A and E200A) of HFV integrase, indicating that the central domain of HFV integrase possesses potential enzymatic activity but is not able to recognize viral or target DNA without the help of its homologous N-terminal and C-terminal domains. PMID- 15879711 TI - Identification of a glucokinase that generates a major glucose phosphorylation activity in the cyanobacterium Synechocystis sp. PCC 6803. AB - In silico analysis of genome of the cyanobacterium Synechocystis sp. PCC 6803 identified two genes, slr0329 and sll0593, that might participate in glucose (Glc) phosphorylation (www.kazusa.or.jp/cyano). In order to determine the functions of these two genes, we generated deletion mutants, and analyzed their phenotypes and enzymatic activities. In the presence of 10 mM Glc, wild-type (WT) and slr0329 defective strain (M1) grew fast with increased respiratory activity and NADPH production, whereas the sll0593 deletion mutant (M2) failed to show any of the Glc responses. WT and M1 were not significantly different in their glucokinase activity, but M2 had 90% less activity. Therefore, we propose that Sll0593 plays a major role in the phosphorylation of glucose in Synechocystis cells. PMID- 15879712 TI - Non-pungent Capsicum contains a deletion in the capsaicinoid synthetase gene, which allows early detection of pungency with SCAR markers. AB - The capsaicinoid synthetase (CS) gene cosegregated perfectly with the C locus, which controls the presence of pungency, in 121 F2 individuals from a cross between 'ECW123R' and 'CM334', both of Capsicum annuum. We concluded that CS and C are tightly linked. Sequence analysis of the genes of four pungent and four non pungent pepper lines showed that the non-pungent peppers had a 2,529 bp-deletion in the 5' upstream region of CS. We have developed molecular markers of the C locus to detect pungency at the seedling stage. Based on the deleted sequence, we developed five SCAR markers, two of them being codominant. These SCAR markers will be useful for easy, accurate, and early detection of non-pungent individuals in breeding programs. PMID- 15879713 TI - Endoplasmic reticulum Ca2+ store: regulation of Ca2+ release and reuptake by intracellular and extracellular Ca2+ in pancreatic acinar cells. AB - We investigated the effect of cytosolic and extracellular Ca2+ on Ca2+ signals in pancreatic acinar cells by measuring Ca2+ concentration in the cytosol([Ca2+]c) and in the lumen of the ER([Ca2+]Lu). To control buffers and dye in the cytosol, a patch-clamp microelectrode was employed. Acetylcholine released Ca2+ mainly from the basolateral ER-rich part of the cell. The rate of Ca2+ release from the ER was highly sensitive to the buffering of [Ca2+]c whereas ER Ca2+ refilling was enhanced by supplying free Ca2+ to the cytosol with [Ca2+]c clamped at resting levels with a patch pipette containing 10 mM BAPTA and 2 mM Ca2+. Elevation of extracellular Ca2+ to 10 mM from 1 mM raised resting [Ca2+]c slightly and often generated [Ca2+]c oscillations in single or clustered cells. Although pancreatic acinar cells are reported to have extracellular Ca2+-sensing receptors linked to phospholipase C that mobilize Ca2+ from the ER, exposure of cells to 10 mM Ca2+ did not decrease [Ca2+]Lu but rather raised it. From these findings we conclude that 1) ER Ca2+ release is strictly regulated by feedback inhibition of [Ca2+]c, 2) ER Ca2+ refilling is determined by the rate of Ca2+ influx and occurs mainly in the tiny subplasmalemmal spaces, 3) extracellular Ca2+-induced [Ca2+]c oscillations appear to be triggered not by activation of extracellular Ca2+ sensing receptors but by the ER sensitised by elevated [Ca2+]c and [Ca2+]Lu. PMID- 15879714 TI - Methylation of CpG islands in the rat 7-dehydrocholesterol reductase promoter suppresses transcriptional activation. AB - In mammals, 7-dehydrocholesterol reductase (Dhcr7) is the terminal enzyme in cholesterol biosynthesis. We previously reported that the Dhcr7 proximal promoter (-179 to +1), which contains CpG islands, is responsible for sterol-mediated expression of the rat gene. In the present study, we examined whether methylation of this region affects the transcriptional activity of the Dhcr7 gene. In vitro DNA methylation of the Dhcr7 promoter and luciferase-reporter assays showed that DNA methylation of the CpG islands suppressed transcription. Furthermore, treatment of the methylated Dhcr7 promoter with the demethylating agent, 5-aza-2' deoxycytidine (5-Aza-CdR), reversed the suppression of promoter activity. These results indicate that methylation of the CpG islands is an important transcriptional regulatory mechanism in the Dhcr7 promoter. PMID- 15879715 TI - Phylogenetic relationships among six vetigastropod subgroups (Mollusca, Gastropoda) based on 18S rDNA sequences. AB - Complete 18S rDNA sequences were determined for 10 vetigastropods in order to investigate the phylogeny of Vetigastropoda, which is controversial. These sequences were analyzed together with published sequences for nine other vetigastropods and two nerites. With the two nerites as outgroups, the phylogeny was inferred by three analytical methods, neighbor-joining, maximum likelihood, and maximum parsimony. The 18S rDNA sequence data support the monophyly of four vetigastropod superfamilies, the Pleurotomarioidea, the Fissurelloidea, the Haliotoidea, and the Trochoidea. The present results yield the new branching order: (Pleurotomarioidea (Fissurelloidea ((Scissurelloidea, Lepetodriloidea) (Haliotoidea, Trochoidea)))) within the vetigastropod clade. PMID- 15879716 TI - Effects of Ser2 and Tyr6 mutants of BAF53 on cell growth and p53-dependent transcription. AB - BAF53 is an actin-related protein that shuttles between nucleus and cytoplasm. In the nucleus, it constitutes an integral component of many chromatin-modifying complexes such as the SWI/SNF, TIP60, TRRAP, and TIP48/49 complexes. BAF53 is essential for growth, but its function remains elusive. BAF53 homologues from yeast to humans have a conserved N-terminal motif, MS_(G/A)(G/A)_(V/L)YGG, which is unique to these proteins. Previously we showed that over-expression of an N terminal deletion mutant of BAF53 (BAF53_deltaN) reduced the viability of HEK293 and HeLa cells. When we replaced the serine 2 and tyrosine 6 of this N-terminal motif with alanine, over-expression of the alanine-replaced BAF53 strongly impaired the growth of HEK293 cells whereas replacement with aspartate/glutamate had no effect. The alanine-replaced BAF53 mutants also stimulated p53-dependent transcription, in which the SWI/SNF and TRRAP complexes are involved. Our results demonstrate that serine 2 and tyrosine 6 play important roles in BAF53 activity. PMID- 15879717 TI - Cloning and expression of a farnesyl diphosphate synthase in Centella asiatica (L.) Urban. AB - A cDNA encoding farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2.5.1.10) was isolated from Centella asiacita (L.) Urban, using degenerate primers based on two highly conserved domains. A full-length cDNA clone was subsequently isolated by rapid amplification of cDNA ends (RACE) PCR. The sequence of the CaFPS (C. asiatica farnesyl diphosphate synthase) cDNA contains an open reading frame of 1029 nucleotides encoding 343 amino acids with a molecular mass of 39.6 kDa. The deduced CaFPS amino acid sequence exhibits 84, 79, and 72%, identity to the FPSs of Artemisia annua, Arabidopsis thaliana, and Oryza sativa, respectively. Southern blot analysis suggested that the C. asiatica genome contains only one FPS gene. An artificially expressed soluble form of the CaFPS was identified by SDS-PAGE. It had high specific activity and produced farnesyl diphosphate as the major isoprenoid. PMID- 15879719 TI - Can artificial neural networks be beneficial in diagnosing gastro-oesophageal reflux disease? PMID- 15879720 TI - Infliximab in refractory coeliac disease. AB - Coeliac disease is generally well controlled with gluten-free diet but a small proportion of patients require corticosteroids or immunomodulatory agents. Response to anti-tumour necrosis factor (anti-TNF) agents raises interesting questions about both the pathogenesis of coeliac disease and the mechanism of action of anti-TNF agents. Refractory coeliac disease poses a therapeutic challenge to clinicians and carefully selected patients may benefit from anti-TNF therapy. PMID- 15879721 TI - Artificial neural networks are able to recognize gastro-oesophageal reflux disease patients solely on the basis of clinical data. AB - BACKGROUND: Artificial neural networks (ANN) are modelling mechanisms that are highly flexible and adaptive to solve the non-linearity inherent in the relationship between symptoms and underlying pathology. OBJECTIVES: To assess the efficacy of ANN in achieving a diagnosis of gastro-oesophageal reflux disease (GORD) using oesophagoscopy or pH-metry as a diagnostic gold standard and discriminant analysis as a statistical comparator technique in a group of patients with typical GORD symptoms and with or without GORD objective findings (e.g. a positive oesophagoscopy or a pathological oesophageal pH-metry). METHODS: The sample of 159 cases (88 men, 71 women) presenting with typical symptoms of GORD, were subdivided on the basis of endoscopy and pH-metry results into two groups: GORD patients with or without oesophagitis, group 1 (N=103), and pH and endoscopy-negative patients in whom both examinations were negative, group 2 (N=56). A total of 101 different independent variables were collected: demographic information, medical history, generic health state and lifestyle, intensity and frequency of typical and atypical symptoms based on the Italian version of the Gastroesophageal Reflux Questionnaire (Mayo Clinic). The diagnosis was used as a dependent variable. Different ANN models were assessed. RESULTS: Specific evolutionary algorithms selected 45 independent variables, concerning clinical and demographic features, as predictors of the diagnosis. The highest predictive performance was achieved by a 'back propagation' ANN, which was consistently 100% accurate in identifying the correct diagnosis compared with 78% obtained by traditional discriminant analysis. CONCLUSION: On the basis of this preliminary work, the use of ANN seems to be a promising approach for predicting diagnosis without the need for invasive diagnostic methods in patients suffering from GORD symptoms. PMID- 15879722 TI - Is cytokeratin immunoreactivity useful in the diagnosis of short-segment Barrett's oesophagus in Korea? AB - BACKGROUND: Cytokeratin 7/20 staining has been reported to be helpful in diagnosing Barrett's oesophagus and gastric intestinal metaplasia. However, this is still a matter of some controversy. OBJECTIVE: To determine the diagnostic usefulness of cytokeratin 7/20 immunostaining for short-segment Barrett's oesophagus in Korea. METHODS: In patients with Barrett's oesophagus, diagnosed endoscopically, at least two biopsy specimens were taken from just below the squamocolumnar junction. If goblet cells were found histologically with alcian blue staining, cytokeratin 7/20 immunohistochemical stains were performed. Intestinal metaplasia at the cardia was diagnosed whenever biopsy specimens taken from within 2 cm below the oesophagogastric junction revealed intestinal metaplasia. Barrett's cytokeratin 7/20 pattern was defined as cytokeratin 20 positivity in only the superficial gland, combined with cytokeratin 7 positivity in both the superficial and deep glands. RESULTS: Barrett's cytokeratin 7/20 pattern was observed in 28 out of 36 cases (77.8%) with short-segment Barrett's oesophagus, 11 out of 28 cases (39.3%) with intestinal metaplasia at the cardia, and nine out of 61 cases (14.8%) with gastric intestinal metaplasia. The sensitivity and specificity of Barrett's cytokeratin 7/20 pattern were 77.8 and 77.5%, respectively. CONCLUSION: Barrett's cytokeratin 7/20 pattern can be a useful marker for the diagnosis of short-segment Barrett's oesophagus, although the false positive or false negative rate is approximately 25%. PMID- 15879723 TI - Are lower gastrointestinal investigations necessary in patients with coeliac disease? AB - BACKGROUND: Colonoscopy may be indicated in patients with coeliac disease who present with iron deficiency anaemia or in coeliac disease patients who have persisting diarrhoea despite being on a gluten-free diet. However, there are limited data to support this approach. METHODS: We prospectively recruited patients who were found to have coeliac disease, having been referred with newly diagnosed iron deficiency anaemia. We also recruited a second group of patients with known coeliac disease. These patients had persisting diarrhoea despite being on a gluten-free diet for 6 months. All patients had colonoscopy and were matched with controls (without coeliac disease) who had similar indications for colonoscopy. RESULTS: Ninety-eight consecutive new patients with coeliac disease and concurrent iron deficiency anaemia had colonoscopy performed. Twelve (12.2%) had pathology, three of which were carcinomas. This diagnostic yield was not significantly different from the findings in the control group 62/362 (17.1%) P=0.24. In coeliac disease patients with persisting diarrhoea (n=37), the diagnostic yield at colonoscopy was 1/37 (2.7%). This was significantly lower than our findings in the control group with chronic diarrhoea 55/390 (14%) P=0.05. CONCLUSION: Colonoscopy should be considered in patients with coeliac disease (over the age of 45 years) who present with iron deficiency anaemia. Whilst, for coeliac disease patients with persisting diarrhoea (on a gluten-free diet) in the absence of sinister symptoms, a flexible sigmoidoscopy may be the initial investigation in order to exclude microscopic colitis. However, further larger prospective studies are required to evaluate this approach. PMID- 15879724 TI - Worse clinical course of disease in Crohn's patients with previous appendectomy. AB - OBJECTIVES: The aim of the present study was to assess the effect of previous appendectomy in a series of Crohn's disease (CD) patients on the clinical characteristics and course of disease. METHODS: Demographic and clinical data were retrospectively analysed for 129 consecutive outpatients (68 men and 61 women, median age 38 years) with CD. For each patient, information concerning appendectomy, indication for surgery (acute/chronic) and the date of surgery were recorded. The date of the appendectomy in relation to the date of CD diagnosis was carefully assessed in order to evaluate the precise relationship between the two events. A total of 129 CD patients who had not undergone previous appendectomy served as controls. The severity of disease was assessed retrospectively by evaluating the need for systemic steroids, immunosuppressants and surgical treatment for CD, particularly resective procedures. RESULTS: Forty one CD patients (31.8%) underwent appendectomy before the diagnosis of disease. Appendectomy before diagnosis showed a negative association with colonic disease localization and with articular manifestations. In addition, the 41 patients with previous appendectomy had a significantly greater risk of surgery, particularly resective. Multivariate analysis confirmed appendectomy performed before diagnosis as an independent risk factor for surgery; on the contrary, colonic site and inflammatory type of disease were independent factors protecting against surgery. Although current smokers were at an increased risk of surgical treatment, a smoking habit alone did not seem to be relevant at the multivariate analysis. CONCLUSION: The results of this study indicate a worse clinical course of CD in patients appendicectomized before diagnosis. PMID- 15879725 TI - Ten years experience with piezoelectric extracorporeal shockwave lithotripsy of gallbladder stones. AB - BACKGROUND: A critical review of the experience with extracorporeal shockwave lithotripsy (ESWL) of gallbladder stones is needed to clarify whether this method should continue to be applied to patients. METHODS: Patients with symptomatic gallbladder stones were treated by piezoelectric ESWL according to a prospective protocol between 1988 and 1997. ESWL treatment was limited to a maximum of three (solitary stones <20 mm diameter) to five sessions (larger solitary or multiple stones) and 3000 pulses per session. Univariate and multivariate analyses of pretreatment and treatment variables were performed to investigate their impact on fragmentation efficacy and stone clearance. A tree-based analysis was used to identify prognostically homogenous subgroups of individuals with maximum benefit from ESWL. RESULTS: Four hundred and eight patients, 76% female and 24% male, with a mean age of 46 (SD, 13) years, were selected for evaluation. Cox regression analysis identified three pretreatment variables with significant prognostic impact: (1) number of gallstones >1 (relative risk, 2.6 (95% CI, 1.9 3.5)), (2) size of stones >17 mm (1.7 (1.4-2.2)), and (3) computed tomography (CT) density of stones >55 Hounsfield units (H) (1.4 (1.1-1.8)). According to tree-based analysis, the stone clearance rate after 1 year was 85% (95% CI, 75 91%) for solitary stones <16 mm, 79% (70-86%) for solitary stones > or =16 mm with a CT density <84 H, 45% (32-55%) for solitary stones > or =16 mm with a CT density > or =84 H, and 42% (30-51%) for multiple stones. Five years after stone clearance, recurrence occurred in 43% of patients (95% CI, 39-47%). CONCLUSIONS: ESWL treatment showed an acceptable stone clearance in the case of small solitary gallbladder stones (<16 mm) or larger solitary stones with a CT density <84 H, but a very low success rate in the case of multiple stones. The poor long-term success, however, is an important argument against the use of ESWL of gallbladder stones. PMID- 15879726 TI - Changes in characteristics and outcome of acute upper gastrointestinal haemorrhage: a comparison of epidemiology and practices between 1996 and 2000 in a multicentre French study. AB - OBJECTIVE: To evaluate the main changes in characteristics, practices and outcome between 1996 and 2000 in patients admitted for an acute upper gastrointestinal haemorrhage (AUGIH). PATIENTS AND METHODS: All consecutive patients (n=1165) admitted for an AUGIH in four French administrative areas were entered into two separate 6-month studies conducted in 1996 (n=712) and 2000 (n=453). Epidemiological and biological characteristics, endoscopic haemostatic procedures and outcomes were compared. RESULTS: Patient characteristics remained unchanged between the two studies; the two main bleeding lesions were peptic ulcer and oesophagogastric varices (30.2 versus 31.1% and 22.5 versus 20.3%). The use of non-steroidal anti-inflammatory drugs or aspirin was more frequent in 2000 (26.5 versus 32.6%; P<0.03). Proton pump inhibitor preventative therapy was administered in less than 15% of patients with a high risk of peptic ulcer bleeding in each period. In patients admitted for varices bleeding, the use of endoscopic haemostatic ligation increased (17.1 versus 40%; P<0.001), with a concomitant decrease in endoscopic sclerotic therapy (76.1 versus 37.5%; P<0.001). We observed a significant decrease in AUGIH mortality in the whole group (11.7 versus 7.2%; P=0.03), and particularly in the subgroup of cirrhotic patients (19.5 versus 11.1%; P=0.05) whatever the source of their bleeding. CONCLUSION: Our time-trend evaluation of changes in AUGIH characteristics revealed that peptic ulcer and varices were still the two most frequent bleeding lesions. In patients with varices bleeding, endoscopic ligation became the routine standard treatment instead of varices sclerosis. The mortality rate decreased significantly over the 5-year study period in the whole group and particularly in the subgroup of cirrhotic patients. PMID- 15879727 TI - Long-term outcome of conservative treatment in patients with diverticulitis of the sigmoid colon. AB - INTRODUCTION: The indication for surgery after conservative treatment of acute diverticulitis is still under debate. This is partly as a result of limited data on the outcome of conservative management in the long run. We therefore aimed to determine the long-term results of conservative treatment for acute diverticulitis. METHODS: The records of all patients treated at our institution for diverticulitis between 1985 and 1991 were reviewed (n=363, median age 64 years, range 29-93). Patients who received conservative treatment were interviewed in 1996 and 2002 [follow-up time 7 years 2 months (range 58-127 months) and 13 years 4 months (range 130-196 months). RESULTS: A total of 252 patients (69%) were treated conservatively, whereas 111 (31%) were operated on. At the first follow-up, 85 patients treated conservatively had died, one of them from bleeding diverticula. A recurrence of symptoms was reported by 78 of the remaining 167 patients, and 13 underwent surgery. At the second follow-up, one patient had died from sepsis after perforation during another episode of diverticulitis. Thirty-one of the 85 patients interviewed reported symptoms and 12 had been operated on. In summary, at the second follow-up interview, 34% of patients treated initially had had a recurrence and 10% had undergone surgery. No predictive factors for the recurrence of symptoms or later surgery could be determined. CONCLUSION: Despite a high rate of recurrences after conservative treatment of acute diverticulitis, lethal complications are rare. Surgery should thus mainly be undertaken to achieve relief of symptoms rather than to prevent death from complications. PMID- 15879728 TI - Effects of lamivudine therapy on the glucose metabolism in children with chronic hepatitis B: first year follow-up results. AB - BACKGROUND AND AIM: Although the most common major toxicity of lamivudine has been pancreatitis, there is no report investigating possible impaired pancreatic functions, including glucose intolerance due to lamivudine therapy. The aim of this study was to evaluate the effects of lamivudine on the glucose metabolism in children. METHOD: Twenty-three children were included: eight patients were treated with lamivudine, others with both lamivudine and interferon-alpha. An oral glucose tolerance test (OGTT) was performed before the treatment, and after 6 and 12 months. RESULTS: After 6 and 12 months of the treatment four (18.4%) and eight (34.8%) patients had impaired OGTT, respectively. We did not find any relationship between impaired OGTT and age, gender, elevated amylase, abdominal pain and the mode of therapy (P>0.05). While mean glucose value after 2 h was higher than that of baseline, mean insulin concentrations and area under the curve values were not different (P<0.0001, P>0.05, and P>0.05, respectively). CONCLUSION: This is the first report demonstrating that lamivudine may impair the OGTT. Since at least 8.7% of our patients had persistently impaired OGTTs during the first year of the therapy, it may be reasonable to screen children before lamivudine therapy is started. PMID- 15879729 TI - Coombs-positive autoimmune hemolytic anemia in Crohn's disease. AB - BACKGROUND: Anemia often occurs in patients with inflammatory bowel diseases. However, hemolytic anemia is a rare complication and is associated with Coombs positive autoimmune disorders. There are several reports of autoimmune hemolytic anemia in patients with ulcerative colitis, whereas there are only four reports of this complication in patients with Crohn's disease. We report a case of a severe course Coombs-positive hemolytic anemia in a patient with Crohn's disease, which was refractory to medical treatment but resolved after subtotal colectomy. CASE REPORT: A 29-year-old patient was submitted to our clinic several times because of a severe course of inflammatory bowel disease and additionally a Coombs-positive autoimmune hemolytic anemia. Histology indicated severe Crohn's disease, but neither medical treatment with steroids, nor with methotrexate, cyclosporine or tumor necrosis factor-alpha antibody had been successful in resolving the intestinal inflammation and the hemolytic anemia. As colonoscopy revealed a pancolitis and dysplastic changes, even in the less inflamed areas of the colonic mucosa, subtotal colectomy was indicated. Half a year later we observed clinical and immunological signs of complete remission (no gastrointestinal symptoms, negative Coombs test). CONCLUSION: Autoimmune hemolytic anemia is a rare complication of inflammatory bowel disease and has been almost exclusively described in ulcerative colitis. The etiology is not yet completely understood. Presumably, the colon displays a role in the production of anti-erythrocyte antibodies. The therapy of choice in Crohn's associated hemolytic anemia is thought to be medical treatment with corticoid steroids. Some authors additionally prefer immunmodulators. However, in the case presented, colectomy (without splenectomy) was necessary to resolve refractory hemolysis and the severe course of Crohn's disease. PMID- 15879730 TI - Refractory coeliac disease: remission with infliximab and immunomodulators. AB - Coeliac disease is a T-cell-mediated enteropathy induced by gluten. A minority of patients who fail to respond to a gluten-free diet may require intervention with immunomodulating drugs. We report a case of refractory coeliac disease where remission was induced by the anti-tumour necrosis factor-alpha antibody infliximab and was maintained with prednisolone and azathioprine. PMID- 15879731 TI - Non-ruptured hydatid cyst can lead to death by spread of cyst content into bloodstream: an autopsy case. AB - Hydatid disease is a parasitic infestation caused by a tapeworm of the genus Echinococcus, and it is common in Mediterranean regions. Cystic lesions cause symptoms via compressing adjacent organs or may be totally silent. Morbidity is usually secondary to free rupture of the echinococcal cyst with or without anaphylaxis, infection of the cyst or dysfunction of affected organs. The cyst of Echinococcus granulosus is commonly located in the liver and frequently causes no symptoms. Anaphylactic reactions as a result of cyst perforation generally occur during interventions such as needle aspiration or open surgery; however, the spillage of cyst fluid with intravascular spread resulting from trauma may also trigger anaphylaxis, and rare case reports of this kind are present in the literature. We report the case of a 17-year-old man who was admitted to the public hospital with a sudden onset of nausea, vomiting and fainting. After a short period of intervention in the emergency department he died. As the cause of his sudden death was unknown, a forensic autopsy was carried out by the Forensic Council of Turkey. The autopsy revealed a macroscopically non-ruptured hydatid cyst in the liver and laryngeal oedema. In histopathological examination, two scolices in the pulmonary artery and inflammatory infiltration mainly composed of mast cells in the larynx were detected. Sudden death in this case was attributed to anaphylactic shock caused by intravascular spread of the cyst contents. PMID- 15879732 TI - Acute pancreatitis induced by traditional acupuncture therapy. AB - Although rare, it is possible for acute pancreatitis to develop after blunt or penetrating abdominal trauma. We report here a very rare case of acute pancreatitis induced by traditional acupuncture therapy. A 42-year-old woman with a low body mass index had suffered from functional dyspepsia for one year. She visited an acupuncture clinic and underwent long needle (13 cm) and gold thread needle (3 mm) acupuncture therapy. Five hours later, she presented at our emergency room complaining of severe periumbilical pain. Levels of serum amylase (1162 U/l, normal <220 U/l), and lipase (5195 IU/l, normal <60 IU/l) were high, and an abdominal computed tomography scan showed a diffusely swollen pancreas and ill-defined infiltration of the peripancreatic fat, indications of possible acute pancreatitis. Multiple small acupunctured gold thread needles were also found on the anterior abdominal wall and back muscles. After excluding other possible causes of pancreatitis, traumatic pancreatitis was diagnosed as an adverse effect of the long acupuncture needle therapy. PMID- 15879733 TI - Menetrier's disease presenting as an acute protein-losing gastroenteropathy in a 27-year-old man with Gaucher disease. AB - OBJECTIVES: To describe a unique case of a young man with Gaucher disease who was diagnosed with Menetrier's disease. BACKGROUND: After an acute episode of severe gastritis, the patient developed hypoalbuminemia and protein-losing gastroenteropathy, and became unwell. STUDY: Endoscopy revealed an abnormal stomach, with rigid, thickened folds covered with viscous greyish exudates. Superficial biopsies revealed foveolar hyperplasia, acute and severe gastritis with massive inflammatory infiltrate of neutrophils in the lamina propria with pit abscess formation. Tissue cultures for Helicobacter pylori were negative. RESULTS: Snare deep particle biopsy revealed the typical features of Menetrier's disease. Enzyme replacement therapy for Gaucher disease was started. CONCLUSION: This case poses a dilemma because the patient improved spontaneously, and as such is dissimilar to other adults who develop Menetrier's disease because of an infection; it is hoped that he may also not be at risk of the potential malignancies that are correlated with adult Menetrier's disease. The value of enzyme treatment is considered. PMID- 15879734 TI - Carcinosarcoma of the biliary tract: two case reports and a review of the literature. AB - Carcinosarcoma of the biliary tract is extremely rare. Little is known about the natural course of these tumours, or the best available treatment. We present two cases of carcinosarcoma of the biliary tract, one of the gall bladder and one of the common bile duct, followed by a review of the literature. PMID- 15879735 TI - Irritable bowel syndrome and endometriosis. PMID- 15879740 TI - Altering metabolism. AB - A significant proportion of the mortality and morbidity of severe burns is attributable to the ensuing hypermetabolic response. This response can last for as long as 1 year after injury and is associated with impaired wound healing, increased infection risks, erosion of lean body mass, hampered rehabilitation, and delayed reintegration of burn survivors into society. Pharmacologic and nonpharmacologic strategies may be used to reverse the catabolic effect of thermal injury. Nonpharmacologic strategies include early excision and wound closure of burn wound, aggressive treatment of sepsis, elevation of the environmental temperature to thermal neutrality (31.5 +/- 0.7 degrees C), high carbohydrate, high protein continuous enteral feeding, and early institution of resistive exercise programs. Pharmacologic modulators of the postburn hypermetabolic response may be achieved through the administration of recombinant human growth hormone, low-dose insulin infusion, use of the synthetic testosterone analog, oxandrolone, and beta blockade with propranolol. This review article discusses these modulators of postburn metabolism. PMID- 15879741 TI - Respiratory care of the burn patient. PMID- 15879742 TI - The burn edema process: current concepts. AB - Massive tissue edema after thermal injury is a well-recognized entity. Although this process is responsible for the patient's large fluid needs during resuscitation and also for local problems, such as a compartment syndrome, there have been no effective treatment modalities introduced into clinical care to control the degree of edema. A review of what is now known about the edema process is presented here, including attempted prevention and treatment modalities. The pathogenesis involves changes in most of the physical forces controlling fluid flux across the capillary and also how fluid accumulates in the interstitium. Increased capillary permeability to protein is but one of these changes. The presence of an initial profound negative interstitial pressure "sucking" fluid into the tissues and a marked increase in interstitial space compliance are equally important components. A host of mediators, especially oxidants, have been reported to cause these physical changes, and some mediator inhibitors appear to be of benefit, especially antioxidants. However, few clinical trials, aimed at decreasing edema, have been performed. With these new insights into the edema process, future prevention and treatment modalities can be developed. PMID- 15879743 TI - Methamphetamine laboratory explosions: a new and emerging burn injury. AB - The proliferation of clandestine methamphetamine laboratories (meth labs) as a result of the growing popularity of the drug has resulted in an increasing incidence of burn injuries associated with laboratory accidents. We undertook this study to characterize these injuries. Fifteen consecutive patients were identified and case-matched by age and TBSA to 45 control subjects. Most meth lab patients were men, Caucasian, unemployed, and positive for polysubstance abuse. Resuscitation requirements were 1.8 times greater in these patients. There was a higher incidence of inhalational injury corresponding to higher intubation and tracheostomy rate and longer ventilator days among meth lab patients. The rate of nosocomial pneumonia, skin graft loss, and mortality were not different between the two groups. Meth lab injury is unique and requires more critical care resources. It also is associated with lack of insurance coverage and poor follow up after injury. This injury has a significant impact not only on patients but also on the healthcare system. PMID- 15879744 TI - Pattern of dysphagia recovery after thermal burn injury. AB - Providing nutritional support after thermal burn injury is a critical element in achieving successful patient outcomes. The medical records of 28 patients admitted to an acute care burn unit and referred to speech pathology for a swallowing evaluation were reviewed for patterns of dysphagia care. Results revealed a strong positive linear relationship between percent body burns (r = .71), number of days with a tracheostomy (r = .85), number of days on the ventilator (r = .94) and days to oral feeding. These data can be used for comparison with other models of care, such as early identification and intervention for dysphagia by the speech pathologist during the acute phase of recovery. PMID- 15879745 TI - Evaluation of cytotoxicity and antimicrobial activity of Acticoat Burn Dressing for management of microbial contamination in cultured skin substitutes grafted to athymic mice. AB - Cultured skin substitutes (CSS) have become a useful adjunctive treatment for closure of burn wounds, but CSS are avascular and remain susceptible to microbial destruction longer than split-thickness skin grafts. Irrigation of CSS grafted to burn wounds with a topical antimicrobial solution (TAS) has been shown to promote engraftment of CSS, but TAS usage has potential limitations. Acticoat Burn Dressing (Acticoat; Westaim Biomedical, Exeter, NH) is a silver-coated barrier dressing reported to exhibit antimicrobial activity and to reduce infection in partial-thickness and full-thickness wounds. This study evaluated the cytotoxicity of Acticoat with CSS and the efficacy of Acticoat for the management of microbial contamination in CSS grafted to full-thickness wounds in athymic mice. The cytotoxicity of Acticoat was assessed in preliminary studies after 1 week of exposure to CSS during in vitro maturation or healing on wounds in athymic mice. Histologies were analyzed and cellular viability in the CSS was determined by MTT conversion on days 0, 1, and 7 of Acticoat exposure. At 1, 2, 3, and 4 weeks after grafting, wounds were traced, and areas of healing CSS were calculated by image analysis. At 4 weeks, wound biopsies were evaluated and scored for engraftment of human cells. In a subsequent study, wounds were inoculated with strain SBI-N of Pseudomonas aeruginosa at 1 x 10(5) cfu/wound before the application of CSS or inoculated onto the surface of Acticoat. At 4 weeks, swab cultures were collected from the surface of CSS and scored for the presence of SBI-N. Statistical significance was accepted at the 95% confidence level (P <.05). The data show that exposure in vitro of CSS to Acticoat was cytotoxic within 1 day, but 1 week of exposure in vivo did not injure CSS or inhibit wound healing. Contaminated wounds treated with Acticoat healed similarly to control treatments, with comparable rates of engraftment, and detection of SBI N on the surface of only one graft. No SBI-N was detected on CSS after inoculation onto the surface of Acticoat. These results suggest that Acticoat may be suitable as a protective dressing to reduce environmental contamination of CSS, if used in conjunction with additional antimicrobials to control organisms present in the wound. PMID- 15879746 TI - Efficacy of moist exposed burn ointment on burns. AB - In this study, we sought to test the medical efficacy of a Chinese medical herb product, moist exposed burn ointment (MEBO), on wound healing rate and infection control in burn injury. Standardized deep burn wounds were created on the back skin of rats by applying a hot brass bar for 12 to 18 seconds. MEBO was applied four times per day and compared with petroleum jelly, silver sulfadiazine, and dry exposure therapy. Under such a controlled setting, although MEBO had a better wound healing rate than the dry exposure treatment, it did not show the medical advantage statistically, as has been claimed, over the other two treatments (P > .05), either in terms of wound healing rate or bacterial control. We conclude that the MEBO is not suitable for deep burn wound treatment, particularly when infection is a concern. PMID- 15879747 TI - Cardiac myocyte accumulation of calcium in burn injury: cause or effect of myocardial contractile dysfunction. AB - Myocardial calcium accumulation and myocardial injury occur after burn trauma. However, whether altered calcium dyshomeostasis occurs as a result of myocardial injury/dysfunction or whether altered calcium handling initiates myocardial injury and contractile abnormalities remains unclear. In addition, the specific mechanisms by which burn injury promotes calcium entry into cardiac myocytes, specifically L-type channels and the sodium-calcium exchanger, remain unclear. This study addressed the hypothesis that burn trauma promotes cardiomyocyte calcium accumulation, in part, via reverse mode function of the sodium/calcium exchanger and via L-type channels. Myocardial calcium accumulation, in turn, alters performance. Burn trauma (40% TBSA and sham burn for controls) was accomplished in Sprague-Dawley rats. Burns received fluid resuscitation (lactated Ringer's at 4 ml/kg/% burn). Hearts were harvested at several time points after burn injury (2, 4, 8, 12, 24, 48, 72 hours, and 8 days after burn) and were perfused with collagenase/bovine serum albumin-containing buffer to produce enzymatic digestion. Myocytes were then resuspended in MEM buffer, loaded with 2 mug/ml Fura 2AM for 45 minutes or 2 microg of sodium-binding benzofurzan isophthalate for 2 hours at room temperature in the dark. Cells were washed to remove extracellular dye and placed on a glass slide on the stage of a Nikon inverted microscope interfaced with Grooney optics. A computer-controlled filter changer allowed alternation between 340/380 excitation wavelengths; fluorescence was measured at 510 nm. Cardiac function (Langendorff) was measured in parallel groups at each time period (n = 6-7 hearts/time point). Cardiomyocyte accumulation of sodium occurred before alterations in myocyte calcium levels, and sodium/calcium dyshomeostasis preceded cardiac contraction deficits. Interventions that altered calcium flux through L-type channels (amlodipine) or sodium/calcium exchange (amiloride) attenuated burn-related myocyte calcium accumulation and improved contractile function. Our finding that myocyte sodium loading precedes myocyte calcium accumulation suggests a role for the reverse mode function of the sodium/calcium exchanger in burn trauma. PMID- 15879748 TI - Reproducibility of transpulmonary thermodilution measurements in patients with burn shock and hypothermia. AB - Thermal noise, including hypothermia and short-term variations in body temperature, has been reported to influence the accuracy and reproducibility of thermodilution measurements. This variation might theoretically limit the usefulness of this technique in patients with acute burns. We therefore sought to determine the reproducibility of hemodynamic parameters derived from arterial thermal dilution in patients with acute burns and hypothermia. This was a prospective, clinical study of 50 mechanically ventilated patients with burns involving more than 25% of the body surface area that were treated at an eight bed intensive care burn unit in a university-affiliated hospital. A total of 750 arterial thermodilution measurements were analyzed using the COLD system. Triplicate measurements of the intrathoracic blood volume, cardiac output, total blood volume, and extravascular lung water were performed at regular intervals during the first 48 hours after the thermal injury. Reproducibility was assessed by the coefficient of variation of the triplicate measurements. The correlation of variation was less than 10% at all measurement times for cardiac output, intrathoracic blood volume, and total blood volume. For the extravascular lung water, the coefficient of variation ranged from 9.5% to 12.9%. A maximum of 12.9% was found at 48 hours after burn. No correlation was found between body core temperature and the reproducibility of intrathoracic blood volume index (r = 0.145), cardiac index (r = 0.217), or extravascular lung water index (r = 0.167). The parameters derived from arterial thermodilution show a clinically sufficient reproducibility in patients with acute burns associated with thermal instability. PMID- 15879749 TI - Burn injuries in patients with dementia. PMID- 15879750 TI - Burn injury in patients with dementia: an impetus for prevention. AB - Current literature has reported an increase in the rates of morbidity and mortality in elderly dementia patients who have suffered from illnesses such as pneumonia or traumatic injuries such as falls, motor vehicle collisions, and other insults. The role of dementia in elderly burn patients has not been studied in depth. To assess the extent of this problem, a retrospective, case-control study of patients with dementia who were admitted to a large urban burn center was performed. The demographics, circumstance and severity of injury, critical care use, and discharge disposition of those patients admitted with dementia were reviewed and compared with the findings of age/burn size-matched controls. The results support the premise that burn injuries in this patient population can be severe. Although not statistically significant, 22.2% of the study group patients required ventilatory support, and 75% required monitoring in the intensive care unit compared with the 15.3% and 61.6% of control patients who required ventilatory support and monitoring in the intensive care unit, respectively. Also, although not statistically significant, the mortality rate of the study group was 25%, almost double that of the control group (13.8%). No other significant differences were observed. These findings support the need for assistance and supervision with daily activity and burn prevention education for this population. As our population ages and we are faced with caring for those with dementia, further burn prevention is warranted. PMID- 15879751 TI - Documenting changes in burn scars over time. PMID- 15879752 TI - Reliability testing of a new scar assessment tool, Matching Assessment of Scars and Photographs (MAPS). AB - A new scar-assessment tool, the Matching Assessment of Scars and Photographs (MAPS), which uses a set of reference photographs, a numeric scale, and location technique, was tested for its reliability in two stages. First, using five adults, three raters assessed 32 burns scars twice within 3 days. Subsequently, reliability was tested during a 6-month time frame, emphasizing the process of localizing test areas as scars changed and raters forgot the previous assessment. Three raters, from a pool of five, each made three assessments on 29 scars in seven subjects, on average 8 weeks apart. Inter-rater reliability was tested, using Kendall's Tau C and intraclass correlations, respectively, for stages 1 and 2. Agreement was good for border height (0.63-0.70 and 0.78), moderate to good for thickness (0.60-0.74 and 0.81), and good for color (0.55-0.71 and 0.79), whereas for surface it was fair (0.25-0.38 and 0.40). The localization technique was reliable (accuracy within 3 mm) in 93% to 96% of recordings. The MAPS tool is considered ready for clinical use. PMID- 15879753 TI - A case report of phaeohyphomycosis caused by Cladophialophora bantiana treated in a burn unit. AB - Black molds are a heterogeneous group of fungi that are distributed widely in the environment and that occasionally cause human infection. The spectrum of disease includes mycetomas, chromoblastomycosis, sinusitis, and superficial, cutaneous, subcutaneous, and systemic phaeohyphomycosis. Cladophialophora bantiana, an agent of phaeohyphomycosis, causes rare infections mainly of the central nervous system. Extracerebral involvement is uncommon, and only a few cases have been reported. We present the case of a 32-year-old immunosuppressed female who developed a cutaneous phaeohyphomycosis from C. bantiana. The patient was treated in a burn unit with wound care, surgical excision, grafting, and itraconazole. Patients with complex fungal infections represent yet another population with specialized needs that are adequately met in a verified burn center. PMID- 15879754 TI - Mortal hot spring burns: report of two cases. AB - This article describes two children who received deep partial and full-thickness mortal immersion scald burns while visiting a hot spring. The prevention of such accidents is most important, including the education of children and family members on the potential hazards inherent at these resorts. In addition, elevated retaining walls with closed upper surfaces of the pools should be reconstructed to avoid children slipping accidentally into hot springs. PMID- 15879755 TI - Hydrofluoric acid burns of head and neck. PMID- 15879759 TI - Subspecialty certification in pediatric urology. PMID- 15879760 TI - Tumor markers for renal cell carcinoma. PMID- 15879761 TI - Eliminating the current Bell curve to improve the results of radical prostatectomy. PMID- 15879762 TI - Bladder rehabilitation in children with spina bifida: state-of-the-"ART". PMID- 15879763 TI - Benign prostatic hyperplasia: patient perceptions and financial reality regarding the aging American prostate. PMID- 15879764 TI - Renal cell carcinoma 2005: new frontiers in staging, prognostication and targeted molecular therapy. AB - PURPOSE: Renal cell carcinoma (RCC) has traditionally been staged using a purely anatomical staging system. Although current staging systems provide good prognostic information, data published in the last few years has led to significant controversies as to whether further revisions are needed and whether improvements can be made with the introduction of new, more accurate and predictive prognostic factors not currently included in traditional staging systems. This review highlights such controversies and provides an update on current staging modalities, prognostic factors and targeted molecular therapy for RCC. MATERIALS AND METHODS: A comprehensive review of the peer reviewed literature was performed on the topic of current staging modalities, validated prognostic factors, predictive nomograms, molecular markers and targeted molecular therapy for RCC. RESULTS: A staging system for malignant disease such as RCC uses various characteristics of tumors to stratify patients into clinically meaningful categories, which can be used to provide patients with counseling regarding prognosis, select treatment modalities and determine eligibility for clinical trials. The TNM staging system is currently the most extensively used one. However, it has undergone recent systematic revision due to rapidly emerging data from longer patient followup. The identification of various histological and symptomatic factors has led groups at many centers to develop more comprehensive staging systems that integrate these factors and include patients with metastatic and local disease. While integrated staging systems have improved RCC staging, the recent discovery of molecular tumor markers is expected to revolutionize RCC staging in the future and lead to the development of new therapies based on molecular targeting. CONCLUSIONS: Staging systems for RCC serve as a valuable prognostic tool. Several new patient and tumor characteristics have been reported to be important prognostic factors and they have been integrated into current staging systems. In addition, the field of RCC is rapidly undergoing a revolution led by molecular markers and targeted therapies. With this information urologists will be updated with the most current and comprehensive staging strategies, and be provided with a glimpse of the molecular and patient specific staging and treatment paradigms that will in our opinion transform the future management of this malignancy. PMID- 15879765 TI - Contemporary appraisal of radical perineal prostatectomy. AB - PURPOSE: In the age of minimally invasive surgery there has been renewed interest in the perineal approach for the surgical treatment of prostate cancer. We reviewed recent publications regarding radical perineal prostatectomy (RPP) in an effort to define its role in the current management of localized prostate malignancy. At the same time we reviewed the relevant perineal anatomy and surgical approach necessary to perform this operation. MATERIALS AND METHODS: We performed a review of the literature with respect to RPP and included our own extensive experience with this operation, emphasizing patient selection, the current role of pelvic lymph node dissection, surgical anatomy, oncological outcomes and complications. RESULTS: RPP is an effective treatment for localized adenocarcinoma of the prostate with oncological outcomes similar to those of the retropubic technique. In comparison to RRP, patients undergoing RPP have less postoperative discomfort, more rapid return of bowel function, more rapid return to work and a decreased transfusion rate. In addition, RRP is now often performed with cavernous nerve sparing. Prostate specific antigen screening has made the rate of lymph node metastasis low enough to omit lymphadenectomy in many cases. CONCLUSIONS: There is still a role for RPP in the treatment of localized prostate cancer. Erectile dysfunction after nerve sparing and incontinence rates are similar to those of RRP. In addition, it is less morbid then RRP without being as technically challenging as laparoscopic radical prostatectomy. PMID- 15879766 TI - Defining recurrence after radiation for prostate cancer. AB - PURPOSE: We reviewed prostate specific antigen (PSA) definitions of recurrence after external beam radiation for prostatic cancer and related them to the definitions used for other treatment modalities. MATERIALS AND METHODS: The literature on defining recurrence after external beam radiation, brachytherapy and prostatectomy for prostate cancer was reviewed through a MEDLINE search to ensure completeness and the inclusion of all pertinent information. RESULTS: Although the definition, which is the current standard for estimating recurrence after external beam radiation, has proved to be a reasonable measure, alternative options that are more sensitive and specific have now been defined. Similar statistical testing and comparison must also be done for other treatment modalities since the choice of failure definitions has not been evaluated nearly as thoroughly for these therapies. As much as possible, outcome reporting should be done according to the same method to ensure fairness when comparisons are made. However, inherent differences between treatment modalities and their effect on PSA production must also be considered. CONCLUSIONS: With the latest available information from patients with long-term followup PSA definitions of tumor recurrence after external beam radiation for prostatic cancer must again be reviewed in a consensus conference format. The application of a universal PSA definition of tumor recurrence across multiple treatment modalities should also be explored. PMID- 15879767 TI - Urogenital surgery of the 15th century in Anatolia. AB - PURPOSE: We examined the urological procedures of the 15th century covered in the surgery textbook, Cerrahiyyetu'l Haniyye. MATERIALS AND METHODS: Three copies of the surgery textbook, Cerrahiyyetu'l Haniyye (Sultan's Surgery), by Serefeddin Sabuncuoglu, who lived in Turkey between 1385 and 1470, were evaluated. The colorfully miniaturized and illustrated textbook, written 536 years ago, covers a number of surgical disciplines, including urology. We evaluated urological sections with regard to the type of procedures, definitions and approaches for certain diseases and surgical tools used for urological operations. RESULTS: The textbook reviews the surgical treatment of urological conditions such as varicocele, hydrocele, hermaphroditism, imperforated urinary meatus, meatal stenosis, hypospadias, penile and scrotal lesions, and circumcision techniques. It includes definitions of diseases, etiologies and surgical therapies, and describes surgical instruments. The author also illustrated surgical approaches and instruments. CONCLUSIONS: A treasure trove of surgical knowledge, Cerrahiyyetu'l Haniyye has enlightened urologists from the 15th century to the present day. PMID- 15879768 TI - Renal medullary carcinoma and ABL gene amplification. AB - PURPOSE: We characterized the clinical course of renal medullary carcinoma (RMC) and performed an expanded analysis of BCR-ABL. MATERIALS AND METHODS: The literature was searched for all reports of RMC. New cases at Northwestern University are described and relevant clinical information was abstracted. BCR and ABL genes, and ABL protein were evaluated by fluorescence in situ hybridization and immunohistochemical analysis, respectively. RESULTS: A total of 95 cases were identified. Mean age at diagnosis was 19 years, the male-to-female ratio was 1.9:1.0, 90% of all patients were black, 98% had an abnormality in a least 1 hemoglobin gene (ie sickle cell trait, SC disease or sickle cell disease) and mean survival was 19 weeks. Two patients (3%) without metastasis were long term survivors. The response to chemotherapy was poor. One patient treated with thalidomide survived for 52 weeks. The ABL gene was amplified a mean +/- SEM of 1.9 +/- 0.1-fold in all 3 cases evaluated, while ABL protein was increased in 2 of 3 evaluated. No evidence of BCR-ABL translocation was detected. CONCLUSIONS: RMC is typically seen in young individuals with the sickle cell trait. It is diagnosed when metastatic, is not responsive to systemic therapy and rapidly causes death. Because cure appears possible with early diagnosis, increased awareness of the disease could make an impact. The use of thalidomide or newer anti-angiogenesis agents should be considered for advanced disease. The role of ABL amplification with respect to etiology and as a therapeutic target should be investigated further. PMID- 15879769 TI - Independent validation of the 2002 American Joint Committee on cancer primary tumor classification for renal cell carcinoma using a large, single institution cohort. AB - PURPOSE: The primary tumor classification for renal cell carcinoma (RCC) was updated by the American Joint Committee on Cancer in 2002. To date the new classification has not been validated using an independent group of patients and, therefore, its accuracy for predicting patient outcome is unknown. In the current study we evaluated the 2002 primary tumor classification and compared its predictive ability with that of the 1997 classification. MATERIALS AND METHODS: We studied 2,746 patients treated with radical nephrectomy or nephron sparing surgery for unilateral, sporadic RCC between 1970 and 2000. Cancer specific survival was estimated using the Kaplan-Meier method. The predictive abilities of the 1997 and 2002 classifications were compared using the concordance index. RESULTS: There were 812 deaths from RCC a mean of 3.3 years following nephrectomy. Median followup in patients still alive at last followup was 9 years. Estimated 5-year cancer specific survival rates by the 2002 tumor classification were 97%, 87%, 71%, 53%, 44%, 37% and 20% in patients with pT1a, pT1b, pT2, pT3a, pT3b, pT3c and pT4 RCC, respectively. The concordance index for the association between the 2002 classification and death from RCC was 0.752 compared with 0.737 for the 1997 classification, indicating that the 2002 version contained more predictive ability. CONCLUSIONS: Our data suggest that the 2002 primary tumor classification with pT1 cancers subclassified into pT1a and pT1b provides excellent stratification of patients according to cancer specific survival and it has a predictive ability that is superior to that of the 1997 classification. PMID- 15879770 TI - Comparison of presentation and outcome for patients 18 to 40 and 60 to 70 years old with solid renal masses. AB - PURPOSE: We compared histological subtype, pathological features and outcome of patients with solid renal masses who were 18 to 40 years old vs patients who were 60 to 70 years old. MATERIALS AND METHODS: We conducted a retrospective review of the Mayo Clinic Nephrectomy Registry from 1970 to 2000, and identified 124 patients 18 to 40 years old and 1,067 patients 60 to 70 years old available for analysis. RESULTS: There was no significant difference in the incidence of benign solid renal masses between patients 18 to 40 years old and those 60 to 70 years old (13.7% vs 10.2%). Among patients with renal cell carcinoma (RCC), younger patients were more likely to have chromophobe RCC (13.1% vs 3.6%) and less likely to have clear cell RCC (70.1% vs 81.5%) than older patients. Among patients with clear cell RCC, younger patients were more likely to have stage pT2b or lower tumors (82.7% vs 69.9%) and a higher incidence of cystic clear cell RCC (10.7% vs 2.2%) than older patients. Younger patients had an improved cancer specific survival compared with older patients but this difference was not statistically significant (risk ratio 0.71, p =0.127). CONCLUSIONS: We found that patients 18 to 40 years old were more likely to have chromophobe and less likely to have clear cell RCC compared with patients 60 to 70 years old. We did not identify a higher incidence of papillary RCC in younger patients. Patients with clear cell RCC 18 to 40 years old had a higher incidence of low stage and cystic tumors compared with patients 60 to 70 years old, features which have been shown to have a favorable prognosis. These factors likely contributed to improved cancer specific survival for younger patients. PMID- 15879771 TI - Renal cell carcinoma with renal vein and inferior vena caval involvement: clinicopathological features, surgical techniques and outcomes. AB - PURPOSE: We present a contemporary review of patients with renal cell carcinoma (RCC) in whom renal vein/inferior vena caval thrombus was treated with radical nephrectomy and thrombectomy. MATERIALS AND METHODS: A total of 220 patients underwent radical nephrectomy for RCC at our institution from 1998 to 2002. Of them 49 patients with renal vein/inferior vena caval involvement (T3b/c) were selected for review. We evaluated demographics, presenting symptoms, imaging modalities, clinical staging, pathological features, adjuvant treatment and clinical outcomes. We also evaluated surgical incisions, liver mobilization procedures, blood loss, transfusion requirements and perioperative mortality/morbidity. RESULTS: Gross hematuria was the most common presenting symptom, seen in 22 patients (45%), followed by constitutional symptoms in 8 (16%). Stage T3b/c was clinically diagnosed in 44 patients, while 2 had T2 and 2 had T4 disease. A subcostal incision was made in 30 patients, a chevron incision was made in 18, and a sternotomy and flank incision were made in 1. Liver mobilization was necessary in 13 patients and 2 required a Pringle maneuver. Cardiopulmonary bypass was performed in a single patient. Lymph node involvement was seen in 4 patients (8%) and distant metastases were present in 10 (20%). Median tumor size was 10 cm. Clear cell carcinoma was most common, as seen in 42 patients. Early (30-day) mortality in this series was 8%. At a median followup of 15 months 21 patients (43%) were without evidence of disease, 14 (29%) had disease, 8 (16%) had died of disease and 2 (4%) had died of other causes. None of the patients with lymph node involvement survived beyond 8 months after surgery. Tumor grade and T stage were found to be significant negative predictors of survival on univariate analysis. CONCLUSION: Most patients with RCC and tumor thrombus are symptomatic at presentation and metastatic disease at presentation is not uncommon. These results support the role of aggressive surgical treatment as the best initial management of these tumors. The majority of tumors can be approached and safely controlled without the need for a thoracoabdominal incision. While surgery provides modest disease-free survival, most patients should be offered immunotherapy, particularly those with advanced stage, grade, nodal involvement or metastases. PMID- 15879772 TI - Renal cryoablation: outcome at 3 years. AB - PURPOSE: We report intermediate term oncological followup data on 56 patients undergoing laparoscopic renal cryoablation, of whom each completed a 3-year followup. MATERIALS AND METHODS: Since September 1997, 56 patients undergoing laparoscopic renal cryoablation have completed a followup of 3 years each. The postoperative followup protocol comprised serial magnetic resonance imaging (MRI) at 1 day, months 1, 3, 6, 12, 18 and 24, and yearly thereafter for 5 years. Computerized tomography guided needle biopsy of the cryolesion was performed 6 months postoperatively and repeated if MRI findings were abnormal. Followup data were obtained prospectively. RESULTS: For a mean renal tumor size of 2.3 cm mean intraoperative size of the created cryolesion was 3.6 cm. Sequential mean cryolesion size on MRI on postoperative 1 day, and at 3 and 6 months, and 1, 2 and 3 years was 3.7, 2.8, 2.3, 1.7, 1.2 and 0.9 cm, representing a 26%, 39%, 56%, 69% and 75% percent reduction in cryolesion size at 3 and 6 months, and 1, 2 and 3 years, respectively. At 3 years 17 cryolesions (38%) had completely disappeared on MRI. Postoperative needle biopsy identified locally persistent/recurrent renal tumor in 2 patients. In the 51 patients undergoing cryotherapy for a unilateral, sporadic renal tumor 3-year cancer specific survival was 98%. There was no open conversion, kidney loss, urinary fistula, dialysis requirement, or perirenal or port site recurrence in any patients. CONCLUSIONS: Three-year outcomes following renal cryoablation are encouraging. Longer term (5-year) data are necessary to determine the proper place of renal cryotherapy among minimally invasive, nephron sparing options. PMID- 15879773 TI - Does the management of bladder perforation during transurethral resection of superficial bladder tumors predispose to extravesical tumor recurrence? AB - PURPOSE: We evaluated the impact of a bladder perforation during transurethral resection of superficial bladder tumor on extravesical tumor recurrence and patient prognosis. We also defined potential risk factors for extravesical recurrence prospectively giving emphasis to the management of the perforation. MATERIALS AND METHODS: The medical records of 3,410 patients were reviewed. Parameters recorded included patient age and sex, tumor stage, grade, number, size and location at the time of perforation, the type of bladder perforation (extraperitoneal vs intraperitoneal) and the way the perforation was managed (open surgical repair vs conservative treatment). Logistic regression analysis was used to identify risk factors for extravesical recurrence. Cox regression analysis was used to compare cancer specific survival. RESULTS: A total of 34 cases of bladder perforation were recorded, 4 patients were treated with open surgery and 30 treated conservatively. The 4 patients who underwent open surgery presented with extravesical recurrence after a mean followup of 7.5 months. The remaining 30 patients had no evidence of extravesical recurrence after a mean followup of 60 months (p <0.001). Of the patients with extravesical relapse 3 died of disease. The surgical management of bladder perforation was the best predictor of extravesical recurrence (p <0.001, r = 1.13), followed by an intraperitoneal localization of the perforation (p =0.0003, r = 0.67) and tumor size (p =0.01, r = 0.42). CONCLUSIONS: Surgical repair of a bladder perforation during transurethral resection of bladder tumor increases the risk of extravesical tumor cell recurrence and negatively affects patient prognosis. PMID- 15879774 TI - Laparoscopic radical cystectomy in the female. AB - PURPOSE: We detail the technique of completely intracorporeal laparoscopic radical cystectomy in the female patient, which has previously not been well described in the literature. Additionally, perioperative and short-term oncological outcome data are presented. MATERIALS AND METHODS: Since 2000, 11 female patients underwent laparoscopic radical cystectomy for bladder carcinoma. Data were collected with respect to patient demographics, perioperative outcomes and short-term oncological followup. RESULTS: Mean patient age was 66 years (range 42 to 80) and the mean body mass index was 25 (range 17 to 34). Mean total operative time was 8.5 hours (range 6.5 to 10.5), including an average radical cystectomy time of 2 to 2.5 hours. Estimated blood loss was 489 cc (range 150 to 1,000). Reconstructive procedures were an ileal conduit in 8 patients, Studer orthotopic neobladder in 2 and continent Indiana pouch in 1. Mean hospital stay was 6 days (range 5 to 12). No case was converted to open surgery. Complications occurred in 4 patients, including internal hernia requiring laparotomy 19 days postoperatively, deep vein thrombosis with pulmonary embolism, dehydration and urinary tract infection in 1 each. At a median followup of 7.1 months (range 1 to 19) 8 patients (73%) had no evidence of disease, 1 (9%) had metastasis and 2 (11%) had died. CONCLUSIONS: Laparoscopic radical cystectomy in the female is technically efficacious. Our techniques of anterior pelvic exenteration, and uterus/fallopian tube/ovary and nerve sparing laparoscopic female radical cystectomy are presented. Short-term functional and oncological outcomes appear favorable. PMID- 15879775 TI - Quantitative detection of promoter hypermethylation of multiple genes in the tumor, urine, and serum DNA of patients with renal cancer. PMID- 15879776 TI - Serum protein profiling by SELDI mass spectrometry: detection of multiple variants of serum amyloid alpha in renal cancer patients. PMID- 15879777 TI - The level of cadherin-6 mRNA in peripheral blood is associated with the site of metastasis and with the subsequent occurrence of metastases in renal cell carcinoma. PMID- 15879778 TI - Small cell carcinoma of the urinary bladder. A clinicopathologic analysis of 64 patients. PMID- 15879779 TI - Her-2/neu protein overexpression and gene amplification in human transitional cell carcinoma of the bladder. PMID- 15879782 TI - The effect of statins on serum prostate specific antigen levels in a cohort of airline pilots: a preliminary report. AB - PURPOSE: Reports of the effect of treatment with statins on prostate cancer risk are inconsistent. We performed a pilot study to assess the effect of statin treatment on a surrogate marker for prostate cancer risk, that is serum prostate specific antigen (PSA), in a cohort of airline pilots from 1992 to 2001. MATERIALS AND METHODS: Subject medical records were abstracted for data on age, PSA testing, hyperlipidemia and treatment with statins. The treatment group was composed of 15 men with hypercholesterolemia who received statins and the comparison group of 85 with normal serum lipid levels during the review period. The mean +/- SD and the Wilcoxon rank sum test were used for analyses. RESULTS: Serum PSA was significantly higher in the treatment group at baseline relative to the comparison group (p = 0.05). Interestingly there was no significant difference between the groups on subsequent followup. There was a 41.6% decrease in mean serum PSA in the treated group by visit 4. Simultaneously mean serum PSA increased by 38% in the untreated group. CONCLUSIONS: Our results suggest that treatment with statins may lower serum PSA with time. These results must be confirmed in a larger study population while controlling for potential confounders. If validated, our observation provides a rationale for further studies of the role of this class of drugs for prostate cancer chemoprevention. PMID- 15879783 TI - Prostate cancer detection in men with prostate specific antigen 4 to 10 ng/ml using a combined approach of contrast enhanced color Doppler targeted and systematic biopsy. AB - PURPOSE: Transrectal gray scale ultrasound guided biopsy is the standard method for diagnosing prostate cancer (PC). Improved cancer detection with ultrasound contrast agents is related to better detection of tumor vascularity. We evaluated the impact of a combined approach of contrast enhanced, color Doppler targeted biopsy (CECD) and systematic biopsy (SB) for the PC detection rate in men with prostate specific antigen (PSA) 4.0 to 10 ng/ml. MATERIALS AND METHODS: We examined 380 screening volunteers with a total PSA of 4.0 to 10 ng/ml (percent free PSA less than 18). CECD was always performed before SB. Another investigator blinded to contrast enhanced findings performed 10 SBs. The cancer detection rate for the CECD, SB and combined approaches was assessed. RESULTS: PC was detected in 143 of 380 patients (37.6%, mean total PSA 6.2 ng/ml). The PC detection rate for CECD and for SB was 27.4% and 27.6%, respectively. The overall cancer detection rate with the 2 methods combined was 37.6%. For targeted biopsy cores the detection rate was significantly better than for SB cores (32.6% vs 17.9%, p <0.01). CECD in a patient with cancer was 3.1-fold more likely to detect PC than SB. CONCLUSIONS: CECD allows for the detection of lesions that cannot be found on gray scale ultrasound or SB. CECD allows for assessment of neovascularity associated with PC. However, the combined use of CECD and SB allows for maximal detection of PC with a detection rate of 37.6% in our patients with PSA 4 to 10 ng/ml. PMID- 15879784 TI - Development and validation of a nomogram predicting the outcome of prostate biopsy based on patient age, digital rectal examination and serum prostate specific antigen. AB - PURPOSE: We developed and validated a nomogram which predicts presence of prostate cancer (PCa) on needle biopsy. MATERIALS AND METHODS: We used 3 cohorts of men who were evaluated with sextant biopsy of the prostate and whose presenting prostate specific antigen (PSA) was not greater than 50 ng/ml. Data from 4,193 men from Montreal, Canada were used to develop a nomogram based on age, digital rectal examination (DRE) and serum PSA. External validation was performed on 1,762 men from Hamburg, Germany. Data from these men were subsequently used to develop a second nomogram in which percent free PSA (%fPSA) was added as a predictor. External validation was performed using 514 men from Montreal. Both nomograms were based on multivariate logistic regression models. Predictive accuracy was evaluated with areas under the receiver operating characteristic curve and graphically with loess smoothing plots. RESULTS: PCa was detected in 1,477 (35.2%) men from Montreal, 739 (41.9%) men from Hamburg and 189 (36.8%) men from Montreal. In all models all predictors were significant at 0.05. Using age, DRE and PSA external validation AUC was 0.69. Using age, DRE, PSA and %fPSA external validation AUC was 0.77. CONCLUSIONS: A nomogram based on age, DRE, PSA and %fPSA can highly accurately predict the outcome of prostate biopsy in men at risk for PCa. PMID- 15879785 TI - Testosterone as a predictor of pathological stage in clinically localized prostate cancer. AB - PURPOSE: Substantial controversy exists in the literature regarding the association between pretreatment testosterone and disease outcome in patients with prostate cancer. We explored the relationship between preoperative total testosterone, and pathological stage and progression in patients with clinically localized prostate cancer treated with radical prostatectomy. MATERIALS AND METHODS: We retrospectively reviewed the records of consecutive patients with clinically localized prostate cancer treated with radical prostatectomy between January 1990 and June 2003. A total of 326 patients with pretreatment testosterone levels available were eligible for this analysis. Biochemical progression (BCR) was defined by postoperative prostate specific antigen (PSA) greater than 0.4 ng/ml with a confirmatory increase and it occurred in 41 men. No men received adjuvant therapy. Univariate and multivariate logistic regression analyses were done to examine whether pretreatment testosterone was associated with pathological stage. Cox regression was used to assess the association of testosterone and BCR. RESULTS: Median PSA was 6.01 ng/ml (range 0.13 to 86), testosterone was 385 ng/dl (range 133 to 998) and followup was 36 months (range 4 to 136). In 245 patients (75%) disease was organ confined. Lower testosterone correlated with adverse pathological stage on multivariate analysis (p = 0.01), as did clinical stage, biopsy grade and PSA. However, we found no relationship between testosterone and BCR after adjusting for covariates. Furthermore, we found no evidence of an interaction between PSA and testosterone (p = 0.4). CONCLUSIONS: On multivariate analysis low preoperative total testosterone was associated with advanced pathological stage but not with BCR. Future studies are warranted with data on more patients who have progressed. PMID- 15879786 TI - The University of California, San Francisco Cancer of the Prostate Risk Assessment score: a straightforward and reliable preoperative predictor of disease recurrence after radical prostatectomy. AB - PURPOSE: Multivariate prognostic instruments aim to predict risk of recurrence among patients with localized prostate cancer. We devised a novel risk assessment tool which would be a strong predictor of outcome across various levels of risk, and which could be easily applied and intuitively understood. MATERIALS AND METHODS: We studied 1,439 men diagnosed between 1992 and 2001 who had undergone radical prostatectomy and were followed in the Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE) database, a longitudinal, community based disease registry of patients with prostate cancer. Disease recurrence was defined as prostate specific antigen (PSA) 0.2 ng/ml or greater on 2 consecutive occasions following prostatectomy or a second cancer treatment more than 6 months after surgery. The University of California, San Francisco-Cancer of the Prostate Risk Assessment (UCSF-CAPRA) score was developed using preoperative PSA, Gleason score, clinical T stage, biopsy results and age. The index was developed and validated using Cox proportional hazards and life table analyses. RESULTS: A total of 210 patients (15%) had recurrence, 145 by PSA criteria and 65 by second treatment. Based on the results of the Cox analysis, points were assigned based on PSA (0 to 4 points), Gleason score (0 to 3), T stage (0 to 1), age (0 to 1) and percent of biopsy positive cores (0 to 1). The UCSF-CAPRA score range is 0 to 10, with roughly double the risk of recurrence for each 2-point increase in score. Recurrence-free survival at 5 years ranged from 85% for a UCSF-CAPRA score of 0 to 1 (95% CI 73%-92%) to 8% for a score of 7 to 10 (95% CI 0%-28%). The concordance index for the UCSF-CAPRA score was 0.66. CONCLUSIONS: The UCSF-CAPRA score is a straightforward yet powerful preoperative risk assessment tool. It must be externally validated in future studies. PMID- 15879787 TI - Detection of early lymph node metastases in prostate cancer by laparoscopic radioisotope guided sentinel lymph node dissection. AB - PURPOSE: Radioisotope guided sentinel lymph node (SLN) dissection (SLND) for prostate cancer has been shown to increase the sensitivity of detecting early metastases in open pelvic lymph node dissection. We developed a technique that allows SLND to be performed by laparoscopy in conjunction with laparoscopic radical prostatectomy. MATERIALS AND METHODS: In 71 consecutive patients SLND was performed by 1 surgeon preceding laparoscopic radical prostatectomy. Mean preoperative prostate specific antigen was 8.88 ng/ml (range 2.1 to 25.4). At 24 hours prior to surgery 3 ml (200 MBq) Tc labeled human albumin colloid were injected into the prostate gland under transrectal ultrasound guidance. An especially designed laparoscopic gamma probe was used to measure radioactivity during surgery. SLNs were identified and removed. If frozen section analysis showed metastases, extended pelvic lymph node dissection was performed. RESULTS: Radioactivity was detected on 2, 1 and no sides in 50 (70.4%), 19 (26.7%) and 2 patients (2.8%), respectively. In 81 of the 142 pelvic side walls (54.7%) SLNs were exclusively outside of the obturator fossa. Histopathological examination showed metastases to SLNs in 9 patients (12.9%). Eight of the 11 detected metastases (72.7%) were outside of the obturator fossa. Lymph node metastases were exclusively found in Tc marked lymph nodes. Mean tumor size was 1.7 mm (range 0.2 to 3.9). CONCLUSIONS: SLND is feasible by laparoscopy. It detects micrometastases outside of the obturator fossa in a significant number of patients. We noted that the transperitoneal approach allowing wide exposure and a gamma probe with a 90-degree lateral energy window is the most important factor to enable successful laparoscopic SLND. PMID- 15879788 TI - Long-term outcome for men with androgen independent prostate cancer treated with ketoconazole and hydrocortisone. AB - PURPOSE: The combination of high dose ketoconazole and hydrocortisone (HDK) is active against androgen independent prostate cancer (AIPC). Median response times with HDK tend to be brief but a significant minority of AIPC patients benefit with extended responses. Well characterized response and survival information, especially in the cohort of patients who experience these longer, more durable, responses has not been previously reported. Characterization of this subgroup is of particular interest since men with long-term responses derive the greatest benefit from HDK therapy. MATERIALS AND METHODS: The medical records of 78 patients with AIPC treated with HDK between March 1991 and February 1999 were retrospectively reviewed. Baseline clinical and laboratory factors predictive of prolonged response and survival were identified. RESULTS: The median baseline prostate specific antigen (PSA) before the initiation of HDK was 25.1. The number of patients with zero, 1 to 3, and more than 3 lesions on bone scan were 25, 35 and 18, respectively. Median and mean time to PSA progression was 6.7 and 14.5 months. Median and mean survival time was 38.0 and 42.4 months, respectively. Response time and survival were highly correlated (r = 0.799). A total of 34 (44%) men had a greater than 75% decrease in PSA. The median survival times in men with more vs less than a 75% decrease were 60 vs 24 months, respectively. In a Cox proportional hazard regression, prolonged survival was predicted by percent PSA decrease, extent of disease on bone scan and baseline PSA. CONCLUSIONS: Ketoconazole can induce prolonged responses, occasionally lasting for years. Long responses are more likely to occur in men initiating HDK earlier in the course of disease before the cancer burden becomes excessive. PMID- 15879789 TI - Urinary fistulas following external radiation or permanent brachytherapy for the treatment of prostate cancer. AB - PURPOSE: Prostate cancer (PCa) radiotherapy (RT), including brachytherapy, may lead to significant morbidity, including urinary fistulas. If conservative measures fail, urinary and/or fecal diversion is often required. In this study we examined a series of patients with fistulas that developed after pelvic radiation therapy and explored potential predisposing factors and treatment recommendations for refractory fistulas. MATERIALS AND METHODS: Patients were identified who received radiation therapy for PCa between 1977 and 2002, and subsequently had a fistula to the urinary tract. Patients were excluded who had diverticulitis, inflammatory bowel disease, a history of recent radical retropubic prostatectomy (possible iatrogenic etiology) or cancer in the excised fistula. Data were extracted from patient charts, mailed questionnaires and outside records. RESULTS: A total of 51 patients were identified with a history of radiation for PCa who subsequently had a urinary fistula. Of 20 patients meeting inclusion criteria 30% received external beam RT alone, 30% received brachytherapy and 40% received combined external beam RT/brachytherapy. Most fistulas (80%) were from the rectum to the urinary tract with an average diameter of 3.2 cm. Of patients with rectal fistulas 81% had a history of rectal stricture, urethral stricture, rectal biopsy, rectal argon beam therapy or transurethral prostate resection after radiation. All patients with rectourethral fistulas who achieved symptomatic resolution required urinary and fecal diversion. CONCLUSIONS: Conservative treatment is generally ineffective in the management of large urinary fistulas. Surgical intervention offers symptomatic relief and improved quality of life in most patients. PMID- 15879790 TI - Melanoma of the penis, scrotum and male urethra: a 40-year single institution experience. AB - PURPOSE: Genitourinary melanoma is rare and classically associated with a poor prognosis. We describe our experience with 10 patients with penile or urethral involvement. In addition, we present what is to our knowledge the largest reported series of melanoma of the scrotum (6 cases). MATERIALS AND METHODS: We reviewed the records of 16 men who presented consecutively to our institution with genitourinary melanoma between 1962 and 2000. Clinical and pathological characteristics were assessed, including Breslow thickness, primary surgical intervention and clinical course. RESULTS: Of 10 patients with penile or urethral melanoma 1997 American Joint Committee on Cancer melanoma pathological stage was T1 (depth less than 0.75 mm) in 4, T2 (0.75 to 1.5 mm) in 3 and T3 (1.51 to 4 mm) in 3. Only 1 of 4 patients with clinically palpable inguinal nodes had inguinal metastases at lymphadenectomy (BILND) and 3 who underwent prophylactic superficial BILND had negative findings. In 7 patients with T1-2N0M0 disease there were no local recurrences after wide local excision (WLE) or partial penectomy at a median followup of 35 months. Six of 7 men were rendered disease free. One patient died of melanoma that developed at a second primary site. The 3 patients with T3 tumors who underwent partial (2) or radical (1) penectomy with or without BILND died of disease (2) or had progression (1). In all patients with penile melanoma the 5-year actuarial disease specific and recurrence-free survival rates were 80% and 60%, respectively, at a median followup of 39 months (range 20 to 210). Six patients with scrotal melanoma were treated with WLE without local recurrences. Three of the 6 patients had palpable inguinal nodes, of whom 2 died after chemotherapy for unresectable disease and 1 died of other causes 51 months after negative BILND. The 3 men with clinically negative groins who did not undergo prophylactic BILND had distant (1) or regional (2) metastases and died of disease. In patients with scrotal melanoma the 5-year actuarial disease specific and recurrence-free survival rates were 33.3% and 33.3%, respectively, at a median followup of 36 months. CONCLUSIONS: Partial penectomy or WLE provided effective local control for low stage penile or urethral melanomas and all scrotal lesions. Patients showing clinically positive, proven metastasis died despite appropriate surgical procedures and multi-agent chemotherapy. Prophylactic modified inguinal lymphadenectomy should be considered in select patients with penile, scrotal and anterior urethral melanoma. PMID- 15879791 TI - Prospective, multicenter, randomized phase II trial of the herbal supplement, PC SPES, and diethylstilbestrol in patients with androgen-independent prostate cancer. PMID- 15879792 TI - American Society of Clinical Oncology recommendations for the initial hormonal management of androgen-sensitive metastatic, recurrent, or progressive prostate cancer. PMID- 15879793 TI - High level of androgen receptor is associated with aggressive clinicopathologic features and decreased biochemical recurrence-free survival in prostate. Cancer patients treated with radical prostatectomy. PMID- 15879794 TI - Propionibacterium acnes associated with inflammation in radical prostatectomy specimens: a possible link to cancer evolution? AB - PURPOSE: Inflammation is commonly observed in the prostate gland and has been implicated in the development of prostate cancer. The etiology of prostatic inflammation is unknown. However, the involvement of a carcinogenic infectious agent has been suggested. MATERIALS AND METHODS: Prostatic tissue from 34 consecutive patients with prostate cancer was cultured to detect the presence of bacterial agents. Prostatic inflammation was assessed by histological examination of wholemount tissue sections. RESULTS: The predominant microorganism detected was Propionibacterium acnes, found in 35% of prostate samples. A significantly higher degree of prostatic inflammation was observed in cases culture positive for P. acnes (p =0.007). P. acnes was separated into 3 groups based on cell surface properties, phenotype and genetic grouping. All skin control isolates were classified as group 1 whereas most prostatic isolates were classified as groups 2 and 3. CONCLUSIONS: P. acnes has been isolated from prostatic tissues in men who underwent radical prostatectomy for localized cancer and has been shown to be positively associated with prostatic inflammation. This inflammation may then be linked to the evolution of carcinoma. Furthermore, organisms infecting these patients with prostate cancer differ genetically and phenotypically from the commonly identified cutaneous P. acnes isolates, suggesting that specific subtypes may be involved in development of prostatic inflammation. PMID- 15879795 TI - Hyperbaric oxygen for the treatment of fournier's gangrene. AB - PURPOSE: Fournier's gangrene is a necrotizing fasciitis of the genitalia that is associated with high morbidity and mortality. Groups at many institutions have initiated routine adjuvant hyperbaric oxygen (HBO) therapy. We examined whether HBO has made a difference in the morbidity, mortality and costs associated with treating this disease. We also analyzed predictors of extended hospital stay and mortality. MATERIALS AND METHODS: The records of patients with the hospital discharge diagnoses of Fournier's gangrene, necrotizing fasciitis, gangrene of the genitalia and scrotal gangrene from 1993 to 2002 were reviewed. Data concerning clinical presentation characteristics, hospital stay, complications, hospital charges and outcomes, including graft failure and death, were analyzed. RESULTS: A total of 42 patients were identified and followed a median 4.2 years. Of the patients 16 underwent surgical debridement and antibiotic therapy alone, and 26 were treated with HBO plus surgery and antibiotics. Overall disease specific mortality was 21.4%, that is 12.5% in the nonHBO group and 26.9% in the HBO group. Three or more complications occurred in 13% of nonHBO and in 19% of HBO cases, of which the most common was myocardial infarction. The skin graft failure rate was 6% (nonHBO) and 8% (HBO). Physical disability was a statistically significant predictor of extended hospital stay (p <0.01). There was a trend toward a correlation between known coronary artery disease and death (p = 0.2). A statistically significant difference was noted in average daily hospital charges in nonHBO vs HBO cases ($2,552 vs $3,384 daily, p <0.01). CONCLUSIONS: These data do not support routine HBO in the treatment of Fournier's gangrene. There was a trend toward higher morbidity and mortality in the HBO group, suggesting that treatment may have been given to patients who were more ill. PMID- 15879796 TI - Coccidioidomycosis of the male genital tract. AB - PURPOSE: Symptomatic genital tract infection is a rare manifestation of disseminated coccidioidomycosis. We characterized the clinical presentation, diagnosis, management and outcome in male patients with genitourinary coccidioidomycosis. MATERIALS AND METHODS: We reviewed all cases of genital tract coccidioidomycosis involving testes, epididymis or prostate at our institution between 1990 and 2000, and searched the medical literature for all reports since the first reported case in 1943. RESULTS: A total of 30 male patients with a median age of 58 years who had coccidioidomycosis of the genital tract were identified, including 6 at our institution. Four patients (13%) had a simultaneous pulmonary infection and 63% (19) had a remote history of primary pulmonary coccidioidomycosis. The most commonly involved genital tract sites were the epididymis in 18 cases, prostate in 14 and testes in 6. Patients with prostatitis presented with urinary obstruction, prostatic enlargement, tenderness and palpable nodules. Most patients with epididymal infection presented with scrotal swelling, tenderness and induration. All 30 patients (100%) had histopathological evidence of granulomatous inflammation with fungal spherules. Urine fungal cultures were positive in 19 cases (63%). A total of 12 patients received combined medical and surgical treatment, and 18 underwent surgical excision only. Most immunocompetent patients with isolated genital coccidioidomycosis did well with surgical resection alone. Six deaths occurred in patients with multifocal, extragenital, disseminated disease. CONCLUSIONS: Male genitourinary coccidioidomycosis is rare but it should be considered in the differential diagnosis of patients with exposure to the endemic area who present with prostatitis or epididymitis. PMID- 15879797 TI - Psychophysical evidence of hypersensitivity in subjects with interstitial cystitis. AB - PURPOSE: We quantified differences in somatic and visceral sensation in healthy subjects and subjects with interstitial cystitis (IC). MATERIALS AND METHODS: A total of 13 subjects with IC and 13 healthy subjects answered psychological questionnaires and underwent psychophysical testing of thermal and pressure thresholds for sensation as well as the ischemic forearm test of pain tolerance. A subset of subjects also underwent bladder sensory testing with the determination of 3 consecutive cystometrograms. Ratings of intensity and unpleasantness were determined. RESULTS: Subjects with IC were significantly more sensitive to deep tissue measures of sensation related to pressure, ischemia and bladder than healthy subjects. Cutaneous thermal pain measures were similar in the 2 groups. Psychological measures indicated higher reactivity in subjects with IC. CONCLUSIONS: Similar to other visceral pain disorders, such as irritable bowel syndrome, hypersensitivity to somatic stimuli was noted in subjects with IC. This suggests altered central mechanisms in the processing of sensory events from the bladder. PMID- 15879798 TI - Effects of fluoroquinolones on the migration of human phagocytes through Chlamydia pneumoniae-infected and tumor necrosis factor alpha-stimulated endothelial cells. PMID- 15879799 TI - Cranberries for preventing urinary tract infections. PMID- 15879800 TI - Comparison of first void urine and urogenital swab specimens for detection of Mycoplasma genitalium and Chlamydia trachomatis by polymerase chain reaction in patients attending a sexually transmitted disease clinic. PMID- 15879801 TI - Comparison of three nucleic acid amplification tests for detection of Chlamydia trachomatis in urine specimens. PMID- 15879802 TI - Fixed drug eruption--a sexually inducible reaction? PMID- 15879803 TI - Chapter 1: AUA guideline on management of staghorn calculi: diagnosis and treatment recommendations. PMID- 15879804 TI - Ethnic background has minimal impact on the etiology of nephrolithiasis. AB - PURPOSE: Nephrolithiasis disproportionately affects white patients. However, recent studies propose an increase in the incidence of stone disease in nonwhite populations. We compared the metabolic risk factors of ethnically disparate stone formers from the same geographic region. MATERIALS AND METHODS: A retrospective review of 1,141 patients identified 98 (9%) nonwhite stone formers. Of these individuals 60 underwent a comprehensive metabolic evaluation, comprising 44 black, 8 Asian and 8 Hispanic patients. A similar sex and age matched group of 66 white stone forming patients were also identified for comparative analysis. Stone analyses were recorded when available. Urinary metabolic abnormalities were defined as low urine volume-urine volume less than 2,000 cc, gouty diathesis-pH 5.5 or less (normal level 5.5 to 6.5), hypercalciuria-calcium greater than 200 mg, hyperoxaluria-oxalate greater than 45 mg, hyperuricosuria-uric acid greater than 600 mg, hypocitraturia-citrate less than 600 mg and purine gluttony-sulfate greater than 20 mg. RESULTS: The incidence of metabolic abnormalities was surprisingly similar between the white and nonwhite stone formers. Whites have a higher prevalence of hypercalciuria compared with nonwhites (67% vs 25%, respectively, p <0.01). This comparison persisted when the white group was compared with individual ethnic groups (25% in each group). Whites also displayed a higher mean urinary calcium level (233 mg) than their nonwhite counterparts overall (146 mg), specifically with respect to blacks (146 mg, p <0.01). Asians had higher urine volumes with respect to whites and blacks (p <0.01) and, therefore, a decreased prevalence of low urine volumes (37.5% vs 74.2% and 79.5%, respectively). Hypocitraturia, hyperuricosuria, hyperoxaluria, gouty diathesis and high sulfate levels were equally represented among all ethnic groups. CONCLUSIONS: Although there appears to be a predominance of stone disease among whites, all racial groups demonstrated a remarkable similarity in the incidence of underlying metabolic abnormalities. These results suggest that dietary and environmental factors may be as important as ethnicity in the etiology of stone disease. PMID- 15879805 TI - Prospective, randomized trial comparing shock wave lithotripsy and ureteroscopy for lower pole caliceal calculi 1 cm or less. AB - PURPOSE: The optimal management of lower pole renal calculi is controversial. We compared shock wave lithotripsy (SWL) and ureteroscopy (URS) for the treatment of patients with small lower pole stones in a prospective, randomized, multicenter trial. MATERIALS AND METHODS: A total of 78 patients with 1 cm or less isolated lower pole stones were randomized to SWL or URS. The primary outcome measure was stone-free rate on noncontrast computerized tomography at 3 months. Secondary outcome parameters were length of stay, complication rates, need for secondary procedures and patient derived quality of life measures. RESULTS: A total of 67 patients randomized to SWL (32) or URS (35) completed treatment. The 2 groups were comparable with respect to age, sex, body mass index, side treated and stone surface area. Operative time was significantly shorter for SWL than URS (66 vs 90 minutes). At 3 months of followup 26 and 32 patients who underwent SWL and URS had radiographic followup that demonstrated a stone-free rate of 35% and 50%, respectively (p not significant). Intraoperative complications occurred in 1 SWL case (unable to target stone) and in 7 URS cases (failed access in 5 and perforation in 2), while postoperative complications occurred in 7 SWL and 7 URS cases. Patient derived quality of life measures favored SWL. CONCLUSIONS: This study failed to demonstrate a statistically significant difference in stone-free rates between SWL and URS for the treatment of small lower pole renal calculi. However, SWL was associated with greater patient acceptance and shorter convalescence. PMID- 15879806 TI - The comparison and efficacy of 3 different alpha1-adrenergic blockers for distal ureteral stones. AB - PURPOSE: alpha1-Adrenergic blockers have recently been shown to increase the rate of spontaneous passage of distal ureteral stones. We compared efficacy of 3 different alpha1-adrenergic blockers for this purpose. MATERIALS AND METHODS: A total of 114 patients between 18 and 65 years old who had lower ureteral stones were included in the study. Patients were randomly divided into 4 groups. Group 1 consisted of 28 patients and acted as the control group. Group 2 comprised 29 patients who received tamsulosin, group 3 was 28 patients receiving terazosin and group 4 was 29 patients receiving doxazosin. These agents were given for up to a month and hydration was also recommended simultaneously. Every week patients were controlled with x-rays of the kidneys, ureters, bladder and urinary ultrasonography. Meanwhile the number of pain episodes, analgesic dosage and the number of days for spontaneous passage of the calculi through the ureter were also recorded. RESULTS: There were no differences between the groups with respect to age, weight, height, sex and stone size. The calculi passed through the ureter spontaneously in 15 patients in group 1 (53.57%), in 23 patients in group 2 (79.31%), in 22 patients in group 3 (78.57%), and in 22 patients in group 4 (75.86%). In groups 2 to 4 the number of pain episodes, expulsion time and analgesic dosage were found to be lower compared with those in group 1. CONCLUSIONS: alpha1-Adrenergic blockers increase the frequency of spontaneous passage of the distal ureteral calculi. All 3 agents tested were equally efficacious. PMID- 15879807 TI - Is newer always better? A comparative study of 3 lithotriptor generations. AB - PURPOSE: At a single center we compared the efficacy of 3 generations of lithotriptors using identical protocol inclusion and followup criteria but with different modes of anesthesia. MATERIALS AND METHODS: We compared stone disintegration and dilatation of the pyelocaliceal system achieved in a prospective, randomized trial comparing the original HM3 (Dornier Medtech, Kennesaw, Georgia) and Lithostar Plus (LSP) lithotriptors, and a matched, consecutive series of 107 treatments with the Modulith SLX. Stone disintegration and dilatation of the pyelocaliceal system were evaluated by abdominal plain x ray and renal ultrasonography 1 day and 3 months after treatment. RESULTS: A total of 82 treatments with the HM3, 75 with the LSP and 107 with the SLX were analyzed, matched for stone burden and location within the pyelocaliceal system. On postoperative day 1, 91%, 65% and 48% patients treated with the HM3, LSP and SLX, respectively, were stone-free or had fragments that were 2 mm or less (HM3 vs LSP p <0.001, HM3 vs SLX p <0.001 and LSP vs SLX p = 0.015). Three to 5 mm fragments were found in 7%, 21% and 35% of patients (p = 0.006, <0.001 and 0.06), and fragments 6 mm or greater were found in 1%, 14% and 15% (p = 0.002, <0.001 and 0.1, respectively). The re-treatment rate was 4% in the HM3 group, 13% in the LSP group and 38% in the SLX group (HM3 vs LSP p = 0.05, HM3 vs SLX p <0.001 and LSP vs SLX p <0.001). Obstructive pyelonephritis occurred in 1% of the HM3 group, 8% of the LSP group and 5% of the SLX group (HM3 vs LSP p = 0.02, HM3 vs SLX p = 0.12 and LSP vs SLX p = 0.4). All re-treatments except those in 5 patients were performed with the HM3. Therefore, the 3-month stone-free rate was comparable in all 3 groups (HM3 87%, LSP 80% and SLX 81%). CONCLUSIONS: This study indicates that the HM3 lithotriptor disintegrates caliceal and renal pelvic stones better than the LSP and SLX machines, resulting in fewer complications and re treatments. Disintegration with the LSP machine was also superior to that of the SLX with a need for fewer re-treatments. PMID- 15879808 TI - Comparison of flexible ureteroscopes: deflection, irrigant flow and optical characteristics. AB - PURPOSE: We measured and compared the deflection, irrigation flow rates, distortion, resolution and light transmission of new generation flexible ureteroscopes. MATERIALS AND METHODS: Multiple characteristics of 5 flexible ureteroscopes (ACMI DUR-8 Elite, Olympus URF-P3, Storz 11278AU1 [Flex-X], Wolf 7330.072 and Wolf 7325.172) commonly available in the market were measured and compared. Measured data included active deflection, irrigation flow rates and optical characteristics. Each ureteroscope was evaluated with an empty working channel and with various accessories. Optical characteristics, specifically resolution and distortion, were measured using test targets (Edmund Optics, Barrington, New Jersey). Light transmission was also measured from the ureteroscope tip at 50% and 100% intensity. All 5 flexible ureteroscopes were tested in a laboratory setting using a Storz OR 1 system to capture the images. RESULTS: For all 5 ureteroscopes the angle of deflection was most impaired by a 365 microm laser fiber probe and least impaired by a 2.2Fr nitinol basket. Among all 5 ureteroscopes irrigation flow rate was most impaired with a 3.0Fr basket and least impaired with 200 microm laser fiber. The Wolf 7325.172 had the highest observed resolution of 25.39 lines per mm and the Wolf 7330.072 had the lowest distortion at 11.9%. The Karl Storz Flex-X and the ACMI DUR-8 Elite had the highest light output at 374 and 364 mV, respectively. CONCLUSIONS: The various flexible ureteroscopes differ with regard to flow rates as well as degree of deflection with either an empty or an occupied working channel. The Wolf flexible ureteroscope with a slightly larger working channel and a fused quartz bundle provided for superior flow and better optical performance. However, the greatest amount of tip deflection and highest light output were found in the ACMI and Karl Storz flexible ureteroscopes. PMID- 15879809 TI - Ureteroscopic lithotripsy under local anesthesia: analysis of the effectiveness and patient tolerability. PMID- 15879810 TI - Ureteric stenting after ureteroscopy for ureteric stones: a prospective randomized study assessing symptoms and complications. PMID- 15879812 TI - Comparison of open versus laparoscopic pyeloplasty techniques in treatment of uretero-pelvic junction obstruction. PMID- 15879813 TI - Changes of renal blood flow in nephroptosis: assessment by color Doppler imaging, isotope renography and correlation with clinical outcome after laparoscopic nephropexy. PMID- 15879814 TI - The bladder does not appear to have a dynamic secreted continuous mucous gel layer. AB - PURPOSE: We determined whether the nature of any protective barrier in the bladder is composed of a secreted mucous gel layer. MATERIALS AND METHODS: We collected 24-hour urine samples for analysis from 8 healthy 22 to 49-year-old volunteers and 5, 19 to 59-year-old patients treated with bladder reconstruction, in addition to scrapings from 100 freshly slaughtered pig bladders. Samples were subjected to homogenization, dialysis, freeze-drying, papain digestion, gel chromatography, equilibrium density gradient centrifugation, periodic acid-Schiff assay and amino acid analysis. Normal human bladder, pig bladder, normal ileum and transposed intestinal segments were studied for the presence of a mucous layer using a new method of histological analysis. RESULTS: Mucin content in normal urine is 2.7 mg/24 hours, meaning that less than 0.6% of nondialyzable material in normal urine is mucin. The mucin content of urine from reconstructed bladders amounted to 86 mg/24 hours (5.2% of nondialyzable material). We observed that glycosaminoglycans accounted for 41% of the peak total elution volume of PAS positive material in normal urine. Mucin estimation in urine can be grossly overestimated if contaminating glycoconjugates are not removed. Biochemical analysis of material scraped off the pig bladder surface demonstrated that the maximum thickness of a continuous layer that could be achieved was 13.6 mum. While we could visualize an obvious mucous layer on control ileal samples and biopsies of transposed ileal segments from patients with bladder reconstruction, we were unable to note a distinct, measurable mucous layer lining the bladder surface in humans or pigs. CONCLUSIONS: Mucin levels in normal human and pig urine would be enough for slow turnover of a thin barrier but the large increase in mucin in the urine of patients with transposed intestinal segments demonstrates that any layer in normal bladder is much different than that lining the transposed intestinal segment. The most likely constituents of this barrier are membrane bound rather than secreted mucins along with the proteoglycan components of the glycocalix. PMID- 15879815 TI - Heroic measures may not always be justified in extensive urethral stricture due to lichen sclerosus (balanitis xerotica obliterans). PMID- 15879816 TI - Long-term results of a self-expanding wallstent in the treatment of urethral stricture. PMID- 15879817 TI - Early and late treatment-related morbidity following radical cystectomy. PMID- 15879819 TI - Urinary conduit formation using retubularized bowel from continent urinary diversion or intestinal augmentations: I. A multi-institutional experience. PMID- 15879820 TI - Treatment of neurogenic detrusor overactivity in spinal cord injured patients by conditional electrical stimulation. AB - PURPOSE: The feasibility of automatic event driven electrical stimulation of the dorsal penile/clitoral nerve in the treatment of neurogenic detrusor overactivity (NDO) was evaluated in individuals with spinal cord injury. MATERIALS AND METHODS: The study included 2 women and 14 men older than 18 years with NDO, bladder capacity below 500 ml and complete or incomplete suprasacral spinal cord injury. Detrusor pressure (Pdet) was recorded during ordinary, natural bladder filling. In a similar subsequent recording Pdet was used to trigger electrical stimulation when pressure exceeded 10 cm H2O. RESULTS: Of the 16 patients enrolled in this study 13 had increased bladder capacity together with a storage pressure decrease as a result of automatic, event driven electrical stimulation. In 2 patients stimulation could not inhibit the first undesired contraction, leakage occurred and finally 1 could not tolerate stimulation. During stimulated filling Pdet never exceeded 55 cm H2O. Thus, storage pressure was sufficiently low to prevent kidney damage. An average bladder capacity increase of 53% was achieved. CONCLUSIONS: This study demonstrates the feasibility of automatic, event driven electrical stimulation in the treatment of NDO. Although the setup in this experiment is not suitable in a clinical setting, the treatment modality is promising and it warrants further investigation. PMID- 15879821 TI - Can a baseline prostate specific antigen level identify men who will have lower urinary tract symptoms later in life? AB - PURPOSE: We evaluated the relationship between baseline prostate specific antigen (PSA) and subsequent lower urinary tract symptom development during 3 decades in unselected men in the Baltimore Longitudinal Study of Aging. MATERIALS AND METHODS: Urinary questionnaires were used to evaluate lower urinary tract symptoms in 704 men during 3 decades. The number of repeat evaluations was 1 to 18. We divided subjects into age groups of younger than 50 and 50 to 69.9 years at the time of the first PSA evaluation. Subjects were divided into 3 PSA groups based on initial PSA below the 25th, 25th to 75th and above the 75th percentile. A mixed effects Poisson model was used to test whether there was a significant relationship between PSA grouping and symptom score with time. RESULTS: There was no statistically significant difference in symptom score distribution across PSA percentiles in men younger than 50 years (p = 0.87) or 50 to 69.9 years old (p = 0.59). When age was used as an independent variable in the model, there was no statistically significant relationship between baseline PSA and symptom score (p = 0.38). CONCLUSIONS: These data suggest that PSA is not a useful predictor of the development of lower urinary tract symptoms in unselected, asymptomatic men. PMID- 15879822 TI - A prospective, randomized, placebo controlled, double-blind study of pelvic electromagnetic therapy for the treatment of chronic pelvic pain syndrome with 1 year of followup. AB - PURPOSE: Male chronic pelvic pain syndrome is a condition of uncertain etiology and treatment is often unsatisfactory. There is evidence that the symptom complex may result from pelvic floor muscular dysfunction and/or neural hypersensitivity/inflammation. We hypothesized that the application of electromagnetic therapy may have a neuromodulating effect on pelvic floor spasm and neural hypersensitivity. MATERIALS AND METHODS: Following full Stamey localization men with National Institute of Diabetes and Digestive and Kidney Diseases category III prostatitis were prospectively randomized to receive active electromagnetic or placebo therapy. Active therapy consisted of 15 minutes of pelvic floor stimulation at a frequency of 10 Hz, followed by a further 15 minutes at 50 Hz, twice weekly for 4 weeks. Patients were evaluated at baseline, 3 months and 1 year after treatment using validated visual analog scores. RESULTS: A total of 21 men with a mean age of 47.8 years (range 25 to 67) were analyzed. Mean symptom scores decreased significantly in the actively treated group at 3 months and 1 year (p <0.05), unlike the placebo group, which showed no significant change (p >0.05). Subanalysis of those receiving active treatment showed that the greatest improvement was in pain related symptoms. CONCLUSIONS: The novel use of pelvic floor electromagnetic therapy may be a promising new noninvasive option for chronic pelvic pain syndrome in men. PMID- 15879823 TI - A population based study of incidence and treatment of benign prostatic hyperplasia among residents of Olmsted County, Minnesota: 1987 to 1997. AB - PURPOSE: The treatment of benign prostatic hyperplasia (BPH), a common problem faced by aging men, has changed dramatically during the last decade. While BPH has long been considered a surgical condition, the advent of less invasive therapies makes the evaluation of the use of these various treatments important. Thus, this population based study was undertaken to describe the incidence and treatment for BPH from 1987 to 1997 among residents of Olmsted County, Minnesota. MATERIALS AND METHODS: We identified all Olmsted County men with a new diagnosis of BPH from 1987 to 1997 through the Medical and Surgical Indices made available through the Rochester Epidemiology Project. Overall year and age specific incidence rates were calculated for diagnosed BPH and by treatment type (surgical, minimally invasive, pharmacological, watchful waiting) assuming all men of the Olmsted County population to be at risk. Temporal changes in incidence were analyzed by Poisson regression. RESULTS: From 1987 to 1997, 2,330 cases of BPH were identified representing an age adjusted, corrected incidence for BPH among male residents of Olmsted County of 854.7 per 100,000 men. While the incidence of BPH increased from 1987 to 1992, coinciding with the introduction of serum prostate specific antigen testing, and subsequently decreased through 1997, the overall incidence of BPH decreased during the 10-year period for the entire cohort (p =0.002). Although the incidence of watchful waiting appeared to increase in the mid 1990s, significant decreases in age adjusted incidence were observed for the period overall (p <0.001). Significant decreases in surgical treatments were observed (p <0.0001). In contrast, there were significant increases in the age adjusted rates of pharmacological treatments during the 10 years (p <0.0001). No significant differences in incidence patterns of treatment type over time were observed by age group. CONCLUSIONS: The introduction of serum prostate specific antigen testing as a screening tool for prostate cancer along with the advent of less invasive alternatives, including pharmacological therapy, have had a significant impact on the incidence of diagnosed BPH in the last decade. PMID- 15879824 TI - Antidiuresis: a new concept in managing female daytime urinary incontinence. PMID- 15879825 TI - Urgency: the cornerstone symptom of overactive bladder. PMID- 15879826 TI - Mixed incontinence. PMID- 15879827 TI - Treatment of urge-predominant mixed urinary incontinence with tolterodine extended release: a randomized, placebo-controlled trial. PMID- 15879829 TI - Impact of urinary incontinence after stroke: results from a prospective population-based stroke register. PMID- 15879831 TI - Andropause: endocrinology, erectile dysfunction, and prostate pathophysiology. PMID- 15879832 TI - The relation of lower urinary tract symptoms with life-style factors and objective measures of benign prostatic enlargement and obstruction: an Italian survey. PMID- 15879833 TI - Are indications for prostatectomy in octogenarians the same as for younger men? PMID- 15879834 TI - Clitoral anatomy in nulliparous, healthy, premenopausal volunteers using unenhanced magnetic resonance imaging. AB - PURPOSE: We determined the magnetic resonance imaging (MRI) characteristics of normal clitoral anatomy. MATERIALS AND METHODS: A series of MRI studies of 10 healthy, nulliparous volunteers with no prior surgery and normal pelvic examination was studied and the key characteristics of clitoral anatomy were determined. A range of different magnetic resonance sequences was used without any contrast agent. RESULTS: The axial plane best revealed the clitoral body and its proximal continuation as the paired crura. The glans was seen more caudal than the body of the clitoris. The bulbs of the clitoris had the same signal as the rest of the clitoris in the axial plane and they related consistently to the other erectile structures. The bulbs, body and crura formed an erectile tissue cluster, namely the clitoris. In turn, the clitoris partially surrounded the urethra and vagina, forming a consistently observed tissue complex. Midline sagittal section revealed the shape of the body, although in this plane the rest of the clitoris was poorly displayed. The coronal plane revealed the relationship between the clitoral body and labia. The axial section cephalad to the clitoral body best revealed the vascular component of the neurovascular bundle to the clitoris. The fat saturation sequence particularly highlighted clitoral anatomy in healthy, premenopausal, nulliparous women. CONCLUSIONS: Normal clitoral anatomy has been clearly demonstrated using noncontrast pelvic MRI. PMID- 15879835 TI - Crural ligation for primary erectile dysfunction: a case series. AB - PURPOSE: Some men with primary erectile dysfunction (ED) have maldevelopment of the crura of the penis. We report our experience with crural ligation for primary venous leakage ED in the last 5 years. MATERIALS AND METHODS: Between December 1998 and March 2004, 11 patients with primary ED underwent crural ligation surgery for congenital venous leakage. All patients were evaluated with pertinent history and physical examinations, color duplex ultrasound, nocturnal penile tumescence study with RigiScan, cavernosometry and cavernosography. Localized crural leakage was the pathognomonic feature in these patients. Surgery involved reflection of the dorsal and cavernous arteries and nerves off of the crura, followed by ligation of the 2 crura proximal to the entrance of the cavernous artery with umbilical tapes. Followup was performed by telephone interview or personal interview at the clinic. A retrospective preoperative and postoperative questionnaire, that is the abridged 5-item version of the 15-item International Index of Erectile Function, was given at the time of the interview. Statistical analysis was performed with the paired t test. RESULTS: Median patient age at surgery was 28 years (range 22 to 39) and mean followup after surgery was 34 months (range 6 to 58). Mean International Index of Erectile Function +/- SD preoperatively and postoperatively was 8.9 +/- 4.5 and 17.5 +/- 5.0, respectively. For questions 2, 3 and 5 mean postoperative scores were significantly better than postoperative scores (p <0.05). Marked improvement in erectile function was noted in 9 of our 11 patients. CONCLUSIONS: Penile venous surgery may not have a lasting result in men in whom venous leakage is due to systemic diseases or penile smooth muscle atrophy. However, in patients with a congenital venous leak abnormality of the crura ligation of the proximal crura with umbilical tapes seems to have a satisfactory, durable result. PMID- 15879836 TI - Vardenafil improved patient satisfaction with erectile hardness, orgasmic function and sexual experience in men with erectile dysfunction following nerve sparing radical prostatectomy. AB - PURPOSE: Nerve sparing radical retropubic prostatectomy (NS-RRP) results in erectile dysfunction in a significant number of patients. Vardenafil, a potent and selective phosphodiesterase type 5 inhibitor, is generally safe. It improves International Index of Erectile Function erectile function domain scores, and penetration and erection maintenance success rates in patients who have undergone NS-RRP. We report additional parameters important to patient perceptions regarding erection quality and satisfaction with sexual experience following NS RRP. MATERIALS AND METHODS: A total of 440 men at 58 centers throughout the United States and Canada participated in this randomized, placebo controlled, double-blind trial with 3 phases, namely baseline (4-week untreated period), treatment (12 weeks) and followup (7 days). Participants received placebo (145), 10 mg vardenafil (146) or 20 mg vardenafil (149) at home on demand but no more than once per calendar day. Efficacy and satisfaction with erection quality and sexual experience were determined during the trial. RESULTS: The 10 and 20 mg vardenafil doses were significantly superior to placebo for the International Index of Erectile Function domains for intercourse satisfaction, orgasmic function and overall satisfaction with sexual experience (vs placebo p <0.0009). Significant improvement in the satisfaction rate with erection hardness were demonstrated for each vardenafil dose compared with placebo (p <0.0001). Vardenafil was generally well tolerated. Common adverse events were headache, vasodilatation and rhinitis. CONCLUSIONS: In this difficult to treat population of men with erectile dysfunction subsequent to NS-RRP on demand treatment with vardenafil during a 3-month period significantly improved key aspects of the sexual experience important to patient quality of life. PMID- 15879837 TI - Outcome of gonadotropin therapy for male hypogonadotropic hypogonadism at university affiliated male infertility centers: a 30-year retrospective study. AB - PURPOSE: Hypogonadotropic hypogonadism (HH) is an uncommon cause of male infertility. Even specialized centers see small numbers of patients and have little experience in restoring fertility in these patients. To explore the phenotypic heterogeneity and treatment outcome of HH, we summarized our 30-year experience. MATERIALS AND METHODS: Medical charts of 36 male patients (age range 11 to 42 years) with HH (81% primary, 19% secondary) treated at university affiliated male infertility centers were reviewed. Pubic hair stage, genital stage, testicular volume (TV), sexual function and sperm production were determined before and after human chorionic gonadotropin/human menopausal gonadotropin (hCG/hMG) treatment. Differences were analyzed statistically. RESULTS: The population was stratified according to initial TV into a small testis subset (TV less than 4 ml in 23) and a large testis subset (TV 4 ml or greater in 13). Comparison of these 2 subsets revealed significant differences in the incidence of cryptorchidism (39% vs 8%) and Kallmann's syndrome (22% vs 0%), pubic hair stage (1.3 +/- 0.5 vs 3.5 +/- 1.4), genital stage (1.2 +/- 0.4 vs 3.9 +/- 0.9) and TV (1.6 +/- 0.9 vs 7.5 +/- 3.5 ml). Long-term administration of hCG/hMG for 12 to 240 months (average 56 +/- 11) resulted in sperm production in only 36% of the small testis subjects but in 71% of the large testis subjects. CONCLUSIONS: Initial TV values provide insight into phenotypic variables such as time of onset and severity in patients with primary or secondary HH, and may predict sperm output in response to hCG/hMG therapy. PMID- 15879838 TI - Incidentally diagnosed Peyronie's disease in men presenting with erectile dysfunction. PMID- 15879839 TI - Erectile dysfunction in smokers: a penile dynamic and vascular study. PMID- 15879840 TI - Association between lower urinary tract symptoms and erectile dysfunction. PMID- 15879841 TI - Effects of glans penis augmentation using hyaluronic acid gel for premature ejaculation. PMID- 15879843 TI - Psycho-biological correlates of rapid ejaculation in patients attending an andrologic unit for sexual dysfunctions. PMID- 15879844 TI - Microsurgical treatment of persistent or recurrent varicocele. PMID- 15879847 TI - A clinical pathway for laparoscopic pyeloplasty decreases length of stay. AB - PURPOSE: Benefits of minimally invasive procedures include decreased hospitalization and recovery times. Decreased length of stay (LOS) improves hospital efficiency and decreases costs. However, decreasing the LOS at the expense of patient care and satisfaction is not acceptable. A clinical pathway (CP) with structured order sets and imaging was developed for patients undergoing laparoscopic pyeloplasty. This pathway includes a cascade of activities managed closely by the health care team. This study assesses the safety and patient satisfaction with this clinical pathway. MATERIALS AND METHODS: We reviewed all adult pyeloplasties (39) completed laparoscopically since November 2001. All patients were managed according to the CP developed for the laparoscopic pyeloplasty procedure. The length of stay was measured in days. Patient satisfaction was assessed with a standardized questionnaire. Any readmissions or emergency room visits were documented. RESULTS: The mean length of stay was 1.10 days. Of 39 patients 37 (94%) were discharged home on postoperative day 1. One patient with severe postoperative pain required intravenous analgesia. She had undergone complex upper tract reconstruction and stayed a total of 4 days. One patient, who had a previous failed endopyelotomy, remained 2 days for persistent nausea. No patients sought emergency room consultation and there were no readmissions. Of 39 patients 34 (87%) completed the questionnaire and satisfaction was high. CONCLUSIONS: The implementation of a CP at our institution has standardized patient care in this population and decreased LOS in comparison to the literature. This improves bed use and hospital efficiency while maintaining a high degree of patient satisfaction. We conclude that with intensive patient care and education most patients undergoing laparoscopic pyeloplasty may be discharged home safely on postoperative day 1. PMID- 15879848 TI - Impact of malpractice caps on use and outcomes of radical cystectomy for bladder cancer: data from the surveillance, epidemiology, and end results program. AB - PURPOSE: The impact of malpractice awards on insurance premiums and health care delivery generates much concern. To our knowledge no data exist regarding the impact of caps, or upper limits, on awards for noneconomic damages (also termed pain and suffering) on health care delivery patterns or outcomes. We investigated the effect of caps on the use of and outcomes following aggressive surgical treatment (radical cystectomy) in patients with bladder cancer. MATERIALS AND METHODS: We performed a retrospective cohort study of patients with bladder cancer who underwent radical cystectomy, identified from the Surveillance, Epidemiology, and End Results (SEER) Program database. Cystectomy rates and post cystectomy disease specific survival were compared between SEER regions with and without a cap, while controlling for other variables. RESULTS: A significantly greater proportion of patients with stages III and IV bladder cancer underwent cystectomy in SEER regions with a cap. Cap status was a significant predictor of survival from bladder cancer. CONCLUSIONS: Radical cystectomy for bladder cancer is performed more often even for advanced bladder cancer in geographic regions with a cap with a positive impact on survival. The institution of caps may have beneficial effects on patterns of health care beyond that of merely decreasing insurance premium costs. PMID- 15879849 TI - A cost comparison of medical management and transurethral needle ablation for treatment of benign prostatic hyperplasia during a 5-year period. AB - PURPOSE: We performed an analysis comparing the cost of medical management with TUNA therapy for a 5-year period. MATERIALS AND METHODS: Published costs for tamsulosin, finasteride, transurethral needle ablation (TUNA, Medtronic, Inc., Minneapolis, Minnesota) and transurethral resection of the prostate were used to construct a cost analysis model comparing medication with TUNA. The model analyzed monotherapy with an alpha-blocker (tamsulosin) and a 5alpha-reductase inhibitor (finasteride), combination therapy using both medications, and a mixed scenario using monotherapy and combination therapy. Published data were used to estimate the rate of surgical intervention in patients initially treated with medications or TUNA. RESULTS: Tamsulosin monotherapy was less expensive than TUNA for 5 years ($3,485 for tamsulosin vs $4,811 for TUNA year 5). Finasteride monotherapy reaches a break-even point with TUNA during year 5 ($4,867 for finasteride vs $4,811 for TUNA). Combination therapy reaches a break-even point with TUNA after approximately 2 years 7 months of treatment ($4,515 for combination therapy vs $4,572 for TUNA) and the mixed scenario breaks even with TUNA at approximately year 4 ($4,696 for medical management vs $4,645 for TUNA). CONCLUSIONS: The TUNA procedure compares favorably to combination medical therapy for the treatment of benign prostatic hyperplasia on a cost basis. alpha-Blocker monotherapy is less costly than TUNA while 5alpha-reductase inhibitor monotherapy is approximately equivalent to TUNA for 5 years. The TUNA procedure is less expensive than combination medical management for 5 years, with a break-even point at approximately 2 years 7 months. PMID- 15879850 TI - The effect of hospital volume on cancer control after radical prostatectomy. AB - PURPOSE: For complex oncological procedures, hospital volume affects short and long-term patient outcome. We examined the association of hospital volume and long-term cancer control after radical prostatectomy. MATERIALS AND METHODS: With a cohort study design, we used the Surveillance, Epidemiology and End Results Medicare linked files to identify a population based sample of men with newly diagnosed prostate cancer treated primarily with radical prostatectomy. Failure of cancer control was defined as the use of postoperative medical or surgical hormone ablation or treatment with radiation therapy more than 6 months after surgery. RESULTS: A total of 12,635 men underwent radical prostatectomy for incident prostate cancer. After adjusting for age, comorbidity, histological grade and clinical stage, the risk of adjuvant therapy was greater among those treated at low (1 to 33 cases) and medium (34 to 61 cases) volume hospitals than at very high (more than 108 cases) volume hospitals (HR 1.25, p <0.001 and HR 1.11, p =0.023 respectively). CONCLUSIONS: Patients treated at lower volume institutions are at increased risk of initiation of subsequent adjuvant therapy with radiation therapy, medical hormone ablation or orchiectomy. Noted differences in cancer control provide additional evidence regarding issues surrounding the debate over surgical volume standards for the surgical treatment of prostate cancer. PMID- 15879851 TI - Variations among high volume surgeons in the rate of complications after radical prostatectomy: further evidence that technique matters. AB - PURPOSE: A strong association between surgeon, hospital volume and postoperative morbidity of radical prostatectomy has been demonstrated. While better outcomes are associated with high volume surgeons, the degree of variation in outcomes among surgeons has not been fully examined. MATERIALS AND METHODS: Using a linked database from Surveillance, Epidemiology and End Results registries and federal Medicare claims data, we analyzed outcomes of consecutive patients treated with radical prostatectomy between 1992 and 1996. We focused on variations in several measures of morbidity (perioperative complications, late urinary complications and long-term incontinence) among patients of high volume surgeons, defined as those with 20 or more patients in the study period. After adjusting for hospital, surgeon volume and case mix, we examined the extent to which variations in the rates of adverse outcomes differed among surgeons for all 3 end points. RESULTS: Of the 999 surgeons 16% (159) performed 48.7% (5,238) of the 10,737 radical prostatectomies during the study. The 30-day mortality rate was 0.5%, the major postoperative complication rate was 28.6%, late urinary complications 25.2% (major events 16%) and long-term incontinence 6.7%. For all 3 morbidity outcomes the variation among surgeons in the rate of complications was significantly greater than that expected by chance (p =0.001 for each) after adjustment of covariates. Furthermore, surgeons with better (or worse) than average results with regard to 1 outcome were likely to have better (or worse, respectively) results with regard to the other 2 outcome measures. CONCLUSIONS: Morbidity end points that directly affect quality of life showed significant variability among high volume providers. Surgeons who performed well in 1 area (eg postoperative complications) performed well in others. These results further suggest that variations in surgical technique and postoperative care lead to variations in outcomes after radical prostatectomy, indicating that outcomes of this operation are sensitive to small differences in performance. PMID- 15879852 TI - Healthcare use and costs of primary and secondary care patients with prostatitis. PMID- 15879857 TI - Early cyclosporine a withdrawal in kidney-transplant recipients receiving sirolimus prevents progression of chronic pathologic allograft lesions. PMID- 15879858 TI - The natural history of chronic allograft nephropathy. PMID- 15879860 TI - Increasing the ranks of physician-scientists in urology through the promotion of fellowship training: the example of pediatric urology. PMID- 15879861 TI - An artificial somatic-autonomic reflex pathway procedure for bladder control in children with spina bifida. AB - PURPOSE: Neurogenic bladder is a major problem for children with spina bifida. Despite rigorous pharmacological and surgical treatment, incontinence, urinary tract infections and upper tract deterioration remain problematic. We have previously demonstrated the ability to establish surgically a skin-central nervous system-bladder reflex pathway in patients with spinal cord injury with restoration of bladder storage and emptying. We report our experience with this procedure in 20 children with spina bifida. MATERIALS AND METHODS: All children with spina bifida and neurogenic bladder underwent limited laminectomy and a lumbar ventral root (VR) to S3 VR microanastomosis. The L5 dorsal root was left intact as the afferent branch of the somatic-autonomic reflex pathway after axonal regeneration. All patients underwent urodynamic evaluation before and after surgery. RESULTS: Preoperative urodynamic studies revealed 2 types of bladder dysfunction- areflexic bladder (14 patients) and hyperreflexic bladder with detrusor external sphincter dyssynergia (6). All children were incontinent. Of the 20 patients 17 gained satisfactory bladder control and continence within 8 to 12 months after VR microanastomosis. Of the 14 patients with areflexic bladder 12 (86%) showed improvement. In these cases bladder capacity increased from 117.28 to 208.71 ml, and mean maximum detrusor pressure increased from 18.35 to 32.57 cm H2O. Five of the 6 patients with hyperreflexic bladder demonstrated improvement, with resolution of incontinence. Urodynamic studies in these cases revealed a change from detrusor hyperreflexia with detrusor external sphincter dyssynergia and high detrusor pressure to nearly normal storage and synergic voiding. In these cases mean bladder capacity increased from 94.33 to 177.83 ml, and post-void residual urine decreased from 70.17 to 23.67 ml. Overall, 3 patients failed to exhibit any improvement. CONCLUSIONS: The artificial somatic autonomic reflex arc procedure is an effective and safe treatment to restore bladder continence and reverse bladder dysfunction for patients with spina bifida. PMID- 15879862 TI - Simplified mons plasty: a new technique to improve cosmesis in females with the exstrophy-epispadias complex. AB - PURPOSE: Long-term data suggest that a significant number of women with the exstrophy-epispadias complex (EEC) report dissatisfaction with the cosmetic appearance of the genitalia. We developed a new technique of mons plasty in an attempt to improve the overall cosmetic appearance of the external genitalia in females with EEC. MATERIALS AND METHODS: Nine consecutive females with a mean age of 6.4 years (range 2 days to 12.2 years) previously diagnosed with EEC underwent simplified mons plasty concomitantly with various other urological reconstructive procedures. Briefly, a simplified mons plasty was performed by approximating the bifid hemiclitoris and subsequently incising the lateral sulci between the labia majora and minora, which allowed the superomedial rotation of the labia majora and underlying peripubic adipose tissue, thus, recreating the mons. RESULTS: There were no perioperative complications. With a mean followup of 6 months (range 4 to 12) all patients had improved cosmesis with respect to a more prominent mons, more appropriately situated labia and adequately recessed introitus. CONCLUSIONS: Simplified mons plasty improves cosmesis in females with external genital abnormalities secondary to EEC. Although our followup is admittedly short, we believe that this technique should be considered in the armamentarium of the reconstructive surgeon during the operative treatment of this complex group of patients. PMID- 15879863 TI - Double-blind placebo controlled study of alpha-adrenergic receptor antagonists (doxazosin) for treatment of voiding dysfunction in the pediatric population. AB - PURPOSE: We sought to determine whether the alpha-adrenergic antagonist doxazosin could be used as primary therapy in children with voiding dysfunction. MATERIALS AND METHODS: Children were assigned to maintain a voiding diary and then randomly divided into a double-blind placebo controlled protocol (0.5 mg doxazosin or placebo). Duplicate uroflow studies with post-void residual evaluations and assessment of dysfunctional voiding scores were performed on initiation and completion of the study. At the conclusion parents were asked to rank the perceived improvement of the urinary incontinence (ie parental subjective perception of improvement). RESULTS: No significant differences between doxazosin (18) and placebo (20) treated patients were found in the number of incontinent days per week, severity of incontinent episodes or alterations in uroflow patterns. Although not significant, 2 findings suggested a beneficial effect of doxazosin over placebo. Specifically, doxazosin decreased the number of incontinent episodes weekly from a median of 18 to 4, while the number of incontinent episodes weekly in the placebo group remained essentially unchanged, decreasing from 15 to 14 (p = 0.13). Doxazosin also improved the dysfunctional voiding scores over placebo, for an improvement of -3 vs 0 points. Further substantiating a doxazosin effect over placebo was the subjective perception of the parents that doxazosin significantly improved urinary continence (p <0.02). CONCLUSIONS: Compared to placebo, doxazosin did not demonstrate a significant objective benefit, but produced a significant subjective benefit in the treatment of urinary incontinence secondary to voiding dysfunction. PMID- 15879864 TI - The effect of oxybutynin treatment on cognition in children with diurnal incontinence. AB - PURPOSE: Oxybutynin is a powerful anticholinergic drug already known to impair cognition in the elderly. The impact of this drug on cognitive functioning in the pediatric population is unknown. We report the results of a study designed to assess the effect of oxybutynin on cognitive function in children. MATERIALS AND METHODS: A total of 25 patients presenting with the primary symptom of daytime enuresis were recruited for this nonrandomized trial. All subjects initially received 4 weeks of behavior modification, followed by an additional 4 weeks of behavior modification either alone or with oxybutynin for continued treatment of enuresis. Neuropsychological testing was performed at baseline (4 weeks) and after additional therapy (8 weeks). RESULTS: Patient demographics included a male to-female ratio of 11:14 and a mean age of 7.2 +/- 1.8 years. A total of 10 patients were assigned to the control group receiving behavior modification, and 15 patients were assigned to the treatment group receiving behavior modification plus oxybutynin. The oxybutynin treated patients had a lower overall performance at baseline pretreatment testing. However, performance in this group improved following treatment with oxybutynin. CONCLUSIONS: Oxybutynin, a commonly used pharmacological agent in pediatric urology, was not associated with cognitive impairment following treatment. However, we observed lower baseline cognitive functioning in patients whose parents chose oxybutynin over behavior modification alone. This finding may represent a selection bias. However, it also supports the need for a multidisciplinary approach to the treatment of patients with dysfunctional voiding, as some may have cognitive difficulties that have not previously been explored. PMID- 15879865 TI - Bladder neck sling for treatment of neurogenic incontinence in children with augmentation cystoplasty: long-term followup. AB - PURPOSE: We retrospectively reviewed the outcome and long-term followup (mean 4.16 years) of bladder neck slings for the treatment of neurogenic urinary incontinence in 58 patients (15 males) who also underwent bladder augmentation. MATERIALS AND METHODS: A total of 58 patients with neurogenic bladder (43 females and 15 males, median age 11.4 years) underwent a rectus fascial sling procedure as part of the reconstructive efforts for continence between July 1991 and July 2003. Criteria for enhancement of bladder outlet resistance included a detrusor leak point pressure of less than 45 cm H2O, an open bladder neck during bladder filling at low detrusor pressures and clinical evidence of stress incontinence. RESULTS: Followup ranged from 1 year to 10 years, 3 months (mean 4.16 years). A total of 51 patients (88%) obtained good continence results. Five females and 2 males remained incontinent following the sling procedure. Four females underwent a secondary open bladder neck procedure at a mean of 18 months after the initial procedure (artificial urinary sphincter in 2, bladder neck closure in 2). Two male patients (5 and 17 years old) had daily underwear staining or dampness with exercise or transfer. CONCLUSIONS: We consider bladder neck slings the procedure of choice for the enhancement of bladder outlet resistance in the majority of patients with neurogenic bladder who need augmentation cystoplasty and whom we do not expect will be capable of voiding spontaneously. In males and females satisfactory long-term continence can be expected with the use of the rectus fascial sling. PMID- 15879866 TI - Dysfunctional elimination syndrome as an etiology of idiopathic urethritis in childhood. AB - PURPOSE: Idiopathic urethritis (IU) of childhood or urethrorrhagia is a common problem characterized by blood spotting in the underwear between voiding. A clear etiology has not been established and treatments vary. We postulate that idiopathic urethritis is a manifestation of underlying dysfunctional elimination syndrome (DES). MATERIALS AND METHODS: During a 5-year period we reviewed the records of all children diagnosed with IU in our practice. In total 72 children fit the analysis criteria. There were 68 boys and 4 girls. All children presented with either gross blood per urethra or microhematuria. Children with active infection, immunodeficiency, neurogenic bladder, vesicoureteral reflux, infravesical obstruction, urethral trauma or other genitourinary anomalies were excluded. Evaluation included thorough history and physical examination, urinalysis and urine culture. Renal and bladder ultrasound, voiding cystourethrogram and uroflow/electromyogram/post-void residual volume were obtained in select patients. Study children were divided into 2 cohorts. The first cohort (group 1, 37 patients) was treated with traditional remedies using antibiotics, urinary analgesics and/or anticholinergics. The second cohort (group 2, 35 patients) was treated by bowel and bladder regimens, laxatives when necessary, and biofeedback and/or alpha-blockers when sphincter dyssynergia was identified. RESULTS: A total of 13 patients in group 1 (35%) had a full response to treatment, 6 (16%) had a partial response and 18 (49%) failed to respond. A total of 29 patients in group 2 (83%) had a full response to treatment, 2 (6%) had a partial response and 4 (11%) had no response. It took an average of 12.1 months to respond fully in group 1, while in group 2 the same full response took an average of 5.2 months. Of the 18 children who crossed over from group 1 to group 2, 15 (83%) had a full response with an average response time of 7.3 months. CONCLUSIONS: Our data clearly reveal a higher cure rate when children with urethritis are treated according to DES guidelines. IU of childhood is a manifestation of underlying DES and should be treated as such. PMID- 15879867 TI - Clinical features and immunoexpression of p53, MIB-1 and proliferating cell nuclear antigen in adrenal neoplasms. AB - PURPOSE: We evaluated the clinical features and immunoreactivity of p53 protein, MIB-1 antigen and proliferating cell nuclear antigen (PCNA) in adrenal neoplasms. MATERIALS AND METHODS: A total of 26 patients with adrenocortical adenoma and 24 patients with carcinoma were treated with adrenalectomy. Clinical features and immunohistochemical reactions were compared in adult vs pediatric tumors. RESULTS: There was a bimodal age distribution of carcinomas and adenomas, with a first peak occurring before age 5 years. The proportion of carcinomas in children (18 of 29) was higher than in adults (6 of 21). Carcinoma and adenoma occurring in children presented more commonly as the virilizing syndrome, while in adults Cushing's syndrome was more common. All adenomas in adults were p53 negative, while in children 4 of 11 adenomas (36%) were p53 positive. Histological Weiss criteria were the most reliable pathological features to distinguish adenoma from carcinoma. Other pathological features, including tumor weight, rate of mitotic figures and immunoexpression of p53 protein, MIB-1 antigen and PCNA, exhibited a striking difference in adenomas and carcinomas but none demonstrated sensitivity or specificity of 100%. Of all the computerized tomographic characteristics analyzed, including tumor size, shape, necrosis/hemorrhage, attenuation and contrast enhancement, only tumor size (greater than 5 cm) showed sensitivity and specificity of 100% in the differential diagnosis. Children and adults with carcinoma had similar curves of survival (p = 0.76). Carcinoma stage and PCNA immunoexpression displayed an association with outcome. CONCLUSIONS: Endocrine syndromes differed in adults and children but other clinical features were similar in both groups. The role of p53 protein, MIB-1 antigen and proliferating cell nuclear antigen in discrimination of adenomas from carcinomas is unclear. PMID- 15879868 TI - Outcome of valve ablation in late-presenting posterior urethral valves. PMID- 15879869 TI - Modification of meatal advancement and glanuloplasty for correction of distal hypospadias. PMID- 15879871 TI - Pelvic fracture-associated urethral injuries in girls: experience with primary repair. PMID- 15879872 TI - Vaginal construction using sigmoid colon in children and young adults. PMID- 15879873 TI - Oral complications after buccal mucosal graft harvest for urethroplasty. PMID- 15879875 TI - Wilms tumor. PMID- 15879876 TI - Computerized tomography diagnosis of right ureteral calculus and coexisting acute appendicitis. PMID- 15879877 TI - CD70: a new tumor specific biomarker for renal cell carcinoma. AB - PURPOSE: To date there have been no specific tumor markers available for the differential diagnosis of renal cell carcinoma (RCC). In an earlier study we identified high RNA expression of CD70 in clear cell RCC. CD70 is a type II transmembrane protein belonging to the tumor necrosis factor family. It represents the ligand for CD27, a glycosylated transmembrane protein of the tumor necrosis factor receptor family. To our knowledge the function of CD70 in solid tumors is not known. In the current study we analyzed CD70 protein expression in different RCC subtypes. MATERIALS AND METHODS: A total of 68 tumor samples of different histopathological subtypes were investigated by immunochemistry, including 41 clear cell, 19 papillary and 5 chromophobe RCCs, and 3 oncocytomas as well as their normal tissue counterparts. Immunochemistry was performed on frozen tissue samples using monoclonal antibody against CD70. RESULTS: None of the normal kidney tissues showed CD70 expression. In contrast, all clear cell RCCs expressed CD70 at a high level. Positive immunostaining was observed in 1 papillary (5%) and in 1 chromophobe (20%) RCC. Five papillary tumor samples (26%) showed focal staining in less than 5% of cells. All other samples were negative for CD70. CONCLUSIONS: Our study identified CD70 as a new specific tumor marker for clear cell RCC. This new marker can be used for differential diagnosis in cases of uncertain histological classification. The function of this protein in tumorigenesis and its use as a diagnostic marker in serum and urine or as a therapeutic tool must be investigated in further studies. PMID- 15879878 TI - Expression of gastrin releasing Peptide receptor in renal cell carcinomas: a potential function for the regulation of neoangiogenesis and microvascular perfusion. AB - PURPOSE: Gastrin releasing peptide (GRP) is a growth factor for renal cell carcinoma (RCC) and it has vasoactive properties. Blockade of GRP receptor inhibits the growth of GRP receptor positive and negative tumors in nude mice, suggesting GRP effects other than those related to tumor epithelium. Therefore, in this study we analyzed the effects of GRP receptor blockade on neoangiogenesis in RCC. MATERIALS AND METHODS: GRP receptor expression was determined in human RCC and corresponding normal tissue by real-time reverse transcriptase-polymerase chain reaction, immunohistochemistry and confocal laser scanning microscopy. Multicellular spheroids of the A498 RCC line were implanted into dorsal skin fold chambers of athymic nude mice. Neoangiogenesis was measured by intravital microscopy after blockade of GRP receptors by the GRP antagonist RC-3095. The influence of GRP on vascular endothelial growth factor secretion in A498 cells was studied in vitro. RESULTS: GRP receptor expression was immunolocalized in tumor cells and microvessels. Implanted tumor cell spheroids and spheroid microvessels of the chamber also expressed GRP receptors. Spheroid neoangiogenesis was significantly inhibited by RC-3095 when given immediately after spheroid implantation. Vascular endothelial growth factor secretion of A498 cells was not affected by GRP. CONCLUSIONS: RCC angiogenesis is sensitive to GRP receptor blockade. Therefore, GRP receptors may not only stimulate tumor cell proliferation, but also affect tumor microcirculation. PMID- 15879879 TI - Renal cryoablation and radio frequency ablation: an evaluation of worst case scenarios in a porcine model. AB - PURPOSE: Although ablative technologies, including radio frequency (RF) ablation (RFA) and cryoablation (CA), are being used to treat renal masses, complications associated with injury to vital renal structures are not well understood. We investigated these worst case scenarios by deliberately targeting vital renal structures with CA or RFA in a porcine model. MATERIALS AND METHODS: Following surgical exposure of the right kidney in female pigs a cryoneedle or an RF probe was deliberately placed under visual and ultrasound guidance in the renal pelvis (CA in 5 pigs and RFA in 7), major calix (CA and RFA in 5 each) or subsegmental renal vessels (CA in 5 pigs and RFA in 7). Cryo-energy or RF energy was then applied to create a 3 cm lesion. After 10 days the kidneys underwent gross and histological examination for urine and blood extravasation, cell death and injury. Ex vivo retrograde pyelography was performed to evaluate for urinary fistulas. RESULTS: All pigs tolerated the treatment and no procedure related deaths occurred. No significant bleeding was noted. RFA and CA created reproducible lesions and areas of cell death and necrosis. Despite significant intentional injury to the collecting system no urinary fistulas were demonstrated in CA specimens (0 of 15). In contrast, damage to the renal pelvis (4 of 7) by dry (3 of 4) or wet (1 of 3) RFA was associated with a high likelihood of urinary extravasation. CONCLUSIONS: This short-term study demonstrates that CA is safe, effective and not associated with urinary extravasation. In contrast, RFA to the renal pelvis is associated with urinary extravasation. Further studies are needed to support these findings. PMID- 15879880 TI - Mitomycin C and epirubicin: functional bladder damage in rats after repeat intravesical instillations. AB - PURPOSE: We studied functional damage in the bladder after intravesical administration of mitomycin C or epirubicin in rats. MATERIALS AND METHODS: A total of 40 rats were divided into 4 groups. Rats in 3 groups received intravesical instillations of mitomycin C, epirubicin or saline. Rats in group 4 received no intravesical instillations and served as normal controls. The doses of drugs administered were based on those used for therapeutic purposes. Changes in vesical capacity and bladder wall compliance were determined. Compliance was defined as the change in vesical volume induced by a given change in pressure. RESULTS: Weekly instillations of 1 mg/ml mitomycin C caused a statistically significant decrease in vesical volume and compliance, which persisted 3 weeks after the cessation of therapy. Epirubicin also induced changes in vesical volume and compliance but only that in bladder compliance achieved statistical significance. These changes were not seen in NaCl treated animals or in normal controls. CONCLUSIONS: Repeat intravesical instillations of mitomycin C have a prolonged effect on bladder function that is not seen after saline solution or epirubicin instillation. PMID- 15879881 TI - Activation of natural killer T cells by alpha-galactosylceramide mediates clearance of bacteria in murine urinary tract infection. AB - PURPOSE: alpha-Galactosylceramide (alpha-GalCer), which is a specific ligand for CD1d restricted variable-alpha14chain natural killer T cells, has an important role in host defense against a range of microbial infections. We examined whether alpha-GalCer mediates bacterial clearance in a murine urinary tract infection (UTI) model. MATERIALS AND METHODS: The murine UTI model was established by intravesical inoculation of Escherichia coli, Pseudomonas aeruginosa or methicillin resistant Staphylococcus aureus, followed by clamping the distal end of the urethra of C57BL/6 female mice for 4 hours. The antibacterial effect of alpha-GalCer was assessed by comparing the number of cfu/gm kidney tissue 4 days after bacterial inoculation. The prophylactic effect of alpha-GalCer was examined by administrating 3 doses of intraperitoneal alpha-GalCer on alternate days 24 hours before E. coli inoculation. The therapeutic effect of alpha-GalCer was tested by the administration of 2 doses of intraperitoneal alpha-GalCer after bacterial inoculation. To assess cytokine induction by alpha-GalCer serum levels of interleukin-12, interferon-gamma and tumor necrosis factor-alpha were measured by cytometric bead array assay. RESULTS: When administered before bacterial inoculation, alpha-GalCer showed a strong prophylactic antibacterial effect. alpha-GalCer also showed a marked antibacterial effect on preestablished mice UTI caused by E. coli, P. aeruginosa and methicillin resistant S. aureus. alpha GalCer induced a significantly higher level of interleukin-12, interferon-gamma and tumor necrosis factor-alpha compared with control glycolipids. CONCLUSIONS: These findings suggest a significant role for alpha-GalCer in regulating antibacterial functions by activating natural killer T cells in the murine UTI model. PMID- 15879882 TI - Can the Bonn Risk Index be replaced by a simple measurement of the urinary concentration of free calcium ions? AB - PURPOSE: It has been shown that a strong hyperbolic relationship exists between the urinary concentration of free Ca ions ([Ca]) and the amount of ammonium oxalate (Ox) that must be titrated in a standardized procedure to a urine sample to induce CaOx crystallization. The ratio of [Ca] to (Ox) is termed the Bonn Risk Index (BRI). Most data plot around a hyperbola described by the formula, [Ca] x (Ox) = constant. Due to the high relationship between [Ca] and (Ox) one may argue that determining only 1 of the 2 BRI parameters, preferably [Ca], is sufficient to describe the urine crystallization risk. MATERIALS AND METHODS: Based on 195 urine samples taken from CaOx stone formers and healthy subjects we compared the sensitivity and specificity of BRI, and its corresponding [Ca] value by calculating ROC curves. Furthermore, ROC curves of the established risk indexes, namely the model value of urinary supersaturation and the model value of the urine activity product, are presented. RESULTS: Our results clearly demonstrate that 1) BRI has the highest sensitivity and specificity of the tested indexes, 2) (Ox) cannot be reliably predicted from [Ca] and 3) determining [Ca] alone revealed a meaningful first estimate of urine CaOx crystallization risk according to BRI. CONCLUSIONS: To avail ourselves of the high quality of BRI in patient treatment the additional determination of (Ox) is required. PMID- 15879883 TI - A role for muscarinic receptors or rho-kinase in hypertension associated rat bladder dysfunction? AB - PURPOSE: Essential arterial hypertension is a frequent condition. Spontaneously hypertensive rats (SHRs) show bladder dysfunction similar to that seen in patients with overactive bladder. Since muscarinic receptors and rho-kinase have a key role in the regulation of bladder contractility, we determined whether alterations of either one might contribute to hypertension associated bladder dysfunction. MATERIALS AND METHODS: The bladders of SHRs and normotensive Wistar Kyoto rats (WKYs) were compared in in vitro radioligand binding and contractility studies. RESULTS: The mean total number of muscarinic receptors +/- SEM (181 +/- 14 vs 191 +/- 22 fmol/mg protein) and the relative roles of their subtypes were similar in SHRs and WKYs. Contractile responses to the muscarinic agonist carbachol (maximum effect 2.04 +/- 0.24 vs 2.05 +/- 0.14 mN/mm strip length and log EC50 5.61 +/- 0.07 vs 5.64 +/- 0.04) and to KCl in a receptor independent manner were similar in the 2 strains. The M3 selective antagonist darifenacin inhibited carbachol responses much more potently than the M2 selective antagonist methoctramine but the potency of the 2 drugs was similar in each strain. The rho kinase inhibitor Y27,632 attenuated carbachol induced contraction in a quantitatively similar manner in SHRs and WKYs. CONCLUSIONS: An altered function of muscarinic receptor subtypes or rho-kinase does not appear to contribute to bladder dysfunction in SHRs. PMID- 15879884 TI - Cholinergic and purinergic responses in isolated human detrusor in relation to age. AB - PURPOSE: We investigated whether the contractility of isolated human detrusor muscle, responsiveness to commonly used spasmolytic drugs, and expression of selected muscarinic and purinergic (P2X) receptor subtypes (M2, M3, P2X1 and P2X3) change with age. MATERIALS AND METHODS: Tissues were taken from 63 patients 37 to 84 years old undergoing radical cystectomy. Specimens from 49 patients were used for contractility studies and those from 50 were used for mRNA analysis. RESULTS: Propiverine, oxybutynin, tolterodine and atropine decreased contractions evoked by electric field stimulation to different degrees. However, neither the efficacy nor potency of the drugs showed age related changes. Since human detrusor muscle shows atropine resistant noncholinergic responses, we also studied the putative age dependence of concentration-response curves to the muscarinic agonist carbachol, and the purinergic agonists adenosine triphosphate (ATP) and alpha-beta-methylene-ATP. Sensitivity to alpha-beta-methylene-ATP increased with age, while the efficacy and potency of spasmolytic drugs did not depend on age. In addition, mRNA detected for M2, M3, P2X1 and P2X3 receptors did not change with age. CONCLUSIONS: Our results do not provide evidence for age related contractile deterioration in human detrusor muscle strips, nor do they suggest that responses to anticholinergic spasmolytic drugs change substantially with age. PMID- 15879885 TI - Expression of androgen receptor associated protein 55 (ARA55) in the developing human fetal prostate. AB - PURPOSE: Development and differentiation of the human fetal prostate are androgen dependent and follow a specific pattern of solid bud-ductal morphogenesis, which involves stromal-epithelial interactions. Androgen receptor associated protein 55 (ARA55) an androgen receptor coactivator localized in stromal cells, binds to androgen receptor (AR) and regulates androgen receptor translocation and transcriptional activity. We investigated whether ARA55 has a role in human prostate development. MATERIALS AND METHODS: ARA55 expression was examined in 25 human prostates from fetuses at gestational ages 10 to 40 weeks and compared to the expression of 34betaE12 (a basal cell marker), smooth muscle actin, desmin (a smooth muscle marker), vimentin (a mesenchymal marker) and Ki-67 (a proliferation marker) by immunohistochemistry. RESULTS: Prostatic epithelium appeared as solid epithelial buds from the urogenital sinus. It underwent arborization and ductal differentiation from the center to the periphery. ARA55 was expressed in stromal cells with a zonal pattern, primarily in the peripheral zone surrounding the noncanalized acini. Most cells in solid buds were positive for 34betaE12, while only basal layer cells in the centrally located epithelial ducts stained with 34betaE12. Solid buds also had a higher proliferation index than ducts. In addition, ARA55 expressing stromal cells but not ARA55 negative stromal cells showed smooth muscle differentiation. CONCLUSIONS: The intimate relationship between ARA55 expressing stromal cells and mitotically active, noncanalized acini suggests that ARA55 has a role in the stromal-epithelial interaction involved in fetal prostate development. PMID- 15879886 TI - Effects of the antidepressant St. John's wort (Hypericum perforatum) on rat and human vas deferens contractility. AB - PURPOSE: Since sexual dysfunction related to vas deferens smooth muscle contractility is a possible side effect of St. John's wort (SJW) (Hypericum perforatum) we evaluated the effect of this herbal antidepressant on rat and human vas deferens contractility. MATERIALS AND METHODS: The effect of SJW was evaluated on contractions induced by electrical field stimulation or exogenous agonists (alpha,beta-methylene adenosine triphosphate and phenylephrine) in isolated rat and human vas deferens. RESULTS: SJW (1 to 300 microM) decreased in a concentration dependent manner the amplitude of electrical field stimulation and agonist induced contractions with the same potency, suggesting direct inhibition of rat vas deferens smooth muscle. Of the chemical constituents of SJW tested hyperforin but not hypericin or the flavonoids quercitrin, rutin and kaempferol inhibited phenylephrine induced contractions. SJW and hyperforin also inhibited phenylephrine induced contractions in human vas deferens CONCLUSIONS: The results of our study demonstrate that SJW directly inhibits rat and human vas deferens contractility. If confirmed in vivo, these results suggest that SJW might affect sexual function in humans. These results might explain delayed ejaculation described in patients receiving SJW. PMID- 15879887 TI - Identification of pretreatment agents to enhance adenovirus infection of bladder epithelium. PMID- 15879890 TI - The urothelium in overactive bladder: passive bystander or active participant? PMID- 15879891 TI - Role of the urothelium in bladder function. PMID- 15879892 TI - Muscarinic acetylcholine receptor knockout mice: novel phenotypes and clinical implications. PMID- 15879895 TI - Re: The prostate specific antigen era in the United States is over for prostate cancer: what happened in the last 20 years? PMID- 15879896 TI - Re: Successful outpatient management of the nonpalpable intra-abdominal testis with staged Fowler-Stephens orchiopexy. PMID- 15879897 TI - Re: Immediate closure of nephrostomy tube wounds using a tissue adhesive: a novel approach following percutaneous endourological procedures. PMID- 15879899 TI - Re: Biomarkers of congenital obstructive nephropathy: past, present and future. PMID- 15879900 TI - Re: Transurethral microwave thermotherapy for benign prostate hyperplasia: separating truth from marketing hype. PMID- 15879902 TI - Re: Varicocele. Where, why and, if so, how? PMID- 15879904 TI - Swallowing dysfunction in 101 patients with nephropathic cystinosis: benefit of long-term cysteamine therapy. AB - Nephropathic cystinosis is a rare, autosomal recessive lysosomal storage disorder caused by mutations in the CTNS gene that codes for a cystine transporter in the lysosomal membrane. Affected patients store 50-100 times the normal amounts of cystine in their cells, and suffer renal tubular and glomerular disease, growth retardation, photophobia, and other systemic complications, including a myopathy and swallowing dysfunction. Using videofluoroscopy and ultrasound examinations, we assessed the swallowing function of 101 patients with nephropathic cystinosis on their most recent admission to the National Institutes of Health Clinical Center between 1987 and 2004. These patients ranged in age from 6 to 45 years; more than half had significant complaints of swallowing difficulty. On examination of barium swallow, the oral, pharyngeal, and esophageal phases of swallowing were abnormal in 24%, 51%, and 73% of patients, respectively. The frequency of dysfunction increased with age for each phase of swallowing. Both the Swallowing Severity Score (a measure of dysfunction on barium swallow) and the Oral Muscle Composite Score (a reflection of vocal strength, oral-facial movement, and tongue and lip function) increased (that is, worsened) with the number of years that a patient was not receiving treatment with cysteamine, the cystine-depleting agent of choice in cystinosis. The severity scores decreased with the number of years on cysteamine therapy. The Swallowing Severity Score varied directly with the severity of muscle disease, but was not correlated with the presence or absence of the 57-kb CTNS deletion that commonly occurs in nephropathic cystinosis patients. We conclude that swallowing dysfunction in cystinosis presents a risk of fatal aspiration, correlates with the presence of muscle atrophy, and, based on cross-sectional data, increases in frequency with age and number of years without cysteamine treatment. Cystine-depleting therapy with cysteamine should be considered the treatment of choice for both pre- and posttransplant cystinosis patients. PMID- 15879905 TI - Bacteremic pneumococcal pneumonia: a prospective study in Edmonton and neighboring municipalities. AB - Bacteremic pneumococcal pneumonia (BPP) is an important disease that should be frequently re-evaluated due to changes in demographics and recommended treatment. We conducted a prospective study from 2000 to 2002 in adults aged 17 years and over who presented to any of 6 hospitals and 1 freestanding emergency room in Edmonton, Alberta, with signs and symptoms compatible with pneumonia, a chest radiograph interpreted as pneumonia by the attending physician, and a positive blood culture for Streptococcus pneumoniae. We identified 129 patients with BPP, for an overall incidence of 9.7/100,000 person years. The rate was markedly higher among pregnant women, homeless persons, and those in prison. Sixteen percent were managed as outpatients, 61.2% as ward patients, and 22.5% required admission to the intensive care unit (ICU). Tobacco smoking was predictive of BPP, and antibiotic therapy before presentation was protective. According to pneumonia severity index, 47.3% were in low-risk classes I-III, 31.0% were in class IV, and 21.7% were in class V. Twelve (9.3%) patients died. Four died within 24 hours of arrival at hospital, and 2 had end-stage lung disease that resulted in a decision to discontinue therapy. Of the S. pneumoniae isolates, 12.5% were not susceptible to penicillin. The overall rate of BPP appears to be decreasing, although the rate is markedly increased in certain populations, which now should be targeted for vaccination. We identified 3 subsets of patients with BPP according to the site of care (ambulatory, ward, and ICU), with different outcomes. PMID- 15879906 TI - Blood culture-negative endocarditis in a reference center: etiologic diagnosis of 348 cases. AB - To identify the current etiologies of blood culture-negative infective endocarditis and to describe the epidemiologic, clinical, laboratory, and echocardiographic characteristics associated with each etiology, as well as with unexplained cases, we tested samples from 348 patients suspected of having blood culture-negative infective endocarditis in our diagnostic center, the French National Reference Center for Rickettsial Diseases, between 1983 and 2001. Serology tests for Coxiella burnettii, Bartonella species, Chlamydia species, Legionella species, and Aspergillus species; blood culture on shell vial; and, when available, analysis of valve specimens through culture, microscopic examination, and direct PCR amplification were performed. Physicians were asked to complete a questionnaire, which was computerized. Only cases of definite infective endocarditis, as defined by the modified Duke criteria, were included. A total of 348 cases were recorded-to our knowledge, the largest series reported to date. Of those, 167 cases (48%) were associated with C. burnetii, 99 (28%) with Bartonella species, and 5 (1%) with rare, fastidious bacterial agents of endocarditis (Tropheryma whipplei, Abiotrophia elegans, Mycoplasma hominis, Legionella pneumophila). Among 73 cases without etiology, 58 received antibiotic drugs before the blood cultures. Six cases were right-sided endocarditis and 4 occurred in patients who had a permanent pacemaker. Finally, no explanatory factor was found for 5 remaining cases (1%), despite all investigations.Q fever endocarditis affected males in 75% of cases, between 40 and 70 years of age. Ninety-one percent of patients had a previous valvulopathy, 32% were immunocompromised, and 70% had been exposed to animals. Our study confirms the improved clinical presentation and prognosis of the disease observed during the last decades. Such an evolution could be related to earlier diagnosis due to better physician awareness and more sensitive diagnostic techniques. As for Bartonella species, B. quintana was recorded more frequently than B. henselae (53 vs 17 cases). For 18 patients with Bartonella endocarditis, the responsible species was not identified. Species determination was achieved through culture and/or PCR in 49 cases and through Western immunoblotting in 22. Comparison of B. quintana and B. henselae endocarditis revealed distinct epidemiologic patterns. The 2 cases due to T. whipplei reflect the emerging role of this agent as a cause of infective endocarditis. Because identification of the bacterium was possible only through analysis of excised valves by histologic examination, PCR, and culture on shell vial, the prevalence of the disease might be underestimated. Among patients who received antibiotic drugs before blood cultures, 4 cases (7%) were found to be associated with Streptococcus species (2 S. bovis and 2 S. mutans) through 16S rDNA gene amplification directly from the valve, which shows the usefulness of this technique in overcoming the limitations of previous antibiotic treatment. Right-sided endocarditis occurred classically in young patients (mean age, 36 yr), intravenous drug users in 50% of cases, and suffering more often from embolic complications. Finally, 5 cases without etiology or explaining factors were all immunocompetent male patients with previous aortic valvular lesions, and 3 of the 5 presented with an aortic abscess. Further investigations should be focused on this group to identify new agents of infective endocarditis. PMID- 15879907 TI - Brucellosis in San Diego: epidemiology and species-related differences in acute clinical presentations. AB - Although aggressive public health measures have greatly reduced the number of brucellosis cases in the United States, there is a resurgence of interest in this worldwide zoonosis because of its potential as a bioweapon and its 8-fold higher incidence in California, Texas, and the other borderlands between the United States and Mexico compared with the national rate. Accordingly, we reviewed the clinical records of 28 patients diagnosed at a university hospital in San Diego, CA, between 1979 and 2002 to look for new epidemiologic trends and to test the hypothesis that there are species-specific differences in clinical presentations. In contrast to the latest California-wide study completed in 1992, Brucella abortus infections were more common (73%) than Brucella melitensis after 1992, and women were more commonly infected (77% compared with 39%) than men. Major risk factors remained Hispanic ethnicity, travel to Mexico, and ingestion of nonpasteurized dairy products. Analysis of diagnostic procedures suggested that the traditional practice of prolonged incubation of blood cultures increased their sensitivity for Brucella, even in automated radiometric systems. Direct comparison of the clinical manifestations of infections with B. abortus and B. melitensis strongly supported differences in acute presentations. B. melitensis presented more acutely as fevers of unknown origin with statistically significant higher rates of abdominal tenderness, hepatomegaly, splenomegaly, thrombocytopenia, pancytopenia, and hepatic dysfunction. These results suggest that the epidemiology of brucellosis in California may be evolving, and they show, to our knowledge for the first time in a single series, that species specific differences in presentations may account for some of the protean manifestations of brucellosis. Familiarity with manifestations of brucellosis and the optimal laboratory techniques for its diagnosis could help physicians protect the public against this reemerging, under-recognized zoonosis. PMID- 15879908 TI - Apoplexy in pituitary macroadenoma: eight patients presenting in 12 months. AB - Pituitary apoplexy is an ill-defined clinical entity. Some authors include hypoxic pituitary infarction, even in the absence of tumor after hemorrhagic delivery, whereas others apply this term strictly to hemorrhage within a pituitary adenoma. We conducted the present study to establish the prevalence, clinical characteristics, and outcome of pituitary apoplexy, defined as an endocrine crisis characterized by acute intense headache, with or without altered consciousness, rapid development of visual or motor ocular disorders, and pituitary failure, associated with a large pituitary adenoma. We describe 8 consecutive patients (1 woman and 7 men, aged 29-66 yr) presenting over 12 months with pituitary apoplexy. We reviewed patient charts for symptoms, imaging characteristics, hormonal data, management, pathologic findings, and outcome. We examined our pituitary tumors database for cases of macroadenoma without apoplexy occurring during the same period. In 5 patients, potential precipitating factors were present. In 6 patients (3 nonsecreting tumors, 1 free-alpha-subunit secreting tumor, 1 growth hormone and prolactin-secreting tumor with acromegaly, and 1 prolactinoma), no pituitary disease was suspected before the acute event, representing 19% of newly diagnosed pituitary macroadenomas during the same period of time, a higher proportion than expected from our previously published series. The 2 other patients had known pituitary macroadenomas, a nonsecreting tumor and a prolactinoma on dopamine agonist therapy. Pituitary insufficiency at diagnosis included adrenal failure in 4 patients. Transsphenoidal tumor removal was performed 3-9 days after the onset of symptoms (mean, 5.3 d) in 7 of the 8 patients. Pathologic analysis disclosed tumor hemorrhage in 4 cases, ischemic necrosis in 2, and ischemia after intrasellar hemorrhage in 1. Preoperative magnetic resonance imaging was more sensitive than computed tomography for identifying hemorrhage. The newly diagnosed prolactinoma was treated with dopamine agonist. Complete neuro-ophthalmic recovery was observed in all cases, but only 2 patients displayed normal pituitary function on follow-up. The other 6 patients required long-term hormone replacement therapy. These data show that early surgical decompression prevents persistent neuro-ophthalmic deficit, but does not prevent persistent pituitary insufficiency. Moreover, published data indicate that the efficacy of surgery for the relief of neuro-ophthalmic symptoms decreases with increasing syndrome duration. Our data confirm that apoplexy occurs most often as the inaugural manifestation of pituitary macroadenoma, and suggest a recent increase of cases of apoplexy in our area. PMID- 15879909 TI - Cross-cultural vasomotor symptom reporting: conceptual and methodological issues. PMID- 15879910 TI - Anxiety, hormonal changes, and vasomotor symptoms during the menopause transition. PMID- 15879911 TI - The forties. PMID- 15879912 TI - Recalling age at menopause. PMID- 15879913 TI - Vasomotor symptom prevalence and language of menopause in Japan. AB - OBJECTIVE: To characterize the prevalence of vasomotor symptoms affecting menopausal women in Japan, the relationship between vasomotor symptoms and menopause status as defined by menstrual patterns versus self-assigned konenki (climacteric) status, and the salience and specificity of hot flash terminology. DESIGN: Participants were healthy Japanese women, aged 45 to 55 years, living in Kyoto and Fukushima prefectures, divided into menopause groups based on menstrual patterns and self-assigned konenki groups. Women recalled 82 general health symptoms during the previous 2 weeks, and provided detailed descriptions of hot flash symptom terms. RESULTS: Hot flash prevalence ranged from 3.0% (hotto furasshu) to 17.1% (hoteri) depending on the individual term used, and was 22.1% when all hot flash terms were combined. Prevalence of vasomotor symptoms was 25.7%, but peaked at 52.6% among late perimenopausal women. Hot flash prevalence was 13.2% among premenopausal women, but zero among self-assigned pre-konenki women. The three major hot flash terms (kaa, nobose, and hoteri) vary in their physical location, characteristics, and cultural salience. CONCLUSIONS: Vasomotor symptom prevalence rates in Japan have increased over the past 20 years but are still lower than in most Western populations, and vary according to hot flash term, and between menopause and self-assigned konenki status. Konenki status appears to be a better predictor of hot flashes than menopause status. Japanese women's language, including self-assessed konenki stage and hot flash terminology, seems to be more fine-grained than Western biomedical terms of menopause status and hot flash, and may offer insight into subtle differences in experiences and underlying physiology of menopause. PMID- 15879914 TI - The role of anxiety and hormonal changes in menopausal hot flashes. AB - OBJECTIVE: To estimate the association of anxiety with menopausal hot flashes in the early transition to menopause. DESIGN: A randomly identified, population based cohort of midlife women followed up for 6 years to assess reproductive hormones and other physical, emotional, and behavioral factors. At enrollment, the women were premenopausal, aged 35 to 47 years, and had regular menstrual cycles in the normal range. Enrollment was stratified to obtain equal numbers of African American (n = 219) and white (n = 217) women. RESULTS: At the 6-year endpoint, 32% of the women were in the early transition stage and 20% reached the late menopausal transition or were postmenopausal. Reports of hot flashes increased with the transition stages, which were determined by bleeding patterns. At endpoint, hot flashes were reported by 37% of the premenopausal women, 48% of those in the early transition, 63% of women in the late transition, and 79% of the postmenopausal women. Anxiety scores were significantly associated with the occurrence of hot flashes and were also significantly associated with the severity and frequency of hot flashes (each outcome at P < 0.001). Compared with women in the normal anxiety range, women with moderate anxiety were nearly three times more likely to report hot flashes and women with high anxiety were nearly five times more likely to report hot flashes. Anxiety remained strongly associated with hot flashes after adjusting for menopause stage, depressive symptoms, smoking, body mass index, estradiol, race, age, and time. In a predictive model, anxiety levels at the previous assessment period and the change in anxiety from the previous assessment period significantly predicted hot flashes (P < 0.001). CONCLUSIONS: Anxiety is strongly associated with menopausal hot flashes after adjusting for other variables including menopause stage, smoking, and estradiol levels. Anxiety preceded hot flashes in this cohort. Additional studies are needed to examine the duration of menopausal hot flashes and to determine whether treatments that target anxiety effectively reduce menopausal hot flashes. PMID- 15879915 TI - The relationship between the endocrine characteristics and the regularity of menstrual cycles in the approach to menopause. AB - OBJECTIVE: There is currently little longitudinal data available on the serum hormonal characteristics of the menstrual cycles observed in women as they approach their final menstrual period (FMP) or menopause. We sought to determine whether the onset of irregular menses, marking the menopause transition, signifies the occurrence of anovulatory, potentially infertile cycles. DESIGN: We studied 12 subjects, initially aged 45 to 47 years, who provided daily menstrual diaries, and had blood samples collected annually, three times weekly for 4 consecutive weeks, over a period of 36 to 98 months until FMP, for measurements of serum follicle-stimulating hormone (FSH), luteinizing hormone, estradiol, and progesterone. The definition of entry into the early menopause transition was the occurrence of more than two cycles, in any consecutive sequence of 10, where cycle length was less than 23 or more than 35 days. Entry into the late transition was determined from the first observation of either 60-day or 90-day amenorrhea. Cycles were characterized endocrinologically as normal ovulatory, abnormal luteal phase, and anovulatory with evidence of ovarian follicular activity. RESULTS: The early transition had an average duration of 47 months from onset until FMP. Ten of the 12 subjects had one or more ovulatory cycles during the transition. Anovulatory cycles with ovarian activity were noted in 9 of the 12 subjects, only after entry into early and/or late transition. CONCLUSIONS: Ovulatory cycles occurred both before and after entry into the early and/or late menopause transition in subjects older than 45 years of age, whereas anovulatory cycles were observed only during the transition. The ovulatory cycles were generally associated with normal menses, whereas anovulatory cycles showed long duration and/or abnormal bleeding patterns. The occurrence of cycle irregularity is associated with an increasing frequency of anovulatory cycles, which herald the occurrence of FMP. No conclusion could be drawn regarding the appropriate definition of entry into the late transition. The definition adopted for entry into the early transition merits further validation. PMID- 15879916 TI - Reproducibility of self-reported menopause age at the 24-year follow-up of a population study of women in Goteborg, Sweden. AB - OBJECTIVE: To study the reproducibility of self-reported age at natural menopause, reported by women in their 70s, compared with menopause age reported in the initial postmenopausal period. DESIGN: A prospective study conducted in Goteborg, Sweden, based on a random sample of the total female population, started in 1968-1969, with follow ups in 1974-1975, 1980-1981, and 1992-1993. A total of 1,009 women born in 1922, 1918, or 1914 (participation rate 90.3%), representative of women of the respective ages in the general population, participated in the initial examination in 1968-1969. Of those women reporting natural menopause some time between 1968 and 1981, 565 women recalled their menopause age in 1992. RESULTS: The mean interval between the first and second reports was 18.3 years. The mean difference between first-reported and recalled menopause was 0.05 years. When menopause age was recalled in 1992-1993, 55.6% of the women with a natural menopause recalled their age at menopause correctly within 1 year, 22.6% underestimated their menopause age by more than 1 year, and 21.8% overestimated their menopause age by more than 1 year. Women undergoing early menopause (<45 years) tended to overestimate menopause age, whereas women undergoing late menopause (>55 years) tended to underestimate menopause age. These differences could not be explained by age, interval since menopause, smoking, exercise, education, or socioeconomic status. CONCLUSIONS: Menopause ages reported at an interval of almost 20 years showed a significant correlation. However, a regression tendency of the values toward the mean was observed, suggesting that strong correlation at the group level does not imply precision at the extremes. PMID- 15879917 TI - Sequential classification of endocrine stages during reproductive aging in women: the FREEDOM study. AB - OBJECTIVE: Reproductive aging involves complex endocrine changes affecting women's fertility, health, and well-being; however, understanding of the specific changes involved is limited by the lack of detailed quantitative studies. We undertook a thorough study with the aim of characterizing the different endocrine stages involved in female reproductive aging. DESIGN: FREEDOM is a cohort study designed to determine the endocrine changes during reproductive aging in women. Here, we ascertained the different endocrine patterns in a representative population and developed a staging system. In this study, 112 women aged 30 to 58 years collected daily urine samples over a 6- to 18-month period and recorded their menstrual periods. A total of 36,786 samples were analyzed for follicle stimulating hormone (FSH), luteinizing hormone, estrone 3-glucuronide, and pregnanediol 3-glucuronide. RESULTS: A classification of five sequential endocrine stages of reproductive aging was developed: stage 1, regular menstrual cycles with mean initial (day 1-5) FSH less than 5 IU/L; stage 2, regular cycles with FSH greater than 5 IU/L; stage 3, menstrual irregularity (with the appearance of "delayed-response cycles"); stage 4, acyclical ovarian activity with no evidence of ovulation and luteinization; and stage 5, ovarian quiescence and persistently raised gonadotropins. Distinct hormonal characteristics during the follicular and luteal phase were noted at each stage. CONCLUSION: This classification provides a detailed insight into the endocrinology of reproductive aging in women that could be useful for both clinical guidance and personal health care. PMID- 15879918 TI - Dietary soy protein and isoflavones have no significant effect on bone and a potentially negative effect on the uterus of sexually mature intact Sprague Dawley female rats. AB - OBJECTIVE: To evaluate the effect of dietary soy protein and isoflavones on bone and the reproductive tract in premenopausal rats. DESIGN: Three-month-old intact Sprague-Dawley female rats (N = 50) were fed diets containing casein, soy protein, or casein with isoflavone extract for 12 weeks. The amount of casein, soy protein, and extract (per kilogram diet) in each group was: (1) 200 g casein (control); (2) 100 g casein plus 100 g soy protein (low soy); (3) 200 g soy protein (high soy); 4) 200 g casein plus 17.2g extract (low extract); and (5) 200 g casein plus 34.4 g extract (high extract). Diet consumption, body weight, uterine wet weight, urinary deoxypyridinoline concentration, and bone mineral density of the femur and lumbar vertebrae were measured. Femur rigidity was evaluated by histomorphometry. The uterus and vagina were studied histologically. RESULTS: Rats in all treatment groups had lower body weights and lower deoxypyridinoline concentrations compared with controls, but none of the differences was statistically significant. There was no significant difference in femur and lumbar bone mineral density, uterine wet weights, or histomorphometry between the control and treatment groups. Histologically, uteri and vaginae were normal in all groups except that 1 of 10 rats in the high-soy group and 2 of 10 rats in the high-extract group showed extensive squamous metaplasia in the uterine gland. CONCLUSION: These results suggest that dietary isolated soy protein and isoflavones have no effect on bone and the vagina during premenopausal period, but may have an adverse effect on the uterus. PMID- 15879919 TI - Effects of soy isoflavones on endothelial function in healthy postmenopausal women. AB - OBJECTIVE: To evaluate the effects of soy isoflavone administration on endothelial function in healthy postmenopausal women. DESIGN: Sixty naturally postmenopausal women were randomly assigned to receive isoflavone or placebo tablets for 6 months. Endothelium-dependent vasodilatation was measured by brachial reactivity technique along with levels of plasma soluble intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E selectin, P-selectin and soluble thrombomodulin, von Willebrand factor, and tissue plasminogen activator. Differences between endothelium-dependent and endothelium-independent vasodilatation were assessed by evaluating brachial reactivity parameters after reactive hyperemia and after sublingual administration of nitroglycerin; furthermore, in the active group, the effect of isoflavones was also evaluated during the intra-arterial infusion of N-monomethyl L-arginine. Serum levels of lipids [high-density lipoprotein and low-density lipoprotein cholesterol, triglycerides and lipoprotein(a)] and hemostatic factors (prothrombin, fibrinogen, plasminogen activator inhibitor-1, and fibrin D-dimer) were also measured. To confirm the absorption of isoflavones, their blood concentrations were determined. RESULTS: Isoflavone treatment versus placebo was associated with a significant improvement in endothelium-dependent vasodilatation but had no impact on endothelial-independent arterial diameter and flow. Intra arterial infusion of N-monomethyl-L-arginine inhibited the significant effect of isoflavones on endothelium-mediated vasodilatation. Furthermore, isoflavone group experienced statistically significant reductions in plasma concentrations of ICAM 1, VCAM-1, and E-selectin. Levels of soluble thrombomodulin, von Willebrand factor, tissue plasminogen activator, lipids, and hemostatic factors did not change significantly throughout the study in both groups. CONCLUSIONS: Our findings suggest a positive influence of soy isoflavones on endothelial function in healthy postmenopausal women as evidenced by an improvement in endothelium dependent vasodilatation and a reduction in plasma adhesion molecule levels. PMID- 15879920 TI - A longitudinal study of the predictors of prevalence and severity of symptoms commonly associated with menopause. AB - OBJECTIVE: To delineate the role of hormone levels, menopause status, exogenous hormone use, and personal characteristics in the changing prevalence and impact of menopause symptoms. DESIGN: Annual longitudinal data were from Michigan Bone Health Study enrollees aged 24 to 44 years at baseline and followed up for a 10 year period beginning in 1992. In self-administered interviews, women reported the presence of and degree of bother (values from 1 = low to 8 = high) for symptoms related to sexuality, vasomotor, sleep/fatigue, negative mood, hair/skin, and urinary problems. Annually, collected sera samples were analyzed for estradiol, follicle-stimulating hormone, and testosterone concentrations. RESULTS: Increasing age consistently predicted the development and bother of the measured symptoms. Transition to postmenopause and higher levels of follicle stimulating hormone were significantly predictive only of the sexuality and vasomotor constructs, whereas higher estradiol levels were protective against increasing frequency and bother from sexuality and vasomotor constructs. Problems with sleeping, and secondarily, vasomotor symptoms, were the most bothersome constructs. Higher body mass index and current smoking behavior were highly related to increased bother with many symptom constructs, but especially vasomotor symptoms. Exogenous hormone use was associated with more bother from all symptom constructs. CONCLUSIONS: Sexuality and vasomotor symptom constructs seem to be more related to menopause than other constructs. The frequency of other constructs in the pre-menopause and their very strong association with increasing age suggest caution in attributing these factors directly to neuroendocrine events of the menopause transition. The strong associations between smoking and body size with symptoms, particularly vasomotor symptoms, suggest that interventions directed at these personal characteristics might be effective in dampening their impact. PMID- 15879921 TI - Prevalence of urinary incontinence in relation to self-reported depression, intake of serotonergic antidepressants, and hormone therapy in middle-aged women: a report from the Women's Health in the Lund Area study. AB - OBJECTIVE: To study a perceived relationship of urinary incontinence with self reported depression among middle-aged women, and to outline whether hormone therapy, selective serotonin reuptake inhibitors, or serotonin norepinephrine reuptake inhibitors influence the prevalence of urinary incontinence. DESIGN: A population-based cross-sectional study of 6,642 women from the Lund area, Sweden, aged 50 to 64 years, who answered questions on urinary incontinence. We obtained information on urinary incontinence, self-reported depression, and medication by questionnaire and personal interviews between December 1995 and February 2000. RESULTS: Thirty-one percent of the women reported urinary incontinence of various severities and 52% reported episodes of depression. Urinary incontinence and self reported depression were significantly associated [odds ratio (OR) 1.37; 95% CI, 1.23-1.52; P < 0.001]. Among women with self-reported depression, the use of serotonergic antidepressants was statistically associated with urinary incontinence (OR 1.78; 95% CI, 1.33-2.38; P < 0.001). Among women not reporting depression, hormone therapy was significantly associated with urinary incontinence (OR 1.45; 95% CI, 1.22-1.70; P < 0.001). After adjustment for other potential risk factors, serotonergic antidepressants and hormone therapy use were no longer associated with urinary incontinence in women with self-reported depression. However, among women not reporting depression, hormone therapy was still significantly more common in women with urinary incontinence. CONCLUSIONS: Urinary incontinence and self-reported depression seemed to be associated in middle-aged women. Use of serotonergic antidepressants or hormone therapy does not seem to be associated with a lower prevalence of urinary incontinence. PMID- 15879922 TI - Polymorphisms associated with thrombophilia and vascular homeostasis and the timing of menarche and menopause in 728 white women. AB - OBJECTIVE: Genetic factors have been proposed as modulators of the timing of natural menopause. Single nucleotide polymorphisms (SNPs) of genes associated with thrombophilia and vascular homeostasis may interfere with ovarian function and thus are candidate genes for early menopause. We attempted to assess the association of SNPs and the timing of menarche and natural and surgical menopause in an ethnically homogenous cohort of Middle European white women. DESIGN: In the present cross-sectional study, eight SNPs of six genes involved in vascular function and homeostasis were analyzed by sequencing-on-chip using solid-phase polymerase chain reaction on oligonucleotide microarrays in 728 white women: factor V (F5) Leiden G1691A, factor II (F2) prothrombin G20210A, plasminogen activator inhibitor1 (PAI-1) 4G/5G, angiotensinogen (AGT) Met235Thr, endothelial nitric oxide synthase (NOS3) T768C and NOS3 Glu298Asp, apolipoprotein E-1 (APOE 1) Cys112Arg, and APOE-2 Arg158Cys. The women's reproductive and medical histories were correlated to genotypes. RESULTS: In a univariate analysis, current smoking (P = 0.01) and the presence of at least one mutant allele of F5 Leiden (P = 0.03) and APOE-2 (P = 0.03) were significantly associated with a reduced age at natural menopause. The presence of at least one mutant allele of F5 Leiden (P = 0.02) and a body mass index above 25 kg/m (P = 0.009) were significantly associated with an increased risk for surgical menopause by premenopausal hysterectomy (odds ratio = 2.6, 95% CI, 1.2-5.6; odds ratio = 1.9, 95% CI, 1.2-3.0, respectively). Age at menarche was not affected by the carriage of any of the investigated SNPs. Applying stepwise linear regression models considering all two-way interactions, no interactions were found among different SNPs or between SNPs and environmental and lifestyle parameters. CONCLUSION: We identified various genetic and personal history parameters influencing age at natural menopause and the risk of undergoing premenopausal hysterectomy. To the best of our knowledge, we present the largest study to date determining SNPs as contributors to the genetic control of the timing of natural and surgical menopause. PMID- 15879923 TI - Bone mineral density in postmenopausal women treated with a vaginal ring delivering systemic doses of estradiol acetate. AB - OBJECTIVE: To evaluate the effect on bone mineral density of vaginal rings delivering estradiol acetate at two systemic doses versus a locally active vaginal ring in healthy postmenopausal women. DESIGN: A total of 174 postmenopausal women (younger than age 65 years) were randomly assigned to a 0.05 mg/day vaginal ring, 0.1 mg/day vaginal ring, or 0.0075 mg/day vaginal ring (active comparator), and treated for 96 weeks. Of these, 170 took a study drug; 85 taking the study drug had data at 96 weeks, and 132 women were included in the intent-to-treat analysis. Non-hysterectomized women received 1 mg of norethisterone taken on the last 12 days of each 28-day monthly cycle. The primary endpoint was change in lumbar spine bone mineral density (L2-L4); change in total hip bone mineral density was a secondary endpoint. RESULTS: At 96 weeks, mean lumbar spine bone mineral density increased 2.7% and 3.3% from baseline, respectively, in the 0.05-mg and 0.1-mg groups (P < 0.001 for both) compared with an 0.3% increase in the 0.0075-mg group (P = 0.56). Mean total hip bone mineral density increased 1.7% and 1.8% from baseline, respectively, in both the 0.05-mg and 0.1-mg groups (P < 0.001) and decreased 1.2% in the 0.0075-mg group (P = 0.001). All vaginal ring doses were well tolerated. CONCLUSIONS: Vaginal rings delivering systemic doses of estradiol increase bone mineral density of the lumbar spine and total hip in healthy postmenopausal women. Safety and acceptability were similar to existing estradiol therapies. PMID- 15879924 TI - Endothelium-dependent dilatory effects of 3alpha-OH-tibolone in gracilis muscle arterioles of the female Wistar rat. AB - OBJECTIVE: Tibolone is a synthetic steroid used for the treatment of the symptoms of menopause that, once metabolized, has estrogenic, progestogenic, and androgenic properties. We investigated the direct vasodilatory effects of the major active tibolone metabolite 3alpha-OH-tibolone and its sulfated form on female rat skeletal muscle arterioles, which play an important role in the control of blood pressure. DESIGN: In isolated, pressurized spontaneously constricted arterioles (mean passive diameter 83 +/- 3 microm), we investigated the vasodilatory effect of 3alpha-OH-tibolone and its sulfated form. To study the role of the endothelium and in particular that of nitric oxide, we repeated the experiments with 3alpha-OH-tibolone after removal of the endothelium and on vessels pretreated with the nitric oxide synthesis inhibitor, Nomega-nitro-L arginine (L-Na). Finally we compared the vasodilatory effect of 3alpha-OH tibolone with 17beta-estradiol. RESULTS: A dose-dependent dilatation to 3alpha-OH tibolone was observed starting at a concentration of 10 M. With the sulfated form of 3alpha-OH-tibolone, dilatation was only present at the highest concentration (10 M). In the denuded vessels, the vasodilatory effect was absent at concentrations from 10 to 10 M. The dilatation induced by 3alpha-OH-tibolone was not significantly reduced by L-Na. The vasodilatory effect of 3alpha-OH-tibolone did not differ from that of 17beta-estradiol. CONCLUSIONS: 3alpha-OH-tibolone has a dose-dependent vasodilatory effect on isolated skeletal muscle arterioles from the rat. The sulfated form has no vasodilatory effect in this setup. This finding suggests that during this short incubation time there was no conversion of the sulfated metabolite into its active form by the vascular endothelium. The vasodilatory effect of 3alpha-OH-tibolone is endothelium dependent at physiologic concentrations and comparable to that of 17beta-estradiol. PMID- 15879925 TI - Menopause symptoms in HIV-infected and drug-using women. AB - OBJECTIVE: To examine the association of HIV infection, drug use, and psychosocial stressors with type and frequency of menopause symptoms. DESIGN: In a cross-sectional study, HIV-infected and HIV-uninfected midlife women underwent standardized interviews on menopause status and symptoms, demographic characteristics, depressive symptoms, negative life events, and substance abuse. Body mass index (BMI), HIV serostatus, and CD4 count were measured. Associations between study variables and menopause symptoms were assessed using generalized estimating equations. RESULTS: Of 536 women not on hormone therapy, 48% were black, 42% were Hispanic, 54% were HIV positive, and 30% recently had used illicit drugs. The mean age was 45 +/- 5 years; 48% of the women were identified as premenopausal, and 37% were perimenopausal. Psychological symptoms were most prevalent (89%), followed by arthralgias (63%) and vasomotor symptoms (61%). Perimenopausal women reported significantly more menopause symptoms than premenopausal women (ORadj 1.34, 95% CI, 1.09-1.65). HIV-infected women were more likely to report menopause symptoms than uninfected women (ORadj 1.24, 95% CI, 1.02-1.51). Among HIV-infected women not on highly active antiretroviral therapy, symptoms decreased as the CD4 count declined. Increased menopause symptoms were significantly associated with depressive symptoms (ie, Center for Epidemiologic Studies Depression scale score > 23, ORadj1.82, 95% CI, 1.46-2.28), and with experiencing more than three negative life events (ORadj 2.08, 95% CI, 1.54 2.81). Increasing BMI (per kg/m) was also associated with more menopause symptoms (ORadj 1.03, 95% CI, 1.02-1.05). CONCLUSION: HIV-infected women reported more menopause symptoms than HIV-uninfected women, but symptoms were less frequent in women with more advanced HIV disease. Depressive symptoms and negative life events were also highly associated with symptoms. Further study of menopause symptoms and HIV-related factors is warranted. Mental health interventions may also have a role in ameliorating menopause symptoms. PMID- 15879938 TI - Using palliative pain care to keep a patient active. PMID- 15879939 TI - Mounting a defense against Burkholderia cepacia. PMID- 15879940 TI - New help for old bones. PMID- 15879941 TI - Inserting a nasogastric tube. PMID- 15879945 TI - What causes postmenopausal bleeding? PMID- 15879946 TI - Myths & facts...about polycythemia vera. PMID- 15879966 TI - Tailor your I.V. insertion techniques special populations. PMID- 15879969 TI - Diabetes has taken a toll on your patient's vision--how can you help? PMID- 15879970 TI - Using negative-pressure wound therapy for positive results. AB - Learn to use the system to help your patient heal. PMID- 15879971 TI - Reentering the workforce confidently. PMID- 15879972 TI - Is your patient's cancer under control? PMID- 15879973 TI - Turn back the tide of cardiogenic pulmonary edema. PMID- 15879975 TI - Grace under fire. PMID- 15879976 TI - What is necrobiosis lipoidica? PMID- 15879977 TI - Zapping VAP with evidence-based practice. PMID- 15879981 TI - Prealbumin is best for nutritional monitoring. PMID- 15879983 TI - How biometrics keep sensitive information secure. PMID- 15879984 TI - Venomous snakebite. PMID- 15879985 TI - Advanced directives: the conversation we all must have. PMID- 15879989 TI - Beyond the midpoint with continuous performance review. AB - JCAHO's new continuous periodic performance review allows organizations to assess performance throughout the year. PMID- 15879990 TI - What to know about duty to report. AB - Recognize what conduct needs to be reported to meet ethical and legal duties and to avoid ramifications. PMID- 15879991 TI - Excise the dysfunction from your executive team. AB - Healthcare organizations need to invest time in determining how to build their collection of stars into a cohesive team. PMID- 15879992 TI - Five concepts for creating change. AB - These guidelines can help new nurse managers adjust to their roles and create climates of growth and change. PMID- 15879993 TI - Balance is more than just a buzzword. AB - Before nurse leaders can create a work/life balance, they must first define their ideal scenario. PMID- 15879994 TI - Thromboelastography: a means to transfusion reduction. AB - Consider using thromboelastography to improve care outcomes for cardiac surgery patients. PMID- 15879996 TI - More than a title. AB - In an American Board of Nursing Specialties survey, nurse managers share their perceptions of the value of specialty nursing certification. PMID- 15879997 TI - Fight or flight? Disruptive behavior in medical/surgical services. AB - The implementation of a clinical nurse specialist helped staff effectively manage disruptive patient behaviors and improve morale within an acute medical/surgical service. PMID- 15879998 TI - Get a hold on costs and safety with securement devices. AB - Evidence shows that commercially available catheter securement devices both reduce accidental needlesticks to healthcare workers and prevent catheter-related bloodstream infections by limiting catheter movement. PMID- 15880000 TI - Understanding pulmonary arterial hypertension. AB - New therapies improve the management of pulmonary arterial hypertension. PMID- 15880002 TI - Proactive teams yield exceptional care. AB - Learn how a manager and her team used nine management principles to create a proactive nursing team while designing and opening a new inpatient unit. PMID- 15880004 TI - Tips for top challenges. PMID- 15880007 TI - The context in which to better understand quality care. PMID- 15880008 TI - Variability in end-of-life care--how much is too much? PMID- 15880009 TI - Evaluating corticosteroids in the pediatric intensive care unit: a tale of apples and oranges. PMID- 15880010 TI - Food for thought. PMID- 15880011 TI - Unknown anions and gaps in medical knowledge. PMID- 15880012 TI - Stress of admission to pediatric intensive care unit on children. PMID- 15880013 TI - Heat shock protein: tracing the enemy in septic shock. PMID- 15880014 TI - New technologies in the intensive care unit: a cautionary tale. PMID- 15880015 TI - Recombinant activated factor VII: panacea or passing fad? PMID- 15880017 TI - Reduction in calcineurin inhibitor exposure and maintenance of effective immunosuppression: clinical experience with everolimus (Certican). PMID- 15880018 TI - Guest editor's introduction. PMID- 15880019 TI - Introducing everolimus (Certican) in organ transplantation: an overview of preclinical and early clinical developments. AB - Everolimus (Certican) is a novel proliferation signal inhibitor with potent immunosuppressant effects that has been shown to prevent and reverse acute rejection in preclinical models of kidney, heart, and lung transplantation. These effects have been confirmed by clinical studies in which everolimus demonstrated comparable efficacy to azathioprine and mycophenolate mofetil in heart and renal transplantation, respectively, when combined with full-dose cyclosporine (CsA; Neoral). Experimental studies have also shown that everolimus inhibits smooth muscle cell proliferation and prevents neointimal thickening and transplant arteriosclerosis. Importantly, everolimus was shown to significantly reduce both the incidence and severity of cardiac allograft vasculopathy in recipients of heart transplants, which if confirmed in long-term studies, could provide a major advantage in these patients. Use of everolimus was also associated with lower rates of cytomegalovirus infection in both indications. Studies in which everolimus was combined with reduced-exposure CsA have demonstrated significantly improved renal function when compared with full-dose regimens, with no loss of efficacy. The results of preclinical and clinical studies with everolimus thus indicate that this novel proliferation signal inhibitor will be an important addition to the armamentarium by enhancing graft survival and minimizing toxicity. PMID- 15880020 TI - Concentration-controlled everolimus (Certican): combination with reduced dose calcineurin inhibitors. AB - The calcineurin inhibitors (CNIs) have been the cornerstone of immunosuppressive therapy in recent years, though their association with dose-related nephrotoxicity is a major factor in the development of chronic allograft nephropathy in kidney transplant recipients. The novel proliferation signal inhibitor everolimus (Certican) has a mode of action distinct and complementary to the CNIs, which is thought to underlie the demonstrated synergy between everolimus and cyclosporine (CsA) in preclinical models. This finding suggests that everolimus may allow for CNI dose reduction, enabling nephrotoxicity to be minimized without compromising efficacy. Two prospective, multicenter, randomized studies have investigated the efficacy and safety of everolimus (1.5 or 3.0 mg/day) combined with reduced-exposure CsA (Neoral) in de novo renal transplant patients. In these studies, the everolimus trough level was maintained at > or =3 ng/ml with therapeutic drug monitoring and the CsA dose optimized with C(2) monitoring. The incidence of efficacy failure with this regimen was shown to be comparable with that provided by regimens employing full-dose CNIs, but with good renal function. Furthermore, everolimus (trough levels 3-8 ng/ml) plus reduced exposure CsA was associated with a marked reduction in acute rejection compared with regimens using fixed-dose everolimus or mycophenolate mofetil plus full-dose CsA. At 12 months follow-up, CsA trough level was 57% lower in these trials than in previous trials in which patients received full-dose CsA. This suggests that concentration-controlled everolimus can be successfully combined with optimized administration of CsA and that this regimen may maximize immunosuppression, whilst preserving renal function. PMID- 15880021 TI - Clinical experience with everolimus (Certican): optimizing dose and tolerability. AB - BACKGROUND: Everolimus (Certican), a novel proliferation signal inhibitor, allows calcineurin inhibitor dose reduction in transplant patients, minimizing risk of nephrotoxicity without loss of immunosuppressive efficacy. As a result of its mode of action, this class of agent (i.e., everolimus and sirolimus) is associated with certain adverse events (e.g., lymphocele, arthralgia, edema and hyperlipidemia). METHODS: Three case studies from within an everolimus Phase III trial (A2306) are presented with the aim of illustrating how treatment-related adverse events can be managed. RESULTS: The combination of everolimus with reduced-exposure cyclosporine was efficacious in these patients. One experienced mild acute rejection that was managed with steroid boluses. All have acceptable graft function during follow-up to date. Moderate lymphocele either resolved spontaneously or was easily managed by povidine-iodine instillations. One patient developed serious early lymphocele that, unusually, required surgical intervention. Everolimus dose reduction or withdrawal was not necessary to effectively manage lymphocele of any severity. A case of bilateral multiple arthralgia was effectively managed with everolimus dose reduction, but still maintained everolimus trough blood levels at approximately 3 ng/ml. Eyelid and ankle edemas were also easily managed with low dose furosemide, preferably coupled with cyclosporine and everolimus dose reduction. Hyperlipidemia responded to statin therapy. CONCLUSIONS: Everolimus-related adverse events can be effectively managed either with other treatments or, where necessary, by everolimus dose reduction, without loss of efficacy. Discontinuing everolimus therapy is generally not necessary. PMID- 15880022 TI - Clinical experience with everolimus (Certican) in elderly recipients: the "old for-old" concept. AB - BACKGROUND: With an increasing number of elderly patients now waiting for a kidney transplant, and the percentage of kidney donors over 55 years also rising, harvesting older kidneys specifically for use in older recipients is a way of extending the donor pool for renal transplantation. METHODS: Two case studies from an everolimus (Certican) Phase III trial (A2306) are presented. They illustrate clinical experience of achieving stable graft function in the "old-for old" kidney transplant program at the Ramon y Cajal Hospital in Madrid, Spain. RESULTS: The first case study demonstrates that stable graft function can be achieved in an "old-for-old" kidney transplant patient, who receives everolimus in combination with reduced-exposure cyclosporine (CsA). To optimize graft function in the long-term, CsA C(2) blood levels should be <400 ng/ml. The second case study highlights how "old-for-old" renal transplant recipients can be at risk of calcineurin-inhibitor (CNI)-induced nephrotoxicity. Here, graft function did not improve following CsA dose reduction; thus, CsA was withdrawn 1 year posttransplant. Stable graft function that is acceptable for an "old-for-old" kidney transplant was then achieved with everolimus trough blood levels of 10-15 ng/ml and low-dose prednisone. CONCLUSIONS: In these cases, everolimus was used safely and effectively in the "old-for-old" kidney transplant setting. Its combination with reduced-exposure CsA, or its facilitation of CsA withdrawal, minimized the risk of nephrotoxicity in the older renal graft. These findings need to be confirmed in larger studies specific to this population. PMID- 15880023 TI - Clinical experience with everolimus (Certican) in young renal transplant recipients. AB - BACKGROUND: Young transplant recipients benefit most from graft longevity, but are at high risk of rejection. Thus, in young recipients, the challenge is provision of effective immunosuppression resulting in good renal function while minimizing the toxicities of immunosuppressant therapy. METHODS: The following two case studies describe the clinical experience of achieving stable graft function in two young female transplant recipients of older living donor kidneys. As part of an ongoing clinical trial (A2307), immunosuppressant therapy consisted of everolimus (Certican) in combination with reduced-exposure cyclosporine (CsA) and prednisone. RESULTS: The two case studies demonstrate that rejection can be prevented and good, stable renal graft function achieved with everolimus 0.75-1.5 mg bid (long-term everolimus blood trough levels: 2.3-6.5 ng/ml in Case 1 and 3.8 9.3 ng/ml in Case 2) plus reduced-exposure CsA. CsA was reduced from an initial dose of 100 mg bid (C2 target levels of 500-700 ng/ml) to 75 mg and 35 mg bid in the two cases, respectively, in order to reach maintenance C2 target levels of 230-450 ng/ml. Maintenance prednisone dose was low in both cases (7-7.5 mg/day). Both patients tolerated the immunosuppressant regime well, and at 31-33 months follow-up they remained free of acute rejection episodes and toxicities associated with CsA. CONCLUSION: Everolimus plus reduced-dose CsA can provide safe, adequate immunosuppression resulting in prevention of acute rejection in young recipients. Reducing CsA exposure minimizes risk of nephrotoxicity and other adverse events that affect compliance in these patients for whom long-term graft survival is vital. PMID- 15880024 TI - Clinical experience with everolimus (Certican): a summary. PMID- 15880026 TI - The Global Alliance for Transplantation. PMID- 15880029 TI - Presidential address to The Transplantation Society, 2004: accomplishments, ethics, and scientific perspectives. PMID- 15880031 TI - Back to the future. PMID- 15880039 TI - Pretransplant donor peripheral blood mononuclear cells infusion induces transplantation tolerance by generating regulatory T cells. AB - BACKGROUND: It was suggested that maintenance of tolerance to organ transplantation may depend on the formation of T regulatory cells. METHODS: Lewis (LW) rats were made tolerant to a Brown Norway kidney by pretransplant donor peripheral blood mononuclear cells (PBMC) infusion. At greater than 90 days after transplantation, lymph node cells (LN) and graft-infiltrating leukocytes (GIL) alloreactivity was tested in mixed lymphocyte reaction (MLR), coculture, and transwell experiments. GIL phenotype was analyzed by FACS. mRNA expression of cytokines and other markers was analyzed on CD4+ T cells from LN. The tolerogenic potential of tolerant cells in vivo was evaluated by adoptive transfer. RESULTS: Tolerant LN cells showed a reduced proliferation against donor stimulators but a normal anti-third-party alloreactivity. In coculture, these cells inhibited antidonor but not antithird-party reactivity of naive LN cells. Interleukin (IL) 10 and FasL mRNA expression was up-regulated in tolerant CD4+ T cells, but an anti-IL-10 monoclonal antibody (mAb) only partially reversed their inhibitory effect. Immunoregulatory activity was concentrated in the CD4+ CD25+ T-cell subset. In a transwell system, tolerant T cells inhibited a naive MLR to a lesser extent than in a standard coculture. Regulatory cells transferred tolerance after infusion into naive LW recipients. CD4+ T cells isolated from tolerized grafts were hyporesponsive to donor stimulators and suppressed a naive MLR against donor antigens. CONCLUSIONS: Donor-specific regulatory T cells play a role in tolerance induction by donor PBMC infusion. Regulatory activity is concentrated in the CD4+ CD25+ subset and requires cell-to-cell contact. Regulatory CD4+ T cells accumulate in tolerized kidney grafts where they could exert a protective function against host immune response. PMID- 15880040 TI - Induction of stable mixed chimerism by embryonic stem cells requires functional Fas/FasL engagement. AB - BACKGROUND: Recent data show the efficacy of embryonic stem cells (ESC) to engraft in allogeneic recipients without host pretreatment. This property is due to their low expression of major histocompatibility complex (MHC) class I antigens and lack of MHC class II expression. Here, we tested the hypothesis that the constitutive FasL expression by ESC is a requirement for their stable engraftment in allogeneic recipients. METHODS: MRL and MRL-lpr/lpr mice (H-2k) were infused allogeneic 129SvJ RW-4 (H-2b) ESC without host preconditioning. The development of mixed chimerism was monitored over 100 days by flow cytometry. RESULTS: Mixed chimerism was detectable by day 7. The amount of donor cells detected varied between 3-5.5% and were lymphoid, but nonmyeloid. Only 50% of lpr/lpr mice engrafted and lost donor cells by day 28 post-ESC infusion. In contrast, >80% wild type mice engrafted and maintained mixed chimerism up to day 100. CONCLUSIONS: These data suggest a critical role for Fas-FasL engagement in ESC engraftment. We conclude that ESC may induce clonal deletion of alloreactive T cells by Fas-induced apoptosis in recipient T cells, protecting them from rejection. The data provide a rationale for improved protocols for the achievement of robust ESC-induced mixed chimerism. PMID- 15880041 TI - Role of the ICOS-B7h costimulatory pathway in the pathophysiology of chronic allograft rejection. AB - BACKGROUND: Inducible costimulator (ICOS) is the third member of the CD28 superfamily and has a unique role in T cell activation and function. Recent studies indicated that the ICOS-B7h pathway plays an important role in alloimmune responses. We further investigated the role of the ICOS pathway in the pathologic process of chronic rejection in vivo. METHODS: An established major histocompatibility complex class II disparate cardiac transplantation model was used. We treated mice with a blocking anti-B7h monoclonal antibody (mAb) either in the initiation phase (early blockade) or in the progression phase (delayed blockade) of disease. In addition, some mice received mAb in the entire period (whole blockade). At 6 weeks after transplantation, cardiac grafts were evaluated by histopathologic analysis in terms of vasculopathy, fibrosis, and cellular infiltration. The intragraft expressions of cytokines and chemokines were also examined by quantitative real-time polymerase chain reaction analysis. RESULTS: Early blockade of the ICOS-B7h pathway did not show any protective effect on chronic allograft rejection compared with untreated controls. In contrast, delayed blockade significantly inhibited the development of vasculopathy, fibrosis, and cellular infiltration (P=0.043, P=0.004, and P=0.03 vs. untreated control, respectively). Interestingly, whole blockade did not prevent the chronic rejection process. Furthermore, the inhibitory effect of delayed ICOS blockade on chronic rejection was associated with down-regulation of local intragraft expression of several cytokines and chemokines. CONCLUSIONS: These data suggest that the ICOS-B7h pathway is critical in the activation of effector/memory T cells that are necessary for the progression of chronic rejection and provide the rationale to develop novel and specific therapies to prevent this process. PMID- 15880042 TI - Prevention and cure of autoimmune diabetes in nonobese diabetic mice by continuous administration of FTY720. AB - BACKGROUND: Treatment of nonobese diabetic (NOD) mice with FTY720 before the development of insulitis prevents the onset of diabetes. In this study, the authors investigated whether FTY720 treatment of NOD mice with established insulitis prevents the development of diabetes. METHODS: FTY720 (1 mg/kg) was administered continuously to euglycemic NOD mice starting at 14 or 23 weeks of age. A group of untreated, age-matched NOD mice served as controls. Mice with more than 300 mg/dL blood glucose on three consecutive measurements were considered diabetic. RESULTS: Diabetes developed in control mice starting at 13 weeks of age and reached 78% by 33 weeks of age. Mice at 14 and 23 weeks of age exhibited extensive insulitis that progressed with age. Continuous oral administration of FTY720 starting at either age completely prevented the development of diabetes. However, its withdrawal at 37 weeks of age led to abrupt diabetes onset. Pancreases of FTY720-treated diabetes-free mice showed peripheral insulitis, with strong insulin staining. The protection from diabetes was also achieved by intraperitoneal injection of FTY720 or sirolimus (1.5 mg/kg). Unlike FTY720, withdrawal of sirolimus did not induce diabetes. Continuous oral FTY720 (3 mg/kg) treatment in overtly diabetic NOD mice led to complete reversal of diabetes in 6 of 11 mice. The standard adoptive transfer study in NOD-severe combined immunodeficient mice showed that peripheral lymphoid organs of FTY720 treated mice contained diabetogenic cells but not dominant immunoregulatory cells. CONCLUSIONS: FTY720, which does not cause generalized immunosuppression, may be a safe and benign therapeutic agent for chronic use to prevent or cure type 1 diabetes. PMID- 15880043 TI - Dendritic cells genetically engineered with adenoviral vector encoding dnIKK2 induce the formation of potent CD4+ T-regulatory cells. AB - BACKGROUND: Immature dendritic cells (DC), characterized by low expression of both major histocompatibility complex class II antigens and co-stimulatory molecules, can be instrumental in the induction of peripheral tolerance. Because nuclear factor (NF)-kappa B is central to DC maturation, the authors engineered DC with an adenoviral vector (Adv) encoding for a kinase-defective dominant negative form of IKK2 (dnIKK2) to block NF-kappa B activation and inhibit DC maturation. METHODS: DC were obtained by culturing bone marrow from Brown Norway (BN) rats with granulocyte-macrophage colony-stimulating factor and interleukin-4 for 11 days. To block NF-kappa B activation, at day 9, cells were transfected with AdV-dnIKK2. At day 11, cells were used as stimulators in primary mixed leukocyte reaction (MLR) with naive Lewis rat lymphocytes as responders. CD4+ T cells were purified from primary MLR and tested in secondary MLR with allogeneic mature DC and in co-culture MLR with naive lymphocytes. The tolerogenic potential of dnIKK2-DC was evaluated in vivo in a model of rat kidney allotransplantation. RESULTS: DnIKK2-DC were immature and lacked any allostimulatory activity. T cells preexposed to allogeneic dnIKK2-DC were hyporesponsive to a secondary stimulation with mature DC and acquired potent regulatory properties, inhibiting naive T-cell proliferation toward allogeneic stimuli. Pretransplant infusion of allogeneic donor dnIKK2-DC prolonged the survival of a kidney allograft from the same allogeneic donor, without the need for immunosuppressive therapy. CONCLUSIONS: Allogeneic DC, rendered immature by dnIKK2 transfection, induce in vitro differentiation of naive T cells into CD4+ T-regulatory cells, effective at low ratios with target cells, rendering them applicable for cellular therapy of immune-mediated abnormalities and for preventing transplant rejection. PMID- 15880044 TI - The granzyme B and interferon-gamma enzyme-linked immunospot assay as alternatives for cytotoxic T-lymphocyte precursor frequency after renal transplantation. AB - BACKGROUND: The interferon (IFN)-gamma enzyme-linked immunospot (ELISPOT) assay has gained increased popularity as a surrogate marker of cytotoxic T-lymphocyte (CTL) activity. However, the functional activity of CTL might be a more relevant surrogate marker of CTL. Therefore, the authors wondered whether the granzyme B (GrB) ELISPOT assay is a better marker for determining the number of CTL than the IFN-gamma ELISPOT assay. METHOD.: Peripheral blood mononuclear cells (PBMC) from 19 kidney transplant patients were stimulated with donor cells or third-party cells. The authors determined the CTL precursor frequency (CTLpf) and simultaneously measured the number of IFN-gamma- and GrB-producing cells (pc) by ELISPOT assay. RESULTS: In all three different assays, the reactivity to donor cells was significant lower than the reactivity to third-party cells: CTLpf, median: 9 versus 60/10(6) PBMC (P=0.0002); number of IFN-gamma pc: 10 versus 90/10(6) PBMC (P=0.0001); number of GrB pc: 60 versus 205/10(6) PBMC (P=0.05). When the authors compared the CTLpf after third-party stimulation with the corresponding ELISPOT results, they found a positive correlation between the CTLpf and the number of IFN-gamma pc (r(s)=0.47, P=0.05). No correlation was found between the CTLpf and the number of GrB pc (r(s)=0.23, P=0.36). However, when they compared the donor-specific CTLpf with the corresponding ELISPOT results, no correlation with the ELISPOT for IFN-gamma (r(s)=0.10, P=0.69) or GrB (r(s)=-0.24, P=0.34) was found. CONCLUSIONS: The authors feel that the CTLpf, as a measure of the actual endpoint of cytolytic activity and independent of the pathway of killing, remains the "gold standard" for determining donor-specific cytolytic activity after clinical organ transplantation. PMID- 15880045 TI - Endothelial cells derived from pigs lacking Gal alpha(1,3)Gal: no reduction of human leukocyte adhesion and natural killer cell cytotoxicity. AB - BACKGROUND: The expression of galactose-alpha(1,3)galactose (Gal) on porcine cells represents a major barrier to xenotransplantation. The generation of Gal-/- pigs to overcome this barrier redirected the focus of research to other rejection mechanisms, including cellular immunity. The present in vitro study investigated (1) the adhesive interactions between human leukocyte subsets and primary endothelial cells derived from inbred Gal-/- and Gal+/+ pigs, and (2) the susceptibility of such Gal-/- porcine endothelial cells to human natural killer (NK) cell cytotoxicity. METHODS: Primary porcine aortic endothelial cells (PAEC) were isolated from Gal-/- (PAEC-Gal-/-) and Gal (PAEC-Gal+/+) pigs. Human peripheral blood mononuclear cells (PBMC), polymorphonuclear neutrophils (PMN), and NK cells were isolated from healthy volunteers and tested in functional adhesion and cytotoxicity assays. RESULTS: Adhesion of human PBMC, PMN, or purified NK cells on PAEC-Gal-/- cells was not different from that on PAEC-Gal+/+ cells. Comparing the different leukocyte subsets of PBMC, a preferential adhesion of NK and B cells on both PAEC-Gal-/- and PAEC-Gal+/+ was detected. Tumor necrosis factor-alpha stimulation of PAEC-Gal-/- and PAEC-Gal+/+ induced an increase of CD62E and CD106 expression and increased cellular adhesion, in particular, of PMN. The lack of Gal-/- expression on PAEC-Gal cells did not prevent xenogeneic human NK-cell cytotoxicity mediated by freshly isolated or interleukin-2-activated NK cells. CONCLUSIONS: Neither human leukocyte adhesion nor xenogeneic NK-cell cytotoxicity against PAEC are impaired by the lack of Gal, indicating that Gal is not a dominant target of cellular rejection. PMID- 15880046 TI - Inhibitory and stimulatory effects of cyclosporine A on the development of regulatory T cells in vivo. AB - BACKGROUND: Regulatory T cells (Tregs) are increasingly recognized as playing a major role in nondeletional tolerance. To avoid rejection before tolerance is established, clinical trials of tolerance induction include immunosuppressive drugs early posttransplant. It is therefore essential that immunosuppressive protocols do not block Tregs generation. Tregs function has been shown to depend upon interleukin-2 signaling, but there are limited data available on how calcineurin inhibitors influence Tregs development and function in vivo. METHODS: To study this, we used a previously established rat cardiac allograft model where donor-specific Tregs and tolerance are induced by pretransplant donor-specific blood transfusion (DSBT). RESULTS: In this model, we found that adjunction of 50 mg/kg cyclosporine (CsA) (not a lower dose, 10 mg/kg) at the time of DSBT (not at the time of transplantation) abrogates Tregs development and causes rejection. Interestingly, 10 mg/kg CsA given posttransplant (day 0-11) in the absence of pretransplant DSBT induced the development of Tregs and provoked a state of tolerance indistinguishable from the one induced by DSBT. Finally, DSBT given the day of transplantation did not promote tolerance, unless recipients also received a delayed short course (day 5-9) of 10 mg/kg CsA. CONCLUSIONS: Adjunction of high dose CsA to pretransplant DSBT abrogates Tregs generation. On the contrary, a lower dose (10 mg/kg) of CsA promotes Tregs development either in synergy with perioperative DSBT (providing that a drug-free interval is respected) or by its own effect. These data provide new guidelines for a more tolerogenic use of calcineurin inhibitors in the clinic, particularly when immunomodulatory strategies aimed at inducing Tregs are applied. PMID- 15880047 TI - The CD154-CD40 T-cell co-stimulation pathway in liver ischemia and reperfusion inflammatory responses. AB - BACKGROUND: Ischemia-reperfusion (I/R) injury is a prime antigen-independent inflammatory factor in the dysfunction of liver transplants. The precise contribution of T cells in the mechanism of I/R injury remains to be elucidated. As the CD154-CD40 co-stimulation pathway provides essential second signal in the initiation and maintenance of T-cell-dependent immune responses, this study was designed to assess the role of CD154 signaling in the pathophysiology of liver I/R injury. METHODS: A mouse model of partial 90-min warm hepatic ischemia followed by 6 hr of reperfusion was used. Three animal groups were studied: (1) wild-type (WT) mice treated with Ad-(-gal versus Ad-CD40 immunoglobulin; (2) untreated WT versus CD154 (MR1) monoclonal antibody-treated WT mice; and (3) untreated WT versus CD154 knockout mice. RESULTS: The disruption of CD154 signaling in all three animal groups ameliorated otherwise fulminant liver injury, as evidenced by depressed serum glutamic oxaloacetic transaminase levels, compared with controls. These beneficial effects were accompanied by depressed hepatic T-cell sequestration, local decrease of vascular endothelial growth factor expression, inhibition of tumor necrosis factor-(and T-helper type 1 cytokine production, and induction of antiapoptotic (Bcl-2/Bcl-xl) but depression of proapoptotic (caspase-3) proteins. CONCLUSIONS: By using in parallel a gene therapy approach, pharmacologic blockade, and genetically targeted mice, these findings document the benefits of disrupting CD154 to selectively modulate inflammatory responses in liver I/R injury. This study reinforces the key role of CD154-CD40 T-cell co-stimulation in the pathophysiology of liver I/R injury. PMID- 15880048 TI - In utero bone marrow transplantation induces kidney allograft tolerance across a full major histocompatibility complex barrier in Swine. AB - BACKGROUND: In utero hematopoietic stem-cell transplantation has been shown to induce donor-specific tolerance in small-animal models. However, tolerance has been difficult to achieve in large-animal studies. METHODS: Outbred swine underwent in utero transplantation of fully major histocompatibility complex (MHC)-mismatched CD3-depleted bone marrow mixed with fresh bone marrow to achieve a final CD3 content of 1.5%. Transplantation was performed at 50 to 55 days' gestation and two animals survived long term and demonstrated multilineage peripheral blood hematopoietic chimerism. These two long-term survivors were analyzed for in vitro evidence of donor-specific tolerance by mixed leukocyte reaction (MLR), cell-mediated lysis (CML), and antibody testing and in vivo by kidney transplantation. RESULTS: Both animals demonstrated in vitro donor specific unresponsiveness by MLR and CML and did not demonstrate anti-donor antibody production. Donor matched kidney transplants were performed without immunosuppression and functioned for more than 100 days, with no evidence for rejection. CONCLUSIONS: The authors demonstrate conclusively that in utero transplantation of fully MHC-mismatched bone marrow in swine can lead to engraftment and stable multilineage hematopoietic chimerism and tolerance to postnatal donor MHC-matched kidney transplantation without the need for immunosuppression. PMID- 15880049 TI - Systemic transforming growth factor-beta1 gene therapy induces Foxp3+ regulatory cells, restores self-tolerance, and facilitates regeneration of beta cell function in overtly diabetic nonobese diabetic mice. AB - BACKGROUND: Type 1 diabetes results from auto-aggressive T-cell-mediated destruction of beta cells of the pancreas. Recent data suggest that restoration of self-tolerance may facilitate islet-cell regeneration/recovery. In view of the immunoregulatory activity of transforming growth factor (TGF)-beta1, we investigated whether systemic TGF-beta1 gene therapy blocks islet destructive autoimmunity and facilitates regeneration of beta-cell function in overtly diabetic nonobese diabetic (NOD) mice. METHODS: We used site-directed mutagenesis to create cysteine to serine mutation at sites 224 and 226 and constructed a replication deficient adenovirus (Ad) vector encoding active form of human TGF beta1 (Ad-hTGF-beta1). Overtly diabetic NOD mice received intravenous injection of Ad-hTGF-beta1. Seven to 14 days after the injection, the mice received transplants with 500 syngeneic islets under the kidney capsule. Islet-graft survival and regeneration of endogenous beta-cell function were examined. RESULTS: Syngeneic islet grafts failed by day 17 in all untreated mice, whereas Ad-hTGF-beta1 therapy prolonged survival of islet grafts. Islet grafts from treated mice showed well-preserved islets with a peri-islet infiltrate primarily of CD4+ T cells and expression of CD25 and Foxp3. Systemic TGF-beta1 gene therapy was associated with islet regeneration in the native pancreas. Native pancreas of treated mice revealed islets staining strongly for insulin. Similar to what was found in the syngeneic islet graft, there were well-demarcated peri-islet infiltrates that were positive for CD4, TGF-beta1, and Foxp3. CONCLUSIONS: Our data demonstrate that systemic TGF-beta1 gene therapy blocks islet destructive autoimmunity, facilitates islet regeneration, and cures diabetes in diabetic NOD mice. PMID- 15880050 TI - Human monocyte-derived neohepatocytes: a promising alternative to primary human hepatocytes for autologous cell therapy. AB - BACKGROUND: There is growing interest in new therapeutic options for the treatment of end-stage liver diseases. In addition to mechanical devices supporting liver function, such as bioreactors, the transplantation of hepatocyte like cells derived from (adult) stem cells offer great perspectives. We have generated hepatocyte-like (NeoHep) cells from terminally differentiated peripheral blood monocytes and, in this study, have evaluated these cells as a possible tool for autologous cell therapy. METHODS: Peripheral blood monocytes were cultured under conditions that promote hepatocyte-like differentiation and were characterized for hepatocyte marker expression by reverse-transcriptase polymerase chain reaction, immunohistochemistry, and immunoblotting and for specific secretory and metabolic functions with the appropriate biochemical assays. RESULTS: NeoHep cells resembled primary human hepatocytes with respect to morphology, expression of hepatocyte markers (albumin, cytochrome P450 isoenzymes, asialoglycoprotein receptor, coagulation factor VII), various secretory and metabolic functions (albumin secretion, urea production, lactate formation, and lactate dehydrogenase and aspartate transaminase release), and drug detoxification activities (phase I metabolization of ethoxycoumarin into 7OH coumarin after stimulation with 3-methylcholanthren, induction of CYP3A4 activity, and phase II metabolization through UDP-glucuronidation of 4-methyl umbelliferone). CONCLUSIONS: These data convincingly show that NeoHep cells display a phenotype and specific in vitro metabolic functions that are quantitatively and qualitatively comparable in part with those of primary human hepatocytes. These cells could thus be clinically applied in an autologous setting for the treatment of end-stage liver diseases or for improving liver function in patients who have undergone critical liver-mass resection. PMID- 15880051 TI - Differences in suppressor of cytokine signaling-1 (SOCS-1) expressing islet allograft destruction in normal BALB/c and spontaneously-diabetic NOD recipient mice. AB - BACKGROUND: The ability to block interferon signaling represents an important strategy in designing therapies to prevent beta-cell destruction during islet allograft rejection. METHODS: The SOCS proteins regulate cytokine signaling by blocking activation of JAK/STAT proteins. Using islets isolated from SOCS-1 transgenic mice (SOCS-1-Tg; these mice express SOCS-1 under the control of the human insulin promoter and are on the C57BL6/J background), we investigated whether SOCS proteins can prevent the destruction pancreatic islet cells transplanted beneath the kidney capsule of major histocompatibility complex mismatched normal BALB/c and spontaneously-diabetic NOD mouse recipients. RESULTS: Immunohistochemical staining for insulin confirmed the presence of donor SOCS-1-Tg islets in islet allografts harvested at 22 days posttransplant, whereas grafts of control non-Tg islets were destroyed by 14 days. In contrast, SOCS-1-Tg allogeneic islets were not protected from beta-cell destruction in clinically diabetic NOD mice. The islet allografts functioned for 1 week posttransplant; however, hyperglycemia returned after 2 weeks and the grafts were destroyed. Rejection of SOCS-1-Tg and non-Tg islets in autoimmune diabetic NOD mice was associated with an infiltrate of both CD4+ and CD8+ T cells and a T2-type cytokine response (IL-4) rather than the conventional T1-type cytokine response observed during islet allograft rejection. Self-antigen upregulation in response to IFN-gamma stimulation did not appear to be a factor in rejection of the islet allografts. CONCLUSIONS: These results demonstrate that expression of SOCS-1 in islets delays islet allograft rejection but cannot circumvent destruction of the islets by the recurrence of the tissue-specific autoimmune process of spontaneous diabetes. PMID- 15880052 TI - Dual role of vascular endothelial growth factor in hepatic ischemia-reperfusion injury. AB - BACKGROUND: Vascular endothelial growth factor (VEGF), a major angiogenic factor, mediates a variety of disease conditions through promotion of angiogenesis. It also plays a critical role as a potent proinflammatory cytokine in a variety of physiologic and pathologic immune responses. In the present study, we evaluated the expression of VEGF in hepatic warm ischemia-reperfusion (I/R) injury and examined the effect of recombinant human (rh)VEGF administration in an established murine model. METHOD: The expression of VEGF in the liver was assessed by quantitative real-time polymerase chain reaction and immunohistochemistry during I/R injury using 70% partial hepatic ischemia model. The effect of rhVEGF administration on I/R injury was evaluated by measuring liver function and histology. In addition, local inducible nitric oxide synthase (iNOS) and endothelial NO synthase expressions were examined to address the underlying mechanisms. RESULTS: The local expression of VEGF was significantly up regulated at 2 hours after reperfusion after 60 minutes of ischemia compared with that in the naive liver. VEGF was expressed predominantly in CD11b+ cells infiltrating into the ischemic liver. The administration of rhVEGF had a significant protective effect on ischemic injury in the liver. This effect was associated with the up-regulation of iNOS expression in the rhVEGF-treated liver. CONCLUSION: We demonstrate a dual role of VEGF in hepatic warm I/R injury. Although endogenous VEGF is expressed and functional to initiate hepatic I/R injury, exogenous rhVEGF has a beneficial effect on the ischemic liver. These data may provide new insights into the role of VEGF as well as pathophysiology of hepatic I/R injury. PMID- 15880053 TI - Posttransplant interleukin-4 treatment converts rat liver allograft tolerance to rejection. AB - BACKGROUND: Previous studies showed that liver transplant rejection in the Piebald Virol Glaxo (PVG)-to-Lewis combination was associated with more intragraft interleukin (IL)-4 mRNA expression than in spontaneously tolerant grafts in the PVG-to-Dark Agouti (DA) combination. There was also immunoglobulin (Ig) G1 antibody deposition, suggesting an IL-4-induced IgG class switch in rejection. The aim of this study was to investigate whether IL-4 treatment converts PVG-->DA liver transplant tolerance to rejection. METHODS: DA (RT1a) rats were recipients of orthotopic PVG (RT1c) liver transplants and DA liver transplants were syngeneic controls. Supernatant from IL-4-transfected Chinese hamster ovary cells (0.5 mL, 30,000 U) or from untransfected cells was injected intraperitoneally on days 3 through 7. Samples were taken for immunohistochemical staining of frozen tissue sections to analyze cellular infiltrate and antibody deposition. RESULTS: IL-4 treatment significantly reduced survival of liver allografts from greater than 100 days in untreated animals to 9 days (P=0.004). Pathologic analysis of IL-4-treated animals showed that death was caused by liver transplant rejection, with a heavy infiltrate of mononuclear cells, disruption of portal tract areas, and infarction. Immunohistochemistry revealed an extensive infiltrate of T cells, CD25-expressing cells, and B cells that was similar to the level in PVG--> Lewis liver allograft recipients that reject the liver. There was also a more extensive monocyte-macrophage infiltrate and more major histocompatibility complex class II expression in IL-4-treated animals compared with untreated animals. There was moderate increase of IgM, little IgG1, and no IgE or IgG2a antibody deposition. CONCLUSIONS: IL-4, a T-helper type 2 cytokine that has previously been shown to inhibit delayed-type hypersensitivity responses such as rejection, was found to promote rejection of liver allografts. There was only slight evidence of a graft-specific antibody response, showing that IL-4 induces liver allograft rejection in association with some, but not all, of the changes accompanying rejection in the PVG-->Lewis strain combination. PMID- 15880054 TI - Prevention of acute vascular rejection by a functionally blocking anti-C5 monoclonal antibody combined with cyclosporine. AB - BACKGROUND: Inhibition of the complement cascade at C5 prevents formation of pro inflammatory molecules C5a and C5b-9, which play a key role in allograft rejection. The present study was undertaken to determine whether blocking terminal complement with anti-C5 monoclonal antibody (mAb) alone or combined with cyclosporine (CsA) would prevent acute vascular rejection (AVR) in a mouse cardiac allograft model. METHODS: C3H mouse hearts were transplanted into BALB/c mice and randomized into five groups with the following treatments: (1) no treatment; (2) CsA alone; (3) control mAb; (4) anti-C5 mAb alone; and (5) anti-C5 mAb and CsA. RESULTS: Allografts in untreated or control mAb-treated recipients were rapidly rejected at 8.0+/-0.6 and 8.2+/-0.8 days, respectively. These grafts exhibited typical AVR, characterized by vasculitis, hemorrhage, and thrombosis. A high level of complement activity was also demonstrated in these animals. High dose CsA was not able to inhibit complement activation or AVR, and grafts were rejected in 15.5+/-1.1 days. Anti-C5 monotherapy completely inhibited complement activation and attenuated AVR, but grafts were rejected in 8.3+/-0.5 days by acute cellular rejection. In contrast, a combination of anti-C5 mAb and CsA successfully achieved indefinite graft survival (>100 days). This combined therapy completely inhibited terminal complement activation and prevented both humoral- and cellular-mediated rejection. CONCLUSIONS: Combination therapy of anti-C5 mAb and CsA achieves indefinite graft survival in a mouse cardiac allograft model. These data suggest that inhibition of terminal complement using anti-C5 mAb may be an effective therapeutic adjunct to prevent AVR in clinical transplantation. PMID- 15880055 TI - Comparison of hepatic coagulant, fibrinolytic, and anticoagulant functions between Banna Minipig Inbred line and humans. AB - BACKGROUND: As an ideal candidate for xenotransplantation, the compatibility of physiological porcine organs with those of humans is an essential premise. In this study, we analyzed hepatic coagulant, fibrinolytic, and anticoagulant functions between Banna Minipig Inbreds (BMIs) and humans to evaluate such hepatic compatibility. METHODS: BMI factors II, V, VII, X, and XII were added to the corresponding factor-deficient human plasma to determine prothrombin times (PT) and activated partial thromboplastin times (APTT). Human tissue plasminogen activator (t-PA) was added to both BMI and human plasma to determine plasmin activity. The antithrombin-III (AT-III) activity of plasma was analyzed with the STA-Stago autoanalyzer using an AT-III assay kit. RESULTS: Both PT and APTT were reduced but within normal parameters when BMI factors II, V, VII, X, and XII were added to the corresponding factor-deficient human plasma. The activities of BMI coagulation factors II, V, VII, X, and XII were 3.2, 3.7, 4.7, 2.9, and 4.5 times those of humans, respectively. The activity of plasmin was significantly higher in BMI plasma than in humans when human t-PA was added to both. The normal range of human AT-III activity was 90-108% while BMI AT-III was 124.50 +/- 2.38%. CONCLUSIONS: The activities of coagulation factors and AT-III were higher in BMIs than in humans. BMI coagulation factors XII, VII, and X trigger human intrinsic, extrinsic, and common pathways, respectively, which functioned normally. In addition, BMI plasminogen could be activated by human t-PA. PMID- 15880056 TI - Natural history of hepatitis B and C in renal allograft recipients. AB - BACKGROUND: In renal allograft recipients, most cases of liver dysfunction are caused by hepatitis B virus and hepatitis C virus (HCV). The natural history of hepatitis C and B was studied in 286 renal allograft recipients who received a kidney allograft between 1972 and 1989 when tests for anti-HCV became available. METHODS: In all patients, hepatitis B (HB) surface (s) antigen (Ag) was tested before and anti-HCV (by enzyme-linked immunosorbent assay II) after transplantation. RESULTS: At enrollment in 1989 (5.5+/-4 years after transplantation), 209 patients were anti-HCV positive (C+), 42 patients were HBsAg-positive (B+), and 35 patients were both B+ and C+ (C+B+). One hundred four patients were receiving azathioprine (AZA) and 182 were on cyclosporine A (CsA). Since transplantation, the median follow-up was 18 years in AZA-treated and 13 years in CsA-treated patients. Liver biopsy showed chronic hepatitis in 73 patients, cirrhosis in 20 patients, and fibrosing cholestatic hepatitis in 2 patients. In 34 patients, liver biopsy was repeated, and progression of fibrosis was observed in 24 patients. The 12-year patient survival rate was similar in B+, C+, and B+C+ patients (67%, 78%, and 71%, respectively; P=not significant). Liver related death was the first cause of death in B+ and B+C+ infected patients (58% and 72%, respectively), whereas cardiovascular disease was the leading cause of death in C+ patients (40%). Multivariate analysis showed that older age (>40 years) (relative risk [RR], 2.8), B+ status (RR, 2.36), and C+ status (RR, 1.65) were independently associated with a worse patient survival. CONCLUSIONS: In the long term, B+ patients had a higher risk of death related to liver disease than C+ patients, and co-infection did not worsen patient survival. PMID- 15880057 TI - A technique for retroperitoneal pancreas transplantation with portal-enteric drainage. AB - BACKGROUND: Pancreas transplantation (PTx) with portal-enteric drainage (PED) has been associated with difficulties in respect to arterial anastomosis and graft accessibility for percutaneous biopsy. We describe a new technique that circumvents these difficulties. METHODS: Between April 2001 and April 2004, a total of 113 recipients were scheduled for PTx with PED. The superior mesenteric vein was approached from the right retroperitoneal aspect instead of from the anterior transmesenteric route. The pancreas graft was eventually placed in the right retroperitoneal space, being covered by the ascending colon and its mesentery. RESULTS: One hundred ten (97.3%) PTx were performed as planned. Systemic venous effluent was preferred in three patients because of incidental diagnosis of liver cirrhosis during surgery (n=1) and severe obesity (body mass index>35 kg/m2) (n=2). The Y iliac artery graft was kept as short as possible, and arterial anastomosis was always performed with ease. After a mean follow-up period of 21.2+/-19.9 months, the relaparotomy rate was 13.6%. No patient died after repeat surgery, and none required multiple relaparotomies. Overall, 10 grafts were lost because of acute rejection (n=3), chronic rejection (n=2), venous thrombosis (n=2), recipient death (n=2), and late (6-month) arterial thrombosis (n=1). One-year patient and graft survival were 98.1% and 90.7%, respectively. CONCLUSIONS: Our data confirm that PTx with PED is not associated with an increased risk. The technique described has distinctive technical advantages and should be included in the repertoire of PTx. PMID- 15880058 TI - Ethical and legal issues in non-heart-beating organ donation. AB - Procurement of kidneys and livers from non-heart-beating donors (NHBD) raises ethical and legal issues that need to be considered before wider use of these donors is undertaken. Although NHBDs were used in kidney transplantation as early as the 1960s, retrieval of these organs is not universally accepted today. From a medical point of view, these organs were considered "marginal" because the majority showed delayed or impaired function early after implantation. Legal problems relate to determination of death on cardiopulmonary criteria, the issue of valid consent, and the use of preservation measures. Among ethical issues involved are observance of the dead-donor rule, decisions with respect to resuscitation and withdrawal of life-sustaining treatment, respect for the dying patient and the dead body, and proper guidance of the family. In The Netherlands NHB donation was pioneered by the Maastricht Centre as early as 1981. Today, all seven transplant centers procure and transplant these organs, and NHBDs have become an important source of transplantable kidneys and livers. Recent legislation in The Netherlands also supports NHB donation by allowing the use of organ-preserving measures, even in the absence of family consent. As a result, one of every three kidneys transplanted in The Netherlands in 2004 derives from a NHBD. This article explores Dutch NHBD practice, protocols, and results and compares these data internationally. PMID- 15880059 TI - Split-liver transplantation with pediatric donors: a multicenter experience. AB - BACKGROUND: Outcomes of split-liver transplantation (SLT) with pediatric donors have never been specifically reported. METHODS: A prospective multicenter study on SLT using donors younger than 15 years was conducted. Thirty-nine split-liver procedures generating a left lateral segment (LLS) and an extended right graft (ERG) were performed. In three cases, no recipient was found for ERG. In all but one case, the celiac trunk was maintained with LLS. Data were available for 67 grafts (90% of the total): 38 LLSs and 9 ERGs transplanted into 46 children and 20 ERGs transplanted into 20 adults. Sixty-two (93%) grafts were used for primary transplants and five (7%) for retransplantation. SLT were performed with 15 donors 10 years of age and less and with 24 between 11 and 15 years. RESULTS: Median follow-up was 24 months. Two-year patient and graft survival were 87% and 82%. Patient and graft survivals were not significantly different between pediatric and adult recipients, between recipients from donors 10 years of age and less and those between 11 and 15 years, and between recipients of LLS and ERG. Arterial complications occurred in 6% of cases (8% in the < or = 10 year donors group, 5% in the 11-15 year donors group). The incidence of other complications was similar between groups. CONCLUSIONS: SLT with pediatric donors, even younger than 10 years, provided results comparable with those achievable using adult donors. The similar incidence of arterial complications among patients receiving LLS or ERG suggests that maintenance of the celiac trunk with LLS is not detrimental for right-sided grafts. PMID- 15880060 TI - Evaluation of pretransplant immunologic status in kidney-transplant recipients by panel reactive antibody and soluble CD30 determinations. AB - OBJECTIVES: To retrospectively compare the accuracy of pretransplant panel of reactivity antibodies (PRA) and serum level of soluble CD30 (sCD30) in predicting early (<6 months) acute rejection (AR) in living-donor and deceased-donor kidney transplant (KT) patients. METHODS: Pretransplant sera of 24 KT recipients were retrospectively tested for sCD30 and compared with PRA. Inclusion criteria were de novo graft patients on calcineurin-inhibitor-based immunosuppression, minimum follow-up of 1 year, alive with a functioning graft, and stable renal function over the last 12 months. Objective measures were incidence of biopsy-proven AR (BPAR) within 6 months of KT and sCD30 and PRA diagnostic indexes. The relative risk (RR) of BPAR for each test was also obtained. RESULTS: Fourteen (58.3%) patients presented at least one episode of BPAR within 6 months of KT. All rejection episodes were responsive to steroid treatment. PRA was positive in six (25%) patients, and four (66.7%) of them presented at least one episode of BPAR. sCD30 tested positive in nine (37.5%) patients, and all these later presented at least one episode of BPAR. sCD30 and PRA diagnostic indexes in predicting early (< 6months) BPAR were sensitivity 64.2% versus 28.5%; specificity 100% versus 80%; accuracy 79.1% versus 50%; positive predictive value 100% versus 66.6%; and negative predictive value 66.6% versus 44.4%. The RR of early AR was 1.4 in PRA positive patients and extremely higher in the sCD30-positive group. CONCLUSIONS: Pretransplant sCD30 is a more accurate predictor of AR when compared with PRA. These results support its use in the pretransplant work-up of kidney-graft recipients. PMID- 15880061 TI - Tolerance: is it worth the risk? AB - BACKGROUND: The success of orthotopic liver transplantation (OLT) has been limited by the adverse effects of immunosuppression. The purpose of this study was to determine the safety and feasibility of withdrawing immunosuppression in OLT recipients to achieve tolerance. METHODS: Eighteen adult OLT recipients in our steroid-free protocol without rejection were selected for this protocol. All patients chosen for this trial were on tacrolimus monotherapy with normal liver function tests (LFTs). Tacrolimus was weaned as long as LFTs remained stable. Weaning was halted for elevations of liver enzymes and tacrolimus was increased to the last dosage at which the patients had normal LFTs. Rejection was treated by increasing tacrolimus to levels of 10-15 ng/ml. Mycophenolate mofetil (MMF) or sirolimus was added if there was severe rejection by biopsy. Steroids were used if there was no improvement. RESULTS: One patient has been weaned off immunosuppression. Three additional patients were weaned completely off but had tacrolimus resumed because of mild elevations in LFTs. Eleven of 18 (61%) patients had rejection. Two patients required steroid therapy and one required rabbit antithymocyte globulin in addition to MMF and steroids. One of the patients with rejection developed diabetes and one patient had renal failure, which subsequently resolved. One patient died following a stroke. CONCLUSIONS: Clinical tolerance can be achieved in a minority of patients, even when being maintained on minimum immunosuppression. The potential benefit of achieving tolerance must be weighed against the risks of rejection therapy in patients doing well on low-dose immunosuppression. PMID- 15880062 TI - Renal dysfunction as a risk factor for mortality and cardiovascular disease in renal transplantation: experience from the Assessment of Lescol in Renal Transplantation trial. AB - BACKGROUND: Renal-transplant recipients have shortened life expectancy primarily because of premature cardiovascular disease. Traditional and nontraditional risk factors for cardiovascular disease are prevalent in renal patients. In renal transplant recipients, immunosuppressive therapy can be nephrotoxic and aggravate cardiovascular disease risk factors. Renal dysfunction has been established as a risk factor for cardiovascular disease and mortality in different populations. We evaluated the effects of baseline renal-transplant function on mortality and cardiovascular and renal endpoints in 1,052 placebo-treated patients of the Assessment of Lescol in Renal Transplantation trial. METHODS: All renal transplant recipients were on cyclosporine-based immunosuppressive therapy. Follow-up was 5 to 6 years, and endpoints included cardiac death, noncardiovascular death, all-cause mortality, major adverse cardiac event (MACE), stroke, nonfatal myocardial infarction, and graft loss. RESULTS: Baseline serum creatinine was strongly and independently associated with increased cardiac, noncardiovascular, and all-cause mortality, as well as MACE and graft loss. Serum creatinine was not a risk factor for stroke or nonfatal myocardial infarction. CONCLUSIONS: Elevated baseline serum creatinine in renal-transplant recipients is a strong and independent risk factor for all-cause, noncardiovascular and cardiac mortality, MACE, and graft loss. PMID- 15880063 TI - Pharmacokinetic of cyclosporine microemulsion in pediatric kidney recipients receiving A quadruple immunosuppressive regimen: the value of C2 blood levels. AB - BACKGROUND: The management of cyclosporine therapy in pediatric kidney-transplant recipients is largely based on single center's experience rather than on a univocal pharmacokinetic approach based on therapeutic drug monitoring. A prospective multicenter trial was designed to address the question whether C2 blood level monitoring of cyclosporine microemulsion therapy is feasible in the pediatric setting. METHODS: Sixty-four pediatric kidney-transplant recipients receiving a triple immunosuppressive regimen based on cyclosporine microemulsion had their cyclosporine dose adjusted to the same protocol-defined C2 targets from the time of the transplant until 2 years posttransplant. The interim analyses after 1 year of enrollment is presented in this study. RESULTS: One-year patient and graft survival were 100% and 94.8%, respectively. One-year rejection rate was 15%. C2 management of cyclosporine did not affect graft function: 1-year serum creatinine and glomerular filtration rate were 1.3+/-1 mg/mL and 71.2+/-20 mL/min/1.73 m2, respectively. C2 was the best single-point predictor of the area under the concentration curve throughout the entire follow-up, with a mean coefficient of correlation of 0.97+/-0.01. CONCLUSIONS: C2 management of cyclosporine microemulsion therapy is effective and safe in pediatric kidney transplant recipients given a combined immunosuppressive treatment. PMID- 15880064 TI - Living-unrelated donors yield higher graft survival rates than parental donors. AB - BACKGROUND: Living-unrelated donors (LURD) have been shown to yield kidney graft survival rates equivalent to that of related donors. Here, we show that in three diseases, LURDs supply grafts that survive at a higher rate than parental or offspring donors. METHODS: We analyzed 111,643 first adult kidney transplants from the United Network for Organ Sharing database performed between 1991 and 2003 by Kaplan-Meier curves, log-rank tests, and Cox proportional hazard tests. RESULTS: Five-year kidney graft survival rates in patients receiving grafts from LURD, parental, and offspring donors were 75.4%, 74.3%, and 75.7% among patients with typical original diseases excluding three diseases. In focal glomerulosclerosis (FGS), the corresponding results were 87.4%, 70.1%, and 81.6%. In polycystic kidney disease (PC) rates were 86.4%, 78.6%, and 85.3%, and in diabetes mellitus (type 1 DM) rates were 75.9%, 69.8%, and 70.5%. Parental donors yielded significantly lower graft survival rates than LURDs in all three diseases. Although statistically significant differences were lost in the Cox tests, functional graft survival showed significantly lower graft survival rates from parental donors compared with LURDs, suggesting that donor age was a significant factor. CONCLUSIONS: If living donors are considered for kidney transplantation into patients with FGS, PC, or type 1 DM, grafts from LURD are preferred over parental and offspring donors. Although genetic susceptibility of parental and offspring donor grafts may be a factor, the simple donor age factor probably results in lower survival of parental donor grafts. Lower survival of offspring donor grafts may suggest genetic susceptibility because such donors were, in general, younger than in LURD grafts. PMID- 15880065 TI - Early and midterm risk of coronary allograft vasculopathy in patients bridged to orthotopic heart transplantation with ventricular assist devices. AB - BACKGROUND: Coronary transplant vasculopathy (CAV) has been associated with both immunologic and nonimmunologic factors. The impact of preoperative ventricular assist device (VAD) support on the development of CAV has not been studied. To examine this, we obtained posttransplant coronary angiograms from a group of patients bridged with VAD and compared them to post transplant coronary angiograms of a non-VAD cohort. METHODS: Adult patients undergoing orthotopic heart transplant between 1996-2000 were retrospectively studied and divided into VAD and non-VAD patients. Coronary angiograms were retrospectively reviewed and severity of coronary vasculopathy was categorized as trivial, mild, moderate, or severe. Other variables studied included recipient and donor demographics, cytotoxic panel reactive antibodies (PRA) against T-cell targets and flow cytometric crossmatching against donor T lymphocytes. RESULTS: There was no significant difference between groups regarding demographics. However, VAD patients had a sixfold greater chance of having a T-cell PRA >10% at the time of transplant (p < 0.05), and a fourfold greater chance of having a positive cross match when compared to non-VAD patients (p < 0.05). There was no significant difference in the degree of CAV between groups. Normal coronary anatomy was present in 76% of VAD patients and 64% of non-VAD patients (p = 0.37). These results were similar at 2- and 3-year follow-up (76 vs. 74% and 80 vs. 62%, respectively). CONCLUSION: Preoperative VAD use is associated with increased sensitization; however, these patients develop CAV at the same rate as those not bridged with a VAD. PMID- 15880066 TI - Augmented Induction of CD4+CD25+ Treg using monoclonal antibodies to CD200R. AB - BACKGROUND: CD200 is a transmembrane protein delivering immunoregulatory signals after engagement of CD200R. A family of CD200Rs exist (CD200R1-4) with different tissue expression and functional activity. In the presence of anti-CD200R2/3 monoclonal antibodies (mAbs), bone-marrow cells cultured in the presence of (interleukin [IL]-4+granulocyte-macrophage colony-stimulating factor) differentiate into dendritic cells (DCs), which induce CD4+CD25+ Treg. The effect of these mAbs on Treg induced in anti-CD3 activated thymocyte cultures is unknown. METHODS: BL/6 bone-marrow cells were cultured with GMC-SF and IL-4 in the presence/absence of anti-CD200Rs to generate DCs that induced Treg in C3H lymph-node T cells. Treg were also induced in anti-CD3/CD28-activated C3H thymocytes. Treg activity was assayed by (1) suppression of mixed leukocyte culture (MLC) in cultures using C3H stimulator spleen cells and BL/6 stimulator cells, and (2) by expression of the transcription factor, Foxp3. RESULTS: Addition of anti-CD200R2/3 mAbs (but not anti-CD200R1) to bone-marrow cultures led to generation of DCs that induced a CD4+CD25+ (Treg) population inhibiting MLCs (C3H cells stimulated with C57BL/6 cells) in a cytotoxic T-lymphocyte associated antigen (CTLA)-4 and transforming growth factor (TGF)-beta-dependent manner. Anti-CD200R2, but not anti-R1/R3, augmented induction of Foxp3-expressing Treg from anti-CD3/CD28 activated thymocytes. Suppression in MLCs by anti-CD200R1 mAbs was dependent on IL-10 and TGF-beta. CONCLUSIONS: Unlike anti-CD200R1, anti CD200R2 both promotes development of DCs with capacity to induce Treg and directly augments thymocyte production of Treg. PMID- 15880067 TI - Calcineurin inhibitor- and steroid-free immunosuppression in pancreas-kidney and solitary pancreas transplantation. AB - BACKGROUND: Calcineurin inhibitors and steroids are standard immunosuppressants in solid organ transplantation, but (long-term) side effects are harmful to the recipient and the graft. The authors present a novel strategy for posttransplant immunosuppression that combines a depleting antibody with an antimetabolite, avoiding calcineurin inhibitors and steroids. METHODS: In a prospective, nonrandomized, observational cohort study, 75 pancreas-kidney and solitary pancreas recipients received alemtuzumab (4 doses for induction and up to 12 doses within the first year) and mycophenolate mofetil (> or = 2 g/day) for induction and maintenance therapy. Minimum follow-up was 6 months. We compared the results with a historical group of 266 consecutive pancreas recipients on Thymoglobulin (induction) and tacrolimus (maintenance). RESULTS: Differences in patient and graft survival rates between the study and control groups at 6 months were not statistically significant. However, the incidence of a first reversible rejection episode was significantly higher for simultaneous pancreas-kidney recipients in the study (vs. control) group. We noted a trend toward higher modification of renal disease levels at 6 months posttransplant in the study (vs. control) groups. CONCLUSIONS: The combination of alemtuzumab and mycophenolate mofetil was associated with an acceptable rejection rate, a good safety profile, and good (graft and native) kidney function; it eliminated undesired calcineurin inhibitor- and steroid-related side effects. Longer follow-up is warranted before expanded application can be recommended. PMID- 15880068 TI - Direct visualization of cortical peritubular capillary of transplanted human kidney with reperfusion injury using a magnifying endoscopy. AB - BACKGROUND: We developed a direct imaging system of renal microcirculation by a magnifying-endoscopy that enables visualization of the movement of erythrocyte in glomerular and cortical peritubular capillary (CPC). We investigated the microcirculation of CPC in the early phase of both living- and cadaveric-donor transplant kidneys. METHODS: Erythrocyte velocity in CPC were monitored and measured in 20 renal transplants at 20, 60, 90, and 120 minutes after reperfusion. The kidney grafts came from 11 living donors and 9 non-heart-beating cadaveric donors. RESULTS: In living-donor transplants, erythrocyte velocity in CPC at 20 minutes after revascularization declined to one third of baseline value just before nephrectomy and recovered to the prenephrectomy value 120 minutes after reperfusion. In contrast, it continued to be disturbed for 90 minutes in cadaveric-donor transplants. Erythrocyte velocity in CPC more significantly deteriorated in cadaveric transplants than in those of living transplants at 20 through 60 minutes after the revascularization. In living-donor transplants, erythrocyte velocity did not correlate with donor age, both warm (WIT) and cold ischemic time (CIT), time to the initial urination, and best creatine clearance. In the cadaveric transplants, ischemic time, both WIT and CIT, did not correlate with the erythrocyte velocity. However, donor age, duration of acute tubular necrosis, and best creatine clearance after transplantation significantly correlated with the erythrocyte velocity. CONCLUSION: The measurement of erythrocyte velocity in CPC is a reliable method for predicting the recovery of renal function and reserved renal function of kidney allografts undergoing prolonged ischemia. PMID- 15880069 TI - Non-heart-beating donor kidneys in the Netherlands: allocation and outcome of transplantation. AB - BACKGROUND: Since February 1, 2001, kidneys from both heart-beating (HB) and non heart-beating (NHB) donors in The Netherlands have been indiscriminately allocated through the standard renal-allocation system. METHODS: Renal function and allograft-survival rate for kidneys from NHB and HB donors were compared at 3 and 12 months. RESULTS: The outcomes of 276 renal transplants, 176 from HB donors and 100 from NHB III donors, allocated through the standard renal allocation system, Eurotransplant Kidney Allocation System, and performed between February 1, 2001 and March 1, 2002 were compared. Three months after transplantation, graft survival was 93.7% for HB kidneys and 85.0% for NHB kidneys (P<0.05). At 12 months, graft survival was 92.0% and 83.0%, respectively (P<0.03). Serum creatinine levels in the two groups were comparable at both 3 and 12 months. Multivariate analysis identified previous kidney transplantation (relative risk [RR] 3.33; P<0.005), donor creatinine (RR 1.01; P<0.005), and NHB (RR 2.38; P<0.05) as independent risk factors for transplant failure within 12 months. In multivariate analysis of NHB data, a warm ischemia time (WIT) of 30 minutes or longer (P<0.005; RR 6.16, 95% confidence interval 2.11-18.00) was associated with early graft failure. No difference in 12-month graft survival was seen between HB and NHB kidneys after excluding the kidneys that failed in the first 3 months. CONCLUSION: Early graft failure was significantly more likely in recipients of kidneys from NHB donors. A prolonged WIT was strongly associated with this failure. Standard allocation procedures do not have a negative effect on outcome, and there is no reason to allocate NHB kidneys differently from HB kidneys. PMID- 15880070 TI - Logistics and transplant coordination activity in the GRAGIL Swiss-French multicenter network of islet transplantation. AB - BACKGROUND: Since the Edmonton trial in 2000, increasing numbers of transplant centers have been implementing islet transplantation programs. Some institutions have elected to associate in multicenter networks, such as the Swiss-French GRAGIL (Groupe Rhin-Rhone-Alpes-Geneve pour la Transplantation d'Ilots de Langerhans) consortium. METHODS: All pancreata offers to the University of Geneva Cell Isolation and Transplantation Center from within the network in 2002 and 2003 were reviewed. Islet preparations were attributed to the most suitable recipient on a centrally managed waiting list. All shipments were performed by ambulance in less than 5 hr. RESULTS: Over the period of study, 260 pancreata were offered, from a total of 1,304 cadaveric donors in the four allocation regions (20%). Fifty-two patients were on the waiting list at any time during this 2-year period. The percentage of organs offered varied in the range of 0.5% to 42%, depending on region of origin, with a correlation with number of patients on the waiting list in each region. Of these, 104 (40%) were accepted for processing. Ninety-two pancreata were actually processed, resulting in 42 islet preparations being transplanted. The number of international equivalents of transplanted preparations was 378,500+/-16,000 versus 165,400+/-15,400 (P<0.0001) for nontransplanted preparations. Total cold ischemia time was 6+/-0.3 hr for transplanted preparations versus 6.7+/-0.4 hr for nontransplanted preparations (not significant). CONCLUSIONS.: A high rate of pancreas offers, successful isolation, and islet transplantation can be achieved in multicenter networks such as GRAGIL. Such an approach can expand both the donor pool and the recipient population. PMID- 15880071 TI - Pediatric transplantation: the Hamburg experience. AB - BACKGROUND: Since starting our program in 1989, 455 pediatric orthotopic liver transplantations have been performed using all techniques. In April 2001, we experienced our last in-hospital death of a pediatric liver-transplant recipient. Since then, all our liver-transplant children (n=170) were able to be discharged from the hospital. The aim of this study is to analyze the actual status of pediatric liver transplantation at the University of Hamburg and to find future perspectives to improve the results after pediatric liver transplantation. METHODS: From May 4, 2001 until September 8, 2004, 22 (13%) whole organs, 18 (11%) reduced-size organs, 79 (47%) split organs, and 51 (30%) organs from living donors were transplanted into 142 patients. One hundred forty-one were primary liver transplants, 25 retransplants, 3 third, and 1 fourth liver transplants. Of the 170 orthotopic liver transplantations (OLT), 31 (18%) were highly urgent (United Network of Organ Sharing [UNOS] I). RESULTS: After 170 consecutive pediatric liver transplants, no patients died during the hospital course (100% patient survival<3 months), but overall, 5 (2.9%) recipients died during further follow-up. The 3-month and actual graft survival rates are 93% and 85%, respectively. Twenty (11.8%) children had to undergo retransplantation. However, patient survival was not sustained by longer graft survival. Analyzing our series, we see that graft survival after reduced-size liver transplantation showed a significantly lower rate versus living-donor liver transplantation. CONCLUSION: The learning curve in pediatric liver transplantation has reached a turning point where immediate patient survival is considered the rule. The challenge is to increase graft survival to the same level. The long-term management of the transplant patients, with the aim of avoiding late graft loss and achieving excellent quality of life, will become the center of the debate. PMID- 15880072 TI - The International Registry on Hand and Composite Tissue Transplantation. AB - BACKGROUND: Since May 2002 all groups performing hand transplantations have supplied detailed information to the International Registry on Hand and Composite Tissue Transplantation. This inaugural report provides a review of all hand transplants performed to date. METHODS: Between September 1998 and September 2004, 18 male patients underwent 24 hand/forearm/digit transplantations (11 monolateral and 4 bilateral hand transplantations, 2 bilateral forearm transplantations, and 1 thumb transplantation). The level of amputation was mostly at the distal forearm or wrist. The average age of the patient was 32 years. Time since hand loss ranged from 2 months to 22 years. Immunosuppressive therapy included tacrolimus, mycophenolate mofetil, rapamycin, and steroids; polyclonal or monoclonal antibodies were used for induction. Topical immunosuppression was administered in some patients. Follow-up period ranged from 17 to 70 months. RESULTS: Patient survival was 100%. Graft survival was 100% at 1 and 2 years. Two cases of graft failure at a later date were caused by severe inflammation and progressive rejection in a noncompliant patient. Acute rejection episodes occurred in 12 patients within the first year. Rejection was reversible in all compliant patients. Side effects included opportunistic infections and metabolic complications. No life-threatening complications or malignancies were reported. All patients had achieved protective sensation, and 17 patients also achieved discriminative sensation. Extrinsic and intrinsic muscle recovery enabled patients to perform most daily activities. CONCLUSIONS: Despite the enormous antigen load associated with composite tissue allograft, hand transplantation became a clinical reality with immunosuppression comparable to transplantation of solid organs. PMID- 15880073 TI - High CXCL10 expression in rejected kidneys and predictive role of pretransplant serum CXCL10 for acute rejection and chronic allograft nephropathy. AB - BACKGROUND: Several experimental models have shown that CXCL10 is required for initiation and development of graft failure caused by both acute and chronic rejection. METHODS: CXCL10 expression and distribution was investigated in tissue specimens obtained from 22 patients suffering from acute rejection (AR) or chronic allograft nephropathy (CAN) by using in situ hybridization. Furthermore, pretransplantation sera of 316 cadaveric kidney-graft recipients were tested retrospectively for serum CXCL10 levels by a quantitative sandwich immunoassay. RESULTS: Bioptic specimens obtained from patients with CAN were characterized by wide CXCL10 expression not only at level of infiltrating inflammatory cells but also of vascular, tubular, and glomerular structures. In addition, assessment of pretransplant serum CXCL10 levels in 316 graft recipients and stratification of patients in three groups according to serum CXCL10 levels (<100 pg/mL, n=163; 100 150 pg/mL, n=69; >150 pg/mL, n=84) showed highly significant differences in 5 year survival rates for the two extreme groups (95.7% vs. 79.7%, P=0.0002). Accordingly, patients who developed severe, early AR (277.14+/-65.08 p=0.004) and those who developed CAN also showed increased pretransplant serum CXCL10 levels (193.2+/-36.9, P=0.03). Multivariate analysis demonstrated that among the analyzed variables, CXCL10 (relative risk [RR] 2.801) and delayed graft function (RR 3.728) had the highest predictive power of graft loss. CONCLUSIONS: These results suggest that pretransplant serum CXCL10 levels greater than 150 pg/mL confer an increased risk of early, severe, AR and subsequent CAN, finally resulting in renal-allograft failure. This finding might be used for the individualization of immunosuppressive therapies. PMID- 15880074 TI - The beneficial effects of recipient-derived vascular endothelial growth factor on graft survival after kidney transplantation. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is crucial for preservation of microvasculature and contributes to cytoprotection of the graft after kidney transplantation. METHODS: The authors investigated the influence of VEGF single nucleotide polymorphism (SNP) on graft survival after renal transplantation. The SNP at positions -2578, -1154, and, -7 were analyzed in 306 donors and 387 recipients of renal transplants. RESULTS: The authors observed no effect of those recipient or donor SNP on acute rejection. However, graft survival was associated with recipient SNP at position -2578 C/A. Recipients with a genetic basis for high production of VEGF had significantly better graft survival compared with recipients with low production of VEGF. Homozygotes for the A allele (low producers of VEGF) had worse graft survival compared with high producers, the heterozygotes and homozygotes for C allele (P=0.03). Multivariate analysis in which the effects of donor age, recipient race, cold ischemia time, donor origin, and number of human leukocyte antigen mismatches were included showed that the status of noncarriers of -2578 C allele of recipients was an independent factor for graft failure (odds ratio, 1.8; 95% confidence interval, 1.0-3.0; P=0.03). CONCLUSIONS: The authors conclude that homozygote recipients for the -2578 A allele, the low producers of VEGF, are more vulnerable to tissue injury, resulting in worse graft survival. PMID- 15880075 TI - Livers from non-heart-beating donors tolerate short periods of warm ischemia. AB - BACKGROUND: In contrast with kidneys, transplantation of livers originating from non-heart-beating donors remains rare, mainly because warm ischemia causes a higher rate of potentially lethal primary graft nonfunction. Little is known on the tolerance of liver grafts to warm ischemia. No techniques are available to assess the viability of ischemic livers before implantation. Therefore, experimental models are needed to address these questions before non-heart beating liver transplantation can be more widely applied. This study aims to develop a reproducible large animal model of liver transplantation using non heart-beating donors and, in this model, to define the tolerance of the liver to warm ischemia. METHODS: Pigs weighing 25to 30 kg are used. In donors, cardiac arrest is caused by ventricular fibrillation. After increasing lengths of warm ischemia (0, 15, 30, 45, and 60 min), the liver is flushed in situ with 4 degrees C histidine tryptophan ketoglutarate preservation solution and procured. The liver is transplanted after a 4-hour cold storage period. RESULTS: Control livers (no warm ischemia) and livers exposed to 15 minutes of warm ischemia function normally after transplantation, whereas all livers submitted to 60 minutes of warm ischemia display primary nonfunction and cause recipient death. Graft function and survival are occasionally observed after 30 and 45 minutes of warm ischemia. CONCLUSIONS: A reproducible model of non-heart-beating liver transplantation is described. We found that the liver tolerates 15 minutes of warm ischemia. This preclinical model is a valid tool to develop techniques to predict the quality of ischemic livers before implantation and to design interventional strategies to improve the tolerance of the liver to warm ischemia. PMID- 15880076 TI - Apoptosis in kidney and pancreas allograft biopsies. AB - BACKGROUND: Apoptosis is a particular form of cell death involved in the elimination of somatic cells. In this study, the occurrence of apoptotic cells in kidney and pancreas allograft biopsies was analyzed and correlated with the number of infiltrating macrophages and lymphocytes and granzyme B expression. METHODS: Kidney and pancreas biopsies from patients submitted to simultaneous pancreas-kidney transplantation were classified into three groups: acute rejection, chronic rejection, and transplant cases without evidence of rejection. Formalin-fixed paraffin biopsies were used to identify apoptosis by the terminal deoxynucleotidyl transferase [TdT]-mediated dUTP nick end labeling (TUNEL) method. RESULTS: In normal kidney, only few apoptotic cells were observed. In contrast, in kidney-allograft biopsies, the TUNEL signal was detected in the nuclei of tubular epithelial cells and also in mononuclear cells scattered in the interstitium. In pancreas biopsies, numerous apoptotic cells were detected in acinar cells, in ducts, and occasionally in islets. The number of apoptotic cells in acute pancreas rejection was significantly higher compared with acute rejection of kidney grafts (50+/-14 vs. 21+/-4 cells/mm2; P<0.05). In kidney biopsies, there was a positive correlation between apoptosis and macrophages (r=0.51; P<0.005), and apoptosis versus T lymphocytes (r=0.45; P<0.05). In pancreas biopsies, the number of apoptotic cells correlated only with the number of macrophages (r=0.41; P<0.05). CONCLUSIONS: Apoptosis occurs in kidney and pancreas allograft biopsies, markedly in acute rejection in pancreas biopsies. Although apoptosis may reflect a mechanism of down-regulation of the allograft immune response by eliminating infiltrating cells, the elimination of graft cells may result in graft damage, particularly in pancreas transplantation. PMID- 15880077 TI - Laparoscopic versus open living-donor nephrectomy: experiences from a prospective, randomized, single-center study focusing on donor safety. AB - BACKGROUND: Very few randomized studies on laparoscopic (L) versus open (O) living-donor nephrectomy (LDN) have been presented. The largest randomized series reported so far included 80 donors. In 2000, an Australian safety group concluded that the evidence base for L-LDN is inadequate to make recommendations regarding safety and efficacy. METHODS: With this background, at our single national center, 122 donors were randomized to left-sided L-LDN (n=63) or O-LDN (n=59), from February 2001 to May 2004. This article summarizes our experiences, in particular regarding complications and safety. RESULTS: There were significant differences in favor of O-LDN regarding operative time, warm ischemia time, and vessel lengths, whereas the analgesic requirements and pain data were significantly in favor of the laparoscopic procedure. In the L-LDN group, there were five major postoperative complications resulting in reoperations (8%), including two intestinal perforations. No major complications occurred in the O LDN group. CONCLUSIONS: These results from our randomized study do suggest that conventional O-LDN is a very secure procedure, superior to L-LDN regarding donor safety. There has been an unacceptably high rate of reoperations in our L-LDN series but without mortality or significant sequelae. A careful look at some other L-LDN series also suggests increased morbidity/mortality. Our data do, however, support the view that a perfect, uncomplicated L-LDN appears to be the superior procedure, and the laparoscopic procedure is still evolving. Donor safety may be improved by avoiding obese donors, stapling of the renal artery (not clipping), and perhaps by hand assistance. Furthermore, we will consider the retroperitoneal approach. PMID- 15880078 TI - Recombinant human growth hormone treatment after liver transplantation in childhood: the 5-year outcome. AB - BACKGROUND: Because the results of short-term recombinant human growth hormone (rhGH) treatment in children with growth impairment after liver transplantation (LTx) have been promising, we have studied the long-term effects of rhGH on growth and graft function after LTx. METHODS: Indications for rhGH treatment were height standard deviation score (hSDS) below -2.0 or growth velocity SDS below 0 and LTx at least 18 months before inclusion. Eight growth-retarded children were treated with rhGH for more than 5 years. RESULTS: During the first year, median growth rate improved from 3.3 to 7.0 cm/year. In the second and third year, growth velocity remained high at 6.6 cm/year and 6.2 cm/year, respectively (P=0.008). In the fourth year, median growth velocity started to decline but still remained above baseline during the fifth year of treatment (4.2 cm/year). The median hSDS improved from -3.6 to -2.7. During the rhGH treatment, no acute rejection episodes were detected, and graft function remained stable in all except one patient. She was diagnosed with chronic rejection in the third year of rhGH treatment. The patient had elevated liver enzymes and abnormal liver function tests already before rhGH treatment. CONCLUSIONS: The efficacy of rhGH treatment is sustained after the first year in liver-transplant children with non GH-deficient growth retardation. Because of a potential risk of side effects, close monitoring of these patients is required. PMID- 15880079 TI - Living kidney donor follow-up in a dedicated clinic. AB - BACKGROUND: Long-term effects of uninephrectomy for kidney donation are of particular interest in the currently increasing practice of living-donor transplantation. We have retrospectively analyzed the general health status and renal and cardiovascular consequences of living-related kidney donation. METHODS: Data of living-related kidney donors who were regularly followed up in a dedicated clinic at the Sindh Institute of Urology and Transplantation between July 2000 and January 2004 was retrieved. They had donated their kidneys from 1986 onward. Data on weight, blood pressure, creatinine clearance, level of proteinuria, and new onset diabetes mellitus were analyzed. RESULTS: Seven hundred and thirty-six donors with a mean age of 36+/-10.9 years (M:F 1.1:1) were evaluated. With a mean postnephrectomy duration of 3+/-3.2 years (range 6 months 18 years), the creatinine clearance fell to 87% of prenephrectomy values, and 49 (6.7%) had a creatinine clearance of less than 60 mL/ min. Hypertension developed in 76 (10.3%) donors, and 179 (24.3%) had proteinuria exceeding 150 mg/24 hr. Overweight (27.8%) and obese subjects (11.5%) had a higher prevalence of hypertension and new onset diabetes mellitus. One donor developed end-stage renal failure. CONCLUSION: Donor nephrectomy has minimal adverse effects on overall health status. Regular donor follow-up identifies at-risk populations and potentially modifiable factors. PMID- 15880080 TI - Cognitive abilities in children after liver transplantation. AB - BACKGROUND: The authors investigated the cognitive status during the late postoperative phase in children who had undergone liver transplantation (LTx). METHODS: The authors examined 44 children who had undergone liver transplantation at their center. The children were 8.9+/-2.3 (mean+/-SD) years of age and had received the transplant 6.1+/-2.6 years previously. In 24 of the 44 children, a living-related transplantation had been carried out. Cognitive abilities were assessed with the three subscales of the Kaufman Assessment Battery for Children (K-ABC): the sequential processing scale (SES), the simultaneous processing scale (SIS), and the achievement scale (AS). RESULTS: The children scored below the population mean but within the normal range on all subscales of the K-ABC. In the SIS and the AS, age at transplantation influenced the cognitive outcome, as the children who were younger at transplantation scored significantly better than the older children and their results were within the normal range. However, for the SES, no such differences were found. A multiple regression analysis revealed that duration of illness and height at transplantation predicts the performance in the SIS and the AS. To a lesser degree, type of transplantation (cadaveric vs. living related) predicts performance in the AS. Performance in the SES was not predicted by any of these variables. Time since LTx and type of immunosuppressive regimen were not associated with the cognitive status after transplantation. CONCLUSIONS: Children who are younger, with a shorter duration of illness, and who are more physically developed before LTx have a better prognosis regarding their mental development. However, this result does not hold for sequential processing functions, which showed no relationship with any of these variables. This could indicate differential effects of liver disease and consecutive metabolic derangements on brain development during the preoperative phase. PMID- 15880081 TI - Impact of the expanded criteria donor allocation system on the use of expanded criteria donor kidneys. AB - BACKGROUND: The U.S. Organ Procurement and Transplantation Network recently implemented a policy allocating expanded criteria donor (ECD) kidneys by waiting time alone. ECD kidneys were defined as having a risk of graft failure > or = 1.7 times that of ideal donors. ECDs include any donor > or = 60 years old and donors 50 to 59 years old with at least two of the following: terminal creatinine >1.5 mg/dL, history of hypertension, or death by cerebrovascular accident. The impact of this policy on use of ECD kidneys is assessed. METHODS: The authors compared use of ECD kidneys recovered in the 18 months immediately before and after policy implementation. Differences were tested using t test and chi2 analyses. RESULTS: There was an 18.3% increase in ECD kidney recoveries and a 15.0% increase in ECD kidney transplants in the first 18 months after policy implementation. ECD kidneys made up 22.1% of all recovered kidneys and 16.8% of all transplants, compared with 18.8% (P<0.001) and 14.5% (P<0.001), respectively, in the prior period. The discard rate was unchanged. The median relative risk (RR) for graft failure for transplanted ECD kidneys was 2.07 versus 1.99 in the prepolicy period (P=not significant); the median RR for procured ECD kidneys was unchanged at 2.16. The percentage of transplanted ECD kidneys with cold ischemia times (CIT) <12 hr increased significantly; the corresponding percentage for CIT > or = 24 hr decreased significantly. CONCLUSIONS: The recent increase in ECD kidney recoveries and transplants appears to be related to implementation of the ECD allocation system. PMID- 15880082 TI - Low incidence of acute rejection after living-donor liver transplantation: immunologic analyses by mixed lymphocyte reaction using a carboxyfluorescein diacetate succinimidyl ester labeling technique. AB - BACKGROUND: To monitor antidonor alloreactivity for accurate diagnosis of acute rejection after living-donor liver transplantation (LDLT), we used a mixed lymphocyte reaction (MLR) assay using an intracellular fluorescent dye carboxyfluorescein diacetate succimidyl ester (CFSE)-labeling technique (CFSE MLR) in 29 consecutive patients who underwent adult-to-adult LDLT. METHODS: For patients who developed moderate or severe disorders in liver function, CFSE-MLR was performed together with needle biopsy of the liver allografts immediately after liver dysfunction had occurred. CFSE-labeled peripheral blood mononuclear cells (PBMC) from recipients and irradiated autologous, donor, or third-party PBMC were cultured, and then proliferation and CD25 expression in each of the CD4+ and CD8+ T cell subsets were analyzed by flow cytometry. RESULTS: Twelve (41.4%) of the 29 patients developed moderate or severe disorders in liver function within 6 months after LDLT. Eight of the 12 patients (overall incidence of 27.6%) suffering from liver function disorder were diagnosed on the basis of liver biopsy results as having mild or moderate acute rejection. However, only 4 of the 12 patients (overall incidence of 13.8%) showed remarkable proliferation of CD8+ T cells in association with CD25 expression on antidonor CFSE-MLR. The other eight patients were eventually diagnosed as having recurrence of original hepatitis, drug-induced hepatotoxicity, or congestion of the anterior segment of the liver allograft by further extensive examinations or in retrospect. CONCLUSIONS: The results of CFSE-MLR assays, which could be used for rigorously monitoring rejection, provided evidence of low incidence of acute rejection after LDLT. PMID- 15880083 TI - Single-antigen-expressing cell lines are excellent tools for detecting human leukocyte antigen-C-reactive antibodies in kidney transplant recipients. AB - BACKGROUND: Human leukocyte antigen (HLA)-C is expressed on nucleated cells and platelets in lower levels than HLA-A,B, and its antigens are in linkage disequilibrium with HLA-B antigens. Therefore, HLA-C antibody detection is difficult. The authors questioned whether HLA-C could serve as a target in clinical kidney transplantation using a newly developed assay. METHODS: Flow cytometry was performed with sera from patients (n=34) awaiting a kidney retransplant using nine cell lines expressing a single HLA-C antigen (single antigen lines [SAL]). RESULTS: The SAL were validated with HLA-C-specific alloantisera and human monoclonal antibodies against HLA-A, -B, and -C. The results were in agreement with the specificities previously reported. Exceptions, because of new HLA-C specificities used here, could be explained by epitope sharing between the antigens. With respect to patient sera, 15 of the 34 patients tested (44%) showed serum reactivity toward one or more HLA-C SAL. CONCLUSIONS: In contrast to peripheral blood lymphocytes, SAL are excellent targets for detecting HLA-C-reactive alloantibodies by flow cytometry. This preliminary analysis revealed that HLA-C-reactive antibodies are frequently present in sera of retransplant patients, serving as possible targets in clinical transplantation. PMID- 15880084 TI - PELD score and posttransplant outcome in pediatric liver transplantation: a retrospective study of 100 recipients. AB - BACKGROUND: Pediatric End-stage Liver Disease (PELD) score is proposed as an objective tool to prioritize children awaiting liver transplantation (LT), higher PELD being associated with increased pre-LT mortality. This study investigated whether PELD may also impact on post-LT results. METHODS: PELD was retrospectively analyzed in 100 pediatric recipients of a primary LT from living related (n = 49) or postmortem donors (PMD, n = 51). The main pre-LT diagnosis was biliary atresia (n = 64), hepatic malignancy and fulminant hepatitis cases being excluded. PELD was calculated in all patients at the time of pre-LT assessment. Considering the median delay of 117 days between listing and LT in the PMD subgroup, a second PELD was calculated at the time of LT, allowing the determination of a delta PELD during the waiting period. PMD grafts were allocated using an allocation system taking into account waiting times as well as medical urgency, operative at EuroTransplant. RESULTS: Overall 5-year actuarial patient and graft survivals were 96% and 91%, respectively. PELD at listing (13.3 +/- 9.7) showed a normal statistical distribution. PELD scores at listing and at LT were not found to significantly impact on post-LT outcome (NS). In contrast, higher delta PELD might be associated with lower posttransplant patient survival (p = 0.094). CONCLUSIONS: The results of this retrospective analysis suggest that giving priority to high PELD recipients may not result in worsening post-LT outcome. Accordingly, these data support such "sickest children first" allocation policy, which should contribute to reduce pre-LT mortality without worsening post LT results and increasing organ waste. PMID- 15880085 TI - Dehydroepiandrosterone (DHEA). AB - Health care providers are often asked about the efficacy and safety of this dietary supplement. PMID- 15880086 TI - ARBs and Myocardial Infarction. AB - A recent editorial in the British Journal suggested that ARBs used for treatment of hypertension may increase the risk of myocardial infarction. PMID- 15880087 TI - Albumin-bound paclitaxel (Abraxane) for advanced breast cancer. AB - A new formulation that should be safer and easier to administer than Taxol. PMID- 15880088 TI - Shigellosis. AB - Shigellosis is a global human health problem. Four species of Shigella i.e. S. dysenteriae, S. flexneri, S. boydii and S. sonnei are able to cause the disease. These species are subdivided into serotypes on the basis of O-specific polysaccharide of the LPS. Shigella dysenteriae type 1 produces severe disease and may be associated with life-threatening complications. The symptoms of shigellosis include diarrhoea and/or dysentery with frequent mucoid bloody stools, abdominal cramps and tenesmus. Shigella spp. cause dysentery by invading the colonic mucosa. Shigella bacteria multiply within colonic epithelial cells, cause cell death and spread laterally to infect and kill adjacent epithelial cells, causing mucosal ulceration, inflammation and bleeding. Transmission usually occurs via contaminated food and water or through person-to-person contact. Laboratory diagnosis is made by culturing the stool samples using selective/differential agar media. Shigella spp. are highly fragile organism and considerable care must be exercised in collecting faecal specimens, transporting them to the laboratories and in using appropriate media for isolation. Antimicrobial agents are the mainstay of therapy of all cases of shigellosis. Due to the global emergence of drug resistance, the choice of antimicrobial agents for treating shigellosis is limited. Although single dose of norfloxacin and ciprofloxacin has been shown to be effective, they are currently less effective against S. dysenteriae type 1 infection. Newer quinolones, cephalosporin derivatives, and azithromycin are the drug of choice. However, fluoroquinolone resistant S. dysenteriae type 1 infection have been reported. Currently, no vaccines against Shigella infection exist. Both live and subunit parenteral vaccine candidates are under development. Because immunity to Shigella is serotype-specific, the priority is to develop vaccine against S. dysenteriae type 1 and S. flexneri type 2a. Shigella species are important pathogens responsible for diarrhoeal diseases and dysentery occurring all over the world. The morbidity and mortality due to shigellosis are especially high among children in developing countries. A recent review of literature (Kotloff et al.,1999) concluded that, of the estimated 165 million cases of Shigella diarrhoea that occur annually, 99% occur in developing countries, and in developing countries 69% of episodes occur in children under five years of age. Moreover, of the ca.1.1 million deaths attributed to Shigella infections in developing countries, 60% of deaths occur in the under-five age group. Travellers from developed to developing regions and soldiers serving under field conditions are also at an increased risk to develop shigellosis. PMID- 15880089 TI - Archaeal diversity in tidal flat sediment as revealed by 16S rDNA analysis. AB - During the past ten years, Archaea have been recognized as a widespread and significant component of marine picoplankton assemblages. More recently, the presence of novel archaeal phylogenetic lineages has been discovered in coastal marine environments, freshwater lakes, polar seas, and deep-sea hydrothermal vents. Therefore, we conducted an investigation into the archaeal community existing in tidal flat sediment collected from Ganghwa Island, Korea. Phylogenetic analysis of archaeal 16S rDNA amplified directly from tidal flat sediment DNA revealed the presence of two major lineages, belonging to the Crenarchaeota (53.9%) and Euryarchaeota (46.1%) phyla. A total of 102 clones were then sequenced and analyzed by comprehensive phylogenetic analysis. The sequences determined in our samples were found to be closely related to the sequences of clones which had been previously obtained from a variety of marine environments. Archaeal clones exhibited higher similarities (83.25-100%) to sequences from other environments in the public database than did those (75.22-98.46%) of previously reported bacterial clones obtained from tidal flat sediment. The results of our study suggest that the archaeal community in tidal flat sediment is remarkably diverse. PMID- 15880090 TI - Sphingopyxis granuli sp. nov., a beta-glucosidase-producing bacterium in the family Sphingomonadaceae in alpha-4 subclass of the Proteobacteria. AB - Strain Kw07T, a Gram-negative, non-spore-forming, rod-shaped bacterium, was isolated from granules in an Up-flow Anaerobic Sludge Blanket (UASB) bioreactor used in the treatment of brewery wastewater. 16S rRNA gene sequence analysis revealed that strain Kw07T belongs to the alpha-4 subclass of the Proteobacteria, and the highest degree of sequence similarity was determined to be to Sphingopyxis macrogoltabida IFO 15033T(97.8%). Chemotaxonomic data revealed that strain Kw07T possesses a quinone system with the predominant compound Q-10, the predominant fatty acid C18:1 omega7c, and sphingolipids, all of which corroborated our assignment of the strain to the Sphingopyxis genus. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that strain Kw07T represents a distinct species. Based on these data, Kw07T (=KCTC 12209T=NBRC 100800T) should be classified as the type strain for a novel Sphingopyxis species, for which the name Sphingopyxis granuli sp. nov. has been proposed. PMID- 15880091 TI - The taxonomy of Psilocybe fagicola-complex. AB - Psilocybe fagicola comprises a complex of more than eight species, six of them in Mexico, and all of them possessing a long pseudorhiza, a characteristic not listed by Heim and Cailleux in 1959 in the original description of the type species, but described by Guzman in 1978 and 1983. The description of Psilocybe fagicola s.s. is here emended to include the length of the cheilocystidia of (6-) 12-20 (-30) microm, as well as the absence or scarcity of pleurocystidia. Psilocybe xalapensis and P. wassoniorum are considered to be synonymous with P. fagicola s.s. However, Psilocybe banderillensis and P. herrerae from Mexico, P. columbiana from Colombia, and P. keralensis from India are considered to be valid species within this complex. Moreover, P. novoxalapensis and P. teofilae, both from Mexico, are described as new species. Length of spores, presence or absence of pleurocystidia and their variations, and type of cheilocystidia constitute the principal defining characteristics of the species. Setaceous hyphae at the base of the stipe, as well as caulocystidia, lack taxonomic value, as do other morphological characteristics, including pileipellis and subpileipellis. A key to the eight considered species is also presented within the paper. PMID- 15880092 TI - Effects of sulfate concentration on the anaerobic dechlorination of polychlorinated biphenyls in estuarine sediments. AB - In order to determine the effects of sulfate concentration on the anaerobic dechlorination of polychlorinated biphenyls, sediments spiked with Aroclor 1242 were made into slurries using media which had various sulfate concentrations ranging from 3 to 23 mM. The time course of dechlorination clearly demonstrated that dechlorination was inhibited at high concentration of sulfate due to less dechlorination of meta-substituted congeners. When the dechlorination patterns were analyzed by the calculation of Euclidean distance, the dechlorination pathway in the 3 mM sulfate samples was found to be different from that observed in the 13 mM samples, although the extent of dechlorination in these two samples was similar. It is possible that the dechlorination in the high sulfate concentration samples is inhibited by the suppression of growth of methanogen, which have been shown to be meta-dechlorinating microorganisms. PMID- 15880093 TI - Molecular characterization of TEM-type beta-lactamases identified in cold-seep sediments of Edison Seamount (south of Lihir Island, Papua New Guinea). AB - To determine the prevalence and genotypes of beta-lactamases among clones of a metagenomic library from the cold-seep sediments of Edison seamount (10,000 years old), we performed pulse-field gel electrophoresis, antibiotic susceptibility testing, pI determination, and DNA sequencing analysis. Among the 8,823 clones of the library, thirty clones produced beta-lactamases and had high levels of genetic diversity. Consistent with minimum inhibitory concentration patterns, we found that five (16.7%) of thirty clones produced an extended-spectrum beta lactamase. 837- and 259-bp fragments specific to blaTEM genes were amplified, as determined by banding patterns of PCR amplification with designed primers. TEM-1 was the most prevalent beta-lactamase and conferred resistance to ampicillin, piperacillin, and cephalothin. TEM-116 had a spectrum that was extended to ceftazidime, cefotaxime, and aztreonam. The resistance levels conferred by the pre-antibiotic era alleles of TEM-type beta-lactamases were essentially the same as the resistance levels conferred by the TEM-type alleles which had been isolated from clinically resistant strains of bacteria of the antibiotic era. Our first report on TEM-type beta-lactamases of the pre-antibiotic era indicates that TEM-type beta-lactamases paint a picture in which most of the diversity of the enzymes may not be the result of recent evolution, but that of ancient evolution. PMID- 15880094 TI - Enhancement of gene delivery to cancer cells by a retargeted adenovirus. AB - The inefficiency of in vivo gene transfer using currently available vectors reflects a major hurdle in cancer gene therapy. Both viral and non-viral approaches that improve gene transfer efficiency have been described, but suffer from a number of limitations. Herein, a fiber-modified adenovirus, carrying the small peptide ligand on the capsid, was tested for the delivery of a transgene to cancer cells. The fiber-modified adenovirus was able to mediate the entry and expression of a beta-galactosidase into cancer cells with increased efficiency compared to the unmodified adenovirus. Particularly, the gene transfer efficiency was improved up to 5 times in OVCAR3 cells, an ovarian cancer cell line. Such transduction systems hold promise for delivering genes to transferrin receptor overexpressing cancer cells, and could be used for future cancer gene therapy. PMID- 15880095 TI - Staphylococcus aureus siderophore-mediated iron-acquisition system plays a dominant and essential role in the utilization of transferrin-bound iron. AB - Staphylococcus aureus is known to be capable of utilizing transferrin-bound iron, via both siderophore- and transferrin-binding protein (named IsdA)-mediated iron acquisition systems. This study was designed in order to determine which iron acquisition system plays the essential or dominant role with respect to the acquisition of iron from human transferrin, in the growth of S. aureus. Holotransferrin (HT) and partially iron-saturated transferrin (PT), but not apotransferrin (AT), were found to stimulate the growth of S. aureus. S. aureus consumed most of the transferrin-bound iron during the exponential growth phase. Extracellular proteases were not, however, involved in the liberation of iron from transferrin. Transferrin-binding to the washed whole cells via IsdA was not observed during the culture. The expression of IsdA was observed only in the deferrated media with AT, but not in the media supplemented with PT or HT. In contrast, siderophores were definitely produced in the deferrated media with PT and HT, as well as in the media supplemented with AT. The siderophores proved to have the ability to remove iron directly from transferrin, but the washed whole cells expressing IsdA did not. In the bioassay, the growth of S. aureus on transferrin-bound iron was stimulated by the siderophores alone. These results demonstrate that the siderophore-mediated iron-acquisition system plays a dominant and essential role in the uptake of iron from transferrin, whereas the IsdA-mediated iron-acquisition system may play only an ancillary role in the uptake of iron from transferrin. PMID- 15880096 TI - Functional characterization of the gene encoding UDP-glucose: tetrahydrobiopterin alpha-glucosyltransferase in Synechococcus sp. PCC 7942. AB - In this study, we attempted to characterize the Synechococcus sp. PCC 7942 mutant resultant from a disruption in the gene encoding UDP-glucose: tetrahydrobiopterin alpha-glucosyltransferase (BGluT). 2D-PAGE followed by MALDI-TOF mass spectrometry revealed that phycocyanin rod linker protein 33K was one of the proteins expressed at lower level in the BGluT mutant. BGluT mutant cells were also determined to be more sensitive to high light stress. This is because photosynthetic O2 exchange rates were significantly decreased, due to the reduced number of functional PSIs relative to the wild type cells. These results suggested that, in Synechococcus sp. PCC 7942, BH4-glucoside might be involved in photosynthetic photoprotection. PMID- 15880097 TI - Stabilities of artificially transconjugated plasmids for the bioremediation of cocontaminated sites. AB - Here, we attempted to evaluate the activity of artificially transconjugated multiple plasmids in "designer biocatalysts" for the bioremediation of cocontaminated sites under nonselective conditions. We observed profound losses in the percent survivals of artificially transconjugated plasmid activity (66-78% loss immediately after freeze-drying, 99.45-99.88% loss by the end of 6 months storage) in reconstituted Pseudomonas sp. KM12TC. Such unpredictable high losses of this particular plasmid appeared to clearly be a deleterious effect. However, even after 6 months of storage, the cells remained able to degrade 95% of phenol within 9 days, and the full efflux of 73As, as compared to that of the non-freeze dried cells, was successfully achieved 4 to 9 days later. These results indicate that "stable designer biocatalysts" can remain viable, even after freeze-drying and 6 months of storage. PMID- 15880098 TI - Identification of non-mutans streptococci organisms in dental plaques recovering on mitis-salivarius bacitracin agar medium. AB - The objective of this study was to both isolate and identify non-mutans streptococci organisms (non-MSO) from dental plaques recovered on mitis salivarius sucrose bacitracin agar (MSB) plates. The dental plaque samples, which had been collected from 63 human subjects, were diluted and plated on MSB. The bacteria growing on the MSB plates were then identified with biochemical tests, as well as with 16S rDNA cloning and sequencing techniques. Our data indicated that bacteria from 30 subjects had been recovered on the MSB plates. Among the 21 typical colonies selected from the 30 subjects, 12 colonies, derived from 10 subjects, were identified as non-MSO. These 12 colonies were determined to be Streptococcus anginosus (8 colonies), S. sanguinis (1 colony), and Pantoea agglomerans (3 colonies). These results strongly suggest that a new selective medium will be required for the reliable isolation of mutans streptococci. PMID- 15880099 TI - Identification of Actinobacillus actinomycetemcomitans using species-specific 16S rDNA primers. AB - The purpose of this study was to develop species-specific PCR primers for use in the identification and detection of Actinobacillus actinomycetemcomitans. These primers target variable regions of the 16S ribosomal RNA coding gene (rDNA). We assessed the specificity of the primers against 9 A. actinomycetemcomitans strains and 11 strains (3 species) of the Haemophilus genus. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC 33384T. Our obtained data revealed that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 4 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these PCR primers are incredibly sensitive, and should prove suitable for application in epidemiological studies, as well as the diagnosis and monitoring of periodontal pathogens after treatment for periodontitis. PMID- 15880100 TI - Molecular taxonomy of a soil actinomycete isolate, KCCM10454 showing neuroprotective activity by 16S rRNA and rpoB gene analysis. AB - Epilepsy constitutes a significant public health problem, and even the newest drugs and neurosurgical techniques have proven unable to cure the disease. In order to select a group of isolates which could generate an active compound with neuroprotective or antiepileptic properties, we isolated 517 actinomycete strains from soil samples taken from Jeju Island, in South Korea. We then screened these strains for possible anti-apoptotic effects against serum deprivation-induced hippocampal cell death, using the 3-(4, 5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay as an in vitro test. The excitotoxic glutamate analog, kainic acid (KA), was used to induce seizures in experimental mice in our in vivo tests. As a result of this testing, we located one strain which exhibited profound neuroprotective activity. This strain was identified as a Streptomyces species, and exhibited the rifampin-resistant genotype, Asn(AAC)442, according to the results of 16S rRNA and rpoB gene analyses. PMID- 15880101 TI - Interchromosomal associations between alternatively expressed loci. AB - The T-helper-cell 1 and 2 (T(H)1 and T(H)2) pathways, defined by cytokines interferon-gamma (IFN-gamma) and interleukin-4 (IL-4), respectively, comprise two alternative CD4+ T-cell fates, with functional consequences for the host immune system. These cytokine genes are encoded on different chromosomes. The recently described T(H)2 locus control region (LCR) coordinately regulates the T(H)2 cytokine genes by participating in a complex between the LCR and promoters of the cytokine genes Il4, Il5 and Il13. Although they are spread over 120 kilobases, these elements are closely juxtaposed in the nucleus in a poised chromatin conformation. In addition to these intrachromosomal interactions, we now describe interchromosomal interactions between the promoter region of the IFN-gamma gene on chromosome 10 and the regulatory regions of the T(H)2 cytokine locus on chromosome 11. DNase I hypersensitive sites that comprise the T(H)2 LCR developmentally regulate these interchromosomal interactions. Furthermore, there seems to be a cell-type-specific dynamic interaction between interacting chromatin partners whereby interchromosomal interactions are apparently lost in favour of intrachromosomal ones upon gene activation. Thus, we provide an example of eukaryotic genes located on separate chromosomes associating physically in the nucleus via interactions that may have a function in coordinating gene expression. PMID- 15880102 TI - CtBP3/BARS drives membrane fission in dynamin-independent transport pathways. AB - Membrane fission is a fundamental step in membrane transport. So far, the only fission protein machinery that has been implicated in in vivo transport involves dynamin, and functions in several, but not all, transport pathways. Thus, other fission machineries may exist. Here, we report that carboxy-terminal binding protein 3/brefeldin A-ribosylated substrate (CtBP3/BARS) controls fission in basolateral transport from the Golgi to the plasma membrane and in fluid-phase endocytosis, whereas dynamin is not involved in these steps. Conversely, CtBP3/BARS protein is inactive in apical transport to the plasma membrane and in receptor-mediated endocytosis, both steps being controlled by dynamin. This indicates that CtBP3/BARS controls membrane fission in endocytic and exocytic transport pathways, distinct from those that require dynamin. PMID- 15880103 TI - Human recombination hot spots hidden in regions of strong marker association. AB - The fine-scale distribution of meiotic recombination events in the human genome can be inferred from patterns of haplotype diversity in human populations but directly studied only by high-resolution sperm typing. Both approaches indicate that crossovers are heavily clustered into narrow recombination hot spots. But our direct understanding of hot-spot properties and distributions is largely limited to sperm typing in the major histocompatibility complex (MHC). We now describe the analysis of an unremarkable 206-kb region on human chromosome 1, which identified localized regions of linkage disequilibrium breakdown that mark the locations of sperm crossover hot spots. The distribution, intensity and morphology of these hot spots are markedly similar to those in the MHC. But we also accidentally detected additional hot spots in regions of strong association. Coalescent analysis of genotype data detected most of the hot spots but showed significant differences between sperm crossover frequencies and historical recombination rates. This raises the possibility that some hot spots, particularly those in regions of strong association, may have evolved very recently and not left their full imprint on haplotype diversity. These results suggest that hot spots could be very abundant and possibly fluid features of the human genome. PMID- 15880104 TI - Large-scale in vivo flux analysis shows rigidity and suboptimal performance of Bacillus subtilis metabolism. AB - Qualitative theoretical approaches such as graph theory and stoichiometric analyses are beginning to uncover the architecture and systemic functions of complex metabolic reaction networks. At present, however, only a few, largely unproven quantitative concepts propose functional design principles of the global flux distribution. As operational units of function, molecular fluxes determine the systemic cell phenotype by linking genes, proteins and metabolites to higher level biological functions. In sharp contrast to other 'omics' analyses, 'fluxome' analysis remained tedious. By large-scale quantification of in vivo flux responses, we identified a robust flux distribution in 137 null mutants of Bacillus subtilis. On its preferred substrate, B. subtilis has suboptimal metabolism because regulators of developmental programs maintain a 'standby' mode that invests substantial resources in anticipation of changing environmental conditions at the expense of optimal growth. Network rigidity and robustness are probably universal functional design principles, whereas the standby mode may be more specific. PMID- 15880105 TI - Laminin stimulates and guides axonal outgrowth via growth cone myosin II activity. AB - Guidance cues and signal transduction mechanisms acting at the nerve growth cone are fairly well understood, but the intracellular mechanisms operating to change the direction of axon outgrowth remain unknown. We now show that growth cones integrate myosin II-dependent contraction for rapid, coordinated turning at borders of laminin stripes in response to signals from laminin-activated integrin receptors; in the absence of myosin II activity, outgrowth continues across the borders. PMID- 15880106 TI - An fMRI investigation of race-related amygdala activity in African-American and Caucasian-American individuals. AB - Functional magnetic resonance imaging (fMRI) was used to examine the nature of amygdala sensitivity to race. Both African-American and Caucasian-American individuals showed greater amygdala activity to African-American targets than to Caucasian-American targets, suggesting that race-related amygdala activity may result from cultural learning rather than from the novelty of other races. Additionally, verbal encoding of African-American targets produced significantly less amygdala activity than perceptual encoding of African-American targets. PMID- 15880107 TI - Reversible blockade of experience-dependent plasticity by calcineurin in mouse visual cortex. AB - Numerous protein kinases have been implicated in visual cortex plasticity, but the role of serine/threonine protein phosphatases has not yet been established. Calcineurin, the only known Ca2+/calmodulin-activated protein phosphatase in the brain, has been identified as a molecular constraint on synaptic plasticity in the hippocampus and on memory. Using transgenic mice overexpressing calcineurin inducibly in forebrain neurons, we now provide evidence that calcineurin is also involved in ocular dominance plasticity. A transient increase in calcineurin activity is found to prevent the shift of responsiveness in the visual cortex following monocular deprivation, and this effect is reversible. These results imply that the balance between protein kinases and phosphatases is critical for visual cortex plasticity. PMID- 15880108 TI - 5-HTTLPR polymorphism impacts human cingulate-amygdala interactions: a genetic susceptibility mechanism for depression. AB - Carriers of the short allele of a functional 5' promoter polymorphism of the serotonin transporter gene have increased anxiety-related temperamental traits, increased amygdala reactivity and elevated risk of depression. Here, we used multimodal neuroimaging in a large sample of healthy human subjects to elucidate neural mechanisms underlying this complex genetic association. Morphometrical analyses showed reduced gray matter volume in short-allele carriers in limbic regions critical for processing of negative emotion, particularly perigenual cingulate and amygdala. Functional analysis of those regions during perceptual processing of fearful stimuli demonstrated tight coupling as a feedback circuit implicated in the extinction of negative affect. Short-allele carriers showed relative uncoupling of this circuit. Furthermore, the magnitude of coupling inversely predicted almost 30% of variation in temperamental anxiety. These genotype-related alterations in anatomy and function of an amygdala-cingulate feedback circuit critical for emotion regulation implicate a developmental, systems-level mechanism underlying normal emotional reactivity and genetic susceptibility for depression. PMID- 15880109 TI - SRF mediates activity-induced gene expression and synaptic plasticity but not neuronal viability. AB - Synaptic activity-dependent gene expression is critical for certain forms of neuronal plasticity and survival in the mammalian nervous system, yet the mechanisms by which coordinated regulation of activity-induced genes supports neuronal function is unclear. Here, we show that deletion of serum response factor (SRF) in specific neuronal populations in adult mice results in profound deficits in activity-dependent immediate early gene expression, but components of upstream signaling pathways and cyclic AMP-response element binding protein (CREB)-dependent transactivation remain intact. Moreover, SRF-deficient CA1 pyramidal neurons show attenuation of long-term synaptic potentiation, a model for neuronal information storage. Furthermore, in contrast to the massive neurodegeneration seen in adult mice lacking CREB family members, SRF-deficient adult neurons show normal morphologies and basal excitatory synaptic transmission. These findings indicate that the transcriptional events underlying neuronal survival and plasticity are dissociable and that SRF plays a prominent role in use-dependent modification of synaptic strength in the adult brain. PMID- 15880110 TI - XTRPC1-dependent chemotropic guidance of neuronal growth cones. AB - Calcium arising through release from intracellular stores and from influx across the plasma membrane is essential for signalling by specific guidance cues and by factors that inhibit axon regeneration. The mediators of calcium influx in these cases are largely unknown. Transient receptor potential channels (TRPCs) belong to a superfamily of Ca2+-permeable, receptor-operated channels that have important roles in sensing and responding to changes in the local environment. Here we report that XTRPC1, a Xenopus homolog of mammalian TRPC1, is required for proper growth cone turning responses of Xenopus spinal neurons to microscopic gradients of netrin-1, brain-derived neurotrophic factor and myelin-associated glycoprotein, but not to semaphorin 3A. Furthermore, XTRPC1 is required for midline guidance of axons of commissural interneurons in the developing Xenopus spinal cord. Thus, members of the TRPC family may serve as a key mediator for the Ca2+ influx that regulates axon guidance during development and inhibits axon regeneration in adulthood. PMID- 15880111 TI - Geometric and functional organization of cortical circuits. AB - Can neuronal morphology predict functional synaptic circuits? In the rat barrel cortex, 'barrels' and 'septa' delineate an orderly matrix of cortical columns. Using quantitative laser scanning photostimulation we measured the strength of excitatory projections from layer 4 (L4) and L5A to L2/3 pyramidal cells in barrel- and septum-related columns. From morphological reconstructions of excitatory neurons we computed the geometric circuit predicted by axodendritic overlap. Within most individual projections, functional inputs were predicted by geometry and a single scale factor, the synaptic strength per potential synapse. This factor, however, varied between projections and, in one case, even within a projection, up to 20-fold. Relationships between geometric overlap and synaptic strength thus depend on the laminar and columnar locations of both the pre- and postsynaptic neurons, even for neurons of the same type. A large plasticity potential appears to be incorporated into these circuits, allowing for functional 'tuning' with fixed axonal and dendritic arbor geometry. PMID- 15880112 TI - Recruitment of adult thymic progenitors is regulated by P-selectin and its ligand PSGL-1. AB - The molecular mechanisms that direct the migration of early T lymphocyte progenitors to the thymus are unknown. We show here that P-selectin is expressed by thymic endothelium and that lymphoid progenitors in bone marrow and thymus bind P-selectin. Parabiosis, competitive thymus reconstitution and short-term homing assays indicated that P-selectin and its ligand PSGL-1 are functionally important components of the thymic homing process. Accordingly, thymi of mice lacking PSGL-1 contained fewer early thymic progenitors and had increased empty niches for prothymocytes compared with wild-type mice. Furthermore, the number of resident thymic progenitors controls thymic expression of P-selectin, suggesting that regulation of P-selectin expression by a thymic 'niche occupancy sensor' may be used to direct progenitor access. PMID- 15880113 TI - Cyclic beta-1,2-glucan is a Brucella virulence factor required for intracellular survival. AB - Pathogenic brucella bacteria have developed strategies to persist for prolonged periods of time in host cells, avoiding innate immune responses. Here we show that the cyclic beta-1,2-glucans (CbetaG) synthesized by brucella is important for circumventing host cell defenses. CbetaG acted in lipid rafts found on host cell membranes. CbetaG-deficient mutants failed to prevent phagosome-lysosome fusion and could not replicate. However, when treated with purified CbetaG or synthetic methyl-beta-cyclodextrin, the mutants were able to control vacuole maturation by avoiding lysosome fusion, and this allowed intracellular brucella to survive and reach the endoplasmic reticulum. Fusion between the endoplasmic reticulum and the brucella-containing vacuole depended on the brucella virulence type IV secretion system but not on CbetaG. Brucella CbetaG is thus a virulence factor that interacts with lipid rafts and contributes to pathogen survival. PMID- 15880114 TI - Breakdown of avalanche critical behaviour in polycrystalline plasticity. AB - Acoustic emission experiments on creeping ice as well as numerical simulations argue for a self-organization of collective dislocation dynamics during plastic deformation of single crystals into a scale-free pattern of dislocation avalanches characterized by intermittency, power-law distributions of avalanche sizes, complex space-time correlations and aftershock triggering. Here, we address the question of whether such scale-free, close-to-critical dislocation dynamics will still apply to polycrystals. We show that polycrystalline plasticity is also characterized by intermittency and dislocation avalanches. However, grain boundaries hinder the propagation of avalanches, as revealed by a finite (grain)-size effect on avalanche size distributions. We propose that the restraint of large avalanches builds up internal stresses that push temporally the dynamical system into a supercritical state, off the scale-invariant critical regime, and trigger secondary avalanches in neighbouring grains. This modifies the statistical properties of the avalanche population. The results might also bring into question the classical ways of modelling plasticity in polycrystalline materials, based on homogenization procedures. PMID- 15880115 TI - Photomechanical actuation in polymer-nanotube composites. AB - For some systems, energy from an external source can trigger changes in the internal state of the structure, leading to a mechanical response much larger than the initial input. The ability to unlock this internal work in a solid-state structure is of key importance for many potential applications. We report a novel phenomenon of photo-induced mechanical actuation observed in a polymer-nanotube composite when exposed to infrared radiation. At small strains the sample tends to expand, when stimulated by photons, by an amount that is orders of magnitude greater than the pristine polymer. Conversely, at larger applied pre-strain, it will contract under identical infrared excitation. The behaviour is modelled as a function of orientational ordering of nanotubes induced by the uniaxial extension. It is thought that no other materials can display this continuously reversible response of so large a magnitude, making rubber nanocomposites important for actuator applications. PMID- 15880116 TI - A practical route for the fabrication of large single-crystal (RE)-Ba-Cu-O superconductors. AB - Single-crystal superconductors of the general formula (LRE)-Ba-Cu-O (light rare earth, LRE = Nd, Sm, Eu and Gd) have considerable potential for engineering applications because of their ability to trap magnetic fields significantly higher than those achievable with permanent magnets. But the lack of a process by which these materials can be fabricated reliably and economically in the form of large single grains has severely hindered their development. We report a practical processing method for the fabrication in air of single-crystal (RE)BCO. The technique is economical and offers considerable freedom in terms of the processing parameters and reproducibility in growth of oriented single grains. The process is based primarily on the development of a new type of generic seed crystal that can effectively promote the epitaxial nucleation of any (RE)BCO system, and secondly on suppressing the formation of RE-Ba solid solution in a controlled manner within large grains processed in air. PMID- 15880117 TI - Quantum dots spectrally distinguish multiple species within the tumor milieu in vivo. AB - A solid tumor is an organ composed of cancer and host cells embedded in an extracellular matrix and nourished by blood vessels. A prerequisite to understanding tumor pathophysiology is the ability to distinguish and monitor each component in dynamic studies. Standard fluorophores hamper simultaneous intravital imaging of these components. Here, we used multiphoton microscopy techniques and transgenic mice that expressed green fluorescent protein, and combined them with the use of quantum dot preparations. We show that these fluorescent semiconductor nanocrystals can be customized to concurrently image and differentiate tumor vessels from both the perivascular cells and the matrix. Moreover, we used them to measure the ability of particles of different sizes to access the tumor. Finally, we successfully monitored the recruitment of quantum dot-labeled bone marrow-derived precursor cells to the tumor vasculature. These examples show the versatility of quantum dots for studying tumor pathophysiology and creating avenues for treatment. PMID- 15880118 TI - TLR activation triggers the rapid differentiation of monocytes into macrophages and dendritic cells. AB - Leprosy enables investigation of mechanisms by which the innate immune system contributes to host defense against infection, because in one form, the disease progresses, and in the other, the infection is limited. We report that Toll-like receptor (TLR) activation of human monocytes induces rapid differentiation into two distinct subsets: DC-SIGN+ CD16+ macrophages and CD1b+ DC-SIGN- dendritic cells. DC-SIGN+ phagocytic macrophages were expanded by TLR-mediated upregulation of interleukin (IL)-15 and IL-15 receptor. CD1b+ dendritic cells were expanded by TLR-mediated upregulation of granulocyte-macrophage colony-stimulating factor (GM CSF) and its receptor, promoted T cell activation and secreted proinflammatory cytokines. Whereas DC-SIGN+ macrophages were detected in lesions and after TLR activation in all leprosy patients, CD1b+ dendritic cells were not detected in lesions or after TLR activation of peripheral monocytes in individuals with the progressive lepromatous form, except during reversal reactions in which bacilli were cleared by T helper type 1 (TH1) responses. In tuberculoid lepromatous lesions, DC-SIGN+ cells were positive for macrophage markers, but negative for dendritic cell markers. Thus, TLR-induced differentiation of monocytes into either macrophages or dendritic cells seems to crucially influence effective host defenses in human infectious disease. PMID- 15880119 TI - Reversing systemic inflammatory response syndrome with chemokine receptor pepducins. AB - We describe a new therapeutic approach for the treatment of lethal sepsis using cell-penetrating lipopeptides-termed pepducins-that target either individual or multiple chemokine receptors. Interleukin-8 (IL-8), a ligand for the CXCR1 and CXCR2 receptors, is the most potent endogenous proinflammatory chemokine in sepsis. IL-8 levels rise in blood and lung fluids to activate neutrophils and other cells, and correlate with shock, lung injury and high mortality. We show that pepducins derived from either the i1 or i3 intracellular loops of CXCR1 and CXCR2 prevent the IL-8 response of both receptors and reverse the lethal sequelae of sepsis, including disseminated intravascular coagulation and multi-organ failure in mice. Conversely, pepducins selective for CXCR4 cause a massive leukocytosis that does not affect survival. CXCR1 and CXCR2 pepducins conferred nearly 100% survival even when treatment was postponed, suggesting that our approach might be beneficial in the setting of advanced disease. PMID- 15880120 TI - Delay of HIV-1 rebound after cessation of antiretroviral therapy through passive transfer of human neutralizing antibodies. AB - To determine the protective potential of the humoral immune response against HIV 1 in vivo we evaluated the potency of three neutralizing antibodies (2G12, 2F5 and 4E10) in suppressing viral rebound in six acutely and eight chronically HIV-1 infected individuals undergoing interruption of antiretroviral treatment (ART). Only two of eight chronically infected individuals showed evidence of a delay in viral rebound during the passive immunization. Rebound in antibody-treated acutely infected individuals upon cessation of ART was substantially later than in a control group of 12 individuals with acute infection. Escape mutant analysis showed that the activity of 2G12 was crucial for the in vivo effect of the neutralizing antibody cocktail. By providing further direct evidence of the potency, breadth and titers of neutralizing antibodies that are required for in vivo activity, these data underline both the potential and the limits of humoral immunity in controlling HIV-1 infection. PMID- 15880121 TI - Domain structure of separase and its binding to securin as determined by EM. AB - After the degradation of its inhibitor securin, separase initiates chromosome segregation during the metaphase-to-anaphase transition by cleaving cohesin. Here we present a density map at a resolution of 25 A of negatively stained separase securin complex. Based on labeling data and sequence analysis, we propose a model for the structure of separase, consisting of 26 ARM repeats, an unstructured region of 280 residues and two caspase-like domains, with securin binding to the ARM repeats. PMID- 15880122 TI - Versatile modes of peptide recognition by the AAA+ adaptor protein SspB. AB - Energy-dependent proteases often rely on adaptor proteins to modulate substrate recognition. The SspB adaptor binds peptide sequences in the stress-response regulator RseA and in ssrA-tagged proteins and delivers these molecules to the AAA+ ClpXP protease for degradation. The structure of SspB bound to an ssrA peptide is known. Here, we report the crystal structure of a complex between SspB and its recognition peptide in RseA. Notably, the RseA sequence is positioned in the peptide-binding groove of SspB in a direction opposite to the ssrA peptide, the two peptides share only one common interaction with the adaptor, and the RseA interaction site is substantially larger than the overlapping ssrA site. This marked diversity in SspB recognition of different target proteins indicates that it is capable of highly flexible and dynamic substrate delivery. PMID- 15880123 TI - ATP hydrolysis cycle-dependent tail motions in cytoplasmic dynein. AB - The motor protein dynein is predicted to move the tail domain, a slender rod-like structure, relative to the catalytic head domain to carry out its power stroke. Here, we investigated ATP hydrolysis cycle-dependent conformational dynamics of dynein using fluorescence resonance energy transfer analysis of the dynein motor domain labeled with two fluorescent proteins. We show that dynein adopts at least two conformational states (states I and II), and the tail undergoes ATP-induced motions relative to the head domain during transitions between the two states. Our measurements also suggest that in the course of the ATP hydrolysis cycle of dynein, the tail motion from state I to state II takes place in the ATP-bound state, whereas the motion from state II to state I occurs in the ADP-bound state. The latter tail motion may correspond to the predicted power stroke of dynein. PMID- 15880125 TI - The relationship between mouthrinse matrix metalloproteinases (MMP-1, 8, 13) and albumin levels with the degree of oral mucositis in allogeneic stem cell transplant patients. AB - Our aim was to examine the relationship between mouthrinse matrix metalloproteinases (MMPs) and whole albumin levels (AL) relative to oral mucositis (OM) in allogeneic stem cell transplant (alloSCT) patients. Mouthrinse vertebrate collagenase levels are positively correlated with connective tissue destruction (CTD) in periodontitis and may also be involved in CTD associated with OM. Increases in salivary AL have been noted prior to OM onset and may serve as a predictive tool for OM and as a positive control in this study. A total of 23 alloSCT patients were visited eight times over 4 weeks following the transplant. OM was scored via a previously validated examiner-based ordinal system. Mouthrinse samples were collected and analyzed for MMP-1, 8, 13 (members of the vertebrate collagenase group) and AL. No significant correlation was found for MMP levels relative to OM scores. AL were positively and significantly associated with OM scores (P<0.001). MMP levels may not be an important factor in OM development and severity; however, mouthrinse AL may serve as a more objective measure of OM development and severity. PMID- 15880126 TI - The mechanism for low yield of blood culture in invasive aspergillosis; the clinical importance of antigen detection tests revisited. PMID- 15880127 TI - Longitudinal follow-up of body composition in hematopoietic stem cell transplant patients. AB - Patients with hematological malignancies are well nourished prior to allogeneic hematopoietic stem cell transplantation (HSCT). HSCT and associated complications can affect body composition. The study evaluated cross-sectionally the prevalence and longitudinally the changes in lean body mass index (LBMI) in HSCT patients. Patients (n=82) were classified as normal or low LBMI. Logistic regression analyses were used to estimate odds ratios (OR) for low vs normal LBMI, between healthy volunteers and patients; for limited or extensive vs no chronic graft versus-host-disease (GVHD); and for decreased (Karnofsky <80) vs normal functional status (>80). Patients were significantly more likely to have low LBMI at 6, 12 months, 2-3, 4-6 and >6 years than volunteers. In all, 38% of patients were below pre-HSCT LBMI at 4-6 years post-HSCT. Low LBMI was significantly associated with steroid treatment (OR 2.6, confidence intervals (CI) 1.3-5.2, P=0.008); limited (OR 5.5, CI 1.7-18.5, P=0.005) or extensive chronic GVHD (OR 20.3, CI 5.7-71.6, P<0.001); and decreased performance status (Karnofsky scores of < or =80) (OR 2.7, CI 1.3-5.9, P=0.01). Patients were more likely to have low LBMI than volunteers. Chronic GVHD and low performance status were associated with low LBMI; thus, complications and/or treatment increase the likelihood of low LBMI. PMID- 15880128 TI - BAVC regimen and autologous bone marrow transplantation for APL patients in second molecular remission: updated results. PMID- 15880129 TI - Pegfilgrastim after high-dose chemotherapy and autologous peripheral blood stem cell transplant: phase II study. AB - Pegfilgrastim is equivalent to daily filgrastim after standard dose chemotherapy in decreasing the duration of neutropenia. Daily filgrastim started within 1-4 days after autologous stem cell transplant (ASCT) leads to significant decrease in time to neutrophil engraftment. We undertook a study of pegfilgrastim after high-dose chemotherapy (HDC) and ASCT. In all, 38 patients with multiple myeloma or lymphoma, eligible to undergo HDC and ASCT, were enrolled. Patients received a single dose of 6 mg pegfilgrastim subcutaneously 24 h after ASCT. There were no adverse events secondary to pegfilgrastim. All patients engrafted neutrophils and platelets with a median of 10 and 18 days, respectively. The incidence of febrile neutropenia was 49% (18/37). Neutrophil engraftment results were compared to a historical cohort of patients who received no growth factors or prophylactic filgrastim after ASCT. Time to neutrophil engraftment using pegfilgrastim was comparable to daily filgrastim and was shorter than in a historical group receiving no filgrastim (10 vs 13.7 days, P<0.001). Pegfilgrastim given as a single fixed dose of 6 mg appears to be safe after HDC and ASCT. It accelerates neutrophil engraftment comparable to daily filgrastim after ASCT. Pegfilgrastim may be convenient to use in outpatient transplant units. PMID- 15880130 TI - Allogeneic peripheral stem-cell transplantation with reduced-intensity conditioning regimen in refractory primary B-cell prolymphocytic leukemia: a long term follow-up. PMID- 15880131 TI - Resolution of atopic dermatitis following allogeneic bone marrow transplantation for chronic myelogenous leukemia. PMID- 15880132 TI - Hyponatremia as the presenting feature of cerebral toxoplasmosis. PMID- 15880133 TI - Cost of autologous peripheral blood stem cell transplantation: the Norwegian experience from a multicenter cost study. AB - High-dose therapy with autologous blood progenitor cell support is now routinely used for patients with certain malignant lymphomas and multiple myeloma. We performed a prospective cost analysis of the mobilization, harvesting and cryopreservation phases and the high-dose therapy with stem cell reinfusion and hospitalization phases. In total, 40 consecutive patients were studied at four different university hospitals between 1999 and 2001. Data on direct costs were obtained on a daily basis. Data on indirect costs were allocated to the specific patient based on estimates of relevant department costs (ie the service department's costs), and by means of predefined allocation keys. All cost data were calculated at 2001 prices. The mean total costs for the two phases were US$ 32,160 (range US$ 19,092-50,550). The mean total length of hospital stay for two phases was 31 days (range 27-37). A large part of the actual cost in the harvest phase was attributed to stem cell mobilization, including growth factors, harvesting and cryopreservation. In the high-dose chemotherapy phase, the most significant part of the costs was nursing staff. Average total costs were considerably higher than actual DRG-based reimbursement from the government, indicating that the treatment of these patients was heavily subsidized by the basic hospital grants. PMID- 15880134 TI - Pneumonia caused by Trichosporon pullulans in an autologous peripheral blood stem cell transplant recipient: possible misidentification. PMID- 15880135 TI - Activation of epithelial CD98 glycoprotein perpetuates colonic inflammation. AB - Anomalies in the regulation and function of integrins have been implicated in the etiology of various pathologic conditions, including inflammatory disorders such as irritable bowel disease. Several classes of cell surface glycoproteins such as CD98 have been shown to play roles in integrins-mediated events. Here, we investigated the role of CD98 in intestinal inflammation using both in vivo and in vitro approaches. We found that in Caco2-BBE monolayers and colonic tissues, expression of CD98 was upregulated by the proinflammatory cytokine, interferon gamma (INF gamma). Furthermore, CD98 was highly upregulated in colonic tissues from mice with active colitis induced by dextran sodium sulfate (DSS), but not in DSS-treated INF gamma -/- mice. Administration of an anti-CD98 antibody worsened DSS-induced colitis in mice but had no effect on untreated control mice. Finally, we used Caco2-BBE cell monolayers to model intestinal epithelial wound healing, and found that activation of epithelial CD98 in DSS-treated monolayers inhibited monolayer reconstitution, but had no affect on untreated control monolayers. Our data collectively indicate that (i) CD98 upregulation is mediated by INF gamma during intestinal inflammation and (ii) activation of epithelial CD98 protein aggravates intestinal inflammation by reducing intestinal epithelial reconstitution. Overall, our data suggest that epithelial CD98 plays an important role in the perpetuation of intestinal inflammation. PMID- 15880136 TI - Molecular characterization of salivary gland malignancy using the Smgb-Tag transgenic mouse model. AB - The molecular mechanisms underlying salivary gland tumorigenesis remain unclear. In order to identify genetic changes that occur during the development of invasive adenocarcinoma from normal salivary gland, we used the Smgb-Tag transgenic mouse model. This transgene induces the progressive development of dysplasia to invasive adenocarcinoma in the submandibular salivary gland. Gene expression patterns from 20 submandibular glands (two normal, nine dysplasia and nine adenocarcinoma samples) were assessed using a mouse 15 K cDNA array. Unsupervised hierarchical clustering was used to group gene expression based on 157 differentially expressed genes distinguishing between dysplasias and adenocarcinomas. Further analysis identified 25 significantly overexpressed and 28 underexpressed cDNA sequences in adenocarcinoma as compared to dysplasia. Differential expression of five genes (Lcn2, Ptn, Cd24a, Mapk6 and Rnps1) was validated by quantitative real-time RT-PCR in a total of 48 mouse salivary gland tissues (seven histologically normal, 13 dysplasias and 28 adenocarcinomas), including the 20 samples analyzed by cDNA arrays. Immunohistochemical analysis was used to validate the expression of Ptn and Cd24a at the protein level in a subset of 16 mouse salivary glands (four normal, five dysplasia and seven adenocarcinoma samples), as well as in 23 human submandibular gland tumors (16 pleomorphic adenomas, three adenoid cystic carcinomas, one acinic cell carcinoma, one adenocarcinoma NOS, one myoepithelial and one mucoepidermoid carcinoma). We thus demonstrated that the Smgb-Tag transgenic mouse model is a useful tool for the identification of genes that are deregulated in salivary gland adenocarcinomas. Our data suggest that Ptn and Cd24a may be genetic markers associated with salivary gland tumorigenesis and/or progression. PMID- 15880137 TI - Loss of blood group A antigen expression in bladder cancer caused by allelic loss and/or methylation of the ABO gene. AB - Loss of ABO blood group antigen expression has been reported in transitional cell carcinoma (TCC) of the bladder. Synthesis of the ABO blood group antigen was genetically determined by allelic variants of the ABO gene assigned on 9q34.1. We analyzed loss of heterozygosity (LOH) and promoter hypermethylation of the ABO gene in TCC and compared them with alterations of A antigen expression in TCC, dysplasia and normal urothelium. A total of 81 samples of TCC of the bladder obtained from transurethral resection (TUR) (n=44) and radical cystectomy (n=37) were examined. Expression of the A antigen was evaluated by immunohistochemical staining (IHC) using anti-A antigen monoclonal antibody. LOH of the ABO gene locus was examined by blunt-end single-strand DNA conformational polymorphism (SSCP) analysis using flouresence-based auto sequencer. Promoter hypermethylation of the ABO gene were examined by bisulfite PCR-SSCP (BiPS) analysis and/or methylation-specific PCR (MSP). Loss of A allele and/or hypermethylation were significantly associated with abnormal expression of the A antigen in cases undergoing TUR (P=0.02) and radical cystectomy (P=0.0005). For the analysis of the concomitant dysplasia in 23 cases with TCC of the bladder, the expression of the A antigen was maintained, regardless of the A allelic loss or methylation status in the tumor. In conclusion, A allelic loss and hypermethylation in the promoter region of the ABO gene showed significant correlation with reduction of A antigen expression in TCC, while the expression of the A antigen is maintained in concomitant dysplasia or normal urothelium, suggesting that loss of the ABO gene and/or its promoter hypermethylation is a specific marker for TCC. PMID- 15880138 TI - Modulation of Ca(2+)-dependent and Ca(2+)-independent miniature endplate potentials by phorbol ester and adenosine in frog. AB - Phorbol esters and adenosine modulate transmitter release from frog motor nerves through actions at separate sites downstream of calcium entry. However, it is not known whether these agents have calcium-independent sites of action. We therefore characterised calcium independent miniature endplate potentials (mepps) generated in response to 4-aminoquinaldine (4-AQ(A)) and then compared the modulation of these mepps by phorbol esters and adenosine with that of normal calcium dependent mepps. Application of 30 microM 4-AQ(A) resulted in the appearance of a population of mepps with amplitudes greater than twice the total population mode (mepp(>2M)). In the presence of 4-AQ(A), K(+) depolarisation or hypertonicity increased the numbers of normal amplitude mepps (mepp(N)) but had no effect on the frequency of mepp(>2M) events, suggesting that mepp(>2M) are not dependent on calcium. Treatment with the botulinum toxin (Botx) fractions C, D, or E (which selectively cleave syntaxin, synaptobrevin and SNAP-25, respectively) produced equivalent reductions in both normal and 4-AQ(A) induced mepps, suggesting that both mepp populations have equal dependence on the intact SNARE proteins. Phorbol dibutyrate (PDBu, 100 nM) increased the frequencies of both populations of mepps recorded in the presence of 4-AQ(A). Adenosine (25 microM) selectively reduced the numbers of mepp(N) with no effect on the frequency of mepp(>2M) events. These results suggest that mepp(>2M) events released in response to 4-AQ(A) are dependent on intact forms of syntaxin, synaptobrevin and SNAP-25, but unlike mepp(N) are independent of a functional calcium sensor. The selective action of adenosine, to reduce the numbers of normal amplitude mepps without effecting the frequency of mepp(>2M) events, suggests that adenosine normally inhibits transmitter release through a mechanism that is dependent on the presence of a functional calcium sensor. PMID- 15880139 TI - Pomegranate flower improves cardiac lipid metabolism in a diabetic rat model: role of lowering circulating lipids. AB - Excess triglyceride (TG) accumulation and increased fatty acid (FA) oxidation in the diabetic heart contribute to cardiac dysfunction. Punica granatum flower (PGF) is a traditional antidiabetic medicine. Here, we investigated the effects and mechanisms of action of PGF extract on abnormal cardiac lipid metabolism both in vivo and in vitro. Long-term oral administration of PGF extract (500 mg kg( 1)) reduced cardiac TG content, accompanied by a decrease in plasma levels of TG and total cholesterol in Zucker diabetic fatty (ZDF) rats, indicating improvement by PGF extract of abnormal cardiac TG accumulation and hyperlipidemia in this diabetic model. Treatment of ZDF rats with PGF extract lowered plasma FA levels. Furthermore, the treatment suppressed cardiac overexpression of mRNAs encoding for FA transport protein, peroxisome proliferator-activated receptor (PPAR) alpha, carnitine palmitoyltransferase-1, acyl-CoA oxidase and 5'-AMP-activated protein kinase alpha2, and restored downregulated cardiac acetyl-CoA carboxylase mRNA expression in ZDF rats, whereas it showed little effect in Zucker lean rats. The results suggest that PGF extract inhibits increased cardiac FA uptake and oxidation in the diabetic condition. PGF extract and its component oleanolic acid enhanced PPAR-alpha luciferase reporter gene activity in human embryonic kidney 293 cells, and this effect was completely suppressed by a selective PPAR-alpha antagonist MK-886, consistent with the presence of PPAR-alpha activator activity in the extract and this component. Our findings suggest that PGF extract improves abnormal cardiac lipid metabolism in ZDF rats by activating PPAR-alpha and thereby lowering circulating lipid and inhibiting its cardiac uptake. PMID- 15880140 TI - Pharmacodynamics of propiverine and three of its main metabolites on detrusor contraction. AB - 1. Besides its antimuscarinic effects, propiverine may possess an additional mode of action. We compared the effects of propiverine, three of its metabolites (M-5, M-6, M-14) and atropine in human, pig and mouse urinary bladder preparations in order to elucidate the nature of a possible additional mode of action. 2. Like the parent compound, M-5, M-6 and M-14 reduced to variable degrees the contractions elicited by electric field stimulation (EFS) of isolated, urothelium denuded detrusor strips. In mouse the atropine-resistant and therefore the nonadrenergic, noncholinergic component of contractile response to EFS was reduced by M-5, M-14 and propiverine, but was hardly affected by M-6. 3. Atropine, propiverine and M-6 significantly shifted the cumulative concentration response curves for carbachol (CCh) to higher concentrations. Atropine and M-6 did not affect the maximum tension induced by CCh. Propiverine, M-5 and M-14 reduced the maximum CCh effect, suggesting at least one additional mode of action. This pattern of response was observed in all the three species, albeit with some differences in sensitivity to the various agents. 4. In freshly isolated human detrusor smooth muscle cells, propiverine and M-14 inhibited the nifedipine-sensitive L-type calcium current (I(Ca)) in a concentration-dependent manner. In contrast, the effects of M-5 and M-6 on I(Ca) were insignificant in the concentration range examined. 5. The investigated responses to propiverine and its metabolites suggest that impairment of maximum CCh-induced contractions is due to strong effect on I(Ca) and that this may be associated with the presence of the aliphatic side chain. PMID- 15880141 TI - Substrate-induced modulation of glutamate uptake in human platelets. AB - In the central nervous system (CNS), glutamate rapidly upregulates the activities of different excitatory amino-acid transporter subtypes (EAATs) in order to help protect neurons from excitotoxicity. Since human platelets display a specific sodium-dependent glutamate uptake activity, and express the three major glutamate transporters, which may be affected in neurological disorders, we investigated whether platelets are subject to substrate-induced modulation as described for CNS. A time- and dose-dependent upregulation of [3H]-glutamate uptake (up to two fold) was observed in platelets preincubated with glutamate. There was an increase in maximal velocity rate without affinity changes. Glutamate receptor agonists and antagonists did not modulate this upregulation and preincubation with glutamate analogues failed to mimic the glutamate effect. Only aspartate preincubation increased the uptake, albeit approximately 35% less with respect to glutamate. The effect of glutamate preincubation on the expression of the three major transporters was studied by Western blotting, showing an increase of approximately 70% in EAAT1 immunoreactivity that was completely blocked by cycloheximide (CEM). However, L-serine-O-sulphate, at a concentration (200 microM) known to block EAAT1/3 selectively, did not completely inhibit the effect of glutamate stimulation, indicating the possible involvement of EAAT2. In fact, glutamate stimulation was completely abolished only when, following CEM pre incubation, the experiment was run in the presence of the selective EAAT2 inhibitor dihydrokainic acid. Since surface biotinylation experiments failed to show evidence of EAAT2 translocation, our results suggest the existence of a different way of regulating EAAT2 activity. These findings indicate that human platelets display a substrate-dependent modulation of glutamate uptake mediated by different molecular mechanisms and confirm that ex vivo platelets are a reliable model to investigate the dysfunction of glutamate uptake regulation in patients affected by neurological disorders. PMID- 15880142 TI - Carbon monoxide-releasing molecules (CO-RMs) attenuate the inflammatory response elicited by lipopolysaccharide in RAW264.7 murine macrophages. AB - The enzyme heme oxygenase-1 (HO-1) is a cytoprotective and anti-inflammatory protein that degrades heme to produce biliverdin/bilirubin, ferrous iron and carbon monoxide (CO). The anti-inflammatory properties of HO-1 are related to inhibition of adhesion molecule expression and reduction of oxidative stress, while exogenous CO gas treatment decreases the production of inflammatory mediators such as cytokines and nitric oxide (NO). CO-releasing molecules (CO RMs) are a novel group of substances identified by our group that are capable of modulating physiological functions via the liberation of CO. We aimed in this study to examine the potential anti-inflammatory characteristics of CORM-2 and CORM-3 in an in vitro model of lipopolysaccharide (LPS)-stimulated murine macrophages. Stimulation of RAW264.7 macrophages with LPS resulted in increased expression of inducible NO synthase (iNOS) and production of nitrite. CORM-2 or CORM-3 (10-100 microM) reduced nitrite generation in a concentration-dependent manner but did not affect the protein levels of iNOS. CORM-3 also decreased nitrite levels when added 3 or 6 h after LPS exposure. CORM-2 or CORM-3 did not cause any evident cytotoxicity and produced an increase in HO-1 expression and heme oxygenase activity; this effect was completely prevented by the thiol donor N-acetylcysteine. CORM-3 also considerably reduced the levels of tumor necrosis factor-alpha, another mediator of the inflammatory response. The inhibitory effects of CORM-2 and CORM-3 were not observed when the inactive compounds, which do not release CO, were coincubated with LPS. These results indicate that CO liberated by CORM-2 and CORM-3 significantly suppresses the inflammatory response elicited by LPS in cultured macrophages and suggest that CO carriers can be used as an effective strategy to modulate inflammation. PMID- 15880143 TI - Effect of serum withdrawal on the contribution of L-type calcium channels (CaV1.2) to intracellular Ca2+ responses and chemotaxis in cultured human vascular smooth muscle cells. AB - Vascular smooth muscle cell (VSMC) chemotaxis is fundamental to atherosclerosis and intimal hyperplasia. An increase in intracellular Ca2+ [Ca2+]i is an important signal in chemotaxis, but the role of L-type calcium channels (CaV1.2) in this response in human vascular smooth muscle cells (hVSMC) has not been examined. hVSMC were grown from explant cultures of saphenous vein. Confluent hVSMC at passage 3 were studied after culture in medium containing 15% foetal calf serum (FCS) (randomly cycling) or following serum deprivation for up to 7 days. Smooth muscle alpha-actin was measured by immunoblotting and immunofluorescence microscopy. [Ca2+]i was measured using fura 2 fluorimetry. Chemotaxis was measured using a modified Boyden chamber technique and cell attachment to gelatin-coated plates was also quantified. The number and affinity of dihydropyridine-binding sites was assessed using [5-methyl-3H]PN 200-110 binding. In randomly cycling cells, the calcium channel agonist, Bay K 8644a and 100 mM KCl did not affect [Ca2+]i. In addition, the rise in [Ca2+]i induced by platelet-derived growth factor-BB (PDGF) was unaffected by the CaV1.2 antagonists, amlodipine and verapamil. In randomly cycling cells amlodipine did not affect PDGF-induced migration. In serum-deprived cells, smooth muscle alpha actin was increased and Bay K 8644a and 100 mM KCl increased [Ca2+]i. PDGF induced rises in [Ca2+]i were also inhibited by amlodipine and verapamil. The ability of Bay K 8644a to increase [Ca2+]i and verapamil to inhibit PDGF-induced rises in [Ca2+]i was evident within 3 days after serum withdrawal. In serum deprived hVSMC Bay K 8644a induced chemotaxis and amlodipine inhibited PDGF induced migration. Cell attachment in the presence of PDGF was unaffected by amlodipine in either randomly cycling or serum-deprived hVSMC. Serum withdrawal was associated with a decrease in the maximum number of dihydropyridine-binding sites (B(max)) and a decrease in affinity (K(D)). Serum deprivation of hVSMC results in increased expression of smooth muscle alpha-actin, a marker of more differentiated status, and increased [Ca2+]i responses and chemotaxis mediated by CaV1.2. These observations may have important implications for understanding the therapeutic benefits of calcium channel antagonists in cardiovascular disease. PMID- 15880144 TI - Effect of glutamate receptor antagonists on place aversion induced by naloxone in single-dose morphine-treated rats. AB - The neurobiological mechanism underlying the negative motivational component of withdrawal from acute opiate dependence is far from understood. Our objectives were to determine whether the glutamatergic system is involved in the motivational component of morphine withdrawal in acutely dependent rats and such an involvement is associated with dopaminergic neurotransmission. We examined the effects of various kinds of glutamate receptor antagonists on conditioned place aversion (CPA) induced by naloxone-precipitated withdrawal from a single morphine exposure 24 h before. Furthermore, the influence of pretreatment with the dopamine receptor antagonist haloperidol on those effects of glutamate receptor antagonists was also investigated. CPA was attenuated in a dose-dependent manner by all glutamate receptor antagonists examined including the NMDA receptor antagonists (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclo-hepten-5,10-imine maleate (MK-801) and phencyclidine hydrochloride (PCP), AMPA receptor antagonist 1-(4-aminophenyl)4-methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine hydrochloride (GYKI 52466), and metabotropic receptor antagonists (+/-)-2-amino-3 phosphonopropionic acid (AP-3) and (+/-)-alpha-methyl-4-carboxyphenylglycine (MCPG). The effects of MK-801, GYKI 52466 and MCPG were blocked by haloperidol. These results suggest that the glutamatergic system involving multiple classes of receptors plays a role in the motivational component of withdrawal from acute morphine dependence, and the function of the glutamatergic system would be closely associated with dopaminergic neurotransmission. PMID- 15880146 TI - [Dangerous delusions]. PMID- 15880145 TI - SO-3, a new O-superfamily conopeptide derived from Conus striatus, selectively inhibits N-type calcium currents in cultured hippocampal neurons. AB - Whole-cell currents in cultured hippocampal neurons were recorded to investigate the effects of SO-3, a new O-superfamily conopeptide derived from Conus striatus, on voltage-sensitive channels. SO-3 had no effect on voltage-sensitive sodium currents, delayed rectifier potassium currents, and transient outward potassium currents. Similar to the selective N-type calcium channel blocker omega-conotoxin MVIIA (MVIIA), SO-3 could concentration-dependently inhibit the high voltage activated (HVA) calcium currents (I(Ca)). MVIIA(3 microM), 10 microM nimodipine, and 0.5 microM omega-agatoxin IVA (Aga) could selectively block the N-, L-, and P/Q-type I(Ca), which contributed approximately 32, approximately 38, and approximately 21% of the HVA currents in hippocampal neurons, respectively. About 31% of the total HVA currents were inhibited by 3 microM SO-3. SO-3 (3 microM) and 3 microM MVIIA inhibited the overlapping components of HVA currents, whereas no overlapping component was inhibited by 3 microM SO-3 and 10 microM nimodipine, or by 3 microM SO-3 and 0.5 microM Aga. Also, 3 microM SO-3 had no effect on R type currents. SO-3 had less inhibitory effects on non-N-type HVA currents than MVIIA at higher concentrations (30 and 100 microM). The inhibitory effects of SO 3 and MVIIA on HVA currents were almost fully reversible. However, the recovery from block by MVIIA was more rapid than recovery from block by SO-3. It is concluded that SO-3 is a new omega-conotoxin selectively targeting N-type voltage sensitive calcium channels. Considering the significance of N-type calcium channels for pain transduction, SO-3 may have therapeutic potential as a novel analgesic agent. PMID- 15880147 TI - [Quality improvement in Norwegian hospitals]. PMID- 15880148 TI - [Indicators--beneficial, but problematic]. PMID- 15880149 TI - [Surgical treatment of carotid stenoses at a county hospital]. AB - BACKGROUND: High-degree stenosis of the internal carotid artery is a significant risk factor for cerebral infarction. It has been clearly demonstrated that endarterectomy reduces this risk if perioperative complications are maintained on a low level. MATERIAL AND METHODS: We studied the files of all patients undergoing carotid surgery in our hospital from 1985 through 2001. We also mailed questionnaires to all who were still alive in 2002. RESULTS: Over the period, 195 operations were performed on 168 patients (mean 12 operations per year). Their mean age was 63; 67% were men. In 30 cases (15%) the stenosis was asymptomatic. Within 30 days after surgery four patients had a disabling stroke, one had a retinal infarction and one patient died. The rate of perioperative stroke or death was 6.1%, while the rate for disabling stroke or death was 2.6%. Peripheral nerve injury, most often transitory, occurred in 5.7%. INTERPRETATION: The risk of moderate or serious perioperative complications is comparable to the multicentre studies that provide the rationale for the treatment (NASCET and ECST). Carotid endarterectomy in our hospital is safe in preventing stroke and should still be offered patients presenting to our centre. PMID- 15880150 TI - [Long term prognosis in relation to the presence of systolic heart murmurs in healthy middle-aged men]. AB - BACKGROUND: The long-term prognostic value of systolic murmurs revealed by heart auscultation has previously not been published. In this survey the prognostic value of systolic murmurs has been studied in relation to coronary heart disease and aortic valve operations. MATERIAL AND METHODS: During 1972-75, a cohort of 2014 apparently healthy men (40-59 years) from five companies in Oslo, Norway underwent heart auscultation under standardized conditions. Systolic murmurs were graded from I to VI. The men were prospectively followed up for 21.5 years in order to study the frequency of aortic valve operations, myocardial infarctions and coronary bypass operations. RESULTS: Modest systolic murmurs (grade I-II, n = 441) were associated with an unadjusted relative risk of 5.4 (95% CI 2.1-14.0), and moderate to strong murmurs (grade III-IV: n = 32) with a relative risk of 114.6 (95% CI 44.9-292.1) for aortic valve operation over the course of 21.5 years. The incidence of myocardial infarctions did not show any significant relationship to murmurs. Among those who underwent aortic valve surgery and who had a baseline murmur > or = III, a fourfold increase in bypass operations was observed. INTERPRETATION: Apparently healthy middle-aged men with systolic murmurs grade III or IV revealed by heart auscultation should be followed up carefully with regard to future need for aortic valve surgery. The increased frequency of coronary bypass operations among those with systolic murmur grade III or IV is possibly a result of aortic valve and bypass surgery being performed simultaneously. PMID- 15880151 TI - [Sources of intoxication--how do adolescents get hold of alcohol?]. AB - BACKGROUND: Norway has a legal age limit for buying beer and wine of 18, 20 for spirits. Knowledge about how under-aged adolescents procure alcohol and the relative importance of the various sources is important in order to reduce the availability of alcohol among adolescents. MATERIAL AND METHODS: In a national school-based survey among 12,000 students in junior and senior high school a sub sample comprising 13 to 17-year-old current drinkers were asked to state the frequency with which they had obtained alcohol from 8 different sources in the past 12 months. RESULTS: On average the respondents had obtained alcohol 11 times during the past year. The most frequently used source was "other adolescents" which constituted one third of the total number of times when alcohol had been obtained. Other important sources were "at home" (given or stolen), 20%; "off premise and on-premise sales" (stores, wine and spirits outlets, bars, pubs etc., 22%; and "other sources", 19%. Older adolescents and those who drank frequently obtained alcohol more often and from a larger number of sources than others. INTERPRETATION: Enforcing the legal age limits in on-premise and off-premise alcohol outlets as well as limiting adolescents' access to alcohol at home may reduce consumption and related harm among adolescents. PMID- 15880152 TI - [Hemoglobinopathies and patients with foreign names]. AB - BACKGROUND: The diagnosis of haemoglobinopathies is of growing importance in Norway because of increasing immigration from countries where haemoglobinopathies are prevalent conditions. The aim of this study was to investigate the relationship between mean corpuscular volume (MCV) and the various haemoglobinopathies diagnosed in Norway. MATERIAL AND METHODS: For a period of three years, all samples with MCV lower than 70 fl were also examined for beta thalassaemia and haemoglobin variants HbS, HbC, HbE and HbD. A total of 263 samples with low MCV were analysed by high-pressure liquid chromatography. RESULTS AND INTERPRETATION: In 18% of the samples, a variant of haemoglobinopathy was found, mainly beta-thalassemia minor. 119 of the samples were from persons with an ethnic background from a country in which these diseases are common; all observed haemoglobinopathies were found in this group. 35% of persons with low MCV and a mainly African or Asian ethnic origin had a heterozygous haeomglobinopathy. Low MCV in patients with a foreign ethnic origin is a useful first step in the diagnosis of haemoglobinopathies. PMID- 15880153 TI - [Infections in immunosuppressed children]. AB - BACKGROUND: Children undergoing transplantation or treatment for cancer have periods with severe immunosuppression; hence they are very susceptible to infections. A bacterial infection can rapidly become life threatening, and it is crucial to promptly start antibiotic treatment. MATERIALS AND METHODS: The background for this article is a two-day discussion among Norwegian paediatricians about infections in immunosuppressed children. In addition we have reviewed the literature by searches in PubMed, reference books and international guidelines. RESULTS AND INTERPRETATION: When a neutropenic patient becomes febrile, one should quickly do a thorough clinical examination, secure relevant microbiological samples, and start treatment with broad-spectrum antibiotics. As standard treatment for Norwegian children we recommend a combination of intravenous ampicillin and gentamicin. Patients who have clinical signs of septic shock should be given cefotaxime and gentamicin. If they get worse or show no signs of recovery after 3 to 5 days, a change to monotherapy with cefotaxime is recommended. Patients already treated with cefotaxime should be switched to meropenem, possibly in combination with vancomycin. Antifungal and/or anti anaerobic treatment should also be considered. PMID- 15880154 TI - [Teenage smoking and lung cancer incidence in early adult age, 1954-1998]. AB - OBJECTIVE: To describe national trends in Norway in lung cancer incidence among young adults and its relationship to adolescent smoking. MATERIAL AND METHODS: Between 1954 and 1998, a total of 1108 non-carcinoid lung cancers in individuals aged 20-44 were reported to the Cancer Registry of Norway. Temporal variations were studied in age and sex-specific rates, in age-adjusted rates, and in cohort specific rates. The association between cancer incidence and smoking prevalence was evaluated. RESULTS: The incidence among women aged 40-44 in Norway continued to increase into the most recent time interval (1994-1998) whereas the rate among men aged 40-44 was essentially constant after 1970. Consequently, the incidence rates converged among male and female young adults. The incidence at age 40-44 was highly correlated with smoking prevalence at age 15-19 in males (r = 0.88) and females (r = 0.82) within the same birth cohort. INTERPRETATION: The lung cancer incidence in young Norwegian women now equals that of men. The risk at age 40-44 was closely associated with teenage smoking, indicating that duration and age of onset are important. PMID- 15880155 TI - [Drug screening among patients aged 17-40 admitted with psychosis]. AB - BACKGROUND: High frequency of co-occurring substance use disorders and psychiatric disorders is reported in several studies. An important issue is whether we can rely on patient reports only. The purpose of this investigation was to find the prevalence of substance use among younger psychotic inpatients in Norway and to compare patients' self reports with the results of screening for drugs. MATERIAL AND METHODS: 65 patients aged 17 to 40, consecutively admitted to Blakstad Psychiatric Hospital in 2001 with psychosis were interviewed with regard to substance use 30 days prior to admittance. The Addiction Severity Index (EuropASI) was used for interviews. Blood and urine samples for drug analysis were taken at admittance. RESULTS: 54% of the patients reported having used one or more substances for intoxication during the month prior to admittance. 40% of the included patients had used illegal drugs. Laboratory analyses revealed use of illegal drugs, mostly cannabis and amphetamine, in 34% of the patients. Only one patient tested positively for a drug not reported in the interview. INTERPRETATION: The investigation shows that 54% of the younger psychotic inpatients intoxicate themselves with legal or illegal substances. This fact is important for the total medical management of these patients in psychiatric hospitals. We also found that this group of patients were reliable in reporting their substance use when a standardised interview was used. PMID- 15880156 TI - [Basic pharmacokinetics--elimination]. PMID- 15880157 TI - [Assessment of dementia and use of anti-dementia drugs in nursing homes]. AB - BACKGROUND: About 75% of nursing homes patients in Norway suffer from dementia, though only half of them have a dementia diagnosis. No consensus exists on how or when to do an assessment for dementia in the nursing home. MATERIAL AND METHODS: The paper is based on a search on Medline and personal experience in the field. RESULTS AND INTERPRETATION: Dementia assessment in nursing homes should be done in a co-operation between a physician, a nurse and an occupational therapist. The physician should carry out a Mini-Mental State Examination and a clock test, a physical and mental examination, an assessment of depressive symptomatology, and see to it that the thyroid function is measured. A CT scan of the brain should be done in cases with short duration of dementia symptomatology. A nurse and an occupational therapist should interview a family member and assess the patient's functioning by means of the Informant questionnaire on cognitive decline in the elderly and the Clinical dementia rating scale, or a similar assessment tool. Memantine could be a treatment for some patients with moderate to severe degrees of dementia, while acetylcholinesterase inhibitors could be useful for some patients with mild to moderate degrees of dementia of the Alzheimer type. Anti dementia drug therapy must be individualised. PMID- 15880158 TI - [Beneficial effects of dementia care units]. AB - BACKGROUND: Somatic nursing homes are the cornerstones of institutionalised care for people with dementia; the proportion of nursing home patients suffering from dementia is above 70%. Although most communities provide small dementia care units, they do not meet the needs and their aims are poorly defined. In this article, the effects of special care units on demented patients are examined. METHODS: Literature searches on Medline and Ageline were supplemented by reports from the Norwegian Centre for Dementia Research. RESULTS: The few studies identified indicate that small dementia care units are beneficial for patients, giving them a better quality of life. Patients maintain their cognition and functional ability longer, they are more relaxed, and receive less antipsychotic drugs. Various types of constraint and surveillance are used less often. The beneficial effects of special care units vary among patients, and they decrease over time. INTERPRETATION: Patients benefit temporarily from living in special dementia care units. However, the aims of these units are poorly defined and standards for staffing and the selection of patients are so far lacking. Further studies of quality and effect on care are needed, as the literature is scarce. PMID- 15880159 TI - [Physical training for nursing home residents--has it any effect?]. AB - BACKGROUND: The paper presents documentation of the effect of physical training for nursing home residents. MATERIAL AND METHODS: Literature study based on review articles and exercise trials in nursing home populations. RESULTS AND INTERPRETATION: The relatively few studies available indicate that physical training in a nursing home setting could be carried out. One study, reporting the effect of a fourfold increase in physiotherapy and occupational therapy, found positive effects on physical functioning and nursing costs. Furthermore, training is reported to improve aspects of physical functioning, like muscle strength and mobility. Several studies report effect on incontinence of low-dose physical exercises combined with scheduled toilet training. No study reports effect of physical training alone on the incidence of falls among nursing home residents. Nevertheless, physical training seems to improve physical fitness and functioning, cognition and behaviour. It is suggested that physical training for nursing home residents is carried out routinely in order to maintain physical functioning. Important aspects of such training are muscle strength, balance, coordination, endurance and flexibility. However, the most crucial type of training is active participation in the activities of daily life in order to prevent loss of independence. PMID- 15880160 TI - [Cooperation between municipalities and specialist health services--experiences from Trondheim]. PMID- 15880161 TI - [Mutual evaluation as a method of quality assurance in hospital units]. AB - BACKGROUND: Mutual visit schemes have been used in quality assurance of general practice. We have examined the effect of mutual evaluation in the quality assurance of patient care in hospital units. MATERIAL AND METHODS: Four university hospital units for medical gastroenterology received and performed one visit each. The visits were performed by one doctor and two nurses and lasted two days. The visit resulted in written reports that were first discussed locally and subsequently jointly in connection with a seminar. The evaluation was based on reported experiences, proposed improvements and surveys of patient satisfaction before and after the mutual evaluation process. A method was developed for surveying patient satisfaction with outpatient endoscopy. RESULTS AND INTERPRETATION: The mutual evaluation created enthusiasm, increased understanding and generated many proposals for improvement. Some improvement was observed in patients' satisfaction with the information given. The study suggests that mutual evaluation is a suitable method for quality assurance of patient care in hospital units. PMID- 15880162 TI - [Pharmacy-managed drug supply may reduce drug expenditure in hospitals]. AB - BACKGROUND: In order to reduce the cost of drugs and the time nurses spend on them, a six-month collaboration project was carried out between the hospital pharmacy and a ward unit. The ward unit had a yearly drug expenditure of 5.9 million NOK. MATERIALS AND METHODS: Pharmacy technicians stocked up the unit twice a week according to an agreed standard. Hospital pharmacists checked the need of other drugs ordered. A comparison between periods before and during the project was done on the value of the drug stock, credited returned drugs, nurses' time spent on drug handling and the number of telephone orders to the pharmacy. The value of avoided deliveries was calculated. RESULTS: The value of the drug stock was halved and calculated at a value of NOK 22,000. The value of drugs credited by the pharmacy was NOK 52,000. The savings from avoided deliveries were NOK 90,000. Total savings were NOK 164,000, 5% of the value of total purchases. In addition, nurses spent 11 hours less a week on drug handling. The number of telephone orders to the pharmacy was halved. INTERPRETATION: Pharmacy-managed drug supply to hospital wards may reduce drug expenditure and the risk of medication errors. PMID- 15880166 TI - [Fire in a physician's office--a warning]. PMID- 15880167 TI - [Diet and pain]. PMID- 15880168 TI - [Resource to medical research should be doubled]. PMID- 15880171 TI - [The Baerum issue-again]. PMID- 15880173 TI - [MR imaging in whiplash injuries]. PMID- 15880174 TI - [As love, almost]. PMID- 15880175 TI - [Stem cells--is it so important?]. PMID- 15880179 TI - The rising tide: combating coastal pollution. AB - Coastal zones are major hubs for economic and social activity and are in the front line of climate change. To safeguard these fragile ecosystems for our own and future generations we have to move towards more integrated approaches to the assessment and management of coastal and marine environments. Environmental monitoring-broadly defined-has a huge role to play. PMID- 15880180 TI - [Clinical features of Wegener granulomatosis and microscopic polyangiitis in Chilean patients]. AB - BACKGROUND: Systemic vasculitis are a group of heterogeneous diseases characterized by inflammation and necrosis of blood vessel walls. The etiology is not known, but geographic and environmental factors are implicated. AIM: To describe the clinical features of microscopic polyangiitis (MPA) and Wegener's granulomatosis (WG) in a Chilean cohort of patients. PATIENTS AND METHODS: Retrospective review of the medical records of 123 patients with the diagnosis of systemic vasculitis (65 MPA and 58 WG), seen from 1990 to 2001. The diagnosis were made based on the American College of Rheumatology and Chapel Hill criteria. RESULTS: The mean follow-up for MPA was 15 months (1-120) and for WG, 20 months (1-120). The median age (years) at diagnosis for MPA was 61 (19-82) and WG 50 (20 82). Gender distribution was similar in both groups (male: 68% and 57% respectively). The main clinical features in the MPA group were renal involvement (68%), peripheral nervous system involvement (57%), pulmonary hemorrhage (28%), and skin disease (32%). In the WG group were alveolar hemorrhage (62%), renal involvement (78%), paranasal sinus involvement (57%), and ocular disease (26%). In both, creatinine levels above 2.0 mg/dl were associated with a higher mortality (p< 0.01). ANCA by immunofluorescence was performed in 56 MPA patients (75% had pANCA, 4% had cANCA and 21% were ANCA negative) and in 55 WG patients (17% had pANCA, 79% had cANCA and 4% were ANCA negative). Global mortality was 18% and 17% respectively, and the most common causes of death were infections. CONCLUSIONS: The clinical features of our patients are similar to other published data. In our WG and MPA patients the main predictor for death was a serum creatinine above 2 mg/dl. PMID- 15880181 TI - [Valve repair surgery for incompetent bicuspid aortic valves]. AB - BACKGROUND: Surgical valve repair is a good alternative for correction of incompetent bicuspid aortic valve. AIM: To report the early and late surgical, clinical and ecochardiographic results of surgical repair of incompetent bicuspid aortic valves. PATIENTS AND METHODS: Retrospective review of medical records of 18 patients aged 19 to 61 years, with incompetent bicuspid aortic valve in whom a valve repair was performed. Four patients had infectious endocarditis and 17 were in functional class I or II. Follow up ranged from 3 to 113 months after surgery. RESULTS: A triangular resection of the prolapsing larger cusp, which included the middle raphe, was performed in 17 cases; in 13 of these, a complementary subcommisural annuloplasty was performed. In the remaining case, with a perforation of the non-coronary cusp, a pericardial patch was implanted; this procedure was also performed in 2 other cases. In 3 cases large vegetations were removed. Postoperative transesophageal echocardiography showed no regurgitation in 11 patients (62%) and mild regurgitation in 7 (38%). There was no operative morbidity or mortality. There were no deaths during the follow-up period. In 3 patients (17%) the aortic valve was replaced with a mechanical prosthesis, 8 to 108 months after the first operation. Reoperation was not needed in 93%+/-6,4% at 1 year and 85%+/-9,5% at 5 years, these patients were all in functional class I at the end of the follow-up period. 60% had no aortic regurgitation, 20% had mild and 20% moderate aortic regurgitation on echocardiographic examination. A significant reduction of the diastolic diameter of the left ventricle was observed, but there were no significant changes in systolic diameter or shortening fraction. CONCLUSIONS: Surgical repair of incompetent bicuspid aortic valves has low operative morbidity and mortality and has a low risk of reoperation. PMID- 15880182 TI - [Antiphospholipid antibodies in idiopathic membranous nephropathy]. AB - BACKGROUND: Antiphospholipid antibodies have been found in the sera from patients with idiopathic and secondary glomerulopathies, mainly related to lupus. No special attention has been devoted to idiopathic membranous nephropathy, a glomerular disease with a high frequency of thrombotic complications, particularly of the renal vein. AIM: To study the presence and significance of antiphospholipid antibodies in idiopathic membranous nephropathy. MATERIAL AND METHODS: Anticardiolipin and anti-ss2-glycoprotein-I IgG antibodies were measured in serum samples from 21 patients with idiopathic membranous nephropathy (age range 11-75 years, 5 female). The medical records of 20 of these patients were reviewed, looking for vascular complications and nephrological evolution during a follow-up period that ranged from two to 277 months. RESULTS: Five patients had anticardiolipin antibody titers over the cutoff for normal values, and two others were positive for anti-ss2-glycoprotein-I, without cross-reactivity. There was no difference in the incidence of thrombotic complications in the renal vein, or other locations, between these seven patients and the remaining patients. No differences in the clinical course of the nephropathy were detected either. CONCLUSIONS: Antiphospholipid antibodies may be found in patients with primary membranous nephropathy. They are not related to thrombosis or a worse evolution. PMID- 15880183 TI - [Primary resistance to antiretroviral therapy in patients with HIV/AIDS in Chile]. AB - BACKGROUND: Resistance to antiretroviral therapy is a determining factor for therapeutic failure in HIV/AIDS. The prevalence of primary resistance (i.e. in those patients that have not received treatment) varies in different parts of the world. AIM: To study the prevalence of primary resistance to antiretroviral drugs in patients living in Northern Santiago. PATIENTS AND METHODS: Viral load, lymphocyte subpopulations by flow cytometry and genotypic resistance testing were assessed in blood samples from 60 HIV-1 infected patients (mean age 37 years, 54 male). RESULTS: Mean CD4 cell count and viral load was 200 cells/ml and 142,840 RNA copies/ml respectively. Ten mutations were identified: V179D, L10I/V, M361, L63P, A71T/V, Y115F, V118I and K20R. None of these mutations is associated to a high degree of resistance to reverse transcriptase inhibitors, nucleoside analogs (NRTI), non nucleoside analogs (NNRTI) or viral protease inhibitors. CONCLUSIONS: This is a first approach to study antiretroviral resistance in Chilean patients. This study must be amplified, since the prevalence of resistance may experience changes with time. PMID- 15880184 TI - [Seroprevalence of herpes simplex virus type 2 (HSV-2) infection in two clinics for sexually transmitted diseases in Santiago, Chile]. AB - BACKGROUND: Genital herpes, a sexually transmitted disease caused by herpes simplex virus type 2 (HSV-2), is found in 3.8% of all sexually transmitted diseases (STD) in Chile. AIM: To determine the seroprevalence of herpes simplex virus type 2 (HSV-2) and to identify an association between HSV-2 and HIV-1, other STD, and to study its demographic characteristics. MATERIAL AND METHODS: A cross-sectional study at two STD public clinics in Santiago was conducted among 200 consecutive patients. Samples were tested for HSV-2, HIV-1, syphilis and hepatitis B virus surface antigen. RESULTS: The seroprevalence for HSV-2 was 43%. Four patients had a history of genital herpes. There was a strong association between HSV-2 infection and HIV-1 positivity (OR=8.7, 95% CI 3.4-22.4, p <0.001) and the condition of being a sexual worker (OR=4.5, 95% CI 1.7-11.9, p <0.01). CONCLUSIONS: The high association with HIV-1 and sexual workers, emphasizes the need of having HSV-2 specific diagnostic tests, counseling on sign and symptom recognition and taking preventive measures. PMID- 15880185 TI - [Psychological factors associated to patient's treatment compliance in Chilean diabetic teenagers]. AB - BACKGROUND: Treatment compliance among patients with type 1 diabetes mellitus, is low in 50% of diabetic teenagers, becoming a social and medical problem. AIM: To determine psycho-social factors associated to treatment compliance among Chilean diabetic type 1 teenagers. PATIENTS AND METHODS: A non experimental study of 61 diabetic teenagers (age 14.9+/-1.9 years, 37 male). The number of blood glucose determinations, socioeconomic level and practice of sports was measured. Psychological tests were applied to analyze self-efficiency, motivation of achievement, self-esteem and knowledge of the illness and its treatment. As a measure of patient compliance, glycosilated hemoglobin (HB1Ac) was measured. RESULTS: Six patients had a good control of diabetes (HB1Ac <7%), 24 had HB1Ac values between 7 and 8.9, and 31 (51%) had values of 9% or more, considered as a poor diabetes control. The intensified insulin treatment scheme, the knowledge of the illness and its treatment and the sense of self-efficiency, were the factors associated with a better compliance with treatment. Teenagers of higher socio economical levels had a better compliance with treatment. CONCLUSIONS: Fifty percent of Chilean diabetic teenagers in this sample had a poor control of the disease and the variable knowledge about the disease is the better predictor of patient compliance. PMID- 15880186 TI - [Job performance and climacteric in female workers]. AB - BACKGROUND: During climacteric, the presence of disabling symptoms and the higher incidence of chronic diseases, may impair the job performance of women. AIM: To relate job performance levels with the climacteric period and associated factors in working women aged from 42 to 55 years old. SUBJECTS AND METHODS: In a cross sectional and correlative design, 64 secretaries at two public organizations at the Eighth Region of Chile, were assessed using the following instruments: Menopause-Specific Quality of Life Questionnaire (University of Toronto), adapted for Chile by PROSAM, Climacteric Self-care Questionnaire, Perceived Social Support Scale and Biodemographics Variables Questionnaire. A Perception of the Job Performance Scale was applied to the persons that superintended these woman. RESULTS: The Department Heads or persons supervising these women, determined that most workers did their work well. The best evaluations were given by older bosses. Most women under study showed a moderate alteration of the Menopause Quality of Life. A deficit of self-care during the climacteric period was detected in 92.2%. The perceived social support from friends correlated with job performance. Menopause quality of life in its different domains had no correlation with job performance variable. CONCLUSIONS: No association between changes during the climacteric period and job performance was observed among women participating in this study. PMID- 15880187 TI - [Thyroid microcarcinoma with an aggressive evolution. Report of one case]. AB - The treatment of papillary thyroid carcinoma of less than 10 mm diameter is a matter of controversy. The incidental finding of papillary microcarcinomas in autopsies is frequent and some authors postulate that these tumors are biologically inactive and should only be observed. We report a 21 years old woman with a papillary thyroid cancer of 6 x 5 x 5 mm and bilateral paratracheal metastases, that was subjected to a total thyroidectomy. She received 200 mCi of radioiodine. Two years after surgery, a new nodule of 9.6 mm diameter was detected by ultrasound, that was treated with a new dose of 200 mCi of radioiodine. One year later a suprasternal mass of 2 cm diameter and 3 enlarged lymph nodes were detected. She was subjected to a surgical lymph node dissection of the neck and the biopsy confirmed the presence of cancer. She received a new dose of 300 mCi of radioiodine. The mother of the patient had a 7 mm thyroid nodule that was also a papillary carcinoma. PMID- 15880188 TI - [Atriocaval shunt. Report of two cases]. AB - The mortality of grade V and VI liver trauma fluctuates between 30% and 70%. The atriocaval shunt, described by Shrock et al, in 1968, is a therapeutic option that, after being installed, allows to repair the suprahepatic veins and retrohepatic cava in a bloodless surgical field. Its use requires an experienced and skilled surgeon to obtain survival rates similar to those obtained with other methods. We report two male patients of 17 and 18 years old treated successfully with this technique after suffering a blunt and a penetrating liver trauma by a shotgun, respectively. PMID- 15880189 TI - [Results of the INTRA-WHO survey in Chile]. AB - BACKGROUND: The aging speed in developing countries has been faster than predicted. Thus, health care systems must adapt to face this new scenario efficiently. The WHO designed the INTRA study to assess health promotion and protection actions in primary care, for people over 50 years of age. MATERIAL AND METHODS: Questionnaires designed by WHO were applied to 1,167 subjects (aged 50 94 years, 68% female) and 117 health care professionals attending and working respectively, at 33 health care centers of the Vina-Quillota Health Service. RESULTS: Twenty percent of subjects were illiterate and 25% had less than 6 years of instruction. Forty three percent could reach the health centre by public transportation and 92% did not need to be accompanied, 39% spent more than one hour to be attended and 71% considered that the service in the centre was good. Sixty seven percent attended regular appointments, 63% did not perform any physical activity and only half of them were advised to start such activity. Weight loss was recommended to 55% but only one third has achieved such goal. Only one third of patients admitted being interrogated about their drinking habits. Among subjects in whom blood pressure was measured, one fourth had abnormal values. CONCLUSIONS: Primary health care in Chile, although having health care programs for the elderly, is loosing opportunities to improve health status and quality of life of this age group. PMID- 15880190 TI - [Chronic kidney disease: classification, mechanisms of progression and strategies for renoprotection]. AB - Chronic kidney disease is a worldwide health problem. The incidence and prevalence of kidney failure is in constant increase, involving poor outcomes and high costs. The leading causes of kidney failure are type 2 diabetes and hypertension. The new initiative "Kidney Disease Improving Global Outcomes (KDIGO)" is a global public health approach to face this problem. A formal definition for chronic kidney disease and a staging of kidney diseases from kidney damage with preserved function to kidney failure, were proposed. We reviewed the main mechanisms involved in renal disease progression, with emphasis in the proteinuria and the intrarenal activation of renin angiotensin system. Moreover, the evidence in the literature of therapeutical interventions with proved efficacy in slowing the rate of reduction of renal function is discussed, particularly the optimal control of hypertension, reduction of proteinuria and renin angiotensin system blocking. Finally, we recommend a strategy for the clinical management of patients in the different stages of chronic kidney disease. PMID- 15880191 TI - [Genetic epidemiology of obesity: family studies]. AB - This review focuses on methodological aspects and main results of different family studies that have been conducted to assess the existence of a genetic contribution in human obesity. A genetic component in the etiology of obesity has been elucidated through specific study designs answering different research questions such as: a) Do obesity aggregate in families? b) Is there a genetic contribution to familial clustering? c) Is it possible to localize chromosomal regions that contain susceptibility genes to obesity? d) Is it possible to estimate the risk for developing obesity depending on the genotype profile in candidate genes? There are sufficient evidences indicating the existence of a moderate familial clustering of obesity defined as body mass index >/=30 with a stronger aggregation with more extreme values of body mass index. Twin studies have demonstrated that the familial aggregation of obesity has a genetic component and is not only due to cultural or environmental factors clustered in families. Linkage studies have identified markers and genes related to obesity in virtually all human chromosomes. However, some of these linkage studies have produced conflicting results. Discordant results are even more pronounced in case control studies that evaluate the association between alleles at candidate genes and obesity. Topics related to study design will acquire increasing importance in order to avoid methodological problems related to trait definition, sample sizes, population stratification by ethnicity and other confounding factors. PMID- 15880192 TI - [Homeostatic range of the oxidative metabolism: 60 years of integrative fisiometry]. AB - The energetic metabolism and its relationship with body weight generated a vivid controversy, since the Rubner's surface law was introduced into biology. Recently, the multifactor theory (Darveau et al) has caused again a revival of this polemic topic. Moreover, the investigations concerning metabolism and body weight include all terrestrial mammals, from the shrew (3 grams) to the elephant (three tons). The corresponding allometric exponent for standard metabolic rate, both theoretical and empirical, fluctuates around an average value of 0.75, in contrast with the surface law, which postulated a value of 0.67. The "metabolic range" (rest vs maximal exercise) does vary from 1 to 10, due to the prevalent influence of the skeletal muscle activity. Recent investigations have emphasized the fact that the allometric exponent is not unique (0.75), but it should be subjected to statistical variability, both in standard and in maximal exercise. PMID- 15880193 TI - [Notes on euthanasia]. AB - In the Judeo-Christian tradition, human life is held to be sacred, a semblance of the divine and a gift from God which the individual cannot dispose of at his or her own will. Hence, these monotheistic religions have made of the crime of murder a transgression of God's own commandment not to kill and have extended the applicability of this commandment to the practice of euthanasia and suicide. On the other hand, some non-religious traditions offer plausible reasons favoring euthanasia. This is a delicate matter for physicians, since the Hippocratic tradition forbids euthanasia and because as care-givers they must also bear the psychological, moral and emotional burden of carrying it out. Physicians are trained to preserve life but not to bring it to an end. As human beings, they must always respect the principle of nonmaleficence, and as physicians they must always respect as well the principle of beneficence. It is difficult to accept the fact that ending a human life can be an act of beneficence. In order to differentiate between passive and active euthanasia, the concept of proportionality of medical acts must be brought into consideration. For instance, using high doses of opiates to alleviate pain or withholding the use of an extraordinary method of treatment are not passive acts aimed at ending the life of a terminally ill patient, but medical acts that are reasonable, judicious and proportionate to the condition and irreversibility of a patient's illness. Therefore, so-called passive euthanasia cannot be considered the same as euthanasia. On the other hand, medically assisted suicide is a deceitful form of active euthanasia. The aim of this act is to cause death and the physician is morally responsible for such a death, since he is providing the means for bringing a human life to an end. Many times the desire to die expressed by terminally ill elderly and helpless patients is a request for help and an expression of reproach against a society that allows for their abandonment and neglect. PMID- 15880194 TI - [Seed, youth, the land is fertile and the moment, unique!: it is not Neruda, but Gandulfo, the surgeon]. AB - The second and definitive edition of "Crepusculario", the first book of Pablo Neruda, is dedicated by the poet "To Juan Gandulfo, this book of other times, Pablo". Juan Gandulfo, died tragically, at a very young age, in 1931. Juan Gandulfo, remembered in Neruda's memoirs, as the most formidable leader of the Anarchist movement of that time and that engraved in wood the cover and all the illustrations of the first edition of Crepusculario, published in 1923. Neruda qualified these as the "most impressive engravings made by a man that nobody relates to artistic creation". Juan Gandulfo, as a medical student, made the color illustrations used to teach biology and histology by Professor Juan Noe. Juan Gandulfo wrote more than one hundred articles in the magazine "Claridad", pertaining to the Student Federation of Chile. Juan Gandulfo was consequent with his writings and went to jail twice as a student leader and fought a duel for a woman. Later on, with the same vehemence, he dedicated himself to the care of children in the Arriaran Hospital and to teach general surgery at the San Vicente Hospital, but fate truncated his career at the age of 36. This essay is an homage to this man, in the centennial of Pablo Neruda's birth. PMID- 15880195 TI - [On the relevance of p values in medicine]. PMID- 15880196 TI - [Delirium from nicotine withdrawal in a post-operative adult patient]. PMID- 15880198 TI - [The prognosis of heart failure in Brazil: the search for reliable and representative data]. PMID- 15880199 TI - [Adrenergic cardiomyopathy: can stress cause acute heart disease?]. PMID- 15880200 TI - [Evaluation of segmentary contractility in Chagas' disease by using the integral of the myocardial velocity gradient (myocardial strain) obtained through tissue Doppler echocardiography]. AB - OBJECTIVE: To quantify the percentage of contractility of different myocardial segments in patients with Chagas' disease by measuring myocardial strain and to assess the differences in the radial and longitudinal ventricular contractile function in the undetermined and dilated forms of chronic chagasic cardiomyopathy as compared with those in a group of healthy individuals. METHODS: The study comprised 39 individuals [20 (51.3%) of the male sex] divided into the following 4 groups: 1) Nl: 17 (43.6%) healthy individuals; 2) Und: 7 (17.9%) patients with the undetermined form of Chagas' disease; 3) C1: 7 (17.9%) patients with the chronic form of Chagas' disease with ejection fraction < 50%; and 4) C2: 8 (20.5%) patients with the chronic form of Chagas' disease with ejection fraction > 50%. After performing baseline echocardiography, Doppler tissue images were recorded to measure myocardial strain in different segments on longitudinal and transversal parasternal, and apical 2- and 4-chamber views. RESULTS AND CONCLUSION: The percentage of contractility in the different myocardial segments, both the radial and longitudinal components, is greater in healthy individuals than in patients with the chronic form of Chagas' disease, and in those with the undetermined form of the disease as compared with that of chronic chagasic patients with EF < 50%. Left ventricular radial contractility is greater than left ventricular longitudinal contractility in all groups (Nl, Und, and Chronic). The data presented allow us to propose a progressive character of myocardial impairment in patients with Chagas' disease. PMID- 15880201 TI - [Prospective study of early and late morbidity and mortality in the abdominal aortic aneurysm surgical repair]. AB - OBJECTIVE: To prospectively assess early and late morbidity and mortality of patients undergoing elective surgical repair of abdominal infrarenal aortic aneurysms and to determine the independent predictors of cardiac events. METHODS: For 6 consecutive years, this study analyzed 130 patients, who underwent routine standardized preoperative assessment always with the same clinical, surgical, and anesthesia teams. RESULTS: In-hospital mortality was 3.1% (4 patients), and the major cause of death was mesenteric ischemia, which occurred in 3 patients. Forty eight (37%) nonsurgical complications occurred as follows: 8.5% were cardiac complications and 28.5% were noncardiac complications. The most common complications were the pulmonary ones, which occurred in 14 (10.8%) patients. Survivals in the first, third, and sixth postoperative years were 95%, 87%, and 76%, respectively. The variables that significantly correlated with morbidity and mortality were clinical predictors, mean age of 70.5 years, and presence of heart failure and chronic renal failure. No predictor of late morbidity and mortality was identified. CONCLUSION: Although this is considered a highly complex surgery, mortality is low, the cardiac complications are not very significant, and the patients have a good long-term evolution. PMID- 15880202 TI - [Correction of thoracic and thoracoabdominal aortic aneurysms. Central cannulation technique]. AB - OBJECTIVE: To demonstrate the viability of the use of extracorporeal circulation established between the left atrium and ascending aorta to induce deep hypothermia for correcting thoracic and thoracoabdominal aortic aneurysms. METHODS: From January 1994 to July 2001, 38 patients (mean age, 54.6 +/- 12.7 years) were operated on as follows: 12 (31.6%) patients underwent repair of thoracoabdominal aortic aneurysm, and 26 patients underwent repair of descending thoracic aneurysms. Deep hypothermia was induced by use of extracorporeal circulation, with pharyngeal temperature ranging from 15 to 25 degrees C (20.6 +/ 3.2 degrees C). RESULTS: Of the neurological complications, paraplegia occurred in 2 (5.3%) patients. One patient developed paraparesis, and another evolved with convulsion. Twelve (31.6%) patients had respiratory complications, and 2 (16.7%) patients died. Two other patients were operated on on an emergency basis and ended up dying. Total mortality was 18.4% (7 patients). CONCLUSION: Repair of descending thoracic and thoracoabdominal aortic aneurysms with deep hypothermia by use of extracorporeal circulation established between the left atrium and ascending aorta proved to be a viable method for correcting those aneurysms. PMID- 15880203 TI - [Blockade of renin-angiotensin system attenuates cardiac remodeling in rats undergoing aortic stenosis]. AB - OBJECTIVE: To assess the role of the AT1 receptor blocker and the angiotensin converting enzyme inhibitor in cardiac remodeling induced by aortic stenosis in rats. METHODS: Wistar rats were divided into the following 4 groups: 1) C- control (n=13); 2) AoS--aortic stenosis (n=11); 3) LIS--AoS treated with lisinopril, 20 mg/kg/day (n=11); and 4) LOS--AoS treated with losartan, 40 mg/kg/day (n=9). The treatments were initiated 3 days before surgery. After 6 weeks, the animals underwent echocardiographic study, and quantification of the hydroxyproline (HOP) concentration and the left ventricular (LV) myocyte cross sectional area (CSA). RESULTS: Aortic stenosis induced an increase in left ventricular wall thickness. The LIS and LOS groups showed no difference as compared with the control group. The AoS and LIS rats had greater left atrial diameters than the control rats did, while no difference was observed in the LOS animals. The AoS animals had greater values of shortening percentage than control animals did. This fact was modified with neither LIS nor LOS. The cross-sectional area of the animals in the AoS group was greater than that in the control group. However, treatment with LOS and LIS attenuated the AoS-induced increase in area. Aortic stenosis caused an increase in HOP concentration, while the LOS group showed no difference as compared with the control group. CONCLUSION: Blockade of the renin-angiotensin system with AT1 blocker and ACEI may attenuate the development of heart hypertrophy, but only the blockade of AT1 receptors attenuates left ventricular interstitial fibrosis. PMID- 15880204 TI - [Survival and prognostic factors in systolic heart failure with recent symptom onset]. AB - OBJECTIVE: To study survival and prognostic factors associated with mortality in patients with systolic heart failure followed up since symptom onset. METHODS: We carried out a study with a cohort of 204 consecutive patients with systolic heart failure, whose symptom onset occurred within the 6 weeks preceding the first medical visit. They were followed up for 46 months. The prognostic variables analyzed were collected when the patients were included in the study and were correlated with cardiovascular mortality. An EF < or =40% on echocardiography characterized systolic ventricular dysfunction. RESULTS: The overall survival rates according to the Kaplan-Meier technique were 98.0%, 90.6%, and 70.2% at 3, 12, and 48 months of follow-up, respectively. The multivariate analysis identified the independent effect of 6 variables on the risk of cardiovascular death. Functional classes III and IV increased risk 2.7 times as compared with class II; 10-mmHg increments in systolic blood pressure reduced the risk of death by 25%; each 10-bpm increase in heart rate increased the risk of death 1.6 times; and each 0.25-mg/dL increment in serum creatinine caused a 60% increase in risk. The presence of the third cardiac sound caused a 3-fold increase in the risk of death, and chagasic etiology was also associated with cardiovascular mortality (P<0.0001). CONCLUSION: Evidence shows that mortality in the initial phase is not elevated, and that etiology, advanced functional class, arterial hypotension, tachycardia, presence of the third cardiac sound, and elevated serum creatinine lead to a worse prognosis. PMID- 15880205 TI - [Effects of atorvastatin, fluvastatin, pravastatin, and simvastatin on endothelial function, lipid peroxidation, and aortic atherosclerosis in hypercholesterolemic rabbits]. AB - OBJECTIVE: To compare the effects of atorvastatin, fluvastatin, pravastatin, and simvastatin on endothelial function, aortic atherosclerosis, and the content of malondialdehyde (MDA) in native and oxidized LDL and in the arterial wall of hypercholesterolemic rabbits after adjusting the dosages of those statins to reduce total serum cholesterol levels to similar values. METHODS: Male rabbits were divided into the following 6 groups of 10 animals (n=10): 1) GH (control)- hypercholesterolemic animals; 2) GA--atorvastatin; 3) GF--fluvastatin; 4) GP- pravastatin; 5) GS--simvastatin; and 6) GN--normal. The animals were fed a standard food preparation enriched with 0.5% cholesterol and 2% coconut oil for 45 days. Fifteen days after beginning the experiment, atorvastatin, fluvastatin, pravastatin and simvastatin were administered for 15 days through gavage, and the dosages were adjusted to obtain similar cholesterol values in each group. At the end of the experiment, a blood sample was withdrawn for determining total cholesterol and separating the lipoproteins, and a segment of the thoracic aorta was removed to be used for studying endothelial function and lipid peroxidation, and for measuring aortic atherosclerosis in histological sections. RESULTS: The statins significantly reduced total serum cholesterol levels, LDL-cholesterol levels, and aortic atherosclerosis. The MDA content was also significantly reduced in native and oxidized LDL, as well as in the arterial wall. Endothelium dependent relaxation was significantly greater in the treated group compared with that in the hypercholesterolemic group. CONCLUSION: The statins, at dosages adjusted, had a significant and similar effect in reducing lipid peroxidation in native and oxidized LDL-C and in arterial walls, in decreasing aortic atherosclerosis, and in reverting endothelial dysfunction. PMID- 15880206 TI - [Chronic cigarette smoke exposure results in cardiac remodeling and impaired ventricular function in rats]. AB - OBJECTIVE: To determine the cardiac structural and functional alterations caused by cigarette smoke exposure in rats. METHODS: The animals were randomly distributed into the following 2 groups: 1) smokers (S), comprising 10 animals exposed to cigarette smoke at a rate of 40 cigarettes/day; and 2) control (C), comprising 10 animals not exposed to cigarette smoke. After 4 months, the animals underwent morphological and functional study with echocardiography. The variables studied were analyzed by use of the t test or the Mann-Whitney test. RESULTS: The smoking rats had a greater left atrium (S=4.2+/-0.7 mm; C=3.5+/-0.6 mm; P<0.05), and greater left ventricular diastolic (S=7.9+/-0.7 mm; C=7.2+/-0.5 mm; P<0.05) and systolic (S=4.1+/-0.5; C=3.4+/-0.5; P<0.05) diameters. The left ventricular mass index was greater in the smoking animals (S=1.5 mg/kg+/-0.2; C=1.3 mg/kg+/ 0.2; P<0.05), and the ejection fraction (S=0.85+/-0.03; C=0.89+/-0.03; P<0.05) and the shortening fraction (S=47.8%+/-3.7; C=52.7%+/-4.6; P<0.05) were greater in the control group. No differences were observed in the diastolic transmitral flow variables (E wave, A wave, and E/A ratio). CONCLUSION: Chronic cigarette smoke exposure results in cardiac remodeling with a decrease in ventricular functional capacity. PMID- 15880207 TI - [Nutritional status and lipid profile of postmenopausal women with coronary heart disease]. AB - OBJECTIVE: To assess the nutritional status and lipid profile of postmenopausal women with coronary heart disease. METHODS: A retrospective cross-sectional study was conducted with information obtained from the medical records of 217 women on the occasion of their first visit to the InCor Nutrition Outpatient Care Clinic. The data referred to the patients' nutritional status, body mass index, use of lipid-lowering medication, and serum lipid levels (cholesterol and fractions). RESULTS: The mean age was 60.98 +/- 9.23 years, and obesity prevailed in 56% of the patients. The use of lipid-lowering drugs was observed in 73% of the population. In regard to lipid profile, 56% had adequate serum levels of HDL-C. The nutritional status was inadequate due to the prevalence of obesity, which results in the appearance of other chronic diseases, such as dyslipidemias. Although the dosages of the lipid-lowering drugs used were not assessed, their use by the population studied did not seem to be favorable, because high levels of total cholesterol and LDL-C were observed, and those high levels in that condition are strongly related to the occurrence of cardiovascular diseases. CONCLUSION: Multidisciplinary action in the form of programs to promote women's health is required, comprising preventive aspects related to coronary heart disease, to improve the quality of life in that population. PMID- 15880208 TI - [Myotonic dystrophy and heart disease: behavior of arrhythmic events and conduction disturbances]. AB - OBJECTIVE: To study the prevalence and natural evolution of arrhythmic events and conduction disturbances in myotonic dystrophy; to correlate the genetic defect with cardiovascular findings; to assess cardiac mortality, frequency, and predictive factors of sudden death; to correlate the severity of the neuromuscular and cardiac involvement; and to define the role of the electrophysiological study (EPS), in myotonic dystrophy. METHODS: Periodic clinical assessment and the following tests were performed in 83 consecutive patients with a mean follow-up of 42+/-30.63 months: complementary examinations, genetic tests, electrocardiography, echocardiography, and Holter; electrophysiological study was performed in 59 cases. RESULTS: Atrial tachyarrhythmia was observed in 10 (12%) patients, NSVT in 14 (17%), first-degree AVB in 24 (29%), LBBB in 19 (23%), and RBBB in 13 (16%). Symptoms, an increase in the PR interval, QRS enlargement, LVEF < 60%, and age were predictive factors of death. Nine patients died (4 sudden deaths; 2 due to heart failure; 3 due to other causes). Electrophysiological study: H-V interval > 70 ms in 34% and > 100 ms in 11% (postprocainamide). CONCLUSION: The prevalence of arrhythmic events and conduction disturbances ranged from 50% to 80% after 6 years, and did not correlate with the genetic defect. Atrial flutter was the most common sustained arrhythmia. Cardiac involvement increased as the neuromuscular disease became aggravated, but progression of the cardiac involvement was more rapid than that of the neuromuscular disease. Overall mortality was low (11%) and sudden death occurred in half of the cases. The EPS identified a group at risk for pacemaker implantation. PMID- 15880209 TI - [Percutaneous transluminal coronary angioplasty in a patient with idiopathic thrombocytopenic purpura]. AB - The association between coronary heart disease and thrombocytopenic purpura is rare and poses some difficulties when myocardial revascularization is necessary. We report a case with this association and significant coronary impairment, which was percutaneously treated with stent implantation. PMID- 15880210 TI - [Transient ventricular dysfunction (Takotsubo cardiomyopathy)]. AB - The patient was a male with myasthenia gravis, hospitalized with acute respiratory failure due to decompensation of the underlying disease. He evolved with findings suggestive of acute myocardial infarction, with electrocardiographic and enzymatic alterations compatible with that diagnosis. The patient underwent emergency coronary angiography, which showed no severe coronary obstruction, although his left ventricle had significant systolic dysfunction with characteristic alterations, on ventriculography, of the syndrome described as transient ventricular dysfunction or Takotsubo syndrome. On evolution, complete recovery of the electrocardiographic alterations and systolic ventricular function assessed on echocardiography occurred, confirming the syndrome. PMID- 15880211 TI - [Intramural haematoma of the descending aorta]. PMID- 15880212 TI - [Circulatory shock in a 103 year-old man with a history of long lasting heart failure following acute myocardial infarction ten years before]. PMID- 15880213 TI - [Four-month-old infant with ventricular septal defect]. PMID- 15880214 TI - [Determination of hemodynamic parameters using Doppler two-dimensional echocardiography: a searching tool for therapeutic optimization in patients with congestive heart failure on an outpatient care follow-up]. PMID- 15880215 TI - Hepatitis C in patients co-infected with human immunodeficiency virus. A review and experience of a Brazilian ambulatory. AB - Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) share the same transmission mechanisms. The prevalence of HCV in the HIV-infected population varies from region to region, throughout the world, depending on different exposure factors to both viruses. Co-infection with HIV accelerates the progression of the disease caused by HCV, appears to worsen the progression of the HIV infection and increases HCV transmission. Therefore, clinical management and treatment of HCV is a priority in medical facilities that receive HIV infected patients. Clinical management of these patients involves specific diagnostic procedures and appropriately trained medical staff. The indication of treatment should meet specific clinical and laboratory criteria. There are a number of drugs currently available to treat hepatitis C in co-infected patients. PMID- 15880216 TI - Molecular and seroepidemiologic studies of Enterovirus 71 infection in the State of Para, Brazil. AB - In many countries, the Enterovirus 71 (EV-71) Picornaviridae family is associated to hand, foot and mouth disease in addition to acute neurological diseases while in Brazil these viruses are more closely associated to the latter group. The aim of this research was to use the first EV-71 isolate of the Northern region of Brazil in molecular and seroepidemiologic studies. Two (2.2%) out of 88 stool samples (44 cases of AFP), collected from January 1998 to December 2000 were positive for EV-71 isolation (73442/PA/99). Nucleotide sequence of the gen that codifies the VP1 protein showed that isolate 73442/PA/99 was similar to the EV-71 strains belonging to genotype B - more closely identified with EV-71 from North America. Neutralization test with 389 sera samples collected from January 1998 to November 2001, from individuals ranging from 0 to 15 years of age living in the city of Belem, State of Para showed the following results in relation to isolate 73442/PA/99 and prototype BrCr: a total of 207 individuals (53.2%) had neutralization antibodies to both viruses, 167 (42.9%) had no antibodies and 15 showed the presence of neutralizing antibodies to one of the two viruses. Only 20.2% of the children aged 0 to 3 had neutralizing antibodies to EV-71, indicating that these children were more susceptible to the infection. Both the seroprevalence study and VP1 sequencing were important to demonstrate the spread and the molecular pattern of the EV-71 circulating in the Northern Region of Brazil. PMID- 15880217 TI - Rat hepatocyte invasion by Listeria monocytogenes and analysis of TNF-alpha role in apoptosis. AB - Listeria monocytogenes, etiological agent of severe human foodborne infection, uses sophisticated mechanisms of entry into host cytoplasm and manipulation of the cellular cytoskeleton, resulting in cell death. The host cells and bacteria interaction may result in cytokine production as Tumor Necrosis Factor (TNF) alpha. Hepatocytes have potential to produce pro-inflammatory cytokines as TNF alpha when invaded by bacteria. In the present work we showed the behavior of hepatocytes invaded by L. monocytogenes by microscopic analysis, determination of TNF-alpha production by bioassay and analysis of the apoptosis through TUNEL technique. The presence of bacterium, in ratios that ranged from 5 to 50,000 bacteria per cell, induced the rupture of cellular monolayers. We observed the presence of internalized bacteria in the first hour of incubation by electronic microscopy. The levels of TNF-alpha increased from first hour of incubation to sixth hour, ranging from 0 to 3749 pg/mL. After seven and eight hours of incubation non-significant TNF-alpha levels decrease occurred, indicating possible saturation of cellular receptors. Thus, the quantity of TNF-alpha produced by hepatocytes was dependent of the incubation time, as well as of the proportion between bacteria and cells. The apoptosis rate increased in direct form with the incubation time (1 h to 8 + 24 h), ranging from 0 to 43%, as well as with the bacteria : cells ratio. These results show the ability of hepatocyte invasion by non-hemolytic L. monocytogenes, and the main consequences of this phenomenon were the release of TNF-alpha by hepatocytes and the induction of apoptosis. We speculate that hepatocytes use apoptosis induced by TNF-alpha for release bacteria to extracellular medium. This phenomenon may facilitate the bacteria destruction by the immune system. PMID- 15880218 TI - Retrospective study of malaria prevalence and Anopheles genus in the area of influence of the Binational Itaipu Reservoir. AB - The importance of hydroelectric dams beside the human interchange in the maintenance of malarious foci and the occurrence of the Anopheles genus on the Binational Itaipu Reservoir were the main points of this retrospective study. Data were collected from existing registrations at National, State and Municipal Health Departments and literature systematic overview, from January 1984 to December 2003. The occurrence of some outbreak of malaria, mainly by Plasmodium vivax, and the prevalence of species of the Anopheles genus different from Anopheles darlingi in the region are discussed. The malaria in the left bank of Parana River is a focal problem, which must be approached locally through health, educational and social actions to prevent the continuity of outbreaks in the area. Concomitantly, it is necessary to plan and apply effective surveillance measures in the influence area of the Itaipu Reservoir. PMID- 15880219 TI - Melanoides tuberculatus (Gastropoda: Thiaridae) as intermediate host of Heterophyidae (Trematoda: Digenea) in Rio de Janeiro metropolitan area, Brazil. AB - In the late 1960s, Melanoides tuberculatus snails were introduced in Brazil from North/East Africa and Southeast Asia. The first records of specimens infected with cercariae were registered in Rio de Janeiro State in 2001. The present study reports the occurrence of M. tuberculatus infected with larval trematodes in Rio de Janeiro City. Bottom sediment was collected with dip nets and sieved through 0.25 inch-mesh screening. Snails were transported to the laboratory in vials with stream water, then measured and individually isolated in glass vials with distilled water. They were exposed to artificial light and temperature to induce cercarial emergence. The most actively emerging cercariae were processed by differential staining and silver nitrate impregnation methods. Negative snails were subsequently dissected. Approximately 700 snails were collected. Snail total lengths ranged from 1.2 to 3.3 cm. The prevalence rate was 15.76% although 53.76% of the snails were found infected in one of the sites. Infected snails were infected with rediae and pleurolophocercous cercariae. Cercarial morphology and chaetotaxy were consistent with those of the family Heterophyidae mostly due to the presence of median dorsal and ventral fins on the tail and the absence of CI dorsal sensory receptors. PMID- 15880220 TI - Effect of Cysticercus cellulosae fractions on the respiratory burst of pig neutrophils. AB - Neutrophils, eosinophils and macrophages are cells that interact with invading parasites and naive hosts have been shown to have anti-parasitic activity. The initial reaction of these leukocytes is the generation of reactive oxygen species (ROS) to play in parasite expulsion. The present work was carried out to study the effect of total extract, scolex and membrane fractions from Cysticercus cellulosae on respiratory burst by pig neutrophils. Hydrogen peroxide (H2O2) production by neutrophils incubated with metacestode fractions from C. cellulosae showed an increase of: 190% (total extract), 120% (scolex) and 44% (membrane). High antioxidant catalatic activity (33%, 28%, 28% by total extract, scolex and membrane, respectively) was observed in neutrophils incubated with metacestode fractions, which could be an attempt at self-protection. Scolex and membrane fractions increased the phagocytic capacity of neutrophils (44% and 28%, respectively). On the other hand, total cysticerci did not alter the phagocytosis, possibly due to modifications in membrane function, caused by high ROS production from neutrophils in the presence of total cysticerci. Total fraction from C. cellulosae is toxic for neutrophils as shown by the decrease in phagocytic capacity, probably caused by high levels of ROS formation. The difference in toxicity of total extract, scolex and membrane fractions on neutrophils can be explained by the presence of an antigenic effect of the vesicular fluid in the total extract of C. cellulosae. PMID- 15880221 TI - Oral cysticercosis: case report and review of the literature. AB - Cysticercosis is a condition that occurs when man is infested by the larvae of Taenia solium, acting as an intermediate host instead of definitive. Oral cysticercosis is a rare event, and it represents a difficulty in clinical diagnosis. A case of oral cysticercosis in a 23-year-old white female who presented a painless swelling in the dorsal portion of the tongue is reported. An excisional biopsy was performed and histopathological examination revealed a cystic cavity containing the tapeworm. PMID- 15880222 TI - Detection of EBV-DNA in serum samples of an immunosuppressed child during a three years follow-up: association of clinical and PCR data with active infection. AB - Twenty-four whole blood and serum samples were drawn from an eight year-old heart transplant child during a 36 months follow-up. EBV serology was positive for VCA IgM and IgG, and negative for EBNA-IgG at the age of five years old when the child presented with signs and symptoms suggestive of acute infectious mononucleosis. After 14 months, serological parameters were: positive VCA-IgG, EBNA-IgG and negative VCA-IgM. This serological pattern has been maintained since then even during episodes suggestive of EBV reactivation. PCR amplified a specific DNA fragment from the EBV gp220 (detection limit of 100 viral copies). All twenty-four whole blood samples yielded positive results by PCR, while 12 out of 24 serum samples were positive. We aimed at analyzing whether detection of EBV DNA in serum samples by PCR was associated with overt disease as stated by the need of antiviral treatment and hospitalization. Statistical analysis showed agreement between the two parameters evidenced by the Kappa test (value 0.750; p < 0.001). We concluded that detection of EBV-DNA in serum samples of immunosuppressed patients might be used as a laboratory marker of active EBV disease when a Real-Time PCR or another quantitative method is not available. PMID- 15880223 TI - Mediastinal histoplasmosis: report of the first two Brazilian cases of mediastinal granuloma. AB - This report documents the first two Brazilian cases of mediastinal granuloma due to histoplasmosis, presenting selected aspects on the diagnosis. Tissue samples revealing histoplasmosis were obtained from each of the patients by mediastinoscopy and thoracotomy. In the second patient, a subcarinal calcified mass eroded into the bronchial tree, leading to secondary bilateral aspiration pneumonitis one week after thoracotomy. Although rare, histoplasmosis should be included in the differential diagnosis of mediastinal granuloma, specially if there are calcifications greater than 10 mm in dimension. PMID- 15880224 TI - First record of Desmodus rotundus in urban area from the city of Olinda, Pernambuco, Northeastern Brazil: a case report. AB - The objective of this report is to describe the first record of Desmodus rotundus in urban area from the city of Olinda, Pernambuco State, Northeastern Brazil, and to draw attention to the possible risk of rabies transmission in this place. After the complaint of a dog owner who observed three bats attacking his dog, images registering attacks of D. rotundus were captured with a video camera. From 09:00 p.m. on 13 February 2004 to 04:00 a.m. of the next day, a high frequency of haematophagic activity and the presence of several bites on the dog's body were observed. This finding represents a serious risk to public health. Thus, it is necessary to further study the bat fauna, with special attention to their feeding behaviour in this place, in order to better know their biology and to adopt pertinent control measures. This is, to our knowledge, the first record of D. rotundus in urban area of Olinda. PMID- 15880225 TI - Carbohydrate assimilation profiles of Brazilian Candida dubliniensis isolates based on ID 32C system. AB - The purpose of the present study was to evaluate the identification of 19 Brazilian C. dubliniensis based on the biochemical profile exhibited when tested by the commercial identification kit ID 32C (bioMerieux). Thirteen of the isolates were rigorously identified as C. dubliniensis and the remaining isolates (six) were considered as having a doubtful profile but the software also suggested that there was 83.6% of chances for them to be C. dubliniensis. As well as pointed by the literature the identification obtained by phenotypic tests should be considered presumptive for C. dubliniensis due to variability of this new species. PMID- 15880226 TI - Serotyping and evaluation of the virulence in mice of Streptococcus suis strains isolated from diseased pigs. AB - A total of 110 strains of Streptococcus suis, isolated from diseased pigs in Brazil were serotyped and analyzed for virulence. Serotyping of the strains resulted in the following classification: 42 strains of serotype 2 (38.2%), 10 strains of serotype 14 (9.1%), seven strains of serotype 9 (6.4%), three strains each of serotype 7 and 11 (2.7%), two strains each of serotype 1 and 8 (1.8%) and one strain each of serotypes 1/2, 3, 5, 6 and 10 (0.9%). Cross reactions among serotypes 1, 14 and 7 were observed in 21 strains (19.1%). Only 41.9% of the strains were lethal for mice using the pathogenicity test. PMID- 15880227 TI - Screening for pulmonary tuberculosis in Teresopolis, RJ, Brazil: the search for respiratory symptomatic patients in emergency service of "Hospital das Clinicas de Teresopolis Costantino Ottaviano, Fundacao Educacional Serra dos Orgaos". AB - The aim of the present study was to investigate the detection percentage of tuberculosis among patients that are respiratory symptomatic (TB suspects). In this work, we present the preliminary results of research carried out at "Hospital das Clinicas de Teresopolis Costantino Ottaviano da Fundacao Educacional Serra dos Orgaos (FESO)" from November 2003 to April 2004. Among the 40 respiratory symptomatic individuals identified and referred to the Tuberculosis Control Program in Teresopolis , two (5.0%) were characterized as smear-positive. These results confirm reports in the literature and underscore the need for and importance of this strategy. PMID- 15880228 TI - Multidrug resistant Salmonella typhi and antimicrobial agent quality and bioavailability. PMID- 15880229 TI - Dengue: a review of the laboratory tests a clinician must know to achieve a correct diagnosis. AB - Dengue is the most important disease caused by an arbovirus (1, 2, 3 and 4 serotypes) worldwide, especially in the tropical and sub-tropical regions. Its clinical manifestations range from asymptomatic infections to a severe disease characterized by hemorrhage and shock. The incidence of dengue virus activity in the Americas has substantially increased from 1980 to 1994. In Brazil, the increase in the incidence of dengue is especially linked to the dissemination of Aedes aegypti. Thus, a rapid and accurate dengue diagnosis is of paramount importance for effective control of dengue outbreaks [8]. Five serological tests have been used for the diagnosis of dengue infection: hemagglutination-inhibition (HI), complement fixation (CF), neutralization test (NT), immunoglobulin M (IgM) capture enzyme linked immunosorbent assay (MAC-ELISA) and indirect immunoglobulin G ELISA. The limitations of these techniques are the high cross-reactivity observed with these tests. Four methods of viral isolation have been routinely used for dengue viruses: intracerebral inoculation of newborn mice, inoculation on mammalian cell cultures, intrathoracic inoculation of adult mosquitoes, and inoculation on mosquito cell cultures. In recent years, several new diagnostic techniques have been developed and have proven very useful in dengue diagnosis, such as: nucleic and acid hybridization, RT-PCR. Currently, dengue diagnosis is based on serology, viral isolation and RNA detection. Enzyme-linked immunosorbent assays (ELISA) are still the most widely used technique for serological diagnosis, but they do not identify the dengue virus serotype responsible for the current infection, so molecular techniques may soon assume a very important role in dengue diagnosis. RT-PCR is definitely the most satisfactory test that can be used on these infections, since it has been shown to be able to detect dengue viruses up to the 10th day after the onset of the symptoms. PMID- 15880230 TI - Establishment of the serologic testing algorithm for recent human immunodeficiency virus (HIV) seroconversion (STARHS) strategy in the city of Sao Paulo, Brazil. AB - Several strategies aim at characterizing the AIDS epidemic in different parts of the world. Among these, the identification of recent HIV-1 infections using the recently described serologic testing algorithm for recent human immunodeficiency virus (HIV) seroconversion (STARHS) strategy was employed in four testing sites of the City of Sao Paulo Public Health Department (CSPPHD). Those identified as recently infected were invited to participate in a prospective clinical and laboratory evaluation study. We describe the establishment of the patient identification network and the success in enrolling the participants, as well as their clinical and laboratory characteristics. From May to December 2002, 6,443 persons were tested for HIV in the four participating sites, of whom 384 (5.96%) tested HIV-1 positive; 43 (11.2%) of them were identified as recently infected. Twenty-two were successfully enrolled in the follow-up study, but three of them did not meet clinical and/or laboratory criteria for recent HIV-1 infection. After these exclusions, the laboratory findings revealed a median CD4+ T lymphocyte count of 585 cells/microL (inter-quartile range 25-75% [IQR], 372 754), a CD8+ T lymphocyte count of 886 cells/microL (IQR, 553-1098), a viral load of 11,000 HIV-RNA copies/mL (IQR, 3,650-78,150), log10 of 4.04 (IQR 3.56-4.88). The identification of recent HIV infections is an extremely valuable way to evaluate the spread of the virus in a given population, especially when cohort studies, considered the gold standard method to evaluate incidence, are not available. This work demonstrated that establishing a network to identify such patients is a feasible task, even considering the difficulties in a large, resource-limited country or city. PMID- 15880231 TI - Production of monoclonal antibody to subtype 9 of Neisseria meningitidis and the distribution of this subtype in Brazil. AB - A new monoclonal antibody (5F81A4P1.9), which is specific for subtype 9 antigen of meningococci, was studied. The antibodies were raised against a previously non typable (NT) serogroup B strain from Brazilian patients and were found to react with the subtype antigen of prototype reference strains for subtype 9 (M982), as well as with those of homologous strains. The subtype 9 epitope was found in 6.8% of serogroup B strains among 602 strains of Neisseria meningitidis case isolates, including representative isolates from Brazilian states. Subtype P1.9 was predominantly related to serogroup B in Brazil among the isolates collected during the N. meningitidis epidemic in 1992. No significant differences were observed in the occurrence of subtype P1.9 among strains isolated from several Brazilian states. Fluorescence-activated cell-sorter analysis showed that 5F81A4 MAb recognized a 46 kDa protein on the surface of a homologous strain of N. meningitidis (B:4:P1.9). These results, in association with a bactericidal activity assay for 5F81A4, and with experimental passive protection in mice, demonstrated the importance of subtype 9 class 1 proteins of N meningitidis in Brazil. Serotyping is essential for the development of vaccination strategies. PMID- 15880232 TI - Improving survival among Brazilian children with perinatally-acquired AIDS. AB - Brazil was the first developing country to provide free, universal access to antiretroviral treatment for AIDS patients. The Brazilian experience thus provides the first evidence regarding the impact of such treatment on the survival of perinatally acquired AIDS cases in the developing world. MATERIAL AND METHODS: This retrospective cohort study used medical record reviews to examine characteristics and trends in the survival of a representative sample of 914 perinatally acquired AIDS cases in 10 Brazilian cities diagnosed between 1983 and 1998. RESULTS: Survival time increased steadily and substantially. Whereas half of the children died within 20 months of diagnosis at the beginning of the epidemic, 75% of children diagnosed in 1997 and 1998 were still alive after four years of follow-up. CONCLUSIONS: Advances in management and treatment have made a great difference in the survival of Brazilian children with AIDS. These results argue strongly for making such treatment available to children in the entire developing world. PMID- 15880233 TI - Risk of tuberculosis among household contacts in Salvador, Bahia. AB - Tuberculosis is one of the most important infectious diseases in the world. Only 68% of the estimated new tuberculosis (TB) cases in Brazil are diagnosed. Our aim was to determine the risk of infection among household contacts. STUDY DESIGN: Cohort of tuberculin-negative household contacts followed for 12 months. METHODS: Household contacts of randomly selected index acid-fast bacilli (AFB)-positive TB cases were evaluated through clinical examination, thorax X-ray, tuberculin, AFB smear and culture. Contacts with a negative response to the tuberculin test (less than 10 mm diameter) were retested after 90 days. Tuberculin reversal (used as a parameter of infection risk) was defined as an increase of at least 10 mm from the last measurement. RESULTS: 269 household contacts were followed. The prevalence of disease in this population was 3.7%. The prevalence of infection after the 12-month follow-up period was 63.9%. The risk of infection was 31.1% within 120 +/- 48 days. CONCLUSION: Household contacts of AFB positive tuberculosis patients have a very high prevalence and risk of tuberculosis infection. TB preventive or therapeutic measures directed towards this group should be implemented in Brazil. PMID- 15880234 TI - The influence of occult infection with hepatitis B virus on liver histology and response to interferon treatment in chronic hepatitis C patients. AB - Occult hepatitis B virus (HBV) infections have been identified in patients with chronic hepatitis C virus (HCV) infection, although the clinical relevance of occult HBV infection remains controversial. We searched for serum HBV DNA in 106 HBsAg negative/anti-HBc positive patients with chronic HCV infection and in 150 blood donors HBsAg negative/anti-HBc positive/anti-HCV negative (control group) by nested-PCR. HCV genotyping was done in 98 patients and percutaneous needle liver biopsies were performed in 59 patients. Fifty-two patients were treated for HCV infection with interferon alone (n=4) or combined with ribavirin (n=48) during one year. At the end and 24 weeks after stopping therapy, they were tested for HCV-RNA to evaluate the sustained virological response (SVR). Among the 106 HCV-positive patients, 15 (14%) were HBV-DNA positive and among the 150 HCV negative blood donors, 6 (4%) were HBV-DNA positive. Liver biopsy gave a diagnosis of liver cirrhosis in 2/10 (20%) of the HBV-DNA positive patients and in 6/49 (12%) of the HBV-DNA negative patients. The degree of liver fibrosis and portal inflammation was similar in HCV-infected patients HBV-DNA, irrespective of HBV-DNA status. SVR was obtained in 37.5% of the HBV-DNA positive patients and in 20.5% of the HBV-DNA negative patients; this difference was not significant. In conclusion, these data suggested that occult HBV infection, which occurs at a relatively high frequency among Brazilian HCV-infected patients, was not associated with more severe grades of inflammation, liver fibrosis or cirrhosis development and did not affect the SVR rates when the patients were treated with interferon or with interferon plus ribavirin. PMID- 15880235 TI - Identification of Leishmania (Leishmania) chagasi isolated from healthy skin of symptomatic and asymptomatic dogs seropositive for leishmaniasis in the municipality of Rio de Janeiro, Brazil. AB - Euthanasia of seropositive dogs has been one of the principal measures adopted by the Program for the Control of Leishmaniasis in Brazil for many years. However, its efficacy is currently being questioned. We obtained intact skin samples from 20 Leishmania-reactive dogs from the municipality of Rio de Janeiro that had been referred for euthanasia. The promastigote forms of Leishmania were isolated in culture from 18 of these animals. Fourteen of these isolates were identified as Leishmania (Leishmania) chagasi by isoenzyme electrophoresis; seven of these were from asymptomatic dogs and seven were from symptomatic animals with visceral leishmaniasis. In conclusion, cutaneous parasitism is found in the intact skin of dogs naturally infected with L. (L.) chagasi, irrespective of the presence or absence of clinical signs suggestive of visceral leishmaniasis. PMID- 15880236 TI - Evaluation of central supply units in public dental medicine colleges in Brazil. AB - A guarantee of quality of all steps involved in processing dental materials is essential to achieve security. A descriptive study was made of how Public Dental Medicine Colleges in Brazil process critical materials and determine the patterns of physical, chemical and biological control in their use of hot air ovens and autoclaves to sterilize these materials. The data were obtained with a questionnaire, sent by mail and analyzed with the software EPI-INFO 6.04. Among the 40 Brazilian public Dental Medicine Colleges, only 16 returned the questionnaire. In eight of these, the individuals responsible for the materials and sterilization center had a college degree in a human health field. In 14 institutions, the students were responsible for the cleaning of the instruments, but in six of these they did so outside of the materials and sterilization center. Both the autoclave and the dry heat oven were the method of choice in 13 of the 16 schools. The sterilization routine was routinely monitored by 11 of the institutions. Chemical control through the tape test in the autoclave was used by 13 of the schools, three institutions reported preventive maintenance, and biological indicators were used by seven of the 16 schools. Autoclaves are widely used because of the degree of biological security that this method offers, however physical, chemical and biological controls have not been routinely implemented by most of the institutions. PMID- 15880237 TI - Emerging acute Chagas disease in Amazonian Brazil: case reports with serious cardiac involvement. AB - Four cases of serious cardiac attacks by autochthonous Trypanosoma cruzi infection from the Brazilian Amazon are reported; three of them occurred in micro epidemic episodes. The manifestations included sudden fever, myalgia, dyspnea and signs of heart failure. Diagnosis was confirmed by specific exams, especially QBC (Quantitative Buffy Coat) and natural xenodiagnosis. Despite treatment with benznidazol, three patients died with serious myocarditis, renal failure and cardiac tamponade. The authors call attention to the emergence of this disease and reveal a previously unknown pathogenicity of T. cruzi strains in this area, added to a non-usual transmission form. PMID- 15880238 TI - Dengue hemorrhagic fever and acute hepatitis: a case report. AB - Dengue fever is the world's most important viral hemorrhagic fever disease, the most geographically wide-spread of the arthropod-born viruses, and it causes a wide clinical spectrum of disease. We report a case of dengue hemorrhagic fever complicated by acute hepatitis. The initial picture of classical dengue fever was followed by painful liver enlargement, vomiting, hematemesis, epistaxis and diarrhea. Severe liver injury was detected by laboratory investigation, according to a syndromic surveillance protocol, expressed in a self-limiting pattern and the patient had a complete recovery. The serological tests for hepatitis and yellow fever viruses were negative. MAC-ELISA for dengue was positive. PMID- 15880239 TI - An unusual case of ectopic tungiasis with pseudoepitheliomatous hyperplasia. AB - Tungiasis is caused by the penetration of the female sand flea Tunga penetrans into the epidermis, and subsequent hypertrophy of the parasite. In most cases lesions are confined to the feet. During a cross-sectional study, an unusual case of ectopic tungiasis in the inguinal area was detected. Histological examination of tissue samples showed a remarkable pseudoepitheliomatous aspect of the epidermis. Clinical features and differential diagnoses are discussed. PMID- 15880240 TI - Tribute to a master. PMID- 15880242 TI - Partnership between universities and the local healthcare system to benefit the town of Buriticupu, a poor community in the state of Maranhao, Brazil. PMID- 15880243 TI - Oxytalan elastic and collagen fibers during the repair process in experimental nitric oxide inhibition. AB - PURPOSE: To evaluate the repair process in rats with experimentally induced arterial hypertension. This study aimed to evaluate lesions in the ventricular myocardium and the repair process during experimental hypertension induced by systemic blockage of nitric oxide using N-omega-nitro-L-arginine methyl ester hydrochloride (L-NAME). Nitric oxide is an endothelial vasorelaxing factor and is necessary for the maintenance of normal arterial pressure, and L-NAME is an analog and antagonist of L-arginine, the substrate of the nitric oxide synthase. MATERIALS AND METHODS: We used 26 normotensive young male Wistar rats belonging to several litters. Animals were treated with oral administration of L-NAME dissolved in water (75 mg/100 mL) for 43 days. Hearts were weighed and processed by routine methods. Special stains utilized were Gomori's trichrome (aniline blue), picrosirius red polarization to identify fibrillar collagen, alcian blue technique (pH 0.5 and pH 2.5) to identify glycosaminoglycans, periodic acid Schiff technique (with and without amylases) to identify proteoglycans, and Weigert's resorcinol fuchsin solution (with and without oxone) to identify elastic fibers. RESULTS: The results showed significant elevation of the arterial pressure (P <0.01) and significant increase of cardiac weight (P <.0001) in the L NAME (hypertensive) treated group, as compared to an untreated control group. The histological analysis demonstrated wide infarcted myocardial areas in animals with nitric oxide blockade; several vascular changes such as thickening of the muscular tunica with fibrosis; thickening in the wall of small arteries and arterioles; and fibrinoid necrosis in the wall to nearly complete luminal obliteration. Reparative fibrosis involved mainly oxytalan elastic and collagen fibers. CONCLUSION: Oxytalan elastic and collagen fibers are of great importance for the postinfarct repair process occurring during experimental nitric oxide inhibition. PMID- 15880244 TI - Analysis of distortions in children with and without phonological disorders. AB - PURPOSE: To verify using 4 different tests the incidence of distortions in children with and without phonological disorders. METHOD: Forty children between 4 and 10.2 years of age, divided into 2 groups: 20 with normal development and 20 with phonological disorders. All children underwent the phonology tests of the Child Language Assessment ABFW and 2 spontaneous speaking tests to assess for phonologic alterations. After recording, the data were printed, analyzed, and classified according to the distortions. Nonparametric (Mann-Whitney) statistical analysis was performed with the significance level being set at P <.05. RESULTS: The phonological disorder group had significantly more occurrences of distortions in all tests compared to the control group (naming, P = .04; imitation P <.001; spontaneous speaking 1, P = .01; and spontaneous speaking 2, P = .002. The Pearson correlation coefficients of the distortion occurrences among the 4 tests were high. CONCLUSION: The phonological disorder group presented a greater number of distortions in all tests. The most frequent ones were /s, z, 3/ and the variability found within the phonological disorder group was very high. The children in this group had unstable phonological systems and so presented a high number of different distortions. Regarding the evaluation of the phonologic system, all the tests were good evaluation methods since the correlations between them were high. PMID- 15880245 TI - Origin of adenocarcinoma in Barrett's esophagus: p53 and Ki67 expression and histopathologic background. AB - Barrett's esophagus is the substitution of squamous epithelium of the distal esophagus by columnar epithelium. Intestinal metaplasia in Barrett's esophagus is considered to be the main risk factor for the development of adenocarcinoma. Diffuse adenocarcinoma and Barrett's esophagus without intestinal metaplasia are rare, and reports on the subject are scarce. PURPOSE AND METHOD: To estimate the prevalence of adenocarcinoma in 297 patients with Barrett's esophagus, during the period of 1990 to 2002, and in 13 patients undergoing surgery, to conduct detailed macroscopic and microscopic analysis, with performance of immunohistochemical tests for p53 and Ki67, correlating the type of tumor with its adjacent epithelium. RESULTS: In our patients with Barrett's esophagus, there was a prevalence of 5.7% of adenocarcinoma. The tumors developed only when the Barrett's esophagus segment was long (>3.0 cm). Tumors were located close to the squamous-columnar junction. The histological study revealed 2 patients (15.4%) with Barrett's esophagus adjacent to a tumor with gastric metaplasia without the presence of intestinal metaplasia. Tumors were classified according to Nakamura's classification (23% differentiated pattern, and 77% undifferentiated pattern) and to Lauren's classification (61% intestinal and 39% diffuse). The difference is due to the migration of microtubular and foveolar tumors of undifferentiated (gastric) pattern in Nakamuras classification to the Lauren's intestinal type. The immunohistochemical test for Ki67 was strongly positive in all the patients, thus evidencing intense cell proliferation in both the columnar epithelium and tumor. Expression of p53 was negative in 67% of the adjacent columnar epithelia and 42% of the tumors, without any correlation between the tissue types. CONCLUSION: Adenocarcinoma develops from mixed columnar epithelium, either intestinal or gastric, showing both the gastric and the intestinal patterns; thus, tumors can also grow in columnar epithelium without intestinal metaplasia. Barrett's esophagus should be followed up for the possibility of progression to malignancy, especially when the segment is longer than 3 cm. PMID- 15880246 TI - Effects of corticosteroids in very low birth weight newborns dependent on mechanical ventilation. AB - Corticosteroids have been used in bronchopulmonary dysplasia prevention because of their antiinflammatory effects. Among their effects is a decrease in the incidence of bronchopulmonary dysplasia. However, short- and long-term side effects have been detected in preterm newborns. PURPOSE: To analyze the effects of corticosteroids on bronchopulmonary dysplasia, length of stay, mortality, growth, as well as the adverse effects in very low birth weight newborns between 10 and 14 days of life and dependent on mechanical ventilation. METHODS: Cohort study. All newborns with a birth weight under 1500 g, mechanical ventilation dependent between 10 and 14 days of life, during the period January 2000 and June 2001 were included (n = 38). They were divided into 2 groups: Group I with corticosteroids (n = 16) and Group II without corticosteroids (n = 22). Dexamethasone administration: from the 10th day of life, d1-d3, 0.3 mg/kg/d; d4 d6, 0.2 mg/kg/d; d7-d9, 0.1 mg/kg/d. Respiratory evolution, bronchopulmonary dysplasia (oxygen dependence at 28 days of life), growth pattern and the presence of adverse effects were analyzed. RESULTS: The incidence of bronchopulmonary dysplasia was 6.5% (Group I) and 30% (Group II), P = .07. A decrease in growth was detected in Group I compared with Group II (change in weight: Group I--47 g/week, Group II--85.5 g/week, P = .06; change in head circumference: Group I- 0.75 cm/week, Group II--1 cm/week, P = .05). CONCLUSION: Use of corticosteroids in very low birth weight infants dependent on mechanical ventilation during the first 10 to 14 days of life did not affect the respiratory evolution and occurrence of bronchopulmonary dysplasia, but the velocity of growth was reduced. PMID- 15880247 TI - Relationship between surgical procedure and outcome for patients with grade I chondrosarcomas. AB - PURPOSE: To evaluate the oncological outcome of patients with grade I chondrosarcomas according to the type of surgical treatment performed, since there is still controversy regarding the need for aggressive resections to reach a successful outcome. MATERIALS AND METHODS: The records of 23 patients with grade I chondrosarcomas were reviewed. The mean age was 38.4 years, ranging from 11 to 70 years; 52% were men and 48% were women. The femur was the site of 13 tumors. The tumors were staged as IA (17, 74%) and IB (6, 26%). Regarding tumor location, 74% (17) were medullary, 22% (5) were peripheral, and 4% (1) was indeterminate. Tumor size ranged from 2 to 25 cm, mean 7.9 cm. Regarding the surgical procedure, 11 patients underwent intralesional resection, 9 patients underwent wide resection, and 3 underwent radical resection. The follow-up period ranged from 24 to 192 months. RESULTS: None of the patients developed local recurrence or metastases; 7 patients had other general complications. CONCLUSIONS: This data supports the use of less aggressive procedures for treatment of low-grade chondrosarcomas. PMID- 15880248 TI - Complement levels in Brazilian children during and after meningococcal meningitis. AB - PURPOSE: To evaluate the functional activity of the classical and alternative pathways of the complement system and the levels of C3, C4, and factor B during the first episode of meningococcal infection and during the convalescence period. PATIENTS AND METHODS: Ten Brazilian children ranging in age from 8 months to 8 years, admitted from 1991 to 1993 with a clinical-laboratory diagnosis of meningococcal meningitis, were studied during acute infection (up to 7 days from diagnosis) and during the convalescence period (1 to 6 months after the acute episode). C3, C4, and Factor B were measured using nephelometry, and the lytic activity of classical and alternative pathways were evaluated by a kinetic method and expressed as the time needed to lyse 50% of an erythrocyte suspension (T1/2, expressed in seconds). Low T1/2 values for classical and alternative pathways correlate with high activities of the classical and alternative complement pathways, respectively. RESULTS: A significant difference was observed between the alternative pathway lytic activity during infection and the convalescence period (282 vs 238 seconds, respectively, P = .01). No differences were detected in the other complement parameters analyzed. CONCLUSIONS: In the presence of meningococcal meningitis, the alternative pathway is preferentially activated. This is probably due to the greater ability of the meningococcal endotoxin to activate this pathway in vivo. PMID- 15880249 TI - Frequency of the deltaF508 mutation in 108 cystic fibrosis patients in Sao Paulo: comparison with reported Brazilian data. AB - PURPOSE: To analyze the frequency of the delta F508 (deltaF508) deletion mutation in 108 unrelated cystic fibrosis patients and compare the results with the previously reported data for Brazilian patients. Cystic fibrosis is the leading cause of genetic disease in Caucasians, and the deltaF508 deletion is the most common mutation associated with the disease. METHOD: The frequency of the deltaF508 mutation was assessed by means of a polymerase chain reaction (PCR) followed by detection in 8% silver-stained polyacrylamide gels. RESULTS: Twenty three of 108 patients (21.3%) were homozygous for the deltaF508 deletion, 50 were heterozygous (46.3%), and the remaining 35 (32.4%) were non-carriers. In terms of alleles, there were 96 mutated (96/216 or 44.45%) and 120 wild-type ones (120/216 or 55.5%). CONCLUSION: The 44.45% of affected alleles that were found is higher than the 33% first described in 1993, but slightly lower than the 48% recently reported. Moreover, our data corroborated the idea that the frequency of the deltaF508 mutation is lower in Brazil in comparison to that found in studies carried out in Europe and North American (circa 70.0%), probably due to increased racial miscegenation. These findings must be taken into account before any genetic screening of the population is proposed in Brazil. PMID- 15880250 TI - Quality of life and depression in patients undergoing total and partial laryngectomy. AB - PURPOSE: The surgical treatment of head and neck cancer, primarily laryngeal cancer, causes sequelae and can change the patient's quality of life. The purpose of this study was to investigate the impact of laryngectomy on the quality of life regarding the functional, physical, psychological, and social aspects. METHODS: Fourteen patients underwent total laryngectomy and 16 underwent vertical partial laryngectomy. The Quality of Life Core Questionnaire (QLQ-C30) and Head and Neck (H&N35) questionnaire from the European Organization for Research and Treatment of Cancer (EORTC) were used for quality of life evaluation, while the Beck Depression Inventory questionnaire was used for the depression screen. RESULTS: In the total laryngectomy group, reported adverse effects were worsened, social and emotional function (21.3%), olfaction and taste changes (85.6%), cough (71.3%), speech difficulty (100%), and dysphagia (64.3%). Most of the patients (85.5%) judged their quality of life to be reasonable. In the partial laryngectomy group, reported adverse effects were worsened, emotional function (71.4%), speech difficulty (100%), and dysphagia (31.3%). However, most of the patients judged their quality of life to be above the general average. CONCLUSION: Despite being different surgeries, both groups experienced similar difficulties but at different levels. The quality of life was judged worse in the patients who underwent total laryngectomy. PMID- 15880251 TI - What is behind a student's choice for becoming a doctor? AB - PURPOSE: To determine the reasons for choosing the medical profession by interviewing freshmen medical students from the Faculty of Medicine, University of Sao Paulo and investigating their socio-economic and psychological profiles, as well as to determine whether there are gender differences. METHOD: One hundred and sixty three freshmen medical students answered a questionnaire regarding their socio-economical profile. Of those, 30 female and 30 male students underwent a face-to-face interview regarding the career choice, the Thematic Apperception Test (TAT) and the 16 Personality Factor Questionnaire (16 PF). The results were analyzed qualitatively and by Mann-Whitney, chi2, Fisher's Exact, and r(p) coefficient tests. RESULTS: Most students were middle class, catholic, and had physicians in their families. Students of both genders had made an: early choice of the medical career and a persistence in taking exams many times, even after being discouraged. They also showed an awareness of the difficulties and limitations in developing their careers. The study revealed a strong valuation of the humanistic aspects of medicine; openness to new experiences; a deep personal identification with the choice of profession; a critical need for fulfillment in their careers; and conscious and unconscious desires to help people and be recognized for their usefulness, without being narcissistic. Female students were more sensitive (P <.001) and less imaginative (P = .005) than male students, who were more utilitarian and less grounded; female students tended to present greater emotional maturity while male students presented a greater tendency towards competition, and were more ambitious. CONCLUSIONS: Students of both genders have similar socio-economical profiles and features regarding their motivations for choosing the medical profession. Slight differences were found regarding some psychological aspects. PMID- 15880252 TI - Effects of an aquatic versus non-aquatic respiratory exercise program on the respiratory muscle strength in healthy aged persons. AB - Aging of the worldwide population is a concern of most governmental entities, spanning practically all areas of prevention and rehabilitation. Aging leads to physiological alterations that result in adverse social and financial effects. There is a trend to emphasize prevention, which is less expensive and socially more desirable than therapeutic intervention. PURPOSE: To assess the effect of a program of aquatic versus non-aquatic respiratory exercises on respiratory muscle strength in healthy aged persons. METHODS: The respiratory muscle strength was measured in 81 subjects between 60 and 65 years, 59 of which completed the program. Subjects were randomized into 3 groups. G(aquatic) undertook a program of respiratory exercise in an aquatic environment. G(non-aquatic) undertook the same program in a non-aquatic environment. G(control) acted as the negative control. Programs were applied three times a week for 10 consecutive weeks. Subsequently, subjects were reevaluated, and results compared to each individual's pre-treatment own result and between the groups. The data were statistically analyzed using the paired t test and the Sign test. Comparisons between the groups were performed through parametric and nonparametric variance. A comparison of G(aquatic) and G(non-aquatic) versus G(control) was performed using the Dunnett test. RESULTS: A significant improvement in the inspiratory muscle strength in the G(aquatic) group compared to the G(control), group was found, suggesting beneficial effects mediated by the aquatic exercise. The expiratory muscles did not show significant alterations. CONCLUSION: Aquatic respiratory exercise improves the inspiratory muscle strength of healthy aged persons. However, neither aquatic nor non-aquatic respiratory exercise influences the expiratory muscle strength. PMID- 15880253 TI - Small volume hypertonic resuscitation of circulatory shock. AB - Small volume hypertonic resuscitation is a relatively new conceptual approach to shock therapy. It was originally based on the idea that a relatively large blood volume expansion could be obtained by administering a relatively small volume of fluid, taking advantage of osmosis. It was soon realized that the physiological vasodilator property of hypertonicity was a useful byproduct of small volume resuscitation in that it induced reperfusion of previously ischemic territories, even though such an effect encroached upon the malefic effects of the ischemia reperfusion process. Subsequent research disclosed a number of previously unsuspected properties of hypertonic resuscitation, amongst them the correction of endothelial and red cell edema with significant consequences in terms of capillary blood flow. A whole set of actions of hypertonicity upon the immune system are being gradually uncovered, but the full implication of these observations with regard to the clinical scenario are still under study. Small volume resuscitation for shock is in current clinical use in some parts of the world, in spite of objections raised concerning its safety under conditions of uncontrolled bleeding. These objections stem mainly from experimental studies, but there are few signs that they may be of real clinical significance. This review attempts to cover the earlier and the more recent developments in this field. PMID- 15880254 TI - Increased arterial distensibility and renovascular hypertension in Goldenhar syndrome. AB - This is a report of the successful angioplastic treatment of an association of renovascular hypertension with renal artery stenosis and the Goldenhar syndrome (a variant of oculoauriculovertebral dysplasia). For the first time to date, this association, which occurred in a 13-year-old girl, is reported. Additionally, increased arterial distensibility in spite of arterial hypertension was detected by noninvasive methods. The similarity of this finding and in those for other genetic diseases, suggests that the vascular lesions could be linked to the Goldenhar syndrome. PMID- 15880255 TI - Cervical necrotizing fasciitis due to bacterial tonsillitis. AB - Necrotizing fasciitis is a severe and potentially fatal soft tissue infection, but involvement of the head and neck is rare. We report on 4 cases of cervical necrotizing fasciitis arising from tonsillitis. One patient was diabetic and one had received steroids before disease development. One patient developed acute respiratory failure and died of septic shock. Three patients recovered, helped by early recognition, aggressive surgical intervention, appropriate broad-spectrum antibiotics, and supportive therapy. The common bacteria found in all abscess samples were Streptococcus sp., but mixed flora with anaerobic organisms was seen in all but 1 case. Tonsillitis and peritonsillar abscess must be suspected as a cause of cervical necrotizing fasciitis and a successful result can be achieved with immediate aggressive treatment. PMID- 15880256 TI - Structural features and molecular evolution of Bowman-Birk protease inhibitors and their potential application. AB - The Bowman-Birk inhibitors (BBIs) are well-studied serine protease inhibitors that are abundant in dicotyledonous and monocotyledonous plants. BBIs from dicots usually have a molecular weight of 8k and are double-headed with two reactive sites, whereas those from monocots can be divided into two classes, one approximately 8 kDa in size with one reactive site (another reactive site was lost) and the other approximately 16 kDa in size with two reactive sites. The reactive site is located at unique exposed surfaces formed by a disulfide-linked beta-sheet loop that is highly conserved, rigid and mostly composed of nine residues. The structural features and molecular evolution of inhibitors are described, focusing on the conserved disulfide bridges. The sunflower trypsin inhibitor-1 (SFTI-1), with 14 amino acid residues, is a recently discovered bicyclic inhibitor, and is the most small and potent naturally occurring Bowman Birk inhibitor. Recently, BBIs have become a hot topic because of their potential applications. BBIs are now used for defense against pathogens and insects in transgenic plants, which has advantages over using toxic and polluting insecticides. BBIs could also be applied in the prevention of cancer, Dengue fever, and inflammatory and allergic disorders, because of their inhibitory activity with respect to the serine proteases that play a pivotal role in the development and pathogenesis of these diseases. The canonical nine-residue loop of BBIs/STFI-1 provides an ideal template for drug design of specific inhibitors to target their respective proteases. PMID- 15880257 TI - Searching for potential drug targets in two-component and phosphorelay signal transduction systems using three-dimensional cluster analysis. AB - Two-component and phosphorelay signal transduction systems are central components in the virulence and antimicrobial resistance responses of a number of bacterial and fungal pathogens; in some cases, these systems are essential for bacterial growth and viability. Herein, we analyze in detail the conserved surface residue clusters in the phosphotransferase domain of histidine kinases and the regulatory domain of response regulators by using complex structure-based three-dimensional cluster analysis. We also investigate the protein-protein interactions that these residue clusters participate in. The Spo0B-Spo0F complex structure was used as the reference structure, and the multiple aligned sequences of phosphotransferases and response regulators were paired correspondingly. The results show that a contiguous conserved residue cluster is formed around the active site, which crosses the interface of histidine kinases and response regulators. The conserved residue clusters of phosphotransferase and the regulatory domains are directly involved in the functional implementation of two component signal transduction systems and are good targets for the development of novel antimicrobial agents. PMID- 15880258 TI - Identification of a novel nucleus protein involved in the regulation of urokinase in 95D cells. AB - The urokinase-type plasminogen activator (uPA) plays an important role in cellular invasion. By using the downstream part of a 74 bp DNA region called the cooperation mediator (COM) of the uPA promoter as a bait sequence in the yeast one-hybrid screen, a gene called PBK1 was previously cloned from the cDNA library of the 95D lung cancer cell strain. In this study, the intracellular distribution of PBK1 was studied by using the transient transfection of pEGFP-C3-PBK1, and PBK1 was found to be localized in the nucleus. Co-transfection of pEGFP-C3-PBK1 and the deletion mutants of the pGL3-uPA promoter indicated that PBK1 can increase the uPA promoter activity by about 25% and this effect is uPA enhancer dependent. Western blotting and Enzyme-linked immunoadsordent assay further confirmed that PBK1 can upregulate the expression of uPA. Our results suggest that PBK1 is involved in the regulation of uPA expression, which might provide a new clue to further understanding the regulation mechanism of uPA expression. PMID- 15880259 TI - Establishment of a screening system for selection of siRNA target sites directed against hepatitis B virus surface gene. AB - To study the inhibitory effects of plasmid-derived small interfering RNA (siRNA) and synthetic siRNA on the expression of the hepatitis B virus surface (HBs) gene, three plasmid-derived siRNAs and one synthetic siRNA that complement the coding region of the HBs gene were prepared. The HBs expression plasmid pHBs-EGFP was also constructed. HeLa cells were co-transfected with pHBs-EGFP and the above siRNAs. The HBs mRNA quantities were measured by reverse-transcription PCR, and the level of HBs-EGFP fusion protein was quantified by fluorescent microscope. The concentrations of the hepatitis B virus surface antigen (HBsAg) derived from the culture supernatant of transfected HepG2.2.15 cells were measured by an enzyme-linked immunosorbent assay (ELISA) kit. The results showed that the three plasmid-derived siRNAs and the synthetic siRNA can effectively reduce the quantities of HBs mRNA and protein. The plasmid-derived siRNA psiRNA1 was found to be the most effective inhibitor of HBs expression. It can inhibit HBs-EGFP expression by 63.3% and suppress HBs mRNA by 75.6%. To further substantiate the above observations, psiRNA1 was transfected into HepG2.2.15 cells (an HBV secreting cell line). The transfections resulted in almost complete blockage of HBsAg production, whereas control vector-transfected cells secreted high levels of HBsAg 7 days post-transfection. In conclusion, our data suggests that RNA interference (RNAi) is an efficient approach for reducing the level of HBs transcripts and proteins and for suppressing HBsAg production. PMID- 15880260 TI - Cloning and overexpression of CYP6F1, a cytochrome P450 gene, from deltamethrin resistant Culex pipiens pallens. AB - CYP6F1 (GenBank/EMBL accession No. AY662654), a novel gene with a complete encoding sequence in the cytochrome P450 family 6, was cloned and sequenced from deltamethrin-resistant 4th instar larvae of Culex pipiens pallens. The cDNA sequence of CYP6F1 has an open reading frame of 1527 bp, which encodes a putative protein of 508 amino acid residues. The deduced amino acid sequence of CYP6F1 indicated that the encoded P450 has conserved domains of a putative membrane anchoring signal, putative reductase-binding sites, a typical heme-binding site, an ETLR motif and substrate recognition sites. Semi-quantitative RT-PCR analysis indicated that the CYP6F1 gene was expressed to a greater extent in the deltamethrin-resistant strain than in the susceptible strain of Cx. pipiens pallens. The expression levels of the CYP6F1 gene in the deltamethrin-resistant 1st, 2nd, 3rd, 4th instar larvae and adult female mosquitoes differed, with highest expression levels in the 4th instar larvae. In addition, the CYP6F1 gene was stably expressed in mosquito C6/36 cells, and the expected 61.2 kDa band was identified by Western blotting. The cells transfected with CYP6F1 had an increased resistance to deltamethrin as compared with control cells. These results indicate that CYP6F1 is expressed at higher levels in the deltamethrin resistant strain, and may confer some insecticide resistance in Cx. pipiens pallens. PMID- 15880261 TI - Molecular cloning and mRNA profile of insulin-like growth factor type 1 receptor in orange-spotted grouper, Epinephelus coioides. AB - The insulin-like growth factor type 1 receptor (IGF-IR) belongs to the tyrosine kinase (TK) receptor family. Besides being mitogenic, IGF-IR plays a crucial role in cell survival, transformation and maintenance of the malignant phenotype. In this study, we cloned the cDNA from the hypothalamus of the orange-spotted grouper (Epinephelus coioides) using reverse transcription PCR (RT-PCR) and the rapid amplification of cDNA ends (RACE) method. The deduced amino acid sequence showed that the receptor comprises 1413 amino acid residues. It contains cysteine rich domains in its alpha-subunit, and a conserved transmembrane domain and TK domains in its beta-subunit. Comparison of the amino acid sequence with those of other species showed that the grouper IGF-IR shares 90.2%, 89.6%, 71.9% and 72% similarity with the IGF-IR of the Japanese flounder, turbot, zebrafish-a and zebrafish-b, respectively. When compared with its mammalian homologue, grouper IGF-IR contains a large insertion at its C-terminus. Phylogenetic analysis has revealed that the grouper IGF-IR belongs to the b-type IGF-IRs and has a higher similarity with flounder and turbot IGF-IR, and a lower similarity (<70%) with human, mouse and avian IGF-IR. Grouper IGF-IR transcripts were detected in the brain, peripheral tissues, embryos and early development larvae by semi quantitative RT-PCR assay. It was observed that IGF-IR mRNA expression was greater in the brain than in peripheral tissues. The level of IGF-IR mRNA expression was much higher in retina, gonad, skeletal muscle and gill tissues than in liver, heart and thymus tissues. The expression of IGF-IR can be visualized as a ubiquitous signal in unfertilized eggs, embryos and early development larvae. The distribution pattern of IGF-IR mRNA in grouper development suggests that IGF-IR plays an important role in the embryo and early larval development stages. PMID- 15880262 TI - Up-regulation of NKX3.1 expression and inhibition of LNCaP cell proliferation induced by an inhibitory element decoy. AB - NKX3.1 is an androgen-regulated prostate-specific homeobox gene that is thought to play an important role in prostate development and cancerogenesis. NKX3.1 acts as a tumor suppressor gene specifically in the prostate. Up-regulation of NKX3.1 gene offers a promising gene therapy for prostate cancer. The decoy strategy has been developed and is considered a useful tool for regulating gene expression and gene therapy. In our previous studies, we identified a 20 bp inhibitory element upstream of the NKX3.1 promoter. In this study, we focused on using the 20 bp inhibitory element decoy to block negative regulation of the NKX3.1 gene and to up-regulate NKX3.1 expression using synthetic double-stranded oligodeoxynucleotides of the 20 bp inhibitory element. We found in an electrophoretic mobility shift assay experiment that the 20 bp inhibitory decoy presented competitive binding to a specific binding protein of the 20 bp inhibitory element in prostate cancer cell line LNCaP. In luciferase reporter gene assays, we found that the 20 bp inhibitory decoy could enhance NKX3.1 promoter activity, and RT-PCR and Western blot analysis revealed that NKX3.1 expression was up-regulated effectively by the transfection with the 20 bp inhibitory decoy. Furthermore, cell proliferation was inhibited by up-regulated NKX3.1 expression induced by the 20 bp inhibitory decoy. PMID- 15880263 TI - Comparative study of apoptosis-related gene loci in human, mouse and rat genomes. AB - Many genes are involved in mammalian cell apoptosis pathway. These apoptosis genes often contain characteristic functional domains, and can be classified into at least 15 functional groups, according to previous reports. Using an integrated bioinformatics platform for motif or domain search from three public mammalian proteomes (International Protein Index database for human, mouse, and rat), we systematically cataloged all of the proteins involved in mammalian apoptosis pathway. By localizing those proteins onto the genomes, we obtained a gene locus centric apoptosis gene catalog for human, mouse and rat. Further phylogenetic analysis showed that most of the apoptosis related gene loci are conserved among these three mammals. Interestingly, about one-third of apoptosis gene loci form gene clusters on mammal chromosomes, and exist in the three species, which indicated that mammalian apoptosis gene orders are also conserved. In addition, some tandem duplicated gene loci were revealed by comparing gene loci clusters in the three species. All data produced in this work were stored in a relational database and may be viewed at http://pcas.cbi.pku.edu.cn/database/apd.php. PMID- 15880264 TI - Prolonged Alzheimer-like tau hyperphosphorylation induced by simultaneous inhibition of phosphoinositol-3 kinase and protein kinase C in N2a cells. AB - Co-injection of wortmannin (inhibitor of phosphatidylinositol-3 kinase, PI3K) and GF109203X (inhibitor of protein kinase C, PKC) into the rat brain was found to induce spatial memory deficiency and enhance tau hyperphosphorylation in the hippocampus of rat brain. To establish a cell model with durative Alzheimer-like tau hyperphosphorylation in this study, we treated N2a neuroblastoma cells with wortmannin and GF109203X separately and simultaneously, and measured the glycogen synthase kinase 3 (GSK-3) activity by gamma-32P-labeling and the level of tau phosphorylation by Western blotting. It was found that the application of wortmannin alone only transitorily increased the activity of GSK-3 (about 1 h) and the level of tau hyperphosphorylation at Ser396/Ser404 and Ser199/Ser202 sites (no longer than 3 h); however, a prolonged and intense activation of GSK-3 (over 12 h) and enhanced tau hyperphosphorylation (about 24 h) were observed when these two selective kinase inhibitors were applied together. We conclude that the simultaneous inhibition of PI3K and PKC can induce GSK-3 overactivation, and further strengthen and prolong the Alzheimer-like tau hyperphosphorylation in N2a cells, suggesting the establishment of a cell model with early pathological events of Alzheimer's disease. PMID- 15880265 TI - Possible effect of 30K proteins in embryonic development of silkworm Bombyx mori. AB - The silkworm Bombyx mori possesses a 30K protein family of 3x10(4) Da, the biological functions of which have not been fully identified. The relationship between the 30K protein family and the embryonic development of temperature sensitive sex-linked mutant strain of silkworm was investigated by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The results show that protein spots 1-5 of the 30K protein family, mainly existing in normal strain, are possibly related to embryonic development. The early consumption of a 30K protein named 6G1-30K-1 and the accumulation of 30K proteins named 6G1-30K-3 and 6G1-30K-4 are likely caused by the destruction of physiological balance in normal embryonic development, which may lead to lower hatchability of the temperature sensitive strain. The results suggest that reasonable metabolism of 30K proteins is a prerequisite for the embryo's normal development. PMID- 15880266 TI - Method for the synthesis of phosphinic acids from hypophosphites. V. The synthesis of pseudo-alpha,alpha-dipeptides. AB - The method for the synthesis of 2-substituted 2-hydroxycarbonylethyl-1 aminoalkylphosphinic acids (I) (pseudo-alpha,alpha-dipeptides) from ammonium and potassium hypophosphites (II) is described. The proposed route to the synthesis of pseudo-alpha,alpha-dipeptides consists in addition hypophosphite to acrylic compounds and formation of the first phosphorus-carbon bond with following addition of aminoacid fragment and formation of the second phosphorus-carbon bond. The key intermediates of the synthesis - phosphonous acids (III) and their silylic esters (IV) were obtained at the first stage of the process as the result of the addition of the bis(trimethylsilyl)hypophosphite in situ to suitably substituted acrylates. The modificated procedure for the Kabachnik-Fields reaction of 2-substituted 2-alkoxycarbonylethyl phosphonous acids (III), acetamide, benzaldehyde in acetic anhydride with following hydrolysis results in 2-substituted 2-hydroxycarbonylethyl-alpha-aminobenzyl phosphinic acids (Ia-c) (pseudo-phenylglycylpeptides). Bis(trimethylsilyl) 2-substituted 2 alkoxycarbonylethylphosphonites (IV) in situ were added to N-tritylmethanimine and following alcoholysis and acid hydrolysis of addition products gave 2 substituted 2-hydroxycarbonylethyl-aminomethylphosphinic acids (Id-f) (pseudo glycylpeptides). PMID- 15880267 TI - Recent advances in asymmetric synthesis of pipecolic acid and derivatives. AB - This review covers the literature relating to asymmetric syntheses of pipecolic acid derivatives from 1997 to present. This review is organized according to the position and the degree of substitution of the piperidinic cycle. In a first section, syntheses of pipecolic acid itself are described. Then, successively, syntheses of C-3, C-4, C-5, C-6 substituted pipecolic acid derivatives are reported. Finally, syntheses of unsaturated pipecolic acid derivatives are presented before the last part devoted to the polysubstituted pipecolic acid derivatives. PMID- 15880268 TI - Problems in classifying vasculitis in children. AB - Vasculitis is inflammation of blood vessels, which may also affect the arteries of the kidney. The definition and classification of vasculitides derive from the Chapel Hill nomenclature criteria (CHCC) and American College of Rheumatology (ACR) criteria, respectively. These criteria are not perfect, and adult literature refers to problems with misclassification and a low concordance between the two criteria. The problems associated with defining a child patient according to these criteria are even more apparent, since both sets of criteria have been based on adult data alone and have never been validated in children. When classifying their patients as having polyarteritis, paediatricians do not keep strictly to the CHCC or ACR criteria. A recent series has shown that the microscopic polyarteritis associated with MPO-ANCA and the HBs associated with classic polyarteritis nodosa are rarer in children than in adults. Paediatricians should revise the existing criteria based on registries and international consensus in the field. PMID- 15880269 TI - The future of pediatric nephrology. AB - The delineation of renal disease in children dates to the 1880s with descriptions of Henoch's purpura, bladder exstrophy, renal rickets and nephritis. The discipline of pediatric nephrology mainly emerged during the 20th century in response to problems in fluid and electrolyte balance, characterization of the nephrotic syndrome, use of renal biopsy, antibiotic therapy of urinary tract infections, dialysis and transplantation in children, growth problems associated with chronic renal failure, detection and therapy of hypertension, and the creation of both national and international pediatric nephrology societies and a journal now in its 18th year. The development of molecular and cell biology, genetic and genomic techniques and bioinformatics methods underlie many future directions. We should anticipate further elucidation of single gene disorders, of complex trait analysis of disorders, such as diabetic nephropathy and hypertension, the interplay of developmental genes and gene products and interactions within the podocyte. Specific therapies directed against inflammation, vascular damage, cyst development, the ravages of proteinuria and graft rejection (or induction leading to tolerance) will be developed. Stem cell therapies may replace lost renal mass, even of specific nephron sites. Novel therapies will also modulate the cell cycle, tyrosine kinase signaling and apoptosis. In addition, drugs will be specifically tested in children for many renal conditions. Larger and more specialized registries will be developed; epidemiologic studies and exploration of large data sets will lead to clinical guidelines that are evidenced-based. There is a need for more careful measurement of glomerular filtration rate (GFR), proteinuria and cytokines, and a fuller appreciation of the nutritional and hormonal role of the kidney. Finally, the antecedents of adult renal disease and the need to intervene in a proactive fashion will be realized. Despite these impressive advances in care, the greatest challenges will be in providing children with renal disease access to well trained pediatric nephrologists, especially in Asia (1 billion children), Africa, Central and South America, and in immigrant and refugee populations. Included in this challenge is the capacity to have affordable access to use of contemporary techniques, and effective medications and prevention strategies. The International Pediatric Nephrology Association (IPNA), its journal, and pediatric advocates will need to use their energies to take on these challenges. PMID- 15880270 TI - Nontuberculous mycobacterial exit-site infection and abscess in a peritoneal dialysis patient. A case report and review of the literature. AB - Nontuberculous mycobacterial infections of peritoneal dialysis catheter exit sites have rarely been reported in patients on peritoneal dialysis. We report here a case of Mycobacterium abscessus exit site infection with abdominal wall abscess formation in an adolescent on peritoneal dialysis, which required long term antibiotic therapy, peritoneal dialysis catheter removal, and surgical debridement of the abscess. Nontuberculous mycobacteria should be considered as a possible causative organism for an exit site infection that fails to respond to usual antibiotic therapy. Nontuberculous mycobacterial exit site infections may require peritoneal dialysis catheter removal and surgical debridement. PMID- 15880271 TI - Neonatal transient renal failure with renal medullary hyperechogenicity: clinical and laboratory features. AB - Sonographic findings of renal medullary hyperechogenicity have been observed in the neonate in association with severe perinatal renal injury, kidney malformations or nephrocalcinosis, and, rarely, in newborn infants with transient renal failure. The aim of the study was to describe the entity of neonatal transient renal failure with renal medullary hyperechogenicity (NTRFMH). We studied nine term neonates, born between August 1999 and February 2004 in our institution (0.1% of the live born infants), who developed transient renal dysfunction after birth, and in whom renal sonograms showed bilateral medullary hyperechogenicity. Seven of the infants (78%) had anuria until 30-45 hours of age, and two (22%) had oliguria. Peak serum creatinine levels ranged between 0.61 and 1.62 mg/dL (mean: 1.09+/-0.27 mg/dL) at 2-3 days of life. Additional findings included proteinuria in nine infants (100%), uric acid crystalluria in seven (78%), hyperuricemia in four (44%), and hypertension in one (11%). Hyperuricosuria was demonstrated in one out of the seven patients in whom this parameter was determined. Urinary excretion rates of calcium, phosphorus and oxalic acid were normal, as were urinary levels of amino acids and organic acids. Full clinical recovery accompanied by normalization of all laboratory parameters was observed in all infants by 4-6 days of life. Subsequent follow-up showed normal renal function, no urinary abnormalities, and normal renal sonograms in all infants. Our summary of the nine infants with NTRFMH reported on here and a review of 19 cases of this condition reported in the literature reveal a not-so rare entity of unclear etiology, but excellent prognosis. Physicians caring for neonates should be aware of this benign and transient condition. PMID- 15880272 TI - Multicystic dysplastic kidney detected by prenatal ultrasonography: conservative management. PMID- 15880273 TI - Ventricular arrhythmia following short-acting nifedipine administration. AB - Short-acting nifedipine is still advocated for use in children with severe hypertension, but is no longer recommended for use in adults because of adverse effects from rapid blood pressure reduction. A 19 year-old adolescent with symptomatic, severe hypertension (blood pressure 180/120) received 10 mg of short acting nifedipine sublingually for blood pressure reduction. Within minutes after the dose, the patient complained of palpitations. Tachycardia (heart rate 100 beats per minute) and bigeminy were noted on the cardiac monitor. The bigeminy resolved but premature ventricular contractions were noted for the duration of her hospital stay. We hypothesize that reflex sympathetic activation following an abrupt drop in blood pressure may cause arrhythmias because of elevated catecholamine levels. Given this, it may be more appropriate to treat severe hypertension in children with intravenous antihypertensive agents that can be titrated to produce controlled reductions in blood pressure. PMID- 15880274 TI - The effects of cetrimide-chlorhexidine combination on the hepato-pancreatico biliary system. AB - Sclerosing cholangitis is a rare but dreadful complication of liver hydatid surgery. Almost all scolocidal agents are associated clinically or experimentally with this complication. Cetrimide-chlorhexidine combination is the most potent scolocidal agent, and this study was designed to examine its effect on the hepato pancreatico-biliary system. Forty Wistar-albino rats were divided into two groups. 0.5% cetrimide-0.05% chlorhexidine was injected into the biliary tract of study group animals and 0.9% saline ( NaCl ) into the control group through a 3 mm duodenotomy. The animals were sacrificed after 90 days and histopathological examination of the liver, biliary tract, and pancreas was performed 90 days after the biliary injection. The histopathological examination showed that the lesions ranged from focal necrosis to sclerosing cholangitis to liver cirrhosis. Focal necrosis and eosinophilic inflammation were observed in all rats in the study group. The changes in the liver, biliary tract, and pancreas of the study group animals were significantly more severe than those in the control group (p < 0.05). This experimental study confirms the deleterious effect of another scolocidal agents on the hepato-pancreatico-biliary system. Although it is a very effective scolocidal agent, it should not be used in the presence of a duct biliary-cyst communication. PMID- 15880275 TI - Protein C as an early marker of severe septic complications in diffuse secondary peritonitis. AB - To evaluate the predictive value of protein C as a marker of severity in patients with diffuse peritonitis and abdominal sepsis, protein C levels were repeatedly determined and compared with serum levels of antithrombin III, plasminogen, alpha(2)-antiplasmin, Plasminogen activator inhibitor, D-dimer, C1-inhibitor, high molecular weight kininogen, and the C5a, C5b-9 fragments of the complement system. We carried out a prospective study from 44 patients with severe peritonitis confirmed by laparotomy and 15 patients undergoing elective ventral hernia repair who acted as controls. Analyzed biochemical parameters were determined before operations and on days 1, 2, 3, 5, 7, 10, and 14 after operations. For the study group, preoperative average protein C level was significantly lower in the patients who developed septic shock in the late course of the disease, with lethal outcome, than in the patients with severe peritonitis and sepsis who survived (p = 0.0001). In non-survivors, protein C activity remained decreased below 70%, whereas the course of survivors was characterized by increased values that were significantly higher (p < 0.03) at every time point than in those patients who died. Protein C was of excellent predictive value and achieved a sensitivity of 80% and a specificity of 87.5% in discriminating survivors from non-survivors within the first 48 hours of the study (AUC-0.917; p < 0.001), with a "cut-off" level of 66.0%. As for the control group, throughout the study period, protein C activity was permanently maintained within the range of normal, with significant differences with reference to the study group (p < 0.01). These results suggest that protein C represents a sensitive and early marker for the prediction of severe septic complications during diffuse peritonitis, and of outcome. PMID- 15880276 TI - Number of lymph node metastases is a significant prognostic factor in intrahepatic cholangiocarcinoma. AB - Intrahepatic cholangiocarcinoma (IHCC) is a rare primary hepatic tumor. Outcomes after resection and the use of lymph node dissection have not been well described. From a prospective database, we identified 53 patients with IHCC who underwent exploration between April 1983 and March 2004. Hepatic resection was performed in 44 patients, 30 of whom underwent lymph node dissection. Clinicopathological features and outcomes were analyzed. The actuarial 1-year survival was 66.2% in resected patients, compared to 0% in unresectable patients (p < 0.0001), with a 50% overall survival of 21.5 months and 3.1 months, respectively. The actuarial 3-year and 5-year overall survival rates in resected patients were 38.3% and 26.3%, respectively. Univariate analysis revealed that factors associated with poor overall survival included multiple tumors, extrahepatic bile duct involvement, noncurative resection, and involvement of lymph nodes. Multivariate analysis in resected patients revealed that multiple tumors (p < 0.0074) and non-curative resection (p = 0.0068) were significant risk factors for poor overall survival. The survival rate in patients with three or more positive nodes was significantly lower than in those with fewer than three (p < 0.0001). Three patients with solitary tumors and one or two involved lymph nodes have survived beyond 4 years after extended lobectomy with systemic lymphadenectomy. Curative resection, single tumor, and fewer than two lymph node metastases were prognostic factors for good outcome. Curative resection with lymph node dissection improved survival in patients with no more than two positive lymph nodes. PMID- 15880277 TI - Eleven years of liver trauma: the Scottish experience. AB - The aim of this population based study was to assess the incidence, mechanisms, management, and outcome of patients who sustained hepatic trauma in Scotland (population 5 million) over the period 1992-2002. The Scottish Trauma Audit Group database was searched for details of any patient with liver trauma. Data on identified patients were analyzed for demographic information, mechanisms of injury, associated injuries, hemodynamic stability on presentation, management, and outcome. A total of 783 patients were identified as having sustained liver trauma. The male-to-female ratio was 3:1 with a median age of 31 years. Blunt trauma (especially road traffic accidents) accounted for 69% of injuries. Liver trauma was associated with injuries to the chest, head, and abdominal injuries other than liver injury; most commonly spleen and kidneys. In all, 166 patients died in the emergency department, and a further 164 died in hospital. The mortality rate was higher in patients with increasing age (p < 0.001), hemodynamic instability (p < 0.001), blunt trauma (p < 0.001), and increasing severity of liver injury (p < 0.001). The incidence of liver trauma in Scotland is low, but it accounts for significant mortality. Associated injuries were common. Outcome was worse in patients with advanced age, blunt trauma, multiple injuries and those requiring an immediate laparotomy. PMID- 15880278 TI - Indication of hepatectomy for cirrhotic patients with hepatocellular carcinoma classified as Child-Pugh class B. AB - We clarified the indication of partial hepatectomy in hepatocellular carcinoma (HCC) patients with liver cirrhosis classified as Child-Pugh class B. Univariate analysis revealed that adverse prognostic factors were (1) the presence of ascites, (2) elevated total bilirubin (1.5 mg/dl or higher), (3) reduced choline esterase (160 IU/ or lower), (4) elevated alpha-fetoprotein (AFP) (400 ng/ml or higher), (5) microscopic vascular invasion, and (6) non-curative hepatectomy. Microvascular invasion was excluded in the multivariate analysis because this factor could not be predicted before hepatectomy. Multivariate analysis revealed that independent adverse prognostic factors were (1) elevated total bilirubin (1.5 mg/dl or higher), (2) presence of ascites, (3) elevated AFP (400 ng/ml or higher), and (4) non-curative hepatectomy. The overall 5-year survival rate of patients with none of or only one of the four adverse prognostic factors was 45.8%. The overall 5-year survival rate of patients with two or more adverse prognostic factors was only 7.0%. Partial hepatectomy is the first choice of treatment for patients with none of or only one of the four adverse prognostic factors, whereas orthotopic liver transplantation or other conservative treatment should be considered for patients with two or more adverse prognostic factors. PMID- 15880279 TI - Malignant metastatic insulinoma-postoperative treatment and follow-up. AB - The rarity of malignant insulinoma limits reports on therapeutic strategies and outcome. The treatment and follow-up of 10 patients, all presenting an insulinoma with metastatic disease of the liver and newly diagnosed between 1992 and 2002, is reported. Pancreatic surgery with successful removal of the primary tumor preferentially located in the tail was performed in 7 women and 3 men, median age 55 years (range 36-82 years). If appropriate, 5 patients underwent additional hepatic surgery and lymph node resections. Liver metastases as the major cause of postoperatively persistent hypoglycemia were subsequently treated by repeated transarterial hepatic chemoembolization and chemoperfusion protocols using high dose transhepatic streptozocin perfusions (3-4 g per session). The current median survival time for all 10 patients is 2.6 years (range: 1.6-9.7 years). Six patients are currently alive with a median survival of 3.7 years (1.7-9.7 years), five of them with stable disease and free of hypoglycemia. Four patients died after a median survival of 1.8 years (range: 1.6-7.5 years) from complications of unmanageable hypoglycemia. It is concluded that the necessity to treat debiliating and life-threatening hypoglycemia in metastatic malignant insulinoma warrants the option of radical endocrine surgery in combination with extended and repeated postoperative chemoembolization of liver metastases. PMID- 15880280 TI - Effect of radiofrequency ablation of the liver on cell-mediated immunity in rats. AB - We investigated the effect of radiofrequency ablation (RFA) of the liver on cell mediated immunity in rats. Sprague-Dawley rats were divided into five groups: control group that did not have any procedure, a sham-operation group that underwent laparotomy, a lobectomy group that had left lateral lobectomy of the liver, a one-lobe RFA group that had RFA on the left lateral lobe of the liver, and a two-lobe RFA group that had RFA on the left lateral and left median lobe of the liver. Delayed-type hypersensitivity (DTH) was induced by challenging the ears of previously sensitized rats with 2,4-dinitro-1-fluorobenzene. Cell mediated immunity was evaluated by direct measurement of the ear thickness and quantitative analysis of leukocytic infiltration of the tissue. The increased ear thickness was 42.0%, 69.2%, 61.8%, 46.7%, and 39.8% from the baseline one day after each procedure, and the mean leukocytic infiltration into the ear tissue was 3136.7, 3895.0, 3913.3, 2803.3, and 2316.7/mm(2) seven days after each procedure in the control, sham-operation, lobectomy, one-lobe RFA, and two-lobe RFA groups, respectively. In conclusion, RFA of the liver tissue can partially abolish the augmentation of DTH and leukocytic infiltration those were seen in the hepatic lobectomy and sham-operation groups. The procedure had a negative effect on cell-mediated immunity in rats, and the intensity of RFA showed a reverse correlation with cell-mediated immunity. PMID- 15880281 TI - Safety and effectiveness of left hepatic trisegmentectomy for hilar cholangiocarcinoma. AB - Left hepatic trisegmentectomy has been performed for huge malignant tumors, but it is rarely applied in patients with hilar cholangiocarcinoma. Twelve consecutive patients (7 men and 5 women; mean age, 64 years) underwent left hepatic trisegmentectomy in our institution between January 2000 and December 2003. The preoperative management and postoperative outcomes of this surgical procedure were presented and retrospectively analyzed. Preoperative biliary drainage and portal vein embolization were performed in 6 patients (50%) and 9 patients (75%), respectively. The preoperative estimated volume ratio of the posterior segment /the whole liver was 44.8 +/- 7.0% (34.3-54.3), the plasma retention rate of indocyanine green at 15 minutes was 8.6 +/- 2.2% (4.7-13.7), and the serum total bilirubin level before surgery was 1.0 +/- 0.4 mg/dl (0.4 1.7). The serum total bilirubin level on the first postoperative day was 3.3 +/- 0.4 mg/dl (1.4-6.2). There was no hospital death or postoperative hepatic failure. The incidence of positive resectional margin was 25%. With biliary decompression and preoperative portal embolization techniques, left hepatic trisegmentectomy was a safe and curative resectional option for hilar cholangiocarcinoma. PMID- 15880282 TI - Preoperative albendazole and scolices viability in patients with hepatic echinococcosis. AB - To determine the plasmatic and intracystal concentrations of albendazole sulfoxide (AS) and correlate them with the viability of the scolices in patients surgically treated for hepatic hydatid cysts (HHC) that received albendazole preoperatively, as an indirect way of evaluating the scolicide efficacy of the drug. A non-consecutive series of patients with uncomplicated HHC, underwent operation at the Department of Surgery, Regional Hospital of Temuco, Chile, between 2001 and 2002. The patients were given 10 mg/kg/day of albendazole for 4 days prior to the surgery. Intraoperative samples of venous blood and hydatid fluid were taken, in which the plasmatic concentration (PIC) and intracystal concentration (ICC) of AS were measured by means of high-performance liquid chromatography. With the remaining hydatid fluid, the viability of the scolices was examined. The following variables were taken into consideration: diameter, type of cyst, number of cysts, and development of cyst-biliary communications. Descriptive statistics were used in the calculation of medians, averages, and standard deviations, and analytical statistics were used for the comparison of continuous variables applying the t-test and the Mann-Whitney U-test. A total of 26 patients with HHC, with a median age of 39.5 years (range: 16-80 years); 16 were women (61.5%). Nineteen patients presented only one cyst (73.1%), and the diametric median of the cysts was 14.5 cm (range: 7-30). Fourteen patients presented univesicular cysts (53.9%), and the remaining 12 had multivesicular cysts (46.1%). The average PIC of AS in the whole series was 1.88 +/- 0.5 microg/ml and the ICC was 0.26 +/- 0.2 microg/ml. The ICC in viable hydatid cysts was 0.25 microg/ml versus 0.28 microg/ml in non-viable cysts (p = 0.7556). The absence of association between intracystal levels of AS and the viability of the scolices allows one to posit indirectly that albendazole is ineffective as a scolicidal agent administered preoperatively for 4 days. PMID- 15880283 TI - Adult surgical emergencies in a developing country: the experience of Nnamdi Azikiwe University Teaching Hospital, Nnewi, Anambra State, Nigeria. AB - The goal of this study was to examine the adult surgical emergencies seen at the Nnamdi Azikiwe University Teaching Hospital (NAUTH), Nnewi, with a view to proffering preventive solutions where appropriate and improving outcome. From the register of patients seen at the Casualty department and from the operations register in the main operation room of NAUTH, names and hospital numbers of adult patients treated as emergencies over a 5-year period, from 7 September 1998 to 6 September 2003, were obtained. The hospital folders were then retrieved from the Records Department. From each folder, the following details about each patient were extracted: age, sex, diagnosis at presentation, causative factors, treatment given, and outcome. A total of 902 adult patients were treated during the period. The commonest emergency operation was appendectomy for acute appendicitis in 139 patients (97 women and 42 men), followed closely by road traffic accidents (RTAs) involving 137 patients (103 men and 34 women). Gunshot injuries, which resulted mainly from armed robbery attacks, accounted for 127 cases. More men (113) sustained gunshot injuries than women (14). Of the 92 cases of acute intestinal obstruction seen, 62 occurred in women and 30 in men. Some 126 men presented with acute urinary retention, and two others presented with priapism. Governments at various levels should provide modern diagnostic tools for the accurate preoperative diagnosis of surgical emergencies in hospitals. Governments should also inculcate strict discipline into drivers using the highways, particularly in relation to abuse of alcohol and drugs. Good roads and adequate security should be provided for the people. The need for Pre-Hospital Care for the efficient evacuation of accident victims is emphasized. These measures will help to improve the management and outcome of surgical emergencies, and decrease the number of surgical emergencies resulting from RTAs and gunshot wounds. PMID- 15880286 TI - Laparoscopic telementored adrenalectomy: the Italian experience. AB - BACKGROUND: Laparoscopy is widely accepted as the gold standard for adrenalectomy. Telementoring has been developed to reduce the complications associated with surgeon inexperience. We report our preliminary experience with laparoscopic telementored adrenalectomy. METHODS: From July 2002 to May 2003, eight laparoscopic telementored adrenalectomies were performed between two separate operating sites 430 km apart. Six of these procedures were monolateral laparoscopic adrenalectomies, and one was bilateral. All cases were performed by an expert open surgeon who was skilled in laparoscopic procedure but who had no experience in laparascopic adrenalectomy RESULTS: All the procedures were successfully performed in a telementored fashion. The mean operative times, blood loss, and postoperative morbidity results were comparable to those for standard laparoscopic adrenalectomies reported in the literature. CONCLUSIONS: This preliminary experience has demonstrated the feasibility of national telementoring. It is a viable method that can potentially add to surgical education and decrease the likelihood of complications due to inexperience with new techniques. PMID- 15880287 TI - Costs and quality of life after endoscopic repair of inguinal hernia vs open tension-free repair: a review. AB - BACKGROUND: The ongoing debate about the relative merits of endoscopic (EH) vs open mesh herniorrhaphy (OH) prompts the need for comparisons of outcome measures other than recurrence. Therefore, we reviewed data on the costs, time to return to work, quality of life (QoL), and pain associated with EH and OH. METHODS: Studies comparing EH to OH and explicitly involving costs or QoL were identified and reviewed. RESULTS: Eighteen studies were included. Direct in-hospital costs were higher for unilateral EH. Direct out-of-hospital costs were lower after EH in some studies. Indirect costs were lower for EH. Total costs were higher for EH in three studies and lower in one study. With EH, QoL was better, pain was less, operating time was longer, and time return to work and other activities was shorter. CONCLUSION: From a societal perspective, EH entails costs similar to OH but offers extra benefits to the patient in terms of QoL and pain. PMID- 15880288 TI - The role of heat-conducting walls in the measurement of heat and mass transport in transient grating experiments. AB - We present a two-dimensional model to account for the role of heat-conducting walls in the measurement of heat transport and Soret-effect--driven mass transport in transient holographic grating experiments. Heat diffusion into the walls leads to non-exponential decay of the temperature grating. Under certain experimental conditions it can be approximated by an exponential function and assigned an apparent thermal diffusivity D(th, app)20 gL(-1)) form transient gels exhibiting an oscillatory shear behavior that can be approximated by a single-relaxation Maxwellian model. Aqueous solutions of C(18)-PNIPAM-C(18) undergo a phase transition upon heating to 31.5( degrees )C as determined by microcalorimetry. The heat-induced phase separation of dilute (0.025 gL(-1)) C(18)-PNIPAM-C(18) solutions yields a fluid that is colloidally stable at temperatures higher than 33( degrees )C. The overall results are consistent with a model assuming the formation of flowerlike micelles in the dilute regime and a network of micelles connected by telechelic chains in the concentrated regime. PMID- 15880291 TI - The bubble cloud as an N-degree of freedom harmonic oscillator. AB - The dynamics of a two-dimensional N-bubble static cloud is investigated and shown to be well described by an N-degree of freedom harmonic oscillator model, at least at low enough frequencies. Eigenmodes and eigenfrequencies are calculated and compared with experimental results obtained with an assembly of bubbles caught up under a net in a water tank. Accordance is found to be excellent in the frequency range of validity of the model, the limits of which are discussed. An interpretation of the low-frequency branch of Foldy's dispersion relation in bubbly liquids is suggested in terms of "bubble waves" in a quasi-incompressible medium. PMID- 15880292 TI - Treatment of urethral stricture in a female-to-male transsexual with a tubulized flap from the labia minora. AB - Urethral strictures are not uncommon after microvascular phalloplasty in female to-male transsexuals, and usually are secondary to technical errors or vascular complications. A case of a female-to-male transsexual with recalcitrant urethral stricture after microvascular phalloplasty is reported. After one unsuccessful attempt at direct repair and a failed grafting procedure, a tubulized island flap from the remnants of the labia minora was successfully used to reconstruct the urethral stenosis. Current options for treatment of urethral stenoses in female to-male transsexuals are discussed. PMID- 15880293 TI - Salvage of an ischemic "kite" flap by arteriovenous supercharging. AB - The neurovascular "kite" flap is a well-established procedure for single-stage reconstruction of soft-tissue defects in the thumb. Post-surgical ischemia of the flap due to poor inflow is a rare, but potential, complication. The authors describe a technique of arteriovenous supercharging that was successfully used to salvage such an ischemic flap. PMID- 15880294 TI - Distraction lengthening of the thumb following replantation. AB - Distraction lengthening is one of several surgical reconstructive methods used to obtain length of an amputated digit and to improve function. However, the use of this technique following replantation is a relatively new concept. A case of a 32 year-old male who sustained a traumatic amputation of his left nondominant thumb and index finger is described. The injury and the replantation surgery each resulted in a significant amount of bone loss from the thumb. Nine months following the replantation, distraction osteogenesis was utilized to lengthen the thumb and to improve both function and cosmesis. PMID- 15880295 TI - Flow-through, functioning, free musculocutaneous flap transfer for restoration of a mangled extremity. AB - Reconstructing a mangled extremity requires careful, complete examination of the functional and structural status of the injured part. Although the vascular status of the injured extremity is a priority, simultaneous reconstruction of skeletal, vascular, and neuromuscular structures should be considered in a combined approach. The purpose of this study was to describe a one-stage reconstruction of both soft-tissue coverage and revascularization for a lower extremity, along with functional restoration of an anterior lower leg. For this purpose, the functioning transfer of a free latissimus dorsi musculocutaneous flap as a flow-through type was performed. The results demonstrate that the technique is extremely useful in limb-salvage procedures from both functional and structural viewpoints. PMID- 15880296 TI - A pedicle-lengthening technique for free latissimus dorsi muscle flaps: the "Y-V I" principle. AB - This article describes a pedicle-lengthening procedure (the Y-V-I principle) that reverts a Y-shaped vascular structure into an I-shaped vascular structure, and its application to the latissimus dorsi muscle. If the Y-shaped vascular structure is divided proximal to the branching point and ligated, it becomes a V shaped vascular structure. To obtain a linear I-shaped vascular structure, the V is opened, and the length of the vascular pedicle will become equal to the combined length of both limbs of the V. This report indicates that the latissimus dorsi flap may be transferred based on the circumflex scapular artery or the serratus branch. PMID- 15880297 TI - Genes and nerves. AB - The unpredictability of a brachial plexus graft, a median nerve repair, or a facial-nerve reconstruction is well known. No matter how precise the technical skills, a perfect recovery from a peripheral-nerve lesion is elusive. To resolve this problem, understanding of the normal development of the peripheral nervous system is needed. Presently, the development of the innervation in the upper limb is complex and not fully understood. However, many of the genes involved in this process are now known, and the link between anatomy and genetics is becoming clearer. This short review aims to acquaint the clinical surgeon with some of the main genes. The principal steps in the establishment of neural circuits will be summarized, in particular, the specification and development of neurons and glia, the pathfinding of cells and axons towards their target, and the downstream molecules that control the circuitry of these neurons. PMID- 15880298 TI - Pectoral skin flap as a reliable and simple model for vascularized composite skin transplantation research. AB - In this study, the authors explored the feasibility of transplanting a "pectoral skin flap" as an alternative model for composite skin allotransplantation research. Genetically inbred male Lewis rats (LEW; RT1l) (n = 13), weighing 250 to 300 gr, were used as recipients and Brown Norway rats (BN; RT1n) (n = 8) rats were used as donors. Five Brown Norway rats had bilateral and three had unilateral flap harvest with a total of 13 flap transplantations to Lewis rats. All flaps were transplanted to the groin regions of the recipients and microsurgical anastomoses with 10-0 nylon were performed between the axillary vessels of the flaps and the femoral vessels of the recipient animals. The mean operation time was 55 min, with an ischemia time of 25 min. All transplantations were successful immediately postoperatively. The recipient animals were treated with Cyclosporine A (16 mg/kg/day) from the day of the surgery for 1 week, and then discontinued. One flap developed infection at postoperative day 11 and one flap had partial (50 percent) necrosis. The early signs of rejection started 8 to 9 days after the cessation of the immunosuppressive treatment, and all flaps were acutely and uniformly rejected within 3 to 4 days. The feasibility of harvesting two flaps from one donor animal allowed saving donor animal lives and reducing donor-specific variables. PMID- 15880299 TI - Evaluation of fructose 1,6 diphosphate for salvage of ischemic gracilis flaps in rats. AB - Fructose 1, 6 diphosphate (FDP), a metabolic intermediate, provides an alternative mechanism to circumvent the rate-limiting step in the Kreb's cycle. This agent has been observed to prevent the effects of ischemia on heart tissue and kidney function and the effects of endotoxic shock. It has been shown conclusively to minimize the adverse effects of ischemia-reperfusion injury in experimental pedicled skin flaps in animals. The present study was done to evaluate the effect of intra-arterial administration of FDP on salvage of ischemic microvascular transfer of gracilis muscle flaps in rats, with the premise that it might prolong the ischemia time of muscle flaps at room temperature, thus increasing chances of flap survival. Irrigation with FDP did not change the quantitative survival of the flaps, but there was qualitative improvement on histologic evaluation and DNA analysis. Decreased inflammatory damage and DNA fragmentation were seen at the 2.5-hr period. Histologic staining for mitochondrial oxygenation in gracilis muscle also showed increased uptake in the FDP-treated group vs. control at the 2.5-hr ischemia period. Further experiments with different modes of FDP administration should be carried out to identify more effective means of amelioration of flap ischemia. PMID- 15880300 TI - Overcoming peripheral nerve gap defects using an intact nerve bridge in a rabbit model. AB - This study investigated the intact nerve bridge technique for overcoming peripheral nerve gap defects in a rabbit model. To create the intact nerve bridge, a 1-cm segment of the peroneal nerve is resected leaving a gap defect. The proximal and distal peroneal nerve stumps are sutured 1 cm apart, in an end to-side fashion, to the intact tibial nerve epineurium. Four experimental groups were used (n = 10): primary repair of resected segment; intact nerve bridge; nerve autograft; and gap in situ control. Evaluation after 12 weeks included measurement of isometric muscle contraction force, axonal counting, wet muscle weights, and histologic examination. The results of this study support two main conclusions, in a rabbit model: (a) regenerating axons can use the epineurium of an intact nerve to bridge a gap defect; (b) there is no significant difference in the functional recovery between standard nerve autografts and the intact nerve bridge technique. PMID- 15880301 TI - Peripheral nerve transplantation: the role of chemical acellularization in eliminating allograft antigenicity. AB - This study tests the hypothesis that a chemically acellularized peripheral nerve allograft is as immunologically inactive as a peripheral nerve isograft. Cellular and acellular sciatic nerves were transplanted from BALB/c into C57BL/6 mice. C57BL/6 sciatic nerves were also transplanted into C57BL/6 recipients as isograft controls. Fourteen days post-transplantation, recipient splenocytes were isolated, stimulated with donor alloantigens, and IL-2, IL-4, IL-5, and gamma-IFN production was quantified using the ELISPOT technique. Cellular peripheral nerve allografts stimulated robust Th1 and Th2 systemic immune responses, whereas acellular peripheral nerve allografts elicited a response that is comparable to or lower than that quantified following peripheral nerve isograft transplantation. Chemical acellularization of peripheral nerve allografts dramatically reduces the cellular and humoral immunologic responses. These data indicate that chemically acellularized peripheral nerve constructs are relatively non-antigenic and may be a readily available source of nerve for peripheral nerve reconstruction. PMID- 15880302 TI - [Situation and context appropriateness of guidelines]. PMID- 15880303 TI - [Guidelines and the psychology of medical decision making in the treatment of depressive disorders]. AB - Guidelines aim at improving medical decision making. Guidelines try to influence medical behaviour in contrast to textbooks and reviews which want to improve medical knowledge. Scientific models of medical decision making such as the action theory and empirical data on the effects of guidelines suggest that guidelines will not always reach their goal but instead can even deteriorate medical quality. The problem of guidelines in medicine is discussed in reference to depressive disorders. The problems at stake are nevertheless valid for medicine as a whole. Therefore it must be tested in controlled clinical trials whether guideline exposed physicians have better results with their patients than guideline naive physicians. Only such guidelines should be called evidence based for which positive effects have been empirically demonstrated. PMID- 15880304 TI - [Pharmacotherapy in personality disorders]. AB - Subjects with personality disorders represent a relevant subset of the clinical psychiatric population and, to an even bigger extent, a comorbid condition to an axis-I diagnosis. Personality disorders appear to be difficult to treat and problematic in the therapeutic setting, and a pharmacologic treatment often does not follow recommendations from empirical studies. Psychopharmacologic drugs are able to modify the expression of certain personality dimensions, which are mainly disorder-related variables of temperament. Modern pharmacotherapy of personality disorders has a neurobiological basis. This is underlined by evidence from animal studies and can be associated with certain receptor-specific functions in humans. Treatment is consequently symptom-related and not specific for any disorder. It must be stressed that certain features but not the entire personality can be modified by drug treatment. Data from drug studies are still relatively scarce. An overview of studies on personality disorders is presented including information on dosages and augmenting strategies. Psychotherapy still is the basic treatment for personality disorders. However, psychopharmacological interventions should not only be considered when psychotherapy has failed. Treatments should be complementary from the start. There is a rationale for short and long-term treatment. The latter is indicated, despite of the scarcity of data, when longer lasting and severe affective symptoms emerge. Response rates vary among the trials and appear to be lower than in non-comorbid axis-I disorders. A treatment algorithm is presented. PMID- 15880305 TI - [Therapy of pain syndromes in multiple sclerosis -- an overview with evidence based recommendations]. AB - While pain is a common problem in multiple sclerosis (MS) patients, it is frequently overlooked and has to be asked for actively. Pain can be classified into 4 diagnostically and therapeutically relevant categories. 1. PAIN DIRECTLY RELATED TO MS: Painful paroxysmal symptoms like trigeminal neuralgia or painful tonic spasms are treated with carbamazepine as first choice, or lamotrigine, gabapentin, oxcarbazepine and other anticonvulsants. Painful "burning" dysaesthesia, the most frequent chronic pain syndrome, are treated with tricyclic antidepressants or carbamazepine, further options include gabapentin or lamotrigine. While escalation therapy may require opioids, the role of cannabinoids in the treatment of pain still has to be determined. 2. PAIN INDIRECTLY RELATED TO MS: Pain related to spasticity often improves with adequate physiotherapy. Drug treatment includes antispastic agents like baclofen or tizanidine, alternatively gabapentin. In severe cases botulinum toxin injections or intrathecal baclofen merit consideration. Physiotherapy and physical therapy may ameliorate malposition-induced joint and muscle pain. Moreover, painful pressure lesions should be avoided using optimally adjusted aids. 3. Treatment related pain can occur with subcutaneous injections of beta interferons or glatiramer acetate and may be reduced by optimizing the injection technique and by local cooling. Systemic side effects of interferons like myalgias can be reduced by paracetamol or ibuprofen. 4. Pain unrelated to MS such as back pain or headache are frequent in MS patients and may be worsened by the disease. Treatment should be follow established guidelines. In summary, a careful analysis of the pain syndrome will allow the design of the appropriate treatment plan using various medical and non-medical options and thus will help to ameliorate the patients' quality of life. PMID- 15880306 TI - [Ataxia]. PMID- 15880307 TI - Endoscopic retrieval of a small bowel enteroscopy capsule lodged in a Zenker's diverticulum. PMID- 15880308 TI - From bases to basis: linking genetics to causation in primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a multifactorial autoimmune disease with inherited and environmental components in pathogenesis. It is exceptional among autoimmune diseases in showing strong heritability according to familial occurrence and monozygotic twins concordance, yet with weak associations with the usual genetic risk elements for autoimmunity, such as the HLA alleles. Among the latter, there is risk (at least in some populations) conferred by HLA DRB1*08 and possibly some protection by DRB1*11. However, the inconsistency among studies on HLA is surprising, given that PBC is a relatively homogenous disease entity. Among non-HLA genes, some studies implicate polymorphisms of genes for cytotoxic T-lymphocyte antigen-4, interleukin-2, or interleukin-10; polymorphisms of the vitamin D receptor could synergize with low sunlight exposure to create deficiency of the immunoregulatory factor, activated vitamin D. A new lead is available from the finding in female subjects with PBC of an increase in the degree of monosomy of the X chromosome that is presumed to carry immune response genes. A further suggested source of inquiry is the apparent protection of African-American women from PBC. Finally, data on inheritance should be sought in PBC by descent methodology, rather than by cross-sectional association studies in cases and control subjects, and based on analysis of a large number of families with an affected member through a worldwide effort. PMID- 15880309 TI - Capsule endoscopy: practical applications. AB - Few advances in the history of gastroenterology have made as dramatic an impact on the diagnosis of gastrointestinal disease as the development and rapid clinical implementation of wireless capsule endoscopy. Less than 4 years after the landmark publication, capsule endoscopy is widely considered an essential component of the diagnostic workup of obscure gastrointestinal bleeding, and its role is expanding in the diagnosis of small bowel diseases such as Crohn's disease. This review appraises the available literature and highlights practical aspects of capsule endoscopy of interest to the clinician. We discuss safety concerns, optimal preparation for the procedure, diagnostic utility as compared to conventional methods, indications for capsule endoscopy, and outcomes. PMID- 15880310 TI - Multiple endocrine neoplasia type 2B and Hirschsprung's disease. AB - Multiple endocrine neoplasia type 2B and Hirschsprung's disease are genetic disorders characterized by gross and/or microscopic pathology of the enteric nervous system and associated dysmotility. A specific missense mutation in the RET proto-oncogene is the etiology of multiple endocrine neoplasia type B, in contrast to very complex multigenetic defects that underlie Hirschsprung's disease, which include overt mutations and more subtle changes in the RET locus. In this review, the molecular genetics of the 2 conditions are discussed, and the clinical implications of existing data and future studies are summarized. PMID- 15880311 TI - Severe constipation. PMID- 15880312 TI - Prevalence of functional gastrointestinal disorders in women who report domestic violence to the police. AB - BACKGROUND & AIMS: Retrospective studies found an association between past sexual, physical, or psychological abuse and functional gastrointestinal disorders (FGIDs). However, there are no studies evaluating such an association concurrently with the ongoing abuse. Our aim was to investigate the prevalence of the main FGIDs, functional dyspepsia and irritable bowel syndrome, in 70 women reporting a situation of domestic violence to the police and to evaluate the level of psychological distress and its relationship with the presence of FGID. METHODS: Through an interview between a social worker and the woman reporting abuse, digestive symptoms, psychological status, and type of abuse were recorded. These data were matched against police records. Functional dyspepsia and irritable bowel syndrome were diagnosed according to Rome II criteria. RESULTS: Seventy-one percent of the women had an FGID: 67% functional dyspepsia, 47% irritable bowel syndrome, and 43% both. In two thirds of the cases, FGID onset occurred simultaneously with or soon after abuse onset. Only 34% of the women had sought medical attention for FGID symptoms. No differences were found between women with or without FGID regarding age and type or duration of abuse; psychological distress tended to be more severe in the group of women with FGIDs. CONCLUSIONS: Most women who suffer domestic violence (reported to the police) have functional dyspepsia and/or irritable bowel syndrome and also have elevated psychological distress. This has important implications, not only for comprehensive health care of women in a situation of abuse, but also for medical treatment of women with FGIDs. PMID- 15880313 TI - Effect of the prebiotic oligofructose on relapse of Clostridium difficile associated diarrhea: a randomized, controlled study. AB - BACKGROUND & AIMS: Ten percent to 20% of patients relapse after successful treatment of their Clostridium difficile -associated diarrhea. We set out to determine if the prebiotic oligofructose could alter the fecal bacterial flora and, in addition to antibiotic treatment, reduce the rate of relapse from C difficile infection. METHODS: Consecutive inpatients with C difficile -associated diarrhea were randomly allocated to receive oligofructose or placebo for 30 days in addition to specific antibiotic treatment. Patients were followed up for an additional 30 days. The main end point was the development of further diarrhea. Stools were collected for bacterial culture and C difficile toxin measurement. RESULTS: One hundred forty-two patients were recruited. Stool culture confirmed the probiotic effect of oligofructose with an increase in fecal bifidobacteria from baseline 8.68 log(10) colony-forming units (cfu)/g to 9.37 log(10) cfu/g at discharge (P < .0001; 95% confidence interval [CI], 0.45-0.94), 9.64 log(10) cfu/g at 30 days (P < .0001; 95% CI, 0.74-1.18), and 9.42 log(10) cfu/g at 60 days (P < .0001; 95% CI, 0.56-0.93). Thirty patients experienced a relapse of diarrhea after a median of 18 days (range, 8-34 days). Relapse of diarrhea was more common in those taking placebo (8.3% oligofructose vs 34.3% placebo, P < .001, chi(2) = 14.35). Patients who relapsed stayed in the hospital longer than those who did not (53 vs 26 days, P = .021; 95% CI, 2-28), and there was a longer period of time from commencing metronidazole or vancomycin and their diarrhea settling (6 vs 3 days; P = .007; 95% CI, 1.0-5.0). CONCLUSIONS: Fecal cultures confirmed the prebiotic effect of oligofructose. Patients taking oligofructose were less likely to develop further diarrhea than those taking the placebo. PMID- 15880314 TI - Natural history of chronic idiopathic intestinal pseudo-obstruction in adults: a single center study. AB - BACKGROUND & AIMS: Chronic idiopathic intestinal pseudo-obstruction (CIIP) is a rare disease characterized by episodes resembling mechanical obstruction in the absence of organic, systemic, or metabolic disorders. Intestinal motor abnormalities have long been identified in CIIP patients. Little is known of the natural history of the disease in adults. This study evaluated the clinical course of CIIP over time. METHODS: Fifty-nine consecutive CIIP patients without underlying collagen, vascular diseases, or mitochondrial cytopathies were evaluated between 1985 and 2001. Family history, onset of digestive symptoms, previous surgeries, episodes suggestive of subacute intestinal obstruction, digestive symptoms, body mass index, and feeding habits were recorded. Small bowel manometry was performed by a perfusion technique, and abnormal motor patterns were visually identified. Full-thickness biopsies were available in 11 cases and were processed for immunohistochemical analysis of myogenic and neurogenic components of the gut wall. RESULTS: Patients were prospectively followed up for a median of 4.6 years (range, 1-13 years). Diagnosis was often made several years after symptom onset (median, 8 years). Thus, the majority of patients (88%) underwent useless and potentially dangerous surgeries (mean, 2.96 per patient). Manometry invariably showed abnormal motor patterns. Pathologic findings included neuropathies in all investigated cases and abnormalities of interstitial cells of Cajal in 5 of 11 cases. Long-term outcome was generally poor despite surgical and medical therapies; 4 patients died of disease-related complications, 4 underwent small bowel transplantation, almost one third required long-term home parenteral nutrition, and two thirds had some sort of nutritional limitations. CONCLUSIONS: CIIP is a severe, often unrecognized disease characterized by disabling and potentially life-threatening complications over time. PMID- 15880315 TI - Antibiotic use and the risk of flare of inflammatory bowel disease. AB - BACKGROUND & AIMS: Intestinal microbial flora participate in the pathogenesis of inflammatory bowel disease. Because antibiotic therapy alters intestinal microbial flora, we hypothesized that use of antibiotics might decrease the risk of flare. METHODS: We conducted a case-crossover study by using the General Practice Research Database from 1989-1997. Flares of disease were identified by receipt of a new prescription for either corticosteroids or mesalamine medications after an interval of at least 4 months without prescriptions for either class of medication. The primary exposure was receipt of any antibiotics in the 60 days preceding the index date. RESULTS: Among 1205 patients with Crohn's disease, exposure to antibiotics was associated with a reduced risk of flare (adjusted odds ratio [OR], 0.78; 95% confidence interval [CI], 0.64-0.96; P = .019). The effect was strongest with more recent exposure (test for trend, P < .05). Among 2230 patients with ulcerative colitis, use of any antibiotics within 60 days was not associated with flare of disease (adjusted OR, 0.96; 95% CI, 0.82 1.12; P = .581), although a potentially protective effect was observed in those patients with very recent exposure (exposure within 15 days: OR, 0.66; 95% CI, 0.51-0.85). CONCLUSIONS: Antibiotic use within 60 days was associated with a lower risk of flare of Crohn's disease, but not ulcerative colitis. The strength of the protective effect of antibiotics in Crohn's disease wanes over time. PMID- 15880316 TI - Supportive nutrition on recovery of metabolism, nutritional state, health-related quality of life, and exercise capacity after major surgery: a randomized study. AB - BACKGROUND & AIMS: The aim of this study was to investigate whether specialized supportive enteral and parenteral feeding have superior effects compared to oral nutrition on recovery during long-term postoperative treatment of cancer patients with preoperative weight loss and reduced maximum exercise capacity. METHODS: One hundred twenty-six patients referred for resection of the esophagus (n = 48), stomach (n = 28), or pancreas (n = 50) were considered to be included before operation. Included patients (n = 80) received supportive enteral or parenteral nutrition postoperatively at home corresponding to 1000 kcal/d until the patients did not wish to continue with artificial nutrition for any reason. Patients randomized to oral nutrition only served as control subjects. Caloric intake, body composition (dual-energy x-ray absorptiometry), and respiratory gas exchanges at rest and during exercise were measured including health-related quality of life. RESULTS: Survival and hospital stay did not differ among the groups, whereas overall complications were higher on artificial nutrition (P < .05). Changes in resting energy expenditure and biochemical tests did not differ during follow-up among the groups. Body weight and whole body fat declined similarly over time in all groups (P < .005), whereas lean body mass was unchanged during follow-up compared to preoperative values. Maximum exercise capacity and maximum oxygen consumption were normalized within 6 months postoperatively in all groups. There was no difference in recovery of food intake among the groups. Parenteral feeding was associated with the highest rate of nutrition-related complications, whereas enteral feeding reduced quality of life most extensively. CONCLUSION: After major surgery, specialized supportive enteral and parenteral nutrition are not superior to oral nutrition only when guided by a dietitian. PMID- 15880317 TI - Is the monocyte chemotactic protein-1 -2518 G allele a risk factor for severe acute pancreatitis? AB - BACKGROUND & AIMS: Acute pancreatitis (AP) reflects the intensity of the inflammatory response and is divided into mild AP (MAP) or severe AP (SAP). Monocyte chemotactic protein-1 (MCP-1) gene expression is altered by an A/G polymorphism (-2518), with the G allele increasing MCP-1 production. Our aim was to determine whether the MCP-1 -2518 A/G polymorphism affects the severity of AP. METHODS: Seventy-seven consecutive patients and 116 controls were evaluated. The A/G genotype was evaluated by polymerase chain reaction amplification, restriction fragment length polymorphism, and DNA sequencing. MCP-1 serum levels were quantified using a fluorescence bead-based immunoassay. RESULTS: Sixty-three of 77 patients had MAP (82%) and 14 of 77 had SAP (18%). Patients with SAP had a significantly greater proportion of the G allele (12 of 14; 86%) than did control subjects (50 of 116; 43%) (odds ratio [OR], 7.9; 95% confidence interval [CI], 1.7-37, P < .003) or MAP patients (29 of 63; 46%) (OR, 7.0; 95% CI, 1.5-34; P < .007). Patients with pancreatitis and AA genotype had a low risk for SAP (OR, .13; 95% CI, .01-.61; P < .003). As predicted by the genotype, the serum MCP-1 levels were significantly higher in the SAP patients when compared with the MAP patients ( P = .002) and they also predicted death. CONCLUSIONS: MCP-1 -2518 G allele is a risk factor for severe AP. MCP-1 serum levels, measured early in the course of AP, appear to be an accurate predictor of severity of acute pancreatitis and death. PMID- 15880318 TI - Hepatotoxicity and nelfinavir: a meta-analysis. AB - BACKGROUND & AIMS: The inclusion of protease inhibitors in 3-drug highly active antiretroviral regimens for treating patients who are infected with human immunodeficiency virus-1 has had a significant impact in increasing survival and decreasing morbidity. However, the effectiveness of this class of drugs may be compromised by the occurrence of drug-related hepatotoxicity, which is problematic especially in individuals co-infected with hepatitis viruses. Based on its clinical and pharmacologic profile, especially its unique pattern of resistance, nelfinavir has been used frequently as a first-line protease inhibitor therapy for human immunodeficiency virus-1-infected patients. The aim of this study was to identify the relative potential for developing hepatotoxicity for nelfinavir vs other protease inhibitors. METHODS: An exploratory meta-analysis of liver enzyme level increases was conducted in a combined total of 4268 patients derived from 3 large recently conducted prospective and retrospective clinical trials and a prospective cohort study. RESULTS: The results indicate that among 4 commercially available protease inhibitors and a 2-protease inhibitor combination, nelfinavir and indinavir are associated with the lowest rates of occurrence of severe hepatotoxicity (ie, combined estimates of liver enzyme level increases of 2.9% and 3.1%, respectively). The low rate of occurrence of severe hepatotoxicity for nelfinavir was shown even among patients co-infected with hepatitis viruses. CONCLUSIONS: In conclusion, these data provide support for the conclusion that differences in the potential for hepatotoxicity do exist among the commercially available protease inhibitors. PMID- 15880319 TI - Nonsteroidal anti-inflammatory drugs and hepatic toxicity: a systematic review of randomized controlled trials in arthritis patients. AB - BACKGROUND & AIMS: Nonsteroidal anti-inflammatory drugs (NSAIDs) might cause hepatic side effects, but the frequency of these laboratory and clinical side effects is uncertain. METHODS: Searches of bibliographic databases MEDLINE and EMBASE and of public archives of the Food and Drug Administration were conducted to identify randomized controlled trials of diclofenac, naproxen, ibuprofen, celecoxib, rofecoxib, valdecoxib, or meloxicam in adults with osteoarthritis or rheumatoid arthritis that provided information on aminotransferase elevations >3 x upper limit of normal, liver-related discontinuations, hepatic serious adverse events, liver-related hospitalizations, or liver-related deaths. The proportion of patients with each of the hepatic toxicity outcomes was calculated separately by using sample size weighted pooling for each NSAID. RESULTS: Sixty-seven articles from the bibliographic database and 65 studies from the Food and Drug Administration archives met inclusion criteria. Diclofenac (3.55%; 95% confidence interval [CI], 3.12%-4.03%) and rofecoxib (1.80%; 95% CI, 1.52%-2.13%) had higher rates of aminotransferase >3 x upper limit of normal than placebo (0.29; 95% CI, 0.17-0.51) and the other NSAIDs (all < or = 0.43%). The 95% CIs for liver-related discontinuations of all NSAIDs except diclofenac (2.17%; 95% CI, 1.78%-2.64%) overlapped with placebo. Only 1 liver-related hospitalization (among 37,671 patients) and 1 liver-related death (among 51,942 patients) occurred, with naproxen. CONCLUSIONS: Diclofenac and rofecoxib had higher rates of aminotransferase elevations than placebo and other NSAIDs studied. No NSAID studied had increased rates of liver-related serious adverse events, hospitalizations, or deaths. PMID- 15880320 TI - Interferon-stimulated gene expression in black and white hepatitis C patients during peginterferon alfa-2a combination therapy. AB - BACKGROUND & AIMS: Black American patients are less likely to eradicate hepatitis C virus (HCV) infections during treatment with peginterferon (PEG-IFN) and ribavirin. We hypothesized that racial differences in IFN-stimulated antiviral gene induction during treatment might be responsible. METHODS: We examined myxovirus resistance-A (MxA), RNA-dependent protein kinase (PKR), 2'-5' oligoadenylate synthetase (2,5-OAS), and adenosine deaminase-1 (ADAR1) gene expression in the peripheral blood mononuclear cells (PBMCs) of 31 black and 11 white HCV genotype 1 patients at baseline and at weeks 4-12 during PEG-IFN alfa 2a combination treatment. The primary study end point was the early virologic response (EVR)-either an undetectable serum HCV-RNA level or a > or =2-log decrease in serum HCV-RNA level at week 12 compared with week 0. RESULTS: The EVR rate was 67.7% in blacks and 63.6% in whites. Both blacks and whites experienced a significant (200%-500%) increase in 2,5-OAS, MxA, PKR, and ADAR1 expression at treatment weeks 4-12 compared with baseline (P < .01). However, the relationship between IFN-stimulated gene expression and the EVR differed by race. White responders exhibited higher 2,5-OAS and MxA levels at week 4 than white nonresponders (P < .05). IFN-stimulated gene levels did not correlate with EVR in blacks. Black responders had much lower MxA and PKR levels at week 4 than black nonresponders (P < .05). However, black responders maintained increased 2,5-OAS, MxA, and PKR levels from weeks 4-12, whereas the levels decreased to baseline at weeks 8-12 in black nonresponders. CONCLUSIONS: The mechanisms of resistance to PEG-IFN combination therapy may be different in black and white HCV genotype 1 patients. PMID- 15880321 TI - The investigation of hematuria. AB - Persistent microscopic hematuria is present in about 6% of the population, but probably only a small minority have hematuria that does not originate from the glomerulus. Careful analysis of phase-contrast urine microscopy by a skilled observer is critically important in the investigation of hematuria. In glomerular disease, urine microscopy often is second only to renal biopsy examination in helping make a diagnosis. Glomerular and nonglomerular hematuria are distinguished easily on phase-contrast urine microscopy or by an automated peripheral blood cell counter. However, urine microscopy provides additional information about casts and other features that may enable such disparate diagnoses as Fabry's disease, sickle cell disease, and cystine calculi to be made. Macroscopic nonglomerular hematuria is of particular significance because it is much more likely than microscopic hematuria to be associated with malignancy. Macroscopic hematuria originating from the glomerulus indicates the presence of crescentic disease, which requires urgent assessment by renal biopsy examination. We advocate a renal biopsy examination in any individual with a persisting urinary erythrocyte count greater than 100,000/mL. Thirty percent of patients with isolated microscopic hematuria have mesangial immunoglobulin A glomerulonephritis (IgAN) shown on biopsy examination and 20% to 40% of these patients will progress to renal failure without treatment. PMID- 15880322 TI - The epidemiology of thin basement membrane nephropathy. AB - The prevalence of this basement membrane nephropathy (TBMN) may be approximated from the known frequencies of glomerular hematuria in the population, and from the prevalence of autosomal-recessive Alport syndrome and its known relationship to TBMN. These approaches confirm that TBMN affects more than 1% (but < 10%) of the population, making it the commonest inherited renal disease, and one of the commonest conditions affecting the kidney after infections, hypertension, and stones. TBMN is the most frequent cause of persistent glomerular hematuria. Although we do not advocate mass screening for hematuria to detect TBMN, we strongly support investigating hematuria that is discovered incidentally. Individuals with TBMN and isolated hematuria should be evaluated initially by a nephrologist and subsequently reviewed by their family doctor. Those with proteinuria, hypertension, or renal impairment are at risk for progressive renal impairment and should by examined carefully for features of Alport syndrome or an additional glomerular or tubulointerstitial lesion, undergo a renal biopsy examination, be treated symptomatically, and be monitored by a renal physician. PMID- 15880323 TI - The clinical features of thin basement membrane nephropathy. AB - Thin basement membrane nephropathy (TBMN) is a common, lifelong condition affecting the kidneys that is characterized by microscopic glomerular hematuria, minimal or no proteinuria, and normal renal function. It often is discovered incidentally, and usually has an excellent prognosis. Many cases are familial and show autosomal-dominant inheritance. The defining characteristic is a glomerular basement membrane (GBM) that is thinned to about half its normal thickness on ultrastructural examination of the renal biopsy specimen. However, occasionally patients with TBMN develop marked proteinuria or renal impairment. It is unclear whether individuals with TBMN and impaired renal function represent part of the spectrum of TBMN associated with heterozygous COL4A3 or COL4A4 mutations, or if their disease is caused by mutations of other genes, or whether it is caused by a second coexistent renal lesion or is misdiagnosed Alport syndrome. PMID- 15880324 TI - Hematuria in thin basement membrane nephropathy. AB - Thin basement membrane nephropathy (TBMN) often is diagnosed clinically when there is persistent dysmorphic or glomerular hematuria, but minimal proteinuria, normal kidney function, and no other obvious cause. This study investigated hematuria in patients with TBMN. A total of 112 patients with biopsy examination proven TBMN were studied. All had hematuria at the time of presentation, with a mean urinary red blood cell (RBC) count of 256 +/- 250 x 10 3 /mL. Seventy-five (67%) patients attended for review over a median of 48 months (range, 3-120 mo) and provided a total of 485 urine specimens. Twenty-one patients (28%) had no hematuria by phase-contrast microscopy on at least 1 occasion. These corresponded to 32 urine specimens (7% of total). Of the 21 patients, the most recent urinary RBC counts were within the normal range in 11 (52%), but hematuria had recurred in the other 10 (48%). Hematuria is persistent in most patients with TBMN, but occasionally it resolves or is intermittent. PMID- 15880325 TI - Pathology of thin basement membrane nephropathy. AB - Thin basement membrane nephropathy (TBMN) is a glomerular disorder characterized clinically by isolated hematuria and pathologically by diffuse thinning of the glomerular basement membrane (GBM) on ultrastructural examination. The pathologic diagnosis of TBMN is problematic, in part because of the wide range of GBM thicknesses in the normal population. GBM thickness varies with age, sex, and the different methods of tissue preparation and measurement. In addition, there are no standardized diagnostic criteria defining the degree or extent of GBM thinning required for the diagnosis of TBMN. GBM thinning is often seen in other glomerulopathies, where it may represent an overlap with TBMN or may be secondary to GBM damage and remodeling. Importantly, TBMN must be differentiated from the GBM thinning seen in some renal biopsy specimens from boys and female heterozygotes with X-linked Alport syndrome because of the very different prognoses of these two conditions. PMID- 15880326 TI - The nongenetic diagnosis of thin basement membrane nephropathy. AB - Three disorders, thin basement membrane nephropathy (TBMN), immunoglobulin A nephropathy (IgAN), and Alport syndrome (AS), account for the majority of children and adults with persistent glomerular hematuria. Although there is some clinical overlap between these conditions, they can be distinguished on the basis of family history, extrarenal findings, routine immunofluorescence, and glomerular basement membrane ultrastructure or type IV collagen chain composition. This distinction is important because of the very different prognoses of these conditions. PMID- 15880327 TI - The genetics of thin basement membrane nephropathy. AB - The diagnosis of thin basement membrane nephropathy (TBMN) usually is made on the basis of the clinical features or the glomerular membrane ultrastructural appearance. Only now are we beginning to understand the genetics of TBMN and the role of diagnostic genetic testing. The similarity of clinical and glomerular membrane features first suggested TBMN might represent the carrier state for autosomal-recessive Alport syndrome. This was confirmed subsequently by the demonstration that 40% of families with TBMN have hematuria that segregates with the corresponding locus ( COL4A3/COL4A4 ), and identical mutations occur in both conditions. To date, about 20 COL4A3 and COL4A4 mutations have been shown in TBMN, and these mainly are single nucleotide substitutions that are different in each family. The families in whom hematuria does not appear to segregate with the COL4A3/COL4A4 locus cannot all be explained by de novo mutations, and nonpenetrant or coincidental hematuria. This suggests a further TBMN locus. In patients with persistent hematuria, testing for COL4A3 and COL4A4 mutations to diagnose TBMN is problematic because of the huge size of these genes, their frequent polymorphisms, and the likelihood of a further gene locus. It is far more practicable to perform genetic testing to exclude or confirm X-linked Alport syndrome because this condition is the major differential diagnosis of TBMN and has a very different prognosis. PMID- 15880328 TI - The risks of thin basement membrane nephropathy. AB - Most individuals with thin basement membrane nephropathy (TBMN) have an excellent prognosis. For these patients, the only hazards are the anxiety related to misconceptions about the diagnosis and the inconvenience, expense, and wastefulness of unnecessary investigations. However, there also are specific genetic implications for individuals with TBMN because, on average, half their offspring inherit the causative mutations and most of these have hematuria. In addition, despite the generally excellent outcome, some individuals with TBMN develop hypertension, proteinuria, or renal impairment. In some cases, renal failure is caused by apparently progressive but otherwise uncomplicated TBMN, and in others it results from a secondary or coincidental glomerular or tubulointerstitial renal lesion. In particular, TBMN appears to predispose to immunoglobulin (Ig)A glomerulonephritis, and the outcome for these patients is worse than for those with TBMN alone. The risks for patients with TBMN in relation to pregnancy and transplantation have not been well-studied but are described elsewhere in this issue. PMID- 15880329 TI - Thin basement membrane nephropathy associated with other glomerular diseases. AB - Many reports confirm that thin basement membrane nephropathy (TBMN) commonly occurs together with other glomerular diseases such as minimal change glomerulonephritis, diabetes, membranous nephropathy, immunoglobulin (Ig)A glomerulonephritis, and focal segmental glomerulosclerosis. We postulate 3 explanations for these observations. The association of minimal change glomerulonephritis with TBMN probably is artifactual whereas the association with diabetes and membranous glomerulonephritis probably is coincidental. However, the link between TBMN and IgA disease and focal segmental glomerulosclerosis may be pathogenetic. Clinical evidence indicates that the presence of an associated glomerulopathy significantly worsens the prognosis of TBMN. Thus, patients with TBMN and another glomerular lesion usually have more marked proteinuria, and are more likely to have hypertension and renal insufficiency. The frequency of another glomerulopathy in patients with TBMN means that all patients in whom TBMN is suspected but who have heavy proteinuria or renal insufficiency should undergo a renal biopsy examination. However, there is no evidence that TBMN alters the prognosis of another glomerulopathy, and, in particular, patients with TBMN and IgA disease do not have different clinical features or a worse prognosis than those with IgA disease alone. PMID- 15880330 TI - Thin basement membrane nephropathy in pregnancy. AB - There are several published series of pregnancy in patients with nonimmunoglobulin A mesangial proliferative glomerulonephritis (most of whom have thin basement membrane nephropathy [TBMN]). The aim of the present study was to review the maternal and fetal outcomes of pregnancy in women with TBMN. The medical and obstetric histories of 86 women with TBMN and their 182 pregnancies (one twin) were reviewed. Data were collected retrospectively in 164 pregnancies (90%) and prospectively in 18 pregnancies (10%). Hypertension (alone or with proteinuria) developed in 15 unmonitored pregnancies (9%), and proteinuria alone developed in the third trimester in 2 pregnancies (1%). Hypertension was more common in the prospectively monitored pregnancies (6 pregnancies, 33%). In all, there were 174 live births (95%), and all fetal deaths occurred in the first and second trimesters in the absence of maternal complications. However, all the mothers of the 4 small for gestational age babies had been hypertensive. In TBMN, maternal hypertension, prematurity, and small for gestational age rates did not exceed those in the normal population. Overall, pregnancy in women with TBMN does not appear to be attended by a significantly increased maternal or fetal risk. PMID- 15880331 TI - Thin basement membrane nephropathy and renal transplantation. AB - The clinical implications of thin basement membrane nephropathy (TBMN) in renal transplantation must be considered from the perspectives of both the allograft recipient and the donor. Most individuals with TBMN have a benign course, but some develop end-stage renal failure (ESRF) and undergo transplantation. ESRF in patients with TBMN often results from the presence of additional glomerular or interstitial lesions and some of these, such as immunoglobulin (Ig)A disease, may recur in the renal allograft and affect outcome. In addition, individuals with TBMN always must be distinguished from those with glomerular membrane thinning due to Alport syndrome. This is not only to enable appropriate genetic counseling but also to anticipate the possible complication of posttransplant anti glomerular basement membrane disease. From the perspective of the live renal donor, donation from individuals with TBMN (or carriers of X-linked Alport syndrome with thinned membranes) remains controversial because the risks remain unknown. Any effects of the thinned membranes themselves on allograft function are unclear. Further advances in our understanding of the clinical, pathologic, and molecular features of TBMN should result in improved assessment of potential live donors and help stratify those at risk for renal impairment. PMID- 15880332 TI - Bloodletting as a cure for dropsy: heart failure down the ages. AB - BACKGROUND: Dropsy was a term used to describe generalized swelling and was synonymous with heart failure. Its treatment options were scanty and were aimed to cause "emptying of the system" or to relieve fluid retention. These remedies were rudimentary, erratic in action, and associated with inconvenient side effects. METHODS AND RESULTS: Bloodletting, either by venesection or by leeches, was a popular way to alleviate symptoms from dropsy. Although bloodletting, purgatives, cauterization, and Southey tubes were drastic, their use demonstrated that physicians were not powerless to help people with severe heart failure. Several centuries of intensive investigations in different areas of heart failure ended with the development of new therapeutic strategies that made bloodletting obsolete. CONCLUSION: In an era when adequate treatment of heart failure has become a reality, it is appropriate to acknowledge those who paved the way to such great progress. PMID- 15880333 TI - Effect of valsartan on quality of life when added to usual therapy for heart failure: results from the Valsartan Heart Failure Trial. AB - BACKGROUND: The effect on quality of life (QOL) of valsartan administered in addition to prescribed background heart failure therapy was assessed as a secondary endpoint in the Valsartan Heart Failure Trial (Val-HeFT). METHODS AND RESULTS: QOL was assessed in 3010 patients receiving either valsartan (160 mg twice daily) or placebo in addition to prescribed background therapy (beta blockers or angiotensin-converting enzyme inhibitors), using the Minnesota Living with Heart Failure (MLWHF) questionnaire. Treatment differences were compared at intervals to 36 months after randomization and at endpoint (last observation) using analysis of covariance and repeated measures mixed-effects, and at endpoint using a Mantel-Haenszel chi-squared test. Scores lower than baseline were indicative of improved QOL. Valsartan had a significant beneficial effect on the least-square mean change in overall MLWHF score from baseline to study endpoint (+/- standard error) (average followup 23.0 months) compared with placebo (0.19 +/- 0.47 versus 1.94 +/- 0.48; P = .005 respectively). The placebo group was characterized by a deterioration in QOL as the trial progressed. More patients on valsartan reported a clinically meaningful improvement in MLWHF score (a decrease of > or =5 points) than on placebo (34.0% versus 30.2%). CONCLUSION: Valsartan compared to placebo added to prescribed therapy slows progressive worsening of QOL in patients with heart failure. PMID- 15880334 TI - Rationale and design of the multicenter, randomized, double-blind, placebo controlled study to evaluate the Efficacy of Vasopressin antagonism in Heart Failure: Outcome Study with Tolvaptan (EVEREST). AB - BACKGROUND: Hospitalizations for worsening heart failure due to fluid overload (congestion) are common. Agents that treat congestion without causing electrolyte abnormalities or worsening renal function are needed. Tolvaptan is an oral vasopressin (V 2 ) antagonist that decreases body weight and increases urine volume without inducing renal dysfunction or hypokalemia. The Efficacy of Vasopressin antagonism in Heart Failure Outcome Study with Tolvaptan (EVEREST) trial is evaluating mortality, morbidity, and patient-assessed global clinical status in patients treated with tolvaptan compared with standard care. METHODS AND RESULTS: Patients are eligible for inclusion if they have a reduced left ventricular ejection fraction and are hospitalized for worsening heart failure with evidence of systemic congestion. Patients are randomized 1:1 to tolvaptan 30 mg/day or matching placebo for a minimum of 60 days. Time to all-cause mortality and time to cardiovascular mortality or heart failure hospitalization are the coprimary end points. Patient-assessed global clinical status and quality of life are also evaluated. EVEREST will be continued until 1065 deaths occur. As of April 18, 2005, 2260 patients have been enrolled. CONCLUSION: Tolvaptan has been shown to reduce body weight in patients with worsening heart failure without inducing renal dysfunction or causing hypokalemia. The results of EVEREST will determine whether these effects translate into improved clinical outcomes. PMID- 15880335 TI - Temporal trends in survival and hospitalizations in outpatients with chronic systolic heart failure in 1995 and 1999. AB - Abstract Background Community and hospital studies have suggested that survival of patients with heart failure (HF) has increased; however, the causes of the improvement and the hospital readmission rates remain undetermined. Methods and Results We compared survival and hospital admission rates in 2 cohorts enrolled in a national registry of outpatients with HF secondary to left ventricular (LV) systolic dysfunction referred to cardiology centers in 1995 (n = 712) and 1999 (n = 603). One year after enrollment, 163 of 1315 patients (12%) were dead. Survival rates were 85% in the 1995 versus 91% in the 1999 cohort. Older age, New York Heart Association (NYHA) class III-IV, anemia, hyponatremia, hypotension, and a lower LV ejection fraction (LVEF) were associated to an increased risk of all cause mortality by multivariate analysis. Furthermore a significant independent cohort effect was observed: the adjusted risk of death was 1.30 (95% CI 1.16 1.45) for the 1995 versus 1999 cohort (survival difference adjusted P = .0067). The proportion of patients admitted to hospital declined significantly in 1999 versus 1995, for all causes (20% versus 27%, P = .006), for cardiac causes (16% versus 22%, P = .002), and for worsening congestive heart failure (8% versus 15%, P = .0005). Survival free from HF admission was 69% in 1995 versus 84% in 1999 (adjusted P = .0001); NYHA class III-IV, hypotension, diuretics and a lower LVEF were associated to an increased risk of this combined end point by multivariate analysis, as well as the enrollment year (relative risk 1.38, 95% CI 1.22-1.56, P = .0039). Conclusion In a national cardiologic registry of outpatients with systolic HF, survival improved and hospital admissions decreased over a 4-year period. These results underscore the importance of networking and the careful implementation of practice guidelines to elevate standards of care. PMID- 15880336 TI - The VEGF +405 CC promoter polymorphism is associated with an impaired prognosis in patients with chronic heart failure: a MERIT-HF substudy. AB - BACKGROUND: A common polymorphism positioned at +405 of the vascular endothelial growth factor (VEGF) gene is known to influence VEGF protein production. In contrast, a second polymorphism, positioned at -460 polymorphism, has no reported functional effects. VEGF is linked to angiogenesis and might directly influence the clinical outcome of patients with chronic heart failure (CHF). We investigated the association between two VEGF polymorphisms and morbidity and mortality in patients with CHF. METHODS AND RESULTS: VEGF promoter polymorphisms +405 and -460 were examined in 596 CHF patients enrolled in the Metoprolol CR/XL Randomized Intervention Trial in Heart Failure (MERIT-HF) study and in 187 healthy controls. In CHF patients, a risk ratio for each genotype was calculated using the combined endpoint of all-cause mortality or hospitalization. The allele frequencies of the +405 and -460 polymorphisms for the CHF cohort and for 187 healthy controls were not significantly different. However, the presence of the +405 CC genotype (frequency 0.14) was independently associated with an adverse outcome as described by the MERIT study combined endpoint compared with the +405 GG genotype (risk ratio 1.65; 95%CI 1.03-2.64; P = .039). The -460 polymorphism was not associated with an altered prognosis ( P = .60). CONCLUSION: Our results indicate that the VEGF +405 CC genotype is associated with an adverse clinical outcome in patients with CHF. This genotype has been associated with lower plasma VEGF levels, suggesting a possible mechanism of action for the gene variant. This belief is further supported by the fact that the VEGF -460 polymorphism, which does not affect plasma VEGF levels, did not adversely affect the prognosis. PMID- 15880337 TI - Vascular endothelial growth factor and angiogenesis in heart failure. PMID- 15880338 TI - Symptoms in patients with heart failure are prognostic predictors: insights from COMET. AB - BACKGROUND: Although functional status, as assessed by the New York Heart Association classification, is known to be a powerful prognostic marker in chronic heart failure (CHF), the significance of individual symptoms such as breathlessness and fatigue are unknown. OBJECTIVE: To assess the relative importance of self-reported severity of symptoms as predictors of outcomes in CHF. METHODS AND RESULTS: All 3029 patients randomized in the Carvedilol or Metoprolol European Trial (ie, COMET) study were included in the analysis. Mean follow-up was 58 months. Symptoms were assessed by 5-point scales. In a univariate analysis, worse scores for breathlessness, orthopnea and fatigue were all significantly related to increased mortality (all P < .0001) and development of worsening heart failure. In a multivariate Cox regression analysis including 16 baseline covariates, only the symptom of breathlessness remained significantly related to mortality (risk ratio [RR] 1.14 per unit: 95% CI 1.04-1.26; P = .01). Fatigue, but not breathlessness, remained a significant predictor for developing worsening heart failure (RR 1.09 per unit; 95% CI 1.02-1.18; P = .02). CONCLUSIONS: Fatigue and breathlessness, common symptoms in CHF, have important and independent long-term prognostic implications. Accordingly, symptoms need to be effectively evaluated not only because symptom alleviation is a target for treatment, but also because they guide prognosis in patients with CHF. PMID- 15880339 TI - Prospective evaluation of an outpatient heart failure disease management program designed for primary care: the Oregon model. AB - BACKGROUND: Most heart failure care is provided by primary care providers. Although heart failure disease management programs improve outcomes, most have been hospital-based with little integration with primary care providers. To address this issue, a heart failure clinic disease management model was adapted for use in the primary care setting. METHODS AND RESULTS: A heart failure clinic staffed by 2 internists and their nurses was established in a large primary care practice. Medical care and pharmacotherapy were based on national guidelines. Nurses assisted with disease management. Primary outcomes included quality of life, functional class, and all-cause hospital and emergency room admissions 12 months before compared with 12 months after enrollment; a secondary endpoint was patient satisfaction. Of 165 patients sent to the heart failure clinic, 54 were referred back because of no active heart failure, and 18 had only 1 clinic visit. The 93 patients seen 2 or more times had a median age of 75 years. Anti angiotensin II therapy was present in 84% and did not change over time, but doses of angiotensin-converting enzyme inhibitor increased by >50%. beta-blocker use increased from 40% at baseline to 63% at 6 months. Emergency room visits or all cause hospitalizations were reduced (0.86 +/- 1.5 to 0.52 +/- 0.86, P < .001) or trended to be reduced (0.56 +/- 0.98 to 0.35 +/- 0.62, P = .07), respectively, by approximately 40%. Quality of life improved significantly at all time points, and patients were highly satisfied. CONCLUSION: This heart failure disease management model, designed for patients and providers in an primary care setting, was feasible and successful. PMID- 15880340 TI - Assessment of left ventricular systolic function by the chest x-ray: comparison with radionuclide ventriculography. AB - BACKGROUND: The value of the plain chest roentgenogram in predicting cardiac status remains controversial. METHODS AND RESULTS: A total of 111 randomly selected survivors of acute myocardial infarction (age 38 to 83 years) were studied prospectively. X-ray and radionuclide examinations were performed on a morning in the second week after myocardial infarction. From the chest x-ray, left ventricular chamber size and pulmonary vascular congestion were graded visually, and relative cardiac volume was measured to allow for comparison with radionuclide left ventricular end-diastolic volume index (LVEDVI) and left ventricular ejection fraction (LVEF) determined by gated blood pool imaging. Despite significant tendencies for larger radionuclide LVEDVI and lower LVEF with greater radiographic left ventricular size, larger relative cardiac volume, and increasing degrees of pulmonary congestion, wide scatter, and large overlaps between groups precluded reliable radiographic prediction of radionuclide findings. The positive and negative predictive values for radiographic detection of an enlarged LVEDVI ranged from 59% to 80% and 56% to 71%, respectively, and for prediction of a decreased LVEF from 75% to 90% and 40% to 58%, respectively. Accuracy never exceeded 70%. CONCLUSION: Our findings question the value of the chest roentgenogram in the detection and grading of left ventricular systolic dysfunction in patients with recent myocardial infarction. PMID- 15880341 TI - Heart failure in the adult patient with congenital heart disease. AB - BACKGROUND: Adult survival is now the rule for most congenital heart defects. However, despite surgical "repair," heart failure remains a common long-term complication of congenital heart disease. Heart failure specialists are seeing an increasing number of patients with congenital defects as the underlying etiology of their heart failure. METHODS AND RESULTS: Familiarity with the anatomy, physiology, and long-term sequelae of surgical correction are essential for providing care to this complex group of patients. CONCLUSION: This review discusses the diagnosis, pathophysiology, and management of the adult congenital heart patient with concomitant heart failure. PMID- 15880342 TI - Coronary blood flow responses are impaired independent of NO and endothelial function in conscious dogs with dilated cardiomyopathy. AB - BACKGROUND: Dilated cardiomyopathy (DCM) is characterized by nitric oxide (NO) deficiency and endothelial dysfunction. Whether endothelium-independent vasodilation is preserved, particularly in the coronary circulation, remains controversial. METHODS AND RESULTS: We studied systemic and coronary flow responses to the endothelium-dependent agonist, acetylcholine, the cGMP-dependent NO-donor, nitroglycerin, the predominantly endothelium-independent agonist, adenosine, the beta-adrenergic cAMP-dependent agonist, isoproterenol, and the calcium channel antagonist, nicardipine, in conscious dogs with pacing-induced DCM. Systemic blood flow response was impaired to acetylcholine but preserved to other vasodilators in DCM. In contrast, coronary blood flow response was significantly ( P < .05) depressed to all agonists. (Peak coronary blood flow response, control versus DCM: acetylcholine: 221 +/- 14% versus 156 +/- 11%; nitroglycerin: 220 +/- 17% versus 138 +/- 9%; adenosine: 635 +/- 65% versus 376 +/- 56%; nicardipine: 338 +/- 59% versus 115 +/- 23%; isoproterenol: 219 +/- 18% versus 86 +/- 20%). The attenuation was independent of systemic hemodynamic differences. CONCLUSION: In contrast to systemic responses, coronary blood flow responses in DCM are impaired dependent or independent of NO or second messenger mechanisms, implying either distal signaling defects or structural abnormalities in the coronary vasculature. PMID- 15880344 TI - Elevated CXCL16 expression by synovial macrophages recruits memory T cells into rheumatoid joints. AB - OBJECTIVE: Directional migration of leukocytes is orchestrated by the regulated expression of chemokine receptors and their ligands. The receptor CXCR6 is abundantly expressed by Th1-polarized effector/memory lymphocytes accumulating at inflammatory sites. This study was undertaken to examine the presence of CXCR6+ T cells and of CXCL16, the only ligand for CXCR6, in the joints of patients with rheumatoid arthritis (RA). METHODS: Flow cytometry analysis of the expression of CXCR6 by peripheral blood and synovial fluid (SF) T cells. In addition, by performing conventional and real-time reverse transcriptase-polymerase chain reaction, immunohistochemistry, and enzyme-linked immunosorbent assay, we determined the expression of CXCL16 and its protease ADAM-10 within synovium and by cultured macrophages. SF T cell migration was studied with the Transwell system. RESULTS: Accumulation of CXCR6+ T cells within RA SF coincided with highly elevated levels of CXCL16+ macrophages. In vitro studies revealed that monocytes started to express CXCL16 upon differentiation into macrophages, and that RA SF and tumor necrosis factor (TNF) enhanced CXCL16 expression. Moreover, RA patients responding to anti-TNF therapy showed a strongly decreased CXCL16 expression, whereas nonresponding patients did not. Interestingly, ADAM-10, a recently identified protease of CXCL16, was abundantly expressed by CXCL16+ macrophages in vitro and in RA in vivo, which resulted in increased levels of cleaved CXCL16 in RA SF relative to controls. Finally, CXCR6+ T cells from RA SF were attracted by CXCL16. CONCLUSION: These data provide evidence that enhanced production of CXCL16 in RA synovia leads to recruitment of CXCR6+ memory T cells, thereby contributing to the inflammatory cascade associated with RA pathology. PMID- 15880345 TI - Kappa and delta opioid receptors are expressed but down-regulated in fibroblast like synoviocytes of patients with rheumatoid arthritis and osteoarthritis. AB - OBJECTIVE: To investigate the expression and regulation of the kappa-opioid receptor (KOR) and the delta-opioid receptor (DOR) in fibroblast-like synoviocytes (FLS) from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), and to explore the potential antiarthritic mechanisms of peripheral KORs. METHODS: FLS isolated from synovial tissues of 6 OA patients, 8 RA patients, and 2 healthy individuals were exposed to the selective KOR agonist U69593, the selective DOR agonist SNC 80, and kappa-opioid dynorphin A in the presence or absence of the KOR antagonist nor-binaltorphimine, the DOR antagonist naltrindole, and the proinflammatory cytokines tumor necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta). The expression of KOR and DOR in OA and RA FLS was evaluated on the messenger RNA (mRNA) and protein levels with TaqMan real-time reverse transcriptase-polymerase chain reaction and immunofluorescence staining, respectively. KOR/DOR-mediated activation of ERK-1 and ERK-2 was investigated by Western blotting. RESULTS: We detected functional KOR and DOR in normal FLS and observed a reduction of both receptors in OA and RA FLS, which was more distinct in RA FLS. U69593 enhanced KOR mRNA expression in both OA and RA FLS in a KOR antagonist-reversible manner. However, the dose required for maximal enhancement in RA FLS was 10 times higher than that required in OA FLS. TNFalpha and IL-1beta both suppressed the expression of DOR and KOR mRNA in both OA and RA FLS. CONCLUSION: DOR and KOR are constitutively present in normal FLS and are suppressed under inflammatory conditions, such as RA and OA. Most interestingly, the KOR agonist U69593 may exert an antiarthritic effect via up-regulation of KOR in OA and RA FLS. PMID- 15880346 TI - Knee height, knee pain, and knee osteoarthritis: the Beijing Osteoarthritis Study. AB - OBJECTIVE: Few risk factors for knee osteoarthritis (OA) are appreciated, and the discordance between symptoms and the severity of structural disease has not been explained. Knee height contributes to moments around the knee. The longer the leg, the more torque is present. Although this would suggest that having long legs would be related to the occurrence of knee OA and pain, this issue has not been studied. Our aim was to explore the association between knee height, knee pain, and knee OA. METHODS: We recruited a random sample of Beijing residents ages 60 years and older. Subjects answered questions about joint symptoms, and radiographs of their knees were obtained. A knee joint with a Kellgren/Lawrence grade of >/=2 was defined as having radiographic OA. Patellofemoral OA was defined as being present when grade >/=1 osteophytes or grade >/=1 joint space narrowing was observed on skyline views of the patella or anterior femur. Subjects were considered to have symptomatic OA when both radiographic OA and self-reported pain were present in the same joint. Knee height was measured on the right leg using a sliding broad-blade caliper; the subject was seated, and the subject's feet were bare. We used logistic regression analyses to assess whether knee height was associated with prevalent radiographic and symptomatic OA. We then assessed whether knee height was associated with knee symptoms independently of structural change. RESULTS: A total of 1,006 men (mean +/- SD age 68.4 +/- 6.4 years) and 1,500 women (mean +/- SD age 67.5 +/- 6.1 years) participated in this study. Higher knee height was associated with an increasing prevalence of both radiographic and symptomatic OA, especially among women. For radiographic OA, the magnitude of association was similar for the patellofemoral and tibiofemoral compartments. Among women with knee pain, higher knee height was associated with more severe knee pain (P = 0.0004 for the highest quartile versus the lowest quartile of knee height) independently of the severity of radiographic OA. CONCLUSION: Knee height is associated with prevalent radiographic and symptomatic knee OA. It may also play an important role in knee symptoms. This study highlights the importance of mechanical forces in the determination of OA and knee symptoms. PMID- 15880347 TI - Relationship of radiographic and clinical variables to pinch and grip strength among individuals with osteoarthritis. AB - OBJECTIVE: Little is known about how specific radiographic features are related to hand strength in osteoarthritis (OA). This study examined associations of radiographic variables with pinch and grip strength among individuals with radiographic hand OA. METHODS: Participants (n = 700, 80% female, mean age 69 years) were part of a study on the genetics of generalized OA. All had bilateral radiographic hand OA. Linear models were used to examine associations of grip and pinch strength with 1) OA in joint groups (proximal interphalangeal, metacarpophalangeal [MCP], carpometacarpal [CMC]), 2) OA in rays (first through fifth), and 3) summed Kellgren/Lawrence (K/L) grades for severity of OA in all joints. Adjusted models controlled for age, sex, hand pain, chondrocalcinosis, and hand hypermobility. Mixed models accounted for clustering within families. RESULTS: In bivariate analyses, all joint groups, all rays, and total summed K/L grades were significantly negatively associated with grip and pinch strength (P < 0.05). In adjusted models, the only joint group significantly associated with grip strength was the CMCs, and only OA in the MCP joint was significantly associated with pinch strength (P < 0.05). The only ray significantly associated with grip strength (P < 0.05) was ray 1, and no individual rays were significantly associated with pinch strength. A higher summed K/L grade was significantly associated with both lower grip strength and lower pinch strength. CONCLUSION: Among individuals with radiographic hand OA, increasing radiographic severity is associated with reduced grip and pinch strength, even when controlling for self-reported pain. Individuals with radiographic OA in specific locations (CMC joints, MCP joints, and ray 1) may be at particular risk for reduced hand strength. PMID- 15880348 TI - ADAMTS-1-knockout mice do not exhibit abnormalities in aggrecan turnover in vitro or in vivo. AB - OBJECTIVE: To determine the role of the proteinase ADAMTS-1 in normal and accelerated catabolism of aggrecan in articular and growth plate cartilage of mice. METHODS: Expression of ADAMTS-1 was determined using reverse transcriptase polymerase chain reaction (RT-PCR) analysis of RNA isolated from microdissected chondrocytes from different zones of mouse growth plate and articular cartilage. Real-time RT-PCR for ADAMTS-4, ADAMTS-5, and ADAMTS-9 was performed on femoral head cartilage of wild-type (WT) and ADAMTS-1-knockout (KO) mice. Histologic and immunohistologic evaluation of growth plate and articular cartilage was performed in WT and KO mice from birth to 12 weeks of age. The effect of ADAMTS-1 ablation on cartilage proteoglycan loss was studied in antigen-induced arthritis (AIA). Aggrecan catabolism in WT and KO mice was studied in an in vitro model of cartilage degradation, by quantitation of glycosaminoglycan loss and histologic, immunohistologic, and Western immunoblot analyses. RESULTS: ADAMTS-1 messenger RNA (mRNA) was expressed in normal mouse articular and growth plate cartilage and was up-regulated in terminal hypertrophic differentiation of growth plate chondrocytes. There was no difference in mRNA levels in the cartilage of WT compared with KO mice for the other potential aggrecanases ADAMTS-4, ADAMTS-5, or ADAMTS-9. ADAMTS-1-KO mice were significantly smaller than their WT littermates; however, no morphologic differences between the genotypes were evident in growth plate or articular cartilage from birth to skeletal maturity (12-16 weeks). Similarly, no difference in cartilage aggrecan content or presence of aggrecan degradation products was detected between WT and KO mice. There was no difference between WT and KO mice in the degree of synovial inflammation or depletion of cartilage aggrecan in AIA. There was no difference between WT and KO cartilage in either basal or stimulated aggrecan loss in vitro; however, subtle changes in the aggrecanase-generated aggrecan catabolites were observed in interleukin-1-treated cartilage. CONCLUSION: Although ADAMTS-1 is expressed in articular and growth plate cartilage and is able to cleave aggrecan at physiologically relevant sites, our results indicate that it does not play a significant nonredundant role in normal cartilage and bone development and growth. Similarly, ablation of ADAMTS-1 offered no protection from accelerated aggrecanolysis in an inflammatory model of arthritis or in an in vitro model of early cartilage degradation. ADAMTS-1 does not appear to be a viable target for treatment of cartilage destruction in arthritis. PMID- 15880349 TI - Type I collagen alpha1 Sp1 transcription factor binding site polymorphism is associated with reduced risk of hip osteoarthritis defined by severe joint space narrowing in elderly women. AB - OBJECTIVE: A common G/T substitution at an Sp1 binding site in intron 1 of the COL1A1 gene has been reported to be associated with reduced bone mineral density and increased risk of osteoporotic fracture. The purpose of this study was to examine whether there is an association between COL1A1 Sp1 polymorphism and radiographic osteoarthritis (OA) of the hip in elderly women in the Study of Osteoporotic Fractures. METHODS: Radiographic hip OA status of subjects was defined by the presence of 1 of the following criteria in either hip: a joint space narrowing (JSN) score of >/=3, a Croft summary grade of >/=3, or both definite (score >/=2) osteophytes and JSN in the same hip. Cases of radiographic OA of the hip were further subdivided into those with JSN score >/=3 and those with a femoral osteophyte score >/=2 and JSN score /=3), and 131 (23%) had moderate or moderate-to severe femoral osteophytosis (score >/=2). There was no association of the T/T genotype with either radiographic hip OA or radiographic hip OA characterized by osteophytosis. For radiographic OA of the hip characterized by moderate-to-severe JSN, the odds of disease were significantly reduced among subjects with the T/T compared with the G/G genotype (OR 0.30, 95% CI 0.11-0.81, P = 0.02) and did not change after adjustment for potential confounders (OR 0.36, 95% CI 0.13-0.99, P = 0.048). CONCLUSION: The T/T genotype of the COL1A1 Sp1 polymorphism was associated with a reduced risk of radiographic OA of the hip characterized by JSN. This association should be confirmed in other populations to determine if mechanistic studies are warranted. PMID- 15880350 TI - Incidence of vasculitis in rheumatoid arthritis: comment on the article by Turesson et al. PMID- 15880351 TI - SCN1A mutations and epilepsy. AB - SCN1A is part of the SCN1A-SCN2A-SCN3A gene cluster on chromosome 2q24 that encodes for alpha pore forming subunits of sodium channels. The 26 exons of SCN1A are spread over 100 kb of genomic DNA. Genetic defects in the coding sequence lead to generalized epilepsy with febrile seizures plus (GEFS+) and a range of childhood epileptic encephalopathies of varied severity (e.g., SMEI). All published mutations are collated. More than 100 novel mutations are spread throughout the gene with the more debilitating usually de novo. Some clustering of mutations is observed in the C-terminus and the loops between segments 5 and 6 of the first three domains of the protein. Functional studies so far show no consistent relationship between changes to channel properties and clinical phenotype. Of all the known epilepsy genes SCN1A is currently the most clinically relevant, with the largest number of epilepsy related mutations so far characterized. PMID- 15880352 TI - Prevalence estimates of recurrent balanced cytogenetic aberrations and gene fusions in unselected patients with neoplastic disorders. AB - Chromosome abnormalities have been reported in more than 46,000 benign and malignant neoplastic disorders, leading to the identification of numerous recurrent abnormalities. A substantial number of recurrent balanced aberrations (RBAs), in particular, reciprocal translocations, occur with remarkable specificity in association with clinical and tumor characteristics. This information has become increasingly important both in basic cancer research, as a means to identify pathogenetically important genes, and clinically, as a diagnostic and prognostic instrument. Knowledge of the frequencies of such aberrations thus is of theoretical as well as practical value. However, it is unknown to what extent the data available in the literature reflect reality. A large proportion of the published cases, at least 40%, are biased, in the sense that they were reported because of a specific or unusual karyotypic feature. We have systematically ascertained all RBAs and present data on the frequencies of these abnormalities and their molecular genetic consequences among unselected patients, that is, those studied as part of investigations of consecutive series of individuals with a particular neoplastic disorder. The salient features of the present study are: (1) published data clearly overestimate the prevalence of individual RBAs in most tumor types as well as the proportion of patients having such aberrations. In fact, several well-known published RBAs are not recurrent or have not even been seen among unselected patients, and in no tumor entity, except for chronic myeloid leukemia, does the frequency of unselected cytogenetically abnormal neoplasms with RBAs exceed 35%; (2) the proportions of unselected cases characterized by RBAs among those tumor entities in which at least one RBA has been identified vary considerably both within and among hematologic malignancies, malignant lymphomas, and solid tumors; and (3) the molecular consequences of a substantial proportion, ranging from 19% in hematologic malignancies to 65% in epithelial tumors, of the most common RBAs in unselected patients remain to be clarified. PMID- 15880353 TI - Short-term interleukin-1(beta) increases the release of secreted APP(alpha) via MEK1/2-dependent and JNK-dependent alpha-secretase cleavage in neuroglioma U251 cells. AB - Several lines of neuroimmunological evidence correlate the development of the inflammatory responses of the brain with the formation of amyloid plaques associated with the pathogenesis of neurodegenerative disorders such as Alzheimer's disease. Within this context, we tested the ability of interleukin 1beta (IL-1beta) to regulate the processing of beta-amyloid precursor protein (beta-APP) in neuroglioma U251 cells. Our findings have shown that short-term treatment with IL-1beta (2 hr) resulted in a concentration-dependent decrease in the amount of the cell-associated form of beta-APP in U251 cells as compared to untreated cells, whereas a 2-hr treatment with IL-1beta led to increased release of secreted APP(alpha) fragment (sAPP(alpha)) into the conditioned media of the cells. The fact that sAPP(alpha) is an alpha-secretase cleavage metabolite of the cell-associated form of beta-APP, and the observation that IL-1beta-induced sAPP(alpha) release could be blocked by tissue inhibitors of metalloproteinases-1 (alpha-secretase inhibitors), suggested that alpha-secretase might be involved in IL-1beta-induced-sAPP(alpha) release. Moreover, to determine whether an intracellular signaling pathway mediates the IL-1beta-induced increase in sAPP(alpha) secretion, we used various specific signaling inhibitors and found that sAPP(alpha) release is significantly blocked by the mitogen-activated protein kinase (MEK1/2) inhibitor PD98059 and the c-Jun N-terminal kinase inhibitor SP600125. These findings suggested that the mechanism of IL-1beta induced-sAPP(alpha) release is dependent on MEK1/2- and JNK-activated alpha secretase cleavage in neuroglioma U251 cells. PMID- 15880354 TI - The Italian Registry for Therapeutic Apheresis. A report from the Apheresis Study Group of the Italian Society of Nephrology. AB - Many clinical indications and different technical issues have been reported on therapeutic apheresis: much criticism has also been recorded in several instances, mainly due to the lack of large clinical trials to validate collected data. A Registry where all the available data can be organized and analyzed therefore becomes a priority for all the professionals involved in apheresis. The purpose of this report is to describe the data submitted from 1994 to 2004 from 15,285 treatments on 1,477 patients from 44 Centers, including mainly, but not exclusively, Nephrological Units, collected by the Apheresis Study Group of the Italian Society of Nephrology in 15 Italian regions. Plasma exchange accounted for 56.2% of the procedures, and of these 50.4% were performed by filtration. Plasma treatment was used in 40.1% of procedures, namely with Protein A immunoadsorption (14.6%), LDL-Cholesterol dextran sulfate adsorption (9.7%), and semiselective cascade or double filtration (12.6%). Cell apheresis, limited to photopheresis, was used in 0.85% of cases, and whole blood treatment (direct adsorption lipoprotein, and molecular adsorption recirculating system) in 2.7%. The procedures analyzed here account for less than 20% of estimated therapeutic apheresis performed in Italy, according to the national survey of activity performed for year 2000 by the Italian Apheresis Society. Notwithstanding that the data are largely incomplete, they are sufficiently informative for a definite trend: plasma treatment with filtration on fractionation filters and adsorption must be used as often as possible, instead of plasma exchange, thus obtaining the most selective removals. PMID- 15880355 TI - Complications of donor apheresis. AB - A decreasing blood donor pool in the presence of increasing blood transfusion demands has resulted in the need to maximally utilize each blood donor. This has led to a trend in the increasing use of automated blood collections. While apheresis donation shares many reactions and injuries with whole blood donation, because of the differences, unique complications also exist. Overall, evidence in the literature suggests that the frequency of reactions to apheresis donation is less than that seen in whole blood donation, though the risk of reactions requiring hospitalization is substantially greater. The most common apheresis specific reaction is hypocalcemia due to citrate anticoagulation, which, while usually mild, has the potential for severely injuring the donor. Other reactions to apheresis donation are uncommon (e.g., hypotension) or rare (e.g., air embolism). More worrisome, and in need of additional study, are the long-term effects of apheresis donation. Recent evidence suggests that repeated apheresis platelet donations may adversely effect thrombopoiesis as well as bone mineralization. Granulocyte donation has also been implicated in unexpected long term consequences. PMID- 15880356 TI - Donor leukocyte infusions and graft-versus-malignancy. PMID- 15880357 TI - Second primary tumors in children and young adults in the North of England (1968 99). AB - BACKGROUND: This study describes the risk of second malignancy in patients diagnosed with cancer under the age of 25 years, registered on the Northern Region Young Person's Malignant Disease Registry. PROCEDURE: Incidence rates were calculated to describe the occurrence of second malignancies, rate ratios were estimated to compare rates between subgroups. Standardized incidence ratios (SIRs) were calculated for comparison with a reference population. RESULTS: There were 4,072 children and young adults diagnosed with a first malignancy from 1968 to 1999, of whom 68 had a second malignancy (including basal cell carcinomas and meningiomas). The incidence rate of second malignancy is 1.7 per 1,000 survivor person-years (95% CI: 1.4, 2.2), reflecting a four-fold increased risk of malignancy compared with the general population. The rate of second malignancy was non-significantly higher for those diagnosed during young adulthood rather than childhood (RR = 1.2, 95% CI: 0.7, 2.0), significantly higher in females than males (RR = 1.8, 95% CI: 1.1, 3.0) and significantly lower for those diagnosed in more recent years (RR = 0.4, 95% CI: 0.2, 0.8). In contrast, the SIRs indicated that children were at substantial increased risk; whilst males and females, and those diagnosed in earlier and later time periods, were at equivalent risks. CONCLUSIONS: There is evidence of a sustained increased risk of second malignancy in those treated for primary cancer, especially those diagnosed in childhood; with no evidence that this risk is reducing. PMID- 15880358 TI - Butyrate response factor 1 enhances cisplatin sensitivity in human head and neck squamous cell carcinoma cell lines. AB - Cisplatin is a widely used chemotherapeutic agent in head and neck squamous cell carcinoma (HNSCC). Resistance to cisplatin is a common feature of HNSCC. To identify genes that may regulate cisplatin sensitivity, we carried out a cDNA microarray analysis of gene expression in cisplatin-sensitive and cisplatin resistant HNSCC-derived cell lines. Among genes differentially expressed by cisplatin treatment, we have confirmed the elevated expression of butyrate responsive factor 1 (BRF1) in cisplatin-sensitive HNSCC cells and have demonstrated that the expression level of BRF1 is associated with cisplatin sensitivity. Specific inhibition of BRF1 expression using an antisense oligodeoxynucleotide (ODN) decreased the cisplatin-sensitivity and, on the contrary, overexpression of BRF1 increased cisplatin-sensitivity in HNSCC cells. Elevated expression of BRF1 decreased the level of the human inhibitor of apoptosis protein-2 (cIAP2) and increased the caspase-3 activity in HNSCC cells. In addition, elevated expression of BRF1 decreased the expression level of enhanced green fluorescent protein (EGFP) linked to a 3' terminal AU-rich element (ARE) of cIAP2 mRNA. These findings demonstrate that BRF1 expression enhanced cisplatin sensitivity in HNSCC cells by reducing the levels of cIAP2 mRNA. PMID- 15880359 TI - Delivery of therapeutic apheresis services in the United States. PMID- 15880360 TI - World apheresis registry. PMID- 15880361 TI - Apheresis activity in Venezuela. AB - Interest for apheresis activity has been growing in Venezuela. In 1976 there were only a few devices; in 2003, 80 apheresis machines performed 27,675 donor apheresis procedures and 547 therapeutic procedures countrywide. We report the activity at the Metropolitan Blood Bank (the largest one of the country) in the period 1999-2003: 597 therapeutic procedures were performed in 171 patients, during 212 crisis episodes. The average age was 38 +/- 16 years, 65% male and 35% female. Most of the therapeutic procedures were therapeutic plasma exchange for hematology diseases (mainly thrombotic thrombocitopenic purpura and hemophilia inhibitors), including 184 therapeutic procedures with the Autopheresis-C (Baxter Healthcare Corp., Deerfield, IL). Most common adverse effects (3.9%) were hypotension and allergic reactions to the plasma. PMID- 15880362 TI - Cancer risk perceptions in an urban Mediterranean population. AB - The objective of our study was to analyze the perceived (belief) or adopted (behavior) measures to reduce cancer risk in a Spanish population. We used cross sectional data from the Cornella Health Interview Survey Follow-up Study (CHIS.FU). We analyzed 1,438 subjects who in 2002 answered questions about risk perceptions on cancer and related behavior (668 males and 770 females). The benefits of avoiding cigarette smoking (95.8%), sunlight exposure (94.9%) and alcohol (81.0%) were widely recognized. On the other hand, electromagnetic fields (92.1%), food coloring and other food additives (78.4%) or pesticides (69.4%), whose role in cancer occurrence, if any, remain unproven, were clearly considered as cancer risk factors in this population. Compared to men, women more frequently reported healthy behaviors, and the role of exogenous factors (i.e., environmental risk factors) were widely popular. There was a socioeconomic gradient on cancer risk perception with respect to several lifestyle or dietary factors. Individuals with higher educational level scored lower in several risk factors than those with primary or less than primary school education. Smokers reported adopting fewer healthy behaviors than former or never smokers. How people perceive health issues and risk or make choices about their own behavior does not always follow a predictable or rational pattern. PMID- 15880363 TI - Inhibition of MDA-MB-231 cell cycle progression and cell proliferation by C-2 substituted oestradiol mono- and bis-3-O-sulphamates. AB - A natural metabolite of oestradiol (E2), 2-methoxyoestradiol (2-MeOE2), exerts both antitumour and antiangiogenic effects. 2-MeOE2 is currently in clinical trials for the treatment of a variety of cancers. We have previously shown that a number of sulphamoylated analogues of 2-MeOE2 possess enhanced potency and bioavailability with respect to 2-MeOE2. In our study, the effects of C-2 substituted E2 derivatives, with sulphamoylation at the C-3 and/or C-17 position, on ERalpha -ve MDA-MB-231 breast cancer cells were evaluated. Sulphamoylated derivatives were potent inhibitors of cell proliferation, and these effects were irreversible when compared to growth inhibitory effects induced by 2-MeOE2. Cell cycle analysis suggested that these derivatives caused cells to arrest at the G2 M phase of the cell cycle. Sulphamoylated analogues suppressed the clonogenic potential of MDA-MB-231 cells and also their growth on Matrigel culture substratum. Immunofluorescence studies showed fragmented nuclear bodies and an abnormal microtubule cytoskeleton in cells exposed to one of the potent compounds, 2-MeOE2-bis-sulphamate. In addition, these analogues induced phosphorylation of BCL-2, a protein considered to be the guardian of microtubule integrity. In each of the assays, the sulphamoylated derivatives were at least 10 fold more potent than the parent compound 2-MeOE2. In view of the enhanced potencies associated with sulphamoylated E2 derivatives in ERalpha -ve cells, these analogues should hold considerable therapeutic potential for the treatment of hormone-independent breast cancers. PMID- 15880364 TI - Long-term effects of LDL apheresis in patients with severe hypercholesterolemia. AB - Familial hypercholesterolemia (FH) is an inherited disorder of lipoprotein metabolism involving mutations in the LDL receptor (LDL-R). Patients with mutation in one (heterozygous) or both (homozygous) genes have markedly elevated LDL cholesterol and are at increased risk for coronary heart disease (CHD). Aggressive lipid lowering is required for homozygous and many heterozygous FH patients. This often involves LDL-apheresis, where LDL and other apo-B containing lipoproteins are selectively removed from the plasma. We have retrospectively studied 34 patients treated with biweekly LDL-apheresis at the Hospital of the University of Pennsylvania. In our patient population, adverse events were uncommon and rarely resulted in shortened treatment time. There was a dramatic decrease in the relative risk of cardiovascular events and cardiovascular interventions in patients treated with LDL-apheresis for an average of 2.5 years. Some but not all patients had long-term reduction in their LDL levels as a result of LDL-apheresis, suggesting that time-averaged reduction in LDL and/or LDL:HDL ratios were responsible for clinical improvement. These data support the use of LDL-apheresis in patients with FH, as well as medication-intolerant patients that have elevated LDL cholesterol despite maximal pharmacological treatment. PMID- 15880365 TI - Apheresis in the Philippines. AB - This is a retrospective study of 131 patients who underwent 479 therapeutic apheresis (TA) procedures at St. Luke's Medical Center, in the Philippines, from 1994 to 2003. Diseases were classified according to indication categories using the American Society for Apheresis (ASFA) guidelines. Seventy-eight percent of the patients belonged to ASFA Category I, 14% to Category II, 8% to Category III, and none in Category IV. The most common indication was for neurological disorders (62%), mainly acute inflammatory demyelinating polyneuropathy (49%) and myasthenia gravis (40%). The remaining disease groups were hematologic (25%), renal and metabolic (8%), and autoimmune and rheumatic diseases (5%). Oral corticosteroids are the preference of many for the treatment of CIDP, being much less expensive than IVIG infusion or TA. Favorable response is noted in the use of the latter to prevent kidney transplant rejection through antibody removal among pre-sensitized patients before transplantation. Complete recovery was seen in 2 of 4 patients with acute hepatic failure. The negative outcome of controlled studies in the literature has limited our utilization of TA for the treatment of systemic lupus erythematosus. Immunoadsorption for idiopathic thrombocytopenic purpura has not been performed locally due to the unavailability and prohibitive cost of affinity columns. Factors affecting the practice of TA in our country are cited. Organization of a multicenter study group and an apheresis national registry may lead to a better future of TA in the Philippines. PMID- 15880366 TI - Bacterial detection in platelet components and the rationale for pathogen inactivation: a blood center perspective. PMID- 15880367 TI - Black tea induces tumor cell apoptosis by Bax translocation, loss in mitochondrial transmembrane potential, cytochrome c release and caspase activation. AB - Recently the anti-cancer role of black tea has gained immense importance. Nevertheless, the signaling pathways underlying black tea-induced tumor cell death are still unknown. Previously we reported that black tea induces Ehrlich's ascites carcinoma (EAC) cell apoptosis by changing the balance between pro-and anti-apoptotic proteins. It is now well accepted that many cell death pathways converge at the mitochondria to decrease mitochondrial transmembrane potential (MTP) thereby releasing apoptogenic proteins and resulting in the activation of effecter caspases responsible for the biochemical and morphological alterations associated with apoptosis. The role of pro-apoptotic protein, Bax, in initiating mitochondrial death cascade has also been established. Here we demonstrate that in culture black tea extract induces EAC apoptosis in a dose-dependent manner- with IC50 at 100 microg/ml. At this dose, intracellular Bax level increases in EAC followed by its translocation from cytosol to mitochondria resulting in loss in MTP. A search for the downstream pathway further reveals that black tea induces mitochondrial cytochrome c release and activates caspases 9 and 3 by 2 pathways, a) independent of and b) dependent on MTP loss. Interestingly, Black tea-induced death signal might probably be amplified through mitochondrial membrane depolarization via a feedback activation loop from caspase 3. All these findings indicate that black tea initiates mitochondrial death cascade in EAC cells and thereby results in EAC apoptosis. PMID- 15880368 TI - New stool tests for colorectal cancer screening: a systematic review focusing on performance characteristics and practicalness. AB - New stool tests may be promising tools for future colorectal cancer (CRC) screening. The aim of this review was to summarize current evidence of performance characteristics and practicalness in a population-based screening setting of recently developed stool tests. The MEDLINE database was searched for relevant articles published until July 2004. Studies were included if they comprised more than 10 cases and more than 10 controls. Details on study population, performance characteristics and stool collection procedure were taken into account. Overall, 29 studies, mostly retrospective, were included, investigating 17 different stool markers or marker combinations. Underlying study populations were very heterogeneous and mostly very small. Half of the studies reported sensitivity for adenomas in addition to sensitivity for CRC, and fewer than half reported sensitivity by tumor stage or location. Performance characteristics of stool tests varied to a large extent. For most DNA-based markers, specificity was about 95% or higher, but sensitivity was mostly low even for invasive CRC. More studies with larger sample sizes were done for protein based markers, which typically had lower specificity. In most studies, stool samples were frozen within a rather short time period after defecation. While promising performance characteristics have been reported for some tests, more pervasive evidence from larger, prospectively designed studies, which also consider aspects of practicalness, e.g., the possibility of mailing the samples, is needed. PMID- 15880369 TI - Blood component collection by apheresis. AB - Apheresis component collection is a rapidly growing area in the blood collection field. Several instruments with varying capabilities are available. This is a brief review of the equipment available for granulocyte and apheresis component collection and indications for their use. In the United States, granulocytes are collected with the Fenwal CS3000, Fenwal CS3000 Plus, COBE (Gambro) Spectra, Haemonetics LN9000, and Fresenius AS 104. The use of hetastarch for sedimenting agent and stimulation with G-CSF and G-CSF plus dexamethasone have substantially increased granulocyte yields. Plateletapheresis is performed in the United States on the Fenwal CS3000, Fenwal CS3000 Plus, Fenwal Amicus, COBE (Gambro) Spectra, Gambro Trima Version 4, Gambro Trima Accel (Version 5), and Haemonetics LN9000. Automated red blood cell (RBC) collections are performed with the Haemonetics MCS+LN8150, Gambro Trima Version 4, Gambro Trima Accel (Version 5), Amicus, and Baxter Alyx. The RBC can be collected concurrently (with other components) in some instruments or separately in others. Plasma is collected concurrently on several instruments. Plasmapheresis for plasma only is performed on the Fenwal Autopheresis C and Haemonetics PCS2. Granulocyte yields range from 0.46 x 10(10) to 1.0 x 10(10) for unstimulated donors and 2.1 x 10(10) to 2.6 x 10(10) for donors stimulated with dexamethasone or prednisone. The use of G-CSF and G-CSF with dexamethasone has substantially increased granulocyte yields with yields of 4.1 x 10(10) to 10.8 x 10(10) reported. Platelet collection rates of 0.045-0.115 x 10(11) plt/min have been reported. Collection efficiencies of 46-85.7% have been reported. Automated (apheresis) component collection has the advantages of controlled volumes or doses of component, efficient use of the donor, multiple components from the same donor, better inventory control, and better quality control due to less manipulation of the individual components. Disadvantages of automated component collection include the use of expensive equipment and disposables, the need for specially trained machine operators, and lower capacity to collect large volumes of blood compared to whole blood donation. The use of apheresis component collection is rapidly growing to provide the best blood components in the most efficient manner. PMID- 15880370 TI - Schimke-immuno-osseous dysplasia: new mutation with weak genotype-phenotype correlation in siblings. AB - Schimke-immuno-osseous dysplasia (SIOD) is a multisystem disorder with the following consistent clinical features: spondyloepiphyseal dysplasia with disproportional growth deficiency, nephrotic syndrome with focal and segmental glomerulosclerosis, and defective cellular immunity. Transitory ischemic attacks due to vaso-occlusive processes are complications in some patients with severe SIOD. Recently, mutations of SMARCAL1, which encodes a putative chromatin remodelling protein, have been associated with SIOD. Patients with milder disease were observed to harbor a missense mutation on each allele, whereas patients with a severe form of the disease were predicted to have at least one allele with a nonsense, frameshift or splicing mutation. We report two brothers who are both compound heterozygous for the mutations 836 T>C and 2542 G>T detected in exons 4 and 17, respectively. We demonstrate the lack of genotype-phenotype correlation in these patients as one brother shows some features of the severe form while the other does not. Neither clinical nor molecular findings can fully predict the clinical course of SIOD. PMID- 15880371 TI - Evaluation of fungicides for the control of carrot cavity spot. AB - Cavity spot is one of the most common and serious diseases of carrot (Daucus carota L). The disease, caused by different species of Pythium, including P. violae Chesters & Hickman, P. sulcatum Pratt & Mitchell and P. sylvaticum Campbell & Hendrix, leads to frequent high rejection rates during grading worldwide. In the area of the city of Quebec, the disease is caused mainly by P. sulcatum, P. sylvaticum and P. macrosporum Vaartaja & van der Plaats-Niterink. Cavity spot can be controlled with metalaxyl, but reports are emerging that this treatment show little or no efficacy in many regions. This situation reinforces the need for alternative fungicides. The objectives of the present study were: (1) to determine the sensitivity of 14 pathogenic isolates of P. sulcatum, P. sylvaticum and P. macrosporum collected from carrots produced in the area of the city of Quebec to different broad-spectrum and oomycete-specific fungicides (chlorothalonil, etridiazole, fludioxonil, fosetyl-Al, metalaxyl, zoxamide), (2) to evaluate the efficacy of the fungicides in controlling cavity spot, and (3) to evaluate the risk of resistance development of isolates with the best-performing fungicide(s). The determination of EC50 for the fungicides tested showed that most isolates were highly sensitive to both metalaxyl and zoxamide but insensitive to fludioxonil, fosetyl-Al and chlorothalonil. In greenhouse assays, only zoxamide provided significant and consistent disease control as measured by the number of cavity spot lesions caused by P. sulcatum. Investigations into the risk of resistance development to zoxamide showed that, for specific isolates, repeated exposure to the fungicide resulted in a loss of sensitivity. PMID- 15880372 TI - Thyroid cancer and multiple primary tumors in the SEER cancer registries. AB - Thyroid cancer incidence rates have increased steadily in the United States and elsewhere. Radiation exposure at a young age is a strong risk factor, but otherwise the etiology is unclear. To explore etiologic clues, we studied the risk of thyroid cancer after an earlier primary cancer, as well as the risk of developing multiple primaries after an earlier thyroid cancer in the U.S. Surveillance, Epidemiology and End-Results (SEER) cancer registries program (1973 2000). In 2,036,597 patients diagnosed with any invasive cancer who survived for a minimum of 2 months, we observed a 42% increased risk compared to the general population for second thyroid cancer based on 1,366 cases (95% confidence interval (CI) = 35-50%; excess absolute risk (EAR) = 0.38/10,000 person-years (PY)). Elevated risks were observed after most cancer sites studied. The most pronounced excess (observed/expected (O/E) = 2.86) was seen for second thyroid cancers detected in the year after diagnosis of the first cancer. Among 29,456 2 month thyroid cancer survivors, 2,214 second cancers occurred (O/E = 1.11, 95% CI = 1.06-1.15; EAR = 7.64/10,000 PY). Again, the highest risk was seen in the first year (O/E = 1.26). Patients <40 years of age at diagnosis of thyroid cancer had a 39% increased risk of a second cancer, whereas for older patients the risk was elevated 6%. We observed consistently increased risks for cancers of the breast, prostate, and kidney, and a likely radiation treatment-related excess of leukemia. Based on small numbers of cases, cancers of the salivary glands, trachea, scrotum, adrenal glands, and brain and central nervous system (CNS) also occurred in excess. A decreased risk was observed for smoking-related malignancies. Thyroid cancer is associated with primary cancers of many different organs. Although enhanced medical surveillance likely plays a role, 2-way, positive associations between thyroid cancer and cancers of the breast, prostate, kidney, salivary glands, brain and CNS, scrotum, and leukemia suggest etiologic similarities and possible treatment effects. PMID- 15880373 TI - Increased expression of ICAM-3 is associated with radiation resistance in cervical cancer. AB - To search for a marker that predicts the efficacy of radiation therapy in human cervical cancer, gene expression profiles between parental SiHa cervical cancer cells and radiation-resistant SiHa/R cells have been compared by the microarray technique. Microarray and Northern blot analyses demonstrated that the ICAM-3 expression was upregulated in SiHa/R cells. This increased expression of ICAM-3 in SiHa cells enhanced cell survival by about 34.3% after a 2 Gy dosage of radiation. In addition, SiHa/ICAM-3 cells showed a 2.45-fold higher level of FAK phosphorylation than that of the control cells. In tumor specimens, ICAM-3 staining was restricted to tumor stromal endothelial cells and lymphocytes. The overexpression of ICAM-3 was significantly more frequent in radiation-resistant cervical cancer specimens when compared with radiation-sensitive specimens (83.3% vs. 35.3%; p = 0.015). With these observations, we can suggest that an increased expression of ICAM-3 is associated with radiation resistance in cervical cancer cells and the expression of ICAM-3 can be used as a valuable biomarker to predict the radiation resistance in cervical cancer that occurs during radiotherapy. PMID- 15880375 TI - Rainfastness and adsorption of herbicides on hard surfaces. AB - Herbicides are still used to control weeds on hard surfaces, including municipal, private and industrial sites. Used under unfavourable conditions, especially when rain occurs shortly after application, herbicides may run off to surface waters. Such losses of herbicides from hard surfaces are estimated to be much higher than for herbicides used in arable fields. In this study, three kinds of hard surface were evaluated: asphalt, concrete surface and gravel (fine and coarse). Three herbicides were applied: glyphosate, diuron and diflufenican. Adsorption isotherms of diuron and diflufenican to the three surfaces were determined. At different times after treatment with the herbicides, rainfall was simulated by use of a rain-droplet spray nozzle, and the run-off was collected for analysis. After this run-off event, the materials were immersed in water to measure desorption which, together with the compound in the run-off, gave a measure of the dislodgable residues. The apolar herbicides diuron and especially diflufenican adsorbed strongly to asphalt. The polar herbicide glyphosate lost 75% in run-off from asphalt but was adsorbed strongly to soil and concrete pavement. PMID- 15880376 TI - Sildenafil and an early stage of chronic hypoxia-induced pulmonary hypertension in newborn piglets. AB - Devising therapies that might prevent the onset or progression of pulmonary hypertension in newborns has received little attention. Our major objective was to determine whether sildenafil, a selective phosphodiesterase inhibitor, prevents the development of an early stage of chronic hypoxia-induced pulmonary hypertension in newborn pigs. Another objective was to determine whether sildenafil causes pulmonary vasodilation without systemic vasodilation in piglets with chronic pulmonary hypertension. Piglets were raised in room air (control, n = 5) or 10-11% O(2) (hypoxic, n = 17) for 3 days. Some piglets (n = 4) received oral sildenafil, 12 mg/kg/day, throughout exposure to hypoxia. All piglets were anesthetized and catheterized, and pulmonary arterial pressure (Ppa), pulmonary wedge pressure (Pw), aortic pressure (Ao), and cardiac output (CO) were measured. Then for some piglets raised in hypoxia for 3 days, a single oral sildenafil dose (3 mg/kg, n = 6) or placebo (n = 5) was given, and hemodynamic measurements were repeated. For piglets raised in hypoxia for 3 days, mean Ppa and calculated PVR were elevated above respective values in control piglets. Mean Ppa and PVR did not differ between piglets that received sildenafil throughout exposure to hypoxia and those that did not. For piglets with chronic hypoxia-induced pulmonary hypertension that received a single oral dose of sildenafil, mean Ppa and PVR decreased, while mean Pw, CO, mean Ao, and systemic vascular resistance remained the same. All hemodynamic measurements were unchanged after placebo. Oral sildenafil did not influence the early stage of chronic hypoxia-induced pulmonary hypertension in newborn piglets. However, a single oral dose of sildenafil caused pulmonary vasodilation, without systemic vasodilation, in piglets with chronic hypoxia-induced pulmonary hypertension, which may have therapeutic implications. PMID- 15880377 TI - Risk factors of bronchial hyperresponsiveness in children with wheezing associated respiratory infection. AB - The objectives of this study were to identify possible risk factors of bronchial hyperesponsiveness (BHR) in children up to 5 years of age with wheezing associated respiratory infection (WARI), and to study the prevalence of BHR. Children up to 5 years of age with WARI were enrolled in the study. The parents or caregivers of children were asked about their demographic data and clinical histories. Physical examination and clinical score assessment were performed. Pulmonary function tests, i.e., tidal breathing flow volume (TBFV), were performed to measure tidal breathing parameters before and after salbutamol nebulization. If volume at peak tidal expiratory flow/expiratory tidal volume and time to peak expiratory flow/total expiratory time increased > or = 20%, or tidal expiratory flow at 25% of tidal volume/peak tidal expiratory flow increased > or = 20% after nebulization therapy, BHR was diagnosed. The number in the positive BHR group was used to calculate the prevalence of BHR, and clinical features were compared with those of the negative BHR group. Categorical data were analyzed for statistical significance (P < 0.05) by chi-square test or Fisher's exact test, or Student's t-test, as appropriate. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for those with statistical significance. One hundred and six wheezing children underwent pulmonary function tests before and after salbutamol nebulization. With the aforementioned criteria, 41 cases (38.7%) were diagnosed with BHR. History of reactive airway disease, (OR, 6.31; 95% CI, 1.68 25), maternal history of asthma (OR, 3.45; 95% CI, 1.34-9), breastfeeding less than 3 months (OR, 3.18; 95% CI, 1.26-8.12), and passive smoking (OR, 3; 95% CI, 1.15-7.62) were significant risk factors of BHR. The eosinophil count was significantly higher in the BHR (+) group particularly, in children 1-5 years of age (P < or = 0.01). Patchy infiltrates were more commonly found in patients with negative BHR but not statistically significant. In conclusion, a history of reactive airway disease, maternal history, breastfeeding less than 3 months, and passive smoking were significant risk factors for BHR. PMID- 15880378 TI - Enhanced chemotherapy delivery by intraarterial infusion and blood-brain barrier disruption in malignant brain tumors: the Sherbrooke experience. AB - BACKGROUND: The treatment of malignant brain tumors is hampered by the presence of the blood-brain barrier, which limits chemotherapy penetration to the central nervous system (CNS). In recent years, different strategies have been designed to circumvent this physiologic barrier. The osmotic blood-brain barrier disruption (BBBD) procedure is one such strategy, and has been studied extensively in preclinical and clinical studies. The authors detail their experience so far with the procedure in the context of an open Phase II study in the treatment of malignant brain tumors. METHODS: Patients with histologically proven malignant gliomas, primitive neuroectodermal tumors, primary CNS lymphomas, and metastatic disease to the brain were eligible. Patients enrolled were treated every 4 weeks (1 cycle) for < or = 12 cycles. A methotrexate-based regimen was offered to patients with lymphomas, whereas a carboplatin-based regimen was offered to patients with all other histologies. Before intraarterial chemotherapy infusion, patients were submitted to an osmotic BBBD procedure. RESULTS: Seventy-two patients were included in the current report. The overall median survival times (MST) from treatment initiation for glioblastoma multiforme (GBM), anaplastic oligodendrogliomas, primary CNS lymphomas, and metastases were, respectively, 9.1, 13.9, not reached, and 9.9 months, whereas time to disease progression was 4.1, 9.2, 12.3, and 3.3 months. The MST from diagnosis was 32.2 months for GBM. CONCLUSIONS: These encouraging results prompted the authors to further refine their knowledge of the potential contribution of this procedure in the treatment of brain tumors. These authors designed a randomized Phase III study for patients with GBM that is now open. PMID- 15880379 TI - Clinicopathologic and molecular correlations of necrosis in the primary tumor of patients with renal cell carcinoma. AB - BACKGROUND: The presence of histologic necrosis in the primary tumor of patients with renal cell carcinoma (RCC) has been suggested to be an important predictor of survival. The authors investigated the relation of tumor necrosis to other clinicopathologic factors known to be important prognostic indicators for patients with RCC. METHODS: The records of 311 patients undergoing treatment for RCC were evaluated for basic clinicopathologic information including TNM classification, nuclear grade, Eastern Cooperative Oncology Group (ECOG) performance status (PS), disease recurrence, and survival. The presence and extent of histologic necrosis of the primary tumors was recorded and correlated with clinicopathologic factors, carbonic anhydrase IX and Ki-67 expression, disease recurrence, and survival. RESULTS: The presence of necrosis in the primary tumor of patients with RCC compared with patients with RCC without necrosis was associated with higher T classification (P < 0.0001), the presence of lymph node disease (P = 0.009), the presence of metastases (P < 0.0001), higher grade (P < 0.0001), greater mean tumor size (P < 0.0001), an ECOG PS score > or = 1 (P = 0.007), higher University of California-Los Angeles Integrated Staging System (UISS) category (P < 0.0001), and higher Ki-67 expression (P < 0.0001). The extent of necrosis in the primary tumor was associated with the presence of lymph node disease (P = 0.009) and the presence of metastases (P < 0.0001), and correlated with higher T classification (sigma = 0.31, P < 0.0001), poorer ECOG PS (sigma = 0.18, P = 0.002), higher grade (sigma = 0.33, P < 0.0001), greater tumor size (sigma = 0.40, P < 0.0001), higher UISS category (sigma = 0.37, P < 0.0001), and higher Ki-67 staining (sigma = 0.32, P < 0.0001). Patients with the presence of necrosis in the primary tumor demonstrated a lower 5-year disease-specific survival compared with patients without necrosis in the primary tumor (36% vs. 75%; P < 0.0001). Multivariate analysis demonstrated that T classification (P < 0.0001), distant metastases (P < 0.0001), and ECOG PS (P < 0.0001) were independent predictors of DSS, whereas the presence of necrosis was not (P = 0.1100). Substratification into localized and metastatic disease demonstrated that the presence of necrosis was an independent predictor of survival in patients with localized (P = 0.025), but not metastatic (P = 0.44), disease. The extent of necrosis was not an independent predictor of survival (P > 0.05). Patients with the presence of necrosis in the primary tumor had a lower 5 year disease recurrence-free rate compared with patients without the presence of necrosis (62% vs. 92%, P < 0.0001). CONCLUSIONS: The presence of necrosis in the primary tumor was associated with adverse prognostic factors such as high T classification, presence of lymph node disease and metastases, high grade, large tumor size, and poor ECOG PS. The extent of necrosis was found to be associated with the presence of lymph node disease and metastases and correlated with higher T classification, higher grade, greater tumor size, poorer ECOG PS, and higher UISS category. The presence of this histologic variant was an independent predictor of poor survival in patients with localized, but not metastatic, disease. In addition, Ki-67 expression served as a valuable surrogate marker for the presence of histologic tumor necrosis. PMID- 15880380 TI - Frequency of SOX Group B (SOX1, 2, 3) and ZIC2 antibodies in Turkish patients with small cell lung carcinoma and their correlation with clinical parameters. AB - BACKGROUND: Expression of neuroectodermal markers is a key feature of small cell lung carcinoma (SCLC). Although immune responses against a number of these proteins have been associated with paraneoplastic neuronal disease (PND), most patients with SCLC have anti-neuroectodermal antibodies in the absence of PND. Whether these immune responses affect the clinical outcome in SCLC is critical in understanding the potential value of these proteins as cancer vaccine targets as well as in the pathogenesis of PND. METHODS: The authors investigated the frequency of immunoglobulin G autoantibodies against Sry-like high-mobility group box (SOX)1, 2, 3 and Zinc-finger gene of the cerebellum (ZIC)2 proteins in stored serum samples from 90 patients utilizing the lambda-phage plaque assay. Data obtained from patient records were utilized to measure clinical correlates of seroreactivity. RESULTS: Antibodies to SOX1 were present in 28% of patients and another 28% had anti-ZIC2 antibodies, classifying these as some of the most frequent antibody responses observed in SCLC. None had autoimmune paraneoplastic disease. Antibody titers were frequently as high as > or = 1:10(6) and were stable for < or = 6 months after diagnosis. Seroreactivity against either SOX1 or ZIC2 correlated with younger age, lower lactate dehydrogenase levels, and better response to initial therapy. CONCLUSIONS: The frequent and stable presence of SOX Group B and/or ZIC2 antibodies in SCLC, but not in healthy individuals examined, indicates they are serological markers of SCLC. However, the correlation between known clinical parameters of less aggressive disease and seroreactivity suggests that these antibodies are indicators of better prognosis in SCLC and warrants further studies to clarify the nature of the underlying immune responses. PMID- 15880381 TI - A novel method for repetitive peptide synthesis in solution without isolation of intermediates. AB - A novel method was developed for the large-scale manufacture of peptides in solution, called DioRaSSP-Diosynth Rapid Solution Synthesis of Peptides. This method combines the advantages of the homogeneous character of classical solution phase synthesis with the universal character and the amenability to automation inherent to the solid-phase approach. The process consists of repetitive cycles of coupling and deprotection in a permanent organic phase and is further characterized by the fact that intermediates are not isolated. Couplings are mediated by water-soluble carbodiimide. Several types of function may be applied for temporary amino protection depending on the sequence of the actual peptide, including Z, Fmoc, Msc and Nsc. Formate is the preferred hydrogen donor during hydrogenolysis of the Z function, while 1,8-diazabicyclo[5.4.0]undec-7-ene is used to deprotect Fmoc, Msc and Nsc. Morpholine is added during the deprotection of Msc and Nsc to scavenge the arising alkenes. Processes according to this highly efficient synthesis method are easy to scale up and yield products of reproducible high purity, which is guaranteed by a new quenching method for residual activated compounds, applying an anion-forming amine such as a beta alanine ester. This ester should display a lability similar to that of the temporary amino-protecting function, allowing simultaneous deprotection of the growing peptide and the quenched compound. The DioRaSSP approach assures the completely quantitative removal of deprotected quenched compounds before the coupling step of the next cycle of the synthesis by basic aqueous (that is active) extraction, while the growing peptide remains anchored in the organic phase due to the presence of hydrophobic protecting functions. PMID- 15880382 TI - Intrinsic excitability, synaptic potentials, and short-term plasticity in human epileptic neocortex. AB - Although studies of epileptic human hippocampus suggest changes of synaptic and intrinsic excitability, few changes, save the appearance of spontaneous field/synaptic potentials, are known in epileptic neocortical tissue. However, invasive EEG and histological studies suggest that neocortical tissue, even in mesial temporal lobe epilepsy, can play an important role as an irritative zone or extrahippocampal focus. We hypothesized that intrinsic neuronal and synaptic excitability, as well as short-term plasticity, are altered in neocortical areas, particularly with elevated K+ levels as occur during seizures. We analyzed neuronal firing properties, synaptic responses, and paired-pulse plasticity in human neocortical slices from tissue resected during epilepsy surgery, both under normal and under pathological conditions, i.e., after elevating K+ (4/8 mM), with rat neocortical slices as controls. Neuronal firing properties were not different. We did find, however, alterations of synaptic responsiveness in epileptic tissue, i.e., an elevated network excitability with K+ elevations, and reduction of paired-pulse depression. PMID- 15880383 TI - A novel nonsteroidal antifibrotic oligo decoy containing the TGF-beta element found in the COL1A1 gene which regulates murine schistosomiasis liver fibrosis. AB - Schistosomiasis mansoni disseminated worm eggs in mice and humans induce granulomatous inflammations and cumulative fibrosis causing morbidity and possibly mortality. In this study, intrahepatic and I.V. injections of a double stranded oligodeoxynucleotide decoy containing the TGF-beta regulatory element found in the distal promoter of the COL1A1 gene into worm-infected mice suppressed TGF-beta1, COL1A1, tissue inhibitor of metalloproteinase-1, and decreased COL3A1 mRNAs to a lesser extent. Sequence comparisons within the mouse genome found homologous sequences within the COL3A1, TGF-beta1, and TIMP-1 5' flanking regions. Cold competition gel mobility shift assays using these homologous sequences with 5' and 3' flanking regions found in the natural COL1A1 gene showed competition. Competitive gel mobility assays in a separate experiment showed no competition using a 5-base mutated or scrambled sequence. Explanted liver granulomas from saline-injected mice incorporated 10.45 +/- 1.7% (3)H proline into newly synthesized collagen, whereas decoy-treated mice showed no collagen synthesis. Compared with the saline control schistosomiasis mice phosphorothioate double-stranded oligodeoxynucleotide treatment decreased total liver collagen content (i.e. hydroxy-4-proline) by 34%. This novel molecular approach has the potential to be employed as a novel antifibrotic treatment modality. PMID- 15880384 TI - The insulin sensitivity, glucose sensitivity, and acute insulin response to glucose load in adolescent type 2 diabetes in Taiwanese. AB - BACKGROUND: Insulin sensitivity (SI), glucose sensitivity (SG), acute insulin response to glucose load (AIR), and obesity in adolescent type 2 diabetes patients (young diabetes, YDM) in Taiwan were studied. METHODS: Forty patients diagnosed at <22 years of age were enrolled and divided into non-obese (NOYDM, BMI < 27 kg/m(2)) and obese groups (OBYDM BMI > 27 kg/m(2)). Adult-onset type 2 diabetes patients (ADM) >40 years old (n = 41) and nondiabetic young adults (N) (n = 23) served as controls. Fasting plasma lipids, insulin, and glucose were measured. Homeostasis model assessment was calculated to estimate insulin sensitivity and beta-cell function. A frequent-sampled intravenous glucose tolerance test was performed to evaluate SI, SG, and AIR. RESULTS: SI and AIR were significantly lower in YDM and ADM than in N (0.92 +/- 0.13, 0.8 +/- 0.15 and 3.24 +/- 0.47 x 10(-4)/U/mL for SI; 40.3 +/- 20.3, 107.3 +/- 50.2, 1208 +/- 306.3 uU/min for AIR). SG of YDM and ADM were lower compared with N (0.014 +/- 0.00138, 0.0292 +/- 0.0058 vs 0.034 +/- 0.0086 min(-1) respectively). No difference was noted between YDM and ADM. SI and SG were not different in NOYDM and OBYDM. AIR was higher in OBYDM (83.6 +/- 34.3 vs -7.6 +/- 13.66 uU/min). CONCLUSIONS: YDM had defects in SI, SG, and AIR compared to N, which was similar to the pathophysiology of ADM. The results imply that YDM may be either a different subtype of diabetes or the same type of diabetes as ADM, with severe defects associated with earlier age of onset. OBYDM had higher AIR than NOYDM. PMID- 15880385 TI - Design of peptide oxytocin antagonists with strikingly higher affinities and selectivities for the human oxytocin receptor than atosiban. AB - The peptide oxytocin (OT) antagonist atosiban, approved for tocolytic use in Europe (under the tradename Tractocile), represents an important new therapeutic advance for the treatment of premature labor. This paper presents some new peptide OT antagonists which offer promise as superior tocolytics. The solid phase synthesis is reported of four pairs of L and D-2-naphthylalanine (L/D-2Nal) position-2 modified analogs of the following four oxytocin (OT) antagonists: des 9-glycinamide [1-(beta-mercapto-beta,beta-pentamethylene propionic acid), 2-O methyltyrosine, 4-threonine]ornithine-vasotocin (desGly NH(2),d(CH(2))(5)[Tyr(Me)(2),Thr(4)]OVT) (A); the Tyr-NH(2) (9) analog of (A), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Tyr-NH(2) (9)]OVT (B); the Eda(9) analog of (A), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Eda(9)]OVT (C); and the retro COCH(2)Ph(4-0H)(10) modified analog of (C), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Eda(9)<-- COCH(2)Ph(4 0H)(10)]OVT (D). The eight new analogs of A-D are (1) desGly-NH(2),d(CH(2))(5)[D 2Nal(2),Thr(4)]OVT, (2) desGly-NH(2),d(CH(2))(5)[2-Nal(2),Thr(4)]OVT, (3) d(CH(2))(5)[D-2Nal(2),Thr(4),Tyr-NH(2) (9)]OVT, (4) d(CH(2))(5)[2Nal(2),Thr(4),Tyr-NH(2) (9)]OVT, (5) d(CH(2))(5)[D 2Nal(2),Thr(4),Eda(9)]OVT, (6) d(CH(2))(5)[2Nal(2),Thr(4),Eda(9)]OVT, (7) d(CH(2))(5)[D-2Nal(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-0H)(10)]OVT, (8) d(CH(2))(5)[2Nal(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-OH)(10)]OVT. Peptides 1-8 were evaluated for agonistic and antagonistic activities in in vitro and in vivo rat bioassays, in rat OT receptor (rOTR) binding assays and in human OT receptor (hOTR) and human vasopressin (VP) vasopressor (V(1a)) receptor (hV(1a)R) binding assays. Also reported are the hOTR and hV(1a)R affinity data for atosiban and for B. None of the eight peptides exhibit oxytocic or vasopressor agonism. Peptides 1 8 exhibit weak antidiuretic agonism (activities in the range 0.014-0.21 U/mg). Peptides 1-6 exhibit potent in vitro (no Mg(2+)) OT antagonism (anti-OT pA(2) values range from 7.63 to 8.08). Peptides 7 and 8 are weaker OT antagonists. Peptides 1-6 are all OT antagonists in vivo (estimated in vivo anti-OT pA(2) values in the range 6.94-7.23). Peptides 1-8 exhibit vasopressor antagonism, anti V(1a) pA(2) values in the range 5.1-7.65. Peptides 1-8 exhibit high affinities for the rOTR (K(i) values = 0.3-7.8 nM). Peptides 1-4 and B exhibit surprisingly very high affinities for the hOTR; their K(i) values are 0.17, 0.29, 0.07, 0.14 and 0.59 nM, respectively. Peptides 1-4 and B exhibit respectively 449, 263, 1091, 546 and 129 times greater affinity for the hOTR than atosiban (K(i) = 76.4 nM). Peptides 1-4 exhibit high affinities for the hV(1a)R (K(i)s = 1.1 nM, 1.3 nM, 0.19 nM and 0.54 nM, all higher than the hV1(a)R affinities exhibited by atosiban (K(i) = 5.1 nM) and by B (K(i) = 5.26 nM). Because of their strikingly higher affinities for the hOTR than atosiban, peptides 1-4 and B exhibit gains in anti hOT/anti hV(1a) receptor selectivity compared with atosiban of 93, 64, 39, 56 and 127, respectively. These OT antagonists are thus promising candidates for development as potential new tocolytic agents. PMID- 15880386 TI - The stabilizing contribution of thymine in duplexes of (dA)24 with (dU)24, (dT)24, (dU12-dT12), (dU-dT)12, (dU2-dT2)6, or (dU3-dT3)4: nearest neighbor and next-nearest neighbor effects. AB - Ultraviolet melting curves are used to determine the effect of the pyrimidine 5 methyl group on the stability of duplexes of (dA)(24) with (dU)(24), (dT)(24), (dU(12)-dT(12)), (dU-dT)(12), (dU(2)-dT(2))(6), and (dU(3)-dT(3))(4). Substitution of a T for a U results in an increase in stability, which is attributed to an increase in strength of dipole-induced dipole and dispersion (van der Waals) interactions. Significant additional enhancement occurs when two T residues are adjacent. A further increase in the number of adjacent T's has a relatively slight effect on T(m). The sequence effect appears to be largely attributable to an increment in dispersion forces. The CD spectra of the duplexes are all closely similar except in the region between 260 and 290 nm. A band near 272 nm associated with the presence of U in the spectrum of (dA)(24).(dU)(24) decreases in intensity when T's are incorporated in the pyrimidine strand. The band is completely replaced in the spectrum of (dA)(24).(dT)(24) with a new maximum at 282 nm and a minimum at 268 nm, both of lower magnitude. The emergence of the two new bands is correlated with the presence of adjacent T's once more, and only two adjacent T's appear necessary for a major part of the change to occur. The degree of cation release on thermal dissociation of the oligomer dimers ranges from Deltai = 0.14 to 0.16, about the same or slightly less than values reported for polynucleotide duplexes and less than predicted from theoretical calculations. PMID- 15880387 TI - Coping and quality of life among women extensively treated for gynecologic cancer. AB - The current study examined the use of engagement and avoidant coping strategies among advanced-stage gynecologic cancer patients who were facing the long-term stressors of extensive chemotherapy and advanced disease. Patients were compared to a reference group of early-stage patients who had received more limited treatment. Compared to the reference group, extensively-treated women more frequently utilized both engagement and avoidant strategies including active coping, seeking social support, and mental disengagement. Use of engagement coping strategies such as active coping and seeking social support were not significantly related to quality of life or mood. However, avoidant coping strategies, including disengagement and cognitive avoidance, were strongly associated with poorer well-being and more distressed mood. Moreover, relationships between coping and quality of life differed between the extensively treated patients and patients in the limited treatment reference group. The use of avoidance and seeking instrumental support were associated with poorer outcomes among extensively-treated patients but not among the limited treatment group. Results of the current study suggest that coping patterns and outcomes may be more pronounced among cancer patients contending with severe disease and extensive treatment and that avoidant coping strategies may be particularly detrimental with respect to these patients' mood and quality of life. PMID- 15880388 TI - Structural and dynamic properties of cytochrome P450 BM-3 in pure water and in a dimethylsulfoxide/water mixture. AB - Solvent molecules play an important role for the structural and dynamical properties of proteins. A major focus of current protein engineering is the development of enzymes that are catalytically active in the presence of organic solvents. The monooxygenase P450 BM-3 is one of the best-studied enzymes and promising for industrial applications but with limited activity in the presence of organic solvents or cosolvents. To gain insights into the structural and dynamical properties of the heme domain of this enzyme in solution, molecular dynamics simulations in pure water and in a 14% DMSO/water mixture were performed. The results of the simulations show overall similar structural fluctuations in both solvent systems, with no indication of partial or global unfolding. In 14% DMSO, the regions comprising the helices E, F, and the EF loop (implicated in controlling the entry to the active site channel) undergo a large shift. Significant changes were also observed near the active site access channel at the residue R47. During the simulation, no DMSO molecule penetrated the active site. However, a significant accumulation of DMSO molecules close to the substrate-binding site and to the Flavin Mononucleotide (FMN) reductase domain interface was observed. PMID- 15880389 TI - Benefits and costs of methadone treatment: results from a lifetime simulation model. AB - Several studies have examined the benefits and costs of drug treatment; however, they have typically focused on the benefits and costs of a single treatment episode. Although beneficial for certain analyses, the results are limited because they implicitly treat drug abuse as an acute problem that can be treated in one episode. We developed a Monte Carlo simulation model that incorporates the chronic nature of drug abuse. Our model represents the progression of individuals from the general population aged 18-60 with respect to their heroin use, treatment for heroin use, criminal behavior, employment, and health care use. We also present three model scenarios representing an increase in the probability of going to treatment, an increase in the treatment length of stay, and a scenario in which drug treatment is not available to evaluate how changes in treatment parameters affect model results. We find that the benefit-cost ratio of treatment from our lifetime model (37.72) exceeds the benefit-cost ratio from a static model (4.86). The model provides a rich characterization of the dynamics of heroin use and captures the notion of heroin use as a chronic recurring condition. Similar models can be developed for other chronic diseases, such as diabetes, mental illness, or cardiovascular disease. PMID- 15880390 TI - Endoscopic-assisted cranionasal resection of olfactory neuroblastoma. AB - BACKGROUND: Cranionasal resection was first described in 1997 for the surgical resection of olfactory neuroblastoma. The endoscopic transnasal approach is used in cranionasal resection to replace the more invasive craniofacial resection. It has the advantages of avoiding the facial wound and its associated pain, swelling, and scar. The authors have routinely practiced cranionasal resection since 1996 for resection of all anterior skull base tumors in which the resultant skull base bony defect is limited to the nasal and sinus roof. The aim of this study was to review the results of cranionasal resection for olfactory neuroblastoma. METHODS: The results of cranionasal resection for olfactory neuroblastoma in six patients from 1996 to 2003 were reviewed. RESULTS: The Kadesh stages were 3A, 2B, and 1C. None of the patients had postoperative complications. Postoperative radiotherapy was given only to the patient with Kadesh stage C disease. There were no local recurrences. Two patients died of lung metastasis. CONCLUSIONS: Cranionasal resection is a safe and adequate procedure. Postoperative radiotherapy is not necessary after clear resection of Kadesh A and B tumors. PMID- 15880391 TI - Mood stabilization in the treatment of bipolar disorder: focus on quetiapine. AB - The use of at least one mood-stabilizing agent is common clinical practice in the treatment of bipolar disorder, regardless of the treatment setting or disease phase. However, a consensus definition of 'mood stabilizer' remains to be established. A mood stabilizer has been operationally described as an agent that is useful in at least one phase of bipolar disorder while not worsening any other phase of the illness. More stringent definitions have been proposed, and it can be argued that a clinically effective mood stabilizer would have efficacy in a broad range of affective, psychotic, behavioral and cognitive domains in all phases of bipolar disorder and would be well tolerated across a range of doses for sustained periods. Clinically effective mood stabilizers should treat mania and depression, while preventing recurrence and improving quality of life. Effective treatment should not precipitate mania, depression, or rapid cycling, and should minimize the burden of treatment-emergent side effects. Data from clinical studies of quetiapine are reviewed in context with the literature discussing traditional and emerging mood stabilizers. Using a liberal definition, the evidence for quetiapine qualifies it as a bimodal mood stabilizer based on its demonstrated effectiveness in the treatment of bipolar mania and depression. Further data suggest that quetiapine has promise across all phases of bipolar disorder with the potential to meet even the most stringent definitions of a mood stabilizer. PMID- 15880392 TI - Antimicrobial drug use in hospitalised paediatric patients: a cross-national comparison between Germany and Croatia. AB - PURPOSE: To compare the utilisation of systemic antimicrobials at the paediatric units of the university hospitals in Marburg (Germany) and Rijeka (Croatia). METHODS: A prospective, observational analysis of hospital records from 300 incident users of antimicrobials in each study centre that were younger than 19 years. Antimicrobial utilisation was analysed in six gender-specific age groups with respect to drug choice, duration of treatment and hospital stay, indication and route of administration. The extent of antimicrobial drug use was assessed by the number of treatment courses. RESULTS: In each hospital, more than 1/3 of the patients were younger than 1 year. The duration of hospital stay was about two fold longer in Rijeka (18.5 +/- 5.8 days) than in Marburg (8.6 +/- 3.8 days). Pneumonia and other respiratory tract infections were the most common indications in Marburg (38.6%) and Rijeka (58.7%). The cumulative percentage of patients treated with an equal number of different antimicrobials was lower in Rijeka than in Marburg. The most commonly used antimicrobials were ampicillin (40.3%) and cefuroxim (35.9%) in Marburg, but ceftriaxone (43.3%) and cefotaxim (14.0%) in Rijeka. CONCLUSIONS: A shorter treatment duration, less variation in the prescribing pattern and a greater adherence to the use of recommended antimicrobials argue for a more rational antimicrobial drug use in Marburg than in Rijeka. However, a further identification of drug choice determinants is warranted. PMID- 15880393 TI - Plasma cell granuloma of the maxillary sinus: a case report and literature review. AB - BACKGROUND: Plasma cell granulomas are rare and heterogeneous tumor-like lesions of mixed inflammatory cell infiltrates of unknown etiology. Although they have the potential to occur in sites throughout the body, their occurrence in the paranasal sinuses and nasal cavity is uncommon and often associated with unique clinical characteristics and natural history. METHODS: We present a case of an aggressive plasma cell granuloma of the maxillary sinus and a review of the literature (28 cases). RESULTS: The patient was treated with definitive radiotherapy (45 Gy in 25 fractions) and experienced a local recurrence 2 years later. The lesion initially responded to corticosteroids and then progressed. Resection was performed and was followed by another recurrence. The patient was treated with radiosurgery and is disease free 8 years after initial treatment. CONCLUSIONS: Review of the literature indicates that the optimal first line of treatment is high-dose corticosteroids. Surgery is indicated if the lesion fails to respond. Radiotherapy is indicated if complete resection is not feasible. PMID- 15880394 TI - Sclerotherapy to a large cervicofacial vascular malformation: a case report with 24 years' follow-up. AB - BACKGROUND: Large craniofacial venous malformations frequently cause significant cosmetic and functional problems. Treatment of the lesions early in life helps to avoid these problems. We present a case of a large cervicofacial venous malformation. METHODS: The patient was treated with a 5% benzyl alcohol solution of sodium morrhuate. The treatment was begun when the patient was 3 months of age, with a total of 23 injections. RESULTS: The lesion had completely disappeared by age 10. No major complications were observed except a superficial tissue loss on the temporal region. It was healed by conservative treatment. There were no recurrences during the 14-year follow-up. CONCLUSIONS: It is difficult to conclude that this type of treatment is the best choice for patients with venous malformation on the basis of a single case. However, intralesional sclerotherapy should be kept in mind as a saving treatment option in cases of large venous malformations in anatomic regions that present challenges to both the surgeon and the patient. Intralesional sclerotherapy can be performed without serious complications if the sclerosing agent is selected and injected cautiously. PMID- 15880395 TI - Oxaliplatin-induced neurotoxicity and the development of neuropathy. AB - The pathophysiology of oxaliplatin-induced neurotoxicity remains unclear, although in vitro studies suggest involvement of voltage-gated Na+ channels. In the present study, clinical assessment was combined with nerve conduction studies (NCS) and nerve excitability studies in 16 patients after completion of oxaliplatin therapy. Chronic neuropathic symptoms persisted in 50% of patients. NCS confirmed abnormalities in symptomatic patients: sensory potentials were significantly low, whereas motor studies remained essentially normal. At 12-month follow-up of symptomatic patients, positive sensory symptoms improved but NCS abnormalities persisted. Cumulative oxaliplatin dose was a predictor of neuropathy, and long-term effects appeared to be minimized by low single-infusion dosages. Nerve excitability measures in symptomatic patients established that axons were of high threshold. Refractoriness was significantly greater in patients (symptomatic group, 56.3 +/- 24.9%; entire patient group, 46.3 +/- 12.5%; controls, 27.1 +/- 1.9%; P < 0.05). Thus, although positive sensory symptoms of oxaliplatin-induced neuropathy improved, negative sensory symptoms and abnormalities of sensory nerve conduction persisted. Differences in nerve excitability measures, particularly refractoriness, support in vitro studies indicating involvement of voltage-gated transient Na+-channel dysfunction in the development of oxaliplatin-induced neurotoxicity. PMID- 15880396 TI - The impact of influent nutrient ratios and biochemical reactions on oxygen transfer in an EBPR process--a theoretical explanation. AB - In this investigation, a laboratory-scale enhanced biological phosphorus removal (EBPR) process was operated under controlled conditions to study the impact of varying the influent ratio of chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN) and total phosphorus (TP), and the consequential biochemical reactions on oxygen transfer parameters. The data showed that the experiment with high influent phosphorus relative to nitrogen (COD/TP = 51 and TKN/TP = 3.1) achieved higher alpha and oxygen transfer efficiency (OTE(f)). On the other hand, the experiment with high influent nitrogen relative to phosphorus (TKN/TP = 14.7 and COD/TP = 129) resulted in approximately 50% reduction in alpha and OTE(f) under similar organic loading. This suggested that the intracellular carbon storage and the enhanced biological P removal phenomenon associated with the phosphorus-accumulating organisms (PAOs) had a positive influence on OTE(f) in the high phosphorus experiment compared to an active population of nitrifying and denitrifying organisms in the high nitrogen experiment. The intracellular carbon storage by the glycogen-accumulating organisms also appeared to have had a positive effect on oxygen transfer efficiency, although to a lesser extent in comparison to the PAOs. It was also found that oxygen uptake rate (OUR) was not a good indicator of the measured alpha and OTE(f), because it was a combined effect of several biochemical reactions, each having a varying degree of influence. It is difficult to underestimate the crucial role of flocs in mass transfer of oxygen, because microorganisms associated with flocs carry out the biochemical reactions. It seems that the combination of influent characteristics and biochemical reactions in each experiment produced a unique biomass quality (determined by the biomass N to P ratio), ultimately affecting the mass transfer of oxygen. A theoretical explanation for the observed oxygen transfer efficiency under the process conditions is also proposed in this article. PMID- 15880397 TI - A review of the possible relevance of inositol and the phosphatidylinositol second messenger system (PI-cycle) to psychiatric disorders--focus on magnetic resonance spectroscopy (MRS) studies. AB - Myo-inositol is an important part of the phosphatidylinositol second messenger system (PI-cycle). Abnormalities in nerve cell myo-inositol levels and/or PI cycle regulation has been suggested as being involved in the pathophysiology and/or treatment of many psychiatric disorders including bipolar disorder, major depressive disorder, panic disorder, obsessive-compulsive disorder, eating disorders and schizophrenia. This review examines the metabolism and biochemical importance of myo-inositol and the PI-cycle. It relates this to the current in vivo evidence for myo-inositol and PI-cycle involvement in these psychiatric disorders, particularly focusing upon the magnetic resonance spectroscopy (MRS) findings in patient studies to date. From this review it is concluded that while the evidence suggests probable relevance to the pathophysiology and/or treatment of bipolar disorder, there is much less support for a significant role for the PI cycle or myo-inositol in any other psychiatric disorder. More definitive investigation is required before PI-cycle dysfunction can be considered specific to bipolar disorder. PMID- 15880398 TI - Potential anti-cancer effects of virgin olive oil phenols on colorectal carcinogenesis models in vitro. AB - The traditional Mediterranean diet is thought to represent a healthy lifestyle; especially given the incidence of several cancers including colorectal cancer is lower in Mediterranean countries compared to Northern Europe. Olive oil, a central component of the Mediterranean diet, is believed to beneficially affect numerous biological processes. We used phenols extracted from virgin olive oil on a series of in vitro systems that model important stages of colon carcinogenesis. The effect the extract on DNA damage induced by hydrogen peroxide was measured in HT29 cells using single cell microgel-electrophoresis. A significant anti genotoxic linear trend (p=0.011) was observed when HT29 cells were pre-incubated with olive oil phenols (0, 5, 10, 25, 50, 75, 100 microg/ml) for 24 hr, then challenged with hydrogen peroxide. The olive oil phenols (50, 100 microg/ml) significantly (p=0.004, p=0.002) improved barrier function of CACO2 cells after 48 hr as measured by trans-epithelial resistance. Significant inhibition of HT115 invasion (p<0.01) was observed at olive oil phenols concentrations of 25, 50, 75, 100 microg/ml using the matrigel invasion assay. No effect was observed on HT115 viability over the concentration range 0, 25, 50 75, 100 microg/ml after 24 hr, although 75 and 100 microg/ml olive oil phenols significantly inhibited HT115 cell attachment (p=0.011, p=0.006). Olive oil phenols had no significant effect on metastasis-related gene expression in HT115 cells. We have demonstrated that phenols extracted from virgin olive oil are capable of inhibiting several stages in colon carcinogenesis in vitro. PMID- 15880399 TI - A weighted cohort approach for analysing factors modifying disease risks in carriers of high-risk susceptibility genes. AB - The authors propose a novel approach to evaluate the effects of risk factors on disease risks in carriers of high-penetrance alleles in disease susceptibility genes. Most studies to date have utilised data collected on carriers identified through ongoing genetic testing programs. The advantage of this approach is that it allows relatively large numbers of affected and unaffected carriers to be identified rapidly. However, genetic testing is targeted at individuals with a strong family history of disease, so that the selection of carriers is not random with respect to disease status. Risk factors are often analysed by standard cohort analysis methods, but these can be biased in retrospective studies if subjects are selected on the basis of phenotype. To overcome this problem, a weighted cohort approach is proposed, under which individuals are weighted according to certain sampling probabilities in order to mimic a true cohort. The method is illustrated by analyses of data from the International BRCA1/2 Carrier Cohort Study (IBCCS). Simulations demonstrate that the method gives rate ratio estimates that are close to unbiased provided that the absolute disease risks are well estimated. The power to detect associations is, however, reduced compared with an unweighted approach. PMID- 15880400 TI - The HIV-Tat protein induces chromosome number aberrations by affecting mitosis. AB - To analyze the effects of the HIV-Tat-tubulin interaction, we microinjected HIV Tat purified protein into Drosophila syncytial embryos. Following the Tat injection, altered timing of the cortical nuclear cycles was observed; specifically, the period between the nuclear envelope breakdown and anaphase initiation was lengthened as was the period between anaphase initiation and the formation of the next nuclear envelope. These two periods correspond to kinetochore alignment at metaphase and to mitosis exit, respectively. We also demonstrated that these two delays are the consequence of damage specifically induced by Tat on kinetochore alignment and on the timing of sister chromatid segregation at anaphase. Furthermore, we show that the expression of Tat in Drosophila larvae brain cells produces a significant percentage of polyploid and aneuploid cells. The results reported here indicate that Tat impairs the mitotic process and that Tat-tubulin interaction appears to be responsible for the observed defects. The presence of polyploid and aneuploid cells is consistent with a delay or arrest in the M phase of a substantial fraction of the cells expressing Tat, suggesting that mitotic spindle checkpoints are overridden following Tat expression. PMID- 15880401 TI - Disordered respiratory control in children with partial cerebellar resections. AB - While the cerebellum is not traditionally thought of as having an important role in respiratory control, breathing involves cyclic motor acts that require cerebellar coordination. We postulate that children with partial cerebellar resections have disordered respiratory control due to altered synchronization of ventilatory muscles. We reviewed the records of 36 children following partial cerebellar resections due to neoplasms confined to the cerebellum. P aCO2 values were elevated in 19% of patients. Six patients had apneic or bradypneic events documented within the first month after resection. Two patients required intubation with assisted ventilation, and one needed assisted ventilation for 7.3 weeks. Those with apnea had lower oxygen saturations, and a longer need for supplemental oxygen. Patients with apnea were older than those without apnea. Swallowing, which uses many of the same muscles as those needed to maintain upper airway patency, was dysfunctional in 50% of those with apneas. We conclude that children with cerebellar resections have an increased incidence of apnea, hypoventilation, and hypoxemia not otherwise explained by pulmonary disease, and some require prolonged assisted ventilation. We speculate that these abnormalities are manifestations of altered respiratory control caused by dysfunctional cerebellar coordination of ventilatory muscles. PMID- 15880402 TI - Bronchiolitis obliterans organizing pneumonia as an initial manifestation in systemic lupus erythematosus. AB - Bronchiolitis obliterans organizing pneumonia (BOOP) is a rare complication of adult systemic lupus erythematosus (SLE). This is the first report of a pediatric patient with BOOP as an initial presentation of SLE. She had dyspnea, cough, arthralgia, and erythema on her face. Laboratory examinations revealed pancytopenia, low serum levels of complements, and positivity for anti-nuclear antibody, anti-double stranded DNA antibody, and anti-SM antibody. Her respiratory symptoms, pulmonary function tests, and radiologic findings showed significant improvement after treatment with oral prednisolone. Although it is a rare complication among the pleuro-pulmonary manifestations in SLE, BOOP can be the first presentation, even in pediatric patients. PMID- 15880403 TI - Acoustic arousal responses in children with obstructive sleep apnea. AB - Our objectives were to study the arousal responses to nonrespiratory (acoustic) stimuli in children with obstructive sleep apnea syndrome (OSAS). The acoustic arousal response was studied in children with OSAS due to adenotonsillar hypertrophy compared to normal, age-matched children. Acoustic stimuli were delivered incrementally from 30-100 dB during stage 2, slow wave sleep, and rapid eye movement (REM) sleep. The percentage of children who aroused in response to acoustic stimuli, and the arousal threshold (i.e., sound level at which arousal occurred), were compared between groups and sleep stages. The percentage of children who aroused was similar between children with OSAS and controls. The percentage of children who aroused was lower during slow wave sleep than REM sleep and stage 2 in both OSAS and controls. There were no statistically significant differences in acoustic arousal threshold between OSAS and control children. There was no difference in arousal response to moderate acoustic stimulation between children with OSAS and controls. These results contrast with previous data showing blunted arousal responses to hypercapnia and upper airway loading during sleep in children with OSAS, suggesting that children with OSAS have an arousal deficit specific to respiratory stimuli. However, further studies evaluating arousal to both respiratory and nonrespiratory stimuli in the same subjects are needed. PMID- 15880404 TI - Expression of all Wnt genes and their secreted antagonists during mouse blastocyst and postimplantation development. AB - In this extensive study, real-time reverse transcriptase-polymerase chain reaction was used to analyze the expression levels of all 19 Wnt genes and their 11 potential antagonists in mouse blastocysts, pregastrula, gastrula, and neurula stages. By complementing these results with in situ hybridization, we revealed new expression domains for Wnt2b and Sfrp1, respectively, in the future primitive streak at the posterior side and in the anterior visceral endoderm before the initiation of gastrulation. Moreover, the anterior visceral endoderm expresses three secreted Wnt antagonists (Sfrp1, Sfrp5, and Dkk1) in partially overlapping domains. We also identified expression patterns for the Wnt1, Wnt3a, Wnt6, Wnt7b, Wnt9a, Wnt10b, and Sfrp1 genes at the blastocyst stage. In particular, the expression of Wnt1 and Sfrp1 predominantly in the inner cell mass and of Wnt9a in the mural trophoblast and inner cell mass cells surrounding the blastocoele suggests new roles for the Wnt pathway in preimplantation development. This article is the first report on the regional expression of Wnt genes in the mouse blastocyst. PMID- 15880405 TI - LDL-apheresis improves peripheral arterial occlusive disease with an implication for anti-inflammatory effects. AB - Although it is known that LDL-apheresis improves ischemic limb seen in patients with peripheral arterial occlusive disease (PAOD), anti-inflammatory effects are not well known. We studied whether or not serum or plasma levels of high sensitivity C-reactive protein (hsCRP), monocyte chemoatractant protein-1 (MCP 1), or fibrinogen could contribute to favorable effects for ischemic limbs after LDL-apheresis. Twenty-eight patients with PAOD (24 men, 4 women) were enrolled in our study. LDL-apheresis was performed 10 times (treated plasma of 3,000 ml) for 5 weeks. Serum levels of logarithmically transformed values of hsCRP significantly decreased from 3.666 +/- 0.126 to 3.482 +/- 0.139 ng/ml before and after a single session of LDL-apheresis (P < 0.001). Serum levels of MCP-1 decreased from 233 +/- 17.5 to 187 +/- 13.5 pg/ml before and after LDL-apheresis (P < 0.05). Likewise, plasma fibrinogen levels statistically decreased from 196 +/- 9.82 to 159 +/- 9.60 mg/dl (P < 0.001). Overall rates of improvement including foot chillness or numbness, and double folds increase in walking distance were 82.1% 3 months after a completion of LDL-apheresis, while gangrene was only improved 14.3%. Intermittent claudication improved in 53.6%. The favorable actions of LDL-apheresis might include anti-inflammatory effects. To avoid amputation, LDL-apheresis should be applied for patients with PAOD at an early stage of the disease process and may be applicable for patients with atherosclerotic cardiovascular disorders. PMID- 15880406 TI - Apheresis for MPO-ANCA-associated RPGN-indications and efficacy: lessons learned from Japan nationwide survey of RPGN. AB - A national survey concerning rapidly progressive glomerulonephritis (RPGN) was conducted in Japan between 1989 and 2000 and resulted in the registration of 715 patients with RPGN. Among the documented patients, the most frequent primary disease was primary pauci-immune crescentic glomerulonephritis (n = 283), and the second most frequent was microscopic polyangitis (n = 127). Overall, 370 patients had MPO-ANCA, and 23 patients had PR3-ANCA. We found that both renal and patient survivals were significantly worse in patients with MPO-ANCA-associated RPGN than patients with PR3-ANCA. Fifty-three patients received apheresis therapy with various combinations of immunosuppressive regimens. They had higher serum creatinine, higher CRP, and a higher frequency of complicated pulmonary involvements as compared to the controls without apheresis therapy. In dialysis dependent patients, no additional benefit from apheresis therapy was observed. Only pulmonary renal syndrome patients with CRP > 6 mg/dl at presentation showed a slightly better prognosis (patient survival with apheresis; 66.7%, without apheresis; 56.7%). Furthermore, a rapid MPO-ANCA titer reduction was observed in patients treated with apheresis. Patients with MPO-ANCA-associated RPGN were older, and had more chronic and sclerotic lesions than patients with PR3-ANCA associated RPGN. Based on these findings, we suggest that a lower dose of immunosuppressant should be considered in order to avoid opportunistic infection. In this situation, cytapheresis is the treatment of choice. Nevertheless, in patients with an aggressive form of RPGN with rapid deterioration of renal function like the PR3-ANCA-associated RPGN, or pulmonary renal syndrome complicated severe inflammation, or relapses with high MPO-ANCA titer, we conclude that apheresis therapy should be considered. PMID- 15880407 TI - Surveyor Nuclease: a new strategy for a rapid identification of heteroplasmic mitochondrial DNA mutations in patients with respiratory chain defects. AB - Molecular analysis of mitochondrial DNA (mtDNA) is a critical step in diagnosis and genetic counseling of respiratory chain defects. No fast method is currently available for the identification of unknown mtDNA point mutations. We have developed a new strategy based on complete mtDNA PCR amplification followed by digestion with a mismatch-specific DNA endonuclease, Surveyor Nuclease. This enzyme, a member of the CEL nuclease family of plant DNA endonucleases, cleaves double-strand DNA at any mismatch site including base substitutions and small insertions/deletions. After digestion, cleavage products are separated and analyzed by agarose gel electrophoresis. The size of the digestion products indicates the location of the mutation, which is then confirmed and characterized by sequencing. Although this method allows the analysis of 2 kb mtDNA amplicons and the detection of multiple mutations within the same fragment, it does not lead to the identification of homoplasmic base substitutions. Homoplasmic pathogenic mutations have been described. Nevertheless, most homoplasmic base substitutions are neutral polymorphisms while deleterious mutations are typically heteroplasmic. Here, we report that this method can be used to detect mtDNA mutations such as m.3243A>G tRNA(Leu) and m.14709T>C tRNA(Glu) even when they are present at levels as low as 3% in DNA samples derived from patients with respiratory chain defects. Then, we tested five patients suffering from a mitochondrial respiratory chain defect and we identified a variant (m.16189T>C) in two of them, which was previously associated with susceptibility to diabetes and cardiomyopathy. In conclusion, this method can be effectively used to rapidly and completely screen the entire human mitochondrial genome for heteroplasmic mutations and in this context represents an important advance for the diagnosis of mitochondrial diseases. PMID- 15880408 TI - Osteoblastoma of the mandible: clinicopathologic study of four cases and literature review. AB - BACKGROUND: Osteoblastoma is a benign bone tumor accounting for 1% of all bone tumors; it commonly involves the spine and the sacrum of young individuals, with less than 5% being localized to the posterior mandible. In view of its rarity in the maxilla and mandible, osteoblastoma is rarely diagnosed as such in the absence of interdisciplinary cooperation. METHODS: A retrospective study of four benign osteoblastomas was performed based on a review of the clinical, radiographic, and histopathologic features of all cases. RESULTS: The tumors involved the posterior mandible of young patients (age range, 10-21 years; two male and two female patients) and appeared as painful bone expansions. Radiologically, they were poorly defined, radiolucent/radiopaque lesions containing calcifications and not showing sclerotic borders or periosteal reactions. Histologically, they were composed of osteoid and woven bone surrounded by plump osteoblast-like cells with interposed fibroblasts, inflammatory cells, and red blood cells. All patients were disease free after prolonged follow-up. CONCLUSIONS: Osteoblastomas may be distinguished from other bone tumors, fibro-osseous lesions, and odontogenic neoplasms on the basis of integrated clinical, radiologic, and histologic features and usually manifest an indolent clinical course. PMID- 15880409 TI - Proliferation in the epidermis of chelonians and growth of the horny scutes. AB - The proliferation of the epidermis in soft skin, claws, and scutes of the carapace and plastron in the tortoise (Testudo hermanni) and the turtle (Chrysemys picta) were studied using autoradiographic and immunocytochemical methods. During the growing season, a basal keratinocyte in the epidermis of soft skin and claws takes 5-9 days to migrate into the corneous layer. In the tortoise, during fall/winter (resting season) a few alpha-keratin cells are produced in soft epidermis and hinge regions among scutes and occasional beta keratin cells in the outer scute surface. When growth is resumed in spring (growing season), cell proliferation is intense, mainly around hinge regions and tips of marginal scutes. No scute shedding occurs and numerous beta-keratin cells are produced around the hinge regions, while alpha-keratin cells disappear. Beta cells form a new thick corneous layer around the hinge regions, which constitute the growing rings of scutes. Beta-keratin cells produced in more central parts of scutes maintain a homogeneous thickness of the corneous layer along the whole scute surface. In the turtle, a more complicated process of scute growth occurs than in the tortoise. At the end of the growing season (late fall) the last keratinocytes formed beneath the old stratum corneum of the outer scale surface and hinge regions produce more alpha- than beta-keratin. These thin alpha-keratin cells form a scission layer below the old stratum corneum, which extends from the hinge regions toward the center of scutes and the tip of marginal scutes. In the resting season (fall/winter) most cells remain within the germinative layer of the carapace and plastron and a few alpha-cells move in 7-9 days into the corneous layer above hinge regions. In the following spring/summer (growing season) a new generation of beta-keratin cells is produced beneath the scission layer from the hinge region and more central part of the scutes. The epidermis of the inner surface of scutes and hinge regions contains most of the cells incorporating thymidine and histidine, while the remaining outer scute surface is less active. It takes 5-9 days for a newly produced beta-cell to migrate into the corneous layer. These cells form a new corneous layer that extends the whole scute surface underneath the maturing scission layer. The latter contains lipids and eventually flakes off, determining shedding of the above outer corneous layer in late spring or summer. PMID- 15880410 TI - Treatment results of 1070 patients with nasopharyngeal carcinoma: an analysis of survival and failure patterns. AB - BACKGROUND: The aim of this analysis was to evaluate the outcomes of patients with nasopharyngeal carcinoma (NPC) treated primarily by external beam irradiation (ERT) and to explore for possible ways to improve the treatment results. METHODS: One thousand seventy patients with nonmetastatic NPC treated from 1990 to 1998 were retrospectively analyzed. The distribution according to the Union Internationale Contre le Cancer (UICC) (1997 edition) staging system at initial diagnosis was as follows: stage I, n = 113; stage IIA, n = 38; stage IIB, n = 360; stage III, n = 306; stage IVA, n = 136; stage IVB, n = 117; T1, n = 284; T2a, n = 88; T2b, n = 398; T3, n = 149; T4, n = 151; N0, n = 321; N1, n = 393; N2, n = 238; N3a, n = 29; N3b, n = 89. Two hundred eight patients were given neoadjuvant chemotherapy. Ninety-seven patients were diagnosed with locally persistent disease and were salvaged with high dose rate intracavitary brachytherapy. Multivariate analysis was performed with the Cox regression proportional hazards model. RESULTS: The 5-year actuarial local failure-free survival, regional failure-free survival, distant metastasis-free survival, progression-free survival, cancer-specific survival, and overall survival rates were 80.9%, 93.3%, 77.2%, 62.7%, 71.4%, and 66.5%, respectively. Isolated distant metastasis occurred in 191 patients (18%). The distributions were as follow: stage I, 2.1% (two of 95); stage IIA, 5.7% (two of 35); stage IIB, 14.9% (45 of 302); stage III, 26.4% (62 of 235); stage IVA, 40% (40 of 100); stage IVB, 47.1% (40 of 85). Results of the multivariate analysis of various clinical endpoints were discussed. By studying these failure patterns, it is hoped that we could refine future treatments according to the failure patterns of patients with different risks of locoregional and distant failure. CONCLUSIONS: The 18% incidence of isolated distant metastasis is too high to be ignored. Maximizing the local control and minimizing the risk of distant metastasis and late complications should be the key objectives in designing future clinical trials. PMID- 15880411 TI - Long-term survival in locally advanced oral cavity cancer: an analysis of patients treated with neoadjuvant cisplatin-based chemotherapy followed by surgery. AB - BACKGROUND: Neoadjuvant chemotherapy has been reported to be extremely active in head and neck cancer but has failed to give a statistically significant improvement in survival. METHODS: From 1981 to 1994, 33 operable patients with locally advanced oral cavity cancer received cisplatin-based chemotherapy before surgery. Postoperative radiotherapy was performed in high-risk patients. RESULTS: The overall clinical and pathologic complete response rates to neoadjuvant chemotherapy were 48% and 30%, respectively. At a median follow-up of 7.0 years (range, 0.3-15.3+ years), the 5-year and 10-year overall survival rates were 54.5% and 39.5%, and the disease-specific median survival was 6.6 years for all patients (8.3 and 2.3 years for stages III and IV, respectively). The univariate analysis showed a positive relationship between survival and male sex (p = .05), pathologic (p = .02), and clinical (p = .03) complete response. The Cox proportional hazard regression model confirmed the independent prognostic value of the clinical response with a 4.67 (95% CI, 1.70-12.86) hazard ratio. A second primary tumor occurred in six patients (18%), with a median of occurrence of 9 years (range, 7-11 years). CONCLUSIONS: This study confirms the prolonged survival expectancy largely exceeding 5 years for selected patients with stage IV and for most with stage III locally advanced oral cavity cancer achieving a clinical and/or pathologic complete response to chemotherapy. PMID- 15880412 TI - Protein abnormality in denervated skeletal muscles from patients with brachial injury. AB - A proteomic analysis was performed to compare protein expression between normal sternocleidomastoid muscle and denervated muscle. Two-dimensional electrophoresis (2-DE) of muscle proteins showed that 26 proteins among about 800 spots in 2-DE gel displayed a decrease and 6 proteins an increase in expression in muscles with denervation atrophy compared to normal controls; the identified proteins that were abnormally expressed could be generally grouped together as metabolic proteins, chaperone proteins, and contractile-apparatus proteins. The significance of these altered proteins is discussed. In particular, the decrease in hD54 may reduce the activity of transmembrane signaling in atrophied muscle, while the disregulation of DnaJC 1 showed a possible role of molecular chaperones in the functional recovery of atrophied muscles. PMID- 15880413 TI - Minimal access therapy in pediatric pulmonary hydatid cysts. AB - Hydatid cysts occur commonly in lungs in the pediatric age group. Though medical management has been tried, definitive management of the condition is essentially surgical. Use of video-assisted thoracic surgery (VATS) has been rarely employed for the management of pulmonary hydatid cysts. We present a 10-year-old boy with pulmonary hydatid cysts treated with VATS-assisted minithoracotomy with hydatid cyst enucleation and capitonnage. The patient developed a lung abscess postoperatively and had to undergo lobectomy. The surgical management of pulmonary hydatid cysts is discussed here. The role of minimal access therapy for pulmonary hydatid cysts is highlighted. PMID- 15880414 TI - Role of smoking in global and regional cancer epidemiology: current patterns and data needs. AB - Although smoking is widely recognized as a major cause of cancer, there is little information on how it contributes to the global and regional burden of cancers in combination with other risk factors that affect background cancer mortality patterns. We used data from the American Cancer Society's Cancer Prevention Study II (CPS-II) and the WHO and IARC cancer mortality databases to estimate deaths from 8 clusters of site-specific cancers caused by smoking, for 14 epidemiologic subregions of the world, by age and sex. We used lung cancer mortality as an indirect marker for accumulated smoking hazard. CPS-II hazards were adjusted for important covariates. In the year 2000, an estimated 1.42 (95% CI 1.27-1.57) million cancer deaths in the world, 21% of total global cancer deaths, were caused by smoking. Of these, 1.18 million deaths were among men and 0.24 million among women; 625,000 (95% CI 485,000-749,000) smoking-caused cancer deaths occurred in the developing world and 794,000 (95% CI 749,000-840,000) in industrialized regions. Lung cancer accounted for 60% of smoking-attributable cancer mortality, followed by cancers of the upper aerodigestive tract (20%). Based on available data, more than one in every 5 cancer deaths in the world in the year 2000 were caused by smoking, making it possibly the single largest preventable cause of cancer mortality. There was significant variability across regions in the role of smoking as a cause of the different site-specific cancers. This variability illustrates the importance of coupling research and surveillance of smoking with that for other risk factors for more effective cancer prevention. PMID- 15880415 TI - Switch of HLA-G alternative splicing in a melanoma cell line causes loss of HLA G1 expression and sensitivity to NK lysis. AB - Considerable information has been accumulated on HLA-G expression in tumor lesions in which HLA-G is viewed as a way to turn off anti-tumoral immunity. Nevertheless, there is little data concerning the mechanisms by which expression and function of HLA-G are regulated in malignant cells. Here, we have addressed these points by studying a melanoma cell line derived from a surgically-removed HLA-G-positive melanoma lesion. We show that HLA-G expression in melanoma cells can be regulated at the mRNA splicing level. Indeed, melanoma cells rapidly switched from cell-surface HLA-G1 to intra-cellular HLA-G2 expression. This mechanism restored tumor sensitivity to NK lysis. Moreover, switch from HLA-G1 to HLA-G2 was strong enough to prevent re-expression of immunoprotective HLA-G1 even following treatments with cytokines and DNA demethylating agent. Modulating HLA-G at the mRNA splicing level would be an efficient way of lifting in vivo HLA-G mediated tumor immune escape. PMID- 15880416 TI - Micronucleus test in fish cells: a bioassay for in situ monitoring of genotoxic pollution in the marine environment. AB - To evaluate the use of native fish species for assessing genotoxic pollution in the marine environment, micronucleus (MN) analysis was performed in peripheral blood erythrocytes and gill cells of the grey mullet (Mugil cephalus) from three sampling stations off the southeastern Mediterranean coast of Turkey. The frequencies of blebbed, notched, and lobed nuclei and binucleated cells also were evaluated in peripheral erythrocytes. The sampling sites were chosen on the basis of pollution levels; Karaduvar harbor, contaminated by different types of industrial effluents, and Mersin harbor, mainly contaminated by aromatic hydrocarbons, were selected as polluted areas. Erdemli harbor, a relatively unpolluted site, was used as the control area. Sampling was carried out at four different seasons. The frequencies of both micronuclei and other nuclear abnormalities (NAs) in mullets captured from polluted areas were significantly higher than those in mullets from the reference area. In general, gill cells had considerably higher MN frequencies than did erythrocytes, and genotoxic responses were higher in summer than in winter. The results of this study indicate that the MN test in fish is a suitable biomarker for in situ monitoring of genotoxic pollution in the marine environment. As demonstrated in this study, NAs other than micronuclei are also useful indices of chemical exposure and toxic responses. Therefore, measuring both micronuclei and NAs may increase the sensitivity of the test system. PMID- 15880417 TI - Oral rehabilitation after treatment for head and neck malignancy. AB - BACKGROUND: Advances in the management of oral malignancy have resulted in significant improvements in survival and functional outcome. Ablation of oral tissues and radiotherapy render many patients unable to wear conventional prostheses, and these patients are, thus, candidates for oral rehabilitation with osseointegrated implants. We aim to present outcomes and complications of such treatment over a 14-year period in a single unit. METHODS: Data were collected for 81 consecutive patients, most of whom had received microvascular free flap reconstruction after surgical ablation of oral squamous cell carcinoma. Three hundred eighty-six implants were placed after a delay of 12 months after surgery. Sixty-five percent of implants were placed in the anterior mandible. Radiotherapy was used in 47% of the patients, and hyperbaric oxygen treatment was routinely used in irradiated subjects during the latter half of the series. Retrospective analysis of implants and prostheses was made by use of case notes, radiographs, and a computerized database. RESULTS: Data are presented for 364 of the 386 implants in 77 of the 81 patients after a median follow-up of 4 years. Two hundred sixty-five (73%) of the implants were in function supporting prostheses, 56 (15%) had been lost, and 43 (12%) were present but not loaded (ie, "sleepers"). Implant loss seemed patient specific and was also correlated with host bone type. Thirteen percent of patients in whom implants were placed in the mandible lost at least one implant, and the equivalent values for the maxilla was 40%. Thirty-six percent of patients in whom implants were placed in bone graft or flap lost at least one implant. The effects of implant manufacture, dimensions, radiotherapy, and hyperbaric oxygen did not reach statistical significance in this series. Cases of a second primary malignancy were noteworthy; however, the impact of recurrence was minimized by the delay between resection and rehabilitation. Of the 42 fixed and 29 removable prostheses fitted, 12 (17%) failed. CONCLUSIONS: Implants placed in mandible were reliable, but failure rates in vascularized bone graft and maxilla were higher. Radiotherapy did not seem to prejudice implant survival, and hyperbaric oxygen had no demonstrable benefit in this series. Despite some persistent soft tissue problems and implant loss, most patients reached a successful prosthetic and functional outcome. PMID- 15880418 TI - Salvage of free second toe with venous thrombosis by shunting retrograde arterial flow of second dorsal metatarsal artery. AB - Although there are case reports about salvaging amputated digits without venous return by shunting retrograde blood flow from a distal digital artery to a proximal vein (Smith et al., Plast Reconstr Surg 71:52, 1983), successful salvage of a congested free toe with efferent arteriovenous (AV) shunting retrograde arterial flow from the second dorsal metatarsal artery (SDMA) has never been reported. In this paper, we describe the possible factors for venous thrombosis, the errors we committed in the operation, and how the free toe can survive without venous drainage. PMID- 15880419 TI - Role of mitogen-activated protein kinases in phenethyl isothiocyanate-induced apoptosis in human prostate cancer cells. AB - The present study was undertaken to examine the role of mitogen-activated protein kinases (MAPKs) in apoptosis induction by phenethyl isothiocyanate (PEITC), a cruciferous vegetable-derived cancer chemopreventive agent, with DU145 and LNCaP human prostate cancer cells as a model. The MAPK family of serine/threonine kinases, including extracellular signal-regulated kinase1/2 (ERK1/2), c-jun N terminal kinase1/2/3 (JNK1/2/3), and p38 MAPK play an important role in cell proliferation and apoptosis in response to different stimuli. Exposure of DU145 and LNCaP cells to growth suppressive concentrations of PEITC resulted in activation of ERK1/2 and JNKs, but not p38 MAPK, in both cell lines. In DU145 cells, the apoptosis induction by PEITC was statistically significantly attenuated by pharmacological inhibition of JNKs with SP600125. Adenovirus mediated overexpression of Flag-tagged JNK binding domain (JBD) of JNK interacting protein-1 (JIP-1), an inhibitor of JNK, also inhibited PEITC-induced apoptosis in DU145 cells. On the other hand, inhibition of ERK1/2 activation with MEK1 inhibitor PD98059 failed to offer protection against PEITC-induced apoptosis in DU145 cells. In LNCaP cells, the PEITC-induced cell death was not affected by either pretreatment with PD98059 or SP600125 or overexpression of JBD of JIP-1. These results indicate that involvement of MAPKs in apoptosis induction by PEITC in human prostate cancer cells is cell line-specific. PMID- 15880420 TI - Sweat-testing in preterm and full-term infants less than 6 weeks of age. AB - Our objective was to examine the characteristics of preterm and full-term infants < or = 6 weeks old that influence the success of obtaining sufficient sweat for diagnosis of CF, and corresponding sweat chloride concentrations. A retrospective chart review of 119 sweat tests was performed on 103 preterm and full-term infants < or = 6 weeks of age. Bivariate and multivariate regression analyses were used to determine the predictors of successful sweat testing and characteristics influencing sweat chloride concentrations. Adequate amounts of sweat (> or = 75 mg) were obtained for analysis in 73.8% of initial attempts in the infant group. The following characteristics were associated with increased odds of obtaining a quantity not sufficient (QNS) for sweat chloride concentration measurement: African-American race, infant weight < 2,000 g, preterm birth, and postmenstrual age (PMA) < 36 weeks. With a multivariable logistic model, the only significant predictors were African-American race (7.3, 2.4-21.7) and PMA < 36 weeks (17.9, 4.2-75.9). Sweat chloride concentration in non-CF individuals is inversely related to both gestational age and age at testing, and this effect is additive in a linear regression model. In conclusion, sweat collection can be reliably performed in infants > or = 36 weeks postmenstrual age, > 2,000 g, and > 3 days postnatal age. Maturational factors have a mild impact on sweat chloride concentration. PMID- 15880421 TI - High prevalence of gastroesophageal reflux in children after lung transplantation. AB - Bronchiolitis obliterans and its clinical correlate bronchiolitis obliterans syndrome (BOS) are a major cause of morbidity and mortality following lung transplantation. Gastroesophageal reflux disease (GERD) may be a contributing factor for the development of BOS. Since 2002, all recipients of lung and heart lung transplantation at our institution have been routinely investigated for GERD. In this observational study, we report on the prevalence of GERD in this population, including all pediatric patients undergoing single (SLTx) or double (DLTx) lung transplantation or heart-lung (HLTx) transplantation from January 2003-May 2004. GERD was assessed 3-6 months after transplantation by 24-hr pH testing. The fraction time (Ft) with a pH < 4 within a 24-hr period was recorded. Spirometry data, episodes of confirmed acute rejection, and demographic data were also collected. Ten transplant operations were performed: 4 DLTx, 1 SLTx, and 5 HLTx. Nine patients had cystic fibrosis. One patient had end-stage pulmonary disease secondary to chronic aspiration pneumonia and postadenovirus lung damage. Of 10 patients tested, 2 had severe GERD (Ft > 20%), 5 had moderate GERD (Ft 10 20%), 2 had mild GERD (Ft 5-10%), and 1 had no GERD. The only patient in this group with no GERD had a Nissen fundoplication pretransplant. All study patients were asymptomatic for GERD. All patients with episodes of rejection had moderate to severe GERD posttransplant. There was no association between severity of GERD and peak spirometry results posttransplant. Moderate to severe GERD is common following lung transplantation in children. PMID- 15880422 TI - Light-dependent mutagenesis by benzo[a]pyrene is mediated via oxidative DNA damage. AB - Benzo[a]pyrene (B[a]P) is an environmental carcinogenic polycyclic aromatic hydrocarbon (PAH). Mammalian enzymes such as cytochrome P-450s and epoxide hydrase convert B[a]P to reactive metabolites that can covalently bind to DNA. However, some carcinogenic compounds that normally require metabolic activation can also be directly photoactivated to mutagens. To examine whether B[a]P is directly mutagenic in the presence of light, we exposed Salmonella typhimurium strains with different DNA repair capacities to B[a]P and white fluorescent light at wavelengths of 370-750 nm. B[a]P plus light significantly enhanced the number of His+ revertants. Mutagenesis was completely light-dependent and required no exogenous metabolic activation. The order of mutability of strains with different DNA repair capacities was strain YG3001 (uvrB, mutMST) >> strain TA1535 (uvrB) > strain YG3002 (mutMST) > strain TA1975. The uvrB gene product is involved in the excision repair of bulky DNA adducts, and the mutMST gene encodes 8-oxoguanine (8 oxoG) DNA glycosylase, which removes 8-oxoG from DNA. Introduction of a plasmid carrying the mOgg1 gene that is the mouse counterpart of mutMST substantially reduced the light-mediated mutagenicity of B[a]P in strain YG3001. B[a]P plus light induced predominantly G:C --> T:A and G:C --> C:G transversions. We propose that B[a]P can directly induce bulky DNA adducts if light is present, and that the DNA adducts induce oxidative DNA damage, such as 8-oxoG, when exposed to light. These findings have implications for the photocarcinogenicity of PAHs. PMID- 15880423 TI - Flow cytometric analysis of micronucleated reticulocytes: Time- and dose dependent response of known mutagens in mice, using multiple blood sampling. AB - According to the current Organization of Economic Cooperation and Development (OECD) and International Committee on Harmonization (ICH) guidelines for the mammalian erythrocyte micronucleus (MN) test, analysis of peripheral blood reticulocytes (RETs) for the presence of micronuclei can be performed using flow cytometry. The MicroFlow PLUS method (Litron Laboratories, Rochester, NY) for MN analysis by flow cytometry is based on the binding of FITC-labeled antibodies to the CD71 transferrin receptor of immature RETs, on parallel RNA degradation, and on propidium iodide staining of DNA present as micronuclei. The objective of this study was to assess the sensitivity of this flow cytometry method to detect time- and dose-dependent induction of micronuclei in mouse peripheral blood RETs after treatment with nine chemical agents. Five known clastogens, two known aneugens, and two compounds previously reported to be inactive in the mouse bone marrow MN test were evaluated at three dose levels. Multiple blood sampling of the same animal before and at two time points after treatment was conducted. All known mutagens produced a dose-dependent increase in micronucleated reticulocytes (MN RETs); the compounds previously shown to be inactive in the in vivo MN test were also negative using the present methodology. The highest frequency of MN-RETs was observed at 48 hr after treatment, except for 5-fluorouracil, which had its peak response at 72 hr. The results indicate that micronuclei can be measured by multiple blood sampling of the same animal before and after treatment without altering the sensitivity of the assay. The results confirm that the flow cytometric assessment of MN-RETs in mouse peripheral blood using MicroFlow PLUS is a sensitive method with high analysis throughput, and robust quality control. PMID- 15880424 TI - Retinitis following varicella in a vaccinated child with acute lymphoblastic leukemia. AB - Serious ocular disease following varicella (chickenpox) is rare in children. In addition, retinitis in children with hematologic malignancies may present a difficult diagnostic challenge because infectious retinitis may mimic leukemic involvement of the eye. We report a 7-year-old patient with T-cell acute lymphoblastic leukemia in remission who presented with visual complaints 2 weeks after developing chickenpox. Ophthalmologic evaluation revealed acute retinitis in the right eye. Prolonged therapy with acyclovir resulted in near complete recovery. Early diagnosis of VZV retinopathy and aggressive antiviral treatment is critical to prevent acute and long-term ocular sequelae. PMID- 15880425 TI - Identification of high frequency of Y chromosome deletions in patients with sex chromosome mosaicism and correlation with the clinical phenotype and Y-chromosome instability. AB - A mosaic karyotype consisting of a 45,X cell line and a second cell line containing a normal or an abnormal Y chromosome is relatively common and is associated with a wide spectrum of clinical phenotypes. The aim of this study was to investigate patients with such a mosaic karyotype for Y chromosome material loss and then study the possible association of the absence of these regions with the phenotype, diagnosis, and Y-chromosome instability. We studied 17 clinically well-characterized mosaic patients whose karyotype consisted of a 45,X cell line and a second cell line containing a normal or an abnormal Y chromosome. The presence of the Y chromosome centromere was verified by fluorescence in situ hybridization (FISH) and was then characterized by 44 Y-chromosome specific sequence tagged site (STS) markers. This study identifies a high frequency of Yq chromosome deletions (47%). The deletions extend from interval 5 to 7 sharing a common deleted interval (6F), which overlaps with the azoospermia factor region (AZF) region. This study finds no association between Y-chromosome loci hosting genes other than SRY, and the phenotypic sex, the diagnosis, and the phenotype of the patients. Furthermore, this study shows a possible association of these deletions with Y-chromosome instability. PMID- 15880426 TI - Th1 cytokine-induced downregulation of PPARgamma in human biliary cells relates to cholangitis in primary biliary cirrhosis. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) is known to inhibit the production of proinflammatory cytokines. In Th1-predominant diseases, PPARgamma ligands can ameliorate clinical severity by downregulating the expression of proinflammatory cytokines. Primary biliary cirrhosis (PBC) is characterized by chronic destructive cholangitis with a Th1-predominant cytokine milieu. Unusual immune responses to infectious agents are suspected to underlie its etiopathogenesis. We examined the significance of PPARgamma in biliary inflammation in connection to PBC. To this end, we performed immunohistochemistry, quantitative polymerase chain reaction, and nuclear factor kappaB (NF-kappaB) DNA-binding assays to clarify the intrahepatic distribution of PPARgamma and the regulation of PPARgamma by inflammatory cytokines and PPARgamma ligand in five cultured biliary cell lines including one derived from PBC liver. In liver specimens from patients with PBC, PPARgamma protein was ubiquitously expressed in intrahepatic biliary epithelium, whereas the expression of PPARgamma protein and mRNA was reduced in damaged bile ducts. PPARgamma expression in cultured cells was upregulated by interleukin-4 (IL-4; Th2-type), but downregulated by IFN-gamma (Th1-type). PPARgamma ligand negatively modulated lipopolysaccharide-induced NF-kappaB activation. Moreover, this inhibitory effect of PPARgamma ligand was attenuated by pretreatment with IFN-gamma. In conclusion, PPARgamma may be important to maintain homeostasis in the intrahepatic biliary epithelium, and its reduction in the bile ducts of PBC liver may be associated with the Th1-predominant milieu and with the development of chronic cholangitis in PBC. Immunosuppression using PPARgamma ligands may be of therapeutic benefit to attenuate biliary inflammation in PBC. PMID- 15880427 TI - Calcitonin gene polymorphism CALCA-624 (T/C) and ovarian cancer. AB - In a previous analysis, we reported an inverse association of dietary calcium intake with the risk of ovarian cancer (Goodman et al. 2002. Am J Epidemiol 156:148-57). The CALCA gene codes for calcitonin, an important regulator of bone calcium metabolism. Data from a population-based case-control study conducted in Hawaii were used to examine the hypothesis that a T --> C transition 624 base pairs upstream (-624) of the translation initiation codon of the CALCA gene influences the risk of ovarian malignancy. A structured interview was conducted for 182 histologically confirmed ovarian cancer cases and 219 controls. Blood specimens were collected from the subjects at their homes. A significant negative trend (P for trend: 0.02) in the odds ratios (ORs) was found with increasing intake of calcium. Women with any CALCA C allele were at nonsignificantly higher risk of ovarian cancer (OR: 1.5, 95% CI: 0.9-2.3) compared to women with the TT genotype and the risk increased with the number of C alleles (P for trend: 0.05). When further analyzed within ethnic subgroups, a significant positive association was found among Japanese for CALCA CT (OR: 2.3, 95% CI: 1.0-5.3) and CALCA CC (OR: 7.2, 95% CI: 1.1-46.0) compared with Japanese women who were homozygous for the T allele. The trend in risk associated with the C allele was most significant among women who had used oral contraceptives (P for trend: 0.05), had been pregnant (P for trend: 0.04), and had nonmucinous histological types of ovarian cancer (P for trend: 0.02). However, the association of ovarian cancer risk with the CALCA genotype was not significantly modified by any of the dietary, nondietary, or clinical variables included in this study. These preliminary data suggest a strong positive association of the CALCA C allele with the risk of ovarian cancer among some subgroups. PMID- 15880428 TI - Pathways to schizophrenic psychosis: a LISREL-tested model of the unfolding of the schizophrenic prodrome. AB - In this article a literature-based model (the Schizotypic Syndrome Questionnaire [SSQ] model) is presented that gives a description of the temporal unfolding of the schizophrenic prodrome. As a guiding principle for the selection of the symptoms in the model, the hypothesis was held that the main prodromal features determine each other in terms of cause and effect. Furthermore, the developmental pathways between the symptoms were not allowed to be in conflict with the usual observation that negative symptoms precede psychotic-like ones nor--at least in broad outline--with J.P. Docherty, D.P. van Kammen, S.G. Siris, and S.R. Marder's (1978) description of the various onset stages in the development of a schizophrenic psychosis. For the definitive version of the SSQ model, 12 symptoms were selected (e.g., affective flattening, suspicion, and delusional thinking). After specifying the paths to be estimated, the model was examined in two randomly drawn samples from a total community-based sample of 771 normal subjects and in the total sample itself, in each case resulting in adequate fit values. Moreover, all postulated pathways were found to be significantly different from zero. The use of a normal sample was based on the continuum hypothesis. Given the present-day discussions concerning the tenability of the schizophrenia concept, the model's implications with respect to that issue are particularly emphasized. Furthermore, the concept of the schizophrenia prodrome itself is critically discussed. PMID- 15880429 TI - Novel CD47-dependent intercellular adhesion modulates cell migration. AB - CD47 is a ubiquitously expressed plasma membrane protein, also known as Integrin Associated Protein, that modulates cell adhesion both through alteration of the avidity of integrin binding and through interaction with its own ligands, the extracellular matrix protein thrombospondin (TSP) and the plasma membrane response regulator SIRPalpha1. We now show that CD47 expression on fibroblasts can induce intercellular adhesion resulting in cell aggregation in the absence of active integrins, SIRPalpha1 binding, and detectable TSP. CD47-expressing cells preferentially bind to other CD47-expressing cells, and intercellular adhesion requires stimulation by serum or a CD47-binding peptide from TSP. Cell-cell adhesion is inhibited by pertussis toxin and C. difficile toxin B, and both adherent and aggregating CD47-expressing fibroblasts have more rac in the GTP bound state than CD47-deficient cells. Spontaneous migration of Jurkat lymphocytes through a fibroblast monolayer is decreased by fibroblast expression of CD47, consistent with an increased barrier function of the CD47 expressing cells. The lymphocyte chemoattractant SDF-1alpha stimulates migration of Jurkat cells through this monolayer only if both the lymphocytes and fibroblasts express CD47, and the inhibition of migration by a CD47-interacting peptide from TSP similarly requires CD47 expression on both cell types. Thus, signaling dependent on both heterotrimeric and rho family GTPases can induce CD47 to participate in cell-cell interactions independent of known ligands that enhance intercellular adhesion and modulate cell migration. PMID- 15880430 TI - Overexpression of neuronal intermediate filament protein alpha-internexin in PC12 cells. AB - The neuronal intermediate filaments include not only the neurofilament triplet proteins but also peripherin and alpha-internexin. To determine whether neurite outgrowth is enhanced by alpha-internexin, the cDNA of rat alpha-internexin tagged with enhanced green fluorescent protein (EGFP) was transfected into a rat adrenal pheochromocytoma cell line PC12 that responds to nerve growth factor (NGF) by induction of the neuronal phenotype. Selected stable clones were induced by NGF and examined for expression patterns of neuronal intermediate filaments by Western blot and immunocytochemistry. Differentiating neurons were also collected after NGF induction for RT-PCR analysis. Overexpressed alpha-internexin-EGFPs were found mainly in cell bodies and the proximal part of neurites. It was also found that overexpression of alpha-internexin-EGFPs enhanced the neurite outgrowth of PC12 cells at the early stages of NGF induction. Meantime, NF-L and NF-M were upregulated by the overexpression of alpha-internexin-EGFPs. Interestingly, alpha-internexin-EGFP-transfected cells obviously detached from culture plates at the later stages of NGF induction. Massive IF accumulations, swelling mitochondria, and degenerating neurites with numerous electron-dense granules were observed ultrastructurally in the alpha-internexin-EGFP-transfected cells. In addition, neuronal death was also characterized positively by the TUNEL assay. These observations may imply that cell death was occurring in alpha internexin-EGFP-transfected cells. From this study, it could be suggested that alpha-internexin plays an important role in neurite outgrowth and regulates the expression of other neurofilaments during neuronal development. Apoptosis-like cell death could also be induced by the overexpression of alpha-internexin-EGFP in PC12 cells after NGF induction. PMID- 15880431 TI - Reduced expression of the PTH/PTHrP receptor during development of the mammary gland influences the function of the nipple during lactation. AB - Signaling by the parathyroid hormone/parathyroid hormone-related protein receptor (Ppr) is necessary for mammary gland development beyond the early induction stage in mice. We used a series of murine models of reduced Ppr expression to determine how diminished receptor signaling influences mammary development. Reduction of Ppr expression to very low levels prevented mammary gland development. A less severe reduction in Ppr expression permitted progression of mammary gland development beyond the induction stage, but the nipples of these mice were dramatically smaller than those of controls, with altered epidermis and connective tissue. Mothers with reduced expression of Ppr could not successfully nurse pups; however, the lactating glands did produce milk but could not efficiently deliver it. This finding was associated with reduced levels of matrix metalloproteinase-2 and an absence of pregnancy-associated remodeling of connective tissue matrix in the nipple. Reduced smooth muscle appears to underlie the majority of nipple deficiencies in mice with lower levels of the Ppr expression. PMID- 15880432 TI - Recurrent central neurocytomas. AB - BACKGROUND: Since the first description of Central neurocytomas (CNs) as a benign tumor entity in 1982, there has been great enthusiasm regarding the benign course and the curative surgical approach to this disease. The current study was performed to investigate the frequency of disease recurrence during long-term follow-up. METHODS: A retrospective analysis of the medical files with emphasis on clinicoradiologic findings and histologic and immunohistochemical features was performed. RESULTS: Between 1985-2003. surgical resection was performed in 14 patients with CNs ages 16-43 years (7 were female and 7 were male). Two patients (14%) died postoperatively and one patient had a malignant disease course (7%). In the remaining 11 patients, one patient with an incompletely resected CN had disease progression after 37 months but at the time of last follow-up had had stable disease for 10 years. In addition, the authors reported 5 patients with disease recurrence occurring at a median of 67 months after surgery (range, 51-79 months after surgery), all of which occurred after complete surgical resection was performed. The observation period for the remaining 5 patients was short (median of 34 months [range, 5-44 months]). Extensive histologic and immunohistochemical workup did not identify any significant prognostic parameters. The MIB-1 proliferation index ranged from 0.8-11% (median of 4.6%), but was reported to be 46.8% in the malignant transformed tumor. All patients with disease recurrence responded well to different forms of focal radiation therapy (gamma knife radiosurgery in three patients and interstitial irradiation in one patient) and for one patient with a recently detected recurrence, gamma knife radiosurgery was planned. CONCLUSIONS: CNs appear to have a higher tendency to recur during long-term follow-up than previously reported, even after complete resection. Therefore, periodic neuroradiologic follow-up examinations should be considered mandatory in all patients, even after several years. PMID- 15880433 TI - Risk factors for infections with multidrug-resistant Pseudomonas aeruginosa in patients with cancer. AB - BACKGROUND: Pseudomonas aeruginosa is responsible for a wide range of infections. In immunocompromised patients with cancer, the emergence of multidrug resistant P. aeruginosa may have grave consequences. METHODS: Patients with cancer who were infected with multidrug-resistant P. aeruginosa with polyclonal DNA restriction patterns were used as the case group. Two control groups were used: one group of cancer patients who were infected with multidrug-susceptible P. aeruginosa and another group of cancer patients who had the same underlying disease and the same intensive care unit exposure as patients in the case group but who were not infected or colonized by P. aeruginosa. RESULTS: Risk factors that were associated significantly with multidrug-resistant P. aeruginosa infection were the use of carbapenem for > or = 7 days, a history of P. aeruginosa infection during the preceding year, and a history of chronic obstructive pulmonary disease (P < 0.01). CONCLUSIONS: Carbapenems may need to be used more judiciously as first-line empirical therapy for cancer patients with prior pseudomonal infection or chronic obstructive pulmonary disease who require hospitalization, and alternative, antipseudomonal antibiotic regimens may need to be considered, especially in this patient population. PMID- 15880434 TI - Comparative functional analysis of the hyolingual anatomy in lacertid lizards. AB - The tongue is often considered a key innovation in the evolution of a terrestrial lifestyle as it allows animals to transport food items through the oral cavity in air, a medium with low density and viscosity. The tongue has been secondarily coopted for a wide diversity of functions, including prey capture, drinking, breathing, and defensive behaviors. Within basal lizard groups, the tongue is used primarily for the purpose of prey capture and transport. In more derived groups, however, the tongue appears specialized for chemoreceptive purposes. Here we examine the tongue structure and morphology in lacertid lizards, a group of lizards where the tongue is critical to both food transport and chemoreception. Because of the different mechanical demands imposed by these different functions, regional morphological specializations of the tongue are expected. All species of lacertid lizards examined here have relatively light tongue muscles, but a well developed hyobranchial musculature that may assist during food transport. The intrinsic musculature, including verticalis, transversalis, and longitudinalis groups, is well developed and may cause the tongue elongation and retraction observed during chemoreception and drinking. The papillary morphology is complex and shows clear differences between the tongue tips and anterior fore-tongue, and the more posterior parts of the tongue. Our data show a subdivision between the fore- and hind-tongue in both papillary structure and muscular anatomy likely allowing these animals to use their tongues effectively during both chemoreception and prey transport. Moreover, our data suggest the importance of hyobranchium movements during prey transport in lacertid lizards. PMID- 15880435 TI - Geographic and temporal variations in cancer of the corpus uteri: incidence and mortality in pre- and postmenopausal women in Europe. AB - Corpus uteri cancer is the fourth most common neoplasm in women in Europe and the tenth most common cause of cancer death. We examined geographic and temporal variations in corpus uteri cancer incidence and mortality rates in the age groups 25-49 and 50-74 in 22 European countries. The disease is considerably less common in premenopausal women, with incidence and mortality rates decreasing throughout Europe and mortality declines more marked in western and southern European countries. Incidence rates among postmenopausal women are highest in the Czech Republic, Slovakia, Sweden and Slovenia and lowest in France and the United Kingdom. Increasing incidence trends in this age group are observed in the Nordic countries (except Denmark) and in the United Kingdom. Some increases are also seen in eastern (Slovakia) and southern Europe (Spain and Slovenia), while relatively stable or modestly decreasing trends are observed in Italy and most western European countries. Postmenopausal mortality rates are systematically higher in eastern Europe, with death rates in the Ukraine, Latvia, Czech Republic, Russia and Belarus 2-3 times those seen in western Europe. Declining mortality trends are seen in most populations, though in certain Eastern European countries, the declines began rather recently, during the 1980s. In Belarus and Russia, recent postmenopausal death rates are stable or increasing. The rates are adjusted for misclassification of uterine cancer deaths but remain unadjusted for hysterectomy, and where there is an apparent levelling off of incidence or mortality rates recently, rising prevalence of hysterectomy cannot be discounted as an explanation. However, the trends by age group can be viewed in light of several established risk factors for endometrial cancer that are highly prevalent and most likely changing with time. These are discussed, as are the prospects for preventing the disease. PMID- 15880436 TI - Clinical psychologists across the years: the division of clinical psychology from 1960 to 2003. AB - For more than 40 years researchers have studied the members of the American Psychological Association's (APA) Division of Clinical Psychology to obtain information about their demographic characteristics, educational experiences, theoretical orientations, employment settings, professional activities, publication histories, and career satisfactions. We summarize the results from the most recent study (N=694, 46% return rate) in the historical context of the previous findings, dating back to 1960. We provide both contemporary and historical portraits of American clinical psychology. Among the most notable trends are a steady increase in female psychologists, a decline in psychological assessment in general and projective testing in particular, the modal eclectic/integrative orientation being rivaled by the cognitive orientation, a pattern of high career satisfaction, and continued enthusiasm for the Boulder model. PMID- 15880437 TI - Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate. AB - The RNA helicase Vasa is a germ cell marker in animals, and its homolog in vertebrates to date has been limited to bisexual reproduction. We cloned and characterized CagVasa, a Vasa homolog from the gibel carp, a fish that reproduces bisexually or gynogenetically. CagVasa possesses 14 RGG repeats and eight conserved motifs of Vasa proteins. In bisexually reproducing gibel carp, vasa is maternally supplied and its zygotic expression is restricted to gonads. By in situ hybridization on testicular sections, vasa is low in spermatogonia, high in primary spermatocytes, reduced in secondary spermatocytes, but disappears in spermatids and sperm. In contrast, vasa persists throughout oogenesis, displaying low-high-low levels from oogonia over vitellogenic oocytes to maturing oocytes. A rabbit anti-Vasa antibody (alphaVasa) was raised against the N-terminal CagVasa for fluorescent immunohistochemistry. On testicular sections, Vasa is the highest in spermatogonia, reduced in spermatocytes, low in spermatids, and absent in sperm. In the ovary, Vasa is the highest in oogonia but persists throughout oogenesis. Subcellular localization of vasa and its protein changes dynamically during oogenesis. The alphaVasa stains putative primordial germ cells in gibel carp fry. It detects gonadal germ cells also in several other teleosts. Therefore, Cagvasa encodes a Vasa ortholog that is differentially expressed in the testis and ovary. Interestingly, the alphaVasa in combination with a nuclear dye can differentiate critical stages of spermatogenesis and oogenesis in fish. The cross-reactivity and the ability to stain stage-specific germ cells make this antibody a useful tool to identify fish germ cell development and differentiation. PMID- 15880438 TI - Disruption of Sertoli-germ cell adhesion function in the seminiferous epithelium of the rat testis can be limited to adherens junctions without affecting the blood-testis barrier integrity: an in vivo study using an androgen suppression model. AB - During spermatogenesis, both adherens junctions (AJ) (such as ectoplasmic specialization (ES), a testis-specific AJ type at the Sertoli cell-spermatid interface (apical ES) or Sertoli-Sertoli cell interface (basal ES) in the apical compartment and BTB, respectively) and tight junctions (TJ) undergo extensive restructuring to permit germ cells to move across the blood-testis barrier (BTB) as well as the seminiferous epithelium from the basal compartment to the luminal edge to permit fully developed spermatids (spermatozoa) to be sloughed at spermiation. However, the integrity of the BTB cannot be compromised throughout spermatogenesis so that postmeiotic germ cell-specific antigens can be sequestered from the systemic circulation at all times. We thus hypothesize that AJ disruption in the seminiferous epithelium unlike other epithelia, can occur without compromising the BTB-barrier, even though these junctions, namely TJ and basal ES, co-exist side-by-side in the BTB. Using an intratesticular androgen suppression-induced germ cell loss model, we have shown that the disruption of AJs indeed was limited to the Sertoli-germ cell interface without perturbing the BTB. The testis apparently is using a unique physiological mechanism to induce the production of both TJ- and AJ-integral membrane proteins and their associated adaptors to maintain BTB integrity yet permitting a transient loss of cell adhesion function by dissociating N-cadherin from beta-catenin at the apical and basal ES. The enhanced production of TJ proteins, such as occludin and ZO-1, at the BTB site can supersede the transient loss of cadherin-catenin function at the basal ES. This thus allows germ cell depletion from the epithelium without compromising BTB integrity. It is plausible that the testis is using this novel mechanism to facilitate the movement of preleptotene and leptotene spermatocytes across the BTB at late stage VIII through early stage IX of the epithelial cycle in the rat while maintaining the BTB immunological barrier function. PMID- 15880439 TI - Ubiquitous expression of mRFP1 in transgenic mice. AB - Fluorescent proteins provide a powerful means to track gene expression and cellular behaviors in the study of model organisms such as mice. Among the new generation of fluorescent protein markers, the monomeric red fluorescent protein mRFP1 is particularly attractive because of its rapid maturation and minimal interference with GFP and GFP-derived markers. Here we evaluate the utility of mRFP1 as a marker in transgenic mice. We show that high level and ubiquitous expression of mRFP1 does not affect mouse development, general physiology, or reproduction. mRFP1 expression can be readily detected with unaided eyes under daylight in transgenic mice on the albino background. The intensity of mRFP1 signals can be used to distinguish homozygous and heterozygous transgenic mice. Together, these features make mRFP1 an attractive marker for broad applications in transgenic research. PMID- 15880440 TI - Gicerin/CD146 is involved in neurite extension of NGF-treated PC12 cells. AB - Gicerin/CD146 is a cell adhesion molecule, which belongs to the immunoglobulin (Ig) superfamily. We have reported that it has a homophilic binding activity, which participates in the neurite extension from embryonic neurons. To elucidate how gicerin is involved in the neurite extension mechanism, we employed PC12 cells, which expresses gicerin/CD146. PC12 cells extend longer neurites by nerve growth factor (NGF) on gicerin substrate than on without gicerin substrate, which indicates that gicerin participates in neurite extension by NGF. We also found that the expression of gicerin in PC12 cells is induced by NGF. Over-expression of gicerin also promotes neurite extension by gicerin-gicerin homophilic interaction. These findings suggested that increase of gicerin expression by NGF promotes the gicerin-gicerin homophilic interaction resulting in the neurite extension. PMID- 15880441 TI - Utilization of fluoroquinolones and Escherichia coli resistance in urinary tract infection: inpatients and outpatients. AB - OBJECTIVES: The aim of this study was to evaluate the dependence of Escherichia coli resistance to fluoroquinolones on their use in the outpatients and inpatients in the Olomouc region of the Czech Republic. METHODS: Data on inpatient antibiotic use were obtained from the database of the Department of Pharmacology and expressed as defined daily dose per 100 bed-days (DBD). Data on outpatient prescriptions were obtained from the database of General Health Insurance Company and expressed in defined daily doses per 1000 clients per day (DID). Escherichia coli strains were isolated from samples of urine of both community and hospitalized patients suffering from acute bacterial urinary tract infection, examined using aerobic cultivation, and determined by standard biochemical procedures. RESULTS: The utilization of fluoroquinolones in inpatients has significantly (p < 0.01) increased from 2.52 DBD in 1997 to 4.29 DBD in 2002. In outpatients, fluoroquinolone utilization has also increased significantly from 0.14 to 0.95 DID (p < 0.01). In the same period, 9192 E. coli strains were isolated from inpatients and outpatients suffering by urinary tract infections and tested for the susceptibility to fluoroquinolones. Resistance increased significantly (p < 0.01) both in the hospital (from 2 to 9%) and in the community (from 1 to 10%). The development of E. coli resistance to fluoroquinolones correlates significantly with their utilization both in the hospital (r = 0.944, p = 0.005) and in the community (r = 0.859, p = 0.029). CONCLUSIONS: Results of this study shows the impact of fluoroquinolone utilization on E. coli resistance and support the need of controlled use of these effective antibiotics. PMID- 15880442 TI - Contact allergy to neomycin sulfate: results of a multifactorial analysis. AB - PURPOSE: To perform a comprehensive, multifactorial analysis of potential risk factors (demographic and clinical) for contact allergy to neomycin sulfate, a common adverse reaction resulting from the topical use of this drug; especially in some subgroups of the population. METHODS: Retrospective analysis of allergy test data of the Information Network of Departments of Dermatology (IVDK, www.ivdk.org) between 1998 and 2003, including all patients patch tested with a standard screening series because of suspected allergic contact dermatitis (ACD). As one outcome, a positive (allergic) test reaction to neomycin sulfate was considered. An alternative outcome included only those patients with a positive test to neomycin sulfate and a final diagnosis of ACD. The association between outcome and potential risk factors was analyzed with Poisson regression analysis, deriving prevalence ratios (PR) as risk estimates. RESULTS: Of the 47,559 patients tested, 2.5% had positive reactions to neomycin sulfate, while in 1.1% ACD was additionally diagnosed. The results of the multifactorial analysis indicated that the risk of both outcomes decreased slightly during the period covered; was higher among patients with leg dermatitis; varied significantly with age and increased progressively with the number of additional positive reactions to other standard series allergens. Cross-reactivity to other, selectively tested, aminoglycoside antibiotics was substantial (kappa = 0.67; 95%CI: 0.63 0.71) for framycetin sulfate, to low (kappa = 0.33; 95%CI: 0.27-0.37) for gentamicin sulfate. CONCLUSIONS: The prevalence of contact sensitization to neomycin sulfate was noteworthy among patients patch tested in the IVDK centers. Supplementing clinical epidemiology, neomycin contact allergy has been estimated to be relatively common even on the level of the unselected population (prevalence approx. 1%). Hence, the topical use of neomycin sulfate by patients should be carefully monitored, considering its potential to induce ACD, with emphasis on subgroups at risk. PMID- 15880443 TI - Syndromes and epistemology I: Autistic spectrum disorders. PMID- 15880444 TI - Inhibition of colon cancer cell proliferation by the dietary compound conjugated linoleic acid is mediated by the CDK inhibitor p21CIP1/WAF1. AB - Our previous studies indicated that dietary conjugated linoleic acid (CLA) inhibits colon tumor cell proliferation in vitro and in vivo. To identify mechanisms by which CLA regulates growth arrest, the HT-29 human colon carcinoma cell line was treated with various physiological concentrations of CLA and analyzed by flow cytometry. We detected a dose-dependent increase in the percentage of cells arrested in G1 after CLA treatment that was accompanied by induction of the cyclin dependent kinase (CDK) inhibitor p21CIP1/WAF. CLA addition also led to increased p21 expression in HCT116 and SW480 cells, indicating that p21 induction is a general consequence of CLA treatment in colon cancer cells. Since both HT-29 and SW480 cells have mutant p53, our data indicate that p53 is not essential for induction of p21. In addition to an increase in p21 levels, HT-29 cell growth arrest was also accompanied by moderate decreases in Cyclin A, D1, E, and proliferating cell nuclear antigen (PCNA) levels. Following CLA treatment, p21 associated with and inhibited CDK4 and CDK2, and this correlated with reduced phosphorylation of retinoblastoma proteins. Increased association of p21 with PCNA was also detected. Dietary CLA inhibits cell cycle progression by inducing p21, which negatively regulates the growth promoting activities of CDK/cyclins and PCNA. These studies indicate that physiological concentrations of CLA inhibit growth of colon cancer cells with either wild-type or mutant p53, and may have therapeutic benefits in vivo. PMID- 15880445 TI - Effect of adenosine triphosphate on phosphate uptake in renal proximal tubule cells: involvement of PKC and p38 MAPK. AB - ATP has been known to act as an extracellular signal and to be involved in various functions of kidney. Renal proximal tubular reabsorption of phosphate (Pi) contributes to the maintenance of phosphate homeostasis, which is regulated by Na+/Pi cotransporter. However, the effects of ATP on Na+/Pi cotransporters were not elucidated in proximal tubule cells (PTCs). Thus, the effects of ATP on Na+/Pi cotransporter and its related signal pathways are examined in the primary cultured renal PTCs. In the present study, ATP inhibited Pi uptake in a time (> 1 h) and dose (>10(-6)M) dependent manner. ATP-induced inhibition of Pi uptake was correlated with the decrease of type II Na+/Pi cotransporter mRNA. ATP-induced inhibition of Pi uptake may be mediated by P2Y receptor activation, since suramin (non-specific P2 receptor antagonist) and RB-2 (P2Y receptor antagonist) blocked it. ATP-induced inhibition of Pi uptake was blocked by neomycin, U73122 (phospholipase C (PLC) inhibitors), bisindolylmaleimide I, H-7, and staurosporine (protein kinase C (PKC) inhibitors), suggesting the role of PLC/PKC pathway. ATP also increased inositol phosphates (IPs) formation and induced PKC translocation from cytosolic fraction to membrane fraction. In addition, ATP-induced inhibition of Pi uptake was blocked by SB 203580 [a p38 mitogen activated protein kinase (MAPK) inhibitor], but not by PD 98059 (a p44/42 MAPK inhibitor). Indeed, ATP induced phosphorylation of p38 MAPK, which was not blocked by PKC inhibitor. In conclusion, ATP inhibited Pi uptake via PLC/PKC as well as p38 MAPK in renal PTCs. PMID- 15880446 TI - Effect of selenium-supplement on the calcium signaling in human endothelial cells. AB - Intracellular Ca2+ signaling controls many cellular functions. Understanding its regulation by selenoproteins is essential for understanding the role of selenoproteins in regulating cell functions. The activity of thioredoxin reductase (TrxR), thioredoxin (Trx) content, and the activity of glutathione peroxidase (GPx) in the human endothelial cells cultured in selenium-supplemented medium (refer as Se+ cells) was found 70%, 40%, and 20% higher, respectively than those in the cells cultured in normal medium (refer as Se0 cells). The intracellular Ca2+ signaling initiated by inositol 1,4,5-trisphosphate (IP3), histamine, thapsigargin (TG), carbonyl cyanide p-(tri-fluoromethoxy) phenyl hydrazone (FCCP), and cyclosporin A (CsA) was investigated in both Se+ and Se0 cells. It was interestingly found that the higher activity of selenoproteins reduced the sensitivity of IP3 receptor to the IP3-triggered Ca2+ release from intracellular stores, but enhanced activation of the receptor-coupled phospholipase C in histamine-stimulated Se+ cells by showing much more generation of IP3 and higher elevation of cytosolic Ca2+. The higher selenoprotein activity also reduced susceptibility of the uniporter to the mitochondrial uncoupler, susceptibility of the permeability transition pore (PTP) to its inhibitor, and the vulnerability of endoplasmic reticulum (ER) Ca2+-ATPase to its inhibitor in selenium-supplementing cells. The results suggest that cell calcium signaling is subjected to thiol-redox regulation by selenoproteins. PMID- 15880447 TI - Store-operated Ca2+ entry: vesicle fusion or reversible trafficking and de novo conformational coupling? AB - Store-operated Ca2+ entry (SOCE), a mechanism regulated by the filling state of the intracellular Ca2+ stores, is a major pathway for Ca2+ influx. Hypotheses to explain the communication between the Ca2+ stores and plasma membrane (PM) have considered both the existence of small messenger molecules, such as a Ca2+-influx factor (CIF), and both stable and de novo conformational coupling between proteins in the Ca2+ store and PM. Alternatively, a secretion-like coupling model based on vesicle fusion and channel insertion in the PM has been proposed, which shares some properties with the de novo conformational coupling model, such as the role of the actin cytoskeleton and soluble N-ethylmaleimide (NEM)-sensitive factor attachment proteins receptor (SNARE) proteins. Here we review recent progress made in the characterization of the de novo conformational coupling and the secretion-like coupling models for SOCE. We pay particular attention into the involvement of SNARE proteins and the actin cytoskeleton in both SOCE models. SNAREs are recognized as proteins involved in exocytosis, participating in vesicle transport, membrane docking, and fusion. As with secretion, a role for the cortical actin network in Ca2+ entry has been demonstrated in a number of cell types. In resting cells, the cytoskeleton may prevent the interaction between the Ca2+ stores and the PM, or preventing fusion of vesicles containing Ca2+ channels with the PM. These are processes in which SNARE proteins might play a crucial role upon cell activation by directing a precise interaction between the membrane of the transported organelle and the PM. PMID- 15880448 TI - Gemtuzumab ozogamicin exposure and portal fibrosis. PMID- 15880449 TI - Acute ischemic cardiac dysfunction is attenuated via gene transfer of a peptide inhibitor of the beta-adrenergic receptor kinase (betaARK1). AB - Acute myocardial ischemia is a critical adverse effect potentially occurring during cardiac procedures. A peptide inhibitor of the beta-adrenergic receptor kinase (betaARK1), betaARKct, has been successful in rescuing chronic myocardial ischemia. The present study focused on the effects of adenoviral-mediated betaARKct (Adv-betaARKct) delivery on left ventricle (LV) dysfunction induced by acute coronary occlusion. Rabbits received intracoronary delivery of phosphate buffered saline (PBS) (n=9) or 5x10(11) viral particles of betaARKct (n=8). A loose prolene 5-0 Potz-loop suture was placed around the circumflex coronary artery (LCx) with both ends buried under the skin. Four days later, the suture was retrieved and pulled to occlude the LCx. Ischemia was confirmed by immediate ECG changes. LV function was continuously recorded for 45 min. Contractility (LVdP/dtmax), relaxation (LVdP/dtmin) and end diastolic pressure (EDP) were less impaired in the betaARKct group as compared to PBS (P<0.05, two-way ANOVA). betaAR density was higher in the ischemic area of the LV in the betaARKct group (betaARKct: 71.9+/-4.6 fmol/mg protein, PBS: 54.5+/-4.0 fmol/mg protein, P<0.05). Adenylyl cyclase activity was also improved basally and in response to betaAR stimulation. betaARK1 activation was less in the betaARKct group (P<0.05). Therefore, inhibition of myocardial betaARK1 may represent a new strategy to prevent LV dysfunction induced by acute coronary ischemia. PMID- 15880450 TI - Preadipocytes in the human subcutaneous adipose tissue display distinct features from the adult mesenchymal and hematopoietic stem cells. AB - The stroma-vascular fraction (SVF) of human adipose tissue has recently been described to be composed of endothelial cells identified as CD34+/CD31+ cells, infiltrated/resident macrophages defined as CD14+/CD31+ cells, and a new cell population characterized as CD34+/CD31- cells. To elucidate the cell identity of the adipocyte precursor cells, fluorescent activating cell sorter (FACS) analyses were performed on crude SVF cultured under adipogenic conditions, i.e., serum deprived medium containing insulin, cortisol, triiodothyronine, and supplemented with a PPARgamma agonist for the first 3 days. The progressive accumulation of lipid droplets was associated with a selective enrichment of the CD34+/CD31- cell population whereas control experiments performed in medium supplemented with 10% serum showed an overall downregulation of the three cell markers without adipogenesis. Among the different cell subsets, the CD34+/CD31- subset was the unique cell fraction able to answer to adipogenic culture conditions. Indeed, a time-dependent expression of adipocyte markers as well as acquisition of adipocyte-typical metabolic activities were observed. In parallel, the gene expression of lipogenic and lipolytic enzymes increased. The ability to differentiate into adipocytes was restricted to cells that did not express the mesenchymal stem cell marker CD105. Furthermore, the CD34+/CD31- cells did not respond to culture conditions used for hematopoietic colony assays. Taken together, the present study demonstrates that adipocyte progenitor cells, i.e., the preadipocytes, are included in the CD34+/CD31- cell fraction, which displays distinct features from the adult mesenchymal and hematopoietic stem cells. PMID- 15880451 TI - Erythroid cell differentiation is characterized by nuclear matrix localization and phosphorylation of protein kinases C (PKC) alpha, delta, and zeta. AB - Protein kinases C (PKC) zeta expression and phosphorylation at nuclear level during dimethyl sulfoxide (DMSO)-induced differentiation in Friend erythroleukemia cells have been previously reported, suggesting a possible role of this PKC isoform in the DMSO-related signaling. In order to shed more light on this tantalizing topic, we investigated PKC intracellular and sub-cellular localization and activity during DMSO-induced erythroid differentiation. Results indicated that at least PKC alpha, zeta, and delta are strongly and temporally involved in the DMSO-induced differentiation signals since their expression and phosphorylation, though at different extents, were observed during treatments. Intriguingly, while PKC alpha and zeta associate to the nuclear matrix during the differentiation event, PKC delta appears to be residentially associated to the nuclear matrix. Furthermore, an evident downregulation of the beta-globin gene transcription (differentiation hallmark) was detected upon a progressive inhibition of these PKC isoforms by means of specific inhibitors, indicating, therefore, that PKC alpha, zeta, and delta phosphorylation play a crucial role in the control of erythroid differentiation. PMID- 15880452 TI - Structural basis for preferential binding of non-ortho-substituted polychlorinated biphenyls by the monoclonal antibody S2B1. AB - Polychlorinated biphenyls (PCBs) are a family of 209 isomers (congeners) with a wide range of toxic effects. In structural terms, they are of two types: those with and those without chlorines at the ortho positions (2, 2', 6 and 6'). Only 20 congeners have no ortho chlorines. Three of these are bound by the aryl hydrocarbon receptor and are one to four orders of magnitude more toxic than all others. A monoclonal antibody, S2B1, and its recombinant Fab have high selectivity and nanomolar binding affinities for two of the most toxic non-ortho chlorinated PCBs, 3,4,3',4'-tetrachlorobiphenyl and 3,4,3',4',5' pentachlorobiphenyl. To investigate the basis for these properties, we built a three-dimensional structure model of the S2B1 variable fragment (Fv) based on the high-resolution crystallographic structures of antibodies 48G7 and N1G9. Two plausible conformations for the complementarity-determining region (CDR) H3 loop led to two putative PCB-binding pockets with very different shapes (models A and B). Docking studies using molecular mechanics and potentials of mean force (PMF) indicated that model B was most consistent with the selectivity observed for S2B1 in competition ELISAs. The binding site in model B had a deep, narrow pocket between V(L) and V(H), with a slight constriction at the top that opened into a wider pocket between CDRs H1 and H3 on the antibody surface. This binding site resembles those of esterolytic antibodies that bind haptens with phenyl rings. One phenyl ring of the PCB fits into the deep pocket, and the other ring is bound in the shallower one. The bound PCB is surrounded by the side chains of TyrL91, TyrL96 and TrpH98, and it has a pi-cation interaction with ArgL46. The tight fit of the binding pocket around the ortho positions of the bound PCBs indicates that steric hindrance of ortho chlorines in the binding site, rather than induced conformational change of the PCBs, is responsible for the selectivity of S2B1. PMID- 15880454 TI - Protective effects of bone marrow stromal cell transplantation in injured rodent brain: synthesis of neurotrophic factors. AB - Several groups have suggested that transplantation of marrow stromal cells (MSCs) promotes functional recovery in animal models of brain trauma. Recent studies indicate that tissue replacement by this method may not be the main source of therapeutic benefit, as transplanted MSCs have only limited ability to replace injured central nervous system (CNS) tissue. To gain insight into the mechanisms responsible for such effects, we systematically investigated the therapeutic potential of MSCs for treatment of brain injury. Using in vitro studies, we detected the synthesis of various growth factors, including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), and neurotrophin-3 (NT-3). Enzyme-linked immunosorbent assay (ELISA) demonstrated that MSCs cultured in Dulbecco's modified Eagle medium (DMEM) produced substantial amounts of NGF for at least 7 weeks, whereas the levels of BDNF, GDNF and NT-3 remained unchanged. In studies in mice, after intraventricular injection of MSCs, NGF levels were increased significantly in cerebrospinal fluid by ELISA, confirming our cell culture results. Further studies showed that treatment of traumatic brain injury with MSCs could attenuate the loss of cholinergic neuronal immunostaining in the medial septum of mice. These studies demonstrate for the first time that by increasing the brain concentration of NGF, intraventricularly transplanted MSCs might play an important role in the treatment of traumatic brain injury. PMID- 15880455 TI - Thr 446 phosphorylation of PKR by HCV core protein deregulates G2/M phase in HCC cells. AB - Hepatitis C virus (HCV) is the major causative viral agent of cirrhosis and hepatocarcinoma (HCC). HCV core protein affects cell homeostasis, playing an important role in viral pathogenesis of HCC. We investigate the effects of HCV core protein expression on cell growth in HCC cell lines. Cell cycle distribution analysis of HepG2 polyclonal core positive cells reveals a peculiar accumulation of cells in G2/M phase. Different pathways mediate G2/M arrest: such as p53 and double strand RNA protein kinase (PKR). Flow cytometry in p53-null cells demonstrates that p53 plays only a marginal role in inducing HCV core-dependent G2/M phase accumulation that seems to be significantly affected by the functional inactivation of PKR. HCC core positive cells are characterized by a significant PKR phosphorylation in Thr 446 residue, which leads deregulation of mitosis. Moreover, we observe that the overexpression of the viral protein induces an upregulation of PKR activity, which does not correlate with an increased eIF-2 phosphorylation. This uncommon behavior of PKR suggests that its activation by HCV core protein could involve alternative PKR-dependent pathways, implicated in core-dependent G2/M accumulation. The described biological effects of HCV core protein on cell cycle could be an additional viral mechanism for both HCV resistance to interferon (IFN) and HCC HCV-related pathogenesis. PMID- 15880456 TI - Chondromyxoid fibroma resembles in vitro chondrogenesis, but differs in expression of signalling molecules. AB - Chondromyxoid fibroma is a rare benign cartilaginous bone tumour characterized by morphological features that resemble different steps of chondrogenesis in terms of both cellular morphology, ranging from spindled to rounded cells, and the extracellular matrix formed, which ranges from fibrous to cartilaginous. The presence in chondromyxoid fibroma of signalling molecules that regulate the spatial expression of proteins involved in normal cartilage proliferation and differentiation was investigated in samples from 20 patients and compared with articular chondrocytes from 11 normal donors cultivated in 3D pellet culture. Sections were stained with safranin-O and H&E, and immunohistochemistry was performed for p16, cyclin D1, FGFR3, BCL2, p21, PTHLH, PTHR1 and N-cadherin. Expression patterns were analysed using hierarchical clustering. In chondromyxoid fibroma, specific morphological features correlated with a distinct pattern of expression. Comparison with normal chondrocytes in pellet culture showed a striking morphological resemblance, but with an unmistakably different pattern of expression. N-cadherin, PTHLH, and PTHR1 were expressed to a significantly higher level (p < 0.01) in articular chondrocyte pellets but, conversely, there was significantly lower expression of cyclin D1, p16 and BCL2 (p < 0.05) in these cells. Morphological similarities reflect common steps in cartilage differentiation, albeit driven by different molecular mechanisms. The proteins we have found to be differentially expressed seem crucial for neoplastic chondrogenesis. PMID- 15880457 TI - Localization of a putative low-penetrance ependymoma susceptibility locus to 22q11 using a chromosome 22 tiling-path genomic microarray. AB - Ependymomas frequently display allelic loss of chromosome 22 in the absence of mutations in the known tumor-suppressor genes on chromosome 22, suggesting the role of an alternative predisposing gene or genes from this chromosome. In an effort to localize these genes, 37 ependymomas derived from 33 patients were analyzed for the presence of copy number changes by use of a high-resolution chromosome 22 genomic microarray. Eighteen ependymomas (49%) displayed an array CGH profile consistent with monosomy of chromosome 22. However, in 10 of these tumors, the fluorescence ratios for 22q clones scored as deleted were different from those at the single gene copy level. This suggests either analysis of mixed populations of tumor and normal stromal cells or analysis of mixed tumor cell populations with different genetic profiles. Four ependymomas derived from two patients showed overlapping interstitial deletions of 2.2 Mb and approximately 510 kb. Further analyses revealed that these deletions were present in the constitutional DNA of these two patients as well as in some of their unaffected relatives. Detailed microsatellite analysis of these families refined the commonly deleted segment to a region of 320 kb between markers RH13801 and D22S419. Our results provide additional evidence for the involvement of genes on chromosome 22 in the development of ependymoma and suggest the presence of a low penetrance ependymoma susceptibility locus at 22q11. PMID- 15880458 TI - Spirituality influences health related quality of life in men with prostate cancer. AB - Spirituality is interdependent with the biological, psychological, and interpersonal aspects of life. Although spirituality has been studied in breast cancer survivors, little work has been done in men with prostate cancer. We sought to determine whether lower spirituality in men with early stage prostate cancer is associated with worse general health-related quality of life (HRQOL), disease-specific HRQOL, or psychosocial health. Two hundred and twenty-two subjects were drawn from a state-funded program providing free prostate cancer treatment to indigent men. Validated instruments captured spirituality, general and disease-specific HRQOL, anxiety, symptom distress, and emotional well-being. We found a consistent relationship between spirituality and the outcomes assessed. Low spirituality was associated with significantly worse physical and mental health, sexual function and more urinary bother after controlling for covariates. All of the psychosocial variables studied reflected worse adjustment in the men with low spirituality. Because the likelihood of prostate cancer survivorship is high, interventions targeting spirituality could impact the physical and psychosocial health of many men. PMID- 15880459 TI - Postnatal ontogeny, population structure, and extinction of the giant moa Dinornis. AB - Recent reinterpretation of the giant moa Dinornis as consisting of two sexually dimorphic allospecies permits thorough site-by-site investigation of the ontogeny and population biology of this genus. Analysis of subadult skeletal material from natural swamp sites in the North and South Islands of New Zealand forms the basis for recognition of growth series for each long bone element, characterized by sequential formation of fossulae in the femur and fusion of bones in the tibiotarsus and tarsometatarsus. Femora reached progressive developmental stages more rapidly than the other long bones, but all three elements reached maturity at about the same time. Patterns of bone fusion in Dinornis are more similar to those in Apteryx than in Struthio, and kiwi are recognized as a suitable developmental analog for interpreting moa ontogeny. Samples from Bell Hill Vineyard Swamp (South Island) and Makirikiri swamp (North Island) are interpreted as representing autochthonous moa populations; comparison with stages of kiwi long bone development suggests that Dinornis at these sites had high adult survivorship in strongly K-selected populations, with 72.5-87.3% of individuals having achieved adult body mass, and 55.9-78.2% being sexually mature. The pattern of low fecundity and probable high longevity in both Dinornis species suggests that populations were vulnerable to loss of adults, primarily through hunting, rather than as a result of habitat destruction. PMID- 15880460 TI - Experimental analysis of the transdifferentiation of visceral to parietal endoderm in the mouse. AB - The visceral endoderm (VE) of isolated extraembryonic regions (ExEmbs) of 7 days postcoitum (dpc) prestreak mouse conceptuses have been shown to convert readily to parietal endoderm (PE). The present study addresses the following three unanswered questions. On what does conversion depend, how rapidly does it occur, and is it an enduring general property of a residual small population of relatively immature cells? In situ hybridization reveals that change in cell state occurs within 2 days of culture. Deprivation of the mesoderm also promotes it in later ExEmbs. Conversely, the conversion to PE in isolated 7 dpc ExEmbs is suppressed by grafting 8 dpc or 9 dpc mesoderm. Hence, the conversion provides an example of transdifferentiation that is promoted by the absence of extraembryonic mesoderm. The presence of mesoderm seems to be necessary to enable the VE to grow rather than convert to PE, as occurs if it retains contact with the extraembryonic ectoderm. PMID- 15880461 TI - Fluorescence resonance energy transfer analysis of apolipoprotein E C-terminal domain and amyloid beta peptide (1-42) interaction. AB - The potential neurotoxicity of soluble forms of amyloid beta peptide (Abeta) as a key factor in early pathogenesis of Alzheimer's disease is being recognized. In addition, there is growing evidence of the essential role of apolipoprotein E (apoE) in amyloid formation, although molecular details of apoE/Abeta interaction are poorly understood. We employed apoE C-terminal (CT) domain comprising residues 201-299 to identify binding location of Abeta(1-42) by fluorescence resonance energy transfer (FRET) and quenching analyses. Native tryptophan (Trp) residues in the apoE CT domain served as FRET donor, whereas N-(iodoacetyl)-N'-(5 sulfo-1-naphthyl)ethylenediamine (AEDANS) covalently attached to a unique cysteine residue substituted at position 4 of Abeta(1-42) (AEDANS-F4C-Abeta(1 42)) served as FRET acceptor. Fluorescence analysis verified that the oligomerization behavior of AEDANS-F4C-Abeta(1-42) was not abrogated by covalent attachment of AEDANS and that apoE CT domain/AEDANS-F4C-Abeta(1-42) association results in formation of a soluble complex. A large decrease in Trp fluorescence emission was noted in mixtures containing apoE CT domain and AEDANS-F4C-Abeta(1 42), accompanied by appearance of sensitized fluorescence emission of AEDANS as a result of intermolecular FRET. An average distance of separation of 22.6 Angstroms between donors and acceptor was calculated. Fluorescence quenching by potassium iodide (KI) did not reveal significant differences in apoE CT domain Trp microenvironment in the absence or the presence of Abeta(1-42). A twofold increase in quenching constant was noted for KI quenching of AEDANS fluorescence emission in the presence of apoE CT domain, indicative of alterations in Abeta conformation upon interaction with apoE CT domain. We propose intermolecular FRET analysis as a discriminating approach to examine apoE/Abeta interaction, a potentially critical factor in early events involved in amyloid formation. PMID- 15880462 TI - Increases in intracellular calcium dephosphorylate histone H3 at serine 10 in human hepatoma cells: potential role of protein phosphatase 2A-protein kinase CbetaII complex. AB - We present evidence that increases in intracellular calcium, induced by treatment with calcium ionophore A23187 or the endoplasmic reticulum calcium-ATPase inhibitor thapsigargin, dephosphorylated histone H3 at serine10 (histone H3 Ser10) in a dose-dependent manner in human hepatoma HepG2 cells. Inhibition of p42/44MAPK, pp90RSK, or p38MAPK did not affect the ability of A23187 to dephosphorylate histone H3-Ser10. This response is significantly blocked by okadaic acid, indicating a requirement for protein phosphatase 2A (PP2A). A23187 increased the activity of PP2A towards phosphorylated histone H3-Ser10. Furthermore, pretreatment with calphostin C, a selective protein kinase C (PKC) inhibitor, blocked A23187-dependent dephosphorylation of histone H3-Ser10, and coimmunoprecipitation analysis showed PP2A association with the PKCbetaII isoform. Unlike untreated cells, coimmunoprecipitated complex from A23187-treated cells showed greater dephosphorylation of histone H3-Ser10 in a PP2A-dependent manner. Inhibition of PP2A increased phosphorylation at Ser660 that determines calcium sensitivity and activity of PKCbetaII isoform, thus supporting a role for intracomplex regulation. Finally, chromatin immunoprecipitation assays following exposure to A23187 and okadaic acid revealed regulatory role of histone H3-Ser10 phosphorylation in selective gene induction. Altogether, our findings suggest a novel role for calcium in modulating histone H3-Ser10 phosphorylation level and led us to propose a model emphasizing PP2A activation, occurring downstream following perturbations in calcium homeostasis, as key event in dephosphorylating histone H3-Ser10 in mammalian cells. PMID- 15880463 TI - Anaerobic metabolism of propionate by polyphosphate-accumulating organisms in enhanced biological phosphorus removal systems. AB - Propionate, a carbon substrate abundant in many prefermenters, has been shown in several previous studies to be a more favorable substrate than acetate for enhanced biological phosphorus removal (EBPR). The anaerobic metabolism of propionate by polyphosphate accumulating organisms (PAOs) is studied in this paper. A metabolic model is proposed to characterize the anaerobic biochemical transformations of propionate uptake by PAOs. The model is demonstrated to predict very well the experimental data from a PAO culture enriched in a laboratory-scale reactor with propionate as the sole carbon source. Quantitative fluorescence in-situ hybridization (FISH) analysis shows that Candidatus Accumulibacter phosphatis, the only identified PAO to date, constitute 63% of the bacterial population in this culture. Unlike the anaerobic metabolism of acetate by PAOs, which induces mainly poly-beta-hydroxybutyrate (PHB) production, the major fractions of poly-beta-hydroxyalkanoate (PHA) produced with propionate as the carbon source are poly-beta-hydroxyvalerate (PHV) and poly-beta-hydroxy-2 methylvalerate (PH2MV). PHA formation correlates very well with a selective (or nonrandom) condensation of acetyl-CoA and propionyl-CoA molecules. The maximum specific propionate uptake rate by PAOs found in this study is 0.18 C-mol/C-mol biomass . h, which is very similar to the maximum specific acetate uptake rate reported in literature. The energy required for transporting 1 carbon-mole of propionate across the PAO cell membrane is also determined to be similar to the transportation of 1 carbon-mole of acetate. Furthermore, the experimental results suggest that PAOs possess a similar preference toward acetate and propionate uptake on a carbon-mole basis. PMID- 15880464 TI - Identification of recurrent chromosomal aberrations in germ cell tumors of neonates and infants using genomewide array-based comparative genomic hybridization. AB - Human germ cell tumors (GCTs) of neonates and infants comprise a heterogeneous group of neoplasms, including teratomas and yolk sac tumors with distinct clinical and epidemiologic features. As yet, little is known about the cytogenetic constitution of these tumors. We applied the recently developed genomewide array-based comparative genomic hybridization (array CGH) technology to 24 GCTs derived from patients under the age of 5 years. In addition, we included seven tumors derived from children and adolescents older than 5 years. In the series from those under the age of 5 years, most teratomas displayed normal profiles, except for some minor recurrent aberrations. In contrast, the yolk sac tumors displayed recurrent losses of 1p35-pter and gains of 3p21-pter and of 20q13. In the GCTs of patients older than 5 years, the main recurrent anomalies included gains of 12p and of whole chromosomes 7 and 8. In addition, gains of the 1q32-qter region and losses of the 6q24-qter and 18q21-qter regions were frequent in GCTs of varied histology, independent of age. We concluded that array CGH is a highly suitable method for identifying recurrent chromosomal anomalies in GCTs of neonates and infants. The recurrent anomalies observed point to chromosomal regions that may harbor novel diagnostic/prognostic identifiers and genes relevant to the development of these neoplasms. PMID- 15880465 TI - Treatment of femoral head necrosis with free vascularized fibula grafting: a preliminary report. AB - Since October 2000, 56 hips in 48 patients with avascular necrosis of the femoral head were treated with free vascularized fibular transplants. The average follow up was about 16 months. The Harris hip scores of all stages were improved during follow-up. Most femoral heads showed improvement (39 hips, 69.6%) or were at least unchanged (14 hips, 25.0%) on X-rays. The results show that a free vascularized fibular graft would be a valuable procedure for femoral head necrosis. By this method, we can avoid or delay progress of the disease, and improve the function of the hip and quality of life. PMID- 15880466 TI - Descending projections of the hamster intergeniculate leaflet: relationship to the sleep/arousal and visuomotor systems. AB - The intergeniculate leaflet (IGL), homolog of the primate pregeniculate nucleus, modulates circadian rhythms. However, its extensive anatomical connections suggest that it may regulate other systems, particularly those for visuomotor function and sleep/arousal. Here, descending IGL-efferent pathways are identified with the anterograde tracer, Phaseolus vulgaris leucoagglutinin, with projections to over 50 brain stem nuclei. Projections of the ventral lateral geniculate are similar, but more limited. Many of the nuclei with IGL afferents contribute to circuitry governing visuomotor function. These include the oculomotor, trochlear, anterior pretectal, Edinger-Westphal, and the terminal nuclei; all layers of the superior colliculus, interstitial nucleus of the medial longitudinal fasciculus, supraoculomotor periaqueductal gray, nucleus of the optic tract, the inferior olive, and raphe interpositus. Other target nuclei are known to be involved in the regulation of sleep, including the lateral dorsal and pedunculopontine tegmentum. The dorsal raphe also receives projections from the IGL and may contribute to both sleep/arousal and visuomotor function. However, the locus coeruleus and medial vestibular nucleus, which contribute to sleep and eye movement regulation and which send projections to the IGL, do not receive reciprocal projections from it. The potential involvement of the IGL with the sleep/arousal system is further buttressed by existing evidence showing IGL efferent projections to the ventrolateral preoptic area, dorsomedial, and medial tuberal hypothalamus. In addition, the great majority of all regions receiving IGL projections also receive input from the orexin/hypocretin system, suggesting that this system contributes not only to the regulation of sleep, but to eye movement control as well. PMID- 15880467 TI - Nuclear condensation of cyclic adenosine monophosphate responsive element-binding protein in discrete murine brain structures. AB - We have directed a polyclonal antibody against an oligo-peptide (123-136) of the transcription factor cyclic AMP responsive element-binding protein (CREB) including the serine residue at 133. Rabbit sera were purified by ammonium sulfate precipitation, followed by affinity chromatography to homogeneity on one dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified antibody not only induced marked supershift of CREB binding, without affecting binding of activator protein-1 on gel retardation electrophoresis, but also differentiated between CREB and CREB phosphorylated at serine133 in brain nuclear fractions on Western blotting. Immunoreactive CREB was detected in both cytosolic and nuclear fractions of discrete murine brain structures but was more highly condensed in cerebellum than in neocortex and hippocampus. Incubation of brain nuclear fractions led to a marked export of immunoreactive CREB in a temperature-dependent manner, whereas the temperature-dependent export activity was significantly lower in cerebellum than in other brain structures. Suppression of general new protein synthesis by cycloheximide (500 mg/kg, i.p.) in vivo resulted in a significant decrease in the nuclear CREB level, with a concomitant increase in the cytosolic level in hippocampus, but not in cerebellum. These results suggest that the nuclear export activity might vary from region to region in murine brains through a hitherto unidentified mechanism other than the nuclear localization signal, to result in different nuclear condensation ratios for subsequent elicitation of differential transcriptional activities by the constitutive transcription factor CREB in the nucleus. PMID- 15880468 TI - Quantitative analysis of the "warm-up" phenomenon in myotonic dystrophy type 1. AB - To quantitate improvement in hand-grip myotonia and muscle strength (i.e., the "warm-up" phenomenon) in myotonic dystrophy type 1 (DM1), six successive, standardized maximum voluntary isometric contractions (MVICs) were recorded on 2 separate days using a computerized isometric hand-grip myometer in 25 genetically confirmed DM1 patients and in 17 normal controls. An automated computer program placed cursors along the declining (relaxation) phase of the MVICs at 90%, 50%, and 5% of peak force (PF) and calculated relaxation times (RTs) between these points. Mean 90% to 5% RT (a measure of myotonia) rapidly declined from 2.5 s in MVIC 1 to 0.8 s in MVIC 6 (warm-up = 1.7 s) in DM1; in controls, it remained 0.4 s for all six MVICs (warm-up = 0). In DM1, 70% of warm-up occurred between MVIC 1 and 2, almost exclusively in the terminal 50% to 5% phase of muscle relaxation. Day 1 warm-up was highly correlated with the severity of myotonia, and with day 2 warm-up. Improvement in myotonia was not accompanied by either transient paresis or improvement in PF. We conclude that, with this testing paradigm: warm-up of myotonia in DM1 can be reliably measured; is proportional to severity of myotonia; occurs rapidly, being most prominent between the first and second grips; mainly results from shortening of the terminal phase of muscle relaxation; and is not accompanied by significant warm-up in force output. PMID- 15880469 TI - 6-pentyl-alpha-pyrone production by Trichoderma harzianum: the influence of energy dissipation rate and its implications on fungal physiology. AB - The influence of the agitation conditions on biomass growth, morphology, carbon metabolism, viability, and 6-pentyl-alpha-pyrone (6PP) production by Trichoderma harzianum were studied in an extractive fermentation system. Batch spore inoculated cultures developed at dissolved oxygen concentrations above 35% of air saturation were carried out in a 14 L bioreactor. The effect of energy dissipation rate over culture performance was assessed using two sets of three Rushton turbines (having different diameters) operated at different agitation speeds. Higher mechanical stress enhanced cellular differentiation (i.e., sporulation), while yielding lower specific growth rates and increased specific CO(2) production rates (CPRs) at relatively constant specific glucose consumption rates. In addition, fungal viability and clump mean diameter decreased gradually at higher energy dissipation rates. 6PP biosynthesis was growth associated and its specific productivity showed a bell-shaped relationship with the energy dissipation rate. T. harzianum physiology was, therefore, strongly influenced by the prevailing hydrodynamic conditions as it triggered cellular metabolism and differentiation shifts. PMID- 15880470 TI - Addition of nitric oxide donor S-nitroso-N-acetylcysteine to selective iNOS inhibitor 1400W further improves contractile function in reperfused skeletal muscle. AB - This study examines the effects of combination therapy with the nitric oxide (NO) donor S-nitroso-N-acetylcysteine (SNAC) and the iNOS inhibitor N-(3 (aminomethyl)benzyl) acetamidine (1400W) on contractile function in reperfused rat skeletal muscle. The right extensor digitorum longus (EDL) muscles of 104 rats were subjected to 3 h of ischemia followed by reperfusion times of 3 h, 24 h, and 7 days. For each time period, rats were further divided into sham operation, control, 1400W only, and 1400W plus SNAC groups. In vitro muscle contractile functional testing was performed in an organ chamber with electrical stimulation. The results showed that twitch and isometric tetanic forces were significantly improved in the 1400W-alone group compared to controls for 24 h and 7 days, but not 3 h of reperfusion. However, all three time periods of reperfusion showed that combination treatment of 1400W + SNAC significantly improved muscle contractile force compared to both control and 1400W-only groups. This corresponded to the decreased tissue necrosis and inflammation seen with combination therapy histologically. Our results demonstrate that combination treatment of 1400W + SNAC promotes functional recovery in reperfused skeletal muscle, supporting that manipulation of NO levels with a NO donor and an iNOS inhibitor is more beneficial than either treatment in isolation. PMID- 15880471 TI - Minor elective surgical procedures using general anesthesia in children with sickle cell anemia without pre-operative blood transfusion. AB - BACKGROUND: Pre-operative red blood cell (RBC) transfusions are often recommended for patients with sickle cell disease (SCD) who require elective surgery under general anesthesia. However, definitive randomized studies demonstrating the benefit of transfusions in this setting have not been conducted. In particular, the merits of transfusion prior to minor or low-risk surgical procedures in children with SCD have not been demonstrated. PROCEDURE: We hypothesized that children with sickle cell anemia (Hb SS) who have minor elective surgical procedures develop few complications even without pre-operative transfusion. We accessed our Comprehensive Sickle Cell Program's Database to identify all such procedures performed during a 13-year period. Medical records were reviewed to characterize the surgical procedure, the use of transfusions, and perioperative complications. RESULTS: Twenty-eight children with Hb SS had a total of 38 minor surgical procedures. No perioperative transfusions were given in 34 of the cases (85%). Five of these 34 surgeries (15%) were associated with minor post-operative complications (fever or transient pain). No post-operative acute chest syndrome was encountered. CONCLUSIONS: Minor or low-risk elective surgical procedures in children with Hb SS may not routinely require pre-operative transfusion. A randomized clinical trial to compare transfusion with no transfusion for minor surgical procedures is needed. PMID- 15880472 TI - Collagen XVII and BPAG1 expression in the retina: evidence for an anchoring complex in the central nervous system. AB - The ectoderm gives rise not only to the skin but also to the entire CNS. This common embryonic lineage suggests that some molecular isoforms might serve analogous functions in both tissues. Indeed, not only are laminins important components of dermal adhesion mechanisms, but they also regulate some aspects of synaptic development in both the CNS and the PNS. In the skin, laminins are part of a hemidesmosome complex essential for basal keratinocyte adhesion that includes collagen XVII (BP180) and BPAG1 (dystonin/BP230). Here, we show that CNS neurons also express collagen XVII and BPAG1 and that these molecules are expressed in the adult and developing retina. In the retina, isoforms of collagen XVII and BPAG1 are colocalized with laminins at photoreceptor synapses and around photoreceptor outer segments; both molecules are expressed by rods, whereas cones express collagen XVII but not BPAG1. Moreover, biochemical data demonstrate that collagen XVII complexes with retinal laminins. We propose that collagen XVII and BPAG1 isoforms may help to anchor elements of the rod photoreceptor cytomatrix to the extracellular matrix. PMID- 15880473 TI - Partial demasculinization of several brain regions in adult male (XY) rats with a dysfunctional androgen receptor gene. AB - The adult rat posterodorsal medial amygdala (MePD) is sexually dimorphic in regional volume and neuronal soma size, both of which are larger in males than in females. This sexual dimorphism is entirely dependent on adult circulating levels of testicular androgens, and both androgen and estrogen treatment can masculinize MePD structure. We examined male rats that are rendered androgen-insensitive by the testicular feminization mutation (tfm) of the androgen receptor (AR) gene to determine how a dysfunctional AR affects this and other brain sexual dimorphisms. In adult wild-type rats, the MePD in males had a greater regional volume, rostrocaudal extent, and soma size than in females. In genetic males, defective ARs affected some but not all of these indices: MePD volume and soma size in tfm males were intermediate between those of wild-type males and females, but the rostrocaudal extent of the MePD was unaffected by the mutation, being as great in tfm males as in wild-type males. Regional volume and soma size in the suprachiasmatic nucleus was reduced in tfm males compared with wild-type males, suggesting that AR normally affects this region in male rats. Interestingly, whereas volume of the sexually dimorphic nucleus of the preoptic area was unaffected by the tfm allele, soma size in this region was reduced in tfm males compared with wild-type males. Although estrogen receptor activation has been shown to be vital for masculinization of the rodent brain, our results indicate that ARs also contribute to this process in several brain regions. PMID- 15880474 TI - Adult female and male zebra finches show distinct patterns of spine deficits in an auditory area and in the song system when reared without exposure to normal adult song. AB - Male songbirds typically require exposure to normal adult conspecific song during development in order to learn a normal song of their own. Females require exposure to conspecific song during development in order to select high-quality, learned song over the incomplete song produced by males reared in isolation. Altering males' opportunity for song learning during development affects the neuroanatomy of brain regions involved in song production (the song system), but in females the neural effects of song learning are unknown. We raised male and female zebra finches (Taeniopygia guttata) with differing amounts of exposure to singing males during development. At 120 days, we Golgi-stained their brains and measured the frequency of dendritic spines in brain areas used in song perception or production. We found that females reared with little or no exposure to song have 31% fewer dendritic spines per unit length of dendrite in caudomedial nidopallium (NCM), a brain area activated by song perception, compared to control females. The deprived females had small deficits in the frequency of spines in HVC, a region activated by song production in males. Males with limited exposure to song had a 24% lower spine density in HVC than controls but only a 10% lower density in NCM. These data support the hypothesis that NCM is important in auditory learning, while HVC is involved in sensorimotor learning, and that these capacities are differentially emphasized in the two sexes. PMID- 15880475 TI - Blood levels of free-PSA but not complex-PSA significantly correlates to prostate release of PSA in semen in young men, while blood levels of complex-PSA, but not free-PSA increase with age. AB - BACKGROUND: The proportion of free- and complex-prostate specific antigen (PSA) in serum is used for differentiating between benign and malignant prostate disease. To further understand the physiological relationship between PSA in seminal plasma and blood, we have analyzed free-PSA (fPSA) and complex-PSA (cPSA) in blood and PSA in seminal plasma in young healthy men. We also compared age related changes of PSA-forms in blood from young versus older men. METHODS: Total PSA (tPSA), fPSA, and cPSA were measured in (i) blood and semen from 289 male conscripts (mean age 18.1 years) and in (ii) blood from a representative population of 1,389 men (mean age 46.5 years) without diagnosis of prostate cancer (PCa) during long-term follow-up. RESULTS: fPSA in serum (r = 0.40, P < 0.0001) but not cPSA (r = 0.09, P = 0.11), correlates to PSA in seminal fluid. fPSA levels in blood in young (geometric mean: 0.20 ng/ml) versus middle-aged men (geometric mean: 0.18 ng/ml) was not different (P = 0.06), whereas cPSA in middle aged men (geometric mean: 0.38 ng/ml) was higher (P < 0.0001) than in young men (geometric mean: 0.28 ng/ml). CONCLUSIONS: fPSA in blood, but not cPSA, is associated to PSA in semen ( approximately 17% co-variation). In blood cPSA, but not fPSA, increase with age in healthy men, which may reflect an increasing incidence of prostate disease. PMID- 15880476 TI - Endogenous and exogenous citrate transport and release in prostatic preparations: semi-polarized two-dimensional cultures of human PNT2-C2 cells and isolated tubules and segments of rat prostate. AB - BACKGROUND: Electrophysiological characterization of normal human prostate epithelial cells showed exogenous trivalent citrate transport (release) to be K(+)-dependent. METHODS: (1) Ussing chamber recordings of short circuit current (SCC) were used to study citrate transport in the same (PNT2-C2) cell line grown on micro-pore filters as a monolayer. (2) Release of endogenous citrate from confluent cultures and tubules and segments of rat prostate was measured using a fluorescence technique. (3) Enzyme-spectrophotometry was employed to detect citrate release from segments of rat prostate. RESULTS: Citrate transport across the PNT2-C2 monolayer was asymmetrical, consistent with release into the lumen side. Fluorescence and/or enzyme-spectrophotometric measurements showed that time dependent citrate release (endogeneous and preabsorbed) occurred from rat prostate (tubules and segments), but not kidney or lung. The release was dependent on extracellular K(+) but not Na(+). CONCLUSIONS: Citrate release from prostatic cells and tissues (rat and human) was K(+)-dependent, consistent with the previous electrophysiological data. PMID- 15880477 TI - Psychological adjustment of parents of pediatric cancer patients revisited: five years later. AB - We investigated the psychological functioning of parents of children suffering from pediatric cancer using a prospective design over a five-year time period. Parents of children diagnosed with cancer participated at diagnosis (T1), six months (T2), twelve months (T3), and five years later (T4, n = 115). Repeated measures ANOVAs were calculated for the three measures of psychological distress (GHQ, SCL-90 and STAI-S) to examine changes over time and gender differences. Independent T-tests were computed to examine differences between the mean scores of the parents at T4 and the norm groups. The effects of health status and earlier levels of distress on T4 functioning were examined using ANOVA and partial correlational analysis. Results showed that levels of reported distress, psychoneurotic symptoms and state anxiety significantly decreased across time to normal levels at T4 except on the GHQ. A significantly higher percentage of parents (27%) than in the norm group (15%) showed clinically elevated scores on the GHQ. Mothers had higher scores than fathers only on state anxiety. Parents of relapsed children reported higher anxiety levels than parents of surviving and deceased children. Psychological functioning at T1 was significantly related to functioning at T4. These results support the conclusion that although parental distress decreases with time, a significant number of parents still suffer from clinical distress after five years. Parents of relapsed children are at risk for long-term psychological problems as are those with higher levels of psychosomatic complaints at diagnosis. PMID- 15880478 TI - Collagen-like triple helix formation of synthetic (Pro-Pro-Gly)10 analogues: (4(S)-hydroxyprolyl-4(R)-hydroxyprolyl-Gly)10, (4(R)-hydroxyprolyl-4(R) hydroxyprolyl-Gly)10 and (4(S)-fluoroprolyl-4(R)-fluoroprolyl-Gly)10. AB - For the rational design of a stable collagen triple helix according to the conventional rule that the pyrrolidine puckerings of Pro, 4-hydroxyproline (Hyp) and 4-fluoroproline (fPro) should be down at the X-position and up at the Y position in the X-Y-Gly repeated sequence for enhancing the triple helix propensities of collagen model peptides, a series of peptides were prepared in which X- and Y-positions were altogether occupied by Hyp(R), Hyp(S), fPro(R) or fPro(S). Contrary to our presumption that inducing the X-Y residues to adopt a down-up conformation would result in an increase in the thermal stability of peptides, the triple helices of (Hyp(S)-Hyp(R)-Gly)(10) and (fPro(S)-fPro(R) Gly)(10) were less stable than those of (Pro-Hyp(R)-Gly)(10) and (Pro-fPro(R) Gly)(10), respectively. As reported by Bachinger's and Zagari's groups, (Hyp(R) Hyp(R)-Gly)(10) which could have an up-up conformation unfavorable for the triple helix, formed a triple helix that has a high thermal stability close to that of (Pro-Hyp(R)-Gly)(10). These results clearly show that the empirical rule based on the conformational preference of pyrrolidine ring at each of X and Y residues should not be regarded as still valid, at least for predicting the stability of collagen models in which both X and Y residues have electronegative groups at the 4-position. PMID- 15880479 TI - Blood flow in the skin of type 1 diabetic patients before and after combined pancreas/kidney transplantation. AB - BACKGROUND: To analyze effects of long-term glucose normalization after pancreas transplantation, different parameters of skin microcirculation were assessed by laser Doppler fluxmetry. METHODS: Forty-two type 1 diabetic patients after successful simultaneous pancreas/kidney transplantation (Group A, median 32.3 months posttransplant), 28 patients with functioning kidney grafts, but insulin therapy (Group B, median 64.9 months posttransplant) and 13 diabetic pretransplant patients (Group C, median 14.2 months on dialysis) were compared with 33 healthy subjects (Group D). Resting blood flow, postocclusive hyperemia, venoarteriolar response on the right foot and decrease in blood flow during cold pressure test on the left finger was assessed. RESULTS: Postocclusive hyperemia, decrease in blood flow during cold pressure test and venoarteriolar response were higher in Group D than in all patient groups. Resting blood flow in Group A was significantly lower than in Groups B and C (following values as median): 3.6 perfusion units (PU) versus 7.4 PU in Group B, p < 0.01 and 12.1 PU in Group C, p < 0.001, respectively, and was not significantly different to controls (Group D, 5.2 PU). Postocclusive hyperemia was higher in Group A than in Groups B and C (266.7% vs 160.0%, p < 0.05 and 79.4% n.s., respectively), but significantly less than in Group D (563.5%). The microangiopathy index-high values reflecting less or no microangiopathy-was significantly higher in Group A than in Groups B and C (11.0 vs 4.3, p < 0.001 and 4.7, p < 0.05, respectively), and was very much comparable to the values in healthy controls (Group D, 10,3). The decrease in blood flow during cold pressure test was higher in Group A compared to Groups B and C (25.2% vs 21.1% and 13.8%, n.s., respectively), but much less than in Group D (65,7%). CONCLUSION: These data suggest an improvement without complete normalization of skin microcirculation by long-term blood glucose normalization achieved by pancreas transplantation. PMID- 15880480 TI - Synthesis and GIAO NMR calculations for some new 4,5-dihydro-1H-1,2,4-triazol-5 one derivatives: comparison of theoretical and experimental 1H and 13C chemical shifts. AB - Four novel 3-alkyl(aryl)-4-(4-methoxycarbonylbenzylidenamino)-4,5-dihydro-1H 1,2,4-triazol-5-ones (2) were synthesized by the reactions of 3-alkyl(aryl)-4 amino-4,5-dihydro-1H-1,2,4-triazol-5-ones (1) with methyl 4-formylbenzoate and characterized by elemental analyses and IR, 1H NMR, 13C NMR and UV spectral data. In addition, isotropic 1H and 13C nuclear magnetic shielding constants of 2 were obtained by the gauge-including-atomic-orbital (GIAO) method at the B3LYP density functional level. The geometry of each compound was optimized using the 6-311G basis set. PMID- 15880481 TI - Removing a bottleneck in the Bacillus subtilis biotin pathway: bioA utilizes lysine rather than S-adenosylmethionine as the amino donor in the KAPA-to-DAPA reaction. AB - In biotin biosynthesis, DAPA aminotransferase encoded by the bioA gene catalyzes the formation of the intermediate 7,8-diaminopelargonic acid (DAPA) from 7-keto-8 aminopelargonic acid (KAPA). DAPA aminotransferases from Escherichia coli, Serratia marcescens, and Bacillus sphaericus use S-adenosylmethionine (SAM) as the amino donor. Our observation that SAM is not an amino donor for B. subtilis DAPA aminotransferase led to a search for an alternative amino donor for this enzyme. Testing of 26 possible amino acids in a cell-free extract assay revealed that only l-lysine was able to dramatically stimulate the in vitro conversion of KAPA to DAPA by the B. subtilis DAPA aminotransferase. The K(m) for lysine and KAPA was estimated to be between 2 and 25 mM, which is significantly higher than the K(m) of purified E. coli BioA for SAM (0.15 mM). This higher requirement for lysine resulted in accumulation of KAPA during fermentation of B. subtilis biotin producing strains. However, this pathway bottleneck could be relieved by either addition of exogenous lysine to the medium or by introduction of lysine deregulated mutations into the production strains. PMID- 15880482 TI - INSPstI polymorphism and prostate cancer in African-American men. AB - BACKGROUND: Both prostate cancer and diabetes mellitus are common diseases in African-American men. High insulin levels and insulin resistance have been implicated in prostate cancer development, which has prompted a recent investigation of a possible role for germline variation in the insulin gene (INS) and prostate cancer risk. METHODS: Four hundred sixty-six African-American men with and without prostate cancer from the Flint Men's Health Study were typed for the INS Pst1 genotype using restriction digest and direct sequencing. An association between the Pst1 genotype and prostate cancer was examined using crude and age-adjusted logistic regression models. RESULTS: African-American men who were homozygous for the INS PstI CC genotype were 1.59 times more likely to be diagnosed with prostate cancer compared to men with the TT or TC genotypes (95% CI = 0.93-2.72). The association appeared stronger among diabetics compared to non-diabetics; however this observation was not statistically significant. CONCLUSIONS: Our study, taken together with the report of Ho et al., suggests that the INS Pst1 CC genotype is associated with prostate cancer risk in African American men. Germline variation in the INS gene should be more fully explored in multiethnic studies to elucidate the molecular variant(s) associated with prostate carcinogenesis. PMID- 15880483 TI - Prolonged survival of rat hindlimb allografts following short-course FK506 and mycophenolate mofetil combination therapy. AB - The immunosuppressive effect of combined therapy using FK506 and mycophenolate mofetil (MMF) was studied in rat limb allotransplantation. Dark Agouti rat donor hindlimbs were orthotopically transplanted into Lewis rat recipients. In total, 38 models of transplantation were performed and divided into 8 groups that were treated individually or in combination with FK506 + MMF therapy. Animals were immunosuppressed for 28 days and then observed for up to 140 days. Graft rejection was evaluated both macroscopically and histologically. Survival times for rat limb allotransplants receiving combination FK506 + MMF therapy were significantly longer than with FK506 or MMF monotherapy, and this was achieved without serious side effects. A histopathological study demonstrated a significantly lower level of rejection with FK506 + MMF combination treatment compared to groups receiving FK506 or MMF monotherapy. Combined FK506 + MMF treatment can prolong the survival of rat limb allografts. PMID- 15880484 TI - X-linked Emery-Dreifuss muscular dystrophy and vacuoles: an immunohistochemical characterization. AB - We report a striking abundance of rimmed vacuoles in two brothers with X-linked Emery-Dreifuss muscular dystrophy (X-EDMD) confirmed by the absence of emerin at the muscular nuclear envelope and by genetic analysis showing a new 2-bp deletion in exon 6 of the STA gene at the Xq28 region. Immunohistochemical analysis of the vacuoles revealed expression of dystrophin but not of merosin in the sarcolemma of rimmed vacuoles and absence of amyloid and membrane attack complex (MAC) deposition either in vacuoles or muscle fibers. The presence of rimmed vacuoles can be a histopathological finding in X-EDMD, and the diagnosis should not be excluded in clinically well-defined EDMD patients because of this finding. PMID- 15880485 TI - Intraneural microstimulation of motor axons in the study of human single motor units. AB - Single motor unit activity has been studied in depth since the first intramuscular electrodes were developed more than 70 years ago. Many techniques have been combined or used in isolation since then. Intraneural motor axon microstimulation allows the detailed study of single motor units in awake human subjects in a manner most analogous to that used in reduced animal preparations. A microelectrode, inserted percutaneously into a peripheral nerve, stimulates the axon of a single alpha-motoneuron at a site remote from the contracting muscle, allowing detailed analyses of the contractile properties of a single motor unit in an otherwise quiescent muscle, that is, without interference of simultaneously active motor units or the presence of an electrode within the muscle. The methods and results obtained using this technique are described and compared to those of other studies of single motor units in human subjects. Differences have been found between human and animal motor units and between motor units of various muscles. Studying human and animal motor units using an analogous technique provides insight into the interpretation of human data when results differ from animal data, and when human motor units cannot be examined in the same way, or at a similar level of detail, as animal motor units. PMID- 15880486 TI - Remote ischemic preconditioning of flaps: a review. AB - Ischemic preconditioning (IP) is defined as a brief period of ischemia ("preclamping") followed by tissue reperfusion, thereby increasing ischemic tolerance for a subsequent longer ischemic period. Several studies showed the effectiveness of classic local IP by preclamping the flap pedicle. There are two temporally and mechanically different types of IP: acute preconditioning, which is induced by preclamping the flap pedicle briefly before flap ischemia, and late preconditioning, induced by a preclamping procedure 24-48 h before flap ischemia. However, both types of local ischemic preconditioning are rarely used clinically, most likely since they can be applied only by invasive means, significantly increase operation time, or even require a second surgical procedure. Several studies from our laboratory showed, in different experimental models, that acute IP, enhancement of flap survival, and improvement of reperfusion microcirculation can be achieved not only by preclamping the flap pedicle, but also by induction of an ischemia/reperfusion event in a body area distant from the flap prior to elevation. This new acute remote IP procedure can be applied without invasive means, using limb tourniquet ischemia briefly before flap ischemia. The effectiveness of acute remote IP was confirmed by other authors in large animal models. Another of our studies showed that late remote IP using a limb tourniquet 24 h before flap ischemia attenuates ischemia/reperfusion in muscle flaps, whereas it was ineffective in adipocutaneous flaps. The exact mechanism of "classic" as well as remote IP is not yet finally determined, although several studies demonstrated that endogenous nitric oxide plays an important role. In summary, the use of a tourniquet to induce limb ischemia before flap ischemia could provide a new, alternative, noninvasive remote IP protocol, although late remote IP might be effective only in muscle flaps. However, the possible future clinical application for late IP is elective flap surgery, whereas acute remote IP could even be used in emergency flaps. PMID- 15880487 TI - Trigemino-solitarii-facial pathway in rats. AB - This study was undertaken to identify premotor neurons in the nucleus tractus solitarii (NTS) serving as relay neurons between the sensory trigeminal complex (STC) and the facial motor nucleus in rats. Trigemino-solitarii connections were first investigated following injections of anterograde and/or retrograde (biotinylated dextran amine, biocytin, or gold-HRP) tracers in STC or NTS. Trigemino-solitarii neurons were abundant in the ventral and dorsal parts of the STC and of moderate density in its intermediate part. They project throughout the entire rostrocaudal extent of the NTS with a strong lateral preponderance. Solitarii-trigeminal neurons were observed mostly in the rostral and rostrolateral NTS. They mainly project to the ventral and dorsal parts of the spinal trigeminal nucleus but not to the principal nucleus. Additional neurons located in the middle NTS were found to project exclusively to the spinal trigeminal nucleus pars caudalis. No solitarii-trigeminal cells were observed in the caudal NTS. In addition, evidence was obtained of NTS retrogradely labeled neurons contacted by anterogradely labeled trigeminal terminals. Second, solitarii-facial projections were analyzed following injections of anterograde and retrograde tracers into the NTS and the facial nucleus, respectively. NTS neurons, except those of the rostrolateral part, reached the dorsal aspect of the facial nucleus. Finally, simultaneous injections of anterograde tracer in the STC and retrograde tracer in the facial nucleus gave retrogradely labeled neurons in the NTS receiving contacts from anterogradely labeled trigeminal boutons. Thus, the present data demonstrate for the first time the existence of a trigemino solitarii-facial pathway. This could account for the involvement of the NTS in the control of orofacial motor behaviors. PMID- 15880488 TI - New criteria for early electrodiagnosis of acute inflammatory demyelinating polyneuropathy. AB - A variety of electrodiagnostic methods are used to confirm the diagnosis of acute inflammatory demyelinating polyneuropathy (AIDP), but difficulties are frequent during the first few weeks of weakness. We compared the nerve conduction studies (NCS) of patients with AIDP to those with critical illness polyneuropathy (CIP), a subacute axonal polyneuropathy. New electrodiagnostic criteria with graded certainty (normal, nondiagnostic, suggestive, highly suggestive, and definite) were designed and applied in a blinded manner to both groups. Among the AIDP patients, 64% met the highly suggestive and definite criteria (specificity 95 100%, P < 0.01), whereas 80% of the CIP group fell in the nondiagnostic criteria (P < 0.001). The relative preservation of the sural sensory response in spite of at least two abnormal sensory NCS in the upper limb suggested acute demyelination (sensitivity 48%, specificity 96%, P < 0.001) and was even more conclusive when associated with absent or prolonged F waves. Motor and sensory response amplitudes were lower in the CIP group, with comparable mean motor and sensory distal latencies and motor conduction velocities. Motor conduction blocks were present in 10% of nerves in AIDP and were not encountered in CIP. The frequency of absent or delayed F waves and absent H reflex was similar in both groups. The correlation coefficient of the cerebrospinal fluid protein concentration with the designed criteria was higher in the AIDP group (r = 0.9). We conclude that a new criterion with graded certainty is of higher specificity in the majority of patients with early AIDP. PMID- 15880489 TI - In vivo interaction of pulmonary intravascular macrophages with activated platelets in microvessels of equine lung after multiple exposures to halothane, isoflurane, and thiamylal: a comparative ultrastructural and cytochemical study. AB - The pulmonary intravascular macrophages (PIMs) of equines contain a unique electron-dense surface coat that is predominantly composed of lipoproteins. A single exposure of inhalatory halothane causes mobilization of the surface coat into the endocytotic system of the PIMs, followed by expansion of the Golgi apparatus and its enrichment with acid phosphatase. Simultaneously, the cells of the lymphocytic series show hyperplasia in the form of mitotic changes inside the microvascular compartment of the lung. Halothane is known to cause acute and chronic hepatotoxicity because of its biotransformation into trifluoroacytelated polypeptides. The present study was designed to examine the comparative effects of reexposures of inhalatory doses of halothane, isoflurane, and the intravenous barbiturate thiamylal sodium in ponies to evoke a stronger response in the PIMs after four exposures at increasing intervals of 1, 2, and 6 weeks. Ultrastructural and cytochemical evidence is presented that halothane induced translocation of the surface coat into the vacuolar system of the PIMs, followed by expansion of the Golgi apparatus and its enrichment with acid phosphatase. The cell membrane was thrown into extraordinary lamellipodial extensions, which enabled the PIMs to interact with platelets within the narrow confines of the pulmonary capillaries. The relationship between PIMs and platelets developed into large platelet aggregates. Isoflurane and thiamylal sodium did not affect the circulating platelets, although the surface coat was translocated into the endolysosomes in both situations. Although isoflurane is a lipid-soluble inhalant anesthetic similar to halothane, it is subject to very little biotransformation after use and in the present model demonstrates no immune response. PMID- 15880490 TI - Limiting iron availability confers neuroprotection from chronic mild carbon monoxide exposure in the developing auditory system of the rat. AB - Iron deficiency and chronic mild carbon monoxide (CO) exposure are nutritional and environmental problems that can be experienced simultaneously. We examined the effects of chronic mild CO exposure and iron availability on auditory development in the rat. We propose that chronic mild CO exposure creates an oxidative stress condition that impairs the spiral ganglion neurons. The CO exposed rat pups had decreased neurofilament proteins and increased copper, zinc superoxide dismutase (SOD1) in the spiral ganglion neurons. We conclude that the increased amount of SOD1 causes an increase in hydrogen peroxide production that allows the Fenton reaction to occur. This reaction uses both iron and hydrogen peroxide to generate hydroxyl radicals and leads to the development of oxidative stress that impairs neuronal integrity. However, rat pups with decreased iron and CO exposure (ARIDCO) exhibited in their cochlea an up-regulation of transferrin, whereas their expression of neurofilament proteins and SOD1 were similar to control. Consequently, reduced iron availability and the normal expression of SOD1 do not promote oxidative stress in the cochlea. By using basal c-Fos expression as a marker for cellular activation we found a significant reduction in c-Fos expression in the central nucleus of the inferior colliculus in iron adequate rat pups exposed to CO. By contrast, rather than being reduced, c-Fos expression in the ARIDCO group is the same as for controls. We conclude that the cochlea of rat pups with normal iron availability is selectively affected by mild CO exposure, causing a chronic oxidative stress, whereas limiting iron availability ameliorates the effect caused by mild CO exposure by averting conditions that facilitate oxidative stress. PMID- 15880491 TI - Alternative model to human skin organ culture: a preliminary study with Leibovitz L15 medium. AB - Organ culture has been used to maintain the three-dimensional structure of the skin and the interaction between melanocytes and keratinocytes, which is essential for melanin production. In the current study we aimed to evaluate the general morphology, viability, and distribution of human melanocytes in a system that uses Leibovitz L15 medium at room temperature. By comparison with human skin explants maintained in Dulbecco's minimum Eagle's medium at 37 degrees C, we found that the skin was better preserved with Leibovitz L15 after 7 days in culture. The addition of 10% fetal bovine serum to this medium did not promote any change. Dividing cells labeled with Ki-67 were visualized at the basal and suprabasal epidermal layers. Retinoic acid was tested at 1 microg/mL and we recorded a reduction of the corneal layer after 48 hours and a complete detachment of the epidermis after 7 days, probably due to a toxic effect in the medium. Melanin and melanocytes were detected by ammoniacal silver and Dopa stainings under light microscopy. We observed that cells were viable throughout the culture period and melanin was distributed in melanocytes and keratinocytes. In conclusion, we suggest that the use of Leibovitz L15 medium at room temperature can be a viable alternative to the normal organ culture of human skin, which is an important system to study the activity and reaction of melanocytes to dermatological products and cosmetics. PMID- 15880493 TI - 4-Aminobiphenyl induces liver DNA adducts in both neonatal and adult mice but induces liver mutations only in neonatal mice. AB - The mechanisms underlying the susceptibility of neonatal mice to genotoxic carcinogens were investigated by analyzing the DNA adducts and mutations induced in the livers of neonatal and adult Big Blue transgenic mice by 4-aminobiphenyl (4-ABP), a potent human and rodent carcinogen. Neonatal and adult mice were treated with a regimen of 4-ABP known to induce tumors in neonatal mice. Animals were sacrificed 1 day after the last treatment for DNA adduct analysis and 8 weeks after the last treatment for analysis of lacI and cII mutant frequency (MF). N-(Deoxyguanosin-8-yl)-4-ABP was the major DNA adduct identified in the livers of the 4-ABP-treated mice and levels of this adduct were significantly higher in treated animals than in the controls for both the neonates and adults. Adduct levels for adult females (44.0 +/- 4.8 adducts/10(6) nucleotides) were higher than in neonatal females (25.9 +/- 2.2 adducts/10(6) nucleotides), while adduct levels in adult males (13.5 +/- 2.0 adducts/10(6) nucleotides) were lower than in neonatal males (33.8 +/- 4.1 adducts/10(6) nucleotides). 4-ABP treatment significantly increased the liver cII MFs in both sexes of neonatal mice but not in adult mice. Sequence analysis of cII mutant DNA revealed that 4-ABP induced a unique spectrum of mutations in neonatal mice, characterized by a high frequency of G:C-->T:A transversion, while the mutation spectrum in 4-ABP-treated adults was similar to that of control mice. Our results indicate that DNA adduct formation by 4-ABP depends as much on sex as it does on age, whereas the conversion of DNA adducts into mutations differed with animal age. These observations suggest that neonates are more sensitive than adults to genotoxic carcinogens because the relatively high levels of cell division in the developing animal facilitate the conversion of DNA damage into mutation. Supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html PMID- 15880494 TI - Prolonged lesional expression of RhoA and RhoB following spinal cord injury. AB - Inhibition of the small GTPase ras homology protein (Rho) or its downstream target, the Rho-associated kinase (ROCK), has been shown to promote axon regeneration and to improve functional recovery following spinal cord injury (SCI) in the adult rat. Here, we have analyzed the expression of RhoA and RhoB following spinal cord injury in order to assess whether Rho is a possible target for late pharmacological intervention. In control spinal cords, RhoA(+) cells were almost absent, whereas RhoB was localized to some ependymal cells, a few microglia, and some dissociated neurons. In injured spinal cords, RhoA(+) and RhoB(+)cells accumulated at perilesional areas and in the developing necrotic core early after injury at day 1. After reaching their maximum levels (RhoA at day 3; RhoB at day 1), RhoA(+) and RhoB(+) cell numbers remained significantly elevated until day 28. In areas remote from the lesion (> or =0.75 mm), a more discrete accumulation of RhoA(+) and RhoB(+) cells was observed, primarily in areas of ongoing Wallerian degeneration. RhoA and RhoB were predominantly expressed by polymorphonuclear granulocytes, ED1(+) microglia/macrophages, oligodendrocytes, some neurons, and swollen axons/neurites. Furthermore, expression was located to lesional, reactive astrocytes and fibroblastoid cells confined to areas of scar formation. Our experiments have determined that most RhoA(+) and RhoB(+) cells (>70%) are of mononuclear origin. The persistent presence of lesional RhoA(+) and RhoB(+) axon/neurite fibers over a period of 4 weeks after injury suggests that Rho inhibition is a putative therapeutic concept also for delayed intervention after SCI. PMID- 15880495 TI - Imaging the switching behavior of superparamagnetic nanoislands by spin-polarized scanning tunneling microscopy. AB - In the past, spin-polarized scanning tunneling microscopy (SP-STM) was mainly applied to static domain configurations that do not vary in time. Here, we show that SP-STM may also be used to image the thermal switching behavior of superparamagnetic nanoislands. Special experimental care has to be taken in order to allow the unambiguous interpretation of the obtained data. Most important, the imaging of superparamagnetic particles requires the use of antiferromagnetic probe tips as the stray field of ferromagnetic tips may modify the sample's intrinsic switching behavior. Our results show that Fe monolayer islands on Mo(110) switch thermally when their area is smaller than 40 nm2. Dipolar coupling between adjacent islands is observed at small inter-particle distance. A pronounced shape dependence is found that confirms existing but yet unverified analytical predictions. The first experiments performed on Fe double-layer islands on W(001) also show thermal switching events, but no clear-cut size dependence is found. PMID- 15880496 TI - Microscopy analysis of bone marrow-derived osteoprogenitor cells cultured on hydrogel 3-D scaffold. AB - Bone marrow contains progenitor cells that are able to differentiate into several mesenchymal lineages, including bone. These cells may also provide a potential therapy for bone repair. The purpose of this study was to select the osteoprogenitor cell subpopulation from bone marrow-derived mesenchymal stem cells (MSCs) and to test the ability of a hydrogel scaffold to support growth and osteogenic differentiation. MSCs isolated from rat femur bone marrow were cultured in DMEM medium supplemented with antibiotics, FCS, and L-glutamine. Osteogenic supplements (dexamethasone, sodium beta-glycerophosphate, and ascorbic acid) were added for one, two or three weeks. A selective subpopulation of osteoprogenitor cells was identified by immunohistochemistry, general morphology, scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS). Committed osteogenic cells were transferred to a 3-D hydrogel scaffold and cultured for an additional week. In standard culture, the osteoprogenitor cells formed cell clusters identified by Alizarin red S staining and by positive osteocalcin immunostaining. The number of osteoprogenitor cells, matrix synthesis, and mineralization increased gradually up to three weeks in culture. Mineral deposition in the matrix analyzed by EDS revealed the presence of calcium and phosphate ions at a Ca/P molar ratio of 1.73 in both the osteogenic cultures and the scaffold osteoprogenitor culture. Histological preparations revealed cell clusters within the hydrogel scaffold and SEM analysis revealed cell clusters attached to the scaffold surface. It is concluded that the hydrogel scaffold can support growth and differentiation of osteogenic cultures including mineralization and can potentially serve as a bone graft substitute containing committed osteoprogenitor cells. PMID- 15880497 TI - Functional and molecular mapping of uncoupling between vascular permeability and loss of vascular maturation in ovarian carcinoma xenografts: the role of stroma cells in tumor angiogenesis. AB - Maintaining homogeneous perfusion in tissues undergoing remodeling and vascular expansion requires tight orchestration of the signals leading to endothelial sprouting and subsequent recruitment of perivascular contractile cells and vascular maturation. This regulation, however, is frequently disrupted in tumors. We previously demonstrated the role of tumor-associated myofibroblasts in vascularization and exit from dormancy of human ovarian carcinoma xenografts in nude mice. The aim of this work was to determine the contribution of stroma- and tumor cell-derived angiogenic growth factors to the heterogeneity of vascular permeability and maturation in MLS human ovarian carcinoma tumors. We show by RT PCR and by in situ hybridization that VEGF was expressed by the tumor cells, while angiopoietin-1 and -2 were expressed only by the infiltrating host stroma cells. Vascular maturation was detected in vivo by vasoreactivity to hypercapnia, measured by BOLD contrast MRI and validated by immunostaining of histologic sections to alpha-smooth muscle actin. Vascular permeability was measured in vivo by dynamic contrast-enhanced MRI using albumin-based contrast material and validated in histologic sections by fluorescent staining of the biotinylated contrast material. MRI as well as histologic correlation maps between vascular maturation and vascular permeability revealed a wide range of vascular phenotypes, in which the distribution of vascular maturation and vasoreactivity did not overlap spatially with reduced permeability. The large heterogeneity in the degree of vascular maturation and permeability is consistent with the differential expression pattern of VEGF and angiopoietins during tumor angiogenesis. PMID- 15880498 TI - Medial vestibular connections with the hypocretin (orexin) system. AB - The mammalian medial vestibular nucleus (MVe) receives input from all vestibular endorgans and provides extensive projections to the central nervous system. Recent studies have demonstrated projections from the MVe to the circadian rhythm system. In addition, there are known projections from the MVe to regions considered to be involved in sleep and arousal. In this study, afferent and efferent subcortical connectivity of the medial vestibular nucleus of the golden hamster (Mesocricetus auratus) was evaluated using cholera toxin subunit-B (retrograde), Phaseolus vulgaris leucoagglutinin (anterograde), and pseudorabies virus (transneuronal retrograde) tract-tracing techniques. The results demonstrate MVe connections with regions mediating visuomotor and postural control, as previously observed in other mammals. The data also identify extensive projections from the MVe to regions mediating arousal and sleep-related functions, most of which receive immunohistochemically identified projections from the lateral hypothalamic hypocretin (orexin) neurons. These include the locus coeruleus, dorsal and pedunculopontine tegmental nuclei, dorsal raphe, and lateral preoptic area. The MVe itself receives a projection from hypocretin cells. CTB tracing demonstrated reciprocal connections between the MVe and most brain areas receiving MVe efferents. Virus tracing confirmed and extended the MVe afferent connections identified with CTB and additionally demonstrated transneuronal connectivity with the suprachiasmatic nucleus and the medial habenular nucleus. These anatomical data indicate that the vestibular system has access to a broad array of neural functions not typically associated with visuomotor, balance, or equilibrium, and that the MVe is likely to receive information from many of the same regions to which it projects. PMID- 15880499 TI - Spin-polarized scanning tunneling microscopy: insight into magnetism from nanostructures to atomic scale spin structures. AB - The system of Fe on W(001) is investigated using spin-integrated as well as spin resolved scanning tunneling microscopy (STM). This study ranges from three dimensional Fe islands down to the Fe monolayer and different growth modes are observed related to the preparation temperature. With scanning tunneling spectroscopy (STS), a layer-dependent electronic structure is observed that can easily be used to assign the local coverage to the investigated sample areas. Spin-resolved measurements of the ferromagnetic layers in the pseudomorphic regime immediately reveal the fourfold magnetic in-plane anisotropy. A direct comparison of the observed arrangement of the domains of the exposed layers shows a rotation of the easy axis from the fourth to the third monolayer and a collinear magnetic alignment of third and second monolayer. This is confirmed by the quantitative analysis of the layer-resolved intensities of differential tunneling conductance. The first monolayer does not show a magnetic component parallel to the surface but has a perpendicular anisotropy. For this layer, measurements with an applied magnetic field prove a c(2x2) antiferromagnetic structure, i.e., a checkerboard arrangement of spins. PMID- 15880500 TI - Atomic scale spin-dependent STM on magnetite using antiferromagnetic STM tips. AB - STM tips made from antiferromagnetic MnNi have been used to investigate the atomic structure of the (001) and (111) surfaces of Fe3O4. The clean (001) surface displays a ( radical2 x radical2)R45 degrees superlattice, which is attributed to charge-ordering on the surface, where Fe(2+)-Fe2+ and Fe(3+)-Fe3+ dimers can be discriminated. The oxygen-terminated (111) surface is characterized by a hexagonal superlattice with a periodicity of 42 A. Oxygen vacancies are observed in atomically resolved images of this superlattice. In the presence of an external magnetic field of 60 mT, a significant change in the atomic corrugation in the topmost oxygen layer around each of these defects is observed. The results on both (001) and (111) surfaces are discussed in terms of possible spin-polarized effects. PMID- 15880501 TI - Suppression of metastasis of human pancreatic cancer to the liver by transportal injection of recombinant adenoviral NK4 in nude mice. AB - NK4, a 4-kringle fragment of hepatocyte growth factor (HGF), is an HGF antagonist that also acts as an angiogenesis inhibitor. NK4 strongly inhibits the infiltration, metastasis, and tumor growth of pancreatic cancer. The aim of our study was to evaluate the antitumor effect of adenovirus-mediated NK4 gene transfer to the liver on hepatic metastasis of pancreatic cancer in vivo. We constructed recombinant adenoviral NK4 (Ad-NK4), which encodes a secreted form of human NK4. Intrasplenic injection of Ad-NK4 induced high and relatively maintained expression of NK4 protein in the liver and suppressed the number and growth of metastatic foci in the liver in a nude mouse model. Microscopically, central necrosis was found even in small metastatic foci in Ad-NK4 treated mice. Immunohistochemical analysis of metastatic tumors showed a remarkable decrease in microvessel density and an increase in the number of apoptotic tumor cells after treatment with Ad-NK4. These results indicate that intraportal injection of Ad NK4 may be a useful therapeutic modality for the clinical control of hepatic metastasis in pancreatic cancer. PMID- 15880502 TI - Central role of gene cooption in neural crest evolution. AB - A bona fide neural crest is a defining feature of vertebrate embryos. Protochordate gene expression patterns indicate that neural crest evolution coincided with the cooption of several transcriptional regulators to the neural plate border of the vertebrate ancestor. Recent cell labeling experiments in ascidians suggest that cells in this domain may have been migratory and thus displayed some neural crest cell-like behavior. Taken together, these data suggest that the recruitment of new genetic pathways conferred novel developmental potentials upon the migratory neural tube cells of the prevertebrate chordate. PMID- 15880503 TI - Pododermal angiogenesis and angioadaptation in the bovine claw. AB - Pododermal microvascularization has been suggested to play a key role in the physiological function of the bovine claw and in the pathogenesis of claw diseases. According to our working hypothesis, angiogenesis plays a central role in the physiological and pathological function of the claw and is induced by the pro-angiogenic vascular endothelial growth factor (VEGF). As a basis for further research, the aim of the present study was to examine the mechanisms of pododermal angiogenesis in the functional adaptation of the microvasculature of the claw in histological serial sections and microcorrosion casts of healthy juvenile and adult claws as well as pathologically altered claws. Scanning electron microscopy of microcorrosion casts allowed assessment of the 3D aspect of pododermal angiogenesis and angioadaptation, and was substantiated by a concomitant examination of a 3D in vitro model of angiogenesis based on cultured bovine microvascular endothelial cells. Particularly in the juvenile, but also in the adult claw, sprouting and intussusceptive angioadaptation was demonstrated and resembled the respective stages of in vitro angiogenesis. Evidence of angiogenic processes was also detected in the pathologically altered claws displaying symptoms of subclinical laminitis and/or the digital dermatitis complex. The detected angioadaptation was visible expression of the increased metabolic demands of the claw caused by the growing body weight load. Angiogenic remodeling of the pododermal angioarchitecture was also the connectional reparative principle in pathologically altered claws. Related research perspectives for prophylaxis and therapy of claw diseases are discussed. PMID- 15880504 TI - Mechanically engraved mica surface using the atomic force microscope tip facilitates return to a specific sample location. AB - By controlling the interaction between the atomic force microscope tip and mica, patterns of different sizes and shape have been produced on the surface of mica. Using these operator-constructed patterns as a reliable marker, the original scanned sample location can be re-located and imaged again on the same mica surface by atomic force microscopy (AFM). This location technique can be used to find the same object again even if the sample was removed from the AFM instrument or the sample was imaged in a different mode. PMID- 15880505 TI - AASLD practice guidelines: Evaluation of the patient for liver transplantation. PMID- 15880506 TI - Perineal noise recording as a non-invasive diagnostic method of urinary bladder outlet obstruction: a study in polyvinyl alcohol and silicone model urethras. AB - AIMS: At present, an invasive pressure flow study is recommended to diagnose urinary bladder outlet obstruction. This method induces the risk of urinary tract infection and urethral trauma. We studied perineal noise recording as an alternative, non-invasive diagnostic method in three flexible/extensible model urethras and two silicone tubes. METHODS: The flexible/extensible model urethras were made of a 10% aqueous solution of polyvinyl alcohol (PVA) and differed in wall- stiffness, the silicone tubes differed in diameter and wall-thickness. Three degrees of obstruction were applied by inflating a cuff placed around the PVA-urethras and by compressing the silicone tubes with an adjustable clamp. Noise, produced during flow, was recorded at three positions distal to the obstruction using a piezoceramic contact microphone. RESULTS: The average amplitude of the noise and the essential frequency of the power spectrum of each noise recording depended significantly on the degree of obstruction, the position of the microphone and the wall-stiffness in PVA-urethras and the diameter in silicone tubes. CONCLUSIONS: Based on the results of this study perineal noise recording shows good potential as an alternative method for diagnosing bladder outlet obstruction. PMID- 15880507 TI - Limb chondrogenesis of the seepage salamander, Desmognathus aeneus (amphibia: plethodontidae). AB - Salamanders are infrequently mentioned in analyses of tetrapod limb formation, as their development varies considerably from that of amniotes. However, urodeles provide an opportunity to study how limb ontogeny varies with major differences in life history. Here we assess limb development in Desmognathus aeneus, a direct developing salamander, and compare it to patterns seen in salamanders with larval stages (e.g., Ambystoma mexicanum). Both modes of development result in a limb that is morphologically indistinct from an amniote limb. Developmental series of A. mexicanum and D. aeneus were investigated using Type II collagen immunochemistry, Alcian Blue staining, and whole-mount TUNEL staining. In A. mexicanum, as each digit bud extends from the limb palette Type II collagen and proteoglycan secretion occur almost simultaneously with mesenchyme condensation. Conversely, collagen and proteoglycan secretion in digits of D. aeneus occur only after the formation of an amniote-like paddle. Within each species, Type II collagen expression patterns resemble those of proteoglycans. In both, distal structures form before more proximal structures. This observation is contrary to the proximodistal developmental pattern of other tetrapods and may be unique to urodeles. In support of previous findings, no cell death was observed during limb development in A. mexicanum. However, apoptotic cells that may play a role in digit ontogeny occur in the limbs of D. aeneus, thereby suggesting that programmed cell death has evolved as a developmental mechanism at least twice in tetrapod limb evolution. PMID- 15880508 TI - Cloning and expression analysis of pos-1 in the nematodes Caenorhabditis briggsae and Caenorhabditis remanei. AB - The Caenorhabditis elegans pos-1 gene encodes a zinc-finger protein that is required for germline specification during embryogenesis. The maternally provided mRNA is translationally regulated both spatially and temporally during early development. We have cloned orthologs of pos-1 from C. briggsae and C. remanei, two Caenorhabditis species that have diverged from C. elegans by approximately 20 40 million years. Two regions in the 3' untranslated region are highly conserved among all three species. We find that the pos-1 RNA is expressed in the hermaphrodite and female gonads of C. briggsae and C. remanei but POS-1 protein is not detected at high levels in C. briggsae until the 2-cell stage of embryogenesis. The protein expression is restricted to the germline precursors of the embryo. We conclude that pos-1 appears to be translationally regulated in C. briggsae as it is in C. elegans and speculate the conserved 3' UTR sequences may be involved. PMID- 15880509 TI - Diagnostic DHPLC Quality Assurance (DDQA): a collaborative approach to the generation of validated and standardized methods for DHPLC-based mutation screening in clinical genetics laboratories. AB - Genetic testing in a clinical diagnostic environment must be subject to rigorous quality control procedures, in order to ensure consistency and accuracy of results. Denaturing high performance liquid chromatography (DHPLC) has become a standard prescreening tool for mutation detection, offering very high efficiency and sensitivity of detection. Despite the relatively simple software-assisted assay setup, DHPLC is a complex assay, and quality control is reliant on ensuring optimal instrument performance, excellent assay design and validation, and sufficient user training and proficiency to interpret results. We describe here a unique collaborative effort by a group of diagnostic clinical genetics laboratories with DHPLC expertise who, together with the manufacturer of one of the most widely used DHPLC platforms, have generated standard operating procedures (SOPs) for instrument operation and maintenance, and for mutation detection by DHPLC. We also describe the validation of a disease-specific SOP for DHPLC assisted mutation screening of the MECP2 gene associated with Rett syndrome. The proposed SOP was validated, and used independently in two laboratories to introduce MECP2 testing. In addition, we provide empirically derived normal ranges for the WAVE System Mutation Standards, which are essential for optimal instrument performance. This effort was initiated to try to standardize DHPLC-based mutation screening procedures across laboratories, and so increase the overall quality of this testing method. This endeavor will thus save each laboratory from having to generate SOPs on their own, which is a lengthy and laborious task. In this respect, we define "generic" SOPs as procedures that are easily adaptable to the individual laboratories' quality systems. PMID- 15880510 TI - Conserved versus innovative features in animal body organization. AB - The origin of evolutionary novelties is a central topic in evolutionary developmental biology (evo-devo) studies. In any new feature, there is a conserved component that is either structural or related to the underlying genetic control, but it is not always obvious what is really new and what is conserved. Nevertheless, disentangling this blending of old and new features is basic to understanding mechanisms of evolutionary change. The origin of arthropod appendages illustrates the complexity in tracing the origin of evolutionary novelties. At the base of the lineage, the main body axis was already segmented and antero-posteriorly patterned, and the genetic tool kit required to form lateral outgrowths was already available. The novelty was possibly the developmental decision to "read" the available axial information and to exploit it for differentiating segmentally patterned and axially segmented appendages. Some important novelties bridge the gap between what have been traditionally distinguished as systemic and local changes. For example, the origin of the body cavities evolved by several animal groups may have been initiated by simple changes in cell-to-cell adhesive properties. Any possible change in an existing ontogenetic pathway has the potential to generate novelties. PMID- 15880511 TI - Topological frustrations in Mn films on Fe(001). AB - The high lateral resolution of spin-polarized scanning tunneling microscopy allows new insights into the spin structure of antiferromagnets on the nanometer range. We demonstrate the capability to image a well-defined in-plane component of the sample spin polarization and discuss the spin structure of antiferromagnetic bct Mn in contact with the ferromagnetic Fe(001) substrate. Mn atoms couple ferromagnetically within a Mn atomic plane, while normal to the surface a layer-wise antiferromagnetic order was found. Magnetic frustrations arise in this system at Fe substrate steps at the interface, where topologically induced 180 degrees domain walls are created in the Mn film. A clear widening of the enforced domain walls with increasing Mn thickness was found. The measured widths could be fitted with a linear function and are explained on the basis of a Heisenberg model. PMID- 15880512 TI - A novel atomic force microscope operating in liquid for in situ investigation of electrochemical preparation of porous alumina. AB - A novel atomic force microscope (AFM) operating in liquid is described in this article. The specially designed AFM probe involves a tip attached to a cantilever, a tip holder, and a circular Plexiglas window. When the probe dives into the fluid, a circular meniscus is established around the Plexiglas window, preventing the tip from being affected or destroyed by surface tension of the liquid. In this setup, the whole scanning probe and the sample can completely dive into fluid. Meanwhile, the probe tip scans over the sample surface when the instrument works. These advantages enable the instrument to scan comparatively large or heavy samples with a high speed. The highest scan rate is about 30 lines/s or 14 s for a 400 x 400-pixel, 3 x 3 microm image. Using the new AFM, we carry out in-situ investigation of the formation processes of porous alumina during electrochemical anodic oxidation. A lead ring and an aluminum foil serve as cathode and anode, respectively. They are entirely immersed in the bath electrolyte, which is oxalic acid solution. During anodic oxidation, the AFM images of the sample surface are successively acquired without elevating the sample out of the solution. Experiments reveal that electrochemical reactions take place soon after the power supply is switched on, and with the progression of anodization, nanostructures of porous alumina gradually occur on the aluminum substrate, finally yielding ordered arrays of nanopores. As a typical example of applications, the results of this work show that the new AFM is an ideal and powerful tool for in-situ observation and study of materials or samples in aqueous solutions. PMID- 15880513 TI - Comparison of genomic damage caused by 5-nitrofurantoin in young and adult mice using the in vivo micronucleus assay. AB - The antibiotic 5-nitrofurantoin (5-NF) has been used widely for the treatment of urosepsis in children during the last 20 years. Recent experimentation suggests the need for reevaluating its genotoxic potential. Because of possible differences in the metabolism and clearance of 5-NF in young and adult animals, we conducted a study to determine whether micronuclei caused by 5-NF were age related. The in vivo micronucleus (MN) assay was performed on 3- and 8-week-old mice given single intraperitoneal injections of 5, 10, and 50 mg/kg 5-NF. Blood samples from the tail vein were taken before injection (baseline) and at 48, 96, 168, and 336 hr (2 weeks) after the treatment. One thousand reticulocytes were analyzed for micronuclei from each animal. Compared to similar baseline values for young and adult mice, 5-NF caused a significant increase in MN frequency in both age groups. The mean MN frequency in the young animals was higher than in the adult animals for each dose and sampling time. MN frequencies remained significantly elevated in young animals even 2 weeks after exposure to 5-NF. The results of the study confirm the genotoxic potential of 5-NF in young and adult animals, and indicate that young animals are more sensitive to the genotoxic effects of 5-NF than adult mice and that the response in young mice persists for a significantly longer time. These findings may be related to poorly developed mechanisms of xenobiotic detoxification and renal elimination in young animals. PMID- 15880514 TI - Interaction of statisticians with DMCs. PMID- 15880515 TI - Medication refill logistics and refill adherence in HIV. AB - PURPOSE: Strict adherence to antiretroviral therapy is instrumental in viral suppression and treatment success. The relation between pharmacy-based factors and treatment adherence has been under-explored. We aimed to determine whether different medication refill mechanisms were associated with differences in antiretroviral refill adherence. METHODS: We conducted a retrospective cohort study of 110 HIV-infected subjects on standard antiretroviral regimens for >or=3 months cared for at the Philadelphia Veterans' Affairs Medical Center HIV clinic. The primary outcome was a pharmacy-based measure of antiretroviral refill adherence over the 3 months of treatment immediately prior to the study date. RESULTS: The group obtaining refills at the pharmacy had lower adherence [80% (interquartile range (IQR), 69-99%)] than the group obtaining refills via pill organizers dispensed by a pharmacist [99% (IQR, 97-100%), p=0.003] and the group obtaining refills via mail order [91% (IQR, 79-100%); p=0.04]. CONCLUSIONS: Mail ordering and pharmacists dispensing refills in pill organizers may each be effective strategies for improving medication adherence, although they target different barriers and differ in their degree of intensity. Each should be considered for adherence interventions in HIV and further studied in other disease and treatment settings. PMID- 15880516 TI - Recent advances in atomic-scale spin-polarized scanning tunneling microscopy. AB - The Mn3N2 (010) surface has been studied using spin-polarized scanning tunneling microscopy at the atomic scale. The principle objective of this work is to elucidate the properties and potential of this technique to measure atomic-scale magnetic structures. The experimental approach involves the use of a combined molecular beam epitaxy/scanning tunneling microscopy system that allows the study of atomically clean magnetic surfaces. Several key findings have been obtained. First, both magnetic and non-magnetic atomic-scale information has been obtained in a single spin-polarized image. Magnetic modulation of the height profile having an antiferromagnetic super-period of c = 12.14 A (6 atomic rows) together with a non-magnetic superstructure having a period of c/2 = 6.07 A (3 atomic rows) was observed. Methods of separation of magnetic and non-magnetic profiles are presented. Second, bias voltage-dependent spin-polarized images show a reversal of the magnetic modulation at a particular voltage. This reversal is clearly due to a change in the sign of the magnetic term in the tunnel current. Since this term depends on both the tip's as well as the sample's magnetic local density of states, the reversal can be caused by either the sample or the tip. Third, the shape of the line profile was found to vary with the bias voltage, which is related to the energy-dependent spin contribution from the 2 chemically inequivalent Mn sites on the surface. Overall, the results shown here expand the application of the method of spin-polarized scanning tunneling microscopy to measure atomic-scale magnetic structures. PMID- 15880517 TI - Evaluation of sample spin-polarization from spin-polarized scanning tunneling spectroscopy experiments. AB - Spin-polarized scanning tunneling microscopy has produced a great amount of images presenting magnetic contrast between different magnetic domains with an unsurpassed spatial resolution but getting values like the surface polarization has proven to be a difficult task. We will discuss in detail how to extract this information for the case of manganese layers grown on Fe(001) whiskers. Mn layers adopt a body-centered-tetragonal (bct) structure when they are grown on the Fe(001) surface at room temperature. The Mn layers show an antiferromagnetic coupling between the layers. Comparing our spin-polarized scanning tunneling spectra measured with Fe-coated W tips with spin-resolved band structure calculations, we are able to find the value of the sample surface polarization. Also discussed is a method to change the tip magnetization. Finally, the magnetic structure around a screw dislocation on the surface is reviewed. PMID- 15880519 TI - Intrarectal ground electrode improves the reliability of motor evoked potentials recorded in the anal sphincter. AB - Motor evoked potentials (MEPs) can be recorded in the external anal sphincter in response to magnetic stimulation of the cerebral cortex or sacral roots. However, the magnitude of the stimulus artifact may alter the reliability of anal MEP recording. An intrarectal ground electrode substantially reduces stimulus artifact and technical failure, improving MEP latency determination to sacral root stimulation in particular. PMID- 15880520 TI - Diversity matters: dynamic simulation of distributed bacterial states in suspended growth biological wastewater treatment systems. AB - A MATLAB-based program was developed to simulate the distributions of states and behaviors of microbial storage product-accumulating bacteria in suspended growth systems. Currently available computer simulators of these systems predict dynamic behaviors by numerically solving differential biokinetic equations using average, or "lumped" system states (e.g., average microbial storage products concentrations). However, individual bacterial states are expected to diverge from average values, in part because individuals can have different hydrodynamic histories in terms of their residence times in upstream completely mixed flow reactors. The distributed state simulation program presented in this paper (DisSimulator 1.0) tracks individual bacteria as they move through a completely mixed reactor system. The program was evaluated for competition between polyphosphate-accumulating organisms (PAOs) and non-polyphosphate-accumulating heterotrophs in an enhanced biological phosphorus removal (EBPR) system for treatment of domestic wastewater. For identical systems and process conditions, simulations accounting for distributed states predicted larger anaerobic and aerobic solids residence time requirements for successful EBPR than did simulations using the lumped approach. One reason for this was that distributed simulations predicted large numbers of the PAOs were essentially inactive due to depleted or maximized storage product contents, while the lumped simulations predicted homogenous, 100% active PAO populations. Characteristic state profile shapes developed rapidly and were stable as total population numbers changed. Lumped state assumptions were demonstrated to produce large errors in predictions of EBPR system performance, and so consideration of distributed states may improve the accuracy of microbial storage products-based process simulations in systems with completely mixed hydrodynamics. PMID- 15880521 TI - Introduction: fifteen years of spin-polarized scanning tunneling microscopy. PMID- 15880522 TI - Pontine-wave generator activation-dependent memory processing of avoidance learning involves the dorsal hippocampus in the rat. AB - The aim of this study was to test the hypothesis that the dorsal hippocampus plays a critical role in pontine-wave (P-wave) generator activation-dependent memory processing of two-way active avoidance (TWAA) learning. To achieve this objective, rats were given small bilateral lesions in the CA1, dentate gyrus (DG), or CA3 region of the dorsal hippocampus by microinjecting ibotenic acid. After recovery, lesioned and sham-lesioned rats were trained on a TWAA learning paradigm, allowed a 6-hr period of undisturbed sleep, and then were tested on the same TWAA paradigm. It was found that lesions in the CA3 region impaired retention of avoidance learning. Conversely, lesions in the CA1 and DG regions had no effect on TWAA learning retention. None of the groups showed any changes in the baseline sleep-wake cycle or in the acquisition of TWAA learning. All rats showed increased rapid eye movement (REM) sleep and increased REM sleep P-wave density during the subsequent 6-hr recording period. Impaired retention in the CA3 group occurred despite an increase in REM sleep and P-wave density, suggesting that during REM sleep, the P-wave generator interacts with the CA3 region of the dorsal hippocampus to aid in consolidation of TWAA learning. The results of the present study thus demonstrate that P-wave generator activation dependent consolidation of memory requires an intact CA3 subfield of the dorsal hippocampus. The results also provide evidence that under mnemonic pressure, the dorsal hippocampus may not be involved directly in regulating the sleep-wake cycle. PMID- 15880523 TI - Evidence that one subset of anaplastic thyroid carcinomas are derived from papillary carcinomas due to BRAF and p53 mutations. AB - BACKGROUND: Anaplastic thyroid carcinoma (ATC) is the most lethal form of thyroid neoplasia and represents the end stage of thyroid tumor progression. In the current study, genetic alterations in a panel of ATC were profiled to determine the origins of ATC. METHODS: Eight ATC were analyzed for BRAF mutation at codon 599 by using mutant-allele-specific polymerase chain reaction (PCR) and DNA sequencing of the PCR-amplified exon 15. RAS mutation (HRAS, KRAS, and NRAS) at codons 12, 13, and 61 was analyzed by direct sequencing of PCR-amplified exons 1 and 2 of the RAS gene. RET/PTC rearrangements and p53 mutation were monitored by immunohistochemical (IHC) staining by anti-RET antibodies and an anti-p53 mAb, respectively. RESULTS: BRAF was mutated in 5 of the 8 ATCs tested. Histologic examination revealed that 4 of these 5 BRAF-mutated ATCs contained a PTC component, suggesting that they may be derived from BRAF-mutated PTC. Of the 3 ATCs with wild-type BRAF, 2 had spindle cell features; one had follicular neoplastic characteristics mixed with papillary structures. Analysis of RAS mutation revealed only an HRAS mutation at codon 11, due to the transversion of GCC to TCC in one ATC with wild-type BRAF. This leads to the substitution of valine to serine. IHC analysis of RET/PTC rearrangements revealed no positive staining of RET in any of 8 ATCs, suggesting that these ATCs are not derived from RET/PTC- rearranged PTC. In contrast, IHC analysis of p53 mutation revealed that p53 was detected in the nuclei of 5 of 5 BRAF-mutated ATCs and 2 of 3 ATCs with wild-type BRAF. p53 staining was present only in anaplastic thyroid tumor cells but not in neighboring papillary thyroid tumor cells. CONCLUSIONS: These results suggest that many ATCs with papillary components are derived from BRAF-mutated PTC, because of the addition of p53 mutation. PMID- 15880524 TI - A variant of the alpha-methyl-acyl-CoA racemase gene created by a deletion in exon 5 and its expression in prostate cancer. AB - BACKGROUND: Alpha-methylacyl-CoA racemase (AMACR) is a mitochondrial and peroxisomal enzyme that is overexpressed in prostate cancer. Alternatively spliced variants of AMACR have recently been reported, however, their role in prostate cancer pathogenesis is unclear. METHODS: Using PCR techniques we have identified a novel variant of AMACR. RESULTS: This transcript arises by an alternative splicing event in the 5th exon of the gene whereby a 749 base sequence is deleted causing a shift in the reading frame. The protein encoded by this transcript has a predicted molecular weight of 43,833 kDa and a pI of 7.01 and therefore differs in size and physical characteristics from the main form of AMACR. The carboxyl terminus of this variant does not contain the peroxisomal targeting signal found in the main form of AMACR. Using real time PCR it was demonstrated that this transcript also occurs in normal prostate tissue and is elevated in prostate cancer. Coordinate expression of this transcript with the other forms of AMACR was shown. This transcript was expressed as a FLAG fusion protein in Cos-7 cells and probed with relevant antibodies. CONCLUSION: A deletion event in exon 5 of the AMACR gene creates a novel transcript that is coordinately expressed with the other forms of AMACR but with different biochemical characteristics. (c) 2005 Wiley-Liss, Inc. PMID- 15880525 TI - Expression of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 in human prostate cancer is associated with regional lymph node metastasis. AB - BACKGROUND: Vascular endothelial growth factor C (VEGF-C) and its receptor, VEGFR 3, have been implicated as important factors in the formation of lymphatic vessels and recent evidence suggests that tumor lymphangiogenesis promotes lymphatic metastasis. METHODS: The expression of VEGF-C and VEGFR-3 was examined in 22 human prostate cancer specimens with immunohistochemistry. A semi quantitative scoring system was used for evaluation of staining. RESULTS: Expression of VEGF-C was stronger in prostate cancer areas in comparison to adjacent benign glands. In addition, patients with lymph node metastases had a significantly higher expression of VEGF-C than patients without lymph node metastases. Interestingly, VEGFR-3 was expressed in malignant prostate epithelial cells and its expression was significantly higher in the lymph node positive group compared to the lymph node negative group. CONCLUSIONS: The results of the present study indicate that increased expression of VEGF-C and VEGFR-3 play a role in prostate cancer progression and in metastasis to regional lymph nodes. PMID- 15880526 TI - Selection of knee radiographs for trials of structure-modifying drugs in patients with knee osteoarthritis: a prospective, longitudinal study of Lyon Schuss knee radiographs with the definition of adequate alignment of the medial tibial plateau. AB - OBJECTIVE: The quality of medial tibial plateau (MTP) alignment, which is assessed by measuring the distance between the anterior and posterior margins (intermargin distance [IMD]) of the tibial plateau, and the reproducibility of alignment in serial radiographs are suggested to be key elements in determining the accuracy and sensitivity to change in knee radiographs in patients with tibiofemoral osteoarthritis (OA). We evaluated the influence of both MTP alignment and radiograph superimposition on the sensitivity to change in radiographic joint space narrowing (JSN) in knee OA. METHODS: The study group comprised 106 patients with knee pain (73 with OA). Lyon schuss radiographic images of the knee were obtained twice (at baseline [month 0] and 12 months later), using a standardized radiographic procedure. Computerized measurement of the IMD for the assessment of MTP alignment was compared with the grading of MTP alignment by 2 observers using a 5-point scale (excellent, good, fair, poor, bad). To obtain the rate of JSN, computerized measurement of the joint space width was performed at month 0 and month 12. The sensitivity of the joint space width to change over 1 year was evaluated by the standardized response mean (SRM). RESULTS: The mean (+/-SD) IMD was 1.2 +/- 0.9 mm. The correlation between scoring and computer measurement of MTP alignment was highly significant. The cutoff value for satisfactory alignment (excellent or good) was an IMD of 1.2 mm at month 0 and/or month 12. CONCLUSION: The quality of MTP alignment at both baseline and the end point highly influences the sensitivity to change in radiographic JSN in knee OA. To obtain relevant data, only radiographs showing an IMD of /=4 amino acid residues from the N-terminus failed to induce chemotaxis and calcium influx by CX3CR1-expressing cells. Of these, the most potent antagonist (Fkn-AT) lacked the 4 N-terminal amino acid residues. Fkn expression in the glomerulus was significantly increased in 12-week-old MRL/lpr mice. Expression was localized predominantly in the glomerular endothelial cells, but was occasionally observed in the mesangial cells and, to a lesser extent, in the interstitial microvasculature. Inoculation of MRL/lpr mice with Fkn-AT before the onset or during the early stages of lupus nephritis significantly reduced glomerular hypercellularity, glomerulosclerosis, crescent formation, and vasculitis compared with control mice. This seemed to be due to a marked reduction in macrophage accumulation. In contrast, Fkn antagonist did not affect pneumonitis, sialadenitis, lymphadenopathy, or splenomegaly. CONCLUSION: We prepared a novel potent Fkn antagonist and demonstrated its ability to delay the initiation and ameliorate the progression of lupus nephritis. This agent may therefore provide a new therapeutic approach to lupus nephritis. PMID- 15880600 TI - Analysis of immune reconstitution after autologous bone marrow transplantation in systemic sclerosis. AB - OBJECTIVE: To analyze hematopoietic and immune reconstitution after autologous hematopoietic stem cell transplantation (HSCT) in 7 patients with systemic sclerosis (SSc). METHODS: Two groups of patients were retrospectively constituted according to whether they had a favorable clinical response (group A; n = 4) or no response or a relapse of disease (group B; n = 3) after HSCT. Immune reconstitution was analyzed every 3 months using lymphocyte immunophenotyping, alpha/beta T cell receptor (TCR) diversity analysis, and ex vivo thymic function analysis by quantification of TCR rearrangement excision circles (TRECs). RESULTS: Patients had similar characteristics at study entry, except for a lower modified Rodnan skin thickness score (P = 0.03) and a lower Health Assessment Questionnaire score (P = 0.05) in group A than in group B. The number of reinjected cells and the time to hematopoietic reconstitution were similar in both groups. The absolute numbers of CD19+ and CD20+ B cells were lower in group A than in normal controls (P < 0.05) and within the normal range in group B. Absolute numbers of T and natural killer lymphocytes were normal before HSCT. Numbers of CD3+ cells remained low thereafter. Numbers of CD8+ cells were back to normal 3 months after HSCT in both groups. B cell counts were low until 6 months after HSCT in group A and stayed in the normal range in group B. The CD3+ defect was sustained in group A, with an opposite trend and a faster CD4+ reconstitution profile in group B. The T cell repertoire was skewed before and until 1 year after HSCT, with shared expansions before and after transplant in a given individual. TREC values correlated negatively with C-reactive protein levels (r(s) = -0.41, P = 0.001) and positively with CD19+ (r(s) = 0.35, P = 0.001) and CD20+ (r(s) = 0.34, P = 0.002) lymphocyte counts. CONCLUSION: B and T lymphocyte populations remained disturbed for at least 1 year after HSCT in SSc patients, which may reflect the persistence of an underlying disease mechanism. PMID- 15880601 TI - Additive bone-protective effects of anabolic treatment when used in conjunction with RANKL and tumor necrosis factor inhibition in two rat arthritis models. AB - OBJECTIVE: To investigate whether the bone-preserving effects of a RANKL antagonist or a tumor necrosis factor (TNF) antagonist could be further improved by the addition of a bone anabolic agent in inflammatory arthritis. METHODS: Lewis rats with either adjuvant-induced arthritis (AIA) or collagen-induced arthritis (CIA) were treated for 10 days with PEGylated soluble tumor necrosis factor receptor type I (PEG sTNFRI), interleukin-1 receptor antagonist (IL-1Ra), osteoprotegerin (OPG), parathyroid hormone (PTH), or combinations of these agents starting on day 4 after disease onset. Treatment effects were assessed clinically, radiologically, and histologically, and by morphometry for the extent of paw swelling, bone erosive changes, and synovial inflammation. RESULTS: Paw swelling and synovial inflammation were significantly inhibited by PEG sTNFRI in AIA and CIA, and by IL-1Ra in CIA. OPG and PTH had no significant effect on these parameters. Analysis of bone erosion revealed a significant bone-sparing effect of monotherapy with PEG sTNFRI or OPG in both models, whereas IL-1Ra was only effective in CIA. PTH treatment alone did not show a bone-protective effect in either model. With the combination of PEG sTNFRI and PTH, erosion scores (-74% in AIA and -61% in CIA versus controls) were significantly lower than those elicited by PEG sTNFRI alone (-41% and -29%, respectively, versus controls). Similar results were also obtained with the combination of OPG and PTH (-88% in AIA and 73% in CIA, compared with -70% and -55%, respectively, with OPG monotherapy). Coadministration of IL-1Ra and PTH had no synergistic bone-sparing effect. Morphometric analysis revealed that the addition of PTH to PEG sTNFRI or OPG resulted in higher bone volume and higher osteoblast numbers in both AIA and CIA. CONCLUSION: The bone-protective effects resulting from RANKL or TNF antagonism can be further improved by the addition of a bone anabolic agent. PMID- 15880602 TI - Association between single-nucleotide polymorphisms in the SEC8L1 gene, which encodes a subunit of the exocyst complex, and rheumatoid arthritis in a Japanese population. AB - OBJECTIVE: To identify rheumatoid arthritis (RA) susceptibility genes in a Japanese population by conducting a large-scale case-control association analysis and linkage disequilibrium (LD) mapping on chromosome 7q31-34, a candidate susceptibility locus identified in a preliminary genome-wide scan in 53 Japanese families, using single-nucleotide polymorphisms (SNPs). METHODS: We prepared 728 dense, evenly spaced SNPs with a minor allele frequency >0.15 in each gene locus on chromosome 7q31-34. Using these SNPs, a 2-stage case-control analysis was performed on 760 RA patients (157 men and 603 women) and 806 non-RA controls (189 men and 617 women). Haplotypes and LD mapping results were assessed based on SNP genotypes in 380 controls. RESULTS: Forty-eight SNPs showed allele associations (P < 0.05) in the first set of DNA samples (380 RA cases and 380 non-RA controls; first-stage analysis). For 4 of the SNPs in the SEC8L1 gene, the association was replicated (P < 0.05) in the second, independent set of DNA samples (an additional 380 RA cases and 380 non-RA controls; second-stage analysis). When data from the 2 groups were combined, the most significant allele association was observed with SNP 441, an intronic SNP of the SEC8L1 gene (P = 0.000059). The SEC8L1 SNPs with significant allele associations were all located in a single conserved LD block (block 4). Haplotype analysis revealed the disease-risk (P = 0.0015) and disease-protective (P = 0.0000062) haplotypes. Resequencing of coding exons within block 4 did not identify any nonsynonymous SNPs. Real-time quantitative polymerase chain reaction revealed that SEC8L1 was expressed ubiquitously in human tissues, including fibroblast-like synoviocytes from RA patients. CONCLUSION: Our locus-wide association and LD analyses identified intronic SNPs and haplotypes in the SEC8L1 gene that are strongly associated with RA. We propose that SEC8L1, which encodes a component of the exocyst complex, is a candidate susceptibility gene for RA in the Japanese population. PMID- 15880603 TI - Circular dichroism spectroscopic study of non-covalent interactions of poly-L glutamic acid with a porphyrin derivative in aqueous solutions. AB - The interactions of poly-L-glutamic acid and a cationic porphyrin derivative in aqueous solutions were studied by the combination of vibrational circular dichroism (VCD) and electronic circular dichroism (ECD) spectroscopies. It was found that non-covalent interactions between both agents influence the structure of the polymeric matrix and the guest porphyrins and vice versa, but the physico chemical properties of the solutions, especially the pH and the relative permittivity of the solvent, play a key role in the structure of the polypeptide part of the formed complexes. It was shown that the interaction with porphyrins prevents the precipitation of poly-L-glutamic acid in aqueous solution at acidic pH. In special conditions, the porphyrins attached to the polypeptide probably possess face-to-face interaction as demonstrated by the enhancement of the characteristic ECD signal and the appearance of sidebands on its short and long wavelength sides. PMID- 15880604 TI - Statistical design of experiments as a tool in mass spectrometry. AB - This Tutorial is an introduction to statistical design of experiments (DOE) with focus on demonstration of how DOE can be useful to the mass spectrometrist. In contrast with the commonly used one factor at a time approach, DOE methods address the issue of interaction of variables and are generally more efficient. The complex problem of optimizing data-dependent acquisition parameters in a bottom-up proteomics LC-MS/MS analysis is used as an example of the power of the technique. Using DOE, a new data-dependent method was developed that improved the quantity of confidently identified peptides from rat serum. PMID- 15880605 TI - A comprehensive study of optimal conditions for naked plasmid DNA transfer into skeletal muscle by electroporation. AB - Efficient gene transfer is a key factor in gene therapy. Reducing the damage caused by gene transfer to muscle by electroporation is very important for its clinical application. Extensive investigation of optimal conditions for gene transfer by electroporation is required. The parameters used for electroporation, including plasmid concentration; injection volume; the plasmid dose of the injection; the concentration of saline media; the size of plasmid DNA; the age of the mice; the lag time between plasmid injection and electroporation; and the effect of repeated gene transfer by electroporation, were systematically investigated in the present study. The efficiencies of gene transfer by electroporation in normal and rodent models of diabetes were also evaluated. We found that electroporation used for non-viral gene transfer could be repeated in the same place in the muscle, but the expression efficiency was closely related to the muscle damage. Increasing pulse times could enhance the efficiency of gene transfer with a lower strength of electric field. It was better to use a higher plasmid concentration than to use a larger dose of plasmid and repeated injection to achieve a high level of transgene expression. Optimal conditions varied in different animal models, being milder for diabetic mice than for normal mice, and it was also shown that the conditions that worked well on these small rodents were not necessarily suitable for larger animals. Our results provide a comprehensive view of the factors that affect the efficiency of gene transfer into skeletal muscle by electroporation. PMID- 15880606 TI - Assessment of optimal transduction of primary human skin keratinocytes by viral vectors. AB - BACKGROUND: Genetically modified keratinocytes generate transplantable self renewing epithelia suitable for delivery of therapeutic polypeptides. However, the variety of viral vectors and experimental conditions currently used make fragmented or contradictory the information on the transduction efficiency of the human primary keratinocytes. To compare the suitability of the most currently used viral vectors for efficient gene transfer to human keratinocytes, we have performed a comparative study using a panel of recombinant constructs. METHODS: For each vector, the transduction efficiency and the persistence of the transgene expression were quantified by fluorescence microscopy and flow cytometry analysis of the infected cells. RESULTS: We show that: (1) canine and human adenoviral vectors achieve a highly efficient but transient transduction of both primary and immortalized keratinocytes; (2) the adenovirus-associated virus (AAV) vectors transduce immortalized keratinocytes, albeit with a short-lived gene expression (<4 days), but fail to infect primary keratinocytes; and (3) under appropriate conditions, the oncoretroviral and lentiviral vectors can permanently transduce up to 100% of primary keratinocytes, but the highly clonogenic keratinocytes are more efficiently targeted by lentiviral vectors. CONCLUSIONS: Therefore, AAV vectors are unsuitable to transduce primary keratinocytes, while human and canine adenoviral vectors appears to be appropriate to achieve short-term delivery of therapeutic products. Recombinant retroviruses provide sustained expression of the transgene, but the lentiviral vectors are the most suitable for ex vivo gene therapy because of their ability to transduce clonogenic primary keratinocytes. PMID- 15880607 TI - Regionalization of axial skeleton in the lungfish Neoceratodus forsteri (Dipnoi). AB - Differentiation of the axial skeleton into distinct regions, once thought to be characteristic of the Tetrapoda, also occurs in the actinopterygian Danio rerio. In these taxa, the boundary between the cervical-thoracic regions correlates with Hoxc6 expression and morphological features such as position of the pectoral fin and associated nerves, and the absence of ribs. In the lungfish Neoceratodus, a member of the extant sister taxon to the Tetrapoda, the first vertebral element to chondrify is situated well posterior to the skull, developing from somites 6 and 7 (6/7) and associated with an enlarged cranial rib and nerves innervating the pectoral fin. Two vertebral elements develop later and more anteriorly, associated with somites 4/5 and 5/6. These three elements become incorporated into the occipital region of the skull during Neoceratodus ontogeny, until the cranial rib itself articulates to the rear of the skull. These features of early development indicate a regionalization of the Neoceratodus vertebral column: the cranial rib marks the boundary between the cervical and thoracic regions, the two more anterior vertebrae lacking ribs represent the cervical region, while somites 1-4 (cranial half), lacking any vertebral development, represent the occipital region. However, the cervical region of the vertebral column is effectively lost during ontogeny of Neoceratodus. A recognizable cervical region in the tetrapod vertebral column, as in zebrafish, suggests that cervical vertebrae are not incorporated into the skull but maintained as distinct elements of the column, representing an important shift in relative developmental timing and the influence of heterochrony in this region during the fish-tetrapod transition. PMID- 15880608 TI - Long-term follow-up of peginterferon and lamivudine combination treatment in HBeAg-positive chronic hepatitis B. AB - We have previously demonstrated that combination peginterferon and lamivudine treatment has superior antiviral efficacy to lamivudine monotherapy in chronic hepatitis B. In this study, we investigated the long-term posttreatment virological response to this combination treatment. Sustained virological response of patients who completed 32-week peginterferon and 52-week lamivudine combination treatment was compared to patients who completed 52-week lamivudine monotherapy. Sustained response was defined as sustained hepatitis B e antigen (HBeAg) loss and HBV DNA < 100,000 copies/mL from treatment cessation until the end of follow-up. Forty-eight patients receiving combination treatment and 47 patients receiving lamivudine monotherapy were studied. The posttreatment follow up of patients who received combination treatment was 117 +/- 34 weeks and that of patients receiving lamivudine monotherapy was 124 +/- 29 weeks. At the end of treatment, HBeAg loss occurred in 63% of patients in the combination group and 28% of patients in the lamivudine group (P = .001). The probabilities of sustained response for combination treatment and lamivudine monotherapy were 33% and 13% at week 24, 31% and 11% at week 52, and 29% and 9% at week 76, respectively (log-rank test, P = .0015). No patients developed virological relapse after week 76 until the last visit in either treatment group. All sustained responders had no biochemical relapse (alanine aminotransferase [ALT] > 2 times upper limit of normal) during follow-up. Among the non-sustained responders, biochemical relapse occurred in 32 patients (94%) in the combination group and 38 patients (88%) in the lamivudine group, respectively. In conclusion, combination treatment of peginterferon and lamivudine has a higher sustained virological response than lamivudine monotherapy up to 3 years after treatment. PMID- 15880609 TI - EGFR and Her-2 regulate the constitutive activation of NF-kappaB in PC-3 prostate cancer cells. AB - BACKGROUND: The mechanism through which NF-kappaB (NF-kappaB) is constitutively activated in prostate cancer cells remains unclear. We investigated whether members of the ErbB family of epidermal growth factor receptors (EGFR) are involved in the constitutive activation of NF-kappaB in prostate cancer cell lines. METHODS AND RESULTS: EGFR, Her-2, and ErbB3 are expressed and constitutively activated in PC-3, DU145, and LNCaP prostate cancer cells lines. Using several pharmacological ErbB inhibitors, we demonstrate that EGFR and Her-2 are involved in the constitutive activation of NF-kappaB in PC-3 cells through two different mechanisms. EGFR activates NF-kappaB through the phosphorylation of IkappaBalpha on serines 32/36 thereby influencing the nuclear translocation of the p65 subunit. In contrast, Her-2 activates NF-kappaB independently of IkappaBalpha phosphorylation on serines 32/36. CONCLUSION: This study directly implicates ErbB receptors in the activation of NF-kappaB in PC-3 prostate cancer cells. PMID- 15880611 TI - On-line pervaporation-capillary electrophoresis for the determination of volatile acidity and free sulfur dioxide in wines. AB - Pervaporation has been coupled on-line to capillary electrophoresis (CE) by a simple interface consisting of a modified CE vial. The approach allows volatile analytes to be removed and injected into the capillary meanwhile the sample matrix remains in the pervaporator. By this approach volatile acidity and free sulfur dioxide have been simultaneously determined in wines. The detection limits (LODs) are 1.25 and 5.00 microg/mL, the quantification limits 4.12 and 16.50 microg/mL, and the linear dynamic ranges between LOD and 50 microg/mL and between 0.1 and 0.9 g/L for free sulfur dioxide and volatile acidity, respectively. The repeatability and within laboratory reproducibility, expressed as relative standard deviation (RSD), are 1.61% and 3.00% for free sulfur, and 3.35% and 4.58% for volatile acidity, respectively. The optimal pervaporation time and the time necessary for the individual separation-detection of the target analytes are 6 and 5 min, respectively. The analysis frequency is 7 h(-1) and the sample amount necessary is less than 7 mL. The proposed method and official methods for the analytes were applied to 32 wine samples. A two-tailed t-test was used to compare the methods, which yielded similar results. The errors, expressed as RSD for the two parameters, ranged between 1.3 and 4.1%. PMID- 15880612 TI - Liquid chromatography/electrospray ionization mass spectrometry for the characterization of twenty-three flavonoids in the extract of Dalbergia odorifera. AB - A method incorporating high-performance liquid chromatography (HPLC) with electrospray ionization and tandem mass spectrometry, with parallel analysis by HPLC with UV detection using a diode-array detector, was developed for the qualitative characterization of flavonoids in D. odorifera. Twenty-three flavonoids, including six isoflavones, six neoflavones, four isoflavanones, three flavanones, two chalcones, one isoflavanonol and one pterocarpan, were unambiguously identified by comparing their retention times, UV and MS spectra with those of authentic compounds. Furthermore, the collision-induced dissociations of the [M-H]- ions were studied to clarify the MS behavior of the different types of flavonoids. In negative ion ESI-MS all the flavonoids yielded prominent [M-H]- ions in the first order mass spectra. Fragments involving losses of CH3*, H2O, CO, C2H2O, and CO2 were observed in the MS/MS spectra. Each of the seven types of flavonoid showed characteristic MS/MS fragmentation patterns. The isoflavanones, flavanones and chalcones were observed to undergo retro-Diels Alder fragmentations. The spectra of almost all the neoflavonoids unexpectedly exhibited only [M-H-CH3]-* radical anions as base peaks without any further fragmentation. Substitution positions also remarkably influenced the fragmentation behavior, which could assist in distinction among the flavonoid isomers. The fragmentation rules deduced here could aid in the characterization of other flavonoids of these types. PMID- 15880613 TI - Determination of bisphenol F diglycidyl ether and related compounds by high performance liquid chromatography/mass spectrometry. PMID- 15880614 TI - Hair testing for delta9-THC-COOH by gas chromatography/tandem mass spectrometry in negative chemical ionization mode. PMID- 15880615 TI - Lobulitis is a frequent finding in prophylactically removed breast tissue from women at hereditary high risk of breast cancer. AB - The aim of this study was to investigate closely the nature of premalignant lesions that occur in prophylactically removed breast tissue from patients at hereditary high risk of breast cancer. Breast tissues obtained from 41 patients who underwent prophylactic mastectomy (pM) because of a hereditary high risk of breast cancer and control tissues from 82 age-matched healthy controls who underwent breast reduction surgery were screened for premalignant lesions. Premalignant and malignant lesions were more frequent (p = 0.0016) in pM samples (5/41) than in controls (1/82). Interestingly, lobulitis, defined as more than 100 lymphocytes and/or plasma cells per lobule in more than one section in morphologically normal lobules, was encountered in 21 of 41 (51%) pM patients, in contrast to only 8 of 82 (10%) controls (p < 0.0001). Preliminary observations indicate a predominance of T-cells in these infiltrates, in agreement with the already known frequent presence of lymphocytic infiltration in hereditary ductal in situ and infiltrating ductal/medullary carcinomas. This novel finding implies an immune reaction to an as yet unidentified antigen frequently present in women at hereditary high risk of breast cancer, possibly as part of an early carcinogenic event. PMID- 15880616 TI - In vivo effects of the fungicide ethaboxam on microtubule integrity in Phytophthora infestans. AB - The effects of ethaboxam (LGC-30473), a novel thiazole carboxamide, on microtubule organization was investigated in Phytophthora infestans (Mont) de Bary (Oomycota) at the light microscope level using standard immunofluorescence and laser scanning confocal microscopy. Hyphal cells were exposed to various concentrations ranging from 0.01 to 1.0 microg ml(-1) of ethaboxam at incubation times of 30, 60 and 120 min. Microtubules were disrupted after exposure to ethaboxam for 30 min at 0.01 microg ml(-1). Similar experimental conditions were used to document the effects of ethaboxam on microtubule integrity in the model fungus Aspergillus nidulans (Eidam) Winter (Ascomycota) and in mammalian mouse 17 clone 1 cells. Under these conditions, there was no evidence to suggest that microtubules in A. nidulans and mouse culture cells were affected by the ethaboxam. These results indicate that ethaboxam is specific for the targeting of microtubule disruption in P. infestans. PMID- 15880617 TI - Technical considerations for the use of 15N-DNA stable-isotope probing for functional microbial activity in soils. AB - Stable-isotope DNA probing is a culture-independent technique that may provide a link between function and phylogeny of active microorganisms. The technique has been used in association with 13C substrates while here we evaluate feasibility and limitations of 15N-DNA stable-isotope probing (SIP) using labelled and unlabelled pure microbial cultures or soil extracts. Our results showed that (15)N-DNA probing is feasible for cultures as well as soil samples. Limitations of 15N-DNA-SIP are (a) the need for relatively large quantities of DNA to visualise bands (although molecular resolution is much higher) and (b) 15N-DNA enrichment needed to ideally be >50 at%; however, this requirement can be lowered to approx. 40 atom% 15N with pure cultures using a modified CsCl centrifugation method (140K g for 69 h). These advances in 15N-DNA-SIP methodology open new opportunities to trace active microbial populations utilising specific N substrates in situ. PMID- 15880618 TI - Characterization of phenolic compounds in the Chinese herbal drug Tu-Si-Zi by liquid chromatography coupled to electrospray ionization mass spectrometry. AB - Phenolic compounds are the major bioactive constituents of the Chinese herbal drug Tu-Si-Zi, which is prepared from the seeds of Cuscuta chinensis. However, seeds of C. australis also are offered under the name of this drug in the herb market. In order to make a comparison of their chemical constituents, the phenolic compounds of these two Cuscuta species were analyzed by high-performance liquid chromatography/diode-array detection/electrospray ion trap tandem mass spectrometry (HPLC/DAD/ESI-MS(n)). A total of 50 compounds were observed in the methanol extracts, including 23 flavonoids, 20 lignans and 7 quinic acid derivatives. These compounds were separated on a C18 column and identified or tentatively characterized based on UV spectra and MS fragmentation behavior. In contrast to previous reports, the phenolic patterns of these two Cuscuta species were found to be very different. Kaempferol and astragalin were the predominant constituents of C. australis, while hyperoside was the major compound in C. chinensis. Most of the identified compounds, especially the acylated flavonoid glycosides, have not previously been reported from Cuscuta species. In addition, a 30 Da neutral loss observed for flavonols was investigated and could be used to differentiate flavonoid isomers such as kaempferol and luteolin. The ESI-MS fragmentation behavior of furofuran lignans was also investigated, and a characteristic pathway is proposed. The large differences observed between the phenolic constituents of C. chinensis and C. australis strongly encouraged further comparison of the bioactivities of these two species. PMID- 15880619 TI - Lentiviral transduction of human hematopoietic cells by HIV-1- and SIV-based vectors containing a bicistronic cassette driven by various internal promoters. AB - BACKGROUND: Lentiviral gene transfer into hematopoietic cells has been mostly optimized with vectors carrying a single reporter gene. For many clinical applications, lentiviral vectors should contain more than one gene because transduced cells should be enriched by a selectable marker or killed for safety reasons after use. Thus, we compared various vectors containing a bicistronic cassette driven by different ubiquitous promoters for their ability to transduce human T-lymphocytes, CD34+-cells, and dendritic cells (DCs) derived from CD34+ cells or monocytes. METHODS: We designed HIV or SIV constructs containing a bicistronic cassette composed of two reporter genes (thy1/GFP) linked by an internal ribosome entry site sequence and driven by the cytomegalovirus (CMV) or elongation factor 1alpha (EF1alpha) promoters. The woodchuck hepatitis virus post transcriptional regulatory element (WPRE) was or not inserted within the constructs, the Vpx accessory protein was or not used for SIV vectors. Target cells were infected at the same multiplicity of infection, transduction efficiency was analyzed both by flow cytometry and vector integration. RESULTS: For T-cells, HIV-based vectors/WPRE+ in which the thy1/GFP cassette was driven by the EF1alpha promoter were more efficient than SIV-based vectors. For CD34+-cells and CD34+-derived DCs, better thy1/GFP expression was achieved when the CMV promoter drove the cassette inserted into HIV-based vectors/WPRE+. Conversely, for monocyte-derived DCs, the cassette yielded better thy1/GFP expression when inserted into SIV-based vectors/WPRE+ and driven by the CMV or EF1alpha promoters, the use of Vpx significantly improving the expression levels. CONCLUSIONS: Our results provide guidelines for improving the transduction of T cells, CD34+-cells or DCs with lentiviral bicistronic vectors designed for clinical applications. PMID- 15880621 TI - A novel approach to identify plant parasitic nematodes using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Plant parasitic nematodes are difficult to identify because different species are morphologically similar, and this makes their control more difficult. The aim of this work was to develop a rapid, simple method to identify plant parasitic nematodes, based on analysis of protein profiles of nematodes generated by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOFMS). Two methods have been used: grinding and direct analysis of intact nematodes. Both methods were standardised using the nematode Anguina tritici (wheat seed-gall nematode) as a model. Development of the approach involved optimisation of experimental parameters to generate reproducible diagnostic protein profiles for plant parasitic nematodes. With alpha-cyano-4 hydroxycinnamic acid (CHCA) as the matrix, the most effective solvent extraction was with 90% acetone. With sinapinic acid (SA) as matrix, 90% ethanol was most effective. When intact nematodes were analysed directly by mixing with the matrix solution, 40 min extraction with CHCA matrix solution generated the best protein profiles. The standardised methods were applied to analyse the seed-gall nematodes A. tritici and A. funesta and to the root-knot nematode, Meloidogyne javanica, which infects many horticultural crops. Typical protein profiles and diagnostic peaks were identified for these nematode species and for mixtures of Anguina species. The results provide 'proof-of-concept' that these nematode species can be identified by protein profiling using MALDI-TOFMS. This new approach could be extended to identify other plant and non-plant parasitic nematodes. PMID- 15880622 TI - Dual linear ion trap/orthogonal acceleration time-of-flight mass spectrometer with improved precursor ion selectivity. AB - A new hybrid mass spectrometer based on dual linear ion traps (LITs) and an orthogonal acceleration time-of-flight mass spectrometer (oaTOF), that can achieve MS(n) analysis and high-mass-accuracy detection with high sensitivity, has been developed. Dual-LIT was necessary because, in a single LIT plus oaTOF combination, the LIT pressure favorable for high precursor selectivity in MS(n) analysis (less than 1 mTorr) was far different from an optimum damping pressure (50-100 mTorr) for efficient connection to the TOF mass spectrometer. A dual-LIT solved this problem of inconsistency of the optimum pressures by using the first LIT for MS(n) analysis and the second LIT for collisional damping. This dual LIT/TOF instrument achieved high-sensitivity MS(n) analysis with high precursor ion selectivity. PMID- 15880623 TI - Peptide sequencing through N-terminal phosphonylation and electrospray ionization mass spectrometry. AB - Peptides were phosphonylated at their N-termini by reacting with ethoxyphenylphosphinate in the presence of triethylamine and tetrachloromethane under mild conditions. The phosphonylated peptides were analyzed by tandem electrospray ionization mass spectrometry. N-Terminal phosphonylation selectively increased the intensities of b(n)-type ions relative to other ion types. The resulting simplified mass spectra clearly show the sequential loss of amino acid residues from the C-termini of peptides, providing a convenient and rapid method for peptide sequencing. PMID- 15880624 TI - Protein polymer drag-tags for DNA separations by end-labeled free-solution electrophoresis. AB - We demonstrate the feasibility of end-labeled free-solution electrophoresis (ELFSE) separation of DNA using genetically engineered protein polymers as drag tags. Protein polymers are promising candidates for ELFSE drag-tags because their sequences and lengths are controllable not only to generate monodisperse polymers with high frictional drag, but also to meet other drag-tag requirements for high resolution separations by microchannel electrophoresis. A series of repetitive polypeptides was designed, expressed in Escherichia coli, and purified. By performing an end-on conjugation of the protein polymers to a fluorescently labeled DNA oligomer (22 bases) and analyzing the electrophoretic mobilities of the conjugate molecules by free-solution capillary electrophoresis (CE), effects of the size and charge of the protein polymer drag-tags were investigated. In addition, the electrophoretic behavior of bioconjugates comprising relatively long DNA fragments (108 and 208 bases) and attached to uncharged drag-tags was observed, by conjugating fluorescently labeled polymerase chain reaction (PCR) products to charge-neutral protein polymers, and analyzing via CE. We calculated the amount of friction generated by the various drag-tags, and estimated the potential read-lengths that could be obtained if these drag-tags were used for DNA sequencing in our current system. The results of these studies indicate that larger and uncharged drag-tags will have the best DNA-resolving capability for ELFSE separations, and that theoretically, up to 233 DNA bases could be sequenced using one of the protein polymer drag-tags we produced, which is electrostatically neutral with a chain length of 337 amino acids. We also show that denatured (unfolded) polypeptide chains impose much greater frictional drag per unit molecular weight than folded proteins, such as streptavidin, which has been used as a drag-tag before. PMID- 15880625 TI - Determination of picomolar concentrations of proteins using novel amino reactive chameleon labels and capillary electrophoresis laser-induced fluorescence detection. AB - Py-1 and Py-6 are novel amino-reactive fluorescent reagents. The names given to them reflect that they consist of a pyrylium group attached to small aromatic moieties. Upon reaction with a primary amine there is a large spectral shift in the reagent, rendering them effectively fluorogenic. In this study, these reagents were used to label a test protein, (human serum albumin), and the sample was analyzed by capillary electrophoresis and laser-induced fluorescence detection. Detection limits after a 60 min labeling reaction at 22 degrees C (Py 1) and 50 degrees C (Py-6) were 6.5 ng/mL (98 pM) for Py-1 and 1.2 ng/mL (18 pM) for Py-6. Separation of immunoglobulin G (IgG), human serum albumin, lipase, and myoglobin after labeling with Py-6 were performed. The method was further modified to make it amenable to automation. Unlike many other amino reactive reagents used to label protein amino groups, reaction with Py-1 and Py-6 do not alter the charge of the protein and the advantage of this with respect to electrophoretic separations is discussed. PMID- 15880626 TI - A modified protein precipitation procedure for efficient removal of albumin from serum. AB - Proteomic analysis of sera and the quest for identifying serum proteins as disease markers have often been hampered by the predominance of several highly abundant proteins including albumin and immunoglobulins. Prior albumin depletion so as to enrich for otherwise undetectable serum components is therefore a prerequisite in mining the serum proteome. In the course of evaluating several available methods and commercial kits, we have been able to refine the albumin depletion protocols and establish a modified albumin removal method using trichloroacetic acid (TCA)/acetone. Changes in major protein bands were monitored by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1-D SDS-PAGE) and used as the first screening strategy to evaluate and optimize for the precipitation experimental conditions. Our method showed better performance in efficiency, specificity, and costs in comparison with two commercially available albumin removal kits, and provides a simple pre-fractionation step for the proteomic analysis of serum biomarkers. Albumin isolated by the modified method is in the native state. Our method may offer a rapid method for purifying serum albumin in large scale. PMID- 15880627 TI - Not all negative emotions are equal: the role of emotional expression in online support groups for women with breast cancer. AB - The repression/suppression of negative emotions has long been considered detrimental for breast cancer (BC) patients, leading to poor coping, progression of symptoms, and general lower quality of life. Therapies have focused on encouraging the expression of negative emotions. While group therapies have proven to be successful for BC patients, no study has looked at the role of expressing negative emotions during the therapeutic interaction. We examined written expressed emotions by women participating in a common form of psychosocial support, Internet based bulletin boards (BBs). Fifty-two new members to BC BBs were studied. They completed measures of quality of life and depression. After 6 months the measures were again assessed and messages during that time were collected and analyzed for emotional content. For the 52 women, results showed that greater expression of anger was associated with higher quality of life and lower depression, while the expression of fear and anxiety was associated with lower quality of life and higher depression. The expression of sadness was unrelated to change scores. Our results serve to challenge the commonly held belief that the expression of all negative emotions are beneficial for BC patients. Instead, expressing specific negative emotions are beneficial, while others are not. PMID- 15880628 TI - Study of the effect of the peptide loading and solvent system in SPPS by HRMAS NMR. AB - The SPPS methodology has continuously been investigated as a valuable model to monitor the solvation properties of polymeric materials. In this connection, the present work applied HRMAS-NMR spectroscopy to examine the dynamics of an aggregating peptide sequence attached to a resin core with varying peptide loading (up to 80%) and solvent system. Low and high substituted BHAR were used for assembling the VQAAIDYING sequence and some of its minor fragments. The HRMAS NMR results were in agreement with the swelling of each resin, i.e. there was an improved resolution of resonance peaks in the better solvated conditions. Moreover, the peptide loading and the attached peptide sequence also affected the spectra. Strong peptide chain aggregation was observed mainly in highly peptide loaded resins when solvated in CDCl3. Conversely, due to the better swelling of these highly loaded resins in DMSO, improved NMR spectra were acquired in this polar aprotic solvent, thus enabling the detection of relevant sequence-dependent conformational alterations. The more prominent aggregation was displayed by the VQAAIDYING segment and not by any of its intermediary fragments and these findings were also corroborated by EPR studies of these peptide-resins labelled properly with an amino acid-type spin probe. PMID- 15880629 TI - Modifiability of abnormal isometric elbow and shoulder joint torque coupling after stroke. AB - Unlike individuals with mild stroke, individuals with severe stroke are constrained to stereotypical movement patterns attributed to abnormal coupling of shoulder abductors with elbow flexors, and shoulder adductors with elbow extensors. Whether abnormal muscle coactivation and associated joint torque patterns can be changed in this population is important to determine given that it bears on the development of effective rehabilitation interventions. Eight subjects participated in a protocol that was designed to reduce abnormal elbow/shoulder joint torque coupling by training them to generate combinations of isometric elbow and shoulder joint torques away from the constraining patterns. After training, subjects demonstrated a significant reduction in abnormal torque coupling and a subsequent significant increase in ability to generate torque patterns away from the abnormal pattern. We suggest the rapid time-course of these changes reflects a residual capacity of the central nervous system to adapt to a novel behavioral training environment. PMID- 15880630 TI - Effect of body mass index on ulnar nerve conduction velocity, ulnar neuropathy at the elbow, and carpal tunnel syndrome. AB - Both high and low body mass index (BMI) have been reported as risk factors for ulnar neuropathy at the elbow (UNE), and a high BMI as a risk factor for carpal tunnel syndrome (CTS). To determine whether the extremes of BMI are risk factors for UNE or CTS, and whether BMI affects calculation of median and ulnar motor nerve conduction velocity (NCV), we retrospectively analyzed the electrodiagnostic records of control patients, UNE patients, and CTS patients. The BMI was calculated for 50 patients with a sole diagnosis of UNE and compared to the BMI of 50 patients with CTS and 50 control subjects. The mean BMIs were 25.9 +/- 4.4, 30.1 +/- 5.5, and 28.3 +/- 5.6 for the UNE, CTS, and controls, respectively. By one-way analysis of variance, the difference in BMI between the UNE patients and the normal patients was significant (P < 0.01). In the control groups, increasing BMI directly correlated with increasing ulnar motor NCV across the elbow but not with forearm NCV. Across-elbow (AE) ulnar motor NCV may be falsely increased in patients with a high BMI, probably due to distance measurement factors. Not only do relatively slender individuals have comparatively slower AE ulnar NCVs, they are also at increased risk for developing UNE. Patients with a high BMI are at increased risk for CTS. PMID- 15880631 TI - Comparison of protein and peptide prefractionation methods for the shotgun proteomic analysis of Synechocystis sp. PCC 6803. AB - Proteome analysis by gel-free "shotgun" proteomics relies on the simplification of a peptide mixture before it is analyzed in a mass spectrometer. While separation on a reverse-phase (RP) liquid chromatographic column is widely employed, a variety of other methods have been used to fractionate both proteins and peptides before this step. We compared six different protein and peptide fractionation workflows, using Synechocystis sp. PCC 6803, a useful model cyanobacterium for potential exploitation to improve its production of hydrogen and other secondary metabolites. Pre-digestion protein separation was performed by strip-based isoelectric focusing, one-dimensional polyacrylamide gel electrophoresis, or weak anion exchange chromatography, while pre-RP peptide separation was accomplished by isoelectric focusing (IEF) or strong cation exchange chromatography. Peptides were identified using electrospray ionization quadrupole time of flight-tandem mass spectrometry. Mass spectrometry (MS) and tandem mass spectra were analyzed using ProID software employing both a single organism database and the entire NCBI non-redundant database, and a total of 776 proteins were identified using a stringent set of selection criteria. Method comparisons were made on the basis of the results obtained (number and types of proteins identified), as well as ease of use and other practical aspects. IEF-IEF protein and peptide fractionation prior to RP gave the best overall performance. PMID- 15880632 TI - Methods to adjust for the interference of N2O on delta13C and delta18O measurements of CO2 from soil mineralization. AB - In this paper we present an overview of the present knowledge relating to methods that avoid interference of N2O on delta13C and delta18O measurements of CO2. The main focus of research to date has been on atmospheric samples. However, N2O is predominantly generated by soil processes. Isotope analyses related to soil trace gas emissions are often performed with continuous flow isotope ratio mass spectrometers, which do not necessarily have the high precision needed for atmospheric research. However, it was shown by using laboratory and field samples that a correction to obtain reliable delta13C and delta18O values is also required for a commercial continuous flow isotope ratio mass spectrometer. The capillary gas chromatography column of the original equipment was changed to a packed Porapak Q column. This adaptation resulted in an improved accuracy and precision of delta13C (standard deviation(Ghent): from 0.2 to 0.08 per thousand; standard deviation(Lincoln): from 0.2 to 0.13 per thousand) of CO2 for N2O/CO2 ratios up to 0.1. For delta18O there was an improvement for the standard deviation measured at Ghent University (0.13 to 0.08 per thousand) but not for the measurements at Lincoln University (0.08 to 0.23 per thousand). The benefits of using the packed Porapak Q column compared with the theoretical correction method meant that samples were not limited to small N(2)O concentrations, they did not require an extra N2O concentration measurement, and measurements were independent of the variable isotopic composition of N2O from soil. PMID- 15880633 TI - Temperature-dependent shift from labile to recalcitrant carbon sources of arctic heterotrophs. AB - Soils of high latitudes store approximately one-third of the global soil carbon pool. Decomposition of soil organic matter (SOM) is expected to increase in response to global warming, which is most pronounced in northern latitudes. It is, however, unclear if microorganisms are able to utilize more stable, recalcitrant C pools, when labile soil carbon pools will be depleted due to increasing temperatures. Here we report on an incubation experiment with intact soil cores of a frost-boil tundra ecosystem at three different temperatures (2 degrees C, 12 degrees C and 24 degrees C). In order to assess which fractions of the SOM are available for decomposition at various temperatures, we analyzed the isotopic signature of respired CO2 and of different SOM fractions. The delta13C values of CO2 respired were negatively correlated with temperature, indicating the utilization of SOM fractions that were depleted in 13C at higher temperatures. Chemical fractionation of SOM showed that the water-soluble fraction (presumably the most easily available substrates for microbial respiration) was most enriched in 13C, while the acid-insoluble pool (recalcitrant substrates) was most depleted in 13C. Our results therefore suggest that, at higher temperatures, recalcitrant compounds are preferentially respired by arctic microbes. When the isotopic signatures of respired CO2 of soils which had been incubated at 24 degrees C were measured at 12 degrees C, the delta13C values shifted to values found in soils incubated at 12 degrees C, indicating the reversible use of more easily available substrates. Analysis of phospholipid fatty acid profiles showed significant differences in microbial community structure at various incubation temperatures indicating that microorganisms with preference for more recalcitrant compounds establish as temperatures increase. In summary our results demonstrate that a large portion of tundra SOM is potentially mineralizable. PMID- 15880634 TI - Post-photosynthetic fractionation of stable carbon isotopes between plant organs- a widespread phenomenon. AB - Discrimination against 13C during photosynthesis is a well-characterised phenomenon. It leaves behind distinct signatures in organic matter of plants and in the atmosphere. The former is depleted in 13C, the latter is enriched during periods of preponderant photosynthetic activity of terrestrial ecosystems. The intra-annual cycle and latitudinal gradient in atmospheric 13C resulting from photosynthetic and respiratory activities of terrestrial plants have been exploited for the reconstruction of sources and sinks through deconvolution by inverse modelling. Here, we compile evidence for widespread post-photosynthetic fractionation that further modifies the isotopic signatures of individual plant organs and consequently leads to consistent differences in delta13C between plant organs. Leaves were on average 0.96 per thousand and 1.91 per thousand more depleted than roots and woody stems, respectively. This phenomenon is relevant if the isotopic signature of CO2-exchange fluxes at the ecosystem level is used for the reconstruction of individual sources and sinks. It may also modify the parameterization of inverse modelling approaches if it leads to different isotopic signatures of organic matter with different residence times within the ecosystems and to a respiratory contribution to the average difference between the isotopic composition of plant organic matter and the atmosphere. We discuss the main hypotheses that can explain the observed inter-organ differences in delta13C. PMID- 15880635 TI - Three sources of CO2 efflux from soil partitioned by 13C natural abundance in an incubation study. AB - This study describes a novel approach to separate three soil carbon (C) sources by one tracer method (here 13C natural abundance). The approach is based on the combination of C3 and C4 sources in different treatments, identical decomposition of C3 and C4 substances in soil, and subsequent calculation of their contribution to the total CO2 efflux. We used the temporal dynamics of the CO2 efflux from a C3 grassland soil amended with added C3 or C4 slurry and/or C3 or C4 sugar to estimate contributions of three separate C sources: native soil organic matter (SOM), slurry and sugar, to CO2 efflux. Soil with slurry and/or sugar was incubated under controlled conditions, and concentration and delta13C values of evolved CO2 were measured over a 2-week period. The main assumption needed for separation of three C sources in CO2 efflux, i.e. identical decomposition of applied C3 and C4 sugars in soil, was investigated and proven. The relative contribution to the CO2 efflux increased, but its duration decreased with an increased microbial availability of the C source, i.e. sugar > slurry > SOM. The microorganisms used the C sources according to their availability. The contribution of sugar to the CO2 efflux was finished after 2-4 days. Separation of three CO2 sources and comparison of CO2 from different treatments tracing the changes of SOM and slurry decomposition induced by addition of sugar were investigated. During the sugar decomposition (the first 2-4 days), the SOM decomposition strongly decreased. At the same time the contribution of slurry-C to CO2 increased. The shortcomings and limitations as well as possible future applications of the suggested method including FACE (Free Air Carbon dioxide Enrichments) and continuous labelling experiments are discussed. PMID- 15880636 TI - Gas-phase positive ion chemistry of 1-bromo-1-chloro-2,2,2-trifluoroethane (halothane) upon electron ionization within an ion trap mass spectrometer. AB - The positive ion chemistry occurring within an ion trap mass spectrometer upon electron ionization of 1-bromo-1-chloro-2,2,2-trifluoroethane, the important anaesthetic halothane, has been mapped by means of collision-induced decomposition and ion/molecule self-reaction experiments. Ionized halothane (M+*) reacts with neutral halothane to form the ionized olefin [ClBrC=CF2]+*. via HF elimination. Among the ionic fragments, [M-Br]+ and [M-F]+ react with halothane via chloride abstraction while [M-Cl]+ is unreactive under the same experimental conditions. Substituted methyl cations CHFX+ and CF2X+ (X = F, Cl, Br) undergo halide transfer processes, their reactivity being highest for X = F. Ionized carbenes CXY+ (X,Y = F,F; H,Br; H,Cl; H,F) react with halothane to form CClXY+ and CBrXY+, whereas CF+ inserts into the C-Cl bond to form CF3+ and CClF2+. Finally, Br+ and Cl+ react with halothane by charge transfer. Collision-induced dissociation experiments disclosed interesting rearrangements involved in the dissociations of +CHX-CF3 ions (X = Br, Cl), which undergo fluorine migration and elimination of CF2, as already observed for +CCl2-CF3 in a previous investigation. PMID- 15880637 TI - delta34S measurements on organic materials by continuous flow isotope ratio mass spectrometry. AB - Sulfur (S) isotope ratios of thoroughly dried organic samples were measured by direct thermal decomposition in an elemental analyzer coupled to an isotope ratio mass spectrometer in continuous flow mode (EA-CF-IRMS). For organic samples of up to 13 mg weight and with total S contents of more than 10 microg, the reproducibility of the delta34S(organic) values was +/-0.4 per thousand or better. However, the delta34S values of organic samples measured directly by online EA-CF-IRMS analysis were between 0.3 and 2.9 per thousand higher than those determined on BaSO4 precipitates produced by Parr Bomb oxidation from the same sample material. Our results suggest that structural oxygen in organic samples influences the oxygen isotope ratios of the SO2 produced from organic samples. Consequently, SO2 generated from organic samples appears to have different 18O/16O ratios than SO2 generated from BaSO4 precipitates and inorganic reference materials, resulting in a deviation from the true delta34S values because of 32S16O18O contributions to mass 66. It was shown that both the amount of structural oxygen in the organic sample, and the difference of the oxygen isotope ratios between organic samples and tank O2, influenced the magnitude of the observed deviation from the true delta34S value after direct EA-CF-IRMS analysis of organic samples. Suggestions are made to correct the difference between measured delta34S(organic) and true delta34S values in order to obtain not only reproducible, but also accurate S isotope ratios for organic materials by EA-CF-IRMS. PMID- 15880638 TI - Coping among older-adult, long-term cancer survivors. AB - This research focuses on the coping behavior of older adult, long-term cancer survivors. Specifically, it identifies the personal (including race and gender) and illness/treatment characteristics of survivors that are significantly associated with the use of specific coping styles: planning, acceptance, venting, denial and seeking social support. It also examines the mediating role that these forms of coping play in terms of psychological distress: anxiety, depression and cancer-related worries. Multivariate analysis of data from a random sample of 321 long-term survivors in a major cancer center tumor registry is used to address the above issues. The most prominent forms of coping used by long-term survivors were planning and acceptance; least used were venting and denial. Increased age was associated with lower use of all forms of coping, but cancer type was not. Denial as a form of coping was associated with higher anxiety, depression and cancer-related worries. While race was not found to be a significant predictor of coping style, it was a significant predictor of three dimensions of psychological distress, anxiety, depression and cancer-related worries, with African Americans exhibiting lower levels of distress than Whites. The relevance of these findings for health and social service practitioners is discussed. PMID- 15880639 TI - Evaluation of the needs and concerns of partners of women at high risk of developing breast/ovarian cancer. AB - This exploratory study investigates the experience of partners of women at high risk of developing breast/ovarian cancer and reports on the partners' views concerning their relationship, communication, future planning, children and childbearing, involvement in decision-making regarding screening and prophylactic measures, and information and support needs. In-depth interviews were conducted with 15 partners. Of these, seven were partners of women who were BRCA1/2 mutation carriers, five were partners of women with unknown mutation status, and three were partners of women who were non-carriers. None of the women had a previous diagnosis of breast or ovarian cancer. Partners of carriers and women with unknown mutation status were found to be more distressed than partners of non-carriers, with partners of mutation carriers reporting the most difficulties. Factors associated with better adjustment and coping for partners included dealing with this situation as a team with their wife, greater involvement in decision-making, satisfaction with their supportive roles and being optimistic. Decision-making difficulties in relation to prophylactic measures, concerns about their children possibly being at increased cancer risk, as well as the need to receive information directly from health professionals and the wish to meet other partners were also discussed. PMID- 15880640 TI - Phenotypic changes of adult porcine mesenchymal stem cells induced by prolonged passaging in culture. AB - The in vitro culture of porcine bone marrow-derived mesenchymal stem cells (MSCs) was used for the investigation of adult stem cell biology. Isolated porcine MSCs possessed the ability to proliferate extensively in an antioxidants-rich medium containing 5% fetal bovine serum (FBS). Greater than 40 serial MSC passages and 100 cell population doublings have been recorded for some MSC batches. Early and late passage MSCs were defined here as those cultures receiving less than 5 trypsin passages and more than 15 trypsin passages, respectively. Consistent with their robust ability to proliferate, both the early and late passage MSCs expressed the cell-cycle promoting enzyme p34cdc2 kinase. Late MSCs, however, exhibited certain features reminiscent of cellular aging such as actin accumulation, reduced substrate adherence, and increased activity of lysosomal acid beta-galactosidase. Early MSCs retained the multipotentiality capable of chondrogenic, osteogenic, and adipogenic differentiation upon induction in vitro. In contrast, late MSCs were only capable of adipogenic differentiation, which was greatly enhanced at the expense of the osteochondrogenic potential. Along with these changes in multipotentiality, late MSCs expressed decreased levels of the bone morphogenic protein (BMP-7) and reduced activity of alkaline phosphatase. Late MSCs also exhibited attenuated synthesis of the hematopoietic cytokines granulocyte colony-stimulating factor (G-CSF), leukemia inhibitory factor (LIF), and stem cell factor (SCF). The long-term porcine MSC culture, thus, provides a model system to study the molecular interplay between multiple MSC differentiation cascades in the context of cellular aging. PMID- 15880641 TI - Transferrin receptor co-localizes and interacts with the hemochromatosis factor (HFE) and the divalent metal transporter-1 (DMT1) in trophoblast cells. AB - Iron uptake and storage are tightly regulated to guarantee sufficient iron for essential cellular processes and to prevent the production of damaging free radicals. The placenta is the entry site for iron, which is delivered to the developing embryo. Iron is taken up by syncytiotrophoblast cells and is transported unidirectionally from mother to fetus against a concentration gradient. Several iron transporters and regulators were recently characterized, including DMT1 and ferroportin/Ireg1 that transport iron through membranes, and HFE that regulates TfR-mediated iron uptake. In this study we demonstrate that in a differentiated choriocarcinoma cell line BeWo, HFE, and TfR are localized mainly in recycling endosomes and a small percentage of these complexes is observed in late endosomes with DMT1 while in trophoblast cells, the level of TfR is significantly lower and it is detected with HFE and DMT1 mainly in late endosomes. Most interestingly, TfR and HFE, as well as TfR and DMT1 interact in placental trophoblast cells. Based on previous and these data we suggest that the level of intracellular iron may regulate both TfR expression (on the post transcriptional and the post-translational levels) and TfR trafficking/transcytosis in polarized cells. PMID- 15880642 TI - Disruption of protein-protein interactions: towards new targets for chemotherapy. AB - Protein-protein interactions play a key role in various mechanisms of cellular growth and differentiation, and in the replication of pathogen organisms in host cells. Thus, inhibition of these interactions is a promising novel approach for rational drug design against a wide number of cellular and microbial targets. In the past few years, attempts to inhibit protein-protein interactions using antibodies, peptides, and synthetic or natural small molecules have met with varying degrees of success, and these will be the focus of this review. PMID- 15880643 TI - Subtelomeric rearrangements in the mentally retarded: a comparison of detection methods. AB - In recent years, subtelomeric rearrangements, e.g., chromosome deletions or duplications too small to be detected by conventional cytogenetic analysis, have emerged as a significant cause of both idiopathic and familial mental retardation. As mental retardation is a common disorder, many patients need to be tested on a routine basis. In this review, we will discuss the different methods that have been applied in laboratories worldwide, including multiprobe fluorescence in situ hybridization (FISH), multiallelic marker analysis, multiplex amplifiable probe hybridization (MAPH), multiplex ligation-dependent probe amplification (MLPA), quantitative real-time PCR, comparative genomic hybridization (CGH), and multicolor FISH, including spectral karyotyping (SKY), subtelomeric combined binary ratio labeling FISH (S-COBRA FISH), multiplex FISH telomere integrity assay (M-TEL), telomeric multiplex FISH (TM-FISH), and primed in situ labeling (PRINS). PMID- 15880644 TI - Compound-specific delta13C analysis of individual amino sugars--a tool to quantify timing and amount of soil microbial residue stabilization. AB - There is strong scientific evidence that microbial residues such as amino sugars may be stabilized in soil. However, up to now, no investigation has been carried out to quantify both the amount and timing of such stabilization. This is primarily due to methodological constraints, because it is not possible to differentiate between stabilized (old) and recently produced (new) amino sugars when these biomarkers are conventionally analyzed, e.g. by means of gas chromatography and flame ionization detection. Therefore, the aim of the present study was to test whether compound-specific isotope analysis (delta13C) of amino sugars extracted from soil could be used to differentiate between old and new microbial residues. For this aim a method for the delta13C analysis of individual amino sugars was developed and optimized. First results of delta13C values of glucosamine, galactosamine, mannosamine, and muramic acid in soil samples from two different ecological studies are presented, clearly indicating that discrimination between soil inherent and newly formed amino sugars is possible in stable isotope labeling experiments. Our results further showed that, in the short term (within 1 month), only few amino sugars were built, thus making highly 13C-enriched substrates necessary for the quantification of new amino sugar production and for the determination of amino sugar turnover rates. PMID- 15880645 TI - Short-term dynamics of slurry-derived plant and microbial sugars in a temperate grassland soil as assessed by compound-specific delta13C analyses. AB - In view of recent discussions about climate change and the anthropogenically enhanced greenhouse effect, the aim of this study was to determine the short-term carbon (C) dynamics in a grassland soil after slurry application. It is known that, depending on cultivation practices, agro-ecosystems can act either as sources or as sinks for atmospheric CO2. C3 and C4 slurries were applied, differing in their stable C isotope signature, to be able to differentiate between native (soil-inherent) and fresh (slurry-applied) C. Samples were taken from 0-2, 2-7.5 and 7.5-15 cm soil depths from 90 days before until 4 weeks after slurry application at various intervals. We carried out compound-specific stable isotope analysis (CSIA) of plant- (arabinose and xylose) and microbial-derived sugars (fucose and rhamnose). Up to 45% of the applied slurry-derived xylose was found in the 0-2 cm soil depth within 24 h after slurry application, with this figure decreasing rapidly and then increasing again towards the end of the experiment. Therefore, during the first phase of slurry incorporation, preferentially the soluble part of slurry entered the first 2 cm of soil while, after about 2 weeks, particulate slurry-derived organic matter was incorporated into the soil. The ratio between plant- and microbial-derived sugars together with delta13C values of individual sugars in the 2-7.5 cm soil depth revealed that the dissipation of sugars from the 0-2 cm soil depth was not only due to leaching, but also was caused by microbial degradation of the fresh C because slurry did not contain significant amounts of rhamnose while the delta13C values of rhamnose became progressively enriched in 13C during the experiment. Stable isotope measurements of bulk soil previously only showed significant differences between C4 and C3 plots at 0-2 cm soil depth. The CSIA of the individual sugars was much more sensitive than bulk isotope measurements, revealing significant differences between C4 and C3 plots even at the 2-7.5 cm soil depth during the first phase of the experiment. Additionally, the dynamics of slurry-derived plant and microbial sugars could be followed specifically. PMID- 15880646 TI - Monooxygenase mediated pyrethroid detoxification in sea lice (Lepeophtheirus salmonis). AB - The role of monooxygenases in detoxification of the pyrethroids cypermethrin and deltamethrin was examined. Four strains of sea lice (Lepeophtheirus salmonis Kroyer) with normal or moderately reduced sensitivity towards the pyrethroids were tested in bioassays by exposure to the pyrethroid alone and in combination with an oxygenase inhibitor, piperonyl butoxide (PBO). The normal (baseline) sensitivity was considered as the sensitivity range for the two most sensitive strains. Pre-treatment with PBO elevated the sensitivity (P < 0.01) compared with groups exposed to the pyrethroid only. A positive, but not statistically significant, correlation between the activity of haem peroxidases and the pyrethroid concentration immobilizing 50% of the parasites was demonstrated (rho = 0.500 for deltamethrin and rho = 0.310 for cypermethrin). The results indicate that cytochrome P450 monooxygenases are involved in detoxification of pyrethroids in sea lice. 14C-Deltamethrin was absorbed in a lesser amount in a group of sea lice exposed to a mixture of the compound and PBO than in a group exposed to 14C deltamethrin alone. A significant difference could be demonstrated both immediately after exposure (P < 0.01) and 24 h after exposure (P < 0.05). No significant differences were found between groups pre-treated with PBO and groups exposed to 14C-deltamethrin only. 14C-Deltamethrin was taken up mainly through the cuticle, especially the cuticle on the extremities of the ventral surface, and subsequently distributed throughout the body of the parasite. PMID- 15880647 TI - Effects of a Chenopodium-based botanical insecticide/acaricide on Orius insidiosus (Hemiptera: Anthocoridae) and Aphidius colemani (Hymenoptera: Braconidae). AB - UDA-245 is a Chenopodium-based natural insecticide. Forty-eight hours after treatment with this compound, Orius insidiosus (Say) and Aphidius colemani Viereck showed slight contact toxicity at 5 g AI liter(-1). There was no residual toxicity to A colemani. These two beneficials are currently used in commercial flower and vegetable greenhouses for the management of thrips, spider mites, aphids and small caterpillars. In contrast, abamectin and insecticidal soap were toxic by contact to both species. UDA-245 did not reduce the number of eggs laid by treated O insidiosus. The eclosion of these eggs was also not adversely affected by UDA-245. The other two insecticides, abamectin and insecticidal soap had no effect on the emergence of A colemani from treated aphid mummies. However, abamectin decreased the percentage of aphid parasitism by A colemani following a residual treatment. The LC50 for UDA-245 for the two beneficials is slightly over twice the recommended field dose. PMID- 15880648 TI - Screening for trisomy 21 in monochorionic twins by measurement of fetal nuchal translucency thickness. AB - OBJECTIVE: To determine whether in screening for trisomy 21 by measurement of fetal nuchal translucency (NT) thickness in monochorionic twin pregnancies it is preferable to use the higher, smaller or average NT. METHODS: We retrospectively examined 769 monochorionic twin pregnancies that had undergone NT screening. The selection criteria were that first, in each pregnancy both fetuses were alive at the 11 to 13 + 6-week scan and second, the fetal karyotype had been determined by prenatal invasive testing or the pregnancy outcome was known. In each pregnancy the risk for trisomy 21 was calculated by a combination of maternal age and fetal NT for crown-rump length (CRL). Three estimates of risk for each pregnancy were made using the higher, smaller and average NT and these were compared for detection and false positive rates. RESULTS: The median maternal age was 33 (range, 16-45) years, the CRL was 62 (range, 45-84) mm and gestational age was 12 (range, 11 to 13 + 6) weeks. Either the fetal karyotype was normal, or phenotypically normal babies were born, in 761 cases. The karyotype was abnormal in eight cases, including six with trisomy 21. The estimated risk using the higher, smaller and average NT was 1 in 300 or more in 6 (100%), 4 (66.7%) and 6 (100%) of the trisomy 21 pregnancies and in 148 (19.4%), 57 (7.5%) and 106 (13.9%) of the normal pregnancies. For a detection rate of 100%, the false positive rates using the higher, smaller and average NT would be 5.1%, 45.9% and 4.2%, respectively. CONCLUSION: In monochorionic twins, effective screening for trisomy 21 is best provided by using the average NT measured in the two fetuses. PMID- 15880649 TI - Randomized trial of traditional dissection with electrocautery versus ultrasonic fundus-first dissection in patients undergoing laparoscopic cholecystectomy. AB - BACKGROUND: In laparoscopic cholecystectomy dissection can be with monopolar electrocautery or with ultrasonic shears, and can start at the triangle of Calot or at the fundus of the gallbladder. METHODS: Thirty-seven patients undergoing laparoscopic cholecystectomy were randomized to electrocautery dissection from the triangle of Calot and 43 to fundus-first dissection with ultrasonic shears. All procedures were strictly standardized, and patients and their postoperative carers were blinded to the operation performed. RESULTS: Ultrasonic fundus-first dissection was associated with a shorter duration of operation (mean 46 versus 61 min), fewer overnight hospital stays (two versus eight), lower pain scores 4 and 24 h after surgery, less nausea at 2, 4 and 24 h, and a shorter period of sick leave (mean 5.5 versus 9.3 days) compared with electrocautery from the triangle of Calot. CONCLUSION: Ultrasonic fundus-first dissection during laparoscopic cholecystectomy was quicker and associated with less nausea and pain than electrocautery dissection from the triangle of Calot. PMID- 15880650 TI - Stereoselective antibodies to free alpha-hydroxy acids. AB - This work describes antibodies exhibiting high stereoselectivity and class specificity towards the enantiomers of free alpha-hydroxy acids. Since the antibodies interact primarily with the carboxyl-hydroxyl-hydrogen triad about the stereogenic center, they are useful for enantiomer analysis of a variety of structurally different alpha-hydroxy acids including aromatic and aliphatic compounds, e.g. lactic acid. The utility of such antibodies for enantiomer separation in chromatography was demonstrated. Comparative studies of these and previously described anti-alpha-amino acid antibodies revealed that both types of antibodies bind only to analytes that possess both the corresponding target structure and the correct configuration. Thus, substitution of an amino group for the alpha-hydroxyl group results in a complete loss of binding activity with the anti-alpha-hydroxy acid antibodies, while an alpha-amino group is essential for the interaction between analytes and anti-alpha-amino acid antibodies. PMID- 15880651 TI - Specific requirements of sonic hedgehog signaling during oligodendrocyte development. AB - Oligodendrocyte precursors (OLPs) in the developing spinal cord are generated from the same part of the ventral neuroepithelium as motor neurons, by inductive processes that include a temporal switch from neuronal to glial cell fate. Recent studies have implicated Shh as a key signal in the generation of both OLPs and ventral neurons. In this study, we used Shh(-/-), Gli3(-/-), and Shh(-/-);Gli3(-/ ) mutants to address the role of Shh signaling during oligodendrocyte development. We find that, in the absence of Gli3, Shh signaling is dispensable for the generation and maintenance of OLPs. However, Shh is required for OLPs to emerge at the appropriate developmental stages and for subsequent differentiation of OLPs into mature oligodendrocytes. The initial delay and reduction in OLP generation in Shh(-/-);Gli3(-/-) mutants are accompanied by extended neurogenesis and persistent expression of Neurogenin 2 in the Olig2 progenitor domain, suggesting that Shh signaling influences the timing of neuron-glia fate switching. Thus, our studies suggest that Shh signaling plays multiple roles during development of oligodendrocytes. PMID- 15880652 TI - Mdm2 and HIF-1alpha interaction in tumor cells during hypoxia. AB - The interaction between HIF-1alpha, Mdm2, and p53 proteins during hypoxia has received recent attention. Here, we investigated the consequences of interaction between HIF-1alpha and Mdm2 under hypoxic conditions. Endogenous HIF-1alpha and Mdm2 proteins were co-immunoprecipitated from lysates of hypoxic HCT116 p53WT and p53(-/-) cells, suggesting that association of these two proteins is a p53 independent event. The cellular Mdm2 protein content was not significantly altered in hypoxic tumor cells. Overexpression of Mdm2 resulted in an increase in HIF-1alpha protein content in hypoxic cells and increased hypoxia-induced vascular endothelial growth factor (VEGF) transcriptional activation. These results point toward a novel and p53-independent function of Mdm2 to promote tumor cell adaptations to hypoxia by interacting with and promoting HIF-1 activation. PMID- 15880653 TI - An LRRK2 mutation as a cause for the parkinsonism in the original PARK8 family. AB - We detected a missense mutation in the kinase domain of the LRRK2 gene in members with autosomal dominant Parkinson's disease of the Japanese family (the Sagamihara family) who served as the basis for the original defining of the PARK8 Parkinson's disease locus. The results of the Sagamihara family, in combination with the unique pathological features characterized by pure nigral degeneration without Lewy bodies, provided us with valuable information for elucidating the protein structure-pathogenesis relationship for the gene product of LRRK2. We did not detect this mutation or other known mutations of the LRRK2 gene in Japanese patients with sporadic Parkinson's disease. PMID- 15880654 TI - Loss of E-cadherin mediated cell-cell adhesion as an early trigger of apoptosis induced by photodynamic treatment. AB - Photodynamic treatment with different photosensitizers (PSs) can result in the specific induction of apoptosis in many cell types. It is commonly accepted that this apoptotic response depends on the mitochondrial accumulation of the PS. Accumulation in other cellular organelles, such as lysosomes or the Golgi complex, and subsequent photodamage resulting in an apoptotic process has been also described. However, the role played by cell adhesion in apoptosis induced in epithelial cells after photodynamic treatment is not well characterized. Here, we have used a murine keratinocyte line, showing a strong dependence on E-cadherin for cell-cell adhesion and survival, to analyze the relevance of this adhesion complex in the context of zinc(II)-phthalocyanine (ZnPc) photodynamic treatment. We report that under apoptotic conditions, ZnPc phototreatment induces a rapid disorganization of the E-cadherin mediated cell-cell adhesion, which largely preceded both the detachment of cells from the substrate, via beta-1 integrins and the induction of apoptotic mitochondrial markers. Therefore, the alteration in E-cadherin, alpha- and beta-catenins adhesion proteins preceded the release of cytochrome c (cyt c) from mitochondria to the cytosol and the activation of caspase 3. In addition, blocking E-cadherin function with a specific antibody (Decma-1) induced apoptosis in this cell system. These results strongly suggest that the E-cadherin adhesion complex could be the primary target of ZnPc phototreatment, and that loss of E-cadherin mediated cell adhesion after early photodamage triggers an apoptotic response. PMID- 15880655 TI - Characterization of oligosaccharide moieties of glycopeptides by microwave assisted partial acid hydrolysis and mass spectrometry. AB - Microwave-assisted partial acid hydrolysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were used to study oligosaccharide structures of glycopeptides. Tryptic N-glycosylated peptides of horseradish peroxidase, with MH+ ions at m/z 2533, 2612, 3355, 3673, and 5647, were used as test cases. Within a microwave exposure with trifluoroacetic acid of 2 min, partial cleavages of the oligosaccharides of these tryptic N-glycosylated peptides were observed. The data showed that the most labile group within the oligosaccharides is the fucose (Fuc) residue, and that a majority of the end cleavage products are peptides with one N-acetylglucosamine (GlcNAc) residue linked to asparagine (Asn). In addition, the glycopeptides with m/z 3355 and 3673 carry an oligosaccharide (Xyl)Man3(Fuc)GlcNAc2, the glycopeptide at m/z 5647 carries two oligosaccharides (Xyl)Man3(Fuc)GlcNAc2, and the glycopeptides at m/z 2612 and 2533 carry (Xyl)Man3GlcNAc2 and (Fuc)GlcNAc, respectively. However, the glycosylation site of the m/z 2612 peptide at Asn286 is partially occupied. This simple and rapid method is particularly useful in identifying glycopeptides and obtaining monosaccharide compositions of glycopeptides. PMID- 15880656 TI - Comparison of inductively coupled plasma mass spectrometry techniques in the determination of platinum in urine: quadrupole vs. sector field. AB - In recent years the increasing use of platinum (Pt) both in medical and in industrial applications has caused its growing anthropogenic emission and spread in the environment. Pt is released into the atmosphere by exhaust catalytic converters, and Pt compounds are often used in antitumour therapies. As a consequence, significant amounts of Pt can be detected in hospital wastewaters. This can lead to an increase in the exposure levels to Pt, especially in urban areas. It is therefore necessary to determine Pt reference values in the general population, by using suitable procedures able to achieve adequate analytical performances. Several measurements of Pt in biological fluids have been reported, but the analytical methods used for the determination of Pt often lack information about the uncertainty of the results, especially for low concentrations of urinary Pt in non-occupationally exposed subjects. The present paper considers the measurement of urinary Pt levels in a general population group from central Italy, by both quadrupole (Q) and sector field (SF) inductively coupled plasma mass spectrometry (ICP-MS). The two procedures were validated and their expanded uncertainties were evaluated. The limits of detection (LODs), calculated taking into account dilution factors, were 0.18 and 0.05 ng L(-1) of Pt for the Q and SF procedures, respectively. The median value observed was 4.13 ng L(-1) of Pt in urine, while the relative combined uncertainty at 5 ng L(-1) was below 20% with both ICP-MS techniques. These data are in good agreement with those reported in the literature for similar studies. PMID- 15880657 TI - Photocatalytic degradation of the herbicide isoproturon: characterisation of by products by liquid chromatography with electrospray ionisation tandem mass spectrometry. AB - By-products arising from immobilised TiO2-catalysed photodegradation of the herbicide isoproturon [3-(4-isopropylphenyl)-1,1-dimethylurea] in aqueous solution under solar radiation were analysed by reversed-phase liquid chromatography combined with electrospray ionisation ion trap mass spectrometry. Structural information on by-products, formed at different degradation times, was then obtained from interpretation of the relevant MS/MS spectra. Several species were identified through this approach, and in many cases several isomers were found. As expected, most by-products resulted from single or multiple hydroxylation (by photo-generated OH* radicals) of the isoproturon molecule at different positions. However, substitution of some functional groups of the herbicide (isopropyl or methyl) by OH* was also observed. A possible degradation scheme is hypothesised. PMID- 15880658 TI - Changes in soil C-isotopic composition in an agroecosystem under Free Air Carbon dioxide Enrichment (FACE) treatment during a crop rotation period. AB - FACE (Free Air Carbon dioxide Enrichment) has been used since 1999 to evaluate the effects of future atmospheric CO(2) concentrations on an arable crop agroecosystem. The experiment conducted at the Institute of Agroecology at the Federal Research Centre in Braunschweig consists of a typical local crop rotation of winter barley, a cover crop, sugar beet and winter wheat. The atmospheric CO2 concentration of ambient air is about 375 ppm with a delta13C value of -7 to -9 per thousand, and 550 ppm (delta13C value = -20.2 per thousand) during daylight hours in the rings fumigated with additional CO2. Thus, the surplus C can be traced in the agricultural system. Over the course of the first experimental period (3-year crop rotation period), the C-isotopic composition and the C concentration in soil were monitored monthly. Plant samples were analysed according to the relevant developmental stages of the crop under cultivation. A 13C depletion was observed in plant parts, as well as in soil samples from the FACE rings under CO2 enrichment, indicating that labelled C has reached both respective ecosystem compartments. Albeit farming management practice (especially ploughing) leads to a mixing of 'old' and 'new' C compounds throughout all soil horizons down to the end of the ploughing layer and resulted in a heterogeneous distribution of newly formed C compounds in the soil, isotope analysis of soil C reflected where the surplus C went. PMID- 15880659 TI - Identification of novel anthrax lethal factor inhibitors generated by combinatorial Pictet-Spengler reaction followed by screening in situ. PMID- 15880660 TI - The Mini-Mental Adjustment to Cancer (Mini-MAC) scale: construct validation with a Greek sample of breast cancer patients. AB - This investigation is a Greek validation of the Mini-Mental Adjustment to Cancer (Mini-MAC) scale, an instrument derived from the MAC and designed to measure styles of coping with cancer. A sample of 225 women diagnosed with breast cancer completed this questionnaire. Statistical analyses using structural equation modeling (SEM) confirmed Watson's original five first-order factors underlying Mini-MAC items: Helplessness/Hopelessness (HH), Anxious Preoccupation (AP), Fighting Spirit (FS), Avoidance (AV) and Fatalism (F). On a higher-order level, SEM and Multidimensional Scaling revealed two second-order factors: adaptive coping (being measured by FS, AV, and F) and maladaptive coping (being measured by HH and AP). Results are discussed in terms of the theoretical framework needed to account for the relationships among Mini-MAC factors and the refinement of the applications of SEM in the study of the mental adjustment to cancer construct. PMID- 15880661 TI - Gestational sac volume measured by three-dimensional ultrasound at 11 to 13 + 6 weeks of gestation: relation to chromosomal defects. AB - OBJECTIVE: To determine the potential value of measuring the gestational sac volume (GSV) at 11 to 13 + 6 weeks of gestation in screening for chromosomal defects. METHODS: The GSV was measured using three-dimensional (3D) ultrasound in 500 consecutive singleton pregnancies immediately before chorionic villus sampling (CVS) for fetal karyotyping at 11 to 13 + 6 (median 12) weeks of gestation. RESULTS: The fetal karyotype was normal in 417 pregnancies and abnormal in 83. In the chromosomally normal group, the mean GSV increased significantly with gestational age from a mean of 69 mL at 11 weeks to 144 mL at 13 + 6 weeks (the standard deviation was 27 mL). In the chromosomally abnormal group, the mean GSV for gestational age was not significantly different from normal in fetuses with trisomy 21, trisomy 18 and Turner syndrome, but it was smaller in those with triploidy and trisomy 13. However, the mean GSV for crown rump length (CRL) was significantly larger in trisomy 18, smaller in triploidy and trisomy 13, and not different from normal in trisomy 21 and Turner syndrome. The mean CRL for gestational age was significantly smaller than normal in trisomy 18, triploidy and trisomy 13. CONCLUSIONS: The measurement of the GSV at 11 to 13 + 6 weeks of gestation is unlikely to provide useful prediction of the major chromosomal defects. In trisomy 13 and triploidy, the small GSV may be due to early onset fetal growth restriction and reduced amniotic fluid volume. In trisomy 18, the increase in GSV is probably due to the presence of associated fetal abnormalities that interfere with fetal swallowing. PMID- 15880662 TI - Automated accurate mass data processing using a gas chromatograph/time-of-flight mass spectrometer in drug discovery. AB - A gas chromatograph/time-of-flight (GCT) mass spectrometer, with high mass measurement accuracy to within 5 ppm, has been used for the automated accurate mass analysis of multicomponent mixtures and drug discovery compounds. A multicomponent mixture was analyzed several times over the course of a week to assess the reproducibility and ruggedness of the automated method while operating the GCT in electron ionization mode. For example, the data for 31 radical cations generated via electron ionization was processed using automated software (i.e. OpenLynx) to provide for mass accuracies less than 5 ppm for nearly 100% of the ions from multiple injection data. Mass accuracies of the radical anions of polyaromatic hydrocarbons generated via negative chemical ionization, and protonated pyridines and quinolines generated via methane chemical ionization, were mainly less than 5 ppm from multiple injection data. In addition, the automated method has been used for the accurate mass analysis of drug discovery compounds. PMID- 15880663 TI - Denaturant sensitive regions in creatine kinase identified by hydrogen/deuterium exchange. AB - The GdmHCl-induced unfolding of creatine kinase (CK) has been studied by hydrogen/deuterium (H/D) exchange combined with mass spectrometry. MM-CK unfolded for various periods in different denaturant concentrations was pulsed-labeled with deuterium to identify different conformational intermediate states. For all denaturation times or GdmHCl concentrations, we observed variable proportions of only two species. The low-mass envelope of isotope peaks corresponds to a species that has gained about 10 deuteriums more than native CK, and the high-mass envelope to a completely deuterated species. To localize precisely the unfolded regions in the states highly populated during denaturation, the protein was digested with two proteases (pepsin and type XIII protease) after H/D exchange and rapid quenching of the reaction. The two sets of fragments obtained were analyzed by liquid chromatography coupled to mass spectrometry to determine the deuterium level in each fragment. Bimodal distributions of deuterium were found for most peptides, indicating that these regions were either folded or unfolded. This behavior is consistent with cooperative, localized unfolding. However, we observed a monomodal distribution of deuterium in two regions (1-12 and 162-186). We conclude that the increment of mass observed in the low-mass species of the intact protein (+10 Da) has its origin in these two segments. These regions, which are very sensitive to low GdmHCl concentrations, are involved in the monomer-monomer interface of CK and their perturbation is likely to weaken the dimeric structure. At higher denaturant concentration, this would induce dissociation of the dimer. PMID- 15880664 TI - Choice of dietary protein of vegetarians and omnivores is reflected in their hair protein 13C and 15N abundance. AB - Stable isotopic (15N, 13C) composition of tissues depends on isotopic pattern of food sources. We investigated whether the isotopic compositions of human hair protein and amino acids reflect the habitual dietary protein intake. Hair samples were analyzed from 100 omnivores (selected randomly out of the 1987-1988 German nutrition survey VERA), and from 15 ovo-lacto-vegetarians (OLV), and from 6 vegans recruited separately. Hair bulk and amino acid specific isotopic compositions were analyzed by isotope-ratio mass spectrometry (EA/IRMS and GC/C/IRMS, respectively) and the results were correlated with data of the 7 day dietary records. Hair bulk 15N and 13C abundances clearly reflect the particular eating habits. Vegans can be distinguished from OLV and both are significantly distinct from omnivores in both 15N and 13C abundances. 15N and 13C abundances rose with a higher proportion of animal to total protein intake (PAPI). Individual proportions of animal protein consumption (IPAP) were calculated using isotopic abundances and a linear regression model using animal protein consumption data of vegans (PAPI = 0) and omnivores (mean PAPI = 0.639). IPAP values positively correlated with the intake of protein, meat, meat products, and animal protein. Distinct patterns for hair amino acid specific 15N and 13C abundances were measured but with lower resolution between food preference groups compared with bulk values. In conclusion, hair 13C and 15N values both reflected the extent of animal protein consumption. Bulk isotopic abundance of hair can be tested for future use in the validation of dietary assessment methods. PMID- 15880665 TI - Rapid screening of drugs of abuse and their metabolites by gas chromatography/mass spectrometry: application to urinalysis. AB - This paper describes a rapid gas chromatographic/mass spectrometric (GC/MS) screening method for the detection of drugs of abuse and/or their metabolites in urine. Synthetic stimulants, opiates, cocaine metabolites, cannabinoids--and specifically the acid metabolite of tetrahydrocannabinol (THC-COOH)--can be simultaneously extracted by a single liquid/liquid separation step, at alkaline pH, and assayed as trimethylsilyl derivatives by GC/MS in SIM (selected ion monitoring) mode. All the analytes show a good linearity (R2 > 0.99 for most of the considered substances) in the range 25-1000 ng/mL, with a good reproducibility of both the retention times (CV% <0.7) and the relative abundances of the characteristic diagnostic ions (CV% <13). The limit of detection (LOD) of the method is 25 ng/mL of target compound in human urine for most of the substances investigated, 3 ng/mL for THC-COOH, and 10 ng/mL for norbuprenorphine. Validation of the method allows its application to different fields of forensic analytical toxicology, including antidoping analysis. PMID- 15880666 TI - An application of electrospray ionization tandem mass spectrometry to probe the interaction of Ca2+/Mg2+/Zn2+ and Cl- with gramicidin A. AB - The peptide, gramicidin A (GrA), has been demonstrated to interact with divalent salts (CaCl2, MgCl2, and ZnCl2) using electrospray ionization mass spectrometry (ESI-MS). The ESI-MS analysis revealed different complexes formed due to the interaction of Val-GrA and Ile-GrA with divalent salts: [Val or Ile-GrA-H+M]+, [Val or Ile-GrA+MCl]+ and [Val or Ile-GrA+M]2+, where M is Ca or Mg or Zn. All these complexes have been subjected to collisionally activated dissociation (CAD). CAD of singly and doubly charged GrA and metal complexes exhibited the losses of water molecules, indicating the ligand preference of GrA. MS/MS and MS3 of [Val or Ile-GrA+MCl]+ resulted in the elimination of chloride ion and water, respectively. The tandem mass spectrometry data of the complex [Val-GrA+MCl]+ suggest that chloride interaction is stronger in the presence of Ca than of Mg and Zn. This study reveals that GrA could interact with Ca, Mg, and Zn in metal ion form as well as in ion pair (MCl) form. The interactions of GrA with Ca support the proposal of a physical basis for the messenger role of Ca (Urry et al., J. Biol. Chem. 1982, 257: 6659-6661). PMID- 15880667 TI - Rapid determination of urinary globotriaosylceramide isoform profiles by electrospray ionization mass spectrometry using stearoyl-d35 globotriaosylceramide as internal standard. AB - Globotriaosylceramide is a neutral glycolipid containing the trihexoside Gal(alpha1-4)Gal(ss1-4)Glc(ss1-1') covalently bound to N-acylsphingosine. It was identified as the main storage substance in the kidney of patients with Fabry disease, an X-linked deficiency of lysosomal alpha-galactosidase A which can significantly be ameliorated by enzyme replacement therapy. Unlike hemizygote males, affected heterozygote females cannot be identified by enzyme assays and therefore may remain untreated. A quantitation of urinary globotriaosylceramides was proposed as an alternative method for their diagnosis. However, the required studies on physiological and pathological variations in the excretion of trihexosides so far have been prevented by a lack of suitable methods. A validated, robust and quick high-throughput method for the quantitative analysis of globotriaosylceramide isoforms using stable-isotope-dilution/internal standardization and electrospray ionization mass spectrometry (ESI-MS) was developed. An internal standard, stearoyl-d35-globotriaosylceramide, was synthesized by enzymatic coupling of d35-stearic acid to the corresponding lyso ceramidetrihexoside. Glycolipid isoforms of high purity were obtained from a 5-mL urine portion by extraction on C18 solid-phase columns and a novel washing protocol. ESI-MS analysis was performed in full and neutral loss scan modes. Urinary trihexosyl- and some of the di- and monohexosylceramide isoforms can be quantified within a single experiment. All glycolipid isoforms were above detection limit in healthy male and female subjects (n = 63). Prominent elevations of tetracosanoyl-(C24:0 plus C24:1)-globotriaosylceramides were found in urines from female (>2.5-fold above normals) or male Fabry patients (>5.8-fold above normals), but not among controls. Globotriaosylceramide isoforms shall now be analyzed in Fabry patients, non-genetic kidney disease and healthy subjects to define the conditions for a safe diagnosis of heterozygotes. PMID- 15880668 TI - Photoionization of CH(3)I mediated by the C state in the visible and ultraviolet regions. AB - Three/two-photon resonant multiphoton ionization (MPI) of the CH3I monomer has been studied in the gas phase at 532 and 355 nm using time-of-flight mass spectrometry. Under low laser intensity (approximately 10(9) W/cm2) the mass spectra showed peaks at m/z 15, 127 and 142, corresponding to [CH3]+, [I]+ and [CH3I]+ species, at both these wavelengths. The laser power dependence for [CH3I]+, [I]+ and [CH3]+ ions showed a three-photon dependence at 532 nm. For the same three ions, photoionization studies at 355 nm gave a power dependence of 2. Both these results suggest that a vibronic energy level at approximately 7 eV, lying in the Rydberg C state, acts as a resonant intermediate level in ionization of CH3I. In the case of 355 nm, with increasing intensity additional peaks at m/z 139 and 141 were observed which could be assigned to [CI]+ and [CH2I]+ fragments. In contrast, for high intensity radiation at 532 nm ( approximately 2 x 10(10) W/cm2), only the [CI]+ fragment was observed. At these wavelengths, fragment ions observed in mass spectra mainly arise from photodissociation of the parent ion. Experiments at another wavelength in the visible region (564.2 nm) confirmed the results obtained at 532 nm. In order to assess the role of the A state in these MPI experiments, additional experiments were performed at 266 and 282.1 nm, which access the A state directly via a one-photon transition, and showed absence of a surviving precursor ion. Reaction energies for various possible dissociation channels of CH3I/[CH3I]+/[CH2I]+ were calculated theoretically at the MP2 level using the GAMESS electronic structure program. PMID- 15880669 TI - Electrospray ionization studies of transition-metal complexes of 2 acetylbenzimidazolethiosemicarbazone using collision-induced dissociation and ion molecule reactions. AB - The complexes of transition-metal ions (M2+, where M = Fe, Co, Ni, Cu, Zn, Cd, and Hg) with 2-acetylbenzimidazolethiosemicarbazone (L) are studied under electrospray ionization (ESI) conditions. The ESI mass spectra of Fe and Co complexes showed the complex ions corresponding to [M+2L-2H]+, and those of Ni and Zn complexes showed [M+2L-H]+ ions, wherein the metal/ligand ratio is 1:2 and the oxidation state of the central metal ion is +3 in the case of Fe and Co and +2 in the case of Ni and Zn. The Cd and Cu complexes showed preferentially 1:1 complex ions, i.e., [M+L-H]+ or [M+L+Cl]+, whereas Hg formed both 1:1 and 1:2 complex ions. During formation of the above complex ions one or two ligands are deprotonated after keto-enol tautomerism, depending on the nature and oxidation state of central metal ion. The structures and coordination numbers of the metal ions in the complex ions were studied by their collision-induced dissociation spectra and ion-molecule reactions with acetonitrile or propylamine in the collision cell. Based on these results it is concluded that Fe, Co, Ni and Zn form stable octahedral complexes, whereas tetrahedral or square planar complexes are formed preferentially for other metals. In addition, the Cu complex showed a [2L+2Cu-3H]+ ion with a Cu-Cu bond. PMID- 15880672 TI - TRIBUTE to Csaba Horvath, a personal recollection. PMID- 15880674 TI - Multiplex-PCR-based recombination as a novel high-fidelity method for directed evolution. AB - A new and convenient method for the in vitro recombination of single point mutations is presented. This method efficiently reduces the introduction of novel point mutations, which usually occur during recombination processes. A multiplex polymerase chain reaction (multiplex-PCR) generates gene fragments that contain preformed point mutations. These fragments are subsequently assembled into full length genes by a recombination-PCR step. The process of multiplex-PCR-based recombination (MUPREC) does not require DNase I digestion for gene-fragmentation and is therefore easy to perform, even with small amounts of target DNA. The protocol yields high frequencies of recombination without creating a wild-type background. Furthermore, the low error rate results in high-quality variant libraries of true recombinants, thereby minimizing the screening efforts and saving time and money. The MUPREC method was used in the directed evolution of a Bacillus subtilis lipase that can catalyse the enantioselective hydrolysis of a model meso-compound. Thereby, the method was proved to be useful in producing a reliable second-generation library of true recombinants from which better performing variants were identified by using a high-throughput electrospray ionization mass spectrometry (ESI-MS) screening system. PMID- 15880675 TI - Utilizing the power of microbial genetics to bridge the gap between the promise and the application of marine natural products. AB - Marine organisms are a rich source of secondary metabolites. They have yielded thousands of compounds with a broad range of biomedical applications. Thus far, samples required for preclinical and clinical studies have been obtained by collection from the wild, by mariculture, and by total chemical synthesis. However, for a number of complex marine metabolites, none of these options is feasible for either economic or environmental reasons. In order to proceed with the development of many of these promising therapeutic compounds, a reliable and renewable source must be found. Over the last twenty years, the study of microbial secondary metabolites has greatly advanced our understanding of how nature utilizes simple starting materials to yield complex small molecules. Much of this work has focused on polyketides and nonribosomal peptides, two classes of molecules that are prevalent in marine micro- and macroorganisms. The lessons learned from the study of terrestrial metabolite biosynthesis are now being applied to the marine world. As techniques for cloning and heterologous expression of biosynthetic pathways continue to improve, they may provide our greatest hope for bridging the gap between the promise and application of many marine natural products. PMID- 15880676 TI - "Parallel" and "antiparallel tail-clamps" increase the efficiency of triplex formation with structured DNA and RNA targets. AB - Sequence-specific triple-helix structures can be formed by parallel and antiparallel DNA clamps interacting with single-stranded DNA or RNA targets. Single-stranded nucleic acid molecules are known to adopt secondary structures that might interfere with intermolecular interactions. We demonstrate the correlation between a secondary structure involving the target--a stable stem predicted by in silico folding and experimentally confirmed by thermal stability and competition analyses--and an inhibitory effect on triplex formation. We overcame structural impediments by designing a new type of clamp: "tail-clamps". A combination of gel-shift, kinetic analysis, UV thermal melting and thermodynamic techniques was used to demonstrate that tail-clamps efficiently form triple helices with a structured target sequence. The performance of parallel and antiparallel tail-clamps was compared: antiparallel tail-clamps had higher binding efficiencies than parallel tail-clamps both with structured DNA and RNA targets. In addition, the reported triplex-stabilizing property of 8 aminopurine residues was confirmed for tail-clamps. Finally, we discuss the possible use of this improved triplex technology as a new tool for applications in molecular biology. PMID- 15880677 TI - Chemical synthesis of triple-labelled three-helix bundle binding proteins for specific fluorescent detection of unlabelled protein. AB - Site-specifically triple-labelled three-helix bundle affinity proteins (affibody molecules) have been produced by total chemical synthesis. The 58 aa affinity proteins were assembled on an automated peptide synthesizer, followed by manual on-resin incorporation of three different reporter groups. An orthogonal protection strategy was developed for the site-specific introduction of 5-(2 aminethylamino)-1-naphthalenesulfonic acid (EDANS) and 6-(7-nitrobenzofurazan-4 ylamino)-hexanoic acid (NBDX), constituting a donor/acceptor pair for fluorescence resonance energy transfer (FRET), and a biotin moiety, used for surface immobilization. Circular dichroism and biosensor studies of the synthetic proteins and their recombinant counterparts revealed that the synthetic proteins were folded and retained their binding specificities. The biotin-conjugated protein could be immobilized onto a streptavidin surface without loss of activity. The synthetic, doubly fluorescent-labelled affinity proteins were shown to function as fluorescent biosensors in an assay for the specific detection of unlabelled human IgG and IgA. PMID- 15880678 TI - MDA-7/IL-24 regulates proliferation, invasion and tumor cell radiosensitivity: a new cancer therapy? AB - The novel cytokine MDA-7/IL-24 was identified by subtractive hybridization in the mid-1990s as a cytokine whose expression increased during the induction of terminal differentiation, and that was either not expressed or was present at low levels in tumor cells compared to non-transformed cells. Multiple studies from several laboratories have subsequently demonstrated that expression of IL-24 in tumor cells, but not in non-transformed cells, causes their growth arrest and ultimately cell death. In addition, IL-24 has been noted to be a radiosensitizing cytokine, which in part is due to the generation of reactive oxygen species (ROS) and causing endoplasmic reticulum stress. Recent publications of Phase I trial data have shown that a recombinant adenovirus to express MDA-7/IL-24 (Ad.mda-7 (INGN 241)) was safe and had tumoricidal effects in patients, which argues that IL-24 may have therapeutic value. This review describes what is known about the impact of IL-24 on tumor cell biology in addition to approaches that may enhance the toxicity of this novel cytokine. PMID- 15880679 TI - Secalonic acid D blocks embryonic palatal mesenchymal cell-cycle by altering the activity of CDK2 and the expression of p21 and cyclin E. AB - BACKGROUND: The mycotoxin, secalonic acid D (SAD), a known animal and potential human cleft palate (CP)-inducing agent, is produced by Pencillium oxalicum in corn. SAD selectively inhibits proliferation of murine embryonic palatal mesenchymal (MEPM) cells leading to a reduction in cell numbers. These effects can explain the reduction in shelf size and the resulting CP seen in the offspring of SAD-exposed mice. Ability of SAD to inhibit proliferation as well as to block the progression of cells from G1- to S-phase of the cell-cycle were also shown in the human embryonic palatal mesenchymal (HEPM) cells suggesting the potential CP-inducing effect of SAD in human beings METHODS: Gestation day (GD) 12 mouse embryos and HEPM cells were used to test the hypothesis that the cell cycle block induced by SAD results from a disruption of stage-specific regulatory components both in vivo and in vitro. The effects of SAD on the activity of various cyclin dependent kinases (CDK) and on the levels of various positive (cyclins and CDK) and negative (CDK inhibitors p15, 16, 18, 19, 21, 27, 57) cell cycle regulators were assessed by performing kinase assays and immunoblots, respectively. RESULTS: In the murine embryonic palates, SAD specifically inhibited G1/S-phase-specific CDK2 activity, reduced the level of cyclin E and tended to increase the level of the CIP/kip CDK inhibitor, p21. In the HEPM cell cultures, exposure to IC50 of SAD significantly affected all of the above targets. In addition, a reduction in the levels/activity of CDK 4/6, a reduction in the levels of cyclins D1, D2, D3, E, A, and all INK4 family proteins, and an increase in the level of the CIP/kip CDK inhibitor, p57, were also seen. CONCLUSIONS: These results suggest that the S-phase-specific cell-cycle proteins CDK2, cyclin E and possibly p21 are the common targets of SAD in murine palatal shelves in vivo and in human embryonic palatal mesenchymal cells in vitro and may be relevant to the pathogenesis of SAD-induced CP. PMID- 15880680 TI - Two ATM variants and breast cancer risk. AB - The ATM gene is mutated in ataxia-telangiectasia (AT). Heterozygote female relatives of AT cases have a 2-7fold increased risk of breast cancer. We previously reported high risks of breast cancer associated with certain ATM variants. To estimate the risks more precisely, we have examined two ATM variants, c.1066-6T>G (IVS10-6T>G) and c.4258C>T (p.Leu1420Phe), in additional cases and controls from the same Australian cohorts previously used to estimate the risk of breast cancer associated with c.1066-6T>G. A total of 775 and 84 population-based controls were genotyped for the c.1066-6T>G and c.4258C>T ATM variants respectively, as were index cases from 378 and 373 non-BRCA1/2 breast cancer families. Penetrance was estimated by Bayes factor analysis. The allele frequencies of ATM c.1066-6T>G and c.4258C>T estimated from controls were 0.005 (95% CI=0.002 to 0.009) and 0.012 (95% CI=0.001 to 0.042), respectively. We identified three new breast cancer families with c.1066-6T>G, and seven families with c.4258C>T. Combining with the two c.1066-6T>G families previously reported, the estimated penetrance to age 70 of c.1066-6T>G was 17.2% (95% CI=4.7% to 37.5%). For c.4258C>T, the estimated average penetrance was 4.8% (95% CI 1.7% to 10.1%). In conclusion, we found no evidence that the ATM c.4258C>T variant increases breast cancer risk, and little evidence that c.1066-6T>G confers an elevated risk. Analysis of additional families will be necessary to define more precisely the risk, if any, associated with c.1066-6T>G. PMID- 15880681 TI - High frequency of ENG and ALK1/ACVRL1 mutations in German HHT patients. AB - Morbus Osler or HHT (hereditary hemorrhagic telangiectasia) is a disorder of the fibrovascular tissue that is inherited in an autosomal dominant way with frequency rates between 1:2,500 and 1:40,000. The disease provokes malformations of the blood vessels sometimes resulting in life-threatening complications. Presently, two genes involved in the development of HHT have been identified: ACVRL1 and ENG. Both of them encode proteins that belong to the TGF-beta receptor complex family and play an essential role in the formation of the vascular system. Recently, several mutations in ACVRL1 and ENG have been described in other European populations. However, no data concerning mutation frequencies in the German population have been reported so far. Therefore, we screened our collective of German HHT patients (28 single cases and 11 familial cases) for mutations in both genes by direct sequencing. We detected 11 mutations already described elsewhere and 19 novel ones. Furthermore, evidence for the pathogenic role of four new missense mutations was collected by screening a healthy control collective using RFLP analysis. Interestingly, the majority of ACVRL1 mutations represented missense mutations, whereas mutations in ENG mostly resulted in a shortened protein. Our results demonstrate the importance of ACVRL1 and ENG mutations in German HHT patients displaying mutation frequencies over 80%. PMID- 15880682 TI - Facioscapulohumeral dystrophy presenting as infantile facial diplegia and late onset limb-girdle myopathy in members of the same family. AB - We report a family with markedly variable myopathic weakness due to facioscapulohumeral muscular dystrophy (FSHD). The proband developed mild late onset proximal limb weakness. Her two daughters had severe infantile facial diplegia, initially diagnosed as Mobius syndrome, and mild childhood-onset limb weakness and scapular winging. Results of facial muscle electromyography and muscle histopathology supported a myopathic disorder. This case study further highlights the broad clinical spectrum and intrafamily variability in FSHD, and the occasional absence of a positive correlation between fragment size and disease onset. Moreover, this study underscores the importance of considering FSHD in cases of infantile facial diplegia, especially in patients not demonstrating the full clinical features of Mobius syndrome. In difficult cases, facial muscle electromyography may help to differentiate myopathic from neuropathic weakness, and help guide further diagnostic studies. PMID- 15880683 TI - Eomesodermin is expressed in mouse oocytes and pre-implantation embryos. AB - T-box genes are a highly conserved family of genes encoding transcription factors, which share a conserved DNA binding domain (the T-box). Appropriate temporal and spatial expression of this gene family is critical for gastrulation and organogenesis in a number of species. The T-box containing gene Eomesodermin was first identified in Xenopus, where it plays a critical role in mesoderm formation. In situ analyses in mice have described the expression patterns of the mouse ortholog of this gene mEomesodermin (mEomes) at the time of implantation and during fetal development. Additional studies involving the disruption of the mEomes gene, have demonstrated an additional role for mEomes in trophoblast formation. However, these analyses did not address the possibility that maternally encoded or pre-blastocyst zygotic transcription of mEomes may also contribute to embryonic development. We show here that mEomes mRNA is present prior to blastocyst formation, and that the protein product of mEomes is associated with nuclear DNA during oocyte development and persistently localizes within all nuclei of the preimplantation embryo until the early blastocyst stage. mEomes protein is associated with the meiotic spindle in the unfertilized egg and with the mitotic spindle at each cell division. Our results are consistent with mEomesodermin having a role in early preimplantation development and inner cell mass formation in addition to its function in the trophoblast lineage. PMID- 15880684 TI - Lethal and sub-lethal effects of spinosad on bumble bees (Bombus impatiens Cresson). AB - Recent developments of new families of pesticides and growing awareness of the importance of wild pollinators for crop pollination have stimulated interest in potential effects of novel pesticides on wild bees. Yet pesticide toxicity studies on wild bees remain rare, and few studies have included long-term monitoring of bumble bee colonies or testing of foraging ability after pesticide exposure. Larval bees feeding on exogenous pollen and exposed to pesticides during development may result in lethal or sub-lethal effects during the adult stage. We tested the effects of a naturally derived biopesticide, spinosad, on bumble bee (Bombus impatiens Cresson) colony health, including adult mortality, brood development, weights of emerging bees and foraging efficiency of adults that underwent larval development during exposure to spinosad. We monitored colonies from an early stage, over a 10-week period, and fed spinosad to colonies in pollen at four levels: control, 0.2, 0.8 and 8.0 mg kg(-1), during weeks 2 through 5 of the experiment. At concentrations that bees would likely encounter in pollen in the wild (0.2-0.8 mg kg(-1)) we detected minimal negative effects to bumble bee colonies. Brood and adult mortality was high at 8.0 mg kg(-1) spinosad, about twice the level that bees would be exposed to in a 'worst case' field scenario, resulting in colony death two to four weeks after initial pesticide exposure. At more realistic concentrations there were potentially important sub-lethal effects. Adult worker bees exposed to spinosad during larval development at 0.8 mg kg(-1) were slower foragers on artificial complex flower arrays than bees from low or no spinosad treated colonies. Inclusion of similar sub-lethal assays to detect effects of pesticides on pollinators would aid in development of environmentally responsible pest management strategies. PMID- 15880685 TI - Design and synthesis of hierarchical materials from ordered zeolitic building units. AB - The crystallization of colloidal silicalite-1 from clear solution is one of the best understood zeolite formation processes. Colloidal silicalite-1 formation involves a self-assembly process in which nanoslabs and nanotablets with a silicalite-1 type connectivity are formed at intermediate stages. During the assembly process, with strongly anisometric particles present, regions appear with orientational correlations, as evidenced with measurements of dynamic light scattering, viscosity, and rotation of polarized light. The presence of such regions rationalizes the unexpected differences between the crystallization kinetics under microgravity and on earth. The discovery of the locally oriented regions sheds new light on currently poorly understood hydrodynamic effects on the zeolite formation processes, such as the influence of stirring on the phases obtained and the subsequent kinetics. Addition of surfactants or polymers modifies the ordering of the zeolitic building units in the correlated regions, and new types of hierarchical materials named zeogrids and zeotiles can be obtained. PMID- 15880686 TI - Structurally diverse second-generation [2.2]paracyclophane ketimines with planar and central chirality: syntheses, structural determination, and evaluation for asymmetric catalysis. AB - A set of 20 novel [2.2]paracyclophane ketimines with planar and central chirality has been synthesized from enantiomerically pure and racemic 5-acyl-4 hydroxy[2.2]paracyclophane and alpha-branched chiral amines. Their X-ray structures were determined to elucidate the three-dimensional structures and the absolute configuration. The ketimines were used as catalysts in the asymmetric 1,2-addition reactions of diethylzinc with substituted benzaldehydes to furnish chiral alcohols in up to 95 % ee. PMID- 15880687 TI - Simple synthesis of aminoquinoline/ethylaniline copolymer semiconducting nanoparticles. AB - Pure copolymer nanoparticles from 8-aminoquinoline (AQ) and 2-ethylaniline (EA) were easily synthesized by a chemically oxidative polymerization in three different aqueous media. The potential and temperature of polymerization solution were used to successfully follow the polymerization progress. The molecular and morphological structures of the resulting AQ/EA copolymer particles were systematically characterized by IR, UV/Vis, NMR, gel permeation chromatography, laser particle-size analysis, atomic force and transmission electron microscopy. The oxidation potential of the monomers as well as the polymerization yield, structure, and properties of the particles were found to significantly depend on AQ/EA ratio, polymerization temperature and medium. It is surprisingly found that AQ homopolymerization and AQ/EA (50:50) copolymerization at 5 degrees C in HCl simply afford nano-ellipsoids with the major/minor axis diameters of 24/14 nm and 80/67 nm, respectively. A simple method of synthesizing semiconducting pure nanoparticles by introducing the AQ units with positively charged quaternary ammonium groups but in the absence of adscititious stabilizer or sulfonic substituent on the monomers is established first. Both the molecular weight and bulk electroconductivity of the copolymers exhibit a maximum at AQ content of 10 mol %. The solubility and film formability of the copolymers are good in highly polar solvents and reach the optimal at the AQ content of 20 and 10 mol %, respectively. PMID- 15880688 TI - Synthesis of multi-1,10-phenanthroline ligands with 1,3-phenylene linkers and their lithium complexes. AB - The synthesis of two multisite ligands containing four and five 1,10 phenanthroline (phen) chelates in line, respectively, is presented. The connectors are 1,3-phenylene linkers. The two ligands were prepared following multistep procedures, the two key reactions being the Suzuki coupling reaction between aromatic nuclei and the nucleophilic addition of aryllithium derivatives onto a phen fragment. The coordination chemistry of both ligands with Li+ ions was very clean and selective, whereas their reaction with copper(I) led to intractable mixtures of insoluble complexes. The tetraphen and the pentaphen compounds afforded almost quantitatively the four- and five-lithium double stranded helical complexes, respectively. The helical systems are probably highly wound, as indicated by NMR measurements. The pronounced strain of the 5-Li+ complex is reflected by the easy loss of a lithium cation, as shown by electrospray mass spectrometry. PMID- 15880689 TI - Surgical management of parapharyngeal space masses. AB - BACKGROUND: We sought to examine surgical techniques used to remove parapharyngeal space (PPS) masses. METHODS: This retrospective search was conducted from 1980 to 2003. Age, sex, diagnosis, surgical approach, complications, and outcome were collected. RESULTS: One hundred sixty-six PPS masses were identified: 21 (12.7%) were malignant, 145 (87.3%) were benign, 76 (45.8%) were vascular, and 69 (41.6 %) involved the skull base. Transcervical techniques were used in all cases. Removing the styloid and its musculature and level II lymphadenectomies enhanced exposure for vascular and skull base tumors. Thirty transcervical-transmastoid dissections (20.4%) facilitated removal of vascular skull base tumors. To identify the facial nerve, 20 transparotid transcervical approaches (13.6%) were performed. Three mandibulotomies (2.0%) were required for internal carotid artery involvement. Expected neurologic sequelae resulted from cranial nerve involvement by tumor. Three patients (2.0%), all presenting with recurrent cancer, had local recurrences. CONCLUSION: Careful patient assessment and surgical techniques allow the oncologically safe removal of benign, vascular, and skull base PPS tumors. PMID- 15880690 TI - Post-transcriptional gene regulation by gamma herpesviruses. AB - The Epstein-Barr virus (EBV) SM protein is a member of a highly conserved family of proteins present in most mammalian herpes viruses. There is a significant amount of functional and sequence divergence among the homologs encoded by the human herpes viruses, including differences in mechanism of action and varying effects on splicing and transcription. Nevertheless, in those cases where it has been studied, these proteins are essential for lytic replication of the virus. The mechanism by which SM regulates gene expression operates at the level of mRNA stability, processing, and export. SM enhances expression of EBV lytic genes and has both positive and negative effects on cellular gene expression. In addition to enhancing accumulation of EBV gene mRNAs, SM has important effects on cellular mRNAs, altering the host cell gene expression profile to facilitate viral replication. This article describes the current state of knowledge regarding the role of EBV SM in cellular and viral gene regulation and summarizes some of the similarities and differences with the ORF57 homolog from Kaposi's sarcoma associated herpes virus (KSHV/HHV8). PMID- 15880691 TI - Inhibition of extracellular signal regulated kinase (ERK) leads to apoptosis inducing factor (AIF) mediated apoptosis in epithelial breast cancer cells: the lack of effect of ERK in p53 mediated copper induced apoptosis. AB - Recent studies have shown that MEK/ERK-mediated signals play a major role in regulation of activity of p53 tumor suppressor protein. In this study, we investigated whether or not there is functional interaction between p53 and MEK/ERK pathways in epithelial breast cancer cells exposed to copper or zinc. We demonstrated that expression of wild-type p53 induced by copper or zinc significantly reduced phosphorylation of extracellular signal regulated kinase (ERK) in epithelial breast cancer MCF7 cells. Mutation or suppression of p53 in MDA-MB231 and MCF7-E6 cells, respectively, resulted in a strong ERK phosphorylation in the presence of metals. Weak ERK phosphorylation in MCF7 cells induced by copper or zinc was linked to mitochondrial disruption and apoptosis. Furthermore, inhibition of ERK through addition of PD98059 stimulated p53 activation in MCF7 cells and also led to upregulation of p53 downstream targets, p21 and Bax, which is a proapototic member of Bcl-2 family triggering mitochondrial pore opening. Moreover, blockage of the MEK/ERK pathway caused a breakdown of the mitochondrial membrane potential accompanied by an elevation in the ROS production. Disruption of p53 expression attenuated the depolarization of the mitochondrial membrane and ROS generation. Furthermore, PD98059 initiated apoptosis inducing factor (AIF) translocation from mitochondria to the nucleus in MCF7 cells; which are depleted in caspase 3. Interestingly, repression of MEK/ERK pathway did not intensify the cell stress caused by metal toxicity. Therefore, these findings demonstrate that MEK/ERK pathway plays an important role in downregulation of p53 and cell survival. Inhibition of ERK can lead to apoptosis via nuclear relocation of AIF. However, metal-induced activation of p53 and mitochondrial depolarization appears to be independent of ERK. Our data suggest that copper induces apoptosis through depolarization of mitochondrial membrane with release of AIF, and this process is MEK/ERK independent. PMID- 15880692 TI - Cellular and molecular interactions regulating skeletogenesis. AB - Skeletal development involves complex coordination among multiple cell types and tissues. In long bones, a cartilage template surrounded by the perichondrium is first laid down and is subsequently replaced by bone marrow and bone, during a process named endochondral ossification. Cells in the cartilage template and the surrounding perichondrium are derived from mesenchymal cells, which condense locally. In contrast, many cell types that make up mature bone and in particular the bone marrow are brought in by the vasculature. Three tissues appear to be the main players in the initiation of endochondral ossification: the cartilage, the adjacent perichondrium, and the invading vasculature. Interactions among these tissues are synchronized by a large number of secreted and intracellular factors, many of which have been identified in the past 10 years. Some of these factors primarily control cartilage differentiation, while others regulate bone formation and/or angiogenesis. Understanding how these factors operate during skeletal development through the analyses of genetically altered mice depends on being able to distinguish the effect of these molecules on the different cell types that comprise the skeleton. This review will discuss the complexity of skeletal phenotypes, which arises from the tightly regulated, complex interactions among the three tissues involved in bone development. Specific examples illustrate how gene functions may be further assessed using new approaches including genetic and tissue manipulations. PMID- 15880693 TI - Changes in mechanical properties and cellularity during long-term culture of collagen fiber ACL reconstruction scaffolds. AB - Resorbable scaffolds for anterior cruciate ligament (ACL) reconstruction should provide temporary mechanical function then gradually breakdown while promoting matrix synthesis by local cells. Crosslinking influences collagen's mechanical properties, degradation rate, and interactions with cells. Our objective was to compare the effects of different crosslinkers on cellularity and mechanical properties during long-term (8 week) culture of collagen fiber scaffolds. Fibers were fabricated from an acid-insoluble dispersion of bovine dermal collagen and crosslinked with either ultraviolet irradiation (UV; a physical crosslinker) or 1 ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC; a chemical crosslinker). Scaffolds consisted of 50 fibers bundled in parallel. Initial attachment of fibroblasts was similar on both scaffolds; however, from 1 to 8 weeks in culture, UV-crosslinked scaffolds had significantly more cells attached than EDC crosslinked scaffolds. The initial breaking load (3.50 N) and stiffness (2.23 N/mm) of EDC-crosslinked scaffolds were significantly greater than those of UV crosslinked scaffolds (2.32 N; 1.21 N/mm) and were unaffected by long-term fibroblast culture. In contrast, the load-bearing capacity of fibroblast-seeded UV-crosslinked scaffolds decreased 60% to 0.91 N after 8 weeks in culture. EDC crosslinked scaffolds maintained strength and moderate cellularity; UV crosslinked scaffolds, in contrast, were highly cellular, but had poor mechanical properties that decreased during culture. These in vitro results suggest that collagen fiber scaffolds crosslinked with EDC may be more suitable for ACL reconstruction than those crosslinked with UV. PMID- 15880694 TI - Overexpression of regucalcin suppresses cell death and apoptosis in cloned rat hepatoma H4-II-E cells induced by insulin or insulin-like growth factor-I. AB - The role of regucalcin, a regulatory protein in intracellular signaling pathway, in cell death was investigated by using the cloned rat hepatoma H4-II-E cells overexpressing regucalcin. The hepatoma cells (wild-type) and stable regucalcin/pCXN2 transfectants were cultured for 72 h in a medium containing 10% fetal bovine serum (FBS) to obtain subconfluent monolayers. After culture for 72 h, cells were further cultured for 24-72 h in a medium containing either vehicle, insulin (10(-8) or 10(-7) M) or insulin-like growth factor-I (IGF-I; 10(-9) or 10(-8) M) in the absence of FBS. The number of wild-type cells was significantly decreased by culture for 24, 48, or 72 h in the presence of insulin (10(-8) or 10(-7) M) or IGF-I (10(-9) or 10(-8) M). Agarose gel electrophoresis showed the presence of low-molecular-weight deoxyribonucleic acid (DNA) fragments of adherent wild-type cells cultured with insulin or IGF-I. The effect of insulin or IGF-I in stimulating cell death and DNA fragmentation in hepatoma cells (wild type) was significantly prevented in transfectants overexpressing regucalcin. Meanwhile, epinephrine (10(-6) or 10(-5) M) or transforming growth factor-beta1 (10(-13) or 10(-12) M) did not cause cell death of hepatoma cells. Insulin induced decrease in the number of wild-type cells was significantly prevented by culture with caspase-3 inhibitor (10(-8) M), although the effect of IGF-I was not inhibited. The effect of insulin or IGF-I in inducing the death of hepatoma cells (wild-type) was significantly prevented in the presence of N omega-nitro-L arginine methylester (NAME), an inhibitor of nitric oxide synthase. Genistein (10(-6) M), an inhibitor of protein tyrosine kinase, or vanadate (10(-5) M), an inhibitor of protein tyrosine phosphatase, caused a significant decrease in the number of hepatoma cells (wild-type). The effect of insulin in inducing the death of wild-type cells was not seen in the presence of genistein or vanadate. The effect of IGF-I on the death of wild-type cells was observed in the presence of genistein or vanadate. The effect of genistein on cell death was significantly prevented in transfectants. Such effect was not seen with vanadate. This study demonstrates that insulin or IGF-I stimulates cell death and apoptosis in the hepatoma cells, and that overexpression of regucalcin has a suppressive effect on cell death induced by insulin or IGF-I that is mediated through different signaling pathway. PMID- 15880695 TI - Metal ions as cofactors for the binding of inhibitors to methionine aminopeptidase: a critical view of the relevance of in vitro metalloenzyme assays. PMID- 15880696 TI - Size-selective and reversible encapsulation of single small hydrocarbon molecules by a cavitand-porphyrin species. PMID- 15880697 TI - Pap smears of patients with extramammary Paget's disease of the vulva. AB - Extramammary Paget's disease (EMPD) of the vulva is a rare entity. The diagnosis is almost always made on biopsy. Tumor cells are seen rarely in Papanicolaou (Pap) smears. We encountered three cases of EMPD that were detected in Pap smears. One patient had vulvar and vaginal involvement and the abnormal cells seen in the vaginal smear initially were interpreted as high-grade squamous intraepithelial lesion. Retrospective review showed scattered single atypical cells with enlarged hyperchromatic nuclei, coarse chromatin, inconspicuous nucleoli, high nuclear/cytoplasmic (N:C) ratio, and scanty basophilic cytoplasm. Rare signet ring cells and cells within cells were present. In the other two patients who had cervical involvement, the correct diagnosis was made on Pap smears. The slides showed both single and cohesive sheets of glandular cells with enlarged round to oval nuclei, coarse chromatin, prominent nucleoli, and abundant basophilic cytoplasm containing prominent vacuoles with signet ring-cell appearance. Cells within cells were abundant. EMPD has distinct cytomorphological features. Although infrequently encountered, EMPD can be diagnosed on Pap smears with adequate clinical history. PMID- 15880698 TI - Spontaneous infarction of a pleomorphic adenoma in parotid gland: diagnostic problems and review. AB - Although infarction of parotid gland pleomorphic adenoma (PA) following fine needle aspiration (FNA) has been well-documented, spontaneous infarction of PA has remained as an uncommon entity in the literature. To our knowledge, we report the second case of spontaneous infarction occurring in a parotid gland PA. A 44 yr-old man presented with a 2-yr history of slowly enlarging right parotid mass, which had become painful 1 mo before performing FNA biopsy. Smears revealed abundant necrotic debris, atypical squamous cells, and small cells with dark nuclei suggestive of a carcinoma. Histologic examination of the tissue fragments demonstrated degenerated clusters of cells and chondromyxoid matrix. The parotidectomy specimen had features consistent with those of an infarcted PA. Although appears to be rare, spontaneous infarction of PA should be considered in the differential diagnosis, since necrosis may mimic carcinoma and cause misinterpretation of necrosis as an indication of malignancy, in an otherwise benign salivary gland neoplasm. PMID- 15880699 TI - Fine-needle aspiration biopsy of parotid cystic lesion with crystalloid formation. PMID- 15880700 TI - Teratogen update: lead and pregnancy. AB - This review focuses on the impacts of lead exposure on reproductive health and outcomes. High levels of paternal lead exposure (>40 microg/dl or >25 microg/dl for a period of years) appear to reduce fertility and to increase the risks of spontaneous abortion and reduced fetal growth (preterm delivery, low birth weight). Maternal blood lead levels of approximately 10 microg/dl have been linked to increased risks of pregnancy hypertension, spontaneous abortion, and reduced offspring neurobehavioral development. Somewhat higher maternal lead levels have been linked to reduced fetal growth. Some studies suggest a link between increased parental lead exposure and congenital malformations, although considerable uncertainty remains regarding the specific malformations and the dose-response relationships. Common methodological weaknesses of studies include potential exposure misclassifications due to the frequent unavailability of exposure biomarker measurements at biologically appropriate times and uncertainty regarding the best exposure biomarker(s) for the various outcomes. A special concern with regard to the pregnant woman is the possibility that a fetus might be exposed to lead mobilized from bone stores as a result of pregnancy-related metabolic changes, making fetal lead exposure the result of exposure to exogenous lead during pregnancy and exposure to endogenous lead accumulated by the woman prior to pregnancy. By reducing bone resorption, increased calcium intake during the second half of pregnancy might reduce the mobilization of lead from bone compartments, even at low blood lead levels. Subgroups of women who incurred substantial exposures to lead prior to pregnancy should be considered to be at increased risk. PMID- 15880701 TI - Effects of epidermal growth factor (EGF), transforming growth factor-alpha (TGFalpha), and 2,3,7,8-tetrachlorodibenzo-p-dioxin on fusion of embryonic palates in serum-free organ culture using wild-type, EGF knockout, and TGFalpha knockout mouse strains. AB - BACKGROUND: 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is teratogenic in mice, producing cleft palate (CP). TCDD exposure disrupts expression of epidermal growth factor (EGF) receptor, EGF, and transforming growth factor-alpha (TGFalpha) in the palate and affects proliferation and differentiation of medial epithelial cells. EGF knockout embryos are less susceptible to the induction of CP by TCDD. This study used palate organ culture to examine the hypothesis that EGF enables a response to TCDD. METHODS: The midfacial tissues from wild-type (WT), EGF knockout, C57BL/6J, and TGFalpha knockout embryos were placed in organ culture on gestational day (GD) 12. Palatal explants were cultured for 4 days in serum-free Bigger's (BGJ) medium with 0.1% dimethyl sulfoxide (DMSO) or 1 x 10( 8) M TCDD with or without 2 ng of EGF/ml, 1 or 2 ng of TGFalpha/ml. Effects on palatal fusion were evaluated on day 4 of culture. EGF levels in explants and medium were determined using Luminex technology. RESULTS: In serum-free, control medium, palates from all of the strains fused. EGF knockout palates cultured with TCDD (no EGF) fused, but those cultured with TCDD + 2 ng of EGF/ml failed to fuse (p < 0.05 vs. control or TCDD without EGF). TGFalpha knockout palates failed to fuse when cultured with TCDD + 2 ng of TGFalpha/ml. EGF levels increased in tissue and accumulated in the medium after 24 hr of culture. CONCLUSIONS: This study demonstrated that providing EGF to the palates of EGF knockout mice restored the response to TCDD. These studies support the hypothesis that the mechanism for induction of CP by TCDD is mediated via the EGFR pathway. PMID- 15880702 TI - A proposed mechanism for intermediate atresia ani (AA), based on a porcine case of AA and hypospadias. AB - BACKGROUND: Atresia Ani (AA) is one of the most common malformations of the lower gastrointestinal tract. It occurs in 1 in 1500 to 1 in 5000 human births. These patients suffer physically and psychologically from this disorder, and thus there is a research interest in the condition. There are 3 subcategories of AA: high, intermediate, and low. Each of these forms is likely related to separate etiological processes. METHODS: An anatomical study was performed on a porcine case of AA with a urorectal fistula and malformed urethra. RESULTS: This animal was found to have the intermediate form of AA. CONCLUSIONS: A new mechanism is hypothesized, distinct from that described for the high and low forms of AA. This proposed mechanism involves the persistence of the cloacal membrane. Evidence to support this hypothesis includes: location of the urorectal fistula in the region of the embryonic cloacal duct, the lack of anomalies usually seen in conjunction with AA associated with mutations of the Sonic Hedgehog gene, and the presence of a malformed urethra, which is significantly correlated to intermediate AA. This form of AA could be related to a failure of the cloacal membrane to break down at the appropriate time during urorectal separation. PMID- 15880703 TI - Osteogenic oxysterols inhibit the adverse effects of oxidative stress on osteogenic differentiation of marrow stromal cells. AB - The osteoporosis that occurs with aging is associated with reduced number and activity of osteoblastic cells. Aging, menopause, and osteoporosis are correlated with increased oxidative stress and reduced antioxidant defense mechanisms. We previously demonstrated that oxidative stress induced by a variety of compounds such as xanthine/xanthine oxidase (XXO) and minimally oxidized LDL (MM-LDL) inhibit the osteogenic differentiation of osteoprogenitor cells. Oxysterols are a family of products derived from cholesterol oxidation that have important biological activities. Recently, we reported that a specific oxysterol combination consisting of 22(S)- or 22(R)-hydroxycholesterol and 20(S) hydroxycholesterol has potent osteogenic properties in vitro when applied to osteoprogenitor cells including M2-10B4 (M2) marrow stromal cells. We now demonstrate that this osteogenic combination of oxysterols prevents the adverse effects of oxidative stress on differentiation of M2 cells into mature osteoblastic cells. XXO and MM-LDL inhibited the osteogenic differentiation of M2 cells, demonstrated by the inhibition of markers of osteogenic differentiation: alkaline phosphatase activity, osteocalcin expression and mineralization. Treatment of M2 cells with osteogenic oxysterol combination 22(S)- and 20(S) hydroxycholesterol both blocked and reversed the inhibition of osteogenic differentiation produced by XXO and MM-LDL in these cells. The protective effect of the oxysterols against oxidative stress was dependent on cyclooxygenase 1 and was associated with the osteogenic property of the oxysterols. These findings further demonstrate the ability of the osteogenic oxysterols to positively regulate osteogenic differentiation of cells, and suggests that the use of these compounds may be a novel strategy to prevent the adverse effects of oxidative stress on osteogenesis. PMID- 15880704 TI - Parathyroid hormone stimulation and PKA signaling of latent transforming growth factor-beta binding protein-1 (LTBP-1) mRNA expression in osteoblastic cells. AB - Parathyroid hormone (PTH) regulates bone remodeling and calcium homeostasis by acting on osteoblasts. Recently, the gene expression profile changes in the rat PTH (1-34, 10(-8)M)-treated rat osteoblastic osteosarcoma cell line, UMR 106-01, using DNA microarray analysis showed that mRNA for LTBP-1, a latent transforming growth factor (TGF-beta)-binding protein is stimulated by PTH. Latent TGF-beta binding proteins (LTBPs) are required for the proper folding and secretion of TGF beta, thus modifying the activity of TGF-beta, which is a local factor necessary for bone remodeling. We show here by real time RT-PCR that PTH-stimulated LTBP-1 mRNA expression in rat and mouse preosteoblastic cells. PTH also stimulated LTBP 1 mRNA expression in all stages of rat primary osteoblastic cells but extended expression was found in differentiating osteoblasts. PTH also stimulated TGF beta1 mRNA expression in rat primary osteoblastic cells, indicating a link between systemic and local factors for intracellular signaling in osteoblasts. An additive effect on LTBP-1 mRNA expression was found when UMR 106-01 cells were treated with PTH and TGF-beta1 together. We further examined the signaling pathways responsible for PTH-stimulated LTBP-1 and TGF-beta1 mRNA expression in UMR 106-01 cells. The PTH stimulation of LTBP-1 and TGF-beta1 mRNA expression was dependent on the PKA and the MAPK (MEK and p38 MAPK) pathways, respectively in these cells, suggesting that PTH mediates its effects on osteoblasts by several intracellular signaling pathways. Overall, we demonstrate here that PTH stimulates LTBP-1 mRNA expression in osteoblastic cells and this is PKA dependent. This event may be important for PTH action via TGF-beta in bone remodeling. PMID- 15880705 TI - Mutations of COL10A1 in Schmid metaphyseal chondrodysplasia. AB - Schmid metaphyseal chondrodysplasia (SMCD) is a dominantly inherited cartilage disorder caused by mutations in the gene for the hypertrophic cartilage extracellular matrix structural protein, collagen X (COL10A1). Thirty heterozygous mutations have been described, about equally divided into two mutation types, missense mutations, and mutations that introduce premature termination signals. The COL10A1 mutations are clustered (33/36) in the 3' region of exon 3, which codes for the C-terminal NC1 trimerization domain. The effect of COL10A1 missense mutations have been examined by in vitro expression and assembly assays and cell transfection studies, which suggest that a common consequence is the disruption of collagen X trimerization and secretion, with consequent intracellular degradation. The effect of COL10A1 nonsense mutations in cartilage tissue has been examined in two patients, demonstrating that the mutant mRNA is completely removed by nonsense mediated mRNA decay. Thus for both classes of mutations, functional haploinsufficiency is the most probable cause of the clinical phenotype in SMCD. PMID- 15880706 TI - Interspecialty and intraspecialty differences in the management of thyroid nodular disease and cancer. AB - INTRODUCTION: The management of thyroid cancer includes multiple medical specialties. Physicians from different specialties may vary in opinion regarding the optimal investigation and treatment of patients. Little data exist evaluating the differences within or between various specialties treating thyroid disease. This study aims to examine responses from a variety of specialty physicians closely involved in the medical or surgical management of thyroid disease to provide evidence as to whether any difference exists. METHODS: A cross-sectional survey of attendees at the 5(th) Biennial Course on the Management of Thyroid Nodular Disease and Cancer was conducted using an anonymous electronic touch pad system. Touch pads were given to 213 attendees who were asked to respond to 44 questions. This study analyzes the responses obtained from 19 selected questions (43%) and compares the results between endocrinologists (n = 48), general surgeons (n = 41), otolaryngologists (n = 61), and pathologists (n = 20). RESULTS: Responses were obtained from 69% of endocrinologists, 68% of general surgeons, 72% of otolaryngologists, and 65% of pathologists. Statistically significant interspecialty differences were observed in 12 (63%) of 19 questions. Each question and a summary of responses from all touch pads were recorded. CONCLUSIONS: Significant differences in the attitudes toward, and presumably the practice of, managing thyroid nodular disease and cancer exist between specialties. An understanding of these differences is helpful when working as a multidisciplinary team to optimize patient care. PMID- 15880707 TI - Confined electron-transfer reactions within a molecular metal oxide "Trojan Horse". PMID- 15880708 TI - Asymmetric carboselenenylation reaction of alkenes with aromatic compounds. PMID- 15880709 TI - The diagnosis of Trichomonas vaginalis in liquid-based Pap tests: correlation with PCR. AB - The conventional Papanicolaou smear (CPS) is not considered accurate for the diagnosis of Trichomonas vaginalis (T. vaginalis), and women noted to carry the organism on CPS are recommended to undergo confirmatory testing. Liquid-based preparations have been shown to facilitate the diagnosis of squamous lesions and may also facilitate the diagnosis of T. vaginalis. We used polymerase chain reaction (PCR) to investigate the accuracy of the diagnosis of T. vaginalis by the liquid-based Pap test (LBP). LBP with the diagnosis of T. vaginalis from a 12 mo period were identified. Residual samples from these cases were subjected to PCR for T. vaginalis as were the residual samples of a control group of 195 LBP (including 103 inflammatory LBP and 69 cases of atypical squamous cells) in which T. vaginalis was not diagnosed cytologically. PCR confirmed the presence of T. vaginalis in 50 of 51 (98%) LBP and identified 2 additional cases. Morphologic identification of T. vaginalis on LBP is highly accurate and should not require confirmatory testing. PMID- 15880710 TI - Aspiration biopsy of nodular pseudoangiomatous stromal hyperplasia of the breast: clinicopathologic correlates in 10 cases. AB - Nodular pseudoangiomatous stromal hyperplasia (PASH) of the breast is rare and often indistinguishable from fibroadenoma, clinically and on aspiration biopsy smears. We report our observations in 10 patients with PASH, evaluated by fine needle aspiration (FNA) biopsy and core biopsy. We retrospectively reviewed the clinical, radiographic, cytologic, and histologic findings in 10 cases of pure nodular PASH. Ten patients with a presumed clinical and radiologic diagnosis of fibroadenoma underwent aspiration biopsy. The aspiration smears were diagnosed as fibroadenoma (4 cases), cellular fibroadenoma (1 case), schwannoma versus neurofibroma (1 case), fibrocystic change (3 cases; 2 with atypia), and "not specific for a lesion" (1 case). A diagnosis of PASH was not suspected in any case. A discrepant or imprecise cytologic diagnosis and/or the presence of dissociated spindle or epithelial cells, or cellular stromal fragments prompted a surgical excision in 7 of 10 patients (70%). The remaining 3 patients exhibited cytologic features of fibroadenoma and were diagnosed as such; however, surgical excision was recommended. Three patients underwent a subsequent core biopsy, with a diagnosis of PASH being made in 1 patient. FNA biopsy could not discriminate PASH from fibroadenoma in 4 of 10 patients (40%) or suggest a diagnosis of PASH in any case. On retrospective review, the finding of plump, spindle-shaped mesenchymal cells may be a cytologic clue to suggest a diagnosis of PASH. PMID- 15880711 TI - Fine-needle aspiration biopsy of ectopic cervical spindle-cell thymoma: a case report. AB - In this report, we describe a case of ectopic cervical thymoma (medullary or type A thymoma, according to the WHO classification) located in the anterior cervical region (thyroid topography) that was submitted to ultrasound-guided fine-needle aspiration biopsy. The cytologic smears were moderately cellular to very cellular and were composed of numerous spindle-shaped cells placed predominantly in tissue fragments, with no specific arrangement. Isolated spindle-shaped cells were also present in a hemorrhagic background. The nuclei were elongated with finely granular chromatin and inconspicuous nucleoli. Mitosis, nuclear atypia, and necrosis were not detected. There was a scarce lymphoid population of small lymphocytes. The cytologic aspects, the correlation with histologic findings, and the differential cytologic diagnosis are reviewed. PMID- 15880712 TI - Fine-needle aspiration of ganglioneuroma. AB - Ganglioneuroma is a rare soft tissue neoplasm, arising most frequently in the mediastinum or retroperitoneum in children and young adults. Correct evaluation of this lesion is of crucial importance, as there are some malignant neoplasms that have similar clinical presentations. As reports of the cytologic appearance of ganglioneuroma are few, the author describes the cytologic features of three patients examined initially by fine-needle aspiration (FNA) with correlation to histology and clinical data. This paper shows that the cytologic features of ganglioneuromas can be diagnostic if both spindle cells and ganglion cells are present in smears. The absence of immature cells and necrosis/inflammation differentiates ganglioneuromas from their malignant counterparts, ganglioneuroblastomas and neuroblastomas. Immunocytochemistry can help to establish the neurogenic origin of the tumor cells. PMID- 15880713 TI - Fine-needle aspiration of a primary mediastinal large B-cell lymphoma: a case report with cytologic, histologic, and flow cytometric considerations. AB - Fine-needle aspiration (FNA) cytology and immunophenotyping by flow cytometry (FCM) are increasingly being used for diagnosing and subclassifying lymphoma in the REAL/WHO classification. Herein, we report a case of primary mediastinal large B-cell lymphoma (PMBL), a subtype of diffuse large B-cell lymphoma in the WHO classification, diagnosed by FNA cytology in conjunction with FCM. This, to our knowledge, has not previously been reported. A 57-yr-old woman presented with bilateral axillary lymphadenopathy and intermittent shortness of breath. CT scan revealed a 5-cm anterior mediastinal mass and mediastinal lymphadenopathy. Endoscopic ultrasound-guided FNA of a 4.5-cm subcarinal lymph node showed medium to large atypical lymphocytes with scant to moderate finely vacuolated cytoplasm. Nuclei were enlarged, cleaved, noncleaved, lobulated, and hyperchromatic. The background showed lymphoglandular bodies. Malignant large cell lymphoma was cytologically diagnosed. FCM, performed on a portion of the FNA specimen, demonstrated large B cells devoid of surface immunoglobulin expression, the characteristic immunophenotype of PMBL. The histologic diagnosis was PMBL. Touch imprint cytology of the histologic specimen showed large cells with a narrow rim of clear cytoplasm and prominent outer cell border. Nuclear features were similar to the FNA specimen. In the presence of a mediastinal mass, FNA cytology in conjunction with FCM can effectively diagnose PMBL in the appropriate clinical setting. PMID- 15880714 TI - Metastatic melanoma of the thyroid mimicking a papillary carcinoma in fine-needle aspiration. PMID- 15880715 TI - Pediatric renal-cell carcinoma. PMID- 15880716 TI - Desmoplastic round cell tumor of childhood: can cytology with immunocytochemistry serve as an alternative for tissue diagnosis? AB - There are limited reports on the cytology of desmoplastic small round cell tumors (DSRCT). Fine needle aspiration biopsy (FNAB) findings in seven aspirates from four cases of histologically and immunohistochemically confirmed cases were analyzed with the main intention of ascertaining if cytological diagnosis of DSRCT is possible. Also assessed were the immunocytochemistry(ICC) findings in these cases. The basic cytological impression was that of a cohesive small round cell tumor. Nuclei showed granular chromatin with grooves, nuclear molding and inconspicuous nucleoli. Stromal fragments were noted in all four cases. In two cases, awareness of cytological features in the appropriate clinical context led to a suggestion of the diagnosis of DSRCT on cytology itself. ICC on destained smears showed positivity for cytokeratin, epithelial membrane antigen (EMA), desmin and WT-1 in two cases. In conclusion, given the right clinical setting, a cytological diagnosis of DSRCT is plausible and in conjunction with ICC may help in documenting the polyphenotypic nature and thereby confirming the diagnosis. PMID- 15880717 TI - Clear-cell adenocarcinoma of the female genital tract: presence of hyaline stroma and tigroid background in various types of cytologic specimens. AB - Hyaline basement membrane-like stromal material and tigroid background are distinctive cytologic features observed in Diff-Quik (DQ)- or Giemsa-stained smears of clear-cell adenocarcinoma (CCA) of the female genital tract. However, it is uncertain how often these features are present in different types of cytologic specimens, and which type of preparation is optimal for this diagnosis. We therefore reviewed the cytologic features of CCA in three types of specimens, including 15 scrape cytology specimens, 7 fine-needle aspiration (FNA) specimens, and 15 peritoneal cytology specimens, with emphasis on the features observed in DQ-stained smears. The cell morphology in scrape cytology specimens and FNA specimens was comparable, whereas in peritoneal cytology specimens, the cytoplasm was better preserved. Most tumor cells had fragile cytoplasm containing variable amounts of fine vacuoles, and round nuclei with distinct or prominent nucleoli. Hyaline stroma was present in 93% of scrape cytology specimens, 71% of FNA specimens, and 80% of peritoneal cytology specimens. Tigroid background was observed in 47% of scrape cytology specimens, 43% of FNA specimens, but in none of the peritoneal cytology specimens. Formation of a tigroid background may be prevented by the abundant fluid content in peritoneal cytology specimens. Hyaline stroma and tigroid background were uncommonly seen in scrape smears from other types of primary ovarian tumors, mainly juvenile granulosa cell tumor and yolk sac tumor. However, the additional presence of papillary structures allows CCA to be readily distinguished from these other tumors. We propose that scrape cytology offers the best approach for the intraoperative cytologic diagnosis of CCA. PMID- 15880718 TI - Diagnostic intraoperative imprint cytology of a solid pseudopapillary tumor of the pancreas. PMID- 15880719 TI - Direct estimate of the conjugative and hyperconjugative stabilization in diynes, dienes, and related compounds. PMID- 15880720 TI - Radical mechanism of a place-exchange reaction of au nanoparticles. PMID- 15880721 TI - ATM mutations, haplotype analysis, and immunological status of Russian patients with ataxia telangiectasia. AB - Mutations in the ATM gene are responsible for the autosomal recessive disorder, ataxia telangiectasia (A-T). Mutations in different ethnic groups are distributed along the entire length of the large, 66 exon ATM gene. In this study, A-T patients from 16 Russian families were assessed for immunological status and ATM haplotype analysis, and screened for ATM mutations. Haplotype analysis was performed to enhance the efficiency of mutation detection. Mutations predicted to cause disease were identified in 19 of 32 alleles (59%), including a truncating mutation (c.5932G>T) that was identified in 8/32 (25%) alleles both by haplotype analysis and mutation screening. This mutation has been found in low abundance in other European A-T cohorts suggesting that this founder-effect mutation may be of Russian origin. The abundance of this mutation may allow for large-scale screening of cancer patients to help clarify the role of ATM in breast and other cancers. Nine of the remaining mutations were previously unreported, and add to the multitude of unique mutations found throughout the gene. PMID- 15880722 TI - Development of a resorbable macroporous cellulosic material used as hemostatic in an osseous environment. AB - The control of bleeding is a frequently encountered therapeutic problem, particularly during dental surgery. The most efficient substances used to resolve this problem are not risk-free because of their animal or human origins, so cellulosic materials are potentially of interest. The aim of this study was to develop a resorbable macroporous cellulosic material for use as a resorbable hemostatic agent in bone sites. The degradation and the cytocompatibility of the cellulosic material versus controls were evaluated and its behaviour in vivo was studied. An original process using calcium carbonate powder as inverse matrix was used to develop a macroporous material. In order to predegrade the cellulosic material for hemostatic use, oxidation was performed with periodate. A dialdehyde component unstable at physiological pH was thus obtained. The material was found to have cytotoxicity, biocompatibility, and resorption properties similar to control but its hemostatic power was higher. PMID- 15880723 TI - Novel and recurrent mutations in the C-terminal domain of COMP cluster in two distinct regions and result in a spectrum of phenotypes within the pseudoachondroplasia -- multiple epiphyseal dysplasia disease group. AB - Pseudoachondroplasia (PSACH) and some forms of multiple epiphyseal dysplasia (MED) result from mutations in the gene encoding cartilage oligomeric matrix protein (COMP). COMP is a large pentameric glycoprotein found predominantly in the extracellular matrix of cartilage, tendon, and ligament. As a modular protein, it is composed of a coiled-coil domain, four type II (T2) repeats, eight type III (T3) repeats, and a large globular C-terminal domain (CTD). The majority (>85%) of COMP mutations causing PSACH or MED are found in the exons encoding the T3 repeats, and the disease mechanism has been characterised in detail. Much less is known about disease-causing mutations in the CTD; in 10 years only seven mutations have been identified. In this study, we describe eight novel and two recurrent mutations that we have recently identified in patients with PSACH or MED. Interestingly, these mutations result in a spectrum of disease, ranging from mild MED to severe PSACH. Mapping of all known COMP CTD mutations on a three dimensional model of the C-terminal domain shows that the CTD mutations cluster in two distinct regions. These regions are probably important in stabilising the T3-CTD structure and mediating intra- or intermolecular interactions. PMID- 15880725 TI - Mechanism(s) of increased vascular cell adhesion on nanostructured poly(lactic-co glycolic acid) films. AB - Studies have shown that poly(lactic-co-glycolic acid) (PLGA) films with nanometer surface features promote vascular endothelial and smooth muscle cell adhesion. The objective of this in vitro research was to begin to understand the mechanisms behind this observed increase in vascular cell adhesion. Results provided evidence that nanostructured PLGA adsorbed significantly more vitronectin and fibronectin from serum compared to conventional (or those not possessing nanometer surface features) PLGA. When separately preadsorbing both vitronectin and fibronectin, increased vascular smooth muscle and endothelial cell density was observed on nanostructured (compared to conventional) PLGA. Additionally, blocking of cell-binding epitopes of fibronectin and vitronectin significantly decreased vascular cell adhesion on nanostructured (compared to conventional) PLGA. For this reason, results of the present in vitro study demonstrated that cell adhesive proteins adsorbed in different quantities and altered bioactivity on nanostructured compared to conventional PLGA topographies, which (at least in part) may account for the documented increased vascular cell adhesion on nanostructured PLGA. In this manner, this study continues to provide evidence for the promise of nanostructured PLGA in vascular tissue engineering applications. PMID- 15880726 TI - Neural adhesion molecules L1 and CHL1 are survival factors for motoneurons. AB - Many neurotrophic factors with survival activity for motoneurons in vivo were first identified using cultures of purified embryonic motoneurons. The L1 neural cell adhesion molecule has multiple roles in brain development. We showed by in situ hybridization and RT-PCR that L1 mRNA was expressed at significant levels in motoneurons of embryonic and postnatal spinal cord. We therefore cultured purified motoneurons from E14 rat embryos in the absence of trophic factors but with L1-Fc and CHL1-Fc fusion proteins. L1-Fc prevented the death of approximately half of the motoneurons that were saved by BDNF in a dose-dependent manner (EC50 = 10 pM). CHL1-Fc saved the same number of motoneurons as did L1-Fc, whereas P0-Fc had little neurotrophic activity at the same concentrations. Survival induced by L1 and CHL1 was completely inhibited by 20 microM LY294002 and PD98059, indicating that both MEK and PI3K pathways are required for signaling by these molecules. L1 can signal in other cell types through the FGF receptor FGFR1. In cultures of motoneurons, effects of suboptimal concentrations of L1 and suboptimal concentrations of FGF-2 were additive, but the effects of optimal concentrations of FGF-2 (50 ng/ml) were not further increased in the presence of L1-Fc. Thus, in this system, too, FGF and L1 may use similar signaling pathways. PMID- 15880727 TI - The spectrum of aldolase B (ALDOB) mutations and the prevalence of hereditary fructose intolerance in Central Europe. AB - We investigated the molecular basis of hereditary fructose intolerance (HFI) in 80 patients from 72 families by means of a PCR-based mutation screening strategy, consisting of heteroduplex analysis, restriction enzyme digest, DNA single strand electrophoresis, and direct sequencing. For a subset of patients mutation screening with DHPLC was established which turned out to be as fast and as sensitive as the more conventional methods. Fifteen different mutations of the aldolase B (ALDOB) gene were identified in HFI patients. As in smaller previous studies, p.A150P (65%), p.A175D (11%) and p.N335K (8%) were the most common mutated alleles, followed by c.360_363delCAAA, p.R60X, p.Y204X, and c.865delC. Eight novel mutations were identified in eight families with HFI: a small indel mutation (c.1044_1049delTTCTGGinsACACT), two small deletions (c.345_372del28; c.841_842delAC), two splice site mutations (c.113-1G>A, c.799+2T>A), one nonsense mutation (c.612T>G (p.Y204X)), and two missense mutations (c.532T>C (p.C178R), c.851T>C (p.L284P)). By mutation screening for the three most common ALDOB mutations by DHPLC in 2,000 randomly selected newborns we detected 21 heterozygotes. Based on these data and after correction for less common and private ALDOB mutations, HFI prevalence in central Europe is estimated to be 1:26,100 (95% confidence interval 1: 12,600-79,000). PMID- 15880728 TI - Metastases to the kidney: eleven cases diagnosed by aspiration biopsy with histological correlation. AB - Metastases to the kidney from extrarenal primary tumors are uncommon and may mimic renal-cell carcinoma clinically when presenting as a single mass with hematuria. Fine-needle aspiration biopsy (FNAB) is a useful diagnostic method for the evaluation of primary renal tumors. Only a few studies have investigated the value of cytological evaluation of secondary renal tumors. We report our experience with these tumors. Eleven cases of extrarenal primary tumors metastatic to the kidney, diagnosed by aspiration biopsy with histological correlation, are discussed. The diagnosis of metastatic disease to the kidney was accurately made by aspiration biopsy. Knowledge of the patients' history, histological correlation with the primary tumor, and the radiological characteristics of the renal masses were helpful in achieving a correct diagnosis. FNA cytology (FNAC) is an accurate method for the diagnosis of tumors metastatic to the kidney. Distinction between primary and secondary tumors of the kidney is crucial to guide management and prevent unnecessary surgery. PMID- 15880729 TI - Side-by-side patterning of multiple alkanethiolate monolayers on gold by edge spreading lithography. PMID- 15880730 TI - Synthesis of ultrathin zinc nanowires and nanotubes by vapor transport. PMID- 15880731 TI - Application of genomewide SNP arrays for detection of simulated susceptibility loci. AB - The prospect of SNP-based genomewide association analysis has been extensively discussed, but practical experiences remain limited. We performed an association study using a recently developed array of 11,555 SNPs distributed throughout the human genome. A total of 104 DNA samples were hybridized to these chips with an average call rate of 97% (range 85.3-98.6%). The resulting genomewide scans were applied to distinguish between carriers and noncarriers of 37 test variants, used as surrogates for monogenic disease traits. The test variants were not contained in the chip and had been determined by other methods. Without adjustment for multiple testing, the procedure detected 24% of the test variants, but the positive predictive value was low (2%). Adjustment for multiple testing eliminated most false-positive associations, but the share of true positive associations decreased to 10-12%. We also simulated fine-mapping of susceptibility loci by restricting testing to the immediate neighborhood of test variants (+/-5 Mb). This increased the proportion of correctly identified test variants to 22-27%. Simulation of a bigenic inheritance reduced the sensitivity to 1%. Similarly adverse effect had reduction of allelic penetrance. In summary, we demonstrate the feasibility and considerable specificity of SNP array-based association studies to detect variants underlying monogenic, highly penetrant traits. The outcome is affected by allelic frequencies of chip SNPs, by the ratio between simulated "cases" and "controls," and by the degree of linkage disequilibrium. A major improvement is expected from raising the density of the SNP array. PMID- 15880732 TI - DNA damage in leukocytes of workers occupationally exposed to arsenic in copper smelters. AB - Inorganic arsenic (i-As) is a known human carcinogen; however, humans continue to be exposed to i-As in drinking water and in certain occupational settings. In this study, we used the Comet assay to evaluate DNA damage in the somatic cells of workers from three Polish copper smelters who were occupationally exposed to i As. Blood samples were collected from 72 male workers and 83 unexposed male controls and used for the detection of DNA damage, oxidative DNA damage, and DNA damage after a 3-hr incubation in culture. Urine samples were collected to assess the level of exposure. The mean concentration of arsenic metabolites in urine [the sum of arsenite (AsIII), arsenate (AsV), monomethylarsenate (MMA) and dimethylarsenate (DMA)] and the concentrations of DMA (the main metabolite in urine) were higher in workers than in controls, but the differences were not statistically significant. By contrast, the level of DNA damage, expressed as the median tail moment, was significantly higher in the leukocytes of workers than in the controls. Comet assays conducted with formamidopyrimidine glycosylase (FPG) digestion to detect oxidative DNA damage indicated that oxidative lesions were present in leukocytes from both the exposed and control groups, but the levels of damage were significantly higher among the workers. Incubation of the cells in culture resulted in a significant reduction in the levels of DNA damage, especially among leukocytes from the workers, suggesting that the DNA damage was subject to repair. Our findings indicate that copper smelter workers have increased levels of DNA damage in somatic cells, suggesting a potential health risk for the workers. Although i-As was present in air samples from the smelters and in urine samples from workers, no clear association could be made between i As exposure and the DNA damage. PMID- 15880733 TI - Genotoxicity of drinking water disinfectants in plant bioassays. AB - The genotoxicity of two widely used drinking water disinfectants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO(2)), and a new disinfectant, peracetic acid (PAA, CH(3)-CO-COOH), was evaluated in three short-term plant tests: (1) induction of anaphase chromosome aberrations in the root cells of Allium cepa, (2) micronucleus induction in the root cells of Vicia faba, and (3) micronucleus induction in Tradescantia pollen cells. The study was carried out in the laboratory by directly exposing the plants to several concentrations of the disinfectants in redistilled water at unadjusted (acid) and adjusted (neutral) pHs. Both 0.1 and 0.2 mg/l NaClO induced chromosome aberrations in the Allium cepa test at acid pH, but concentrations up to 0.5 mg/l of all the disinfectants were negative at neutral pH. Concentrations ranging from 0.1 to 0.5 mg/l NaClO, ClO(2,) and PAA induced micronuclei in Vicia faba at acid pH, while 1-2 mg/l NaClO and ClO(2) and 0.5-2 mg/l PAA gave positive responses at neutral pH. Most of concentrations of ClO(2) produced positive responses in the Tradescantia micronucleus test. In general, the highest levels of genotoxicity were observed under acid conditions; at acid pH, significant effects were induced by low concentrations of ClO(2) and PAA. Since the test concentrations of disinfectants are typical of those encountered in the biocidal treatment of tap water and similar concentrations are consumed daily by a large number of people, the genotoxicity of these compounds may constitute a significant public health concern. PMID- 15880734 TI - Induction of cell proliferation, micronuclei and hyperdiploidy/polyploidy in the mammary cells of DDT- and DMBA-treated pubertal rats. AB - The environmental estrogen, dichlorodiphenyltrichloroethane (DDT), and its metabolites have been implicated in the development of breast cancer through mechanisms that remain to be elucidated. It has been hypothesized that exposure to DDT and its metabolites, during critical periods of development, can contribute to an elevated risk for breast cancer in adults. In the present study, we have investigated the effect of o,p'-DDT on mammary gland cell proliferation and chromosomal alterations, in a rat mammary cancer model (commonly used to study human cancer), to gain insights into its potential role in the development of breast cancer. Twenty-one-day-old female Sprague-Dawley (SD) rats were administered o,p'-DDT, 7,12-dimethylbenz[a]anthracene (DMBA), genistein, DDT+DMBA, or DDT+DMBA+genistein, over a 14-day period. To determine changes in chromosome number and structure, we used the micronucleus assay as well as multicolor fluorescence in situ hybridization (FISH) region-specific DNA probes for rat chromosomes 4 and 19. Cell proliferation was evaluated using 5-bromo-2' deoxyuridine (BrdU). Significant increases in BrdU-incorporated cells were seen in the rats treated with DDT+DMBA. Although micronucleus frequencies were somewhat elevated in several of the treatment groups, significant increases were not seen in any of them. Significant increases in numerical chromosomal aberrations were detected in all of the DDT- and DMBA-treated groups. Genistein significantly reduced BrdU incorporation and polyploidy in the DDT+DMBA-treated rats. These initial studies indicate that DDT and DMBA can induce cellular and chromosomal alterations in the rat mammary gland, which is consistent with the hypothesis that these agents can induce early events in mammary carcinogenesis. PMID- 15880735 TI - Further development of the beta-lactamase MutaGen assay and evaluation by comparison with Ames fluctuation tests and the umu test. AB - A rapid, high-throughput bacterial mutagenicity test system has been developed (MutaGen test) that detects reversions of inactivating base-pair substitutions and frameshifts in a TEM-1 class A beta-lactamase (ampicillinase) gene. To quickly and sensitively detect mutagens, the system utilises a series of plasmids that contain the mutated ampicillinase gene and the mucAB operon. Inactivating mutations in the ampicillinase gene include frameshifts integrated into repetitive GC-sequences and G-runs known to be mutagenic hot-spots, and base-pair substitutions inserted in or around the beta-lactamase active site. Frameshift mutations completely inactivated the enzyme only when located downstream of the active-site serine (Ser68). Previous (reporter gene based) assays with this system have detected reversion to ampicillin resistance by luminescence driven by induction of the tet-promotor controlled lacZ gene. In the present study, we describe the construction and evaluation of 19 additional potential tester strains. We also developed conditions for detecting reversions by pH shift using bromocresol purple and by directly detecting the enzymatic activity of beta lactamase using nitrocefin. A 384-well format version of the pH shift MutaGen test was used to assay more than 20 chemicals. The responses in the assay were compared with responses for the same chemicals in the umu test and Ames fluctuation assays. The results indicate that the MutaGen test has high specificity for detecting specific mutations and, in some instances, better sensitivity than the other tests. Since the test is easy to conduct, sterile working conditions are not necessary, and the mutagenicity results are available either within one working day or overnight, the assay shows promise for the rapid screening of potentially genotoxic substances. PMID- 15880736 TI - Antimutagenicity of green tea polyphenols in the liver of transgenic medaka. AB - We examined the ability of a mixture of the predominant green tea polyphenolic compounds (GTP) to reduce benzo[a]pyrene (B[a]P)-induced mutations in the cII gene of the lambda transgenic medaka. Fish were treated with 50 ppb B[a]P for 24 hr, followed by exposure to 2 ppm or 10 ppm GTP for 28 days. cII mutations in livers of fish exposed to B[a]P were increased significantly, 2.6-fold above controls. In contrast, the addition of GTP significantly reduced the frequency of cII mutants by 84%, comparable to that of controls. The frequencies of mutations at G:C basepairs, mutations that are highly characteristic of B[a]P exposure, were elevated significantly in treated fish. By comparison, B[a]P-exposed fish also treated with GTP showed reductions in these mutations, demonstrating a protective effect of GTP against B[a]P-induced mutagenesis. The antioxidant mechanism of GTP possibly played an important role in the reduction of B[a]P mutagenicity. These results corroborate findings from rodent models, showing that the protective effects of green tea extend to different species, and suggesting that similar mechanisms of B[a]P mutagenesis and GTP antimutagenesis are shared among the models. These studies illustrate the utility of lambda transgenic medaka for in vivo mutation analyses and suggest that this fish may be a valuable model in chemoprevention studies. PMID- 15880737 TI - Genotoxicity and physicochemical characteristics of traffic-related ambient particulate matter. AB - Exposure to ambient particulate matter (PM) has been linked to several adverse health effects. Since vehicular traffic is a PM source of growing importance, we sampled total suspended particulate (TSP), PM(10), and PM(2.5) at six urban locations with pronounced differences in traffic intensity. The mutagenicity, DNA adduct formation, and induction of oxidative DNA damage by the samples were studied as genotoxicological parameters, in relation to polycyclic aromatic hydrocarbon (PAH) levels, elemental composition, and radical-generating capacity (RGC) as chemical characteristics. We found pronounced differences in the genotoxicity and chemical characteristics of PM from the various locations, although we could not establish a correlation between traffic intensity and any of these characteristics for any of the PM size fractions. Therefore, the differences between locations may be due to local sources of PM, other than traffic. The concentration of total (carcinogenic) PAHs correlated positively with RGC, direct and S9-mediated mutagenicity, as well as the induction of DNA adducts and oxidative DNA damage. The interaction between total PAHs and transition metals correlated positively with DNA-adduct formation, particularly from the PM(2.5) fraction. RGC was not associated with one specific PM size fraction, but mutagenicity and DNA reactivity after metabolic activation were relatively high in PM(10) and PM(2.5), when compared with TSP. We conclude that the toxicological characteristics of urban PM samples show pronounced differences, even when PM concentrations at the sample sites are comparable. This implies that emission reduction strategies that take chemical and toxicological characteristics of PM into account may be useful for reducing the health risks associated with PM exposure. PMID- 15880738 TI - A synthetic block copolymer regulates S1 nuclease fragmentation of supercoiled plasmid DNA. PMID- 15880739 TI - A single-crystal-to-single-crystal transformation mediated by argentophilic forces converts a finite metal complex into an infinite coordination network. PMID- 15880740 TI - Morphology and evolution of the jaw suspension in lamniform sharks. AB - The morphology of the jaw suspension and jaw protrusion mechanism in lamniform sharks is described and mapped onto a cladogram to investigate how changes in jaw suspension and protrusion have evolved. This has revealed that several evolutionary modifications in the musculoskeletal apparatus of the jaws have taken place among lamniform sharks. Galeomorph sharks (Carcharhiniformes, Lamniformes, Orectolobiformes, and Heterodontiformes) have paired ethmopalatine ligaments connecting the ethmoid process of the upper jaw to the ethmoid region of the cranium. Basal lamniform sharks also acquired a novel single palatonasal ligament connecting the symphysis of the upper jaw to the cranium mid-ventral to the nasal capsule. Sharks in the family Lamnidae subsequently lost the original paired ethmopalatine ligament while retaining the novel palatonasal ligament. Thus, basal lamniform taxa (Mitsukurina owstoni, Carcharius taurus, Alopias vulpinnis) have increased ligamentous support of the lateral region of the upper jaw while derived species (Lamnidae) have lost this lateral support but gained anterior support. In previous studies the morphology of the jaw suspension has been shown to play a major role in the mechanism of upper jaw protrusion in elasmobranchs. The preorbitalis is the primary muscle effecting upper jaw protrusion in squalean (sister group to galeomorphs) and carcharhiniform (sister group to lamniforms) sharks. The preorbitalis originates from the quadratomandibularis muscle and inserts onto the nasal capsule in squalean and carcharhiniform sharks. Carcharhiniform sharks have evolved a subdivided preorbitalis muscle with the new division inserting near the ethmoid process of the palatoquadrate (upper jaw). Alopid sharks have also independently evolved a partially subdivided preorbitalis with the new division inserting at the base of the ethmoid process and surrounding connective tissue. Lamnid sharks have retained the two preorbitalis divisions but have modified both of the insertion points. The original ventral preorbitalis division now inserts onto the connective tissue surrounding the mid-region of the upper jaw, while the new dorsal preorbitalis division inserts onto the surrounding connective tissue and skin at a more posterior position on the upper jaw. The retractor muscle of the jaws, the levator hyomandibularis, has also been modified during the evolution of lamniform sharks. In most sharks, including basal lamniforms, the levator hyomandibularis inserts onto the hyomandibula and functions to retract the jaws after protrusion. In alopid and lamnid sharks the levator hyomandibularis inserts primarily onto the upper and lower jaws around the jaw joint and is a more direct route for retracting the jaws. Thus, there has been at least one instance of character loss (ethmopalatine ligament), acquisition (palatonasal ligament), subdivision (preorbitalis), and modification (ventral preorbitalis, dorsal preorbitalis, and levator hyomandibularis) in the ligaments and muscles associated with the jaw suspension and jaw protrusion mechanism in lamniform sharks. While derived lamniform sharks (Lamna nasus, Carcharodon carcharius, and Isurus oxyrinchus) lost the ancestral passive lateral support of the ethmoid articulation of the upper jaw, they simultaneously acquired muscular support by way of the levator hyomandibularis, which provides a dynamic mechanism for lateral support. The evolution of multiple divisions of preorbitalis insertions onto the palatoquadrate and modification of the levator hyomandibularis insertion directly onto the jaws provides an active mechanism for multiple protractions and retractions of the upper jaw, which is advantageous in those sharks that gouge or saw pieces from large oversized prey items. PMID- 15880741 TI - Amplification and overexpression of aurora kinase A (AURKA) in immortalized human ovarian epithelial (HOSE) cells. AB - Immortalization is an early and essential step of human carcinogenesis. Amplification of chromosome 20q has been shown to be a common event in immortalized cells and cancers. We have previously reported that gain and amplification of chromosome 20q is a non-random and common event in immortalized human ovarian surface epithelial (HOSE) cells. The chromosome 20q harbors genes including TGIF2 (20q11.2-q12), AIB1 (20q12), PTPN1 (20q13.1), ZNF217 (20q13.2), and AURKA (20q13.2-q13.3), which were previously reported to be amplified and overexpressed in ovarian cancers. Some of these genes may be involved in immortalization of HOSE cells and represent crucial premalignant changes in ovarian surface epithelium. Investigation of the involvement of these genes was examined in four pairs of pre-crisis (preimmortalized) and post-crisis (immortalized) HOSE cells. Overexpression of AURKA (Aurora kinase A), also known as BTAK and STK15, by both real time-quantitative polymerase chain reaction (RT QPCR) and Western blotting was detected in all the four immortalized HOSE cells examined while overexpression of AIB1 and ZNF217 was observed in two of four immortalized HOSE cells examined. Overexpression of TGIF2 and PTPN1 was not significant in our immortalized HOSE cell systems. The degree of overexpression of AURKA was shown to be closely associated with the amplification of chromosome 20q in immortalized HOSE cells. Fluorescence in situ hybridization (FISH) with labeled P1 artificial clone (PAC) confirmed the amplification of the chromosomal region (20q13.2-13.3) where AURKA resides. DNA amplification of AURKA was also confirmed using semi-quantitative PCR. Our study showed that amplification and overexpression of AURKA is a common and significant event during immortalization of HOSE cells and may represent an important premalignant change in ovarian carcinogenesis. PMID- 15880742 TI - Chronic treatment with the mGlu5R antagonist MPEP reduces the functional effects of the mGlu5R agonist CHPG in the striatum of 6-hydroxydopamine-lesioned rats: possible relevance to the effects of mGlu5R blockade in Parkinson's disease. AB - This study was designed to test whether chronic treatment with the metabotropic glutamate receptor 5 (mGlu5R) antagonist MPEP showed antiparkinsonian effects in rats unilaterally lesioned with 6-hydroxydopamine (6-OHDA) (a "classic" model of Parkinson's disease, PD), and to evaluate whether chronic MPEP influenced the functional properties and/or the expression of striatal mGlu5Rs. Wistar rats were lesioned with 6-OHDA and then treated with MPEP (3 mg/kg/day, i.p.) or its vehicle over 2 weeks. Chronic MPEP did not induce measurable antiparkinsonian effects, since no differences were found between MPEP- and vehicle-treated animals in the pattern of L-DOPA-induced contralateral rotations. In corticostriatal slices taken from animals chronically treated with MPEP, the functional effects of the mGlu5R agonist CHPG were significantly reduced in the lesioned vs. the intact side, while no changes were found in slices taken from vehicle-treated rats. The binding of [3H]MPEP to striatal membranes showed that neither the maximal number of binding sites (Bmax) nor the dissociation constant (Kd) were changed by the lesion and/or by chronic MPEP. While chronic MPEP did not potentiate L-DOPA-induced turning in a classical model of PD, its ability to reduce mGlu5R-associated signal could help to explain the neuroprotective/antiparkinsonian effects observed in other models of PD. PMID- 15880743 TI - Neuronal vulnerability in mouse models of Huntington's disease: membrane channel protein changes. AB - Huntington's disease (HD) is caused by a polyglutamine expansion that results in atrophy of the striatum and frontal cortex during disease progression. HD susceptible striatal neurons are affected chronologically with initial degeneration of the striatopallidal neurons then the striatonigral projections, whereas large aspiny striatal interneurons (LAN) survive. Two classes of critical membrane proteins were evaluated in transgenic mouse models to determine their association with HD susceptibility, which leads to dysfunction and death in selected striatal neuron populations. We examined potassium (K+) channel protein subunits that form membrane ionophores conducting inwardly and outwardly rectifying K+ currents. K+ channel protein staining was diminished substantially in the HD striatal projection neurons but was not expressed in the HD-resistant LAN. Because loss of K+ channel subunits depolarizes neurons, other voltage-gated ionophores will be affected. N-methyl-D-aspartate (NMDA) receptors and their phosphorylation by cyclic AMP were studied as a mechanism contributing to excitotoxic vulnerability in striatal projection neurons that would lose voltage regulation after diminished K+ channels. NR1 subunits showed significant elevation in the HD transgenic projection systems but were expressed at very low levels in LAN. NR1 subunit phosphorylation by cyclic AMP also was enhanced in striatal projection neurons but not in LAN. Cyclic AMP-driven phosphorylation of NMDA receptors increases the channel open time and elevates neuronal glutamate responsiveness, which may lead to excitotoxicity. Together our data suggest that changes in these proteins and their modification may predispose striatal projection neurons to dysfunction and then degeneratation in HD and provide a mechanism for LAN resistance in the disease. PMID- 15880744 TI - Colorectal carcinoma screening among Hispanics and non-Hispanic whites in a rural setting. AB - BACKGROUND: Colorectal carcinoma ranks as the second most common cancer and the second leading cause of cancer death in the United States. Hispanics are less likely than their non-Hispanic white counterparts to have ever received a fecal occult blood test (FOBT) or sigmoidoscopy/colonoscopy. Little is known about the barriers to screening in the Hispanic population. METHODS: The authors used baseline data from a community randomized trial of cancer prevention to compare screening prevalence and the associations between reported barriers and screening participation between Hispanics (n = 137) and non-Hispanic whites (n = 491) age > or = 50 years. RESULTS: Hispanics were less likely than non-Hispanic whites to have ever received an FOBT (P = 0.003) or sigmoidoscopy/colonoscopy (P = 0.001). No significant difference across ethnic groups was observed in the prevalence of recent screening using FOBT (29.8% for Hispanics vs. 34.5% for non-Hispanic whites; P = 0.41), but recent use of sigmoidoscopy/colonoscopy was lower for Hispanics (24.1% for Hispanics vs. 33.7% for non-Hispanic whites; P 0.06). Lacking health care coverage or having few years of education were directly associated with failure to ever receive an FOBT or sigmoidoscopy/colonoscopy. CONCLUSIONS: Interventions to improve adherence to colorectal carcinoma screening recommendations among Hispanics should target initial screening examinations, particularly among those lacking health care coverage or having low levels of education. PMID- 15880745 TI - Studies of reduced folate carrier 1 (RFC1) A80G and 5,10 methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms with neural tube and orofacial cleft defects. PMID- 15880746 TI - On ubiquitin ligases and cancer. AB - Protein kinase genes account for almost 10% of all currently known cancer genes, highlighting the role of signal transduction in oncogenesis. A reexamination of the literature and available databases shows that E3 ubiquitin ligases are also key mediators of tumorigenesis. Altogether kinase and E3 genes represent more than 15% of the known cancer genes, underlining the importance of phosphorylation and ubiquitylation signaling pathways in cancer formation. Considering the recent literature reporting correlations between alterations in ubiquitylation processes and oncogenesis, this percentage is likely to increase even further in the future. Finally, E3 genes could serve as baits for the identification of additional cancer genes (e.g. their interacting partners). In contrast, deubiquitinases, like phosphatases, are not overrepresented among cancer genes. The same holds for E1 and E2 genes. Thus, kinase and E3 genes represent primary targets as cancer susceptibility genes for mutation screening and for the design of novel therapies. PMID- 15880747 TI - Functional analysis of natural mutations in two TWIST protein motifs. AB - The basic helix-loop-helix protein Twist, a transcriptional repressor, is essential for embryogenesis in both invertebrates and vertebrates. Haploinsufficiency of the human TWIST1 gene, which causes the craniosynostosis disorder Saethre-Chotzen syndrome (SCS), is related to failure to repress transcription of CDKN1A (which encodes p21/WAF1/CIP1), promoting osteoblast differentiation. We have examined the functional significance of natural TWIST1 variants present in craniosynostosis patients and in their healthy relatives. Both deletion and duplication variants of the glycine-rich tract Gly5AlaGly5 inhibited E2A (E12/E47)-dependent transcription of CDKN1A to a similar degree as wild-type protein, indicating that the length of this glycine tract is not critical for efficient transcriptional repression. We also evaluated a newly identified heterozygous TWIST1 variant (c.115C>G, encoding p.Arg39Gly), located within a putative nuclear localization signal (NLS), that was present in a child with mild SCS and her clinically unaffected father and grandmother. Unlike wild type protein, this mutant required cotransfected E12 to localize to the nucleus, indicating that the NLS, including amino acid 39, is essential for nuclear localization; inhibition of E2A-dependent transcription of CDKN1A occurred normally. This analysis further dissects the structure-function relationships of TWIST and corroborates with phenotypic observations of disease expressivity. PMID- 15880748 TI - The role of precursor-decomposition kinetics in silicon-nanowire synthesis in organic solvents. PMID- 15880749 TI - DNA-encoded self-assembly of gold nanoparticles into one-dimensional arrays. PMID- 15880750 TI - Radiotherapy alone or combined with surgery for salivary gland carcinoma. AB - BACKGROUND: The authors analyzed the outcomes of patients treated with radiotherapy (RT) alone or combined with surgery for carcinoma of the salivary glands. METHODS: Between October 1964 and June 2003, 224 previously untreated patients were treated with curative intent with RT alone (n = 64) or combined with surgery (n = 160) at the University of Florida College of Medicine (Gainesville, FL). The median follow-up period was 5.0 years (range, 0.4-31.6 years). RESULTS: The 10-year local control rate was 75%. Multivariate analysis of local control revealed that T classification (P < 0.0001) and treatment group (P < 0.0001) impacted this end point. Patients treated with surgery and adjuvant RT had improved local control compared with patients treated with RT alone. The 10 year locoregional control rate was 68%. Multivariate analysis of locoregional control revealed that overall stage (P < 0.0001) and treatment group (P = 0.0002) significantly influenced this end point. The 10-year distant metastasis-free survival rate was 68%. Multivariate analysis of distant metastasis-free survival revealed that overall stage (P < 0.0001) significantly influenced this end point. The 10-year cause-specific and overall survival rates were 57% and 44%, respectively. Multivariate analysis of cause-specific survival revealed that overall stage (P < 0.0001) significantly impacted this end point. Twenty-three patients (10%) experienced severe complications. CONCLUSIONS: A substantial proportion of patients with salivary gland carcinoma were cured with surgery combined with adjuvant RT. RT alone was used for patients with unresectable tumors and cured approximately 20% of those with advanced-stage disease. The probability of cure was influenced by the extent of disease and treatment group. PMID- 15880751 TI - Nanoscaffold mediates hydrogen release and the reactivity of ammonia borane. PMID- 15880752 TI - Copper-mediated selective oxidation of a C-H bond. PMID- 15880753 TI - Sequence of abnormal dendritic spine development in primary somatosensory cortex of a mouse model of the fragile X mental retardation syndrome. AB - Anatomical analyses of occipital and temporal cortex of patients with fragile X mental retardation syndrome (FXS) and in a mouse model of the syndrome (FraX mice) compared to controls have suggested that the fragile X mental retardation protein (FMRP) is important for normal spine structural maturation and pruning. However, a recent analysis of spine properties in somatosensory cortex of young FraX mice has suggested that this region may not exhibit spine abnormalities. While spine abnormalities were present 1 week after birth in somatosensory cortex, by 4 weeks almost all spine abnormalities had disappeared, suggesting that adult spine abnormalities observed in other cortical regions may not persist post-developmentally in somatosensory cortex. To resolve this discrepancy we examined spine properties in somatosensory cortex of young (day 25) and adult (day 73-76) FraX compared to wild-type (WT) mice. Spine properties in young FraX and WT mice did not consistently differ from each other, consistent with the recent analysis of developing somatosensory cortex. However, adult FraX mice exhibited increased spine density, longer spines, more spines with an immature appearing structure, fewer shorter spines, and fewer spines with a mature structure, a pattern consistent with prior analyses from other adult cortical brain regions in humans and mice. These findings (1) support the previous report of the absence of major spine abnormalities in the fourth postnatal week, (2) demonstrate normal spine development in WT mice, (3) demonstrate abnormal spine development after the fourth postnatal week in FraX mice, and (4) demonstrate spine abnormalities in somatosensory cortex of adult FraX compared to adult WT mice. In doing so, these findings resolve a potential conflict in the literature and more thoroughly describe the role of FMRP in spine development. PMID- 15880754 TI - High-level expression of fatty acid synthase in human prostate cancer tissues is linked to activation and nuclear localization of Akt/PKB. AB - Many human epithelial cancers, particularly those with a poor prognosis, express high levels of fatty acid synthase (FAS), a key metabolic enzyme linked to the synthesis of membrane phospholipids in cancer cells. In view of the recent finding that in the human prostate cancer cell line LNCaP, overexpression of FAS can be largely attributed to constitutive activation of the phosphatidylinositol 3 (PI3) kinase/Akt kinase pathway, the activation status of the Akt pathway, and whether this activation coincides with increased FAS expression, was examined in clinical prostate cancer tissues. Using well-preserved frozen prostatic needle biopsies and a sensitive Envision detection technique, S473-phosphorylated Akt (pAkt) was found in 11/23 low-grade prostatic intraepithelial neoplasia (PIN) lesions, in all (36/36) high-grade PINs, and in all (86/86) invasive carcinomas. Non-neoplastic tissues were negative. Interestingly, in low-grade PINs and low grade carcinomas, pAkt was mainly cytoplasmic or membrane-bound and was associated with moderate elevation of FAS expression. In 24/36 high-grade PINs and 82/88 invasive carcinomas, pAkt was found at least partly in the nucleus. Greater nuclear pAkt staining, and higher FAS expression, correlated with a higher Gleason score. In the light of previous findings that pAkt plays a causative role in the overexpression of FAS in cancer cells in culture, these data strongly suggest that high-level expression of FAS in prostate cancer tissues is linked to phosphorylation and nuclear accumulation of Akt. PMID- 15880755 TI - Photo-cross-linked small-molecule affinity matrix for facilitating forward and reverse chemical genetics. PMID- 15880756 TI - Membraneous localization of Jpk is not essential to exert cytotoxicity in F9 teratocarcinoma cells. AB - A novel gene, Jopock (Jpk), which was isolated as a trans-acting factor associating with the PSRE of murine Hoxa-7, has been shown to be toxic to both prokaryotic and eukaryotic cells when overexpressed. Here we demonstrate that the overexpression of enhanced green fluorescent protein (EGFP)-tagged Jpk in F9 cells results in the induction of apoptosis, as indicated by phosphatidylserine exposure, DNA fragmentation, and the alteration of mitochondria transmembrane potential. Fluorescence microscopy showed that EGFP-fused Jpk was mainly localized in the endoplasmic reticulum (ER) and a small amount was found in the mitochondria. Deletion mutants with a transmembrane (TM) domain showed a distribution similar to that of EGFP-Jpk, whereas constructs with a deletion of the TM domain localized in the whole cells. Deletion mapping experiments showed that Jpk with an N-terminal part deleted stimulated apoptosis to almost the same extent as that of the wild-type Jpk, indicating that the localization of Jpk in the ER and the TM domain does not appear to be essential for inducing cytotoxicity. Overall, these results suggest that Jpk, particularly the C terminal part of Jpk and/or 3'UTR, triggers apoptosis through a perturbation of mitochondrial membrane permeabilization. PMID- 15880757 TI - Trimeric G proteins in crustacean (Callinectes sapidus) Y-organs: occurrence and functional link to protein synthesis. AB - Crustacean Y-organs produce ecdysteroid molting hormones. Regulation of ecdysteroidogenesis appears to be complex, involving regulatory ligands (including but not limited to molt-inhibiting hormone, an eyestalk neurohormone) and the capacity of the Y-organs to respond to those ligands. Available data indicate cell signaling pathways involving cAMP, cGMP, or both may be involved in regulation of Y-organ function. Trimeric G proteins link receptor occupancy to regulation of intracellular cAMP levels. In studies reported here, we have assessed the occurrence of G proteins in blue crab (Callinectes sapidus) Y organs, and the link of G proteins to Y-organ function. Bacterial toxin-catalyzed ADP-ribosylation revealed a PTX-sensitive (alpha i-like) protein in Y-organ membranes, but failed to reveal a CTX-sensitive (alpha s-like) protein in Y-organ membranes. Western blotting with primary antibodies raised against conserved regions of mammalian G proteins detected an alpha i-immunoreactive protein (approximately 40 kDa) and two alpha s-immunoreactive proteins (approximately 50 and approximately 57 kDa) in Y-organ membrane preparations. Incubation of Y-organ membrane fractions with cholera toxin significantly suppressed incorporation of [35S]-methionine into TCA-precipitable Y-organ proteins, but had no detectable effect on ecdysteroidogenesis in short-term (6 h) incubations. The combined results indicate that C. sapidus Y-organs possess both Gi and Gs proteins, and that alpha s is functionally linked to regulation of glandular protein synthesis. PMID- 15880758 TI - Evidence that estrogen regulates the sex change of honeycomb grouper (Epinephelus merra), a protogynous hermaphrodite fish. AB - Circulating estradiol-17beta (E2) levels decrease precipitously during female to male (protogynous) sex change in fish. Whether this drop in E2 levels is a cause or consequence of sex change is still largely unknown. The present study treated adult female honeycomb groupers (Epinephelus merra) with aromatase inhibitor (AI, Fadrozole), either alone or in combination with E2, to investigate the role of estrogen in protogynous sex change. Control fish had ovaries undergoing active vitellogenesis; the gonads of AI-treated fish had already developed into testes, which produced sperm capable of fertilization. In contrast, co-treatment of fish with E2 completely blocked AI-induced sex reversal. AI treatment significantly reduced circulating levels of E2, whereas the addition of E2 to AI prevented the loss. The plasma androgen (testosterone and 11-ketotestosterone) levels were increased in the AI-treated fish, while the levels in the E2-supplemented fish were low compared to controls. Present results show that E2 plays an important role in maintaining female sex of hermaphrodite fishes, and that the inhibition of E2 synthesis causes oocyte degeneration leading to testicular differentiation in the ovary. PMID- 15880759 TI - The POCER indicator: a decision tool for non-agricultural pesticide use. AB - Pesticides are used not only in agriculture but also by public services and households. Much information is available on the influence of agricultural applications on human health by occupational exposure and on the environment. There is still a need for adequate information on the impact of non-agricultural applications of pesticides. The POCER indicator (Pesticide Occupational and Environmental Risk indicator) has been developed at Ghent University for agricultural situations, as a tool for applicators and decision-makers, by calculating the impact of pesticide treatments on the applicator, the worker, the bystander, groundwater, surface water, bees, earthworms, birds, useful arthropods and persistence in soil. A few adaptations in the POCER calculation method can make the indicator also very useful for non-agricultural conditions. The impact of plant protection products on human health and environment in public services and households can be calculated and the scores can be compared with each other, resulting in an improved pesticide programme. Decision makers might use the POCER indicator to evaluate the reduction programme of public services and communities and offer them measurements to reduce the impact of pesticides. For example, wearing protective clothing or taking into account the distance between application and surface water can greatly reduce the impact score. PMID- 15880760 TI - Cytotoxic T cells are preferentially activated in the duodenal epithelium from patients with florid coeliac disease. AB - Villous atrophy and increased numbers of intraepithelial T cells in duodenal biopsies represent a hallmark of coeliac disease. In the present study, an attempt has been made to define whether cytotoxic cell subsets are activated in situ in the affected mucosa of susceptible individuals early after ingestion of a gluten-containing diet. Duodenal biopsies from 11 patients with coeliac disease who repeatedly underwent endoscopic biopsy after ingestion of individually dosed amounts of gluten were used for immunohistochemistry and in situ hybridization. To identify the cell subsets expressing perforin mRNA and protein, in situ hybridization and FACS analyses were performed on cells isolated from fresh biopsies. Compared with normal mucosa, the number of intraepithelial lymphocytes containing perforin mRNA and protein increased significantly in tissue samples showing moderate or florid coeliac disease and closely paralleled the severity of morphological alteration, whereas the frequency of perforin-expressing lamina propria lymphocytes increased only moderately. Cells isolated from florid biopsies that expressed perforin mRNA and protein were preferentially T-cell receptor (TCR) alphabeta T cells. The increase in both the absolute number and the percentage of lymphocytes expressing perforin mRNA indicates in situ activation of lymphocytes within the epithelial compartment in florid coeliac disease upon ingestion of a gluten-containing diet in patients predisposed to coeliac disease. PMID- 15880761 TI - An inhibitor of cyclin-dependent kinase, stress-induced p21Waf-1/Cip-1, mediates hepatocyte mito-inhibition during the evolution of cirrhosis. AB - During the evolution of cirrhosis, there is a relative decrease in volume percentage of hepatocytes and a relative increase in biliary epithelial cells and myofibroblasts. This is recognized histopathologically as a ductular reaction and leads to gradual distortion of the normal hepatic architecture. The final or decompensated stage of cirrhosis is characterized by a further decline in hepatocyte proliferation and loss of functional liver mass that manifests clinically as ascites, encephalopathy, and other signs of liver failure. In this report, we tested the hypothesis that p21-mediated hepatocyte mito-inhibition accelerates the evolution of cirrhosis using an established mouse model of decompensated biliary cirrhosis, p21-deficient mice, and liver tissue from humans awaiting liver replacement. Despite the same insult of long-term (12-week) bile duct ligation, mice prone to decompensation showed significantly more oxidative stress and hepatocyte nuclear p21 expression, which resulted in less hepatocyte proliferation, an exaggerated ductular reaction, and more advanced disease compared with compensation-prone controls. Mice deficient in p21 were better able than wild-type controls to compensate for long-term bile duct ligation because of significantly greater hepatocyte proliferation, which led to a larger liver mass and less architectural distortion. Mito-inhibitory hepatocyte nuclear p21 expression in humans awaiting liver replacement directly correlated with pathological disease stage and model of end-stage liver disease scoring. In conclusion, stress-induced upregulation of hepatocyte p21 inhibits hepatocyte proliferation during the evolution of cirrhosis. These findings have implications for understanding the evolution of cirrhosis and associated carcinogenesis. Supplementary material for this article can be found on the HEPATOLOGY website (http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html). PMID- 15880762 TI - Demonstration of kynurenine aminotransferases I and II and characterization of kynurenic acid synthesis in cultured cerebral cortical neurons. AB - The present study characterizes the synthesis of kynurenic acid (KYNA) from exogenously added kynurenine and its regulation by extrinsic factors, in cultured cerebral cortical neurons and, for comparison, in astrocytes incubated under identical conditions. The neuronal culture showed positive immunostaining for both kynurenic acid aminotransferase (KAT) isoforms I and II. Neurons synthesized KYNA at a rate about 2.3 times higher than astrocytes. Neuronal, but not astrocytic, KYNA synthesis was lowered approximately 30% by ionotropic glutamate receptor agonists [(R,S)-3-hydroxy-5-methoxyloxasole-4-propionic acid (AMPA; 100 microM) and N-methyl-D-aspartic acid (NMDA; 100 microM)] and depolarizing agents [KCl (50 mM) and 4-aminopyridine (4-AP; 10 microM)]. Neuronal and astrocytic synthesis alike were vulnerable to inhibition exerted by the aminotransferase inhibitor aminooxyacetic acid (AOAA), glutamate (IC50: 31 and 85 microM, respectively), substrates of the L-amino transport system [leucine (Leu); IC50: 19 and 42 microM, respectively] and 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid (BCH; IC50: 19 and 28 microM, respectively). Glutamine (Gln), which is a metabolic precursor of glutamate in astrocytes and L-system substrate in both cell types, inhibited KYNA synthesis both in neurons and in astrocytes (IC50: 268 and 318 microM, respectively). alpha-Ketoisocaproic acid (KIC), a Leu transamination product that is produced mainly in astrocytes and shuttled to neurons to modulate intraneuronal concentration of glutamate, stimulated KYNA synthesis in neurons but did not affect the synthesis in astrocytes. In conclusion, this study is the first to demonstrate active, regulation-prone KYNA synthesis in neurons. PMID- 15880763 TI - Comparisons of aquatic versus terrestrial predatory strikes in the pitviper, Agkistrodon piscivorus. AB - Recent studies comparing terrestrial versus aquatic locomotion in animals have shown that changes in kinematics, muscular activation patterns, and performance across media are often dramatic. Surprisingly, however, despite the importance of feeding to the survival of most animals, few studies have compared differences in feeding behaviour between media. The present study compares prey-capture behaviour, strike success, strike velocity and acceleration, and prey-capture kinematics in a semi-aquatic pitviper (Agkistrodon piscivorus) when capturing both terrestrial (mice) and aquatic (fish) prey in a standardized laboratory setting. Strike velocity and acceleration did not differ significantly between media, but instead were positively correlated with initial prey distance. By contrast, the kinematics of terrestrial and aquatic strikes differed significantly in several aspects: max gape angle during the retraction phase, angular velocity of mouth closing during the strike, and the initial head angle before the strike. Terrestrial strikes were associated with higher gape angles during the retraction phase, higher angular velocities of mouth closure, and a more inclined head angle at the onset of the strike. Finally, strike success differed significantly between strike types, with terrestrial strikes being considerably more successful than aquatic strikes. Strike success likely differed due to the relatively slow mouth-closing velocity of aquatic strikes. PMID- 15880764 TI - D-lactate metabolism in starved Octopus ocellatus. AB - The concentrations of D- and L-lactate, methylglyoxal and pyruvate were measured in tissues of normal and starved Octopus ocellatus. D-Lactate was always more abundant than L-lactate in the tissues. D-Lactate, pyruvate and methylglyoxal were present in 320, 94 and 43 times higher concentrations in tentacle of O. ocellatus of control group than those in normal rat skeletal muscle. The D lactate concentration in the tentacle of O. ocellatus was 17-fold higher than that in Octopus vulgars. The activities of enzymes involved with D-lactate metabolism such as pyruvate kinase, octopine dehydrogenase, glyoxalase I and II and lactate dehydrogenase were measured in those tissues. The activities of glyoxalase I and II, and D-lactate dehydrogenase were increased in mantle and tentacle of starved octopus, while the levels of D-lactate and related metabolites were lowered in these tissues. The experimental results presented in this report and up to the present indicate that D-lactate is actively used for energy production in the tentacle and mantle of the starved animals. In octopus, especially starved octopus D-lactate was actively produced from methylglyoxal, which is formed via aminoacetone from threonine and glycine. PMID- 15880765 TI - Dynamics and histological characteristics of gonadal sex differentiation in pejerrey (Odontesthes bonariensis) at feminizing and masculinizing temperatures. AB - This study evaluated the effects of different temperatures on the histological process of sex differentiation in the pejerrey Odontesthes bonariensis, a fish with marked temperature-dependent sex determination (TSD), at feminizing, neutral, and masculinizing temperatures. Fish reared at three temperatures (17 degrees C, 24 degrees C, and 29 degrees C) from hatching were sampled weekly until 11 weeks and their gonads were examined by histology. The percentages of females at 17 degrees C, 24 degrees C, and 29 degrees C were 100%, 73%, and 0%, respectively. Sex differentiation occurred earlier and at a smaller body size at higher temperatures in both sexes. The first signs of ovarian differentiation were observed at 4 and 7 weeks at 24 degrees C and 17 degrees C, respectively, and those of testicular differentiation at 4 and 7 weeks at 29 degrees C and 24 degrees C, respectively. Body or gonadal growth rates before sex differentiation were not proportional to temperature and showed no sexual dimorphism at 24 degrees C, where both sexes were present. Thus, differential growth rate is probably not a factor in TSD or histological sex differentiation in pejerrey. Blood vessels were formed before sex differentiation in both sexes and at all temperatures, and may be important for sex differentiation. No signs of intersexuality were found in any of the groups, and this characterizes pejerrey as the differentiated type of gonochorist even at feminizing and masculinizing temperatures. Ovaries were formed by the same histological processes at feminizing (17 degrees C) and neutral (24 degrees C) temperatures and without any pathological features such as germ cell degeneration. The process of testicular formation was generally similar at 24 degrees C and 29 degrees C, but some fish at 29 degrees C had widespread germ cell degeneration before sex differentiation. This suggests that pathological processes leading to germ cell death, such as heat-induced dysfunction of the supporting somatic cells, could be involved in masculinization of the genetic females at high temperatures. PMID- 15880766 TI - Experimental analysis of nature-nurture interactions. AB - The presumed opposition of nature and nurture has been a major concern of western civilization since its beginnings. Christian theologians interpreted Adam and Eve's eating of the forbidden fruit as the origin of an inherited 'original sin'. Saint Augustine explicitly applied the concept to human mental development, arguing that, because of original sin, children are inclined toward evil and education requires physical punishment. For centuries, it was considered parents' moral and religious obligation, not to nurture their children, in our current sense of that word, but to beat the willfulness out of them. 16thC humanists fought back, arguing that "schools have become torture chambers" while it is adults "who corrupt young minds with evil". Locke's (1690) statement that children are born as a 'white paper' was crucial in rejecting the dogma of an inborn (and sinful) nature. The original sin vs. white paper argument merged with another ancient dichotomy: inborn instinct (which controls animal behavior) vs. the reason and free will which humans have. Darwin made the concept of inherited instinct, common to both man and animals, one cornerstone of his theory of evolution. The 20(th)C saw scientists recast the debate as instinct vs. learning, bitterly argued between behaviorists and ethologists. Laboratory experimentation and field observation showed that behavior could develop without learning but also that conditioning paradigms could powerfully mold behavior. The progress of genetics and neurobiology has led to the modern synthesis that neural development, and hence behavior, results from the interdependent action of both heredity and environment. PMID- 15880767 TI - Panorama Ab Microarray Cell Signaling kit: a unique tool for protein expression analysis. AB - Antibody arrays are a promising new tool for mass analysis of protein level changes in cells responding to different stimuli. Here we describe a novel antibody array system called Panorama Ab Microarray Cell Signaling, that contains 224 antibodies spotted on FAST nitrocellulose-coated slides that can detect protein levels as low as a few nanograms per mL. The antibodies spotted are specific for proteins important in various areas of cell signaling such as phosphorylation, cell cycle, apoptosis, nuclear signaling and cytoskeleton proteins. Furthermore, for some of the protein targes, the Panorama Ab Microarray can detect phosphorylated and nonphosphorylated forms of the traget protein. We found that treatment of the slides post-spotting is important for the array performance (ratio of signal to background) and its stability. Panorama Ab Microarray was used to analyze changes in protein expression in F9 embryonic carcinoma cells stimulated to differentiate by all-trans retinoic acid. We found that the level of several proteins, among them cell cycle regulators and kineases, was either up- or down-regulated. For more than ten protein targets, the results obtained by the Panorama Ab Microarray were confirmed by immunoblotting. PMID- 15880768 TI - United States National Heart, Lung, And Blood Institute Proteomics Initiative. PMID- 15880769 TI - Genetic basis for innovations in floral organ identity. AB - Of the many innovations associated with the radiation of the angiosperms, the evolution of a petal identity program is among the best understood from a genetic standpoint. Although the existing data do indicate that similar genetic mechanisms control petal development across diverse taxa, there is also considerable evidence for variability in petal identity programs, likely due to a number of factors. These points are illustrated through a review of our current knowledge on the subject, integrating phylogenetic, morphological, and genetic studies. Comparative studies of petal identity highlight the complex nature of homology in plants and stand as a cautionary tale for the interpretation of gene expression data. PMID- 15880771 TI - The digits of the wing of birds are 1, 2, and 3. A review. AB - Fossil evidence documenting the evolutionary transition from theropod dinosaurs to birds indicates unambiguously that the digits of the wing of birds are digits 1, 2, and 3. However, some embryological evidence suggests that these digits are 2, 3, and 4. This apparent lack of correspondence has been described as the greatest challenge to the widely accepted theropod-bird link (Zhou 2004. Naturwissenschaften 91:455-471). Here we review the pertinent literature regarding the debate on the origin of birds and wing digital identity and the evidence in favor of a 1, 2, 3 identity of the wing digits. Recent molecular evidence shows that the expression of Hoxd12 and Hoxd13 in the developing wing supports the theropod-bird link. In the chicken foot and in the mouse hand and foot, digit 1 is the only digit to combine the expression of Hoxd13 with the absence of expression of Hoxd12. The same is observed in the anterior digit of the wing, suggesting it is a digit 1, as expected for a theropod. Nevertheless, Galis et al. (2005. J Exp Zool (Mol Dev Evol) in press), argue that Hoxd12 and Hoxd13 expression patterns in mutant limbs do not allow distinguishing the most anterior digit in the bird wing from digit 2. They also argue that constraints to the evolution of limb development support the 2, 3, 4 identity of the wing digits. However, the case put forward by Galis et al. is biased and flawed with regard to interpretation of mutant limbs, developmental mechanisms, stages observed, and the description of the evolutionary variation of limb development. Importantly, Galis et al. do not present evidence from wild-type limbs that counters the conclusions of Vargas and Fallon (2005. J Exp Zool (Mol Dev Evol) 304B(1):85-89), and fail to provide molecular evidence to specifically support the hypothesis that the wing digits are 2, 3, and 4. The expression of Hoxd12 and Hoxd13 in the developing wing is consistent with the hypothesis that birds are living dinosaurs; this view can lead to a greater understanding of the actual limits to the evolutionary variation of limb development. PMID- 15880772 TI - Four types of Smad4 found in the common carp, Cyprinus carpio. AB - Smad4 is defined as the common-mediator Smad (Co-Smad) required for transducing signals for all transforming growth factor-beta (TGF-beta) superfamily members. In this study, we have isolated eight distinct Smad4 full-length cDNAs from the common carp (Cyprinus carpio). These cDNAs were classified into four types and each type consisted of two subtypes. The eight cDNAs encoded four distinct proteins ranging from 505aa to 568aa in size, with close similarities in the Mad homology 1 and 2 (MH1 and MH2, respectively), but with differences in the linker regions and the C-terminus as well as in the 5'- and 3'-untranslated regions. Genomic Southern blotting demonstrated the existence of at least six Smad4 gene loci in the carp genome, meaning that the multiple forms of the carp Smad4 cDNAs were not due to allelic variations. Reverse transcriptase polymerase chain reaction (RT-PCR)/Southern hybridizations showed different expression patterns among the four types of Smad4s. These results suggest that some of carp Smad4s have deviated from the original function of Smad4 through vertebrate evolution, and regulated the TGF-beta signaling pathway by changing the expression level in tissues. PMID- 15880773 TI - Hox genes, digit identities and the theropod/bird transition. AB - Vargas and Fallon (2005. J Exp Zool (Mol Dev Evol) 304B:86-90) propose that Hox gene expression patterns indicate that the most anterior digit in bird wings is homologous to digit 1 rather than to digit 2 in other amniotes. This interpretation is based on the presence of Hoxd13 expression in combination with the absence of Hoxd12 expression in the second digit condensation from which this digit develops (the first condensation is transiently present). This is a pattern that is similar to that in the developing digit 1 of the chicken foot and the mouse hand and foot. They have tested this new hypothesis by analysing Hoxd12 and Hoxd13 expression patterns in two polydactylous chicken mutants, Silkie and talpid2. They conclude that the data support the notion that the most anterior remaining digit of the bird wing is homologous to digit 1 in other amniotes either in a standard phylogenetic sense, or alternatively in a (limited) developmental sense in agreement with the Frameshift Hypothesis of Wagner and Gautier (1999, i.e., that the developmental pathway is homologous to the one that leads to a digit 1 identity in other amniotes, although it occurs in the second instead of the first digit condensation). We argue that the Hoxd12 and Hoxd13 expression patterns found for these and other limb mutants do not allow distinguishing between the hypothesis of Vargas and Fallon (2005. J Exp Zool (Mol Dev Evol) 304B:86-90) and the alternative one, i.e., the most anterior digit in bird wings is homologous to digit 2 in other amniotes, in a phylogenetic or developmental sense. Therefore, at the moment the data on limb mutants does not present a challenge to the hypothesis, based on other developmental data (Holmgren, 1955. Acta Zool 36:243-328; Hinchliffe, 1984. In: Hecht M, Ostrom JH, Viohl G, Wellnhofer P, editors. The beginnings of birds. Eichstatt: Freunde des Jura-Museum. p 141-147; Burke and Feduccia, 1997. Science 278:666-668; Kundrat et al., 2002. J Exp Zool (Mol Dev Evol) 294B:151-159; Larsson and Wagner, 2002. J Exp Zool (Mol Dev Evol) 294B:146-151; Feduccia and Nowicki, 2002. Naturwissenschaften 89:391-393), that the digits of bird wings are homologous to digits 2,3,4 in amniotes. We recommend further testing of the hypothesis by comparing Hoxd expression patterns in different taxa. PMID- 15880774 TI - Control of protein functional dynamics by peptide linkers. AB - Control of structural flexibility is essential for the proper functioning of a large number of proteins and multiprotein complexes. At the residue level, such flexibility occurs due to local relaxation of peptide bond angles whose cumulative effect may result in large changes in the secondary, tertiary or quaternary structures of protein molecules. Such flexibility, and its absence, most often depends on the nature of interdomain linkages formed by oligopeptides. Both flexible and relatively rigid peptide linkers are found in many multidomain proteins. Linkers are thought to control favorable and unfavorable interactions between adjacent domains by means of variable softness furnished by their primary sequence. Large-scale structural heterogeneity of multidomain proteins and their complexes, facilitated by soft peptide linkers, is now seen as the norm rather than the exception. Biophysical discoveries as well as computational algorithms and databases have reshaped our understanding of the often spectacular biomolecular dynamics enabled by soft linkers. Absence of such motion, as in so called molecular rulers, also has desirable functional effects in protein architecture. We review here the historic discovery and current understanding of the nature of domains and their linkers from a structural, computational, and biophysical point of view. A number of emerging applications, based on the current understanding of the structural properties of peptides, are presented in the context of domain fusion of synthetic multifunctional chimeric proteins. PMID- 15880775 TI - The influence of long-term chromatic adaptation on pigment cells and striped pigment patterns in the skin of the zebrafish, Danio rerio. AB - The striped pigment patterns in the flanks of zebrafish result from chromatophores deep within the dermis or hypodermis, while superficial melanophores associated with dermal scales add a dark tint to the dorsal coloration. The responses of these chromatophores were compared during the long term adaptation of zebrafish to a white or a black background. In superficial skin, melanophores, xanthophores, and two types of iridophores are distributed in a gradient along the dorso-ventral axis independent of the hypodermal pigment patterns. Within one week the superficial melanophores and iridophores changed their density and/or areas of distribution, which adopted the dorsal skin color and the hue of the flank to the background, but did not affect the striped pattern. The increases or decreases in superficial melanophores are thought to be caused by apoptosis or by differentiation, respectively. When the adaptation period was prolonged for more than several months, the striped color pattern was also affected by changes in the width of the black stripes. Some black stripes disappeared and interstripe areas were emphasized with a yellow color within one year on a white background. Such long-term alteration in the pigment pattern was caused by a decrease in the distribution of melanophores and a concomitant increase in xanthophores in the hypodermis. These results indicate that morphological responses of superficial chromatophores contribute to the effective and rapid background adaptation of dorsal skin and while prolonged adaptation also affects hypodermal chromatophores in the flank to alter the striped pigment patterns. PMID- 15880776 TI - Hyperglycaemic effects of 5-hydroxytryptamine and dopamine in the freshwater prawn, Macrobrachium malcolmsonii. AB - The effects of 5-hydroxytryptamine (5-HT; serotonin) and dopamine (DA) on tissue carbohydrate metabolism and haemolymph glucose levels in the freshwater prawn, Macrobrachium malcolmsonii, were investigated. Injection of 5-HT and DA produced hyperglycaemia in a dose-dependent and time-dependent manner, with DA being more effective than 5-HT. Interestingly, 5-HT and DA induced hyperglycaemia only in intact prawns but not in bilaterally eyestalk-ablated individuals. Total carbohydrate (TCHO) and glycogen levels decreased and phosphorylase activity increased in the hepatopancreas and muscle of intact prawns after being injected with 5-HT or DA. However, bilateral eyestalk ablation decreased haemolymph glucose and tissue phosphorylase activity and increased TCHO and glycogen levels of the hepatopancreas and muscle. Injection of 5-HT or DA did not cause significant changes in these variables in eyestalk-ablated prawns. It is hypothesized that 5-HT and DA induce hyperglycaemia in prawns by stimulating the release of crustacean hyperglycaemic hormone (CHH) from the X-organ sinus gland (XO-SG) complex located in the eyestalk. PMID- 15880777 TI - Changes in salinity and ionic compositions can act as environmental signals to induce a reduction in ammonia production in the African lungfish Protopterus dolloi. AB - The slender African lungfish, Protopterus dolloi, does not aestivate in a subterranean mud cocoon, but is capable of aestivating inside a layer of dried mucus on land during drought. In this study, we aimed to elucidate if a slight increase in salinity in association with changes in the ionic composition could act as signals for P. dolloi to decrease endogenous ammonia production, in preparation for aestivation when the external medium dries up. Specimens of P. dolloi exposed to 3 per thousand water for 6 days exhibited consistently lower daily urea excretion rate than the freshwater control. This led to significant decreases in the cumulative total nitrogenous wastes excreted on days 3, 5 and 6. On day 6, there were decreases in urea contents in various tissues and organs. Taken together, these results suggest that there was a decrease in the rate of urea synthesis, the magnitude of which was greater than the decrease in the rate of urea excretion, and therefore resulted in decreases in internal urea contents. A decrease in the rate of urea synthesis should result in a decrease in the rate of glutamine utilization, and subsequently led to the accumulations of glutamine and/or ammonia. However, there were no changes in contents of glutamine and ammonia in various tissues and organs in the experimental animals. A logical explanation for this is that there must be a simultaneous reduction in ammonia production; if not, ammonia would accumulate due to the decrease in rate of urea synthesis. Since fish were unfed during the experiment, endogenous ammonia must be derived mainly from amino acid catabolism. Therefore, these results suggest that a suppression of amino acid catabolism occurred in specimens exposed to 3 per thousand for 6 days. The differences in effects of freshwater and 3 per thousand water on endogenous ammonia production could not be due to food deprivation because both groups of fish were fasted for the same period. Because control and experimental fish were kept in water and because there were no changes in the wet mass of the fish and blood osmolality before and after the experiment, dehydration did not occur. Furthermore, both groups of fish have comparable blood pH, pO2 and pCO2 on day 6 as they had free access to air, and therefore CO2 retention could be eliminated as the initiating factor of suppressed endogenous ammonia production. In conclusion, our results suggest that P. dolloi could respond to increases in salinity and changes in ionic composition in the external medium by suppressing ammonia production in preparation for aestivation when the water dries up. PMID- 15880778 TI - The effect of environmental hypercapnia and salinity on the expression of NHE like isoforms in the gills of a euryhaline fish (Fundulus heteroclitus). AB - The current models for branchial acid excretion in fishes include Na(+)/H(+) exchange and the electrogenic excretion of H+ via H+-ATPase. The predominant route of acid excretion in some freshwater fishes is thought to be via the H+ ATPase/Na+ channel system. The euryhaline Fundulus heteroclitus may not fit this profile even when adapted to freshwater (FW). We hypothesize that the Na+/H+ exchanger (NHE) in this species may play a predominant role in acid-base regulation for both marine and FW adapted animals. Acidosis induced by ambient hypercapnia (1% CO2 in air), resulted in an increase in net H+ excretion to the water in F. heteroclitus pre-adapted to FW, brackish (isoosmotic; BW) and seawater (SW). Both FW and SW adapted mummichogs were tested for NHE protein expression using mammalian NHE antibodies, and we identified NHE-like immunoreactive proteins in gill membrane preparations from both groups. Hypercapnia induced a approximately three-fold elevation in gill NHE2-like protein in FW animals but SW adapted fish showed inconsistent NHE3-like protein expression. There was no change in NHE-1 levels in FW fish. In contrast, SW animals demonstrated a significant increase in both NHE1 and NHE3-like proteins following hypercapnia but limited expression of the NHE2 protein. We hypothesize that different isoforms of NHE may be preferentially expressed depending on the salinity to which the animals are adapted. Net H+ transfers during acidosis may be driven, at least in part by the action of these transporters. PMID- 15880779 TI - Design, construction, and characterization of a large synthetic human antibody phage display library. AB - Advances in proteomic research allow the identification of several hundred protein components in complex biological specimens. Structural information is typically lost during proteomic investigations. For this reason, the rapid isolation of monoclonal antibodies specific to proteins of interest would allow the study of structurally intact biological specimens, thus providing complementary proteomic information. Here, we describe the design, construction, characterization, and use of a large synthetic human antibody phage display library (ETH-2-Gold) containing three billion individual antibody clones. A large repertoire of antibodies with similar biochemical properties was produced by appending short variable complementarity-determining region 3 (CDR3) onto three antibody germline segments (DP47, DPK22, and DPL16), which are frequently found in human antibodies. The ETH-2-Gold library exhibits efficient display of antibody fragments on filamentous phage, as assessed by immunoblot. Furthermore, the library is highly functional, since >90% of clones express soluble antibodies in bacteria and since good quality monoclonal antibodies have been isolated against 16 different antigens. The usefulness of the library as a tool for generating monoclonal antibodies for biomedical applications was tested using the C-domain of tenascin-C (a marker of angiogenesis) as antigen and showing that specific antibodies to this target were able to stain vascular structures in tumor sections. PMID- 15880780 TI - Protein profile changes in the human breast cancer cell line MCF-7 in response to SEL1L gene induction. AB - The ectopic expression of the gene SEL1L in the human breast carcinoma cell line MCF-7 resulted in a reduction of the aggressive behaviour of these cells in vitro. In addition, in vivo analysis on a series of primary breast carcinomas revealed an association between the SEL1L protein levels and the patient's overall survival. We aimed to find those proteins, associated with SEL1L, which may be involved in modulating the aggressive or invasive behaviour of breast cancer cells. For this purpose, we used both the proteomic and microarray approaches. Image analysis of two-dimensional electropherograms revealed the presence of 27 qualitative and 35 quantitative variations between the MCF7-SEL1L expressing cells compared to control. Mass spectrometry identified 32 changing proteins mostly involved in cytoskeletal and metabolic activities, stress response and protein folding, selenoprotein synthesis and cellular proliferation. Five of these also showed changes in transcript levels, as assessed by Affymetrix microarray analysis. Interestingly, seven proteins: carbonic anhydrase (CA) II, ovarian/breast septin, S100A16 calcium binding protein, 14-3-3 protein sigma, proteasome subunit beta type 6, Hsp60 and protein disulphide-isomerase A3 merit particular attention since they are known to be involved in cancer, in response to cellular stress and in protein folding. PMID- 15880781 TI - First-principle studies of intermolecular and intramolecular catalysis of protonated cocaine. AB - We have performed a series of first-principles electronic structure calculations to examine the reaction pathways and the corresponding free energy barriers for the ester hydrolysis of protonated cocaine in its chair and boat conformations. The calculated free energy barriers for the benzoyl ester hydrolysis of protonated chair cocaine are close to the corresponding barriers calculated for the benzoyl ester hydrolysis of neutral cocaine. However, the free energy barrier calculated for the methyl ester hydrolysis of protonated cocaine in its chair conformation is significantly lower than for the methyl ester hydrolysis of neutral cocaine and for the dominant pathway of the benzoyl ester hydrolysis of protonated cocaine. The significant decrease of the free energy barrier, approximately 4 kcal/mol, is attributed to the intramolecular acid catalysis of the methyl ester hydrolysis of protonated cocaine, because the transition state structure is stabilized by the strong hydrogen bond between the carbonyl oxygen of the methyl ester moiety and the protonated tropane N. The relative magnitudes of the free energy barriers calculated for different pathways of the ester hydrolysis of protonated chair cocaine are consistent with the experimental kinetic data for cocaine hydrolysis under physiologic conditions. Similar intramolecular acid catalysis also occurs for the benzoyl ester hydrolysis of (protonated) boat cocaine in the physiologic condition, although the contribution of the intramolecular hydrogen bonding to transition state stabilization is negligible. Nonetheless, the predictability of the intramolecular hydrogen bonding could be useful in generating antibody-based catalysts that recruit cocaine to the boat conformation and an analog that elicited antibodies to approximate the protonated tropane N and the benzoyl O more closely than the natural boat conformer might increase the contribution from hydrogen bonding. Such a stable analog of the transition state for intramolecular catalysis of cocaine benzoyl-ester hydrolysis was synthesized and used to successfully elicit a number of anticocaine catalytic antibodies. PMID- 15880782 TI - All-electron density functional calculation on insulin with quasi-canonical localized orbitals. AB - An all-electron density functional (DF) calculation on insulin was performed by the Gaussian-based DF program, ProteinDF. Quasi-canonical localized orbitals (QCLOs) were used to improve the initial guess for the self-consistent field (SCF) calculation. All calculations were carried out by parallel computing on eight processors of an Itanium2 cluster (SGI Altix3700) with a theoretical peak performance of 41.6 GFlops. It took 35 h for the whole calculation. Insulin is a protein hormone consisting of two peptide chains linked by three disulfide bonds. The numbers of residues, atoms, electrons, orbitals, and auxiliary functions are 51, 790, 3078, 4439, and 8060, respectively. An all-electron DF calculation on insulin was successfully carried out, starting from connected QCLOs. Regardless of a large molecule with complicated topology, the differences in the total energy and the Mulliken atomic charge between initial and converged wavefunctions were very small. The calculation proceeded smoothly without any trial and error, suggesting that this is a promising method to obtain SCF convergence on large molecules such as proteins. PMID- 15880783 TI - Comparative study of lectin reactivity in the vomeronasal organ of human and nonhuman primates. AB - The main and accessory olfactory systems of certain mammals (e.g., rodents, ungulates, and carnivores) have been investigated using lectin histochemistry to probe for sugar residues that may reflect physiological aspects of signal transduction or development. Morphologically, the vomeronasal organs (VNOs) of strepsirrhine primates (lemurs and lorises) are typical of functional VNOs in other mammals, whereas in humans and chimpanzees the VNOs appear vestigial. However, the human VNO is considered functional by some authors. To elucidate the cellular nature of the VNO in human and chimpanzees, a panel of six lectins (Con A, ECL, PNA, RCA, s-WGA, and UEA-1) was applied to the VNO in eight species of primates, including humans and chimpanzees. The results indicated that there were few, if any, lectin-reactive cells in the human or chimpanzee VNO that resembled those seen in the vomeronasal neuroepithelium in other primates. The overall pattern of lectin reactivity in the human and chimpanzee VNO is unlike that seen in mammals with chemosensory VNOs, suggesting that the VNO of these hominoids does not function similarly to that of other primates. PMID- 15880784 TI - Differential effects of growth/differentiation factor 5 and glial cell line derived neurotrophic factor on dopaminergic neurons and astroglia in cultures of embryonic rat midbrain. AB - Parkinson's disease is characterized by the progressive degeneration of midbrain dopaminergic neurons. Several studies have examined the effects of the dopaminergic neurotrophins growth/differentiation factor 5 (GDF5) and glial cell line-derived neurotrophic factor (GDNF) on these neurons in vitro. However, there is little information regarding their effects on astroglial cells. Here, the effects of GDF5 and GDNF on dopaminergic neuronal and astroglial survival and differentiation in embryonic rat midbrain cultures were examined. Both GDF5 and GDNF enhanced the survival and differentiation of dopaminergic neurons. GDF5 significantly increased the survival of astroglial cells, whereas GDNF had no significant effect on these cells. The possible involvement of astroglia in the dopaminergic neurotrophic effect induced by GDF5 was investigated by examining the effect of GDF5 on the survival of dopaminergic neurons in glia-depleted midbrain cultures. There was no significant difference between the survival of dopaminergic neurons in glia-depleted cultures treated with GDF5 and that in mixed cell cultures treated with GDF5, suggesting that GDF5 acts directly on dopaminergic neurons in exerting its neurotrophic effect. GDF5 and GDNF have been established as potent neurotrophic factors for dopaminergic neurons. However, the effects of adding a combination of these neurotrophins to midbrain cultures have not been previously examined. The present study found that combined treatment with GDF5 and GDNF significantly increased the survival of dopaminergic neurons in cultures compared with that in cultures treated with either neurotrophin alone. This was an additive effect, indicating that these neurotrophins act on separate subpopulations of dopaminergic neurons. PMID- 15880785 TI - Different mechanisms preclude mutant CLDN14 proteins from forming tight junctions in vitro. AB - Mutations in claudin 14 (CLDN14) cause nonsyndromic DFNB29 deafness in humans. The analysis of a murine model indicated that this phenotype is associated with degeneration of hair cells, possibly due to cation overload. However, the mechanism linking these alterations to CLDN14 mutations is unknown. To investigate this mechanism, we compared the ability of wild-type and missense mutant CLDN14 to form tight junctions. Ectopic expression in L mouse fibroblasts (LM cells) of wild-type CLDN14 protein induced the formation of tight junctions, while both the c.254T>A (p.V85D) mutant, previously identified in a Pakistani family, and the c.301 G>A (p.G101R) mutant, identified in this study through the screen of 183 Spanish and Greek patients affected with sporadic nonsyndromic deafness, failed to form such junctions. However, the two mutant proteins differed in their ability to localize at the plasma membrane. We further identified hitherto undescribed exons of CLDN14 that are utilized in alternative spliced transcripts. We demonstrated that different mutations of CLDN14 impaired by different mechanisms the ability of the protein to form tight junctions. Our results indicate that the ability of CLDN14 to be recruited to these junctions is crucial for the hearing process. PMID- 15880786 TI - Causation in teratology-related litigation. PMID- 15880787 TI - Methamphetamine and lipid peroxidation in the rat retina. AB - BACKGROUND: The use of psychoactive drugs during adolescence and early adult life has increased in the last few decades. It is known that developmental exposure to psychostimulants affects the sensory systems, and the retina has been shown to be a target tissue. This work was conducted to evaluate the pattern of lipid peroxidation in the rat retina following prenatal exposure to methamphetamine (MA). METHODS: Pregnant female Wistar rats were given MA (5 mg/kg of body weight/day; SC, in 0.9% saline) from GD 8 to 22. Offspring were sacrificed at postnatal days (PNDs) 7, 14, and 21. The retinas were homogenized, and both the total antioxidant and superoxide dismutase (SOD) activities were measured by enzymatic-colorimetric methods. The lipid peroxidation byproducts (malondialdehyde [MDA] and MDA-like metabolites) were measured by the thiobarbituric acid test. RESULTS: Total antioxidant levels were lower in the MA group at PND 21 in both males and females. The activity of SOD was higher in PND 7 females from the MA group. MDA levels were higher in the MA group at PND 21 in both genders. CONCLUSIONS: These findings suggest that prenatal-induced MA toxicity in the retina may be related to lipid peroxidation processes and oxidative stress. PMID- 15880788 TI - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) inhibition of coronary vasculogenesis is mediated, in part, by reduced responsiveness to endogenous angiogenic stimuli, including vascular endothelial growth factor A (VEGF-A). AB - BACKGROUND: 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) exposure prior to chick embryo incubation (GD 0) induces dilated cardiomyopathy, and reduces myocardial hypoxia, vascular endothelial growth factor A (VEGF-A) expression, and coronary vascularization. We investigated whether reduced coronary vascularization 1) occurs in the absence of changes in cardiac morphology and 2) is associated with altered secretion of VEGF-A and/or an antivasculogenic factor. METHODS: Chicken eggs were treated with control (corn oil) or TCDD (0.075-0.3 pmol of TCDD/gm) on GD 5. In vivo cardiac morphology and artery number were determined on GD 10, while in vitro vascular outgrowth and VEGF-A secretion were determined from cardiac explants on GD 6. Effects of recombinant VEGF-A (rcVEGF-A), soluble flt-1 (sFlt-1) receptor plus rcVEGF-A, and control conditioned media were assessed in TCDD explants, while effects of TCDD-conditioned media was assessed in control explants. RESULTS: TCDD reduced coronary artery number in vivo by 53 +/- 8% and induced a dose-related reduction in tube outgrowth in vitro, but had no effect on cardiac morphology. All TCDD doses reduced explant VEGF-A secretion equally (43 +/- 3%), compared to control. sFlt-1 blocked outgrowth in control cultures and blocked rcVEGF-A-mediated rescue of outgrowth in TCDD explants. Control conditioned media partially rescued outgrowth from TCDD explants, while conditioned media from TCDD explants had no effect on controls. CONCLUSIONS: TCDD inhibition of coronary vascularization can occur in the absence of changes in cardiac morphology and is associated with reduced VEGF-A secretion but not an antivasculogenic factor. Since control media only partly rescues TCDD's inhibitory effect, we suggest that TCDD-exposed endothelial cells are less responsive to vasculogenic stimuli. PMID- 15880789 TI - An intervention study to increase knowledge and use of folic acid among relatives in neural tube defect-affected families in Washington, D.C. AB - BACKGROUND: Little is known about the level of knowledge and use of folic acid among near relatives in U.S. families of a child with spina bifida. We hypothesized that relatives would be more knowledgeable than the general population and more likely to take folic acid. Further, we hypothesized that relatives would be more motivated by an intervention to increase their use of folic acid. METHODS: We conducted an intervention study among females in families attending a hospital spina bifida clinic in Washington, DC. RESULTS: The 231 subjects consisted of the affected individuals, mothers, sisters, and aunts. The average age was 34 years. At baseline, most (87.4%) reported that they had heard of folic acid; 37.6% were currently taking multivitamins with folic acid and 6.9% were taking folic acid tablets. The intervention significantly increased both knowledge (to 99%) and intake of folic acid from 41.9 to 48.5%. Folic acid intake increased significantly among African-American women and women with less education, women who were older, married, with children, and nonsmokers. CONCLUSIONS: This intervention was successful in increasing folic acid intake among female relatives in spina bifida-affected families. By the end of the study, almost all women had heard of folic acid and folic acid use had increased by 16%. Among these women at higher than expected risk for having an affected child, this rate of intake, while more than the general population, still falls short of optimum. Fortification of food with folic acid may be the only way to ensure increased folic acid intake. PMID- 15880790 TI - Kinetic characterization of sequencing grade modified trypsin. AB - Prior to analysis by mass spectrometry, protein samples are often digested. Maximizing the peptide yield from digestion can increase the number of peptides detected and the confidence in protein identification. To determine the optimal conditions for digestion, the Michaelis-Menten kinetic parameters for Promega sequencing grade modified trypsin were measured over a range of temperatures and pHs. The results indicate that an increase in digestion temperature above 37 degrees C, the temperature traditionally used in digestion methods, could offer an increase in peptides detected. PMID- 15880791 TI - Ultra-high sensitivity multi-photon detection imaging in proteomics analyses. AB - We report on the use of 125I and 131I labeling and of new, multicolor, multi photon detection (MPD) methods to routinely and quantitatively detect protein spots on two-dimensional gel electrophoresis plates in the zeptomole to attomole range. We demonstrate that the MPD methodology can be used to detect radioactive labels on two-dimensional gels and has several characteristics that are advantageous for functional proteomics. First, by using single particle detectors, the sensitivity for detection of radiolabels can be improved dramatically. Second, because single particle detectors can differentiate the particle energies produced by different decay processes, it is possible to choose combinations of radioisotopes that can be detected and quantified individually on the same 2-D gel. Third, the MPD technology is essentially linear over six to seven orders of magnitude, i.e., it is possible to accurately quantify radiolabeled proteins over a range from at least 60 zeptomoles to 60 femtomoles. Finally for radionuclides that decay by electron capture, e.g., with emission of both beta and gamma rays, co-incident detection of two particles/photons can be used to detect such radionuclides well below background radiation levels. These methods are used to monitor acidic/phosphorylated proteins in as little as 60 ng of HeLa cells proteins. PMID- 15880793 TI - Antimicrobial peptides: new candidates in the fight against bacterial infections. AB - Antimicrobial peptides (AMPs) of innate origin are agents of the most ancient form of defense systems. They can be found in a wide variety of species ranging from bacteria through insects to humans. Through the course of evolution, host organisms developed arsenals of AMPs that protect them against a large variety of invading pathogens including both Gram-negative and Gram-positive bacteria. At a time of increasing bacterial resistance, AMPs have been the focus of investigation in a number of laboratories worldwide. Although recent studies show that some of the peptides are likely to have intracellular targets, the vast majority of AMPs appear to act by permeabilization of the bacterial cell membrane. Their activity and selectivity are governed by the physicochemical parameters of the peptide chains as well as the properties of the membrane system itself. In this review, we will summarize some of the recent developments that provide us with a better understanding of the mode of action of this unique family of antibacterial agents. Particular attention will be given to the determinants of AMP-lipid bilayer interactions as well as to the different pore formation mechanisms. The emphasis will be on linear AMPs but representatives of cysteine-bridged AMPs will also be discussed. PMID- 15880794 TI - Synthesis and conformational properties of protein fragments based on the Id family of DNA-binding and cell-differentiation inhibitors. AB - Id proteins are dominant negative regulators of the helix-loop-helix (HLH) transcription factors and are important during development, especially by preventing cell differentiation while inducing cell proliferation. In contrast, they are poorly expressed in healthy adults but are found in several tumor types. The Id HLH motif is responsible for the inhibitory activity, whereas not much is known about the role of the N- and C-termini. In the presented work, synthetic peptides reproducing the HLH, the N-terminal region, and the C-terminal region of the Id proteins were characterized by CD. The four HLH sequences built highly stable helical conformations, whereas the N- and C-termini were unstructured, with the exception of an alanine-rich fragment preceding the Id4 HLH motif. Deletion of the loop connecting the two helices led to helix destabilization for all four Id HLH peptides. In addition, modifications of the amino acid composition within the hydrophobic face of the helices of the Id1 HLH peptide induced conformational changes, mostly associated with loss of helix content. Moreover, a fragment containing the helix-2 and the C-terminus of the Id1 protein did not show any helical character. Therefore, both the helix propensity and stability of the HLH domain were shown to be strongly dependent on favorable interhelical contacts. In contrast, it is suggested that the regions beyond this domain could rather play a destabilizing role, for example, by increasing the flexibility of the folded protein. PMID- 15880795 TI - Late distortion of the original Palmaz stent implanted in postoperative lesions associated with congenital heart disease. AB - The objective of this study was to report late distortion of a Palmaz stent. Late distortion of an original Palmaz stent, implanted in an extracardiac lesion, is rare. We completed a 1-year follow-up of 54 patients who had been implanted with 80 Palmaz stents in extracardiac lesions. Distortion of two stents was detected in two patients. For case 1, we implanted a P188 stent for supravalvar pulmonary stenosis complicating an arterial switch operation in a 14-year-old girl. Seven months later, we found compression of the stent. Although we implanted two P308 stents anterior to the distorted stent, distortion of both stents developed after 1 month. Two more P308 stents placed inside each stent were gradually recompressed. A CAT scan showed compression of the stent by a dilated sinus of valsalva. For case 2, we implanted a P308 stent for stenosis of the superior vena cava after Williams operation in an 11-year-old boy. A chest X-ray documented longitudinal compression of the stent 27 months after implantation and a CAT scan showed the ascending aorta was in contact with the stent. A Palmaz stent may be distorted when implanted in a lesion adjacent to a pulsating aorta. PMID- 15880796 TI - Pharmacological induction of CFTR function in patients with cystic fibrosis: mutation-specific therapy. AB - CFTR mutations cause defects of CFTR protein production and function by different molecular mechanisms. Mutations can be classified according to the mechanisms by which they disrupt CFTR function. This understanding of the different molecular mechanisms of CFTR dysfunction provides the scientific basis for the development of targeted drugs for mutation-specific therapy of cystic fibrosis (CF). Class I mutations are nonsense mutations that result in the presence of a premature stop codon that leads to the production of unstable mRNA, or the release from the ribosome of a short, truncated protein that is not functional. Aminoglycoside antibiotics can suppress premature termination codons by disrupting translational fidelity and allowing the incorporation of an amino acid, thus permitting translation to continue to the normal termination of the transcript. Class II mutations cause impairment of CFTR processing and folding in the Golgi. As a result, the mutant CFTR is retained in the endoplasmic reticulum (ER) and eventually targeted for degradation by the quality control mechanisms. Chemical and molecular chaperones such as sodium-4-phenylbutyrate can stabilize protein structure, and allow it to escape from degradation in the ER and be transported to the cell membrane. Class III mutations disrupt the function of the regulatory domain. CFTR is resistant to phosphorylation or adenosine tri-phosphate (ATP) binding. CFTR activators such as alkylxanthines (CPX) and the flavonoid genistein can overcome affected ATP binding through direct binding to a nucleotide binding fold. In patients carrying class IV mutations, phosphorylation of CFTR results in reduced chloride transport. Increases in the overall cell surface content of these mutants might overcome the relative reduction in conductance. Alternatively, restoring native chloride pore characteristics pharmacologically might be effective. Activators of CFTR at the plasma membrane may function by promoting CFTR phosphorylation, by blocking CFTR dephosphorylation, by interacting directly with CFTR, and/or by modulation of CFTR protein-protein interactions. Class V mutations affect the splicing machinery and generate both aberrantly and correctly spliced transcripts, the levels of which vary among different patients and among different organs of the same patient. Splicing factors that promote exon inclusion or factors that promote exon skipping can promote increases of correctly spliced transcripts, depending on the molecular defect. Inconsistent results were reported regarding the required level of corrected or mutated CFTR that had to be reached in order to achieve normal function. PMID- 15880797 TI - Transseptal technique of percutaneous PFO closure results in persistent interatrial shunting. AB - Our goal was to review the effectiveness of the transseptal and tunnel techniques of patent foramen ovale (PFO) closure. Percutaneous PFO closure is an increasingly common treatment for prevention of paradoxical embolism and is typically performed by passing the device through the defect tunnel itself. The transseptal technique, in which the septum primum is punctured to create a hole through which the device is passed, has been proposed for patients with long tunnel PFO. From May 2001 to December 2003, 120 patients underwent PFO closure at our institution and were included in this analysis. Defect closure was assessed by transesophageal echocardiography (TEE) with bubble study. Clinical follow-up data were obtained by clinic visits or standardized telephone interviews. Device closure was successfully completed in all patients, with 12 (10%) undergoing transseptal closure and 108 (90%) undergoing tunnel closure. Immediately following the procedure, complete closure occurred in 6 of 12 (50%) of the transseptal group and 88 of 108 (81.5%) of the tunnel group (P = 0.0120). Of the 89 patients (74.2%) who returned for 6-month TEE, complete closure was demonstrated in 4 out of 10 (40%) of the transseptal group and 58 out of 79 (73.4%) of the tunnel group (P = 0.0303). There have been four transient ischemic attacks during a mean follow-up of 11 months. The transseptal technique of PFO closure results in a higher proportion of patients with persistent interatrial shunting when compared with the tunnel technique. Whether this is due to a difference in technique, device, or patient anatomy is unclear. PMID- 15880798 TI - Clinical outcomes after sirolimus-eluting stent implantation for de novo saphenous vein graft lesions. AB - The purpose of this study was to evaluate the clinical outcome of patients undergoing sirolimus-eluting stent implantation for de novo lesions within saphenous vein grafts (SVGs). Although the incidence of restenosis following sirolimus-eluting stenting (SES) of native coronary arteries is low, the efficacy of SES to treat de novo lesions within SVGs has not been well studied. A total of 35 patients underwent SES implantation of 39 lesions during 36 procedures. All patients had a minimum follow-up of 6 months following the index procedure. The mean bypass graft age was 10.1 +/- 6.5 years (range, 0-23 years). In-hospital major adverse cardiac events [death, myocardial infarction, thrombosis, or target vessel revascularization (TVR)] occurred in four patients (11%). Clinical follow up was obtained in 100% of patients (mean follow-up, 7.5 +/- 2.2 months). There was one cardiac death, presumed due to stent thrombosis. TVR occurred in only two patients (6%). Myocardial infarction (MI) occurred in four patients (11%), all attributable to a nontarget vessel. The combined endpoint of death, MI, or TVR occurred in seven patients (20%). Freedom from death, nonfatal MI, thrombosis, or any revascularization was 65%. Early experience indicates sirolimus-eluting stents for de novo saphenous vein graft lesions have a low (6%) rate of clinically driven target vessel revascularization. By 7-month follow-up, event free survival is limited primarily by disease in nontarget vessels. PMID- 15880799 TI - Fertilization. PMID- 15880800 TI - Two generation reproduction study of styrene by inhalation in Crl-CD rats. AB - This study was conducted to evaluate the potential adverse effects of styrene on reproductive capability from whole-body inhalation exposure of F0 and F1 parental animals. Assessments included gonadal function, estrous cyclicity, mating behavior, conception rate, gestation, parturition, lactation, and weaning in the F0 and F1 generations, and F1 generation offspring growth and development. Four groups of male and female Crl:CD(SD)IGS BR rats (25/sex/group) were exposed to 0, 50, 150, and 500 ppm styrene for 6 hr daily for at least 70 consecutive days prior to mating for the F0 and F1 generations. Inhalation exposure for the F0 and F1 females continued throughout mating and gestation through gestation day 20. Inhalation exposure of the F0 and F1 females was suspended from gestation day 21 through lactation day 4. On lactation days 1 through 4, the F0 and F1 females received styrene in virgin olive oil via oral gavage at dose levels of 66, 117, and 300 mg/kg/day (divided into three equal doses, approximately 2 hr apart). These oral dosages were calculated to provide similar maternal blood peak concentrations as provided by the inhalation exposures. Inhalation exposure of the F0 and F1 females was re-initiated on lactation day 5. Styrene exposure did not affect survival or clinical observations. Rats in the 150- and 500-ppm groups in both parental generations gained weight more slowly than the controls. There were no indications of adverse effects on reproductive performance in either the F0 or F1 generation. Male and female mating and fertility indices, pre-coital intervals, spermatogenic endpoints, reproductive organ weights, lengths of estrous cycle and gestation, live litter size and postnatal survival were similar in all exposure groups. Additionally, ovarian follicle counts and corpora lutea counts for the F1 females in the high-exposure group were similar to the control values. No adverse exposure-related macroscopic pathology was noted at any exposure level in the F0 and F1 generations. A previously characterized pattern of degeneration of the olfactory epithelium that lines the dorsal septum and dorsal and medial aspects of the nasal turbinates occurred in the F0 and F1 generation animals from the 500-ppm group. In the 500-ppm group, F2 birthweights were reduced compared to the control and F2 offspring from both the 150- and 500 ppm exposure groups gained weight more slowly than the controls. Based on the results of this study, an exposure level of 50 ppm was considered to be the NOAEL for F0 and F1 parental systemic toxicity; the NOAEL for F0 and F1 reproductive toxicity was 500 ppm or greater. PMID- 15880801 TI - Developmental neurotoxicity study of styrene by inhalation in Crl-CD rats. AB - This study was conducted to assess potential adverse functional and/or morphological effects of styrene on the neurological system in the F2 offspring following F0 and F1 generation whole-body inhalation exposures. Four groups of male and female Crl:CD (SD)IGS BR rats (25/sex/group) were exposed to 0, 50, 150, and 500 ppm styrene for 6 hr daily for at least 70 consecutive days prior to mating for the F0 and F1 generations. Inhalation exposure continued for the F0 and F1 females throughout mating and through gestation day 20. On lactation days 1 through 4, the F0 and F1 females received styrene in virgin olive oil via oral gavage at dose levels of 66, 117, and 300 mg/kg/day (divided into three equal doses, approximately 2 hr apart). Inhalation exposure of the F0 and F1 females was re-initiated on lactation day 5 and continued through weaning of the F1 or F2 pups on postnatal day (PND) 21. Developmental landmarks were assessed in F1 and F2 offspring. The neurological development of randomly selected pups from the F2 generation was assessed by functional observational battery, locomotor activity, acoustic startle response, learning and memory evaluations, brain weights and dimension measurements, and brain morphometric and histologic evaluation. Styrene exposure did not affect survival or the clinical condition of the animals. As expected from previous studies, slight body weight and histopathologic effects on the nasal olfactory epithelium were found in F0 and F1 rats exposed to 500 ppm and, to a lesser extent, 150 ppm. There were no indications of adverse effects on reproductive performance in either the F0 or F1 generation. There were exposure related reductions in mean body weights of the F1 and F2 offspring from the mid and high-exposure groups and an overall pattern of slightly delayed development evident in the F2 offspring only from the 500-ppm group. This developmental delay included reduced body weight (which continued through day 70) and slightly delayed acquisition of some physical landmarks of development. Styrene exposure of the F0 and F1 animals had no effect on survival, the clinical condition or necropsy findings of the F2 animals. Functional observational battery evaluations conducted for all F1 dams during the gestation and lactation periods and for the F2 offspring were unaffected by styrene exposure. Swimming ability as determined by straight channel escape times measured on PND 24 were increased, and reduced grip strength values were evident for both sexes on PND 45 and 60 in the 500-ppm group compared to controls. There were no other parental exposure-related findings in the F2 pre-weaning and post-weaning functional observational battery assessments, the PND 20 and PND 60 auditory startle habituation parameters, in endpoints of learning and memory performance (escape times and errors) in the Biel water maze task at either testing age, or in activity levels measured on PND 61 in the 500-ppm group. Taken together, the exposure-related developmental and neuromotor changes identified in F2 pups from dams exposed to 500 ppm occurred in endpoints known to be both age- and weight-sensitive parameters, and were observed in the absence of any other remarkable indicators of neurobehavioral toxicity. Based on the results of this study, an exposure level of 50 ppm was considered to be the NOAEL for growth of F2 offspring; an exposure level of 500 ppm was considered to be the NOAEL for F2 developmental neurotoxicity. PMID- 15880802 TI - Theoretical studies on the electronic and optical properties of two new alternating fluorene/carbazole copolymers. AB - Poly(fluorene)-type materials are widely used in polymer-based emitting devices. During operation there appears, however, an additional emission peak at around 2.3 eV, leading to both a color instability and reduced efficiency. The incorporation of the carbazole units has been proven to efficiently suppress the keto defect emission. In this contribution, we apply quantum-chemical techniques to investigate two series of alternating fluorene/carbazole oligomers and copolymers poly[2,7-(N-(2-methyl)-carbazole)-co-alt-2,7-m(9,9-dimethylfluorene)], namely, PFmCz (m = 1,2) and gain a detailed understanding of the influence of carbazole units on the electronic and optical properties of fluorene derivatives. The electronic properties of the neutral molecules, HOMO-LUMO gaps (Delta(H-L)), in addition to the positive and negative ions, are studied using B3LYP functional. The lowest excitation energies (E(g)s) and the maximal absorption wavelength lambda(abs) of PFmCz (m = 1,2) are studied, employing the time dependent density functional theory (TD-DFT). The properties of the two copolymers, such as Delta(H-L), E(g), IPs, and EAs were obtained by extrapolating those of the oligomers to the inverse chain length equal to zero (1/n = 0). The outcomes showed that the carbazole unit is a good electron-donating moiety for electronic materials, and the incorporation of carbazole into the polyfluorene (PF) backbone resulted in a broadened energy gap and a blue shift of both the absorption and photoluminescence emission peaks. Most importantly, the HOMO energies of PF1Cz and PF2Cz are both a higher average (0.4 eV) than polyfluorene (PF), which directly results in the decreasing of IPs of about 0.2 eV more than PF, indicating that the carbazole units have significantly improved the hole injection properties of the copolymers. In addition, the energy gap tends to broaden and the absorption and emission peaks are gradually blue-shifted to shorter wavelengths with an increase in the carbazole content in the copolymers. This is due to the interruption of the longer conjugation length of the backbone in the (F1Cz)(n) series. PMID- 15880803 TI - DFT study and Monte Carlo simulation on proton transfers of 2-amino-2-oxazoline, 2-amino-2-thiazoline, and 2-amino-2-imidazoline in the gas phase and in water. AB - Density functional theory (DFT) and Monte Carlo (MC) simulation with free energy perturbation (FEP) techniques have been used to study the tautomeric proton transfer reaction of 2-amino-2-oxazoline, 2-amino-2-thiazoline, and 2-amino-2 imidazoline in the gas phase and in water. Two reaction pathways were considered: the direct and water-assisted transfers. The optimized structures and thermodynamic properties of stationary points for the title reaction system in the gas phase were calculated at the B3LYP/6-311+G(d, p) level of theory. The potential energy profiles along the minimum energy path in the gas phase and in water were obtained. The study of the solvent effect of water on the proton transfer of 2-amino-2-oxozoline, 2-amino-2-thiazoline, and 2-amino-2-imidazoline indicates that water as a solvent is favorable for the water-assisted process and slows down the rate of the direct transfer pathway. PMID- 15880804 TI - Spot overlapping in two-dimensional maps: a serious problem ignored for much too long. AB - In the analysis of a neuroblastoma xenograft implanted in mice using two dimensional maps, some 85 proteins were found to be up- or down-regulated (out of a total of 264 detected by a medium-sensitivity colloidal Coomassie stain). When these spots were eluted and analysed by mass spectrometry in a quadrupole time of flight mass spectrometer, a number of spots were found to be envelopes of different polypeptide chains. Out of a total of 74 proteins identified, 52 (71%) were found to be singlets, 14 (19%) were doublets, 6 (8%) were triplets, 1 was a quadruplet and 1 a quintuplet. Analysis of the DeltapI and DeltaMr of all species contained in a single gel segment eluted helped point out potential errors in protein identification. This was a unique case, in that very minute bioptic sample loads were applied to the gel. In normal cases, where sample loads of ca. 1 mg of total protein are applied and typically at least 1000 spots are visualised, the singlets will be the minority, rarely exceeding 30% of all spots analysed. The experimental data on the abundance of overlapping spots were in excellent agreement with theoretical data calculated on the basis of the statistical theory of spot overlapping, originally proposed by Davis and further developed by some of the authors. Ways and means for minimizing spot overlap and visualising a greater number of spots in a two-dimensional map are discussed. PMID- 15880805 TI - Linking angiogenesis to bone destruction in arthritis. PMID- 15880806 TI - Evidence for a role of the genomic region of the gene encoding for the alpha1 chain of type IX collagen (COL9A1) in hip osteoarthritis: A population-based study. AB - OBJECTIVE: Type IX collagen proteoglycan is an important protein in collagen networks and has been implicated in hip osteoarthritis (OA). We studied 2 COL9A1 markers (509-8B2 and 509-12B1) in relation to radiographic OA, within the framework of the Rotterdam Study, a population-based study of 7,983 subjects ages 55 years and older. METHODS: We used 2 different designs, as follows: 1) a linkage study of 83 probands with multiple joints affected with radiographic OA and their 221 siblings, yielding 445 sibpairs who participated in the study, and 2) an association study in a series of 71 patients with radiographic hip OA and 269 controls without radiographic OA. All subjects were characterized for the 2 COL9A1 markers, 509-8B2 and 509-12B1. The mean test was used to assess the proportion of alleles shared in concordantly affected and unaffected sibpairs. The chi-square test was used to compare the allele distributions in patients and controls. RESULTS: Affected sibpairs with radiographic hip OA shared alleles identical by descent at markers 8B2 and 12B1 significantly more often than expected (mean +/- SD 0.66 +/- 0.07 and 0.65 +/- 0.08, respectively; P < 0.05). No excess sharing for radiographic OA was observed at other joint sites. When comparing the frequency of marker 8B2 and 12B1 alleles in subjects with radiographic OA and controls, the frequency of 8B2 alleles in subjects with radiographic OA differed significantly(P = 0.01) from that in controls. CONCLUSION: Our data suggest that susceptibility for hip OA is conferred within or close to the COL9A1 gene in linkage disequilibrium with the COL9A1 509-8B2 marker. PMID- 15880807 TI - Gene expression profiling of minor salivary glands clearly distinguishes primary Sjogren's syndrome patients from healthy control subjects. AB - OBJECTIVE: To identify gene expression signatures in minor salivary glands (MSGs) from patients with primary Sjogren's syndrome (SS). METHODS: A 16K complementary DNA microarray was used to generate gene expression profiles in MSGs obtained from 10 patients with primary SS and 10 control subjects. The data were analyzed by 2 different strategies, one strict primary analysis and one subanalysis that allowed for inclusion of genes with no signal in more than 3 samples from each group. The results were validated by quantitative reverse transcriptase polymerase chain reaction techniques. RESULTS: We found a distinct difference in gene expression levels in MSGs, enabling a simple class prediction method to correctly classify 19 of the 20 samples as either patient or control, based on the top 5 differentially expressed genes. The 50 most differentially expressed genes in the primary SS group compared with the control group were all up regulated, and a clear pattern of genes involved in chronic inflammation was found. CXCL13 and CD3D were expressed in >/=90% of primary SS patients and in /=12 weeks of MTX therapy were randomly assigned to receive weekly IM gold or placebo in addition to MTX. Gold was administered according to a standard protocol developed for the study. The primary outcome measure was the percentage of patients who met the American College of Rheumatology (ACR) 20% improvement criteria (achieved an ACR20 response) at week 48. Secondary outcomes included the percentages of patients achieving ACR50 and ACR70 responses, the individual criteria that make up the primary outcome, quality of life, direct and indirect health care costs, intraarticular steroid use, and adverse events, among other measures. Statistical analyses were based on an intent-to-treat strategy. RESULTS: Sixty-one percent of patients receiving gold achieved an ACR20 response compared with 30% of patients receiving placebo (chi(2) = 6.04, P = 0.014; logistic regression odds ratio 3.64 [95% confidence interval 1.3, 10.4], P = 0.016). Twenty-six percent of patients receiving gold achieved an ACR50 response compared with 4% of patients receiving placebo (P = 0.017), and 21% of patients receiving gold achieved an ACR70 response compared with 0% of patients receiving placebo (P = 0.011). From both clinical and cost-effectiveness perspectives, gold was the preferred and dominant strategy. Study treatment was discontinued in 23 patients (14 in the placebo group compared with 9 in the gold group; P = 0.022) due to loss to followup, adverse events, or lack of efficacy. CONCLUSION: In RA patients with a suboptimal response to MTX, adding weekly IM gold causes significant clinical improvement. Adverse events were minor, and IM gold-related adverse events led to discontinuation in only 11% of the gold group over 48 weeks. PMID- 15880811 TI - Translating osteoarthritis into numbers: the importance of alignment in knee radiography for clinical trials of structure-modifying drugs. PMID- 15880812 TI - ADAMTS-9 is synergistically induced by interleukin-1beta and tumor necrosis factor alpha in OUMS-27 chondrosarcoma cells and in human chondrocytes. AB - OBJECTIVE: To compare induction of the aggrecanases (ADAMTS-1, ADAMTS-4, ADAMTS 5, ADAMTS-8, ADAMTS-9, and ADAMTS-15) by interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) in chondrocyte-like OUMS-27 cells and human chondrocytes, and to determine the mechanism of induction of the most responsive aggrecanase gene. METHODS: OUMS-27 cells were stimulated for different periods of time and with various concentrations of IL-1beta and/or TNFalpha. Human chondrocytes obtained from osteoarthritic joints and human skin fibroblasts were also stimulated with IL-1beta and/or TNFalpha. Total RNA was extracted, reverse transcribed, and analyzed by quantitative real-time polymerase chain reaction and Northern blotting. ADAMTS-9 protein was examined by Western blotting, and the role of the MAPK signaling pathway for ADAMTS9 induction in IL-1beta-stimulated OUMS-27 cells was investigated. RESULTS: IL-1beta increased messenger RNA (mRNA) levels of ADAMTS4, ADAMTS5, and ADAMTS9 but not ADAMTS1 and ADAMTS8. The fold increase for ADAMTS9 mRNA was greater than that for mRNA of the other aggrecanase genes. The increase of ADAMTS9 mRNA by IL-1beta stimulation was greater in chondrocytes than in fibroblasts. The combination of IL-1beta and TNFalpha had a synergistic effect, resulting in a considerable elevation in the level of ADAMTS9 mRNA. ADAMTS-9 protein was also induced in IL-1beta-stimulated OUMS-27 cells. The MAPK inhibitors SB203580 and PD98059 decreased ADAMTS9 up-regulation in OUMS-27 cells. CONCLUSION: ADAMTS9 is an IL-1beta- and TNFalpha-inducible gene that appears to be more responsive to these proinflammatory cytokines than are other aggrecanase genes. Furthermore, these cytokines had a synergistic effect on ADAMTS9. Together with the known ability of ADAMTS-9 to proteolytically degrade aggrecan and its potential to cleave other cartilage molecules, the data suggest that ADAMTS-9 may have a pathologic role in arthritis. PMID- 15880813 TI - Decrease over time in the incidence of systemic rheumatoid vasculitis: Comment on the article by Turesson et al. PMID- 15880814 TI - MSight: an image analysis software for liquid chromatography-mass spectrometry. AB - Images obtained from high-throughput mass spectrometry (MS) contain information that remains hidden when looking at a single spectrum at a time. Image processing of liquid chromatography-MS datasets can be extremely useful for quality control, experimental monitoring and knowledge extraction. The importance of imaging in differential analysis of proteomic experiments has already been established through two-dimensional gels and can now be foreseen with MS images. We present MSight, a new software designed to construct and manipulate MS images, as well as to facilitate their analysis and comparison. PMID- 15880815 TI - Oxidative addition of the ethane C-C bond to Pd. An ab initio benchmark and DFT validation study. AB - We have computed a state-of-the-art benchmark potential energy surface (PES) for the archetypal oxidative addition of the ethane C-C bond to the palladium atom and have used this to evaluate the performance of 24 popular density functionals, covering LDA, GGA, meta-GGA, and hybrid density functionals, for describing this reaction. The ab initio benchmark is obtained by exploring the PES using a hierarchical series of ab initio methods [HF, MP2, CCSD, CCSD(T)] in combination with a hierarchical series of five Gaussian-type basis sets, up to g polarization. Relativistic effects are taken into account either through a relativistic effective core potential for palladium or through a full four component all-electron approach. Our best estimate of kinetic and thermodynamic parameters is -10.8 (-11.3) kcal/mol for the formation of the reactant complex, 19.4 (17.1) kcal/mol for the activation energy relative to the separate reactants, and -4.5 (-6.8) kcal/mol for the reaction energy (zero-point vibrational energy-corrected values in parentheses). Our work highlights the importance of sufficient higher angular momentum polarization functions for correctly describing metal-d-electron correlation. Best overall agreement with our ab initio benchmark is obtained by functionals from all three categories, GGA, meta-GGA, and hybrid DFT, with mean absolute errors of 1.5 to 2.5 kcal/mol and errors in activation energies ranging from -0.2 to -3.2 kcal/mol. Interestingly, the well-known BLYP functional compares very reasonably with a slight underestimation of the overall barrier by -0.9 kcal/mol. For comparison, with B3LYP we arrive at an overestimation of the overall barrier by 5.8 kcal/mol. On the other hand, B3LYP performs excellently for the central barrier (i.e., relative to the reactant complex) which it underestimates by only -0.1 kcal/mol. PMID- 15880816 TI - Autoantibodies to a 140-kd polypeptide, CADM-140, in Japanese patients with clinically amyopathic dermatomyositis. AB - OBJECTIVE: To identify novel autoantibodies specific for dermatomyositis (DM), especially those specific for clinically amyopathic DM (C-ADM). METHODS: Autoantibodies were analyzed by immunoprecipitation in 298 serum samples from patients with various connective tissue diseases (CTDs) or idiopathic pulmonary fibrosis (IPF). Antigen specificity of the sera was further examined by immunoblotting and indirect immunofluorescence (IF). The disease specificity and clinical features associated with the antibody of interest were determined. RESULTS: Eight sera recognized a polypeptide of approximately 140 kd (CADM-140 autoantigen) by immunoprecipitation and immunoblotting. Immunoreactivity was detected in the cytoplasm, and indirect IF revealed a granular or reticular pattern. Anti-CADM-140 antibodies were detected in 8 of 42 patients with DM, but not in patients with other CTDs or IPF. Interestingly, all 8 patients with anti CADM-140 antibodies had C-ADM. Among 42 patients with DM, those with anti-CADM 140 autoantibodies had significantly more rapidly progressive interstitial lung disease (ILD) when compared with patients without anti-CADM-140 autoantibodies (50% versus 6%; P = 0.008). CONCLUSION: These results indicate that the presence of anti-CADM-140 autoantibodies may be a novel marker for C-ADM. Further attention should be directed to the detection of rapidly progressive ILD in those patients with anti-CADM-140 autoantibodies. PMID- 15880817 TI - Gene therapy targeting the Tie2 function ameliorates collagen-induced arthritis and protects against bone destruction. AB - OBJECTIVE: In a previous study, we demonstrated that Tie2 regulates angiogenesis in arthritis. The current study was performed to determine whether systemic delivery of a soluble Tie2 receptor (ExTek) using an adenoviral vector (AdExTek) as a Tie2 inhibitor affects arthritis development and progression in an animal model. METHODS: We used a collagen-induced arthritis (CIA) mouse model to study the outcome of treatment with either AdExTek or a control vector. The onset, incidence, and severity of arthritis were quantified. Immunohistologic analysis of endothelium obtained from the paws was performed. Bone destruction in paws was analyzed using phase-contrast radiography. RESULTS: The data showed that systemic delivery of ExTek before disease development significantly inhibited the onset, incidence, and severity of arthritis. When AdExTek was given after disease onset, the severity of disease in mice treated with AdExTek was significantly lower than that in the control group at 35 days postimmunization, which correlated with significantly diminished angiogenesis in mouse paws. Strikingly, AdExTek treatment protected bone from erosion in the CIA model and reduced levels of RANKL. No differences in collagen-specific antibodies were detected between these 2 groups. CONCLUSION: We demonstrated that blocking Tie2 receptor activation inhibits angiogenesis and arthritis development and protects against bone destruction in a CIA mouse model. These findings identify Tie2 as a therapeutic target for arthritis treatment and imply that interventions designed to target the Tie2 pathway could be clinically beneficial. PMID- 15880818 TI - Reversal of the immunosuppressive properties of mesenchymal stem cells by tumor necrosis factor alpha in collagen-induced arthritis. AB - OBJECTIVE: Adult mesenchymal stem cells (MSCs) represent promising tools for therapeutic applications such as tissue engineering and cellular therapy. Recent data suggest that, due to their immunosuppressive nature, MSCs may be of interest to enhance allogeneic hematopoietic engraftment and prevent graft-versus-host disease. Using a murine model of rheumatoid arthritis (RA), this study investigated whether the immunosuppressive properties of MSCs could be of therapeutic value to inhibit reactive T cells in autoimmune diseases such as RA. METHODS: In mice with collagen-induced arthritis (CIA), we injected various doses of C3 MSCs at the time of immunization or booster injection, and subsequently evaluated the clinical and immunologic parameters. The immunosuppressive properties of MSCs were determined in vitro in mixed lymphocyte reactions with or without the addition of tumor necrosis factor alpha (TNFalpha). RESULTS: In the CIA model of arthritis, MSCs did not confer any benefit. Both the clinical and the immunologic findings suggested that MSCs were associated with accentuation of the Th1 response. Using luciferase-expressing MSCs, we were unable to detect labeled cells in the articular environment of the knee, suggesting that worsening of the symptoms was unlikely due to the homing of MSCs in the joints. Experiments in vitro showed that the addition of TNFalpha was sufficient to reverse the immunosuppressive effect of MSCs on T cell proliferation, and this observation was associated with an increase in interleukin-6 secretion. CONCLUSION: Our data suggest that environmental parameters, in particular those related to inflammation, may influence the immunosuppressive properties of MSCs. PMID- 15880819 TI - Development of proteoglycan-induced arthritis is critically dependent on Fcgamma receptor type III expression. AB - OBJECTIVE: To study the necessity for activating Fcgamma receptor types I and III (FcgammaRI and FcgammaRIII) in proteoglycan-induced arthritis (PGIA), a murine model of rheumatoid arthritis, and to determine whether usage of FcgammaRI or FcgammaRIII correlates with the Th1 phenotype or the autoantibody isotype in PGIA. METHODS: PGIA was induced by immunizing FcgammaRI(-/-), FcgammaRIII(-/-), and wild-type (WT) littermate mice with human PG. The development and severity of arthritis were monitored over time. PG-specific T cell interleukin-2 (IL-2) production and B cell antibody responses were assessed. FcgammaRIII blocking antibodies were used to inhibit arthritis in an adoptive transfer system. Inflammation in the hind paws was evaluated by assessing cytokine and chemokine messenger RNA (mRNA) transcripts by real-time polymerase chain reaction. RESULTS: FcgammaRI(-/-) mice developed arthritis with similar kinetics and severity as WT littermate controls, whereas FcgammaRIII(-/-) mice failed to develop the disease. Both FcgammaRI(-/-) and FcgammaRIII(-/-) mice produced similar amounts of PG specific antibody and IL-2 as littermate controls. Transfer of arthritis was successfully blocked in mice treated with a blocking antibody against FcgammaRIII. FcgammaRIII(-/-) mice displayed a significant decrease in cytokine and chemokine mRNA transcripts obtained from the hind paws of immunized mice, whereas FcgammaRI(-/-) mice demonstrated a similar increase in cytokine and chemokine transcripts as controls. CONCLUSION: These results demonstrate that FcgammaRIII expression is critical to the development of PGIA, and usage of FcgammaRIII correlates with the IgG1 isotype of the PG-specific antibody response. FcgammaRIII expression appears to be important in the effector phase of arthritis, possibly by activating cytokine- and chemokine-secreting cells in the joint. PMID- 15880820 TI - Pharmacogenetics and rheumatology: Molecular mechanisms contributing to variability in drug response. PMID- 15880821 TI - T cell costimulation by fractalkine-expressing synoviocytes in rheumatoid arthritis. AB - OBJECTIVE: Patients with rheumatoid arthritis (RA) accumulate prematurely aged T cells that have acquired a new profile of regulatory receptors. Many of the de novo-expressed receptors are typically found on natural killer cells, including CX(3)CR1, the receptor for the chemokine fractalkine (FKN). This study explored whether interactions between CX(3)CR1 and FKN are relevant for T cell functions in rheumatoid synovitis. METHODS: FKN expression was examined by real-time polymerase chain reaction and immunohistochemistry. CX(3)CR1 expression on peripheral blood T cells was analyzed by flow cytometry. T cell activation was quantified by determining proliferative responses, interferon-gamma (IFNgamma) secretion, and granule release. Fibroblast-like synoviocyte (FLS)/T cell adhesion was measured by the retention of 5-carboxyfluorescein diacetate succinimidyl ester-labeled T cells on FLS monolayers. RESULTS: FKN was expressed on cultured synovial fibroblasts and hyperplastic synoviocytes in the rheumatoid tissue. Among CD4+ T cells, only senescent CD28- T cells were positive for CX(3)CR1 (P < 0.001). Such CD4+,CD28-,CX(3)CR1+ T cells strongly adhered to FLS, with soluble FKN blocking the interaction. FKN expressed on FLS costimulated T cell-activating signals and amplified proliferation, IFNgamma production, and expulsion of cytoplasmic granules. CONCLUSION: Senescent CD4+ T cells that accumulate in rheumatoid arthritis aberrantly express CX(3)CR1. FKN, which is membrane-anchored on synoviocytes, enhances CD4+ T cell adhesion, provides survival signals, and costimulates the production of proinflammatory cytokines as well as the release of granules. By virtue of their altered receptor profile, senescent CD4+ T cells receive strong stimulatory signals from nonprofessional antigen-presenting cells in the synovial microenvironment. PMID- 15880822 TI - Ultraviolet radiation-induced injury, chemokines, and leukocyte recruitment: An amplification cycle triggering cutaneous lupus erythematosus. AB - OBJECTIVE: To investigate the activation and recruitment pathways of relevant leukocyte subsets during the initiation and amplification of cutaneous lupus erythematosus (LE). METHODS: Quantitative real-time polymerase chain reaction was used to perform a comprehensive analysis of all known chemokines and their receptors in cutaneous LE lesions, and the cellular origin of these chemokines and receptors was determined using immunohistochemistry. Furthermore, cytokine- and ultraviolet (UV) light-mediated activation pathways of relevant chemokines were investigated in vitro and in vivo. RESULTS: In the present study, we identified the CXCR3 ligands CXCL9 (interferon-gamma [IFNgamma]-induced monokine), CXCL10 (IFNgamma-inducible protein 10), and CXCL11 (IFN-inducible T cell alpha chemoattractant) as being the most abundantly expressed chemokine family members in cutaneous LE. Expression of these ligands corresponded with the presence of a marked inflammatory infiltrate consisting of mainly CXCR3 expressing cells, including skin-homing lymphocytes and blood dendritic cell antigen 2-positive plasmacytoid dendritic cells (PDCs). Within cutaneous LE lesions, PDCs accumulated within the dermis and were activated to produce type I IFN, as detected by the expression of the IFNalpha-inducible genes IRF7 and MxA. IFNalpha, in turn, was a potent and rapid inducer of CXCR3 ligands in cellular constituents of the skin. Furthermore, we demonstrated that the inflammatory CXCR3 ligands cooperate with the homeostatic chemokine CXCL12 (stromal cell derived factor 1) during the recruitment of pathogenically relevant leukocyte subsets. Moreover, we showed that UVB irradiation induces the release of CCL27 (cutaneous T cell-attracting chemokine) from epidermal compartments into dermal compartments and up-regulates the expression of a distinct set of chemokines in keratinocytes. CONCLUSION: Taken together, our data suggest an amplification cycle in which UV light-induced injury induces apoptosis, necrosis, and chemokine production. These mechanisms, in turn, mediate the recruitment and activation of autoimmune T cells and IFNalpha-producing PDCs, which subsequently release more effector cytokines, thus amplifying chemokine production and leukocyte recruitment, finally leading to the development of a cutaneous LE phenotype. PMID- 15880825 TI - A novel approach to protein expression profiling using antibody microarrays combined with surface plasmon resonance technology. AB - We have previously described our systems for the high-throughput production of antibodies against mouse KIAA proteins and their validation (Proteomics 2004, 4, 1412-1416). Using our "libraries" of antibodies, we established a novel antibody microarray system in which surface plasmon resonance (SPR) technology is utilized for signal detection. Up to 400 real-time antibody-target bindings could be measured simultaneously within a single hour. This rapid detection was achieved by direct readout of the bindings using SPR technology. To evaluate our system, we assessed the reproducibility on crude protein samples and obtained satisfactorily reproducible results, exhibiting correlation values >0.92. Using this SPR-based antibody microarray system, we examined mKIAA protein expression in five different adult mouse tissues and identified the specific tissue expression patterns of several mKIAA proteins. PMID- 15880826 TI - Art-loving bugs: the resurrection of Spinello Aretino from Pisa's cemetery. AB - During 1391-1392, Spinello Aretino painted a cycle of frescoes in Pisa's cemetery on the theme "Storie dei Santi Martiri Efisio e Potito", highly prized by contemporaries and by Vasari. Twenty years ago, one of these frescoes, "Conversione di S. Efisio e Battaglia" (Conversion of S. Efisio and Battle), because of discolouring and bad damage caused by humidity and atmospheric pollution, was removed from the walls using the "tear-off" technique, consisting in covering the surface with a strong cloth bound to the surface with generous layers of formaldehyde-treated glue. As luck would have it, this large fresco (3.50x7.80 m) was abandoned in a storehouse for more than 20 years. When the curators attempted to remove the cloth, much to their dismay they found that the glue resisted any attempt at digestion, even when treated with concoctions of the most aggressive proteolytic enzymes available on the market. It is likely that during the long storage the glue became slowly cross-linked by the formaldehyde to the point of forming an intricate mass of untreatable proteinaceous material. Thus, although poor Spinello died presumably as a bona fide Christian, his painting was condemned to wear the burka, Muslim-fashion, for the rest of its life. When we recently treated the fresco with a suspension of viable Pseudomonas stutzeri cells, these bacteria, although agnostic, were able to fully digest the hardened glue and restore to life Spinello's glorious painting. We show here how proteomics helped us solve the riddle of how these bacteria acted on the burka obscured fresco. PMID- 15880827 TI - Engineering mammalian cell factories for improved recombinant monoclonal antibody production: lessons from nature? AB - In this review we consider how cell specific recombinant monoclonal antibody (Mab) production by engineered mammalian cells can be improved. Whilst it is generally recognized that Mab production is limited post-transcriptionally at folding and assembly reactions, genetic engineering strategies based on overexpression of individual chaperones or foldases in mammalian cells have not reliably increased cell specific Mab production. Given that recent studies have established that chaperones and foldases themselves exist in a large multiprotein complex, which may coordinate the sequential processing of Mabs, we propose that global expansion of all components of the secretory pathway will likely be necessary to generically improve recombinant Mab production by mammalian cells. In this context, what can be learnt from nature? Important recent studies have delineated some of the main cellular pathways involved in the differentiation of B-cells into nature's own high level Mab producers, plasma cells. This is achieved by a dramatic re-programming of cellular function where the coordinated expansion of metabolic and secretory machinery precedes Ig production, then is maintained by induction of a key intracellular signaling pathway, the unfolded protein response (UPR). Here we review genetic engineering strategies to increase cell specific production rate and discuss whether manipulation of intracellular signaling systems such as the UPR will provide a novel means to engineer mammalian cells for high level recombinant Mab production. PMID- 15880828 TI - The effect of whole cell immobilisation on the biotransformation of benzonitrile and the use of direct electric current for enhanced product removal. AB - The nitrilase of Rhodococcus rhodochrous performs a one-step biotransformation of nitriles to their corresponding carboxylic acids. Application of a direct electric current moves the charged carboxylic acid towards an anode, across an anion exchange membrane, into a separate compartment. Cells encapsulated within alginate beads (2.9 mm diameter) for protection against the current biotransformed benzonitrile to benzoic acid with a 26% reduction in the biotransformation rate, from 0.054 mmol/min/g dcw with free cells to 0.040 mmol/min/g dcw with immobilised cells. When the electric current was applied, the biotransformation rate increased to 0.047 mmol/min/g dcw and product recovery increased from 19% to 79%. PMID- 15880829 TI - Systemic lupus erythematosus in three ethnic groups: XVI. Association of hydroxychloroquine use with reduced risk of damage accrual. AB - OBJECTIVE: To examine whether hydroxychloroquine (HCQ) usage is associated with a reduced risk of damage accrual in patients with systemic lupus erythematosus (SLE). METHODS: Patients (n = 518) meeting the American College of Rheumatology criteria for diagnosis of SLE and with 0.05) between preparation methods, except for group II (high-speed/Ketac Molar) that showed higher infiltration; regarding the materials, Ketac Molar demonstrated the highest microleakage values (p < 0.05), and only Vitremer sealed completely both margins of restorations. It was concluded that air abrasion preparation did not influence microleakage in class V restorations with the employed glass ionomer cements. PMID- 15880931 TI - A study of the relationship between mass and physical strength of keratin bars in vivo. AB - A study was undertaken of the changes in the mass and physical properties of keratin bars implanted subcutaneously in adult rats. A very gradual decrease occurred in vivo in the dry weight of the bars over the period of the study (up to 18 weeks). The elastic modulus of the bars decreased abruptly when present in vivo between 3 and 6 weeks. At the same time there was an increase in the number of cavitations and fissures at the surface of the bars, and an increase in a central internal region of the bars where there was a disorganisation in structure of the polymer. A biocompatible material showing such changes in vivo is likely to be suitable for a variety of medical and surgical applications in which it provides a framework for cell invasion. PMID- 15880932 TI - Influence of thermocycling on the optical properties of laboratory resin composites and an all-ceramic material. AB - The purposes were to determine the difference of color and translucency of dental laboratory resin composites and an all-ceramic material, and to compare the changes in optical properties after thermocycling. Three shades for enamel appearance of two laboratory resin composites (ART; Artglass, TES; Tescera) and all-ceramic material (EMP; Empress 2) were chosen, and three disk specimens for each shade and thickness (0.8, 1.3 and 1.8 mm) were made. Color and spectral reflectance were measured before and after thermocycling over white and black backgrounds. Contrast ratio (CR) and translucency parameter (TP) were calculated. Before and after thermocycling, CR of 1.8 mm specimens did not differ between the materials. After thermocycling, CR and TP of TES were not changed significantly; however, ART and EMP showed increased CR and decreased TP in some shades (p = 0.05). Color change of three materials after thermocycling was not significant. CR increased in inverse proportion to TP (r = - 0.93, p < 0.01). In TES, change of translucency after thermocycling was lower than those of other materials regardless of the specimen thickness. TES seemed to yield more predictable curing performance. PMID- 15880933 TI - Crystallization behavior of silica-calcium phosphate biocomposites: XRD and FTIR studies. AB - Silica and calcium phosphates (CaP) are the most important ingredients in bioactive materials that bond to bone and enhance bone tissue formation. In this study, silica-calcium phosphate (SiO2-CaP) composites were developed by powder metallurgy method, using silica (SiO2) and anhydrous dicalcium phosphate (CaHPO4) powders (CaP) in the ratios (wt%): 20/80, 40/60, 60/40 and 80/20. The effects of temperature and chemical composition on crystallization and phase transformation of the SiO2-CaP composites were evaluated by XRD and FTIR. Thermal treatment of the starting material suggested that CaHPO4 transforms into: gamma-Ca2P2O7 at 800 degrees C; beta-Ca2P2O7 at 1000 degrees C and alpha-Ca2P2O7 at 1200 degrees C. On the other hand, beta-quartz was the only detected phase after thermal treatment of silica in the temperature range 800-1200 degrees C. For all SiO2-CaP composites, SiO2 and CaP did not modify the crystallization behavior of each other when sintered in the temperature range 800-1000 degrees C. However, at 1200 degrees C, CaP promoted the transformation of gamma-quartz into alpha cristobalite. Moreover, SiO2 stabilized beta-Ca2P2O7. The modifications in the crystallization behavior were related to ion substitution and formation of solid solutions. PMID- 15880934 TI - Characterization and bond strength of electrolytic HA/TiO2 double layers for orthopedic applications. AB - Insufficient bonding of juxtaposed bone to an orthopedic/dental implant could be caused by material surface properties that do not support new bone growth. For this reason, fabrication of biomaterials surface properties, which support osteointegration, should be one of the key objectives in the design of the next generation of orthopedic/dental implants. Titanium and titanium alloy have been widely used in several bioimplant applications, but when implanted into the human body, these still contain some disadvantages, such as poor osteointegration (forming a fibrous capsule), wear debris and metal ion release, which often lead to clinical failure. Electrolytic hydroxyapatite/titanium dioxide (HA/TiO2) double layers were successfully deposited on titanium substrates in TiCl4 solution and subsequently in the mixed solution of Ca(NO3)2 and NH4H2PO4, respectively. After annealing at 300 degrees C for 1 h in the air, the coated specimens were evaluated by dynamic cyclic polarization tests, immersion tests, tensile tests, surface morphology observations, XRD analyses and cells culture. The adhesion strength of the HA coating were improved by the intermediate coating of TiO2 from 11.3 to 46.7 MPa. From cell culture and immersion test results, the HA/TiO2 coated specimens promoted not only cells differentiation, but also appeared more bioactive while maintaining non-toxicity. PMID- 15880935 TI - Effect of solid/solution ratio on apatite formation from CaSiO3 ceramics in simulated body fluid. AB - The effect of the solid/solution (S/S) ratio on apatite formation from CaSiO3 ceramics in simulated body fluid (SBF) was investigated. CaSiO3 ceramics with a Ca/Si ratio of 0.91 were prepared by sintering CaSiO3 powder coprecipitated from ethanol solutions of Ca(NO3)2. 4H2O and Si(OC2H5)4 using NH4OH as the precipitant. These ceramics were reacted with SBF at S/S ratios of 1.0, 2.5 and 8.3 mg/ml at 36.5 degrees C for various times. Formation of apatite was observed at all the S/S ratios after soaking for 1 day. The amount and microstructure of the apatite obtained at a S/S ratio of 8.3 mg/ml, however, differed largely from the product formed at the other two S/S ratios. The apatite formed at S/S = 8.3 mg/ml was of smaller particle size, formed in smaller amount and with less preferred orientation of the (001) of apatite crystals compared with those formed at S/S = 1.0 and 2.5 mg/ml. An increase of Ca and decrease of the P components occurred in the soaked SBF at S/S = 8.3 mg/ml, the changes being much more marked than with the other two S/S ratios. These differences in the concentration changes in SBF at different S/S ratios are attributed to the difference in the apatite formation from the CaSiO3 ceramics. PMID- 15880936 TI - Gene expression profiling of bone cells on smooth and rough titanium surfaces. AB - Titanium (Ti) and Ti alloys are widely used as dental and orthopaedic implants, but the effects of the surface characteristics of these materials on the response of cells and target tissues is not well understood. The present study has therefore examined the effects of a rough Ti (RT) and a smooth Ti (ST) surface on human bone cells in vitro. Scanning electron microscopy showed attachment and spreading of cells on both surfaces. Expression profiling using ATLAS gene arrays showed marked differences in gene responses after 3 h of culture. A number of osteoblast genes were identified as "roughness response" genes on the basis of changes in expression on the RT compared with the ST surfaces. The surface roughness of Ti was thus found to have a profound effect on the profile of genes expressed by the bone cells, and suggests that improvements in the biological activity and possibly the clinical efficacy of these materials could be achieved by selective regulation of gene expression mediated by controlled modification of Ti surface. PMID- 15880937 TI - Chromosomal aberrations induced by high-energy iron ions with shielding. AB - Biophysical models are commonly used to evaluate the effectiveness of shielding in reducing the biological damage caused by cosmic radiation in space flights. To improve and validate these codes biophysical experiments are needed. We have measured the induction of chromosomal aberrations in human peripheral blood lymphocytes exposed in vitro to 500 MeV/n iron ion beams (dose range 0.1-1 Gy) after traversing shields of different material (lucite, aluminium, or lead) and thickness (0-11.3 g/cm2). For comparison, cells were exposed to 200 MeV/n iron ions and to X-rays. Chromosomes were prematurely condensed by a phosphatase inhibitor (calyculin A) to avoid cell-cycle selection produced by the exposure to high-LET heavy-ion beams. Aberrations were scored in chromosomes 1, 2, and 4 following fluorescence in situ hybridization. The fraction of aberrant lymphocytes has been evaluated as a function of the dose at the sample position, and of the fluence of primary 56Fe ions hitting the shield. The influence of shield thickness on the action cross-section for the induction of exchange-type aberrations has been analyzed, and the dose average-LET measured as a function of the shield thickness. These preliminary results prove that the effectiveness of heavy ions is modified by shielding, and the biological damage is dependent upon shield thickness and material. PMID- 15880938 TI - Olfactory disorders and their consequences for quality of life. AB - Olfactory disorders are common in the general population, but research into the consequences of these disorders has been lacking. Not until recently, when specific tools for assessing changes in quality of life due to olfactory disorders have become available, have systematic investigations been conducted. In this article we present the most important roles that olfaction plays in humans and review the scientific literature on the consequences of olfactory disorders. This review suggests that quality of life, regarding safety issues and interpersonal relations, as well as eating habits and nutritional intake are severely altered in a large proportion of patients with olfactory disorders. PMID- 15880939 TI - Teratoma of the middle ear: a real entity or a non-entity? AB - Space-occupying lesions of the middle ear range from wholly benign developmental anomalies to highly virulent and aggressive malignancies. Amongst the benign entities classed with the family of developmental anomalies, the middle ear choristoma is well known. But what of the middle ear teratoma? Recent reports suggest that, while rare, teratomas of the middle ear do exist. Middle ear teratomas are defined as tumors consisting of benign tissues derived from all three embryologic layers: endoderm, mesoderm and ectoderm. They are lesions of the neonatal period and early infancy. While complete surgical excision will be curative, a thorough histologic examination of the specimen is recommended to exclude the possibility of immature and/or frankly malignant elements being admixed with the benign teratomatous elements. PMID- 15880940 TI - Merkel cell carcinoma of the auricle. AB - Merkel cell carcinoma (MCC) is an uncommon cutaneous neoplasm which arises in adults with a peak incidence in the sixth and seventh decades. MCC most often arises in the head and neck area and extremities. Among head and neck primary sites, auricular MCC has proven to be rare and only 20 cases have been reported in the literature. Auricular MCC follows the same aggressive course as has been documented for MCC arising elsewhere: the tumor has a propensity for recurring locally and metastasizing to regional lymph nodes and distant sites. Location of MCC in the auricular regions does not appear to confer any survival advantage compared with MCC arising elsewhere. The mainstay of treatment is surgery, with attempts at complete surgical excision being of paramount importance. What role regional node dissection, radiation therapy and chemotherapy may play in the standard treatment of auricular MCC remains to be clearly established. PMID- 15880941 TI - Substance P upregulates osteoclastogenesis by activating nuclear factor kappa B in osteoclast precursors. AB - CONCLUSION: SP upregulates osteoclastogenesis by activating NF-kappaB in osteoclast precursors. OBJECTIVE: Osteoclastic bone resorption is a common finding of otitis media with or without cholesteatoma. Factors upregulating osteoclastogenesis are implicated in bone resorption. The aim of this study was to investigate the effect of substance P (SP) on mouse osteoclastogenesis in vitro. MATERIAL AND METHODS: The SP dose response was analyzed in a mouse osteoclast culture induced by macrophage colony-stimulating factor and receptor activator for nuclear factor kappa B (NF-kappaB) ligand. Western blot analysis was performed to elucidate the subcellular target of SP in osteoclastogenesis. RESULTS: Treatment with SP significantly increased osteoclastogenesis compared to control. SP activated nuclear translocation of NF-kappaB, which is an essential event in osteoclastogenesis in osteoclast precursors (bone marrow macrophages). PMID- 15880942 TI - Sustained extracellular signal-regulated kinase1/2 mitogen-activated protein kinase signalling is related to increased p21 expression in cholesteatoma epithelium. AB - CONCLUSION: These results show for the first time that the RAS/RAF/ERK1/2 MAPK signalling pathway is active and involved in p21-mediated cell cycle arrest in human cholesteatoma epithelium. OBJECTIVE: In a previous report we have demonstrated that the epithelium in human cholesteatoma is characterized by high p53-dependent p21 expression. The RAS/RAF/extracellular signal-regulated kinase (ERK)1/2 mitogen-activated protein kinase (MAPK) signalling pathway can induce p21 expression and subsequent cell cycle arrest via p53-dependent or -independent mechanisms. We designed the present study to investigate whether the RAS/RAF/ERK1/2 MAPK signalling pathway is involved in p53-dependent and p21 mediated cell cycle arrest in human cholesteatoma. MATERIAL AND METHODS: A total of 18 cholesteatoma samples and 18 paired control retro-auricular skin samples were immunohistochemically stained for p53, p21, phosphorylated ERK1/2 (pERK1/2) and total ERK1/2. Positive cells were counted by means of digital image analysis. Double-label fluorescence immunohistochemistry was performed to demonstrate co expression of p21 and pERK1/2. RESULTS: Protein expression of p53, p21 and pERK1/2 differed significantly between cholesteatoma epithelium and retro auricular skin (p <0.01). In cholesteatoma, co-expression of p21 and pERK1/2 was prominent, whereas in retro-auricular skin there was hardly any co-expression. Positive correlations were found between p53 and p21 (p =0.003) and between p21 and pERK1/2 (p =0.013). PMID- 15880943 TI - Effect of click duration on vestibular-evoked myogenic potentials. AB - CONCLUSIONS: The 0.5-VEMP demonstrated a more prominent waveform morphology than either the 0.1- or 0.2-VEMPs. In addition, the 0.5-VEMP had smaller interaural latency differences than the 1.0-VEMP. These findings suggest that 0.5 ms is superior to other click durations in terms of yielding VEMP responses for clinical use. OBJECTIVE: In order to establish the ideal stimulus condition for vestibular-evoked myogenic potentials (VEMPs), we studied the use of various click durations to generate different response patterns in normal subjects. The influence of click durations on VEMPs is described and the optimal stimulation duration for clinical use is suggested. MATERIAL AND METHODS: This was a prospective study. Eighteen healthy volunteers (36 ears) underwent VEMP tests. Four click durations (0.1, 0.2, 0.5 and 1.0 ms) were used in a random order to elicit VEMP responses (0.1-, 0.2-, 0.5- and 1.0-VEMP, respectively). The latency of each peak (p13, n23), the peak-to-peak interval and amplitude (p 13-n23) and the relative amplitude (defined as the amplitude divided by that of the 0.5-VEMP) were measured and compared. RESULTS: Click stimulation of 34 ears (94%) produced 0.1-VEMP responses, whereas positive 0.2-, 0.5- and 1.0-VEMP responses were observed in 36 (100%). The latencies of peaks p13 and n23 were significantly prolonged between successive stimulus durations from 0.1 to 1.0 ms (p <0.05), in contrast to the p13-n23 intervals (p >0.05). The 1.0-VEMP displayed the largest SDs of latencies and interval among the four different VEMPs. The relative amplitude was significantly increased between successive durations from 0.1 to 0.5 ms (alphaT <0.05), but there was no significant difference between 0.5 and 1.0 ms (alphaT >0.05). PMID- 15880944 TI - Dix-Hallpike maneuver results are not influenced by the time of day of the test. AB - CONCLUSION: The result of the DH maneuver does not appear to be affected by the time of day at which it is performed. OBJECTIVE. To determine whether the time of day at which the Dix-Hallpike (DH) maneuver is performed influences the result. MATERIAL AND METHODS: This was a retrospective study. We reviewed the records of all electronystagmagrams (n = 1220) performed at our facility between January 2001 and January 2003, looking at the results of the DH maneuver, the medical history of the patients and the time of day of the test. The distribution of the time of day at which the DH maneuver was performed and induced a positive response was compared to the distribution of the time of day at which all the tests were performed. RESULTS: The distribution of the times during the day when the DH maneuver was positive was not statistically different from that when the DH maneuver was negative. PMID- 15880945 TI - Speech perception results in children implanted with Clarion devices: Hi Resolution and Standard Resolution modes. AB - CONCLUSIONS: Formal testing showed that HiRes users seem to make significant use of acoustic information. Furthermore, from observations reported by experienced care-givers, for example, family, teachers and speech therapists, these children learn a lot from their surrounding environment. Incidental learning, which takes place when acquiring skills or knowledge through naturally occurring events, is a key become available to these deaf paediatric cochlear implant recipients. OBJECTIVE: To compare speech perception skills in children with a Clarion cochlear implant using different speech coding strategies, such as continuous interleaved sampling (CIS), simultaneous analogue stimulation (SAS) and Hi Resolution (HiRes). MATERIAL AND METHODS: The study population comprised 40 children, 17 implanted with a Clarion Hi-Focus 1.2 and 23 with a Clarion CII. All children were pre-lingually deaf and differed in terms of age and cause of deafness. All children had undergone a trial (minimum 6 months) with hearing aids before implantation. Children implanted with a Clarion 1.2 were either CIS or SAS users [Standard Resolution mode (SRM) group]; children implanted with a Clarion CII were Hi-Resolution users [Hi-Resolution mode (HRM) group]. Findings were assessed according to-Erber's hierarchical model (detection, discrimination, identification, recognition and comprehension), making use of a battery of speech perception tests calibrated to the age of the child. Further information concerning use of the implant in everyday situations was obtained by means of the Meaningful Auditory Integration Scale (MAIS) questionnaire, which was administered to the parents. Tests were carried out prior to each fitting session, at switch-on and then at 3, 6, 9 and 12 months. Findings at pre implantation and at 12 months follow-up were collected for both the SRM and HRM groups. Speech perception results were analysed for the SRM and HRM groups, independent of age at implantation, for five subgroups of children according to the paediatric test battery in use and for two subgroups of children, one < and one > 5 years of age. RESULTS: Clarion-implanted children using the Hi-Resolution strategy can develop better speech perception skills at 12 months post implantation compared to children fitted with the SAS or CIS strategy. SAS or CIS users implanted before the age of 5 years tend to achieve better results at 1 year follow-up than children implanted later. In contrast, in Hi-Resolution users, a trend towards better results for recognition and comprehension tasks was observed in children implanted after 5 years of age. PMID- 15880946 TI - Effect of stimulation repetition rate on galvanic-evoked vestibulo-collic reflexes. AB - CONCLUSION: Taking examination time and patient compliance into consideration, we propose that 5 Hz is the optimal stimulation rate of galvanic vestibular-evoked myogenic potentials for clinical use. OBJECTIVES: To evaluate the influence of stimulation repetition rate on galvanic-evoked vestibulo-collic reflexes and to propose the optimal stimulation rate for clinical use. MATERIAL AND METHODS: Both ears of 30 healthy adults were tested at 5 different galvanic stimulation rates (1, 3, 5, 7 and 9 Hz) in a random order. RESULTS: Responses were evident in all 60 ears only at 5 Hz; some ears showed no response at the other frequencies. The relative amplitudes in individual ears were higher at 1, 3 and 5 Hz than at 7 and 9 Hz. Comparison of the latencies of p13g and n23g showed no significant difference among the five stimulation rates. PMID- 15880947 TI - Brainstem auditory-evoked responses in full-term newborn infants with temporary low Apgar score. AB - CONCLUSIONS: No abnormalities, with the exception of maturational changes, in BAER were found during the neonatal period. The results suggest that a temporary low Apgar score is not accompanied by any significant auditory impairment. OBJECTIVE: To examine brainstem auditory function in newborn infants with a temporary low Apgar score but no clinical signs of hypoxic-ischaemic encephalopathy (HIE). MATERIAL AND METHODS: The subjects were 36 full-term infants with Apgar scores of < or =7 at 1 and/or 5 min and > or =8 at 10 min but without HIE. The brainstem auditory-evoked response (BAER) was serially recorded at click rates of 21, 51 and 91/s on Days 1, 3, 5, 7 and 30 after birth. RESULTS: On Day 1 and Days 3-5, the latencies of waves I, III and V tended to increase slightly at all click rates but did not differ significantly from normal control values. Thereafter, all latencies tended to decrease, reaching control values on Day 30. The I-V interval was similar to the control values at all click rates during the first 5 days, tended to decrease from Day 7 and did not differ from the control values on Day 30. There were no significant changes in BAER wave amplitudes at any of the click rates on any day. PMID- 15880948 TI - Histopathologic characteristics of chronic sinusitis with bronchial asthma. AB - CONCLUSION: These findings suggest that patients with both sinusitis and asthma present the histopathologic characteristic of a marked chronic inflammatory reaction, and that eosinophil infiltration may play a significant role in this marked inflammation of the sinus mucosa. OBJECTIVE: Chronic sinusitis and bronchial asthma are known to be closely related. However, the appearance of the mucosa in chronic sinusitis patients with asthma is somewhat different from that in patients without asthma. MATERIAL AND METHODS. We compared the sinus mucosal histopathologies of asthmatic patients with those of non-asthmatic patients. Fifty-three sinusitis patients with a diagnosis of asthma and 54 sinusitis patients without asthma, who served as controls, were enrolled in the study. All of these patients underwent endoscopic sinus surgery. The following seven light microscopic findings were compared in the asthmatic and non-asthmatic groups: the thickness of the basement membrane, goblet cell hyperplasia, subepithelial edema, submucous gland formation, eosinophil infiltration, lymphocyte infiltration and polymorphonuclear leukocyte infiltration. In addition, we explored a possible link between asthma and sinusitis by comparing the following factors in asthmatic and non-asthmatic patients: the presence of allergy, the degree of preoperative polyposis and the extent of preoperative disease as scored by means of ostiomeatal unit CT findings. RESULTS: No statistically significant differences were found between the two groups in terms of the presence of allergy, the degree of preoperative polyposis or the extent of preoperative disease. Basement membrane thickening, goblet cell hyperplasia and eosinophil infiltration were more prominent in the asthmatic compared to the non-asthmatic group (p <0.05). PMID- 15880949 TI - Glycosoaminoglycans in nasal polyps. AB - CONCLUSION: The qualitative and quantitative compositions of GAGs were comparable in all the polyps examined. OBJECTIVE: Glycosoaminoglycans (GAGs) are an integral component of proteoglycans, which are constituents of connective tissue. The qualitative and quantitative compositions of GAGs occurring in proteoglycans determine their biological role. In this work, individual fractions of GAGs occurring in nasal polyps were isolated and estimated. MATERIAL AND METHODS: Polyps were obtained over a 2-year period from 31 patients (18 males, 13 females; age range 28-70 years) who underwent polypectomy and evaluated using routine histopathology. RESULTS: The amount of hyaluronic acid in nasal polyps was high, the amounts of dermatane sulphate and chondroitine-6-sulphate were slightly lower and the amounts of chondroitine-4-sulphate, heparin, heparan sulphate and keratan sulphate were the lowest. PMID- 15880950 TI - Evaluation of mutations in penicillin binding protein-3 gene of non-typeable Haemophilus influenzae isolated from the nasopharynx of children with acute otitis media. AB - CONCLUSION: Younger children tend to harbor more resistant strains because they are exposed to these pathogens more often through contacts with siblings or attendance at day-care centers and are frequently treated with antibiotics. The high prevalence of BLNAR strains should be taken into account in the treatment of AOM in young children. OBJECTIVE: Non-beta-lactamase-producing ampicillin resistant (BLNAR) strains with mutations in penicillin-binding protein (PBP) genes of Haemophilus influenzae have been prevalent recently among younger children. MATERIAL AND METHODS: We investigated mutations in the ftsI gene encoding PBP-3 of H. influenzae isolated from the nasopharynx of children with acute otitis media (AOM) using polymerase chain reaction (PCR). RESULTS: Strains containing the bla gene (beta-lactamase-producing ampicillin-resistant) were identified in 4.7% of cases. Strains with mutations in the ftsI gene (BLNAR) were identified in 23.3% of cases. Strains without mutations in the ftsI gene and that did not contain the bla gene (non-beta-lactamase-producing ampicillin susceptible) were identified in 70.7% of cases. Strains with both expression of the bla gene and mutations in the ftsI gene (beta-lactamase-producing amoxicillin clavulanate-resistant) were identified in 1.3% of cases. The MICs of ampicillin against the strains evaluated in this study were 0.5-2.0 microg/ml. Cefditoren pivoxil had the lowest MIC90 against the strains (0.06 microg/ml). Strains with mutations in the ftsI gene (BLNAR) were broadly identified among young children. PMID- 15880951 TI - Neuroimmunological activation of the afferent laryngeal neural circuit in experimentally induced laryngeal inflammation. AB - CONCLUSIONS: These results show that laryngeal inflammatory reactions may induce the expression of proinflammatory cytokines along the afferent laryngeal circuit and in nuclei associated with the HPA axis. Local laryngeal inflammation may induce functional and physiologic alterations in the laryngeal neural system via neuroimmunologic reactions. OBJECTIVE: Idiopathic laryngeal disorders associated with various neurologic conditions such as spasmodic dysphonia, idiopathic vocal fold paralysis and sudden infant death syndrome are causally related to upper respiratory tract infections, and it can be speculated that these disorders result in neurophysiologic alterations. The goal of this study was to identify the neurophysiologic effect on the central nervous system of local inflammatory alterations in the larynx. MATERIAL AND METHODS: The expression of c-fos and IL 1beta was identified after injecting saline solution, 10 microg of lipopolysaccharide or 100 microg of lipopolysaccharide into the larynx of 12 rats. RESULTS: The inflammatory cytokine IL-1beta was mainly expressed in the inferior olivary nucleus and raphe nucleus, which are associated with the hypothalamic-pituitary-adrenal (HPA) axis. IL-1beta expression was also found in the nuclei of afferent nervous pathways of the superior laryngeal nerve, such as the nucleus tractus solitarius, nucleus ambiguus, lateral reticular nucleus, magnocellular reticular nucleus and paragigantocellular reticular nucleus. PMID- 15880952 TI - Cyclooxygenase-2 regulates the degree of apoptosis by modulating bcl-2 protein in pleomorphic adenoma and mucoepidermoid carcinoma of the parotid gland. AB - CONCLUSION: These results suggest that COX-2 and bcl-2 protein were overexpressed and that apoptosis was reduced in MEC compared to PMA, and that COX-2 may regulate the degree of apoptosis by modulating bcl-2 protein in PMA and MEC. OBJECTIVE: Cyclooxygenase (COX)-2 plays a crucial role in tumorigenesis and overexpression of COX-2 in vitro accompanied by overexpression of bcl-2 protein has been shown to reduce apoptosis. The purpose of this study was to verify that COX-2 regulates the degree of apoptosis by modulating bcl-2 protein in benign and malignant parotid gland tumors. : We examined archival formalin-fixed, paraffin embedded tissue sections of 10 pleomorphic adenomas (PMAs) and 10 mucoepidermoid carcinomas (MECs) by immunostaining with anti-COX-2, anti-bcl-2 and anti-single stranded DNA (ssDNA) antibodies. Labeling indices of the three antibodies were calculated using computer-assisted image analysis. RESULTS: Labeling indices (mean+/-SD) of anti-COX-2 antibody in PMA and MEC were 2.05+/-1.30 and 11.2+/ 2.95, respectively (p < 0.001), those of anti-bcl-2 antibody were 2.00+/-1.28 and 9.68+/-4.05, respectively (p < 0.001) and those of anti-ssDNA antibody were 8.06+/-2.54 and 2.08+/-1.47; respectively (p <0.001). Correlation coefficients between the labeling indices of anti-COX-2 antibody and anti-bcl-2 antibody, anti bcl-2 antibody and anti-ssDNA antibody and anti-COX-2 antibody and anti-ssDNA antibody were 0.88, -0.75 and -0.76, respectively (p <0.001). PMID- 15880953 TI - Carotid artery resection: preoperative temporary occlusion is not always an accurate predictor of collateral blood flow. AB - CONCLUSION: The morbidity predicted by means of preoperative PET studies does not always correlate with the morbidity experienced after permanent carotid artery occlusion. A pre-resection extracranial-intracranial bypass may be necessary to reduce the risk of neurologic morbidity, in particular when carotid artery resection is planned for tumors involving the skull base. OBJECTIVES: Carotid artery resection is generally considered the only curative treatment for patients with advanced head and neck carcinoma involving the carotid artery. PET can be used during temporary occlusion of the internal carotid artery to assess the safety of the procedure. The aims of this paper were to clarify the risk of carotid artery resection and the benefit of extracranial-intracranial bypass. MATERIAL AND METHODS: Twelve patients diagnosed with head and neck cancer adherent to the carotid artery and in proximity to the skull base who had shown good hemispheric collateral blood flow by means of PET underwent carotid artery resection without preoperative bypass. RESULTS: Of the 12 patients who underwent carotid artery resection without reconstruction, 10 suffered no serious neurologic complications; however, 2 suffered cerebral infarctions intraoperatively. PMID- 15880954 TI - Sulindac sulfide-induced apoptosis in sinonasal cancer cells. AB - CONCLUSIONS: These results demonstrate that sulindac sulfide can induce cell death in maxillary cancer cells, and that sulindac sulfide-induced apoptosis is related to the extracellular signal-regulated kinase/p38 MAPK-caspase 3 signaling pathway. OBJECTIVE: Head and neck cancer is the sixth commonest cancer in the human body. Squamous cell carcinoma accounts for most sinonasal cancers. However, little is known regarding the biochemical mechanism(s) of cell death in sinonasal cancers. Recently, human epidemiological and clinical intervention studies have indicated that sulindac, a non-steroidal anti-inflammatory drug, exhibits chemopreventive activity in colorectal cancer. In this study, we aimed to investigate whether sulindac sulfide can induce apoptosis in sinonasal cancer cells and what type of molecular mechanisms induces the death of sinonasal cancer cells. MATERIAL AND METHODS: Sinonasal cancer cells (Asan Medical Center Head and Neck Cancer 5) were treated with various concentrations of sulindac sulfide. The degree of cell death was determined by means of a fluorescence-activated cell scan and the signal transduction pathway for cell death was examined. RESULTS: Human nasal cavity cancer cells treated with sulindac sulfide underwent cell death, and the induction of apoptosis occurred in a dose-dependent manner. Moreover, sulindac sulfide-induced apoptosis was abolished by treatment with the caspase inhibitor Z-VAD-fmk and the mitogen-activated protein kinase (MAPK) inhibitors PD98059 and SB203580. PMID- 15880955 TI - Salivary gland cancer in Finland 1991--96: an evaluation of 237 cases. AB - CONCLUSION: In this material consisting of various salivary gland carcinomas, stage I, male gender and age were the most powerful predictors of patient outcome. OBJECTIVES: To retrieve the records of all salivary gland cancer (SGC) patients diagnosed in Finland between 1991 and 1996 and to evaluate the incidence, histological type and location of SGC, the treatment given and the outcome. MATERIAL AND METHODS: The records for all SGCs (n =286) diagnosed in Finland between 1991 and 1996 and reported to the Finnish Cancer Registry were retrieved. The histological re-evaluation and retrospective study involved 237 SGC patients. RESULTS: The study population consisted of 125 males and 112 females. The mean age was 59 years (males 61 years, females 58 years). Follow-up was at least 5 years. The commonest tumor location was the parotid gland (n = 152; 64%), followed by the minor salivary glands (n =46; 19%), the submandibular gland (n =38; 16%) and the sublingual gland (n = 1; 0.4%). The most frequent histological types of SGC were adenoid cystic carcinoma (n =65; 27%), mucoepidermoid carcinoma (n =45; 19%) and acinic cell carcinoma (n =41; 17%). Surgery, either alone or in combination with other treatment modalities, was used in 209 cases (88%). Radiotherapy was given to 136 patients (57%), 13 of whom (5%) did not undergo surgery. The 5-year overall survival rate was 56.5%, and for stages I-IV it was 78%, 25%, 21% and 23%, respectively (p <0.001; log-rank test). Of the commonest tumor types, the best 5-year relative survival rate was for patients with acinic cell carcinoma (96%), followed by those with mucoepidermoid (79%) and adenoid cystic carcinoma (74%). PMID- 15880956 TI - Inflammatory myofibroblastic tumor in the larynx: clinicopathologic features and histogenesis. AB - Inflammatory myofibroblastic tumor is extremely rare in the larynx and can mimic a malignant process. We present the case of a 62-year-old male who required tracheotomy due to rapidly progressive stridor. Laryngoscopy showed an exophytic, occlusive tumor located in the right true vocal cord. CT showed an expansive mass measuring 2 x2 x1.3 cm3 and occupying the anterior commissure, with glottic progression to the right true vocal cord. The tumor was completely resected with a CO2 laser via a transoral approach. Histologic examination demonstrated extensive ulceration with the presence of granulation tissue. The specimen was mainly composed of spindle cells arranged in a fasciculated pattern with a myxoid background and focal hyalinization. Immunohistochemical studies revealed positivity of spindle cells to vimentin, muscle-specific actin and smooth muscle actin. The patient showed no evidence of disease 24 months after surgery. PMID- 15880957 TI - A case of metastatic colon adenocarcinoma in the larynx. AB - Metastatic carcinomas in the larynx are uncommon, and laryngeal tumors originating from the colon are extremely rare. We report a case of metastatic laryngeal tumor originating from a colon adenocarcinoma in an 81-year-old female. Only a tracheostomy was performed because the patient presented with multiple metastases in other regions. The diagnosis, route of metastasis, treatment and prognosis of metastatic laryngeal tumors are discussed. PMID- 15880958 TI - Investigate the effect of botulinum toxin type A (BTX-A) on nasal symptoms inpatients with allergic rhinitis. PMID- 15880959 TI - Oral aspects of porphyria. AB - Porphyria is a diverse group of diseases in which the biosynthesis of heme is disrupted by either genetic defects or environmental factors. This review gives an overview of the different types of porphyria and describes possible causes, clinical signs, diagnosis and therapy. In addition, the oral manifestations of porphyria and the potential implications of the disease for dental management are discussed. PMID- 15880960 TI - Tooth wear: diet analysis and advice. AB - Diet analysis and advice for patients with tooth wear is potentially the most logical intervention to arrest attrition, erosion and abrasion. It is saliva that protects the teeth against corrosion by the acids which soften enamel and make it susceptible to wear. Thus the lifestyles and diet of patients at risk need to be analysed for sources of acid and reasons for lost salivary protection. Medical conditions which put patients at risk of tooth wear are principally: asthma, bulimia nervosa, caffeine addiction, diabetes mellitus, exercise dehydration, functional depression, gastroesophageal reflux in alcoholism, hypertension and syndromes with salivary hypofunction. The sources of acid are various, but loss of salivary protection is the common theme. In healthy young Australians, soft drinks are the main source of acid, and exercise dehydration the main reason for loss of salivary protection. In the medically compromised, diet acids and gastroesophageal reflux are the sources, but medications are the main reasons for lost salivary protection. Diet advice for patients with tooth wear must: promote a healthy lifestyle and diet strategy that conserves the teeth by natural means of salivary stimulation; and address the specific needs of the patients' oral and medical conditions. Individualised, patient-empowering erosion WATCH strategies; on Water, Acid, Taste, Calcium and Health, are urgently required to combat the emerging epidemic of tooth wear currently being experienced in westernised societies. PMID- 15880961 TI - Clinical management of dental anxiety: what works for whom? AB - This paper aims to provide an overview of the current knowledge regarding the management of adult dentally anxious patients. Furthermore, an attempt is made to formulate a number of preliminary clinical guidelines, based on the available literature. The findings are discussed in the light of the following four problem areas or types of patients, those with: 1) a mild form of fear or anxiety, 2) a phobia of specific dental procedures or situations, 3) interfering psychiatric symptoms and/or 4) a high treatment need. The literature suggests that particularly the implementation of a high level of predictability during treatment, the training of patients in the use of coping skills, and the application of in vivo exposure to anxiety provoking stimuli are the most appropriate options for the management of anxious dental patients and the reduction of their anxiety level. PMID- 15880962 TI - Six-month polyol chewing-gum programme in kindergarten-age children: a feasibility study focusing on mutans streptococci and dental plaque. AB - AIM: To investigate the use of polyol-containing chewing gums in a day-care centre (kindergarten) setting as a means to affect the growth of mutans streptococci and dental plaque. DESIGN: Over a period of six months, 123 five year-old children chewed xylitol (X group), sorbitol (G group), or did not chew gum (C group). Consumption of xylitol, and sorbitol was 4.5 to 5.0 g per day and subjects consumed in five supervised daily chewing episodes four at the day-care centres and one at home. METHODS: Interproximal dental plaque was sampled at baseline and after six months for a laboratory study of mutans streptococci counts. The Quigley & Hein plaque index procedure was used. Interviews and questionnaires elucidated the acceptability of the programme. RESULTS: Parents and kindergarten personnel regarded the programme as an important, additional procedure to promote better oral health. The children regarded the use of chewing gum as a pleasurable experience. Compared with groups G and C, there was a statistically significant reduction of mutans streptococci in the interproximal plaque in the X group. The Quigley & Hein plaque index scores tended to decrease in the X group, while no such trend was observed in the G group. CONCLUSIONS: Habitual use of relatively small daily quantities of polyol-containing chewing gum by young children may be regarded as an important additional caries preventive procedure in a combined day-care centre and home setting. Especially xylitol-containing chewing gum may significantly reduce the growth of mutans streptococci and dental plaque which may be associated with dental caries. PMID- 15880963 TI - Anhydrotic ectodermal dysplasia (Christ-Siemens-Touraine syndrome). A case report. AB - The ectodermal dysplasias (EDs) are a complex group of diseases clinically characterised by congenital absence of ectodermally derived structures. The present report details the features of a 13 year old schoolboy with the rare anhydrotic ectodermal dysplasia (Christ-Siemens-Touraine syndrome). PMID- 15880964 TI - Fluoride concentration of bottled water, tap water, and fluoridated salt from two communities in Mexico. AB - AIM: To determine fluoride levels in bottled water, tap water, and fluoridated salt from two communities in Mexico. DESIGN: Stratified random collection of water and salt samples from Mexico City and Veracruz, Mexico for fluoride analysis. METHODS: Samples were analysed using a combination fluoride ion specific electrode. Results were compared using Student's t-test and mixed-model ANOVA. Water fluoride values were compared by type, community and collection area; salt fluoride values were compared by community and collection area. RESULTS: 197 tap water samples, 133 bottled water samples and 20 fluoridated salt samples were collected. The mean (+/- SD) fluoride content for all tap water was 0.20 +/- 0.17 microg F/g (ranging from 0.01 to 0.88 microg F/g) and 0.24 +/- 0.24 microg F/g for all bottled water (ranging from 0.01 to 2.80 microg F/g). This difference was not statistically significant. When results were analysed by city, the difference between tap water samples was statistically significant. Ten bottled water samples contained more than negligible fluoride (ranging from 0.7 2.8 microg F/g). Mean salt fluoride content was 230.0 +/- 49.8 microg F/g, which was within governmental regulation levels. CONCLUSION: Some water samples had amounts of fluoride exceeding the maximum recommended levels. Salt fluoride levels were within regulation limits. Monitoring of fluoride content of both bottled and tap water is strongly advised. PMID- 15880965 TI - Decreasing caries prevalence in Japanese preschool children is accompanied with a reduction in mutans streptococci infection. AB - OBJECTIVE: To evaluate the association between decreasing caries prevalence in Japanese preschool children and mutans streptococci (MS) infection. METHODS: The prevalence of caries and salivary MS was examined in 1- to 3-year-old preschool children in 1995 (n = 135) and 2000 (n = 118). RESULTS: The caries prevalence for 3-year-old subjects significantly declined from 65.1% to 37.5% (p < 0.01), reflecting the data seen in Japanese national oral health surveys in 1993 and 1999. Salivary MS detection for 3-year-old subjects significantly declined from 68.3% to 37.5% (p < 0.05). On the other hand, the decrease in the prevalence of caries and salivary MS was not significant in 1- and 2-year-olds. The correlation between caries experience (dft) and the salivary MS score for 3-year-old subjects with low caries prevalence in the later examination (r(s) = 0.479, p < 0.001) was comparable with that in 1995 (r(s) = 0.462, p = 0.001). CONCLUSIONS: The recent decrease in decay in Japan is accompanied with a reduction in MS infection in the deciduous dentition. MS levels are closely related to caries prevalence in preschool children regardless of the extent of caries. PMID- 15880966 TI - 21st century endodontics. Part 1. AB - Root canal treatment techniques probably develop and change more frequently than any other area of dental practice and it can be hard for the busy general dental practitioner to keep up to date. The aim of this series of five papers is to give a thorough review of current teaching and practice. This will encompass the rationale of root canal treatment, guidance on the latest technical procedures for preparation and obturation of the entire root canal system, and consideration of how these principles may contribute to successful treatment. The series will address the following issues: diagnosis and treatment planning; root canal morphology and access; control of infection by effective isolation and anti microbial agents; the philosophy of modern canal preparation techniques; single- or multi-visit treatment with relevant canal medication; obturation; diagnosis of failure and considerations in re-treatment. It is hoped that readers will be encouraged by the series to question their own techniques, audit their outcomes, and adopt new techniques only after careful reflection of the underlying principles involved. PMID- 15880967 TI - Dentist-patient relationship and quality care 1. Introduction. AB - Although quality is a genuine concern for dentistry, nowadays more emphasis is placed on quality issues. As dentist-patient interaction is involved in many aspects of care and it is more crucial for dentistry when compared to many other professions, a good dentist-patient relationship is an integral element of quality care. This series of 'practice articles' examines various important dimensions of this interaction. The first and second papers examine the value of trust and communication, the third paper focuses on informed consent and the fourth paper evaluates the relatively broadened role of dentists in behavioural modification. PMID- 15880968 TI - Global maternal and child health. PMID- 15880969 TI - Risk of low birth weight associated with advanced maternal age among four ethnic groups in the United States. AB - OBJECTIVES: To examine and compare the risk of low birth weight associated with delayed childbearing in four ethnic groups using nationally representative data in the United States. METHODS: We compared the risk of low (< 2.5 kg) birth weight among African Americans, Mexican Americans, Puerto Ricans, and non Hispanic whites using birth data for the United States obtained from the National Center for Health Statistics. Comparisons were done separately for first births and births of second or higher order and in terms of odds ratios, risk differences and attributable fractions of very low (< 1.5 kg), middle low (1.5 2.5) and overall low birth weight. Statistical analysis included use of logistic regression models with likelihood ratio tests for interaction effects. RESULTS: African Americans and Puerto Ricans, and to a lesser extent Mexican Americans, had higher risk differences associated with advanced maternal age. For first births, the risk differences associated with advanced maternal age (> or = 35 years) in low birth weight were 5.3% (95% CI, 4.7-6.0), 4.3% (95% CI, 1.7-6.9), and 3.7% (95% CI, 2.8-4.5) for African Americans, Puerto Ricans, and Mexican Americans, respectively, as compared with 2.6% (95% CI, 2.4-2.7) for non-Hispanic whites. On the other hand, the odds ratios associated with advanced maternal age were more similar across the four ethnic groups. Differences were greater for all ethnic groups in the case of first births as compared with births of second or higher order. CONCLUSIONS: Advanced maternal age appears to be associated with for the most part similarly increased odds of low birth weight for African Americans, Mexican Americans, Puerto Ricans, and non-Hispanic whites. However, the age-related increments in the risk of low birth associated with advanced maternal age are greater for African Americans, Puerto Ricans and, to a lesser extent, Mexican Americans, as compared with non-Hispanic whites. PMID- 15880970 TI - Association of acculturation with cesarean section among Latinas. AB - OBJECTIVES: Examine the association of acculturation and cesarean section after adjusting for clinical and non-clinical factors that could influence clinical discretion in performing the surgery. METHODS: A sample of 2102 low-risk, low income primarily Mexican Latinas in San Diego County was divided into two groups: primiparas and multiparas. For each parity group, logistic regression was used to assess the association of acculturation and cesarean section. RESULTS: Among multiparous Latinas, the risk of cesarean section for highly acculturated women exceeded the risk for the less-acculturated women, but the result was reverse for primiparous women. The adjusted relative odds of cesarean section were twice as high [OR 2.1, 95%CI 1.1-4.1] for multiparous US-born Latinas relative to multiparous Spanish-speaking women born in Mexico. While for primiparous women this same comparison showed US-born Latinas to be approximately half as likely to have a cesarean delivery [OR 0.4, 95%CI 0.2-0.7]. CONCLUSIONS: In order to reduce the chances of unnecessary cesarean sections among Latinas, the role of acculturation in women who have and have not already given birth needs to be investigated further. PMID- 15880971 TI - Prevalence of postpartum depression in a native American population. AB - OBJECTIVES: Data were collected on postpartum depression from 151 women, ages 16 40 years who received postpartum health services from a rural obstetrical clinic in North Carolina between September 2002 and May 2003. Reflective of the racial and socio-economic makeup of the county, 60.9% of the sample were American Indian (Lumbee tribe) 25.8% were African American and 13.3% were Caucasian or other. METHODS: The Postpartum Depression Screening Scale (PDSS) was utilized to explore the prevalence of postpartum depression requiring clinical intervention in a largely unexplored population, minority women. RESULTS: The incidence of postpartum depression symptoms was over 23%, which is significantly higher than even the most liberal estimates in other populations. As with previous literature on risk factors, the sample demonstrates a strong association between symptoms of depression, history of depression and receiving treatment for depression. CONCLUSIONS: The PDSS proved to be a clinically useful tool in this setting. Findings support the importance of implementing routine screening protocols to guide practice and implement support services. PMID- 15880972 TI - Parental awareness of health and community resources among immigrant families. AB - OBJECTIVES: To examine the association between parental immigrant status and awareness of health and community resources to help address common family problems. METHODS: Using the 1999 National Survey of America's Families, a survey of the health, economic, and social characteristics of children and adults, bivariate and multivariate analyses were conducted on 35,938 children to examine the relationship between parents' immigrant status (U.S.-born citizens, naturalized citizens, and noncitizens) and their responses to questions about their awareness of specific health and community resources. RESULTS: Compared to U.S.-born citizens, noncitizens were at the highest risk of not being aware of health and community resources for most outcomes, followed by naturalized citizens. The services of which noncitizens were most likely to be unaware were places to get help for family discord, child care issues, and family violence. Multivariate analyses indicate that parental race/ethnicity, education level, employment status, and child age were other significant independent risk factors. CONCLUSIONS: Immigrant parents are at particularly high risk of alienation from systems of health care and support services that are available to low-income and other vulnerable populations in the United States. These findings clearly document disparate awareness among parents of different immigrant status. Community and health resources should reach out to immigrant populations, in linguistically and culturally appropriate ways, to alert them to the availability of their services. PMID- 15880973 TI - Use of home visit and developmental clinic services by high risk Mexican-American and white non-Hispanic infants. AB - OBJECTIVE: To investigate whether US-born infants of mothers of Mexican descent who were enrolled in Arizona's Newborn Intensive Care Program (NICP) received follow-up services (developmental clinic and community health nurse [(CHN)] home visits) at the rates similar to White non-Hispanic (WNH) infants. Socio-economic and health status characteristics were controlled using stepped regressions in order to assess the impact of each on service use. METHODS: This population-based study used retrospective data from the NICP administrative database that were linked to birth certificates for years 1994-1998. The study population was limited to Arizona-born infants; it included 7442 infants of WNH mothers, 2612 infants of US-born Mexican American (MA) mothers and 2872 infants of Mexico-born mothers. Four service use indicators were used in the analysis. RESULTS: Both Hispanic infant subgroups were less likely to have a CHN visit by 6 months and by 1 year, and to average fewer CHN visits. A smaller percent attended the developmental clinic by age one. After controlling for language, demographics, health status, socio-economic characteristics, and mothers' prenatal care use, infants of US-born MA mothers had rates of use similar to WNH. However, even after controlling for the study variables, infants of Mexico-born mothers were less likely (OR = .83) to use the developmental clinic. Hispanics continued to lag behind in the use of services compared with WNHs. The disparity is not a function of ethnicity, but appears attributable to demographic and socio-economic characteristics. Infants who had a CHN visit were significantly more likely (OR = 2.51) to use the developmental clinic than those without a nurse visit. Infants whose mothers had inadequate prenatal care were less likely to use these follow up services even after controlling for study variables. CONCLUSIONS: Infants whose mothers had inadequate prenatal care should be targeted for more intense CHN visits. Infants of mothers born in Mexico may need additional support/assistance in using the developmental clinic. PMID- 15880974 TI - Measuring primary care for children of Latino farmworkers: reliability and validity of the parent's perceptions of primary care measure (P3C). AB - This study evaluates the feasibility, reliability, and validity of the Parent's Perceptions of Primary Care measure (P3C) in an underserved population: children of Latino farm workers. Bilingual research assistants verbally administered the P3C, as well as a measure of child health-related quality of life (HRQL: the PedsQL 4.0) and demographic questions to 297 Latino farm worker parents of young children, in San Diego and Imperial Counties. The P3C was found to be feasible, as measured by a very low percent of missing/do not know values. Internal consistency reliability for the Total Scale and most subscales was strong. The P3C's validity was demonstrated through factor analysis of the subscales, by showing that scores were lower for children without a regular physician and for children experiencing foregone health care, and by demonstrating that P3C scores were related to HRQL. The P3C can be useful to various stakeholders in measuring primary care for vulnerable populations. PMID- 15880975 TI - Measures of strength for maternal health programs in 55 developing countries: the MNPI study. AB - OBJECTIVES: To measure levels and types of effort for national maternal and neonatal health programs in 55 developing countries, in 2002, as a replication of a 1999 study. METHODS: Thirteen components of program effort were covered, based on 81 items in questionnaires completed by 10-25 expert respondents in each country. RESULTS: With 100% representing maximum effort, the international average was 58-60%, and the 13 component averages varied from 48 to 72%. The components included health center and district hospital capacities, services provided, proportion of the rural and urban populations with actual access to the services, together with the support functions of policy, training, education, resources, and evaluation. Scores are high for policies but low for access, resources, training, and public education. CONCLUSIONS: National programs to improve maternal health are far from satisfactory, as assessed here, with negligible improvement from 1999-2002. Efforts fall short in general, but considerably more so for some program features than others. Literal access to basic services is poor, and is especially lacking in rural areas. Regions differ much more in the access they provide to services than in other respects. PMID- 15880976 TI - The trend of maternal mortality in Egypt from 1992-2000: an emphasis on regional differences. AB - OBJECTIVE: To review factors contributing to a 52% drop in Egypt's maternal mortality ratio (MMR) per 100,000 live births from 174 in 1992-93 to 84 in 2000. METHODS: Data on maternal mortality were collected from all 27 governorates in Egypt between 1 January and 31 December 2000. This round of maternal mortality data is compared with the earlier nation wide maternal mortality study in 1992. Health care interventions that may account for the decrease were reviewed. RESULTS: MMR decreased by 51.7% nation wide. This decrease was greater in the less-developed parts of Upper Egypt (59%), than in Lower Egypt (30%). A multifaceted set of interventions were concentrated in Upper Egypt. The greatest decrease in maternal mortality was associated with the area of highest intervention, greatest need, and during the time period of the implementation of this program. There were increases in use of health services; use of modern contraceptives; hospital deliveries; and use of trained birth attendants. For most indicators, the changes were greater in Upper than Lower Egypt. CONCLUSION: Since 1992-93, efforts by the Government of Egypt and donors to improve access to and the quality and utilization of services can be linked to a greatly reduced MMR. PMID- 15880977 TI - Preventive healthcare of infants in a region of Lebanon: parental beliefs, attitudes and behaviors. AB - OBJECTIVES: This study aimed to investigate healthcare seeking behaviors of mothers for themselves and their infants in the Bekaa Valley, Lebanon, to identify attitudes and beliefs towards the preventive healthcare of infants and to assess whether a healthcare-based intervention program would be accepted by the population. METHODS: Ninety-two interviews were conducted with women who had delivered a live birth in two hospitals in the Bekaa. The hospitals selected attracted women from different socioeconomic classes. RESULTS: Differences were found in adequacy of healthcare utilization based on household size, number of children and prenatal care utilization. There was a higher perception of barriers to healthcare by parents who did not seek adequate preventive care for their infants. Mothers of infants who had inadequate care had a lower perception of severity of illness and of physicians' ability to prevent illness. The population believes strongly that medical professionals are reliable sources of health information and guidance. Infants' mothers and grandmothers are important sources of healthcare-related information. CONCLUSIONS: Rates of preventive infant healthcare in the Bekaa need improvement. Any intervention must target mothers and grandmothers, as they appear to influence the medical care of infants. Other targets include parents of larger families. Dissemination of information regarding the seriousness of illness and the ability to prevent illness is required. The medical profession may influence health choices made by this population. However, further investigation of healthcare barriers, especially economic and geographic barriers, is needed. In preparing to improve healthcare utilization for infants in the Bekaa, local, regional and international organizations should refer to the population's beliefs, attitudes and behaviors described in this study, as these findings may aid in planning interventions that are likely to positively impact mothers and infants. PMID- 15880978 TI - The clustering of risk behaviors among Caribbean youth. AB - OBJECTIVE: To examine the relationships among risk behaviors for Caribbean youth; and to determine the correlations between initiation of sexual activity and other risk behaviors. METHODS: The associations between cigarette smoking, alcohol and marijuana use, early initiation of sexual intercourse, involvement in violence and delinquency were examined using odds ratios on data from the Caribbean Youth Health Survey (n = 15, 695). Survival analysis was then used to determine the association between initiation of sexual activity and the risk behaviors. FINDINGS: There were statistically significant relationships between all pairs of risk behaviors for both male and female adolescents. Even though more males than females had engaged in each of the behaviors, the strengths of association were higher for females. From survival analysis, initiation of sexual activity was associated with gang involvement and weapon carrying among young adolescents and even more risk behaviors among the older adolescents. CONCLUSION: Health compromising behaviors cluster among Caribbean youth with associations being stronger for females. Initiating sexual activity was a predictor of other risk behaviors with the likelihood increasing among older adolescents and females. PMID- 15880979 TI - The relationship between birth predictedness and violence during pregnancy among women in La Paz And El Alto, Bolivia. AB - OBJECTIVES: The main objectives were to estimate the prevalence of predicted and unpredicted last births using a prospective approach and to estimate the prevalence of violence during the last pregnancy. In addition, the relationship between birth predictedness and violence during pregnancy was examined. METHODS: The target population for this study was women who had participated in the 1994 Demographic and Health Survey (DHS) and lived in El Alto and La Paz Bolivia (n = 1308). In 1997, 816 women were located and re-interviewed. During this three-year interval, 127/816 women had given birth to their last child. RESULTS: Of the last births that occurred during the three-year interval, 82% were unpredicted (18% were to women who stated in 1994 that they wanted to postpone childbirth for more than three years and 64% were to women who stated they wanted to wanted to forego childbearing entirely). Twenty-eight percent of women reported that they had experienced violence during their last pregnancy. No statistically significant relationship was found between birth predictedness and violence during their pregnancy. CONCLUSIONS: The majority of births that occurred in the three-year study interval were unpredicted. The prevalence of violence during pregnancy was alarmingly high among this sample of women. Further investigation on violence during pregnancy is needed and should be expanded to examine how violence during pregnancy impacts maternal and infant outcomes, which have remained poor in this country. In addition, the high rates of unpredicted births illustrate that work remains to be done in addressing women's ability to control their fertility. PMID- 15880980 TI - A guide to reviewing manuscripts. AB - MCH professionals play an invaluable role by serving as peer-reviewers for their professional journal. The peer-review process is a fundamental part of maintaining the scientific rigor and professional relevance of our field's literature. The purpose of this article is to provide guidance for the review of manuscripts submitted for publication to professional and scientific journals. In addition, a structure is proposed for undertaking a review and organizing the written response to the editor and the authors. PMID- 15880981 TI - Brugada syndrome--an update. AB - A diagnostic triad characterizes Brugada syndrome. It consists of a right bundle branch block, ST-segment elevation in leads V1-V3 and sudden cardiac death (SCD). Approximately 50% of patients with Brugada syndrome noted to have familial occurrence, this suggests a genetic component of the disease. Mutations in gene SCN5A, an encoder for human cardiac sodium channel on chromosome 3p21, causes Brugada syndrome. Before considering the diagnosis of Brugada syndrome, exclude precordial ST-segment elevation secondary to acute coronary syndrome, electrolyte imbalance, myocarditis, drug over dosage (cocaine, tricyclic antidepressants), and arrhythmogenic right ventricular cardiomyopathy/dysplasia. Intravenous administration of ajmaline, flecainide, and procainamide may exaggerate the ST segment elevation, or unmask it when it is initially absent in patients with suspected Brugada syndrome. Programmed electrical stimulation (PES) may help in risk stratification, and in some cases, establish the diagnosis. However, the accuracy of PES in predicting outcome is debatable, especially in patients showing an asymptomatic Brugada ECG, and reporting no family history of SCD. Treatment with an implantable cardioverter-defibrillator (ICD) is the only established effective therapy for the disease. With ICD therapy, the mortality rate at a 10 year follow-up was 0%. Supporting data for long-term pharmacological therapy with quinidine, or isoproterenol for prevention of SCD, in these patients, is uncomplete. Future advances in understanding the molecular mechanisms of Brugada syndrome may provide answers to many of the controversial issues in the management of this disease. PMID- 15880982 TI - Paradoxical increase in 3-nitropropionic acid neurotoxicity by alpha-phenyl-tert butyl-nitrone, a spin-trapping agent. AB - BACKGROUND: 3-Nitropropionic acid (3-NP), a mitochondrial toxin, impairs cellular energy generation by inhibiting succinate dehydrogenase. The basis of its neurotoxicity is oxidative stress in the wake of cellular energy failure. alpha Phenyl-tert-butyl-nitrone (PBN), a spin-trapping agent with free radical scavenging capability, has shown protective effects in various models of experimental brain insults. The effect of PBN on the 3-NP neurotoxicity paradigm was evaluated in this study. METHODS: Two groups of adult male mice receiving daily systemic 3-NP administration were pretreated with PBN or normal saline respectively for 5 days. After the treatment course, motor dysfunction and the volume of cerebral lesions were quantitatively evaluated. Cellular apoptosis and expressions of glial fibrillary acidic protein (GFAP) and cyclooxygenase-2 (COX 2) in the brain were compared between the 2 groups. RESULTS: All mice treated with normal saline and 3-NP survived but developed mild motor dysfunction. Apoptosis of striatal cells was noted in the absence of destructive cerebral lesions. In contrast, combined treatment with PBN and 3-NP resulted in more severe motor dysfunction and higher mortality in experimental animals. Destructive lesions with cellular necrosis, and enhanced expressions of GFAP and COX-2 were noted in the striatum. CONCLUSIONS: 3-NP neurotoxicity was paradoxically accentuated by the combined treatment with PBN and 3-NP. Metabolic clearance of 3-NP is probably impaired by PBN and the increased oxidative stress caused by higher 3-NP levels may exceed the free radical-scavenging ability of PBN. The shift from apoptotic to necrotic changes with increased 3-NP toxicity is in accord with the theory that cellular energy reserves determine the pattern of cellular death. PMID- 15880983 TI - Ocular complications after a sub-tenon injection of triamcinolone acetonide for uveitis. AB - BACKGROUND: The purpose of this study was to evaluate the complications of a sub Tenon injection of triamcinolone acetonide in patients with uveitis. METHODS: We retrospectively reviewed the records of uveitis cases treated from 1999 to 2003 in Kaohsiung Chang Gung Memorial Hospital. Thirty patients (45 eyes), who received a sub-Tenon injection of 0.5 ml of a triamcinolone acetonide suspension for uveitis control, were included, and 16 (26 eyes) of these patients underwent intraocular pressure (IOP) monitoring for over 3 months after the procedure. RESULTS: The incidence of ocular penetration was 1.6% and of rapidly progressive cataracts was 13.3%. The incidence of ocular hypertension (>21 mmHg) was 76.9% (20/26). The ocular hypertension in most cases was controlled with topical medications, with recovery of normal IOP. Long-term topical antiglaucoma medication was given for 3 eyes, and another 3 eyes (11.5%) received a trabeculectomy performed by glaucoma specialists. The incidence of glaucoma during the follow-up period after treatment is 23.1% (6/26). CONCLUSIONS: The 23.1% incidence of glaucoma in this study is higher than expected. Careful pretreatment evaluation and post-treatment monitoring of IOP are mandatory for patients with uveitis receiving a sub-Tenon injection of triamcinolone acetonide. PMID- 15880984 TI - Tongue-lip adhesion in the management of Pierre Robin sequence with airway obstruction: technique and outcome. AB - BACKGROUND: Airway obstruction and feeding difficulty can occur in patients born with Pierre Robin sequence. In select patients with pronounced micrognathia, surgical intervention to relieve the airway obstruction is necessary. The surgical indications and appropriate surgical procedure continue to have a great deal of controversy. The purpose of this study was to evaluate our experience of tongue-lip adhesion in the management of upper airway obstruction associated with Pierre Robin sequence. METHODS: From March 1995 through May 2002, a total of 14 patients with Pierre Robin sequence, who were admitted to the pediatric neonatal intensive care unit either with prolong intubation, poor body weight gain, or repeated airway infection, underwent tongue-lip adhesion (TLA). The operation was performed by raising mucosa flaps and approximated the muscles between tongue and lower lip. Retention sutures were used. The patients were evaluated for clinical responses. RESULTS: Our successful rate with tongue-lip adhesion was 70%. Ten of the 14 patients showed clinical improvements including extubation of the endotraccheal tube, body weight gain, return for home care, reduced episodes of respiratory infection, and improvement in O2 saturation and blood gas. The surgical procedure was simple to perform without major complications. CONCLUSIONS: This retrospective review showed that with a thorough preoperative airway evaluation, TLA could be successfully used to treat select patients with Pierre Robin sequence suffering from severe upper airway obstruction. Thus, TLA should be first considered when surgical relief of airway obstruction is indicated and when obstruction is limited to the classic tongue base obstruction type. PMID- 15880985 TI - Midpalmar accurate incision for carpal tunnel release. AB - BACKGROUND: Carpal tunnel syndrome is the most common entrapment neuropathy in humans today. For patients in whom conservative treatment fails, surgical decompression is indicated. Among the various surgical techniques currently in use, endoscopic techniques are becoming increasingly popular. Due to the rapid postoperative recovery shown after endoscopic operations, midpalmar accurate incision for carpal tunnel release is a comparative alternative. METHODS: From February 1998 through January 2003, 84 patients had undergone 96 midpalmar carpal tunnel releases with accurate skin incisions under regional block or general mask anesthesia. The postoperative evaluations were performed via subjective assessment with a standardized telephone interview over an average follow-up period of 22.4 months. RESULTS: The average operation time was 16 minutes (range, 5 to 40 minutes). A total of 87 hands (91%) had excellent or good recovery in terms of symptom relief (improvement of 70% or greater). The incidence of postoperative sensory morbidity, i.e., pillar pain or scar tenderness, was 7% (7 hands). The motor functional morbidity of a persistent subjective decrease in grip strength was noted in 8% (8 hands), and the mean period for returning to work was 4.5 weeks. In addition, no operation-induced neurovascular or tendinous injury occurred in any patient. CONCLUSIONS: The outcomes were similar to those of endoscopic carpal tunnel release. Based on these results, midpalmar carpal tunnel release with accurate location of skin incision is as effective as any other surgical procedures for carpal tunnel release. Furthermore, it is also a safe and simple procedure. PMID- 15880986 TI - Use of the Arndt wire-guided endobronchial blocker to facilitate one-lung ventilation for pediatric empyema during video-assisted thoracoscopy. AB - BACKGROUND: Video-assisted thoracoscopic surgery (VATS) has emerged as an innovative and popular procedure for the management of postpneumonic empyema in children refractory to a medical response. One-lung ventilation is required during VATS. In this study, we evaluated the efficacy of intraoperative wire guided endobronchial blockade (WEB) for achieving 1-lung ventilation during a thoracoscopic procedure for pediatric empyema. METHODS: Eighteen patients undergoing a VATS approach for evacuation of an empyema cavity were studied. We used a new device, a bronchial blocker tube, to establish 1-lung ventilation. Intraoperative oxygenation, ventilation, and hemodynamics, as well as the duration of the operation during 1-lung ventilation were recorded. The number of unsuccessful placement attempts, number of malpositionings of the device, and the number of secondary dislodgements of the device after turning the patient into the lateral position were also counted. The quality of lung deflation and inflation was rated by the surgeon under direct visualization as either excellent, fair, or poor. RESULTS: The mean operative time was 80+/-10.8 (range, 50 approximately 120) min. The mean peak inspiratory pressure under 1-lung ventilation was 28.7+/-3.6 cm H2O, and no desaturation was noted. A number of unsuccessful placement attempts were required in 1 patient (1/18) for left-sided VATS. No malpositioning or secondary dislodgement of the device was noted. The quality of lung deflation was judged as being excellent in all patients. CONCLUSIONS: VATS can safely and effectively be performed in children with a proper anesthetic technique. With the development and clinical use of this new device, the bronchial blocker tube proved to be effective and easy to use for establishing 1-lung ventilation in a pediatric population. PMID- 15880987 TI - Lipofibromatous hamartoma of the median nerve with long-term follow-up. AB - Lipofibromatous hamartoma is a rare, benign tumor that most often involves the median nerve. A 16-year-old male with lipofibromatous hamartoma of the median nerve at the wrist level is described. This patient was a child when the mass was first noted. Although there were no symptoms or signs of carpal tunnel compression, the growth of the tumor was progressing. In addition to the release of the carpal tunnel, microsurgical intraneural dissection was done to preserve the thenar motor branch. Then segmental excision of the residual sensory component with sural nerve grafting was performed. Subjectively the patient did not notice the minor motor deficit, however, the patient did experience numbness of fingertips after surgery. There were no scars or trophic ulcers on fingertips at 3 years of follow-up regardless of the inadequate sensory return. Treatment of this benign tumor is still controversial. The relevant reports in the literature are reviewed. PMID- 15880988 TI - Congenital radioulnar synostosis treated using a microvascular free fasio-fat flap. AB - Congenital radioulnar synostosis, which is caused by fusion between the proximal end of the radius and ulna, is an uncommon deformity of the upper extremity. It is characterized by the forearm being fixed in some degree of pronation and patients present with a variety of functional limitations of the involved limb. This condition may lead to difficulties in daily activities, such as writing, eating, and accepting objects in an open palm. Many operative procedures have been developed to mobilize the fixed forearm but the long-term results are disappointing. Herein, we reported a 12-year-old female patient with congenital radioulnar synostosis of the bilateral forearm who was treated with separation of the bony bridge between the left radius and ulna combined with a free groin flap transfer into the separated space. The functional results are good and no recurrent ankylosis occurred during a period of 28 months postoperatively. The flap size and volume of the subcutaneous fat are the key factors for satisfactory results. PMID- 15880989 TI - Intrapartum spontaneous uterine rupture following uncomplicated resectoscopic treatment of Asherman's syndrome. AB - Since Asherman first published his series of intrauterine synechiae in 1948, only a few physicians have described the obstetric complications of patients who conceived following surgical treatment of intrauterine synechiae. We present a woman with a history of resectoscopic resection of intrauterine adhesions with a term pregnancy and spontaneous uterine rupture that occurred during the intrapartum period. At emergent cesarean section, hemoperitoneum of approximately 1500 mL was noted and a 10-cm defect was present in the lateral uterine wall; the edges of the defect were bleeding actively. Because of the potential for a disastrous outcome in the rupture of the pregnant uterus, patients treated for Asherman's syndrome should be identified early and appropriate precautions should be taken in their obstetric management. PMID- 15880990 TI - Ancylostoma duodenale infection: a study of serum immunoglobulin G4 response to the excretory secretory antigen of adult worm. AB - The serum anti-Ancylostoma duodenale immunoglobulin (Ig) G4 antibody response to fraction III of the partially purified excretory secretory antigen of adult worm (Ad III ESA) was studied. The work included 60 patients with A. duodenale infection (GI), 40 patients with other parasitic infections (GII) and 30 apparently healthy parasite-free controls (GIII). Level of serum specific IgG4 was measured by indirect enzyme linked immunosorbent assay and compared with serum specific IgG, IgG 1, 2 & 3 subclass antibodies. Patients of GI had gastro intestinal manifestations and symptoms suggestive of anaemia, and by investigations they had anaemia in 31.7% & eosinophilia in 100%. Measuring the intensity of A. duodenale infection, quantified as fecal egg counts, in patients of GI revealed that 60%, 30% & 10% had light, moderate, and heavy infections, respectively. The serum anti-Ad III ESA IgG & IgG 1-4 subclass antibodies were significantly elevated (P < 0.001) in patients of GI compared with GIII. Serum specific IgG4 was expressed in 100% of patients of GI at a significantly highly elevated level than IgGI (P < 0.01), IgG2 & IgG3 (P < 0.001). Specific IgG1 was expressed in 88.3% of patients of GI at a significantly elevated level (P < 0.001) than IgG2 & IgG3 which were expressed in 31.7% & 38.3%, respectively and elevated to a moderate extent. Serum specific IgG4 showed a 1.0, 1.1, 3.1 & 2.6 fold increase in detection rate of positive cases than IgG, IgG1, IgG2 & IgG3, respectively. The highest ability to differentiate between infected and healthy subjects was by serum specific IgG4 recording a discrimination coefficient of 9.4, while IgG, IgG1, IgG2 & IgG3 recorded 5.2, 6.3, 3.2 & 3.4, respectively. Serum specific IgG4 showed a significant positive correlation (r = 0.41, P < 0.001) with the intensity of A. duodenale infection, as was demonstrated by IgG & IgG3 (P < 0.01 & P < 0.05, respectively). Detection of serum anti-Ad III ESA IgG4 antibody recorded a 100% sensitivity that was significantly higher (P < 0.001) than IgG1, IgG2 & IgG3, but insignificantly different (P > 0.05) from IgG. Finally, serum specific IgG4 recorded a 100% specificity that was significantly higher than IgG, IgG2, IgG3 (P < 0.01) & IgG1 (P < 0.05). They showed cross reactions with ascariasis, lymphatic filariasis and strongyloidiasis. The results are discussed. PMID- 15880991 TI - Anti-malarial chloroquine stimulate p53-apoptotic pathway in rat hepatocytes. AB - The study investigated rat liver injury produced by anti-malarial dose of chloroquine (CQ) and to elucidate if CQ could induce DNA damage and subsequently p53-dependent apoptosis in rat hepatocytes or not. The effect of anti-malarial CQ on p53-apoptotic pathway of rat liver cells after different time intervals (1, 2 & 3 days) was studied. Apoptosis was assessed in whole liver tissue using ELISA technique and at single cell level using TUNEL technique. The level of expression of p53 was measureed quantitatively by ELISA. Results found that CQ induced DNA damage and rat hepatocytes apoptosis in a time dependent manner. Secondly, p53 expression was associated with CQ administration in time dependent manner. Thus, anti-malarial dose of CQ induces DNA damage and apoptosis of rat liver cells and the expression of p53 could be considered as a normal response of hepatocytes suffering from CQ genotoxic stress. PMID- 15880992 TI - Effects of Solanum nigrum leaves water extract on the penetration and infectivity of Schistosoma mansoni cercariae. AB - A crude water extract of Solanum nigrum leaves was used as a chemical attenuate to Schistosoma mansoni cercariae prior to infection of Swiss female mice. Cercariae were exposed to 2.5, 5, 7.5 and 10 mg/l concentrations of the extract for 30 min. The effect on the ability of cercariae to penetrate mice skin, as well as, effect on schistosome worm burden after 8 weeks of infection were measured. The observed reduction of cercarial penetration was significant at 7.5 and 10 mg/l concentrations (p < 0.001). The mean number of worm burden declined from 28.5 worms/ mouse in untreated group to 4.4 worms/mouse with 7.5 mg/l treatment (p < 0.01). At a concentration of 10 mg/l, mice had no adult worm. The cercarial infectivity, as measured by the proportion of worms recovered in relation to the number of cercariae administrated, decreased with the increase in the extract concentration and was significant at a concentration of 7.5 mg/l (p < 0.01). The number of schistosome eggs in hepatic tissue decreased in treated mice. The reduction in egg count (per gram liver) was significant at 5 mg/l (p < 0.05) and 7.5 mg/I (p < 0.001). The treatment with Solanum water extract had no effect on female fecundity. These data point to Solanum as a promising agent for the control of schistosomiasis. PMID- 15880993 TI - Larvicidal, miracidiacidal and cercaricidal activities of the Egyptian plant Iris pseudacorus. AB - The larvicidal properties of Iris pseudacorus leaves ethanol extract against second-instar larvae of two mosquito species, Culex pipiens and Aedes caspius (Diptera: Culicidae) were studied. It was observed that the larvicidal effect of this extract was dose-dependent. The LC50 values of I. pseudacorus against C. pipiens and A. caspius were 10.36 and 16.43 mg/l within 24 hr, and 7.36 and 10.1 mg/l within 48 hr, respectively. The miracidiacidal and cercaricidal properties of I. pseudacorus extract were directly tested against Schistosoma mansoni miracidia and cercariae and a time-concentration relationship was observed. The concentrations needed to kill all miracidia (LC100) within 5 min., 30 min. and an hr of exposure were 2.7, 1.6 and 0.9 mg/l respectively. The concentrations needed to kill all cercariae (LC100) within 5 min. 30 min. and an hr of exposure were 1.5, 1.0 and 0.6 mg/l respectively. PMID- 15880994 TI - Spot light survey on fresh-water snails of medical importance in Al Fayoum Governorate, Egypt. AB - In a survey carried out during Summer and Autumn of 2004, for snails of medical importance, nine species were recovered. These were Biomphalaria alexandrina, B. glabrata, B. pfeifferi, Bulinus truncatus, B. forskalii, Lymnaea natalensis, Bellamya (=Vivipara) unicolor, Physa acuta and Hydrobia musaensis. Parasitological examination revealed that B. alexandrina, B. glabrata and L. natalensis harboured immature stages of their concerned trematode parasites. Moreover, P. acuta harboured the immature stage of the nematode parasite Parastrongylus cantonensis. PMID- 15880995 TI - Entamoeba histolytica or Entamoeba dispar among children in Gaza, Gaza Strip? AB - Most physicians in Gaza prescribe medicaments for patient's suffering from Entamoeba histolytica/dispar without parasitologic diagnosis. Additionally, stool analysis performed by the routine methods usually reports the species as E. histolytica without con-firmation. In this study, 92 stool specimens were collected and analyzed by wet mount, iron haematoxylin staining, antigen detection of E. histolytica and polymerase chain reaction (PCR). The total number of E. histolytica identified by PCR was 64 (69.6%) that of E. dispar was 21 (22.8%). Mixed infection with both E. histolytica and E. dispar was evident in 7 specimens (7.6%). In the light of these results approximately 30% of suspected clinical amoebiasis cases were negative for E. histolytica. It is recommended to use PCR for diagnosis of stool specimens from patients with E. histolytica/dispar and that treatment should be prescribed for only patients positive for E. PMID- 15880996 TI - Degree of symptoms versus copro-antigen levels in Giardia lamblia infection. AB - A total of 82 out-patients were examined for Giardia copro-antigens and 12 neonate stool samples as control. ELISA had a sensitivity of 100% and specificity of 91.67%. ELISA (O.D.) had neither significant correlation to Giardia cyst count, to stool consistency or presence of blood, mucus or fat in stool, nor to age but positive correlation to the severity of diarrhoea, colic, nausea, anorexia, weight loss, distension and fatigue. Giardia cyst count was higher in cases with loose stool, while ELISA (O.D.) correlated positively with symptoms except constipation and vomiting. The different in clinical outcome of giardiasis can be attributed, partially to strain differences and host resistance. PMID- 15880997 TI - Cholestasis in human fascioliasis in Dakahlia Governorate, Egypt. AB - This study was conducted to assess cholestasis in human fascioliasis. Twenty five patients, passing Fasciola eggs, were compared to twenty age- and sex-matched healthy controls. All were subjected to thorough clinical examination, stool analysis, complete blood picture, liver function tests (AST, ALT, SAP, GGT and total serum bilirubin). Autoimmune and viral hepatitis (HCV & HBV) were excluded. All cases were subjected to radiological examinations (chest x-ray and abdominal ultrasonography). The study revealed significant elevation of liver enzymes in the patients compared to the controls (p < 0.001). Calcular and non-calcular cholecystitis were common findings among patients (32% and 24% respectively). Ascites (4%) and dilated intra-hepatic biliary radicals (32%) were encountered; pleural effusion was detected by chest x-ray in 20% of cases. So, fascioliasis should be considered in the diagnosis of cholestasis in Fasciola endemic areas. PMID- 15880998 TI - Gram-negative bacteria from the camel tick Hyalomma dromedarii (Ixodidae) and the chicken tick Argas persicus (Argasidae) and their antibiotic sensitivities. AB - A total of nine species of gram-negative bacteria were isolated from organs and haemolymph of the hard tick Hyalomma (Hyalomma) dromedarii and the soft tick Argas (Persicargas) persicus. Four species namely Serratia liquefaciens, Stenotrophomonas maltophilia, Klebsiella ornithinolytica and Aeromonas hydrophila were isolated from H. dromedarii and five species namely Rahnella aquatilis, Pseudomonas fluorescens, Enterobacter cloacae, Chryseomonas luteola and Chryseobacterium meningosepticum were isolated from A. persicus. Isolated bacteria were identified using the analytical profile index 20E. Disk diffusion test was carried out on all isolated bacteria to determine antibiotic sensitivity of chloramphenicol, amoxillin/clavulanic acid, neomycin, streptomycin, triplesulphur tetracycline and nitrofurantion. The results were discussed. PMID- 15880999 TI - Immunopathogenic role of IgG antibody and RANTES in house dust mite-induced chronic bronchitis. AB - Based on immunological and clinical examinations, 21 patients were diagnosed as having house dust mite (HDM)-induced chronic bronchitis and classified into three groups according to the clinical presentation of the disease: stable bronchitis, exacerbated bronchitis and asthma on top of bronchitis. Using ELISA, the levels of serum anti-Dermatophagoides farinae and anti-D. pteronyssinus IgG antibodies and plasma RANTES (regulated upon activation, normal T-cell-expressed and secreted; a chemokine with attractive and activator role for eosinophils) were measured in correlation to serum eosinophil cationic protein (ECP, a marker of eosinophil activation and degranulation measured by chemiluminescent immunometric technique). Using immunoblotting, IgG binding components of D. farinae and D. pteronyssinus were determined providing a clue for diagnosis of HDM-induced chronic bronchitis. Significant higher levels of anti-D. farinae and anti-D. pteronyssinus IgG antibodies and RANTES were found in asthmatic group followed by exacerbated chronic bronchitis in comparison to stable bronchitis and control groups. ECP level correlated significantly with IgG and RANTES levels in exacerbated bronchitis and asthmatic groups. The results provided evidence that over expression of IgG and RANTES plays a crucial role, as mediator in immunopathogenesis of HDM-induced chronic bronchitis and as marker of the immunological changes likely responsible for progression of bronchitis to asthma in HDM-sensitive patients yet, RANTES seemed to be an early indicator. Definition of the immunopathogenic role of IgG and RANTES in HDM-induced bronchitis should enable the manipulation of the critical immune response in the hope of establishing new therapies. D. farinae and D. pteronyssinu antigenic bands at > 205 and 205 KDa, respectively, considered together showed 71.4% sensitivity in diagnosis of HDM induced chronic bronchitis and 100% specificity by immuno blotting. PMID- 15881000 TI - Serological diagnosis of Trichinella spiralis in experimentally infected mice. AB - ELISA and IFA tests were used to detect IgG levels in sera of experimentally infected mice with Trichinella spiralis, during a period of 12 weeks post infection. A crude saline extract of muscle larvae was used for application of ELISA technique, while the intact T. spiralis muscle larvae were used as an antigen for IFAT. In both tests, T. spiralis IgG antibodies could be detected one week post infection in sera of all infected mice. ELISA test showed a sensitivity of 100% during the whole period of the experiment, while its specificity was 93.3%. Meanwhile, IFA test revealed a sensitivity of 100% and a specificity of 85%. PMID- 15881001 TI - Biochemical fingerprints of salivary gland for some ticks species in Egypt by SDS polyacrylamide gel electrophresis. AB - SDS-PAGE gel electrophoresis as a sensitive method is used for simultaneously comparing the total protein extracted from salivary glands in some ticks species (Argas percsicus, Hyalomm antolicum excavatum, H. an. antolicum, H. dromedarii, H. impeltatum and Rhipicephalus sanguineus). Results of A. persicus were characterized by 4 bands with molecular weights ranging from 82.76 to 16.076 KD. On the other hand, H. an. excavatum females were characterized by 6 bands with molecular weights ranging from 103.74 to 6.216 KD. H. an. antolicun males were characterized by 5 bands with molecular weights ranging from 79.183 to 2.092 KD. R. sanguineus was characterized by 4 and 8 bands with molecular weights ranging from 88.864 to 6.216 KD and 126.69 to 7.112 KD for male and female respectively. Female H. impeltatum was characterized by 8 bands with a range of molecular weights from 106.96 to 7.829 KD. H. dromedarii was characterized by 4 & 7 bands with ranges of molecular weights from 88.147 to 4.602 KD and 110.20 to 2.630 KD for male and female respectively. PMID- 15881002 TI - Localization of leucine aminopeptidase in the Schistosoma mansoni eggs and in liver tissue from infected mice. AB - Infection with Schistosoma mansoni causes hepatic granuloma formation and fibrosis in response to parasite eggs. The present work localized the leucine aminopeptidase (LAP) in S. mansoni eggs and in liver tissue sections from infected mice. Fresh eggs and livers obtained from infected hamsters were processed and stained with the L-leucine-7-amino-trifluoromethyl-coumarin specific substrate. The L-argnine-7-amino-trifluoro-methylcoumarin and Bestatin (leucine aminopeptidase inhibitor) were used to test the LAP substrate specificity and reactivity. The staining pattern for that enzyme in the egg and liver tissue reflects that the leucine aminopeptidase is a major egg constituent distributed in nearly all the egg except the spine. The control substrates confirmed the substrate broad specificity of LAP. In conclusion, the LAP enzyme is a major egg antigen and the target antigen for the antipathology vaccine development studies. PMID- 15881003 TI - Hydatidosis among man and animals in Assiut and Aswan Governorates. AB - A survey for hydatidosis among the slaughtered animals at Assiut and Aswan abattoirs, over one year showed that hydatid cyst camels was 107 (7.67%) out of 1395, but no infection in cattle and buffaloes. The lung was more involved than liver in camels. The fertile hydatid cysts in camels were 60.41% and 54.23% in Assiut and Aswan Governorates respectively. The hydatid cysts recorded in Assiut and Aswan Gs during Summer (15.78%, 6.34%) or Autumn (12.0%, 7.83%) was higher than during Winter (10.58%, 3.03%) or Spring (10.52%, 5.18%). Fresh fertile hydatid cysts protoscolices were recovered from lungs of infected camels, slaughtered at Bani-Adi (Assiut) abattoir were orally given to experimental dogs. All dogs developed Echinococcus worms mainly in the small intestine proximal third. The levels of echinococcosis by IHAT were estimated in 100 patients with acute & chronic hepatic diseases in Assiut and in Aswan Gs, showed 5 (10.0%) positive reactions. PMID- 15881004 TI - Early post-treatment immunoglobulin profile in human schistosomiasis. AB - In a trial at determining the most relevant immunoglobulin isotype that could reflect success of praziquantel treatment, an ELISA using soluble egg antigen (SEA) was applied on sera of Egyptian patients suffering from active intestinal schistosemiasis without hepatic complications, determining the levels of IgE, IgA, IgM, IgG1, IgG2, IgG3 and IgG4 raised against the SEA, both, pre- and early post-treatment. The positive results obtained to all anti-SEA immunoglobulin isotypes before treatment support the usefulness of this technique in the diagnosis of schistosomiasis. Except for IgG3 subclass, a statistically significant correlation was found between egg output-reflecting intensity of infection- and the different immunoglobulin levels, especially anti-SEA IgG4. When repeating the assay 5-6 months after treatment, the immunoglobulin levels showed either a rise (in case of IgE) or a drop (in case of IgA, IgM & IgG1-4), all of statistical significance, yet, IgG1-4 were still positive. So, ELISA could not give a definite indication of cure after anti-bilharzial treatment. IgE, IgG2 and IgG4 were revealed to be the most significant immunoglobulin isotypes at the post-treatment level, both statistically and due to their implications on resistance/ susceptibility to re-infection and also due to the correlation of IgG4 with the tendency to develop periportal fibrosis. Conclusively, although not having defined a particular Ig isotype as marker for cure, yet it exposed the urge for early post-treatment determination of IgE and IgG4 isotypes, which could serve as markers for picking up high risk patients susceptible to reinfection or liable to develop bilharzial periportal fibrosis, and who might benefit from a second course of specific treatment. PMID- 15881005 TI - Human fascioliasis in some areas in Cairo and Giza Governorates, Egypt. AB - This study comprised 462 clinically suspected cases of fascioliasis. Stool examination revealed positive 11 cases for Fasciola eggs (2.38%). Serum samples were obtained for serodiagnosis from 133 suspected cases from Cairo governorate (G) and 149 from Giza G. Using IHAT, positive results were reported in 12.03% of suspected cases in Cairo G and 11.41% in Giza G. For ELISA 15.03% and 18.12% respectively were positive. Out of 462 clinically suspected cases, 58 patients were liable to be infected with Fasciola depending on the presence of Fasciola eggs in stool and/or a positive serological result with either IHAT (56.9%) or ELISA (81.03%) or both (48.28%). Serological examination of sera of cases shedding eggs revealed that one case was positive for ELISA, 4 cases were positive for IHAT and ELISA, whereas 5 cases were serologically negative. Serum from one case with Fasciola eggs in stool could not be obtained for serological diagnosis. Clinical picture of infected cases indicated that 82.76% were suffering from abdominal pain, 43.1% palor and digestive disturbances, and 25.86% fever. On physical examination, 46.55% revealed hepatomegaly. Eosino-philia was observed in 70.69% of the patients. Statistically, no significant difference (P. > 0.05) was observed among patients in both governorates regarding age, sex, clinical manifestations, detection of eggs in stool, hepatomegaly, and serological tests. However, eosinophilia was significantly higher (P < 0.05) in cases from Giza G. PMID- 15881006 TI - Cerebrospinal echinococcosis: serodiagnosis using different hydatid cyst fluid antigens. AB - Cerebrospinal hydatid disease diagnosis may impose some problems, as ultrasonography is not applicable and serology may not detect the low antibody titre often associated with intracranial or orbital cystic echinococcosis. Serological tests were performed on 14 cases with intracranial or spinal cystic lesions, out of which 9 were cases of hydatidosis, in addition to 26 cases with other parasitic diseases as detected by stool examination. The sensitivity of all tests and antigens used did not exceed 6 out of the 9 positive cases, and achieved by the Eg1 and Eg2 when applied in the ELISA. The specificity of this technique using the two antigens was 92.3%& 76.9%, respectively. Semi-purified Em1 and purified 44 KDa used in the ELISA, crude Eg1 used for the CIEP and the crude commercial antigen of the IHAT, all gave lower sensitivities than the former two antigens, yet their specificities amounted to 100%. In conclusion, for diagnosis of cerebrospinal hydatidosis the home-prepared Eg1 antigen is recommended in the ELISA system, as it is relatively easily prepared from available resources to be supplemented with radio-imaging techniques especially Magnetic Resonance Imaging and/or MR spectroscopy, the latter being very helpful in clearly differentiating various types of intracranial cysts. PMID- 15881007 TI - The nematode Goezia sp. (Anisakidae) from Bagrus bayad (Osteichthyes) from Egypt. AB - The present study describes nematode infection in the freshwater fish Bagrus bayad (Osteichthyes) collected from the Red Sea at Suez Canal Zone. Of 32 examined fish, 21 (65%) were infected with Goezia sp. (Nematoda: Family: Anisakidae) with mean ntensity of 4.1 parasites. The nematode present here differs from Goezia braziliensis, G. breviaeca and G. intermedia, as well as from other species described. The main difference was in the greater number of pre anal papillae in males compared to G. braziliensis. The present parasite also differs from G. braziliensis in spicule length and distance of vulva from the anterior extremity. PMID- 15881008 TI - Ornithonyssus (Acari: Macronyssidae) mite dermatitis in poultry field-workers in Almarg, Qalyobiya governorate. AB - Cutaneous manifestations of bird and rat mite infestation in man are not easily recognized by physicians or patients. Clinical signs and symptoms are developed secondary to bites of mites that have infested rats, domestic poultry or birds nesting in or near human habitation and comes into contact with man. This study details 4 cases of pruritic dermatitis developed in four field workers in poultry farms in Al-Marg district, Qalyobia governorate, Egypt. The zoonotic species of Ornithoyssus sp., (Family Macronyssidae) was isolated from all samples collected from patients' habitat and the role played by Ornithonyssus mites in causing dermatitis in man was discussed. PMID- 15881009 TI - Biochemical modifications induced in rabbits by Schistosoma mansoni antigens and the beneficial effect of carnosine treatment. AB - This immunological study involved individual injection of the three Schistosoma mansoni antigens (Ags). soluble egg antigen (SEA), cercarial antigen preparation (CAP) or soluble worm antigen preparation (SWAP) in three rabbits groups (Ag). respectively. Three other groups each received the same specific antigen conjoined with administration of L-carnosine (Ag-C). Determination of three hepatic parameters and ten serum proteins was done. These were total protein, glycogen content and glycogen phosphorylase b activity of liver as well as serum total protein and nine protein fractions [alpha2-macrglobulin; beta galactosidase; phosphorylase b; serum albumin; fumarase; carbonic anhydrase; beta lactoglobulin; alpha-lactalbumin and aprotinin]. Conjoined carnosine treatment produced numerous variations. SEA-I-C group presented sex decreased parameters. In CAP-I-C animals hepatic glycogen content was increased while phosphorrylase b activity was decreased as well as seven the concentration of serum parameters; total serum protein, alpha2-macroglobulin, phosphorylase b, albumin, fumarase, carbonic anhydrase, alpha-lactalbumin and aprotinin. In SWAP-I-C group the concentration of only one fraction was decreased; carbonic anhydrase. In batch A both the Ags. of the egg and cercaria, developmental stages having transient residence in the animal host, showed more affection by the specific Ag. Although, carnosine modified the results of all the three groups in batch B yet, its effect on both the egg and cercaria Ags. was still more than that of worm. PMID- 15881010 TI - On the main reservoir host of Fasciola in Dakahlia Governorate, Egypt. AB - In the present work, a total of 4180 sheep were obtained from different localities at Dakahlia Governorate. One hundred and eighty male sheep were investigated for Fasciola worms (the adult and juvenile forms) at Mansoura governmental abattoir. Four hundred grazing sheep were screened by the coprological examination from ten centers. The mean number of eggs per gm in a group of ten sheep ranged from 2.9-10.8 egg/gm. The number of worms ranged from 5 84. In the Mansoura abattoir, it was found that 37 sheep were infected (20.56%). Fasciola worms were recovered from the livers in 100%, common bile ducts in 60%, gall bladder in 80%, two sites in 50% and three sites in 40% of infected biliary tracts. According to the coprological examination, the infection rate was the highest in El-Manzalla Center (15.75) and the lowest was in Mansoura Center (3.25%). According to the prevalence and egg output results, it is concluded that sheep are the main reservoir host for fascioliasis among the edible animals in Dakahlia Governorate. However, examination of the non-edible animals and the domestic and wild rodents are ongoing and will be published soon elsewhere. PMID- 15881011 TI - Effect of endoscopic injection sclerotherapy and variceal ligation on development of portal hypertensive gastropathy and duodenopathy. AB - The portal hypertensive gastropathy (PHG) and duodenopatby (PHD) define a wide spectrum of diffuse macroscopic lesions that appear in the gastric mucosa of patients with portal hypertension. Histologically, the lesions corresponded to dilated vessels in the mucosa and submucosa in the absence of erosions or inflammation. This work studied the effect oesophageal varices (OV) eradication either by endoscopic sclerotherapy and/or band ligation on PHG and PHD. Sixty portal hypertensive patients were classified into three groups: G.I- 20 patients with no history of bleeding or endo-therapy as control. G.Il- 20 patients with complete oesophageal eradication of OV by endoscopic sclerotherapy (EVL). G.III- 20 patients with complete oesophageal eradication by endoscopic sclerotherapy (ES). PHG was 55% in controls, 70% in EVL and 65% in EVL. PHG was 20% in controls, 35% in EVL and 35% in ES. PHG and PHD increased after complete eradication of the varices either by ES or EVL, but without statistical significant differences. PMID- 15881012 TI - Toxicological deterioration of some plant extracts on Fannia canicularis (Diptera: Muscidae). AB - The early third instar larvae of Fannia canicularis were treated with various concentrations of Rumex dentatus, Protulaca deracea and Piper cubebae extracts. The Rumex dentatus was more toxic than Protulaca deracea and Piper cubebae. The extract mixture of Rumex dentatus and Piper cubebae induced a high synergistic action against the treated larvae, but the mixture of Protulaca deracea and Piper cubebae indicated an antagonistic effect. The reproductive potential of treated insects together with the biochemistry of their ovaries and testes were determined. The results indicated that the tested mixtures had delayed effects represented by the decreased the ovarian and testicular protein and the disturbance of their amino acid contents. This might be due in part to the interference of these mixtures with gonadal development, which affected negatively the fecundity and fertility of adults. PMID- 15881013 TI - Cross-protection induced by cross-reactive antigen against Fasciola gigantica and Trichinella spiralis infections. AB - Trichinella spiralis /Fasciola gigantica cross-reactive fraction was purified from T. spiralis larval extract by affinity column chromatography in which CNBr Sepharose 4B was coupled with F. gigantica antibodies. The fraction consisted of six polypeptides of 191KDa, 178KDa, 149KDa, 106KDa, 101KDa and 32 KDa as revealed by SDS-PAGE. Analysis of the free amino acids of the fraction revealed 17 amino acids with high proportions of tyrosine and glutamic. Immunization of rabbits subcutaneously with the cross-reactive fraction in Freund's adjuvant followed by challenge with F. gigantica metacercariae resulted in reduction in worm burdens reached to 47.8%. While immunization of rats with the same fraction in Freund's adjuvant followed by infection with T. spiralis larvae resulted in reduction in worm count reached to 74%. IgG antibody response in rabbits increased due to immunization to reach its maximum value at the time of infection and then decreased gradually up to the end of the experiments. But. remained higher than the level in non vaccinated control animals. In rat sera, IgG level increased due to vaccination but the level recorded its optimum value one week post infection and then decreased. Thus the cross-reactive antigen proved cross-protection with the protection inducing capability against both diseases. PMID- 15881014 TI - Microscopic study on Sarcocystis moulei from sheep and goats in Saudi Arabia. AB - The present investigation revealed that the role of natural infection with Sarcocystis in sheep was 84% in Niemy, 76% in Najdy, 89% in Sawakny sheep and 77% in goats slaughtered in Riyadh, Saudi Arabia. The distribution of infection among the examined organs was varied from the sheep strain to goats. Recorded muscle cysts were macroscopic 8-18% (mean 13.5%) and microscopic 73-85% (mean 79%). Macroscopic cysts were usually detected on the oesophagus. EM showed an ultrastructural similarities in the cyst walls and their parasite contents (metrocytes and merozoites) for all macroscopic cysts examined from sheep and goats. These were also enclosed by a secondary cyst wall. The primary cyst wall had many cauliflower-like protrusions supported by many fibellar structures, while the ground substance underneath them contained many dark, dense granules and usually extended into the cyst cavity dividing it into many compartments containing the parasites. The cyst parasites were the peripheral globular to spherical metrocytes, which were divided by endodyogeny and the central banana shaped merozoites. Both metrocytes and merozoites had all characteristics of the Apicomplexa. Experimental transmission of Sarcocystis to the definitive host, revealed that kittens were the only suitable final host. Thus, gamogony and zygote formation (fertilization) occurred 2-8 days post infection (p.i.) while sporulation took place within this final host 13-15 days p.i., where sporocysts were passed within faeces of infected kittens at that time. Also, the study indicated that the prepatent period was 13-15 days, while the patent period was 37-42 days. PMID- 15881015 TI - Comparative study of the hepatotoxic, genotoxic and carcinogenic effects of praziquantel distocide & the natural myrrh extract Mirazid on adult male albino rats. AB - Praziquantel (PZQ) is widely and effectively used in the control of bilharziasis which constitutes a major endemic health problem in Egypt. However, recent studies recommended that the drug must be re-evaluated because of its potential carcinogenicity and genotoxicity. Mirazid is a new natural anti-schistosomal drug formed of myrrh extract and considered to be a safe drug. This work was conducted to evaluate and compare hepatotoxic, genotoxic and carcinogenic effects of PZQ and Mirazid on adult male albino rats by assessment of serum levels of ALT, AST and bilirubin, histopathological study of the liver and cytogenetic study of bone marrow cells. 100 adult male albino rats were equally divided into 4 groups: (I): negative control, (II): control rats received distilled water, (III): received weekly single oral dose of PZQ (1500 mg/kg) for 6 weeks, (IV): received daily oral dose of Mirazid (500 mg/kg) for 6 weeks. At the end of the study 10 rats of each group were investigated by assessment of the levels of AST, ALT, & Bilirubin. After scarification, liver sections were examined by light microscopy. Another 10 rats of each group were submitted to cytogenetic examination. It was found that praziquantel induced a significant increase in the mean values of AST, ALT and bilirubin with areas of hyaline degeneration, fatty changes, dysplasia and necrosis in the liver sections. It also induced a significant increase in the incidence of chromosomal aberrations as polyploidy, fragment, deletion and ring chromosome as compared with control group. Mirazid induced a non significant increase in the mean values of AST, ALT and bilirubin, with a normal hepatic tissue, and a non significant increase in the incidence of chromosomal aberrations, as compared with the control group. On comparing both drugs, praziquantel induced a significant hepatotoxic, genotoxic and carcinogenic effects. It was concluded that, Praziquantel is considered to be a hepatotoxic, genotoxic and carcinogenic drug. On the other hand, Mirazid seemed to be a safe and promising antiparasitic drug, free from hepatotoxic, genotoxic and carcinogenic effects. PMID- 15881016 TI - Megaselia scalaris causing human intestinal myiasis in Egypt. AB - Megaselia scalaris is a worldwide distributed insect of medical importance. In a laboratory-based study, stool samples with undefined maggot infestation were examined and the presence of M. scalaris maggots was confirmed. Binocular stereo microscopy was used for identification of the maggots. Larvae were allowed to develop into adults onto a human stool culture. The larvae and the emerged flies were identified using standard keys. This may be the first report of M. scalaris as a causative agent of human myiasis in Egypt. Details of the third instar larva, pupa and adults were given. PMID- 15881017 TI - Zoonotic toxoplasmosis in chicken. AB - Modified agglutination test (MAT) was applied to 150 chicken sera (90 were farm bred and 60 house-bred chickens). The prevalence of Toxoplasma antibodies was 18.7%. In house-bred chickens positive cases were 18 out of 60 (30.0%), while in farm-bred chickens positive cases were 10 out of 90 (11.1%). Histopathological sections of the seropositive chickens showed tissue lesions which were most likely to be induced by T. gondii. Tissue cysts of T. gondii were demonstrated in the liver, brain, heart and skeletal muscles of 22 (78.6%) out of 28 positive chickens. These tissue cysts were found mostly in the brain of seropositive chickens. Epidemiological implications of these data for public health significance were discussed. PMID- 15881018 TI - Dipterous flies species and their densities in fourteen Egyptian Governorates. AB - The present study focused on dipterous flies, distribution and densities in 14 Egyptian Governorates. The abundance and distribution of flies was carried out in three phases. The first was in 1999 in 5 governorates, Qalyobia, Menoufia, Fayium, Behiera & Assuit included species of the families: Calliphoridae, Chloropidae, Drosophilidae, Milichiidae, Muscidae, Otitidae, Piophilidae, Sarcophagidae, Sepsidae, Sphaeroceridae and Syrphidae. The second was in 2000 in another five Governorates, Kafr-ElSheikh, Giza, Menia, Aswan and Sharkia included sp. of the families: Calliphoridae, Chloropidae, Drosophilidae, Milichiidae, Muscidae, Otitidae, Piophilidae, Sarcophagidae, Sepsidae, Sphaeroceridae and Syrphidae. The third phase was in 2001 in 4 Governorates: Kena, El-Wady El Gedied, Da-kahlia and South Sinai included species of the families: Calliphoridae, Chloropidae, Drosophilidae, Milichiidae, Muscidae, Otitidae, Phoridae, Piophilidae, Sarcophagidae, Sepsidae, Sphaeroceridae and Syrphidae. Musca domestica was the most abundant sp. found throughout the work. The results are a step in drawing an update map for insect-vectors to help in the national control programs. PMID- 15881019 TI - Treatment of sheep fascioliasis with Macca Mur. PMID- 15881020 TI - Evaluation of environmental burdens associated with sewage treatment processes using life cycle assessment techniques. AB - Life cycle assessments of a range of wastewater treatment processes have been undertaken. At lower consent standards rotating biological contactors and biological filters exhibit lower environmental burdens for the required functionality than other process options. However, at tighter consents this distinction becomes less clear. Attempts to reduce environmental burdens should focus on operational energy requirements (which dominate burden profiles and, where appropriate, on synthetic materials used for bio-mass growth). However, differences between dry and operational weight, and the consequent size of necessary foundation structures are likely to limit opportunities for reducing overall burden profiles through material substitution. Comparison of processes across population equivalents suggests economies of scale may favour adopting larger scale plant, although there are additional factors yet to be included in this analysis. Furthermore, adopting different waste disposal routes during the demolition phase is unlikely to have a distinct impact on burden profiles. PMID- 15881021 TI - Application of urea based SNCR system in the combustion effluent containing low level of baseline nitric oxide. AB - Selective Non-Catalytic Reduction (SNCR) of nitric oxide has been studied experimentally by injecting aqueous urea solution with and without additive in a pilot-scale diesel fired tunnel furnace at 3.4% excess oxygen level and with low ppm of baseline NO(x) ranging from 65 to 75 ppm within the investigated temperature range. The tests have been carried out using commercial grade urea as NO(x) reducing agent and commercial grade sodium carbonate as additive. The furnace simulated the small-scale combustion systems, where the operating temperatures are usually in the range of about 973 to 1323 K and NO(x) emission level remains below 100 ppm. With 5% plain urea solution, at Normalized Stoichiometric Ratio (NSR) of 4 as much as 54% reduction was achieved at 1128 K, whilst in the additive case the NO(x) reduction was improved to as much as 69% at 1093 K. Apart from this improvement, in the additive case, the effective temperature window as well as peak temperature of NO(x) reduction shifted towards lower temperatures. The result is quite significant, especially for this investigated level of baseline NO(x). The ammonia slip measurements showed that in both cases the slip was below 16 ppm at NSR of 4 and optimum temperature of NO(x) reduction. Finally, the investigations demonstrated that urea based SNCR is quite applicable to small-scale combustion applications and commercial grade sodium carbonate is a potential additive. PMID- 15881022 TI - Enhanced coagulation of disinfection by-products precursors in Istanbul water supply. AB - During the chlorination of drinking water, chlorine reacts with natural organic matter (NOM) to produce disinfection by-products (DBPs), such as trihalomethanes (THMs) and haloacetic acids (HAAs), which are believed to be harmful to human health. Enhanced coagulation is a DBPs precursor treatment technique with the objective of removing total organic carbon (TOC) to control DBPs formation in water. Scientifically, the removal of organic matter by coagulation depends on the TOC concentration, the chemical nature of the NOM, coagulant type, coagulant dosage, and coagulation pH. Currently, water treatment plants are practicing enhanced coagulation to further increase DBPs precursor removal. The focus of this study was to investigate enhanced coagulation of Terkos Lake Water (TLW) of Istanbul City. In this study, jar test experiments were conducted on TLW source to determine the effectiveness of enhanced coagulation for removal of DBPs surrogate parameters of total organic carbon (TOC), ultraviolet absorbance (UV(254)), and THM formation potential (THMFP). Jar tests results indicated that enhanced coagulation can increase the removal of DBPs precursors. To evaluate the coagulation performances, two different coagulants, aluminum and iron salts were used at different pH values to determine optimal coagulation conditions for surrogate parameters removal quantity. PMID- 15881023 TI - Continuous sorption of copper and cobalt by crab shell particles in a packed column. AB - The ability of crab shell to remove copper and cobalt from aqueous solutions was examined in an up-flow packed column. The experiments were conducted to study the effect of important design parameters such as bed height and flow rate. At a bed height of 25 cm, metal uptake capacity of crab shell for copper and cobalt was 52.07 and 20.47 mg g(-1) respectively. In addition, the results obtained at different flow rates indicated that an increase in flow rate decreased the sorption performance of the crab shell. The Bed Depth Service Time model and the Thomas model were used to analyze the experimental data and the model parameters were evaluated for copper and cobalt sorption. The column regeneration studies were carried out for five sorption-desorption cycles. Loss of sorption performance was observed as the cycles progressed, indicated by a shortened breakthrough time and a broadened mass transfer zone. However, crab shell maintained a good metal sorption capacity for all the five cycles. The elutant used for the regeneration of the crab shell, 0.01 M EDTA at pH 3.5 adjusted using HCI, exhibited elution efficiencies greater than 98%. PMID- 15881024 TI - Bioleaching of Cr from tannery sludge: the effects of initial acid addition and recycling of acidified bioleached sludge. AB - A technologically feasible process called bioleaching was used to remove Cr from tannery sludge with Acidithiobacilli TS6 bacteria in this study. The buffering capacity of tannery sludge and municipal sewage sludge were compared. The effects of initial sulfuric acid addition on bioleaching process and influences of recycle rate of acidified bioleached sludge on subsequent bioleaching reaction were examined. The quantities of initial sulfuric acid addition ranged from 0 to 0.185 ml g(-1) dry sludge. Recycle rate (V(beleached/V(total)) was controlled at 1/3, 1/2, 3/5 and 2/3. We found that tannery sludge had a higher buffering capacity than sewage sludge. The results showed that there was an increase in the rates of pH reduction and Cr solubilization with the increase of initial sulfuric acid addition. The suitable quantity of sulfuric acid addition was 0.185 ml g(-1) dry sludge at the beginning of the first bioleaching process. After 10 days of bioleaching, pH reached 1.5 and almost 100% of Cr could be leached in this case. Besides, an evidently dividing point pH 2.0 was found to be the threshold value for considerable Cr solubilization during the leaching reaction. Recycle experiments confirmed that the application of the method by recycling acidified sludge to launch subsequent bioleaching is feasible. The optimal recycle rate of acidified sludge for consecutive leaching of Cr from tannery sludge is 2/3. PMID- 15881025 TI - The formation, role and removal of NO3-N during corona discharge in air for phenol removal. AB - Pulsed corona discharge carried out in the gas phase has been shown to be a low cost solution for wastewater treatment. In this study, the results show that the nitrogen dissociated during corona discharge in the gas phase formed nitric acid in solution, which resulted in a drop in solution pH. The phenol removal could be described in accordance with a first-order reaction in a basic buffer solution, but not in initial neutral and basic non-buffered solutions. This indicates that basic conditions facilitate phenol removal, whilst the formation of nitric acid in solution resulted in decrease in the phenol removal rate. This study also shows that most of the oxidants used for phenol removal came mainly from the dissociation of water molecules at the surface of the liquid phase and/or the gas phase very close to the liquid surface in discharging conditions, not from the gas dissociation in the bulk gas phase, and therefore, it is more effective to keep the solution basic than to change the component of gas in the reactor when removing phenol from water using corona discharge. PMID- 15881026 TI - Effect of moisture content on fed batch composting reactor of vegetable and fruit wastes. AB - Vegetable and fruit wastes mixed with sawdust were composted in a laboratory scale reactor by controlling the waste feeding rate at 21 kg m(-3) day(-1) and aeration rate at 10.6 l m(-3) min(-1). The effects of initial moisture content on organic matter degradation and process performance of fed batch composting were investigated. The absolute amount of removal, removal percentage, and removal rate of dry mass obtained were substantially different among the initial moisture contents. The rapid rise of moisture content and the lowest absolute amount of removal observed were achieved in the 50% condition. The initial moisture content yielding the largest absolute amount of removal in both feeding and curing stage was 30% whereas the removal percentage and rate constant of waste decomposition were highest in the 50% condition. Examined by traditional soil physics method, the moisture content at 50-55% was suitable for satisfying the degree of free air space (65-70%) of compost during the fed batch composting. Most degradable organic matter was mainly consumed in the feeding stage as indicated by a higher removal rate of dry mass in all cases. It is recommended that the initial moisture content of 30% and mode of aeration and agitation should be adopted for achieving practical fed batch composting of vegetable and fruit wastes. The study also demonstrated that the composting kinetics of vegetable and fruit wastes mixed with sawdust can be described by a first order model. PMID- 15881027 TI - A simple biofilter for treatment of pig slurry in Malaysia. AB - On commercial pig production farms in South East (SE) Asia, the liquid effluent is often discharged into rivers. The discharge is a hazard to the environment and to the health of people using water from the river either for consumption or for irrigation. Therefore, a simple percolation biofilter for treatment of the liquid effluent was developed. Pig slurry was treated in test-biofilters packed with different biomass for the purpose of selecting the most efficient material, thereafter the efficiency of the biofilter was examined at farm scale with demo biofilters using the most efficient material. The effect of using "Effective Microorganisms" (EM) added to slurry that was treated with biofilter material mixed with Glenor KR+ was examined. Slurry treatment in the test-biofilters indicated that rice straw was better than coconut husks, wood shavings, rattan strips and oil palm fronds in reducing BOD. Addition of EM and Glenor KR+ to slurry and biofilter material, respectively, had no effect on the temperature of the biofilter material or on the concentrations of organic and inorganic components of the treated slurry. The BOD of slurry treated in test biofilters is reduced to between 80 and 637 mg O2 I(-1) and in the demo biofilter to between 3094 and 3376 mg O2 l(-1). The concentration of BOD in the effluent is related to the BOD in the slurry being treated and the BOD concentration in slurry treated in test biofilters was lower than BOD of slurry treated in demo biofilters. The demo biofilter can reduce BOD to between 52 an 56% of the original value, and TSS, COD (chemical oxygen demand) and ammonium (NH4+) to 41-55% of the original slurry. The treated effluent could not meet the standards for discharge to rivers. The composted biofilter material has a high content of nitrogen and phosphorus; consequently, the fertilizer value of the compost is high. The investments costs were 123 US dollar per SPP which has to be reduced if this method should be a treatment option in practise. PMID- 15881028 TI - Physico-chemical treatment of wastewater from clusters of small scale cotton textile units. AB - Small scale industries can not own individual wastewater treatment facility due to non-availability of land and skilled manpower for operation and maintenance of wastewater treatment plants. A centralized wastewater treatment facility for clusters of small scale industries is appropriate. This concept is gaining popularity in recent years. In India, various textile process operations are undertaken by individual small scale units. The wastewater generated at these units is conveyed to a common effluent treatment facility comprising of equalization, flocculation-clarification, activated sludge process, secondary clarification and finally discharge into inland surface water bodies. The wastewater from small scale cotton textile processing units was highly coloured and alkaline with average BOD and COD concentration of 205 and 790 mg l(-1), respectively. Due to the presence of several dyes, particularly reactive dyes, the biological treatment is often found less effective. Therefore, applicability of various physico-chemical treatment methods needs to be investigated in pursuit of an alternative to biological treatment of textile wastewater. A physico chemical treatment scheme, involving chemical coagulation-sedimentation, dual media filtration, activated carbon adsorption followed by chemical oxidation was investigated in this paper. The quality of final treated wastewater in terms of BOD and COD was 18-24 and 230-240 mg l(-1), respectively through this scheme. A scheme of treatment comprising coagulation-sedimentation, dual media filtration, activated carbon, chemical oxidation may be considered as an alternative to biological treatment of textile wastewater. PMID- 15881029 TI - The influence of pH on hydrolysis and acidogenesis of kitchen wastes in two-phase anaerobic digestion. AB - Batch and semi-continuous experiments were carried out to determine the effect of pH on hydrolysis and acidogenesis of kitchen wastes in the two-phase anaerobic digestion. The results of the batch experiment showed pH adjustment could improve both hydrolysis and acidogenesis rates of kitchen wastes. Compared with pH at 5, 9 and 11, pH 7 provided an optimum working condition for anaerobic digestion of kitchen wastes. At pH 7, about 86% of the total organic carbon (TOC) and 82% of chemical oxygen demand (COD) were solubilized and the maximum volatile fatty acid (VFA) concentration of 36 g l(-1) was achieved on the fourth day. The total VFA yield was 0.27 g (g total solid (TS))(-1), which almost doubled the yield without pH adjustment (0.15 g (g TS)(-1). The acidified products with lower concentration of lactic acid were more favorable to the subsequent methanogenesis. At pH 7, most of the protein was degraded into ammonia nitrogen (NH4(+)-N), resulting in additional buffer of acidified liquid. According to this, a novel method of pH adjustment applying NH4(+)-N buffer was established. The semi-continuous experiment indicated that adjusting pH to 7 in the acidogenic stage in the two phase anaerobic digestion process would increase both TS loading rate (16g l(-1) d(-1)) and recovery of biological energy (520 ml methane (CH4) (g TS)(-1)). PMID- 15881030 TI - A Fenton-like oxidation process using corrosion of iron metal sheet surfaces in the presence of hydrogen peroxide: a batch process study using model pollutants. AB - This study evaluates a new method for chemically destroying organic pollutants in wastewater using spontaneous corrosion of iron metal sheet surfaces in the presence of hydrogen peroxide. Model pollutants (phenol and benzoic acid) were degraded in batch experiments to investigate which parameters affected the process performance. Iron metal sheet surfaces spontaneously corrode under acidic conditions producing iron species (mainly ferrous ions) dissolved in aqueous solution, which react with hydrogen peroxide via the Fenton reaction. In order to optimise the oxidation system, several factors (pH, H2O2 dosage, initial concentration of organic substances) affecting corrosion of the iron metal sheet surface were investigated. Total iron concentration in solution was investigated with different dosages of H2O2 (100 mg l(-1), 1000 mg l(-1) and 1900 mg l(-1)) at different pH values (1.5, 2.5 and 3.0). Iron corrosion increased with the decrease of pH. The addition of H2O2 resulted in a significant increase of iron corrosion. Organic substances also had a marked effect with, for instance, the presence of phenol or benzoic acid resulting in a considerable increase of iron corrosion. In contrast, the absence of either hydrogen peroxide or iron metal brought no change in total organic carbon (TOC). In order to obtain the most effective combination of parameters for TOC removal of phenol solution, experiments were conducted under varied conditions. The experimental results showed that there is an optimum pH requirement (in this work, 2.5). The factors affecting the TOC removal are discussed and the oxidation mechanisms leading to mineralization of organic substances are proposed. PMID- 15881031 TI - Learning to manage leprosy after 2005: preserving critical knowledge and exploiting new technology. PMID- 15881032 TI - Challenges presented by nerve damage in leprosy. AB - The basis of nerve damage in leprosy is the unique tendency of Mycobacterium leprae to invade Schwann cells. alphaBeta-Dystroglycan on the basement membrane of Schwann cells binds to laminin alpha2, in turn binding to receptors on the M. leprae surface, comprising a histone-like protein and phenoglycolipid-1. When nerve damage during reversal reactions was found to be associated with an abrupt increase in delayed type hypersensitivity against M. leprae antigenic determinants released from Schwann cells, it suggested that the nerve is damaged as an innocent bystander during the immune response. This strongly influenced the introduction of therapy based on immunosuppression combined with continued anti mycobacterial medication. Lysis of Schwann cells presenting M. leprae antigenic determinants by activated CD4+ T cells and interaction of M. leprae with Toll like receptors on Schwann cells are additional mechanisms implicated in nerve damage. Persistence of M. leprae antigen in local lesions after regular multiple drug therapy (MDT) is an important risk factor for late reactions. In spite of significant advances in the provision of MDT globally, early diagnosis, together with effective treatment of the disease and associated nerve damage at initial presentation remains a major challenge for the health services. Reduced prevalence as a result of MDT should not be taken to indicate that the challenges of leprosy control are diminished as long as nerve damage is not controlled and new case detection rates are not declining. PMID- 15881033 TI - The INFIR Cohort Study: investigating prediction, detection and pathogenesis of neuropathy and reactions in leprosy. Methods and baseline results of a cohort of multibacillary leprosy patients in north India. AB - The aim of this study was to find predictors of neuropathy and reactions, determine the most sensitive methods for detecting peripheral neuropathy, study the pathogenesis of neuropathy and reactions and create a bank of specimen, backed up by detailed clinical documentation. A multi-centre cohort study of 303 multibacillary leprosy patients in Northern India was followed for 2 years. All newly registered MB patients requiring a full course of MDT, who were smear positive and/or had six or more skin lesions and/or had two or more nerve trunks involved, were eligible. A detailed history was taken and physical and neurological examinations were performed. Nerve function was assessed at each visit with nerve conduction testing, warm and cold detection thresholds, vibrometry, dynamometry, monofilaments and voluntary muscle testing. Because the latter two are widely used in leprosy clinics, they were used as 'gold standard' for sensory and motor impairment. Other outcome events were type 1 and 2 reactions and neuritis. All subjects had a skin biopsy at registration, repeated at the time of an outcome event, along with a nerve biopsy. These were examined using a variety of immunohistological techniques. Blood sampling for serological testing was done at every 4-weekly clinic visit. At diagnosis, 115 patients had an outcome event of recent onset. Many people had skin lesions overlying a major nerve trunk, which were shown to be significantly associated with an increased of sensory or motor impairment. The most important adjusted odds ratios for motor impairment were, facial 4.5 (1.3-16) and ulnar 3.5 (1.0-8.5); for sensory impairment they were, ulnar 2.9 (1.3-6.5), median 3.6 (1.1-12) and posterior tibial 4.0 (1.8-8.7). Nerve enlargement was found in 94% of patients, while only 24% and 3% had paraesthesia and nerve tenderness on palpation, respectively. These increased the risk of reactions only marginally. Seven subjects had abnormal tendon reflexes and seven abnormal joint position sense. In all but one case, these impairments were accompanied by abnormalities in two or more other nerve function tests and thus seemed to indicate more severe neuropathy. At diagnosis, 38% of a cohort of newly diagnosed MB leprosy patients had recent or new reactions or nerve damage at the time of intake into the study. The main risk factor for neuropathy found in this baseline analysis was the presence of skin lesions overlying nerve trunks. They increased the risk of sensory or motor impairment in the concerned nerve by 3-4 times. For some nerves, reactional signs in the lesions further increased this risk to 6-8 times the risk of those without such lesions. Patients with skin lesions overlying peripheral nerve trunks should be carefully monitored for development of sensory or motor impairment. PMID- 15881034 TI - Factors contributing to delay in diagnosis and start of treatment of leprosy: analysis of help-seeking narratives in northern Bangladesh and in West Bengal, India. AB - The objective of our research was to identify factors contributing to delay in diagnosis and start of treatment in leprosy, focussing on patients' narratives of help-seeking behaviour. Our research took place in Purulia, West Bengal, India and in Nilphamari, northern Bangladesh. Between January and August 2000, we conducted semi-structured interviews with 104 patients that explored each individual's narrative of help-seeking behaviour and the context of beliefs and attitudes towards leprosy. Subsequently we surveyed 356 patients currently receiving treatment for leprosy and recorded specific aspects of each help seeking action and their reports of local beliefs and attitudes towards leprosy. Delay was estimated from time of first symptoms through to start of effective treatment (mean 18 months, median 9 months in Purulia and mean 20 months, median 12 months in Nilphamari). The number of help-seeking actions ranged from 1 to 7. Time committed to first actions contributed 86% (Nilphamari) and 79% (Purulia) to total delay. The most important contributor to delay in the first action occurred when people simply monitored or ignored first symptoms, 80% in Nilphamari and 67% in Purulia. With delay longer than 12 months as outcome, logistic regression analyses identified age over 35 years, multiple visits to practitioners in traditional medicine and multiple visits to health service practitioners as predictive of delay. Attending a nearby clinic and exposure to health education materials were predictive of early presentation reduced delay. PMID- 15881035 TI - Patients' perceptions of reconstructive surgery in leprosy. AB - Reconstructive surgery has made significant advances in correcting deformities in leprosy. However, several patients seem hesitant and unenthusiastic to take advantage of this facility. A study was therefore carried out at the Leprosy Mission Hospital in Kolkata during 1999, to assess patients' perceptions to reconstructive surgery. Of about 300 patients operated during 1991-1997, 117 were interviewed using a Visual Analogue Scale on their pre-operative expectations and satisfaction after surgery. Nearly 40% had their expectations fully met, another 40% partially, and about 10% perceived benefits more than expected. Less than 5% were not satisfied. Results are presented for surgery on hand, foot and eye among males and females, and the implications for future discussed. PMID- 15881036 TI - Impaired contrast sensitivity among leprosy patients with normal visual acuity. AB - Contrast sensitivity is a person's ability to identify an object from its background. Patients with normal visual acuity can have reduced contrast sensitivity and may experience trouble in identifying objects at night or moving around in dimly lit places. Contrast sensitivity has never been studied in leprosy patients having normal visual acuity. This study aimed to determine if contrast sensitivity is impaired in leprosy patients who have normal visual acuity and to identify possible associations with demographic, leprosy and ocular characteristics. A hospital based study measuring and comparing contrast sensitivity using the VCT 6500 chart in 127 consecutive leprosy patients without clinically apparent ocular complications and 123 non-leprosy controls was done. Contrast sensitivity was impaired in leprosy patients in all five spatial frequencies (1.5, 3.0, 6.0, 12 and 18 cycles/degree) investigated. Among leprosy patients, contrast sensitivity falling outside the normative range was associated with increasing age (adjusted OR 1.28, 95% CI: 1.14-1.42), being female (adjusted OR 11.05, 95% CI: 2.93-41.69) and having a grade 2 deformity (adjusted OR 6.43, 95% CI:1.68-24.61). Contrast sensitivity is impaired in leprosy patients having normal visual acuity. Elderly, deformed, female patients are particularly burdened with this vision loss. PMID- 15881037 TI - Progress towards the elimination of leprosy in Nigeria: a review of the role of policy implementation and operational factors. AB - The annual reports of the national leprosy control programme in Nigeria were reviewed to study the trends of the indices of leprosy control from 1992 to 2003 and determine the influence of operational and policy factors. By 2003, both national prevalence and case detection rates had reached below 0.5 per 10,000. Sub-nationally, all except three contiguous States in the Southeast, had prevalence rates below one case per 10,000. Over the 12 years, the prevalence rate decreased by 94-1%, from 7.14 to 0.42 per 10,000, with two periods of rapid decline: 1992-1994 and 1998. Remarkable surges of discharges from multi-drug therapy (MDT) occurred in these same periods. The period 1992-1994 corresponds to the years of introduction of MDT, case reviews, and clean-up of leprosy registers nationwide, while 1998 corresponds to the year the programme adopted the shortened 12-month MDT regime for multibacillary (MB) leprosy. The overall trend of case detection since 1992 was relatively stable, but had three significant periods of initial increase (1992-1994), stability (1994-1999) and recent decline (1999-2003), apparently related to the changing levels of activeness of the national programme. The pattern of new cases detected revealed increasing MB classification and lower disability, but a relatively stable child rate since 1992. The trend of MB proportion was also related to the years of MDT introduction and the adoption of a new leprosy case definition and classification policies. Thus, Nigeria has attained a low leprosy endemic status-mainly through operational and policy influences. The challenges that remain include reducing the relatively high leprosy burden in the Southeastern States and evolving effective case detection interventions that will make an observable impact on the incidence of leprosy. PMID- 15881038 TI - Leprosy control in Myanmar 1952-2003--a success story. PMID- 15881039 TI - Lepromatous lymphadenitis masquerading as lymphoma. AB - A 70-year-old male presented with multiple lymphadenopathy and a strong clinical suspicion of non-Hodgkin's lymphoma. Cervical and axillary nodes were excised and were sent for histopathological evaluation, which revealed aggregates of lepra cells loaded with lepra bacilli. Clinicians practising in leprosy endemic areas should keep lepromatous lymphadenitis in mind while investigating patients with lymphadenopathy. PMID- 15881040 TI - Leprosy with neurofibromatosis--a diagnostic dilemma. AB - The coexistence of leprosy with neurofibromatosis is a rare finding and can pose a diagnostic dilemma. Neurofibromatosis coexisting with borderline tuberculoid leprosy has previously not been reported. We report such a case in a 13-year-old boy where biopsy of clinically uninvolved nerve revealed the presence of acid fast bacilli. A careful diagnostic workup is needed in such cases to ensure proper treatment. Both disorders affect Schwann cells and their relationship merits further consideration. PMID- 15881041 TI - Concerns regarding in vitro and in vivo uses of thalidomide. PMID- 15881042 TI - Syme's/Boyd prosthesis: a new modification. PMID- 15881043 TI - Reversal reaction occurring 16 years after beginning antibacterial treatment- comment. PMID- 15881045 TI - Concept and impact of stigma on discrimination against leprosy sufferers- minimizing the harm. PMID- 15881044 TI - Can we have simpler solutions? PMID- 15881046 TI - Cognitive strategy generation in children with developmental coordination disorder. AB - BACKGROUND: Developmental coordination disorder (DCD) is a condition of impaired quality of movement and occupational performance. It has been hypothesized that the difficulties experienced by children with DCD may in part be due to an impaired ability to use cognitive strategies to solve occupational performance problems. Cognitive Orientation to daily Occupational Performance (CO-OP) is a verbally-based approach to helping children remediate this difficulty. The current pilot study investigated the use of cognitive strategies in children with DCD to determine whether cognitive strategy use is improved by CO-OP. METHODS: Observations of video-recorded sessions of 18 school-aged children were scored for frequency and type of cognitive strategies used. RESULTS: Differences within and between groups revealed changes in the types and frequency of cognitive strategies. CLINICAL IMPLICATIONS: The results of the present study support the use of a cognitively-based approach such as CO-OP in assisting children with DCD in developing cognitive strategies when solving occupational performance problems. However, further research using a larger sample is necessary to fully explore the impact of CO-OP on the strategy use of children with DCD. PMID- 15881047 TI - Understanding the occupational deprivation of refugees: a case study from Kosovo. AB - BACKGROUND: Occupational deprivation as a concept has been presented and discussed in the occupational therapy and occupational science literature for the past several years. The discussion to date, however, has been largely exploratory and theoretical in nature. PURPOSE: This article represents the author's attempt to further understandings of occupational deprivation as a lived experience through research undertaken with Kosovar refugees living in Australia. METHOD AND RESULT: The article focuses on the story of one of the participants of the study which is presented as a case study to illuminate aspects of occupational deprivation as a process occurring over time. The case study is interwoven with interpretive commentary, which serves to highlight key issues as to how occupational deprivation may be experienced by groups of people in different cultural, historic and societal contexts and how it may therefore be addressed by occupational therapists. PRACTICE IMPLICATIONS: The article concludes with reflections on future directions with respect to both further research and professional action. In particular, the focus of professional action is oriented to a population-based approach. PMID- 15881048 TI - The impact of reacher length on electromyography activity and task. AB - BACKGROUND: Reachers are common assistive devices prescribed to help individuals carry out their day-to day activities. PURPOSE: The purpose of this study was to examine the effect of muscle activity in the arm and forearm when using two commercially available reachers of different lengths. METHOD: Fifteen women between the ages of 23 and 25 performed a reaching activity that simulated moving a soup can from a cupboard to a counter. Surface electromyography was used to measure the muscle activity in the arm and forearm and to gain an understanding of the relative contributions of different muscles when using the short-handled reach in comparison to the long-handled reacher. RESULT: The results showed no significant differences between the short and the long reacher conditions for any of the muscles investigated. PRACTICE IMPLICATIONS. The length of a reacher does not necessarily affect the amount of muscle strength required in some muscles and may not be a major factor in the therapist's choice of reacher length. PMID- 15881049 TI - Reflections on ... the recovery paradigm: should occupational therapists be interested? AB - BACKGROUND The mental health literature attests to a paradigm shift from an emphasis on treatment towards a focus on fostering recovery. This paradigm emphasizes many constructs familiar to occupational therapists, including quality of life, empowerment, hope, meaningful activities and work. Occupational therapists are not only familiar with these terms, but have addressed recovery related constructs in both research and practice. METHOD AND PURPOSE: Literature on the recovery paradigm, mental health and occupational therapy was reviewed to gain an understanding of this paradigm and its relationship to occupational therapy in mental health. RESULT: Core occupational therapy beliefs and assumptions are strikingly similar to those purported to be important to fostering recovery. IMPLICATIONS: Occupational therapists should consider taking a keen interest in the construct of recovery. By virtue of professional beliefs and assumptions, occupational therapists are uniquely positioned to assume a leadership role in the area of recovery-related research and practice. PMID- 15881050 TI - Reflection as a means to foster client-centred practice. AB - BACKGROUND: Client-centred practice embraces important concepts about partnership and justice that promote client participation in meaningful occupations. It is important to understand how occupational therapists can engage in and continue to develop client-centred practice within the constraints of everyday settings. PURPOSE AND METHODS: An action research study was undertaken by a group of four occupational therapists to describe insights gained, and to show the effectiveness of participation in reflective education to help foster client centred practice. RESULTS: The six main insights generated by the group involve: congruence, understanding the potential, environmental factors, personal factors, expectations, and lifelong learning. PRACTICE IMPLICATIONS: Although actual change to daily practice was limited, group members felt the reflective group discussion provided them with a launch pad to start a journey toward more client centred practice. This may be the key to moving beyond simply changing the face and language of occupational therapy practice toward a deeper understanding and implementation of client-centred practice to improve client outcomes. Suggestions are provided for implementing reflection to help continue to develop your own practice. PMID- 15881051 TI - Understanding the experience of HIV/AIDS for women: implications for occupational therapists. AB - BACKGROUND: Within the past few years, HIV/AIDS has shifted from being an acute, palliative disease to one that is more chronic and episodic in nature. This shift has major implications for the role of occupational therapy in women's lives. Very few studies, however, have examined the perspective of women living with HIV/AIDS from an occupational therapy perspective. PURPOSE: This qualitative study was designed to examine the experiences of five women living with HIV/AIDS in Southern Ontario and to begin to explore the implications of these findings for occupational therapy. METHOD: Through the implementation of five in-depth interviews, a phenomenological approach was used to explore the lived experience of women with HIV/AIDS. RESULTS: Four main themes emerged: fearing disclosure, experiencing challenges (physical and psychological), having supportive networks, and coping positively with being HIV positive (spirituality and opportunity for living and learning). PRACTICE IMPLICATIONS: There are several potential roles for occupational therapy in working with women who are living with HIV/AIDS More studies need to be pursued in this area of rehabilitation. PMID- 15881052 TI - Meeting the responsibility that comes with the privilege: introducing a taxonomic code for understanding occupation. PMID- 15881053 TI - Fundamental characteristics of a synthesized light source for optical coherence tomography. AB - We describe the fundamental characteristics of a synthesized light source (SLS) consisting of two low-coherence light sources to enhance the spatial resolution for optical coherence tomography (OCT). The axial resolution of OCT is given by half the coherence length of the light source. We fabricated a SLS with a coherence length of 2.3 microm and a side-lobe intensity of 45% with an intensity ratio of LED1:LED2 = 1:0.5 by combining two light sources, LED1, with a central wavelength of 691 nm and a spectral bandwidth of 99 nm, and LED2, with a central wavelength of 882 nm and a spectral bandwidth of 76 nm. The coherence length of 2.3 microm was 56% of the shorter coherence length in the two LEDs, which indicates that the axial resolution is 1.2 microm. The lateral resolution was measured at less than 4.4 microm by use of the phase-shift method and with a test pattern as a sample. The measured rough surfaces of a coin are illustrated and discussed. PMID- 15881054 TI - Establishing a high-accuracy spectral response scale in the near infrared with digital filters. AB - Spectrally invariant detectors are commonly used to interpolate or extrapolate the responsivity of InGaAs detectors in the infrared from absolute calibrations at a few wavelengths. The random noise in such detectors limits the accuracy that can be achieved in a narrowband, double-monochromator setup. We propose the application of a dedicated digital filter, which reduces the uncertainty by 30%, and combine it by calibrating a group of three detectors. The uncertainties are propagated from the observed variance in the relative measurement to the combined uncertainty of 0.4% (2sigma) in the responsivity values of the InGaAs detectors in the range of 1010-1640 nm. PMID- 15881055 TI - Depolarization index and the average degree of polarization. AB - Two single number metrics for depolarization of samples are contrasted: (1) the average degree of polarization of the exiting light averaged over the Poincare sphere and (2) the depolarization index of Gill and Berbenau [Opt. Acta 32, 259 261 (1985); 33, 185-189 (1986)1. The depolarization index is a geometric measure that varies from 0 for the ideal depolarizer to 1 for nondepolarizing Mueller matrices. The average degree of polarization also varies from 0 to 1 and characterizes the typical level of depolarization. Although the depolarization index is very often close to the average degree of polarization, these two metrics can differ by more than 0.5 for certain Mueller matrices. PMID- 15881056 TI - Test of photonic crystal fiber in broadband interferometry. AB - Photonic crystal fibers (PCFs) are microstructured waveguides that are used in metrology, nonlinear optics, and coherent tomography. PCF studies are focused mainly on the improvement of dispersion properties and wide spectral single-mode operating domains. Consequently, in the astronomical context this kind of fiber is a good candidate for use in the design of a fiber-linked version of a stellar interferometer for aperture synthesis. We discuss the potential of these fibers to take advantage of wide spectral single-mode operation. We propose an experimental setup that acts as a two-beam interferometer that uses PCFs to measure fringe contrast at four wavelengths (670, 980, 1328, and 1543 nm), which correspond to the R, I, J, and H astronomical bands, respectively, with the same couple of PCFs. For this purpose we use, for the first time to our knowledge, a piezoelectric PCF optical path modulator. PMID- 15881057 TI - Monochromatic heterodyne fiber-optic profile sensor for spatially resolved velocity measurements with frequency division multiplexing. AB - Investigating shear flows is important in technical applications as well as in fundamental research. Velocity measurements with high spatial resolution are necessary. Laser Doppler anemometry allows nonintrusive precise measurements, but the spatial resolution is limited by the size of the measurement volume to approximately 50 microm. A new laser Doppler profile sensor is proposed, enabling determination of the velocity profile inside the measurement volume. Two fringe systems with contrary fringe spacing gradients are generated to determine the position as well as the velocity of passing tracer particles. Physically discriminating between the two measuring channels is done by a frequency-division multiplexing technique with acousto-optic modulators. A frequency-doubled Nd:YAG laser and a fiber-optic measuring head were employed, resulting in a portable and flexible sensor. In the center of the measurement volume of approximately 1-mm length, a spatial resolution of approximately 5 microm was obtained. Spatially resolved measurements of the Blasius velocity profile are presented. Small velocities as low as 3 cm/s are measured. The sensor is applied in a wind tunnel to determine the wall shear stress of a boundary layer flow. All measurement results show good agreement with the theoretical prediction. PMID- 15881058 TI - Corrections of size-of-source effect and distance effect in radiometric measurements of radiance. AB - Many optical instruments used in quality control of the optical radiation emission level of several devices are limited by the so-called size-of-source effect (SSE) as well as the distance effect (DE) when we are dealing with very accurate measurements. Different authors have studied the SSE and DE and have proposed experimental methods that provide corrections for them. We describe a general method based on the partial coherence theory that allows us to describe and calculate the SSE and DE in any radiometric system with circular apertures. We show some experimental results that verify our proposal. Additionally, as a practical example, we present the corresponding DE and SSE correction factors for a particular geometry. PMID- 15881059 TI - Crossed source-detector geometry for a novel spray diagnostic: Monte Carlo simulation and analytical results. AB - Sprays and other industrially relevant turbid media can be quantitatively characterized by light scattering. However, current optical diagnostic techniques generate errors in the intermediate scattering regime where the average number of light scattering is too great for the single scattering to be assumed, but too few for the diffusion approximation to be applied. Within this transitional single-to-multiple scattering regime, we consider a novel crossed source-detector geometry that allows the intensity of single scattering to be measured separately from the higher scattering orders. We verify Monte Carlo calculations that include the imperfections of the experiment against analytical results. We show quantitatively the influence of the detector numerical aperture and the angle between the source and the detector on the relative intensity of the scattering orders in the intermediate single-to-multiple scattering regime. Monte Carlo and analytical calculations of double light-scattering intensity are made with small particles that exhibit isotropic scattering. The agreement between Monte Carlo and analytical techniques validates use of the Monte Carlo approach in the intermediate scattering regime. Monte Carlo calculations are then performed for typical parameters of sprays and aerosols with anisotropic (Mie) scattering in the intermediate single-to-multiple scattering regime. PMID- 15881060 TI - Virtually calibrated projection moire interferometry. AB - Projection moire interferometry (PMI) is an out-of-plane displacement measurement technique that consists of differencing reference and deformed images of a grid pattern projected onto the test object. In conventional PMI, a tedious process of computing the fringe sensitivity coefficient (FSC), which requires moving the test object or the reference plane to known displacements, is used. We present a new technique for computing the FSC values that is called virtually calibrated projection moire interferometry (VCPMI). VCPMI is based on computer simulations of the conventional PMI process and does not require moving the actual test object or reference plane. We validate the VCPMI approach by comparing results for a flat plate and an airfoil with those made by use of other measurement methods. PMID- 15881061 TI - Surface profiling of a transparent object by use of phase-shifting Talbot interferometry. AB - Talbot interferometry is used to study the surface profile of a transparent object. Periodic patterns are produced by illuminating a grating with a collimated laser beam. The object is placed on the self-image plane of the grating. The deformed grating image, which interferes with another grating, results in the Talbot interferometric fringes. The fringe pattern is recorded on a CCD camera for subsequent analysis, and the phase variation is achieved by a linear translation stage. In this application two specimens are tested to demonstrate the validity of the method; one is a transparent object with a spherical shape with a height of less than 350 microm, and the other is a transparent object with an uneven surface of 50-microm average height. The experimental results are compared with the test results obtained with the mechanical stylus method. PMID- 15881062 TI - Two regions of mode selection in resonators with biprismlike elements. AB - A resonator structure in which one reflector is replaced by a biprismlike reflecting surface is investigated theoretically. It is shown that such a modification leads to two regions of parameters, each with different regimes of mode selection. The first region has an improved laser power output because of the nearly flat-top mode shape. In the second region the biprism is inverted, with the result that the main oscillating mode can be the first odd mode. The line singularity contained in such a mode is one example of singular beams that are employed in various fields, such as micromanipulators and advanced high resolution metrology. PMID- 15881063 TI - Second-order hot image from a scatterer in high-power laser systems. AB - A theory is developed for predicting a second-order hot-image formation in high power laser systems. Light diffracted from a small optical scatterer interferes with an intense original wave in the nonlinear medium to produce a hologram like a Fresnel-zone plate. The theoretical model shows that the hologram produces a negative first-order diffractive wave focused to the traditional hot image and negative second-order diffraction that causes another intense image, namely, a second-order hot image. It is found by analysis that the location of the second order hot image arises in a downstream plane with a half-distance from the medium to the scatterer. Results of the numerical calculations show that the peak intensity of the nonlinear image may reach a level high enough to damage optical components with the increase of the breakup integral (B integral), indicating that the image may also potentially damage expensive optical components in high power laser systems. PMID- 15881064 TI - Scanning near-field optical microscopy as a tool for the characterization of multimode interference devices. AB - We report the scanning near-field optical microscopy (SNOM) characterization of a 4 x 4 multimode interference (MMI) device working at a wavelength of 1.55 microm and designed for astronomical signal recombination. A comprehensive analysis of the mapped propagating field is presented. We compare SNOM measurements with beam propagation-method simulations and thus are able to determine the MMI structure's refractive-index contrast and show that the measured value is higher than the expected value. Further investigation allows us to demonstrate that good care must be taken with the refractive-index profile used in simulation when one deals with low-index contrast structures. We show evidence that a step-index contrast is not suitable for adequate simulation of our structure and present a model that permits good agreement between measured and simulated propagating fields. PMID- 15881065 TI - Engineering the nonlinear phase shift with multistage autoregressive moving average optical filters. AB - We propose and demonstrate the application of concepts from digital filter design in order to optimize artificial optical resonant structures to produce a nearly ideal nonlinear phase shift response. Multistage autoregressive moving average (ARMA) optical filters (ring-resonator-based Mach-Zehnder interferometer lattices) are designed and studied. The filter group delay is used as a measure instead of finesse or quality factor to study the nonlinear sensitivity for multiple resonances. The nonlinearity of a four-stage ARMA filter is 17 times higher than that of the intrinsic material of the same group delay. We demonstrate that the nonlinear sensitivity can be increased within constant bandwidth by allocating more in-band phase or by using higher-order filter structures and that the nonlinear sensitivity enhancement improves with increasing group delay. We also investigate methods to precompensate the nonlinear response to reduce the occurrence of optical bistabilities. The effect of optical loss, including linear absorption and two-photon absorption, is discussed in postanalysis. In addition, we discuss how the improvement in nonlinear response scales with respect to various filter parameters. PMID- 15881066 TI - Collinear holography. AB - A novel reading and writing technology for a holographic storage system called collinear holography is developed. With this method, two-dimensional page data can be recorded as volumetric holograms generated by a reference beam and a signal beam that are bundled on the same axis and irradiated on the recording medium through a single objective lens. The multiplex recording and reconstructing process is demonstrated, and it is presented that optical configuration and the dichroic media disk structure are suitable for a compact system. This method enables us to construct a small volumetric optical disk storage system with CD and DVD upper compatibility. PMID- 15881067 TI - Creation of line defects in holographic photonic crystals by a double-exposure thresholding method. AB - Recording of periodic variations of amplitude and phase by the interference of coherent laser beams in a hologram offers a natural means for creating one-, two , and three-dimensional photonic crystals. For device applications such as waveguides in optical communications, one usually needs to create defects in photonic crystals. We present an analysis and an experimental demonstration of a double-exposure method for creating photonic crystals with line defects. The idea is based on the principle of superposition of holographic grating patterns of different spatial periods while the recording medium is held stationary and on the application of a threshold to the recording medium. We use the same symmetrical optical architecture to achieve nondefective and defective holographic photonic crystals. The technique may be extended to the creation of defects based on functional synthesis by means of Fourier series, by use of light sources of other wavelengths with an appropriate high-contrast recording material. PMID- 15881068 TI - Bulk acousto-optic wavelength agile filter module for a wavelength-multiplexed optical scanner. AB - An acousto-optic tunable filter-based wavelength-selection module with features optimized for a wavelength-multiplexed optical scanner (W-MOS) is proposed and demonstrated. The W-MOS produces high-speed multiple scan beams if it is engaged with an agile tunable source with multiwavelength generation capability. In particular, the proposed fiber-connected module features high-speed, low-loss, narrow-linewidth, and single-multiple wavelength selection by means of radio frequency drive signal control for single- or multiple-beam scan operations. The unique module offers input laser beam power control that in turn delivers the desired scanned laser beam power shaping. Experimental results match module design theory and demonstrate a fast 5.4-micros wavelength selection speed, a low (1.53-dB) fiber-to-fiber optical insertion loss, a 5.55-nm 3-dB spectral width, and a 1500-1600-nm agile wavelength operational band. PMID- 15881069 TI - Phase-only encryption and watermarking based on phase-shifting interferometry. AB - Generally, the reconstruction of an object image from its diffraction field requires both the amplitude and the phase information of this field. We systematically investigated the effects of using only the real part, the imaginary part, or the phase information of the diffraction field to reconstruct the original image for both the binary and the gray-level images. We show that the phase information can yield a better result of image retrieval than the real or imaginary part and that the recovered image from the phase information is satisfactory especially for binary input. On the basis of this idea, a new technique of image encryption and watermarking by use of only one delivered image the phase map of the diffraction field of the original image-through double random-phase encoding is proposed and verified by computer simulations with phase shifting interferometry. This method can greatly cut down the communication load and is suitable for Internet transmission. PMID- 15881070 TI - Normalized differential detection by use of smart pixels with smart illumination. AB - Smart pixels permit rapid signal processing through the use of integrated photodetectors and processing electronics on a single semiconductor chip. Smart pixels with smart illumination can increase the dynamic range and functionality of smart pixels by employing optoelectronic feedback to control the illumination of a scene. This combination of smart pixels and optoelectronic feedback leads to many potential sensor applications, including normalized differential detection, which is modeled and demonstrated here. PMID- 15881071 TI - Enhanced solar irradiance across the atmosphere-sea ice interface: a quantitative numerical study. AB - A radiative transfer model for the coupled atmosphere-sea ice system is used to study the change in downward irradiance across the air-ice interface. Computations demonstrate that the downward solar irradiance can be significantly enhanced across the air-ice interface. The enhancement is mainly due to light in the ice that is scattered upward and then totally reflected by the air-ice interface. It depends primarily on the change in the index of refraction across this interface and the optical properties of the sea ice, but also on the direct solar and sky illumination of the interface. PMID- 15881072 TI - Large-scale wave-front reconstruction for adaptive optics systems by use of a recursive filtering algorithm. AB - We propose a new recursive filtering algorithm for wave-front reconstruction in a large-scale adaptive optics system. An embedding step is used in this recursive filtering algorithm to permit fast methods to be used for wave-front reconstruction on an annular aperture. This embedding step can be used alone with a direct residual error updating procedure or used with the preconditioned conjugate-gradient method as a preconditioning step. We derive the Hudgin and Fried filters for spectral-domain filtering, using the eigenvalue decomposition method. Using Monte Carlo simulations, we compare the performance of discrete Fourier transform domain filtering, discrete cosine transform domain filtering, multigrid, and alternative-direction-implicit methods in the embedding step of the recursive filtering algorithm. We also simulate the performance of this recursive filtering in a closed-loop adaptive optics system. PMID- 15881073 TI - Improvements of the aerosol algorithm in ozone lidar data processing by use of evolutionary strategies. AB - The differential absorption lidar (DIAL) at the Institut fur Meteorologie und Klimaforschung has been upgraded for precise ozone and aerosol studies in the entire troposphere and the lower stratosphere. Its excellent technical performance offers the opportunity to apply improved data processing. The existing inversion algorithm is extended to derive the optical coefficients from the backscatter profiles for three wavelengths. Correlating the correction terms of the DIAL equation and the ozone concentration yields the wavelength dependence of the backscatter and extinction coefficients of the aerosol. Under some conditions, in particular if homogeneous layers are present, the backscatter-to extinction ratio and the reference value can also be retrieved. We find the solutions by applying evolutionary strategies. From the optical coefficients obtained in this way the ozone concentration can be calculated with substantially reduced error. PMID- 15881074 TI - Experimental and theoretical investigations of birefringent holey fibers with a triple defect. AB - We have manufactured and characterized a birefringent holey fiber of a new construction. The birefringence in this fiber is induced by the highly elliptical shape of the core, which consists of a triple defect in a hexagonal structure. Using a hybrid edge-nodal finite-element method, we calculated the spectral dependence of phase and group modal birefringence for spatial modes E11 and E21 in idealized and in real fiber, whose geometry we determined by using a scanning electron microscope. Results of our calculations show that technological imperfections significantly affect the fiber's birefringence. Normalized cutoff wavelengths for higher-order modes relative to the filling factor were also determined for the idealized structure. We observed a significant disagreement between theoretical and experimental values of cutoff wavelengths, which was attributed to high confinement losses near the cutoff condition. We also measured the spectral dependence of the phase and the group modal birefringence for spatial modes E11 and E21. The measured parameters showed good agreement with the results of modeling. PMID- 15881075 TI - Tight packaging of erbium-doped waveguide amplifiers. AB - A numerical model for erbium-doped waveguide amplifiers (EDWAs) containing bent waveguides was developed. The model uses rate-propagation equations and takes into account bend-induced losses in the waveguide with a varying radius of curvature. It is applied to the design of tightly packed erbium-doped waveguide amplifiers under two constraints: minimization of the area required to obtain a predetermined gain and maximization of the gain available from a given area. Numerical calculations based on realistic waveguide parameters demonstrate the possibility of fabricating high-gain, small-size EDWAs. Simple design rules, based on geometric formulas and consideration of bend losses, are given and compared with the results of numerical calculations. PMID- 15881076 TI - Axial optical trapping forces on two particles trapped simultaneously by optical tweezers. AB - Optical tweezers, which utilize radiation pressure to control and manipulate microscopic particles, are used for a large number of applications in biology and colloid science. In most applications a single optical tweezers is used to control one single particle. However, two or more particles can be trapped simultaneously. Although this characteristic has been used in applications, no theoretical analysis of the trapping force or the status of the trapped particles is available to our knowledge. We present our calculation, using a ray optics model, of the axial trapping forces on two rigid particles trapped in optical tweezers. The spherical aberration that results from a mismatch of the refractive indices of oil and water is also considered. The results show that the forces exerted by the optical tweezers on the two particles will cause the two particles to touch each other, and the two particles can be stably trapped at a joint equilibrium point. We also discuss the stability of axial trapping. The calculation will be useful in applications of optical tweezers to trap multiple particles. PMID- 15881077 TI - Thermal diffusivity, specific heat, thermal conductivity, coefficient of thermal expansion, and refractive-index change with temperature in AgGaSe2. AB - Measurement of the key thermo-optic properties of AgGaSe2 in the temperature range below 300 K is reported. Values of these properties on cooling become favorable for the higher average-power operation of nonlinear optical frequency converters using this material. PMID- 15881078 TI - A validated gas chromatographic method for the evaluation of enzymatic enantioselectivity in kinetic resolution applications. AB - An enantioselective gas chromatography (GC) method has been developed and validated for determination of the enantiomers of citronellol in kinetic resolution experiments. S-(-)-beta-Citronellol is a precursor of rose oxide. After solid-phase extraction (SPE) with ethyl acetate, the enantiomers of R-(+) beta-citronellol and S-(-)-beta-citronellol and their corresponding acetate- and butyrate esters were separated through enantioselective GC respectively. The method was validated and found to be reproducible, specific, accurate, and precise. Analyte recoveries and detection limits were also determined. The applicability of this method was shown in a kinetic resolution experiment using lipase A of Bacillus subtilis. PMID- 15881079 TI - Solvation enthalpies and heat capacities of n-alkanes in four polymer phases by capillary gas chromatography. AB - Molar solvation enthalpy (deltasol H(o)298) and molar heat capacity changes (deltasol C(o)p) were determined by gas chromatography for the C6-C12 n-alkanes on four preferred stationary phases (100% polydimethyl siloxane, 50% diphenyl-50% dimethyl polysiloxane, 50% trifluoropropyl methylsiloxane, and polyethylene glycol) in commercial FSOT. Statistical evaluation indicated the temperature independence of deltasol C(o)p in the range 303-393 K. Deltasol H(o)298 depends linearly on the number of carbon atoms in the n-alkanes, but no linearity could be established for deltasol C(o)p of higher homologues on polar columns, which may be due to a more ordered state on the liquid phase. The homologues for which a linear temperature dependence exists demonstrated that deltasol C(o)p is related linearly to the van der Waals volume and the temperature derivative of the density of the stationary phase. The results are consistent with a simple physical explanation at the molecular level. PMID- 15881080 TI - Rapid identification of pathogenic bacteria by capillary electrophoretic analysis of rRNA genes. AB - Molecular diagnosis is playing an increasingly important role in the rapid detection and identification of pathogenic organisms in clinical samples. The genetic variation of ribosomal genes in bacteria offers an alternative to culturing for the detection and identification of these organisms. Here 16S rRNA and 16S-23S rRNA spacer region genes were chosen as the amplified targets for single-strand conformation polymorphism (SSCP) and restriction fragment length polymorphism (RFLP) capillary electrophoresis analysis and bacterial identification. The multiple fluorescence based SSCP method for the 16S rRNA gene and the RFLP method for the 16S-23S rRNA spacer region gene were developed and applied to the identification of pathogenic bacteria in clinical samples, in which home-made short-chained linear polyacrylamide (LPA) was used as a sieving matrix; a higher sieving capability and shorter analysis time were achieved than with a commercial sieving matrix because of the simplified template preparation procedure. A set of 270 pathogenic bacteria representing 34 species in 14 genera were analyzed, and a total of 34 unique SSCP patterns representing 34 different pathogenic bacterial species were determined. Based on the use of machine code to represent peak patterns developed in this paper, the identification of bacterial species becomes much easier. PMID- 15881081 TI - Simultaneous determination of metal ions, amino acids, and other small biogenic molecules in human serum by capillary zone electrophoresis with transient isotachophoretic preconcentration. AB - CE with indirect UV detection was used for the simultaneous determination of lithium, magnesium, calcium, creatinine, carnitine, and a number of amino acids in human serum. The target analytes, positively charged under acidic electrolyte conditions, were separated with positive separation voltage polarity using 10 mM 4-methylbenzylamine, 4.5 mM citric acid, 25% (v/v) methanol at pH 4.05 as background electrolyte providing optimal separation. When analyzing real samples, however, some peaks were broadened due to essentially destacking conditions. In order to maintain the separation efficiency and also enhance the detection sensitivity, transient isotachophoresis (tITP) sample stacking was applied and yielded theoretical plate numbers in the range from 160,000 (arginine) to 350,000 (creatinine). The limit of detection values with tITP preconcentration were 0.11 0.26 mg L(-1) for metal cations, 1.0 mg L(-1) for creatinine, and 1.3-3.9 mg L( 1) for histidine, lysine, arginine, and ornithine. The method precision for peak areas was from 0.4 to 5.0% relative standard deviation using the matrix sodium as internal standard. The accuracy of the developed tITP-CZE system was verified by consistent results for Li+, Mg2+, Ca2+, and creatinine obtained on analyzing two serum certified reference materials. The only sample preparation required was ultrafiltration and acidification (to release protein-bound alkaline earths), and working ranges for individual analytes corresponded well to clinical concentration ranges. PMID- 15881082 TI - Chiral separation of the four stereoisomers of a novel antianginal agent using a dual cyclodextrin system in capillary electrophoresis. AB - Reported here is an analytical method enabling the stereochemical resolution of a new antianginal compound possessing two stereogenic centers, leading to four stereoisomers. Only one of these isomers is currently under development as a novel antianginal agent and consequently, the other three isomers are considered as unwanted chiral impurities. Therefore, an enantioselective method is required in order to check its enantiomeric purity. This paper presents a method exploiting the high efficiency of capillary electrophoresis and the complexing properties of cyclodextrins to achieve the separation of the four stereoisomers of this weakly basic compound (pKa = 7.4). For this purpose, the combination of a neutral cyclodextrin, hydroxypropyl-gamma-cyclodextrin (HP-gamma-CD), and an anionic cyclodextrin, carboxymethyl-beta-cyclodextrin (CM-beta-CD), was added to the separation buffer running in an uncoated silica capillary. After selection of the suitable cyclodextrin system, satisfactorily separation conditions were as follows: 30 mM phosphate buffer (pH 6.4) containing 10 mM of HP-gamma-CD and 10 mM of CM-beta-CD, running voltage +30 kV. The resulting run time and resolutions were respectively about 17 min and between 1.95 and 2.84. Linearity curves (0.993 < r2 < 0.998) are also shown. PMID- 15881083 TI - Simultaneous separation and determination of Tarabine PFS and Adriblastine using micellar electrokinetic chromatography and high performance liquid chromatography. Application to some biological fluids. AB - The simultaneous determination of Tarabine PFS and Adriblastine by two independent techniques, viz. micellar electrokinetic chromatography (MEKC) and high performance liquid chromatography (HPLC), has been studied. For MEKC analysis, separations and identifications were accomplished using uncoated fused silica capillaries and injections were performed in the hydrodynamic mode. The running buffer consisted of 0.05 M borate/phosphate pH 8.70, with 0.10 M SDS at an operating voltage of 15.0 kV and the temperature held at 25.0 degrees C. Under these conditions, the migration times of Tarabine PFS and Adriblastine were 2.70 and 6.40 min, respectively. Calibration curves were established for 0.010-0.300 microg/mL (r = 0.99) Tarabine PFS and 8.000-120.0 microg/mL (r = 0.99) Adriblastine. The limit of detection (LOD) was estimated and found to be 0.003 and 3.000 microg/mL of Tarabine PFS and Adriblastine, respectively. The limit of quantitation (LOQ) was found to be 0.009 and 8.000 microg/mL of Tarabine PFS and Adriblastine, respectively. For HPLC analysis, separations and determinations were performed on teicoplanin stationary phase with reversed mobile phase containing methanol:buffer pH 4.05 (20.0:80.0%, v/v) at 285 nm. Calibration curves were established for 3.000-90.00 microg/mL (r = 0.99) Tarabine PFS and for 10.00-120.0 microg/mL (r = 0.99) Adriblastine. LOD and LOQ were estimated and found to be 0.950 and 2.050 microg/mL of Tarabine PFS and 3.130 and 9.250 microg/mL of Adriblastine, respectively. Both MEKC and HPLC methods were applied for the simultaneous determination of analytes in urine samples. It was found that 8.00-10.0% (Tarabine PFS) and 13.0-15.0% (Adriblastine) of the injected dose was recovered in urine samples with 99.5-102% recovery. PMID- 15881084 TI - Micellar electrokinetic chromatography method for the determination of sulfamethoxazole, trimethoprim and their main metabolites in human serum. AB - A complete analytical procedure, including sample clean-up and a micellar electrokinetic chromatographic method, is presented for the determination of sulfamethoxazole, trimethoprim, and their main metabolites by using 20 mmol L(-1) borate buffer (pH 9.3), 25 mmol L(-1) sodium dodecylsulfate, and 5% v/v acetonitrile as electrolyte. The separation was carried out at 30 kV and 20 degrees C in a fused silica capillary (60.2 cm x 75 microm inner diameter) fitted with a window in the capillary cartridge of 100 x 800 microm. The detector response was linear from the limit of quantification to 3 mg L(-1) for the individual components. The limits of quantification ranged from 0.13 up to 0.24 mg L(-1). The method was applied to human serum, previously spiked at different concentrations of all the analytes, and recoveries between 95% and 108% were obtained. PMID- 15881085 TI - Extended thermodynamic approach to ion interaction chromatography. Influence of the chain length of the solute ion: a chromatographic method for the determination of ion-pairing constants. AB - The influence of the chain length in Ion Interaction Chromatography (IIC), previously named Ion Pair Chromatography, was investigated. The system examined is a C18 stationary phase and a variable, salt-controlled, mixture of a phosphate buffer pH 2.1 in methanol containing tetrabutylammonium bromide as Ion Interaction Reagent. IIC proves to be a good alternative to the well established spectrophotometric and conductometric methods of obtaining thermodynamic ion pairing constants. The latter increase with increasing analyte chain length and decrease with increasing methanol concentration in the eluent: both results support a physical chemical description of the hydrophobic ion-pairing process at variance with the classical, Bjerrum-type modelling of the purely electrostatic interaction between inorganic ions. The calculated deltaG(o) are reliable because they compare to non-chromatographic estimates of deltaG(o) for similar systems. The present extended thermodynamic approach is able to predict and explain why the purely electrostatic approach works better for lower analyte lipophilicity or higher organic modifier concentration. PMID- 15881086 TI - Determination of fluoroquinolones in edible animal tissue samples by high performance liquid chromatography after solid phase extraction. AB - In the present work, a rapid, accurate, and sensitive method has been developed for the quantitative determination of five fluoroquinolones (enoxacin, ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin) in edible animal tissues (muscle tissue, liver, kidney, and eggs). The separation was accomplished on an Inertsil (250 x 4 mm) C8, 5 microm, analytical column, at ambient temperature within 15 min. The mobile phase consisted of a mixture of citric acid (0.4 mol L(-1))-CH3OH CH3CN (87:9:4% v/v). UV detection at 275 nm yielded the following limits of detection: 100 pg per 20 microL injected volume for enoxacin, norfloxacin, and ciprofloxacin, 20 pg for ofloxacin, and 200 pg for enrofloxacin. Peaks in real samples were identified by means of a photodiode array detector. The method was validated in terms of intra-day (n = 8) and inter-day (n = 8) precision and accuracy. Tissue samples were purified from endogenous interference by solid phase extraction using Oasis HLB cartridges. The solid-phase extraction protocol was optimized in terms of retention and elution. Recovery rates at fortification levels of 40, 60, and 80 ng/g ranged from 82.5% to 111.1%. The applicability of the method was examined using real samples from a chicken treated orally with the five studied fluoroquinolones. PMID- 15881087 TI - Retention process in reversed phase TLC systems with polar bonded stationary phases. AB - The retention of a solute in RP chromatography is a very complex process which depends on many factors. Therefore, the study of the influence of a mobile phase modifier concentration on the retention in different reversed phase chromatographic systems is very important for understanding the rules governing retention and mechanisms of substance separation in a chromatographic process. Composition changes and the nature of mobile phases enable tuning of the separated analytes' retention over a wide range of retention parameters and optimization of the chromatographic process as well. Optimization of the chromatographic process can be achieved by several different methods; one of them is the so-called interpretative strategy. The key approach adopted in this strategy is the implementation of adequate retention models that couple the retention of solute with the composition of a mixed mobile phase. The use of chemically bonded stationary phases composed of partially non-bonded silica matrix and organic ligands bonded to its surface in everyday chromatography practice leads to questions of the correct definition of the retention model and the dominant retention mechanism in such chromatographic systems. The retention model for an accurate prediction of retention factor as a function of modifier concentration and the heterogeneity of the adsorbent surface should be taken into consideration. In this work the influence of mobile-phase composition on the retention of sixteen model substances such as phenols, quinolines, and anilines used as test analytes in different RP-TLC systems with CN-, NH2-, and Diol-silica polar bonded stationary phases has been studied. The aim of this study is to compare the performance of three valuable retention models assumed as the partition, adsorption/partition, and adsorption mechanism of retention. All the models were verified for different RP-TLC systems by three statistical criteria. The results of investigations presented in this work demonstrate that the best agreement between the experimental and calculated Rf values was obtained by the use of new-generation retention models, which assume heterogeneity of adsorbent surface. The results reported here show that heterogeneity of the adsorbent surface may be important in analysis of the elution process in liquid chromatography. Consideration of the goodness of fit for the experimental data to the examined retention models is in conformity with the adsorption mechanism of retention on all polar bonded stationary phases in most eluent systems for most investigated compounds. PMID- 15881088 TI - Development of a HPTLC method for in-process purity testing of pentoxifylline. AB - A HPTLC method for the separation and identification of pentoxifylline and related substances, impurities of reaction partners, and side reaction products has been developed using different mobile and stationary phases. For quantitative assay of possible by-products as impurities, LiChrospher RP-18 F254s chromatoplates, acetone-chloroform-toluene-dioxane (2:2:1:1 v/v) as a mobile phase, and detection at 275 nm were employed. Linearity (r > or = 0.997), recovery (86.5-115.5%), and determination limit (0.1-0.6%) were evaluated and found to be satisfactory. This method enables monitoring of the synthesis, as well as purity control of pentoxifylline-containing raw materials and pharmaceuticals. PMID- 15881089 TI - A rapid densitometric method for simultaneous quantification of gallic acid and ellagic acid in herbal raw materials using HPTLC. AB - Gallic acid and ellagic acid are two widely occurring phenolic compounds of plant origin, to which many biological activities including anticancer and antiviral activity have been attributed. A simple HPTLC method has been developed for the simultaneous quantification of gallic acid and ellagic acid. The method was validated for precision, repeatability, and accuracy. Instrumental precision was found to be 0.083 and 0.78, and the repeatability of the method was found to be 1.07 and 1.50 (% CV) for gallic acid and ellagic acid, respectively. The accuracy of the method was checked by a recovery study conducted at two different levels and the average percentage recovery was found to be 101.02% for gallic acid and 102.42% for ellagic acid. The above method was used for the quantification of gallic acid and ellagic acid content in seeds of Abrus precatorius Linn., whole plant of Phyllanthus maderaspatensis Linn., and flowers of Nymphaea alba Linn. The proposed HPTLC method for the simultaneous quantification of gallic acid and ellagic acid was found to be simple, precise, specific, sensitive, and accurate and can be used for routine quality control of herbal raw materials and for the quantification of these compounds in plant materials. PMID- 15881090 TI - Application of water as a solvent in microwave-assisted extraction for analysis of PCBs and CBzs in fly ash. AB - Polychlorinated biphenyls (PCBs) and chlorobenzenes (CBzs) are two classes of dioxin precursors formed in municipal solid waste incinerators (MSWIs); they produce negative health effects similar to those of dioxins. Reducing the analytical time required for determining the concentrations of these compounds in MSWIs is important for quickly evaluating their importance and assessing associated health risks. In the present study, water is used as a safe and environmentally friendly solvent in microwave-assisted extraction (MAE) for PCB and CBz analyses. MAE is compared with traditional Soxhlet extraction (SE) to determine the extraction efficiencies. The evaluation of extraction efficiencies shows that MAE has a high extraction efficiency compared with that of SE when water content is lower than 60%. Furthermore, the extraction time and organic solvent consumption are reduced with MAE compared with SE. PMID- 15881091 TI - [Dyspnea: pathophysiological and clinical aspects]. PMID- 15881092 TI - [Functional condition of bone tissue in smoking and non-smoking patients with chronic obstructive pulmonary disease]. AB - AIM: To evaluate bone and calcium-phosphorus metabolism, bone tissue density (BTD) in smokers and non-smokers with chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS: The study included 120 patients with COPD; smokers (n=68), smokers in the past (n=8) and non-smokers (n=44). Control 80 healthy subjects were matched by age and sex. Bone metabolism was estimated by concentration of osteocalcin (OC), markers of bone resorption (TRAP, betaCrossLaps-betaCL). RESULTS: Smoking aggravates disturbance of calcium metabolism in COPD leading to hypocalcaemia and hypercalciuria. In smokers with COPD bone remodeling dysfunction is caused by suppression of osteogenesis and enhancement of resorption, in non-smokers--intensification of both resorption and osteogenesis. The analysis of correlations has shown that there is a close correlation between the level of TRAP, bone mineral density scores and indices of pack-years (r = -075, p < 0.01; r = -0.6, p < 0.01, respectively). Anamnesis of smoking has a positive correlation with TRAP (r = 0.85, p < 0.001) and a negative correlation with bone density (r = -0.8, p < 0.01) and OC (r = -0.55, p < 0.01). CONCLUSION: Duration of smoking (total pack-years) is an additional marker of osteopenic syndrome in COPD. This is confirmed by close correlation of this parameter with bone density and markers of bone resorption. PMID- 15881093 TI - [What is the danger of long-term application of beta blockers in patients with ischemic heart disease and concomitant chronic obstructive pulmonary disease?]. AB - AIM: To show specific features of respiratory disoders in patients with ischemic heart disease (IHD) and chronic obstructive pulmonary disease (COPD) taking atenolol for a long time. MATERIAL AND METHODS: A long-term study of external respiration function was made in 287 patients with IHD and COPD treated with berodual and atenolol (group 1) vs 74 matched patients taking berodual only. RESULTS: Atenolol treated patients demonstrated more pronounced respiratory disorders. Small respiratory tracts were involved earlier with appearance of expiratory peripheral collapse and early expiratory closure of the respiratory tracts. Diffuse ability of the pulmonary tissue deteriorated. Conclusion. Atenolol has a negative effect on the respiratory system, so it is necessary to control external respiratory function in the course of treatment. PMID- 15881094 TI - [An information model as a method of mathematical processing of clinical material (chronic bronchitis as illustration)]. AB - AIM: To design information clinical-prognostic formalized model of chronic bronchitis (CB). MATERIAL AND METHODS: The study of clinicofunctional, immunometabolic manifestations in CB remissions in 1135 CB patients and genealogical study of 24 families in three generations (83 persons) were performed. Clinical, functional, biochemical, immunological examinations were made. RESULTS: An algorithm of information model of CB consisting of 5 principal steps and technology of information processing of clinical material are proposed. A data matrix was developed in the form of blocks (passport data, former life style, complaints, disease history, characteristics of objective status, functional, clinical, biochemical, immunological parameters) and information analysis was made using a selected mathematical method (correlation, cluster, factorial, regression analyses). CONCLUSION: The proposed methodological approach allows assessment of interaction of such factors as heredity, environment, life style and their effect on CB risk, clinical picture, course; design of program of primary and secondary prophylaxis and assessment of treatment efficacy. PMID- 15881095 TI - [Pulmonary pathology in patients with systemic lupus erythematosus]. AB - AIM: To characterize pulmonary lesion in systemic lupus erythematosus (SLE) on the basis of clinical device and biochemical examination regarding features of the disease onset and development. MATERIAL AND METHODS: The study included 60 SLE patients. Mean age 41.1 +/- 1.32 years. Mean SLE duration 12.42 +/- 1.06 years. Activity according to the SLEDAI and ECLAM indices--16.23 +/- 0.93 and 3.09 +/- 0.18 scores, respectively. The comparison groups: 30 patients with bronchial asthma (BA), 15--with chronic bronchitis (CB), 15--with chronic obstructive bronchitis (COB), 30 healthy donors. The following parameters were studied: spirometric, bodyplethismographic evidence, diffuse ability of the lungs (DAL), plasm concentrations of adrenaline, noradrenaline, dopamine, serotonine, histamine, hemodynamics, anxiety, depression, social adaptation (quality of life) and vegetative dysfunctions. Statistics were obtained with BIOSTATISTIKA program. RESULTS: DAL depends on duration of SLE, severity of lung hypertension (LH), severity of anemia. LH in SLE deteriorated vegetative disorders and social adaptation. Lowering of plasm dopamine concentration was accompanied with LH, formation of vegetative dysfunction and worse social adaptation. CONCLUSION: Affection of the lungs in SLE patients runs without evident clinical symptoms. Initial signs of lung affection manifest with low DAL, LH, moderate restrictive, obstructive and mixed disorders of external respiration function. PMID- 15881096 TI - [Misdiagnosis of extrahospital pneumonia at the prehospital stage]. PMID- 15881097 TI - [Monitoring and treatment of severe bronchial ashma in adults: results of national multicenter trial NABAT]. AB - AIM: To study quality of monitoring, features of a course and therapy of severe bronchial asthma (BA) in Russia. MATERIALS AND METHODS: The trial recruited 378 patients with severe bronchial asthma (SBA) in 7 cities of Russia. Each center participating in the study filled in individual case sheets recording demographic, clinical, monitoring, pharmacotherapeutic data. A special focus was on factors influencing the disease course. The chi-square and Mann-Witney criteria were used. RESULTS: It was found that current recommendations of BA patients' management are not used in full. Most of the patients do not receive adequate therapy. Phenotypes of uncontrolled course of severe BA are prevalent. The key role of adequate management of severe BA patients in control of the disease is shown. CONCLUSION: Wider use of updated recommendations on therapy and monitoring of BA with special focus on patients with uncontrolled course of the disease is urgent. Fulfilment of all the components of management of a patient with bronchial asthma recommended by GINA including the patient education and control over the disease triggers contribute much to effective treatment and control over BA. PMID- 15881098 TI - [Lifespan of patients with primary pulmonary hypertension]. AB - AIM: To design a mathematical express model of expected lifespan of patients with primary pulmonary hypertension (PPH). MATERIAL AND METHODS: A retrospective analysis of 418 case histories of 132 PPH patients (81 females, 51 males, age 16 57 years). Ninety one of them were hospitalized at least twice. The following characteristics of the patients were registered: a PPH form, age at onset, gender, morphological structure of the lung, date of the first visit to doctor. Relationship of these factors with lifespan was analysed and the express formula of expected lifespan was calculated using the multifactorial analysis of variance with standard computer statistical programs. RESULTS: Lifespan was studied for dependence on static factors free of therapy effects (gender, age, form of PPH). The presence of a factor named "duration of the disease development" (DDD) is suggested. A correlation was found between DDD and lifespan in PPH patients explaining survival by 84.5%. The formula of the prognosis was derived for patients with known morphological structure of the lung in whom the above dependence explains survival by 93.5%. CONCLUSION: Among effect modifiers, of significance for survival are age, PPH form, morphological structure of the lung. An artificial index DDD was used for deriving a formula for prognosis of survival for a newly hospitalized PPH patient. PMID- 15881099 TI - [A case of isolated pulmonary lymphogranulomatosis]. PMID- 15881100 TI - [Social stress as a risk factor for ischemic heart disease]. AB - AIM: To study dynamics of ischemic heart disease in exposure of the population of an industrial city to social-economic stress. MATERIAL AND METHODS: Official annual records of Saratov city Health Administration for 1989-1998. In public health evaluation, cardiovascular morbidity is the index rapidly reacting to social-economic changes. Descrete data of myocardial infarction and angina prevalence for the analysed time were approximated by polynomes of high degree. As a result, the first and second derivatives--speed and acceleration--were obtained. RESULTS: A rise in MI morbidity reflected a rise in social tension. The disturbance of the population system of an industrial center (in conditions of a social stress) is accompanied by marked deformation of the profile of cardiovascular diseases in the industrial region: the growth of MI prevalence is associated with lowering of angina pectoris morbidity. CONCLUSION: A rise in MI incidence rate is proposed as an indicator of social-economic stability or instability. Instability of IHD morbidity in the population allows consideration of social stress as its risk factor. PMID- 15881101 TI - [Body mass loss due to orlistat therapy and multiple factors of cardiovascular risk]. AB - AIM: To study effects of body mass loss due to orlistat on carbohydrate and lipid metabolism, insulin resistance, 24-h profile of arterial pressure (AP), left ventricular myocardial hypertrophy, brain perfusion in patients with metabolic syndrome (MS). MATERIAL AND METHODS: Thirty middle-aged patients with MS entered the trial. They received orlistat in a dose 120 mg twice a day for 24 weeks. Before and after the treatment the patients' carbohydrate and lipid metabolism, insulin resistance were studied, 24-h monitoring of arterial pressure, echo cardiography were made. Brain perfusion was studied with single-photon emission computed tomography in 18 patients. Results. All the patients lost much weight. This was accompanied with improved indices of AP profile, metabolism of carbohydrates and lipids, insulin resistance, left ventricular hypertrophy, brain perfusion. Conclusion. Orlistat treatment weakens basic factors of cardiovascular risk. PMID- 15881102 TI - [Applicability of fozinopril for correction of basic manifestations of metabolic syndrome in women with arterial hypertension]. AB - AIM: To evaluate effects of fozinopril on basic manifestations of metabolic syndrome in women with moderate and severe arterial hypertension (AH). Material and methods. Sixty women with AH aged 45 to 65 years entered the trial. Mean duration of the disease was 11 years. Arterial pressure (AP), carbohydrate, lipid and purin metabolism, lipid peroxidation, erythrocytic membranes resistance were examined before the trial and followed up for 6 months of the treatment. RESULTS: Fozinopril significantly reduced systolic and diastolic AP. A 6-month course of the drug brought target AP in 85.4% examinees, a good response was achieved in 96.4%. The treatment also significantly reduced the levels of insulin, C-peptide, triglycerides, increased content of HDLP cholesterol, relieved insulin resistance. Fozinopril demonstrated antioxidant, membrane stabilizing and hypouricemic efficacy. CONCLUSION: High antihypertensive efficacy and a positive action on basic manifestations of metabolic disorders allow us to recommend fozinopril as a basic drug for treatment of women with moderate and severe AH concurrent to insulin resistance syndrome. PMID- 15881103 TI - [10-year trends in body weight in men and women in Novosibirsk (1985-1995)]. AB - AIM: To assess 10-year trends in prevalence of overweight and obesity in Novosibirsk population. MATERIAL AND METHODS: The data of three population surveys (a total of 9714 men and women aged 25-64 years) carried out according to WHO MONICA program. RESULTS: Obesity prevalence among males in 1985-1989 tended to a small rise without changes during subsequent 5 years. In females the prevalence of obesity and overweight decreased in 1985-1994. CONCLUSION: 10-year trends in prevalence of overweight and obesity were insignificant in men and beneficial in women of Novosibirsk population while the frequency of increased body mass in women remains relatively high. PMID- 15881104 TI - [Arterial hypertension, disorders of carbohydrate and lipid metabolism in obese perimenopausal females]. AB - AIM: To examine prevalence of arterial hypertension (AH), disorders of carbohydrate and lipid metabolism in females with obesity in perimenopause. MATERIAL AND METHODS: One-stage trial enrolled 358 obese females in menopause seeking advice in the city center for health and family planning in Ekaterinburg (mean age 50.5 +/- 5.4 years). RESULTS: 111 (31%) females suffered from obesity, 154 (43%) had overweight, 93--normal body mass. A relative risk to develop AH in overweight was 2.61, in obesity--3.52. Diabetes mellitus was in 15 females with obesity, disturbed glucose tolerance was in 38 females with overweight. CONCLUSION: Body weight loss is an important component of perimenopausal women rehabilitation. PMID- 15881105 TI - [Regulatory-adaptive abilities of hypertensive patients treated with lisinopril and atenolol]. AB - AIM: To examine regulatory-adaptive possibilities of hypertensive patients treated with lisinopril and atenolol. MATERIAL AND METHODS: Monotherapies with lisinopril and atenolol were compared in relation to effect on arterial pressure and parameters of cardiac-respiratory synchronism after a single and 5-week intake in 37 patients. RESULTS: Lisinopril increased in 19 patients the range of synchronization by 28.5% (p < 0.05) after a single dose and by 40% (p < 0.001) after 5-week treatment. Atenolol had an adequate hypotensive effect in 18 patients but promoted a decrease of range sychnonization by 41% (p < 0.00001) after a single intake. CONCLUSION: Hypotensive drugs should be studied for effects on regulatory-adaptive ability of the organism. PMID- 15881106 TI - [Anaphylactic reactions and their therapy]. AB - AIM: To ascertain the causes, clinical picture of severe anaphylactic reactions, early diagnosis, decision making. MATERIAL AND METHODS: Examination covered 156 patients (59 males and 97 females) aged 15-81 treated in the department of critical care of Kaunas Medical University for the last 8 years. RESULTS: AR were considered anaphylactoid in 60.2%, acute allergic of type I in 39.7%. AR was timely diagnosed in 85.2%, only 52.5% patients received adequate treatment. Of 156 patients, 11 (7.05%) died. CONCLUSION: The doctors should be alert in respect to possible AR and render assistance immediately. This allows to save all the patients. PMID- 15881107 TI - [Results of an open prospective controlled randomized comparative trial of flurbiprofen and paracetamol efficacy and tolerance in patients with throat pain]. AB - AIM: To compare efficacy and safety of strepfen (flurbiprofen) to those of oral paracetamol in acute infections of the upper respiratory tracts for relief of throat pain (TP). MATERIAL AND METHODS: Of 60 patients 30 ones took strepfen, 30 controls took paracetamol. The intensity and dynamics of throat pain and pharyngeal discomfort were assessed by 10-point subjective visual-analog scale. RESULTS: Strepfen is similar by an analgetic effect to paracetamol. Strepfen acts rapidly (in 5-15 min) and its analgetic effect lasts about 3 hours. Paracetamol begins to relieve pain 30-45 min after its intake. The drug is well tolerated, serious side effects are absent. CONCLUSION: Strepfen can be recommended for relief of throat pains and pharyngeal paresthesia in patients with acute and chronic pharyngeal pathology. PMID- 15881108 TI - [Resynchronising cardiac therapy in a patient with chronic heart failure without expansion of QRS complex]. PMID- 15881109 TI - [Ischemic renal disease in a female with mesangiocapillary glomerulonephritis with nephrotic syndrome]. PMID- 15881110 TI - [Current possibilities of the treatment of chronic obstructive pulmonary disease: bronchodilating medication of long duration]. PMID- 15881111 TI - [Some mechanisms of pulmonary hypertension development in patients with chronic obstructive pulmonary diseases]. PMID- 15881112 TI - [Insulin resistance and diabetes mellitus due to therapy with glucocorticosteroids]. PMID- 15881113 TI - Variability in the composition of anti-oxidant compounds in Echinacea species by HPLC. AB - A fast and reliable HPLC method for the determination of caffeic acid derivatives (caftaric acid, chlorogenic acid, caffeic acid, cynarin, echinacoside and cichoric acid) in various species of the genus Echinacea has been developed. Extraction of root samples by magnetic stirring with 80% methanol aqueous solution at room temperature allowed the complete recovery of all compounds of interest. Root extracts were analysed on a reversed-phase column with gradient elution and photodiode array detection. Caffeic acid derivatives showed differential qualitative and quantitative distributions in Echinacea species. The total amount of phenolic compounds ranged from 33.95 to 0.32 mg/g. The highest contents of caffeic acid derivatives were found in E. paradoxa var. paradoxa, E. paradoxa var. neglecta and E. purpurea, followed by E. angustifolia var. angustifolia, E. simulata, E. pallida and E. laevigata, whilst E. tennesseensis, E. sanguinea and E. atrorubens had low amounts of phenolic compounds. The radical scavenging activities of methanolic extracts of roots of Echinacea species was evaluated in vitro using the DPPH* radical scavenging method. The EC50 values of the samples ranged from 122 to 1223 microg/mL. The radical scavenging activities of the root extracts were correlated with the content of phenolic compounds, with a correlation coefficient (r2) of 0.923. PMID- 15881114 TI - Isolation and identification of arctiin and arctigenin in leaves of burdock (Arctium lappa L.) by polyamide column chromatography in combination with HPLC ESI/MS. AB - A simple method involving polyamide column chromatography in combination with HPLC-PAD and HPLC-ESI/MS for isolating and identifying two kinds of lignans, arctiin and arctigenin, in the leaves of burdock (Arctium lappa L.) has been established. After extraction of burdock leaves with 80% methanol, the aqueous phase of crude extracts was partitioned between water and chloroform and the aqueous phase was fractionated on a polyamide glass column. The fraction, eluting with 100% methanol, was concentrated and gave a white precipitate at 4 degrees C from which two main compounds were purified by semi-preparative HPLC. In comparison with the UV and ESI-MS spectra and the HPLC retention time of authentic standards, the compounds were determined to be arctiin and arctigenin. The extraction/separation technique was validated using an internal standard method. PMID- 15881115 TI - HPTLC determination of vasicine and vasicinone in Adhatoda vasica. AB - A simple high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous determination of the pharmacologically important quinazoline alkaloids vasicine and vasicinone in Adhatoda vasica. The assay combines the separation and quantification of the analytes on silica gel 60 GF254 HPTLC plates with visualisation under UV and scanning at 270 and 281 nm. Using this technique, the alkaloidal content of different parts of the title plant have been determined. PMID- 15881116 TI - An HPLC-PAD method to analyse flavonoid glycosides and styrylpyrones from Cryptocarya species (Lauraceae). AB - An HPLC-PAD method has been developed in order to evaluate simultaneously the main secondary metabolites, flavonoid glycosides and styrylpyrones, of leaves of Cryptocarya moschata. The sample preparation, consisting of extraction, liquid liquid extraction and centrifugation, requires minimum sample manipulation but produces high yields with reproducibility, selectivity and simplicity. HPLC on a C18 column presents each class of metabolites grouped and with good resolution of the main compounds. The experimental conditions can be used to study inter- and intra-specific variability of secondary metabolites in Cryptocarya spp. PMID- 15881117 TI - CGC-MS of alkaloids in Leucojum aestivum plants and their in vitro cultures. AB - Underivatised alkaloid mixtures extracted from intact plants and in vitro cultures of Leucojum aestivum (Amaryllidaceae) were investigated by capillary GC MS. Excellent peak resolution for the alkaloids was exhibited and isomers of galanthamine and N-formylnorgalanthamine were well separated. Fourteen alkaloids of galanthamine, lycorine and crinane types were identified, 11 in the intact plants and eight in the in vitro cultures. PMID- 15881118 TI - Direct identification and quantitative determination of costunolide and dehydrocostuslactone in the fixed oil of Laurus novocanariensis by 13C-NMR spectroscopy. AB - The fixed oil of Laurus novocanariensis (previously L. azorica) contains mostly glycerides together with minor non-saponifiable compounds. The direct identification and quantitative determination of costunolide and dehydrocostuslactone, two sesquiterpene lactones components of the oil that exhibit biological activities, is described. The analysis was carried out using 13C-NMR spectroscopy (signal acquisition with inverse gated decoupling of protons; diglyme as internal standard) without separation, derivatisation or any sample preparation. PMID- 15881119 TI - Solid-phase extraction and HPLC-MS profiling of pyrrolizidine alkaloids and their N-oxides: a case study of Echium plantagineum. AB - Pyrrolizidine alkaloids and their N-oxides can be extracted from the dried methanolic extracts of plant material using dilute aqueous acid. The subsequent integration of solid-phase extraction (with a strong cation exchanger) of the alkaloids and N-oxides from the aqueous acid solution, together with analysis using HPLC-ESI/MS, provides a method for the simultaneous profiling of the pyrrolizidine alkaloids and their N-oxides in plant samples and the collection of useful structural data as an aid in their identification. The N-oxide character of the analytes may be confirmed by treating analytical samples with a redox resin and observing the formation of the corresponding parent pyrrolizidine alkaloids. The present case study of Echium plantagineum highlighted a higher ratio of N-oxides to the parent tertiary bases than has been previously reported. Furthermore, a higher proportion of acetylated pyrrolizidine-N-oxides was observed in the flower heads relative to the leaves. Six pyrrolizidine alkaloids or pyrrolizidine-N-oxides, not previously reported from E. plantagineum, were tentatively identified on the basis of MS and biogenetic considerations. Three of these, 3'-O-acetylintermedine/lycopsamine, leptanthine-N-oxide and 9-O angelylretronecine-N-oxide, have been reported elsewhere, whilst three others, 3' O-acetylechiumine-N-oxide, echimiplatine-N-oxide and echiuplatine-N-oxide, appear unreported from any other source. PMID- 15881120 TI - Analysis and identification of lignans in Phyllanthus urinaria by HPLC-SPE-NMR. AB - HPLC-SPE-NMR has been applied to the analysis of a number of structurally similar lignans present in Phyllanthus urinaria. A reversed-phase HPLC system, using a C8 column eluted with tetrahydrofuran-water-methanol and detected at 225 nm was developed that gave good resolution for seven lignans from Phyllanthus urinaria. Coupling of this system with SPE-NMR provided clean 1H-NMR spectra for nine lignans present in 4 mg of a lignan-rich fraction, equivalent to ca. 1.0 g of dry plant material. The result demonstrates that HPLC-SPE-NMR is a very efficient and powerful tool for the unambiguous identification of natural products that have been previously characterised. PMID- 15881121 TI - Application of capillary-scale NMR for the structure determination of phytochemicals. AB - Employing a capillary-scale NMR probe enables the miniaturisation of structure determination and de-replication of purified natural products from plants using only 5-100 microg of material. Approximately 5 microg are required to perform one dimensional proton and two-dimensional homonuclear (COSY and NOESY) NMR experiments; some 30 microg are needed to acquire HMQC- or HSQC-NMR spectra; ca. 75-100 microg are necessary to measure HMBC-NMR spectra; and around 200 microg of a compound are needed to perform 13C- and DEPT-NMR experiments. In order to illustrate the integration of the outputs from high-throughput natural product chemistry methods with the capabilities of the state-of-the-art CapNMR technology, the preparation of a natural product library from the extract of Penstemon centranthifolius, and the subsequent isolation, purification and structure determination of six known iridoid glycosides with 25-300 microg of material are presented. PMID- 15881122 TI - [Alternative understanding of formation of ketone bodies, kinetics of beta oxidation of fatty acids and of pathogenesis of ketoacidosis]. PMID- 15881123 TI - [Methodological approaches to the determination of trophoblastic beta1 glycoproteins]. PMID- 15881124 TI - [Quantitation of glucocorticoids in blood serum by high-efficiency liquid chromatography]. AB - The paper presents a method of quantitation of endogenous and exogenous glucocorticoids by high-efficiency liquid chromatography. The developed technique is implemented on the basis of Russian equipment and Russian reagents (except for etalons). A "Milichrom" Russian chromatograph is convenient and safe in operation; as for the detection limit, it is not inferior to its foreign analogues. The suggested technique is simple, safe and can be implemented at city and district patient-care facilities. PMID- 15881125 TI - [Specific features of protease and antiprotease activity of blood in women with external endometriosis]. AB - 32 healthy fertile women (control group) and 94 women with different-severity external endometriosis (EE) were examined. The activity of cathepsin D, plasminogen, AT-III, protein C, acid alpha1-antitrypsin as well as the parameters of antiprothrombin, thrombin and prothrombin time were investigated in serum of venous blood. In EE, the below parameters were found to be essentially increased: thrombin time--35.69 +/- 5.77 sec (controls--20.2 +/- 0.8 sec, p < 0.05), prothrombin time--30.43 +/- 7.21 sec (controls--21.6 +/- 0.8, p < 0.05), concentration of alpha2 macroglobulin--489.3 +/- 39.71 mg/dl (controls--240.2 +/- 12.2 mg/dl, p < 0.001), and AT-III activity--127.37 +/- 7.13% (controls--98.2 +/- 2.56, p < 0.001); at the same time, the parameters of plasmin were decreased- 108.00 +/- 5.28% (controls--169 +/- 18%, p < 0.01). PMID- 15881126 TI - [The neuromediatory system in liquidators of the radiation accident at the Chernobyl nuclear power station]. AB - The condition of mediatory systems, those of serotonin-, dopamin- and cholinergic, were studied in liquidators of the accident at the Chernobyl power station 15 year after the accident during the reaction of rosette-formation. The results showed an activated status of the studied serotonin- and cholinergic systems. PMID- 15881127 TI - [Involvement of non-clostridial anaerobic microorganisms in pathogenesis of chronic osteomyelitis]. AB - The microbic pattern of osteomyelitic foci were analyzed in patients with chronic osteomyelitis on the AB Expression Analyzer by the 'BioMerieux" anaerobic cultivation and identification system (France). The osteomyelitis-focus microcenosis was shown to be of the mixed anaerobic-aerobic nature. Anaerobic microorganisms were isolated in 65.4% of examined patients and made 34% of the isolated bacterial cultures, which is indicative of an important role of anaerobes in the development and persistence of the purulent-inflammatory process in chronic osteomyelitis. Thus, it is feasible to prescribe the antibacterial drugs, which are active in respect to both aerobic and anaerobic bacteria. PMID- 15881128 TI - [Perinatal aspects of serological diagnosis in urogenital chlamydiosis]. AB - The pregnancy course, labor, delivery and early neonatal period were studied in pregnant women with Chlamydia trachomatis infection as well as in their children. The levels of specific antibodies to Chlamydia trachomatis pgp 3 plasmid protein in maternal sera as well as perinatal complications were comparatively analyzed. According to the results, examination of anti-pgp3 antibodies is a highly specific and useful tool for the detection of Chlamydia trachomatis infection at its active stage in pregnant women. PMID- 15881129 TI - [Methods of extraction of Mycobacterium tuberculosis DNA from clinical samples for PCR: comparison and evaluation]. AB - Several commonly used methods of extraction of Mycobacterium tuberculosis DNA from clinical samples for further use in PCR were compared. We used the commercial sets "DNA-express" (Litekh, Russia) and "DNA-sorb-B" (Amplisens, Russia) as well as phenol extraction, fast alkaline lysis and DNA sorption on silicone gel for DNA extraction. All methods were compared by the quantity of isolated DNA and stability of DNA preparation as well as by efficiency and cost of extraction. "DNA-express" proved to be most effective in the extraction of Mycobacterium tuberculosis DNA with its cost being the smallest. PMID- 15881130 TI - [Human mycoplasma and mycoplasmic infections (Lecture, Part II)]. PMID- 15881131 TI - [International standard ISO/FDIS 15189]. PMID- 15881132 TI - [New European directives for diagnostics in vitro]. PMID- 15881133 TI - [Informational provision in the forensic medical service management: notion and structure of the information resources]. PMID- 15881134 TI - [Ascertainment of the cause and tempo of death in forensic medical practice]. AB - In order to develop a method of ascertaining a cause of death and of evaluating its tempo the authors undertook a histological study and a study of a dehydration degree in 76 cases of death due to the below reasons: mechanical trauma, mechanical asphyxia, burn shock, and carbon monoxide intoxication. Acute emphysema of lungs, lack of edemas in lungs and brain, desquamation of pulmonary macrophages, hyperemia of renal capillaries, primary urine in some glomerules, prevalence of ischemic changes with karyopyknosis in medulla and hypohydration of brain were typical of fast death. Prevalence of dystelectasis in lungs, emptying of pulmonary capillaries, evident edema of lungs and brain, desquamation of pulmonary macrophages, lack of primary urine in glomerular capsules, acute circulatory disorders of renal hemodynamics with sweating of fibrin and erythrocyte glomerules into cavities, necrotic nephrosis, pigment cylinders in renal tubules, thrombi in vessels, prevalence of chromatolysis, karyolysis and cytolysis in medulla, pronounced glial reaction as well as pronounced edema of brain and its hyperhydration were typical of slow death. PMID- 15881135 TI - [Histomorphometric characteristic of human brain in acute alcoholic intoxication]. AB - Different brain sections were studied in 20 subjects, who died of ethanol intoxication and in 14 subjects who died of injuries of the heart and main vessels, in order to detect histological changes in the brain and for the purpose of defining spatial and quantitative ratios between cerebral tissue structures in alcoholic intoxication. Different histological, stereometric and morphometric tools were made use of. It was demonstrated that, in alcoholic intoxication, there occur severe disorders of the circulation with affection of vessels in the brain; there are also dystrophic and necrotic changes in neurocytes, glial cells and white substance. The square of neurons shrinks due to death of some of them in the cortex of hemispheres, thalamus and cerebellum. As for the medulla, they are more resistant, there, to ethanol. The diameter of capillaries in the studied brain sections diminishes due to a reduced tonus of cerebral arteries; the quantity of such vessels increases within a standard area, which is conditioned by the compensatory opening of reserve capillaries. All this can be important in dealing with issues of thanatogenesis and of forensic medical diagnosis in death of alcoholic intoxication. PMID- 15881136 TI - [Diagnostic criteria in acute ethanol intoxication as cause of death]. AB - There are described, in the paper, morphological, chemical and biochemical signs in acute alcoholic intoxication. A complex approach is suggested to diagnosing an acute alcoholic intoxication as death reason. PMID- 15881137 TI - [Thanatogenesis in poisoning with psychopharmaceuticals]. AB - Discussed in the paper are thanatologically significant morphological signs in intoxication with some of psychic drugs. It was demonstrated that, in intoxication with azaleptin, phenazepam and aminazin, death comes mostly from affection of the brain including its edema and irreversible severe changes of neurons. In intoxication with tizercin, benzodiazepines and barbiturates, cardiac thanatogenesis as fibrillation of heart chambers combined with foci of myocytolysis. Sodium oxybutyrate brings about vascular collapse with subsequent development of fibrillation of heart chambers and of pulmonary edema. Intoxication with amitriptyline causes naturally necrotic nephrosis entailing uremic pneumonia and edema of the brain and heart. PMID- 15881138 TI - [Medico-criminal examination of damages to clothes in flight accidents]. AB - Examination of cloths is a permanent sphere in the practice of forensic-medical expert and an indispensable expertise stage providing for decision-making on the key issues set before the expert. Although, the examination of cloth in forensic medical expertise has been productive, research in this field slowed down during the recent decades; definition of new sets of evident informative signs has been also suspended. The methodological potentialities of examination of cloths items were demonstrated by examples of aviation incidents because such incidents reflect the principle variations of impacts, i.e. blow, explosion, fire, compression and chemical factors etc., which are subject to examination in the forensic-medical practice. Identification of mechanism causing damage and other signs on cloths helps a lot the forensic-medical expert in solving quite a number of issues related with circumstances of an incident and with a nature of an impact encountered by a person. PMID- 15881139 TI - [Ax and Am antigens: possible diagnostic reagents to them]. AB - Antigens of the discussed groups are difficult to diagnose; they have some features of group O and can cause expertise errors. Elucidated in the paper are issues related with determination of such antigens, in particular, by means of phytohemagglutinins of anti-(A+B). PMID- 15881140 TI - [Research of potentially linked variation of polymorphism of chromosome DNA in aspect of forensic expertise using molecular-genetic individualizing systems CD4, vWA and vWFII]. AB - Investigated within the case study are parameters of disbalance of lineage (HC) for 4 micro-satellite locuses of human genome: LPL, CD4, vWA and vWFII. The above locuses are widely used, both in Russia and abroad, in molecular-genetic applications for personality identification. Meanwhile, according to cytogenetics criteria, CD4, vWA and vWFII, are located close to each other in the telomeric region 12pter-12p12 in the short chromosome 12 arm, therefore their potential genetic interdependence is still a topical issue. We found a reliable HC between locuses vWA and vWFII. Locus CD4 did not display HC with locuses vWA and vWFII or with locus LPL. The latter, which is located in chromosome 8 and which must have been negative control for HC, was shown to have no HC with any of the studied markers. Such results correlate well with data on the relative physical localization of CD4, vWA, vWFII and LPL. Multiplication of frequency of alleles (genotypes) is not acceptable in typing locuses vWA and vWFII within one multi locus panel due to the genetic linkage of these markers demonstrated within the present case study, which is an important practical conclusion. PMID- 15881141 TI - [Detection of cannabinoids in urine]. AB - Described in the paper are results of comparative examination of urine by the methods of immunochromatographic express analysis (ICA), fluorescence polarization immune analysis (FIA) and of thin-layer chromatography (TLC) made for the purpose of detecting consumers of hemp products. A high specificity and a good sensitivity of the methods were demonstrated, which is a basis for using them as preliminary tests in detecting cannaboids and their metabolites in urine. The methods were evaluated quantitively and qualitatively versus the etalon method of chromato-mass-spectrometry. A possibility was pointed out to apply the methods of ICA and TLC as system analysis in the determination of 11-nordelta-9 tetrahydrocannabinol-9-carbonic acid in urine of marijuana users. PMID- 15881142 TI - [Detection of reladorm and donormil in forensic-medical examination of cadavers]. AB - A method was suggested for identifying reladorm and donormil in pharmaceutical drugs and biological objects. The above substances are isolated by 96% ethanol or by mixture of chloroform and isopropanol. 7 color reactions, 3 microcrystalloscopic reactions and chromatography in thin sorbent layer are suggested for identification. PMID- 15881143 TI - [Detection of maprotiline and proserine in forensic medical examination of cadaver]. PMID- 15881144 TI - [Hypoxic conditions of newborns and infants and their forensic medical value]. PMID- 15881145 TI - [Organizational aspect of medical care rendered to children with abnormal refraction under the conditions of school education]. AB - The main purpose of the case study was to investigate the efficiency of different method treatment of children with abnormal refraction. One hundred and fifty children were examined and shared between 3 experimental and one control groups. Routine ophthalmic examination methods were made use of; they are: visometry, autorefractometry, ophthalmobiometry, determination of the scope of relative accommodation, biomicroscopy, ophthalmoscopy; additional examinations, like rheoencephalography and consultation by neurologist, were also used. It was for the first time that the efficiency of infrared pneumomassage and of color-pulse therapy in the treatment of abnormal refraction was dynamically studied under the conditions of school education (vision correction rooms). The amplitude of relative accommodation increased in 71.6% of cases and the acuity of vision--in 78.8% of cases after the main treatment course; the refraction stabilized in 25 30% of cases. The results declined in 25-30% of cases after 3-4 months, therefore, treatment courses need to be repeated 3 times per year. Besides, positive results were obtained in the treatment of abnormal refraction by punctate massage of biologically active points, by general exercise and exercise for eyes as well as by massage of the collar zone and by medicamental therapy. Indications and contraindications were defined for the complex treatment of accommodation cramp and of abnormal refraction. PMID- 15881146 TI - [Progressing myopia in children: does it need treatment or not?]. AB - The purpose of the case study was to evaluate the remote consequences of a complex of laser and repeated surgical sclerorestorative procedures made in progressing myopia and its complications. Three hundred and forty-six children, aged 8-10, with rapidly progressing uncomplicated myopia of 4.25 to 9.5 D were shared between 2 groups. Two hundred and forty patients of the experimental group were made sclerorestorative procedures and transscleral low-energy laser stimulation of the ciliary muscle by means of infrared laser MACDEL-09. No such treatment was applied to patients of the control group. When indicated, preventive laser coagulation of the retina was made in both groups. The dynamic 10-year follow-up over the status of refraction and eye bottom showed that the complex scheme of repeated sclerorestorative procedures and low-energy laser treatment combined with preventive peripheral laser coagulation of the retina cut the rate of progressing myopia and prevented peripheral vitreoretinal dystrophy and retinal detachment in children and teenagers with progressing myopia. PMID- 15881147 TI - [Clinical presentation, diagnosis and treatment of strabismus combined with ptosis]. AB - On the basis of examination of 172 patients with strabismus concomitant with ptosis, different clinical forms of the former were defined and their clinical variations were classified. Symptomatology was described for each form. An independent type of false positive ptosis was pointed out. The notion of "positive and negative fixation tests" was introduced. A proper interpretation of the origin of true and false ptosis was presented. Finally, a sequence and tactics of surgical and functional treatment of different clinical presentations of squint concomitant with ptosis are lain down. PMID- 15881148 TI - [Diagnosis and treatment of children with acute inflammatory diseases of the orbit]. AB - The role of computer tomography within the differential diagnosis of acute inflammatory lesions of orbit tissues as well as an appropriate tactics of treatment were evaluated. Computer tomography, "Somafom AR-T", 512 x 512 pix matrix, Siemens (Germany), was made in the axial and coronary projections to delineate the inflammatory zone. The findings of computer tomography denoted explicitly the spatial features of pathology within the orbit with anatomic structures. In periostitis and reactive edema in retrobulbar fat of the orbit, the treatment of patients was conservative made jointly with otolaryngologist. Orbitotomy with drainage or with lancing of affected sinuses was made in abscess and phlegmon of the orbit. There was a complete recovery in a majority of patients with acute diseases of the orbit; they preserved their functions of the organ of vision and of its appendages. Therefore, computer tomography is a valuable and highly effective examination tool within the differential diagnosis of acute inflammatory diseases of the orbit. The diagnosis and treatment of such diseases call for a board of doctors including ophthalmologist, otolaryngologist and oral surgeon. PMID- 15881149 TI - [Ophthalmologic symptomatology in children with periventricular leucomalation]. AB - Ophthalmologic symptoms were analyzed in 76 children with periventricular leucomalation (PL). Clinical or functional ophthalmic disorders were detected in 100% of patients. Changes in the optic nerve disc were diagnosed in 93.4% of patients. The syndrome of dilated excavation (SDE), i.e. dilation and cupping of the optic nerve disc combined in all cases with affection of postgenicular visual paths (of optic radiation and/or striatal cortex) verified by neuroradiology, was most frequently (80.3%) encountered in children with PL. Presumably, the progression of SDE is associated with transsynaptic retrograde degeneration conditioned by the hypoxic-ischemic affection of the CNS in the projection of postgenicular visual paths at pre- and perinatal stages. SDE is an important criterion in the diagnosis of lesions of postgenicular visual paths in babies. PMID- 15881150 TI - [Polypeptide bioregulators in the treatment of different-type abiotrophy of the retina]. AB - The efficiency of retinalamin was studied in 33 patients, aged 4 to 7, with 5 types of retinal abiotrophy. The experimental group comprised children with retinal abiotrophy of Franceschetti type (36%) and mixed type (39%). The pathological process was stabilized during 18 months in all cases of developed disease. The visual acuity improved in 82% of patients after treatment by retinalamin; visual fields enlarged and central scotomas reduced in 54% of cases. Parameters of elctroretinography improved in 37% of cases. PMID- 15881151 TI - [Reactivation of persistent herpes virus infection as a factor of endogenous uveitis in children]. AB - Two hundred and fifty-five children with endogenous uveitis, aged 2 months to 15 years, were examined. 73-93% of children were chronically infected by different viruses of the human herpes group; mycoplasmal past-infection was detected in only 13% of them and Chlamydia past-infection _ in 3.7% of them. Herpes simplex virus of type 1 reactivated reliably more often versus other types of Herpesviridae. Reactivation of cytomegalovirus infection prevailed in mothers who gave birth to children with intrauterine uveitis. A prolonged active replication of herpes virus was primarily observed in children with a Suppressed cell antiviral immunity component. Uveitis in such children was notable for a severe clinical course and trend towards often relapses. PMID- 15881152 TI - [Pavlyuk--antioxidant "Gistochrom" in the complex treatment of intraocular hemorrhages in children]. AB - We studied the efficiency of Russian antioxidant Gistochrom in the treatment of intraocular hemorrhages of different genesis in children. Gistochrom was used in 407 children (449 eyes), aged 1 month to 14 years, with traumatic hyphema, hemophthalmos, hemorrhages in the eye bottom, hemorrhagic retinovasculitis, recurrent hephema, diabetic retinopathy, hemorrhages after reconstruction of the anterior chamber with cataract extraction, neuritis, retinopathy of prematures as well as with uveitis with a pronounced exudative component, with signs of retinal edema and of reactive pappilitis. Gistochrom has an intense action on blood resorption in structures of the eye ball as observed in traumas and lesions of internal eye tunics with hemorrhages. The treatment efficiency is higher when the drug is used at earlier disease stages and when it is administered into the retrobulbar space through the irrigation system. Gistrochrom is recommended for use in ophthalmic practice for the treatment of intraocular hemorrhages of different genesis as well as in retinopathy with a pronounced exudative component and in different-etiology uveitis. PMID- 15881153 TI - [Irrigation therapy as a method of intensive care of ophthalmopathology in the posterior eye segment in children]. AB - Presented in the paper are data of a comparative analysis of efficiency of different methods of administration of drugs in neuritis and partial atrophy of the optic nerve. New techniques of application and fixation of irrigation systems in the retrobulbar and Tenon's space are described. Experimental and clinical data proving advantages of the new method of administration of drugs by an automatic pulse doser in the treatment of inflammatory diseases of the optic nerve are represented. The use of such intensive intermittent technique of administration of drugs in Tenon's space performed at the preliminary stage before electrostimulation of the optic nerve made the procedure by far more effective and ensured better treatment results. PMID- 15881154 TI - [Laser coagulation in the treatment of different-type retinopathy of prematures]. AB - Laser coagulation of the retina was made in 126 children (238 eyes) with retinopathy of prematures (RP) by an 810 nm diode laser ophthalmocoagulator. The methods of transscleral, transpapillary and combined coagulation were applied. As for children who needed coagulation, 39.9% of them had classical RP and 60.1%- fulminant RP (plus-disease). Stabilization in CRP was registered in 97.9% (the threshold process stage was an indication for surgery), and in FRP (plus disease) it was registered in 56.6%. The best effect from surgery in FRP was observed, when patient were operated on in FRP, stage 1 (process localization in zone 1) and stage 2 (process localization in zone 2), whereas, when the procedure was made at stage 3, the process was stabilized only in 37%. The authors elaborated a combined method of laser coagulation (including transscleral and transpupillary techniques), which brought up the rate of stabilization in FRP (plus-disease) to 75.8%. The results did not depend on a coagulation technique in CRP. PMID- 15881155 TI - [An eight-year experience of cryotherapy of acrive retinal retinopathy in prematures: management of patients and results]. AB - The paper represents an eight-year experience of active retinopathy of prematures (RP) by the method of transscleral cryopexy of the retinal zone in 91 patients (181 eyes) The time period and periodicity of ophthalmologic follow-up of prematures with RP risk are defined; indication and contraindications for cryotherapy are formulated; complications observed at the early postoperative period are pointed out. Cryotherapy was shown to be an effective method of treatment in RP: favorable treatment results were registered in 54.5% of cases. There are also results of a comparative analysis of the clinical refraction condition observed in RP patients in 4.8-5 years after cryotherapy. Myopia was by far more often diagnosed in the group of RP patients (38.2%). This parameter made 12% (p < 0.001) in the group of equal prematures. PMID- 15881156 TI - [Problems and outlooks for the preventive treatment of retinopathy in prematures]. AB - The immediate and remote (up to 7 years) results of preventive cryo- and/or lasercoagulation of avascular retinal zones made in 390 children with active progressing retinopathy of prematures (RP) were analyzed. The efficiency of cryocoagulation in processes localized in zones 2-3 of the eye bottom made 92%, and that of the combined technique (laser- and cryocoagulation) with lesions in zones 1-2 and with plus-diseases was up to 80%. The preventive treatment is indicated for the 3d threshold (ordinary clinical course) and before-threshold (with plus-disease and localization zone 1) stages. Such treatment is not justified when undertaken early and in developed retinal detachment. Further research of the disease pathogenesis is needed to enhance the efficiency of the RP treatment, i.e. research of growth factors of endothelial vessels, autoimmune reaction to S-antigen of the retina and of dosed oxygentherapy etc. PMID- 15881157 TI - [Surgical treatment of traumatic lesions of the orbit in children]. AB - The paper presents an analysis of traumas of the orbit combined with craniocerebral trauma in children made according to data obtained at the eye microsurgery department, Morozov's Pediatric City Clinical Hospital. Fractures of the lower orbit wall account for around 70% of all orbit fractures; clinical signs of fractures of the lower orbit wall are defined; computer tomography is shown to be the most informative diagnostic tool. The paper contains indications for plasty of the lower orbit wall. Materials used in orbitoplasty are comparatively analyzed. The use of demineralized osseous implants is preferential. PMID- 15881158 TI - [The specificity of surgery of congenital cataract with implantation of folding "Acrysof" IOL in children]. AB - Described in the paper are the results of surgical treatment of 25 children (37 eyes), aged 3 months to 12 years, with congenital and secondary cataracts. The treatment schemes were based on new technologies including tunnel incisions, continuous round anterior capsulorhexis and aspiration-irrigation as well as implantation of different-model posterior-chamber flexible "Acrysof' IOLs into the capsular sac or ciliary sulcus. A high treatment efficiency as well as a need in a differentiated approach to choosing an IOL fixation, diameter and technique of anterior capsulorhexis and a model of the implanted "Acrysof' IOL made with respect to individual age and clinical peculiarities of eyes of children with congenital cataracts as well as with respect to a potential general somatic pathology (galactosemia) are demonstrated. PMID- 15881159 TI - [The state-of-the-art of surgical treatment of congenital cataract in prematures]. AB - Described in the paper are results of a clinical treatment of 127 children (174 eyes), aged 2 to 15 years, with different-type congenital cataract; the treatment was based on modem technologies including tunnel limbic incision, continuous anterior round capsulorhexis, aspiration-irrigation of the contents of the capsular sac, implantation of soft Acrysof SA30A1 IOLs by forceps or "Monarch" injector and suturing of incision. The anterior continuous capsulotomy was found to be the most complicated surgery stage in removal of congenital cataract in children it could be made without errors in 75.3% of cases. Intracapsular implantation of IOL in different-type cataract in children was possible in 97.7% of cases. The single-unit Acrysof IOL stays stable inside the capsular sac for as long as 24 months. A combination of primary capsulorhexis with "dry" anterior vitrectomy is most effective in the prevention of secondary cataract. PMID- 15881160 TI - [Treatment of retinoblastoma: modern concept]. PMID- 15881161 TI - [Unilateral pigment retinitis concomitant with congenital heart disease]. PMID- 15881162 TI - [A case of secondary orbital echinococcosis]. PMID- 15881163 TI - [Endoscopic diagnosis of colitic cancer and dysplasia]. PMID- 15881164 TI - [The concept, criteria for diagnosis, and disease classification of ulcerative colitis]. AB - Ulcerative colitis (UC) is an idiopathic, non-specific inflammatory disorder involving primarily the mucosa and submucosa of the colon, especially the rectum. It usually produces a bloody diarrhea and various degrees of systemic involvement. These definitions of idiopathic proctocolitis were established by Council for International Organization of Medical Science (CIOMS) in 1973. In Japan, the first criteria for diagnosis of UC were established by the Research Committee of UC (the Japanese Ministry of Health and Welfare) in 1974, based on the definitions by CIOMS. Since then, some minor revisions have been made. The newest criteria for diagnosis of UC were published in 1998. The diagnosis of UC is made on the basis of a combination of clinical, endoscopic, radiologic, and histologic findings. PMID- 15881165 TI - [Epidemiology of ulcerative colitis--comparison between Japan and western countries]. AB - Epidemiological data of ulcerative colitis both in Japan and other countries were presented. The prevalence has been increasing in Japan in the past few decades, although it is still lower than that reported from western countries. The incidence in Japan as well as other countries in Asia is increasing. On the other hand, the incidence in western countries has been reported to be decreasing in some reports. There seems to be no significant difference in age of onset between Japan and western countries. Cause of death in patients with ulcerative colitis has changed in Japan after 1980. Death related with colorectal cancer has been increasing in the past 25 years in Japan. PMID- 15881166 TI - [Etiopathogenesis and aggravating factors in ulcerative colitis]. AB - Ulcerative colitis (UC) is a chronic inflammatory disease limited to the colon. Although the pathogenesis of inflammatory bowel disease (IBD) remains unclear, several studies have suggested that the onset and development of IBD require the interaction between genetic susceptibility, stimulation by luminal bacterial antigens and adjuvants, and episodic environmental triggers which break the mucosal barrier. There are many reports that experimental enterocolitis in animals does not occur in a sterile (germ free) environment and is prevented by antibiotics therapy. Moreover, patients with UC exhibit pathological immune responses to many commensal enteric bacterial species. Recent data showed that certain probiotic species decrease relapse of UC. These findings suggest that the most possible cause of UC is associated with chronic intestinal inflammation which is induced and perpetuated by non-pathogenic bacteria in genetically susceptible hosts. PMID- 15881167 TI - [Pathomorphology of ulcerative colitis]. AB - Microscopic and macroscopic appearances of ulcerative colitis (UC) by its phase of inflammation were summarized. The most characteristic microscopic findings of active phase UC is diffuse lymphoplasmacytic infiltration, essentially associated with basal plasmacytosis. Although inflammation of UC is basically limited to the mucosa, active inflammation extends into the submucosa in some instance, and acute ischemic change is overlapped to cause toxic megacolon. In remission phase, inflammation is reduced and goblet cell mucus is fully recovered but evidences of the past inflammation such as irregular shape and disarrangement of crypts, Paneth cell metaplasia, thickening of the muscularis mucosae and discrepancy between the crypt base and the muscularis mucosae are usually demonstrated. Macroscopic appearances of UC reflect its microscopic findings such as degree of inflammation, whether inflammation is (was) limited to the mucosa or extend(ed) into the submucosa. Active phase is classified into erythematous, spongy, granular, pseudopolyp, ulcerative, and fulminant (toxic megacolon) type. In the former two types, inflammation is limited in the mucosa, and the latter two types are associated with ischemic change. In remission phase, erythematous and spongy types recover to the almost normal looking mucosa or fine granular mucosa with preservation of mucosal folds, granular type recovers to granular, fine granular or flat atrophic mucosa without preservation of mucosal folds, and pseudopolyp type recovers to mucosa with inflammatory polyposis. PMID- 15881168 TI - [Genetic and immunological basis for ulcerative colitis]. AB - Ulcerative colitis is a chronic inflammatory disease of the rectum and colon. Results from many studies in people and animals of intestinal inflammation suggest that ulcerative colitis results from environmental factors triggering a loss of tolerance for normal intestinal flora in genetically susceptible individuals. Although progress has been made in the overall management of the disease, there are few clinical data on biological agents in contrast to Crohn' s disease. Here, we discuss the genetic and immunological basis of ulcerative colitis including the recent findings. PMID- 15881169 TI - [Probiotics for inflammatory bowel disease]. AB - Probiotics are defined as live microorganisms that may beneficially affect the host by improving the balance of the intestinal micro flora. In spontaneous animal models of inflammatory bowel disease (IBD), the influence of the enteric flora on gut inflammatory activity has been demonstrated, as germ free animals do not develop disease. Modification of the activity of the intestinal flora using probiotics has been investigated as a way of controlling IBD. However, there is still considerable work to be done before probiotics can be considered as part of the standard treatment of IBD. Probiotic cocktails should be carefully studied and the individual bacterial constituents evaluated in clinical settings. Several studies are currently underway to assess these issues. Further study of placebo controlled trial testing therapeutic role of probiotic bacteria in human trials is warranted. PMID- 15881170 TI - [Radiological diagnosis of ulcerative colitis]. AB - In cases with ulcerative colitis, barium enema is valuable in objectively determining the stage (active, inactive) and defining the extent and aggressiveness of the disease that often varies during the course. The preparation, findings, and differential diagnosis of the technique are reviewed. PMID- 15881171 TI - [Diagnostic principles of colonoscopy in ulcerative colitis]. AB - Colonoscopy plays a most important role in the diagnosis and management of ulcerative colitis (UC). It is generally performed in UC for defining the degree of disease activity, grading endoscopic disease severity, and surveillance to detect early cancer and/or dysplasia. Colonoscopic findings of UC are characterized by diffuse and continuous inflammation of mucosa. Loss of vascular pattern, erythema, petechiae and mucosal friability are features of relatively mild disease. As the severity of inflammation increases, spontaneous bleeding, erosions and/or ulcerations are observed. Although UC can be easily diagnosed by these typical findings, colonoscopic findings are modified in a variable and complicated manner at the different clinical stages. Thus it is important for endoscopists to understand the findings in each clinical stage of UC. PMID- 15881172 TI - [Histopathology of ulcerative colitis in biopsy]. AB - For histologic diagnosis on biopsy specimens, we gave an outline of macroscopic and microscopic features of ulcerative colitis (UC), including differential diagnosis. The most important histologic features on biopsy are dense mixed inflammatory infiltrate in the lamina propria mucosae, mucosal architectural abnormality, and vascular change. Evidence of colorectal adenocarcinoma, dysplasia, and cytomegalovirus infection should also be taken into account. Although diagnostic criteria for UC are well understood, the biopsy diagnosis of UC is comparatively difficult. Thus the diagnosis of UC should be made mainly by clinical pictures, course of the illness, and radiological and endoscopic pictures. PMID- 15881173 TI - [Ultrasonography for diagnosis of ulcerative colitis]. AB - Ultrasonography (US) is a useful tool in the assessment of activity in ulcerative colitis (UC). Different studies have reported that US can correctly define the extension of UC, and the degree of bowel wall thickness is correlated to the clinical activity. The examination with a linear transducer in tissue harmonic imaging is indispensable to the detail evaluation of bowel wall stratification. In the Doppler mode, widespread mural vessels are found in active UC. Without contrast agent, Advanced Dynamic Flow can depict tiny vessels in the submucosa with an Aplio80 scanner (Toshiba, Japan), and improve the measurement of arterial blood flow velocity. Therefore, US should be considered as a reliable method for the evaluation of disease activity in UC. PMID- 15881174 TI - [Carcinogenesis in ulcerative colitis-associated colon cancer and surveillance colonoscopy]. AB - It has been widely accepted that long-standing cases with ulcerative colitis, especially pancolitis have higher risk of complicating colitic cancer. Colitic cancer often appears as multi-focal invasive features with un-differentiated histology. Therefore, endoscopic diagnosis of precancerous or cancer associated dysplastic lesions. Surveillance colonoscopy has preventing effects for developing colitic cancer. In western world, annual surveillance colonoscopy with multiple stepwise has been adopted. In Japan, surveillance colonoscopy with targeted biopsies assisted by chromoendoscopy and/or magnifying endoscopy has proven to be efficacious in finding such lesions. PMID- 15881175 TI - [Clinical guidelines for the treatment of ulcerative colitis]. AB - Guideline for the treatment of ulcerative colitis was originally developed as a part of group studies for the inflammatory bowel diseases in 1975, supported by Japanese Ministry of Health and Welfare. It was revised almost annually by adopting newer knowledge. The outline of the latest version was presented in the text. Although recommendations within the guideline seem to be adequate, present guideline was developed based on the opinions of the experts. Future issues include standardized method in developing practice guideline with emphasis on clinical evidence, in order to maintain scientific validity. PMID- 15881176 TI - [Drug therapy for ulcerative colitis: salazosulfapyridine and 5-ASA]. AB - Aminosalicylates have a wide range of anti-inflammatory and immunomodulatory effects. Oral salazosulfapyridine (SASP) and 5-aminosalicylic acid (5-ASA) are the 'first-line' therapy for induction of remission in mild to moderate active ulcerative colitis (UC). SASP, which is consisted of 5-ASA and sulfapyridine, has greater incidence of side effects. 5-ASA is a therapeutically active compound, while sulfapyridine is related to adverse effects. For this reason, 5-ASA formulas exclusive of sulfapyridine were developed and they enabled higher dose of 5-ASA administration without adverse effects. Topical treatment by 5-ASA enema or SASP suppository should be considered for the treatment of proctitis or distal type of UC. Oral aminosalicylate therapy is also effective for the maintenance of remission in UC. Therefore, aminosalicylates are key drugs for the treatment of UC. PMID- 15881177 TI - [Glucocorticoid treatment in UC]. AB - Topical or systemic administration of glucocorticoid (GC) plays major role in UC therapeutic strategy. For severely-active cases, more than 60 mg/day intravenous regimen has been recommended. Oral prednisolone at 30-40 mg/day dose for moderate severity of UC, or GC enema for localized UC are also used in general practice. However, it is proved by controlled trial that low-dose GC has no effect on preventing the relapse of UC. To improve the QOL in UC patients, all administrated GC must be tapered off after achieving the remission. GC receptor expression is essential for pharmacophysiological effects of GC on target cells. It is reported that beta isoform of the GC receptor contributes the unresponsiveness to GC. Prediction of GC-sensitivity by measuring the expression of GC receptor isoforms in PBMC of UC patients is also discussed. PMID- 15881178 TI - [Immunosuppressants for therapy of patients with ulcerative colitis]. AB - Ulcerative colitis (UC) has been known as inflammatory bowel disease. Progress in UC management strategies has led to optimized approaches for achieving the two primary clinical goals of therapy: induction and maintenance of remission. We here review about immunosuppressants in management of UC; Cyclosporine A (CsA) has been used for the induction therapy in steroid resistant refractory UC. Although it has been reported that CsA has high response rate in severe UC, long term efficacy (maintenance of remission) has not been proven. To improve maintenance therapy, immunosuppressant has been re-considered in management of UC. In recent years, it has been reported that efficacy of 6 mercaptopurine/azathioprine in maintenance of remission of UC is superior to 5 aminosalicylate (5-ASA). Pharmacological studies have indicated thiopurine methyltransferase (TPMT) activity is essential for maintenance of blood concentration of 6-thioguanine nucleotide (6-TG). PMID- 15881179 TI - [Cytoapheresis for IBD]. AB - Ulcerative colitis (UC) is associated with chronic relapsing inflammation of the colon. Cytoapheresis is a therapeutic strategy of extracorporeal immunomodulation that has been used in several immunological disorders. In Japan, several open trials of cytoapheresis for UC patients accumulated encouraging results in bringing steroid-resistant patients into remission. In this paper, three types of cytoapheresis [leukocytapheresis (LCAP), granulocytapheresis (GCAP), and centrifugal leukocyte apheresis] are reviewed. PMID- 15881180 TI - [Enema therapy for ulcerative colitis patients]. AB - The therapeutic options for the treatment of mildly to moderately active distal colitis or distal flare up of total colitis have increased over the last decade. Many reports have documented the effectiveness of various compounds, such as 5 aminosalicylic acid, steroids, cytoprotective agents, lidocaine gel, short chain fatty acid, and cyclosporine when they are applied topically to the inflamed distal colon in patients with ulcerative colitis. These treatments have considerably attracted our attention because of their possible anti-inflammatory effects on ulcerative colitis with lower toxicity. Larger, controlled studies will be necessary to establish the roles of enema therapy in patients with distal colitis. PMID- 15881181 TI - [Probiotics and prebiotics for the treatment of inflammatory bowel disease]. AB - Although the causes of inflammatory bowel disease including ulcerative colitis and Crohn's disease remain incompletely understood, increasing evidence implicates intestinal microflora in the pathogenesis of this disorder. Therefore, modulation of microflora with probiotics or prebiotics may offer a plausible therapeutic approach. While recent data support a potential therapeutic efficacy, such treatments need to be further assessed by large scale studies. A better understanding of the intestinal microflora and the mechanisms of their action may help us to develop more effective treatment for inflammatory bowel disease. PMID- 15881182 TI - [Surgical treatment for ulcerative colitis--recent advances]. AB - Surgical indication for ulcerative colitis is fulminant colitis, intractability, cancer or dysplasia. New surgical indication should be established because new medical treatment such as leucocytoapheresis or intravenous cyclosporine treatment developed. Standard surgical procedure is ileal pouch anal anastomosis with rectal mucosal stripping and stapled ileal pouch anal anastomosis. Postoperative bowel function and QOL are satisfactory in both of them. Surgical treatment should be performed without any delay for the patients who do not respond medical treatment. PMID- 15881183 TI - [Intestinal complications in ulcerative colitis]. AB - Although cases of severe intestinal complications such as massive hemorrhaging, perforation, stricture and obstruction occur in some instances of ulcerative colitis, these are relatively infrequent in comparison to Crohn's disease. Many cases with such severe intestinal complications require surgery, sometimes acute surgical emergency, instead of medication as these can be less effective in terms of improvement of the disease. Cytomegalovirus infections, which are at times associated with intractable ulcerative colitis, has recently been considered an etiologic factor in the exacerbation and intractability of ulcerative colitis. Furthermore, colitic cancer, an important complication of ulcerative colitis, may occur at high incidence in patients experiencing long term courses of chronically active or frequently relapsing ulcerative colitis, and thus must be considered and managed appropriately. For the prevention of complications of ulcerative colitis, rapid induction of remission and long-term remission maintenance is essential, in addition to the emphasis in avoiding chronic administration of large doses of steroids. PMID- 15881184 TI - [Extraintestinal complications in patients with ulcerative colitis]. AB - Extraintestinal complications of patients with ulcerative colitis contribute to quality of life and mortality of the patients. Extraintestinal complications involve many organ systems. Skin, mucocutaneous, hepatobiliary, and joints complications are major extraintestinal complications. Some disorders correspond the activity of the colitis, other complications develop independently of the bowel inflammation. The purpose of this review is to update the clinicians concerning the etiology, clinical course and the therapy of the extraintestinal manifestations associated with ulcerative colitis. PMID- 15881185 TI - [Evaluation and improvement of QOL among IBD patients]. AB - This article provided a short review on the assessment of quality of life (QOL) among patients with ulcerative colitis. Quantitative approach with psychometric measures has empirically showed that psycho-social aspects of patient's QOL are often affected by her clinical, psychological, and social conditions. Although the control of disease activities plays a pivotal role for improving patient's QOL, more holistic approach such as psychoeducational intervention should also be considered to enhance patient's self-efficacy and sense of control. Besides, qualitative research suggests that helping patient's narrative reconstruction through enhanced communication with the physician may be the most powerful tool that the physician can provide for improving patient's QOL. PMID- 15881186 TI - [Diagnosis and medical treatment of pouchitis in ileal pouch-anal anastomosis in patients with ulcerative colitis]. AB - Pouchitis is a common and long-term complication of ileal pouch-anal anastomosis in patients with underlying ulcerative colitis. Combination with clinical symptoms, endoscopic and histopathological findings are required to make accurate diagnosis. Ciprofloxacin and metronidazole are effective in patients with acute pouchitis. Probiotics may be useful in maintaining regression and preventing from remission. Immunomodulating agents are also useful in chronic pouchitis. PMID- 15881187 TI - [Guidelines for treatment of ulcerative colitis in children]. AB - Guidelines for treatment of ulcerative colitis in children have been created by the working group of the Japanese Society for Pediatric Gastroenterology, Hepatology, and Nutrition and the Japanese Society for Pediatric Inflammatory Bowel Disease. The ideas of these guidelines, with regard to the fundamental differences in medical treatment between children and adults, included (1) for children, intensive medical treatment including appropriate systemic management is important during the acute phase of illness. (2) Treatment with steroids, which can cause growth disturbances, should not be continued for long period of time. (3) Pulsed steroid therapy, selective removal of blood cells, and intravenous infusion of cyclosporin are included in the therapeutic option for severe and fluminant cases. PMID- 15881188 TI - [Lipid synthetic transcription factor, SREBP]. AB - SREBP family have been recently established as bHLH type transcription factors governing lipid synthesis. While SREBP-2 regulates expression of genes involved in cholesterol biosynthesis and LDL receptor, SREBP-1c controls fatty acid synthesis. Cellular cholesterol is regulated by feedback system where SCAP/Insig system regulates cleavage of SREBP-2 for its activation depending upon cellular cholesterol demand. Meanwhile, SREBP-1c is nutritionally regulated, excess energy intake could activate hepatic SREBP-1c expression leading to formation of remnant lipoproteins and hepatic insulin resistance through suppression of IRS-2 expression. Hepatic SREBP-1c could be deeply involved in metabolic syndrome. Thus, SREBP-1c could be a therapeutic target to treat metabolic syndrome. Polyunsaturated fatty acids suppress SREBP-1c and suppress lipogenesis. PMID- 15881189 TI - [Clinical significance of home blood pressure monitoring and its application to daily clinical practice]. AB - Seven million home blood pressure (HBP) monitors are manufactured annually and a total of more than 30 million monitors have been sold in Japan. HBP monitoring is regarded as clinically useful according to various overseas guidelines. The criteria for the diagnosis of hypertension based on HBP monitoring are specified by the Japanese Society of Hypertension (JSH) because of its excellent prognostic value, and it was also assessed as clinically useful in the 2004 JSH hypertension treatment guidelines (JSH 2004) released last autumn. Thus, HBP monitoring is becoming increasingly important. HBP monitoring boosts a patient's awareness of the treatment of hypertension, leading to better compliance with drug intake and overall treatment. In addition, it is very useful for assessing the therapeutic effect and duration of action of drugs. HBP morning is also required for the diagnosis of morning hypertension as well as white coat hypertension and reverse white coat hypertension. The HBP monitors released in recent years can even measure blood pressure during sleep. The JSH 2004 regarding HBP monitoring was based on "The 2003 Guidelines for HBP Monitoring". As mentioned above, HBP monitoring is expected to contribute to improving the treatment of hypertension and medical economy. Both the blood pressure measured at a hospital and that measured at home are red blood pressures. It is impossible to determine a single specific blood pressure value for a patient, so it appears to be more rational to estimate the usual blood pressure from data obtained under various conditions. In this article, we review the clinical meaning of HBP monitoring based on the current situation and discuss problems with HBP monitoring and its application to daily clinical practice. PMID- 15881190 TI - Simultaneous measurement of multiple radiation-induced protein expression profiles using the Luminex(TM) system. AB - Space flight results in the exposure of astronauts to a mixed field of radiation composed of energetic particles of varying energies, and biological indicators of space radiation exposure provides a better understanding of the associated long term health risks. Current methods of biodosimetry have employed the use of cytogenetic analysis for biodosimetry, and more recently the advent of technological progression has led to advanced research in the use of genomic and proteomic expression profiling to simultaneously assess biomarkers of radiation exposure. We describe here the technical advantages of the Luminex(TM) 100 system relative to traditional methods and its potential as a tool to simultaneously profile multiple proteins induced by ionizing radiation. The development of such a bioassay would provide more relevant post-translational dynamics of stress response and will impart important implications in the advancement of space and other radiation contact monitoring. PMID- 15881191 TI - Profile of the JACC study. AB - BACKGROUND: A large-scale population-based cohort study would offer the best evidence of a relation between lifestyle and cancer. METHODS: The Japan Collaborative Cohort Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho) was established and carried out from 1988 to 1990 in 45 areas in Japan. Epidemiological information, such as demographic information, past medical history, exercise/sports activities engaged in, frequency of food intake, smoking and alcohol drinking status and so on, was collected by a self-administered questionnaire. Blood samples were collected from each participant at screening in 37 out of 45 areas to investigate risk factors of cancer relating to biochemical substances in blood. Collected sera were divided into 3-5 tubes (100-500 microL per tube) and stored at -80 degrees C. Additionally, about 5 years after the baseline survey, an interim survey regarding lifestyle changes was conducted in 31 areas. We followed the study subjects for mortality, move-outs, and cancer incidence, if possible (in 24 areas). RESULTS: There were 127,477 participants (54,032 men and 73,445 women) registered in the study. Of these, 110,792 subjects (46,465 men and 64,327 women), aged 40 to 79 years at baseline, were eligible for follow-up. Sera were stocked from 39,242 subjects, and interim survey was carried out on 46,650 subjects. CONCLUSIONS: The JACC Study provides useful evidence for cancer prevention in Japan. PMID- 15881192 TI - Reproducibility and validity of a self-administered food frequency questionnaire used in the JACC study. AB - BACKGROUND: A self-administered questionnaire on dietary habits used in the JACC Study contained a 40-item food frequency questionnaire (FFQ). Although more than 110 thousand subjects enrolled in JACC Study and responded to the FFQ, no validation study has been conducted to date. METHODS: Eighty-five volunteers among the cohort members completed 2 FFQs (FFQs 1&2) and 12-day weighed dietary records (WDR). The interval between the two FFQs was one year. During the one year, the subjects carried out a 3-consecutive-day WDR in each season. We tested the reproducibility by using two FFQs. Also, we tested the validity of the FFQ by using the 12-day WDR as a gold standard. RESULTS: The intake frequencies of the 2 FFQs often agreed, showing the Spearman correlation coefficients ranging from 0.42 (edible wild plants) to 0.86 (coffee). The Spearman correlation coefficients of the energy and nutrient intakes from FFQ2, and that of the 12-day WDR were 0.20(energy) to 0.46 (animal protein, potassium). After adjusting the energy intake, the correlation coefficients showed 0.21(fish fat) to 0.51(animal fat). When classifying the FFQ2 and WDR by quartiles and examining the degree of agreement between the two methods, we obtained its median 30%. CONCLUSIONS: The FFQ is suitable to deal with a large group of subjects. However, since the energy and the amount of nutrient intake from this FFQ can not show the overall dietary intake situation, the subjects' dietary intake should be assessed by categories. PMID- 15881193 TI - Frequency of food intake and estimated nutrient intake among men and women: the JACC Study. AB - BACKGROUND: The aim of this study was to determine the frequency of food intake and estimated nutrient intake in the JACC study cohort. METHODS: The subjects were 46,465 men and 64,327 women aged 40-79 years who responded to the self administered food frequency questionnaire. We calculated the dietary intake of major nutrients by multiplying the frequency of consumption of each food with each portion size, estimated from a validation study. RESULTS: Women reported to more likely consume vegetables, seaweed, fruits, sweets, oolong-tea, western style-breakfast, and less likely to consume rice and miso-soup than men. Women reported less preference of salty foods and fatty foods than men. Compared with men, women had higher mean intakes of carotene and vitamin C, and lower intake of total energy, carbohydrate and sodium. The frequency of consumption of beef, chicken, dairy products, fresh fish, fish products, rice, and miso-soup increased with age in men, and that of vegetables, seaweed, beans, tofu, fruits, sweets, and green-tea increased with age in both sexes. Men aged 40-49 years had the lowest mean intake levels of crude fiber, calcium, iron, retinol, carotene, and vitamins A, C, and E. Women aged 40-49 years had the lowest mean intake levels of crude fiber, iron, and vitamins C. Women aged 70-79 years had the lowest mean intake levels of calcium, retinol, and vitamins A. CONCLUSIONS: Women had a more westernized dietary pattern than men. Elderly men had a mixture of unhealthy and healthy dietary patterns while elderly women generally had a healthier dietary pattern compared with younger persons. PMID- 15881194 TI - Levels of physical activity among participants in the JACC study. AB - BACKGROUND: Physical activity is thought to play an important role in the maintenance and promotion of health, and practical questionnaires assessing levels of physical activity are currently a widely used method in epidemiological research. METHODS: As a part of the Japan Collaborative Cohort Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho), we investigated the status of physical activity of cohort participants aged 40-79. A total of 110,792 participants (46,465 men and 64,327 women) completed the baseline survey from 1988-1990. Questionnaires concerning physical activity contained the following questions: 1) How much time per week on average do you spend engaging in sports or physical exercise? (at least 5 hours, 3-4 hours, 1-2 hours, little); 2) How much time per day on average do you spend walking either indoors or outdoors? (longer than 1 hour, 30 min-1 hour, about 30 min, little). RESULTS: In total, 68.7% of men and 76.2% of women responded with "little" to the amount of time spent engaged in sports or physical exercise. The proportion of time spent on habitual physical exercise (sports and physical exercise of one hour or more per week) was lowest in the 50-59 years age group and increased with age among participants aged 50 79. In addition, 49.4% of men and 50.7% of women responded with "more than 1 hour per day" to the amount of time spent walking. CONCLUSION: These results show that the proportion of habitual physical exercise was relatively low in the JACC Study and increased with age among participants aged 50-79, whereas almost half walked for more than one hour a day. PMID- 15881195 TI - Change in food intake frequency at five years after baseline in the JACC study. AB - BACKGROUND: In a cohort study, information on an individual is taken at baseline, after which it usually remains fixed. There is some risk that this will lead to misclassification and cause weakened or biased results. To prevent such distortion, following up of exposure is important, although it is still scarce in practice. METHODS: In the Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study) sponsored by Monbusho (Ministry of Education, Science, Sports and Culture of Japan), 37,838 (14,531 males and 23,307 females) subjects out of a cohort of 127,477 inhabitants answered an interim questionnaire on food intake frequency consisting of 33 items about five years after registration. The long-term reproducibility was assessed using Spearman's correlation coefficients and agreement. From data at two time points, longitudinal change, age effect, and secular trend were examined. Subjective changes in these items at the time of the interim survey were also compared to longitudinal changes. RESULTS: Spearman's correlation coefficients varied from 0.27 (fruit juice in males) to 0.55 (beef in females and milk in males), and agreement from 29.9% (fruit juice in males) to 61.4% (liver in females). Correlation was relatively stronger in meat and dairy products and weaker in vegetables and fruits. In both males and females, most increased food item was edible wild plants followed by confectioneries (males) and yogurt (females). CONCLUSION: Over five years, food intake was considerably changed. These interim data could be used for a long-term follow-up study to prevent the results becoming weakened or biased. PMID- 15881196 TI - Smoking and drinking habits five years after baseline in the JACC study. AB - BACKGROUND: Observing longitudinal changes in smoking and drinking habits is important for evaluating the risk of incidence and death from cancer or other diseases in a cohort study. METHODS: Smoking and drinking habits at baseline and about five years later among 18,312 males and 28,338 females were obtained from the baseline and interim surveys in the Japan Collaborative Cohort Study for Evaluation of Cancer Risk (JACC Study) sponsored by Monbusho (Ministry of Education, Science, Sports and Culture of Japan). Changes in smoking and drinking habits between the two surveys were observed. Odds ratios for quitting these habits at interim survey were estimated. RESULTS: Percentages of current smokers at baseline and interim surveys were 51.0% and 45.5% in males, and 5.2% and 4.8% in females, respectively. Percentages of current drinkers at baseline and interim surveys were 78.0% and 73.2% in males, and 29.5% and 23.5% in females, respectively. The number of cigarettes per day among male current smokers and the usual amount of alcohol consumed on each occasion among current drinkers decreased between the two surveys. Odds ratios for smoking cessation increased with age at baseline and decreased with the number of cigarettes per day at baseline. Odds ratios for drinking cessation increased with age at baseline and decreased with the usual amount of alcohol consumed on each occasion at baseline. CONCLUSION: The decrease in smoking and drinking habits was observed during the five-year follow-up period. Higher age and lower levels of exposure were associated with quitting smoking or drinking. PMID- 15881197 TI - Stability of frozen serum levels of insulin-like growth factor-I, insulin-like growth factor-II, insulin-like growth factor binding protein-3, transforming growth factor beta, soluble Fas, and superoxide dismutase activity for the JACC study. AB - BACKGROUND: Subjects of the Japan Collaborate Cohort Study (JACC Study) gave peripheral blood samples collected between 1988 and 1990. We conducted to investigate whether levels of serum components measured after 9 years of frozen storage are stable or not. METHODS: To assess the degradation of frozen serum components in the JACC Study, we compared levels of various components (IGF-I, IGF-II, IGFBP-3, TGF-beta 1, sFas, and total SOD activity) between fresh and stored sera collected from other inhabitants. Serum levels of constituents were measured by immunoradiometric assay (IGF-I, IGF-II and IGFBP-3), quantitative enzyme immunoassay (TGF-beta1), enzyme-linked immuno-adsorbent assay (sFas), and an improved nitrite method (SOD activity). RESULTS: The coefficients of variation for intra- and inter-assay precisions of the measurements were less than 9%. Levels of IGF-I, IGF-II, IGFBP-3, TGF-beta 1 and sFas in sera after storage for 9 years at--80 degrees C were similar to those of fresh sera newly collected from inhabitants. The distributions of serum IGF-I, IGF-II, IGFBP-3, TGF-beta 1, sFas and SOD activity for specimens collected from different individuals tended to be similar to those of serum levels for frozen specimens collected from different individuals and stored for 9 years. CONCLUSIONS: There was no statistically significant difference in distribution of measured values of IGF-I, IGF-II, IGFBP 3, TGF-beta 1, and sFas between newly collected sera and frozen specimens stored for 9 years. Thus, measurements of these serum constituents of specimens stored for the JACC Study can be reliably used in nested case-control study. PMID- 15881198 TI - Mortality in the JACC study till 1999. AB - BACKGROUND: We have been conducting a cohort study named "the Japan Collaborative Cohort Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho)" since 1988. The aim of this paper is to describe the mortality of our JACC cohort in the follow up period from 1988 through 1999, to compare it with the mortality, especially cancer deaths, of the Japanese population in the same period and to compare the causes of mortality by district among the cohort. METHODS: We conducted a follow up study of 110,792 Japanese inhabitants aged 40-79 years in 1988--1990 for about 10 years to the end of 1999. RESULTS: Of 46,465 males, 37,750 (81.2%) were alive, 7,238 (15.6%) were dead and 1,477 (3.2%) had moved out of the study areas. The figures were 57,016 (88.6%), 4,940 (7.7%) and 2,371 (3.7%) among 64,327 females, respectively. The mean follow-up period was 9.9 years. The proportion of cancer deaths by site in our cohort members was almost same as the Japanese population aged 40-79 years old in 1995. Sex-specific standardized mortality ratios of total deaths, all cancer deaths, and most cancers in our cohort were less than 100 in both males and females for total cohort and the cohort by district. CONCLUSION: Our cohort members appeared to be almost the same or slightly healthier and less likely to die from total causes and cancers than the general population. PMID- 15881199 TI - Survey for incidence of cancer as a measure of outcome in the JACC study. AB - BACKGROUND: As endpoint of cohort studies on cancer, the incidence, rather than the mortality is preferable. Of 45 areas in the Japan Collaborative Cohort Study (JACC Study), surveys for incidence of cancer were conducted in 24. METHODS: The proportion of the study subjects aged 40 to 79 years in areas of survey for the incidence of cancer (65,184 persons) was 58.2% of the total subjects of the JACC Study (110,792 persons). Among the 24 areas of survey for the incidence of cancer (ASI), 10 areas were combined because of similarity. Then, we present the incidence rate of cancer among 15 ASI unified from the 24 ASI by gender according to area. We also report the completeness of the survey for cancer incidence presenting the mortality-incidence ratio (MI ratio) among the ASI. RESULTS: Where a population-based cancer registry was utilized, the MI ratio deviated from 0.31 to 0.61 in the male subjects and from 0.15 to 0.53 in the female subjects. However, where a population-based cancer registry was not used, the MI ratio deviated from 0.25 to 0.72 in the male subjects and from 0.13 to 0.79 in the female subjects, and there was an area where the MI ratio exceeded 0.70 in both of the male and female subjects. CONCLUSION: Establishment of population-based cancer registries is strongly desired throughout Japan to assess risk factors of cancer development for primary prevention of cancer. PMID- 15881200 TI - Technique-sensitivity of contemporary adhesives. AB - Besides micro-mechanical interlocking through hybrid-layer formation, self-etch adhesives may benefit from additional chemical interaction between the functional monomer and residual hydroxyapatite. One-step adhesives are commonly associated with lower bonding effectiveness, which must be attributed in part to the dissolution of hydrophilic and hydrophobic monomers in a relatively highly concentrated solvent. In this 'difficult' mixture, also water is essential as ionization medium to enable self-etching activity. Due to the high hydrophilicity, one-step self-etch adhesives have been reported to behave as semi permeable membranes, allowing fluids to pass through and seriously jeopardizing bond durability. Recent research has also revealed that HEMA-free one-step adhesives are prone to phase-separation, which may also account for their lower bonding effectiveness. Employing an appropriate air-drying technique may, however, improve the bonding effectiveness of such phase-separating adhesives by getting rid of substantially more interfacial water than HEMA-containing adhesives, of which water might be more difficult to remove from. PMID- 15881201 TI - Recycling of used commercial phosphate-bonded investments with additional mono ammonium phosphate. AB - Industrial waste is on the increase, resulting in severe environmental contamination. Against this background, this study sought to investigate the possibility of recycling used phosphate-bonded investments. Commercial phosphate bonded investment was mixed and heated according to manufacturer's instructions, then powdered with a ball mill machine to be used as recycled investment (Code: R). Mono-ammonium phosphate (NH4H2PO4) of 2, 5, 8, 11, and 14 g were added to 100 g of R because of residual MgO, and coded as Ra, Rb, Rc, Rd, and Re respectively. Investment properties were then examined. Particle size of R was 50.66 microm and close to the original investment. It was found that more NH4H2PO4 led to larger setting expansion, smaller thermal expansion, and lower fired strength. The data of Re in these three aspects were 1.42 +/- 0.09%, 0.78 +/- 0.21%, and 8.85 +/- 0.88 MPa respectively, and differences against the original investment were significant. Ra showed significantly longer setting time than other investments. Based on the results of this study, we concluded that it was possible to set used phosphate-bonded investments by adding NH4H2PO4; in particular, Rd showed suitable expansion and acceptable mechanical properties. PMID- 15881202 TI - A novel primer on dentin bonding of 4-META/MMA-TBB to collagen-depleted dentin. AB - The purpose of this study was to investigate the efficacy of an experimental dentin primer consisting of ascorbic acid (AA) and ferric chloride (Fe). Three exprimental primers - 10 wt%AA and 0.075 wt%Fe (i.e., 10AA-0.075Fe), 10AA-0Fe, and 0AA-0.075Fe - were prepared. Flattened dentin surfaces were first treated with phosphoric acid and NaClO, and subsequently primed using each experimental solution. Besides the experimental solutions, 10 wt% citric acid and 3 wt% ferric chroride (10CA-3Fe) was also used. A stainless steel rod was bonded to the dentin surface with Super-Bond C&B. 10CA-3Fe/Super-Bond C&B was then used as a control. Tensile bond strengths were evaluated after 24-hour immersion in water. 10AA 0.075Fe showed significantly high bond strength compared to 10AA-0Fe, 0AA 0.075Fe, 0AA-0Fe, and 10CA-3Fe. No significant differences were found between 10AA-0.075Fe and Super-Bond C&B system. The experimental primer consisting of 10 wt% ascorbic acid and 0.075 wt% ferric chloride improved the bonding between Super-Bond C&B and dentin conditioned with phosphoric acid and NaClO. PMID- 15881203 TI - The physical properties of a machinable resin composite for esthetic restorations. AB - To investigate the pre-clinical relevancy of a machinable composite, its physical properties were evaluated and compared with a machinable ceramic and two indirect composites. A machinable resin composite (GN-I composite, CO), a machinable ceramic (GN-I ceramic, CE), and two resin composites (Artglass dentin, AG; Estenia dentin, ET) were used. Compressive strength, diametral tensile strength, flexural strength, elastic modulus, and fracture load of standardized, premolar crown-shaped specimens were determined. In terms of compressive strength, diametral tensile strength, and flexural strength, AG showed significantly lower values than the other three materials. In terms of fracture load, specimens with 1.5 mm thick wall showed a higher value than those with 1.0 mm thick wall, and the value decreased in the order of ET, CE, CO, and AG. Marginal tipping was also observed in ET and CE. Within the limits of the current study, CO showed physical properties favorable for constructing esthetic restorations. PMID- 15881204 TI - Effect of thermal cycling on shear bond strength with different types of self etching primer for bonding orthodontic brackets using a MMA-based resin. AB - The purpose of this study was to determine the bonding durability when orthodontic bracket was bonded to human enamel using a MMA-based resin (Multibond) with two types of commercially available self-etching primer - Multibond and Megabond. Multibond primer contained a polymerization initiator, but Megabond primer did not. No significant differences in shear bond strength were observed between Multibond and Megabond primers after immersion in water at 37 degrees C for 24 hours (before thermal cycling). Multibond primer produced no significant decrease in shear bond strength after 2000 and 5000 thermo-cycles between 5 degrees C and 55 degrees C. On the contrary, Megabond primer showed significant decrease in shear bond strength after thermal cycling. The present study suggested that if Multibond resin were to be used for bonding orthodontic brackets, difference in self-etching primer's components would influence shear bond strength after thermal cycling. PMID- 15881205 TI - Effect of storage period of alginate impressions following spray with disinfectant solutions on the dimensional accuracy and deformation of stone models. AB - This study investigated the effect of storage period on dimensional change and deformation of stone models to determine an acceptable period of alginate impression storage in a sealed bag after spraying with disinfectant solution. Two alginate impression products: Aroma Fine DFIII and Alginoplast EM, characterized by greater or smaller contraction in 100% relative humidity, were used. The impressions were sprayed with 1% sodium hypochlorite or 2% glutaraldehyde solution and then stored in sealed bags. The storage periods were 1 hour, 2 hours, 3 hours, and 4 hours. The sectional profiles of the stone models obtained from each impression were measured using a three-dimensional coordinate measuring system. The results indicated that the impression characterized by greater contraction in 100% relative humidity should not be stored for even 1 hour, and that storage for up to 3 hours of the impression characterized by smaller contraction was clinically acceptable. PMID- 15881206 TI - A new laboratory polymerizing apparatus equipped with halogen and metal halide light sources. AB - We developed a new laboratory polymerizing unit (Twinkle MIII: M3) equipped with different light sources (a metal halide and two halogen lamps) that could illuminate independently. This study introduced the structure of the curing unit and examined the curing depth of a representative indirect composite (Artglass). The composite material was cured with the following modes: 1) M3 - halogen only; 2) M3 - metal halide only; 3) M3 - both halogen and metal halide (HM); 4) conventional xenon; and 5) preliminary halogen. The results obtained from analysis of variances and post-hoc tests revealed that curing depth was affected significantly (p < 0.05) by both exposure mode and time, and that HM mode exhibited the most favorable curing performance (p < 0.05) among the curing units and polymerization modes. In conclusion, the M3 unit was found to be well suited to polymerizing the composite material tested. PMID- 15881207 TI - Rare earth oxide-containing fluorescent glass filler for composite resin. AB - In recent dental care, esthetic restorative materials such as composite resin and porcelain have been widely used and studied. However, their good esthetics makes the visual inspection of restored teeth difficult. In this study, a fluorescent glass filler containing rare earth oxides - which are well-known fluorescent materials - was prepared and used in composite resin to enhance visual inspection capability with small change in color. For example, when irradiated with near ultraviolet light, an Eu2O3-containing filler fluoresced clearly and visibly. The fluorescence intensity of the prepared composite resin increased with increase of Eu2O3 content in the glass filler and with filler/resin ratio in the resin. Despite the clear fluorescence, the effect of Eu2O3 on the color change of composite resin was quite small - even when up to 10 wt% Eu2O3 was added to the glass filler. Tb4O(7-) and Dy2O3-added fillers also showed clear fluorescence, just like Eu2O3-added filler. Therefore, fluorescent glass fillers for composite resins, resulting in small color change in the latter, were successfully prepared in this study. PMID- 15881208 TI - Surface modification of pure titanium by plasma exposure and its bonding to resin. AB - The purpose of this study was to investigate the effects on shear bond strength to resin after pure titanium (Ti) was exposed to plasma under different kinds of gas atmosphere. Polished Ti samples were treated using a plasma exposure apparatus in gas atmospheres of air, CO2, and C3F8. Surface analysis of Ti exposed to plasma was achieved through surface free energy and XPS measurements. The Ti sample was bonded with adhesive resin (4-META, MAC-10, HEMA, MDP, VBATDT) to a stainless steel piece. After which, shearing adhesion test was done. It was observed that plasma exposure in a specific gas atmosphere in regulated the bonding strength of titanium surface to resin. Based on the results of this study, we concluded that plasma exposure was a useful surface treatment method for dental practices. PMID- 15881209 TI - Effect of air-powder polishing on dentin adhesion of a self-etching primer bonding system. AB - The purpose of this study was to evaluate the effect of air-powder polishing with sodium bicarbonate (SB) or crystalline cellulose (CC) on the bond strength of a self-etching primer bonding system to dentin. Ground human dentin surfaces were prepared using 600-grit SiC paper. The teeth were divided into three groups according to dentin treatment: control, air-powder polishing with SB, and air powder polishing with CC. The dentin surfaces were bonded with a self-etching primer bonding system, followed by a light-cured resin composite. microTBS test and SEM analysis were performed. The results of microTBS test indicated that air powder polishing with SB affected bond strength to dentin, while that with CC did not influence bond strength. SEM observation of air-polished dentin surfaces showed that dentin surfaces air-polished with SB were roughened and covered with a smear layer, while those with CC were smooth and the smear layer was removed. PMID- 15881210 TI - Effective factors including periodontal ligament on vertical root fractures. AB - The purpose of the present study was to analyze effective factors including periodontal ligament on vertical root fractures (VRFs). Eighty maxillary central incisors were assigned to eight groups, depending on master apical file (MAF) size, loading point and existence of a simulated periodontal ligament (PDL). The gutta-percha in the root canal was vertically pressed with a Co-Cr rod using a universal testing machine. An ordinal logistic analysis was employed to determine significant factors. VRFs were observed in 88.8% of the specimens but no horizontal fractures were found. There were three types of VRFs: partial fracture not involving the apex (PF, 41.3%), partial fracture involving the apex (AF, 40.0%), and complete fracture (CF, 7.5%). MAF size significantly affected VRFs (p < 0.05); large MAF size exhibited more partial fractures involving the apex. Loading point and existence of PDL did not affect VRFs. PMID- 15881211 TI - Influence of curing method and storage condition on microhardness of dual-cure resin cements. AB - This study evaluated the influence of curing method and storage condition on the microhardness of dual-cure resin cements: Panavia F 2.0 (PF) and Nexus 2 (NX). The specimens were either light-cured (LC) or chemically cured in darkness (CC). After 24 hours of storage in dry chamber (Dry) or distilled water (DW), the specimens were sectioned and polished. The microhardness of resin cement matrix was measured using a nanoindentation tester (ENT-1100). The data (n = 6) were statistically analyzed with t-test, two-way ANOVA (p < 0.05), and Tukey HSD test (alpha = .05). It was found that the factors of curing method and storage condition had significant effect on microhardness. For both PF and NX, LC presented higher microhardness than CC, while DW showed higher microhardness than Dry. In conclusion, dual-cure resin cements could achieve high degree of cure when light-cured. In addition, the microhardness of the resin cements evaluated did not decrease when kept in water. PMID- 15881212 TI - Fabrication of porous particulate for the scaffold by applying solution spraying method. AB - A simple and novel method--in the form of solution spraying--was developed to fabricate biodegradable, porous poly(L-lactic acid) (PLLA) particulates for scaffold. PLLA pellets were dissolved in an organic solvent. Then, 5 % PLLA dioxane solution was sprayed using an air-assisted atomizer with a nozzle diameter of 2.5 mm at an air flow rate of 15 L/min. After the sprayed solution solidified in liquid nitrogen, spherical particulates with median diameter of 225microm were obtained. Morphology of sprayed products could be altered by varying the fabrication conditions. When nozzle diameter was reduced to 1.5 mm, sprayed products became fibrous. When the concentration of PLLA-dioxane solution was increased, the diameter of particulates increased too. On the other hand, when air flow rate was increased, the diameter of particulates decreased. Likewise, solidification conditions also affected the morphology of sprayed products, such that they were either thin film-like or in particulate form. Based on the results of the present study, we concluded that PLLA particulates of varying morphologies could be obtained by adjusting the fabrication conditions. PMID- 15881213 TI - Enamel micro-cracks produced around restorations with flowable composites. AB - In this study, enamel micro-cracks produced around flowable composite restorations were observed using a stereomicroscope and a scanning confocal laser microscope (SCLM). The effects of polymerization shrinkage, mobility of composite and polishing period after filling on the incidence of marginal enamel micro cracks were examined. Enamel micro-cracks were observed on all of the composite restorations when the restoration was polished immediately after filling. Enamel micro-cracks distributed approximately parallel to the cavity margin and located 0.01-0.3 mm from the restored cavity margin. The occurrence of enamel micro cracks was higher in conventional hybrid composite restorations than in flowable composites, and when polished 15 minutes after filling (as compared to 24 hours after filling). PMID- 15881214 TI - Fluoride release/uptake of glass-ionomer cements and polyacid-modified composite resins. AB - The aim of this study was to investigate the fluoride release and fluoride recharge behaviors of two conventional glass-ionomer cements (GICs) and two polyacid-modified composite resins (PMCRs) after exposure to mouthwash and toothpaste. Fluoride released from the materials was measured at 1st, 2nd, 3rd, 4th, 7th, 14th, 21st, and 28th days. At 28th day, the specimens were divided into three groups. Specimens in the control group were stored in deionized water. For the other two groups, the specimens were exposed to 0.05% NaF solution and fluoridated toothpaste for one month. After refluoridation, fluoride amount was measured at 30th, 31st, 32nd, 36th, 44th, 52nd, and 60th days. All materials released fluoride. The highest amount of fluoride was obtained during the first 24 hours, and there was a statistically significant difference between the amounts of fluoride released from GICs and PMCRs (p < 0.0001). After exposure to mouthwash and fluoridated toothpaste, all materials were recharged and continued releasing fluoride. While the amount of fluoride release from the materials increased after reflouridation, the increase was higher in GICs. PMID- 15881215 TI - Mechanical and fluoride release properties of titanium tetrafluoride-added glass ionomer cement. AB - The aim of this study was to determine the fluoride-release and mechanical properties of a water-hardening glass-ionomer cement (GIC) (ChemFil Superior) when titanium tetrafluoride (TiF4) was added. Three experimental groups were prepared with TiF4 added to the liquid component of the material in concentrations of 0.5, 1, and 2%. The control group was the original form of the cement and free of TiF4. After the specimens (4 mm in diameter x 6 mm in length) were prepared, their compressive strength, microhardness, modulus of elasticity, and fluoride release were measured. Data were analyzed using one-way analysis of variance (ANOVA) and post-hoc test (Bonferroni/Dunn correction). The addition of TiF4 into GIC significantly reduced fluoride release from the material with the exception of 1% TiF4 (p < 0.0083). Compressive strengths of 0.5 and 1% TiF4-added GICs were higher than that of the original GIC, but it was not statistically significant (p > 0.05). The differences among modulus of elasticity values of experimental and control groups were not significant (p > 0.05). Similarly, microhardness of GIC was not affected with TiF4 addition (p > 0.05). PMID- 15881216 TI - Strengthening of calcium phosphate cement by compounding calcium carbonate whiskers. AB - The purpose of this study was to investigate how aragonite (calcium carbonate) whiskers influenced the strengthening and carbonating of alpha-tricalcium phosphate (alpha-TCP) based calcium phosphate cement. Aragonite whiskers of 0.95 microm width with an aspect ratio of 6.6 were prepared. The cement powder, alpha TCP containing 0-50 mass% aragonite whisker, was mixed with 0.6 mol/L NaH2PO4 solution and incubated at 37 degrees C and 100% relative humidity. Diametral tensile strength (DTS) value increased significantly when appropriate amount of aragonaite whiskers was added. For example, DTS value of set cement containing 20 mass% aragonite whisker was 5.8 +/- 0.5 MPa, whereas DTS value of set cement containing no whiskers was 1.3 +/- 0.2 MPa after 1-week incubation. SEM observation revealed that the shape of the whiskers and the densification of the structure could have contributed to the strengthening of the set cement. Moreover, FTIR spectra implied that a bone-like carbonated apatite was precipitated in the cement. The results obtained in the present study revealed that the shape as well as any slight dissolution of aragonite whiskers could contribute to improving the properties of a-TCP based calcium phosphate cement. PMID- 15881217 TI - Translucency and characteristics of newly developed polymer-based dental tooth coating material. AB - The purpose of this study was to evaluate the translucency and color change of simulated heavily discolored teeth using polymer-based dental tooth coating materials and flowable resin composites. Five shades of coating material and two shades of flowable resin composite were used. Colorimetric values of the materials in different shades were determined using the L*a*b* system of the Commission Internationale de l'Eclairage (CIE). Colorimetric examination was performed on white, black, and shade guide (C4) backgrounds using specimens of various thickness (0.2, 0.3, 0.5, 1.0, and 2.0 mm). New coating material showed less translucent than flowable resin composites. Moreover the new material showed the potential to improve the appearance of heavily discolored teeth when being applied as a thin first layer. PMID- 15881218 TI - Effect of resin coating as a means of preventing marginal leakage beneath full cast crowns. AB - The purpose of this study was to evaluate the effectiveness of resin coating as a means of preventing marginal leakage beneath full cast crowns which were emplaced using different cements. Standard full cast crown preparation was made on 64 extracted premolars. These samples were then divided into four groups, with half of each group coated with dentin coating material after preparation. Crowns were cemented onto the teeth using zinc cement, Fuji I, Vitremer, or C&B Metabond. The samples were thermal-cycled for 10,000 cycles. They were then immersed in erythrosine solution, sectioned, and observed under a microscope. Microleakage analyses were performed using a 0-4 point system. The data were statistically analyzed. There were significant differences between the coated specimens and the uncoated specimens using Fuji I and Vitremer. The results showed that a resin coating could decrease the amount of marginal leakage when applied with these two cements. PMID- 15881219 TI - Machining accuracy of CAD/CAM ceramic crowns fabricated with repeated machining using the same diamond bur. AB - The purpose of this study was to investigate the effect of repeated machining up to 51 times using the same diamond bur on machining accuracy of inner and outer surfaces of CAD/CAM (computer-aided designing and computer-aided manufacturing) machined ceramic crowns. The surface topography of machined crowns was examined using photographs. It was found that machining accuracy was not affected by the number of machining times. In all measuring points, the inner surface was machined to a dimension larger than the die model (i.e., increased gap), whereas the outer surface was machined to a dimension smaller than the crown model (i.e., smaller crown). Photo observation showed that cervical contour was machined in a clear, rounded form from 1st to 11th crowns. PMID- 15881220 TI - Durability of diamond burs for the fabrication of ceramic crowns using dental CAD/CAM. AB - The diamond burs of two dental CAD/CAM systems (GN and CD) were examined if they could be used to fabricate up to 21 ceramic full crowns without fracture. After one, 11, and 21 machining times, the surfaces of the diamond burs were observed and the number of particles captured on SEM pictures was counted. The average surface roughness of the crowns was also measured. All diamond burs could be used to fabricate 21 ceramic crowns without fracture. A significant decrease in the number of diamond particles was found on the surfaces of GN burs after 11 and 21 machining times, but not on those of CD burs. The average surface roughness of GN crowns significantly increased with increase in the number of machining times. A significant positive correlation was found between the average surface roughness and the number of diamond particles. PMID- 15881221 TI - High temperature characteristics and solidification microstructures of dental metallic materials. Part III alloys for metal-bond porcelain. AB - The thermal expansion rate, coefficient of thermal expansion, and high temperature strength of two types of commercially available alloy for metal-bond porcelain, KIK-HII (KIK) and Degubond-J2 (J2), were evaluated up to the liquidus point temperature using a thermo-mechanical analyzer. Furthermore, microstructure in the solid-liquid coexisting region was observed for evaluation. Our results revealed the following findings: 1. For KIK, solidus point was 1,209.3 +/- 3.2 degrees C, liquidus point was 1,308.3 +/- 7.10 degrees C, and melting expansion rate was 0.41+/- 0.16%. 2. For J2, solidus point was 1,198.3 +/- 0.6 degrees C, liquidus point was 1,253.0 +/- 4.4 degrees C, and melting expansion rate was 4.50 +/- 0.80%. 3. At high temperature, the mechanical characteristics of KIK greatly differed from those of J2. The risk of causing deformation during porcelain baking was suggested for KIK. Removal of segregation during casting was considered difficult in J2. PMID- 15881222 TI - A comparative study of oral health attitudes and behaviour using the Hiroshima University - Dental Behavioural Inventory (HU-DBI) between dental students in Britain and China. AB - The aim of this study was to compare oral health attitudes and behaviour between British and Chinese dental students. A cross-national survey using the Hiroshima University - Dental Behavioural Inventory (HU-DBI) was completed at the University of Leeds in Britain and West China University of Medical Sciences in China. In Britain and China, 192 of 266 and 180 of 303 dental students answered English and Chinese versions of the HU-DBI questionnaire, respectively. Data were statistically analyzed by logistic regression and the following results were obtained: 1) Self-reported gingival bleeding was more prevalent in Chinese students than in British students, although the number of students who had professional oral hygiene instruction was higher in China than in Britain; 2) 29% of Chinese students believed wearing dentures in old age was inevitable, whereas 7% of British students believed so; 3) Chinese students were substantially more concerned about the appearance of their teeth, gums and halitosis; and 4) 54% of Chinese students seek dental care only when symptoms arise, as compared to 13% of British students. In conclusion, bilateral comparison of countries using HU-DBI revealed interesting differences in oral health attitudes and behaviour, while the logistic regression model made it possible to differentiate British from Chinese students with a probability of nearly 95%. PMID- 15881223 TI - Influence of temporary cement remnant and surface cleaning method on bond strength to dentin of a composite luting system. AB - The aim of the current study was to evaluate the influence of polycarboxylate temporary cement remaining on the dentin surface on the bond strength of a composite luting system. An acrylic resin plate was luted to bovine dentin with a polycarboxylate temporary cement (HY-Bond Temporary Cement Hard, HYB). The temporary cement was not used for the control groups. After removing the temporary cement with an excavator, dentin specimens were divided into five groups; 1) no subsequent treatment, 2) cleaning with a rotational brush (RTB), 3) cleaning with a rotational brush and non-fluoridated flour of pumice, 4) sweeping with an air scaler, and 5) treated with a sonic toothbrush. A silane-treated ceramic disk (IPS Empress) was bonded to each dentin specimen with a composite luting system (Panavia F). Shear testing results showed that the RTB groups exhibited the highest bond strength regardless of the use of temporary cement (P < 0.05). The use of a rotational brush with water coolant is recommended to achieve ideal bond strength between the Panavia F luting system and dentin to which HYB temporary cement was primarily applied. PMID- 15881224 TI - Transient expression of polymeric immunoglobulin receptor in human adenocarcinoma cell line HT-29. AB - Human polymeric immunoglobulin receptor (pIgR) protein was expressed in the adeno carcinoma cell line HT-29 using a recombinant vaccinia virus transfection method. The pIgR protein was detected as 110- and 120-kDa bands by immunoprecipitation after metabolic labeling. PIgR was released as a free secretory component into the culture supernatant and was detected as a 110-kDa band. PIgR cleavage was investigated by adding the proteinase inhibitor leupeptin or protein kinase C activator PMA. Consistent with previous observations in the Madin Darby canine kidney cell system, cleavage of pIgR was inhibited by leupeptin and enhanced by PMA stimulation, thus indicating that it is regulated by common mechanisms. This experimental system should be very useful for pIgR investigation. PMID- 15881225 TI - Influence of polishing duration on surface roughness of resin composites. AB - There is a need for effective polishing instruments for resin composite restorations with highly polished surfaces. The purpose of this study was to investigate the influence of polishing duration on surface roughness of light cured resin composites. Four polishing systems, Compomaster (Shofu), Silicone Points C Type (Shofu), Super Snap (Shofu) and Enhance Finishing and Polishing System (Dentsply/Caulk), were used to polish two commercially available resin composites, Clearfil AP-X (Kuraray Medical) and Lite-Fil II A (Shofu). Resin pastes were condensed into molds (10 mm in diameter, 5 mm in depth) and light irradiated for 40 s. Composite surfaces were ground with # 600 SiC paper followed by polishing with an instrument for 30 s, and the surface roughness was measured every 10 s during polishing procedures. The average surface roughnesses (Ra) were determined using a profilometer. Data were analyzed by Tukey HSD test (P = 0.05). After 30 s of polishing, mean Ra values ranged from 0.07 to 0.50 for Clearfil AP X, and from 0.11 to 0.57 for Lite-Fil II A. Although the time required for polishing was longer, the surface finish produced by multiple-step polishing systems was superior to that obtained with one-step polishing systems. PMID- 15881226 TI - Ameloblastin and amelogenin expression in posnatal developing mouse molars. AB - Ameloblastin and amelogenin are structural proteins present in the enamel matrix of developing teeth. Here we report the results of in situ hybridization analyses with DNA probes of ameloblastin and amelogenin expression in the mandibular first molars of ICR/Jcl mice from postnatal day 1 to day 15. Ameloblastin mRNA expression was observed in ameloblasts at day 2 while amelogenin mRNA was detected in secretory ameloblasts at day 3. Significant expression of both molecules was observed at days 4, 5 and 6, after which the levels decreased. Amelogenin expression ended on day 10, while ameloblastin mRNA was only weakly detected on day 12. Neither amelogenin nor ameloblastin expression was observed in day 15 mouse molars. Amelogenin and ameloblastin mRNAs were restricted to ameloblasts. We conclude that amelogenin and ameloblastin expression is enamel specific, and suggest that these genes might be involved in the mineralization of enamel. It is possible that ameloblastin could participate in the attachment of ameloblasts to the enamel surface. In this case, the downregulation of expression may indicate the beginning of the maturation stage in which the ameloblasts tend to detach from the enamel layer. PMID- 15881227 TI - Fluoride-releasing capacity and cariostatic effect provided by sealants. AB - This study evaluated the effect of sealants on enamel demineralization, focusing on physical protection of the sealed enamel and fluoride protection of the adjacent unsealed enamel. Occlusal fissures with areas measuring 12 mm2 were delimited in 48 extracted molars, randomly divided into 4 groups (n =12): 1) no sealing; 2) sealing with a resin-modified glass-ionomer (Vitremer, 3M ESPE); 3) sealing with a fluoride-releasing composite sealant (Clinpro Sealant, 3M ESPE); and 4) sealing with a non-fluoridated composite sealant (Concise, 3M ESPE). A 4 mm2 window was outlined on the buccal enamel for analysis of fluoride uptake. Following treatment, groups 2, 3 and 4 were subjected to 5-days of pH-cycling, while group 1 was kept in a moist environment at 37 degrees C. Fluoride uptake was assessed by dental biopsy, and the amount of fluoride released to the cycling solutions was determined by ion analysis. Enamel demineralization around the sealants was evaluated by cross-sectional micro-hardness analysis. Group 2 showed higher levels of fluoride release (P < 0.01) and uptake by enamel (P < 0.05), and lower levels of demineralization (P < 0.05) than groups 3 and 4. Group 3 exhibited reduced demineralization on unsealed enamel and provided fluoride uptake in a distant enamel area, while group 4 did not. PMID- 15881228 TI - Sphingosine 1-phosphate acts as a signal molecule in ceramide signal transduction of TNF-alpha-induced activator protein-1 in osteoblastic cell line MC3T3-E1 cells. AB - We previously demonstrated that tumor necrosis factor (TNF)-alpha stimulated the production of activation protein (AP)-1, a transcriptional factor, in mouse osteoblastic MC3T3-E1 cells. Recent studies have shown the importance of ceramide and its metabolites as signal molecules for TNF-alpha-induced gene expression in several cell types. Therefore, our interest was to investigate whether sphingosine metabolites are involved in TNF-alpha-induced signaling in MC3T3-E1 cells. DL-threo-1-phenyl-2-hexadecanoyl-amino-3-pyrrolidino-1-propanol (PPPP), which causes accumulation of intracellular ceramide, stimulated the TNF-alpha induced expression of the c-fos and c-jun genes. Gel shift assay clearly showed that PPPP increased the cytokine-induced specific binding of nuclear proteins to the 12-tetra-decanoyl phorbol 13-acetate-responsive element (TRE), a consensus sequence for AP-1. In addition, cell-permeable ceramide (N-acetylsphingosine, N hexanoylsphingosine or N-octanoylsphingosine) stimulated expression of the c-fos and c-jun genes and nuclear protein binding to TRE. Interestingly, DL-threo dihydrosphingosine (DHS), an inhibitor of sphingosine kinase, clearly blocked the ceramide analogue-induced stimulation. Sphingosine 1-phosphate (SPP) actually induced expression of these oncogenes and activated AP-1. Although TNF-alpha stimulated the AP-1-mediated expression of the monocyte chemoattractant JE/MCP-1, this stimulation was inhibited by DHS. SPP also stimulated JE/MCP-1 gene expression. The present study thus suggests that SPP acts as a signal molecule in ceramide-dependent signal transduction in TNF-alpha-induced AP-1 in osteoblastic MC3T3-E1 cells. PMID- 15881229 TI - A marker of oxidative stress in saliva: association with periodontally-involved teeth of a hopeless prognosis. AB - The aim of this study was to determine the association between levels of a marker of oxidative stress, 8-hydroxydeoxyguanosine (8-OHdG), in saliva and the presence of teeth with a hopeless prognosis as a result of advanced periodontitis. Thirty four periodontitis patients were divided into two groups based on the presence or absence of periodontally-involved teeth of hopeless prognosis. Salivary levels of 8-OHdG in those with were significantly higher than in subjects without periodontally-involved teeth of hopeless prognosis (4.78 +/- 0.14 ng/ml and 2.35 +/- 0.18 ng/ml, respectively). We also evaluated 8-OHdG levels in gingival crevicular fluid (GCF) of teeth with advanced periodontal destruction (mean probing depth = 7.2). In this case, 8-OHdG was detected only from those periodontally-involved teeth of hopeless prognosis, and only in some cases (8 out of 18 samples). These data suggest that periodontally-involved teeth of hopeless prognosis are a major source of salivary 8-OHdG. Measurement of salivary 8-OHdG levels may prove to be useful in identifying patients with teeth of hopeless prognosis. PMID- 15881230 TI - [Strategy for thrombosed type A acute aortic dissection]. AB - The surgical treatment for thrombosed type A aortic dissection is controversial. We treated 15 cases (28.8%) with thrombosed type A acute aortic dissection among 52 cases of all type A acute aortic dissection during past 7 years and 8 months. Nine cases was treated conservatively and 6 cases received emergent operation due to cardiac tamponade, shock, and ulcer-like projection (ULP) in the ascending aorta (> 50 mm). In the group of conservative therapy, 2 cases in which the ascending aorta was enlarged (54, 60 mm), was converted to operation in the acute phase because of further enlargement of ascending aorta and increase of cardiac effusion. The other patients (aortic diameter: 40-46 mm) who received conservative therapy were well controlled by medical therapy in the early and late phase except 1 who died suddenly in the follow-up. There was no mortality and late death in the operated patients. From these results, we conclude that thrombosed type A aortic dissection in which the diameter of ascending aortic aorta is larger than 50 mm, have cardiac tamponade and ULP in the ascending aorta might needs operation. PMID- 15881231 TI - [Acute type A aortic dissection associated with intestinal necrosis]. AB - We report a case with acute type A aortic dissection associated with occlusion of the celiac artery (CA) and the superior mesenteric artery (SMA) who underwent replacement of the aortic valve and the ascending aorta concomitant with CA and SMA bypass grafting and partial resection of the ileum and the cecum. A 79-year old man who has a history of Y-graft replacement of abdominal aortic aneurysm developed re-dissection of thrombosed type A acute dissection 2 weeks after the onset. We performed CA and SMA bypass grafting followed by replacement of the aortic valve and the ascending aorta. In spite of successful revascularization of the CA and SMA, residual necrosis in the ileum and the cecum necessitated resection of them. He was discharged 11 months postoperatively without neurological sequelae. PMID- 15881232 TI - [Completion pneumonectomy 9 years after middle lobectomy for adenocarcinoma]. AB - Completion pneumonectomy (CP) is a difficult operation in which the surgeon must use techniques such as intrapericardial ligation of the pulmonary vessels. We report herein a case of CP for a patient with recurrent lung cancer. A 63-year old man was admitted to our hospital for evaluation of abnormal shadows in the right lung field in October 2002. Right middle lobectomy with mediastinal lymph node dissection had been performed in February 1993. Computed tomography (CT) revealed a hilar mass in the right upper lobe the day after admission. Bronchofiberscopic cytology revealed squamous cell carcinoma. Right completion pneumonectomy was performed on suspicion of metachronous multiple lung cancers 4 days later. Histopathologically, resected specimens represented adenosquamous carcinoma similar to the prior lesion from the middle lobe, and examination revealed that the tumor represented a recurrence following middle lobectomy. The patient remains well as of 19 months postoperatively. PMID- 15881233 TI - [Aortic dissection induced by symmetry aortic connector system]. AB - A 74-year-old male with severe triple vessel disease underwent off-pump coronary artery bypass grafting (OPCAB). Preoperative computed tomography (CT) showed severely calcified ascending aorta. We revasculize the left coronary arteries with in situ internal thoracic artery (ITA) graft and the right coronary artery with a saphenous vein graft, which was attached to the disease-free portion of the aortic root, using Symmetry aortic connector system (ACS). Although the operation was uncomplicated, and postoperative course was uneventful until the 5th postoperative day when acute type A aortic dissection occurred. The patient died of aortic rupture on the 7th postoperative day. Necropsy disclosed that the entry located just on the proximal anastomotic site of the vein graft. It is possible placement of ACS device would trigger the dissecting process. With regard to the use of these one-shot devices for diseased aorta, its safety needs further investigation, even though it might be placed on an apparently intact portion. PMID- 15881234 TI - [Assessment of English literatures evidence toward the development of clinical guidelines for elderly patients with lung cancer]. AB - The purpose of this study is that we propose the clinical practice guidelines for elderly patients with lung cancer in Japan. The results and conclusions are as follows: 1) As pneumonectomy in patients over the age of 70 carries a considerable risk of severe postoperative morbidity and mortality, it should be avoided. 2) Although it is no significant differences between 3 groups (age < 70, 71-79, 80 < or =) in the 30 days mortality rates, less invasive surgery decrease them. 3) Age is not the important prognostic factor, however, operative procedure should be choose over the estimation of surgical risk and the potential long-term benefit. PMID- 15881235 TI - [The relationship between supraventricular arrhythmia after coronary artery bypass grafting and autonomic nerve system]. AB - We evaluated the relationship between supraventricular arrhythmia after coronary artery bypass grafting (CABG) and autonomic nerve system. Holter electrocardiogram was recorded in 57 postoperative patients. The heart rate variability was analyzed in 25 premature atrial contraction (PAC) cases and 4 atrial fibrillation (AF) cases. Sixty minutes just before the onset of PAC/AF were devided into 12 equal parts, and NN 50, RMSSD, HF, LF/HF were measured respectively. NN 50, RMSSD and HF were each used as an indicator to show the parasympathetic nerve (PSN) activity, and LF/HF as an indicator to show the sympathetic nerve (SN) activity. The mean value of the former 6 parts was compared to that of the latter 6 parts. As for 25 PAC cases, PAC occurred under the dominance of PSN in 30% of the cases, under the dominance of SN in 10% of the cases and under the balanced condition of both nerves in 60% of the cases. As for 4 AF cases, AF occurred under the parasympathetic condition in 3 cases and under the sympathetic condition in 1 case. These results show that supraventricular arrhythmia after CABG is associated with PSN system. PMID- 15881236 TI - [Surgical technique of leaflet extension in aortic valve regurgitation]. AB - Detailed procedure of the leaflet extension technique in aortic valve regurgitation is described. The concept of the technique is to create a wide and deep coaptation zone by extending the prolapsed leaflets using glutaraldehyde preserved autologous pericardium, and by suspending the commissures. The leaflet extension technique is a procedure of choice for aortic valve regurgitation in young patients who need to avoid strict anticoagulation therapy. A careful long term follow-up is necessary to evaluate the durability of glutaraldehyde preserved pericardium in aortic position. PMID- 15881237 TI - [Measurement of air leak volume after lung surgery using web-camera]. AB - Persistent air leak from the lung is one of the major complications after lung operations, especially in the latest thoracic surgery, where a shorter hospital stay tends to be necessary. However, air leak volume has been rarely measured clinically because accustomed tools of gas flow meter were types which needed contact measure, and those were unstable in long-term use and high cost. We tried to measure air leak volume as follows: (1) Bubble was made in the water seal part of a drain bag. (2) The movement of bubbles was recorded with a web-camera. (3) The data from the movie was analyzed by Linux computer on-line. We believe this method is clinically applicable as a routine work after lung surgery because of non-contact type of measurements, its stableness in long-term, easiness to be handled, and reasonable in cost. PMID- 15881238 TI - [Thoracoscopic resection for Mycobacterium avium complex]. AB - We report 2 cases of thoracoscopic resection for patients with Mycobacterium avium complex (MAC). A 25-year-old female was referred to our hospital because of abnormal shadows in the right lower lung field on chest X-ray. Her chest computed tomography (CT) showed that the lesion was localized in the right lower lobe. She was given a diagnosis of MAC by polymerase chain reaction (PCR) of sputum. Thoracoscopic lobectomy was performed after chemotherapy for 10 months. A 64-year old female was referred to our hospital because of hemoptysis. Bronchiectasis had been diagnosed in her since the age of 35 years, and then she was given a diagnosis of secondary MAC. Her chest CT showed bronchiectasis, and consolidations were localized in the superior segment of the right lower lobe. Then superior segmentectomy of the right lower lobe under video-assisted thoracoscopic surgery (VATS) was performed. It was reported that surgical intervention is indicated in patients with MAC, when persistent hemoptysis is seen or chemotherapy is ineffective. If the lesion is localized, lung resection under VATS may be a good option in selected patients. PMID- 15881239 TI - [Primary cardiac angiosarcoma in the left atrium with adrenal metastasis; report of a case]. AB - We encountered a 61-year-old woman with primary cardiac angiosarcoma in the left atrium. On echocardiography, the tumor extended into the atrial septum and mitral valve, and mitral valve stenosis and regurgitation were significant. We resected the tumor protruding into left atrium, and affecting mitral valve. The surgical procedure was not radical, but on postoperative echocardiography, function of the mitral valve was improved. Three months later, elevation of her right diaphragma was observed on chest X-ray and a giant adrenal tumor was detected by magnetic resonance imaging. Tumor biopsy indicated that this tumor was adrenal metastasis from cardiac angiosarcoma. In addition, echocardiography showed the recurrence of angiosarcoma in the left atrium and the presence of mitral stenosis and regurgitation. She died of heart failure 185 days postoperatively. PMID- 15881240 TI - [Lung cancer which accompanies anomalous venous connections, pulmonary and systemic-partial anomalous pulmonary venous connection and persistent left superior vena cava: report of 2 cases]. AB - We report the cases of a left partial anomalous pulmonary venous connection (PAPVC) and a persistent left superior vena cava (PLSVC), combined with primary lung cancer. Our case of PAPVC, the anomalous pulmonary vein originated from the hilum of the left upper lobe flowed into the left brachiocephalic vein. A left lower lobectomy was performed uneventfully without correcting the anomalous vein. And a case of PLSVC, the left superior vena cava flowed into the right superior vena cava, running under the aortic arch. A left upper lobectomy and mediastinal lymph node dissection was performed in safety. Although PLSVC was detected by chest computed tomography (CT) before operation, PAPVC was noticed intraoperatively in our case. We should keep in mind the possibility of variations of pulmonary vessel distribution, especially PAPVC located in a different lobe for resection, when undertaking lung resection. PMID- 15881241 TI - [Video-assisted thoracoscopic surgery for migration of an orthopedic fixation wire in the mediastinum: report of a case]. AB - Migration of an orthopedic fixation wire into the thoracic cavity is an uncommon complication. We present a 64-year-old male undergoing intra-thoracic migration of a Kirschner wire. The patient had undergone the treatment of sternoclavicular joint dislocation due to clavicular fracture. Four months after the operation, a chest X-ray revealed a metallic foreign body in the mediastinum. A chest computed tomography (CT) showed the object in the anterior mediastinum adjacent to the aortic arch. An emergency video-assisted thoracoscopic surgery was successfully performed to remove a 60-mm-long Kirschner wire migrated from the mediastinum to the thoracic cavity. The operating time was 45 minutes, and the blood loss was less than 50 ml. His postoperative course was uneventful and he was discharged at the 6th day after surgery. Prompt diagnosis and surgical removal are necessary to prevent serious complications. PMID- 15881242 TI - [Successful removal of an intrapulmonary aberrant needle under video-assisted thoracoscopic surgery; report of a case]. AB - Intrapulmonary aberrant needles are rare in clinical practice. We report the successful removal of intrapulmonary aberrant needle. A 59-year-old man, though he was asymptomatic, was referred to our department after an abnormal shadow had been detected on a chest X-ray. Chest X-ray and chest computed tomography (CT) showed a foreign body suspected to be a metal artifact in the left upper lobe. It was diagnosed as an intrapulmonary aberrant needle and an operation under video assisted thoracoscopic surgery was performed. Using perioperative fluoroscopy, we could confirm the location of the needle and remove it successfully. An intrapulmonary aberrant needle should be removed surgically, even if the patient is asymptomatic, due to the development of lung abscess or pyothorax and the risk containing harmful matter to health. PMID- 15881243 TI - [Gingival metastasis of pulmonary pleomorphic carcinoma; report of a case]. AB - A 71-year-old male was admitted to the hospital complaining of cough. The chest X ray and computed tomography (CT) revealed a large tumor in the right lower lung, which was diagnosed as poorly differentiated adenocarcinoma. As the tumor grew rapidly and caused obstructive pneumonia, right middle and lower lobectomy was performed even if right gingival tumor was suspected as metastasis from lung tumor. The patient complicated with aspiration pneumonia after operation and died on the 20th postoperative day. The prognosis of lung cancer with gingival metastasis is very poor. Early detection and appropriate therapy is necessary. PMID- 15881244 TI - [Pyopneumopericardium caused by perforation of gastric ulcer; report of a case]. AB - Pyopneumopericardium caused by transdiaphragmatic perforation of gastric ulcer is very rare. A 60-year-old man was admitted to our hospital because of chest pain with high fever. The chest computed tomography (CT) revealed hydropneumopericardium. The patient was diagnosed as purulent pericarditis. So emergent pericardiectomy and pericardial drainage were performed to relieve cardiac tamponade. Two findings enabled us to diagnose the causation of pyopneumopericardium as gastropericardial fistura. The first finding was that endoscopic examination of upper gastrointestinal tract revealed a deep peptic ulcer in the dome of gastric fundus. The second finding was that a dye solution which was injected into the pericardial cavity via the drainage tube leaked out into the gastric cavity through the ulcer. This patient improved successfully by the treatment of intravenous hyperalimentation including antibiotics and omeprazole. We think that expedient diagnosis and surgical drainage are essential for successful patient outcome. PMID- 15881245 TI - [Thoracoscopy and intraoperative upper gastrointestinal endoscopy was effective for Boerhaave syndrome; report of a case]. AB - Boerhaave syndrome is a rare disease and needs an exact diagnosis and a proper treatment plan because of its terrible clinical course. We experienced a case of Boerhaave syndrome that thoracoscopy and intraoperative upper gastrointestinal (GI) endoscopy was very effective. Sixty-four-year-old man realized chest and back pain after vomitting. Esophageal perforation was suspected, but 64 hours had passed already when we started a surgical treatment. By the thoracoscopy and intraoperative endoscopy, lower esophageal perforation and infectious pleural effusion were found. Therefore, we selected a surgical treatment under the assistance of thoracoscopy. Secondly, a simple closure and intracostal muscle overlapping was performed with small incisional thoracotomy. Postoperative complication, such as mediastinal abscess, has not occurred. Thoracoscopy and intraoperative upper GI endoscopy was effective for an appropriate diagnosis and treatment of Boerhaave syndrome. PMID- 15881246 TI - [Hemothorax following intraabdominal bleeding due to pedicle torsion of ovarian tumor via a small pore in the diaphragm: report of a case]. AB - On February 25, 2004, 59-year-old woman visited a local clinic due to lower abdominal pain. On February 28, she was admitted to the clinic due to severe abdominal pain. Computed tomography (CT) showed a mass in the lower abdomen and plural effusion and athelectasis of the right lung. She had severe anemia (Hb 6.9 g/dl). On March 1, she was transferred to our hospital. Pleural fluid was revealed to be sanguineous by thoracentasis. She underwent thoracotomy on the day of admission. There was no source of bleeding in the pleural space. A small pore, 3 mm in diameter, was found in the tendinous portion of the diaphragm. An influx of bloody fluid from the abdomen via the pore caused hemothorax. Laparotomy was performed, followed by closure of the pore using direct suture. The origin of the abdominal bleeding was pedicle torsion of the right ovarian tumor. Seven months after surgery she was uneventful with no pleural effusion. PMID- 15881247 TI - [Diaphragmatic eventration resulting from phrenicectomy treated with surgical method; report of a case]. AB - The case was a 59-year-old man who has a history of left mediastinal tumor resection with left phrenicectomy. The elevated diaphragm revealed by chest X-ray 7 years after the operation led to diagnosis of diaphragmatic eventration. Since any symptom was seen in the early period, "wait and watch" strategy was done for management. Both the abdominal enlarged feeling and the dyspnea on effort were appeared 10 years after the operation. Under the speculation of these symptoms related to the elevated abdominal organs came up with diaphragmatic eventration, surgical method the plication of the diaphragm was performed. The diaphragm was plicated by interrupted suture as opening the diaphragm to avoid injury the abdominal organs, and reinforced with the Marlex mesh. We used artificial mesh to reinforce the thin diaphragm with exceptation of prevent the postoperative recurrence, because a result of the etiological process of the case was considered as disuse atrophy of diaphragm after phrenicectomy. PMID- 15881248 TI - [Heat treatment for the control of Bacillus cereus spores in foods]. AB - The effect of heat treatments on food products related to soybean curd contaminated with Bacillus cereus spores was investigated for the purpose of preventing food-poisoning outbreaks by B. cereus. In the case of B. cereus strains isolated from foods, heating foods inoculated with the spores for 20 min at 70 degrees C, 5 min at 75 degrees C, 2 min at 100 degrees C, or 10 sec with a microwave oven plus reheating after standing for 2 hr at 25 degrees C reduced the number of surviving cells within the foods to less than one-hundredth. In the case of a heat-resistant strain isolated from a food poisoning outbreak, heating for 20 min at 70 degrees C plus reheating after standing for 1.5-2 hr at 35 degrees C, heating for 10 min at 75 degrees C plus reheating after standing for 1.5-2 hr at 35 degrees C, or heating for 2 min at 100 degrees C plus reheating after standing for 4 hr at 25 degrees C was effective. There was not much difference between the flavor components in foods with and without heat treatment at 70 degrees C, as analyzed by gas chromatography-mass spectrometry. These results indicate that these heat treatments are available to control B. cereus spores, without affecting the sensory quality of the foods. PMID- 15881249 TI - [Headspace GC/MS analysis of residual vinyl chloride and vinylidene chloride in polyvinyl chloride and polyvinylidene chloride products]. AB - A headspace GC/MS analysis method for the simultaneous determination of residual vinyl chloride (VC) and vinylidene chloride (VDC) in polyvinyl chloride (PVC) and polyvinylidene chloride (PVDC) products was developed. A test sample was swelled overnight with N,N-dimethylacetamide in a sealed vial. The vial was incubated for 1 hour at 90 degrees C, then the headspace gas was analyzed by GC/MS using a PLOT capillary column. The recoveries from spiked PVC and PVDC samples were 90.0 112.3% for VC and 85.2-108.3% for VDC. The determination limits were 0.01 microg/g for VC and 0.06/microg/g for VDC, respectively. By this method, VC was detected in two PVC water supply pipes at the levels of 0.61 and 0.01 microg/g. On the other hand, VC and VDC were not detected in any of the food container packages or toys tested. PMID- 15881250 TI - [Analysis of residual volatiles in recycled polyethylene terephthalate]. AB - The residual volatiles in recycled polyethylene terephthalate (PET) were analyzed using headspace/GC/MS. Recycled PET samples were made from PET bottles used for beverages, alcohol and soy sauce, and they were recycled in physical recycling plants, chemical recycling plants and superclean-like recycling trials. The physically recycled PET flakes contained small amounts of volatiles such as ethanol, limonene, 2-methyl-1,3-dioxolane, acetone, octanal and nonanal. Most of them originated from foods packed in bottles, and only 2-methyl-1,3-dioxolane was derived from polymer impurities. In contrast, the superclean-like or chemically recycled PET contained no detectable volatiles, like new PET pellets. The PET sheets shaped from physically recycled PET had no detectable volatiles. Not only the chemically and superclean-like recycled PET, but also the physically recycled PET contained no hazardous volatiles. It was concluded that there is no safety concern about volatiles in recycled PET, for the present use. PMID- 15881251 TI - [Laboratory-performance study of the notified methods to detect genetically modified papaya (55-1)]. AB - To investigate important factors affecting the reliability of the analytical results, proficiency tests were attempted for the histochemical method (GUS method) and the qualitative PCR method (PCR method) to detect genetically modified papaya (55-1) in the Japanease official method. The test samples were distributed to twenty-three laboratories that participated in the study and were examined according to the protocol. All the data collected from participating laboratories were statistically analyzed. In the PCR method, one negative sample was detected as positive using detection primers in one laboratory, though the sample was negative when checked using confirmation primers. Contamination might have occurred in the step of the preparation of the PCR sample solution using detection primers. In the GUS method, all the test samples were identified as expected. Thus, all the laboratories reported correct results overall. PMID- 15881252 TI - [Determination of isophorone in foods]. AB - Isophorone (ISP) is used widely as a solvent of natural and synthetic resins, wax, printing ink, pesticides and paints. In this study, the level of ISP in various foods (93 samples) was analyzed. ISP was collected from samples by steam distillation after the addition of an internal standard, deuterium-labeled ISP, then extracted with dichloromethane, cleaned up on a silica gel column, and determined by GC/MS. ISP was barely detected in fish, meat and vegetable samples, but it was detected in rice, wheat, beans and their processed products, miso, soy sauce and fermented soybeans (natto). The maximum level was 8.9 ng/g in miso. The packaging materials of the foods contained little ISP, and so the source of ISP in the foods could not be clarified. PMID- 15881253 TI - [The present condition and a view of the food inspection using the invader method]. PMID- 15881254 TI - Current status of regulation on GM food in Korea. PMID- 15881255 TI - A fast system for reporting the Farnsworth-Munsell 100-hue colour vision test. AB - We have developed a system for rapidly reporting the Farnsworth-Munsell (FM) 100 hue test using a personal computer and a bar code scanner. The computer generated report duplicates the conventional manual report of the FM 100-hue test so is very familiar to ophthalmologists and optometrists. The new system has proved to be of great assistance both in saving time and in eliminating arithmetic errors in the scoring calculations. The scanner technique produces two reports, one for each eye, within 4 min of the patient completing the test. This compares with the 60 min required by the conventional manual reporting system. In addition, it also gives a statistical analysis of the results in accordance with Verriest norms. The program is very versatile and user friendly, achieving a standard not present in the other FM 100-hue computerised systems currently available. As a consequence it makes this valuable diagnostic test much more accessible to patients and clinicians. PMID- 15881256 TI - Macular function assessed with mfERG before and after panretinal photocoagulation in patients with proliferative diabetic retinopathy. AB - PURPOSE: To examine macular function and its correlation to macular thickness before and after panretinal photocoagulation for proliferative retinopathy in diabetic patients. METHODS: Ten diabetic patients (aged 57 +/- 10 years, diabetes duration 21 +/- 10 years) treated with panretinal photocoagulation outside the great vascular arcade underwent multifocal electroretinography (mfERG) and optical coherence tomography (OCT) before and 6 months after treatment. When focal treatment in the macular region was performed prior to panretinal photocoagulation the investigations took place 3 weeks after this treatment but before the panretinal photocoagulation. One eye per patient was examined. Amplitudes and implicit times of the mfERG response were analyzed within the four innermost (27 degrees) of the six concentric rings registered by the mfERG, which corresponds to the area measured by the OCT (psi 3.5 mm). RESULTS: Visual acuity was similar before and after photocoagulation, 1.0; 0.7-1.0 (md, range) versus 1.0; 0.6-1.0 (md, range). The mean values of the ring average amplitudes were reduced in the first and second, third and fourth concentric rings from foveola after photocoagulation, p = 0.001, p = 0.011 and p = 0.004, respectively. No change was seen in implicit time after treatment. OCT values were similar before and after photocoagulation. There was no correlation between retinal thickness assessed with OCT and amplitudes measured by the mfERG. CONCLUSION: In spite of unchanged values of retinal thickness and visual acuity, panretinal photocoagulation seems to cause a functional impairment in the adjacent untreated macula, shown by reduced amplitudes measured by the mfERG. PMID- 15881257 TI - Two components of the human alcohol electro-oculogram. AB - Light onset or drinking alcohol causes the standing potential of the eye to rise and then fall to a trough (the EOG). After allowing for the time for the alcohol to be absorbed into the blood stream, the changes of current with time are identical for the two agents but each acts through a separate pathway, on the same effector mechanism. We have shown that +ve and -ve processes of the alcohol EOG may be differentially affected in disease. We have now determined the separate dose-response relationship of the two voltage changes. Alcohol diluted with water was given by mouth to fasting dark-adapted subjects. Recordings continued until both the positive peak and the later negative trough were well characterised. Doses of alcohol ranged from 3.54 to 450 mg Kg(-1) of body weight. Experiments were carried out on three normal subjects, 4-8th decade. The results are consistent with the hypothesis that each voltage change is determined by the relation: [EtOH] x [R] <--> [EtOH.R], where <--> represents a reversible reaction. For the +ve peak, semi-saturation occurs at approximately 35 mg Kg(-l). For the -ve trough it is smaller, 11 mg Kg(-l). Therefore the result is consistent with there being 2 distinct processes, and the human EOG cannot be a single 'damped oscillation'. During the short period when change of blood alcohol concentration is effective in causing the EOG sequence (using doses which provoke large voltage changes), the computed blood concentration varies from 0.01 to 0.1 mM, i.e. is > 2 orders of magnitude less than the levels required for intoxication. PMID- 15881258 TI - Assessment of macular function by multifocal electroretinogram in diabetic macular edema before and after vitrectomy. AB - The purpose of this study was to assess the effects of vitrectomy on retinal function in macular and paramacular areas in patients with diabetic macular edema (DME). Ten eyes of 9 patients with successful vitrectomy for DME were evaluated by multifocal electroretinogram (mfERG), optical coherence tomography (OCT), and visual acuity preoperatively and 1, 2, 3, 4, 5 and 6 months postoperatively. Compared with pretreatment values, the response of the positive wave (P1) in macular and paramacular areas tended to decrease in latency from the second postoperative month, and increase in amplitude at the third postoperative month. There was no significant change in response of the negative wave (N1). The tendency in the change of macular function is coincident with that of macular morphology. Therefore, The multifocal ERG may provide objective criteria for the functional evaluation of DME before and after vitrectomy. PMID- 15881260 TI - The effect of fixation instability on the multifocal VEP. AB - In multifocal visually evoked potentials (mfVEP), we find reversals in waveform near the horizontal meridian due to convolutions in the cortex. This renders the mfVEP very sensitive to small changes in gaze position. In this study we tested the effects of very small amounts of fixation instability on the mfVEP topography under controlled conditions using four normal subjects. In order to simulate unstable fixation, subjects were instructed to move their fixation point systematically in a clockwise direction between the endpoints of a fixation cross every few seconds (two degree diameter cross = one degree fixation error). Results were compared against a control condition with stable, central fixation. The effects of 0.5 degrees fixation error are small, but 1.0 degrees fixation error can produce a large decrease in root mean square signal amplitude (e.g., 60%) in the central foveal region (i.e., within 1.4 degrees eccentricity). The size of the effect drops off rapidly with eccentricity and varies greatly between areas within a subject, and between the four subjects. Beyond 3.0 degrees eccentricity the effects are minimal. Unstable fixation with relatively small fixation errors caused a dramatic decrease in mfVEP amplitude within three degrees of eccentricity, which can be misinterpreted as loss of macular function. Fixation monitoring is essential to obtain accurate results in the macular area when recording mfVEPs. PMID- 15881259 TI - Pattern electroretinogram for monitoring the efficacy of intravitreal triamcinolone injection in diabetic macular edema. AB - PURPOSE: To investigate the efficacy of intravitreal triamcinolone (IVT) by evaluation of pattern electroretinogram (PERG) in diabetic patients with clinically significant macular edema (CSME). METHODS: Forty eyes of 40 diabetic patients were treated with 8 mg of IVT injection as primary therapy for CSME. The main outcome measures included best-corrected visual acuity, fundus fluorescein angiography, P50 amplitudes of pattern electroretinogram (PERG) and intraocular pressures before and after injection. RESULTS: The mean follow-up time was 6.1 months. Mean visual acuity improved significantly from a mean LogMAR value of 1.14 +/- 0.16 at baseline to a maximum of 0.73 +/- 0.30. The mean baseline P50 amplitude of PERG before intravitreal injection was 1.5 +/- 0.9 microV. After the treatment, it was 2.1 +/- 1.1 microV at 1-month, 2.4 +/- 1.0 microV at 3-month and 2.1 +/- 0.9 microV at last visit and the differences were significant when compared with baseline values (for each, p < 0.001). Temporary increases in intraocular pressures were controlled with topical antiglaucomatous medications, if required. CONCLUSION: IVT injection provides rapid improvement in visual acuity of diabetic patients with CSME that has been supported by P50 amplitudes of PERG. P50 amplitudes of PERG may he used as novel predictive value in the evaluation of the effectiveness of IVT injection. PMID- 15881261 TI - The effect of contrast reduction on pattern-reversal VEPs in suspected multiple sclerosis and optic neuritis. AB - A retrospective study was conducted to investigate whether the use of low contrast stimuli in addition to the standard high-contrast stimuli would increase the sensitivity of the pattern-reversal P100 when testing for multiple sclerosis (MS) and optic neuritis (ON). We found that there was a statistically significant increase in the number of abnormal results when using 25' checks, but not for 50' checks. Examination of patient records showed that VEP abnormalities to low contrast stimuli only was not prognostic for subsequent development of multiple sclerosis. PMID- 15881262 TI - Standardized full-field electroretinography in rabbits. AB - We present a procedure for full-field ERG recording in rabbits, based on the human ERG standards published by the International Society for Clinical Electrophysiology of Vision (ISCEV). Following initial pilot experiments, six animals aged 3 months and 11 animals between 1 and 2 years were investigated. All animals displayed well detectable and reproducible separate cone and rod responses under appropriate stimulus conditions. The b-wave was smaller in young animals than in old, but there were no similar differences in the b-wave implicit times. The animals had to be lightly sedated, which was shown to have no adverse effects on the recordings. Standard deviations of normalized adult rabbit recordings were comparable to human recordings. The measurements were less precise in young animals. We suggest that our procedure is well suited for further scientific studies in this animal model. PMID- 15881263 TI - Motion-onset VEPs to translating, radial, rotating and spiral stimuli. AB - Motion-onset related visual evoked potentials (M-VEPs) were recorded as a result of the three basic (translating, radial and rotating) and one complex (spiral) motion stimulations in five subjects. Low contrast, retinotopically scaled patterns evoked potentials with major motion-onset specific negativity N160 with maximum in the parieto-temporal region. All multidirectional motion stimuli elicited the motion-onset response of significantly higher amplitude and shorter latency compared to the translating (unidirectional) motion. The rotation-onset evoked potentials had significantly shorter latencies than the rest of explored stimuli. The most stable responses with the largest N160 amplitude were recorded to the radial motion. After masking of the central 20 degrees of the visual field these motion-onset VEPs were acquired without statistically significant amplitude drop. The efficiency and usefulness of the radial stimulus is presented in two clinical cases. PMID- 15881264 TI - Determining abnormal interocular latencies of multifocal visual evoked potentials. AB - PURPOSE: To describe methods for measuring interocular latency differences of multifocal visual evoked potentials (mfVEP) and for determining regions with abnormal interocular latencies in patients. METHODS: The mfVEPs from 100 individuals with normal visual fields and normal fundus examinations were analyzed. Individuals ranged in age from 21.6 to 92.4 years. The stimulus was a 60 sector, pattern-reversing dartboard display. Each sector had 16 checks, 8 white (200 cd/m2) and 8 black (< 1 cd/m2). Interocular latency was measured as the temporal shift producing the best cross-correlation value between the corresponding responses of each eye. The 'corrected interocular latency' was defined as the difference between this shift and the mean interocular latency (shift) for a particular sector and recording channel. RESULTS: The variability of the corrected interocular latency decreased as the signal-to-noise ratio (SNR) of the mfVEP responses increased. For example, the 95% confidence intervals decreased from over 16 ms to under 4 ms as SNR increased. Grouping and summing the responses also lead to an increase in SNR and a decrease in the confidence interval. The results of various cluster criteria were also derived. A cluster criterion (e.g. two or more contiguous points within a hemisphere exceeding a given confidence interval), can serve to increase the specificity for detection of eyes or individuals with abnormal interocular latencies. For example, while 21% of the eyes had 3 or more points exceeding the 5% confidence interval, only 1.8% of the eyes had a cluster of 3 or more of these points. Finally, interocular latency was only weakly correlated with age (r = 0.26). CONCLUSION: In testing for abnormalities in interocular latencies, the confidence interval should be based upon the SNR of the response. Grouping and summing responses to increase SNR or employing a cluster test may also prove useful. PMID- 15881265 TI - Determining abnormal latencies of multifocal visual evoked potentials: a monocular analysis. AB - PURPOSE: To describe a methodology for measuring abnormal timing of monocular multifocal visual evoked potentials (mfVEP). METHODS: The mfVEPs from 100 individuals with normal visual fields and normal fundus exams were analyzed. The stimulus was a 60 sector, pattern-reversing dartboard display. For each of the 60 locations of the dartboard and each channel and each eye, a template was derived based upon the average of the responses from the 100 normal individuals. In deriving this template, care was taken to exclude those responses reversed in polarity as compared to the average response. The best array of responses for each individual was compared to these templates. The relative latency of each response was measured as the temporal shift producing the best cross-correlation value. RESULTS: The 95% confidence interval (CI) decreased as the signal-to-noise ratio (SNR) of the mfVEP responses increased. For example, the 95% CI decreased from over 17 ms to under 9 ms as the SNR increased. Grouping and summing the responses also lead to an increase in SNR and a decrease in CI. Because the number of points exceeding the CI is not randomly distributed among normal individuals, a cluster criterion (e.g. two or more contiguous points within a hemisphere exceeding a given confidence interval) can be helpful. For example, while 18% of the eyes had 5 or more points exceeding the 5% confidence interval, only 6.5% of the eyes had a cluster of 5 of these points. The correlation between relative latency and age was relatively low (r = 0.46). CONCLUSION: For detecting abnormalities in the timing of monocular, mfVEP responses, a template method provides a reasonable approach. In devising a particular test for abnormal timing, the CI should be based upon the SNR of the response. In addition, grouping and summing responses to increase SNR or employing a cluster test may also prove useful. PMID- 15881266 TI - Background light adaptation of the retinal neuronal adaptive system. II. Dynamic effects. AB - The dynamic effects of continuous exposure to light on the neuronal adaptive system of the retina, as indicated by the oscillatory response (OPs) of the electroretinogram (ERG) were studied in the albino rat. Digitally filtered OPs and the a- and b-waves of the corneal ERG were simultaneously recorded in dark adaptation, during continuous light adaptation to four levels of background light (BGL) changing in steps of two log units from 1.43 x 10(-6) cd/m2, referred to as 'low and high scotopic, low and high mesopic' levels. Exposed to 'high scotopic' BGL the total oscillatory response (SOP) significantly enhanced within the first minute, whereas the amplitudes of the a- and b-waves were unaffected. In 'low mesopic' BGL the SOP increased within the first minute, whereas the a- and b waves significantly decreased. 'High mesopic' BGL instantaneously and profoundly reduced both the SOP and the slow potentials. The individual OPs changed in amplitudes mainly within the first minute of BGL. In general, the earlier OPs (O1 and O2) reacted more to the two 'scotopic' BGL levels, whereas the later OPs (O3 and 04) were more affected by the relatively brighter two 'mesopic' conditions. In conclusion, the rapid increase of the OPs within the first minute of 'high scotopic' and 'low mesopic' BGL exposure may represent a rudimentary light adaptational effect in the rod-dominated rat retina. These findings also suggest that the neuronal adaptive mechanism of the retina seems to be a robust system, probably attaining preservation of visual abilities in the rat on exposure to light. PMID- 15881267 TI - Sleep paralysis as spiritual experience. AB - This article presents an overview of the sleep paralysis experience from both a cultural and a historical perspective. The robust, complex phenomenological pattern that represents the subjective experience of sleep paralysis is documented and illustrated. Examples are given showing that, for a majority of subjects, sleep paralysis is taken to be a kind of spiritual experience. This is, in part, because of the very common perception of a non-physical 'threatening presence' that is part of the event. Examples from various cultures, including mainstream contemporary America which has no widely known tradition about sleep paralysis, are used to show that the complex pattern and spiritual interpretation are not dependent on cultural models or prior learning. This is dramatically contrary to conventional explanations of apparently 'direct' spiritual experiences, explanations that are summed up as the 'Cultural Source Hypothesis.' This aspect of sleep paralysis was not recognized through most of the twentieth century. The article examines the way that conventional modern views of spiritual experience, combined with medical ideas that labeled 'direct' spiritual experiences as psychopathological, and mainstream religious views of such experiences as heretical if not pathological, suppressed the report and discussion of these experiences in modern society. These views have resulted in confusion in the scientific literature on sleep paralysis with regard to its prevalence and core features. The article also places sleep paralysis in the context of other 'direct' spiritual experiences and offers an 'Experiential Theory' of cross-culturally distributed spiritual experiences. PMID- 15881268 TI - 'The ghost pushes you down': sleep paralysis-type panic attacks in a Khmer refugee population. AB - Among a psychiatric population of Cambodian refugees (N = 100), 42% had current- i.e. at least once in the last year--sleep paralysis (SP). Of those experiencing SP, 91% (38/42) had visual hallucinations of an approaching being, and 100% (42/42) had panic attacks. Among patients with post-traumatic stress disorder (PTSD; n = 45), 67% (30/45) had SP, whereas among those without PTSD, only 22.4% (11/45) had SP (chi2 = 20.4, p < .001). Of the patients with PTSD, 60% (27/45) had monthly episodes of SP. The Cambodian panic response to SP seems to be greatly heightened by elaborate cultural ideas--with SP generating concerns about physical status, 'good luck' status, 'bad luck' status, sorcery assault, and ghost assault--and by trauma associations to the figure seen in SP. Case vignettes illustrate cultural beliefs about, and trauma resonances of, SP. A model to explain the high rate of SP in this population is presented. SP is a core aspect of the Cambodian refugees response to trauma; when assessing Cambodian refugees, and traumatized refugees in general, clinicians should assess for its presence. PMID- 15881269 TI - Cultural variation in the clinical presentation of sleep paralysis. AB - Sleep paralysis is one of the lesser-known and more benign forms of parasomnias. The primary or idiopathic form, also called isolated sleep paralysis, is illustrated by showing how patients from different cultures weave the phenomenology of sleep paralysis into their clinical narratives. Clinical case examples are presented of patients from Guinea Bissau, the Netherlands, Morocco, and Surinam with different types of psychopathology, but all accompanied by sleep paralysis. Depending on the meaning given to and etiological interpretations of the sleep paralysis, which is largely culturally determined, patients react to the event in specific ways. PMID- 15881270 TI - Inuit interpretations of sleep paralysis. AB - Traditional and contemporary Inuit concepts of sleep paralysis were investigated through interviews with elders and young people in Iqaluit, Baffin Island. Sleep paralysis was readily recognized by most respondents and termed uqumangirniq (in the Baffin region) or aqtuqsinniq (Kivalliq region). Traditional interpretations of uqumangirniq referred to a shamanistic cosmology in which the individual's soul was vulnerable during sleep and dreaming. Sleep paralysis could result from attack by shamans or malevolent spirits. Understanding the experience as a manifestation of supernatural power, beyond one's control, served to reinforce the experiential reality and presence of the spirit world. For contemporary youth, sleep paralysis was interpreted in terms of multiple frameworks that incorporated personal, medical, mystical, traditional/shamanistic, and Christian views, reflecting the dynamic social changes taking place in this region. PMID- 15881271 TI - Sleep paralysis, sexual abuse, and space alien abduction. AB - Sleep paralysis accompanied by hypnopompic ('upon awakening') hallucinations is an often-frightening manifestation of discordance between the cognitive/perceptual and motor aspects of rapid eye movement (REM) sleep. Awakening sleepers become aware of an inability to move, and sometimes experience intrusion of dream mentation into waking consciousness (e.g. seeing intruders in the bedroom). In this article, we summarize two studies. In the first study, we assessed 10 individuals who reported abduction by space aliens and whose claims were linked to apparent episodes of sleep paralysis during which hypnopompic hallucinations were interpreted as alien beings. In the second study, adults reporting repressed, recovered, or continuous memories of childhood sexual abuse more often reported sleep paralysis than did a control group. Among the 31 reporting sleep paralysis, only one person linked it to abuse memories. This person was among the six recovered memory participants who reported sleep paralysis (i.e. 17% rate of interpreting it as abuse-related). People rely on personally plausible cultural narratives to interpret these otherwise baffling sleep paralysis episodes. PMID- 15881272 TI - Sleep paralysis in African Americans with panic disorder. AB - Studies have reported a wide range in lifetime prevalence of sleep paralysis (SP). This variation may stem from cultural factors, stressful life events and genetic differences in studied populations. We found that recurrent SP was more common among African-American participants, especially those with panic disorder. Recurrent SP was reported by 59% of African Americans with panic disorder, 7% of whites with panic disorder, 23% of African-American community volunteers and 6% of white community volunteers. Significantly more early life stressors were reported by African Americans than whites. Higher levels of psychosocial stressors, including poverty, racism and acculturation, may contribute to the higher rates of SP experienced by African Americans. PMID- 15881273 TI - Prevalence and illness beliefs of sleep paralysis among Chinese psychiatric patients in China and the United States. AB - To investigate the prevalence and illness beliefs of sleep paralysis (SP) among Chinese patients in a psychiatric out-patient clinic, consecutive Chinese/Chinese American patients who attended psychiatric out-patient clinics in Boston and Shanghai were asked about their lifetime prevalence, personal experience and perceptions regarding the causes, precipitating factors, consequences, and help seeking of SP. During the 4-month study period, 42 non-psychotic psychiatric out patients from the Boston site and 150 patients from the Shanghai site were interviewed. The prevalence of SP was found to be 26.2% in Boston and 23.3% in Shanghai. Patients with post-traumatic stress disorder (PTSD) or panic disorder reported a higher prevalence of SP than did patients without these disorders. Patients attributed SP to fatigue, stress, and other psychosocial factors. Although the experience has traditionally been labeled 'ghost oppression' among the Chinese, only two patients, one from each site, endorsed supernatural causes of their SP. Sleep paralysis is common among Chinese psychiatric out-patients. The endorsement of supernatural explanations for SP is rare among contemporary Chinese patients. PMID- 15881274 TI - Interstitial lung diseases--where we started from and are now going. AB - BACKGROUND AND AIM: Research into mechanisms causing interstitial lung diseases (ILD) began 35 years ago with the advent of cellular immunology and techniques to sample airways for biologic materials. After an analysis of lung research programs by the then National Heart and Lung Institute in 1972 identified as a priority the study of fibrotic and immunologic lung diseases, this began in the Pulmonary Branch (1974) of the Institute's intramural program. The Division of Lung Diseases initiated extramural research support also. ILD research developed quickly at many centers in the US and throughout the world. This review focuses on idiopathic pulmonary fibrosis (IPF) and highlights some of the initial research from the Pulmonary Branch. RECENT RESEARCH PARADIGM: In the 1990s research emphasis changed from a focus on inflammation to alveolar epithelial injury, fibrogenesis in fibroblastic foci, myofibroblast function, cytokine secretion and disordered matrix remodeling. More precise classification of ILD was advocated, especially for IPF. New strategies for therapy of IPF followed, including anti-fibrotic agents and interferon gamma treatment. However, therapy is still not sufficiently effective. Much is still left to do. FUTURE DIRECTIONS: The NHLBI research support continues for ILD, especially IPF. Current programs include: searching for new molecular therapeutic targets; establishing of a clinical network for IPF patients to assess combinations of therapy and new agents as appropriate; identifying genomic and genetic susceptibility factors; and creating a repository for lung tissue and biologic samples to aid investigators. PMID- 15881275 TI - CD4+CD28-T cells are expanded in sarcoidosis. AB - BACKGROUND AND AIM: A subset of CD4+ lymphocytes lacking CD28, an important costimulatory molecule, is increased in certain inflammatory conditions. However, studies have not directly studied CD4+CD28-lymphocytes in patients with chronic sarcoidosis. The aim of this study was to further characterize the CD4+CD28-T cell population in patients with sarcoidosis, particularly those with active disease. METHODS: Seventeen patients with chronic sarcoidosis and 15 blood donors were studied. Bronchoalveolar lavage cells were available for paired analysis in seven sarcoid patients. In 4 sarcoid patients, adequate sample was available for intracellular cytokine analysis by flow cytometry. IFN-gamma production in plasma and BAL was determined by ELISA and cytometric bead array analysis and compared to previously studied controls. RESULTS: Peripheral blood from patients with sarcoidosis had a significantly higher proportion of CD4+CD28- cells compared with healthy donors. A higher percentage of CD4+CD28- cells was evident in the BAL relative to peripheral blood in patients with active sarcoid. IFN-gamma levels were greater both in the plasma and concentrated BAL fluid of sarcoid subjects compared to controls. The majority of IFN-gamma and TNF-alpha producing lymphocytes were CD28+ in both healthy blood donors and sarcoid subjects. CONCLUSIONS: CD4+CD28- cells are increased in the peripheral blood and lungs of patients with sarcoidosis requiring treatment. These cells may contribute to the inflammatory response; however, they are not major contributors of IFN-gamma or TNF-alpha. PMID- 15881276 TI - Variations in alveolar cell populations, lymphocyte subsets and NK-cells in different stages of active pulmonary sarcoidosis. AB - BACKGROUND: The type and mode of presentation of pulmonary sarcoidosis (radiologic stage, activity) influence alveolar lymphocyte number and subsets as well as other inflammatory cells. AIM: To investigate the variations in alveolar cells, lymphocyte subsets and NK-cells in different radiological stages of active pulmonary sarcoidosis. METHODS: 85 untreated, newly diagnosed patients (32 male, 53 female), median age 47.2+/-14.7 years were investigated. Patients were classified into chest x-ray stages (33 stage I, 27 stage II, 14 stage III and 11 stage IV disease). Bronchoalveolar lavage was performed with 4 portions of 50 mL. Total cells and cell differentials were counted, while CD3+, CD4+, CD8+, CD19+, CD3-CD16/56+ (NK-cells) and CD3+CD16/56+ (NKT-cells) were determined by flow cytometry. RESULTS: Lymphocytosis was high in all stages. Significant differences were found in: a) CD4 between stages I and II and I and IV. b) CD8 between stages I and IV and c) NK and NKT cells between stages I and III. CONCLUSIONS: Variations were observed in alveolar cells and lymphocyte subsets in different stages of pulmonary sarcoidosis. Cells exhibiting cytotoxic activity were increased with stage progression, CD8 from stage I to IV, NK and NKT cells from stage I to III. These cells may be involved in the inflammatory process regulating granuloma formation. PMID- 15881277 TI - Cytotoxic T-lymphocyte antigen 4 gene polymorphisms in sarcoidosis patients. AB - BACKGROUND AND AIM: Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is a co-stimulatory molecule that is expressed on activated T-cells, and is an important regulator of T-cell activation in T-cell-dependent immune responses. CTLA-4 signals lead to the downregulation of T-cell proliferation and activation. Sarcoidosis is a systemic granulomatous disorder of unknown etiology, which shows abnormal regulation of T cell activation. Polymorphisms of CTLA-4 have been reported to alter CTLA-4 expression, and are associated with many diseases. In the present study we investigated CTLA-4 polymorphisms of promoter -318(C/T) and exon 1 +49(A/G) in sarcoidosis patients and control subjects to determine whether these genetic variations affect sarcoidosis. METHODS: One hundred and six sarcoidosis patients and 100 healthy control subjects were studied. The polymerase chain reaction technique and direct genomic sequencing were used to determine the genotypes of promoter -318(C/T) and exon 1 +49(A/G) in the CTLA-4 gene. RESULTS: We found no difference in the distribution of either genotype between the healthy control subjects and sarcoidosis patients. Among the sarcoidosis patients, the 318(C/T) CC genotype (p = 0.011) and AG or GG genotype at position +49(A/G) (p = 0.004) were increased with ocular involvement compared with those patients without ocular involvement. Furthermore, the +49(A/G) GG genotype was increased in patients with three or more organs affected compared with patients with fewer organs affected (p = 0.019). CONCLUSIONS: The CTLA-4 polymorphisms are not associated with disease susceptibility of sarcoidosis, but these genetic variations significantly influence phenotypes of sarcoidosis. PMID- 15881278 TI - Propionibacterium acnes is the most common bacterium commensal in peripheral lung tissue and mediastinal lymph nodes from subjects without sarcoidosis. AB - BACKGROUND: In studies of the unknown etiology of sarcoidosis, Propionibacterium acnes (a possible agent) was found in the lungs and lymph nodes of many sarcoidosis patients and some control subjects. P. acnes might be commensal not only to the skin, conjunctivae, and intestine, but also to the lungs and lymph nodes of individuals without sarcoidosis. METHODS: We cultured peripheral lung tissue and various lymph nodes obtained from patients with diseases other than sarcoidosis. DNA of 45 isolates of P. acnes from these patients, 67 isolates from normal skin, conjunctiva, and intestine, and 39 isolates from sarcoid lymph nodes were compared by random amplified polymorphic DNA analysis. RESULTS: P. acnes was isolated from half of 43 lungs and 8 of 11 mediastinal lymph nodes, mostly in pure culture. P. acnes was isolated from half of 20 gastric and 3 of 12 intestinal lymph nodes; intestinal bacteria were also numerous. In general, fewer than 500 colony-forming units of P. acnes per gram tissue were isolated, but 4 lung tissue specimens, 2 of which had a few granulomas, had many more. P. acnes strains from a particular site (lung, lymph node, skin or conjunctivae, and intestine) were genetically similar, more than isolates obtained from different sites. Lymph-node isolates from subjects with and without sarcoidosis differed little. CONCLUSION: These results suggest that P. acnes normally resides in peripheral lung tissue and mediastinal lymph nodes and that the strains of P. acnes isolated from sarcoid lymph nodes were not specific to sarcoidosis. PMID- 15881279 TI - Association of small fiber neuropathy with cardiac sympathetic dysfunction in sarcoidosis. AB - BACKGROUND AND AIM: Recently we found that small fiber neuropathy (SFN) occurs frequently in sarcoidosis. Autonomic dysfunction may be a feature of SFN. Since cardiac autonomic dysfunction has been identified as a strong predictor of morbidity and mortality, recognition of cardiac autonomic involvement is of clinical relevance. It was hypothesised that SFN might be related to cardiac sympathetic denervation in sarcoidosis. METHODS: In the present study 45 consecutive sarcoidosis patients (13 without SFN assessed by thermal threshold testing (TTT), 32 with SFN (abnormal TTT) were enrolled. To rule out bias due to myocardial ischemia, cases with abnormal Thallium (201Tl) perfusion scintigraphy were excluded (n = 2). Cardiovascular autonomic function testing (Ewing tests) and 123I-MIBG (metaiodobenzylguanidine) scintigraphy were used to assess cardiac autonomic function. Further cardiac diagnostic work-up included ECG, Holter recording and echo Doppler cardiography. RESULTS: Mild to moderate heterogeneity of 123I-MIBG uptake regional in the myocardium was demonstrated in a substantial number of the studied sarcoidosis population, especially in those with SFN (abnormal TTT). Mean inferior-anterior ratios were 0.85+/-0.17 (SFN) and 1.0+/ 0.17 (no SFN; p = 0.003), respectively. Four out of the 14 cases with abnormal MIBG scintigraphy and SFN showed an abnormal Ewing test. CONCLUSION: Cardiac sympathetic dysfunction assessed by use of 123I-MIBG myocardial scanning appeared to be heterogeneous in sarcoidosis patients and dependent on the presence or absence of SFN. MIBG scintigraphy may be of additional value in the management and follow-up of sarcoidosis patients. Future study is warranted to explore possible prognostic and therapeutic implications of these findings in sarcoidosis. PMID- 15881280 TI - Analysis of KL-6 and SP-D as disease markers in bird fancier's lung. AB - BACKGROUND AND AIM: KL-6 and SP-D are potential serum markers in interstitial lung diseases. Their discriminative value, and ability to reflect pulmonary disease activity and prognosis in bird fancier's lung were analyzed. METHODS: We studied 49 patients, 38 unexposed and 9 exposed controls. Serum KL-6 and SP-D concentrations were measured at presentation and a second sample, taken after antigen avoidance, was available in 17 patients. Pulmonary function tests were analyzed at presentation and 2-year follow-up. RESULTS: KL-6 and SP-D were significantly elevated in patients compared to controls (p < 0.0001). ROC curve analysis revealed that both are equally useful in discriminating patients from controls. Analysis of their value as activity markers showed that both correlated with pulmonary function impairment; however, KL-6 correlated best with diffusing capacity. Evaluation of their predictive value showed that higher levels at onset were associated with improvement of diffusing capacity during follow-up. Further, it was noted that KL-6 and SP-D levels decreased after more than one month of allergen avoidance. CONCLUSIONS: KL-6 and SP-D appear useful serum markers in bird fancier's lung. Since higher levels are associated with more severe lung function impairment at presentation, and better recovery over time, we postulate that in this disease they are especially markers of disease activity. PMID- 15881281 TI - Pleural involvement in chronic sarcoidosis detected by thoracic CT scanning. AB - BACKGROUND AND AIM: 5-10% of patients with sarcoidosis exhibit pleural involvement by standard chest radiograph (CXR) usually associated with chronic advanced lung disease. The frequency of pleural disease in sarcoidosis by chest CT scan is unknown. This study compared pleural involvement by standard CXR with thoracic CT scan and assessed the impact of pleural involvement on pulmonary function tests (PFT) in patients. METHODS: The records of 61 consecutive patients seen in the Sarcoidosis Service at Mount Sinai Hospital who had thoracic CT scan, standard CXR, and recent PFT were reviewed. RESULTS: 25 of the 61 patients (41%) had pleural involvement by CT (20 thickening, 5 effusions), compared to 7 (11%) by standard CXR (3 thickening, 4 effusions). Bilateral pleural thickening was more commonly seen in patients with CT evidence of parenchymal fibrosis. On univariate analysis, CT evidence of parenchymal fibrosis and CT pleural thickening were significantly associated with an increased odds of restrictive PFTs, ORs of 7.49 (CI 1.7-31.8) and 4.1 (CI 1.32-12.7), respectively. The association between CT pleural thickening and restrictive PFTs lost significance when adjusted for the confounding effect of parenchymal fibrosis. Restrictive physiology was associated with CT evidence of parenchymal fibrosis even when adjusted for pleural thickening (OR = 5.35 CI = 1.18-24.2). CONCLUSION: Sarcoidal pleural involvement as detected by CT scan is much more common than by CXR and is associated with restrictive pulmonary dysfunction. Pleural thickening was also associated with CT evidence of pulmonary fibrosis but not restrictive physiology when adjusted for parenchymal scarring. PMID- 15881282 TI - Relationship between basal thinning of the interventricular septum and atrioventricular block in patients with cardiac sarcoidosis. AB - BACKGROUND: Basal thinning of the interventricular septum (IVS) and atrioventricular block (AVB) are characteristic features of cardiac sarcoidosis. Since the conduction system passes along IVS, it has been considered that a close connection exists between basal thinning of IVS and AVB. However, neither the incidence of cases showing basal thinning of IVS nor the relation between it and AVB has been clarified. We thus investigated to elucidate these two issues. METHODS: Thirty-five patients with cardiac sarcoidosis were selected for this study and underwent echocardiographic examination. The wall thickness of IVS was measured at a site 1 cm below the aortic valve inserted point of IVS. Thickness of this site < or = 5 mm was defined as thinning. Twelve-lead and Holter electrocardiograms were obtained to determine the presence/absence and degree of AVB. RESULTS: Basal thinning of IVS was noted in 7 of the 35 patients (20%). AVB was present in 4 of these 7 (57%), and was first degree in 3 (43%) and third degree in one (14%). AVB was not present in 3 patients. Basal thinning of IVS was not apparent in 28 of the 35 patients (80%). AVB was observed in 14 of the 28 patients, 3 had first degree block, 2 had second degree block, and 9 had third degree block. AVB was not observed in 14 of the 28 patients. CONCLUSIONS: These results clarified that basal thinning of IVS is not as common as previously thought in cardiac sarcoidosis, basal thinning of IVS and the presence/absence and degree of AVB are not necessarily correlated. PMID- 15881283 TI - Clinical course of sarcoidosis in dependence on HLA-DRB1 allele frequencies, inflammatory markers, and the presence of M. tuberculosis DNA fragments. AB - BACKGROUND AND AIM: For sarcoidosis it is generally hypothesized that inherited factors and environmental antigens may contribute to pathogenesis. Since M. tuberculosis DNA was found in a significant percentage of sarcoidosis patients, we analyzed the relationship between HLA-DRB1 alleles, inflammatory markers and the presence of M. tuberculosis DNA in sarcoidosis and its influence on clinical course. METHODS: From 144 patients with sarcoidosis lung tissue, BAL and/or blood were investigated by means of PCR assays to detect an 123 bp multicopy element of M. tuberculosis DNA and HLA-DRB1 alleles, respectively. ACE was measured spectrophotometrically, sIL-2R by ELISA. Clinical data describing the disease course were available from 63 patients. RESULTS: The percentage of M. tuberculosis positive sarcoidosis patients was significantly increased in the chronic patients group compared to acute disease. The percentage of HLA-DRB 1*03 positive patients was significantly higher in acute sarcoidosis, whereas in chronic disease the HLA-DRB1*11 positive patients were clearly over-represented. In addition, we found a highly significant correlation of HLA-DRB1*11 or -DRB1*15 alleles and/or the presence of M. tuberculosis DNA to a chronic disease course, whereas HLA-DRB1*03 or -DRB1*04 alleles combined with the absence of M. tuberculosis DNA were associated with an acute sarcoidosis (p = 0.009). ACE and sIL2-R serum levels were significantly higher in M. tuberculosis positive sarcoidosis independent of the HLA-DRB1 specificity, but did not differ between acute and chronic disease course alone. CONCLUSIONS: The association between certain HLA-DR antigens, the presence of M. tuberculosis DNA and disease course indicates that specific antigens of M. tuberculosis may play a pathogenetic role in chronic sarcoidosis. PMID- 15881284 TI - Bilateral Gasser's ganglion sarcoidosis: diagnosis, treatment and unsolved questions. AB - Magnetic resonance imaging is currently the best means for confirming clinical suspicion of neurosarcoidosis as well as being useful in the follow-up of corticosteroid-treated patients. We report the case of a 34 year old male presenting suspected Heerfordt's syndrome with concentric facial hypesthesia. Mediastinal and parotid sarcoidosis was diagnosed and magnetic resonance imaging showed bilateral sarcoid involvement of Gasser's ganglion cisternae (such involvement was not revealed by computed tomography). The patient received corticosteroid therapy, with a clinical and radiological improvement. Magnetic resonance imaging showed disappearance of Gasser's ganglion lesions despite the persistence of mild facial hypesthesia. This case is noteworthy for its extremely rare lesion site. Post-treatment discrepancy between the clinical picture and imaging results is probably due to low MRI resolution threshold. 18-FDG positron emission tomography imaging might perhaps overcome the limits of magnetic resonance imaging. PMID- 15881285 TI - Sildenafil in pulmonary hypertension. PMID- 15881286 TI - Sarcoidosis presenting as isolated intraparotid lymphadenopathy. PMID- 15881287 TI - Cognitive gerontology: cognitive change in old age. Introduction. PMID- 15881288 TI - Biomarkers, health, lifestyle, and demographic variables as correlates of reaction time performance in early, middle, and late adulthood. AB - We aimed to identify demographic, health, and biomarker correlates of reaction time performance and to determine whether biomarkers explained age differences in reaction time performance. TIhe sample comprised three representative cohorts aged 20-24, 40-44, and 60-64 years, including a total of 7,485 participants. Reaction time measures of intraindividual variability and latency were used. The measure of intraindividual variability used was independent of mean reaction time. Older adults were more variable than younger adults in choice reaction time performance but not simple reaction time performance. The most important correlates of reaction time performance after gender and education were biological markers such as forced expiratory volume at one second, grip strength, and vision. Few measures of physical or mental health or lifestyle were associated with poorer performance on reaction time measures. Biomarkers explained the majority of age-related variance in simple reaction time and a large proportion of variance in choice reaction time. We conclude that for the ages studied, biomarkers are more important than health factors for explaining age differences in reaction time performance. PMID- 15881289 TI - Hearing loss and perceptual effort: downstream effects on older adults' memory for speech. AB - A group of older adults with good hearing and a group with mild-to-moderate hearing loss were tested for recall of the final three words heard in a running memory task. Near perfect recall of the final words of the three-word sets by both good- and poor-hearing participants allowed the inference that all three words had been correctly identified. Nevertheless, the poor-hearing group recalled significantly fewer of the nonfinal words than did the better hearing group. This was true even though both groups were matched for age, education, and verbal ability. Results were taken as support for an effortfulness hypothesis: the notion that the extra effort that a hearing-impaired listener must expend to achieve perceptual success comes at the cost of processing resources that might otherwise be available for encoding the speech content in memory. PMID- 15881290 TI - Age-related declines in context maintenance and semantic short-term memory. AB - This study reports age-related declines in context maintenance (Braver et al., 2001) and semantic short-term memory (STM) and evidencc for a relation between the two. A group of younger and older adults completed a context maintenance task (AX-CPT), a semantically oriented STM task (conceptual span), a phonologically oriented STM task (digit span), and a meaning integration task (semantic anomaly judgement). In the AX-CPT task, a target response is required to the probe letter "X" but only when it is preceded by the letter "A" (the context). Either three (short interference) or six distractor letters (long interference) were presented between the cue and the probe. Results indicated an age-related deficit in context maintenance. Age-related declines were also observed for conceptual span and semantic anomaly judgement but not for digit span. Context maintenance was correlated with conceptual span and semantic anomaly judgement but not with digit span. These correlations were largely mediated by age differences, which also explained variance that was unique to (and not shared among) context maintenance, conceptual span, and semantic anomaly judgement. PMID- 15881291 TI - Memory for proper names in old age: a disproportionate impairment? AB - A common complaint of older adults is that they have trouble remembering names, even the names of people they know well. Two experiments examining this problem are reported in the present article. Experiment I tested episodic memory for surnames and occupations; older adults and younger adults under divided attention performed less well than did full attention younger adults, but showed no disproportionate loss of name information. Experiment 2 examined the ability to name photographs of public figures and of uncommon objects; this experiment therefore tested retrieval from semantic memory. In this case adults in their 70s did show an impairment in recall of names of known people, but not of known objects. Further analyses revealed systematic relations between naming, recognition, and rated familiarity of the categories used. Familiarity largely determined the proportions of recognizable items that were named in a prior phase. Overall, little evidence was found for a disproportionate age-related impairment in naming in either episodic or semantic memory. PMID- 15881292 TI - Searching from the top down: ageing and attentional guidance during singleton detection. AB - Previous investigations of adult age differences in visual search suggest that an age-related decline may exist in attentional processes dependent on the observer's knowledge of task-relevant features (top-down processing). The present experiments were conducted to examine age-related changes in top-down attentional guidance during a highly efficient form of search, singleton detection. In Experiment 1 reaction times to detect targets were lower when target features were constant (feature condition) than when target features were allowed to vary between trials (mixed condition), and this reaction time benefit was similar for younger and older adults. Experiments 2 and 3 investigated possible interactions between top-down and bottom-up (stimulus-driven) processes. Experiment 2 demonstrated that search times for both age groups could be improved when targets varied on an additional feature from distractors (double-feature condition) but only when top-down control was available (feature search). In Experiment 3, the availability of top-down guidance enabled both younger and older adults to override the distracting effects of a noninformative spatial location cue. 'l'hese findings indicate that top-down attentional control mechanisms interact with bottom-up processes to guide search for targets, and that in the context of singleton detection these mechanisms of top-down control are preserved for older adults. PMID- 15881293 TI - Age-related deficits in free recall: the role of rehearsal. AB - Age-related deficits have been consistently observed in free recall. Recent accounts of episodic memory suggest that these deficits could result from differential patterns of rehearsal. In the present study, 20 young and 20 older adults (mean ages 21 and 72 years, respectively) were presented with lists of 20 words for immediate free recall using the overt rehearsal methodology. The young outperformed the older adults at all serial positions. There were significant age related differences in the patterns of overt rehearsals: Young adults rehearsed a greater number of different words than did older adults, they rehearsed words to more recent serial positions, and their rehearsals were more widely distributed throughout the list. Consistent with a recency-based account of episodic memory, age deficits in free recall are largely attributable to age differences in the recency, frequency, and distribution of rehearsals. PMID- 15881294 TI - The attraction effect in decision making: superior performance by older adults. AB - Previous work showed that older adults' choice performance can be wiser than that of younger adults (Tentori, Osherson, Hasher, & May, 2001). We contrasted two possible interpretations: a general expertise/wisdom view that suggests that older adults are generally more skilled at making decisions than younger adults and a domain-specific expertise view that suggests that older adults are more skilled decision makers only in domains in which they have greater knowledge. These hypotheses were contrasted using attraction effect tasks in two different domains: earning extra credit in a course and grocery shopping, domains presumed to be of different levels of knowledge to younger and older adults. Older adults showed consistent choice for both domains; younger adults showed consistent choice only for the extra credit problem. Several explanations of these findings are considered, including Damasio's somatic marker theory and age differences in reliance on heuristic versus analytic styles. PMID- 15881295 TI - Ageing and switching of the focus of attention in working memory: results from a modified N-back task. AB - We conducted two experiments using a modified version of the N-Back task. For younger adults, there was an abrupt increase in reaction time of about 250 ms in passing from N = 1 to N > 1, indicating a cost associated with switching of the focus of attention within working memory. Response time costs remained constant over the range N = 2 to N = 5. Accuracy declined steadily over the full range of N (Experiment 1). Focus switch costs did not interact with either working memory updating (Experiment 1), or global task switching (Experiment 2). There were no age differences in RT costs once general slowing was taken into account, but there was a larger focus-switch-related accuracy cost in older adults than in younger adults. No age sensitivity was found for either updating or global task switching. The results suggest (a) that focus switching is a cognitive primitive, distinct from task switching and updating, and (b) that focus switching shows a specific age-related deficit in the accuracy domain. PMID- 15881296 TI - Processing speed, executive function, and age differences in remembering and knowing. AB - A group of young (n = 52, M = 23.27 years) and old (n = 52, M = 68.62 years) adults studied two lists of semantically unrelated nouns. For one list a time of 2 s was allowed for encoding, and for the other, 5 s. A recognition test followed where participants classified their responses according to Gardiner's (1988) remember-know procedure. Age differences for remembering and knowing were minimal in the faster 2-s encoding condition. However, in the longer 5-s encoding condition, younger persons produced significantly more remember responses, and older adults a greater number of know responses. This dissociation suggests that in the longer encoding condition, younger adults utilized a greater level of elaborative rehearsal governed by executive processes, whereas older persons employed maintenance rehearsal involving short-term memory. Statistical control procedures, however, found that independent measures of processing speed accounted for age differences in remembering and knowing and that independent measures of executive control had little influence. The findings are discussed in the light of contrasting theoretical accounts of recollective experience in old age. PMID- 15881297 TI - Contributions of processing ability and knowledge to verbal memory tasks across the adult life-span. AB - This study investigated the relationships of processing capacity and knowledge to memory measures that varied in retrieval difficulty and reliance on verbal knowledge in an adult life-span sample (N = 341). It was hypothesized that processing ability (speed and working memory) would have the strongest relationship to tasks requiring active retrieval and that knowledge (vocabulary ability) would be related to verbal fluency and cued recall, as participants relied upon verbal knowledge to retrieve category items (fluency) or develop associations (cued recall). Measurement and structural equation models were developed for the entire sample and separately for younger (aged 20-54 years, n = 168) and older (aged 55-92 years, n = 173) subgroups. In accordance with the hypotheses, processing ability was found to be most highly related to free recall, with additional significant relationships to cued recall, verbal fluency, and recognition. Knowledge was found to be significantly related only to verbal fluency and to cued recall. Moreover, knowledge was more important for older than for younger adults in mediating variance in cued recall, suggesting that older adults may use age-related increases in knowledge to partially compensate for processing declines when environmental support is available in memory tasks. PMID- 15881298 TI - Dentistry in crisis. PMID- 15881299 TI - Why be down in the mouth? Three decades of research in oral microbiology. AB - This paper describes some of the work done in the author's laboratory over the past 35 years. The research covers the following areas: the physiology of oral streptococci and their interactions; the physiology of some Gram-negative anaerobes and their interactions in relation to periodontal diseases; preventing the major dental diseases; and the future of oral microbiology. PMID- 15881300 TI - Eating disorders and oral health: a review of the literature. AB - This article is a review of the recent literature pertaining to the oral sequelae of eating disorders (EDs). Dentists are recognized as being some of the first health care professionals to whom a previously undiagnosed eating disorder patient (EDP) may present. However, despite the prevalence (up to 4 per cent) of such conditions in teenage girls and young adult females, there is relatively little published in the recent literature regarding the oral sequelae of EDs. This compares unfavourably with the attention given recently in the dental literature to conditions such as diabetes mellitus, which have a similar prevalence in the adult population. The incidence of EDs is increasing and it would be expected that dentists who treat patients in the affected age groups would encounter more individuals exhibiting EDs. Most of the reports in the literature concentrate on the obvious clinical features of dental destruction (perimolysis), parotid swelling and biochemical abnormalities particularly related to salivary and pancreatic amylase. However, there is no consistency in explanation of the oral phenomena and epiphenomena seen in EDs. Many EDPs are nutritionally challenged; there is a relative lack of information pertaining to non-dental, oral lesions associated with nutritional deficiencies. PMID- 15881301 TI - Dental caries experience in Australian Army recruits 2002-2003. AB - BACKGROUND: Recent data have suggested that the trend of decreasing caries experience in Australian children is slowing with increasing dmft and DMFT scores seen in children. However, there are limited data on dental caries experience in young Australian adults. METHODS: A cross-sectional study of 973 Australian Army recruits was conducted between November 2002 and March 2003. A clinical examination with bitewing radiographs was conducted and a questionnaire was used to elicit socio-demographic information. RESULTS: Mean DMFT scores were 2.43, 3.44, 5.48, 7.02 and 10.77 for subjects aged 17-20, 21-25, 26-30, 31-35 and 36-51 years respectively. Subjects with a lifetime exposure to fluoridated drinking water had a mean DMFT of 2.80 while subjects with no exposure to fluoridated drinking water had a mean DMFT of 3.91. Multivariate Poisson regression found that age, level of educationand lifetime exposure to fluoridated drinking water had a statistically significant effect on caries experience. CONCLUSIONS: It appears that there has been a continual decline in caries experience and prevalence in young Australian adults between 1996 and 2002-2003. Lifetime exposure to fluoridated drinking water conferred an appreciable benefit for subjects in this study compared with subjects with no exposure to fluoridated drinking water. PMID- 15881302 TI - Chronic orofacial pain is associated with psychological morbidity and negative personality changes: a comparison to the general population. AB - BACKGROUND: Chronic orofacial pain is a biopsychosocial problem. Pain description and intensity have been previously reported by the authors. This follow up study reports on the presence and severity of psychological morbidity presence and alseverity changes associated with chronic and personality changes associated with chronic orofacial pain. METHODS: A total of 415 questionnaires for psychological morbidity (238 chronic orofacial pain patients and 175 controls) and 205 responses for personality changes (105 pain patients and 100 controls) were analyzed. Demographic and status, level of education include and current work status. status, level of education and current work status. Psychological variables tested were depression, anger, fear, distress, frustration and anxiety. Pain patients indicated descriptors of their personalities 'pre-pain' and 'with pain'. RESULTS: The chronic pain group reported higher levels of 'feeling sad or miserable' p < 0.001 'feeling frustrated' p = 0.001 and 'feeling anxious, worried' p = 0.022 than the control group. Within the chronic pain group, patients unemployed due to pain or other reasons reported higher levels of 'feeling sad or miserable' and 'feeling frustrated' (p < 0.05) compared with patients engaged in full or part-time work. Negative personality changes due to pain were clearly evident with 'irritable' and 'sad' being frequently chosen words (p < 0.001). CONCLUSIONS: Patients with chronic orofacial pain suffer from negative psychological and personality changes. PMID- 15881303 TI - Oral cancer at a Tasmanian tertiary referral clinic, the Royal Hobart Hospital 1996-2002. AB - BACKGROUND: The Holman Clinic at the Royal Hobart Hospital includes a multi disciplinary head and neck clinic which functions as a tertiary referral centre for Southern Tasmania and involves Ear Nose and Throat surgeons, Oral and Maxillofacial Surgeons, Plastic and Reconstructive Surgeons, Radiation Oncologists and Medical Oncologists. METHODS: The aim of this study was to examine retrospectively the number, gender distribution, age, site of lesion, histology, mortality and treatment modalities of the oral cancers referred to the Holman clinic at the Royal Hobart Hospital. The medical histories and a database of the Holman clinic were used as the sources of data for this study. A total of 101 patients were treated for oral cancer in the Holman clinic at the Royal Hobart Hospital from 1996 to 2002. There were 64 males and 37 females. RESULTS: The distribution of anatomical sites of the oral cancers in this study was as follows: 36 oral tongue lesions, 17 floor of mouth, 13 lip, five retromolar trigone, five mandibular alveolus, six buccal mucosa, nine palatal and 10 minor and major salivary gland cancers. The most common site of oral cancer was the tongue (35.6 per cent), followed by the floor of mouth (16.8 per cent) and lip (12.9 per cent). CONCLUSIONS: The majority of oral cancers were squamous cell carcinoma, except for the salivary gland cancers. The incidence of squamous cell carcinoma was between 67 and 100 per cent, depending upon the site involved. The trends found in this study are similar to those previously documented over the past 20 years. PMID- 15881304 TI - Factors influencing the diagnosis and treatment of periodontal disease by dental practitioners in Victoria. AB - BACKGROUND: Healthy periodontal tissues are essential to overall dental health. Therefore, the detection and management of periodontal disease is an integral part of general dental practice. The aim of this study was to investigate confidence in diagnosis and management of periodontal disease by general dental practitioners (GDPs), assess if the Dental Practice Board guidelines on periodontal record keeping are being addressed, and, if necessary, try to find ways of improving the periodontal knowledge of GDPs. METHODS: A survey assessing practitioner confidence in diagnosing and treating periodontal disease was sent to a random selection of 550 dental care providers registered with the Dental Practice Board of Victoria. RESULTS: Two hundred and eighty five (51.8 per cent) of questionnaires were returned completed. It was found that 79.7 per cent of the sampled population screened all new patients for periodontal disease. The majority of respondents felt confident to diagnose and treat gingivitis and initial periodontitis. However, only 61.9 per cent felt confident to diagnose aggressive/early onset periodontitis, and many were not confident in treating advanced periodontitis (36.3 per cent) or aggressive periodontitis (51.6 per cent). The majority of dentists reported that they provided most of the non surgical periodontal therapy to their patients, while most surgical treatments were referred to specialist periodontists. Factors deemed to be important in influencing the decision to provide periodontal treatment included level of training and ability to motivate patients to improve oral hygiene. Many responents requested periodontic continuing education (CE) courses be run. CONCLUSIONS: Most of the dentists surveyed were confident to diagnose periodontal disease and to treat the more common presentations of periodontal disease. There is some evidence to suggest that some practitioners are not following the minimum requirements set by the Dental Practice Board of Victoria in relation to periodontal record keeping. The results also indicate a need for more periodontic CE courses in Victoria. PMID- 15881305 TI - Three-dimensional modelling and finite element analysis of the human mandible during clenching. AB - BACKGROUND: Until recently, very few papers have been published concerning the development, analysis and experimental verification of three-dimensional, finite element modelling of the human adult edentulous mandible. The purpose of this study was to improve the method of modelling by using computer-aided engineering (CAE) and computer-aided design (CAD) methods and to utilize the model in analyzing maxillofacial problems. METHODS: The model geometry was derived from position measurements taken from 28 diamond blade cut cross-sections of an average size human adult edentulous mandible and generated using a special sequencing method. Data on anatomical, structural, functional aspects and material properties were obtained from measurements and published data. The materials were idealized as transversely isotropic. The complete model consisted of 258 solid elements and 1635 nodes. RESULTS: The model was solved for displacements and stresses during clenching. In general, the observed displacement and stresses (tensile and compressive) were highest around the condylar region. Compressive stress was also observed around the premolar and molar bite points. CONCLUSION: This investigation has shown that the use of computer-aided modelling in conjunction with the finite element analysis could be effectively utilized in biomechanical analysis of the mandible. It could help to investigate many functional problems and could reduce the time of extensive experimentations. PMID- 15881306 TI - Open surgical approach for a tooth aspirated during dental extraction: a case report. AB - Foreign body aspiration is a common problem in children necessitating prompt recognition and early treatment to minimize the potentially serious and sometimes even fatal consequences. Most foreign bodies can usually be extracted by skilful application of endoscopic techniques. Nevertheless, spherical foreign bodies, such as pen caps and some teeth remain difficult to manage. In this case report clinical presentation and treatment options in the treatment of a patient who had a tooth lodged in the right lung are discussed. The complications of foreign body ingestion or aspiration associated with dental treatment is emphasized. PMID- 15881307 TI - Service provision in private general practice: 1983-1984 to 1998-1999. PMID- 15881308 TI - Changes in South Australian children's caries experience (continued from Aust Dent J 2004;49:212). PMID- 15881309 TI - Bacterial vaginosis and intravaginal practices: association with HIV. AB - OBJECTIVES: To determine the prevalence of bacterial vaginosis and to identify risk factors associated with this condition among urban women in Harare, Zimbabwe. DESIGN: A cross sectional study. MAIN OUTCOME MEASURES: Prevalent bacteria vaginosis (BV), HIV and intravaginal practices. SETTING: Urban primary health care clinics in Harare. SUBJECTS: 177 pregnant and 212 non pregnant women attending mother and child health care clinics. INTERVENTIONS: The women were recruited, counselled, interviewed, examined and tested for reproductive tract infections including HIV. Those with reproductive tract infections were given free treatment. RESULTS: The prevalence of bacterial vaginosis was 36% among HIV seropositive women and 26% among those seronegative. Fifty one per cent of the women practiced intravaginal cleansing, while another 28% reported use of intravaginal herbs. Women reporting intravaginal cleansing and/or use of intravaginal herbs were more likely to have bacterial vaginosis (OR 6.2 CI 3.6 to 10.7) and (OR 1.5 CI 1.1 to 2.5) and to be HIV seropositive (OR 1.8 CI 1.2 to 2.8) and (OR 1.8 CI 1.1 to 2.9) respectively. Other factors associated with bacterial vaginosis were malodorous vaginal discharge (OR 5.8 CI 2.9 to 10.7), genital warts (OR CI 3 CI 1.1 to 10.1) and Trichomonas vaginalis (OR 25.5 CI 11.6 to 56.7). CONCLUSIONS: BV was shown to be a common condition among the women the majority of whom indulged in intravaginal practices. HIV infection among women with BV and those that practiced intravaginal cleansing and/or use of herbs was significantly higher. Although causal relationship could not be established in this cross sectional study the results suggest that BV may facilitate HIV infection and we suggest the inclusion of information regarding risks associated with intravaginal practices into health education information disseminated to women. PMID- 15881310 TI - Adverse effects associated with the use of South African traditional folk remedies. AB - Although the toxicity of traditional folk remedies is well known in Africa, it is a subject which is surrounded by secrecy and has not been comprehensively studied. OBJECTIVES: The aims of this study are to describe the clinical features of patients admitted to hospital with a confirmed history of using folk remedies, and to gather data on their toxicity in a systematic fashion. DESIGN: Prospective case series. SETTING: Paediatric and adult wards of academic hospitals in Johannesburg, South Africa. SUBJECTS: The study population included 103 patients ranging from one day to 75 years of age, all of whom had recent folk remedy use. MAIN OUTCOME MEASURES: All available clinical data were analysed. Primary outcomes were the presence of renal and liver dysfunction, death or discharge from hospital. RESULTS: The most common clinical features on presentation were dehydration (51%), vomiting (46%), jaundice (40%), diarrhoea (39%), altered mental status (37%) and oligoanuria (30%). Renal dysfunction was present in 76% of patients and liver dysfunction in 48%. The overall mortality was 34%. The odds ratio of death was 5.1 (95% CI 1.41 to 18.5) in patients with renal dysfunction (p = 0.0077) and 5.35 (95% CI 1.99 to 14.4) in patients with liver dysfunction (p = 0.0006). CONCLUSION: Renal and liver dysfunction are frequently associated with use of folk remedies, and mortality in these patients is high. In view of the large numbers of African individuals living in the United States and Europe, it is important for physicians elsewhere to be aware of the potential toxicity of African folk remedies, and to inquire about their use. PMID- 15881311 TI - Cryptosporidiosis in Harare, Zimbabwe. AB - OBJECTIVE: To determine the prevalence of Cryptosporidium parvum in diarrhoeal patients. DESIGN: This was a laboratory-based cross sectional study on cryptosporidiosis in diarrhoeal patients. SETTING: Department of Medical Laboratory Sciences, College of Health Sciences in Harare, Zimbabwe. SUBJECTS: People of all ages with diarrhoea presenting at primary level health centres in Harare. MAIN OUTCOME MEASURES: Patient's age, laboratory results. RESULTS: Cryptosporidium parvum was the commonest enteric pathogen and was detected in 5.8% of the 500 diarrhoeal patients of all ages followed by Shigella species (3.8%) and Salmonella species (2.0%). The highest detection rate of C. parvum oocysts was observed in children less than five years old (11.2%) followed by children between six and 10 years old (6.3%) and then the 31 to 40 year age group (5.9%). CONCLUSIONS: Cryptosporidiosis affects people of all ages in Harare but is more common in children, particularly those under five years. The C. parvum oocysts should be looked for routinely in diarrhoeal stool specimens particularly those from children less than five years since C. parvum may be one of the causative agents of diarrhoea in this age group. PMID- 15881312 TI - Primary hypothyroidism and a concomitant pituitary mass: adenoma or pseudotumour? PMID- 15881313 TI - Vaginal hysterectomy. A five year prospective descriptive study. AB - OBJECTIVE: To review patients who had vaginal hysterectomy and observe any complications related to the procedure. DESIGN: A prospective descriptive study. SETTING: Sunshine Hospital, Actonville, Benoni, Gauteng Province, South Africa. SUBJECTS: 124 consecutive women who had vaginal hysterectomy. INTERVENTIONS: Vaginal hysterectomy. MAIN OUTCOME MEASURES: Duration of operation, hospital stay, intra and post operative complications; need for blood transfusion; histological result of uterus and uterus weight. RESULTS: The mean age was 44.7years (range 16 to 74 years). Mean parity was 2.7 (range 0 to 9). Common indications were menorrhagia (34.7%) and myomatous uterus (25.8%). The common risk factors were previous Caesarean delivery (12.9%), medical conditions (9.7%) and previous tubal ligation (10.5%). The mean operation time was 86.6 minutes (range 52 to 140 minutes). Only two (1.6%) patients required blood transfusion. Complications occurred in 5.6% of the patients. Myomatous uterus (40.3%) and adenomyosis (19.4%) were the commonest histological diagnosis. The mean hospital stay was 2.5 days (range 2 to 21 days). The average weight of the uterus was 142.2 gms (range 25 to 599gms). CONCLUSIONS: Vaginal hysterectomy can be performed and achieve very low complication rates. PMID- 15881314 TI - Pattern of cleft lip and palate [corrected] in Benin City, Nigeria. AB - OBJECTIVE: To study the pattern of cleft lip and/or palate (CL/P) in Benin City, Nigeria and to compare the findings with reports from other parts of the world. The results of this study may be used to improve the welfare of affected patients. DESIGN: A descriptive study. SETTING: The University of Benin Teaching Hospital (UBTH), Benin City, Edo State, Nigeria. SUBJECTS: 103 ethnic Nigerian patients with cleft lip and/or palate. MAIN OUTCOME MEASURES: Consecutive ethnic Nigerian patients presenting with CL/P were studied. Patients' data collected included age, sex, ethnic group, type of CL/P and the laterality pattern. Descriptive statistics were generated for all the variables recorded. RESULTS: There were 49.5% females and 50.5 % males; 95.1% were children and 4.9% were adults. The ethnic groups most often affected were Urhobo 29.1%, Bini 27.1% and Ibo 17.4%. The combined cleft lip and palate 60.2% was the commonest type of cleft. There were 28.2% patients with cleft lip only. More (7.8%) females were affected by the isolated cleft palate deformity. Twenty five patients had exceeded the time of repair. Their reasons were ignorance (36%) and financial constraints (64%) CONCLUSION: The data from this study is consistent with studies from other parts of the world. However, a high proportion of patients did not have access to early treatment due to ignorance and financial constraints. Public enlightenment and financial assistance for the indigent patient is recommended. PMID- 15881315 TI - Traumatic asphyxia during stadium stampede. AB - OBJECTIVES: To present a series of cases of survivors and non-survivors of traumatic asphyxia from a single mass casualty incident in Zimbabwe and a review of the literature. DESIGN: Descriptive case review. SETTING: Parirenyatwa Hospital is a tertiary referral 1 000 bed teaching hospital in Zimbabwe. RESULTS: Survivors (n = 4) displayed the classic signs of traumatic asphyxia of conjunctival haemmorhages, petechial blue-purple discoloration of head and neck and neurological findings of confusion or unconsciousness and convulsions. Non survivors (n = 12) showed more varied signs but all showed petechiae and with a history of being crushed. On-site resuscitation and triage was absent, reducing the chance of identifying potential survivors at the scene. CONCLUSION: The outcome in traumatic asphyxia is improved by rapid restoration of ventilation and circulation. The epidemiology of traumatic asphyxia in Zimbabwe is unknown but the conditions predisposing to it are present. Closer integration between hospital and pre-hospital services will permit better management of major trauma patients and mass casualty events. PMID- 15881316 TI - Forgotten diseases: relapsing fever. PMID- 15881317 TI - 5-HT2 receptor mediated the potentiation of GABA-activated current in the membrane of the dorsal root ganglion neurons of rat. AB - AIM: To explore the modulation of 5-HT on GABA-activated current (I(GABA)) in the membrane of rat dorsal root ganglion (DRG) neurons and its mechanism. METHODS: Rat DRG neurons were isolated mechanically and enzymatically, on which whole-cell patch clamp recording and repatch technique for intracellular dialysis were performed. RESULTS: In the majority of neurons examined (92.0%, 69/75) GABA induced a concentration-dependent inward current. In neurons sensitive to GABA preapplication of 5-HT produced potentiation effect (82.6% , 57/69) on I(GABA). Preapplication of 5-HT at concentrations of 1 x 10(-6), 1 x 10(-5), 1 x 10(-4) and 1 x 10(-3) mol x L(-1) potentiated I(GABA) by (35 +/- 8)% (n=8), (47 +/- 11)% (n=10), (65 +/- 17)% (n=9) and (75 +/- 18)% (n=11), respectively. This effect was mimicked by alpha-methyl-5-HT (1 x 10(-6) mol x L(-1)), a specific 5-HT2 receptor agonist, and reversed by cyproheptadine, a selective 5-HT2 receptor antagonist. The potentiation of I(GABA) by 5-HT was irrespective to whether the I(5-HT) presents or not in a subset of neurons. The concentration-response curves for GABA before and after pretreatment with 5-HT manifested the same threshold value and similar EC50 (2.0 x 10(-5) and 1.9 x 10(-5) mol x L(-1), respectively) , while the maximal value of I(GABA) for the latter was 33.6% higher than that for the former. Intracellular dialysis with GDP-beta-S or H-7 abolished the potentiation of I(GABA) by 5-HT, while H-9 did not. CONCLUSION: 5-HT can potentiate GABA-activated current via PKC-dependent phosphorylation of GABA(A) receptor following the activation of 5-HT2 receptor. PMID- 15881318 TI - M3-R/IK(M3)--a new target of antiarrhythmic agents. AB - AIM: To investigate the relationship between M3-R/IK(M3) and arrhythmia in order to find a new target for antiarrhythmic agents. METHODS: Using the acute ischemic model of rats and patch-clamp techniques, the effects of the M3 receptor on the occurrence of arrhythmias and its possible mechanisms were studied. RESULTS: In acute ischemic model of rats, the M3 receptor antagonist 4-diphenylacetoxy-N methylpiperidine-methiodide (4DAMP) increased the occurrence of arrhythmias, and the M3 receptor agonist choline suppressed the onset and the development of arrhythmias (P < 0. 01). No change was observed after treatment with other receptor antagonists (M1, M2, and M4). With patch-clamp techniques, it was found that choline induced K+ current could be inhibited by 4DAMP. Antagonists toward M1, M2, and M4 receptors all failed to alter the current. CONCLUSION: Choline modulates the cellular electrical properties of the heart, probably by activating a K+ current via stimulation of the M3 receptor. M3-R/IK(M3) may act as a new target for antiarrhythmic agents. PMID- 15881319 TI - Effect of 3, 4-dihydroxyacetophenone on Na+, K+ -ATPase activity of injured mitochondria and the oxygen consumption of brain cells of rat. AB - AIM: To investigate the effect of 3,4-dihydroxyacetophenone (alpha-DHAP) on Na+, K+ -ATPase activity of injured brain mitochondria induced by ascorbate-FeSO4 and the oxygen consumption of rat brain cells stimulated by ADP. METHODS: Na+, K+ ATPase activity was determined according to the method of inorganic phosphate. Swelling of the brain mitochondria was detected with the method of spectrophotometer. Lipid peroxidation was detected according to the thiobarbituric acid method of spectrophotometer. Oxygen consumption was measured by oxygen electrode method. RESULTS: The decrease of Na+, K+ -ATPase activity, mitochondria swelling and formation of lipid peroxidation were shown in rat brain mitochondria and cells induced by ascorbate-FeSO4. alpha-DHAP was shown to increase the activity of Na+, K+ -ATPase, decrease the mitochondria swelling and inhibit the production of lipid peroxidation of brain mitochondria and cells induced by ascorbate and FeSO4. alpha-DHAP can also reduce the oxygen consumption of brain cells stimulated by ADP. CONCLUSION: alpha-DHAP can protect the structure and the function of brain mitochondria and cells by scavenging the free radical and resisting the reaction of lipid peroxidation. PMID- 15881320 TI - Effect of rhIL-2 in inhalant form on experimental respiratory tract infection with Klebsiella pneumoniae in mice. AB - AIM: To determine the protective effect of recombinant human interleukin-2 (rhIL 2) in inhalant form on experimental respiratory tract infection with Klebsiella pneumoniae in mice. METHODS: Mice were infected with the method of nasal intubation drip. During infection, mice were given rhIL-2 by sc injection and the method of nasal intubation drip. There were normal group, vehicle group, model group, rhIL-2 groups and gentamicin group. In the end, the pathological changes in the lung were observed. The survival time and the mortality within a week of each group were recorded. The total protein content, the albumin content, the activity of alkaline phosphatase and the activity of lactic dehydrogenase of broncho-alveolar lavage fluid (BALF) were dertermined and compared. RESULTS: Symptoms of Klebsiella pneumoniae were remarkably relieved because of rhIL-2 administration. The total protein content, the albumin content, the activity of alkaline phosphatase and the activity of lactic dehydrogenase of BALF were less than those in the vehicle group and the model group. CONCLUSION: Inhalation of rhIL-2 can alleviate the pathological changes in the lung after infection. At the same dose, it could be seen that the effect of rhIL-2 in inhalant form was better than that of the injection. PMID- 15881321 TI - [Apoptosis of human carcinoma of mouth floor KB cells and multidrug resistant KBv200 cells induced by azide methyl anthraquinone derivative]. AB - AIM: To determine the effects of azide methyl anthraquinone derivative (AMAD) on growth inhibition and inducing apoptosis of multidrug resistant (MDR) KBv200 cells and parental drug-sensitive KB cells. METHODS: Cytotoxicity was determined by tetrazolium (MTF) assay. Reactive oxygen species (ROS) levels and mitochondrial membrane potential (deltapsi(m)) in cells were labeled with DCFH-DA and DiOC6 and tested by flow cytometry. Annexin V stain and DNA ladder were used to examine the apoptosis of KB and KBv200 cells induced by AMAD. RESULTS: AMAD was shown to inhibit the growth of KB and KBv200 cells significantly in a concentration-dependent manner, with mean IC50 of 0.36 and 0.45 micromol x L(-1), respectively. The generation of ROS increased obviously after the cells were treated with AMAD for 12 h, up to the peak in 24 h, meanwhile the levels of deltapsi(m) were time-dependently decreased. DNA fragmentation appeared on the agarose gel. Annexin V stain showed AMAD induced apoptosis of KB and KBv200 cells also in a concentration-dependent manner. CONCLUSION: AMAD showed inhibitory effect on both MDR KBv200 cells and parental drug-sensitive KB cells. The mechanism of action was associated with the increase of the cellular ROS level and the decrease of the mitochondrial membrane potential induced by AMAD, which result in cell apoptosis. PMID- 15881322 TI - [Vitexicarpin, a flavonoid from Vitex trifolia L., induces apoptosis in K562 cells via mitochondria-controlled apoptotic pathway]. AB - AIM: To investigate the inhibitory effect of vitexicarpin on the proliferation of human cancer cells and its mechanism of action. METHODS: The inhibitory effect of vitexicarpin on the proliferation of human cancer cells was evaluated by the SRB method and its apoptosis-inducing effect was demonstrated by morphological observation under light microscope, flow cytometric analysis and agarose gel electrophoresis. The proteins related to apoptosis were examined by Western blotting analysis. RESULTS: Vitexicarpin significantly inhibited the proliferation of human cancer cells, A2780, HCT-15, HT-1080 and K562, with the IC50 values of (19.1 +/- 2.4) micromol x L(-1) for A2780(48 h), (0.66 +/- 0.10) micromol x L(-1) for HCT-15(48 h), (0.44 +/- 0.06) micromol x L(-1) for HT-1080 (48 h) and (0.28 +/- 0.14) micromol x L(-1) for K562 (24 h). The cells treated with vitexicarpin showed characteristic morphology typical for apoptosis and gave dose-dependent sub-G0/G1 peak in the flow cytometric analysis and DNA ladder on agarose gel electrophoresis. In Western blotting analysis, the cleavage of PARP and caspase-3, the release of cytochrome c from mitochondria into the cytosol, the decrease of Bcl-2 expression level, and the down-regulation of the ratio of Bcl-2/Bax expression level were examined in the K562 cells treated with vitexicarpin. CONCLUSION: Vitexicarpin induces apoptosis in K562 cells via mitochondria-controlled apoptotic pathway. PMID- 15881323 TI - [Physical-chemical properties and structure elucidation of abPS isolated from the root of Achyranthes bidentata]. AB - AIM: To study the physicochemical properties and the structure of Achyranthes bidentata polysaccharide (AbPS). METHODS: AbPS was isolated from the roots of Achyranthes bidentata Bl., and purified by gel filtration chromatography. The distribution of the molecular weight of AbPS was determined by ESI-MS. The structure of AbPS was deduced by methylation analysis, reductive-cleavage and 13CNMR spectroscopy. RESULTS: AbPS was shown to compose of fructose residues and glucose residues and the molar ratio was 8:1. AbPS contain 2,1-linked fructose residue, 2,1-linked fructose residue, 1,2,6-linked fructose residue, terminal fructose residue and terminal glucose residue. CONCLUSION: AbPS is a fructan and belong to graminan. PMID- 15881324 TI - A new picfeltarraenone glycoside from Picria fel-terrae. AB - AIM: To investigate the chemical constituents from Picria fel-terrae Lour. METHODS: Column chromatography techniques were used to isolate the chemical constituents, physico-chemical constants and spectroscopic analysis were employed for structural elucidation. Results Two triterpenoids named picfeltarraenone I (1) and picfeltarraenin XI (2) were isolated, and their structures were established to be 3,11,22-trioxo-16alpha-hydroxy-(20S,24)-epoxy-cucurbit-5,23 diene (1) and 3,11,22-trioxo-16alpha-hydroxy-(20S,24)-epoxy-cucurbit-5, 23-diene 2beta-O-beta-D-glucopyranoside (2), respectively. CONCLUSION: Compound 2 is a new compound, the 13CNMR data of compound 1 is reported for the first PMID- 15881325 TI - [Liquid chromatography frontal analysis of the protein binding of glimepiride]. AB - AIM: To study the protein binding of glimepiride. METHODS: An HPLC-FA method is performed by using Pinkerton GFF II-S5-80 internal-surface reversed-phase silica support (150 mm x 4.6 mm ID, 5 microm) at pH 7.4 in a 67 mmol x L(-1) isotonic sodium phosphate buffer at 37 degree C. Other conditions included flow rate of 0.2 mL x min(-1), UV detection at wavelength 230 nm and injection volume 900 microL. RESULTS: Nonlinear regression parameter estimation was used for the association constant measurement of glimepiride to both primary and secondary sites, which were 5.1 (micromol x L(-1)-1 and 1 for K1 and n1, and 0.017 (micromol x L(-1))-1 and 7 for K2 and n2, respectively. CONCLUSION: The method is shown to be suitable for investigation of protein binding of glimepiride. PMID- 15881326 TI - Monosaccharide composition analysis of glycoproteins by isotope-tag method and capillary LC/ESI-MS. AB - AIM: To develop a rapid and sensitive method for monosaccharide composition analysis. METHODS: Glycoprotein was first hydrolyzed to monosaccharides, which were subsequently reacetylated (amino monosaccharides), tagged with 2 aminopyridine and then separated and monitored in selected ion mode by CapGCC LC/MS. The use of tetradeuterium labeled-aminopyridyl-monosaccharides prepared by tagging monosacahrides with hexadeuterium labeled 2-aminopyridine as internal standards improved the linearity and reproducibility in quantification. RESULTS: This method was successfully applied to monosaccharide composition analysis of model glycoproteins, fetuin and erythropoietin down to 1 pmol monosaccharides. CONCLUSION: This method has been shown to be highly sensitive and is applicable to monosaccharide composition analysis of glycoproteins. PMID- 15881327 TI - [Evaluation of measurement uncertainty for the determination of ginsenosides in Radix ginseng by HPLC]. AB - AIM: To set out the procedure for estimation of measurement uncertainty for the determination of ginsenosides R(g1), Re and R(b1) in Radix ginseng by HPLC. METHODS: To facilitate the identification and analysis of the uncertainty sources arising from the procedure of analysis, a cause and effect diagram was constructed and simplified. Each uncertainty component whether associated with individual sources or with the combined effects of several sources, was evaluated with respect to the significance of its contribution to the overall measurement uncertainty and was expressed as standard uncertainty. All the standard uncertainties were then combined according to the appropriate rules to give a combined standard uncertainty and an expanded standard uncertainty. Results The expanded standard uncertainties for the HPLC determination of ginsenoside R(g1), Re, and R(b1), are 0.12c, 0.14c and 0.13c, respectively. CONCLUSION: Measurement uncertainty is applicable to set the limit of the ginsenosides in Radix ginseng. The establishment of the methodology for the evaluation of measurement uncertainty is important to the studies of Chinese materia medica standards. PMID- 15881328 TI - [Excretion of beta-elemene from rat respiratory tracts]. AB - AIM: To investigate the excretion of beta-elemene from the respiratory tracts in male Spraque-Dawley rats. METHODS: After a single administration of beta-elemene to rats at the dosage of 75 mg x kg(-1) (i.v. or i.p.), the exhaled gases were collected and concentrated at various time points. The residues were analyzed by gas chromatography. RESULTS: A minor amount of unchanged beta-elemene was excreted via rat respiratory tracts after iv and ip administration of a single dose. The cumulative excretion were 1.41% and 0.51% respectively. CONCLUSION: The results demonstrated that unchanged beta-elemene excretes from rat respiratory tracts, but may not be the main elimination pathway in rats. PMID- 15881329 TI - The stability of insulin-loaded polybutylcyanoacrylate nanoparticles in an oily medium and the hypoglycemic effect in diabetic rats. AB - AIM: To study the stability of insulin-loaded polybutylcyanoacrylate nanoparticles (IPN) in an oily medium (soybean oil containing 0.5% (v/v) Tween-20 and 5% (v/v) Vitamin E) along with the hypoglycemic effect following their oral administration to streptozotocin (STZ) induced diabetic rats. METHODS: The stability of IPN in the process was appraised by measurement of the amount of undegraded insulin associated to nanoparticles, the average size and the span of IPN, as well as the release of insulin from IPN. IPN in an aqueous medium (containing 0.5% (v/v) Tween-20) at pH 2.0 was also investigated as control. RESULTS: The study showed that IPN in the oily medium was more stable than that in the aqueous medium over one year of storage in the dark at (25 +/- 2) degrees C and the in vitro stability of IPN in the oily medium against degradation by proteolytic enzymes was much better than that in the aqueous medium. The apparent bioavailability of an oral administration of IPN (50 u x kg(-1)) in the oily medium versus an (sc) injection of insulin (2 u x kg(-1)) was 22.4%, much higher than that of IPN in the aqueous medium (15.5%), based on decreased areas above curve (AAC) determination for the blood glucose depression from time zero to 144 h of a single oral administration of IPN to STZ-diabetic rats. CONCLUSION: IPN in soybean oil containing Tween-20 (0.5% v/v) and Vitamin E (5% v/v) could be considered as an effective and stable delivery system for oral insulin. PMID- 15881330 TI - [Mechanisms of action of transportation of liposomes and chitosan-coated liposomes containing leuprolide across intestine and Caco-2 cell]. AB - AIM: To investigate the mechanisms of action of transportation of liposomes and chitosan-coated liposomes containing leuprolide across rat intestine and Caco-2 cell. METHODS: Everted-gut technique and Caco-2 cell were used to study the transport properties of free leuprolide, liposomes and chitosan-coated liposomes containing leuprolide. Caco-2 cell was used to study the effect of chitosan concentration and the order of addition on the permeation of liposomes. RESULTS: The transport of leuprolide was passive diffusion. Probably because the entrapment by liposomes prevents the transport of leuprolide across the rat intestine and Caco-2 cell, the permeation amount of leuprolide from liposomes was lower than that of the free drug. However, liposomes protected the leuprolide from degradation. Chitosan promoted the transport of leuprolide from liposomes and there was no obvious difference in enhancement effect from the concentration of 0.1% to 0.5%. On the other hand, the incubation of chitosan with liposomes may weak the enhancement effect of chitosan. CONCLUSION: Chitosan-coated liposomes showed both protection and enhancement effect, therefore, they may promote the oral absorption of leuprolide. PMID- 15881331 TI - [Assesement of tretinoin with a self-emulsifying formulation in vitro and in vivo]. AB - AIM: To evaluate the self-emulsifying ability and dissolution behavior of tretinoin in vitro and the pharmacokinetic behavior in beagle dogs. METHODS: The self-emulsifying rate was evaluated by determining the intensity of scattered light at different time and the particle size of resultant emulsions after self emulsifying were observed by optical microscope. The plasma concentrations were determined by HPLC and dissolution and pharmacokinetic behavior of self emulsifying formulations were evaluated by comparison with commercial capsules. RESULTS: The area under the curve (AUC) was significantly higher in the tretinoin self-emulsifying formulation group (3048.0 mg x h x L(-1)) than that in the commercial capsule group (1826.0 mg x h x L(-1)). Also, Tmax was smaller in the self-emulsifying formulation group (1.25 h) compared with market products (2 h) and the dissolved amount from self-emulsifying formulations in water at 15 min was much higher (more than 80%) than that of the market products (less than 5%). CONCLUSION: The self-emulsifying drug delivery systems can increase drug dissolution in vitro and absorption in vivo significantly. PMID- 15881332 TI - [Authentication of Zanthexylum bungeanum Maxin population and adulterants by analysis of rDNA ITS sequences]. AB - AIM: To study the difference of rDNA ITS sequences between Zanthexylum bungeanum populations and their adulterants in main habitants of China so as to provide molecular markers for identifying Zanthexylum bungeanum populations against adulterants. METHODS: rDNA ITS regions (including ITS-1, 5.8S and ITS-2) of 7 populations of Zanthexylum bungeanum which are separate located in Gansu, Shanxi, Sichuan, Hebei provinces, and 3 adulterants were sequenced by PCR products sequencing method or clone sequencing method. RESULTS: The sequences of rDNA ITS region of Zanthexylum bungeanum were reported for the first time, and the sequences of ITS region ranged from 619 to 620 bp, and the length difference amoung Zanthexylum bungeanum and their adulterants is 4 bp. There are 15 variable sites, 12 informative sites and 3 authenticable sites among Zanthexylum bungeanum populations. The difference of rDNA ITS regions amoung Zanthexylum bungeanum and their adulterants is obvious, the number of variable sites is 71. CONCLUSION: The difference of rDNA ITS sequences can be used to authenticate accurately the populations of Zanthexylum bungeanum and their adulterants. These populations of Z. bungeanum which have close relationship always distribute in near geographic areas. The characteristics of rDNA ITS sequence can be used as good markers for authenticating Zanthexylum bungeanum populations form their adulterants. PMID- 15881333 TI - [In intro delivery of gene encoding neurotrophic (GDNF) into brain by gene transfer]. AB - AIM: The gene encoding neurotrophic factor was transfected into brain capillary endothelial cells with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs. METHODS: Plasmid DNA encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was constructed and prepared. Then, mGDNF gene was transfected into cultured mouse brain capillary endothelial cells (BCECs) in vitro. The amount of mGDNF protein in the transfected cells and secreted from the transfected cells were determined by ELISA. The polarity of the secretion of mGDNF protein from BCECs was investigated in a bicameral culture system. RESULTS: The mGDNF protein was detected out not only from the transfected cells but also the cultured media. And mGDNF protein was mainly found in the brain side of the culture compartment. CONCLUSION: It has been demonstrated that a secretory protein can be successfully delivered into brain parenchyma by utilizing the secretory pathway of BCECs. PMID- 15881334 TI - [Targeted AGEes and AGEs cross-link in drug discovery: preventing and reversing arterial sclerosis in aging and diabetes]. PMID- 15881335 TI - [Grasps favorable opportunity to search for truth]. PMID- 15881336 TI - [PPAR family and its relationship to metabolic syndrome]. AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand--activated transcription factors. Three PPAR isoforms , designated PPARalpha, -beta/delta, and -gamma, have been identified and attracted enormous attention due to the key role these receptors play in regulating adipogenesis, lipid metabolism, insulin sensitivity, inflammation and blood pressure. Growing evidence points to a causative relationship between PPAR activity and the metabolic syndrome, including insulin resistance, glucose intolerance or type II diabetes, obesity, dyslipidemia, hypertension, atherosclerosis, and albuminuria. Importantly, both PPARalpha activators such as fibric acid class of hypolipidemic drugs and PPARgamma agonists including antidiabetic thiazolidinediones (TZDs) have been proved to be effective for improving metabolic syndrome. All three PPAR isoforms appear to play important roles in the development of type II diabetes and diabetic nephropathy. Accumulating data has begun to emerge suggesting PPARs may serve as potential therapeutic targets for treating the metabolic syndrome and its related complications. Here we review the literature pertaining to the action, ligand selectivity and physiological role of PPARs. Particular emphasis is placed on their pathogenic roles in the metabolic syndrome and the therapeutic utility of PPAR modulators in the treatment of type II diabetes. PMID- 15881337 TI - [The study of the mechanisms underlying the effects of general anesthetics on spinal cord]. AB - There are very extensive and complex effects of general anesthetics on spinal cord, so the advanced study on the mechanisms underlying the effects of general anesthetics at the spinal cord level will undoubtedly promote the more reasonable application of the general anesthetics and the understanding of mechanisms of general anesthesia. This paper reviews the actions of general anesthetics on neurons, synaptic transmissions, related receptors as well as ion channels in the spinal cord. PMID- 15881338 TI - [Progress in the study of ryanodine receptor-related muscle diseases]. AB - Ryanodine receptor (RyR), a calcium release channel within the sarcoplasmic reticulum (SR) membrane, is a key protein in skeletal and cardiac excitation contraction coupling. The structural and functional changes of RyR can result in decoupling of excitation-contraction, and lead to muscle diseases. Current study confirms that RyR is related to the pathogenesis of these diseases. In this review, we look at the current progress in the study on RyR and these diseases to provide the molecular basis in the prevention and therapy of these diseases. PMID- 15881339 TI - [Involvement of N-type calcium channels in pain and antinociception]. AB - N-type voltage-dependent calcium channels are critical for pain transduction and modulation. These channels are highly present at the presynaptic terminals of nociceptive neurons in dorsal horn of the spinal cord where they regulate the release of the key pro-nociceptive neurotransmitters such as glutamate and substance P. Consistent with this, the selective blockers of the N-type channels are powerful analgesics, and, mice lacking the N-type Ca(v)2. 2 subunit have higher pain thresholds compared to wild-type. The N-type channels are also present at synapses in the autonomic system and the central nervous system, the inhibition of these synapses is the main reason of the side effects in analgesic therapy with current N-type channel blockers. Recently, a unique splice isoform of the N-type channels was found exclusively expressed at the nociceptive neurons in dorsal root ganglia, which may represent a novel target for pain management. PMID- 15881340 TI - [Sterol regulatory element-binding proteins and lipid metabolism]. AB - Lipid homeostasis in vertebrate cells is regulated by a family of membrane-bound transcription factors designated sterol regulatory element-binding proteins (SREBPs). SREBPs directly activate the expression of numerous genes dedicated to the synthesis and uptake of cholesterol, fatty acids, triglycerides, and phospholipids, together with the NADPH cofactor required to synthesize these molecules. In this review, we focus on the mechanism of SREBP trafficking and processing, as well as the molecular aspects of SREBPs in regulating cellular lipid homeostasis; furthermore, we discuss the involvement in the diseases relevant to the disturbance of lipid metabolism. PMID- 15881341 TI - [The advances in basal research of SARS CoV]. PMID- 15881342 TI - [The inhibiting effect of uncoupling protein 2 on the production of reactive oxygen species]. PMID- 15881343 TI - [A novel member of endothelin family--endothelins1-31]. PMID- 15881344 TI - [Advances in the study of preganglionic cardiac parasympathetic neurons]. PMID- 15881345 TI - [bHLH transcription factors in the development of neocortex]. PMID- 15881346 TI - [Protective role of neutral endopeptidase in cardiovascular diseases]. PMID- 15881347 TI - [Progress of the angiogenic mechanism of the angiogenin and the relationship between angiogenin and diseases]. PMID- 15881348 TI - [CIC CI- channels]. PMID- 15881349 TI - [The role of voltage-gated sodium channels in neuropathic pain]. PMID- 15881350 TI - [Cholesterol metabolic disorders and Alzheimer's disease]. PMID- 15881351 TI - [Inflammatory pathology and the anti-inflammation of saccharide substances in Alzheimer's disease]. PMID- 15881352 TI - [Loss of IGF2 imprinting in colorectal cancer]. PMID- 15881353 TI - [Role of alpha-tocopherol and L-ascorbic acid in mammalian reproduction]. PMID- 15881354 TI - [Molecular basis of sperm-egg membrane adhesion and fusion in mammals]. PMID- 15881355 TI - [The development of TRP family relating to thermosensation]. PMID- 15881356 TI - [Application of nonlinear dynamics in EEG analysis]. PMID- 15881357 TI - [Molecular topology study of gas chromatographic retention indices of alkane series]. AB - The gas chromatographic retention indices can be used to qualify some organic compounds. A new topological index based on distance matrix and branch vertex of the atoms in a molecule is proposed by defining equilibrium electronegativity of atoms in the molecule and coloring atoms in the molecular graph with equilibrium electronegativity, which appears unique to the molecular structures and has excellent structural selectivity. The multivariate linear equations of gas chromatographic retention indices are as follows: I(Squalane) = 23.97842N1 - 3.86562N2 + 0.787379N3 + 42.33061, R = 0.9922, n = 70, S = 13.70405, F = 1396.601; I(SE-30) = 23.83937N1 - 3.5687N2 + 0.939876N3 + 22.11952, R = 0.9919, n = 37, S = 11.96088, F = 668.8781; where the N1, N2 and N3 are a group of topological indices; n, R, S and F are sample number, regression coefficient, residual standard deviation and F-statistic value, respectively. The calculated results by the formulae indicate that the average relative deviations between calculated values and experimental data of gas chromatographic retention indices of alkane series on both squalane (column temperature 50 degrees C) and SE-30 (column temperature 80 degrees C) were all 1.31% and the errors were within experimental deviations. The equations can express well the change rule of the relative gas chromatographic retention indices of alkane series. PMID- 15881358 TI - [Effect of synthesis conditions on the morphology and binding property of (-) ephedrine imprinted polymers]. AB - Contribution of the synthesis parameters to the properties of the polymer is one of the major interests in the study of molecular imprinting technology. To have a better understanding of the influence of polymerization conditions on the polymer morphology and binding property, (-)-ephedrine imprinted polymers synthesized with different cross-linkers (ethylene glycol dimethacrylate (EDMA) and pentaerythritol triacrylate (PETRA)), porogens (chloroform and acetonitrile) and monomer concentrations were compared. The resulting polymers were characterized with their pore structure, texture and binding affinities by BET and equilibrium adsorption method. The selectivity of the polymer was evaluated with chromatographic analysis. The results suggested that CHCl3 is a good solvent for methacrylic acid (MAA)-EDMA and MAA-PETRA polymer chains which resulted in smaller polymer surface area. Quantitative determinations of the carboxyl groups in the polymer indicated that polymers synthesized with the same pre polymerization composition except porogen can have different concentrations of functional monomer in their structures. The study showed that porogen in the molecular imprinting synthesis can affect the morphology and composition of the polymers, which influence the binding affinity of the polymers. PMID- 15881359 TI - [Separation of proteins in aqueous two-phase systems with high-speed counter current chromatography]. AB - High-speed counter-current chromatography (HSCCC) is a continuous liquid-liquid partition chromatography, with remarkable advantages of high separation efficiency and no adsorption or denaturation by solid phase. The retention of stationary phase and the separation of proteins in polyethylene glycol 1000 (PEG1000)-phosphate aqueous two-phase system (ATPs) were studied with a multi column high speed-counter-current chromatograph. The flow direction and speed of the mobile phase, and the rotation direction and speed of the apparatus showed different effects on the retention of the stationary phase, which reached the maximum at 33.3% with a flow rate of 0.6 mL/min and a rotation speed of 900 r/min in 14.0% PEG1000-16.0% phosphate ATPs. Distinct differences in partition coefficients among cytochrome C, lysozyme and hemoglobin were found at pH 9.2 and these three proteins were successfully separated in 14.0% PEG1000-16.0% phosphate ATPs at pH 9.2 by HSCCC with the apparatus rotating at 850 r/min and the mobile phase flow rate of 1.0 mL/min. The major protein components in hen egg white, including ovaltransferrin, ovalbumin and lysozyme also show distinct differences of partition coefficients in PEG1000-phosphate ATPs at pH 9.2. Ovalbumin and lysozyme were successfully purified to homogeneity and ovaltransferrin to ca 60% purity from the hen egg white sample with yields over 90% in 15.0% PEG1000-17.0% phosphate ATPs at pH 9.2 with the apparatus rotating at 850 r/min and mobile phase flow rate of 1.0 mL/min. PMID- 15881360 TI - [Determination of dopamine and norepinephrine in Portulaca oleracea L. by micellar electrokinetic capillary chromatography with amperometric detection]. AB - A method based on micellar electrokinetic capillary chromatography with electrochemical detection (MEKC-EC) was developed for the simultaneous determination of norepinephrine (NE) and dopamine (DA) in Chinese herbal plant extracts of Portulaca oleracea L. The effects of several factors such as the acidity and concentration of running buffer, sodium dodecylsulfate (SDS) concentration and detection potential were investigated to acquire the optimum conditions. The two analytes could be well separated within 12 min in a 64 cm capillary at the separation voltage of 25 kV in a 10 mmol/L phosphate buffer (pH 6.23) containing 10 mmol/L SDS. The excellent linearity was obtained in the concentration range from 1.0 x 10(-6) to 5.0 x 10(-4) mol/L. The detection limits of concentration were 4.2 x 10(-7) mol/L for NE and 8.7 x 10(-7) mol/L for DA and those of quantity were 0.41 fmol for NE and 1.45 fmol for DA at a signal-to-noise ratio of 3. This method was successfully used in the analysis of Portulaca oleracea L. without derivatization procedure, and real average contents of NE and DA in Portulaca oleracea L. were 0.015% and 0.20%, respectively. PMID- 15881361 TI - [Capillary electrophoresis with end-column electrochemical detection for hydrochlorothiazide and triamterene diuretics]. AB - A method based on capillary electrophoresis with end-column electrochemical detection (HPCE-ED) was developed for the determination of the diuretics of hydrochlorothiazide (HCT) and triamterene (TAT) simultaneously. The detection electrode was a 300 microm carbon disc electrode at a working potential of +1.1 V versus Ag/AgCl electrode. The two analytes could be well separated within 8 min in a 50 cm long capillary at a separation voltage of 24 kV with a 10 mmol/L phosphate buffer (pH 7.5). Under optimum conditions, the current response was linear over about two orders of magnitude with detection limits (S/N = 3) of 0.29 and 0.25 mg/L for triamterene and hydrochlorothiazide, respectively. The proposed method was successfully applied to determine the synthetic urine and real pharmaceuticals samples. The recoveries were found to be in the range of 93.5% 97.2%. PMID- 15881362 TI - [New advances of high performance separation and analytical techniques for proteins in complex biological systems]. AB - As a new strategy, proteomics is playing more and more important roles in the life sciences after the sequencing of the human genome was completed. Because a biological system is very complex and diverse, the study and application of technological platform for the separation and analysis with high resolution, high sensitivity, high throughput and broad dynamic range have become one of the important and hot spots. The review focused on the new progress of high performance separation and analysis techniques which were applied to the field of proteomics increasingly, such as multidimensional liquid chromatography for pre fractionation, capillary liquid chromatography coupled with mass spectrometry and capillary electrophoresis interfaced on line with mass spectrometry. PMID- 15881363 TI - [Multidimensional separations used in pharmaceutical and biological fields]. AB - A review of multidimensional separations such as comprehensive two-dimensional gas chromatography (GC x GC), comprehensive two-dimensional high performance liquid chromatography (HPLC x HPLC) and their applications in pharmaceutical and biological fields is presented with 71 references. A single CO2 cryo-jet loop modulator was developed for GC x GC and it can be used to modulate compounds higher than C6 effectively. Comprehensive two-dimensional gas chromatography-time of-flight mass spectrometry (GC x GC/TOF-MS) analyses of traditional Chinese medicine volatile oils such as Pogostemon cablin Benth (Cablin Patchouli), Forsythia suspensa (Thunb.) Vahl and Zedoary were reported also. As an emerging technology, multidimensional separations hold the promise and play an important role in the future pharmaceutical and biological fields. PMID- 15881364 TI - [Dead time determination of the second dimension in a comprehensive two dimensional gas chromatography]. AB - Comprehensive two-dimensional gas chromatography (GC x GC) is rapidly gaining importance for the analysis of complex samples. In order to predict chromatogram and optimize the operational conditions, the dead time of each dimension should be obtained exactly. Two methods to calculate the dead time of the second dimension were introduced. In the first method, based on the relationship between the time differences of a series of n-alkanes substances and the apparent retention times under different pressures, the true retention time can be calculated for non-synchronous GC x GC. The retention times of homologous series were used to calculate the dead time. Another rapid method to calculate the dead time of the second dimension is based on three or more apparent retention times in a single temperature programmed chromatographic run. The results show that the dead time of the second dimension can be calculated successfully by using the proposed methods and the deviation between the two methods is less than 0.05 s. PMID- 15881365 TI - [Nano flow multidimensional liquid chromatography for proteome analysis of rat liver]. AB - A nano flow comprehensive multidimensional liquid chromatography was developed. By using this separation platform, complex samples can be injected, desalted, separated and analyzed in complete automatization. Coupling of the strong cation exchange (SCX) with capillary reversed-phase liquid chromatography (cRPLC) is described for proteomics of rat liver. A step gradient was applied for SCX column, and a linear gradient was applied for RPLC. The flow rate through the nano scale analytical column during the separation process was maintained at 200 nL/min. The peak capacity for the nano-2D-LC was up to 620 under these conditions. PMID- 15881366 TI - [Applications of two-dimensional liquid chromatography coupled to mass spectrometry for the separation and identification of compounds in ginkgo biloba extracts]. AB - A comprehensive two-dimensional liquid chromatographic separation system based on the combination of an immobilized liposome chromatographic (ILC) column and an ODS column was developed for the separation of components in Ginkgo biloba, a traditional Chinese medicine. Two columns were coupled by a two-position, eight port valve equipped with two storage loops, and the system was controlled by a computer. The effluent was detected both by a diode array detector and by an atmospheric pressure chemical ionization (APCI) mass spectrometer. Under the optimization separation conditions with the separation system, more than 41 components in the methanol extract of Ginkgo biloba were resolved. According to their UV and mass spectra, 13 of them were preliminarily identified as ginkgolide B, ginkgolide C, bilobalide, rutin, quercetin, quercetin-3-O-beta-D-glucosyl (1 2)-alpha-L-rhamnoside, quercetin-3-O-beta-D-glucoside, isorhamnetin, kaempferol-3 O-beta-D-glucosyl (1-2)-alpha-L-rhamnoside, isohamnetin-3-O-beta-D-rutinoside, kaempferol-3-O-beta-D-glucoside, kaempferol, kaempferol-3-O-beta-D-rutinoside. PMID- 15881367 TI - [Application in capillary electrophoresis-based two-dimensional separation system]. AB - An on-column, etched fused-silica porous junction interface was designed and fabricated, and an integrated two-dimensional separation platform involving on line coupling of capillary isoelectric focusing (CIEF) with capillary electrophoresis (CE) was constructed by the interface. The primary advantages of this novel porous junction interface over previous designs are no dead volume, simplicity, ruggedness and convenience of column switching. This interface is particularly well suited for on-line coupling of CE-based multiple dimensional separation system. The feasibility and performance of the 2D CIEF-CE system constructed by such an etched porous junction were evaluated by the separation of soluble extracts of an antler. The results indicated that the total separation time was less than 1 h, and the 2D CE separation system was found to increase the resolving power and overall peak capacity over single dimension mode. PMID- 15881368 TI - [Determination of carotenoids in foods by high performance liquid chromatography]. AB - Recent research on the types and contents of carotenoids in food emphasis has been increasingly placed on obtaining more accurate data. The analysis of carotenoids, however, is challenging because of the diversity and the presence of cis-trans isomers, in addition to the characteristic conjugated double bond system of carotenoids causing their particular instability, especially under light, heat, oxygen and acids. The determination of carotenoids and carotenoid ester in foods using high performance liquid chromatographic methods are reviewed. In addition, the sample extraction, treatment and some methods for chromatographic separation and analysis are briefly commented on. PMID- 15881369 TI - [Advances of the coatings used in columns for capillary electrophoresis and in nanochannels of chips]. AB - An overview is provided on the advancement and development of coating preparation methodology and materials used in capillaries and channels in microfluidic chip. Discussion is also given on the effects of coatings in the resolutions of separation and the reproducibility of separations. Dynamic coatings and linked coatings, classified as homo-polymers, copolymers and heterocyclic compounds, are further discussed, and so are the methods for the preparation of the coatings by cross-linked reaction, sol-gel process, photomodification and chemical deposition, etc. The discussion will be useful for the optimization of capillary columns that are used in capillary electrophoresis and nanochannels of chip. PMID- 15881370 TI - [Study on the chromatographic behavior of bases on dodecylamine-N,N-dimethylene phosphonic acid modified zirconia stationary phase]. AB - A new zirconia-based stationary phase (DPZ) was prepared by modifying zirconia with dodecylamine-N,N-dimethylenephosphonic acid (DDPA). DDPA adsorbs on zirconia with only one phosphonic group, and another is free. Besides the hydrophobic interaction provided by non-polar dodecyl, DPZ also has dipolar interaction, ion exchange and electrostatic repellent interaction provided by the free phosphonic group and amino group at different conditions. Separations of bases on this stationary phase were achieved with satisfaction due to the various retention mechanisms. The influences of methanol content, pH value, ion type and ion strength of mobile phase were studied in detail. PMID- 15881371 TI - [Development of high performance liquid chromatographic method using monolithic column for the determination of urinary nucleosides]. AB - Modified nucleosides excreted in urine have been studied as possible bio-markers for malignant tumors. Reversed-phase high performance liquid chromatography (RP HPLC) is one of the most popular analytical tools. However, its analysis time is longer than desired. The analytical method for urinary nucleosides has been developed using a monolithic column and a linear gradient elution of 25 mmol/L KH2PO4 solution (pH 4.55) and 60% methanol in water with UV detection at 260 nm. Twelve urinary nucleosides were completely separated. The monolithic column presented similar analytical characteristics to reversed-phase column. The time of the analysis is only 23 min, in general much shorter than the methods using a packed RP-HPLC. The method developed is sensitive, reliable and suitable for clinical applications. PMID- 15881372 TI - [Measurement of monoamine transmitters in the brains of ovariectomized mice using high performance liquid chromatography with fluorescence detection and gradient flow rate elution]. AB - A method for the determination of norepinephrine (NE), dopamine (DA) and 5 hydroxytryptamine (5-HT) in brains of ovariectomized mice was established by using high performance liquid chromatograph with a fluorescence detector, a C18 column and a gradient flow rate delivered by a single pump. The mobile phase was a mixture of 0.1 mol/L KH2PO4 and CH3OH (9/1, v/v). Detection limits of the method were determined to be lower than 10 nmol/L, absolute recoveries were above 80% and the relative standard deviations (RSDs) of intra-day were all within 3.03%. The method has the advantages of high sensitivity and of simple sample preparation. Short analysis time and better separation of samples were also obtained. PMID- 15881373 TI - [Determination of angiotensin converting enzyme inhibitor activity by high performance liquid chromatography]. AB - A high performance liquid chromatographic method to determine angiotensin converting enzyme inhibitor activity in vitro was established by using N-hippuryl His-Leu tetrahydrate as the reaction substrate and hippuric acid as the reaction product. The chromatographic conditions were as follows: column, ZORBAX SB-C18 (4.6 mm i. d. x 150 mm, 5 microm); column temperature, 25 degrees C; mobile phase, acetonitrile-distilled water (25:75, v/v, both containing 0.05% (v/v) trifluoroacetic acid and 0.1% (v/v) triethylamine); flow rate, 0.5 mL/min; detection wavelength, 228 nm. An excellent linearity over the range of 0.005 1.000 mmol/L (r = 0.9999) was observed. The detection limit was 0.50 micromol/L. The recoveries of hippuric acid ware 99.48%-105.64%, with a relative standard deviation (RSD) of 2.20% (n = 6). It is a simple, precise and reliable assay method for developing antihypertension drugs. PMID- 15881374 TI - [Simultaneous determination of pseudoephedrine hydrochloride and dextromethorphan hydrobromide in composite pseudoephedrine hydrochloride dry suspension by HPLC]. AB - A simple and reliable reversed-phase high performance liquid chromatographic method (HPLC) for the routine analysis of pseudoephedrine hydrochloride and dextromethorphan hydrobromide in composite pseudoephedrine hydrochloride dry suspension has been established. HPLC determination for the drug was performed in a Lichrospher C6H6 column and detected at 220 nm. Acetonitrile-H2O-H3PO4 (50:50:0.1, v/v, pH 2.5, containing 1 g/L sodium dodecyl sulfate) was used as the mobile phase and the flow rate was 1.2 mL/min. The method was proved to be linear in the ranges of 1.03-206 mg/L and 5-200 mg/L for dextromethorphan hydrobromide and pseudoephedrine hydrochloride, respectively. The relative standard deviations of intra-assay (n = 7) and inter-assay (n = 5) were 1.0% and 1.5% for pseudoephedrine hydrochloride analysis and 1.8% and 2.2% for dextromethorphan hydrobromide analysis. The recoveries of pseudoephedrine hydrochloride and dextromethorphan hydrobromide were 95.7%-98.7% and 100.0%-101.8%, respectively. The method has been successfully applied to the simultaneous determination of pseudoephedrine hydrochloride and dextromethorphan hydrobromide in composite pseudoephedrine hydrochloride dry suspension. PMID- 15881375 TI - [Isolation and preparation of sesquiterpenols from the Japanese cedar, Cryptomeria japonica D. Don, by preparative reversed-phase high performance liquid chromatography and its characterization]. AB - A method for the isolation of sesquiterpenols from the Japanese Cedar, Cryptomeria japonica D. Don by preparative reversed-phase high performance liquid chromatography (RP-HPLC) was established. The preparation of the sesquiterpenols was carried out on a preparative liquid chromatograph with a YWC-Pack C18 column, and the mobile phase was isopropanol-methanol-hexane-water (50:35:10:5, v/v) at a flow rate of 5.0 mL/min. The two sesquiterpenols, (-)-cubebol and (+)-2,7(14),10 bisabolatrien-1-ol-4-one, were separated by liquid chromatography and identified by spectroscopic analyses (ultraviolet (UV), infrared (IR), nuclear magnetic resonance (NMR), electrospray ionization mass spectrometry (ESI-MS)). The purities of the two sesquiterpenols were 98.7% and 99.1%, respectively. PMID- 15881376 TI - [Determination of the contents of resveratrols and piceids in wines by high performance liquid chromatography with solid phase extraction]. AB - A method has been developed for the determination of the contents of cis- and trans-resveratrols and piceids in wines by high performance liquid chromatography (HPLC) with solid phase extraction (SPE). The techniques of SPE including the activation of the SPE column, elimination of interference and elution conditions of samples were confirmed. The analytes were separated by using a Hypersil C18 column (4.6 mm i. d. x 250 mm, 5 microm) and acetonitrile-water as mobile phase with linear gradient elution. A photodiode array detector monitored at the detection wavelengths of 288 nm and 306 nm for cis- and trans-isomers, respectively. The external standard calibration curves were used in quantitation. There were good linear correlations between the concentrations and peak areas of the four components with the correlation coefficients of 0.9973-0.9999. The average recoveries were 97.4%-98.6% with relative standard deviations of 2.1% 3.2%. The detection limits were 0.002-0.005 mg/L. The method of SPE-HPLC eliminated a great deal of interferences in wines effectively by optimizing extraction conditions. The four isomers were well separated and the analytical results were improved well and truly. The results for determining 15 market wines showed that as an important bioactive substance, the resveratrol content has close relationships with brewing methods, varieties of grape and producing areas. The results also showed that it is a convenient, rapid, sensitive and accurate method. It can be used to analyze resveratrols and piceids in other natural samples. PMID- 15881377 TI - [Analysis of chloramphenicol, thiamphenicol and florfenicol in chicken by high performance liquid chromatography with electrospray ionization mass spectrometry]. AB - The method for the analysis of three chloramphenicols including chloramphenicol, thiamphenicol and florfenicol in chickens was developed by high performance liquid chromatography combined with electrospray ionization mass spectrometry (HPLC-ESI-MS-MS). The mass spectrometer was operated in the negative ion mode using multiple reaction monitoring for qualitative and quantitative analysis of these compounds at the same time. Only 1 g of sample was needed, and no solid phase extraction (SPE) procedure was adopted. The advantages of the method are simple operation, less organic chemicals consumed, and shorter operation time. The limits of detection (LOD) were 0.010 microg/kg, and the limits of quantitation (LOQ) were 0.100 microg/kg. The linear plots were obtained between 0.050 and 1.00 microg/L. Overall recoveries were between 69.0% and 92.8% with relative standard deviations between 6.3% and 12.9%. PMID- 15881378 TI - [Preparative isolation and purification of cinnamoyl-C-glycoside chromone from aloe vera by high-speed countercurrent chromatography]. AB - Aloe chromone is a group of anti-inflammatory and anti-tyrosinase constituents found in aloe vera leaves. High-speed countercurrent chromatography (HSCCC) is reported for the preparative isolation and purification of a chromone from aloe vera. The crude extract was obtained by a series of pretreatment of aloe vera leaves and extracted from decolorizing active carbon with methanol. Then the extract was distributed between dichloromethane and water, and the organic part was then subjected to HSCCC for the isolation of chromone constituents. The chromone compounds with a high performance liquid chromatographic grade (>95%) was isolated through two step HSCCC separations by employing two solvent systems composed of chloroform-methanol-water and dichloromethane-methanol-water at volume ratios of 4/3/2 and 5/4/2, respectively. The chromone was finally identified as cinnamoyl-C-glycoside chromone by ultraviolet (UV), fast atom bombardment mass spectrometry (FAB-MS), nuclear magnetic resonance (1H NMR and 13C NMR). PMID- 15881379 TI - [Study of large-volume electrokinetic injection of capillary zone electrophoresis by using transient isotachophoresis]. AB - A sample pre-concentration method that combined field-amplified sample injection (FASI) induced by acetonitrile with transient isotachophoresis was applied to achieve effective concentration and the separation of cations in high-salt samples. Several important factors are discussed, such as buffer systems, terminating ions, column effective length and injection time of sample and terminating solution. A capillary column of 50 microm i. d. x 65 cm (effective length of 50 cm) was employed. Solutions of 400 mmol/L LiAc-HAc (pH 4.5) and 400 mmol/L beta-alanine-HAc (pH 4.5) were used as buffer solution and terminating electrolyte, respectively. The injection times of sample solution and terminating electrolyte were 270 and 90 s, respectively. Compared with conventional electrokinetic injection, the sensitivity of the proposed method was improved about 280-fold. The detection limits of propranolol and metoprolol achieved 2 x 10(-3) and 8 x 10(-3) mg/L, respectively. PMID- 15881380 TI - [Restriction enzyme pattern analysis of mycobacteria DNA by capillary electrophoresis with laser induced fluorescence detection]. AB - A new method for rapidly detecting restriction enzyme pattern of mycobacterium deoxyribonucleic acid (DNA) by capillary electrophoresis with laser induced fluorescence detection (CE-LIFD) was developed. Polymerase chain reaction was used to amplify a 439 bp fragment of 65,000 (Mr) heat shock protein gene (hsp65) of mycobacterium. After digesting the amplification products by BstE II and Hae III respectively, the patterns of enzyme cleavaged products were detected by both CE-LIFD and agarose gel electrophoresis (AGE). The experimental parameters of CE were optimized. The restriction enzyme patterns of mycobacterium DNA can be detected under the optimum electrophoresis conditions: a coated capillary column with the length of 50 cm and 100 microm i. d., electrophoresis buffer of 45 mmol/L TBE (trihydroxymethyl aminomethane (Tris)-boric acid ethylenediaminetetraacetic acid (EDTA)) and 11 kV running voltage. The restriction enzyme patterns for eight species of mycobacteria were studied. Relative standard deviations of the relative migration times of the DNA segments were less than 3.6%. Compared with AGE, CE is more outstanding in resolution and detection time, and it can be applied as a more effective means for DNA restriction enzyme pattern analysis. PMID- 15881381 TI - [Determination of five nerve agents in contaminated serum by gas chromatography]. PMID- 15881382 TI - [Detection of antioxidant capacities of anti-aging health foods by head-space gas chromatography]. PMID- 15881383 TI - [Determination of furoic acid and furfuryl alcohol by reversed-phase high performance liquid chromatography]. PMID- 15881384 TI - [Detection of catechin in edible lotus rhizome by high performance liquid chromatography]. PMID- 15881385 TI - [Analysis of active ingredients in Chinese hawthorn fruits by high performance liquid chromatographic method]. PMID- 15881386 TI - [Determination of the residue of quizalofop-p-tefuryl in soybeans by high performance liquid chromatography]. PMID- 15881387 TI - [Determination of furazolidone residue in aquatic products by high performance liquid chromatography]. PMID- 15881388 TI - [The network resources of ion chromatography]. PMID- 15881389 TI - [Further development (MDCK) of live cold-adapted influenza vaccine: cultivation of vaccine strains in production fermenters]. AB - Optimal conditions were developed for cultivating the cold-adapted reassortant live influenza vaccine (CARLIV) in MDCK cells, which were in their turn cultivated in fermenters with serum-free medium and microcarrier. The use of MDCK cells meets all national and WHO requirements to continuous cells used in the production of biological preparations. CARLIV cultivated under such conditions well preserve their ts-mutations and mutation, which entail substitutions of amino acids, in all CARLIV genome segments. Provided the cultivation conditions are optimal, the output of multivalent CARLIV in a 101 fermenter can reach 100000 doses. PMID- 15881390 TI - [Temporary acquisition of intra-chain disulphide bonds by nucleocapsid protein of influenza virus]. AB - The in vitro reducing agents were shown to promote the NP-NP association and to stabilize the NP oligomers, which dissociate when heated in non-reducing buffer. This confirms that non-covalent linkages in electrophoresis stabilize the influenza virus NP oligomers. The mobility of pulse-labeled and chased NPs in PAGE as well as their sensitivity to protease were investigated. The S-S bonds reduce at later stages of conformational maturation of NP; the disulfide containing NP transforms itself into an NP free of S-S bonds with non-covalently linked NP-oligomers being subsequently formed. Presumably, the early disulfide dependent stage in NP maturation is needed for the correct NP-NP association and for the protection of early monomeric NPs against protease action. PMID- 15881391 TI - [Genetic and phenotypic analysis of heterogeneous population of a cold-adapted donor of the A/Leningrad/134/17/57 (H2N2) attenuation and of the donor-based reassortant influenza vaccine strains]. AB - Cold-adapted (CA) temperature sensitive and attenuated virus A/Leningrad/134/17/57 (H2N2) (Len/17) has been recently used in Russia as a donor of internal genes in the preparation of reassortant vaccine strains of CA live influenza vaccine (LIV) for all age groups. The Len/17 population was found to be heterogeneous and to be made up of clones, which differ by combinations of mutations in internal genes. Around 50% of the Len/17 population had clones with all 8 coding mutations in internal genes. The others were made up of clones with mutation combinations, which were different from the original Len/17. The PCR restriction method was used to analyze 5 clones of Len/17 and 8 LIV vaccine strains. There were no Ala-86-Thr mutation in the M2 protein in 4 clones and 3 vaccine strains. The PB-1 gene of 4 clones and 3 vaccine strains had a mutation encoding Met-317-IIe more typical of a more attenuated virus A/Leningrad/134/47/57 (H2N2) (Len/47). The NP protein of a clone had a mutation Leu-341-IIe also typical of Len/47. However, neither the absence of mutation in the M2 gene nor an extra mutation in the PB1 gene affected the attenuation extent of reassortant CALIV. PMID- 15881392 TI - [Construction of DNA-fragments' libraries with complete genomes of different variola strains]. AB - Libraries of hybrid plasmids carrying DNA fragments of complete genomes of 8 variola virus strain from the Russian Collection belonging to 2 epidemical types and isolated in various geographic regions of the world were obtained. Genomic sequences of variola virus can be thus preserved for a long time in a biologically safe form and provide the research work on studying the genetic organization of this unique virus and on developing modern methods for rapid detection of variola virus and other orthopoxviruses. PMID- 15881393 TI - [Effects of cytokines on the immunogenic properties of hepatitis A vaccine]. AB - Cytokines were found, in experiments with guinea pigs, to have a stimulating action on the immunogenic potency of hepatitis A vaccine (Hep-A-in-Vac). The most pronounced effects were produced by rhIL-1b, rhTNF-alpha, thymosin-a1, the "neothym" hybrid protein and immunophan. Injections of cytokines as of adjuvants stimulated the formation of antibodies titers that exceeded 2-10-fold those observed in control animals immunized by Hep-A-in-Vac alone. Immunization of guinea pigs made alongside with injections of the above cytokines ensured a 100% seroconversion in animals after the administration of drugs was completed. The number of seropositive animals in the control group was 75-89%. PMID- 15881394 TI - [Detection of sapoviruses in children with acute gastroenteritis in Moscow in 2002-2003]. AB - Sapoviruses were found, for the first time, to be circulating in children with acute gastroenteritis in the city of Moscow. On the basis of a genetic analysis, they were classified as belonging to genotypes 1 and 2. Two groups of sapoviruses that are essentially different from the strains presented now at the GenBank NCBI were described within the case study. PMID- 15881395 TI - [Individual changes of gene expression in the interferon system in human blood cells due to amixin and cycloferon]. AB - The action of amixin and cycloferon on the expression of genes in the systems of interferon (IF) and cell apoptosis (CA) was studied by semi-quantitative RT-PCR in human blood microsamples before and after the administration of the drugs. Individual changes were determined in the transcription activity of genes of IF (alpha, beta, gamma), enzymes 2',5' oligoadenylatesynthetase (OAS), RNSase L, dsRNA-dependent proteinkinase (dsPK) and of CA effectors (FasAg, bcl-2, gamma actin) registered dynamically in 24 h and 48 h. The activity parameters of IF genes were compared with the results of biological titration of IF activity in blood samples in vivo and in vitro. A pronounced ability of cycloferon to stimulate selectively the activity of genes of human IF, type I (beta IF--by 100 times and alpha IF--by 10 times), without affecting essentially the activity of other genes in blood cells, was detected. Amixin was found to inhibit the titration of genes with high activity levels. (alpha-, beta-IF, RNAases L, bcl-2 and gamma-actin). The antiviral and IF-induced properties of the drug are explained to a great extent by the apoptotic effect (activation of genes Fas, gamma-IF, OAS and affected transcription of gene bcl-2). A positive correlation was observed between the processes of activation of IF-genes transcription and the production of the total circulating IF. Antagonistic relations between type I and II IFs in human blood cells were shown. PMID- 15881396 TI - [Morphological and immunohistochemical changes in tissues of the lung, myocardium and kidney in experimental West Nile fever]. AB - An experimental infection of mice with West Nile virus (WNV) showed pronounced dystrophic changes in tissues of the kidneys and myocardium as well as expression of WNV antigens in cells of the lungs, kidneys and myocardium, which can denote tropism of WNV to tissues of the lungs, kidneys and myocardium. PMID- 15881397 TI - [PCR-based detection of proviral DNA of bovine immunodeficiency virus]. AB - A set of primers was developed to detect by polymerase chain reaction (PCR) the proviral DNA of bovine immunodeficiency virus (BIV). A short fragment of 101 bp BIV gene was selected as a target for primers; sequences of proviral DNA isolated from both a cell culture with BIV and from lymphocytes of an experimentally infected animal were known for the fragment. An amplicon of an expected size was detected by standard PCR in a transformed cell series of bovine testicles with Florida 112 BIV DNA, and in a plasmid DNA with a cloned proviral DNA of R29 BIV. Described in the paper are the results of a theoretical comparison of primers used in the detection of BIV by PCR. The presence of non-complementary nucleotides in the set of "primer-single stranded amplicon" was shown to bring about false positive results in the detection of BIV by PCR. No 1500 bp PCR product was detected after PCR with a synthesized pair of primers and with 100% homology for all known BIV isolates complementary to env gene. Finally, the issue of how to detectVIR in clinical samples obtained from experimentally and naturally infected is discussed. PMID- 15881398 TI - [Development and certification of libraries of the MDCK continuous cell line for production of influenza vaccine]. AB - Seeding and working banks of the continuous MDCK cell culture suitable for the production of cultured influenza vaccine were created and deposited at liquid nitrogen temperature at the "Vector" State Research Center of Virology and Biotechnology. The MDCK cell culture was shown to have morphology typical of the discussed cell line; it does not have any alien agents and is oncogenically safe; its enzimogram and karyotype are typical of the donor line; finally, its biological properties are stable during a long period of cultivation and its sensitivity to influenza virus is high, therefore, it can be recommended for the production of influenza vaccine. The continuous MDCK cell line was certified at Tarasevich Committee and was recommended by the MIBP Committee, Russia's Health Ministry, for its use as a substrate in the production of diagnostic and preventive immunoglobulins. PMID- 15881399 TI - [Sensitivity of methods of titration of the vaccine strain of porcine fever virus]. AB - Methods of titration of the CS vaccine strain of classical swine fever virus were compared in vitro and vivo. The titration in the TL and PK-15 cell culture without cytopathic effect is based on the detection of virus antigen by labeled antibodies. The infection intensity in the cell culture virtually correlated with the antigenic and immunogenic activity of dry vaccine used for swine. PMID- 15881401 TI - Preparation of human recombinant kinesin heavy chain and epitope mapping of its structural domains. AB - The isolation of the cDNA sequence encoding the human neuronal kinesin (a force generating motor protein which transports various membrane organelles along microtubules in an ATP-dependent manner) heavy chain (nKHC) and the construction of expression vectors to produce the full-length nKHC and its domains in Escherichia coli is described. By tuning up the conditions for the expression of nKHC, a sufficient amount of the soluble protein intragenously tagged with 6xHis tag was obtained and purified by nickel chromatography. The recombinant structural domains of nKHC, including the motor domain (FKHC1--amino acids 1 330), the microtubule binding domain (FKHC2--amino acids 174-315) and the coiled coil stalk domain (FKHC3--amino acids 331-906) were used to determine the epitope location for monoclonal antibodies KN-01, KN-02, and IB II raised against different kinesin heavy chains. The antibodies were shown to recognize epitopes located in the stalk domain of nKHC and represent thus useful probes for this domain. PMID- 15881402 TI - Ten new temperate bacteriophages of Citrobacter youngae. AB - In a cross-test, we examined 55 strains of Citrobacter youngae against each other as potential producers of temperate bacteriophages and as potential sensitive indicators for them. Ten strains (18.2 %) showed the production of phages. Seven different strain-specific spectra of activity (from 1 to 11 strains each) were found. Phage production by 6 strains was inducible with mitomycin C, in 4 strains it was not inducible. The plaques of the phages were more or less turbid, without a lytic halo, tiny to small, 0.2-1.3 mm in diameter. Using a polyclonal, specific anti-lambda serum, all 10 phages were found to be clearly distinct from E. coli lambda phage, the phage 31/47 showing the highest neutralization titre of all. Interspecific tests with 15 strains of 8 species of Enterobacteriaceae revealed not a single case of activity of Citrobacter phages towards any of them. Five phage-immune clones lysogenized with 5 of the phages kept their remaining phage sensitivity spectra, though extended by sensitivity to 1-3 phages; 2 of these strains acquired also sensitivity to phage lambda. The phages belong to the morphotypes of Myoviridae (6 phages) and Siphoviridae (4 phages), with head diameters of 51-58 nm and tail length of 97-173 nm. Three strains produced corpuscular bacteriocins. PMID- 15881400 TI - DNA microarrays--techniques and applications in microbial systems. AB - Genome projects produce a huge amount of sequence information. As a result, the focus of genomics research is turning toward deduction of functional information about newly discovered genes. Thus structural genomics paves the way for a new discipline called functional genomics by providing the information required for microarray manufacture. Microarray technology is the result of automation and miniaturization in the detection of differential gene expression. By using this technology one can make a parallel analysis of RNA abundance and DNA homology for thousands of genes in a single experiment. Over the past several years, this unique technology has been used to explore hundreds transcriptional patterns and genome differences for a variety of microbial species. Applications of microarrays extend beyond the boundaries of basic biology into diagnostics, environmental monitoring, pharmacology, toxicology and biotechnology. We describe comprehensive nature of DNA microarray technology with emphasis on fabrication of DNA microarrays and application of this technology in biological environment with primary accent on microbial systems. PMID- 15881403 TI - Identification of a holin encoded by the Streptomyces aureofaciens phage micro1/6; functional analysis in Escherichia coli system. AB - An open reading frame encoding an 88 amino acid protein was present downstream of the previously characterized endolysin of Streptomyces aureofaciens phage micro1/6. Structural analysis of its sequence revealed features characteristic for holin. This open reading frame encoding the putative holin was amplified by polymerase chain reaction and cloned into the expression vector pET-21d(+). Synthesis of the holin-like protein resulted in bacterial cell death but not lysis. The holmicro1/6 gene was able to complement the defective lambda S allele in the nonsuppressing Escherichia coli HB101 strain to produce phage progeny, This fact suggests that the proteins encoded by both phage genes have analogous function, i.e. the streptomycete holin induces nonspecific lesions in the cytoplasmic membrane, through which the lambda endolysin gains an access to its substrate, the cell wall. The concomitant expression of both S. aureofaciens holmicro 1/6 and lambda endolysin in E. coli resulted in abrupt cell lysis. This result provided further evidence that the product of holmicro 1/6 gene is a holin. PMID- 15881404 TI - Optimization of a two-plasmid system for the identification of promoters recognized by RNA polymerase containing Mycobacterium tuberculosis stress response sigma factor, sigmaF. AB - The previously established two-plasmid system in Escherichia coli for the identification of Mycobacterium tuberculosis promoters that are recognized by RNA polymerase containing the stress response sigma factor sigmaF was optimized. Expression of the M. tuberculosis sigmaF encoded by sigF gene was under the control of the isopropyl beta-D-thiogalactopyranoside (IPTG)-dependent Ptrc promoter. A low level of IPTG induced a nontoxic but sufficient level of sigmaF to interact with the core enzyme of RNA polymerase. Such an RNA polymerase holoenzyme recognized the known sigmaF-dependent promoter, usfXp1, which was cloned in the compatible promoter probe plasmid, upstream of a promoterless lacZalpha reporter gene. Primer extension analysis of the usfXp1 promoter in the E. coli two-plasmid system after IPTG-induced expression of M. tuberculosis sigF revealed a transcription start point that was identical as in M. tuberculosis. This new system has been shown to be useful for identification of M. tuberculosis sigmaF-dependent promoters. PMID- 15881405 TI - Sample processing effect on polymerase chain reaction used for identification of Campylobacter jejuni. AB - Model samples of Campylobacter jejuni for polymerase chain reaction (PCR) were prepared by rapid and simple procedures consisting of centrifugation, proteinase K treatment, Chelex 100 treatment, and boiling lyses. A PCR based on specific amplification of the variable sequence of 16S rRNA gene was performed using Tth DNA polymerase and the PCR products were visualized by agarose gel electrophoresis. The assay allowed the detection of 10 CFU/mL C. jejuni in the physiological saline and 100 CFU/mL in the basic Park and Sanders broth. PMID- 15881406 TI - Parameiosis in Aspergillus nidulans in response to doxorubicin. AB - The recombinagenic effect of doxorubicin (an anticancer agent that impairs DNA synthesis and causes chromosome breaks) was used to induce parameiotic events in Aspergillus nidulans. Heterokaryons formed with master strains and uvs mutants were inoculated with and without doxorubicin. Haploid segregants (parameiotics and parents) and aneuploids were selected as heterokaryon-derived visible sectors. Among parameiotic segregants, recombinants by intergenic mitotic crossing-over and recombinants by chromosome-independent segregation were found. Whereas segregants of the former type were obtained only with doxorubicin, those of the latter type were recovered both with and without the drug. PMID- 15881407 TI - Physiological dendrogram of Claviceps spp. based on sucrose metabolism in submerged cultures and its comparison with phylogenetic tree. AB - Sixteen isolates of Claviceps spp. were analyzed for the production of polysaccharides, oligosaccharides, and sucrose metabolism under conditions of submerged fermentation. Physiological markers calculated by the Verhulst-Pearl law were used for hierarchical cluster analysis. Low correlation was found between physiologically based dendrogram and phylogenetic analysis constructed from an alignment of rDNA sequences. To confirm the intraspecific uniformity of physiological markers three isolates of C. africana from different hosts and locations were included. The influence of genotype, physiological variability, environmental location and habitat on metabolite production is discussed. PMID- 15881408 TI - Factors affecting killer activity of some yeast species occurring in Rokpol cheese. AB - Fourteen strains of Candida famata and 7 strains of C. sphaerica originating from blue-veined Rokpol cheese were studied for their ability to produce killer toxins against 3 strains of Yarrowia lipolytica selected as potential starter cultures for cheesemaking. All tested strains revealed killer activity in the presence of salt, at 14 degrees C and pH 4.6. Only one Y. lipolytica strain was resistant to C. famata and C. sphaerica killer toxins. PMID- 15881409 TI - Effect of biocides on S. cerevisiae: relationship between short-term membrane affliction and long-term cell killing. AB - The long-term action of recommended (RC) and near-recommended concentrations of several commercial biocides (Lonzabac 12.100, Genamin CS302D, benzalkonium chloride and 2-phenoxyethanol) on cells of S. cerevisiae wild-type strain DTXII was described using plating tests while short-term effects were determined using the potentiometric fluorescent probe diS-C3(3) that detects both changes in membrane potential and impairment of membrane integrity. A 2-d plating of cells exposed to 0.5xRC of benzalkonium chloride and Genamin CS302D for 15 min showed a complete long-term cell killing, with 2-phenoxyethanol the killing was complete only at 2xRC and Lonzabac caused complete killing at RC but not at 0.5xRC. The diS-C3(3) fluorescence assay performed immediately after a 10-min biocide exposure revealed several concentration-dependent modes of action: Lonzabac at 0.5xRC caused a mere depolarization, higher concentrations causing gradually increasing cell damage; benzalkonium chloride and Genamin CS302D rapidly damaged the membrane of some cells and depolarized the rest whereas 2-phenoxyethanol, which had the lowest effect in the plating test, produced a concentration dependent fraction of cells with impaired membranes. Cell staining slightly increased during the diS-C3(3) assay; addition of a protonophore showed that part of the remaining undamaged cells retained their membrane potential. Comparison of short-term and long-term data implies that membrane depolarization alone is not sufficient for complete long-term killing of yeast cells under the action of a biocide unless it is accompanied by perceptible impairment of membrane integrity. The results show that the diS-C3(3) fluorescence assay, which reflects the short term effects of a biocide on cell membranes, can be successfully used to assess the microbicidal efficiency of biocides. PMID- 15881410 TI - Characterization of esculin-positive Pseudomonas fluorescens strains isolated from an underground brook. AB - A group of sixteen esculin-positive fluorescent pseudomonads isolated from an underground brook flowing through a cave complex was characterized by biotyping, multiple enzyme restriction fragment length polymorphism analysis of 16S rDNA (MERFLP), ribotyping and whole-cell fatty-acid methyl-esters analysis (FAME). All strains were phenotypically close to Pseudomonas fluorescens, but they revealed high biochemical variability as well as some reactions atypical for P. fluorescens species. Because identification of pseudomonads by of biochemical testing is often unclear, further techniques were employed. Fingerprints obtained by MERFLP clearly showed that all strains represent P. fluorescens species. Ribotyping separated the strains analyzed into four groups corresponding almost completely (with the exception of one strain) to the clustering based on biochemical profiles. FAME analysis grouped all the strains into one cluster together with the P. putida (biotype A, B), P. chlororaphis and P. fluorescens biotype F representatives, but differentiated them from other FAME profiles of all pseudomonads included in the standard library TSBA 40 provided by MIDI, Inc. PMID- 15881411 TI - Biotechnological preparation of the elaiophylin. AB - Sporulation conditions (sporulation period and the sporulation broth) of the elaiophylin producer Streptomyces melanosporofaciens were optimized. The best culture sporulation was on agar broths with dextrin or hydrolyzed corn starch as carbon sources. PMID- 15881412 TI - Quantitative aspects of lipopolysaccharide and cytokine requirements to generate nitric oxide in macrophages from LPS-hyporesponsive (Lps(d)) C3H/HeJ mice. AB - Due to a gene defect (Lps(d)), C3H/HeJ mice are known to be hyporesponsive to the immunobiological potential of lipopolysaccharide (LPS). We studied dose requirements for LPS, IFN-gamma, and cytokines TNF-alpha and IL-10 to produce nitric oxide (NO) in peritoneal macrophages (Mphi) from these animals. In contrast to the Lps(n) C3H/HeN mice, high concentrations of LPS (up to 5 microg/mL) or IFN-gamma (up to 5 ng/mL) by themselves were unable to activate NO production in C3H/HeJ Mphi. The failure to produce NO could not be overcome by addition of L-arginine or tetrahydropterin. The high-output NO biosynthesis was dose-dependently stimulated by combined administration of varying concentrations of IFN-gamma (50-5000 pg/mL) and LPS (approximately 1 ng/mL) or to a lesser extent by IFN-gamma plus TNF-alpha or TNF-alpha/IL-10. Formation of NO in C3H/HeJ MCO triggered by high concentration of LPS (approximately 1 microg/mL) given together with IFN-gamma (0.2-5 ng/mL) reached the values typical for Lps(n) C3H/HeN mice. While Mphi from C3H/HeN mice secreted TNF-alpha, IL-10, and IL-10 upon contact with a low dose of LPS (1 ng/mL), C3H/HeJ Mphi required high concentration of LPS (5 microg/mL) to enhance the secretion of the cytokines. Yet, this dose remained ineffective to stimulate IFN-gamma in Mphi from C3H/HeJ mice. It can be presumed that one of the important factors influencing their deficient ability to form NO is a failure of Mphi to produce IFN-gamma upon LPS contact. PMID- 15881413 TI - Experimental colitis does not increase the prevalence of ANKENT, a spontaneous joint disease in mice. AB - A possible relationship between intestinal inflammation and joint disease development was investigated. Clinical symptoms of colitis--diarrhea and rectal bleeding--were confirmed by findings of inflammatory processes in the colon in dextran sodium sulfate-treated mice and joint ankylosing enthesopathy (ANKENT) developed in 12.8 % mice with chronic colitis and 13.6 % mice in the control group. Consequently no significant difference in ANKENT frequency was found between mice with and without chronic colitis and the occurrence of ANKENT in both groups was typical for conventional conditions. ANKENT cannot be triggered solely a generalized inflammatory process in the gut. PMID- 15881414 TI - Expression of inflammatory markers in pig amnion after intraamniotic infection with nonpathogenic or enteropathogenic Escherichia coli. AB - The pig amnion was in vivo intraamniotically infected with E. coli for 10 h at 80 85 d of gestation either with the nonpathogenic O86 strain or enteropathogenic O55 strain. TNF-alpha, IL-10, IL-1beta and IFN-gamma were determined in amniotic fluids by ELISA, the expression of cytokines and some other inflammatory markers was determined by immunohistochemistry. Intraamniotic infection induced high levels of TNF-alpha in amniotic fluids which correlated with bacterial virulence whereas IL-10 was induced only by O86. The IL-1beta level did not increase significantly and was expressed in all infected membranes. IFN-gamma was negligible or absent. TNF-alpha, IL-12p40, calprotectin, HSP65 and gp91phox were found by immunohistochemistry only in amnion membranes infected with the enteropathogenic strain 055. PMID- 15881415 TI - alpha5beta1 integrins and fibronectin are involved in adherence of the Pseudomonas aeruginosa ER97314 clinical strain to A549 cells. AB - The role of fibronectin (Fn) and its natural receptors alpha5beta1 integrins in the interaction of P. aeruginosa with A549 epithelial cells was compared in the clinical isolate ER97314 and the reference PAK strain. Both strains expressed functional type IV pili, as shown by the results of the twitching motility assay. The ER97314 strain was highly adherent to immobilized Fn (640 000+/-20 000 CFU per well) while the PAK strain adhered less efficiently (70 000+/-10 000 CFU per well). Both strains adhered to A549 cells (33 400+/-1200 and 1200+/-100 CFU per well, for PAK and ER97314, respectively), only the PAK strain being significantly internalized (9430+/-2020 CFU per well). Cytochalasin D and genistein significantly decreased bacterial adherence of the 2 strains and caused also a significant decrease in PAK internalization. This inhibitory activity was not related to changes in the expression of alpha5beta1 integrins. Antibodies to Fn and alpha5beta1 integrins inhibited the adherence of the ER97314 strain but had no significant effect on PAK interaction with human cells. These findings suggest that only some P. aeruginosa strains can target Fn and their natural receptors alpha5beta1 integrins for adherence to A549 cells. PMID- 15881416 TI - Antibiotic resistance and virulence factors among clinical and food enterococci isolated in Slovakia. AB - The resistance to antibiotics and the distribution of virulence factors in enterococci isolated from traditional Slovak sheep cheese bryndza was compared with strains from human infections. The occurrence of 4 enterococcal species was observed in 117 bryndza-cheese isolates. The majority of strains were identified as E. faecium (76 %) and E. faecalis (23 %). Several strains of E. durans and 1 strain of E. hirae were also present. More than 90 % of strains isolated from 109 clinical enterococci were E. faecalis, the rest belonged to E. faecium. The resistance to 6 antimicrobial substances (ampicillin, ciprofloxacin, higher concentration of gentamicin, nitrofurantoin, tetracycline and vancomycin) was tested in clinical and food enterococci. A higher level of resistance was found in clinical than in food strains and E. faecium had a higher resistance than E. faecalis; no resistance to vancomycin was detected. The occurrence of 3 virulence associated genes, cylA (coding for hemolysin), gelE (coding for gelatinase) and esp (coding for surface protein) was monitored. Differences were found in the distribution of cylA gene between clinical and bryndza-cheese E. faecalis strains; in contrast to clinical strains (45 %), cylA gene was detected in 22 % of food isolates. The distribution of 2 other virulence factors, gelE and esp, was not significantly different in the two groups of E. faecalis strains. cylA and gelE genes were not detected in E. faecium but more than 70 % of clinical E. faecium were positive for esp, even thought none of the 79 E. faecium cheese isolates contained this gene. PMID- 15881417 TI - Evaluation of metabolic control using a continuous subcutaneous glucose monitoring system in patients with type 1 diabetes mellitus who achieved insulin independence after islet cell transplantation. AB - This study evaluated the Medtronic MiniMed Continuous Glucose Monitoring System (CGMS) in patients with type 1 diabetes mellitus who underwent successful islet cell transplantation (ICT). The results are compared to standardized self monitoring (SMBG) of hyperglycemia and mean amplitude of glycemic excursions (MAGE). We studied 19 patients (mean age 40.0 +/- 6.7 years) in three groups: six patients post-ICT, seven patients awaiting ICT, and six normal volunteers (controls). Continuous glucose monitoring post-ICT showed remarkable glucose stability compared with patients awaiting ICT. The CGMS group showed modestly higher glucoses (mean 111.5 mg/dl) compared with controls (88 mg/dl). Postprandial glucoses in ICT recipients rarely exceeded 180 mg/dl and were similar to controls. There was no difference in asymptomatic hypoglycemia between control and post-ICT groups. However, a higher incidence of hypoglycemia was observed in patients awaiting ICT. HbA1c and MAGE pre- and post-ICT were 8.3 +/- 0.9% and 6 +/- 0.3% (p < 0.001) and 109 +/- 34 and 41 +/- 11 (p < 0.001), respectively. No complications were associated with CGMS. This study suggests ICT significantly improves metabolic control and rate of hypoglycemia when compared with controls and patients awaiting ICT. Similar improvement in metabolic control was observed with SMBG, HbA1c, and MAGE. Although CGMS was not demonstrated to be a superior tool for routine assessment in ICT, it is very helpful in special clinical situations. PMID- 15881418 TI - Prolonged allogeneic islet graft survival by protoporphyrins. AB - Transplantation of islets of Langerhans in patients with type 1 diabetes allows for improved metabolic control and insulin independence. The need for chronic immunosuppression limits this procedure to selected patients with brittle diabetes. Definition of therapeutic strategies allowing permanent engraftment without the need for chronic immunosuppression could overcome such limitations. We tested the effect of the use of protoporphyrins (CoPP and FePP), powerful inducers of the cytoprotective protein heme-oxygenase 1 (HO-1), on allogeneic islet graft survival. Chemically induced diabetic C57BL/6 mice received DBA/2 islets. Treatment consisted in peritransplant administration of CoPP or saline. Islets were either cultured in the presence of FePP or vehicle before implant. Short-course administration of CoPP led to long-term islet allograft survival in a sizable proportion of recipients. Long-term graft-bearing animals rejected third-party islets while accepting a second set donor-specific graft permanently, without additional treatment. Preconditioning of islets with FePP by itself led to improved graft survival in untreated recipients, and provided additional advantage in CoPP-treated recipients, resulting in an increased proportion of long-term surviving grafts. Preconditioning of the graft with protoporphyrins prior to implant resulted in reduction of class II expression. Administration of protoporphyrins to the recipients of allogeneic islets also resulted in transient powerful immunosuppression with reduced lymphocyte proliferative responses, increased proportion of regulatory cells (CD4+CD25+), decreased mononuclear cell infiltrating the graft, paralleled by a systemic upregulation of HO-1 expression. All these mechanisms may have contributed to the induction of donor-specific hyporesponsiveness in a proportion of the protoporphyrin-treated animals. PMID- 15881419 TI - Encapsulation of islets in rough surface, hydroxymethylated polysulfone capillaries stimulates VEGF release and promotes vascularization after transplantation. AB - The transplantation of encapsulated islets of Langerhans is one approach to treat type 1 diabetes without the need of lifelong immunosuppression. Capillaries have been used for macroencapsulation because they have a favorable surface-to-volume ratio and because they can be refilled. It is unclear at present whether the outer surface of such capillaries should be smooth to prevent, or rough to promote, cell adhesions. In this study we tested a new capillary made of modified polysulfone (MWCO: 50 kDa) with a rough, open-porous outer surface for islet transplantation. Compared with free-floating islets, encapsulation of freshly isolated rat islets affected neither the kinetics nor the efficiency of glucose induced insulin release in perifusion experiments. Free-floating islets maintained insulin secretion during cell culture but encapsulated islets gradually lost their glucose responsiveness and released VEGF. This indicated hypoxia in the capillary lumen. Transplantation of encapsulated rat islets into diabetic rats significantly reduced blood glucose concentrations from the first week of implantation. This hypoglycaemic effect persisted until explantation 4 weeks later. Transplantation of encapsulated porcine islets into diabetic rats reduced blood glucose concentrations depending on the islet purity. With semipurified islets a transient reduction of blood glucose concentrations was observed (2, 8, 18, 18 days) whereas with highly purified islets a sustained normoglycaemia was achieved (more than 28 days). Explanted capillaries containing rat islets were covered with blood vessels. Vascularization was also observed on capillaries containing porcine islets that were explanted from normoglycaemic rats. In contrast, on capillaries containing porcine islets that were explanted from hyperglycemic rats a fibrous capsule and lymphocyte accumulations were observed. No vascularization on the surface of transplanted capillaries was observed in the absence of islets. In conclusion, encapsulated islets can release VEGF, which appears to be an important signal for the vascularization of the capillary material. The rough, open-porous outer surface of the polysulfone capillary provides a site well suited for vascular tissue formation and may allow a prolonged islet function after transplantation. PMID- 15881420 TI - Human marrow stromal cells enhance connexin43 gap junction intercellular communication in cultured astrocytes. AB - Human marrow stromal cells (hMSCs) provide functional benefit in rats subjected to stroke. Astrocytes are coupled into a cellular network via gap junction channels, predominantly composed of connexin-43 (Cx43) proteins. Astrocytes are believed to play a vital role in neuroprotection by providing energy substrates to neurons and by regulating the concentrations of K+ and neurotransmitters via gap junctions. We therefore investigated the effect of factors secreted by hMSCs on gap junction intercellular communication (GJIC), expression of Cx43, and phosphorylation of Cx43 in an astrocyte cell culture system. Exposing rat cortical astrocytes to various concentrations of hMSC conditioned medium, we demonstrate that hMSCs produce soluble factors that significantly increase astrocytic GJIC, measured by the scrape-loading dye transfer method. Immunohistochemistry and Western blot showed increased Cx43 expression concomitant with altered GJIC. As the PI3K/Akt signaling pathway has been demonstrated to alter gap junction expression and GJIC, we selectively blocked phosphoinositide 3-kinase (PI3K). Addition of the PI3K inhibitor LY294002 decreased GJIC and Cx43 expression in astrocytes. These inhibitory effects of LY294002 were countered by the addition of hMSC conditioned media. Furthermore, coculturing hMSCs with rat astrocytes increased astrocyte GJIC in a manner dependent upon the hMSC/astrocyte ratio. These findings demonstrate that hMSCs secrete soluble factors that increase GJIC of astrocytes through upregulation of Cx43, and indicate a mechanistic role for PI3K. PMID- 15881421 TI - Dopaminergic reinnervation of the globus pallidus by fetal nigral grafts in the rodent model of Parkinson's disease. AB - The current neural transplantation strategy for Parkinson's disease (PD) involves the dopaminergic reinnervation of the striatum (STR). Although up to 85% reinnervation of the STR has been attained by neural transplantation, functional recovery in animal models and transplanted patients is incomplete. This limitation may be due to an incomplete restoration of the dopaminergic input to other basal ganglia structures such as the external segment of the globus pallidus (GPe, homologue of the rodent GP), which normally receives dopaminergic input from the substantia nigra (SN). As part of our investigation into a multiple grafting strategy for PD, we have explored the effects of dopaminergic grafts in the GP of rodents with unilateral 6-hydroxydopamine (6-OHDA) lesions. In this experiment, lesioned rats received either 300,000 fetal ventral mesencephalic (FVM) cells or a sham injection into the GP. Functional assessment consisted of rotational behavior at 3 and 6 weeks posttransplantation. A fluorogold tracer study was conducted to rule out any behavioral improvement due to striatal outgrowth of the GP graft. Sections were stained for glial fibrillary acidic protein (GFAP) to assess the degree of trauma in the GP by the graft in comparison to the sham injection. Immunohistochemistry for tyrosine hydroxylase (TH) was performed after transplantation to assess graft survival. Animals with GP grafts demonstrated a significant improvement in rotational behavior at 3 and 6 weeks posttransplantation (p < 0.05) while sham control animals did not improve. All animals receiving FVM cells showed TH-immunoreactive grafts in the GP posttransplantation. TH-positive neurons in the GP showed no double labeling with an intrastriatal injection of fluorogold, indicating that behavioral improvement was not due to striatal innervation by the GP graft. These observations suggest that functional recovery was the result of dopaminergic reinnervation of the GP and that this nucleus may be a potential target for neural transplantation in clinical PD. PMID- 15881422 TI - Inhibition of chromatin condensation prevents transgene silencing in a neural progenitor cell line transplanted to the rat brain. AB - The use of ex vivo gene therapy in the central nervous system has so far suffered from transgene downregulation. Condensation of the transgenic sequences has been proposed to be a mechanism involved in this silencing. In this study we inhibited either histone deacetylation or DNA methylation in neural progenitor cell lines, transduced with a lentiviral vector carrying green fluorescent protein (GFP), prior to grafting them into the rat striatum. The expression of GFP was significantly higher in grafts pretreated with either of the inhibitors. After 1 week in vivo we detected an 11-fold increase in the number of GFP-expressing cells due to the inhibition of DNA methylation in vitro with azadeoxycytidine and a ninefold increase when inhibiting histone deacetylation with trichostatin A. This suggests that a pretreatment paradigm could be used to increase efficacy of ex vivo delivery of a therapeutic protein locally in the brain. PMID- 15881423 TI - In vivo noninvasive monitoring of a tissue engineered construct using 1H NMR spectroscopy. AB - Direct, noninvasive monitoring of tissue engineered substitutes containing live, functional cells would provide valuable information on dynamic changes that occur postimplantation. Such changes include remodeling both within the construct and at the interface of the implant with the surrounding host tissue, and may result in changes in the number of viable cells in the construct. This study investigated the use of 1H NMR spectroscopy in noninvasively monitoring the viable cell number within a tissue engineered construct in vivo. The construct consisted of mouse betaTC3 insulinomas in a disk-shaped agarose gel, surrounded by a cell-free agarose gel layer. Localized 1H NMR spectra were acquired from within implanted constructs, and the total choline resonance was measured. Critical issues that had to be addressed in accurately quantifying total choline from the implanted cells included avoiding signal from host tissue and correcting for interfering signal from diffusing solutes. In vivo NMR measurements were correlated with MTT assays and NMR measurements performed in vitro on explanted constructs. Total choline measurements accurately and noninvasively quantified viable betaTC3 cell numbers in vivo, in the range of 1 x 10(6) to more than 14 x 10(6) cells, and monitored changes in viable cell number that occurred in the same construct over time. This is the first study using NMR techniques to monitor viable cell numbers in an implanted tissue substitute. It established architectural characteristics that a construct should have to be amenable to NMR monitoring, and it set the foundation for future in vivo investigations with other tissue engineered implants. PMID- 15881424 TI - Isolated hepatocyte transplantation for Crigler-Najjar syndrome type 1. AB - Crigler-Najjar syndrome type 1 (CN1) is an inherited disorder characterized by the absence of hepatic uridine diphosphoglucuronate glucuronosyltransferase (UDPGT), the enzyme responsible for the conjugation and excretion of bilirubin. We performed allogenic hepatocyte transplantation (AHT) in a child with CN1, aiming to improve bilirubin glucuronidation in this condition. A 9-year-old boy with CN1 was prepared with plasmapheresis and immunosuppression with prednisolone and tacrolimus. When a graft was made available, 7.5 x 10(9) hepatocytes were isolated and infused into the portal vein percutaneously. After 2 weeks phenobarbitone was added to promote the enzymatic activity of UDPGT of the transplanted hepatocytes. Nocturnal phototherapy was continued throughout the studied period. Total bilirubin was considered a reliable marker of allogenic cell function. There was no significant variation of vital signs nor complications during the infusion. Mean +/- SD bilirubin level was 530 +/- 38 micromol/L before and 359 +/- 46 micromol/L after AHT (t-test, p < 0.001). However, the introduction of phenobarbitone was followed by a drop of tacrolimus level with increase of alanine aminotransferase (ALT) and increase of bilirubin. After standard treatment of cellular rejection bilirubin fell again but from then on it was maintained at a greater level. After discharge the patient experienced a further increase of bilirubin that returned to predischarge levels after readmission to the hospital. This was interpreted as poor compliance with phototherapy. Only partial correction of clinical jaundice and the poor tolerability to nocturnal phototherapy led the parents to refuse further hepatocyte infusions and request an orthotopic liver transplant. After 24 months the child is well, with good liver function on tacrolimus and prednisolone-based immunosuppression. Isolated AHT, though effective and safe, is not sufficient to correct CN1. Maintenance of adequate immunosuppression and family compliance are the main factors hampering the success of this procedure. PMID- 15881425 TI - Effect of microcapsule composition and short-term immunosuppression on intraportal biocompatibility. AB - With higher nutrient and oxygen supply and close contact to blood, the portal vein is a possible alternative to the peritoneal cavity for transplantation of encapsulated cells. Data regarding intraportal biocompatibility of microcapsules are lacking. Microcapsules were built from five alginate types differing in their molar mass and mannuronic/guluronic acid ratios by complex formation with divalent cations (barium or calcium) or mixtures of divalent cations and polycations. They were injected in the portal vein of rats, and cellular and fibrotic pericapsular infiltration thickness was measured 3 and 7 days after implantation. Overgrowth was characterized using various stainings or immunohistochemistry (hematoxylin and eosin, Giemsa, ED-1 for monocyte/macrophage, alpha-actin for myofibroblasts, CD31 for endothelial cells). The impact of short-term immunosuppression (gadolinium-chloride IV 20 mg/kg/day on days--1 and 4 as well as 10 days of rapamycin PO 1 mg/kg/day, tacrolimus PO 3 mg/kg/day, or combinations of rapamycin/tacrolimus or gadolinium/tacrolimus) was further assessed 3, 7, and 42 days after implantation. Overall, overgrowth increased from day 3 to day 7 (p < 0.05). Three and 7 days after implantation, polycation-containing microcapsules induced more reaction than microbeads (p < 0.0001 and p < 0.01). Considering polycation-free beads, barium-alginate induced the weakest reaction. Biocompatibility of microbeads was independent of mannuronic/guluronic acid ratio and molar mass of the alginate. Infiltration was mainly a monocyte/macrophage-rich foreign body reaction, but an eosinophil containing immunoallergic reaction was also observed. Short-term immunosuppression significantly reduced infiltration in all conditions and up to 42 days after implantation. Biocompatibility after intraportal infusion was best for barium-alginate microbeads and poorest for polycation-containing microcapsules. Short- and long-term overgrowth could be significantly reduced by short-term immunosuppression. PMID- 15881426 TI - Islet ontogeny determines glucose responsiveness. PMID- 15881427 TI - [Substantiated diagnostic properties of two-component analysis of rheogram]. AB - The paper presents a conclusive basis applicable to the parameters of two component analysis of biosignals of the pulse filling of the rheogram type conditioning the parameters' specific use. A new original rheographic parameter is suggested for the examined zone of vascular filling--local fraction of cardiac output. A verbal model of general hemodynamic pulse filling developed by the author previously and specified within the present cases study has been used for the purpose. The model was constructed as a specific description of the working cardiovascular system during cardiac cycles, which is based on the two-phase cardiac function, with respect to the active heart work both in the systolic and diastolic phases of the cardiac cycle. PMID- 15881428 TI - [Hemodialysis with biological object]. AB - The essence of the method of biodialysis (hemodialysis with biological object) developed and suggested by the authors for clinical use consists in that the healthy organism exerts, through a system of mass transfer, a therapeutic action on the sick organism. Blood from the affected and healthy organisms is perfused through individual mass exchangers (dialyzers, hemodiafilters and hemofilters), which are hydraulically connected by a circulating transport medium. Metabolites that accumulate in blood of the affected organism diffuse into the transport medium and, from there, into blood of the healthy organism, which metabolizes them. The reverse process occurs simultaneously: substances, whose concentration in blood of the sick organism is less versus the healthy organism, diffuse from blood of the healthy organism to blood of patient. The method suggested by us can be used in clinical practice for normalizing a variety of parameters in patients with hepatic and renal insufficiency. Besides, a number of substances can be transferred from the healthy donor to patient in the process of biodialysis, which opens promising potentialities for the treatment of many diseases. PMID- 15881429 TI - [Substantiated need in information support to therapeutic-and-diagnostic processes]. AB - On the basis of analysis it is proven as necessary to use information support in treatment and diagnosis at different-status hospitals to enhance the quality and efficiency of treatment. The used theoretical-and-empirical approach is based on major parameters of three therapeutic-and-diagnostic processes studied within different coordinate systems as well as on evaluation of total knowledge needed by practitioner and on complexity of formal treatment tasks. PMID- 15881430 TI - [A study of a two-energy bar of detectors for x-ray osteodensitometry]. AB - Described are the results of mathematic modeling for different types of detectors (with different scintillators for high- and low-energy spectrum ranges) as well as the results of experimental research. It was also analyzed of how a manufacturing technology of ZnSe(Te) and CsI(Tl)-based scintillators affects the detector-bar resolution ability. Finally, the influence of signals in canals of detectors on the signals in neighboring canals was investigated. PMID- 15881431 TI - [An integrated approach to the occupational specialization of handicapped teenagers]. AB - Presented in the paper are issues connected with occupational specialization of handicapped children and with psychophysiological diagnosis including the program "Yong disabled should have equal opportunities" developed at the Center for rehabilitation of handicapped children, Arzamas, for those who are restricted in their choice of occupation due to health. PMID- 15881432 TI - [Biofeedback of human heart rate: adaptive module]. AB - Ecological issues, that are not in line with the adaptive opportunities of man, prompt the designing of computer-based systems for psychophysiological support of traditional methods of prevention, diagnostics and rehabilitation. An adaptive module of biofeedback was developed for monitoring the human heart rate. The system can be applied in a variety of medical spheres, like evaluation of adaptive potentialities of different body systems, enhancing of the body's reserve and of its efficiency; mobilization of latent or lost abilities to adapt; training of skills of relaxation, self-regulation and self-control; correction of the condition of persons at different stages of occupational, ecological psychological and social stress; and preliminary psychophysiological preparation for responsible operator tasks. PMID- 15881433 TI - [Examination of spectral parameters of heart rate in junior schoolchildren at mental work]. AB - Described in the paper are the results of a study of spectral parameters of variable heart rate (VHR) in 67 junior schoolchildren, aged 8-11, obtained alongside with the Landolt test. It was reliably demonstrated that the total capacity of the VHR spectrum and of spectral indices was decreased at the time of test; however, it went up subsequently to the initial level or topped it. The results of investigation of spectograms of VHR recorded in the course of the Landolt non-verbal test explain individual features related with the heart rate monitoring and with adaptive potentialities of schoolchildren. PMID- 15881434 TI - [Features of microfocus radiography in medical diagnostics]. AB - The paper describes the features of image capture with microfocus X-ray technologies in medical diagnostics, including the main reason of decreasing radiation doses received by different bone structures of patients. PMID- 15881435 TI - [Technical specificity of synchronous registration of pulse]. AB - Analyzed in the paper is mutual influence of channels in synchronous registration of pulse signals from human radial arteries made by a device whose transducers share a common base. Such influence is conditioned by the base mobility, which in its turn is caused by the fact that the device rests on soft tissues of the hand and that the fixation elements work by cross rigidity. Transducers must be fixed separately and the impact of their initial force on signal registration zones needs to be eliminated by design-based separation of zones of registration and fixation to enable the use of radiophysical methods in signal processing. PMID- 15881436 TI - [Differential sensitivity of vision to flicker rate]. AB - A method is suggested for measurement of the differential sensitivity of vision to the light flicker rate. The best accuracy of the method with subsequent presentation of the increment and decrement flicker rates was experimentally established. PMID- 15881437 TI - [A survey of medical equipment displayed at the "Healthcare-2004" exhibition]. AB - Surveyed in the paper is medical equipment displayed, November 29-December 3, 2004, in the Exhibition Hall in Krasnaya Presnya within the framework of the "Healthcare, Medical Equipment and Drugs-2004" Exhibition. The survey is made for the below categories of medical equipment: instruments and apparatuses; X-ray, neonatal, ophthalmologic and stomatological equipment; medical instruments; laboratory medical equipment; equipment for rehabilitation of patients and disabled and mobile medical equipment. A conclusion is made on that a lot of novel high-quality and high-tech medical equipment has emerged in the Russian market. The diversified equipment displayed at the Exhibition enables any medical facility to choose for its needs the most appropriate medical equipment, instruments and spares. PMID- 15881438 TI - [Vertical stability of schoolchildren with spastic diplegia before and after complex physical rehabilitation]. AB - The paper presents the results of a study of vertical stability of 18 children and teenagers (mean age 12.5) with spastic diplegia as observed before and after complex treatment including the use of the method of phase electrostimulation of muscles and of methods developed at the Physical Exercise Department, Pediatric Neurological Hospital No. 18. The patients were examined by Romberg's test on the KSK-123 stabilographic unit with open and closed eyes. The results testify indirectly to a treatment-conditioned improvement in the CNS and receptor apparatus as observed during regulation of the vertical posture balance in the examinees. Thus, with visual monitoring, the mean square deviation in the frontal plane dropped on the average by 48% and, without such monitoring, it went down by 34%; in the sagittal plane, the values decreased by 51% and 35%, respectively. The area of statokynesogram shrank by 38% with open eyes and by 27% with closed eyes. The posttreatment prevalence of sagittal fluctuations in the body's center of gravity as observed versus frontal fluctuations is also typical of the health condition of the examinees. PMID- 15881439 TI - [Power-consumption for storage of donor plasma]. AB - Costs of electric power used for storage of freshly frozen plasma (FFP) were compared for refrigerators most widely used in Russian medical facilities. The minimal costs (2763.43 R) are those obtained for storage of 5000 FFP units in 12 plasma freezers, MDF-U536 (Sanyo, Japan). When 30,000 plasma units are stored the cost efficiency of power consumption is higher for the TELEDOOR freezer (Germany) versus MDF-U536. It can be assumed that, as the electric power cost increases, it will be more attractive to store FFP in liquid nitrogen vapor (MABAG, Germany). The above technology is registered in Russia; however, its application is still problematic due to economic reasons. PMID- 15881440 TI - [The future belongs to the union of medical enterprises and associations]. PMID- 15881441 TI - Clinical negligence and the legal services commission. PMID- 15881442 TI - Presidential address. Medical evidence in the criminal courts. PMID- 15881443 TI - Justice and incompetence. PMID- 15881444 TI - Return to work following whiplash and back injury: a review and evaluation. AB - The purpose of this study is to review the reported return-to-work rates following whiplash and back-injury. The return-to-work rates for the 71 relevant studies that were reviewed varied from 29% to 100% with a median of 67%. The results suggest considerable residual return-to-work potential for persons with whiplash and back injury. Return-to-work rates were substantially higher for motor vehicle (96%) compared to work-related (71%) studies and also considerably higher for whiplash (95%) compared to back injuries (65%). It is suggested that these indices may form potential benchmarks for personal injury claims outcomes. The purpose of this paper is to analyse the published rates of return to work following whiplash and back injury. This criterion of return to work is not without limitations or controversy and Pfingsten, Hildebrandt Leibing, Franz and Saur (1997) noted: "... a high variability exists in data on return-to-work. Results often range from either no return to a very high return rate, with an approximate average of 68% following multimodal treatment procedures...". PMID- 15881445 TI - Who knows best? PMID- 15881446 TI - Since when is being Jewish irrelevant? PMID- 15881447 TI - Practice management. PMID- 15881449 TI - Where are all the Jewish dental students? PMID- 15881448 TI - Partners in creation. PMID- 15881450 TI - Mothers' awareness about accidents among toddlers. AB - A study to identify the knowledge of mothers on prevention of accidents among the toddlers in a selected urban community of Mangalore was conducted in the year of 1999 at Dr. M. V. Shetty Institute of Health Sciences, Vidyanagar, Mangalore, affiliated to Rajiv Gandhi University of Health Sciences, Bangalore. PMID- 15881451 TI - Dengue fever: diagnosis, prevention & control. PMID- 15881452 TI - Ionic liquid for high temperature headspace liquid-phase microextraction of chlorinated anilines in environmental water samples. AB - Based on the non-volatility of room temperature ionic liquids (IL), 1-butyl-3 methylimidazolium hexafluorophosphate ([C4MIM][PF6]) IL was employed as an advantageous extraction solvent for high temperature headspace liquid-phase microextraction (LPME) of chloroanilines in environmental water samples. At high temperature of 90 degrees C, 4-chloroaniline, 2-chloroaniline, 3,4 dichloroaniline, and 2,4-dichloroaniline were extracted into a 10 microl drop of [C4MIM][PF6] suspended on the needle of a high-performance liquid chromatography (HPLC) microsyringe held at the headspace of the samples. Then, the IL was injected directly into the HPLC system for determination. Parameters related to LPME were optimized, and high selectivity and low detection limits of the four chlorinated anilines were obtained because the extraction was performed at high temperature in headspace mode and the very high affinity between IL and chlorinated anilines. The proposed procedure was applied for the analysis of the real samples including tap water, river water and wastewater samples from a petrochemical plant and a printworks, and only 3,4-dichloroaniline was detected in the printworks wastewater at 88.2 microg l(-1) level. The recoveries for the four chlorinated anilines in the four samples were all in the range of 81.9-99.6% at 25 microg l(-1) spiked level. PMID- 15881453 TI - Highly porous solid-phase microextraction fiber coating based on poly(ethylene glycol)-modified ormosils synthesized by sol-gel technology. AB - The preparation and characteristics of solid-phase microextraction (SPME) fibers coated with Carbowax 20M ormosil (organically modified silica) are described here. Raw fused silica fibers were coated with Carbowax 20M-modified silica using sol-gel process. Scanning electron micrographs of fibers revealed a highly porous, sponge-like coating with an average thickness of (8 +/- 1) microm. The sol-gel Carbowax fibers were compared to commercial fibers coated with 100 microm polydimethylsiloxane (PDMS) and 65 microm Carbowax-divinylbenzene (DVB). Shorter equilibrium times were possible with the sol-gel Carbowax fiber: for headspace extraction of the test analytes, they ranged from less than 3 min for benzene to 15 min for o-xylene. Extraction efficiencies of the sol-gel Carbowax fiber were superior to those of conventional fibers: for o-xylene, the extracted masses were 230 and 540% of that obtained with 100 microm PDMS and 65 microm Carbowax-DVB fibers, respectively. PMID- 15881454 TI - Equilibrium in-fibre standardisation technique for solid-phase microextraction. AB - This note describes a fundamental investigation into solid-phase microextraction (SPME) using a standard loaded into the fibre coating as a means of internal standardisation for the analysis of samples contained in vials. The loading of reproducible amounts of standards into a non-porous SPME fiber was investigated. It was found that spiking low milligram quantities of standards such as benzene, toluene, ethylbenzene, xylenes (BTEX) and/or naphthalene into a few grams of pump oil sealed in a 20 mL vial provided an excellent standard generator. A single solution allowed over a hundred standard loadings with a reproducibility of <4% R.S.D. When a fiber, loaded with the standard(s) was introduced into a sample vial, extraction of analytes into the fiber and desorption of the standard(s) into the sample matrix occur simultaneously. Quantification was then based on the equilibrium distribution of the standards and the analytes between the fibre coating and the sample matrix in the vial. A comparison of equilibration profiles obtained using traditional internal standardisation and the in-fibre approach generally showed the same equilibration behaviour. The developed method was successfully used to correct for matrix effects in the BTEX analysis of a wine sample. PMID- 15881455 TI - New approach of solid-phase microextraction improving the extraction yield of butyl and phenyltin compounds by combining the effects of pressure and type of agitation. AB - A new methodology for the simultaneous and fast solid-phase microextraction (SPME) of butyl- and phenyltin compounds, as ethylated derivates, is proposed in this paper. The effects of pressure and type of agitation during headspace SPME sampling are evaluated and discussed on the basis of thermodynamic considerations. Quantitative structure-activity relationships were used to estimate analytes partition coefficients allowing to explain the different behaviours experimentally observed. SPME sampling conditions including mechanical stirring and reduced pressure result in simultaneous higher efficiency (detection limits especially lowered for phenyltins up to a eight-fold reduction) and shorter sampling time (two-fold reduction). PMID- 15881456 TI - Liquid-phase microextraction based on carrier mediated transport combined with liquid chromatography-mass spectrometry. New concept for the determination of polar drugs in a single drop of human plasma. AB - Recently, we demonstrated for the first time liquid-phase microextraction (LPME) of polar drugs based on carrier mediated transport. In this new extraction technique, selected analytes were extracted as ion-pairs from small volumes of biological samples, through a thin layer of a water immiscible organic solvent immobilised in the pores of a porous hollow fibre (liquid membrane), and into a microl volume of an acidic aqueous acceptor solution placed inside the lumen of the hollow fibre. In the current paper, this new extraction technique was combined with liquid chromatography-mass spectrometry (LC-MS) for the first time. Carrier mediated LPME was evaluated for several new model drugs (0.01 or = 65 years). STUDY DESIGN: Open, 12-month, follow-up study of 444 patients, treated with indapamide SR, who were responders and/or achieved target BP levels following a 3-month, randomised, controlled, double-blind short-term comparison of indapamide SR versus hydrochlorothiazide 25 mg and amlodipine 5 mg. RESULTS: The long-term decrease in systolic blood pressure (SBP)/diastolic blood pressure (DBP) after 12 months follow-up with indapamide SR was -24.0/-13.1 mmHg from baseline (M0). The percentage of patients that achieved target BP levels (DBP < 95 mmHg, SBP < or = 160 mmHg) was 80.1% [84.3% for isolated systolic hypertension (ISH) subgroup], and the response rate (BP < 140/90 mmHg or decrease in supine diastolic BP > or = 10 mmHg or in supine systolic BP > or = 20 mmHg) 81.5%. Blood pressure (BP) remained stable throughout the 12 months follow-up period (M3-M15), whatever the previous treatment received during the 3-month, doubleblind period (M0-M3). Clinical and biological acceptability was good. A low occurrence of withdrawals (7.2%), was reported. CONCLUSION: Over the course of the long-term, 12-month follow-up study, indapamide SR was shown to be an effective and well tolerated antihypertensive therapy, even after a switch from amlodipine or hydrochlorothiazide, in patients aged 65 years-80 years with systolo-diastolic hypertension (SDH) or ISH. PMID- 15881475 TI - Assessing the cost-effectiveness of COX-2 specific inhibitors for arthritis in the Veterans Health Administration. AB - OBJECTIVE: This study was designed to assess the cost-effectiveness of cyclooxygenase-2 specific (COX-2) inhibitors (rofecoxib and celecoxib) over nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) in high-risk arthritis patients from the perspective of the Veterans Health Administration (VA). METHODS: This literature-based economic analysis (with data summarized from MEDLINE-indexed and other published sources, FDA reports, and data on file at VA San Diego Healthcare System) compared rofecoxib and celecoxib to NSAIDS in two arthritis patient populations considered at higher risk of developing clinically significant upper gastrointestinal events (CSUGIEs): (1) patients of any age with previous medical history of perforation/ulcer/bleed (PUB); and (2) patients 65 years and older (regardless of history of PUB). Two outcome measures were reported: (1) incremental cost per CSUGIE averted over 1 year; and (2) incremental cost per quality-adjusted life-year (QALY) gained, considering both the mortality and morbidity associated with gastrointestinal (including CSUGIEs) and cardiovascular-related adverse events. When possible, costs were modeled to reflect the VA perspective. Sensitivity analyses were conducted to test the robustness of the analysis. RESULTS: Compared to NSAIDS, rofecoxib and celecoxib increased costs but reduced the incidence of CSUGIE. Cost per CSUGIE avoided were $7476 and $16,379 (in patients with a PUB history) and $14,294 and $18,376 (in patients aged > or = 65 years) for celecoxib and rofecoxib, respectively. In both populations, celecoxib was associated with a cost per QALY less than $50,000. In contrast, rofecoxib was found to cost more and result in a net QALY loss, due in particular to the increase in the risk of cardiovascular complications, and was therefore considered cost-ineffective. Results were most dependent on assumptions about the incidence of cardiovascular events and CSUGIE and the COX-2 inhibitors' acquisition price. CONCLUSIONS: This analysis suggests that COX-2 inhibitors may be cost-effective from the perspective of the VA. However, cost-effectiveness appears to depend less on the specific characteristics of the high-risk target population considered but more on the agent evaluated. Celecoxib appears to be an alternative to traditional NSAIDs in the patient populations studied. PMID- 15881476 TI - Once daily clarithromycin extended-release vs twice-daily amoxicillin/clavulanate in patients with acute bacterial sinusitis: a randomized, investigator-blinded study. AB - OBJECTIVE: To compare the efficacy and tolerability of clarithromycin extended release (ER) to amoxicillin/ clavulanate in patients diagnosed with acute bacterial sinusitis. RESEARCH DESIGN AND METHODS: In a controlled, multicenter, investigator-blinded study, 437 ambulatory patients at least 12 years old with signs/symptoms and radiographic findings of acute sinusitis were randomized to receive clarithromycin ER 1000 mg once daily or amoxicillin/ clavulanate 875 mg/l25 mg twice daily for 14 days. MAIN OUTCOME MEASURES: Clinical and bacteriological response rates were determined at a test-of-cure visit, which was conducted up to 10 days following the completion of treatment. Radiological response was assessed at a follow-up visit. RESULTS: The clinical cure rate in clinically evaluable patients was 98% (184/188) in the clarithromycin ER group and 97% (179/185) in the amoxicillin/clavulanate group (95% CI for the difference in rates [-2.4%, 4.7%]). Clinical cure was sustained at the follow-up visit (96% for each treatment group). The pathogen eradication rates were 94% (61/65) in the clarithromycin ER group and 98% (61/62) in the amoxicillin/clavulanate group (95% CI for difference in rates [-12.0%, 2.9%]). The radiological success rate was 94% (172/183) in both the clarithromycin ER and amoxicillin/clavulanate groups (95% CI for difference in rates [-4.9%, 4.9%]). Symptomatic improvement or relief was observed as early as 2 days-5 days after the initiation of study drug, with a statistically significantly higher resolution rate of sinus pressure (p = 0.027) and improvement/resolution rate of nasal congestion (p = 0.035) during treatment with clarithromycin ER. The resolution/improvement rate at the test-of-cure visit for each treatment group was > or = 94% for the primary acute sinusitis signs/symptoms, with a statistically significantly higher resolution/improvement rate of purulent nasal discharge with clarithromycin ER (p = 0.010). Both study drugs had a positive and rapid impact on quality of life. Patients reported a high level of satisfaction and probability of using either study antibiotic again, and health care resource use was low, with slightly fewer sinusitis related physician and outpatient visits required by patients in the clarithromycin ER group (p = 0.055). The treatment groups were comparable with respect to incidence of drug-related adverse events. CONCLUSION: In this multinational population of patients with acute bacterial sinusitis, clarithromycin ER was comparable, and for selected measures superior, to amoxicillin/clavulanate based on clinical, bacteriological, and radiological responses as well as quality of life measures, satisfaction with antibiotic therapy, and health care resource utilization. PMID- 15881477 TI - Recent developments in the management of overactive bladder: focus on the efficacy and tolerability of once daily solifenacin succinate 5 mg. AB - BACKGROUND: Overactive bladder (OAB) is a highly prevalent symptom complex that may be extremely distressing to the patient, and can be associated with co morbidities and reduced quality of life (QoL). One of the major pathophysiological causes of OAB is overactivity of the detrusor muscle, mediated via muscarinic receptors in the bladder. Urgency is the defining symptom of OAB, yet a significant proportion of patients also suffer from incontinence, which is the most distressing symptom to the patient. As such, restoration of continence should be a primary treatment goal. However, effective treatments should also impact on the other key symptoms of OAB, such as micturition frequency and urgency. Non-pharmacologic interventions to treat OAB can be effective but require patients to be highly motivated. In terms of pharmacologic therapy, treatment with an antimuscarinic agent is the mainstay of current therapy. Solifenacin succinate is a once-daily oral antimuscarinic for the treatment of OAB. The recommended dose is 5mg once daily and can be increased to 10 mg once daily if 5 mg is well tolerated. OBJECTIVES: This paper reviews clinical experience with solifenacin 5 mg in patients with OAB as this is the recommended dose according to FDA product labeling. FINDINGS: In Phase 3 studies, based on data captured in 3-day micturition diaries, greater than half of patients who were incontinent at baseline no longer reported experiencing incontinence episodes after 12 weeks of double-blind treatment with solifenacin 5 mg. Furthermore, compared with placebo, solifenacin treatment resulted in statistically significant reductions in incontinence episodes, micturition frequency and urgency episodes, with significant increases in volume voided (based on an analysis of key symptom outcomes in two pooled Phase 3 studies presented here). The most common treatment-related adverse events were expected anticholinergic side effects (dry mouth, constipation, and blurred vision), and these were generally mild to moderate. Discontinuation rates due to adverse events in the treatment and placebo groups were comparable. CONCLUSION: Solifenacin 5 mg was found to be efficacious and had an acceptable tolerability profile in patients with OAB in these trials and this treatment may provide QoL benefits to patients. PMID- 15881478 TI - Cost effectiveness of using carboxymethylcellulose dressing compared with gauze in the management of exuding venous leg ulcers in Germany and the USA. AB - OBJECTIVE: To assess the cost effectiveness of using carboxymethylcellulose dressing (CMCD; Aquacel Hydrofiber) compared to gauze in the management of exuding venous leg ulcers in Germany and the USA. DESIGN AND SETTING: This was a modelling study performed from the perspective of payers (i.e. the sickness funds in Germany and the community sector in the USA). METHODS: Clinical outcomes attributable to managing exuding venous leg ulcers were obtained from the published literature in the English language. These data were combined with resource utilisation estimates derived from a panel of clinicians enabling us to construct two decision models depicting the management of venous leg ulcers with CMCD or gauze over 18 weeks in Germany and the USA. The models were used to estimate the cost effectiveness of CMCD compared to gauze in the management of exuding venous leg ulcers in both countries. MAIN OUTCOME MEASURES AND RESULTS: Starting treatment with CMCD instead of gauze in both Germany and the USA is expected to heal 30% of ulcers within 18 weeks compared to 13% with gauze (p = 0.003). The healthcare cost of starting treatment with CMCD or gauze in Germany is expected to be Euro2020 and Euro 2654 respectively at 18 weeks. Additionally, the healthcare cost of starting treatment with CMCD or gauze in the USA is expected to be $3797 and $5288 respectively at 18 weeks. Hence, using CMCD instead of gauze is expected to increase the probability of healing within 18 weeks by 130% and reduce healthcare costs by at least 24%. The healthcare cost of managing CMCD-treated patients was less than that of gauze-treated patients in both countries due to decreased nursing and physician costs associated with a lower frequency of CMCD dressing changes compared to gauze dressing changes. If it were assumed that treatment with gauze in both countries heals 30% of ulcers within 18 weeks (i.e. is identical to CMCD), then the expected healthcare cost of using gauze would be reduced by only 3% (from Euro2654 to Euro2562 in Germany and from $5288 to $5148 in the USA). CONCLUSION: Within the limitations of our model, starting management of an exuding venous leg ulcer with CMCD instead of gauze is the cost effective strategy in both Germany and the USA. Moreover, the purchase price of a leg ulcer dressing should not be used as an indication of the cost effectiveness of a given method of care. PMID- 15881479 TI - Ocular penetration of moxifloxacin 0.5% and gatifloxacin 0.3% ophthalmic solutions into the aqueous humor following topical administration prior to routine cataract surgery. AB - A prospective, double-masked, randomized, parallel-group study (n = 25) was conducted to examine the ocular penetration of moxifloxacin 0.5% ophthalmic solution and gatifloxacin 0.3% solution into the aqueous humor following topical administration prior to routine cataract surgery. One drop of antibiotic was instilled every 10 min for four doses beginning 1 h prior to surgery. Preliminary results showed aqueous humor concentrations for moxifloxacin that were significantly greater (p < 0.01) than those for gatifloxacin. PMID- 15881480 TI - The mechanism of action of omega-3 fatty acids in secondary prevention post myocardial infarction. AB - BACKGROUND: Omega-3 fatty acids from fish and fish oils can protect against coronary heart disease (CHD), which is still the most common cause of death in the Western economies. Evidence from epidemiological and case cohort studies indicate that consumpton of fatty fish and omega-3 fatty acids reduces the risk of cardiovascular mortality. OBJECTIVE: This article briefly reviews the evidence regarding omega-3 fatty acids and CHD and outlines the mechanisms through which omega-3 fatty acids might confer cardiac benefits over and above the standard secondary prevention strategies. CONCLUSION: The conclusion reached is that omega 3 fatty acids play a significant role in secondary prevention post-myocardial infarction. The mechanisms through which two of these omega-3 fatty acids, eicosapentaenoic acid and docosahexanoic acid, exert their action appear to be distinct and adjuvant to the available standard secondary prevention therapies. The role to be played by the administration of a newly licensed 90% concentrate EPA + DHA formulation (1 g/day capsule: Omacor) is explored. PMID- 15881481 TI - Modelling the long term cost effectiveness of clopidogrel for the secondary prevention of occlusive vascular events in the UK. AB - OBJECTIVE: To assess the long term cost effectiveness of clopidogrel monotherapy compared with acetylsalicylic acid (aspirin; ASA) monotherapy in patients at risk of secondary occlusive vascular events (OVEs) in the UK. DESIGN: Cost utility analysis based on clinical data from CAPRIE (a multicentre randomised controlled trial, involving 19185 patients); long-term effects were extrapolated beyond the trial period using a Markov model populated with data from UK observational studies. Health economic evaluation carried out from the perspective of the UK National Health Service. PARTICIPANTS: A representative cohort of 1000 UK patients aged 60 years (approximate mean age of the CAPRIE population), with the qualifying diagnoses of myocardial infarction, ischaemic stroke and peripheral arterial disease, who are at risk of secondary OVEs (non-fatal myocardial infarction, non-fatal stroke or vascular death). INTERVENTIONS: Patients were assumed to receive treatment with either clopidogrel (75 mg/day) for 2 years followed by ASA (325 mg/day, average) for their remaining lifetime, or ASA alone (325 mg/day, average) for life. MAIN OUTCOME MEASURES: Incremental cost per life year gained and incremental cost per quality-adjusted life year (QALY) gained. RESULTS: In the base case, the incremental cost effectiveness of clopidogrel versus ASA in this population is estimated at 18888 pounds per life year gained and 21 489 pounds per QALY gained. Multiple deterministic and probabilistic sensitivity analyses suggest the model is robust to variations in a wide range of input parameters. CONCLUSION: Two years of treatment with clopidogrel can be considered a cost effective intervention in patients at risk of secondary OVEs in the UK. PMID- 15881482 TI - Comparison of rilmenidine and lisinopril on ambulatory blood pressure and plasma lipid and glucose levels in hypertensive women with metabolic syndrome. AB - OBJECTIVE: In previous studies, the I1 imidazoline specific agonist rilmenidine effectively lowered office blood pressure (BP) in patients with metabolic syndrome, improved glucose metabolism and did not demonstrate unfavourable effects on plasma lipids. The aim of the present study was to investigate the effects of 12weeks therapy with rilmenidine compared with the ACE inhibitor lisinopril on ambulatory BP, plasma lipid and fasting glucose levels in women with metabolic syndrome. RESEARCH DESIGN: Prospective randomised open-label, blinded end-points study. METHODS: Female patients (n = 51) with hypertension and other components of metabolic syndrome were treated with 1 mg rilmenidine (n = 24) or 10 mg lisinopril (n = 27), once- or twice-daily. Anthropometric measurements, office BP and heart rate (HR) measurements, ambulatory BP monitoring, lipid and fasting glucose assessment were performed before and after 12weeks of treatment MAIN OUTCOME MEASURES: Changes in ambulatory BP and HR, including 24-h, daytime and night-time values, and in lipids and glucose levels. All changes were adjusted for baseline values using the analysis of covariance method. RESULTS: Ambulatory 24-h systolic BP and diastolic BP were decreased significantly in the rilmenidine group (-11.9 +/- 1.9 and -7.7 +/- 0.8 mm Hg, p < 0.001) respectively and the lisinopril group (-11.0 +/- 1.8 and -6.7 +/- 0.7 mm Hg respectively, p < 0.001). There were no significant differences between the two groups. Rilmenidine reduced 24-h ambulatory HR (-3.6 +/- 0.8 bpm versus 0.3 +/- 0.8 bpm with lisinopril; p = 0.002). The reductions of day-time and night time BP were also significant for both treatment groups, but the rilmenidine group demonstrated a greater decrease in night-time diastolic BP (p = 0.046). Rilmenidine significantly increased HDL cholesterol and decreased fasting glucose levels (p = 0.009 and p = 0.012, respectively). HDL cholesterol tended to increase and fasting glucose tended to decrease in the lisinopril group. However, differences between groups were not significant. CONCLUSION: Rilmenidine has similar effects on ambulatory BP patterns in hypertensive women with metabolic syndrome as lisinopril. Rilmenidine compared with lisinopril significantly reduces ambulatory HR. In this study, rilmenidine and lisinopril demonstrate similar effects on plasma lipid and fasting glucose levels. PMID- 15881483 TI - Bioequivalence of liquid and freeze-dried recombinant human follicle-stimulating hormone. AB - OBJECTIVE: The bioequivalence and tolerability of freeze-dried and liquid formulations of recombinant human follicle-stimulating hormone (r-hFSH) filled-by mass were assessed in a crossover, open-label, randomised, single-centre, phase I bioequivalence study. METHODS: Following pituitary down-regulation with the gonadotrophin-releasing hormone agonist goserelin, healthy adult volunteers (18years-45years of age) received single subcutaneous injections of r-hFSH , 300 IU, from freeze-dried and liquid formulations in random order, separated by a 7 day washout period. Blood was obtained over 144 h for pharmacokinetic analysis. MAIN OUTCOME MEASURES: These were peak serum FSH concentrations (Cmax,), time to peak concentration (Tmax) and area under the concentration-time curve from zero to the last measurable concentration (AUCJ), local and systemic tolerability. RESULTS: Of 44 volunteers who underwent down-regulation, 39 (18 men, 21 women) completed the study. Cmax and AUClast were similar with the freeze-dried (mean 9.51 IU/L and 844 IU.h/L, respectively) and liquid (mean 8.99 IU/L and 841 IUh/L, respectively) formulations, whereas T was significantly higher with the liquid formulation (median 12h vs 15h, p = 0.0183). The 90% confidence intervals for the ratio of the treatment means for Cnw and AUC,=, were within the pre-defined bioequivalence range of 0.8-1.25. CONCLUSION: Both formulations were well tolerated with regard to both systemic and local adverse events. The freeze-dried and liquid formulations of r-hFSH are bioequivalent and show no significant differences in tolerability. Thus, the liquid formulation is expected to provide comparable efficacy and tolerability to the freeze-dried formulation in clinical use. PMID- 15881484 TI - Safety and efficacy of bedtime versus daytime administration of the miconazole nitrate 1200 mg vaginal ovule insert to treat vulvovaginal candidiasis. AB - OBJECTIVE: Most vaginal products are self-administered at bedtime. Safety, efficacy, and ' therapeutic cure with bedtime versus daytime treatment of vulvovaginal candidiasis (VVC) were assessed to potentially improve compliance and convenience with vaginal antifungal treatment. METHODS: A non-inferiority study evaluated 573 women with symptoms of WC, randomized to daytime or bedtime self-administration of single dose of miconazole nitrate 1200 mg vaginal ovule (Monistat-1). RESULTS: Subjects (n = 149) were evaluated for efficacy in the Daytime group and 163 in the Bedtime group. In the Daytime group, 86 (57.7%) participants achieved therapeutic cure, with 105 (70.5%) and 111 (74.5%) exhibiting mycologic and clinical cures, respectively. In the Bedtime group, 83 (50.9%) subjects achieved therapeutic cure, with 104 (63.8%) and 120 (73.6%) exhibiting mycologic and clinical cures, respectively (NS). There was no significant difference in the incidence of side effects between the two groups. Median estimated time to initial relief of itching, burning and irritation was similar for both. Miconazole nitrate 2% cream was also applied twice daily to the vulvar area for external symptoms for up to 7 days. Test-of-cure was assessed 21 days-30 days following administration of intra-vaginal drug. Time to vulvovaginal symptomatic relief and the effect of activity following daytime administration on therapeutic cure were also assessed. CONCLUSION: The efficacy against a broad spectrum of pathogens and acceptability of daytime and bedtime administration of miconazole nitrate 1200 mg vaginal ovule were similar. These findings offer women a convenient option for treatment for WC, whether self-managed or managed by their healthcare provider. PMID- 15881485 TI - Year-by-year analysis of cardiovascular events in the Multiple Outcomes of Raloxifene Evaluation (MORE) trial. AB - OBJECTIVE: To assess the effect of raloxifene 60 mg/day (RLX) on year-by-year cardiovascular (CV) events in postmenopausal women participating in the Multiple Outcomes of Raloxifene Evaluation (MORE) trial, a double-blind, placebo controlled osteoporosis treatment trial. RESEARCH DESIGN AND METHODS: Post hoc analysis, using data from participants receiving placebo (N = 2576) or RLX 60 mg/day (N = 2557) in MORE, was performed to determine the relative risk (RR, 95% CI) of CV events in each individual trial year. Analyses were performed for the overall cohort and for women in high and low risk subsets. Women were retrospectively assessed as high CV risk using established criteria and the remaining women were considered low CV risk. RESULTS: The incidence of CV events did not differ between the RLX and placebo groups in the overall cohort (RR 0.86, 95% Cl 0.64-1.15), or the low CV risk subset (RR 1.01, 95% Cl 0.70-1.46). In the high-risk subset, the incidence of CV events was less in the RLX group (RR 0.60, 95% Cl 0.38-0.95). There was no significant increase in CV risk during any single year in the RLX group for either the overall cohort or the low or high CV risk subsets. CONCLUSION: In this post hoc analysis, the risk of CV events was not increased in any single year of MORE in women taking RLX, either in the overall cohort or in the low and high CV risk subsets. PMID- 15881486 TI - The analgesic effect of etoricoxib relative to that of cetaminophen analgesics: a randomized, controlled single-dose study in acute dental impaction pain. AB - BACKGROUND: To compare the analgesic effect of single doses of etoricoxib 120 mg, oxycodone/ acetaminophen 10 mg/650 mg and codeine/ acetaminophen 60 mg/600 mg in acute pain using the dental impaction model. METHODS: In this randomized, double blind, placebo-controlled, parallel-group study, patients reported pain intensity and pain relief (16 times) and global scores (twice) during a 24-h period. The primary endpoint was the overall analgesic effect, total pain relief over 6 h (TOPAR6). Other endpoints were patient global evaluation, time to onset (2 stopwatch method), duration of analgesic effect (median time to and amount of rescue medication use). Tolerability was evaluated by overall and opioid-related (nausea and vomiting) adverse experiences. RESULTS: 302 patients (mean age 23; 63% women; 63 % White) were randomized to etoricoxib 120 mg, oxycodone/acetaminophen 10 mg/650 mg, codeine/acetaminophen 60 mg/600 mg, and placebo (2:2:1:1). Etoricoxib demonstrated significantly greater overall analgesic efficacy (TOPAR6) (13.2 units) versus oxycodone/acetaminophen (10.2 units); and codeine/acetaminophen (6.0 units); p < 0.001 for all. All active treatments were superior to placebo. Median time to onset was significantly (p < 0.001) shorter for oxycodone/acetaminophen (20 min) and numerically but not significantly shorter (p = 0.259) for codeine/acetaminophen (26 min) compared with etoricoxib (40 min). Etoricoxib (24 h) had a significantly longer lasting analgesic effect than oxycodone/acetaminophen (5.3 h), codeine/acetaminophen (2.7 h), and placebo (1.7 h) (p < 0.001 for all). Etoricoxib patients experienced fewer clinical adverse experiences than patients on oxycodone/acetaminophen and codeine/acetaminophen, specifically, significantly (p < 0.05) fewer episodes of nausea. CONCLUSION: Etoricoxib 120 mg provided superior overall analgesic effect with a smaller percentage of patients experiencing nausea versus both oxycodone/acetaminophen 10 mg/650 mg and codeine/acetaminophen 60 mg/600 mg. PMID- 15881487 TI - Lumiracoxib in the treatment of osteoarthritis, rheumatoid arthritis and acute postoperative dental pain: results of three dose-response studies. AB - OBJECTIVES: Overview of three dose-response studies demonstrating the efficacy of lumiracoxib, a novel COX-2 selective inhibitor, for chronic pain associated with osteoarthritis (0A), or rheumatoid arthritis (RA) and acute pain following dental extraction. METHODS: OA and RA: 4-week, randomized, placebo- and active controlled studies of similar design. Patients (OA, n = 583; RA, n = 571) received lumiracoxib 50 mg, 100 mg or 200 mg twice daily (bid), lumiracoxib 400 mg once daily (od), diclofenac 75 mg bid or placebo. Dental: 12-h, single-center, randomized, placebo- and active-controlled study. Patients (n = 202) received single oral doses of lumiracoxib 100 mg or 400 mg, ibuprofen 400 mg or placebo. MAIN OUTCOME MEASURES: OA: pain intensity (PI) in the target joint (visual analogue scale [VAS]) and WOMAC score at Week 4; RA: overall PI (VAS) and ACR20 response at Week 4; Dental: difference (PID, categorical and VAS) score over 12h post dose, time to onset of analgesia. RESULTS: Throughout the OA study, all lumiracoxib doses provided superior reductions in PI versus placebo and at Week 4, all lumiracoxib doses provided efficacy similar to each other and to diclofenac. In the RA study, lumiracoxib 100 mg bid, 200 mg bid and 400mg od were significantly better than placebo in PI at Weeks 1 and 2 (all p < 0.05) but demonstrated borderline significance at Week 4 (lumiracoxib 400 mg od, p = 0.06). In pain following dental surgery, PID scores for both lumiracoxib doses were superior to placebo from 1.5 h onwards and always comparable, or superior, to ibuprofen. Lumiracoxib 400 mg had the fastest onset of analgesia, measured as median time to confirmed first perceptible pain relief using the two-stopwatch method (37.4 min, superiority versus placebo, p < 0.001). Lumiracoxib was well tolerated in all studies. CONCLUSIONS: These studies provide initial evidence that lumiracoxib is an effective, well-tolerated agent for the treatment of chronic and acute pain. PMID- 15881488 TI - Effect of pioglitazone on metabolic syndrome risk factors: results of double blind, multicenter, randomized clinical trials. AB - OBJECTIVE: To evaluate the effect of pioglitazone, metformin, and/or sulfonylurea on metabolic syndrome and its component parameters after a year of treatment. METHODS: Adult patients with poorly controlled type 2 diabetes were enrolled in 4 multicenter, double-blind studies and received pioglitazone, sulfonylurea, metformin, or a combination of any 2 agents. Post hoc analyses were performed on data from patients with evaluations at baseline and week 52, and treatment groups were compared to determine change from baseline in metabolic syndrome status and its component variables by using the McNemar test and analysis of covariance, respectively. RESULTS: Most patients (72.1%) had metabolic syndrome at baseline. Change in the proportion of patients with metabolic syndrome status was significant in each monotherapy and the pioglitazone plus metformin combination groups. Pioglitazone alone or in combination with metformin resulted in a significantly greater percent decrease from baseline in triglycerides (pioglitazone vs. metformin, 10.3%; pioglitazone vs. sulfonylurea, 6.5%; pioglitazone plus metformin vs. sulfonylurea plus metformin, 13.4%; P < 0.05) and a greater percent increase from baseline in high-density lipoprotein (HDL) cholesterol (pioglitazone vs. metformin, 9.1%; pioglitazone vs. sulfonylurea, 12.6%; pioglitazone plus metformin vs. sulfonylurea plus metformin, 17.8%; P < 0.001) at week 52 than did the respective comparison groups. A significant decrease from baseline in the ratio of urinary albumin to creatinine was found only with pioglitazone monotherapy (-1.764 mg/ mmol; P < 0.001), which was significantly greater than the change in the metformin monotherapy group (2.1%; P < 0.05). Significant decrease in blood pressure was observed in the pioglitazone monotherapy and pioglitazone plus sulfonylurea groups, with no significant treatment group differences. CONCLUSIONS: Treatment with pioglitazone as monotherapy or combination therapy led to sustained, positive effects on important components of metabolic syndrome in patients with type 2 diabetes, independent of effects on blood glucose control and, as such, could be translated to potential for reducing the risk of cardiovascular disease. PMID- 15881489 TI - Share this issue of Nursing Economic$ with your HR and IT departments. PMID- 15881490 TI - Six-part series on the state of the RN workforce in the United States. AB - This series on the state of the nursing workforce focuses on describing the results of the national surveys of RNs. The series begins with an analysis of RNs' perceptions of the current nursing shortage and whether they perceive the shortage has gotten better or worse over the past few years. Future topics will include RNs' perceptions of being a nurse and how they perceive certain aspects of their jobs; RNs' awareness of the J&J Campaign and whether they think it has been effective; RNs' perceptions of the impact of the shortage on the quality of patient care in hospitals; and various issues related to the age and diversity of the nursing profession. PMID- 15881491 TI - Is the shortage of hospital registered nurses getting better or worse? Findings from two recent national surveys of RNs. AB - The results of two recent national surveys of RNs that were conducted at a time when the nursing shortage in the U.S. was in full force (2002) and 2 years later (2004) were compared. The findings provide a mixed assessment: on the one hand, there is evidence that the shortage has eased since 2002 and that there have been notable improvements in the lives of nurses; on the other hand, the shortage has had a negative impact on hospitals and nurses, and longstanding problems associated with the workplace environment remain. PMID- 15881492 TI - Nurse-physician communication: an organizational accountability. AB - Dysfunctional nurse-physician communication has been linked to medication errors, patient injuries, and patient deaths. The organization is accountable for providing a context that supports effective nurse-physician communication. Organizational strategies to create such a context are synthesized from the structural, human resource, political, and cultural frameworks of organizational behavior. PMID- 15881493 TI - The impact of hospice programs on U.S. hospitals. AB - This study demonstrates that hospice programs are more likely in communities with favorable economic factors and higher Medicare populations. Large hospitals with high occupancy rates and more clinical services use the hospice as an alternative to inpatient care thereby improving hospital efficiency and profitability. PMID- 15881494 TI - Analyzing fluctuating unit census for timely staffing intervention. AB - A computer information system, which can provide cost-saving measures useful in adequately allocating nursing staff in the face of decreasing patient stays, increasing patient admissions and discharges, and the unexpected transfers surrounding busy nursing units, is explored. PMID- 15881495 TI - Work excitement in nursing: an examination of the relationship between work excitement and burnout. AB - The results of this study found a significant relationship between burnout and the Work Excitement Model. This suggests that utilization of the Work Excitement Model by health care organizations may reduce nursing burnout and improve productivity and quality of care. PMID- 15881496 TI - As we lie in the bed we made: the malpractice and regulatory consequences of failing leadership. PMID- 15881497 TI - Expertise isn't the only answer to leadership in safety. AB - In the words of Covey (2004), we all have the power to decide to live a great life rather than a life of mediocrity. Unfortunately, in patient safety, we in health care have chosen to live a life of mediocrity because we have not made a commitment to error-free versus an error-tolerant industry. Compared to what other industries have accomplished in safety, we must make a commitment to do better. We must stop benchmarking ourselves against the median or average, which is mediocrity at its finest, and accept the challenge that an error-free environment is possible. In the words of Collins (2001), "Good is the enemy of Great" (p. 1). With new eyes that represent diverse cognitive skills, we will overcome our blindness to defects and move from good to great. PMID- 15881498 TI - The future for peer review. Florida's constitutional amendment chills quality community. AB - A new amendment to its state constitution has opened up Florida's peer review meeting notes to the public, with chilling effects on the entire peer review process. PMID- 15881499 TI - What skills does your board need? AB - Growing numbers of health care boards are evaluating their current and potential trustees to ensure that they have enough independence and the right skill mix to govern in a tougher environment. PMID- 15881500 TI - Trustee workbook 2. The balanced scorecard: a tool for maximizing board performance. PMID- 15881501 TI - Avoiding 'builder's remorse'. Maximizing the value of your facilities investments. AB - To make sure big health care facility investments don't end up as big capital mistakes, building projects must be linked to long-term organizational goals. PMID- 15881502 TI - George Winn: showing up and speaking up. Interview by Laurie Larson. PMID- 15881503 TI - The importance and paucity of trust in today's health care system. PMID- 15881504 TI - What do boards need to know? PMID- 15881505 TI - An introduction to dentists with special interests. AB - DwSIs are an NHS initiative. The Department of Health will be providing guidance to PCTs on how to assess a practitioner's suitability to provide specific services for the local healthcare community based on local need and circumstance. PMID- 15881506 TI - Trends in indirect dentistry: introduction. PMID- 15881507 TI - Trends in indirect dentistry: 1. Case selection. AB - Indirect restorations can provide a conservative, functional and aesthetic method for managing a patient's dentition. These restorations, however, are only as good as the environment in which they are placed. Many restorations, though technically satisfactory, may fail because assessment and subsequent treatment planning has been unsatisfactory. This article presents a philosophy of case selection mindful of factors related to the patient, operator and oral environment. The wrong treatment, however skilfully executed, may invite failure. PMID- 15881508 TI - Bacteria in the mouth. AB - There are many infectious agents found within the oral cavity. Bacteria form an important group of micro-organisms found in both healthy and diseased mouths. The current article will discuss those bacteria resident within the mouth and review the oral diseases with which they are associated. PMID- 15881509 TI - Why and how to get patients to stop smoking. AB - Despite the fact that smoking cigarettes harms oral and systemic health, there is little current evidence of successful primary dental care involvement in smoking cessation. This paper aims to present the arguments for developing the role of primary dental care in smoking cessation and reviews the current literature on how this can be achieved in practice. It is argued that it is timely to incorporate this rapidly developing evidence base on what works in smoking cessation into primary dental care activity. PMID- 15881510 TI - Impacted premolars. AB - Tooth impaction is not uncommon among the population. Its management is important aesthetically and functionally to the patient. Treatment ranges from regular review, extraction of the primary tooth, surgical exposure of the impacted tooth, with or without orthodontic traction, auto-transplantation to its surgical removal. Any decision taken should be judged on individual merit. PMID- 15881511 TI - The twin block appliance for the correction of Class II malocclusion. AB - A number of treatment approaches are available for the management of Class II malocclusion. Functional appliances are commonly used for the management of mandibular retrognathia in growing subjects. The twin block appliance is the most popular functional appliance in use within the United Kingdom. The aim of this article is to review the clinical management of the twin block appliance and to discuss some of the hard and soft tissue changes that accompany treatment with this appliance. PMID- 15881512 TI - Making a good impression: (a 'how to' paper on dental alginate). AB - Approximately 20,000 impressions are made with alginate annually at the University Dental Hospital of Manchester alone, and countless more wherever clinical dentistry is practised. A proportion of these are considered unsatisfactory for clinical use. The reason for these 'failures' is often poor operator technique, rather than an inherent problem of the material. With improved handling of the material and better clinical technique, failures can be avoided. This paper provides an overview of the use of alginate impression material in clinical dentistry. PMID- 15881513 TI - Physical signs for the general dental practitioner. PMID- 15881514 TI - Pulmonary vein imaging: comparison of 3D magnetic resonance angiography with 2D cine MRI for characterizing anatomy and size. AB - Pulmonary vein imaging is integral for planning atrial fibrillation ablation procedures. We tested the feasibility of quantifying pulmonary vein ostial diameter using two-dimensional cine cardiac magnetic resonance (2D cine CMR) and three-dimensional magnetic resonance angiography (3D MRA). Nine patients with a history of atrial fibrillation and 20 normal volunteers underwent 2D cine CMR and contrast-enhanced 3D MRA of pulmonary veins on a 1.5 T scanner. Pulmonary vein ostial diameters were measured and pulmonary vein vessel border sharpness was graded qualitatively. Both techniques provided excellent pulmonary vein imaging; however, 3D MRA was faster to perform. The average difference between the systolic and diastolic pulmonary vein diameter was 2.5 mm (23.2%, p < 0.0001) in normal volunteers and 2.2 mm (16.9%, p < 0.0001) in atrial fibrillation patients. The ostial diameter measurements by 3D MRA were significantly larger than on 2D cine CMR. Additionally, the pulmonary vein borders appeared sharper with 2D cine CMR compared to 3D MRA. In conclusion, the 2D images can resolve differences in diameter across the cardiac cycle, while the 3D images provide high quality anatomical depiction but blur borders due to pulsatile motion. We suggest a protocol combining 2D cine CMR and 3D MRA for comprehensive evaluation of pulmonary veins. PMID- 15881515 TI - Blood flow quantification in adults by phase-contrast MRI combined with SENSE--a validation study. AB - PURPOSE: To evaluate the feasibility of rapid free-breathing phase-contrast MRI (PC-MRI) at different in-plane resolutions combined with sensitivity encoding (SENSE) for flow quantification in the great arteries in healthy adult volunteers. METHODS: In 13 volunteers (mean age 33.0 +/- 7.4 years), blood flow rates in the pulmonary artery (Qp), ascending aorta (Qs), and flow ratio Qp/Qs were determined by PC-MRI with SENSE reduction-factor 2 and 3 (SF-2, SF-3). Additionally, we used PC-MRI with higher spatial in-plane resolution (1.6 x 2.1 mm vs. 2.3 x 3.1 mm) with/without SF-3. Standard (= reference) PC-MRI, which used two excitations (NEX=2), was compared with PC-MRI sequences with NEX=1. RESULTS: Reduction of signal averages and application of SENSE accelerated flow measurements by a factor of 3.8 (5.5) using PC-MRI with SF-2 (SF-3): Scan time was 36 sec (SF-2) and 25 sec (SF-3) at average heart rate of 69/min. The mean Qp/Qs by reference PC-MRI was 1.03 +/- 0.07 (range, 0.89-1.16), and 1.0 +/- 0.11 (range, 0.86-1.24) by PC-MRI + SF-3, respectively. For blood flow rate through the pulmonary artery and aorta, and for Qp/Qs ratio, we found differences of -3% to +4%. The lower limits of agreement (mean - 2 SD) ranged between -14% and -21%, and upper limits (mean + 2 SD) between +9% and +30%, demonstrating clinically acceptable agreement with standard PC-MRI (Bland-Altman analysis). PC-MRI at higher in-plane resolution both with/without SENSE yielded slightly smaller aortic and pulmonary flows (mean differences 5% to 7%, p < 0.05). CONCLUSIONS. In adults, PC-MRI may be safely combined with SENSE to reduce scan time for a quantitative flow measurement in the great arteries to "30 seconds. High in-plane resolution was not advantageous. PMID- 15881516 TI - In vivo detection of encapsulated intracardiac paraganglioma by delayed gadolinium enhancement magnetic resonance imaging. AB - Intracardiac paragangliomas are rare endocrine tumors that are usually benign and can be cured by surgical resection. Both invasive and encapsulated forms have been described and degree of invasion determines surgical complexity. We present in vivo detection of fibrotic encapsulation in a cardiac paraganglioma using delayed enhancement (DE) cardiac magnetic resonance imaging (CMRI), later confirmed on pathology. This finding improved presurgical risk assessment and helped guide management. Tumor necrosis was also easily identified. DE appears useful in the assessment of intracardiac tumor invasion. PMID- 15881517 TI - Adjunctive role of cardiovascular magnetic resonance in the assessment of patients with inferior attenuation on myocardial perfusion SPECT. AB - PURPOSE: Inferior attenuation is a common problem in the interpretation of myocardial perfusion SPECT. We explored whether cardiovascular magnetic resonance (CMR) was a useful adjunct in differentiating between artifactual attenuation of the inferior wall and the presence of myocardial infarction and/or ischemia. METHODS: We used CMR to assess resting wall motion, myocardial perfusion, and the presence of infarction with late gadolinium enhancement in 30 patients with presumed inferior attenuation on ungated myocardial perfusion SPECT, but where uncertainty was present over interpretation of the inferior wall. Perfusion CMR was analyzed visually and quantitatively. RESULTS: In 23 patients (77%), CMR excluded infarction or ischemia in the inferior wall. The myocardial perfusion reserve index (MPR1) was the same in the inferior and remote myocardium (1.74 +/- 0.43 vs. 1.77 +/- 0.50, p = 0.61). Coronary angiography was performed in 11 of these patients, and was normal in all cases. In the remaining seven subjects (23%), significant abnormality was detected by CMR (infarction, 5; wall motion abnormality, 3; perfusion defect, 5). In these patients, the MPR1 was reduced in the inferior myocardium compared with remote (1.07 +/- 0.19 vs. 1.74 +/- 0.49, p = 0.04). Coronary angiography was performed in three of these patients, revealing significant coronary disease in the artery supplying the inferior territory in all patients. CONCLUSION: Approximately one-quarter of patients with inferior attenuation on ungated, nonattenuation corrected myocardial perfusion SPECT have abnormalities on CMR. CMR can readily distinguish between artifact, ischemia, and infarction in these cases and in some cases might obviate the need for diagnostic coronary angiography. PMID- 15881518 TI - Disparity between dobutamine stress and physical exercise magnetic resonance imaging in patients with an intra-atrial correction for transposition of the great arteries. AB - BACKGROUND: In patients with an intra-atrial correction for transposition of the great arteries (TGA) an abnormal response to stress testing is common. However, hemodynamic responses may vary substantially when different stress tests are used. We compared the hemodynamic response to dobutamine stress with the response to physical exercise in patients and controls. METHODS: Thirty-nine patients and 25 age/sex-matched control subjects underwent either dobutamine stress (15 microg/kg/min) or submaximal physical exercise cardiovascular magnetic resonance. End-systolic and end-diastolic right ventricular volumes (ESV; EDV) were determined. Five representative patients underwent both stress tests. For these patients, wall thickening reserve was calculated as systolic wall thickening during stress minus systolic wall thickening at rest. RESULTS: In controls, dobutamine stress and physical exercise showed similar responses: stroke volume, cardiac output, and ejection fraction increased significantly, whereas ESV decreased significantly and EDV was unchanged. In patients, stroke volume did not increase with either dobutamine or exercise (-8.6% vs. 2.9%). Ejection fraction increased significantly with dobutamine (16%, p < 0.001) but tended to decrease during exercise (-2.1%, P = NS). EDV and ESV decreased during dobutamine but were unchanged during exercise. (-22% vs. 5.0%, P < 0.001; -36% vs. 9.0%, P < 0.01 respectively). Wall thickening reserve was higher with dobutamine than with exercise (0.9 mm vs. -0.6 mm, P = 0.02). CONCLUSION: Dobutamine stress and physical exercise cannot be used interchangeably for assessment of systolic and diastolic function in patients with an intra-atrial correction for TGA. This may have consequences for the use of different stress CMR approaches in the clinical setting. PMID- 15881519 TI - Cardiovascular magnetic resonance in endocardial fibroelastosis. AB - Endomyocardial fibroelastosis is a rare disorder marked by characteristic morphological changes of the endocardium. We report a case of endomyocardial fibroelastosis diagnosed by late gadolinium-enhanced CMR and confirmed by endomyocardial biopsy. The patient improved following treatment with imatinib. PMID- 15881520 TI - A new approach for rapid assessment of the cardiac rest period for coronary MRA. AB - BACKGROUND: Effective suppression of cardiac motion is crucial for MR coronary angiography (MRCA). Thus, we evaluated a new technique for rapid and automatic detection of the cardiac rest period in comparison to the conventional visual assessment of the coronary artery rest periods. METHODS: One hundred and thirty five consecutive cardiac patients were examined (Philips Intera CV 1.5 T, Best, The Netherlands). Visual assessment of the left and right coronary rest periods was done using a cine-SSFP scan with a transversal slice orientation (retrospective gating, 40 phases/cardiac cycle); the coronary rest period was defined as the duration of the coronary artery being completely within a region of interest placed on the outer edge of the cross-section of the vessel. Common coronary rest period as determined from visual assessment was defined as the intersection of both coronary artery rest periods. For comparison, an automatic technique was applied: using the position of the shim volume to define a correlation kernel, the cross-correlations of consecutive cine images were registered and displayed in a graph. Based on these cross-correlation values, the cardiac rest period was detected. The correlation between the visual and automated analysis was assessed. RESULTS: A high correlation between the automatically and visually determined starting points for the coronary artery rest periods and the cardiac rest period was found. The automatically assessed cardiac rest period was significantly shorter in comparison to the visually assessed left and right coronary artery rest period (103 +/- 46 ms vs. 158 +/- 72 ms and 117 +/- 52 ms, respectively; p < 0.001). However, the common coronary rest period demonstrated excellent agreement with the cardiac rest period (r = 0.93, p < 0.001) without a significant difference in duration (109 +/- 52 ms vs. 103 +/- 46 ms). CONCLUSIONS: Automated analysis of the cardiac rest period yielded similar results compared to the visual analysis. This rapid assessment of a cardiac acquisition window may be most helpful for MRCA, especially when aiming at 3-dimensional coverage of the whole coronary arterial tree during a single scan. PMID- 15881521 TI - Potential intrinsic error of noninvasive coronary angiography. AB - PURPOSE: With current noninvasive techniques compromises have to be accepted for coronary imaging, e.g., partial coverage of the coronary artery tree. The aim of the study was to estimate the potential intrinsic error of partial coverage from a database of invasive angiograms. METHODS AND RESULTS: The localization and severity of coronary artery stenoses and, if percutaneous coronary intervention (PCI) was performed, the balloon and stent size were extracted from a large database of 21,335 selective coronary angiograms. Of stenoses with >50% diameter reduction, 31.4% were located in distal segments and minor side branches, constituting 28.8% of interventional treatment. In 5% of patients undergoing their first invasive angiogram, the most proximal relevant stenosis was found in a distal segment or minor side branch. Most interventions (83.5%) were performed in main coronary artery segments. CONCLUSION: Coronary artery stenoses were found and interventional treatment performed in all coronary segments. Therefore, noninvasive coronary imaging of only proximal and medial segments and major side branches is an inadequate strategy for complete diagnosis or as a guide to therapeutic decisions. However, the currently available noninvasive techniques allow the detection of relevant stenoses in 95% of patients with suspected coronary artery disease (CAD) to prepare for further invasive diagnostic and therapeutic planning. PMID- 15881522 TI - Reproducibility of black-blood coronary vessel wall MR imaging. AB - PURPOSE: New magnetic resonance imaging (MRI) methods can provide high spatial resolution images of the coronary artery wall, and statistically significant differences in coronary vessel wall thickness between atherosclerotic disease patients and control subjects have been shown. The aim of this study is to assess inter-study reproducibility (i.e., repeatability) of the cross-sectional 2D coronary vessel wall MRI using a black-blood technique. METHODS AND RESULTS: Twelve healthy adult subjects were studied with the use of a commercial 1.5 T CMR system. A double inversion recovery fast spin-echo (FSE) sequence (black blood) was used. Each subject was scanned twice on different days. The mean RCA lumen area difference between study 1 and study 2 was 0.004 mm2, and the coefficient of repeatability (COR) was +/- 0.06 mm2. The mean RCA wall thickness difference between study 1 and study 2 was 0.02 mm, and the COR was +/-0.39 mm. CONCLUSION: Cross-sectional coronary vessel wall imaging with 2D double-IR FSE is highly reproducible for serial evaluation of coronary vessel walls and atherosclerotic coronary artery disease. PMID- 15881523 TI - Drug eluting coronary stent: in vitro evaluation of magnet resonance safety at 3 Tesla. AB - PURPOSE: To evaluate MR safety at 3 Tesla for a drug eluting coronary stent. METHODS: A drug eluting coronary stent (Endeavor, cobalt alloy, Medtronic Vascular, Santa Rosa, CA) was evaluated for magnetic field interactions, heating, and artifacts at 3 Tesla. MRI-related heating was assessed with the stent in a gelled saline-filled phantom using a transmit/received RF body coil with a whole body averaged SAR of 2.0 W/kg. Artifacts were characterized using T1-weighted, spin echo, and gradient echo pulse sequences. RESULTS: The stent exhibited minor magnetic field interactions that will not cause migration. Heating was not substantial (+0.5 degrees C). Artifacts may create a problem if the area of interest is in the same area or close to the stent (e.g., for a T1-weighted, spin echo pulse sequence, within approximately 16 mm; for a gradient echo pulse sequence, within approximately 23 mm). Conclusion. The findings indicated that it would be safe for a patient with this cobalt alloy-based, drug-eluting coronary stent to undergo MRI at 3 Tesla or less. Importantly, because of the relative lack of magnetic field interactions, MRI may be performed immediately after implantation. PMID- 15881524 TI - Stenotic mitral valve prosthesis with left atrial thrombus. AB - We report the CMR findings of a patient with stenotic biological mitral valve prosthesis that was complicated by thrombus formation in the left atrium. The case illustrates the ability of CMR to assess biological prosthetic valves and to provide a comprehensive diagnostic approach in this clinical scenario. PMID- 15881525 TI - New techniques for the reconstruction of complex vascular anatomies from MRI images. AB - The accurate representation of two-dimensional images in three dimensions has become important for many medical imaging applications and for cardiac magnetic resonance imaging (MRI) in particular. Reconstruction methods applied after data acquisition can produce three-dimensional information from two-dimensional data and make applications such as surgical planning more effective. Current reconstruction techniques usually demand contrast agents, and can suffer due to poor segmentation and sampling constraints that cause surface irregularities and distort dimensions. The novel technique presented here for anatomical modeling uses adaptive control grid interpolation (ACGI) to approximate data not captured by scanning, and a progressive shape-element segmentation technique to complete reconstruction. Quantitative validations conducted on models of pediatric cardiac malformations have confirmed the theoretical advantages of this technique, and that higher quality is achieved than with competing methods based on geometric parameters. Vascular diameters from reconstructions showed errors of less than 1% for a known geometry as compared to over 9% for competing methods. Qualitatively, models produced with the new methodology displayed substantial improvement over alternatives. Approximately 50 rare cardiac structures, including surgically altered Fontan and atypical aortic anatomies, have been reconstructed. All data used to create these reconstructions were acquired using standard pulse sequences and without contrast agents. Benefits of the new technique are particularly evident when complex vascular configurations complicate reconstruction. The proposed methodology enables a powerful tool allowing physicians to analyze and manipulate highly accurate and clearly presented vascular structures in an interactive medium. PMID- 15881526 TI - Application of spatial modulation of magnetization (SPAMM) to children: the effect of image resolution on tagging pattern. AB - BACKGROUND AND OBJECTIVE: Spatial modulation of magnetization (SPAMM) is a valuable magnetic resonance imaging technique for studying ventricular biomechanics. In order to track the intersection points of the stripes to calculate regional wall motion and strain, the stripe spacing should be at most half the wall thickness, yet sufficiently larger than the image pixel size in order that the stripes be well resolved. These conflicting requirements, that the grid spacing be much smaller than the wall thickness yet much larger than the pixel size, are relatively easy to meet in adult subjects but are difficult in children because of their small size. The purpose of this study was to delineate the effect of pixel size relative to SPAMM grid spacing on the stripe pattern produced by SPAMM with application towards its use in children and to present a new approach to the analysis of these images. METHODS: We performed SPAMM imaging on a 1.5 Telsa Siemen's Vision MR system on a phantom, using an artificial ECG (R R interval = 450 ms for triggering), holding the pixel size constant and varying the degree of stripe spacing. We used both square (1 mm) and rectangular (1 mm by 2 mm) pixels. We express the ratio of grid to pixel size as the ratio of the center-center spacing of the grid lines to the horizontal pixel size. We retrospectively reviewed the SPAMM images on 10 patients with a ratio of grid to pixel size approximately 4:1 and 10 with a ratio of grid to pixel size approximately 8:1. We further performed SPAMM imaging in four patients with different grid to pixel size ratios in the same patient. Finally, we tested a new algorithm to track the signal intense regions rather than the signal poor intersection points of the grid lines, which were compared on three ventricles with SPAMM tagging. RESULTS: In a phantom, the effect of decreasing the separation between stripes while keeping the resolution of the image constant changed the stripe pattern from a series of two parallel lines perpendicular to each other to a "checkerboard" pattern. With a relative grid:pixel ratio of 8:1 as used with adult studies, the dark bands and the crossing points are well defined. As the ratio decreases from 8:1 to 6:1, the black band is less well resolved. When the resolution is reduced further to a grid:pixel ratio of 4:1, the image appears to be a checkerboard of white and dark squares. This occurred with both square and rectangular pixels. The effect in vivo is similar. When the ratio is approximately 8:1, all patients demonstrated a stripe pattern as a set of parallel lines perpendicular to each other. When the ratio was approximately 4:1, all patients demonstrated the stripe pattern as checkerboard. This was found to be the case in the same patient, whether varying the pixel size by changing the field of view or matrix or by changing the grid spacing. We also found that tracking the signal intense regions was equivalent to tracking the signal poor intersection points of the grid, and this approach was much easier to implement. CONCLUSION: With decreasing ratio of grid spacing to pixel size, SPAMM stripe patterns change from a set of parallel lines perpendicular to each other to a "checkerboard" pattern. This effect has implications for tracking techniques to determine strain and wall motion. At smaller ratios, as is needed sometimes in children, it is easier to track the signal intense regions rather than the "intersection" points of the stripes. Both these approaches to tracking are equivalent. PMID- 15881527 TI - Impact of the ECG gating method on ventricular volumes and ejection fractions assessed by cardiovascular magnetic resonance imaging. AB - PURPOSE: Most MRI centers currently use prospective ECG triggering and fast gradient-echo sequences for image acquisition. Retrospectively gated sequences allow the coverage of the entire cardiac cycle. There is concern about whether ventricular volumes and ejection fraction (EF) differ according to the gating method used for image acquisition. We sought to evaluate the impact of the gating method on measurements of right and left ventricular volumes and EF in normal subjects. MATERIALS AND METHODS: Fifteen subjects with no cardiovascular disease were investigated by MRI using a 1.5 Tesla scanner. Images were acquired with a gradient-echo sequence with steady-state free precession (SSFP) using the standard short-axis method for volume and EF measurements. Images were acquired with 6-mn-thick slices using both prospective triggering and retrospective gating. Left and right ventricular volumes (EDV, ESV, SV) and EF were determined with a commercially available software package (Argus, Siemens). RESULTS: EDV and SV calculated from short-axis images were significantly smaller with the prospectively triggered SSFP sequence (mean difference: EDV left: 13.9 +/- 4.4 mL, p < 0.0001; SV left: 13.5 +/- 4.8 mL, p < 0.0001; EDV right: 14.2 +/- 3.9 mL, p < 0.0001; SV right: 14.7 +/- 5.9 mL, p < 0.0001). EF was significantly smaller for the right ventricle (mean difference -3.6 +/- 3.3%, p = 0.0008) and the left ventricle (mean difference -2.3 +/- 3.3%, p = 0.02). ESV remained unchanged (mean difference: ESV left: 0.47 +/- 3.5 mL, p = 0.6179; right ESV: 0.5 +/- 3.7 mL, p = 0.6083). CONCLUSION: The gating method has a significant impact on volume and EF measurements. The global ventricular EF is underestimated by using the prospective triggering technique. However, the difference in the left ventricle is small and might not be of clinical relevance. PMID- 15881528 TI - Operator induced variability in cardiovascular MR: left ventricular measurements and their reproducibility. AB - PURPOSE: To assess the intra- and inter-operator variability of the manual planning of cardiovascular magnetic resonance imaging and to evaluate the influence of these factors on the functional parameters of the left ventricle (LV). METHOD: The study population consisted of 10 healthy volunteers. For each subject the manual planning of the short-axis cine acquisitions was carried out twice by one operator and once by a second operator. Left ventricular volume, mass, and function were manually evaluated twice by one experienced observer, resulting in an approximation of the intra-observer variability factor. The intra and inter-operator variation factors were estimated as the difference between the total and intra-observer variation components. RESULTS: LV end-diastolic volume varied by 3.3% and 4.16%, and LV end-systolic volume by 5.84% and 6.23% for intra- and inter-operator studies, respectively. The variability for LV mass at end-diastole was equal to 4.23% in both studies. For the ejection fraction the variability was 3.56% and 2.97% for intra- and inter-operator studies, respectively. Comparison of reproducibility between intra- and inter-operator studies resulted in insignificant statistical differences. Bland-Altman limits of agreements revealed no systematic bias in differences between measurements with respect to their means. Reliability of the planning expressed as the angular deviation of the short-axis imaging planes amounts to 2.67 -/+ 1.5 degrees and 4.99 +/- 2.17 degrees for the intra-operator and inter-operator studies, respectively. For EDV, ESV, and EF approximately 75-80% of the total variation can be explained by the within or between operator variation, while the same percentage is 60% for LVM. CONCLUSIONS: Our study confirms the excellent inter- and intra-operator reproducibility of the cardiovascular magnetic resonance measurements of the left ventricular volumes and mass in a group of healthy volunteers. PMID- 15881529 TI - The angiotensin II type 2 receptor and improved adjacent region function post-MI. AB - Angiotensin II type 2 receptor (AT2-R) overexpression in the mouse heart preserves left ventricular (LV) size and global LV function during post-MI remodeling. We hypothesized that CMR tagging would localize regional improvements in myocardial function during post-MI remodeling in AT2-R cardiac overexpressed transgenic mice (TG), which could explain the preservation of global LV function post-MI. Six male wild-type (WT) C57BL/6 mice and 10 TG mice were studied by CMR at baseline (day 0) and days 1, 7, and 28 post-MI. MI was induced by 1 hour occlusion of the LAD followed by reperfusion. On day 1 post-MI, gadolinium-DTPA was injected to assess infarct size. LV size and function was assessed by cine CMR. Mean % circumferential shortening (%CS) was calculated within infarcted, adjacent, and remote regions at each time point in WT and TG mice. Quantitative interstitial collagen and mean myocyte cross-sectional area was measured postmortem at day 28 post-MI. LV end-systolic volume was lower and ejection fraction higher at baseline in the TG group and these differences were maintained post-MI. Within infarcted and remote zones, although %CS was higher in TG mice at day 0, there was no difference by day 28 between groups. Within adjacent regions, while there was no difference at day 0 or 1 in TG vs. WT, %CS was significantly higher in TG mice by day 7, and these changes persisted out to day 28 post-MI. Regional interstitial collagen and myocyte size were similar between groups. Thus, myocardial tagging can detect regional differences in contractile function post-MI in TG mice, and AT2-R overexpression is associated with improved contractile function in adjacent noninfarcted myocardium. PMID- 15881530 TI - Atrial and ventricular volume and function in persistent and permanent atrial fibrillation, a magnetic resonance imaging study. AB - Left atrial size is independently related to cardiovascular morbidity and mortality, and atrial fibrillation (AF) is strongly associated with atrial size. Our aims were to report atrial and ventricular dimensions in patients with AF evaluated with magnetic resonance imaging (MRI), and to assess the inter-study reproducibility of the measurements. Nineteen healthy volunteers, 19 patients with permanent AF, and 58 patients with persistent AF had cardiac dimensions evaluated by 6-mm cinematographic breath-hold MRI scans using a 1.5 Tesla Siemens Vision Magnetom scanner with a phased array chest coil. Intraobserver variability and inter-study reproducibility of the cardiac volumes and ejection fractions (EF) gave acceptable Bland-Altman plots, good correlations (R2: 0.80-0.99), and low reproducibility coefficients. The mean atrial volumes were similar in the two groups with AF [systolic vol. index (SVI): 75.9-80.3 mL/m2; diastolic vol. index (DVI): 77.4-82.1 mL/m2] and significantly different from the healthy volunteers (SVI: 30.3 mL/m2; DVI: 62.3 mL/m2; p < 0.0001). Mean left ventricular (LV) volumes and EF were significantly different in permanent AF (SVI: 34.2 mL/m2; DVI: 68.3 mL/m2; EF: 50.8%) compared to persistent AF [SVI: 44.0 mL/m2 (p = 0.02); DVI: 77.2 mL/m2 (p = 0.03); EF: 44.9% (p = 0.02)], and closer to the normal values (SVI: 22.4 mL/m2; DVI: 66.5 mL/m2; EF: 67.0%). MRI is a highly reproducible method for measurement of atrial and ventricular dimensions in healthy volunteers and in patients with AF. Our results suggest that atrial dilatation appears within the first months of AF and stays more or less unchanged thereafter. The LV appears to dilate early as a response to AF, but later seems to adapt. PMID- 15881531 TI - The apparent inversion time for optimal delayed enhancement magnetic resonance imaging differs between the right and left ventricles. AB - BACKGROUND: Delayed post-contrast magnetic resonance (MR) imaging involves suppression of signal from myocardium using inversion times (TI) between 150-225 ms, when the myocardium appears dark and fibrotic scar appears bright. We noticed that at a TI optimized for signal suppression of the left ventricle (LV), the right ventricle (RV) appeared brighter. PURPOSE: The purpose of this study was to evaluate the TI for signal suppression in RV compared to LV, and to try and identify the cause of this observation. Methods. We studied 31 patients (ages ranged from 17-79 years, 11 females) who had an MR scan on a 1.5 T GE scanner. Delayed post-contrast short-axis images were obtained 20 minutes after injection of 0.2 mmol/kg of intravenous gadolinium chelate. TI optimization was performed by acquiring a range of TI times within a single breath hold, in increments of 25 msec. The TI time that resulted in lowest signal for the RV arid LV was recorded. RESULTS: With the imaging sequence employed, the TI leading to LV signal suppression ranged from 150-225 ms. At the TI that resulted in LV signal suppression, the corrected signal from the RV was significantly higher as compared to the LV (29 +/- 13 au vs. 15 +/- 8 au, p < 0.001). The findings were similar using only the body coil. The TI required to suppress the RV was usually < or =150 msec. The observation persisted before and after gadolinium infusion. CONCLUSION: The TI for myocardial signal suppression appears to be different between LV and RV. Potential mechanisms include partial volume averaging with fat or blood pool (related to increased trabeculation) in the RV. Alternatively, increased blood pool signal (within Thebesian veins or arterioluminal communications) in RV compared to LV leads to altered TI times due to similar partial volume effects. PMID- 15881532 TI - Standardizing the definition of hyperenhancement in the quantitative assessment of infarct size and myocardial viability using delayed contrast-enhanced CMR. AB - PURPOSE: To evaluate a standardized definition of delayed hyperenhancement in the analysis of contrast-enhanced cardiac magnetic resonance (ceCMR) imaging. PATIENTS AND METHODS: CeCMR was performed in 15 patients with chronic ischemic heart disease. Delayed hyperenhancement was analyzed both by visual analysis by an experienced team of observers, and after thresholding the window setting of the images at 2, 3, 4, 5, and 6 SD above the mean signal intensity of remote, normal myocardium in the same slice. In each patient, total infarct size (TIS) and segmental infarct extent (SIE) were calculated. RESULTS: TIS and SIE were 22.9 +/- 12.2 mL and 32 +/- 28% after visual analysis. Thresholding the window setting at 2, 3, 4, and 6 SD above signal intensity of remote caused a 40%, 31%, and 17% increase (p < 0.007) and a 7% decrease (p = NS) in TIS, and a 75%, 41%, and 16% increase and 22% decrease in SIE (p < 0.001), respectively. There was no difference between visual analysis and analysis after thresholding at 5 SD. CONCLUSION: Analyzing ceCMR with a standardized definition of hyperenhancement related to the signal of remote, nonenhanced myocardium may result in considerable overestimation of infarct size at the usual cut-off of 2 SD. PMID- 15881533 TI - A fast and effective method to assess myocardial necrosis by means of contrast magnetic resonance imaging. AB - PURPOSE: Contrast magnetic resonance (CMR) can identify myocardial necrosis after gadolinium administration as a hyperenhanced (HE) area. Yet there are no software tools that can effectively quantify such an area. The aim of this study is to develop a robust and effective algorithmic method for defining the extent of myocardial necrosis evidenced through CMR. METHOD: Fifteen patients with previous myocardial infarction underwent nitrate Tetrofosmin G-SPECT and CMR. A software tool was developed, allowing semiautomatic detection of endocardial and epicardial borders and the automatic detection of HE regions. The accuracy of the proposed quantitative method of analysis has been tested with G-SPECT analysis that it is less than an ideal method for assessing myocardial viability, but at present is accepted and widely used in the clinical arena. RESULTS: Segmental (SEHE) and global extension of HE were evaluated. HE was present in 161 of the 255 analyzed segments. Of the 161 HE segments, the mean SEHE was 36 +/- 30%. The operator independence (intraobserver: r = 0.97, p < 0.0001, interobserver: r = 0.95, p < 0.0001) was good and significant, with noticeable time savings with respect to manual analysis. There was strong and inverse correlation between SEHE and scintigraphic regional uptake reduction (r = -0.66, p < 0.0001), and also a positive correlation between SEHE and SPECT defect extension (r = 0.75, p < 0.0001). When assessing the global extent of necrosis, the correlation between the two techniques was strong (r = 0.79, p = 0.0004). CONCLUSIONS: The proposed method of quantifying myocardial necrosis by CMR is highly reliable, reproducible, and operator-independent for quantifying. PMID- 15881534 TI - Heart involvement in T cell lymphoma through hypereosinophilic syndrome: a common complication of a rare condition. AB - This case describes a 42-year-old male affected by hypereosinophilic syndrome associated with angioimmunoblastic lymphoma. Heart involvement was suspected at ECG mimicking left ventricular hypertrophy. MRI clarified the extensive endomyocardial fibrosis, confirming the role of this technique in in-vivo tissue characterization. Finally, the study investigates the association of T cell lymphoma, hypereosinophilic syndrome, and Loeffler endomyocardial disease. PMID- 15881535 TI - Assessment of myocardial function: a review of quantification methods and results using tagged MRI. AB - Tagged MRI provides a noninvasive way to assess the regional function of the heart. Clinical use of myocardial strain measurements from tagged MRI requires identifying new normative values. As for cardiac motion estimation, a variety of methods for quantifying myocardial deformations have been proposed in the image analysis and medical literature, based on heart geometry and continuum mechanics. This article comparatively reviews existing quantification techniques, and synthesizes their results to establish confidence intervals for the standard deformation parameters. PMID- 15881536 TI - Simultaneous occurrence of two different primary cardiac tumors in an 84-year-old woman characterized by cardiovascular magnetic resonance imaging. PMID- 15881537 TI - Delayed contrast-enhanced magnetic resonance imaging for the detection of autoimmune myocarditis and long-term follow-up. AB - MRI proved to be a valuable tool for the evaluation and monitoring of myocarditis. We report the case of a 36 year old caucasian male with an undifferentiated collagenosis who presented first four years ago with unspecific symptoms and impaired exercise capacity. On echocardiography left ventricular function was impaired as well as on MRI. In addition, after administration of Gd DTPA an intramyocardial pathological signal enhancement was observed on TSE T1 weighted and contrast enhanced FLASH 3D IR sequences. Based on several diagnostic tests including myocardial biopsy an autoimmune myocarditis due to an undifferentiated collagenosis was diagnosed. On long time follow up over almost 3 years with repeated MRI examinations, the delayed hyper enhancement (dHE) decreased or disappeared with successful treatment, respectively. However, in the area of recurrent inflammation, a persisting area of dHE developed with a subsequent perfusion defect. This area represents myocardial fibrosis due to recurrent inflammation. PMID- 15881538 TI - The assessment of left ventricular (LV) function plays a central role in cardiac imaging. PMID- 15881539 TI - Work-related homicides among seafarers and fishermen. AB - OBJECTIVE: To analyse work related homicides among various defined populations of seafarers and fishermen during the period 1976-2002, to compare homicide rates with other occupations and general populations, and to discuss implications for maritime health and safety. METHODS: A longitudinal study of work related homicides, based on official mortality files, with an aggregated population of almost two million seafarer-and fishermen-years at risk. RESULTS: Of 46 work related homicides, 44 occurred among seafarers, and two among fishermen. Homicide rates were 1.5 per 100,000 worker-years among seafarers in British merchant shipping (1976-2002), 2.9 per 100,000 among British seafarers employed in non British shipping (1986-95), 3.7 in Singapore shipping (1984-95) and in Hong Kong shipping (1981-95), and 0.4 among fishermen in British fishing (1976-2002). The homicide rate in British shipping increased from 2.7 per 100,000 seafarer-years in 1961-65 to 4.4 per 100,000 in 1971-75, but has declined to 0.0 in 1996-2002. CONCLUSIONS: Although work related homicides were generally quite rare occurrences, among British seafarers and fishermen, they were largely caused by attacks from assailants, other than colleagues, that occurred ashore. The decline in the homicide rate in British shipping since the 1970s has coincided with the reduction in deep-sea ships trading to high risk countries for homicide. Among Asian seafarers, most homicides resulted from disputes among colleagues that mainly occurred on board ships. Prevention should include alertness to potential risks when spending off-duty time ashore in locations with high homicide rates, and conflict resolution for disputes among seafarers. PMID- 15881540 TI - Dealing with gastrointestinal illness on a cruise ship--Part 1: Description of sanitation measures. Part 2: An isolation study. AB - BACKGROUND: Recent outbreaks of gastrointestinal illness (GI) on passenger ships have caused cruise companies to expand their sanitation programs. OBJECTIVES: To describe sanitation procedures and measures involving the medical staff of one cruise ship (Part 1), and to show how one of the new measures, isolation, influenced medical practice (Part 2). METHODS: Consecutive GI logs were reviewed from 17 cruises of a ship with an average of 590 passengers and 611 crew per day. RESULTS: During a 250-day period 207 persons were isolated for a total of 419 days: 113 passengers (75 with GI, 38 asymptomatic contacts) and 94 crew (57 with GI, 37 contacts). The percentage of GI cases per 7 days varied between 0.10% and 0.43% among passengers and between 0 and 0.39% among crew, except for one cruise when the percentage per 7 days reached 2.16% among passengers and 0.61% among crew. CONCLUSIONS: A detailed operational directive for all aspects of sanitation is helpful for prevention and control of GI outbreaks on ships. A strict isolation policy is an expensive, but critical measure, which in particular challenges the medical staff. PMID- 15881541 TI - Overview of the diseases incidence among seafarers in Japan in 1986-2000: general trends. PMID- 15881542 TI - A survey on the influence of personality on successful career development of the Maritime Academy students. AB - The present research focused on the exploration of those psychological functions that might be helpful in predicting successful career development as prospective seafarers among students of the Maritime Academy (MA) in Gdynia. The study group comprised of 334 MA students. The survey explored intellectual and cognitive ability, selected personality and temperamental traits, social behaviour style, ability to cope with difficult situations and immunity to stress. PMID- 15881543 TI - Diseases of seafarers in Vietnam preliminary report. AB - The authors examined groups of Vietnamese seafarers and shore workers in the port of Haiphong, recorded diseases prevailing among them, and compared the morbidity in these two groups of workers. They also measured the noise level on ships. This is a preliminary report of the work conducted by the staff of the National Institute of Maritime Medicine aimed at better protection and promotion of health of national seafarers. PMID- 15881544 TI - Occupational allergies in the seafood industry--a comparative study of Australian and South African workplaces. AB - Although seafood allergy due to ingestion is commonly observed in clinical practice, the incidence of seafood allergies in general and more specifically in the occupational setting in Australia is largely unknown. The work practices, occupational health services and allergic health problems in 140 seafood processing workplaces in Australia were examined and compared to previous studies in South Africa. A cross-sectional employer-based survey design was used to conduct the study in both countries. In the South African study a response rate of 60% (n = 41) was obtained, compared to a response rate of 18% (n = 140) in Australia. The most common seafood processed by workplaces in South Africa was finfish (76%) and rock lobster (34%). Similarly in Australia, finfish (34%) was the most frequently handled seafood. However, processing of prawns (24%) and oysters (21%) was more common in Australia. Common work processes in South Africa involved freezing (71%), cutting/filleting (63%) and degutting (58%) procedures. Similar processes were followed in Australian industries with the exception of shucking of oysters, particularly common in the aquaculture industries. About half of the workplaces in both countries provided an occupational health service and medical surveillance of workers. However, none of the workplaces in South Africa and only 9% of the workplaces in Australia had industrial hygiene programs for seafood aerosols in place. In both countries positive trends were observed between the size of the workforce and the provision of occupational health services (p<0.005). Similarly, skin rash accounted for highest of all reported health problems (78-81%) followed by asthmatic symptoms (7-10%) and other non specific allergic symptoms (9-15%) in both countries. Most workplaces reported the annual prevalence of work-related symptoms to be less than 5%. In Australia 7% of respondents in workplaces reported workers having left their workplace due to work-related allergic problems. Despite a low response rate of contacted companies in Australia, there were great similarities between the two countries suggesting that there is a significantly elevated prevalence of work related allergic symptoms in both countries. Unexpectedly, mollusc processing was more common in Australia although the occupational health related effects among exposed workers has previously not been investigated in detail and merits further study. It is recommended that further epidemiological studies focus on seafood exposure in Australia and identify specific risk factors for sensitisation. PMID- 15881545 TI - Assessment of clinical course and outcome of Plasmodium falciparum malaria in Angola diagnosed by microscopic and molecular methods. AB - Malaria still remains the main infectious cause of morbidity and mortality in the world. The majority of cases occur in tropical Africa, including Angola. The study was performed in Angola, in the Health Centre of St. Lucas in Kifangondo, near Luanda during the dry season, from May to October 1999. 168 patients with symptoms of malaria were referred for study enrolment. In the study we assessed the efficacy of treatment in falciparum malaria based on clinical, microscopic and polymerase chain reaction (PCR) examination, and estimated the prevalence of mutations in the pfcrt gene in the P. falciparum isolates from chloroquine treated patients. In 159 (95%) patients P. falciparum malaria was diagnosed using microscopic methods. This was confirmed by PCR. We admitted 68 patients to the hospital; 13 patients died from complicated malaria. We have analysed 59 cases of P. falciparum malaria known to be chloroquine-treated. In 58 (98%) cases we have found P. falciparum isolates that carried gene polymorphisms typical for chloroquine resistance. In the group with such P. falciparum isolates, 8 people were treated successfully. Reasons for this response are discussed. PMID- 15881546 TI - The implementation of factor analysis for the evaluation of selected blood parameter changes induced by hyperbaric exposure. AB - This paper discusses the application of factor analysis when used to compare selected blood parameter (a three-parameter smear, hematocrit, C3c, C4, IgG, IgA, IgM, CRP, fibrinogen and the level of factor XII) properties, just before, and after exposure to pressure changes, and 24-hours after the completion of decompression. To-date the most popular method of statistical analysis was based only on investigation of the significance of the separated individual parameters. This factor analysis that has not been applied previously in the analysis of such problems, enabled the neutral hierarchic evaluation of the significant parameter changes within their chosen range, and mutual relationships. It seems that the application of this method is purposeful and it can be an objective tool for evaluating the significance of changes in blood constituency induced by pressure. PMID- 15881547 TI - Delayed treatment of bubble related illness in diving--review of standard protocol. AB - The basic treatment of diver with bubble related illness consists of recompression in medical hyperbaric facility. However transportation of injured diver to hyperbaric chamber can last for several hours. During that time the process induced by gas bubbles spread out and finally result in activation of many pathophysiological events. Currently approved standard of treatment before starting of recompression consists of normobaric oxygenation, intravenous or oral fluids and general stabilization of the patient condition. Usage of several pharmacological agents is promising, including corticosteroids, antiplatelet and anticoagulant therapy or lidocaine. Those drugs are most often used in some centres but still there is lack of randomized controlled studies concerning their efficacy in decompression illness of divers. The review of available bibliography presented in this paper leads to conclusion that recommendations of the Second European Consensus Conference on Hyperbaric Medicine "The Treatment of Decompression Accidents in Recreational Diving" published in 1996 in Marseille, France for fluid replacement and drug therapy for decompression accidents are still valid. This protocol includes the fluid treatment, normobaric oxygen and intensive therapy. Other drugs (aspirin, lidocaine, heparin, steroids, calcium channel blockers, antioxidants) should still be treated as an option considered by clinician, but without strong evidences from clinical studies. PMID- 15881548 TI - A European code of good practice for hyperbaric oxygen therapy. AB - In 2001 the Working Group (WG) "Safety" was created within the European COST Action B14 "Hyperbaric Oxygen Therapy" with the main objectives to elaborate recommendations of good practice for hyperbaric medicine and to follow the European normalisation process of hyperbaric chambers. During three years of preparation of the European Code of Good Practice (ECGP) for HBO, the relevant documents concerning safety in hyperbaric chambers from each European country have been revised. The initial document drew on the BHA "Health and Safety for Therapeutic Hyperbaric Facilities: A Code-of Practice" (2000), and later on it was modified using national regulations and standards (from Belgium, Finland, France, Germany, Greece, Italy, Portugal, and Spain), as well as European Norms and existing experience from experts of hyperbaric centres, committees, professional and scientific associations. The ECGP for HBO consists of chapters dedicated to staffing (including responsibilities, competencies and education, minimum team during hyperbaric sessions, fitness and health surveillance), equipment, gas supply, risk management and procedures (including standard and emergency operating procedures, maintenance, record keeping, and patient safety). It also includes ECHM Educational and Training Standards for the Staff of Hyperbaric Centres (1997), ECHM Recommendation for Safety in Multiplace Medical Hyperbaric Chambers (1998), as well as COST B14 Working Group "Technical Aspects" Final Report (2001) including a risk analysis conducted specifically for therapeutic hyperbaric facilities. Many efforts have been spent to make the ECGP for HBO compatible with the new project of the European Norm prEN 14931 "Pressure vessels for human occupancy (PVHO)--Multiplace pressure chamber system for hyperbaric therapy--Performance, safety requirements and testing", which has been prepared at the same time by the CEN/BT/TF 127. Both groups (CEN/BT/TF 127 and COST B14 WG "Safety") cooperated extensively in the field of safety aspects of using hyperbaric chambers as medical devices, and both final versions of documents include many cross references. Being prepared by international Working Group "Safety" and after acceptation by all members of the COST B14 action, the ECGP for HBO represents a harmonised European view on safety in therapeutic hyperbaric facilities and can be used as a reference document for European countries for Guidelines, Regulations, and Standards in hyperbaric medicine. PMID- 15881549 TI - Maritime medicine in France. PMID- 15881550 TI - Psychosocial hazards and seafarer health: priorities for research and practice. AB - This paper discusses the changing nature of work and the emergence and prevalence of psychosocial hazards. It addresses their effect on worker health with particular emphasis on the issue of work-related stress. It then considers how these hazards affect seafarer health in the light of current working conditions in the maritime sector. Finally, specific recommendations are made for future directions for research and practice in relation to these issues. PMID- 15881551 TI - Leishmaniasis among soldiers of stabilization forces in Iraq. Review article. AB - The results of studies concerning the incidence of leishmaniasis among soldiers of Stabilization Forces serving in Iraq in the years from 2003 to March 2004 are presented in this article, in the light of a possibility of importation of this disease to the countries of their origin. Epidemiology, clinical picture, laboratory diagnosis, treatment and prevention of leishmaniasis are described. In mid-2004, more than 2400 Polish soldiers served in Iraq. By the end of June 2004 no case of leishmaniasis was diagnosed among them. Among American troops serving in Iraq, 653 cases of cutaneous and 2 cases of visceral leishmaniasis were reported by the end of March 2004. According to the U.S. sources, the number of infected American soldiers could have been higher, from 750 to 1250 or even more, what made up to nearly 1% of U.S. troops serving in Iraq in 2003-2004. PMID- 15881552 TI - Social status versus formal rank of medical and other officers--an informal survey among passengers and seafarers on cruise ships. AB - BACKGROUND: Modern cruise ships have a rigid hierarchic structure, but status of the officers may have changed over time. OBJECTIVES: To get an impression of how cruise officers and crew (=staff) and experienced passengers perceive the status of various shipboard positions, particularly Doctor and Nurse, and compare with present ranks. METHODS: Passengers and staff during a World Cruise (Ship 1) and staff on a sister-ship (Ship 2) during a voyage without passengers were asked by questionnaire to rank shipboard jobs by assigning 0-4.5 stripes to each position according to perceived social status. Median values were reported. RESULTS: 29% of 109 passengers and 11% of 899 staff responded. Captain was assigned 4.5 stripes by both female and male passengers and staff, followed by Vice Captain (4 stripes, as now, by all). Chief Engineer (4 stripes) was downgraded by male passengers and female staff. Hotel Director (4 stripes) was rated as now by all but male passengers. Executive Housekeeper (2.5 stripes) was upgraded by all but male staff. Doctor (3 stripes) received 4 stripes from passengers and 3.5 from staff. Nurse (2 stripes) got 3 stripes from passengers and 2 stripes from staff. All upgraded Security Officer (2 stripes). CONCLUSIONS: The participants seemed to accept the hierarchic rank structure aboard, with the captain alone on top. Marine officers, especially those considered responsible for safety, security and health, were ranked high by all, while passengers also tended to upgrade hotel officers with visible problem-solving positions. Passengers and staff agreed that the Doctor and the Security Officer deserve more stripes, while the Nurses were upgraded by passengers, but not by staff. PMID- 15881553 TI - St. Lucas Heath Center in Kifangondo, Angola. PMID- 15881554 TI - Actual problems of the Russian Maritime Health Services. PMID- 15881555 TI - Alzheimer's disease--time to act is now. PMID- 15881556 TI - Descriptive study of chronic calcific pancreatitis in Sri Lanka. AB - OBJECTIVES: To describe the potential risk factors, clinical features, biochemical and radiological features, and management of chronic calcific pancreatitis. DESIGN: Cross-sectional descriptive study. SETTING: Tertiary care general hospital. PATIENTS: Fifty patients with pancreatic calcification referred to the Colombo South Teaching Hospital, and 50 age-matched controls from healthy relatives or friends of the patients. MEASUREMENT: Height and weight measurements, immunoreactive insulin levels and trypsin levels of duodenal aspirates were estimated. Plain abdominal xray and ultrasonography were performed. INTERVENTION: Endoscopic retrograde cholangiopancreaticography (ERCP) was attempted on all patients during which duodenal aspirates were collected. Success rates of ERCP and response to endotherapeutic procedures were recorded. RESULTS: Twenty two of the 50 chronic calcific pancreatitis (CP) patients were diagnosed to have chronic alcoholic calcific pancreatitis (CACP). Mean age of the CACP patients was significantly higher than that of the chronic calcific pancreatitis of the tropics (CCPT) patients. Severe malnutrition (BMI < 20), frequent consumption of Manihot esculenta (manioc, cassava) and a high consumption of chilli or pepper were identified as possible risk factors for both alcoholic and non-alcoholic CP. Onset of diabetes occurred at a much younger age in the CCPT group than in the CACP group. Mean serum insulin was significantly higher in the CCPT group than in the CACP group and duodenal trypsin level was significantly lower in the CCPT than in CACP group. CONCLUSIONS: Our results confirm the existence of both alcoholic (CACP) and non-alcoholic (CCPT) types of chronic calcific pancreatitis in Sri Lanka. A larger study is required to confirm the associated risk factors such as Manihot esculenta and foods with a high content of chilli or pepper. PMID- 15881557 TI - Adaptation and validation of the parent-child conflict tactics scale for use in Sri Lanka. AB - OBJECTIVE: The purpose of the study was to validate the Parent-Child Conflict Tactics Scale (CTSPC) in Sri Lanka, for use with 12-year old children. DESIGN: A Delphi Process determined the consensual and content validity of the CTSPC. Test retest reliability and internal consistency were determined by a large and a small group administration. SETTING: Professionals from the social and behavioural sciences were the judges in the Delphi Process. Conveniently located schools in the Gampaha District were chosen for determining the test-retest reliability and internal consistency. MEASUREMENTS: This included original CTSPC and its Sinhala translation, a structured interview schedule and a focus group guide. RESULTS: The Sinhala version of the CTSPC showed adequate consensual and content validity. Its test-retest reliability and internal consistency were satisfactory. The instrument is best administered in small rather than in large groups of children. CONCLUSION: The Sinhala version of the CTSPC is appropriate to be used with 12-year old Sinhala speaking schoolchildren in Sri Lanka. PMID- 15881558 TI - Three decades of the Ceylon Medical Journal--analysis using MEDLINE (PubMed). AB - INTRODUCTION: The Ceylon Medical Journal (CMJ) is the only Sri Lankan medical journal that is indexed in MEDLINE (PubMed). Publications in the CMJ from 1965 March to 2001 December were analysed using the PubMed interface of MEDLINE. METHOD: Using PubMed we downloaded the CMJ bibliography in MEDLINE format. Important tagged fields were written to a Microsoft-Access database using a PubMed Grabber/Analyser program. The analysis was done using Access-SQL and PubMed queries. RESULTS: There were 1472 citations by 1373 authors. 944 authors had only one and 176 had two articles. The top 10 authors contributed 8% and the top 25, 15.4% of the articles. Publications types were: 68.5% 'journal articles', 10.4% letters, 4.2% historical articles and 4.1% reviews. Controlled clinical trials, randomised controlled trials and clinical trials together totalled only 39 (2.1%). Articles were classified using 2 to 44 medical subject headings (MeSH; average 11.4, mode 9) from the MeSH vocabulary. CMJ articles classified using broad MeSH categories were (top five): infections 370(15 %), pathological conditions signs and symptoms 266( 10.8%), haemic, lymphatic and immunologic 199 (7.7%), endocrine, nutritional and metabolic 189 (7.7%), neoplasms 179(7.3%). DISCUSSION: For 938 (68%) authors, publishing in the CMJ was a one time affair. The top 50 authors contributed nearly a quarter (23.2%). A product of this research is an off-line CMJ searching system from 1965 to 2001 with menu driven search facilities which will be a useful tool for researchers. PMID- 15881559 TI - A preliminary study of the hospital-admitted older patients in a Sri Lankan tertiary care hospital. AB - OBJECTIVE: To provide reasons for admission, comorbidities, physical and psychosocial characteristics, and outcome of older patients admitted to hospital in Sri Lanka, and to assess adequacy of routine assessment. DESIGN: Descriptive study. SETTING: University Medical Unit, National Hospital of Sri Lanka during 10 weeks commencing August 2003. PATIENTS: All > or = 65 years admitted to the unit, excluding transfers for specialised management and direct admissions by consultants or clinics; 75 men and 75 women. MEASUREMENTS: Vision, hearing and speech, tests for tendency to fall, basic activities of daily living (BADL), mood and cognition, body mass index and waist circumference. RESULTS: Seventy three per cent and 40% had cognitive dysfunction and depression., 20% required assistance in > or = 5/9 BADL, 23% had unprovoked falls within past year, and 57% had a tendency to fall, 34% had < or =6/60 vision. CONCLUSIONS: Falls, depression, cognitive dysfunction and severe visual impairment were present in substantial proportions, unidentified by routine assessment of older patients. PMID- 15881560 TI - Iodine prophylaxis, goitre and thyroid autoimmunity in Sri Lanka. PMID- 15881561 TI - Disaster management: lessons from immediate responses to the tsunami. AB - There are many lessons learnt from the immediate reactions of people in the aftermath of the vast destruction from the giant waves that hit Sri Lanka. Reactions of victims fell between extremes of resilience and helplessness. Responses of those not directly harmed illustrated the two extremes--selflessness and depravity. These responses offer insight into how we live and how we react to situations, and also to an extent how we should be reacting. Among these is that helpers should respect and involve, from the inception, the resources within the group or community affected by the disaster. Control of the relief effort to the maximum feasible extent, should be in the hands of those at whom it is directed. PMID- 15881562 TI - An unusual case of focal epilepsy. PMID- 15881563 TI - An unusual presentation of female urethral leiomyoma. AB - Leiomyoma of the urethra is a rare clinical entity. We report an unusual presentation of this tumour, which led to a diagnostic and surgical dilemma. The patient was a 16-year old female who presented with a labial mass which was palpable abdominally. Imaging methods and laparoscopy demonstrated a well defined soft tissue mass arising from the pelvis, without any obvious involvement of the urinary tract. The tumour (8 x 10 cm) was completely excised with reconstruction of the bladder and urethra. Histology confirmed a cellular leiomyoma. PMID- 15881564 TI - Distal aphalangia, microcephaly and mental retardation. PMID- 15881565 TI - Anaphylactic shock and acute myocardial infarction following intravenous ceftazidime. PMID- 15881566 TI - Is there a delay in diagnosing tuberculosis? Experience in a Teaching Hospital. PMID- 15881567 TI - Use and interpretation of phrases in histopathology reports. PMID- 15881568 TI - Single carbon nanotube optical spectroscopy. AB - This Minireview discusses novel insights into the electronic structure of carbon nanotubes obtained using single-molecule fluorescence spectroscopy. Fluorescence spectra from single nanotubes are well described by a single, Lorentzian lineshape. Nanotubes with identical structures fluoresce with different energies due to local electronic perturbations. Carbon nanotube fluorescence unexpectedly does not-show any intensity or spectral fluctuations at 300 K The lack of intensity blinking or bleaching demonstrates that carbon nanotubes have the potential to provide a stable, single-molecule infrared photon source, allowing for the exciting possibility of applications in quantum optics and biophotonics. PMID- 15881569 TI - Double-tunnel nanoscale switch with a redox mediator: operational principles and tunneling spectroscopy. AB - A description of the physical mechanism and operation of a novel nanometric electronic switch [D.I. Gittins et al., Nature 2000 408, 67] is presented. New options for controlling the properties of this device are suggested and analyzed. PMID- 15881570 TI - Synergistic enhancement of the solubility of hexamethylenetetramine in subcritical CO2-ethanol mixtures studied by infrared spectroscopy. PMID- 15881571 TI - Multiwalled carbon nanotubes covalently modified with fast black K. PMID- 15881572 TI - Towards Raman optical activity calculations of large molecules. PMID- 15881573 TI - Engineering the electronic structure of single-walled carbon nanotubes by chemical functionalization. PMID- 15881574 TI - Photochemical alpha-cleavage in pyran-2-thione: generation of aldehydethioketene and thioaldehydeketene photoproducts. PMID- 15881575 TI - Direct NMR evidence for the presence of mobile surface hydrides on ruthenium nanoparticles. PMID- 15881576 TI - On differences between hydrogen bonding and improper blue-shifting hydrogen bonding. AB - Twenty two hydrogen-bonded and improper blue-shifting hydrogen-bonded complexes were studied by means of the HF, MP2 and B3LYP methods using the 6-31G(d,p) and 6 -311 ++G(d,p) basis sets. In contrast to the standard H bonding, the origin of the improper blue-shifting H bonding is still not fully understood. Contrary to a frequently presented idea, the electric field of the proton acceptor cannot solely explain the different behavior of the H-bonded and improper blue-shifting H-bonded complexes. Compression of the hydrogen bond due to different attractive forces-dispersion or electrostatics--makes an important contribution as well. The symmetry-adapted perturbation theory (SAPT) has been utilized to decompose the total interaction energy into physically meaningful contributions. In the red shifting complexes, the induction energy is mostly larger than the dispersion energy while, in the case of blue-shifting complexes, the situation is opposite. Dispersion as an attractive force increases the blue shift in the blue-shifting complexes as it compresses the H bond and, therefore, it increases the Pauli repulsion. On the other hand, dispersion in the red-shifting complexes increases their red shift. PMID- 15881577 TI - Noncovalent complexes between dimethyl ether and formic acid--an ab initio and matrix isolation study. AB - The complexes formed by noncovalent interactions between formic acid and dimethyl ether are investigated by ab initio methods and characterized by matrix isolation spectroscopy. Six complexes with binding energies between -2.26 and -7.97 kcal mol(-1) (MP2/cc-pVTZ+zero point vibrational energy+basis set superposition erros) are identified. The two strongest bound complexes are, within a range of 0.3 kcal mol(-1), isoenergetic. The binding in these six dimers can be described in terms of OH...O, C=O...H, C-O...H and CH...O interactions. Matrix isolation spectroscopy allowed to characterize the two strongest bound complexes by their infrared spectra. PMID- 15881578 TI - Solvent-dependent photoacidity state of pyranine monitored by transient mid infrared spectroscopy. AB - We investigate with femtosecond mid-infrared spectroscopy the vibrational-mode characteristics of the electronic states involved in the excited-state dynamics of pyranine (HPTS) that ultimately lead to efficient proton (deuteron) transfer in H2O (D2O). We also study the methoxy derivative of pyranine (MPTS), which is similar in electronic structure but does not have the photoacidity property. We compare the observed vibrational band patterns of MPTS and HPTS after electronic excitation in the solvents: deuterated dimethylsulfoxide, deuterated methanol and H2O/D2O, from which we conclude that for MPTS and HPTS photoacids the first excited singlet state appears to have charge-transfer (CT) properties in water within our time resolution (150 fs), whereas in aprotic dimethylsulfoxide the photoacid appears to be in a non-polar electronic excited state, and in methanol (less polar and less acidic than water) the behaviour is intermediate between these two extremes. For the fingerprint vibrations we do not observe dynamics on a time scale of a few picoseconds, and with our results obtained on the O-H stretching vibration we argue that the dynamic behaviour observed in previous UV/Vis pump-probe studies is likely to be related to solvation dynamics. PMID- 15881579 TI - Autoxidation of cyclohexane: conventional views challenged by theory and experiment. AB - In spite of its industrial importance, the detailed reaction mechanism of cyclohexane autoxidation by O2 is still insufficiently known. Based on quantum chemical potential energy surfaces, rate coefficients of the primary and secondary chain propagation steps involving the cyclohexylperoxyl (CyOO) radical were evaluated using multiconformer transition-state theory. Including tunneling and hindered-internal-rotation effects, the rate coefficient for hydrogen-atom abstraction from cyclohexane (CyH) by CyOO was calculated to be k(T)= 1.46 x 10( 11) x exp(-17.8 kcal mol(-1)/ RT) cm3s(-1) (300-600K), close to the experimental data. A "Franck-Rabinowitch cage" reaction between the nascent cyclohexylhydroperoxide (CyOOH) and cyclohexyl radical, products from CyOO + CyH, is put forward as an initially important cyclohexanol (CyOH) formation channel. alphaH abstraction by CyOO. from cyclohexanone was calculated to be only about five times faster than that from CyH, too slow to explain all the observed side products. The a-hydrogen (alphaH) abstractions from CyOH and CyOOH by CyOO. are predicted to be about 10 and 40 times faster, respectively, than the CyOO. +CyH reaction. The very fast CyOO.+CyOOH reaction proceeds through the unstable Cy alphaH .OOH radical that decomposes spontaneously into the ketone (Q=O) plus the OH radical; the "hot" .OH is found to produce the bulk of the alcohol via a second, "activated cage" reaction analogous to that above. It is thus shown how the very reactive CyOOH intermediate is the predominant source of ketone and alcohol, while it also leads to some side products. The alpha hydroxycyclohexylperoxyl radical formed during the moderately fast oxidation of CyOH is shown to decompose fast into HO2 + cyclohexanone in a rapidly equilibrated reaction, which constitutes a smaller, second ketone source. These two fast cyclohexanone forming routes avoid the need for unfavorable molecular routes hitherto invoked as ketone sources. The theoretical predictions are supported and complemented by experimental findings. The newly proposed scheme is also largely applicable to the oxidation of other hydrocarbons, such as toluene, xylene, and ethylbenzene. PMID- 15881580 TI - Kinetics and stability of CH4-CO2 mixed gas hydrates during formation and long term storage. AB - The formation of CH4-CO2 mixed gas hydrates was observed by measuring the change of vapor-phase composition using gas chromatography and Raman spectroscopy. Preferential consumption of carbon dioxide molecules was found during hydrate formation, which agreed well with thermodynamic calculations. Both Raman spectroscopic analysis and the thermodynamic calculation indicated that the kinetics of this mixed gas hydrate system was controlled by the competition of both molecules to be enclathrated into the hydrate cages. However, the methane molecules were preferentially crystallized in the early stages of hydrate formation when the initial methane concentration was much less than that of carbon dioxide. According to the Roman spectra, pure methane hydrates first formed under this condition. This unique phenomenon suggested that methane molecules play important roles in the hydrate formation process. These mixed gas hydrates were stored at atmospheric pressure and 190 K for over two months to examine the stability of the encaged gases. During storage, CO2 was preferentially released. According to our thermodynamic analysis, this CO2 release was due to the instability of CO2 in the hydrate structure under the storage conditions. PMID- 15881581 TI - Dynamic evolution of Kohn-Sham electron density in the real-time domain with finite basis expansion. AB - We present a novel method for time-dependent density functional theory calculations on dynamic linear response and electron density evolution in the real-time domain with the finite basis expansion approach of conventional quantum chemistry. To demonstrate the validity and efficiency of this method, dynamic polarizabilities of a water chain and diphenylene molecules are computed by employing the Chebyshev interpolation algorithm, which was developed by Baer and co-workers. The calculated dynamic polarizabilities show good agreement with those obtained from conventional linear response calculations. The density evolution in the real-time domain with application of a long-duration electric field gives electronic conduction in molecules, where a dynamic process of charge transfer is observed with the snapshots of response density in real time. Charge transfer oscillating with the frequency of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) gap is shown in a diphenylene molecule while there is little change in time for a water chain. PMID- 15881582 TI - Mechanical response analysis and power generation by single-cell stretching. AB - To harvest useful information about cell response due to mechanical perturbations under physiological conditions, a cantilever-based technique was designed, which allowed precise application of arbitrary forces or deformation histories on a single cell in vitro. Essential requirements for these investigations are a mechanism for applying an automated cell force and an induced-deformation detection system based on fiber-optical force sensing and closed loop control. The required mechanical stability of the setup can persist for several hours since mechanical drifts due to thermal gradients can be eliminated sufficiently (these gradients are caused by local heating of the cell observation chamber to 37 degrees C). During mechanical characterization, the cell is visualized with an optical microscope, which enables the simultaneous observation of cell shape and intracellular morphological changes. Either the cell elongation is observed as a reaction against a constant load or the cell force is measured as a response to constant deformation. Passive viscoelastic deformation and active cell response can be discriminated. The active power generated during contraction is in the range of Pmax= 10(-16) Watts, which corresponds to 2500 ATP molecules s(-1) at 10 k(B)T/molecule. The ratio of contractive to dissipative power is estimated to be in the range of 10(-2). The highest forces supported by the cell suggest that about 10(4) molecular motors must be involved in contraction. This indicates an energy-conversion efficiency of approximately 0.5. Our findings propose that, in addition to the recruitment of cell-contractile elements upon mechanical stimulation, the cell cytoskeleton becomes increasingly crosslinked in response to a mechanical pull. Quantitative stress-strain data, such as those presented here, may be employed to test physical models that describe cellular responses to mechanical stimuli. PMID- 15881583 TI - Theoretical study of inversion and topomerization processes of substituted cyclohexanes: the relevance of the energy 3D hypersurface. AB - Although the potential energy surface of highly symmetric cyclohexane has been extensively reviewed, no attention has been paid to the study of the effect of substitution of a methylene group by a heteroatom. The substitution may cause changes in molecular symmetry as well as the dipole moment, and the unshared electron pairs associated with the heteroatom may also introduce changes in molecular reactivity. However, these phenomena are not yet completely understood. To address these issues, a rigorous description of the inversion-topomerization process of methylcyclohexane and a revision of the conformational potential energy of oxane and thiane are presented. Moreover, the usefulness of providing a three-dimensional representation of these processes is discussed. In the case of methylcyclohexane, calculations show that three transition states are associated with inversion and four more with topomerization. In contrast, for oxane and thiane, only two transition states are involved with inversion and two with topomerization. Two fundamental conclusions can be drawn from this study. The first is that the inversion process occurs through elementary, stages that we have denoted "conformational elemental stages", which is an analogous term to that used for reaction mechanism description (minima-transition state-minima) where several elemental steps take place. The second conclusion is that two independent processes, inversion and topomerization, are connected by some common conformers. The inversion process controls the ring interchange, while topomerization allows exchange between skewed boats. PMID- 15881584 TI - Conformational and electronic properties of a microperoxidase in aqueous solution: a computational study. AB - A theoretical study of the conformational properties of a small heme peptide in aqueous solution is carried out by classical, long-timescale molecular dynamics simulations. The electronic properties of this species, that is, the relative energies of its excited electronic states and the redox potential, are reproduced and related to the conformational behavior using the perturbed matrix method and basic statistical mechanics. Our results show an interesting coupling between the conformational transitions and the electronic properties. These investigations, beyond the biophysically relevant results addressing the long-standing question of the actual role of the enzyme structure on the enzyme activity, are also of some methodological interest since they offer a further computational perspective for including the electronic degrees of freedom into the modeling of rather complex molecular systems. PMID- 15881585 TI - A kinetic model to describe nanocrystal growth by the oriented attachment mechanism. AB - The classical model of particle coagulation on colloids is revisited to evaluate its applicability on the oriented attachment of nanoparticles. The proposed model describes well the growth behavior of dispersed nanoparticles during the initial stages of nanoparticle synthesis and during growth induced by hydrothermal treatments. Moreover, a general model, which combines coarsening (i.e., Ostwald ripening) and oriented attachment effects, is proposed as an alternative to explain deviations between experimental results and existing theoretical models. PMID- 15881586 TI - The sound velocity in ideal liquid mixtures from thermal volume fluctuations. AB - The selection of the correct mixing rule for sound velocity in ideal liquid mixtures determines the interpretation of the sound velocity in real mixtures. This is especially important for the determination of apparent properties of solutes, such as their apparent compressibility. There are different approaches reported in the literature, and this article presents a new derivation of the mixing rule based on statistical mechanics. It is shown that the correlation of volume fluctuations between adjacent components has a crucial influence on the ideal mixing rule. PMID- 15881587 TI - Possible pathways of CVD processes leading to III-V semiconductors via a two dimensional growth. AB - Quantum-chemical calculations were carried out to shed more light upon possible first intermediates formed during chemical vapour deposition (CVD) processes leading to III-V semiconductors. Information about possible structures of intermediates and about thermodynamic properties in dependence of the temperature were collected. Because some of the systems calculated herein contain a substantial number of atoms (up to 144), it is limited to intermediates on the way to solid AIN. According to our suggestion, the first intermediates are species with the overall formula AINH2, Al2N2H4, Al3N3H6, AI5N5H8, AI7N7H, and Al9N9H, AI12N12H12. In further calculations the growth of a second layer, on top of the first layer, which is modelled by the Al12N12H12 cluster, is simulated. The Al12N12H12 "surface" offers two places for the attachment of an Al3N3H6 ring. At temperatures of 1000 K, the deltaG value for this attachment is only slightly negative, which suggests that the molecules can move relatively freely on the surface. This might be of importance for a defect-free growth process. Up to four layers are built on top of the first Al12N12H12 layer leading to Al24N24H24 (two layers), Al36N36H36 (three layers) and Al48N48H48 (four layers). The structures are compared with that of solid AIN in its most stable Wurzit-type structure. PMID- 15881588 TI - Bandgap widening of titania through semiconductor support interactions. AB - Silica-supported titania powders with 50, 36, 13 and 4 wt% of TiO2 (TiO2-50/SiO2, TiO2-36/SiO2, TiO2-13/SiO2 and TiO2-4/SiO2) were prepared by hydrolysis of TiCl4 in the presence of silica, followed by calcination at 500 degrees C. The formation of Ti-O-Si linkages was confirmed by diffuse reflectance infrared Fourier transform spectroscopy. Atomic force microscopy indicated the presence of titania crystals larger than 15 nm. All supported materials exhibited a blue shift of the TiO2 absorption edge, which was attributed to an electronic semiconductor support interaction. Bandgap energies of TiO2-50/SiO2, TiO2 36/SiO2, TiO2-13/SiO2 and TiO(2)4/SiO2 were measured to be 3.28, 3.36, 3.40 and 3.42 eV, respectively, as compared to 3.15 eV for unsupported TiO2. From these values, and from the quasi-Fermi level of electrons, a high anodic shift of both the valence and the conduction band was estimated. X-ray photoelectron spectroscopy (XPS) measurements of oxygen 1s- and titanium 2p-binding energies confirmed the anodic shift of the band edges. PMID- 15881589 TI - Ab initio calculations and Franck-Condon simulation of the absorption spectra of GeCl2 including anharmonicity. AB - Geometrical parameters, vibrational frequencies and relative electronic energies of the X1A1, a3B1 and A1B1 states of GeCl2 have been calculated at the CCSD(T) and/or CASSCF/MRCI level with basis sets of up to aug-cc-pV5Z quality. Core electron correlation and relativistic contributions were also investigated. RCCSD(T)/ aug-cc-pVQZ potential energy functions (PEFs) of the X1A1 and a3B, states, and a CASSCF/MRCl/aug-cc-pVQZ PEF of the A1B1 state of GeCl2 are reported. Anharmonic vibrational wavefunctions of these electronic states of GeCl2, obtained variationally using the computed PEFs, are employed to calculate the Franck-Condon factors (FCFs) of the a-X and A-X transitions of GeCl2. Simulated absorption spectra of these transitions based on the computed FCFs are compared with the corresponding experimental laser-induced fluorescence (LIF) spectra of Karolczak et al. [J. Chem. Phys. 1993, 98, 60-70]. Excellent agreement is obtained between the simulated absorption spectrum and observed LIF spectrum of the a-X transition of GeCl2, which confirms the molecular carrier, the electronic states involved and the vibrational assignments of the LIF spectrum. However, comparison between the simulated absorption spectrum and experimental LIF spectrum of the A-X transition of GeCl2 leads to a revision of vibrational assignments of the LIF spectrum and suggests that the X1A1 state of GeCl2 was prepared in the experimental work, with a non-Boltzmann vibrational population distribution. The X(0,0,1) level is populated over 4000 times more than expected from a Boltzmann distribution at 60 K, which is appropriate for the relative population of the other low-lying vibrational levels, such as the X(1,0,0) and X(0,1,0) levels. PMID- 15881590 TI - Photophysical and electrochemical properties of meso, meso-linked oligoporphyrin rods with appended fullerene terminals. AB - The electrochemical and photophysical properties of molecular architectures consisting of oligomeric meso,meso-linked oligoporphyrin rods linked at both extremities to methanofullerene moieties are presented in comparison to those of model systems. Cyclic voltammetry data evidence the presence of a strong intramolecular electronic coupling along the porphyrin oligomers that varies slightly with their length. This interaction affects the redox potentials of both fullerene and porphyrin moieties. The electronic coupling between the two chromophores is confirmed by comparing the redox potentials of porphyrin arrays before and after attachment of the carbon sphere. Electronic absorption, fluorescence, and phosphorescence spectra of the porphyrin oligomers in toluene are reported, which provide the energy of the lowest singlet and triplet electronic excited states. In the fullerene-porphyrin conjugates, ground-state charge-transfer (CT) interactions are evidenced by low-energy absorption features above 750 nm. These systems also exhibit near-infrared (NIR) CT luminescence in toluene with lifetimes shorter than 1000 ps. On increasing the solvent polarity (from toluene to Et2O and THF), CT emissions become progressively weaker, red shifted, and shorter lived, which reflects the energy-gap law and Marcus inverted region effects. Luminescence is not detected in benzonitrile. Picosecond transient absorption spectroscopy of the porphyrin-fullerene conjugates allows detection of the porphyrin cation as a clear fingerprint for electron transfer. The rate of charge recombination is in agreement with CT luminescence lifetimes, which confirms the occurrence of NIR radiative back-electron transfer. PMID- 15881591 TI - Influence of pH on the appearance of active peptides in the course of peptic hydrolysis of bovine haemoglobin. AB - Influence of pH on the appearance of active peptides in peptic hydrolysis of bovine haemoglobin was studied in a homogenous phase system. Six active peptides were studied: three hemorphins: LVVH-7 (beta 31-40), VVH-7 (beta 32-40), VVH-4 (beta 32-37), one bradykinin-potentiating peptide (alpha 110-125), one antibacterial peptide (alpha 1-23), and neokyotorphin (alpha 137-141). The influence of pH was investigated in the course of the hydrolysis of haemoglobin by pepsin at 23 degrees C in acetate buffer at pH 3.5, pH 4.5, and pH 5.5. The hydrolysis of haemoglobin was studied in the presence or absence of urea. The haemoglobin hydrolysis at pH 4.5 is taken as a reference. Two different mechanisms of hydrolysis were observed: "one by one" for native haemoglobin hydrolysis at pH 4.5 and 5.5, and "zipper" for denatured haemoglobin at pH 3.5, pH 4.5, and pH 5.5, and native haemoglobin at pH 3.5. Whatever the pH and medium, a selectivity change by the pepsin was noticed. In the presence of urea, there are two phenomena: some peptides are preferentially produced at pH 3.5 and other peptides at pH 5.5, which seems to favour one particular site of pepsin that is cut. In the absence of urea, these active peptides reached a higher concentration at pH 3.5. In order to prepare these six active peptides, it is suitable to hydrolyse haemoglobin in the absence of urea at pH 3.5 (this pH denatures haemoglobin) where a "zipper" mechanism is obtained, and the peptide quantity is more significant at pH 3.5 than at pH 4.5. PMID- 15881592 TI - Immobilization of alpha-glucosidase in chitosan coated polygalacturonic acid. AB - Crude alpha-glucosidase from Baker's yeast was immobilized in polygalacturonic acid beads and coated with chitosan. Chemical and physical characterization were performed by using p-nitrophenyl-alpha-D-glucopyranoside (pNPG) as an artificial substrate. Operation, thermal, pH, and strorage stabilities of the free and immobilized enzyme were also examined. The stabilities of immobilized enzyme were found to be better than that of the free enzyme. Furthermore, the hydrolysis rate of the chitosan coated alpha-glucosidase polygalacturonic acid beads were studied. In conclusion, the enzyme beads appear to have good characteristics and offer the prospect that this system may find application in enzyme immobilization, in addition to controlled drug release studies. PMID- 15881593 TI - Synthesis of mixed esters of ascorbic acid using methyl esters of palm and soybean oils. AB - Mixed esters of ascorbic acid were synthesized using methyl esters of palm and soybean oils as acyl donors, in acetone at 50 degrees C, and catalyzed by Novozym 435. A conversion of 62% was obtained with palm oil methyl ester at an ascorbic acid to acyl donor molar ratio of 1:4; the mixed ester contained 45.89% ascorbyl palmitate, 42.59% ascorbyl oleate and 10.1% ascorbyl linoleate. Acylation with soybean oil methyl ester resulted in 17% conversion, yielding a mixed ester containing 10.08% ascorbyl palmitate, 20.68% ascorbyl oleate, and 64.96% of ascorbyl linoleate. The mixed esters of ascorbic acid can find direct use in food and cosmetics. PMID- 15881594 TI - Recombinant NY-ESO-1 cancer antigen: production and purification under cGMP conditions. AB - The cancer-testis antigen, NY-ESO-1, has been engineered into a bacterial expression plasmid which incorporates a His6-tag. The plasmid was transfected into E. coli strain BL21 and Master and Working cell banks generated from this expression system. Three 15-litre fermentations were performed under cGMP (code of Good Manufacturing Practice) conditions and the crude NY-ESO-1 tagged protein isolated as solubilised inclusion bodies. A three-step cGMP chromatography process (immobilised metal affinity, anion exchange, and hydrophobic interaction) was utilised to purify the protein. The purified NY-ESO-1 is being used in early stage human cancer vaccine trials in Australia and the U.S.A. PMID- 15881595 TI - Preparation and characterization of kappa-carrageenan immobilized urease. AB - Urease was encapsulated within kappa-carrageenan beads. Various parameters, such as amount of kappa-carrageenan and enzyme activity, were optimized for the immobilization of urease. Immobilized urease was thoroughly characterized for pH, temperature, and storage stabilities and these properties were compared with the free enzyme. The free urease activity quickly decreased and the half time of the activity decay was about 3 days at 4 degrees C. The immobilized urease remained very active over a long period of time and this enzyme lost about 70.43% of its orginal activity over the period of 26 days for storage at 4 degrees C. The Michaelis constant (Km) and maximum reaction velocity (Vmax) were calculated from Lineweaver-Burk plots for both free and immobilized enzyme systems. Vmax = 227.3 U/mg protein, Km = 65.6 mM for free urease and Vmax = 153.9 U/mg protein, Km = 96.42 mM for immobilized urease showed a moderate decrease of enzyme specific activity and change of substrate affinity. PMID- 15881596 TI - Expression and purification of a Tuber borchii fruitbody-specific protein, TBF-1, from Escherichia coli: generation of polyclonal antibodies. AB - TBF-1 is a fruitbody-specific protein present in the white truffle species Tuber borchii Vittad. A similar protein has been found only in the closely related species Tuber dryophilum (TDF-1), but not in other truffles. The protein from T. borchii was overexpressed as fusion protein in E. coli and was purified to homogeneity by affinity chromatography. Recombinant protein was used for generating polyclonal antibodies. The antiserum strongly reacted with TBF-1, weakly recognized TDF-1, and did not detect correlate band in the other white truffle species. The high level of expression of this protein in the fruitbody and the specificity of the antibody anti-TBF-1 make it possible to set up a diagnostic tool for detecting these species in natural samples and foodstuffs. PMID- 15881597 TI - An efficient method for rapid amplification of Arisaema heterophyllum agglutinin gene using a genomic walking technique. AB - The genomic sequence of Arisaema heterophyllum agglutinin (AHA), a mannose binding lectin (MBL), was cloned through a novel genomic walking technique. Adaptor ligation reactions and subsequent amplifications with adaptor primer and multiple specific primers were used to generate specificity in this method. The method allowed for the amplification of over 1 kb of genomic DNA sequence immediately upstream and downstream from the 5' and 3' ends of full-length cDNAs. For aha gene, the upstream regions contained a putative transcription initiation start site and other sequences commonly found in eukaryotic promoters. The downstream regions of aha contained two polyadenylation signals. Our study demonstrated that aha had no intron like mannose-binding lectin genes cloned from other plant species so far. This efficient method, based on a genomic walking technique, was useful for the cloning of promoters, insertion sites, and other sequences of interest without constructing and screening genomic libraries. PMID- 15881598 TI - Production and properties of alpha-amylase from Penicillium chrysogenum and its application in starch hydrolysis. AB - Fungi were screened for their ability to produce alpha-amylase by a plate culture method. Penicillium chrysogenum showed high enzymatic activity. Alpha-amylase production by P. chrysogenum cultivated in liquid media containing maltose (2%) reached its maximum at 6-8 days, at 30 degrees C, with a level of 155 U ml(-1). Some general properties of the enzyme were investigated. The optimum reaction pH and temperature were 5.0 and 30-40 degrees C, respectively. The enzyme was stable at a pH range from 5.0-6.0 and at 30 degrees C for 20 min and the enzyme's 92.1% activity's was retained at 40 degrees C for 20 min without substrate. Hydrolysis products of the enzyme were maltose, unidefined oligosaccharides, and a trace amount of glucose. Alpha-amylase of P. chrysogenum hydrolysed starches from different sources. The best hydrolysis was determined (98.69%) in soluble starch for 15 minute at 30 degrees C. PMID- 15881599 TI - Enhancements and justice: problems in determining the requirements of justice in a genetically transformed society. AB - There is a concern that genetic engineering will exacerbate existing social divisions and inequalities, especially if only the wealthy can afford genetic enhancements. Accordingly, many argue that justice requires the imposition of constraints on genetic engineering. However, it would be unwise to decide at this time what limits should be imposed in the future. Decision makers currently lack both the theoretical tools and the factual foundation for making sound judgments about the requirements of justice in a genetically transformed society. Moreover, focusing on the uncertain inequities of the future may result in failure to give priority to more pressing inequities of the present. Especially in a country that recently has enacted tax legislation that will widen existing wealth disparities, concern about the distant threat of a genetic aristocracy appears misplaced. PMID- 15881600 TI - Problems in the interpretation of nonaqueous titrations. AB - Most phenomena (like promoting and leveling as well as differentiating effects) of common nonaqueous titrations can be explained by the Bronsted's concept combined with the hydrogen bond theory but some open problems are shown in connection with the use of formic acid and/or acetic anhydride. PMID- 15881601 TI - Development of models for cytochrome P450 2A5 as well as two of its mutants. AB - It is known that small changes in the amino acid sequence can change the catalytic activity of cytochromes towards substrates dramatically. With the aim to broaden our knowledge about the structural properties of cytochromes and their relation with substrate specificity a model of CYP2A5 was built by homology modelling based on the crystal structure of CYP2C5. Model stability was evaluated by subjection of the model to a free molecular dynamics simulation in a waterbox under almost physiological conditions using the GROMACS program. The protein folding remains stable over 1.5 ns under these conditions. The modelling procedure was repeated for two mutated forms of CYP2A5 with known differing substrate selectivities towards corticosterone and desoxycorticosterone. A detailed analysis of the models and their behaviour in long running molecular dynamics simulations allows an understanding of the requirements for enzyme activity as well as an explanation of respective experimental data on the molecular level. PMID- 15881602 TI - Quantitative structure activity relationship studies of diaryl thiophen derivatives as selective COX-2 inhibitors. AB - QSAR studies were carried out on a series of diaryl thiophens that act as selective COX-2 inhibitors using Molecular Operating Environment (MOE). The studies were carried out on a training set of 20 analogs. These studies produced good predictive models and indicated that partial Charge Descriptors (Q_VSA FPNEG), shape descriptor (STD_DIM 3); hydrophobic descriptor (LOG P) and electronic descriptor (TPSA-Topological positive surface area) contribute to selective cyclooxygenase-2 inhibitory activity. Pharmacophoric query generation showedthe pharmacophores required for selective COX-2 inhibitory activity. PMID- 15881603 TI - [12-acyl-6,1 2-dihydro-6-phenyl-[1 ]benzofuro[2,3-c]- and -[1,5]benzothieno[2,3 c]-[1 ,5]-benzothiazepines--tetracyclic diltiazem derivatives]. AB - The reaction of the tetracycles 1 with chloroacetylchloride yields the amides 2. The structure of 2a was proven by X-ray analysis. The amides 2 exist as diastereomers in solution because of planar chirality. From the N-chloroacetyl compounds only 2a, b could be substituted with diethylamine to give 3a, b. Reduction experiments of 3a, b with DIBAH do not afford diltiazem analogues; instead, the starting compounds la, b are formed by hydrolysis. PMID- 15881604 TI - Development and validation of an UV spectrophotometric method for determination of gatifloxacin in tablets. AB - A simple, sensitive and accurate spectrophotometric method was developed for the assay of gatifloxacin in raw material and tablets. Validation of the method yielded good results concerning range, linearity, precision and accuracy. The absorbance was measured at 287 nm for gatifloxacin tablet solutions. The linearity range was found to be 4.0-14.0 microg/mL for gatifloxacin. It was also found that the excipients in the commercial tablets did not interfere with the method. PMID- 15881605 TI - Spectrofluorometric determination of some beta-blockers in tablets and human plasma using 9,10-dimethoxyanthracene-2-sodium sulfonate. AB - A simple and sensitive spectrofluorometric method was developed for the quantitative determination of some beta-blockers, namely arotinolol, atenolol and labetalol as hydrochloride salts. The method is based on the reaction of these drugs as n-electron donors with the fluorogenic reagent 9,10-dimethoxy-2 anthracene sulfonate (DMAS) as pi-acceptor in acidic medium. The obtained ion pairs were extracted into chloroform and measured spectrofluorometrically at 452 nm after excitation at 385 nm. The fluorescence intensity-concentration plots are rectilinear over the ranges of 0.5-5 microg x ml(1), 1.0-11.0 microg x ml(1) and 0.6-6.4 microg x ml(1) for labetalol, atenolol and arotinolol, respectively. The different parameters affecting the reaction pathway were thoroughly studied and optimized. No interference was observed from the common pharmaceutical excipients. The proposed method was successfully applied to the analysis of tablets and the results were statistically compared with those obtained by reference methods. The method was further extended to the in vitro determination of the drugs in spiked human plasma, the % recoveries (n = 3) ranged from 96.98 +/- 1.55 to 98.28 +/- 2.19. A proposal of the reaction pathway was postulated. PMID- 15881606 TI - [Color reaction of chlorhexidine and proguanil with hypobromite]. AB - Colour reaction of chlorhexidine and proguanil with hypobromite The antimalarial agent proguanil reacts with hypobromite to yield the red coloured (E)-3-[(4 chlorophenyl)imino]-N-isopropyl-3H-1,2,4-triazol-5-amine (6B). The structure of 6B was proven by X-ray. The red colour obtained by the test for the disinfectant chlorhexidinedihydrochloride Ph. Eur. with hypobromite is probably attributable to a corresponding chromophore. PMID- 15881607 TI - Investigations on 5-fluorouracil solid lipid nanoparticles (SLN) prepared by hot homogenization. AB - The purpose of this study was to investigate the effect of different formulation factors on the properties of solid lipid nanoparticles (SLN) prepared by a hot homogenization method. Using the particle size, physical stability constant (K(e)) and zeta-potential as standards, the stability of SLNs was investigated as a function of phospholipid and poloxamer contents. It was demonstrated that the content of phospholipid had a significant influence on the zeta-potential, which increased considerably with increasing phospholipid content. However, the particle size increased remarkably when the phospholipid content was as high as 1.5% due to the increased viscosity. Poloxamer 188 exhibited no remarkable influence on particle size when the concentration was as low as 1.0%. The influence of the phospholipid and poloxamer content on the embedding ratios of drug substances was further studied using 5-fluorouracil (5-Fu) as a model drug. It was shown that the embedding ratio increased considerably with phospholipid content and independent of poloxamer content, implying that 5-Fu was incorporated into the phospholipid bilayer membrane. PMID- 15881608 TI - Effect of processing variables on the release and particulate properties of sustained release amoxicillin microcapsules prepared by emulsion solvent evaporation. AB - This study reports the preparation of amoxicillin microcapsules by an emulsion solvent evaporation process. In particular the effect of processing variables including the dimension and position of stirring paddle and container; volume of continuous phase versus dispersion phase; stirring speed and encapsulation temperature on the release and particulate properties of the amoxicillin microcapsules were determined. When the diameter of the paddle was half of that of the container and the clearance between the paddle and the bottom of the vessel was 1/4 of the total volume in the vessel, almost no material stuck to the inside wall of the beaker and uniform microcapsules were prepared. Very uniform, round microcapsules were also prepared with a high yield when V(acetone): V(light mineral oil) = 1 : 3 and 1 : 5 because these systems ensured the formation of uniform emulsions. Physical evaluation of the microcapsules also showed that optimum drug release was achieved when the microcapsules were round, did not aggregate, were protected from the burst effect, the stirring speed for preparation was between 600-800 rpm and evaporation temperature was 25 degrees C. Microcapsules prepared using these ideal conditions achieved constant amoxicillin release for up to 12 h. PMID- 15881609 TI - Characterization of the force effect of aqueous and oily eye drops. AB - During the instillation of eye drops the drop impact affects a mechanical irritation on the eye surface. The force impact occurring in the moment of impact may be measured in vitro. Four commercially available eye drop preparations of different consistency and viscosity were tested. In dependance on the drip distance, the quantity of force impact was determined as well as the maximally effecting force during impact in each case. Whereas the force impact increased with all preparations with rising drip distance, only the higher viscosity solutions showed an approximate linear increase of the maximal force. With the aqueous low-viscosity eye drops, peak forces were ascertained. The drop oscillation was considered as the cause. The test results show that the consistency of eye drop preparations and their resulting physical-chemical properties like viscosity influence the quantity of the maximal force during the drop impact. PMID- 15881610 TI - Preparation and in vitro characterization of a semi-solid dispersion of flurbiprofen with Gelucire 44/14 and Labrasol. AB - Flurbiprofen is characterized by low solubility in water and has been implicated in causing gastro intestinal ulceration. The purpose of this study was to increase the dissolution characteristics of flurbiprofen by preparing a semi solid dispersion with Gelucire 44/14 and Labrasol (F1) in hard gelatin capsules. The results were evaluated by comparing several in vitro parameters with powdered drug filled into hard gelatin capsules. The in vitro dissolution testing of the dosage forms was performed in different media (simulated gastric fluid, pH 1.2; citrate buffer pH 4.5; phosphate buffers pH 6.8 and 7.2, and water). Characterization of semi-solid dispersions and physical mixtures was performed using Fourier transform-infrared spectroscopy (FT-IR), Differential scanning calorimetry (DSC), particle size analysis and turbidity measurement. The results suggest that all semi-solid dispersions of flurbiprofen showed a remarkable improvement in the rate and extent of drug dissolution. The dissolution of F1 exhibited significant improvement in all dissolution media at different pH. The dissolution of flurbiprofen within 30 min in pH 1.2 was (55%), in pH 4.5 67%, pH 6.8 96%, pH 7.2 98% and in water 88%. FT-IR indicated no strong drug: excipient interactions, and DSC studies indicated a loss of crystalline nature of the drug. The particle size analysis revealed an average size diameter from 194 to 278 nm. Therefore, a semi-solid dispersion of flurbiprofen with Gelucire and Labrasol may have the potential of improved bioavailability because of the enhanced in vitro properties. PMID- 15881611 TI - Indirect biochemical evidence that reserpine methiodide produces selective depletion of peripheral biogenic amines in rats. AB - Reserpine, an alkaloid from Rauwolfia serpentina was widely used for its antihypertensive action in the past. In the present investigation, reserpine methiodide (RMI), a quaternary analogue of reserpine was synthesised and evaluated biochemically for its central and peripheral amine depleting actions in rats and compared with reserpine. The 24 h urinary excretion of vanillylmandelic acid (VMA), 5-hydroxyindoleacetic acid (5-HIAA) and homovanillic acid (HVA), the respective metabolites of noradrenaline, serotonin and dopamine were estimated and considered as indirect biochemical indices for the amine depleting action of reserpine and RMI. The results indicate that RMI at doses of equal to and double the equimolar doses of reserpine was found to deplete the peripheral amines without affecting the central stores of the amines. The results further suggest that the quaternization of reserpine might restrict its transfer across the blood brain barrier and could be the reason for its selective peripheral action. PMID- 15881612 TI - Sphingosine 1-phosphate is involved in cytoprotective actions of calcitriol in human fibroblasts and enhances the intracellular Bcl-2/Bax rheostat. AB - Calcitriol is originally known to decrease proliferation rates of several carcinoma cells, partly via induction of apoptosis. On the other hand, the secosteroid is revealed to protect some cell types like thyrocytes, HL-60 cells and melanocytes against programmed cell death. Here we report that calcitriol despite its strong antiproliferative effect on human dermal fibroblasts did not induce apoptosis in these cells. In contrast, calcitriol possessed an antiapoptotic action in dermal fibroblasts. Thus, the ability of the apoptotic stimuli TNFalpha/actinomycin and C2-ceramides (C2-Cer) to induce programmed cell death was drastically diminished in the presence of calcitriol. Moreover, we identified sphingosine 1-phosphate (S1P) as a downstream mediator of calcitriol for its cytoprotective property. Thus, the secosteroid could not protect fibroblasts from apoptosis in the presence of N,N-dimethylsphingosine (DMS), which inhibits sphingosine kinase, the crucial enzyme to form S1P. Like calcitriol, S1P in different concentrations did not induce fibroblast apoptosis and moreover drastically decreased the rates of apoptotic cells after treatment with TNFalpha1/actinomycin. As S1P has been identified to modify the Bcl-2/ Bax ratio in epithelial cells and keratinocytes, we also measured the expression of these proteins in dermal fibroblasts revealing an increased Bcl-2 level after stimulation with S1P while the Bax protein expression was not modified. In conclusion, calcitriol H was revealed to protect human fibroblasts from apoptosis by formation of S1P resulting in a changed Bcl-2/Bax ratio. PMID- 15881613 TI - Biological activity of selected tyrosine-containing 2,5-diketopiperazines. AB - The study investigates two cyclic dipeptides, cyclo(Tyr-Tyr) (cTT) and cyclo(Phe Tyr) (cPT) with respect to their biological activity. Investigations using the whole-cell patch-clamp technique testing the effects of 100 microM cyclic dipeptide on ion channels, revealed reversible voltage-dependant blockade for cTT, while cPT exhibited irreversible time-dependant blockade of L-type calcium channels. The isolated retrogradely-perfused rat heart was used to determine the effects of 100 microM of either cTT or cPT on heart rate (HR), coronary flow rate (CFR), left ventricular systolic pressure (LVSP) and cardiac conduction speed. Results indicated opposing effects for the two compounds, where cTT increased HR and CF while cPT decreased HR, CF, LVSP as well as conduction speed. Other biological investigations included opioid binding and anti-neoplastic assays. Competitive binding curves, using tritiated DAMGO, revealed significant binding to micro-opioid receptors with IC50 values for cTT and cPT being 0.82 microM and 69.7 microM respectively. Anti-neoplastic activity was tested using three cultured cell lines: HT-29, MCF-7 and HeLa which were exposed to 2.5 mM cyclic dipeptide or 0.1 mM melphalan as a positive control. While cTT showed little activity against these lines, cPT resulted in as much as a 75.6% growth inhibition of MCF-7 cells, while also being active against HeLa (73.4% inhibition) and HT-29 (60.6%). The results indicate potential biological activity, showing a need for more investigation into tyrosine containing cyclic dipeptides and their analogues as potential bioactive compounds. PMID- 15881614 TI - Anti-inflammatory activity of extracts and a saponin isolated from Melilotus elegans. AB - The crude methanol extract of Melilotus elegans Ser. (Fabaceae), a plant widely used in Ethiopian traditional medicine for the treatment of asthma, haemorrhoid and lacerated wounds showed a significant anti-inflammatory activity against carrageenin-induced rat paw oedema. At a dose corresponding to 333.3 mg per kg body weight of dry plant material, the methanol extract displayed a strong inhibitory effect that was comparable to the inhibitory effect of 1 mg/kg of indomethacin in the same test system. Bioassay guided fractionation of the alcoholic extract led to the isolation of an oleanene-type triterpene saponin identified as azukisaponin V (1) ((3-O-[alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D glucopyranosyl(1 --> 2)-beta-D-glucuronopyranosyl]-soyasapogenol B). The structure of the compound was identified by using MS and extensive one- and two dimensional NMR experiments (1H, 13C, COSY, HMQC, HMBC and NOESY). One hour after injection of carrageenin, inhibition of oedema exerted by 1 was approximately ten times higher than that of indomethacin on a molar basis. PMID- 15881615 TI - Six new sesquiterpenes from Cacalia ainsliaeflora. AB - Five new eremophilane sesquiterpenes, 3beta,6beta-diangeloyloxy-8beta,10beta dihydroxyeremophilenolide (1); 6beta-acetoxy-3beta-angeloyloxy-8beta,10beta dihydroxyeremophilenolide (2); 3beta-angeloyloxy-6beta-methoxyeremophil 7(11),9(10)-dien-8alpha,12-olide (3), 3beta-angeloyloxy-8-oxo-eremophil-6(7)-en 12-oic acid (4); 3beta-angeloyloxy-10beta-hydroxy-8-oxo-eremophil-6(7)-en-12-oic acid (5), and a novel nor-eremophilane derivative, 3beta-angeloyloxy-10beta hydroxy-8-oxo-eremophil-6(7)-en (6), were isolated from the roots of Cacalia ainsliaeflora. Their structures were elucidated by spectroscopic methods, including 2DNMR. Compounds 1 and 2 were assayed against P388 and A549 Carcinoma cell lines. No positive activities were observed. PMID- 15881616 TI - Persistent suppression of phagocytosis after prolonged administration of local anesthetics carbisocaine and heptacaine in mouse leukocytes. AB - Effects of local anesthetics heptacaine and carbisocaine on mouse peritoneal macrophages after 5 days of peritoneal administration were observed. Both compounds in a daily dose 50 mg/kg caused significant decrease in phagocytic activity and index of phagocytosis (number of ingested particles). No significant effects on phagocytosis were observed when carbisocaine was administrated in a daily dose 5 mg/kg. No significant changes in weight gain and number of peritoneal macrophages were observed in all groups. PMID- 15881617 TI - Cytotoxic and antioxidant activity of Achillea alexandri-regis. AB - The cytotoxicity and antioxidant properties of herb extracts of Achillea alexandri-regis were studied. Combined chloroform and ethylacetate extracts exhibited a pronounced cytotoxic effect against HeLa cancer cells (IC50 = 25.92 +/- 4.96 microg/ml), and lower cytotoxicity against K562 leukemia cells (IC50 = 48.59 +/- 18.31 microg/ml). The methanol extract was found to be a moderately cytotoxic in vitro agent against HeLa and K562 cells. No suppressive activity was detected on non-malignant peripheral blood mononuclear cells (PBMC). The antioxidant activity of the methanol extract was assessed by DPPH radical scavenging. The methanol extract of A. alexandri-regis showed concentration dependent DPPH radical scavenging activity with IC50 = 36.14 +/- 0.05 microg/ml. PMID- 15881618 TI - [Transsexualism as an interdisciplinary phenomenon]. AB - The authors express their criticism on the currently prevailing Polish diagnostic and therapeutic criteria of transsexualism. Relying on their clinical experience and expertise (opinions for the court) as well as current literature, they point to the necessity of a discussion between specialists from various medical fields (psychiatry, sexology, urology, surgery, endocrinology, genetics) and humanistic sciences (psychology, sociology, law, ethics) on this interdisciplinary phenomenon. PMID- 15881619 TI - [Can psychiatry become neuropsychiatry?]. AB - Today more and more often there are prognoses that in the future psychiatry will have been absorbed by neurology. It would be thanks to the stormy progress of research on the neurophysiological, genetic and molecular foundations of mental disorders. The aim of the article is to assess the possibility as well as the supposed consequences of such an evolution of psychiatry. The considerations concern the peculiarity of the object of interest and the methods used in psychiatry in relation to the neurological object and methodology. This way the appraisal of raison d'etre of one common science: neuropsychiatry becomes possible. The question of fundamental importance for the evaluation of similarities and differences between the psychiatric and neurological perspectives is the way the psychophysical issue and especially the problem of the mind-brain relation are approached. The article presents the manners of solving these problems proposed by the contemporary philosophy of the mind. Together with parting with the full of errors and simplifications heritage of Descartes it appears the necessity to regard the presence of subjective mental states both conscious and unconscious in model of mind-brain relation. The example of such a solution is the biological naturalism of John Searle. The psychical life of the man in its subjective dimension remains the peculiar area of interests for psychiatry irrespective of the progress in research on the biological base of mental disorders. The especially valuable cognitive and therapeutic tool in this aspect is psychotherapy constituting the integral part of psychiatry. The present state of knowledge does not indicate that the psychotherapeutic wing of psychiatry can lose its importance and rather somewhat the contrary. The progress of neurobiology does not have to threaten the autonomy of psychiatry by any means and the maintenance of this autonomy depends decisively on the psychiatrists themselves. PMID- 15881620 TI - [Schizophrenia--neurodevelopmental and neurodegenerative theory revisited]. AB - The paper presents a critical review regarding the pathogenesis of schizophrenia. The neurodevelopmental theory which is now mostly recognized, needs some explanations, some elements of neurodegenerative theory are revisited. The developmental factors are confirmed as important in the pathogenesis of schizophrenia, but still it is unclear, how relatively subtle early damage causes so heavy thinking and emotional distortion like schizophrenia. There is also lack of evidence for neurodegeneration. Neuroimaging studies show the evidence for progression of brain abnormalities but at the same time there is no progression in neurocognitive disorders. Most likely late developmental disorders interfere with early brain damage, which leads to a neurocognitive disorder and clinical symptoms of schizophrenia. PMID- 15881621 TI - [Seasonality of birth in schizophrenia patients. Literature review]. AB - Schizophrenia is a major public health problem. The paper presents a review of the literature about seasonality of birth in schizophrenia. Tramer (1929) reported the first study on seasonality of birth in mental disorders. Seasonality of the birth of people who develop schizophrenia refers to a 5-8% excess of births in winter months. However, the peak of research into this phenomenon started in the 1990's. Torrey et al. (1997) reviewed over 250 studies from 29 countries in the Northern Hemisphere and 5 publications by authors from the Southern Hemisphere, concerning the relationship between birth seasonality and the incidence of schizophrenia. The research findings from the Northern Hemisphere were consistent, indicating a 5-8% increase in the risk for mental disorders in individuals born between December and May. Meta-analysis of investigations from the Northern Hemisphere indicates that seasonality of birth increases with geographical width. Results of research from other geographical areas, analysis of stability of this phenomenon during decades and attempts of explanation of seasonality are presented. PMID- 15881622 TI - [Could the fetus' exposure to influenza increase the risk of schizophrenia in adult life?]. AB - INTRODUCTION: Epidemiological studies indicate the existence of various environmental factors, which elevate the risk of schizophrenia in adult life. Influenza infection is a widely discussed causal factor in the relation to schizophrenia. The paper is a review of the studies on the theme of exposition to viral infections in foetal life--in connection with schizophrenia morbidity in adult life. Many of such studies, and yet not all of them show such a relationship. AIM: The aim of our study was to find the effect of influenza exposition in foetal life on the increase in schizophrenia morbidity of the Polish population, by applying the seasonal decomposition method. METHOD: All persons discharged from hospitals in Poland with a diagnosis of schizophrenia in the years 1997-2000 were taken up for the study. The data on the population births in the given months of the period 1964-1984 was also used. Monthly indices of the number of births of patients with a diagnosis of schizophrenia per 10,000 live births in the general population of men and women was applied in order to eliminate the seasonal variability due to the unstable number of births in the general population. The number of influenza cases in Poland was divided by the number of inhabitants and multiplied by 10,000 thus forming indices of influenza cases in the subsequent months of the period studied. The seasonal decomposition method--Census I--was used for the basic analyses--this being one of the methods of time interval analyses available in the Statistica programme. RESULTS: The study results show that exposition to influenza infections 2-4 months prior to birth may be a risk factor in schizophrenia development in adult life. PMID- 15881623 TI - [Psychopharmacotherapy of psychotic disorder during pregnancy and lactation]. AB - The aim of our work based on the global literature-review is to present the guidelines of the psychopharmacotherapy during pregnancy and lactation recommended by various experts-groups as followed: 1. American Academy of Pediatrics, American Psychiatric Association, American College of Obstetricians and Gynecologists, The Maudsley Guidelines, Department of Psychiatry, Medical University of Mainz, Institute of Psychiatry and Neurology in Warsaw. In the first part the guidelines concerning to psychopharmacotherapy of psychotic disorders have been presented and actual data about spreading of these illnesses, pre- and perinatal effects of drugs used, the classification of neuroleptic drugs according to FDA and the safety of these medicines for the breast fed newborn. PMID- 15881624 TI - [An open study on the efficacy and tolerability of quetiapine treatment of patients with schizophrenia]. AB - AIM: Assessment of efficacy and tolerability of quetiapine in treatment of schizophrenia. METHOD: Patients with a diagnosis of schizophrenia (according to ICD-10, DSM-IV) were included in the 12-weeks open study. Severity of schizophrenia symptoms was evaluated using the PANSS Scale. Adverse events have been assessed according to The Adverse Events Inventory, The Simpson Angus Extrapyramidal Symptoms Scale, the Akathisia Barnes Scale and the Involuntary Movement Scale. Quetiapine has been used in increasing doses up to 600 mg/day. RESULTS: Thirty-eight persons were included. The total scores in PANSS before treatment were 84.7 on average. Twenty-eight patients (74%) completed the study. Reduction in severity of schizophrenia according to the PANSS Scale was statistically significant (p < 0.01). Reduction of scores in the range 20-29% was obtained in 18 patients (48.6%), 30-39% in 10 patients (26.3%) and above 40% in 2 patients (5.4%). Adverse symptoms occurred in 71% patients. The most common were: drowsiness (18%), weakness (10.5%), restlessness (10.5%), agitation (10.5%) and increase in severity of delusions and hallucinations (10.5%). CONCLUSION: In the study, Quetiapine caused the reduction in the positive and negative symptoms of schizophrenia. Serious adverse events requiring drop-out of the study were not observed. The severity of extrapyramidal symptoms with statistical significance decreased during the quetiapine treatment (p < 0.05). PMID- 15881625 TI - [Disorder of liver functions in a schizophrenic patient after long-term risperidone treatment--case report]. AB - We present a 51-year old man with an 18-year history of chronic paranoid schizophrenia. The last four years he was treated only with risperidone at a dose of 3 mg per day. He was admitted to hospital due to a psychotic decompensation. Jaundice and skin pruritus were also observed at admission. Laboratory tests revealed elevated liver enzymes. Cholestatic and viral hepatitis were excluded. Risperidone was changed to other antipsychotic drugs. After 7 weeks of treatment the patient was discharged with significant improvement of his mental condition. The liver function test results returned to normal values except for the slightly increased total bilirubin level. Regular liver monitoring was suggested. In this case liver dysfunction was possibly correlated with long-term risperidone therapy. PMID- 15881626 TI - [Clinical example of recovery from psychosis in the context of the theory of psychosis as a state of aberrant salience]. AB - According to the theory proposed by S. Kapur psychosis is a state of aberrant salience. Dopamine and mesolimbic system play the key role in the occurrence of psychosis. According to Kapur dopamine mediates the conversion of neural representations of an external stimulus from a neutral into attractive or aversive. In the psychosis this physiological role of the dopamine is changed. Dopamine starts to create a process of salience acquisition; instead of mediate it as it is in normal circumstances. According to Kapur delusions are cognitive explanations that the individual imposes on the experience of aberrant salience. Pharmacological agents share the psychological effect-dampening salience. But antipsychotics only provide the state of attenuated salience and symptomatic improvement needs further psychological and cognitive resolution. The case of a patient with paranoid psychosis during treatment with risperidone was analyzed in the context of Kapur's theory. Reports of experiences given by patients can support the theory and make it valuable in the conceptualization of the process of step-by-step recovery from psychosis. PMID- 15881627 TI - [Interdependencies between anxiety versus premorbid and present self-image in patients with paranoid schizophrenia]. AB - AIM: The aim of this work was to determine the relationship between anxiety versus premorbid and present self-image in patients with paranoid schizophrenia hospitalized for the first time as well as many a times. TESTED GROUP/MATERIAL: The test group consisted of 120 patients, hospitalized for the first time, aged 19-20 years and diagnosed according to ICD-10 with paranoid schizophrenia, who had suffered from this illness for a period not longer than 2 years. The other group consisted of patients aged 25 - 46, who had suffered from this disease for 5 - 15 years and who had been hospitalized many a times (3 > 10). METHODS: In the tests' the H.G. Gough and A.B. Heilbrunn ACL Adjective test and R. Cattell's Ipat Anxiety Scale were used. The ACL test was done by the patients following the instruction "I am", "before the illness I was". The average, general anxiety level was correlated with the scales of the ACL test of the premorbid image and the present image in both groups. CONCLUSIONS: 1. The anxiety in patients with paranoid schizophrenia is connected both to the premorbid and the present self image. 2. There is a more frequent and stronger relationship between anxiety and present self-images. 3. There are more interdependencies between anxiety and premorbid self-image in patients hospitalized for the first time. 4. There are more interdependencies between anxiety and present self-image in patients with chronic schizophrenia. 5. After the treatment there is no change of the direction of interdependencies but the strength of the relationship is getting deeper, the above tendency being manifested more often and stronger within the chronic group. 6. The higher the anxiety level, the more negative the self-esteem and the greater the need for support, whereas the need for self-accomplishment is lower. PMID- 15881628 TI - [Children from families with mental illness. Case study]. AB - The article reveals the influence of mental illness of one or both parents on the emotional and psychosocial development and possible adjustment disturbances of their children on the example of 3 patients. The patients (all females) were treated in the 4th Psychiatric Clinic of the Institute of Psychiatry and Neurology in Warsaw in the all-day clinic, and in the out-patient clinic with community mobile teams and specialised social help services. The analysis of the history of their mental illnesses was done and compared with the analysis of their family situation and accessible data about the emotional and psychosocial development of their children. Each of these patients was at some time a single parent due to different reasons. In each case the adjustment and emotional disturbances of patients' children occurred on a different level of their development (boy and girl of the primary school age, teenager and young adult). In all presented cases there is a need of help and support apart from the family, which indicates a need of early diagnosis of situations demanding proper systemic interventions. PMID- 15881629 TI - [Social support of chronically mentally ill patients]. AB - AIM: Assessment of the social support system of mentally ill persons covered by a local rehabilitation programme situated in Targowek, Warsaw. METHOD: The sample consists of 92 participants with serious mental illness. Bizon's Social Support Inventory and Social Support Map were used. RESULTS: Patients' social networks were small (9 persons in average) but had a broad scope of functions. Therapists from community rehabilitation services constitute the biggest group of persons included in the individual network of social support. Lack of emotional support is observed. CONCLUSION: Participation in a local system of rehabilitation improves the quantity and quality of individual systems of social support. PMID- 15881630 TI - [Communication and speech disorders and their relationship with psychic development and mental disorders in 8 year old children from the Lodz area]. AB - AIM: The aim of the study was to analyze the occurrence of communication and speech disorders among 8-year-old children in Lodz. The comorbidity of these disorders with mental and somatic disorders was also analysed. We have also studied the correlation between the level of language development and the level of psychic development, and the existence and character of mental disorders. METHOD: The study comprised 7881 children from Lodz, born in 1991. The design of the study was two-stage. Stage I consisted of a screening test, using the Questionnaire of Child-Environment Communication Development, which was sent to the parents of all children. On this basis a group of 58 children with communication disorders was identified, which were further evaluated in the stage II of the study. Stage II consisted of a psychiatric examination, Screening Logopedic Test acc.to Tarkowski, Child Developmental Questionnaire acc. to Rabe Jablonska and Gmitrowicz, somatic state evaluation and analysis of the available documentation. RESULTS: In 0.81% of the children communication disorders were found. Speech disorders were present in all cases: in 2/3rds expressive language disorders or mixed receptive-expressive language disorders, in the remaining cases phonological disorders. Estimated frequency of occurrence of specific disorders in the studied population was as follows: specific developmental language disorders 2.9/1000, acquired aphasia with epilepsy 4/10000, autistic disorder 6.4/10000. Over one third of children with a verbal communication disorder suffered also from various neurological and developmental disorders; most of the children showed abnormal mental development (f = 0.86) and mental disorders (f = 0.66). CONCLUSIONS: Poor language development correlated statistically significantly with mental retardation, pervasive developmental disorder and behaviour disorders, caused by brain damage or brain dysfunction. Children with a communication disorder who demonstrated normal language development suffered from social maladaptation or ADHD statistically significantly more frequently. PMID- 15881631 TI - [Cognitive dysfunction and emotional disturbances in children with neurofibromatosis type one (Recklinghausen's disease). Review of the literature]. AB - Neurofibromatosis type 1 (NF1) is a genetic disorder associated with a high risk of the neuropsychological, emotional and psychosocial disturbances in children with NF1. In this research the results of the psychological and neuropsychological studies in children with NF1 were presented. According to data of current research, the most common cognitive deficits are connected with visual spatial impairment, however verbal, working memory and executive function deficits associated with prefrontal cortex dysfunction are also important. The results indicate that the neuropsychological deficits may increase during the course of the illness. PMID- 15881632 TI - Commentary on Stotz and Griffiths, Burian, and Waters: genes, concepts, DST implications, and the possibility of prototypes. PMID- 15881633 TI - [Medical effects of geomagnetic storms]. PMID- 15881634 TI - [Spiroergometry in elderly patients with CHD]. PMID- 15881635 TI - [Erectile dysfunction syndrome in physician's practice]. PMID- 15881636 TI - [The soluble fibrin monomer in patients with instable angina]. AB - The purpose of the study was to measure the blood level of thrombus precursor protein (TpP), a soluble fibrin monomer, in patients with stable exertional angina (SEA) and healthy people. The study included the examination of 33 patients with SEA (functional class II and III) and 29 practically healthy volunteers (control group). The detection of TpP in blood plasma was performed by solid-phase immune-enzyme analysis ("sandwich" type) using commercial diagnosticum "Kit" ("ABS", USA) and a microplate reader "IEMS Analyzer?Dispenser, with automatic result calculation in "Logistic fif" mode. TpP level in patients with SEA on the average was slightly higher than in control group, but the difference was not significant. TpP blood level was independent of the patients' gender, age, angina functional class and an old myocardial infarction. TpP blood level in patients with SEA depended on the duration of the illness, and proved to be significantly higher (compared with that in control group) in patients with SEA during the first 5 years of the illness, i.e. at early stages of CHD. Solid phase immune-enzyme analysis ("sandwich" type) is a highly informative and affordable clinical method. TpP level in patients with SEA on the average was slightly higher than in healthy people (1.21 +/- 0.06 mkg/ml and 1.01 +/- 0.12 mkg/ml, respectively), but the difference was significant only in patients during the first 5 years of having SEA (1.41 +/- 0.11 mkg/ml). PMID- 15881637 TI - [Cardiac arrhythmias during an elective cholecystectomy]. AB - The paper presents systemized literature data on cardiac arrhythmias (CA) associated with cholecystectomies. The authors present an analysis of CA in 101 patients with chronic calculous cholecystitis (CCC), registered before, during and after an elective cholecystectomy (the surgery was performed either through laparotomy or using endovideosurgical technique). The study revealed that cholecystectomy, especially laparatomic one, bore an arrythmogenic effect. Additional predictors of dangerous CA associated with surgical treatment of CCC, were presence of sinus tachycardia, bradycardia, ciliary arrhythmia, changes in the variability of heart rate, and preoperative heart enlargement. PMID- 15881638 TI - [The influence of nebivolol on thrombocyte aggregation in patients with arterial hypertension with metabolic sydrome]. AB - The research was undertaken with the purpose to assess the efficiency of nebivolol for correction of thrombocyte aggregation capability in patients having arterial hypertension (AH) with metabolic syndrome (MS). 21 patients were administered nebivolol in a dose of 5 mg per day for a month. The dynamic changes in the following parameters were evaluated: anthropometric measurements, blood lipid spectrum, lipid peroxidation in serum and thrombocytes, antioxidative protectability of the liquid part of blood and platelets, thrombocyte aggregation activity. The results were processed using Student criterion and system multifactoral analysis. Nebivolol reduced peroxidation syndrome and optimized thrombocyte aggregation in patients with AH and MS. When administered for a long period of time, nebivolol is able to stabilize the achieved effect. Combining nebivolol application with use of non-drug means is necessary to reduce body weight in patients having AH with MS. PMID- 15881639 TI - [The condition of sympathoadrenal and histaminoreactive systems and the influence of their changes on the peculiarities of a peptic ulcer relapse in patients with myocardial infarction]. AB - The author presents the results of an examination of 106 patients with myocardial infarction (MI) of various severity, who also had a peptic ulcer relapse. Patients with moderate and severe MI in the majority of cases had solitary duodenal ulcers whereas those with extremely severe MI often had multiple ulcers in the body and the cardial part of the stomach, which were bleeding in 43.5% of cases. The pathogenetic basis of the ulcerogenesis was focal microcirculation disorders in the mucosa of the gastroduodenal zone of thrombohemorragic or thromboischemic type. In patients with moderate or severe MI they appeared against the background of hemostatic disturbances typical of the first stage of thrombohemorragic syndrome (hypercoagulation). In extremely severe MI they corresponded to the second stage i.e. partial consumption coagulopathy. In patients with moderate coronary pathology these changes were promoted by the activation of acid-peptic factor, the depression of mucopolysaccharide production, hypo- and hyperkinetic dyskinesis, and in those with extremely severe coronary pathology --the depression of all gastric function except normal acid production. One of the trigger mechanisms of ulcerogenesis could be changes in the activity of sympathoadrenal and histaminereactive systems, which increased in cases of moderate and severe MI and decreases to a certain extent in cases of extremely severe MI. PMID- 15881640 TI - [The functional condition of the cardiovascular system and adaptation abilities in patients with calculous cholecystitis]. AB - Clinical and functional evaluation of the cardiovascular system in patients with various clinical course of chronic calculous cholecystitis was carried out in pre and postoperative period under conditions of muscle rest and allowable physical activity using treadmill test. Physical activity modeling found a range of potential complications, which are most characteristic of elderly patients as well as patients with coronary heart disease and postinfarction cardiosclerosis (PC). The study shows that operative intervention in patients with PC should be scheduled not only with the remoteness of the old myocardial infarction taken into account, but also considering the degree to which the heart has preserved its capability of adapting to stressful situations. PMID- 15881641 TI - [Myocarditis in severe typhoid fever]. AB - The article explains the development of toxic infective myocarditis and, in many cases, pancarditis, in patients with severe typhoid fever (TF) with lethal outcome. The authors present the most frequently found symptoms and ECG signs, the main histological features and possibilities of clinical diagnostics of myocarditis. The latter is of special importance, because a significant number of patients do not have typical clinical presentation. The work is based on the data from 109 autopsy records and 42 case histories as well as the results of morphological study of the heart in archive samples received from 47 military men who died of TF. PMID- 15881642 TI - [Possibilities of Doppler echocardiography application in a patient on program hemodialysis]. AB - The aim of the study was to investigate the significance of Doppler echocardiography (echoCG) for assessment of "dry weight" in hemodialysis patients. 43 dialysis patients (20 men and 23 women aged 49 +/- 11 years), receiving 4-hour bicarbonate hemodialysis (HD) 3 times a week, were studied prior to and after HD procedures. M-mode echoCG was performed and left ventricular mass index (LVMI) and ejection fraction (EF) were calculated. Transmitral flow was assessed by Doppler echoCG. Peak velocity of early (E) and late (A) filling, E/A ratio, isovolumic relaxation time (IVRT) and early deceleration time (DT) were detected. Left ventricular hypertrophy (LVH) was detected in 37 (86.0%) patients. EF was lower than 45% in 4 patients. There was significant positive correlation between the amount of ultrafiltration (evaluated in per cents of body mass after HD) and deltaE (r = 0.59; p = 0.001), and there was found no correlation between the amount of ultrafiltration and deltaA. Patients with intradialytic hypotension had substantially higher DT than patients without one (238.7 +/- 64.3 vs. 168.6 +/- 51.2 mc, respectively, p < 0.001). "Dry weight" is associated with increase of early DT over the level normal for the age in hemodialysis patients with left ventricular hypertrophy. Patients with increased early DT are exposed to the high risk of intradialytic hypotension development. PMID- 15881643 TI - [Diagnostics and therapeutic approach in clinical remission of chronic bronchitis]. AB - Clinical examination and laboratory tests performed on 708 patients with chronic obstructive and non-obstructive bronchitis (COB and CNB), demonstrated heterogeneous character of the group of these patients in clinical remission. This suggests that it is appropriate to distinguish between the stages of complete and partial remission, implying different tasks concerning treatment and rehabilitation. Remission of CNB is characterized by minimal deviation of clinical and laboratory parameters; functional balance between the lipid peroxidation-antioxidative activity (LP-AA) system and immunity is preserved. At the stage of rehabilitation of these patients the major attention is paid to the general sanitary measures (physiotherapy exercises, climate therapy). The stage of partial remission in COB and CNB is characterized by residual activity of the inflammatory process, a disbalance between the LP-AA system and immunity, most prominent in partial remission of COB. Rehabilitation of these patients, in addition to physiotherapy exercises and tempering of the organism, should include special activity, directed towards the normalization of LP and immunity parameters. PMID- 15881644 TI - [Effectiveness of metabolic preparations in complex treatment of elderly patients with postinfarction cardiosclerosis and circulatory insufficiency]. AB - The paper presents the results of an examination of 129 elderly patients with CHD who had had a myocardial infarction and suffered from II-III functional class heart failure. The patients were divided into four groups. The patients in the first group were administered complex therapy (CT) only; the second group received CT plus a composition of replaceable amino acids (CRAA) sublingually and orally, the third group received CT plus a placebo and the fourth--CT plus melatonin in doses of 3 and 6 mg. The efficiency of the treatment was assessed by the dynamics of clinical manifestations and by means of echoCG and Holter ECG monitoring; parameters of lipid peroxidation and antioxidative protectability in erythrocytes were measured. The comparison of the results suggests that addition of CRAA and melatonin in a dose of 6 mg to CT is more effective than CT only. This was proved by the more pronounced positive dynamics of clinical symptoms, the improval of the myocardial contractility and normolizing influence on the balance in the oxidant/antioxidant system. PMID- 15881645 TI - [Helicobacter pylori infection in patients with chronic hives and asthma]. AB - Difficulties in treatment of allergic diseases (ADs) are often determined by the fact that the origin of the illness is impossible to establish. Foreign researchers have demonstrated a correlation between Helicobacter pylori (HP) persistence and such conditions as chronic relapsing urticaria (CRLU), Quincke's edema (QO) and respiratory manifestations of allergy (usually bronchial asthma (BA)). HP may participate in the forming of ADs in three possible ways: 1) the bacteria interact with mast cells, and thus are able to initiate mediator liberation; 2) being full antigens, the bacteria themselves are able to cause allergy; 3) the infection process development impairs the barrier function of the alimentary tract mucosa, thus impairing food processing. This creates conditions for allergic food particles to enter bloodstream, which is facilitated by inflammatory lesions of the intestinal tract, protozoal and helminthic invasion, and dysbacteriosis. The aim of the study was to determine how frequently patients with CRU, QO and BA have HP invasion and develop intestinal microflora disbalance, as well as to find out whether there is a correlation between HP infection and alterations in the IgE-system, and establish anti-HP therapy effects, measuring the levels of anti-helicobacter antibody and IgE before and after the treatment. The study revealed HP invasion in 89.2% of patients with chronic ADs; 96.7% of the subjects had disturbances of microbiocenosis; total IgE level correlated with the allergic process activity and anti-helicobacter antibody level; eradicational therapy was more effective when included proton pump inhibitor, clarithromycin and flemoxin than when de-nol, clarithromycin and flemoxin were administered. According to the results of the study, anti helicobacter therapy allows more effective treatment of patients with relapsing ADs and HP infection. PMID- 15881646 TI - [Features of gastroesophageal reflux disease in young people]. AB - The article contains ethiopathogenic, clinical and diagnostic criteria of gastroesophageal reflux disease (GERD) in young people. GERD verification was based upon the results of the examination of 137 men aged 18 to 23 years. These results included clinical findings as well as the data from the following tests: intraesophageal and intragastric pH-metry with evaluation of parietal cell response to atropine, and endoscopic and morphological examination of the upper division of the gastrointestinal tract including detection of Helicobacter pylori (HP) expansion, evaluation of inflammatory and regenerative processes in the mucosa of the lower third of the esophagus and the antral part of the stomach. The results show that the feature of GERD in the given age group is a certain morphological picture in the esophagus characterized by a various degree of squamous epithelium dystrophy with almost no metaplasia, as well as a high degree of HP adhesion in the esophagus with atropine-resistent hyperchlorhydria in the stomach. It shoud be noted that eradicational and antisecretory therapy in young people with GERD leads to the reestablishment of the esophagial mucosa structure. PMID- 15881647 TI - [Complex approach in excessive body mass and obesity correction]. PMID- 15881648 TI - [Duodenal leiomyoma]. PMID- 15881649 TI - Radiation-induced bystander effect and adaptive response in mammalian cells. AB - Two conflicting phenomena, bystander effect and adaptive response, are important in determining the biological responses at low doses of radiation and have the potential to impact the shape of the dose-response relationship. Using the Columbia University charged-particle microbeam and the highly sensitive AL cell mutagenic assay, we show here that non-irradiated cells acquire mutagenesis through direct contact with cells whose nuclei have been traversed with a single alpha particle each. Pretreatment of cells with a low dose of X-rays four hours before alpha particle irradiation significantly decreased this bystander mutagenic response. Results from the present study address some of the fundamental issues regarding both the actual target and radiation dose effect and can contribute to our current understanding in radiation risk assessment. PMID- 15881650 TI - Anticarcinogenic effect of Nymphaea alba against oxidative damage, hyperproliferative response and renal carcinogenesis in Wistar rats. AB - The present study investigates the prophylactic effect of Nymphaea alba against ferric nitrilotriacetate (Fe-NTA)-induced renal oxidative stress, hyperproliferative response and renal carcinogenesis in Wistar rats. Treatment with Fe-NTA (9 mg Fe/kg body weight, intraperitoneally) enhanced iron-ascorbate induced renal lipid peroxidation, xanthine oxidase, gamma-glutamyl transpeptidase and hydrogen peroxide (H2O2) generation with reduction in renal glutathione content, antioxidant enzymes, viz., glutathione peroxidase, glutathione reductase, catalase, glucose-6-phosphate dehydrogenase and phase-II metabolising enzymes such as glutathione-S-transferase and quinone reductase. It also elevated the levels of blood urea nitrogen, serum creatinine, ornithine decarboxylase (ODC) activity and thymidine [3H] incorporation into renal DNA. It also enhanced DEN-initiated renal carcinogenesis by increasing the percentage incidence of renal tumors. Treatment of rats orally with N. alba (100 and 200 mg/kg body weight) resulted in significant decrease in gamma-glutamyl transpeptidase, lipid peroxidation, xanthine oxidase, H2O2 generation, blood urea nitrogen, serum creatinine, renal ODC activity, DNA synthesis (p < 0.001) and incidence of tumors. Renal glutathione content (p < 0.01), glutathione metabolizing enzymes (p < 0.001) and antioxidant enzymes were also recovered to significant level (p < 0.001). Thus, our results show that N. alba is a potent chemopreventive agent and suppresses Fe-NTA-induced oxidative stress, hyperproliferative response and renal carcinogenesis in Wistar rats. PMID- 15881651 TI - Protein synthesis and urokinase mRNA metabolism. AB - Expression of the urokinase-type plasminogen activator (uPA) is under tight regulation by hormones, cytokines and growth factors under physiological conditions. Treatment of lung epithelial (Beas2B) cells with translation inhibitors induces uPA mRNA expression, as well as early response genes. To understand the specific expression and regulation of uPA mRNA, we treated Beas2B cells with cycloheximide (CycD), anisomycin, emitine and puromycin in a time dependent manner and measured uPA mRNA expression by Northern blotting. All these agents induced uPA mRNA by two- to seven-fold within 3 h after treatment in Beas2B cells. CycD, emitine, puromycin and anisomycin also enhanced uPA mRNA half life by three- to five-fold in Beas2B cells treated with DRB, an inhibitor of transcription. However, run-on-transcription experiments indicated that these agents failed to induce uPA mRNA transcription indicating that they augment uPA mRNA mainly due to increased stability. Using gel mobility shift, we identified an uPA mRNA binding protein (uPA mRNABp) that selectively binds to uPA mRNA [Gyetko MR, Todd III RF, Wilkinson CC, Sitrin RG: The urokinase receptor is required for human monocyte chemotaxis in vitro. J Clin Invest 93: 1380-1387, 1994]. Binding of both cytoplasmic and nuclear uPA mRNABp to uPA mRNA was abolished after treatment with translation inhibitors, which coincides with the maximal expression of uPA mRNA. We also found a similar decline in HuR and heterogeneous nuclear ribonucleoprotein C (hnRNPC) which are known to stabilize uPA mRNA both in the nuclear and cytosolic compartments. These results strongly suggest that increased uPA mRNA stability induced by translational inhibitors involves the interaction of uPA mRNA with a degrading protein factor rather than increased interaction of proteins that are known to stabilize uPA mRNA. These data also strongly suggests that down-regulation of the uPA-uPA mRNABp interaction by translational inhibitors rather than the translocation of uPA mRNABp contributes to increased uPA mRNA stability. This pathway may regulate uPA mediated functions of the lung epithelium in the context of inflammation or neoplasia. PMID- 15881652 TI - Tsc-22 enhances TGF-beta signaling by associating with Smad4 and induces erythroid cell differentiation. AB - Tsc-22 was isolated as a TGF-beta-inducible gene by differential screening of the mouse osteoblastic cell cDNA library [J Biol Chem 267 (1992) 10219]. tsc-22 mRNA is expressed in almost all organs of mice and humans and its expression is induced in a variety of cell lines by many different factors including TGF-beta, phorbol ester, serum, and progestin. tsc-22 encodes a 18-kd protein that contains a leucine zipper motif and a Tsc-box. The leucine zipper motif of the Tsc-22 protein does not have a basic DNA binding motif and when the protein was fused to a heterologous DNA binding domain, it showed various transcription-modulating activities ranging from activation to repression [J Biol Chem 274 (1999) 27439, Biochem Biophys Res Commun 278 (2000) 659]. Although these results suggest that the Tsc-22 protein functions as a transcriptional regulator recruiting various coactivators or repressors, its mechanism is not known. In this study, we examined whether Tsc-22 modulates the TGF-beta-dependant signaling pathway and found that Tsc-22 binds to and modulate the transcriptional activity of Smad3 and Smad4. Its effect on cellular differentiation was also examined. PMID- 15881653 TI - Threshold levels of ERK activation for chemotactic migration differ for NGF and EGF in rat pheochromocytoma PC12 cells. AB - In a previous study, we show that stimulation of chemotaxis in rat pheochromocytoma PC12 cells by nerve growth factor (NGF) and epidermal growth factor (EGF) requires activation of the RAS-ERK signaling pathway. In this study, we compared the threshold levels of ERK activation required for EGF and NGF stimulated chemotaxis in PC12 cells. The threshold ERK activity required for NGF to stimulate chemotaxis was approximately 30% lower than that for EGF. PD98059 treatment inhibited EGF stimulation of growth and chemotaxis; however, stimulation of chemotaxis required an EGF concentration approximately 10 times higher than for stimulation of PC12 cell growth. Thus, ERK-dependent cellular functions can be differentially elicited by the concentration of EGF. Also, treatment of PC12 cells with the PI3-K inhibitor LY294002 reduced ERK activation by NGF; thus, higher NGF concentrations were required to initiate chemotaxis and to achieve the same maximal chemotactic response seen in untreated PC12 cells. Therefore, the threshold NGF concentration to stimulate chemotaxis could be adjusted by the crosstalk between the ERK and PI3-K pathways, and the contributions of PI3-K and ERK to signal chemotaxis varied with the concentrations of NGF used. In comparison, LY294002 treatment had no effect on ERK activation by EGF, but the chemotactic response was reduced at all the concentrations of EGF tested indicating that NGF and EGF differed in the utilization of ERK and PI3-K to signal chemotaxis in PC12 cells. PMID- 15881654 TI - Ascorbate uptake in pig coronary artery endothelial cells. AB - Although smooth muscle and endothelial cells in pig coronary artery are morphologically and functionally distinct, ascorbate uptake has been characterized only in smooth muscle cells. Ascorbate transporters in kidney and intestinal epithelial cells differ from those in smooth muscle. We examined ascorbate transport and mRNA expression of sodium-dependent vitamin C transporters (SVCT) by RT-PCR in the pig coronary artery endothelial cell cultures. When 14C-ascorbate uptake in endothelial cells was examined as 14C or by HPLC, the two values did not differ from each other. 14C-ascorbate uptake was Na(+)-dependent, stereoselective for L-ascorbate and inhibited by sulfinpyrazone. The kinetic characteristics of the uptake were: Km = 27 +/- 3 microM (Hill coefficient = 1) for ascorbate and Km = 73 +/- 14 mM (Hill coefficient = 2) for Na+. Surprisingly, endothelial cells had similar kinetic parameters as smooth muscle cells, except for a slightly lower uptake velocity in endothelial cells. Comparison with the smooth muscle showed that both tissue types expressed mRNA for SVCT2. Endothelial cells differ from epithelial cells which express mainly SVCT1 but resemble smooth muscle cells in this respect. PMID- 15881655 TI - External bioenergy-induced increases in intracellular free calcium concentrations are mediated by Na+/Ca2+ exchanger and L-type calcium channel. AB - External bioenergy (EBE, energy emitted from a human body) has been shown to increase intracellular calcium concentration ([Ca2+]i, an important factor in signal transduction) and regulate the cellular response to heat stress in cultured human lymphoid Jurkat T cells. In this study, we wanted to elucidate the underlying mechanisms. A bioenergy specialist emitted bioenergy sequentially toward tubes of cultured Jurkat T cells for one 15-minute period in buffers containing different ion compositions or different concentrations of inhibitors. [Ca2+], was measured spectrofluorometrically using the fluorescent probe fura-2. The resting [Ca2+]i in Jurkat T cells was 70 +/- 3 nM (n = 130) in the normal buffer. Removal of external calcium decreased the resting [Ca2+]i to 52 +/- 2 nM (n = 23), indicating that Ca2+ entry from the external source is important for maintaining the basal level of [Ca2+]i. Treatment of Jurkat T cells with EBE for 15 min increased [Ca2+]i by 30 +/- 5% (P < 0.05, Student t-test). The distance between the bioenergy specialist and Jurkat T cells and repetitive treatments of EBE did not attenuate [Ca2+]i responsiveness to EBE. Removal of external Ca2+ or Na+, but not Mg2+, inhibited the EBE-induced increase in [Ca2+]i. Dichlorobenzamil, an inhibitor of Na+/Ca2+ exchangers, also inhibited the EBE induced increase in [Ca2+]i in a concentration-dependent manner with an IC50 of 0.11 +/- 0.02 nM. When external [K+] was increased from 4.5 mM to 25 mM, EBE decreased [Ca2+]i. The EBE-induced increase was also blocked by verapamil, an L type voltage-gated Ca2+ channel blocker. These results suggest that the EBE induced [Ca2+]i increase may serve as an objective means for assessing and validating bioenergy effects and those specialists claiming bioenergy capability. The increase in [Ca2+]i is mediated by activation of Na+/Ca2+ exchangers and opening of L-type voltage-gated Ca2+ channels. PMID- 15881656 TI - Plasma bikunin: half-life and tissue uptake. AB - Bikunin is a chondroitin sulfate-containing plasma protein synthesized in the liver. In vitro, it has been shown to inhibit proteases and to have additional activities, but its biological function is still unclear. Here we have studied the dynamics of plasma bikunin in rats and mice. A half-life of 7 +/- 2 min was obtained from the time course of the decrease of the plasma level of bikunin following hepatectomy. Clearance experiments with intravenously injected radiolabeled bikunin with or without the chondroitin sulfate chain showed that the polysaccharide had little influence on the elimination rate of the protein. The uptake of bikunin by different tissues was studied using bikunin labeled with the residualizing agent 125I-tyramine cellobiose; 60 min after intravenous injection, 49% of the radioactivity was recovered in the kidneys and 6-11% in the liver, bones, skin, intestine and skeletal muscle. The uptake in the liver was analyzed by intravenous injection of radiolabeled bikunin followed by collagenase perfusion and dispersion of the liver cells. These experiments indicated that bikunin is first trapped extracellularly within the liver before being internalized by the cells. PMID- 15881657 TI - Altered lipid profile and changes in uroplakin properties of rat urothelial plasma membrane with diets of different lipid composition. AB - Rigid plaques containing protein particles in plasma membrane build on the apical surface of the mammalian urothelium. We have previously shown that dietary fats modified the fatty acid profile as well as the fluorescence anisotropy of rat urothelial plasma membranes. In this study, we have further examined the proportion of phosphatidylcholine, phosphatidylethanolamine, cerebrosides, sulfatides and cholesterol in detergent resistant (DRM) and soluble (DSM) plasma membrane fractions as well as the properties of the particles. Four groups of weaned rats were fed for 12 weeks on a commercial diet (control), or on a formula containing 5% (w/w) of corn oil, fish oil or olein. The control DRM behaved as a distinctive domain since it was enriched in cholesterol and glycosphingolipids. DSM showed higher levels of phosphatidylcholine and phosphatidylethanolamine with respect to DRM. On the other hand, the lipid distributions were affected by the diets. Homogeneous lipid distributions between DSM and DRM were found in olein membranes, suggesting a decreased potential formation of lipid domains. In addition, properties of the uroplakins were altered by dietary treatments. Thus, uroplakins (UP) Ia, Ib, II and III observed by SDS-PAGE, were in lower proportions (mainly olein) than in controls. Moreover, a higher proportion of UPIII was cross-linked to UPIII and UPlb in olein treatment than in control. Meanwhile, only cross-linking to UPIII or UPIb was altered in corn and fish diets, respectively. These results suggest a role of the lipids in the establishment of the uroplakin interactions. Thus, specific dietary fats may have important functional implications. PMID- 15881658 TI - Angiotensin II stimulated transcription of cyclooxygenase II is regulated by a novel kinase cascade involving Pyk2, MEKK4 and annexin II. AB - Although it is known that MEKK4 regulates MKK6, and p38 MAP kinase, extracellular stimuli that activate the serine/threonine kinase, MEKK4, are unknown. The aim of this study was then to identify stimuli that regulate MEKK4. By using recombinant MEKK4, as bait to attract interacting proteins, the calcium binding protein, annexin II, was identified by mass spectrometry as interacting with MEKK4, suggesting that MEKK4 might be regulated by calcium. A calcium-dependent interaction between MEKK4 and annexin II was observed when MEKK4 was immunoprecipitated from rat aortic smooth muscle cells that were treated with angiotensin II. Additional studies using recombinant MEKK4 in a Far-Western immunoblot identified a protein of 120 kDa as interacting directly with MEKK4. Prior studies indicated that MEKK4 was phosphorylated on tyrosine in vivo, and in fact, Pyk2 interacts with MEKK4 in an angiotensin II dependent manner in rat aortic smooth muscle cells. Pyk2 phosphorylates MEKK4 in vitro and Pyk2-dependent phosphorylation further regulates MEKK4-dependent phosphorylation of MKK6. Finally, dominant-negative MEKK4 inhibits angiotensin II mediated transcription of a luciferase reporter construct containing the cyclooxygenase II promoter, demonstrating that MEKK4 functions in a calcium-dependent manner as a substrate for Pyk2 and regulates transcription of cyclooxygenase II. PMID- 15881659 TI - Expression pattern of voltage-dependent calcium channel alpha1 and beta subunits in adrenal gland of N-type Ca2+ channel alpha1B subunit gene-deficient mice. AB - The Ca2+ channel alpha1B subunit is a pore-forming component capable of generating N-type Ca2+ channel activity. Although N-type Ca2+ channel plays a role in a variety of neuronal functions, alpha1B-deficient mice exhibit normal life span without apparent abnormalities of behavior, histology or plasma norepinephrine level, presumably owing to compensation by some other Ca2+ channel alpha1 or beta subunit. In this study, we studied the levels of alpha1A, alpha1C, alpha1D, C1E, beta1, beta2, beta3 and beta4 mRNAs in adrenal gland of alpha1B deficient mice. The alpha1A mRNA in homozygous mice was expressed at higher level than in wild or heterozygous mice, but no difference in the expression levels of alpha1c, alpha1D, alpha1E, beta1, beta2, beta3 and beta4 was found among wild, heterozygous and homozygous mice. The protein level of alpha1A in homozygous mice was also expressed at higher level than in wild or heterozygous mice. To examine whether increased expression is induced by cis-regulatory element within 5' upstream region of alpha1A gene, we examined lacZ expression in alpha1B-deficient x alpha1A6.3-lacZ mice (carrying a 6.3-kb 5'-upstream fragment of alpha1A gene fused to E. coli lacZ reporter gene), which express lacZ in medullar chromaffin cells, but not in cortex. The levels of lacZ expression in homozygous alpha1B deficient x alpha1A6.3-lacZ mice were higher than in wild or heterozygous mice. Therefore, a possible explanation of the normal behavior and plasma norepinephrine level of alpha1B-deficient mice is that compensation by alpha1A subunit occurs and that 6.3-kb 5'-upstream region of alpha1A gene contains enhancer cis-element(s) for compensation in adrenal medulla chromaffin cells. PMID- 15881660 TI - Chemopreventive effect of piperine on mitochondrial TCA cycle and phase-I and glutathione-metabolizing enzymes in benzo(a)pyrene induced lung carcinogenesis in Swiss albino mice. AB - Piperine is a major component of black (Piper nigrum Linn) and long pepper (Piper longum Linn) used widely in various systems of traditional medicine. We have evaluated the effect of piperine on mitochondrial tricarboxylic acid cycle and phase I and glutathione-metabolizing enzymes in Benzo(a)pyrene induced experimental lung carcinogenesis in swiss albino mice. Lung cancer bearing mice showed a significant decrease in the activities of mitochondrial enzymes isocitrate dehydrogenase (ICDH), -ketoglutarate dehydrogenase (KDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH) and significantly increased NADPH Cytochorome reductase (NADPH-C reductase), cytochrome P450 (cyt-p450) and cytochrome b5(cyt-b5). The activities of glutathione-metabolizing enzymes glutathione peroxidase(GPx), glutathione reductase (GR) and glucose-6-phospho dehydrogenase(G6PDH) were significantly lowered in lung-cancer bearing mice when compared with control mice. Piperine supplementation to tumour-induced animals significantly lowered the phase-I enzymes (NADPH-C reductase, cyt-p450 and cyt b5)) and there was a rise in glutathione-metabolizing enzymes (GPx, GR and G6PDH), which indicated an antitumour and anti-cancer effect. Comparison of normal control mice and mice administered piperine only as drug control showed no significant variations in enzyme activities. Piprine administration to benzo(a)pyrene induced animals significantly increased the activities of mitochondrial enzymes, thereby suggesting its role in mitochondrial energy production. PMID- 15881661 TI - The effects of nitric oxide synthesis on the Na+ ,K(+)-ATPase activity in guinea pig kidney exposed to lipopolysaccharides. AB - Endotoxins (lipopolysaccharides; LPS) are known to cause multiple organ failure, including renal dysfunction. LPS triggers the synthesis and release of cytokines and the vasodilator nitric oxide (NO*). A major contributor to the increase in NO* production is LPS-stimulated expression of inducible nitric oxide synthase (iNOS). This occurs in vasculature and most organs including the kidney. During endotoxemia, NO* and superoxide react spontaneously to form the potent and versatile oxidant peroxynitrite (ONOO-) and the formation of 3-nitrotyrosine (nTyr)-protein adducts is a reliable biomarker of ONOO- generation. Therefore, the present study was aimed at investigating the role of endogenous nitric oxide in regulating Na+,K(+)-ATPase activity in the kidney, and at investigating the possible contribution of reactive nitrogen species (RNS) by measuring of iNOS activity. In addition, the present study was aimed at investigating the relationship between nTyr formation with iNOS and Na+,K(+)-ATPase activities. Previously in our study, nTyr was not detectable in kidney of normal control animals but was detected markedly in LPS exposed animals. In this study, kidney Na+,K(+)-ATPase activity were maximally inhibited 6 h after LPS injection (P:0.000) and LPS treatment significantly increased iNOS activity of kidney (P:0.000). The regression analysis revealed a very close correlation between Na+,K(+)-ATPase activity and nTyr levels of LPS treated animals (r = -0.868, P = 0.001). Na+,K(+)-ATPase activity were also negatively correlated with iNOS activity (r = -0.877, P = 0.001) in inflamed kidney. These data suggest that NO* and ONOO- contribute to the development of oxidant injury. Furthermore, the source of NO* may be iNOS. iNOS are expressed by the kidney, and their activity may increase following LPS administration. In addition, NO* and ONOO- formation inhibited Na+,K(+)-ATPase activity. This results also have strongly suggested that bacterial LPS disturbs activity of membrane Na+,K(+)-ATPase that may be an important component leading to the pathological consequences such as renal dysfunction in which the production of RNS are increased as in the case of LPS challenge. PMID- 15881662 TI - Altered structure and expression of RB1 gene and increased phosphorylation of pRb in human vestibular schwannomas. AB - Tumor-specific alterations at the RB1 gene locus in 30 human vestibular schwannomas including 10 NF2 and 20 sporadic cases were analysed. Southern blot analysis of DNA from these samples revealed loss of heterozygosity (LOH) at the RB1 locus in 6 of 24 informative cases (25%) compared to normal blood DNAs from the same patients. Northern blot analysis showed normal size RB1 mRNA in all the tumor samples. However, there was a 2-5-fold increase in the level of expression of the RB1 gene in all the tumor samples compared to the WI38 cell line which was used as control. Western blot analysis of the RB1 protein, pRb showed a 2.5-5 fold increase in the level of total pRb as compared to normal WI38 cell line. Sixty five to seventy five percent of the total pRb were in phosphorylated form in most tumors. The LOH at the RB1 gene locus suggests genetic instability in these patients. Further, increased levels of RB1 mRNA, total pRb and the phosphorylated form of pRb suggests that RB1 gene in these tumors may have anti apoptotic function. These results suggest that the RB1 gene has a major role in the development of human vestibular schwannomas. PMID- 15881663 TI - Control of scavenger receptor-mediated endocytosis by novel ligands of different length. AB - The scavenger receptor recognized as a multiligand family of receptors falls in the group that is internalised through endocytosis. In this report we used several recombinant fragments of the tapeworm protein paramyosin, known to form filamentous dimers that bind collagenous structures as ligands of different length for the class A type I scavenger receptor (SR-AI). While native CHO cells are unresponsive to any of the recombinant fragments, it is shown that CHO cells transfected with this receptor efficiently internalise recombinant fragments that correspond to two thirds of the full-length paramyosin. In contrast, recombinant products corresponding to one-third of the full-length paramyiosin are not internalised. It is also shown that important molecules in the organization of the coated pit, are enriched when the two-thirds long paramyosin fragments were bound and internalised through the SR-AI. Moreover, internalisation of these fragments trigger a classical apoptotic pathway shown by the presence of TUNEL positive cells and the appearance of apoptotic bodies. We report paramyosin as a new ligand for the scavenger receptor and provide evidence supporting the notion that these receptors upon the formation of arrays with length-specific molecules, not only trigger endocytosis but also seem to regulate the synthesis of molecules involved in the organization of coated pits. PMID- 15881664 TI - Effect of ethanol on the response of the rat urinary bladder to in vitro ischemia: protective effect of alpha-lipoic acid. AB - PURPOSE: Ethanol exposure has been used to demonstrate the increase of oxidative stress to a variety of tissues. We studied the effect of ethanol on the response of isolated strips of rat bladder to in vitro hypoxia in the absence of glucose (in vitro ischemia). Secondly, we determined if alpha-lipoic acid (LA) could alter the response to ethanol + in vitro ischemia. METHODS: Sixty-four rats were used for the these experiments. Each rat was anesthetized and its urinary bladder excised. The bladder body was cut into two longitudinal strips and each strip mounted in individual baths filled with oxygenated Tyrodes solution containing glucose at 37 degrees C. Ethanol (0.3%, 1%, or 3%) was placed in the first six baths (two strips at each concentration). The last two baths did not receive ethanol. Each strip was incubated for 1 h and then stimulated with field stimulation at 2, 8, and 32 Hz. Each strip was stimulated with 10 microM carbachol, washed three times with fresh oxygenated buffer and ethanol re-added to their respective baths. Each strip was then stimulated with 120 mM KCl and washed three times as before. Strips were then subjected to 1 h in vitro ischemia (incubation in the absence of glucose with Tyrode's equilibrated with nitrogen instead of oxygen). During the ischemic period, each strip was stimulated for 5 s every 10 min by 32 Hz FS to simulate hyperreflexia. At the end of the hour, the tissues were incubated for an additional hour in the presence of oxygen + glucose and subjected to a second series of stimulations as before. At all times, ethanol was maintained in baths 1-6. In set 2, 1% ethanol was added to the first six baths. LA was added to every other bath, and the experiments performed as mentioned earlier. RESULTS: (a) Ethanol at 0.3% or 1% had no effect on the contractile responses prior to exposure to in vitro ischemia; 3% was inhibitory. (b) In vitro ischemia mediated a significant decrease in the contractile responses to all forms of stimulation except for carbachol. (c) Ethanol mediated a dose-response enhancement of the contractile dysfunctions caused by in vitro ischemia. (d) LA completely reversed the effects of ethanol on contractile responses following in vitro ischemia except for carbachol. CONCLUSIONS: The results demonstrate that direct exposure to ethanol significantly enhanced contractile dysfunctions mediated by in vitro ischemia followed by re-oxygenation and that the presence of LA significantly inhibits this effect of ethanol. PMID- 15881665 TI - Radiomodulatory effect of liposome encapsulated AK-2123 on tumor in mice exposed to hepatocarcinogen. AB - An attempt was made to evaluate the whole body gamma-radiation effect on tumor in the presence of free and liposome encapsulated AK-2123, a hypoxic cell radiosensitizer that has widely been used in combination with a number of cancer therapies such as thermotherapy, chemotherapy and radiotherapy. Entrapment efficiency of AK-2123 into liposome was determined by LASER Raman spectroscopy. Cancer induction in mice was carried out by repeated exposure of N nitrosodiethylamine (DEN) in combination with partial hepatectomy. Parameters such as marker enzymes activities (GGT and AChE), rates of nucleic acid synthesis, viability modification factor and the histology of liver tissues monitored, supported the induction of cancer in liver. In addition, the effect of free as well as liposome encapsulated AK-2123 on haemopoietic parameters were also studied. It was observed that AK-2123 after incorporation into liposome afforded more efficient radiomodulatory effects than that of free AK-2123 as determined by the above-mentioned parameters. Neither free AK-2123 nor liposome encapsulated AK-2123 showed any detectable toxic effects on the mice. Thus, it is seen that treatment of cancer with a combination of radiation, a radiomodifier and a drug delivery system, opens a wide scope for exploitation for the improvement of existing cancer therapies. PMID- 15881666 TI - P5644 interacts with phosphatidylinositol-4-phosphate adaptor protein-1 associated protein-1. AB - The human novel gene pp5644 (GeneBank Accession No. AF289559) coding for 124 amino acids was recently cloned. Overexpression of pp5644 in Hela cells significantly inhibited the growth and colony formation. The pp5644-interacting protein FAPP1 (phosphatidylinositol-four-phosphate adaptor protein1) associated protein-1, called FASP1, was obtained by using yeast two-hybrid system. The interaction between pp5644 and FASP1 was experimentally confirmed by GST pull down assay in vitro and co-immunoprecipitation assay in vivo. Co-localization of pp5644 and FASP1 in cytoplasm in Hela cells could further support the interaction. Based on the experimental results, it is suggested that pp5644 physically bind to FASP1 and the biological significance of this kind of interaction in vivo is discussed. PMID- 15881667 TI - Effect of mercuric chloride on various hydroxyproline fractions in rat serum. AB - Mercuric chloride (HgCl2) disturbs the collagen metabolism in the body which is reflected by altered hydroxyproline fractions in the serum. The aim of the present investigation was to study the effect of HgCl2 treatment on various hydroxyproline (Hyp) fractions in rat serum and the effect of 2,3-dimercapto-1 propane sulfonic acid (DMPS) treatment on serum Hyp fractions in HgCl2 treated rats. Other parameters studied included body weight, food intake, water intake and kidney weight. Doses of HgCl2 used were 0.1, 0.5, 1.0, 2.0, 3.0 mg/kg body weight and that of DMPS was 100 mg DMPS/kg body weight. All the doses of HgCl2 used caused significant (p < 0.01) alterations in free, peptide-bound and protein bound Hyp in the serum when compared with control rats but a dose of 2 mg/kg body weight caused significant (p < 0.001) alteration even in the total serum Hyp when compared to control rats. Administration of DMPS prior HgCl2 treatment of rats sacrificed 24 h after the treatment caused a significant decrease of 52% (p < 0.01) in free Hyp when compared to similar HgCl2 treated rats. DMPS treatment with HgCl2 also caused an increase of 61% (p < 0.001) and 114% (p < 0.001) in peptide- and protein-bound Hyp respectively, when compared to HgCl2 treated rats sacrificed 24 h after mercuric chloride and DMPS treatment. Administration of DMPS followed by HgCl2 to rats which were sacrificed 48 h later caused no significant change in the total and free Hyp when compared to HgCl2 treated rats which were sacrificed 48 h after the treatment. But there was a significant decrease of 40% (p < 0.001) in peptide-bound Hyp and an increase in of 77% (p < 0.001) in protein-bound Hyp when compared to HgCl2 treated rats sacrificed 48 h after the treatment. The present study shows that HgCl2 treatment caused significant alterations in serum Hyp fractions reflecting disturbed composition of connective tissues which were not reversed by DMPS treatment. PMID- 15881668 TI - Possible involvement of glutamic and/or aspartic acid residue(s) and requirement of mitochondrial integrity for the protective effect of creatine against inhibition of cardiac mitochondrial respiration by methylglyoxal. AB - We had previously shown that creatine exerted a protective effect against inhibition of cardiac mitochondrial respiration by methylglyoxal (SinhaRoy S, Biswas S, Ray M, Ray S. Biochem J 372: 661-669,2003). In the present study, we have investigated the mechanism of this protective effect by specific amino acid modifying reagent and by several compounds, which are structurally related to creatine. The results show that the compounds, which contain guanidine group such as arginine and guanidinopropionic acid, exert a protective effect, which is quantitatively similar to creatine. This result suggests the presence of carboxylic acid(s) such as glutamic and/or aspartic acid(s) in the creatine binding site, which has been further supported by experiments with N-ethyl-5 phenyl isoxazolium-3'-sulfonate a reagent known to modify these amino acids. Both polarographic and spectrophotometric assays were performed with NADH as respiratory substrate by using a) submitochondrial particles by sonication, b) freeze-thawed mitochondria and c) mitochondria permeabilized by alamethicin treatment. The results of these studies as compared to that of intact mitochondria indicate that structural integrity of mitochondria is essential for the protective effect of creatine. PMID- 15881669 TI - Molecular characterization of tumor associated antigen in mice exposed to a hepatocarcinogen. AB - The present investigation is aimed to identify and characterize tumor-associated antigen (TAA) in animals exposed to hepatocarcinogen. Swiss albino mice showed an enhanced expression of an approximately 58 kDa glycoprotein in liver cells upon exposure to a potential hepatocarcinogen N-nitrosodibutylamine (DBN). Carcinogenesis induction in mice was monitored by assays of y-glutamyl transpeptidase (GGT), acetylcholine esterase (AChE), glutathione-S-transferase (GST) activities and the level of glutathione (GSH) in liver. DBN treated animals showed cell distortion and extensive necrosis as observed by histological examination. The over-expressed TAA was purified by ion-exchange chromatography and further characterized by SDS-PAGE. The carbohydrate contents and glycan linkage to the polypeptide backbone was analyzed by using the DIG glycan differentiation and de-glycosylation kits. The glycoprotein has glycan chains that are N-linked via asparagines to the polypeptide backbone. It was also observed that the molecule is rich in sialic acid residues with a significantly high carbohydrate to protein ratio (> 2:1). The over-expressed high molecular weight glycoprotein TAA was found to be highly immunogenic and could eventually be used to induce immune response in order to counter tumor regression. PMID- 15881670 TI - Morphological and oxidative alterations on Sertoli cells cytoskeleton due to retinol-induced reactive oxygen species. AB - Retinol (vitamin A) is involved in several cellular processes, like cell division, differentiation, transformation and apoptosis. Although it has been shown that retinol is a limitant factor for all these processes, the precise mechanisms by which retinol acts are still unknown. In the present study we hypothesised that alterations in the cytoskeleton of Sertoli cells induced by retinol supplementation could indicate an adaptive maintenance of its functions, since it plays an important role in the transformation process that we observed. Previous results demonstrated that Sertoli cells treated with retinol showed an oxidative imbalance, that leads the cell to two phenotypes: apoptosis or transformation. Our group has identified characteristics of Sertoli cells transformed by retinol which results in normal cell functions modification. In the present study the actin filament fluorescence assay and the deformation coefficient showed a modification in the morphology induced by retinol. We also observed an oxidative alteration in isolated cytoskeleton proteins and did not show alterations when these proteins are analyzed by electrophoreses. Our results showed an increase in mitochondria superoxide production and a decrease in nitric oxide levels. All results were partially or completely reverted by co-treatment of the antioxidant Trolox. These findings suggest that the cytoskeleton components suffer individual alterations in different levels and that these alterations generate a global phenotype modification and that these processes are probably ROS dependent. We believe that the results from this study indicate an adaptation of the cytoskeleton to oxidative imbalance since there was not a loss of its function. PMID- 15881671 TI - Expression of lexA targeted ribozyme in Escherichia coli BL-21 (DE3) cells. AB - Coding sequences for a hammerhead ribozyme designed to cleave lexA mRNA in a targeted manner was cloned under phage T7 promoter and expressed in E. coli strain BL-21 (DE3) expressing T7 RNA polymerase under the control of IPTG inducible lac UV-5 promoter. Ribozyme expression in vivo was demonstrated by RNase protection assay. Also, total RNA extracted from these transformed cells following induction by IPTG, displays site-specific cleavage of labeled lexA RNA in an in vitro reaction. The result demonstrates the active ribozyme in extracts of cell transformed with a recombinant cassette and goes beyond the earlier demonstration of the stability of in vitro synthesized ribozyme in cell extracts. The observed rise in lexA mRNA rules out any role for protease activity or resulting fragments of lexA protein in de-repression of RNA. PMID- 15881672 TI - Effects of hibernation on multicatalytic proteinase complex in thirteen-lined ground squirrels, Spermophilus tridecemlineatus. AB - Multicatalytic proteinase complex (MCP) was studied in skeletal muscle of the hibernating ground squirrel, Spermophilus tridecemlineatus. MCP was partially purified using a S-400 gel filtration column and Centricon concentrating devices and assayed fluorometrically using three AMC-labeled substrates. K(m) and V(max) values were determined for each substrate with no significant differences between the enzyme from euthermic versus hibernating animals when assayed at 23 degrees C. However, properties of MCP from euthermic and hibernating ground squirrels were differentially affected by low assay temperature (8-10 degrees C) and also differed from the mouse enzyme, the data indicating that ground squirrel MCP is better suited for low temperature function. MCP preferentially degrades oxidatively-damaged proteins and quantification of protein carbonyl content showed that the level of oxidatively-damaged protein in skeletal muscle decreased by > 75% during hibernation suggesting a continuing role for the MCP in the torpid state. PMID- 15881673 TI - A Kruppel zinc finger of ZNF 146 interacts with the SUMO-1 conjugating enzyme UBC9 and is sumoylated in vivo. AB - The OZF (ZNF146) protein is a 33 kDa Kruppel protein, composed solely of 10 zinc finger motifs. It is overexpressed in the majority of pancreatic cancers and in more than 80% of colorectal cancers. We have identified OZF interacting factors with a yeast two-hybrid screen. Half of the positive clones characterized encoded UBC9, the E2 enzyme involved in the covalent conjugation of the small ubiquitin like modifier 1 (SUMO-1). SUMO-1 is a 17 kDa migrating protein that is conjugated to several proteins and has been reported to exhibit multiple effects, including modulation of protein stability, subcellular localization, and gene expression. In HeLa cells transfected with OZF and SUMO-1 expression vectors, immunoblot revealed a major band migrating at 50 kDa and a minor band at 67 kDa, corresponding to the attachment to OZF of one and two SUMO-1 proteins, respectively. The relative amount of the sumoylated proteins increased following transfection with a UBC9 expression vector. The presence of the sumoylated form in HeLa cells solely transfected by OZF indicates the physiological activity of the endogenous SUMO-1 conjugation pathway. Using deletion mutants, we showed that two SUMO-1 modification sites are located on the sixth zinc finger. Mutation of two lysine residues greatly reduced the amount of the sumoylated form of OZF though their surrounding sequences differ from the consensus sequence reported for most proteins modified by SUMO-1 conjugation. Despite the presence of the sixth zinc finger, an OZF mutant containing zinc fingers 1-6 was not modified by SUMO-1 and failed to interact with UBC9. Addition of zinc finger 7 restored SUMO 1 modification and UBC9 interaction and provides evidence that a region downstream of the target lysines is required for interaction with UBC9, in order to achieve SUMO-1 modification. This is the first report of in vivo conjugation of a SUMO-1 protein to a Kruppel zinc finger motif. PMID- 15881674 TI - Bigendothelin-1 (1-21) fragment during early sepsis modulates tau, p38-MAPK phosphorylation and nitric oxide synthase activation. AB - Earlier we have demonstrated that inhibition of endothelin biosynthesis ameliorates endotoxemia-induced inducible nitric oxide synthase (iNOS) activation and phosphorylation of p38-mitogen activated protein kinase (pp38-MAPK). Therefore, in the present study, we tested the hypothesis that activation of endothelin (ET)-1 biosynthesis using bigET-1 during early sepsis would upregulate iNOS and affect myocardial function in the rat. Male Sprague-Dawley rats (350-400 g) were anesthetised using Nembutal (50 mg/kg, i.p.) and jugular vein, tail artery (Mean arterial pressure, MAP) and right carotid arteries (advanced to left ventricle, LV) were cannulated. The rats were randomly divided into saline-, bigET-1- and C-terminal fragment of bigET-1 (bigET-1(22-38))-treated groups. Sepsis was induced using i.p. injection of cecal inoculum obtained from a donor rat (200 mg/kg in 5 ml 5% sterile dextrose water, D5W). Sham animals received an i.p. injection of D5W (5 ml/kg). MAP and LVP were recorded and cardiodynamic parameters were calculated at 0, 2, 6, 12 and 24 h post sham or sepsis-induction. A significant elevation in LV isovolumic relaxation rate constant (tau), LV end diastolic pressure (LVEDP) and rate pressure product (RPP) was observed in vehicle-treated septic group at 24 h. BigET-1 significantly increased concentration of LV ET-1 both in sham and septic groups. BigET-1 elevated tau and LVEDP both in sham and septic animals as early as 12 h which persisted through 24 h. However, bigET-1(22-38) elevated LVEDP in septic group at 24 h but not in sham group. BigET-1 accentuated the levels of plasma nitric oxide byproduct (NOx) levels in both sham and septic animals at 6, 12 and 24 h. Sepsis increased myocardial iNOS at 24 h. BigET-1 significantly upregulated expression of myocardial iNOS and pp38-MAPK. The data suggest that increased substrate availability for ET-1 at the time of sepsis-induction contributes in diastolic dysfunction, iNOS activation and p38-MAPK phosphorylation. PMID- 15881675 TI - Natura non facit saltum. PMID- 15881676 TI - Theories of adaptation: what they do and don't say. AB - Theoretical work on adaptation has lagged behind experimental. But two classes of adaptation model have been partly explored. One is phenotypic and the other DNA sequence based. I briefly consider an example of each--Fisher's geometric model and Gillespie's mutational landscape model, respectively--reviewing recent results. Despite their fundamental differences, these models give rise to several strikingly similar results. I consider possible reasons for this congruence. I also emphasize what predictions do and, as important, do not follow from these models. PMID- 15881677 TI - Testing hypotheses regarding the genetics of adaptation. AB - Many of the hypotheses regarding the genetics of adaptation require that one know specific details about the genetic basis of complex traits, such as the number and effects of the loci involved. Developments in molecular biology have made it possible to create relatively dense maps of markers that can potentially be used to map genes underlying specific traits. However, there are a number of reasons to doubt that such mapping will provide the level of resolution necessary to specifically address many evolutionary questions. Moreover, evolutionary change is built upon the substitution of individual mutations, many of which may now be cosegregating in the same allele. In order for this developing area not to become a mirage that traps the efforts of an entire field, the genetic dissection of adaptive traits should be conducted within a strict hypothesis-testing framework and within systems that promise a reasonable chance of identifying the specific genetic changes of interest. Continuing advances in molecular technology may lead the way here, but some form of genetic testing is likely to be forever required. PMID- 15881678 TI - QTL mapping and the genetic basis of adaptation: recent developments. AB - Quantitative trait loci (QTL) mapping has been used in a number of evolutionary studies to study the genetic basis of adaptation by mapping individual QTL that explain the differences between differentiated populations and also estimating their effects and interaction in the mapping population. This analysis can provide clues about the evolutionary history of populations and causes of the population differentiation. QTL mapping analysis methods and associated computer programs provide us tools for such an inference on the genetic basis and architecture of quantitative trait variation in a mapping population. Current methods have the capability to separate and localize multiple QTL and estimate their effects and interaction on a quantitative trait. More recent methods have been targeted to provide a comprehensive inference on the overall genetic architecture of multiple traits in a number of environments. This development is important for evolutionary studies on the genetic basis of multiple trait variation, genotype by environment interaction, host-parasite interaction, and also microarray gene expression QTL analysis. PMID- 15881679 TI - Sex differences in recombination and mapping adaptations. AB - Since the raw material of marker based mapping is recombination, understanding how and why recombination rates evolve, and how we can use variation in these rates will ultimately help to improve map resolution. For example, using this variation could help in discriminating between linkage and pleiotropy when QTL for several traits co-locate. It might also be used to improve QTL mapping algorithms. The goals of this chapter are: (1) to highlight differences in recombination rates between the sexes, (2) describe why we might expect these differences, and (3) explore how sex difference in recombination can be used to improve resolution in QTL mapping. PMID- 15881680 TI - Genetics and adaptation in structured populations: sex ratio evolution in Silene vulgaris. AB - Theoretical models suggest that population structure can interact with frequency dependent selection to affect fitness in such a way that adaptation is dependent not only on the genotype of an individual and the genotypes with which it co occurs within populations (demes), but also the distribution of genotypes among populations. A canonical example is the evolution of altruistic behavior, where the costs and benefits of cooperation depend on the local frequency of other altruists, and can vary from one population to another. Here we review research on sex ratio evolution that we have conducted over the past several years on the gynodioecious herb Silene vulgaris in which we combine studies of negative frequency dependent fitness on female phenotypes with studies of the population structure of cytoplasmic genes affecting sex expression. This is presented as a contrast to a hypothetical example of selection on similar genotypes and phenotypes, but in the absence of population structure. Sex ratio evolution in Silene vulgaris provides one of the clearest examples of how selection occurs at multiple levels and how population structure, per se, can influence adaptive evolution. PMID- 15881681 TI - Studying genetics of adaptive variation in model organisms: flowering time variation in Arabidopsis lyrata. AB - Arabidopsis thaliana has emerged as a model organism for plant developmental genetics, but it is also now being widely used for population genetic studies. Outcrossing relatives of A. thaliana are likely to provide suitable additional or alternative species for studies of evolutionary and population genetics. We have examined patterns of adaptive flowering time variation in the outcrossing, perennial A. lyrata. In addition, we examine the distribution of variation at marker genes in populations form North America and Europe. The probability of flowering in this species differs between southern and northern populations. Northern populations are much less likely to flower in short than in long days. A significant daylength by region interaction shows that the northern and southern populations respond differently to the daylength. The timing of flowering also differs between populations, and is made shorter by long days, and in some populations, by vernalization. North American and European populations show consistent genetic differentiation over microsatellite and isozyme loci and alcohol dehydrogenase sequences. Thus, the patterns of variation are quite different from those in A. thaliana, where flowering time differences show little relationship to latitude of origin and the genealogical trees of accessions vary depending on the genomic region studied. The genetic architecture of adaptation can be compared in these species with different life histories. PMID- 15881682 TI - Ontogenetics of QTL: the genetic architecture of trichome density over time in Arabidopsis thaliana. AB - Although much is known about the molecular genetic basis of trichome development in Arabidopsis thaliana, less is known about the underlying genetic basis of continuous variation in a trait known to be of adaptive importance: trichome density. The density of leaf trichomes is known to be a major determinant of herbivore damage in natural populations of A. thaliana and herbivores are a significant selective force on genetic variation for trichome density. A number of developmental changes occur during ontogeny in A. thaliana, including changes in trichome density. I used multiple interval mapping (MIM) analysis to identify QTL responsible for trichome density on both juvenile leaves and adult leaves in replicate, independent trials and asked whether those QTL changed with ontogeny. In both juvenile and adult leaves, I detected a single major QTL on chromosome 2 that explained much of the genetic variance. Although additional QTL were detected, there were no consistent differences in the genetic architecture of trichome density measured on juvenile and adult leaves. The finding of a single QTL of major effect for a trait of known adaptive importance suggests that genes of major effect may play an important role in adaptation. PMID- 15881683 TI - Epistasis and genotype-environment interaction for quantitative trait loci affecting flowering time in Arabidopsis thaliana. AB - A major goal of evolutionary biology is to understand the genetic architecture of the complex quantitative traits that may lead to adaptations in natural populations. Of particular relevance is the evaluation of the frequency and magnitude of epistasis (gene-gene and gene-environment interaction) as it plays a controversial role in models of adaptation within and among populations. Here, we explore the genetic basis of flowering time in Arabidopsis thaliana using a series of quantitative trait loci (QTL) mapping experiments with two recombinant inbred line (RIL) mapping populations [Columbia (Col) x Landsberg erecta (Ler), Ler x Cape Verde Islands (Cvi)]. We focus on the response of RILs to a series of environmental conditions including drought stress, leaf damage, and apical damage. These data were explicitly evaluated for the presence of epistasis using Bayesian based multiple-QTL genome scans. Overall, we mapped fourteen QTL affecting flowering time. We detected two significant QTL-QTL interactions and several QTL-environment interactions for flowering time in the Ler x Cvi population. QTL-environment interactions were due to environmentally induced changes in the magnitude of QTL effects and their interactions across environments--we did not detect antagonistic pleiotropy. We found no evidence for QTL interactions in the Ler x Col population. We evaluate these results in the context of several other studies of flowering time in Arabidopsis thaliana and adaptive evolution in natural populations. PMID- 15881684 TI - Evolution in heterogeneous environments and the potential of maintenance of genetic variation in traits of adaptive significance. AB - The maintenance of genetic variation in traits of adaptive significance has been a major dilemma of evolutionary biology. Considering the pattern of increased genetic variation associated with environmental clines and heterogeneous environments, selection in heterogeneous environments has been proposed to facilitate the maintenance of genetic variation. Some models examining whether genetic variation can be maintained, in heterogeneous environments are reviewed. Genetic mechanisms that constrain evolution in quantitative genetic traits indicate that genetic variation can be maintained but when is not clear. Furthermore, no comprehensive models have been developed, likely due to the genetic and environmental complexity of this issue. Therefore, I have suggested two empirical approaches to provide insight for future theoretical and empirical research. Traditional path analysis has been a very powerful approach for understanding phenotypic selection. However, it requires substantial information on the biology of the study system to construct a causal model and alternatives. Exploratory path analysis is a data driven approach that uses the statistical relationships in the data to construct a set of models. For example, it can be used for understanding phenotypic selection in different environments, where there is no prior information to develop path models in the different environments. Data from Brassica rapa grown in different nutrients indicated that selection changed in the different environments. Experimental evolutionary studies will provide direct tests as to when genetic variation is maintained. PMID- 15881685 TI - The genetic basis of adaptation: lessons from concealing coloration in pocket mice. AB - Recent studies on the genetics of adaptive coat-color variation in pocket mice (Chaetodipus intermedius) are reviewed in the context of several on-going debates about the genetics of adaptation. Association mapping with candidate genes was used to identify mutations responsible for melanism in four different populations of C. intermedius. Here, I review four main results (i) a single gene, the melanocortin-1-receptor (Mc1r), appears to be responsible for most of the phenotypic variation in color in one population, the Pinacate site; (ii) four or fewer nucleotide changes at Mc1r appear to be responsible for the difference in receptor function; (iii) studies of migration-selection balance suggest that the selection coefficient associated with the dark Mc1r allele at the Pinacate site is large; and (iv) different (unknown) genes underlie the evolution of melanism on three other lava flows. These findings are discussed in light of the evolution of convergent phenotypes, the average size of phenotypic effects underlying adaptation, the evolution of dominance, and the distinction between adaptations caused by changes in gene dosage versus gene structure. PMID- 15881686 TI - The genetics of adaptation in Drosophila sechellia. AB - Drosophila sechellia is an island endemic of the Seychelles. After its geographic isolation on these islands, D. sechellia evolved into a host specialist on the fruit of Morinda citrifolia--a fruit often noxious and repulsive to Drosophila. Specialization on M. citrifolia required the evolution of a suite of adaptations, including resistance to and preference for some of the toxins found in this fruit. Several of these adaptive traits have been studied genetically. Here, I summarize what is known about the genetics of these traits and briefly describe the ecological and geographical context that shaped the evolution of these characters. The data from D. sechellia suggest that adaptations are not as genetically complex as historically thought, although almost all of the adaptations of D. sechellia involve several genes. PMID- 15881687 TI - Back to the future: genetic correlations, adaptation and speciation. AB - Genetic correlations can affect the course of phenotypic evolution. Although genetic correlations among traits are a common feature of quantitative genetic analyses, they have played a very minor role in recent linkage-map based analyses of the genetic architecture of quantitative traits. Here, we use our work on host associated races in pea aphids to illustrate how quantitative trait locus (QTL) mapping can be used to test specific hypotheses about how genetic correlations may facilitate ecological specialization and speciation. PMID- 15881689 TI - Hybridization as a source of evolutionary novelty: leaf shape in a Hawaiian composite. AB - Hybridization is increasingly recognized as a significant creative force in evolution. Interbreeding among species can lead to the creation of novel genotypes and morphologies that lead to adaptation. On the Hawaiian island of O'ahu, populations of two species of plants in the endemic genus Lipochaeta grow at similar elevations in the northern Wai'anae Mountains. These two species represent extremes of the phenotypic distribution of leaf shape: the leaves of Lipochaeta tenuifolia individuals are compound and highly dissected while leaves of L. tenuis are simple. Based primarily on leaf shape morphology, a putative hybrid population of Lipochaeta located at Pu'u Kawiwi was identified. Individuals in this population exhibit a range of leaf shapes intermediate in varying degrees between the leaf shapes of the putative parental species. We analyzed individuals from pure populations of L. tenuifolia, L. tenuis and the putative hybrids using 133 AFLP markers. Genetic analysis of these neutral markers provided support for the hybrid origin of this population. The correlation between genetic background and leaf morphology in the hybrids suggested that the genome of the parental species with simple leaves might have significantly contributed to the evolution of a novel, compound leaf morphology. PMID- 15881688 TI - Parallel genotypic adaptation: when evolution repeats itself. AB - Until recently, parallel genotypic adaptation was considered unlikely because phenotypic differences were thought to be controlled by many genes. There is increasing evidence, however, that phenotypic variation sometimes has a simple genetic basis and that parallel adaptation at the genotypic level may be more frequent than previously believed. Here, we review evidence for parallel genotypic adaptation derived from a survey of the experimental evolution, phylogenetic, and quantitative genetic literature. The most convincing evidence of parallel genotypic adaptation comes from artificial selection experiments involving microbial populations. In some experiments, up to half of the nucleotide substitutions found in independent lineages under uniform selection are the same. Phylogenetic studies provide a means for studying parallel genotypic adaptation in non-experimental systems, but conclusive evidence may be difficult to obtain because homoplasy can arise for other reasons. Nonetheless, phylogenetic approaches have provided evidence of parallel genotypic adaptation across all taxonomic levels, not just microbes. Quantitative genetic approaches also suggest parallel genotypic evolution across both closely and distantly related taxa, but it is important to note that this approach cannot distinguish between parallel changes at homologous loci versus convergent changes at closely linked non-homologous loci. The finding that parallel genotypic adaptation appears to be frequent and occurs at all taxonomic levels has important implications for phylogenetic and evolutionary studies. With respect to phylogenetic analyses, parallel genotypic changes, if common, may result in faulty estimates of phylogenetic relationships. From an evolutionary perspective, the occurrence of parallel genotypic adaptation provides increasing support for determinism in evolution and may provide a partial explanation for how species with low levels of gene flow are held together. PMID- 15881690 TI - Discovery and utilization of QTLs for insect resistance in soybean. AB - Insect resistance in soybean has been an objective in numerous breeding programs, but efforts to develop high yielding cultivars with insect resistance have been unsuccessful. Three Japanese plant introductions, PIs 171451, 227687 and 229358, have been the primary sources of insect resistance alleles, but a combination of quantitative inheritance of resistance and poor agronomic performance has hindered progress. Linkage drag caused by co-introgression of undesirable agronomic trait alleles linked to the resistance quantitative trait loci (QTLs) is a persistent problem. Molecular marker studies have helped to elucidate the numbers, effects and interactions of insect resistance QTLs in the Japanese PIs, and markers are now being used in breeding programs to facilitate transfer of resistance alleles while minimizing linkage drag. Molecular markers also make it possible to evaluate QTLs independently and together in different genetic backgrounds, and in combination with transgenes from Bacillus thuringiensis. PMID- 15881691 TI - Polyploidy, evolutionary opportunity, and crop adaptation. AB - The finding that even the smallest of plant genomes has incurred multiple genome wide chromatin duplication events, some of which may predate the origins of the angiosperms and therefore shape all of flowering plant biology, adds new importance to the molecular analysis of polyploidization/diploidization cycles and their phenotypic consequences. Early clues as to the possible phenotypic consequences of polyploidy derive from recent QTL mapping efforts in a number of diverse crop plants of recent and well-defined polyploid origins. A small sampling examples of the role(s) of polyploidy in conferring crop adaptation from human needs include examples of (1) dosage effects of multiple alleles in autopolyploids, and (2) 'intergenomic heterosis' conferring novel traits or transgressive levels of existing traits, associated with merging divergent genomes in a common allopolyploid nucleus. A particularly interesting manifestation of #2 is the evolution of complementary alleles at corresponding ('homoeologous') loci in divergent polyploid taxa derived from a common ancestor. Burgeoning genomic data for both botanical models and major crops offer new avenues for investigation of the molecular and phenotypic consequences of polyploidy, promising new insights into the role of this important process in the evolution of botanical diversity. PMID- 15881692 TI - Quantitative trait loci and the study of plant domestication. AB - Plant domestication ranks as one of the most important developments in human history, giving human populations the potential to harness unprecedented quantities of the earth's resources. But domestication has also played a more subtle historical role as the foundation of the modern study of evolution and adaptation. Until recently, however, researchers interested in domestication were limited to studying phenotypic changes or the genetics of simple Mendelian traits, when often the characters of most interest--fruit size, yield, height, flowering time, etc.--are quantitative in nature. The goals of this paper are to review some of the recent work on the quantitative genetics of plant domestication, identify some of the common trends found in this literature, and offer some novel interpretations of the data that is currently available. PMID- 15881693 TI - Can ecology help genomics: the genome as ecosystem? AB - Ecologists study the rules that govern processes influencing the distribution and abundance of organisms, particularly with respect to the interactions of organisms with their biotic and abiotic environments. Over the past decades, using a combination of sophisticated mathematical models and rigorous experiments, ecologists have made considerable progress in understanding the complex web of interactions that constitute an ecosystem. The field of genomics runs on a path parallel to ecology. Like ecology, genomicists seek to understand how each gene in the genome interacts with every other gene and how each gene interacts with multiple, environmental factors. Gene networks connect genes as complex as the 'webs' that connect the species in an ecosystem. In fact, genes exist in an ecosystem we call the genome. The genome as ecosystem is more than a metaphor--it serves as the conceptual foundation for an interdisciplinary approach to the study of complex systems characteristic of both genomics and ecology. Through the infusion of genomics into ecology and ecology into genomics both fields will gain fresh insight into the outstanding major questions of their disciplines. PMID- 15881694 TI - [Long follow-up of breast cancer revealed by clustered microcalcifications without palpable mass]. AB - OBJECTIVES: In the past two decades, nonpalpable breast cancer incidence has greatly increased by routine screening mammography. Microcalcifications are the most frequent radiological feature. METHODS: Between 1964 and 1994, 789 breast cancers revealed by clustered microcalcifications without palpable mass were treated at Institut Curie. Patients who had a past history of breast cancer or a contralateral cancer were eliminated like those with lobular carcinoma in situ. The definitive treatment was decided on the margins of the initial excision and the presence of microcalcifications on postoperative mammograms. RESULTS: Mean follow-up of the patients was 77 months (9-358). The treatment was conservative in 49.5% and radical in 50.5%. Histologically, 43.5% of the lesions were intraductal carcinoma, 25.5% were microinvasive and 31% were infiltrating cancers. Five and ten years overall survival were 98% and 92.5% respectively, 50% of the patients died from their breast cancer. This survival was identical for intraductal, microinvasive and invasive carcinomas without lymph node metastasis, but was less favorable in patients with node metastasis. There were no survival differences for the two treatment modalities, although local recurrences were more frequent in the conservative one. The 5-year recurrence rate was 3.5% for intraductal subtype, 4.4% for microinvasive and 6% for invasive cancers. The difference was not statistically significant. CONCLUSION: Under the impact of mammographic screening, sizable proportions of breast cancer patients are being seen in a totally nonpalpable state with either a very small invasive, microinvasive or intraductal breast cancer. These lesions have an excellent prognosis, and can benefit from less aggressive treatments. In the case of diffuse microcalcifications, where a radical treatment is indicated, skin sparing mastectomy can be easily done followed by immediate reconstructive surgery. PMID- 15881695 TI - Glycopeptide and oxacillin activity against Staphylococcus aureus isolates at a tertiary care center in Lebanon. AB - BACKGROUND: Worldwide reports have noted the emergence of glycopeptide intermediate Staphylococcus aureus (GISA). However, data from Lebanon is lacking. METHODS: We conducted a study to determine the minimum inhibitory concentrations (MICs) for oxacillin (OXA), vancomycin (VAN) and teicoplanin (TEC) against consecutive clinical isolates of S. aureus at a tertiary care center in Lebanon. The categorization of isolates as susceptible, intermediately-resistant, or resistant was based on the OXA agar screen plate (OS) and the 1 microg OXA disk diffusion (OD) according to the National Committee for Clinical Laboratory Standards (NCCLS) guidelines. The MICs of OXA, VAN and TEC against these isolates were determined using the E-test. RESULTS: We tested 147 S. aureus isolates, 108 of which were methicillin-resistant (MRSA) and 39 methicillin-susceptible (MSSA). The rar ges of MICs (microg/ml) among MRSA and MSSA isolates respectively were : OXA 1.5- > or = 256 and 0.19-3; VAN 1-3 and 1.5-3; TEC 0.094-3 and 0.19-2. The MIC50/90 (microg/ml) for MRSA versus MSSA isolates were, respectively: 24/ > or = 256 vs. 1/2 for OXA; 2/3 vs. 2/2 for VAN; 1/1.5 vs. 0.75/1.5 for TEC. CONCLUSION: This data indicates that despite the presence of high rates of MRSA (around 38%), these strains remain fully susceptible to VAN and TEC. We did not detect any GISA at this tertiary care referral medical center in Lebanon. PMID- 15881696 TI - Dosage of intrathecal baclofen maintenance therapy in the spastic syndromes. AB - Twenty-three patients affected with severe spasticity as a result of several clinical pathologies were treated with intrathecal Baclofen and administered at a lumbar level by means of an implantable infusion system. The system allows programming of any drug dosage in a continuous infusion fashion in the subarachnoid space. The appropriate patients were selected using specific criteria and the patients were regularly reviewed regarding the level of the spasticity and functional capacity. The required daily doses were determined using a specific detailed system. The doses needed varied according to the pathologic or clinical diagnosis. The doses administered by the global method at the beginning of the treatment were 91.96 microg/day and 137.81 microg/day in the final phase. These doses were higher in patients with medullary traumatic lesion, diffuse cerebral lesion and spastic idiopathic paraparesis. Relatively mild side effects were produced by the therapeutic doses used. PMID- 15881697 TI - [The particularities of the adult Scheuermann's disease: study about 45 patients]. AB - OBJECTIVE: Our aim was to find out which factors favor the occurrence of pain in adult patients with Scheuermann's disease--a juvenile manifestation of osteochondrosis of the spine, to study the clinical and radiological signs, the role of rehabilitation and the means to avoid the recurrence of pain. PATIENTS AND METHODS: Descriptive and retrospective study about 45 adults treated by rehabilitation. A data sheet listed the patient's occupation, current practice of sport, antecedents (violent sport, traumatism, pain), presence of a family form, motive of consultation, clinical and paraclinical examinations and immediate and long-term results of rehabilitation. RESULTS: Average age 35 years (18 to 65), male predominance (1.8:1), occupational risk factors: 16%; only 24% practiced sports currently. ANTECEDENTS: sports 49%, spinal column traumatism 13%, dorsal and lumbar pain during adolescence 16%. Consultation motive: abnormal posture 4% and pain 96%. According to the examination: abnormal spinal column in 80% of cases. According to radiography, our patients were divided into 49% who were carriers of growth vertebral dystrophy and 51% who were carriers of the real Scheuermann's disease. The immediate results of rehabilitation were satisfactory, 75% of the results were very good and good. Between six months and four years, 70% of the results were very satisfactory. Exercises were observed in only 11% of the cases, and stopped after two months on average. CONCLUSION: Scheuermann's disease in adults is a different entity from that of the teenager for the major manifestation is pain and not aesthetic quality. The patient's occupation is rather sedentary; sport is beneficial. The functional rehabilitation is the basic treatment and recourse to surgery or dorso-lumbar braces is rare. PMID- 15881698 TI - [Gene therapy: actualities and prospects]. AB - Gene therapy revolutionizes medicine by treating the causes of disease rather than the symptoms. This type of medicine would involve a therapeutic gene administered and a delivery system adapted to the targeted pharmaceutical objective. Although originally considered only as a treatment for inherited single-gene defects, gene therapy has also found applications in acquired illness, such as cardiovascular diseases. Finally, gene therapy requires stringent evaluation to see that it lives up to its enormous potential. PMID- 15881699 TI - [Gene therapy, where does it stand today]. PMID- 15881700 TI - Current treatment options for fissure-in-ano. AB - BACKGROUND: Fissure-in-ano is a painful condition that affects a sizable majority of the population. Selecting a method of treating the condition that could achieve optimal clinical results and the least pain and inconvenience to the patient has always posed a challenge to the surgeons. This led to the innovation of a number of surgical and pharmacological methods that relax the anal muscle. While acute fissures could be managed with medical therapy alone, chronic fissures do need some form of manipulation to relieve internal sphincter spasm. MATERIALS AND METHODS: The present study discusses various techniques advocated for the treatment of acute and chronic fissure-in-ano. It also elaborates on the advantages and deficiencies of each. DISCUSSION: Despite the initial success with pharmacological agents in the treatment of patients with chronic anal fissures, a growing concern is developing about their use. Increasing incidence of adverse effects and decreasing long-term efficacy have been the major drawbacks. CONCLUSION: Surgery still remains the preferred option which should be offered to patients with relapse or therapeutic failure with prior pharmacological treatment. Nevertheless, the patient should be informed about the pros and cons of each mode of treatment with details of cure rates and possible complications. PMID- 15881701 TI - The role of high sensitivity C-reactive protein in coronary artery disease risk prediction: a review. PMID- 15881702 TI - Atypical and unusual atrial myxoma. AB - A successfully excised right atrial myxoma arising from the suprahepatic segment of the inferior vena cava associated with asplenia, cutaneous manifestations, and iron deficiency anemia is reported. This report documents the unusual origin of myxoma, along with asplenia. A thorough review of the literature revealed only one other patient with right atrial myxoma originating from the suprahepatic inferior vena cava. Nevertheless, the literature contains no report of any other patient with asplenia associated with atrial myxoma. PMID- 15881703 TI - [von Hippel-Lindau syndrome: molecular diagnosis of two Lebanese families and analysis of the genotype-phenotype correlation]. AB - The von Hippel-Lindau syndrome (VHL) is a dominantly transmitted hereditary disorder associating multisystemic tumors affecting mainly the central nervous system, the kidneys, the pancreas, as well as pheochromocytomas. Mutations of the tumor suppressor gene VHL on chromosome 3 are responsible for the disease. This article reports for the first time the study of two Lebanese VHL affected families, presenting particularly hemangioblastomas of the central nervous system. Two different mutations of the VHL gene, S65W and F76S, respectively identified in the two families, confirmed the clinical diagnosis of the patients. Molecular diagnosis was then performed for at risk members of these families. This article reveals the importance of molecular diagnosis for suspected patients and of presymptomatic diagnosis for at risk members, especially that a close follow-up of carriers allows an early detection of tumors and prevents the metastasis stage, the most common cause of death of these patients. PMID- 15881704 TI - Imaging findings of radiation enteritis. AB - In this report the authors describe the imaging features of subacute and chronic radiation enteritis in two patients. Although surgery remains the definitive treatment of complicated chronic radiation enteritis, the potential role of interventional techniques that can be used for management of poor surgical candidates is discussed. PMID- 15881705 TI - Avoiding virologic armageddon in Africa. PMID- 15881706 TI - Antiretroviral therapy in advanced HIV-1 infection. AB - Current recommendations state that antiretroviral therapy (ART) should be commenced before the onset of severe HIV-associated immune deficiency and the development of AIDS-defining infections or malignancies. However, many patients only present and are diagnosed with HIV infection when they already have advanced disease. The optimal treatment for patients with advanced HIV disease remains to be defined. Key management questions include whether the virological and immunological responses to ART are comparable to those seen in patients with less advanced disease; whether the efficacy of different antiretroviral (ARV) regimens differs in patients with advanced disease; and whether there is an increased risk of drug toxicity and the immune reconstitution inflammatory syndrome. PMID- 15881707 TI - Genital manifestations in women with AIDS. PMID- 15881708 TI - Rapid response team reduces cardiac and respiratory arrests. PMID- 15881709 TI - Flow management brings peace to NH hospital's maternity ward. AB - Early mornings used to be a turbulent time in the maternity ward at Elliot Hospital in Manchester, NH, Obstetricians were scheduling many induced and caesarian deliveries to begin first thing in the morning, but often there weren't enough beds to handle both the scheduled and unplanned deliveries. PMID- 15881710 TI - Investing in technology and employees makes NJ hospital a quality leader. AB - When a patient arrives at the emergency department of the Robert Wood Johnson University Hospital Hamilton, in Hamilton Township, N.J., a long wait for service is not in the cards. That's because the hospital guarantees patients they'll be seen by a nurse within 15 minutes and a physician within 30 minutes. PMID- 15881711 TI - Kentucky health system to publicly report more than 200 indicators. AB - In a major upgrade of its performance improvement effort, Norton Healthcare of Louisville, KY, has announced plans to measure and publicly report more than 200 indicators by this spring. PMID- 15881712 TI - Medication safety guidelines offer minimum and best practice standards. AB - Guidelines from Wisconsin pharmacists offer drug safety measures to all healthcare professionals in a variety of settings. The guidelines are distinguishable from other recently released drug safety suggestions from several healthcare and government organizations because they have two levels: minimum and best practice standards. PMID- 15881713 TI - [Antiphospholipid antibodies and pregnancy]. AB - The presence of antiphospholipid antibodies is associated with a clinical syndrome characterised by thrombocytopenia, arterial and venous thromboses and recurrent fetal loss. Among theses antibodies currently detectable in laboratory are the lupus anticoagulant and anticardiolipin antibody. The antigens for theses antibodies are phospholipid-binding-proteins (prothrombin and beta 2 glycoprotein I). The exact mechanism of action of antiphospholipid antibodies remains controversal. The pathogenis of pregnancy loss seems related to the increased incidence of placental infraction. In spite of the improvement in our knowledge on the pathogenic mechanisms of the antiphospholipid syndrome, the standard therapy is still based on anti-platelet or anticoagulant drugs, both for vascular and obstetrical problems. Larger prospective clinical studies are needed to validate therapeutic implications. PMID- 15881714 TI - [Alternative medicine and dermatology]. AB - Alternative medicine is more popular than ever before and Dermatology has not remained unaffected by this trend. The aim of this review was to summarise all surveys of dermatological patients regarding the usage of alternative medicine to assess the potential efficacy of alternative medicine and to understand the reasons for which patients use alternative therapies. Computarized literature search was performed in medline. The search terms were "alternative medicine", "skin" and the name of different alternative therapies. Numerous studies have been practiced and published but few randomised controlled trials have been carried out. Greater knowledge of alternative medicine and its philosophical background and practical uses should result in better care for our patients and stimulate clinical research that will allow further consideration of its appropriate use and better evaluation of its possible danger. PMID- 15881715 TI - [Endoscopic treatment of residual lithiasis. 661 cases]. AB - AIM: Endoscopic sphincterotomy is a well-established procedure for treating choledocolithiasis and particularly residual lithiasis. The aim of this study is to expose our experience with this method and to evaluate his safety and efficacy. METHODOLOGY: It is a retrospectively collected series of 661 patients with residual choledocolithiasis. RESULTS: Endoscopic retrograde cholangio pancreatographies were performed in 651 patients (98.5%). Standard sphincterotomy was realized in 633 patients (95%) and a pre-cut technique was done in 15 (2%). Bile duct clearance was achieved in 583 cases (91%). Overall, the complication rate of sphincterotomy was about 8.4%. CONCLUSION: Endoscopic sphincterotomy is a safe and effective procedure for patients with residual choledocolithiasis. PMID- 15881716 TI - [Infectious urgencies in an emergency medical service]. AB - The aim of this study is to evaluate the frequency of infectious urgencies and the quality of their management in an emergency medical service. During the period of study, 21737 consultants are recorded, from which 2011 for an infectious urgency (9,25%), pharyngitis with rhinitis excluded. 692 patients are hospitalized and 1074 receive ambulatory antibiotics. Therapeutic urgencies are the most frequent infectious urgencies (48,4%), followed by protection urgencies (46,3%), urgencies due to antibiotics (5,2%) and prevention urgencies (0,1%). The conduct is considered correct for 1379 cases (68,5%) and incorrect for 500 cases (25%). Main incorrect conducts are the absence of follow-up (81,6%) and the maladjusted antibiotherapy (11,8%). These results incite, mainly, to the improvement of the professional training of emergency physicians and their communication with their colleagues of the other sanitary structures. PMID- 15881717 TI - [The hydatid cyst of the lung in children: 54 cases]. AB - The purpose of this retrospective study is to clarify the clinical, radiological and evolutionary aspects of the hydatid cyst of the lung. Over a period of 9 years (January, 1983 - December, 2001), we brought together in the pediatric service B of Children's hospital of Tunis, 54 cases of hydatid cyst of the lung. The average age is of 7 years 3 months (extremes 2 and 14 years), and the sex- ratio of 1,16. 61% of the children are of rural origin. Bronchpulmonary infection is the most frequent circumstance of discovery (72 % of cases). hemoptysis is revealing in 37% of cases. Discovery is fortuitous in 7% of cases. The radiography of the chest is of a big diagnostic contribution. The radiological aspects obtained are: a homogeneous opaqueness (61%), a diverse opaqueness (26%), an image of lung abscess (22 %), an aspect of floating membrane (4 %), a pleural effusion (9%), and an opaqueness with growing gas (4%). Chest echography was contributory in 77% of cases. 74% of the children were operated without complications. The others were lost. PMID- 15881718 TI - [Diagnostic strategy and management of anemia: a consensus from a detailed study of 100 patients followed in internal medicine]. AB - Anemia is a frequent reason for seeking medical care in Internal Medicine. It is not a diagnosis, but rather a symptom that needs to be investigated in order to find an etiology. In a retrospective review of 100 cases of anemia, the cause was found in 86 % of cases. The etiologic profile was varied. Iron deficiency remained by far the most frequent cause seen in 47% of case. Other etiologies have been found such as neoplasm in 13 % of cases, chronic renal failure in 8% of cases and endocrinal cause in 5 % of cases. We recommend a diagnostic decision tree that provides a tool for rational and cost effective approach for anemia. PMID- 15881719 TI - [Evaluation of clinical response by hydroxyurea in 132 patients with major sickle cell anemia]. AB - Hydroxyurea is used since close to 10 years in the treatment of major sickle cell anemia. In African sub Sahara, very little study have been dedicated. Yet, this drug represents nowadays, an alternative tot the bone marrow transplantation in the treatment of this illness. This study permits to illustrate the beneficial effects of this drung on 132 patients affected by of major sickle cell anemia. It is about patients whose middle age is of 28 years (range 14 and 54 years), most frequently masculine sex, followed regularly in the service of haematology of CHUB. The treatment consisted to the administration of the hydroxyurea to the posology between 10 and 30 mg/kg per day. The essentially clinical assessment focused on the sickle cell anemia crises and the modifications of hematologic's parameters. The experimentation has been pursued beyond the 18th month among 108 patients. It takes out again of this survey disappearances of the vasculo occlusive crises among 103 patients on 132 (78.03%) during the first 18 months of the survey. Only 10 patients (7.57%) benefited blood transfusion during the period of the survey because of the infectious complications. The hydroxyurea led a meaningful and spontaneous increase of the haemoglobin rate (+2,54 g/dl) of the middle globular volume (+ 16,02 femtolitres) as well as of the rate of haemoglobin foetale on 65 estimated patients. The hydroxyurea didn't bring any objective improvement on the pre-existing deneneratives complications. The treatment has been interrupted precociously among 5 patients for cytopenia. This survey confirms to short and middle term the beneficial effects of the hydroxyurea on the major sickle cell anaemia. PMID- 15881720 TI - [Endogamy and consanguinity in Mauritania]. AB - We examined the types of marriage between husband and wife and their frequency as well as the intensity of consanguinity for the Mauritanian ethnic groups. Then we evaluated for all the population, the variations of each of these dimensions according to the dates of marriage. The analisys of a sample of 2413 couples obtained according to the quota method showed that the Mauritanian population presents a strong family endogamy of about 47.20%. The Wolofs and the Poulards present pourcentages of marriages between blood relations of about 74.47% and of 55.74% respectively. On the other hand, for the Maures, black Maures and Soninkes, the proportions of marriages between blood relations are of the order of 41.25%, 30.52% and 46.99% respectively. This same analysis has also shown that the Mauritanian ethnic groupes present high apparent coefficient of consanguinity. They are of a level of unions between unequal cousins (1 /32) for (1/64) for others. The evaluation of endogamy practices in function of marriage dates (from 1921 to 1991) shows and for all the population a decline tendency in the pourcentages of consanguine marriages, and at the level of a whole Mauritanian population. PMID- 15881721 TI - [Gaucher's disease. Substitutive enzymatic treatment in one case and a review of the literature]. AB - Gaucher's disease, rare, hereditary and potentially mortal affection is characterized by the reduced concentration of the glucocerebroside lipid within the macrophage lysosomes. We report the case of a young 2 years old patient treated by transfusion since he was 9 months because of chronic anemia. According the clinical examination, the general state of the patients was bad ith important delayed stanturoponderal growth, a cutaneomucous paller and enormous splenomegaly. The blood count formula showed anemia with major thrombopenia. The myelogram was poor and the osteomedullar biopsy showed the presence of Gaucher's cells. The diagnosis has been confirmed by enzymatic dosage (Leucocytar b glucosidase). The treatment of the patient has been substitutive enzymatic (inifucerase) with very favorable response. During Gaucher's disease, the enzymatic deficiency results in the pathologic accumulation of the substrate (glucocerebroside) in the lyososomes, this metabolic overloading may cause polyvisceral disease with spontaneous evolution ofter mortal. The recent discovery of a recombining glucocerebrosidase (imiglucerase) transformed the prognosis of this disease. PMID- 15881722 TI - [Mature cystic teratoma of the fallopian tube. A case report]. AB - Primary teratomas of the fallopian tube are extremely uncommon. To date, only about 50 cases have been reported in the literature. They occur usually in the fourth decade. There association with nulliparity and reduced parity has been noted. Many have been discovered incidentally and none has been diagnosed preoperatively. The majority are cystic, show great variation in size and are commonly located in the ampulla or the isthmus. They can be intraluminal, attached to the serous surface by a pedicule and rarely intramural. Histologically they are similar to teratoma of the ovary and elsewhere. We describe a mature cystic teratoma of the fallopian tube found incidentally in a 35-year-old woman operated for suspicion of ectopic tubal pregnancy. The feature of benign teratoma of the fallopian tube is discussed. PMID- 15881723 TI - [Blind hemivagina: diagnostic method and problems of treatment]. AB - Blind hemivagina with duplex uterus and agenesis of the homolateral urinary tract is a rare malformation. For this reason, diagnosis may be not found. We report one case of duplex uterus with blind hemivagina in a fourteen-year-old adolescent girl. From this case, clinical signs, principal diagnosis criteria, complications and treatment are discussed, transvaginal resection of the hemivaginal septus is the best method of treatment but resulting hymeneal rupture may constitute a real problem of treatment in some social environments. So we propose to evaluate endoscopic septotomy. PMID- 15881724 TI - [Adenocarcinoma at the site of ileo-anal anastomosis in Crohn's disease: report of a case]. AB - We reported a case of patient born in 1967 who has been operated on in 1987 on three years after the beginning of ulcerative colitis which became severe and resistant to the medical treatment. A total colectomy with an ileo-rectal anastomosis had been performed. The pathological diagnosis carried of the specimen was Ulcerative colitis Then a proctectomy, followed by ileo-anal anastomosis, was performed in 1993. After several episodes of pochitis and the appearance of intestinal lesions upstream the ileal pocket, the retained diagnosis was Crohn's colitis. Eight years after the ileo-anal anastomosis, the patient developed an adenocarcinoma in the ileal pocket. He has been operated on in 2002 and he had abdominoperineal resection, radiotherapy and chemotherapy. Re died in January 2003. The death was related to the recurrence of malignancy. Endoscopic controls with biopsies are mandatory doing to follow up dysplasia predictive of degeneration. PMID- 15881725 TI - [Multiple coronary artery-left ventricular fistulae. A case report]. AB - The authors report the case of a patient with multiple coronary artery-left ventricular fistulae discovered in coronary angiography for exertional angina. PMID- 15881726 TI - Endodontics and pulp biology: the impact clinically. AB - Endodontists in clinical practice are uniquely situated to lead the effort of transferring translational laboratory and clinical research to their practices. This paper reviews the investigations presently being conducted around the world. The work presented is an overview of the progress being made to ensure retention of teeth with various forms of pulpal and periradicular diseases. PMID- 15881727 TI - Biologic perspectives to support clinical choices in root canal treatment. AB - Some of the most keenly debated issues in endodontics have revolved around the where to end the root filling, as well as cleaning and shaping and obturation techniques. In some respects, original scientifically-based concepts have been abandoned in favour of clinical techniques that may provide aesthetically pleasing radiographs, but do not necessarily support tissue regeneration around the root apex following obturation. Recently advocated obturation techniques supporting the use of resin-based and bonded materials have not had the same extensive evaluation that gutta-percha, when in contact with the periradicular tissues, has had. Further studies are necessary so that evidence-based data can support the confident use of these innovative materials. PMID- 15881728 TI - Treatment planning dilemmas resulting from failed root canal cases. AB - Endodontics is one of the least forgiving branches of dentistry in terms of treatment success or failure. As a result, re-treatment has become a significant part of today's endodontic practice. In the past, it was felt that surgical treatment with root-end filling would result in the highest degree of success. It has been subsequently demonstrated that microorganisms remaining in the root canal system after treatment were the main aetiological factor in post-treatment disease. Hence, non-surgical orthograde re-treatment has become the treatment of choice. While most authors support this method of re-treatment, some feel the surgical approach is still the best, and others advocate a combination of treatments. While treatment planning may appear to be straightforward, there are many grey areas in actual practice. This paper attempts to show a number of situations where the treatment planning may not be black and white. PMID- 15881729 TI - Apically-extruded debris using the ProTaper system. AB - The purpose of this in vitro study was to determine the quantity of debris and irrigant extruded apically using the ProTaper system compared to ProFiles and K Flexofiles. Thirty-six mesio-buccal root canals of human mandibular molars were selected and divided into three groups of twelve canals. Two groups were instrumented with ProFiles and ProTapers according to the manufacturer's instructions. The other group was instrumented with K-Flexofiles using the step back technique. A standard amount of irrigant was used for each canal. Apically extruded debris and irrigant was collected in pre-weighed vials. The mean weight of extruded debris and irrigant for each group was statistically analysed using Student's t-test and one-way ANOVA. All instrumentation techniques produced extruded debris and irrigant. Although the mean amount of extrusion with the step back technique was higher than the two rotary systems, there was no significant difference between the three groups (p > 0.05). NiTi rotary systems were associated with less apical extrusion, but were not significantly better than hand file instrumentation. All techniques extruded debris. PMID- 15881730 TI - Relationship of patient complaints and signs to histopathologic diagnosis of pulpal condition. AB - The correlation between the histopathologic examination of pulp biopsy specimens and patients' complaints and signs was investigated. The sensitivity, specificity and reliability of each complaint and sign, and the characteristics of pain that are associated with treatable and untreatable pulp states is proposed. Pulp specimens were obtained from teeth that required endodontic treatment. Clinical data were recorded to identify each patient's complaints. The pulp specimens were processed and the histopathologic diagnoses were categorised and correlated with the patients' complaints. Of the 240 cases, 100 (41.7%) were diagnosed as atrophic pulp or pulposis; 4 (1.7%) as acute pulpitis; 64 (26.7%) as transitional stage; 56 (23.3%) as chronic pulpitis, and 16 (6.7%) as acute pulpitis superimposed on a chronic pulpitis. Results showed that previous pain (p < 0.05), spontaneous pain (p < 0.01), and prolonged pain on cold stimuli (p < 0.05), were significantly more frequent in the patients with chronic pulpitis compared to those with pulposis or transitional stage. We concluded that clinicians must consider the sensitivity and specificity of patient complaints and signs in order to perform a diagnosis based upon clinical evidence. PMID- 15881732 TI - Diagnosis and treatment planning are essential prior to commencing endodontic treatment: discuss this statement as it relates to clinical endodontic management. AB - Diagnosis and treatment planning are essential to the practice of endodontics. Diagnosis aims to determine whether pathological involvement of the dental pulp has or is occurring. Treatment planning meanwhile, involves appropriately selecting cases, determining how difficult the treatment may be to perform on a specific individual and sequencing treatment procedures to achieve a healthy and functional dentition. In endodontic management, this may involve establishing whether the tooth is restorable and periodontally sound, the patient is able to tolerate the treatments and the clinician has the skills to perform the required treatment procedures. Careful consideration of these issues must be given prior to commencing treatment. PMID- 15881733 TI - Becoming an endodontist in the United States. PMID- 15881734 TI - Emergency preparedness planning involves nurses. PMID- 15881735 TI - You do make a difference. PMID- 15881736 TI - Fighting for nursing's issues needs your financial support. PMID- 15881737 TI - Legislative platform issues briefing. PMID- 15881738 TI - MNSA has successful convention. PMID- 15881739 TI - Important legislation to oppose. PMID- 15881740 TI - [Aflatoxins in newly harvested corn in Panama]. AB - Mycotoxins are fungal metabolites that may contaminate foods and feeds, resulting at times in important disease in humans and animals. Thirty-six samples of nine varieties of newly harvested corn (4 samples each variety) were analyzed in search of aflatoxins by thin layer chromatography and high pressure liquid chromatography, and also cultured for the presence of Aspergillus flavus. Of the 36 samples studied, one was contaminated with 1290 ppb aflatoxin B1, which is 258x the concentration suggested by WHO, placed at 5 ppb in food for human consumption. Culture of the 36 samples of corn resulted in growth of 55 colonies of A. flavus from all but two (1 and 6) of the 9 varieties. Of the 55 colonies of A. flavus obtained, 15 (27.3%) were toxigenic. Comments are made on the public health implications of these findings. PMID- 15881741 TI - [Laparoscopic splenectomy: the learning process]. AB - This is a report of our initial experience with laparoscopic splenectomy in children. From August 1997 to March 1999, 19 patients underwent laparoscopic splenectomy for various hematologic disorders. The ages ranged from 3 to 13 years. A four port technique was used to perform the procedure. All 19 procedures were completed successfully laparoscopically. The average surgical blood loss was 60 cc. The average operative time was 138 min and the length of stay in the hospital 2.5 days. One patient required a reintervention because of postoperative bleeding. We discuss the learning curve in our laparoscopic splenectomy cases. PMID- 15881742 TI - [Management of complications of hemodialysis vascular accesses]. AB - Trombosis of the vascular access is the most frecuent complication among patients with chronic renal disease. From 1995 on we performed the percutaneous recanalization of the access and considered the procedure as the best option. We analized the results in the two groups of patients treated at different periods and emphasized the importance of the early diagnosis of the thrombosis in the last group. The importance of the information given by the radiologist to the surgeon pre and postoperatively is also stressed. PMID- 15881743 TI - [Helicobacter pylori antibiotic sensitivity by microdilution]. AB - The gastric pathogen Helicobacter pylori has been recognized as the major aetiologic agent of chronic gastritis and peptic ulcers and also a risk factor for gastric cancer; eradication of H pylori prevents peptic ulcer recurrence and may also decrease the prevalence of gastric cancer in high risk populations around the world. Currently the only accepted indication for treatment is ulcer disease and maltosa, infected with Helicobacter pilory. However treatment is difficult and easily develops resistance. The elaboration of an antibiotic profile is recommended after a treatment failure. There is a lack of information in developing countries so the aim of this work was to determine the antibiotic profile of 51 strains isolated from patients gastric biopsies attended at Hospital San Juan de Dios in Costa Rica, using egg yolk broth and finding a resistance of 63.0% to metronidazole with a breakpoint of 8.0 microg/ml and 20.0% resistance to tetracycline (MIC1.0 microg/ml), 6.0% to clarithromicyn with a MIC of 0.125 microg/ml. There was no resistance to amoxicilin (MIC 0.015 microg/ml). The microdilution technique is very laborious, but highly reproducible with results accordingly to previous work, and we recommended it for the designing of therapeutical scheme. PMID- 15881744 TI - [Clinical use of new antidepressive agents with dual action (literature review)]. AB - The effective pharmacological treatment of depression started in 1958 with the introduction of iproniazide and imipramine. New agents quickly followed with more specific actions and a safer side effect profile. Very recently, fourth generation antidepressants with dual action have been introduced. These new agents pose a challenging dilemma. Is it better to develop drugs ever more selective towards specific monoamine receptor subpopulations, or drugs that act upon several monoamines in a more focused way? The priority seems to be the investigation of the interactions of the various monoaminergic systems. This paper reviews the clinical use of the new antidepressants that implement the notion of dual action as an important element for efficacy combined with receptor specific action as a basis for tolerability. PMID- 15881745 TI - [Sjogren syndrome associated with renal tubular acidosis type I]. AB - Primary Sjogren's Syndrome complicated with a renal tubular acidosis type 1 and hypocalcemic paralysis, as the principal clinical manifestation, is uncommon. Although the initial manifestations of the nephropathy are not well understood, it is believed that the invasion of mononuclear cells and the high level of circulating antibodies, play an important role in the pathogenesis of the disease. We present a patient with hypocalcemic paralysis as an initial manifestation of a latent Sjogren's disease. The glandular biopsy was normal, suggesting a mayor participation of an immunological humoral factor in the renal lesion. PMID- 15881746 TI - [Osteopenia and osteoporosis in postmenopausal women in Panama]. AB - We present the results of densitometry studies performed on 1031 females, from September 1996 to September 1997. The studies were subsidized by the Foundation for Metabolic and Bone Diseases and the pharmaceutical institutions. The results were classified according to the WHO recommendations, taking in consideration the age of the patients, in normal, with osteopenia and with osteoporosis. 821 (79.63%) of the patients suffered from osteopenia or osteoporosis and 807 (78.2%) were postmenopause: only 14% of them were normal and 86% with osteoporosis or osteopenia. PMID- 15881747 TI - [Sandimmun vs Neoral, comparison between two types of cyclosporine in patients with kidney transplantation]. AB - We present a comparative trial between Sandimmun, the original cyclosporine presentation, and a modified one, called Neoral, in microemulsion, to increase the blood absortion and stability of the product. We built for each product, an absorption kinetic curve and we applied it to three patients with stable renal function, using the same dosage that was used for ciclosporine. We observed a greater absorption and stability in the blood levels of Neoral. We analyzed hundred blood samples in 25 patients and we found a higher level of this product; this lead us to make possible that in 72% of the patients we could diminish the daily dosis in 5 to 10%. The patient's renal function was stable and no complications or secondary effects were present with Neoral. PMID- 15881748 TI - Slit-ventricle syndrome. PMID- 15881749 TI - Neurofiberscope-guided management of slit-ventricle syndrome due to shunt placement. AB - OBJECT: The purpose of this study was to evaluate an original neurofiberscope guided strategy for the management of slit-ventricle syndrome that occurs after shunt placement. METHODS: Between 1995 and 2003 15 patients with slit-ventricle syndrome (mean age 14.2 years) underwent endoscopic third ventriculostomy (ETV) and shunt removal. During the initial surgical procedure a neurofiberscope with a small outer diameter was inserted along the shunt tube into the collapsed ventricle for endoscopically controlled removal of the ventricular catheter and evaluation of brain compliance. If the latter was sufficiently preserved, primary ETV and shunt removal were performed (four cases). If brain compliance seemed to be significantly reduced, endoscopically controlled replacement of the ventricular catheter and implantation of the Codman-Hakim programmable valve shunt device were performed (11 cases). In these patients, delayed ETV and shunt removal were performed later (mean period of 16.3 months). No medical or surgical complications occurred in any case. Follow up ranged from 6 to 84 months (mean 31.1 months; median 22 months). All patients became shunt independent and 13 became symptom free. Overall, the size of the ventricles returned to normal in five cases, became slightly dilated in nine, and moderately dilated in one. CONCLUSIONS: Neurofiberscope-guided treatment of slit-ventricle syndrome involving shunt removal and ETV appears to be beneficial; all patients in this series were symptom free and shunt independent at the end of follow up. PMID- 15881750 TI - Spinal lesion level in spina bifida: a source of neural and cognitive heterogeneity. AB - OBJECT: The aim of this study was to evaluate whether the level of a spinal lesion is associated with variations in anomalous brain development and neurobehavioral outcomes in children suffering from the meningomyelocele form of spina bifida and hydrocephalus (SBM-H). METHODS: Two hundred sixty-eight children with SBM-H were divided into upper (T-12 and above; 82 patients) and lower (L-1 and below; 186 patients) lesion-level groups. Magnetic resonance images were qualitatively coded by radiologists and quantitatively segmented for cerebrum and cerebellum volumes. Psychometric assessments of handedness, intelligence, academic skills, and adaptive behavior were compared between lesion-level groups and also used to determine the number of children who met research-based criteria for mental retardation, attention deficit hyperactivity disorder, and learning disabilities. The magnetic resonance images obtained in children with upper-level spinal lesions demonstrated more qualitative abnormalities in the midbrain and tectum, pons, and splenium, although not in the cerebellum, compared with images obtained in children with lower-level spinal lesions. Upper-level lesions were also associated with reductions in cerebrum and cerebellum volumes, lower scores on measures of intelligence, academic skills, and adaptive behavior, and with a higher frequency of individuals meeting the criteria for mental retardation. Hispanic children (who were also more economically disadvantaged) were more likely to have upper-level lesions and poorer neurobehavioral outcomes, but lesion-level effects were generally independent of ethnicity. CONCLUSIONS: A higher level of spinal lesion in SBM-H is a marker for more severe anomalous brain development, which is in turn associated with poorer neurobehavioral outcomes in a wide variety of domains that determine levels of independent functioning for these children at home and school. PMID- 15881751 TI - Surgical management of temporal lobe tumor-related epilepsy in children. AB - OBJECT: Slow-growing, low-grade temporal lobe tumors are one of the most common causes of epilepsy in children. Although there are numerous consistent features in this patient group, consensus about the management and surgical approach is lacking. In this study the authors review the clinical, pathological, and radiological features as well as outcome data obtained in 29 pediatric patients with temporal lobe tumor-related epilepsy and discuss the surgical treatment strategies. METHODS: In patients who presented with intractable seizures secondary to mass lesions and underwent comprehensive epilepsy workup, the tumor was resected and the diagnosis confirmed by pathological examination. A minimum follow-up period of 16 months was required. Medical records were reviewed for details of seizure type and duration, electrophysiological data, imaging studies, operative notes, pathological examination reports, and follow-up data. The surgical approach was as follows. The lesionectomy with/without cortical resection was performed in all cases of lateral temporal tumors. Lesionectomy was performed with/without cortical resection in cases of basal temporal tumors if the mesial structures were radiologically normal. Mesial temporal tumors were excised, as were the remaining mesial temporal structures in the nondominant hemisphere; however, if the tumor was in the dominant hemisphere, lesionectomy was performed only if the remaining mesial structures were radiologically normal. Twenty-nine patients between 2 and 18 years of age were identified. Most tumors were located in the mesial temporal lobe. All patients underwent resection of the tumor with or without mesial and cortical structures. The most common pathological entity was dysembryoplastic neuroepithelial tumor. Sixty-nine percent of the patients remained seizure free (Engel Class I) and 14% experienced significant improvement (Engel Class II) after surgery. Outcome was better in the patients who underwent gross-total tumor resection. CONCLUSIONS: Mesially located low-grade neoplasms were the most frequently observed mass lesions in children with temporal tumor-related epilepsy in this series. Resection of the tumor with or without amygdalohippocampectomy provides a high rate of seizure-free outcome. It is the author's opinion that temporal lobe tumors should be managed based on the subgroups defined by their anatomical locations. If the tumor is located in or in proximity to eloquent cortex, we recommend functional magnetic resonance imaging and invasive monitoring techniques to map the eloquent cortex and epileptogenic zone, thereby tailoring the resection. PMID- 15881752 TI - Seizure outcome of lesionectomy in glioneuronal tumors associated with epilepsy in children. AB - OBJECT: Glioneuronal tumors (ganglioglioma, dysembryoplastic neuroepithelial tumors [DNTs]) are commonly associated with partial seizures. The optimal surgical treatment of such tumors, however, has not been fully established; it is still unclear whether lesionectomy itself can be used to control seizures or if epileptogenic areas adjacent to the tumor should also be removed. To address this uncertainty, the authors analyzed seizure outcome in a series of children with epileptogenic glioneuronal tumors that had been treated only by lesionectomy. METHODS: The authors retrospectively reviewed 15 children surgically treated for glioneuronal tumors associated with epilepsy. Patients ranged in age from 3 to 18 years (mean 12.6 years); there were 12 boys and three girls. The interval between onset of seizures and surgery ranged from 0.5 to 16 years (mean 6.1 years). Ten patients (66.6%) suffered complex partial seizures and five (33.3%) simple partial seizures. Seizure frequency varied from several per day to one per month. Nine tumors (60%) were temporal and six extratemporal; in all patients resection was limited to the tumor. The follow-up duration ranged from 1 to 11 years (mean 5.6 years). Gross-total removal was achieved in 13 patients and subtotal in two. The histological diagnosis was ganglioglioma in 11 cases and DNT in four. At last follow up 13 patients (86.6%) were Engel Class I, one was Engel Class II, and one was Engel Class III. CONCLUSIONS: The results of this study indicate that lesionectomy may provide good long-term seizure control in the majority of children with epileptogenic glioneuronal tumors. PMID- 15881753 TI - Detection of JC virus DNA sequences in brain tumors in pediatric patients. AB - OBJECT: The JC virus is a human neurotropic polyomavirus that causes progressive multifocal leukoencephalopathy and is closely related to simian virus 40. Several recent reports have indicated a possible association between the JC virus and the development of various human brain tumors. The authors examined the presence of JC virus DNA sequences in primary brain tumors in pediatric patients to evaluate the hypothesis that particular brain tumors can arise in the pediatric population as a consequence of infection with the JC virus. METHODS: Genomic DNA sequences were isolated from 62 brain tumors (32 medulloblastomas, 18 ependymomas, five choroid plexus papillomas, and seven pilocytic astrocytomas) and analyzed for the presence of JC virus DNA by Southern blot hybridization and direct sequencing. The JC virus DNA sequence was detected in five ependymomas and one choroid plexus papilloma. Immunohistochemical studies revealed nuclear expression of the large T antigen in a choroid plexus papilloma. None of the medulloblastomas or pilocytic astrocytomas contained JC virus DNA. CONCLUSIONS: The results of this study provide molecular evidence of the association between JC virus and the development of certain ependymomas and choroid plexus papillomas. PMID- 15881754 TI - Multicentric atypical teratoid/rhabdoid tumors occurring in the eye and fourth ventricle of an infant: case report. AB - Atypical teratoid/rhabdoid tumors (AT/RTs) are aggressive malignant tumors found in infants and young children. The tumor is characterized by the presence of a rhabdoid cell component in all cases, but the histological origin is still unclear. Recently, germline mutation of the hSNF5/INI1 gene has been reported in association with AT/RTs. The authors report a rare case of an intraocular AT/RT followed by a fourth ventricular tumor. The results of immunohistochemical studies of the surgical specimens revealed the presence of an AT/RT and from this finding the neural origin was inferred. A novel missense mutation of the hSNF5/INI1 gene was demonstrated by DNA analysis. High-dose chemotherapy with stem cell rescue was effective in treating this patient. The immunohistochemical relationship between rhabdoid cells and the neurogenic zone, which has not been described in AT/RTs, is of great interest in view of the nature of rhabdoid cells. PMID- 15881755 TI - Long-term control after resection and gamma knife surgery of an intracranial clear cell meningioma: case report. AB - Clear cell meningioma (CCM) is a rare variant of meningioma characterized by sheets of polygonal cells with clear cytoplasm, a feature attributable to its high glycogen content. Authors have described its propensity to recur and metastasize despite its benign pathological characteristics. Clinical response to radiation in these reports has varied. The authors present the case of a 7-year old girl with a large petroclival CCM who underwent a staged subtotal resection and subsequent gamma knife surgery (GKS). Initially, the residual tumor decreased in size, but 6 years later, it had regrown (9 mm in size). A second GKS treatment was performed and the mass completely regressed without further complication. The findings in this case suggest that GKS is a safe and effective adjunct for residual and recurrent CCM after resection. The delayed recurrence also emphasizes the importance of undertaking close follow-up examination after treating this potentially aggressive variant of meningioma. PMID- 15881756 TI - Incidental pediatric intraparenchymal xanthogranuloma: case report and review of the literature. AB - Juvenile xanthogranuloma (JXG) is a specialized form of non-Langerhans cell histiocyte proliferation that occurs in children. The majority of cases present as a solitary cutaneous lesion with a predilection for the head and neck region; however, isolated lesions occasionally have been identified in the central nervous system. The cutaneous forms of JXG usually follow a benign course. Other physicians have reported surgery as the first line of treatment in symptomatic patients with accessible lesions. Adjuvant therapies may be indicated for multicentric or surgically inaccessible lesions. The authors describe an unusual case of isolated intraparenchymal JXG in an asymptomatic child with no cutaneous manifestations and provide a review of the literature. PMID- 15881757 TI - Intradural mesh: an unusual cause of spinal cord tethering: case report. AB - A 13-year-old boy with a myelomeningocele experienced progressive foot deformity and lower-extremity pain while walking. Magnetic resonance imaging demonstrated a low-lying spinal cord with scarring near the site of a previous repair. During surgery, the terminal nerve roots were found to have scarred and adhered to a piece of metal mesh lying in the intradural space. The mesh had originally been placed to bridge a sacral ossification defect that was present at the initial closure of the child's myelomeningocele. PMID- 15881758 TI - Expansion of arachnoid cysts in children: report of two cases and review of the literature. AB - Arachnoid cysts are intracranial, space-occupying lesions that typically remain stable in size on serial imaging. The authors describe two cases of rapidly enlarging arachnoid cysts, including one located in the anterior fossa. In the first case a 7-month-old boy presented with increasing head circumference and a rapidly enlarging arachnoid cyst in the left middle fossa, which had been documented by serial imaging over the preceding 6 months. In the second case a 4 year-old girl presented with an arachnoid cyst compressing the right frontal lobe. The cyst had not been present on imaging studies performed during the perinatal period. In both cases, a craniotomy for open fenestration of the cyst was performed with successful resolution of the mass effect. Rare cases of expansion of arachnoid cysts have been reported in the literature. In this article the authors report the dramatic enlargement of two arachnoid cysts, including the first description of enlargement of an arachnoid cyst located in the anterior fossa. PMID- 15881759 TI - Craniopharyngioma and Cushing disease: case report. AB - Craniopharyngioma is a common sellar region tumor occurring in children. It usually manifests as endocrinological deficits such as short stature, delayed puberty, and obesity. Patients with craniopharyngioma commonly present with visual deficits and hydrocephalus. The authors present the case of a child who presented with short stature and clinical evidence of Cushing disease (CD) associated with a suprasellar tumor. The patient underwent insertion of an Ommaya reservoir into the tumor's cystic portion. High adrenocorticotropic hormone (ACTH) levels were demonstrated within the cyst's fluid and in the serum. After adequate decompression of the tumor, the patient underwent total resection. The tumor pathology was compatible with an adamantinomatous craniopharyngioma and immunohistochemical studies failed to show staining for ACTH. Panhypopituitarism developed postoperatively in the patient and he received hormone substitution therapy with final adequate height and normal-high weight. The neurosurgical implications of CD along with a possible mechanism for this patient's presentation are discussed in detail on the basis of the pertinent literature. PMID- 15881760 TI - Pineal region giant cell astrocytoma associated with tuberous sclerosis: case report. AB - Tuberous sclerosis complex is a genetic disorder characterized by the development of hamartomas in multiple organs including the brain, skin, eye, kidney, and heart. Neurological features include seizures and mental retardation. Cortical tubers and subependymal nodules are the characteristic intracranial lesions of tuberous sclerosis. Subependymal giant cell astrocytomas, typically located adjacent to the foramen of Monro, can enlarge and cause symptomatic ventricular obstruction. The authors describe the case of a 3-year-old boy with a history of tuberous sclerosis and retinal lesions who presented with an enlarging enhancing pineal region mass. Via an infratentorial supracerebellar approach, the mass was removed using both the operative microscope and a rigid neuroendoscope. Pathological examination showed a giant cell astrocytoma. To the authors' knowledge, this is the first reported case of tuberous sclerosis associated with a giant cell astrocytoma of the pineal region. Diagnostic considerations are discussed. PMID- 15881761 TI - Progressive cranial nerve palsy following shunt placement in an isolated fourth ventricle: case report. AB - Cranial nerve palsy is rarely seen after shunt placement in an isolated fourth ventricle. In the few reports of this complication, neuropathies are thought to be caused by catheter injury to the brainstem nuclei either during the initial cannulations or after shrinkage of the fourth ventricle. The authors treated a child who suffered from delayed, progressive palsies of the sixth, seventh, 10th, and 12th cranial nerves several weeks after undergoing ventriculoperitoneal shunt placement in the fourth ventricle. Magnetic resonance imaging revealed the catheter tip to be placed well away from the ventricular floor but the brainstem had severely shifted backward, suggesting that the pathogenesis of the neuropathies was traction on the affected cranial nerves. The authors postulated that the siphoning effect of the shunt caused rapid collapse of the fourth ventricle and while the cerebellar hemispheres were tented back by adhesions to the dura, the brainstem became the only mobile component in response to the suction forces. Neurological recovery occurred after surgical opening of the closed fourth ventricle and lysis of the basal cistern adhesions, which restored moderate ventricular volume and released the brainstem to its normal position. PMID- 15881762 TI - Neonatal cavernous carotid artery aneurysm: case report. AB - Neonatal intracranial aneurysms are rare. The authors report the case of a 4-week old girl who presented with left-eye ptosis and proptosis. Computerized tomography scanning and magnetic resonance imaging demonstrated a mass involving the left cavernous sinus and middle cranial fossa. Cerebral angiography revealed a large complex left cavernous carotid artery (CA) aneurysm. The patient underwent endovascular treatment in which detachable coils and N-butyl cyanoacrylate glue were used to achieve complete obliteration of the aneurysm. To the authors' knowledge, this is the first reported neonatal intracranial aneurysm originating from the cavernous CA and treated endovascularly. The authors review the literature on neonatal intracranial aneurysms. PMID- 15881763 TI - Ruptured superior cerebellar artery aneurysm in a child with bilateral fibromuscular hyperplasia of the renal arteries: case report and review of the literature. AB - The authors report the case of a pediatric patient with bilateral fibromuscular hyperplasia (FMH) of the renal arteries who sustained a subarachnoid hemorrhage (SAH) caused by the rupture of a superior cerebellar artery aneurysm. The aneurysm was successfully treated with microsurgical clip occlusion. The association between renal artery FMH and aneurysmal SAH is extremely rare and incompletely understood. There are six previously published cases in the English literature; only one case involved direct aneurysm treatment and none involved the posterior circulation. In addition to presenting this case, the authors discuss the complexities involved in treating these patients from a technical and conceptual standpoint. PMID- 15881764 TI - Transverse venous sinus stent placement as treatment for benign intracranial hypertension in a young male: case report and review of the literature. AB - Venous outlet obstruction has recently been reported to be a potentially treatable cause of benign intracranial hypertension (BIH). In the English language literature only 18 cases, all from the UK and Australia, involving the use of transverse sinus stenosis stent treatment for BIH have been reported; the youngest patient to receive treatment was a 17-year-old girl. The authors report the case of a 15-year-old boy who presented with headache, papilledema, decreased visual acuity, and diploplia who underwent successful unilateral transverse sinus stenosis stenting and experienced complete resolution of symptoms. PMID- 15881765 TI - Gel-based surgical preparation resulting in an operating room fire during a neurosurgical procedure: case report. AB - Despite general reports of fires in the operating room, those during neurosurgical procedures are rare. The most significant contributor to perioperative fires is excess oxygenation. The consideration of novel gel-based surgical preparation solutions as potential fuel sources should be included in the literature. Given the significant patient risk and legal ramifications of surgical fires, education regarding operating room fires and the potential for prevention and treatment is requisite. The authors describe the case of an 18 month-old girl who, while undergoing a resection of a right retroauricular scalp mass, suffered partial thickness bums as a result of the ignition of surgical preparation solution following the use of monopolar electrocautery. PMID- 15881766 TI - Transorbital penetrating brainstem injury in a child: case report. AB - The authors present the unusual case of a 9-year-old girl who sustained injury to her brainstem as a result of the orbital penetration of a metal projectile (nail) into the juxtamedullary region. This case and others reported in the literature associate this type of injury with relatively minor complications. Thorough imaging of the intracranial contents and surgical removal of the projectile is recommended. PMID- 15881767 TI - Dandy Walker. PMID- 15881768 TI - Stem cell therapy: a future treatment for heart failure? PMID- 15881769 TI - New drug treats form of age-related macular degeneration. Macugen inhibits progression of wet AMD and also may restore some vision. PMID- 15881770 TI - Rheumatoid arthritis patients have higher heart disease risk. Connection between the two is relatively new; inflammation may be the link. PMID- 15881771 TI - Statins may increase arthritis risk in women. PMID- 15881772 TI - Women don't take fractures seriously. PMID- 15881773 TI - The FLUKA code for space applications: recent developments. AB - The FLUKA Monte Carlo transport code is widely used for fundamental research, radioprotection and dosimetry, hybrid nuclear energy system and cosmic ray calculations. The validity of its physical models has been benchmarked against a variety of experimental data over a wide range of energies, ranging from accelerator data to cosmic ray showers in the earth atmosphere. The code is presently undergoing several developments in order to better fit the needs of space applications. The generation of particle spectra according to up-to-date cosmic ray data as well as the effect of the solar and geomagnetic modulation have been implemented and already successfully applied to a variety of problems. The implementation of suitable models for heavy ion nuclear interactions has reached an operational stage. At medium/high energy FLUKA is using the DPMJET model. The major task of incorporating heavy ion interactions from a few GeV/n down to the threshold for inelastic collisions is also progressing and promising results have been obtained using a modified version of the RQMD-2.4 code. This interim solution is now fully operational, while waiting for the development of new models based on the FLUKA hadron-nucleus interaction code, a newly developed QMD code, and the implementation of the Boltzmann master equation theory for low energy ion interactions. PMID- 15881774 TI - Role of shielding in modulating the effects of solar particle events: Monte Carlo calculation of absorbed dose and DNA complex lesions in different organs. AB - Distributions of absorbed dose and DNA clustered damage yields in various organs and tissues following the October 1989 solar particle event (SPE) were calculated by coupling the FLUKA Monte Carlo transport code with two anthropomorphic phantoms (a mathematical model and a voxel model), with the main aim of quantifying the role of the shielding features in modulating organ doses. The phantoms, which were assumed to be in deep space, were inserted into a shielding box of variable thickness and material and were irradiated with the proton spectra of the October 1989 event. Average numbers of DNA lesions per cell in different organs were calculated by adopting a technique already tested in previous works, consisting of integrating into "condensed-history" Monte Carlo transport codes--such as FLUKA--yields of radiobiological damage, either calculated with "event-by-event" track structure simulations, or taken from experimental works available in the literature. More specifically, the yields of "Complex Lesions" (or "CL", defined and calculated as a clustered DNA damage in a previous work) per unit dose and DNA mass (CL Gy-1 Da-1) due to the various beam components, including those derived from nuclear interactions with the shielding and the human body, were integrated in FLUKA. This provided spatial distributions of CL/cell yields in different organs, as well as distributions of absorbed doses. The contributions of primary protons and secondary hadrons were calculated separately, and the simulations were repeated for values of Al shielding thickness ranging between 1 and 20 g/cm2. Slight differences were found between the two phantom types. Skin and eye lenses were found to receive larger doses with respect to internal organs; however, shielding was more effective for skin and lenses. Secondary particles arising from nuclear interactions were found to have a minor role, although their relative contribution was found to be larger for the Complex Lesions than for the absorbed dose, due to their higher LET and thus higher biological effectiveness. PMID- 15881775 TI - [Preparation of solid lipid nanoparticles loaded with all-trans retinoic acid and their evaluation in vitro and in vivo]. AB - AIM: To prepare solid lipid nanoparticles (SLN) loaded with all trans retinoic acid using an ultrasonic technique with Compritol 888 ATO as matrix material, and investigate properties of nanoparticles in vitro and in vivo. METHODS: Ultrasonic technique was adopted to prepare solid lipid nanoparticles in an aqueous system using all-trans retinoic acid (ATRA) as a model drug. Physicochemical proterties of SLN were investigated in detail. Drug release from two sorts of ATRA-SLN was investigated using a dialysis bag method. Compared with ATRA solution, the in vivo pharmacokinetics of two sorts of ATRA-SLN after intravenous injection to rats were studied. RESULTS: Solid lipid nanoparticles loaded with all-trans retinoic acid was readily and quickly prepared by ultrasonic technique. The morphological investigation by Transmission Electron Microscopy (TEM) showed that the particles had round and uniform shapes. The mean diameters of them were (158 +/- 9) nm and (89 +/- 11) nm separately. The SLN dispersion was stable at 4 degrees C for more than one year. Drug loading was 3.3%, drug entrapment efficiency was more than 95%, the in vitro release was well in accordance with Weibull distribution. Compared with ATRA control solution, SLN could stay in the blood circulation for a longer time after intravenous injection. CONCLUSION: The ultrasonic technique was appropriate for the preparation of solid lipid nanoparticles. PMID- 15881776 TI - Detection of microlesions induced by heavy ions using liposomes filled with fluorescent dye. AB - In cells irradiation by heavy ions has been hypothesized to produce microlesions, regions of local damage. In cell membranes this damage is thought to manifest itself in the form of holes. The primary evidence for microlesions comes from morphological studies of cell membranes, but this evidence is still controversial, especially since holes also have been observed in membranes of normal, nonirradiated, cells. However, it is possible that damage not associated with histologically discernable disruptions may still occur. In order to resolve this issue, we developed a system for detecting microlesions based on liposomes filled with fluorescent dye. We hypothesized that if microlesions form in these liposomes as the result of irradiation, then the entrapped dye will leak out into the surrounding medium in a measurable way. Polypropylene vials containing suspensions of vesicles composed of either dipalmitoyl phosphatidylcholine, or a combination of egg phosphatidylcholine and cholesterol were irradiated at the Brookhaven National Laboratory using 56Fe ions at 1 GeV/amu. In several cases we obtained a significant loss of the entrapped dye above the background level. Our results suggest that holes may form in liposomes as the result of heavy ion irradiation, and that these holes are large enough to allow leakage of cell internal contents that are at least as large as a 1 nm diameter calcein molecule. PMID- 15881777 TI - Role of unmyelinated fibers in electroacupuncture cardiovascular responses. AB - The afferent fiber type responsible for the transmission of sensory neural traffic to the central nervous system during acupoint stimulation is uncertain. Several early studies evaluating compound action potentials have suggested that myelinated fibers contribute to the afferent input of the autonomic reflex adjustments during electroacupuncture (EA). Our more recent data, employing single unit recordings of somatic afferents, show that both myelinated and unmyelinated fibers are stimulated by EA, although more finely myelinated than unmyelinated fibers are activated by low frequency, low current stimulation. We hypothesized in this study that unmyelinated group VI fibers also contribute significantly to the inhibitory influence of EA on cardiovascular pressor responses. We found that neonatal capsaicin-treated rats depleted of substance P from primary afferents were insensitive to the inhibitory EA effect during gastric distention. Thus, EA at P5-P6 reduced gastric distention-induced pressor responses from 19+/-3 to 11+/-2 mmHg in eight untreated rats while capsaicin treated rats (n=9) were unresponsive to EA. Substance P containing neurons in dorsal root ganglion cells at Ti-T5 were significantly decreased in the capsaicin treated rats from 47+/-4 to 22+/-4 cells. Treated compared to untreated rats also demonstrated a significantly (P<0.03) reduced number of group IV fibers identified with single unit recording techniques. This study demonstrates that the inhibitory effect of EA at P5-P6 on cardiovascular autonomic excitatory reflexes involves unmyelinated group IV fibers of the median nerves. PMID- 15881778 TI - Cellular parameters for track structure modeling of radiation hazard in space. AB - Based on irradiation with 45 MeV/u N and B ions and with Co-60 gamma rays, cellular parameters of Katz's track structure model have been fitted for the survival of V79-379A Chinese hamster lung fibroblasts. Cellular parameters representing neoplastic transformations in C3H10T/1/2 cells after their irradiation with heavy ion beams, taken from earlier work, were also used to model the radiation hazard in deep space, following the system for evaluating, summing and reporting occupational exposures proposed in 1967 by a subcommittee of NCRP. We have performed model calculations of the number of transformations in surviving cells, after a given fluence of heavy charged particles of initial energy 500 MeV/u, penetrating thick layers of cells. We take the product of cell transformation and survival probabilities, calculated along the path lengths of charged particles using cellular survival and transformation parameters, to represent a quantity proportional to the "radiation risk factor" discussed in the NCRP document. The "synergistic" effect of simultaneous charged particle transfers is accounted for by the "track overlap" mode inherent in the model of Katz. PMID- 15881779 TI - Uncertainties in estimates of the risks of late effects from space radiation. AB - Methods used to project risks in low-Earth orbit are of questionable merit for exploration missions because of the limited radiobiology data and knowledge of galactic cosmic ray (GCR) heavy ions, which causes estimates of the risk of late effects to be highly uncertain. Risk projections involve a product of many biological and physical factors, each of which has a differential range of uncertainty due to lack of data and knowledge. Using the linear-additivity model for radiation risks, we use Monte-Carlo sampling from subjective uncertainty distributions in each factor to obtain an estimate of the overall uncertainty in risk projections. The resulting methodology is applied to several human space exploration mission scenarios including a deep space outpost and Mars missions of duration of 360, 660, and 1000 days. The major results are the quantification of the uncertainties in current risk estimates, the identification of factors that dominate risk projection uncertainties, and the development of a method to quantify candidate approaches to reduce uncertainties or mitigate risks. The large uncertainties in GCR risk projections lead to probability distributions of risk that mask any potential risk reduction using the "optimization" of shielding materials or configurations. In contrast, the design of shielding optimization approaches for solar particle events and trapped protons can be made at this time and promising technologies can be shown to have merit using our approach. The methods used also make it possible to express risk management objectives in terms of quantitative metrics, e.g., the number of days in space without exceeding a given risk level within well-defined confidence limits. PMID- 15881780 TI - Solar cosmic rays as a specific source of radiation risk during piloted space flight. AB - Solar cosmic rays present one of several radiation sources that are unique to space flight. Under ground conditions the exposure to individuals has a controlled form and radiation risk occurs as stochastic radiobiological effects. Existence of solar cosmic rays in space leads to a stochastic mode of radiation environment as a result of which any radiobiological consequences of exposure to solar cosmic rays during the flight will be probabilistic values. In this case, the hazard of deterministic effects should also be expressed in radiation risk values. The main deterministic effect under space conditions is radiation sickness. The best dosimetric functional for its analysis is the blood forming organs dose equivalent but not an effective dose. In addition, the repair processes in red bone marrow affect strongly on the manifestation of this pathology and they must be taken into account for radiation risk assessment. A method for taking into account the mentioned above peculiarities for the solar cosmic rays radiation risk assessment during the interplanetary flights is given in the report. It is shown that radiation risk of deterministic effects defined, as the death probability caused by radiation sickness due to acute solar cosmic rays exposure, can be comparable to risk of stochastic effects. Its value decreases strongly because of the fractional mode of exposure during the orbital movement of the spacecraft. On the contrary, during the interplanetary flight, radiation risk of deterministic effects increases significantly because of the residual component of the blood forming organs dose from previous solar proton events. The noted quality of radiation responses must be taken into account for estimating radiation hazard in space. PMID- 15881781 TI - Radiation analysis for manned missions to the Jupiter system. AB - An analysis for manned missions targeted to the Jovian system has been performed in the framework of the NASA RASC (Revolutionary Aerospace Systems Concepts) program on Human Exploration beyond Mars. The missions were targeted to the Jupiter satellite Callisto. The mission analysis has been divided into three main phases, namely the interplanetary cruise, the Jupiter orbital insertion, and the surface landing and exploration phases. The interplanetary phase is based on departure from the Earth-Moon L1 point. Interplanetary trajectories based on the use of different propulsion systems have been considered, with resulting overall cruise phase duration varying between two and five years. The Jupiter-approach and the orbital insertion trajectories are considered in detail, with the spacecraft crossing the Jupiter radiation belts and staying around the landing target. In the surface exploration phase the stay on the Callisto surface is considered. The satellite surface composition has been modeled based on the most recent results from the GALILEO spacecraft. In the transport computations the surface backscattering has been duly taken into account. Particle transport has been performed with the HZETRN heavy ion code for hadrons and with an in-house developed transport code for electrons and bremsstrahlung photons. The obtained doses have been compared to dose exposure limits. PMID- 15881782 TI - From mechanisms to risk estimation--bridging the chasm. AB - We have a considerable amount of work ahead of us to determine the importance of the wealth of new information emerging in the fields of sub-cellular, cellular and tissue biology in order to improve the estimation of radiation risk at low dose and protracted dose-rate. In this paper, we suggest that there is a need to develop models of the specific health effects of interest (e.g., carcinogenesis in specific tissues), which embody as much of the mechanistic (i.e., biological) information as is deemed necessary. Although it is not realistic to expect that every radiation-induced process should or could be included, we can hope that the major factors that shape the time dependence of evolution of damage can be identified and quantified to the point where reasonable estimations of risk can be made. Regarding carcinogenesis in particular, the structure of the model itself plays a role in determining the relative importance of various processes. We use a specific form of a multi-stage carcinogenic model to illustrate this point. We show in a review of the application of this model to lung cancer incidence and mortality in two exposed populations that for both high- and low LET radiation, there is evidence of an "inverse dose-rate" or protraction effect. This result could be of some considerable importance, because it would imply that risk from protracted exposure even to low-LET radiation might be greater than from acute exposure, an opinion not currently held in the radiation protection community. This model also allows prediction of the evolution of the risk over the lifetimes of the exposed individuals. One inference is that radiation-induced initiation (i.e., the first cellular carcinogenic event(s) occurring in normal tissue after the passage of the radiation) may not be the driving factor in the risk, but more important may be the effects of the radiation on already-initiated cells in the tissue. Although present throughout the length of the exposure, radiation-induced initiation appears to play a dominating role only very late in life, and only for those individuals who began their exposure early in life. These conclusions are very dependent, of course, on the hypotheses embodied in the initiation-promotion-conversion paradigm of carcinogenesis. We suggest that recently identified processes, such as the "bystander effect", might affect initiation, promotion, and malignant conversion in different ways. Finally, the manner in which the quality of radiation affects these processes must be understood in the context of the mixed high- and low-LET radiations that are found in the space environment. Important directions in critical experiment definition are suggested, including a renewed emphasis on well-designed animal experiments over extended periods of time. PMID- 15881783 TI - Austrian dose measurements onboard space station MIR and the International Space Station--overview and comparison. AB - The Atominstitute of the Austrian Universities has conducted various space research missions in the last 12 years in cooperation with the Institute for Biomedical Problems in Moscow. They dealt with the exact determination of the radiation hazards for cosmonauts and the development of precise measurement devices. Special emphasis will be laid on the last experiment on space station MIR the goal of which was the determination of the depth distribution of absorbed dose and dose equivalent in a water filled Phantom. The first results from dose measurements onboard the International Space Station (ISS) will also be discussed. The spherical Phantom with a diameter of 35 cm was developed at the Institute for Biomedical Problems and had 4 channels where dosimeters can be exposed in different depths. The exposure period covered the timeframe from May 1997 to February 1999. Thermoluminescent dosimeters (TLDs) were exposed inside the Phantom, either parallel or perpendicular to the hull of the spacecraft. For the evaluation of the linear energy transfer (LET), the high temperature ratio (HTR) method was applied. Based on this method a mean quality factor and, subsequently, the dose equivalent is calculated according to the Q(LET infinity) relationship proposed in ICRP 26. An increased contribution of neutrons could be detected inside the Phantom. However the total dose equivalent did not increase over the depth of the Phantom. As the first Austrian measurements on the ISS dosimeter packages were exposed for 248 days, starting in February 2001 at six different locations onboard the ISS. The Austrian dosimeter sets for this first exposure on the ISS contained five different kinds of passive thermoluminescent dosimeters. First results showed a position dependent absorbed dose rate at the ISS. PMID- 15881784 TI - Recent developments in employee benefits law. AB - The first part of this article highlights important judicial developments involving employee benefits and the Employee Retirement Income Security Act of 1974 ("ERISA"), as amended, during the latter part of 2003 and the first part of 2004, including the most significant U.S. Supreme Court and federal circuit court decisions. The second part covers recent legislative and regulatory developments in employee benefits law. This article is not meant to be exhaustive, but discusses the more important developments during 2003-2004, with particular focus on issues of concern to the insurance industry. PMID- 15881785 TI - Dose equivalent, absorbed dose and charge spectrum investigation in low Earth orbit. AB - Particle intensity, dose equivalent and absorbed dose have been measured on board the space shuttle Endeavour during STS-108 in December 2001 by Dublin Institute for Advanced Studies (DIAS). The dose estimates are based on very accurate measurements of recoils produced in CR-39 by cosmic ray primary and secondary protons and heavier nuclei and by secondary neutrons. The corresponding LET spectra were used to determine dose equivalent and absorbed dose values. Estimates of the total flux of Z > or = 2 nuclei have been undertaken and a preliminary charge spectrum was measured. Some comparisons are made with preliminary data obtained on STS-105 (ISS Expedition) and other missions using CR 39 detectors. PMID- 15881786 TI - The features of radiation dose variations onboard ISS and Mir space station: comparative study. AB - The dynamics of the ISS-measured radiation dose variations since August 2000 is studied. Use is made of the data obtained with the R-16 instrument, which consists of two ionization chambers behind different shielding thicknesses. The doses recorded during solar energetic particle (SEP) events are compared with the data obtained also by R-16 on Mir space station. The SEP events in the solar maximum of the current cycle make a much smaller contribution to the radiation dose compared with the October 1989 event recorded on Mir space station. In the latter event, the proton intensity was peaking during a strong magnetic storm. The storm-time effect of solar proton geomagnetic cutoff decreases on dose variations is estimated. The dose variations on Mir space stations due to formation of a new radiation belt of high-energy protons and electrons during a sudden commencement of March 24, 1991 storm are also studied. It was for the first time throughout the ISS and Mir dose measurement period that the counting rates recorded by both R-16 channels on ISS in 2001-2002 were nearly the same during some time intervals. This effect may arise from the decreases of relativistic electron fluxes in the outer radiation belt. PMID- 15881787 TI - Absorbed dose of secondary neutrons from galactic cosmic rays inside the International Space Station. AB - In this paper, we present the results of Monte-Carlo simulations of the flux and energy spectra of neutrons generated as a result of galactic cosmic ray proton interactions with the material of International Space Station (ISS) inside Zvezda Service Module, the Airlock between Russian and USA segments and one of Russian Research Modules for a full configuration of ISS. Calculations were made for ISS orbit for the energy ranges <10 and >10 MeV for both maximum and minimum of solar activity. To test the accuracy of the calculations the same simulations were made for MIR orbital station and for CORONAS-I satellite and compared with the results of measurements. Calculated and measured fluxes are in reasonable agreement. PMID- 15881788 TI - Trapped antiprotons produced by cosmic rays in the Earth's magnetosphere. AB - The existence of significant fluxes of antiparticles in the Earth magnetosphere has been predicted on theoretical considerations in this article. These antiparticles (positrons or antiprotons) at several hundred kilometers of altitudes, we believe are not of direct extraterrestrial origin, but are the natural products of nuclear reactions of the high energy primary cosmic rays (CR) and trapped protons (TP) confined in the terrestrial radiation belt, with the constituents of terrestrial atmosphere. Extraterrestrial positrons and antiprotons born in nuclear reactions of the same CR particles passing through only 5-7 g/cm2 of interstellar matter, exhibit lower fluxes compared to the antiprotons born at hundreds of g/cm2 in the atmosphere, which when confined in the magnetic field of the Earth (in any other planet), get accumulated. We present the results of the computations of the antiproton fluxes at 10 MeV to several GeV energies due to CR particle interactions with the matter in the interstellar space, and also with the residual atmosphere at altitudes of approximately 1000 km over the Earth's surface. The estimates show that the magnetospheric antiproton fluxes are greater by two orders of magnitude compared to the extraterrestrial fluxes measured at energies <1-2 GeV. PMID- 15881789 TI - Survey of current situation in radiation belt modeling. AB - The study of Earth's radiation belts is one of the oldest subjects in space physics. Despite the tremendous progress made in the last four decades, we still lack a complete understanding of the radiation belts in terms of their configurations, dynamics, and detailed physical accounts of their sources and sinks. The static nature of early empirical trapped radiation models, for examples, the NASA AP-8 and AE-8 models, renders those models inappropriate for predicting short-term radiation belt behaviors associated with geomagnetic storms and substorms. Due to incomplete data coverage, these models are also inaccurate at low altitudes (e.g., <1000 km) where many robotic and human space flights occur. The availability of radiation data from modern space missions and advancement in physical modeling and data management techniques have now allowed the development of new empirical and physical radiation belt models. In this paper, we will review the status of modern radiation belt modeling. PMID- 15881790 TI - Problems and conception of ensuring radiation safety during Mars missions. AB - The Mars mission differs from near-Earth manned space flights by radiation environment and duration. The importance of effective using the weight of the spacecraft increases greatly because all the necessary things for the mission must be included in its starting weight. For this reason the development of optimal systems of radiation safety ensuring (RSES) acquires especial importance. It is the result of sharp change of radiation environment in the interplanetary space as compared to the one in the near-Earth orbits and significant increase of the interplanetary flight duration. The demand of a harder limitation of unfavorable factors effects should lead to radiation safety (RS) standards hardening. The main principles of ensuring the RS of the Mars mission (optimizing, radiation risk, ALARA) and the conception of RSES, developed on the basis of the described approach and the experience obtained during orbital flights are presented in the report. The problems that can impede the ensuring of the crew members' RS are also given here. PMID- 15881791 TI - Performance of a simple Closed Aquatic Ecosystem (CAES) in space. AB - A simple Closed Aquatic Ecosystem (CAES) consisting of single-celled green algae (Chlorella pyrenoidosa, producer), a spiral snail (Bulinus australianus, consumer) and a data acquisition and control unit was flown on the Chinese Spacecraft SHENZHOU-II in January 2001 for 7 days. In order to study the effect of microgravity on the operation of CAES, a 1 g centrifuge reference group in space, a ground 1 g reference group and a ground 1 g centrifuge reference group (1.4 g group) were run concurrently. Real-time data about algae biomass (calculated from transmission light intensity), temperature, light and centrifugation of the CAES were logged at minute intervals. It was found that algae biomass of both the microgravity group and the ground 1 g-centrifuge reference group (1.4 g) fluctuated during the experiment, but the algae biomass of the 1 g centrifuge reference group in space and the ground 1 g reference group increased during the experiment. The results may be attributable to influences of microgravity and 1.4 g gravity on the algae and snails metabolisms. Microgravity is the main factor to affect the operation of CAES in space and the contribution of microgravity to the effect was also estimated. These data may be valuable for the establishment of a complex CELSS in the future. PMID- 15881792 TI - [Pathogenic substantiation and clinical use of the preparation Lycosol in comprehensive treatment of glossalgia]. PMID- 15881793 TI - Gain control by concerted changes in I(A) and I(H) conductances. AB - Stability of intrinsic electrical activity and modulation of input-output gain are both important for neuronal information processing. It is therefore of interest to define biologically plausible parameters that allow these two features to coexist. Recent experiments indicate that in some biological neurons, the stability of spontaneous firing can arise from coregulated expression of the electrophysiologically opposing I(A) and I(H) currents. Here, I show that such balanced changes in I(A) and I(H) dramatically alter the slope of the relationship between the firing rate and driving current in a Hodgkin-Huxley-type model neuron. Concerted changes in I(A) and I(H) can thus control neuronal gain while preserving intrinsic activity. PMID- 15881795 TI - Germ-line genetic enhancement and Rawlsian primary goods. AB - Genetic interventions raise a host of moral issues and, of its various species, germ-line genetic enhancement is the most morally contentious. This paper surveys various arguments against germ-line enhancement and attempts to demonstrate their inadequacies. A positive argument is advanced in favor of certain forms of germ line enhancements, which holds that they are morally permissible if and only if they augment Rawlsian primary goods, either directly or by facilitating their acquisition. PMID- 15881796 TI - Germ-line enhancement of humans and non-humans. AB - The current difference in attitude toward germ-line enhancement in humans and nonhumans is unjustified. Society should be more cautious in modifying the genes of nonhumans and more bold in thinking about modifying our own genome. I identify four classes of arguments pertaining to germ-line enhancement: safety arguments, justice arguments, trust arguments, and naturalness arguments. The first three types are compelling, but do not distinguish between human and nonhuman cases. The final class of argument would justify a distinction between human and nonhuman germ-line enhancement; however, this type of argument fails and, therefore, the discrepancy in attitude toward human and nonhuman germ-line enhancement is unjustified. PMID- 15881797 TI - Genetic enhancement: plan now to act later. AB - All three main articles in the issues of the Kennedy Institute of Ethics Journal endorse the view that genetic enhancement should be permitted, including human germ-line genetic enhancement. However, unregulated, wealth-based access to genetic enhancement in general, and germ-line enhancement in particular, would create intolerable risks for society. Although there are a number of practical problems raised by proposals to regulate or restrict access to genetic enhancement, which will make it difficult if not impossible to muster support for any effective restrictions until we begin to experience the societal problems that genetic enhancement will create, it is important to consider now what restrictions would be appropriate, how they would be imposed, and what changes would be needed in existing laws and institutions to facilitate them. Without this type of groundwork, there is no way society will be in a position to act in time. PMID- 15881798 TI - Genetic enhancement and the fate of the worse off. AB - When reflecting on arguments in the debate about genetic technologies, decision makers must try to be empathetic to those who are worse off. Disparities in health and health care in the U.S. pale when global facts are considered. Although U.S. citizens ought to be concerned about the worse off in the U.S., such concern ultimately must be balanced against the urgent imperative to address the plight of those in poor countries. It is a matter of fairness that care and concern be directed to those who are truly worse off in global terms. PMID- 15881799 TI - Justice in the genetically transformed society. AB - This paper explores some of the challenges raised by human genetic interventions for debates about distributive justice, focusing on the challenges that face prioritarian theories of justice and their relation to the argument advanced by Ronald Lindsay elsewhere in this issue of the Kennedy Institute of Ethics Journal. Also examined are the implications of germ-line genetic enhancements for intergenerational justice, and an argument is given against Fritz Allhoff's conclusion, found in this issue as well, that such enhancements are morally permissible if and only if they augment primary goods. PMID- 15881800 TI - Last word: imagining the future. AB - H.G. Wells warned, in 1895, not to allow economic injustices to become so acute that they ultimately transform human biology. Wells's warning is all the more pertinent today as society contemplates the use of biotechnologies to manipulate or "enhance" the human genome. PMID- 15881801 TI - Tetrazolo[1,5-a] quinoline derivatives as anti-inflammatory and antimicrobial agents [1]. AB - Tetrazolo[1,5-a]quinoline derivatives bearing in the 4-position various thiazolidinone 3a-c, 5a-c and 7a-c, thiazinone 8a,b, thiazoline 9a-d and thiadiazoline 10a,b moieties have been synthesized and evaluated for anti inflammatory activity and antimicrobial properties. The synthetic routes involved the reaction of tetrazolo[1,5-a]quinoline-4-carboxaldehyde 1 with amines and hydrazines to give the corresponding aniles 2a-c and hydrazones 4a-c respectively The latter compounds when treated with thioglycolic acid, furnished the corresponding thiazolidinone derivatives 3a-c and 5a-c. Moreover; thiosemicarbazone 6a-c derivatives were subsequently cyclized by various reagents giving rise the title compounds. Some of the products proved to possess potent anti-inflammatory and antimicrobial properties. PMID- 15881802 TI - Synthesis and biological evaluations of sulfa derivatives bearing heterocyclic moieties. AB - Some new sulfa derivatives bearing a heterocyclic moieties fural, pyrimidinone, thiazolidinone, benzimidazole and 1,2,4-triazinone and the related compounds 2-19 have been synthesized from treatment of sulfa drugs with thioisocyanate, acid chlorides, 3-chloro-1,2,4-triazines, aldehydes, esters and/or 2-methylbenzoxazole followed by ring closure reactions. Structures of the products have been deduced from their elemental analysis and spectral data. Significant antimicrobial activities were observed in vitro for some members of the series. Compounds 9b, 16 are highly active, while compounds 4b, 6d, 7,9a, 10 and 14 showing a moderate active towards gramme positive bacterium (b.subtilis). gramme negative bacterium (E. coli) and two fungi namely (A.nidulans & A.terreus). PMID- 15881803 TI - Spectrophotometric determination of captopril with DTNB reagent in pharmaceutical formulation. AB - A simple and sensitive spectrophotometeric method has been developed for the determination of captopril in its dosage form. The method is based on the reaction of the drug with DTNB reagent in pH 8 to produce a yellow coloured species measurable at 412 nm. The absorbance-concentration plot is linear over the range 1-10 x 10(-5) M with correlation coefficient of 0.997. The molar absorptivity and minimum delectability were 13553 and 3.2 x 10(-7) respectively. The proposed method was applied successfully for determination of captopril in its tablets form. The mean quantity and recovery were found to be 25.01+/-2% and 100.04+/-2% (each tablet contain 25 mg captopril). Quantification of the same tablets was determined by a standard method such as HPLC. The mean quantity of each tablet was found to be 25.07+/-1.22% by using HPLC method. It was not statistically, significant difference between Ellman's and HPLC method. This proposed method can be successfully applied to the determination of captopril in water and its dosage form and can use instead HPLC. PMID- 15881804 TI - Determination and correlation of the reversed-phase thin-layer chromatographic parameter (Rm) of a series of 3-(4-substituted-1-piperazinyl)-1-substituted-1 phenyl propanol derivatives with their LD50 values. AB - The experimental Rm values of four series of 3-(4-substituted-1- piperazinyl)-1 substituted-1-phenyl propanol derivatives, previously prepared for quantitative structure-activity relationship studies, were determined using reversed-phased thin-layer chromatography. The Rm values were determined for various concentrations of acetone in water (50, 60, 70, 80 and 90%), and the obtained correlation lines of Rm against proportion of acetone were extrapolated to 100% water The extrapolated Rm values of two of the four series of the 3-(4-benzyl-1 piperazinyl)-1-phenyl propanol derivatives were correlated with LD50 values obtained from 24-hour acute toxicity studies in albino mice. The extrapolated Rm values correlated parabolically with the LD50s. PMID- 15881805 TI - Assessment of pharmacy patients' compliance in Bulgaria (2001--2002). AB - The goal of this issue is to assess the level of compliance, respectively of non compliance among pharmacY patients between the years 2001 and 2002 in Sofia, Bulgaria. The authors study and emphasize on the factors that influence the non compliance of the patients and also try to find different ways to increase the rate of compliance in Bulgaria. Two types of standard individual questionnaires are applied for assessment of the state of compliance and non-compliance among the pharmacy patients. The obtained results show that the main reasons, influencing the non-adherence of the patients are: the high average price of the drugs, the overload daily regimen, inattention, distrust to the treatment and others. The drug form appears to be one of the main factors that cause non compliance, especially when the patients receive more than one drug in different drug forms few times a day and especially if elderly patients are addressed. Despite the presented comparatively high level of compliance among the patients (from 53% for 2001, to 82% for 2002), there are strategies to enhance compliance that have to be initiated. They incorporate communication tactics, patient's education, and proper dosage of the drugs and scheduling of the drug application. The results of the analysis of the received data show an increase in the level of compliance for the pharmacy patients in Bulgaria. For 2001 it was 53%, while for 2002 it has increased to 82%. This fact shows the influence of strategies for improvement of compliance. PMID- 15881806 TI - The influence of some aminoalkanolic xanthone derivatives on central nervous and cardiovascular systems in rodents. AB - A series of appropriate aminoalkanolic derivatives 2- or 4-methylxanthone was synthesized and evaluated for anticonvulsant activity in the maximal electroshock seizure (MES) and subcutaneous pentylenetetrazole seizure threshold (scPtz) assays, and for neurotoxicity (TOX). The most interesting result was the anticonvulsant activity of (R,S)-2N-(6-chloro-2-xanthonemethyl-2N-methylamino-1 propanol hydrochloride (II, which displayed anti-MES and anti-scPtz activity. Some of the obtained compounds (I - IV and V - VII) were also tested for their effect on the circulatory system (the effect on normal electrocardiogram, protection against adrenaline-, barium-, calcium- and/or strophanthine-induced arrhythmias, the effect on the arterial blood pressure and respiratory movements) and acute toxicity. PMID- 15881807 TI - Synthesis of new triazolopyrimidines and substituted pyrimidines. AB - Hydrazinopyrimidine 3 was prepared and transformed into 4, 5 by the reaction with Ar-Cl and POCl3, respectively. Also 3 transformed into 6, 7 by the reaction with ArCHO and FeCl3 respectively. Compound 8 was prepared and transformed into 9, 10, 11 by the reaction with DDC, ethyl bromoacetate and Sodium ethoxide respectively. PMID- 15881808 TI - Hyperhomocysteinemia is A risk marker for development of maternal pre-eclampsia. AB - The objective of the current study was to determine whether homocysteine elevations precede the development of pre-eclampsia, and to examine the relationship between the occurrence of pre-eclampsia and the degree of hyperhomocysteinemia, so as to find a new prognostic parameter for women with liable to develop pre-eclampsia. The study comprised 103 pregnant females chosen of those attending the Antenatal Care Unit at Benha University Hospital and accepted to donate blood samples at the 16th week of gestation. Women, who delivered at Benha University Hospital, were retrospectively allocated into two groups: Control group (Group C): comprised 64 (71.1%) parturient, who completed their full term pregnancy without the development of pre-eclampsia. Pre-eclampsia group (Group PEc): comprised 26 (28.9%) parturient who developed pre-eclampsia throughout their course of pregnancy but had completed their full term pregnancy. Through the present study, estimated fasting plasma tHcys levels were higher than the 90th percentile of the control group (> or = 5.1 ng/dl) in 6 (9.4%) women in group C and in 9 (34.6%) in group PEc. There was a significant (P<0.05) increase of fasting plasma tHcys levels in nullipara pre-eclamptic parturient as compared to multiparous control parturient. Also, a negative significant correlation was reported between parity and the fasting plasma tHcys level in pre-eclamptic parturient. The present results showed a significant increase of fasting plasma tHcys level in obese women with a positive significant correlation between fasting plasma tHcys level and BMI in PEc group. Thus, it can be concluded that hyperhomocysteinemia is an indirect risk factor for placental vasculopathy predating clinical pre-eclampsia, and can be used as a biomarker for identifying women at risk of complications and adverse pregnancy outcomes. PMID- 15881809 TI - Synthesis and antimicrobial properties of 3-sulfonyl-1,2,3,4-tetrahydropyrimido [4,5-d] pyrimidines. AB - The paper presents the synthesis of sulfonyl tetrahydropyrimido[4,5-d]pyrimidine derivatives as well as the antimicrobial activity of the obtained compounds. The investigations showed that the obtained sulfonyl derivatives of pyrimido[4,5 d]pyrimidines and especially the compounds with free amino group reveal interesting antibacterial and antifungal activity. PMID- 15881810 TI - Synthesis and anticonvulsant activity of some piperazine derivatives. AB - Various 1,4-substituted derivatives of piperazine (I-XII) were synthesized and evaluated for their anticonvulsant activity in the maximal electroshock (MES) and subcutaneous pentylenetetrazole (ScMet)--induced seizures and for neurotoxicity (TOX) in the rotorod test in mice and rats. The most promising compounds seem to be 1-[(2,4,6-trimethyl)-phenoxyethyl]-4-(2-hydroxyethyl)-piperazine dihydrochloride (II) and 1,4-bis-[(4-chloro-3-methyl)-phenoxyethyl]-piperazine dihydrochloride (X) which displayed anti-MES activity with their protective index (PI) higher than that for valproate II (rats), X(mice)). PMID- 15881811 TI - Photooxygenation of natural a-terpinene. AB - Photooxygenation reaction of a -terpinene (1) in the presence of tetraphenyl porphine (TPP) or Rose Bengal (RB) gave 2, 5-dihydroperoxy-1-isopropylene-4 methylene-cyclohexane (2), 3,6-dihydroperorxy-3-isopropyl-6-methyl-2,4 cyclohexadiene (3) and 1,2&4,5-diepoxy-1-isopropyl-4-methylcyclohexane (4). The epoxide 4 was also synthesised through the epoxidation reaction of a-terpinene (1) with m-chloroperbenzoic acid (mcpba). PMID- 15881812 TI - Antimicrobial activities of some amino derivatives of 5, 7-dibromo-2-methyl-8 benzoyloxyquinoline. AB - In an attempt to design and synthesize more potent quinoline-based chemotherapeutic agents structural modifications of 5, 7-Dibromo-2-methyl-8 benzoyloxyqinoline was carried out. The replacement of the bromine atoms with the requisite alkylamino compound gave four amino-derivatives viz: bis(dipropylamino) dipyrrolidino-, dipiperidino- and dipiperazino derivatives. The antimicrobial activities of these compounds were investigated against selected Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), Gram-negative bacteria (Escherichin coli, Pseudomonas aeruginosa and Klebsiella spp) and yeast (Candida albicans). All the compounds showed broad or significant antimicrobial activity, which varied from two to ninety times that of the parent compound. The dipyrrolidino derivative was the most effective against Gram-positive bacteria and yeast while the dipiperidino derivative was the most effective against Gram negative bacteria. No correlation has been established between the minimum inhibitory, (MIC) concentrations of the derivatives and the structural modifications. PMID- 15881813 TI - Bioequivalence study of sildenafil citrate tablets in healthy human volunteers. AB - Newly developed sildenafil citrate (SC), a selective inhibitor of cyclic guanosine monophosphate (c-GMP) specific phosphodiesterase type 5 (PDE 5) in the corpus cavernosum is used for the oral treatment of erectile dysfunction. A convenient, sensitive and simple method for the determination of sildenafil in human plasma is presented. The analytical technique was based on reversed-phase high-performance liquid chromatography coupled with UV detector set at 295 nm. Rofecoxib was used as internal standard (I.S). Liquid-liquid extraction using diethyl ether was performed to recover sildenafil and rofecoxib. The retention time of I.S and sildenafil were 5.5 minutes and 7.2 minutes respectively. The method was validated over a linear range of 10 to 1000 ng/ml from plasma. Separate stability study showed that sildenafil is stable under conditions of analysis. The extraction efficiency from plasma varied from 79.69% to 81.13 %. The minimum quantifiable concentration was set at 10 ng/ml. (%o CV<12.5%). The method was used for Bioequivalence Study of Two Brands of Sildenafil citrate 50 mg tablets in healthy human volunteers. All pharmacokinetic parameter were calculated along with statistical evaluation. PMID- 15881814 TI - Immobilization and release of etophylin from hydrogels, based on polyacrylic acid and macrodiisocyanates. AB - Hydrogels, based on polyacrylic acid and various macrodiisocyanates that contain polyethylene glycol, polypropylene glycol or polytetramethylene glycol in the main chain are synthesized. The obtained hydrogels are studied as possible polymer carriers of drug substances. Etophylin was applied as a model drug. It is exposed that the etophylin release rate from the hydrogel depends on the way of drug comprise in the polymer net, from its density from the chemical nature of the crosslinking agent and from the pH of the environment. The carried out studies let us to conclude that the obtained hydrogels are possible materials as drug carriers. PMID- 15881815 TI - Evaluation of crosslinked chitosan hydrogel beads as a carrier for prolonged delivery of diclofenac sodium: in vitro and in vivo studies. AB - Chitosan hydrogel beads containing diclofanac sodium were prepared using ionotropic gelation technique in which tripolyphosphate solution was used as a counterion. Chitosan molecular weight, tripolyphosphate concentration and crosslinking time were found to have an effect on the percentage of the drug loading. The loading efficiency of diclofenac sodium was very high (more than 90%). A longer-period of contact with the counterion ions decreased the efficiency of drug loading. The beads produced all had good spherical geometry. The beads showed a narrow size distribution in which 95% of the beads prepared were in the range of 2-3 mm. Comparison of release rate-time plots of dissolution data of marketed product with the newly developed dosage form indicated the ability of the later to sustain more diclofenac sodium release. The beads were evaluated for their bioavailability in six beagle dogs relative to the commercial enteric-coated Voltaren tablets. The in vivo availability study, reveled that the prepared beads filled in hard gelatin capsules had a 126.22% bioavailability relative to that of the commercial Voltaren tablets. The beads showed comparable pharmacokinetic parameters to that of the commercial tablets. The results suggested the possibility of producing a promising sustained drug delivery system for diclofenac sodium. PMID- 15881816 TI - Novel nicotinate esters of vasodilatation activity. AB - A variety of 2-substituted-4, 6-diaryl-3-pyridinecarboxylates 4 were obtained through aromatic nucleophilic substitution. reaction of secondary amines with 2 bromo-3-pyridinecarboxylate derivatives 3. The latters were obtained through bromination of 3-aryl-4-benzoyl-2-cyanobutyrates 2 in glacial acetic acid. However; reaction of primary aromatic amines with 2-bromopyridines 3 afforded 2 arylamino-3-pyridinecarboxylates 5 beside the unexpected 2-amino analogues 6. On the other hand, 3-hydroxy-1H-pyrazolo[3,4-b]pyridines 7 were isolated via reaction of 3 with hydrazine hydrate. Good to complete muscle relaxation of rabbit's jejunium, rat's uterus and rabbits aorta was observed during screening representative examples (3a, 4c, Sd, 5f and 6b) of the newly synthesized 3 pyridinecarboxylates indicating the vasodilatation and antihypertension activity for the tested compounds. PMID- 15881818 TI - The heartbreaking effects of the tsunami. PMID- 15881817 TI - Studies on the development of taste-masked suspension of chloroquine. AB - A non-bitter chloroquine suspension formulation for pediatric use was prepared in the form of an ion-pair of chloroquine with pamoic acid. Various parameters involved in the formulation of a stable and palatable suspension have been optimized. The suspension was characterized for particle size analysis, viscosity, physical and chemical stability and taste. Release of chloroquine from the ion-pair conducted as dissolution rate studies in simulated gastric media showed near to 100% release instantaneously. In-vivo bioavailability study conducted in albino rats indicated comparable bioavailability of chloroquine from the suspension with that of chloroquine phosphate syrup taken as standard. Stability study conducted over a period of 3 months showed the intactness of the ion-pair and the tasteless behavior of the suspension throughout the period of storage. PMID- 15881819 TI - Trends, challenges and status of the Ghanaian health care system. PMID- 15881820 TI - Health research, the MDGs and policy making in Mozambique. AB - Health research can play an important part in achieving the goals of improving health, economic development and reducing poverty, even though a review of the literature suggests that little attention is given to the subject. Mozambique as a developing country has scarce resources for health research and so its health research efforts need targeted at pressing concerns and be capable of influencing policy-making. A way forward is being developed so that position of policy-makers and researchers is clarified and co-ordination better organised. This will also involve a review of financing, formulation of agendas, the avoidance of wasted resources and improving management skills. PMID- 15881821 TI - Disaster preparedness in health care. AB - Natural disasters frequently cause major problems which affect a population's health and hinder a nation's socioeconomic development by draining its scarce financial resources in an effort to repair damages. For example, the tsunami disaster in the Indian Ocean is one of the worst natural disasters in modern times. Well over 200,000 people died and more than 1.5 million people lost their homes and often their livelihoods. Losses are estimated to total more than US dollar 7 billion. Private assets, including housing and business equipment, account for the largest share of the losses. In the largest countries, the impact on GDP is likely to be minimal, but the damage in the affected areas is extreme. Poor people were disproportionately affected. A country's health systems and public health infrastructure must be organised and ready to act in disaster situations as well as under normal conditions and must be cognisant of the type of measures to be taken in event of a disaster. These will differ according to the severity of the disaster's impact on the national health system. PMID- 15881822 TI - Economic factors influencing the implementation of community care for severely ill schizophrenic patients. AB - An increasing number of long-term schizophrenic patients are discharged from hospitals and taken care of in the community. This change in tendency has both a professional and economical side. The beneficial outcome of community care is well established. The aim of this paper is to appraise the economical implications and possibilities of community care compared to standard hospital care. The price elasticity for mental health services is higher than for other medical services. The demand for mental health care can not be directly compared with the demand for other types of care. This is due to lack of information on what is defined as good quality treatment in care and to define who is the demander of mental health care. Due to lack of defined demand and externalities encountered in the care for psychiatric patients, psychiatric treatment must be seen as a good that warrants government involvement in the financing and delivery of the service. The number in need of community care is estimated to be 12 per 100,000. To find the allocative efficiency in spending of resources on mental health care, it is important to look for the right balance between hospital and community care. There is evidence to assume that community care is more cost effective than hospital-based care. This does not apply to the most disabled schizophrenic patients where the costs are higher and outcome is the same. It is important to measure the costs incurred to family and friends when the total costs of community care are calculated and to find technically efficient production. The literature indicates that the physician/non-physician ratio has been too high. There are reports of dis-economies of scale, but economies of scope might be apparent. There are strong arguments in favour of state provision of psychiatric care for schizophrenic patients most in need. Otherwise the mental health care sector must be regulated with incentives that serve the need of the patients and encourage the most cost-effective treatment. Due to the risk of opportunism, specific assets and bounded rationality contracting involving clinical services should be avoided. PMID- 15881823 TI - Availability, safety and quality of blood for transfusion in the Americas. AB - OBJECTIVES: This article has two objectives: (1) to present for countries and territories of the Region of the Americas data on the number of blood donations, proportion of voluntary blood donors versus remunerated blood donors, coverage of screening for infectious agents, and separation of donated blood into its components and (2) to explore the relationships of those characteristics with economic and organisational factors in the countries and territories. METHODS: We carried out comparative analyses using population and health information gathered annually by the Pan American Health Organization (PAHO) from national health officials from the countries in the Americas, as well as economic information (gross national product (GNP) per capita) obtained from publications of the World Bank. RESULTS: There is a direct correlation between the availability of blood for transfusion and GNP per capita. Seven countries with a GNP per capita above US dollar 10,000 per year account for 38% of the Regional population but 68% of the Regional blood donations. Voluntary blood donation is more common in the countries with better blood availability. There is no association between GNP per capita and coverage of screening for infectious agents. Nevertheless, of the six countries with a GNP per capita below US dollar 1,000, only one of the six screens all units for human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B surface antigen (HBsAg). Countries with a higher proportion of voluntary blood donors tend to have lower prevalence rates of infectious markers. Separation of blood into its components is also more common in countries with higher blood donation rates. CONCLUSIONS: The availability, safety and quality of blood for transfusion in the Americas needs to be improved. As part of that effort, national policies and strategies must be put into place so that the resources already allocated for blood services are better utilised. PMID- 15881824 TI - What is eHealth?: a systematic review of published definitions. AB - CONTEXT: The term eHealth is widely used by many individuals, academic institutions, professional bodies and funding organisations. It has become an accepted neologism despite the lack of an agreed-upon clear or precise definition. We believe that communication among the many individuals and organisations that use the term could be improved by comprehensive data about the range of meanings encompassed by the term. OBJECTIVE: To report the results of a systematic review of published, suggested or proposed definitions of eHealth. DATA SOURCES: Using hte search query string 'eHealth' OR 'e-Health' OR 'electronic health', we searched the following databases: Medline and Premedline (1996-June 2004), EMBASE (1980-May 2004), International Pharmaceutical Abstracts (1970-May 2004), Web of Science (all years), Information Sciences Abstracts (1996 May 2004), Library Information Sciences Abstracts (1969-May 2004), and Wilson Business Abstracts (1982-March 2004), In addition, we searched dictionaries and an Internet search engine. STUDY SELECTION: We included any source published in either print format or on the Internet, available in English, and containing text that defines or attempts to define eHealth in explicit terms. Two of us independently reviewed titles and abstracts of citations identified in the bibliographic databases and Internet search, reaching consensus on relevance by discussion. DATA EXTRACTION: We retrieved relevant reports, articles, references, letters and websites containing definitions of eHealth. Two of us qualitatively analysed the definitions and coded them for content, emerging themes, patterns and novel ideas. DATA SYNTHESIS: The 51 unique definitions that we retrieved showed a wide range of themes, but no clear consensus about the meaning of the term eHealth. We identified two universal themes (health and technology) and six less general (commerce, activities, stakeholders, outcomes, place, and perspectives). CONCLUSIONS: The widespread use of the term eHealth suggests that it is an important concept, and that there is a tacit understanding of its meaning. This compendium of proposed definitions may improve communication among the many individuals and organisations that use the term. PMID- 15881825 TI - International medical ethics. PMID- 15881826 TI - Microbial metabolism of 2-chlorophenol, phenol and rho-cresol by Rhodococcus erythropolis M1 in co-culture with Pseudomonas fluorescens P1. AB - Chlorophenolic waste most often contains phenol and rho-cresol along with chlorophenols. A Rhodococcus erythropolis strain M1 was isolated with the ability to degrade 2-chlorophenol, phenol and p-cresol (100 mgl(-1), each) in 18, 24 and 20 h, respectively, with negligible lag. However, Rhodococcus sp. characterized by low growth rate, pose a threat to be outgrown by bacteria occurring in natural habitats. In the present study, interaction of R. erythropolis M1 with another isolated bacteria generally encountered in activated sludge for water treatment like Pseudomonas fluorescens P1 was studied. 2-chlorophenol, phenol and p-cresol were selected as the substrates for the study. Viable cell counts showed competitive interaction between the species on 2-chlorophenol and phenol. Specific growth rate of pure culture of R. erythropolis M1 was higher than P. fluorescens P1 on 2-chlorophenol. However, in mixed culture, P. fluorescens P1 showed higher growth rate. Degradation of phenol showed higher growth rate of R. erythropolis M1 both in pure and in mixed culture form. Degradation of p-cresol had shown similar counts for both populations indicating neutral type of interaction. This observation was substantiated by detecting the growth rate, where both cultures had similar growth rate in pure and in the mixed culture form. Rate of 2-chlorophenol degradation was higher when R. erythropolis M1 was used as the pure culture as compared to the degradation rates observed with the P. fluorescens P1 or with the mixed culture. However, in case of phenol and p cresol, degradation by the mixed culture had resulted in higher degradation rates as compared to the degradation of the substrates by both the axenic cultures. PMID- 15881827 TI - Identification and characterization of bacterial isolates from the Mir space station. AB - Twenty bacterial isolates (supplied by NASA) from the Mir space station water system were identified using Vitek GNI+ test card, API 20NE, and 16S rRNA gene sequencing. The identification of only one isolate agreed among the three techniques. The utility of the API 20NE and Vitek GNI+ test card approaches for identifying these isolates was Limited. Although 16S rRNA gene sequencing effectively identified many of the bacteria to the genus level, 74% of the isolates could not be identified to the species level. Isolates were also characterized based on motility and hydrophobicity. About 40% of the isolates were motile and four isolates were hydrophobic, suggesting that many of the bacteria have the potential to colonize surfaces and form biofilms. These findings demonstrate the difficulties in identifying bacteria from some environments to the species level and have implications for determining the risks of contamination in water systems of space shuttles and stations. PMID- 15881828 TI - Studies on a new marine streptomycete BT-408 producing polyketide antibiotic SBR 22 effective against methicillin resistant Staphylococcus aureus. AB - A new actinomycete strain designated as BT-408 producing polyketide antibiotic SBR-22 and showing antibacterial activity against methicillin resistant Staphylococcus aureus has been characterized and found to be a novel strain of Streptomyces psammoticus. Nutritional and cultural conditions for the production of antibiotic by this organism under shake-flask conditions have been optimized. Glucose and ammonium nitrate were found to be best carbon and nitrogen sources respectively for growth and antibiotic production. Similarly initial medium pH of 7.2, incubation temperature of 30 degrees C and incubation time of 96 h were found to be optimal. Optimization of medium and cultural conditions resulted in 1.82-fold increase in antibiotic yield. PMID- 15881829 TI - Isolation and characterization of diazotrophic growth promoting bacteria from rhizosphere of agricultural crops of Korea. AB - Free-living nitrogen fixing bacteria were isolated from rhizosphere of seven different plant namely sesame, maize, wheat, soybean, lettuce, pepper and rice grown in Chungbuk Province, Korea. Five isolates with nitrogenase activity above 150nmol(-1) mg(-1) protein were identified based on, phenotypic and 16S rDNA sequences analysis. The strains were identified as Stenotrophomonas maltophilia (PM-1, PM-26), Bacillus fusiformis (PM-5, PM-24) and Pseudomonas fluorescens (PM 13), respectively. All the isolates produced indole-3-acetic acid (IAA), in the presence of tryptophan, ranging from 100.4 microg ml(-1) (PM-13) to 255 microg ml(-1) (PM-24). The isolate PM-24 (Bacillus fusiformis) exhibiting highest nitrogenase activity (3677.81 nmol h(-1) mg(-1) protein) and IAA production (255microg ml(-1)) has a promising potential for developing as a plant growth promoting rhizobacteria. PMID- 15881830 TI - In vitro activity of three different antimicrobial agents against ESBL producing Escherichia coli and Klebsiella pneumoniae blood isolates. AB - Extended spectrum beta-lactamases (ESBLs) usually associated with multiple drug resistance, including beta-lactam and non-beta-lactam antibiotics. This resistance can cause Limitation in the choice of drugs appropriate for using in clinical practice, especially in life-threatening infections. In this study we aimed to investigate in vitro activity of meropenem, ciprofloxacine and amikacin against ESBL-producing and non-producing blood isolates of Escherichia coli and Klebsiella pneumoniae strains. Fifty-eight E. coli (21 ESBL-producing, 37 non ESBL producing) and 99 K. pneumoniae (54 ESBL-producing, 45 non-ESBL producing) strains were included in the study. The presence of ESBL was investigated by double disk synergy test and E-test methods. Antibiotic susceptibility test was done by microdilution method according to NCCLS guideline. In vitro susceptibilities of ESBL producing E. coli and K. pneumoniae strains were found as 100% for meropenem, 33.3% and 25.9% for ciprofloxacine, 94.5% and 83.3% for amikacin. It was observed that; meropenem was equally active agent in both ESBL producing and non-producing strains, and its activity was not affected by ESBL production. Whereas amikacin activity was minimally affected and ciprofloxacine activity was markedly decreased by ESBL production. In conclusion, meropenem seems to be better choice of antibiotic should be used for ESBL positive life threatening infections, because of remaining highest activity. PMID- 15881831 TI - Improvement of trypanocidal metabolites production by Aspergillus fumigatus using neural networks. AB - An optimization procedure using artificial neural networks was developed to determine the optimal combination of parameters, such as medium culture, initial pH, temperature and time of fermentation for maximal trypanocidal metabolites production by Aspergillus fumigatus. A data set of 81 experiments was carried out and an artificial neural network was trained to identify the optimal conditions for this process. Good correlation was obtained between the experimental and predicted values of lysis of the trypomastigote forms of Trypanosoma cruzi (r2 = 0.9990). The simulations of fermentation performance were undertaken on combinations of input variables and the highest level of activity against T. cruzi was obtained from the chloroform extract of the modified Jackson medium culture, initial pH of 6.0, incubated at 40 degrees C for 144 h. It displayed lysis of 95% of the trypomastigote forms of T. cruzi and the red blood cells remained normal. PMID- 15881832 TI - Changes in fatty acid composition in Pseudomonas putida and Pseudomonas stutzeri during naphthalene degradation. AB - The effects of naphthalene on the whole cell-derived fatty acid composition of Pseudomonas putida and Pseudomonas stutzeri during naphthalene degradation were investigated. These strains differed in their abilities to degrade naphthalene and in 1,2-catechol dioxygenase activities. The cells of both strains reacted to the addition of naphthalene with an increase in the saturated/unsaturated ratio. The dynamic changes comprised also alterations in the percentage of hydroxy, cyclopropane and branched fatty acids. Upon the exposure of naphthalene, new fatty acids were detected. PMID- 15881833 TI - Antibacterial, antifungal, and antiviral activities of the lipophylic extracts of Pistacia vera. AB - In the present study, antibacterial, antifungal, and antiviral properties of 15 lipohylic extracts obtained from different parts (leaf, branch, stem, kernel, shell skins, seeds) of Pistacia vera were screened against both standard and the isolated strains of Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Candida albicans and C. parapsilosis by microdilution method. Both Herpes simplex (DNA) and Parainfluenza viruses (RNA) were used for the determination of antiviral activity of the P. vera extracts by using Vero cell line. Ampicilline, ofloxocine, ketoconazole, fluconazole, acyclovir and oseltamivir were used as the control agents. The extracts showed little antibacterial activity between the range of 128-256 microg/ml concentrations whereas they had noticeable antifungal activity at the same concentrations. Kernel and seed extracts showed significant antiviral activity compared to the rest of the extracts as well as the controls. PMID- 15881834 TI - Alginate acetylation influences initial surface colonization by mucoid Pseudomonas aeruginosa. AB - Mucoid strains of Pseudomonas aeruginosa overproduce the exopolysaccharide alginate, which is substituted with O-acetyl groups. Under non-growing conditions in phosphate buffer, a mucoid clinical strain formed microcolonies on steel surfaces, while an acetylation-defective mutant was unable to form cell clusters. Enzymatic degradation of alginate by alginate lyase prevented microcolony formation of the mucoid parent strain. In a continuous-culture flow-cell system, using gluconate minimal medium, the mucoid strain with acetylated alginate formed microcolonies and grew into heterogenous biofilms, whereas the acetylation defective mutant produced a thinner and more homogeneous biofilm. A lowered viscosity of extracellular material from the acetylation-defective mutant indicated a weakening of exopolymer interactions by loss of acetyl groups. These results suggest that acetyl substituents are necessary for the function of high molecular-mass alginate to mediate cell aggregation into microcolonies in the early stages of biofilm development by mucoid P. aeruginosa, thereby determining the architecture of the mature biofilm. PMID- 15881835 TI - Organic acid exudation and pH changes by Gordonia sp. and Pseudomonas fluorescens grown with P adsorbed to goethite. AB - The actinomycete Gordonia sp. and the bacterium Pseudomonas fluorescens Pf-5 were grown in liquid media (pH 6.5) with phosphate adsorbed to the Fe-oxide/hydroxide goethite (Goe-P) and with soluble phosphate (0.1 mM or 1.0 mM P as KH2PO4). The two isolates showed distinct differences in their physiology. The pH of the medium was increased by Gordonia sp. by 1.1-1.7 units while it was decreased by P. fluorescens by 1.4-2.4 units. In all treatments the concentration of organic acids in the media with Gordonia sp. was up to 10 times lower (0.4-10.9 micromol L(-1)) than in media with P. fluorescens (33.4-84.4 micromol L(-1)). Gordonia sp. produced five different organic acids in varying amounts depending on P source and time. In contrast, P. fluorescens exuded mainly citrate and only small amounts of two to three other organic acids irrespective of P source or time. PMID- 15881836 TI - Chemical compositions and antimicrobial activities of four different Anatolian propolis samples. AB - Propolis means a gum that is gathered by bees from various plants. It is known for its biological properties, having antibacterial, antifungal and healing properties. The aims of this study were to evaluate the antimicrobial activity of four different Anatolian propolis samples on different groups of microorganisms including some oral pathogens and comparison between their chemical compositions. Ethanol extracts of propolis (EEP) were prepared from four different Anatolian propolis samples and examined whether EEP inhibit the growth of the test microorganisms or not. For the antimicrobial activity assays, minimum inhibitory concentrations (MIC) were determined by using macrodilution method. The MIC values of the most effective propolis (TB) were 2 microg/ml for Streptococcus sobrinus and Enterococcus faecalis, 4 microg/ml for Micrococcus luteus, Candida albicans and C. krusei, 8 microg/ml for Streptococcus mutans, Staphylococcus aureus, Staphylococcus epidermidis and Enterobacter aerogenes, 16 microg/ml for Escherichia coli and C. tropicalis and 32 microg/ml for Salmonella typhimurium and Pseudomonas aeruginosa. The chemical compositions of EEP's were determined by high-temperature high-resolution gas chromatography coupled to mass spectrometry. The main compounds of four Anatolian propolis samples were flavonoids such as pinocembrin, pinostropin, isalpinin, pinobanksin, quercetin, naringenin, galangine and chrysin. Although propolis samples were collected from different regions of Anatolia all showed significant antimicrobial activity against the Gram positive bacteria and yeasts. Propolis can prevent dental caries since it demonstrated significant antimicrobial activity against the microorganisms such as Streptococcus mutans, Streptococcus sobrinus and C. albicans, which involves in oral diseases. PMID- 15881837 TI - Comparison of microbial numbers in soils by using various culture media and temperatures. AB - The influence of different media and incubation temperatures on the quantification of microbial populations in sorghum, eucalyptus and forest soils was evaluated. Microbial growth was compared by using complex (tryptone soybean agar, TSA, casein-starch, CS, and Martin) and saline (Thorton, M3, Czapeck) media and incubation temperatures of 25 and 30 degrees C. Higher numbers of total bacterial and fungal colony-forming units (CFU) were observed in sorghum soils, and of spore-forming and Gram-negative bacteria in forest soils than other soils. Actinomycetes counts were highest in forest soil when using CS medium at 30 degrees C and in sorghum soil at 25 degrees C in M3 medium. Microorganism counts were dependent on the media and incubation temperatures. The counts at temperatures of 30 degrees C were significantly higher than at 25 degrees C. Microbial quantification was best when using TSA medium for total and spore forming bacteria, Thorton for Gram-negative bacteria, M3 for actinomycetes, and Martin for fungi. PMID- 15881838 TI - Pore formation, polymerization, hemolytic and leukotoxic effects of a new Enterobacter cloacae toxin neutralized by antiserum. AB - A new toxin of Enterobacter cloacae was purified and studied by SDS-PAGE electrophoresis with the purpose of investigating its ability to generate polymers and their molecular mass. Monomer of 13.3 kDa and structures of multimeric mass were detected. The toxin of 66 kDa was the most abundant form of toxin. This polymer and the monomer were selected to examine blood cells damage. Membrane pores caused by both toxin forms seemed to be of similar dimension (estimated in 3.6 nm by experiments with osmotic protectors) and were able to lyse erythrocytes and leukocytes. The results obtained indicate that polymerization and pore formation are involved in the molecular events that participate in the cytotoxic effects of E. cloacae toxin. Immunization of rabbits with 13.3kDa toxin generated antibody response capable of inhibiting oxidative stress as well as hemolytic and leukotoxic effects. Immunoblotting indicated that monomer and polymer reacted with antihemolysin serum. The importance of E. cloacae toxin "in vivo" was studied in animals by means of assays performed in peritoneum of rats, inoculated with the hemolytic strain (C1) and a non-hemolytic variant (C4). Both strains stimulated infiltration of leukocytes in peritoneum, but C1 caused cell death and lysis wheras assays with C4 maintained the viability of leukocytes even within 5 h after extraction of samples. PMID- 15881839 TI - Effect of environmental factors on production of lichenin, a chromosomally encoded bacteriocin-like compound produced by Bacillus licheniformis 26L-10/3RA. AB - Effect of environmental factors on production of lichenin, a bacteriocin-like compound produced by Bacillus licheniformis 26L-10/3RA isolated from buffalo rumen was studied. Lichenin represents the first anaerobiosis-specific expression of broad-spectrum antibacterial compound effective only under anaerobic conditions. Production of lichenin by B. licheniformis 26L-10/3RA was found to be very high at 39 degrees C in L-10 medium supplemented with 0.5% glucose and 20% (w/v) inert thermocol beads. Lichenin production was highest at pH 6.8 after 72 96h of incubation. Our study also indicated that Lichenin is not a plasmid-linked characteristic and is encoded by chromosomal DNA. Results obtained can be used in large-scale production of Lichenin for potential application in manipulating rumen function intended for improving productivity of the ruminants. PMID- 15881840 TI - [Towards better education for heart failure patients]. PMID- 15881841 TI - [Transient left ventricular apical ballooning or the tako-tsubo syndrome]. AB - Recently, Japanese authors have described a new clinical entity associating apical akidyskinesia and basal hyperkinesias without significant coronary artery disease under the name of the tako-tsubo syndrome. This syndrome is usually observed in elderly women and the clinical presentation is usually that of an acute coronary syndrome. The authors report 10 cases of patients investigated between June 2003 and August 2004. All patients were women with an average age of 66 +/- 15 years. Seven patients had chest pain on admission with, in three cases, signs of cardiac failure. In 8 patients, a causal factor was identified: emotional stress in 7 cases and anaphylactic shock in one case. All patients underwent coronary angiography and ventriculography which showed typical changes in left ventricular contractility without significant coronary disease. The peak CPK and troponin values were 222 +/- 115 UI/l and 3.32 +/- 1.50 microg/l, respectively. One patient died in cardiogenic shock. In the other cases, normalisation of wall motion was observed in the month following the onset of symptoms. One patient developed 3rd degree atrioventricular block and required implantation of a pacemaker. This clinical entity must be taken into consideration in elderly women with acute coronary syndromes. PMID- 15881842 TI - [Comparison between three-dimensional electro-anatomical mapping and conventional mapping in the ablation of atrial tachycardias]. AB - In atrial tachycardias, catheter ablation using conventional mapping system is associated with high immediate success and low recurrence. Three-dimensional electroanatomical mapping system combined to catheter ablation of atrial tachycardias has reached, in small uncontroled series, success rates of 100%. However, limited data are available about rates of recurrence or complication using this approach. In order to compare both mapping systems, we have conducted a study of 65 consecutive patients (36 women and 29 men) that underwent both electrophysiologic study and catheter ablation for suspected atrial tachycardias. Pre-existing heart disease was noted in 43%, hypertension in 32% and a history of atrial fibrillation of flutter in 52%. Catheter ablation guided by conventional mapping was undertaken in 44 patients and by three-dimensional electroanatomical mapping in 21. Successful ablation was performed in 68% of patients with conventional mapping and in 90% with three-dimensional electroanatomical mapping. No complication and recurrence were observed with the latter approach, while 5 patients had a recurrence and 2 had immediate complication with conventional mapping. Catheter ablation of atrial tachycardias combined with three-dimensional electroanatomical mapping appeared to be effective and safe, however, conventional mapping system still remains a reliable approach that must be considered as the first choice for atrial tachycardias ablation. PMID- 15881843 TI - [Automobile driving and implantable defibrillators]. AB - The consequences of implanting an automatic cardioverter defibrillator (ICD) on vehicle driving in France are poorly known. This retrospective study examined the behaviour at the wheel of ICD recipients who were recommended to abstain from driving for 3 to 6 months after device implantation. The study population included 98 patients (mean age = 59.5 +/- 14.8 years) followed for a mean of 24. +/- 23.9 months, who underwent ICD implant for ventricular tachycardia (65% of patients ventricular fibrillation (15%), syncope (8%), as part of a research protocol of myocardial cell transplantation 6%, or for primary prevention (5%). The underlying heart disease was ischemic in 59% of patients dilated cardiomyopathy in 11%,hypertrophic cardiomyopathy in 8%, valvular in 6%. Brugada syndrome in 4%, right ventricular arrhythmogenic cardiomyopathy in 2%, and miscellaneous disorders in 9% of patients. Five patients died without post mortem interrogation of the ICD. Only 28% of drivers remembered, and 13% observed, the recommended driving limitations. However, 45% (the oldest) claimed to drive prudently. During follow-up, 47% of patients received an ICD shock. Their mean it ventricular ejection fraction was 34 +/- 14%, versus 43 +/- 18% in patients who received no ICD therapy (p = 0.015). Syncope occurred in 16% who received ICD shocks. Shocks were delivered during driving in 6 patients, without consequent accident. Despite their non-observance of recommended driving limitations. ICD recipients suffered few traffic accidents. Legislation in France should reproduce the guidelines issued by European professional societies and enacted by the British laws. PMID- 15881844 TI - [Preliminary clinical experience with the Ventrica automatic distal anastomosis system in coronary surgery]. AB - The surgical treatment of coronary lesions is based on bypassing the anatomical lesions with autologous vascular grafts. This procedure has traditionally been "invasive", requiring a micro-surgery technique, institution of extra-corporeal circulation, as well as temporary cardiac arrest with a cardioplegic solution. Recently, an automatic distal anastomosis procedure has been developed (Ventrica, Medtronic Inc.), based on a magnetic coupling with two implanted intravascular magnets, allowing easy connection between the graft and the coronary artery. The immediately obvious advantages are the time saved, ease of use, reproducibility and reliability. The learning curve is fast. Furthermore, the use of this automatic process does not compromise a manual anastomosis in case of implantation failure. The immediate post-operative results, as well as angiography immediately and at 6 months are all satisfactory. This technique is applicable for multiple revascularisations, all types of autologous grafts, terminal or sequential bypasses, as well as nearly all types of coronaries. The contribution to beating heart, closed thorax coronary surgery seems equally promising. PMID- 15881845 TI - [Therapeutic education for cardiac failure patients: the I-care programme]. AB - Therapeutic education is becoming increasingly important in the management of chronic diseases including cardiac failure. The I-CARE programme consists of an evaluation of the role of therapeutic education in France, creating standardised tools and setting up training sessions for therapeutic education in the context of cardiac failure. Approximately two thirds of the French centres contacted perform therapeutic education with their available means. The lack of personnel, space, and training tools represent obstacles to the development of therapeutic education. The tools developed in the programme fall into 5 areas: diagnosis education, understanding the illness, diet, physical activity/daily life, and treatment. Training sessions were organised for the teams, consisting of at least one cardiologist and nurse. The I-CARE programme should allow the expansion of therapeutic education for cardiac failure and improve the multidisciplinary management of this disease which increasingly affects often elderly subjects. PMID- 15881846 TI - [Remodelling in atrial fibrillation]. AB - It is well known that atrial fibrillation (AF) tends to become permanent with time as illustrated by the fact that it becomes more difficult to maintain sinus rhythm when AF has been present for a long time. Atrial remodelling plays a part in this process and has been studied in experimental models. Atrial remodelling is defined as all the phenomena occurring during AF contributing to its maintenance. The persistence of AF induced by stimulation in the animal depends on the duration and the repetition of the atrial stimulation and it would appear that "AF induces AF". The tendency for AF to persist is associated with a shortening of the effective atrial refractory period with loss of its adaptation to the heart rate. The determining factor of both electrical and structural remodelling is the rapidity of the atrial rhythm of the AF itself. These phenomena lead to a type L calcium cellular overload as shown by its prevention or attenuation by the administration of verapamil, a calcium antagonist. Electrical remodelling is accompanied by a structural remodelling in the experimental model of persistent AF over several weeks. Samples from the two atria examined by electronic microscopy show mitochondrial changes, an accumulation of glycogen, a deficit in myofibrils, a redistribution of the nuclear chromatim and a reduction of sarcoplasmic reticulum with changes in protein structure. This structural remodelling is a reaction of adaptation similar to that observed in hibernating myocardium during ischaemia and aims to prolong cellular viability by decreasing atrial contractility. Another aspect of structural remodelling is the activation of fibroblasts with formation of fibrosis with resulting heterogeneity of the conduction tissue. There is also an increase in converting enzyme and angiotensin II concentrations. Irbesartan, an angiotensin II antagonist, reduced fibroblast growth. This has clinical applications as shown by the reduction in the recurrences of AF after cardioversion when given in association with amiodarone. Persistent AF leads to left atrial dilatation with abnormal atrial contractility. Further studies are necessary to determine the effect of atrial remodelling which might also act on the foci responsible for inducing the AF. Better understanding of atrial remodelling will contribute to the use of new pharmacological agents to prevent AF. PMID- 15881847 TI - [Safety of new anti-arrhythmic drugs]. AB - The majority of new antiarrhythmic drugs are still undergoing clinical trials and are not yet available on the French market. The studies of efficacy are mainly targeted on the treatment of atrial fibrillation. Intravenous ibutilide prolongs the duration of the action potential by stimulating sodium exchange during phase 2 of the action potential. Used for terminating episodes of atrial flutter and fibrillation, ibutilide has been shown to have a low proarrhythmic effect. Dofetilide is a pure I(Kr) current antagonist and is given orally. The molecule prolongs the duration of the atrial and ventricular action potentials. The amplitude of this effect is inversely related to the heart rate. No effect has been observed on the mortality rate in the post-infarct period. Adjusting the dosage with respect to renal function has reduced the occurrence of torsades de pointe from 4.8 to 2.9%. Azimilide is an I(Kr) and I(Ks) current blocker and its efficacy decreases at rapid heart rates. After oral administration, azimilide does not appear to have a deleterious effect in patients with a history of myocardial infarction and left ventricular dysfunction. The risk of torsades de pointe is less than 1%. Cases of neutropaenia have been reported. Dronedarone is an amiodarone analogue without iodine. The molecule prolongs atrial and ventricular action potentials and its efficacy is maintained at high heart rates. This drug had deleterious effects when given to patients with coronary artery disease and left ventricular dysfunction. Gastrointestinal side effects may be observed at high dosage. The great advantages of dronedarone are the absence of thyroid complications and of pro-arrhythmic effects. PMID- 15881848 TI - [Angioplasty at the bifurcation of the anterior interventricular artery and diagonal artery]. AB - The IVA/diagonal coronary bifurcation is a high risk area for atheromatous disease. Major technical and strategic risks make the treatment of these lesions a real "challenge" for the interventional cardiologist: angioplasty-stenting and surgery are in direct competition. OBJECTIVES: the aim of this study was to determine the significance of interventional techniques in treating the IVA/diagonal bifurcation, drawing on the experiences of a cardiological haemodynamic laboratory and comparing these results with those obtained with the reference technique, represented by aorto-coronary bypass with the internal mammary artery. METHODS: this was a monocentric retrospective study of a series of 133 patients treated with angioplasty-stenting between January 1997 and December 2002 for a new IVA/Dg bifurcation lesion. Patients admitted for myocardial infarction were excluded. RESULTS: no matter which angioplasty revascularisation technique was used, the angiographic success rate was 95% with 3% occlusions of the diagonal artery. At six months, 72% of patients were asymptomatic, the rate of treated lesion revascularisation (TLR) was 21.9%. At one year 68.8% of patients were asymptomatic, and the TLR was 24.2%. The technique evolved during the six years, especially with the expansion of the radial approach and increasingly frequent use of the "kissing balloon"; at one year the TLR and the rate of major cardiac events was 31% in 1997 and 15% in 2002. CONCLUSIONS: angioplasty-stenting in this at-risk region is thus favourable, and gives results comparable with those of internal mammary/IVA bypass, save on the TLR. However, the development of stents "pre-formed" for the bifurcation and especially "active" endoprostheses could represent a solution to the delicate problem of restenosis. PMID- 15881849 TI - [Echocardiographic indices of diastolic function put to the practical test]. AB - Assessment of left ventricular filling pressures is now part of routine echocardiographic examination. It is a semi-quantitative evaluation usually based on analysis of transmitral blood flow. Restrictive type flow associated with poor left ventricular contraction is pathognomonic of increased left ventricular filling pressure: conversely, inversion of the mitral flow velocities is generally reassuring except in the presence of left ventricular hypertrophy. In other cases, it is necessary to complete the analysis with indices of left ventricular relaxation. The authors discuss the advantages of pulsed Doppler tissue imaging at the mitral ring over the propagation velocity in colour M-mode, especially when left ventricular contraction is normal or relatively preserved. In particular, the reproducibility of propagation velocity measurements is poor when the velocities are relatively high (> 50 cm/s) because a small error in measurement at these velocities has large repercussion on the value of the slope. Pulmonary vein flow, reputedly difficult to analyse, may be very valuable. Discordance of the different indices is one of the most difficult daily problems to resolve. However, it is possible to give a reliable "normal filling pressures" or "increased filling pressures" conclusion in over 80% of cases. Echocardiography cannot provide a precise study of diastole and terms such as abnormal relaxation, abnormal compliance or diastolic dysfunction should be avoided. PMID- 15881850 TI - [Tako-tsubo: a rare form of transient left ventricular dysfunction]. AB - The authors report the case of a 67 year old caucasian female admitted for suspected acute coronary syndrome with ST segment elevation. Coronary angiography did not show any coronary lesion but ventriculography revealed akinesia of the middle and apical segments with hyperkinesia at the base. An isolated moderate elevation of troponine Ic was noted. On the 10th day the methergin test was negative and ventriculography showed complete recovery of the left ventricular segmental kinetics. Left ventricular transient apical ballooning, or tako-tsubo syndrome, is characterised by reversible akinesia of the middle and apical segments with no associated coronary lesion or enzymatic elevation concordant with the extent of the akinetic areas. It mainly affects women between 50 and 60 years old and follows emotional or physical stress. The prognosis is excellent in survivors with a rapid return of normal systolic function. The pathophysiology remains debatable, excluding myocarditis in favour of sideration. PMID- 15881851 TI - [Haemangioma of the right atrium revealed by cardiogenic shock]. AB - Intracardiac haemangioma is a very rare primary benign tumour. A 20 year old female patient, with no significant previous medical history, presented to the emergency department with cardiovascular collapse and vague abdominal pains, with no peripheral signs of cardiac failure. The electrocardiogram showed sinus rhythm with diffuse reploarisation disturbances. Chest radiography revealed cardiomegaly (cardiothoracic index of 0.67) with a right paracardial opacity. Abdominal ultrasound showed a moderate peritoneal effusion and transthoracic ultrasound showed a tumour occupying the right atrial cavity but sparing the interatrial septum. The patient underwent emergency open heart surgery for tumour resection and right atrial wall repair with autologous pericardium. Histology confirmed a haemangioma. Follow-up at one month was uneventful. The clinical, diagnostic and therapeutic features of this case are underlined. PMID- 15881852 TI - [Bilateral carotid aneurysm revealing Takayasu disease]. AB - Extracranial carotid aneurysms caused by Takayasu's arteritis is extremely rare. Their evolutionary risk is dominated by rupture and cerebral ischemia. We report a case of a 23 years old woman presenting bilateral common carotid aneurysm secondary to Takayasu's arteritis. Surgical treatment with the patient under steroid coverage was performed to prevent the risque of rupture. The aneurysm was resected, reconstruction was performed with prothetic aortocarotid bypass. She was discharged without major complications. The graft have remanied patent during the four years follow-up periods. A biologic inflammatory syndrome and a parietal thickening of the aneurysm in a young woman must provoke diagnosis. The surgery must be appropriate every time that the diagnosis is carried considering the evolutionary risk of these aneurysms. PMID- 15881853 TI - Research using animals: an ethical dilemma. PMID- 15881854 TI - Lung functions with spirometry: an Indian perspective--I. Peak expiratory flow rates. AB - Peak expiratory flow rate is an effective measure of effort dependent airflow. It is relatively a simple procedure, and may be carried out in the field using portable instruments. The average PEFR of healthy young Indian males and females is around 500 and 350 lpm respectively. The PEFR reaches a peak at about 18-20 years, maintains this level up to about 30 years in males, and about 40 years in females, and then declines with age. Common regression equations for Indians enveloping major studies from various parts of the country have been formulated. Indian PEFR values compare favourably with other ethnic groups such as Americans and Europeans. PMID- 15881855 TI - Pulmonary blood flow increment and augmentation of ventilation: a review of mechanisms. PMID- 15881856 TI - Effect of (Ala8,13,18)-magainin II amide on human trophoblast cells in vitro. AB - Magainins are cationic peptides with anti-bacterial and anti-tumor properties. The anti-nidatory function of a synthetic analogue of magainin, (Ala8,13,18) magainin II amide, has earlier been reported, and it has been indicated that placental trophoblast cells could be a target of magainin resulting in its contragestational action. The aim of the present study was to examine the effect of (Ala8,13,18)-magainin II amide (100 ng/ml and 1000 ng/ml) on attachment efficiency, viability, differentiation in terms of hCG secretion and invasive function of isolated first trimester, human placental trophoblast cells grown on rat-tail collagen type-I matrix in primary cell culture. In the present experimental model, magainin was not found to affect human trophoblast cell functions in vitro. PMID- 15881857 TI - Effects of dexfenfluramine on dopamine dependent behaviours in rats. AB - 5-hydroxytryptamine (5-HT) inhibits the synthesis and release of dopamine (DA) from rat nigrostriatal DAergic neurons. Dexfenfluramine releases 5-HT from brain 5-HTergic neurons. The present study was undertaken to determine whether dexfenfluramine, through the released 5-HT, modulates the intensity of the behaviours dependent on the functional status of the nigrostriatal DAergic system. The effect of pretreatment with dexfenfluramine on dexamphetamine and apomorphine stereotypies of the oral movement variety and on catalepsy induced by haloperidol and small doses (0.05 and 0.1 mg/kg ip) of apomorphine was studied in rats. We also investigated whether dexfenfluramine induces catalepsy in rats. Dexfenfluramine at 2.5, 5 and 10 mg/kg ip did not induce catalepsy and did not antagonise apomorphine stereotypy. However, 1 h pretreatment with 5-HT releasing doses of dexfenfluramine ie 5 and 10 mg/kg ip, antagonized dexamphetamine stereotypy and potentiated catalepsy induced by haloperidol and small doses of apomorphine. Our results, that dexfenfluramine at 2.5, 5 and 10 mg/kg ip neither induced catalepsy nor antagonised apomorphine stereotypy, indicate that dexfenfluramine at these doses does not block the postsynaptic striatal D2 and D1 DA receptors. They also indicate that the 5-HT released by 5 and 10 mg/kg dexfenfluramine does not exert an inhibitory effect at or beyond the postsynaptic striatal D2 and D1 DA receptor sites. However, 5 and 10 mg/kg doses of dexfenfluramine, through the released 5-HT, inhibit the synthesis and release of DA from the nigrostriatal DAergic neurons and thus antagonise dexamphetamine stereotypy and potentiate catalepsy induced by haloperidol and small doses of apomorphine. PMID- 15881858 TI - Serum lipid response to introducing ghee as a partial replacement for mustard oil in the diet of healthy young Indians. AB - Ghee (clarified butter) has generally been assumed to be hypercholesterolaemic on the basis of its composition but there is hardly any study to support or refute the assumption. The present study was conducted on sixty-three healthy, young, physically active adult volunteers (52 male, 11 female). The study design was that of a randomized controlled trial with a parallel design. After a lead-in period of 2 wk, the subjects were randomly divided into two groups, Group A (n = 30; 25 male, 5 female) and Group B (n = 33; 27 male, 6 female). Group A (experimental) consumed for 8 wk a diet in which ghee provided 10% of the energy intake. The only other visible fat in the diet was mustard oil, and total energy from fats was 25% of the energy intake. Group B (control) consumed for 8 wk a similar diet except that all visible fat came from mustard oil. The serum total cholesterol level showed a significant rise in the experimental group at 4 wk; the rise persisted at 8 wk. A similar rise was also seen in HDL cholesterol. Hence the total cholesterol/HDL cholesterol ratio did not show any significant change. In the control group, there was a trend towards a fall in LDL cholesterol but the change was not significant. The study does not indicate any adverse effect of ghee on lipoprotein profile. However, more studies are needed on older subjects, hyperlipidaemic subjects, and on subjects following less healthy lifestyles before the results of this study can be extrapolated to the general population. PMID- 15881859 TI - Effect of ambroxol, spirulina and vitamin-E in naphthalene induced cataract in female rats. AB - Anticataract activity of Ambroxol, Spirulina and Vitamin E was examined using the naphthalene cataract model. Adult female albino rats of Wistar strain weighing between 180 and 220 grams were taken and divided into eight groups. Group I received light liquid paraffin 5 ml/kg/ day p.o. for 6 weeks. Group II received naphthalene solution 0.5 gm/kg/ day p.o. for first three days and 1 gm/kg/day p.o. thereafter for six weeks. Group III received Ambroxol suspension in 0.5% carboxy methyl cellulose (CMC) at the dose of 100 mg/kg/day p.o. alongwith naphthalene. Group IV received Spirulina in distilled water at the dose of 1500 mg/kg/ day p.o. alongwith naphthalene. Group V received Vitamin E emulsion at the dose of 50 mg/kg/day p.o. alongwith naphthalene. Group VI received Ambroxol alone at the dose of 100 mg/kg/day p.o. Group VII received Spirulina alone at the dose of 1500 mg/kg/day p.o. Group VIII received vitamin E alone at the dose of 50 mg/kg/day p.o. Lens glutathione, soluble protein and water content profiles revealed the preventive role of Ambroxol, Spirulina and Vitamin E in naphthalene induced cataract in female rats. PMID- 15881860 TI - Protective effects of Piper nigrum and Vinca rosea in alloxan induced diabetic rats. AB - In the present study aqueous extract of Piper nigrum seeds and Vinca rosea flowers were administered orally to alloxan induced diabetic rats once a day for 4 weeks. These treatments lead to significant lowering of blood sugar level and reduction in serum lipids. The levels of antioxidant enzymes, catalase and glutathione peroxidase decreased in alloxan induced diabetic rats however these levels returned to normal in insulin, P. nigrum and V. rosea treated rats. There was no significant difference in superoxide dismutase activity in all groups compared to controls. Lipid peroxidation levels were significantly higher in diabetic rats and it was slightly increased in insulin, P. nigrum and V. rosea treated rats as compared to control rat. These results suggest that oxidative stress plays a key role in diabetes, and treatment with P. nigrum and V. rosea are useful in controlling not only the glucose and lipid levels but these components may also be helpful in strengthening the antioxidants potential. PMID- 15881861 TI - Effects of calcium, strontium, and barium on isolated phrenic nerve-diaphragm preparation of rat and their interactions with diltiazem and nifedipine. AB - Calcium (Ca2+), strontium (Sr2+), and barium (Ba2+) are expected to exert similar chemical and pharmacological effects. The effects of barium, strontium and calcium were studied on the contractions of rat phrenic nerve-diaphragm preparations, following electrical stimulation and their interactions with nifedipine (nif) and diltiazem (DZM) were also studied. Low doses of strontium chloride (SrCl2), barium chloride (BaCl2) and calcium chloride (CaCl2) were able to increase the force of contraction of the rat diaphragm when actively stimulated. Diltiazem inhibited the stimulant effects of Sr2+, Ba2+, and Ca2+. On the other hand, nifedipine blocked the effects of Sr2+ and Ca2+ but potentiated the effects of Ba2+. Strontium, barium, and calcium restored the contractility of the muscle following electrical stimulation when the tissue was in biological fluid absolutely depleted of calcium. These findings suggest that Sr2+ and Ba2+ may be able to substitute Ca2+ in the rat diaphragm for its contractions and nifedipine and diltiazem may follow different mechanisms of actions or channels in their blocking effects. PMID- 15881862 TI - CNS alpha 1 receptor in tonic pain during estrous cycle in rats. AB - Estrogen and progesterone are known to affect nociception. The plasma concentrations of these hormones vary during estrous cycle in rodents. The aim of the present study was to investigate the effect of evidence of alpha1 receptor agonist and antagonist on tonic pain in all phases of estrous cycle in female rats. Phenylephrine (alpha1 agonist) and prazosin (alpha1 antagonist) were administered via intracerebroventicular (ICV) injection. Adult female rats weighting 200-220 g were maintained on 12 h light/dark cycle for 10-14 days prior to the experiment. Food and water were made available ad libitum. Formalin test was performed in all phases of estrous cycle. Results showed that phenylephrine caused significant (P<0.05) reduction in pain sensitivity. This reduction was more pronounced during proestrus phase. Prazosin significantly (P<0.05) increased pain sensitivity, particularly during metestrus phase. It is possible that fluctuation in pain sensitivity during estrous cycle is related to the level of sex hormones during estrous cycle. PMID- 15881863 TI - Management of chronic hepatitis B with New Livfit in end stage renal disease. AB - New Livfit (NLF) is a standardized, poly-herbal formulation that has been found useful in the management of hepatitis. The aim of this placebo-controlled study was to evaluate its usefulness against hepatitis B virus in the patients of end stage renal disease (ESRD). Patients were regularly evaluated at 6, 12, 24 and 36 weeks of therapy. With 36 weeks of treatment of NLF, there was rapid clearing of HBV-DNA in a significant number of patients. Significant seroconversion of the other markers of hepatitis B and restoration of the raised levels of ALT and AST was observed. The study suggests the potential usefulness of NLF in the control of HBV infection in the patients of ESRD prior to renal transplant. PMID- 15881864 TI - Sympathetic hyperactivity in patients of bronchial asthma. AB - The bronchoconstricting and bronchodilating divisions of the autonomic nervous system control airway caliber of lungs. Parasympathetic hyperactivity in asthmatics eventuates in broncho-constriction. Sympathetic activity increases to combat this but fails to control bronchoconstriction due to negligible innervations of the airway smooth muscles although it causes vasoconstriction of blood vessels. In this study the effects of autonomic function test for sympathetic division were seen on selected patients and controls. Our results indicate sympathetic hyperactivity or an increased alpha-adrenergic input in asthmatics. PMID- 15881865 TI - Oxidative stress and protein glycation in patients with chronic obstructive pulmonary disease. AB - Several studies have indicated the presence of increased oxidative stress as a critical feature in the pathogenesis of chronic obstructive pulmonary disease (COPD). Another biochemical complication leading to pathogenesis is protein glycation. The nexus between oxidative stress and protein glycation in various pathological conditions is being unraveled. Increased oxidative stress can lead to enhanced protein glycation by a process of auto-oxidative glycation. No information is available in the literature regarding protein glycation among COPD patients. Eleven non-diabetic COPD patients were included in the study and equal number of age and sex-matched healthy individuals were enrolled as controls. The whole-blood reduced glutathione was found to be less among the patients while lipid peroxides and fructosamine were elevated in comparison to control. The present study confirmed oxidative stress and enhanced protein glycation among the COPD patients. Antioxidant therapy may be considered as part of the treatment regimen for COPD patients. PMID- 15881866 TI - Salt preference across different phases of menstrual cycle. AB - It is well established that women experience food craving for particular foods and gain weight in relation to phases of menstrual cycle. In this study, the preference for different concentrations of salt sprayed on bland popcorn was assessed in 55 healthy women (age 18 to 22 yrs). Salt solutions of 0, 1, 2, 3 and +3 molar strength were used. Samples of sprayed popcorn were consumed in random order and preference marked on a Likert scale. It was observed that women preferred unsalted popcorn in the menstrual phase more than in the luteal phase. The preference for salted popcorn was most during the luteal phase and was proportionate to the strength of the salt solution used. Statistical analysis revealed significant differences in the preference rating between the menstrual phase and the other two phases. There was no significant difference in preference between the luteal and follicular phases. PMID- 15881867 TI - Atenolol or butoxamine injection at the lateral septum doesn't inhibit male sexual behavior in rats. AB - To investigate the role of specific adrenoreceptors subtypes on sexual behavior, atenolol, butoxamine, a mixture of atenolol and butoxamine, and saline (vehicle) were injected into the lateral septum in four different groups of sexually active male rats. Application of a mixture of atenolol and butoxamine produced inhibition of copulatory activity. On the other hand, application of either atenolol or butoxamine alone did not inhibit copulatory activity. But it produced stimulation of some of the components of male sexual behavior. Inability of either atenolol or butoxamine to inhibit the male sexual behavior, and inhibition of the same by the mixture of atenolol and butoxamine, indicate that both beta adrenoreceptors at the lateral septum are involved in the elaboration of male sexual behavior. Stimulation of some components of sexual behavior on application of atenolol or butoxamine could be attributed to an unbalanced activity of beta adrenoreceptors. PMID- 15881868 TI - Cerebrospinal fluid lactate dehydrogenase and glutamine in meningitis. AB - The cerebrospinal fluid concentration of Glutamine and Lactate dehydrogenase (LDH) were studied in patients with pyogenic and tubercular meningitis. Significant increase in Glutamine and LDH level (P<0.001) were observed in the test group when compared to the control group. LDH and glutamine may not be useful in differentiating viral from other meningitis. It may act as corroborative evidence of meningitis. PMID- 15881869 TI - Protective effect of curcumin during selenium induced toxicity on dehydrogenases in hepatic tissue. AB - Selenium administration resulted in a marked decrease in the activity levels of the liver succinate dehydrogenase, malate dehydrogenase, and lactate dehydrogenase while pyruvate dehydrogenase increased significantly (P<0.001) in the wistar rat. The degree of decrease of these enzymes was significantly less (P<0.001) when rats were treated with curcumin, a natural constituent Curcuma longa. Curcumin seems to prevent oxidative damage mediated through selenium and protect the dehydrogenases possibly through its anti-oxidative property. PMID- 15881870 TI - Effect of garlic (Allium sativum) oil on exercise tolerance in patients with coronary artery disease. AB - The effect of six weeks garlic oil administration was observed on cardiac performance and exercise tolerance in 30 patients of coronary artery disease. After initial treadmill stress test, they were administered garlic oil in the dose of four capsules twice a day for 6 weeks and treadmill stress test was repeated. Garlic significantly (P<0.01) reduced heart rate at peak exercise and also significantly reduced the work load upon the heart resulting in better exercise tolerance (P<0.05) as compared to the initial test. It appears to be a good adaptogen to be utilized in patients with coronary artery disease. PMID- 15881871 TI - Identification of susceptibility to hypertension by the cold pressor test. PMID- 15881872 TI - Nobel Prize in Chemistry, 2004. Aaron Ciechanover, Avram Hershko and Irwin Rose. PMID- 15881873 TI - Nobel Prize in Physiology or Medicine, 2004. Richard Axel and Linda B Buck. PMID- 15881874 TI - [Role of the human thalamic parafascicular (CM-Pf) complex in neuronal mechanisms of selective attention]. AB - Responses of 93 single units of the human thalamic CM-Pf complex to relevant and irrelevant verbal (or sensory) stimuli were studied using microelectrode technique in alert diskinetic patients suffering from the tonic forms of spasmodic torticollis during 11 stereotaxic operations. The response patterns of two types units with irregular unitary (A-type) and low-threshold bursting Ca(2+) dependent (B-type) spike activity were studied. Three main conclusions emerge from the studies: 1) high reactivity of both A- and B-units to presentation of relevant verbal stimuli with differences of their response patterns as determined by the type of constituent elements; 2) close functional connectivity of these neuronal changes with the level of selective attention; 3) at the moment of attention activation, the appearance of transient interneuronal interactions between adjacent A and B cells characterized by the local synchronization and stabilization of rhythmic oscillations. These data point to considerable contribution of the thalamic CM-Pf complex and its neuronal mechanisms into organization of the human selective attention and triggering verbal-related processing during performance of purposive speech-provoked voluntary acts. PMID- 15881875 TI - [The effect of ethanol consumption by dams on the offspring locomotion in the open field test and carboxypeptidase activities in the rat brain and adrenal medulla]. AB - Consumption of dams ethanol increased the posterity locomotion activity in open field test. The increase in female rats was higher then in male ones. Differences in the carboxypeptidase H and PMSF-inhibited carboxypeptidase activities between the brain regions and adrenal medulla of prenatally exposed to ethanol and intact rats were found. The changing of enzyme activities in female rats was higher then in male ones. It is possible that dams ethanol consumption induced profound changes in locomotion mediated, at least partially, by changes in the rate of proteolytic processing of neuropeptide precursors. PMID- 15881876 TI - [Vestibular effects on posture instability evoked by moving visual environment and footing]. AB - Subjects held the vertical posture standing up on hard footing, having small degree of the freedom in the frontal plane. The stability of the vertical posture has been assessed by the standard deviations (sigma) from average amplitudes of the fluctuations of the subject's head (in frontal and sagittal planes) from conditional zero. Sinusoidal rotations of optokinetic cylinder, sinusoidal rotations of the footing, and combinations of these rotations, under phase shifts between the optokinetic cylinder and the footing, caused increase of sigma. The amplitude and velocity signal of the head deviations was transformed into low galvanic current applied to the mastoids and used as the artifical vestibular biofeedback. It was possible to reduce the value of the sigma for lateral tilts (raised in comparison with their values during stance in the dark as a result of destabilizing influence), varying coefficients of the biofeedback. At the same time, appropriate fluctuations in sagittal plane were not systematic. PMID- 15881877 TI - [Changes in hemodynamics of the long bone due to tunnelization under pulsing and non-pulsing pressure in a shaft cavity]. AB - In 25 dogs, it was revealed that the pressure in a long bone shaft cavity (IBP) pulsed at relatively high values. According to rheovasography data, the pulse changes of the vascular blood filling of a shaft cavity caused the IBP. The blood filling changes at pulsing IBP produced pulse changes in interstitial fluid amount, at non-pulsing IBP they provided for fluid coming into the interstitial space of the shaft cavity without pulsations. Tunnelization led to appearance of compact substance defect, at the size of which, coming to 2.4% of shaft area, IBP always decreased, remained decreased during 2 months and restored during the third month. The IBP decrease changed the bone hemodynamics. PMID- 15881878 TI - [Presynaptic effects of arachidonic acid and prostaglandin E2 in the frog neuromuscular synapse]. AB - Arachidonic acid and prostaglandin E2 decreased the frequency of miniature endplate currents without changing their amplitude-temporary parameters. They also reduced the evoked transmitter release and the amplitude of the 3rd phase of nerve ending response corresponding to the voltage-dependent K(+)-current. Using perineural recording, It was shown that arachidonic acid and prostaglandin E2 decreased the Ca2+ currents of nerve endings. Indometacin: inhibitor of cyclooxygenase, enhanced the evoked transmitter release and decreased the 3rd phase of nerve ending response. Indometacin prevented the effects of arachidonic acid on evoked transmitter release, whereas the effects of arachidonic acid on the 3rd phase was preserved. Prostaglandin E2 seems to mediate the effects of arachidonic acid on spontaneous and evoked transmitter release, Ca(2+)- and Ca(2+)-activated K(+)-currents. Moreover, the arachidonic acid and prostaglandin E2 exerted their own effects upon voltage-dependent potassium current of motor nerve ending. PMID- 15881879 TI - [Is stress-induced analgesia a measure of stress severity?]. AB - When comparing magnitudes of "behavioural despair" (duration of immobility) and stress-induced analgesia in the tail suspension test and cold water swim test with SHR and NMRI male mice. The results might depend on saline injection prior the test and on the fact that exposure to cold water in swim test was sufficient to alter the response patterns. The findings show that the main parameters are closely related to each other. Stress-induced analgesia seems to be a measure of stress as the stress becomes stronger analgesia changes in linear dependence, whereas duration of immobility has an invert U-shaped function. PMID- 15881880 TI - [Stimulation of physiological functions in cooled rats without rewarming after introducing ethylenediaminetetraacetate into lateral ventricles of the brain]. AB - On cooling the animals to the rectum temperature from approximately 37 to 24-28 degrees C the decreases were shown to occur in the frequency and amplitude of respiration motions, in the intensity of muscle electrical activity (thermoregulation muscle tone and cold muscle shivering), in the frequency of heart contractions. In 3-8 min after introducing ethylenediaminetetraacetate into a lateral ventricle of the rat brain the frequency and the amplitude of respiration motions increased statistically reliable and so did the intensity of thermoregulation muscle tone and cold muscle shivering (judging from the total muscle electrical activity). The doses of EDTA, which caused this effect, were by a factor of 30 - 100 less than the doses, which caused a similar stimulation of the functions in cooled animals after introduction into the blood. PMID- 15881881 TI - [Rectal temperature in active and passive rats during desynchronosis and under melatonin treatment ]. AB - Effects of phase shifts in circadian rhythms and of melatonin administration on rectal temperature in rats with different activity were studied in the open-field test on 176 Wistar rats kept under conditions of natural or shifted light darkness period. Under normal light-darkness conditions, the amplitude of diurnal variation in rectal temperature was higher in active rats as compared with passive ones. A shift in the light-darkness conditions inverted the circadian rhythm of rectal temperature and augmented the difference between daytime and night time temperatures in passive and, particularly, in active rats. Melatonin effect depended on dose and time of administration. 1 mg/kg Melatonin enhanced the amplitude of diurnal rhythms of energy metabolism in behaviourally active rats. These changes seem to contribute to adaptive reconstruction in the organism during desynchronosis. PMID- 15881882 TI - [The GABA-ergic regulation of the Lymnaea stagnalis (L.) dorsal longitudinal muscle contractions]. AB - Presence of GABA-immunoreactive fibres in the nerve cervicalis inferior was shown as well as their regulatory effects upon contractions of the dorsal longitudinal muscle in Lymnaea stagnalis. The exogenous GABA affected the amplitude of evoked muscle contractions diminishing them at concentrations from 10(-8) M and augmenting them from above 10(-5) M. Picrotoxin produced opposite effects on the muscle contractions in the same concentration range. Bicucullin did not affect the contractions. The findings are discussed from the standpoint of the supposed functional heterogeneity of the GABA receptors in the mollusc nerve-muscle junction. PMID- 15881883 TI - [Defensin NP-1 in restoration of the injured nerve trunk functions]. AB - The effect of Defensin NP-1 on early phase of injured sciatic nerve regeneration was studied in rats. Using the technique of AP compound recording performed within 21 days after the nerve trunk cutting with subsequent microsurgical nerve suture, the rate of fibres growth was shown to increase by 30% following Defensin treatment: the distance to which the nerve impulse conductivity through injured nerve fibres was restored has extended from 7.22 +/- 1.2 mm (control) to 10.5 +/- 0.8 mm (Defensin treatment) from the suture site (p < 0.05). Moreover, the conducting capacity of the regenerating nerve fibres has risen by 20% as compared with control values. The findings suggest a positive effect of Defensin on restoration of injured peripheral nerve in rats. PMID- 15881884 TI - [Effect of estragole on glucocorticoid induction of tyrosine aminotransferase and tryprophan oxygenase in the rat and mouse liver]. AB - A single intraperitoneal injection of Estragole (300 mg/kg) to female ICR mice 19 hours prior to Dexamethasone induction decreased induced activities of tyrosine aminotransferase (TAT) and tryptophan oxygenase (TO) nearly to 50% of the control values. In these mice, activities of the marker enzymes of liver damage: alanine aminotransferase (ALAT) and aspartate aminotransferase (AAT) increased in the blood 1.7-2.3-fold as compared with the untreated controls. By contrast, carbon tetrachloride (100 mg/kg) increased the blood AIAT and AsAT activities 135- and 30-fold as compared with the control, but inhibited the TAT and TO induction much less than Estragole did. Estragole seems to inhibit the glucocorticoid induction of these hepatic enzymes not via the unspecific toxic damage of the liver. PMID- 15881885 TI - [Pacemaker activity in distal parts of the cat ureter]. AB - Existence of two types of uncoordinated spontaneous activities of the cat ureterbladder area was shown: spikes and slow waves. The oscillation frequency of this area is two-fold lesser than the rhythm of renal waves. The activity concordance of these pacemakers occurs under conditions of ureter dissection in the middle. Normally pacemakers of this area seem to be a reserve mechanism of urine pushing through. PMID- 15881886 TI - [Concentration of hydrochloric acid and pepsin in gastric juice in dogs after starvation and refeeding]. AB - Feeding fogs with meat after a 3-day period of starvation increased hydrochloric acid concentration with subsequent return of the parameter to normal values. Under the same conditions, pepsin concentration decreased and raised up after re feeding. Histamine administration following the starvation decreased hydrochloric acid concentration with subsequent normalising. In three days after re-feeding and histamine administration, pepsin concentration drooped owing, probably, to a decrease of parietal cell H2-receptor affinity to histamine. Pentagastrin administration after the starvation increased hydrochloric acid concentration. The findings suggest G-cell function inhibition occurring after a 3-day starvation which is important for the stomach mucous membrane protection. PMID- 15881887 TI - [The effect of glucose on antidiuretic hormone absorption in the rat and frog small intestine]. AB - Administration of 5 ml/100 g body weight of 1% glucose solution to stomach produced the same diuretic kidney response in fasted Wistar rats as administration of the same amount of water. Intragastric administration of arginine vasopressin along with the water load evoked an antidiuretic response. Arginine vasopressin in the same volume of glucose induced no kidney response difference as compared with the hormone action in experiments with water load. 0.1 nmol of arginine vasotocin, having been itroduced into the rat isolated ileum, prevented the effect of glucose on the hormone absoption. 0.1 nmol of arginine vasotocin, having been introduced into the frog isolated ileum along with isotonic glucose solution, increased the hormone absorption; fructose did not affect this process whereas mannitol decreased absorption ofarginine vasotocin. This absorption was also reduced by intraileal introduction of arginine vasotocin with the hypotonic Ringer solution. The findings suggest that glucose in the rat gastrointestinal tract does not affect arginine vasopressin absorption in vivo, whereas in the frog ileum glucose increases arginine vasotocin absorption in vitro. PMID- 15881888 TI - [Three-dimensional electrical fields around the snail body during activation of giant neurons in suboesophageal ganglia of Achatina fulica]. PMID- 15881889 TI - Chromatin remodeling factors and DNA replication. AB - Chromatin structures have to be precisely duplicated during DNA replication to maintain tissue-specific gene expression patterns and specialized domains, such as the centromeres. Chromatin remodeling factors are key components involved in this process and include histone chaperones, histone modifying enzymes and ATP dependent chromatin remodeling complexes. Several of these factors interact directly with components of the replication machinery. Histone variants are also important to mark specific chromatin domains. Because chromatin remodeling factors render chromatin dynamic, they may also be involved in facilitating the DNA replication process through condensed chromatin domains. PMID- 15881890 TI - Epigenetic inheritance of chromatin states mediated by Polycomb and trithorax group proteins in Drosophila. AB - Proteins of the Polycomb group (PcG) and of the trithorax group (trxG) are involved in the regulation of key developmental genes, such as homeotic genes. PcG proteins maintain silent states of gene expression, while the trxG of genes counteracts silencing with a chromatin opening function. These factors form multimeric complexes that act on their target chromatin by regulating post translational modifications of histones as well as ATP-dependent remodelling of nucleosome positions. In Drosophila, PcG and trxG complexes are recruited to specific DNA elements named as PcG and trxG response elements (PREs and TREs, respectively). Once recruited, these complexes seem to be able to establish silent or open chromatin states that can be inherited through multiple cell divisions even after decay of the primary silencing or activating signal. In recent years, many components of both groups of factors have been characterized, and the molecular mechanisms underlying their recruitment as well as their mechanism of action on their target genes have been partly elucidated. This chapter summarizes our current knowledge on these aspects and outlines crucial open questions in the field. PMID- 15881891 TI - How to pack the genome for a safe trip. AB - The transformation of the somatic chromatin into a unique and highly compact structure occurring during the post-meiotic phase of spermatogenesis is one of the most dramatic known processes of chromatin remodeling. Paradoxically, no information is available on the mechanisms controlling this specific reorganization of the haploid cell genome. The only existing hints suggest a role for histone variants, as well as for stage-specific post-translational histone modifications,before and during the incorporation of testis-specific basic nuclear proteins. Moreover, the exact functions of the latter remain obscure. This chapter summarizes the major chromatin-associated events taking place during the post-meiotic differentiation of male haploid cells in mammals and discusses some of the basic issues that remain to be solved to finally understand chromatin remodeling during spermatogenesis. PMID- 15881892 TI - Chromatin modifications on the inactive X chromosome. AB - In female mammals, one X chromosome is transcriptionally silenced to achieve dosage compensation between XX females and XY males. This process, known as X inactivation, occurs early in development, such that one X chromosome is silenced in every cell. Once X-inactivation has occurred, the inactive X chromosome is marked by a unique set of epigenetic features that distinguishes it from the active X chromosome and autosomes. These modifications appear sequentially during the transition from a transcriptionally active to an inactive state and, once established, act redundantly to maintain transcriptional silencing. In this review, we survey the unique epigenetic features that characterize the inactive X chromosome, describe the mechanisms by which these marks are established and maintained, and discuss how each contributes to silencing the inactive X chromosome. PMID- 15881893 TI - Chromatin mechanisms in Drosophila dosage compensation. AB - Dosage compensation ensures that males and females equalize the expression of the X-linked genes and therefore provides an exquisite model system to study chromosome-wide transcription regulation. In Drosophila, this is achieved by hyper-transcription of the genes on the male X chromosome. This process requires an RNA/protein-containing dosage compensation complex. Here, we discuss the current status of the known Drosophila complex members as well as the recent views on targeting, assembly and spreading mechanisms. PMID- 15881894 TI - DNA methylation in epigenetic control of gene expression. AB - Over three decades ago DNA methylation had been suggested to play a role in the regulation of gene expression. This chapter reviews the development of this field of research over the last three decades, from the time when this idea was proposed up until now when the molecular mechanisms involved in the effect of DNA methylation on gene expression are becoming common knowledge. The dynamic changes that the DNA methylation pattern undergoes during gametogenesis and embryo development have now been revealed. The three-way connection between DNA methylation, chromatin structure and gene expression has been recently clarified and the interrelationships between DNA methylation and histone modification are currently under investigation. DNA methylation is implicated in developmental processes such as X-chromosome inactivation, genomic imprinting and disease, including tumor development. This chapter discusses all these issues in depth. PMID- 15881895 TI - The epigenetic breakdown of cancer cells: from DNA methylation to histone modifications. AB - The recognition of epigenetic defects in all types of cancer has represented a revolutionary achievement in cancer research in recent years. DNA methylation aberrant changes (global hypomethylation and CpG island hypermethylation) were among the first events to be recognized. The overall scenario comprises a network of factors in which deregulation of DNA methyltransferases leads to a cancer-type specific profile of tumor suppressor genes that become epigenetically silenced. Over recent years, a better understanding of the machinery that connects DNA methylation, chromatin and transcriptional activity, in which histone modifications stand in a key position, has been achieved. The identification of these connections has contributed not only to understanding how epigenetic deregulation occurs in cancer but also to developing novel therapies that can reverse epigenetic defects in cancer cells. PMID- 15881896 TI - Developmental regulation of the beta-globin gene locus. AB - The beta-globin genes have become a classical model for studying regulation of gene expression. Wide-ranging studies have revealed multiple levels of epigenetic regulation that coordinately ensure a highly specialised, tissue- and stage specific gene transcription pattern. Key players include cis-acting elements involved in establishing and maintaining specific chromatin conformations and histone modification patterns, elements engaged in the transcription process through long-range regulatory interactions, transacting general and tissue specific factors. On a larger scale, molecular events occurring at the locus level take place in the context of a highly dynamic nucleus as part of the cellular epigenetic programme. PMID- 15881897 TI - Epigenetic regulation of mammalian imprinted genes: from primary to functional imprints. AB - Parental genomic imprinting was discovered in mammals some 20 years ago. This phenomenon, crucial for normal development, rapidly became a key to understanding epigenetic regulation of mammalian gene expression. In this chapter we present a general overview of the field and describe in detail the 'imprinting cycle'. We provide selected examples that recapitulate our current knowledge of epigenetic regulation at imprinted loci. These epigenetic mechanisms lead to the stable repression of imprinted genes on one parental allele by interfering with 'formatting' for gene expression that usually occurs on expressed alleles. From this perspective, genomic imprinting remarkably illustrates the complexity of the epigenetic mechanisms involved in the control of gene expression in mammals. PMID- 15881898 TI - Seed development and genomic imprinting in plants. AB - Genomic imprinting refers to an epigenetic phenomenon where the activity of an allele depends on its parental origin. Imprinting at individual genes has only been described in mammals and seed plants. We will discuss the role imprinted genes play in seed development and compare the situation in plants with that in mammals. Interestingly, many imprinted genes appear to control cell proliferation and growth in both groups of organisms although imprinting in plants may also be involved in the cellular differentiation of the two pairs of gametes involved in double fertilization. DNA methylation plays some role in the control of parent-of origin-specific expression in both mammals and plants. Thus, although imprinting evolved independently in mammals and plants, there are striking similarities at the phenotypic and possibly also mechanistic level. PMID- 15881899 TI - Nursing's verdict could win or lose the election for Labour. PMID- 15881900 TI - The reality of nursing behind the headlines. PMID- 15881901 TI - Nurses' verdict on Labour. PMID- 15881902 TI - In short supply. Independent nurses lack equipment. PMID- 15881904 TI - What you need to know about coeliac disease. PMID- 15881903 TI - The Children Act: key points and implications for nursing. AB - This article sets out the key points of the Children Act 2004 and considers the implications for practice, with particular regard to the patient assessment process and the links between adult behaviour and child protection. PMID- 15881905 TI - How to manage diabetes in advanced terminal illnesses. AB - Diabetes mellitus is increasing in incidence and is therefore being encountered more frequently during palliative care. Diabetes management in the early stages of terminal disease is normally the same as conventional care. However, in the later stages of terminal illness there are a number of problems that may arise due to diabetes treatment, including hypoglycaemia and hyperglycaemia, which may impair the patient's quality of life. This article will address some of these problems and their management. PMID- 15881906 TI - Drugs. Adapalene. PMID- 15881907 TI - Effectiveness of a nurse-led pregnancy termination clinic. AB - A new nurse-led service was developed for women requesting pregnancy termination. Compared with the previous service, the nurse-led clinic reduced waiting times and cost almost 40 per cent less. PMID- 15881908 TI - Mobility and safe handling of people with dementia. AB - Moving and handling patients can be a serious risk to health care staff, and requires careful risk assessment to minimise the likelihood of harm. People with dementia can be particularly challenging because immobility can have a number of causes other than simple lack of cooperation. PMID- 15881909 TI - Improving diagnosis and treatment for appendicitis. PMID- 15881910 TI - Leading the school nursing revolution. School nursing is a prime example of how nurses' roles in the community are changing. PMID- 15881911 TI - [Ancient urology in Galicia (Spain)]. AB - INTRODUCTION: Although Urology is a recent branch of medical science and practice, it is not a contradiction to talk about ancient Urology, as a reflection on the ways of getting sick and treating genitourinary diseases in past times, sufferings that are recognized if compared to the present-day ones. This paper takes into account the social view of the history of medicine and tries to reject either the intention to create a mythic past for urology or to underestimate the ancient practices because of irrational or not scientific. MATERIAL Y METHOD: Review based on primary and secondary sources, with reference to the medicine and surgery of the urinary tract and genitalia in Galicia (Spain) and the Iberian peninsula, from the ancient and medieval periods, to the appearance of modern, anatomo-clinic medicine in the XVIIIth century. RESULTS: Because of the pilgrimage through the Way of Saint James to Santiago de Compostela, many centers of great importance for the attention of sick people rose up in Galicia, like the Monastery of Saint Martin Pinario and the Hospital of the Catholic Kings, where traditional European standars of patient care and use of medicinal plants were followed in. Ancient medicine and surgery of the urinary tract and genitalia had in Galicia, characters of great interest, such as doctor Romano, instructor of Galician practitioners in catheterisation and urethral dilatation, and important thinkers such as the physician- philosopher Francisco Sanchez and the erudite monks Sarmiento and Feijoo, whom, without being doctors, may be considered as pioneers of modern evidenced-based medicine. COMMENTS: The importance that the Galician hospitals had in past times and the works of several characters related to Galician country, had a distinguished place in the ancient history of the medicine and surgery of the urinary tract and genitalia. PMID- 15881912 TI - [Scientific evidence on the use of high-intensity focal ultrasound (HIFU) in the treatment of prostatic carcinoma]. AB - OBJECTIVES: To evaluate in the literature scientific evidence on the use of High Intensity Focal Ultrasound (HIFU) in the treatment of prostatic carcinoma (PC). METHOD: Three database are searched: PubMed, Cochrane Library, HTA database. Several articles were selected taking into account number of cases, inclusion criteria, duration of follow-up period. We have evaluated the best evidence available through a systematic review of clinical efficacy and cost-effectiveness of HIFU in the treatment of PC. We analized global survival, disease-free survival, and quality of life, including complications, adversal effects and acceptance of the technique. RESULTS: Publications available are focused on two main indications of the therapy: first step of management of PC and salvage therapy for locally recurrent PC after external beam radiotherapy. It was very difficult to draw conclusions on the relative benefits of the HIFU: lack of high or medium quality evidence and no comparisons between this technique an standard treatments. In relation to results on cost-effectiveness, no relevant studies were identified in order to get conclusions on the quality of the treatment. Most of reports offered disparity in the definition of free survival disease concept. This fact produce some misunderstanding of results and conclusions cannot be drawn correctly. Inclusion criteria were also heterogeneous between authors. CONCLUSIONS: No high-quality clinical evidence can be established currently on the utility of HIFU as treatment of prostatic cancer. An important fact to stress is the capacity of therapy to produce tumour necrosis both as first-step treatment and as salvage therapy. No conclusions can be drawn in the long-term due to the paucity of controlled and randomized trials with adequate follow-up to establish benefits in terms of global survival and quality of life (balance adversal effects/benefits), lack of comparisons with standard options as long as different definitions of free-survival disease. PMID- 15881913 TI - [Primary vesicoureteral reflux treatment in childhood: comparsion of two systematic review]. AB - Many medical practices are being carried out unawares of their efficiency, or of their actual impact on the health of the patients, therefore it is necessary to consider the support of professional recommendations with scientific evidence. THE PRIMARY OBJECTIVE: To perform a systematic review (SR) of the therapeutic management of primary VUR in pediatric urology. MATERIAL AND METHODS: A systematic review has been performed, including scientific evidence-based medicine criteria, of the articles published in all of the available databases. Inclusion criteria concerning basic quality of the articles were considered essential, as well as exclusion criteria to be able to reject the articles. RESULTS AND DISCUSSION: Subsequently, and following the critic reading of greater than 320 articles, statistical study of the grouped data was performed according to the type of treatment and to the benefits contributed by each treatment, and also to their undesirable effects. Finally we have made a comparison between our results and recent Cochrane Systematic Review. The following Conclusions were drawn from the results obtained and from the analysis of the texts. Both medical and surgical treatment present with similar effectiveness concerning resolution of grades I, II and III of VUR, and the former one is the recommended initial treatment following diagnosis. Endoscopic treatment is exactly as effective as open surgery for grades I, II and III with fewer undesirable effects secondary. There are no differences concerning the efficacy of the different injected substances. Not enough evidences exist for degrees IV and V that may recommend or advise against any of the treatments. In any degree of VUR, open surgical treatment is superior as far as medical treatment is concerned only regarding the number of acute pyelonephritis episodes during followup. This conclusion cannot be applied on endoscopic treatment. PMID- 15881914 TI - [Porphyrins in renal calculi (review)]. AB - We performed a review of the porphyrines content in a type of black, charcoal like, renal calculi exhibiting infrared spectra (IRS) similar to those characteristic of "organic material" which has not yet been fully elucidate. Several other types of renal calculi, mainly those of small size, spontaneous passage renal stone, may also have diffuse or isolated dark charcoal components showing "organic material" IRS. After observing that haemoglobin has an "organic material" IRS, we studied, by a sensitive thin layer chromatography method, the presence of porphyrines in several types of dark or charcoal renal stones, since porphyrines are the physiological precursors of haemo group biosynthesis. We found two types of porphyrine content: coproporphyrin, in patients suffering from hepatopathy, and uroporphyrin and heptacarboxil-porphyrin in patients with some types of porphyria or with chronic renal failure. PMID- 15881915 TI - [Role of chronic prostatic infiltrate from specimens of prostate biopsy in total and free PSA determination]. AB - PURPOSE: To analyze the influence in total serum PSA (PSA(t)) and free PSA (PSA(1)) of chronic inflammatory patterns from prostate biopsy specimens of non symptomatic patients. PATIENTS AND METHODS: 518 non-symptomatic patients underwent ultrasonography and prostatic biopsy for PSA(t) > 4 ng/mL and/or DRE suspicious of malignancy. Those with a negative biopsy were divided into two subgroups: patients with benign prostatic lesions (LBP) and patients with chronic inflammatory signs (SIC). RESULTS: 456 patients (88.03%) were biopsied for elevated PSA(t) or/and DRE suspicious of malignancy in 62 patients (11.97%). Mean volume in patients with LBP was 54cc, while 51cc was the mean volume in patients with histological pattern of chronic inflammatory infiltrate. Mean PSA(t) in patients with LBP was 9.43 (IC 95% +/- 7.8) and 8.8 (IC 95% +/- 5.73) in SIC's group patients. CONCLUSIONS: The presence of SIC has no significant influence in the eventual value of PSA(1) y PSA(t) in a selected population. As a result of that finding data from serum PSA can be reliably evaluated even in the presence of SIC in biopsy specimens. PMID- 15881916 TI - [Analysis of the absorcion's factors of irrigating fluid during transurethral resection of the prostate]. AB - OBJECTIVES: In this study it was our intention to evaluate the relation between the reabsorption of irrigating fluid and three variables: time of the intervention, volume of solution of glicina employee and weight of the fragments, during the RTU of prostate fulfilled to low hydraulic pressure. MATERIAL AND METHOD: We study 74 patients that RTU of prostate was performed with suprapubica derivation with Amplatz's pod 30 ch. The ethanol was monitored in expired air every 15 minutes during the intervention. Likewise we annotated the time of the intervention, the volume of glicina used and the weight of the fragments extracted. Statistically Anova's text was in use for comparison of averages. RESULTS: 13,6% of the patients absorbed irrigating fluid in some quantity. The range of absorption belongs to 100 cc until 2.000 cc. We did not find a statistically significant difference in the averages of time of resection, volume of glicina and weight of the fragments between the group of patients that had absortion of irrigating fluid and they that didn't had. CONCLUSIONS: Our data show that the operative time, the volume of irrigating fluid and the weight of the resected fragments do not influence the reabsorption of liquid of irrigation when a RTU is realized to low hydraulic pressure. PMID- 15881917 TI - [Survival prognostic factors valuation on a series of 202 patients with surgical treatment of renal cell carcinoma]. AB - OBJECTIVES: To describe renal cell carcinoma prognostic factors and set up the relationship with survival rates in this neoplasm. Likewise we show epidemiologic, clinical, diagnosis and therapeutic facts. MATERIAL AND METHOD: We review 202 patients underwent surgical treatment for renal cell carcinoma and the following features were recorded: gender, age and presenting symptoms, especially incidentally discovered tumors; tumor-related factors like TNM tumor stage, tumor grade and venous involvement: therapy-related recorded were surgical techniques and cytokine-based therapy. RESULTS: 60% of the patients showed organ-confined disease, 10% of patients with renal cell carcinoma presented with nodal positive disease and 7% with systemic metastases. 42% of patients presenting incidental tumor, with survival rates substantially better than that for symptomatic patients. 42% of patients with nodal positive disease presented systemic metastases at diagnosis, and 30% at surveillance. Systemic metastases presented a particularly poor prognosis for patients with renal cell carcinoma, with 12 months survival rates that 0%. Patients with cytokine-related therapy for metastatic disease presented 24-months survival rates that 20%. CONCLUSIONS: Renal cell carcinoma remains a major source of mortality, basically at advanced disease (nodal positive disease or systemic disease), without a clear improvement of survival rates despite the newer therapy modalities. PMID- 15881918 TI - [Role of nephrectomy in metastatic renal cell carcinoma. Experience of the Department of Urology Juan Canalejo Hospital]. AB - OBJECTIVES: We expose our experience in nephrectomy in metastatic renal cell carcinoma, and also show complications, evolution and survival of these patients. MATERIAL AND METHODS: We performe a retrospective review of renal cell carcinoma treated at our service in the period between January 1st 1991 and December 31st 2002. We only studied those which presented in a metastatic pattern (31), and divide these in two groups: the ones which were nephrectomized and those which were not. We try to showw the differences between the two groups in order of status performance (E.C.O.G.), associated morbidity and median survival. In the first group we also study complications of surgery and treatment that patients underwent. RESULTS: we performed nephrectomy in 19 cases, all of them E.C.O.G. 0 1. Median postoperative stay was 12 days, and complication rate was 11.5%. Of these patients, 45% underwent some type of systemic treatment, and median survival was 31 months. We didn't performed nephrectomy in 12 patients, of which 9 were E.C.O.G. 2-3. Associated co-morbidity was higher in this group. Only in three patients any treatment was offered always with palliative reason. Median survival was 3.8 months. CONCLUSIONS: In those patients with good performance status this approach does not represent more morbility nor mortality than in non metastatic patients, and that is a cornerstone in their management. We also make a literature review in which we see the last pathways in the management of these patients, and that show the needing for a combined approach both quirurgical and inmunotherapical. We have review with special interest the studie's conclusions of SWOG and EORTC groups. PMID- 15881919 TI - [Immunohistochemical and molecular analyis of mismatch repair genes in germ cell tumors]. AB - Correction of misincorporated nucleotides during DNA replication (mismatch repair) distinguishes histologically similar cancers with distinct biological and clinical behavior. We investigated expression of two mismatch repair genes in testis cancer to determine the expression pattern in histologically distinct subtypes, correlate expression with genetic instability and correlate expression and genetic instability with clinical outcome. PATIENTS AND METHODS: 118 cases of testis cancer were analyzed. Immunohistochemical analysis of paraffin embedded specimens utilized monoclonal antibody for hMLH1 and hMSH2 mismatch repair proteins. Genetic instability was determined by comparing genomic DNA from microdissected matched normal and tumor cells. PCR amplification of 10 genetic markers assessed loss of heterozygosity and/or microsatellite instability. RESULTS: hMSH2 staining was associated with pathologic stage (p < 0.001) while hMLH1 staining was associated with cancer specific survival (p = 0.036). Genetic instability was detected in 94% of low hMLH1 and 92% of low hMSH2 staining tumors. Relapse and cancer specific death correlated with genetic instability (p = 0.01 and 0.04 respectively). Overall 9% of tumors exhibited reduced mismatch repair expression, microsatellite instability and an unfavorable clinical outcome. CONCLUSIONS: Mismatch repair expression and genetic instability define testis cancers with distinct molecular properties and clinical behavior. In conjunction with pathologic examination and serum tumor markers, mismatch repair expression may be an important determinant for clinical management of men with this malignancy. PMID- 15881920 TI - [Transvaginal urethrolysis for urethral obstruction after anti-incontinence surgery]. AB - PURPOSE: To analyse the success of transvaginal urethrolysis in resolving voiding dysfunction in patients following an anti-incontinence procedure. MATERIALS AND METHODS: A retrospective chart review was performed on 20 patients who had undergone transvaginal urethrolysis for urethral obstruction following an anti incontinence procedure. Preoperatively, a history was taken, pelvic examination and urodynamic were done. Mean patient age was 48 years, and the median time between the anti-incontinence procedure and the urethrolysis was nine months. Four patients had urinary retention and had irritative voiding symptoms. Previous surgery included pubovaginal sling in eleven patients, retropubic urethropexy in three and bladder neck suspension in six cases. Mean length of follow up after urethrolysis was 14 months. The urodynamic study demonstrated voiding flow rate of 9.9 ml/s and detrusor pressures at maximum flow of 48cmH20. RESULTS: Of the 20 patients 14 (70%) had relief of symptoms after a single urethrolysis, while two patients underwent a second transvaginal urethrolysis, with placement of a Martius flap between the urethra and the symphysis. There was no correlation between preoperative parameters examined and the outcome from urethrolysis. CONCLUSION: Our data support transvaginal urethrolysis for the treatment of urethral obstruction after anti-incontinence surgery. It is effective and minimally invasive technique with good results that should be considered if voiding dysfunction does not resolve spontaneously. PMID- 15881921 TI - [Surgical aspects in the third and fourth kidney retransplant]. AB - INTRODUCTION AND OBJECTIVES: A quarter of patients waiting for kidney transplantation are patients with previous graft failure. Outcome of first and second renal transplant make these the gold standard for end renal stage disease, but this is not so clear in the case of third and further renal transplant, especially at the time of organ shortage. We revise our experience in patients with three or more kidney transplants focusing on surgical aspects and graft outcome. MATERIAL AND METHOD: 1364 renal transplants have been carried out in our centre since 1975 until December 2003. We have retrospectively revised the 34 patients with three renal transplants and the 5 with four. We analyse the surgical technique, surgical complications and graft outcome. RESULTS: Mean age was 42 years (21-65). Average mismatches between donor and recipient was 3.2. All kidneys, but one case of living donor, were harvested from cadaver donors, mostly in multiple organ-procurement. Average time from the last renal transplant was 5 years (3 days-17 years) and from the last transplant carried out in the iliac fossa reused until the new transplant was 9 years (3 days- 17.5 years). All implants were performed through an iterative lumboliliac incision (25 on the right side, 11 on the left one and in 3 cases where side was not registered). Mean average duration of the procedure was 166 minutes (100-300). Nephrectomy of previous graft at the moment of the implant was carried out in 13 patients (33%). Vascular anastomosis was made on the common iliac vessels (50%) or on the external ones (50%) in end to side way, Ureteroneocystostomy was performed in an extravesical way except in 1 patient with cutaneous diversion. Vascular complications were 4 haemorrages (1 patient died), 3 venous and 2 arterial thrombosis. We had an abscess secondary to intestinal fistulae. Other surgical complications were 4 lymphoceles, three of them needed surgical treatment, and one perirenal haematoma treated in a conservative way. No urological complications were seen. In total 6 grafts (15%) were lost due to surgical complications. Graft actuarial survival rate at 1 year was 65%, 40% at 5 and 28% at 10 years. CONCLUSIONS: Three and four renal transplant survival rates are shorter than first and second ones. Iterative access through lumboiliac incision is associated with a higher vascular complication rate, probably in these patients a transperitoneal access would be better. Multicentric studies with higher numbers of patients are needed to define more clearly which patients would benefit from multiple kidney retransplants. PMID- 15881922 TI - [Correction of the penile curvature in ambulatory surgery]. AB - OBJECTIVE: To evaluate the results obtained in the correction of the curved penis by means of Nesbit's technique during the four first years of integration of our Service in the Major Ambulatory Surgery Unit (CMA) of our Hospital. PATIENTS AND METHOD: From January of 2000 to April of 2004 we intervened in ambulatory regime 21 patient suffering from curved penis (12 congenital and 9 with Peyronie's disease) by means of Nesbit's technique. The surgical-anesthetic performed procedure is described and also the criteria of inclusion and discharge are evaluated, as well as the results obtained and the degree of satisfaction by means of the elaboration of a questionnaire. RESULTS: None of the patients needed entrance for intrasurgery nor postsurgery complications. Thus, we obtained a null incidence of complications with the exception of the inevitable shortening of the penis, clearly independent from the regime of out-patient's process. The degree of satisfaction with the received treatment has been superior to 95%. CONCLUSIONS: The practical totality of the susceptible patients for surgical correction of penile curvature are candidates to be included in a CMA program, improving obviously the relation cost-efficacy, not diminishing for that reason the welfare quality nor the degree of patient's satisfaction. PMID- 15881923 TI - [Renal hydatidic cyst simulating hypernephroma]. AB - OBJECTIVE: To present a new case of hydatid cyst of the kidney with a difficult radiographic diagnosis. MATERIAL AND METHODS: We describe the clinical, diagnosis and treatment of a complex renal mass and its histological confirmation after surgery. Review of the literature. CONCLUSIONS: kidney's hydatidose is an unusual placement of this pathology. It is important to take care in the differential diagnosis in the context of complexes renal masses. There are some diagnosis procedures which can help us to establish it. Surgery is the treatment of choice in the majority of the cases. PMID- 15881924 TI - [Cystic ureteritis in a kidney transplantation candidate]. AB - Cystic ureteritis is a very uncommon pathology, whose pathogenesis is not well established. It is usually asociated with chronic infectious factors. It presents unspecific symptoms but characteristic radiologic findings. There is not an especific treatment for this disease. Kidney transplant is the final pathway for patients with chronic renal failure. We report a case of cystic ureteritis diagnosed during pre-transplant study. PMID- 15881925 TI - [Treatment of hemorrhagic cystitis secondary to cyclophosphamide. Literature review with regard to a case]. AB - There are extreme situations in the clinical treatment of intractable hematuria (fortunately not a common disease nowadays), in which even benign process may represent a serious risk to the life of the patient and where certain aggressive procedures like cystectomy have to be considered. Our objective in this paper is to review the different existing therapeutic alternatives for the treatment of an intense type of hematuria which does not yield to continuous saline serum irrigation. This study is partly based on a recent real clinical case in which, after the administration of cyclophosphamide to a young patient with recurrent and intense hematuria, the excretion was controlled through intravesical alum irrigation of thus avoiding cystectomy, a very aggressive surgical procedure with many after-effects. PMID- 15881926 TI - [Ureteral obstruction secondary to Rosai-Dorfman disease]. AB - Ureteral obstruction can be secondary to a lot of reason. We present a uncommon case report of ureteral obstruction to Rosai-Dorfman disease. The form of clinic presentation is described, the diagnostic methods used and the therapeutic management. PMID- 15881927 TI - [Cerebral metastasis of a pyelic urothelial carcinoma in a horsehoe kidney]. PMID- 15881928 TI - [Differential diagnosis in ureteral repletion defect]. PMID- 15881929 TI - [Bacterial colony as a complex community]. AB - Information on the adaptive behavior of cells in the process of the formation of a colony by bacteria Shigella flexneri Rd, used as a model, was obtained with the use of light, transmission and scanning microscopy. The process of the formation of a microcolony was demonstrated; at the initial stages (8 hours) it included 3 groups of cells, and 24 hours later it exhibited sharply defined structurization. The conclusion was made on the possibility of the joint use of different methods of microscopy for the study of the development of microcolonies and colonies of bacteria. PMID- 15881930 TI - [Influence of gaseous metabolites of soil bacteria on the multiplication of Listeria monocytogenes and Yersinia pseudotuberculosis]. AB - The influence of gaseous metabolites of saprophytic soil bacteria on the growth and multiplication of L. monocytogenes and Y. pseudotuberculosis was studied. The study revealed that all cultures under study exhibited selectively both inhibiting and stimulating action on the multiplication of test cultures. Bacteria of the genera Pseudomonas and Acinetobacter showed the highest inhibiting activitywith respect to the test cultures. Volatile metabolites of bacteria of the genus Aeromonas showed the highest stimulating activity. Methanol, according to the data of chromatographic analysis, supposedly played the main role in this process and was contained in the volatile substances of Aeromonas bacteria in greater amounts than in the volatile substances of Pseudomonas bacteria. PMID- 15881931 TI - [Antiadhesive effect of the secretion of the oral cavity in the system "Candida albicans--buccal epitheliocytes"]. AB - The influence of the oral cavity secretion on adhesive reactions in the system "C. albicans--buccal epitheliocytes" was studied. The treatment of C. albicans with natural saliva led to decrease of adhesive activity. The treatment of C. albicans with antibody absorbed saliva at different temperature conditions led to different changes of adhesion. This effect was determined by the action of temperature-dependent and temperature-independent factors, supposedly of enzymatic nature. PMID- 15881932 TI - [Cross-reacting antigens of pathogenic Burkholderia and some dangerous causative agents of infectious diseases]. AB - Cross-reacting antigens in B. mallei, B. pseudomallei, B. thailandensis, Francisella tularensis, Yersinia pestis and Mycobacterium tuberculosis were studied with the use of immuno- and electrophoretic techniques. The set of antigens was shown to be almost identical in the causative agents of glanders, melioidosis, as well as in B. thailandensis, though in the latter organism 200-kD glycoprotein was absent. The analysis of immuno- and proteinograms demonstrated the presence of cross-reactions in the representatives of the genus Burkholderia with the causative agents of plague, tularemia and tuberculosis, which served as the basis for making the scheme of their antigenic relationships. The use of immunosorption techniques with subsequent analysis of the preparations by means of the SDS polyacryl gel electrophoresis and immunoblotting made it possible to characterize cross-reacting antigens of the pathogenic microorganisms under study, to establish their molecular weights (81-15 kD) and to show that some detected antigens are analogous to B. pseudomallei outer membrane proteins (34 and 30 kD). PMID- 15881933 TI - [Specific clinical, epidemiological patterns and laboratory diagnostics of enterovirus infection in the Republic of Belarus]. AB - The clinical and epidemiological patterns as well as the results of the laboratory verification of the outbreak of enterovirus infection (EVI) in Minsk during the period of summer-autumn, 2000, are presented. During this outbreak a variety of clinical forms were observed, the serous meningitis being prevalent (57.5%). Practically simultaneous occurrence of infection on the territory of all administrative districts of the city, the predominant involvement of children aged up to 14 years into the outbreak, a high proportion of simultaneous casualities in the multiple foci. A number of circulating enteroviruses (EV)- ECHO 30, ECHO 6 of three serotypes and Coxsackie B5--were simultaneously isolated from clinical material. EV of the same serotypes were isolated from tap drinking water, and neutralizing antibodies to these serotypes were often detected in the patients blood sera. Infectious EV were also present in samples of bottled water and in water reservoirs used for bathing. The routes of EV transmission and the improvement of EVI control are discussed. PMID- 15881934 TI - [The occurrence rate of HGV/GBV-C RNA and risk factors in patients of narcological dispensary in Novosibirsk]. AB - The occurrence rate of HGV/GBV-C RNA, genotypic variety of isolates and various risk factors of infection with HGV/GBV-C were evaluated in 500 patients of the narcological dispensary of Novosibirsk. The occurrence rate of HGV/GBV-C RNA among all examined blood sera was 33.6%. At the same time in blood sera with HCV markers the occurrence rate of HGV/ GBV-C was 42.9% and in sera with negative results for markers HCV--25%. For gene typing of obtained isolates the direct sequencing of the amplification products of fragment NS3B and the phylogenetic analysis of the sequences thus obtained were used. Almost all isolates subjected to gene typing belonged to genotype 2, widespread in Europe, and only 1 isolate was classified with genotype 4. Statistically significant (p<0.05) risk of HGV/GBV-C infection among the examined subjects was linked with the intravenous use of drugs (OR 2.15), risky sexual behavior (OR 1.8) and the presence of virus hepatitis C (OR 2.26). PMID- 15881935 TI - [Genotypic emm-typing of streptococcal cultures, group A, isolated from patients with different clinical manifestations of streptococcal infection in Moscow]. AB - The results of the intraspecific typing of group A streptococci (SGA), isolated from patients with different manifestations of group A streptococcal infection, carried out in Russia for the first time, are presented. The genotypic method of emm-typing, based on sequencing the DNA area coding the variable part of the molecule of SGA M-protein was used. The data obtained in this study made it possible to follow changes in the pattern of SGA M-types in Moscow and the specific features of SGA in comparison with the analogous data on other territories. New emm-types of SGA circulating on the territory of Russia were detected and described. PMID- 15881936 TI - [Experimental evaluation of recombinant polypeptides as Streptococcus Group B vaccine]. AB - Recombinant polypeptides corresponding to the conservative N-terminal area of Bac surface protein of group B streptococci (GBS) and having the human IgA binding site were obtained and evaluated in terms of Immunogenic and protective properties. GBS strain 219, serotype 1 bc, served as the source of chromosomal DNA used for cloning. Three polypeptides P1, P5 and P6 were constructed. Polypeptides P1 and P5 with a molecular weight of 22 kD were coded by practically identical DNA fragments (685 nucleotide pairs) and differed in two aminoacids In their central part. Polypeptide P6 had a molecular weight of 35 kD, The fragment of recombinant Bac protein with a molecular weight of 35 kD was found to have protective properties: when used for the subcutaneous immunization of animals, it stimulated the development of immune response capable, in case of challenge, to induce accelerated elimination of streptococci and the prevention of the mice death. Recombinant fragment P6 of GBS protein Bac may be regarded as a candidate for inclusion into GBS-specific vaccine. PMID- 15881937 TI - [Evaluation of the immunogenic and reactogenic properties of vaccine "Euvax B" in persons aged 16-18 years in different geographical zones of Russia]. AB - The comparative evaluation of the reactogenic and immunogenic properties of vaccine "Euvax B" in adolescents aged 16-18 years was carried out in factory housing estate Lvovsky, the Podolsk district, the Moscow region, and in the Yamalo-Nenets Autonomous District (AD): in towns Salekhard, Noyabrsk, Muravlenko, in housing estate Tarko-Sale. The evaluation demonstrated that the use of this vaccine in a dose of 20 mg/ml produced no postvaccinal complications and the frequency of postvaccinal reactions was insignificant. Postvaccinal anti-HBs were detected, on the average, in protective concentrations in 98.4 +/- 1.8% of the vaccines (99.0 +/- 1.9% in the Moscow region, 97.7 +/- 3.2% in the Yamalo-Nenets AD). In most of the vaccinated adolescents high concentrations of postvacinal anti-HBs were detected: in 51.4 +/- 9.7% of the vaccinees in the Moscow region and in 89.7 +/- 6.4% in the Yamalo-Nenets AD they exceeded 1,000 I.U./lit. The conclusion was made that in the territories located in subarctic latitudes vaccinal prophylaxis against hepatitis B was safe and highly effective and have no limitations connected with the conditions of the Far North. PMID- 15881938 TI - [Immunovac-VP-4, immunomodulator decreases immunosuppression and enhances the cytotoxic activity induced by the cytostatic Cisplatin]. AB - The influence of the vaccine Immunovac-VP-4, prepared from the antigens of opportunistic microorganisms, on the proliferative and cytotoxic activity on peripheral blood mononuclears (PBMN) from healthy donors in vitro and on spleen cells of CBA mice in vivo during their incubation with Cisplatin was studied. VP 4 produced a dose-dependent, stimulating effect on the proliferative potential of PBMN and, when used in the highest of all tested doses (20 microg/ml), increased the Cisplatin-suppressed proliferative activity of PBMN in 9.4-fold. VP-4 increased the cytotoxic activity of PBMN on tumor line cells K-562 (38,4 to 60.1%) and increased the cytotoxic effect of Cisplatin (68.18 to 87.56%). A single injection of VP-4 to mice stimulated the proliferative activity of spleen cells, studied ex vivo, units and partially restored their cytostatic-suppressed activity. The cytotoxic action of the spleen cells of immunized mice on tumor line cells YAC-1 was twice as great as that of spleen cells taken from intact animals and potentiated the cytotoxic action of Cisplatin. The mechanism of increasing the proliferative activity and cytotoxic effect of monomuclears under the influence of vaccine VP-4 is seemingly linked with the synthesis of cytokines, influencing the lymphokine-activated cytotoxicity of lymphocytes. PMID- 15881939 TI - [Apoptosis of phagocytes as one of the probable mechanisms of the pathogenetic action of Yersinia pestis "mouse" toxin]. AB - Apoptosis was evaluated by characteristic morphological changes of cells in preparations stained with histological dyes and in live preparations, as well as by DNA degradation, colorimetrically detected with the use of the diphenylamine reagent. "Mouse toxin" (MT) was found to have a pronounced apoptogenic action with respect to the phagocytic cells of mice, but not guinea pigs. Macrophages were affected by this action stronger than neutrophils, and in both cases this effect was dose dependent. As the dose of MT decreased to 0.01 microg/ml, the proportion of cells dying as the result of apoptosis increased, the necrotic type of damage was almost absent. On the contrary, as MT concentration rose to 1.0 microg/ml and over, the proportion of phagocytes dying due to necrosis increased with a decrease in the number of cells in which the process of apoptosis started. The results of the study are indicative of the fact that the mechanisms programming the death of cells under the action of MT on macrophages and neutrophils took part in the process, which, in its turn, determined their role in the pathogenesis of plague. PMID- 15881940 TI - [The evaluation of the diagnostic value of PCR test systems according to the results of state trials]. AB - The method for the diagnostic value evaluation of preparations, based on the calculation of the accuracy of test results and taking into account the spread of the diagnosed disease (the comparison of the PCR test systems for the diagnostics of hepatitis B and ureaplasmosis) is proposed. As shown in this work, evaluations obtained with the use of this method coincide with those obtained on the basis of prognostic value, but are more convenient in use and provide additional information. PMID- 15881941 TI - [Relationships between the probiotic and host indigenous lactobacilli under the conditions of mixed cultivation in vitro]. AB - Relationships between 5 Lactobacillus manufacturing strains, 458 cultures of indigenous lactobacilli isolated from the human digestive and vaginal tracts and 98 isolates from the feces of white rats and mice were under study. The study demonstrated that under the conditions of mixed in vitro cultivation of paired cultures, probiotic strains inhibited more than 60% of the indigenous lactobacilli isolates. L. acidophilus strain K3 III 24 had the widest spectrum of antagonistic activity. Antagonistic relationships between indigenous lactobacilli depended on the origin, individual features and the anatomical sites of the culture isolation. Based on these results it has to be suggested that probiotic lactobacilli are capable of inducing disbalance in the host indigenous lactoflora. While choosing probiotics the character of relationships between probiotic microorganisms and the indigenous lactobacilli of the future recipient is recommended to be preliminarily tested in vitro. PMID- 15881942 TI - [Comparative characterization of the microflora of the upper respiratory tract mucous membranes in bronchial asthma and allergic persistent rhinitis]. AB - A total of 250 patients with diagnosed bronchial asthma (BA) were examined by microbiological methods. Among them--188 children and 62 adults. In 87 patients the microflora of nasal mucosa was studied, in 40--of pharynx only and in 123 patients--both the nasal and the pharynx. For comparative analysis earlier data obtained in 69 patients with persistent allergic rhinitis (PAR) were used. The cultures isolated from the nasal mucosa of BA patients were shown to number 18 genera and 42 species, while among those isolated from pharynx mucosa 20 genera and 40 species. Monocultures were isolated from the nasal mucosa only in 23% of the examined patients and from the pharynx mucosa--only in 1.42%. Associations with different numbers of components were isolated from nasal and pharynx mucosa (2 to 6, 2 to 8 respectively). Staphylococcus aureus was regarded as the main species of nasal biocenosis in BA and PAR, as well as pharynx biocenosis in BA. Besides, in BA other Staphylococcus species (schleiferi, caprae, capitis, hominis, etc.), reversely related to the main species, could be isolated from both mucous membranes. Similarities and differences in microflora of biocenoses in both nosological forms, confirming links between PAR and BA, are considered. PMID- 15881943 TI - [Association of Candida albicans fungi with some opportunistic microorganisms in intestinal dysbiosis in patients of different age groups]. AB - The occurrence of C. albicans in association with opportunistic microorganisms (Staphylococcus aureus and Klebsiella) in intestinal dysbiosis in patents of different age groups (from 6 days to 31 years) was analyzed. The data on the comparative evaluation of the results of the bacteriological examination of patients with intestinal dysbiosis, carried out during the periods of 2000-2001 and 2001-2002 (388 and 467 patients respectively), are presented. During the period of 2000-2001 the detection rate of C. albicans in monoculture was 6-33% of cases (in the examined group). A higher detection rate was registered with respect to the association of C. albicans with staphylococci (42-65% of cases). The associations of C. albicans with staphylococci and Klebsiella were observed in 13-46% of cases. Similar results were registered during the period of 2001 2002, but during this period a reliable decrease in the detection rate of the association of C. albicans with staphylococci (practically by 20% in the group of infants) and the 1 1/2-fold increase of the detection rate of C. albicans in monooulture (in all age groups) were observed. PMID- 15881944 TI - [Capacity of fusicoccin for inducing the production of early interferon]. AB - The response of human and animal cells to the action of fusicoccyne (FC), a fungal metabolite with phytohormonal properties, was evaluated. As shown by in vitro studies, FC had the capacity to induce the production of early interferon (IFN) in the blood serum of non-inbred white mice and to enhance the natural cytotoxic activity of human lymphocytes. In vitro experiments also revealed that the action of FC inhibited the metabolism of actively proliferating monocytic leukemia cells J-96 and human ovarian carcinoma cells CaOv, as well as mouse fibroblasts L-929. The common character of the mechanism of action of FC and IFN, having well-known antiproliferative and immunomodulating activity, is discussed. PMID- 15881945 TI - [Antibiotic resistance of shigellae and rationale for etiotropic therapy of Shigella infections]. AB - The study was aimed at determining sensitivity of shigellae to antibacterial preparations and their clinical effectiveness for correcting recommendations on the empirical therapy of acute Shigella infections (ASI). The sensitivity of 164 S. flexneri strains and 80 S. sonnei strains, isolated in 1996-2003 in the Sumy region, Ukraine, was determined with respect to 19 antibacterial preparations: ampicillin (Am), tetracycline (Te), rifampicin (Ri), chloramphenicol (Ca), streptomycin (St), fusidin (Fu), kanamycin (Kn), erythromycin (Er), carbenicillin (Cb), doxycycline (Do), gentamicin (Ge), ofloxacin (Of), cefazolin (Cf), ciprofloxacin (Cp). S. flexneri and S. sonnei were found to be highly sensitive to Am (100%), Te (100%), Cb (90% and 50% respectively), Do (90% and 35% respectively), Fu (100%), Er (100%), Ri (100%), Ca (71.8% and 45% respectively), St (81% and 40% respectively). Some isolated cultures were resistant to fluorochinolones. In addition, the clinical and laboratory analysis of the effectiveness of some preparations was carried out. A total of 202 patients, divided into 6 groups, received furazolidone, chloramphenicol, norfloxacin, phthalazole, polymyxin and the combination of several antibacterial preparations. High efficiency of norfloxacin in the treatment of ASI was confirmed. The use of other preparations and their combinations was found to produce only a slight effect. PMID- 15881946 TI - [Complex typing of Salmonella typhimurium hospital strains]. AB - A salmonellosis outbreak, caused by S. typhimurium, was investigated with the use of some microbiological and molecular-biological methods of typing. This investigation revealed that the outbreak was caused by the "outbreak" electrotype of the multi-resistant variant of the infective agent, found to have several plasmidovars. The possibilities and limitations of typing by sensitivity to antibiotics and plasmid DNA profile were shown. These methods of intraspecific typing were regarded as methods making it possible to establish the heterogeneity of S. typhimurium with the use of intraclonal markers. PMID- 15881947 TI - [Yersinia pseudotuberculosis nucleoside-kinase]. AB - Enzyme capable of catalyzing the phosphorylation of thymidine and uridine was isolated from Y. pseudotuberculosis cells by fractionation with the use of ammonium sulfate, ion exchange and affinity chromatography. The degree of purification of thymidine- and uridine-kinase was approximately 350 times, and at all stages of isolation the activity of both nucleoside-kinases was detected in the same peaks. The purified enzyme was capable of the phosphorylation of thymidine and uridine at temperatures of 8-10 degrees C to 50 degrees C and exhibited the maximum enzymatic activity at pH 8-8.5 and 45 degrees C in the presence of 0.5-1.0 mM MgCl2 and 2 mM ATP. The enzyme was found to have no strict substrate specificity and transferred the phosphate group from ATP to radiolabeled thymidine, uridine and desoxycytidine with different effectiveness, but did not use thymidine-monophosphate as phosphate acceptor. PMID- 15881948 TI - [Intensity of the production of rheumatoid factor in patients with different degrees of sensitization to Borrelia garinii antigens]. AB - The content of rheumatoid factor and the levels of IgA, IgM, IgG in patients with different degrees of sensitization to Borrelia garinii antigens were studied. The titer of rheumatoid factor was found to increase in accordance with the growth of the levels of anti-Borrelia antibodies. A rise in the concentration of serum immunoglobulins simultaneously with the development of Borrelia infection was registered. The mechanisms of autoimmunization are discussed, taking into account the known biological properties of the causative agent of Lyme disease. PMID- 15881949 TI - [Clinical and immunological effect produced by vaccination with "Pneumo 23" of children with atopic bronchial asthma]. AB - A total of 45 children aged 4-16 years with atopic bronchial asthma of different severity level-resulted from a past case of outhospital pneumonia, received vaccine "Pneumo 23" ("Aventis Pasteur", France). The vaccine was found to be well tolerated, no cases of the exacerbation of the atopic process were registered. A reliable Increased level of specific antibodies to the mixture of polysaccharides, contained in the vaccine "Pneumo 23", was found to occur. The tendency towards decreased level of serum IgE was established. PMID- 15881950 TI - [Characterization of Escherichia coli donor-specific RNA-containing bacteriophages]. AB - The biological properties of 9 clones of Ri bacteriophages isolated from sewage water in 1981 were studied. On the basis of the activity of Ri phages with respect to E. coli donor-specific strains K12, the type of negative colonies, the ultrastructure of the virion and its sizes, adsorption on the pili of host cells, the latent period, the amount of harvest obtained from one infected cell, the clones under study were classified with small spherical RNA-bacteriophages. The neutralization of Ri phages with antiphage sera to standard phages f2 and fr made it possible to classify them with the first serological group and to divide them into 3 subgroups. PMID- 15881951 TI - [Dysbacteriosis and immunopathological process]. AB - The poststress translocation of intestinal microflora to the internal environment of the body is proposed to be regarded as the mechanism of neuroimmune cooperation in the realization of adaptative raction. It is pointed out that the effectiveness of the adaptive process depends on the state of microbiocenosis as a mediating factor. The inclusion of remedies for the correction of intestinal microbiocenosis into the complex therapy of patients with chronic pathology (as a specific manifestation of dysadaptive state) makes it possible to achieve results which cannot be achieved by the standard approach. We should return to the initial interpretation of I.I.Mechnikov's thesis on the basic role of dysbacteriosis in the development of the pathological process. PMID- 15881952 TI - [Development of antileprosy vaccines]. AB - The absence of an effective antileprosy vaccine, capable of preventing the spread of leprosy hinders its control in endemic countries. Developing such a vaccine is highly difficult due to the absence of reproducible methods for the in vitro cultivation of Mycobacterium leprae. The results of field trials of earlier proposed vaccines (BCG and BCG in combination with killed M. leprae) are indicative of their insufficient efficacy. The article presents a review of literature, including historical information, current problems and the main approaches to the development of vaccine against leprosy. PMID- 15881953 TI - [Characterization of Bacillus anthracis typical virulent test strain 81/1]. AB - The results of the prolonged and many-sided study of B. anthracis strain 81/1 by different authors are presented. The cultural and morphological, biochemical, antigenic, molecular-genetic characteristics of this strain give grounds for regarding it as a typical test strain to be used for the determination of the vaccines immunogenicity, the effectiveness of antibiotics and immunomodulators. PMID- 15881954 TI - [Resistance to a causative agent of infection and immune response]. AB - An analytical literature review dealing with the problems of resistance to causative agents of infections. The review includes the data of experimental research, as well as the author's own immunological studies in children at the clinic of acute infections. Mechanisms of resistance at different phases of the infectious process, the multivalued role of Th1- and Th2-dependent responses in different infections, the role of vaccinal immunity in the resistance of children to the causative agents of vaccine-preventable infections (diphtheria and parotitis) are analyzed. PMID- 15881955 TI - [Epidemiology of tick-borne encephalitis in the Kirov region]. PMID- 15881956 TI - [Sewage waters as an indicator of the viral circulation among population]. PMID- 15881957 TI - [Investigation on mixed natural foci of zoonotic infections]. PMID- 15881958 TI - [Sanitary and epidemiological masterpieces of the century before last (face to face with history)]. PMID- 15881959 TI - [Clinical and epidemiological characteristics of tuberculosis in children and adolescents in Armenia]. AB - In Armenia, the incidence of tuberculosis among children aged less than 18 years increased by 53.2% (from 7.9 to 12.1% per 100,000 inhabitants) in 1993 to 2003. On the contrary, the proportion of children and adolescents among all the patients decreased by almost twice (from 13.4 to 8.1%), which is largely due to the reduction in birth rates and to the inadequate detection of minor and local forms of primary tuberculosis. The clinical composition of tuberculosis in 1270 examined patients was the following: tuberculosis of respiratory organs (82.9%); its extrapulmonary forms (14.4%); tuberculous intoxication (2.7%); a concomitance of pulmonary and extrapulmonary tuberculosis (5%). The boy-girl ratio was 2:1. Tuberculosis of intrathoracic lymph nodes was predominant in children aged less than 14 years; infiltrative tuberculosis was prevalent in adolescents. Lung tissue destruction was observed in 15.7%; Mycobacterium tuberculosis was found in 10.2% of the patients. 49.9% of the patients were from the foci of tuberculosis; half of these cases were from the hidden foci of this infection. PMID- 15881960 TI - [Clinical analysis of reasons for the absence of bacterial isolation in new cases of destructive pulmonary tuberculosis]. AB - Saint Petersburg has been marked by a reduction in the efficiency of Mycobacterium tuberculosis (MBT) detection in the past 5 years. This is suggested by negative changes in the ratio of the rate of detected bacterial isolation to the phase of lung decay in new cases of tuberculosis. This is largely due to the fact that before treatment, clinicians do not fulfil in full measure the regulated requirements for the choice of studies and for the frequency of examinations for MBT. Sampling for microbiological studies is made, by infringing the technological specifications. Highly informative methods for MBT detection are not frequently used in the above cases. This all hampers diagnosis and rational treatment in these patients. PMID- 15881961 TI - [Computed tomography at the outpatient stage of differential diagnosis of destructive inflammations and tuberculosis of the lung]. AB - The authors retrospectively analyzed the quality of RCT differentiation of destructive lung lesions at outpatient diagnostic units of multidisciplinary polyclinics. At the first stage, the recognition accuracy rate was not more than 73%, which is attributable to the high macroscopic similarity of bacterial destructive inflammations and specific tuberculous processes. Both conditions were frequently complicated by the addition of fungal invasion that made their differentiation all the more difficult. It has found that during the tense epidemiological situation associated with tuberculosis, there is its explainable, but not always justified hyperdiagnosis. The ratio of false-negative to false positive tuberculosis results was 1:2. The paper is illustrated by different types of destructive inflammations that mimic tuberculosis and by the trend of some of the types is shown. Emphasis is laid on the neglects and errors that cause diagnostic differences. A half of them is attributable to the objective complexities of its diagnosis; the other (subjective) half is a potential reserve for upgrading the quality of differential diagnosis in phthisiopulmonology. PMID- 15881962 TI - [Comparative efficiency of treatment with moxyfloxacin and lomefloxacin for generalized murine tuberculosis caused by drug-resistant Mycobacterium strains]. AB - The antituberculous efficacy of the new fluoroquinolone agent moxyfloxacin was studied on a model of generalized murine tuberculosis caused by a multidrug resistant Mycobacterium strain. There was evidence for the high therapeutic effect of moxyfloxacin and its advantages over the antituberculous agent lomefloxacin. PMID- 15881963 TI - [Current ways of preventing postresectional bronchial fistulas in pulmonary tuberculosis]. AB - The paper presents the data of a follow-up of 51 patients in whom a bronchial stump suture was additionally fixed at surgery by applying Tochocomb plates (in 30 patients) or an autoangiograft from the removed lung (in 21 patients) in order to prevent a bronchial fistula. The preliminary findings were positively appraised. The authors recommend that this procedure be used in phthisiosurgery and surgical pulmonology. PMID- 15881964 TI - [Surgical treatment of bilateral pulmonary echinococciasis in active tuberculosis]. PMID- 15881965 TI - [The blood oxygen-transport system and oxygen tissue balance in patients with bronchial asthma (BA) concurrent with carbohydrate metabolic disturbances]. AB - Examining the parameters of the blood oxygen-transport system and oxygen tissue balance in 71 patients with bronchial asthma (BA) and in 60 patients with BA concurrent with varying carbohydrate metabolic disturbances (CMDs) revealed differences in oxygen transport. In asthmatic patients without CMDs, the pattern of changes in oxygen balance permitted identification of arteriohypoxemic hypoxia. The high frequency of factors (alkalosis, hypocapnia) that enhance hemoglobin affinity for oxygen, which are compensated for by the increase in 2,3 DPH, as well as hypoxia of peripheral shunting are the specific features of impaired oxygen balance in asthmatic patients with concomitant CMDs that predispose to the development and progression of pulmonary hypertension and cor pulmonale which cause early disability. The determination of the leading link in impaired oxygen transport individualizes treatment approaches and provides evidence for a need for earlier use of metabolic and antihypoxic agents in asthmatic patients with CMDs. PMID- 15881966 TI - [Differential diagnosis of specific and nonspecific lung diseases by the parameters of the leukocytic enzymatic system]. AB - The study examined 56 patients who were divided into three groups: 1) patients with infiltrative pulmonary tuberculosis; 2) those with nonspecific pneumonia; 3) control (apparently healthy individuals). In addition to the routine studies, the R. P. Nartsissov procedure was used to determine the activity of SDH and alpha GPDH in all the examinees. The findings indicate that the lymphocytic activity of basic energy processes is much decreased in infiltrative tuberculosis and nonspecific pneumonia. Of the greatest value for active diagnosis is the determination of alpha-GPDH activity that is reduced only in patients with infiltrative pulmonary tuberculosis, its low activity points to profound redox disorders. PMID- 15881967 TI - [Role of membranous disturbances in the development of bronchial hyperresponsiveness]. AB - The biophysical parameters of lymphocytic membranes and their phospholipid spectrum were studied in relation to the manifestations of nonspecific hyperreactivity of the bronchial tree in 69 patients with bronchial asthma of varying severity. Impairments of the biochemical and structural states of membranes were ascertained to be one of the major pathogenetic mechanisms responsible for the clinical, pathophysiological, and pathomorphological manifestations of the disease, its severity and persistent course. PMID- 15881968 TI - [Correlation between the expression of Rv3286c, Rv2626c, Rv2031c, AND Rv3133c and the tolerance of Mycobacterium bovis BCG to rifampicin and metronidazole in different physiological conditions]. AB - The fact that antibiotics exert no effect on Mycobacterium tuberculosis may be attributable to the production of their persistent forms. A correlation was studied between the expression of the genes Rv3286c, Rv2626c, Rv2031, and Rv3133c and the phenotypical tolerance to antibiotics in relation to the physiological condition of M. bovis BCG. The anaerobic growth conditions ("Wayne dormancy model") were found to be attended by the increased expression of all the studied genes, by the higher tolerance to rifampicin, and by the decreased phenotypical resistance to metronidazole. PMID- 15881969 TI - [Minutes of the 238th meeting of the Section of Thoracic Surgery of the Surgery Society of Moscow and the Moscow Region on April 27, 2004]. PMID- 15881970 TI - [Total endoprosthetic repair of the knee joint in patients with sequelae of tuberculous gonitis]. PMID- 15881971 TI - [Latent tuberculosis infection]. PMID- 15881972 TI - National exposure measurements for decisions to protect public health from environmental exposures. AB - Protecting public health from environmental exposures requires four steps: detection of exposures known or expected to cause disease, assessment of health risk from exposure, implementation of an exposure intervention, and assurance that the exposure intervention is effective. To prioritize efforts in these four areas one must consider the size of the population affected, the seriousness of health effects, and the availability of cost-effective exposure interventions. Population exposure data is critical to each of these steps for protecting health. Biomonitoring data for the US population is now available to assist public health scientists and physicians in preventing disease from environmental exposures, and it complements that available for levels of chemicals in environmental media. The Second National Report on Human Exposure to Environmental Chemicals provides for the US population serum, blood and urine levels for 116 environmental chemicals over the years 1999 and 2000, with separate analyses by age, sex, and race/ethnicity. This national exposure information identifies which chemicals get into Americans in measurable quantities; determines whether exposure levels are higher among population subgroups; determines how many Americans have levels of chemicals above recognized health threshold levels (for chemicals with such threshold levels); establishes reference ranges that define general population exposure so unusual exposures can be recognized; assesses the effectiveness of public health efforts to reduce population exposure to selected chemicals; and tracks over time trends in US population exposure. Blood lead measurements in the population were important in identifying lead in gasoline as a significant source of human lead exposure and documenting the reduction in blood lead levels in the population as a result of removing lead from gasoline and other products in the United States. Serum cotinine levels in the early 1990s found more widespread exposure to environmental tobacco smoke (ETS) in the United States than previously thought and additional measurements in 1999 and 2000 documented major declines in exposure to ETS as a result of public health actions in the 1990s. A new biomonitoring assessment of the exposure of the US population will be released every 2 years as the "National Report on Human Exposure to Environmental Chemicals." These reports will include the current 116 chemicals and new chemicals added to monitor priority exposures of the population. PMID- 15881973 TI - Human health research and policy development: experience in the Great Lakes region. AB - As a direct outgrowth of industrial and agricultural activities, the quality of the Great Lakes ecosystem has declined significantly because of toxic substances in the water, eutrophication, overfishing, and invasive species that have been introduced into the waterways. Although measures have been adopted to restore the health of the ecosystem, contamination of Great Lakes sport fish continues arising from conditions that still prevail, but on a more limited scale. As a consequence, the Great Lakes states have issued guidelines for the public in the form of health advisories for fish consumption to encourage practices that will minimize exposure to contaminants found in Great Lakes sport fish. Scientific research has strongly influenced many policy decisions, including the development of laws, rules, and guidelines applicable to public health not only in regard to fish advisories but also other issues impacting human health. This paper proposes to outline how policy has been influenced by scientific findings and the far reaching effect that these decisions have had on the health status of the public in the Great Lakes area and its potential for influencing the nation as a whole and our global neighbors. Within the Great Lakes basin, polychlorinated biphenyls (PCB) and mercury are the subject of the greatest number of fish advisories. Great Lakes-based researchers have studied populations residing in the Great Lakes basin to determine their level of awareness concerning fish consumption health advisories. They found that almost 50% of the residents who consumed Great Lakes sport fish were aware of sport fish consumption advisories. Of those with awareness, almost 60% were males and only about 40% were females. The researchers attributed the greater awareness among males to the health advisory materials that males receive with their fishing licenses and to their contact with fishing related groups. The lower level of awareness among women regarding fish consumption advisories subsequently prompted the researchers to recommend targeting risk communication programs for female consumers of Great Lakes sport fish, particularly women of reproductive age. The Wisconsin Department of Health and Family Services subsequently followed the recommendation and developed uniform outreach materials for women, minorities, and the general public to be used by the Great Lakes states. The policy change directing educational materials to at-risk groups (e.g., women of reproductive age and minorities) is a direct outgrowth of the finding of low awareness about fish advisories among women who were interviewed. PMID- 15881974 TI - Improving strategies to prevent childhood lead poisoning using local data. AB - Lead poisoning remains an important, yet entirely preventable, disease among children worldwide. Children's blood lead levels (BLLs) have been declining in the United States; however, nearly half a million children have BLLs > or = 10 microg/ dl, the level targeted for elimination by 2010. Attainment of this national goal will require translating knowledge into public health practice. The Centers for Disease Control and Prevention (CDC) funds state and local health departments to develop comprehensive prevention programs and surveillance. The Jefferson County, Kentucky Program, which includes Louisville, adopted CDC's recommendation for targeting lead testing to children at highest risk and used knowledge of risk factors for lead poisoning to develop prevention strategies. Blood lead testing was targeted to Louisville neighborhoods at high risk, i.e., those characterized by housing built before 1950 and valued < $50,000, which are known risk factors for BLLs > or = 10 microg/dl among children. We evaluated the impact of these and other interventions. Testing of children aged 9-24 months who were born in high risk housing increased from 64.5% to 73.7% (p-value <0.001) among the 1996 and 2000 birth cohorts. Among the 1996 and 2000 birth cohorts, there was no significant change in testing of children born in low risk housing, i.e., built after 1950 and valued at > or = $50,000 (37.0-37.5%; p-value = 0.649). This report demonstrates that applying scientific knowledge to public health practice and using surveillance and other data to evaluate practice effectively increased testing of children at high risk for lead poisoning, increased lead-safe housing, and empowered communities to protect their children from lead exposure. PMID- 15881975 TI - Ensuring a scientific basis for community interventions for asthma. AB - Community based interventions are an important part of public health management of many diseases, including asthma. However, there are few scientifically proven and readily available community interventions for asthma. In an effort to increase the number of available interventions, we have identified ongoing asthma intervention research, identified potentially effective asthma interventions based on completed research, and prepared several of the effective interventions for widespread implementation through a process called "translation." We provide an example of one of these effective interventions now available for widespread implementation, "Creating a medical home for asthma." This intervention grew out of need for an intervention in New York City Department of Health (NYCDOH) clinics. The intervention includes training all clinic staff in a comprehensive, preventive approach to asthma care. All of the materials needed to implement the intervention are available to all through the NYCDOH web site (www.nyc.gov/ html/doh/html/cmha/index.html). This example points to the importance of making the tools needed to implement effective interventions available across the country and the role of public/private partnerships to assure the availability of science-based interventions for asthma control. PMID- 15881976 TI - Food service workers' self-reported food preparation practices: an EHS-Net study. AB - This study was conducted by the Environmental Health Specialists Network (EHS Net), a network of environmental health specialists and epidemiologists at federal and state health agencies, whose mission is to improve environmental health practice. One of EHS-Net's primary goals is to improve the understanding of the underlying causes of foodborne illness using a system-based approach. As part of this ongoing effort, EHS-Net analyzed data from a telephone survey of food service workers designed to increase our understanding of food preparation practices (a cause of foodborne illness) in restaurants. Results indicated that risky food preparation practices were commonly reported. Respondents said that at work they did not always wear gloves while touching ready-to-eat (RTE) food (60%), did not always wash their hands or change their gloves between handling raw meat and RTE food (23% and 33%), did not use a thermometer to check food temperatures (53%), and had worked while sick with vomiting or diarrhea (5%). Several factors were associated with safer food preparation practices. Workers responsible for food preparation reported washing their hands and wearing gloves when handling RTE food more often than workers not responsible for food preparation. Workers who cooked reported changing their gloves more often than workers who did not cook. Older workers and managers reported washing their hands more often than younger workers and non-managers. Workers in chain restaurants more frequently reported using thermometers than workers in independently owned restaurants. This study provides valuable information concerning the prevalence of food preparation practices and factors that may impact those practices. Additional research is needed to better understand those factors. PMID- 15881977 TI - Surveillance of hazardous substance emergency events: identifying areas for public health prevention. AB - The Hazardous Substances Emergency Events Surveillance (HSEES) system is a comprehensive, state-based surveillance system of hazardous substance releases and public health consequences. Maintained by the Agency for Toxic Substances and Disease Registry (ATSDR) since 1990, the system captures information on acute releases of hazardous substances that need to be cleaned up or neutralized according to federal, state, or local law. Information about threatened releases that result in public health action such as evacuation is also included. Of the 39,766 events reported to HSEES for 1996--2001, 8% resulted in deaths or injuries. Funded through a competitive program announcement, 15 states currently participate in HSEES. State coordinators actively collect data from multiple sources after an eligible event occurs and enter data about the event into a standardized ATSDR-provided web-based system. The information in HSEES describes the distribution and characteristics of hazardous substances emergencies and the morbidity and mortality experienced by employees, responders, and the general public as the result of hazardous substances releases. Analysis of HSEES data helps identify risk factors associated with hazardous substances releases. For example, although events in which chlorine was released account for only 1.6% of all events, they were 3.52 times more likely to result in injuries. Knowledge of these factors is useful in planning public safety interventions and can impact the formulation of guidelines and policies to help reduce the number of events (primary prevention) and the morbidity and mortality associated with these events (secondary prevention). Utilizing state-specific analyses of HSEES data, participating states have been able to develop prevention outreach activities such as awareness training of first responders, primary prevention of spills, and secondary prevention of related injuries and deaths caused by ammonia, chlorine, and mercury. Specific examples involving ammonia, chlorine, and mercury releases will be presented in detail. PMID- 15881978 TI - Public health partnerships addressing childhood cancer investigations: case study of Toms River, Dover Township, New Jersey, USA. AB - Toms River, located in Dover Township, Ocean County, New Jersey, USA, experienced an increased incidence in childhood leukemia, brain, and central nervous system cancers from the mid-1980s through the early 1990s. These findings initiated a series of community-based activities that lead to the establishment of a successful partnership between the community, public health, and environmental agencies. The common goal of this partnership was to investigate linkages between environmental exposures and childhood cancers. The investigation was comprehensive in nature and a product of an extensive collaborative effort on the part of community, local, state, and federal health agencies, and university research organizations. Central to the success of this partnership was development of a public health response plan. This response plan served to coordinate activities of various entities and ensure that actions to cease or reduce ongoing exposures were implemented in addressing the incidence of childhood cancers using the partnership paradigm. The authors propose six rules of engagement: (1) seek out willing participants, (2) establish an equitable partnership, (3) consider each partner's perspective, (4) define goals and roles for each partner, (5) seek out innovative opportunities, and (6) assure scientific credibility. The application of these rules of engagement led to innovations and advances in the fields of environmental health science and public health practice. PMID- 15881979 TI - Exposure pathway evaluations for sites that processed asbestos-contaminated vermiculite. AB - The Agency for Toxic Substances and Disease Registry (ATSDR) is currently evaluating the potential public health impacts associated with the processing of asbestos-contaminated vermiculite at various facilities around the country. Vermiculite ore contaminated with significant levels of asbestos was mined and milled in Libby, Montana, from the early 1920s until 1990. The majority of the Libby ore was then shipped to processing facilities for exfoliation. ATSDR initiated the National Asbestos Exposure Review (NAER) to identify and evaluate exposure pathways associated with these processing facilities. This manuscript details ATSDR's phased approach in addressing exposure potential around these sites. As this is an ongoing project, only the results from a selected set of completed site analyses are presented. Historical occupational exposures are the most significant exposure pathway for the site evaluations completed to date. Former workers also probably brought asbestos fibers home on their clothing, shoes, and hair, and their household contacts may have been exposed. Currently, most site-related worker and community exposure pathways have been eliminated. One community exposure pathway of indeterminate significance is the current exposure of individuals through direct contact with waste rock brought home for personal use as fill material, driveway surfacing, or soil amendment. Trace levels of asbestos are present in soil at many of the sites and buried waste rock has been discovered at a few sites; therefore, future worker and community exposure associated with disturbing on-site soil during construction or redevelopment at these sites is also a potential exposure pathway. PMID- 15881980 TI - Use of a systems-based approach to an environmental health assessment for a waterborne disease outbreak investigation at a snowmobile lodge in Wyoming. AB - Investigations into disease outbreaks generally incorporate an epidemiologic investigation, laboratory analysis, and an environmental health assessment. This last component is designed to discover connections between factors in the environment and the outbreak, but is often limited, either by time and resources, or the expertise of the personnel included in outbreak investigation teams. A waterborne Norovirus outbreak investigation in Sheridan County, Wyoming, in 2001 provides an excellent example of the importance of including an in-depth, systems based environmental health assessment in outbreak investigations. The epidemiologic component of this investigation identified the water supply of a snowmobile lodge in the Bighorn Mountains as the source of the outbreak, a result that was confirmed by laboratory analysis. Including a systems-based environmental health assessment in this investigation also helped to uncover the underlying environmental factors that led to contamination of the water supply. Those factors included an onsite wastewater disposal system that was overloaded by increased use and not well suited to local soil and geologic conditions and a drinking water system with no treatment or disinfection. In addition, heavy precipitation and increased pumping of wells to satisfy higher demands probably facilitated the contamination of the drinking water wells by causing greater movement of wastewater through the soil and underlying bedrock. By focusing on these links between factors in the environment and adverse health outcomes, the systems-based environmental health assessment also helped to highlight prevention strategies for avoiding recurrences. PMID- 15881981 TI - Cardiorespiratory hospitalizations associated with smoke exposure during the 1997, Southeast Asian forest fires. AB - We investigated the cardiorespiratory health effects of smoke exposure from the 1997 Southeast Asian Forest Fires among persons who were hospitalized in the region of Kuching, Malaysia. We selected admissions to seven hospitals in the Kuching region from a database of all hospital admissions in the state of Sarawak during January 1, 1995 and December 31, 1998. For several cardiorespiratory disease classifications we used Holt-Winters time-series analyses to determine whether the total number of monthly hospitalizations during the forest fire period (August 1 to October 31, 1997), or post-fire period (November 1, 1997 to December 31, 1997) exceeded forecasted estimates established from a historical baseline period of January 1, 1995 to July 31, 1997. We also identified age specific cohorts of persons whose members were admitted for specific cardiorespiratory problems during January 1 to July 31 of each year (1995--1997). We compared Kaplan-Meier survival curves of time to first readmission for the 1997 cohorts (exposed to the forest fire smoke) with the survival curves for the 1995 and 1996 cohorts (not exposed, pre-fire cohorts). The time-series analyses indicated that statistically significant fire-related increases were observed in respiratory hospitalizations, specifically those for chronic obstructive pulmonary disease (COPD) and asthma. The survival analyses indicated that persons over age 65 years with previous hospital admissions for any cause (chi2(1df) = 5.98, p = 0.015), any cardiorespiratory disease (chi2(1df) = 5.3, p = 0.02), any respiratory disease (chi2(1df) = 7.8, p = 0.005), or COPD (chi2(1df) = 3.9, p = 0.047), were significantly more likely to be rehospitalized during the follow-up period in 1997 than during the follow-up periods in the pre-fire years of 1995 or 1996. The survival functions of the exposed cohorts resumed similar trajectories to unexposed cohorts during the post-fire period of November 1, 1997 to December 31, 1998. Communities exposed to forest fire smoke during the Southeast Asian forest fires of 1997 experienced short-term increases in cardiorespiratory hospitalizations. When an air quality emergency is anticipated, persons over age 65 with histories of respiratory hospitalizations should be preidentified from existing hospitalization records and given priority access to interventions. PMID- 15881982 TI - The usefulness of computed tomography in detecting asbestos-related pleural abnormalities in people who had indeterminate chest radiographs: the Libby, MT, experience. AB - This epidemiological study was conducted to determine whether high-resolution computed tomography (HRCT) is useful to screen for pulmonary abnormalities in people exposed to vermiculite containing asbestos. During June-September 2001, we evaluated HRCT of 353 people in Libby, MT, who had been exposed to asbestiform minerals associated with vermiculite. Of these, 334 participants of the summer 2000 medical testing program underwent HRCT of the chest at St. John's Lutheran Hospital and 19 eligible people who recently had undergone an HRCT scan at the same facility and under the same testing protocol allowed the study reviewers to use that scan. All 353 study participants were former vermiculite mine/mill workers (n = 55), their household contacts (n = 99), and people exposed to vermiculite through recreational or other activities (n = 199). Participants' 2000 medical testing results indicated only one of the three B-reader chest radiograph reviewers had reported a pleural abnormality (indeterminate chest radiograph). Three expert computer tomography (CT) scan evaluators reviewed the HRCT scans and identified pleural abnormalities in 98 (27.8%) of the 353 participants whose previous chest radiographs were classified indeterminate. Of these 98 people, 69 (70.4%) were either former vermiculite mine/mill workers or household contacts, and 40 (40.8%) showed pleural calcification on HRCT. Thirty out of the 40 people with pleural calcification reported having no occupational exposure to either Libby vermiculite or asbestos. Our findings indicate that low dose HRCT can be considered for screening certain former vermiculite mine/mill workers and their household contacts who have indeterminate chest radiographs and may be useful for diagnosing a suspicious finding on a chest radiograph, particularly in a high-risk person. PMID- 15881983 TI - Community environmental health assessment strengthens environmental public health services in the Peruvian Amazon. AB - In December 1999, the Centers for Disease Control and Prevention (CDC) and the Cooperative for Assistance and Relief Everywhere, Peru Country Office (CARE Peru), initiated the Urban Environmental Health Project (SAU, in Spanish) to strengthen environmental public health services in urban and periurban settlements in Peru. The project received funding from the Woodruff Foundation as part of the CARE-CDC Health Initiative (CCHI). The "Protocol for Assessing Community Excellence in Environmental Health" (PACE EH) guided the development of a community environmental public health assessment (CEHA) process in Cardozo, a settlement in Iquitos, Peru. The project developed a three-phase process that merged scientific understanding and community perception about local environmental health problems. In phase 1, local environmental health technicians assisted the community in understanding environmental health conditions in Cardozo and selecting priorities. During phase 2, local technicians assessed the community-selected priorities: water and sanitation. Results from recent water quality assessments revealed that 82% (9 of 11) of samples from shallow dug wells, 18% (2 of 11) from deeper drilled wells, and 61% (11/18) from household drinking containers were positive for thermotolerant coliforms. Phase 3 activities produced an action plan and an intervention to mitigate health problems associated with inadequate water and sanitation services in the Cardozo community. As a result of the CEHA process, CARE Peru obtained funding from the United States Agency for International Development (USAID) to develop and implement an environmental health risk monitoring system and the proposed water and sewage intervention in the settlement. CDC continues to provide technical assistance to the local environmental health services groups in Iquitos through an agreement with CARE Peru as part of the USAID-funded Urban Environmental Health Models Project (MUSA). Technical assistance activities and the development of the environmental health risk monitoring system have helped to strengthen the local environmental public health services delivery system. PMID- 15881984 TI - The utility of geographic information systems (GIS) in rapid epidemiological assessments following weather-related disasters: methodological issues based on the Tropical Storm Allison Experience. AB - Flooding is the most common natural disaster worldwide, and is the leading cause of weather-related deaths in the United States. Tropical storm Allison hit landfall near Galveston, Texas on June 5, 2001, causing the most severe flood related damage ever recorded in the Houston metropolitan area. This devastating storm dumped 37 in of rain in 24h on parts of the city, killing 22 people and causing more than $5 billion in damage. The main goal of the public health response to tropical storm Allison was to rapidly evaluate the immediate health needs of the community. Geographical information system (GIS) technology was instrumental to the timeliness of this effort. We conducted a rapid needs assessment in the areas most affected by flooding using modified cluster sampling facilitated by GIS methodology. Of the 420 households participating in the survey, we found a significant increase in illness (OR, 5.1; 95% CI, 2.7-9.4), injuries (OR, 4.8; 95% CI, 1.9-12.8), and immediate health needs (OR, 3.3; 95% CI, 1.7-6.1) among persons living in flooded homes compared to non-flooded homes. There were 60 households reporting serious damage, 24 of which were outside the 500-year flood plain. We also obtained reliable estimates of the extent of damage and household needs to help guide relief efforts. These findings underscore the usefulness of rapid needs assessment as a tool to identify actual health threats and to facilitate delivery of resources to those with the greatest and most immediate need. Our ability to swiftly plan and implement a rapid needs assessment over a large geographical region within 1 week following the damage would not have been possible without the utilization of GIS methodology and the availability of skilled personnel and timely data resources. PMID- 15881985 TI - Systems modeling in support of evidence-based disaster planning for rural areas. AB - The objective of this communication is to introduce a conceptual framework for a study that applies a rigorous systems approach to rural disaster preparedness and planning. System Dynamics is a well-established computer-based simulation modeling methodology for analyzing complex social systems that are difficult to change and predict. This approach has been applied for decades to a wide variety of issues of healthcare and other types of service capacity and delivery, and more recently, to some issues of disaster planning and mitigation. The study will use the System Dynamics approach to create computer simulation models as "what if" tools for disaster preparedness planners. We have recently applied the approach to the issue of hospital surge capacity, and have reached some preliminary conclusions--for example, on the question of where in the hospital to place supplementary nursing staff during a severe infectious disease outbreak- some of which we had not expected. Other hospital disaster preparedness issues well suited to System Dynamics analysis include sustaining employee competence and reducing turnover, coordination of medical care and public health resources, and hospital coordination with the wider community to address mass casualties. The approach may also be applied to preparedness issues for agencies other than hospitals, and could help to improve the interactions among all agencies represented in a community's local emergency planning committee. The simulation models will support an evidence-based approach to rural disaster planning, helping to tie empirical data to decision-making. Disaster planners will be able to simulate a wide variety of scenarios, learn responses to each and develop principles or best practices that apply to a broad spectrum of disaster scenarios. These skills and insights would improve public health practice and be of particular use in the promotion of injury and disease prevention programs and practices. PMID- 15881986 TI - Occupational physician perceptions of bioterrorism. AB - The rationale for most preparedness training of healthcare professionals is based on the assumption that most persons infected following a biological incident will present first to emergency departments of acute care facilities or to ambulatory settings such as private physician offices, and such incidences would be recognized, appropriately treated, and reported to the local health departments. However, an alternative first point of contact is industry, a location where workers gather and disperse on a regular and documented basis, and require healthcare. In industry there are health professionals responsible for the health, safety and on-site well-being of the workforce and surrounding community; these professionals are in a position for early recognition, surveillance, and isolation. Targeted education must be provided to these health professionals. To address perceptions of risk and preferred educational delivery methods for bioterrorism and emerging infections-related materials, a survey of occupational physicians was performed during the spring of 2001. Within the 2 months following the September 11 terrorist attack and subsequent anthrax bioterrorism event, and before release of any results from the first survey, a follow-up mail survey was initiated in November 2001. Response rate to the pre- and post-September 11 survey were 58% (n = 56) and 33% (n = 33), respectively. No significant demographic differences were observed between the respondents of the pre- and post-surveys. Perceptions of likelihood of another bioterrorism event increased between surveys, as would be expected; however, a tendency to believe that it would not happen locally persisted. Even though over 90% of the physicians had received immediate training following September 11, additional training/education needs were demonstrated. Although training and education modules can be designed without information based on the population that can be on the receiving end, it rarely accomplishes its goal. Results from this survey can serve as a base for designing various levels of targeted training and educational material specific to the perceived need, method of obtaining information and the format considered to be most conducive for learning. Potential consequences from lack of bioterrorism preparedness due to low perception of need and threat awareness need to be addressed. PMID- 15881987 TI - Perspectives on the use of scientific knowledge for public health practice. AB - Achieving the goal of increasing quality and years of healthy life is fundamentally based on success in the practice of public health. As our life style changes with time and as public health issues become more global, the practice of public health is enhanced to meet new challenges. In addition to addressing infectious diseases, environmental concerns are gaining attention. New challenges require the modification of the methods of investigations, use of new technologies and application of real-time management of public health emergencies. In many situations, collaborations at the local, regional, national and global levels are needed. This manuscript provides a summary of the approaches to address certain crucial environmental health concerns towards the goal of increasing quality and years of healthy life. PMID- 15881988 TI - The cutting edge. Veterinarians discuss applying science to practice. PMID- 15881989 TI - PRRSV planned exposure and the law. PMID- 15881990 TI - Carlson seeking second term as AVMA vice president. Interview by Susan C. Kahler. PMID- 15881991 TI - Questions findings in Spanish Mustang study. PMID- 15881992 TI - Is there a place for lawyers in animal rights issues? PMID- 15881993 TI - Is there a place for lawyers in animal rights issues? PMID- 15881994 TI - Distinction between Hepatozoon canis and Hepatozoon americanum. PMID- 15881995 TI - Looking deeper into quality-of-life assessment. PMID- 15881996 TI - The allure and the illusion of science. PMID- 15881997 TI - What is your diagnosis? Metal foreign body reaction complicated by a staphylococcal infection. PMID- 15881998 TI - ECG of the month. Accelerated idioventricular rhythm. PMID- 15881999 TI - Growth hormone-responsive alopecia in dogs. PMID- 15882000 TI - Accuracy of polymerase chain reaction assays for diagnosis of feline immunodeficiency virus infection in cats. AB - OBJECTIVE: To determine the sensitivity, specificity, and overall diagnostic accuracy of polymerase chain reaction (PCR) assays offered by commercial diagnostic laboratories for diagnosis of FIV infection in cats. DESIGN: Prospective clinical trial. ANIMALS: 124 cats. PROCEDURE: Blood was collected from cats that were neither infected with nor vaccinated against FIV, uninfected cats that were vaccinated with a licensed FIV vaccine, and cats experimentally and naturally infected with FIV representing subtypes A, B, and C. Coded blood samples were submitted to 3 laboratories in the United States and Canada offering PCR assays for diagnosis of FIV infection to veterinary practitioners. All laboratories tested fresh blood samples, and 1 laboratory also tested samples submitted as dried blood smears. The FIV infection status in all cats was confirmed by virus isolation. Sensitivity, specificity, and correct results were calculated for each PCR assay. RESULTS: Sensitivity ranged from 41% to 93%. Specificity ranged from 81% to 100% in unvaccinated cats and 44% to 95% in cats vaccinated against FIV. Correct results were obtained in 58% to 90% of 124 cats tested. All tests misidentified both uninfected and infected cats. False-positive results by all laboratories were higher in cats vaccinated against FIV than in unvaccinated cats, suggesting that vaccination interferes with the performance or interpretation of PCR assays used for diagnosis of FIV infection. CONCLUSIONS AND CLINICAL RELEVANCE: PCR assays used for diagnosis of FIV infection presently marketed to veterinary practitioners in North America vary significantly in diagnostic accuracy and did not resolve the diagnostic dilemma resulting from vaccination of cats against FIV. PMID- 15882001 TI - Technique and outcome of mitral valve replacement in dogs. AB - Eight dogs with naturally occurring severe mitral regurgitation underwent mitral valve replacement with a mechanical valve prosthesis during cardiopulmonary bypass. Dogs received warfarin orally after surgery to maintain a prothrombin time-based international normalized ratio from 2.5 to 3.5. Seven dogs survived surgery. Left ventricular diastolic volume index decreased significantly from 206 +/- 91 mL/m2 before surgery to 121 +/- 47 mL/m2 after surgery. Left atrium-to aorta ratio decreased significantly from 2.66 +/- 0.4 before surgery to 1.73 +/- 0.65 after surgery. Left ventricular systolic volume index was not significantly different after surgery (56 +/- 36 mL/m2), compared with before surgery (40 +/- 32 mL/m2). Median survival after surgery was 4.5 months (range, 0.75 months to 5.25 years). Six dogs died of confirmed or suspected thrombosis of the valve prosthesis. Dogs with severe mitral regurgitation tolerated mitral valve replacement well, but a high incidence of prosthetic valve thrombosis limited long-term outcome. PMID- 15882002 TI - Aneurysmal dilatation of the right auricle in two dogs. AB - An 8-year-old sexually intact male Golden Retriever with a history of collapse during exercise underwent an examination during which tachydysrhythmia was identified. At another institution, a 12.5-year-old spayed female Lhasa Apso was referred because of a cough and for evaluation of a heart murmur. In the Golden Retriever, radiographic examination revealed bulging of the craniodorsal aspect of the cardiac silhouette and echocardiography revealed right atrial dilatation. In the Lhasa Apso, a cranial mediastinal mass was suspected on the basis of radiographic findings, but no abnormalities were detected echocardiographically. In both dogs, nonselective angiography and contrast-enhanced computed tomography revealed a markedly enlarged, thin-walled right auricle. Exploratory thoracotomy in the Golden Retriever revealed a defect in the pericardium through which the right auricle and part of the atrium had herniated. In dogs, a right auricular aneurysm should be considered in differential diagnoses of a cranial mediastinal mass (detected radiographically) adjacent to the cardiac silhouette. PMID- 15882003 TI - Heinz body anemia in a dog that had been sprayed with skunk musk. AB - A 5-year-old pit bull-type dog was examined because of lethargy and black feces, and a presumptive diagnosis of methemoglobinemia was made on the basis of grossly visible brown discoloration of the mucous membranes and urine. Heinz body formation was seen on examination of blood smears, and severe anemia developed 3 days later. The dog did not have any history of exposure to toxins or drugs that have been associated with Heinz body formation. The only remarkable historical finding was that the dog had been wandering loose the day prior to initial examination and returned home smelling strongly of skunk spray. Skunk spray contains thiols and other compounds that are strong oxidizing agents. It was speculated that the methemoglobinemia and Heinz body anemia were a result of exposure to these compounds. Skunk interactions with dogs usually have malodorous but otherwise harmless results. However, findings in this dog suggest that more serious consequences may develop in isolated cases. PMID- 15882004 TI - Dysphagia as the primary clinical abnormality in two dogs with inflammatory myopathy. AB - Two adult Boxers were evaluated because of chronic dysphagia of several years' duration. Serum creatine kinase activity was high in both dogs, but other hematologic or serum biochemical abnormalities were not detected. Esophagraphy revealed abnormalities of the cricopharyngeal phase of swallowing in both dogs, and electromyography of the pharyngeal and laryngeal muscles revealed complex repetitive discharges, positive sharp waves, and fibrillation potentials characteristic of primary myopathy or neuropathy. Because of the severity of their condition, both dogs were euthanatized. Histologically, mixed-cell infiltrates were seen in sections of the masseter and thyropharyngeal muscles. Results of indirect immunofluorescence staining for proteins associated with dystrophic myopathy were unremarkable, except for decreased staining for integrin alpha7. A diagnosis of chronic inflammatory myopathy was made. The clinical importance of reduced staining for integrin alpha7 could not be determined but was considered to be a result of the myopathy. PMID- 15882005 TI - Comparison of two techniques for injection of the podotrochlear bursa in horses. AB - OBJECTIVE: To compare accuracy of 2 methods for injection of the podotrochlear bursa in horses. DESIGN: Observational study. ANIMALS: 17 French Standardbreds. PROCEDURE: In each horse, contrast medium was injected into the podotrochlear bursa of 1 foot by use of a distal palmar approach with the needle inserted parallel to the sole (DPPS) and into the podotrochlear bursa of the other foot by use of a distal palmar approach to the navicular position (DPNP). Podotrochlear bursa injection was evaluated by means of radiographic examination in all horses and postmortem examination in 6. RESULTS: Contrast medium was successfully injected into the podotrochlear bursa in 6 of 16 feet in which the DPPS method was used and 14 of 17 feet in which the DPNP method was used; these results were significantly different. Failure was attributed to contrast medium invading the distal interphalangeal joint, contrast medium pooling palmar to the deep digital flexor tendon, contrast medium located in the podotrochlear bursa and the distal interphalangeal joint, contrast medium located in the podotrochlear bursa and the area palmar to the deep digital flexor tendon, and an inability to inject contrast medium despite adequate needle placement. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the DPNP technique can be used successfully for injection into the podotrochlear bursa in horses. However, radiographic assessment should be used to confirm that material was injected into the bursa. PMID- 15882006 TI - Prevalence and risk factors associated with outcome of surgical removal of pedunculated lipomas in horses: 102 cases (1987-2002). AB - OBJECTIVE: To determine the prevalence of pedunculated lipomas and identify risk factors affecting postoperative complications and survival in horses at a veterinary teaching hospital undergoing surgery for colic caused by pedunculated lipomas. DESIGN: Retrospective study. ANIMALS: 102 horses with a diagnosis of pedunculated lipoma. PROCEDURE: Age, breed, weight, and sex of horses with pedunculated lipomas were compared with the total equine hospital population and the population of horses admitted for abdominal surgery during the same period. Follow-up information was obtained by reevaluation or contact with owners via telephone or written request. RESULTS: Prevalence of pedunculated lipomas as a reason for abdominal surgery in horses, compared with the population of horses with and without lipomas admitted for abdominal surgery, was 10%. Castrated male Saddlebred and Arabian horses > 14 years old were identified as being at risk for developing pedunculated lipomas. Postoperative complications were detected in 72% of horses with pedunculated lipomas. Variables associated with low survival rates included surgery before 1992, heart rate > 80 beats/min, abnormal color of abdominal fluid, pale mucous membranes, surgery requiring intestinal resection, and inability to attain a mean arterial pressure > or = 100 mm Hg. Horses undergoing surgery from 1992 to 1996, weighing < 409 kg (900 lb), or requiring jejunojejunal anastomosis had a high survival rate. CONCLUSIONS AND CLINICAL RELEVANCE: Although many of the variables reflected the health of the horse at the time of surgery, results may help veterinarians recognize risk factors associated with development of pedunculated lipomas and better predict the outcome of horses undergoing surgery for colic caused by pedunculated lipomas. PMID- 15882007 TI - Results of a national survey of US veterinary college faculty regarding attitudes toward farm animal welfare. AB - OBJECTIVE: To examine attitudes toward farm animal welfare among veterinary college faculty. DESIGN: E-mail survey. STUDY POPULATION: 157 US veterinary college faculty with large animal or food animal emphasis. PROCEDURE: Veterinarians from 27 US veterinary colleges were contacted via e-mail and asked to complete a 7-page survey relating to farm animal welfare issues. Thirty-one percent of those contacted responded. RESULTS: 71% of respondents self characterized their attitude toward farm animal welfare as "we can use animals for the greater human good but have an obligation to provide for the majority of the animals' physiologic and behavioral needs." An additional 19% of respondents were more concerned about animal welfare than was indicated by that statement, and 10% were less concerned about farm animal welfare than was indicated by that statement. Significant relationships among demographic variables and attitude scores were observed, including more concerned attitudes among females, those with more liberal political views, and those who cited lower religiosity. No relationship between attitude and age was observed. CONCLUSIONS AND CLINICAL RELEVANCE: Veterinary college faculty have the opportunity to impact many stakeholders within the animal agriculture industries (eg, future veterinarians and policy makers looking for a veterinary science perspective). Results indicated that a considerable level of concern toward farm animal welfare is present in this population. Although the process of change may not be rapid, it is likely that the influence of these respondents will factor heavily into enhancing farm animal welfare. PMID- 15882008 TI - Economic analysis of feeding pasteurized nonsaleable milk versus conventional milk replacer to dairy calves. AB - OBJECTIVE: To determine growth, morbidity, and mortality rates in dairy calves fed pasteurized nonsaleable milk versus commercial milk replacer and compare economics of feeding pasteurized nonsaleable milk versus commercial milk replacer in dairy calves. DESIGN: Clinical trial. ANIMALS: 438 dairy calves. PROCEDURE: Calves were assigned at 1 to 2 days of age to be fed pasteurized nonsaleable milk or a commercial milk replacer until weaned. Body weight was measured at the time of study enrollment and at the time of weaning, and any medical treatments administered and deaths that occurred prior to weaning were recorded. A partial budget model was developed to examine the economics of feeding pasteurized nonsaleable milk versus commercial milk replacer. RESULTS: Calves fed conventional milk replacer had significantly lower rates of gain (-0.12 kg/d [ 0.26 lb/d]), lower weaning weights (-5.6 kg [-12.3 lb]), higher risk for treatment during the summer and winter months (odds ratio [OR], 3.99), and higher risk of death during the winter months (OR, 29.81) than did calves fed pasteurized nonsaleable milk. The estimated savings of feeding pasteurized nonsaleable milk, compared with milk replacer, was dollars 0.69/calf per day. The estimated number of calves needed to economically justify the nonsaleable milk pasteurization system was 23 calves/d. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dairy calves fed pasteurized nonsaleable milk have a higher growth rate and lower morbidity and mortality rates than do calves fed conventional milk replacer. Feeding pasteurized nonsaleable milk could be an economically viable strategy for dairy calf producers. PMID- 15882009 TI - Idiopathic pericarditis and cardiac tamponade in two cows. AB - Idiopathic pericarditis is an uncommon diagnosis in cattle with cardiac tamponade. Two cows were examined for clinical signs of right-sided congestive heart failure, including tachycardia, venous distention, and peripheral edema. Muffled heart sounds were detected in one of the cows. Echocardiography in both cows revealed voluminous anechoic pericardial effusion and compression of the right atrium and right ventricle. Cytologic analysis of the pericardial fluid revealed hemorrhagic inflammation but no evidence of a septic or neoplastic condition such as traumatic reticulopericarditis or lymphoma, respectively. Pericardial drainage and lavage accompanied by treatment with anti-inflammatory drugs were curative in both cows. It is important to differentiate cows with idiopathic pericarditis from cows with more common septic pericarditis because the prognosis for the former disease appears to be good with appropriate treatment. PMID- 15882010 TI - Human inventory control. PMID- 15882012 TI - Too cold for comfort. PMID- 15882011 TI - Cooping up avian flu. PMID- 15882013 TI - Defensive eating. PMID- 15882014 TI - Bending to bar codes. PMID- 15882015 TI - Doubts on dinosaurs. PMID- 15882016 TI - Processing for science. PMID- 15882017 TI - Measuring beauty. PMID- 15882018 TI - Turn me on, dead man. PMID- 15882019 TI - When medicine meets literature. PMID- 15882021 TI - Quantum black holes. PMID- 15882020 TI - His brain, her brain. AB - It turns out that male and female brains differ quite a bit in architecture and activity. Research into these variations could lead to sex-specific treatments for disorders such as depression and schizophrenia PMID- 15882022 TI - Neuromorphic Microchips. AB - Compact, efficient electronics based on the brain's neural system could yield implantable silicon retinas to restore vision, as well as robotic eyes and other smart sensors. PMID- 15882023 TI - A bolt out of the blue. AB - Lightning is a particularly unsettling product of bad weather. It causes more deaths and injuries in the U.S. than either hurricanes or tornadoes do, and it strikes without warning, sometimes with nothing but blue sky overhead. In central Florida, where I live, thunderstorms are a daily occurrence during the summer, and so, ironically, people in the Sunshine State often spend their afternoons indoors to avoid the risk of death from the sky. Worldwide, lightning flashes about four million times a day, and bolts have even been observed on other planets. Yet despite its familiarity, we still do not know what causes lightning. It is a misconception that Benjamin Franklin solved the puzzle when he conducted his famous kite experiment in 1752. PMID- 15882024 TI - Can chlamydia be stopped? AB - Chlamydia is a rampant sexually transmitted disease, the world's leading cause of preventable blindness and a possible contributor to heart disease. Recent discoveries are suggesting new ways to curtail its spread PMID- 15882025 TI - What heated the asteroids? PMID- 15882026 TI - Molecular treasure hunt. PMID- 15882027 TI - [Chronic irradiation with low doses can be characterized by significant biological efficacy]. AB - Chromosomal aberrations (ChA) level was analyzed in the onion root meristem after the chronic irradiation with different dose capacities. It was shown that after the chronic irradiation with doses of 0.87 cGy, 2.61 cGy and 4.35 cGy the level of chromosomal aberrations depended on the dose capacity. Its value may also correspond to those which have been induced with accute irradiation. Biological efficacy of chronic irradiation may be from 20 to 1000 time folder in order to compare it with accute irradiation and this value depends on the irradiation regime. PMID- 15882028 TI - [Circadian mitotic rhythm in representatives of the soybean Glycine L]. AB - The circadian mitotic rhythms in representatives of soybean Glycine max (L.) Merr., G. soja Sieb. & Zucc., as well as in homozygote forms of tibia F6 hybrids were studied. We observed three "waves" of the mitotic activity in the cells of apical root meristem with the initial moment of the perfect rest at 24 o'clock; and the legible periods which decreased later on. Beginning from 1 o'clock to 6 o'clock the first maximum mitotic "wave" increased; the second one evolved up to 12 o'clock, the third less legible one arised at 18-21 o'clock. Each of soybean tested forms was distinguished by the peculiarity in periodical processes and the values of mitotic indexes PMID- 15882029 TI - [Mutation variability and marker trait inheritance in hybrid-mutant populations F2M2 of winter wheat]. AB - Influence of hybrid seed treatment with mutagenes on genetic variability in hybrid populations of the second generation of winter wheat has been investigated. Treatment with mutagenes in moderate doses results in deviation of segregation of marker traits as compared to control towards either increase or decrease of the ratio of recessive allels. It was shown that in crossing combinations with red-ear form as one of the parental components, treatment with gamma rays at the dose of 100 Gy leads to significantly larger deviations of disintegration comparing to control. PMID- 15882030 TI - [Reconstruction of the origin of modern swine breeds by mitochondrial gene polymorphism]. AB - Mitochondrial DNAs of six pig breeds of Euro-American origin, one breed of Asian origin and of wild boar were studied using PCR-RFLP analysis to detect probable maternal lines of Italian origin among modern pig breeds. In the studied animals the Italian Wild boar haplotype characterized by single nucleotide substitution in the 15524 position of pig mitohondrial genome has not been detected. PMID- 15882031 TI - [Ultrastructural changes in loach (Misgurnus fossilis L.) embryos under borocin treatment]. AB - The ultrastructure of embryo cells of the loach (Misgurnus fossilis L.) at the stage of first division of blastomers in control and under the conditions of fluoroquinole borocin treatment has been investigated. The influence of this antibiotic at concentrations 5 and 25 mkg/ml has resulted in significant ultrastructural changes of embryo cells, such as hypertrophy of channels of the smoth and rough endoplasmic reticulum, disorganization of Golgy complex and mitochondrias, destruction of cytoplasm and mitochondrial membranes, rarefaction of cytoplasm and cell oedema. Such changes confirm the toxic influence of borocin on the embryo during early development. PMID- 15882032 TI - [Meiotic chromosomes of green frogs (Rana esculenta complex) of the Ukraine territory]. AB - In this work meiotic chromosomes of R. ridibunda, R. lessonae and their hybrid form R. esculenta of the Ukraine territory are described for the first time. These chromosomes are connected in diakinesis more often by two chiasma, rarely by one or three chiasma in 13 bivalents. Bivalents with one chiasma in R. ridibunda are typical only for large bivalents, in R. lessonae only for small ones. Three chiasma in parental species are typical only for small bivalents. In the hybrid form R. esculenta chromosomes with two, one and three chiasma are typical both for large bivalents, and for small ones. In the hybrid form (R. esculenta) chiasma frequency noticeably decreases, comparatively with parental species that, possibly, specifies the aberration of normal meiosis realization. The chiasma frequencies of the hybrid form of the eastern and the southern parts of Ukraine are almost the same, but the types of chromosomes are different in such regions. In the eastern region chromosomes of the hybrid form are represented by two or one chiasma, in the southern region by two, one or three chiasma. PMID- 15882033 TI - [Role of non-radiation factors in forming cytogenetic effects in evacuees from a 30-km zone of the Chernobyl Nuclear Power Plant]. AB - The time-effect relationship for chromatid type aberrations, chromosome type fragments, hyperploidy and polyploidy levels in peripheral blood lymphocytes were investigated in inhabitants of t. Pripiat' and nearby villages, who were departured from the Chernobyl NPP 30-km exclusive zone during first days after the Chernobyl catastrophe. The time-course changes of the mentioned cytogenetic indices in evacuees were displayed as a gradual decline of chromosomal rearrangements and genome abnormality frequencies from the statistically elevated level in the first 1-2 years after the accident to the subcontrol meanings at the end of the 14-years observation period. In early terms after exposure the frequency of chromatid exchanges in adult men and the polyploidy level in women aged 23-35 years were sufficiently increased comparing with other evacuee subgroups. Some peculiarities of the fragment aberration frequency dynamics were shown for persons with different terms of the departure from the Chernobyl zone. The role of the combination of mutagenic factors acted in the accidental situation at Chernobyl for inducing the elevated level of cytogenetic damage in evacuees is discussed. PMID- 15882034 TI - [The level of spontaneous chromosome aberrations in children from ecologically clean region of Ukraine studied with cytogenetic analysis of uniformly and differentially stained metaphase chromosomes]. AB - The frequency of chromosome aberrations in children from Lviv region of Ukraine born after Chernobyl accident have been determined using conventional and G banding staining. PMID- 15882035 TI - [Estimation of potential mutagenic activity of 24-epibrassinolide]. AB - Genotoxic effect of synthetic phytohormone analogue of steroid origin epibrassinolide was studied in in vitro- and in vivo-tests. Epibrassinolide did not display mutagenic properties in Ames' test (S. typhimurium, TA100) and DNA damaging activity test (DNA of phage lambda). The rise in the level of polichromatophilous erythrocytes with micronuclei was observed in the micronuclear test at intraperitoneal epibrassinolide injection at the dose of 500 mg/kg that seems to be associated with disturbance of cell membrane permeability. PMID- 15882036 TI - [Molecular cytogenetic study of Robertsonian translocation 13;14 and Down syndrome in a 3-year-old infant]. AB - We describe here a rare case of Robertsonian translocation 13;14 of maternal origin combined with regular trisomy 21 (46,XX,der(13;14)(q10;q10)mat,+21) with Down syndrome phenotype. Molecular cytogenetic studies allowed us to determine the maternal origin of additional chromosome 21 and the non-disjunction of chromosome 21 to occur in meiosis I. On the basis of data obtained we discuss the possible involvement of structural alterations of chromosomes 13 and 14 in the chromosome 21 non-disjunction. PMID- 15882037 TI - [Cytomorphological characteristics of normal lymphatic gland cell structure]. AB - Numerical and qualitative characteristics of cell structure of 87 normal human lymphatic gland cytograms of various age groups have been studied. For each age group distinctive cell profiles of cytograms were established and cytological features of their development and involution have been determined. PMID- 15882038 TI - [Study of the repair enzyme O6-alkylguanine-DNA-alkyltransferase presence in mammalian cells in vitro]. AB - The enzyme O6-alkylguanine-DNA-alkyltransferase (AGT) has been revealed to be present in human cell cultures (fibroblasts and cardiomyocytes) and Chinese hamster culture cells. At the same time the enzyme AGT is not reproduced in the human brain shell culture cells. PMID- 15882040 TI - Spinal cord injury and obesity. PMID- 15882039 TI - [Prevention of inborn hereditary defects and inherited pathology (from material of a conference)]. PMID- 15882041 TI - Canadian pilot study eliminates wait times for dialysis patients. PMID- 15882042 TI - What's in a number? PMID- 15882043 TI - Impact of multidisciplinary, early renal education on vascular access placement. AB - BACKGROUND: The Ochsner Clinic Foundation initiated the Healthy Start Clinic to identify, educate, and refer chronic kidney disease patients to nephrologists earlier in the course of their disease. This study investigated the impact of a structured educational session on the type and timing of permanent vascular access placement in patients receiving hemodialysis. METHODS: Before initiating dialysis, the HSC patient group received a general overview of the kidney and kidney disease, plus one-on-one instruction from a registered nurse, a dietitian, and a social worker. The HSC group was compared with a concurrent, conventionally prepared group of CKD patients who initiated dialysis during the same study period. RESULTS: Of the 147 patients initiating hemodialysis at OCF between April 1997 and December 2000, 61 had received structured HSC education, and 86 had received conventional care. In HSC-educated patients, the incidence of PVAs placed before hemodialysis initiation was twofold greater than in patients who received conventional care (77% HSC; 36% non-HSC, p < 0.001), and more HSC educated patients initiated hemodialysis using their PVA (49% HSC; 23% non-HSC, p < 0.01). In addition, five times more patients who received HSC education received arteriovenous fistulas (52% HSC; 10% non-HSC, p < 0.001). Finally, significantly fewer HSC-educated patients initiated dialysis on a temporary catheter (51% HSC; 77% non-HSC, p < 0.001). CONCLUSIONS: Education programs for CKD patients help increase the number of patients receiving early PVA placement, as well as the proportion of patients receiving AVFs as opposed to grafts or temporary catheters. PMID- 15882044 TI - Overcoming the ouch: addressing patient needle fears in the dialysis clinic. PMID- 15882045 TI - The bare essentials: dialyzing in Russia. PMID- 15882046 TI - Finding dialysis care in Mexico: neither fiesta nor siesta. PMID- 15882047 TI - Providing illegal immigrants with dialysis: the Maryland experience. PMID- 15882048 TI - Analysis of the 2.0 A crystal structure of the protein-DNA complex of the human PDEF Ets domain bound to the prostate specific antigen regulatory site. AB - PDEF, a prostate epithelial specific transcription factor, is a member of the Ets family of DNA binding proteins. Here we report a 2.0 A crystal structure of the PDEF Ets domain in complex with a natural, high-affinity DNA binding site in the promoter/enhancer region of the human prostate specific antigen gene. Comparison of the PDEF-DNA complex with other Ets complexes revealed key features that are shared among Ets members, as well as important differences in substrate specification at both the "GGA" core and the flanking regions of the DNA site. The combination of the serine residue at position 308 and the glutamine at position 311 explains the previous observation that the PDEF binds preferentially to a thymine at the +4 position of its binding site. Despite the common essential features that are shared among Ets members, PDEF demonstrates distinct patterns of interactions at different positions of DNA in achieving sequence specific recognition. Collectively, the common and unique interactions with both the DNA bases and the backbone phosphates lead to substrate specificity and individual preference for certain DNA sites. PMID- 15882049 TI - A protecting group for carboxylic acids that can be photolyzed by visible light. AB - We report on a photolabile protecting (caging) group that is new for carboxylic acids. Unlike previously used caging groups for carboxylic acids, it can be photolyzed rapidly and efficiently in the visible wavelength region. The caging group 7-N,N-diethyl aminocoumarin (DECM) was used to cage the gamma-carboxyl group of glutamic acid, which is also a neurotransmitter. The caged compound has a major absorption band with a maximum at 390 nm (epsilon(390) = 13651 M(-)(1) cm(-)(1)). Experiments are performed at 400 nm (epsilon(400) = 12232 M(-)(1) cm( )(1)) and longer wavelengths. DECM-caged glutamate is water soluble and stable at pH 7.4 and 22 degrees C. It photolyzes rapidly in aqueous solution to release glutamic acid within 3 micros with a quantum yield of 0.11 +/- 0.008 in the visible region. In whole-cell current-recording experiments, using HEK-293 cells expressing glutamate receptors and visible light for photolysis, DECM-caged glutamate and its photolytic byproducts were found to be biologically inert. Neurotransmitter receptors that are activated by various carboxyl-group containing compounds play a central role in signal transmission between approximately 10(12) neurons of the nervous system. Caged neurotransmitters have become an essential tool in transient kinetic investigations of the mechanism of action of neurotransmitter receptors. Previously uncaging the compounds suitable for transient kinetic investigations required ultraviolet light and expensive lasers, and, therefore, special precautions. The availability of caged neurotransmitters suitable for transient kinetic investigations that can be photolyzed by visible light allows the use of simple-to-use, readily available inexpensive light sources, thereby opening up this important field to an increasing number of investigators. PMID- 15882050 TI - Mechanism of the reaction catalyzed by isoaspartyl dipeptidase from Escherichia coli. AB - Isoaspartyl dipeptidase (IAD) is a member of the amidohydrolase superfamily and catalyzes the hydrolytic cleavage of beta-aspartyl dipeptides. Structural studies of the wild-type enzyme have demonstrated that the active site consists of a binuclear metal center positioned at the C-terminal end of a (beta/alpha)(8) barrel domain. Steady-state kinetic parameters for the hydrolysis of beta aspartyl dipeptides were obtained at pH 8.1. The pH-rate profiles for the hydrolysis of beta-Asp-Leu were obtained for the Zn/Zn-, Co/Co-, Ni/Ni-, and Cd/Cd-substituted forms of IAD. Bell-shaped profiles were observed for k(cat) and k(cat)/K(m) as a function of pH for all four metal-substituted forms. The pK(a) of the group that must be unprotonated for catalytic activity varied according to the specific metal ion bound in the active site, whereas the pK(a) of the group that must be protonated for catalytic activity was relatively independent of the specific metal ion present. The identity of the group that must be unprotonated for catalytic activity was consistent with the hydroxide that bridges the two divalent cations of the binuclear metal center. The identity of the group that must be protonated for activity was consistent with the free alpha-amino group of the dipeptide substrate. Kinetic constants were obtained for the mutant enzymes at conserved residues Glu77, Tyr137, Arg169, Arg233, Asp285, and Ser289. The catalytic properties of the wild-type and mutant enzymes, coupled with the X-ray crystal structure of the D285N mutant complexed with beta-Asp-His, are consistent with a chemical reaction mechanism for the hydrolysis of dipeptides that is initiated by the polarization of the amide bond via complexation to the beta metal ion of the binuclear metal center. Nucleophilic attack by the bridging hydroxide is facilitated by abstraction of its proton by the side chain carboxylate of Asp285. Collapse of the tetrahedral intermediate and cleavage of the carbon-nitrogen bond occur with donation of a proton from the protonated form of Asp285. PMID- 15882051 TI - DNA-based biosensor for monitoring pH in vitro and in living cells. AB - DNA is a promising material for the construction of a biosensor or bioindicator because its structure is sensitive to the binding of cofactors. In the current studies, we found that a combination of two DNA oligonucleotides, 5'-TCTTTCTCTTCT 3' and 5'-AGAAAGAGAAGA-3', exhibit a novel structural transition from a Watson Crick antiparallel duplex to a parallel Hoogsteen duplex as the pH changes from pH 7.0 to 5.0. By labeling this DNA for fluorescence resonance energy transfer, we were able to develop a sensitive pH indicator that can detect changes between pH 7.0 and 5.0. Moreover, using DNA-based hairpin parallel-stranded duplex in conjunction with fluorescence microscopy, we were able to observe the pH changes in living cells during apoptosis as an easily detected change in color. These results indicate that the DNA-based pH indicator should be useful for detecting pH changes between pH 7.0 and 5.0 in living cells. PMID- 15882052 TI - Crystal structures and functional studies of T4moD, the toluene 4-monooxygenase catalytic effector protein. AB - Toluene 4-monooxygenase (T4MO) is a four-component complex that catalyzes the regiospecific, NADH-dependent hydroxylation of toluene to yield p-cresol. The catalytic effector (T4moD) of this complex is a 102-residue protein devoid of metals or organic cofactors. It forms a complex with the diiron hydroxylase component (T4moH) that influences both the kinetics and regiospecificity of catalysis. Here, we report crystal structures for native T4moD and two engineered variants with either four (DeltaN4-) or 10 (DeltaN10-) residues removed from the N-terminal at 2.1-, 1.7-, and 1.9-A resolution, respectively. The crystal structures have C-alpha root-mean-squared differences of less than 0.8 A for the central core consisting of residues 11-98, showing that alterations of the N terminal have little influence on the folded core of the protein. The central core has the same fold topology as observed in the NMR structures of T4moD, the methane monooxygenase effector protein (MmoB) from two methanotrophs, and the phenol hydroxylase effector protein (DmpM). However, the root-mean-squared differences between comparable C-alpha positions in the X-ray structures and the NMR structures vary from approximately 1.8 A to greater than 6 A. The X-ray structures exhibit an estimated overall coordinate error from 0.095 (0.094) A based on the R-value (R free) for the highest resolution DeltaN4-T4moD structure to 0.211 (0.196) A for the native T4moD structure. Catalytic studies of the DeltaN4-, DeltaN7-, and DeltaN10- variants of T4moD show statistically insignificant changes in k(cat), K(M), k(cat)/K(M), and K(I) relative to the native protein. Moreover, there was no significant change in the regiospecificity of toluene oxidation with any of the T4moD variants. The relative insensitivity to changes in the N-terminal region distinguishes T4moD from the MmoB homologues, which each require the approximately 33 residue N-terminal region for catalytic activity. PMID- 15882053 TI - New binding specificities derived from Min-23, a small cystine-stabilized peptidic scaffold. AB - The randomization of both internal and surface residues in small protein domains followed by selection from a display library is emerging as a powerful strategy to obtain novel binding specificities. Small and stable scaffold motifs observed in disulfide-rich proteins are attractive because they are small, stable, and accessible to chemical synthesis. The elementary structural motif found in the squash trypsin inhibitor EETI-II (Ecballium elaterium trypsin inhibitor) is the cystine stabilized beta-sheet (CSB) motif, found in nearly 50% of all known small disulfide-rich protein families. We have used Min-23, a short 23-residue peptide containing the CSB motif and shown to be a stable autonomous folding unit and one of the smallest scaffolds described to date, as a scaffold for selection of new binding ligands. We demonstrate that the core CSB motif in Min-23 is permissive to loop insertion, using peptide epitopes from hemagglutinin (HA) and Gla-protein (E). A phage library of more than 10(8) different clones has been constructed by insertion of a randomized sequence on a beta-turn of the Min-23 peptide. The selection of this library on a variety of 7 different targets allowed the isolation of 21 new specific binders, confirming the potential of Min-23 as a scaffold for the development of new ligands. The derived library is able to provide a wide range of novel compounds with possible applications in various biological and pharmaceutical areas. PMID- 15882054 TI - Mutual information in protein multiple sequence alignments reveals two classes of coevolving positions. AB - Information theory was used to identify nonconserved coevolving positions in multiple sequence alignments from a variety of protein families. Coevolving positions in these alignments fall into two general categories. One set is composed of positions that coevolve with only one or two other positions. These positions often display direct amino acid side-chain interactions with their coevolving partner. The other set comprises positions that coevolve with many others and are frequently located in regions critical for protein function, such as active sites and surfaces involved in intermolecular interactions and recognition. We find that coevolving positions are more likely to change protein function when mutated than are positions showing little coevolution. These results imply that information theory may be applied generally to find coevolving, nonconserved positions that are part of functional sites in uncharacterized protein families. We propose that these coevolving positions compose an important subset of the positions in an alignment, and may be as important to the structure and function of the protein family as are highly conserved positions. PMID- 15882055 TI - Identification of an SH3-binding motif in a new class of methionine aminopeptidases from Mycobacterium tuberculosis suggests a mode of interaction with the ribosome. AB - The crystal structure of the methionine aminopeptidase (MetAP) from Mycobacterium tuberculosis (MtMetAP1c) has been determined in the apo- and methionine-bound forms. This is the first structure of a type I MetAP with a significant extension at the amino terminus. The catalytic domain is similar to that of Escherichia coli MetAP (EcMetAP), and the additional 40-residue segment wraps around the surface with an extended but well-defined structure. There are several members of the actinomyces family of bacteria that contain MetAPs with such N-terminal extensions, and we classify these as MetAP type Ic (MetAP1c). Some members of this family of bacteria also contain a second MetAP (type Ia) similar in size to EcMetAP. The main difference between the apo- and the methionine-bound forms of MtMetAP1c is in the conformation of the metal-binding residues. The position of the methionine bound in the active site is very similar to that found in many of the known members of this family. Side chains of several residues in the S1 and S1' subsites shift as much as 1.5 A compared to EcMetAP. Residues 14-17 have the sequence Pro-Thr-Arg-Pro and adopt the conformation of a polyproline II helix. Model-building suggests that this PxxP segment can bind to an SH3 protein motif. Other type Ib and type Ic MetAPs with N-terminal extensions contain similarly located PxxP motifs. Also, several ribosomal proteins are known to include SH3 domains, one of which is located close to the tunnel from which the nascent polypeptide chain exits the ribosome. Therefore, it is proposed that the binding of MetAPs to the ribosome is mediated by a complex between a PxxP motif on the protein and an SH3 domain on the ribosome. It is also possible that zinc-finger domains, which are located at the extreme N-terminus of type I MetAPs, may participate in interactions with the ribosome. PMID- 15882056 TI - Aromatic ring cleavage by homoprotocatechuate 2,3-dioxygenase: role of His200 in the kinetics of interconversion of reaction cycle intermediates. AB - Homoprotocatechuate 2,3-dioxygenase (WT 2,3-HPCD) isolated from Brevibacterium fuscum utilizes an active site Fe(II) and O(2) to catalyze proximal extradiol cleavage of the aromatic ring of the substrate (HPCA). Here, the conserved active site residue His200 is changed to Gln, Glu, Ala, Asn, and Phe, and the reactions of the mutant enzymes are probed using steady-state and transient kinetic techniques. Each mutant catalyzes ring cleavage of HPCA to yield the normal product. H200Q and H200N retain 30-40% of the WT 2,3-HPCD activity at 24 degrees C, but the other mutants reduce the k(cat) to less than 9% of normal. The origin of the reduced activity is unlikely to be the substrate binding phase of the catalytic cycle, because the multistep anaerobic binding reaction of the chromophoric substrate 4-nitrocatechol (4NC) is shown to proceed with rate constants similar to those observed for WT 2,3-HPCD. In contrast, the rate constants of several steps in the multistep O(2) binding/insertion and product release half of the reaction cycle are substantially slowed, in particular the steps in which activated oxygen attacks the organic substrate and in which product is released. In the case of the H200N mutant, the product of 4NC oxidation is not the usual ring cleavage product, but rather the 4NC quinone. These results suggest that the main role of His200 is in facilitating the steps in the second half of the reaction cycle. The decreased rate constants for the O(2) insertion steps in the catalytic cycles of the mutant enzymes allow the oxygen adduct of an extradiol dioxygenase to be detected for the first time. PMID- 15882057 TI - Accumulation of multiple intermediates in the catalytic cycle of (4 hydroxyphenyl)pyruvate dioxygenase from Streptomyces avermitilis. AB - (4-Hydroxyphenyl)pyruvate dioxygenase (HPPD) catalyzes the conversion of (4 hydroxyphenyl)pyruvate (HPP) to homogentisate (HG). This reaction involves decarboxylation, substituent migration, and aromatic oxygenation in a single catalytic cycle. HPPD is a unique member of the alpha-keto acid dependent oxygenases that require Fe(II) and an alpha-keto acid substrate to oxygenate or oxidize an organic molecule. We have examined the reaction coordinate of HPPD from Streptomyces avermitilis using rapid mixing pre-steady-state methods in conjunction with steady-state kinetic analyses. Acid quench reactions and product analysis of homogentisate indicate that HPPD as isolated is fully active and that experiments limited in dioxygen concentration with respect to that of the enzyme do involve a single turnover. These experiments indicate that during the course of one turnover the concentration of homogentisate is stoichiometric with enzyme concentration by approximately 200 ms, well before the completion of the catalytic cycle. Subsequent single turnover reactions were monitored spectrophotometrically under pseudo-first-order and matched concentration reactant conditions. Three spectrophotometrically distinct intermediates are observed to accumulate. The first of these is a relatively strongly absorbing species with maxima at 380 and 480 nm that forms with a rate constant (k(1)) of 7.4 x 10(4) M(-)(1) s(-)(1) and then decays to a second intermediate with a rate constant (k(2)) of 74 s(-)(1). The rate constant for the decay of the second intermediate (k(3)) is 13 s(-)(1) and is concomitant with the formation of the product, homogentisate, based on rapid quench and pre-steady-state fluorescence measurements. The rate constant for this process decreases to 7.6 s(-)(1) when deuterons are substituted for protons in the aromatic ring of the substrate. The release of product from the enzyme is rate limiting and occurs at 1.6 s(-)(1). This final event exhibits a kinetic isotope effect of 2 with deuterium oxide as the solvent, consistent with a solvent isotope effect on V(max) of 2.6 observed in steady-state experiments. PMID- 15882058 TI - Site-directed mutagenesis of proline 94 to alanine in amicyanin converts a true electron transfer reaction into one that is kinetically coupled. AB - Amicyanin is a type I copper protein that mediates electron transfer (ET) from methylamine dehydrogenase (MADH) to cytochrome c-551i. Pro(94) resides in the "ligand loop" of amicyanin, a sequence of amino acids that contains three of the four copper ligands. ET from the reduced O-quinol tryptophan tryptophylquinone of MADH to oxidized P94A amicyanin is a true ET reaction that exhibits values of electronic coupling (H(AB)) and reorganization energy (lambda) that are the same as for the reaction of native amicyanin. In contrast, the parameters for the ET reaction from reduced P94A amicyanin to oxidized cytochrome c-551i have been significantly altered as a consequence of the mutation. These values of H(AB) and lambda are 8.3 cm(-)(1) and 2.3 eV, respectively, compared to values of 0.3 cm( )(1) and 1.2 eV for the reaction of native reduced amicyanin. The crystal structure of reduced P94A amicyanin exhibits two alternate conformations with the positions of the copper 1.4 A apart [Carrell, C. J., Sun, D., Jiang, S., Davidson, V. L., and Mathews, F. S. (2004) Biochemistry 43, 9372-9380]. In one of these, conformation B, a water molecule has replaced Met(98) as a copper ligand, and the ET distance to the heme of the cytochrome is increased by 1.4 A. Analysis of these structures suggests that the true k(ET) for ET from the copper in conformation B to heme would be much less than for ET from conformation A. A novel kinetic mechanism is proposed to explain these data in which the reduction of Cu(2+) by methylamine dehydrogenase is a true ET reaction while the oxidation of Cu(1+) by cytochrome c-551i is kinetically coupled ET. By comparison of the temperature dependence of the observed rate of the coupled ET reaction from reduced P94A amicyanin to cytochrome c-551i with the predicted rates and temperature dependence for the true ET reaction from conformation A, it was possible to determine the K(eq) and values of DeltaH degrees and DeltaS degrees that are associated with the non-ET reaction that modulates the observed ET rate. PMID- 15882059 TI - A biophysical investigation of recombinant hemoglobins with aromatic B10 mutations in the distal heme pockets. AB - This study examines the structural and functional effects of amino acid substitutions in the distal side of both the alpha- and beta-chain heme pockets of human normal adult hemoglobin (Hb A). Using our Escherichia coli expression system, we have constructed four recombinant hemoglobins: rHb(alphaL29F), rHb(alphaL29W), rHb(betaL28F), and rHb(betaL28W). The alpha29 and beta28 residues are located in the B10 helix of the alpha- and beta-chains of Hb A, respectively. The B10 helix is significant because of its proximity to the ligand-binding site. Previous work showed the ability of the L29F mutation to inhibit oxidation. rHb(alphaL29W), rHb(betaL28F), and rHb(betaL28W) exhibit very low oxygen affinity and reduced cooperativity compared to those of Hb A, while the previously studied rHb(alphaL29F) exhibits high oxygen affinity. Proton nuclear magnetic resonance spectroscopy indicates that these mutations in the B10 helix do not significantly perturb the alpha(1)beta(1) and alpha(1)beta(2) subunit interfaces, while as expected, the tertiary structures near the heme pockets are affected. Experiments in which visible spectrophotometry was utilized reveal that rHb(alphaL29F) has equivalent or slower rates of autoxidation and azide-induced oxidation than does Hb A, while rHb(alphaL29W), rHb(betaL28F), and rHb(betaL28W) have increased rates. Bimolecular rate constants for NO-induced oxidation have been determined using a stopped-flow apparatus. These findings indicate that amino acid residues in the B10 helix of the alpha- and beta-chains can play different roles in regulating the functional properties and stability of the hemoglobin molecule. These results may provide new insights for designing a new generation of hemoglobin-based oxygen carriers. PMID- 15882060 TI - Mapping of steroids binding to 17 beta-hydroxysteroid dehydrogenase type 1 using Monte Carlo energy minimization reveals alternative binding modes. AB - Crystallographic studies of ligand-protein complexes reveal most preferable ligand binding modes, but do not show less populated modes that may contribute to measurable biochemical and biophysical characteristics of the complexes. In some cases, a ligand may bind a protein in essentially different modes. An example is 17beta-hydroxysteroid dehydrogenase type 1 (17beta-HSD1), a steroidogenic enzyme that catalyzes reduction of estrone to estradiol in gonadal and peripheral tissues. The enzyme exhibits a high specificity for estrogens which bind with their C17 atom in the proximity of the NADP(H) cofactor. 17Beta-HSD1 can also bind androgens, but in a reverse binding mode, in which the steroid C3 atom is the closest carbon atom to the cofactor. Here we map the interaction energy of estradiol and dihydrotestosterone binding to 17beta-HSD1. Positions and orientations of the steroids in the ligand-binding tunnel were sampled systematically, and at each combination of these generalized coordinates, the energy was Monte Carlo minimized. The computed maps show energy minima corresponding to the X-ray structures and predict alternative binding modes, in particular, an upside-down orientation in which steroidal face alpha is exposed to protein residues that normally interact with face beta. The methodology can be used for mapping ligand-receptor interactions in various systems, for example, in ion channels and G-protein-coupled receptors that bind elongated ligands in confined space between transmembrane helices. PMID- 15882061 TI - Domain motions and the open-to-closed conformational transition of an enzyme: a normal mode analysis of S-adenosyl-L-homocysteine hydrolase. AB - The structure and fluctuations of the enzyme S-adenosyl-L-homocysteine hydrolase (SAHH) are analyzed in an effort to explain its biological function. Besides the previously identified open structure, characteristic of the substrate-free enzyme, we find two distinct structures in enzyme-inhibitor complexes, the closed and closed-twisted conformers. Both closed conformers differ from the open form by a hinge bending motion of two large domains within each subunit, which isolate the inhibitor bound in the active site from the bulk solvent. The closed-twisted form further differs from the closed form by a rigid body twist of the two subunit dimers. The local structural fluctuations of SAHH are analyzed by performing block normal mode analysis of the tetrameric enzyme in its three forms. For the open form, we find that the four lowest-frequency normal modes, corresponding to the collective motions of the protein with the largest amplitudes, are essentially combinations of the hinge bending deformations of the individual subunits. Thus, the mechanical properties of the open structure of SAHH lead to the presence of structural fluctuations in the direction of the open to-closed conformational transition. A candidate for such a motion has been observed in previous fluorescence depolarization studies of the enzyme. Both structural and normal mode analyses indicate that residues 180-190 and 350-356 form hinge regions, connecting large domains which tend to move as rigid bodies in response to interactions with substrate, intermediates, and the product of the enzymatic reactions. We propose that these hinge regions play a crucial role in the enzymatic mechanism of SAHH. In contrast to the open form, normal mode calculations for the closed conformations show strong coupling of the hinge bending motions of the individual subunits to each other and to other low frequency vibrations. Thus, information about structural changes related to reaction progress in one active site may be mechanically transmitted to other subunits of the protein, explaining the cooperativity found in the enzyme kinetics. PMID- 15882062 TI - Domain-domain communication for tRNA aminoacylation: the importance of covalent connectivity. AB - Aminoacyl-tRNA synthetases form complexes with tRNA to catalyze transfer of activated amino acids to the 3' end of tRNA. The tRNA synthetase complexes are roughly divided into the activation and tRNA-binding domains of synthetases, which interact with the acceptor and anticodon ends of tRNAs, respectively. Efficient aminoacylation of tRNA by Escherichia coli cysteinyl-tRNA synthetase (CysRS) requires both domains, although the pathways for the long-range domain domain communication are not well understood. Previous studies show that dissection of tRNA(Cys) into acceptor and anticodon helices seriously reduces the efficiency of aminoacylation, suggesting that communication requires covalent continuity of the tRNA backbone. Here we tested if communication requires the continuity of the synthetase backbone. Two N-terminal fragments and one C terminal fragment of E. coli CysRS were generated. While the N-terminal fragments were active in adenylate synthesis, they were severely defective in the catalytic efficiency and specificity of tRNA aminoacylation. Conversely, although the C terminal fragment was not catalytically active, it was able to bind and discriminate tRNA. However, addition of the C-terminal fragment to an N-terminal fragment in trans did not improve the aminoacylation efficiency of the N-terminal fragment to the level of the full-length enzyme. These results emphasize the importance of covalent continuity of both CysRS and tRNA(Cys) for efficient tRNA aminoacylation, and highlight the energetic costs of constraining the tRNA synthetase complex for domain-domain communication. Importantly, this study also provides new insights into the existence of several natural "split" synthetases that are now identified from genomic sequencing projects. PMID- 15882063 TI - Induced fit in HIV-neutralizing antibody complexes: evidence for alternative conformations of the gp120 V3 loop and the molecular basis for broad neutralization. AB - Human monoclonal antibody (mAb) 447-52D neutralizes a broad spectrum of HIV-1 isolates, whereas murine mAb 0.5beta, raised against gp120 of the X4 isolate HIV 1(IIIB), neutralizes this strain specifically. Two distinct gp120 V3 peptides, V3(MN) and V3(IIIB), adopt alternative beta-hairpin conformations when bound to 447-52D and 0.5beta, respectively, suggesting that the alternative conformations of this loop play a key role in determining the coreceptor specificity of HIV-1. To test this hypothesis and to better understand the molecular basis underlying an antibody's breadth of neutralization, the solution structure of the V3(IIIB) peptide bound to 447-52D was determined by NMR. V3(IIIB) and V3(MN) peptides bound to 447-52D exhibited the same N-terminal strand conformation, while the V3(IIIB) peptide revealed alternative N-terminal conformations when bound to 447 52D and 0.5beta. Comparison of the three known V3 structures leads to a model in which a 180 degrees change in the orientation of the side chains and the resulting one-residue shift in hydrogen bonding patterns in the N-terminal strand of the beta-hairpins markedly alter the topology of the surface that interacts with antibodies and that can potentially interact with the HIV-1 coreceptors. Predominant interactions of 447-52D with three conserved residues of the N terminal side of the V3 loop, K312, I314, and I316, can account for its broad cross reactivity, whereas the predominant interactions of 0.5beta with variable residues underlie its strain specificity. PMID- 15882064 TI - Novel conotoxins from Conus striatus and Conus kinoshitai selectively block TTX resistant sodium channels. AB - The peptides isolated from venoms of predatory marine Conus snails ("conotoxins") are well-known to be highly potent and selective pharmacological agents for voltage-gated ion channels and receptors. We report the discovery of two novel TTX-resistant sodium channel blockers, mu-conotoxins SIIIA and KIIIA, from two species of cone snails. The two toxins were identified and characterized by combining molecular techniques and chemical synthesis. Both peptides inhibit TTX resistant sodium currents in neurons of frog sympathetic and dorsal root ganglia but poorly block action potentials in frog skeletal muscle, which are mediated by TTX-sensitive sodium channels. The amino acid sequences in the C-terminal region of the two peptides and of the previously characterized mu-conotoxin SmIIIA (which also blocks TTX-resistant channels) are similar, but the three peptides differ in the length of their first N-terminal loop. We used molecular dynamics simulations to analyze how altering the number of residues in the first loop affects the overall structure of mu-conotoxins. Our results suggest that the naturally occurring truncations do not affect the conformation of the C-terminal loops. Taken together, structural and functional differences among mu-conotoxins SmIIIA, SIIIA, and KIIIA offer a unique insight into the "evolutionary engineering" of conotoxin activity. PMID- 15882065 TI - Aggregation of a slow-folding mutant of a beta-clam protein proceeds through a monomeric nucleus. AB - Mechanistic understanding of protein aggregation, leading either to structured amyloid fibrils or to amorphous inclusion body-like deposits, should facilitate the identification of potential therapeutic intervention strategies for the devastating amyloid-based diseases. Here we focus on the in vitro aggregation of a slow-folding mutant of the beta-clam protein, cellular retinoic acid-binding protein I (P39A CRABP I), which forms inclusion bodies when expressed in Escherichia coli. Aggregation was monitored by observing the fluorescence of a fluorescein-based biarsenical dye (FlAsH) that ligates to a tetra-Cys motif, here incorporated into a flexible Omega-loop. The fluorescence signal of FlAsH on the tetra-Cys-containing P39A CRABP I is sensitive to whether this protein is native or unfolded, and was used in combination with other techniques to follow aggregate formation. The aggregation time course is compatible with a nucleation dependent polymerization model, and detailed kinetic analysis showed that the energetically unfavorable nucleus is monomeric. A similar conclusion was reached previously for poly(Gln) species [Chen, S., Ferrone, F. A., and Wetzel, R. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 11884-11889] and points to an unfavorable equilibrium between the misfolded intermediate and the bulk pool of monomers as causative in aggregation. The P39A mutation, which removes a helix-stop signal, may slow closure of the beta-barrel in P39A CRABP I relative to the wild type, leaving it vulnerable to aggregation. Wide-angle X-ray scattering showed that the amorphous aggregates formed by the aggregation-prone intermediates of P39A CRABP I contain predominantly beta-strands structured in a lamellar fashion with 10.03 A spacing between adjacent beta-sheets. PMID- 15882066 TI - Interactions between beta D372 and gamma subunit N-terminus residues gamma K9 and gamma S12 are important to catalytic activity catalyzed by Escherichia coli F1F0 ATP synthase. AB - Substitution of Escherichia coli F(1)F(0) ATP synthase residues betaD372 or gammaS12 with groups that are unable to form a hydrogen bond at this location decreased ATP synthase-dependent cell growth by 2 orders of magnitude, eliminated the ability of F(1)F(0) to catalyze ATPase-dependent proton pumping in inverted E. coli membranes, caused a 15-20% decrease in the coupling efficiency of the membranes as measured by the extent of succinate-dependent acridine orange fluorescence quenching, but increased soluble F(1)-ATPase activity by about 10%. Substitution of gammaK9 to eliminate the ability to form a salt bridge with betaD372 decreased soluble F(1)-ATPase activity and ATPase-driven proton pumping by 2-fold but had no effect on the proton gradient induced by addition of succinate. Mutations to eliminate the potential to form intersubunit hydrogen bonds and salt bridges between other less highly conserved residues on the gamma subunit N-terminus and the beta subunits had little effect on ATPase or ATP synthase activities. These results suggest that the betaD372-gammaK9 salt bridge contributes significantly to the rate-limiting step in ATP hydrolysis of soluble F(1) while the betaD372-gammaS12 hydrogen bond may serve as a component of an escapement mechanism for ATP synthesis in which alphabetagamma intersubunit interactions provide a means to make substrate binding a prerequisite of proton gradient-driven gamma subunit rotation. PMID- 15882067 TI - Structural characterization of the antimicrobial peptide pleurocidin from winter flounder. AB - Pleurocidin is an antimicrobial peptide that was isolated from the mucus membranes of winter flounder (Pseudopleuronectes americanus) and contributes to the initial stages of defense against bacterial infection. From NMR structural studies with the uniformly (15)N-labeled peptide, a structure of pleurocidin was determined to be in a random coil conformation in aqueous solution whereas it assumes an alpha-helical structure in TFE and in dodecylphosphocholine (DPC) micelles. From (15)N relaxation studies, the helix is a rigid structure in the membrane-mimicking environment. Strong NOESY cross-peaks from the pleurocidin to the aliphatic chain on DPC confirm that pleurocidin is contained within the DPC micelle and not associated with the surface of the micelle. From diffusion studies it was determined that each micelle contains at least two pleurocidin molecules. PMID- 15882068 TI - Combined interaction of phospholipase C and apolipoprotein A-I with small unilamellar lecithin-cholesterol vesicles: influence of apolipoprotein A-I concentration and vesicle composition. AB - We report the combined effects of phospholipase C (PLC), a pronucleating factor, and apolipoprotein A-I (apo A-I), an antinucleating factor, in solutions of model bile. Results indicate that apo A-I inhibits cholesterol nucleation from unilamellar lecithin vesicles by two mechanisms. Initially, inhibition is achieved by apo A-I shielding of hydrophobic diacylglycerol (DAG) moieties so as to prevent vesicle aggregation. Protection via shielding is temporary. It is lost when the DAG/apo A-I molar ratio exceeds a critical value. Subsequently, apo A-I forms small ( approximately 5-15 nm) complexes with lecithin and cholesterol that coexist with lipid-stabilized (400-800 nm) DAG oil droplets. This microstructural transition from vesicles to complexes avoids nucleation of cholesterol crystals and is a newly discovered mechanism by which apo A-I serves as an antinucleating agent in bile. The critical value at which a microstructural transition occurs depends on binding of apo A-I and so varies with the cholesterol mole fraction of vesicles. Aggregation of small, unilamellar, egg lecithin vesicles (SUVs) with varying cholesterol composition (0-60 mol %) was monitored for a range of apo A-I concentrations (2 to 89 microg/mL). Suppression of aggregation persists so long as the DAG-to-bound-apo A-I molar ratio is less than 100. A fluorescence assay involving dansylated lecithin shows that the suppression is an indirect effect of apo A-I rather than a direct inhibition of PLC enzyme activity. The DAG-to-total apo A-I molar ratio at which suppression is lost increases with cholesterol because of differences in apo A-I binding. Above this value, a microstructural transition to DAG droplets and lecithin/cholesterol A-I complexes occurs, as evidenced by sudden increases in turbidity and size and enhancement of Forster resonance energy transfer; structures are confirmed by cryo TEM. PMID- 15882069 TI - Calcium-binding properties of wild-type and EF-hand mutants of S100B in the presence and absence of a peptide derived from the C-terminal negative regulatory domain of p53. AB - S100B is a dimeric Ca(2+)-binding protein that undergoes a 90 +/- 3 degrees rotation of helix 3 in the typical EF-hand domain (EF2) upon the addition of calcium. The large reorientation of this helix is a prerequisite for the interaction between each subunit of S100B and target proteins such as the tumor suppressor protein, p53. In this study, Tb(3+) was used as a probe to examine how binding of a 22-residue peptide derived from the C-terminal regulatory domain of p53 affects the rate of Ca(2+) ion dissociation. In competition studies with Tb(3+), the dissociation rates of Ca(2+) (k(off)) from the EF2 domains of S100B in the absence and presence of the p53 peptide was determined to be 60 and 7 s( )(1), respectively. These data are consistent with a previously reported result, which showed that that target peptide binding to S100B enhances its calcium binding affinity [Rustandi et al. (1998) Biochemistry 37, 1951-1960]. The corresponding Ca(2+) association rate constants for S100B, k(on), for the EF2 domains in the absence and presence of the p53 peptide are 1.1 x 10(6) and 3.5 x 10(5) M(-)(1) s(-)(1), respectively. These two association rate constants are significantly below the diffusion control ( approximately 10(9) M(-)(1) s(-)(1)) and likely involve both Ca(2+) ion association and a Ca(2+)-dependent structural rearrangement, which is slightly different when the target peptide is present. EF hand calcium-binding mutants of S100B were engineered at the -Z position (EF-hand 1, E31A; EF-hand 2, E72A; both EF-hands, E31A + E72A) and examined to further understand how specific residues contribute to calcium binding in S100B in the absence and presence of the p53 peptide. PMID- 15882070 TI - Mechanism of zinc coordination by point-mutated structures of the distal CCHC binding motif of the HIV-1 NCp7 protein. AB - The kinetics of Zn(2+) binding by two point-mutated forms of the HIV-1 NCp7 C terminal zinc finger, each containing tridentate binding motif HCC [Ser49(35 50)NCp7] or CCC [Ala44(35-50)NCp7], has been studied by stopped-flow spectrofluorimetry. Both the formation and dissociation rate constants of the complexes between Zn(2+) and the two model peptides depend on pH. The results are interpreted on the basis of a multistep reaction model involving three Zn(2+) binding paths due to three deprotonated states of the coordinating motif, acting as monodentate, bidentate, and tridentate ligands. For Ser49(35-50)NCp7 around neutral pH, binding preferentially occurs via the deprotonated Cys36 in the bidentate state also involving His44. The binding rate constants for the monodentate and bidentate states are 1 x 10(6) and 3.9 x 10(7) M(-)(1) s(-)(1), respectively. For Ala44(35-50)NCp7, intermolecular Zn(2+) binding predominantly occurs via the deprotonated Cys36 in the monodentate state with a rate constant of 3.6 x 10(7) M(-)(1) s(-)(1). In both mutants, the final state of the Zn(2+) complex is reached by subsequent stepwise ligand deprotonation and intramolecular substitution of coordinated water molecules. The rate constants for the intermolecular binding paths of the bidentate and tridentate states of Ala44(35 50)NCp7 and of the tridentate state of Ser49(35-50)NCp7 are much smaller than expected according to electrostatic considerations. This is attributed to conformational constraints required to achieve proper metal coordination during folding. The dissociation of Zn(2+) from both peptides is again characterized by a multistep process and takes place fastest via the protonated Zn(2+)-bound bidentate and monodentate states, with rate constants of approximately 0.3 and approximately 10(3) s(-)(1), respectively, for Ser49(35-50)NCp7 and approximately 4 x 10(-)(3) and approximately 500 s(-)(1), respectively, for Ala44(35-50)NCp7. PMID- 15882071 TI - The use of (E)- and (Z)-phosphoenol-3-fluoropyruvate as mechanistic probes reveals significant differences between the active sites of KDO8P and DAHP synthases. AB - The enzymes 3-deoxy-d-manno-2-octulosonate-8-phosphate (KDO8P) synthase and 3 deoxy-d-arabino-2-heptulosonate-7-phosphate (DAHP) synthase catalyze a similar aldol-type condensation between phosphoenolpyruvate (PEP) and the corresponding aldose: arabinose 5-phosphate (A5P) and erythrose 4-phosphate (E4P), respectively. While KDO8P synthase is metal-dependent in one class of organisms and metal-independent in another, only a metal-dependent class of DAHP synthases has thus far been identified in nature. We have used catalytically active E and Z isomers of phosphoenol-3-fluoropyruvate [(E)- and (Z)-FPEP, respectively] as mechanistic probes to characterize the differences and/or the similarities between the metal-dependent and metal-independent KDO8P synthases as well as between the metal-dependent KDO8P synthase and DAHP synthase. The direct evidence of the overall stereochemistry of the metal-dependent Aquifex pyrophilus KDO8P synthase (ApKDO8PS) reaction was obtained by using (E)- and (Z)-FPEPs as alternative substrates and by subsequent (19)F NMR analysis of the products. The results reveal the si face addition of the PEP to the re face of the carbonyl of A5P, and establish that the stereochemistry of ApKDO8PS is identical to that of the metal-independent Escherichia coli KDO8P synthase enzyme (EcKDO8PS). In addition, both ApKDO8PS and EcKDO8PS enzymes exhibit high selectivity for (E) FPEP versus (Z)-FPEP, the relative k(cat)/K(m) ratios being 100 and 33, respectively. In contrast, DAHP synthase does not discriminate between (E)- and (Z)-FPEP (the k(cat)/K(m) being approximately 7 x 10(-)(3) microM(-)(1) s(-)(1) for both compounds). The pre-steady-state burst experiments for EcKDO8PS showed that product release is rate-limiting for the reactions performed with either PEP, (E)-FPEP, or (Z)-FPEP, although the rate constants, for both product formation and product release, were lower for the fluorinated analogues than for PEP [125 and 2.3 s(-)(1) for PEP, 2.5 and 0.2 s(-)(1) for (E)-FPEP, and 9 and 0.1 s(-)(1) for (Z)-FPEP, respectively]. The observed data indicate substantial differences in the PEP subsites and open the opportunity for the design of selective inhibitors against these two families of enzymes. PMID- 15882072 TI - The calcium-sensor guanylate cyclase activating protein type 2 specific site in rod outer segment membrane guanylate cyclase type 1. AB - The rod outer segment membrane guanylate cyclase type 1 (ROS-GC1), originally identified in the photoreceptor outer segments, is a member of the subfamily of Ca(2+)-modulated membrane guanylate cyclases. In phototransduction, its activity is tightly regulated by its two Ca(2+)-sensor protein parts, GCAP1 and GCAP2. This study maps the GCAP2-modulatory site in ROS-GC1 through the use of multiple techniques involving surface plasmon resonance binding studies with soluble ROS GC1 constructs, coimmunoprecipitation, functional reconstitution experiments with deletion mutants, and peptide competition assays. The findings show that the sequence motif of the core GCAP2-modulatory site is Y965-N981 of ROS-GC1. The site is distinct from the GCAP1-modulatory site. It, however, partially overlaps with the S100B-regulatory site. This indicates that the Y965-N981 motif tightly controls the Ca(2+)-dependent specificity of ROS-GC1. Identification of the site demonstrates an intriguing topographical feature of ROS-GC1. This is that the GCAP2 module transmits the Ca(2+) signals to the catalytic domain from its C terminal side and the GCAP1 module from the distant N-terminal side. PMID- 15882073 TI - Phosphorylation of apolipoprotein-E at an atypical protein kinase CK2 PSD/E site in vitro. AB - Apolipoprotein-E (apoE) plays an important role in neuronal lipid transport and is thought to stabilize microtubules by preventing tau hyperphosphorylation. ApoE is also associated with insoluble amyloid detected in Alzheimer disease brain lesions. The apoE C-terminal shares several physicochemical features with alpha synuclein, another neuronal apolipoprotein-like protein. Alpha-synuclein is phosphorylated by protein kinase CK2 (CK2) at an atypical PSD/E motif in vivo and in vitro. We identified a similar PSD/E motif in apoE and therefore investigated its potential phosphorylation by CK2 in vitro. When a [(32)P]-labeling approach was used, CK2 readily phosphorylated purified human apoE as well as recombinant forms of human apoE3 and apoE4. Using liquid chromatography mass spectrometry techniques, we mapped the major apoE CK2 phosphorylation site to Ser296 within the apoE PSD/E motif. We also found that apoE potently activated CK2 as demonstrated by increased CK2beta subunit autophosphorylation and by increased phosphorylation of tau when the latter was added to the kinase reaction mixtures. Other proteins such as apolipoprotein A-I and albumin did not effectively activate CK2. The phosphorylation of apoE by CK2 as well as the activation of CK2 by apoE may be relevant in vivo where apoE, CK2, and tau are co-localized with additional CK2 targets on neuronal microtubules. PMID- 15882074 TI - Characteristics of iodide activation and inhibition of oxygen evolution by photosystem II. AB - Oxygen evolution by photosystem II (PSII) is activated by chloride and other monovalent anions. In this study, the effects of iodide on oxygen evolution activity were investigated using PSII-enriched membrane fragments from spinach. In the absence of Cl(-), the dependence of oxygen evolution activity on I(-) concentration showed activation followed by inhibition in both intact PSII and NaCl-washed PSII, which lacked the PsbP and PsbQ subunits. Using a substrate inhibition model, the range of values of the Michaelis constant K(M) in intact PSII (0.5-1.5 mM) was smaller than that in NaCl-washed PSII (1.5-5 mM), whereas values of the inhibition constant K(I) in intact PSII (9-17 mM) were larger than those in NaCl-washed PSII (1-4 mM). Studies of I(-) inhibition of Cl(-)-activated oxygen evolution in intact PSII revealed that I(-) was primarily an uncompetitive inhibitor, with uncompetitive constant K(i)' = 37 mM and Cl(-)-competitive constant K(i) > 200 mM. This result indicated that the activating Cl(-) must be bound for inhibition to take place, which is consistent with the substrate inhibition model for I(-) activation. The S(2) state multiline and g = 4.1 EPR signals in NaCl-washed PSII were examined in the presence of 3 and 25 mM NaI, corresponding to I(-)-activated and I(-)-inhibited conditions, respectively. The two S(2) state signals were observed at both I(-) concentrations, indicating that I(-) substitutes for Cl(-) in formation of the signals and that advancement to the S(2) state was not prevented by high I(-) concentrations. A model is presented that incorporates the results of this study, including the action of both chloride and iodide. PMID- 15882075 TI - Cooperative interaction of human XPA stabilizes and enhances specific binding of XPA to DNA damage. AB - Human xeroderma pigmentosum group A (XPA) is an essential protein for nucleotide excision repair (NER). We have previously reported that XPA forms a homodimer in the absence of DNA. However, what oligomeric forms of XPA are involved in DNA damage recognition and how the interaction occurs in terms of biochemical understanding remain unclear. Using the homogeneous XPA protein purified from baculovirus-infected Sf21 insect cells and the methods of gel mobility shift assays, gel filtration chromatography, and UV-cross-linking, we demonstrated that both monomeric and dimeric XPA bound to the DNA adduct of N-acetyl-2 aminofluorene (AAF), while showing little affinity for nondamaged DNA. The binding occurred in a sequential and protein concentration-dependent manner. At relatively low-protein concentrations, XPA formed a complex with DNA adduct as a monomer, while at the higher concentrations, an XPA dimer was involved in the specific binding. Results from fluorescence spectroscopic and competitive binding analyses indicated that the specific binding of XPA to the adduct was significantly facilitated and stabilized by the presence of the second XPA in a positive cooperative manner. This cooperative binding exhibited a Hill coefficient of 1.9 and the step binding constants of K(1) = 1.4 x 10(6) M(-)(1) and K(2) = 1.8 x 10(7) M(-)(1). When interaction of XPA and RPA with DNA was studied, even though binding of RPA-XPA complex to adducted DNA was observed, the presence of RPA had little effect on the overall binding efficiency. Our results suggest that the dominant form for XPA to efficiently bind to DNA damage is the XPA dimer. We hypothesized that the concentration-dependent formation of different types of XPA-damaged DNA complex may play a role in cellular regulation of XPA activity. PMID- 15882076 TI - Substrate-induced tryptophan fluorescence changes in EmrE, the smallest ion coupled multidrug transporter. AB - Tryptophan residues may play several roles in integral membrane proteins including direct interaction with substrates. In this work we studied the contribution of tryptophan residues to substrate binding in EmrE, a small multidrug transporter of Escherichia coli that extrudes various positively charged drugs across the plasma membrane in exchange with protons. Each of the four tryptophan residues was replaced by site-directed mutagenesis. The only single substitutions that affected the protein's activity were those in position 63. While cysteine and tyrosine replacements yielded a completely inactive protein, the replacement of Trp63 with phenylalanine brought about a protein that, although it could not confer any resistance against the toxicants tested, could bind substrate with an affinity 2 orders of magnitude lower than that of the wild-type protein. Double or multiple cysteine replacements at the other positions generate proteins that are inactive in vivo but regain their activity upon solubilization and reconstitution. The findings suggest a possible role of the tryptophan residues in folding and/or insertion. Substrate binding to the wild-type protein and to a mutant with a single tryptophan residue in position 63 induced a very substantial fluorescence quenching that is not observed in inactive mutants or chemically modified protein. The reaction is dependent on the concentration of the substrate and saturates at a concentration of 2.57 microM with the protein concentration of 5 microM supporting the contention that the functional unit is a dimer. These findings strongly suggest the existence of an interaction between Trp63 and substrate, and the nature of this interaction can now be studied in more detail with the tools developed in this work. PMID- 15882077 TI - Kinetic analysis of the role of histidine chloramines in hypochlorous acid mediated protein oxidation. AB - Hypochlorous acid (HOCl) is a powerful oxidant generated from H(2)O(2) and chloride ions by the heme enzyme myeloperoxidase (MPO) released from activated leukocytes. In addition to its potent antibacterial effects, excessive HOCl production can lead to host tissue damage, with this implicated in human diseases such as atherosclerosis, cystic fibrosis, and arthritis. HOCl reacts rapidly with biological materials, with proteins being major targets. Chlorinated amines (chloramines) formed from Lys and His side chains and alpha-amino groups on proteins are major products of these reactions; these materials are however also oxidants and can undergo further reactions. In this study, the kinetics of reaction of His side-chain chloramines with other protein components have been investigated by UV/visible spectroscopy and stopped flow methods at pH 7.4 and 22 degrees C, using the chloramines of the model compound 4-imidazoleacetic acid and N-alpha-acetyl-histidine. The second-order rate constants decrease in a similar order (Cys > Met > disulfide bonds > Trp approximately alpha-amino > Lys >> Tyr > backbone amides > Arg) to the corresponding reactions of HOCl, but are typically 5-25 times slower. These rate constants are consistent with His side-chain chloramines being important secondary oxidants in HOCl-mediated damage. These studies suggest that formation and subsequent reactions of His side-chain chloramines may be responsible for the targeted secondary modification of selected protein residues by HOCl that has previously been observed experimentally and highlight the importance of chloramine structure on their subsequent reactivity. PMID- 15882080 TI - Attainment of personal outcomes by people with developmental disabilities. AB - In 1993, The Council on Quality and Leadership in Supports for People With Disabilities (The Council) developed a person-centered outcome assessment instrument, Personal Outcome Measures (1997). The Council has maintained a database (N = 3,630) of Personal Outcome Measures interviews conducted during accreditation reviews. Our purpose in this article is to report findings on (a) the relationships between and among the Personal Outcome Measures (The Council, 1997) and patterns of individual outcome attainment and (b) to examine the impact of characteristic variables (disability, organizational size, residential type, or source of program funding) on the individual attainment of personal outcomes. Alternatives are suggested for examining the dimensions of both quality of service and quality of life for people with disabilities. PMID- 15882081 TI - Health status, utilization patterns, and outcomes of persons with intellectual disabilities: review of the literature. AB - A literature review was conducted to analyze existing data on health status, health care utilization, and medical outcomes of persons with intellectual disabilities. We found that barriers exist, but evidence indicates that (a) individuals with intellectual disability and medical needs are living in the community, (b) people with significant medical conditions can be maintained in community settings, and (c) medical supports can and are provided to people with intellectual disabilities and allied medical conditions to enable community living. However, some people had unmet medical needs, and the availability of and access to community-based services were problematic for others. These contradictory findings indicate that it is possible to provide services in the community, but the service delivery system needs improvement. PMID- 15882082 TI - Postnatal support for mothers of children with Down syndrome. AB - Delivering and receiving a postnatal diagnosis of Down syndrome is not an easy experience for most physicians or parents. In this study, 467 mothers of children with Down syndrome in Spain completed a survey about the postnatal support services they received immediately following the diagnosis of their child. Mothers reported feeling anxious, frightened, guilty, angry, and, in rare cases, suicidal. According to most mothers, physicians did not give adequate amounts of information about Down syndrome and rarely did they give enough printed materials or make referrals to parent support groups. Little seems to have changed since 1972. Mothers provided recommendations on how the Spanish medical system could be improved, with implications for other countries including the United States. PMID- 15882083 TI - Parents' perspectives on institutions. PMID- 15882084 TI - Outcomes that matter: parents' perspectives. PMID- 15882086 TI - Bonding corneal tissue: applications of photoactivated diazopyruvoyl cross linking agent. AB - Photoactivated bis-diazopyruvamide-N,N'-bis(3-diazopyruvoyl)-2,2' (ethylenedioxy)bis-(ethylamine), (DPD)-was previously shown to bond materials containing type I collagen. However, tensile strength of bonded collagenous tissue ( approximately 78% water) was low compared with that of dehydrated collagenous gelatin ( approximately 14% water). Here we investigated the role of water in corneal tissue bond strength and in bonding corneal tissue to glass. Bonding corneal tissue to glass may be of value in surgically anchoring keratoprostheses to corneas to alleviate problems with extrusion. Bovine corneal samples were lyophilized for various times resulting in tissue hydrations of zero (no water content) to approximately 3.7 (normal water content). The lyophilized corneal tissue was bonded to solid gelatin sheets, to other corneal samples and to glass using 0.3M DPD in chloroform. Control runs used chloroform only. Samples were irradiated with 100 or 200 J of 320-500 nm light. Strong bonds formed with all three materials when corneal tissue hydration was 1. No bonding occurred with chloroform alone. Formation of strong bonds only occurs with hydration levels Asn) mutant to possess greatly reduced efficiencies in catalysis of both chemical steps (alkylation and hydrolysis). PMID- 15882149 TI - An olive pollen protein with allergenic activity, Ole e 10, defines a novel family of carbohydrate-binding modules and is potentially implicated in pollen germination. AB - CBMs (carbohydrate-binding modules) are the most common non-catalytic modules associated with enzymes active in plant cell-wall hydrolysis. They have been frequently identified by amino acid sequence alignments, but only a few have been experimentally established to have a carbohydrate-binding activity. A small olive pollen protein, Ole e 10 (10 kDa), has been described as a major inducer of type I allergy in humans. In the present study, the ability of Ole e 10 to bind several polysaccharides has been analysed by affinity gel electrophoresis, which demonstrated that the protein bound 1,3-beta-glucans preferentially. Analytical ultracentrifugation studies confirmed binding to laminarin, at a protein/ligand ratio of 1:1. The interaction of Ole e 10 with laminarin induced a conformational change in the protein, as detected by CD and fluorescence analyses, and an increase of 3.6 degrees C in the thermal denaturation temperature of Ole e 10 in the presence of the glycan. These results, and the absence of alignment of the sequence of Ole e 10 with that of any classified CBM, indicate that this pollen protein defines a novel family of CBMs, which we propose to name CBM43. Immunolocalization of Ole e 10 in mature and germinating pollen by transmission electron microscopy and confocal laser scanning microscopy demonstrated the co localization of Ole e 10 and callose (1,3-beta-glucan) in the growing pollen tube, suggesting a role for this protein in the metabolism of carbohydrates and in pollen tube wall re-formation during germination. PMID- 15882150 TI - Usefulness of embolic protection devices during saphenous vein graft intervention in a nonselected population. AB - Distal embolic protection devices have been shown to reduce the risk of peri procedural complications associated with saphenous vein graft intervention. However, there are several important anatomic limitations that may preclude routine device use. We analyzed the angiograms of 624 consecutive saphenous vein graft interventions to determine eligibility for use of a proximal or distal embolic protection system. Overall, 483 (77%) of cases had anatomic and lesion characteristics suitable for an embolic protection device, including 59% eligible for proximal protection and 57% for distal protection. PMID- 15882151 TI - Protecting the patient from saphenous vein graft intervention. PMID- 15882152 TI - The embolic protection toolbox. PMID- 15882153 TI - Recovery of left ventricular systolic function after left anterior descending coronary artery stenting. AB - Our objective was to determine the factors affecting recovery of left ventricular (LV) contractility and myocardial perfusion after percutaneous coronary intervention (PCI). We selected 60 consecutive patients who underwent successful left anterior descending coronary artery (LAD) stenting. The mean stent diameter and length were 3.37+/-0.47 mm and 17.4+/-6 mm, respectively. Supporting a functional impact of successful PCI, myocardial perfusion and LV ejection fraction (LVEF) improved at 6+/-3 months after the procedure (48.8+/-11.6% vs 52.5+/-11.5%, P=0.05). Patient related factors such as diabetes mellitus, presentation with acute coronary syndrome, and age did not seem to affect LVEF change after the procedure. On univariate analysis, the change in LVEF after PCI was only related to the stent diameter. The increase in LVEF was higher in patients who received a stent>3 mm in diameter (P=0.041). There was a weak but statistically positive correlation between the stent diameter and the LVEF change after the procedure (R=0.267, P=0.049). Other procedure related factors such as multivessel PCI or stent length did not affect the percent ejection fraction change after stenting. PMID- 15882154 TI - Percutaneous closure of congenital and acquired ventricular septal defects- considerations on selection of the occlusion device. AB - Nonsurgical closure of congenital and acquired ventricular septal defects (VSD) has become increasingly acceptable with the availability of various occlusion systems that allow percutaneous treatment of muscular and membranous defects. This study describes a series of 12 patients (0.2-74-years-old) who underwent defect closure with six different occlusion systems. Device selection according to anatomy and outcome is highlighted. Seven VSDs were located in the membranous part of the septum, five in the mid-muscular septum. Complex heart lesions were present in five postmyocardial infarction VSD in one and residual postsurgical defects in three patients. The size of the VSD ranged from 2.6 to 10 mm. The applied devices were: Amplatzer muscular VSD occluder (n=4), Amplatzer septal occluder (n=2), Amplatzer duct occluder (n=1), Amplatzer membranous VSD occluder (n=2), Nit-Occlud coil (n=2), and Cook PDA coil (n=1). The devices were implanted successfully in nine patients. There was complete VSD closure in eight patients within the first 24 hours. In one patient, a trivial residual shunt disappeared at 6 months follow-up. Because of device instability, two occluders were removed during catheterization. In two other cases, tricuspid valve tissue was entrapped in the occluder and had to be removed surgically, one of them during the consecutive Rastelli operation. Neither significant arrhythmia, nor thromboembolism or hemolysis occurred in out patients during follow-up. Transcatheter closure of VSD is an attractive alternative to surgery. In complex congenital heart disease, surgical-interventional hybrid therapy may improve morbidity and total outcome. The recently developed Amplatzer VSD devices allow closure of muscular and membranous VSDs. Implantation and short-term follow-up are superior to the formerly used devices. Long-term effects have to be evaluated in further studies. PMID- 15882155 TI - Anomalous origin of right coronary artery from the left coronary sinus: incidence, characteristics, and a systematic approach for rapid diagnosis. AB - Twenty-three patients were found to have anomalous origin of right coronary artery from the left coronary sinus (LCS) from January 2000 to October 2003. The mean age was 58.6+/-14.3 years with male predominance (56.5%). Cardiovascular risk factors were found in 18 (78.3%) patients while coronary artery disease was seen in 13 (56.5%) patients. Among the coronary artery disease patients, the left anterior descending artery was the most commonly involved, followed by the right coronary artery and the left circumflex artery. Right coronary artery dominance was seen in 19 (82.6%) patients. The anomalous right coronary artery originates within the left coronary sinus in 17 (73.9%) patients while from the left aortic wall above the sinus in 6 (26.1%) patients only. Congenital heart disease and acquired valvular heart disease were the most common associated conditions. The author will share his experience and suggest a four-step approach of early recognition and selection of the anomalous right coronary artery ostium. Using the suggested strategy, most of the anomalous right coronary artery could be opacified with a left amplatz 1 catheter. Aortogram was needed only in 47.8% of cases. PMID- 15882156 TI - One stop shopping: providing today's competitive cardiovascular service. PMID- 15882157 TI - Use of stents to maintain atrial defects and fontan fenestrations in congenital heart disease. AB - Maintaining patent atrial septal communications or fenestrations can be vital in conditions requiring adequate decompression of the atria or Fontan baffle. We have recently deployed stents for this purpose, and the aim of this retrospective analysis is to describe our experience. All 26 patients undergoing such stent placement were retrospectively studied and for neonates with hypoplastic left heart syndrome (HLHS) and patients with Fontan fenestrations, their data were compared to controls undergoing transseptal static balloon dilation during the same time period. All 7 stented neonates with HLHS survived to their Norwood procedure and 57% survived to hospital discharge, similar to those who had static balloon dilation. Complications occurred in both HLHS groups but transient complete heart block was only seen in the control group, which also had larger balloons used (10.3 mm vs 7 mm, P=0.002). The success rate for patients undergoing stent placement in Fontan fenestrations was 64% compared to 76% with dilation alone. Complications were seen in 64% of the Fontan stented group compared to 39% for controls. There were 5 other patients with complex lesions (3 of whom were on the Extracorporeal Membrane Oxygenator) in whom stent placement successfully maintained atrial communication patency. Atrial septal stent placement in neonates with HLHS with restrictive defects is effective and appears at least as safe as static balloon dilation. On the other hand, initial fenestration stent placement is indicated only after extracardiac Fontan procedures in which the previous fenestration location cannot be found. PMID- 15882158 TI - "Taking in the waist": adjusting the size of a stented fontan fenestration. AB - Early spontaneous closure of a fenestration following Fontan palliation may complicate the postoperative management of such patients. The creation of a fenestration in the catheterization laboratory with an intravascular stent may improve these patients' hemodynamic status. The aim of this study is to present a new technique to reduce the diameter of stented Fontan fenestrations in those patients in whom the stent diameter is functionally made too large. PMID- 15882159 TI - Ingenuity in the catheterization laboratory. PMID- 15882160 TI - Primary angioplasty in a patient with dextrocardia. AB - Dextrocardia is a rare anomaly affecting 0.01% of newborns. We report on a case of a 60-year-old female with mirror-image dextrocardia who presented with an acute myocardial infarction and underwent primary angioplasty of the left circumflex artery. A review of the literature and technical challenges of the case will be addressed. PMID- 15882161 TI - Coronary artery stent thrombosis associated with heparin-induced thrombocytopenia: case report and review of the literature. AB - The diagnosis of an acute coronary syndrome associated with heparin-induced thrombocytopenia is an increasingly recognized complication of heparin exposure. We describe a case of an acute myocardial infarction 2 weeks after elective aortobifemoral bypass surgery, subsequent subacute coronary artery stent thrombosis, and the treatment strategies available. PMID- 15882162 TI - Acute left main occlusion during percutaneous coronary intervention associated with heparin induced thrombocytopenia with thrombosis syndrome. AB - A 76-year-old male was admitted with Braunwald IIIB unstable angina and treated with intravenous heparin. Coronary angiography 20 days later revealed a severe stenosis in the left circumflex artery. During coronary angioplasty thrombus developed in the circumflex artery, extended in the left main and lead to its occlusion. Normal left coronary artery patency and flow were achieved after intracoronary and intravenous administration of abciximab, and multiple stenting. Platelet-count decrease and an ELISA assay documented the presence of heparin induced thrombocytopenia with thrombosis syndrome (HITTS). HITTS should be suspected after acute thrombus formation during coronary angioplasty. PMID- 15882163 TI - The role of routine follow-up after gynecological malignancy. AB - The objective of this article was to determine the evidence base for routine follow-up after gynecological malignancy. Only articles with a survival analysis were included. Relevant articles were identified by a comprehensive literature search of the main biomedical databases, hand searching of references of selected articles, and expert spotting of relevant journals and proceedings of international meetings. A two-stage extraction of data was undertaken. No prospective trials were identified. Twenty-nine retrospective case series analyses and one poster presentation met the inclusion criteria. Eight articles and one letter on endometrial cancer, six articles and one poster presentation on cervical cancer, and two articles in vulval cancer were reviewed. Only one article in endometrial cancer showed any survival benefit from routine follow-up, but it was of very poor methodologic quality. Two articles found a survival benefit from routine follow-up after cervical cancer. The two articles on vulval cancer did not find any survival benefit from routine review. There is no prospective research on the benefits of routine follow-up after gynecological cancer. Retrospective evidence calls in to question the benefit of universal follow-up. Prospective research is urgently needed. PMID- 15882164 TI - Hormone replacement therapy on ovarian and uterine cancer risk and cancer survivors: how shall we do no harm? AB - This article will focus on the results of the Women's Health Initiative on the effects of hormone replacement therapy on coronary heart disease, stroke, venous thromboembolism, breast cancer, and colorectal cancer. Data from other relevant trials, including the most recent data on ovarian and uterine cancer risk and on gynecologic cancer patients, are also discussed to provide some guidelines on prescribing hormone replacement therapy in clinical practice, particularly in gynecologic cancer survivors. PMID- 15882165 TI - Preliminary toxicity analysis of intraperitoneal carboplatin in combination with intravenous paclitaxel chemotherapy for patients with carcinoma of the ovary, peritoneum, or fallopian tube. AB - The objective of this study was to provide preliminary toxicity data of multiple cycle combination chemotherapy with intraperitoneal (IP) carboplatin and intravenous (IV) paclitaxel for further clinical trials. The toxicity data of 42 patients with mullerian carcinoma who underwent IP carboplatin therapy in combination with IV paclitaxel were retrospectively analyzed. Chemotherapy was repeated through the Bard IP port placed at initial surgery using IV paclitaxel at 175 mg/m2 followed by IP carboplatin. The doses of carboplatin were either at area under the curve (AUC) = 5, 6, 6.5, 7, or 7.5. The toxicity data in a total of 237 cycles were analyzed. The median number of cycles for IP chemotherapy was 6 (range: 3-12). The incidences of maximal grade toxicities in all cycles were: grade (G)2/3 nausea/vomiting, 23.8%; G2/3 constipation, 42.9%; G2 abdominal pain, 28.6%; G2/3 sensory neuropathy, 14.3%; motor neuropathy, 4.8%; myalgia/arthralgia 33.4%; G3/4 neutrocytopenia, 85.4%; and G3/4 anemia, 35.4%. These were not related to the dose of carboplatin. The incidences of G3 thrombocytopenia in relation to the dose of carboplatin were AUC = 5, 0%; 6, 31.6%; 6.5, 44.4%; 7, 25.0%; and 7.5, 80%. G4 thrombocytopenia did not occur. A dose of carboplatin between AUC = 6 and 7 with IV paclitaxel at 175 mg/m2 is warranted for further evaluation. PMID- 15882166 TI - Clear-cell cancer of the ovary-is it chemosensitive? AB - The records of all patients with clear-cell ovarian cancer (CCC) who underwent complete surgical staging and chemotherapy between 1984 and 2001 were reviewed and 39 patients identified as suitable for study. The mean patient age was 56 years, and the stage distribution was as follows: stage I, 53%; stage II, 13%; stage III, 32%; and stage IV, 2%. One in three patients with stage I disease developed recurrent disease despite adjuvant chemotherapy. Seventy percent of tumors demonstrated a response to combination carboplatin and paclitaxel. Tumors which had either a partial response or failed to respond to first-line chemotherapy demonstrated no response to second-line nonplatinum chemotherapy. Endometriosis was identified in 31% of tumors, and 18% of patients developed deep venous thrombosis (DVT); however, neither endometriosis nor DVT was associated with a poorer outcome. CCC has a high recurrence rate in early-stage disease despite adjuvant treatment with cytotoxic chemotherapy. Advanced disease does respond to carboplatin and paclitaxel, which should be the chemotherapeutic regimen of choice. New second-line agents are urgently required. PMID- 15882167 TI - Quantitative biochemical analysis of lactate dehydrogenase in human ovarian tissues: correlation with tumor grade. AB - In an attempt to identify glycolytic capacity of normal and neoplastic human ovary, total lactate dehydrogenase (LDH) activity was measured in tissue cytosol originating from 69 patients (18 with benign ovarian tumor, 34 with ovarian carcinoma, six with nonepithelial ovarian malignant tumors, and 11 with tumor metastatic to ovary) and compared to the LDH activity of normal ovarian tissues (n = 19). Median value of total LDH-specific activity expressed as U/mg protein was 0.546 in normal tissues, 0.584 in benign tumors, 1.071 in malignancies metastatic to ovaries, 0.872 in nonepithelial primary ovarian tumors, and 0.818 in primary carcinomas. A significant rise in LDH-specific activity was found in malignant primary and secondary tumors of epithelial and nonepithelial origin, but not in benign neoplasms, compared to the activity in normal tissue. Ovarian carcinomas of serous histologic type did not differ in LDH activity from mucinous tumors. However, poorly differentiated carcinomas (grade 3) showed significantly enhanced activity of this glycolytic enzyme when compared to its grade 1 counterpart. The subgroup of grade 1 tumors did not differ in LDH activity from normal and benign ovarian tissue. Obtained results suggest that direct correlation might exist between ovarian epithelial tumor grade and lactate dehydrogenase activity. PMID- 15882168 TI - Chemosensitivity testing of ovarian cancer using the histoculture drug response assay: sensitivity to cisplatin and clinical response. AB - Despite cytoreductive surgery and chemotherapy, the prognosis of advanced ovarian cancer is still poor. Predicting the chemosensitivity of tumors might improve the outcome. Therefore, we investigated the clinical value of the histoculture drug response assay for ovarian cancer. Tumor specimens were cultured for 7 days on collagen gel sponge in medium containing cisplatin, and the 50% inhibitory concentration was determined by the 3-(4,5-dimethyl-2-thiazolyl)-2,5 diphenyltetrazolium bromide assay. Then the in vitro sensitivity to cisplatin was compared with the clinical response and survival. Apoptosis of tumor cells was also investigated. Among 173 ovarian cancer patients, 164 were evaluable by the assay, and 29 patients had measurable lesions for which the clinical response could be determined. The 5-year survival rate was significantly higher in patients with chemosensitive tumors than in those with chemoresistant tumors when the cutoff value was set at a 50% inhibitory concentration of 25 microg/mL and the accuracy of the assay was 82.8% (24/29). As chemosensitivity to cisplatin became greater, the number of apoptotic cells also increased. This chemosensitivity assay may help predict the clinical response to cisplatin-based chemotherapy, thus improving the survival of ovarian cancer patients. PMID- 15882169 TI - Diagnostic approach using the expression profiling of the P53 tumor suppressor gene and its related proteins in ovarian epithelial tumors. AB - The initial aim of this study was to examine the expression profiles of P53 and its upstream genes, downstream genes, and cell cycle regulators to determine whether these markers are useful for making a differential diagnosis among the benign, borderline, and malignant ovarian epithelial tumors. Between borderline and malignant tumors, the increased expression levels of P53, Bax, Cyclin E, and cyclin-dependent kinase-2 as well as the decreased expression levels of growth arrest and DNA damage (GADD45) and murine double minute-2 (MDM2) were significantly associated with malignancy (P < 0.01, each). Using the receiver operating curve (ROC), the most reliable cutoff value of the added-up staining scores of those markers was 4.5 with 79% sensitivity and 89% specificity for malignancy. Between benign and borderline tumors, the P21 and Bax expression levels were significantly higher in borderline tumors, whereas the Bcl-2 expression level was much higher in benign tumors (P < 0.01, each). Using the ROC, the cutoff value of the added-up staining scores used to discriminate between the two groups was 2.5 with 70% sensitivity and 74% specificity for borderline tumors. Thus, for the differential diagnosis between borderline and malignant tumors, the cutoff value 4.5 of the cumulative staining scores can be used. However, the cutoff value 2.5 for discrimination between benign and borderline tumors may not be useful because of its relatively low sensitivity and specificity. In addition, the P53, GADD45, Cyclin E, and MDM2 expression levels in malignant ovarian tumors might be useful for determining the histologic grade and type. PMID- 15882170 TI - Ultrastructure alterations in adriamycin-resistant and cisplatin-resistant human ovarian cancer cell lines exposed to nonlethal ultrasound. AB - Human ovarian cancer cell lines, SKOV3 and its adriamycin-resistant substrain SKOV3/ADR and COC1 and its cisplatin-resistant substrain COC1/DDP, were exposed to nonlethal ultrasound. Ultrastructures in sham-insonated and insonated cells were inspected by transmission electron microscopy, and cytochrome C in cytosol was determined by enzyme-linked immunosorbent assay. Ultrasound exposure led to no significant changes in SKOV3/ADR cells, but tumid mitochondria occurred in SKOV3 cells. Mitochondria changes were also detected in some exposed COC1 and COC1/DDP cells. Apoptotic bodies could be detected in either control or insonated COC1/DDP cells. A few exposed COC1/DDP cells became reticular. Cytochrome C in cytosol in exposed SKOV3/ADR cells was increased but that in exposed COC1/DDP cells was decreased. These findings revealed that the bioeffect of ultrasound on chemosensitive cells was not identical to that of chemoresistant ones, and ultrasound was a potential approach for treatment of drug-resistant ovarian cancers. PMID- 15882171 TI - Effect of radiotherapy on pelvic lymph node metastasis in cervical cancer stages IB2-IVA: a retrospective analysis of two comparative series. AB - Efficiency of radiotherapy in controlling lymph node metastasis is a controversial issue. A continuous series of 87 patients affected by cervical cancer stages IB2-IVA and treated using pelvic radiotherapy is presented. A retrospective comparison is made between two populations. In the two populations, a staging lymphadenectomy was carried out before the onset of the therapeutic program. In the first population (53 patients), the pelvic nodes only were dissected and in the second one (34 patients), the pelvic lymph nodes were left in place and the paraaortic nodes only were dissected. In both series, a completion surgery was performed after finalization of the radiotherapy. It was carried out at open abdomen in both series. It included a systematic pelvic dissection for the patients whose pelvic nodes had been intentionally left in place at the time of the initial staging lymphadenectomy. Both series were identical as far as classic risk factors were concerned (FIGO stage, maximal tumor diameter, lymphovascular space involvement). The radiotherapy administered to the pelvis was the same in both populations. The number of patients with pelvic lymph node metastasis was 21 (39.6%) in the first population versus 6 (17.6%) in the second one (P = 0.03). The percentage of positive lymph nodes among the retrieved lymph nodes was 18.94 in the first population versus 2.8 in the second one (P = 0.0001). Pelvic radiotherapy is likely to control most of the pelvic lymph node metastasis, but not all of them. Practical deductions and further developments are discussed. PMID- 15882172 TI - Pelvic exenterations for gynecological malignancies: twenty-year experience at Roswell Park Cancer Institute. AB - The objective of this study is to review the experience with pelvic exenterations for gynecological malignancies at our cancer institute. Charts of 48 women who underwent a pelvic exenteration between January 1980 and December 1999 were reviewed, and several outcomes were analyzed. Majority of patients had received prior radiation therapy. The median survival was 35 months, and the disease-free survival was 32 months. Mortality from the procedure was 4.2%. Early and late postoperative complication rates were 27% and 75%, respectively. Recurrence rate was 60%. Eight patients received intraoperative radiation. Median survival in this group was 11.3 vs 35 months (P = 0.003). Univariate analysis failed to show an association between type of pelvic exenteration, type of fecal and urinary diversion, outcome, need for reoperation, and recurrence. Contemporary pelvic exenterations are associated with a low mortality and a potential for long-term survival in a subset of patients who historically have been given a poor prognosis. In patients with recurrent gynecological cancer confined centrally to the pelvis, pelvic exenteration still remains the choice of therapy as response to chemotherapy to a centrally recurrent tumor in radiated area continues to be poor. Intraoperative radiation in select few patients needs to be further studied. PMID- 15882173 TI - Comparative study of chemoradiation and neoadjuvant chemotherapy effects before radical hysterectomy in stage IB-IIB bulky cervical cancer and with tumor diameter greater than 4 cm. AB - Tumor size seems to be a determinant in the prognosis of early cervical cancer. Patients with tumor greater than 4 cm (bulky) in diameter have worse outcome. The purpose of this study was to compare the efficacy of preoperative combined chemoradiation and neoadjuvant chemotherapy (NAIC) programs followed by radical hysterectomy in stage IB-IIB bulky cervical cancer. From September 1999 to April 2002, 60 patients with stage IB-IIB bulky cervical cancer were treated with preoperative external-beam radiotherapy to 45 Gy plus weekly cisplatin 50 mg/m2 or preoperative NAIC by cisplatin 50 mg/m2 and vincristin 1 mg/m2 every 7-10 days, for three courses. Surgery was performed 4-6 weeks after the completion of the preoperative treatment. There were no significant difference between age, stage, tumor size, and histopathologic type in two groups (P > 0.05). Toxicity associated with two treatment methods was usually mild. In chemoradiation group, two patients developed vesicovaginal fistula, and four patients developed long term hydronephrosis that needed urethral stenting. Before surgery, complete and partial clinical response had no significant difference between two groups (P > 0.05). After surgery, lymph node and parametrial involvement had no significant difference between two groups (P > 0.05). In NAIC group, more patients had significantly residual tumor (P = 0.012), but residual tumor size had no significant difference between two groups (P > 0.05). Pathologic complete response was significantly higher in chemoradiation group (P = 0.004). According to the result of this study, it seems that NAIC and chemoradiation had similar effects in survival prognostic factors. PMID- 15882174 TI - Can we substitute brush cytology for biopsy in the evaluation of cervical lesions under the guidance of colposcopy? AB - In cervical cancer screening, colposcopically directed biopsy is the gold standard method for identifying intraepithelial and occult invasive lesions of the uterine cervix. As biopsy needs special expertise and the procedure is not convenient for the patients, we sought to evaluate colposcopically directed brush cytology as a substitute for biopsy of cervical lesions. We studied a series of 150 women who were referred for colposcopic evaluation. Colposcopically directed brush cytology and biopsy were performed for all patients with abnormal colposcopic findings. A total of 40 samples were excluded due to unsatisfactory report of brush cytology. Of the remaining 110 samples, 34 abnormal pathologies were reported in biopsy evaluations, while only 9 abnormal cytologies were reported in brush cytology specimens. Brush cytology sensitivity and specificity were 26% and 97%, respectively. We conclude that colposcopically directed brush cytology is not a safe substitute for biopsy in the evaluation of cervical lesions. PMID- 15882175 TI - Radioguided sentinel lymph node detection in vulvar cancer. AB - Lymph node status is the most important prognostic factor in vulvar cancer. Histologically, sentinel nodes may be representative of the status of the other regional nodes. Identification and histopathologic evaluation of sentinel nodes could then have a significant impact on clinical management and surgery. The aim of this study was to evaluate the feasibility and diagnostic accuracy of sentinel lymph node detection by preoperative lymphoscintigraphy with technetium-99 m labeled nanocolloid, followed by radioguided intraoperative detection. Nine patients with stage T1, N0, M0, and 11 patients with stage T2, N0, M0 squamous cell carcinoma of the vulva were included in the study. Only three cases had lesions exceeding 3.5 cm in diameter. Sentinel nodes were detected in 100% of cases. A total of 30 inguinofemoral lymphadenectomies were performed, with a mean of 10 surgically removed nodes. Histological examination revealed 17 true negative sentinel nodes, 2 true positive, and 1 false negative. In our case series, sentinel lymph node detection had a 95% diagnostic accuracy, with only one false negative. Based on literature evidence, the sentinel node procedure is feasible and reliable in vulvar cancer; however, the value of sentinel node dissection in the treatment of early-stage vulvar cancer still needs to be confirmed. PMID- 15882176 TI - Outcomes of abandoned radical hysterectomy in patients with stages IB-IIA cervical cancer found to have positive nodes during the operation. AB - The objective of this study was to evaluate the outcomes of stages IB-IIA cervical cancer patients whose radical hysterectomy (RH) was abandoned for positive pelvic nodes detected during the operation compared with those found to have positive nodes after the operation. Among 242 patients with planned RH and pelvic lymphadenectomy (RHPL) for stages IB-IIA cervical cancer, 23 (9.5%) had grossly positive nodes. RH was abandoned, and complete pelvic lymphadenectomy was performed. Of these 23 patients, 22 received adjuvant chemoradiation, and the remaining 1 received adjuvant radiation. Four patients with positive para-aortic nodes were additionally treated with extended-field irradiation. When compared with 35 patients whose positive nodes were detected after the operation, there were significant differences regarding number of positive nodes and number of patients receiving extended-field irradiation. Complications in both groups were not significantly different, but the 2-year disease-free survival was significantly lower in the abandoned RH group compared with that of the RHPL group (58.5% versus 93.5%, P= 0.01). In conclusion, the survival of stages IB-IIA cervical cancer patients whose RH was abandoned for grossly positive pelvic nodes was significantly worse than that of patients whose node metastasis was identified after the operation. This is because the abandoned RH group had worse prognostic factors. PMID- 15882177 TI - Upregulation of heat shock protein 27 in metaplastic and neoplastic lesions of the endocervix. AB - Heat shock proteins (hsps) are molecular chaperones that are known to play a pivotal role in regulating intracellular homeostasis. hsp27 may have diagnostic and prognostic values for different gynecological malignancies. A cross-sectional analytical study was conducted at the Department of Pathology, The University of Liverpool, Liverpool, UK. Included in the study were 80 cervical glandular lesions of various histologic types, representing tuboendometrial metaplasia/endometriosis (n = 19), cervical glandular intraepithelial neoplasia (n = 33), and invasive adenocarcinoma (n = 28). Paraffin-embedded sections were stained using a commercial mouse monoclonal anti-hsp27 antibody with prior pressure-cooking for antigen retrieval. Sections of 11 normal cervices were used as controls. The median percentage of cells expressing hsp27 in each group was calculated. Normal cervical glands showed minimal expression of hsp27 (median: 10%, interquartile ranges [IQ]: 5-15). Expression was significantly more widespread in tuboendometrial metaplasia/endometriosis (median: 35%, IQ: 15-80), cervical glandular intraepithelial neoplasia (median: 60%, IQ: 32-80), and invasive adenocarcinoma (median: 40%, IQ: 25-80) when compared with normal endocervix (P = 0.007, < 0.001, and 0.001, respectively). However, no significant difference in hsp27 protein expression was found between cervical glandular intraepithelial neoplasia and invasive adenocarcinoma. In invasive adenocarcinoma, hsp27 showed no correlation with tumor grade, lymph node involvement, and lymphovascular space invasion. Our data highlight early dysregulation of hsp27 expression in both metaplastic and neoplastic lesions of the cervix. PMID- 15882178 TI - Heterogeneity of stage IIIA endometrial carcinomas: implications for adjuvant therapy. AB - The purpose of this study was to evaluate overall survival (OS) and determine prognostic subclassifications for stage IIIA endometrial cancer. Stage IIIA endometrial cancer patients treated at M.D. Anderson Cancer Center from 1989 to 2002 were reviewed. Clinical information was obtained from the medical record. Cox regression analyses were performed to evaluate the association of pathologic criteria and OS. Patients were divided into four groups based on this analysis: E1, endometrioid/pelvic cytology only; E2, endometrioid/adnexa +/- serosal spread; NE1, nonendometrioid/pelvic cytology only; and NE2, nonendometrioid/adenexa +/- serosal spread. Forty-nine patients were identified. By multivariate analysis, histology and extent of disease were the only factors associated with OS. Five-year OS in the four subgroups based on histology and extent of disease were: E1, 79%, E2, 65%, NE1, 64%, and NE2, 13%. Histologic subtype and extent of pelvic disease are the only prognostic factors associated with OS. Patients with endometrioid tumors and extent of pelvic disease limited to positive cytology had a favorable outcome, with or without adjuvant therapy. Future prospective clinical trials should consider subclassifying patients with stage IIIA disease to better evaluate the role of adjuvant therapy. PMID- 15882179 TI - Abnormal cervical cytology: a risk factor for endometrial cancer recurrence. AB - The objective of this study was to evaluate the relationship between cervical cytology, histologic type, and risk of endometrial cancer recurrence. We performed a retrospective study of patients undergoing surgery for endometrial carcinoma. Risk factors for recurrence including histology, tumor grade, nodal status, myometrial invasion, peritoneal washings, stage, and cervical cytology were assessed. Abnormal cervical cytology was defined as the presence of any endometrial cells on Pap smear. Papillary serous and clear cell carcinomas were considered high-risk histologies. Univariate and multivariate analyses of risk factors for recurrence were performed. Thirty-nine (9%) patients developed recurrent endometrial cancer. More patients with abnormal Pap smears recurred (12% versus 4%, P < 0.05). For endometrioid adenocarcinoma, abnormal cervical cytology occurred in 61% and 7% recurred, while with high-risk histologies, 84% had abnormal cervical cytology and 19% recurred (P < 0.05). Other significant predictors of recurrence on univariate analysis were myometrial invasion, nodal status, washings, stage, and histology. On multivariate analysis, only nodal status remained a significant predictor of recurrence. Abnormal cervical cytology is associated with increased risk of endometrial cancer recurrence. Abnormal cervical cytology occurs more frequently in high-risk histologies, which are known to have a higher risk of recurrence. On multivariate analysis, only nodal spread remains a significant predictor of recurrence. PMID- 15882180 TI - Receptorial and mitochondrial apoptotic pathways in normal and neoplastic human endometrium. AB - Under normal conditions, in human endometrium, apoptotic and antiapoptotic factors play an important role in tissue homeostasis. Abnormalities of apoptosis, a process implicated in several events in the reproductive organs, may contribute to neoplastic transformation. The present study aimed to investigate the involvement of both the receptorial and the mitochondrial pathways of apoptosis in normal endometrium and in endometrial carcinoma, by measuring caspase-3 and caspase-8 activities and cytosolic cytochrome c levels. Twelve endometrial carcinomas and nine normal endometrial specimens (four in mild proliferative phase, five in late secretory phase) were included in this study. Cytosolic fractions, obtained by differential centrifugation of tissue homogenates, were analyzed for caspase-3 and caspase-8 activities, as well as for cytochrome c content. Caspase-8 activity in normal secretory phase endometrium was higher than that in the proliferative phase and in the endometrial carcinoma. Moreover, higher cytochrome c levels were detected in endometrial carcinoma with respect to normal secretive endometrium. No significant differences were found in caspase-3 activity between normal and pathologic endometrium. The results obtained suggest that in normal endometrium, apoptosis takes place through the activation of both receptorial and mitochondrial pathways. Defects in both these pathways may contribute to the development of endometrial carcinoma. PMID- 15882181 TI - Is aromatase cytochrome P450 involved in the pathogenesis of endometrioid endometrial cancer? AB - Prospectively, the relationship between androgen levels in the utero-ovarian circulation, aromatase activity in endometrial and body fat tissue, and the presence or absence of endometrioid endometrial cancer was studied in postmenopausal women. In 43 women with endometrioid endometrial cancer and 8 women with a benign gynecological condition, a hysterectomy with bilateral salpingo-oophorectomy was performed. Using tritium water-release assays, aromatase activities in endometrial and body fat tissue were determined and related to the steroid levels from the peripheral and the utero-ovarian venous circulation (estradiol, androstenedione, testosterone) and to the presence or absence of endometrial cancer. Significant aromatase activity was found in both benign and malignant endometrial tissue samples. Aromatase activity in samples of endometrial tissue and in samples of body fat did not correlate with steroid levels in peripheral or utero-ovarian venous blood. Aromatase activity in samples of benign or malignant endometrium did not differ. Remarkably, in four women with mainly poorly differentiated endometrial cancer, very high aromatase activity was found in endometrial tissue. It is likely that multiple pathogenetic pathways exist that eventually lead to the formation of endometrioid endometrial cancer. The local availability of androgens and the finding that aromatase activity is present in both endometrial cancer and benign endometrial tissue support the hypothesis that aromatase activity in the endometrium may play a role in malignant transformation by converting androgens into mitogenic estrogens in the endometrial tissue. PMID- 15882182 TI - Expression of estrogen receptors ERalpha and ERbeta in endometrial hyperplasia and adenocarcinoma. AB - We assessed the expression of estrogen receptors (ER)alpha and ERbeta in 114 human endometrial hyperplasia and adenocarcinomas. The aim of this study was to determine the expression of both ER isoforms in human endometrial tissue by immunohistochemistry. From normal proliferation to simple and complex hyperplasia, the expression of ERalpha was increased (P < 0.05), while ERbeta did not change much (P > 0.05). In atypia hyperplasia and adenocarcinoma, both ERalpha and ERbeta were decreased significantly (P < 0.05). Most endometrial adenocarcinomas expressed ERalpha, either alone or in combination with ERbeta, and the ERbeta/ERalpha ratio was decreased when compared to normal proliferation (P < 0.05). Also, we found that the expression of ERalpha and ERbeta has no relationship with the status of lymph node of adenocarcinoma (P > 0.05). Both ERalpha and ERbeta play an important role in endometrial hyperplasia and carcinomas, the levels of ERalpha and ERbeta appear be used as prognostic indicators. PMID- 15882183 TI - Rectus abdominis myocutaneous flaps for neovaginal reconstruction after radical pelvic surgery. AB - The objective of this article is to compare the flap-specific complications associated with vertical (VRAM) and transverse (TRAM) rectus abdominis myocutaneous flap vaginal reconstructions performed during radical pelvic procedures. A retrospective chart review was performed to identify all patients who underwent VRAM and TRAM neovaginal reconstructions performed on the Gynecologic Oncology Service at Duke University Medical Center. Flap-specific complications were compared between the two techniques. From 1988 to 2003, 14 VRAM and 18 TRAM flap neovaginal reconstructions were performed on 32 women during the course of 22 (68%) total pelvic exenterations, 8 (25%) partial exenterations, and 2 (6%) radical vulvovaginectomies. Twenty-eight (88%) patients had been previously treated with radiation therapy or concurrent chemoradiation. Associated procedures included continent urinary conduit in 21 (66%), rectosigmoid reanastomosis in 8 (25%), and intraoperative or postoperative sidewall radiation therapy in 7 (22%) of patients. Overall median survival was 14 months (range: 2-week postoperative death to 65 months), with two (6%) acute postoperative mortalities. Fifteen flap-specific complications occurred in 12 (38%) patients, with no significant differences in flap type. Abdominal wound complications included four (12%) superficial wound separations, while one (3%) patient had a fascial dehiscence associated with complex fistulas that contributed to her death, but no patient developed incisional hernia. One patient each developed > 50% flap loss after TRAM and < 50% flap loss after VRAM flap, respectively. Four (12%) patients developed vaginal stricture or stenosis, two (6%) required percutaneous drainage of pelvic abscess or hematoma, and two (6%) developed rectovaginal fistula. Univariate analysis revealed a trend for increasing flap loss with body mass index > 35 (P = 0.056, Fisher exact two tailed test), but there were no significant associations with other patient characteristics or flap-specific complications. Thirteen (62%) of 21 patients who survived >12 months reported coitus. Both VRAM and TRAM are reliable techniques for neovaginal reconstructions after radical pelvic surgery and have a similar distribution of flap-specific complications involving the donor and recipient sites. PMID- 15882184 TI - Carcinosarcoma-like mural nodule in intestinal-type mucinous ovarian of borderline malignancy: a case report. AB - Epithelial ovarian tumors of borderline malignancy are tumors with histologic features and biologic behavior between benign and frankly malignant epithelial ovarian neoplasms. To date, we cannot accurately predict the patients who are prone to an aggressive course of disease. Here, we present a 35-year-old patient with carcinosarcoma-like mural nodule in intestinal-type mucinous ovarian tumor of borderline malignancy. Foci of intraepithelial carcinoma (about 10%) without stromal invasion are also noted. Total hysterectomy, bilateral salpingo oophorectomy, appendectomy, and omentectomy were performed, and the frozen pathology during operation showed mucinous tumor of borderline malignancy of left ovary on April 18, 2002. The patient was followed at our outpatient department for 19 months after operation and was free of the disease without any adjuvant chemotherapy. It is difficult to determine whether intestinal-type borderline mucinous tumors with intraepithelial carcinoma are associated with a worse prognosis compared with those with epithelial atypia alone due to disparate results in the published literature. In contrast, most patients with mural nodules of anaplastic carcinoma have had a malignant, often rapid, course. However, too few cases of carcinosarcoma-like mural nodule in mucinous tumor have been published to warrant a conclusion regarding their prognosis. PMID- 15882185 TI - Non-Hodgkin's lymphoma presenting as advanced ovarian cancer--a case report and review of literature. AB - Non-Hodgkin's lymphoma involving the ovaries is unusual and may cause confusion for the clinician since its presentation might resemble other, much more frequent tumors. Malignant lymphoid cells may occur in the ovary either as a primary neoplasm or as a secondary manifestation of a disseminated occult or known disease. The most common presenting signs or symptoms of malignant lymphomas involving the ovaries are abdominal or pelvic pain or mass. We present here a unique case of non-Hodgkin's lymphoma involving the ovaries presenting as advanced ovarian cancer with a pelvic mass, pleural effusion, and marked elevation of CA-125. PMID- 15882186 TI - Long-term progression-free survival of invasive uterine cervical carcinoma infected with human immunodeficiency virus: a case report. AB - We report the case of a patient with invasive uterine cervical carcinoma, who is also infected with human immunodeficiency virus. This patient has had the longest progression-free survival of any with acquired immunodeficiency syndrome uterine cervical carcinoma. She was found to be human immunodeficiency virus positive in February 1996 and found to have uterine cervical carcinoma stage IB in July 1996. Shortly thereafter, she underwent radiation therapy. She died of renal and liver failure in January 1999. However, no residual tumor existed at that time. The longest progression-free survival in this case may be attributable to maintenance of the CD4 cell count from the onset of uterine cervical carcinoma to death, which meant the patient's immune system to the cancer cells worked. PMID- 15882187 TI - Intracranial metastases from adenocarcinoma of cervix: a case report. AB - Carcinoma of cervix usually spreads contiguously to the adjacent organs, by lymphatics to the pelvic and para-aortic lymph nodes and by hematogenous route to distant organs. The common sites of distant metastasis are lung, supraclavicular lymph nodes, liver, and bones. The brain is a rare site of metastasis. We report here a rare case of a 72-year-old woman with carcinoma of cervix, FIGO stage IIA, who developed metastases to the brain. PMID- 15882188 TI - Massive myxoid leiomyosarcoma of the uterus. AB - Uterine myxoid leiomyosarcoma is a very rare and aggressive variant of uterine sarcoma, of which only 26 cases have been described in the literature published in English. A 59-year-old woman, who had been menopausal for 5 years and had a huge abdominal mass underwent laparotomy; histopathology of the bulky tumor revealed the particular characteristics of a myxoid leiomyosarcoma. Immunohistochemically, Ki-67 contributed toward indicating a malignancy, whereas p53 and progesterone and estrogen receptors were negative. The level of serum Ca125 was high at diagnosis, within normal limits after the fifth cycle of chemotherapy, and subsequently increased again at recurrence. Sixteen months after surgery, the patient died of respiratory failure. Immunohistochemistry and Ca125 levels can aid the diagnosis, management, and prognosis of uterine myxoid leiomyosarcoma. PMID- 15882189 TI - Adenocarcinoma of mammary-like glands in the vulva successfully treated by weekly paclitaxel. AB - An 87-year-old was referred for gynecologic evaluation of a lesion involving the left labia majus noted 3 years earlier. Fine-needle aspiration cytology revealed clusters with an acinous structure or glandular formation. The tumor appeared as cell clusters with linear arrangements. Histologic examination showed the same morphologic findings as scirrhus type of primary breast carcinoma. Examinations of the breasts and axillary lymph nodes were normal. This disease was diagnosed as an adenocarcinoma arising in mammary-like glands of the vulva. Bone scan showed multiple foci in the sternum, costa, and vertebrae, consistent with metastatic disease. We administered five courses of weekly paclitaxel chemotherapy, which achieved a partial response. There were no severe adverse effects. In our case, the fine-needle aspiration cytology was a rapid and minimally invasive method of diagnosis, and the findings were extremely similar to those of the scirrhus type of primary breast carcinoma. Rapid and accurate diagnosis made with this technique might contribute to a good prognosis in the early-staged cases. Weekly paclitaxel chemotherapy may be one of the safe and effective treatments for this disease with distant metastases, even in extremely aged patients (over 80 years). PMID- 15882191 TI - Invasive pulmonary aspergillosis in chronic obstructive pulmonary disease: an emerging fungal pathogen. AB - Acute invasive pulmonary aspergillosis occurs predominantly in immunocompromised hosts, with increasing numbers of cases of invasive aspergillosis among patients with chronic obstructive pulmonary disease (COPD) being reported. Among 13 cases of invasive aspergillosis diagnosed in COPD patients admitted to the intensive care unit with acute respiratory distress, the only risk factor for invasive fungal infection was corticosteroid treatment. Invasive aspergillosis should be suspected in COPD patients receiving steroid treatment who have extensive pulmonary infiltrates. Survival depends on rapid diagnosis and early appropriate treatment. A decrease or interruption of steroid treatment should be considered as part of the overall therapeutic strategy. PMID- 15882192 TI - Understanding how Listeria monocytogenes targets and crosses host barriers. AB - Human listeriosis is caused by the Gram-positive bacterium Listeria monocytogenes. In humans, this pathogen has the ability to cross the intestinal, placental and blood-brain barriers, leading to gastroenteritis, maternofetal infections and meningoencephalitis, respectively. The entry of L. monocytogenes into cultured human epithelial cells is mediated by the interaction of an L. monocytogenes surface protein, internalin, with its human receptor, E-cadherin. The internalin-E-cadherin interaction is species-specific, and relies on the nature of a single amino-acid in the E-cadherin molecule, which is proline in permissive species such as humans, and glutamic acid in non-permissive species such as the mouse. In a transgenic mouse model that expresses human E-cadherin in enterocytes, internalin allows L. monocytogenes to cross the intestinal barrier. Epidemiological evidence also supports a role for internalin in human listeriosis, not only for crossing the intestinal barrier, but also for targeting and crossing the placental and blood-brain barriers. Consistent with these epidemiological data, infection with L. monocytogenes of trophoblastic cell lines, primary trophoblast cultures and human placental villous explants demonstrates that bacterial invasion of the syncytiotrophoblast barrier is mediated by the internalin-E-cadherin interaction, leading to histopathological lesions that mimic those seen in the placentas of women with listeriosis. Thus, the internalin-E-cadherin interaction that plays a key role in the crossing of the intestinal barrier in humans is also exploited by L. monocytogenes to target and cross the placental barrier. Further investigations are currently focusing on the molecular mechanisms by which L. monocytogenes targets and crosses the blood brain barrier. PMID- 15882193 TI - Disruption of the interactions between the subunits of herpesvirus DNA polymerases as a novel antiviral strategy. AB - Most biological processes depend on the co-ordinated formation of protein-protein interactions. Besides their importance for virus replication, several interactions between virus proteins have been proposed as attractive targets for antiviral drug discovery, as the exquisite specificity of such cognate interactions affords the possibility of interfering with them in a highly specific and effective manner. There is a considerable need for new drugs active against herpesviruses, since available agents, most of which target the polymerisation activity of the virus DNA polymerase, are limited by pharmacokinetic issues, toxicity and antiviral resistance. A potential novel target for anti-herpesvirus drugs is the interaction between the two subunits of the virus DNA polymerase. This review focuses on recent developments using peptides and small molecules to inhibit protein-protein interactions between herpesvirus DNA polymerase subunits. PMID- 15882194 TI - Predominance of staphylococcal cassette chromosome mec (SCCmec) type IV among methicillin-resistant Staphylococcus aureus (MRSA) in a Swedish county and presence of unknown SCCmec types with Panton-Valentine leukocidin genes. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is an established nosocomial pathogen, but has recently begun to appear in the community. The clones in the community may not have originated in the hospital setting, and are referred to as community-acquired MRSA (CA-MRSA). Resistance to methicillin is mediated by the gene mecA, which is carried by the mobile genetic element staphylococcal cassette chromosome mec (SCCmec). SCCmec typing (I-IV) of all clinical isolates of MRSA (n = 92) from 1987 to 2004 in Orebro County, Sweden, was performed by real-time LightCycler PCR to detect the essential genetic components mecA, mecR1, IS1272, ccrA and ccrB. Forty-one isolates harboured type IV SCCmec, of which ten could be classified further as subtype IVa, and 27 as subtype IVc. No isolates belonged to subtype IVb, but four isolates could not be subtyped, and may be examples of novel type IV SCCmec subtypes. Thirty-five MRSA isolates, assigned to six different pulsotypes by pulsed-field gel electrophoresis, did not belong to SCCmec types I-IV. The Panton-Valentine leukocidin (PVL) genes were identified in two of these pulsotypes. Only SCCmec type IV has been associated previously with the PVL toxin, but the results suggest that new PVL-positive clones with novel SCCmec types may be arising and disseminating in the community. PMID- 15882195 TI - Effectiveness of a hospital-wide selective screening programme for methicillin resistant Staphylococcus aureus (MRSA) carriers at hospital admission to prevent hospital-acquired MRSA infections. AB - Screening of potential MRSA-positive patients at hospital admission is recommended in German and international guidelines. This policy has been shown to be effective in reducing the frequency of nosocomial MRSA transmissions in the event of an outbreak, but the influence of screening on reducing hospital acquired MRSA infections in a hospital setting where MRSA is endemic is not yet well-documented. This study describes the effect of hospital-wide screening of defined risk groups in a 700-bed acute care hospital during a period of 19 months. In a cohort study with a 19-month control period, the frequencies of hospital-acquired MRSA infections were compared with and without screening. In the control period, there were 119 MRSA-positive patients, of whom 48 had a hospital-acquired MRSA infection. On the basis of this frequency, a predicted total of 73.2 hospital-acquired MRSA infections was calculated for the screening period, but only 52% of the expected number (38 hospital-acquired MRSA infections) were observed, i.e., 48% of the predicted number of hospital-acquired MRSA infections were prevented by the screening programme. The screening programme was performed with minimal effort and can therefore be recommended as an effective measure to help prevent hospital-acquired MRSA infections. PMID- 15882196 TI - Cost analysis of a hospital-wide selective screening programme for methicillin resistant Staphylococcus aureus (MRSA) carriers in the context of diagnosis related groups (DRG) payment. AB - The costs of a hospital-wide selective screening programme were analysed for a period of 19 months. During this time, 539 inpatients were screened, of whom 111 were MRSA-positive. Based on microbiological costs (staff and materials) and the costs of preventive contact isolation for 2 days until microbiological results were available (including material costs for medical consumable goods and the costs of additional nursing time), a total of 26,241.51 Euro was spent for the 539 patients screened. Based on cost units, the costs were 39.96 Euro for a patient found to be MRSA-negative and 82.33 Euro for a patient found to be MRSA positive. Under the prospective diagnosis related groups (DRG) payment system in Germany, the costs of a prolonged hospital stay resulting from a hospital acquired MRSA infection (HA-MRSA-I) are not reimbursed adequately by revenues, with a calculated average cost-revenue loss/patient with HA-MRSA-I of 5705.75 Euro. The screening programme was able to prevent 48% of predicted HA-MRSA-Is (35.2 patients with infection), thereby saving a predicted 200,782.73 Euro. After subtracting the screening costs, there was a net saving of 110,236.56 Euro annually. A sensitivity analysis of the break-even points for different screening frequencies and different MRSA incidence rates indicated that the screening programme became cost-effective at a low MRSA incidence rate, meaning that it can be recommended for most hospitals with an MRSA problem. PMID- 15882197 TI - Interference of Lactobacillus plantarum with Pseudomonas aeruginosa in vitro and in infected burns: the potential use of probiotics in wound treatment. AB - This study evaluated the ability of the probiotic organism Lactobacillus plantarum to inhibit the pathogenic activity of Pseudomonas aeruginosa, both in vitro and in vivo, and investigated the mechanisms involved in such protection. L. plantarum whole cultures, culture filtrates (acid filtrate and neutralised acid filtrate) and isolated, washed cells were tested in vitro for their effects on the production of the P. aeruginosa quorum-sensing signal molecules, acyl homoserine-lactones (AHLs), and two virulence factors controlled by these signal molecules, elastase and biofilm. All were inhibited by L. plantarum cultures and filtrates, but not by isolated, washed cells. The acid L. plantarum growth medium itself had some inhibitory activity, but the greatest activity was exerted by the whole culture. To test the in-vivo activity of L. plantarum, a burned-mouse model was used in which burns infected with P. aeruginosa were treated with L. plantarum at 3, 4, 5, 7 and 9 days post-infection. Samples from skin, liver and spleen taken after 5, 10 and 15 days demonstrated inhibition of P. aeruginosa colonisation by L. plantarum. There was also an improvement in tissue repair, enhanced phagocytosis of P. aeruginosa by tissue phagocytes, and a decrease in apoptosis at 10 days. These results indicate that L. plantarum and/or its by products are potential therapeutic agents for the local treatment of P. aeruginosa burn infections. PMID- 15882198 TI - Tc-99m polyclonal human immunoglobulin scintigraphy in brucellosis. AB - The aim of this study was to evaluate the accuracy of Tc-99m polyclonal human immunoglobulin (HIG) scintigraphy for the diagnosis of brucellosis, and to compare its effectiveness in the diagnosis of osteoarticular involvement in comparison with bone scanning. Of 30 patients with brucellosis, Tc-99m HIG detected osteoarticular involvement in 18 (60%) patients, in whom the sacroiliac joints were affected most commonly (n = 13; 72.2%), with statistically predominant bilateral involvement (p < 0.05). By bone scanning, the rate of osteoarticular involvement was 70% (21 of 30 patients), and the joints affected most commonly were sacroiliac (15 of 21 patients; 71.4%). Although bilateral involvement was observed mostly by bone scanning, there was no significant difference between the rate of bilateral and unilateral involvement. The anatomical distribution of osteoarticular complications, as detected by Tc-99m HIG and bone scintigraphy, did not differ significantly. With Tc-99m HIG, orchitis was detected in two patients and paravertebral abscess in one patient. Since bone scanning did not detect these soft tissue complications, Tc-99m HIG scintigraphy might be useful for the detection of both osteoarticular and soft tissue complications resulting from brucellosis. PMID- 15882199 TI - Correlation between the procedure for antifungal susceptibility testing for Candida spp. of the European Committee on Antibiotic Susceptibility Testing (EUCAST) and four commercial techniques. AB - The correlation between results obtained with the European Committee on Antibiotic Susceptibility Testing (EUCAST) antifungal susceptibility testing procedure (document 7.1) and four commercial systems was evaluated for a collection of 93 clinical isolates of Candida spp. Overall, agreement between the EUCAST procedure and the Sensititre YeastOne and Etest methods was 75% and 90.4%, respectively. The correlation indices (p < 0.01) between the EUCAST and commercial methods were 0.92 for Sensititre YeastOne, 0.89 for Etest, - 0.90 for Neo-Sensitabs, and 0.95 for Fungitest. Amphotericin B MICs obtained by Sensititre YeastOne were consistently higher than with the EUCAST method and, although very major errors were not observed, 91% of MICs were misclassified. Amphotericin B- and fluconazole-resistant isolates were identified correctly with Sensititre YeastOne, Etest and Fungitest. Neo-Sensitabs identified amphotericin B-resistant isolates, but misclassified > 5% of fluconazole-resistant isolates as susceptible. The commercial methods, particularly Etest and Fungitest, appeared to be suitable alternatives to the EUCAST procedure for antifungal susceptibility testing of clinical isolates of Candida. PMID- 15882200 TI - Diagnosis of Candida lung abscesses by 18F-fluorodeoxyglucose positron emission tomography. AB - In three patients with catheter-associated candidaemia, use of 18F fluorodeoxyglucose positron emission tomography (FDG-PET) led to the diagnosis of Candida lung abscesses, which was confirmed by computed tomography and a favourable response to antifungal therapy. It was concluded that FDG-PET is a promising new imaging technique that enables early identification of sites of disseminated candidiasis, and that this technique can be used in the evaluation of therapy. PMID- 15882201 TI - Nocardiosis in a teaching hospital in the Central Anatolia region of Turkey: treatment and outcome. AB - Predisposing factors, antimicrobial susceptibility patterns, treatment and outcome were analysed for nine consecutive patients with nocardiosis. Predisposing factors were identified in six (67%) of the nine patients. Clinical syndromes of nocardial infection were pulmonary infection (three patients), cerebral infection (five patients) and disseminated infection (one patient). The predominant (60%) species was Nocardia farcinica rather than the Nocardia asteroides complex. Treatment was started empirically, modified according to the antimicrobial susceptibility pattern, and then continued for 6-12 months. Overall mortality was 33%, with death being caused by the Nocardia infection in two cases. PMID- 15882202 TI - Hepatitis B virus (HBV) genotyping in Belgian patients with chronic HBV infection. AB - Several studies have demonstrated that pathogenic and therapeutic differences exist among hepatitis B virus (HBV) genotypes. Therefore, this study established the prevalence of different HBV genotypes in 128 Belgian patients with chronic HBV infection. The prevalences of genotypes A and D, and mixed genotypes A and D, were 53%, 37% and 8%, respectively, for a group of blood donors, and 54%, 31% and 9%, respectively, for a group of patients from the gastroenterology units. The results indicated that genotypes A and D are the predominant genotypes in Belgian patients with chronic HBV infection. PMID- 15882203 TI - Thirty-day mortality of nosocomial systemic bacterial infections according to antibiotic susceptibility in an 800-bed teaching hospital in France. AB - Overall 30-day mortality among patients with nosocomial bacterial infections was analysed according to antibiotic susceptibility in an 800-bed hospital. Survival analysis identified three factors associated with an increased mortality rate: (1) a longer interval between admission and onset of infection; (2) bacterial resistance; and (3) the severity of the initial illness in the intensive care unit. The increased mortality rate associated with antibiotic resistance was observed for all bacterial species with the exception of Gram-positive cocci, and remained significant or near-significant regardless of the hospital ward. PMID- 15882204 TI - Differences in the proportions of fluoroquinolone-resistant Gram-negative bacteria isolated from bacteraemic children with cancer in two Italian centres. AB - The proportion of ciprofloxacin-resistant Gram-negative bacteria isolated from the blood of children with cancer (not receiving prophylaxis) was 10% in a paediatric hospital (Genoa) where the use of quinolones was highly restricted, compared with 41% in a department of haematology (Rome) where leukaemic adults, who received fluoroquinolone prophylaxis, were also treated (p < 0.0001). Moreover, simultaneous resistance to ciprofloxacin and ceftazidime, amikacin or imipenem-cilastatin was 11% in Genoa compared with 37% in Rome (p < 0.001). Ciprofloxacin resistance was more frequent in children who shared an environment with adults who were receiving ciprofloxacin prophylaxis. PMID- 15882205 TI - Elevated serum alanine aminotransferase (ALT) levels in patients with serologically verified Mycoplasma pneumoniae pneumonia. AB - The possibility of liver involvement in Mycoplasma pneumoniae pneumonia is still controversial. This study investigated 33 adult patients with serologically confirmed M. pneumoniae community-acquired pneumonia (CAP) (median age 31 years) and 38 patients with bacteraemic Streptococcus pneumoniae CAP (median age 54 years), all without pre-existing liver disease. Serum alanine aminotransferase (ALT) levels were elevated in 12 (36.4%) patients with M. pneumoniae CAP (median 53.5 U/L), and in four (10.5%) patients with S. pneumoniae CAP (median 61 U/L) (p 0.025). In most patients with M. pneumoniae CAP, the elevated ALT levels decreased during macrolide therapy, although this decrease was not significant. PMID- 15882206 TI - Activity of medicinal plant extracts against hospital isolates of methicillin resistant Staphylococcus aureus. AB - Aqueous and ethanolic extracts of ten traditional Thai medicinal plants were investigated for their ability to inhibit 35 hospital isolates of methicillin resistant Staphylococcus aureus (MRSA). Nine medicinal plants displayed activity against all isolates tested. Ethanolic extracts of Garcinia mangostana, Punica granatum and Quercus infectoria were most effective, with MICs for MRSA isolates of 0.05-0.4, 0.2-0.4 and 0.2-0.4 mg/mL, respectively, and for S. aureus ATCC 25923 of 0.1, 0.2 and 0.1 mg/mL, respectively. MBCs for MRSA isolates were 0.1 0.4, 1.6-3.2 and 0.4-1.6 mg/mL, and for S. aureus ATCC 25923 were 0.4, 3.2 and 1.6 mg/mL, respectively. PMID- 15882208 TI - Influence of alcohol in mouthwashes containing triclosan and zinc: an experimental gingivitis study. AB - OBJECTIVES: An experimental gingivitis model was used to analyse the influence of alcohol in mouthwashes containing 0.15% triclosan and zinc chloride on the formation of supragingival plaque, the development of gingivitis and the appearance of adverse events. MATERIAL AND METHODS: Using a double-blind crossover design, 30 subjects underwent two consecutive experimental phases with two 0.15% triclosan and zinc chloride mouthwashes, differentiated mainly by their excipient (hydroalcoholic or aqueous). In each phase, the subjects discontinued all oral hygiene measures and were treated solely with the randomly assigned experimental mouthwash for 21 days. Each experimental phase was preceded by a 14 day washout period in which, after receiving a complete oral prophylaxis, the subjects were instructed to perform thorough oral hygiene procedures. Gingivitis and plaque levels were assessed at the start and end of both the experimental phases. RESULTS: The evolution of the gingivitis and plaque indices showed no statistically significant differences between both treatments. The mean gingival index increased from 0.30 to 0.38 with the non-alcoholic mouthwash and from 0.32 to 0.42 with the hydroalcoholic mouthwash. The respective changes in plaque index were from 0.88 to 1.93 and from 0.94 to 1.92. The hydroalcoholic mouthwash produced a larger number of fresh symptoms (p=0.033), oral itching being the most reported. CONCLUSIONS: Alcohol does not influence the effectiveness of a 0.15% triclosan and zinc chloride mouthwash against gingivitis development and supragingival plaque formation. The absence of alcohol significantly decreases the incidence of adverse events produced by the mouthwash. PMID- 15882209 TI - The prevention of plaque re-growth by toothpastes and solutions containing block copolymers with and without polypeptide. AB - BACKGROUND/AIMS: Chemicals which have a direct effect at inhibiting or reducing bacterial adherence to tooth surfaces may subsequently inhibit plaque growth and reduce gingival inflammation. This study investigated whether two anti-adherent systems could inhibit plaque re-growth in the mouth when rinsed as a solution or as a toothpaste slurry. METHODS: A total of 21 subjects took part in a partially blind, seven cell cross-over study which compared the effects on plaque re-growth of a binary system containing block copolymers, a ternary system containing block copolymers and polypeptide, both used as toothpaste slurry rinses, their corresponding solution rinses, a conventional fluoride toothpaste rinse, a positive control chlorhexidine rinse and a negative water control. Following a dental prophylaxis subjects then rinsed with 10 ml of one of the test products for 1 min. twice a day over a 4-day period. Throughout each trial period the subjects were not permitted to use any other forms of oral hygiene. On the fifth day (96 h), the volunteers returned to the clinic, and plaque was assessed by (1) plaque index and (2) plaque area following disclosing with a food dye. The test phase of the trial was repeated for each agent and was followed by a 10-day "washout" period. RESULTS: Essentially neither of the anti-adherent systems inhibited plaque re-growth, whether administered in a toothpaste slurry or solution compared with the controls. If anything, neither of the test pastes were as effective as the marketed commercial paste (p<0.001). As expected plaque recorded following use of the chlorhexidine rinse was significantly less than that seen with any of the other rinses (p<0.001). CONCLUSIONS: Using this 4-day plaque re-growth model, the findings of this study failed to show any benefit in using the anti-adherent systems, either in a rinse or toothpaste, with the aim of inhibiting or reducing plaque formation. PMID- 15882210 TI - Tobacco smoking and periodontal microflora in a Saudi Arabian population. AB - OBJECTIVES: To explore the subgingival periodontal microflora in a Saudi Arabian population with a special focus on its relationship with various smoking habits. MATERIAL AND METHODS: A total of 198 individuals in the age range 17-60 years were included in the study. 29% were water-pipe smokers, 18% cigarette smokers, 13% smokers of both water pipe and cigarettes (mixed smokers) and 40% non smokers. For each individual, a subgingival plaque sample from the deepest site in each quadrant was obtained. The checkerboard DNA-DNA hybridization technology was used to determine the presence of Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Tannerella forsythensis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Treponoma denticola, Peptostreptococcus micros, Campylobacter rectus, Eikenella corrodens, Selenomonas noxia and Streptococcus intermedius. Two cut-off levels for detection were used, score 1(10(5) bacteria) and score 3 (10(6) bacteria). RESULTS: The prevalence of individuals positive for the different microorganisms at score 1 cut-off varied from 7% to 95%. At score 3 cut-off the prevalence varied from 0% to 30%. The depth of sample site was a key factor for detection. When the depth of sample site was taken into account, no statistically significant differences were observed between cigarette smokers, water-pipe smokers, and non-smokers with regard to occurrence of the microorganisms studied. CONCLUSIONS: No major differences were observed between cigarette smokers, water-pipe smokers, and non smokers regarding the occurrence of the periodontal microorganisms studied suggesting that this portion of the subgingival periodontal microflora is independent of tobacco smoking. PMID- 15882211 TI - Effect of periodic preventive care on the progression of periodontal disease in young adults with Down's syndrome. AB - OBJECTIVES: To examine the effect of periodic preventive care on the progression of periodontal disease in 24 young adults with Down's syndrome (DS) (mean age+/ SD: 20.8+/-5.6 years) MATERIAL AND METHODS: Subjects were divided into two groups: 13 subjects who had frequently visited our clinic (mean interval between visits: 3.7+/-1.3 months; managed group) and 11 subjects who had not visited our clinic for more than 1 year (mean duration of no visits: 27.5+/-10.1 months; interrupted group). The progression of periodontal disease in the subjects was evaluated clinically, microbiologically and roentgenologically. RESULTS: Clinical parameters (probing depth, frequency of the presence of a pathological periodontal pocket, alveolar bone loss, frequency of the incidence of pathological bone loss, and modified total PMA (M-PMA)) and benzoyl-DL-arginine naphthylamide (BANA) scores in the interrupted group were significantly higher than those in the managed group. Subject age showed significant positive correlations with probing depth and alveolar bone loss. BANA scores showed significant positive correlations with probing depth, alveolar bone loss and M PMA in the interrupted group. CONCLUSIONS: The results suggest that periodic preventive care is effective for suppressing the progression of periodontal disease in young adults with DS. PMID- 15882212 TI - Fibrin clot adhesion to dentin conditioned with protein constructs: an in vitro proof-of-principle study. AB - OBJECTIVES: Periodontal regeneration is contingent on the adsorption, uninterrupted adhesion, and maturation of a fibrin clot to a periodontally compromised root surface. Clot adhesion appears vitally dependent on the formation of a resilient union between the clot and the root surface. Root surface demineralization will remove a root surface smear layer exposing dentin tubules and collagen matrix for enhanced clot adhesion. Recently, protein constructs have been introduced to condition the root during periodontal surgery. The effect of such root conditioning on clot adhesion has not been clarified. The objective of this study was to evaluate clot adhesion to protein conditioned dentin surfaces. METHODS: Human dentin blocks (4 x 6 x 1 mm) were exposed to a saturated citric acid solution (CA) or a commercial ethylenediaminetetraacetic acid (EDTA) preparation using standardized protocols. Some dentin blocks were additionally conditioned with proteins, either bovine serum albumin (BSA) or an enamel matrix protein preparation (EMP). Fresh human whole blood was applied to the blocks. The blood was allowed to clot for 20 min. in a humidified chamber. The dentin blocks were rinsed 3 x 5 min. in phosphate-buffered saline under standardized conditions to test clot adhesion. They were then processed for scanning electron microscopy (SEM). Two masked examiners independently evaluated the SEM images. RESULTS: CA removed the dentin smear layer, exposing dentin tubules and collagen. EDTA appeared less efficacious leaving smear layer residues. The BSA or EMP application resulted in a surface morphology similar to that of a smear layer. Fibrin clot adhesion was best supported by the CA-treated dentin surface. Forces produced by the rinse protocol partially removed the fibrin clot from EDTA-treated surfaces. BSA- or EMP-treated surfaces poorly retained the fibrin clot. CONCLUSIONS: CA surface demineralization removes a dentin surface smear layer to promote adhesion of a fibrin clot. The EDTA gel appears less effective. Further conditioning of the dentin surface with protein constructs produces a surface morphology similar to that of the smear layer with poor fibrin clot retention. PMID- 15882213 TI - Implant-supported single-tooth restorations: a 5-year prospective study. AB - BACKGROUND: Comparatively few studies are available reporting at least 5 years of follow-up data of implant-supported single-tooth replacements. OBJECTIVE: To evaluate prospectively the 5-year outcome of implant-supported single-tooth prosthetic restorations. MATERIAL AND METHODS: Forty subjects (mean age 41 years), 23 males and 17 females, who required single-tooth prosthetic replacement for a missing tooth were recruited. A total of 45 self-tapping implants (Astra Tech ST-implants)--40 in the maxilla and five in the mandible--were installed in a two-stage procedure. Abutment connection was performed 3-6 months after implant installation. Clinical and radiographic examinations were performed at the completion of the prosthetic treatment and once a year during a 5-year follow-up period. The analysis of peri-implant bone level alteration was performed on subject and implant levels and by the use of analysis of variance and binary logistic regression. RESULTS: Three patients were lost during the 5 years of follow-up. One implant was lost after 2.5 years in function and another four implants could not be accounted for at the 5-year follow-up examination. The overall failure rate at 5 years was 2.6% (subject level) and 2.3% (implant level). The mean loss of marginal bone at the implants during the first year in function was 0.06 mm (SD 0.67) on the subject level and 0.02 mm (0.65) on the implant level. During the subsequent 4 years the annual change in peri-implant bone level amounted to -0.02 mm (0.22) on both subject and implant levels. Thus, the mean total bone level change over the 5-year interval was -0.14 mm (1.04) on subject level and -0.11 mm (1.00) on the implant level of analysis (p>0.05). The frequency of implants with a 5-year bone loss of > or =1 mm was 13%. Approximately 50% of the implants demonstrated no bone loss. CONCLUSION: The present clinical trial on single-tooth replacements with the Astra Tech implant system demonstrated that the bone loss during the first year of function as well as annually thereafter was small. PMID- 15882214 TI - Approximal plaque and gingival sulcus bleeding in routine dental care patients: relations to life stress, somatization and depression. AB - OBJECTIVES: Previous research on relations between life stress and periodontal conditions was often based on indicators of periodontal destruction history and stimulus-specific measures of life stress. The present study was undertaken to compare patients showing different degrees of gingival sulcus bleeding with patients without signs of bleeding, using an instrument measuring cognitive impacts of events and assessing reported symptoms of depression and somatization. Additionally the patient groups were compared with regard to dental status and dental visiting behaviour. Analog comparisons were conducted in patients with different degrees of approximal plaque. MATERIALS AND METHODS: 140 routine dental care patients attending a private dental practice were examined. Measurement devices were the sulcus bleeding index (SBI), the approximal plaque index (API), the Dental Avoidance Scale, the Life Experience Survey (LES), the Impact of Event Scale with the subtests cognitive intrusion and cognitive avoidance, and the SCL 90-R subscales somatization and depression. RESULTS: Patients with high API scores differed from their zero-score counterparts in dental avoidance and time since last visit only. By contrast, subjects scoring high in the SBI not only had more missing teeth and reported more dental avoidance than their zero-score counterparts but also had elevated scores in intrusive and avoidant thoughts related to a stressful event as well as in symptoms of somatization and depression. No difference was found for the measure of stimulus specific recent life events (LES). CONCLUSIONS: The results suggest that cognitive preoccupation with adverse events and the presence of somatization and depression symptoms might play a role in gingival inflammation. PMID- 15882215 TI - Periodontal healing in one-wall intra-bony defects in dogs following implantation of autogenous bone or a coral-derived biomaterial. AB - AIM: Autogenous bone grafts and bone biomaterials are being used as part of protocols aiming at reconstruction of periodontal defects. There is a limited biologic information on the effect of such materials on periodontal healing, in particular aberrant healing events that may prevent their general use. The objective of this study was, using histological techniques, to evaluate periodontal healing with focus on root resorption and ankylosis following implantation of autogenous bone and a coral-derived biomaterial into intra-bony defects in dogs. METHODS: One-wall intra-bony periodontal defects were surgically created at the distal aspect of the second and the mesial aspect of the fourth mandibular premolars in either right or left jaw quadrants in four Beagle dogs. Each animal received particulated autogenous bone and the resorbable calcium carbonate biomaterial into discrete one-wall intra-bony defects. The mucoperiosteal flaps were positioned and sutured to their pre-surgery position. The animals were euthanized 8 weeks post-surgery when block sections of the defect sites were collected and prepared for qualitative histological analysis. RESULTS: There were no significant differences in periodontal healing between sites receiving autograft bone and the coral-derived biomaterial. A well organized periodontal ligament bridging new bone and cementum regeneration was observed extending coronal to a notch prepared to delineate the apical extent of the defect. Osteoid and bone with enclosed osteocytes were formed onto the surface of both autograft and coral particles. Although small resorption pits were evident in most teeth, importantly none of the biomaterials provoked marked root resorption. Ankylosis was not observed. CONCLUSION: Particulated autogenous bone and the coral-derived biomaterial may be implanted into periodontal defects without significant healing aberrations such as root resorption and ankylosis. The histopathological evaluation suggests that the autogenous bone graft has a limited osteogenic potential as demonstrated in this study model. PMID- 15882216 TI - Effectiveness of a transmucosal lidocaine delivery system for local anaesthesia during scaling and root planing. AB - OBJECTIVE: This study compared the efficacy of transmucosal anaesthetic patches containing lidocaine (46.1 mg/2 cm(2)) to placebo for local anaesthesia during quadrant scaling and root planing using periodontal clinical indices and patient perception of pain. MATERIAL AND METHODS: Forty healthy adults with moderate periodontal disease and moderate subgingival calculus were scaled at weekly intervals, two quadrants randomized to treatment patches and two quadrants randomized to placebo patches. Bleeding, probing depths and attachment levels were evaluated prior to treatment and 1 month after quadrant scaling was completed. Subjects completed 100 mm visual analogue pain scales 15 min. after patch placement and at the end of treatment, and were asked for verbal assessment of perceived pain. RESULTS: Subjects' verbal ratings demonstrated consistently greater pain relief with active patches than placebo (p<0.0001). Visual analogue scales demonstrated significantly greater pain relief with the treatment patches after 15 min. (p=0.0003) and at the end of treatment (p=0.0149). Efficacy of periodontal therapy was equivalent for treatment and control groups. No adverse events were observed; localized minimal gingival irritation was noted in three subjects. CONCLUSION: Transmucosal lidocaine patches provided sufficient anaesthesia for therapeutic quadrant scaling and root planing procedures. PMID- 15882217 TI - Efficacy of a 0.15% benzydamine hydrochloride and 0.05% cetylpyridinium chloride mouth rinse on 4-day de novo plaque formation. AB - OBJECTIVE: To evaluate the effect of a mouth-rinse formulation combining benzydamine hydrochloride and cetylpyridinium chloride (BNZ+CPC) in preventing de novo plaque formation, in comparison with CPC and placebo mouth rinses. PATIENTS AND METHODS: This was a controlled, observer-blind, cross-over study. In this model of plaque re-growth, subjects received a session of oral prophylaxis and were directed to withdraw oral hygiene measures for the next 4 days, using only the mouth rinse assigned. The outcome parameters were the plaque index (PlI) and gingival index (GI). In addition, microbiological evaluation of the subgingival microflora, by means of culture, was performed, as well as patient-based variables. Data analysis was carried out using anova for Latin-square design. RESULTS: The analysis of variance showed a significant statistical difference between the BNZ+CPC association and placebo (p<0.0001). No differences between CPC and placebo were detected considering multiple comparisons between treatments. The 90% confidence interval of the differences between BNZ+CPC and CPC showed no equivalence between treatments, being the PlI lower in the BNZ+CPC group. No significant difference between groups in GI was observed. Mean anaerobic colony-forming units (CFU) demonstrated a significant increase between visits in all groups (p<0.001) and differences among groups were not significant. Subjects treated with BNZ+CPC frequently reported "tingling mouth" and "numbness mouth". CONCLUSION: Within the limitations of the study model, the BNZ+CPC combination showed a statistically significant plaque-inhibitory capacity, as compared with the placebo mouth rinse, and an additive effect as compared with CPC. No relevant clinical or microbiological adverse effects were detected. PMID- 15882218 TI - Quality of bristle end-rounding on replaceable heads of powered toothbrushes. AB - OBJECTIVES: The aim of this study was to evaluate the geometry and the quality of bristle tip-rounding using 14 different heads from powered toothbrushes. MATERIAL AND METHODS: Six powered toothbrushes for children and eight for juveniles and adults were included. Five replaceable heads of each product were randomly selected. Of each sample, 35 bristles were used for examination. This resulted in 175 bristles from each product being evaluated. The quality of end-rounding was assessed by scanning electron microscopy at an original magnification x 80 in two categories of acceptable and five categories of unacceptable rounding according to Silverstone & Featherstone (1988). RESULTS: The portion of acceptable end rounding varied strongly between the products (18.9-94.3%). There were significant differences regarding the products for children (p<0.001) and for adults (p<0.001) with respect to end-rounding quality. Only one product achieved more than 90% and eight products had between 68% and 86% acceptable end-rounding. Two products for children and one for adults had less than 25% acceptable end rounding. CONCLUSION: A high standard of bristle tip-rounding is an important feature with respect to the safety of powered toothbrushes. Those products with a greater portion of unacceptably rounded bristles might cause more harm to oral soft tissues during use. The end-rounding quality of some of the products should be improved. PMID- 15882219 TI - Adjunctive subantimicrobial dose doxycycline in smokers and non-smokers with chronic periodontitis. AB - OBJECTIVES: Previous studies have demonstrated the clinical benefits of sub antimicrobial dose doxycycline (SDD) in the treatment of chronic periodontitis (CP). The aim of this study was to retrospectively evaluate the role of SDD as an adjunct to scaling and root planing (SRP) in the treatment of smokers and non smokers with CP. MATERIAL AND METHODS: A meta-analysis of two previously reported clinical studies was undertaken. Both were 9-month, double-blind, randomized, placebo-controlled, multi-centre clinical trials that investigated the efficacy of SDD (20 mg doxycycline twice daily) in combination with SRP in subjects with moderate-severe CP. 36.9% of the combined study population were smokers. Three hundred and ninety-two subjects were included in the meta-analysis, which evaluated per-subject mean changes in clinical attachment level (CAL) and probing depth (PD) from baseline and the total number of sites with attachment gains and PD reductions > or =2 and > or =3 mm from baseline in four subgroups: smokers/SDD; smokers/placebo; non-smokers/SDD; non-smokers/placebo. RESULTS: A hierarchical treatment response was observed, with non-smokers who received SDD demonstrating the greatest CAL gains and PD reductions. Smokers who received placebo demonstrated the smallest clinical improvements following treatment. Smokers who received SDD demonstrated an intermediate treatment response that was broadly equivalent to that seen in non-smokers who received placebo. In sites with baseline PD 4-6 mm, month 9 CAL gains were 19-45% better in non-smokers who received SDD compared with all other subgroups (p<0.05), and were 21% greater in smokers who received SDD compared with smokers who received placebo (p<0.05). Furthermore, month 9 PD reductions were 21-53% greater in non-smokers who received SDD compared with all other subgroups (p<0.01), and were 26% greater in smokers who received SDD compared with smokers who received placebo (p<0.05). CONCLUSION: Adjunctive SDD enhances therapeutic outcomes compared with SRP alone, resulting in clinical benefit in both smokers and non-smokers with CP. PMID- 15882220 TI - The effect of a dexibuprofen mouth rinse on experimental gingivitis in humans. AB - OBJECTIVES: The pharmacodynamic properties of ibuprofen are related nearly exclusively to the S(+)enantiomer (dexibuprofen). This study investigated the effect of a 1.5% dexibuprofen mouth rinse in an experimentally induced gingivitis. MATERIALS AND METHODS: The trial was a randomized, double-blinded, placebo-controlled, two-period and two-sequence parallel group cross-over study in 24 healthy volunteers aged 21-30 years (16 males, eight females). Customized guards were worn during tooth brushing to prevent any plaque removal from the experimental area (first and second pre-molars and molars in one upper quadrant). After 22 days of plaque accumulation, the mouth rinses (1.5% dexibuprofen and placebo) were administered under supervision three times daily (rinsing for 1 min. with 15 ml) for 8 days. The wash-out time between the two study periods was 14 days. Parameters evaluated at days 0, 7, 14, 22, and 30 were the Loe & Silness gingival index (GI) and the Quigley & Hein plaque index (QHI). Data were tested for treatment, period, and carry-over effects (parametric cross-over analysis). RESULTS: There was no statistically significant difference (p=0.240) in GI between placebo and dexibuprofen. However, the decrease in QHI was significantly greater (p=0.019) with dexibuprofen as compared with the placebo. CONCLUSION: In the present study, a 1.5% dexibuprofen mouth rinse had no effect on gingivitis whereas an anti-plaque effect was demonstrated. PMID- 15882221 TI - Relationship between maternal periodontal disease and low-birth-weight pre-term infants. AB - OBJECTIVES: The purpose of this study was to determine the influence of periodontal status on low-birth-weight pre-term delivery. MATERIAL AND METHODS: Ninety-six pregnant women were examined in their first, second and third trimester to record plaque scores, clinically assessed gingival inflammation and probing depth (mean depth and percentage of sites with depth of >3 mm). Binary logistic regression analyses were performed using SUDAAN 7.5 program. The type 1 (alpha) error established at 0.05 and an (alpha) error of 0.05-0.1 were considered nearly significant. RESULTS: The 96 women delivered 89 newborns: 16 were pre-term and seven of these were of low birth weight. There were seven miscarriages, all in the second trimester. No statistically significant association was found between gestational age and periodontal parameters. No significant relationship was found between low-weight delivery and plaque index measurements, although the association with gingival index was close to significant. A relationship was observed between low-weight birth and probing depth measurements, especially the percentage of sites of >3 mm depth, which was statistically significant (p=0.0038) even when gestational age was controlled for. CONCLUSIONS: According to these results, periodontal disease is a significant risk factor for low birth weight but not for pre-term delivery. PMID- 15882222 TI - Proportional distribution of the red complex and its individual pathogens after sample storage using the checkerboard DNA-DNA hybridization technique. AB - BACKGROUND: Information on the impact of sample storage prior to analysis by DNA methods is limited. AIMS: To investigate the effect of microbial sample storage on bacterial detection and proportional distribution of the red complex and its individual pathogens. MATERIAL AND METHODS: Subgingival plaque samples were analysed by (1) immediate processing, (2) after storage at +4 degrees C for 6 weeks, (3) after storage at -20 degrees C for 6 months or (4) after storage at 20 degrees C for 12 months using the checkerboard DNA-DNA hybridization. RESULTS: Proportional distribution of the red complex did not differ between the first three protocols. However, the total bacterial DNA for pathogens studied decreased significantly in protocols 3 and 4. Relative amounts of Tannerella forsythensis, Porphyromonas gingivalis and Treponema denticola remained stable in the second protocols and changed in an unpredictable way if stored for 6 or 12 months. CONCLUSIONS: Results from samples stored for maximum 6 months at -20 degrees C with high proportional amounts of the red complex and T. denticola may be used as an indicator of persistence. All bacterial samples for DNA extraction should be processed following a standardized storage protocol (i.e. samples stored at +4 degrees C for maximum 6 weeks) in order to get comparable qualitative and quantitative results for total DNA, bacterial complexes and individual pathogens. Most representative results are yielded if processing and hybridization could be performed immediately after sampling. PMID- 15882223 TI - Gingival tissue proteoglycan and chondroitin-4-sulphate levels in cyclosporin A induced gingival overgrowth and the effects of initial periodontal treatment. AB - OBJECTIVES: Cyclosporin A (CsA) is a potent immunosuppressive drug used in organ transplant patients to prevent graft rejection. CsA-induced gingival overgrowth is one of the side effects of this drug and its pathogenesis is still unclear. The present study was planned to comparatively analyse total proteoglycan (PG) and chondroitin-4-sulphate (C4S) levels in CsA-induced overgrown gingival tissue samples obtained before and after initial periodontal treatment and to compare these findings with the situation in healthy gingiva. MATERIAL AND METHODS: Gingival tissue samples were obtained from nine patients with CsA-induced gingival overgrowth before and 4 weeks after initial periodontal treatment including oral hygiene instruction and scaling and also from 10 healthy control subjects. Total PG and C4S levels were determined by biochemical techniques. PG levels were analysed using modified Bitter and Muir method. C4S assay was carried out using chondroitin sulphate lyase AC and chondroitin-6 sulphate sulphohydrolase enzymes. The results were tested statistically using non parametric tests. RESULTS: All clinical measurements in the CsA-induced gingival overgrowth group demonstrated significant reductions 4 weeks after initial periodontal treatment (p<0.05). There was no significant difference between the levels of baseline total PG in CsA-induced gingival overgrowth and healthy control groups (p>0.05). The gingival tissue levels of PG in CsA-induced gingival overgrowth group decreased significantly 4 weeks after treatment (p=0.043). Gingival tissue C4S levels in the overgrowth group were significantly higher than the healthy control group at baseline (p=0.000). C4S levels of the overgrowth group were significantly reduced after treatment (p=0.033), but these levels were still significantly higher than the healthy control group (p=0.000). CONCLUSION: The observed prominent increase in gingival tissue C4S levels may be interpreted as a sign of an increase in C4S synthesis in CsA-induced gingival overgrowth. Furthermore, remission of clinical inflammation by means of initial periodontal treatment had a positive effect on tissue levels of these extracellular matrix molecules. PMID- 15882224 TI - Lipoprotein-associated phospholipase A2 and plasma lipids in patients with destructive periodontal disease. AB - OBJECTIVES: Periodontitis is believed to be an independent risk factor of cardiovascular disease (CVD) and to be associated with a moderate systemic inflammatory reaction and hyperlipidaemia. Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) is an enzyme that has been shown to be a risk factor of CVD and that is involved in the degradation of the phospholipid mediator platelet activating factor (PAF), a potent mediator of inflammation. MATERIAL AND METHODS: In the present study, we measured concentrations of plasma lipids and plasma activity of Lp-PLA(2) in 32 patients (mean age 43+/-11 years) with moderate-to severe periodontitis before and 3 months after local treatment. RESULTS: Periodontal therapy resulted in a significant reduction of local inflammation and tissue destruction as reflected in reduced pocket depths and reduced bleeding indices. Pre- and post-treatment plasma lipid levels were (median and range, mmol/l): total cholesterol (C) 5.01 (3.94-7.15) and 4.91 (3.32-8.01); low-density lipoprotein-cholesterol (LDL-C) 3.14 (2.40-4.84) and 2.96 (1.39-5.04); HDL-C 1.27 (0.73-2.17) and 1.25 (0.74-2.55); triglycerides 1.37 (0.48-5.11) and 1.14 (0.38 792). Using the Wilcoxon's rank test, neither parameter showed a significant change. In contrast to the lacking response of plasma lipids, we observed a significant reduction in the activity of Lp-PLA(2). Local treatment lowered the enzyme activity by about 10% from 3.61+/-0.99 to 3.29+/-0.94 micromol/ml/h (mean+/-SD; p<0.001). The pre-treatment values of Lp-PLA(2) and LDL-C significantly correlated with clinical parameters of inflammation and periodontal destruction. CONCLUSION: This study indicates that treatment of periodontitis significantly reduces the serum activity of Lp-PLA(2), which is believed to be an independent cardiovascular risk factor. PMID- 15882225 TI - Ridge alterations following implant placement in fresh extraction sockets: an experimental study in the dog. AB - OBJECTIVE: To study dimensional alterations of the alveolar ridge that occurred following implant placement in fresh extraction sockets. MATERIAL AND METHODS: Five beagle dogs were included in the study. In both quadrants of the mandible, incisions were made in the crevice region of the third and fourth pre-molars. Buccal and minute lingual full-thickness flaps were elevated. The mesial root of the four pre-molars root was filled and the teeth were hemi-sected. Following flap elevation in (3)P(3) and (4)P(4) regions, the distal roots were removed. In the right jaw quadrants, implants with a sand blasted and acid etched (SLA) surface were placed in the fresh extraction sockets, while in the left jaws the corresponding sockets were left for spontaneous healing. The mesial roots were retained as surgical control teeth. After 3 months, the animals were examined clinically, sacrificed and tissue blocks containing the implant sites, the adjacent tooth sites (mesial root) and the edentulous socket sites were dissected, prepared for ground sectioning and examined in the microscope. RESULTS: At implant sites, the level of bone-to-implant contact (BC) was located 2.6+/-0.4 mm (buccal aspect) and 0.2+/-0.5 mm (lingual aspect) apical of the SLA level. At the edentulous sites, the mean vertical distance (V) between the marginal termination of the buccal and lingual bone walls was 2.2+/-0.9 mm. At the surgically treated tooth sites, the mean amount of attachment loss was 0.5+/ 0.5 mm (buccal) and 0.2+/-0.3 mm (lingual). CONCLUSIONS: Marked dimensional alterations had occurred in the edentulous ridge after 3 months of healing following the extraction of the distal root of mandibular pre-molars. The placement of an implant in the fresh extraction site obviously failed to prevent the re-modelling that occurred in the walls of the socket. The resulting height of the buccal and lingual walls at 3 months was similar at implants and edentulous sites and vertical bone loss was more pronounced at the buccal than at the lingual aspect of the ridge. It is suggested that the resorption of the socket walls that occurs following tooth removal must be considered in conjunction with implant placement in fresh extraction sockets. PMID- 15882226 TI - Improving patients' compliance with the treatment of periodontitis: a controlled study of behavioural intervention. AB - OBJECTIVES: This clinical trial study investigates whether a behavioural educational intervention based on the autoregulation theory can improve periodontitis patients' compliance with proper dental care at a 1-month follow up. MATERIAL AND METHODS: Thirty patients matched for gender (20 men), age (mean age=39 years) and education were randomly assigned to a control or an experimental treatment condition. In the control condition, patients received the regular treatment based on instruction of the proper prophylactic dental care. In the experimental treatment condition, patients received information about the symptoms of periodontitis, the causes, consequences and temporal course, and the types of effective strategy and they were requested to keep daily records of the effects of applying prophylactic dental care on their periodontitis symptoms. In both groups, plaque indices (PIs) (Silness & Loe 1964) were measured prior to treatment and at a 1-month follow-up. A self-report questionnaire also assessed the representation of periodontitis in all patients. RESULTS: A 2 (time of measurement: baseline versus follow-up) x 3 (PI localization) x 2 (experimental group) mixed-design anova computed on the PI reports a large effect of time, F(1, 28)=267.10, p<0.000, indicating that both groups improved from baseline (mean=1.73, SD=0.08) to the 1-month follow-up (mean=0.56, SD=0.06). It is important to point out that this analysis also revealed the expected Group x Time interaction, F(1, 28)=7.09, p<0.02, partial eta(2)=0.19, indicating that smaller PI were observed in the experimental group (mean=0.24, SD=0.14) than in the control group (mean=0.88, SD=0.38) at follow-up. Post hoc analyses showed that this pattern applies to the proximal and lingual PI but not to the vestibular PI. CONCLUSIONS: The present data show that the behavioural education intervention is (a) more effective than a classical intervention based on information and training about prophylactic techniques and (b) that it is effective in bringing most patients to normal levels of PI. For clinical practice, it suggests that better results can be obtained if (a) patients are taught a correct representation about periodontitis and (b) patients' sense of self-efficacy is developed through their own direct experience, by observing the effects of their behaviour on periodontitis symptoms. PMID- 15882227 TI - A method for the validation of a new calculus detection system. AB - BACKGROUND: Recently, pilot studies from our laboratory have shown that dental surfaces may be discriminated by the analysis of tip oscillations of an ultrasonic instrument, which possesses computerized calculus-detection features. For the evaluation of this smart detection system, its surface recognition qualities are of crucial importance. For in vivo studies, however, it proved to be difficult to verify the subgingival detection results. Therefore, it was necessary to develop a method, which allowed a reliable validation of surface recognition results of this new device. This evaluation method is described here. MATERIALS AND METHODS: Thirty extracted human teeth with subgingival calculus were embedded with plaster in a tray. To simulate subgingival pockets, dissected mucoperiostal porcine gingiva was sutured on the teeth. The thus-constructed dentition was mounted into a phantom head. A CCD-cam was attached with an intra oral X-ray mount to the teeth. The dentist scanned the pockets with the ultrasonic instrument, simultaneously videotaping the scanning path of the supragingival portion of the insert. At the same time, the signals of the modified ultrasound scaler were recorded. After the tooth was removed from the phantom head, the tip of the ultrasound scaler could be repositioned using the video sequences. The actual insert location on calculus or cementum was assessed and compared with the computer signals. The whole procedure was repeated a second time and the reproducibility of the evaluation method was estimated. RESULTS: A kappa value of 0.95 was attained for the evaluation method. CONCLUSION: The present experimental design allows the in vitro repositioning of an automated dental instrument for the detection of subgingival surfaces on the tooth following an in vitro phantom-head video recording of its intra-oral scanning movements. This method will be used for the verification of in vivo results of a new ultrasound-based surface detection system. PMID- 15882228 TI - Optimal rinsing time for intra-oral distribution (spread) of mouthwashes. AB - AIM: To investigate the intra-oral spread of an erythrosine mouthwash in relation to the rinsing period. MATERIAL AND METHODS: Thirty subjects were randomly divided into two equal groups and asked to rinse with 10 ml erythrosine mouthwash for cumulative periods of 15, 30 and 60 s (Group I) and 30, 60 and 90 s (Group II). Each rinsing session was followed by new plaque measurements. After rinsing plaque was finally assessed using the erythrosine mouthwash applied by means of a cotton swab. RESULTS: In Group I there was a difference observed between the 15 s rinsing period and those of 30 and 60 s. Explorative analysis for Group I suggested that differences in both jaws and approximal sites on both vestibular and lingual surfaces appeared to have contributed to the overall difference seen between 15 and 30 s rinsing periods. Also, pre-molars and front teeth seem to have contributed to this observed difference. No differences were noted between rinsing sessions and cotton swab application for Group II or Groups I+II combined. CONCLUSIONS: Rinsing for 30 s appeared to be sufficient for all plaque covered surfaces of the dentition to come into contact with the mouthwash. PMID- 15882229 TI - Clinical and microbiological evaluation of the effectiveness of the Nd:Yap laser for the initial treatment of adult periodontitis. A randomized controlled study. AB - BACKGROUND: Enhancement of the results obtained by scaling and planing is most often sought by using antimicrobial therapies. Laser beams have been shown to be bactericidal and could possibly target pathogens more effectively and with fewer compliance problems than antiseptic solutions. METHODS: Thirty subjects 20-60 years old presenting periodontal pockets at least 5 mm deep in each quadrant received initial periodontal treatment. The study had a split-mouth design. The control side (SRP) only received scaling and planing, and the test side (SRP+laser) was treated by both SRP and Nd:Yap (yttrium aluminum perovskite doped with neodym) laser. Clinical conditions were evaluated at day 0 and day 90 using the plaque index, gingival index, bleeding on probing, pocket probing depth, and clinical attachment level. Microbial sampling was also performed on days 0 and 90, and the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Treponema denticola was analysed by polymerase chain reaction in a commercial laboratory. Post-operative pain or discomfort was measured by the patient using a linear visual scale. Pearson's chi-squared test was used to compare bacterial presence. RESULTS: There was no statistically significant difference concerning clinical data between test and control groups at baseline. Both treatments enhanced the clinical situation compared to baseline; however, results were not significantly different between the two groups. T. forsythensis was the organism most numerous in both groups. Though initial treatment diminished the numbers of all the pathogens it did not do so statistically significantly. Differences between test and control groups were very small and bore no significance. Evaluation of the post-operative pain did not reveal any differences between the groups. CONCLUSIONS: Scaling and root planing was effective in reducing levels of plaque, inflammation, and bleeding upon probing. No additional advantage was achieved by using the Nd:Yap laser. PMID- 15882230 TI - Statistical methods for testing plaque removal efficacy in clinical trials. AB - OBJECTIVES: To evaluate the ability of different statistical approaches in finding a statistically significant difference in plaque removal efficiency between brushes in clinical trials. MATERIALS AND METHODS: The approaches, which are evaluated, concern the scores after brushing only, the difference in scores before and after brushing and the relative difference scores (i.e. score before minus score after brushing divided by the score before brushing). In each case the scores before brushing may be included as a covariate. Except for the relative difference scores, the power of the test statistics of the approaches has been compared by assuming a simple statistical model. These theoretical results have been compared with the numerical results of two particular clinical trials--one with a between-subject design and one with a within-subject design. RESULTS: The numerical results of these clinical trials show that the calculated p-values support the conclusions drawn from the statistical model, i.e. the power of the F-test is highest when evaluating the data after brushing with the data before brushing included as a covariate. Using the differences in scores before and after brushing--again with the data before brushing as a covariate--does not add additional power to the test. Omitting the data before brushing as a covariate only gives satisfactory results when the variance over the subjects or the error variance is zero, which in general is not the case. CONCLUSIONS: This investigation reveals that in general the approach of analysing the scores after brushing with the scores before brushing as a covariate yields the highest chance of finding a statistically significant difference between two brushes. PMID- 15882231 TI - Periodontal conditions and subgingival microflora in Down syndrome patients. A case-control study. AB - OBJECTIVES: The periodontal conditions and the subgingival microflora of children, adolescents and young adults (8-28 years old) with Down syndrome were investigated in the present cross-sectional study and compared with those of healthy individuals and subjects with cerebral palsy. MATERIAL AND METHODS: Seventy Down syndrome patients, 121 age-matched healthy individuals and 76 patients with cerebral palsy participated in the present study. Full-mouth recordings of clinical parameters (probing depth, probing attachment level, bleeding on probing, hygiene index) and the community periodontal index of treatment needs were assessed and subgingival plaque samples were taken from the Ramfjord teeth and analysed for 14 species using "checkerboard" DNA-DNA hybridization. RESULTS: Clinical indices of periodontal inflammation and treatment needs were statistically significant higher among Down syndrome patients compared with the other two groups (ANOVA, p=0.000). Important periodontal pathogens colonize these subjects earlier and at higher levels (chi squared test, p=0.000). DISCUSSION: Down syndrome patients display more severe periodontal destruction earlier, and heavier colonization with periodontal pathogens compared with age-matched healthy individuals and patients with cerebral palsy. PMID- 15882232 TI - Determinants of immunodiagnostic success in human ocular toxoplasmosis. AB - Ocular toxoplasmosis is a local manifestation of systemic infection in which Toxoplasma spreads into the eye, affecting mainly the posterior segment of the eye. Reactivation of the initial retinal condition presumably results from the rupture of quiescent parasitic cysts lying adjacent to pre-existing scars and may secondarily involve the choroid (leading to retinochoroiditis). Although the molecular mechanisms underlying host-parasite interaction are largely unknown, toxoplasmic retinochoroiditis usually remains a local event, and does not necessarily evoke a detectable systemic immune response. Local immunotolerance mechanisms may likewise confound attempts to confirm the clinical diagnosis by serology. Aqueous humour may be analysed for the presence of parasite DNA or of specific antibodies, but the DNA burden therein is low, and a more definite confirmation would require risky puncturing of the vitreous. Laboratory confirmation of the diagnosis is also frustrated by marked individual differences in the time elapsing between the onset of clinical symptoms and the activation of specific antibody production, resulting in a high proportion of false negative results. Whether a delay in the onset of local specific antibody production reflects immunotolerance in cases of congenital - but not obviously in those of acquired - infection remains an open question, but it could account for a relatively low confirmation rate in laboratory tests for local antibody production. Against this background, current diagnostic strategies need to be re evaluated with a view to future improvements. PMID- 15882233 TI - Isolates of Trichuris muris elicit different adaptive immune responses in their murine host. AB - The J and S isolates of Trichuris muris have different infection profiles in C57BL/6 mice; J worms are expelled, S worms survive to chronicity. Building on this, the ability of the J and S isolates to survive, and the quality of the immune response induced was explored in three different strains of mouse. The resistant BALB/c mouse mounted a strong Th2 response against both isolates, which were quickly expelled. The susceptible AKR host mounted a Th1 response and retained both isolates. Despite equivalent worm exposure, mesenteric lymph node cells from AKR mice infected with the S isolate produced significantly higher levels of IL-12 and the intestinal mastocytosis was reduced. IgG1 and IgG2a from S-infected AKR mice recognized low molecular weight antigens not recognized by J infected mice. Differential expulsion kinetics was observed in the slower responding C57BL/6 strain; J worms were expelled but S isolate worms were retained. Survival of the S isolate was again associated with elevated IL-12 and decreased Th2 responses. In resistant mouse strains, the outcome of infection is thus dominantly influenced by host genetics. However, in the slower-responding host, isolate-derived factors may play a role in shaping the quality of the adaptive immune response, thus influencing parasite survival. PMID- 15882234 TI - Host immune reactions and worm kinetics during the expulsion of Ascaris suum in pigs. AB - Pigs single inoculated with Ascaris suum eggs expel the majority of larvae between days 14 and 21 post inoculation (p.i.), but the role of the immune system in expulsion is unclear. To investigate the dynamics of immune responses before, during and after the expulsion of A. suum larvae, pigs inoculated with 10 000 A. suum eggs were sequentially necropsied. Ascaris suum gradually moved distally from days 10-14 p.i. and only a few larvae were left by day 21 p.i. Pronounced increases in mucosal A. suum-specific IgA antibody secreting cells (ASCs) were already found by day 10 p.i. especially in the proximal jejunum, while only small increases in parasite-specific IgM ASCs were observed by day 21 p.i. in both proximal and distal jejunum. No mucosal IgG ASC responses could be detected. Increases in systemic A. suum-specific IgG1, IgM and to a lesser extent IgA antibodies were observed, while IgG2 remained almost unchanged. The levels of eosinophils and mast cells in the small intestinal mucosa did not change throughout infection. The results demonstrate that both systemic and mucosal A. suum-specific effector mechanisms are strongly stimulated in A. suum single infections and indicate that mucosal IgA may be an important mediator in the expulsion of A. suum. PMID- 15882235 TI - Immuno-epidemiology of Ascaris lumbricoides infection in a high transmission community: antibody responses and their impact on current and future infection intensity. AB - The role of the humoral immune system in human infection with Ascaris lumbricoides remains unclear. This study documents an epidemiological investigation in a highly endemic community in Vietnam, whereby serum antibody levels were assessed before treatment and after a 6-month reinfection period. These data were examined by correlation with infection status using an age structured approach in an attempt to help shed light on the role of the humoral immune response. The first part of this study characterized levels of all serum antibody isotypes from the community in response to antigens of both adult and larval A. lumbricoides. Data were assessed in terms of their relation to host age and infection intensity with the aim to provide a broadly detailed account of immune responses to the parasite. In the second part, antibody responses to both life-stages of A. lumbricoides in serum samples collected before anthelmintic chemotherapy were analysed in relation to intensity of re-infection with the parasite 6 months following treatment. The results suggest that antibody responses may not confer protection from current infection or re-infection with A. lumbricoides and may not serve as reliable indicators of future infection intensity. Our results thereby lend support to the theory that immunity to A. lumbricoides may not be based on the humoral immune system. PMID- 15882236 TI - Schistosoma japonicum reveals distinct reactivity with antisera directed to proteases mediating host infection and invasion by cercariae of S. mansoni or S. haematobium. AB - Serine proteases released from the acetabular glands of cercariae, also known as cercarial elastases, are key enzymes in the penetration process of schistosomes through the skin of the final host. Antisera against these enzymes secreted from Schistosoma mansoni or S. haematobium reveal differences in the patterns of elastase expression among schistosome species and among different developmental stages of the larvae. Immunolocalization studies showed that antisera raised against the enzyme s28 protease react with S. mansoni, S. haematobium and also S. japonicum, in developing as well as mature cercariae and in both pre- and post acetabular glands. Antisera against the enzyme SmCE detect the respective antigen solely in the pre-acetabular glands. Remarkably, the SmCE-1a isoform is detectable with DNA-vaccinated mouse sera in S. mansoni and S. haematobium only, but is apparently absent from the acetabular glands of S. japonicum. These differences in immunoreactivity of cercarial enzymes may be related to the distinct infection process of S. japonicum. PMID- 15882237 TI - Personal view: victim blaming as management strategy for the gastroenterologist- a game theoretical approach. AB - BACKGROUND: A multitude of digestive diseases elude simple management strategies. Rather than admit failure of disease management, a gastroenterologist could resort to blaming patients for their own medical conditions. Blaming the patient constitutes an easy exit strategy for otherwise unsolvable disease conditions. AIM: To shed light on the problem of patient blaming in gastroenterology and provide means for its resolution. METHODS: The interaction between physician and patient can be formulated in terms of a non-zero-sum game between two adversaries. The outcomes associated with two behavioural strategies available to both adversaries are arranged in a two-by-two game matrix. RESULTS: Blaming the patient is characterized by the general game pattern of the 'prisoner's dilemma'. If the physician-patient interaction is restricted to one single event, patient blaming represents the management strategy of choice with the highest expected payoff under all foreseeable circumstances. If there is a high probability for repeated physician-patient interactions, however, a physician admitting and a patient accepting the limits of medical performance yield a dominant strategy. CONCLUSION: Only for single physician-patient encounters does a non-cooperative strategy of blaming one's adversary for a poor medical outcome yield the highest expected outcome. In the long run, the strategy of shifting blame becomes unproductive for both sides alike. PMID- 15882238 TI - A nationwide study of the incidence and 30-day mortality rate of pyogenic liver abscess in Denmark, 1977-2002. AB - BACKGROUND: Pyogenic liver abscess is a life-threatening disease. Accurate data on incidence and prognosis are important, but scarce. AIM: To examine changes in the incidence and 30-day mortality rate of patients with pyogenic liver abscess in Denmark. METHODS: Using nationwide administrative registers, we identified all patients diagnosed with pyogenic liver abscess in Denmark, 1977-2002, and their dates of death. We computed annual standardized incidence and 30-day mortality rates, and used Poisson regression to adjust gender-specific mortality rates for year-by-year differences in age at diagnosis. RESULTS: We identified 1448 patients with pyogenic liver abscess, of whom 54% were men. The crude incidence rate for the entire study period was 11.8 per 1,000,000 for men and 9.7 per 1,000,000 for women. Between 1977 and 2002, the incidence rate increased from 6 to 18 per 1,000,000 for men and from 8 to 12 per 1,000,000 for women. The cumulative 30-day mortality rate was 15% for men and 23% for women. The adjusted 30-day mortality rate decreased from 40% for men and 50% for women to around 10% for both genders. CONCLUSIONS: In this large nationwide study spanning a 26-year period, we found an increasing incidence rate and a decreasing mortality rate of pyogenic liver abscess. We believe that these changes are primarily explained by more sensitive diagnostic tools. PMID- 15882239 TI - Heartburn-dominant, uninvestigated dyspepsia: a comparison of 'PPI-start' and 'H2 RA-start' management strategies in primary care--the CADET-HR Study. AB - BACKGROUND: There are few data on empiric, stepped therapy for heartburn relief or subsequent relapse in primary care. AIMS: To compare heartburn relief produced by a proton pump inhibitor-start or an H(2)-receptor antagonist-start with step up therapy, as needed, followed by a treatment-free period to assess relapse. METHODS: Heartburn-dominant uninvestigated dyspepsia patients from 46 primary care centres were randomized to one of two active treatment strategies: omeprazole 20 mg daily (proton pump inhibitor-start) or ranitidine 150 mg bid (H2 receptor antagonist-start) for the first 4-8 weeks, stepping up to omeprazole 40 or 20 mg daily, respectively, for 4-8 weeks for persistent symptoms. Daily diaries documented heartburn relief (score < or = 3/7 on < or = of 7 prior days) and relapse (score > or = 4 on > or = 2 of 7 prior days). RESULTS: For 'proton pump inhibitor-start' (n = 196) vs. 'H2-receptor antagonist-start' (n = 194), respectively, heartburn relief occurred in 55.1% vs. 27.3% (P < 0.001) at 4 weeks and in 88.3% vs. 87.1% at 16 weeks. After therapy, 308 patients were heartburn free (159 vs. 149); median times to relapse were 8 vs. 9 days and cumulative relapse rates were 78.6% vs. 75.8%, respectively. CONCLUSIONS: An empiric 'proton pump inhibitor-start' strategy relieves heartburn more effectively than an 'H2 receptor antagonist-start' strategy up to 12 weeks but has no effect on subsequent relapse, which is rapid in most patients. PMID- 15882240 TI - Long-term use of acid suppression started inappropriately during hospitalization. AB - BACKGROUND: Practitioners routinely misuse acid suppression medications on general medical floors and inappropriately continue the drug at discharge. AIMS: To: (i) retrospectively study the appropriateness of acid suppression use on the general medical floors; (ii) characterize the patient population discharged on unnecessary acid suppression and (iii) evaluate whether patients discharged on unnecessary acid suppression continue the medicine long term. METHODS: Retrospective chart review of general medical patients admitted to an in-patient teaching service over 6 consecutive months. RESULTS: About 60% of patients lacked an indication for initiation of acid suppression and 34% of these patients were discharged on the medicine. The only independent predictor of continuation of acid suppression at discharge was longer length of stay. Multivariate analysis did not identify a characteristic distinguishing those patients discharged inappropriately on acid suppression. At 3 and 6 months of follow-up, 80% and 50% of patients, respectively, remained on acid suppression therapy without an appropriate indication. CONCLUSIONS: Our data verifies that practitioners routinely start general medical in-patients on acid suppression without an appropriate indication. Many of these prescriptions are continued at discharge for no apparent reason, leading to their long-term misuse. PMID- 15882241 TI - Polypharmacy and Crohn's disease. AB - BACKGROUND: Polypharmacy has not been defined for Crohn's disease. AIMS: To determine the prevalence of polypharmacy, factors associated with polypharmacy, and consequences of polypharmacy in a Crohn's disease population. METHODS: A review of 291 Crohn's disease patients was performed. Polypharmacy was defined as either minor (two to four medications) or major (> or = 5 medications). Clinical status was evaluated with the Harvey-Bradshaw index of disease activity (HBI) and the short inflammatory bowel disease questionnaire (SIBDQ). RESULTS: Major polypharmacy was identified in 50% of patients. Crohn's disease patients on less than two medications at the intake visit had an HBI of 3.6 compared with 5.4 and 6.0 in the minor and major polypharmacy groups (P < 0.05). Similarly, patients on less than two medications had an SIBDQ of 60.3 compared with 55.7 and 53.4 in the minor and major polypharmacy groups (P = 0.11). Predictors of polypharmacy included age > 40 years (OR 1.9), duration of disease > 10 years (OR 2.0), and female sex (OR 2.5). CONCLUSIONS: Polypharmacy is common in Crohn's disease and correlates with increased disease activity and decreased quality of life. Increasing age, increasing duration of disease, and female sex are associated with major polypharmacy. These findings emphasize the need for improved treatment algorithms to optimize Crohn's disease patient management. PMID- 15882242 TI - Experience of 5-aminosalicylate nephrotoxicity in the United Kingdom. AB - AIM: To study 5-aminosalicylate nephrotoxicity in patients with inflammatory bowel disease in the UK. METHODS: A detailed postal questionnaire was sent to all 1298 names in the British Society of Gastroenterology database and 290 consultant members of the Renal Association. The British Society of Gastroenterology reported new cases monthly, the Renal Association 6 monthly. Results were expressed as estimated glomerular filtration rate. RESULTS: Retrospective study: cases--British Society of Gastroenterology:Renal Association 202:87, aged 15-76 years. Median peak (range) creatinine (British Society of Gastroenterology:Renal Association) - 300:301 (78-1200) micromol/L. Prospective study - 59 cases, median age 52 years (M:F ratio: 47:12). Median pre-treatment estimated glomerular filtration rate: 76.9 (123.9-39), at diagnosis 28.4 (80.5-3.6, creatinine range: 92-1361 micromol/L), recovery 46.8 [111.2-end stage renal failure] mL/min/1.73 m2. Recovery of renal function was significantly improved for patients treated for < 12 months [n = 10, median recovery estimated glomerular filtration rate 70.5 (92-26.9) vs. > 12 months 38.4 (111.2-end stage renal failure) mL/min/1.73 m2, P = 0.028]. CONCLUSIONS: Regular monitoring of renal function may allow earlier detection of nephrotoxicity, particularly during the first year of therapy. Based on an inflammatory bowel disease prevalence in the United Kingdom of 412 x 10(5) with about 50% on treatment, we estimate that the incidence of clinical nephrotoxicity in patients taking 5-aminosalicylate therapy is approximately one in 4000 patients/year. PMID- 15882243 TI - A retrospective analysis of medical or surgical therapy in young patients with diverticulitis. AB - BACKGROUND: Acute diverticulitis is increasingly being recognized in younger patients, but its management remains controversial. AIM: To compare long-term outcomes of young patients treated with surgery vs. medical therapy for their first episode of diverticulitis. METHODS: A retrospective chart analysis at a university and an affiliated community hospital between 1991 and 2002 revealed 149 patients < or = 40 years of age with confirmed diverticulitis. Forty-nine patients (38 males, 11 females) were contacted at least 1 year after their first episode of diverticulitis. Outcomes were compared based on initial therapy- antibiotics or surgical resection. The groups were compared by outcomes, gender, age, white blood cell count, temperature and diet. RESULTS: Three (15%) of 20 surgical patients (mean follow-up 6.89 years), and 16 (55%) of 29 medical patients had a recurrence of diverticulitis (mean follow-up 5.72 years; P = 0.01). The treatment groups did not differ in age, white blood cell count, or temperature. CONCLUSIONS: (i) Surgical treatment is effective initial therapy but disease may recur in a minority of patients; (ii) medical treatment is less effective initial therapy, with recurrence in half of the patients; (iii) initial presentation is not a strong predictor of disease recurrence. PMID- 15882244 TI - Helicobacter pylori eradication does not cause reflux oesophagitis in functional dyspeptic patients: a randomized, investigator-blinded, placebo-controlled trial. AB - BACKGROUND: The protective role of Helicobacter pylori in gastro-oesophageal reflux disease has been widely discussed. AIM: To assess the risk of reflux oesophagitis in patients with functional dyspepsia after treatment for H. pylori infection. METHODS: A randomized, placebo-controlled, investigator-blinded trial was carried out on 157 functional dyspeptic patients. Patients were randomized to receive lansoprazole, amoxicillin and clarithromycin (antibiotic group) or lansoprazole and identical antibiotic placebos (control group). Upper gastrointestinal endoscopy was performed at baseline, 3 and 12 months after randomization. The primary aim was to detect the presence of reflux oesophagitis. Analyses were performed on an intention-to-treat basis. RESULTS: A total of 147 patients (94%) and 133 (85%) completed 3 months and 12 months follow-up, respectively. The eradication rate of H. pylori was 90% in the antibiotic group (74 of 82) and 1% (one of 75) in the control group. At 3 months, reflux oesophagitis was diagnosed in 3.7% (three of 82) in the antibiotic group and 4% (three of 75) in the control group (P > 0.2). At 12 months, diagnosis was established in five new cases within the first group and in four within the second (P > 0.2). No difference was found in heartburn symptoms. CONCLUSIONS: H. pylori eradication does not cause reflux oesophagitis in this western population of functional dyspeptic patients. PMID- 15882245 TI - Moxifloxacin-based strategies for first-line treatment of Helicobacter pylori infection. AB - BACKGROUND: Standard anti-Helicobacter pylori therapy may not achieve a satisfactory eradication rate. Fluoroquinolones, such as moxifloxacin, are safe and promising agents for H. pylori eradication. AIM: To compare the efficacy of two 1-week moxifloxacin-based H. pylori eradication regimens with two standard treatments. METHODS: Three hundred and twenty H. pylori-positive subjects were randomized into four groups to receive: moxifloxacin, amoxicillin, esomeprazole (Group MAE); moxifloxacin, tinidazole and esomeprazole (Group MTE); standard triple therapies with clarithromycin, amoxicillin and esomeprazole (Group CAE) or tinidazole (Group CTE) for 7 days. H. pylori status was re-assessed 6 weeks after the end of therapy by 13C urea breath test. RESULTS: Three hundred and twenty patients completed the efficacy analysis per protocol; H. pylori eradication rate in group MTE was 90% (72 of 80) and 92% (72 of 78), in group MAE was 88% (70 of 80) and 89%, (70 of 79) in Group CAE was 73% (58 of 80) and 78% (58 of 74), and in Group CTE was 75% (60 of 80) and 79% (60 of 76), respectively, in intention-to treat and in per protocol analyses. Eradication rates of moxifloxacin-based triple therapies were significantly higher than that observed using standard triple schemes. The incidence of side effects was significantly lower in moxifloxacin groups than in control groups. CONCLUSIONS: Seven-day moxifloxacin based triple therapies provide optimal eradication rates with a good compliance when compared with the standard triple therapy schemes. PMID- 15882246 TI - 7-day rescue therapy with ranitidine bismuth citrate after Helicobacter pylori treatment failure. AB - BACKGROUND: Quadruple rescue therapy requires a complex scheme with four drugs. AIM: To evaluate the efficacy of ranitidine bismuth citrate-tetracycline metronidazole rescue regimen, and to compare two different metronidazole dose schemes. METHODS: Prospective multicentre study including proton-pump inhibitor + clarithromycin + amoxicillin failures. Rescue regimen included two 7-day treatment: (i) ranitidine bismuth citrate (400 mg b.d.)-tetracycline (500 mg q.d.s.)-metronidazole (500 mg t.d.s.; RTM1); or (ii) the same regimen but with metronidazole 250 mg q.d.s. (RTM2). Eradication was confirmed with (13)C-urea breath test. RESULTS: A total of 150 patients were included (58 RTM1, 92 RTM2). All patients but two (one in each group) returned after treatment. About 86% in group RTM1 and 95% in RTM2 correctly took all the medications (P = 0.076). Per protocol eradication rates with RTM1 and RTM2 were 74 (95% CI: 60-84) and 69% (59 78). The intention-to-treat eradication rates were 64 (51-75) and 70% (59-78; P > 0.05). The type of regimen was not associated with eradication in the multivariate analysis. Adverse effects were more frequent with RTM1 (41%) than with RTM2 (30%; P > 0.05). CONCLUSION: Seven-day triple rescue therapy with ranitidine bismuth citrate-tetracycline-metronidazole is effective for Helicobacter pylori eradication, and represents an encouraging alternative to quadruple therapy, with the advantage of simplicity. The administration of metronidazole every 6 h (together with tetracycline), and at a low dose (250 mg), achieves similar efficacy and is probably associated with a better compliance and a lower incidence of adverse effects. PMID- 15882247 TI - Utility of the Health Belief Model in predicting compliance of screening in patients with chronic hepatitis B. AB - BACKGROUND: Regular surveillance is recommended for patients with chronic hepatitis B, to select candidates for anti-viral therapy and detect early complications. However, factors that determine compliance are not well studied. AIM: To determine the utility of the Health Belief Model in explaining non compliance, among a group of chronic hepatitis B patients for screening. METHODS: A total of 192 chronic hepatitis B patients who responded to advertisement for free screening took part in a telephonic interview study. Subjects were asked about the five constructs of the Health Belief Model, and factors associated with recent screening were analysed. RESULTS: The mean age of the subjects was 42.1 +/ 0.7 years; 77% white male, and 97% Chinese. About 108 patients (56%) had recent screening. At multivariate analysis, only the ability to remember date of follow up (OR: 4.37; 95% CI: 2.07-9.17) and the perception of having to wait a long time for venepuncture (OR: 0.38; 95% CI: 0.19-0.79) were significantly associated with recent screening. CONCLUSION: Future public health measures should include improving the logistics of follow-up procedures and providing reminders for screening to improve compliance. PMID- 15882248 TI - Probiotic supplementation improves tolerance to Helicobacter pylori eradication therapy--a placebo-controlled, double-blind randomized pilot study. AB - BACKGROUND: H. pylori is the major cause of chronic gastritis, and a risk factor for peptic ulcer and gastric cancer. AIM: To investigate the effect of probiotic supplementation on the tolerance and efficacy of H. pylori eradication treatment in a randomized, double-blind, placebo-controlled trial. METHODS: A total of 338 volunteers were screened for H. pylori infection. The eligibility criteria were met by 47 subjects whose H. pylori infection was verified at the outset and re evaluated after the treatment by the 13C-urea breath test and by enzyme immunoassay serology. The subjects were randomized to receive probiotic therapy (Lactobacillus rhamnosus GG, L. rhamnosusLC705, Bifidobacterium breve Bb99 and Propionibacterium freudenreichii ssp. shermanii JS) or a placebo during H. pylori eradication and for 3 weeks following the treatment, and recorded their daily symptoms in a standardized diary. RESULTS: When the frequencies of new or aggravated symptoms were evaluated, no significant differences were found between the two groups for individual symptoms. However, the probiotic group showed less treatment-related symptoms as measured by the total symptom score change (P = 0.038) throughout the H. pylori eradication therapy in contrast to the placebo group. The H. pylori eradication rate was non-significantly higher in the group receiving probiotic therapy (91% vs. 79%, P = 0.42). In this group the recovery of probiotic bacteria in the faeces increased significantly (P < 0.001). CONCLUSIONS: In this pilot study, probiotic supplementation did not diminish significantly the frequency of new or aggravated symptoms during H. pylori eradication. However, our data suggest an improved tolerance to the eradication treatment when total symptom severity was taken into account. Furthermore, the results show that probiotic bacteria are able to survive in the gastrointestinal tract despite the intensive antimicrobial therapy. PMID- 15882249 TI - Clinical features of type 1 autoimmune hepatitis in elderly Italian patients. AB - BACKGROUND: The usual onset of type 1 autoimmune hepatitis occurs at puberty or around menopause, whereas disease presentation in the advanced age is less often reported. AIM: To assess the clinical, immunological and histological features of Type 1 autoimmune hepatitis in elderly Italian patients. METHODS: We assessed, at diagnosis, the clinical and immunological features of 76 consecutive Italian patients with type 1 autoimmune hepatitis, focusing particularly on a subgroup of 20 patients presenting at > or = 65 years (females 95%, median age 72 years, range 65-82). RESULTS: In comparison with the younger group, at the time of autoimmune hepatitis diagnosis, elderly Italian patients are more often asymptomatic (25% vs. 7%; P = 0.04), are more frequently positive for antinuclear autoantibodies (95% vs. 52%; P = 0.0004) and HLA-DR4 (45% vs. 18%; P = 0.03); among the extra-hepatic manifestations, autoimmune thyroid disorders are prevalent in the elderly group (25% vs. 5%; P = 0.02). However, no difference was observed in the histological/biochemical expression of the liver disease and response to immunosuppression. CONCLUSIONS: In elderly Italian patients, autoimmune hepatitis has typical serological and genetic characteristics, is more frequently asymptomatic, although prognosis and response to therapy is similar to that of younger patients. As a concomitant autoimmune thyroid disorder is common, autoimmune hepatitis should be suspected and investigated in elderly patients with autoimmune thyroid disorder and abnormal liver function tests. PMID- 15882250 TI - Mycophenolate mofetil for the treatment of primary sclerosing cholangitis. PMID- 15882252 TI - Definition and classification of chronic kidney disease: a position statement from Kidney Disease: Improving Global Outcomes (KDIGO). AB - Chronic kidney disease (CKD) is a worldwide public health problem, with adverse outcomes of kidney failure, cardiovascular disease (CVD), and premature death. A simple definition and classification of kidney disease is necessary for international development and implementation of clinical practice guidelines. Kidney Disease: Improving Global Outcomes (KDIGO) conducted a survey and sponsored a controversies conference to (1) provide a clear understanding to both the nephrology and nonnephrology communities of the evidence base for the definition and classification recommended by Kidney Disease Quality Outcome Initiative (K/DOQI), (2) develop global consensus for the adoption of a simple definition and classification system, and (3) identify a collaborative research agenda and plan that would improve the evidence base and facilitate implementation of the definition and classification of CKD. The K/DOQI definition and classification were accepted, with clarifications. CKD is defined as kidney damage or glomerular filtration rate (GFR) <60 mL/min/1.73 m(2) for 3 months or more, irrespective of cause. Kidney damage in many kidney diseases can be ascertained by the presence of albuminuria, defined as albumin-to-creatinine ratio >30 mg/g in two of three spot urine specimens. GFR can be estimated from calibrated serum creatinine and estimating equations, such as the Modification of Diet in Renal Disease (MDRD) Study equation or the Cockcroft-Gault formula. Kidney disease severity is classified into five stages according to the level of GFR. Kidney disease treatment by dialysis and transplantation should be noted. Simple, uniform classifications of CKD by cause and by risks for kidney disease progression and CVD should be developed. PMID- 15882253 TI - Preeclampsia: a renal perspective. AB - Preeclampsia is a syndrome that affects 5% of all pregnancies, producing substantial maternal and perinatal morbidity and mortality. The aim of this review is to summarize our current understanding of the pathogenesis of preeclampsia with special emphasis on the recent discovery that circulating anti angiogenic proteins of placental origin may play an important role in the pathogenesis of proteinuria and hypertension of preeclampsia. PMID- 15882254 TI - Transcriptome analysis and kidney research: toward systems biology. AB - An enormous amount of data has been generated in kidney research using transcriptome analysis techniques. In this review article, we first describe briefly the principles and major characteristics of several of these techniques. We then summarize the progress in kidney research that has been made by using transcriptome analysis, emphasizing the experience gained and the lessons learned. Several technical issues regarding DNA microarray are highlighted because of the rapidly increased use of this technology. It appears clear from this brief survey that transcriptome analysis is an effective and important tool for question-driven exploratory science. To further enhance the power of this and other high throughput, as well as conventional approaches, in future studies of the kidney, we propose a multidimensional systems biology paradigm that integrates investigation at multiple levels of biologic regulation toward the goal of achieving a global understanding of physiology and pathophysiology. PMID- 15882255 TI - Role of coagulation factor Xa and protease-activated receptor 2 in human mesangial cell proliferation. AB - BACKGROUND: Fibrin deposition and mesangial cell proliferation are frequently observed in the active type of mesangioproliferative glomerulonephritis. Coagulation factors, such as factor V and factor Xa are colocalized with fibrin in the mesangial areas in active type of IgA nephropathy with mesangial cell proliferation. In this study, therefore, we studied the role of factor Xa and its receptor, protease-activated receptor 2 (PAR2) in mesangial cell proliferation and fibrin deposition, and examined ant-proliferative effects of a specific factor Xa inhibitor, DX-9065a, in cultured human mesangial cells. METHODS: To examine the effect of DX-9065a on the factor Xa-induced proliferation of cultured human mesangial cells, we measured thymidine incorporation and cell numbers. We also examined the effect of DX-9065a on extracellular regulated kinase (ERK) activation and fibrin production induced by factor Xa in human mesangial cells. RESULTS: Factor Xa increased [(3)H]-thymidine incorporation and cell numbers in a dose-dependent manner in mesangial cells, which was inhibited by DX-9065a. DX 9065a also suppressed factor Xa-triggered fibrin deposition on mesangial cell surface. Factor Xa induced the activation of ERK in mesangial cells and this activation was also completely inhibited by DX-9065a, but not inhibited by PAR1 antagonist. Factor Xa-induced cell proliferation and ERK activation were inhibited by PD98059. CONCLUSION: There results suggest that factor Xa can induce mesangial cell proliferation through the activation of ERK via PAR2 in mesangial cells and that PAR2 may play a crucial role in the cell proliferation induced by factor Xa. Our results implicate that DX-9065a may be a promising agent to regulate proliferation of mesangial cellss and inhibit the coagulation process in mesangium. PMID- 15882256 TI - ATR blockade reduces IFN-gamma production in lymphocytes in vivo and in vitro. AB - BACKGROUND: Type 1 angiotensin II (Ang II) receptor (AT(1)R) signaling induces proinflammatory responses. Recent studies suggest that T lymphocytes express AT(1)R; yet the effects of Ang II binding to AT(1)R on T cells are poorly understood. We examined the effect of AT(1)R blockade on release of the proinflammatory cytokine, interferon-gamma (IFN-gamma) by human lymphocytes in vivo and in vitro. METHODS: We used an AT(1)R blocker losartan in a randomized clinical trial in kidney transplant recipients over a 12-month period [AT(1)R blocker (N= 11) and control (N= 10)]. Peripheral blood lymphocytes, isolated from both cohorts, were analyzed by enzyme-linked immunosorbent spot assays (ELISPOT) analyses and real-time reverse transcription-polymerase chain reaction (RT-PCR) to enumerate IFN-gamma producing T cells and IFN-gamma mRNA levels. The effects of AT(1)R blockade in vitro were assessed using human alloreactive T cells and an IFN-gamma producing human cytotoxic T-lymphocyte line. Alloreactive T cells were treated with losartan or candesartan and enzyme-linked immunosorbant assay (ELISA) was used to measure IFN-gamma protein release. The cytotoxic T-lymphocyte line also was AT(1)R blocker-treated prior to determining IFN-gamma producing cells by intracellular cytokine staining. RESULTS: The AT(1)R blocker cohort had a significant decrease in IFN-gamma producing peripheral blood lymphocytes (P< or = 0.05 for each time point) and IFN-gamma mRNA levels (P= 0.01 vs. control patients). Losartan also decreased IFN-gamma production (P < 0.001) in purified alloreactive T cells in vitro as did candesartan. Moreover, Ang II amplified IFN gamma generation (P < 0.05) in alloreactive T cells while AT(1)R blocker treatment inhibited Ang II's effect (P < 0.04). AT(1)R blocker treatment furthermore also inhibited IFN-gamma production in the cytotoxic T-lymphocyte line. CONCLUSION: AT(1)R blockers may have a clinically relevant immunomodulatory role by blocking IFN-gamma production in T cells. PMID- 15882257 TI - Hepatocyte growth factor (HGF) modulates matrix turnover in human glomeruli. AB - BACKGROUND: The imbalance between synthesis and degradation of mesangial matrix causes glomerulosclerosis and leads to renal failure. Hepatocyte growth factor (HGF) has been shown to reduce the progression in murine models of chronic renal failure. The present study evaluated the effect of HGF on the extracellular matrix turnover and on c-met receptor in human glomeruli. METHODS: Human glomeruli microdissected from donor kidney biopsies before transplantation were incubated with culture media containing HGF (50 ng/mL). After 24 and 48 hours, the expression of c-met, (alpha2) IV collagen, transforming growth factor-beta (TGF-beta), metalloprotease (MMP) 2 and 9 and of the inhibitor of MMP-2, tissue inhibitors of metalloprotease-1 (TIMP-1), was evaluated by polymerase chain reaction (PCR). beta-actin was used as housekeeping gene. The production of collagen type IV and TGF-beta was evaluated by enzyme-linked immunosorbent assay (ELISA) and Western blotting and the activity of MMP by zymography. RESULTS: (alpha2) IV collagen, TGF-beta, and TIMP-1 mRNA levels were markedly decreased in glomeruli treated with HGF at 24 and 48 hours. The expression of c-met was up regulated by HGF treatment. HGF reduced the production of collagen type IV and TGF-beta. MMP-2 but not MMP-9 mRNA level was increased in HGF-treated glomeruli, although the gelatinolytic activity of the supernatant was not changed. By light microscopic examination kidney biopsies neither showed glomerular hypercellularity nor mesangial expansion. CONCLUSION: HGF reduced expression and synthesis of TGF-beta and collagen type IV and increased MMP-2 mRNA level in normal human glomeruli. These results suggest an antifibrotic effect of HGF on glomerular cells and may explain its beneficial role in glomerulosclerosis. PMID- 15882258 TI - Renal cyclooxygenase-2 in obese Zucker (fatty) rats. AB - BACKGROUND: Cyclooxygenase (COX) isoforms, COX-1 and COX-2, are involved in production of prostanoids in the kidney. Increases in renal COX-2 expression have been implicated in the pathophysiology of progressive renal injury, including type 1 diabetes. Thromboxane A(2) (TxA(2)) has been suggested as the key mediator of these effects resulting in up-regulation of prosclerotic cytokines and extracellular matrix proteins. Unlike type 1 diabetes, renal COX has not been studied in models of type 2 diabetes. METHODS: Renal cortical COX protein expression, and urinary excretion of stable metabolites of prostaglandin E(2) (PGE(2)) and TxA(2), in association with metabolic parameters, were determined in 4-and 12-week-old Zucker fatty rats (fa/fa rat) (ZDF4 and ZDF12), a model of type 2 diabetes, and in age-matched littermates with no metabolic defect (Zucker lean) (ZL4 and ZL12). RESULTS: Western blotting revealed increased COX-2 expression in ZDF4 as compared to ZL4 (245 +/- 130%) (P < 0.05). This increase in COX-2 was even more apparent in 12-week-old ZDF rats (650 +/- 120%) (P < 0.01). All groups of rats demonstrated COX-2-positive cells in typical cortical localizations [macula densa, thick ascending loop of Henle (TALH)]. In contrast to COX-2, COX-1 expression was 30% lower in ZDF12. These changes in COX expression were associated with enhanced urinary excretion of prostanoids, in parallel with the development of metabolic abnormalities. Moreover, increases in prostanoid excretion in ZDF12 were in part reduced by wortmannin (100 mug/kg), used as inhibitor of insulin signaling. CONCLUSION: Renal cortical COX-2 protein expression and function were increased in ZDF rats, as compared to controls, whereas COX-1 exhibited opposite regulation. The changes in COX-2 paralleled metabolic abnormalities, and were at least in part a four consequence of hyperinsulinemia. These abnormalities may play a role in renal pathophysiology in this model of type 2 diabetes. PMID- 15882259 TI - Dendrimer-enhanced MRI as a diagnostic and prognostic biomarker of sepsis-induced acute renal failure in aged mice. AB - BACKGROUND: Acute renal failure (ARF) induced by sepsis has a high mortality. In an aged mouse model of sepsis-induced ARF we have previously shown that renal injury occurs before serum creatinine is elevated. Development of a noninvasive biomarker that could diagnose renal dysfunction early in sepsis and monitor the response to therapy would be very valuable. METHODS: We performed magnetic resonance imaging (MRI) with gadolinium-based G4 dendrimer intravenous contrast in a fluid- and antibiotic-treated cecal ligation and puncture (CLP) sepsis model in aged mice. Imaging was also performed in a mouse volume depletion model and in models of ARF induced by ischemia/reperfusion (I/R) and cisplatin. RESULTS: Twenty hours post-CLP, aged mice had a distinct pattern of renal injury using dendrimer-enhanced MRI. This pattern was different from renal injury induced by either cisplatin or I/R. Prerenal azotemia induced by volume depletion was distinguished from sepsis by dendrimer-enhanced MRI. Dendrimer-enhanced MRI detected renal dysfunction 6 hours post-CLP, a time when serum creatinine was still normal. Ethyl pyruvate reversed the renal dysfunction detected by dendrimer enhanced MRI at 20 hours, but not at 6 hours post-CLP. The appearance of renal dysfunction on dendrimer-enhanced MRI at 6 hours post-CLP predicted the length of survival. CONCLUSION: Dendrimer-enhanced MRI is a novel biomarker that provides information for the early diagnosis, drug responsiveness, and prognosis of sepsis induced ARF. PMID- 15882260 TI - T-cell transcriptome analysis points up a thymic disorder in idiopathic nephrotic syndrome. AB - BACKGROUND: Idiopathic nephrotic syndrome is a proteinuric disease secondary to the release of a nonidentified circulating glomerular permeability factor by T cells. Because specificities of T-cell activation in idiopathic nephrotic syndrome remain unknown, we evaluated transcriptional activation of T cells in nephrotic patients during proteinuria. METHODS: Transcriptomes of CD2+ cells were analyzed by serial analysis of gene expression (SAGE) in a nephrotic child during proteinuria relapse and after remission, away from any immunosuppressive treatment. Expression of specific transcripts overexpressed during proteinuria relapse was compared by reverse transcription-polymerase chain reaction (RT-PCR) in CD2+ cells from 11 nephrotic patients during relapse and remission and 11 non nephrotic patients during infection and after recovery. RESULTS: Differential analysis of CD2+ cell transcriptome identified >200 mRNA tags overexpressed during proteinuria relapse, including many T-cell markers. RT-PCR analysis of expression of specific transcripts indicated that (1) under remission conditions, nephrotic children displayed induction of four transcripts, including IKBKB, and repression of NFKBIA as compared to non nephrotic children after recovery, and (2) proteinuria relapse was associated with induction of L-selectin and T lymphocyte maturation-associated protein, two markers of T-cell differentiation and recent emigrant/naive T cells. CONCLUSION: Results indicate that circulating T cells from relapsing nephrotic patients include a significant population of low mature cells while those from nephrotic patients in remission are characterized by constitutive activation of nuclear factor-kappaB (NF-kappaB), altogether suggesting a thymic dysregulation of apoptosis in nephrotic patients. PMID- 15882261 TI - DNA vaccination with naked DNA encoding MCP-1 and RANTES protects against renal injury in adriamycin nephropathy. AB - BACKGROUND: We have previously shown that monocyte chemoattractant protein-1 (MCP 1) and regulated upon activation, normal T-cell expressed and secreted (RANTES) are significantly increased in renal cortex in adriamycin nephropathy. In this study, we tested the effect of DNA vaccination encoding the C-C chemokines MCP-1 and RANTES in a rat model of adriamycin nephropathy. METHODS: Both reverse transcription-polymerase chain reaction (RT-PCR) products of MCP-1 and RANTES used as constructs were cloned into a pTarget vector for naked DNA vaccination. Two hundred micrograms of DNA was injected into the tibialis anterior muscle four times at weekly intervals. One week after the last DNA vaccination, rats received adriamycin. All animals were sacrificed 4 weeks after adriamycin administration. Changes in renal function and histologic features were assessed. Enzyme-linked immunosorbent assay (ELISA) and Western blot were used for autoantibody determination. Antibody specificity was assessed in in vitro transmigration assays. RESULTS: Chemokine DNA vaccination significantly reduced proteinuria (P < 0.05) and ameliorated creatinine clearance (P < 0.05) at 2, 3, and 4 weeks after adriamycin administration. Morphometric analysis showed less glomerular sclerosis (P < 0.001) and interstitial infiltrates (P < 0.005) in chemokine DNA vaccination group compared with control groups. Anti-MCP-1 and RANTES autoantibodies were detected in higher concentrations in chemokine DNA vaccinated rats than in control rats (P < 0.001) and serum from vaccinated rats blocked T-cell transmigration to MCP-1 and RANTES. CONCLUSION: In this study, we have shown that naked DNA vaccination against MCP-1 and RANTES ameliorates the progression of renal disease in the rat adriamycin nephropathy model of chronic proteinuric renal disease. The protective mechanism may involve the production of autoantibodies against MCP-1 and RANTES. PMID- 15882262 TI - Small bowel cyclooxygenase 2 (COX-2) expression in patients with IgA nephropathy. AB - BACKGROUND: Clinical manifestation of IgA nephropathy (IgAN) strikingly occurs after respiratory tract infections. An intestinal inflammation has also been described. We hypothesized that the intestinal inflammation should manifest itself as an increase in inflammatory cells and mucosal cyclooxygenase 2 (COX-2) expression. METHODS: By using immunohistochemistry, we determined the phenotype and quantity of inflammatory cells in duodenal biopsy specimens from 17 IgAN patients. Control material comprised 18 patients undergoing gastroscopy because of dyspepsia. RESULTS: All the biopsy specimens disclosed normal villous architecture. In IgAN, CD3(+) cells and COX-2-positive cells were significantly increased and J chain-producing plasma cells were significantly decreased. CD3(+) cells coexpressed COX-2 protein and COX-2-positive cells also expressed CD45RO antigen. The number of lymphocytes correlated significantly with serum IgA and COX-2-expression with serum IgA and the degree of hematuria. COX-2-positive subepithelial fibroblasts were a conspicuous finding in IgAN. In CD68(+) and CD15(+) cells, a significant increase was seen. Many of these cells also expressed COX-2 protein. CD15(+) positivity correlated significantly with proteinuria in IgAN. CONCLUSION: Our results indicate that small bowel inflammation in IgAN shows itself as an increased number of mucosal inflammatory cells. However, polymeric IgA production is significantly decreased. An increased mucosal COX-2 expression suggests activation of the inflammatory cells and the degree of inflammation significantly correlates with serum IgA and the amount of proteinuria and hematuria. Subepithelial fibroblasts seem also to be involved in the inflammatory reaction. PMID- 15882263 TI - Triglyceride accumulation in injured renal tubular cells: alterations in both synthetic and catabolic pathways. AB - BACKGROUND: Triglycerides can accumulate in injured tissues, a process thought to represent flux of excess, cytotoxic, free fatty acids into nontoxic triglyceride storage pools. However, this view may be overly simplistic, given that multiple pathways may impact triglyceride levels. This study sought new insights into this issue. METHODS: Cultured human proximal tubule [human kidney-2 (HK-2)] cells or in vivo kidney were subjected to injuries known to increase triglyceride levels approximately three- to fourfold [HK-2 cells antimycin A-induced mitochondrial blockade; in vivo glycerol-induced rhabdomyolysis; endotoxemia). Six reverse transcription-polymerase chain reactions (RT-PCRs) were used to monitor mouse/human mRNAs for renal fatty acid transport protein (FATP2), or triglyceride synthesizing enzymes (acyl-coenzyme A:diacylglycerol acyltransferases DGAT1 and DGAT2). Fatty acid synthase (FAS) and FATP2 were gauged by Western blot. FAS, FATP2, mitochondrial respiration, and phospholipase A2 (PLA(2)) effects on cell triglyceride accumulation were probed. Finally, tissue lipase activity was assessed. RESULTS: Antimycin A up-regulated multiple determinants of HK-2 cell triglyceride formation, including FATP2, FAS, DGAT1, and DGAT2 (proteins and/or mRNAs). However, neither FAS- nor FATP2-inhibition eliminated antimycin A-induced triglyceride loading, indicating the latter's multifactorial basis. PLA(2) activity increased FFA and triglyceride content. Rhabdomyolysis and endotoxemia altered multiple triglyceride homeostatic mechanisms. However, these changes were model-dependent and did not closely parallel those in HK-2 cells. Lipase activity signficantly fell (glycerol) or rose (endotoxemia) with different forms of tissue damage. CONCLUSION: Injury-induced triglyceride accumulation stems from multiple, and disease-specific, changes in triglyceride synthetic and degradative pathways. Simple flux of excess FFAs into triglyceride pools is an overly simplistic view of the post-injury-triglyceride loading state. PMID- 15882264 TI - Geranylgeranylacetone ameliorates ischemic acute renal failure via induction of Hsp70. AB - BACKGROUND: Heat shock proteins (HSPs) are well known as cytoprotective proteins. Geranylgeranylacetone (GGA), an antiulcer agent, has recently been shown to induce Hsp70. This study was performed to investigate the renoprotective properties of GGA. METHODS: The effect of GGA on the induction of the major HSPs (Hsp90, Hsp70, Hsc70, Hsp60, and Hsp32) was studied in the rat kidney or rat primary cultures of tubular epithelial cells (R-TECs) by Western blot. Localization of Hsp70 was determined by immunohistochemistry. The renoprotective effects of GGA were studied using a rat model of ischemia/reperfusion (I/R) injury. GGA (400 mg/kg), GGA with quercetin pretreatment (100 mg/kg), or a vehicle was given to rats 24 hours and again 1 hour prior to the induction of I/R injury. Rats were sacrificed at 24 hours after reperfusion. Histologic analyses and terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) assay were performed. Blood urea nitrogen (BUN) and serum creatinine was also measured. The cytoprotective properties of GGA were also studied in vitro by treating R-TECs with GGA (10 mumol/L) or a vehicle, followed by incubation in culture medium with oxidative stress condition (0.5 mmol/L hydrogen peroxide) or ischemic condition (2 nmol/L NaCN and 20 mmol/L 2-deoxyglucose in the absence of medium glucose). RESULTS: Oral administration of GGA induced Hsp70 expression in the kidney (which peaked at 24 hours) but did not induce Hsp90, Hsc70, Hsp60, or Hsp32. The induction of Hsp70 was blocked by quercetin. Immunohistochemistry showed that Hsp70 was localized mainly in the tubular epithelial cells. Preconditioning rats with GGA significantly decreased BUN and serum creatinine levels after I/R injury. Histologic examination revealed that GGA significantly attenuated tubular damage and macrophage infiltration. The number of TUNEL-positive cells also decreased significantly in the GGA group. Quercetin, an inhibitor of Hsp70 induction, eliminated these renoprotective effects of GGA. In in vitro study, GGA-induced Hsp70 in R-TECs, which peaked at 2 to 4 hours. Both oxidative stress and ischemic stimuli induced apoptosis in R-TECs. GGA significantly suppressed the number of apoptotic cells in both conditions. CONCLUSION: The results support the hypothesis that GGA induces Hsp70, protects tubular epithelial cells from apoptosis, and thus ameliorates tubular damage by I/R injury. The present study suggests that GGA would be a useful tool in treating acute renal failure or preventing transplanted kidney damage in the clinical setting. PMID- 15882265 TI - Multifunctionality of PAI-1 in fibrogenesis: evidence from obstructive nephropathy in PAI-1-overexpressing mice. AB - BACKGROUND: Plasminogen activator inhibitor-1 (PAI-1) has been implicated in the pathogenesis of chronic kidney disease based on its up-regulated expression and on the beneficial effects of PAI-1 inhibition or depletion in experimental models. PAI-1 is a multifunctional protein and the mechanisms that account for its profibrotic effects have not been fully elucidated. METHODS: The present study was designed to investigate PAI-1-dependent fibrogenic pathways by comparing the unilateral ureteral obstruction model (UUO) (days 3, 7, and 14) in PAI-1-overexpressing mice (PAI-1 tg) to wild-type mice, both on a C57BL6 background. RESULTS: Following UUO, total kidney PAI-1 mRNA and/or protein levels were significantly higher in the PAI-1 tg mice (N= 6 to 8/group) and fibrosis severity was significantly worse (days 3, 7, and 14), measured both as Sirius red positive interstitial area (e.g., 10 +/- 3.2% vs. 4.5 +/- 1.0%) (day 14) and total kidney collagen (e.g., 11.1 +/- 1.7 vs. 6.2 +/- 1.3 microg/mg) (day 14). By day 14, the expression of two normal tubular proteins, E-cadherin and Ksp cadherin, were significantly lower in the PAI-1 tg mice (3.2 +/- 0.5% vs. 11.7 +/ 5.9% and 2.6 +/- 1.6) vs. 6.2 +/- 0.8%, respectively), implying more extensive tubular damage. At least four fibrogenic pathways were differentially expressed in the PAI-1 tg mice. First, interstitial macrophage recruitment was more intense (P < 0.05 days 3 and 14). Second, interstitial myofibroblast density was greater (P < 0.05 days 3 and 7) despite similar numbers of proliferating tubulointerstitial cells. Third, transforming growth factor-beta1 (TGF-beta1) and collagen I mRNA were significantly higher. Finally, urokinase activity was significantly lower (P < 0.05 days 7 and 14) despite similar mRNA levels. Gene microarray studies documented that that the deletion of this single profibrotic gene had far-reaching consequences on renal cellular responses to chronic injury. CONCLUSION: These data provide further evidence that PAI-1 is directly involved in interstitial fibrosis and tubular damage via two primary overlapping mechanisms: early effects on interstitial cell recruitment and late effects associated with decreased urokinase activity. PMID- 15882266 TI - Nephrin and podocin dissociate at the onset of proteinuria in experimental membranous nephropathy. AB - BACKGROUND: The slit diaphragm plays a critical role in maintaining the barrier function of the glomerular capillary wall. The pathogenic mechanism of proteinuria in membranous nephropathy remains uncertain. This study was undertaken to analyze the pathogenic role of slit diaphragm in proteinuria in experimental membranous nephropathy. METHODS: The expression and the localization of slit diaphragm-associated molecules (nephrin, podocin, and CD2AP) and other podocyte-associated molecules (podocalyxin and alpha(3) integrin) in passive and active Heymann nephritis were analyzed by immunofluorescence and Western blot analysis. The interaction of slit diaphragm-associated molecules was investigated by the dual-labeling immunofluorescence method. The mRNA expression of these molecules was also analyzed. RESULTS: Shifts in nephrin and podocin staining patterns, from linear to granular, were detected in the early stages of passive Heymann nephritis. These shifts were not parallel, and the dissociation of these molecules was detected by the dual-labeling immunofluorescence method in passive and active Heymann nephritis. Western blot analyses with sequentially solubilized materials indicated that the nephrin-rich fraction changed from being partly detergent-resistant to being predominantly detergent-soluble. This change did not occur with podocin. Nephrin excreted into urine was already detected in the early stages of passive Heymann nephritis. Decreased mRNA expression of nephrin and podocin was observed before the onset of proteinuria. By contrast, no extensive change in the expression of alpha(3) integrin was observed in this study. CONCLUSION: Nephrin is dissociated from podocin and excreted into urine in the early stages of Heymann nephritis. The reduced expression of nephrin and podocin, along with their dissociation, may contribute to the development of proteinuria in Heymann nephritis. PMID- 15882267 TI - Up-regulation of tissue factor activity on human proximal tubular epithelial cells in response to Shiga toxin. AB - BACKGROUND: The pathophysiology of hemolytic uremic syndrome (HUS) is incompletely established. Based on clinical studies demonstrating the presence of prothrombotic plasma markers in patients with HUS, we hypothesized that Shiga toxin might cause activation of the coagulation pathway by augmenting tissue factor, the major initiator of coagulation. METHODS: Human proximal tubular epithelial cells (PTECs) [human kidney-2 (HK-2 cells)] were exposed to Shiga toxin-1, and expression of tissue factor, cell detachment, protein synthesis, caspase-3 activity, and Shiga toxin-1 binding were examined. Results. HK-2 cells expressed constitutive surface tissue factor activity and increased their tissue factor expression upon exposure to Shiga toxin-1. Shiga toxin-1 bound to HK-2 cells and inhibited protein synthesis. The up-regulation of tissue factor was dose- and time-dependent and strongly correlated with cell detachment and increase in caspase-3 activity caused by Shiga toxin-1 exposure. A general caspase inhibitor simultaneously inhibited HK-2 cell detachment and tissue factor up-regulation while mutant Shiga toxin-1 neither caused cell detachment, protein synthesis inhibition, nor increase in tissue factor activity. Tissue factor activity elicited by Shiga toxin-1 was abrogated by a monoclonal antitissue factor antibody. Calphostin C, a protein kinase C (PKC) inhibitor, partially blocked tissue factor up-regulation, indicating possible involvement of PKC dependent mechanism. CONCLUSION: These data, taken together, suggest a strong link between Shiga toxin-induced up-regulation of tissue factor activity, cytotoxicity, and apoptosis in HK-2 cells. The proximal tubule is a target of Shiga toxin in HUS, and it seems plausible that injured proximal tubular cells trigger the activation of the coagulation system, the formation of intrarenal platelet-fibrin thrombi, and the development of acute renal failure in HUS. PMID- 15882268 TI - Progression of secondary hyperparathyroidism involves deregulation of genes related to DNA and RNA stability. AB - BACKGROUND: Renal secondary hyperparathyroidism in its late stages becomes autonomous, so excessive parathyroid hormone (PTH) secretion no longer responds to physiologic stimuli or to aggressive medical treatment. METHODS: To gain molecular understanding of progression of renal secondary hyperparathyroidism, normal and hyperplastic parathyroid tissue with diffuse and nodular growth were analyzed. The results were also compared to parathyroid adenomas. The analysis was performed by high-density oligonucleotide microarray and bidirectional subtraction library. RESULTS: Analysis of the DNA arrays found 16 overexpressed and 132 repressed genes in the nodules while the subtraction library produced 34 overexpressed and 40 repressed genes. The differentially expressed genes between diffuse and nodular samples included some related to DNA stability and repair (TALDO1, PRDX2, DDB1, XRCC1, and POLB), RNA stability and degradation (OASL and AUF1), protein synthesis and processing (PFDN5, HSPD1, and NACA), cell growth (CDC25C and GRPR), and tumorigenesis and cell cycle (VIL2 and TPD52). CONCLUSION: According to the function described for the deregulated genes, when secondary hyperparathyroidism becomes autonomous and refractory to treatment, RNA degradation may be increased while DNA integrity may be compromised. These two mechanisms, combined with deregulation of genes related to growth and differentiation show the complex pathway of parathyroid glands' evolution in renal hyperparathyroidism and may explain the large amount of molecular cytogenetic aberrations found in refractory hyperparathyroidism. Considering that some of the genes with altered expression in nodular hyperplasia lead to irreversible consequences in the genomic integrity of the cells, an adequate and early management of the secondary hyperparathyroidism of chronic kidney disease becomes mandatory. PMID- 15882269 TI - Impaired EDHF-mediated vasodilation and function of endothelial Ca-activated K channels in uremic rats. AB - BACKGROUND: Chronic renal failure (CRF) is associated with increased cardiovascular morbidity, abnormal arterial tone, and endothelial dysfunction. Ca(2+)-activated K(+)-channels (K(Ca)) are important regulators of endothelial function by controlling endothelial hyperpolarization and thus endothelium derived hyperpolarizing factor (EDHF)-mediated vasodilations. Here we tested the hypothesis whether an altered function of endothelial K(Ca) and diminished EDHF mediated vasodilation contribute to the endothelial dysfunction in the rat remnant kidney model of chronic renal failure. METHODS: Functional expression of endothelial K(Ca) currents and endothelium-dependent vasodilations in rat carotid arteries were assessed by using patch-clamp techniques, single-cell reverse transcription-polymerase chain reaction (RT-PCR), and a pressure myograph 8 weeks after either subtotal 5/6 nephrectomy in normotensive or hypertensive, or sham operated rats. RESULTS: Acetylcholine (ACh)-induced EDHF-mediated vasodilations were present in sham-operated rats, but almost absent in both normotensive 5/6 nephrectomy rats and hypertensive 5/6 nephrectomy rats. In experiments without blocking nitric oxide/prostacyclin synthesis, endothelium-dependent vasodilation to ACh was significantly reduced in both normotensive 5/6 nephrectomy rats and hypertensive 5/6 nephrectomy rats. In sham-operated rats, 1-ethyl-2 benzimidazolinone (1-EBIO), a selective opener of endothelial small and intermediate K(Ca), induced a substantial EDHF-mediated vasodilation, which was greatly reduced in hypertensive 5/6 nephrectomy rats and in normotensive 5/6 nephrectomy rats. In patch-clamp experiments, mean K(Ca) currents were significantly reduced in endothelial cells from hypertensive 5/6 nephrectomy rats and normotensive 5/6 nephrectomy rats when compared to sham-operated rats. Concordantly, single-cell reverse-transcription-polymerase chain reaction (RT PCR) analysis revealed a greatly reduced frequency of endothelial cells expressing the K(Ca) genes, SKCa3 and IKCa1 in 5/6 nephrectomy rats compared to sham-operated rats. CONCLUSION: Experimental CRF leads to a loss of EDHF-type vasodilation which was caused at least in part by an impaired functional expression of endothelial hyperpolarizing K(Ca). The loss of EDHF-type vasodilation may contribute to endothelial dysfunction and abnormal arterial tone in CRF. PMID- 15882270 TI - The antioxidant N-acetylcysteine prevents accelerated atherosclerosis in uremic apolipoprotein E knockout mice. AB - BACKGROUND: Cardiovascular disease is the most frequent cause of mortality in chronic renal failure (CRF). Therefore, it is important to identify appropriate treatment measures. The antioxidant N-acetylcysteine (NAC) has been shown to reduce cardiovascular events in hemodialysis patients. Here we examine a possible direct effect of NAC supplementation on uremia-enhanced atherosclerosis in apolipoprotein E-deficient (apoE(-/-)) mice. METHODS: Uremia was induced surgically in 8-week-old female apoE(-/-) mice. Two weeks after creation of CRF mice were randomized to receive either NAC (daily oral gavage with 200 mg/kg for 8 weeks) or placebo. They were compared to a control group of sham-operated apoE( /-) mice receiving placebo. After 8 weeks of treatment, the mice were sacrificed, and the cross-section surface area of atherosclerotic plaques was measured in aortic root and descending aorta. RESULTS: At 10 weeks following surgery, atherosclerotic lesions were significantly larger in uremic apoE(-/-) mice than in nonuremic controls. This accelerated atherosclerosis was associated with an increase in aortic nitrotyrosine expression and collagen plaque content. NAC treatment inhibited the progression of atherosclerotic lesions and plaque collagen content compared with placebo treatment. In addition, plaques from NAC treated uremic animals showed a significant decrease in nitrotyrosine expression whereas the degree of macrophage infiltration was comparable in both uremic groups. There was no difference in mean arterial blood pressure between the three groups. CONCLUSION: We show for the first time that the antioxidant NAC is capable of reducing atheroma progression, in an animal model of uremia-enhanced atherosclerosis, probably via a decrease in oxidative stress. PMID- 15882271 TI - Role of calcification inhibitors in the pathogenesis of vascular calcification in chronic kidney disease (CKD). AB - BACKGROUND: The majority of patients with chronic kidney disease (CKD) have excessive vascular calcification; however, most studies demonstrate that a subset of CKD patients do not have, nor develop, vascular calcification despite similar exposure to the uremic environment. This suggests protective mechanisms, or naturally occurring inhibitors, of calcification may be important. METHODS: In order to determine the role of three inhibitors, fetuin-A, matrix gla protein (MGP), and osteoprotegerin (OPG) in the vascular calcification observed in patients with CKD-5, we (1) measured serum levels of these inhibitors and compared the levels to calcification assessed by computed tomography (CT); (2) examined arteries from CKD-5 patients by immunostaining for these inhibitors; and (3) examined the expression and effect of these inhibitors in cultured bovine vascular smooth muscle cells (BVSMCs) incubated in serum pooled from uremic patients compared to healthy controls. RESULTS: There was a negative correlation of coronary artery calcification scores with serum fetuin-A levels (r=-0.30, P= 0.034) and a positive association with OPG levels (r= 0.29, P= 0.045). There was increasing immunostaining for both fetuin-A and MGP in arteries with increasing calcification graded semiquantitatively (P < 0.003). In vitro, fetuin-A added to mineralizing BVSMCs inhibited mineralization (P < 0.001). Compared to normal serum, BVSMCs incubated with uremic serum had a progressive increase in MGP expression with mineralization (P < 0.001) and increased expression of OPG in BVSMCs (P < 0.04). CONCLUSION: These data demonstrate that fetuin-A, OPG, and MGP play an important role in the pathogenesis of uremic vascular calcification. PMID- 15882272 TI - Validation of quantitative BOLD MRI measurements in kidney: application to unilateral ureteral obstruction. AB - BACKGROUND: Blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) provides a measure of deoxyhemoglobin content and therefore an indirect measure of the partial oxygen pressure (pO(2)). The main purpose of this study was to examine the relationship between the apparent relaxation rate (R2*) in the pig kidney by BOLD imaging and renal tissue pO(2) levels measured directly by oxygen-sensitive microelectrodes. Second, BOLD imaging was applied to kidneys in pigs subjected to acute unilateral ureteral obstruction (UUO) to examine whether this condition is associated with changes in intrarenal oxygenation. METHODS: Oxygen-sensitive microelectrodes were inserted in the cortex and medulla of pig kidneys (N= 6). Different arterial and intrarenal levels of pO(2) were obtained by stepwise changing the oxygen-to-nitrogen ratio supplied by a respirator. Simultaneous BOLD MRI measurements using an R2*-sensitive Echo Planar Imaging (EPI) sequence were performed on the contralateral kidney. In another group of pigs (N= 3) BOLD imaging was performed following 24 hours of UUO. RESULTS: When the inhaled oxygen fraction was 5% to 70%, R2* was linearly related to pO(2) levels (cortex DeltaR2*/DeltapO(2)=-1.2 ms(-1)kPa(-1), and medulla DeltaR2*/DeltapO2 =-1.7 ms(-1)kPa(-1)). Twenty-four hours of UUO was associated with an increased R2* in the cortex and a decreased R2* in medulla as compared with baseline, which remained augmented after the release of UUO, indicating that pO(2) levels were reduced in the cortex and increased in the medulla during and after release of obstruction. CONCLUSION: BOLD MRI provides noninvasive estimates of regional renal oxygen content and our study demonstrates that this technique may provide a useful tool in UUO which is associated with altered renal oxygen consumption. PMID- 15882273 TI - Urine protein patterns can serve as diagnostic tools in patients with IgA nephropathy. AB - BACKGROUND: IgA nephropathy (IgAN) is the most common chronic glomerular disease in adults. End-stage renal disease (ESRD) develops in about 30% of the patients. Early intervention and consequent therapy may prevent or at least delay the development of ESRD in these patients. Up to now, the diagnosis could only be achieved with a renal biopsy. METHODS: The urine of 45 patients with IgAN was collected and screened for protein/polypeptide patterns with a novel high throughput method, capillary electrophoreses on-line coupled to a mass spectrometer (CE-MS). CE-MS allows the fast and accurate evaluation of up to 2000 polypeptides in one urine sample. The results in IgAN were compared to findings in 13 patients with membranous nephropathy (MN) and 57 healthy individuals. RESULTS: In the patients with IgAN, even when urinary protein excretion was within the normal range of regular tests, the polypeptide pattern in urine differed significantly from that of healthy controls and patients with MN, indicating a specific "IgAN" pattern of polypeptide excretion. Classification regarding discrimination of IgAN from healthy controls and from MN had a sensitivity of 100% and 77%, respectively. Specificity was 90% and 100%, respectively. Compared to patterns established earlier in patients with minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), or diabetic nephropathy (DN), sensitivity and specificity were 100%. Treatment of the patients was associated with changes of the pattern, possibly indicating a therapeutic effect. CONCLUSION: Proteomic analysis with CE-MS coupling permits fast and accurate identification and differentiation of polypeptide patterns in the urine of patients with IgAN, allowing differentiation from healthy controls and, probably, other renal diseases. PMID- 15882274 TI - The Cockcroft-Gault formula should not be used in children. AB - BACKGROUND: Although designed for adults, the Cockcroft-Gault formula was recently proposed for use in children > or =13 years of age. METHODS: We compared the feasibility of the Cockcroft-Gault formula against the standard pediatric Schwartz formula and a novel cystatin C-based formula. Our patient cohort included 262 children aged 1 to 18 years with various renal pathologies, who underwent a 99-technetium diethylenetriaminepentaacetate ((99)Tc DTPA) glomerular filtration rate (GFR) renal scan. Calculations were performed in Systeme International (SI) units using published constants and recalculated constants from our patient population. Agreement was assessed using Bland and Altman analysis. RESULTS: Published and recalculated constants for the Cockcroft-Gault formula were 1.23 and 0.96, respectively, for boys, and 1.05 and 0.90, respectively, for girls. The published and recalculated constants for the Schwartz formula were 48 and 49.9, respectively, for boys > or =13 years old, and 38 and 46.2, respectively, for all girls and for boys <13 years old. Using published constants, there was agreement between GFR and Cockcroft-Gault formula in boys > or =13 years old (average bias 5.0 +/- 23.5%) while there was an average error of -19.0%+/- 36.4% for all ages. Similarly, the average bias with Schwartz for boys > or =13 years old was -6.8 +/- 24.0% and for all patients was 12.8 +/- 24.2%. Using recalculated constants, the average bias with Cockcroft Gault in boys > or =13 years old was -19.8 +/- 23.5% and for all patients was 38.5 +/- 35.2%. Similarly, the average bias with Schwartz for boys < or =13 years old was -1.1 +/- 24.3% and for all patients was 3.0 +/- 24.0%. The novel cystatin C-based GFR calculations showed an average error of -4.9 +/- 20.3% in the adolescent boys and 2.4 +/- 20.4% for all ages. CONCLUSION: Cockcroft-Gault formula showed the worst agreement with GFR, regardless of using published or recalculated constants. The cystatin C-based approach resulted in the least error, and should be used for estimation of GFR. PMID- 15882275 TI - Reduction of urinary connective tissue growth factor by Losartan in type 1 patients with diabetic nephropathy. AB - BACKGROUND: Connective tissue growth factor (CTGF) is an important profibrotic cytokine implicated in development of diabetic glomerulosclerosis. Urinary CTGF is reported to be significantly increased in patients with diabetic nephropathy. The present study aimed to investigate the short- and long term effects of angiotensin II receptor blockade by Losartan on urinary CTGF levels in hypertensive type 1 diabetic patients with diabetic nephropathy. METHODS: Seventy one hypertensive type 1 diabetic patients with diabetic nephropathy were included in the study. After a washout period of 4 weeks, the patients received Losartan 50 mg, 100 mg, and 150 mg once daily in treatment periods each lasting 2 months. Thereafter, patients were followed prospectively during treatment with Losartan 100 mg o.d. with a total mean follow-up time of 36 months. At baseline, after 2, 4, and 6 months and then biannually, urinary and plasma CTGF levels [enzyme linked immunosorbent assay (ELISA) fibroGen], albuminuria (Turbidimetry), glomerular filtration rate (GFR) [51-creatinine ethylenediaminetetraacetic acid ((51)Cr-EDTA plasma clearance)] and 24 hours blood pressure (TM2420)) were determined. RESULTS: Baseline levels of urinary and plasma CTGF were 7076 (5708 to 8770) ng/24 hours [geometric mean (95% CI)] and 12.7 (7.3) ng/mL [mean (SD)], respectively. Albuminuria, GFR, and arterial blood pressure at baseline were 1152 (937 to 1416) mg/24 hours, 88 (24) mL/min/1.73 m(2), and 153/80 (17/9) mm Hg, respectively. Losartan significantly reduced urinary CTGF by 21% (9 to 31) (95% CI) initially (P < 0.05 vs. baseline), with no further reduction after increasing dose. The sustained reduction in urinary CTGF was 22% (12 to 32) (P < 0.05 vs. baseline). Rate of decline in GFR during the study was 3.2 (-1.6 to 15.9) mL/min/year [median (range)]. Reduction in urinary CTGF was correlated with a lower rate of decline in GFR (r= 0.23, P= 0.05). Plasma CTGF remained unchanged throughout the study. CONCLUSION: Our 3-year study demonstrates that Losartan persistently reduces urinary CTGF excretion, which is associated with a slower rate of decline in GFR. PMID- 15882276 TI - Prognostic value of combined use of biomarkers of inflammation, endothelial dysfunction, and myocardiopathy in patients with ESRD. AB - BACKGROUND: Cardiovascular risk stratification is important in the clinical management of patients with end-stage renal diseases (ESRD) and biomarkers are increasingly used in these patients. METHODS: In a cohort of 246 dialysis patients without heart failure at baseline we tested the combined prognostic power of three well-established biomarkers: brain natriuretic peptide (BNP), C reactive protein (CRP), and asymmetric dimethyl arginine (ADMA). The independent prognostic value of individual and combined biomarkers was estimated in separate Cox models, including standard risk factors in dialysis patients and comorbidities. RESULTS: When the prediction power of the three biomarkers was evaluated individually, BNP, ADMA, and CRP added significant predictive value (P< or = 0.01) to all-cause and cardiovascular mortality models and the explanatory gain attributable to these biomarkers were of similar degree (ranging from 3.3% to 5.7%). When the biomarkers were evaluated jointly, a score based on the BNP CRP combination, increased by 9.9% (all-cause) and by 10.5% (cardiovascular) the explained mortality variance of standard Cox models and such gain in power was similar to that achieved by the CRP-ADMA combination (all-cause death 9.0% and cardiovascular death 8.4%). Of note, the explanatory gain derived by the simultaneous use of the three biomarkers was very similar (all-cause death 11.6% and cardiovascular death 10.5%) to that achieved by the use of two biomarkers. CONCLUSION: These findings indicate a potential role for CRP, BNP, and ADMA to be incorporated into diagnostic and therapeutic strategies aimed at detection and treatment of atherosclerotic complications and at preventing heart failure in the dialysis population. PMID- 15882277 TI - Parathyroid hormone and growth in children with chronic renal failure. AB - BACKGROUND: In pediatric chronic renal failure (CRF) optimal parathyroid hormone (PTH) concentrations that minimize renal osteodystrophy and maximize growth are unknown. The search for optimum concentrations has been complicated as currently used "intact" PTH (iPTH) assays cross-react with long carboxyl-terminal PTH fragments (C-PTH), which antagonize the biologic actions of 1-84 PTH. The purpose of this study was to investigate the relationship between PTH, the 1-84 PTH:C-PTH ratio and growth rate in children with CRF. METHODS: A total of 162 patients, median (range) age 9.9 years (0.3 to 17.1 years), were recruited: 136 with a glomerular filtration rate (GFR) <60 mL/min/1.73 m(2)[96 managed conservatively (CRF group) and 40 transplanted patients], and 26 dialysis patients. Over a median (range) period of 1.1 years (0.5 to 1.7 years), children attended five (three to 15) clinics at which iPTH, cyclase-activating PTH (CAP-PTH), and height were measured. RESULTS: Mean PTH concentrations were within the normal range for both assays for the CRF group and up to twice the upper limit of normal for the dialysis group; CAP-PTH 24.8 pg/mL and 59.9 pg/mL (normal range 5 to 39 pg/mL), iPTH 37.1 pg/mL, and 102.6 pg/mL, respectively (normal range 14 to 66 pg/mL). The patients grew normally (change in height standard deviation score per year (DeltaHtSDS) =-0.01). There was no relationship between PTH concentrations and DeltaHtSDS in any patient group. The 1-84 PTH:C-PTH ratio was lower in dialyzed patients (P= 0.003), with worsening renal function (P= 0.047) and with PTH concentrations outside the normal range (P= 0.01). There was a weak correlation between the 1-84 PTH:C-PTH ratio and the DeltaHtSDS (r= 0.2, P= 0.01). CONCLUSION: Normal range PTH concentrations are appropriate, allowing normal growth in children with CRF managed conservatively. C-PTH may be of clinical significance. PMID- 15882278 TI - The increased incidence of pure red cell aplasia with an Eprex formulation in uncoated rubber stopper syringes. AB - BACKGROUND: The incidence of pure red cell aplasia (PRCA) in chronic kidney disease patients treated with epoetins increased substantially in 1998, was shown to be antibody mediated, and was associated predominantly with subcutaneous administration of Eprex. A technical investigation identified organic compounds leached from uncoated rubber stoppers in prefilled syringes containing polysorbate 80 as the most probable cause of the increased immunogenicity. METHODS: This study investigated whether the incidence of PRCA was higher for exposure to the product form containing leachates than for leachate-free product forms. Antibody-mediated PRCA cases were classified according to indication, product form, and route of administration. Exposure estimates were obtained by country, indication, route of administration, and product form. RESULTS: For 2001 to 2003, the PRCA incidence rate for patients with subcutaneous exposure to Eprex in prefilled syringes with polysorbate 80 and uncoated rubber stoppers (leachates present) was 4.61/10,000 patient years (95% CI 3.88-5.43) versus 0.26/10,000 patient years (95% CI 0.007-1.44) for syringes with coated stoppers (leachates absent). The rate difference was 4.35/10,000 patient years (95% CI 3.44-5.26; P < 0.0001); the rate ratio was 17 (95% CI 3.14-707). A substantial rate difference remained in sensitivity analyses that adjusted for exposure to multiple product forms. CONCLUSION: The epidemiologic data, together with the chemical and immunologic data, support the hypothesis that leachates from uncoated rubber syringe stoppers caused the increased incidence of PRCA associated with Eprex. Currently, all Eprex prefilled syringes contain fluoro-resin coated stoppers, which has contributed to decreased incidence of PRCA with continued surveillance. PMID- 15882279 TI - Autosomal-dominant familial hematuria with retinal arteriolar tortuosity and contractures: a novel syndrome. AB - BACKGROUND: Autosomal-dominant forms of hematuria have been mostly related to mutations in the COL4A3/COL4A4 genes. Patients with thin basement membrane (BM) disease do not have extrarenal manifestations, while those with Alport syndrome often present with hearing loss, anterior lenticonus, and dot-and-fleck retinopathy. METHODS: We performed a phenotypic study and a candidate gene approach in a four-generation family presenting with autosomal-dominant hematuria associated with extrarenal manifestations. Renal biopsy was analyzed for determination of BM thickness and expression of chains of type IV collagen. Linkage to 18 candidate genes/loci was investigated using polymorphic microsatellite markers. RESULTS: In all affected patients, hematuria without proteinuria was associated with muscular contractures and retinal arterial tortuosities responsible for retinal hemorrhages. Cardiac arrhythmia, Raynaud phenomena, and brain MRI abnormalities were also observed. Despite the presence of red cells in tubule sections, no glomerular abnormalities were found by electron microscopy. Expression of type IV collagen chains and glomerular BM thickness was normal. We searched for a molecular defect affecting either BM or angiogenesis. Linkage analyses of genes encoding BM components (COL4A3/COL4A4, COL6A1, COL6A2, COL6A3, FBLN1), and angiogenic factors or their receptors (VHL, ANPT1, ANPT2, TIE, TEK, NOTCH2, NOTCH3, NOTCH4, DLL4, JAG1, JAG2) and of the facio-sapulo-humeral dystrophy and 3q21 loci failed to show segregation of the disease with those gene loci. CONCLUSION: We have identified a new inherited hematuria syndrome associated with retinal vessel tortuosities and contractures. We recommend performing a fundus examination in patients with familial hematuria and episodes of visual impairment, as well as a urinary analysis in patients with retinal arterial tortuosity or congenital muscular contractures. PMID- 15882280 TI - Regional citrate versus systemic heparin anticoagulation for continuous renal replacement in critically ill patients. AB - BACKGROUND: We determined the effect of regional citrate versus systemic heparin anticoagulation for continuous renal replacement therapy in critically ill subjects suffering from acute renal failure who were not at high risk for hemorrhagic complications. METHODS: Between April 1999 and June 2002, 30 critically ill subjects requiring continuous renal replacement therapy and using 79 hemofilters were randomly assigned to receive regional citrate or systemic heparin anticoagulation. RESULTS: The median hemofilter survival time was 124.5 hours (95% CI 95.3 to 157.4) in the citrate group, which was significantly longer than the 38.3 hours (95% CI 24.8 to 61.9) in the heparin group (P < 0.001). Increasing illness severity score, male gender, and decreasing antithrombin-III levels were independent predictors of an increased relative hazard of hemofilter failure. After adjustment for illness severity, antithrombin-III levels increased significantly more over the period of study in the citrate as compared to the heparin group (P= 0.038). Moreover, after adjustment for antithrombin-III levels and illness severity score, the relative risk of hemorrhage with citrate anticoagulation was significantly lower than that with heparin (relative risk of 0.14; 95% CI 0.02 to 0.96, P= 0.05). CONCLUSION: Compared with systemic heparin anticoagulation, regional citrate anticoagulation significantly increases hemofilter survival time, and significantly decreases bleeding risk in critically ill patients suffering from acute renal failure and requiring continuous renal replacement therapy. PMID- 15882281 TI - The effect of a first kidney transplant on a subsequent transplant outcome: an experimental and clinical study. AB - BACKGROUND: Second kidney transplantations have a roughly similar clinical outcome to first transplantations. Nevertheless, the effect of the presence of the first, nonfunctional transplant at the time of the second transplantation may also influence its outcome and has not yet been specifically studied. METHODS: We analyzed the effect of the presence of a first graft on the outcome of a second graft in a rodent allograft model and in a cohort of 240 human second kidney allograft recipients. RESULTS: In rodents, 100 days subsequent to the rejection of the first graft, we observed an increase in blood but not spleen CD4(+)CD25(+) T cells, whereas no differences were observed in transcriptional patterns. Adoptive transfer of day 100 splenocytes did not prolong graft survival. Moreover, the presence of a first rejected graft does not prolong the survival of a second graft performed at a later date. In the human context, a higher incidence of patients with anti-HLA immunization and a higher % of PRA were observed in retransplant recipients with primary allograft nephrectomy. Despite a relatively low statistical power, our data do not suggest significant differences in graft outcome between recipients of second transplants with primary allograft nephrectomy and those without. CONCLUSION: Collectively, the data from both an experimental model and a large cohort of human recipients of a second graft do not suggest a beneficial effect of the presence of a first rejected graft at the time of a second transplantation. PMID- 15882282 TI - The association between benign prostatic hyperplasia and chronic kidney disease in community-dwelling men. AB - BACKGROUND: Benign prostatic hyperplasia (BPH) and chronic kidney disease are important public health problems in older men. Previous referral-based studies disagree on whether BPH is associated with chronic kidney disease. The objective of this study was to determine the community-based association between clinical measures of BPH and chronic kidney disease. METHODS: A community-based sample of 2115 white men (ages 40-79 years) was randomly selected from the Olmsted County, Minnesota population (55% participation rate) in 1990. A random subsample (N= 476) had a detailed clinical evaluation. This evaluation included a questionnaire with similar queries to the International Prostate Symptom Score (IPSS), peak urinary flow rates (uroflowmeter), postvoid residual urine volume (ultrasound), prostate volume (ultrasound), serum prostate specific antigen (PSA), and serum creatinine. RESULTS: After adjustment for age, hypertension, diabetes, leukocyte esterase positive (possible urinary tract infection), and smoking, chronic kidney disease [serum creatinine > or =133 micromol/L (1.5 mg/dL)] was associated with diminished peak urinary flow rate (<15 mL/sec) by an odds ratio (OR) = 2.96 (95% CI 1.30-7.01), moderate-severe lower urinary tract symptoms (IPSS >7) by an OR = 2.91 (95% CI 1.32-6.62), and chronic urinary retention (postvoid residual >100 mL) by an OR = 2.28 (95% CI 0.66-6.68). There was no association with a prostate volume >30 mL by an OR = 0.56 (95% CI 0.22-1.37) or PSA >1.4 ng/mL by an OR = 1.17 (95% CI 0.47-2.81). CONCLUSION: There was a cross-sectional association between signs and symptoms of bladder outlet obstruction and chronic kidney disease in community-dwelling men. Prostatic enlargement was not associated with chronic kidney disease. PMID- 15882283 TI - Low fetuin-A levels are associated with cardiovascular death: Impact of variations in the gene encoding fetuin. AB - BACKGROUND: Vascular calcification is common among end-stage renal disease (ESRD) patients and a central characteristic of the atherosclerotic cardiovascular disease observed in dialysis patients. Fetuin-A, a circulating calcium-regulatory glycoprotein that inhibits vascular calcification, is associated with inflammation and outcome in dialysis patients. In the present study, we evaluated the association between fetuin-A, clinical phenotype, and outcome, as well as the impact of fetuin gene (AHSG) polymorphisms on the protein product and outcome. METHODS: In a cohort of 258 (161 males) ESRD patients starting renal replacement therapy [glomerular filtration rate (GFR) 6.8 +/- 0.2 mL/min] aged 52 +/- 1 years the following parameters were studied: presence of malnutrition (subjective global assessment), comorbidity [diabetes mellitus and clinical manifest cardiovascular disease (CVD)], carotid plaques (N= 101), hs-CRP, fetuin-A, S albumin, interleukin (IL)-6, and single nucleotide polymorphisms (SNPs) in the AHSG gene (N= 215) at amino acid positions Thr248Met (C-->T), Thr256Ser (C-->G), Asp276Asn (G-->A), and Arg317Cys (C-->T). RESULTS: Both all-cause (P < 0.001) and cardiovascular (P < 0.001) mortality were associated with low fetuin-A levels independently of age, smoking, diabetes, S-albumin, CVD, and inflammation (CRP > or =10 mg/L). Inflamed (0.199 vs. 0.247 g/L; P < 0.01) and malnourished (0.207 vs. 0.262 g/L; P < 0.05) patients had significantly lower median fetuin-A than noninflamed and well-nourished ESRD patients, respectively. In a logistic regression model (N= 101), fetuin-A was significantly (P < 0.05) associated with the presence of carotid plaques independently of age, CVD, diabetes, S-albumin, gender, and inflammation. Significant correlations were observed between fetuin-A and both S-albumin (Rho = 0.30; P < 0.0001) and IL-6 (Rho =-0.21; P < 0.01). Patients with the AHSG 256Ser allele had lower serum fetuin-A levels, and higher all-cause and cardiovascular mortality rate if they were inflamed. CONCLUSION: The present study shows that a low fetuin-A level is associated with malnutrition, inflammation, and atherosclerosis (carotid plaques), as well as with increased cardiovascular and all-cause mortality. Because the present study demonstrates an effect of variations in the AHSG gene on both circulating fetuin A levels and outcome, this indicates that ESRD patients with the AHSG 256Ser allele are at risk of accelerated vascular calcification. PMID- 15882284 TI - Case-control study of regular analgesic and nonsteroidal anti-inflammatory use and end-stage renal disease. AB - BACKGROUND: Studies on the association between the long-term use of aspirin and other analgesic and nonsteroidal anti-inflammatory drugs (NSAIDs) and end-stage renal disease (ESRD) have given conflicting results. In order to examine this association, a case-control study with incident cases of ESRD was carried out. METHODS: The cases were all patients entering the local dialysis program because of ESRD in the study area between June 1, 1995 and November 30, 1997. They were classified according to the underlying disease, which had presumably led them to ESRD. Controls were patients admitted to the same hospitals from where the cases arose, also matched by age and sex. Odds ratios were calculated using a conditional logistic model, including potential confounding factors, both for the whole study population and for the various underlying diseases. RESULTS: Five hundred and eighty-three cases and 1190 controls were included in the analysis. Long-term use of any analgesic was associated with an overall odds ratio of 1.22 (95% CI, 0.89-1.66). For specific groups of drugs, the risks were 1.56 (1.05 2.30) for aspirin, 1.03 (0.60-1.76) for pyrazolones, 0.80 (0.39-1.63) for paracetamol, and 0.94 (0.57-1.56) for nonaspirin NSAIDs. The risk of ESRD associated with aspirin was related to the cumulated dose and duration of use, and it was particularly high among the subset of patients with vascular nephropathy as underlying disease [2.35 (1.17-4.72)]. CONCLUSION: Our data indicate that long-term use of nonaspirin analgesic drugs and NSAIDs is not associated with an increased risk of ESRD. However, the chronic use of aspirin may increase the risk of ESRD. PMID- 15882285 TI - Conversion of tunneled hemodialysis catheter-consigned patients to arteriovenous fistula. AB - OBJECTIVE: Despite their high incidence of complications, costs, morbidity, and mortality, nearly 27% of the chronic hemodialysis (HD) patients are receiving treatment via a tunneled hemodialysis catheter (TDC). METHODS: In this prospective analysis, an interventional nephrology team employed an organized program consisting of vascular access (VA) education and vascular mapping (VM) to TDC-consigned patients. A full range of surgical approaches for arteriovenous fistula (AVF) creation, including vein transpositions, was exercised. Physical examination was performed every 1 to 2 weeks after surgery to assess the development of the AVF. Fistulas that failed to develop adequately to support HD (early failure) underwent salvage [percutaneous transluminal angioplasty (PTA), accessory vein obliteration (AVL)] procedures. RESULTS: One hundred twenty-one TDC-consigned patients received VA education. Eighty-six (71%) agreed to undergo VM. Two groups were identified. Group I (N= 66; using TDC for 7.2 +/- 1.8 SD months) had never had an arteriovenous access; group II (N= 20; using TDC for 12.3 +/- 4.0 months) had a history of one or more previously failed arteriovenous accesses. Upon VM, 64/66 (97%) in group I and 18/20 (90%) in group II were found to have adequate veins for AVF creation. Seven patients (11%) in group I and 3 (17%) in group II refused surgery. In group I, 57 (89%) received an arteriovenous access (radiocephalic AVF = 15, brachiocephalic AVF = 35, transposed brachiobasilic AVF = 3, brachiobasilic AVG = 4). In group II, 15 (83%) received a transposed AVF (radiobasilic = 2, brachiobasilic = 13). Sixteen fistulas (30%) in group I and 8 (53%) in group II had early failure. All except for one fistula in each group were salvaged using PTA and/or AVL. All 70 accesses (AVF = 66, AVG = 4) remain functional, with a mean follow-up of 8.5 +/- 3.6 months. CONCLUSION: These results demonstrate that an organized approach based upon a comprehensive program utilizing VA counseling, VM, application of full range of surgical techniques, and salvage procedures can be very successful in providing optimum vascular access to the catheter-dependent patient. PMID- 15882286 TI - The advanced age deceased kidney donor: current outcomes and future opportunities. AB - BACKGROUND: Due to the aging general population, deceased donors > or =55 years will form an increasingly larger proportion of the deceased kidney donor pool. METHODS: Using data from the United States Renal Data System, we determined the change in graft survival between 1996 and 2000 among 32,557 recipients of donors aged <55 years and > or =55 years in univariate and multivariate survival analyses. We identified donor risk factors for graft loss that might influence the decision to accept or reject donors <55 and > or =55 years. The initial glomerular filtration rate established 6 months after transplantation (initial GFR), and the stability of GFR in the first post-transplant year (GFR at 12 months post-transplantation-GFR at six months post-transplantation) were compared between recipients of donors <55 and > or =55 years and the association of these factors with graft survival was determined. RESULTS: In 2000, one-year graft survival in donors > or =55 years was 86.7%. Between 1996 and 1999 the projected graft half life improved from 11.4 to 14.5 years for recipients of donors <55 years (P < 0.01); however, there was no improvement for recipients of donors > or =55 years (8.2 to 9.2 year, P= 0.46). Among donor factors studied, only cold ischemic time >24 hours identified recipients of donors > or =55 years at risk for graft loss. Compared to recipients of donors <55 years, recipients of donors > or =55 years established a lower initial GFR (42 vs. 56 mL/min/1.73 m(2), P < 0.0001), and had less stable GFR in the first post-transplant year (-1.5 vs. -0.6 mL/min/1.73 m(2), P <.0001). Recipients from donors > or =55 years with initial GFR > or =50 mL/min/1.73 m(2) and no drop GFR during the first post-transplant year had graft survival that was superior to that of donors <55 years with either initial GFR <50 mL/min/1.73 m(2) or a drop in GFR during the first post transplant year. CONCLUSION: Donors > or =55 years are a valuable resource. Despite improvements in immunosuppression, rejection, and delayed graft function, the projected increase in long-term graft survival among recipients of donors <55 years was not shared among recipients of donors > or =55 years. Recipients of donors > or =55 years had lower initial GFR, and less stable GFR during the first post-transplant year. Limiting cold ischemic time to <24 hours may improve outcomes among recipients of donors > or =55 years. Future studies to maximize initial GFR and minimize early loss of GFR in recipients of donors > or =55 years may lead to improved outcomes from deceased donors > or =55 years. PMID- 15882287 TI - New onset hyperglycemia and diabetes are associated with increased cardiovascular risk after kidney transplantation. AB - BACKGROUND: Post-transplant diabetes (PTDM) is a common and serious complication of kidney transplantation. The implications of developing hyperglycemia of lesser severity are not well understood. METHODS: In this study we used American Diabetes Association (ADA) criteria to assess the incidence of abnormal glycemia post-transplant, the variables that relate to this complication, and the relationship between hyperglycemia and cardiovascular (CV) disease. Included in the study were 490 kidney recipients, transplanted from 1998 to 2003, without a history of diabetes, and with a pretransplant fasting glucose <126 mg/dL. RESULTS: Within one week post-transplant, 45% of recipients had impaired fasting glycemia (IFG, glucose 100-125 mg/dL), and 21% PTDM (glucose > or =126). One year post-transplant, 33% of patients had IFG, and 13% PTDM. Risk factors for hyperglycemia at one year included: older recipient, male gender, higher BMI, higher pretransplant glucose, and higher glucose one week post-transplant (all P < 0.002 by multivariable analyses). During a follow-up period of 40 +/- 14 months, 12% of recipients had CV events (cardiac, CVA, and/or peripheral). Increasing fasting glucose levels at one, four, and/or 12 months post-transplant were significantly related to CV events. Furthermore, these relationships were independent of other CV risk factors, including: older age, CV events pretransplant, male gender, dyslipidemia, and transplant year. Fasting glucose levels >100 mg/dL were associated with higher incidence of post-transplant cardiac (P= 0.001) and peripheral vascular disease events (P= 0.003). CONCLUSION: The incidence of post-transplant hyperglycemia and its CV impact have been underestimated. Pretransplant characteristics and, particularly, the glycemia during the first month post-transplant identified patients at risk of PTDM. Increasing glucose levels greater than 100 mg/dL, any time after the first month post-transplant, are associated with increasing CV risk. We postulate that aggressive detection and treatment of post-transplant hyperglycemia may significantly reduce CV morbidity and mortality after kidney transplantation. PMID- 15882288 TI - Rapamycin inhibits human renal epithelial cell proliferation: effect on cyclin D3 mRNA expression and stability. AB - BACKGROUND: Recent data have suggested that rapamycin use during the initial period after transplantation is associated with prolonged delayed graft function (DGF). Because of the known effects of rapamycin in other cell types, we speculated that this action may be secondary to human renal epithelial cells (HRECs) inhibition of proliferation. METHODS: Primary cultures of HRECs were incubated with various concentrations of rapamycin. Cell proliferation was evaluated by cytotoxicity assays. The cell cycle was analyzed by flow cytometry. Protein expression levels were assessed by Western blot. Cyclin D3 mRNA levels were measured by quantitative real-time polymerase chain reaction (PCR). The transcriptional activity of the cyclin D3 gene was evaluated using transient transfection. RESULTS: Rapamycin exerted a significant concentration-dependent antiproliferative effect on growing HRECs by inhibiting the G(1) to S transition. The p70(S6) kinase pathway leading to cell cycle progression was found to be active, and low concentrations of rapamycin dramatically reduced p70(S6) kinase phosphorylation. Rapamycin completely inhibited the increase in cyclin D3 protein expression and mRNA accumulation induced by fetal calf serum, but did not affect cyclin E or cdk-inhibitor expression levels. This regulation of cyclin D3 protein expression is mainly due to a destabilization of its mRNA. Rapamycin reduced the mRNA half-life by 26% (4.8 +/- 1.3 hours vs. 6.5 +/- 1.0 hours, P < 0.001). CONCLUSION: Rapamycin inhibits the proliferative response of HRECs to mitogenic stimuli, and causes cell cycle arrest in the early G(1) phase, not only by a nonspecific process due to inhibition of the p70(S6k) pathway, but also by a direct effect on cyclin D3 mRNA stability. PMID- 15882289 TI - Silent cerebral infarction predicts vascular events in hemodialysis patients. AB - BACKGROUND: Cardiovascular disease is the leading cause of death in hemodialysis (HD) patients. We have previously reported a higher incidence of silent cerebral infarction (SCI) in HD patients compared with the control group using MRI studies. In the present study, we examined whether or not SCI could predict vascular events in HD patients. METHODS: Cranial magnetic resonance imaging (MRI) was performed on 119 HD patients without symptomatic cerebrovascular disease. SCI was detected by MRI, and the patients were prospectively followed up. The end points of the study were the incidence of major events related to vascular events (cerebral events, cardiac events, and sudden deaths). We investigated the prognostic role of SCI in cerebral, cardiac, and vascular events by using the Kaplan-Meier method and Cox proportional hazards analysis. RESULTS: The prevalence of SCI was 49.6% in HD patients. During a follow-up period of maximum 60 months, vascular events, which included 13 cerebral events, 5 cardiac events, and 3 sudden deaths, occurred in 21 patients. The presence of SCI was predictive for a higher cerebral and vascular morbidity compared to the absence of SCI [18.6% (N= 11) vs. 3.3% (N= 2), P= 0.0169, and 30.5% (N= 18) vs. 5.0% (N= 3), P= 0.0006, respectively]. By multivariate Cox proportional hazards analysis, SCI remained a powerful independent predictor of cerebral and vascular events (hazard ratio for cerebral events 7.33, 95% CI 1.27-42.25: for vascular events 4.48, 95% CI 1.09-18.41). CONCLUSION: The findings of the present study indicate that the presence of SCI is a new risk factor for vascular events in HD patients. PMID- 15882290 TI - AUC-guided dosing of tacrolimus prevents progressive systemic overexposure in renal transplant recipients. AB - BACKGROUND: Tacrolimus has a narrow therapeutic window, and bioavailability is known to vary considerably between renal transplant recipients. Most centers still rely on measurement of trough levels, but there are conflicting reports on the correlation between tacrolimus trough levels and systemic exposure, as measured by the area-under-the-concentration-over-time curve (AUC((0-12h))). METHODS: We developed and validated a two-compartmental population-based pharmacokinetic model with Bayesian estimation of tacrolimus systemic exposure. Subsequently, we used this model to apply prospectively AUC-guided dosing of tacrolimus in 15 consecutive renal transplant recipients. The main objective was to study intrapatient variability in the course of time. RESULTS: Bayesian forecasting with a two-point sampling strategy, a trough level, and a second sample obtained between two and four hours post-dose significantly improved the squared correlation with the AUC((0-12h)) (r(2)= 0.94). Compared with trough level monitoring only, this approach reduced the 95%-prediction interval by 50%. The Bayesian approach proved to be feasible in clinical practice, and provided accurate information about systemic tacrolimus exposure in individual patients. In the AUC-guided dosing cohort the apparent clearance of tacrolimus decreased gradually over time, which was not reflected in corresponding trough levels. CONCLUSION: This simple, flexible method provides the opportunity to tailor immunosuppression, and should help minimize tacrolimus-related toxicity, such as nephrotoxicity and post-transplant diabetes mellitus. PMID- 15882291 TI - Access to, and outcome of, renal transplantation according to treatment modality of end-stage renal disease in France. AB - BACKGROUND: Although peritoneal dialysis (PD) is recognized as one of the methods of treatment of end-stage renal disease (ESRD), there have been recurrent concerns about the access of patients treated by this modality to kidney transplantation (KTx), as well as reports showing increased complications of KTx in such patients, such as graft thrombosis and infections. METHODS: The aim of this study was to provide a comprehensive view of the impact on transplantation of pretransplant modality of treatment of ESRD using a multivariate analysis of the French database. From 1997 to 2000, after exclusion of pediatric patients, multiple transplantations, and living donors, 6420 were patients registered on the waiting list, and 3464 were transplanted. RESULTS: Using a Cox proportional hazard analysis, we found a shorter waiting time for PD patients (RR 0.71, P < 0.0001), which became equivalent to hemodialysis (HD) patients when taking into account the transplant center as a variable (RR 1.0, P= 0.95). Concerning graft survival, only preemptive transplantation had a significant impact, being associated to a decreased risk of graft failure (RR 0.46, P= 0.005). Conclusion. Our study supports the concept that the choice of any pretransplant dialysis modality does not influence waiting time for transplant or the results of transplantation. PMID- 15882292 TI - Relevance of Toll-like receptor-4 polymorphisms in renal transplantation. AB - BACKGROUND: Polymorphisms in Toll-like receptor-4 (TLR4) have been reported to be associated with a blunted immune response to microbial pathogens, as well as a decreased risk of atherosclerosis in the general population. We assessed the impact of the two TLR4 variants on the risk of severe infection, the incidence of acute rejection, and the occurrence of atherosclerotic complications in renal transplant recipients (RTR). METHODS: TLR-4 polymorphisms were assessed in a cohort of 238 RTR. Post-transplant atherosclerotic events, acute rejection, severe bacterial infection, cytomegalovirus (CMV) disease, and opportunistic infections were evaluated as outcomes. RESULTS: The patients were followed for a mean duration of 95 +/- 29 months after transplant. TLR4 polymorphism was observed in 27 (11.3%) RTR. Subjects with TLR4 polymorphisms were less likely to experience post-transplant atherosclerotic events (RR 0.44; 95% CI 0.21 to 0.93; P= 0.02) and acute rejection (RR 0.41; 95% CI 0.30 to 0.83; P= 0.01), but presented severe bacterial infections (RR 1.33; 95% CI 1.12 to 1.67; P= 0.01) and opportunistic infections (RR 3.03; 95% CI 1.72 to 8.29; P= 0.008) more frequently. TLR4 polymorphism was marginally associated with CMV disease (RR 1.47; 95% CI 0.95 to 2.64; P= 0.08). CONCLUSION: RTR with TLR4 polymorphism present a lower risk of post-transplant atherosclerotic events and acute allograft rejection, but experience severe infectious episodes more frequently. This subset of RTR may benefit from a less potent immunosuppression regimen, along with increased preventive measures against infectious agents. PMID- 15882293 TI - Arteriovenous fistula outcomes in the era of the elderly dialysis population. AB - BACKGROUND: The growth of patients > or =65 years on hemodialysis is increasing. Guidelines recommend arteriovenous fistula (AVF) access but their outcomes in elderly patients are controversial. This study compared the outcomes of AVF in patients <65 years old (65- group) versus those > or =65 years old (65+ group). METHODS: This retrospective analysis of prospectively collected data included 444 incident, first-time AVF created in a large dialysis center between January 1, 1995 and July 1, 2003. The primary outcome of AVF cumulative patency was evaluated using Kaplan-Meier survival analysis with log-rank test comparison. A Cox model determined factors associated with AVF loss. RESULTS: One hundred ninety-six patients (44%) were in the 65+ group. In total, there were 230 (52.2%) radiocephalic, 186 (42.2%) brachiocephalic, and 25 (5.6%) basilic vein transposed AVF. The one-year AVF cumulative survival was 75.1% (65+ group) and 79.7% (65- group); the five-year survival was 64.7% (65+ group) and 71.4% (65- group). The overall total procedure, angioplasty, thrombolysis, and revision rates per access year were 0.83, 0.30, 0.66, and 0.16, respectively. The 65+ group had a relative risk of 1.7 of their AVF failing to mature compared with the 65- group. Multivariate analysis yielded these variables significant for AVF loss: male sex HR 0.63 (95% CI 0.44-0.91), coronary artery disease HR 2.1 (95% CI 1.5-3.0), and Caucasian ethnicity HR 0.63 (95% CI 0.44-0.91). CONCLUSION: Age should not be a limiting factor when determining candidacy for AVF creation due to equivalent survival and procedural rates. Failure of fistula maturation is a primary concern to patients of all ages and demands further study. PMID- 15882294 TI - Dialysate partitioning in the Genius batch hemodialysis system: effect of temperature and solute concentration. AB - BACKGROUND: The Genius batch system contains a 75-L closed reservoir from which fresh dialysate is extracted at the top, and to which spent dialysate is returned at the bottom. In vivo studies have demonstrated that almost the entire amount of dialysate can be used before contamination of fresh with spent dialysate occurs. The question is raised whether density differences cause this separation, and what the relative contributions of temperature and solute content are. METHODS: As patient substitute, a container filled with dialysate was loaded with various amounts of urea. Temperature differences between spent and fresh dialysate were imposed by not heating the dialysate at the outlet line from the dialyzer (A), heating the outlet to obtain continuously equal temperatures at inlet and outlet (B), or to temperatures as in vivo (C). With a dialysate flow set at 300 mL/min, urea is not expected at the inlet before 250 minutes. RESULTS: With a urea concentration of 33 mg/dL, urea contamination at the dialysate inlet line occurred after 185 +/- 20 (A), 122 +/- 11 (B), and 175 +/- 12 minutes (C) of dialysis, whereas with 67 mg/dL, this happened at 219 +/- 5 (A), 162 +/- 11 (B), and 202 +/- 8 minutes (C). With 100 and 150 mg/dL, urea contamination appeared at 224 +/- 2 (A) and 204 +/- 14 minutes (B), and 227 +/- 5 (A) and 232 +/- 3 minutes (B), respectively. CONCLUSION: Both temperature differences between spent and fresh dialysate and solute content of spent dialysate contribute to dialysate partitioning in the Genius dialysis system. PMID- 15882295 TI - Genetic and clinical factors influence the baseline permeability of the peritoneal membrane. AB - BACKGROUND: Patients starting peritoneal dialysis (PD) show a significant variability in small solute transport across the peritoneal membrane (PM). The latter parameter determines dialysis prescription and survival. Clinical factors probably influence solute transport across the PM, but the putative role of genetic variants is unknown. METHODS: We have investigated the influence of functional polymorphisms of VEGF, ENOS, and IL-6, together with clinical and biological factors, on baseline peritoneal equilibration test (PET) parameters in a homogeneous population of 152 unrelated Caucasian PD patients from Belgium and the North of France. RESULTS: The distribution of the 21 alleles (7 polymorphisms) and linkage disequilibrium parameters were similar in PD patients and healthy subjects. Univariate and multivariate analyses identified comorbidity, serum albumin, and the -174G/C polymorphism of IL-6 as independent predictors of small solute transport. The -174G/C polymorphism of IL-6 was associated with significantly higher IL-6 mRNA levels in the PM and higher plasma and dialysate IL-6 concentrations, suggesting a dominant effect of the C allele. Patients harboring the CC and GC genotypes (N= 92) were characterized by significantly higher permeability parameters and inflammatory markers than patients harboring the GG genotype (N= 60). In contrast with IL-6, VEGF and ENOS polymorphisms had no influence on baseline peritoneal permeability. CONCLUSION: These data (1) show that, together with clinical parameters, the functionally relevant -174G/C polymorphism of IL-6 contributes to the interpatient variability in small solute transport rate at the start of PD; and (2) substantiate the critical role played by IL-6 in the PM. PMID- 15882296 TI - Antibodies against macrophages that overlap in specificity with fibroblasts. AB - BACKGROUND: Fibroblasts can be misidentified as macrophages because both cell types share antigens that are associated with popular antibodies targeting the monocyte/macrophage lineage. With the recent description of fibroblast-specific protein 1 (FSP1), we revisited the specificity of antibodies directed against macrophages to determine systematically which antibodies best distinguish both cell types in fibrotic tissues. METHODS: Tissue fibrosis was produced in mice carrying the GFP transgene encoding green fluorescent protein under the control of the FSP1 promoter. Single cell suspensions from these marked tissues were submitted for flow cytometry using antibodies against Mac-1, Mac-2, Mac-3, F4/80, CD68, major histocompatibility complex (MHC) class II, and CD45, and cDNA amplification of mRNA encoding the above target antigens was performed using specific primer sets in sorted pools of cells. Fibrotic tissues were also stained by immunohistochemistry with the same antibodies and examined under confocal microscopy. RESULTS: Comparison overlap between FSP1(+) fibroblasts with each of the macrophage markers demonstrated that all antimacrophage antibodies (Mac-1, Mac-2, Mac-3, CD68, MHC class II, and CD45) except one (F4/80) recognize both cell types. CONCLUSION: Antibodies directed against F4/80 clearly distinguish macrophages from FSP1(+) fibroblasts in fibrotic tissues and is the preferred antibody in mice. PMID- 15882297 TI - Increased tubular organic ion clearance following chronic ACE inhibition in patients with type 1 diabetes. AB - BACKGROUND: The tubular excretion of creatinine significantly contributes to its clearance. Administration of an angtiotensin-converting enzyme (ACE) inhibitor is associated with increased organic ion clearance in experimental diabetes. This study examines the effect and implications of chronic ACE inhibition on renal organic ion excretion in patients with type 1 diabetes. METHODS: Samples were obtained from the Melbourne Diabetic Nephropathy Study Group (MDNSG) that randomized patients to receive perindopril (N= 11), nifedipine (N= 11), or placebo (N= 8). Albumin excretion rate, creatinine clearance, and isotopic glomerular filtration rate (GFR) were assessed at baseline and after 24 months. In addition, the clearance of the endogenous cations N-methylynicotinamide (NMN), creatinine, and the anion hippurate were determined by high-performance liquid chromatography (HPLC). RESULTS: Following treatment with the ACE inhibitor, perindopril, renal clearance of NMN was increased (+96%) (P < 0.05). There was no difference in patients treated with nifedipine (P= 0.25) and NMN clearance fell in the placebo-treated patients (-26%) (P < 0.05). Changes in NMN clearance were unaffected after adjusting for the effects of perindopril on GFR. However, they were attenuated after adjusting for hippurate clearance, a marker of renal blood flow. This effect of perindopril on NMN clearance was seen in both men and women, regardless of baseline clearance and was correlated with reduced albuminuria following perindopril treatment. CONCLUSION: Organic ion clearance is increased in patients with diabetes following chronic ACE inhibition. This is consistent with experimental models showing increased ion transporter expression and improved tubular blood flow, following blockade of the renin-angiotensin system (RAS). These findings may have implications for the interpretation of creatinine based indices in patients with diabetes. PMID- 15882298 TI - HGF and extracellular matrix turnover in normal human glomeruli ex vivo. PMID- 15882299 TI - An unexpected role of plasminogen activator inhibitor-type 1 (PAI-1) in renal fibrosis. PMID- 15882301 TI - Neutrophil activation and renal scarring. PMID- 15882302 TI - Locking hemodialysis catheters with cefotaxime instead of gentamicin to avoid potential ototoxicity. PMID- 15882303 TI - Pharmacologic intervention to prevent hemodialysis vascular access thrombosis: the next generation of treatment? PMID- 15882304 TI - Prevention of renal cell carcinoma from hemodialysis patients by regulating epigenetic factors. PMID- 15882305 TI - The differential impact of risk factors on mortality in hemodialysis and peritoneal dialysis. PMID- 15882306 TI - Bacterial infections in hemodialysis patients: pathogenesis and prevention. PMID- 15882307 TI - Consequences and management of hyperphosphatemia in patients with renal insufficiency. AB - Consequences and management of hyperphosphatemia in patients with renal insufficiency. Progressive renal insufficiency leads to hyperphosphatemia, hypocalcemia, and secondary hyperparathyroidism. Bone demineralization in secondary hyperparathyroidism may induce fractures, while joint and subcutaneous precipitations of calcium pyrophosphate limit mobility, and may cause crippling. Strategies to preempt bone and joint destruction in chronic kidney disease and end-stage renal disease have focused on limiting dietary phosphorus, intra-gut binding of ingested phosphorous, enhancing calcium absorption, and limiting parathyroid hormone secretion. Deciding which regimen is most effective to meet these treatment objectives challenges nephrologists; they often uncover conflicting evidence about which abnormal metabolite should be the prime treatment objective. Especially vexing is the question of whether hypercalcemia is a cardiotoxic consequence of calcium-based phosphate binders. PMID- 15882308 TI - Treatment of hyperphosphatemia in patients with chronic kidney disease on maintenance hemodialysis. AB - Treatment of hyperphosphatemia in patients with chronic kidney disease on maintenance hemodialysis. Hyperphosphatemia in patients with ESRD leads to secondary hyperparathyroidism, renal osteodystrophy, and is independently associated with mortality risk. The exact mechanism by which hyperphosphatemia increases mortality risk is unknown, but it may relate to enhanced cardiovascular calcification. National Kidney Foundation K/DOQI bone metabolism and disease guidelines recommend maintenance of serum phosphorus (P) below 5.5 mg/dL, and Ca x P product less than 55 mg(2)/dL(2). Although calcium-based phosphate binders (CBPB) are cost effective, long-term safety concerns relate to their postulated role in progression of cardiovascular calcification. Sevelamer hydrochloride has been recommended as an alternative noncalcium phosphate binder. Results from the Calcium Acetate Renagel Evaluation (CARE study) indicate that calcium acetate is more effective than sevelamer in controlling serum phosphorous and Ca x P product in hemodialysis patients. In the Treat-to-Goal study, dialysis patients treated with sevelamer had slower progression of coronary and aortic calcification than patients treated with CBPB. The mechanism underlying the beneficial effect of sevelamer is unknown, but may relate to decreased calcium loading or to dramatic reductions in LDL cholesterol in sevelamer-treated patients. At present, evidence incriminating CBPB in the progression of cardiovascular calcification in ESRD remains largely circumstantial. As calcium acetate is more efficacious and cost effective than sevelamer, it remains an accepted first-line phosphate binder. In this review, we will examine these issues and provide rational guidelines for the use of calcium-based phosphate binders in patients on maintenance hemodialysis. PMID- 15882309 TI - Risks of chronic metabolic acidosis in patients with chronic kidney disease. AB - Risks of chronic metabolic acidosis in patients with chronic kidney disease. Metabolic acidosis is associated with chronic renal failure (CRF). Often, maintenance dialysis therapies are not able to reverse this condition. The major systemic consequences of chronic metabolic acidosis are increased protein catabolism, decreased protein synthesis, and a negative protein balance that improves after bicarbonate supplementation. Metabolic acidosis also induces insulin resistance and a decrease in the elevated serum leptin levels associated with CRF. These three factors may promote protein catabolism in maintenance dialysis patients. Available data suggest that metabolic acidosis is both catabolic and anti-anabolic. Several clinical studies have shown that correction of metabolic acidosis in maintenance dialysis patients is associated with modest improvements in nutritional status. Preliminary evidence indicates that metabolic acidosis may play a role in beta2-microglobulin accumulation, as well as the hypertriglyceridemia seen in renal failure. Interventional studies for metabolic acidosis have yielded inconsistent results in CRF and maintenance hemodialysis patients. In chronic peritoneal dialysis patients, the mitigation of acidemia appears more consistently to improve nutritional status and reduce hospitalizations. Large-scale, prospective, randomized interventional studies are needed to ascertain the potential benefits of correcting acidemia in maintenance hemodialysis patients. To avoid adverse events, an aggressive management approach is necessary to correct metabolic acidosis. Clinicians should attempt to adhere to the National Kidney Foundation Kidney Disease Outcome Quality Initiative (K/DOQI) guidelines for maintenance dialysis patients. The guidelines recommend maintenance of serum bicarbonate levels at 22 mEq/L or greater. PMID- 15882310 TI - The role of daily dialysis in the control of hyperphosphatemia. AB - The role of daily dialysis in the control of hyperphosphatemia. In patients with end-stage renal disease (ESRD), hyperphosphatemia occurs in the vast majority of patients. The numerous clinical sequelae of hyperphosphatemia include secondary hyperparathyroidism and increased risk of cardiovascular death. Chronic hemodialysis as it is currently practiced in the United States does not remove sufficient phosphate to control serum levels within accepted guidelines. The inadequacy of conventional hemodialysis in removing phosphate mandates the use of phosphate binders in virtually all hemodialysis patients. Despite their proven efficacy, these medications fail to control phosphorous in 70% of hemodialysis patients. Additionally, these medications may have untoward side effects that must be considered since they are typically intended for lifetime use. Quotidian hemodialysis has in previous uncontrolled studies shown promise in reducing serum phosphorus while at the same time reducing or eliminating the need for phosphate binders. Recent results from our group demonstrate for the first time in a controlled fashion the efficacy of short daily dialysis in controlling serum phosphorus. PMID- 15882311 TI - Calcimimetics: a new tool for management of hyperparathyroidism and renal osteodystrophy in patients with chronic kidney disease. AB - Calcimimetics: a new tool for management of hyperparathyroidism and renal osteodystrophy in patients with chronic kidney disease. Epidemiologic, clinical, and basic scientific studies led to an explosion in our understanding of disorders of mineral metabolism in the chronic kidney disease (CKD) patient. These advances are not always translated into improved care of renal osteodystrophy in CKD-5 patients. The introduction of a new class of drugs, calcimimetics, allows improved control of abnormal calcium/phosphorus metabolism. The calcimimetics compliment, rather than replace, current treatment options for secondary hyperparathyroidism in the chronic disease patient. PMID- 15882312 TI - The role of vitamin D in left ventricular hypertrophy and cardiac function. AB - The role of vitamin D in left ventricular hypertrophy and cardiac function. Cardiovascular disease is the leading cause of death among patients with end stage renal disease (ESRD). Traditional cardiac risk factors, as well as other factors specific to the ESRD population such as hyperphosphatemia, elevated calcium and phosphate product, abnormal lipid metabolism, hyperhomocysteinemia, and chronic inflammation play a role in the excessive risk of cardiovascular death in this population. Left ventricular disorders are proven risk factors for cardiac mortality in hemodialysis patients. These disorders are present in incident ESRD patients at rates far above the general population. There is an accumulating body of evidence that suggests that vitamin D plays a role in cardiovascular disease. Abnormal vitamin metabolism, through deficiency of the active form of 1,25-dihydroxyvitamin D(3), and acquired vitamin D resistance through the uremic state, have been shown to be important in ESRD. Vitamin D deficiency has long been known to affect cardiac contractility, vascular tone, cardiac collagen content, and cardiac tissue maturation. Recent studies using vitamin D receptor deficient mice as a model demonstrate a crucial role of vitamin D in regulation of the renin-angiotensin system. Additionally, there is emerging evidence linking treatment with vitamin D to improved survival on hemodialysis and improvement in cardiac function. The emergence of this data is focusing attention on the previously underappreciated nonmineral homeostatic effects of vitamin D that very likely play an important role in the pathogenesis of cardiac disease in ESRD. PMID- 15882313 TI - Dyslipidemia and progression of cardiovascular calcification (CVC) in patients with end-stage renal disease (ESRD). AB - Dyslipidemia and progression of cardiovascular calcification (CVC) in patients with end-stage renal disease (ESRD). Cardiovascular calcification (CVC) is commonly encountered both in the general population as well as in patients with end-stage renal disease (ESRD). The etiology of CVC in patients with ESRD is multifactorial. Despite that, current debate remains narrowly focused on the role of calcium loading from calcium-based phosphate binders (CBPB) in the pathogenesis and progression of CVC. Yet, the alleged link between these binders and CVC has not been substantiated in well-designed controlled trials. In contrast, the purported role of sevelamer, a non-calcium-based phosphate binder, in slowing the progression of CVC in dialysis patients has attracted widespread attention. The beneficial effect of sevelamer on progression of calcification was thought to be due to lower calcium loading during its use. However, an alternative and possibly more likely mechanism involves sevelamer-induced lowering of LDL cholesterol. In this context, previous studies in individuals with normal renal function have documented amelioration of coronary artery calcification (CAC) with reduction of LDL-cholesterol by treatment with HMG-CoA reductase inhibitors (statins). Given that CAC is a well-accepted marker of atherosclerosis, and that high plasma cholesterol concentration is one of the main risk factors for atherosclerosis, then it is not unreasonable to suspect that CAC may be halted or even reversed by lowering of LDL cholesterol level with statin therapy. Unfortunately, the effect of lowering the LDL-cholesterol level on CAC has not been studied in patients with ESRD. Therefore, conclusions about this important topic should await the results of well-designed clinical studies that control for all factors potentially implicated in the CVC burden of patients with ESRD. In this review, I will discuss the role of various potential mechanisms involved in the pathogenesis of CVC in patients with ESRD, and emphasize the role of dyslipidemia and its treatment in this important clinical entity. PMID- 15882314 TI - Evaluation and treatment of coronary artery disease in patients with end-stage renal disease. AB - Evaluation and treatment of coronary artery disease in patients with end-stage renal disease. Patients with end-stage renal disease (ESRD) are at increased risk of death from coronary artery disease (CAD). The metabolic milieu that results from renal dysfunction appears to accelerate the atherosclerotic process by decades in patients with ESRD. The extremely high prevalence of atherosclerosis in patients with ESRD mandates risk factor identification and treatment. Traditionally, CAD in this patient population has been treated conservatively. Analysis of large databases has highlighted the scope and complexity of this problem; nonetheless, there is a paucity of randomized, controlled trials of CAD in patients with ESRD. In this paper the following issues related to evaluation and treatment of patients with chronic kidney disease are addressed: (1) optimal CAD risk management; (2) evaluation for CAD in patients with ESRD, including the identification of coronary calcification; (3) treatment of CAD with medical therapy and revascularization; (4) relative merits of percutaneous coronary intervention versus bypass surgery. In general, an aggressive approach to medical management of CAD is warranted, even in the setting of subclinical CAD. A low threshold for diagnostic testing should be employed in patients with ESRD. When significant CAD is identified, ESRD patients appear to benefit more from revascularization compared to conservative medical management. Thus, if clinically reasonable, patients with ESRD and CAD should be managed aggressively to improve survival and reduce the incidence of future cardiac events. PMID- 15882316 TI - Intensive use of ground water and sustainability. PMID- 15882315 TI - Regulation of PTH synthesis and secretion relevant to the management of secondary hyperparathyroidism in chronic kidney disease. AB - Regulation of PTH synthesis and secretion relevant to the management of secondary hyperparathyroidism in chronic kidney disease. Small decreases in serum Ca(++) and more prolonged increases in serum phosphate (P) stimulate the parathyroid (PT) to secrete parathyroid hormone (PTH), while 1,25(OH)(2)-vitamin D(3) decreases PTH synthesis and secretion. A prolonged decrease in serum Ca(++) and 1,25(OH)(2)D(3), or increase in serum P, such as in patients with chronic renal failure, leads to the appropriate secondary increase in serum PTH. This secondary hyperparathyroidism involves increases in PTH gene expression, synthesis, and secretion, and, if chronic, to proliferation of the parathyroid cells. A low serum Ca(++) leads to an increase in PTH secretion, PTH mRNA stability, and parathyroid cell proliferation. Pi also regulates the parathyroid in a similar manner. The effect of Ca(++) on the parathyroid is mediated by a membrane Ca(2+) receptor (CaR). 1,25(OH)(2)D(3) decreases PTH gene transcription. Ca(2+) and P regulate the PTH gene post-transcriptionally by regulating the binding of parathyroid cytosolic proteins, trans factors, to a defined cis sequence in the PTH mRNA 3'-untranslated region (UTR), thereby determining the stability of the transcript. The parathyroid trans factors and cis elements have been defined. PMID- 15882317 TI - The appropriate use of the Rorabaugh model to estimate ground water recharge. PMID- 15882319 TI - Comment on the "Nothing older than three years". PMID- 15882321 TI - Is current hydrogeologic research addressing long-term predictions? AB - Hydrogeology is a field closely related to the needs of society. Many problems of current national and local interest require predictions of hydrogeological system behavior, and in a number of important cases, the period of prediction is tens to hundreds of thousands of years. It is argued that the demand for such long-term hydrogeological predictions casts a new light on the future needs of hydrogeological research. Key scientific issues are no longer concerned only with simple processes or narrowly focused modeling or testing methods but also with assessment of prediction uncertainties and confidence, couplings among multiple physicochemical processes occurring simultaneously at a site, and the interplay between site characterization and predictive modeling. These considerations also have significant implications for hydrogeological education. With this view, it is asserted that hydrogeological directions and education need to be reexamined and possibly refocused to address specific needs for long-term predictions. PMID- 15882322 TI - Influence of ancient thrust faults on the hydrogeology of the Blue Ridge Province. AB - The Blue Ridge Province contains ubiquitous northeast-southwest-trending thrust faults or smaller thrust "slivers" that greatly impact the nature and character of ground water flow in this region. Detailed investigations at a field site in Floyd County, Virginia, indicate that high-permeability zones occur in the brittle crystalline rocks above these thrust faults. Surface and borehole geophysics, aquifer tests, and chlorofluorocarbon and geochemical data reveal that the shallow saprolite aquifer is separated from the deeper fault-zone aquifer by a low-fracture permeability bedrock confining unit, the hydraulic conductivity of which has been estimated to be six orders of magnitude less than the conductivity of the fault zones at the test site. Within the Blue Ridge Province, these fault zones can occur at depths of 300 m or more, can contain a significant amount of storage, and yield significant quantities of water to wells. Furthermore, it is expected that these faults may compartmentalize the deep aquifer system. Recharge to and discharge from the deep aquifer occurs by slow leakage through the confining unit or through localized breach zones that occur where quartz accumulated in high concentrations during metamorphism and later became extensively fractured during episodes of deformation. The results of this investigation stress the importance of thrust faults in this region and suggest that hydrogeologic models for the Blue Ridge Province include these ancient structural features. Faults in crystalline-rock environments may not only influence the hydrology, they may dominate the flow characteristics of a region. PMID- 15882323 TI - Modeling fracture porosity development using simple growth laws. AB - A model of porosity development has been developed to investigate general relationships between simple fracture aperture growth laws and fracture porosity in evolved fracture arrays in aquifers. The growth of fracture apertures in two dimensional orthogonal arrays with initially spatially uncorrelated lognormal aperture distributions has been studied, where aperture growth rate is proportional to an exponent of the flow rate through each fracture. The evolved arrays show geometrical phase changes as a function of the aperture growth rate exponent, e, and the standard deviation of the initial aperture distribution, sigma(z). Low values of e and sigma(z) lead to bimodal aperture distributions, where apertures parallel to flow are preferentially enlarged. At moderate values of e and sigma(z), there is a transition to a regime of more complex geometries consisting of networks of channel-like structures of preferentially enlarged apertures. At larger values of e, array-spanning channel-like paths of preferentially enlarged apertures develop, where the tortuosity of the channel like paths is a linear function of sigma(z). Following an initial growth phase, during which dynamically stable aperture configurations develop, arrays undergo simple amplification. The geometry of the evolved aperture fields is diverse and they can be highly complex; consequently, parameterization and prediction of their evolution in terms of the initial aperture distributions and growth rate laws is not trivial. PMID- 15882324 TI - Identifying connections in a fractured rock aquifer using ADFTs. AB - Fractured rock aquifers are difficult to characterize because of their extremely heterogeneous nature. Developing an understanding of fracture network hydraulic properties in these aquifers is difficult and time consuming, and field testing techniques for determining the location and connectivity of fractures in these aquifers are limited. In the Clare Valley, South Australia, well interference is an important issue for a major viticultural area that uses a fractured aquifer. Five fracture sets exist in the aquifer, all dipping > 25 degrees . In this setting, we evaluate the ability of steady-state asymmetric dipole-flow tests (ADFTs) to determine the connections between a test well and a set of piezometers. The procedure involves dividing a test well into two chambers using a single packer and pumping fluid from the upper chamber to the lower chamber. By conducting a series of tests at different packer elevations, an "input" signal is generated in fracture zones connected to the test well. By monitoring the "output" response of the hydraulic dipole field at piezometers, the connectivity of the fractures between the test well and piezometers can be determined. Results indicate the test well used in this study is connected in a complex three dimensional geometry, with drawdown occurring above and below areas of potentiometric buildup. The ADFT method demonstrates that the aquifer evaluated in this study cannot be modeled effectively on the well scale using continuum flow models. PMID- 15882325 TI - Evaluation of time-space distributions of submarine ground water discharge. AB - Submarine ground water discharge (SGD) rates were measured continuously by automated seepage meters to evaluate the process of ground water discharge to the ocean in the coastal zone of Suruga Bay, Japan. The ratio of terrestrial fresh SGD to total SGD was estimated to be at most 9% by continuous measurements of electrical conductivity of SGD. Semidiurnal changes of SGD due to tidal effects and an inverse relation between SGD and barometric pressure were observed. Power spectrum density analyses of SGD, sea level, and ground water level show that SGD near shore correlated to ground water level changes and SGD offshore correlated to sea level changes. SGD rates near the mouth of the Abe River are smaller than those elsewhere, possibly showing the effect of the river on SGD. The ratio of terrestrial ground water discharge to the total discharge to the ocean was estimated to be 14.7% using a water balance method. PMID- 15882326 TI - Predicting ground water nitrate concentration from land use. AB - Ground water nitrate concentrations on Nantucket Island, Massachusetts, were analyzed to assess the effects of land use on ground water quality. Exploratory data analysis was applied to historic ground water nitrate concentrations to determine spatial and temporal trends. Maximum likelihood Tobit and logistic regression analyses of explanatory variables that characterize land use within a 1000-foot radius of each well were used to develop predictive equations for nitrate concentration at 69 wells. The results demonstrate that historic nitrate concentrations downgradient from agricultural land are significantly higher than nitrate concentrations elsewhere. Tobit regression results demonstrate that the number of septic tanks and the percentages of forest, undeveloped, and high density residential land within a 1000-foot radius of a well are reliable predictors of nitrate concentration in ground water. Similarly, logistic regression revealed that the percentages of forest, undeveloped, and low-density residential land are good indicators of ground water nitrate concentration > 2 mg/L. The methodology and results outlined here provide a useful tool for land managers in communities with shallow water tables overlain with highly permeable materials to evaluate potential effects of development on ground water quality. PMID- 15882327 TI - 3H/3He age data in assessing the susceptibility of wells to contamination. AB - Regulatory agencies are becoming increasingly interested in using young-ground water dating techniques, such as the 3H/3He method, in assessing the susceptibility of public supply wells (PSWs) to contamination. However, recent studies emphasize that ground water samples of mixed age may be the norm, particularly from long-screened PSWs, and tracer-based "apparent" ages can differ substantially from actual mean ages for mixed-age samples. We present age and contaminant data from PSWs in Salt Lake Valley, Utah, that demonstrate the utility of 3H and 3He measurements in evaluating well susceptibility, despite potential age mixing. Initial 3H concentrations (measured 3H + measured tritiogenic 3He) are compared to those expected based on the apparent 3H/3He age and the local precipitation 3H record. This comparison is used to determine the amount of modern water (recharged after approximately 1950) vs. prebomb water (recharged before approximately 1950) samples might contain. Concentrations of common contaminants were also measured using detection limits generally lower than those used for regulatory purposes. A clear correlation exists between the potential magnitude of the modern water fraction and both the occurrence and concentration of contaminants. For samples containing dominantly modern water based on their initial 3H concentrations, potential discrepancies between apparent 3H/3He ages and mean ages are explored using synthetic samples that are random mixtures of different modern waters. Apparent ages can exceed mean ages by up to 13 years for these samples, with an exponential age distribution resulting in the greatest discrepancies. PMID- 15882328 TI - Calculation of ground water ages--a comparative analysis. AB - Ground water age is a fundamental, yet complex, concept in ground water hydrology. Discrepancies between results obtained through different modeling approaches for ground water age calculation have been reported, in particular, between ground water ages modeled by advection and direct simulation of ground water ages (e.g., age-mass approach), which includes effects of advection and dispersion. Here, through a series of two-dimensional (2D) simulations, the impact of water mixing through advection and dispersion on modeled 14C and directly simulated ground water ages is assessed. Impact of dispersion on modeled ages is systematically stronger in areas where water velocities are smaller and far more pronounced on 14C ages. This effect is also observed in one-dimensional models. 2D simulations show that longitudinal dispersion generally acts as a "source" of 14C, while vertical dispersion acts as a "sink," leading to apparent younger or older modeled 14C ages as compared to advective and directly simulated ground water ages. The presence of permeable and impermeable faults provides an equally important source for discrepancies, leading to major differences in modeled ages among the three methods considered. Overall, our results show that a 14C modeling approach using a solute transport model for calculating ground water age appears to be more reliable in ground water systems without faults and where water velocities are relatively high than in systems that are relatively more heterogeneous and those where faults are present. Among the three modeling approaches considered here, direct simulation of ground water age seems to yield the most consistent results in complex, heterogeneous ground water flow systems, giving a vertical age structure consistent with ages expected from consideration of the flow system. PMID- 15882329 TI - Process-based interpretation of tracer tests in carbonate aquifers. AB - A tracer test in a carbonate aquifer is analyzed using the method of moments and two analytical advection-dispersion models (ADMs) as well as a numerical model. The numerical model is a coupled continuum-pipe flow and transport model that accounts for two different flow components in karstified carbonate aquifers, i.e., rapid and often turbulent conduit flow and Darcian flow in the fissured porous rock. All techniques employed provide reasonable fits to the tracer breakthrough curve (TBC) measured at a spring. The resulting parameter estimates are compared to investigate how each conceptual model of flow and transport processes that forms the basis of the analyses affects the interpretation of the tracer test. Numerical modeling results suggest that the method of moments and the analytical ADMs tend to overestimate the conduit volume because part of the water discharged at the spring is wrongly attributed to the conduit system if flow in the fissured porous rock is ignored. In addition, numerical modeling suggests that mixing of the two flow components accounts for part of the dispersion apparent in the measured TBC, while the remaining part can be attributed to Taylor dispersion. These processes, however, cannot reasonably explain the tail of the TBC. Instead, retention in immobile-fluid regions as included in a nonequilibrium ADM provides a possible explanation. PMID- 15882330 TI - Influence of injection conditions on field tracer experiments. AB - Calibration of ground water transport models is often performed using results of field tracer experiments. However, little attention is usually paid to the influence, on resulting breakthrough curves, of injection conditions and well aquifer interactions, more particularly of the influence of the possible trapping of the tracer in the injection wellbore. Recently, a new mathematical and numerical approach has been developed to model injection conditions and well aquifer interactions in a very accurate way. Using an analytical solution derived from this model, a detailed analysis is made of the evolution of the tracer input function in the aquifer. By varying injection conditions from one simulation to another, synthetic breakthrough curves are generated with the SUFT3D ground water flow and transport finite-element simulator. These tests show clearly that the shape of the breakthrough curves can be dramatically affected by injection conditions. Using generated breakthrough curves as "actual" field results, a calibration of hydrodispersive parameters is performed, neglecting the influence of injection conditions. This shows that neglecting the influence of actual injection conditions can lead to (1) errors on fitted parameters and (2) misleading identification of the active transport processes. Conclusions and guidelines are drawn in terms of proposed methodologies for better controlling the tracer injection in the field, in order to minimize risk of misinterpretation of results. PMID- 15882331 TI - Fractal travel time estimates for dispersive contaminants. AB - Alternative fractional models of contaminant transport lead to a new travel time formula for arbitrary concentration levels. For an evolving contaminant plume in a highly heterogeneous aquifer, the new formula predicts much earlier arrival at low concentrations. Travel times of contaminant fronts and plumes are often obtained from Darcy's law calculations using estimates of average pore velocities. These estimates only provide information about the travel time of the average concentration (or peak, for contaminant pulses). Recently, it has been shown that finding the travel times of arbitrary concentration levels is a straightforward process, and equations were developed for other portions of the breakthrough curve for a nonreactive contaminant. In this paper, we generalize those equations to include alternative fractional models of contaminant transport. PMID- 15882332 TI - On using simple time-of-travel capture zone delineation methods. AB - As part of its Wellhead Protection Program, the U.S. EPA mandates the delineation of "time-of-travel capture zones" as the basis for the definition of wellhead protection zones surrounding drinking water production wells. Depending on circumstances the capture zones may be determined using methods that range from simply drawing a circle around the well to sophisticated ground water flow and transport modeling. The simpler methods are attractive when faced with the delineation of hundreds or thousands of capture zones for small public drinking water supply wells. On the other hand, a circular capture zone may not be adequate in the presence of an ambient ground water flow regime. A dimensionless time-of-travel parameter T is used to determine when calculated fixed-radius capture zones can be used for drinking water production wells. The parameter incorporates aquifer properties, the magnitude of the ambient ground water flow field, and the travel time criterion for the time-of-travel capture zone. In the absence of interfering flow features, three different simple capture zones can be used depending on the value of T . A modified calculated fixed-radius capture zone proves protective when T < 0.1, while a more elongated capture zone must be used when T > 1. For values of T between 0.1 and 1, a circular capture zone can be used that is eccentric with respect to the well. Finally, calculating T allows for a quick assessment of the validity of circular capture zones without redoing the delineation with a computer model. PMID- 15882333 TI - Support vector machines (SVMs) for monitoring network design. AB - In this paper we present a hydrologic application of a new statistical learning methodology called support vector machines (SVMs). SVMs are based on minimization of a bound on the generalized error (risk) model, rather than just the mean square error over a training set. Due to Mercer's conditions on the kernels, the corresponding optimization problems are convex and hence have no local minima. In this paper, SVMs are illustratively used to reproduce the behavior of Monte Carlo based flow and transport models that are in turn used in the design of a ground water contamination detection monitoring system. The traditional approach, which is based on solving transient transport equations for each new configuration of a conductivity field, is too time consuming in practical applications. Thus, there is a need to capture the behavior of the transport phenomenon in random media in a relatively simple manner. The objective of the exercise is to maximize the probability of detecting contaminants that exceed some regulatory standard before they reach a compliance boundary, while minimizing cost (i.e., number of monitoring wells). Application of the method at a generic site showed a rather promising performance, which leads us to believe that SVMs could be successfully employed in other areas of hydrology. The SVM was trained using 510 monitoring configuration samples generated from 200 Monte Carlo flow and transport realizations. The best configurations of well networks selected by the SVM were identical with the ones obtained from the physical model, but the reliabilities provided by the respective networks differ slightly. PMID- 15882334 TI - Contaminant transport models under random sources. AB - While the discussion of model uncertainty has centered on spatial heterogeneity, it is possible that ground water models have not enjoyed much success as predictive tools often because the sources that were eventually imposed in the field differed from those represented in the simulations. This is because deterministic prediction of future conditions is often inaccurate due to the random nature of contaminant sources, in terms of their timing, location, and magnitude. This paper presents a stochastic framework for accommodating random contaminant sources in conventional, deterministic advection-dispersion transport models. The contaminant sources are first classified into two types: those occurring continuously with a deterministic component and random variations and those occurring randomly at instantaneous discrete-time intervals. For the first type, the governing partial differential equation (PDE) is replaced by a stochastic PDE. The random variations are modeled by Gaussian noise or Brownian motion, and the solution is obtained by using Ito's integration technique. For the second type, Markovian analysis is used for discrete-time contamination events. Both approaches use a deterministic transport model to generate response functions at any observation location and time. The response functions are then integrated to yield probabilistic description of contaminant transport, from which key statistical properties such as mean, standard deviation, and confidence interval can be drawn. PMID- 15882335 TI - Comparison of velocity-log data collected using impeller and electromagnetic flowmeters. AB - Previous studies have used flowmeters in environments that are within the expectations of their published ranges. Electromagnetic flowmeters have a published range from 0.1 to 79.0 m/min, and impeller flowmeters have a published range from 1.2 to 61.0 m/min. Velocity-log data collected in five long-screened production wells in the Pleasant Valley area of southern California showed that (1) electromagnetic flowmeter results were comparable within +/-2% to results obtained using an impeller flowmeter for comparable depths; (2) the measured velocities from the electromagnetic flowmeter were up to 36% greater than the published maximum range; and (3) both data sets, collected without the use of centralizers or flow diverters, produced comparable and interpretable results. Although either method is acceptable for measuring wellbore velocities and the distribution of flow, the electromagnetic flowmeter enables collection of data over a now greater range of flows. In addition, changes in fluid temperature and fluid resistivity, collected as part of the electromagnetic flowmeter log, are useful in the identification of flow and hydrogeologic interpretation. PMID- 15882336 TI - Approximate discharge for constant head test with recharging boundary. AB - The calculation of the discharge to a constant drawdown well or tunnel in the presence of an infinite linear constant head boundary in an ideal confined aquifer usually relies on the numerical inversion of a Laplace transform solution. Such a solution is used to interpret constant head tests in wells or to roughly estimate ground water inflow into tunnels. In this paper, a simple approximate solution is proposed. Its maximum relative error is on the order of 2% as compared to the exact analytical solution. The approximation is a weighted mean between the early-time and late-time asymptotes. PMID- 15882337 TI - Longitudinal dispersivity data and implications for scaling behavior. AB - Longitudinal dispersivity (alpha) data were compiled from 109 different authors for different types of geological media. The data were subdivided into different subsets. Dispersivity values for consolidated media were subdivided as basalts, granites, sandstones, and carbonate rocks, while unconsolidated sediments were subdivided into three reliability classes. The data sets provided here may provide ground water practitioners a preliminary guide to estimate dispersivity values at various scales and to guide and verify theories on scaling behavior. Based on the data set presented here, the relationship that empirically best described the dispersivity data in regard to scale of measurement was in the form of a power law. The scaling exponent for consolidated and unconsolidated geological media varied between 0.40 and 0.92, and 0.44 and 0.94, respectively. Higher reliability subsets of data for the unconsolidated sediments and more frequently tested rock formations indicate that the scaling exponent is at the lower end of the observed range, close to 0.5. No significant difference in scaling exponent was found among different media, and no clear evidence exists for the presence of an upper bound or asymptotic behavior on the relationship for any of the analyzed media. PMID- 15882338 TI - The compleat Darcy: new lessons learned from the first English translation of Les Fontaines Publiques de la Ville de Dijon. PMID- 15882340 TI - Immunosenescence: a problem of lymphopoiesis, homeostasis, microenvironment, and signaling. PMID- 15882341 TI - Effects of aging on early B- and T-cell development. AB - Lymphocyte production in the bone marrow and the thymus is reduced during aging, but why this decline occurs has not been fully elucidated. The ability to isolate hematopoietic stem and progenitor cells using sophisticated flow cytometric strategies and to manipulate them in vitro and in vivo has provided insights into the effects of aging on primary lymphopoiesis. These analyses have showed that intrinsic changes in hematopoietic precursors that affect their proliferative potential are one factor that contributes to the age-related decline in B- and T cell production. This and other age-related defects may be exacerbated by changes in the lymphopoietic support potential of the bone marrow and thymic microenvironments as well as by age-induced fluctuations in the production of various endocrine hormones. Particular attention with regard to the latter point has focused on changes in the production of sex steroids, growth hormone, and insulin-like growth factor-I. The present review summarizes recent studies of how age-related perturbations affect primary lymphopoiesis and highlights how the data necessitate the reevaluation of a number of existing paradigms. PMID- 15882342 TI - The aging of early B-cell precursors. AB - B-cell genesis in the bone marrow declines with advancing age. In this review, we discuss our current understanding of why B-cell production rates decline with age with a special emphasis on why age-related factors might target very early lymphoid precursors. We consider the impact of aging on cytokine responsiveness and how current models for lineage relationships for very early B- and T-cell precursors might influence interpretations of experiments addressing age associated declines in B- and T-cell differentiation. This discussion centers on the notion that aging affects events associated with the process by which hematopoietic stem cells are guided toward the B-cell pathway. Finally, we present a model in which the age-associated loss of early B-cell precursors is linked to suboptimal function of key transcriptional regulators of very early B cell development. PMID- 15882343 TI - B cells, E2A, and aging. AB - Both mouse and human exhibit deficiencies in humoral immunity during 'old age'. While alterations in phenotype and function have been well documented, the molecular mechanisms that result in immune senescence remain undefined. B lymphopoiesis is suppressed in senescent mice, which may result from deficits at the pre-B-cell stage or earlier (e.g. pro-B cells). This deficit contrasts with the maintenance of the normal number of total peripheral B lymphocytes in senescent mice. However, mature peripheral B cells in aged mice can exhibit reduced efficiencies of both immunoglobulin isotype switching and somatic hypermutation. The basic helix-loop-helix transcription factor E2A is crucial at several stages of B-lymphocyte differentiation, including the development of pro B and pre-B cells within the bone marrow and in isotype switch and somatic hypermutation among peripheral B cells. Therefore, we have focused on the regulation of E2A expression and function during both B lymphopoiesis and isotype class switching in senescent mice. These studies show that E2A expression is normally under complex control at both post-transcriptional and post translational levels. Alterations in E2A expression at both the B-cell precursor and mature B-cell developmental stages are hypothesized to contribute to defects in humoral immunity during senescence. PMID- 15882344 TI - B cells and aging: gauging the interplay of generative, selective, and homeostatic events. AB - Lymphocyte homeostasis encompasses a continuum of processes that together determine the production, turnover, composition, and representation of lymphocyte pools. These processes include commitment to lymphoid lineages, expansion of progenitor pools, successful transit through intermediate maturation stages, negative and positive selection based on receptor specificity, steady-state maintenance of peripheral lymphocytes, and regulation of antigen-driven activation. Understanding the impact of aging on lymphocyte homeostasis thus requires appreciation of not only the mechanisms responsible for generating and sustaining antigen-reactive B and T cells but also how age-related events can subvert these. Even under the influence of normally operating homeostatic mechanisms, lesions yielding perturbations outside of evolutionarily anticipated boundaries will yield aberrant lymphoid function and representation both upstream and downstream of the primary defect. Accordingly, determining the relative contribution of lineage-intrinsic versus compensatory homoeostatic processes throughout the continuum of lymphoid system development, selection, and maintenance are critical first steps towards understanding age-associated alterations in the immune system. PMID- 15882346 TI - Insights into thymic aging and regeneration. AB - The deterioration of the immune system with progressive aging is believed to contribute to morbidity and mortality in elderly humans due to the increased incidence of infection, autoimmunity, and cancer. Dysregulation of T-cell function is thought to play a critical part in these processes. One of the consequences of an aging immune system is the process termed thymic involution, where the thymus undergoes a progressive reduction in size due to profound changes in its anatomy associated with loss of thymic epithelial cells and a decrease in thymopoiesis. This decline in the output of newly developed T cells results in diminished numbers of circulating naive T cells and impaired cell mediated immunity. A number of theories have been forwarded to explain this 'thymic menopause' including the possible loss of thymic progenitors or epithelial cells, a diminished capacity to rearrange T-cell receptor genes and alterations in the production of growth factors and hormones. Although to date no interventions fully restore thymic function in the aging host, systemic administration of various cytokines and hormones or bone marrow transplantation have resulted in increased thymic activity and T-cell output with age. In this review, we shall examine the current literature on thymic involution and discuss several interventional strategies currently being explored to restore thymic function in elderly subjects. PMID- 15882345 TI - Immunosenescence and macrophage functional plasticity: dysregulation of macrophage function by age-associated microenvironmental changes. AB - The macrophage lineage displays extreme functional and phenotypic heterogeneity, which appears to be because, in large part, of the ability of macrophages to functionally adapt to changes in their tissue microenvironment. This functional plasticity of macrophages plays a critical role in their ability to respond to tissue damage and/or infection and to contribute to clearance of damaged tissue and invading microorganisms, to recruitment of the adaptive immune system, and to resolution of the wound and of the immune response. Evidence has accumulated that environmental influences, such as stromal function and imbalances in hormones and cytokines, contribute significantly to the dysfunction of the adaptive immune system. The innate immune system also appears to be dysfunctional in aged animals and humans. In this review, the hypothesis is presented and discussed that the observed age-associated 'dysfunction' of macrophages is the result of their functional adaptation to the age-associated changes in tissue environments. The resultant loss of orchestration of the manifold functional capabilities of macrophages would undermine the efficacy of both the innate and adaptive immune systems. The macrophages appear to maintain functional plasticity during this dysregulation, making them a prime target of cytokine therapy that could enhance both innate and adaptive immune systems. PMID- 15882347 TI - T cells in aging mice: genetic, developmental, and biochemical analyses. AB - A combination of approaches - gene mapping, biomarker analysis, and studies of signal transduction - has helped to clarify the mechanisms of age-related change in mouse immune status and the implications of immune aging for late-life disease. Mapping studies have documented multiple quantitative trait loci (QTL) that influence the levels of age-sensitive T-cell subsets. Some of these QTL have effects that are demonstrable in young-adult mice (8 months of age) and others demonstrable only in middle-aged mice (18 months). Biomarker studies show that T cell subset levels measured at 8 or 18 months are significant predictors of lifespan for mice dying of lymphoma, fibrosarcoma, mammary adenocarcinoma, or all causes combined. Mice whose immune systems resemble that of young animals, i.e. with low levels of CD4(+) and CD8(+) memory T cells and relatively high levels of CD4(+) T cells, tend to outlive their siblings with the opposite subset pattern. Biochemical analyses show that T cells from aged mice show defects in the activation process within a few minutes of encountering a stimulus and that the defects precede the recognition by the T-cell receptor of agonist peptides on the antigen-presenting cell. Defective assembly of cytoskeletal fibers and hyperglycosylation of T-cell surface glycoproteins contribute to the immunodeficiency state, and indeed treatment with a sialylglycoprotein endopeptidase can restore full function to CD4(+) T cells from aged donors in vitro. PMID- 15882348 TI - In vivo regulation of telomerase activity and telomere length. AB - Telomeres are specialized DNA-protein structures found at the ends of all linear chromosomes. In mammalian cells, they consist of hexanucleotide (TTAGGG) repeats and multiple associated proteins. Telomeres protect the ends of chromosomes and prevent their recognition as DNA breaks. Loss of functional telomere length below a critical threshold can activate programs leading to cell senescence or death. Telomere length represents a balance between the loss of terminal telomeric repeats, which occurs during cell division with incomplete DNA replication, and the addition of telomeric repeats by the unique RNA-dependent DNA polymerase telomerase. Although most somatic cells do not express telomerase, telomerase is induced in lymphocytes at critical stages of development and activation. Telomerase expression thus may prolong the replicative capacity of lymphocytes and thereby enhance their function in immune responses. We have used murine model systems to address two broadly defined questions about lymphocyte telomere biology: how is telomerase physiologically regulated in T cells responding to antigen challenge, and what is the effect of transcriptionally altered telomerase expression on telomere length and, consequently, on immune function? PMID- 15882349 TI - Molecular mechanisms of apoptosis in the cells of the immune system in human aging. AB - Aging is associated with progressive decline in immune functions and increased frequency of infections, autoimmunity, and cancer. Among immune functions, a decline in T-cell functions during aging predominates. In this review, I discuss the molecular signaling of three distinct pathways of apoptosis, namely the death receptor pathway, the mitochondrial pathway, and the most recently described endoplasmic reticulum stress pathway, and the relative sensitivity of naive, central memory, and effector memory CD8(+) T-cell subsets to apoptosis. In addition, I review apoptosis, especially via death receptor pathway, in naive and various memory subsets of CD4(+) and CD8(+) T cells (with primary emphasis on CD8(+) naive and memory subsets) in human aging and discuss the role of apoptosis in immune senescence. PMID- 15882350 TI - Impaired apoptosis and immune senescence - cause or effect? AB - Aged animals and humans exhibit a decreased T-cell activation response although they also exhibit increased susceptibility to responses to self-antigens and a loss of self-tolerance. The age-related alteration in T-cell reactivity, polyclonal expansion of T cells, and enhanced production of autoantibodies may reflect the numerous age-associated alterations in the T-cell arm of the immune system that have been revealed in numerous studies. These studies suggest that subpopulations of T cells are not deleted appropriately in older animals. They further suggest that an age-related impairment of Fas/Fas ligand (FasL)-mediated apoptosis - which plays a major role in activation-induced cell death (AICD) of T cells - may contribute to compromised regulation of the immune system. The likely mechanisms that may lead to impaired induction of FasL in AICD senescent T cells include an age-related shift from the apoptosis-sensitive T-helper 1 cell (Th1) response to the AICD-resistant Th2 response, aberrant T-cell receptor/CD3 downstream-signaling pathways, and altered CD28/B7-mediated T-cell costimulatory signals. Pathologically, accumulation of AICD-senescent T cells is associated with a defective cytotoxic T lymphocyte response and generation of autoreactive T cells. Based on the accumulating evidence, we propose that the emergence of the FasL(lo) AICD-senescent T cells is not only an effect of immune aging but also an important cause of T-cell proliferative senescence in both humans and mice. PMID- 15882351 TI - The role of CD8+ T-cell replicative senescence in human aging. AB - The strict limit in proliferative potential of normal human somatic cells - a process known as replicative senescence - is highly relevant to the immune system, because clonal expansion is fundamental to adaptive immunity. CD8(+) T cells that undergo extensive rounds of antigen-driven proliferation in cell culture invariably reach the end stage of replicative senescence, characterized by irreversible cell-cycle arrest and a critically short telomere length. Cultures of senescent CD8(+) T cells also show resistance to apoptosis, permanent loss of CD28 expression, altered cytokine profiles, reduced ability to respond to stress, and various functional changes. Cells with similar characteristics accumulate during normal aging as well as in younger persons infected with human immunodeficiency virus, suggesting that the process of replicative senescence is not an artifact of cell culture but is also occurring in vivo. Interestingly, in elderly persons, the presence of high proportions of CD8(+) T cells with characteristics of replicative senescence is correlated with reduced antibody responses to vaccines as well as with osteoporotic fractures. CD8(+)CD28(-) T cells also accumulate in patients with certain types of cancer. The emerging picture is that senescent CD8(+) T cells may modulate both immune and non-immune functions, contributing not only to reduced anti-viral immunity but also to diverse age-related pathologies. PMID- 15882352 TI - CD28 extinction in human T cells: altered functions and the program of T-cell senescence. AB - The loss of CD28 expression on T cells is the most consistent biological indicator of aging in the human immune system, and the frequency of CD28(null) T cells is a key predictor of immune incompetence in the elderly. There is also mounting evidence for the high frequency of these unusual T cells among patients with inflammatory syndromes or with chronic infections disproportionate with their age. In these pathological states, CD28(null) T cells likely represent prematurely senescent lymphocytes due to persistent immune activation. Unlike the situation in CD28 gene knockout mice that have anergic CD28(0/0) T cells, human CD28(null) T cells are functionally active, long-lived, oligoclonal lymphocytes that lack or have limited proliferative capacity. Results of replicative senescence studies show that CD28(null) T cells are derived from CD28(+) precursors that have undergone repeated stimulation, indicating that CD28 silencing underlies the program of T-cell aging. Dissection of the machinery regulating CD28 expression is paving the way in elucidating the molecular events leading to immune senescence as well as providing clues into the functional rejuvenation of senescent T cells. PMID- 15882353 TI - Non-malignant clonal expansions of CD8+ memory T cells in aged individuals. AB - CD8(+) T cells provide a major line of defense against intracellular pathogens. Upon encounter with antigen, CD8(+) T cells go through three distinct phases involving proliferation, contraction, and differentiation to become eventually long-lived CD8(+) memory T cells. CD8(+) memory T cells provide long-term protection against infection by intracellular pathogens. CD8(+) memory T-cell proliferation and survival are regulated by many factors, including cytokines, and CD8(+) memory T cells are stably maintained over a period of months to years. In aged humans and mice, however, there are significant alterations to the CD8(+) memory T-cell compartment with frequent development of monoclonal expansions of CD8(+) memory T cells in healthy individuals. Interestingly, CD8(+) clonal expansions are not malignant and do not progress to lymphomas, suggesting that these cells must still be under certain constraints. In this review, we discuss our current understanding of factors that contribute to and regulate these CD8(+) clonal expansions as well as the impact of CD8(+) clonal expansions on immune function of the aged. In addition, we discuss similarities and differences between CD8(+) clonal expansions observed in humans and mice, and we postulate that CD8(+) clonal expansions represent a spectrum of biological outcomes ranging from antigen-driven to antigen-independent phenomena. PMID- 15882354 TI - Gene expression characteristics of CD28null memory phenotype CD8+ T cells and its implication in T-cell aging. AB - Accumulation of CD28(null)CD8(+) T cells is considered as one of the hallmarks of aging in the human immune system. However, the precise changes of CD28(null)CD8(+) T cells, compared to those of the precursor CD28(+)CD8(+) memory T cells, have not been determined. In this study, we present an analysis of the global gene expression profiles of CD28(+) and CD28(null) memory phenotype CD8(+) T cells. These two CD8(+) T subsets exhibited an overall similar gene expression profile with only a few dozen genes that were differentially expressed. A wide range of functions, including co-stimulation, effector activity, signaling, and transcription, were possessed by these differentially expressed genes, reflecting significant functional changes of CD28(null) memory phenotype CD8(+) T cells from their CD28(+) counterparts. In addition, CD28(null) memory CD8(+) T cells expressed several natural killer cell receptors and high levels of granzymes, perforin, and FasL, indicating an increasing capacity for cytotoxicity during memory CD8(+) T-cell aging. Interestingly, in vitro culture of these two subsets with interleukin-15 showed that similar gene expression changes occurred in both subsets. Our analysis provides the gene expression portraits of CD28(null) memory phenotype CD8(+) T cells and alteration from their CD28(+) counterparts and suggests potential mechanisms of T-cell aging. PMID- 15882355 TI - Intrinsic versus environmental influences on T-cell responses in aging. AB - A decline in T-cell responses and a switch to memory T-cell predominance occur with aging. We have used the T-cell receptor (TCR) transgenic mouse model to study age-associated changes in T-cell responses that are a consequence of shifts in subset representation versus changes intrinsic to T cells versus changes in the 'aged' microenvironment. We found that naive transgene-expressing (Tg(+)) CD4(+) T cells from aged mice respond to antigen with reduced interleukin-2 (IL 2) production, decreased cell expansion, and limited differentiation to effectors. Comparable to the characteristic accumulation of memory phenotype T cells in aged humans and conventional rodents, Tg(+) CD4(+) T cells from old OTII and 6.5 TCR transgenic mice acquire a memory phenotype without immunization and become hyporesponsive. The naive Tg(+) CD8(+) T cells from aged 2C mice expressed activation markers, produced IL-2, proliferated, and differentiated into cytotoxic T lymphocytes as efficiently as their young counterparts. Responses by adoptive transferred Tg(+) cells from young mice, immunized in young and old conventional hosts, indicated that the host age influences the onset of cell division, level of cell expansion, and number of cytokine-producing cells. Co transfer of dendritic cells (DCs) from young and less so from aged conventional mice partially restored responses. Furthermore, DCs and T-cell migration to draining lymphoid organs was reduced due to deficiencies intrinsic to aged cells and the aged environment. Thus, alterations in T-cell responses in aging are attributable to intrinsic and environmental influences. PMID- 15882356 TI - The effect of age on the cognate function of CD4+ T cells. AB - With increasing age, the ability to produce protective antibodies in response to immunization declines, resulting in reduced efficacy of vaccination. We have examined how reductions in CD4(+) T-cell function contribute to reduced humoral responses, using a model that allows us to compare identical numbers of antigen specific naive T cells from young and aged T-cell receptor transgenic mice. Naive cells from aged mice exhibit reduced responses, both in vitro and in vivo. In vitro, responses of aged T cells can be enhanced by addition of interleukin (IL) 2. In vivo, using an adoptive transfer model with young hosts, naive cells from aged mice exhibit significant reductions in cognate helper function, leading to reduced B-cell expansion and differentiation. These age-related defects could be overcome by prior in vitro T helper 2 effector generation with aged T cells. This improvement in cognate function of the aged effectors may be related to the enhancement of CD154 expression, which occurs on aged T cells in the presence of exogenous IL-2. We also found no difference in B-cell expansion and differentiation when young cells were transferred to young or aged hosts. Our results indicate that age-related reductions in humoral responses are mainly due to defects in the cognate helper function of naive CD4(+) T cells from aged individuals. PMID- 15882357 TI - The influence of age on immunity to infection with Mycobacterium tuberculosis. AB - Changes within the immune system during aging lead to an elderly population that is both highly susceptible to infectious diseases and unresponsive to typical vaccine protocols. Using the murine model of tuberculosis, we have identified key differences in the generation of T-helper 1 cell immunity between old and young mice, and this information may be important for the design of new vaccines or post exposure therapies to protect the elderly against infectious diseases. In response to infection with Mycobacterium tuberculosis, it has been shown that the generation of antigen-specific CD4(+) T-cell immunity is impaired in old mice. In contrast, recent findings document that old mice display a transient enhanced resistance that occurs within the first 3 weeks of infection. Early resistance was associated with the presence of CD8(+) T cells and their ability to produce interferon-gamma (IFN-gamma) well before their young counterparts. Further investigation into the mechanism by which CD8(+) T cells are induced to secrete IFN-gammain vivo could provide an approach to enhance the effector function of these cells and subsequently protect elderly individuals from respiratory pathogens such as M. tuberculosis. PMID- 15882358 TI - The immune system, amyloid-beta peptide, and Alzheimer's disease. AB - In this review, the case is made that amyloid-beta peptide in the brain of patients with Alzheimer's disease is a primary cause of the disease and that immunotherapy directed against this peptide has the potential to halt and/or reverse disease progression. This supposition is supported by the capacity of anti-beta-amyloid peptide antibodies to prevent or reverse the disease in mouse models of Alzheimer's disease. Furthermore, preliminary results obtained in a small number of patients with Alzheimer's disease are consistent with the observations made in the mouse model of this disease. We review the relationship between the immune system, amyloid-beta peptide, and Alzheimer's disease and the progress made in applying immunotherapy to patients with Alzheimer's disease. PMID- 15882359 TI - Human immunosenescence: is it infectious? AB - Morbidity and mortality due to infectious disease is greater in the elderly than in the young, at least partly because of age-associated decreased immune competence, which renders individuals more susceptible to pathogens. This susceptibility is particularly evident for novel infectious agents such as in severe acute respiratory syndrome but is also all too apparent for common pathogens such as influenza. Many years ago, it was noted that the elderly possessed oligoclonal expansions of T cells, especially of CD8(+) cells. At the same time, it was established that cytomegalovirus (CMV) seropositivity was associated with many of the same phenotypic and functional alterations to T-cell immunity that were being reported as biomarkers associated with aging. It was discovered that CMV was the prime driving force behind most of the oligoclonal expansions and altered phenotypes and functions of CD8 cells. Independently, longitudinal studies of a free-living population of the very old in Sweden over the past decade have led to the emerging concept of an 'immune risk phenotype' (IRP), predicting mortality, which was itself found to be associated with CMV seropositivity. These findings support our hypothesis that the manner in which CMV and the host immune system interact is critical in determining the IRP and hence is predictive of mortality. In this sense, then, we suggest that immunosenescence is contagious. PMID- 15882360 TI - Vitamin E and immune response in the aged: molecular mechanisms and clinical implications. AB - Nutritional status has been indicated as a contributing factor to age-related dysregulation of the immune response. Vitamin E, a lipid-soluble antioxidant vitamin, is important for normal function of the immune cells. The elderly are at a greater risk for vitamin E intake that is lower than recommended levels. Vitamin E supplementation above currently recommended levels has been shown to improve immune functions in the aged including delayed-type hypersensitivity skin response and antibody production in response to vaccination, which was shown to be mediated through increased production of interleukin (IL)-2, leading to enhanced proliferation of T cells, and through reduced production of prostaglandin E(2), a T-cell suppressive factor, as a result of a decreased peroxynitrite formation. Vitamin E increased both cell-dividing and IL-producing capacities of naive T cells, but not memory T cells. The vitamin E-induced enhancement of immune functions in the aged was associated with significant improvement in resistance to influenza infection in aged mice and a reduced risk of acquiring upper respiratory infections in nursing home residents. Further studies are needed to determine the signaling mechanisms involved in the upregulation of naive T-cell function by vitamin E as well as the specific mechanisms involved in reduction of risk for upper respiratory infections. PMID- 15882361 TI - Response of aged mice to primary virus infections. AB - Aging is associated with an increased morbidity to virus infections as well as a delay in clearance of symptoms after infection. Studies of sublethal virus infections of aged mice closely mirror the human situation: there is a delay in clearance of virus. The delay in virus clearance is accompanied by a delay and a decrease in T-cell response, particularly of CD8(+) T cells. Intrinsic alterations of T cells of aged mice contribute to this decrease in virus-specific T-cell response; however, evidence suggests that environmental or innate components of the aged host also influence this age-associated decline in clearance of virus. While the changes in the adaptive immune response have been carefully described, the early events in the generation of the T-cell response after virus infection have received limited attention. Importantly, age associated changes in the innate response to virus infection, particularly production of and response to interferon (IFN)-alpha/beta, cytotoxicity and IFN gamma production by natural killer cells, interleukin-12 induction, and depletion of non-specific T cells early during virus infection need further evaluation. PMID- 15882362 TI - Telenursing and patients' recovery from bypass surgery. AB - AIM: This paper reports the qualitative findings of qualitative interviews conducted as part of a study assessing the effectiveness of structured, postdischarge, telephone intervention for patients and their partners recovering from bypass surgery. The data reported here describe the postoperative recovery experiences of a small sample of patients (n = 10) and the intervention role of the specialist nurse delivering the intervention. BACKGROUND: Reduced length of hospital stay for bypass patients means that there are fewer opportunities to provide necessary information and respond to patient concerns in hospital, and much of the process of wound healing and regaining functioning that would once have taken place in hospital now takes place at home and place additional burdens on patients and their caregivers. METHOD: The study was a randomized controlled trial of a telephone intervention aimed at reducing anxiety for patients experiencing a first bypass, and for their caregivers. The intervention consisted of a series of protocols delivered by a nurse with cardiac experience. It was conducted via telephone at discharge and on days 1, 2, 4, 7 and weeks 2 and 7 postdischarge. In the qualitative component of the study, a purposive sample of telephone calls in the treatment group was analysed and data saturation was achieved with 10 transcripts. FINDINGS: Three major patient concerns emerged: physical, affective and lifestyle changes. Anxiety about speed of discharge was a common concern at discharge and at day 1. Otherwise, physical concerns predominated during the first week. At day 7 and after, a shift began to occur to longer term, future-oriented concerns. By week 7, most patients were planning for return to normal activities, and several were making lifestyle changes. The specialist nurse was able to give personalized health promotion information at a time that was appropriate for individual patients. CONCLUSION: Using a qualitative interview method made it possible to understand patients' concerns and study the working of telenursing interventions in terms of providing timely reassurance and health promotion. Further research is needed to test the generalizability of the findings. PMID- 15882363 TI - Causes and management of patient aggression and violence: staff and patient perspectives. AB - AIM: This paper reports a study of staff and patient perspectives on the causes of patient aggression and the way it is managed. BACKGROUND: The incidence of aggression in healthcare is reportedly on the increase, and concerns about the management of this problem are growing. METHOD: A convenience sample of 80 patients and 82 nurses from three inpatient mental healthcare wards were surveyed using The Management of Aggression and Violence Attitude Scale. A further five patients and five nurses from the same sample participated in a number of follow up interviews. RESULTS: Patients perceived environmental conditions and poor communication to be a significant precursor of aggressive behaviour. Nurses, in comparison, viewed the patients' mental illnesses to be the main reason for aggression, although the negative impact of the inpatient environment was recognized. From interview responses, it was evident that both sets of respondents were dissatisfied with a restrictive and under-resourced provision that leads to interpersonal tensions. CONCLUSION: There are differences between the views of staff and patients about reasons for aggression and its management. Future approaches therefore need to be developed that address these opposing views. For example, training in the use of fundamental therapeutic communication skills was advocated by patients, whilst the need for greater attention to organizational deficits was advocated by nurses. A move away from reliance on the use of medication was also felt to be necessary. Evaluation of local needs and practices must be an integral part of this process. PMID- 15882364 TI - First-time mothers: social support and confidence in infant care. AB - AIM: This paper reports a study whose primary aim was to explore the relationship between social support for first-time mothers and their confidence in infant care practices. A secondary aim was to identify their sources of support in the postnatal period. BACKGROUND: Policy documents emphasize the importance of support for new mothers in the postnatal period in caring for their infants. Nurses/midwives require a working knowledge of how social support influences maternal confidence in infant care practices, specifically during the first 6 weeks postdelivery. METHODS: A descriptive, correlational design was used. A 28 item questionnaire was designed to measure social support in the specific context of first-time motherhood and confidence in infant care practices. Content validity was sought and the instrument demonstrated reliability using Cronbach's alpha. A convenience sample of 135 first-time mothers was recruited and 74% completed questionnaires at 6 weeks after birth. Data were collected in 2000. RESULTS: Appraisal support had a statistically significant moderate relationship with confidence in infant care practices (r = 0.4, P < 0.01). Informational support had a weaker but statistically significant relationship (r = 0.2, P < 0.05). Respondents' primary sources of appraisal support were husbands/partners and their own mothers. Public health nurses and mothers were primary sources of informational support. CONCLUSIONS: First-time mothers' husbands/partners need to become active participants in antenatal and postnatal care. Interdisciplinary educational programmes need to be developed so that public health nurses and midwives work collaboratively in facilitating social support for first-time mothers in caring for their infants. Curricula for public health nurses and midwives need to be evidenced-based with respect to social support. PMID- 15882365 TI - Working with girls living on the streets in East Africa: professionals' experiences. AB - AIM: This paper reports a study elucidating the meaning of caring for girls of the street, as experienced by female staff members working with street children in Eastern Africa. BACKGROUND: The phenomenon of children living on the streets is a global and escalating problem, and girls are presumed to be especially vulnerable. In East Africa, the traditional extended family system is rapidly breaking down and traditional gender values seem to remain. This was the context for investigating female carers' experience of caring for girls. METHOD: Interviews were conducted with 37 project staff members working with children living on the streets in the framework of non-governmental organizations in Kenya, Uganda and Tanzania between 1997 and 1998. Transcribed text from female interviewees (n = 13) working with girls of the street was analysed using a phenomenological-hermeneutic approach. FINDINGS: The meaning of caring for girls of the street for female professional carers in East Africa was comprehensively understood as counselling the girls to integrate the past of their adverse life stories with their present identity. Counselling meant conveying visions for a possible re-direction of the life stories, from being a girl of the street into being an accepted family girl. Caring in this context meant being squeezed between ethical demands and gender values. Experiencing frustration and powerlessness was related to gender structures in society, having to fight the grip of street culture, and a lack of professional tools. Hope and satisfaction were related to success in changing the course of life stories of girls and to seeing possibilities for contributing to empowerment of girls and community members. CONCLUSIONS: Gender issues are critical to care provided to girls of the street. Carers felt that they lacked relevant knowledge and support. Ethical aspects and gender issues in relation to professional care for vulnerable girls ought to be addressed in nursing education and practice, not only for developing countries, but also as a matter of global interest. PMID- 15882366 TI - Patient satisfaction with triage nursing care in Hong Kong. AB - AIM: This paper reports a study to examine the relationship between patient satisfaction and triage nursing care in order to assist nurses in defining more clearly their roles, and ultimately to improve the quality of care delivered to emergency patients. BACKGROUND: Patient satisfaction is considered an important indicator of quality care from the perspective of the consumer and has been widely studied in many settings. However, few studies have examined patient satisfaction with emergency nursing services in the particular area of triage. METHODS: A descriptive, correlational study was conducted in 2001 in one urban acute hospital in Hong Kong using Consumer Emergency Care Satisfaction Scale (CECSS), and patient and nurse demographic data were also collected. Following a power calculation, systematic sampling was carried out, and the final sample consisted of 56 urgent, semi-urgent and non-urgent patients triaged. The response rate was 61%. RESULTS: The majority of the participants were satisfied with their triage nursing care and teaching. However, difficulties were encountered during the data collection process, resulting in a relatively low response rate. Correlational analyses revealed that patient satisfaction with triage nursing care was statistically significantly correlated with age and the type of nursing intervention received. Older people were more satisfied with the teaching offered by triage nurses and patients who had received specific nursing interventions gave more positive ratings on the teaching subscale of the CECSS. There were no statistically significant relationships between patient satisfaction with triage nursing care and nurse characteristics, including gender, work experiences and educational level. CONCLUSIONS: Patients were generally satisfied with the care provided by the triage nurses. Measuring patient satisfaction with triage nursing care remains a major challenge for health care providers in emergency care settings. PMID- 15882367 TI - Long-term sickness absence: women's opinions about health and rehabilitation. AB - AIMS: This paper reports a study to identify associations between ideas of health and rehabilitation in groups of women having first-hand experience of long-term sickness absence. BACKGROUND: As a central part of their work, nurses have to be able to understand people in distress. When someone is unable to work because of illness, nurses are one of the central professional categories involved in their rehabilitation. METHODS: Data were collected by Q-sort grid and biographical interviews from 82 women aged 30-49 years who had either been absent from work because of sickness for 60 days or more, or were receiving a disability pension. The data were first analysed by patterns and structures obtained from a modified factor analysis. The second phase of the analysis dealt with relocating the results from the statistical analyses to their social context. RESULTS: Six opinions representing different conceptions of health, illness, and rehabilitation were identified. These ranged from reflecting high levels of trust in the health care system to reliance on 'nature's course' and emphasizing the meaning of 'feeling all right'. CONCLUSIONS: For women on long-term sick leave, it is reasonable to expect that their communication with rehabilitation professionals will be founded in a mutual understanding of basic concepts. However, the disparate opinions about health and rehabilitation identified in this study show that future studies need to investigate the prospective value of this categorization to see whether and how these conceptions affect rehabilitation practices. PMID- 15882368 TI - Motivational interviewing to promote physical activity for people with chronic heart failure. AB - AIM: This paper reports a study comparing, over a 5-month period, two different methods of increasing physical activity: a traditional exercise programme and one based on motivational interviewing. BACKGROUND: Chronic heart failure is associated with poor quality of life that can be improved by increased physical activity. Patients who are directed to engage in physical activity have a record of low compliance. METHOD: Sixty older aged heart failure patients were randomly assigned to standard care, motivational interviewing or both treatments. The primary outcome was physical activity (kcal/kg/day), with the 6-minute walk test as a secondary outcome. FINDINGS: At entry, no significant differences were observed between the three groups. Following treatment, the 'motivational interviewing' and 'both treatments' groups reported an increase in their level and type of activities, whereas the 'standard care' group did not. All groups significantly increased their 6-minute walk distance. CONCLUSIONS: Motivational interviewing, which incorporates established behaviour change principles and a flexible approach to promotion of activity, increases reported physical activity in older patients with heart failure over a short period. In terms of level and type of activity, this approach gives a better outcome than standard care, and nurses should explore alternative strategies to promote health in this population. PMID- 15882369 TI - Attachment in older adulthood: concept clarification. AB - AIMS: This paper is an analysis to clarify the concept of attachment by: (1) specifying the antecedents of attachment in older adulthood, (2) describing the defining or critical attributes of attachment as relevant for development beyond adulthood, including older adulthood and old age, and (3) specifying the consequences of attachment in older adulthood. BACKGROUND: Attachment is a concept that has been studied both within and outside the discipline of nursing. Most of the scholarly work on the concept has involved investigations of the concept as it applies at very early developmental phases of the lifespan. Despite this concentration of effort on the concept in childhood, theorists have consistently called for attention to the concept in adulthood and older adulthood to advance our understanding of how such close and supportive relationships relate to health. RESULTS: Antecedents of attachment behaviour include any number of fear-provoking or challenging situations, as well as interactions that involve conflict. Illnesses commonly have these characteristics and so are broadly included as antecedents. Defining attributes of attachment behaviour include three distinct behaviours: (1) proximity, or keeping to one or more 'preferred others', (2) protest following involuntary and perceived permanent separation from a preferred other and, (3) the presence of a secure base, viewed as necessary for developmental exploration and growth. Consequences include environmental interaction secure in the belief that a safe haven exists should reliable clues to danger present themselves, and the mobilization of personal and interpersonal assets called developmental resources. CONCLUSIONS: Nursing implications include the ability of nurses to intervene positively with older clients who may be experiencing age-related transitions that affect health. Knowledge about attachment will help nurses mediate the potentially negative impact of health transitions on developmental tasks and promote high-level wellness and successful ageing. PMID- 15882370 TI - Agents of care and agents of the state: bio-power and nursing practice. AB - AIM: This paper presents a conceptual analysis of the concept of bio-power in the context of nursing, including a critique of the widespread rhetoric that nursing is deprived of power and consequently is an apolitical agency. BACKGROUND: Traditionally, power tends to be defined in terms of repression, interdiction and punishment. On the contrary, work by Michel Foucault with regard to bio-power brings into evidence the productive and positive nature of power at the heart of society. Despite being often used by various academic and professional disciplines, the concept of bio-power is rarely cited in nursing. FINDINGS: Nursing as a profession is at the heart of bio-power in that nurses lie at the crossroads between the anatomo-political and bio-political ranges of power over life. They therefore contribute to social regulation through a vast array of diverse political technologies. Nurses are at the flexing point of the state's requirements and of individual and collective aspirations. They occupy a strategic position that allows them to act as instruments of governmentality. Consequently, nurses constitute a fully-fledged political entity making use of disciplinary technologies and responding to state ideologies. CONCLUSION: The concept of bio-power offers a rich theoretical perspective for nursing, as it questions the definition of nursing care as neutral and mainly provided according to patients' best interests. PMID- 15882371 TI - Ecomapping: an innovative research tool for nurses. AB - AIM: This paper explores the use of ecomaps as a research tool for capturing data, using the example of the dynamic nature of social networks from which informal carers of people living with motor neurone disease draw their support. BACKGROUND: The need for social support in relation to health maintenance and disease management has been identified in the literature but little has been published about ways to investigate support networks. Existing discussion in the literature about the use of ecomaps as a clinical tool in social work creates a valuable framework for data collection which can be readily adapted by nurse researchers. METHOD: We used ecomaps as part of a repertoire of research tools to gather data about the social networks of carers of people living with motor neurone disease. Primary carers participated in three interviews and collaborated in ecomap construction over a period of 10 months during 2003. Analytical correlations were made between ecomaps and interview data. FINDINGS: Ecomaps provided a visual means of facilitating discussions around the structure and strength of networks. Being able to represent the social networks visually through ecomapping enabled people to identify each member of the network, examine the strength of each relationship and ascertain the sources of nurture and tension over time. Limitations to this type of data collection arise when participants try to quantify relationships that have been visually produced. CONCLUSION: Ecomapping is a valuable research tool because it provides visual representation of supportive care networks, capturing strategic data through symbols expressing relationships that may be inadequately portrayed in words. The ecomap incorporates the use of consistent symbols that standardize recipient responses, enabling data comparisons to be made. PMID- 15882372 TI - Cognitive behavioural therapy for primary insomnia: a systematic review. AB - AIM: This paper reports a systematic review of seven studies evaluating the efficacy of cognitive behavioural therapy (CBT) for persistent primary insomnia. BACKGROUND: Insomnia is one of the most common health complaints reported in the primary care setting. Although non-pharmacological treatments such as the CBT have been suggested to be useful in combating the persistent insomnia, the efficacy and clinical utility of CBT for primary insomnia have yet to be determined. METHOD: A systematic search of Ovid, MEDLINE, psychINFO, PsycARTICLES, CINAHL, and EMBASE databases of papers published between 1993 and 2004 was conducted, using the following medical subject headings or key words: insomnia, primary insomnia, psychophysiological insomnia, sleep maintenance disorders, sleep initiation disorders, non-pharmacological treatment, and cognitive behavioural therapy. A total of seven papers was included in the review. FINDINGS: Stimulus control, sleep restriction, sleep hygiene education and cognitive restructuring were the main treatment components. Interventions were provided by psychiatrists except for one study, in which the CBT was delivered by nurses. Among beneficial outcomes, improvement of sleep efficacy, sleep onset latency and wake after sleep onset were the most frequently reported. In addition, participants significantly reduced sleep medication use. Some studies gave follow-up data which indicated that the CBT produced durable clinical changes in total sleep time and night-time wakefulness. CONCLUSIONS: These randomized controlled trial studies demonstrated that CBT was superior to any single-component treatment such as stimulus control, relaxation training, educational programmes, or other control conditions. However, heterogeneity in patient assessment, CBT protocols, and outcome indicators made determination of the relative efficacy and clinical utility of the therapy difficult. Therefore, the standard components of CBT need to be clearly defined. In addition, a comprehensive assessment of patients is essential for future studies. PMID- 15882378 TI - Gluten-free diet survey: are Americans with coeliac disease consuming recommended amounts of fibre, iron, calcium and grain foods? AB - OBJECTIVE: This survey was conducted to assess nutrient intakes and food consumption patterns of adults with coeliac disease who adhere to a strict gluten free diet. DESIGN: Three-day estimated self-reported food records were used to assess daily intakes of calories, percent daily calories from carbohydrates, dietary fibre, iron, calcium and grain food servings. SUBJECTS: Volunteers for this survey were recruited through notices placed in coeliac disease support group newsletters, as well as a national magazine for persons with coeliac disease. Forty-seven volunteers met all criteria for participation and returned useable food records. RESULTS: Group mean daily intake of nutrients by gender: Males (n = 8): 2882 calories; 55% carbohydrate; 24.3 g dietary fibre; 14.7 mg iron; 1288.8 mg calcium; 6.6 grain food servings. Females (n = 39): 1900 calories; 52% carbohydrate; 20.2 g dietary fibre; 11.0 mg iron; 884.7 mg calcium; 4.6 grain food servings. Recommended amounts of fibre, iron and calcium were consumed by 46, 44 and 31% of women and 88, 100 and 63% of men, respectively. CONCLUSIONS: Nutrition therapy for coeliac disease has centred around food allowed/not allowed on a gluten-free diet. Emphasis also should be placed on the nutritional quality of the gluten-free diet, particularly as it concerns the iron, calcium and fibre consumption of women. The use of the estimated food record as the dietary survey method may have resulted in the under-reporting of energy intake. Due to the small sample size and possible bias of survey participants, the findings of this survey may not be representative of the larger coeliac community. PMID- 15882379 TI - 'Controlled by food'- lived experiences of coeliac disease. AB - AIM: The aim of this study was to explore the dilemmas experienced by women and men in their everyday lives in relation to their coeliac disease, and to explore the qualities of these dilemmas in relation to specific situations and living conditions. METHODS: We interviewed 43 informants, aged 20-40 years, using 'The Critical Incident Technique'. Interviews focused on situations that gave rise to confusion or discomfort in relation to disease. The interviews were transcribed verbatim and analysed qualitatively. RESULTS: We found 195 dilemmas, experienced in five arenas: food situation at work, during purchases, when travelling, in relation to meals at home and meals outside the home. Emotions, relationships and the management of daily life were the three main categories of dilemmas that emerged through the analysis. Specific emotions were experienced in relation to the disease, such as isolation, shame, fear of becoming contaminated by gluten and worries about being a bother. In the relationship with other persons the informants experienced a number of complicating dilemmas such as unwanted visibility, neglect, being forgotten, disclosure avoidance and risk taking. Dilemmas related to the management of daily life were restricted product choice, double work and constantly being on call. CONCLUSIONS: The lived experiences of coeliac disease were more varied and profound than expected. It is important to take psychological and social aspects into account in the treatment of patients with coeliac disease. PMID- 15882380 TI - The value of empathy in dietetic consultations. A pilot study to investigate its effect on satisfaction, autonomy and agreement. AB - Forty individuals with diabetes and three dietitians completed a questionnaire concerning their consultation. Empathy was examined using the Empathic Communication Coding System (ECCS) (Bylund & Makoul, 2002). The more empathic the professionals' response to emotional opportunities, the more satisfied patients were with their consultations (r = 0.41, d.f. = 15, P = 0.05). There was a nonsignificant trend that the more empathic opportunities that arise during a consultation, the higher the agreement between patient and dietitian on what was discussed (r = 0.28, P = 0.07). The data also suggest that patients reported more autonomy support when they created more empathic opportunities during their consultation (r = -0.29, P = 0.07). This preliminary study suggests that professionals' responses to empathic opportunities may be a useful component of dietetic consultations. PMID- 15882381 TI - Obesity, dietary pattern and physical activity among children in a suburb with a high proportion of immigrants. AB - BACKGROUND: Obesity among children is a growing problem. Interventions should be planned to meet needs in different cultural settings. The objective of this study was to explore dietary patterns, physical activity and perceptions of relationships between life-style and health among children in a suburb with low socio-economic status and a high proportion of immigrants and refugees. METHODS: All children (n = 114) in Grades 5 and 6, aged 11 to 12 years, at a local Swedish school were invited, 112 participated. Food and exercise habits and perceptions on health and body image were assessed by a validated questionnaire and an interview. Height and weight were measured and risk for passing BMI 25 and 30 kg/m(2), respectively at the age of 18 was calculated. RESULTS: Thirty-one percent of the children were obese or overweight. Thirty-four percent spent more than 3 h daily watching TV or using computer, a behaviour, which clustered with excess intake of sweet drinks and habitually skipping breakfast. Only about half of the children believed that their life-style could affect their health. The results show that low socio-economic status and migration are proxies for overweight and obesity. CONCLUSIONS: Obesity, unhealthy living and a low awareness of relations between life-style and health call for action, for culturally sensitive prevention and treatment approaches. PMID- 15882382 TI - Increasing daily fruit and vegetable consumption: what changes do cardiac patients make? AB - BACKGROUND: Interventions targeting fruit and vegetable consumption report significant increases in consumption but do not detail how increases are achieved. This prospective study explored (i) the changes in daily fruit and daily vegetable consumption of cardiac patients participating in an intervention study and (ii) how participants made these changes. METHODS: A total of 120 cardiac patients were asked to increase their daily fruit and vegetable consumption by two portions and to maintain this over 3 months. They were telephoned at 7-, 28- and 90-day follow-up to record daily consumption using a dietary questionnaire; 94 participants completed all parts of the study. RESULTS: Mean reported daily fruit and daily vegetable consumption increased by 1.07 (SD = 1.26) and 0.34 (SD = 0.96) portions, respectively, over 3 months. These increases were statistically significant (P < 0.001) and greatest for participants who reported eating low levels of fruit and vegetables at recruitment. Eating fresh fruit as a snack and at mealtimes were preferred choices for participants. CONCLUSIONS: Providing information and telephone follow-up could be used by busy healthcare professionals instead of face-to-face contact. Interventions to increase total fruit and vegetable consumption could usefully focus on eating fruit. Interventions to increase vegetable consumption need further investigation. PMID- 15882383 TI - The impact of a nutrition education intervention on main meal quality and fruit intake in people with financial problems. AB - BACKGROUND: As part of a course teaching household budgeting, a nutrition education intervention was provided to people with financial problems. The present study aimed at assess the effects of this intervention on the nutritional quality of their main meal and fruit intake, and to collect process information on intervention participation, reactions and opinions. METHODS: For the effect study, a quasi-experimental multiple pre and post-test control group design was used. Telephone dietary recalls were conducted with 35 people in the intervention group and 39 people in the control group. Observations, personal interviews and written questionnaires were used to collect the process data. RESULTS: A significant reduction in saturated fat intake during the mail meal was found. Also, an intervention effect was found for fruit juice consumption, but not for daily fruit intake or vegetable intake during main meal occasions. CONCLUSIONS: The process data revealed some improvements that should be made to the intervention, such as providing more individualized information and tailored recipes. PMID- 15882384 TI - Comparison of energy and protein intakes of older people consuming a texture modified diet with a normal hospital diet. AB - BACKGROUND: There are very few studies looking at the energy and protein requirements of patients requiring texture modified diets. Dysphagia is the main indication for people to be recommended texture-modified diets. Older people post stroke are the key group in the hospital setting who consume this type of diet. The diets can be of several consistencies ranging from pureed to soft textures. OBJECTIVE: To compare the 24-hour dietary intake of older people consuming a texture modified diet in a clinical setting to older people consuming a normal hospital diet. METHOD: Weighed food intakes and food record charts were used to quantify the patients' intakes, which were compared to their individual requirements. RESULTS: The oral intake of 55 patients was measured. Twenty-five of the patients surveyed were eating a normal diet and acted as controls for 30 patients who were prescribed a texture-modified diet. The results showed that the texture-modified group had significantly lower intakes of energy (3877 versus 6115 kJ, P < 0.0001) and protein (40 versus 60 g, P < 0.003) compared to consumption of the normal diet. The energy and protein deficit from estimated requirements was significantly greater in the texture-modified group (2549 versus 357 kJ, P < 0.0001; 6 versus 22 g, P = 0.013; respectively). CONCLUSION: These statistically significant results indicate that older people on texture-modified diets have a lower intake of energy and protein than those consuming a normal hospital diet and it is likely that other nutrients will be inadequate. All patients on texture-modified diets should be assessed by the dietitian for nutritional support. Evidence based strategies for improving overall nutrient intake should be identified. PMID- 15882390 TI - Looking at the future and seeing the past: the challenge of the middle years of parenting a child with intellectual disabilities. AB - BACKGROUND: This paper seeks to understand and conceptualize the experience of mothers of adolescents with intellectual disabilities (IDs) at a time in their lives which others have characterized as 'mid-life' or the 'middle years of parenting'. The concerns of the paper are the lifecourse concerns in mothers' own lives and with biographical elements of becoming and being such a parent. METHODS: Qualitative interviews were conducted with mothers of adolescents with IDs. The average age of mothers was 48 years. Typically parents were interviewed on two to three occasions. RESULTS: The data suggest that despite the difficulties they faced, these parents had constructed a 'life-as-ordinary' in the early phase of their parental careers. They saw themselves as 'ordinary mothers'. However, the social content and events of the middle years of parenting prompt a realization that their lives and, for some, their sense of 'self', are undergoing considerable change. Mothers are forced to look over their lives to find the meaning and significance of these events. For some, there is biographical reinforcement. For others, there is only disruption. DISCUSSION: The overall picture of these years is one of considerable changes and challenges, and underlines the need for a focus on the lifecourse concerns of parents as well as their children. The implications of the data for further research and service development are discussed in the context of identity theory. PMID- 15882391 TI - Parenting stress in mothers of children with an intellectual disability: the effects of parental cognitions in relation to child characteristics and family support. AB - BACKGROUND: Recent theories of stress and coping in parents of children with intellectual disabilities (ID) emphasize the importance of cognitive appraisals in influencing parents' levels of stress and their adaptations to difficulties presented by the children. This study investigated the relationships between parental cognitions, child characteristics, family support and parenting stress. The aspects of cognitions studied were: parenting self-esteem (including efficacy and satisfaction) and parental locus of control. METHODS: The group studied consisted of 46 mothers of children with ID. The Vineland Adaptive Behavior Scales and Maladaptive Behavior Domain were administered by interview. Mothers also completed four questionnaires: the Family Support Scale, the Parenting Sense of Competence Scale, a shortened form of the Parental Locus of Control Scale and the Parenting Stress Index (Short Form). RESULTS: Data were analysed using Pearson's correlation coefficients, partial correlations and a regression analysis. The results indicated that most of the variance in parenting stress was explained by parental locus of control, parenting satisfaction and child behaviour difficulties. Whilst there was also a strong correlation between family support and parenting stress, this was mediated by parental locus of control. CONCLUSIONS: The results demonstrate the potential importance of parental cognitions in influencing parental stress levels. It is argued that these results have implications for clinical interventions for promoting parents' coping strategies in managing children with ID and behavioural difficulties. PMID- 15882392 TI - Do children do what they say? Responses to hypothetical and real-life social problems in children with mild intellectual disabilities and behaviour problems. AB - BACKGROUND: Most research on children's social problem-solving skills is based on responses to hypothetical vignettes. Just how these responses relate to actual behaviour in real-life social situations is, however, unclear, particularly for children with mild intellectual disabilities (MID). METHOD: In the present study, the spontaneous and selected responses of 56 children with MID to hypothetical situations from the Social Problem-Solving Test for children with MID (SPT-MID) were compared to their actual behaviour in comparable staged standardized real life conflict situations. Correlations to externalizing behaviour problems were assessed using the Teacher's Report Form (TRF). RESULTS: The results show children with MID and accompanying externalizing behaviour problems to behave more aggressively in the staged real-life conflicts and provide more spontaneous aggressive responses to the hypothetical vignettes than children with MID and no accompanying externalizing behaviour problems; they did not, however, select more aggressive responses from the hypothetical options provided. A moderate correlation was found between the aggressiveness of the spontaneous responses in the hypothetical situations and actual behaviour in the staged real-life situations. In addition, both the spontaneous aggressive responses under hypothetical circumstances and the actual aggressive behaviour under staged real life circumstances were related to teacher-rated aggressive behaviour in the classroom. CONCLUSIONS: It is concluded that the hypothetical vignettes from the SPT-MID do provide information on both the actual behaviour and knowledge of social problem-solving skills of children with MID. PMID- 15882393 TI - Leisure provision for persons with profound intellectual and multiple disabilities: quality time or killing time? AB - BACKGROUND: Information on the duration, frequency and content of leisure activities for persons with profound intellectual and multiple disabilities (PIMD) in residential facilities during weekends was not available. METHOD: The project was carried out in 2003 and included seven facilities. Interviews were held with direct support professionals of 112 living units. Consequently, the actual leisure provision of 160 persons with PIMD was recorded over a period of four weekends. This study also investigated the relationship between setting characteristics and the distribution of the content, frequency and duration of leisure activities. Age and gender of persons with PIMD were under investigation. RESULTS: A total mean of 3.8 h of leisure activities is provided for during the full weekend, almost half of which includes watching television or listening to music. Leisure activities are almost exclusively offered by professionals. Parents or volunteers only provide a minimum of activities during weekends. The results suggest that the leisure provision for persons with PIMD is severely restricted and not bound to any service provider in particular. Generally, with increasing age the leisure provision for persons with PIMD declines in number and in variety. No effect was found for gender. CONCLUSIONS: Leisure time for persons with PIMD contains more empty hours than quality time. PMID- 15882394 TI - ADHD symptoms and insistence on sameness in Prader-Willi syndrome. AB - BACKGROUND: Apart from a pervasive eating disorder, the Prader-Willi (PWS) syndrome is characterized by a distinct behavioural profile comprising maladaptive behaviours, obsessive-compulsive traits and skin picking, all included in the PWS behavioural phenotype. In this study, we present a further delineation of this characteristic behavioural profile by screening for indices of executive dysfunctions related to attention-deficit/hyperactivity disorder (ADHD), immature compulsive-like adherence to sameness and skin picking, and how these features aggregate into symptom constellations in children and adolescents with PWS. METHOD: Parents of 58 individuals with PWS (aged 5-18 years) participated by completing Childhood Routines Inventory (CRI) and Conners' Parent Rating Scale (CPRS-48). RESULTS: Results showed that indices of ADHD and excessive insistence on sameness were common, comorbid and of early onset. They were both associated with conduct problems. Skin picking, appearing as a single and comorbid symptom, was less associated with childlike compulsions and ADHD related problems. CONCLUSIONS: Findings are discussed in terms of further research in executive dysfunctions in PWS. PMID- 15882395 TI - Risk factors for low bone mineral density in individuals residing in a facility for the people with intellectual disability. AB - BACKGROUND: Individuals with intellectual disability (ID) are known to have a high prevalence of both low bone mineral density (BMD) and fractures with significant attendant morbidity. Effective strategies aimed at reducing fractures will be facilitated by the identification of predisposing risk factors. METHODS: Bone mineral density was measured by quantitative ultrasound of the calcaneus performed on 79 women and 132 men residing in a facility for adults with ID. Multiple variable logistic regression analysis was performed to determine the significance of risk factors for low BMD. RESULTS: Mobility impairment consistently appeared to be a significant risk factor for low BMD regardless of age or sex and especially for middle-aged men with profound ID. Further risk was identified for postmenopausal women taking enzyme inducing anticonvulsant medications and middle-aged men who were either smokers or tended to be short. Hispanic followed by Caucasian origin also put middle-aged males at a greater risk than their African-American counterparts. CONCLUSIONS: Specific risk factors for low BMD, some of which have potential for modification, were identified in the study population. Targeted strategies for risk factor reduction may result in a decrease in the high rate of fractures among these individuals. PMID- 15882396 TI - Endometrial adenocarcinoma presenting in a premenopausal patient with tuberous sclerosis. AB - BACKGROUND: Endometrial adenocarcinoma is very uncommon in women under 40 years of age. CASE: A 39-year-old woman with tuberous sclerosis and severe intellectual disability presented with irregular bleeding unresponsive to oral contraceptive therapy. She was subsequently found to have a deeply invasive endometrial adenocarcinoma. CONCLUSION: Caregivers must pay particular attention to signs and symptoms in non-verbal patients. Persistent irregular bleeding on oral contraceptive therapy warrants additional evaluation. PMID- 15882398 TI - Effects on blood concentrations of certain serum fat-soluble vitamins of long term feeding of dairy cows on a diet supplemented with clinoptilolite. AB - The objective of the experiment was to investigate the effect of clinoptilolite (a natural zeolite) supplementation in the ration of dairy cows on serum beta carotene, vitamins A and E concentrations. Fifty-two clinically healthy Holstein cows were randomly assigned to one of three groups according to their age and parity. The first group (group A, n = 17), was offered a concentrate feed supplemented with 1.25% clinoptilolite. The second group (group B, n = 17), was offered a concentrate feed supplemented with 2.5% clinoptilolite. The third group (group C, n = 18), which served as controls, was offered the same concentrate feed without clinoptilolite supplementation. All cows were fed the above concentrates continuously starting 30 days before the expected parturition up to the end of lactation. Blood samples from individual animals were collected just before the start of experiment, at the day of calving and, thereafter, at monthly intervals. All samples were tested for serum beta-carotene, vitamins A and E concentrations. The results showed that the 1.25 and 2.5% supplementation of clinoptilolite had no adverse effect on serum concentrations of beta-carotene, vitamins A and E. PMID- 15882399 TI - The opioid haemorphin-7 in horses during low-speed and high-speed treadmill exercise to fatigue. AB - The opioid neuropeptide haemorphin-7 was measured, by immunoreactivity, in Standardbred horses during low-speed (7 m/s) and high-speed (10 m/s) endurance exercises, lasting 49-58 and 12-16 min respectively. In parallel, heart rate, muscle temperature and plasma lactate concentrations were measured. The profile of the low-speed exercise showed significantly increased heart rate after 10 min [154 beats per minute (bpm)]. After the exercise, muscle temperature (42.1 degrees C) and plasma lactate (4.8 mmol/l) were significantly increased. The profile of the high-speed exercise was comparatively characterized by a higher increase of heart rate after 5 min (194 bpm) and higher increases of muscle temperature (43.2 degrees C) and lactate levels (15.8 mmol/l) after the exercise. The horses were probably exhausted by glycogen depletion in the low-speed exercise and by muscle pH decrease in the high-speed exercise. Haemorphin-7 increased significantly during the high-speed exercise (274.8 fmol/ml) but not during low speed (108.3 fmol/ml), coincident with the results of lactate. These results suggest that plasma haemorphin-7 is measurable in the horse by immunoreactivity, and that intense exercise stimulates release of this opioid. Such endogenous opioids are most likely involved in regulatory functions associated with pain, physical effort, inflammation, and blood pressure variation in horses, as have been established in other species. PMID- 15882400 TI - Pathogenesis of ascites in broilers raised at low altitude: aetiological considerations based on echocardiographic findings. AB - This study reports novel insight into the aetiology of pulmonary hypertension and ascites in broiler chickens. The scope of measurements was focused on anatomical and functional parameters, and blood flow patterns in leghorns (resistant to ascites), fast-growing broilers (susceptible to ascites), broilers developing ascites, and ascitic broilers evaluated in vivo using echocardiography, and further examined in the context of postmortem findings. Both, in vivo observed features and postmortem findings, showed clear differences between broilers and leghorns, and between normal and ascitic broilers. Abnormalities in the heart chamber geometry and blood flow patterns were detected upon echocardiographic examination in all ascitic broilers. Right and left atrio-ventricular (AV) valve regurgitation were common findings in ascitic broilers and some apparently normal broilers, with left AV valve insufficiency being a predominant feature with respect to degree and frequency of occurrence. Blood flow disturbances were not detected in leghorns. Left ventricular fractional shortening (functional parameter) was considerably reduced (P < 0.01) in ascitic birds (mean: 21.7 +/- 2.0 SE) in comparison with normal broilers (mean: 39.1 +/- 3.6 SE), or leghorns (mean: 43.3 +/- 2.4 SE). The presented findings indicate that pathological and functional changes in the left ventricle and atrium play a significant role in the pathogenesis of ascites in broilers. Severe dilation of the left atrium and pulmonary veins seen on postmortem examination, as well as regurgitant blood flow in the left atrium, demonstrated by Doppler study in ascitic birds, provide evidence that chronically elevated pressure in the left atrium is involved in the aetiology of pulmonary hypertension and ascites in fast-growing broilers. PMID- 15882401 TI - p53 expression in canine lymphoma. AB - The tumour suppressor p53 plays a key role in DNA damage and repair. It is the most frequently altered gene in human cancers and these mutations may implicate the genesis and/or progression of tumours. Mutations of the p53 gene were also found in a number of canine cancers, although it is poorly estimated in canine lymphomas. Thus, the aim of this study was to investigate the p53 status in these types of tumours. We have shown that the expression of p53 in canine lymphomas is rare, however significantly differs between lymphomas of T- and B-cell origin. PMID- 15882402 TI - High prevalence of ulcerative posthitis in Rasa Aragonesa rams associated with a legume-rich diet. AB - This study describes an outbreak of ulcerative posthitis that eventually affected 76 of 80 of rams in a flock of Rasa Aragonesa sheep on a legume-rich diet. Lesions were confined to the prepuce and varied from mild hyperaemia to ulcerations. Corynebacterium renale was isolated from the lesions. Treatment of an initial group of 17 was based on a change of diet and topical treatment with Veterin Banedif with prednisolone once a day for 15 days and was successful. Lesions in this group were completely resolved after 15 days. The relationship between a legume-rich diet, ruminal alkalosis, elevated urine pH and posthitis is discussed. PMID- 15882403 TI - Treatment of segmental tibial defects using acute bone shortening followed by gradual lengthening with circular external fixator. AB - The aim of this study was to clinically and radiographically evaluate acute bone shortening followed by gradual lengthening in the treatment of large segmental tibia defects induced in seven clinically normal dogs. A circular external fixator was assembled with one proximal 5/8-circle ring, one middle ring and one distal ring connected with three rods. Thirty per cent of the tibia and fibula were removed in the middle and distal parts of the diaphyses, between the middle and distal rings. Acute bone shortening with compression of proximal and distal segments was performed. A subperiosteal osteotomy was performed between the half ring and middle ring. Bone distraction started 7 days after surgery; after lengthening, the apparatus was left in place for 14 weeks for consolidation of regenerated bone. The frame was removed at the end of this period, and the dogs observed for four more weeks. Functional results were considered excellent in two, good in three and fair in the other two dogs. Bone regeneration within the distraction gap was obtained 14 weeks after neutral fixation period. We concluded that acute bone shortening followed by gradual lengthening by Ilizarov method can be used to treat extensive tibial defects in dogs, although it presents limb temporary abnormal limb shape and unequal length as early disadvantages. PMID- 15882404 TI - Anti-nociceptive efficacy of carprofen, levomethadone and buprenorphine for pain relief in cats following major orthopaedic surgery. AB - A placebo-controlled, randomized blind study was conducted in cats (n = 60) after fracture repair to compare the analgesic effects as well as the side-effects of carprofen, buprenorphine and levomethadone during a 5-day treatment. Cats with severe shock symptoms or increases in blood urea nitrogen (BUN) and creatinine were excluded from the study. The cats were randomly assigned to four groups (n= 15). In group 1, carprofen was administered upon extubation at an initial dose of 4 mg/kg body weight, followed by one-third of that dose three times daily on days 2 to 5. In group 2, buprenorphine was administered in a single dose of 0.01 mg/kg body weight upon extubation and subsequently every 8 h. Levomethadone (group 3) was applied according to the same scheme at a dosage of 0.3 mg/kg body weight each time. The placebo (group 4) was given at the same time intervals as the opioids. Examinations were carried out prior to anaesthesia, between 30 min and 8 h after extubation, and on the following 4 days, 1 h after administration of the analgesics or the placebo as well as 1 h before the next administration. Pain and sedation evaluation was carried out with a visual analogue system (VAS) and with the aid of a numerical estimation scale (NRS). Pain was also scored by measuring mechanical nociceptive threshold of traumatized tissue. Plasma glucose and cortisol concentration, heart rate, respiration rate, blood pressure and body temperature were measured. Furthermore, a complete blood count and clinical chemistry including BUN, creatinine, alanine aminotransferase (ALT), glutamate dehydrogenase (GLDH), arterial blood pressure (AP), total protein and electrolytes of the cats were checked on the day of admission as well as on the last day of this study (day 5). Defaecation and urination as well as wound healing were monitored. On the basis of the mechanical nociceptive threshold of the traumatized tissue, concentrations of plasma glucose and cortisol and pain assessment using NRS and VAS, carprofen was found to have better anti-nociceptive efficacy when compared with the two opioid analgesics, while the analgesic effect of levomethadone was similar to that of buprenorphine. However, the carprofen group also showed comparably high median NRS and VAS pain scores in addition to occasional broad deviations from the group mean on the first post-operative treatment day. Sedative effects were detected for buprenorphine and levomethadone; in addition, symptoms of central excitation were noted with levomethadone. There was no indication of any clinically relevant respiratory depressive or cardiovascular effects, nor of any undesired renal, gastrointestinal or hepatic effects of the analgesics applied. However, the somewhat insensitive examination methods did not permit sufficient evaluation of side-effects, particularly on the gastrointestinal tract and the kidneys. It was found that carprofen and buprenorphine were well-tolerated analgesics for a 5-day administration in the cat, whereas levomethadone caused central excitation in some cases in the dosage scheme used here. However, it was apparent that none of the tested analgesics induced sufficient analgesia in the post-operative phase. For this reason, suitable methods must be found to improve analgesia, particularly in the immediate post-operative phase. PMID- 15882405 TI - The effect on pregnancy rate of progesterone administration after manual reduction of twin embryos in dairy cattle. AB - Twin pregnancies are undesirable in dairy cattle as they increase the risk of abortion and have many negative effects on the profitability of the herd. The purpose of this study was to evaluate manual reduction of a twin embryo in dairy cows bearing unilateral twins. On day 34 of gestation, 33 cows were assigned to one of three treatment groups (n = 11 for each group): untreated cows (group control), amnion rupture (group AR), and amnion rupture plus treatment (intravaginal progesterone for 28 days) (group ART). A significantly higher (P = 0.0001) pregnancy loss rate was recorded in the AR group (100%, 11/11), than in the ART (54.5%, 6/11) and control (27.3%, 3/11) groups. In the ART group, one embryo survived amnion rupture and the cow bearing it had twins at parturition, while the remaining four cows delivered singletons. Our results suggest that the procedure of rupturing the amnion with progesterone supplementation may provide a satisfactory way for twin reduction in dairy cattle. PMID- 15882406 TI - Effects of low-dose dexamethasone on thymus morphology and immunological parameters in veal calves. AB - Glucocorticoids are often illegally used in association with anabolic steroids as growth promoters in veal calves and beef production. An experimental administration of dexamethasone was carried out in veal calves in order to assess the role of low doses of exogenous glucocorticoids on induction of thymus atrophy and on the immune response. Three groups of five veal calves each were included in this study: group D was administered 0.4 mg/day of dexamethasone-21-phosphate per os for 25 days; group V was administered 2 mg of dexamethasone-21 isonicotinate i.m. at days 14 and 21, and group K served as control. At slaughter, the weight of the thymus was severely reduced in group D and in group V, compared with control animals. Lesions included severe lymphoid depletion and hyperplasia of adipose tissue. In situ evaluation of apoptosis in thymus, showed a reduction of the percentage of positive nuclear areas of animals belonging to group V in comparison with control animals. An overall decrease of lymphocyte proliferative response was detected after treatment with short acting dexamethasone, while antibody response was not affected by treatments. PMID- 15882407 TI - Envelope stress responses and Gram-negative bacterial pathogenesis. AB - The sigma(E), Cpx and Bae envelope stress responses of Escherichia coli are involved in the maintenance, adaptation and protection of the bacterial envelope in response to a variety of stressors. Recent studies indicate that the Cpx and sigma(E) stress responses exist in many Gram-negative bacterial pathogens. The envelope is of particular importance to these organisms because most virulence determinants reside in, or must transit through, this cellular compartment. The Cpx system has been implicated in expression of pili, type IV secretion systems and key virulence regulators, while the sigma(E) pathway has been shown to be critical for protection from oxidative stress and intracellular survival. Homologues of the sigma(E)- and Cpx-regulated protease DegP are essential for full virulence in numerous pathogens, and, like sigma(E), DegP appears to confer resistance to oxidative stress and intracellular survival capacity. Some pathogens contain multiple homologues of the Cpx-regulated, disulphide bond catalyst DsbA protein, which has been demonstrated to play roles in the expression of secreted virulence determinants, type III secretion systems and pili. This review highlights recent studies that indicate roles for the sigma(E), Cpx and Bae envelope stress responses in Gram-negative bacterial pathogenesis. PMID- 15882408 TI - Divided genomes: negotiating the cell cycle in prokaryotes with multiple chromosomes. AB - Historically, the prokaryotic genome was assumed to consist of a single circular replicon. However, as more microbial genome sequencing projects are completed, it is becoming clear that multipartite genomes comprised of more than one chromosome are not unusual among prokaryotes. Chromosomes are distinguished from plasmids by the presence of essential genes as well as characteristic cell cycle-linked replication kinetics; unlike plasmids, chromosomes initiate replication once per cell cycle. The existence of multipartite prokaryotic genomes raises several questions regarding how multiple chromosomes are replicated and segregated during the cell cycle. These divided genomes also introduce questions regarding chromosome evolution and genome stability. In this review, we discuss these and other issues, with particular emphasis on the cholera pathogen Vibrio cholerae. PMID- 15882409 TI - The Bacillus subtilis ydcDE operon encodes an endoribonuclease of the MazF/PemK family and its inhibitor. AB - Operons encoding stable toxins and their labile antidote are widespread in prokaryotes and play important roles in plasmid partitioning and cellular responses to stress. One such family of toxins MazF/ChpAK/PemK encodes an endoribonuclease that inactivates cellular mRNAs by cleaving them at specific, but frequently occurring sites. Here we show that the Bacillus subtilis ydcE gene encodes a member of this family of RNases, which we have called EndoA. Overexpression of EndoA is toxic for bacterial cell growth and this toxicity is reversed by coexpression of the gene immediately upstream, ydcD. Furthermore, YdcD inhibits EndoA activity directly in vitro. EndoA has similar cleavage specificity to MazF and PemK and yields cleavage products with 3'-phosphate and 5'-hydroxyl groups, typical of EDTA-resistant degradative RNases. This is the first example of an antitoxin-toxin system in B. subtilis. PMID- 15882410 TI - Switching aconitase B between catalytic and regulatory modes involves iron dependent dimer formation. AB - In addition to being the major citric acid cycle aconitase in Escherichia coli the aconitase B protein (AcnB) is also a post-transcriptional regulator of gene expression. The AcnB proteins represent a distinct branch of the aconitase superfamily that possess a HEAT-like domain (domain 5). The HEAT domains of other proteins are implicated in protein:protein interactions. Gel filtration analysis has now shown that cell-free extracts contain high-molecular-weight species of AcnB. Furthermore, in vitro and in vivo protein interaction experiments have shown that AcnB forms homodimers. Addition of the iron chelator bipyridyl to cultures inhibited the dimer-dependent readout from an AcnB bacterial two-hybrid system. A similar response was observed with a catalytically inactive AcnB variant, AcnB(C769S), suggesting that the monomer-dimer transition is not mediated by the state of the AcnB iron-sulphur cluster. The iron-responsive interacting unit was accordingly traced to the N-terminal region (domains 4 and 5) of the AcnB protein, and not to domain 3 that houses the iron-sulphur cluster. Thus, it was shown that a polypeptide containing AcnB N-terminal domains 5 and 4 (AcnB5-4) interacts with a second AcnB5-4 to form a homodimer. AcnB has recently been shown to initiate a regulatory cascade controlling flagella biosynthesis in Salmonella enterica by binding to the ftsH transcript and inhibiting the synthesis of the FtsH protease. A plasmid encoding AcnB5-4 complemented the flagella-deficient phenotype of a S. enterica acnB mutant, and the isolated AcnB5 4 polypeptide specifically recognized and bound to the ftsH transcript. Thus, the N-terminal region of AcnB is necessary and sufficient for promoting the formation of AcnB dimers and also for AcnB binding to target mRNA. Furthermore, the relative effects of iron on these processes provide a simple iron-mediated dimerization mechanism for switching the AcnB protein between catalytic and regulatory roles. PMID- 15882411 TI - Copper induction of carotenoid synthesis in the bacterium Myxococcus xanthus. AB - Copper induces a red pigmentation in cells of the bacterium Myxococcus xanthus when they are incubated in the dark, at suboptimal growth conditions. The colouration results from the accumulation of carotenoids, as demonstrated by chemical analysis, and by the lack of a copper effect on M. xanthus mutants affected in known structural genes for carotenoid synthesis. None of several other metals or oxidative agents can mimic the copper effect on carotenoid synthesis. Until now, blue light was the only environmental agent known to induce carotenogenesis in M. xanthus. As happens for the blue light, copper activates the transcription of the structural genes for carotenoid synthesis through the transcriptional activation of the carQRS operon. This encodes the ECF sigma factor CarQ, directly or indirectly responsible for the activation of the structural genes, and the anti-sigma factor CarR, which physically interacts with CarQ to blocks its action in the absence of external stimuli. All but one of the other regulatory elements known to participate in the induction of carotenoid synthesis by blue light are required for the response to copper. The exception is CarF, a protein required for the light-mediated dismantling of the CarR-CarQ complex. In addition to carotenogenesis, copper induces other unknown cellular mechanisms that confer tolerance to the metal. PMID- 15882412 TI - LmxPK4, a mitogen-activated protein kinase kinase homologue of Leishmania mexicana with a potential role in parasite differentiation. AB - Members of the mitogen-activated protein (MAP) kinase cascade are important for the establishment of a Leishmania mexicana infection and are involved in flagellar length control, although the underlying molecular mechanisms remain to be elucidated. This study reports the cloning and characterization of LmxPK4, a MAP kinase kinase homologue of L. mexicana displaying putative plant-like regulatory phosphorylation sites. The recombinant protein has autophosphorylating activity and phosphorylates myelin basic protein. An LmxPK4 gene deletion mutant showed a proliferation defect after infection of macrophages and no or delayed lesion development in mice. Irrespective of the onset of lesion development parasites showed an early and homogeneous lesion development in re-infection experiments. This is indicative for a compensation of the null mutant phenotype. Additionally, this phenotype could be reverted by reintroduction of the wild-type gene into the deletion background. Mutants expressing loss-of-function or N terminally truncated versions of LmxPK4 retained the null mutant phenotype. LmxPK4 is stage-specifically expressed in promastigotes and during differentiation to amastigotes, but is not detectable in amastigotes isolated from the mammalian host. Moreover, its in vitro kinase activity increases with temperature rise up to 40 degrees C. Our results suggest that LmxPK4 is involved in the differentiation process and affects virulence of Leishmania mexicana. PMID- 15882413 TI - Genetic, physiological and biochemical characterization of multiple methanol methyltransferase isozymes in Methanosarcina acetivorans C2A. AB - Biochemical evidence suggests that methanol catabolism in Methanosarcina species requires the concerted effort of methanol:5-hydroxybenzimidazolylcobamide methyltransferase (MtaB), a corrinoid-containing methyl-accepting protein (MtaC) and Co-methyl-5-hydroxybenzimidazolylcobamide:2-mercapto-ethanesulphonic acid methyltransferase (MtaA). Here we show that Methanosarcina acetivorans possesses three operons encoding putative methanol-specific MtaB and corrinoid proteins: mtaCB1, mtaCB2 and mtaCB3. Deletion mutants lacking the three operons, in all possible combinations, were constructed and characterized. Strains deleted for any two of the operons grew on methanol, whereas strains lacking all three did not. Therefore, each operon encodes a bona fide methanol-utilizing MtaB/corrinoid protein pair. Most of the mutants were similar to the wild-type strain, with the exception of the DeltamtaCB1 DeltamtaCB2 double mutant, which grew more slowly and had reduced cell yields on methanol medium. However, all mutants displayed significantly longer lag times when switching from growth on trimethylamine to growth on methanol. This indicates that all three operons are required for wild type growth on methanol and suggests that each operon has a distinct role in the metabolism of this substrate. The combined methanol:CoM methyltransferase activity of strains carrying only mtaCB1 was twofold higher than strains carrying only mtaCB2 and fourfold higher than strains carrying only mtaCB3. Interestingly, the presence of the mtaCB2 and mtaCB3 operons, in addition to the mtaCB1 operon, did not increase the overall methyltransferase activity, suggesting that these strains may be limited by MtaA availability. All deletion mutants were unaffected with respect to growth on trimethylamine and acetate corroborating biochemical evidence indicating that each methanogenic substrate has specific methyltransfer enzymes. PMID- 15882414 TI - The CovR response regulator of group A streptococcus (GAS) acts directly to repress its own promoter. AB - The CovR/S (CsrR/S) two component system is a global regulator of virulence gene expression in the group A streptococcus (GAS, Streptococcus pyogenes). The response regulator, CovR, regulates about 15% of the genes of GAS, including its own operon. Using in vitro DNA binding assays with purified CovR protein, we found that CovR binds a DNA fragment including the covR promoter (Pcov). DNaseI footprint analyses showed that phosphorylation of CovR enhanced and extended the protected regions. The proposed CovR consensus binding sequence (ATTARA) was present at most, but not all protected regions. The effect of replacing the two thymine residues in the consensus binding sequence (CB) with guanine residues was evaluated both in vitro and in vivo. Most, but not all, CB mutations reduced binding of CovR in vitro. Using a transcriptional reporter introduced in single copy into the GAS chromosome, we found that mutations at each CB completely or partially relieved CovR-mediated repression in vivo. This suggests that CovR regulation of Pcov is direct. Further support for this conclusion comes from use of an in vitro GAS transcription system in which CovR was sufficient to mediate repression of Pcov. This repression was enhanced by phosphorylation of the protein. In addition, we found that the CovR binding region overlapping the promoter was essential for wild type repression of Pcov both in vitro and in vivo, suggesting that promoter occlusion is a primary mechanism of Pcov repression by CovR. PMID- 15882415 TI - An altered-specificity DNA-binding mutant of Escherichia coli sigma70 facilitates the analysis of sigma70 function in vivo. AB - The sigma subunit of bacterial RNA polymerase is strictly required for promoter recognition. The primary (housekeeping) sigma factor of Escherichia coli, sigma(70), is responsible for most of the gene expression in exponentially growing cells. The fact that sigma(70) is an essential protein has complicated efforts to genetically dissect the functions of sigma(70). To facilitate the analysis of sigma(70) function in vivo, we isolated an altered-specificity DNA binding mutant of sigma(70), sigma(70) R584A, which preferentially recognizes a mutant promoter that is not efficiently recognized by wild-type sigma(70). Exploiting this sigma(70) mutant as a genetic tool, we establish an in vivo assay for the inhibitory effect of the bacteriophage T4-encoded anti-sigma factor AsiA on sigma(70)-dependent transcription. Our results demonstrate the utility of this altered-specificity system for genetically dissecting sigma(70) and its interactions with transcription regulators. PMID- 15882416 TI - CPCR1, but not its interacting transcription factor AcFKH1, controls fungal arthrospore formation in Acremonium chrysogenum. AB - Fungal morphogenesis and secondary metabolism are frequently associated; however, the molecular determinants connecting both processes remain largely undefined. Here we demonstrate that CPCR1 (cephalosporin C regulator 1 from Acremonium chrysogenum), a member of the winged helix/regulator factor X (RFX) transcription factor family that regulates cephalosporin C biosynthesis, also controls morphological development in the beta-lactam producer A. chrysogenum. The use of a disruption strain, multicopy strains as well as several recombinant control strains revealed that CPCR1 is required for hyphal fragmentation, and thus the formation of arthrospores. In a DeltacpcR1 disruption strain that exhibits only hyphal growth, the wild-type cpcR1 gene was able to restore arthrospore formation; a phenomenon not observed for DeltacpcR1 derivatives or non-related genes. The intracellular expression of cpcR1, and control genes (pcbC, egfp) was determined by in vivo monitoring of fluorescent protein fusions. Further, the role of the forkhead transcription factor AcFKH1, which directly interacts with CPCR1, was studied by generating an Acfkh1 knockout strain. In contrast to CPCR1, AcFKH1 is not directly involved in the fragmentation of hyphae. Instead, the presence of AcFKH1 seems to be necessary for CPCR1 function in A. chrysogenum morphogenesis, as overexpression of a functional cpcR1 gene in a DeltaAcfkh1 background has no effect on arthrospore formation. Moreover, strains lacking Acfkh1 exhibit defects in cell separation, indicating an involvement of the forkhead transcription factor in mycelial growth of A. chrysogenum. Our data offer the potential to control fungal growth in biotechnical processes that require defined morphological stages for optimal production yields. PMID- 15882417 TI - A glutamate-alanine-leucine (EAL) domain protein of Salmonella controls bacterial survival in mice, antioxidant defence and killing of macrophages: role of cyclic diGMP. AB - Signature-tagged transposon mutagenesis of Salmonella with differential recovery from wild-type and immunodeficient mice revealed that the gene here named cdgR[for c-diguanylate (c-diGMP) regulator] is required for the bacterium to resist host phagocyte oxidase in vivo. CdgR consists solely of a glutamate alanine-leucine (EAL) domain, a predicted cyclic diGMP (c-diGMP) phosphodiesterase. Disruption of cdgR decreased bacterial resistance to hydrogen peroxide and accelerated bacterial killing of macrophages. An ultrasensitive assay revealed c-diGMP in wild-type Salmonella with increased levels in the CdgR deficient mutant. Thus, besides its known role in regulating cellulose synthesis and biofilm formation, bacterial c-diGMP also regulates host-pathogen interactions involving antioxidant defence and cytotoxicity. PMID- 15882418 TI - Aspergillus nidulans HOG pathway is activated only by two-component signalling pathway in response to osmotic stress. AB - Genome sequencing analyses revealed that Aspergillus nidulans has orthologous genes to all those of the high-osmolarity glycerol (HOG) response mitogen activated protein kinase (MAPK) pathway of Saccharomyces cerevisiae. A. nidulans mutant strains lacking sskA, sskB, pbsB, or hogA, encoding proteins orthologous to the yeast Ssk1p response regulator, Ssk2p/Ssk22p MAPKKKs, Pbs2p MAPKK and Hog1p MAPK, respectively, showed growth inhibition under high osmolarity, and HogA MAPK in these mutants was not phosphorylated under osmotic or oxidative stress. Thus, activation of the A. nidulans HOG (AnHOG) pathway depends solely on the two-component signalling system, and MAPKK activation mechanisms in the AnHOG pathway differ from those in the yeast HOG pathway, where Pbs2p is activated by two branches, Sln1p and Sho1p. Expression of pbsB complemented the high osmolarity sensitivity of yeast pbs2Delta, and the complementation depended on Ssk2p/Ssk22p, but not on Sho1p. Pbs2p requires its Pro-rich motif for binding to the Src-homology3 (SH3) domain of Sho1p, but PbsB lacks a typical Pro-rich motif. However, a PbsB mutant (PbsB(Pro)) with the yeast Pro-rich motif was activated by the Sho1p branch in yeast. In contrast, HogA in sskADelta expressing PbsB(Pro) was not phosphorylated under osmotic stress, suggesting that A. nidulans ShoA, orthologous to yeast Sho1p, is not involved in osmoresponsive activation of the AnHOG pathway. We also found that besides HogA, PbsB can activate another Hog1p MAPK orthologue, MpkC, in A. nidulans, although mpkC is dispensable in osmoadaptation. In this study, we discuss the differences between the AnHOG and the yeast HOG pathways. PMID- 15882419 TI - Mycobacterium avium 104 deleted of the methyltransferase D gene by allelic replacement lacks serotype-specific glycopeptidolipids and shows attenuated virulence in mice. AB - Mycobacterium avium is a major opportunistic pathogen of AIDS patients in the United States. The understanding of M. avium pathogenesis has been hampered by the inability to create gene knockouts by homologous recombination, an important mechanism for defining and characterizing virulence factors. In this study a functional methyltransferase D (mtfD) gene was deleted by allelic replacement in the M. avium strain 104. Methyltransferase D is involved in the methylation of glycopeptidolipids (GPLs); highly antigenic glycolipids found in copious amounts on the M. avium cell surface. Interestingly, the loss of mtfD resulted in M. avium 104 containing only the non-serotype specific GPL. Results also suggest that the mtfD encodes for a 3-O-methyltransferase. The absence of significant amounts of any serotype-specific GPLs as a consequence of mtfD deletion indicates that the synthesis of the core 3,4-di-O-methyl rhamnose is a prerequisite for synthesis of the serotype-specific GPLs. Macrophages infected with the mtfD mutant show elevated production of tumour necrosis factor-alpha (TNF-alpha) and RANTES compared to control infections. In addition, the M. avium 104 mtfD mutant exhibits decreased ability to survive/proliferate in mouse liver and lung compared to wild-type 104, as assessed by bacterial counts. Importantly, the mtfD mutant complemented with a wild-type mtfD gene maintained an infection level similar to wild-type. These experiments demonstrate that the loss of mtfD results in a M. avium 104 strain, which preferentially activates macrophages in vitro and shows attenuated virulence in mice. Together our data support a role for GPLs in M. avium pathogenesis. PMID- 15882420 TI - A member of the cAMP receptor protein family of transcription regulators in Mycobacterium tuberculosis is required for virulence in mice and controls transcription of the rpfA gene coding for a resuscitation promoting factor. AB - Deletion of gene Rv3676 in Mycobacterium tuberculosis coding for a transcription factor belonging to the cAMP receptor protein (CRP) family caused growth defects in laboratory medium, in bone marrow-derived macrophages and in a mouse model of tuberculosis. Transcript profiling of M. tuberculosis grown in vitro identified 16 genes with significantly altered expression in the mutant compared with the wild type. Analysis of the DNA sequences upstream of the corresponding open reading frames revealed that 12 possessed sequences related to a consensus CRP binding site that could represent the sites of action of Rv3676. These included rpfA, lprQ, whiB1 and ahpC among genes with enhanced expression in the wild type, and Rv3616c-Rv3613c, Rv0188 and lipQ among genes exhibiting enhanced expression in the mutant. The activity of an rpfA::lacZ promoter fusion was lowered in the Rv3676 mutant and by mutation of the predicted Rv3676 binding site. Moreover, the product of Rv3676 (isolated as a TrxA fusion protein) interacted specifically with the rpfA promoter, and binding was inhibited by mutation of the Rv3676 site. Although Rv3676 retains four of the six amino acid residues that bind cAMP in Escherichia coli CRP addition of cAMP did not enhance Rv3676 binding at the rpfA promoter in vitro. In summary, it has been shown that Rv3676 is a direct regulator of rpfA expression, and because rpfA codes for a resuscitation promoting factor this may implicate Rv3676 in reactivation of dormant M. tuberculosis infections. PMID- 15882421 TI - The two-component response regulator PprB modulates quorum-sensing signal production and global gene expression in Pseudomonas aeruginosa. AB - The response regulator PprB and its cognate sensor PprA were recently reported as a two-component regulatory system that controls membrane permeability and antibiotic sensitivity of Pseudomonas aeruginosa. We found that a Tn5 insertion mutation in pprB caused a drastic reduction in virulence factor production and cell motility. A transcriptome analysis revealed that 175 genes were regulated by PprB. Among the 113 PprB-activated genes, 85.5% are known to be activated by N-3 oxo-dodecanoyl-homoserine lactone (OdDHL) and N-butanoyl-homoserine lactone (BHL). In particular, the expression of lasI, rhlI and rhlR, which encode key components of the las and rhl quorum-sensing (QS) systems, were significantly decreased in the pprB mutant. These data suggest that PprB might regulate QS signal production. Measurement of OdDHL and BHL in cultures of the mutant sustained this hypothesis. By using various OdDHL- or BHL-responsive QS reporter systems, including lasB-lacZ, lasI-lacZ and rsaL-lacZ, we found that the mutation in pprB resulted in a large decrease in the sensitivity of P. aeruginosa to exogenous OdDHL. However, there was no difference in sensitivity to BHL. Further analysis showed that the OdDHL influx was significantly reduced in the pprB mutant. We conclude that PprB is a novel QS modulator that positively regulates N acylhomoserine lactone production probably by affecting the OdDHL signal influx and thereby influences global expression of the QS-dependent genes. PMID- 15882422 TI - Reduced expression of antigenic proteins MPB70 and MPB83 in Mycobacterium bovis BCG strains due to a start codon mutation in sigK. AB - Mycobacterium bovis Bacille Calmette-Guerin (BCG) strains are genetically and phenotypically heterogeneous. Expression of the antigenic proteins MPB70 and MPB83 is known to vary considerably across BCG strains; however, the reason for this phenotypic difference has remained unknown. By immunoblot, we separated BCG into high- and low-producing strains. By quantitative reverse transcription polymerase chain reaction (RT-PCR), we determined that transcription of the antigen-encoding genes, mpb70 and mpb83, follows the same strain pattern with mRNA levels reduced over 50-fold in low-producing strains. Transcriptome comparison of the same BCG strains by DNA microarray revealed two gene regions consistently downregulated in low-producing strains compared with high-producing strains, one including mpb70 (Rv2875) and mpb83 (Rv2873) and a second that includes the predicted sigma factor, sigK. DNA sequence analysis revealed a point mutation in the start codon of sigK in all low-producing BCG strains. Complementation of a low-producing strain, BCG Pasteur, with wild-type sigK fully restored MPB70 and MPB83 production. Microarray-based analysis and confirmatory RT-PCR of the complemented strains revealed an upregulation in gene transcription limited to the sigK and the mpb83/mpb70 gene regions. These data demonstrate that a mutation of sigK is responsible for decreased expression of MPB70 and MPB83 in low-producing BCG strains and provide clues into the role of Mycobacterium tuberculosis SigK. PMID- 15882423 TI - Modulating factors for the Pkn4 kinase cascade in regulating 6 phosphofructokinase in Myxococcus xanthus. AB - Myxococcus xanthus, a Gram-negative developmental bacterium, contains a large number of protein Ser/Thr kinases (PSTKs). Among these PSTKs, Pkn4 has been shown to be 6-phosphofructokinase (PFK) kinase. PFK associates with the regulatory domain of Pkn4 (Pkn4RD) and is activated by Pkn4-mediated phosphorylation. The activation of PFK is required to consume glycogen accumulated during early development and is essential for efficient sporulation. Using the yeast two hybrid screen, we identified three new factors, MkapA, MkapB and MkapC, that interact with Pkn4 and each contains well-known protein-protein interaction domains. MkapB contains eight tandem repeats of the TPR (tetratrico peptide repeat) domain and its interaction with Pkn4RD was phosphorylation-dependent. MkapB remained associated with Pkn4RD. As a result, Pkn4 did not interact with PFK and its activation was inhibited. While deletion of the pfk-pkn4 operon did not inhibit fruiting body formation, the spore yield was low. In contrast, a mkapB deletion mutant exhibited a 24 h delay in fruiting body formation, accumulated less glycogen in the stationary phase and gave rise to 3.2% spore formation as opposed to 100% attained with DZF1. In addition to Pkn4, MkapA associated with other membrane-associated PSTKs, Pkn1, Pkn2, Pkn8 and Pkn9, while MkapB associated with Pkn8 and Pkn9, and MkapC with Pkn8. These results indicate that there are complex PSTK networks in M. xanthus that share common modulating factors. PMID- 15882424 TI - Regulation of purine biosynthesis by a eukaryotic-type kinase in Streptococcus agalactiae. AB - Group B streptococci (GBS) are the principal causal agents of human neonatal pneumonia, sepsis and meningitis. We had previously described the existence of a eukaryotic-type serine/threonine kinase (Stk1) and phosphatase (Stp1) in GBS that regulate growth and virulence of the pathogen. Our previous results also demonstrated that these enzymes reversibly phosphorylated an inorganic pyrophosphatase. To understand the role of these eukaryotic-type enzymes on growth of GBS, we assessed the stk1-mutants for auxotrophic requirements. In this report, we describe that in the absence of the kinase (Stk1), GBS are attenuated for de novo purine biosynthesis and are consequently growth arrested. During growth in media lacking purines, the intracellular G nucleotide pools (GTP, GDP and GMP) are significantly reduced in the Stk1-deficient strains, while levels of A nucleotides (ATP, ADP and AMP) are marginally increased when compared with the isogenic wild-type strain. We provide evidence that the reduced pools of G nucleotides result from altered activity of the IMP utilizing enzymes, adenylosuccinate synthetase (PurA) and IMP dehydrogenase (GuaB) in these strains. We also demonstrate that Stk1 and Stp1 reversibly phosphorylate and consequently regulate PurA activity in GBS. Collectively, these data indicate the novel role of eukaryotic-type kinases in regulation of metabolic processes such as purine biosynthesis. PMID- 15882425 TI - Anaerobic regulation by an atypical Arc system in Shewanella oneidensis. AB - Shewanella oneidensis strain MR-1 is well known for its respiratory versatility, yet little is understood about how it regulates genes involved in anaerobic respiration. The Arc two-component system plays an important role in this process in Escherichia coli; therefore, we determined its function in S. oneidensis. arcA from S. oneidensis complements an E. coli arcA mutant, but the Arc regulon in S. oneidensis constitutes a different suite of genes. For example, one of the strongest ArcA-regulated gene clusters in E. coli, sdh, is not regulated by the Arc system in S. oneidensis, and the cyd locus, which is induced by ArcA in E. coli under microaerobic conditions, is repressed by ArcA in S. oneidensis under anaerobic conditions. One locus that we identified as being potentially regulated by ArcA in S. oneidensis contains genes predicted to encode subunits of a dimethyl sulphoxide (DMSO) reductase. We demonstrate that these genes encode a functional DMSO reductase, and that an arcA mutant cannot fully induce their expression and is defective in growing on DMSO under anaerobic conditions. While S. oneidensis lacks a highly conserved full-length ArcB homologue, ArcA is partially activated by a small protein homologous to the histidine phosphotransfer domain of ArcB from E. coli, HptA. This protein alone is unable to compensate for the lack of arcB in E. coli, indicating that another protein is required in addition to HptA to activate ArcA in S. oneidensis. PMID- 15882426 TI - Coupling of multicellular morphogenesis and cellular differentiation by an unusual hybrid histidine protein kinase in Myxococcus xanthus. AB - We describe an unusual hybrid histidine protein kinase, which is important for spatially coupling cell aggregation and sporulation during fruiting body formation in Myxococcus xanthus. A rodK mutant makes abnormal fruiting bodies and spores develop outside the fruiting bodies. RodK is a soluble, cytoplasmic protein, which contains an N-terminal sensor domain, a histidine protein kinase domain and three receiver domains. In vitro phosphorylation assays showed that RodK possesses kinase activity. Kinase activity is essential for RodK function in vivo. RodK is present in vegetative cells and remains present until the late aggregation stage, after which the level decreases in a manner that depends on the intercellular A-signal. Genetic evidence suggests that RodK may regulate multiple temporally separated events during fruiting body formation including stimulation of early developmental gene expression, inhibition of A-signal production and inhibition of the intercellular C-signal transduction pathway. We speculate that RodK undergoes a change in activity during development, which is reflected in changes in phosphotransfer to the receiver domains. PMID- 15882427 TI - Binding of PhoP to promoters of phosphate-regulated genes in Streptomyces coelicolor: identification of PHO boxes. AB - The control of phosphate-regulated genes in Streptomyces coelicolor is mediated by the two-component system PhoR-PhoP. When coupled to the reporter xylE gene the pstS, phoRP and phoU promoters were shown to be very sensitive to phosphate regulation. The transcription start points of the pstS, the phoRP and the phoU promoters were identified by primer extension. phoRP showed a leaderless transcript. The response-regulator (DNA-binding) PhoP protein was overexpressed and purified in Escherichia coli as a GST-PhoP fused protein. The DNA-binding domain (DBD) of PhoP was also obtained in a similar manner. Both PhoP and its truncated DBD domain were found to bind with high affinity to an upstream region of the pstS and phoRP-phoU promoters close to the -35 sequence of each of these promoters. DNase I protection studies revealed a 29 bp protected stretch in the sense strand of the pstS promoter that includes two 11 bp direct repeat units. Footprinting of the bidirectional phoRP-phoU promoter region showed a 51 bp protected sequence that encompasses four direct repeat units, two of them with high similarity to the protected sequences in the pstS promoter. PHO boxes have been identified by alignment of the six direct repeat units found in those promoter regions. Each direct repeat unit adjusts to the consensus G(G/T)TCAYYYR(G/C)G. PMID- 15882428 TI - The pyruvate kinase of Stigmatella aurantiaca is an indole binding protein and essential for development. AB - Myxospore formation of the myxobacterium Stigmatella aurantiaca can be uncoupled from the cooperative development i.e. fruiting body formation, by low concentrations of indole. Two putative indole receptor proteins were isolated by their capacity to bind indole and identified as pyruvate kinase (PK) and aldehyde dehydrogenase. The PK activity of Stigmatella crude extracts was stimulated by indole. Cloning of the PK gene (pykA) and the construction of a pykA disruption mutant strikingly revealed that PK is essential for multicellular development: Fruiting body formation was abolished in the mutant strain and indole-induced spore formation was delayed. The developmental defects could be complemented by insertion of the pykA gene at the mtaB locus of the Stigmatella genome excluding any polar effects of the pykA disruption. PMID- 15882430 TI - CTL-based cancer preventive/therapeutic vaccines for carcinomas: role of tumour associated carbohydrate antigens. AB - In this study, we discuss the use of glycopeptides containing tumour-associated carbohydrate antigens (TACA) as preventive vaccines for carcinomas. The results of our recent studies suggest that CD8(+) cytotoxic T cells are capable of recognizing small TACA in a conventional class I MHC-restricted fashion. TACA specific T-cell receptors are highly degenerate and their fine specificity includes the glycosylated amino acid linker together with the sugar moiety. TF, a disaccharide and Tn, its immediate precursor, are TACA largely expressed in carcinomas that can be successfully used as vaccines when conjugated to designer peptide backbones with optimal binding affinity for class I MHC molecules. PMID- 15882431 TI - The pathophysiology of acute graft-versus-host disease. AB - Despite improvements in allogeneic stem cell transplantation, acute graft-versus host disease (GVHD) remains a significant problem after transplantation, and it is still a major cause of post-transplant mortality. Disease progression is characterized by the differentiation of alloreactive T cells to effector cells leading to tissue damage, recruitment of additional inflammatory cell populations and further cytokine dysregulation. To make the complex process of acute GVHD more explicit, the pathophysiology of acute GVHD is often divided into three different phases. This review summarizes the mechanisms involved in the three phases of acute GVHD. PMID- 15882432 TI - Increased expression of Mycobacterium tuberculosis 19 kDa lipoprotein obliterates the protective efficacy of BCG by polarizing host immune responses to the Th2 subtype. AB - Mycobacterium tuberculosis can not only neutralize immune effector functions, but also has the ability to modulate host-signalling cascades involved in the development of these responses. The 19 kDa antigen (Rv3763), a lipoprotein of M. tuberculosis, elicits high levels of interleukin (IL)-12 from macrophages in addition to its powerful immunomodulatory properties, leading to suppression of antigen-presentation signalling cascades. The present study was aimed at analysing the effect of overexpression of this antigen on the immunostimulatory properties of M. bovis Bacille Calmette-Guerin (BCG). We have constructed a recombinant BCG strain (rBCG19N) producing higher levels of the 19 kDa antigen in both the cytoplasmic (approximately eightfold) and extracellular (approximately fivefold) fractions as compared to the wildtype BCG. Immunization of mice with rBCG19N elicited high levels of interferon-gamma (IFN-gamma) and relatively low levels of IL-10 against the purified 19 kDa antigen. However, in response to total BCG sonicate, mice immunized with rBCG19N produced significantly high levels of IL-10 with relatively very low levels of IFN-gamma. This polarization of the host immune responses towards T-helper 2 subtype resulted in complete abrogation of the protective efficacy of BCG, when rBCG19N was used as a live vaccine against M. tuberculosis challenge in guinea pigs. PMID- 15882433 TI - Induction of oral tolerance in experimental Sjogren's syndrome autoimmunity. AB - Previous studies have showed that immunization with peptides from Ro 60 results in Sjogren's syndrome (SS)-like condition in BALB/c mice. We hypothesized that oral feeding with Ro 60 peptide or Ro 60 would prevent the disease. Four groups (each consisting of 10) of BALB/c mice were used. Group I-III were immunized with Ro 274 peptide. Group IV mice were administered adjuvant only. Group II mice were fed orally with Ro 274 peptide and Group III with Ro 60 for 5 days before immunization. There was a significant reduction in the binding of sera from both Group II and Group III mice to most of the Ro multiple antigenic peptides bound by Group I mice. In Group III mice, salivary flow was maintained above that of the Group I mice (average: 117.5 versus 58.6 microl; t = 2.7; P = 0.02). Salivary infiltrates were drastically decreased in the Ro peptide or Ro 60-fed groups, compared to non-tolerized group. Two of eight mice in Group II and 3/6 mice in Group III had no infiltrates, whereas all eight mice studied in Group I had a significant number of infiltrates. Thus, epitope spreading was prevented, lymphocytic infiltration was blocked and saliva flow was restored by means of oral feeding of either Ro 274 or Ro 60 in this animal model of SS. PMID- 15882434 TI - Mannan-binding lectin recognizes structures on ischaemic reperfused mouse kidneys and is implicated in tissue injury. AB - Organ damage as a consequence of ischaemia and reperfusion (I/R) is a major clinical problem in an acute renal failure and transplantation. Ligands on surfaces of endothelial cells that are exposed due to the ischaemia may be recognized by pattern recognition molecules such as mannan-binding lectin (MBL), inducing complement activation. We examined the contribution of the MBL complement pathway in a bilateral renal I/R model (45 min of ischaemia followed by 24 h of reperfusion), using transgenic mice deficient in MBL-A and MBL-C [MBL double knockout (MBL DKO)] and in wildtype (WT) mice. Kidney damages, which were evaluated by levels of blood urea nitrogen (BUN) and creatinine, showed that MBL DKO mice were significantly protected compared with WT mice. MBL DKO mice, reconstituted with recombinant human MBL, showed a dose-dependent severity of kidney injury increasing to a comparable level to WT mice. Acute tubular necrosis was evident in WT mice but not in MBL DKO mice after I/R, confirming renal damages in WT mice. MBL ligands in kidneys were observed to be present after I/R but not in sham-operated mice. C3a (desArg) levels in MBL DKO mice were decreased after I/R compared with that in WT mice, indicating less complement activation that was correlated with less C3 deposition in the kidneys of MBL DKO mice. Our data implicate a role of MBL in I/R-induced kidney injury. PMID- 15882435 TI - Treatment of experimental arthritis with M2000, a novel designed non-steroidal anti-inflammatory drug. AB - The current study was planned to explore the therapeutic potency of M2000 (beta-D mannuronic acid), a novel designed non-steroidal anti-inflammatory drug (NSAID) in adjuvant-induced arthritis model. Arthritis was induced in Lewis rats by a single intradermal injection (0.1 ml) of heat-killed Mycobacterium tuberculosis (0.3 mg) in Freund's incomplete adjuvant into the right footpad. Fourteen days after injection of adjuvant, the contralateral left footpad volume was measured. The animals with paw volumes 0.37 ml greater than normal paws were then randomized into treatment groups. Orally and intraperitoneally administrations of test drugs (M2000, 40/mg/kg/day and indomethacin, 2/mg/kg/day) were started on day 15 post-adjuvant injection and continued until final assessment on day 25. The left hind limb was removed for histological evaluation. The WEHI-164 cell line was used for assaying tolerability and matrix metalloproteinase type 2 (MMP 2) activity. MMP-2 activity was assessed using zymography. Pharmacotoxicology study was carried out on animal models based on the evaluation of serum and urine determinants, histology of kidney, gastrointestinal tolerability and body temperature. Results showed that the orally administration as well as intraperitoneally injection of M2000 to arthritic rats induced a significant reduction in paw oedema. Histopathological assessment showed a reduced inflammatory cells infiltrate in joints of treated rats, as well as the number of osteoclasts present in the subchondral bone, tissue oedema and bone erosion in the paws were markedly reduced following M2000 therapy. Cytotoxicity analysis of M2000 showed a much higher tolerability compared with other tested drugs (diclofenac, piroxicam and dexamethasone). The inhibitory effect of M2000 in MMP 2 activity was significantly greater than that of dexamethasone and of piroxicam at a concentration of 200 microg/ml. Moreover, the toxicological study revealed that M2000 had no influence on serum (blood urea nitrogen, creatinine, triglyceride and cholesterol) and urine (urea and urinary protein excretion) determinants, glomerular histology and body temperature in normothermic rats and had no ulcerogenic effects on rats' stomach. Our data show that M2000, as a novel NSAID, could be strongly suggested as the safest anti-inflammatory drug for long term administration. PMID- 15882436 TI - Inhibitory effects of anti-oxidants on apoptosis of a human polyclonal T-cell line, MT-2, induced by an asbestos, chrysotile-A. AB - To clarify the effects of silica and silicates on cellular features of lymphocytes, a human T-lymphotropic virus type-1-immortalized polyclonal T-cell line, MT-2, was exposed to various concentrations of chrysotile-A, an asbestos classified as silicate. MT-2 cells underwent apoptosis in a dose- and time dependent manner. The mitochondrial apoptotic pathway was activated during chrysotile-A-induced apoptosis of MT-2 cells, because of the phosphorylation of JNK and p38, increase of BAX and release of cytochrome-c from mitochondria to cytoplasma. In addition, anti-oxidants such as hydroxyl-radical excluders and capturers of superoxide and inhibitors of superoxide production effectively reduced the size of the apoptotic fraction in MT-2 cells cultured with chrysotile A. These results indicate that the activation of reactive oxygen species may play a central role in asbestos-induced T-cell apoptosis, and anti-oxidants may help to prevent complications of pneumoconiosis. PMID- 15882437 TI - Quantification of tissue inhibitor of metalloproteinases 2 in plasma from healthy donors and cancer patients. AB - Tissue inhibitor of metalloproteinases (TIMP)-2 is a highly conserved molecule, which binds both active and latent matrix metalloproteinase (MMP)-2. TIMP-2 is also involved in the activation of MMP-2 on the cell surface. A quantitative enzyme-linked immunosorbent assay (ELISA) was established and optimized for measurement of TIMP-2 in plasma. The capturing antibody in the ELISA was a monoclonal, while the detecting antibody was a chicken polyclonal antibody recognizing the native form of human TIMP-2. The levels of TIMP-2 were measured in ethylenediaminetetraacetic acid (EDTA) and citrate plasma from healthy donors. The median values were determined as 163 ng/ml (n = 186) with a range of 109-253 ng/ml for EDTA plasma and 139 ng/ml (n = 77) with a range of 95-223 ng/ml for citrate plasma. The TIMP-2 concentration in citrate plasma from 15 patients with advanced, stage IV breast cancer had a median value of 160 ng/ml, only slightly higher but statistically distinguishable from the level found in citrate plasma from the healthy donors. In addition, the TIMP-2 concentration in EDTA plasma from colorectal cancer patients revealed a significantly higher level in plasma from patients with Dukes stage A (P = 0.01) compared with patients with more advanced Dukes stages. PMID- 15882438 TI - Circulating epstein-barr virus in children living in malaria-endemic areas. AB - Children living in malaria-endemic regions have high incidence of Burkitt's lymphoma (BL), the aetiology of which involves Plasmodium falciparum malaria and Epstein-Barr virus (EBV) infections. Acute malarial infection impairs the EBV specific immune responses with the consequent increase in the number of EBV carrying B cells in the circulation. To further understand the potential influence of malarial infection on the EBV persistence in children living in malaria-endemic areas, we studied the occurrence and quantified cell-free EBV-DNA in plasma from 73 Ghanaian children with and without acute malarial infection. Viral DNA was detected in 40% of the samples (47% in the malaria-infected and 34% in the nonmalaria group) but was absent in plasma from Ghanaian adults and healthy Italian children. These findings provide evidence that viral reactivation is common among children living in malaria-endemic areas, and may contribute to the increased risk for endemic BL. The data also suggest that the epidemiology of EBV infection and persistence varies in different areas of the world. PMID- 15882439 TI - Childhood levels of immunoglobulins and mannan-binding lectin in relation to infections and allergy. AB - Respiratory tract infections, allergies and otitis media are common problems in early childhood. Our aim was to evaluate in a longitudinal community-based cohort study the association between maturation of immunoglobulin (Ig) and mannan binding lectin (MBL) responses and disease manifestations in the first 4 years of life. Sustained low levels of IgA proved the strongest single indicator of susceptibility for recurrent otitis media (P = 0.008) and respiratory tract infections (P = 0.02), and this condition was also associated with low production of IgG subclasses. About 7% of the cohort had sustained low levels of MBL (<0.4 mg/l). Low MBL did not predispose to any ailments studied, but children with low IgA and recurrent otitis media had relatively low MBL at birth, which failed to increase during the study period and was significantly reduced at the age of 4 years (P = 0.04). MBL levels increased from birth to 2 years (P < 0.0001) and were higher in children than in adults (P = 0.001). The increase was 1.9-fold in children with no recorded clinical events and 1.7-fold in children with asthma or infections, but significantly lower, 1.2-fold, in children with recurrent otitis media. Low levels of IgA within the normal range may reveal disease susceptibility not detected by conventional criteria. Slow maturation of Ig appears to be the main factor of susceptibility during childhood, but a strong corollary role for MBL is indicated by the high levels produced during childhood as well as the precipitation of disease in children with low levels of MBL and Ig. PMID- 15882441 TI - E3 ubiquitin ligases as regulators of membrane protein trafficking and degradation. AB - Ubiquitination is a regulated post-translational modification that conjugates ubiquitin (Ub) to lysine residues of target proteins and determines their intracellular fate. The canonical role of ubiquitination is to mediate degradation by the proteasome of short-lived cytoplasmic proteins that carry a single, polymeric chain of Ub on a specific lysine residue. However, protein modification by Ub has much broader and diverse functions involved in a myriad of cellular processes. Monoubiquitination, at one or multiple lysine residues of transmembrane proteins, influences their stability, protein-protein recognition, activity and intracellular localization. In these processes, Ub functions as an internalization signal that sends the modified substrate to the endocytic/sorting compartments, followed by recycling to the plasma membrane or degradation in the lysosome. E3 ligases play a pivotal role in ubiquitination, because they recognize the acceptor protein and hence dictate the high specificity of the reaction. The multitude of E3s present in nature suggests their nonredundant mode of action and the need for their controlled regulation. Here we give a short account of E3 ligases that specifically modify and regulate membrane proteins. We emphasize the intricate network of interacting proteins that contribute to the substrate-E3 recognition and determine the substrate's cellular fate. PMID- 15882442 TI - A novel hook-related protein family and the characterization of hook-related protein 1. AB - The spatial organization of organelles within a cell is dependent on microtubules. Recently, members of the Hook family of proteins have been proposed to function in linking organelles to microtubules. We report the identification of a completely novel protein family, the Hook-related protein (HkRP) family, from which the Hook proteins have diverged. Bioinformatic analysis of the HkRP family revealed several conserved domains, including a unique C-terminal HkRP domain. The central region of each protein is comprised of an extensive coiled coil domain, and the N-terminus contains a putative microtubule-binding domain. This domain has been shown to bind microtubules in the Hook protein and show that the HkRP1 protein is microtubule-associated. While endogenous HkRP1 has no distinct organelle association, expression of the C-terminal membrane-binding domain suggests a function of the HkRP1 in early endosome. Ultrastructural studies reveal that expression of the C-terminal HkRP1 domain causes an accumulation of internal membranes with an electron-dense coat. Co-localization studies show a concomitant redistribution of the early endosome marker sorting nexin 1 but not the early endosome antigen-1 (EEA1). The steady-state distribution of the epidermal growth factor receptor is also specifically disrupted by expression of the C-terminal domain. We propose that HkRP1 is involved in the process of tubulation of sorting nexin-1 positive membranes from early endosome subdomains. PMID- 15882443 TI - The Listeria protein internalin B mimics hepatocyte growth factor-induced receptor trafficking. AB - Increased hepatocyte growth factor receptor (HGFR) signaling correlates closely with neoplastic invasion and metastatic potential of many human cancers. Hepatocyte growth factor receptor signaling is initiated by binding the physiological ligand HGF or the internalin B (InlB) protein of Listeria monocytogenes. Subsequent degradation of endocytosed HGFR terminates receptor signaling. Previously reported discrepancies in InlB and HGF-induced HGFR signaling could reflect differences in receptor internalization and degradation in response to these distinct ligands. We report that soluble InlB and HGF are mechanistically equivalent in triggering clathrin-dependent endocytosis and lysosomal degradation of HGFR. After internalization, InlB and HGF colocalize with Rab5, EEA1 and the transferrin receptor in classical early endosomes. Hepatocyte growth factor receptor internalization was prevented by overexpression of dominant negative mutants of dynamin 1 and epidermal growth factor phosphorylation substrate 15, but not caveolin 1, the GTPase Arf6 or the cholesterol-chelating drug Nystatin. Thus, HGFR internalization is principally clathrin-mediated and is not regulated by clathrin- independent pathways. Phosphatidylinositol 3-kinase signaling and HGF-regulated tyrosine kinase substrate were not required for ligand-triggered internalization of HGFR but were essential for subsequent lysosomal degradation. Thus, soluble InlB and HGF induce HGFR endocytosis and degradation by indistinguishable mechanisms, suggesting that InlB may be exploited to regulate pathogenic HGFR signaling. PMID- 15882444 TI - An extended tyrosine-targeting motif for endocytosis and recycling of the dense core vesicle membrane protein phogrin. AB - Integral membrane proteins of neuroendocine dense-core vesicles (DCV) appear to undergo multiple rounds of exocytosis; however, their trafficking and site of incorporation into nascent DCVs is unclear. Previous studies with phogrin (IA 2beta) identified sorting signals in the luminal domain that is cleaved post translationally; we now describe an independent DCV targeting motif in the cytosolic domain that may function at the level of endocytosis and recycling. Pulse-chase radiolabeling and cell surface biotinylation experiments in the pituitary corticotroph cell line AtT20 showed that the mature 60/65 kDa form that resides in the DCV is generated by limited proteolysis in a post-trans Golgi network compartment with similar kinetics to the formation of the principal cargo, ACTH. Phogrin is exposed on the cell surface in response to stimuli and progressively internalized to a perinuclear compartment that overlaps with recycling endosomes marked by transferrin. Chimeric molecules of phogrin transmembrane and cytosolic sequences with the interleukin-2 receptor alpha chain (Tac) were sorted to DCVs through the action of an extended tyrosine-based motif Y(654)QELCRQRMA located in a 27aa sequence adjacent to the membrane-spanning domain. A 36aa domain terminating in this sequence conferred DCV localization to Tac in the absence of any other cytosolic or luminal phogrin components. The endocytosis and DCV targeting of phogrin Y(654) > A mutants correlated with the impaired binding of the phogrin cytosolic tail to the micro-subunit of the AP2 adaptor complex in vitro. PMID- 15882445 TI - HIV-1 trafficking to the dendritic cell-T-cell infectious synapse uses a pathway of tetraspanin sorting to the immunological synapse. AB - Dendritic cells (DCs) are essential components of the early events of HIV infection. Here, we characterized the trafficking pathways that HIV-1 follows during its capture by DCs and its subsequent presentation to CD4(+) T cells via an infectious synapse. Immunofluorescence microscopy indicates that the virus containing compartment in mature DCs (mDCs) co-labels for the tetraspanins CD81, CD82, and CD9 but contains little CD63 or LAMP-1. Using ratio imaging of pH reporting fluorescent virions in live DCs, we show that HIV-1 is internalized in an intracellular endocytic compartment with a pH of 6.2. Significantly, we demonstrate that the infectivity of cell-free virus is more stable at mildly acidic pH than at neutral pH. Using electron microscopy, we confirm that HIV-1 accumulates in intracellular vacuoles that contain CD81 positive internal membranes but overlaps only partially with CD63. When allowed to contact T cells, HIV-1-loaded DCs redistribute CD81, and CD9, as well as internalized HIV-1, but not the immunological synapse markers MHC-II and T-cell receptor to the infectious synapse. Together, our results indicate that HIV-1 is internalized into a non-conventional, non-lysosomal, endocytic compartment in mDCs and further suggest that HIV-1 is able to selectively subvert components of the intracellular trafficking machinery required for formation of the DC-T-cell immunological synapse to facilitate its own cell-to-cell transfer and propagation. PMID- 15882446 TI - Brl1p -- a novel nuclear envelope protein required for nuclear transport. AB - In this article, we identify a cold-sensitive mutant of Xpo1p designated as xop1 2 (but will be referred to from here on as xpo1-ok) that is synthetically lethal with srm1-1, a Saccharomyces cerevisiae RCC1 homolog. xpo1-ok was a novel mutated allele with a single point mutation, T283P. Suppressors of xpo1-ok were isolated, and one of them was found to encode a novel nuclear envelope integral membrane protein designated as Brl1p (Brr6 like protein no. 1). Brl1p is homologous with Brr6p at the C-terminal domain, which is well conserved in the Brr6/Brl1 family. To characterize the function of Brl1p, a series of temperature-sensitive mutants of Brl1p were isolated. All of brl1 mutations were localized to the conserved C terminal domain that is essential for a function of Brl1p. Some brl1 alleles showed defects in nuclear export of either mRNA or protein, and nuclear pore clustering, similar to brr6-1. The cellular localization of Brl1p is also similar to that of Brr6p. The genetic analysis suggested that Brl1p functionally interacts with Brr6p. An interaction of Brl1p with Brr6p was shown by the two hybrid method. We hypothesize that Brl1p functions for nuclear export as a complex with Brr6p. PMID- 15882447 TI - Where do we go from here? Meeting Report on the Biophysical Society Discussion on 'Probing Membrane Microdomains', October 28-31, 2004, Asilomar, CA, USA. PMID- 15882448 TI - N-terminal fragment of B-type natriuretic peptide (NT-proBNP), a marker of cardiac safety during antipsychotic treatment. AB - BACKGROUND: The potential cardiotoxicity of antipsychotic drugs is well known. The N-terminal fragment of B-type natriuretic peptide (NT-proBNP) is considered to be a possible biomarker in clinical practice for the diagnosis and prognosis in patients with suspected heart failure. This pilot evaluation tests the influence of antipsychotic drugs on NT-proBNP concentration in view of the hypothesis that NT-proBNP could be used as marker for the tolerability and safety of antipsychotic medications. METHODS: On a routine basis, patient's blood samples were examined for NT-proBNP on days 0, 7 and 21 after initiation of a new antipsychotic monotherapy. All plasma samples were analysed for NT-proBNP using an electrochemiluminiscence immunoassay "ECLIA" (proBNP kit, Roche Diagnostics, Mannheim, Germany) on an Elecsys 2010 analyser. RESULTS: A difference was found in NT-proBNP values at day 0 between patients younger versus older than 40 years. Also women had comparatively lower NTproBNP on days 7 and 21. Smokers' levels of NT-proBNP values decreased more from day 0 to day 7. CONCLUSION: Our results suggest that antipsychotic medication influences the plasma concentration of NT proBNP, suggesting a possible method to identify high-risk-patients for cardiovascular adverse effects due to antipsychotic medication. Larger studies should further test this hypothesis. PMID- 15882449 TI - Predictive screening for regulators of conserved functional gene modules (gene batteries) in mammals. AB - BACKGROUND: The expression of gene batteries, genomic units of functionally linked genes which are activated by similar sets of cis- and trans-acting regulators, has been proposed as a major determinant of cell specialization in metazoans. We developed a predictive procedure to screen the mouse and human genomes and transcriptomes for cases of gene-battery-like regulation. RESULTS: In a screen that covered approximately 40 percent of all annotated protein-coding genes, we identified 21 co-expressed gene clusters with statistically supported sharing of cis-regulatory sequence elements. 66 predicted cases of over represented transcription factor binding motifs were validated against the literature and fell into three categories: (i) previously described cases of gene battery-like regulation, (ii) previously unreported cases of gene battery-like regulation with some support in a limited number of genes, and (iii) predicted cases that currently lack experimental support. The novel predictions include for example Sox 17 and RFX transcription factor binding sites that were detected in approximately 10% of all testis specific genes, and HNF-1 and 4 binding sites that were detected in approximately 30% of all kidney specific genes respectively. The results are publicly available at http://www.wlab.gu.se/lindahl/genebatteries. CONCLUSION: 21 co-expressed gene clusters were enriched for a total of 66 shared cis-regulatory sequence elements. A majority of these predictions represent novel cases of potential co-regulation of functionally coupled proteins. Critical technical parameters were evaluated, and the results and the methods provide a valuable resource for future experimental design. PMID- 15882450 TI - Atypical microbial infections of digestive tract may contribute to diarrhea in mucopolysaccharidosis patients: a MPS I case study. AB - BACKGROUND: Mucopolysaccharidoses are heritable, metabolic diseases caused by deficiency in an activity of one of specific lysosomal enzymes involved in degradation of mucoplysaccharides (glycosaminoglycans). Among many medical problems of patients with mucopolysaccharidoses, there are frequent episodes of diarrhea of unknown etiology. CASE PRESENTATION: A girl, diagnosed enzymatically for mucopolysaccharidosis type I (deficiency of alpha-L-iduronidase) at the age of 3 years and 9 months, was investigated until the age of 5 years and 4 months. Frequent loose stools and episodes of diarrhea, often accompanied by vomiting, were encountered. Detailed microbiological analyses were performed and atypical microbial infections (most often enetropathogenic Escherichia coli, but also other species, like Pseudomonas aeruginosa or Staphylococcus aureus, as well as adenoviruses) of the digestive tract were found in most severe diarrhea episodes. Often, isolations of pathogenic bacterial strains from stools of the investigated patient suffering from diarrhea were not obvious during the first screening, and only detailed microbiological studies, including re-isolation of colonies, gave the results of isolation of particular pathogenic strains (especially in the case of enetropathogenic E. coli). CONCLUSION: We conclude that atypical microbial infections of digestive tract may contribute significantly to diarrhea in mucopolysaccaridosis patients. Since isolated strains were not typical and their isolation was often possible only after detailed investigation (not during a standard screening), such atypical microbial infections of digestive tract of mucopolysaccharidosis patients could be usually overlooked to date. Importantly, these atypical infections could be effectively treated with antimicrobial agents. PMID- 15882451 TI - Erythroderma: a clinical study of 97 cases. AB - BACKGROUND: Erythroderma is a rare skin disorder that may be caused by a variety of underlying dermatoses, infections, systemic diseases and drugs. METHODS: We reviewed the clinical, laboratory and biopsy material of 97 patients diagnosed with erythroderma who were treated in our department over a 6-year period (1996 through 2002). RESULTS: The male-female ratio was 1.85:1. The mean age at diagnosis was 46.2 years. The most common causative factors were dermatoses (59.7%), followed by drug reactions (21.6%), malignancies (11.3%) and idiopathic causes (7.2%). Carbamazepine was the most common drug (57.1%). The best clinicopathologic correlation was found in cutaneous T-cell lymphoma and pityriasis rubra pilaris related erythroderma. Apart from scaling and erythema that were present in all patients, pruritus was the most common finding (97.5%), followed by fever (33.6%), lymphadenopathy (21.3%), edema (14.4%) and hyperkeratosis (7.2%). CONCLUSION: This study outlines that underlying etiologic factors of erythroderma may show geographic variations. Our series had a high percentage of erythroderma secondary to preexisting dermatoses and a low percentage of idiopathic cases. There was no HIV-infected patient among our series based on multiple serum antibody tests. The clinical features of erythroderma were identical, irrespective of the etiology. The onset of the disease was usually insidious except in drug-induced erythroderma, where it was acute. The group associated with the best prognosis was that related to drugs. PMID- 15882452 TI - Technology-dependency among patients discharged from a children's hospital: a retrospective cohort study. AB - BACKGROUND: Advances in medical technology may be increasing the population of children who are technology-dependent (TD). We assessed the proportion of children discharged from a children's hospital who are judged to be TD, and determined the most common devices and number of prescription medications at the time of discharge. METHODS: Chart review of 100 randomly selected patients from all services discharged from a children's hospital during the year 2000. Data were reviewed independently by 4 investigators who classified the cases as TD if the failure or withdrawal of the technology would likely have adverse health consequences sufficient to require hospitalization. Only those cases where 3 or 4 raters agreed were classified as TD. RESULTS: Among the 100 randomly sampled patients, the median age was 7 years (range: 1 day to 24 years old), 52% were male, 86% primarily spoke English, and 54% were privately insured. The median length of stay was 3 days (range: 1 to 103 days). No diagnosis accounted for more than 5% of cases. 41% were deemed to be technology dependent, with 20% dependent upon devices, 32% dependent upon medications, and 11% dependent upon both devices and medications. Devices at the time of discharge included gastrostomy and jejeunostomy tubes (10%), central venous catheters (7%), and tracheotomies (1%). The median number of prescription medications was 2 (range: 0-13), with 12% of cases having 5 or more medications. Home care services were planned for 7% of cases. CONCLUSION: Technology-dependency is common among children discharged from a children's hospital. PMID- 15882453 TI - Prevalence of oropharyngeal beta-lactamase-producing Capnocytophaga spp. in pediatric oncology patients over a ten-year period. AB - BACKGROUND: The aim of this study was to evaluate the prevalence of beta lactamase-producing Capnocytophaga isolates in young children hospitalized in the Pediatric Oncology Department of Hopital Sud (Rennes, France) over a ten-year period (1993-2002). METHODS: In neutropenic children, a periodic survey of the oral cavity allows a predictive evaluation of the risk of systemic infections by Capnocytophaga spp. In 449 children with cancer, 3,053 samples were collected by oral swabbing and plated on TBBP agar. The susceptibility of Capnocytophaga isolates to five beta-lactams was determined. RESULTS: A total of 440 strains of Capnocytophaga spp. were isolated, 309 (70%) of which were beta-lactamase producers. The beta-lactamase-producing strains were all resistant to cefazolin, 86% to amoxicillin, and 63% to ceftazidime. The proportion of strains resistant to third-generation cephalosporins remained high throughout the ten-year study, while susceptibility to imipenem and amoxicillin combined with clavulanic acid was always conserved. CONCLUSION: These results highlight the risk of antibiotic failure in Capnocytophaga infections and the importance of monitoring immunosuppressed patients and testing for antibiotic susceptibility and beta lactamase production. PMID- 15882454 TI - Dynamic simulation of red blood cell metabolism and its application to the analysis of a pathological condition. AB - BACKGROUND: Cell simulation, which aims to predict the complex and dynamic behavior of living cells, is becoming a valuable tool. In silico models of human red blood cell (RBC) metabolism have been developed by several laboratories. An RBC model using the E-Cell simulation system has been developed. This prototype model consists of three major metabolic pathways, namely, the glycolytic pathway, the pentose phosphate pathway and the nucleotide metabolic pathway. Like the previous model by Joshi and Palsson, it also models physical effects such as osmotic balance. This model was used here to reconstruct the pathology arising from hereditary glucose-6-phosphate dehydrogenase (G6PD) deficiency, which is the most common deficiency in human RBC. RESULTS: Since the prototype model could not reproduce the state of G6PD deficiency, the model was modified to include a pathway for de novo glutathione synthesis and a glutathione disulfide (GSSG) export system. The de novo glutathione (GSH) synthesis pathway was found to compensate partially for the lowered GSH concentrations resulting from G6PD deficiency, with the result that GSSG could be maintained at a very low concentration due to the active export system. CONCLUSION: The results of the simulation were consistent with the estimated situation of real G6PD-deficient cells. These results suggest that the de novo glutathione synthesis pathway and the GSSG export system play an important role in alleviating the consequences of G6PD deficiency. PMID- 15882455 TI - ABCC5, ERCC2, XPA and XRCC1 transcript abundance levels correlate with cisplatin chemoresistance in non-small cell lung cancer cell lines. AB - BACKGROUND: Although 40-50% of non-small cell lung cancer (NSCLC) tumors respond to cisplatin chemotherapy, there currently is no way to prospectively identify potential responders. The purpose of this study was to determine whether transcript abundance (TA) levels of twelve selected DNA repair or multi-drug resistance genes (LIG1, ERCC2, ERCC3, DDIT3, ABCC1, ABCC4, ABCC5, ABCC10, GTF2H2, XPA, XPC and XRCC1) were associated with cisplatin chemoresistance and could therefore contribute to the development of a predictive marker. Standardized RT (StaRT)-PCR, was employed to assess these genes in a set of NSCLC cell lines with a previously published range of sensitivity to cisplatin. Data were obtained in the form of target gene molecules relative to 106 beta-actin (ACTB) molecules. To cancel the effect of ACTB variation among the different cell lines individual gene expression values were incorporated into ratios of one gene to another. Each two-gene ratio was compared as a single variable to chemoresistance for each of eight NSCLC cell lines using multiple regression. In an effort to validate these results, six additional lines then were evaluated. RESULTS: Following validation, single variable models best correlated with chemoresistance (p < 0.001), were ERCC2/XPC, ABCC5/GTF2H2, ERCC2/GTF2H2, XPA/XPC and XRCC1/XPC. All single variable models were examined hierarchically to achieve two variable models. The two variable model with the highest correlation was (ABCC5/GTF2H2, ERCC2/GTF2H2) with an R2 value of 0.96 (p < 0.001). CONCLUSION: These results provide markers suitable for assessment of small fine needle aspirate biopsies in an effort to prospectively identify cisplatin resistant tumors. PMID- 15882456 TI - The antimicrobial susceptibility of Stenotrophomonas maltophilia isolates using three different methods and their genetic relatedness. AB - BACKGROUND: Stenotrophomonas maltophilia is inherently resistant to many antimicrobials. So far, antimicrobial susceptibility tests for S. maltophilia have not been fully standardized. The purpose of the study was to compare the susceptibility of S. maltophilia isolates against seven different antimicrobials using three different methods and to investigate their genetic relatedness. RESULTS: Although trimethoprim/sulfamethoxazole (SXT) and ciprofloxacin have the lowest MIC values, SXT (98.1%) and ticarcillin/clavulanate (TLc) (73.1%) were found to be the most effective antimicrobials by agar dilution method, which was in accordance with the breakpoints established by NCCLS. Disc diffusion and E test was in agreement with agar dilution method for SXT. When the isolation dates, clinics, antibiotyping, and AP-PCR data were investigated, two small outbreaks consisting of five and three cases were determined. CONCLUSION: By using the NCCLS criteria, disc diffusion and E-test were unreliable alternative methods for S. maltophilia, except for SXT. However, the significance of these data should be confirmed by further experimental and clinical studies. PMID- 15882457 TI - Role of cholesterol in parasitic infections. AB - The requirement of cholesterol for internalization of eukaryotic pathogens like protozoa (Leishmaniasis, Malaria and Toxoplasmosis) and the exchange of cholesterol along with other metabolites during reproduction in Schistosomes (helminths) under variable circumstances are poorly understood. In patients infected with some other helminthes, alterations in the lipid profile have been observed. Also, the mechanisms involved in lipid changes especially in membrane proteins related to parasite infections remain uncertain. Present review of literature shows that parasites induce significant changes in lipid parameters, as has been shown in the in vitro study where substitution of serum by lipid/cholesterol in medium and in experimental models (in vivo). Thus changes in lipid profile occur in patients having active infections with most of the parasites. Membrane proteins are probably involved in such reactions. All parasites may be metabolising cholesterol, but the exact relationship with pathogenic mechanism is not clear. So far, studies suggest that there may be some factors or enzymes, which allow the parasite to breakup and consume lipid/cholesterol. Further studies are needed for better understanding of the mechanisms involved in vivo. The present review analysis the various studies till date and the role of cholesterol in pathogenesis of different parasitic infections. PMID- 15882458 TI - Cardiac auscultation training of medical students: a comparison of electronic sensor-based and acoustic stethoscopes. AB - BACKGROUND: To determine whether the use of an electronic, sensor based stethoscope affects the cardiac auscultation skills of undergraduate medical students. METHODS: Forty eight third year medical students were randomized to use either an electronic stethoscope, or a conventional acoustic stethoscope during clinical auscultation training. After a training period of four months, cardiac auscultation skills were evaluated using four patients with different cardiac murmurs. Two experienced cardiologists determined correct answers. The students completed a questionnaire for each patient. The thirteen questions were weighted according to their relative importance, and a correct answer was credited from one to six points. RESULTS: No difference in mean score was found between the two groups (p = 0.65). Grading and characterisation of murmurs and, if present, report of non existing murmurs were also rated. None of these yielded any significant differences between the groups. CONCLUSION: Whether an electronic or a conventional stethoscope was used during training and testing did not affect the students' performance on a cardiac auscultation test. PMID- 15882459 TI - Serotypes, intimin variants and other virulence factors of eae positive Escherichia coli strains isolated from healthy cattle in Switzerland. Identification of a new intimin variant gene (eae-eta2). AB - BACKGROUND: Enteropathogenic Escherichia coli (EPEC) and Shigatoxin-producing Escherichia coli (STEC) share the ability to introduce attaching-and-effacing (A/E) lesions on intestinal cells. The genetic determinants for the production of A/E lesions are located on the locus of enterocyte effacement (LEE), a pathogenicity island that also contains the genes encoding intimin (eae). This study reports information on the occurrence of eae positive E. coli carried by healthy cattle at the point of slaughter, and on serotypes, intimin variants, and further virulence factors of isolated EPEC and STEC strains. RESULTS: Of 51 eae positive bovine E. coli strains, 59% were classified as EPEC and 41% as STEC. EPEC strains belonged to 18 O:H serotypes, six strains to typical EPEC serogroups. EPEC strains harbored a variety of intimin variants with eae-beta1 being most frequently found. Moreover, nine EPEC strains harbored astA (EAST1), seven bfpA (bundlin), and only one strain was positive for the EAF plasmid. We have identified a new intimin gene (eta2) in three bovine bfpA and astA-positive EPEC strains of serotype ONT:H45. STEC strains belonged to seven O:H serotypes with one serotype (O103:H2) accounting for 48% of the strains. The majority of bovine STEC strains (90%) belonged to five serotypes previously reported in association with hemolytic uremic syndrom (HUS), including one O157:H7 STEC strain. STEC strains harbored four intimin variants with eae-epsilon1 and eae gamma1 being most frequently found. Moreover, the majority of STEC strains carried only stx1 genes (13 strains), and was positive for ehxA (18 strains) encoding for Enterohemolysin. Four STEC strains showed a virulence pattern characteristic of highly virulent human strains (stx2 and eae positive). CONCLUSION: Our data confirm that ruminants are an important source of serologically and genetically diverse intimin-harboring E. coli strains. Moreover, cattle have not only to be considered as important asymptomatic carriers of O157 STEC but can also be a reservoir of EPEC and eae positive non O157 STEC, which are described in association with human diseases. PMID- 15882460 TI - Fractal ventilation enhances respiratory sinus arrhythmia. AB - BACKGROUND: Programming a mechanical ventilator with a biologically variable or fractal breathing pattern (an example of 1/f noise) improves gas exchange and respiratory mechanics. Here we show that fractal ventilation increases respiratory sinus arrhythmia (RSA) -- a mechanism known to improve ventilation/perfusion matching. METHODS: Pigs were anaesthetised with propofol/ketamine, paralysed with doxacurium, and ventilated in either control mode (CV) or in fractal mode (FV) at baseline and then following infusion of oleic acid to result in lung injury. RESULTS: Mean RSA and mean positive RSA were nearly double with FV, both at baseline and following oleic acid. At baseline, mean RSA = 18.6 msec with CV and 36.8 msec with FV (n = 10; p = 0.043); post oleic acid, mean RSA = 11.1 msec with CV and 21.8 msec with FV (n = 9, p = 0.028); at baseline, mean positive RSA = 20.8 msec with CV and 38.1 msec with FV (p = 0.047); post oleic acid, mean positive RSA = 13.2 msec with CV and 24.4 msec with FV (p = 0.026). Heart rate variability was also greater with FV. At baseline the coefficient of variation for heart rate was 2.2% during CV and 4.0% during FV. Following oleic acid the variation was 2.1 vs. 5.6% respectively. CONCLUSION: These findings suggest FV enhances physiological entrainment between respiratory, brain stem and cardiac nonlinear oscillators, further supporting the concept that RSA itself reflects cardiorespiratory interaction. In addition, these results provide another mechanism whereby FV may be superior to conventional CV. PMID- 15882461 TI - Comparison of array-based comparative genomic hybridization with gene expression based regional expression biases to identify genetic abnormalities in hepatocellular carcinoma. AB - BACKGROUND: Regional expression biases (REBs) are genetic intervals where gene expression is coordinately changed. For example, if a region of the genome is amplified, often the majority of genes that map within the amplified region show increased expression when compared to genes located in cytogenetically normal regions. As such, REBs have the potential to act as surrogates for cytogenetic data traditionally obtained using molecular technologies such as comparative genomic hybridization. However as REBs are identified using transcriptional information, detection of REBs may also identify local transcriptional abnormalities produced by both genetic and epigenetic mechanisms. RESULTS: REBs were identified from a set of hepatocellular carcinoma (HCC) gene expression profiles using a multiple span moving binomial test and compared to genetic abnormalities identified using array-based comparative genomic hybridization (aCGH). In the majority of cases, REBs overlapped genetic abnormalities as determined by aCGH. For example, both methods identified narrow regions of frequent amplification on chromosome 1p and narrow regions of frequent deletion on 17q. In a minority of cases, REBs were identified in regions not determined to be abnormal via other cytogenetic technologies. Specifically, expression biases reflective of cell proliferation were frequently identified on chromosome 6p21 23. CONCLUSION: Identification of REBs using a multiple span moving binomial test produced reasonable approximations of underlying cytogenetic abnormalities. However, caution should be used when attributing REBs identified on chromosome 6p to cytogenetic events in rapidly proliferating cells. PMID- 15882462 TI - Failure of a patient-centered intervention to substantially increase the identification and referral for-treatment of ambulatory emergency department patients with occult psychiatric conditions: a randomized trial [ISRCTN61514736]. AB - BACKGROUND: We previously demonstrated that a computerized psychiatric screening interview (the PRIME-MD) can be used in the Emergency Department (ED) waiting room to identify patients with mental illness. In that trial, however, informing the ED physician of the PRIME-MD results did not increase the frequency of psychiatric diagnosis, consultation or referral. We conducted this study to determine whether telling the patient and physician the PRIME-MD result would result in the majority of PRIME-MD-diagnosed patients being directed toward treatment for their mental illness. METHODS: In this single-site RCT, consenting patients with non-specific somatic chief complaints (e.g., fatigue, back pain, etc.) completed the computerized PRIME-MD in the waiting room and were randomly assigned to one of three groups: patient and physician told PRIME-MD results, patient told PRIME-MD results, and neither told PRIME-MD results.The main outcome measure was the percentage of patients with a PRIME-MD diagnosis who received a psychiatric consultation or referral from the ED. RESULTS: 183 (5% of all ED patients) were approached. 123 eligible patients consented to participate, completed the PRIME-MD and were randomized. 95 patients had outcomes recorded. 51 (54%) had a PRIME-MD diagnosis and 8 (16%) of them were given a psychiatric consultation or referral in the ED. While the frequency of consultation or referral increased as the intervention's intensity increased (tell neither = 11% (1/9), tell patient 15% (3/20), tell patient and physician 18% (4/22)), no group came close to the 50% threshold we sought. For this reason, we stopped the trial after an interim analysis. CONCLUSION: Patients willingly completed the PRIME-MD and 54% had a PRIME-MD diagnosis. Unfortunately, at our institution, informing the patient (and physician) of the PRIME-MD results infrequently led to the patient being directed toward care for their psychiatric condition. PMID- 15882463 TI - Hospital service areas -- a new tool for health care planning in Switzerland. AB - BACKGROUND: The description of patient travel patterns and variations in health care utilization may guide a sound health care planning process. In order to accurately describe these differences across regions with homogeneous populations, small area analysis (SAA) has proved as a valuable tool to create appropriate area models. This paper presents the methodology to create and characterize population-based hospital service areas (HSAs) for Switzerland. METHODS: We employed federal hospital discharge data to perform a patient origin study using small area analysis. Each of 605 residential regions was assigned to one of 215 hospital provider regions where the most frequent number of discharges took place. HSAs were characterized geographically, demographically, and through health utilization indices and rates that describe hospital use. We introduced novel planning variables extracted from the patient origin study and investigated relationships among health utilization indices and rates to understand patient travel patterns for hospital use. Results were visualized as maps in a geographic information system (GIS). RESULTS: We obtained 100 HSAs using a patient origin matrix containing over four million discharges. HSAs had diverse demographic and geographic characteristics. Urban HSAs had above average population sizes, while mountainous HSAs were scarcely populated but larger in size. We found higher localization of care in urban HSAs and in mountainous HSAs. Half of the Swiss population lives in service areas where 65% of hospital care is provided by local hospitals. CONCLUSION: Health utilization indices and rates demonstrated patient travel patterns that merit more detailed analyses in light of political, infrastructural and developmental determinants. HSAs and health utilization indices provide valuable information for health care planning. They will be used to study variation phenomena in Swiss health care. PMID- 15882464 TI - Clinical undergraduate training and assessment in primary health care: experiences gained from Crete, Greece. AB - BACKGROUND: Primary health care (PHC) is increasingly being introduced into undergraduate medical education. In Greece, the Faculty of Medicine of the University of Crete was the first to introduce a 4-week long training in primary health care. This paper presents the experiences gained from the initial implementation of the teaching of practice-based primary care in rural Crete and reports on the assessment scale that was developed. METHODS: 284 students' case write-ups from the 6 primary care units (PCUs) where they were allocated for the period 1990 to 1994 were analysed. The demographic data of the students and patients and the number of home visits were studied. Content analysis of the students' write-ups was carried out, using an assessment scale consisting of 10 dichotomous variables, in order to quantify eight (8) primary qualitative criteria. RESULTS: Internal reliability was estimated by the index KR20 = 0.67. Face and content validity was found to conform to the standards set for the course, while logistic linear regression analysis showed that the quality criteria could be used as an assessment scale. The number of home visits carried out varied between the various different PCUs (p < 0.001) and more were reported in the write-ups that fulfilled criteria related to the biopsychosocial approach (p < 0.05). Nine quantitative criteria were fulfilled in more than 90% of case reports, but laboratory investigations were reported only in 69.0% of case reports. Statistically significant differences between the PCUs were observed in the fulfilment of criteria related to the community approach, patient assessment and information related to the patient's perception of the illness, but not to those related to aspects of clinical patient management. Differences in reporting laboratory investigations (p < 0.001) are explained by the lack of such facilities in some PCUs. Demographic characteristics of the patients or the students' do not affect the criteria. CONCLUSION: The primary health care course achieved the objectives of introducing students to comprehensive, community oriented care, although there was variation between the PCUs. The assessment scale that was developed to analyse the case-write ups of the students provided data that can be used to evaluate the course. PMID- 15882465 TI - Mental health literacy in an educational elite -- an online survey among university students. AB - BACKGROUND: Mental health literacy is a prerequisite for early recognition and intervention in mental disorders. The aims of this paper are to determine whether a sample of university students recognise different symptoms of depression and schizophrenia and to reveal factors influencing correct recognition. METHODS: Bivariate and correspondence analyses of the results from an online survey among university students (n = 225). RESULTS: Most participants recognised the specific symptoms of depression. The symptoms of schizophrenia were acknowledged to a lower extent. Delusions of control and hallucinations of taste were not identified as symptoms of schizophrenia. Repeated revival of a trauma for depression and split personality for schizophrenia were frequently mistaken as symptoms of the respective disorders. Bivariate analyses demonstrated that previous interest in and a side job related to mental disorders, as well as previous personal treatment experience had a positive influence on symptom recognition. The correspondence analysis showed that male students of natural science, economics and philosophy are illiterate in recognising the symptoms depression and schizophrenia. CONCLUSION: Among the educational elite, a wide variability in mental health literacy was found. Therefore, it's important for public mental health interventions to focus on the different recognition rates in depression and schizophrenia. Possibilities for contact must be arranged according to interest and activity (e.g., at work). In order to improve mental health literacy, finally, education and/or internship should be integrated in high school or apprenticeship curricula. Special emphasis must be given towards the effects of gender and stereotypes held about mental illnesses. PMID- 15882466 TI - Discovery of a new human T-cell lymphotropic virus (HTLV-3) in Central Africa. AB - Human T-cell Leukemia virus type 1 (HTLV-1) and type 2 (HTLV-2) are pathogenic retroviruses that infect humans and cause severe hematological and neurological diseases. Both viruses have simian counterparts (STLV-1 and STLV-2). STLV-3 belongs to a third group of lymphotropic viruses which infect numerous African monkeys species. Among 240 Cameroonian plasma tested for the presence of HTLV-1 and/or HTLV-2 antibodies, 48 scored positive by immunofluorescence. Among those, 27 had indeterminate western-blot pattern. PCR amplification of pol and tax regions, using HTLV-1, -2 and STLV-3 highly conserved primers, demonstrated the presence of a new human retrovirus in one DNA sample. tax (180 bp) and pol (318 bp) phylogenetic analyses demonstrated the strong relationships between the novel human strain (Pyl43) and STLV-3 isolates from Cameroon. The virus, that we tentatively named HTLV-3, originated from a 62 years old Bakola Pygmy living in a remote settlement in the rain forest of Southern Cameroon. The plasma was reactive on MT2 cells but was negative on C19 cells. The HTLV 2.4 western-blot exhibited a strong reactivity to p19 and a faint one to MTA-1. On the INNO-LIA strip, it reacted faintly with the generic p19 (I/II), but strongly to the generic gp46 (I/II) and to the specific HTLV-2 gp46. The molecular relationships between Pyl43 and STLV-3 are thus not paralleled by the serological results, as most of the STLV-3 infected monkeys have an "HTLV-2 like" WB pattern. In the context of the multiple interspecies transmissions which occurred in the past, and led to the present-day distribution of the PTLV-1, it is thus very tempting to speculate that this newly discovered human retrovirus HTLV-3 might be widespread, at least in the African continent. PMID- 15882467 TI - Overweight and obesity at school entry among migrant and German children: a cross sectional study. AB - BACKGROUND: Overweight and obesity have become a global epidemic and are increasing rapidly in both childhood and adolescence. Obesity is linked both to socioeconomic status and to ethnicity among adults. It is unclear whether similar associations exist in childhood. The aim of the present study was to assess differences in overweight and obesity in migrant and German children at school entry. METHODS: The body mass index (BMI) was calculated for 525 children attending the 2002 compulsory pre-school medical examinations in 12 schools in Bielefeld, Germany. We applied international BMI cut off points for overweight and obesity by sex and age. The migration status of children was based on sociodemographic data obtained from parents who were interviewed separately. RESULTS: The overall prevalence of overweight in children aged 6-7 was 11.9% (overweight incl. obesity), the obesity prevalence was 2.5%. The prevalence of overweight and obesity was higher for migrant children (14.7% and 3.1%) than for German children (9.1% and 1.9%). When stratified by parental social status, migrant children had a significantly higher prevalence of overweight than German children in the highest social class. (27.6% vs. 10.0%, p = 0.032) Regression models including country/region and socioeconomic status as independent variables indicated similar results. The patterns of overweight among migrant children differed only slightly depending on duration of stay of their family in Germany. CONCLUSION: Our data indicate that children from ethnic minorities in Germany are more frequently overweight or obese than German children. Social class as well as family duration of stay after immigration influence the pattern of overweight and obesity in children at school entry. PMID- 15882468 TI - Electrical impedance along connective tissue planes associated with acupuncture meridians. AB - BACKGROUND: Acupuncture points and meridians are commonly believed to possess unique electrical properties. The experimental support for this claim is limited given the technical and methodological shortcomings of prior studies. Recent studies indicate a correspondence between acupuncture meridians and connective tissue planes. We hypothesized that segments of acupuncture meridians that are associated with loose connective tissue planes (between muscles or between muscle and bone) visible by ultrasound have greater electrical conductance (less electrical impedance) than non-meridian, parallel control segments. METHODS: We used a four-electrode method to measure the electrical impedance along segments of the Pericardium and Spleen meridians and corresponding parallel control segments in 23 human subjects. Meridian segments were determined by palpation and proportional measurements. Connective tissue planes underlying those segments were imaged with an ultrasound scanner. Along each meridian segment, four gold plated needles were inserted along a straight line and used as electrodes. A parallel series of four control needles were placed 0.8 cm medial to the meridian needles. For each set of four needles, a 3.3 kHz alternating (AC) constant amplitude current was introduced at three different amplitudes (20, 40, and 80 microAmps) to the outer two needles, while the voltage was measured between the inner two needles. Tissue impedance between the two inner needles was calculated based on Ohm's law (ratio of voltage to current intensity). RESULTS: At the Pericardium location, mean tissue impedance was significantly lower at meridian segments (70.4 +/- 5.7 Omega) compared with control segments (75.0 +/- 5.9 Omega) (p = 0.0003). At the Spleen location, mean impedance for meridian (67.8 +/- 6.8 Omega) and control segments (68.5 +/- 7.5 Omega) were not significantly different (p = 0.70). CONCLUSION: Tissue impedance was on average lower along the Pericardium meridian, but not along the Spleen meridian, compared with their respective controls. Ultrasound imaging of meridian and control segments suggested that contact of the needle with connective tissue may explain the decrease in electrical impedance noted at the Pericardium meridian. Further studies are needed to determine whether tissue impedance is lower in (1) connective tissue in general compared with muscle and (2) meridian-associated vs. non meridian-associated connective tissue. PMID- 15882469 TI - CYP3A4 and CYP3A5 genotyping by Pyrosequencing. AB - BACKGROUND: Human cytochrome P450 3A enzymes, particularly CYP3A4 and CYP3A5, play an important role in drug metabolism. CYP3A expression exhibits substantial interindividual variation, much of which may result from genetic variation. This study describes Pyrosequencing assays for key SNPs in CYP3A4 (CYP3A4*1B, CYP3A4*2, and CYP3A4*3) and CYP3A5 (CYP3A5*3C and CYP3A5*6). METHODS: Genotyping of 95 healthy European and 95 healthy African volunteers was performed using Pyrosequencing. Linkage disequilibrium, haplotype inference, Hardy-Weinberg equilibrium, and tag SNPs were also determined for these samples. RESULTS: CYP3A4*1B allele frequencies were 4% in Europeans and 82% in Africans. The CYP3A4*2 allele was found in neither population sample. CYP3A4*3 had an allele frequency of 2% in Europeans and 0% in Africans. The frequency of CYP3A5*3C was 94% in Europeans and 12% in Africans. No CYP3A5*6 variants were found in the European samples, but this allele had a frequency of 16% in the African samples. Allele frequencies and haplotypes show interethnic variation, highlighting the need to analyze clinically relevant SNPs and haplotypes in a variety of ethnic groups. CONCLUSION: Pyrosequencing is a versatile technique that could improve the efficiency of SNP analysis for pharmacogenomic research with the ultimate goal of pre-screening patients for individual therapy selection. PMID- 15882470 TI - Use of re-randomized data in meta-analysis. AB - BACKGROUND: Outcomes collected in randomized clinical trials are observations of random variables that should be independent and identically distributed. However, in some trials, the patients are randomized more than once thus violating both of these assumptions. The probability of an event is not always the same when a patient is re-randomized; there is probably a non-zero covariance coming from observations on the same patient. This is of particular importance to the meta analysts. METHODS: We developed a method to estimate the relative error in the risk differences with and without re-randomization of the patients. The relative error can be estimated by an expression depending on the percentage of the patients who were re-randomized, multipliers (how many times more likely it is to repeat an event) for the probability of reoccurrences, and the ratio of the total events reported and the initial number of patients entering the trial. RESULTS: We illustrate our methods using two randomized trials testing growth factors in febrile neutropenia. We showed that under some circumstances the relative error of taking into account re-randomized patients was sufficiently small to allow using the results in the meta-analysis. Our findings indicate that if the study in question is of similar size to other studies included in the meta-analysis, the error introduced by re-randomization will only minimally affect meta-analytic summary point estimate. We also show that in our model the risk ratio remains constant during the re-randomization, and therefore, if a meta-analyst is concerned about the effect of re-randomization on the meta-analysis, one way to sidestep the issue and still obtain reliable results is to use risk ratio as the measure of interest. CONCLUSION: Our method should be helpful in the understanding of the results of clinical trials and particularly helpful to the meta-analysts to assess if re-randomized patient data can be used in their analyses. PMID- 15882471 TI - Changes in the transcriptional profile of transporters in the intestine along the anterior-posterior and crypt-villus axes. AB - BACKGROUND: The purpose of this work was to characterize the expression of drug and nutrient carriers along the anterior-posterior and crypt-villus axes of the intestinal epithelium and to study the validity of utilizing whole gut tissue rather than purified epithelial cells to examine regional variations in gene expression. RESULTS: We have characterized the mRNA expression profiles of 76 % of all currently known transporters along the anterior-posterior axis of the gut. This is the first study to describe the expression profiles of the majority of all known transporters in the intestine. The expression profiles of transporters, as defined according to the Gene Ontology consortium, were measured in whole tissue of the murine duodenum, jejunum, ileum and colon using high-density microarrays. For nine transporters (Abca1, Abcc1, Abcc3, Abcg8, Slc10a2, Slc28a2, Slc2a1, Slc34a2 and Slc5a8), the mRNA profiles were further measured by RT-PCR in laser micro-dissected crypt and villus epithelial cells corresponding to the aforementioned intestinal regions. With respect to differentially regulated transporters, the colon had a distinct expression profile from small intestinal segments. The majority (59 % for p cutoff < or = 0.05) of transporter mRNA levels were constant across the intestinal sections studied. For the transporter subclass "carrier activity", which contains the majority of known carriers for biologically active compounds, a significant change (p < or = 0.05) along the anterior-posterior axis was observed. CONCLUSION: All nine transporters examined in laser-dissected material demonstrated good replication of the region-specific profiles revealed by microarray. Furthermore, we suggest that the distribution characteristics of Slc5a8 along the intestinal tract render it a suitable candidate carrier for monocarboxylate drugs in the posterior portion of the intestine. Our findings also predict that there is a significant difference in the absorption of carrier-mediated compounds in the different intestinal segments. The most pronounced differences can be expected between the adjoining segments ileum and colon, but the differences between the other adjoining segments are not negligible. Finally, for the examined genes, profiles measured in whole intestinal tissue extracts are representative of epithelial cell-only gene expression. PMID- 15882472 TI - The Bhopal disaster and its aftermath: a review. AB - On December 3 1984, more than 40 tons of methyl isocyanate gas leaked from a pesticide plant in Bhopal, India, immediately killing at least 3,800 people and causing significant morbidity and premature death for many thousands more. The company involved in what became the worst industrial accident in history immediately tried to dissociate itself from legal responsibility. Eventually it reached a settlement with the Indian Government through mediation of that country's Supreme Court and accepted moral responsibility. It paid $470 million in compensation, a relatively small amount of based on significant underestimations of the long-term health consequences of exposure and the number of people exposed. The disaster indicated a need for enforceable international standards for environmental safety, preventative strategies to avoid similar accidents and industrial disaster preparedness. Since the disaster, India has experienced rapid industrialization. While some positive changes in government policy and behavior of a few industries have taken place, major threats to the environment from rapid and poorly regulated industrial growth remain. Widespread environmental degradation with significant adverse human health consequences continues to occur throughout India. PMID- 15882474 TI - Is there a role for less invasive therapeutic approaches for low- and intermediate-grade organ-confined prostate cancer? PMID- 15882473 TI - Protection of cortical cells by equine estrogens against glutamate-induced excitotoxicity is mediated through a calcium independent mechanism. AB - BACKGROUND: High concentrations of glutamate can accumulate in the brain and may be involved in the pathogenesis of neurodegenerative disorders such as Alzheimer's disease. This form of neurotoxicity involves changes in the regulation of cellular calcium (Ca2+) and generation of free radicals such as peroxynitrite (ONOO-). Estrogen may protect against glutamate-induced cell death by reducing the excitotoxic Ca2+ influx associated with glutamate excitotoxicity. In this study, the inhibition of N-methyl-D-aspartate (NMDA) receptor and nitric oxide synthase (NOS) along with the effect of 17beta-estradiol (17beta-E2) and a more potent antioxidant Delta8, 17beta-estradiol (Delta8, 17beta-E2) on cell viability and intracellular Ca2+ ([Ca2+]i), following treatment of rat cortical cells with glutamate, was investigated. RESULTS: Primary rat cortical cells were cultured for 7-12 days in Neurobasal medium containing B27 supplements. Addition of glutamate (200 microM) decreased cell viability to 51.3 +/- 0.7% compared to control. Treatment with the noncompetitive NMDAR antagonist, MK-801, and the NOS inhibitor, L-NAME, completely prevented cell death. Pretreatment (24 hrs) with 17beta-E2 and Delta8, 17beta-E2 (0.01 to 10 microM) significantly reduced cell death. 17beta-E2 was more potent than Delta8, 17beta-E2. Glutamate caused a rapid 2.5 fold increase in [Ca2+]i. Treatment with 0.001 to 10 microM MK-801 reduced the initial Ca2+ influx by 14-41% and increased cell viability significantly. Pretreatment with 17beta-E2 and Delta8, 17beta-E2 had no effect on Ca2+ influx but protected the cortical cells against glutamate-induced cell death. CONCLUSION: Glutamate-induced cell death in cortical cultures can occur through NMDAR and NOS-linked mechanisms by increasing nitric oxide and ONOO-. Equine estrogens: 17beta-E2 and Delta8, 17beta-E2, significantly protected cortical cells against glutamate-induced excitotoxicity by a mechanism that appears to be independent of Ca2+ influx. To our knowledge, this is a first such observation. Whether the decrease in NOS related products such as ONOO-, is a mechanism by which estrogens protect against glutamate toxicity, remains to be investigated. Estrogen replacement therapy in healthy and young postmenopausal women may protect against neurodegenerative diseases by these mechanisms. PMID- 15882475 TI - Highlights from the 2005 American Society of Clinical Oncology Prostate Cancer Symposium, Orlando, Florida. PMID- 15882476 TI - Role of 5a-reductase inhibitors and selective estrogen receptor modulators as potential chemopreventive agents for prostate cancer. PMID- 15882477 TI - Combined androgen blockade: the case for bicalutamide. AB - Combination therapy consists of castration plus an antiandrogen. Following medical or surgical castration, the androgen receptor can be activated by adrenal androgens, low levels of residual testosterone, and ligand-independent activators. The survival benefit of combination therapy compared with castration alone is one of the most studied questions in urology. Results from trials comparing combination therapy to castration alone are variable. A metaanalysis of 26 randomized trials indicated that the type of antiandrogen used is relevant. Combination therapy using nonsteroidal antiandrogens was associated with a statistically significant overall survival benefit. In contrast, combination therapy using steroidal antiandrogens was associated with reduced survival compared with castration alone. Bicalutamide 50 mg has a number of advantages compared with nilutamide and flutamide when used in combination with castration. These include an improved side-effect profile, once-daily dosing, more potent inhibition of androgen-independent activation of the androgen receptor through favorable interactions with nuclear coactivators and corepressors, and evidence for improved survival in one randomized trial. An analysis combining historic trial data suggests that bicalutamide 50 mg in addition to androgen deprivation may reduce the hazard ratio (HR) for prostate cancer mortality by 20% (HR, 0.80; 95% CI, 0.66-0.98). PMID- 15882478 TI - Genetic alterations in prostate cancer. AB - Prostate cancer is the most common nondermatologic malignancy in men. Prostate cancer is characterized by clinical and biologic heterogeneity that has complicated molecular and epidemiologic studies. Like other epithelial malignancies, prostate tumors exhibit complex karyotypic abnormalities and harbor many specific genetic alterations. Although recent work has begun to elucidate many of the specific mutations associated with prostate cancer, we still lack a clear understanding of the complement of genetic changes that suffice to program the malignant state. Here, we review our current understanding of the genetic changes found in prostate cancer and explore the connections between specific genetic alterations and malignant phenotypes including cell growth, survival, invasion, and metastasis. PMID- 15882479 TI - Update on fused capromab pendetide imaging of prostate cancer. AB - The primary objective of this overview is to apprise clinical urologists and oncologists of the current state of fused multimodality imaging of prostate cancer, which can be applied to optimize treatment by ensuring that a patient's disease is characterized as well as current imaging technology permits. The focus of this study is the monoclonal antibody capromab pendetide, which targets prostate specific membrane antigen (PSMA), a type II membrane glycoprotein strongly associated with prostate cancer. Identifying where capromab pendetide uptake occurs can be done accurately if this functional imaging modality is combined with a modality that provides anatomic detail, such as computed tomography (CT) or magnetic resonance imaging (MRI). Image fusion, or coregistration, which is overlaying the functional images of capromab pendetide uptake on the anatomic CT or MRI images, provides a detailed map of cancer localization inside and outside the prostate gland. This same principle of fusing functional images on anatomic images is the basis for enormous growth of positron emission tomography with CT during the past 2 years. Positron emission tomography imaging has a different functionality base than does capromab pendetide, and thus the 2 modalities should be complementary. However, the key to both functional imaging modalities is accurate fusion with anatomic images, which is illustrated in our case reports. The cases cited demonstrate the need to optimize every phase of imaging from patient preparation to reading and reporting increased PSMA concentration seen on the fused images. Reference is also made to applying capromab pendetide/CT fused imaging to radiation therapy planning. PMID- 15882480 TI - Lifestyle correlates of health perception and treatment satisfaction in a clinical cohort of men with prostate cancer. AB - There is a growing need to understand how nutritional and lifestyle practices may optimize quality of life (QOL) and health after diagnosis for the 1.5 million men living with prostate cancer in the United States. We are establishing a clinical cohort of men with prostate cancer at the University of California San Francisco. Men completed detailed dietary and lifestyle questionnaires annually and provided consent for blood and tissue specimens to be stored for research if they underwent radical prostatectomy. We examined the feasibility of establishing this cohort and analyzepreliminary baseline data on participant demographics, lifestyle habits, and QOL using c2 and t-tests and logistic regression models. Between February 2002 and July 2004, we enrolled 343 men with prostate cancer into the survey portion of this cohort. The response rate was approximately 85% via in-clinic enrollment and 30% via mail enrollment. Based on analysis of the first 193 men enrolled, there was a high level of treatment satisfaction in this population (88% of men were satisfied or extremely satisfied with treatment) and positive reports of general health perception (73% of men perceived themselves to be in excellent [34%] or very good [39%] health). Whether treatment interfered with diet was an independent predictor of health perception and treatment satisfaction. Use of dietary supplements was high (90%) in this well-educated population. In conclusion, we demonstrated good feasibility for conducting this longitudinal study and observed initial indications that diet and other lifestyle practices were important predictors of patient QOL. PMID- 15882481 TI - Web-based intervention for changing attitudes of obesity among current and future teachers. AB - OBJECTIVE: To evaluate attitude change among student teachers and schoolteachers when exposed to a Web-based educational module promoting size acceptance. DESIGN: Subjects were randomly assigned to a control group or 1 of 4 treatment groups to evaluate the effect of module, presenter credibility, and/or image on attitudes of obesity and processes of change using the Elaboration Likelihood Model (ELM) testing cognitive and psychological cues. On-line assessment occurred at the pretest, posttest, and 6-week follow-up. SETTING: Web-based. PARTICIPANTS: 258 adults, mean age 26.8 +/- 10.2 years. INTERVENTION: Module content included factors related to obesity, implications of weight loss efforts, classroom activities, and bias-free intervention techniques. MAIN OUTCOME MEASURE(S): Attitudes of obesity by the Anti-Fat Attitudes Test and the effect of the presenter using a bipolar rating scale. ANALYSIS: One-way repeated measures analysis of variance, analysis of covariance, and general post hoc analysis. RESULTS: Negative attitudes decreased in treatment groups between pretest to posttest (P < .001) and pretest to follow-up (P < or = .006). Unlike the credible "nonfat" presenter, exposure to the credible "fat" presenter positively influenced attitude change (P = .031). CONCLUSIONS AND IMPLICATIONS: On-line communication of size acceptance improved attitudes of obesity long term. The findings support the ELM. The module demonstrates promise for teacher training in size sensitivity. PMID- 15882482 TI - New moccasins: articulating research approaches through interviews with faculty and staff at Native and non-Native academic institutions. AB - OBJECTIVE: The goals of this study were to explore the perceptions of professionals concerning research, its different approaches, and appropriate future directions with Native American communities, particularly in relation to nutrition issues. DESIGN: Semistructured qualitative interviews. setting: Interviews were conducted at Native and non-Native academic institutions, at other relevant locations, and over the telephone. PARTICIPANTS: Participants (N = 20) were from Native and non-Native academic institutions and had experience working with research in Native American communities. PHENOMENA OF INTEREST: Relationships between Native and non-Native academic institutions, worldviews regarding research and American Indian communities, and beneficial research within American Indian communities. ANALYSIS: Interviews were audiotaped, transcribed verbatim, and returned to the participants for review. Analysis of interview material involved eliciting themes. RESULTS: Some participants acknowledged different cultural worldviews in relation to research. Many participants provided insight on how to define beneficial research. Most said building trust between Native and non-Native academic institutions is an important step in developing effective research relationships. CONCLUSIONS AND IMPLICATIONS: These findings are a first step toward creating a more equitable process of research that acknowledges different cultural worldviews and values community involvement within Native American communities. PMID- 15882483 TI - Qualitative investigation of differences in benefits and challenges of eating fruits versus vegetables as perceived by Canadian women. AB - OBJECTIVE: This study examined the perceived differences in benefits and challenges relating to fruit versus vegetable consumption among a purposive, convenience sample of Canadian women. DESIGN: This inductive, qualitative study involved 8 semistructured group interviews conducted by an experienced moderator. SETTING: Interviews were conducted at public health units in southern Ontario. PARTICIPANTS: Forty-seven women, aged 20 to 44 years, were recruited through existing community programs and newspaper advertisements. ANALYSIS: The constant comparison method of data analysis was used to identify overarching themes. RESULTS: Five themes were identified: (1) fruits "fill the gap between meals" (the main benefit); (2) concern about "pesticides and parasites and bacteria"; and (3) "How can something look so good and have no taste?" (main challenges of eating fruit); (4) vegetables make meals "appealing" (main benefit); and (5) the "social" dimension of eating vegetables (main challenge). CONCLUSIONS AND IMPLICATIONS: Participants readily described different benefits and challenges relating to consumption of fruits versus vegetables. Tailored nutrition messages addressing perceived differences in the benefits and challenges for eating fruits versus vegetables may be needed to encourage increased consumption of these foods. Further research can determine whether these perceptions are widely held. PMID- 15882484 TI - Overweight consumers' salient beliefs on omega-3-enriched functional foods in Australia's Illawarra region. AB - OBJECTIVE: To determine consumer salient beliefs toward functional foods enriched with omega-3 fatty acids. DESIGN: Focus group interviews with adult consumers using the Theory of Planned Behavior (TPB) as a theoretical framework. SETTING: Community-based residents living in the Illawarra region of New South Wales, Australia. PARTICIPANTS: Forty-two overweight participants (29 female, 13 male) aged 30 to 80 years recruited by advertisement and attending 1 of 6 focus groups, which were recorded and transcribed verbatim. ANALYSIS: Content analysis was carried out, and subcategories were developed to capture the emerging themes according to the TPB model. RESULTS: Most participants were aware of a range of potential benefits of omega-3 fatty acids, but they had reservations about the ability of omega-3-enriched foods to deliver a health benefit. They were concerned about whether these foods were labeled clearly and about the possibility of overdosing. Family and friends were viewed as important in introducing participants to novel foods on the market. Participants regarded dietitians as a credible source and were least trusting of food companies and scientists. Overall, participants reported that cost was a major barrier, and that they would not necessarily trade taste for health benefits. Adding omega-3 fatty acids to foods regarded as less healthful was viewed more as a gimmick rather than a real health benefit. CONCLUSIONS AND IMPLICATIONS: The consumer attitudes and purchase intentions identified in this study will be helpful to educators as they plan messages and strategies to guide dietary choices related to products enriched with omega-3 fatty acids. PMID- 15882485 TI - Nutrition education, behavioral theories, and the scientific method: another viewpoint. AB - In a previous viewpoint in this journal, David Buchanan suggested that the scientific model is inappropriate, ineffective, and possibly unethical in nutrition education research and championed the use of a more humanistic model. The purpose of this article is to provide a counterpoint to Buchanan's views. More specifically, this article will provide another interpretation of the current status of the scientific method in the social and behavioral sciences, will examine how the field of nutrition education has benefited from the use of the scientific method to evaluate behavioral theories, and will discuss what we are trying to achieve by using theory in nutrition education. PMID- 15882486 TI - Pizza Please: an interactive nutrition evaluation for second and third grade students. PMID- 15882487 TI - Nutrition Education Aimed at Toddlers (NEAT) curriculum. PMID- 15882488 TI - Using media messaging to promote healthful eating and physical activity among urban youth. PMID- 15882489 TI - Use of films to teach critical thinking. PMID- 15882490 TI - Bringing you fresh food from local farms and our garden: a college class designs a program to meet peer and institutional needs. PMID- 15882491 TI - Cooking classes increase fruit and vegetable intake and food safety behaviors in youth and adults. PMID- 15882492 TI - [Policies of prevention and monitoring of tobacco dependency]. PMID- 15882493 TI - [Complementary or alternative medicine? A dilemma for the public health system?]. PMID- 15882494 TI - [The Framingham function overestimates stroke risk for diabetes and metabolic syndrome among Spanish population]. AB - OBJECTIVE: To estimate stroke risk for diabetes, isolated or associated to metabolic syndrome (MS) according WHO and National Cholesterol Education Program (NCEP) criteria. DESIGN: Multicentre and prospective cohort study. SETTING: Primary health care. PARTICIPANTS: Subjects between 55-85 years-old without any evidence of stroke, included from 1998 in a random population sample for MS follow-up during routine practice in Reus (Tarragona, Spain). MAIN MEASUREMENTS: Stroke risk was achieved using Framingham function by means of a computerized algorithm using a diagnostic factorial design (diabetes and/or MS). Theoretical stroke risk and cumulated incidence of stroke events (1998-2003) were compared. RESULTS: Among 728 subjects (412 women, mean age =66 years old, body mass index =29 kg/m2), 457 (62.8%) did not have diabetes, nor MS, 93 (12.8%) had MS without diabetes, 72 (9.9%) diabetes without MS, and 106 (14.5%) presented both conditions (WHO rules). According NCEP criteria were 60.7%, 14.8%, 7.8%, and 16.7%, respectively. Ten-year estimated stroke risk accounted for (WHO/NCEP) 8.4/9.1%, 10.8/10.5%, 18/17.3%, and 18.8/19.1%. Cumulated incidence for stroke events were: 2.8%, 1.4%, 5.4%, and 3.8% (WHO), and 2.5%, 2.8%, 3.5%, and 5.8%, respectively (NCEP). CONCLUSIONS: Stroke risk scores were extremely increased among diabetic subjects irrespective to MS diagnose. The Framingham function probably overestimates stroke risk among Spanish individuals. PMID- 15882495 TI - [Framingham is in Massachusetts]. PMID- 15882496 TI - [Gender and psychiatric drugs: a qualitative study to find the views of prescription-fillers]. AB - OBJECTIVES: To find the views of those filling out prescriptions on the factors that affect differential prescription of psychiatric medication, and on reducing the prescription; and to analyse their discourse for possible implicit gender bias. DESIGN: Qualitative methodology. Semi-structured interviews conducted in 2002-2003. SETTING: Urban and rural health centres. PARTICIPANTS: Family and community medicine and psychiatry professionals working in the public health services of Andalusia, the Community of Madrid, and the Basque Country. Segmentation criteria: community of residence, age, sex, medical speciality, and location of centre. METHODS: Structural sample. Interviews conducted by the research team. Strategy of analysis in two distinct processes: contents analysis with agreed categories and gender analysis of the discourse. RESULTS. The prescribers found that taking psychiatric medication was associated with sex, age, social and economic category, and social expectations. In the discourses we found consumer profiles emerging that were not found in the epidemiological literature. Discourse analysis showed implicit gender bias in the beliefs of some prescribers. CONCLUSIONS. To minimise the growing offer and demand for psychiatric medication, prescribers thought training in diagnosis and psychotherapy should be improved, case-loads should be reduced, co-ordination between services improved and alternative treatments favoured. To minimise gender bias in the diagnosis and prescription of psychiatric medication, we think training is needed on the influence of cultural and gender factors on the process of construction of identities. PMID- 15882497 TI - [Sleeping habits and sleep disorders during adolescence: relation to school performance]. AB - OBJECTIVE: To determine the prevalence of sleep disorders in adolescence. To describe sleeping habits of adolescents in relation to sleep disorders and associated factors. To determine the relation between sleep disorders/inappropiate sleeping habits and school performance. DESIGN: Observational, descriptive, cross-sectional study. SETTING: Secondary school of Cuenca (city in Spain). PARTICIPANTS: 1293 school children of first and fourth curses of secondary education. MAIN MEASURES: Structured questionnaire with opened and closed questions on sleeping habits during weekdays and at weekends and sleep disorders to be answered by the adolescents anonymously and on their own. Student's school performance with relation with to sleeping habits and sleep disorders were determined. RESULTS: 1155 students out of 1293 (response rate 89.33%) answered the questionnaire, 537 (45.9%) boys and 618 (54.1%) girls, 14 years old on average (between 11-18 years). On weekdays students went to bed at 23.17 h and got up at 7.46 h (average sleeping time =8 hours and 18 minutes). At weekends they went to bed at 1.02 h and got up at 10.42 h (average sleeping time =9 hours and 40 minutes). 45.4% of students said to sleep badly on Sunday night's. On average the number of subjects failed in class is higher with adolescents who complain about sleep (2.28 vs 1.91; P=.04), who are tired at waking up time (2.17 vs 1.97; P=.048) and who have morning sleepiness (2.17 vs 1.75; P=.004). CONCLUSIONS: Schools hours cause deficit sleeping time during weekdays which is partly made up for at weekend. At weekends there is an interruption of the adolescent's sleeping habits. School performance of adolescents with sleep disorders is lower. PMID- 15882498 TI - [Mobile telephones: playing with our health?]. PMID- 15882499 TI - [Primary care doctors needs for information: analysis of questions and how they were answered]. AB - OBJECTIVE: To assess what information primary care doctors from 5 health areas of Madrid need, through the description of the frequency and kind of clinical questions that arise, the pattern of search for the answers and how well they were answered. DESIGN: Descriptive, observational study. Setting. Primary care consultations in health areas 1, 3, 8, 9, and 10, Madrid, Spain. PARTICIPANTS: Randomised sample, stratified by zone, and kind of doctor, of 125 doctors from each area (Madrid). Residents, locums, and university teachers were excluded. INTERVENTIONS: Doctors will be invited to be observed by video camera for about four hours of on-demand consultation. They will be asked between patients to specify the questions that arise and the sources of information used in the consultation. Unresolved questions will be followed up by means of phone contact 2 weeks later, to see whether they had found answers and what methods they used to find them. MAIN MEASUREMENTS: MAIN VARIABLES: number of recognised clinical questions formulated and their answers; number of clinical questions not recognised (as observed by video); each question's theme and kind of information; time used to look for answers; information resources used. Other variables: characteristics of the PC professionals taking part, consultations, health centres, and case-loads. DISCUSSION: Restrictions: Hawthorne effect or bias in the person observed and participants stage fright. Applicability: the results of the research will be used to plan the resources needed for doctors to enjoy and put into practice the best scientific knowledge. PMID- 15882500 TI - [Controversies in auricular fibrillation: does sinus rhythm have to be maintained?]. PMID- 15882501 TI - [Alcoholic detoxification in primary care]. PMID- 15882502 TI - [Prevalence of metabolic syndrome in patients of an urban primary care centre]. PMID- 15882503 TI - [Use of ARA II and Scientific Evidence]. PMID- 15882504 TI - [Acute pancreatitis due to lipid-lowering drugs]. PMID- 15882505 TI - [Osteoporosis and calcaneus ultrasound. Pharmaceutical businesses and primary care. What are we doing?]. PMID- 15882506 TI - [Prevalence of carbohydrate disorders in primary care]. PMID- 15882509 TI - [Prevalence of an abnormal ankle-brachial index in relation to the cardiovascular risk estimated by the Framingham function]. AB - BACKGROUND AND OBJECTIVE: The measurement of the ankle-brachial index (ABI) is a straightforward method for the detection of atherosclerosis in the lower limbs. An abnormal ABI (< 0.9 or > 1.4) is associated with the development of cardiovascular disease and cardiovascular and all-cause mortality. Despite this, its measurement in clinical practice is underused. The objective of the present study was to evaluate the relation of the ABI with the cardiovascular risk determined by traditional risk functions in a population in primary prevention. PATIENTS AND METHOD: 1001 subjects without known cardiovascular disease attended in primary care were invited to participate in the study. Cardiovascular risk and ABI measurements were calculated in all participants. RESULTS: A low (< 0.9) ABI was found in a 3.8% of the participants, 3.9% females and 3.6% males. An abnormal ABI (< 0.9 or > 1.4) was found in 6.4% of all subjects, 5.2% females and 8.8% males. In a multivariable analysis age (OR = 1.09 for each year; 95% CI 1.03 1.15), smoking habit (OR = 2.96; 95% CI 1.51-5.80), HDL-cholesterol levels (OR = 0.98 for each mg/dl; 95% CI, 0.95-0.99) and hypertension (OR = 1.80; 95% CI, 1.05 3.06) were related with an abnormal ABI. Subjects were divided according to their risk stratification. The percentage of low, moderate and high risk individuals with an abnormal ABI was 2.6%, 8.7% and 14.9% respectively. CONCLUSIONS: In primary prevention, one in ten individuals with moderate risk and one in six individuals with high risk have an abnormal ABI. In these subjects there is an indication for intensive preventive strategies and antiagregation. PMID- 15882510 TI - [Study of deferoxamine in subcutaneous profusion treatment of iron overload in myelodysplastic syndromes]. AB - BACKGROUND AND OBJECTIVE: The progressive accumulation of iron in the organism contribute to one of the most important problems of morbidity and mortality in patients with myelodysplastic syndrome (MDS). We present an observational protocol, multicentre, open and non-aleatheorised, in patients diagnosed with MDS on transfusional regime with iron overload. The study was meant to prove the effectiveness of the parenteral treatment with desferrioxamine using continuous devices of subcutaneous profusion and evaluate the evolution of iron overload as well as transfusional requirements. PATIENTS AND METHOD: There were 28 patients (12 men and 16 women), 12 AR, 15 AS and 1 unknown. Average monitoring lasted 13.5 months and there was a gradual loss of patients. 11 of them had gone during the first year. RESULTS: After 12 months the average of ferritin decreased by 258.51 ng/dl (DE 1208.04; p = 9.4) and after 24 months, it decreased by 979.6 ng/dl (DE 810.31; p = 0.1). After 12 months the average of requirements increased by 60.57 gHb/month (DE 183.7; p = 0.029) and after 24 months, it increased by 167.3 g/Hb/month (DE 406.5; p = 0.36). CONCLUSIONS: Desferroxiamine treatment is effective at least to prevent an iron overload in these patients, and therefore should be incorporated in the clinical practice. PMID- 15882511 TI - [Factors predicting a short length of stay for acute exacerbations of chronic obstructive pulmonary disease]. AB - BACKGROUND AND OBJECTIVE: We intended to identify the factors predicting a short length of stay in patients with chronic obstructive pulmonary disease (COPD) and acute exacerbations in order to select those who are tributary to be admitted in a short stay unit. PATIENTS AND METHOD: Several clinical and biochemical characteristics were compared in a group of patients with COPD admitted in a short stay unit during an acute exacerbation, classified as lasting 4 days or > 4 days of hospital stay. RESULTS: A comorbidity index of 2 [1-2] vs 3 [2-3.5], a number of breaths per minute of 29.0 (6.5) vs 33.6 (6.8) and a pCO2 value on admission of 39.1 (5.3) vs 50.7 (12.0) mmHg were factors independently associated with a shorter stay (p < 0.001 in all). CONCLUSIONS: Some clinical characteristics of COPD patients are predictive of a short length of stay during acute exacerbations. PMID- 15882512 TI - [Short Zarit scale in palliative care]. AB - BACKGROUND AND OBJECTIVE: We aimed to reduce the Zarit scale (ZS) for the assessment of caregiver burden in palliative care. PATIENTS AND METHOD: ZS was performed in 61 caregivers of palliative care patients admitted to hospital. The field explored by each item was decided by an expert committee and a logistic regression and ROC curve were performed. RESULTS: ZS reduction to 7 items was achieved with a sensitivity and specificity of 100%. CONCLUSIONS: A reduced Zarit scale is useful to identify caregiver burden in palliative care. It is also easy to use. PMID- 15882513 TI - [Iron overload in myelodysplastic syndromes. Role of iron chelators]. PMID- 15882514 TI - [Preventive medicine and the catastrophic failures of public health: we fail because we are late]. PMID- 15882515 TI - [Therapeutic strategies in HIV infection]. PMID- 15882516 TI - [Pathogenesis of primary nonalcoholic fatty liver disease]. AB - The challenges of growing prevalence and evident trend to progressive damage of primary nonalcoholic fatty liver disease confront a poorly understood pathogenesis. It appears to develop in two steps. First, a high adipocyte protein production in the context of a silent inflammatory background causes insulin resistance in adipose tissue. It leads both to lipolysis, with increase of the circulating and hepatic uptake of free fatty acids, and hyperinsulinemia. Within hepatocytes, the subsequent lipogenesis, together with a decreased secretion of lipoproteins, induces an accumulation of excessive hepatic triglycerides (steatosis), impliying some oxidative damage, but it remain balanced by uncoupling protein upregulation and antioxidant systems activation. Second, a more forceful fat catabolism by beta and omega oxidation results in respiratory chain hyperactivity with overproduction of free radicals and reactive oxygen species that exceed the antioxidant capacity. These agents lead to hepatocellular injury and necrosis, inflammatory infiltration and fibrosis (steatohepatitis) through induction of Fas ligand and cytokines (tumor necrosis factor alpha, transforming growth factor beta, interleukin-8), and lipid peroxidation and by products (malondialdehyde and 4-hydroxynonenal). Other mechanisms (hepatic iron, Kupffer cells dysfunction or endotoxemia) play uncertain roles. PMID- 15882517 TI - [Heparin-induced thrombocytopenia with thrombosis and cardiovascular surgery]. PMID- 15882518 TI - [Pneumatosis cystoides intestinalis and irritable bowel syndrome]. PMID- 15882519 TI - [Autologous transfusion in a pregnant woman with anti-U and anti-He (Henshaw) antibodies]. PMID- 15882521 TI - The article by Wilde et al. in Vaccine could contain some important factual errors. PMID- 15882522 TI - Candidate multi-peptide-vaccine against classical swine fever virus induced potent immunity with serological marker. AB - Our previous study proposed a protective multi-peptide-vaccine (MPV) with Freund's adjuvant against classical swine fever virus (CSFV). In this study, another candidate MPV, using aluminum adjuvant, was further examined. All immunized pigs kept healthy during the experimental period, while the control group rapidly showed clinical symptoms and died. Moreover, anti-sera from MPV immunized pigs could interact with peptides involved in the MPV, in contrast to anti-sera from non-immunized or infected ones. This property permits MPV immunized pigs to be easily differentiated from infected ones with simple serological method. Therefore, this new MPV is suitable to act as a candidate marker vaccine against CSFV. PMID- 15882523 TI - Immune responses induced by lower airway mucosal immunisation with a human papillomavirus type 16 virus-like particle vaccine. AB - Cervical cancer results from cervical infection by human papillomaviruses (HPV), especially HPV16. Previous studies have shown that intramuscular vaccination of women with an HPV16 virus-like particle (VLP) vaccine induced a strong IgG response and protected against genital HPV16 infection. However, an alternative route of administration that avoids parenteral injection while inducing mucosal immunity might facilitate vaccine implementation in some settings, and partially overcome the substantial variation in HPV16 antibodies at the cervix seen in ovulating women. In this study, women were vaccinated with escalating doses of HPV16L1 VLPs via nasal nebulisation, bronchial aerosolisation, or a combination of intramuscular and aerosol vaccination. The alternative routes of vaccination were well tolerated and many of the volunteers who received aerosol vaccinations exhibited serum antibody titers that were comparable to those induced by intramuscular vaccination. A mucosal immune response was induced by aerosol vaccination as demonstrated by the induction of anti-HPV16 VLP IgA secreting cells in PBMC and SIgA in secretions. Our data suggest that aerosol administration of HPV VLPs may represent a potential alternative to parenteral injection. PMID- 15882524 TI - Leishmanization: use of an old method for evaluation of candidate vaccines against leishmaniasis. AB - To establish the safety and reproducibility of live challenge with Leishmania major, a single frozen stabilate was used in two open, single-arm leishmanization (LZ) trials. A total of 42 inoculations in 28 male adult volunteers were followed until complete healing. Lesions induced by LZ are as diverse as natural infection, but much milder. Total protection was seen in 100% (11/11) of recovered volunteers. LZ used as live challenge to test vaccine candidates reduces the required sample size, is cost effective for surrogate markers studies and will induce protection in the participants who are not protected by candidate vaccines. PMID- 15882525 TI - A pre-S/S CHO-derived hepatitis B virus vaccine protects woodchucks from WHV productive infection. AB - We evaluated whether a non-adjuvanted vaccine derived from Chinese hamster ovary cells was capable of providing protection against woodchuck hepatitis virus (WHV). Three woodchucks were vaccinated with four 50-microg doses and challenged with a previously characterized virus isolate (WHV197). In all three animals, titre levels of antibodies against hepatitis B surface antigens (anti-HBs) exceeded 10 mIU/ml, peaking at 150 mIU/ml. Challenge resulted in productive acute infection in the two non-vaccinated woodchucks yet in none of the vaccinated woodchucks. In the vaccinated animals, there was evidence of abortive infection. The results demonstrate that a human vaccine is able to protect woodchucks from WHV infection. PMID- 15882526 TI - Combined reduced-antigen-content diphtheria-tetanus-acellular pertussis and polio vaccine (dTpa-IPV) for booster vaccination of adults. AB - Many countries recommend diphtheria, tetanus and/or poliomyelitis boosters in adolescents or adults and the need for pertussis booster vaccination beyond childhood is increasingly recognized. A new combined reduced-antigen-content dTpa IPV vaccine provides booster vaccination against all four diseases in one single injection. The immunogenicity and safety of the dTpa-IPV vaccine was compared to that of licensed dTpa+IPV or Td-IPV vaccines in 806 adolescents >14 years of age and adults with a heterogeneous vaccination history. The dTpa-IPV vaccine was immunogenic and well tolerated. No clinically significant differences were observed between groups. Anti-tetanus antibody kinetics indicated that each of the vaccines could be used for tetanus prophylaxis in acute wound management. For all vaccines, the lowest post-vaccination antibody concentrations were observed in subjects >40 years of age, those seronegative prior to vaccination and those subjects whose last vaccination was > or =20 years ago. In conclusion, dTpa-IPV vaccination of subjects over 14 years of age was as immunogenic and well tolerated as the licensed dTpa+IPV or Td-IPV vaccines. Vaccination coverage of adults is poor and the use of combined vaccines such as dTpa-IPV during vaccination visits, or for wound management, maximizes opportunities for boosting in these difficult to reach age groups. PMID- 15882527 TI - Targeting lymphocyte Peyer's patch adhesion molecule-1: a relay approach to gut immunization. AB - Targeting vaccines to dendritic cells (DCs) can enhance responses to weak vaccine antigens. Although there are molecules that are relatively specific for the various DC subsets, there are none that are both region-specific and DC-specific. This has provided some limitation to targeting regional DC populations. We proposed that these limits could be overcome by targeting antigens not to the DC subsets directly but to cells that persistently seek out and closely interact with DCs, namely lymphocytes. To investigate this hypothesis, we targeted antigens to a unique population of gut-homing lymphocytes and then looked at the induction of immune responses at this site. Using an anti-LPAM-1 (Lymphocyte Peyer's patch adhesion molecule-1; alpha(4)beta(7) integrin) monoclonal antibody (mAb) as a model antigen, we found that targeting gut-homing lymphocytes could significantly elevate the gut mucosal IgA response. Moreover, such a strategy greatly elevated the systemic IgG as well as IgA response. We found that LPAM-1 targeting enhanced the localization of antigen to both the systemic and mucosal lymphoid compartments where both IgA and IgG responses were induced. We also found that any parenteral route of delivery sufficed. Overall, targeting unique populations of lymphocytes may provide a strategy for ferrying antigen to sites that such lymphocytes home to. PMID- 15882528 TI - Influenza vaccination coverage and related factors among Spanish patients with chronic obstructive pulmonary disease. AB - This study sought: to describe influenza vaccination coverages among COPD patients treated in a primary-care setting; and to analyse the factors linked to compliance with vaccination recommendations. This was a descriptive study in a primary-care (PC) setting. Each of the 2422 randomly selected medical practitioners included in the study was required to recruit five COPD patients. Information was drawn from patients' clinical histories and personal interviews. As the dependent variable, we took the answer (yes or no) to the question, "did you have an influenza vaccination in the most recent campaign?"; and as independent variables, we analysed socio-demographic data, health-status related variables, lifestyles and history of pneumococcal vaccination. A total of 10,711 patients were enrolled 87.2% reported having been vaccinated in the most recent campaign. In conclusion, Spanish COPD patients treated in a primary-care setting can be said to enjoy good vaccine coverages against the influenza virus. More frequent contact with the general practitioner and a history of pneumococcal vaccination increase the likelihood of being vaccinated considerably, and measures should be implemented with the aim of improving coverages among younger subjects and those who lead less healthy lifestyles. PMID- 15882529 TI - Multicomponent Lyme vaccine: three is not a crowd. AB - Lyme disease is caused by the spirochete Borrelia burgdorferi and it is the most common vector-borne disease in the United States. Disseminated spirochetes can persist in various tissues and can result in a variety of different disease manifestations. Vaccination trials testing various lipoprotein candidates have yielded mixed results despite the generation of robust antibody titers. Data presented in this report demonstrate that a combination vaccine composed of DbpA, BBK32 and OspC is more effective than single or double component formulations and that the ratio of each component dramatically impacts vaccine efficacy when tested in protection experiments against Borrelia following needle inoculation. PMID- 15882530 TI - Safety of varying dosages of 7-valent pneumococcal protein conjugate vaccine in seniors previously vaccinated with 23-valent pneumococcal polysaccharide vaccine. AB - In a phase I/II dose escalation study, varying volumes (0.1 ml, 0.5 ml, 1.0 ml and 2.0 ml) of 7-valent pneumococcal conjugate vaccine (PCV) (Prevnar or 0.5 ml of 23-valent pneumococcal polysaccharide vaccine (PPV) were administered to 220 adults 70 through 79 years of age previously vaccinated with 0.5 ml PPV at age 65 years or above and at least 5 years previously. Fever was uncommon and did not vary by study group. The rate of local reactions increased with higher volumes of PCV and the rate following 2.0 ml of PCV was comparable to that following 0.5 ml PPV. PMID- 15882531 TI - Comparative study on adjuvanticity of compound Chinese herbal medicinal ingredients. AB - In order to compare the adjuvant activity of compound Chinese herbal medicinal ingredients (cCHMIs) and filtrate best prescription and dose, two initial screened cCHMIs were prepared and mixed with Newcastle disease (ND) vaccine respectively at three doses to vaccinate 17-day-old chickens, taking the non adjuvant and oil adjuvant vaccines as controls. On day 7, 14, 21, 28, 35 and 42 after vaccination, the dynamic changes of peripheral lymphocyte proliferation and serum antibody titers in the chickens were determined by means of MTT method and hemagglutination inhibition test. The results showed that two cCHMIs could promote peripheral lymphocyte proliferation and enhance serum antibody titer, of which adjuvant activity was similar to and even stronger than oil adjuvant in promoting lymphocyte proliferation at some time points, and there was a certain dose-effect relationship. The best adjuvant activity of cCHMIs 2 in promoting cellular immunity occurred at middle dose, and cCHMIs 1 in enhancing humoral immunity, at high dose. Based on good synergistic effects of their components, two cCHMIs would be expected as new-type immunologic adjuvants to substitute for oil adjuvant. PMID- 15882532 TI - In vitro pyrogenicity of the diphtheria, tetanus and acellular pertussis components of a trivalent vaccine. AB - We have earlier found that a trivalent vaccine, containing antigenic components from both Gram-positive and Gram-negative bacteria, induced secretion of the endogenous pyrogen interleukin 6 (IL-6) when added to fresh human blood in vitro. The results of the present study showed that the IL-6 secretion was induced by toxoids derived from the Gram-positive bacterium Corynebacterium diphtheriae. However, fresh whole blood from different donors reacted differently to the stimulation. The blood from some donors induced secretion of large concentrations of IL-6, while the blood from other donors induced essentially no IL-6 secretion as a response to stimulation with diphtheria toxoid or a mixture of diphtheria and tetanus toxoids. Repeated testing over several years using blood from the same donor confirmed a donor-dependency of the reaction. This donor-dependency was only found for the toxoid, since blood from all donors reacted with approximately similar IL-6 production to stimulation by endotoxin from the Gram negative bacterium Escherichia coli, known to be mediated via the toll-like receptor (TLR) 4. Also, no donor-dependecy was found to highly purified lipoteichoic acid from the Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus, known to be mediated via TLR-2 and TLR-6. The receptors involved in stimulation by diphtheria toxoid are not known, but may differ from those used by endotoxin and lipoteichoic acid. PMID- 15882533 TI - Protective vaccination against experimental canine visceral leishmaniasis using a combination of DNA and protein immunization with cysteine proteinases type I and II of L. infantum. AB - Leishmania infantum is known to be associated with visceral leishmaniasis in Iran and canids are natural reservoirs. Control of disease in dogs appears to be one of the most effective approaches for interrupting the domestic cycle of the disease. In search for successful vaccine strategies, we evaluated the cysteine proteinases (CPs) type I and II using a heterologous prime-boost regime for vaccination against experimental visceral leishmaniasis in dogs. Following vaccination and challenge, dogs were followed for 12 months. Ten dogs vaccinated by prime/boost with DNA/recombinant CPs (in combination with CpG ODN and Montanide 720) remained free of infection in their bone morrow. In contrast, three out of four dogs in the control groups had infection in their bone marrow. The peripheral lymphocytes from protected animals had generally higher proliferation responses to F/T antigen, recombinant CPA (rCPA) and recombinant CPB (rCPB) than controls. During post-challenge period, the difference in stimulation index is significant (p<0.05) on months 11 and 12 to F/T antigens, all months for rCPA and 5, 7, 9, 11 and 12 months for rCPB. Analysis of cytokine mRNA level suggested that vaccinated dogs had elevated IFN-gamma mRNA in peripheral blood mononuclear cells (PBMC), whereas there was a consistent increase in the level of IL-10 in the control groups and some vaccinated dogs. The level of total IgG and IgG2, but not IgG1, to rCPA and rCPB was significantly higher in the vaccinated group (p<0.05) than the control groups. We also showed that with the exception of one dog, all dogs in the vaccinated group in comparison to control dogs had strong DTH responses. We propose that the combination of DNA and recombinant protein vaccination using CPs could be instrumental to control (VL) in dogs. PMID- 15882534 TI - Oculo-respiratory syndrome after influenza vaccination: trends over four influenza seasons. AB - BACKGROUND: Oculo-respiratory syndrome (ORS) following influenza vaccination was identified in Canada in 2000. This report describes trends of ORS reported during four consecutive seasons 2000, 2001, 2002 and 2003 in the province of Quebec, Canada. METHODS: Data come from the vaccine-associated adverse event (VAAE) passive reporting system of the Province of Quebec. RESULTS: The rate of ORS reported per 100000 doses distributed declined from 46.6 in 2000 to 34.2, 20.6 and 9 in 2001, 2002 and 2003, respectively. There was no significant difference in rates for ORS between the two vaccines in use in Canada (Fluviral and Vaxigrip) both in 2001 and 2002. During the 4 years, incidence was highest in people aged 40-59 years and declined in older age groups. The clinical profile of ORS has remained remarkably stable over years. Overall, ocular, respiratory symptoms or facial edema were reported by 58%, 84% and 31% of patients, respectively, and 15% had symptoms including all three symptom categories. ORS lasted more than a week in 8-13% of the cases. CONCLUSION: ORS is an adverse event that occurred with both influenza vaccines used in Canada. Its frequency has declined substantially but is still present after 4 years. It constitutes a clinical entity distinct from anaphylactic allergy. Unlike anaphylaxis, ORS does not constitute an absolute contraindication to further doses. PMID- 15882535 TI - In ovo vaccination with the Eimeria tenella EtMIC2 gene induces protective immunity against coccidiosis. AB - An Eimeria tenella microneme recombinant gene (EtMIC2) and encoded protein were evaluated as potential vaccines against avian coccidiosis. In ovo inoculation with the EtMIC2 gene increased anti-EtMIC2 antibody titers at days 10 and 17 following E. tenella infection. In addition, vaccinated birds developed protective immunity against infection by E. tenella as assessed by significantly increased body weight gain and decreased fecal oocyst shedding compared with non vaccinated controls. Vaccination with the EtMIC2 gene also led to protective immunity against infection by E. acervulina, but not E. maxima. Combined in ovo DNA vaccination plus post-hatch boosting with EtMIC2 DNA or protein did not improve antibody titers or protective immunity beyond that achieved with in ovo vaccination alone. These results provide evidence that in ovo immunization with a recombinant Eimeria microneme gene stimulates protective intestinal immunity against coccidiosis. PMID- 15882536 TI - A DNA vaccine expressing the E2 protein of classical swine fever virus elicits T cell responses that can prime for rapid antibody production and confer total protection upon viral challenge. AB - Immunization of domestic pigs with a DNA vaccine expressing the complete E2 protein of classical swine fever virus (CSFV) conferred total protection against a severe viral challenge. Immunization with three doses of plasmid pcDNA3.1/E2 elicited a consistent and specific, MHC class II restricted T cell response in the three domestic pigs analyzed, in the absence of detectable anti-CSFV antibodies in serum. Upon challenge specific T cell responses were boosted in the three vaccinated pigs, and a rapid rise in the titers of CSFV neutralizing antibodies was noticed in two of them, which correlated with a total protection. In these two pigs, neither disease symptoms were observed nor was virus detected at any time after CSFV infection. Neutralizing antibody titers were lower in the third vaccine, which developed a mild and transient peak of pyrexia. As expected, similar analyses in three control pigs (injected with the empty vector or PBS) did not reveal the induction of specific T cells or viral antibodies and, upon challenge, animals developed severe symptoms of the disease, including high titers of viremia, hyperthermia and virus spread to different organs. Control pigs developed, also, a marked leucopenia, resulting in SWC3+ (myelomonocytic cells) being the major PBMC population, and a drastic decrease CD3+ T cells. This T cell depletion was prevented in animals immunized with pcDNA3.1/E2. The total protection achieved, in the absence of CSFV antibodies before challenge, supports the relevance in the antiviral response observed of specific T cell responses primed by pcDNA3.1/E2 vaccine, which, upon challenge, led to a rapid induction of neutralizing antibodies. The observation that CSFV antibodies could only be detected in protected animals after viral challenge opens the possibility of exploring the potential of the DNA vaccine approach used to develop marker vaccines against CSF. PMID- 15882537 TI - Effect of telmisartan/hydrochlorothiazide combination versus nifedipine GITS on ambulatory blood pressure and sympathetic activation. AB - BACKGROUND: This study compares the effects of telmisartan hydrochlorothiazide (HCTZ) combination versus nifedipine GITS on ambulatory blood pressure (BP) and sympathetic activity, in patients with mild-to-moderate hypertension. METHODS: One hundred twenty-four outpatients with sitting diastolic BP > or =95 mmHg and <110 mm Hg were randomized to telmisartan 80 mg/HCTZ 12.5 mg daily (n = 62) or nifedipine GITS 60 mg daily (n = 62) for 12 weeks, according to a prospective, open-label, blind end point, parallel-group design. At the end of a 2-week washout period and after 12 weeks of active treatment, 24-h noninvasive ambulatory BP monitoring (ABPM) was performed, clinic BP and heart rate were measured, and plasma norepinephrine and cardiovascular responses to mental stress induced by the color word test were assessed. RESULTS: Both treatments reduced ambulatory and clinic BP. However, the drug combination had an antihypertensive efficacy significantly greater than nifedipine GITS, as shown by the 24-h (P < .001), daytime (P < .001), and night-time (P < .01) ambulatory BP monitoring values, as well as by the clinic BP at trough (P < .05). The trough-to-peak ratio was similar, but the smoothness index was significantly higher with the combination for both systolic and diastolic BP (P < .05). A significant increase in plasma norepinephrine levels in resting conditions was observed with nifedipine GITS (+20%) but not with telmisartan/HCTZ combination. The color word test produced a greater increase in plasma norepinephrine and heart rate values in the patients treated with nifedipine GITS than in those treated with the combination. CONCLUSIONS: These results suggest that the telmisartan 80 mg/HCTZ 12.5 mg combination provided a more sustained and homogeneous BP control than nifedipine GITS 60 mg, without producing sympathetic activation. PMID- 15882538 TI - White-coat hypertension and autonomic nervous system dysregulation. AB - BACKGROUND: White-coat hypertension, defined as high blood pressure (BP) on clinical assessment but normal BP elsewhere or on ambulatory measurement, is a common but poorly understood phenomenon. The current study asks whether individuals with white-coat hypertension have abnormal autonomic-cardiac regulation, similar to that observed in sustained or persistent hypertension. METHODS: Participants were men (ages 40 to 70 years; 63% white, 37% African American) not receiving any cardiovascular medications who were classified as persistent hypertensive (n = 40), white-coat hypertensive (n = 40), or normotensive (n = 40) on the basis of clinic and daytime ambulatory BP, using a threshold criterion for hypertension of 140/90 mmHg. Persistent and white-coat hypertensive subjects were matched on ethnicity and clinic BP, and white-coat hypertensive subjects and normotensive subjects were matched on race and daytime ambulatory BP. Frequency domain analysis of resting beat-to-beat heart rate variability (HRV) was used to estimate parasympathetic and sympathetic control of the heart. RESULTS: Relative to normotensive subjects, both persistent and white coat hypertensive subjects had lower high-frequency (HF) (P < .03) and low frequency (LF) power (P < .051) and thus less parasympathetic activity. In addition, white-coat and persistent hypertensive subjects had significantly greater LF/HF ratios, indicating greater sympathetic-to-parasympathetic activity, as compared with normotensive subjects (P < .03). CONCLUSIONS: These findings suggest similarities between persistent and white-coat hypertensive subjects reflecting attenuated parasympathetic control of the heart. In addition, the association between white-coat hypertension and autonomic dysregulation, particularly diminished parasympathetic tone, may serve as a mechanism for increased risk for cardiovascular events in affected individuals. PMID- 15882539 TI - In clinical practice, masked hypertension is as common as isolated clinic hypertension: predominance of younger men. AB - BACKGROUND: The prevalence and implications of masked hypertension are under investigation. The aim of this study was to investigate the clinical characteristics associated with masked hypertension in subjects referred for ambulatory blood pressure (BP) monitoring. METHODS: We analyzed 1494 BP monitoring sessions. A subject was considered to have isolated clinic hypertension if clinic BP was > or =140/90 mmHg and awake BP was <135/85 mmHg. Masked hypertension was diagnosed when clinic BP was <140/90 mmHg and awake BP was > or =135/85 mmHg. RESULTS: Of 1494 individuals, 16% had normal BP, 11% had isolated clinic hypertension, 61% had hypertension, and 11% had masked hypertension. Subjects with masked hypertension were younger and more likely to be male than subjects with isolated clinic hypertension, and their awake heart rate was significantly higher. A negative correlation was found between the awake clinic systolic BP difference and clinic systolic BP (r = -0.7, P < .0001). The reproducibility of the masking phenomenon was comparable to that of other variables. CONCLUSIONS: In a group of consecutive subjects referred for ambulatory BP monitoring, masked hypertension was found to be as common as isolated clinic hypertension. Masking was correlated with male sex, young age, and higher awake heart rate, thus suggesting a causal relationship with greater daytime physical activity. The linear association of the masking and the white coat effects to clinic BP suggests that regression toward the mean may partially explain these phenomena. PMID- 15882540 TI - Sleep apnea, hypertension, and the effects of continuous positive airway pressure. AB - BACKGROUND: Sleep apnea is being studied as a risk factor for hypertension. This observational chart review was conducted to determine the long-term effects of continuous positive airway pressure (CPAP) treatment on blood pressure (BP) in a sample of sleep apnea patients from urban and rural populations. METHODS: This study was conducted using data from 180 clinical charts from 1995 to 2002. Patients were identified as hypertensive or normotensive by their initial BP values before they were diagnosed with sleep apnea and were also reviewed after the use of CPAP. RESULTS: Of the patients diagnosed with sleep apnea, 32% were found to have hypertension (mean systolic BP: 164.4 +/- 20.3 mmHg; mean diastolic BP: 96.9 +/- 5.3 mmHg). The average use of CPAP was 12.1 +/- 22.4 months. The hypertensive group showed a significant reduction in BP with CPAP use: systolic BP dropped by an average of 11.2 mmHg (P < .001) and diastolic BP dropped by an average of 5.9 mmHg (P < .001). CONCLUSIONS: Our results confirm that frequency of hypertension is increased among sleep apnea patients. The long-term use of CPAP in hypertensive patients with sleep apnea is associated with a significant decrease in BP to levels that considerably decrease cardiovascular risk. PMID- 15882541 TI - Carotid artery intima-media thickness could predict the presence of coronary artery lesions. AB - The purpose of the present study was to examine whether intima-media thickness (IMT) predicts the presence of the coronary artery lesions independent of other risk factors including clinic blood pressure (BP), parameters of 24-h ambulatory BP monitoring, body mass index, serum cholesterol, and glucose levels. The study population consisted of 390 consecutive subjects who had recently undergone coronary arteriography; 51 subjects with no measurable lesions in their coronary arteries (control group) and 339 subjects with coronary artery lesions (coronary artery disease [CAD] group). Mean IMT of the common carotid artery (MCCA) and internal carotid artery (MICA) were significantly higher in subjects with CAD compare control subjects (P < .0001). Carotid IMT could predict the presence of coronary artery lesions independently of clinic or ambulatory BP values, BMI, serum cholesterol, and glucose levels (P < .01). Carotid IMT predicted the presence of significant coronary artery lesions with cutoff values 0.85 and 0.80 for MICA and MCCA, respectively. The IMT could be a clinical useful test for the presence of significant coronary artery lesions. PMID- 15882542 TI - Nondipping status does not attenuate the conjugated estrogen-induced improvement in aortic stiffness in postmenopausal women with untreated hypertension. AB - BACKGROUND: Whether the blunted nocturnal blood pressure (BP) fall alters the potential beneficial effects of estrogen replacement therapy (ERT) on arterial elastic properties in hypertensive postmenopausal women has not been clarified. The aim of this study was to determine the potential beneficial effects of ERT on arterial elastic properties and to investigate whether a blunted nocturnal BP fall could unfavorably modify the estrogen-induced alterations of large-artery stiffness in postmenopausal women with untreated essential hypertension. METHODS: A total of 66 postmenopausal women with untreated essential hypertension underwent carotid-femoral pulse wave velocity (PWV) measurements at baseline and after 12 weeks of ERT with 0.625 mg conjugated estrogen. By 24-h ambulatory BP monitoring, women were classified according to dipping status (nondippers, n = 21; dippers, n = 45). RESULTS: The two groups were similar regarding age, body mass index, time since menopause, and lipidemic profile. Initially, nondippers compared to dippers, although they had significantly greater office systolic BP (SBP), 24-h SBP, daytime SBP and night-time SBP (by 5, 2, 3, and 19 mm Hg, respectively, P < .05 for all cases), did not differ regarding left ventricular (LV) mass index and aortic PWV (116 v 114 g/m2 and 8.40 v 7.95 cm/sec, respectively, P = NS for both cases). Use of ERT, without affecting the office SBP and DBP, reduced significantly the aortic PWV in both nondippers and dippers (by 1.28 and 1.50 cm/sec, respectively, P < .05 for both cases). However, these PWV reductions were not different between the two groups (P = NS). A multivariate analysis identified patient age and 24-h SBP as significant determinants of estrogen-induced aortic PWV reduction (P < .05). CONCLUSIONS: A blunted nocturnal BP fall does not attenuate the estrogen-induced favorable modifications of large artery elastic properties in hypertensive postmenopausal women. Whether these findings suggest that hypertensive women with ERT-induced attenuation of PWV represent a specific clinical subgroup of patients with possible ERT-associated cardiovascular benefit remains to be determined by properly randomized trials. PMID- 15882543 TI - Relationship between mercury in blood and 24-h ambulatory blood pressure in Greenlanders and Danes. AB - BACKGROUND: Intake of mercury with food items from sea mammals and fish has been suggested to be involved in cardiovascular disease, but the relationship between mercury in blood and 24-h ambulatory blood pressure (BP) has never been studied. METHODS: We measured mercury in blood and 24-h BP in four groups of healthy subjects: group 1, Danes living in Denmark consuming European food; group 2, Greenlanders living in Denmark consuming European food; group 3, Greenlanders living in Greenland consuming European food; and group 4, Greenlanders living in Greenland consuming mainly traditional Greenlandic food. RESULTS: Mercury in blood was highest in Greenlanders and increased when they lived in Greenland and consumed traditional Greenlandic food (group 1: 2.2 microg/L (median), group 2: 4.8 microg/L, group 3: 10.8 microg/L, and group 4: 24.9 microg/L). The 24-h BP was the same in all three groups of Greenlanders. However, 24-h diastolic BP was lower among Greenlanders than Danes (71 v 76 mm Hg, P < .000) and 24-h pulse pressure was higher (54 v 50 mm Hg, P < .000). Mercury in blood was significantly and positively correlated to pulse pressure (rho = 0.272, P < .01). CONCLUSIONS: Pulse pressure was higher and diastolic BP was lower in Greenlanders than Danes. Pulse pressure increased with higher mercury content in the blood. Although genetic factors must be responsible to some extent for the difference in pulse pressure between Greenlanders and Danes, the present results seem to support the hypothesis that mercury intake from maritime food is involved in cardiovascular disease. PMID- 15882544 TI - Resistant hypertension revisited: a comparison of two university-based cohorts. AB - BACKGROUND: More than a decade ago, we found that a suboptimal medication regimen was the leading cause of resistant hypertension (RH) among patients referred to a tertiary care clinic. Since then, lower blood pressure (BP) goals have been recommended, suggesting that more patients may have RH. To assess whether the reasons for and treatment of RH have changed, we determined the frequency of various causes of resistance, the proportion of patients achieving goal BP, and the changes made in antihypertensive regimens. METHODS: The charts of all new patients seen at the RUSH University Hypertension Center between January 1, 1993, and November 1, 2001, were reviewed for strict criteria for RH: 1) physician referral for uncontrolled hypertension; 2) BP > or =140/90 mmHg despite use of three antihypertensive drugs; and 3) at least one follow-up visit. Patients were followed-up until goal BP was achieved on two consecutive visits or their last visit or until March 2002. RESULTS: Of 1281 patients, 141 met criteria for RH. A cause of resistance was found in 94% of cases, including the following: drug related causes (58%); nonadherence (16%); psychological causes (9%); office resistance (ie, in-clinic BP readings that were higher than goal despite treatment with antihypertensive medications and despite normotensive BP outside of the clinic as demonstrated by 24-h ambulatory BP monitoring) (6%); and secondary hypertension (5%). Overall, 53% of patients had their BP controlled to <140/90 mmHg, largely from regimen optimization and intensification, proper use of diuretics, and on average 4.1 +/- 1 antihypertensive medications (3.7 +/- 0.9 on referral). CONCLUSIONS: These data are strikingly similar to those from our previous study of RH, in which a suboptimal medication regimen was the most common reason for resistance. Goal BP was most commonly achieved after optimizing the diuretic regimen and increasing the number of medications, suggesting that physicians should use these measures to attain the recommended lower BP goals If goal BP is not reached, referral to a clinical hypertension specialist may be appropriate. PMID- 15882545 TI - A genome scan for linkage with aortic root diameter in hypertensive African Americans and whites in the Hypertension Genetic Epidemiology Network (HyperGEN) study. AB - BACKGROUND: Aortic root dilation is a prominent feature in several cardiovascular diseases. This study seeks to identify genomic regions linked to variation in the aortic root diameter (ARD) in hypertensive African American and white individuals. METHODS: We performed a genome scan for ARD in the Hypertension Genetic Epidemiology Network Study, one of four networks in the National Heart, Lung, and Blood Institute Family Blood Pressure Program (FBPP). Data were collected from 1129 African American siblings from 504 hypertensive sibships and 883 white siblings from 374 hypertensive sibships. Standardized residual values of ARD were calculated using linear regression, adjusting for effects of age, age2, and field center (ie, minimally adjusted model), separately in groups composed by sex and ethnicity. The ARD was additionally adjusted for height, weight, diastolic BP, and systolic BP in a fully adjusted model. Multipoint linkage analysis was performed using the GENEHUNTER2 variance components method. RESULTS: Suggestive evidence for linkage was found on chromosome 5 at 85 cM in African Americans, with a maximal log of the odds (LOD) score of 2.07. Suggestive evidence for linkage was found on chromosome 1 at 157 cM in whites, with a maximal LOD score of 2.40. CONCLUSIONS: Our findings suggest that genes present on chromosomes 1 and 5 might influence inter-individual variation in aortic root diameter. PMID- 15882546 TI - Common genetic mechanisms of blood pressure elevation in two independent rodent models of human essential hypertension. AB - Genetic studies of essential hypertension, a complex, polygenic, and age dependent disorder, have not been able to completely elucidate the genes responsible for development of the trait. We used a novel strategy to compare gene expression in the adrenal gland of two independent rodent models of human essential hypertension (the spontaneously hypertensive rat, SHR, and the blood pressure high mouse, BPH), with the goal of uncovering shared, common genetic mechanisms of hypertension across mammalian species that might, therefore, be pertinent to human hypertension. We deliberately studied young, 4- to 5-week-old, "prehypertensive" SHR and BPH that had not yet developed complete elevations in blood pressure (BP), so that we could minimize the impact of chronic, sustained BP elevation, age, and other confounding factors on gene expression, therefore increasing the likelihood that differential expression reflects relatively early pathogenic mechanisms in hypertension, rather than later responses to, or compensations for BP elevation. We compared transcript expression patterns of genes orthologous between the rat and the mouse, and presented candidate genes for hypertension that are differentially expressed in the same direction in SHR and BPH (ie, overexpressed in both SHR and BPH, or underexpressed in both SHR and BPH). Then we used a systems biology approach to analyze expression patterns in biochemical pathways and networks to isolate systems involved in hypertension pathology in both SHR and BPH. We found transcript pattern evidence for involvement of several systems in the pathology of hypertension in SHR and BPH: adrenal catecholamines and sympathetic function; steroid hormone synthesis, catabolism, and its contribution to enhanced glucocorticoid sensitivity in SHR; oxidative stress and its role as a common mechanism of vascular and end-organ injury; and intermediary metabolism with global but mechanistically different perturbations in SHR and BPH. Approximately 10% of the differentially expressed orthologous genes we studied shared a common direction of expression in the two hypertensive rodent strains, suggesting fundamental transcriptional mechanisms in common whereby mammals can elevate BP or respond to such elevation; even these shared orthologs spanned a diverse set of biological processes, reinforcing the multifactorial and complex nature of hypertension. PMID- 15882547 TI - Genome-wide linkage study of erythrocyte sodium-lithium countertransport. AB - BACKGROUND: Increased erythrocyte sodium-lithium countertransport (SLC) has been observed in patients with essential hypertension. An analytic strategy for identification of genetic variation contributing to hypertension is the evaluation of appropriate intermediate phenotypes for hypertension, such as SLC. Thus, in this study, genome-wide linkage scans for SLC were performed in two independent samples of pedigrees from the Rochester Family Heart Study (RFHS). METHODS: Genome-wide linkage scans for SLC were performed in independent samples of 232 and 252 non-Hispanic white families from the RFHS. Multipoint variance component linkage analysis was performed using MERLIN. RESULTS: Chromosomes 8, 9, 10, 19, and 20 contained evidence for linkage (log of the odds [LOD] > or =2) in at least one sample of RFHS pedigrees. Consistent evidence of linkage for SLC between the two samples was observed on chromosome 10 (LOD = 2.02 at 64 cM in the first sample and LOD = 2.27 at 55 cM in the second sample). CONCLUSIONS: Consistent evidence of linkage (LOD > or =2) for SLC was observed in two independent samples of non-Hispanic white families on chromosome 10. Concordance of linkage evidence between the two samples provides confidence that a region on chromosome 10 contains genetic variation influencing SLC, which may potentially influence susceptibility to hypertension. PMID- 15882548 TI - Clinical and genetic correlates of serum aldosterone in the community: the Framingham Heart Study. AB - BACKGROUND: We investigated the environmental and genetic sources of interindividual variability in serum aldosterone level in a large, community based sample. METHODS: We examined the relation of serum aldosterone to vascular risk factors, urine sodium, and candidate single nucleotide polymorphisms in 2891 Framingham Offspring Study participants (53.2% women, mean age 59 years) using multivariable linear regression. Multivariable logistic regression was used to identify predictors of high (top quartile) and low (lowest quartile) serum aldosterone values. We estimated heritability of serum aldosterone via variance component methods and evaluated linkage via a 10-cM-density genome scan. RESULTS: Clinical variables related to higher serum aldosterone level included female sex, diuretic treatment, and a higher total/high density lipoprotein cholesterol ratio. A high urinary sodium excretion, postmenopausal status (without hormone replacement therapy), increased pulse pressure, and prevalent cardiovascular disease were related to lower serum aldosterone values. Urinary sodium was the strongest correlate of serum aldosterone (R2= 10%). Serum aldosterone levels did not differ by genotype in the aldosterone synthase (CYP11B2c.1-344C>T) and the mineralocorticoid receptor (NR3C2c.754A>G) genes. The estimated heritability of serum aldosterone was 0.10. No chromosomal region attained a log-of-the-odds score >1 in multipoint linkage analysis. CONCLUSIONS: We observed a complex relation between serum aldosterone and vascular risk factors. The genetic contribution to serum aldosterone level was modest. PMID- 15882549 TI - Quantitative trait loci mapping for intracellular calcium in spontaneously hypertensive rats. AB - BACKGROUND: Increased intracellular calcium ([Ca2+]i) in platelets is also proposed as an intermediate phenotype for hypertension in spontaneously hypertensive rats (SHR). Increased [Ca2+]i in platelets is hypothesized to contribute to atherothrombotic events. Platelet hyperactivity is frequently associated with cardiovascular disease. METHODS: In a genome scan, we performed the quantitative trait loci (QTL) mapping for [Ca2+]i in back-crossed rats derived from SHR and normotensive Fischer 344 rats, which demonstrated a single major QTL for hypertension on chromosome 1. Thrombin-stimulated [Ca2+]i in Ca2+ free and in Ca2+-containing buffers was measured in platelets using the Fura-2 method. RESULTS: Among the parental strains, systolic blood pressure and thrombin stimulated [Ca2+]i were significantly greater in SHR than in Fischer 344 and F1 rats. The sarco(endo)plasmic reticulum Ca2+-dependent ATPase II gene locus (Serca2) between D12Mgh5 and D12Mgh6 showed the significant linkage for thrombin stimulated [Ca2+]i in Ca2+-free and Ca2+-containing buffers. The peak logarithm of the odds scores were 3.6 and 3.3, respectively. These QTL explained 19.8% and 17.4% of the total variances, respectively. D3Mit13 and DXMgh1 showed suggestive linkage for thrombin-stimulated [Ca2+]i in Ca2+-free and in Ca2+-containing buffers, respectively. The peak logarithm of the odds scores were 2.6 and 2.1, respectively. CONCLUSIONS: A significant QTL for [Ca2+]i was mapped near Serca2 on chromosome 12, and suggestive QTL were identified near D3Mit13 and DXMgh1 in a genome scan. Genetic abnormalites in platelet [Ca2+]i may contribute to cardiovascular disease via platelet hyperactivity, independent of blood pressure elevation. PMID- 15882550 TI - Further evidence of a quantitative trait locus on chromosome 18 influencing postural change in systolic blood pressure: the Hypertension Genetic Epidemiology Network (HyperGEN) Study. AB - BACKGROUND: Postural change in systolic blood pressure (SBP) is prospectively associated with several disease outcomes including hypertension, stroke, and coronary heart disease. The objective of this study was to characterize further a possible quantitative trait locus on chromosome 18q21 influencing SBP response to a postural challenge. METHODS: A prior genome scan of postural SBP response in 636 subjects of white ethnicity from 285 hypertensive sibships in the Hypertension Genetic Epidemiology Network (HyperGEN) indicated suggestive evidence for linkage on chromosome 18q21. This study included a de novo set of 452 African American pedigrees from the HyperGEN study and an expanded set of 372 white pedigrees. Variance components linkage analysis of postural SBP change was conducted using microsatellite markers on chromosome 18, and association studies were performed with a common single nucleotide polymorphism (variant 13) in the gene encoding NEDD4L, a key regulator of fine sodium reabsorption in the distal nephron. RESULTS: Combined analysis of all white and African American pedigrees yielded a LOD score of 4.25 at 80 cM on chromosome 18q21, with at least nominal evidence of linkage at this position in both white (LOD: 3.43) and African American (LOD: 1.14) subjects. Postural SBP response was associated with variant 13 of the NEDD4L in a subset of white subjects taking medications effective in treating sodium volume-dependent hypertension (alpha1-blockers, calcium channel blockers, and/or diuretics). CONCLUSION: These data provide further evidence for a quantitative trait locus on chromosome 18q21 influencing postural change in SBP. PMID- 15882551 TI - Augmentation index and systolic load are lower in competitive endurance athletes. AB - BACKGROUND: Arterial stiffness increases with age, and chronic endurance exercise has been shown to attenuate increases in arterial stiffness in older individuals. The aim of this study was to assess the effect of varying fitness levels on wave reflection in young healthy adults. METHODS: A total of 32 subjects (16 competitive endurance athletes and 16 recreationally active subjects) underwent brachial artery blood pressure (BP) measurement (by sphygmomanometry) and central aortic pressure (by noninvasive radial artery applanation tonometry and use of a generalized transfer function) measurements at rest. Central aortic augmentation index (AI), an index of wave reflection, and tension-time index (TTI), an indicator of systolic load, were calculated from the aortic pressure waveform. Physical fitness was determined through a maximal oxygen consumption (VO2max) test performed on a treadmill. RESULTS: The two groups did not differ in age, height, weight, resting BP, or blood lipids. The VO2max was significantly higher in the competitive group compared with the recreational group (65 +/- 1.9 v 49 +/ 1.8 mL/kg/min, P < .05). The AI was lower in the competitive group compared with the recreational group (-2.1% +/- 2.1% v 4.5% +/- 2.9%, P < .05), as was TTI (1679 +/- 61 v 1868 +/- 58, P < .05). CONCLUSIONS: Wave reflection is lower in competitive endurance athletes who have higher fitness levels and who exercise at a higher intensity, for a longer duration, and more frequently as compared with recreationally active individuals. These differences may be due to functional changes that occur as a result of training. PMID- 15882552 TI - Distribution and correlates of arterial compliance measures in asymptomatic young adults: the Bogalusa Heart Study. AB - BACKGROUND: Impaired arterial compliance is an independent predictor of early vascular damage and related adverse cardiovascular (CV) outcome. Information is scant on the distributions and correlates of measures of arterial pulsatile function in a community-based, biracial cohort of young adults. METHODS: In 800 African American and white subjects aged 18 to 44 years, pulsatile arterial function was assessed in terms of large artery (capacitive) compliance, small artery (oscillatory) compliance, systemic vascular resistance, and vascular impedance by noninvasive radial artery pressure pulse contour analysis. RESULTS: African Americans versus whites and women versus men had lower large and small artery compliances and higher systemic vascular resistance and vascular impedance (P < .001). In multiple regression analysis, mean arterial pressure, body mass index (BMI), insulin levels, and age were correlated inversely and body surface area positively with large artery compliance and accounted for 39.2% of the variance; mean arterial pressure, female gender, age, and triglyceride levels inversely and cardiac output positively with small artery compliance and explained 56.4% of the variance; mean arterial pressure and age positively and cardiac output inversely with systemic vascular resistance and accounted for 91.4% of variance; and mean arterial pressure and BMI positively and cardiac output and body surface area inversely with vascular impedance and contributed to 37.6% of the variance. CONCLUSIONS: The observed deleterious impact of traditional CV risk factors on the arterial wall dynamics in asymptomatic young adults has important implications for preventive cardiology. Noninvasive pulsatile arterial function assessment may be helpful for evaluation of early vascular damage in a high-risk young population group. PMID- 15882553 TI - Extracellular matrix remodeling in the heart of the homocysteinemic obese rabbit. AB - Despite the strides made toward understanding cardiac abnormalities in obesity induced hypertension, the composition and concentration of cardiac extracellular matrix (ECM) components resulting from diet-induced obesity are largely unknown. Previous studies from our laboratory have demonstrated differential expression of collagens, growth factors, and homocysteine (Hcy) in pressure overload models of cardiac hypertrophy. The hypothesis of the present study was that left ventricular hypertrophy (LVH) from the combined pressure and volume overload of obesity induced cardiac fibrosis in part by increasing Hcy, increasing transforming growth factor-beta1 (TGF-beta1), and decreasing decorin. Using the rabbit model, we examined the changes in cardiac collagen accumulation, plasma Hcy, left ventricular (LV) TGF-beta1, and LV decorin after 12 weeks of developing obesity. Cardiac fibrosis was analyzed by trichrome stain for collagens. Total collagens types I and III, TGF-beta1, and decorin were analyzed in tissue homogenates by immunoblots and quantitated with a densitometer. After 12 weeks, rabbits eating a high-fat diet had greater body weight (5.38 +/- 0.3 kg v 3.73 +/ 0.6 kg) and greater LV weight (5.08 +/- 0.05 g v 3.86 +/- 0.17 g) compared with lean rabbits. Heart rate was also significantly higher in obese than in lean rabbits (221 +/- 8 v 173 +/- 5 beats/min). Plasma concentrations of circulating Hcy were 16.9 +/- 2.4 micromol/L and 24.3 +/- 1.8 micromol/L in lean and obese rabbits, respectively. Compared with lean rabbits, obese rabbits had increased interstitial and perivascular collagen, a 4-fold increase in the medial/lumen ratio of coronary vessels, a 1.75-fold increase in cardiac collagen I, and a 1.5 fold increase in cardiac collagen III levels. Levels of TGF-beta1 were increased 1.75-fold, whereas decorin levels were significantly reduced in obese compared with lean rabbits. In conclusion, a high-fat diet, even over a period as short as 12 weeks, causes fibrosis in coronary vessels as well as accumulation of collagen in the cardiac interstitium. The accumulation of cardiac collagen was associated with induction of Hcy and TGF-beta1 and with suppression of decorin. PMID- 15882554 TI - Persistent effects on blood pressure and renal function of perindopril alone or combined with losartan in Lyon hypertensive rats. AB - BACKGROUND: In Lyon genetically hypertensive (LH) rats, an early and long-term angiotensin-converting enzyme (ACE) inhibition with perindopril prevents their hypertension and renal alterations. The present work aimed to determine whether: 1) these effects persist after treatment withdrawal; 2) a short-term additional angiotensin II type 1 (AT1) receptor blockade with losartan potentiates these effects; and 3) an early combination of low doses of perindopril and losartan produces the same prolonged effects as monotherapy with a higher dose of perindopril. METHODS: Perindopril (0.4 or 3 mg/kg/day orally) was given from 3 to 12 weeks of age to male LH rats. In other perindopril-treated LH rats, losartan (10 mg/kg/day orally) was added 1 week before perindopril withdrawal and during a 3-week period. In another group of LH rats, perindopril (0.1 mg/kg/day) and losartan (2.5 mg/kg/day) were given together from 3 to 12 weeks. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were monitored using radio telemetry and renal function studied in anesthetized rats. RESULTS: Eight weeks after perindopril withdrawal, a significant reduction (P < .05) in SBP and DBP was observed with the both doses (SBP: 135 +/- 3, 139 +/- 5 v 157 +/- 4; DBP: 89 +/- 4, 93 +/- 5 v 111 +/- 3 mm Hg for 0.4 and 3 mg/kg/day v untreated LH rats). This was accompanied by a persistent decrease in urinary protein excretion and a long-lasting improvement in pressure natriuresis. The persistent antihypertensive effect was not improved by short-term addition of losartan. Interestingly, the early combination of perindopril with losartan at fourfold lower doses produced similar persistent antihypertensive and renoprotective effects. CONCLUSIONS: The blood pressure reduction produced by an early ACE inhibition in LH rats persists long after treatment withdrawal and is associated with an improvement in renal function. The combination of low doses of perindopril and losartan reveals a long term effect similar to that of a monotherapy with a higher dose of perindopril. PMID- 15882555 TI - Tempol attenuates the development of hypertensive renal injury in Dahl salt sensitive rats. AB - BACKGROUND: Dahl salt-sensitive (DS) rats given a high-salt diet develop renal lesions that are virtually identical to those in human hypertensive nephrosclerosis and are associated with increased oxidative stress. This study looks at the effects of a superoxide scavenger in preventing of hypertensive renal damage in high-salt-treated DS rats. METHODS: The DS rats (n = 5 per group) were treated with 0.3% NaCl diets (LS), 8% NaCl diets (HS), and 8% NaCl diets plus 10 mmol/L tempol in drinking water (HS+T) for 5 weeks. Systolic blood pressure (SBP) was measured by the tail-cuff method. As markers of renal damage, we measured serum creatinine, creatinine clearance, histopathologic indices, and transforming growth factor-beta1 (TGF-beta1; a mediator for renal fibrosis) expression. In addition, 8-hydroxy-2'-deoxyguanosine (8-OHdG)-positive cells and expression of heme oxygenase-1 (HO-1) were quantified as markers of oxidative stress. RESULTS: We found that a high-salt diet (8% NaCl) led to the development of hypertension, increased oxidative stress in the renal tissue (8-OHdG immunoreactive staining and HO-1 protein expression), increased renal histopathologic damage (arteriosclerosis index, matrix score, and interstitial volume) accompanied by accumulation of TGF-beta1, and decreased creatinine clearance in the DS rats. These adverse effects of salt were prevented by the tempol supplementation. CONCLUSIONS: Histopathologic and biochemical findings indicate that, in the DS rat, salt-induced hypertensive nephropathy is associated with increased oxidative stress. Superoxide mimetic tempol can reduce this detrimental effect of salt feeding through TGF-beta1 suppression and consequently prevent the development of hypertension and hypertensive nephropathy. PMID- 15882556 TI - Caffeine tolerance is incomplete: persistent blood pressure responses in the ambulatory setting. AB - BACKGROUND: Caffeine in dietary doses is a well-established pressor agent. Tolerance to this pressor effect occurs in only about half of regular consumers in acute laboratory tests. The clinical significance of this incomplete tolerance depends on whether the pressor effect is maintained throughout the day with repeated intake. Therefore, we examined the ability of a standard dose of caffeine (250 mg x 3) to maintain a blood pressure (BP) elevation during 18 hours of ambulatory BP monitoring (ABPM) after 5 days of regular daily intake of varying background doses. METHODS: Eighty-five men and women completed a four week double blind, crossover trial. During each week, subjects consumed capsules totaling 0, 300, or 600 mg/day of caffeine in 3 divided doses. On day 6, they consumed capsules with either 0 or 250 mg at 9:00 am and 1:00 pm, in the laboratory, and again at 6:00 pm during ABPM. Tolerance was defined as a reduction in the diastolic BP response to two challenge doses given in the lab in response to increasing daily intake. Data were analyzed using multivariate repeated measures analysis of variance. RESULTS: BP responses to caffeine above those found on placebo-placebo (P-P) week were found for both tolerance groups when caffeine was consumed after a week of receiving a placebo. However, only the low tolerance group showed increases, above those found on P-P week, after 300 mg/day in systolic/diastolic BP during the waking hours (mean +/- standard error of the mean = 2.8 +/- 1.1, P = .01/2.2 +/- 0.9, P = .02) and in systolic BP during sleep (2.3 +/- 1, P = .03). CONCLUSIONS: Persistent elevations in BP occurring on a daily basis in some habitual caffeine consumers may hold clinical significance. PMID- 15882557 TI - Mechanisms for the clinical benefits of angiotensin II receptor blockers. AB - The renin-angiotensin-aldosterone system (RAAS) regulates sodium balance, fluid volume, and blood pressure (BP). It is also implicated in the progression of heart failure, hypertension, and kidney disease; this is mediated by the binding of angiotensin II (ang II) to the ang II type 1 but not the ang II type 2 receptor. Preclinical and clinical studies have shown that blockade of the RAAS with ang II receptor blockers (ARB) is effective not only in controlling BP but also in preventing end-organ damage. Through their mechanism of action, ARB may offer benefit with respect to endothelial dysfunction and vascular remodeling, as well as cardiac and renal protection. In addition, both recent and ongoing clinical trials help to clarify further the mechanisms for the benefits of using ARB across the cardiovascular continuum. PMID- 15882558 TI - The association of heart failure with insulin resistance and the development of type 2 diabetes. AB - Heart failure (HF) is associated with a significant risk for either concurrently having diabetes or subsequently developing diabetes. Medications that block the sympathetic nervous system (SNS) have been shown to improve clinical outcomes in patients with chronic HF but may be associated with the development of new-onset diabetes. Depending on the receptor specificity of the individual agent, beta blockers have different effects on glucose and lipid metabolism as well as on the risk for developing new-onset diabetes. Although SNS modulation with beta blockade reduces mortality in patients with chronic HF, certain beta-blockers may actually worsen glucose and lipid metabolism and increase the risk for new-onset diabetes. The use of vasodilating beta-blockers, on the other hand, has not produced these harmful metabolic effects. PMID- 15882559 TI - Synovitis: a potential predictive factor of structural progression of medial tibiofemoral knee osteoarthritis -- results of a 1 year longitudinal arthroscopic study in 422 patients. AB - OBJECTIVE: To evaluate the prevalence of synovitis in painful medial tibiofemoral knee osteoarthritis (OA) and to evaluate correlation between synovitis and the structural severity and progression of tibiofemoral cartilage damage. DESIGN: STUDY: Multicenter, longitudinal, 1-year duration. PATIENTS: Primary painful knee OA (ACR criteria) of the medial tibiofemoral compartment, with pain of the signal knee on at least 30 days in the past 2 months, medial joint space width > or = 2mm, at least 10% of one cartilage surface of the medial compartment affected by superficial fibrillation or worse at baseline arthroscopy. ARTHROSCOPIC PARAMETERS: Knee arthroscopy under local anesthesia was performed and videorecorded at entry and after 1 year. Medial chondropathy was scored by using Societe Francaise d'Arthroscopie (SFA) score (0-100) and reader's overall assessment (VAS score, 100 mm). Progression of medial chondropathy was defined by a change in SFA and VAS scores over 4.5 and 8.0 mm after 1 year, respectively. Medial perimeniscal synovium was scored as normal (few translucent and slender villi, fine vascular network), reactive (proliferation of opaque villi), or inflammatory (hypervascularization and/or proliferation of hypertrophic and hyperemic villi). Medial chondropathy and synovitis were scored by a single reader blind to chronology of paired videotapes. RESULTS: Four hundred and twenty two patients were enrolled (mean age: 61 years, females: 59%, body mass index: 31, mean disease duration: 4 years) and completed the 1-year study. Synovial abnormalities were present in 50% of the patients with reactive and inflammatory aspects in 29% and 21% of the patients, respectively. Patients with a reactive or inflammatory medial synovium had a more severe medial chondropathy. The worsening in medial chondropathy after 1 year was statistically more severe in the group of patients with an inflammatory perimeniscal synovial membrane at baseline compared to patients with normal and reactive aspects, with no difference between these two latter groups. The odds ratio for progression in VAS score after 1 year was 3.11 (95% CI [1.07, 5.69]) for patients with inflammatory synovium at baseline compared to patients with normal synovium. CONCLUSIONS: This study suggests that abnormalities of the medial perimeniscal synovium are a common feature of painful medial knee OA, associated with more severe medial chondropathy. It also suggests that an inflammatory aspect of the medial perimeniscal synovium could be considered as a predictive factor of subsequent increased degradation of medial chondropathy. PMID- 15882560 TI - Arthroscopy -- a potential "gold standard" for the diagnosis of the chondropathy of early osteoarthritis. AB - OBJECTIVES: The aims of this study were to: 1. Evaluate the performance of arthroscopy for the diagnosis of chondropathy and to compare it to that of direct non-arthroscopic assessments; 2. Determine intra-observer reliability of arthroscopic assessments; 3. Evaluate the effects of the arthroscopic video quality and probing upon diagnostic performance. DESIGN: The ovine medial meniscectomy (MMx) model of early osteoarthritis (OA) was used assuming that pre MMx articular cartilage (AC) was "normal" and post-MMx AC "chondropathic". Video recordings of arthroscopic assessments of each stifle compartment were evaluated. Scores were given for the quality of the video and the amount of probing. The diagnostic performances of dynamic shear modulus (G), light microscopic assessment and superficial zone collagen birefringence assessments were evaluated and compared to that of arthroscopy. Intra-observer reliability of arthroscopic assessments was also evaluated. RESULTS: Arthroscopic assessments had high sensitivity (91-100%), specificity (62-88%) and accuracy (75-93%) for the diagnosis of chondropathy 16 weeks after MMx. Arthroscopy compared favourably with the direct non-arthroscopic assessments in the lateral compartment and was found to have extremely high intra-observer reliability (kappa 0.78-1.00). The quality of arthroscopic video recordings and the amount of probing did not significantly influence accuracy or reliability. CONCLUSIONS: Arthroscopy performs as well as direct non-arthroscopic assessments of AC for diagnosis of early OA. These results suggest that arthroscopy can be used as a "gold standard" for the validation of non-invasive assessments like magnetic resonance imaging and that arthroscopic diagnosis can be based on small amounts of video footage without AC probing. PMID- 15882561 TI - The effects of minor hip flexion, abduction or adduction and x-ray beam angle on the radiographic joint space width of the hip. AB - OBJECTIVE: Joint pain may cause patients to hold their limbs in mild flexion, abduction or adduction to minimize pain, regardless of the extent of articular pathology, and these positional changes may have substantial effects on the interpretation of radiographic joint space. We aimed to study the impacts of minor degrees of flexion, abduction or adduction of the hip, as well as the angle of the x-ray beam on the radiographic joint space width (JSW) of the hip joint. METHODS: In the first part of the study, 65 patients (44 males, 21 females, mean+/-SD age 49+/-17) without clinical evidence of hip osteoarthritis (OA) who underwent intravenous pyelography (IVP) were studied. The hips were differentially positioned during the sequential radiographs required for the IVP procedure. Baseline radiographs were taken at 15 degrees internal rotation of the hips [the standard position for anteroposterior (AP) pelvis radiography]; additional positions included the hips at 15 degrees and/or 30 degrees flexion, and 15 degrees adduction and/or 30 degrees abduction. Radiographic JSWs were measured at three sites using a dial caliper: superomedial, superolateral, and the point of narrowest JSW. In the second part, 15 patients without clinical evidence of hip OA who underwent supine abdominal radiography for non rheumatological indications were evaluated by standard (AP) pelvis x-ray in the same setting and JSW measurements were made as described above. RESULTS: When the average of the three measurements of the JSW was taken for each hip, baseline JSW was 4.38+/-0.55 mm (mean+/-SD). Positioning of the hip significantly (P<0.01) affected the radiographic JSW, with apparent widening during adduction and 30 degrees flexion, (JSW 4.56+/-0.51 mm and 4.53+/-0.58 mm, mean+/-SD), respectively, but narrowing during abduction (4.17+/-0.59 mm, mean+/-SD). Fifteen degree flexion of the hip did not result in statistically significant change in JSW measurements. Upon comparison of the AP pelvis view with the supine abdominal view, neither the average JSW nor the point of narrowest JSW differed significantly, although the superolateral JSW was significantly greater on the AP pelvis view (P=0.02). CONCLUSIONS: Subtle positional changes in the hip, such as may occur during pain or in OA, may artifactually alter the measured radiographic JSW. Thus, longitudinal studies which employ hip JSW to assess disease progression may yield biased results due to changes in pain rather than structure unless care is taken to ensure constant positioning of the hip. PMID- 15882562 TI - Glucosamine and chondroitin sulfate regulate gene expression and synthesis of nitric oxide and prostaglandin E(2) in articular cartilage explants. AB - OBJECTIVE: Glucosamine (GLN) and chondroitin sulfate (CS) are widely used to alleviate symptoms of osteoarthritis (OA). However, the mechanism(s) of action of these nutraceuticals remains unresolved. In the present study, we determined the effect of physiologically relevant concentrations of GLN and CS on gene expression and synthesis of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in cytokine-stimulated articular cartilage explants. METHODS: Using bovine articular cartilage explants in culture stimulated with IL-1, the effects of physiologically relevant concentrations of GLN and CS on gene expression of inducible nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS), cyclooxygenase-2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGEs1) were assessed with quantitative real-time polymerase chain reaction (Q-RT-PCR). The production of NO and PGE(2) was also quantified. RESULTS: CS and the GLN and CS combination at concentrations attainable in the blood down-regulated IL-1 induced mRNA expression of iNOS at 24 and 48 h post-culture. Up-regulated iNOS expression at 24h by IL-1 was also suppressed by GLN. GLN and CS transiently repressed the cytokine-stimulated mPGEs1 transcript. Synthesis of NO was reduced with CS alone and the combination after 24h of culture. Repression of COX-2 transcripts by GLN and CS was accompanied by concomitant reduction in PGE(2). CONCLUSION: Our results indicate that physiologically relevant concentrations of GLN and CS can regulate gene expression and synthesis of NO and PGE(2), providing a plausible explanation for their purported anti-inflammatory properties. PMID- 15882563 TI - Chondrocyte calcium-sensing receptor expression is up-regulated in early guinea pig knee osteoarthritis and modulates PTHrP, MMP-13, and TIMP-3 expression. AB - OBJECTIVE: Growth plate chondrocytes up-regulate calcium-sensing receptor (CaR) expression as they mature to hypertrophy. In cells other than chondrocytes, extracellular calcium-sensing via the CaR functions partly to promote expression of parathyroid hormone-related protein (PTHrP), a critical regulator of endochondral development. Moreover, PTHrP is up-regulated in human osteoarthritis (OA) and surgically induced rabbit OA cartilages and may promote both chondrocyte proliferation and osteophyte formation therein. Hence, we examined chondrocyte CaR-mediated calcium-sensing in OA pathogenesis. METHODS: We studied spontaneous knee OA in male Hartley guinea pigs. We also evaluated cultured bovine knee chondrocytes and immortalized human articular chondrocytes (CH-8 cells), employing the CaR calcimimetic agonist NPS R-467 or altering physiologic extracellular calcium (1.8 mM). RESULTS: Immunohistochemistry revealed that CaR expression became up-regulated in the superficial zone at 4 months of age in the guinea pig medial tibial plateau cartilage as early OA developed. CaR expression later became up-regulated in the middle zone. PTHrP content, measured by immunoassay, was significantly increased in the medial tibial plateau cartilage as OA developed and progressed. In cultured chondrocytic cells, CaR-mediated extracellular calcium-sensing, stimulated by the calcimimetic NPS R-467, induced PTHrP and matrix metalloproteinase (MMP)-13 expression and suppressed expression of tissue inhibitor of metalloproteinase (TIMP)-3 dose-dependently, effects shared by elevated extracellular calcium (3 mM). Extracellular calcium-sensing appeared essential for PTHrP and interleukin (IL)-1 to induce MMP-13 and for PTHrP 1-34 to suppress TIMP-3 expression. CONCLUSIONS: Chondrocyte CaR expression becomes up-regulated early in the course of spontaneous guinea pig knee OA. Chondrocyte CaR-mediated extracellular calcium-sensing promotes PTHrP expression, modulates the effects of PTHrP and IL-1, and promotes MMP-13 expression and TIMP 3 depletion. Our results implicate up-regulated extracellular calcium-sensing via the CaR as a novel mediator of OA progression. PMID- 15882564 TI - A new biotechnology for articular cartilage repair: subchondral implantation of a composite of interconnected porous hydroxyapatite, synthetic polymer (PLA-PEG), and bone morphogenetic protein-2 (rhBMP-2). AB - OBJECTIVE: Articular cartilage repair remains a major obstacle in tissue engineering. We recently developed a novel tool for articular cartilage repair, consisting of a triple composite of an interconnected porous hydroxyapatite (IP CHA), recombinant human bone morphogenetic protein-2 (rhBMP-2), and a synthetic biodegradable polymer [poly-d,l-lactic acid/polyethylene glycol (PLA-PEG)] as a carrier for rhBMP-2. In the present study, we evaluated the capacity of the triple composite to induce the regeneration of articular cartilage. METHODS: Full thickness cartilage defects were created in the trochlear groove of 52 New Zealand White rabbits. Sixteen defects were filled with the bone morphogenetic protein (BMP)/PLA-PEG/IP-CHA composite (group I), 12 with PLA-PEG/IP-CHA (group II), 12 with IP-CHA alone (group III), and 12 were left empty (group IV). The animals were killed 1, 3, and 6 weeks after surgery, and the gross appearance of the defect sites was assessed. The harvested tissues were examined radiographically and histologically. RESULTS: One week after implantation with the BMP/PLA-PEG/IP-CHA composite (group I), vigorous repair had occurred in the subchondral defect. It contained an agglomeration of mesenchymal cells which had migrated from the surrounding bone marrow either directly, or indirectly via the interconnecting pores of the IP-CHA scaffold. At 6 weeks, these defects were completely repaired. The regenerated cartilage manifested a hyaline-like appearance, with a mature matrix and a columnar organization of chondrocytes. CONCLUSIONS: The triple composite of rhBMP-2, PLA-PEG, and IP-CHA promotes the repair of full-thickness articular cartilage defects within as short a period as 3 weeks in the rabbit model. Hence, this novel cell-free implant biotechnology could mark a new development in the field of articular cartilage repair. PMID- 15882565 TI - Degenerative knee joint disease in mice lacking 3'-phosphoadenosine 5' phosphosulfate synthetase 2 (Papss2) activity: a putative model of human PAPSS2 deficiency-associated arthrosis. AB - OBJECTIVE: Murine brachymorphism (bm) results from an autosomal recessive mutation of the Papss2 gene that encodes 3'-phosphoadenosine 5'-phosphosulfate synthetase 2, one of the principal enzymes required for the sulfation of extracellular matrix molecules in cartilage and other tissues. A spondyloepimetaphyseal dysplasia has been identified in Pakistani kindred having a mutation of PAPSS2. In addition to skeletal malformations that include short stature evident at birth due to limb shortening, brachydactyly, and kyphoscoliosis, affected individuals demonstrate premature onset degenerative joint disease. We investigated whether loss of Papss2 activity would similarly lead to degenerative joint disease in mice. METHODS: Mice carrying the bm mutation on a C57BL/6 background were obtained from the Jackson Laboratory. Limbs were analyzed by micro-computed tomography (microCT) and histology. RESULTS: At 12 months of age both male and female bm mice exhibited severe degenerative knee joint disease, with cartilage damage being primarily evident in the patello femoral and medial compartments. Control 12-14-month-old C57BL/6 mice, in contrast, only occasionally demonstrated minimal cartilage damage. muCT imaging of bm limbs revealed shortened diaphyses associated with flared metaphyses in the proximal elements of both fore and hind limbs. Additionally, the bm hind limbs demonstrated extensive structural alterations, characterized by distortion of the patello-femoral groove, and prominent bowing of both tibia and fibula. CONCLUSIONS: The bm mutant, which develops severe articular cartilage lesions of the knee joint by approximately 12 months of age, represents a novel example of murine degenerative joint disease, possibly representing a model of human PAPSS2 deficiency-associated arthrosis. PMID- 15882566 TI - Elucidation of IL-1/TGF-beta interactions in mouse chondrocyte cell line by genome-wide gene expression. AB - OBJECTIVE: To elucidate the antagonism between interleukin-1 (IL-1) and transforming growth factor-beta (TGF-beta) at the gene expression level, as IL-1 and TGF-beta are postulated to be critical mediators of cartilage degeneration/protection in rheumatic diseases. METHODS: The H4 chondrocyte cell line was validated by comparing metalloproteinase expression profile with intact murine cartilage by reverse transcription polymerase chain reaction. Genome-wide gene expression in the H4 cells in response to IL-1 and TGF-beta, alone and in combination, was analyzed by using oligonucleotide arrays negotiating approximately 12,000 genes. RESULTS: The response of cartilage and the H4 cell line to IL-1 and TGF-beta was comparable. Oligonucleotide array analysis demonstrated a mutual but asymmetrical antagonism as the dominant mode of interaction between IL-1 and TGF-beta. Cluster analysis revealed a remarkable selectivity in the mode of action exerted by TGF-beta on IL-1 regulated genes: antagonistic on pro-inflammatory genes whereas additive on growth regulators such as vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF). While the former cluster underlined the protective effect of TGF-beta, the latter underscored the adverse effect of TGF-beta. We further identified potentially novel classes of target genes under control of TGF-beta such as ras family, histones, proteasome components, and ubiquitin family, highlighting the importance of such genes in TGF signaling besides the well-characterized SMAD pathway. CONCLUSIONS: We identified a cluster of genes as potential targets mediating the adverse effect of TGF-beta such as fibrosis. Transcriptional regulation of ras GTPase and ubiquitin/proteasome pathways is likely to be a novel mechanism mediating the effect of TGF-beta and its interaction with IL-1. These down-stream genes and pathways can be targets in future therapy. PMID- 15882567 TI - Enhanced expression of insulin-like growth factor-binding proteins in human osteoarthritic cartilage detected by immunohistochemistry and in situ hybridization. AB - OBJECTIVE: To determine the roles of insulin-like growth factor (IGF) and IGF binding protein (IGFBP) in the pathogenesis of osteoarthritis (OA). DESIGN: Cartilage tissues were obtained from the femoral heads of patients with OA, and those from patients with femoral neck fractures were used as a control. The expression of IGFBP-3, -4, and -5 was examined using immunohistochemistry and in situ hybridization, and IGF-I and IGF-I receptors were also immunohistochemically detected. The percentages of positive chondrocytes were determined by counting the total number of chondrocytes over the area of the surface, middle, and deep zones of the cartilage. RESULTS: There was a marked increase in the percentage of positive chondrocytes in all IGFBPs on protein and messenger RNA levels for OA compared to that of the control cartilage. Furthermore, enhanced expression of IGFBPs and the IGF-I/IGF-I receptor was positively correlated with the histologic score for cartilage lesions. CONCLUSION: Up-regulation of IGFBPs as well as IGF-I and its receptor was observed for OA cartilage tissue, suggesting the involvement of IGFBPs in the pathogenesis of OA. PMID- 15882568 TI - Comparing different therapeutic classes for the treatment of osteoarthritis of the knee: data from the OMERACT-OARSI responder criteria analysis. PMID- 15882571 TI - Evolution of the brain developmental plan: Insights from agnathans. AB - In vertebrate evolution, the brain exhibits both conserved and unique morphological features in each animal group. Thus, the molecular program of nervous system development is expected to have experienced various changes through evolution. In this review, we discuss recent data from the agnathan lamprey (jawless vertebrate) together with available information from amphioxus and speculate the sequence of changes during chordate evolution that have been brought into the brain developmental plan to yield the current variety of the gnathostome (jawed vertebrate) brains. PMID- 15882573 TI - Cadherin 23 is a component of the transient lateral links in the developing hair bundles of cochlear sensory cells. AB - Cadherin 23 is required for normal development of the sensory hair bundle, and recent evidence suggests it is a component of the tip links, filamentous structures thought to gate the hair cells' mechano-electrical transducer channels. Antibodies against unique peptide epitopes were used to study the properties of cadherin 23 and its spatio-temporal expression patterns in developing cochlear hair cells. In the rat, intra- and extracellular domain epitopes are readily detected in the developing hair bundle between E18 and P5, and become progressively restricted to the distal tip of the hair bundle. From P13 onwards, these epitopes are no longer detected in hair bundles, but immunoreactivity is observed in the apical, vesicle-rich, pericuticular region of the hair cell. In the P2-P3 mouse cochlea, immunogold labeling reveals cadherin 23 is associated with kinocilial links and transient lateral links located between and within stereociliary rows. At this stage, the cadherin 23 ectodomain epitope remains on the hair bundle following BAPTA or La(3+) treatment, but is lost following exposure to the protease subtilisin. In contrast, mechano electrical transduction is abolished by BAPTA but unaffected by subtilisin. These results suggest cadherin 23 is associated with transient lateral links that have properties distinct from those of the tip-link. PMID- 15882572 TI - Blastocysts don't go it alone. Extrinsic signals fine-tune the intrinsic developmental program of trophoblast cells. AB - The preimplantation embryo floats freely within the oviduct and is capable of developing into a blastocyst independently of the maternal reproductive tract. While establishment of the trophoblast lineage is dependent on expression of developmental regulatory genes, further differentiation leading to blastocyst implantation in the uterus requires external cues emanating from the microenvironment. Recent studies suggest that trophoblast differentiation requires intracellular signaling initiated by uterine-derived growth factors and integrin-binding components of the extracellular matrix. The progression of trophoblast development from the early blastocyst stage through the onset of implantation appears to be largely independent of new gene expression. Instead, extrinsic signals direct the sequential trafficking of cell surface receptors to orchestrate the developmental program that initiates blastocyst implantation. The dependence on external cues could coordinate embryonic activities with the developing uterine endometrium. Biochemical events that regulate trophoblast adhesion to fibronectin are presented to illustrate a developmental strategy employed by the peri-implantation blastocyst. PMID- 15882574 TI - Spatiotemporal pattern and isoforms of cadherin 23 in wild type and waltzer mice during inner ear hair cell development. AB - Mutant alleles of the gene encoding cadherin 23 are associated with Usher syndrome type 1 (USH1D), isolated deafness (DFNB12) in humans, and deafness and circling behavior in waltzer (v) mice. Stereocilia of waltzer mice are disorganized and the kinocilia misplaced, indicating the importance of cadherin 23 for hair bundle development. Cadherin 23 was localized to developing stereocilia and proposed as a component of the tip link. We show that, during development of the inner ear, cadherin 23 is initially detected in centrosomes at E14.5, then along the length of emerging stereocilia, and later becomes concentrated at and subsequently disappears from the tops of stereocilia. In mature vestibular hair bundles, cadherin 23 is present along the kinocilium and in the region of stereocilia-kinocilium bonds, a pattern conserved in mammals, chicks, and frogs. Cadherin 23 is also present in Reissner's membrane (RM) throughout development. In homozygous v(6J) mice, a reported null allele, cadherin 23 was absent from stereocilia, but present in kinocilia, RM, and centrosomes. We reconciled these results by identifying two novel isoforms of Cdh23 unaffected in sequence and expression by the v(6J) allele. Our results suggest that Cdh23 participation in stereocilia links may be restricted to developing hair bundles. PMID- 15882575 TI - The homeodomain protein Meis1 is essential for definitive hematopoiesis and vascular patterning in the mouse embryo. AB - Homeodomain proteins of the Meis subfamily are expressed dynamically in several organs during embryogenesis and exert potent regulatory activity through their interaction with Hox proteins and other transcription factors. Here we show that Meis1 is expressed in the hematopoietic stem cell (HSC) compartment in the fetal liver, and in the primary sites of definitive hematopoiesis, including the aorta gonad-mesonephros (AGM) mesenchyme, the hemogenic embryonic arterial endothelium, and hematopoietic clusters within the aorta, vitelline, and umbilical arteries. We inactivated the Meis1 gene in mice and found that Meis1 mutant mice die between embryonic days 11.5 and 14.5, showing internal hemorrhage, liver hypoplasia, and anemia. In Meis1 mutant mouse fetal liver and AGM, HSC compartments are severely underdeveloped and colony-forming potential is profoundly impaired. AGM mesenchymal cells expressing Runx1, an essential factor for definitive HSC specification, are almost absent in mutant mice. In addition, hematopoietic clusters in the dorsal aorta, vitelline, and umbilical arteries are reduced in size and number. These results show a requirement for Meis1 in the establishment of definitive hematopoiesis in the mouse embryo. Meis1 mutant mice also displayed complete agenesis of the megakaryocyte lineage and localized defects in vascular patterning, which may cause the hemorrhagic phenotype. PMID- 15882576 TI - TGF-beta3 regulates anchoring junction dynamics in the seminiferous epithelium of the rat testis via the Ras/ERK signaling pathway: An in vivo study. AB - Recent studies have shown that transforming growth factor (TGF)-beta3 regulates blood-testis barrier (BTB) dynamics in vivo, plausibly by determining the steady state levels of occludin and zonula occludens-1 (ZO-1) at the BTB site via the p38 MAP kinase signaling pathway. Since BTB is composed of coexisting TJs and basal ectoplasmic specializations [ES, a testis-specific adherens junction (AJ) type] in the seminiferous epithelium of the rat testis, we sought to examine if TGF-beta3 would also regulate anchoring junction dynamics. Using an in vivo model in which rats were treated with AF-2364 [1-(2,4-dichlorobenzyl)-indazole-3 carbohydrazide] to perturb Sertoli-germ cell AJs without affecting the integrity of TJs at the BTB, it was noted that the event of germ cell loss from the epithelium was associated with a transient surge in TGF-beta3. Furthermore, it was also associated with a surge in the protein levels of Ras, p-ERK, and the intrinsic activity of ERK, illustrating TGF-beta3 apparently regulates Sertoli germ cell ES function via the Ras/MEK/ERK signaling pathway. Indeed, pretreatment of rats with TbetaRII/Fc chimera, a TGF-beta antagonist, or U0126, a specific MEK inhibitor, could significantly delay and partially block the disruptive effects of AF-2364 in depleting germ cells from the epithelium. While the protein levels of the cadherin/catenin complex were significantly induced during AF-2364 mediated germ cell loss, perhaps being used to retain germ cells in the epithelium, this increase failed to reverse the loss of adhesion function between Sertoli and germ cells because of a loss of protein-protein interactions between cadherins and catenins. Collectively, these results illustrate that the testis has a novel mechanism in place in which an agent that primarily disrupts TJs can induce secondary loss of AJ function, leading to germ cell loss from the seminiferous epithelium. Yet an agent that selectively disrupts AJs (e.g., AF 2364) can limit its effects exclusively at the Sertoli-germ cell adhesive site without perturbing the Sertoli-Sertoli TJs. PMID- 15882577 TI - Sox9 neural crest determinant gene controls patterning and closure of the posterior frontal cranial suture. AB - Cranial suture development involves a complex interaction of genes and tissues derived from neural crest cells (NCC) and paraxial mesoderm. In mice, the posterior frontal (PF) suture closes during the first month of life while other sutures remain patent throughout the life of the animal. Given the unique NCC origin of PF suture complex (analogous to metopic suture in humans), we performed quantitative real-time PCR and immunohistochemistry to study the expression pattern of the NCC determinant gene Sox9 and select markers of extracellular matrix. Our results indicated a unique up-regulated expression of Sox9, a regulator of chondrogenesis, during initiation of PF suture closure, along with the expression of specific cartilage markers (Type II Collagen and Type X Collagen), as well as cartilage tissue formation in the PF suture. This process was followed by expression of bone markers (Type I Collagen and Osteocalcin), suggesting endochondral ossification. Moreover, we studied the effect of haploinsufficiency of the NCC determinant gene Sox9 in the NCC derived PF suture complex. A decrease in dosage of Sox9 by haploinsufficiency in NCC-derived tissues resulted in delayed PF suture closure. These results demonstrate a unique development of the PF suture complex and the role of Sox9 as an important contributor to timely and proper closure of the PF suture through endochondral ossification. PMID- 15882578 TI - molting defective is required for ecdysone biosynthesis. AB - 20-hydroxyecdysone was discovered as the major biologically active insect steroid hormone half a century ago, yet much remains to be learned about its biosynthesis and its activities. 20-hydroxyecdysone controls many biological processes, including progression between larval stages, entry to pupariation and metamorphosis. A number of genes required for 20-hydroxyecdysone production have been identified, including those encoding enzymes that mediate four of the late steps of biosynthesis. A second smaller group of low ecdysone mutants do not encode enzymes. Here, we report identification of one such gene, which we call molting defective, on the basis of its lethal phenotype. molting defective encodes a nuclear zinc finger protein required for ecdysone biosynthesis. PMID- 15882579 TI - Semaphorin signaling guides cranial neural crest cell migration in zebrafish. AB - Cranial neural crest cells (NCCs) migrate into the pharyngeal arches in three primary streams separated by two cranial neural crest (NC)-free zones. Multiple tissues have been implicated in the guidance of cranial NCC migration; however, the signals provided by these tissues have remained elusive. We investigate the function of semaphorins (semas) and their receptors, neuropilins (nrps), in cranial NCC migration in zebrafish. We find that genes of the sema3F and sema3G class are expressed in the cranial NC-free zones, while nrp2a and nrp2b are expressed in the migrating NCCs. sema3F/3G expression is expanded homogeneously in the head periphery through which the cranial NCCs migrate in lzr/pbx4 mutants, in which the cranial NC streams are fused. Antisense morpholino knockdown of Sema3F/3G or Nrp2 suppresses the abnormal cranial NC phenotype of lzr/pbx4 mutants, demonstrating that aberrant Sema3F/3G-Nrp2 signaling is responsible for this phenotype and suggesting that repulsive Sema3F/3G-Npn2 signaling normally contributes to the guidance of migrating cranial NCCs. Furthermore, global over expression of sema3Gb phenocopies the aberrant cranial NC phenotype of lzr/pbx4 mutants when endogenous Sema3 ligands are knocked down, consistent with a model in which the patterned expression of Sema3 ligands in the head periphery coordinates the migration of Nrp-expressing cranial NCCs. PMID- 15882580 TI - Id4 is required for the correct timing of neural differentiation. AB - Complex intrinsic and extrinsic mechanisms determine neural cell fate during development of the nervous system. Using Id4 deficient mice, we show that Id4 is required for normal development of the central nervous system (CNS), timing neural differentiation in the developing forebrain. In the absence of Id4, the ventricular zone of the neocortex, future hippocampus as well as lateral and medial ganglionic eminences exhibited a 20-30% reduction in mitotic neural precursor cells (NPCs). Although the number of apoptotic cells was significantly increased, the neocortex of Id4(-/-) embryos was consistently thicker due to premature neuronal differentiation, which resulted in an increase in early-born neurons in the adult Id4(-/-) cortex. Late-born cortical neurons and astrocytes in the cortex, septum, hippocampus and caudate putamen of Id4(-/-) adult brains were decreased, however, likely due to the depletion of the NPC pool. Consequently, adult Id4(-/-) brains were smaller and exhibited enlarged ventricles. In vitro analysis of neurosphere cultures revealed that proliferation of Id4-deficient NPCs was impaired and that BMP2-mediated astrocyte differentiation was accelerated in the absence of Id4. Together, these in vivo and in vitro data suggest a crucial role for Id4 in regulating NPC proliferation and differentiation. PMID- 15882581 TI - Insertion of Cre into the Pax3 locus creates a new allele of Splotch and identifies unexpected Pax3 derivatives. AB - Pax3 is a transcription factor expressed in the dorsal neural tube and somite of the developing embryo. It plays critical roles in pre-migratory neural crest cells and in myogenic precursors of skeletal muscle. Pax3-deficient Splotch embryos display neural tube and neural crest defects and lack hypaxial muscles. We have created a new allele of Splotch by replacing the first coding exon with a gene encoding Cre recombinase. This functions as a null allele and no Pax3 protein is detected in homozygous embryos. Heterozygous Pax3(Cre/+) mice display a white belly spot, as do Splotch heterozygotes. Homozygous Pax3(Cre/Cre) embryos are embryonic lethal. We have used Pax3(Cre/+) mice to fate-map Pax3 derivatives in the developing mouse. As expected, neural crest and some somitic derivatives are identified. However, we also detect previously unappreciated derivatives of Pax3-expressing precursors in the colonic epithelium of the hindgut and within the urogenital system. PMID- 15882582 TI - The lesswright mutation activates Rel-related proteins, leading to overproduction of larval hemocytes in Drosophila melanogaster. AB - The lesswright (lwr) gene encodes an enzyme that conjugates a small ubiquitin related modifier (SUMO). Since the conjugation of SUMO occurs in many different proteins, a variety of cellular processes probably require lwr function. Here, we demonstrate that lwr function regulates the production of blood cells (hemocytes) in Drosophila larvae. lwr mutant larvae develop many melanotic tumors in the hemolymph at the third instar stage. The formation of melanotic tumors is due to a large number of circulating hemocytes, which is approximately 10 times higher than those of wild type. This overproduction of hemocytes is attributed to the loss of lwr function primarily in hemocytes and the lymph glands, a hematopoietic organ in Drosophila larvae. High incidences of Dorsal (Dl) protein in the nucleus were observed in lwr mutant hemocytes, and the dl and Dorsal-related immunity factor (Dif) mutations were found to be suppressors of the lwr mutation. Therefore, the lwr mutation leads to the activation of these Rel-related proteins, key transcription factors in hematopoiesis. We also demonstrate that dl and Dif play different roles in hematopoiesis. dl primarily stimulates plasmatocyte production, but Dif controls both plasmatocyte and lamellocyte production. PMID- 15882583 TI - Retinoic acid influences the development of the inferior olivary nucleus in the rodent. AB - All-trans retinoic acid (atRA) is an endogenous morphogen that regulates gene transcription. Maternal exposure to atRA results in severe developmental abnormalities by disrupting normal patterns of atRA distribution. Previously, we have shown that the pontine nucleus, which originates from the rhombic lip, is severely atrophied in the mouse on exposure to atRA at gestational days 9 and 10. In this study, we show that this same period of atRA exposure has the contrary effect on the inferior olive and this rhombic lip derivative is expanded in volume and probably contains an increased number of cells. The posterior region of the inferior olive maintains a relatively normal shape but is significantly expanded in size. In contrast, the organization of the anterior inferior olive is severely disrupted. Because endogenous atRA levels are known to be higher in the region of the posterior inferior olive at the time of birth of inferior olivary neurons, these results suggest that endogenous atRA may promote the generation, or select the fate, of posterior neurons of the inferior olive. In support of this concept, a reduction in atRA resulting from vitamin A deficiency results in loss of cells of the posterior inferior olive. PMID- 15882584 TI - Strongylocentrotus purpuratus transcription factor GATA-E binds to and represses transcription at an Otx-Goosecoid cis-regulatory element within the aboral ectoderm-specific spec2a enhancer. AB - During Strongylocentrotus purpuratus embryogenesis, aboral ectoderm-specific expression of spec2a relies on an upstream enhancer that confers its spatial specificity largely through repression. The purpose of this study was to determine how spec2a expression is repressed in endoderm and oral ectoderm territories. A 78-base pair DNA sequence within the enhancer contains five tightly spaced cis-regulatory elements including proximal (TAATCT) and distal (TAATCC) elements that bind to both SpOtx, a broadly distributed transcriptional activator, and SpGoosecoid (SpGsc), an oral ectoderm-restricted transcriptional repressor. We show here that these two seemingly redundant Otx/Gsc elements have distinct functions. The proximal element bound to SpGATA-E, an endomesoderm specific transcription factor. Treatment with SpGATA-E and SpGsc morpholino antisense oligonucleotides (MASOs) resulted in enhanced transcriptional activity from the proximal element, suggesting that both factors functioned as repressors at this site. SpGATA-E MASO-treated embryos failed to express ectoderm markers, indicating a role for SpGATA-E in ectoderm differentiation. The spec2a proximal element was distinct from the corresponding element in the related spec1 enhancer, and swaps between spec1 and spec2a cis-regulatory elements indicated, that for optimal repression, the proximal element had to interact with a nearby CCAAT-binding factor element. Our results show that the recently evolved proximal element contributes to the repression of spec2a in endomesoderm and oral ectoderm territories. PMID- 15882585 TI - Subversion of T lineage commitment by PU.1 in a clonal cell line system. AB - Specification of mammalian T lymphocytes involves prolonged developmental plasticity even after lineage-specific gene expression begins. Expression of transcription factor PU.1 may maintain some myeloid-like developmental alternatives until commitment. Commitment could reflect PU.1 shutoff, resistance to PU.1 effects, and/or imposition of a suicide penalty for diversion. Here, we describe subclones from the SCID.adh murine thymic lymphoma, adh.2C2 and adh.6D4, that represent a new tool for probing these mechanisms. PU.1 can induce many adh.2C2 cells to undergo diversion to a myeloid-like phenotype, in an all-or-none fashion with multiple, coordinate gene expression changes; adh.6D4 cells resist diversion, and most die. Diversion depends on the PU.1 Ets domain but not on known interactions in the PEST or Q-rich domains, although the Q-rich domain enhances diversion frequency. Protein kinase C/MAP kinase stimulation can make adh.6D4 cells permissive for diversion without protecting from suicide. These results show distinct roles for regulated cell death and another stimulation sensitive function that establishes a threshold for diversion competence. PU.1 also diverts normal T-cell precursors from wild type or Bcl2-transgenic mice to a myeloid-like phenotype, upon transduction in short-term culture. The adh.2C2 and adh.6D4 clones thus provide an accessible system for defining mechanisms controlling developmental plasticity in early T-cell development. PMID- 15882586 TI - A gene network downstream of transcription factor Math5 regulates retinal progenitor cell competence and ganglion cell fate. AB - Math5, a mouse homolog of the Drosophila proneural bHLH transcription factor Atonal, is essential in the developing retina to establish retinal progenitor cell competence for a ganglion cell fate. Elucidating the mechanisms by which Math5 influences progenitor cell competence is crucial for understanding how specification of neuronal cell fate occurs in the retina and it requires knowledge of the downstream target genes that depend on Math5 for their expression. To date, only a handful of genes downstream of Math5 have been identified. To better define the gene network operating downstream of Math5, we used custom-designed microarrays to examine the changes in embryonic retinal gene expression caused by deletion of math5. We identified 270 Math5-dependent genes, including those that were expressed specifically either in progenitor cells or differentiated ganglion cells. The ganglion cell-specific genes included both Brn3b-dependent and Brn3b-independent genes, indicating that Math5 regulates distinct branches of the gene network responsible for retinal ganglion cell differentiation. In math5-null progenitor cells, there was an up-regulation of the proneural genes math3, neuroD, and ngn2, indicating that Math5 suppresses the production of other cell types in addition to promoting retinal ganglion cell formation. The promoter regions of many Math5-dependent genes contained binding sites for REST/NRSF, suggesting that release from general repression in retinal progenitor cells is required for ganglion cell-specific gene activation. The identification of multiple roles for Math5 provides new insights into the gene network that defines progenitor cell competence in the embryonic retina. PMID- 15882587 TI - Localized repressors delineate the neurogenic ectoderm in the early Drosophila embryo. AB - The Dorsal gradient produces sequential patterns of gene expression across the dorsoventral axis of early embryos, thereby establishing the presumptive mesoderm, neuroectoderm, and dorsal ectoderm. Spatially localized repressors such as Snail and Vnd exclude the expression of neurogenic genes in the mesoderm and ventral neuroectoderm, respectively. However, no repressors have been identified that establish the dorsal limits of neurogenic gene expression. To investigate this issue, we have conducted an analysis of the ind gene, which is selectively expressed in lateral regions of the presumptive nerve cord. A novel silencer element was identified within the ind enhancer that is essential for eliminating expression in the dorsal ectoderm. Evidence is presented that the associated repressor can function over long distances to silence neighboring enhancers. The ind enhancer also contains a variety of known activator and repressor elements. We propose a model whereby Dorsal and EGF signaling, together with the localized Schnurri repressor, define a broad domain of ind expression throughout the entire presumptive neuroectoderm. The ventral limits of gene expression are defined by the Snail and Vnd repressors, while the dorsal border is established by the newly defined silencer element. PMID- 15882588 TI - Convergent genetic programs regulate similarities and differences between related motor neuron classes in Caenorhabditis elegans. AB - How do genetic programs create features common to a specific cell or tissue type while generating modifications necessary for functional diversification? We have addressed this question using the nematode Caenorhabditis elegans. The dorsal D (DD) and ventral D (VD) motorneurons (mns), referred to collectively as the D mns, compose a cross-inhibitory network that contributes to the animal's sinuous locomotion. The D mns share a number of structural and functional features, but are distinguished from one another by their synaptic patterns and the expression of a neuropeptide gene. Our findings suggest that the similarities and differences are generated at the transcriptional level. UNC-30 contains a homeodomain and activates structural and functional genes expressed in both classes. UNC-55 is a nuclear receptor expressed in the VD mns that is necessary for generating features that distinguish the two classes of D mns from one another. In unc-55 mutants, the VD mns adopt the DD mn synaptic pattern and peptide expression profile. Conversely, ectopic expression of unc-55 in the DD mns causes them to adopt VD mn features. The promoter of the neuropeptide gene expressed in the DD mns contains putative binding sites for both UNC-30 and UNC 55; alteration of these sites suggests that UNC-55 represses the ability of UNC 30 to activate a subset of genes that are expressed in the DD mns but not in the VD mns. Thus UNC-55 acts as a switch for the features that distinguish these two functionally related classes of mns. PMID- 15882589 TI - The role of ARGONAUTE1 (AGO1) in meristem formation and identity. AB - The ARGONAUTE gene family is involved in the regulation of gene expression via the RNAi Silencing Complex (RISC). microRNA (miRNA) are 20-22bp RNAs that direct RISC to target genes. Several miRNA have been characterized in plants. Their roles include control of flowering time, floral organ identity, cell division patterns, and leaf polarity. ARGONAUTE1 (AGO1) is required for stem cell function and organ polarity, as is the closely related protein PINHEAD/ZWILLE (PNH/ZLL). Through phenotypic and double mutant analysis, we show that AGO1 regulates stem cell function via SHOOT MERISTEMLESS (STM). CUPSHAPED COTYLEDONS1 and 2 (CUC1 and CUC2) positively regulate STM and are targets of miRNA. The effect of AGO1 on leaf polarity is dependent, in part, on its role in meristem function revealed by interactions with ASYMMETRIC LEAVES1(AS1). AGO1 is required for full expression of LEAFY (LFY), APETALA1 (AP1) and AGAMOUS (AG). Flowering time is unaffected but floral meristem identity is partially restored in a curlyleaf (clf) background and this is not due to clf's affects on AG expression. CLF is over expressed in ago1, showing that the RNAi pathway regulates polycomb-type epigenetic modifiers. PMID- 15882591 TI - History of flexor tendon repair. AB - The first issue of Hand Clinics published 20 years ago was devoted to flexor tendon injuries. This was most appropriate, because no subject in hand surgery has sparked more interest or discussion. That inaugural issue included excellent presentations on the basic science of tendon injuries (anatomy, biomechanics, nutrition, healing, adhesions) and the clinical practice of tendon repair. Of interest, there was no presentation on the fascinating history of flexor tendon surgery. It is most appropriate, therefore, that this current update of the flexor tendon begins with a historical review of the evolution of flexor tendon repair. PMID- 15882592 TI - Biomechanics of the flexor tendons. AB - This article examines basic tendon biomechanics, the anatomy and mechanics of digital flexor tendons, and the digital flexor pulley system. It also explores the various models that have tried to simulate the motion of the flexor tendons and several testing modalities that have been used. Finally, clinical applications are considered, including the biomechanics of flexor tendon repairs and tendon transfers. As we reach limits in the care of flexor tendon injuries, research into molecular, biochemical, and micromechanical methods of tendon repair will become the forefront of future investigation. PMID- 15882593 TI - Flexor tendons: anatomy and surgical approaches. AB - The nature of the original injury is the chief determinant of outcome and is out of the control of the surgeon. Every step thereafter can be influenced by him or her, though, and a thorough knowledge of anatomy and surgical approaches allows for the best possible repair under any set of circumstances. PMID- 15882594 TI - Flexor tendon biology. AB - Significant advances in the understanding of intrasynovial flexor tendon repair and rehabilitation have been made since the early 1970s. The concept of adhesion free, or primary tendon healing--that tendons could heal intrinsically without the ingrowth of fibrous adhesions from the surrounding sheath has been validated both experimentally and clinically in studies over the past 25 years. Recent attempts to understand and improve the results of intrasynovial flexor tendon repair have focused upon restoration of the gliding surface, augmentation of early post-operative repair site biomechanical strength and on the elucidation of the molecular biology of early post-operative tendon healing. The goals of the surgical treatment of patients with intrasynovial flexor tendon lacerations remain unchanged: to achieve a primary tendon repair of sufficient tensile strength to allow application of a post-operative mobilization rehabilitation protocol. This program should inhibit the formation of intrasynovial adhesions and restore the gliding surface, while facilitating the healing of the repair site. PMID- 15882595 TI - Zone I flexor tendon injuries. AB - Zone I flexor tendon injuries entail injuries to the flexor digitorum profundus (FDP) tendon. These injuries occur distal to the superficialis insertion over the middle phalanx or proximal distal phalanx, and as such are isolated injuries to the FDP. The mechanism most commonly is closed avulsion from the distal phalanx or a laceration, but other mechanisms such as open avulsion or crush injury can occur. On physical examination, the cascade of the fingers will be disrupted, and distal interphalangeal (DIP) joint flexion must be isolated to determine if the FDP tendon is continuous, as other mechanisms are present to enable finger flexion at the other joints. PMID- 15882596 TI - Acute flexor tendon repairs in zone II. AB - Flexor tendon repair in zone II is still a technically demanding procedure, but the outcomes have become more predictable and satisfying. Of keystone importance for obtaining the goals of normal strength and gliding of repaired flexor tendons are an atraumatic surgical technique, an appropriate suture material, a competent pulley system, and the use of early motion rehabilitation protocols. The overall goal of hand and finger function also implies timely addressing of neurovascular injuries. New devices such as the TenoFix (Ortheon Medical; Winter Park, Florida) have shown adequate strength in the laboratory but are bulky and untested for work of flexion. Insufficient clinical data and high cost may prevent widespread use. PMID- 15882597 TI - Treatment of acute flexor tendon injury: zones III-V. AB - Many of the principles of flexor tendon repair and rehabilitation can be applied to zones III-V. Injuries in zones III-V are rarely isolated and neurovascular involvement is common. Because of the often extensive and unknown degree of injury, there should be a low threshold for surgical wound exploration. Primary repair of injured tendons and neurovascular structures is recommended by way of a systematic approach. Good to excellent outcomes in range of motion and tendon function can be expected; however, functional outcomes of associated nerve injuries are varied, with younger patients generally demonstrating the best results (Fig. 2E). PMID- 15882598 TI - Complex injuries including flexor tendon disruption. AB - The treatment of tendon injury in combined complex injuries to the hand is dictated by the presence of concomitant injuries. Early range of motion is desirable. To achieve this, fractures must be stabilized and the soft tissue envelope and vascular integrity maintained or reconstituted. In those instances in which these conditions cannot be met, the surgeon and patient should be prepared for secondary surgeries, including reconstruction or tenolysis. Although nerve integrity is not necessary for early functional success following tenorrhaphy, nerve injuries should be repaired or grafted primarily as the injury permits. In cases in which vascular compromise is encountered, the options of revascularization versus primary amputation should be discussed with the patient. With an understanding of the treatment principles, the complications associated with complex tendon injuries can be minimized. It is important to stress that optimal functional outcome is multifactorial and includes a physician-therapist team-oriented approach. PMID- 15882599 TI - Clinical outcomes associated with flexor tendon repair. AB - Review of the outcomes of clinical flexor tendon repairs reported over the past 15 years showed advances in the outcomes with excellent or good functional return in more than three fourths of primary tendon repairs following a variety of postoperative passive/active mobilization treatments. Strickland and Glogovac criteria are the most commonly adopted methods to assess function. Repair ruptures (4%-10% for zone II finger flexors and 3%-17% for the FPL tendon), adhesion formations, and stiffness of finger joints remain frustrating problems in flexor tendon repairs and rehabilitation. Four approaches are suggested to improve outcomes of the repairs and to solve these difficult problems,which include stronger surgical repairs, appropriate pulleys or sheath management, optimization of rehabilitation regimens, and modern biologic approaches. PMID- 15882600 TI - Flexor tenolysis. AB - Flexor tenolysis is a challenging procedure with valuable clinical usefulness in the restitution and enhancement of digital function in the appropriate patient. In the absence of complications, improvement in digital flexion can be expected. The requisites for success are a skilled surgeon, a motivated and well informed patient, and a closely monitored hand therapy program. PMID- 15882601 TI - Delayed treatment of flexor tendon injuries including grafting. AB - This article synthesizes an enormous amount of peer reviewed articles, book chapters, and anecdotal clinical information regarding the late management of flexor tendon injuries by free-tendon grafting, tenolysis, and staged reconstruction. Some of the most pertinent historical contributions to these subjects have been reviewed in concert with an update regarding the most widely used current clinical methods for performing these procedures. This article points out areas of controversy and references the dissenting opinions from those presented here. The delayed treatment of flexor tendon injuries has advanced considerably in the last half century. Although much of the sage advice of the historic masters of flexor tendon surgery remains clinically applicable today, newer techniques and much improved therapy protocols have improved appreciably the results of the procedures described in this treatise. It is realistic and exciting to anticipate that the future will continue to improve the results of these methods through biologic manipulation of tendon healing and adhesion formation. PMID- 15882602 TI - Flexor tendon pulley reconstruction. AB - Pulley reconstruction remains a challenging intellectual and technical exercise. When performed correctly, however, it can be a gratifying procedure that provides much improved function of the digit. As described in this article, there are many different techniques by which the pulley can be reconstructed. Each of these techniques has distinct advantages and disadvantages. The hand surgeon should be familiar with each of these techniques and the general principles of pulley reconstruction as laid out in this article. With this knowledge base, the treating surgeon is able to tailor the procedure performed to the exact anatomy and clinical situation of each patient. Although great strides have been made over the past half century, more clinical research is needed to determine the best technique, not just in the cadaver model, but also in the complex model of the living human hand. PMID- 15882603 TI - Pediatric flexor tendon injuries. AB - Flexor tendon injuries in children differ from adults in their diagnosis and postoperative rehabilitation principles. The child may be uncooperative, so indirect methods of tendon integrity must be used for diagnosis. Radiographs may be useful for associated fracture or retained foreign bodies. A high index of suspicion necessitates surgical exploration. Although surgical approach and repair techniques are identical to those in adults, postoperative immobilization for 3-4 weeks is used instead of an early motion protocol. Delayed diagnosis is more common in the pediatric population, and recognition and management of postoperative complications can be difficult,because the child may be unable to cooperate or comply with the treatment. PMID- 15882604 TI - Rehabilitation after flexor tendon repair, reconstruction, and tenolysis. AB - Flexor tendon rehabilitation after injury and surgical intervention has progressed over the last several decades. This evolution has left a vast amount of information for the hand therapy clinician. The hand therapist treating a primary flexor tendon repair can easily feel daunted, confused, and apprehensive because of the sheer amount of information before him or her, which may lead to patient treatment with a textbook or cookbook approach. This article outlines the history of flexor tendon programs and their evidenced-based development so that the clinician can approach each patient individually and progress them with a personalized, tailored approach in close communication with the surgeon. Successful flexor tendon rehabilitation's end-result is functional hand motion and strength. As experimental studies on improved surgical techniques continue to develop, more clinical research to support rehabilitation techniques that lead to good hand function results are necessary. PMID- 15882605 TI - The future of flexor tendon surgery. AB - Clinical outcomes following flexor tendon repair have made significant improvements in the last 50 years. In that time standard treatment has evolved from secondary grafting to primary repair with postoperative rehabilitation protocols. Unfortunately, excellent results are not yet attained universally following treatment. Improving understanding of tendon healing at the cellular, molecular, and genetic levels will likely enable surgeons to modulate the normal repair process. We now look toward biologic augmentation of flexor tendon repairs to address the problems of increasing tensile strength while reducing adhesion formation following injury and operative repair. PMID- 15882606 TI - Interference with HH-GLI signaling inhibits prostate cancer. AB - The Hedgehog-Gli (Hh-Gli) signaling pathway controls many aspects of tissue patterning, cell proliferation, differentiation and regeneration and regulates cell number in various organs. In adults, the Hh-Gli pathway remains active in a number of stem cells and regenerating tissues. Inappropriate and uncontrolled HH GLI pathway activation has been demonstrated in a variety of human cancers. Three recent papers show that components of the pathway are expressed in human prostate tumors and, more importantly, that prostate cancers depend on sustained HH-GLI signaling. These data raise the possibility of a new therapeutic approach to treat this often lethal disease. PMID- 15882607 TI - CCL3L1 dose and HIV-1 susceptibility. AB - Several genetic factors influence HIV-1 susceptibility or AIDS disease progression. A recent study reported on what could be a particularly important genetic determinant for HIV-1 susceptibility and disease progression: copy number of a chemokine gene termed CCL3L1. Individuals with low copy numbers of the gene, relative to their ethnic background, were associated with markedly enhanced HIV 1/AIDS susceptibility. These findings define an important new genetic determinant of HIV-1 susceptibility and further emphasize the importance of the chemokine system, either as elements that inhibit HIV-1 infection or that modulate antiviral immune responses. PMID- 15882608 TI - Changing the logic of therapeutic angiogenesis for ischemic disease. AB - The worldwide epidemic of ischemic disease urgently requires innovative treatments. Recently, therapeutic angiogenesis has emerged as a noninvasive supply-side approach, aimed at promoting neovascularization in underperfused tissues through the local delivery of angiogenic growth factors. Successful preclinical studies paved the way for the first clinical trials, with single growth factors given as recombinant proteins or genes. However, clinical results have not matched the initial promises. Our opinion is that the logic of therapeutic angiogenesis needs profound revision. Here, we introduce the concept that pleiotropic agents can stimulate the healing of all the components of ischemic tissue. We also propose prophylactic interventions to delay vascular senescence. The optimization of therapeutic angiogenesis will open unprecedented opportunities for the care of life-threatening ischemic disease. PMID- 15882609 TI - Genes contributing to risk for common forms of stroke. AB - The quest for disease genes that confer risk for stroke is now being undertaken using three complementary approaches. Positional cloning using rare Mendelian phenocopies of stroke has found genes that contribute to rare forms of stroke but, so far, not to the common forms of stroke. Candidate-gene case-control association studies using the common forms of stroke have found suggestive associations of modest effect. However, positional cloning using hundreds of Icelandic families affected by the common forms of stroke has recently found two genes conferring substantial risk for ischemic stroke that have apparently been confirmed in the USA and other European populations. Both genes encode enzymes, phosphodiesterase 4D (PDE4D) and arachidonate 5-lipoxygenase-activating protein (FLAP), which suggest novel treatment strategies for stroke prevention. PMID- 15882610 TI - Immunotherapy: target the stroma to hit the tumor. AB - For decades it has been assumed that T cells reject tumors essentially by direct killing. However, solid tumors are composed of malignant cells and a variety of different nonmalignant cells, referred to as tumor stroma. Stromal cells, such as endothelial cells, fibroblasts and inflammatory cells, often support tumor growth. Here, we discuss new findings showing that the tumor stroma is an important target during T-cell-mediated tumor rejection. Cytotoxic molecules and cytokines produced by T cells inhibit or destroy the stromal 'infrastructure', thereby withdrawing essential resources and leading to tumor infarction and subsequent T-cell-mediated elimination of residual tumor cells. These findings are important for the development of more effective and specific immunotherapies for cancer. PMID- 15882611 TI - Therapeutic promise of JNK ATP-noncompetitive inhibitors. AB - Protein kinases are being increasingly targeted in the quest for new therapeutics, and the c-Jun N-terminal kinases (JNKs) are no exception. Protein kinase inhibitors are generally small molecules that show competitive inhibition with respect to ATP. However, a peptide has been developed that is an ATP noncompetitive inhibitor of JNK. This article describes the use of this peptide in an increasing number of animal models of disease, including diabetes, stroke, neurotrauma, hearing loss and Alzheimer's disease. The efficacy of this peptide shows that JNK inhibition is an effective strategy for the treatment of these diseases and opens the possibility for testing whether JNK inhibition will be beneficial in other diseases, such as atherosclerosis, arthritis and a range of neurodegenerative diseases. PMID- 15882612 TI - Genetic therapies for cardiovascular diseases. AB - Recent advances in understanding the molecular and cellular basis of cardiovascular diseases, together with the availability of tools for genetic manipulation of the cardiovascular system, offer possibilities for new treatments. Gene therapies have demonstrated potential usefulness for treating complex cardiovascular diseases, such as hypertension, atherosclerosis and myocardial ischemia, in various animal models. Some of these experimental therapies are now undergoing clinical evaluation in patients with cardiovascular disease. However, the successful transition of these therapies into mainstream clinical practice awaits further improvements to vector platforms and delivery tools and the documentation of clinical feasibility, safety and efficacy through multi-center randomized trials. PMID- 15882613 TI - Artificial and engineered chromosomes: non-integrating vectors for gene therapy. AB - Non-integrating gene-delivery platforms demonstrate promise as potentially ideal gene-therapy vector systems. Although several approaches are under development, there is little consensus as to what constitutes a true 'artificial' versus an 'engineered' human chromosome. Recent progress must be evaluated in light of significant technical challenges that remain before such vectors achieve clinical utility. Here, we examine the principal classes of non-integrating vectors, ranging from episomes to engineered mini-chromosomes to true human artificial chromosomes. We compare their potential as practical gene-transfer platforms and summarize recent advances towards eventual applications in gene therapy. Although chromosome-engineering technology has advanced considerably within recent years, difficulties in establishing composition of matter and effective vector delivery currently prevent artificial or engineered chromosomes being accepted as viable gene-delivery platforms. PMID- 15882614 TI - Translational repression by Bicoid: competition for the cap. AB - Eggs and embryos have proven to be fertile grounds for discovery and characterization of a variety of mechanisms used in posttranscriptional control of gene expression, mechanisms that are central to embryonic patterning and development. In this issue of Cell, solve a puzzle surrounding the action of the Drosophila Bicoid morphogen in formation of the Caudal protein gradient and in doing so describe a novel mechanism of translational repression. PMID- 15882615 TI - Cre-ating somatic cell genetic mosaics in the mouse. AB - Generation of somatic mosaics in which mutant cell clones are uniquely and completely labeled has yielded considerable insight into many biological processes in Drosophila. In this issue of Cell, describe a novel method called MADM that allows the generation of such mosaics in mice. PMID- 15882616 TI - Mitotic-exit control as an evolved complex system. AB - The exit from mitosis is the last critical decision during a cell-division cycle. A complex regulatory system has evolved to evaluate the success of mitotic events and control this decision. Whereas outstanding genetic work in yeast has led to rapid discovery of a large number of interacting genes involved in the control of mitotic exit, it has also become increasingly difficult to comprehend the logic and mechanistic features embedded in the complex molecular network. Our view is that this difficulty stems in part from the attempt to explain mitotic-exit control using concepts from traditional top-down engineering design, and that exciting new results from evolutionary engineering design applied to networks and electronic circuits may lend better insights. We focus on four particularly intriguing features of the mitotic-exit control system and attempt to examine these features from the perspective of evolutionary design and complex system engineering. PMID- 15882617 TI - Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. AB - Fibroblasts often constitute the majority of the stromal cells within a breast carcinoma, yet the functional contributions of these cells to tumorigenesis are poorly understood. Using a coimplantation tumor xenograft model, we demonstrate that carcinoma-associated fibroblasts (CAFs) extracted from human breast carcinomas promote the growth of admixed breast carcinoma cells significantly more than do normal mammary fibroblasts derived from the same patients. The CAFs, which exhibit the traits of myofibroblasts, play a central role in promoting the growth of tumor cells through their ability to secrete stromal cell-derived factor 1 (SDF-1); CAFs promote angiogenesis by recruiting endothelial progenitor cells (EPCs) into carcinomas, an effect mediated in part by SDF-1. CAF-secreted SDF-1 also stimulates tumor growth directly, acting through the cognate receptor, CXCR4, which is expressed by carcinoma cells. Our findings indicate that fibroblasts within invasive breast carcinomas contribute to tumor promotion in large part through the secretion of SDF-1. PMID- 15882618 TI - Transition from nonspecific to specific DNA interactions along the substrate recognition pathway of dam methyltransferase. AB - DNA methyltransferases methylate target bases within specific nucleotide sequences. Three structures are described for bacteriophage T4 DNA-adenine methyltransferase (T4Dam) in ternary complexes with partially and fully specific DNA and a methyl-donor analog. We also report the effects of substitutions in the related Escherichia coli DNA methyltransferase (EcoDam), altering residues corresponding to those involved in specific interaction with the canonical GATC target sequence in T4Dam. We have identified two types of protein-DNA interactions: discriminatory contacts, which stabilize the transition state and accelerate methylation of the cognate site, and antidiscriminatory contacts, which do not significantly affect methylation of the cognate site but disfavor activity at noncognate sites. These structures illustrate the transition in enzyme-DNA interaction from nonspecific to specific interaction, suggesting that there is a temporal order for formation of specific contacts. PMID- 15882619 TI - X-ray structures of the Sulfolobus solfataricus SWI2/SNF2 ATPase core and its complex with DNA. AB - SWI2/SNF2 ATPases remodel chromatin or other DNA:protein complexes by a poorly understood mechanism that involves ATP-dependent DNA translocation and generation of superhelical torsion. Crystal structures of a dsDNA-translocating SWI2/SNF2 ATPase core from Sulfolobus solfataricus reveal two helical SWI2/SNF2 specific subdomains, fused to a DExx box helicase-related ATPase core. Fully base paired duplex DNA binds along a central cleft via both minor groove strands, indicating that SWI2/SNF2 ATPases travel along the dsDNA minor groove without strand separation. A structural switch, linking DNA binding and the active site DExx motif, may account for the stimulation of ATPase activity by dsDNA. Our results suggest that torque in remodeling processes is generated by an ATP-driven screw motion of DNA along the active site cleft. The structures also redefine SWI2/SNF2 functional motifs and uncover unexpected structural correlation of mutations in Cockayne and X-linked mental retardation syndromes. PMID- 15882620 TI - Acetylation in histone H3 globular domain regulates gene expression in yeast. AB - In Saccharomyces cerevisiae, known histone acetylation sites regulating gene activity are located in the N-terminal tails protruding from the nucleosome core. We report lysine 56 in histone H3 as a novel acetylation site that is located in the globular domain, where it extends toward the DNA major groove at the entry exit points of the DNA superhelix as it wraps around the nucleosome. We show that K56 acetylation is enriched preferentially at certain active genes, such as those coding for histones. SPT10, a putative acetyltransferase, is required for cell cycle-specific K56 acetylation at histone genes. This allows recruitment of the nucleosome remodeling factor Snf5 and subsequent transcription. These findings indicate that histone H3 K56 acetylation at the entry-exit gate enables recruitment of the SWI/SNF nucleosome remodeling complex and so regulates gene activity. PMID- 15882621 TI - UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex. AB - The xeroderma pigmentosum group C (XPC) protein complex plays a key role in recognizing DNA damage throughout the genome for mammalian nucleotide excision repair (NER). Ultraviolet light (UV)-damaged DNA binding protein (UV-DDB) is another complex that appears to be involved in the recognition of NER-inducing damage, although the precise role it plays and its relationship to XPC remain to be elucidated. Here we show that XPC undergoes reversible ubiquitylation upon UV irradiation of cells and that this depends on the presence of functional UV-DDB activity. XPC and UV-DDB were demonstrated to interact physically, and both are polyubiquitylated by the recombinant UV-DDB-ubiquitin ligase complex. The polyubiquitylation altered the DNA binding properties of XPC and UV-DDB and appeared to be required for cell-free NER of UV-induced (6-4) photoproducts specifically when UV-DDB was bound to the lesion. Our results strongly suggest that ubiquitylation plays a critical role in the transfer of the UV-induced lesion from UV-DDB to XPC. PMID- 15882622 TI - Developmental commitment in a bacterium. AB - We investigated developmental commitment during sporulation in Bacillus subtilis. Sporulation is initiated by nutrient limitation and involves division of the developing cell into two progeny, the forespore and the mother cell, with different fates. Differentiation becomes irreversible following division when neither the forespore nor the mother cell can resume growth when provided with nutrients. We show that commitment is governed by the transcription factors sigma(F) and sigma(E), which are activated in the forespore and the mother cell, respectively. We further show that commitment involves spoIIQ, which is under the control of sigma(F), and spoIIP, which is under the control of both sigma(F) and sigma(E). In the presence of nutrients, the forespore can exhibit rodlike, longitudinal growth when SpoIIQ and SpoIIP are absent, whereas the mother cell can do so when SpoIIP alone is absent. Thus, developmental commitment of this single-celled organism, like that of the cells of complex, multicellular organisms, ensures that differentiation is maintained despite changes in the extracellular milieu. PMID- 15882623 TI - A new paradigm for translational control: inhibition via 5'-3' mRNA tethering by Bicoid and the eIF4E cognate 4EHP. AB - Translational control is a key genetic regulatory mechanism implicated in regulation of cell and organismal growth and early embryonic development. Initiation at the mRNA 5' cap structure recognition step is frequently targeted by translational control mechanisms. In the Drosophila embryo, cap-dependent translation of the uniformly distributed caudal (cad) mRNA is inhibited in the anterior by Bicoid (Bcd) to create an asymmetric distribution of Cad protein. Here, we show that d4EHP, an eIF4E-related cap binding protein, specifically interacts with Bcd to suppress cad translation. Translational inhibition depends on the Bcd binding region (BBR) present in the cad 3' untranslated region. Thus, simultaneous interactions of d4EHP with the cap structure and of Bcd with BBR renders cad mRNA translationally inactive. This example of cap-dependent translational control that is not mediated by canonical eIF4E defines a new paradigm for translational inhibition involving tethering of the mRNA 5' and 3' ends. PMID- 15882624 TI - Polycomb group protein ezh2 controls actin polymerization and cell signaling. AB - Polycomb group protein Ezh2, one of the key regulators of development in organisms from flies to mice, exerts its epigenetic function through regulation of histone methylation. Here, we report the existence of the cytosolic Ezh2 containing methyltransferase complex and tie the function of this complex to regulation of actin polymerization in various cell types. Genetic evidence supports the essential role of cytosolic Ezh2 in actin polymerization-dependent processes such as antigen receptor signaling in T cells and PDGF-induced dorsal circular ruffle formation in fibroblasts. Revealed function of Ezh2 points to a broader usage of lysine methylation in regulation of both nuclear and extra nuclear signaling processes. PMID- 15882625 TI - Modulation of receptor recycling and degradation by the endosomal kinesin KIF16B. AB - Different classes of endosomes exhibit a characteristic intracellular steady state distribution governed by interactions with the cytoskeleton. We found a kinesin-3, KIF16B, that transports early endosomes to the plus end of microtubules in a process regulated by the small GTPase Rab5 and its effector, the phosphatidylinositol-3-OH kinase hVPS34. In vivo, KIF16B overexpression relocated early endosomes to the cell periphery and inhibited transport to the degradative pathway. Conversely, expression of dominant-negative mutants or ablation of KIF16B by RNAi caused the clustering of early endosomes to the perinuclear region, delayed receptor recycling to the plasma membrane, and accelerated degradation. These results suggest that KIF16B, by regulating the plus end motility of early endosomes, modulates the intracellular localization of early endosomes and the balance between receptor recycling and degradation. We propose that this mechanism could have important implications for signaling. PMID- 15882626 TI - Nuclear movement regulated by Cdc42, MRCK, myosin, and actin flow establishes MTOC polarization in migrating cells. AB - The microtubule-organizing center (MTOC) is reoriented between the nucleus and the leading edge in many migrating cells and contributes to directional migration. Models suggest that the MTOC is moved to its position during reorientation. By direct imaging of wound-edge fibroblasts after triggering MTOC reorientation with soluble factors, we found instead that the nucleus moved away from the leading edge to reorient the MTOC, while the MTOC remained stationary. Rearward nuclear movement was coupled with actin retrograde flow and was regulated by a pathway involving Cdc42, MRCK, myosin, and actin. Nuclear movement was unaffected by the inhibition of dynein, Par6, or PKCzeta, yet these components were essential for MTOC reorientation, as they maintained the MTOC at the cell centroid. These results show that nuclear repositioning is an initial polarizing event in migrating cells and that the positions of the nucleus and the MTOC are established by separate regulatory pathways. PMID- 15882627 TI - Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues. AB - The POU-domain transcription factor Oct-4 is normally expressed in pluripotent cells of the mammalian embryo. In addition, germ-cell tumors and a few somatic tumors show detectable expression of Oct-4. While Oct-4's role during preimplantation development is to maintain embryonic cells in a pluripotent state, little is known about its potential oncogenic properties. Here we investigate the effect of ectopic Oct-4 expression on somatic tissues of adult mice using a doxycycline-dependent expression system. Activation of Oct-4 results in dysplastic growths in epithelial tissues that are dependent on continuous Oct 4 expression. Dysplastic lesions show an expansion of progenitor cells and increased beta-catenin transcriptional activity. In the intestine, Oct-4 expression causes dysplasia by inhibiting cellular differentiation in a manner similar to that in embryonic cells. These data show that certain adult progenitors remain competent to interpret key embryonic signals and support the notion that progenitor cells are a driving force in tumorigenesis. PMID- 15882628 TI - Mosaic analysis with double markers in mice. AB - We describe a method termed MADM (mosaic analysis with double markers) in mice that allows simultaneous labeling and gene knockout in clones of somatic cells or isolated single cells in vivo. Two reciprocally chimeric genes, each containing the N terminus of one marker and the C terminus of the other marker interrupted by a loxP-containing intron, are knocked in at identical locations on homologous chromosomes. Functional expression of markers requires Cre-mediated interchromosomal recombination. MADM reveals that interchromosomal recombination can be induced efficiently in vivo in both mitotic and postmitotic cells in all tissues examined. It can be used to create conditional knockouts in small populations of labeled cells, to determine cell lineage, and to trace neuronal connections. To illustrate the utility of MADM, we show that cerebellar granule cell progenitors are fated at an early stage to produce granule cells with axonal projections limited to specific sublayers of the cerebellar cortex. PMID- 15882629 TI - Taking two TRAILS. AB - The TNF-related apoptosis-inducing ligand (TRAIL) has well-described anti inflammatory effects in models of autoimmune disease, including experimental autoimmune encephalomyelitis (EAE). In this issue of Neuron, Aktas, Smorodchenko, and colleagues present evidence that TRAIL exerts anti-inflammatory effects, but also induces neuronal apoptosis, in EAE. This report poses the therapeutic challenge of facilitating TRAIL expression in the periphery while inhibiting TRAIL in the CNS. PMID- 15882630 TI - Retinal coding of visual scenes -- repetitive and redundant too? AB - Visual information reaches the brain by way of a fine cable, the optic nerve. The million or so axons in the optic nerve represent an information bottleneck in the visual pathway-where the fewest number of neurons convey the visual scene. It has long been thought that to make the most of the optic nerve's limited capacity the retina may encode visual information in an optimally efficient manner. In this issue of Neuron, Puchalla et al. report a test of this hypothesis using multielectrode recordings from retinal ganglion cells stimulated with movies of natural scenes. The authors find substantial redundancy in the retinal code and estimate that there is an approximately 10-fold overrepresentation of visual information. PMID- 15882631 TI - Cortical development: new concepts. PMID- 15882632 TI - Stem cells: from epigenetics to microRNAs. AB - The complexity and cellular diversity of the adult brain arises from the proliferation and differentiation of a small number of stem cells. The intrinsic state of stem cells depends on their spatial and temporal history and affects their responsiveness to extrinsic signals from the microenvironment. Stem cell self-renewal and differentiation along neuronal and glial lineages are defined by the dynamic interplay between transcription, epigenetic control, and posttranscriptional regulators, including microRNAs, whose key role in stem cell biology is just emerging. PMID- 15882633 TI - Radial glial cells defined and major intermediates between embryonic stem cells and CNS neurons. AB - Radial glial cells have been identified as a major source of neurons during development. Here, we review the evidence for the distinct "glial" nature of radial glial cells and contrast these cells with their progenitors, the neuroepithelial cells. Recent results also suggest that not only during neurogenesis in vivo, but also during the differentiation of cultured embryonic stem cells toward neurons, progenitors with clear glial antigenic characteristics act as cellular intermediates. PMID- 15882634 TI - Cortical neuron specification: it has its time and place. AB - Cortical neurogenesis is a highly stereotyped process in which progenitor cells generate neurons destined for specific cortical layers depending on the timing of cell cycle exit. Previous work has shown that during corticogenesis, progenitors become progressively restricted in their developmental potential. Recent work has uncovered some of the intrinsic mechanisms that underlie this fate restriction. In addition to timing, new studies suggest that the location of cell cycle exit in the cortical germinal zone may also contribute to cortical neuron specification. PMID- 15882635 TI - Developmental mechanisms underlying the generation of cortical interneuron diversity. AB - GABAergic interneurons are critical components of cortical circuits. However, understanding their function has become extremely challenging because they constitute one of the most diverse groups of cells in the central nervous system. Indeed, cortical GABAergic interneurons are heterogeneous in so many different ways--morphology, molecular profiling, electrical properties--that even attempts to discern what parameters should be used to identify cortical interneuron subtypes have failed to generate broad consensus among experts in the field. The extent to which cortical interneuron diversity emerges during development is largely unknown, but it is likely that insights on how this process takes place may help us understand their role as integrative and synchronizing elements in cortical function. Here, we review recent data on how the large variety of distinct classes of cortical interneurons may arise during development. PMID- 15882636 TI - Nucleokinesis in neuronal migration. AB - Neuronal migration is a critical phase of nervous system development and can be divided into two distinct phases: extension of the leading process and movement of the cell body and nucleus (nucleokinesis). Nucleokinesis appears to require many of the same cytoskeletal and signaling molecules used in cell mitosis. Converging studies suggest it requires cytoplasmic dynein, cell polarity genes, and microtubule-associated proteins that coordinate microtubule remodeling. These coordinate first the positioning of the centrosome (microtubule organizing center) in the leading process in front of the nucleus and then the movement of the nucleus towards the centrosome. The positioning of the centrosome and the dynamic regulation that couples and uncouples the nucleus underlies directed migration of neurons. PMID- 15882637 TI - The cells of cajal-retzius: still a mystery one century after. AB - Cajal-Retzius (CR) cells are an enigmatic class of neurons located at the surface of the cerebral cortex, playing a major role in cortical development. In this review, we discuss several distinct features of these neurons and the mechanisms by which they regulate cortical development. Many CR cells likely have extracortical origin and undergo cell death during development. Recent genetic studies report unique patterns of gene expression in CR cells, which may help to explain the developmental processes in which they participate. Moreover, a number of studies indicate that CR cells, and their secreted gene product, reelin, are involved in neuronal migration by acting on two key partners, migrating neurons and radial glial cells. Emerging data show that these neurons are a critical part of an early and complex network of neural activity in layer I, supporting the notion that CR cells modulate cortical maturation. Given these key and complex developmental properties, it is therefore conceivable for CR cells to be implicated in the pathogenesis of a variety of neurological disorders. PMID- 15882638 TI - Genetic mechanisms specifying cortical connectivity: let's make some projections together. AB - Great neuroanatomists of the twentieth century recognized that the cerebral cortex of mammals is the single most complex structure of the central nervous system both in terms of neuronal diversity and connectivity. Understanding the cellular and molecular mechanisms specifying the afferent and efferent connectivity in the neocortex may seem like a daunting task. However, recent technical advances have greatly improved our ability to (1) profile gene expression of neuronal populations isolated based on their connectional properties, (2) manipulate gene expression in specific neuronal populations, and (3) visualize their axonal projections in vivo. These new tools are revolutionizing our ability to identify the molecular mechanisms patterning afferent and efferent cortical projections. PMID- 15882639 TI - Calcium signaling and the control of dendritic development. AB - Dendrites serve a critical role in neuronal information processing as sites of synaptic integration. The morphological diversity of dendritic architecture reflects specialized strategies that neurons have evolved to detect and process incoming information. Recent observations suggest that calcium signals exert an important influence on neuronal morphology by regulating the growth and branching of dendrites and the formation of dendritic spines. Calcium signals appear to influence branch dynamics by affecting the cytoskeleton near the site of calcium entry, whereas calcium-dependent dendritic growth involves activation of a transcriptional program. PMID- 15882640 TI - Developmental neurobiology and clinical disorders: lost in translation? AB - Advances in defining mechanisms of cortical development have been paralleled in recent years by an intense interest in translating these findings into greater insight of both childhood- and adult-onset cognitive and mental health disorders of developmental etiology. Successful integration of basic and clinical findings have been applied to monogenic disorders. The greater challenge lies in studying cortical development in the context of gene x environment interactions, which underlie the pathogenesis of the most common neurodevelopmental disorders. This can occur through an improved delineation of pathophysiological characteristics unique to specific complex disorders and the application of this information to the refinement of the most relevant model systems. PMID- 15882641 TI - Recoverin improves rod-mediated vision by enhancing signal transmission in the mouse retina. AB - Vision in dim light requires that photons absorbed by rod photoreceptors evoke signals that reliably propagate through the retina. We investigated how a perturbation in rod physiology affects propagation of those signals in the retina and ultimately visual sensitivity. Recoverin is a protein in rods that prolongs phototransduction and enhances visual sensitivity. It is not present in neurons postsynaptic to rods, yet we found that light-evoked responses of rod bipolar and ganglion cells were shortened when measured in recoverin-deficient retinas. Unexpectedly, the effect of recoverin on postsynaptic signals could not be explained by its effect on phototransduction. Instead, it is an effect of recoverin downstream of phototransduction in rods that prolongs signal transmission and enhances visual sensitivity. An important implication of our findings is that the recovery phase of the rod photoresponse does not contribute significantly to visual sensitivity near absolute threshold. PMID- 15882642 TI - Neuronal damage in autoimmune neuroinflammation mediated by the death ligand TRAIL. AB - Here, we provide evidence for a detrimental role of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in neural death in T cell-induced experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). Clinical severity and neuronal apoptosis in brainstem motor areas were substantially reduced upon brain-specific blockade of TRAIL after induction of EAE through adoptive transfer of encephalitogenic T cells. Furthermore, TRAIL deficient myelin-specific lymphocytes showed reduced encephalitogenicity when transferred to wild-type mice. Conversely, intracerebral delivery of TRAIL to animals with EAE increased clinical deficits, while naive mice were not susceptible to TRAIL. Using organotypic slice cultures as a model for living brain tissue, we found that neurons were susceptible to TRAIL-mediated injury induced by encephalitogenic T cells. Thus, in addition to its known immunoregulatory effects, the death ligand TRAIL contributes to neural damage in the inflamed brain. PMID- 15882643 TI - Pathological cell-cell interactions elicited by a neuropathogenic form of mutant Huntingtin contribute to cortical pathogenesis in HD mice. AB - Expanded polyglutamine (polyQ) proteins in Huntington's disease (HD) as well as other polyQ disorders are known to elicit a variety of intracellular toxicities, but it remains unclear whether polyQ proteins can elicit pathological cell-cell interactions which are critical to disease pathogenesis. To test this possibility, we have created conditional HD mice expressing a neuropathogenic form of mutant huntingtin (mhtt-exon1) in discrete neuronal populations. We show that mhtt aggregation is a cell-autonomous process. However, progressive motor deficits and cortical neuropathology are only observed when mhtt expression is in multiple neuronal types, including cortical interneurons, but not when mhtt expression is restricted to cortical pyramidal neurons. We further demonstrate an early deficit in cortical inhibition, suggesting that pathological interactions between interneurons and pyramidal neurons may contribute to the cortical manifestation of HD. Our study provides genetic evidence that pathological cell cell interactions elicited by neuropathogenic forms of mhtt can critically contribute to cortical pathogenesis in a HD mouse model. PMID- 15882644 TI - The molecular basis of odor coding in the Drosophila larva. AB - We have analyzed the molecular basis of odor coding in the Drosophila larva. A subset of Or genes is found to be expressed in larval olfactory receptor neurons (ORNs). Using an in vivo expression system and electrophysiology, we demonstrate that these genes encode functional odor receptors and determine their response spectra with 27 odors. The receptors vary in their breadth of tuning, exhibit both excitation and inhibition, and show different onset and termination kinetics. An individual receptor appears to transmit signals via a single ORN to a single glomerulus in the larval antennal lobe. We provide a spatial map of odor information in the larval brain and find that ORNs with related functional specificity map to related spatial positions. The results show how one family of receptors underlies odor coding in two markedly different olfactory systems; they also provide a molecular mechanism to explain longstanding observations of larval odor discrimination. PMID- 15882645 TI - Connecting the navigational clock to sun compass input in monarch butterfly brain. AB - Migratory monarch butterflies (Danaus plexippus) use a time-compensated sun compass to navigate to their overwintering grounds in Mexico. Although polarized light is one of the celestial cues used for orientation, the spectral content (color) of that light has not been fully explored. We cloned the cDNAs of three visual pigment-encoding opsins (ultraviolet [UV], blue, and long wavelength) and found that all three are expressed uniformly in main retina. The photoreceptors of the polarization-specialized dorsal rim area, on the other hand, are monochromatic for the UV opsin. Behavioral studies support the importance of polarized UV light for flight orientation. Next, we used clock protein expression patterns to identify the location of a circadian clock in the dorsolateral protocerebrum of butterfly brain. To provide a link between the clock and the sun compass, we identified a CRYPTOCHROME-staining neural pathway that likely connects the circadian clock to polarized light input entering brain. PMID- 15882646 TI - Long-term plasticity mediated by mGluR1 at a retinal reciprocal synapse. AB - The flow of information across the retina is controlled by reciprocal synapses between bipolar cell terminals and amacrine cells. However, the synaptic delays and properties of plasticity at these synapses are not known. Here we report that glutamate release from goldfish Mb-type bipolar cell terminals can trigger fast (delay of 2-3 ms) and transient GABA(A) IPSCs and a much slower and more sustained GABA(C) feedback. Synaptically released glutamate activated mGluR1 receptors on amacrine cells and, depending on the strength of presynaptic activity, potentiated subsequent feedback. This poststimulus enhancement of GABAergic feedback lasted for up to 10 min. This form of mGluR1-mediated long term synaptic plasticity may provide retinal reciprocal synapses with adaptive capabilities. PMID- 15882647 TI - Fast odor learning improves reliability of odor responses in the locust antennal lobe. AB - Recordings in the locust antennal lobe (AL) reveal activity-dependent, stimulus specific changes in projection neuron (PN) and local neuron response patterns over repeated odor trials. During the first few trials, PN response intensity decreases, while spike time precision increases, and coherent oscillations, absent at first, quickly emerge. We examined this "fast odor learning" with a realistic computational model of the AL. Activity-dependent facilitation of AL inhibitory synapses was sufficient to simulate physiological recordings of fast learning. In addition, in experiments with noisy inputs, a network including synaptic facilitation of both inhibition and excitation responded with reliable spatiotemporal patterns from trial to trial despite the noise. A network lacking fast plasticity, however, responded with patterns that varied across trials, reflecting the input variability. Thus, our study suggests that fast olfactory learning results from stimulus-specific, activity-dependent synaptic facilitation and may improve the signal-to-noise ratio for repeatedly encountered odor stimuli. PMID- 15882648 TI - Redundancy in the population code of the retina. AB - We have explored the manner in which the population of retinal ganglion cells collectively represent the visual world. Ganglion cells in the salamander were recorded simultaneously with a multielectrode array during stimulation with both artificial and natural visual stimuli, and the mutual information that single cells and pairs of cells conveyed about the stimulus was estimated. We found significant redundancy between cells spaced as far as 500 mum apart. When we used standard methods for defining functional types, only ON-type and OFF-type cells emerged as truly independent information channels. Although the average redundancy between nearby cell pairs was moderate, each ganglion cell shared information with many neighbors, so that visual information was represented approximately 10-fold within the ganglion cell population. This high degree of retinal redundancy suggests that design principles beyond coding efficiency may be important at the population level. PMID- 15882649 TI - Implicit associative learning engages the hippocampus and interacts with explicit associative learning. AB - The hippocampus is crucial for conscious, explicit memory, but whether it is also involved in nonconscious, implicit memory is uncertain. We investigated with functional magnetic resonance imaging whether implicit learning engages the hippocampus and interacts with subsequent explicit learning. The presentation of subliminal faces-written profession pairs for implicit learning was followed by the explicit learning of supraliminal pairs composed of the same faces combined with written professions semantically incongruous to those presented subliminally (experiment 1), semantically congruous professions (experiment 2), or identical professions (experiment 3). We found that implicit face-profession learning interacted with explicit face-profession learning in all experiments, impairing the explicit retrieval of the associations. Hippocampal activity increased during the subliminal presentation of face-profession pairs versus face-nonword pairs and correlated with the later impairment of explicit retrieval. These findings suggest that implicit semantic associative learning engages the hippocampus and influences explicit memory. PMID- 15882650 TI - The place of Amborella within the radiation of angiosperms. PMID- 15882651 TI - Chloroplast genome phylogenetics: why we need independent approaches to plant molecular evolution. AB - The traditional approach to plant molecular phylogenetics involves amplifying, sequencing and analyzing one or a few genes from many species and is conducive to broad taxon sampling. An independent approach involves chloroplast genome sequencing, providing much larger amounts of data per taxon but for a smaller number of species. In principle, the two strategies can inform each other but in practice their results sometimes conflict for reasons that are currently debated. An Opinion article published in the October 2004 issue of Trends in Plant Science cautioned against the pursuit of genome-based phylogenies. Here, we provide a different perspective on issues at the heart of the current debate and defend the use of chloroplast genome phylogenetics for crucial species because it provides an independent test of hypotheses generated by the traditional approach. PMID- 15882652 TI - What genes make a tree a tree? AB - Woody growth is evolutionarily ancient, yet has been gained and lost multiple times in plant evolution and is readily enhanced or minimized in eudicot speciation. New molecular genetic and genomic studies in Populus and Arabidopsis that are defining the genes responsible for cambium function and woody growth suggest that the genes regulating woody growth are not unique to woody plants. Surprisingly, key genetic mechanisms originally characterized as regulating the meristematic cells of the shoot apical meristem are also expressed in the vascular cambium during woody growth. This has important implications for the development of Populus as a model species and illustrates why forest trees constitute a contrived group of plants that have more in common with herbaceous relatives than we foresters like to admit. PMID- 15882653 TI - The future of C4 research--maize, Flaveria or Cleome? AB - C4 photosynthesis has evolved multiple times among the angiosperms: the spatial rearrangement of the photosynthetic apparatus, combined with alterations to the leaf structure, allows CO2 to be concentrated around Rubisco. Higher CO2 concentrations at Rubisco decrease the rate of oxygenation and therefore reduce the amount of energy lost through photorespiration. C4 plants are particularly prevalent in tropical and subtropical regions because they can sustain higher rates of net photosynthesis; they also represent some of our most productive crops. To date, most progress in identifying genes crucial for C4 photosynthesis has been made using maize and Flaveria. We propose that Cleome, the most closely related genus containing C4 species to the C3 model Arabidopsis, be used together with Arabidopsis resources to accelerate our progress in elucidating the genetic basis of C4 photosynthesis. PMID- 15882654 TI - Lotus japonicus: legume research in the fast lane. AB - Legumes are of immense importance to humanity and a key to sustainable agriculture. Two model species, Lotus japonicus and Medicago truncatula, are the focus of genome sequencing and functional genomics programmes, but most researchers focus exclusively on one or the other. In spite of this, legume researchers now have a unique opportunity to integrate work on these and other legume species, including soybean, common bean and pea to create a platform for comparative genomics second to none of any other plant family. The question is: do we have the scientific fortitude and political will to achieve this? PMID- 15882655 TI - Insertional mutagenesis: a Swiss Army knife for functional genomics of Medicago truncatula. AB - Legumes are second only to grasses in worldwide economic importance, and understanding their molecular genetics is vital to the breeding of important grain and forage legumes. Over the past decade, Medicago truncatula has been selected as a model plant in which to study biological processes that are unique and pertinent to legumes, and that cannot easily be studied in Arabidopsis. Here, we discuss the most common tools for introducing and analyzing genetic mutations in M. truncatula. Because transformation and regeneration are still bottlenecks in studying a legume species, large-scale insertional mutagenesis poses a major challenge in M. truncatula. We discuss the tobacco retrotransposon Tnt1 as a viable and attractive option for introducing multiple independent insertions per plant for saturation mutagenesis. PMID- 15882656 TI - Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. AB - For more than a century, the biosynthesis of flavonoid pigments has been a favorite of scientists to study a wide variety of biological processes, such as inheritance and transposition, and has become one of the best-studied pathways in nature. The analysis of pigmentation continues to provide insights into new areas, such as the channeling and intracellular transport of metabolites, regulation of gene expression and RNA interference. Moreover, because pigmentation is studied in a variety of species, it provides unique molecular insights into the evolution of biochemical pathways and regulatory networks. PMID- 15882657 TI - Use of Petunia to unravel plant meristem functioning. AB - In the past decade, enormous progress has been made in our understanding of the molecular and genetic control of meristem growth, maintenance and differentiation into plant organs. Several model plants have contributed to our current knowledge of meristem function. Research using Petunia has had a substantial share in this progress. Integration of information obtained from this species gives clues about the common and diverged pathways underlying the formation and functioning of plant meristems. PMID- 15882658 TI - A model system for comparative research: Petunia. AB - Research today aims to analyse the development of plant processes over evolutionary time. To obtain a representative view, a range of plant species covering at least the crucial nodes in phylogeny must be selected for an in depth analysis. Here we present Petunia as one of the available systems: as a representative of the Solanaceae it has the advantages of good culture conditions and the availability of a range of materials, techniques and strategies that can be used to research an interesting and diverse set of questions. PMID- 15882659 TI - Brugada syndrome. AB - The Brugada syndrome is an autosomal dominant disease with incomplete penetrance that may cause syncope and sudden cardiac death in young individuals with a normal heart. It is characterized by an electrocardiographic pattern of complete or incomplete right bundle branch block and ST segment elevation in leads V1-V3. One of the genes linked to this syndrome is SCN5A, the gene encoding for the cardiac sodium channel. Mutations in SCN5A cause a functional reduction in the availability of cardiac sodium current in Brugada syndrome. However, only 20-25% of patients affected by this syndrome have mutations on this gene. A novel gene locus on chromosome 3, distinct from SCN5A, has been identified recently. The relative male preponderance of the phenotype, despite equal inheritance of the gene in males and females, has led to the speculation of a role for testosterone in the phenotype. The disease could manifest at first time as cardiac arrest without any previous symptom, and the electrocardiographic pattern could be intermittent, requiring a pharmacological challenge with Class I antiarrhythmic drugs to unmask ST elevation. Several conditions producing Brugada-like electrocardiographic patterns should be borne in mind and excluded while making a diagnosis of the Brugada syndrome. The management is difficult as pharmacological agents are not universally effective. The mode of treatment recommended by the majority of cardiac electrophysiologists is the implantation of a cardioverter defibrillator. Symptomatic patients with inducible ventricular arrhythmias and a positive family history should be considered for prophylactic implantation of a cardioverter defibrillator. PMID- 15882660 TI - Age- and sex-related utilisation of cardiac procedures and interventions: a multicentric study in Italy. AB - BACKGROUND: Cardiovascular diseases represent the leading cause of death in Italy and one of the most frequent cause of disability in the elderly. The aim of the study was to investigate the influence of age and sex of patient on the utilisation of cardiac procedure and interventions in Italy. METHODS: Retrospective analysis of 2805 patients' medical case notes in cardiology, internal medicine and elderly clinics in six Italian Regions during the period 1996-1997, considering coronary catheterisation (CC), percutaneous transluminal coronary angioplasty (PCTA), coronary artery bypass grafting (CABG), thrombolysis (THR) and permanent pacemaker (PPM) implantation. RESULTS: Older patients (more than 75 years old) were less likely to undergo CC (OR=0.062). Trends for age and sex did not achieve significance for CAGB (OR=0.815 for older patient). Age is a strong predictor of receiving PTCA, with the oldest group of patients being discriminated (OR=0.093 for people older than 75 years), and the same trend was observed for THR (OR=0.264 for patients older than 75 years). For PPM, older patient has a higher likelihood of receiving this type of intervention (OR=3.45 for 65-74 years, and OR=7.77 for patients older than 75 years). As far as gender of patients is concerned, statistically significant differences for all cardiac procedures or interventions considered were not found. CONCLUSIONS: Clinical management of older patients with cardiac disease in Italy may be different from that of younger patients. One possible explanation would be that these patients are being discriminated against mainly because of their age. PMID- 15882661 TI - Acute myocardial infarction in young patients: nutritional status and biochemical factors. AB - PURPOSE: The aim of this study was to establish whether nutritional status and biochemical factors, C-reactive protein (CRP), serum amyloid A (SAA) protein, serum iron (Fe) and fibrinogen at admission were different in patients with acute myocardial infarction (AMI) at a young age (<40 years) vs. those with AMI at an older age (>60 years). We also investigated whether during the stay in the hospital, the increase in acute-phase reactants was different in young vs. older subjects, and if dyslipidemic aspects were different between the two groups. METHODS: The study population consisted of 40 patients, all males with a mean age of 36.7+/-1.16 years, admitted to our facility with AMI. The control group included 40 patients, all males, mean age of 66.3+/-4.24 years, with AMI. CRP, SAA, Fe and fibrinogen were determined at admission to the hospital and daily for 7 days in the two groups of patients. RESULTS: In young patients the median value of the highest levels were 6.2 mg/l (range 0.7-27.30) for CRP, 13.22 mg/l (range 0.7-130) for SAA, 420 mg/dl (range 76-840) for fibrinogen and 49.1 gamma/ml (range 14-102) for Fe levels. In the older patients, the median value of the highest levels were 5.9 mg/l (range 0.6-28.30) for CRP, 12.12 mg/l (range 0.9 280) for SAA, 480 mg/dl (range 60-780) for fibrinogen and 47.1 gamma/ml (range 12 94) for Fe levels. CONCLUSIONS: In the present study, acute-phase reactants were quantitatively similar in young and old patients. On the contrary, nutritional status, homocysteine, LDL and triglycerides are significantly higher in young patients than in old patients. PMID- 15882662 TI - AMPD1 (C34T) polymorphism and clinical outcomes in patients undergoing myocardial revascularization. AB - BACKGROUND: C34T variant of adenosine monophosphate deaminase 1 (AMPD1) gene has been associated with a prolonged survival in heart failure and coronary artery disease, hypothetically linked to an enhanced production of adenosine. DESIGN: Since adenosine administration is a promising approach for the prevention of the ischemia-reperfusion in myocardial revascularization, the aim of this study was to investigate whether the AMPD1 (-) allele is associated with a favorable prognosis after coronary revascularization. In addition, we assessed the association between AMPD1 polymorphism and plasma adenosine levels. METHODS: We investigated a total of 161 patients receiving coronary revascularization (70 percutaneous transluminal coronary angioplasty and 91 coronary artery bypass graft). They were investigated for a composite endpoint including recurrent angina, non-fatal MI, target vessel revascularization, heart failure and cardiac death. Plasma adenosine was also measured by high-performance liquid chromatography methods on a subset of 25 patients. RESULTS: During the follow-up period (7.0+/-0.3 months), the overall combined endpoint accounted for 17 events (10 cardiac-related deaths, 6 revascularization procedures and 1 congestive heart failure). The composite endpoint was 9.8% for AMPD1 (-) allele carriers vs. 11.5% for non-carriers (log-rank statistic, p=n.s.). In the logistic analysis only low (95th percentile for age and sex. We divided the NF1 group into two subgroups: group A, patients with 24-h SBP and DBP 95th percentile for age and sex. RESULTS: Group B presented a thicker end-diastolic interventricular septum (p<0.0001), posterior wall (p=0.02), LVMI (p<0.001) and relative wall thickness (p<0.03) than group A and controls. Left atrial dimension in group B was also significantly larger. Examination by standard Doppler showed a deceleration and isovolumic relaxation time significantly prolonged in group B. DTI parameters were significantly higher in NF1 patients than controls. In group B, myocardial early diastolic (E(m)) and systolic (S(m)) velocities were significantly lower than group A. Myocardial early/late diastolic ratio (E(m)/A(m)) in NF1 patients was lower than controls and 19% of group A and 20% of group B showed an E(m)/A(m) ratio <1. No healthy subjects presented an E(m)/A(m) ratio <1. CONCLUSIONS: We demonstrated early cardiac morphologic and functional changes in young NF1 patients with hypertension. Because DTI directly studies cardiac muscle, it can detect changes induced by hypertension as well as those independent of blood pressure. PMID- 15882672 TI - Platelet-monocyte aggregates predict troponin rise after percutaneous coronary intervention and are inhibited by Abciximab. AB - BACKGROUND: Platelet-monocyte aggregates and other markers of platelet activation were investigated before and after percutaneous coronary intervention (PCI) with abciximab therapy. The study sought to assess the relationship between the level of platelet-monocyte aggregation and increases in cardiac troponin I post coronary intervention. METHODS: Blood samples were collected from 40 patients before PCI and 10 min after abciximab administration. These were tested for platelet activation markers by flow cytometry. Cardiac troponin I levels were assayed at baseline and at 24 h post PCI. RESULTS: Compared to healthy controls, patients with coronary artery disease had elevated markers of platelet activation including platelet-monocyte aggregates, P-selectin and PAC-1 (a marker specific for activated glycoprotein IIb/IIIa) prior to PCI. Increased levels of platelet monocyte aggregates before PCI were associated with increased expression of P selectin on the platelet surface. Abciximab therapy reduced platelet-monocyte aggregate levels but had no effect on P-selectin expression. The high levels of expression of activated glycoprotein IIb/IIIa (PAC-1) on platelets prior to PCI was reduced with abciximab therapy. Patients with higher levels of platelet monocyte aggregates prior to PCI were more likely to develop an elevation of cardiac troponin I during the 24 h after PCI. CONCLUSIONS: Increased levels of platelet-monocyte aggregates may predict patients at risk for troponin elevation following PCI and identify those most likely to benefit from abciximab. PMID- 15882673 TI - The influence of growth hormone therapy on ultrasound myocardial tissue characterization in patients with childhood onset GH deficiency. AB - BACKGROUND: In growth hormone deficiency (GHD), a reduction in left ventricular mass (LV-mass) and impairment of systolic function has been shown. In this study, we investigated the effects of 12 months of GH replacement therapy on cardiac structure and functional indices measured by echocardiographic techniques in adult patients with childhood onset GH deficiency. METHODS: Sixteen patients (age 42.3+/-13.1 years, 10 males) were investigated before and after 12 months of GH treatment at a dose of 0.02 IU/kg/day (7 microg/kg/day). Echocardiography was performed including the ultrasound myocardial tissue characterization technique. We measured two parameters of the ultrasonic tissue characterization with integrated backscatter: the magnitude of the cardiac-cycle-dependent variation in integrated backscatter signals (CV-IBS) and the mean value of integrated backscatter signals calibrated by the pericardium (cal-IBS). RESULTS: Left ventricular diameter and wall thickness did not change after GH treatment, although systolic increase in interventricular septum thickness (IVS%) and systolic increase in posterior wall thickness (PWT%) increased significantly (IVS% 52.2+/-31.9% vs. 67.3+/-30.4% and PWT% 48.7+/-20.2% vs. 58.0+/-17.7%, p<0.01 and p<0.01, respectively). Ejection fraction increased from 56.2+/-7.2% to 63.2+/-6.1% (p<0.01). LV-mass index did not change after GH treatment (78.4+/ 22.1 vs. 81.9+/-21.1 g/m(2)). CV-IBS increased significantly after GH treatment (p<0.05), in both the interventricular septum and the left ventricular posterior wall (4.7+/-1.5 vs. 5.8+/-1.9 dB for the interventricular septum, 4.9+/-1.8 vs. 6.5+/-2.4 dB for the left ventricular posterior wall, p<0.05 and p<0.05, respectively). Cal-IBS also increased significantly after GH treatment (-23.5+/ 4.1 vs.-21.8+/-4.2 dB for the interventricular septum, -23.0+/-4.4 vs. -21.8+/ 4.3 dB for the left ventricular posterior wall, p<0.01 and p<0.05, respectively). CONCLUSION: Twelve months GH treatment in adults with childhood onset GHD resulted in improvement of cardiac contractile performance. Observed changes in cal-IBS and CV-IBS suggest that GH treatment in this patient group can lead to a further somatic maturation of the heart, probably not accomplished previously. PMID- 15882674 TI - The human FGF2 level is influenced by genetic predisposition. AB - BACKGROUND: The fibroblast growth factor 2 (FGF2) is involved in various processes possibly leading to the development of complex diseases such as atherosclerosis. In recent studies, its cardioprotective properties, due to its ability to stimulate the proliferation of collateral vessels, could be shown. STUDY DESIGN: In this clinical study, the relation between clinical risk markers, a genomic variant of FGF2, namely the c.223C>T polymorphism, and the in vivo FGF2 expression was evaluated. Therefore, 198 clinically well-characterized probands, all of Caucasian origin, were included. The FGF2 mRNA level was determined in monocytes by competitive RT-PCR, whereas the plasma level of circulating FGF2 protein was analysed by ELISA. By considering the angiographically proven stenotic state of the patient, a significant increase in FGF2 mRNA, but not in protein level, could be shown for patients with significant stenosis. Apart from this, no influence on FGF2 expression was found in the case of all of the clinical and biochemical markers investigated. However, in the case of the c.223C>T polymorphism, a significant increase in the individual FGF2 mRNA and protein level in CC-carriers was shown. In multivariate analysis, this relation was independent of all other risk markers investigated. CONCLUSIONS: Our results suggest that an increase in FGF2 mRNA expression, related to coronary atherosclerosis, may be necessary for the maintenance of the individual FGF2 plasma level. Since the individual FGF2 mRNA and protein level are, to a large extent, triggered off by genetic background, the FGF2 expression cannot be referred to as an independent clinical marker for CAD. PMID- 15882675 TI - Effect of posture on heart rate variability spectral measures in children and young adults with heart disease. AB - BACKGROUND: Reduction of heart rate variability as a consequence of heart disease and postural change has been well documented. However, the data on the effect of postural change in pediatric patients are incomplete and the effect is not fully understood. The aim of the study was to investigate effect of postural change on heart rate variability in relation to the extent of severity of heart disease. METHODS: The dependence of heart rate variability on posture in 41 children and young adults (8-20 years) with heart disease has been investigated and compared with control. Short-term electrocardiograms (ECGs) were assessed in supine rest and active standing, and spectral measures of heart rate variability were determined. RESULTS: Two types of response to the change of supine to standing posture were determined in both healthy and diseased subjects. In majority of subjects, the increased heart rate induced by standing was accompanied by a decrease in high-frequency power. However, in about 30% of all subjects, increased heart rate during standing was accompanied by an increased high frequency power. Independently of posture and disease, high-frequency and low frequency power were positively correlated. In subjects characterized by a reduction of heart rate variability in standing, the high-frequency power in both postures is reduced in diseased subjects compared to control. CONCLUSIONS: These results demonstrate that in this age range, the response to posture is not unique because of the difference in high-frequency power, which implies a variety of vagal modulations of heart rate. PMID- 15882676 TI - Impact of noncoronary cusp prolapse in addition to right coronary cusp prolapse in patients with a perimembranous ventricular septal defect. AB - BACKGROUND: Few data are available with regard to the impact of aortic cusp herniation on the evolution of aortic regurgitation (AR) in patients with a perimembranous ventricular septal defect (VSD). METHODS: One hundred and two patients with a perimembranous ventricular septal defect with right coronary cusp prolapse were divided to two groups depending on the development of aortic regurgitation. The original defect diameter, the right coronary cusp deformity index (RCCD), and the right coronary cusp imbalance index were obtained as we reported previously. RESULTS: Mild aortic regurgitation was detected in 35 patients, and moderate in three. No aortic regurgitation was observed in 64 patients. A significantly larger number of patients had noncoronary cusp prolapse and the right coronary cusp imbalance index >/=1.30 in the aortic regurgitation group than in the no regurgitation group. Relative risk and odds ratio of noncoronary cusp prolapse and the right coronary cusp imbalance index >/=1.30 for development of aortic regurgitation were 3.69 (95% CI, 0.91-15.03) and 27.90 (95.94-130.85), and 2.23 (0.83-5.98) and 4.70 (1.44-15.27), respectively. Surgical closure was performed in 29 patients. All patients with no noncoronary cusp prolapse underwent simple patch closure of the ventricular septal defect, while five patients with noncoronary cusp prolapse and aortic regurgitation underwent aortic valvuloplasty. Among these, one patient needed aortic valve replacement. CONCLUSIONS: The additional complication of noncoronary cusp prolapse is a strong risk factor for the development of aortic regurgitation in patients with a perimembranous ventricular septal defect with right coronary cusp prolapse. PMID- 15882677 TI - Clinical course and complications of infective endocarditis in patients growing up with congenital heart disease. AB - BACKGROUND: Although a high number of patients with congenital heart disease (CHD) undergo surgical palliation or definite correction up to adolescence, adult congenital heart disease (ACHD) may remain a potential lifelong risk factor for infective endocarditis (IE) in patients growing up with congenital heart disease (GUCH). METHODS: In a retrospective case study of a tertiary care center long term clinical course and complications of patients with IE and GUCH were analysed. RESULTS: Data of 52 patients with CHD, who fulfilled the Saiman criteria for infective endocarditis and were treated between April 1986 and March 2001, were identified: Risk factors for infective endocarditis were previous cardiovascular operation (51.9%), use of foreign material (38.5%), dental or other surgical procedures without recommended antibiotic prophylaxis (25.0%), or cardiac catheterization (5.8%). Staphylococcal (38.9%) or streptococcal species (35.2%) were cultivated in most cases as causative microorganisms. Complications were: recurrence of IE (7.7%), septic embolisms (30.8%) leading to central nervous complications (7.7%), embolism of pulmonary arteries (7.7%), renal arteries (1.9%), arteries of the extremities (9.6%), or infarction of spleen (1.9%). Other cardiac (23.1%) or extracardiac (13.5%) complications were frequent. The need of re-operations during or after IE was high (67.3%). The hospital mortality was 1.9%, late mortality was 7.7%. CONCLUSIONS: Patients with IE and CHD show a broad clinical spectrum of cardiac and extracardiac complications. They may lead to a complicative short- and long-term course with the potential risk of death and a high number of re-operation. Efforts have to be made to improve long-term outcome of patients with ACHD by an interdisciplinary cooperation. PMID- 15882678 TI - Failure to achieve recommended LDL cholesterol levels by suboptimal statin therapy relates to elevated cardiac event rates. AB - OBJECTIVES: The majority of patients with myocardial infarction (MI) and hypercholesterolaemia does not achieve guideline recommended low-density lipoprotein cholesterol (LDL) levels. Suboptimal dosages of statins explain this dilemma in most patients. DESIGN AND SETTING: We evaluated the relationship between statin treatment quality (optimal: LDL<115 mg/dl, suboptimal: LDL>/=115 mg/dl, no statin therapy despite hypercholesterolaemia) and the subsequent incidence of coronary events (coronary death, nonfatal MI, bypass surgery) over a 30 months follow-up in a large cohort of post MI patients with hypercholesterolaemia (n=2045). Analysis was performed in a nested case-control manner comparing 173 cases with a coronary event and 346 matched controls. RESULTS: Patients who developed a coronary event were treated optimally in 11.0%, suboptimally in 43.4% (p<0.05 vs. optimal treatment) and were untreated in 45.7% (p<0.001 vs. optimal treatment). Respective numbers in event-free patients were 21.4%, 47.7%, and 30.9%. After adjustment for most potential confounders, including all cardiovascular risk factors and medication, the relative risk of future non-fatal MI and coronary death associated with a suboptimal statin treatment was 2.02 (95% CI 1.04 to 4.18) compared to optimal statin treatment. Moreover, the statin equivalent dose in optimally treated individuals was significantly higher than in suboptimally treated individuals (0.85+/-0.03 vs. 0.78+/-0.02, p<0.05). CONCLUSION: In this community-based study, a lipid lowering therapy with statins into the recommended target range of LDL levels may be associated with decreased cardiovascular risk compared to a statin therapy without titrating the LDL level below 115 mg/dl. Thus, the quality of statin treatment was identified as an independent predictor of coronary events in post MI patients. PMID- 15882679 TI - Pattern of exercise-induced ST change is related to coronary flow reserve in patients with chest pain and normal coronary angiogram. AB - OBJECTIVE: To evaluate the usefulness of exercise treadmill test in determining the true microvasculature-induced ischemia, we compared the pattern of ST depression with coronary flow reserve (CFR) using transthoracic Doppler echocardiography (TTE) in patients with chest pain and normal coronary angiogram. DESIGN: Fifty-nine subjects (M/F=21:38, mean age 55+/-9 years) with chest pain and normal coronary angiogram underwent maximal symptom-limited exercise treadmill test (ETT). CFR was estimated with TTE and dipyridamole. Patients with a history of acute myocardial infarction, regional wall motion abnormalities, hypertrophic cardiomyopathy, ejection fraction less than 50%, or primary valvular heart disease were excluded from this study. RESULTS: No ST change was observed in 20 of 59 (34%) patients, up slope depression was observed in 20 (34%), flat depression in 13 (22%), and down slope depression in 6 (10%). Eleven of thirty nine (28%) exercise positive patients had decreased CFR <2.1. CFR was 3.1+/-0.6 in group with no ST change, 3.1+/-0.6 in group with up slope depression, 2.1+/ 0.6 in group with flat depression (p<0.05 versus group with no change and group with upslope depression, respectively), and 2.0+/-0.4 in group with down slope depression (p<0.05 versus group with no change and group with up slope depression, respectively). Flat to down slope depression of ST change during ETT had sensitivity of 58% and specificity of 95% for predicting CFR <2.1. CONCLUSION: Flat and down slope depression of ST segment during ETT might increase the sensitivity and specificity to detect the true microvasculature induced ischemia that is defined as CFR less than 2.1 in patients with chest pain and normal coronary angiogram. PMID- 15882680 TI - Pseudo-cardioversion of atrial fibrillation. PMID- 15882681 TI - Celestial acronyms. PMID- 15882682 TI - Double right coronary artery with anomalous origin of septal arteries from the right coronary sinus. PMID- 15882683 TI - Heterotopic heart transplantation. PMID- 15882684 TI - An unusual complication of cardiac catheterization. PMID- 15882685 TI - Short-term fluctuations in sinus cycle length after premature ventricular beats in patients with hypertrophic cardiomyopathy and myocardial infarction. AB - BACKGROUND: Sinus cycle length has been reported to fluctuate after a ventricular premature beat (VPB). The purpose of this study was to assess the short-term fluctuations of sinus cycle length in patients with hypertrophic cardiomyopathy (HCM) and prior myocardial infarction (MI). METHODS: The relative deviation of RR intervals from the mean of the last two RR intervals preceding a VPB were calculated during the 20 subsequent beats following the VPB from Holter recordings in 92 patients with non-obstructive HCM, 57 patients with prior MI and 54 healthy controls. RESULTS: In controls, the deviations of the RR intervals were negative for several beats after a VPB and subsequently changed to positive before returning to the baseline. Similar changes in RR intervals following a VPB were exhibited in HCM patients; however, the late positive deviations of RR intervals were more marked than in controls. By contrast, in patients with prior MI, the early negative deviations of RR intervals were smaller compared with controls, and the deviations returned to the baseline without incidence of the positive changes. CONCLUSIONS: Short-term fluctuations in sinus cycle length after a VPB differed exclusively among HCM patients, prior MI patients, and healthy controls. PMID- 15882686 TI - Internal mammary artery steal syndrome secondary to an anomalous lateral branch. AB - A 53-year-old male who underwent three-vessel coronary artery bypass grafting had a left internal mammary artery (LIMA) graft to the left anterior descending artery (LAD) and saphenous venous grafts to right coronary artery (RCA) and left circumflex coronary artery. Four years after surgery, he developed exertion angina associated with upper body exercises and even deep breathing at times. Angiographic evaluation revealed an anomalous lateral internal thoracic artery with steal phenomenon documented by adenosine cardiolyte. Patient was successfully treated with transcutaneous steel coil embolization by closing the anomalous vessel. Repeat stress electrocardiogram did not show any signs of ischemia. This case report emphasizes the variability in internal mammary artery (IMA) anatomy and the need to completely ligate all the branches of internal mammary artery intraoperatively. PMID- 15882687 TI - This DAFNE is definitely not that DAFNE. PMID- 15882688 TI - Catecholamine therapy inducing dynamic left ventricular outflow tract obstruction. AB - Hypertrophic obstructive cardiomyopathy with significant hypertrophy of the basal septum is the most frequently reported cause of left ventricular outflow tract (LVOT) obstruction. Additionally, other conditions such as dehydration, sepsis, vasodilatation, or mitral valve repair have been associated with LVOT obstruction. In this report, we present a case of a patient without hypertrophy who developed severe dynamic left ventricular outflow tract obstruction during catecholamine stimulation for shock that complicated severe pancreatitis. The present case serves as a reminder that hypovolemia together with a hyperdynamic state resulting from catecholamine administration may result in the development of dynamic LVOT obstruction even if baseline cardiac evaluation is unremarkable. Early detection and intensive efforts to reverse the underlying conditions, including cessation of catecholamine therapy and correction of hypovolemia are essential. PMID- 15882689 TI - Magnetic resonance angiography in the assessment of a first reported case of duplicate right coronary artery. PMID- 15882690 TI - Catheter-induced straightening of external iliac tortuosity: a cause of pseudostenosis to be borne in mind. AB - Reversible vascular obstructive lesions, i.e. pseudostenoses, may pose significant threats to interventional cardiologists as they can be mistaken for obstructive lesions and prompt inappropriate revascularization procedures. We hereby report for the first time in the literature a case of external iliac artery pseudostenosis due to catheter straightening of significant underlying vessel tortuosities. Despite the initial angiographic image obtained from retrograde catheterization of the right external iliac artery which was strongly suggestive for significant stenosis, a thorough review of clinical history, physical examination and a second-look angiography by means of contralateral catheterization and contrast injection showed the absence of any significant lesion in the tortuous left external iliac artery, thus avoiding an unnecessary and potentially harmful vascular intervention. This clinical vignette emphasizes the importance of a thorough clinical examination and angiographic assessment for the appropriate diagnosis and management of reversible stenoses. PMID- 15882691 TI - Percutaneous transluminal angioplasty and stent implantation for coronary subclavian steal syndrome: unusual approach. PMID- 15882692 TI - Right ventricle hydatid cyst requiring tricuspid valve excision. AB - Hydatid cyst in the heart is uncommon, with cardiac involvement occurring in about 0.4-2% of all hydatid infestations. The right ventricle is only rarely involved accounting for 8% of cases. We report a case of a hydatid cyst located in the right ventricle involving the posterior papillary muscle requiring tricuspid valve excision. PMID- 15882693 TI - A life history model of somatic damage associated with resource acquisition: damage protection or prevention? AB - A resource acquisition-allocation model is developed to examine the trade-off between reproduction and somatic protection. Unlike previous studies, resource intake is not assumed to be constrained: instead, resource intake is free to vary, with increased intake being associated with an increased risk of somatic damage. This gives rise to an optimal resource intake as well as an optimal allocation strategy. This paper studies the relative importance of acquisition and allocation strategies in regulating acquisition-related mortality. Under the optimal allocation strategy mortality rate increases with age, in accordance with the disposable soma theory of aging. Contrary to the usual interpretation of the disposable soma theory, this increase in mortality can arise from an increase in the resource acquisition effort rather than a decrease in the resources allocated to protection. At early ages resource acquisition is found to be the primary path for regulating life history costs, whilst allocating resources to protection becomes more important later in life. Models for targeted and non-targeted damage repair are considered and the robustness of our results to the structure and parameterization of the model is discussed. The results from our models are discussed in light of published data. Resource acquisition is shown to be a potentially important mechanism for controlling somatic damage which deserves further study. PMID- 15882694 TI - On the optimality of the genetic code, with the consideration of coevolution theory by comparison of prominent cost measure matrices. AB - Statistical and biochemical studies have revealed non-random patterns in codon assignments. The canonical genetic code is known to be highly efficient in minimizing the effects of mistranslation errors and point mutations, since it is known that when an amino acid is converted to another due to error, the biochemical properties of the resulted amino acid are usually very similar to those of the original one. In this study, using altered forms of the fitness functions used in the prior studies, we have optimized the parameters involved in the calculation of the error minimizing property of the genetic code so that the genetic code outscores the random codes as much as possible. This work also compares two prominent matrices, the Mutation Matrix and Point Accepted Mutations 74-100 (PAM(74-100)). It has been resulted that the hypothetical properties of the coevolution theory of the genetic code are already considered in PAM(74-100), giving more evidence on the existence of bias towards the genetic code in this matrix. Furthermore, our results indicate that PAM(74-100) is biased towards the single base mistranslation occurrences in second codon position as well as the frequency of amino acids. Thus PAM(74-100) is not a suitable substitution matrix for the studies conducted on the evolution of the genetic code. PMID- 15882695 TI - Scaling properties of childhood infectious diseases epidemics before and after mass vaccination in Canada. AB - The goal of this paper is to analyse the scaling properties of childhood infectious disease time-series data. We present a scaling analysis of the distribution of epidemic sizes of measles, rubella, pertussis, and mumps outbreaks in Canada. This application provides a new approach in assessing infectious disease dynamics in a large vaccinated population. An inverse power law (IPL) distribution function has been fit to the time series of epidemic sizes, and the results assessed against an exponential benchmark model. We have found that the rubella epidemic size distribution and that of measles in highly vaccinated periods follow an IPL. The IPL suggests the presence of a scale invariant network for these diseases as a result of the heterogeneity of the individual contact rates. By contrast, it was found that pertussis and mumps were characterized by a uniform network of transmission of the exponential type, which suggests homogeneity in the contact rate or, more likely, boiled down heterogeneity by large intermixing in the population. We conclude that the topology of the network of infectious contacts depends on the disease type and its infection rate. It also appears that the socio-demographic structure of the population may play a part (e.g. pattern of contacts according to age) in the structuring of the topology of the network. The findings suggest that there is relevant information hidden in the variation of the common contagious disease time-series data, and that this information can have a bearing on the strategy of vaccination programs. PMID- 15882696 TI - On the role of endothelial progenitor cells in tumor neovascularization. AB - The exact role that bone marrow (BM)-derived endothelial progenitor cells (EPCs) play in tumor neovascularization is heavily debated. We develop a quantitative three-compartment model with predictive power regarding the dynamics of tumorigenesis. There are two distinct processes by which tumor neovasculature can be built: angiogenesis is the formation of new blood vessels from preexisting vessels; vasculogenesis is the formation of new vessels by recruiting circulating EPCs. We show that vasculogenesis-driven and angiogenesis-driven tumors grow in different ways. (i) If angiogenesis is the prevailing process, then the tumor mass (and volume) will grow as a cubic power of time, and BM-derived EPCs will stay at a constant level. (ii) If vasculogenesis is the dominant process, then the tumor mass will be characterized by a linear growth in time, and the number of circulating EPCs (after possibly increasing to a maximum) will decrease to low levels. With this information, one can identify the "signature" of each of the processes in the observations of tumor growth and the dynamics of the relevant characteristics, such as the level of BM-derived EPCs. We show how our results can help explain some apparently contradictory experimental data. We also propose ways to couple this study with directed experiments to identify the exact role of vasculogenesis in tumor progression. PMID- 15882697 TI - A computational method to detect epistatic effects contributing to a quantitative trait. AB - We develop a new computational method to detect epistatic effects that contribute to a complex quantitative trait. Rather than looking for epistatic effects that show statistical significance when considered in isolation, we search for a close approximation to the quantitative trait by a sum of epistatic effects. Our search algorithm consists of a sequence of random walks around the space of sums of epistatic effects. An important feature of our approach is that there is learning between random walks, i.e. the control mechanism that chooses steps in our random walks adapts to the experiences of earlier random walks. We test the effectiveness of our algorithms by applying them to synthetic datasets where the phenotype is a sum of epistatic effects plus normally distributed noise. Our test statistic is the rate of success that our methods achieve in identifying the underlying epistatic effects. We report on the effectiveness of our methods as we vary parameters that are intrinsic to the computation (length of random walks and degree of learning) as well as parameters that are extrinsic to the computation (number of markers, number of individuals, noise level, architecture of the epistatic effects). PMID- 15882698 TI - The histone octamer influences the wrapping direction of DNA on it: Brownian dynamics simulation of the nucleosome chirality. AB - In eukaryote nucleosome, DNA wraps around a histone octamer in a left-handed way. We study the process of chirality formation of nucleosome with Brownian dynamics simulation. We model the histone octamer with a quantitatively adjustable chirality: left-handed, right-handed or non-chiral, and simulate the dynamical wrapping process of a DNA molecule on it. We find that the chirality of a nucleosome formed is strongly dependent on that of the histone octamer, and different chiralities of the histone octamer induce its different rotation directions in the wrapping process of DNA. In addition, a very weak chirality of the histone octamer is quite enough for sustaining the correct chirality of the nucleosome formed. We also show that the chirality of a nucleosome may be broken at elevated temperature. PMID- 15882699 TI - Reproductive correlation and mean-variance scaling of reproductive output for a forest model. AB - We analyse the mean-variance scaling of reproductive output for a previously published forest model. The model relates individual reproductive effort and pollen limitation to the degree of synchrony in reproduction throughout a forest. We show that the exponent of Taylor's power law reflects the degree of synchrony of reproduction because it indicates the covariance of reproductive behavior. Further, we are able to relate the three components of masting, individual variability, population variability and synchrony in reproductive output, using Taylor's power law. Therefore Taylor's power law can be used as a synoptic index of masting. PMID- 15882700 TI - Statistical mechanics of myosin molecular motors in skeletal muscles. AB - Statistical mechanics provides the link between microscopic properties of matter and its bulk properties. The grand canonical ensemble formalism was applied to contracting rat skeletal muscles, the soleus (SOL, n = 30) and the extensor digitalis longus (EDL, n = 30). Huxley's equations were used to calculate force (pi) per single crossbridge (CB), probabilities of six steps of the CB cycle, and peak muscle efficiency (Eff(max)). SOL and EDL were shown to be in near equilibrium (CB cycle affinity 2.5 kJ/mol) and stationary state (linearity between CB cycle affinity and myosin ATPase rate). The molecular partition function (z) was higher in EDL (1.126+/-0.005) than in SOL (1.050+/-0.003). Both pi and Eff(max) were lower in EDL (8.3+/-0.1 pN and 38.1+/-0.2%, respectively) than in SOL (9.2+/-0.1 pN and 42.3+/-0.2%, respectively). The most populated step of the CB cycle was the last detached state (D3) (probability P(D3): 0.890+/ 0.004 in EDL and 0.953+/-0.002 in SOL). In each muscle group, both pi and Eff(max) linearly decreased with z and statistical entropy and increased with P(D3). We concluded that statistical mechanics and Huxley's formalism provided a powerful combination for establishing an analytical link between chemomechanical properties of CBs, molecular partition function and statistical entropy. PMID- 15882701 TI - Stochastic evolutionary dynamics on two levels. AB - We consider a population that is subdivided into groups. Individuals reproduce proportional to their fitness. When a group reaches a certain size it has a probability to split into two groups while another group is eliminated. In this stochastic process, the number of groups is constant, while the total population size fluctuates between well-defined bounds. We calculate the fixation probability of newly introduced mutants under constant selection. We show that the described population structure acts as a suppressor of selection compared to an unstructured population of the same size. The maximum suppression of selection is obtained, when the number of groups equals the number of individuals per group. We also study opposing selection on two or more levels by analysing the evolutionary dynamics of hierarchically embedded Moran processes. PMID- 15882702 TI - Autotoxicity in Lolium rigidum: analyzing the role of chemically mediated interactions in annual plant populations. AB - Autotoxicity is the chemical influence of one plant on the development of individuals of the same species. Autotoxicity in perennial plants can be a useful mechanism to avoid future conspecific competitors, however, the ecological role of autotoxicity in annuals, if any, remains uncertain. In this paper, we analyse the autotoxic phenomenon in Lolium rigidum, an annual, self-incompatible grass from semi-arid environments suspected to be autotoxic, and we dissert on the mechanisms of intraspecific interference, carrying out density-dependent experiments with additions of conspecific competitors and plant residues. Our experiments show that chemical interference and resource competition are density dependent processes that act in opposite directions. Inhibitory chemical interference increases at low population densities whereas competition for resources intensifies at high densities. Based on these results, the manuscript theorizes on the ecological role of the autotoxic phenomenon in short-lived plants. We hypothesize that autotoxicity enhances population viability by restraining the development of populations with few individuals, which should decrease the chances of outcross and experience inbreeding depression. If this hypothesis proves true, it could contribute to explain the unusually high within population diversity encountered in this species, which contrasts with reported values from other similar wind-pollinated, self-incompatible grasses. PMID- 15882703 TI - Evolution of mechanoregulation of bone growth will lead to non-optimal bone phenotypes. AB - Mechanical forces acting on the bones during growth affect their final shape and strength. Mechanoregulation of bone growth may be recognized in embryogenesis, and also in the adaptation of the adult skeleton to changes in mechanical loading. Mechanoregulatory responses for tissues have arisen during evolution, but does evolution give rise to responses that produce optimal skeletal phenotypes? In this paper, we investigate the emergence of an optimal mechanoregulation response in a population. By combining equations describing long bone growth with a genetic algorithm to describe evolutionary change, we created a computational model to simulate the evolution of mechanoregulation in bone growth. A population of individuals is created where each individual is assigned a diploid gene set which controls the growth and remodelling of the bone. At maturity, each bone is assessed and its 'fitness' calculated; fitness is quantified as bone strength relative to bone mass. The simulation continues for many generations, and includes mutations and a varying environment. The genes present in the population are tracked and the evolution of parameters governing mechanoregulation is calculated. The results indicate that a population may converge to one bone growth algorithm but, more usually, a range of mechanoregulation algorithms for different individuals will persist after many generations. Even if the population converges to one mechanoregulation law, convergence to the 'optimum' bone was never found. Although many researchers propose that natural selection has pushed skeletal structure towards an optimum, our computational model suggests that this is unlikely to be the case. PMID- 15882704 TI - Power-law scaling in dimension-to-biomass relationship of fish schools. AB - Motivated by the finding that there is some biological universality in the relationship between school geometry and school biomass of various pelagic fishes in various conditions, I here establish a scaling law for school dimensions: the school diameter increases as a power-law function of school biomass. The power law exponent is extracted through the data collapse, and is close to 35. This value of the exponent implies that the mean packing density decreases as the school biomass increases, and the packing structure displays a mass-fractal dimension of 53. By exploiting an analogy between school geometry and polymer chain statistics, I examine the behavioral algorithm governing the swollen conformation of large-sized schools of pelagics, and I explain the value of the exponent. PMID- 15882705 TI - Robustness and fragility of Boolean models for genetic regulatory networks. AB - Interactions between genes and gene products give rise to complex circuits that enable cells to process information and respond to external signals. Theoretical studies often describe these interactions using continuous, stochastic, or logical approaches. We propose a new modeling framework for gene regulatory networks, that combines the intuitive appeal of a qualitative description of gene states with a high flexibility in incorporating stochasticity in the duration of cellular processes. We apply our methods to the regulatory network of the segment polarity genes, thus gaining novel insights into the development of gene expression patterns. For example, we show that very short synthesis and decay times can perturb the wild-type pattern. On the other hand, separation of time scales between pre- and post-translational processes and a minimal prepattern ensure convergence to the wild-type expression pattern regardless of fluctuations. PMID- 15882706 TI - Laparoscopic donor nephrectomy. PMID- 15882707 TI - Robotic surgery and telesurgery in urology. PMID- 15882708 TI - Inguinal hernia in stage M0 prostate cancer: a comparison of incidence in men treated with and without radical retropubic prostatectomy--an analysis of 1105 patients. AB - OBJECTIVES: To analyze the incidence of inguinal hernia (IH) in a large group of patients with nonmetastatic prostate cancer who were treated nonoperatively, and to compare it with the incidence in a subset of patients who had undergone radical retropubic prostatectomy (RRP). IH has been reported in 12% to 21% of men at a mean period of 6 to 10 months after RRP. However, whether IH truly represents a complication after RRP has been somewhat debatable owing to the lack of proper control groups. METHODS: A total of 953 patients treated without surgery (nonoperative group) and 152 patients who underwent RRP (operative group) were selected from the Scandinavian Prostate Cancer Group Study No. 6 database consisting of 1218 patients with nonmetastatic prostate cancer. Radiotherapy, cryotherapy, and a follow-up duration of less than 3 months were exclusion criteria. Patients were followed up for any new medical condition at 12-week intervals for a mean period of 39 months (nonoperative group) and 50 months (operative group). RESULTS: Of the 953 patients in the nonoperative group, 23 (2.4%) developed IH versus 13 (8.6%) of 152 in the operative group (log-rank [Mantel-Cox] P = 0.010). CONCLUSIONS: Within comparable age groups, the incidence of IH in men with prostate cancer treated without surgery was significantly lower than that after RRP. This phenomenon seems to be causally related to the surgical procedure. The increased risk of IH after RRP deserves further recognition and should be included in the preoperative information given to patients. Studies are warranted to define the causal mechanisms, as well as possible preventive measures. PMID- 15882709 TI - Cost and time benefits of dual implantation of inflatable penile and artificial urinary sphincter prosthetics by single incision. AB - OBJECTIVES: To evaluate the efficiency, safety, and cost-effectiveness of synchronous prosthetic treatment of male urinary incontinence and impotence using a single transverse scrotal incision. METHODS: A total of 92 inflatable penile prostheses (IPPs), 21 artificial urinary sphincters (AUSs), and 15 combined IPPs/AUSs were implanted in 128 men at Brooke Army Medical Center and the University of Texas Health Science Center at San Antonio. The operative times and outcomes were compared among three groups (group 1, IPP; group 2, AUS; and group 3, dual IPP/AUS). We performed cost estimates of synchronous versus two-stage implant procedures. RESULTS: Dual implantation in a single-stage procedure significantly reduced (24.7%) the operative time (P <0.05, mean 113 minutes) compared with the total time for the individual procedures (IPP, average of 78 minutes; AUS, average of 72 minutes; total 150 minutes). No prosthetic infections or erosions occurred in this series. Dual implantation was associated with approximately a $7000 cost savings compared with individual procedures. CONCLUSIONS: The results of our study have shown that dual prosthetic implantation through a single incision is safe, efficient, and cost-effective. PMID- 15882710 TI - Incidence of upper tract abnormalities in patients with neurovesical dysfunction secondary to multiple sclerosis: analysis of risk factors at initial urologic evaluation. AB - OBJECTIVES: To determine the incidence of upper tract abnormalities on renal ultrasonography in patients with multiple sclerosis (MS) referred for urologic evaluation, as well as to identify any risk factors present on the basis of the historical information and urodynamic findings. METHODS: Data were derived from all patients with MS referred to the neurourology clinic during a 4-year period. The database was specifically queried for patients found to have upper tract abnormalities on screening renal ultrasonography. Demographic parameters, as well as laboratory values (creatinine) and urodynamic results, were evaluated for risk factors associated with abnormal upper tract findings. RESULTS: Of the 113 patients referred and evaluated, 66 completed both urodynamic testing and renal ultrasonography. Eleven (16.7%) had abnormal ultrasound findings, with focal caliectasis the most common finding. No demographic parameter (age, sex, time since MS diagnosis, MS pattern) was associated with a greater likelihood of abnormal renal ultrasonography on univariate analysis. Neither serum creatinine nor any urodynamic finding (including the presence of dyssynergia or the threshold and amplitude of detrusor overactivity) was associated with abnormal renal ultrasound findings. CONCLUSIONS: No patients in our series had any indication of obstructive uropathy more severe than mild hydronephrosis. Of the 16.7% of patients with any abnormal findings, most were noted to have minor caliectasis, likely to be of little clinical significance. Although no factors identifying patients at risk of renal abnormalities at presentation were found, ongoing evaluation of patients with baseline findings will serve to identify those at risk of progression. PMID- 15882711 TI - High kidney stone risk in men working in steel industry at hot temperatures. AB - OBJECTIVES: To study the incidence of urinary lithiasis and metabolic alterations among male employees from a steel industry who were exposed to high temperatures in the work environment. METHODS: A retrospective cross-sectional study was performed and consisted of two stages. First, the incidence of urolithiasis among the industry's 10,326 employees was assessed. These employees were divided into two group's: group 1 (n = 1289) consisted of the hot-area workers (temperature greater than 45 degrees C) and group 2 (n = 9037) consisted of those working in areas at room temperature. In the second stage, 59 workers without urolithiasis who underwent a metabolic evaluation were divided into two group's: group 3 (n = 34) consisted of hot-area workers and group 4 (n = 25) consisted of those working in areas at room temperature. Evaluations were made of calcium, creatinine, and uric acid in serum; in the 24-hour urine samples, we assessed the volume, calcium, uric acid, citrate, and oxalate. RESULTS: Of the 10,326 workers, 181 (1.75%) had presented with at least one episode of urinary stones. Of these, 103 were among the hot-area workers (8.0%) and 78 among the room-temperature workers (0.9%; P <0.001). The metabolic evaluation showed that the hot-area group (group 3), compared with the room-temperature group (group 4), presented more frequently with hypocitraturia (55.8% versus 28%, P = 0.03) and low urinary volume (79.4% versus 48%, P = 0.01). CONCLUSIONS: Workers exposed to high temperatures presented with a ninefold risk of lithiasis. Hypocitraturia and low urine volumes were the metabolic alterations observed. PMID- 15882712 TI - Laparoscopic live donor nephrectomy has equivalent early and late renal function outcomes compared with open donor nephrectomy. AB - OBJECTIVES: To assess the patterns of early functional recovery and long-term function in laparoscopic and open procured live donor nephrectomy (LDN) kidneys, highlighting the radionuclide scan as an additional tool for assessment, because of concerns regarding renal functional recovery after laparoscopic LDN. METHODS: We reviewed the donor and recipient records of 101 laparoscopic and 35 open LDNs performed between August 1997 and September 2001. Data were collected on demographic, immunologic, and intraoperative variables and ureteral/vascular complications. Delayed renal function recovery in recipients was evaluated by serum creatinine greater than 2.5 mg/dL on postoperative day 5, dialysis in first postoperative week, and two renographic criteria--the time to peak activity and the time to one-half peak activity on postoperative day 5. Long-term outcomes were evaluated by serum creatinine at 1, 3, 6, and 12 months and 2 and 3 years, creatinine clearance at 1 year, and patient and allograft survival. RESULTS: Donor and recipient age, sex, body mass index, and number of HLA mismatches did not differ between the two groups. The mean operating room time and blood loss were comparable. No differences were found in the early functional parameters (renography, creatinine at postoperative day 1 and 5, or dialysis in week 1) or long-term outcome (patient and graft survival, creatinine, and rejection at 1 year and patient and graft survival at 1, 2, and 3 years). CONCLUSIONS: Early recovery of graft function, longer term renal function, and 3-year patient and allograft survival are similar for live donor kidneys obtained by either a laparoscopic or an open surgical technique. PMID- 15882713 TI - Routine insertion of ureteral stent in live-donor renal transplantation: is it worthwhile? AB - OBJECTIVES: To evaluate the impact of the routine use of double-J stents in live donor renal transplantation at a single institute from a prospective randomized study. METHODS: A total of 100 patients were prospectively randomized into two groups of 50 patients each. Group 1 received a routine double-J silicone ureteral stent and group 2 did not. A standard Lich-Gregoir ureteroneocystostomy was performed in both groups. In group 1, the patients were scheduled for stent removal after 2 weeks. RESULTS: Both groups were comparable in terms of age, sex, ischemia time, number of renal arteries, and time to diuresis. In group 1, two grafts were lost in the early postoperative period and those patients were excluded from the final analysis. None of our patients in either group had developed a ureteral stricture at a mean follow-up of 10.8 +/- 3.6 months. In the stented group, 2 patients developed a urinary leak, but no leakage was reported in the nonstented group (P = 0.14). Although 19 patients in group 1 (39.6%) had a urinary tract infection, only 9 in group 2 (18%) showed evidence of a positive urine culture (P = 0.02). The presence of a ureteral stent and female sex were the independent predictors of postoperative urinary tract infection on multivariate analysis. The mean serum creatinine at discharge was 1.2 +/- 0.3 mg% and 1.2 +/- 0.4 mg% in groups 1 and 2, respectively (P = 0.2). CONCLUSIONS: The results of our study have shown that routine ureteral stent insertion has no impact on the rate of vesicoureteral leakage or obstruction in live-donor renal transplantation, whereas it is significantly associated with an increased incidence of urinary tract infection. Stenting should be limited to patients with a pathologic and/or defunctionalized bladder. PMID- 15882714 TI - Development of exophytic tumor model for laparoscopic partial nephrectomy: technique and initial experience. AB - OBJECTIVES: To develop and test a porcine model to help teach the techniques needed to perform laparoscopic partial nephrectomy (LPN), which is a technically challenging procedure with necessary reconstructive skills that are difficult to transfer. METHODS: A tumor model was created by unilateral subcapsular percutaneous injection of liquid plastic (Smooth-Cast 320) in five pig kidneys. Five Washington University urologists performed LPN and assessed the efficacy of the tumor model. Subsequently, the tumor model was evaluated as a tool for teaching LPN during the Washington University Advanced Laparoscopic and Robotic Urologic Oncology Course. Twenty-eight participants performed unilateral porcine LPN with the tumor model. Questionnaires were used to assess the utility of this tumor model. RESULTS: Unilateral tumors were successfully created in five pigs and remained intact during all LPN procedures. Visually, the tumors appeared as white exophytic masses. Ultrasonography revealed a well-circumscribed, hypoechoic lesion and a mean diameter of 2.02 cm. The mean operative time was 32.4 minutes. In subsequent testing, 24 (86%) of the 28 participants returned the questionnaire, and 96% responded that the tumor model had enhanced their LPN learning experience. Seven course participants (29%) reported problems with hemostasis, ultrasonography, or laparoscopic instrumentation. Two tumor model related complications occurred. During the initial evaluation, one pig experienced a fatal pulmonary embolism of the plastic. During the course, a second animal experienced extravasation of the solution into the renal collecting system. CONCLUSIONS: For surgical education purposes, the Smooth-Cast model is an effective surgical tool for LPN. Most of the surgeons in this evaluation believed the model enhanced their learning experience. PMID- 15882715 TI - Short-term efficacy of temperature-based radiofrequency ablation of small renal tumors. AB - OBJECTIVES: To present our experience using radiofrequency ablation (RFA) for the treatment of small renal tumors. Our objective was to assess the short-term (1 to 3 years) oncologic efficacy of RFA. METHODS: Consecutive renal tumors treated since May 2001 with a minimal follow-up of 6 months were included. Patients were treated with a temperature-based radiofrequency generator and were followed up with serial imaging at 6 weeks, 3 and 6 months, and every 6 months thereafter. RESULTS: A total of 109 small renal tumors (91 patients) were treated with computed tomography-guided percutaneous RFA (n = 63) or laparoscopic RFA (n = 46). The mean tumor size was 2.4 cm (range 0.8 to 4.7). The initial ablation was successful in 107 (98%) of 109 tumors. The two incomplete ablations were successfully re-ablated. Of the 60 patients with at least 1 year of follow-up, 60% had biopsy proven renal cell carcinoma (an additional 24% had no tissue diagnosis). In this group, one local recurrence (1.7%) was detected during a mean follow-up of 19.4 months (range 12 to 33), and in those with known renal cell carcinoma, none had evidence of distant progression (0%). The local recurrence was successfully re-ablated such that all 109 cases had no clinical or radiographic evidence of disease at last follow-up. Three patients died of causes unrelated to cancer. CONCLUSIONS: The results of our study have shown that in the short term, RFA appears to be a reasonable therapeutic nephron-sparing approach for treating select patients with small renal tumors. The cancer control appears adequate to date, but longer follow-up is necessary before widespread application. PMID- 15882716 TI - Laparoscopic management of peripelvic renal cysts: University of California, San Francisco, experience and review of literature. AB - OBJECTIVES: To report our experience and review published reports on the laparoscopic management of peripelvic renal cysts. Peripelvic renal cysts represent a unique subset of renal cysts, as they are rare, commonly symptomatic, and more difficult to treat than simple peripheral renal cysts. Minimally invasive methods for the treatment of peripelvic renal cysts, including laparoscopic decortication, have recently become more common. METHODS: Four patients who presented with symptomatic peripelvic cysts underwent laparoscopic decortication at our institution. All four were men aged 47 to 65 years. One patient had undergone an unsuccessful prior cyst aspiration. All patients underwent preoperative computed tomography and retrograde pyelography. The mean number of peripelvic cysts per patient was 3.0, and the mean cyst size was 7.1 cm. RESULTS: The mean operative time was 259 minutes (range 240 to 293), and the mean estimated blood loss was 30 mL (range 10 to 50). No evidence of cystic renal cell carcinoma was found on aspiration cytology or cyst wall pathologic examination. The mean hospital stay was 1.3 days. No inadvertent collecting system injuries and no intraoperative or postoperative complications occurred. All 4 patients achieved symptomatic relief and were determined to have radiologic success as determined by the 6-month postoperative computed tomography findings. CONCLUSIONS: Laparoscopic ablation of peripelvic renal cysts is more difficult than that of simple peripheral renal cysts and demands a heightened awareness of potential complications and, therefore, more advanced surgical skills. In addition to our experience, a thorough review of published reports found this procedure to be safe and effective with appropriate patient selection. PMID- 15882717 TI - Complex laparoscopic partial nephrectomy for renal hilar tumors. AB - OBJECTIVES: To evaluate our experience with laparoscopic partial nephrectomy (LPN) for tumors located adjacent to the renal hilum. Continued advances in laparoscopic technology and technique have made LPN feasible for increasingly anatomically complex tumors. METHODS: A retrospective chart review was performed of all patients who underwent LPN at Washington University. We identified 8 patients who had undergone LPN between December 2001 and September 2004 for hilar tumors that were defined as those located within 5 mm of the renal hilar vessels. The data were retrospectively analyzed for parameters, including operative time, morbidity, and postoperative course. RESULTS: LPN was successfully completed in all 8 patients without conversion to an open or hand-assisted approach. The indication for nephron-sparing surgery was elective in 6 patients and imperative in 2. The mean operative time was 3.0 hours (range 2.5 to 3.5), and the mean estimated blood loss was 188 mL (range 30 to 700). All patients had negative margins on the final pathologic examination. No intraoperative complications occurred. Nine postoperative complications developed in 6 patients. They included hemorrhage in 1, fever in 1, ileus in 1, urinary tract infection in 1, urine leak in 4, and transient postoperative neuropathy in 1 patient. CONCLUSIONS: With adequate laparoscopic experience, LPN for hilar tumors is a reasonable surgical option. In our experience, the procedure was associated with an increased risk of urine leak. Preoperative placement of a ureteral catheter to help delineate collecting system violations and routine postoperative stenting may reduce the incidence of this complication. PMID- 15882718 TI - Single-setting bilateral laparoscopic renal surgery: assessment of single-stage procedure. AB - OBJECTIVES: To assess the safety and efficacy of single-setting bilateral laparoscopic renal procedures. The continued advancement of laparoscopic surgical technology and surgeon expertise has allowed increasingly technically challenging procedures to be completed laparoscopically. Little has been reported on patient outcome, morbidity, and mortality of bilateral laparoscopic single-stage procedures. METHODS: Between May 2000 and February 2004, 13 patients underwent synchronous bilateral renal surgery. Both retroperitoneal (n = 5) and transperitoneal (n = 8) approaches were used. The data were retrospectively analyzed for operative time, morbidity, mortality, and postoperative course. RESULTS: Bilateral laparoscopic procedures were successfully completed in 11 (85%) of 13 patients. One patient required conversion to an open procedure because of significant adhesions. Another patient with von Hippel-Lindau disease had unexpected extensive pathologic features in each kidney and was therefore treated in a staged fashion. The mean operative time was 5.5 hours (range 4.7 to 8.5). The mean estimated blood loss was 268 mL (range 50 to 950). Patients resumed oral intake and ambulated within 24 hours after surgery. The mean analgesic requirement was 40.5 mg MSO4 equivalents (range 2 to 178). The mean hospital stay was 3.1 days (range 1 to 6). Patients returned to partial activity within the first week and enjoyed full activity at 3 weeks. One intraoperative complication and five postoperative complications occurred in 5 patients. CONCLUSIONS: Our results have demonstrated that single-setting bilateral laparoscopic renal surgery is safe and can expedite resolution of urologic pathologic findings without increased morbidity. Bilateral single-setting laparoscopic surgery should only be performed if the primary procedure has been completed expeditiously and without complications. PMID- 15882720 TI - Multicenter randomized clinical trial comparing surgery and collagen injections for treatment of female stress urinary incontinence. AB - OBJECTIVES: To compare, in a multicenter, randomized clinical trial, collagen injections versus surgery with regard to efficacy, quality of life, satisfaction, and complications. METHODS: Of 133 women with stress urinary incontinence, 66 were randomized to collagen injection and 67 to surgery (6 needle bladder neck suspensions, 19 Burch, and 29 slings). After randomization, 15 women refused their allocated treatment. "Intent-to-treat" and "per protocol" analyses were applied. Women assigned to collagen injection could receive up to three injections before it was considered a failure. A "top-up" injection was allowed within 3 months after cure. Success as the primary outcome at 12 months was defined as a dry 24-hour pad test (2.5 g or less of urine) after having received only the allocated intervention. RESULTS: The per protocol analysis showed that the success rate 12 months after collagen injections (53.1%) was much lower than that after surgery (72.2%). The difference was 19.1% (95% confidence interval 36.2% to -2%). The general and disease-specific quality-of-life scores measured by the Rand Medical Outcomes Study 36-item Health Survey and Incontinence Impact Questionnaire were similar in the two groups (P = 0.306). Women treated by surgery were, on average, more satisfied (79.6%) than those treated by collagen injection (67.2%), but the difference was not significant (P = 0.228). Finally, complications were less frequent and severe with collagen injection: 36 events in 23 subjects for collagen injection versus 84 events in 34 subjects for surgery (P = 0.03). CONCLUSIONS: One year after intervention, the success rate of collagen injection as a treatment for stress urinary incontinence was about 19% lower than that after surgery. This has to be tempered by the similar changes in quality of life and satisfaction in both groups and that the number and severity of complications were much greater after surgery than after collagen injection. The results of this study indicate that collagen injections might be a worthwhile alternative to surgery for the treatment of stress urinary incontinence. PMID- 15882721 TI - Early removal of nasogastric tube after cystectomy with urinary diversion: does postoperative ileus risk increase? AB - OBJECTIVES: To examine the risk factors of postoperative ileus and investigate whether the duration of postoperative nasogastric tube (NGT) use affects the rate of postoperative ileus in patients undergoing radical cystectomy with urinary diversion. METHODS: A total of 101 patients underwent radical cystectomy with urinary diversion from 1999 to 2003. We examined the demographic and perioperative variables of the patients who did and did not develop postoperative ileus. We divided the patients into two groups--those who had the NGT removed within 24 hours and those who had the NGT removed at first flatus. We compared the two groups for the incidence of ileus and clinical variables. RESULTS: Postoperative ileus was observed in 23 patients (23%). The demographic data of both groups were not different in terms of age or American Society of Anesthesiologists score. Also, the operative time, estimated blood loss, type of diversion, and postoperative complication rate were not significantly different between the two groups (P >0.05). However, the risk of postoperative ileus was significantly greater in those who took polyethylene glycol for bowel preparation than in those who took sodium phosphate (40% versus 18%, respectively, P = 0.02). No significant difference in the prevalence of ileus was found between the patients whose NGT was removed within 1 day and those in whom it was removed after 2 days (25% versus 22%, respectively, P >0.05). CONCLUSIONS: The results of our study suggest that the use of sodium phosphate for bowel preparation may reduce the incidence of postoperative ileus and that early NGT removal after cystectomy is not correlated with ileus. PMID- 15882722 TI - Cystoprostatectomy and orthotopic ileal neobladder reconstruction for management of bacille Calmette Guerin-induced bladder contractures. AB - OBJECTIVES: To evaluate the incidence of bacille Calmette-Guerin (BCG)-contracted bladders and attempt to ascertain the potential risk factors for this severe complication. BCG is the most effective intravesical agent for the treatment of high-grade, noninvasive transitional cell carcinoma (TCC) of the bladder. Widely used by urologists for the management of superficial TCC, adverse effects are not uncommon. These adverse effects are most commonly limited to irritative voiding symptoms, hematuria, and fever; however, severe adverse effects may occur. METHODS: We retrospectively reviewed two cystectomy databases (University of Southern California/Norris Cancer Center and University of Miami Miller School of Medicine) and identified patients who underwent cystectomy and orthotopic neobladder reconstruction for BCG-induced bladder contracture. RESULTS: We identified 1 case each in the two institutions' cystectomy databases of severely contracted bladders. The overall incidence of this complication leading to cystectomy was 0.05% and 0.3% at each institution. Both patients were salvaged with cystoprostatectomy and orthotopic reconstruction with an ileal neobladder. Neither patient had residual cancer at pathologic analysis of the cystectomy specimen, and both patients' severe lower urinary tract symptoms were alleviated postoperatively. CONCLUSIONS: A contracted bladder due to extravasation of BCG, although rare, is a serious complication. Cystoprostatectomy with orthotopic neobladder reconstruction may be the optimal solution to alleviate severe lower urinary tract symptoms and remove the risk of subsequent urothelial malignancy. PMID- 15882723 TI - Correlation between histologic findings and cytogenetic abnormalities in bladder carcinoma: a FISH study. AB - OBJECTIVES: To investigate the chromosomal abnormalities present in bladder carcinoma using a fluorescence in situ hybridization assay and to correlate the genetic findings with the pathologic grade and stage. METHODS: Samples from 37 bladder carcinomas were obtained at cystectomy or transurethral resection. In all cases, a histologic evaluation and fluorescence in situ hybridization analysis were performed. Pericentromeric DNA probes for chromosomes 7, 8, 9, and 17, and locus-specific DNA-probes for the 9p21 and 9q34 bands, were used in the fluorescence in situ hybridization assay. RESULTS: Grade 1-2 tumors were characterized by the loss of genetic material on chromosome 9 in 35.3% of cases and either no detectable alterations or multiple aneusomy events in 47.1% and 17.6% of the tumors, respectively. Polysomy was the most frequent occurrence in grade 3 and pT1-T4 carcinomas. No significant difference was found between the loss of 9p21, 9q34, or chromosome 9 and the different tumor classifications. A statistically significant difference was found in the frequency of polysomy between grade 1-2 and grade 3 tumors and between pTa and pT1-T4 tumors for chromosomes 7, 8, 9, and 17, as well as chromosome bands 9p21 and 9q34 (P <0.005). CONCLUSIONS: These findings show that chromosome 9 losses do not correlate with the tumor grade or stage, but are the only aberrations found at a significant frequency in low-grade lesions. The results suggest that pT1 tumors are closely related to muscle-invasive tumors at the genetic level and show that polysomy of the chromosomes assessed correlates with high-grade and high-stage bladder carcinoma. PMID- 15882724 TI - Differences between urologists in United States and Canada in approach to bladder cancer. AB - OBJECTIVES: To determine the Canada-United States differences with respect to the detection, diagnosis, surveillance, and treatment of bladder cancer. METHODS: A multiple-choice questionnaire was developed and mailed to 760 American and 516 Canadian urologists between November and December 2002. The areas assessed by the questionnaire included demographics, screening, superficial disease and recurrence, surveillance, muscle-invasive disease, advanced disease, and adjuvant systemic chemotherapy. RESULTS: The survey was adequately completed by 32.3% of American urologists and 40.0% of Canadian urologists (overall response rate 36.2%). Canadian urologists tended to be older and had larger practices than U.S. urologists (P <0.05). With respect to bladder cancer detection, U.S. urologists were more likely to use intravenous urography and cystoscopy than were Canadian urologists (P <0.0001). For patients with superficial disease, a significant proportion of urologists in both countries did not routinely use adjuvant chemotherapy. For surveillance, Canadian urologists performed cystoscopy (P <0.0001) and upper tract imaging (P <0.0001) less frequently than U.S. urologists. Striking differences were noted in the approach to Stage T2a disease, with U.S. urologists advocating radical cystectomy more frequently (P <0.0001). With respect to the type of urinary diversion, Canadian urologists tended to favor conduits (P <0.0001, male and P = 0.002, female). Canadian urologists were also less likely to use adjuvant chemotherapy among patients with advanced disease. CONCLUSIONS: The results of our study have shown that the trend of urologists in the United States is toward more aggressive screening, closer surveillance, an earlier trigger for cystectomy, and more common indications for intravenous chemotherapy. PMID- 15882725 TI - Additional use of [-2] precursor prostate-specific antigen and "benign" PSA at diagnosis in screen-detected prostate cancer. AB - OBJECTIVES: To evaluate the adjuvant clinical use of [-2] precursor prostate specific antigen ([-2]pPSA), which is associated with prostate cancer (PCa), and "benign" PSA, related to benign prostatic hyperplasia, in selecting a treatment strategy in patients with screen-detected PCa. METHODS: Research-use immunoassays (Beckman Coulter) were used to measure [-2]pPSA, sum [-7, -5, -4, and -2]pPSA, and benign PSA from the frozen serum of participants from the screen arm of the European Randomized Study of Screening for Prostate Cancer, section Rotterdam, diagnosed with PCa with a serum PSA level lower than 15 ng/mL. We compared men with relatively benign PCa (Epstein's criteria; group 1) and men with arbitrarily defined aggressive PCa characteristics (Gleason score greater than 4 + 4 and more than four cores with PCa invasion or pT3C disease; group 2). RESULTS: The data of 61 patients were evaluated. The median age in both groups was 68 years. Total PSA performed best in a univariate analysis, although in the multivariate analysis, the combination of pPSA and percent free PSA could correctly predict 95.5% of group 1 and 82.4% of group 2. The pPSA and percent free PSA forms remained statistically significant in the multivariate analysis of a subgroup of 30 participants normalized for PSA level and prostate volume; combined they correctly identified 89.5% and 54.5% of patients identified as having relatively favorable and aggressive PCa characteristics, respectively. CONCLUSIONS: Adjuvant clinical use of pPSA over traditional parameters in selecting treatment strategies for men with PCa cannot yet be definitely determined. However, the promising results in a subgroup analysis warrant further investigation. PMID- 15882726 TI - Prostate-specific antigen screening in a high-risk population: lessons from the community and how they relate to large-scale population-based studies. AB - OBJECTIVES: To estimate the incidence of prostate cancer among African-American men and Caribbean immigrants to the United States, to assess the applicability of large-scale prostate screening trials to a community screening program, and to recruit unscreened men. METHODS: African-American and Caribbean-American men were targeted with a community-based prostate cancer screening program in Jamaica, New York. Serum prostate-specific antigen determination and digital rectal examination were used to determine abnormal findings. The incidence of an abnormal screening examination was used to project the incidence of prostate cancer, which was compared with that in other reported trials. RESULTS: The projected incidence of prostate cancer among African-Americans and Caribbean Americans older than 50 years was 8% and 7%, respectively, similar to that reported in other trials of African-American men. The projected incidence of prostate cancer in Caribbean-American men aged 40 to 49 years was 1%, the same as the high rate reported among Caribbean men. As in other trials, a family history of prostate cancer and age were strong predictors of abnormal findings. Of the recruited men older than 50 years, 58% had never been screened compared with 42% nationally. CONCLUSIONS: Large population-based screening trials have identified ethnic groups at high risk of prostate cancer. This trial detected high rates of abnormal screening findings by targeting ethnicity. The incidence of an abnormal examination was high in Caribbean-American men younger than 50 years old. Finally, this trial successfully recruited underscreened men. PMID- 15882727 TI - Model to predict prostate biopsy outcome in large screening population with independent validation in referral setting. AB - OBJECTIVES: To develop a model capable of predicting prostate biopsy outcomes in a large screening population, with independent validation in the referral setting. METHODS: Data from 3814 men participating in the Tyrol screening project were used to develop the model. Prospectively collected data from two independent sites in the United States (Virginia Mason Clinic, Seattle, Wash and Stanford University, Stanford, Calif) were used to validate the model independently. The Tyrol data was split randomly into three cross-validation sets, and a feed forward, back error-propagation artificial neural network (ANN) was alternately trained on a combination of two of these data sets and validated on the remaining data set. Similarly, three logistic regression (LR) models were produced and validated using identical cross-validation data sets. The Tyrol model with the median area under receiver operating characteristic curve (AUROC) was then validated against the Virginia Mason (n = 491) and Stanford University (n = 483) data sets. RESULTS: The AUROCs for the three cross-validations were 0.74, 0.76, and 0.75 for the ANN and 0.75, 0.76, and 0.75 for the LR models. The mean AUROC for both ANN and LR was 0.75 with a standard deviation of 0.009 for ANN and 0.006 for LR. The AUROCs for the Virginia Mason and Stanford University data were 0.74 (both ANN and LR) and 0.73 (ANN) and 0.72 (LR), respectively. CONCLUSIONS: This model, designed to predict the prostate biopsy outcome, performed accurately and consistently when validated with data from two independent referral centers in the United States, suggesting that it generalizes well and may be of clinical utility to a broad range of patients. PMID- 15882728 TI - Prostate-specific antigen velocity accurately predicts response to salvage radiotherapy in men with biochemical relapse after radical prostatectomy. AB - OBJECTIVES: To determine whether prostate-specific antigen (PSA) velocity (PSAV), used as a selection criterion for salvage radiotherapy (RT) after radical prostatectomy (RP), predicts the likelihood of response to RT in men with biochemical relapse. METHODS: We retrospectively reviewed the records of 48 patients who had undergone salvage RT for biochemical relapse after RP. All men were followed up with serial PSA measurements for a minimum of 6 months from their initial PSA recurrence, and RT was only offered to those patients with a serum PSA level remaining at less than 1.0 ng/mL. The response to RT was defined as maintenance of a PSA level of less than 0.1 ng/mL. The pathologic and clinical parameters, including PSAV, were examined to determine their individual ability to predict the response to RT. RESULTS: Of the 48 patients, 30 had maintained a PSA level of less than 0.1 ng/mL at a median follow-up of 16 months. The PSAV was strongly predictive of the likelihood of a response to salvage RT. The median relapse-free survival time for patients with a PSAV of less than 0.035 ng/mL/mo was 28 months compared with 16 months for patients with a PSAV greater than 0.035 ng/mL/mo. All other parameters tested, including Gleason score, seminal vesicle invasion, extracapsular extension, and margin status, were not predictive of the likelihood of a response to RT. CONCLUSIONS: In the present study, PSAV accurately predicted the likelihood of response to salvage RT in men with biochemical relapse after RP. No other pathologic parameters predicted the likelihood of response to RT. Using PSAV as a sole selection criterion for salvage RT after RP may allow improvement in the historically low rates of durable response. PMID- 15882729 TI - Androgen deprivation for minimal metastatic disease: threshold for achieving undetectable prostate-specific antigen. AB - OBJECTIVES: To develop objective measures to select systemic therapies for study in large-scale trials for patients with lesser tumor burdens, we explored prostate-specific antigen (PSA) changes after androgen ablation in patients with disease progression after treatment of localized disease. Long-term follow-up of trials incorporating androgen-deprivation with local therapy have shown improved survival relative to local therapy alone. This suggests that the benchmark for treatment of minimal metastatic disease can be cure. METHODS: Patients with a rising PSA level with or without clinical metastases after local therapy who received androgen deprivation at Memorial Sloan-Kettering Cancer were identified from two institutional databases. The primary outcome was the proportion achieving an undetectable PSA level, and the pretreatment parameters associated with this endpoint were evaluated. RESULTS: A total of 130 patients who received androgen ablation and were followed up at Memorial Sloan-Kettering Cancer Center were identified. Overall, 31 (57%) of 54 (95% confidence interval 44% to 71%) patients with a rising PSA level alone and 28 (37%) of 76 (95% confidence interval 26% to 47%) patients with a rising PSA level and clinical metastases achieved an undetectable PSA level after androgen ablation (P = 0.02). The PSA level at the start of androgen ablation and the presence of metastases were the most significant predictive factors. CONCLUSIONS: The probability of achieving an undetectable PSA level varied inversely with the disease extent. Although achieving an undetectable PSA level does not mean that a patient has been cured, it does establish an endpoint that can be used to identify approaches worthy of study in the Phase III setting. PMID- 15882730 TI - Physical activity and sexual functioning after radiotherapy for prostate cancer: beneficial effects for patients undergoing external beam radiotherapy. AB - OBJECTIVES: To evaluate, in a cross-sectional study, the relationships among physical activity, sexual functioning, and treatment type for 111 men who had undergone radiotherapy for localized prostate cancer within the past 18 months. Physical activity preserves the sexual functioning capacity of older men. However, little information exists regarding the association of physical activity with sexual functioning after treatment for localized prostate cancer. METHODS: We tested the main effects of physical activity and treatment procedure, as well as their interaction, using hierarchical regression analysis. We hypothesized that greater physical activity would relate to better reported sexual functioning and that this relationship would be moderated by the type of medical treatment. RESULTS: After controlling for age, medical comorbidity, fatigue, and urinary and bowel functioning, more physical activity was significantly associated with better sexual functioning, and the interaction of physical activity and treatment procedure added a significant amount of explained variance. Overall, 35% of the variance in sexual functioning was accounted for by the model. Post hoc tests of moderation revealed that men who underwent external beam radiotherapy had significantly greater sexual functioning scores as physical activity increased but the effect of physical activity on sexual function after brachytherapy and combination therapy was nonsignificant. CONCLUSIONS: Physical activity was positively correlated with sexual functioning for those who underwent external beam radiotherapy. These relationships should be replicated and explored in a larger, longitudinal sample to ascertain whether the effects of physical activity in this at-risk population extend over time and protect men from treatment related decrements in sexual functioning. PMID- 15882731 TI - Laparoscopic extraperitoneal radical prostatectomy--learning curve of a laparoscopy-naive urologist in a community hospital. AB - OBJECTIVES: To describe the training approach that a laparoscopy-naive general urologist working in a nonteaching hospital used to successfully learn to perform laparoscopic extraperitoneal radical prostatectomy and describe the results obtained in the first 114 cases performed. METHODS: The urologist assisted an experienced laparoscopic surgeon for 20 extraperitoneal radical prostatectomies. During this time, he modified his technique of performing open radical retropubic prostatectomy to facilitate the acquisition of the laparoscopic techniques. Intracorporeal suturing was learned with the aid of a pelvic trainer. The clinical records of the first consecutive 114 cases were examined to evaluate the outcomes in terms of morbidity and oncological and functional concerns. A similar analysis was performed on a subgroup of 15 patients who had undergone laparoscopic extraperitoneal radical prostatectomy after previous transurethral resection of the prostate. RESULTS: The operating time progressively decreased during the learning curve. The mean duration of surgery was 160 minutes. Two conversions to open surgery were required owing to failure to progress. Of the 114 patients, 14% experienced complications, most of which were minor. The positive surgical margin rate was 17%. The average follow-up was 16 months. Undetectable serum prostate-specific antigen levels were observed in 82%, 87%, and 79% at 6, 12, and 18 months, respectively; 96% of patients used 0 to 1 pad per day for incontinence at 6 months of follow-up. The results in the transurethral resection subgroup were similar. CONCLUSIONS: Laparoscopic extraperitoneal radical prostatectomy can be successfully learned by a general urologist with no prior laparoscopic experience. PMID- 15882732 TI - Combined radical prostatectomy and bladder augmentation for concomitant prostate cancer and detrusor instability. AB - OBJECTIVES: To determine the outcomes of a select cohort of patients with severe voiding dysfunction, refractory to medical management, and a concomitant diagnosis of prostate cancer, who were treated with radical prostatectomy and augmentation enterocystoplasty. METHODS: Four men with biopsy-proven prostatic adenocarcinoma, as well as a diagnosis of severe overactive bladder, underwent combined radical retropubic prostatectomy and augmentation enterocystoplasty. All patients underwent fluorourodynamic testing confirming nonobstructive detrusor instability or hyperreflexia. Three patients underwent nerve-sparing radical retropubic prostatectomy with a clamshell ileocystoplasty, and one with neurogenic hyperreflexia underwent sigmoid cystoplasty with a continent catheterizable stoma at radical retropubic prostatectomy. RESULTS: The mean follow-up was 21.5 months (range 8 to 48). All patients had an undetectable prostate-specific antigen level postoperatively. The average hospitalization was 8 days. Perioperative complications occurred in 2 patients, including a prolonged urine leak managed with catheter drainage and postoperative hematuria requiring cystoscopic clot evacuation. Erectile function was preserved in 2 patients with good preoperative erections. At last follow-up, the 3 patients who voided per urethra had minimal postvoid residual urine volumes and maintained good continence, with only 1 patient describing occasional mild stress incontinence. At last follow-up, the patient with the sigmoid cystoplasty catheterized every 4 hours with volumes of about 300 mL and complete stomal continence. No patient required anticholinergic medications postoperatively. CONCLUSIONS: The concomitant diagnosis of prostate cancer and severe detrusor instability may be difficult to treat. The results of our study have shown that for those desiring surgical management for their prostate cancer, a combined bladder augmentation and radical prostatectomy may be performed with minimal added morbidity and significantly improved voiding function in the properly selected individual. PMID- 15882733 TI - Secondary azoospermia after vasovasostomy. AB - OBJECTIVES: To determine the frequency of secondary azoospermia after microsurgical vasovasostomy and to determine what factors increase the risk of its occurrence. METHODS: We performed a retrospective review of three surgeons' experience. Patency was defined as the presence of sperm in at least one postoperative semen sample. Transient patency was defined as azoospermia or no motile sperm after previous documentation of motile sperm. RESULTS: A total of 242 patients underwent 245 procedures (233 bilateral, 12 unilateral). The mean patient age was 39.2 +/- 0.4 years (range 24 to 56), and the mean obstructive interval was 8.7 +/- 0.3 years (range 0.25 to 24). The patency rates were 91% (224 of 245, sperm present in the semen) and 86% (208 of 245, motile sperm present in the semen). The transient patency rates were 5.3% (13 of 245, presence of sperm, with subsequent azoospermia) and 2.9% (7 of 241, motile sperm in the semen, with subsequent azoospermia) at a follow-up of 9.0 +/- 0.7 months (range 1 to 60). Transient patency occurred at a mean of 9.7 +/- 2.3 months. A greater risk of transient patency was observed with unilateral cases than bilateral cases (3 of 12, 25% versus 10 of 233, 4.3%; P = 0.0196, Fisher's exact test), and the obstructive interval was shorter for patent than for transient patent cases (Mann Whitney U test, P = 0.0458). CONCLUSIONS: The results of our study demonstrated that secondary azoospermia after vasovasostomy is rare. It is more common in unilateral cases and the obstructive interval for transiently patent cases is longer. Sperm cryopreservation, when motile sperm appear in the semen postoperatively, can circumvent the problem of secondary azoospermia, but most men will not need the frozen sperm. PMID- 15882734 TI - Extramammary Paget's disease of penis and scrotum. AB - OBJECTIVES: To make clear the uncertainty of the clinical outcome of extramammary Paget's disease (EMPD). Penile and scrotal involvement of EMPD is exceedingly rare, and only small series or case reports have been reported. METHODS: From 1995 to 2003, 36 patients with penile and scrotal EMPD were treated and followed up. Local wide excision was done in all patients with or without intraoperative frozen biopsy analysis. RESULTS: Of the 36 patients, 13 (36.1%) underwent intraoperative frozen biopsy analysis and only 1 patient (7.7%) had a positive surgical margin. However, 23 (63.9%) underwent local wide excision with excessive surgical margins of up to 1 to 2 cm only by gross examination, but 17 (73.9%) of them had positive surgical margins (P <0.01). Of the 17 patients with positive surgical margins, 8 developed local recurrence at a median of 8 months of follow up (P <0.05). One patient who had invasion to the subcutaneous tissue died of metastatic EMPD and internal malignancy (renal cell carcinoma) at 17 months after the initial operation. No patient had underlying adnexal carcinoma. CONCLUSIONS: The results of our study indicate that local wide excision with intraoperative frozen biopsy analysis is essential to the complete treatment of EMPD. PMID- 15882735 TI - Evaluation and management of genital anomalies in two patients with Klinefelter syndrome and review of literature. AB - OBJECTIVES: To present the evaluation and management of cryptorchidism and varicocele in patients with Klinefelter syndrome and review the literature. Recent advances in reproductive medicine have resulted in fertility in patients with Klinefelter syndrome. However, cryptorchidism and varicocele can affect the fertility potential on their own. METHODS: We present 2 cases and review the literature with an emphasis on the fertility potential. RESULTS: Germ cells were present in the testicular biopsy of an infant with cryptorchidism but not in the adolescent with a varicocele. The latter biopsy showed tubular sclerosis. CONCLUSIONS: The fertility potential of patients with Klinefelter syndrome and cryptorchidism may be preserved or enhanced by an early orchiopexy. However, the potential benefit of ligation of internal spermatic veins in patients with Klinefelter syndrome and a varicocele warrants further study. Cryopreservation should be considered early in adolescence or adulthood as germ cells become depleted at an accelerated rate after puberty. PMID- 15882738 TI - Genital trauma in children: classification and management. AB - OBJECTIVES: To determine the severity and, accordingly, the treatment of genital trauma in a pediatric population. METHODS: A total of 116 children with genital trauma and anorectal injury were retrospectively reviewed. The severity of trauma was graded according to the genital injury score (GIS), which we developed as a genital trauma scoring system. RESULTS: The median age was 8 years. Of the 116 children, 80 were girls and 36 were boys. The etiology of the trauma was traffic road accident (53 patients), fall, sexual abuse, and gunshot wound. Sixty-one patients had additional organ injuries. The GIS was I for 25 children, II for 19, III for 32, IV for 23, and V for 17. In addition to the primary repair, colostomy was performed in 22 patients. The most frequent postoperative complication was wound infection. The postoperative complication rate was significantly greater in patients with an injury severity score greater than 15, severe contamination, prolonged delay (longer than 8 hours), and a GIS of IV or V. CONCLUSIONS: The clarification of the mechanism and severity of the genital injury and associated organ injuries under general anesthesia may help in the appropriate classification. Primary repair should be the standard approach in genital trauma patients with a GIS of IV or less. Those with a GIS of V associated with severe contamination and prolonged delay require colostomy for improved outcome. PMID- 15882736 TI - Postnatal unilateral pelviureteral junction obstruction: impact of pyeloplasty and conservative management on renal function. AB - OBJECTIVES: To study the effect of pyeloplasty and conservative management on renal function in children with pelviureteral junction obstruction. METHODS: This prospective study included 65 children postnatally diagnosed with unilateral pelviureteral junction obstruction. On the basis of the patients' symptoms and technetium-99m diethylenetriamine pentaacetic acid renal isotope scan findings, symptomatic patients or those with split function of the corresponding kidney of 40% or less were assigned to group 1 (n = 35) and underwent pyeloplasty. Asymptomatic patients with split function greater than 40% (group 2, n = 30) were treated conservatively. Patients who showed deterioration of renal function underwent pyeloplasty. The effect of both lines of treatment on renal function was evaluated. RESULTS: After pyeloplasty, group 1 had improvement in the glomerular filtration rate (GFR) in 30 (85.7%), stabilization in 2 (5.7%), and deterioration in 3 (8.6%) patients. In group 1, the comparison between the mean value of the baseline GFR (17.1 +/- 7.7 mL/min) and its mean value at follow-up (28 +/- 6.3 mL/min) showed an increase of statistical significance (P <0.01). In the group 2 patients, conservative treatment resulted in improvement in the GFR in 12 (40%), stabilization in 3 (10%), and deterioration in 15 (50%) patients. In group 2, no significant difference was found between the mean baseline GFR (29.8 +/- 9.8 mL/min) and its mean value at follow-up (31.2 +/- 5.6 mL/min). CONCLUSIONS: In patients with postnatal pelviureteral junction obstruction, early pyeloplasty is recommended if the corresponding GFR is 40% or less. Conservative management in patients with better function resulted in deterioration of renal function in 50%. Should this policy of treatment be chosen, meticulous follow-up is mandatory. PMID- 15882739 TI - Novel endourologic applications for holmium laser. AB - The holmium laser is used routinely during endoscopic procedures to treat stones, strictures, and tumors of the urinary tract. We report two successful novel endourologic applications of a holmium laser: division and removal of a synthetic urethral sling mesh in one patient and fragmentation of a tempered stainless steel surgical needle in another. PMID- 15882740 TI - Feasibility study for robotic radical prostatectomy cautery-free neurovascular bundle preservation. AB - INTRODUCTION: Efforts continue to develop techniques that maintain the neurovascular bundles and minimize trauma for robotic laparoscopic radical prostatectomy. We evaluated the feasibility of preserving the nerve bundles without cautery or surgical clips. TECHNICAL CONSIDERATIONS: The seminal vesicles were dissected using scissors and bipolar cautery. After the rectum was mobilized, the vascular pedicles (VPs) were delineated. Laparoscopic bulldog clamps (30 mm) were placed at least 1 cm from the prostate. Using scissors, the VPs were divided right at the prostate. The neurovascular bundle was gently dissected off the prostatic capsule. After mobilizing the bundle, FloSeal was applied along its entire length. The FloSeal was then covered with a dry 1 x 4-cm sheet of Gelfoam. Once the prostate was removed, the bulldog clamps were sequentially withdrawn. The VPs were observed, and, if pulsatile bleeding was encountered, a 3-0 figure-of-eight suture was precisely placed for hemostasis. When hemostasis was complete, the anastomosis was performed. RESULTS: In 17 men, temporary vascular occlusion was applied to 27 VPs and FloSeal and Gelfoam was applied each time. In 4 cases (15%), hemostasis was inadequate because of continued arterial bleeding that was easily controlled with a superficial figure of-eight ligature of 3.0 absorbable suture. The average estimated blood loss was 91 mL (range 75 to 150). CONCLUSIONS: Cautery-free, clip-free, nerve-sparing robotic laparoscopic radical prostatectomy is feasible using a combination of temporary occlusion of the thick posterior prostatic pedicles with bulldog clamps followed by application of FloSeal. The effect on potency needs further follow up. PMID- 15882741 TI - Bladder exstrophy and phenotypic gender determination on fetal magnetic resonance imaging. PMID- 15882742 TI - Urethral hemangioma. PMID- 15882743 TI - Laparoscopic nephroureterectomy complicated by rupture of adrenal artery pseudoaneurysm. AB - We present a case of ruptured adrenal artery pseudoaneurysm as a complication of laparoscopic nephroureterectomy. An 87-year-old woman with hematuria and transitional cell carcinoma on imaging underwent laparoscopic nephroureterectomy. Her postoperative recovery was uneventful until day 17, when she presented with severe abdominal pain, syncope, and significant anemia. An aneurysm was suspected on computed tomography and Doppler ultrasonography. Selective angiography showed a pseudoaneurysm of the adrenal artery that could not be successfully embolized. Exploratory laparotomy obtained hemostasis, and the pseudoaneurysm was identified as involving an adrenal artery next to a surgical clip, which is speculated to be the cause. PMID- 15882744 TI - Metastatic renal cell carcinoma to contralateral ureter presenting as acute obstructive renal failure after radical nephrectomy. AB - Metastatic renal cell carcinoma commonly involves the lungs, bone, liver, adrenal glands, and brain. Less commonly affected organs include the gallbladder, thyroid, and pancreas. Even metastatic spread to the contralateral kidney and the bladder has been reported. Computed tomography is the standard imaging technique to evaluate for contralateral involvement. One of the disadvantages of computed tomography as a screening modality is its difficulty in identifying small ureteral lesions. We report a rare case of metastatic renal cell carcinoma in the contralateral ureter presenting as acute obstructive renal failure after radical nephrectomy. PMID- 15882745 TI - Unilateral testicular mass in man with chronic myelomonocytic leukemia: unusual presentation of chronic myelomonocytic leukemia sequela. AB - Extramedullary myeloid cell tumors are discrete tissue infiltrations by leukemic cells of myeloid lineage. They are more commonly associated with relapsing acute myelogenous leukemia but can occur in myeloproliferative/myelodysplastic leukemia, usually associated with disease acceleration. Although they can occur in any organ, reported testicular infiltration is rare. We describe the clinical presentation of an extramedullary myeloid cell tumor as a unilateral testicular mass in a man with known chronic myelomonocytic leukemia and its histologic diagnosis. To our knowledge, this manner of presentation is unique in this clinical context. PMID- 15882746 TI - Role of transrectal ultrasonography in diagnosis and treatment of retained Foley catheter. AB - Retention of Foley balloon in the bladder or urethra is a troublesome complication that presents a challenge to the urologist. We describe a complicated case of Foley catheter balloon retention (and trauma) within the urethra in an orthopedic patient undergoing hemiarthroplasty. The Foley balloon failed to deflate with standard methods. We used transrectal ultrasonography to successfully diagnose and manage the case. PMID- 15882747 TI - Intravascular extension of an osteosarcoma of the pubic bone into periprostatic venous plexus. AB - Pelvic osteosarcomas are uncommon but aggressive tumors that can invade the vascular system. We report a case of a 33-year-old man who presented with groin pain and was found to have a chondroblastic osteosarcoma invading the periprostatic venous plexus. Meticulous intraoperative exploration of the pelvis is necessary in such cases to resect tumor completely, thereby maximizing local control and preventing embolic complications. PMID- 15882748 TI - Rhabdomyosarcoma of tunica vaginalis masquerading as hydrocele. AB - Paratesticular rhabdomyosarcomas are rare tumors with aggressive growth patterns. Multimodal therapy with surgery, chemotherapy, and radiotherapy provides the patient with an excellent long-term prognosis. These tumors often present in the first two decades after birth. We report on the case of an 18-year-old man with a paratesticular rhabdomyosarcoma. PMID- 15882749 TI - Primary renal carcinoid tumor with liver metastases detected with somatostatin receptor imaging. AB - If the final pathologic examination reveals a carcinoid tumor of the kidney, additional evaluation to rule out another occult primary tumor site is necessary. If the primary origin of the tumor is confirmed to be from the kidney, additional evaluation is required for the detection of metastasis. Renal carcinoid tumors are extremely rare; however, both primary and metastatic renal carcinoid tumors have been reported. We report a case of a 40-year-old woman with primary carcinoid of the kidney and metastatic disease in the liver not evident by computed tomography and magnetic resonance imaging but identified by somatostatin receptor scintigraphy. PMID- 15882750 TI - Intravesical calculus secondary to ethanol gel injection into the prostate. AB - A 78-year-old man with coagulopathy and lower urinary tract symptoms secondary to benign prostatic hyperplasia underwent transurethral prostate ethanol gel injection. Six months after the injection, the patient presented with irritative symptoms and hematuria. Ultrasonography revealed a 5-cm intravesical calculus. Cystolitholapaxy was unsuccessful, and the patient underwent cystolithotomy with retrieval of a large, soft, round, brownish mass. Histologic examination of the mass demonstrated prostatic hyperplastic glandular stroma with extensive coagulate necrosis and peripheral calcification. To our knowledge, this is the first report of a calculus formed by prostatic tissue slosh after ethanol injection. PMID- 15882751 TI - Intracavernosal etilefrine self-injection therapy for recurrent priapism: one decade of follow-up. AB - Recurrent idiopathic priapism is a rare condition that, if not properly treated, may lead to impaired quality of life and erectile dysfunction. Treatment can be achieved by prevention of priapism episodes with systemic therapy or by early intervention with intracavernosal self-injection of sympathomimetic agents. We describe a case of a young patient with recurrent idiopathic priapism who has used etilefrine self-injection for the past 10 years with good efficacy and libido and erectile function preservation. This report suggests that this approach may be safely indicated in selected cases, particularly when sexual function preservation is a major concern. PMID- 15882752 TI - Identification of ectopic ureter in incontinent girl using magnetic resonance imaging. AB - We present the case of a 3.5-year-old girl who presented with continuous urinary leakage and a classic history suggesting an ectopic ureter, but an inconclusive physical examination and normal imaging by ultrasonography. Magnetic resonance imaging was finally able to identify an ectopic ureter draining a poorly functioning upper pole of a duplex system as the culprit. We discuss the increasing value of magnetic resonance imaging in identifying subtle pediatric genitourinary anomalies, such as in our case, and discuss the management options. PMID- 15882753 TI - Nonimmunosuppressant immunophilin ligand GPI-1046 does not promote in vitro growth of prostate cancer cells. AB - OBJECTIVES: Nonimmunosuppressant immunophilin ligands such as GPI-1046 have gained interest recently for their clinical potential to reduce the extent of injury sustained by penile innervation during radical prostatectomy for prostate cancer treatment and thereby improve erectile function recovery postoperatively. Preclinical studies in rat animal models of cavernous nerve injury have demonstrated that immunophilin ligands exert both neuroprotective and neuroregenerative effects on physiologic erection and on the morphology of erectile tissue and penile innervation. Before establishing their clinical roles, however, it would be useful to evaluate whether they exert mitogenic effects on malignant prostate cells. METHODS: Human prostate cancer cell lines (LAPC-4, CWR22Rv1, LNCaP, and PC-3) were treated in vitro with 0 to 10 microM of the prototypical nonimmunosuppressant ligand GPI-1046 in 1% fetal bovine serum (FBS) containing (ie, low growth) media, and their growth was assayed during a 7-day period using spectrophotometric cell counting. The results were normalized for comparison to the maximal growth produced in 10% FBS-containing media. RESULTS: All four prostate cancer cell lines grew maximally in 10% FBS-containing media, and this growth was reduced by more than twofold (P <0.05) when the cells were maintained in 1% FBS-containing media. The addition of 10 microM or less GPI-1046 to the cells in 1% FBS-containing media did not exert statistically significant mitogenic effects on human prostate cancer cells in vitro. CONCLUSIONS: These data contribute toward allaying concerns that the use of nonimmunosuppressant immunophilin ligand therapy for the improvement of erectile function recovery after radical prostatectomy will promote recurrent prostate cancer. PMID- 15882754 TI - Ethnic differences in neuroendocrine cell expression in normal human prostatic tissue. AB - OBJECTIVES: To determine whether any differences exist in neuroendocrine (NE) cell differentiation in normal prostates among various ethnic groups because the incidence and mortality from prostate cancer vary across racial groups. METHODS: Archived paraffin-embedded prostate samples not containing any malignancy were obtained from cystoprostatectomy specimens. Prostatic tissue was obtained from 15 African Americans, 13 Hispanics, 15 Asians, and 16 whites. NE cells were identified based on immunoreactivity for chromogranin A. The mean number of NE cells per high power field (HPF) for each patient was determined using a visual quantitative method by two observers who were unaware of the race of the patients. RESULTS: The geometric mean number of NE cells was 6.1/HPF for Asians, 5.6/HPF for whites, 4.0/HPF for Hispanics, and 0.7/HPF for African Americans. A highly significant difference was observed in the distribution of NE cells between African Americans and each of the other races (P < or =0.003). A trend toward greater NE expression was noted in Asians and whites compared with Hispanics; however, this difference did not reach statistical significance. CONCLUSIONS: We found a fivefold to eightfold difference in the distribution of NE cells in the normal prostates of African-American men compared with that of other races. The comparatively low NE cell expression in African-American men may play a role in the greater rate of prostate carcinogenesis seen in this ethnic group. PMID- 15882755 TI - Expression of CD44 and E-cadherin cell adhesion molecules in hypertrophied bladders during chronic partial urethral obstruction and after release of partial obstruction in rats. AB - OBJECTIVES: To determine the functional changes in the bladder and the expression of adhesion molecules in bladder tissue during chronic partial urethral obstruction and after release of partial obstruction in rats. METHODS: Twenty-one male Sprague-Dawley rats were separated into three groups, each containing 7 rats. A sham operation was performed in group 1 and cystometry was done 6 weeks later. In groups 2 and 3, hypertrophied unstable bladders were developed by partial infravesical outflow obstruction during a 6-week period. After this period, cystometry was performed in all group 2 rats. In group 3, the ligature was removed, the rats were followed up for 6 weeks, and then cystometry was performed. After cystometric evaluation, the bladders in all the rats were removed, weighed, and studied immunohistopathologically. RESULTS: After release of infravesical outflow obstruction, the bladder weight, residual volume, bladder capacity, maximal voiding pressure, voiding amplitude, and bladder contraction time decreased and bladder compliance increased in group 3 compared with group 2. CD44 and E-cadherin expression in the interstitial space and uroepithelial bladder tissue in group 2 rats stained intensely compared with those of groups 1 and 3. CONCLUSIONS: After release of 6 weeks of infravesical outflow obstruction, the cystometric parameters were significantly improved. Expression of CD44 and E cadherin in the obstructed bladder tissue may be a pathologic sign of inflammation. PMID- 15882756 TI - Effect of terazosin on tissue vascularity and apoptosis in transitional cell carcinoma of bladder. AB - OBJECTIVES: To present a pilot study to determine whether the alpha1-adrenoceptor antagonist terazosin can induce apoptosis in transitional cell carcinoma (TCC) of the bladder, similar to the effect seen with prostate cancer. The alpha1 adrenoceptor antagonist terazosin has recently been shown to induce apoptosis in prostate cancer cells both in vitro and in vivo and to reduce prostatic tissue vascularity by potentially affecting endothelial cell adhesion. METHODS: The records of 24 men who underwent radical cystectomy for TCC of the bladder at the Lexington Veterans Affairs Medical Center were reviewed. The control group consisted of 15 men who were never exposed to terazosin. The study group consisted of 9 men who were treated with terazosin before cystectomy. Sections of the bladder tumor and normal trigone were subjected to immunohistochemical analysis for microvessel density, endothelial cell CD31 expression, and apoptosis detection (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling), as well as high-molecular-weight cytokeratin staining. RESULTS: A significant reduction in tissue vascularity (14.0 versus 19.2, P <0.05) and a significant increase in the apoptotic index (3.0% versus 1.7%, P <0.05) was detected in terazosin-treated bladder tumors compared with untreated bladder tumors. Most TCC specimens (80%) exhibited strong and consistently uniform immunostaining for high-molecular-weight cytokeratin staining. CONCLUSIONS: These results suggest that terazosin reduces tumor vascularity and induces apoptosis in TCC of the bladder. Additional studies with more patients are necessary to reach definitive conclusions. However, considering the proven apoptotic action of terazosin in prostatic tissue, this study may have implications for the use of terazosin in the treatment of bladder TCC. PMID- 15882757 TI - Measurement of the mechanical characteristics of benign prostatic tissue: a novel method for assessing benign prostatic disease. AB - OBJECTIVES: To investigate the relationship between the morphology and the mechanical properties of benign prostatic tissues measured in vitro. METHODS: Fresh tissue specimens were collected from 17 patients undergoing transurethral resection of the prostate for benign prostatic obstruction. Individual tissue specimens underwent immediate mechanical testing, by applying a dynamic compressive strain to the samples. The amplitude ratio (E*) and phase difference (tan delta), measures of tissue elastic and viscous components, were derived. Individual sections from the processed specimens underwent immunohistochemical staining and computerized image analysis to measure the morphologic characteristics of each transurethral resection of the prostate chipping. Correlations between the morphologic and mechanical measurements were assessed. RESULTS: A strong positive correlation was found between prostatic smooth muscle content and (E*) (R2 = 0.58, P = 0.009). CONCLUSIONS: The results of this study have demonstrated that strong correlations exist between prostatic tissue morphology and mechanical characteristics. We believe that the ability to quantify prostatic tissue mechanical characteristics in vivo may be of clinical benefit in the future assessment and treatment of benign prostatic disease. PMID- 15882758 TI - Expression of nm23-H1 gene product in sarcomatous cancer cells of renal cell carcinoma: correlation with tumor stage and expression of matrix metalloproteinase-2, matrix metalloproteinase-9, sialyl Lewis X, and c-erbB-2. AB - OBJECTIVES: To investigate the clinical significance of nm23-H1 gene product in sarcomatous cancer cells, because this is known as a tumor-metastasis suppressor. Renal cell carcinoma with sarcomatoid cancer cells is characterized by high malignant potential and a poor prognosis. METHODS: We investigated the expression of nm23-H1 gene product in the carcinomatous and sarcomatous component (CC and SC) of renal cell carcinoma using immunohistochemical techniques and the relationships between the expression and clinicopathologic features. We also examined the expression of matrix metalloproteinase (MMP)-2, MMP-9, sialyl Lewis X, and c-erbB-2 in the SC because these proteins are regulated by the nm23-H1 gene or its products. RESULTS: We examined 20 renal cell carcinoma specimens that contained an SC and CC. The CC of 12 of the 20 tumors stained positively for nm23 H1 gene product. In contrast, the SC of only 3 of the 20 stained positive. The reduced expression of nm23-H1 gene product in the SC correlated significantly with tumor invasion (P <0.01), but not with tumor size or metastasis. In contrast, the expression of the nm23-H1 gene product in the CC was not associated with these pathologic features. Expression of the nm23-H1 gene product correlated negatively with MMP-2 expression (r = -0.48, P = 0.03). Other factors did not show such significant correlations with nm23-H1 gene product expression. CONCLUSIONS: Our results suggest that low expression of nm23-H1 gene products may play important roles in tumor invasion, and that this process is mediated in part by overexpression of MMP-2. PMID- 15882759 TI - Glutathione S-transferase T1-1 activity upregulated in transitional cell carcinoma of urinary bladder. AB - OBJECTIVES: To perform a systematic functional investigation of different glutathione S-transferase (GST) classes, including GST class Theta (GSTT) member GSTT1-1, in transitional cell carcinoma (TCC) and the surrounding normal uroepithelium of the same individuals. Recently, it was suggested that GSTT1-1 might be an important risk modulator for TCC. METHODS: Tumor samples and surrounding normal uroepithelium were obtained from 24 patients with TCC of urinary bladder. The following substrates with differential specificities were used: 1-chloro-2,4-dinitrobenzene for overall GST activity; 7-chloro-4-nitrobenzo 2-oxa-1,3-diazole for GST Alpha; 1,2-dichloro-4-nitro-benzene for GST Mu; 4 vinylpyridine for GST Pi 1-1(GSTP1-1); and 1,2-epoxy-3-(p-nitrophenoxy)propane for GSTT1-1. RESULTS: GSTP1-1 and GSTT1-1 activities were demonstrated in all uroepithelial and TCC samples, and GST Mu activity was detectable in 11 of 24 patients. In the tumor specimens, significant upregulation of all expressed GST subtypes was observed. The mean GSTP1-1 and GSTT1-1 level in TCC was increased 2 fold and 3.6-fold, respectively, compared with the mean level in the normal uroepithelium (P <0.001). Tumor GSTT1-1 activities correlated statistically significantly with the tumor stage (P <0.05). CONCLUSIONS: In tumors and adjacent normal uroepithelium of patients with TCC, three major cytosolic GST classes, Mu, Pi, and Theta, were expressed. Although the GST isoenzyme pattern in TCC was similar to that of the corresponding normal uroepithelium, during cancer progression a clear tendency toward an increase in all the GST subtypes expressed was noted. For the first time, distinct GSTT1-1 activity levels were demonstrated in human uroepithelium, as well as its pronounced upregulation in TCC. PMID- 15882760 TI - Social anxiety in primary care: hidden in plain view? PMID- 15882761 TI - Improving primary care for military personnel and veterans with posttraumatic stress disorder--the road ahead. PMID- 15882762 TI - Social anxiety disorder in primary care. AB - OBJECTIVE: To examine the prevalence, comorbidity, disability and mental health treatment associated with social anxiety disorder (SAD) in primary care, and to determine whether patients with SAD avoid seeking help from their primary care providers. DESIGN: We analyzed data from a health survey conducted on a systematic sample of patients. Data were then cross-linked to the practice's automated database in order to compare primary health care utilization by patients with SAD to that of patients with other psychiatric disorders and well controls. SETTING: Urban primary care practice at a teaching hospital. PATIENTS: A systematic sample (n=207) of primary care patients. MEASUREMENTS AND MAIN RESULTS: Patients were interviewed by mental health professionals using the Composite International Diagnostic Interview. Lifetime prevalence of SAD was 5.7%. Substance use disorders were far more common among patients with SAD than patients with other psychiatric disorders (33.3% vs. 3.3%, P=.01). Social anxiety disorder patients were functionally impaired and made fewer primary care visits per year (mean 4.1) compared to patients with other psychiatric disorders (mean 6.9; P=.016) or well controls (mean 6.4; P=.031); 41.7% reported receiving mental health treatment in the past year. CONCLUSION: Patients with SAD made fewer primary care visits compared to patients with other psychiatric disorders and well controls. These results, together with the high prevalence of substance use in SAD, and the finding that less than one half received past year mental health treatment, suggest substantial unmet need for care and are especially important in view of available effective treatments for SAD. PMID- 15882763 TI - Prevalence of posttraumatic stress disorder in Veterans Affairs primary care clinics. AB - Although posttraumatic stress disorder (PTSD) is relatively common in community epidemiologic surveys (5-6% for men, 10-12% for women), and psychiatric patients with PTSD are known to have poor functioning and high levels of psychiatric comorbidity, there are no studies that address PTSD prevalence, functioning, and burden in primary care settings. This article reports on (1) the prevalence of PTSD using Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition diagnostic criteria in Veterans Affairs (VA) primary care settings, (2) associated sociodemographic characteristics and comorbidities, (3) functional status related to PTSD, (4) the extent to which PTSD was recognized by providers and (5) health services use patterns (including specialty mental health) of PTSD patients. Patients were randomly selected from those who had an outpatient visit in FY 1999 at one of four VA hospitals; 888 patients consented (74.1% of 1198 contacted); 746 patients (84.0% of consenting patients; 62.3% of contacted patients) were reached for telephone diagnostic interviews. Diagnostic interviews with the Clinician Administered PTSD Scale yielded estimates of current PTSD prevalence of 11.5%. At statistically significant levels, PTSD was positively associated with a variety of comorbid psychiatric disorders, war zone service, age <65 years, not working, less formal education and decreased functioning. Of patients diagnosed with PTSD by study procedures, 12-month medical record review indicated that providers identified only 46.5% and only 47.7% had used mental health specialty services. PTSD-positive [PTSD(+)] patients who used mental health care in the past 12 months were more apt to be identified as having PTSD than nonmental health service users (78.0% vs. 17.8%). Although PTSD(+) patients had more medical record diagnoses than PTSD-negative [PTSD(-)] patients (6.28 vs. 4.95), their use of primary care, urgent care and inpatient care was not different from PTSD(-) patients. PMID- 15882764 TI - Negative affect states and cardiovascular disorders: a review and the proposal of a unifying biopsychosocial concept. AB - AIM: The purpose of this review was to study the relationships between negative affect states and cardiovascular disorders. PROCEDURE: The phenomenology of the negative affect states of depression, helplessness, hopelessness, vital exhaustion and grief is described. Their correlations with morbidity and mortality are analyzed. The physiological correlates of the affect states are pointed out. Finally, the reaction pattern of conservation-withdrawal according to Schmale and Engel and its ontogenesis are outlined. This is a disengaging behavior pattern as opposed to the engaging fight-flight reaction pattern of Cannon. The giving up complex, with its affects of helplessness and hopelessness, is explained. CONCLUSIONS: The giving up complex in the context of the conservation-withdrawal pattern presents a biologically and developmentally sound conceptual basis for the understanding of the relationships of the negative affect states with cardiovascular disorders. This enables the integration of the concept of vital exhaustion, which has become the most promising operationalized instrument in psychosocial cardiovascular research. PMID- 15882765 TI - Stability of the diagnosis of first-episode psychosis made in an emergency setting. AB - OBJECTIVE: This study aimed at evaluating the stability of the first psychotic episode diagnosis in the emergency context. METHODS: Fifty-nine patients were selected during a 15-month period and were followed for an average of 19.35 +/- 6.12 months. The admission and discharge emergency diagnosis were compared with the longitudinal diagnosis, obtained by the application of Structured Clinical Interview for DSM-IV Axis I Disorders--clinical version at the end of the follow up. Severity rating scales (Brief Psychiatric Rating Scale, Young Mania Rating Scale and Hamilton Rating Scale for Depression) were applied in the emergency assessment. RESULTS: Agreement between admission emergency diagnosis and longitudinal diagnosis was unsatisfactory (k=0.25), whereas that between emergency discharge and longitudinal diagnosis was satisfactory (k=0.57). Brief psychotic disorder diagnosis presented higher sensitivity rates but low specificity, comprising several false positives. Bipolar disorder had the highest rates of specificity. CONCLUSION: Brief psychotic disorder may not be a useful concept in the emergency assessment. A short period of observation can improve emergency psychiatric diagnosis. PMID- 15882766 TI - Impact on delirium detection of using a sensitive instrument integrated into clinical practice. AB - Early symptoms of delirium often go unnoticed. The Nursing Delirium Screening Scale (Nu-DESC) is a recently developed short, accurate and sensitive 24-h screening instrument. The Nu-DESC is more sensitive than the instrument from which it was derived, the Confusion Rating Scale (CRS). This study examined the impact on delirium detection of using the Nu-DESC over the CRS in 134 consecutive oncology patients. Expected false-negative rate (FNR) reductions at different delirium prevalence rates when using the Nu-DESC compared to the CRS and the number needed to screen (NNS) by the Nu-DESC were calculated. Kaplan-Meier survival analyses were used to study Nu-DESC-CRS divergences in delirium status and length of delirium-free survival. Ninety-nine patients were negative for delirium according to both tests. Of the remaining 35 patients, 16 had identical Nu-DESC-CRS delirium status and delirium-free survival, whereas 19 were detected later by the CRS (mean, 4.8 days). Among the 19 patients, 6 were still CRS negative upon hospital discharge. Integrating a continuous and sensitive delirium assessment instrument into usual care can facilitate its recognition, since more cases of delirium are diagnosed and patients are detected earlier. PMID- 15882767 TI - History of depression and women's current health and functioning during midlife. AB - OBJECTIVE: To examine the association of past depression with current physical and mood symptoms and functioning in a community cohort of middle-aged African American, White, and Hispanic women without current depression and whether the associations varied by severity of prior depression. METHODS: The study was conducted as part of a longitudinal multisite investigation of middle-aged women's health, the Study of Women's Health Across the Nation (SWAN). Nine hundred twenty-two women, aged 42-52 years, participated in The Structured Clinical Interview for the Diagnosis of DSM-IV Axis I Disorders (SCID) at study entry at three SWAN sites; 780 did not have current depression and formed the analytic sample. chi(2), ANOVAs and Cochran-Armitage Trend tests were conducted as appropriate to evaluate bivariate relationships between history of major depression and covariates and outcomes. Multivariable logistic regression analyses included significant covariates in final analyses. RESULTS: Women (24.3%) had a history of major depression: 14.9% single episode, 9.4% recurrent and 12.6% had minor depression. In multivariable logistic regression analyses, compared to no history of depression, any past depression predicted high body pain [odds ratios (ORs), 1.8-2.3; 95% CIs, 1.05-4.02]. Recurrent depression predicted poor social functioning (OR, 2.1; 95% CI, 1.20-3.80) and current treatment for back pain (OR, 4.2; 95% CI, 1.78-9.82). Minor depression predicted mood symptoms (OR, 1.9; 95% CI, 1.16-3.20). CONCLUSIONS: Midlife women with past major or minor depression are at risk for physical symptoms, body pain, and poor social functioning even in the absence of current depression. Primary care providers may underestimate the health impact of prior depression without current depression. PMID- 15882768 TI - Predictors and outcomes of delirium. AB - OBJECTIVES: To determine factors associated with the occurrence of delirium among patients undergoing surgical repair of abdominal aortic aneurysm (AAA). METHODS: The sample included all consenting patients who underwent AAA repair during a 12 month period. Before surgery, daily while in hospital, and at 1 and 6 months after surgery, we assessed patients' mood, mental status and functional status. We compared delirious and nondelirious patients for severity of preoperative depressive symptoms, length of hospital stay and mortality. The effects of delirium on postoperative functional status were assessed in conjunction with postoperative depressive symptoms using regression models. RESULTS: The sample of 35 patients was primarily male and elderly; one-quarter had three or more medical conditions; and eight (23%) developed delirium after surgery. Postoperative delirium was significantly associated with preoperative depressive symptoms, alcohol use and cognitive impairment as well as with longer lengths of stay and poorer functional status at 1 and 6 months after surgery. CONCLUSION: Identification and treatment of patients with depressive symptoms, alcohol use and cognitive impairment prior to AAA surgery could reduce the incidence of postoperative delirium and the prolonged hospital stays and impaired functional status associated with it. Surgeons should consider using simple screening instruments before surgery to identify patients at risk and referring them for psychiatric evaluation and treatment. They should also consider including psychiatrists early in the care of high-risk patients to improve detection of and early intervention for delirium. PMID- 15882769 TI - Dhat (semen loss) syndrome: a functional somatic syndrome of the Indian subcontinent? AB - OBJECTIVE: To study the illness characteristics of Dhat syndrome, which has been widely regarded as a culture-bound syndrome. METHODS: The sample was selected from a general hospital and included 29 patients with Dhat syndrome and 32 medical controls. Somatization Screening Index (SSI), Illness Behaviour Questionnaire (SIBQ), Somatosensory Amplification Scale (SAS), Whitley Index (WI) and Chalder Fatigue Scale were administered to both groups. RESULTS: Patients with Dhat syndrome scored significantly higher on all the measures except the SAS. DISCUSSION: The results indicate that patients with Dhat syndrome have significantly different illness beliefs and behaviours compared to controls and have similarities with other functional somatic syndromes (FSSs). PMID- 15882770 TI - Rapid oral loading of extended release divalproex in patients with acute mania. AB - BACKGROUND: Oral loading with the delayed release formulation of divalproex sodium is widely used for the treatment of patients with acute mania and produces rapid attainment of therapeutic serum levels. Recently, an extended release formulation of divalproex sodium (divalproex ER) was approved for treatment of migraine headaches. This formulation may be a useful treatment option for patients with acute mania. METHOD: A retrospective review of medical records was conducted on 14 inpatients with acute mania whose treatment included initiation of divalproex ER at a dose of 30 mg kg(-1) day(-1) in a single dose. Doses, serum levels and side effects associated with this treatment were recorded from the medical records of these patients. RESULTS: The average dose of divalproex ER was 2034 mg day(-1) (range, 1500-3000 mg day(-1)). Two of the 14 patients (14%) had documented side effects, none of which were severe. The average level obtained on day 3 after initiation of divalproex ER treatment was 93.2 mug ml(-1) (range, 47 136 mug ml(-1)), and in all but three patients valproate levels at this time point were within the therapeutic range of 50-125 mug ml(-1). In no case was divalproex ER discontinued due to a perceived lack of efficacy. CONCLUSION: The results suggest that divalproex ER can be safely administered by oral loading in inpatients with acute mania and that using a standard loading protocol can result in therapeutic serum levels in most patients in 3 days or less. PMID- 15882771 TI - 12,800-mg ziprasidone overdose without significant ECG changes. PMID- 15882772 TI - Possible serotonin syndrome with citalopram following cross-titration of clozapine to ziprasidone. PMID- 15882773 TI - The effect of milnacipran (serotonin noradrenaline reuptake inhibitor) on memory in Korsakoff's syndrome after encephalitis. PMID- 15882774 TI - Molecular mechanisms of dendritic spine development and remodeling. AB - Dendritic spines are small protrusions that cover the surface of dendrites and bear the postsynaptic component of excitatory synapses. Having an enlarged head connected to the dendrite by a narrow neck, dendritic spines provide a postsynaptic biochemical compartment that separates the synaptic space from the dendritic shaft and allows each spine to function as a partially independent unit. Spines develop around the time of synaptogenesis and are dynamic structures that continue to undergo remodeling over time. Changes in spine morphology and density influence the properties of neural circuits. Our knowledge of the structure and function of dendritic spines has progressed significantly since their discovery over a century ago, but many uncertainties still remain. For example, several different models have been put forth outlining the sequence of events that lead to the genesis of a spine. Although spines are small and apparently simple organelles with a cytoskeleton mainly composed of actin filaments, regulation of their morphology and physiology appears to be quite sophisticated. A multitude of molecules have been implicated in dendritic spine development and remodeling, suggesting that intricate networks of interconnected signaling pathways converge to regulate actin dynamics in spines. This complexity is not surprising, given the likely importance of dendritic spines in higher brain functions. In this review, we discuss the molecules that are currently known to mediate the exquisite sensitivity of spines to perturbations in their environment and we outline how these molecules interface with each other to mediate cascades of signals flowing from the spine surface to the actin cytoskeleton. PMID- 15882775 TI - Oxidative stress in the brain: novel cellular targets that govern survival during neurodegenerative disease. AB - Despite our present knowledge of some of the cellular pathways that modulate central nervous system injury, complete therapeutic prevention or reversal of acute or chronic neuronal injury has not been achieved. The cellular mechanisms that precipitate these diseases are more involved than initially believed. As a result, identification of novel therapeutic targets for the treatment of cellular injury would be extremely beneficial to reduce or eliminate disability from nervous system disorders. Current studies have begun to focus on pathways of oxidative stress that involve a variety of cellular pathways. Here we discuss novel pathways that involve the generation of reactive oxygen species and oxidative stress, apoptotic injury that leads to nuclear degradation in both neuronal and vascular populations, and the early loss of cellular membrane asymmetry that mitigates inflammation and vascular occlusion. Current work has identified exciting pathways, such as the Wnt pathway and the serine-threonine kinase Akt, as central modulators that oversee cellular apoptosis and their downstream substrates that include Forkhead transcription factors, glycogen synthase kinase-3beta, mitochondrial dysfunction, Bad, and Bcl-x(L). Other closely integrated pathways control microglial activation, release of inflammatory cytokines, and caspase and calpain activation. New therapeutic avenues that are just open to exploration, such as with brain temperature regulation, nicotinamide adenine dinucleotide modulation, metabotropic glutamate system modulation, and erythropoietin targeted expression, may provide both attractive and viable alternatives to treat a variety of disorders that include stroke, Alzheimer's disease, and traumatic brain injury. PMID- 15882776 TI - The role of acetaldehyde in the neurobehavioral effects of ethanol: a comprehensive review of animal studies. AB - Acetaldehyde has long been suggested to be involved in a number of ethanol's pharmacological and behavioral effects, such as its reinforcing, aversive, sedative, amnesic and stimulant properties. However, the role of acetaldehyde in ethanol's effects has been an extremely controversial topic during the past two decades. Opinions ranged from those virtually denying any role for acetaldehyde in ethanol's effects to those who claimed that alcoholism is in fact "acetaldehydism". Considering the possible key role of acetaldehyde in alcohol addiction, it is critical to clarify the respective functions of acetaldehyde and ethanol molecules in the pharmacological and behavioral effects of alcohol consumption. In the present paper, we review the animal studies reporting evidence that acetaldehyde is involved in the pharmacological and behavioral effects of ethanol. A number of studies demonstrated that acetaldehyde administration induces a range of behavioral effects. Other pharmacological studies indicated that acetaldehyde might be critically involved in several effects of ethanol consumption, including its reinforcing consequences. However, conflicting evidence has also been published. Furthermore, it remains to be shown whether pharmacologically relevant concentrations of acetaldehyde are achieved in the brain after alcohol consumption in order to induce significant effects. Finally, we review current evidence about the central mechanisms of action of acetaldehyde. PMID- 15882777 TI - Role of retinoid signalling in the adult brain. AB - Vitamin A (all-trans-retinol) is the parent compound of a family of natural and synthetic compounds, the retinoids. Retinoids regulate gene transcription in numerous cells and tissues by binding to nuclear retinoid receptor proteins, which act as transcription factors. Much of the research conducted on retinoid signalling in the nervous system has focussed on developmental effects in the embryonic or early postnatal brain. Here, we review the increasing body of evidence indicating that retinoid signalling plays an important role in the function of the mature brain. Components of the metabolic pathway for retinoids have been identified in adult brain tissues, suggesting that all-trans-retinoic acid (ATRA) can be synthesized in discrete regions of the brain. The distribution of retinoid receptor proteins in the adult nervous system is different from that seen during development; and suggests that retinoid signalling is likely to have a physiological role in adult cortex, amygdala, hypothalamus, hippocampus, striatum and associated brain regions. A number of neuronal specific genes contain recognition sequences for the retinoid receptor proteins and can be directly regulated by retinoids. Disruption of retinoid signalling pathways in rodent models indicates their involvement in regulating synaptic plasticity and associated learning and memory behaviours. Retinoid signalling pathways have also been implicated in the pathophysiology of Alzheimer's disease, schizophrenia and depression. Overall, the data underscore the likely importance of adequate nutritional Vitamin A status for adult brain function and highlight retinoid signalling pathways as potential novel therapeutic targets for neurological diseases. PMID- 15882778 TI - The taste cell-related diffuse chemosensory system. AB - Elements expressing the molecular mechanisms of gustatory transduction have been described in several organs in the digestive and respiratory apparatuses. These taste cell-related elements are isolated cells, which are not grouped in buds, and they have been interpreted as chemoreceptors. Their presence in epithelia of endodermal origin suggests the existence of a diffuse chemosensory system (DCS) sharing common signaling mechanisms with the "classic" taste organs. The elements of this taste cell-related DCS display a site-related morphologic polymorphism, and in the past they have been indicated with various names (e.g., brush, tuft, caveolated, fibrillo-vesicular or solitary chemosensory cells). It may be that the taste cell-related DCS is like an iceberg: the taste buds are probably only the most visible portion, with most of the iceberg more caudally located in the form of solitary chemosensory cells or chemosensory clusters. Comparative anatomical studies in lower vertebrates suggest that this 'submerged' portion may represent the most phylogenetically ancient component of the system, which is probably involved in defensive or digestive mechanisms. In the taste buds, the presence of several cell subtypes and of a wide range of molecular mechanisms permits precise food analysis. The larger, 'submerged' portion of the iceberg is composed of a polymorphic population of isolated elements or cell clusters in which the molecular cascade of cell signaling needs to be explored in detail. The little data we have strongly suggests a close relationship with taste cells. Morphological and biochemical considerations suggest that the DCS is a potential new drug target. Modulation of the respiratory and digestive apparatuses through substances, which act on the molecular receptors of this chemoreceptive system, could be a new frontier in drug discovery. PMID- 15882779 TI - Lack of evidence for association between the serotonin transporter gene (SLC6A4) polymorphisms and autism in the Chinese trios. AB - Serotonin regulates several aspects of brain development, and it is involved in a range of behaviors frequently disturbed in autistic disorder. The serotonin transporter is a critical component of the serotonergic system. The serotonin transporter gene (SLC6A4) is of special interest given the nature of the biological findings and the reported effects of selective serotonin reuptake inhibitors of autistic symptoms. So far the genetics researches of the SLC6A4 gene have given conflicting results. The aim of study was to investigate the association between the SLC6A4 gene and autism in the Chinese Han population. The present study was conducted with the detection of three single nucleotide polymorphisms (SNP(S)) located within the SLC6A4 gene by using the polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) analysis. We performed a family-based association study of these polymorphisms in 175 Chinese Han family trios. Linkage disequilibrium (LD) measurement (D') analysis showed the presence of LD between markers across the locus. No significant evidence of association was found at any of the markers detected by using the transmission disequilibrium test (TDT) and haplotype analyses in all samples and male samples. Our findings suggest that it is unlikely that DNA variations in the SLC6A4 gene play a significant role in the genetic predisposition to autism in the Chinese Han population or that allelic heterogeneity at the SLC6A4 loci dilutes potential disease-allele association. PMID- 15882780 TI - 5-HT2A-mu opioid receptor mechanisms in the hypothalamus mediate interleukin 1beta fever in rats. AB - Direct administration of interleukin-1beta (IL-1beta) into the lateral cerebral ventricle of rat brain, in addition to inducing febrile responses, upregulated the immunoreactivity of tryptophan hydroxylase in the preoptic anterior hypothalamus. The fever induced by IL-1beta was significantly attenuated by pretreatment with intracerebroventricular injection of 5-HT2A receptor antagonists including cyproheptadine, ketanserin, or mianserin. In addition, the IL-1beta-induced fever was mimicked by intracerebroventricular administration of a 5-HT2A receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-amionpropane (DOI). The DOI-induced (present results) or IL-1beta-induced (previous results) fever was further attenuated by pretreatment with an intracerebroventricular dose of mu opioid receptor antagonists (e.g., buprenorphine or cyclic d-phe-cys-Try-d-Arg Thr-pen-Thr-NH2) or 5-HT receptor antagonists (e.g., ketanserin or cyproheptadine). These findings suggest that a 5-HT2A-mu opioid receptor mechanism in the hypothalamus may mediate the IL-1beta fever. PMID- 15882781 TI - Morphine modulates glutamate release in the hippocampal CA1 area in mice. AB - Opiate abuse is associated with long-lasting neural adaptative changes in the brain. Increasing evidence demonstrates that opiates significantly alter the function of the glutamatergic system, while how the system is regulated still remains elusive. In the present study, we studied the effect of morphine on extracellular glutamate concentration in the hippocampus, a nucleus rich of the glutamatergic neurons. The results showed that glutamate concentration in the extracellular fluid of the hippocampus was decreased following either acute or chronic treatment of morphine. However, naloxone-induced withdrawal increased glutamate concentration significantly. These results suggest an adaptation of the glutamatergic neuronal transmission in the hippocampus after morphine treatment. PMID- 15882782 TI - Intrathecal application of cyproheptadine impairs locomotion in intact rats. AB - In intact adult rats, cyproheptadine, a 5-HT2 antagonist, administered intrathecally at the midlumbar segments was found to impair hindlimb locomotor movements during overground locomotion. These effects were dose-dependent; they varied from transient complete hindlimb paraplegia seen at doses of 300 microg/20 microl, to short-lasting trunk instability at doses of 100 microg/20 microl. After the return of overground locomotion, transient abduction of one of the hindlimbs was observed in some animals. These findings demonstrate that the blockade of 5-HT2 receptors affects locomotion in intact rats. Our results provide support for the hypothesis of serotonergic involvement in rat locomotion, which, so far, has been based mainly on the effects of 5-HT2 agonists on the recovery of locomotion in spinal rats. PMID- 15882783 TI - Ependymal epithelium disruption after vanadium pentoxide inhalation. A mice experimental model. AB - The blood-brain barrier (BBB) protects the CNS against chemical insults. Regulation of blood-brain tissue exchange is accomplished by ependymal cells, which possess intercellular tight junctions. Loss of BBB function is an etiologic component of many neurological disorders. Vanadium (V) is a metalloid widely distributed in the environment and exerts potent toxic effects on a wide variety of biological systems. The current study examines the effects of Vanadium pentoxide (V2O5) inhalation in mice ependymal epithelium, through the analysis of the brain metal concentrations and the morphological modifications in the ependymal cells identified by scanning and transmission electron microscopy after 8 weeks of inhalation, in order to obtain a possible explanation about the mechanisms that V uses to enter and alter the CNS. Our results showed that V2O5 concentrations increase from the first week of study, stabilizing its values during the rest of the experiment. The morphological effects included cilia loss, cell sloughing and ependymal cell layer detachment. This damage can allow toxicants to modify the permeability of the epithelium and promote access of inflammatory mediators to the underlying neuronal tissue causing injury and neuronal death. Thus, understanding the mechanisms of BBB disruption would allow planning strategies to protect the brain from toxicants such as metals, which have increased in the atmosphere during the last decades and constitute an important health problem. PMID- 15882784 TI - Neural correlates of eye blinking; improved by simultaneous fMRI and EOG measurement. AB - Neural correlates of eye blink in healthy human subjects can be investigated using functional magnetic resonance imaging. However, the focus of most previous studies has been on intentional eye blinking. The goal of the present study was to examine the neural correlates of spontaneous eye blinking with the help of EOG measurements during data acquisition of fMRI. After the removal of the pulse artifact in the EOG signal, EOG waveform clearly indicates eye blinking, which was equivalent to those measured outside of the MRI scanner. On the basis of this detection, each blinking event can be used as a temporal cue for the event related fMRI. In a comparison, we also investigated the neural correlates of blink inhibition. Based on the brain activation pattern, the activation of the bilateral parahippocampal, the visual cortex was commonly observed for both conditions. The additional activation of the precentral gyrus, corresponding to blink inhibition, and the right medial frontal gyrus, corresponding to spontaneous blinking were observed. Based on these results, we conclude that the medial frontal gyrus is responsible for spontaneous eye blinking, whereas precentral activation appears to be related to blink inhibition. PMID- 15882785 TI - Contribution of neck proprioception to subjective vertical perception among experts in physical activities and untrained women. AB - The purpose of this study was to investigate the influence of physical training on subjective vertical perception with the different head positions in order to explore the involving of the neck proprioception. Visual field dependence was assessed using a rod and frame test on women practising judo or dance (international level) or no specific physical activity. Tests were performed with head upright or tilted head to disturb the Z-axis egocentric reference frame. A cluster analysis determined the distribution of visual field independent (VFI) and visual field dependent (VFD) participants. The first result showed no head position effect for the group of judoists (6 degrees +/-5 degrees ; 7 degrees +/ 5 degrees ) and dancers (4 degrees +/-2 degrees ; 5 degrees +/-3 degrees ) but a significant effect for untrained participants (5 degrees +/-2 degrees ; 7 degrees +/-3 degrees ): their visual vertical perception was more disturbed with tilted head than with head upright. A variability exists among experts and would necessitate further analysis. The second result showed no influence of the head position for all VFD participants, whereas for VFI participants significant difference between upright and tilted head appeared both for experts (3 degrees +/-1 degrees ; 4 degrees +/-2 degrees ) and untrained participants (3 degrees +/ 1 degrees ; 5 degrees +/-2 degrees ). In this research, whatever physical activity level, the VFI participants would mainly use the Z-axis reference frame and rely on proprioceptive information. VFD among experts and VFI among untrained participants suggest that proprioceptive reference frame of neck may not provide alone according the groups an appropriate postural control. PMID- 15882786 TI - Lack of genetic association of cholesteryl ester transfer protein polymorphisms with late onset Alzheimers disease. AB - Dysregulation of cholesterol homeostasis may be associated with the pathogenesis of coronary artery disease (CAD) and Alzheimers disease (AD). Recently, several single nucleotide polymorphisms (SNPs) in cholesteryl ester transfer protein (CETP) were associated with altered plasma CETP concentrations, cholesterol concentrations and CAD. Hence, these CETP SNPs represent excellent candidates for evaluating association with AD. To date, one study has evaluated the association between a single CETP SNP and AD. In this study, we examined three CETP SNPs to evaluate the genetic association of CETP with late onset AD on two study cohorts: the Religious Orders Study (ROS) series, including 85 AD and 70 non-AD individuals, and the University of Kentucky (UKY) series, including 78 AD and 84 non-AD individuals. Significant association between CETP genotypes or haplotypes and late onset AD was not detected in these two study cohorts. Moreover, the CETP genotypes and haplotypes were not significantly associated with AD when the populations were stratified for the presence or absence of apolipoprotein E4 (apoE4). In summary, CETP genetic variants were not associated with AD in two series. PMID- 15882787 TI - Protein kinase C contributes to abnormal capsaicin responses in DRG neurons from cats with feline interstitial cystitis. AB - Interstitial cystitis (IC) is a painful disorder which affects urinary bladder function in cats and humans. We have used patch clamp techniques to examine the possibility that the properties of primary afferent neurons are changed in feline interstitial cystitis (FIC). We measured transient receptor potential vanilloid receptor 1 (TRPV1) responses to capsaicin (CAPS) in dorsal root ganglion (DRG) neurons (L4-S3) from normal cats and cats with FIC. We show that FIC neurons are increased in size and exhibit CAPS responses which are increased in amplitude and desensitize slowly. CAPS responses desensitized seven times slower in FIC neurons. Phorbol 12,13-dibutyrate (PDBu), an activator of PKC, slowed the desensitization of CAPS responses in normal cat bladder and non-bladder neurons, but had no effect in FIC neurons. Bisindolylmaleimide, an inhibitor of PKC, reversed the PDBu effects in normal cat neurons and normalized the desensitization of CAPS responses in FIC neurons. Our data suggest that FIC afferent neurons exhibit abnormal CAPS responses. The latter may be due to enhanced endogenous activities of PKC. PMID- 15882788 TI - Alterations in the substance P-induced anti-nociception in the central nervous system of rats after morphine tolerance. AB - The present study investigated the alteration in intracerebroventricular administration of substance P-induced anti-nociception after morphine tolerance in rats. The nociceptive response latencies to noxious thermal and mechanical stimulation increased significantly after intracerebroventricular administration of SP in normal rats. The SP-induced anti-nociception decreased significantly in morphine-tolerant rats than that in opioid naive rats. The results demonstrated that SP induced significant anti-nociceptive effects in the brain in normal rats, and the SP-induced anti-nociception was attenuated significantly in morphine tolerant rats than that in opioid naive rats, indicating an alteration in SP induced anti-nociception in the brain after chronic morphine treatments. PMID- 15882789 TI - Coordination of hand grip and load forces in uni- and bidirectional static force production tasks. AB - The purpose of the study was to explore the differences in coordination of grip (G) and load forces (L) in a unidirectional and bidirectional bimanual static force production task. Subjects (N=10) exerted oscillatory isometric L profiles against an externally fixed hand-held device, modulated either in pure tension (unidirectional) or in alternating tension and compression (bidirectional) at a rate of either 1.33 or 2.67 Hz. The unidirectional task revealed a high level of coordination of both the ipsilateral (i.e., G and L of each hand) and contralateral pairs of forces (two Gs and two Ls) as assessed by correlation and stability of force ratios. The bidirectional task demonstrated a low level of inconsistently modulated Gs with respect to the change of L, which resulted in a deteriorated coordination, particularly between the ipsilateral forces. The overall effect of task on the force coordination was higher than the effect of frequency suggesting that the higher frequency of G modulation required in the bidirectional task is not likely to be the main cause of the observed phenomenon. We interpret these differences by a relative simplicity of the control mechanisms of the unidirectional task based on a single synergy of G and L muscles that allows simultaneous coordination of both the ipsilateral and contralateral forces. Due to the switching between two distinctive synergies involving G muscles, the bidirectional task could possess a higher control complexity causing a decoupled coordination of the ipsilateral forces, while retaining the coordination of contralateral forces at a relatively high level. PMID- 15882790 TI - Expression analysis of psychological stress-associated genes in peripheral blood leukocytes. AB - In this study, we have developed a microarray including 1467 cDNAs that were selected to specifically measure stress response in peripheral blood leukocytes. Venous blood was collected from 10 graduate students 2 h before and 2 or 24 h after an open presentation for their Ph.D. The mRNA levels in leukocytes were compared with those prepared 4 weeks before the presentation. Hierarchical cluster showed that distinct groups of genes uniformly changed their expression values in response to the stress. Bayesian t test identified significantly up regulated 49 genes and down-regulated 21 genes. Most of them are categorized into cytokines, cytokine receptors, growth- or apoptosis-related molecules, and heat shock proteins, suggesting that stressful life events trigger acute responses in leukocytes. Our results suggest that gene expression profile in peripheral blood leukocytes may be a potentially useful method for the assessment of complex stress responses. PMID- 15882791 TI - Spatio-temporal dynamics of the visual system revealed in binocular rivalry. AB - From the evolutionary viewpoint, animals need to monitor the surrounding environment and capture salient features, such as motion, for survival. The visual system is highly developed for monitoring a wide area of visual field and capturing such salient features. In humans and primates, there is a wide binocular field, suggesting a necessity of integrating the images from the two eyes. Binocular rivalry [R. Blake, A neural theory of binocular rivalry, Psychol. Rev. 96 (1989) 145-167; R. Blake, N.K. Logothetis, Visual competition, Nat. Rev. Neurosci. 3 (2002) 13-21], where incompatible inputs from the two eyes compete to emerge in the subject's visual percept, has been shown to exhibit highly adaptive behavior [I. Kovacs, T.V. Parathomas, M. Yang, A. Feher, When the brain changes its mind: interocular grouping during binocular rivalry. Proc. Natl. Acad. Sci. U.S.A. 93 (1996) 15508-15511; N.K. Logothetis, Single units and conscious vision, Philos. Trans. R. Soc. Lond. B. Biol. Sci. 353 (1998) 1801-1818]. Here we investigated the spatio-temporal dynamics of the ocular dominance pattern in binocular rivalry under conditions where conflicting salient features were presented in a temporally varying manner. We found a striking example of the detailed structure of the dominance wave propagation, by using a spatio-temporal sampling method. The data show in detail the ability of the visual system to dynamically adapt to the changing stimuli in the context of the massively parallel visual field. We show by model prediction that the globally coherent dominance change in the presence of multiple stimuli can be explained by a mechanism based on local saliency comparison. PMID- 15882792 TI - Prognostic significance of apoptosis in medulloblastoma. AB - Since apoptosis is a major contributor to cell loss in medulloblastoma, either spontaneous or induced by radiation and chemotherapy, the apoptotic rate in resection specimens could have prognostic significance. We analysed the apoptotic rate in 58 medulloblastoma resection specimens using an antibody against cleaved caspase 3, a specific marker of apoptotic cell death, and tested its possible prognostic significance. The apoptotic rate varied considerably among medulloblastomas (0.1-25.9%, median 1.1%). Apoptotic cells were relatively evenly distributed in 39 cases, while in 19 cases, an uneven distribution with foci of an increased number of apoptotic cells and their clustering was observed. Clusters of apoptotic cells were found around necrotic areas, while necrotic cells were caspase 3 negative. The apoptotic rate was higher in medulloblastomas with CSF dissemination, tended to be higher in desmoplastic medulloblastomas, but there was no association with age group and sex. In the univariate analysis of overall survival, the apoptotic rate had no prognostic value. The variation in apoptotic rate among medulloblastomas is very likely predominantly associated with variations in tumour microenvironment, as supported by apoptotic cell clustering and rimming around necrotic areas. The apoptotic rate in medulloblastoma resection specimens does not seem to be of prognostic value. PMID- 15882793 TI - SUMO-1 marks subdomains within glial cytoplasmic inclusions of multiple system atrophy. AB - Conjugation of the small ubiquitin-like modifier, SUMO-1, to target proteins is linked to the regulation of multiple cellular pathways, including nucleocytoplasmic trafficking, cell cycle progression, the ubiquitin-proteasome system and apoptosis. Recently, the accumulation of SUMOylated proteins in pathological neuronal intranuclear aggregates has been found in several neurodegenerative diseases. The aim of our study was to examine SUMO-1 in the alpha-synucleinopathy diseases, Multiple System Atrophy (MSA) and Dementia with Lewy Bodies (DLB). We conducted anti-SUMO-1 immunostaining of fixed brain tissue sections and smears of unfixed brain tissue homogenates of DLB and MSA cases. We found that oligodendroglial cytoplasmic inclusions, the alpha-synuclein-positive cytoplasmic aggregates that characterize MSA, exhibit robust punctate SUMO-1 immunostaining, marking discrete submicron-sized subdomains within the inclusion bodies. Lewy bodies in smears of DLB tissue homogenates showed similar SUMO-1 positive structures, although these were not detected in fixed tissue. In cell culture experiments, we found that the nuclear and perinuclear accumulation of SUMO-1 aggregates could be induced in glioma cells by chemical inhibition of proteasomal protein degradation. PMID- 15882794 TI - Multiple kinase pathways mediate the early sciatic ligation-associated activation of CREB in the rat spinal dorsal horn. AB - Phosphorylation of the cyclic AMP response element-binding protein (CREB) in the spinal dorsal horn may critically contribute to chronic pain following peripheral nerve injury. We employed inhibitors and activators of protein kinase A (PKA), protein kinase C (PKC), extracellular signal-regulated kinase 1 and 2 (ERK1/2) and calcium/calmodulin-dependent kinase II (CaMKII) to examine whether these kinases individually or in concert mediate the increase in CREB phosphorylation that is evident as early as 2 h after loose ligation of the sciatic nerve. Specific inhibitors of each kinase significantly attenuated the ligation associated CREB phosphorylation when compared to saline-treated animals. Combined application of the ERK1/2 and CaMKII inhibitors also attenuated the ligation associated CREB activation but not to a greater extent than either inhibitor alone. Specific activators of PKA, PKC and ERK1/2 elicited significant increases in CREB phosphorylation 2 h after drug application in the spinal dorsal horn of control, peripherally uninjured animals. Pre-treatment of animals with the ERK1/2 inhibitor abolished the increases elicited by either the PKA or the PKC activator. Significant increases in ERK1/2 phosphorylation were also detected 2 h after sciatic ligation confirming a role for the ERK pathway in injury-related responses in the dorsal horn. Each kinase inhibitor significantly attenuated the ligation-associated activation of ERK1/2 as well. These data suggest that early, sciatic ligation-elicited phosphorylation of CREB in the spinal dorsal horn is mediated by multiple kinase pathways, and that PKA, PKC and CaMKII activate CREB at least in part by way of the ERK pathway. PMID- 15882795 TI - Aspirin modulates LPS-induced nitric oxide release in rat glial cells. AB - Nitric oxide and prostaglandins are among the numerous substances released by activated glial cells. The aim of this study was to evaluate the effect of high level aspirin on iNOS expression in cultured rat glial cells treated with lipopolysaccharide (LPS) as pathological stimulator. Using Western Blotting, we verified that aspirin enhanced LPS-induced iNOS expression and the presence of 15 deoxy-Delta(12,14)-prostaglandin (15d-PGJ2) suppressed this aspirin effect. However, the exposure of LPS-treated glial cells to aspirin resulted in a decrease of NO production. These results suggest that aspirin interferes with the cross-talk of prostaglandins and NO, blocking the endogenous negative control exerted by COX products on iNOS expression. On the other side, aspirin seems to act directly on iNOS reducing its activity, even if it does not completely block NO release by LPS-stimulated glial cells. Then aspirin could maintain homeostatic functions of NO, while it prevents toxic effects, corresponding to high NO concentrations. PMID- 15882796 TI - Anticipatory covariation of finger forces during self-paced and reaction time force production. AB - We tested a hypothesis that humans can change patterns of finger force covariation in a task-specific manner in preparation to a change in the total force. Subjects performed quick targeted force pulses by pressing with four fingers on force sensors from a certain background force level to a target level. In self-paced trials, finger force modes (hypothetical commands to fingers) showed changes in covariation, computed across trials, more than 100 ms before changes in the total force. Half of the subjects showed large early changes in force mode covariation, while in the other half these changes were much smaller and were followed by a larger positive covariation of finger modes potentially destabilizing the total force profile. Such early covariation changes were absent under the simple reaction time instruction. We conclude that anticipatory covariation reflects control processes that can be expressed differently in different persons and modified depending on the available time for action preparation. PMID- 15882797 TI - Stimulation of the nucleus locus coeruleus/subcoeruleus suppresses visceromotor responses to colorectal distention in the rat. AB - The aim of the present study was to examine whether the nucleus locus coeruleus/subcoeruleus (LC/SC) modulates visceromotor function. In the present study, an electromyogram (EMG) of the external abdominal oblique muscle evoked by colorectal distention was measured as a visceromotor reflex response, and inhibitory effects of LC/SC stimulation were estimated by the decrease of EMG activity. Under halothane anesthesia (1% in air), graded colorectal distentions (30, 60 or 80 mmHg) were produced by inflating a balloon inside the descending colon and rectum. A bipolar EMG electrode was inserted into the left external abdominal oblique muscle to record the EMG response to colorectal distention. Colorectal distention at a pressure of 30 mmHg did not evoke any EMG activity in the external abdominal oblique muscle in all rats tested. Electrical stimulation of the LC/SC (30, 50 and 70 microA, 100 Hz, 0.1 ms pulses) reduced EMG responses evoked by colorectal distention to 60 and 80 mmHg. LC/SC stimulation was effective both ipsilaterally and contralaterally indicating a bilateral effect. EMG responses decreased with an increase of LC/SC stimulation intensity. Following recordings of the inhibitory effects of LC/SC stimulation, lesions of the LC/SC ipsilateral to the EMG recording site were induced; 1 h after lesions the inhibitory effects of LC/SC stimulation were examined again. LC/SC stimulation did not reduce the EMG responses when LC/SC stimulation was applied to the ipsilateral LC/SC, whereas EMG responses were observed by stimulation of the intact LC/SC contralateral to the EMG recording site. From lesion experiments, it could be considered that suppression of the visceromotor response to colorectal distention is due to activation of the LC/SC. The results suggest that the visceromotor function is under the control of the centrifugal pathways from the LC/SC. PMID- 15882798 TI - Behavioral and affective changes in children and adolescents with chronic cerebellar lesions. AB - The aim of the present study was to investigate if clinically relevant affective or behavioral changes as described in adults in the cerebellar affective syndrome by Schmahmann and Sherman [The cerebellar cognitive affective syndrome, Brain 121 (1998) 561-579] are likely to occur as a long-term sequelae of cerebellar vermis lesions in children. Site and extent of the vermal lesion were defined on the basis of individual 3D-MRI scans and lesion data were correlated with behavioral and affective changes. Affect and behavior were assessed in children after cerebellar tumor surgery by means of experimenter ratings based on the description of the cerebellar affective syndrome and free ratings by the patients and their parents. Twelve children and adolescents with a former cerebellar astrocytoma surgery without subsequent radiation or chemotherapy participated. Detailed analysis of individual 3D-MR images revealed that lesions affected the vermis in nine children. Experimenter ratings according to Schmahmann revealed no relevant problems in patients. In five out of nine patients with vermal affection somewhat increased thoughtful, anxious or aggressive behavior was reported by patients and parents. In conclusion, minor behavioral and affective changes were present in a subset of children with chronic vermal lesions. PMID- 15882799 TI - The GABA type A receptor alpha5 subunit gene is associated with bipolar I disorder. AB - Several genetic studies have revealed that bipolar disorders are linked with the chromosomal locus of 15q11-q13, where the gamma-aminobutyric acid (GABA) receptor alpha5 subunit gene (GABRA5) locates. GABA is one of the major neurotransmitters that may be involved in the pathogenesis of bipolar disorder. Five polymorphisms in the GABRA5 gene, -754C>T in the promoter region, IVS1-21G>A, IVS2-26T>A, (*)302C>T in 3'-UTR of exon 5, and a CA repeat polymorphism in the 3' flanking region were examined in a Japanese population. IVS1-21G>A exhibited significant differences in the distribution of the genotype and allele frequency in bipolar I disorder patients but not in bipolar II disorder patients, compared with control subjects. The haplotype analysis showed that IVS1-21G>A/IVS2-26A>T was associated with bipolar I disorder, and the IVS1-21A/IVS2-26T haplotype was a negative risk factor for susceptibility to the disorders (odds ratio: 0.57, 95% confidence interval: 0.44-0.73). These results suggest that the GABRA5 gene may confer susceptibility to bipolar I disorder. PMID- 15882800 TI - Modulation of peripheral inflammation in sensory ganglia by nuclear factor (kappa)B decoy oligodeoxynucleotide: involvement of SRC kinase pathway. AB - Nuclear factor kappa B (NF(kappa)B) transcription factor plays a key role in the expression of many genes involved in the inflammatory process. We used the Freund's Complete Adjuvant (FCA)-induced model of peripheral inflammation to investigate the anti-inflammatory effects of double stranded oligodeoxynucleotides (ODN) with consensus NF(kappa)B sequence as transcription factor decoys to inhibit NF(kappa)kappaB activation in the dorsal root ganglia (DRG). Local administration of the wild-type-, but not mutant-ODN decoy, dose dependently inhibited edema formation and paw withdrawal latency as a measure of hyperalgesic response induced by FCA in rat paw. Biochemical assays performed in ipsilateral L4/L5 dorsal root ganglia obtained following FCA/wild-type ODN treatment showed: (1) an inhibition of the activity of c-Src kinase, a member of the non-receptor tyrosine kinase super family, (2) a decreased level of p65 NF(kappa)B subunit, and (3) an inhibition of cyclooxygenase-2 (COX-2) protein expression, a major pro-inflammatory enzyme transcriptionally controlled by NF(kappa)B. The present results indicate that the wild-type ODN decoy may act as a competitor for NF(kappa)B binding to its cognate recognition sequence as well as a modulator of c-Src activity in the DRG. The NF(kappa)B/c-Src interaction may represent a novel pathway for further exploring the molecular mechanism of inflammatory pain. PMID- 15882801 TI - The role of CGRP and nicotinic receptors in centrally evoked facial blood flow changes. AB - The release of CGRP in humans is associated with the occurrence of migraine headaches. The vasoactive neuropeptide is released by afferent neurones originating in the peripherally located trigeminal ganglion supplying the dura mater. The role of CGRP in migraine is further supported by recently released data showing that the CGRP-antagonist BIBN4096BS is clinically effective for the treatment of migraine headaches. Yet, the trigger for CGRP release during migraine attacks is not identified. It is suggested that the peripheral CGRP release during a migraine attack might be either triggered by direct activation of afferent dural neurones, or, by indirect activation via the central nervous system. Recently, we were able to show that the CGRP-antagonist BIBN4096BS is able to inhibit vasodilation induced by trigeminal ganglion stimulation. Now, we extend our studies to the investigation of facial blood flow changes induced by electrical stimulation of the brainstem trigeminal nucleus caudalis (TNC). Here, we show that stimulation of the TNC leads to a pronounced increase of facial blood flow. The nicotinic antagonist Hexamethonium reduced the evoked flow by approximately 50% (30 mg/kg), while the muscarinic antagonist Atropin did not influence the stimulation evoked blood flow. Application of BIBN4096BS (0.3 mg/kg, i.v.) diminished the evoked flow almost completely. Therefore, we conclude that CGRP represents the key player in TNC-induced facial vasodilation, while activation of nicotinic receptors modulates centrally induced peripheral neurogenic vasodilation. PMID- 15882802 TI - Preliminary analysis of the GABA-induced current in cultured CNS neurons of the cutworm moth, Spodoptera litura. AB - Properties of GABA-induced current in cultured CNS (ganglion) neurons of cutworm moths (Spodoptera litura) were studied using a whole-cell patch-clamp technique. CNS neurons ranging from 10 to 20 microm in diameter were cultured for 4-7 days in MGM-464 medium. GABA-induced a current response in these neurons in a sigmoidally dose dependent manner where the Hill coefficient and EC50 were 2.2 and 33.0 microM, respectively. The reversal potential of GABA-induced current was -2.5 mV, which is close to the Cl- equilibrium potential that was calculated from chloride ion concentrations in the present experimental environment. Furthermore, the GABA-induced current response depended on the extracellular chloride ion concentration, indicating that the receptor regulates the Cl- permeability of cells. The GABA-induced current was completely inhibited by the GABA(A) antagonist, SR95531, and activated by the GABA(A) agonist, muscimol but not by the GABA(B) agonist, baclofen. On the other hand, the GABA(C) agonist, CACA, also induced a little smaller current than the GABA-induced response. The pharmacological behaviors of the GABA-induced currents suggest that these cells contain GABA receptors that belong to the GABA receptor family including the Rdl GABA receptors identified in Drosophila neurons. PMID- 15882803 TI - UCHL1 is associated with Parkinson's disease: a case-unaffected sibling and case unrelated control study. AB - To avoid the possible confounding effect of population stratification, we employed a discordant sibling study design and a liberalization of the sibling transmission disequilibrium test to confirm the association of the S18Y variant of the ubiquitin carboxi-terminal hydrolase L1 (UCHL1) gene with Parkinson's disease (PD). The study included 497 case-control pairs (427 case-unaffected sibling pairs and 70 case-unrelated control pairs). Analyses confirmed a significant inverse association of the UCHL1 S18Y polymorphism with PD overall (OR=0.18, 95% CI=0.05-0.64, p=0.002, recessive model) and in several strata. PMID- 15882804 TI - Area postrema-lesions increase operant responding to sucrose in rats. AB - Rats with lesions of the area postrema (APX) are known to exhibit an enhanced intake of highly palatable foods such as sweetened condensed milk and cookies. These observations suggest the possibility that APX rats find these foods more rewarding and will work harder to obtain these foods. Sham and APX rats were tested on fixed ratio (FR) and progressive ratio (PR) schedules. APX rats consistently pressed more times to receive sucrose solution and attained both FR 3 and FR 5 criteria significantly faster than sham-lesioned control rats. Furthermore, rats with APX had significantly higher break points than sham lesioned control rats on a progressive ratio schedule. These results support the hypothesis that rats with lesions of the area postrema will consistently work harder to obtain a highly palatable food reward. PMID- 15882805 TI - Different chloride electrochemical gradients across the plasma membrane in subcellular compartments of rat cerebellum granules. AB - The effects of GABA on intracellular Ca2+ have been studied in neonatal rat cerebellum granule cells (CGC) in culture by Oregon Green and two-photon excitation fluorescence microscopy. This technique allowed the study of [Ca2+]i both in cell bodies and neurites. Working with a perfusion chloride concentration corresponding to the average extracellular level, we found that GABA induced an increase in [Ca2+]i in the cell bodies in many of the cells studied with a maximum at day 4 in vitro. This effect disappeared after day 6. However, no increase in [Ca2+]i was ever found in neurites at standard [Cl-]e. On the other hand, an increase of [Ca2+]i was found also in neurites when [Cl-]e was close to zero. The [Ca2+]i increases were blocked by both bicuculline methiodide and nimodipine. The results indicate the presence of an outward directed electrochemical gradient for chloride in the cell bodies which results in depolarization by GABA via GABA(A) receptor activation. Calcium ion influx ensues due to activation of voltage-gated calcium channels (VGCC). This phenomenon may mediate the well-known trophic effect of GABA on these cells at this developmental stage, via an action of [Ca2+]i on the transcriptional activity of the nucleus. No calcium accumulation takes place in neurites due to either no or a reverse (hyperpolarizing) electrochemical gradient for chloride ions. Such a circumstance in later developmental stages may be of importance for the phasic component of GABA-mediated inhibition. PMID- 15882806 TI - Altered brain stem responsivity to duodenal pain after a single stressful experience. AB - A single session of foot shock stress produces stable and long lasting sensitization of behavioral, hormonal and intestinal motility responses to novel stressful stimuli in laboratory rats. This is reflected in increased expression of the activity marker protein Fos in brain areas involved, following an external stressor. We present data from awake, freely moving rats in which a silicone balloon was surgically implanted in the duodenum. Firstly, cardiovascular reflexes to distentions were studied using telemetry with surgically implanted transmitters, 2 weeks after a single, 15-min session of foot shocks. The distentions caused characteristic, bi-phasic responses in both mean arterial blood pressure and heart rate that were not different between preshocked and control animals. Secondly, the numbers of Fos immunopositive cells were quantified in selected brain areas, 1 h after repeated distention of the duodenum. We found an increase in distention-induced Fos in preshocked rats in the nucleus tractus solitarius and a weaker effect in the central nucleus of the amygdala. This could be a first indication that altered visceral afferent processing in previously stressed rats, found earlier for the colon, may be a general and not an organ-specific phenomenon. PMID- 15882807 TI - Involvement of ryanodine receptors in IP3-mediated calcium signalling in neurons. A modelling approach. AB - Phospholipase C (PLC)-coupled metabotropic receptors trigger the release of intracellular Ca2+ through activation of IP3 receptors (IP3Rs). Increasing evidence suggests that they can also and perhaps more efficiently mobilize Ca2+ through ryanodine receptors (RyRs). We constructed a model allowing a variable PLC stimulation level (via the parameter gamma) as well as a variable involvement of RyRs (via the parameter A). The sole presence of RyRs (A not = 0) affected the basal Ca2+ concentration [Ca2+]i. To keep Ca2+ below 160 nM, we fixed the upper limit of A to 0.2, a value that is compatible with the numerical ratio between RyRs and IP3Rs in cerebellar Purkinje neurons. Metabotropic responses were simulated by abruptly raising the value of gamma to various levels. In the absence of RyRs, the model starts to oscillate with gamma=0.4. For lower levels of PLC stimulation (gamma< or =0.3), the presence of RyR is capable of triggering an oscillatory signal. When A< or =0.18, the frequency of the Ca2+ oscillations augments from 0.1 to 1.3 Hz as a function of gamma. Conversely, as the frequency increases, the amplitude of the oscillations is reduced from 1 microM to 50 nM. With higher values of A, the oscillating pattern is definitively inhibited. It is concluded that RyRs have the potentiality to strikingly affect the temporal pattern of the Ca2+ signalling triggered by IP3-related metabotropic responses. PMID- 15882808 TI - Estradiol regulates APP mRNA alternative splicing in the mice brain cortex. AB - Alternative splicing of amyloid precursor protein (APP), one of the candidate genes for Alzheimer's disease, yields three major mRNAs, which give rise to APP770, APP751 and APP695 protein isoforms. Out of these three isoforms, APP695 is expressed most predominantly in the brain. The splicing and processing of APP are shown to be influenced by several factors including hormones. In this study, we report the effect of withdrawal and administration of sex steroid hormones on the alternative splicing of APP mRNA during aging in the brain cortex of mice of both sexes. The level of APP695 mRNA isoform was higher in intact adult as compared to old mice of both sexes. Gonadectomy upregulated the APP695 mRNA isoform levels in all groups except in adult female where the level was downregulated. Estradiol supplementation upregulated the level of APP695 mRNA isoform in all groups except in old male where the level was downregulated. Thus these results show that the level of APP695 mRNA changes with age and estradiol may play a key role in the development of Alzheimer's disease by modulating the level of APP mRNA isoforms. PMID- 15882809 TI - Rapid tolerance to focal cerebral ischemia in rats is induced by preconditioning with electroacupuncture: window of protection and the role of adenosine. AB - The aim of the present study was to investigate the first protective window of preconditioning with electroacupuncture (EA) against focal cerebral ischemia, and to explore whether adenosine is involved in the rapid tolerance phenomenon. Sixty four male Sprague-Dawley rats were randomly assigned to eight groups (n=8 in each). Animals in the control group received no treatment, and animals in EA1-EA4 groups received EA at 0.5, 1, 2 and 3 h before induction of focal cerebral ischemia, respectively. Rats in DPCPX group were intraperitoneally injected with 1 mg kg-1 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), 3 h before induction of focal cerebral ischemia. Animals in vehicle group and EA+DPCPX group were pretreated with 1 ml kg-1 dimethyl sulfoxide (DMSO, the solvent of DPCPX) and 1 mg kg-1 DPCPX 30 min before preconditioning with EA, respectively. All rats were anesthetized with 40 mg kg-1 pentobarbital sodium intraperitoneally. Animals that required EA preconditioning, received EA with intensity of 1 mA and frequency of 15 Hz at the Baihui acupoint (GV 20) for 30 min. The focal cerebral ischemia was produced by the right middle cerebral artery occlusion (MCAO) for 120 min. The neurologic deficit scores (NDS) and brain infarct volumes were evaluated at 24 h after reperfusion. All rats survived until 24 h after reperfusion. Preconditioning with EA at 2 h before induction of focal cerebral ischemia improved neurologic outcome (P<0.05 versus control) and reduced the infarct volume (P<0.01 versus control) at 24 h after reperfusion. These beneficial effects were reversed by pretreatment with 1 mg kg-1 DPCPX, whereas this agent itself did not affect the NDS and volume in drug-ischemic controls after ischemia. The results show that preconditioning with single EA session induces rapid tolerance to focal cerebral ischemia. The rapid ischemic tolerance appears at 2 h (but not at 0.5, 1, or 3 h) after preconditioning, and is possibly mediated through an adenosine A1 receptor-related mechanism. PMID- 15882810 TI - Chronic ethanol modulates delta and mu-opioid receptor expression in rat CNS: immunohistochemical analysis with quantitiative confocal microscopy. AB - Ethanol consumption affects levels of endogenous opioids as well as opioid receptors in both animals and humans. We studied the expression of delta (delta) and mu (mu) opioid receptors (ORs) in brain sections of adult male Sprague-Dawley rats after 2 weeks of consuming ethanol in a liquid diet, with comparisons to sections from pair-fed control animals. Immunohistochemical staining for the ORs, using selective antibodies, and quantitation of confocal images, revealed increased expression of delta-ORs in hippocampal CA1 of the chronic ethanol treated rats. In contrast, mu-ORs decreased in their expression after ethanol treatment in multiple brain areas, including cortex, hippocampus, midbrain colliculi, striatum and nucleus accumbens. The alterations in immunoreactive OR expression may be related to reduced functional coupling of the ORs to G proteins, as found in prior studies in several brain regions, using the same chronic ethanol diet protocol. Changes in OR expression and functional coupling in the CNS may be factors in ongoing ethanol consumption and tolerance. PMID- 15882811 TI - Minor expression of fascin-1 gene (FSCN1) in NTera2 cells depleted of CREB binding protein. AB - CREB-binding protein (CBP) is a transcriptional coactivator whose mutations may cause a generalized perturbation of gene expression. We silenced the CBP gene in NT2 neuronal precursor cells by RNA interference. Hybridization experiments on 1.2K human cDNA microarrays showed that the FSCN1 gene, encoding for fascin-1 protein, was clearly less expressed in CBP-depleted cells than in controls. This reduction was confirmed by Real Time PCR and Western blotting assays. We also analyzed FSCN1 expression profile during NT2 neuronal differentiation induced by retinoic acid (RA), showing that FSCN1 was up-regulated during neurogenesis. This mRNA increasing suggests the importance of fascin-1 in the formation of mature neurons, in accordance with its actin-bundling function and its localization in neurites and neuronal growth cones. The lower amount of FSCN1 transcript in the absence of the CBP factor was also established in RA-treated cells. In conclusion, this research supports FSCN1 as a novel marker of NT2 neuronal differentiation and the possible role of CBP in its regulation. PMID- 15882812 TI - Lipooligosaccharide of Campylobacter jejuni prevents myelin-specific enteral tolerance to autoimmune neuritis--a potential mechanism in Guillain-Barre syndrome? AB - Campylobacter jejuni-induced enteritis is the most common infection preceding Guillain-Barre syndrome (GBS), an immune-mediated polyradiculoneuritis. The acute autoimmune attack is thought to be based on C. jejuni antigens which may mimick antigens of the peripheral nervous system. Additional pathomechanisms, like disturbance of natural T cell immunoregulation by C. jejuni, have not been evaluated so far. In experimental autoimmune neuritis (EAN), a T lymphocyte mediated animal model of human GBS, tolerance to myelin-derived autoantigens can be induced by oral feeding of the respective antigen. Here we investigated whether the lipooligosaccharide (LOS) fraction of C. jejuni may directly alter immunologic tolerance through gastrointestinal pathways. While EAN, actively induced by immunization with bovine peripheral nerve myelin could be ameliorated by precedent feeding of myelin, feeding of C. jejuni LOS along with the myelin antigen not only prevented the tolerizing effects of oral myelin but even accelerated the onset of overt EAN and augmented the myelin-specific B cell response. These findings provide evidence that LOS of C. jejuni, as produced in the gut during C. jejuni-induced enteritis, can disturb natural tolerance to definite proteins which may be or may mimic peripheral nerve antigens. In human patients this may be one of the potential mechanisms to explain why C. jejuni enteritis is a common trigger of GBS. PMID- 15882813 TI - A new fast electrophysiological response measured extracellularly in a mouse hippocampal brain slice. AB - In a hippocampal mouse brain slice the CA1 stratum radiatum field potential evoked by electrical stimulation of the Schaffer collaterals was used to analyze the very early responses. Parallel bipolar electrodes were utilized to lower the voltage induced by the stimulating current in the recording electrode, and to keep the induced voltage within the linear range of the headstage preamplifier and the recording amplifiers. When the stimulating and recording electrodes are in saline in the same plane and same depth, the shapes of the recorded voltage and stimulating current are similar. The shape of the recorded voltage significantly changes when the recording electrode is in the stratum radiatum of CA1. The signal recorded in the first 2 ms, before the presynaptic volley, indicates the existence of a fast response shown as a positive deflection. For times slightly longer than the stimulating pulse width, the fast response overlaps with the voltage induced by the discharging of the polarization charge accumulated in the double layer capacitance at the interface between metal and saline. For times longer than about 1.4 ms the fast response and presynaptic volley are superimposed. The fast response is about 0.5 mV when the stimulating charge is 15-25 nC. The demonstration that the fast response disappears upon bath perfusion of tetrodotoxin is convincing support for its axonal origin. We postulate that this fast response represents the current source corresponding to the Schaffer collaterals excitatory sink. An undistorted recording of the first 2 ms of the sweep gives useful information about the fast neuronal activity measured extracellularly, and cannot be regarded as a simple stimulus artifact. PMID- 15882814 TI - Evidence for the involvement of glutamatergic system in the antinociceptive effect of ascorbic acid. AB - This study examined the role of glutamatergic system in the ascorbic acid (AA) induced antinociception in chemical behavioural models of nociception in mice. AA (0.3-10 mg/kg, i.p.) produced significant inhibition of both phases of formalin induced licking, with mean ID50 values of 4.0 and 3.2 mg/kg and inhibitions of 56+/-4 and 60+/-7% for the early and second phase of the nociception caused by formalin, respectively. AA (1-5 mg/kg, i.p.) also produced significant inhibition of glutamate-induced nociception with mean ID50 value of 2.1 mg/kg and inhibition of 66+/-5%. Furthermore, AA (3 mg/kg, i.p.) caused marked inhibition of nociceptive response induced by intrathecal injection of glutamate, NMDA, AMPA, kainate and substance P, with inhibitions of 49+/-9, 42+/-7, 34+/-8, 38+/-5 and 65+/-8%, respectively. In contrast, AA at the same dose did not affect the biting response induced by the metabotropic agonist trans-ACPD. Taken together, present results indicate that AA, at low systemic doses, produces a rapid onset and consistent antinociception in mice when assessed in several models of chemical nociception, an action that is likely mediated by an interaction with ionotropic, but not metabotropic, glutamate receptors. PMID- 15882815 TI - Role of tissue plasminogen activator/plasmin cascade in delayed neuronal death after transient forebrain ischemia. AB - We studied the possible involvement of the tissue plasminogen activator (t PA)/plasmin system on both delayed neuronal death in the hippocampus and the associated enhancement of locomotor activity in rats, after transient forebrain ischemia induced by a four-vessel occlusion (FVO). Seven days after FVO, locomotor activity was abnormally increased and, after 10 days, pyramidal cells were degraded in the CA1 region of the hippocampus. FVO increased the t-PA antigen level and its activity in the hippocampus, which peaked at 4 h. Both the enhanced locomotor activity and the degradation of pyramidal cells were significantly suppressed by intracerebroventricular injection of aprotinin, a plasmin inhibitor, at 4 h but not during FVO. These results suggest the importance of the t-PA/plasmin cascade during the early pathological stages of delayed neuronal death in the hippocampus following transient forebrain ischemia. PMID- 15882816 TI - Activation of spinal extracellular signaling-regulated kinases by intraplantar melittin injection. AB - Intraplantar injection of melittin, a major toxic peptide of whole bee venom, has been proved to cause alteration in both behavioral and spinal neuronal responses in rats. To see whether extracellular signaling-regulated kinases (ERK) in the spinal cord dorsal horn are activated and involved in induction and maintenance of persistent ongoing nociception, pain hypersensitivity and inflammation, three doses of U0126 (1,4-diamino-2,3-dicyano-1, 4-bis-[o aminophenylmercapto]butadiene), a widely used specific MAP kinase kinase (MEK) inhibitor, were administered through chronic intrathecal catheterization prior to or after intraplantar injection of melittin. We found that: (1) the induction of melittin-induced persistent spontaneous nociception (PSN), mechanical and heat hypersensitivity could be suppressed by U0126 in a dose-related manner; (2) specific inhibition of ERK pathway suppressed the maintenance of melittin-induced PSN and heat hypersensitivity, while established mechanical hypersensitivity could not be reversed; and (3) intrathecal administration of U0126 had no effects on peripheral inflammation induced by melittin. This result suggests that spinal ERK pathway might be a common factor involved in inducing and maintaining pathophysiological processes of ongoing pain and heat hyperalgesia, while the role of ERK pathway in generation of the mechanical hypersensitivity is not consistent and remains to be further clarified. PMID- 15882817 TI - Noise-induced cell death in the mouse medial geniculate body and primary auditory cortex. AB - Noise-induced effects within the inner ear have been well investigated for several years. However, this peripheral damage cannot fully explain the audiological symptoms in noise-induced hearing loss (NIHL), e.g. tinnitus, recruitment, reduced speech intelligibility, hyperacusis. There are few reports on central noise effects. Noise can induce an apoptosis of neuronal tissue within the lower auditory pathway. Higher auditory structures (e.g. medial geniculate body, auditory cortex) are characterized by metabolic changes after noise exposure. However, little is known about the microstructural changes of the higher auditory pathway after noise exposure. The present paper was therefore aimed at investigating the cell density in the medial geniculate body (MGB) and the primary auditory cortex (AI) after noise exposure. Normal hearing mice were exposed to noise (10 kHz center frequency at 115 dB SPL for 3 h) at the age of 21 days under anesthesia (Ketamin/Rompun, 10:1). After 1 week, auditory brainstem response recordings (ABR) were performed in noise exposed and normal hearing animals. After fixation, the brain was microdissected and stained (Kluever Barrera). The cell density in the MGB subdivisions and the AI were determined by counting the cells within a grid. Noise-exposed animals showed a significant ABR threshold shift over the whole frequency range. Cell density was significantly reduced in all subdivisions of the MGB and in layers IV-VI of AI. The present findings demonstrate a significant noise-induced change of the neuronal cytoarchitecture in central key areas of auditory processing. These changes could contribute to the complex psychoacoustic symptoms after NIHL. PMID- 15882818 TI - Comparing the accuracy of video-oculography and the scleral search coil system in human eye movement analysis. AB - The measurement of eye movements in three dimensions is an important tool to investigate the human vestibular and oculomotor system. The primary methods for three dimensional eye movement measurement are the scleral search coil system (SSCS) and video-oculography (VOG). In the present study, we compare the accuracy of VOG with that of SSCS using an artificial eye. We then analyzed the Y (pitch) and Z (yaw) component of human eye movements during saccades, smooth pursuit and optokinetic nystagmus, and the X (roll) component of human eye movement during the torsional vestibulo-ocular reflex induced by rotation in normal subjects, using simultaneous VOG and SSCS measures. The coefficients of the linear relationship between the angle of a simulated eyeball and the angle measured by both VOG and SSCS was almost unity with y-intercepts close to zero for torsional (X), vertical (Y) and horizontal (Z) movements, indicating that the in vitro accuracy of VOG was similar to that of SSCS. The average difference between VOG and SSCS was 0.56 degrees , 0.78 degrees and 0.18 degrees for the X, Y and Z components of human eye movements, respectively. Both the in vitro and in vivo comparisons demonstrate that VOG has accuracy comparable to SSCS, and is a reliable method for measurement of three dimensions (3D) human eye movements. PMID- 15882819 TI - Argon laser assisted small fenestra stapedotomy for otosclerosis. AB - To report and analyse our results and complications of argon laser assisted stapedotomy for primary otosclerosis. A retrospective analysis of 135 consecutive cases of primary otosclerosis operated by the senior author (JH) has been performed. The air-bone gap was calculated by using the pure tone average at 500, 1000, 2000 and 4000 Hz. A separate analysis of air-bone gap at 500, 1000 and 2000 Hz was carried out to assess the effects of a small diameter piston on low frequency hearing. Hearing at high frequencies (4000 and 8000 Hz) was also assessed to evaluate effects of small fenestra technique on high frequency hearing. Preservation of cochlear function was assessed by comparing the average pre- and post-operative bone conduction thresholds. Complications arising were analysed. The post-operative air-bone gap at 0.5, 1 and 2 kHz was 10 dB or less in 85.19% of patients and 20 dB or less in 97.04% patients. The air-bone gap at 0.5-4 kHz was 2)-alpha-L-Araf-(1-->3)-[alpha-L-Araf-(1-->4)]-beta-D-Galp-(1-->. This structure does not fit into any of the known classes of arabinogalactans. SEC/MALS experiments gave a molecular mass for the arabinogalactan isolated as 47 +/- 4 kDa but the original structure was probably larger. PMID- 15882848 TI - The structure of the antigenic O-polysaccharide of the lipopolysaccharide of Edwardsiella ictaluri strain MT104. AB - The structural characterization of the antigenic O-polysaccharide component of the lipopolysaccharide produced by the fish pathogenic bacterium Edwardsiella ictaluri MT104 was undertaken by the application of NMR spectroscopy and chemical analysis. The O-chain was found to be a linear polymer of a repeating tetrasaccharide unit composed of D-glucose, 2-acetamido-2-deoxy-D-galactose, and D-galactose in a 1:2:1 ratio having the structure: [carbohydrate structure]; see text. PMID- 15882849 TI - Oxygen solubility and permeability of carbohydrates. AB - The saturated oxygen concentration in a series of aqueous solutions of sorbitol (up to 35% w/w) and maltitol (up to 50% w/w) was measured using colorimetric reagent vials based on Rhodazine D. The results indicate that the solubility of oxygen in low-water carbohydrates is considerably lower than its solubility in pure water. It was concluded that the low-oxygen solubility is a major factor contributing to the barrier properties of low-water content carbohydrates used in the encapsulation of flavours, lipids, peptides and other oxidisable species. PMID- 15882850 TI - Molecular dynamics and atomic charge calculations in the study of heparin conformation in aqueous solution. AB - HF/6-31G** and molecular dynamics (MD) simulations were used to evaluate the performance of different atomic charge basis sets (i.e., Mulliken, Lowdin, and Electrostatic Potential Derived Charges--ESP) in heparin simulations. HF/3-21 G calculations were also used to study the NMR conformation of the IdoA residue. The results thus obtained indicated that ESP and Lowdin charges gave the better results in heparin simulations, followed by Mulliken charges, and that the minimum-energy conformation of IdoA can be different from that observed by NMR spectroscopy by less than 1 Angstrom. However, it was found that this small conformational modification is capable of inducing a change of almost 200 kJ/mol in the interactions of heparin with the surrounding environment, which is a meaningful amount of energy in the context of ligand-receptor interactions. This information can be potentially of great relevance in the design of heparin derived antithrombotic compounds. PMID- 15882851 TI - Syntheses of chlorogenin 6alpha-O-acyl-3-O-beta-chacotriosides and their antitumor activities. AB - Chlorogenin 3-O-beta-chacotrioside (1) and its 6alpha-O-acyl derivatives (2-6) were concisely synthesized. Introduction of a hydroxyl or acyloxy group onto the C-6 of the steroidal aglycone of dioscin decreased significantly the cytotoxicity of the parent saponin (dioscin). PMID- 15882852 TI - Evidence of two-step deprotonation of D-mannitol in aqueous solution. AB - Deprotonation of D-mannitol was studied in aqueous basic solutions by means of potentiometry and (13)C NMR spectroscopy. Two-step dissociation in the pH range from 12 to 13.8 was shown, and successive dissociation constants K(a1) and K(a2) were determined. In a solution with ionic strength I = 1.0 M (NaOH + NaNO(3)) pK(a1) = 13.1 +/- 0.1 and pK(a2) = 13.8 +/- 0.2. With increasing ionic strength from 0.75 to 3.0 M, both pK(a1) and pK(a2) values decrease. Deprotonation-induced chemical shifts in pH-variable (13)C NMR spectra show that the OH-groups next to internal carbon atoms C-3 and C-4 dissociate to a greater extent compared to OH groups next to external carbon atoms C-1 and C-6. PMID- 15882853 TI - New cyclomaltoheptaose (beta-cyclodextrin) derivative 2-O-(2 hydroxybutyl)cyclomaltoheptaose: preparation and its application for the separation of enantiomers of drugs by capillary electrophoresis. AB - A new soluble cyclomaltoheptaose (cyclodextrin) derivative, 2-O-(2 hydroxybutyl)cyclomaltoheptaose [2-O-(2-hydroxybutyl)-beta-cyclodextrin, 2-HB beta-CD], was prepared and studied as an efficient chiral selector in the separation of racemic mixtures of drugs by capillary electrophoresis (CE). Results showed that 2-HB-beta-CD could provide higher separating capability than that of beta-CD and the similarly substituted 2-HP-beta-CD. PMID- 15882854 TI - Correlation between antitumor activity, molecular weight, and conformation of lentinan. AB - A (1-->3)-beta-D-glucan having (1-->6) branching (L-FV-IB) from Lentinus edodes in water was degraded into seven fractions of different molecular weights by ultrasonic irradiation, and each was further fractionated into three parts, by precipitation from water into acetone at room temperature. The weight-average molecular weight (M(w)), radius of gyration ((z)(1/2)), and intrinsic viscosity ([eta]) of lentinan and its fractions in 0.9% NaCl aqueous solution and dimethyl sulfoxide (Me(2)SO) were determined by size-exclusion chromatography combined with multi-angle laser light scattering (SEC-LLS), LLS, and viscometry. Analysis of M(w), [eta], and (z)(1/2) in terms of known theory for worm like chains yielded 2240 +/- 100 nm(-1), and 100 +/- 10 nm for molar mass per unit contour length (M(L)), and persistence length (q), respectively, corresponding with theoretical data for triple-helical chains. The [alpha](D) of lentinan in water-Me(2)SO mixtures indicated an order-disorder transition. The results indicated that lentinan exists as a triple helix in 0.9% NaCl aqueous solution and as a single flexible chain in Me(2)SO. Assays in vivo and in vitro against the growth of Sarcoma 180 solid tumor as well as the colorimetric 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method for lentinan showed that the triple-helix sample exhibited a relatively high inhibition ratio. Interestingly, the triple-helix lentinan with M(w) of 1.49 x 10(6) exhibited the highest antitumor activity in vivo, having an inhibition ratio (xi) of 49.5%, close to that of 5-fluorouracil (xi = 50.5%), whereas the bioactivity (xi = 12.3%) of its single flexible chains almost disappeared. The triple-helix conformation plays an important role in enhancing the antitumor effects of lentinan. PMID- 15882855 TI - Adamantoylated monosaccharides: new compounds for modification of the properties of cyclodextrin-containing materials. AB - Adamantoyl glycosides were obtained in good yields by coupling adamantanecarboxylic acid with monosaccharides. They form very stable inclusion complexes with beta-cyclodextrin, as shown by (1)H NMR measurements. PMID- 15882856 TI - An enzymatically produced novel cyclic tetrasaccharide, cyclo-{-->6)-alpha-D-Glcp (1-->4)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->4)-alpha-D-Glcp-(1-->} (cyclic maltosyl-(1-->6)-maltose), from starch. AB - A bacterial strain M6, isolated from soil and identified as Arthrobacter globiformis, produced a novel nonreducing oligosaccharide. The nonreducing oligosaccharide was produced from starch using a culture supernatant of the strain as enzyme preparation. The oligosaccharide was purified as a crystal preparation after alkaline treatment and deionization of the reaction mixture. The structure of the oligosaccharide was determined by methylation analysis, mass spectrometry, and (1)H and (13)C NMR spectroscopy, and it was demonstrated that the oligosaccharide had a cyclic structure consisting of four glucose residues joined by alternate alpha-(1-->4)- and alpha-(1-->6)-linkages. The cyclic tetrasaccharide, cyclo-{-->6)-alpha-D-Glcp(1-->4)-alpha-D-Glcp(1-->6)-alpha-D Glcp(1-->4)-alpha-D-Glcp(1-->}, was found to be a novel oligosaccharide, and was tentatively called cyclic maltosyl-maltose (CMM). CMM was not hydrolyzed by various amylases, such as alpha-amylase, beta-amylase, glucoamylase, isoamylase, pullulanase, maltogenic alpha-amylase, and alpha-glucosidase, but hydrolyzed by isomalto-dextranase to give rise to isomaltose. This is the first report of the cyclic tetrasaccharide, which has alternate alpha-(1-->4)- and alpha-(1-->6) glucosidic linkages. PMID- 15882857 TI - Structure of the O-specific polysaccharide from Shewanella japonica type strain KMM 3299T containing the rare amino sugar Fuc4NAc. AB - An acidic O-specific polysaccharide (PS) of the agar-digesting bacterium Shewanella japonica with the type strain KMM 3299(T) was obtained by mild acid hydrolysis of the lipopolysaccharide. The polysaccharide was studied by component analysis, methylation analysis, (1)H and (13)C NMR spectroscopy, including 2D NMR experiments. The PS was determined to have the following structure involving three unusual amino sugars: PMID- 15882858 TI - Structure of an acidic polysaccharide from the marine bacterium Pseudoalteromonas aliena type strain KMM 3562T containing two residues of L-serine in the repeating unit. AB - The structure of an acidic polysaccharide from Pseudoalteromonas aliena type strain KMM 3562(T) has been elucidated. The polysaccharide was studied by component analysis, (1)H and (13)C NMR spectroscopy, including 2D NMR experiments. A (1)H, (13)C band-selective constant-time heteronuclear multiple bond connectivity experiment was used to determine amide linkages, between serine and uronic acid (UA) residues, via (3)J(H,C) correlations between Ser-alphaH and UA-C-6. It was found that the polysaccharide consists of pentasaccharide repeating units with the following structure: [carbohydrate structure]; see text. PMID- 15882859 TI - Pregnancy zone protein is a carrier and modulator of placental protein-14 in T cell growth and cytokine production. AB - A successful pregnancy can only occur when the maternal immune system fails to attack the allogeneic fetus. Two plasma proteins with described immunoregulatory activities, pregnancy zone protein (PZP) and placental protein-14 (PP14; also known as glycodelin-A), increase dramatically during pregnancy, prompting us to examine their potential role in mediating fetal protection. First, we demonstrated that both native PZP and its receptor-recognized monoamine-activated form (MA-PZP) bound non-covalently and specifically to PP14, exhibiting K(d) values greater than 3 microM, as determined by surface plasmon resonance. Our evidence further suggests that PZP is potentially a more effective carrier of PP14 than its relative alpha2-macroglobulin. Second, we found that T-cell activation, as measured by increased proliferation and IL-2 production, was inhibited by either PZP or PP14 in a dose-dependent manner. However, when PZP and PP14 were combined, they acted synergistically to inhibit T cell proliferation and IL-2 production. Interestingly, the combination of PZP and PP14 had little effect on the production of T(H)2 cytokine, IL-4. Based upon these findings, we hypothesize that PZP and PP14 form a stable complex in the plasma of pregnant women and together act synergistically to selectively modulate T-cell activation. Mechanistically, this activity appears to be independent of the PZP receptor (CD91) or PZP's anti-proteinase activity. PMID- 15882860 TI - Effect of ischemic preconditioning on reactive oxygen species-mediated ischemia- reperfusion injury in the isolated perfused rat lung. AB - OBJECTIVE: To investigate the protective effect of ischemic preconditioning (IP) in the early phase of reperfusion injury. DESIGN AND METHODS: Control rat lungs were subjected to 3 h of perfusion, whereas the lungs of the ischemia-then reperfusion (I/R) group were subjected to 2 h of cold ischemia following 30 min of perfusion. IP was performed by two cycles of 5-min ischemia followed by 5 min of reperfusion prior to 2-h cold ischemia. Lipid peroxidation and antioxidant status were determined in tissue samples. RESULTS: Lipid peroxidation and reduced/oxidized glutathione (GSH/GSSG) ratio were increased; antioxidant enzyme activities were decreased in the I/R group whereas lipid peroxidation and GSH/GSSG ratio were decreased; antioxidant enzyme activities were increased in the IP group. CONCLUSION: IP appeared to have a protective effect against reactive oxygen species-mediated I/R injury in isolated rat lung. PMID- 15882861 TI - Dosage-dependent requirement for mouse Vezf1 in vascular system development. AB - Vezf1 is an early development gene that encodes a zinc finger transcription factor. In the developing embryo, Vezf1 is expressed in the yolk sac mesoderm and the endothelium of the developing vasculature and, in addition, in mesodermal and neuronal tissues. Targeted inactivation of Vezf1 in mice reveals that it acts in a closely regulated, dose-dependent fashion on the development of the blood vascular and lymphatic system. Homozygous mutant embryos display vascular remodeling defects and loss of vascular integrity leading to localized hemorrhaging. Ultrastructural analysis shows defective endothelial cell adhesion and tight junction formation in the mutant vessels. Moreover, in heterozygous embryos, haploinsufficiency is observed that is characterized by lymphatic hypervascularization associated with hemorrhaging and edema in the jugular region; a phenotype reminiscent of the human congenital lymphatic malformation syndrome cystic hygroma. PMID- 15882862 TI - Combinatorial activity of Flamingo proteins directs convergence and extension within the early zebrafish embryo via the planar cell polarity pathway. AB - The seven-transmembrane protocadherin, Flamingo, functions in a number of processes during Drosophila development, including planar cell polarity (PCP). To assess the role(s) of Flamingo1/Celsr1 (Fmi1) during vertebrate embryogenesis we have exploited the zebrafish system, identifying two Fmi1 orthologues (zFmi1a and zFmi1b) and employing morpholinos to induce mis-splicing of zebrafish fmi1 mRNAs, to both imitate mutations identified in Drosophila flamingo and generate novel aberrant Flamingo proteins. We demonstrate that in the zebrafish gastrula, Fmi1 proteins function in concert with each other and with the vertebrate PCP proteins, Wnt11 and Strabismus, to mediate convergence and extension during gastrulation, without altering early dorso-ventral patterning. We show that zebrafish Fmi1a promotes extension of the entire antero-posterior axis of the zebrafish gastrula including prechordal plate and ventral diencephalic precursors. However, while we show that control over axial extension is autonomous, we find that Fmi1a is not required within lateral cells undergoing dorsal convergence. PMID- 15882863 TI - Membrane type 1-matrix metalloproteinase induces endothelial cell morphogenic differentiation by a caspase-dependent mechanism. AB - Membrane-type 1 matrix metalloproteinase (MT1-MMP) has been suggested to play an essential role in angiogenesis. Based on recent evidence suggesting that the sprouting and branching of capillaries during angiogenesis involves apoptosis, we investigated the involvement of this process in MT1-MMP-dependent morphogenic differentiation of EC into capillary-like structures. We found that MT1-MMP sensitizes EC to apoptosis, since reduction of MT1-MMP expression abolished vimentin fragmentation in apoptotic HUVECs while overexpression of the enzyme induced caspase-3 activity in BAECs subjected to pro-apoptotic treatments. MT1 MMP-mediated caspase-3 activation likely occurs through the mitochondrial pathway since it was abrogated by Bcl-2, but not by CrmA overexpression. Reduction of MT1 MMP expression in HUVECs reduced morphogenic differentiation that was correlated with diminished vimentin fragmentation, whereas its overexpression in BAECs stimulated both processes. Inactivation of the catalytic activity or removal of the cytoplasmic domain of MT1-MMP markedly reduced its ability to induce both morphogenic differentiation and caspase-3 activation. The inhibitory effects of the anti-apoptotic protein Bcl-2 and the caspase inhibitor zVAD-fmk further suggested the involvement of apoptosis during MT1-MMP-mediated morphogenic differentiation. Our results show that the ability of MT1-MMP to induce EC morphogenic differentiation involves its activation of a caspase-dependent mechanism. PMID- 15882864 TI - Skeletal muscle stem and progenitor cells: reconciling genetics and lineage. AB - Skeletal muscle provides a unique paradigm for studying stem to differentiated cell transitions, as well as the acquisition of cellular identity. Embryological and genetic studies over the last decades have unveiled key signaling pathways and regulatory genes which are involved in this process. In the adult, regeneration from fiber-associated satellite cells as well as non-muscle cells have opened the perspective for cell therapy studies. Paradoxically, however, the lineage has remained largely elusive. Recent studies have provided clues regarding the cellular organization in this lineage. Furthermore, the complexity of the genetic networks regulating global and local myogenic programs can be correlated with location and lineage. Finally, prenatal and postnatal developmental strategies have similarities and differences which will also be highlighted. PMID- 15882865 TI - Plasmodium falciparum: characterization of a late asexual stage golgi protein containing both ankyrin and DHHC domains. AB - Proteins containing the DHHC motif have been shown to function as palmitoyl transferases. The palmitoylation of proteins has been shown to play an important role in the trafficking of proteins to the proper subcellular location. Herein, we describe a protein containing both ankyrin domains and a DHHC domain that is present in the Golgi of late schizonts of P. falciparum. The timing of expression as well as the location of this protein suggests that it may play an important role in the sorting of proteins to the apical organelles during the development of the asexual stage of the parasite. PMID- 15882866 TI - Taenia saginata: differential diagnosis of human taeniasis by polymerase chain reaction-restriction fragment length polymorphism assay. AB - Speciation of Taenia in human stool is important because of their different clinical and epidemiological features. DNA analysis has recently become possible which overcomes the problems of differentiating human taeniid cestodes morphologically. In the present study, we evaluated PCR coupled to restriction fragment length polymorphism to differentiate Taenia solium from Taenia saginata eggs present in fecal samples from naturally infected patients. A different DraI RFLP pattern: a two-band pattern (421 and 100 bp) for T. saginata and a three band pattern (234, 188, and 99 bp) for T. solium was observed allowing the two species to be separated. The lower detection limit of the PCR-RFLP using a non infected fecal sample prepared with a given number of T. saginata eggs was 34 eggs in 2 g stool sediment. The 521 bp mtDNA fragment was detected in 8 out of 12 Taenia sp. carriers (66.6%). Of these, three showed a T. solium pattern and five a T. saginata pattern. PMID- 15882867 TI - Real-time PCR method for the quantitative analysis of human T-cell receptor gamma and beta gene rearrangements. AB - Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the alphabeta vs. gammadelta lineage decision, and characterizing T-cell leukemias. Methods such as PCR and quantitative Southern blotting provide useful information, but also have significant shortcomings such as lack of quantitation in the case of PCR and technical challenges in the case of Southern blotting. Here we describe a real-time PCR method that overcomes many of these shortcomings. This new method shows comparable results for the fraction of unrearranged TCRgamma and TCRbeta genes in human thymocytes and peripheral blood T cells as Southern blotting, and has the advantages of being simple to perform, highly quantitative, and requiring nanogram quantities of DNA. We also describe a real-time PCR method to quantitate T-cell receptor excision circles formed during TCRbeta rearrangements. PMID- 15882868 TI - Effect of Diplocystis tipulae Sherlock (Eugregarinida: Apicomplexa), a coelomic gregarine pathogen of tipulids, on the larval size of Tipula paludosa Meigen (Tipulidae: Diptera). AB - This study demonstrates the debilitative effect of a coelomic gregarine, Diplocystis tipulae, on Tipula paludosa. The larvae were provided contaminated fresh grass leaves from a field where 40.0% of T. paludosa larvae were infected by this pathogen. Resultant infected larvae were separated into four groups according to infection level. Analysing their weights, lengths, and weight/length ratios showed that larval size decreased as infection level increased. Differences, especially at the lower and upper levels of the infection levels, were statistically significant. It was concluded that infection by D. tipulae affected the size of T. paludosa larvae resulting in smaller individuals. PMID- 15882869 TI - Active site identification through geometry-based and sequence profile-based calculations: burial of catalytic clefts. AB - Electrostatics calculations with proteins that are uniformly charged over volume can aid enzyme/non-enzyme discrimination. For known enzymes, such methods locate active sites to within 5% on the enzyme surface, in 77% of a test set. We now report that removing the dielectric boundary improves active site location to 80%, with optimal discrimination between enzymes and non-enzymes of around 80% specificity and 80% sensitivity. This calculation quantifies burial of solvent accessible regions. Many of the true enzymes incorrectly assigned as non-enzymes have active sites at subunit boundaries. These are missed in monomer-based calculations. Catalytic and non-catalytic antibodies are studied in this context of active/binding site burial. Whilst catalytic antibodies, on average, have marginally higher active site burial than non-catalytic antibodies, these values are generally smaller than for non-antibody enzymes, possibly contributing to their relatively low turnover. Prediction of active site location improves further when sequence profile-based weights replace the uniform charge distribution, so that a combination of burial and amino acid conservation is assessed. Accuracy rises to 93% of active sites to within 5%, in the test set, for the optimal profile weights scheme. The equivalent value in a separate validation set is 89% to within 5%. Enzyme/non-enzyme and enzyme functional site predictions are made for structural genomics proteins, suggesting that a substantial majority of these are non-enzymes. PMID- 15882870 TI - Sequence dependence of amyloid fibril formation: insights from molecular dynamics simulations. AB - The clarification of the physico-chemical determinants underlying amyloid deposition is critical for our understanding of misfolding diseases. With this purpose we have performed a systematic all-atom molecular dynamics (MD) study of a series of single point mutants of the de novo designed amyloidogenic peptide STVIIE. Sixteen different 50ns long simulations using explicit solvent have been carried out starting from four different conformations of a polymeric six stranded beta-sheet. The simulations have provided evidence for the influence of a small number of site-specific hydrophobic interactions on the packing and stabilization of nascent aggregates, as well as the interplay between side-chain interactions and the net charge of the molecule on the strand arrangement of polymeric beta-sheets. This MD analysis has also shed light into the origin of the position dependence on mutation of beta-sheet polymerization that was found experimentally for this model system. Our results suggest that MD can be applied to detect critical positions for beta-sheet aggregation within a given amyloidogenic stretch. Studies similar to the one presented here can guide site directed mutations or the design of drugs that specifically disrupt the key stabilizing interactions of beta-sheet aggregates. PMID- 15882871 TI - Abnormally persistent latent inhibition induced by lesions to the nucleus accumbens core, basolateral amygdala and orbitofrontal cortex is reversed by clozapine but not by haloperidol. AB - Latent inhibition (LI) is the proactive interference of inconsequential preexposure to a stimulus with its ability to signal significant events, and disrupted LI is considered to model positive symptoms of schizophrenia. We have recently shown that lesions of the nucleus accumbens core (NACc), basolateral amygdala (BLA) and orbitofrontal cortex (OFC) produce abnormally persistent LI, and suggested that this phenomenon may model negative symptoms. Here we tested whether NACc, BLA and OFC lesion-induced persistent LI would be reversed by the atypical antipsychotic drug (APD) clozapine but not by the typical APD haloperidol. Because clozapine's action is likely reflecting its 5HT2A receptor antagonism, we also tested whether NACc lesion-induced persistent LI would be reversed by the selective 5HT2A antagonist M100907. LI was measured in a conditioned emotional response procedure by comparing suppression of drinking in response to a tone in rats receiving 0 (non-preexposed) or 40 tone presentations (preexposed) followed by five tone-shock pairings. Under these conditions, control rats did not show LI but all lesioned rats persisted in exhibiting LI, and this was reversed by clozapine but not by haloperidol. In addition, M100907 reversed NACc lesion-induced persistent LI. These two novel phenomena, abnormally persistent LI and its selective reversal by an atypical APD, suggest a novel index of schizophrenia relevant behavioral abnormality and of atypical antipsychotic activity in the LI model. The identification of brain regions whose damage leads to persistent LI in the rat may provide valuable cues on dysfunctional brain circuits involved in negative symptoms and in the action of atypical APDs. PMID- 15882872 TI - Error processing in major depressive disorder: evidence from event-related potentials. AB - In a previous study we showed that errors following errors activate a strategic (prefrontal) mechanism. In an error trial (trial n) following an erroneous previous trial (trial n - 1) healthy control subjects were found to have enlarged (more negative) amplitudes of the error related negativity (ERN)/error negativity (Ne), an electrophysiological correlate of response monitoring, in response to a negative feedback signal. Contrary to that, patients with major depressive disorder showed smaller (less negative) ERN/Ne amplitudes. It has been discussed controversially whether errors of choice (e.g., pressing an incorrect response button in an Eriksen flanker task) and errors of commission (e.g., pressing a button when one is not supposed to in a Go/Nogo task) are related to different ERN/Ne mechanisms. In the present study, we examined whether our previous result only holds for errors of choice in an Eriksen flankers task or extends to errors of commission in a Go/Nogo task, as well. Ten patients with DSM-IV major depressive disorder and 10 matched controls participated in a Go/Nogo task with performance feedback which signaled monetary reward. Patients with major depressive disorder again showed a less negative ERN/Ne amplitude in error trials following error trials. This result might reflect impaired response monitoring processes in major depressive disorder resulting from an impaired activation of a central reward pathway and/or a deficit in strategic reasoning. PMID- 15882873 TI - Household youth on minority operated farms in the United States, 2000: exposures to and injuries from work, horses, ATVs and tractors. AB - INTRODUCTION: It is likely that youth living on minority operated farms (<3% of U.S. farms) face hazards similar to the general farm population; however, since minority youth are not well represented by general farm surveys, this information hasn't been confirmed. METHOD: Nonfatal injury and exposure data were obtained from the 2000 Minority Farm Operator Childhood Agricultural Injury Survey (M CAIS). RESULTS: On racial minority farms, there were an estimated 28,600 household youth. Of these, about 41% worked, 26% rode a horse, 23% drove an ATV, and 23% operated a tractor. On Hispanic farms, there were an estimated 17,998 household youth. Of these, 44% worked, 30% rode a horse, 27% drove an ATV, and 25% operated a tractor. CONCLUSIONS: These results show the value of conducting a survey of minorities to identify high risk groups and target issues that may be unique to the minority farm population. PMID- 15882874 TI - Is road rage increasing? Results of a repeated survey. AB - PROBLEM: We report on trends in road rage victimization and perpetration based on population survey data. METHOD: Based on repeated cross-sectional telephone surveys of Ontario adults between July 2001 and December 2003, logistic regression analyses examined differences between years in road rage victimization and perpetration in the previous year controlling for demographic characteristics. RESULTS: The prevalence of any road rage victimization in the previous year decreased significantly from 47.5% in 2001 to 40.6% in 2003, while prevalence of any road rage perpetration remained stable (31.0% to 33.6%). Logistic regression analyses revealed that the odds of experiencing any road rage victimization was 33% higher in 2001 and 30% higher in 2002, than in 2003. DISCUSSION: Survey data provide a valuable perspective on road rage trends, but efforts to track road rage incidents is also needed. SUMMARY: In Ontario, the proportion of adults experiencing any road rage victimization decreased from 2001 to 2003 while the proportion reporting any road rage perpetration remained stable. IMPACT ON INDUSTRY: None. PMID- 15882875 TI - Incidence rates and risk factors for injury events in high school students in Lijin County, China. AB - PROBLEM: Most investigations of injuries among Chinese populations are focused on urban or better developed areas, fewer on rural or poor areas. METHODS: From September 21 to 27, 2002, an epidemiological investigation of eight kinds of injury-related events was carried out among 1,944 students in 39 classes randomly sampled from middle schools in Lijin County, a rural area. The eight kinds of events were: (a) traffic accidents, (b) poisoning, (c) fall, (d) burn or scald, (e) blunt or sharp tools contact, (f) electrical contact, (g) animal or insect bites, and (h) non-fatal drowning. RESULTS: The total incidence rate of eight kinds of injuries in the previous year was 21.1%. The incidence rate among boys was 24.2%, while that among girls was 17.4%, which was significant (chi square=13.4, P<0.05). The incidence rate among junior high school students was 23.1%, while that among senior high school students was 15.5%, which was also significant (chi-square=13.2, P<0.05). A case-control study of 410 students with at least one of eight kinds of injury events indicated that students' gender and their mothers' health status were related to their injuries. CONCLUSION: The injury incidence rate in a rural area was less than other results in urban or developed areas. Students' gender and mothers' health status were important factors associated with the occurrence of injury. IMPACT ON INDUSTRY: In order to improve the surroundings of the growing population of adolescents, some preventive measures should be put forward, especially to the high-risk students of injury. PMID- 15882876 TI - Relocation of truncated bid plays an important role in suppression of tumor necrosis factor alpha induced apoptosis in hepatocytes isolated from transgenic mouse. AB - BACKGROUND/AIMS: Tumor necrosis factor alpha (TNFalpha) induces apoptosis in murine hepatocytes pretreated with Actinomycin D (ActD) in vitro. This study sought to clarify the relationship between hepatic energy status and TNFalpha induced hepatocyte apoptosis, using mice transgenic for creatine kinase (CK) expression. MATERIALS AND METHODS: Hepatocytes from CK transgenic mice were cultured with or without creatine (Cr). The concentrations of ATP and phosphocreatine (PCr) in hepatocytes were measured by high-pressure liquid chromatography. Sixteen hours after treatment with ActD and TNFalpha, we evaluated cell viability of these hepatocytes. We examined truncated Bid and cytochrome c by immunoblot analysis. RESULTS: Six hours after cell isolation, the concentration of PCr in CK transgenic hepatocytes cultured with Cr increased to 8.23 +/- 0.01 microg/mg protein, while that of hepatocytes cultured without Cr was lower than 0.1 microg/mg protein. In hepatocytes cultured without Cr, ActD and TNFalpha treatment induced massive cell death, while hepatocytes cultured with Cr maintained greater than 80% viability. In CK transgenic hepatocytes cultured with Cr, truncated Bid relocation to mitochondria was highly suppressed, compared to CK transgenic hepatocytes cultured without Cr. CONCLUSION: PCr accumulation may prevent TNFalpha-induced apoptosis in murine hepatocytes by suppression of truncated Bid targeting to mitochondria. PMID- 15882877 TI - Effects of dehydroepiandrosterone administration on rat hepatic metabolism following thermal injury. AB - BACKGROUND: Severe burns cause dramatic alterations in liver and whole-body metabolism. Recently, there has been interest in using dehydroepiandrosterone (DHEA) as a treatment for trauma patients, and enhanced survival and immune function have been reported using DHEA in animal trauma models. The specific effects of DHEA on hepatic metabolism following burn injury have not been explored. MATERIALS AND METHODS: Male rats received either (1) a burn covering approximately 20% of the total body surface area or a sham burn or (2) burn injury followed by two intraperitoneal injections of DHEA or vehicle. After 4 days, the livers were isolated and perfused in vitro, and 28 metabolite fluxes were measured. Metabolic flux analysis was used to obtain the intracellular metabolic flux distribution and provide an overview of the metabolic state of the livers in each experimental group. RESULTS: Burn injury decreased the uptake of lactate and the production of beta-hydroxybutyrate and increased the deamination of glutamine to glutamate and asparagine to aspartate. DHEA, compared to vehicle treatment, decreased pentose phosphate pathway (PPP) fluxes and the uptake of several amino acids in burned rats. Furthermore, DHEA treatment restored liver metabolism in burned rats to a state that was very similar to that of the sham control group. CONCLUSIONS: DHEA administration appears to normalize hepatocellular metabolism in burned rats but also decreases the PPP flux, which may impair the liver's ability to recycle endogenous antioxidants. DHEA treatment combined with exogenous antioxidants should receive further consideration in the management of burn and trauma patients. PMID- 15882878 TI - VEGF-mediated survivin expression in neuroblastoma cells. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) up-regulates a number of cellular survival signals in endothelial cells. We hypothesize that VEGF will up regulate survivin, a member of the IAP family of anti-apoptotic proteins, via the PI3K/Akt cell signaling pathway in human neuroblastoma cells. MATERIALS AND METHODS: IMR-32 human neuroblastoma cells are cultured with VEGF at varying times and in escalating doses. A specific inhibitor of PI3-kinase, LY294002, is used to block Akt phosphorylation. Immunoblot is used to measure protein expression, and Hoechst staining is used to detect apoptosis. RESULTS: Stimulation of IMR-32 neuroblastoma cells with VEGF results in an increase in survivin protein expression in both a dose- and a time-dependent fashion. Akt phosphorylation is also increased after stimulation with exogenous VEGF. Blockade of Akt phosphorylation with LY294002 abrogates the effects of VEGF upon survivin and phosphorylated Akt protein expression. CONCLUSIONS: VEGF has been shown to up regulate a number of survival signals in endothelial cells. We have found that exposure of human neuroblastoma cells to exogenous VEGF results in an increased expression of survivin protein and phosphorylated Akt, and inhibition of PI3 kinase abrogates those effects. It appears that VEGF is important for promotion of neuroblastoma cellular survival through the up-regulation of survival proteins, and not only through its angiogenic properties. PMID- 15882879 TI - Effect of pre-operative short course famotidine on tumor infiltrating lymphocytes in colorectal cancer: a double blind, placebo controlled, prospective randomized study. AB - BACKGROUND: Pilot studies have shown that histamine H2 receptor antagonists augment the natural immunity against cancer in colorectal and gastric cancer by enhancing lymphocytic infiltration in the tumors. However, a study of adjuvant ranitidine failed to show a significant benefit in colorectal cancer, possibly because of the immunosuppression exerted by blood transfusion and post-operative infections. The pre-operative use of H2 receptor antagonists may therefore be of greater benefit. Except for a pilot study using cimetidine, there are no trials that have evaluated the effect of pre-operative H2 receptor antagonists on tumor infiltrating lymphocytes in colorectal cancer. OBJECTIVE: To evaluate the efficacy of famotidine in augmenting tumor infiltrating lymphocytes in colorectal cancer. STUDY DESIGN: Double blind, placebo controlled, prospective randomized study. METHODS: Twenty-three patients with resectable colorectal cancer were randomized to receive famotidine (n = 11) or placebo (n = 12). Famotidine was given for 1 week pre-operatively in a dose of 40 mg per day p.o. After resection, the specimens were analyzed histologically for lymphocytic infiltration by a pathologist blinded to the two groups. Lymphocytic infiltration more than 50 cells per high power field, involving more than 50% of the tumor-normal tissue interface was considered significant. RESULTS: The two groups were comparable for age, gender, pre-operative carcino embryonic antigen (CEA) levels and pathological stage. Significant lymphocytic infiltration was seen in 63.6% (7 of 11) patients in the study group compared to only 8.5% (1 of 12) patients in the placebo group (P = 0.005). Despite fewer recurrences and a longer survival in the study group, the difference was not significant. CONCLUSION: This study shows that pre-operative famotidine may significantly enhance lymphocytic infiltration in colorectal cancer and may have potential for use as an anticancer agent in colorectal cancer. PMID- 15882880 TI - Steroids and brain atrophy in multiple sclerosis. AB - In this review, we focus on different pathogenetic mechanisms of corticosteroids that induce short- and long-term brain volume fluctuations in a variety of systemic conditions and disorders, as well as on corticosteroid-induced immunomodulatory, immunosuppressive and anti-inflammatory mechanisms that contribute to the slowdown of brain atrophy progression in patients with multiple sclerosis (MS). It appears that chronic low-dose treatment with corticosteroids may contribute to irreversible loss of brain tissue in a variety of autoimmune diseases. This side effect of steroid therapy is probably mediated by steroid induced protein catabolism mechanism. Evidence is mounting that high-dose corticosteroids may induce reversible short-term brain volume changes due to loss of intracellular water and reduction of abnormal vascular permeability, without there having been axonal loss. Other apoptotic and selective inhibiting mechanisms have been proposed to explain the nature of corticosteroid-induced brain volume fluctuations. It has been shown that chronic use of high dose intravenous methylprednisolone (IVMP) in patients with MS may limit brain atrophy progression over the long-term via different immunological mechanisms, including downregulation of adhesion molecule expression on endothelial cells, decreased cytokine and matrix metalloproteinase secretion, decreased autoreactive T-cell mediated inflammation and T-cell apoptosis induction, blood-brain barrier closure, demyelination inhibition and, possibly, remyelination promotion. Studies in nonhuman primates have confirmed that short-term brain volume fluctuations may be induced by corticosteroid treatment, but that they are inconsistent, potentially reversible and probably dependent upon individual susceptibility to the effects of corticosteroids. Further longitudinal studies are needed to elucidate pathogenetic mechanisms contributing to brain volume fluctuations in autoimmune diseases and multiple sclerosis. PMID- 15882881 TI - Repeated stress effects on nociception and on ectonucleotidase activities in spinal cord synaptosomes of female rats. AB - It has been reported that animals submitted to repeated restraint stress present various adaptation responses which are dependent on the sex. These adaptations include changes in nociception and adenine nucleotide hydrolysis. In this study, we report the effect of chronic administration of a gonadal steroid (17beta estradiol) on ATP, ADP and AMP hydrolysis in spinal cord synaptosomes of adult ovariectomized (OVX) Wistar rats submitted to repeated restraint stress over 40 days. We also measured nociceptive threshold in these animals using the tail flick test. The results show that tail-flick latencies were decreased in both stressed groups, OVX and OVX rats receiving estradiol replacement therapy, indicating reduced nociceptive threshold after exposure to repeated stress. Repeated restraint stress caused no effect on ATPase or ADPase activities. On the other hand, AMP hydrolysis in spinal cord synaptosomes from repeatedly stressed rats was decreased in OVX rats compared to non-stressed OVX ones, indicating reduced extracellular adenosine production; this effect was reversed by hormonal replacement. These observations suggest that nociceptive sensitivity to noxious stimuli is affected by repeated stress and that modulation of neurotransmission by adenine nucleotides in spinal cord may be altered by the interaction of sexual hormones and psychological factors, such as exposure to stress. PMID- 15882882 TI - Capsaicin stimulates the non-store-operated Ca2+ entry but inhibits the store operated Ca2+ entry in neutrophils. AB - Rat neutrophils express the mRNA encoding for transient receptor potential (TRP) V1. However, capsaicin-stimulated [Ca2+]i elevation occurred only at high concentrations (> or = 100 microM). This response was substantially decreased in a Ca2+-free medium. Vanilloids displayed similar patterns of Ca2+ response with the rank order of potency as follows: scutigeral>resiniferatoxin>capsazepine>capsaicin=olvanil>isovelleral. Arachidonyl dopamine (AAD), an endogenous ligand for TRPV1, failed to desensitize the subsequent capsaicin challenge. Capsaicin-induced Ca2+ response was not affected by 8-bromo-cyclic ADP-ribose (8-Br-cADPR), the ryanodine receptor blocker, but was slightly attenuated by 1-[6-[17beta-3-methoxyestra-1,3,5(10)-trien-17 yl]amino]hexyl]-1H-pyrrole-2,5-dione (U-73122), the inhibitor of phospholipase C coupled processes, 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H imidazole (SKF-96365), the blocker of receptor-gated and store-operated Ca2+ (SOC) channels, 2-aminoethyldiphenyl borate (2-APB), the blocker of D-myo inositol 1,4,5-trisphospahte (IP3) receptor and Ca2+ influx, and by ruthenium red, a blocker of TRPV channels, and enhanced by the Ca2+ channels blocker, cis-N (2-phenylcyclopentyl)azacyclotridec-1-en-2-amine (MDL-12330A) and Na+ deprivation. In addition, capsaicin had no effect on the plasma membrane Ca2+ ATPase activity or the production of nitric oxide (NO) and reactive oxygen intermediates (ROI) or on the total thiols content. Capsaicin (> or = 100 microM) inhibited the cyclopiazonic acid (CPA)-induced store-operated Ca2+ entry (SOCE). In the absence of external Ca2+, the robust Ca2+ entry after subsequent addition of Ca2+ was decreased by capsaicin in CPA-activated cells. Capsaicin alone increased the actin cytoskeleton, and also increased the actin filament content in cell activation with CPA. These results indicate that capsaicin activates a TRPV1-independent non-SOCE pathway in neutrophils. The reorganization of the actin cytoskeleton is probably involved in the capsaicin inhibition of SOCE. PMID- 15882883 TI - Oxidative stress, mitochondrial permeability transition, and cell death in Cu exposed trout hepatocytes. AB - We have previously shown that, in trout hepatocytes, exposure to a high dose of copper (Cu) leads to disruption of Ca(2+) homeostasis and elevated formation of reactive oxygen species (ROS), with the latter ultimately causing cell death. In the present study, we aimed at identifying, using a lower Cu concentration, the role of mitochondria in this scenario, the potential involvement of the mitochondrial permeability transition (MPT), and the mode of cell death induced by the metal. Incubation with 10 muM Cu resulted in a strong stimulation of ROS formation, and after 2 h of exposure a significant increase of both apoptotic and necrotic cells was seen. Co-incubation of Cu-treated hepatocytes with the iron chelator deferoxamine significantly inhibited ROS production and completely prevented cell death. The origin of the radicals generated was at least partly mitochondrial, as visualized by confocal laser scanning microscopy. Furthermore, ROS production was diminished by inhibition of mitochondrial respiration, but since this also aggravated the elevation of intracellular Ca(2+) induced by Cu, it did not preserve cell viability. In a sub-population of cells, Cu induced a decrease of mitochondrial membrane potential and occurrence of the MPT. Cyclosporin A, which did not inhibit ROS formation, prevented the onset of the MPT and inhibited apoptotic, but not necrotic, cell death. Cu-induced apoptosis therefore appears to be dependent on induction of the MPT, but the prominent contribution of mitochondria to ROS generation also suggests an important role of mitochondria in necrotic cell death. PMID- 15882884 TI - Snake venom fibrin(ogen)olytic enzymes. AB - Snake venoms contain a number of serine and metalloproteinases and included among these are the fibrin(ogen)olytic proteinases. Some years ago it was postulated that the fibrin(ogen)olytic enzymes may be clinically useful. Over the past 150 years a substantial body of literature has been generated on the identification and characterization of fibrin(ogen)olytic enzymes from a broad spectrum of snake species. In this review we describe the two different classes of fibrin(ogen)olytic enzymes isolated from snake venom and we summarize a number of studies aimed at characterizing the purified enzymes and/or their derivatives. Two distinct classes of venom fibrin(ogen)olytic enzymes have been previously identified, the metalloproteinases and serine proteinases. These two classes of proteinases differ in their mechanism of action and they target different amino acid sequences in fibrin(ogen), but each perform the same role in nature. When a snake envenomates its prey it needs a mechanism to facilitate the spread of the toxic components throughout the circulation. Fibrin(ogen)olytic enzymes break down fibrin rich clots and help to prevent further clot formation by their action on fibrinogen. This characteristic feature has led to development of fibrin(ogen)olytic snake venom enzymes as potential clinical agents to treat occlusive thrombi. Fibrolase, a fibrinolytic metalloproteinase isolated from Agkistrodon contortrix contortrix venom and the serine beta-fibrinogenolytic proteinase from Vipera lebetina have been chosen as representative enzymes from the two classes, and their biochemical and physiochemical properties will be described in detail. Finally, the characterization and development of alfimeprase, a recombinant fibrinolytic enzyme derived from fibrolase, as a clinical agent is described citing the progression from the laboratory bench to its current status as having successfully completed Phase II clinical trials. PMID- 15882885 TI - Dominant negative selection of vaccinia virus using a thymidine kinase/thymidylate kinase fusion gene and the prodrug azidothymidine. AB - The Escherichia coli thymidine kinase/thymidylate kinase (tk/tmk) fusion gene encodes an enzyme that efficiently converts the prodrug 3'-azido-2',3' dideoxythymidine (AZT) into its toxic triphosphate derivative, a substance which stops DNA chain elongation. Integration of this marker gene into vaccinia virus that normally is not inhibited by AZT allowed the establishment of a powerful selection procedure for recombinant viruses. In contrast to the conventional vaccinia thymidine kinase (tk) selection that is performed in tk-negative cell lines, AZT selection can be performed in normal (tk-positive) cell lines. The technique is especially useful for the generation of replication-deficient vaccinia viruses and may also be used for gene knock-out studies of essential vaccinia genes. PMID- 15882886 TI - Uncoating of HIV-1 requires cellular activation. AB - Uncoating is an essential step in viral replication cycle. Little is known about the mechanism and requirement of HIV uncoating. Using an in vitro uncoating model, we demonstrate here that the uncoating of HIV-1 was efficiently induced by lysate from activated CD4+ lymphocytes, while quiescent CD4+ lymphocyte lysate was unable to uncoat HIV-1 core. The uncoating activity was associated with an induction of in vitro reverse transcription of the viral genome. Using CD4+ lymphocytes that were arrested in cell cycle, we showed that the uncoating activity required transition of cells from G(0)/G(1a) into G(1b) stage. These results strongly suggested a requirement of cell cycle-dependent specific factors for HIV-1 uncoating. The putative HIV-1 uncoating factors could be fractionated from cell lysate by gel filtration chromatography. PMID- 15882887 TI - Homeostasis and function of goblet cells during rotavirus infection in mice. AB - Rotaviruses are the leading cause of severe viral gastroenteritis in young children. To gain insight in goblet cell homeostasis and intestinal mucin expression during rotavirus infection, 6-day-old mice were inoculated with murine rotavirus. To determine epithelial cell migration, mice were injected with BrdU just before inoculation. Small intestines were isolated at different days postinfection (dpi) and evaluated for rotavirus and goblet cell-specific gene expression. Small intestinal mucins of control and infected animals at 1, 2, and 4 dpi were isolated and tested for their capability to neutralize rotavirus infection in vitro. After inoculation, two peaks of viral replication were observed at 1 and 4 dpi. During infection, the number of goblet cells in infected mice was decreased in duodenum and jejunum, but was unaffected in the ileum. Goblet cells in infected animals accumulated at the tips of the villi. Muc2 mRNA levels were increased during the peak of viral replication at 1 dpi, whereas at other time points Muc2 and Tff3 mRNA levels were maintained at control levels. Muc2 protein levels in the tissue were also maintained, however Tff3 protein levels were strongly decreased. The number of goblet cells containing sulfated mucins was reduced during the two peaks of infection. Mucins isolated at 1 and 2 dpi from control and infected mice efficiently neutralized rotavirus infection in vitro. Moreover, mucins isolated from infected mice at 4 dpi were more potent in inhibiting rotavirus infection than mucins from control mice at 4 dpi. In conclusion, these data show that during rotavirus infection, goblet cells, in contrast to enterocytes, are relatively spared from apoptosis especially in the ileum. Goblet cell-specific Muc2 expression is increased and mucin structure is modified in the course of infection. This suggests that goblet cells and mucins play a role in the active defense against rotavirus infection and that age dependent differences in mucin quantities, composition, and/or structure alter the anti-viral capabilities of small intestinal mucins. PMID- 15882888 TI - A semi-implicit finite element model for natural water bodies. AB - In this work, a semi-implicit model, applied to the shallow water equations, is developed for natural water bodies. In the proposed model, the shallow water equations are integrated in the vertical direction, finite elements are employed in the spatial discretization, and finite differences in the time discretization. The model is based on the uncoupling of the governing equations, in which the expressions of the velocity components, obtained explicitly from the discretized momentum equations, are substituted in the continuity equation. Therefore, there is an uncoupling of the solution; initially the continuity equation is solved, and, in the sequence, the momentum equations are solved. Yet, this uncoupling produces a significant reduction in the number of equations of the resulting systems, when compared to standard coupled systems. This reduction improves substantially the computer performance. PMID- 15882889 TI - Removal of disinfection by-product precursors with ozone-UV advanced oxidation process. AB - The efficacy of using ozone (O3), ultraviolet irradiation (UV) and the combined O3-UV advanced oxidation process (AOP) to remove 2 classes of disinfection by product (DBP) precursors from raw surface water samples have been evaluated and compared. In particular, trihalomethane and haloacetic acids formation potentials were measured. Laboratory batch scale experiments were carried out as a function of ozone and UV dosage in order to study the removal kinetics. It is concluded that the combined O3-UV AOP is more effective than either the ozone or UV treatment alone. Ozone-UV AOP is capable of mineralizing up to 50% of the total organic carbon from the raw source water at an ozone dose of 0.62+/-0.019 mg O3/mL and a UV dose of 1.61 W s/cm2. In addition, O3-UV AOP can reduce trihalomethane formation potential by roughly 80% and haloacetic acids formation potential by roughly 70% at the same ozone and UV dosage. PMID- 15882890 TI - Reaction pathways and mechanisms of the electrochemical degradation of phenol on different electrodes. AB - Laboratory experiments were carried out on the kinetics and pathways of the electrochemical (EC) degradation of phenol at three different types of anodes, Ti/SnO2-Sb, Ti/RuO2, and Pt. Although phenol was oxidised by all of the anodes at a current density of 20 mA/cm2 or a cell voltage of 4.6 V, there was a considerable difference between the three anode types in the effectiveness and performance of EC organic degradation. Phenol was readily mineralized at the Ti/SnO2-Sb anode, but its degradation was much slower at the Ti/RuO2 and Pt anodes. The analytical results of high-performance liquid chromatography (HPLC) and gas chromatography coupled with mass spectrometry (GC/MS) indicated that the intermediate products of EC phenol degradation, including benzoquinone and organic acids, were subsequently oxidised rapidly by the Ti/SnO2-Sb anode, but accumulated in the cells of Ti/RuO2 and Pt. There was also a formation of dark coloured polymeric compounds and precipitates in the solutions electrolyzed by the Ti/RuO2 and Pt anodes, which was not observed for the Ti/SnO2-Sb cells. It is argued that anodic property not only affects the reaction kinetics of various steps of EC organic oxidation, but also alters the pathway of phenol electrolysis. Favourable surface treatment, such as the SnO2-Sb coating, provides the anode with an apparent catalytic function for rapid organic oxidation that is probably brought about by hydroxyl radicals generated from anodic water electrolysis. PMID- 15882891 TI - Endocrine response of the freshwater teleost, Sarotherodon mossambicus (Peters) to dimecron exposure. AB - The endocrine response in a freshwater teleost, Sarotherodon mossambicus (Peters) under dimecron (an organophosphate pesticide) toxicity was investigated by estimating the serum levels of T3 (triiodothyronine), T4 (thyroxine), cortisol, prolactin and insulin in control and sub-lethal (0.001 ml l(-1)) dimecron-exposed fish for 1, 6, 12, 24h and 5 days. In control S. mossambicus, the serum levels of T3 ranged from 0.80+/-0.01 to 0.82+/-0.01 ng ml(-1); T4 from 2.20+/-0.01 to 2.25+/-0.01 microg dl(-1); cortisol from 8.30+/-0.03 to 8.34+/-0.01 microg dl( 1); prolactin from 1.50+/-0.01 to 1.54+/-0.01 microg ml(-1); insulin from 9.70+/ 0.01 to 9.76+/-0.01 microU ml(-1) up to a maximum period of 5 days maintained in pollutant-free tap water. Exposure of fish to sub-lethal concentration of dimecron caused varying changes in the levels of serum hormones studied. Based on the results obtained, it was concluded that (i) the fish adaptively maintains a probable low metabolic rate, as indicated by the reduced levels of thyroid hormone (T3) as well as the glucocorticoid hormone (cortisol), which could be considered advantageous for the fish to indirectly reduce the toxic impact of the pesticide, (ii) the elevated levels of prolactin in the fish under pesticide stress is indicative of a possible hydromineral regulatory effect of the hormone (probably by influencing specific organs such as gills and kidney) under pesticide toxicity, (iii) the increased insulin level in the fish under pesticide stress is indicative of its role in favouring an adaptive tissue glycogenesis besides a possible increased lipogenesis to sequester the pesticide residue thereby reducing the toxic effect of the pesticide and (iv) the prolonged exposure of the fish (for 5 days) to sub-lethal dimecron appeared to exhibit a uniform recovery response in the different hormonal levels of the fish. PMID- 15882892 TI - Trace element and Sigma DDT concentrations in horticultural soils from the Tasman, Waikato and Auckland regions of New Zealand. AB - The long-term routine use of agrichemicals can result in elevated levels of trace elements and persistent organic pollutants in soils. Trace element concentrations and SigmaDDT levels were measured in soil (0-7.5 cm) samples collected from horticultural and grazing properties in 3 regions of New Zealand (Auckland, Tasman and Waikato). Elevated levels of arsenic (<2 to 58 mg kg(-1)), cadmium (<0.1 to 1.5 mg kg(-1)), copper (5 to 523 mg kg(-1)), lead (5 to 243 mg kg(-1)) and SigmaDDT (<0.03 to 34.5 mg kg(-1)) were detected in soils from all 3 regions. With the exception of cadmium and zinc, significantly higher levels of contaminants were generally detected in horticultural soils than in grazing soils. Our results have implications for the on-going use of agrichemicals as concentrations of cadmium, copper, tin and zinc in some samples exceeded ecotoxicity based soil criteria. The p,p'-DDE:DDT ratios indicate that the degradation of DDT in NZ horticultural soils may be inhibited by the co contamination with trace elements. PMID- 15882893 TI - Venous thromboembolism among United States soldiers deployed to Southwest Asia. AB - INTRODUCTION: Military operations may represent a high-risk environment for venous thromboembolism (VTE). We sought to identify and describe cases of venous thromboembolism among US military personnel serving in Southwest Asia, and estimate relative disease rates compared to non-deployed personnel. MATERIALS AND METHODS: Retrospective review of imaging archives, hospital discharge codes, case logs and autopsy records for the diagnosis of deep vein thrombosis or pulmonary embolism occurring from 1 March 2003 through 29 February 2004 among U.S. military personnel deployed to Southwest Asia. Rates of disease in deployed and non deployed active-duty soldiers were estimated using personnel data and deployment experience obtained from automated rosters. RESULTS: Forty cases of venous thromboembolism were identified. The case-fatality rate was 16% (3/19) among those with pulmonary embolism. Antecedent trauma followed by prolonged air evacuation was present in 55% (22/40). Compared to trauma-associated cases, non trauma cases were more commonly over 40 years old (44% vs. 5%; p<0.05), assigned to a transportation or quartermaster company (56% vs. 14%; p<0.05), or had a history of remote venous thromboembolism (31% vs. 0%; p<0.05). The overall incidence among deployed active-duty soldiers was 22.1/100,000 person-years. Compared to non-deployed active-duty soldiers, the age-adjusted incidence rate ratio was 1.06 (CI(0.95) 0.68-1.67). CONCLUSIONS: VTE rates among deployed soldiers are relatively low compared to the general population, and are comparable to non-deployed soldiers. Fatalities from PE are not uncommon, and vigilance among clinicians remains warranted. Trauma followed by prolonged air evacuation or ground transport during military operations may represent unique interactive risk factors for venous thromboembolism. PMID- 15882894 TI - Effect of the novel direct factor Xa inhibitor DX-9065a on thrombin generation and inhibition among patients with stable atherosclerotic coronary artery disease. AB - INTRODUCTION: Thrombin, a pluripotential effector enzyme with prothrombotic, proinflammatory, and mitogenic properties, plays a pivotal role in the pathobiology and clinical expression of atherothrombotic coronary artery disease. Existing anticoagulant drugs have not been shown to attenuate thrombin generation or activity consistently. We sought to investigate the effect of DX-9065a on thrombin generation and inhibition in patients with stable CAD. DX-9065a is a small-molecule, synthetic, direct inhibitor of factor Xa. MATERIALS AND METHODS: Peripheral venous blood samples were collected serially during and after administration of either placebo or 1 of 4 weight-adjusted regimens of DX-9065a, in 73 patients with stable CAD participating in the XaNADU-1B study. RESULTS AND CONCLUSIONS: At baseline, the median (25th, 75th) prothrombin activation fragment 1.2 (F1.2) level was 2.56 (2.05, 3.20) nmol/L, and the median d-dimer level was 0.26 (0.19, 0.38) microg FEU/L. There were significant relationships between measured plasma DX-9065a concentrations and both F1.2 (4.9% decrease for each doubling of DX-9065a) (P<0.0001) and d-dimer (5.5% decrease for each doubling of DX-9065a) (P=0.001). F1.2 was suppressed (below baseline) at 96 h after administration of DX-9065a. Coronary thrombotic events did not occur during or after study drug administration. DX-9065a, the first in a class of small molecule, direct, selective and reversible factor Xa inhibitors, reduces thrombin generation and fibrin formation among patients with stable CAD. The effect is concentration-dependent and persists for at least 96 h following drug cessation, without biochemical or clinical evidence of rebound. PMID- 15882895 TI - Phase II trial of capecitabine in recurrent squamous cell carcinoma of the cervix. AB - PURPOSE: To determine the efficacy and safety of capecitabine in women with inoperable, recurrent, or metastatic squamous cell cervical cancer. PATIENTS AND METHODS: In a phase II IRB approved trial, capecitabine was given at a dosage of 2000 mg/m2/day orally in a divided dose daily for 14 days followed by a 7-day rest period. A standard dose modification scheme was used with one allowed dose reduction or dose escalation. National Cancer Institute criteria for progression, response, and toxicity were utilized. Quality of life data were obtained using the Memorial Symptom Assessment Scale and Functional Assessment for Cancer Therapy, which included a subscale for cervical cancer. RESULTS: Twenty of 23 enrolled patients were evaluable for response. Stable disease was noted in 5 patients, with a median duration of response of 3.5 months (range, 3-6.5 months). No partial or complete responses were seen. Common grade 3 toxicities were fatigue (30.4%); abdominal pain, constipation, hand-foot syndrome, nausea, and vomiting (8.7% each); as well as dyspnea, headache, and coagulopathy (4.3% each). There were no grade 4 toxicities. All patients with previous exposure to infused 5-FU had evidence of progression. No statistically significant changes in quality of life were noted from baseline to post-cycle 2. CONCLUSION: Single-agent capecitabine in patients with recurrent cervical cancer resulted in no objective responses. Although capecitabine is a well-tolerated regimen, as a single agent, it offers minimal benefit in a poor-prognosis cervical cancer population. PMID- 15882896 TI - Role of molecular isoforms of acetylcholinesterase in learning and memory functions. AB - In the present study, activity of salt soluble (SS) G1 and detergent soluble (DS) G4 molecular isoforms of acetylcholinesterase (AChE) has been investigated in rat brain areas in trained (learned), scopolamine (amnesic) and Tacrine (anti dementic) treated rats to find out their role in learning and memory functions. AChE was estimated spectrophotometrically at 412 nm in rat brain areas. Isolation and partial purification of molecular isoforms G1 and G4 of AChE was done by gel filtration chromatography. Passive avoidance was used to test learning and memory functions. AChE activity was altered in both the fractions SS and DS of different brain areas following passive avoidance in control, scopolamine treated, tacrine treated and tacrine treatment in scopolamine pretreated rats. The peak AChE activity obtained in the DS (fraction 9) and the SS (fraction 13) fraction following gel filtration chromatography. On the basis of molecular weight fraction 9 (DS) and 13 (SS) represent the G4 and G1, respectively. The pattern of changes in the AChE activity of G1 isoform (fraction 13 of SS) and G4 isoform (fraction 9 of DS) in brain areas were similar to those of SS and DS fraction, respectively. In hippocampus, AChE activity in the fraction G1 isoform (fraction 13 of SS) was decreased only in tacrine treated rats but AChE activity in the G4 isoform (fraction 9 of DS) was decreased in both trained and tacrine treated rats. Changes in activity of G4 isoform of AChE in hippocampus could be correlated with passive avoidance learning, scopolamine induced deficit in passive avoidance and reversal of scopolamine deficit by tacrine. PMID- 15882897 TI - Words in the brain's language: an experimental investigation. AB - According to Pulvermuller (1999), words are represented in the brain by cell assemblies (Hebb, 1949) distributed over different areas, depending on semantic properties of the word. For example, a word with strong visual associations will be represented by a cell assembly involving neurons in the visual cortex, while a word suggesting action will selectively activate neurons in the motor areas. The present work aims to test the latter hypothesis by means of behavioural measures. Specifically it tests the prediction that there should be a selective influence (in terms either of interference or priming) of performed/observed movements on the performance (reaction times and accuracy) of lexical decision involving words with a strong action association. Similarly, a selective influence of visual images on lexical decision involving words with strong visual associations should be observed. Two experiments were carried out. Results provided partial support for the hypothesis. PMID- 15882898 TI - Fractographic analyses of three ceramic whole crown restoration failures. AB - OBJECTIVES: Develop fractographic techniques to perform failure analyses of three ceramic whole-crown clinical failures. METHODS: Three clinical whole-crown failures of different ceramic material systems were studied. The systems included appropriate veneers applied to core materials of cold isostatically pressed alumina, injection molded alumina/alumina-magnesia spinel, and hot pressed lithium disilicate. The surface topography was examined for classical fractographic features, utilizing both optical and scanning electron microscopes independently as well as group examination by four fractographers. RESULTS: All three restorations displayed many common fractographic features, the most important of which was wake hackle. Wake hackle occurs when an approaching crack front sweeps by a small bubble or discontinuity in a brittle material, leaving a trail. The resulting trails can be used to trace back crack progression to an initial starting area. Wake hackle was found to be common in the veneer portions of the crowns, and the strategy of creating a wake hackle map enabled crack paths to be traced from a core area outward in all three restorations. The fracture origin areas were located where hoop stresses are high. Another helpful fracture feature was compression curl, an indication of flexural stress, most visible under low incident optical lighting. SIGNIFICANCE: This study indicates that fractographic analyses can be productively performed on ceramic whole-crown failures. Optical examination under low incident lighting and wake hackle mapping were developed as promising techniques. Hoop stress was implicated as a concern in the three studied specimens, and should be considered when analyzing other whole-crown failures. PMID- 15882899 TI - Characterization of hydroxyapatite films obtained by pulsed-laser deposition on Ti and Ti-6AL-4v substrates. AB - OBJECTIVES: Pulsed-laser deposition (PLD) is a development process to obtain hydroxyapatite (HA) thin film. It is an alternative to hydroxyapatite deposition techniques usually employed to cover orthopaedic or dental titanium implant surfaces. The aim of this study is to find out the characteristic ratio for Ca/P (1.66) deposit on titanium implant with the PLD process. METHODS: In a preliminary study, the coating parameters of pure and highly crystalline HA on Ti or Ti-6Al-4V substrates were verified by analysing the deposit by Rutherford backscattering spectroscopy (RBS). Ablation parameters to reach a stoichiometric hydroxyapatite composition (ideal Ca/P atomic ratio) and to control the growth of crystalline phases were: 575 degrees C for the substrate temperature, 0.4 mbar H2O vapour pressure in the ablation chamber, the target substrate distance was 40 mm and the deposition time was 120 min. In a second part, the film properties were analysed by means of XRD, SEM, AFM. The coating adhesion of the HA to the substrate was determined with a micro scratch tester. RESULTS: The analysed HA thin films showed a perfect crystallized and textured deposit. Sample observation and surface quality analysis demonstrated a surface roughness and adhesion of the films to the substrates compatible with biological applications. SIGNIFICANCE: These results suggest that pulsed-laser deposition is a suitable technique to obtain crystalline and adherent hydroxyapatite films on Ti or Ti-6Al-4V substrates. The quality of the HA deposit with the PLD process could be an interesting option for coating dental implant. PMID- 15882900 TI - Novel amino acid modified zinc polycarboxylates for improved dental cements. AB - OBJECTIVE: The objective of this study was to develop a novel amino acid modified zinc/calcium polycarboxylate cement system, formulate the cements, and evaluate their mechanical strengths. MATERIALS AND METHODS: Acrylate and methacrylate derivatives of four amino acids were synthesized and characterized using FT-IR and 1HNMR spectroscopy. The derivatives were formulated with polymer having pendent methacrylate group, water, and synthesized novel filler. Compressive (CS) as well as flexural strengths (FS) and viscosities of the resin liquids were used as tools to evaluate the formulations and formed cements. The specimens for CS and FS tests were conditioned in distilled water at 37 degrees C for 24 h and 7 d, respectively, prior to testing. RESULTS: The measured CS and FS of the cements was in the decreasing order of methacryloyl beta-alanine > acryloyl glutamic acid = acryloyl beta-alanine > methacryloyl glutamic acid > 2-hydroxyethyl methacrylate. Methacryloyl beta-alanine was selected for further formulations due to its relatively low solution viscosity and high CS as well as FS. Effects of polymer content, P/L ratio, tartaric acid and initiator concentration were significant. During aging, the cement showed a constant increase in CS for up to 1 month and then kept constant for up to 3 months. CONCLUSIONS: This study developed a novel amino acid modified zinc/calcium polycarboxylate cement system. This system was 85% higher in CS, 98% higher in DTS and 183% higher in FS, compared to Durelon. PMID- 15882901 TI - Efficient characterisation of human cell-bioceramic interactions in vitro and in vivo by using enhanced GFP-labelled mesenchymal stem cells. AB - Human mesenchymal stem cells (hMSCs) were transfected using four retroviral pseudotypes, amphotropic murine leukemia viruses 4070 (MuLV-10A1), a modification of amphotropic pseudotype 4073 (A71G, Q74K, V139M), gibbon ape leukemia virus (GaLV), or feline endogenous virus (RD114) encoding the neomycin resistance (Neo(r)) gene and enhanced green fluorescent protein (eGFP) as genetic markers. It was observed that the MuLV4073 was the most efficient pseudotype for hMSC transfection. The proliferation and differentiation characteristics of eGFP labelled hMSCs were not significantly different from control hMSCs. G418 selected eGFP-labelled cells were cultured for 3 weeks on two porous, commercially available calcium phosphate bioceramics, a "synthetic hydroxyapatite" and a "deproteinised bone", before implantation into NOD/SCID mice for up to 4 weeks. The eGFP-labelled hMSCs could be readily visualised by their intense green fluorescence both in vitro and in vivo. In "synthetic hydroxyapatite" implants the cells remained in a monolayer, whereas in "deproteinised bone" implants mineralised tissues were detected by histology, scanning electron microscopy and energy dispersive X-ray spectrometry. From the results, it is concluded that the use of eGFP-labelled hMSCs is an effective tool to trace the fate of hMSCs and evaluate the interactions between cells and ceramics both in vitro and in vivo. This is of great value in prospective assessments of these cell populations for use in tissue engineering applications. PMID- 15882902 TI - Estrogen stimulates galanin expression within luteinizing hormone-releasing hormone-immunoreactive (LHRH-i) neurons via estrogen receptor-beta (ERbeta) in the female rat brain. AB - Among the many factors that integrate the activity of the luteinizing hormone releasing hormone (LHRH) neuronal system, estrogens play the most important role. Until recently, the belief has been that the LHRH neurons do not contain estrogen receptors and that the action of estrogen upon LHRH neurons is indirect involving several, estrogen-sensitive neurotransmitter and neuromodulator systems that regulate the activity of the LHRH neurons. Based on our recent findings that LHRH neurons of the female rat co-express galanin, that galanin is a potent LHRH releasing peptide, and that estrogen receptor-beta (ERbeta) is present in LHRH neurons, we have evaluated the effect of 17beta-estradiol on the expression of galanin within LHRH neurons. By combining immunocytochemistry for LHRH and in situ hybridization histochemistry for galanin, we demonstrate that 17beta estradiol stimulates galanin expression within 2h following their administration to ovariectomized rats. Maximal expression, however, required a longer treatment regimen (3 days). These observations strongly suggest that estrogens stimulate galanin expression within LHRH neurons directly, via ERbeta. Moreover, ERbeta may mediate, at least in part, the positive feedback effect of estrogens during the preovulatory LHRH and subsequent LH surges. PMID- 15882903 TI - Ocular conditions associated with peripapillary subretinal neovascularization, their relative frequencies, and associated outcomes. AB - OBJECTIVE: To determine frequency and outcomes of conditions with peripapillary subretinal neovascular membranes (PSRNVMs). DESIGN: Retrospective observational case series. PARTICIPANTS: All patients from a private community-based retina practice diagnosed with a PSRNVM. METHODS: Review of clinical charts, photographs, and fluorescein angiograms of 115 eyes of 96 patients, accrued over 18 years. MAIN OUTCOME MEASURES: Demographics, visual acuity (VA), laterality, neovascular membrane type and recurrence status over treatment course, and associated ocular conditions. RESULTS: Prevalences of reported associations were 52 (45.2%), age-related maculopathy (ARM); 45 (39.1%), idiopathic; 5 (4.3%), multifocal choroiditis; 3 (2.6%), angioid streaks; 2 (1.7%), histoplasmosis; 2 (1.7%), choroidal osteoma; 1 (0.9%), optic disc drusen; and 1 (0.9%), congenital disc anomaly. Newly recognized associations include pattern dystrophy (3 [2.6%]) and peripapillary pseudopodal pigment epithelial and choroidal atrophy (1 [0.9%]). Second-eye involvement was observed in 19.8% of patients over a median follow-up of 2 years. Median initial VAs were 20/40 for ARM-associated eyes and 20/30 for idiopathic eyes (P = 0.0230). Median final VAs were 20/70 for ARM associated eyes and 20/32 for idiopathic eyes (P = 0.0261). The VA changes in the ARM-associated and idiopathic groups did not differ significantly (P = 0.1453). Recurrence of PSRNVMs after laser ablation was seen in 14 of 73 eyes (19.2%). A case of a PSRNVM as a cause of pseudopapilledema leading to unnecessary neurological imaging is reported. CONCLUSIONS: Close inspection of fellow eyes at the time of first eye diagnosis and regular follow-up afterward are indicated, given the high rate of eventual bilateral involvement regardless of associated condition. Laser ablation of PSRNVMs with broad treatment margins reduces recurrence rates relative to earlier series. The differential diagnosis of disc edema should include PSRNVMs. Pattern dystrophy can be associated with PSRNVMs. PMID- 15882904 TI - Redefining the limit of the outer retina in optical coherence tomography scans. AB - OBJECTIVE: A highly reflective layer seen in retinal optical coherence tomography (OCT) has been believed to correspond to the choriocapillaris (CHC) and retinal pigment epithelium (RPE). On gray-scale scans of OCT-2000, and on Stratus OCT, this layer by the outer retinal limit can be resolved into 2 distinct laminae. We analyzed these 2 laminae in normal and abnormal maculae to infer their anatomic correlate. DESIGN: Retrospective study. METHODS: Analysis of macular OCT scans was performed in 44 patients using OCT-2000, and in 39 patients using Stratus OCT. Thirty of these patients had no ocular disease, and their OCT was normal. The other 53 patients had several macular diseases of different etiologies. Both color and gray-scale images were analyzed. RESULTS: Macular OCT scans showed a double laminae at the level where the retina interfaces the RPE in normal subjects using both OCT-2000 and Stratus OCT. In 2-dimensional scans, this laminar structure appears as a double line. It is best distinguished on the Stratus OCT and gray-scale images of OCT-2000. This double line consisted of a thin inner line and a thicker outer line. Similar analysis in patients with macular pathology showed a discernible double line at the retina/RPE interface in at least part of the scan. However, in patients with macular hole, the area corresponding to the absent retina showed only a single line. The inner line component appeared to follow the contour of the retina. This phenomenon was also seen in eyes with neurosensory detachment secondary to central serous chorioretinopathy and other etiologies. In contrast, in macular pathologies where the outer retina did not lose contiguity with the RPE, such as in lamellar macular hole and in cystoid macular edema, the double line persisted. Software for retinal thickness measurements regularly place the outer limit of the retina at the internal aspect of the inner line, probably underestimating the retinal thickness by about 24 to 34 mum. CONCLUSIONS: A double laminar structure at the outer retina/RPE/CHC interface can be consistently distinguished on commercially available OCT of normal eyes. In eyes with macular pathology, OCT analysis of the inner lamina leads us to conclude it is most likely part of the neurosensory retina and not part of the RPE/CHC complex as previously thought. PMID- 15882905 TI - Coats' disease diagnosed in adulthood. AB - PURPOSE: To investigate the manifestations and clinical course of Coats' disease diagnosed in adulthood and to describe the characteristics of patients who develop it. DESIGN: Retrospective observational case series. PARTICIPANTS: Thirteen eyes of 13 patients with Coats' disease diagnosed after age 35 years. METHODS: All patients examined in the authors' referral practices for Coats' disease diagnosed after the age of 35 years were identified. Patients with a history of radiation exposure, intraocular inflammation, retinal vascular occlusion, or Coats' response were excluded. A review of their historical, clinical, and fluorescein angiographic features and demographics was performed. RESULTS: There were 13 patients with a mean age at diagnosis of 50 years (range, 36-79) and a mean follow-up of 5.8 years (range, 0-17). These patients manifested many findings typical of Coats' disease, including the unilateral nature of the disease (13/13 patients), male predominance (12/13), vascular telangiectasis, lipid exudation, macular edema, and areas of capillary nonperfusion with adjacent webs of filigreelike capillaries. Disease was limited to < or =6 clock hours in 10 of 13 patients (76.9%), did not generally involve the retinal vasculature anterior to the equator, and showed limited potential to expand during the follow up period. A localized hemorrhage was noted in 10 of 13 patients (76.9%), almost always near larger aneurysmal dilatations. Limited exudative detachment of the retina was seen in 4 eyes, 3 of which responded to localized laser photocoagulation. One patient had subretinal fluid seen on ocular coherence tomography that did not require treatment. On average, patients lost 2.1 lines of visual acuity during the follow-up period; 6 patients had a decline in vision, 2 patients had improved vision, and 3 patients had stable vision. No patient had end-stage findings of Coats' disease, such as iris neovascularization or total exudative detachment. CONCLUSIONS: Coats' disease can first be diagnosed in adulthood with retinal vascular abnormalities similar to those seen in younger patients. There are a number of important differences in disease manifestation in adults, including limited area of involvement, slower apparent progression of disease, and hemorrhage near larger vascular dilatations. PMID- 15882906 TI - Change in intraocular pressure measurements after LASIK the effect of the refractive correction and the lamellar flap. AB - OBJECTIVE: To study the relationship between intraocular pressure (IOP) readings after LASIK and the amount of refractive correction. DESIGN: Retrospective noninterventional case series. PARTICIPANTS: Patients receiving primary LASIK for myopia and myopic astigmatism. METHODS: A database of preoperative, intraoperative, and 3-month postoperative data for 8113 consecutive eyes that underwent primary myopic and myopic astigmatic LASIK was retrospectively reviewed. Linear regression analysis of measured IOP change as a function of refractive change was then performed. Age and preoperative keratometry were also reviewed by multiple regression. MAIN OUTCOME MEASURES: Best-fit curve relating change in measured IOP to refractive change. RESULTS: The mean spherical equivalent of the refractive change was -4.98+/-2.64 diopters (mean +/- standard deviation). The mean decrease in measured IOP was 2.0+/-3.3 mmHg. Linear regression analysis revealed a decrease of 0.12 mmHg of measured IOP per diopter of refractive change (95% confidence interval [CI], 0.09-0.15, R(2) = 0.009, P<0.001). Extrapolation of the data to a theoretical correction of zero diopters revealed a decrease of 1.36 mmHg (95% CI, 1.20-1.51, P<0.001), suggesting a component of measured IOP change that is independent of laser ablation. CONCLUSIONS: The reduction of IOP readings after corneal refractive surgery is a linear function of the amount of refractive correction, with an additional constant reduction that is probably related to the lamellar corneal flap. These data suggest that the lamellar corneal flap makes no contribution to the load bearing characteristics of the post-LASIK cornea. PMID- 15882907 TI - Pre-excimer laser and post-excimer laser refractive surgery measurements of scotopic pupil diameter using 2 pupillometers. AB - PURPOSE: To compare a digital infrared pupillometer with a handheld light amplification pupillometer for measuring scotopic pupil size and to evaluate if the postoperative refractive changes of the cornea can influence pupil measurements. DESIGN: Prospective noncomparative interventional case series. PARTICIPANTS: One hundred eyes, 50 myopic (mean spherical equivalent [SE] refraction [+/- standard deviation], -4.32+/-2.44 diopters [D]) and 50 hyperopic (mean SE refraction, +2.95+/-0.99 D), of 50 otherwise healthy subjects underwent photorefractive keratectomy or LASIK. INTERVENTION: The preoperative and postoperative scotopic pupil sizes were measured by 2 examiners (E1, E2) with both a handheld light amplification pupillometer (Colvard, Oasis Medical, Glendora, CA) and a digital infrared pupillometer (Eye World Pupillometer [EWP], Oculus Keratograph, Oculus Opikgerate GmbH, Wetzlar, Germany). The agreement and interrater repeatability were determined using the comparison method described by Bland and Altman. The paired Student's t test was used to evaluate the difference between the preoperative and postoperative measurements. MAIN OUTCOME MEASURES: Scotopic pupil diameter, topographic corneal refractive power, uncorrected visual acuity (VA), best spectacle-corrected VA, and manifest spectacle refraction. RESULTS: The preoperative mean scotopic pupil diameter was 6.12+/-0.90 mm with the EWP and 6.18+/-0.91 mm with the Colvard. After the surgery, mean SE refractions were -0.22+/-0.98 D (myopic patients) and +0.19+/-0.40 D (hyperopic patients). Postoperative mean scotopic pupil diameters were 6.12+/-0.89 mm (EWP) and 6.17+/-0.90 mm (Colvard). There was no statistically significant difference between preoperative and postoperative mean scotopic pupil sizes in either patient group. The limits of agreement between the 2 devices ranged from 2.24 mm (E1) to 2.12 mm (E2) preoperatively and from 2.27 mm (E1) to 2.08 mm (E2) postoperatively. The coefficient of interrater repeatability ranged from 0.56 mm (EWP) to 1.12 mm (Colvard) preoperatively and from 0.62 mm (EWP) to 1.14 mm (Colvard) postoperatively. CONCLUSIONS: The digital infrared pupillometer showed better preoperative and postoperative repeatability than the handheld light amplification pupillometer. In the present study, a mean correction of <3 D of the corneal refractive power did not seem to modify the preoperative scotopic pupil size measurements. PMID- 15882908 TI - Macular degeneration in a patient with aceruloplasminemia, a disease associated with retinal iron overload. AB - PURPOSE: To provide the first ophthalmic case report of a Caucasian patient with the rare autosomal recessive disease aceruloplasminemia, which results in iron overload in the retina, brain, and pancreas. DESIGN: Single observational case report. METHODS: Perls' staining of a conjunctival biopsy was used to detect elevated iron levels in the conjunctival epithelium. Fundus photography, fluorescein angiography, and electroretinography were used to document retinal appearance and function. RESULTS: Unlike a report of a Japanese patient with aceruloplasminemia, who had midperipheral retinal pigment epithelium (RPE) cell atrophy and yellowish discoloration of the fundus, our Caucasian patient had a maculopathy. Beginning at age 47, he had development and progression of multiple subretinal yellowish-white lesions and RPE cell atrophy. To confirm tissue iron overload in our patient, we took the novel approach of a conjunctival biopsy, which showed Perls' Prussian blue-positive epithelial cells. CONCLUSIONS: Given our recent finding of elevated iron levels in the RPE of patients with age related macular degeneration (AMD), it is interesting that retinal iron overload in aceruloplasminemia is associated with a maculopathy that clinically resembles AMD. This finding supports the hypothesis that retinal iron homeostasis is essential for normal retinal function. Disruption of iron homeostasis could contribute to the pathogenesis of AMD. PMID- 15882909 TI - Clinical results with the Trabectome for treatment of open-angle glaucoma. AB - OBJECTIVE: To describe clinical results from a pilot study of a novel glaucoma surgical device. DESIGN: Prospective interventional case series. PARTICIPANTS: Thirty-seven adult Hispanic and Caucasian patients (17 male, 20 female) with uncontrolled open-angle glaucoma (OAG) in one or both eyes with or without previous surgery or laser treatment were recruited from a clinical practice in Tijuana, Mexico. INTERVENTION: Surgery was performed with the Trabectome (NeoMedix Corp., San Juan Capistrano, CA) in one eye of each patient. MAIN OUTCOME MEASURES: Goldmann applanation intraocular pressures and Snellen visual acuities were measured before and after surgery. Intraoperative and postoperative adverse events were tabulated, and numbers of preoperative and postoperative adjunctive medications were compared before and after surgery. RESULTS: Preoperative pressures after 1 week of medication washout averaged 28.2+/-4.4 mmHg (n = 37). Only 3 patients were not using topical medications preoperatively. Follow-up ranged between 3 months (n = 37) and 13 months (n = 11). Mean postoperative IOPs were 18.4+/-10.9 mmHg (n = 37) at 1 day, 17.5+/-5.9 mmHg (n = 37) at 1 week, 17.4+/-3.5 mmHg (n = 25) at 6 months, and 16.3+/-2.0 mmHg (n = 15) at 12 months. Visions returned to within 2 lines of preoperative levels and remained stable in all patients beyond 3 weeks postoperatively except one, not sutured at surgery, who had a late hyphema probably associated with corneal wound gaping after accidental blunt trauma. The number of adjunctive medications decreased from 1.2+/-0.6 among preoperative patients on medications (n = 34) to 0.4+/-0.6 among all patients at 6 months (n = 25). Blood reflux occurred in all eyes on instrument withdrawal after angle surgery and was present at day 1 in 22 eyes (59%) with clearing by slit-lamp examination at a mean of 6.4+/-4.1 days postoperatively. CONCLUSIONS: The Trabectome seems to offer a safe and effective method of lowering IOP in OAG. PMID- 15882910 TI - Patterns of afferent projections to the dentate gyrus studied in organotypic co cultures. AB - Cholinergic axons originating from the septum form a characteristic layer of preterminal axons and apparent termination in the molecular layer of the hippocampal dentate gyrus. The present study explored the specificity of this characteristic axonal pattern, through the use of organotypic slice co-cultures. Slices of hippocampus were co-cultured with a slice from one of a variety of other potential sources of afferents, and the afferent axons were labeled histochemically or immunocytochemically to determine which afferents distribute within the dentate molecular layer in a pattern similar to that formed by septal cholinergic projections. Acetylcholinesterase (AChE) histochemistry demonstrated that cholinergic axons from septum, substantia innominata, and striatum all consistently targeted the inner molecular layer of the dentate gyrus. AChE labeled cholinergic axons from dorsal lateral pontine tegmentum and from spinal cord sometimes formed this pattern, while axons from the habenula failed to extend into the dentate gyrus. Immunocytochemically identified monoaminergic axons from the substantia nigra, locus coeruleus, and raphe extended into co cultured hippocampus; each of these afferent systems displayed a prominent axonal plexus within the hilus of the dentate, but only the raphe axons projected prominently to the molecular layer. These data demonstrate that the molecular layer of the dentate gyrus provides an attractive target zone for some cholinergic and monoaminergic afferents, but not all. Commonalities between neuronal populations that preferentially project to the molecular layer in vitro may offer clues regarding the axon guidance mechanisms that normally direct cholinergic axons to target sites in the dentate gyrus molecular layer. PMID- 15882911 TI - Extensive expression studies revealed a complex alternative splicing pattern of the HMGA2 gene. AB - Chromosomal rearrangements of the HMGA2 locus belong to the most common aberrations in human benign tumors. HMGA2 rearrangements often result in chimeric genes expressing transcripts consisting of the first three exons of HMGA2 followed by ectopic sequences derived from intron 3 of that gene. RT-PCR-based expression studies of 4 of these HMGA2 transcripts revealed a co-expression with the "wild-type" HMGA2a in tumor samples as well as in normal tissues. Northern blot hybridizations of the lipoma cell line Li-14 revealed the expression of five additional HMGA2 transcripts consisting of exons 1 to 3 but not exons 4 to 5 besides the full-length HMGA2a transcript. In silico analyses have been performed showing a high homology to well-established consensus sequences for the 3' splice acceptor site, the branch site, and poly(A) signal. Thus, it is quite obvious that the HMGA2 transcripts described herein are alternative, not aberrant, splice products of the HMGA2 gene. It is hypothesized that HMGA2-dependent tumorigenesis is caused by a disturbed equilibrium in the co-expression of the HMGA2 splice variants leading to aberrant cell proliferation and/or malignant transformation of cells. PMID- 15882912 TI - Cardiovascular reactivity during dyadic social interaction: the roles of gender and dominance. AB - The present study examined relationships among gender, trait dominance, and cardiovascular reactivity assessed during dyadic social interactions between same gender and same-ethnicity participants. One-hundred sixty dyads, 76 comprised of African American participants and 84 comprised of European American participants, engaged in three 4-min discussions while their heart rate and blood pressure responses were assessed. All dyads were composed of unacquainted partners. Multilevel modeling for dyadic data revealed that trait dominance was significantly and positively associated with blood pressure reactivity during the discussions, but only among men who interacted with a highly dominant partner. The results of the present study add to the weight of the evidence identifying dominance as a social domain of significance for cardiovascular functioning, particularly among men. Potential reasons for gender-specific patterning are considered. PMID- 15882913 TI - The KPNA3 gene may be a susceptibility candidate for schizophrenia. AB - The present study investigated the possible association of the KPNA3 locus in the 13q14 region with schizophrenia. We detected 7 single nucleotide polymorphisms (SNPs) on 13q14, one (rs6313) present at the HTR2A locus and the other 6 at the KPNA3 locus, among 124 British family trios consisting of mother, father and affected offspring with schizophrenia. The transmission disequilibrium test (TDT) showed allelic association for rs3736830 (chi(2)=8.66, P=0.003), rs2181185 (chi(2)=3.86, P=0.049) and rs626716 (chi(2)=5.82, P=0.016), but not for rs6313 (chi(2)=0.009, P=0.926). The global P-value was 0.029 for 1000 permutations with the TDT. The 2-SNP haplotype analysis showed a disease association for the rs2273816-rs3736830 haplotypes (chi(2)=7.63, d.f.=2, P=0.022), the rs3736830 rs2181185 haplotypes (chi(2)=10.30, d.f.=2, P=0.006) and the rs2181185-rs3782929 haplotypes (chi(2)=9.26, d.f.=2, P=0.01). The global P-value was 0.034 for 1000 permutations with the 2-SNP haplotype analysis. The 6-SNP haplotype system also showed a weak association with the illness (chi(2)=15.62, d.f.=8, P=0.048), although the 1-d.f. test did not show the association for nine individual haplotypes when a P-value was corrected by the Bonferroni corrections. The present study suggests that the KPNA3 may contribute genetically to schizophrenia in a small effect size. PMID- 15882915 TI - (137)Cs concentrations in Atlantic and western Antarctic surface waters: results of the 7th Ukrainian Antarctic Expedition, 2002. AB - The latitudinal distribution of (137)Cs in the Atlantic--western Antarctic surface waters was studied during the 7th Ukrainian Antarctic Expedition in January--May 2002. The (137)Cs concentrations have also been measured in the upper ice of the coastal glacier Woozle Hill located near the Ukrainian Antarctic station "Akademik Vernadsky" (western Antarctica, 65 degrees 15' S-64 degrees 16' W). Comparison of these data with results of previous same-route expeditions SWEDARP (Swedish Antarctic Research Expedition, 1988/1989) and the French R/V "Jeanne d'Arc" (1992/1993), has shown practically parallel changes of (137)Cs surface concentrations between 40 degrees N and 20 degrees S, pointing to decrease of (137)Cs radioactivity at these latitudes with an apparent half-life of 10--15 years (12.5+/-2.1 years on average). This suggests that decrease of (137)Cs surface concentration within this latitude band is controlled, besides the radioactive decay of (137)Cs (half-life=30 years), by vertical mixing of the upper water masses. South of 20 degrees S, the (137)Cs concentrations in surface water have decreased more rapidly because of the influence of the less contaminated Antarctic waters. At 50--60 degrees S and near the Antarctic coast, the (137)Cs activity in 2002 was similar to those measured during the SWEDARP and "Jeanne d'Arc" expeditions, suggesting an additional input of (137)Cs to these waters from the melted ice from the adjacent glaciers. PMID- 15882914 TI - Pituitary adenylate cyclase-activating polypeptide (PACAP) mimics neuroendocrine and behavioral manifestations of stress: Evidence for PKA-mediated expression of the corticotropin-releasing hormone (CRH) gene. AB - The physiologic response to stress is highly dependent on the activation of corticotropin-releasing hormone (CRH) neurons by various neurotransmitters. A particularly rich innervation of hypophysiotropic CRH neurons has been detected by nerve fibers containing the neuropeptide PACAP, a potent activator of the cAMP protein kinase A (PKA) system. Intracerebroventricular (icv) injections of PACAP also elevate steady-state CRH mRNA levels in the paraventricular nucleus (PVN), but it is not known whether PACAP effects can be associated with acute stress responses. Likewise, in cell culture studies, pharmacologic activation of the PKA system has stimulated CRH gene promoter activity through an identified cAMP response element (CRE); however, a direct link between PACAP and CRH promoter activity has not been established. In our present study, icv injection of 150 or 300 pmol PACAP resulted in robust phosphorylation of the transcription factor CREB in the majority of PVN CRH neurons at 15 to 30 min post-injection and induced nuclear Fos labeling at 90 min. Simultaneously, plasma corticosterone concentrations were elevated in PACAP-injected animals, and significant increases were observed in face washing, body grooming, rearing and wet-dog shakes behaviors. We investigated the effect of PACAP on human CRH promoter activity in alphaT3-1 cells, a PACAP-receptor expressing cell line. Cells were transiently transfected with a chloramphenicol acetyltransferase (CAT) reporter vector containing region - 663/+124 of the human CRH gene promoter then treated for with PACAP (100 nM) or with the adenylate cyclase activating agent, forskolin (2.5 muM). Both PACAP and forskolin significantly increased wild-type hCRH promoter activity relative to vehicle controls. The PACAP response was abolished in the CRE-mutant construct. Pretreatment of transfected cells with the PKA blocker, H 89, completely prevented both PACAP- and forskolin-induced increases in CRH promoter activity. Furthermore, CREB overexpression strongly enhanced PACAP mediated stimulation of hCRH promoter activity, an effect which was also lost with mutation of the CRE. Thus, we demonstrate that icv PACAP administration to rats under non-stressed handling conditions leads to cellular, hormonal and behavioral responses recapitulating manifestations of the acute stress response. Both in vivo and in vitro data point to the importance of PACAP-mediated activation of the cAMP/PKA signaling pathway for stimulation of CRH gene transcription, likely via the CRE. PMID- 15882916 TI - The effect of pes cavus on foot pain and plantar pressure. AB - BACKGROUND: Clinical management of patients with painful pes cavus is challenging because the mechanism of foot pain is poorly understood. The purpose of this study was to explore the influence of various pes cavus aetiologies on foot pain and plantar pressure characteristics, and to identify the relationship between foot pain and plantar pressure. METHODS: Seventy subjects were recruited for this study. They included 30 subjects with pes cavus of unknown aetiology (idiopathic), 10 subjects with pes cavus of neurological aetiology (neurogenic) and 30 subjects with a normal foot type. The presence and location of foot pain was recorded and barefoot plantar pressures were measured using the EMED-SF platform for the whole foot, rearfoot, midfoot and forefoot regions. FINDINGS: Subjects with pes cavus of either idiopathic or neurogenic aetiology reported a higher proportion of foot pain (60%) compared to subjects with a normal foot type (23%) (P=0.009). Pressure-time integrals under the whole foot, rearfoot and forefoot regions in pes cavus, of both idiopathic and neurogenic origin, were higher than in the normal foot type (P<0.01). Pressure-time integrals in subjects reporting foot pain were higher than for pain free subjects (P<0.001). There was a significant correlation between pressure-time integral and foot pain (r=0.49, P<0.001). INTERPRETATION: Foot pain is a common finding among individuals with pes cavus. Regardless of aetiology, pes cavus is characterized by abnormally high pressure-time integrals which are significantly related to foot pain. An understanding of the relationship between pes cavus pressure patterns and foot pain will improve the clinical management of these patients. PMID- 15882917 TI - A comparison of the rhizosphere-based method with DTPA, EDTA, CaCl2, and NaNO3 extraction methods for prediction of bioavailability of metals in soil to barley. AB - A rhizosphere-based method was compared with DTPA, EDTA, CaCl2, and NaNO3 extraction methods for the evaluation of bioavailability of heavy metals in soil to barley. The extractable amounts of Cr, Cu, Zn and Cd analyzed by the rhizosphere-based method correlated significantly with the metal content of barley roots. The extractable metals identified by DTPA, EDTA, CaCl2 and NaNO3 methods exhibited relatively poor or no correlation with the metal content of barley roots. The stepwise multiple regression equation of the rhizosphere-based method was the simplest one, as no soil properties needed to be entered, whereas the equations for the DTPA, EDTA, CaCl2 and NaNO3 extraction methods always require those variables. The most distinct feature of the rhizosphere-based method was that the proposed method was suitable for acidic, neutral and near alkaline soils. In contrast, the other extraction methods were restricted to soil types. In summary, the rhizosphere-based method is the most robust approach for evaluation of bioavailability of metals in soil to barley. PMID- 15882918 TI - Management options to reduce exposure to methyl mercury through the consumption of fish and fishery products by the French population. AB - This paper presents an updated assessment of exposure in France to methyl mercury through the consumption of fish and fishery products, and proposes several management scenarios which could reduce this exposure through changes to fish contamination levels or fish consumption patterns. The exposure model was applied in line with previous methodological results [Tressou, J., Crepet, A., Bertail, P., Feinberg, M.H., Leblanc J.Ch., 2004a. Probabilistic exposure assessment to food chemicals based on extreme value theory: application to heavy metals from fish and sea products. Food Chem. Toxicol. 42, 1349-1358; Tressou, J., Leblanc, J.Ch., Feinberg, M., Bertail, P., 2004b. Statistical methodology to evaluate food exposure to a contaminant and influence of sanitary limits: application to ochratoxin A. Regul. Toxicol. Pharmacol. 40, 252-263] so as to obtain a realistic estimate of probability and confidence intervals (95% CI) concerning French consumers exposed to levels exceeding the revised fixed provisional tolerable weekly intake (PTWI) for methyl mercury of 1.6 microg/week/kg of body weight, established by the Joint FAO/WHO Expert Committee on Food Additives in 2003. The results showed that young children aged between 3 and 6 years old or 7 and 10 years old, and women of childbearing age were at the risk groups. With respect to these groups and according to the fish consumers patterns (consumers of predatory fish only or consumers of predatory and nonpredatory fish), the results suggested that strategies to diminish MeHg exposure by reducing the amount of predatory fish consumed would be more efficient in significantly decreasing the probability of exceeding the PTWI than the implementation of international standards. PMID- 15882919 TI - Reluctant acceptance of xenotransplantation in kidney patients on the waiting list for transplantation. AB - Previous research has shown contradictory results regarding the acceptance of xenotransplantation (XT) by kidney patients. This variance in attitude towards XT could be explained by differences in knowledge of XT, the availability of alternative treatment options, specific beliefs and cognitions of the patient, and differences in the health status of the patients. We studied 61 patients in this contention in the Netherlands waiting for a kidney transplant. All took part in in-depth interviews and filled out questionnaires on two occasions, once before and once after an information brochure on XT had been read. We found that a better health status is correlated with a greater acceptance of XT, but only before information was given. After information had been given, the acceptance of XT decreased significantly. However, if XT is the only possibility in a life threatening situation, almost all patients are willing to accept XT, except for a small group (approximately 10-15%) with fundamental religious and/or spiritual objections against such treatment. When alternative treatment options such as a human cadaver donor, a living related donor or commercial donor are offered, the majority of patients would prefer those to XT. The main reason for reluctance is uncertainty about the risks of XT to personal health. Because alternatives are currently available, we expect that XT will become more popular only if future results of this procedure have proved to be comparable to transplantation with a human donor kidney. PMID- 15882920 TI - Physicians' willingness to report impaired colleagues. AB - We surveyed physicians to determine what factors were associated with their reporting of impaired colleagues to Physician Health Programs (PHPs). We conducted a cross-sectional mail survey of 1000 randomly selected practicing physicians in the United States. A survey instrument asked the physicians whether they would report 10 hypothetical impaired colleagues to a PHP. The results show that a majority of the physicians would report physicians to PHPs, but were more likely to report hypothetical physicians involved in substance abuse than those who were emotionally or cognitively impaired (p<0.001). Respondents who felt they had a societal obligation as opposed to an obligation to protect the rights of the individual (p=0.006) were more likely to report hypothetical physicians. Those respondents who stated they knew of guidelines on reporting impaired physicians had more frequently reported impaired colleagues (p<0.001). We conclude that physicians should be educated on the availability and functioning of PHPs and the ethical and legal obligations of assisting impaired colleagues. PMID- 15882921 TI - Establishing visual category boundaries between objects: a PET study. AB - Individuals with Alzheimer's disease (AD) often have problems in recognizing common objects. This visual agnosia may stem from difficulties in establishing appropriate visual boundaries between visually similar objects. In support of this hypothesis, showed that AD subjects have difficulties in establishing visual category boundaries between continuously graded shapes. In an attempt to investigate the neural basis of these impairments, the current study required a group of neurologically healthy elderly participants to categorically classify a series of ellipses varying in width while regional blood flow changes were measured using positron emission tomography (PET). Two categorization conditions were compared in order to isolate changes in cortical activity that dissociated the categorization of shapes situated either near or far from a category boundary that divided the width continuum. The participants produced a discontinuity in the probability and speed of categorizing the shapes at some point along the continuum, suggesting that the objects were classified into distinct categories. Moreover, a comparison of the PET scans obtained while the subjects were categorizing the shapes situated near vs. far form the category boundary revealed significant differences in cortical activity in the parietal and frontal brain areas. These findings suggest that both visuo-spatial and decision making mechanisms may be involved in establishing categorical distinctions between continuously graded stimuli. It is proposed that the functional role of the parietal and frontal cortical regions in establishing visual boundaries between categories of objects may be relevant for understanding object recognition impairments in AD. PMID- 15882922 TI - Bioluminescence regenerative cycle (BRC) system: theoretical considerations for nucleic acid quantification assays. AB - A novel application of bioluminescence for nucleic acid quantification, the bioluminescence regenerative cycle (BRC), is described in theoretical terms and supported by preliminary experimental data. In the BRC system, pyrophosphate (PPi) molecules are released during biopolymerization and are counted and correlated to DNA copy number. The enzymes ATP-sulfurylase and firefly luciferase are employed to generate photons quantitatively from PPi. Enzymatic unity-gain positive feedback is implemented to amplify photon generation and to compensate for decay in light intensity by self-regulation. The cumulative total of photons can be orders of magnitude higher than in typical chemiluminescent processes. A system level theoretical model is developed, taking into account the kinetics of the regenerative cycle, contamination, and detector noise. Data and simulations show that the photon generation process achieves steady state for the time range of experimental measurements. Based on chain reaction theory, computations show that BRC is very sensitive to variations in the efficiencies of the chemical reactions involved and less sensitive to variations in the quantum yield of the process. We show that BRC can detect attomolar quantities of DNA (10(-18) mol), and that the useful dynamic range is five orders of magnitude. Sensitivity is not constrained by detector performance but rather by background bioluminescence caused by contamination by either PPi or ATP (adenosine triphosphate). PMID- 15882923 TI - Local rigidity of a protein molecule. AB - Distribution of soft and rigid substructures within a protein molecule has been implicated in several occasions and most recently from the imaging and indentation experiments using an atomic force microscope. In this paper, previously reported result of mechanical extension experiments on the recombinant bovine carbonic anhydrase II, Q253C, is re-analyzed to estimate the distribution of Young's modulus, Y, in this protein. The force vs. extension curve of the enzymatically active, type I conformer gave an estimate of Y increasing from 40 to 220 MPa as the polypeptide chain was extended from 10 to 75 nm indicating the presence of a rigid core structure. The enzymatically inactive type II, in contrast, gave an almost constant modulus of 55+/-15 MPa in the same extension range in agreement with the previous proposal that it lacked a core structure. PMID- 15882924 TI - Correction of malignant behavior of tumor cells by traditional Chinese herb medicine through a restoration of p53. AB - We have previously demonstrated that a UVC-induced tumorigenic HeLa x skin fibroblast cell line could be induced to form a more normal phenotypic state ('reversion'), including loss of IAP expression. We have now used the loss of IAP expression to monitor the enhancement of this reversion in the cervical cancer cell line, HeLa, by a traditional Chinese herb medicine (TCM), Yigan Kang (YGK). IAP level decreased, and the reversion frequency increased, in a dose-dependent manner at concentrations of YGK of more than 10 mg. YGK significantly repressed E6/E7 oncogenes at the transcriptional level, with subsequent reactivation of p53 and p21 expression (P<0.01). YGK had little effect on the cell cycle of HeLa cells and slightly increased the apoptotic cell death between 20 and 40 mg. In vivo, tumorigenicity studies were performed using six different animal experimental protocols, which demonstrated that YGK was effective at inducing reversion of the tumorigenic phenotype, with YGK-treated HeLa cells showing much less aggressive tumor growth than untreated cells. YGK may raise the possibility of the continuing management of some cancers as a chronic condition in which the malignant behavior of the tumor cells is constrained. PMID- 15882925 TI - A genetic polymorphism in prostaglandin synthase 2 (8473, T-->C) and the risk of lung cancer. AB - Prostaglandin synthase 2 (PTGS2) mediated production of prostaglandins is an important step in the inflammatory response. In a population-based case-cohort study, we investigated the relationship between lung cancer and a polymorphism in PTGS2 at site 8473. Among 54,220 cohort members, 265 lung cancer cases and a comparison group of 272 individuals were identified. We found no overall significant association between the PTGS2 polymorphism and lung cancer risk, though the results indicated a lower risk in subjects carrying one copy (RR 0.87; 95% CI 0.61-1.25) or two copies (RR 0.58; 95% CI 0.30-1.12) of the variant allele compared with subjects carrying two copies of the wildtype allele. This tendency seemed to be strongest among the youngest subjects (50-55 years) with a RR on 0.48 (0.19-1.22) in carriers of one variant allele and 0.21 (0.04-1.07) in carriers of two variant alleles, and a significant trend (P=0.04). These results indicate that PTGS2 could be implicated in the development of lung cancer. PMID- 15882926 TI - Increased expression of connexin 26 in the invasive component of lung squamous cell carcinoma: significant correlation with poor prognosis. AB - Reduced expression of connexins (Cxs), gap junction proteins, is frequently reported in malignant cell lines and tumors, whereas recent studies suggested that C x 26, a subtype of Cxs, might help tumor cells acquire malignant phenotypes. To examine this suggestion in the clinical setting, 50 lung squamous cell carcinomas (SCCs) were stained with the anti-C x 26 antibody. No C x 26 specific signals were detectable in 34 tumors (group I; 68%), whereas the remaining 16 were judged positive for C x 26 (group II; 32%). In 14 tumors of group II, C x 26-specific signals were detected not in all SCC cells but in SCC cells facing the tumor stroma or capsule, in which the signals were localized on the plasma membrane. Involved lymph nodes of group-II patients often contained metastatic foci consisting of all C x 26-positive cells. The proportion of C x 26 positive to C x 26-negative SCC cells in the metastatic nodes was larger than that in the corresponding primary tumors. C x 26-positive SCC cells seemed to be more invasive and metastatic than negative ones. Consistently, the 5-year cancer specific survival rate of group-II patients was significantly lower than that of group-I patients (12.5 vs 38.9%; P=0.0391). Multivariate analysis demonstrated that C x 26 expression (P=0.0448) as well as pathological stage (P=0.0338) and vascular invasion (P=0.0191) were independent, significant prognostic predictors. These results suggest that C x 26 may represent an essential effector for controlling the biological aggressiveness of lung SCC tumor. PMID- 15882927 TI - Induction of gastric GIST in rat and establishment of GIST cell line. AB - Continuous administration of the direct-reacting carcinogen N-methyl-N'-nitro-N nitrosoguanidine reportedly produces not only adenocarcinoma in rats, but also mesenchymal tumors. A large number of tumors diagnosed as gastrointestinal smooth muscle tumors actually represent gastrointestinal stromal tumors (GISTs) in human cases. We have induced mesenchymal tumors by duodenal reflux in rats. To clarify the differentiation of these mesenchymal tumors, immunohistochemical investigations were undertaken. In addition, the first culture model of GIST-DR derived from GIST induced by duodenal reflux was established. GIST-DR cells, both in vitro and in vivo, were strongly immunopositive for both KIT and CD34, and STI571 (Imatinib mesylate) blocked the proliferation of this cell line. The present results suggest that duodenal reflux plays a role in the histogenesis of GIST. PMID- 15882928 TI - Elevated 12- and 20-hydroxyeicosatetraenoic acid in urine of patients with prostatic diseases. AB - The role of eicosanoids (metabolites of arachidonic acid) in prostate diseases is receiving increased attention. We investigated the relationship between the concentrations of urinary free acids of 12- and 20-hydroxyeicosatetraenoic acids (12- and 20-HETE) and the benign prostatic hypertrophy (BPH) and prostate cancer (Pca). Urinary concentrations of 12-HETE and 20-HETE of BPH and Pca patients were significantly higher than normal subjects. After removal of the prostate gland, the urinary concentrations of these eicosanoids decreased to concentrations similar to the normal subjects. These results suggest that urinary free acids of 12-HETE and 20-HETE indicate an abnormality of the prostate gland. PMID- 15882929 TI - WITHDRAWN: Loss of heterozygosity analyses of esophageal squamous cell carcinoma and precursor lesions from a high incidence area in China. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 15882930 TI - Long-term depression is not modulated by ATP receptors in the rat CA1 hippocampal region. AB - ATP is an important extracellular messenger in the CNS. In the hippocampus, a brain structure relevant for learning and memory processes, it acts both as a modulator and as a mediator of synaptic transmission, with implications for synaptic plasticity phenomena. Recent evidence suggests that ATP modulates activity-dependent long-term potentiation (LTP) of Schaffer collateral-CA1 synapses. However, it remains unclear if ATP also modulates LTP counterpart's phenomenon, long-term depression (LTD), in the rat hippocampus. This study investigated the effect of ATP analogues on homosynaptic LTD, induced by low frequency stimulation of the Schaffer collaterals (1 Hz; 900 pulses) in the CA1 region of young rat hippocampal slices. The metabolically stable ATP analogues beta,gamma-ImATP (20 microM), a P2 receptor agonist, and alpha,beta-MeATP (20 microM), a preferential P2X(1,3) receptor agonist, did not modify LTD (LTD values of 14.7+/-0.5% and 14.1+/-3% for aCSF controls and of 15.1+/-4% and 19.0+/-5.2% for beta,gamma-ImATP and alpha,beta-MeATP, respectively). The ATP analogue beta,gamma-ImATP (20 microM) did not modify LTD also in the presence of the adenosine A1 receptor antagonist DPCPX (50 nM) (21.5+/-4.2% for DPCPX only and of 23.8+/-8.9% for DPCPX plus beta,gamma-ImATP). Finally, the preferential P2X(1,3) receptor antagonist NF023 (10 microM) had also no effect on LTD (18.6+/-5.2% for aCSF and of 18.7+/-5.2% for NF023). The present results suggest that ATP does not modulate activity-dependent homosynaptic LTD in the rat CA1 hippocampal region by activating P2 receptors. PMID- 15882931 TI - Acute repetitive transcranial magnetic stimulation reactivates dopaminergic system in lesion rats. AB - Repetitive transcranial magnetic stimulation (rTMS) offers potential benefit as a therapeutic treatment for neurological and psychiatric disorders. However, the mechanism underlying the therapeutic effects of rTMS is still unknown. In this study, we investigated the rescue effects of rTMS in the lesioned rats by administering the neurotoxin MPTP (l-methyl-4-phenyl-l,2,3,6-tetrahydropyridine). The rats received rTMS (10 trains of 25 pulses/s for 8 s) 48 h after MPTP injection, and tyrosine hydroxylase (TH) and NeuN expressions were investigated in the substantia nigra. The functional observational battery-hunched posture score for the MPTP-rTMS group was significantly lower and the number of rearing events was higher compared with the MPTP-sham group, these behavioral parameters revert to control levels. These results suggest that rTMS treatment reactivates the dopaminergic system in lesion rats. PMID- 15882932 TI - Substrate effects on oscillations in metabolism, calcium and secretion in single mouse islets of Langerhans. AB - Glucose induces complex patterns of oscillations in intracellular Ca2+ concentration ([Ca2+]i), metabolism and secretion in islets of Langerhans including "slow" and "fast" pulses with period of 2-5 min and 10-20 s respectively. In an effort to elucidate the origin of slow oscillations, individual mouse islets were exposed to different fuels including glyceraldehyde, pyruvate, methyl pyruvate and alpha-ketoisocaproate (KIC), all of which bypass key steps of glycolytic metabolism, while monitoring [Ca2+]i, oxygen consumption and secretion. Glyceraldehyde gave rise to slow oscillations only when substimulatory glucose was also added to the media. Glucosamine, an inhibitor of glucokinase, blocked these slow oscillations. KIC, pyruvate, and methyl pyruvate did not give rise to slow oscillations alone or with glucose present. The addition of glucose to islets bathed in nutrient-rich cell culture media accelerated metabolism and initiated slow oscillations while glyceraldehyde did not. It is concluded that glucose has a special role in accelerating metabolism and generating slow oscillations in isolated islets of Langerhans from mice. Combined with previous observations of Ca2+ dependency for all oscillations in islets, we propose that interactions between Ca2+ influx and glycolysis are responsible for the slow oscillations. In contrast, fast oscillations can occur independent of glycolytic flux. PMID- 15882933 TI - Adding irinotecan to first-line gemcitabine improves tumour response in advanced pancreatic cancer. PMID- 15882934 TI - Progression in intima media thickness--the significance of hormonal biomarkers of chronic stress. AB - OBJECTIVE: The pathophysiological pathways from stress caused by psychosocial stress to IHD has not been dealt with very extensively. The objective of this study was to analyse the association between cortisol levels and progression in intima media thickness (IMT). METHODS AND RESULTS: In 1998 and 2002, 95 participants went through a clinical investigation including ultrasound of the artery carotis communis. Progression in IMT was analysed in relation to levels of salivary cortisol in 1998 and the average levels of salivary cortisol in 1998/2002. Further, the significance of conventional coronary risk factors, testosterone and dehydro-epiandrosterone sulphate (DHEAS) were evaluated. Among the men, only age and HDL-cholesterol (negative) were significantly correlated with progression in IMT. Among the women, awakening cortisol response was significantly correlated with progression in IMT. Testosterone and DHEAS were borderline significantly associated (negatively) with progression in IMT in both genders. CONCLUSION: Progression in atherosclerosis were determined by different risk factors in women and men. The awakening cortisol response was of great importance to IMT progression in women but not in men. PMID- 15882935 TI - Diagnosis and treatment of neonatal hypotension outside the transitional period. AB - The etiology and pathophysiology of the circulatory compromise are among the primary determinants of the clinical presentation of patients with neonatal shock. Therefore, in the absence of direct assessment of cardiac output and organ blood flow, the characteristic clinical presentation itself may guide the initial management of the circulatory compromise. This chapter discusses different pathophysiology-driven management approaches to a number of characteristic clinical presentations of neonatal shock. The clinical presentations discussed in detail are the hypotensive very low birth weight neonate with a hemodynamically significant patent ductus arteriosus, and the preterm or term neonate with perinatal depression, pressor/inotrope resistance and relative adrenal insufficiency, and with specific systemic inflammatory response syndrome. In the absence of information from appropriately designed, randomized clinical trials, management of neonatal shock remains based on pathophysiology and experience. Thus, as there is little evidence for the effectiveness of these management approaches to improve mortality and short-and long-term outcome, the therapeutic approaches described in this chapter should be carefully evaluated and cautiously entertained when treating a neonate with circulatory compromise. PMID- 15882936 TI - Passiflora actinia Hooker extracts and fractions induce catalepsy in mice. AB - Leaves from several Passiflora species are largely employed in the Brazilian folk medicine for its anxiolytic and sedative properties. In behavioral studies, to analyze the tranquillizer action of Passiflora actinia Hooker, it was noteworthy that animals treated with the hydroethanol (HE) and methanol (ME) extracts presented an abnormal postural immobility compared to control animals. That observation led to an investigation of the effects of HE and its fractions on evaluation of catalepsy in mice. The results showed that HE extract, methanol extract, the sequent aqueous crude fractions (AF), and fa, fb and fc chromatographic fractions obtained from Passiflora actinia induced catalepsy in mice. Apparently, the active principles responsible for catalepsy are present in all of the fractions of the extract. PMID- 15882937 TI - Imaging of the pediatric hand: soft tissue abnormalities. AB - This review attempts to demonstrate the spectrum of soft tissue masses encountered in the child's hand. The diagnosis may be suggested by the combination of physical exam and radiographs. Frequently the etiology of a soft tissue mass cannot be established by conventional radiographs and further imaging is necessary. MRI by virtue of its ability to provide detailed soft tissue characterization is the preferred method of imaging the problematic soft tissue mass. PMID- 15882939 TI - Pleiotropic effects of lactate dehydrogenase inactivation in Lactobacillus casei. AB - In lactic acid bacteria, conversion of pyruvic to lactic acid through the activity of lactate dehydrogenase (Ldh) constitutes the final step of the homofermentative pathway. Lactobacillus casei has two characterized genes encoding Ldh activities. The ldhL gene codes for an L-Ldh, which specifically catalyzes the formation of L-lactate, whereas the hicD gene codes for a D hydroxyisocaproate dehydrogenase (HicDH), which catalyzes the conversion of pyruvate into D-lactate. In L. casei cells fermenting glucose, a mixture of L-/D lactate with a 97:3% ratio was formed. Inactivation of hicD led to undetectable D lactate levels after glucose fermentation, while L-lactate levels remained constant. Inactivation of ldhL did not abolish the production of L-lactate, but the lactate final concentration decreased about 25% compared to the wild type, suggesting the presence of at least a second L-Ldh. Moreover, part of the pyruvate flux was rerouted and half of the lactate produced was in the D-isomer form. ldhL inactivation in L. casei showed additional interesting effects. First, the glycolytic flux from pyruvate to lactate was redirected and other fermentation products, including acetate, acetoin, pyruvate, ethanol, diacetyl, mannitol and CO(2), were produced. Second, a lack of carbon catabolite repression of lactose metabolism and N-acetyl-glucosaminidase activity was observed. This second effect could be partly avoided by growing the cells under aeration, since NADH oxidases could account for NAD+ regeneration. PMID- 15882940 TI - RAGE and amyloid beta interactions: atomic force microscopy and molecular modeling. AB - In the AD brain, there are elevated amounts of soluble and insoluble Abeta peptides which enhance the expression of membrane bound and soluble receptor for advanced glycation end products (RAGE). The binding of soluble Abeta to soluble RAGE inhibits further aggregation of Abeta peptides, while membrane bound RAGE Abeta interactions elicit activation of the NF-kappaB transcription factor promoting sustained chronic neuroinflammation. Atomic force microscopy observations demonstrated that the N-terminal domain of RAGE, by interacting with Abeta, is a powerful inhibitor of Abeta polymerization even at prolonged periods of incubation. Hence, the potential RAGE-Abeta structural interactions were further explored utilizing a series of computational chemistry algorithms. Our modeling suggests that a soluble dimeric RAGE assembly creates a positively charged well into which the negative charges of the N-terminal domain of dimeric Abeta dock. PMID- 15882941 TI - Quantitative assessment of emphysema distribution in smokers and patients with alpha1-antitrypsin deficiency. AB - INTRODUCTION: Identification of upper lobe emphysema is mandatory before lung volume reduction surgery (LVRS). Here we introduce a CT-based objective model for describing the distribution of different types of emphysema. METHODS: Fifty COPD patients were included in the study. Half had alpha1-antitrypsin deficiency (alpha1-COPD) and the rest had smoking-induced emphysema (usual COPD). All patients were scanned 3 times. The relative area of emphysema in each CT slice was plotted against table position, and the cranio-caudal distribution was calculated as the slope of the regression line. RESULTS: The variation in slopes within a patient was much less than the variation in slopes between patients (P<0.0001). There was a significant difference between slopes in the alpha1-COPD and the usual COPD groups (P<0.0001). In the alpha1-COPD group, 24/25 patients had lower lobe emphysema. In the usual COPD group, 4 patients had upper lope predominance, 5 patients had heterogeneous distributions, and 16 patients had lower lobe predominance. CONCLUSIONS: The majority of patients with smoking related emphysema have a homogeneous distribution and lower lobe predominance although not as noticeable as in alpha1-antitrypsin deficiency. An objective and quantitative method for determining the distribution of emphysema should be applied when selecting candidates for LVRS. PMID- 15882942 TI - Wood-polyethylene composites using ethylene-vinyl alcohol copolymer as adhesion promoter. AB - Saw dust-reinforced linear low density polyethylene (LLDPE) (1:1) composites were prepared by using ethylene-vinyl alcohol copolymer (EVAL) as adhesion promoter to improve mechanical strength. To evaluate the optimum vinylalcohol (VA) content in EVAL, various EVAL samples containing different contents of VA were used. The tensile properties of saw dust-LLDPE composites were improved by using EVAL as adhesion promoter in place of ethylene-vinyl acetate copolymer (EVAc). The saw dust-LLDPE composites prepared with EVAL containing 15 mol% VA showed the maximum yield stress and modulus. The tensile stress increased with addition of EVAL up to 3wt% on the wood filler, and then leveled off in the range of 3-10 wt%. However, the elongation was decreased with increasing VA content. Hydrogen bonding interaction between saw dust and EVAL was detected by FT-IR spectra. When EVAL consisting with 15 mol% VA was used, good adhesion between saw dust and LLDPE matrix was confirmed by SEM fractography. PMID- 15882943 TI - Sequential batch culture studies for the decolorisation of reactive dye by Coriolus versicolor. AB - The white rot fungus Coriolus versicolor could decolorise reactive dye Remazol Brilliant Violet to almost 90%. The fungal mycelia removed color as well as COD up to 95% and 75%, respectively, in a batch reactor. Decolorising activity was observed during the repeated reuse of the fungus. It was possible to substantially increase the dye decolorising activity of the fungus by carefully selecting the operational conditions such as media composition, age of fungus and nitrogen source. The fungal pellets could be used for eight cycles during the long term operation, where medium and dye was replenished at the end of each cycle and the fungus was recycled. Presence of a nitrogen source and nutrient content of media played an important role in sustaining the decolorisation activity of the fungus. The form of nitrogen source (e.g. peptone vs. urea) was also important to maintain the decolorising activity with peptone showing better decolorisation. PMID- 15882944 TI - Chemical regulation of abscisic acid catabolism in plants by cytochrome P450 inhibitors. AB - Plant hormone abscisic acid (ABA) is an important factor for conferring drought stress resistance on plants. Therefore, small molecules that regulate ABA levels in plants can be useful both for investigating functions of ABA and for developing new plant growth regulators. Abscisic acid (ABA) catabolism in plants is primarily regulated by ABA 8'-hydroxylase, which is a cytochrome P450 (P450). We tested known P450 inhibitors containing a triazole group and found that uniconazole-P inhibited ABA catabolism in cultured tobacco Bright Yellow-2 cells. In a structure-activity study of uniconazole, we found a more effective ABA catabolic inhibitor (diniconazole) than uniconazole-P. Diniconazole, a fungicide, acted as a potent competitive inhibitor of recombinant Arabidopsis ABA 8' hydroxylase, CYP707A3, in an in vitro assay. Diniconazole-treated plants retained a higher ABA content and higher transcription levels of ABA response genes during rehydration than did untreated plants and were more drought stress tolerant than untreated plants. These results strongly suggest that ABA catabolic inhibitors that target ABA 8'-hydroxylase can regulate the ABA content of plants and conferred drought stress resistance on plants. The optical resolution of diniconazole revealed that the S-form isomer, which is a weak fungicidal isomer, was more active as an ABA catabolic inhibitor than was the R-form isomer. PMID- 15882945 TI - Expression of GAD65 and GAD67 immunoreactivity in MPTP-treated monkeys with or without L-DOPA administration. AB - This study investigated the consequences of levodopa treatment on the expression of the 65- and 67-kDa isoforms of glutamate decarboxylase (GAD65 and GAD67) immunoreactivity in the basal ganglia and cortex of monkeys rendered Parkinsonian by systemic MPTP administration. All MPTP-treated monkeys showed Parkinsonian impairment and selective loss of tyrosine hydroxylase (TH) with sparing of GAD immunoreactive (-ir) fibers and terminals in basal ganglia. The distribution of GAD65- and GAD67-ir in the cortex, caudate, and putamen was not significantly different in MPTP vs. naive monkeys nor as a function of L-DOPA treatment. In comparison, the expression of GAD67- but not GAD65-ir was augmented in the globus pallidus in MPTP-treated monkeys. Quantification revealed significant increases in number of GAD67-ir neurons in the external and internal segments of the globus pallidus while no significant difference in the number of GAD65-ir neurons was observed. L-DOPA treatment did not significantly change the number of GAD65- or GAD67-ir pallidal neurons following MPTP. These results support and extend the findings that transcriptional elevation of GAD67 occurs in the globus pallidus and demonstrate that GAD65 and GAD67 are differentially altered following lesion. The finding of elevated GAD67 expression in the pallidum is consistent with alterations in inhibitory neurocircuitry playing a key role in the pathophysiology of motor disturbances in Parkinson's disease. PMID- 15882946 TI - Lipopolysaccharide and lipoteichoic acid induce different innate immune responses in bovine mammary epithelial cells. AB - The objective of the present study was to characterize the innate immune responses induced by in vitro stimulation of bovine primary mammary epithelial cells (bMEC) using gram-negative lipopolysaccharide (LPS) and gram-positive lipoteichoic acid (LTA) bacterial cell wall components. Quantitative real-time PCR (qRT-PCR) was employed to examine the mRNA expression of a panel of 22 cytokines, chemokines, beta-defensins and components of the Toll-Like Receptor signaling pathway. Stimulation of bMEC with LPS for 24h elicited a marked increase in mRNA expression for IL-1beta, IL-8, TNFalpha, CXCL6 and beta-defensin while members of the Toll-Like Receptor pathway, although present, were largely unaffected. Surprisingly, stimulation of these cells with LTA for 24 h did not significantly alter the expression of these genes. A time course of the expression of IL-1beta, IL-8, TNFalpha, CXCL6 and beta-defensin was subsequently performed. The mRNA levels of all genes increased rapidly after stimulation for 2 4 h with both LPS and LTA but only the former treatment resulted in sustained responses. In contrast, the increased gene expression for LTA stimulated cells returned to resting levels after 8-16 h with the exception of beta-defensin, which remained up-regulated. The limited and unsustained cytokine response to LTA may explain why mastitis caused by gram-positive bacteria has greater potential for chronic intra-mammary infection than gram-negative infection. It was concluded that bovine mammary epithelial cells have a strong but differential capacity to mount innate immune responses to bacterial cell wall components. PMID- 15882947 TI - Global ischemia downregulates the function of metabotropic glutamate receptor subtype 5 in hippocampal CA1 pyramidal neurons. AB - Within the hippocampus, electrophysiological and immunohistochemical studies showed that metabotropic glutamate receptor subtype 5 (mGluR5) is the major postsynaptic mGluR expressed in CA1 pyramidal neurons. To better understand the role of mGluR5 in ischemia-induced neuronal death, whole-cell patch-clamp recordings using hippocampal slices were performed to investigate the functional change of mGluR5 in CA1 pyramidal neurons following transient global ischemia. Our results indicated that 6 to 24 h after global ischemia, mGluR5-induced cationic currents and mGluR5-mediated enhancement of NMDA-evoked currents in CA1 pyramidal neurons were significantly reduced. Further TaqMan real-time quantitative RT-PCR assay showed that mGluR5 mRNA expression in hippocampal CA1 region or single CA1 pyramidal neurons was significantly downregulated following ischemic insults. The present study suggests that transient global ischemia downregulates mGluR5 function of CA1 pyramidal neurons by decreasing mGluR5 mRNA and that the resulting reduced mGluR5-mediated excitotoxicity could contribute to the survival of CA1 pyramidal neurons after ischemic insult. PMID- 15882948 TI - Cell death caused by hyper-expression of a secretory exoglucanase in Escherichia coli. AB - Induced expression of a gene fusion between the ompA leader sequence and the Cellulomonas fimi cex gene encoding a secretory exoglucanase, Exg, engineered in the Tac-cassette excretion vector was lethal to Escherichia coli. An exponentially growing culture harboring the recombinant construct suffered slow growth and 99.9% of its cells died within 60-100 min after induction. This abnormality was found to have a close correlation with the rapid increase in the relative amount of the OmpA/Exg fusion precursor (Pre-Exg) compared to its processed product (Mat-Exg). Analysis of subcellular fractions revealed the presence of Pre-Exg in the inner membrane of cultures expressing high levels but not low levels of Pre-Exg. As only Pre-Exg but not Mat-Exg was detectable in the cytoplasm, and Exg was shown by cross-linking experiments to be physically associated with the Sec proteins, it was concluded that secretion and processing of Pre-Exg took place in the SecYEG translocation machinery. The results were in line with the previous speculation that accumulation of unprocessed precursor proteins in the cytoplasmic membrane was detrimental, and supported the idea that cell death was caused by some unusual tie-up of Pre-Exg with the SecYEG translocation machinery, thus imposing an inhibitory effect on the secretion of endogenous secretory proteins. A new model, designated "Saturated Translocation," was proposed to explain the interchangeable lethal and non-lethal properties of Pre-Exg, and to address the possible scenarios that might occur in the course of cell death triggered by secretion of Pre-Exg. PMID- 15882949 TI - Expression and oxidative stress tolerance studies of glutaredoxin from cyanobacterium Synechocystis sp. PCC 6803 in Escherichia coli. AB - Glutaredoxin (Grx), which has been found widely in bacteria, plant, and mammalian cells, is an electron carrier for ribonucleotide reductase and a general glutathione-disulfide reductase of importance for redox regulation. The open reading frame designated ssr2061 from cyanobacterium Synechocystis sp. PCC 6803 was found as a homologous gene coding for Grx. The amino acid sequence deduced from ssr2061 shares high identity with that of Grxs from other organisms. In the present study, the protein of Grx2061 encoded by ssr2061 was successfully overexpressed as soluble fraction in Escherichia coli BL21 (DE3). The recombinant protein was purified to near homogenity by two steps involving immobilized metal affinity chromatography and gel filtration chromatography with a yield of 22% and a specific activity of 41.5 micromol NADPH oxidized per milligram of protein in the 2-hydroxyethyl disulfide assay. The pET-2061 transformed Escherichia coli cells showed higher Grx activity and tolerance to H(2)O(2) mediated growth inhibition compared to cells transformed with the vector alone. This suggests that overexpression of Grx from Synechocystis sp. PCC 6803 may provide protection to E. coli cells against oxidative stress mediated by H(2)O(2). PMID- 15882950 TI - Cloning and expression of Klebsiella phage K11 lysozyme gene. AB - Previously, the lysozyme gene of the Klebsiella phage K11 was partially sequenced in our lab. Using the sequence information the lysozyme gene of the Klebsiella phage K11 was amplified and cloned using the polymerase chain reaction of the pfu DNA polymerase. The nucleotide sequence of phage K11 lysozyme gene was determined. The open reading frame corresponds to a polypeptide with 151 amino acids and molecular weight of 16,932 Da. The deduced amino acid sequence of this polypeptide shows 74-75% homologies to the T7 and T3 phage lysozymes. Although the gene was efficiently expressed under the control of tac promoter in Escherichia coli XL1-blue cells at 37 degrees C, most of the K11 lysozyme produced was insoluble. When the temperature of cell growth was lowered, however, solubility of the K11 lysozyme was increased gradually. The insoluble protein expressed at 37 degrees C was solubilized in 5 M guanidine-HCl and refolded in the presence of oxido-shuffling agent (GSH/GSSG). Through the refolding process the recombinant lysozyme was solubilized and purified. The purified K11 lysozyme showed transcription inhibition of K11 RNA polymerase as well as amidase activity. These results showed that the lysozyme of bacteriophage K11 is a bifunctional protein that cuts a bond in the bacterial cell wall and selectively inhibits K11 phage RNA polymerase. Also, transcription inhibition ability of K11 lysozyme with T7 or SP6 phage RNA polymerase was measured. T7 RNA polymerase was less inhibited than K11 RNA polymerase by K11 lysozyme. But SP6 RNA polymerase was not nearly inhibited by K11 lysozyme. PMID- 15882951 TI - Increase of soluble expression in Escherichia coli cytoplasm by a protein disulfide isomerase gene fusion system. AB - Human protein disulfide isomerase (PDI) was selected as a fusion partner to construct a gene expression system to enhance the solubility of recombinant protein in Escherichia coli. DREBIII-1, a plant specific transcriptional factor, was found to mainly form inclusion bodies when expressed in either His-tagged or GST-fusion systems in E. coli. In contrast, when fused with PDI, the expressed DREBIII-1 was in a highly soluble and biologically active form. Two fusion proteins, HDP and HPD, were generated by positioning DREBIII-1 at the N-terminal and C-terminal of PDI, respectively. After purification, HDP exhibited a higher stability and showed only one band on SDS-PAGE, while HPD degraded as several bands. HDP was verified to have the biological function of PDI by isomerase activity assay; meanwhile, it also presented the DNA binding and transcriptional activation characteristic of DREBIII-1 in fluorescence quenching and yeast one hybrid experiments. The PDI fusion expression system was demonstrated to be highly efficient in generating not only soluble but functional desired proteins. PMID- 15882952 TI - High-level expression and purification of a nonmitogenic form of human acidic fibroblast growth factor in Escherichia coli. AB - To decrease the potential side effects of acidic fibroblast growth factor (aFGF) caused by its broad-spectrum mitogenic activity, a nonmitogenic form of aFGF (nhaFGF), which retained the cardio- and neuroprotective characters of the wild type aFGF, was overexpressed in Escherichia coli. The expression level of nhaFGF was up to 25% of the total cellular protein. The expressed nhaFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. The mitogenic activity of the purified nhaFGF was decreased dramatically comparable to that of the wild-type aFGF (haFGF) detected by methylthiazoletetrazolium method. The purified recombinant nhaFGF was sufficiently prepared and sufficient for the following pharmacological study. PMID- 15882953 TI - Visual masking in magnetic resonance imaging. AB - When a brief target picture is followed by another picture (mask), people often report that they are not consciously aware of the target. Thus, visual masking can be used to manipulate perceptual awareness of target pictures. To avoid interference with magnetic resonance imaging, pictures have been presented on liquid crystal device (LCD) and thin film transistor (TFT) projectors that were placed outside of the scanner room. However, we found that display devices with LCD/TFT technology exhibit poor accuracy in presenting pictures at brief durations [Wiens, S., Fransson, P., Dietrich, T., Lohmann, P., Ingvar, M., Ohman, A., 2004. Keeping it short: A comparison of methods for brief picture presentation. Psychological Science, 15, 282--285]. In this paper, we present a reliable and valid masking procedure involving two LCD/TFT projectors in combination with mechanical shutters. Because LCD/TFT projectors present pictures in steady state at longer durations (e.g., after 70 ms), picture presentation is more ecologically valid than for common cathode ray tube (CRT) monitors that present pictures in multiples of refresh cycles. Also, because picture presentation with mechanical shutters is instantaneous and reliable in terms of onset, rise time, and duration, shutters can be used to control picture durations precisely in steps of milliseconds. In this paper, we also discuss risks for confounding effects from unreliable picture presentations in masking. Our findings and arguments recommend the use of mechanical shutters in front of LCD/TFT projectors in imaging studies of visual masking. PMID- 15882954 TI - Systematic approach to cutoff frequency selection in continuous-wave electron paramagnetic resonance imaging. AB - This article describes a systematic method for determining the cutoff frequency of the low-pass window function that is used for deconvolution in two-dimensional continuous-wave electron paramagnetic resonance (EPR) imaging. An evaluation function for the criterion used to select the cutoff frequency is proposed, and is the product of the effective width of the point spread function for a localized point signal and the noise amplitude of a resultant EPR image. The present method was applied to EPR imaging for a phantom, and the result of cutoff frequency selection was compared with that based on a previously reported method for the same projection data set. The evaluation function has a global minimum point that gives the appropriate cutoff frequency. Images with reasonably good resolution and noise suppression can be obtained from projections with an automatically selected cutoff frequency based on the present method. PMID- 15882955 TI - Differential gene expression of IGF-I, IGF-II, and toll-like receptors 3 and 5 during embryogenesis in hybrid (channel x blue) and channel catfish. AB - Insulin-like growth factors-I and-II (IGF-I and IGF-II) play important roles in growth and development of mammals. Toll-like receptors (TLRs) are pattern recognition molecules that orchestrate the induction of early innate immune response by recognition of specific sequences. Evidence is growing that suggests a relationship between growth and immune function. The objective of the study was to examine changes in gene expression of IGF-I, IGF-II, TLR3, and TLR5 during embryogenesis and early larval development in hybrid (channel catfishxblue catfish) and channel catfish. Egg samples were taken pre- and post-fertilization; embryos were collected at two stages of embryogenesis, at hatch, and at swim-up. All genes were detected in unfertilized catfish eggs. Expression levels of TLR5 and IGF-I mRNA in channel catfish and expression levels of TLR3, IGF-I, and IGF II mRNA in hybrids increased over time (P<0.01). Effect of time was not significant for expression of IGF-II or TLR3 mRNA in channel catfish and for TLR5 mRNA in hybrid catfish. Results of this study suggest growth (IGF-I and IGF-II) and immune (TLR3 and TLR5) associated genes could be functional and play important roles during embryogenesis and early development of hybrid and channel catfish. PMID- 15882956 TI - Clinical usefulness of the Parkinson's disease sleep scale. AB - OBJECTIVE: To test the usefulness of the Parkinson's disease sleep scale (PDSS) in identifying sleep disorders in the clinical practice setting. METHODS: Sixty two PD patients were evaluated with the PDSS and the Epworth sleepiness scale (ESS). A cut-off of less than five for each PDSS item as an indicator of substantial sleep disturbance was chosen. If the ESS was equal to or greater than eight, patients were referred to a sleep disorder specialist and possible polysomnography. RESULTS: The mean total PDSS score was 104.7+/-21.5,which correlated with the mean Hoehn and Yahr score (1.9+/-0.9) as well as the mean ESS score (9.7+/-4.7). A significant correlation was also found between the ESS score and several items of the PDSS. CONCLUSIONS: The PDSS was useful in identifying sleep disturbances which were not previously diagnosed, such as sleep maintenance insomnia and excessive daytime sleepiness. Problems with the PDSS include ambiguities of some questions, lack of quantification and an inability to identify specific sleep disturbances such as sleep apnea. PMID- 15882957 TI - Relationship between Parkinson disease with dementia and dementia with Lewy bodies. AB - Parkinson disease (PD) and dementia with Lewy bodies (DLB) are considered Lewy body diseases (LBDs). To clarify the relation between PD with dementia (PDD) and DLB, 30 patients with LBD were divided into pathological subtypes according to the consensus guidelines for DLB. Patients with PDD showed neocortical and limbic type of LBD as well as patients with DLB. Dementia had not been noted in 2 patients with neocortical type. Our results indicate that PDD and DLB share a common pathological substrate and that the pathological subtypes of LBD show considerable overlap in clinical manifestations. PMID- 15882958 TI - Molecular diversity and function of voltage-gated (Kv) potassium channels in epithelial cells. AB - Voltage-gated K+ channels belonging to Kv1-9 subfamilies are widely expressed in excitable cells where they play an essential role in membrane hyperpolarization during an action potential and in the propagation of action potentials along the plasma membrane. Early patch clamp studies on epithelial cells revealed the presence of K+ currents with biophysical and pharmacologic properties characteristic of Kv channels expressed in excitable cells. More recently, molecular approaches including PCR and the availability of more selective antibodies directed against Kv alpha and auxiliary subunits, have demonstrated that epithelial cells from various organ systems, express a remarkable diversity Kv channel subunits. Unlike neurons and myocytes however, epithelial cells do not typically generate action potentials or exhibit dynamic changes in membrane potential necessary for activation of Kv alpha subunits. Moreover, the fact that many Kv channels expressed in epithelial cells exhibit inactivation suggest that their activities are relatively transient, making it difficult to ascribe a functional role for these channels in transepithelial electrolyte or nutrient transport. Other proposed functions have included (i) cell migration and wound healing, (ii) cell proliferation and cancer, (iii) apoptosis and (iv) O2 sensing. Certain Kv channels, particularly Kv1 and Kv2 subfamily members, have been shown to be involved in the proliferation of prostate, colon, lung and breast carcinomas. In some instances, a significant increase in Kv channel expression has been correlated with tumorogenesis suggesting the possibility of using these proteins as markers for transformation and perhaps reducing the rate of tumor growth by selectively inhibiting their functional activity. PMID- 15882959 TI - Effect of dioxane on the structure and hydration-dehydration of alpha chymotrypsin as measured by FTIR spectroscopy. AB - A new experimental approach based on FTIR spectroscopic measurements was proposed to study simultaneously the adsorption/desorption of water and organic solvent on solid enzyme and corresponding changes in the enzyme secondary structure in the water activity range from 0 to 1.0 at 25 degrees C. The effect of dioxane on the hydration/dehydration and structure of bovine pancreatic alpha-chymotrypsin (CT) was characterized by means of this approach. Dioxane sorption exhibits pronounced hysteresis. No sorbed dioxane was observed at low water activities (a(w)<0.5) during hydration. At a(w) about 0.5, a sharp increase in the amount of sorbed dioxane was observed. Dioxane sorption isotherm obtained during dehydration resembles a smooth curve. In this case, CT binds about 150 mol dioxane/mol enzyme at the lowest water activities. Three different effects of dioxane on the water binding by the initially dried CT were observed. At a(w)<0.5, water adsorption is similar in the presence and absence of dioxane. It was concluded that the presence of dioxane has little effect on the interaction between enzyme and tightly bound water at low a(w). At a(w)>0.5, dioxane increases the amount of water bound by CT during hydration. This behavior was interpreted as a dioxane assisted effect on water binding. Upon dehydration at low water activities, dioxane decreases the water content at a given a(w). This behavior suggests that the suppression in the uptake of water during dehydration may be due to a competition for water-binding sites on chymotrypsin by dioxane. Changes in the secondary structure of CT were determined from infrared spectra by analyzing the structure of amide I band. Dioxane induced a strong band at 1628 cm(-1) that was assigned to the intermolecular beta-sheet aggregation. Changes in the intensity of the 1628 cm(-1) band agree well with changes in the dioxane sorption by CT. An explanation of the dioxane effect on the CT hydration and structure was provided on the basis of hypothesis on water-assisted disruption of polar contacts in the solid enzyme. The reported results demonstrate that the hydration and structure of alpha-chymotrypsin depend markedly on how enzyme has been hydrated - whether in the presence or in the absence of organic solvent. A qualitative model was proposed to classify the effect of hydration history on the enzyme activity-a(w) profiles. PMID- 15882960 TI - Pressure-tuning FT-IR spectroscopic study on the helix-coil transition of Ala rich oligopeptide in aqueous solution. AB - We investigated the effect of pressure on the helix-coil transition of an Ala rich peptide (AK16: YGAAKAAAAKAAAAKA-NH(2)) in aqueous solution by FT-IR spectroscopy. The spectra of the amide I' region of AK16 in aqueous solution was decomposed into some component bands using a curve fitting method. The peak at around 1635 cm(-1) corresponding to the solvent exposed alpha-helix conformer increases with increasing pressures, while the peak at around 1655 cm(-1) corresponding to the random coil conformer decreases. From the pressure dependence of the band intensities, we determined the volume change from the alpha-helix to random coil conformers of AK16 to be +10.5+/-0.3 cm(3)/mol. The positive volume change is different from the negative volume change generally observed in the pressure denaturation of proteins. PMID- 15882961 TI - Regulation of tyrosinase by tetrahydropteridines--what is real? A critical reanalysis of H. Wojtasek's view. PMID- 15882962 TI - Interaction of the 106-126 prion peptide with lipid membranes and potential implication for neurotoxicity. AB - Prion diseases are fatal neurodegenerative disorders characterized by the accumulation in the brain of an abnormally misfolded, protease-resistant, and beta-sheet rich pathogenic isoform (PrP(SC)) of the cellular prion protein (PrP(C)). In the present work, we were interested to study the mode of prion protein interaction with the membrane using the 106-126 peptide and small unilamellar lipid vesicles as model. As previously demonstrated, we showed by MTS assay that PrP 106-126 induces alterations in the human neuroblastoma SH-SY5Y cell line. We demonstrated for the first time by lipid-mixing assay and by the liposome vesicle leakage test that PrP 106-126, a non-tilted peptide, induces liposome fusion thus a potential cell membrane destabilization, as supported by membrane integrity assay (LDH). By circular dichroism (CD) analysis we showed that the fusogenic property of PrP 106-126 in the presence of liposome is associated with a predominantly beta-sheet structure. These data suggest that the fusogenic property associated with a predominant beta-sheet structure exhibited by the prion peptides contributes to the neurotoxicity of these peptides by destabilizing cellular membranes. The latter might be attached at the membrane surface in a parallel orientation as shown by molecular modeling. PMID- 15882963 TI - Growth suppression of Leydig TM3 cells mediated by aryl hydrocarbon receptor. AB - Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin induces developmental toxicity in reproductive organs. To elucidate the function of AhR, we generated stable transformants of TM3 cells overexpressing wild-type aryl hydrocarbon receptor (AhR) or its mutants which carried mutations in nuclear localization signal or nuclear export signal. In the presence of 3-methylcholanthrene (MC), proliferation of the cells transfected with wild-type AhR was completely suppressed, whereas cells expressing AhR mutants proliferated in a manner equivalent to control TM3 cells, suggesting AhR-dependent growth inhibition. The suppression was associated with up-regulation of cyclin-dependent kinase inhibitor p21Cip1, which was abolished by pretreatment with actinomycin D. A p38 MAPK specific inhibitor, SB203580, blocked the increase of p21Cip1 mRNA in response to MC. Treatment with indigo, another AhR ligand, failed to increase of p21Cip1 mRNA, although up-regulation of mRNA for CYP1A1 was observed. These data suggest AhR in Leydig cells mediates growth inhibition by inducing p21Cip1. PMID- 15882964 TI - Role of Rho family GTPases in CCR1- and CCR5-induced actin reorganization in macrophages. AB - The beta-chemokines, MIP-1alpha/CCL3, MIP-1beta/CCL4, and RANTES/CCL5, play a critical role in the selective accumulation and activation of macrophages in inflamed tissues. Herein, we demonstrate that the binding of each of these beta chemokines to their cognate receptors, CCR1 and CCR5, in either macrophages or in CCR1- or CCR5-transfected CHO cells, induced actin reorganization and the formation of lamellipodia that are characteristic of the activation of the Rho family GTPase, Rac. A dominant negative mutant of Rac, but not dominant negative mutants of RhoA or Cdc42, blocked MIP-1alpha-induced lamellipodia formation. Moreover, this MIP-1alpha-induced Rac activation and consequent lamellipodia formation is Gi- and phosphoinositide-3 kinase (PI3K)-mediated. Thus, Rac activation is critical for both CCR1- and CCR5-triggered signaling cascades mediating beta-chemokine-induced reorganization of the actin cytoskeleton, a process essential for effective recruitment and activation of macrophages in inflammation. PMID- 15882965 TI - Nanogel-quantum dot hybrid nanoparticles for live cell imaging. AB - We report here a novel carrier of quantum dots (QDs) for intracellular labeling. Monodisperse hybrid nanoparticles (38 nm in diameter) of QDs were prepared by simple mixing with nanogels of cholesterol-bearing pullulan (CHP) modified with amino groups (CHPNH2). The CHPNH2-QD nanoparticles were effectively internalized into the various human cells examined. The efficiency of cellular uptake was much higher than that of a conventional carrier, cationic liposome. These hybrid nanoparticles could be a promising fluorescent probe for bioimaging. PMID- 15882966 TI - The WD-40 repeat motif of Lgl tumor suppressor proteins associated with salt tolerance and temperature sensitivity. AB - We have recently identified mammalian homologues of lethal giant larvae (Lgl) tumor suppressor gene, rat Rgl-1 and bovine Bgl-1, and demonstrated that they can complement yeast double mutants lacking Sop1 and Sop2, yeast homologues of Lgl. These gene products are capable of regulating cellular viability in restrictive salt and temperature environments. Since Lgl family members contain the WD-40 repeat motif, we investigated its cellular functions using mouse homologue Mgl-1 in the absence of Sop1 and Sop2 in yeasts by complementation. Interestingly, mutant forms of Mgl-1 at the conserved glycine at position 450 and aspartic acid at position 453 in the most conserved WD-40 repeat motif were not able to complement, indicating that these amino acids are critical for regulating salt tolerance and temperature sensitivity in yeast. These results shed light on the important regulation of cytoskeletal complex for cellular polarity within eukaryotic cells. PMID- 15882967 TI - The mammalian heterochromatin protein 1 binds diverse nuclear proteins through a common motif that targets the chromoshadow domain. AB - The HP1 proteins regulate epigenetic gene silencing by promoting and maintaining chromatin condensation. The HP1 chromodomain binds to methylated histone H3. More enigmatic is the chromoshadow domain (CSD), which mediates dimerization, transcription repression, and interaction with multiple nuclear proteins. Here we show that KAP-1, CAF-1 p150, and NIPBL carry a canonical amino acid motif, PxVxL, which binds directly to the CSD with high affinity. We also define a new class of variant PxVxL CSD-binding motifs in Sp100A, LBR, and ATRX. Both canonical and variant motifs recognize a similar surface of the CSD dimer as demonstrated by a panel of CSD mutants. These in vitro binding results were confirmed by the analysis of polypeptides found associated with nuclear HP1 complexes and we provide the first evidence of the NIPBL/delangin protein in human cells, a protein recently implicated in the developmental disorder, Cornelia de Lange syndrome. NIPBL is related to Nipped-B, a factor participating in gene activation by remote enhancers in Drosophila melanogaster. Thus, this spectrum of direct binding partners suggests an expanded role for HP1 as factor participating in promoter-enhancer communication, chromatin remodeling/assembly, and sub-nuclear compartmentalization. PMID- 15882968 TI - ADAM22 plays an important role in cell adhesion and spreading with the assistance of 14-3-3. AB - Cellular adhesion plays important roles in a variety of biological processes. The ADAM family contains disintegrin-like and metalloproteinase-like domains which potentially have cell adhesion and protease activities. Recent studies suggest that the interaction between 14-3-3zeta and ADAM22cyt can regulate cell adhesion and spreading, therefore it has a potential role in neural development and function. 14-3-3 family has seven highly conserved members that regulate various cellular functions. Using yeast two-hybrid method, we identified that ADAM22cyt bound some other 14-3-3 family members. The interaction was further confirmed by in vitro protein pull-down assay and co-immunoprecipitation. We also found that the overexpression of exogenous ADAM22 in HEK293 cells could significantly enhance cell adhesion and spreading, compared with the truncated ADAM22 lack of 14-3-3 binding motifs. These results strongly demonstrated a functional role for ADAM22/14-3-3 in cell adhesion and spreading. PMID- 15882969 TI - Promoted electron transfer of mitoxantrone binding with DNA by cytochrome c. AB - A promoted electron transfer of an antitumor drug, mitoxantrone (MTX), intercalating into DNA duplex was successfully obtained upon addition of cytochromes c (cyt. c) in NaAc-HAc buffer solution (pH 4.5). The experimental results suggested that co-existence of MTX and cyt. c in the DNA helix is an important factor for accelerated electron transfer of MTX, where the promoter, cyt. c, operated smoothly through the DNA bridge. The UV/Vis spectroscopic experiments further confirmed the interaction process. Furthermore, a possible mechanism of such reaction was also discussed in this paper. PMID- 15882970 TI - Stachybotrydial, a potent inhibitor of fucosyltransferase and sialyltransferase. AB - Elevated expression of fucosylated glycoconjugates and fucosyltransferases (Fuc Ts) is found in various tumor cells and has been correlated with aspects of tumor progression such as cell adhesion and metastasis. Thus, fucosyltransferase inhibitors are potentially useful as anti-tumor agents. In the present study, three known spirocyclic drimanes (1, 2, and 3) were isolated from the culture broth of the fungus Stachybotrys cylindrospora. Compound 1 (stachybotrydial) exhibits potent inhibitory activity against alpha1,3-fucosyltransferase (Fuc-TV) during screening, while compounds 2 and 3 show no such inhibitory activity. Kinetic analysis indicates that compound 1 is an uncompetitive inhibitor with respect to GDP-fucose and a noncompetitive inhibitor with respect to N acetyllactosamine with Ki values of 10.7 and 9.7 microM, respectively. In addition, all three compounds also possess inhibitory activity against sialyltransferase (ST) but not against beta1,4-galactosyltransferase. These observations provide novel chemical structure information that will help in the design of novel Fuc-T and ST inhibitors. PMID- 15882971 TI - An A/G polymorphism of core 2 branching enzyme gene is associated with prostate cancer. AB - The expression of core 2 beta1,6-N-acetylglucosaminyltransferase-1 (C2GnT) is associated with development and progression of malignancy. Sequence analysis showed that the codon 152 of C2GnT has a polymorphism having GTT encoding valine or ATT encoding isoleucine. By examining the polymorphism in prostate cancer and benign prostatic hyperplasia patients, we found that the C2GnT G allele was more frequently observed in the prostate cancer group (p=0.015) than the control group. Men with the GG genotype had a 3.60-fold increased risk of prostate cancer, and men with the AG genotype had a 1.58-fold increased risk of prostate cancer compared with those with the AA genotype. The G allele was found to have a gene dosage effect for prostate cancer risk. No such risk was associated for benign prostatic hyperplasia. These results demonstrate that C2GnT A/G polymorphism is associated with the susceptibility to prostate cancer in a Japanese population. PMID- 15882972 TI - LRP1B is a negative modulator of increased migration activity of intimal smooth muscle cells from rabbit aortic plaques. AB - The migration of cultured cultured smooth muscle cells (SMCs) is regulated by the time-specific expression of members of the LDL receptor family (LRs). LRP1B, a member of LRs, modulates the catabolism of PDGF beta-receptor, affecting the migration of SMCs. An involvement of PDGF beta-receptor in atherosclerosis is focused because of its abundant expression in intimal SMCs. Here, in order to know a functional significance of LRP1B in the increased migration of intimal SMCs, the functions of three groups of cultured SMCs with different origins in atherosclerotic arteries were studied. Each group of SMCs (central, marginal or medial SMCs) was isolated from explanted pieces of central or marginal area of thickened intima, or media prepared from rabbit aortic plaques. The LRP1B expression levels were significantly decreased in intimal SMCs, particularly in marginal SMCs, compared to medial SMCs. The expression levels of LRP1B in SMCs were negatively correlated with those of PDGF beta-receptor. The level of PDGF beta-receptor-mediated phosphorylation of ERK 1/2 in central SMCs was increased to 5.2-fold with the functional inhibition of LRP1B using anti-LRP1B IgY. The antibody increased the PDGF-BB-stimulated migration and invasion activities in SMCs. The increase in the PDGF beta-receptor-mediated outgrowth activity of SMCs from the explants was also inhibited by the functional inhibition of LRP1B. These results indicate that LRP1B regulated the migration activity of SMCs through the modulation of PDGF beta-receptor-mediated pathway. The regulation of LRP1B expression is possibly involved in the activated migration of intimal SMCs in the course of atherosclerosis. PMID- 15882973 TI - Quantitative analysis of topoisomerase IIalpha to rapidly evaluate cell proliferation in brain tumors. AB - Immunohistochemical cell proliferation analyses have come into wide use for evaluation of tumor malignancy. Topoisomerase IIalpha (topo IIalpha), an essential nuclear enzyme, has been known to have cell cycle coupled expression. We here show the usefulness of quantitative analysis of topo IIalpha mRNA to rapidly evaluate cell proliferation in brain tumors. A protocol to quantify topo IIalpha mRNA was developed with a real-time RT-PCR. It took only 3 h to quantify from a specimen. A total of 28 brain tumors were analyzed, and the level of topo IIalpha mRNA was significantly correlated with its immuno-staining index (p<0.0001, r=0.9077). Furthermore, it sharply detected that topo IIalpha mRNA decreased in growth-inhibited glioma cell. These results support that topo IIalpha mRNA may be a good and rapid indicator to evaluate cell proliferate potential in brain tumors. PMID- 15882974 TI - Intraepithelial lymphocytes express junctional molecules in murine small intestine. AB - Intestinal intraepithelial lymphocytes (IEL) that reside at basolateral site regulate the proliferation and differentiation of epithelial cells (EC) for providing a first line of host defense in intestine. However, it remains unknown how IEL interact and communicate with EC. Here, we show that IEL express junctional molecules like EC. We identified mRNA expression of the junctional molecules in IEL such as zonula occludens (ZO)-1, occludin and junctional adhesion molecule (JAM) (tight junction), beta-catenin and E-cadherin (adherens junction), and connexin26 (gap junction). IEL constitutively expressed occludin and E-cadherin at protein level, while other T cells in the thymus, spleen, liver, mesenteric lymph node, and Peyer's patches did not. Gammadelta IEL showed higher level of these expressions than alphabeta IEL. The expression of occludin was augmented by anti-CD3 Ab stimulation. These results suggest the possibility of a novel role of IEL concerning epithelial barrier and communication between IEL and EC. PMID- 15882975 TI - Leptin activates STAT and ERK2 pathways and induces gastric cancer cell proliferation. AB - Although leptin is known to induce proliferative response in gastric cancer cells, the mechanism(s) underlying this action remains poorly understood. Here, we provide evidence that leptin-induced gastric cancer cell proliferation involves activation of STAT and ERK2 signaling pathways. Leptin-induced STAT3 phosphorylation is independent of ERK2 activation. Leptin increases SHP2 phosphorylation and enhances binding of Grb2 to SHP2. Inhibition of SHP2 expression with siRNA but not SHP2 phosphatase activity abolished leptin-induced ERK2 activation. While JAK inhibition with AG490 significantly reduced leptin induced ERK2, STAT3 phosphorylation, and cell proliferation, SHP2 inhibition only partially reduced cancer cell proliferation. Immunostaining of gastric cancer tissues displayed local overexpression of leptin and its receptor indicating that leptin might be produced and act locally in a paracrine or autocrine manner. These findings indicate that leptin promotes cancer growth by activating multiple signaling pathways and therefore blocking its action at the receptor level could be a rational therapeutic strategy. PMID- 15882976 TI - Resveratrol upregulates heme oxygenase-1 expression via activation of NF-E2 related factor 2 in PC12 cells. AB - Resveratrol (3,4',5-trihydroxy stilbene), a phytoalexin found in the skin and seeds of grapes, has been reported to possess anti-inflammatory, anticarcinogenic, and antioxidant activities. In this work, we assessed the ability of resveratrol to upregulate heme oxygenase-1 (HO-1) gene expression via activation of NF-E2-related factor 2 (Nrf2) in cultured PC12 cells. Nrf2 is a transcription factor involved in the cellular protection against oxidative stress through antioxidant response element (ARE)-directed induction of several phase 2 detoxifying and antioxidant enzymes, such as HO-1. Here, we report that resveratrol induces HO-1 expression via the ARE-mediated transcriptional activation of Nrf2. Moreover, PC12 cells treated with resveratrol exhibited transient activation of Akt/protein kinase B and extracellular signal-regulated protein kinase 1/2 (ERK1/2). LY294002 and U0126, pharmacological inhibitors of phosphatidylinositol 3-kinase and MEK1/2 which are upstream of Akt and ERK1/2, respectively, attenuated resveratrol-induced HO-1 expression and exhibited antioxidant effects. Taken together, the above findings suggest that resveratrol augments cellular antioxidant defense capacity through induction of HO-1 via Nrf2 ARE signaling, thereby protecting PC12 cells from oxidative stress. PMID- 15882977 TI - Proteasomal degradation of Kir6.2 channel protein and its inhibition by a Na+ channel blocker aprindine. AB - ATP-sensitive K+ channels (K(ATP):SUR2A+Kir6.2) play a pivotal role in cardiac protection against ischemia and reperfusion injury. When expressed in COS cells, Kir6.2 was short-lived with a half-life time of 1.9 h. The half-life time of Kir6.2 was prolonged by proteasome inhibitors MG132, ALLN, proteasome inhibitor 1, and lactacystine, but not at all by a lysosomal inhibitor chloroquine. MG132 also increased the level of ubiquitinated Kir6.2 without affecting its localization in the endoplasmic reticulum and Golgi apparatus. In electrophysiological recordings, MG132 augmented nicorandil-activated K(ATP) currents in COS cells expressing SUR2A and Kir6.2 as well as the same currents in neonatal rat cardiomyocytes. Like MG132, a Na+ channel blocker aprindine prolonged the half-life time of Kir6.2 and augmented K(ATP). Finally, both aprindine and MG132 inhibited the 20S proteasome activity in vitro. These results suggest a novel activity of aprindine to enhance K(ATP) currents by inhibiting proteasomal degradation of Kir 6.2 channels, which may be beneficial in the setting of cardiac ischemia. PMID- 15882978 TI - Caspase recruitment domain of procaspase-2 could be a target for SUMO-1 modification through Ubc9. AB - To identify the binding proteins that regulate the function of procaspase-2, we screened for proteins using the yeast two-hybrid method and isolated human Ubc9 and SUMO-1 as the candidates. Ubc9 and SUMO-1 interacted with the caspase recruitment domain of procaspase-2 in its N-terminal. We elucidated the covalent modification of procaspase-2 by SUMO-1 in mammalian cells by immunoprecipitation followed by Western blot analysis. Procaspase-2 and SUMO-1 were co-localized by dot-like structures in the nucleus that are related to promyelocytic leukemia bodies. Interestingly, a conjugation-deficient mutant (K60R) procaspase-2 resulted in a delay of its enzyme maturation (appearance of p12 subunit) compared to that of wild-type. Thus, the modification with SUMO-1 may play a critical role in the nuclear localization and the activation (maturation) of procaspase-2. PMID- 15882979 TI - The new vertebrate CYP1C family: cloning of new subfamily members and phylogenetic analysis. AB - Two novel CYP1 genes from teleost fish constituting a new subfamily have been cloned. These paralogous sequences are designated CYP1C1 and CYP1C2. Both genes were initially obtained from untreated scup Stenotomus chrysops tissues by RT-PCR and RACE. Scup CYP1C1 and CYP1C2 code for 524 and 525 amino acids, respectively, and share 80-81% identity at the nucleotide and amino acid levels. Orthologues of CYP1C1 and CYP1C2 were identified in genome databases for other fish species, and both CYP1B1 and CYP1C1 were cloned from zebrafish (Danio rerio). Phylogenetic analysis shows that CYP1Cs and CYP1Bs constitute a sister clade to the CYP1As. Analysis of sequence domains likely to have functional significance suggests that the two CYP1Cs in scup may have catalytic functions and/or substrate specificity that differ from each other and from those of mammalian CYP1Bs or CYP1As. RT-PCR results indicate that CYP1C1 and CYP1C2 are variously expressed in several scup organs. PMID- 15882980 TI - A zinc finger protein TZF is a novel corepressor of androgen receptor. AB - Steroid hormones control the transcriptional activity of target genes mediated by intracellular nuclear receptors, and these transcriptional activities are modulated by the combination with coactivators and corepressors. We found in this study that testicular zinc finger protein (TZF) that was a nuclear protein with a zinc finger motif of the Cys2-His2 type was a novel corepressor of androgen receptor (AR). Fusion protein with green fluorescence protein GFP formed the specific foci in nuclei and TZF-dependent foci were located close to the splicing factor compartment. In addition, TZF was recruited into AR subnuclear foci after the treatment of dihydrotestosterone. Furthermore, we revealed that TZF bound to the activation function-1 (AF-1) domain (N-terminal transactivating domain) of AR protein. Transient over-expression of TZF in COS-7 cells or LNCaP human prostatic cancer cell resulted in decreased AR activity in a ligand-dependent fashion. Moreover, a transcriptional corepressor N-CoR additively decreased the transcriptional activity of AR with TZF. These findings suggest that TZF might be a novel corepressor of AR. PMID- 15882981 TI - Nuclear localization signal of ING4 plays a key role in its binding to p53. AB - ING4, a novel member of ING family, is recently reported to interact with tumor suppressor p53 and negatively regulate the cell growth with significant G2/M arrest of cell cycle in HepG2 cells through upregulation of p53-inducible gene p21. However, which region of ING4 could have contributed to the binding to p53 remains largely unclear. Herein, the GST-pulldown experiments revealed that the middle region of ING4, a potential bipartite nuclear localization signal (NLS), could be involved in the binding to p53. Furthermore, the interaction of ING4 to p53 was abrogated in vitro and in vivo when certain mutations or the entire deletion of the NLS domain occurred. More interestingly, the mutations of the NLS domain could alter the ING4 nuclear localization, disrupt the interaction of ING4 with p53, and even, deregulate the p53-inducible gene p21 in MCF-7 cells. All data indicated that the NLS domain of ING4 is essential for the binding of ING4 to p53 and the function of ING4 associated with p53. PMID- 15882982 TI - Efficient gene silencing and cell differentiation using siRNA in mouse and monkey ES cells. AB - Small interfering RNA (siRNA) has been widely used for suppressing gene expression in various organisms. Here, we describe efficient methods to suppress target genes (EGFP or Oct4) using siRNA in mouse and monkey ES cells, and differentiation. In mouse ES cells, FACS analysis revealed that EGFP expression was suppressed in 97% of transfected cells at 48 h after transfection. In addition, cells expressed Hand1 and Cdx2, which are the marker genes of trophoblast lineage by the transient suppression of Oct4. In the case of monkey ES cells, highly efficient suppression was achieved in 98% of cells at 96 h post transfection using the Sendai virus (hemagglutinating virus of Japan, HVJ) envelope as a carrier of siRNA. These efficient transfection methods using synthetic siRNA should contribute to evaluate specific gene function in ES cells and can be used to differentiate ES cells into desired cell lineages. PMID- 15882983 TI - Signaling through CD40 ligand decreases CD80 expression on murine Langerhans cells and enhances IL-12 p40 production. AB - We previously showed that murine Langerhans cells (LC) express CD40 ligand (CD40L). In this study, we further investigated the function of CD40L on LC using agonistic antibodies and CD40L knockout (KO) mice. Signaling through CD40L decreased CD80 expression on LC 48 h after stimulation and the decrease was more remarkable in the presence of interferon-gamma (IFN-gamma). Signaling through CD40 enhanced the production of IL-12 p40 from LC, and simultaneous signaling through CD40L slightly augmented this effect. Addition of IFN-gamma further enhanced IL-12 p40 production. LC from CD40L KO mice expressed similar levels of surface molecules such as CD40, CD80, CD86, and MHC class II, compared with those from wild-type mice. However, they produced less amount of IL-12 p40 during 48 h after purification. These results suggest that signaling through CD40L on LC is important in regulating IL-12 production, which is critical for Th1 type immune responses. PMID- 15882984 TI - Purification of infectious adenovirus in two hours by ultracentrifugation and tangential flow filtration. AB - Adenoviruses are excellent vectors for gene transfer and are used extensively for high-level expression of the products of transgenes in living cells. The development of simple and rapid methods for the purification of stable infectious recombinant adenoviruses (rAds) remains a challenge. We report here a method for the purification of infectious adenovirus type 5 (Ad5) that involves ultracentrifugation on a cesium chloride gradient at 604,000g for 15 min at 4 degrees C and tangential flow filtration. The entire procedure requires less than two hours and infectious Ad5 can be recovered at levels higher than 64% of the number of plaque-forming units (pfu) in the initial crude preparation of viruses. We have obtained titers of infectious purified Ad5 of 1.35x10(10) pfu/ml and a ratio of particle titer to infectious titer of seven. The method described here allows the rapid purification of rAds for studies of gene function in vivo and in vitro, as well as the rapid purification of Ad5. PMID- 15882985 TI - Stabilization of integrin-linked kinase by binding to Hsp90. AB - Integrin-linked kinase (ILK) is a serine/threonine kinase that interacts with the cytoplasmic domain of beta-integrins and growth factor receptors in response to extracellular signals. It is a key molecule in cell adhesion, proliferation, and cell survival. We found that treating cells with specific inhibitors of the heat shock protein 90 (Hsp90) caused rapid cell detachment. Screening the responsible proteins revealed a decreased amount of ILK in Hsp90 inhibitor-treated cells. ILK was identified as a new Hsp90 client protein because it formed a complex with Hsp90 and Cdc37, and binding was suppressed by Hsp90 inhibitors. Experiments with a series of ILK-deletion mutants revealed that the amino acid residues 377-406 were required for Hsp90 binding. Dissociation of ILK from Hsp90 shortened its half-life by promoting proteasome-dependent degradation. These results indicate that Hsp90 plays an important role in the stability of ILK in cells. PMID- 15882986 TI - Identification and characterization of a selenium-dependent glutathione peroxidase in Setaria cervi. AB - Setaria cervi a bovine filarial parasite secretes selenium glutathione peroxidase during in vitro cultivation. A significant amount of enzyme activity was detected in the somatic extract of different developmental stages of the parasite. Among different stages, microfilariae showed a higher level of selenium glutathione peroxidase activity followed by males then females. However, when the activity was compared in excretory secretory products of these stages males showed higher activity than microfilariae and female worms. The enzyme was purified from female somatic extract using a combination of glutathione agarose and gel filtration chromatography, which migrated as a single band of molecular mass approximately = 20 kDa. Selenium content of purified enzyme was estimated by atomic absorption spectroscopy and found to be 3.5 ng selenium/microg of protein. Further, inhibition of enzyme activity by potassium cyanide suggested the presence of selenium at the active site of enzyme. This is the first report of identification of selenium glutathione peroxidase from any filarial parasite. PMID- 15882987 TI - Enhanced apoptotic action of trichosanthin in HIV-1 infected cells. AB - Trichosanthin (TCS) is a type 1 ribosome-inactivating protein (RIP) effective against HIV-1 replication. The mechanism is not clear. Present results suggested that the antiviral action may be partly mediated through enhanced apoptosis on infected cells. TCS induced apoptosis in normal H9 cells and this action was more potent in those infected with HIV-1. In flow cytometry study, TCS induced larger population of apoptotic H9 cells chronically infected with HIV-1 in a dose dependent manner. At TCS concentration of 25 microg/ml, 8.4% of normal H9 cells were found to be apoptotic whereas the same concentration induced 24.5% in HIV-1 chronically infected cells. Such difference was not found in the control experiments without TCS treatment. Two other studies supported this action. Cytotoxic study showed that cell viability was always lower in HIV-1 infected cells after TCS treatment, and DNA fragmentation study confirmed more laddering in infected cells. The mechanism of TCS induced apoptosis in normal or infected H9 cells is not clear. Results in this study demonstrated that TCS is more effective in inducing apoptosis in HIV-1 infected cells. This may explain in part the antiviral action of TCS. PMID- 15882988 TI - Three-dimensional image analysis of plugging at the septal pore by Woronin body during hypotonic shock inducing hyphal tip bursting in the filamentous fungus Aspergillus oryzae. AB - We observed that the filamentous fungus, Aspergillus oryzae, grown on agar media burst out cytoplasmic constituents from the hyphal tip soon after flooding with water. Woronin body is a specialized organelle known to plug the septal pore adjacent to the lysed compartment to prevent extensive loss of cytoplasm. A. oryzae Aohex1 gene homologous to Neurospora crassa HEX1 gene encoding a major protein in Woronin body was expressed as a fusion with DsRed2, resulting in visualization of Woronin body. Confocal microscopy and three-dimensional reconstruction of images visualized the septal pore as a dark region surrounded by green fluorescence of EGFP-fused secretory protein, RNase T1, on the septum. Dual fluorescent labeling revealed the plugging of the septal pores adjacent to the lysed apical compartments by Woronin bodies during hypotonic shock. Disruption of Aohex1 gene caused disappearance of Woronin bodies and the defect to prevent extensive loss of cytoplasm during hypotonic shock. PMID- 15882989 TI - MEKK1-induced apoptosis is mediated by Smac/Diablo release from the mitochondria. AB - During apoptotic stimulation, the serine threonine kinase, MEKK1, is cleaved into an activated 91 kDa kinase fragment. This cleavage is mediated by caspase 3 and leads to further caspase 3 activation and apoptosis. Forced expression of the 91 kDa kinase fragment induces apoptosis through changes in membrane potential of the mitochondria mediated by permeability transition pore opening. MEKK1 activation, however, fails to release cytochrome c from the mitochondria. Herein, we determined that overexpression of MEKK1 causes mitochondrial Smac/Diablo release correlating with MEKK1-induced apoptosis. Furthermore, using siRNA that lowers Smac/Diablo expression, MEKK1-induced apoptosis was significantly reduced. Mouse embryonic fibroblast cells lacking MEKK1 expression are also resistant to etoposide-induced mitochondrial Smac/Diablo release. In contrast, etoposide induced mitochondrial cytochrome c release was not inhibited. MEKK1 also activates the MAP kinase JNK, but MEKK1-induced mitochondrial Smac/Diablo release and apoptosis are independent of MEKK1 mediated JNK activation. Taken together, release of Smac/Diablo from the mitochondria plays a role in MEKK1-induced apoptosis. PMID- 15882990 TI - Molecular cloning and characterization of the murine gnathodiaphyseal dysplasia gene GDD1. AB - Mutations in the GDD1 gene cause gnathodiaphyseal dysplasia, a rare human skeletal syndrome with autosomal dominant inheritance. The biochemical function(s) of GDD1 protein and the molecular pathophysiology of GDD1 mutations leading to GDD have not yet been elucidated. In this study, we characterized the complete cDNA sequence and genomic organization of the mouse GDD1 gene. Analysis of GDD1 mRNA revealed a complex alternative splicing pattern, involving five exons of the GDD1 gene. GDD1 isoforms lacking conserved amino acids at the N terminus cytoplasmic tails, and with changes in transmembrane topology, are presumably associated with changes in protein functions and subcellular localizations of GDD1. We found GDD1 expression to be up-regulated during the course of myogenic differentiation in the murine pluripotent mesenchymal precursor cell line C2C12, whereas its expression was diminished during osteoblastic differentiation. These observations suggest diverse cellular roles of GDD1 protein. PMID- 15882991 TI - Structural insight into SoxC and SoxD interaction and their role in electron transport process in the novel global sulfur cycle in Paracoccus pantotrophus. AB - Microbial oxidation of reduced inorganic sulfur compounds mainly sulfur anions in the environment is one of the major reactions of the global sulfur cycle mediated by phylogenetically diverse prokaryotes. The sulfur oxidizing gene cluster (sox) of alpha-Proteobacteria comprises of at least 16 genes, which form two transcriptional units, viz., soxSRT and soxVWXYZABCDEFGH. Sequence analysis reveals that soxD gene product (SoxD) belongs to the di-heme cytochrome c family of electron transport proteins whereas soxC gene product (SoxC) is a sulfur dehydrogenase. We employed homology modeling to construct the three-dimensional structures of the SoxC and SoxD from Paracoccus pantotrophus. SoxD protein is known to interact with SoxC. With the help of docking studies we have identified the residues involved in the interaction of SoxC and SoxD. The putative active site geometries of these two proteins as well as the structural basis of the involvements of these proteins in electron transport process during the oxidation of sulfur anions are also investigated. PMID- 15882992 TI - Nucleotide binding to CARD12 and its role in CARD12-mediated caspase-1 activation. AB - CARD12 (Ipaf/Clan) is an important regulator of caspase-1 activation. It belongs to the family of the nucleotide-binding site and leucine-rich repeat (NBS-LRR) proteins. The NBS domain of the NBS-LRR proteins contains putative ATP/GTPase specific P-loop and Mg2+-binding site motifs. However, the nucleotide-binding properties and the function of the NBS domain are unknown. We developed a nucleotide-binding assay and investigated nucleotide binding to CARD12. We find that the NBS domain of CARD12 contains a nucleotide-binding pocket with specificity for ATP/dATP. A point mutation in the P-loop (K175R) of the NBS domain abolishes ATP/dATP binding. We further demonstrate that the nucleotide binding site is required for CARD12-mediated caspase-1 activation. CARD12 self association and association with procaspase-1 in transfected cells were markedly decreased by the P-loop mutation K175R. Furthermore, the P-loop mutation greatly reduced caspase-1 activation-dependent proIL-1beta processing. Thus, CARD12 function is dependent on the nucleotide-binding site. Our data provide insights into the molecular mechanisms of CARD12-mediated caspase-1 activation. PMID- 15882993 TI - Prolactin and growth hormone regulate adiponectin secretion and receptor expression in adipose tissue. AB - Adiponectin is a hormone secreted from adipose tissue, and serum levels are decreased with obesity and insulin resistance. Because prolactin (PRL) and growth hormone (GH) can affect insulin sensitivity, we investigated the effects of these hormones on the regulation of adiponectin in human adipose tissue in vitro and in rodents in vivo. Adiponectin secretion was significantly suppressed by PRL and GH in in vitro cultured human adipose tissue. Furthermore, PRL increased adiponectin receptor 1 (AdipoR1) mRNA expression and GH decreased AdipoR2 expression in the cultured human adipose tissue. In transgenic mice expressing GH, and female mice expressing PRL, serum levels of adiponectin were decreased. In contrast, GH receptor deficient mice had elevated adiponectin levels, while PRL receptor deficient mice were unaffected. In conclusion, we demonstrate gene expression of AdipoR1 and AdipoR2 in human adipose tissue for the first time, and show that these are differentially regulated by PRL and GH. Both PRL and GH reduced adiponectin secretion in human adipose tissue in vitro and in mice in vivo. Decreased serum adiponectin levels have been associated with insulin resistance, and our data in human tissue and in transgenic mice suggest a role for adiponectin in PRL and GH induced insulin resistance. PMID- 15882994 TI - Enzymatic and structural characterization of type II isopentenyl diphosphate isomerase from hyperthermophilic archaeon Thermococcus kodakaraensis. AB - Enzymatic and thermodynamic characteristics of type II isopentenyl diphosphate (IPP):dimethylallyl diphosphate (DMAPP) isomerase (Tk-IDI) from Thermococcus kodakaraensis, which catalyzes the interconversion of IPP and DMAPP, were examined. FMN was tightly bound to Tk-IDI, and the enzyme required NADPH and Mg2+ for the isomerization in both directions. The melting temperature (Tm), the change of enthalpy (deltaH(m)), and the heat capacity change (deltaC(p)) of Tk IDI were 88.0 degrees C, 444 kJ mol(-1), and 13.2 kJ mol(-1) K(-1), respectively, indicating that Tk-IDI is fairly thermostable. Kinetic parameters dramatically changed when the temperature crossed 80 degrees C even though its native overall structure was stably maintained up to 90 degrees C, suggesting that local conformational change would occur around 80 degrees C. This speculation was supported by the result of the circular dichroism analysis that showed the shift of the alpha-helical content occurred at 80 degrees C. PMID- 15882995 TI - Altered mechanism of the alkanesulfonate FMN reductase with the monooxygenase enzyme. AB - The two-component alkanesulfonate monooxygenase system from Escherichia coli is comprised of an FMN reductase (SsuE) and a monooxygenase enzyme (SsuD) that together catalyze the oxidation of alkanesulfonate to the corresponding aldehyde and sulfite products. To determine the effects of protein interactions on catalysis, the steady-state kinetic parameters for SsuE were determined in single enzyme assays and in the presence of the monooxygenase enzyme and alkanesulfonate substrate. In single-enzyme kinetic assays, SsuE followed an ordered sequential mechanism, with NADPH as the first substrate to bind and NADP+ as the last product to dissociate. However, in the presence of SsuD and octanesulfonate the kinetic mechanism of SsuE is altered to a rapid equilibrium ordered mechanism, and the Km value for FMN is increased 10-fold. These results suggest that both the SsuD enzyme and alkanesulfonate substrate are required to ensure that the FMN reductase reaction proceeds to form the ternary complex with the subsequent generation of reduced flavin transfer. PMID- 15882996 TI - XBP1 activates the transcription of its target genes via an ACGT core sequence under ER stress. AB - In mammals, the transmembrane protein kinase/endoribonuclease IRE1 is activated by endoplasmic reticulum stress and subsequently processes XBP1 mRNA to generate an active form of XBP1 protein. The spliced form of XBP1 protein acts as a transcription factor and induces the expression of ER-resident molecular chaperones. However, the mechanism for how XBP1 promotes the transcription of its target genes as well as the cis-acting elements for XBP1 during ER stress has been unclear. Recently, it was demonstrated that the expression of MDG1/ERdj4, a member of the DnaJ family, is regulated by the IRE1-XBP1 pathway. In the present report, we investigated the regulatory mechanisms of MDG1/ERdj4 gene expression by XBP1. We identified a cis-acting element in the MDG1/ERdj4 promoter region, to which XBP1 specifically binds in response to ER stress. Our results reveal a target sequence for the IRE1-XBP1 pathway under ER stress conditions. PMID- 15882997 TI - Functional redundancy of the Notch gene family during mouse embryogenesis: analysis of Notch gene expression in Notch3-deficient mice. AB - The Notch3 gene, a member of the Notch gene family, is expressed in a wide variety of tissues during development. We generated and analyzed Notch3-deficient mice to assess the in vivo role of the Notch3 gene. Consistent with previous observation of Krebs et al. [Characterization of Notch3-deficient mice: normal embryonic development and absence of genetic interactions with a Notch1 mutation, Genesis 37 (3) (2003) 139-143], the Notch3-/- mice were viable, fertile, and developed normally despite abundant expression of Notch3 in various embryonic tissues. We examined the details of Notch1, 2, and 4 expressions in the Notch3-/- embryos compared with those in wild-type embryos. As a result, we found that a deficiency in Notch3 did not affect the expression of Notch1, 2, and 4, and that either Notch1 or Notch2, or sometimes both, was always expressed in all Notch3 expressing tissues examined. These results support the idea that other Notch genes functionally compensate for Notch3 during embryonic development. We also surveyed the adult tissues of Notch3-/- mice and found significantly fewer thymocytes in 10-week-old mice. Therefore, the thymus might be a target tissue affected by Notch3 deficiency. PMID- 15882998 TI - Drosophila U6 promoter-driven short hairpin RNAs effectively induce RNA interference in Schneider 2 cells. AB - The effect of RNA interference (RNAi) is generally more potent in Drosophila Schneider 2 (S2) cells than in mammalian cells. In mammalian cells, PolIII promoter-based DNA vectors can be used to express small interfering RNA (siRNA) or short hairpin RNA (shRNA); however, this has not been demonstrated in cultured Drosophila cells. Here we show that shRNAs transcribed from the Drosophila U6 promoter can efficiently trigger gene silencing in S2 cells. By targeting firefly luciferase mRNA, we assessed the efficacy of the shRNAs and examined the structural requirements for highly effective shRNAs. The silencing effect was dependent on the length of the stem region and the sequence of the loop region. Furthermore, we demonstrate that the expression of the endogenous cyclin E protein can be repressed by the U6 promoter-driven shRNAs. Drosophila U6 promoter based shRNA expression systems may permit stable gene silencing in S2 cells. PMID- 15882999 TI - Mutations in the GTP-binding and synergy loop domains of Mycobacterium tuberculosis ftsZ compromise its function in vitro and in vivo. AB - The Mycobacterium tuberculosis FtsZ (FtsZ(TB)), unlike other eubacterial FtsZ proteins, shows slow GTP-dependent polymerization and weak GTP hydrolysis activities [E.L. White, L.J. Ross, R.C. Reynolds, L.E. Seitz, G.D. Moore, D.W. Borhani, Slow polymerization of Mycobacterium tuberculosis FtsZ, J. Bacteriol. 182 (2000) 4028-4034]. In an attempt to understand the biological significance of these findings, we created mutations in the GTP-binding (FtsZ(G103S)) and GTP hydrolysis (FtsZ(D210G)) domains of FtsZ and characterized the activities of the mutant proteins in vitro and in vivo. We show that FtsZ(G103S) is defective for binding to GTP and polymerization activities, and exhibited reduced GTPase activity whereas FtsZ(D210G) protein is proficient in binding to GTP, showing reduced polymerization activity but did not show any measurable GTPase activity. Visualization of FtsZ-GFP structures in ftsZ merodiploid strains by fluorescent microscopy revealed that FtsZ(D210G) is proficient in associating with Z-ring structures whereas FtsZ(G103S) is not. Finally, we show that Mycobacterium smegmatis ftsZ mutant strains producing corresponding mutant FtsZ proteins are non-viable indicating that mutant FtsZ proteins cannot function as the sole source for FtsZ, a result distinctly different from that reported for Escherichia coli. Together, our results indicate that optimal GTPase and polymerization activities of FtsZ are required to sustain cell division in mycobacteria and that the same conserved mutations in different bacterial species have distinct phenotypes. PMID- 15883000 TI - Identifying and characterising the Plasmodium falciparum merozoite surface protein 10 Plasmodium vivax homologue. AB - Plasmodium vivax malaria is one of the most prevalent parasitic diseases in Asia and Latin-America. The difficulty of maintaining this parasite culture in vitro has hampered identifying and characterising proteins implied in merozoite invasion of red blood cells. We have been able to identify an open reading frame in P. vivax encoding the Plasmodium falciparum merozoite surface protein 10 homologous protein using the partial sequences from this parasite's genome reported during 2004. This new protein contains 479 amino-acids, two epidermal growth factor-like domains, hydrophobic regions at the N- and C-termini, being compatible with a signal peptide and a glycosylphosphatidylinositol anchor site, respectively. The protein is expressed during the parasite's asexual stage and is recognised by polyclonal sera in parasite lysate using Western blot. P. vivax infected patients' sera highly recognised recombinant protein by ELISA. PMID- 15883001 TI - Matrix-assisted cell transfer for intervertebral disc cell therapy. AB - Cell therapy seems to be a promising way to reconstitute degenerated discs. We elucidate the basic aspects of intervertebral disc (IVD) cell therapy to estimate its potential in disc regeneration. Cell transfer efficiency and survival was quantified by luciferase expression after injection of recombinant cells into healthy, nucleotomized or mechanically degenerated rabbit IVDs in vitro, in situ or in vivo. A two-component fibrin matrix was adapted to allow injection of a fluid cell suspension that quickly polymerizes in IVDs. Thirty-five to fifty percent of matrix injected cells remained in the nucleus and transition zone in contrast to a rapid loss of medium-injected cells. Nucleotomy, which reduces intradiscal pressure, was crucial to the survival of the transferred cells over 3 days and nutritional enrichment of the fibrin matrix with potent biomolecules from serum significantly enhanced cell viability. In conclusion, advanced matrix substitutes are needed for efficient transfer and improved cell survival in the low-nutrient intradiscal environment to further improve disc cell therapy. PMID- 15883002 TI - Structural characterization of the fusion core in syncytin, envelope protein of human endogenous retrovirus family W. AB - Syncytin is a captive retroviral envelope protein, possibly involved in the formation of the placental syncytiotrophoblast layer generated by trophoblast cell fusion at the maternal-fetal interface. We found that syncytin and type I viral envelope proteins shared similar structural profiling, especially in the regions of N- and C-terminal heptad repeats (NHR and CHR). We expressed the predicted regions of NHR (41aa) and CHR (34aa) in syncytin as a native single chain (named 2-helix protein) to characterize it. 2-Helix protein exists as a trimer and is highly alpha-helix, thermo-stable, and denatured by low pH. NHR and CHR could form a protease-resistant complex. The complex structure built by the molecular docking demonstrated that NHR and CHR associated in an antiparallel manner. Overall, the 2-helix protein could form a thermo-stable coiled coil trimer. The fusion core structure of syncytin was first demonstrated in endogenous retrovirus. These results support the explanation how syncytin mediates cytotrophoblast cell fusion involved in placental morphogenesis. PMID- 15883003 TI - Functional analysis of a novel gene, DD3-3, from Dictyostelium discoideum. AB - A novel gene, DD3-3, from Dictyostelium discoideum has been isolated by an mRNA differential display between a wild-type strain AX2 and a mutant HG794 which is defective in O-glycosylation. Functional analysis of the novel gene, DD3-3, was conducted by preparing a knockout mutant, DD3-3KO, and a GST:DD3-3 fusion protein. The mutant DD3-3KO cells were allowed to develop about 1.5 h earlier than the wild-type strain AX2 cells. Northern blotting analysis of the knockout mutant cells showed a remarkable downregulation of Reg A, cAMP-dependent phosphodiesterase, and overexpression of protein tyrosine kinase (PTK) during early development and its shutdown during late development. The relationship between O-glycosylation and phosphorylation involving Reg A gene is discussed. PMID- 15883004 TI - Crystal structure of human B-type phosphoglycerate mutase bound with citrate. AB - The B-type cofactor-dependent phosphoglycerate mutase (dPGM-B) catalyzes the interconversion of 2-phosphoglycerate and 3-phosphoglycerate in glycolysis and gluconeogenesis pathways using 2,3-bisphosphoglycerate as the cofactor. The crystal structures of human dPGM-B bound with citrate were determined in two crystal forms. These structures reveal a dimerization mode conserved in both of dPGM and BPGM (bisphosphoglycerate mutase), based on which a dPGM/BPGM heterodimer structure is proposed. Structural comparison supports that the conformational changes of residues 13-21 and 98-117 determine PGM/BPGM activity differences. The citrate-binding mode suggests a substrate-binding model, consistent with the structure of Escherichia coli dPGM/vanadate complex. A chloride ion was found in the center of the dimer, providing explanation for the contribution of chloride ion to dPGM activities. Based on the structural information, the possible reasons for the deficient human dPGM mutations found in some patients are also discussed. PMID- 15883005 TI - L7/Pcp-2-specific expression of Cre recombinase using knock-in approach. AB - We report a knock-in mouse expressing Cre recombinase from the translational initiation site (ATG) of the endogenous L7/Pcp-2 gene. The resulting Cre expression matches the pattern of L7/Pcp-2 expression that is restricted to cerebellar Purkinje cells and retinal cells. Moreover, the Cre mouse showed no significant behavioral abnormality. Thus, our novel Cre mouse can be used for generation of Purkinje cells and retinal cell-specific gene expression and/or knockout in mouse using the Cre/loxP system. PMID- 15883006 TI - Inhibition of beta-catenin-mediated transactivation by flavanone in AGS gastric cancer cells. AB - Recently, data which prove that Wnt pathway activation may be an early event in multistep carcinogenesis in the stomach have been accumulating. We examined the effect of flavanone against beta-catenin/Tcf signaling in AGS gastric cancer cells. Reporter gene assay showed that flavanone inhibited beta-catenin/Tcf signaling efficiently. In addition, the inhibition of beta-catenin/Tcf signaling by flavanone in HEK293 cells transiently transfected with constitutively mutant beta-catenin gene, whose product is not phosphorylated by GSK3beta, indicates that its inhibitory mechanism was related to beta-catenin itself or downstream components. To investigate the precise inhibitory mechanism, we performed immunofluorescence, Western blot, and EMSA. As a result, our data revealed that there is no change of beta-catenin distribution and of nuclear beta-catenin levels through flavanone. In addition, the binding of Tcf complexes to DNA is not influenced by flavanone. The beta-catenin/Tcf transcriptional target gene cyclinD1 was downregulated by flavanone. These data suggest that flavanone inhibits the transcription of beta-catenin/Tcf responsive genes, by modulating Tcf activity without disrupting beta-catenin/Tcf complex formation. PMID- 15883007 TI - Cloning and characterization of sea bream Na+-K+-ATPase alpha and beta subunit genes: in vitro effects of hormones on transcriptional and translational expression. AB - The full length genes encoding the catalytic alpha and glycosylated beta subunits of the sodium pump (Na+-K+-ATPase) were cloned and characterized from silver sea bream gill. Using in vitro preparations of gill tissue it was found that growth hormone (10 and 100 ng/ml) caused an increase in subunit transcription, translation, and Na+-K+-ATPase enzyme activity. Similarly, insulin-like growth factor 1 (10 and 100 ng/ml) also caused an increase in Na+-K+-ATPase subunit amounts and enzyme activity. Cortisol (10 and 100 ng/ml) increased alpha subunit transcript and protein but did not modulate beta subunit expression or enzyme activity. Ovine prolactin did not cause any changes in Na+-K+-ATPase subunit transcription, translation or enzyme activity. This study is the first to describe how both Na+-K+-ATPase alpha and beta subunits are modulated at transcriptional and translational levels in fish osmoregulatory tissue upon exposure to hormones. PMID- 15883008 TI - Constitutive activity of endogenous receptors by inducible Gq overexpression. AB - We have developed an inducible cell line that transiently expresses Gq alpha G protein subunits in response to doxycycline. HEK293/Tet-On pBI(Gq alpha) cells worked consistently, achieving high and tightly regulated levels of Gq alpha overexpression (38-fold increase compared with non-induced cells). We investigated the possibility of using an inducible system to increase the proportion of constitutively active endogenously expressed G protein-coupled receptors (GPCRs) by overexpressing Gq alpha. Not only did we observe an increase in basal activity following doxycycline treatment, but also increased intrinsic activity of agonists such as carbachol, endothelin, lysophosphatidic acid (LPA), and bradykinin. Furthermore, carbachol and LPA potency increased following Gq alpha overexpression, as did the intrinsic activity of the partial agonist pilocarpine, observations indicative of constitutive activity. An inducible cell line, transiently expressing G proteins, can therefore be employed to induce constitutive activity of endogenously expressed GPCRs. This model system could be used to identify clinically important inverse agonists. PMID- 15883009 TI - Curcumin enhances Vinorelbine mediated apoptosis in NSCLC cells by the mitochondrial pathway. AB - Elderly lung cancer patients and those with poor performance status/co-morbid conditions are deprived of chemotherapy because of high toxicity of multidrug regimens. Human squamous cell lung carcinoma H520 cells treated with Curcumin were sensitized to the cytotoxicity caused by chemotherapeutic agent, Vinorelbine. Both caused apoptosis by increasing the protein expression of Bax and Bcl-xs while decreasing Bcl-2 and Bcl-X(L), releasing apoptogenic cytochrome c, and augmenting the activity of caspase-9 and caspase-3. Expression of Cox-2, NF-kappaB, and AP-1 was also affected. 23.7% apoptosis was induced in the H520 cells by treatment with Curcumin while Vinorelbine caused 38% apoptosis. Pre treatment with Curcumin enhanced the Vinorelbine induced apoptosis to 61.3%. The findings suggest that Curcumin has the potential to act as an adjuvant chemotherapeutic agent and enhance chemotherapeutic efficacy of Vinorelbine in H520 cells in vitro. Thus, Curcumin offers the prospect of being beneficial in the above-mentioned patient groups. PMID- 15883010 TI - Structural requirements for O2 sensing by the human tandem-P domain channel, hTREK1. AB - TREK1 is a member of the tandem-P domain K+ channel family which is expressed almost exclusively in the nervous system. It is modulated by a number of important factors including arachidonic acid and cell swelling. Since both factors are associated with brain ischemia, it has been suggested that activation of TREK1 may confer neuroprotection. However, it has been reported that the stably expressed human homologue of TREK1 is inhibited by hypoxia, calling into question its neuroprotective role in ischemia. Here, using transient transfection of HEK 293 cells with several hTREK1 mutations and whole-cell patch-clamp, we show that: hypoxic inhibition: (a) requires the C-terminal domain of the channel; (b) does not involve redox modulation of the C-terminal domain cysteine residues C365 and C399; and (c) is critically dependent on the glutamate residue at position 306. These data suggest strongly that neuroprotection is unlikely to be provided by this channel in low O2 environments and continue to cast a shadow of doubt over the precise role that TREK may have during hypoxic episodes. PMID- 15883011 TI - Cloning and expression analysis of Zmglp1, a new germin-like protein gene in maize. AB - The cDNA and genomic DNA of a green tissue-specific gene were cloned from maize (Zea mays L.) using cDNA-amplified fragment length polymorphism (cDNA-AFLP) and library screening. The deduced protein was highly similar to Hordeum vulgare germin-like protein 1 (HvGLP1), and the maize gene was therefore designated Zmglp1. Northern blot specifically detected the mRNA of Zmglp1 in young whorl leaves at the early-whorl stage. However, at the late-whorl, tassel, and silk stages, Zmglp1 transcripts were highly abundant in young whorl leaves; less abundant in mature leaves, young tassels, and cobs; and not detectable in roots, immature kernels, and stalks. RNA in situ hybridization revealed that Zmglp1 expressed only in mesophyllous, phloem, and guard cells in the young whorl leaves. Deletion analysis of the promoter in transgenic Arabidopsis resulted in the identification of several regions containing important regulatory cis elements controlling the expression levels and circadian rhythm-oscillated patterns of Zmglp1. PMID- 15883012 TI - Anti-oxidant and anti-inflammatory activities of macelignan in murine hippocampal cell line and primary culture of rat microglial cells. AB - Epidemiological studies suggest that the treatments of anti-inflammatory agents and anti-oxidants slow the progress of neurological diseases. Lignans are anti oxidants and phytoestrogens found in a variety of plants. In this study, we investigated the neuroprotective effect of macelignan on glutamate-induced neurotoxicity and reactive oxygen species (ROS) in murine hippocampal HT22 cell line. Macelignan significantly attenuated the ROS production and neurotoxicity induced by glutamate in HT22 cell. Also, the properties of macelignan as an anti inflammatory agent were investigated in microglials activation by lipopolysaccharide (LPS). It potently suppressed the expression of cyclooxygenase 2 and inducible nitric oxide synthase, that consequently resulted in the reduction of nitric oxide in LPS-treated microglial cells. It also significantly suppressed the production of pro-inflammatory cytokine tumor necrosis factor alpha and interleukin-6. These results suggest that macelignan possesses therapeutic potentials against neurodegenerative diseases with oxidative stress and neuroinflammation. PMID- 15883013 TI - Pnlip encoding pancreatic lipase is possible candidate for obesity QTL in the OLETF rat. AB - The Otsuka Long-Evans Tokushima Fatty (OLETF) rat exhibits polygenic obesity, and one of quantitative trait loci (QTLs) responsible for a susceptibility to obesity in the OLETF, Nidd6/of, has been mapped to the approximately 10-cM genomic region between D1Rat166 and D1Rat90 on chromosome 1 in (OLETF x normal) F2 intercross. In this study, we have attempted to identify the causal gene for the Nidd6/of QTL. A Nidd6/of congenic strain, constructed by introgressing the OLETF allele on the mapped Nidd6/of region in the normal F344 rat strain, confirmed the existence of the Nidd6/of as obesity QTL. The Nidd6/of region was refined to a approximately 2.3-cM genomic region between D1Rat225 and D1Rat90, using informative recombinants selected from (Nidd6/of congenic x F344) F1 x Nidd6/of congenic backcross progenies. Among 46 genes located within the approximately 2.3 cM region, pancreatic lipase gene, Pnlip, was regarded as the most prominent and physiologically relevant positional candidate for the Nidd6/of QTL. We found that Pnlip possesses an OLETF allele-specific increase of mRNA levels in the pancreas, and that the OLETF allele is longer in variable number of tandem repeat (VNTR) within the 5'-flanking region than normal alleles. We further showed that the Nidd6/of QTL completely cosegregates with Pnlip VNTR in the informative recombinants from (Nidd6/of congenic x F344) F1 x Nidd6/of congenic backcross progenies. These results suggest that Pnlip is possible candidate for the Nidd6/of QTL. PMID- 15883014 TI - Epicatechin protects endothelial cells against oxidized LDL and maintains NO synthase. AB - Intake of flavanol-rich food or beverages was previously shown to ameliorate endothelial function and to enhance bioactivity of nitric oxide with individuals at risk for cardiovascular disease. Here, we examined whether the major dietary flavanol, (-)-epicatechin, counteracts the action of oxidized LDL on endothelial cells, an action considered pivotal for endothelial dysfunction in the pathogenesis of atherosclerosis. Oxidation by myeloperoxidase plus nitrite rendered human LDL cytotoxic towards endothelial cells, more so than oxidation by Cu2+. Oxidized LDL also caused a marked loss of endothelial NO synthase protein which did not occur in the presence of a proteasome inhibitor, lactacystin. Both actions of oxidized LDL, which were not evoked by native LDL, were effectively counteracted by (-)-epicatechin. We conclude that dietary flavanols contribute to protection of the integrity of endothelial cells not only by scavenging free radicals but also by maintaining endothelial NO synthase. PMID- 15883015 TI - Partially folded intermediate state of concanavalin A retains its carbohydrate specificity. AB - A systematic investigation of the effect of polyethylene glycol (PEG) 200 and 400 on the solution conformation of concanavalin A (con A) was made using circular dichroism (CD), tryptophan fluorescence, 1-anilino-8-naphthalenesulfonic acid (ANS) binding, and size-exclusion chromatography. Far-UV CD spectra of con A at 30%(v/v) PEGs show the retention of ordered secondary structure as compared to 70%(v/v) PEGs. Near-UV CD spectra showed the retention of native-like spectral features in the presence of 30%(v/v) PEGs. Intrinsic tryptophan fluorescence studies indicate a change in the environment of tryptophan residues on the addition of PEG. ANS binding was maximum at 30%(v/v) PEGs suggesting the compact "molten-globule"-like state with enhanced exposure of hydrophobic surface area. Size-exclusion chromatography indicates an intermediate hydrodynamic size at 30%(v/v) PEGs. GdnHCl denaturation of these states was a single-step, two-state transition. To study the possible minimum structural requirement in the specific binding, the effect of PEGs on the interaction of con A with ligand was investigated by turbidity measurements. The C50 value was less in PEG 400 suggesting the more inhibitory ability of PEG 400. The C50 value of PEGs was highest for dextran followed by glycogen, ovalbumin, and ovomucoid. From percentage inhibition of con A-ligands at 30%(v/v) PEG, maximum inhibition was in ovalbumin followed by ovomucoid, glycogen, and dextran. To summarize: con A at 30%(v/v) PEGs exists as compact intermediate with molten-globule-like characteristics, viz., enhanced hydrophobic surface area, retention of compact secondary as well as tertiary structure, and a considerable degree of carbohydrate binding specificity and activity. This result has significant implications on the molten globule state during the folding pathway(s) of proteins in general and quaternary association in the legume lectin in particular, where precise topology is required for their biological activities. PMID- 15883016 TI - CXCL16 is a novel angiogenic factor for human umbilical vein endothelial cells. AB - CXCL16 is a unique chemokine with characteristics as a receptor for phosphatidylserine and oxidized low density lipoproteins in macrophages, and is involved in the accumulation of cellular cholesterol during atherosclerotic lesion development. In this study, we report a new function of CXCL16 as a novel angiogenic factor in human umbilical vein endothelial cells (HUVEC). CXCL16 stimulated proliferation and chemotaxis of HUVEC in a dose-dependent manner, reaching a maximum at 1 nM. CXCL16 also significantly induced tube formation of HUVEC on Matrigel. Further, exposure of HUVEC to CXCL16 led to a time- and dose dependent activation of mitogen-activated protein kinase (ERK1/2), which was completely inhibited by a mitogen-activated protein kinase kinase inhibitor, PD98059. Proliferation and tube formation in response to CXCL16 were also blocked by the pretreatment with PD98059, but not CXCL16-induced chemotaxis. Thus, our data indicate that CXCL16 may act as a novel angiogenic factor for HUVEC and that ERK is involved as an important signaling molecule to mediate its angiogenic effects. PMID- 15883017 TI - Possible role of calcineurin in heating-related increase of rat muscle mass. AB - The purpose of this study was to investigate the contribution of calcineurin related intracellular signal for heat-stress-associated muscle hypertrophy. Wistar strain male rats (7-week-old) were randomly divided into four groups: (1) control (CC, n=15), (2) control with the injection of cyclosporine A (CsA) (CA, n=15), (3) heat-stressed (HC, n=15), and (4) heat-stressed with the injection of CsA (HA, n=15). The heat-stress groups (HC and HA) were exposed to heat (41 degrees C for 60 min) in a controlled heat chamber without anesthesia. Soleus and extensor digitorum longus (EDL) muscles were dissected and weighed 1, 7, and 14 days after the exposure. Wet and dry weights of soleus were increased 7 days following heat exposure. The expressions of heat shock protein 72 (HSP72) and calcineurin in both muscles were also increased within 1 and 7 days following heat-stress, respectively. Administration of CsA, a specific inhibitor for calcineurin, depressed heat-stress-associated increase of muscle weight and calcineurin expression, especially in soleus. These observations suggest that a calcineurin-dependent signaling pathway may play an important role in the heat stress-related skeletal muscular hypertrophy. Application of heat-stress to skeletal muscles may be a useful tool to gain muscular mass and force generation not only in athletes, but also in patients during rehabilitation. PMID- 15883018 TI - Effects of single amino acid substitutions at the predicted coiled-coil or hydrophobic region on the self-assembly of phi29 replication protein, gp1. AB - Gp1, the product of one of the essential genes of phi29 replication, is an RNA binding protein and self-associates to form large complexes. Furthermore, gp1 suppresses the synthesis of phi29 DNA polymerase and primer protein in the post transcriptional process. In this report, we have employed seven variants with single amino acid substitutions to analyze the self-assembly of gp1. Using chemical cross-linking and sedimentation assays, amino acid substitutions within the predicted coiled-coil or hydrophobic region were shown to strongly affect the formation of large complexes, suggesting that these two regions were required for the self-assembly of gp1. The self-association of gp1 was suggested to be necessary for the efficient binding to RNA and the translational repression. PMID- 15883019 TI - Effects of T3 treatment on HSP72 and calcineurin content of functionally overloaded rat plantaris muscle. AB - The content of both heat shock protein 72 (HSP72) and calcineurin (CaN) in skeletal muscle fibers have been reported to be associated with the slow phenotype. The purpose of the present study was to determine the adaptations/contributions of HSP72 and CaN to experimental conditions producing dramatic shifts in fiber phenotype. Two weeks of functional overload (FO) of the rat plantaris by cutting the tendons of its major synergists resulted in a shift towards a slower MHC profile. Two weeks of thyroid hormone (T3) administration (150 microg/kg/day, i.p.) resulted in a shift towards a faster MHC profile in control rats and an attenuation of the shift towards a slower profile in FO rats. HSP72 and CaN-A content were 63% and 47% higher, respectively, in the plantaris of FO than age-matched control rats. These increases were significantly attenuated by T3 treatment in FO rats. CaN-B levels were approximately 50% higher in FO and FO plus T3-treated than control rats. T3 treatment alone had no effect on the levels of HSP72, CaN-A or -B in control rats. Therefore, chronic overload of a muscle results in an increase in the percentage of slow fibers/MHC and enhances the levels of HSP72 and CaN. In turn, these FO-induced adaptations are attenuated by T3 treatment. Combined, these results indicate that muscle HSP72 and CaN protein levels are modulated by mechanical stress and that their levels appear to be related to changes in fiber type/MHC composition, at least in chronically overloaded muscles. PMID- 15883020 TI - The unique glutathione reductase from Xanthomonas campestris: gene expression and enzyme characterization. AB - The glutathione reductase gene, gor, was cloned from the plant pathogen Xanthomonas campestris pv. phaseoli. Its gene expression and enzyme characteristics were found to be different from those of previously studied homologues. Northern blot hybridization, promoter-lacZ fusion, and enzyme assay experiments revealed that its expression, unlike in Escherichia coli, is OxyR independent and constitutive upon oxidative stress conditions. The deduced amino acid sequence shows a unique NADPH binding motif where the most highly conserved arginine residue, which is critical for NADPH binding, is replaced by glutamine. Interestingly, a search of the available Gor amino acid sequences from various sources, including other Xanthomonas species, revealed that this replacement is specific to the genus Xanthomonas. Recombinant Gor enzyme was purified and characterized, and was found to have a novel ability to use both, NADPH and NADH, as electron donor. A gor knockout mutant was constructed and shown to have increased expression of the organic peroxide-inducible regulator gene, ohrR. PMID- 15883021 TI - Evidence for chaperone heterocomplexes containing both Hsp90 and VCP. AB - With assistance from co-chaperone partner proteins, Hsp90 plays an essential positive role in supporting the structure and function of numerous client proteins in vivo. Hsp90's co-chaperone partnerships are believed to regulate and/or target its function. Here we describe associations between Hsp90 chaperone machinery and another chaperone, the 97-kDa valosin-containing protein VCP. Coimmunoadsorption assays indicate that VCP occurs in one or more native heterocomplexes containing Hsp90 and the Hsp90 partner proteins Cdc37, FKBP52, and p23. Functional characterizations indicate that VCP is not an Hsp90 substrate, but rather demonstrate the biochemical hallmarks of an Hsp90 co chaperone. Potential roles for a collaboration between for Hsp90 and VCP are discussed. PMID- 15883022 TI - Endurance training increases lactate transport in male Zucker fa/fa rats. AB - The purpose of this study was to investigate the effect of endurance training (10 weeks) on previously reported alterations of lactate exchange in obese Zucker fa/fa rats. We used sarcolemmal vesicles to measure lactate transport capacity in control sedentary rats, Zucker (fa/fa), and endurance trained Zucker (fa/fa) rats. Monocarboxylate transporter (MCT) 1 and 4 content was measured in sarcolemmal vesicles and skeletal muscle. Training increased citrate synthase activity in soleus and in red tibialis anterior, and improved insulin sensitivity measured by intraperitoneal glucose tolerance test. Endurance training increased lactate influx in sarcolemmal vesicles at 1 mM of external lactate concentration and increased MCT1 expression on sarcolemmal vesicles. Furthermore, muscular lactate level was significantly decreased after training in red tibialis anterior and extensor digitorum longus. This study shows that endurance training improves impairment of lactate transport capacity that is found in insulin resistance state like obesity and type 2 diabetes. PMID- 15883023 TI - Annexin1 regulates the erythroid differentiation through ERK signaling pathway. AB - K562 cells can be used as a model of erythroid differentiation on being induced by hemin. We found that the level of annexin1 gene expression was notably increased during this indicated process. To test the hypothesis that annexin1 can regulate erythropoiesis, K562 cell clones in which annexin1 was stably increased and was knocked down by RNAi were established, respectively. With analysis by hemoglobin quantification, benzidine staining, and marker gene expression profile determination, we confirmed that hemin-induced erythroid differentiation of K562 cells was modestly stimulated by overexpression of annexin1 while it was significantly blocked by knock down of annexin1. Further studies revealed that the mechanisms of annexin1 regulation of the erythroid differentiation was partially related to the increased ERK phosphorylation and expression of p21(cip/waf), since specific inhibitor of MEK blocked the function of annexin1 in erythroid differentiation. We concluded that annexin1 exerted its erythropoiesis regulating effect by ERK pathway. PMID- 15883024 TI - Vitamin D upregulates expression of ECaC1 mRNA in semicircular canal. AB - The low luminal Ca2+ concentration of mammalian endolymph in the vestibular labyrinth is required for normal balance. We found transcripts in primary cultures of semicircular canal duct (SCCD) epithelial cells from neonatal rats representing a complete transport system for transepithelial absorption of Ca2+ that is comprised of the epithelial Ca2+ channels ECaC1 (CaT2, TRPV5) and ECaC2 (CaT1, TRPV6), calbindin (calbindin-D9k, calbindin-D28k), Na+/Ca2+ exchanger (NCX1, NCX2, and NCX3), and plasma membrane Ca2+-ATPase (PMCA1, PMCA3, and PMCA4) by RT-PCR. Further, vitamin D receptor was also expressed in SCCD and it was found by quantitative RT-PCR that incubation for 24 h with 1,25-dihydroxyvitamin D3 upregulated the expression of ECaC1, calbindin-D9k, and calbindin-D28k. These observations provide evidence for the first time of an ECaC-based Ca2+ transport system in SCCD that could maintain the low Ca2+ concentration in vestibular endolymph. PMID- 15883025 TI - The antibodies directed against N-terminal heptad-repeat peptide of hRSV fusion protein and its analog-5-Helix inhibit virus infection in vitro. AB - Human respiratory syncytial virus (hRSV) membrane fusion is promoted by the formation of a trimer-of-hairpins structure that brings the amino- and carboxyl terminal regions of fusion (F) protein into close proximity. Two heptad-repeat (HR1 and HR2) regions in F protein play an important role in this process. Our previous study demonstrated that peptides derived from HR1 and HR2 regions of F protein were potent inhibitors of hRSV entry. Here we showed that HR1 peptide and its analog denoted 5-Helix which contained a central coiled-coil formed by three HR1s could induce highly potent antibody response in the immunized rabbits. Both antibodies could recognize F1 domain of the F protein and inhibited hRSV entry with the neutralizing antibody titers of 1:61 and 1:115, respectively. These suggested that 5-Helix could induce potent neutralizing antibody response and the central coiled-coil might be a highly conserved neutralization site for hRSV F protein. PMID- 15883026 TI - In vivo induced clpB1 gene of Vibrio cholerae is involved in different stress responses and affects in vivo cholera toxin production. AB - Previously in global transcription profile approach one of the cosmid clones of Vibrio cholerae containing the genes pnuC, icmF, and a fragment of clpB2 showed higher expression in V. cholerae grown inside rabbit intestine. In the present report, both the stress responsive clpB genes of V. cholerae O395 were cloned, clpB1 from chromosome I and clpB2 present in chromosome II. From the Northern blot hybridization it was observed that the level of transcription of clpB2 was very low which could be due to the weak promoter strength of clpB2 as predicted in silico. The deduced amino acid sequence showed that clpB1 possesses features typical of the ClpB ATPase family of stress response proteins. The clpB1 gene showed about three times higher expression under in vivo condition than in vitro. Increased expression of clpB1 gene was also observed at high temperature, high salt, and in the condition mimicking human intestine viz., 37 degrees C, pH 8.5, 300 mM NaCl, which is known to be the repressive condition for ToxR, the global transcriptional regulator of virulence in V. cholerae. The clpB1 insertion mutant showed increased sensitivity towards high temperature, oxidative stress, and acid pH. ClpB1 also conferred thermotolerance to V. cholerae. These effects could be reversed by complementation. Although clpB1 appeared not to be under the control of virulence regulatory cascade of V. cholerae, the CT production was reduced in clpB1 mutant when tested in vivo in an infant mice model. PMID- 15883027 TI - Molecular, biochemical, and cellular characterization of epididymal ADAMs, ADAM7 and ADAM28. AB - The mammalian epididymis is critical for sperm to acquire motility and fertilizing capacity. This maturation process involves the interaction of epididymal secretory proteins with sperm. We analyzed mouse a disintegrin and metalloprotease (ADAMs) 7 and 28 expressed specifically or predominantly in the epididymis. We found that these ADAM genes are expressed in an epididymal region specific manner and their gene expression is regulated by both androgen and testicular factors (ADAM7) or only testicular factors (ADAM28). We identified an ADAM28 transcript isoform that lacks the transmembrane domain. Protein analysis revealed that ADAM7, but not ADAM28, is transferred from the epididymis to the sperm surface and redistributed in the sperm head during acrosome reaction. These processes were shown to occur without processing of the protein. Taken together, our results indicate that the two epididymal ADAMs closely related in phylogeny are differential in various characteristics and ADAM7 has unique secretory feature and interactive relationship with sperm. PMID- 15883028 TI - Variable Meis-dependence among paralog group-1 Hox proteins. AB - Optimal function of Hox transcription factors may require Meis and Pbx cofactors. Here we test the in vivo Meis-dependence of two zebrafish paralog group-1 (PG1) Hox proteins. Misexpression of Hoxb1a induces ectopic gene expression throughout the anterior nervous system, while Hoxb1b induces ectopic expression primarily in hindbrain rhombomere 2. These activities are drastically reduced when endogenous Meis function is disrupted, demonstrating that both proteins are Meis-dependent. Upon addition of Meis3, Hoxb1b mimics the more severe Hoxb1a phenotype, indicating that Hoxb1b requires higher Meis levels than Hoxb1a. Using chimeric proteins we map this difference to the N-terminus, which contains the transcription activation domain. Lastly, we demonstrate strong genetic interactions between meis and PG1 hox genes, as well as between meis and pbx genes, in vivo. Our results are consistent with PG1 hox genes requiring pbx and meis function in vivo and reveal that different Hox proteins have distinct Meis requirements. PMID- 15883029 TI - Proteasome modulating agents induce rAAV2-mediated transgene expression in human intestinal epithelial cells. AB - Intestinal gene transfer offers promise as a therapeutic option for treatment of both intestinal and non-intestinal diseases. Recombinant adeno-associated virus serotype 2, rAAV2, based vectors have been utilized to transduce lung epithelial cells in culture and in human subjects. rAAV2 transduction of intestinal epithelial cells, however, is limited both in culture and in vivo. Proteasome inhibiting agents have recently been shown to enhance rAAV2-mediated transgene expression in airway epithelial cells. We hypothesized that similar inhibition of proteasome-related cellular processes can function to induce rAAV2 transduction of intestinal epithelial cells. Our results demonstrate that combined treatment with proteasome-modulating agents MG101 (N-acetyl-L-leucyl-L-leucyl-L-norleucine) and Doxorubicin synergistically induces rAAV2-mediated luciferase transgene expression by >400-fold in undifferentiated Caco-2 cells. In differentiated Caco 2 monolayers, treatment with MG101 and Doxorubicin induces transduction preferentially from the basolateral cell surface. In addition to Caco-2 cells, treatment with MG101 and Doxorubicin also results in enhanced rAAV2 transduction of HT-29, T84, and HCT-116 human intestinal epithelial cell lines. We conclude that MG101 and Doxorubicin mediate generic effects on intestinal epithelial cells that result in enhanced rAAV2 transduction. Use of proteasome-modulating agents to enhance viral transduction may facilitate the development of more efficient intestinal gene transfer protocols. PMID- 15883030 TI - Cloning and characterization of a novel RNA polymerase II C-terminal domain phosphatase. AB - Reversible phosphorylation of RNA polymerase (RNAP) II's largest subunit C terminal domain (CTD) is a key event during mRNA metabolism. The CTD phosphatase, FCP1, catalyzes the dephosphorylation of RNAP II and is thought to play a major role in polymerase recycling. In this study, we isolated a novel phosphatase gene by large-scale sequencing analysis of a human fetal brain cDNA library. Its cDNA is 2215 bp in length, encoding a 318-amino acid polypeptide that contains a ubiquitin-like domain and a CTD phosphatase domain. Therefore, it was termed ubiquitin-like domain containing CTD phosphatase 1 (UBLCP1). Reverse transcription PCR (RT-PCR) revealed that UBLCP1 was expressed with relatively lower levels in most adult normal tissues and higher levels in fast growing or tumor tissues. Transient transfection experiment suggested that UBLCP1 was localized in the nucleus of COS-7 cells. Significantly, UBLCP1 could dephosphorylate GST-CTD in vitro. Accordingly, UBLCP1 may play a role in the regulation of phosphorylation state of RNA polymerase II C-terminal domain. PMID- 15883031 TI - Functional co-localization of monocytic aminopeptidase N/CD13 with the Fc gamma receptors CD32 and CD64. AB - Information about the function of aminopeptidase N/CD13 on monocytes is limited. In order to gain more insight into its interaction with other proteins, we have identified molecules that co-localize with the membrane ectoenzyme at the cell surface of monocytes. Using laser scanning and electron microscopy as well as fluorescence resonance energy transfer (FRET) measured by flow cytometry we show that monocytic CD13 co-localized with the Fc gamma receptor II/CD32 after Fc receptor ligation by a CD32-specific antibody. FRET was also observed between CD13 and the Fc gamma receptor I/CD64, but not with the myeloid marker CD33 representing a member of the sialoadhesin family. Our results imply a novel functional role of CD13 and Fc gamma receptors as members of a multimeric receptor complex. Further studies have to be done to elucidate common signaling pathways of these molecules. PMID- 15883032 TI - Legionella pneumophila Philadelphia-1 tatB and tatC affect intracellular replication and biofilm formation. AB - Legionella pneumophila is a facultative intracellular human pathogen and an important cause of Legionnaires' disease, a severe form of pneumonia. Recently, we showed the presence of a putative twin-arginine translocation (Tat) pathway in L. pneumophila Philadelphia-1. This secretion pathway is used to transport completely folded proteins across the cytoplasmic membrane. The importance of the Tat pathway in L. pneumophila was investigated by constructing a tatB and a tatC mutant. Functionality of the Tat pathway was shown using a proven heterologous Tat substrate. It was shown that tatB and tatC are involved in intracellular replication in Acanthamoeba castellanii and differentiated U937 cells, and in biofilm forming ability. A putative Legionella Tat substrate was identified via 2D gel electrophoresis. PMID- 15883033 TI - AP-1 mediates beta-amyloid-induced iNOS expression in PC12 cells via the ERK2 and p38 MAPK signaling pathways. AB - Nitrosative stress with subsequent inflammatory cell death has been implicated in some neurodegenerative disorders such as Alzheimer's disease (AD). Expression of inducible nitric oxide synthase (iNOS) and production of nitric oxide (NO) have been frequently elevated in AD. In this study, we have investigated the molecular mechanisms underlying nitrosative stress induced by beta-amyloid (A beta), a neurotoxic peptide associated with senile plaques formed in the brains of patients with AD. Exposure of rat pheochromocytoma (PC12) cells to the A beta resulted in increased mRNA and protein expression of iNOS and generation of NO. NO can rapidly interact with superoxide anion, forming more reactive peroxynitrite. Treatment of PC12 cells with A beta led to increased peroxynitrite production and nitrotyrosine formation. A beta induced activation of redox sensitive transcription factor activator protein-1 (AP-1), and AP-1 antisense oligonucleotide abolished the A beta-induced iNOS expression. Moreover, A beta transiently activated extracellular signal-regulated kinase (ERK) and p38 mitogen activated protein kinase (p38 MAPK) via phosphorylation. Pharmacologic inhibition of both enzymes or dominant-negative mutation of ERK2 or p38 MAPK effectively down-regulated DNA binding as well as transcriptional activity of AP-1 and subsequent iNOS expression and NO production. The above findings suggest that A beta induces iNOS expression in PC12 cells through activation of AP-1 which is regulated by upstream kinases, such as ERK and p38 MAPK. PMID- 15883034 TI - Autocrine/paracrine stimulation of purinergic receptors in osteoblasts: contribution of vesicular ATP release. AB - Extracellular nucleotides such as ATP and UTP are released in response to mechanical stimulation in different cell systems. It is becoming increasingly evident that ATP release plays a role in autocrine and paracrine stimulation of osteoblasts. Mechanical stimulation, as shear stress, membrane stretch or hypo osmotic swelling, as well as oscillatory fluid flow, stimulates ATP release from different osteoblastic cell lines. Human osteoblast-like initial transfectant (HOBIT) cells release ATP in response to mechanical stimulation. In the present study, we show that HOBIT cells are activated by nanomolar levels of extracellular ATP, concentrations that can be detected under resting conditions and increase following hypotonic shock. Cell activation by hypotonic medium induced intracellular Ca2+ oscillations, and Egr-1 synthesis and DNA-binding activity. Quinacrine staining of living, resting cells revealed a granular fluorescence, typical of ATP-storing vesicles. Monensin prevented quinacrine staining and considerably inhibited hypotonic-induced ATP release. Finally, elevated levels of cytosolic Ca2+ activated massive ATP release and a dose dependent loss of quinacrine granules. The contribution of a vesicular mechanism for ATP release is proposed to sustain paracrine osteoblast activation. PMID- 15883035 TI - Polymorphisms in human pre-miRNAs. AB - MicroRNAs constitute a growing class of non-coding RNAs that are thought to regulate gene expression via translational repression. MicroRNAs are initially transcribed as several hundred-nucleotide pri-miRNAs and are then processed to approximately 60-nucleotide hairpin pre-miRNAs. We hypothesized that polymorphisms in both pre-miRNA and mature microRNA modify various biological processes by influencing the processing and/or target selection of microRNAs. In the present study, we sequenced 173 human pre-miRNA genome regions in 96 subjects and found 10 polymorphisms in the 10 pre-miRNA hairpin regions. Although most of these polymorphisms seem to have no effect on microRNA processing, we identified a C to A polymorphism in the mature miR-30c-2 sequence. This polymorphism may alter target selection and thus exert profound biological effects. To the best of our knowledge, this is the first report of polymorphisms in pre-miRNAs. PMID- 15883036 TI - Identification of a protein that interacts with the vanilloid receptor. AB - The vanilloid receptor (VR1 or TRPV1) is a capsaicin (CAP)-sensitive non selective cation channel. Although its channel activity is reportedly modulated through protein-protein interactions, to date very few VR1 interacting proteins have been identified. To address this issue, a yeast two-hybrid screening technique using the C-terminus of rVR1 as bait was employed. Upon interrogation of a mouse brain library, one gene product that interacts with VR1 and is highly homologous to human eferin was found. Its interaction with VR1 was confirmed by GST-pull-down and co-immunoprecipitation. When cotransfected into HEK cells, VR1 and eferin largely colocalize. Furthermore, in rat dorsal root ganglion cells, the rat eferin homologue also colocalizes with rVR1. However, this protein had no significant effect on VR1 channel activity in response to CAP. This was determined by two-electrode recording of oocytes and whole cell recording of HEK cells that were cotransfected with VR1 and human eferin. PMID- 15883037 TI - Acute and chronic effects of eicosapentaenoic acid on voltage-gated sodium channel expressed in cultured human bronchial smooth muscle cells. AB - This study investigated acute and chronic effects of eicosapentaenoic acid (EPA) on voltage-gated Na+ current (I(Na)) expressed in cultured human bronchial smooth muscle cells (hBSMCs). The whole-cell voltage clamp technique and quantitative real-time RT-PCR analysis were applied. The alterations in the fatty acid composition of phospholipids after treatment with EPA were also examined. Extracellular application of EPA produced a rapid and concentration-dependent suppression of tetrodotoxin-sensitive I(Na) with the half-maximal inhibitory concentration of 2 microM. After washing out EPA with albumin, I(Na) returned to the control level. Similar inhibitory effects were observed regarding other fatty acids (docosahexaenoic, arachidonic, stearic, and oleic acids), but EPA was the most potent inhibitor. The effect of EPA on I(Na) was not blocked by nordihydroguaiaretic acid and indometacin, and was accompanied by a significant shift of the steady-state inactivation curve to more negative potentials. In cells chronically treated with EPA, the EPA content of the cell lipid fraction (mol%) increased time-dependently, while arachidonic acid (AA) decreased, resulting in an increase of EPA to AA ratio. Then, the level of mRNA (SCN9A) encoding I(Na) decreased significantly. These results provide novel evidence that EPA not only rapidly inhibits I(Na), but also reduces the mRNA levels of the Na+ channel after cellular incorporation of EPA in cultured hBSMCs. PMID- 15883038 TI - Signaling through Disabled 1 requires phosphoinositide binding. AB - The Reelin signaling pathway plays a critical role in the correct positioning of neurons within the developing brain. Within this pathway, Disabled 1 (Dab1) serves as an intracellular adaptor that is tyrosine phosphorylated when Reelin, a secreted glycoprotein, binds to the lipoprotein receptors VLDLR and ApoER2 on the surface of neurons. The phosphotyrosine-binding (PTB) domain within its amino terminus enables Dab1 to recognize and bind to a conserved sequence motif within the cytoplasmic tails of the receptors. In addition, the PTB contains a Pleckstrin Homology-like subdomain that binds to phosphoinositides. Here, we show that the phosphoinositide-binding region within Dab1 PTB domain is required for membrane localization and basal tyrosine phosphorylation of Dab1 independently of VLDLR and ApoER2. Furthermore, receptor-independent membrane targeting of Dab1 is required for its interaction with Src and Crk, and disruption of phosphoinositide binding also blocks subsequent Reelin-induced tyrosine phosphorylation of Dab1. PMID- 15883039 TI - Therapeutic effects of PG201, an ethanol extract from herbs, through cartilage protection on collagenase-induced arthritis in rabbits. AB - In order to assess the therapeutic effects of PG201 (an ethanol extract from herbs) on osteoarthritis, we investigated whether PG201 could suppress the disease progression of collagenase-induced arthritis (CNIA) in rabbits. The right knees of rabbits were injected intra-articularly with collagenase, and the rabbits were orally treated with distilled water (DW), PG201 (200 mg/kg) or diclofenac (DCF, 10 mg/kg) once a day for 8 weeks. Oral administration of PG201 significantly suppressed the stiffness and bone space narrowing. Cartilage erosion and GAG release (p<0.01) were considerably reduced in the knee joints. As well, the mRNA expression of matrix degradation enzymes including MMP-1, -3, and 13 was decreased. On the contrary, the concentrations of TIMP-2 in the synovial fluids were considerably amplified in the PG201 treated group (p<0.01), but not in the DCF treated group. The pathologic inflammatory molecules involved in cartilage destruction such as IL-1beta, PGE2, and NO were also diminished by PG201. Taken together, these results indicate that PG201 has therapeutic effects on CNIA through the prominent protection of cartilage. PG201 indeed has great potential as a form of treatment for osteoarthritis. PMID- 15883040 TI - Bovine adenovirus type 3 internalization is independent of primary receptors of human adenovirus type 5 and porcine adenovirus type 3. AB - Usefulness of adenoviral vectors derived from human adenovirus (HAd) type 5 (HAd5) is mainly limited by wide prevalence of preexisting anti-HAd5 immunity as well as non-specific tissue tropism of these vectors. As an alternative, non human adenoviral vectors including bovine adenovirus type 3 (BAd3) are currently being investigated. Non-prevalence of BAd3 in humans and its ability to evade preexisting HAd immunity are some of the features that make BAd3 a promising vector for human gene delivery. BAd3 appears to have a tissue tropism distinct from that of HAd5 and also the repertoire of cells efficiently transduced by BAd3 is different. We performed antibody-mediated receptor blocking experiments to show that BAd3 internalization was independent of coxsackievirus-adenovirus receptor, the primary determinant of HAd5 tropism, or integrin alpha(v)beta3, a secondary molecule involved in HAd5 entry. Using homologous and heterologous knob mediated competition assays with recombinant knobs of HAd5, porcine adenovirus type 3 (PAd3), or BAd3, we observed that BAd3 internalization was independent of the primary receptors of HAd5 and PAd3. These results provide support for further exploration of BAd3 vectors for designing targeted vectors for human gene therapy. PMID- 15883041 TI - Retroviral display of urokinase-binding domain fused to amphotropic envelope protein. AB - Tumors frequently express urokinase (uPA) receptor (uPAR). To investigate whether uPAR can efficiently target cancerous cells using amphotropic retroviral vectors, we generated a retrovirus displaying the amino-terminal fragment (ATF) of uPA as an N-terminal extension of viral envelope protein. We also made use of a "two step strategy" by inserting a uPA cleavage site between the ATF moiety and the envelope. We measured the ability of ATF-bearing chimeric envelopes to infect huPAR-overexpressing Madin-Darby canine kidney (MDCK) and control MDCK II cells. The ATF-viruses infected both MDCK cell lines with an equivalent efficiency, suggesting that the chimeric viruses were not sequestered by uPAR and infect cells preferentially via the Pit-2 receptor. The addition of a uPA cleavage site increased the infection level of huPAR-MDCK cells by 2-fold when uPA was present in the infection medium. Surprisingly, ATF-env viruses infected huPAR-MDCK cells 5.5-fold more efficiently in the presence of exogenous uPA. This stimulatory effect of uPA on infection of huPAR-MDCK cells by the ATF-env virus was completely abolished by methyl-beta-cyclodextrin, suggesting that this effect involves the caveolar endocytosis pathway. PMID- 15883042 TI - Isolation and characterization of Dictyostelium thymidine kinase 1 as a calmodulin-binding protein. AB - Probing of a cDNA expression library from multicellular development of Dictyostelium discoideum using a recombinant radiolabelled calmodulin probe (35S VU1-CaM) led to the isolation of a cDNA encoding a putative CaM-binding protein (CaMBP). The cDNA contained an open reading frame of 951 bp encoding a 227aa polypeptide (25.5 kDa). Sequence comparisons led to highly significant matches with cytosolic thymidine kinases (TK1; EC 2.7.1.21) from a diverse number of species including humans (7e-56; 59% Identities; 75% Positives) indicating that the encoded protein is D. discoideum TK1 (DdTK1; ThyB). DdTK1 has not been previously characterized in this organism. In keeping with its sequence similarity with DdTK1, antibodies against humanTK1 recognize DdTK1, which is expressed during growth but decreases in amount after starvation. A CaM-binding domain (CaMBD; 20GKTTELIRRIKRFNFANKKC30) was identified and wild type DdTK1 plus two constructs (DdTK deltaC36, DdTK deltaC75) possessing the domain were shown to bind CaM in vitro but only in the presence of calcium while a construct (DdTK deltaN72) lacking the region failed to bind to CaM. Thus, DdTK1 is a Ca2+ dependent CaMBP. Sequence alignments against TK1 from vertebrates to viruses show that CaM-binding region is highly conserved. The identified CaMBD overlaps the ATP-binding (P-loop) domain suggesting CaM might affect the activity of this kinase. Recombinant DdTK is enzymatically active and showed stimulation by CaM (113+/-0.5%) an in vitro enhancement that was prevented by co-addition of the CaM antagonists W7 (91.2+/-0.8%) and W13 (96.6+/-0.6%). The discovery that TK1 from D. discoideum, and possibly other species including humans and a large number of human viruses, is a Ca2+-dependent CaMBP opens up new avenues for research on this medically relevant protein. PMID- 15883043 TI - The cyclin-dependent kinase 11 interacts with 14-3-3 proteins. AB - Cyclin-dependent kinase 11 isoforms (CDK11) are members of the p34(cdc2) superfamily. They have been shown to play a role in RNA processing and apoptosis. In the present study, we investigate whether CDK11 interacts with 14-3-3 proteins. Our study shows that the putative 14-3-3 binding site (113-RHRSHS-118) within the N-terminal domain of CDK11(p110) is functional. Endogenous CDK11(p110) binds directly to 14-3-3 proteins and phosphorylation of the serine 118 within the RHRSHS motif seems to be required for the binding. Besides, CDK11(p110) is capable of interacting with several different isoforms of 14-3-3 proteins both in vitro and in vivo. The interaction of 14-3-3 gamma with CDK11(p110) occurs throughout the entire cell cycle and reaches maximum at the G2/M phase. Interestingly, 14-3-3 gamma shows strong interaction with N-terminal portion of caspase-cleaved CDK11(p110) (CDK11(p60)) product at 48 h after Fas treatment, which correlates with the maximal cleavage level of CDK11(p110) and the maximum activation level of CDK11 kinase activity during apoptosis. Collectively, these results suggest that CDK11 kinases could be regulated by interaction with 14-3-3 proteins during cell cycle and apoptosis. PMID- 15883044 TI - Effect of dimethyl sulphoxide on the crystal structure of porcine pepsin. AB - The structure of porcine pepsin crystallized in the presence of dimethyl sulphoxide has been analysed by X-ray crystallography to obtain insights into the structural events that occur at the onset of chemical denaturation of proteins. The results show that one dimethyl sulphoxide molecule occupies a site on the surface of pepsin interacting with two of its residues. An increase in the average temperature factor of pepsin in the presence of dimethyl sulphoxide has been observed indicating protein destabilization induced by the denaturant. Significant increase in the temperature factor and weakening of the electron density have been observed for the catalytic water molecule located between the active aspartates. The conformation of pepsin remains unchanged in the crystal structure. However, the enzyme assay and circular dichroism studies indicate that dimethyl sulphoxide causes a slight change in the secondary structure and complete loss of activity of pepsin in solution. PMID- 15883045 TI - Alpha-tocopheryl succinate induces cytostasis and apoptosis in osteosarcoma cells: the role of E2F1. AB - Alpha-tocopheryl succinate (alpha-TOS), a redox-inactive analog of vitamin E, induces cell cycle arrest, differentiation, and triggers apoptosis. We examined the ability of alpha-TOS to induce cytostasis and/or apoptosis in two human osteosarcoma cell lines, which carry wild-type pRb but differ in the p53 status. In the wt-p53 cells, alpha-TOS induced apoptosis, which was associated with p53 activation and enhanced E2F1 expression. Mutant p53 cells failed to undergo apoptosis when challenged with alpha-TOS. The cell growth arrest after alpha-TOS treatment was associated with a reduced expression of E2F1. Knocking down E2F1 rendered the alpha-TOS-sensitive cells rather resistant to the apoptotic stimulus inducing a marked and prolonged cell growth arrest. We conclude that alpha-TOS induces cell growth arrest or apoptosis involving E2F1. PMID- 15883046 TI - The acetylcholine receptor gamma-to-epsilon switch occurs in individual endplates. AB - Maturation of the neuromuscular junction is accompanied by molecular switching of acetylcholine receptor (AChR) channels from embryonic types with gamma-subunits to adult ones with epsilon-subunits after birth. As a step toward understanding the molecular mechanisms of the gamma-to-epsilon switch, we addressed the question of whether embryonic- and adult-type AChRs constitute different endplates during the transitional period. From analyses with double- or triple staining with anti-gamma- and/or anti-epsilon-antibodies together with alpha bungarotoxin, which binds to alpha-subunits, we demonstrated that during neonatal stages in mice, adult-type AChRs are incorporated into individual endplates expressing embryonic-AChRs and replace these embryonic-AChRs gradually. The main period of AChR transition in the mouse diaphragm was between postnatal days 5 (P5) and P7, similar to the period described previously in which endplates shift from multi-axon to single-axon innervation. This finding will help our understanding of the mechanisms of the gamma-to-epsilon switch during establishment of the neuromuscular junction. PMID- 15883047 TI - Pregnane X receptor-agonists down-regulate hepatic ATP-binding cassette transporter A1 and scavenger receptor class B type I. AB - Pregnane X receptor (PXR) is the molecular target for a wide variety of endogenous and xenobiotic compounds. It regulates the expression of genes central to the detoxification (cytochrome P-450 enzymes) and excretion (xenobiotic transporters) of potentially harmful compounds. The aim of the present investigation was to determine the role of PXR in regulation of high-density lipoprotein (HDL) cholesterol metabolism by studying its impact on ATP-binding cassette transporter A1 (ABCA1) and scavenger receptor class B type I (SR-BI) expression in hepatocytes. ABCA1 and SR-BI are major factors in the exchange of cholesterol between cells and HDL. Expression analyses were performed using Western blotting and quantitative real time RT-PCR. Luciferase reporter gene assays were used to measure promoter activities. Total cholesterol was measured enzymatically after lipid extraction (Folch's method). The expression of ABCA1 and SR-BI was inhibited by the PXR activators rifampicin and lithocholic acid (LCA) in HepG2 cells and pregnenolone 16alpha-carbonitrile (PCN) in primary rat hepatocytes. Thus, PXR appears to be a regulator of hepatic cholesterol transport by inhibiting genes central to cholesterol uptake (SR-BI) and efflux (ABCA1). PMID- 15883048 TI - Functional analysis of sigH expression in Corynebacterium glutamicum. AB - The sigH gene of Corynebacterium glutamicum encodes ECF sigma factor sigmaH. The gene apparently plays an important role in other stress responses as well as heat stress response. In this study, we found that deleting the sigH gene made C. glutamicum cells sensitive to the thiol-specific oxidant diamide. In the sigH mutant strain, the activity of thioredoxin reductase markedly decreased, suggesting that the trxB gene encoding thioredoxin reductase is probably under the control of sigmaH. The expression of sigH was stimulated in the stationary growth phase and modulated by diamide. In addition, the SigH protein was required for the expression of its own gene. These data indicate that the sigH gene of C. glutamicum stimulates and regulates its own expression in the stationary growth phase in response to environmental stimuli, and participates in the expression of other genes which are important for survival following heat and oxidative stress response. PMID- 15883049 TI - Effects of small heat shock proteins on the thermal denaturation and aggregation of F-actin. AB - Effect of recombinant chicken small heat shock protein with molecular mass 24 kDa (Hsp24) and recombinant human small heat shock protein with molecular mass 27 kDa (Hsp27) on the heat-induced denaturation and aggregation of skeletal F-actin was analyzed by means of differential scanning calorimetry and light scattering. All small heat shock proteins did not affect thermal unfolding of F-actin measured by differential scanning calorimetry, but effectively prevented aggregation of thermally denatured actin. Small heat shock protein formed stable complexes with denatured (but not with intact) F-actin. The size of these highly soluble complexes was smaller than the size of intact F-actin filaments. It is supposed that protective effect of small heat shock proteins on the cytoskeleton is at least partly due to prevention of aggregation of denatured actin. PMID- 15883050 TI - Mutational and inhibitive analysis of SARS coronavirus 3C-like protease by fluorescence resonance energy transfer-based assays. AB - The 3C-like protease (3CL(pro)) of severe acute respiratory syndrome coronavirus (SARS-CoV) plays key roles in viral replication and is an attractive target for anti-SARS drug discovery. In this report, a fluorescence resonance energy transfer (FRET)-based method was developed to assess the proteolytic activity of SARS-CoV 3CL(pro). Two internally quenched fluorogenic peptides, 1NC and 2NC, corresponding to the N-terminal and the C-terminal autocleavage sites of SARS-CoV 3CL(pro), respectively, were used as substrates. SARS-CoV 3CL(pro) seemed to work more efficiently on 1NC than on 2NC in trans-cleavage assay. Mutational analysis demonstrated that the His41 residue, the N-terminal 7 amino acids, and the domain III of SARS-CoV 3CL(pro) were important for the enzymatic activity. Antibodies recognizing domain III could significantly inhibit the enzymatic activity of SARS CoV 3CL(pro). The effects of class-specific protease inhibitors on the trans cleavage activity revealed that this enzyme worked more like a serine protease rather than the papain protease. PMID- 15883051 TI - A novel role for myosin II in insulin-stimulated glucose uptake in 3T3-L1 adipocytes. AB - Insulin-stimulated glucose uptake requires the activation of several signaling pathways to mediate the translocation and fusion of GLUT4 vesicles from an intracellular pool to the plasma membrane. The studies presented here show that inhibition of myosin II activity impairs GLUT4-mediated glucose uptake but not GLUT4 translocation to the plasma membrane. We also show that adipocytes express both myosin IIA and IIB isoforms, and that myosin IIA is recruited to the plasma membrane upon insulin stimulation. Taken together, the data presented here represent the first demonstration that GLUT4-mediated glucose uptake is a myosin II-dependent process in adipocytes. Based on our findings, we hypothesize that myosin II is activated upon insulin stimulation and recruited to the cell cortex to facilitate GLUT4 fusion with the plasma membrane. The identification of myosin II as a key component of GLUT4-mediated glucose uptake represents an important advance in our understanding of the mechanisms regulating glucose homeostasis. PMID- 15883052 TI - Evaluation of EST-data using the genome assembly. AB - Using expressed sequence tag (EST) data for genomewide studies requires thorough understanding of the nature of the problems that are related to handling these sequences. We investigated how EST clustering performs when the genome is used as guidance as compared to pairwise sequence alignment methods. We show that clustering with the genome as a template outperforms sequence similarity methods used to create other EST clusters, such as the UniGene set, in respect to the extent ESTs originating from the same transcriptional unit are separated into disjunct clusters. Using our approach, approximately 80% of the RefSeq genes were represented by a single EST cluster and 20% comprised of two or more EST clusters. In contrast, approximately 25% of all RefSeq genes were found to be represented by a single cluster for the UniGene clustering method. The approach minimizes the risk for overestimations due to the amount of disjunct clusters originating from the same transcript. We have also investigated the quality of EST-data by aligning ESTs to the genome. The results show how many ESTs are not adequately trimmed in respect of vector sequences and low quality regions. Moreover, we identified important problems related to ESTs aligned to the genome using BLAT, such as inferring splice junctions, and explained this aspect by simulations with synthetic data. EST-clusters created with the method are available upon request from the authors. PMID- 15883053 TI - Single inner cell masses yield embryonic stem cell lines differing in lifr expression and their developmental potential. AB - The unique differentiation potential of inner cell mass derived embryonic stem cells together with their outstanding self-renewal capacity makes them a desirable source for somatic cell therapy of human diseases. Somatic cells are gained by in vitro differentiation of embryonic stem cells, however, the differentiation potential of embryonic stem cells varied even between isogenic cell lines. Variable differentiation potentials may either be a consequence of an inherent inhomogeneity of gene expression in the inner cell mass or may have technical reasons. To understand variations in the differentiation potential, we generated pairs of isogenic, monozygotic twin, and single inner cell mass derived clonal embryonic stem cell lines, and demonstrate that they differentially express the leukaemia inhibitory factor receptor gene. Variations of leukaemia inhibitory factor receptor protein levels are already evident in the inner cell mass and predispose the cardiomyogenic potential of embryonic stem cell lines in a Janus activated kinase dependent manner. Thus, a single inner cell mass may give rise to embryonic stem cell lines with different developmental potentials. PMID- 15883054 TI - Insertion/deletion polymorphism in clusterin gene influences serum lipid levels and carotid intima-media thickness in hypertensive Japanese females. AB - Clusterin has been implicated in lipid metabolism and atherogenesis, however, the influence of genetic variation has not been examined in Japanese. In this study, we identified 11 single nucleotide polymorphisms (SNPs) of clusterin gene by direct sequencing. Among them, one promoter SNP (-4453T>G), one missense SNP (4183G>A), and 2 common SNPs (5608T>C and 6316delT) were genotyped in 525 asymptomatic hypertensives not treated with lipid lowering agents. -4453T>G, 4183G>A, and 5608T>C showed no correlation with the clinical characteristics, however, in the 6316delT, an insertion (I)/deletion (D) polymorphism, D/D subjects had significantly higher levels of total cholesterol and low-density lipoprotein (LDL)-cholesterol than I/I subjects in females but not in males. Female subjects with the D allele (D/D+I/D) had greater intima-media thickness of the carotid artery than I/I subjects. In a multiple logistic regression analysis, the D allele of 6316delT was detected as an independent predictor for the plaque prevalence. In conclusion, the clusterin gene polymorphism may contribute to the serum lipid levels and the progression of carotid atherosclerosis in hypertensive Japanese females. PMID- 15883057 TI - Transthoracic discectomy without interbody fusion. AB - BACKGROUND: Transthoracic discectomy is an established surgical procedure for the treatment of thoracic disk disease. Most authors advocate interbody fusion after transthoracic discectomy. The purpose of this study was to determine if there were any adverse consequences in foregoing interbody fusion after transthoracic discectomy. METHODS: Eighteen consecutive patients underwent transthoracic discectomy without fusion between 1996 and 2002 at Mayo Clinic (Rochester, MN). There were 11 women and 7 men with the mean age of 54 years (range, 28-84 years). Surgical indications were radiculopathy in 1 patient and myelopathy in 17. Follow up data were obtained from the clinic visits and telephone surveys. We used the available pre- and postoperative radiographs for 16 patients at the last follow up to establish the incidence of postoperative kyphosis and/or scoliosis at the operated level. Mean duration of the radiographic follow-up was 22 +/- 24 (SD) months. RESULTS: None of the patients reported the onset of a new axial spine pain postoperatively. No patient developed segmental kyphosis or scoliosis at the operated level during the follow-up period. Fifteen of 18 (83%) patients had significant improvement in their neurological symptoms and signs. Two patients remained unchanged. An 83-year-old patient had a slight worsening of her gait after surgery. Specifically, the only 3 nonambulatory patients regained ambulation after discectomy. There were 4 complications: 1 wound infection, 1 pleural effusion requiring pleurodesis, 1 cerebrospinal fluid leak, and 1 case of disabling intercostal neuralgia. CONCLUSIONS: These results indicate that interbody fusion may not be necessary for selected patients undergoing transthoracic discectomy. Further long-term follow-up is needed to evaluate the development of late spinal instability and resultant deformity after this procedure. PMID- 15883059 TI - Missile-caused median nerve injuries: results of 81 repairs. AB - BACKGROUND: Missile-caused median nerve injuries have rarely been reported in current literature. We present repair outcome for all median nerve injuries in which the median nerve was severed either by missile injury or secondarily in the subsequent resection of a neuroma in continuity. METHODS: Prospective study included 81 casualties with proximal, intermediate, or distal complete median nerve lesions, repaired by nerve graft or direct suture. Final outcome was defined at least 4 years postoperatively, on the basis of motor recovery, sensory recovery, neurophysiological recovery, and patient's judgment on the outcome, each estimated by 0 to 5 points and according to the total sum, as poor, insufficient, good, or excellent. The last 2 modalities were considered to be successful. RESULTS: Successful outcome was obtained in 68.7% of distal, in 33.3% of intermediate, and in 10% of proximal repairs. Average outcomes were good, insufficient, and poor, respectively (P < .001). Nerve defect and preoperative interval were both significantly shorter on average for patients with successful outcome than for those with unsuccessful outcome (P < .001 and P = .007, respectively), but only preoperative interval and height of repair were independent predictors for successful outcome. Age of patient, associated ulnar nerve complete lesion, and manner of repair did not influence the outcome significantly (P > .05). CONCLUSIONS: The level of repair, duration of preoperative interval, and length of nerve defect significantly influence outcome after median nerve repair, but only level of repair and duration of preoperative interval were independent predictors for successful outcome. PMID- 15883061 TI - Influence of head position on the effectiveness of twist drill craniostomy for chronic subdural hematoma. AB - BACKGROUND: Twist drill craniostomy with closed system drainage (TDC-CSD) is a well-accepted treatment of chronic subdural hematomas (CSDH). Although this intervention has a long track record of effectiveness, little is known of its relationship with the head position of the patient (flat vs elevated). This study evaluated if the position of the patient's head influences outcome. METHODS: The database of a University Hospital Center was queried for patients who had CSDH treated by TDC-CSD between January 1997 and March 2001. Identified patients were grouped into 2 categories: head of bed (HOB) at 30% and HOB flat while undergoing treatment. Outcomes were then evaluated with regard to amount of drainage, complications, recurrence, and length of hospital stay (LOS). RESULTS: Forty-four patients were identified who received TDC-CSD treatment of CSDH. Of these, 24 patients had flat HOB and 20 had HOB elevated to 30 degrees . Although patients with elevated HOB had higher amounts of drainage (239 vs 166 mL), this figure did not reach statistical significance (P = .23). The number of recurrences and complications likewise did not reach statistical significance. Despite these findings, a statistically significant difference in LOS was found between the groups (flat = 5.5 days, elevated = 8.1 days, P = .03). This was believed secondary to bias resulting from placing the HOB of healthier patients (based on Glasgow Coma Scale) flat. CONCLUSIONS: Elevation of the patient's head during TDC CSD treatment of CSDH does not seem to impact the amount of drainage, recurrence frequency, or complication rate. Although a statistically significant difference in LOS was observed based on this variable, it appeared to be the result of bias in patient selection for HOB elevation. PMID- 15883063 TI - Intracranial aneurysms in the pediatric population: case series and literature review. AB - BACKGROUND: Intracranial aneurysms in children (< or =18 years old) are rare, and their epidemiology is poorly understood. We present a prospective 14-year experience at our institution and review the literature to clarify the characteristics and outcomes of pediatric patients with intracranial aneurysms. METHODS: Review of medical records in the Johns Hopkins aneurysm prospective database and review of the literature since 1939 were performed. RESULTS: Of 1377 intracranial aneurysm cases at our institution from 1991 to 2004, 19 (1.4%) pediatric patients were treated using microsurgical or endovascular techniques. Male/female ratio was 2.2:1. Eleven percent occurred at the internal carotid artery (ICA) bifurcation, and 42% were located in the posterior circulation. Seven (37%) were giant lesions; 58% presented with subarachnoid hemorrhage. Patients in good Hunt and Hess grade (I-III) comprised 42%, and poor-grade (IV-V) patients comprised 16%. At a mean follow-up duration of 3.0 +/- 3 years, 95% of patients demonstrated favorable outcomes with Glasgow Outcome Scale score of 4 or 5. In the literature, a total of 706 pediatric intracranial aneurysm cases have been described since 1939. The male/female ratio is 1.8:1. The ICA bifurcation was the location in 26%, and only 17% were in the posterior circulation. Twenty percent were giant lesions, and 80% presented with subarachnoid hemorrhage. Good- and poor-grade patients comprise 49% and 36% of the total cases, respectively. Favorable outcome was reported in 60%. CONCLUSIONS: During recent decades, good outcomes after the treatment of ruptured and unruptured pediatric aneurysms have increased, reaching 95% in the current series. Pediatric intracranial aneurysms occur more commonly in male patients and have a predilection for the terminal ICA bifurcation. PMID- 15883065 TI - Electromyographic investigation on handling forces of mechanically counterbalanced and sensor-servomotor-supported surgical microscopes. AB - BACKGROUND: Recent developments in sensor-servomotor-driven microscopes again initiated a discussion on the value of such technology for practical use in neurosurgery. The need for less force in moving a motor-supported microscope is advantageous. However, questions remain if well-known difficulties in the past such as resonance phenomenon, loss of natural feeling, and unequal handling forces in different situations have been overcome by the new generation of sensor servo-supported surgical microscopes. METHODS: Handling forces of a mechanically counterbalanced neurosurgical microscope (Zeiss NC4, Zeiss, Oberkochen, Germany) were compared with those of a sensor-servo-supported neurosurgical microscope (Moeller HiR 20-1000, Moeller, Wedel, Germany). Handling forces were correlated with the surface electromyogram measurement of the muscle activity of 4 neurosurgeons. The activity of the forearm muscles was measured while handling the 2 different microscopes in standardized tests. RESULTS: The electrophysiologic measurement revealed that significantly less muscle activity was required to handle the sensor-servo-driven microscope in all directions. The untrained surgeons profited less than the skilled ones. Differences were most evident with disbalanced microscopes. CONCLUSIONS: With this technology, the neurosurgeons exerted less effort, especially in strenuous test situations where the single-handed use of the microscope was mandatory. The reduced muscle forces that move the sensor-servo-type microscope and the continuously balanced state that might help prevent unwanted correction movements will ease intraoperative handling in general. PMID- 15883068 TI - Brain abscess: clinical experience and analysis of prognostic factors. AB - BACKGROUND: Over the past 2 decades, the diagnosis and treatment of brain abscess have been facilitated by a number of technological advancements, which have resulted in a significant improvement of outcome. The aim of this manuscript is to review our experience, to determine the factors related to the outcome, and to improve the therapeutic strategy for this disease. METHODS: From 1986 to 2002, 178 consecutive patients with bacterial brain abscess were treated at the National Taiwan University Hospital, Taipei, Taiwan. We reviewed their clinical presentation, bacteriology, treatment, and outcome retrospectively. Groups were compared by chi2 test, Fisher exact test, or t test as appropriate. Multivariate logistic regression with backward selection was used to select the set of covariates that were independently associated with outcome. RESULTS: One hundred eleven patients (62%) had favorable outcome, 14 patients (8%) had severe disability, 9 patients (5%) became vegetative, and 44 (25%) died during hospitalization. Patients with better Glasgow Coma Scale (GCS) on admission, no underlying disease, positive culture, or surgical treatment were more likely to have a good outcome. Patients with nasopharyngeal carcinoma, acquired immunodeficiency syndrome, hematologic disease, deep-seated abscess, or medical treatment alone were more likely to have a poor outcome. Multivariate analysis revealed that only GCS, immunodeficiency, and presence of underlying disease related with outcome. CONCLUSIONS: The poor prognostic factors of brain abscess are poor GCS, immunodeficiency, and presence of underlying disease. Aggressive treatment with surgery when indicated and careful management of specimen for culture might improve outcome. PMID- 15883071 TI - Microanatomy of the subependymal arteries of the lateral ventricle. AB - BACKGROUND: Scarce information about the anatomy of the subependymal arteries (SEAs) is present in the scientific literature. METHODS: Twenty cerebral hemispheres with injected arteries were microdissected, and the magnetic resonance imaging scans of 100 patients with lacunar infarcts were examined. RESULTS: The SEAs were found to range in diameter from 40 to 490 microm (mean, 149 microm) and in number between 3 and 12 (average, 5.2). Of these, numbers from 1 to 3 originated from the anterior choroidal artery (AChA), between 1 and 10 from the lateral posterior choroidal artery (LPChA), 1 from the medial posterior choroidal artery (MPChA), and 1 from the internal carotid artery. The SEAs most often arose from the choroidal branches (90%) and less frequently from the thalamic (30%), caudate (35%), or thalamocaudate twigs (20%). The SEAs of the AChA supplied the walls of the temporal horn (100%), the occipital horn (85%), and the atrium (35%). Those of the LPChA perfused the walls of the occipital horn (15%), the atrium (65%), the body of the ventricle (100%), and partially the frontal horn. The SEAs of the MPChA partially nourished the body and the frontal horn (10%). The SEAs may also occasionally supply the caudate nucleus (20%) and the stria terminalis. The anastomoses involving the SEAs were absent. In spite of this, ischemia in the territory of a single SEA was noticed in only 1% of our patients. CONCLUSIONS: The SEAs are tiny vessels that supply the walls of the lateral ventricle, as well as the caudate nucleus and the stria terminalis occasionally. The obtained anatomic data can have important neurosurgical implications in intraventricular operations. PMID- 15883073 TI - Management of Rathke's cleft cysts. AB - BACKGROUND: Incidental diagnosis of Rathke's cleft cysts (RCCs) has increased due to the improvements in imaging techniques; however, symptomatic cases are rare and accurate preoperative diagnosis can be difficult. METHODS: Files and magnetic resonance imaging (MRI) of 10 surgically treated patients with RCC and 8 conservatively managed cases with a suspected diagnosis were reviewed retrospectively. Clinical, radiological, and histological features as well as intraoperative findings were scrutinized. RESULTS: In the surgical group, headache was present in 4 patients, hormonal abnormality in 4, visual deficits in 3, and otolaryngological symptoms in 2. Two of the nonsurgical cases had hyperprolactinemia, and the remaining were asymptomatic. The cyst had suprasellar extension (SSE) in 9 patients and was entirely suprasellar in 1 in the surgical group. It was purely intrasellar in 6 nonsurgical cases and had SSE in 2. The cyst content showed hyperintense MRI signal in the majority of T1 and T2 images in both groups. Transsphenoidal surgery was performed in 8 and transcranial surgery in 2 patients. Squamous metaplasia and inflammation was present in 3 cases each. Initial hormonal deficiencies did not improve postoperatively and new deficits were observed in 2 cases. There was no recurrence with an average follow up of 32 months. CONCLUSIONS: Rathke's cleft cysts is a rare pathology with a wide spectrum of clinical and radiological features. Reactive inflammation of the normal pituitary gland may have a role in the pathogenesis of hormonal deficiency, in addition to compression. Preoperative recognition of the anteriorly displaced normal residual gland may be important in avoiding postoperative hormonal deficiency after transsphenoidal approach. PMID- 15883076 TI - Effect of cell phone magnetic fields on adjustable cerebrospinal fluid shunt valves. AB - The rapid increase in the number of cell phone users has led to the suggestion that electromagnetic waves might affect medical devices. Cerebrospinal fluid shunt valves contain a magnetic device to allow the intracranial pressure setting to be adjusted transcutaneously. Among the valves tested, the settings of the Strata valve, the Hakim valve, and the Sophy valve were affected by magnetic flux densities of 6.0, 17.5, and, 40.0 mT, respectively. Cell phones produce a magnetic flux density of 3.0 to 40.0 mT. Although cell phones could theoretically influence shunt valves, this seems unlikely because the flux density decreases with the square of the distance. PMID- 15883077 TI - Combination treatment of vertebral metastases using image-guided percutaneous radiofrequency ablation and vertebroplasty: a case report. AB - BACKGROUND: We describe a case of vertebral metastases treated with a combination of percutaneous radiofrequency ablation (RFA) and vertebroplasty in a single session. CASE DESCRIPTION: A 45-year-old female with recently diagnosed metastatic non-small cell lung cancer was referred for consideration of vertebroplasty after having several weeks of back pain. Computed tomography and magnetic resonance imaging scans of the thoracic spine revealed metastatic lesions at the T9 and T12 bodies. Radiofrequency ablation was performed using a starburst array electrode (Rita Medical Systems, Mountain View, Calif). The metastatic lesion in the T9 vertebral body measured 1.5 x 1.5 cm and was heated to 100 degrees C for 5 minutes. Vertebroplasty was then performed on this lesion as well as a lesion at T12 with no complications. The patient was discharged home the same day without complaints. CONCLUSION: Radiofrequency ablation and vertebroplasty are minimally invasive procedures that can be used in combination to treat spinal metastases. The pain relief from these treatments is often immediate. RFA can also be used to minimize procedure-related complications during the vertebroplasty. In properly selected patients, RFA may also increase the duration of local spinal stabilization afforded by the vertebroplasty alone. Long-term studies are warranted to assess the efficacy and durability of this unique approach. PMID- 15883080 TI - Cerebellar mutism in adults after posterior fossa surgery: a report of 2 cases. AB - BACKGROUND: Mutism has been associated with injury to midline cerebellar structures secondary to degenerative disease, tumors, hemorrhage, or surgery. Typically, cerebellar mutism syndrome (CMS) has been seen in children and only rarely described in adults after surgery of the posterior fossa. This syndrome typically arises 48 hours after the initiating event and resolves approximately 7 to 8 weeks later. Characteristics of CMS include complete absence of speech without impaired consciousness, other cranial nerve deficits, or long tract signs. CASE DESCRIPTION: The authors report on 2 patients each of whom developed cerebellar mutism after tumor resection using a posterior fossa approach. The first patient underwent gross total resection of a pineal region tumor via a supracerebellar approach. The second patient underwent posterior fossa decompression for a left cerebellar hemispheric renal cell carcinoma metastasis with adjacent hemorrhage. One patient displayed a variant of cerebellar mutism with severe ataxic dysarthria without complete absence of speech, whereas the other demonstrated frank mutism. After neuroimaging studies confirmed the absence of a surgically treatable postoperative cause for the patients' symptoms, they were managed in a supportive fashion (eg, speech therapy) and improved within 3.5 months and 1 year, respectively. CONCLUSION: It is paramount that neurosurgeons be aware of cerebellar mutism with regard to its very rare occurrence in adults, its time of onset, and typical self-limiting course. PMID- 15883081 TI - An adult case of moyamoya syndrome that developed dural sinus thrombosis associated with protein C deficiency: case report and literature review. AB - We describe a 54-year-old woman exhibiting MMS who developed delayed dural sinus thrombosis associated with PCD. Angiographic findings of the patient were so unusual that bilateral internal carotid arteries were occluded between their origin and the carotid fork with extensive development of collateral circulation via the external carotid arteries and the posterior cerebral arteries instead of moyamoya vessels at the base of the brain. Seven years after bilateral cerebral revascularization surgery, intracerebral hemorrhage occurred caused by dural sinus thrombosis. In the treatment for the patient with MMS associated with PCD, risk of sinus thrombosis should be taken into account. PMID- 15883084 TI - Experience from "double-insurance bypass." Surgical results and additional techniques to achieve complex aneurysm surgery in a safer manner. AB - BACKGROUND: "Double-insurance bypass" was recently advocated to avoid the risk of cerebral ischemia during prolonged temporary occlusion of the carotid artery. For large aneurysms needing temporary but prolonged obliteration of the internal carotid artery (ICA). We have attempted the double-insurance bypass in 15 patients and, herein, report the efficacies and limitations of the procedure, and surgical techniques to make this procedure safer. METHODS: We treated 15 patients with complex internal carotid aneurysms by clipping surgery with the aid of radial artery (RA) to proximal middle cerebral artery (MCA) bypass, so-called double-insurance bypass. We analyzed surgical results of the procedure. RESULTS: In 11 patients, the duration of temporary occlusion of the ICA could be prolonged for as long as 110 minutes (mean, 45 minutes) without any ischemic complications. One patient in the earlier period of our experience suffered extended cerebral infarction due to possible restricted blood flow through the RA, because the brachial artery was compressed by the firm shoulder joint and neighboring structures. Thereafter, we routinely monitored the blood pressure of MCA (MCABP) and never experienced such cortical infarctions. Another 3 patients, however, experienced ischemia in the territory of perforating arteries that originated from a segment that could not be perfused by the RA-MCA bypass. CONCLUSIONS: In combination with monitoring of MCABP, the double-insurance bypass can be a safer and more potent adjunctive procedure for the treatment of complex internal carotid aneurysms which require prolonged temporary occlusion of the ICA. PMID- 15883086 TI - "Is this trip necessary?". PMID- 15883087 TI - Acute and subchronic effects of lead on the central and peripheral nervous systems in rats in combination with alcohol. AB - Humans are exposed, either simultaneously or sequentially, to various chemicals, including the neurotoxicants lead and ethanol. The aim of the present work was to investigate the changes in the spontaneous cortical activity (electrocorticogram; ECoG) and in the stimulus-dependent evoked potentials (EPs) recorded from rats pre-treated with alcohol and treated with lead acutely (intraperitoneally) or subchronically (by gavage). The measured parameters were spectral composition of the ECoG, amplitude and the latency of the stimulus-evoked cortical potential, as well as compound action potential amplitude, conduction velocity, and relative and absolute refractory period in a peripheral nerve. With subchronic lead and alcohol treatment, significant increase in the frequency of spontaneous activity and slight decrease in the EP amplitude were seen. In acute administration, EP amplitude increased and conduction velocity of the tail nerve decreased significantly. Our results showed that, in a combined exposure situation which is likely to happen also in humans, the known effects of neurotoxic heavy metals can be more severe. PMID- 15883088 TI - Influence of metal exposure history on trace metal uptake and accumulation by marine invertebrates. AB - This paper reviews recent studies on the influence of trace metal exposure history on subsequent metal uptake in marine invertebrates, particularly bivalves and barnacles. Trace metal exposure may induce specific metal-binding ligands. Metallothionein-like proteins appear to play an important role in mediating metal uptake and hence accumulation, especially for Cd. Other ligands such as sulfide are important for Ag biokinetic changes in bivalves. The incorporation of trace metals into metal-rich granules appears to have little if any effect in modifying trace metal uptake. Furthermore, metals also interact strongly in their accumulation by marine animals. Generally, dissolved Hg uptake is reduced following exposure to other metals such as Ag, Cd, Cu, and Zn. Preexposure of marine mussels to Zn also causes concomitant changes in the uptake of other metals such as Cd sharing similar transport pathways. In general, barnacles are less responsive than mussels to trace metal exposure, presumably because of the dominance of detoxificatory pathways in which the accumulated metals are bound in granules (relatively biologically inactive pools). The tissue body burden and the detoxificatory fate of metals in animals seem to be more important in affecting metal accumulation than the nature of the exposure routes (aqueous vs. dietary) or of the exposure regimes. Trace metal accumulation may also be variable in different natural populations of bivalves as a result of different physicochemical environments and histories of exposure. We finally suggest a few areas for future research in this field. PMID- 15883089 TI - Body residues: a key variable to analyze toxicity tests with Chironomus riparius exposed to copper-spiked sediments. AB - Fourth instar Chironomus riparius larvae were exposed to four sediments spiked with copper. Length and copper concentration in the tissue were monitored daily. Kinetics data analysis was performed by fitting a one-compartment model. The analysis of growth data was performed using an energy-based approach which estimates a no-effect concentration (NEC). The elimination rate of copper did not depend on the sediment characteristics, whereas the bioconcentration factor differed by a factor of 10 among the tested sediments, accounting for differences in copper bioavailability. Consequently, the NEC expressed in term of exposure concentrations differed substantially between sediments. In contrast, the NEC expressed in terms of body residues did not depend on the sediment characteristics. This work links, for the first time, a mechanistic effects model with the critical body residue approach, and could contribute to develop relevant tools for sediment risk assessment. PMID- 15883090 TI - Selenium impacts on razorback sucker, Colorado: Colorado River III. Larvae. AB - Razorback sucker (Xyrauchen texanus) larvae from adults exposed to selenium at three sites near Grand Junction, Colorado, for 9 months were used in a 30-day waterborne and dietary selenium study. Selenium concentrations in water averaged <1.6 microg/L from 24-Road, 0.9 microg/L from Horsethief, 5.5 microg/L from Adobe Creek, and 10.7 microg/L from the North Pond. Selenium in dietary items averaged 2.7 microg/g in brine shrimp, 5.6 microg/g in zooplankton from Horsethief east wetland, 20 microg/g in zooplankton from Adobe Creek, and 39 microg/g in zooplankton from North Pond. The lowest survival occurred in larvae fed zooplankton rather than brine shrimp. Survival of larvae at Adobe Creek and North Pond was lower in site water than in reference water. Survival of brood stock larvae was higher than Horsethief larvae even though they received the same water and dietary treatments. Arsenic concentrations in brine shrimp may have resulted in an antagonistic interaction with selenium and reduced adverse effects in larvae. Deformities in larvae from North Pond were similar to those reported for selenium-induced teratogenic deformities in other fish species. Selenium concentrations of 4.6 microg/g in food resulted in rapid mortality of larvae from Horsethief, Adobe Creek, and North Pond, and suggested that selenium toxicity in the Colorado River could limit recovery of this endangered fish. PMID- 15883091 TI - Reduced growth and survival of larval razorback sucker fed selenium-laden zooplankton. AB - Four groups of larval razorback sucker, an endangered fish, were exposed to selenium-laden zooplankton and survival, growth, and whole-body residues were measured. Studies were conducted with 5, 10, 24, and 28-day-old larvae fed zooplankton collected from six sites adjacent to the Green River, Utah. Water where zooplankton were collected had selenium concentrations ranging from <0.4 to 78 microg/L, and concentrations in zooplankton ranged from 2.3 to 91 microg/g dry weight. Static renewal tests were conducted for 20 to 25 days using reference water with selenium concentrations of <1.1 microg/L. In all studies, 80-100% mortality occurred in 15-20 days. In the 28-day-old larvae, fish weight was significantly reduced 25% in larvae fed zooplankton containing 12 microg/g selenium. Whole-body concentrations of selenium ranged from 3.7 to 14.3 microg/g in fish fed zooplankton from the reference site (Sheppard Bottom pond 1) up to 94 microg/g in fish fed zooplankton from North Roadside Pond. Limited information prior to the studies suggested that the Sheppard pond 1 site was relatively clean and suitable as a reference treatment; however, the nearly complete mortality of larvae and elevated concentrations of selenium in larvae and selenium and other elements in zooplankton indicated that this site was contaminated with selenium and other elements. Selenium concentrations in whole-body larvae and in zooplankton from all sites were close to or greater than toxic thresholds where adverse effects occur in fish. Delayed mortality occurred in larvae fed the two highest selenium concentrations in zooplankton and was thought due to an interaction with other elements. PMID- 15883092 TI - Do seasonal changes affect metallothionein induction by metals in mussels, Mytilus edulis? AB - Mussels have been proposed as biomonitors of metal pollution based on the determination of metallothionein (MT) concentrations as a biomarker, but a comprehensive study taking into account both intersite and long-term temporal variations in MT and metal concentrations in different organs is lacking. Thus, the present study was designed to examine the concentrations of cytosolic and insoluble Cd, Cu, Zn, and MT in gills and digestive gland of mussels (Mytilus edulis) of homogeneous size and age obtained from aquaculture and kept on a reference site or translocated to a metal-rich site throughout their reproductive season (March-October 1997). Relatively significant binding of metals to the insoluble fraction was observed in both tissues. In the digestive gland, monthly MT concentrations were strongly correlated to cytosolic metal levels. Moreover, despite significant temporal variations, the grand mean MT concentration based on all individual determinations in the digestive gland (reference, n=54; transplants, n=50) was significantly higher in mussels from the metal-rich site. On the other hand, gill MT concentrations did not reflect metal contamination as reliably. PMID- 15883093 TI - Oxidative stress and genotoxic responses to resin acids in Mediterranean mussels. AB - This study represents the first attempt to investigate the genotoxic effects and oxidative stress of resin acids in Mediterranean mussels (Mytilus galloprovincialis Lmk). Mussels were exposed to 2.7 microM abietic acid (AA) and dehydroabietic acid (DHAA) for 6, 12, 18, and 24h. Gill and hepatopancreas conjugation activity, antioxidant defense system, lipid peroxidation (LPO), and DNA damage were determined as reduced glutathione (GSH), glutathione S transferase (GST) activity, glutathione peroxidase (GPx) activity, catalase (CAT) activity, LPO, and DNA strand breaks. AA caused significant GST inhibition in mussel gills at 12, 18, and 24h. Activity of the antioxidant enzymes, namely, GPx and CAT, was inhibited at 24 and 18 h, respectively, in mussel gills. A significant increase in gill LPO was observed at 24h. The DNA integrity of mussel hepatopancreas significantly decreased after 12 and 24 h exposure to AA. A significant increase in LPO was observed after 6h exposure to DHAA, in either mussel gills or hepatopancreas. DNA integrity was significantly decreased in mussel hepatopancreas after 12 and 24 h exposure to DHAA. AA induced oxidative damage and genotoxicity in mussels, because it promoted increases in LPO in gills and DNA strand breaks in hepatopancreas. DHAA promoted oxidative damage and genotoxicity in mussels, as significant increases were observed in LPO in gills and hepatopancreas and in DNA strand breaks in hepatopancreas. PMID- 15883094 TI - Anguilla anguilla L. liver EROD induction and genotoxic responses after retene exposure. AB - Anguilla anguilla L. (European eel) were exposed for 8, 16, 24, and 72 h to 0 (control), 0.1, 0.3, 0.9, and 2.7 microM retene. A. anguilla L. liver ethoxyresorufin-O-deethylase EROD activity significantly increased during the whole exposure period to all retene concentrations, when compared to their controls. The liver cytochrome P450 content only increased after exposure to high retene concentrations (0.9 and 2.7 microM) from 8 to 24 and 72 h, respectively. Generally, liver DNA integrity decreased with increased retene concentrations. Thus, a low retene concentration (0.1 microM) was only effective at 16 h, 0.3 and 0.9 microM had an early and prolonged effect up to 24h, and 2.7 microM decreased liver DNA integrity during the whole exposure period. However, blood DNA integrity decrease was observed in eels after 24h exposure to 0.1 microM retene, and at 16 h to 0.3 and 0.9 microM retene, despite an early blood DNA integrity decrease at 8 and 16 h exposure to 2.7 microM retene. An early genotoxic response to retene was also observed as erythrocyte nuclear abnormalities plus Notched (ENA+Not) frequency increase at 8, 16, and 24h exposure to 0.1 and 0.3 microM retene as well as at 8, 16, 24, and 72 h to 0.9 microM retene. Though, the highest retene concentration (2.7 microM) only induced ENA+Not and erythrocyte nuclear abnormalities minus Notched (ENA-Not) at 16 h exposure. The eel ENA+Not increase was more sensitive than the ENA-Not increase as a measure of retene genotoxicity. Eel liver alanine amino transferase (ALT) increased activity reveals its enhanced transamination capacity after short-term exposure to retene. The A. anguilla L. ratio between hemoglobin concentration and red blood cells count (Hb/RBC) increased at 8h exposure to 0.1, 0.3, and 0.9 microM retene, suggesting an initial homeostatic process. PMID- 15883095 TI - Assessing the genotoxicity of imidacloprid and RH-5849 in human peripheral blood lymphocytes in vitro with comet assay and cytogenetic tests. AB - A combined approach employing comet assay and micronucleus (MN) and sister chromatid exchanges (SCE) tests was utilized to assess the genotoxicity of two pesticides, imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitro-imidazolidin-2 ylideneamine] and RH-5849 [2'-benzoyl-1'-tert-butylbenzoylhydrazine], on human peripheral blood lymphocytes in vitro. No significant difference in the frequencies of MN and SCE from the negative groups (P>0.05) was observed at low dose levels (i.e., 0.05 mg/L for imidacloprid and 5mg/L for RH-5849). As the concentrations of imidacloprid and RH-5849 were increased to 0.1 and 25 mg/L, respectively, significant effects to the frequencies of MN and SCE (P<0.05) were achieved relative to those of the negative controls. MN and SCE frequencies increased similarly in a dose-related manner with both pesticides. With the comet assay, however, the distribution of DNA damage grades in all the pesticide treated groups was significantly different from those in the control (P<0.01). DNA damage scores increased with the exposure levels of both pesticides, and linear dose-effect relationships were observed for both imidacloprid (r2=0.98) and RH-5849 (r2=0.92). The cytogenetic techniques and comet assay revealed potential adverse effects of both imidacloprid and RH-5849 in human peripheral blood lymphocytes in vitro. Combination of the comet assay and cytogenetic tests appears commendable to assess the potential risks of human exposure to the pesticides. PMID- 15883096 TI - Health variables and gill morphology in the tropical fish Astyanax fasciatus from a sewage-contaminated river. AB - The relative condition factor (Kn), gonadosomatic index (GSI), selected hematological variables and gill morphology of the fish Astyanax fasciatus were analyzed in two sites (site 1 was unpolluted and site 2 was polluted with untreated domestic sewage) of a tropical river (Camanducaia river, Sao Paulo State, Brazil). The relationship between the body mass (M(B)) and the standard length (L(S)) of A. fasciatus from both sites was M(B)=0.00799 L(S)(3.51843). The Kn values from both males and females and the GSI of females were significantly higher in site 2. The mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were higher in females from site 2. Gill tissue anomalies and gill parasites were rare in fish from both sites; however, the number of chloride cells was significantly higher in fish from site 1. A. fasciatus presents high capacity to live in ion-poor and soft water and is able to compensate for environmental changes caused by untreated domestic sewage discharges. PMID- 15883097 TI - Bioaccumulation of mercury in muscle tissue of fish in the Elbe River (Czech Republic): multispecies monitoring study 1991-1996. AB - The study monitored mercury (Hg) contamination of fish muscle tissue at 13 geographical sites that can be regarded as crucial points for an ecotoxicological assessment of the Czech Republic section of the Elbe River. The descriptive part of the study was primarily aimed at comparative evaluation of the Hg load during the period 1991-1996. The conclusions were supported by multivariate statistical analyses of the content of Hg in the muscle tissue of 1251 fish belonging to 23 species with four dominant indicator species: Perca fluviatilis (n=163), Abramis brama (n=173), Rutilus rutilus (n=148), and Leuciscus cephalus (n=166). Considering data from 3- to 5-year-old fish, significantly increased contamination was detected in typical predators compared to the other fish species in all sites (P<0.001). On the other hand, omnivorous and planctivorous species were ranked as the least sensitive for Hg pollution. Perch appeared to be the most contaminated species in the sample with muscle Hg concentration in the range of 0.840-1.398 mg Hg kg(-1). Although less contaminated than perch, muscle contamination of bream sensitively separated differently contaminated sites; the highest load ranged from 0.368 to 0.543 mg Hg kg(-1). Time-related comparison of sampling campaigns revealed no significant trend changes, in either sediment samples or fish tissue. Thus, the analyses documented an evidently rather stabilized total Hg pollution in the Elbe River environment. Multivariate multispecies analyses found the age of analyzed individuals and the feeding strategy of a given species as the most important, however mutually interactive, covariates for Hg accumulation in muscle tissue. The analyses revealed decreasing sensitivity of older predator individuals to differentiate highly and moderately contaminated sites. Benthophagous species mostly kept their discrimination capacity toward contaminated sites in all age categories, with the exception of bream that was rather linked to the pattern typical for predator species. The unclear position of omnivorous species, represented namely by roach, corresponded with their weak bioindicator power, mainly in the young age categories. PMID- 15883098 TI - Toxicity of nickel, zinc, and cadmium to nitrate uptake in free and immobilized cells of Scenedesmus quadricauda. AB - We examined the influences of three trace metals on the accumulation of a major nutrient (NO3-) in Scenedesmus quadricauda. A comparative study on metal-nutrient interaction in free and immobilized states of algal cells was conducted. The effect due to interaction between different variables (cell state type, metal type, and metal dose) was studied to assess the variation in the nitrate uptake by free and immobilized cells. The results analyzed by ANOVA (three-way) (components: cell state type, metal type, and metal dose) confirmed that the inhibition of nitrate uptake by test metals was highly significant (P<0.001). Free and immobilized states of S. quadricauda responded differently (P<0.05, ANOVA) to the types of metal added. Uptake kinetics was studied by monitoring short-term uptake rates at different nutrient levels. Free and immobilized cells of the organism displayed noncompetitive modes of inhibition for Ni and Zn while a competitive mode of inhibition by Cd was observed in both free and immobilized states of the organism. PMID- 15883099 TI - Aquatic toxicity of nitrogen mustard to Ceriodaphina dubia, Daphnia magna, and Pimephales promelas. AB - Investigation of toxicity of mustard compounds to aquatic organisms has been limited although their effects on terrestrial mammal species have been well studied. In this study, the 48-h LC50 values of nitrogen mustard (HN2) are reported for two aquatic invertebrate species (Daphnia magna and Ceriodaphnia dubia) and for one fish species (Pimephales promelas). Mean LC50 values to C. dubia, D. magna, and P. promela were 1.12, 2.52, and 98.86 mg/L, respectively. C. dubia was the species most sensitive to HN2. Seven-day lethal and sublethal tests with P. promelas and C. dubia were also conducted. In chronic tests, fathead minnow growth was significantly reduced by 2.50 mg/L HN2, while C. dubia reproduction was significantly affected by 7.81 mug/L HN2. These adverse effects on aquatic organisms caused by lower-level concentrations of HN2 indicate that a possible aquatic ecosystem disaster could occur either after a chemical spill or during chemical warfare. PMID- 15883100 TI - Use of sodium dodecyl sulfate and zinc sulfate as reference substances for toxicity tests with the mussel Perna perna (Linnaeus, 1758) (Mollusca: Bivalvia). AB - Effects of anthropogenic pollution have been observed at different trophic levels in the oceans, and toxicity tests constitute one way of monitoring these alterations. The present assay proposes the use of two reference substances, sodium dodecyl sulfate (SDS) and zinc sulfate, for Perna perna larvae. This common mussel on the Brazilian coast is used as a bioindicator and is of economic interest. The chronic static embryo-larval test of short duration (48 h) was employed to determine the NOEC, LOEC, and IC50 for SDS and zinc sulfate, as well as the coefficient of variation. Salinity, pH and un-ionized ammonia (NH3) and dissolved oxygen (DO) concentrations were measured to monitor water quality. The results demonstrated that the main alterations in veliger larvae are the development of only one shell, protruded mantle, malformed shell, formation of only part of a valve, clipped edges, uneven sizes and presence of a concave or convex hinge. NOEC values were lower than 0.25 mg L(-1) for zinc sulfate and 0.68 mg L(-1) for SDS. The coefficient of variation was 17.63% and 2.50% for zinc sulfate and SDS, respectively. PMID- 15883101 TI - Women's health after WHI. PMID- 15883102 TI - Practical guidelines for postmenopausal hormone therapy. AB - The fourth Amsterdam Menopause Symposium (2-4 October 2004) was dedicated to practical recommendations to guide clinicians after the confusion, concerns, and controversies generated by study results over the previous several years. Those recommendations are summarized in this deliberately concise and user-friendly document, always recognizing that each clinician must help women with their decision-making according to individual needs, desires, and understanding of benefits and risks. PMID- 15883103 TI - Climacteric medicine: European Menopause and Andropause Society (EMAS) 2004/2005 position statements on peri- and postmenopausal hormone replacement therapy. AB - In women experiencing distressing climacteric symptoms during the peri- and postmenopause there is conclusive evidence from abundant randomised controlled trials that systemic hormone replacement therapy (HRT) of any type affords symptom relief, with no alternative treatment producing similar effect. Though this evidence is accumulating, the question of how to provide best clinical practice in an attempt to both alleviate the menopausal symptoms and prevent the more long-term postmenopausal degenerative diseases is still under debate. When providing climacteric medicine, the dose and regimen of HRT needs to be individualised based on the principle of choosing the lowest appropriate dose in relation to severity of symptoms and on the menopausal age. However, few long term data on different HRT formulations exist in symptomatic women, which also account for baseline risk of cardiovascular disease (CVD), breast cancer and osteoporosis. In most cases, an individualized prescription together with life style management will sustain possibilities for net beneficial effects on climacteric symptoms, quality of life (QoL), sexuality and osteoporosis, with only rare risk of severe adverse effects. With the perspective provided by recent epidemiological findings, not least from the estrogen only arm of the Women's Health Initiative Study (WHI), European Menopause and Andropause Society (EMAS) supports research activities in symptomatic women with new HRT formulations in order to affect positively the balance of clinical benefit and risk, including specific information on QoL and also account for the traditional differences in treatment modalities between the US and Europe, and the difference in BMI, life style and diet. In women experiencing an early menopause (<45 year) current data support a specific overall benefit of HRT. At present, more long-term systemic HRT may be considered in women at high risk of osteoporotic fractures, in particular when alternate therapies are either inappropriate or insufficiently effective, as benefits will outweigh any risks. In contrast, urogenital symptoms may be addressed efficiently and safely with long-term local estrogen therapy. PMID- 15883104 TI - Guidelines for hormone treatment of women in the menopausal transition and beyond. PMID- 15883105 TI - Tibolone: clinical recommendations and practical guidelines. A report of the International Tibolone Consensus Group. AB - An international multidisciplinary panel of experts in the management of the menopause met at the 4th Amsterdam Menopause Symposium in October 2004 to determine the specific place of tibolone, a synthetic steroid with a unique clinical profile, within the wide range of currently available postmenopausal therapy options. The consensus was that tibolone is a valuable treatment option for women with climacteric complaints. As well as relieving vasomotor symptoms, tibolone has positive effects on sexual well-being and mood, and improves vaginal atrophy and urogenital symptoms. Prevention of bone loss with tibolone is comparable to that seen with estrogen therapy (ET) and estrogen/progestogen therapy (EPT). As tibolone rarely causes endometrial proliferation, no additional progestogen is required. It also has good tolerability, being associated with a low incidence of vaginal bleeding and of breast pain. Tibolone does not increase mammographic density. Absolute numbers of women at increased risk for breast cancer are estimated to be low or absent with both tibolone and ET, and the risk with tibolone should be significantly lower than that with EPT. Tibolone might therefore be preferable to EPT in certain women who have not been hysterectomised. Based on the evidence available, the panel proposed a number of subgroups of postmenopausal women with vasomotor symptoms in whom tibolone might have added value; these included women with sexual dysfunction, mood disorders, fibroids and urogenital complaints, as well as those with breast tenderness or high mammographic breast density with EPT use. PMID- 15883106 TI - The woman patient after WHI. AB - An epidemic of fear and distrust has infected women (and physicians) after publication of the Women's Health Initiative (WHI). The overinflated negative data emerging from the oestroprogestinic arm of WHI have frightened women and gave rise to the most difficult emotions to cope with. Keywords such as cancer, death and hormones combined together, have potentiated an avoidant attitude towards hormonal therapy (HT) driven more emotionally than rationally. This negative aura has not been dissipated by the positive data from the oestrogen only arm of WHI. This paper will discuss: women's different emotional reactions to and coping strategies for HT-related fear after WHI; the communication skills physicians should use in focusing on positive messages emerging from WHI; predictors of current HT use; the meaning of the higher use of HT in postmenopausal highly-educated women and women gynaecologists; the importance of increasing healthy life-styles as a taking of responsibility towards aging by every woman; shifting from passivity to active sharing of the decision making process with the caring physician; and the use of an individually tailored HT, when appropriate, as part of an active strategy in the pursuit of a longer health expectancy. PMID- 15883107 TI - Menopause 2005: transatlantic confrontation of opinions. PMID- 15883108 TI - Menopause and HRT--the state of the art in Europe. AB - The HRT field has been dramatically affected by the publication on major randomised controlled trials of the long-term effects of HRT. The publicity surrounding the publication of these data has affected public and regulatory perceptions of HRT and its role in healthcare, including the relatively short term use of HRT for the relief of menopausal symptoms. An evidence-based appraisal of the role of HRT today is best achieved by considering the different components of the effects of HRT individually, based on the best trial evidence, and then considering these together in the context of the age of woman concerned and the specific components relevant to that woman's health profile. This paper summarises the effects of HRT using this approach in the context of European practice today and describes the events surrounding the regulatory and scientific society position statements. PMID- 15883109 TI - WHI and aftermath: looking beyond the figures. AB - In contrast to the preliminary results of the WHI conjugated equine estrogen plus medroxyprogesterone acetate arm published 2 years ago, the final results from that arm as well as preliminary data from the conjugated equine estrogen-only arm showed that the panic over hormone treatment for menopausal women was unjustified. Moreover, the data for women younger than 60-years-old was even more reassuring. WHI was a study on elderly women starting hormones under the assumption that it may confer cardioprotection. The results of WHI should not be generalized and should not be a reason to withhold hormones in symptomatic perimenopausal or early post-menopausal women who could benefit from that therapy. PMID- 15883110 TI - Coronary heart disease in menopausal women: implications of primary and secondary prevention trials of hormones. AB - In direct contrast to the observational studies, both primary and secondary prevention trials of female reproductive hormones have found no benefit for coronary heart disease (CHD). Basic science studies have elucidated several mechanisms by which estrogen may improve coronary arterial physiology and prevent pathology, but have also found mechanisms by which estrogen might increase coagulation or inflammation, or might trigger coronary events in advanced lesions. Animal studies suggest that hormones may retard early atherosclerosis, while both animal studies and human angiographic trials are conclusive that hormones do not retard progression of raised lesions. Hormone use in the primary prevention observational studies would mostly have started at the age of menopause, in women whose arteries on average would be closer to normal than those of women in the clinical trials. One hypothesis worthy of further study is that estrogen may have a beneficial effect in normal or near-normal arteries, but the opposite effect in the presence of established atherosclerosis. However, at the average age of menopause, a substantial proportion of women has raised lesions, and a smaller proportion already has advanced lesions. Also, the apparent benefit of hormone use was found in secondary prevention observational studies, i.e., in women with compromised arteries. It is likely that uncorrected biases in the observational studies lead to an overestimation of any benefit of hormone use. On the other hand, endogenous estradiol may be responsible for the later onset of coronary disease in women compared to men; if so, then the appropriate test of the estrogen hypothesis would employ transdermal estradiol in a young population of menopausal women. Hormones are not indicated for the prevention of CHD, particularly in the light of the increased risk for stroke and venous thrombosis. Their use for other indications (menopausal symptoms, osteoporosis) needs to be tempered by the risk for cardiovascular disease (CVD). PMID- 15883111 TI - Controversies about HRT--lessons from monkey models. AB - Lessons from monkey models contribute significantly to a better understanding of the controversies in reconciling the differences in postmenopausal hormone treatment outcomes between observational and randomized trial data. Monkey studies brought attention to premenopausal estrogen deficiency with resulting premature coronary artery atherosclerosis. Recently, those monkey studies were confirmed for premenopausal women in the NHLBI-sponsored Women's Ischemia Syndrome Evaluation (WISE) Study. Monkey studies have provided convincing evidence for the primary prevention of coronary artery atherosclerosis when estrogens are administered soon after the development of estrogen deficiency. Equally convincing are the data from monkey studies indicating the total loss of these estrogens beneficial effects if treatment is delayed for a period equal to six postmenopausal years for women. An attempt has been made using the monkey model to identify the hormone treatment regimen most effective in preventing the progression of coronary artery atherosclerosis. By a substantial margin, the most effective approach is that of using estrogen containing oral contraceptive during the perimenopausal transition, followed directly by hormone replacement therapy postmenopausally. Because of similarities between human and nonhuman breast, monkeys have had a major role in clarifying controversies surrounding the breast cancer risk of estrogen only versus estrogen plus progestin therapies. The results of monkey studies suggest little or no effects of estrogen only treatment; whereas, estrogen+progestin clearly increases breast cancer risk. PMID- 15883112 TI - Postmenopausal hormone therapy and breast cancer: what is the problem? AB - Observational studies provide evidence that breast cancer risk is increased with long-term oral use of postmenopausal estrogen replacement therapy (ET). Various large cohort studies have shown that the addition of a progestogen in combined hormone replacement therapy (EPT) increases this risk further. Prospective, randomized controlled trials have confirmed this for the continuous combined regimen. So, why not tell our patients, "Stop using ET and EPT, it is dangerous to your health!"? The answer is: there are too many problems to allow such an oversimplified, definite statement. What is the problem? There is more than one! The problems are as follows: In conclusion, we have a problem as we cannot formulate any general advice that holds for the majority of European postmenopausal women due to lack of consistency, lack of biological plausibility, and lack of relevance of randomized clinical trial data to our daily practical work. So, we have a problem and not a firm basis for undisputable statements. PMID- 15883113 TI - Breast cancer risk in the WHI study: the problem of obesity. AB - In the climacteric, about 40% of the women have occult breast tumors the growth of which may be stimulated by hormones. Many genetic, reproductive and lifestyle factors may influence the incidence of breast cancer. Epidemiological data suggest that the increase in the relative risk (RR) of breast cancer induced by hormone replacement therapy (HRT) is comparable with that associated with early menarche, late menopause, late first birth, alcohol consumption, etc. One of the most important risk factors is obesity which exceeds the effect of HRT by far, and in overweight postmenopausal women the elevated risk of breast cancer is not further increased by HRT. As in the WHI study the majority of women was overweight or obese, this trial was unsuitable for the investigation of breast cancer risk. In the women treated with an estrogen/progestin combination, the RR of breast cancer rose only in those women who have been treated with hormones prior to the study, suggesting a selection bias. In the women not pretreated with hormones, it was not elevated. In the estrogen-only arm of the WHI study, there was no increase but a steady decrease in the RR of breast cancer during 6.8 years of estrogen therapy. This result was unexpected, as estrogens are known to facilitate the development and growth of breast tumors, and the effect is enhanced by the addition of progestins. Obese women are at high risk to develop a metabolic syndrome including insulin resistance and hyperinsulinemia. In postmenopausal women, elevated insulin levels are not only associated with an increased risk for cardiovascular disease, but also for breast cancer. This might explain the effects observed in both arms of the WHI study: HRT with relative low doses of estrogens may improve insulin resistance and, hence, reduce the elevated breast cancer risk in obese patients, whereas this beneficial estrogen effect may be antagonized by progestins. The principal options for the reduction of breast cancer risk in postmenopausal women are the prevention of overweight and obesity to avoid the development of hyperinsulinemia, the medical treatment of insulin resistance, the use of low doses of estrogens and the reduction of exposure to progestins. The latter might include long-cycles with the sequential use of appropriate progestins every 3 months for 14 days. There are large inter individual variations in the proliferative response to estrogens of the endometrium. Control by vaginalsonography and progestin challenge tests may help to identify those women who may be candidates for low-dose estrogen-only therapy. PMID- 15883114 TI - Appropriate use of hormones should alleviate concerns of cardiovascular and breast cancer risk. AB - Since the publication of several recent randomized trials in the United States, prescriptions for hormonal therapy have dropped precipitously. This has been due, in large part, to the concerns about the increased risk of cardiovascular (CV) disease and breast cancer among the hormone users. This review takes the perspective that the appropriate use of hormones largely alleviates these concerns. The appropriate use of hormones pertains to treating younger, healthy women who have menopausal symptoms as well as using low-doses of hormones. In the randomized trials, suggesting an increased CV risk, the older women were largely asymptomatic and had other CV risk factors. Data are presented to suggest that there is no increased CV risk with hormonal therapy in younger, healthy women within 5 years of menopause. Moreover, a model is presented to attempt to explain the potential of preventing CV disease when estrogen is begun early, and the relative hazard associated with later use. The risk of breast cancer with hormonal therapy is put into perspective with the realization that this risk is related to hormonal dose and duration of use, and that the absolute risk remains small. Use of progestogens, in particular, appears to enhance this risk. The appropriate use of hormones also pertains to using lower-doses. Here data are presented showing efficacy with lower-doses and improved safety. With the use of lower-doses of estrogens, the progestogen dose, as required in women with a uterus, can be minimized. PMID- 15883115 TI - Time course analysis of closed- and open-loop grasping of the Muller-Lyer illusion. AB - The authors investigated whether the early or later stages of closed-loop (CL) and open-loop (OL) grasping movements were differentially influenced by the Muller-Lyer (ML) illusion. Participants (N = 21) reached out and grasped small (5 cm) and large (7 cm) objects embedded within fins-in and fins-out ML configurations. Grasping time (GT) was normalized, and absolute grip aperture (GA) as well as scaled illusion effects were computed at 20%, 40%, 60%, and 80% of GT. The results indicated that CL trials were refractory to the illusory array (i.e., from 20% to 80% of GT), whereas OL trials were influenced by the ML figure during that same time. Those findings suggest that CL trials were supported by unitary and metrical visual information, whereas OL trials were entirely supported by perception-based visual information. PMID- 15883116 TI - Movement-related sensory feedback mediates the learning of a new bimanual relative phase pattern. AB - On the basis of findings emphasizing the role of perceptual consequences in movement coordination, the authors tested the hypothesis that the learning of a new bimanual relative phase pattern would involve the matching of the movement related sensory consequences (rather than the motor outflow commands) to the to be-learned pattern. Two groups of participants (n = 10 in each) practiced rhythmically moving their forearms with a phase difference of 30 degrees . In 1 group, a difference in the arms' eigenfrequencies was imposed such that synchronous generation of the left and right motor commands resulted in the required relative phase (30 degrees ), yielding incongruence between the motor commands and their sensory consequences. In the other group, the experimenter imposed no eigenfrequency difference so that the sensory consequences were congruent with the motor commands. Throughout the practice period, performance of both groups was assessed repeatedly for the congruent situation (i.e., no eigenfrequency difference). On those criterion tests, both groups performed the required pattern equally well. The authors discuss that result, which corroborated the hypothesis, from a dynamical systems perspective. PMID- 15883117 TI - Quantification of manual force control and tremor. AB - Isometric impulse frequencies associated with active tremor and force regulation were examined in 10 patients with idiopathic Parkinson's disease (PD) and in 10 older adults (OAs) who performed an isometric tracing task. The authors decoupled and analyzed the data to determine whether PD-related tremor in the thumb and in the index finger during isometric force control are related and whether PD impairs the performance of volitional force control beyond the errors contributed by tremor. After decoupling, there were clear and robust differences in PD patients' control of isometric force that could not be attributed to action tremor error. Those errors, which occurred in the absence of movement, suggest impairment in coordinated recruitment and derecruitment of motor units during a fine-motor task. PMID- 15883118 TI - Observational learning: effects of bandwidth knowledge of results. AB - The authors investigated whether bandwidth knowledge of results (KR) during observation of a model's performance enhances motor skill learning. Following a pretest, 2 groups of participants (N = 28) observed a model practicing a timing task. The bandwidth group received KR about the model's performance only when his performance fell outside the criteria for a correct response. The yoked group received KR on the same trials as the bandwidth group did but were not told that the KR was only about incorrect performances. In that way, the authors avoided a confound between bandwidth and relative frequency effects on performance and learning. Following the observation phase, both groups of participants performed 10-min and 24-hr retention tests. Bandwidth KR enabled that group to reduce its performance variability and, to a lesser extent, to enhance its performance accuracy. The authors discuss the results with respect to the powerful effect of qualitative KR through observation. PMID- 15883119 TI - Effects of velocity and limb loading on the coordination between limb movements during walking. AB - The authors investigated the effects of velocity (increasing from 0.5 to 5.0 km/hr in steps of 0.5 km/hr) and limb loading on the coordination between arm and leg movements during treadmill walking in 7 participants. Both the consistency of the individual limb movements and the stability of their coordination increased with increasing velocity; the frequency coordination between arm and leg movements was 2:1 at the lower velocities and 1:1 at the higher velocities. The mass manipulation affected the individual limb movements but not their coordination, indicating that a stable walking pattern was preserved. The results differed qualitatively from those obtained in studies on bimanual interlimb coordination, implying that the dynamical principles identified therein are not readily applicable to locomotion. PMID- 15883120 TI - Support for an explanation of the guidance effect in motor skill learning. AB - The authors investigated whether the knowledge of results (KR) schedule influences the extent to which intrinsic feedback is noticed and used. Fifty-six participants received KR that was either delayed over 2 trials (Delay-2) or provided directly after each trial (Delay-0) during 160 trials of an unfamiliar aiming task. No-KR retention tests were given after 80 trials and 1 min and 24 hr after the end of acquisition. After retention, all participants were questioned about their use of intrinsic feedback during practice and whether those sources changed as a function of practice. The Delay-2 group performed significantly less accurately on the 1st and last blocks of acquisition trials but showed a significantly smaller performance decline from acquisition to retention. Moreover, the Delay-2 group noticed and used a greater variety of intrinsic feedback sources and its members were more likely to report that their usage changed with practice. PMID- 15883121 TI - Dual-task training reduces impact of cognitive task on postural sway. AB - Postural sway increases when a cognitive task is performed concurrently with a postural task. The author examined the hypothesis that following dual-task training, a concurrent cognitive task would not amplify postural sway. Participants (N = 18) were assigned to no-training, single-task training, or dual task training groups. Single-task training consisted of 3 sessions in which the postural task, quiet standing on a compliant surface, and the cognitive task, counting backward by 3s, were practiced separately. Dual-task training consisted of 3 sessions of concurrent practice of the cognitive and postural tasks. After training, performance of a concurrent cognitive task increased postural sway in the no-training and single-task training groups but not in the dual-task training group. Results suggest that dual-task practice improves dual-task performance. PMID- 15883122 TI - Learning the pedalo locomotion task. AB - The authors examined the learning function of a multiple biomechanical degrees of freedom coordination task. Four adult participants practiced the pedalo locomotion task for 350 trials over 7 days. On the basis of the Cauchy theorem, the authors applied a movement pattern difference score that provides a measure of convergence to a fixed point as the criterion for quantifying learning. The findings showed a significant reduction of the movement pattern difference score over practice. Neither an exponential (0.11) nor a power law (0.10) function accommodated a large percentage of the variance of the pattern difference measure on individual learning functions, but the respective fits were higher, although not different, for movement time (.57, .55). Principal components analysis showed a decrease of components over practice; the analysis also showed that 3-5 components were required to accommodate 90% of the variance of the whole-body motion at the end of the final practice session. Those findings on the learning functions for movement and outcome scores are discussed in relation to the redundancy of the biomechanical system in moving to a dynamical stable fixed point in this task. PMID- 15883123 TI - Oncostatin M in combination with tumour necrosis factor {alpha} induces a chondrocyte membrane associated aggrecanase that is distinct from ADAMTS aggrecanase-1 or -2. AB - OBJECTIVE: To determine whether oncostatin M (OSM) + tumour necrosis factor alpha (TNFalpha) induces aggrecanase activity in chondrocyte membranes, to determine the effects of transforming growth factor beta1 (TGFbeta1), interleukin 4 (IL4), and tissue inhibitor of metalloproteinases (TIMPs) on this activity, and to determine whether this activity is due to a known ADAMTS aggrecanase. METHODS: Aggrecanase activity and ability of agents to prevent membrane associated aggrecanase activity were assessed by Western blotting. Expression of known aggrecanases was measured by real time polymerase chain reaction in bovine nasal and human articular chondrocytes. RESULTS: Chondrocyte membrane associated aggrecanase activity and increased mRNA expression of ADAMTS-1, -4, -5, and -9, but not ADAMTS-4 or -15, were enhanced after stimulation by OSM+TNFalpha in bovine chondrocytes. This activity was inhibited by TIMP-3. In human chondrocytes, OSM+TNFalpha also enhanced ADAMTS-1 and -4 expression, but not that of other ADAMTSs. TNFalpha alone induced ADAMTS-9 expression, whereas OSM addition caused suppression. Both TGFbeta1 and IL4 blocked membrane associated aggrecanase activity and decreased OSM+TNFalpha-induced expression of ADAMTS-9 in bovine and human chondrocytes. IL4 down regulated ADAMTS-4 mRNA, whereas TGFbeta1 increased this expression in both bovine and human chondrocytes. CONCLUSIONS: OSM+TNFalpha up regulates membrane associated aggrecanase activity and several ADAMTS aggrecanase mRNAs in chondrocytes. The chondroprotective effects of IL4 and TIMP-3 suggest that they may have therapeutic benefit for aggrecanolysis, whereas the differential inhibitory effects of TGFbeta1 may limit its therapeutic potential. Induced membrane associated aggrecanase activity is distinct from known soluble ADAMTS aggrecanases and merits further investigation. PMID- 15883125 TI - False-positive findings on [18F]FDG-PET caused by non-neoplastic cellular elements after neoadjuvant chemoradiotherapy for non-small cell lung cancer. AB - We report two patients with non-small cell lung cancer who had a pathologically complete response after neoadjuvant chemoradiotherapy, although they had positive [(18)F]fluoro-deoxyglucose positron emission tomography ([(18)F]FDG-PET) scans. They underwent concurrent chemoradiotherapy, which resulted in a partial response determined by computed tomography (CT). While [(18)F]FDG-PET after chemoradiotherapy was positive, pathological examination showed that the tumors were fibrotic lesions with infiltration of lymphocytes and macrophages, with the appearance of metaplastic epithelial cells. The reasons for the false-positive results on [(18)F]FDG-PET were considered to be the high uptake of FDG in non neoplastic inflammatory cellular elements, i.e. macrophages, lymphocytes and metaplastic epithelial cells, and squamous metaplasia induced by chemoradiotherapy. Although several studies demonstrated that [(18)F]FDG-PET could predict the response of neoadjuvant treatment of non-small cell lung cancer, one should bear in mind that false-positive results could be observed in pathological complete response of non-small cell lung cancer after neoadjuvant chemoradiotherapy. PMID- 15883124 TI - Correlation of systemic superoxide dismutase deficiency to airflow obstruction in asthma. AB - RATIONALE: Increased oxidative stress and decreased superoxide dismutase (SOD) activity in the asthmatic airway are correlated to airflow limitation and hyperreactivity. We hypothesized that asthmatic individuals with higher levels of oxidative stress may have greater loss of SOD activity, which would be reflected systemically in loss of circulating SOD activity and clinically by development of severe asthma and/or worsening airflow limitation. METHODS: To investigate this, serum SOD activity and proteins, the glutathione peroxidase/glutathione antioxidant system, and oxidatively modified amino acids were measured in subjects with asthma and healthy control subjects. RESULTS: SOD activity, but not Mn-SOD or Cu,Zn-SOD protein, was lower in asthmatic serum as compared with control, and activity loss was significantly related to airflow limitation. Further, serum SOD activity demonstrated an inverse correlation with circulating levels of 3-bromotyrosine, a posttranslational modification of proteins produced by the eosinophil peroxidase system of eosinophils. Exposure of purified Cu,Zn SOD to physiologically relevant levels of eosinophil peroxidase-generated reactive brominating species, reactive nitrogen species, or tyrosyl radicals in vitro confirmed that eosinophil-derived oxidative pathways promote enzyme inactivation. CONCLUSION: These findings are consistent with greater oxidant stress in asthma leading to greater inactivation of SOD, which likely amplifies inflammation and progressive airflow obstruction. PMID- 15883126 TI - Improving the evidence base for promoting quality and equity of surgical care using population-based linkage of administrative health records. AB - This paper highlights the uses of population-based linkage of administrative health records to improve the quality, safety, and equity of surgical care. The primary focus of the paper is on the transfer of this type of research into policy and practice. In the modern era of evidence-based medicine, it is essential that not only is new evidence incorporated into clinical practice, but that the implementation and associated costs are monitored; this requires the setting of appropriate benchmarking criteria. Furthermore, it is imperative that all members of the population receive optimal health care and people are not discriminated against because of socio-economic, locational, or racial factors. The use of data linkage can assist with examining these aspects of health care and this paper provides real-life examples such as costs and adverse events from laparoscopic cholecystectomy, event monitoring for post-operative venous thrombosis, and inequalities in cancer care. The influence of these studies on clinical practice and policy is also discussed. Furthermore, this paper discusses the strengths and weaknesses of data linkage research and how to avoid pitfalls. Health researchers, clinicians, and policy-makers will find the discussion of these issues useful in their everyday practice. PMID- 15883127 TI - Injection use in two districts of Pakistan: implications for disease prevention. AB - OBJECTIVE: To estimate the annual number of injections per person in Sindh province of Pakistan and to describe their distribution with regard to prescribers, settings, and safety. DESIGN: A population-based cross-sectional study in July-September 2001. SETTING: Lyari, an urban town in Karachi district; and Digri, a rural subdistrict in Mirpur Khas district. STUDY PARTICIPANTS: We selected a population-based cluster sample of 1150 individuals aged > or =3 months. We interviewed one person per household for the number of encounters they had with health care providers, number and types of injections received, safety circumstances, and cost of injections during the past 3 months. Main outcome measure. The number of injections per person per year. RESULTS: After adjusting for age and sex, 68% of participants had received at least one injection in the previous 3 months (13.6 injections/person/year). The majority of the respondents received injections at the clinics of qualified general practitioners (n = 571, 67%) by dispensers (644, 76%). Most of the injections (n = 3446, 96%) were for curative purposes. A freshly opened syringe was used for only 454 (53%) of the injections. The average fee for receiving an injection was Rs. 51 (US$ 0.8). CONCLUSION: Injections are overused in Pakistan's Sindh province and the ratios of injection per capita that we found are among the highest ever reported. INTERVENTIONS: are needed to substantially reduce injection prescription among private health care providers who prescribe most of the injections received by the population. PMID- 15883128 TI - Quality of primary health care in Saudi Arabia: a comprehensive review. AB - OBJECTIVES: Little is known about the quality of primary care in Saudi Arabia, despite the central role of primary care centers in Saudi health strategy. This study presents an overview of quality of primary care in Saudi Arabia, and identifies factors impeding the achievement of quality, with the aim of determining how the quality of Saudi primary care could be improved. METHOD: Using a systematic search strategy, data were extracted from the published literature on quality of care in Saudi primary care services, and on barriers to achieving high-quality care. RESULTS: Of the 128 studies initially identified, 31 met the inclusion criteria for the review. Studies identified were diverse in methodology and focus. Components of quality were reviewed in terms of access and effectiveness of both clinical and interpersonal care. Good access and effective care were reported for certain services including: immunization, maternal health care, and control of epidemic diseases. Poor access and effectiveness were reported for chronic disease management programs, prescribing patterns, health education, referral patterns, and some aspects of interpersonal care including those caused by language barriers. Several factors were identified as determining whether high-quality care was delivered. These included management and organizational factors, implementation of evidence-based practice, professional development, use of referrals to secondary care, and organizational culture. CONCLUSION: There is substantial variation in the quality of Saudi primary care services. In order to improve quality, there is a need to improve the management and organization of primary care services. Professional development strategies are also needed to improve the knowledge and skills of staff. PMID- 15883129 TI - The craving withdrawal model for alcoholism: towards the DSM-V. Improving the discriminant validity of alcohol use disorder diagnosis. AB - AIMS: To compare the discriminant validity of the DSM-IV and the ICD-10 classification of alcohol use disorders (AUD) with an alternative classification, the craving withdrawal model (CWM). CWM requires craving and withdrawal for the diagnosis of alcohol dependence and raises the alcohol abuse threshold to two DSM IV AUD criteria. METHODS: Data were derived from The Netherlands Mental Health Survey and Incidence Study, a large representative sample of the general Dutch population. In the present study, only non-abstinent subjects were included (n=6041). Three diagnostic systems (DSM-IV, ICD-10, and CWM) were compared using the following discriminant variables: alcohol intake, psychiatric comorbidity, functional status, familial alcohol problems, and treatment sought. RESULTS: The year prevalence of CWM alcohol dependence was lower than the prevalence of ICD-10 and DSM-IV dependence (0.3% vs 1.4% and 1.4%). The year prevalence of abuse was similar for CWM and DSM-IV (4.7 and 4.9%), but lower for ICD-10 harmful use (1.7%). DSM-IV resulted in a poor distinction between normality and abuse and ICD 10 resulted in a poor distinction between harmful use and dependence. In contrast, the CWM distinctions between normality and abuse, and between abuse, and dependence were significant for most of the discriminant variables. CONCLUSION: This study indicates that CWM improves the discriminant validity of AUD diagnoses. The predictive validity of the CWM for alcohol and other substance use disorders remain to be studied. PMID- 15883130 TI - Physical constraints on temperature difference in some thermogenic aroid inflorescences. AB - BACKGROUNDS AND AIMS: Thermogenesis in reproductive organs is known from several plant families, including the Araceae. A study was made of the relationship between temperature increase and spadix size in the subfamily Aroideae in order to determine whether the quantitative variation of heat production among species and inflorescences of different sizes follows a physical law of heat transfer. * METHODS: Spadix temperature was measured in 18 species from eight genera of tropical Araceae from the basal clade of Aroideae, both in French Guiana and in the glasshouses of the Montreal Botanical Garden. * KEY RESULTS: A significant logarithmic relationship was found between the volume of the thermogenic spadix zone and the maximum temperature difference between the spadix and ambient air. Four heat transfer models were applied to the data (conductive heat transfer alone, convective heat transfer alone, radiative heat transfer alone, and convective and radiative heat transfers) to test if physical (geometric and thermic) constraints apply. Which heat transfer model was the most probable was determined by using the criterion of a classical minimization process represented by the least-squares method. Two heat transfer models appeared to fit the data well and were equivalent: conductive heat transfer alone, and convective plus radiative heat transfers. * CONCLUSIONS: The increase in the temperature difference between the spadix and ambient air appears to be physically constrained and corresponds to the value of a thermal model of heat conduction in an insulated cylinder with an internal heat source. In the models, a heat metabolic rate of 29.5 mW g(-1) was used, which was an acceptable value for an overall metabolic heat rate in aroid inflorescences. PMID- 15883131 TI - Suppressed expression of the apoplastic ascorbate oxidase gene increases salt tolerance in tobacco and Arabidopsis plants. AB - Transgenic tobacco plants expressing the ascorbate oxidase (AAO) gene in sense and antisense orientations, and an Arabidopsis mutant in which the T-DNA was inserted into a putative AAO gene, were used to examine the potential roles of AAO for salt-stress tolerance in plants. AAO activities in the transgenic tobacco plants expressing the gene in sense and antisense orientations were, respectively, about 16-fold and 0.2-fold of those in the wild type. Under normal growth conditions, no significant differences in phenotypes were observed, except for a delay in flowering time in the antisense plants. However, at high salinity, the percentage germination, photosynthetic activity, and seed yields were higher in antisense plants, with progressively lower levels in the wild type and the sense plants. The redox state of apoplastic ascorbate in sense plants was very low even under normal growth conditions. Upon salt stress, the redox state of symplastic and apoplastic ascorbate decreased among the three types of plants, but was lowest in the sense plants. The hydrogen peroxide contents in the symplastic and apoplastic spaces were higher in sense plants, progressively lower in the wild type, followed by the antisense plants. The Arabidopsis T-DNA inserted mutant exhibited very low ascorbate oxidase activity, and its phenotype was similar to that of antisense tobacco plants. These results suggest that the suppressed expression of apoplastic AAO under salt-stress conditions leads to a relatively low level of hydrogen peroxide accumulation and a high redox state of symplastic and apoplastic ascorbate which, in turn, permits a higher seed yield. PMID- 15883132 TI - One year clinical follow up of paclitaxel eluting stents for acute myocardial infarction compared with sirolimus eluting stents. AB - OBJECTIVE: To compare clinical outcome of paclitaxel eluting stents (PES) versus sirolimus eluting stents (SES) for the treatment of acute ST elevation myocardial infarction. DESIGN AND PATIENTS: The first 136 consecutive patients treated exclusively with PES in the setting of primary percutaneous coronary intervention for acute myocardial infarction in this single centre registry were prospectively clinically assessed at 30 days and one year. They were compared with 186 consecutive patients treated exclusively with SES in the preceding period. SETTING: Academic tertiary referral centre. RESULTS: At 30 days, the rate of all cause mortality and reinfarction was similar between groups (6.5% v 6.6% for SES and PES, respectively, p = 1.0). A significant difference in target vessel revascularisation (TVR) was seen in favour of SES (1.1% v 5.1% for PES, p = 0.04). This was driven by stent thrombosis (n = 4), especially in the bifurcation stenting (n = 2). At one year, no significant differences were seen between groups, with no late thrombosis and 1.5% in-stent restenosis (needing TVR) in PES versus no reinterventions in SES (p = 0.2). One year survival free of major adverse cardiac events (MACE) was 90.2% for SES and 85% for PES (p = 0.16). CONCLUSIONS: No significant differences were seen in MACE-free survival at one year between SES and PES for the treatment of acute myocardial infarction with very low rates of reintervention for restenosis. Bifurcation stenting in acute myocardial infarction should, if possible, be avoided because of the increased risk of stent thrombosis. PMID- 15883133 TI - Evaluation of survival and ischaemic and thromboembolic event rates in patients with non-valvar atrial fibrillation in the general population when treated and untreated with warfarin. AB - OBJECTIVE: To compare survival and adverse outcome of patients with non-valvar atrial fibrillation (NVAF) treated with or without warfarin. DESIGN: Record linkage method to identify patients with a previous hospital diagnosis of atrial fibrillation and to link these patients to international normalised ratio (INR) test results and mortality data. SETTING: Cardiff and the Vale of Glamorgan, Wales. MAIN OUTCOME MEASURES: Mortality, specifically from ischaemic and thromboembolic events. RESULTS: 6108 patients were identified with NVAF, of whom 36.4% received warfarin. Mean survival in the warfarin and non-warfarin groups was 52.0 months and 38.2 months, respectively (p < 0.001), and 14.4 months (p < 0.001) after adjustment for confounding factors. Warfarin treated patients in the upper quartile of INR control had significantly longer survival (57.5 months) than did those in the lowest quartile of control (38.1 months, p < 0.001). The risk of stroke in the warfarin group when treated was lower than that in the non warfarin group (relative rate (RR) 0.74, p < 0.001). The risk of death from ischaemic stroke was lower in the warfarin group (RR 0.43, p < 0.001). The risk of all ischaemic and embolic events in the warfarin group was lower when they were taking warfarin (RR 0.74, p < 0.001). The risk of bleeding in the warfarin group when treated was greater (RR 1.78, p = 0.001). CONCLUSIONS: Patients with NVAF within the recommended target INR range of 2.0-3.0 survive longer and have reduced morbidity. Probably too few people are anticoagulated with warfarin in NVAF. PMID- 15883134 TI - Shortening the stent length reduces restenosis with bare metal stents: matched pair comparison of short stenting and conventional stenting. AB - OBJECTIVE: To investigate the effect of reducing stent length on the rate of target lesion restenosis. DESIGN: In a prospective investigation, acute and long term results of a short stenting procedure were analysed by quantitative angiography and compared with results of a conventional stenting procedure selected according to a matched pairs analysis. PATIENTS: Short stents were implanted in 400 consecutive patients with 464 lesions and conventional stents in 430 patients. Demographic and lesion characteristics were comparable between groups. INTERVENTIONS: In short stenting, the shortest stent length to cover only segments with > 30% reduction in vessel diameter was used. In conventional stenting, full coverage of a stenotic vessel segment was intended. MAIN OUTCOME MEASURES: The mean stent lengths of the short stent group (9.8 (4) mm) and the conventional stent group (16.3 (7) mm) differed significantly (p < 0.0001); all other procedural and angiographic parameters were the same. Procedural success was similar for both groups. Control angiography after six months was conducted in 92% of patients. RESULTS: Short stenting resulted in both less restenosis (68 of 431 (15.8%)) than conventional stenting (93 of 381 (24.4%), p = 0.007) and less late lumen loss (0.6 (0.6) mm v 0.75 (0.5) mm, p = 0.0001). Residual stenosis (< 45%) in adjacent vessel segments after short stenting did not affect the restenosis rate. Only the implantation of a < or = 9 mm stent predicted the absence of restenosis in a multivariate analysis. CONCLUSION: Shortening the length of bare metal stents reduces the restenosis rate as compared with conventional stenting. PMID- 15883135 TI - Circumstances of out of hospital cardiac arrest in patients with ischaemic heart disease. AB - OBJECTIVES: To discover the circumstances of out of hospital cardiac death irrespective of resuscitation attempts. DESIGN: Prospective community study over the two years 1994 and 1995. SETTING: The health districts of Brighton, South Glamorgan, and York, UK. SUBJECTS: 1290 victims of sudden death or cardiac arrest caused by coronary heart disease who were under 76 years of age. INTERVENTIONS: Basic and advanced life support for witnessed cardiac arrests. MAIN OUTCOME MEASURES: Survival to reach hospital and for 30 days after the arrest. RESULTS: 35 (35%) of 101 patients (mean age 64) whose arrest was witnessed by a doctor or paramedic survived for 30 days compared with 9 of 464 (2%) whose arrest was witnessed by a relative or bystander at home (mean age 66) and 15 of 200 (8%) whose arrest was witnessed in a public place (mean age 61). None of the 525 victims of an unwitnessed arrest survived but the majority of those whose arrest was witnessed had complained of new symptoms before the arrest. Victims who were given basic life support by relatives or bystanders had better survival (14 of 183 (8%)) than those who were not (10 of 481 (2%), p < 0.001). Of the 20% of arrests that occurred in public places, few were in places where public access defibrillators would now be available. CONCLUSIONS: The burden of out of hospital cardiac arrest is mainly in the home but most victims have premonitory symptoms. Public education to seek help urgently for new or prolonged chest pain seems the most promising method to address the problem. PMID- 15883136 TI - Can poorly performing doctors blame their assessment tools? PMID- 15883137 TI - Use of SPRAT for peer review of paediatricians in training. AB - OBJECTIVE: To determine whether a multisource feedback questionnaire, SPRAT (Sheffield peer review assessment tool), is a feasible and reliable assessment method to inform the record of in-training assessment for paediatric senior house officers and specialist registrars. DESIGN: Trainees' clinical performance was evaluated using SPRAT sent to clinical colleagues of their choosing. Responses were analysed to determine variables that affected ratings and their measurement characteristics. SETTING: Three tertiary hospitals and five secondary hospitals across a UK deanery. PARTICIPANTS: 112 paediatric senior house officers and middle grades. MAIN OUTCOME MEASURES: 95% confidence intervals for mean ratings; linear and hierarchical regression to explore potential biasing factors; time needed for the process per doctor. RESULTS: 20 middle grades and 92 senior house officers were assessed using SPRAT to inform their record of in-training assessment; 921/1120 (82%) of their proposed raters completed a SPRAT form. As a group, specialist registrars (mean 5.22, SD 0.34) scored significantly higher (t = - 4.765) than did senior house officers (mean 4.81, SD 0.35) (P < 0.001). The grade of the doctor accounted for 7.6% of the variation in the mean ratings. The hierarchical regression showed that only 3.4% of the variation in the means could be additionally attributed to three main factors (occupation of rater, length of working relationship, and environment in which the relationship took place) when the doctor's grade was controlled for (significant F change < 0.001). 93 (83%) of the doctors in this study would have needed only four raters to achieve a reliable score if the intent was to determine if they were satisfactory. The mean time taken to complete the questionnaire by a rater was six minutes. Just over an hour of administrative time is needed for each doctor. CONCLUSIONS: SPRAT seems to be a valid way of assessing large numbers of doctors to support quality assurance procedures for training programmes. The feedback from SPRAT can also be used to inform personal development planning and focus quality improvements. PMID- 15883138 TI - Barriers to concordance with antidiabetic drugs--cultural differences or human nature? PMID- 15883139 TI - Perceptions and experiences of taking oral hypoglycaemic agents among people of Pakistani and Indian origin: qualitative study. AB - OBJECTIVE: To explore British Pakistani and British Indian patients' perceptions and experiences of taking oral hypoglycaemic agents (OHAs). DESIGN: Observational cross sectional study using in-depth interviews in English or Punjabi. SETTING AND PARTICIPANTS: 32 patients of Pakistani and Indian origin with type 2 diabetes, recruited from primary care and community sources in Edinburgh, Scotland. RESULTS: Respondents reported complex and ambivalent views about OHAs, which reflected their ambivalent attitudes towards Western drugs in general. Respondents considered OHAs to be an important part of the diabetic regimen because they perceived British healthcare professionals to be competent and trustworthy prescribers, and they considered the medicines available in Britain to be superior to those on the Indian subcontinent. Despite this, some respondents made deliberate efforts to reduce their tablet intake without being advised to do so. Reasons for this included perceptions that drugs worked by providing relief of symptoms and concerns that OHAs could be detrimental to health if taken for long periods, in conjunction with other drugs, or without traditional foods. CONCLUSIONS: British Pakistani and Indian patients' perceptions of their OHAs may partly derive from popular ideas about drugs on the Indian subcontinent. Cultural factors need to be understood and taken into consideration to ensure that these patients are given appropriate advice and to avoid unnecessary changes to prescriptions. PMID- 15883140 TI - Definition, diagnosis, and forensic implications of postconcussional syndrome. AB - Injuries from blows to the head often are manifested only as subjective complaints. Postconcussional syndrome thus can be feigned for financial or psychological gain. The authors review the pathology of brain trauma, symptoms of postconcussional syndrome, and criteria for diagnosis. In addition to somatic deficits, psychological and cognitive problems are common. The likelihood and severity of postconcussional syndrome are greater for women. Malingering may be suspected in cases involving litigation, and tests to detect it are available. Treatment for postconcussional syndrome depends on the specific symptoms. Pharmacotherapies may be helpful, but care should be used in prescribing drugs that could produce deleterious CNS effects. PMID- 15883141 TI - Biomedical and psychosocial determinants of intrusive recollections in breast cancer survivors. AB - The authors' aim was to identify the determinants of intrusive recollections related to receiving a cancer diagnosis in women after cancer treatment. A consecutive series of breast cancer patients at 3-15 months after surgery, consisting of subjects with (N=63) and without (N=92) intrusive recollections was examined. Logistic regression analysis revealed that neuroticism, precancer intrusive recollections, and the number of cancer patients in relatives by marriage were final significant determinants, and receiving radiotherapy was an associated factor. These results suggest that biopsychosocial factors contribute to intrusive recollections and have clinical implications for the use of these characteristics in screening and early intervention. PMID- 15883142 TI - Determinants of health-related quality of life in coronary artery disease patients: a prospective study generating a structural equation model. AB - The authors used structural equation modeling to test a conceptual model of HRQL in coronary artery disease. The model, which included biomedical factors and individual and environmental characteristics, was tested in a multicenter group of 465 patients at three timepoints (baseline evaluation of chest pain and 1- and 3-month follow-ups). A satisfactory fit was obtained for the model over time. Depression and anxiety symptoms exerted the most significant influence on HRQL. HRQL and the mediating factors were found to be distinct phenomena. The authors concluded that mediating factors, especially depression and anxiety symptoms, should be taken into consideration in clinical routine if HRQL is regarded as a clinical outcome. PMID- 15883143 TI - Can collaborative care address the needs of low-income Latinas with comorbid depression and cancer? Results from a randomized pilot study. AB - In a pilot study, 55 low-income Latina patients with breast or cervical cancer and comorbid depression were randomly assigned to receive collaborative care as part of the Multifaceted Oncology Depression Program or usual care. Relative to patients in the usual care condition, patients receiving collaborative care were more likely to show>or=50% improvement in depressive symptoms as measured by the Personal Health Questionnaire (OR=4.51, 95% CI=1.07-18.93). Patients in the collaborative care program were also more likely to show improvement in emotional well-being (increase of 2.15) as measured by the Functional Assessment of Cancer Therapy Scale than were those receiving usual care (decrease of 0.50) (group difference=2.65, 95% CI: 0.18-5.12). Despite health system, provider, and patient barriers to care, these initial results suggest that patients in public sector oncology clinics can benefit from onsite depression treatment. PMID- 15883144 TI - Relationship of tumor burden and patients' minimization of distress in facing surgery for gastric cancer. AB - Patients with gastric cancer considered amenable to a curative surgical procedure on the basis of a conventional preoperative workup (N=98) answered health-related quality-of-life questionnaires and completed a psychiatrist's interview before the intervention. All patients were blinded to the type of surgical procedure that would ultimately be performed; 64 patients eventually received a potentially curative resection, and 34 had palliative surgery. The curative resection group reported significantly more hopelessness before the operation than the palliative surgery group. The latter group had more eating dysfunction but minimized their distress to a greater degree than those with a more limited tumor burden. Eating dysfunction, hopelessness, and the tendency to minimize distress were independent predictors of prognosis. The findings highlight the need for careful medical interviewing to enhance patient-physician communication about symptoms and signs of malignant diseases. PMID- 15883145 TI - Major depression and the use of electroconvulsive therapy (ECT) in lung transplant recipients. AB - The purpose of this study is to describe the potential risks and benefits of electroconvulsive therapy (ECT) for treatment of depression in lung transplant recipients. The authors performed a record review of depressed patients who underwent lung transplantation at Johns Hopkins Hospital and evaluated their treatment, including ECT. In 9 years, 131 lung transplants were performed, and four patients had been diagnosed with major depression. Of those, two were candidates for ECT, and one received it. This patient's depression did abate with ECT. ECT, an effective treatment for depression, remains a treatment method of choice for depression in the posttransplant population. PMID- 15883146 TI - Dissolution of common psychiatric medications in a Roux-en-Y gastric bypass model. AB - Large numbers of Roux-en-Y gastric bypass (RYGB) surgery patients have psychiatric illnesses that are in part treated with medication preoperatively, but there are no objective data to guide psychiatric drug dosing postoperatively. An in vitro drug dissolution model was developed to approximate the gastrointestinal environment of the preoperative (control) and post-RYGB states. Medication tablets were placed in the two environments, and the median calculated weights of the dissolved portions were compared. Ten of 22 psychiatric medication preparations had significantly less dissolution and two had significantly greater dissolution in the post-RYGB environment, compared with the control environment. The results suggest a need for an in vivo study of serum drug levels after RYGB surgery in patients taking psychiatric medications. Differences in the pharmacokinetics of the postoperative RYGB patient may necessitate adjustments in dosing. PMID- 15883147 TI - Risperidone treatment of three seriously medically ill children with secondary mood disorders. PMID- 15883148 TI - Acute mania in the setting of severe hypothyroidism. AB - Although the associations between depression and hypothyroidism and between mania and hyperthyroidism are well described, mania in the setting of hypothyroidism is unusual. The authors present the case of a patient whose acute mania appears to have been precipitated by hypothyroidism secondary to postpartum thyroiditis. This case underscores the importance of thyroid screening in patients with mood and psychotic disorders, including patients who lack the classical psychiatric features of thyroid dysfunction. Further investigation is required on the nature of the relationship between thyroid function and bipolar disorder and any implications it may have for the diagnosis and treatment of this illness. PMID- 15883149 TI - The dosing of atypical antipsychotics. AB - Drug-drug interactions or genetic variability may require using doses different from those recommended for atypical antipsychotics. Dosage alterations of olanzapine and clozapine, dependent on cytochrome P450 1A2 (CYP1A2) for clearance, and quetiapine, dependent on cytochrome P450 3A (CYP3A), may be necessary when used with other drugs that inhibit or induce their metabolic enzymes. Smoking cessation can significantly increase clozapine, and perhaps olanzapine, levels. Ziprasidone pharmacokinetic drug-drug interactions are not likely to be important. Genetic variations of cytochrome P450 2D6 (CYP2D6) and drug-drug interactions causing inhibition (CYP2D6 and/or CYP3A) or induction (CYP3A) may be important for risperidone, and perhaps for aripiprazole, dosing. Adding inhibitors may cause side effects more easily in drugs with a narrow therapeutic window, such as clozapine or risperidone, than in those with a wide therapeutic window, such as olanzapine or aripiprazole. Adding inducers may be associated with a gradual development of lost efficacy. PMID- 15883150 TI - Serotonin syndrome associated with linezolid treatment after discontinuation of fluoxetine. PMID- 15883152 TI - What makes a successful consultant in psychosomatic medicine? PMID- 15883151 TI - Haloperidol-induced ileus. PMID- 15883153 TI - Properties of WNK1 and implications for other family members. AB - WNKs are large serine/threonine protein kinases structurally distinct from all other members of the protein kinase superfamily. Of the four human WNK family members, WNK1 and WNK4 have been linked to a hereditary form of hypertension, pseudohypoaldosteronism type II. We characterized the biochemical properties and regulation of WNK1 that may contribute to its physiological activities and abnormal function in disease. We showed that WNK1 is activated by hypertonic stress in kidney epithelial cells and in breast and colon cancer cell lines. In addition, hypotonic stress also led to a modest increase in WNK1 activity. Gel filtration suggested that WNK1 exists as a tetramer, and yeast two-hybrid data showed that the N terminus of WNK1 (residues 1-222) interacts with residues 481 660, which includes the WNK1 autoinhibitory domain and a C-terminal coiled-coil domain. Although cell biological studies have suggested a functional interaction between WNK1 and WNK4, we found no evidence of stable interactions between these kinases. However, WNK1 phosphorylated both WNK4 and WNK2. In addition, the WNK1 autoinhibitory domain inhibited the catalytic activity of these WNKs. These findings suggest potential mechanisms for interconnected regulation of WNK family members. PMID- 15883154 TI - Rapid transbilayer movement of ceramides in phospholipid vesicles and in human erythrocytes. AB - The transbilayer diffusion of unlabeled ceramides with different acyl chains (C6 Cer, C10-Cer, and C16-Cer) was investigated in giant unilamellar vesicles (GUVs) and in human erythrocytes. Incorporation of a very small percentage of ceramides (approximately 0.1% of total lipids) to the external leaflet of egg phosphatidylcholine GUVs suffices to trigger a shape change from prolate to pear shape vesicle. By observing the reversibility of this shape change the transmembrane diffusion of lipids was inferred. We found a half-time for unlabeled ceramide flip-flop below 1 min at 37 degrees C. The rapid diffusion of ceramides in a phosphatidylcholine bilayer was confirmed by flip-flop experiments with a spin-labeled ceramide analogue incorporated into large unilamellar vesicles. Shape change experiments were also carried out with human erythrocytes to determine the trans-membrane diffusion of unlabeled ceramides into a biological membrane. Addition of exogenous ceramides to the external leaflet of human erythrocytes did not trigger echinocyte formation immediately as one would anticipate from an asymmetrical accumulation of new amphiphiles in the outer leaflet but only after approximately 15 min of incubation at 20 degrees C in the presence of an excess of ceramide. We interpret these data as being indicative of a rapid ceramide equilibration between both erythrocyte leaflets as indicated also by electron spin resonance spectroscopy with a spin-labeled ceramide. The late appearance of echinocytes could reveal a progressive trapping of a fraction of the ceramide molecules in the outer erythrocytes leaflet. Thus, we cannot exclude the trapping of ceramides into plasma membrane domains. PMID- 15883155 TI - Glycodelin-S in human seminal plasma reduces cholesterol efflux and inhibits capacitation of spermatozoa. AB - Tight control of sperm capacitation is important for successful fertilization. Glycodelin-S is one of the most abundant glycoproteins in the human seminal plasma. However, its function is unclear. We investigated the role of glycodelin S on capacitation of human spermatozoa. Binding kinetics experiments demonstrated the presence of two saturable and reversible binding sites of glycodelin-S on human spermatozoa. Differently glycosylated other isoforms of glycodelin, glycodelin-A and -F, did not compete with glycodelin-S for these binding sites, suggesting that the glycodelin-S binding sites are different from those of the other isoforms. Indirect immunofluorescent staining revealed specific binding of glycodelin-S around the sperm head. This immunoreactivity was greatly reduced in spermatozoa that had migrated through the cervical mucus surrogates. Glycodelin-S at physiological concentrations significantly reduced the bovine serum albumin and cyclodextrin-induced cholesterol efflux and down-regulated the adenylyl cyclase/protein kinase A/tyrosine kinase signaling pathway, resulting in suppression of capacitation. Deglycosylation abolished glycodelin-S binding and the effect of glycodelin-S on bovine serum albumin-induced capacitation. This indicates that the carbohydrate moiety of glycodelin-S is critical for the function of the molecule. It is concluded that glycodelin-S in seminal plasma maintains the uncapacitated state of human spermatozoa. PMID- 15883156 TI - Dictyostelium discoideum expresses a malaria chloroquine resistance mechanism upon transfection with mutant, but not wild-type, Plasmodium falciparum transporter PfCRT. AB - Chloroquine resistance in Plasmodium falciparum malaria results from mutations in PfCRT, a member of a unique family of transporters present in apicomplexan parasites and Dictyostelium discoideum. Mechanisms that have been proposed to explain chloroquine resistance are difficult to evaluate within malaria parasites. Here we report on the targeted expression of wild-type and mutant forms of PfCRT to acidic vesicles in D. discoideum. We show that wild-type PfCRT has minimal effect on the accumulation of chloroquine by D. discoideum, whereas forms of PfCRT carrying a key charge-loss mutation of lysine 76 (e.g. K76T) enable D. discoideum to expel chloroquine. As in P. falciparum, the chloroquine resistance phenotype conferred on transformed D. discoideum can be reversed by the channel-blocking agent verapamil. Although intravesicular pH levels in D. discoideum show small acidic changes with the expression of different forms of PfCRT, these changes would tend to promote intravesicular trapping of chloroquine (a weak base) and do not account for reduced drug accumulation in transformed D. discoideum. Our results instead support outward-directed chloroquine efflux for the mechanism of chloroquine resistance by mutant PfCRT. This mechanism shows structural specificity as D. discoideum transformants that expel chloroquine do not expel piperaquine, a bisquinoline analog of chloroquine used frequently against chloroquine-resistant parasites in Southeast Asia. PfCRT, nevertheless, may have some ability to act on quinine and quinidine. Transformed D. discoideum will be useful for further studies of the chloroquine resistance mechanism and may assist in the development and evaluation of new antimalarial drugs. PMID- 15883157 TI - Functional characterization of novel alternatively spliced ClC-2 chloride channel variants in the heart. AB - A novel volume-regulated hyperpolarization-activated chloride inward rectifier channel (Cl.ir) was identified in mammalian heart. To investigate whether ClC-2 is the gene encoding Cl.ir channels in heart, ClC-2 cDNAs cloned from rat (rClC 2) and guinea pig (gpClC-2) hearts were functionally characterized. When expressed in NIH/3T3 cells, full-length rClC-2 yielded inwardly rectifying whole cell currents with very slow activation kinetics (time constants > 1.7 s) upon hyperpolarization under hypotonic condition. The single-channel rClC-2 currents had a unitary slope conductance of 3.9 +/- 0.2 picosiemens. A novel variant with an in-frame deletion at the beginning of exon 15 that leads to a deletion of 45 bp (corresponding to 15 amino acids in alpha-helices O and P, rClC-2(Delta509 523)) was identified in rat heart. The relative transcriptional expression levels of full-length rClC-2 and rClC-2(Delta509-523) in rat heart were 0.018 +/- 0.003 and 0.028 +/- 0.006 arbitrary units, respectively, relative to glyceraldehyde-3 phosphate dehydrogenase (n = 5, p = nonsignificant). A similar partial exon 15 skipping with a deletion of 105 bp (35 amino acids in alpha-helices O-Q, gpClC 2(Delta509-543)) was also identified in guinea pig heart. Expression of both rClC 2(Delta509-523) and gpClC-2(Delta509-543) resulted in functional channels with phenotypic activation kinetics and many properties identical to those of endogenous Cl.ir channels in native rat and guinea pig cardiac myocytes, respectively. Intracellular dialysis of anti-ClC-2 antibody inhibited expressed ClC-2 channels and endogenous Cl.ir currents in native rat and guinea pig cardiac myocytes. These results demonstrate that novel deletion variants of ClC-2 due to partial exon 15 skipping may be expressed normally in heart and contribute to the formation of endogenous Cl.ir channels in native cardiac cells. PMID- 15883158 TI - Bone morphogenetic protein (BMP) type II receptor deletion reveals BMP ligand specific gain of signaling in pulmonary artery smooth muscle cells. AB - Bone morphogenetic protein (BMP) ligands signal by binding the BMP type II receptor (BMPR2) or the activin type II receptors (ActRIIa and ActRIIb) in conjunction with type I receptors to activate SMADs 1, 5, and 8, as well as members of the mitogen-activated protein kinase family. Loss-of-function mutations in Bmpr2 have been implicated in tumorigenesis and in the etiology of primary pulmonary hypertension. Because several different type II receptors are known to recognize BMP ligands, the specific contribution of BMPR2 to BMP signaling is not defined. Here we report that the ablation of Bmpr2 in pulmonary artery smooth muscle cells, using an ex vivo conditional knock-out (Cre-lox) approach, as well as small interfering RNA specific for Bmpr2, does not abolish BMP signaling. Disruption of Bmpr2 leads to diminished signaling by BMP2 and BMP4 and augmented signaling by BMP6 and BMP7. Using small interfering RNAs to inhibit the expression of other BMP receptors, we found that wild-type cells transduce BMP signals via BMPR2, whereas BMPR2-deficient cells transduce BMP signals via ActRIIa in conjunction with a set of type I receptors distinct from those utilized by BMPR2. These findings suggest that disruption of Bmpr2 leads to the net gain of signaling by some, but not all, BMP ligands via the activation of ActRIIa. PMID- 15883159 TI - An obligatory intermediate in the folding pathway of cytochrome c552 from Hydrogenobacter thermophilus. AB - The folding mechanism of many proteins involves the population of partially organized structures en route to the native state. Identification and characterization of these intermediates is particularly difficult, as they are often only transiently populated and may play different mechanistic roles, being either on-pathway productive species or off-pathway kinetic traps. Following different spectroscopic probes, and employing state-of-the-art kinetic analysis, we present evidence that the folding mechanism of the thermostable cytochrome c552 from Hydrogenobacter thermophilus does involve the presence of an elusive, yet compact, on-pathway intermediate. Characterization of the folding mechanism of this cytochrome c is particularly interesting for the purpose of comparative folding studies, because H. thermophilus cytochrome c552 shares high sequence identity and structural homology with its homologue from the mesophilic bacterium Pseudomonas aeruginosa cytochrome c551, which refolds through a broad energy barrier without the accumulation of intermediates. Analysis of the folding kinetics and correlation with the three-dimensional structure add new evidence for the validity of a consensus folding mechanism in the cytochrome c family. PMID- 15883160 TI - Ketone bodies stimulate chaperone-mediated autophagy. AB - Chaperone-mediated autophagy (CMA) is a selective lysosomal protein degradative process that is activated in higher organisms under conditions of prolonged starvation and in cell culture by the removal of serum. Ketone bodies are comprised of three compounds (beta-hydroxybutyrate, acetoacetate, and acetone) that circulate during starvation, especially during prolonged starvation. Here we have investigated the hypothesis that ketone bodies induce CMA. We found that physiological concentrations of beta-hydroxybutyrate (BOH) induced proteolysis in cells maintained in media with serum and without serum; however, acetoacetate only induced proteolysis in cells maintained in media with serum. Lysosomes isolated from BOH-treated cells displayed an increased ability to degrade both glyceraldehyde-3-phosphate dehydrogenase and ribonuclease A, substrates for CMA. Isolated lysosomes from cells maintained in media without serum also demonstrated an increased ability to degrade glyceraldehyde-3-phosphate dehydrogenase and ribonuclease A when the reaction was supplemented with BOH. Such treatment did not affect the levels of lysosome-associated membrane protein 2a or lysosomal heat shock cognate protein of 70 kDa, two rate-limiting proteins in CMA. However, pretreatment of glyceraldehyde-3-phosphate and ribonuclease A with BOH increased their rate of degradation by isolated lysosomes. Lysosomes pretreated with BOH showed no increase in proteolysis, suggesting that BOH acts on the substrates to increase their rates of proteolysis. Using OxyBlot analysis to detect carbonyl formation on proteins, one common marker of protein oxidation, we showed that treatment of substrates with BOH increased their oxidation. Neither glycerol, another compound that increases in circulation during prolonged starvation, nor butanol or butanone, compounds closely related to BOH, had an effect on CMA. The induction of CMA by ketone bodies may provide an important physiological mechanism for the activation of CMA during prolonged starvation. PMID- 15883161 TI - Alpha-kinase 1, a new component in apical protein transport. AB - A key aspect in the structure of epithelial cells is the maintenance of a polarized organization based on a highly specific sorting machinery for cargo destined for the apical or the basolateral membrane domain at the exit site of the trans-Golgi network. We could recently identify two distinct post-trans-Golgi network vesicle populations that travel along separate routes to the plasma membrane, a lipid raft-dependent and a lipid raft-independent pathway. A new component of raft-carrying apical vesicles is alpha-kinase 1 (ALPK1), which was identified in immunoisolated vesicles carrying raft-associated sucrase-isomaltase (SI). This kinase was absent from vesicles carrying raft-non-associated lactase phlorizin hydrolase. The expression of ALPK1 increases by the time of epithelial cell differentiation, whereas the intracellular localization of ALPK1 on apical transport vesicles was confirmed by confocal analysis. A phosphorylation assay on isolated SI-carrying vesicles revealed the phosphorylation of a protein band of about 105 kDa, which could be identified as the motor protein myosin I. Finally, a specific reduction of ALPK1-expression by RNA interference results in a significant decrease in the apical delivery of SI. Taken together, our data suggest that the phosphorylation of myosin I by ALPK1 is an essential process in the apical trafficking of raft-associated SI. PMID- 15883162 TI - Glutathione redox state regulates mitochondrial reactive oxygen production. AB - Oxidative stress induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; dioxin) is poorly understood. Following one dose of TCDD (5 microg/kg body weight), mitochondrial succinate-dependent production of superoxide and H2O2 in mouse liver doubled at 7-28 days, then subsided by day 56; concomitantly, levels of GSH and GSSG increased in both cytosol and mitochondria. Cytosol displayed a typical oxidative stress response, consisting of diminished GSH relative to GSSG, decreased potential to reduce protein-SSG mixed disulfide bonds (type 1 thiol redox switch) or protein-SS-protein disulfide bonds (type 2 thiol redox switch), and a +10 mV change in GSSG/2GSH reduction potential. In contrast, mitochondria showed a rise in reduction state, consisting of increased GSH relative to GSSG, increases in type 1 and type 2 thiol redox switches, and a -25 mV change in GSSG/2GSH reduction potential. Comparing Ahr(-/-) knock-out and wild-type mice, we found that TCDD-induced thiol changes in both cytosol and mitochondria were dependent on the aromatic hydrocarbon receptor (AHR). GSH was rapidly taken up by mitochondria and stimulated succinate-dependent H2O2 production. A linear dependence of H2O2 production on the reduction potential for GSSG/2GSH exists between -150 and -300 mV. The TCDD-stimulated increase in succinate-dependent and thiol-stimulated production of reactive oxygen paralleled a four-fold increase in formamidopyrimidine DNA N-glycosylase (FPG)-sensitive cleavage sites in mitochondrial DNA, compared with a two-fold increase in nuclear DNA. These results suggest that TCDD produces an AHR-dependent oxidative stress in mitochondria, with concomitant mitochondrial DNA damage mediated, at least in part, by an increase in the mitochondrial thiol reduction state. PMID- 15883163 TI - Characterization of mitochondrial and extra-mitochondrial oxygen consuming reactions in human hematopoietic stem cells. Novel evidence of the occurrence of NAD(P)H oxidase activity. AB - This study was aimed to characterize the mitochondrial and extra-mitochondrial oxygen consuming reactions in human CD34+ hematopoietic stem cells. Cell samples were collected by apheresis following pre-conditioning by granulocyte colony stimulating factor and isolated by anti-CD34 positive immunoselection. Polarographic analysis of the CN-sensitive endogenous cell respiration revealed a low mitochondrial oxygen consumption rate. Differential absorbance spectrometry on whole cell lysate and two-dimensional blue native-PAGE analysis of mitoplast proteins confirmed a low amount of mitochondrial respiratory chain complexes thus qualifying the hematopoietic stem cell as a poor oxidative phosphorylating cell type. Confocal microscopy imaging showed, however, that the intracellular content of mitochondria was not homogeneously distributed in the CD34+ hematopoietic stem cell sample displaying a clear inverse correlation of their density with the expression of the CD34 commitment marker. About half of the endogenous oxygen consumption was extra-mitochondrial and completely inhibitable by enzymatic scavengers of reactive oxygen species and by diphenylene iodinium. By spectral analysis, flow cytometry, reverse transcriptase-PCR, immunocytochemistry, and immunoprecipitation it was shown that the extra-mitochondrial oxygen consumption was contributed by the NOX2 and NOX4 isoforms of the O2-*. producer plasma membrane NAD(P)H oxidase with low constitutive activity. A model is proposed suggesting for the NAD(P)H oxidase a role of O2 sensor and/or ROS source serving as redox messengers in the activation of intracellular signaling pathways leading (or contributing) to mitochondriogenesis, cell survival, and differentiation in hematopoietic stem cells. PMID- 15883164 TI - Interferon alpha activates NF-kappaB in JAK1-deficient cells through a TYK2 dependent pathway. AB - In addition to activating members of the STAT transcription factor family, interferon alpha/beta (IFNalpha/beta) activates the NF-kappaB transcription factor. To determine the role of the Janus tyrosine kinase (JAK)-STAT pathway in NF-kappaB activation by IFN, we examined NF-kappaB activation in JAK1-deficient mutant human fibrosarcoma cells. In wild-type fibrosarcoma cells (2fTGH), IFN activates STAT1, STAT2, and STAT3, as well as NF-kappaB complexes comprised of p50 and p65. In contrast, in JAK1-deficient cells, IFN induces NF-kappaB activation and NF-kappaB dependent gene transcription but does not activate these STAT proteins and has no effect on STAT-dependent gene transcription. Expression of a catalytically inactive TYK2 tyrosine kinase in JAK1-deficient cells, as well as in the highly IFN-sensitive Daudi lymphoblastoid cell line, abrogates NF kappaB activation by IFN. Moreover, IFN does not promote NF-kappaB activation in TYK2-deficient mutant fibrosarcoma cells. Our results demonstrate a dichotomy between the classical JAK-STAT pathway and the NF-kappaB signaling pathway. In the IFN signaling pathway leading to STAT activation, both JAK1 and TYK2 are essential, whereas NF-kappaB activation requires only TYK2. PMID- 15883165 TI - Sphingosine activates protein kinase A type II by a novel cAMP-independent mechanism. AB - Protein kinase A (PKA) has long been recognized as playing a major role in many regulatory processes in cells through its activation by the ubiquitous second messenger cAMP. We show here a novel mode of activation of PKA type II that is independent of cAMP and is, instead, dependent on sphingosine. PKA type II is specifically activated by sphingosine and its analog, dimethylsphingosine, but not by sphingosine-1-phosphate or other lipids. Like cAMP, sphingosine activates PKA holoenzyme but not the catalytic subunit alone, suggesting that the activation is mediated by the regulatory subunits. However, sphingosine-activated PKA, but not cAMP-activated PKA, is inhibited by phosphatidylserine, suggesting a distinct mechanism of activation. Furthermore, unlike cAMP, sphingosine does not induce the dissociation of PKA holoenzyme into catalytic and regulatory subunits. Modulation of sphingosine levels in vivo results in alteration in basal membrane associated PKA activity consistent with a direct effect of membrane sphingosine on PKA type II. Importantly, sphingosine-dependent but not cAMP-dependent activation of PKA specifically phosphorylates Ser58 of the multifunctional adapter protein 14-3-3zeta, promoting the conversion of dimeric 14-3-3 to a monomeric state, thus potentially modulating several biological functions. These results define a new mode of PKA activation that is sphingosine-dependent and mechanistically different from the classical cAMP-dependent activation of PKA. Furthermore, they suggest that stimuli that induce sphingosine accumulation and modulate phospholipid content at the cell membrane have the potential to activate PKA, thereby inducing the phosphorylation of distinct substrates and biological activities. PMID- 15883166 TI - Multiphoton imaging of renal tissues in vitro. AB - The highly inhomogeneous and light-scattering structure of living renal tissue makes the application of conventional imaging techniques more difficult compared with other parenchymal organs. On the other hand, key physiological processes of the kidney, such as regulation of glomerular filtration, hemodynamics, concentration, and dilution, involve complex interactions between multiple cell types and otherwise inaccessible structures that necessitate visual approaches. An ideal solution is multiphoton excitation fluorescence microscopy, a state-of the-art imaging technique superior for deep optical sectioning of living tissue samples. Here, we review the basics and advantages of multiphoton microscopy and provide examples for its application in renal physiology using dissected cortical and medullary tissues in vitro. In combination with microperfusion techniques, the major functions of the juxtaglomerular apparatus, tubuloglomerular feedback and renin release, can be studied with high spatial and temporal resolution. Salt dependent changes in macula densa cell volume, vasoconstriction of the afferent arteriole, and activity of an intraglomerular precapillary sphincter composed of renin granular cells are visualized in real time. Release and tissue activity of renin can be studied on the individual granule level. Imaging of the living inner medulla shows how interstitial cells interconnect cells of the vasa recta, loop of Henle, and collecting duct. In summary, multiphoton microscopy is an exciting new optical sectioning technique that has great potential for numerous future developments and is ideal for applications that require deep optical sectioning of living tissue samples. PMID- 15883167 TI - Intravital multiphoton microscopy of dynamic renal processes. AB - Recent advances in microscopy and optics, computer sciences, and the available fluorophores used to label molecules of interest have empowered investigators to utilize intravital two-photon microscopy to study the dynamic events within the functioning kidney. This emerging technique enables investigators to follow functional and structural alterations with subcellular resolution within the same field of view over seconds to weeks. This approach invigorates the validity of data and facilitates analysis and interpretation as trends are more readily determined when one is more closely monitoring indicative physiological parameters. Therefore, in this review we emphasize how specific approaches will enable studies into glomerular permeability, proximal tubule endocytosis, and microvascular function within the kidney. We attempt to show how visual data can be quantified, thus allowing enhanced understanding of the process under study. Finally, emphasis is given to the possible future opportunities of this technology and its present limitations. PMID- 15883168 TI - Measuring DeltaPsim in isolated tubules. PMID- 15883169 TI - A cytomegaloviral protein reveals a dual role for STAT2 in IFN-{gamma} signaling and antiviral responses. AB - A mouse cytomegalovirus (MCMV) gene conferring interferon (IFN) resistance was identified. This gene, M27, encodes a 79-kD protein that selectively binds and down-regulates for signal transducer and activator of transcription (STAT)-2, but it has no effect on STAT1 activation and signaling. The absence of pM27 conferred MCMV susceptibility to type I IFNs (alpha/beta), but it had a much more dramatic effect on type II IFNs (gamma) in vitro and in vivo. A comparative analysis of M27(+) and M27(-) MCMV revealed that the antiviral efficiency of IFN-gamma was partially dependent on the synergistic action of type I IFNs that required STAT2. Moreover, STAT2 was directly activated by IFN-gamma. This effect required IFN receptor expression and was independent of type I IFNs. IFN-gamma induced increasing levels of tyrosine-phosphorylated STAT2 in M27(-) MCMV-infected cells that were essential for the antiviral potency of IFN-gamma. pM27 represents a new strategy for simultaneous evasions from types I and II IFNs, and it documents an unknown biological significance for STAT2 in antiviral IFN-gamma responses. PMID- 15883170 TI - Generation of hematopoietic repopulating cells from human embryonic stem cells independent of ectopic HOXB4 expression. AB - Despite the need for alternative sources of human hematopoietic stem cells (HSCs), the functional capacity of hematopoietic cells generated from human embryonic stem cells (hESCs) has yet to be evaluated and compared with adult sources. Here, we report that somatic and hESC-derived hematopoietic cells have similar phenotype and in vitro clonogenic progenitor activity. However, in contrast with somatic cells, hESC-derived hematopoietic cells failed to reconstitute intravenously transplanted recipient mice because of cellular aggregation causing fatal emboli formation. Direct femoral injection allowed recipient survival and resulted in multilineage hematopoietic repopulation, providing direct evidence of HSC function. However, hESC-derived HSCs had limited proliferative and migratory capacity compared with somatic HSCs that correlated with a distinct gene expression pattern of hESC-derived hematopoietic cells that included homeobox (HOX) A and B gene clusters. Ectopic expression of HOXB4 had no effect on repopulating capacity of hESC-derived cells. We suggest that limitations in the ability of hESC-derived HSCs to activate a molecular program similar to somatic HSCs may contribute to their atypical in vivo behavior. Our study demonstrates that HSCs can be derived from hESCs and provides an in vivo system and molecular foundation to evaluate strategies for the generation of clinically transplantable HSC from hESC lines. PMID- 15883171 TI - Decay-accelerating factor modulates induction of T cell immunity. AB - Decay-accelerating factor (Daf) dissociates C3/C5 convertases that assemble on host cells and thereby prevents complement activation on their surfaces. We demonstrate that during primary T cell activation, the absence of Daf on antigen presenting cells (APCs) and on T cells enhances T cell proliferation and augments the induced frequency of effector cells. The effect is factor D- and, at least in part, C5-dependent, indicating that local alternative pathway activation is essential. We show that cognate T cell-APC interactions are accompanied by rapid production of alternative pathway components and down-regulation of Daf expression. The findings argue that local alternative pathway activation and surface Daf protein function respectively as a costimulator and a negative modulator of T cell immunity and explain previously reported observations linking complement to T cell function. The results could have broad therapeutic implications for disorders in which T cell immunity is important. PMID- 15883172 TI - Recruitment of latent pools of high-avidity CD8(+) T cells to the antitumor immune response. AB - A major barrier to successful antitumor vaccination is tolerance of high-avidity T cells specific to tumor antigens. In keeping with this notion, HER-2/neu (neu) targeted vaccines, which raise strong CD8(+) T cell responses to a dominant peptide (RNEU(420-429)) in WT FVB/N mice and protect them from a neu-expressing tumor challenge, fail to do so in MMTV-neu (neu-N) transgenic mice. However, treatment of neu-N mice with vaccine and cyclophosphamide-containing chemotherapy resulted in tumor protection in a proportion of mice. This effect was specifically abrogated by the transfer of neu-N-derived CD4(+)CD25(+) T cells. RNEU(420-429)-specific CD8(+) T cells were identified only in neu-N mice given vaccine and cyclophosphamide chemotherapy which rejected tumor challenge. Tetramer-binding studies demonstrated that cyclophosphamide pretreatment allowed the activation of high-avidity RNEU(420-429)-specific CD8(+) T cells comparable to those generated from vaccinated FVB/N mice. Cyclophosphamide seemed to inhibit regulatory T (T reg) cells by selectively depleting the cycling population of CD4(+)CD25(+) T cells in neu-N mice. These findings demonstrate that neu-N mice possess latent pools of high-avidity neu-specific CD8(+) T cells that can be recruited to produce an effective antitumor response if T reg cells are blocked or removed by using approaches such as administration of cyclophosphamide before vaccination. PMID- 15883173 TI - The macrophage F4/80 receptor is required for the induction of antigen-specific efferent regulatory T cells in peripheral tolerance. AB - We show that the mouse macrophage-restricted F4/80 protein is not required for the development and distribution of tissue macrophages but is involved in the generation of antigen-specific efferent regulatory T (T reg) cells that suppress antigen-specific immunity. In the in vivo anterior chamber (a.c.)-associated immune deviation (ACAID) model of peripheral tolerance, a.c. inoculation of antigen into F4/80(-/-) mice was unable to induce efferent T reg cells and suppress delayed-type hypersensitivity (DTH) responses. Moreover, the use of anti F4/80 mAb and F4/80(-/-) APCs in an in vitro ACAID model showed that all APC cells in the culture must be able to express F4/80 protein if efferent T reg cells were to be generated. In a low-dose oral tolerance model, WT but not F4/80( /-) mice generated an efferent CD8(+) T reg cell population that suppressed an antigen-specific DTH response. Peripheral tolerance was restored in F4/80(-/-) mice by adoptive transfer of F4/80(+) APCs in both peripheral tolerance models, indicating a central role for the F4/80 molecule in the generation of efferent CD8(+) T reg cells. PMID- 15883174 TI - Citywide emergence of Pseudomonas aeruginosa strains with reduced susceptibility to polymyxin B. AB - OBJECTIVES: To determine the prevalence of Pseudomonas aeruginosa isolates with reduced susceptibility to polymyxin B, and to assess the in vitro activity of antibiotic combinations. METHODS: All unique patient isolates of P. aeruginosa were collected from 11 Brooklyn, NY hospitals during a three month period in 2003. Isolates with reduced susceptibility to polymyxin B (MIC > 2 mg/L) underwent ribotyping. The activity of polymyxin B combined with rifampicin, azithromycin and/or imipenem was tested by the chequerboard and time-kill methods against a subset of isolates. RESULTS: Of 527 isolates, only 61% were susceptible to imipenem. Twenty-five isolates (5%), from 8/11 hospitals, had reduced susceptibility to polymyxin B (MICs 4-8 mg/L), compared with 0/691 isolates collected in 2001. Ten of 25 were resistant to multiple other antibiotic classes. Ribotyping of the isolates revealed 19 unique types. Chequerboard testing of the 10 multiresistant isolates demonstrated synergy for the combinations of polymyxin B with azithromycin, imipenem and rifampicin in 6, 2, and 1 isolates, respectively. Time-kill studies revealed bactericidal activity for the following antibiotics when combined with polymyxin B: imipenem plus rifampicin against all 10 isolates, rifampicin in 9/10 isolates, imipenem in 8/10 isolates and azithromycin in 4/10 isolates. MICs of bacteria surviving incubation in polymyxin B alone rose for 4/9 isolates (MIC range 12-48 mg/L). CONCLUSIONS: P. aeruginosa with reduced susceptibility to polymyxin B have emerged in multiple strains in Brooklyn, NY. Combinations of polymyxin B with rifampicin and/or imipenem are bactericidal. The clinical utility of these combinations remains to be determined. PMID- 15883175 TI - Phenotypic variability of Pseudomonas aeruginosa in sputa from patients with acute infective exacerbation of cystic fibrosis and its impact on the validity of antimicrobial susceptibility testing. AB - OBJECTIVES: To investigate the variability in antimicrobial susceptibility of Pseudomonas aeruginosa from sputa of patients with cystic fibrosis, to compare testing individual colonies of the same morphotype either separately or combined and to study the reproducibility of testing antimicrobial susceptibility within and between laboratories. METHODS: One hundred and one sputa were cultured. Four colonies of each P. aeruginosa morphotype were suspended. Susceptibility to 12 agents by disc diffusion was tested individually or by pooling the four suspensions. A sputum sample containing four morphotypes of one genotype of P. aeruginosa was used to study reproducibility. Susceptibility was tested in duplicate by eight biomedical scientists in one laboratory and by routine procedures in seven different laboratories. RESULTS: There was a mean of four morphotypes of P. aeruginosa per sputum and three antibiograms per morphotype. In some cases, all four colonies of a single morphotype had different antibiograms. The susceptibility profiles of single isolates of P. aeruginosa correlated poorly with pooled cultures, with the pooled tests missing resistance. Results from one sample tested in duplicate by eight biomedical scientists in one laboratory and in seven other laboratories did not correlate well. The wide range of zone sizes in disc diffusion tests illustrated the variation in susceptibility of 48 colonies from one sputum sample. CONCLUSIONS: The role of conventional antimicrobial susceptibility testing is questionable once P. aeruginosa chronically infects the cystic fibrosis lung. A range of susceptibility patterns is seen, even within a morphotype. Routine test results are not reproducible and underestimate resistance. PMID- 15883176 TI - Gentamicin-loaded microspheres for treatment of experimental Brucella abortus infection in mice. AB - OBJECTIVES: To evaluate the efficacy of gentamicin-loaded poly (lactide-co glycolide) 50:50H (PLGA 50:50H) microspheres for the treatment of mice experimentally infected with Brucella abortus 2308. METHODS: The microspheres were dispersed in either 2% (w/v) poloxamer 188 saline solution, or deionized water with the help of a cell homogenizer to break up particle aggregates, and were administered intravenously or intraperitoneally to B. abortus-infected mice 7 days post-infection. RESULTS: Neither a single intravenous or intraperitoneal dose of 67 microg of gentamicin per mouse, nor three intraperitoneal doses of 100 microg of gentamicin per mouse, reduced the Brucella infection in the spleen compared with untreated mice 1 and 3 weeks post-treatment. Histological examination revealed granulation and tissue reaction in the periphery of spleen and liver of animals given three doses of the gentamicin-loaded microspheres. CONCLUSIONS: The lack of therapeutic activity of the gentamicin-loaded microspheres might be related to inappropriate microsphere size and aggregation, resulting also in a poor distribution of the microspheres in the spleen. The results might provide an example of practical problems related to particle size and aggregation for in vivo therapy with PLGA microspheres. PMID- 15883177 TI - Low-doses of indinavir boosted with ritonavir in HIV-infected Thai patients: pharmacokinetics, efficacy and tolerability. AB - OBJECTIVES: To assess the steady-state pharmacokinetics of two reduced doses of indinavir boosted with ritonavir (indinavir/ritonavir) in HIV-infected Thai patients. PATIENTS AND METHODS: Thirteen immunocompromised antiretroviral-naive patients (6 males, 7 females) initiated 600/100 mg indinavir/ritonavir, zidovudine and lamivudine, every 12 h. After 1 month, blood samples were taken at pre-dose, and 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 8 and 12 h after drug intake. Indinavir dosing was then reduced to 400 mg (twice daily) and 1 week later an identical series of samples were drawn. Patients then resumed 600 mg of indinavir. HIV-1 RNA viral load was determined at 8, 24 and 48 weeks. Indinavir plasma levels were determined by HPLC and pharmacokinetic parameters by non compartmental analysis. RESULTS: Median (range) weight was 58 kg (51-73) for men and 53 kg (46-59) for women. On 600 mg of indinavir, median indinavir AUC, C(max), and C(min) were 39.3 mg.h/L (20.6-50.5), 6.2 mg/L (3.7-9.0) and 0.41 mg/L (0.12-0.77), respectively, and on indinavir 400 mg, 18.3 mg.h/L (11.1-33.0), 3.8 mg/L (2.2-7.8) and 0.17 mg/L (0.10-0.39), respectively. No renal complications were observed. At 48 weeks, 6/13 (46%) patients had stopped 600 mg of indinavir due to intolerability (gastrointestinal and cutaneous), and 5/7 (71%) patients had a HIV-1 viral load <50 copies/mL. CONCLUSIONS: Reduced doses of indinavir/ritonavir maintained adequate indinavir plasma levels compared to current guidelines suggesting that these doses are efficacious in this setting. Considering the poor tolerability of 600 mg of indinavir, the 400 mg of indinavir may be preferred due to its lower exposure indices but long-term efficacy data are needed. PMID- 15883178 TI - Antibiotic susceptibilities of Legionella pneumophila strain Paris in THP-1 cells as determined by real-time PCR assay. AB - OBJECTIVES: Legionella species are facultative intracellular bacteria. Evaluation of the activity of antibiotics against intracellular L. pneumophila is more predictive of their in vivo efficacy than MICs as determined in axenic medium. However, current methodologies are based on cfu count determination, and are tedious because of the slow growth of Legionella spp. We investigated antibiotic susceptibilities of L. pneumophila strain Paris in THP-1-derived macrophages, using a real-time PCR assay for evaluation of bacterial growth. METHODS: Intracellular activities of seven antibiotic compounds against two human isolates of L. pneumophila strain Paris were determined in THP-1-derived macrophages in vitro. Bacterial growth was evaluated using either cfu methodology or a real-time PCR protocol targeting the mip gene. RESULTS: Bacterial titres as determined using real-time PCR were well correlated with cfu counts. Antibiotic susceptibilities for the two L. pneumophila isolates tested were comparable when using either of the two techniques. MICs were also similar to those previously reported for other L. pneumophila serogroup 1 strains. In particular, rifampicin and the fluoroquinolones were the most active compounds, both in extracellular medium and in THP-1 cells. Real-time PCR, however, was much less laborious than the traditional cfu method. CONCLUSIONS: Real-time PCR is better adapted than cfu based methods to evaluating the antibiotic susceptibilities of large series of Legionella strains to newer antibiotic compounds. PMID- 15883179 TI - Effect of media composition and in vitro activity of posaconazole, caspofungin and voriconazole against zygomycetes. AB - OBJECTIVES: The effect of different media and composition on the in vitro activity of posaconazole, caspofungin and voriconazole against 59 zygomycetes species was determined. METHODS: The media tested were RPMI 1640 medium with and without 2% glucose, antibiotic medium 3 (AM3) with and without 2% glucose, and high resolution (HR) medium. RESULTS: Posaconazole was significantly more active than caspofungin and voriconazole, both in RPMI 1640 medium with 2% glucose and in HR medium. Adding glucose improved the determination of end points, but had only minor influence on the MICs. MICs evaluated in AM3 were lower than in RPMI 1640 medium or HR medium. CONCLUSIONS: The in vivo effect of posaconazole in zygomycosis needs further evaluation. PMID- 15883180 TI - Brevundimonas diminuta infections and its resistance to fluoroquinolones. AB - OBJECTIVES: To report infections caused by Brevundimonas diminuta and antibiotic studies of this Gram-negative bacterium. PATIENTS AND METHODS: Seven patients with infection and eight bacterial strains were studied. Tests included antibiotic susceptibility and analysis of the DNA gyrase and topoisomerase genes and the effect of efflux pump inhibitor Phe-Arg-beta-naphthylamide (PANA). RESULTS: The patients all had underlying disease of cancer. The infections involved bloodstream (one case), intravascular catheter (four cases), urinary tract (one case) and pleural space (one case of empyema). Fever up to 39.2 degrees C characterized these infections, which resolved upon treatment by combination antibiotics. Microbiologically, all organisms were resistant to multiple fluoroquinolones and cefepime, but were susceptible to amikacin, imipenem and ticarcillin/clavulanate. These quinolone-resistant B. diminuta strains were probably selected out by the prophylactic use of a quinolone in six of these patients. Additionally, the B. diminuta type strain ATCC 11568(T) that was isolated before the quinolone era from water was also resistant to ciprofloxacin and intermediate to levofloxacin, suggesting intrinsic quinolone resistance. The DNA gyrase and topoisomerase of six analysed strains all contained GyrA Ala-83 and Met-87, GyrB Leu-466 or Thr-466, and ParC Gln-57, Val 66 and Ala-80 that were probably the cause of fluoroquinolone resistance. PANA had nearly negligible effect. CONCLUSIONS: B. diminuta is intrinsically resistant to fluoroquinolones and can be selected out to cause infections. PMID- 15883181 TI - The influence of phospholipid membranes on bovine calcitonin secondary structure and amyloid formation. AB - Calcitonin, a peptide hormone associated with medullary carcinoma of the thyroid, has the potential to form amyloid fibrils and may be a valuable model for investigating the role of peptide-membrane interactions in beta-sheet and amyloid formation. Via a new model peptide system, bovine calcitonin, we found that the exposure of peptide to phospholipid membranes altered its structure relative to the structures formed in aqueous solutions. Of particular relevance to the amyloidoses, incubation of calcitonin with cholesterol-rich and ganglioside containing membranes resulted in significant enrichment in the beta-sheet and amyloid content of the peptide. The formation of amyloid was also accelerated in these systems. A correlation between the phospholipid-induced structural alterations and calcitonin binding affinities to phospholipid membranes was evident. Bovine calcitonin has considerably higher binding affinity for the phospholipid systems that enhanced its beta-sheet and amyloid structure. Electrostatic forces were not the governing forces behind the observed behavior, as supported by the fact that the ionic strength did not affect the peptide structures or binding affinities. A Van't Hoff analysis of the temperature dependent peptide binding affinities indicated that binding led to an increase in enthalpy and possibly an increase in entropy of the peptide-membrane systems. Experiments with other amyloid-forming peptides such as beta-amyloid of Alzheimer's disease have also shown similar results and may indicate the need to manipulate peptide-membrane interactions in order to control amyloid formation and its associated disease. PMID- 15883182 TI - HU-alpha binds to the putative double-stranded DNA mimic HI1450 from Haemophilus influenzae. AB - Recently, the solution structure of the hypothetical protein HI1450 from Haemophilus influenzae was solved as part of a structure-based effort to understand function. The distribution of its many negatively charged residues and weak structure and sequence homology to uracil DNA glycosylase inhibitor (Ugi) suggested that HI1450 may act as a double-stranded DNA (dsDNA) mimic. We present supporting evidence here and show that HI1450 interacts with the dsDNA-binding protein HU-alpha. The interaction between HI1450 and HU-alpha from H. influenzae is characterized using calorimetry and NMR spectroscopy. HU-alpha binds to HI1450 with a K(d) of 3.0 +/- 0.2 microM, which is similar in affinity to its interaction with dsDNA. Chemical shift perturbation data indicate that the beta1 strand of HI1450 and neighboring regions are most directly involved in interactions with HU-alpha. These results show that HI1450 and its structural homolog, Ugi, use similar parts of their structures to recognize DNA-binding proteins. PMID- 15883183 TI - p25alpha is flexible but natively folded and binds tubulin with oligomeric stoichiometry. AB - p25alpha is a 219-residue protein which stimulates aberrant tubulin polymerization and is implicated in a variety of other functions. The protein has unusual secondary structure involving significant amounts of random coil, and binding to microtubules is accompanied by a large structural change, suggesting a high degree of plasticity. p25alpha has been proposed to be natively unfolded, so that folding is coupled to interaction with its physiological partners. Here we show that recombinant human p25alpha is folded under physiological conditions, since it has a well structured and solvent-sequestered aromatic environment and considerable chemical shift dispersion of amide and aliphatic protons. With increasing urea concentrations, p25alpha undergoes clear spectral changes suggesting significant loss of structure. p25alpha unfolds cooperatively in urea according to a simple two-state transition with a stability in water of approximately 5 kcal/mol. The protein behaves as a monomer and refolds with a transient on-pathway folding intermediate. However, high sensitivity to proteolytic attack and abnormal gel filtration migration behavior suggests a relatively extended structure, possibly organized in distinct domains. A deletion mutant of p25alpha lacking residues 3-43 also unfolds cooperatively and with similar stability, suggesting that the N-terminal region is largely unstructured. Both proteins undergo significant loss of structure when bound to monomeric tubulin. The stoichiometry of binding is estimated to be 3-4 molecules of tubulin per p25alpha and is not significantly affected by the deletion of residues 3-43. In conclusion, we dismiss the proposal that p25alpha is natively unfolded, although the protein is relatively flexible. This flexibility may be linked to its tubulin-binding properties. PMID- 15883184 TI - Mass spectrometric analysis of the human 40S ribosomal subunit: native and HCV IRES-bound complexes. AB - Hepatitis C virus uses an internal ribosome entry site (IRES) in the viral RNA to directly recruit human 40S ribosome subunits during cap-independent translation initiation. Although IRES-mediated translation initiation is not subject to many of the regulatory mechanisms that control cap-dependent translation initiation, it is unknown whether other noncanonical protein factors are involved in this process. Thus, a global protein composition analysis of native and IRES-bound 40S ribosomal complexes has been conducted to facilitate an understanding of the IRES ribosome recruitment mechanism. A combined top-down and bottom-up mass spectrometry approach was used to identify both the proteins and their posttranslational modifications (PTMs) in the native 40S subunit and the IRES recruited translation initiation complex. Thirty-one out of a possible 32 ribosomal proteins were identified by combining top-down and bottom-up mass spectrometry techniques. Proteins were found to contain PTMs, including loss of methionine, acetylation, methylation, and disulfide bond formation. In addition to the 40S ribosomal proteins, RACK1 was consistently identified in the 40S fraction, indicating that this protein is associated with the 40S subunit. Similar methodology was then applied to the hepatitis C virus IRES-bound 40S complex. Two 40S ribosomal proteins, RS25 and RS29, were found to contain different PTMs than those in the native 40S subunit. In addition, RACK1, eukaryotic initiation factor 3 proteins and nucleolin were identified in the IRES mediated translation initiation complex. PMID- 15883185 TI - Novel and economical purification of recombinant proteins: intein-mediated protein purification using in vivo polyhydroxybutyrate (PHB) matrix association. AB - This work combines two well-established technologies to generate a breakthrough in protein production and purification. The first is the production of polyhydroxybutyrate (PHB) granules in engineered strains of Escherichia coli. The second is a recently developed group of self-cleaving affinity tags based on protein splicing elements known as inteins. By combining these technologies with a PHB-specific binding protein, a self-contained protein expression and purification system has been developed. In this system, the PHB-binding protein effectively acts as an affinity tag for desired product proteins. The tagged product proteins are expressed in E. coli strains that also produce intracellular PHB granules, where they bind to the granules via the PHB-binding tag. The granules and attached proteins can then be easily recovered following cell lysis by simple mechanical means. Once purified, the product protein is self-cleaved from the granules and released into solution in a substantially purified form. This system has been successfully used at laboratory scale to purify several active test proteins at reasonable yield. By allowing the bacterial cells to effectively produce both the affinity resin and tagged target protein, the cost associated with the purification of recombinant proteins could be greatly reduced. It is expected that this combination of improved economics and simplicity will constitute a significant breakthrough in both large-scale production of purified proteins and enzymes and high-throughput proteomics studies of peptide libraries. PMID- 15883186 TI - Domain swapping in the low-similarity isomerase/hydratase superfamily: the crystal structure of rat mitochondrial Delta3, Delta2-enoyl-CoA isomerase. AB - Two monofunctional Delta(3), Delta(2)-enoyl-CoA isomerases, one in mitochondria (mECI) and the other in both mitochondria and peroxisomes (pECI), belong to the low-similarity isomerase/hydratase superfamily. Both enzymes catalyze the movement of a double bond from C3 to C2 of an unsaturated acyl-CoA substrate for re-entry into the beta-oxidation pathway. Mutagenesis has shown that Glu165 of rat mECI is involved in catalysis; however, the putative catalytic residue in yeast pECI, Glu158, is not conserved in mECI. To elucidate whether Glu165 of mECI is correctly positioned for catalysis, the crystal structure of rat mECI has been solved. Crystal packing suggests the enzyme is trimeric, in contrast to other members of the superfamily, which appear crystallographically to be dimers of trimers. The polypeptide fold of mECI, like pECI, belongs to a subset of this superfamily in which the C-terminal domain of a given monomer interacts with its own N-terminal domain. This differs from that of crotonase and 1,4-dihydroxy-2 naphtoyl-CoA synthase, whose C-terminal domains are involved in domain swapping with an adjacent monomer. The structure confirms Glu165 as the putative catalytic acid/base, positioned to abstract the pro-R proton from C2 and reprotonate at C4 of the acyl chain. The large tunnel-shaped active site cavity observed in the mECI structure explains the relative substrate promiscuity in acyl-chain length and stereochemistry. Comparison with the crystal structure of pECI suggests the catalytic residues from both enzymes are spatially conserved but not in their primary structures, providing a powerful reminder of how catalytic residues cannot be determined solely by sequence alignments. PMID- 15883187 TI - Calcium- and magnesium-dependent interactions between calcium- and integrin binding protein and the integrin alphaIIb cytoplasmic domain. AB - Calcium- and integrin-binding protein (CIB) is a small EF-hand calcium-binding protein that is involved in hemostasis through its interaction with the alphaIIb cytoplasmic domain of integrinalphaIIbbeta(3). We have previously demonstrated that CIB lacks structural stability in the absence of divalent metal ions but that it acquires a well-folded conformation upon addition of Ca(2+) or Mg(2+). Here, we have used fluorescence spectroscopy, NMR spectroscopy, and isothermal titration calorimetry to demonstrate that both Ca(2+)-bound CIB (Ca(2+)-CIB) and the Mg(2+)-bound protein (Mg(2+)-CIB) bind with high affinity and through a similar mechanism to alphaIIb cytoplasmic domain peptides, but that metal-free CIB (apo-CIB) binds in a different manner. The interactions are thermodynamically distinct for Ca(2+)-CIB and Mg(2+)-CIB, but involve hydrophobic interactions in each case. Since the Mg(2+) concentration inside the cell is sufficient to saturate CIB at all times, our results imply that CIB would be capable of binding to the alphaIIb cytoplasmic domain independent of an intracellular Ca(2+) stimulus in vivo. This raises the question of whether CIB can act as a Ca(2+) sensor in alphaIIbbeta(3) signaling or if other regulatory mechanisms such as fibrinogen-induced conformational changes in alphaIIbbeta(3), post-translational modifications, or the binding of other accessory proteins mediate the interactions between CIB and alphaIIbbeta(3). Differences in NMR spectra do suggest, however, that Ca(2+)-binding to the Mg(2+)- CIB-alphaIIb complex induces subtle structural changes that could further modulate the activity of alphaIIbbeta(3). PMID- 15883188 TI - The solution structure of the oxidative stress-related protein YggX from Escherichia coli. AB - YggX is a highly conserved protein found only in eubacteria and is proposed to be involved in the bacterial response to oxidative stress. Here we report the solution structure of YggX from Escherichia coli determined by nuclear magnetic resonance spectroscopy. The structure of YggX displays a fold consisting of two N terminal antiparallel beta-sheets and three alpha-helices, which shares significant structural similarity to the crystal structure of a hypothetical protein PA5148 from Pseudomonas aeruginosa. Previous studies propose YggX as an iron binding protein that is involved in cellular iron trafficking. Our data indicate that the protein alone does not bind iron in vitro, suggesting other cofactors or different conditions may be necessary for metal binding. PMID- 15883189 TI - Asymmetric effect of domain interactions on the kinetics of folding in yeast phosphoglycerate kinase. AB - The aim of this work is to shed more light on the effect of domain-domain interactions on the kinetics and the pathway of protein folding. A model protein system consisting of several single-tryptophan variants of the two-domain yeast phosphoglycerate kinase (PGK) and its individual domains was studied. Refolding was initiated from the guanidine-unfolded state by stopped-flow or manual mixing and monitored by tryptophan fluorescence from 1 msec to 1000 sec. Denaturant titrations of both individual domains showed apparent two-state unfolding transitions. Refolding kinetics of the individual domains from different denaturant concentrations, however, revealed the presence of intermediate structures during titration for both domains. Refolding of the same domains within the complete protein showed that domain-domain interactions direct the folding of both domains, but in an asymmetric way. Folding of the N domain was already altered within 1 msec, while detectable changes in the folding of the C domain occurred only 60-100 msec after initiating refolding. All mutants showed a hyperfluorescent kinetic intermediate. Both the disappearance of this intermediate and the completion of the folding were significantly faster in the individual N domain than in the complete protein. On the contrary, folding of the individual C domain was slower than in the complete protein. The presence of the C domain directs the refolding of the N domain along a completely different pathway than that of the individual N domain, while folding of the individual C domain follows the same path as within the complete protein. PMID- 15883190 TI - Capsid structure and dynamics of a human rhinovirus probed by hydrogen exchange mass spectrometry. AB - Viral capsids are dynamic protein assemblies surrounding viral genomes. Despite the high-resolution structures determined by X-ray crystallography and cryo electron microscopy, their in-solution structure and dynamics can be probed by hydrogen exchange. We report here using hydrogen exchange combined with protein enzymatic fragmentation and mass spectrometry to determine the capsid structure and dynamics of a human rhinovirus, HRV14. Capsid proteins (VP1-4) were labeled with deuterium by incubating intact virus in D(2)O buffer at neutral pH. The labeled proteins were digested by immobilized pepsin to give peptides analyzed by capillary reverse-phase HPLC coupled with nano-electrospray mass spectrometry. Deuterium levels incorporated at amide linkages in peptic fragments were measured for different exchange times from 12 sec to 30 h to assess the amide hydrogen exchange rates along each of the four protein backbones. Exchange results generally agree with the crystal structure of VP1-4,with extended, flexible terminal and surface-loop regions in fast exchange and folded helical and sheet structures in slow exchange. In addition, three alpha-helices, one from each of VP1-3, exhibited very slow exchange, indicating high stability of the protomeric interface. The beta-strands at VP3 N terminus also had very slow exchange, suggesting stable pentamer contacts. It was noted, however, that the interface around the fivefold axis had fast and intermediate exchange, indicating relatively more flexibility. Even faster exchange rates were found in the N terminus of VP1 and most segments of VP4, suggesting high flexibilities, which may correspond to their potential roles in virus uncoating. PMID- 15883191 TI - Strain relief at the active site of phosphoserine aminotransferase induced by radiation damage. AB - The X-ray susceptibility of the lysine-pyridoxal-5'-phosphate Schiff base in Bacillus alcalophilus phosphoserine aminotransferase has been investigated using crystallographic data collected at 100 K to 1.3 A resolution, complemented by on line spectroscopic studies. X-rays induce deprotonation of the internal aldimine, changes in the Schiff base conformation, displacement of the cofactor molecule, and disruption of the Schiff base linkage between pyridoxal-5'-phosphate and the Lys residue. Analysis of the "undamaged" structure reveals a significant chemical strain on the internal aldimine bond that leads to a pronounced geometrical distortion of the cofactor. However, upon crystal exposure to the X-rays, the strain and distortion are relaxed and eventually diminished when the total absorbed dose has exceeded 4.7 x 10(6) Ggamma. Our data provide new insights into the enzymatic activation of pyridoxal-5'-phosphate and suggest that special care should be taken while using macromolecular crystallography to study details in strained active sites. PMID- 15883192 TI - Sensory nerve inactivation by resiniferatoxin improves insulin sensitivity in male obese Zucker rats. AB - Recent studies have suggested that sensory nerves may influence insulin secretion and action. The present study investigated the effects of resiniferatoxin (RTX) inactivation of sensory nerves (desensitization) on oral glucose tolerance, insulin secretion and whole body insulin sensitivity in the glucose intolerant, hyperinsulinemic, and insulin-resistant obese Zucker rat. After RTX treatment (0.05 mg/kg RTX sc given at ages 8, 10, and 12 wk), fasting plasma insulin was reduced (P < 0.0005), and oral glucose tolerance was improved (P < 0.005). Pancreas perfusion showed that baseline insulin secretion (7 mM glucose) was lower in RTX-treated rats (P = 0.01). Insulin secretory responsiveness to 20 mM glucose was enhanced in the perfused pancreas of RTX-treated rats (P < 0.005) but unaffected in stimulated, isolated pancreatic islets. At the peak of spontaneous insulin resistance in the obese Zucker rat, insulin sensitivity was substantially improved after RTX treatment, as evidenced by higher glucose infusion rates (GIR) required to maintain euglycemia during a hyperinsulinemic euglycemic (5 mU.kg( 1).min(-1)) clamp (GIR(60-120min): 5.97 +/- 0.62 vs. 11.65 +/- 0.83 mg.kg( 1).min(-1) in RTX-treated rats, P = 0.003). In conclusion, RTX treatment and, hence, sensory nerve desensitization of adult male obese Zucker rats improved oral glucose tolerance by enhancing insulin secretion, and, in particular, by improving insulin sensitivity. PMID- 15883193 TI - MCAK associates with the tips of polymerizing microtubules. AB - MCAK is a member of the kinesin-13 family of microtubule (MT)-depolymerizing kinesins. We show that the potent MT depolymerizer MCAK tracks (treadmills) with the tips of polymerizing MTs in living cells. Tip tracking of MCAK is inhibited by phosphorylation and is dependent on the extreme COOH-terminal tail of MCAK. Tip tracking is not essential for MCAK's MT-depolymerizing activity. We propose that tip tracking is a mechanism by which MCAK is preferentially localized to regions of the cell that modulate the plus ends of MTs. PMID- 15883194 TI - Molecular constituents of neuronal AMPA receptors. AB - Dynamic regulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) underlies aspects of synaptic plasticity. Although numerous AMPAR-interacting proteins have been identified, their quantitative and relative contributions to native AMPAR complexes remain unclear. Here, we quantitated protein interactions with neuronal AMPARs by immunoprecipitation from brain extracts. We found that stargazin-like transmembrane AMPAR regulatory proteins (TARPs) copurified with neuronal AMPARs, but we found negligible binding to GRIP, PICK1, NSF, or SAP-97. To facilitate purification of neuronal AMPAR complexes, we generated a transgenic mouse expressing an epitope-tagged GluR2 subunit of AMPARs. Taking advantage of this powerful new tool, we isolated two populations of GluR2 containing AMPARs: an immature complex with the endoplasmic reticulum chaperone immunoglobulin-binding protein and a mature complex containing GluR1, TARPs, and PSD-95. These studies establish TARPs as the auxiliary components of neuronal AMPARs. PMID- 15883195 TI - Promotion of importin alpha-mediated nuclear import by the phosphorylation dependent binding of cargo protein to 14-3-3. AB - 14-3-3 proteins are phosphoserine/threonine-binding proteins that play important roles in many regulatory processes, including intracellular protein targeting. 14 3-3 proteins can anchor target proteins in the cytoplasm and in the nucleus or can mediate their nuclear export. So far, no role for 14-3-3 in mediating nuclear import has been described. There is also mounting evidence that nuclear import is regulated by the phosphorylation of cargo proteins, but the underlying mechanism remains elusive. Myopodin is a dual-compartment, actin-bundling protein that functions as a tumor suppressor in human bladder cancer. In muscle cells, myopodin redistributes between the nucleus and the cytoplasm in a differentiation dependent and stress-induced fashion. We show that importin alpha binding and the subsequent nuclear import of myopodin are regulated by the serine/threonine phosphorylation-dependent binding of myopodin to 14-3-3. These results establish a novel paradigm for the promotion of nuclear import by 14-3-3 binding. They provide a molecular explanation for the phosphorylation-dependent nuclear import of nuclear localization signal-containing cargo proteins. PMID- 15883196 TI - Kinesin-1 mediates translocation of the meiotic spindle to the oocyte cortex through KCA-1, a novel cargo adapter. AB - In animals, female meiotic spindles are attached to the egg cortex in a perpendicular orientation at anaphase to allow the selective disposal of three haploid chromosome sets into polar bodies. We have identified a complex of interacting Caenorhabditis elegans proteins that are involved in the earliest step in asymmetric positioning of anastral meiotic spindles, translocation to the cortex. This complex is composed of the kinesin-1 heavy chain orthologue, UNC 116, the kinesin light chain orthologues, KLC-1 and -2, and a novel cargo adaptor, KCA-1. Depletion of any of these subunits by RNA interference resulted in meiosis I metaphase spindles that remained stationary at a position several micrometers from the cell cortex during the time when wild-type spindles translocated to the cortex. After this prolonged stationary period, unc-116(RNAi) spindles moved to the cortex through a partially redundant mechanism that is dependent on the anaphase-promoting complex. This study thus reveals two sequential mechanisms for translocating anastral spindles to the oocyte cortex. PMID- 15883197 TI - Spatial distribution and functional significance of activated vinculin in living cells. AB - Conformational change is believed to be important to vinculin's function at sites of cell adhesion. However, nothing is known about vinculin's conformation in living cells. Using a Forster resonance energy transfer probe that reports on changes in vinculin's conformation, we find that vinculin is in the actin-binding conformation in a peripheral band of adhesive puncta in spreading cells. However, in fully spread cells with established polarity, vinculin's conformation is variable at focal adhesions. Time-lapse imaging reveals a gradient of conformational change that precedes loss of vinculin from focal adhesions in retracting regions. At stable or protruding regions, recruitment of vinculin is not necessarily coupled to the actin-binding conformation. However, a different measure of vinculin conformation, the recruitment of vinexin beta by activated vinculin, shows that autoinhibition of endogenous vinculin is relaxed at focal adhesions. Beyond providing direct evidence that vinculin is activated at focal adhesions, this study shows that the specific functional conformation correlates with regional cellular dynamics. PMID- 15883198 TI - Regulation of phototransduction responsiveness and retinal degeneration by a phospholipase D-generated signaling lipid. AB - Drosophila melanogaster phototransduction proceeds via a phospholipase C (PLC) triggered cascade of phosphatidylinositol (PI) lipid modifications, many steps of which remain undefined. We describe the involvement of the lipid phosphatidic acid and the enzyme that generates it, phospholipase D (Pld), in this process. Pld(null) flies exhibit decreased light sensitivity as well as a heightened susceptibility to retinal degeneration. Pld overexpression rescues flies lacking PLC from light-induced, metarhodopsin-mediated degeneration and restores visual signaling in flies lacking the PI transfer protein, which is a key player in the replenishment of the PI 4,5-bisphosphate (PIP2) substrate used by PLC to transduce light stimuli into neurological signals. Altogether, these findings suggest that Pld facilitates phototransduction by maintaining adequate levels of PIP2 and by protecting the visual system from metarhodopsin-induced, low light degeneration. PMID- 15883199 TI - Galectin-4 and sulfatides in apical membrane trafficking in enterocyte-like cells. AB - We have previously reported that 1-benzyl-2-acetamido-2-deoxy-alpha-D galactopyranoside (GalNAc alpha-O-bn), an inhibitor of glycosylation, perturbed apical biosynthetic trafficking in polarized HT-29 cells suggesting an involvement of a lectin-based mechanism. Here, we have identified galectin-4 as one of the major components of detergent-resistant membranes (DRMs) isolated from HT-29 5M12 cells. Galectin-4 was also found in post-Golgi carrier vesicles. The functional role of galectin-4 in polarized trafficking in HT-29 5M12 cells was studied by using a retrovirus-mediated RNA interference. In galectin-4-depleted HT-29 5M12 cells apical membrane markers accumulated intracellularly. In contrast, basolateral membrane markers were not affected. Moreover, galectin-4 depletion altered the DRM association characteristics of apical proteins. Sulfatides with long chain-hydroxylated fatty acids, which were also enriched in DRMs, were identified as high-affinity ligands for galectin-4. Together, our data propose that interaction between galectin-4 and sulfatides plays a functional role in the clustering of lipid rafts for apical delivery. PMID- 15883200 TI - Agrin mediates a rapid switch from electrical coupling to chemical neurotransmission during synaptogenesis. AB - In contrast to its well-established actions as an organizer of synaptic differentiation at the neuromuscular junction, the proteoglycan agrin is still in search of a function in the nervous system. Here, we report an entirely unanticipated role for agrin in the dual modulation of electrical and chemical intercellular communication that occurs during the critical period of synapse formation. When applied at the developing splanchnic nerve-chromaffin cell cholinergic synapse in rat adrenal acute slices, agrin rapidly modified cell-to cell communication mechanisms. Specifically, it led to decreased gap junction mediated electrical coupling that preceded an increase in nicotinic synaptic transmission. This developmental switch from predominantly electrical to chemical communication was fully operational within one hour and depended on the activation of Src family-related tyrosine kinases. Hence, agrin may play a pivotal role in synaptogenesis in promoting a rapid switch between electrical coupling and synaptic neurotransmission. PMID- 15883202 TI - Molecular pharmacology of the interaction of anthracyclines with iron. AB - Although anthracyclines such as doxorubicin are widely used antitumor agents, a major limitation for their use is the development of cardiomyopathy at high cumulative doses. This severe adverse side effect may be due to interactions with cellular iron metabolism, because iron loading promotes anthracycline-induced cell damage. On the other hand, anthracycline-induced cardiotoxicity is significantly alleviated by iron chelators (e.g., desferrioxamine and dexrazoxane). The molecular mechanisms by which anthracyclines interfere with cellular iron trafficking are complex and still unclear. Doxorubicin can directly bind iron and can perturb iron metabolism by interacting with multiple molecular targets, including the iron regulatory proteins (IRP) 1 and 2. The RNA-binding activity of these molecules regulates synthesis of the transferrin receptor 1 and ferritin, which are crucial proteins involved in iron uptake and storage, respectively. At present, it is not clear whether doxorubicin affects IRP1-RNA binding activity by intracellular formation of doxorubicinol and/or by generation of the doxorubicin-iron(III) complex. Furthermore, doxorubicin prevents the mobilization of iron from ferritin by a mechanism that may involve lysosomal degradation of this protein. Prevention of iron mobilization from ferritin would probably disturb vital cellular functions as a result of inhibition of essential iron-dependent proteins, such as ribonucleotide reductase. This review discusses the molecular interactions of anthracyclines with iron metabolism and the development of cardioprotective strategies such as iron chelators. PMID- 15883201 TI - Tight junctions in Schwann cells of peripheral myelinated axons: a lesson from claudin-19-deficient mice. AB - Tight junction (TJ)-like structures have been reported in Schwann cells, but their molecular composition and physiological function remain elusive. We found that claudin-19, a novel member of the claudin family (TJ adhesion molecules in epithelia), constituted these structures. Claudin-19-deficient mice were generated, and they exhibited behavioral abnormalities that could be attributed to peripheral nervous system deficits. Electrophysiological analyses showed that the claudin-19 deficiency affected the nerve conduction of peripheral myelinated fibers. Interestingly, the overall morphology of Schwann cells lacking claudin-19 expression appeared to be normal not only in the internodal region but also at the node of Ranvier, except that TJs completely disappeared, at least from the outer/inner mesaxons. These findings have indicated that, similar to epithelial cells, Schwann cells also bear claudin-based TJs, and they have also suggested that these TJs are not involved in the polarized morphogenesis but are involved in the electrophysiological "sealing" function of Schwann cells. PMID- 15883203 TI - Cyclooxygenase-2 inhibitors decrease vascular endothelial growth factor expression in colon cancer cells by enhanced degradation of Sp1 and Sp4 proteins. AB - Cyclooxygenase 2 (COX-2) inhibitors, such as celecoxib (Cel), nimesulfide (NM), and NS-398 [NS; N-[2-(cyclohexyloxy)-4-nitrophenyl]methanesulfonamide] and other nonsteroidal anti-inflammatory drugs inhibit colon cancer growth and angiogenesis; however, the mechanism of this response is not well defined. Treatment of SW-480 colon cancer cells with Cel, NS, or NM decreased vascular endothelial growth factor (VEGF) mRNA and immunoreactive protein expression. This was also accompanied by decreased transactivation in cells transfected with constructs containing VEGF gene promoter inserts. Deletion analysis of the VEGF promoter indicated that decreased VEGF expression by COX-2 inhibitors was associated with the proximal -131 to -47 GC-rich region of the VEGF promoter that binds Sp proteins. Treatment of SW-480 cells with Cel, NM, and NS also decreased Sp1 and Sp4 protein expression but not that of Sp2 or Sp3. Similar results were observed in RKO, HT-29, and DLD colon cancer cells demonstrating comparable responses in COX-2-expressing and -nonexpressing colon cancer cell lines. COX-2 inhibitors do not affect Sp1 or Sp4 mRNA levels in SW-480 cells; however, decreased expression of both proteins was accompanied by increased protein ubiquitination and inhibited by the proteasome inhibitor gliotoxin. These results suggest that the antiangiogenic activity of COX-2 inhibitors in colon cancer cells is linked to activation of proteasome-dependent degradation of Sp1 and Sp4 proteins. PMID- 15883204 TI - A single residue in the S6 transmembrane domain governs the differential flecainide sensitivity of voltage-gated potassium channels. AB - Flecainide has been used to differentiate Kv4.2-based transient-outward K(+) currents (flecainide-sensitive) from Kv1.4-based (flecainide-insensitive). We found that flecainide also inhibits ultrarapid delayed rectifier (I(Kur)) currents in Xenopus laevis oocytes carried by Kv3.1 subunits (IC(50), 28.3 +/- 1.3 microM) more strongly than Kv1.5 currents corresponding to human I(Kur) (IC(50), 237.1 +/- 6.2 microM). The present study examined molecular motifs underlying differential flecainide sensitivity. An initial chimeric approach pointed to a role for S6 and/or carboxyl-terminal sites in Kv3.1/Kv1.5 sensitivity differences. We then looked for homologous amino acid residues of the two sensitive subunits (Kv4.2 and Kv3.1) different from homologous residues for insensitive subunits (Kv1.4 and Kv1.5). Three candidate sites were identified: two in the S5-S6 linker and one in the S6 segment. Mutation of the proximal S5-S6 linker site failed to alter flecainide sensitivity. Mutation at the more distal site in Kv1.5 (V481L) modestly increased sensitivity, but the reciprocal Kv3.1 mutation (L401V) had no effect. S6 mutants caused marked changes: flecainide sensitivity decreased approximately 8-fold for Kv3.1 L422I (IC(50), 213 +/- 9 microM) and increased approximately 7-fold for Kv1.5 I502L (IC(50), 35.6 +/- 1.9 microM). Corresponding mutations reversed flecainide sensitivity of Kv1.4 and Kv4.2; L392I decreased Kv4.2 sensitivity by approximately 17-fold (IC(50) of 37.4 +/- 6.9 to 628 +/- 36 microM); I547L increased Kv1.4 sensitivity by approximately 15-fold (IC(50) of 706 +/- 37 to 40.9 +/- 7.3 microM). Our observations indicate that the flecainide sensitivity differences among these four voltage-gated K(+) channels are determined by whether an isoleucine or a leucine is present at a specific amino acid location. PMID- 15883205 TI - Expansion of circulating Toll-like receptor 4-positive monocytes in patients with acute coronary syndrome. AB - BACKGROUND: Atherosclerosis is an inflammatory disease in which monocytes and macrophages have been suggested to play an essential role. The underlying signaling mechanisms are unknown thus far. We hypothesized that the human isoform of Toll-like receptor (hTLR)-4 is involved in monocyte activation of patients with accelerated forms of atherosclerosis. METHODS AND RESULTS: Expression of hTLR4 on circulating monocytes from 30 controls, 20 patients with stable angina (SA), 40 patients with unstable angina (UA), and 28 patients with acute myocardial infarction (AMI) was compared with the use of flow-cytometry and reverse transcription-polymerase chain reaction. Regulation of interleukin (IL) 12 and B7-1 as downstream events of TLR4 activation was analyzed after lipopolysaccharide stimulation of monocytes. TLR4-transfected Chinese hamster ovary (CHO) cells were used to identify potential hTLR4 ligands in the serum of patients with UA or AMI. Circulating hTLR4+/CD14+ monocytes were approximately 2.5-fold increased above controls and patients with SA in the UA and AMI groups (P<0.0001). This was paralleled by enhanced transcript levels of TLR4 and Myd88 in patients with UA and AMI (P<0.0001) and increased expression of IL-12 (UA 35.5+/-7.8, AMI 31.8+/-7.7 versus SA 2.2+/-0.5, controls 2.1+/-0.3 pg/mL; P<0.0002) and B7-1 (UA 27.3+/-14.4, AMI 22.6+/-11.1 versus SA 3.4+/-2.5, controls 2.4+/-2.3%; P<0.0001). Compared with serum from patients with UA and AMI, challenging TLR4-transfected CHO cells with serum from SA patients yielded only a weak response (P<0.0001). Coincubation with anti-heat shock protein 60 inhibited CHO cell activation. CONCLUSIONS: UA and AMI are associated with enhanced expression and signaling events downstream of hTLR4 in circulating monocytes. These observations suggest hTLR4 activation as a signaling mechanism in immune mediated progression of atherosclerosis. PMID- 15883206 TI - Insulin causes [Ca2+]i-dependent and [Ca2+]i-independent positive inotropic effects in failing human myocardium. AB - BACKGROUND: Insulin has been shown to exert positive inotropic effects in several in vitro and in vivo models, but signal transduction and substrate dependency remain unclear. We examined inotropic responses and signal transduction mechanisms of insulin in human myocardium. METHODS AND RESULTS: Experiments were performed in isolated trabeculae from end-stage failing hearts of 58 nondiabetic and 3 diabetic patients undergoing heart transplantation. The effect of insulin (0.3 and 3 IU/L) on isometric twitch force (37 degrees C, 1 Hz) was tested in the presence of glucose or pyruvate as energetic substrate. Furthermore, intracellular Ca2+ transients (aequorin method), sarcoplasmic reticulum (SR) Ca2+ content (rapid cooling contractures), and myofilament Ca2+ sensitivity (semiskinned fibers) were assessed. In addition, potential signaling pathways were tested by blocking glycolysis, PI-3-kinase, protein kinase C, diacylglycerol kinase, insulin-like growth factor-1 receptors, or transsarcolemmal Ca2+ entry via the Na+/Ca2+ exchanger. Insulin exerted concentration-dependent and partially substrate-dependent positive inotropic effects. The phosphatidylinositol-3-kinase inhibitor wortmannin and the Na2+/Ca2+ exchanger reverse-mode inhibitor KB-R7943 completely or partially prevented the functional effects of insulin. In contrast, insulin-like growth factor-1 receptor blockade, protein kinase C inhibition, and diacylglycerol kinase blockade were without effect. The inotropic response was associated with increases in intracellular Ca2+ transients, SR Ca2+ content, and increased myofilament Ca2+ sensitivity. CONCLUSIONS: Insulin exerts Ca2+ dependent and -independent positive inotropic effects through a phosphatidylinositol-3-kinase-dependent pathway in failing human myocardium. The increased [Ca2+]i originates at least in part from enhanced reverse-mode Na+/Ca2+ exchange and consequently increased SR-Ca2+ load. These nongenomic functional effects of insulin may be of clinical relevance, eg, during insulin-glucose potassium infusions. PMID- 15883207 TI - Endothelial progenitor cells are recruited into resolving venous thrombi. AB - BACKGROUND: The purpose of this study was to determine whether endothelial cells of bone marrow origin are involved in thrombus recanalization. METHODS AND RESULTS: Irradiated mice were reconstituted with bone marrow from transgenic donors expressing green fluorescent protein (GFP) linked to the Tie2 promoter. Thrombi were formed in 2 groups of 6 mice. GFP-expressing cells were located and quantified in sections of the thrombi taken after 7 and 14 days. The cell markers Mac-3, F4/80, CD68 (macrophage), and vascular endothelial growth factor receptor 2 (VEGFR2; endothelial cells) were used to determine colocalization with GFP expression in tissue sections and peritoneal macrophages. The markers CD34 and VEGFR2 were used to quantify changes in circulating endothelial cells by flow cytometry of blood from 3 cohorts of wild-type animals that had either a thrombus induced (n=18), a sham operation (n=18), or no operation (n=10). The number of GFP-expressing cells was found to increase by approximately 3-fold in thrombi formed in transplanted animals between 7 and 14 days after induction (P=0.0022). No GFP-expressing cells were found lining the new vascular channels that formed at either time interval, but many of the GFP-expressing cells also expressed Mac 3, CD68, and VEGFR2. Approximately twice as many circulating CD34+/VEGFR2+ cells were found by day 3 in animals with thrombus compared with sham controls (CD45-, P=0.046 and CD45(+), P=0.016). CONCLUSIONS: Bone marrow-derived, Tie2-expressing cells were recruited into the thrombus during resolution but did not line the new vessels. Many of these cells expressed a macrophage phenotype and may represent a population of plastic stem cells that orchestrate thrombus recanalization. PMID- 15883208 TI - Impaired vascular growth in late adolescence after intrauterine growth restriction. AB - BACKGROUND: Abnormal blood flow in a fetus small for gestational age indicates true fetal intrauterine growth restriction (IUGR). We tested the hypothesis that IUGR with abnormal fetal blood flow is associated with long-term abnormal vascular morphology and function in adolescence. METHODS AND RESULTS: In a prospective study, vascular mechanical properties of the common carotid artery (CCA), abdominal aorta, and popliteal artery (PA) were assessed by echo-tracking sonography in 21 adolescents with IUGR and abnormal fetal aortic blood flow and in 23 adolescents with normal fetal growth and normal fetal aortic blood flow. Endothelium-dependent and -independent vasodilatation of the brachial artery was measured by high-resolution ultrasound. After adjustment for body surface area and sex, the IUGR group had significantly smaller end-diastolic vessel diameters than the referents in the abdominal aorta and PA (mean difference, 1.7 mm [95% CI, 0.62 to 2.74] and 0.6 mm [95% CI, 0.25 to 1.02], respectively) (P=0.003 and P=0.002, respectively), with a similar trend in the CCA (P=0.09). A higher resting heart rate was observed in the IUGR group (P=0.01). No differences were found in stiffness or in endothelium-dependent and -independent vasodilatation between the 2 groups. CONCLUSIONS: IUGR caused by placental insufficiency appears to be associated with impaired vascular growth persisting into young adulthood in both men and women. The smaller aortic dimensions and the higher resting heart rate seen in adolescents with previous IUGR may be of importance for future cardiovascular health. PMID- 15883209 TI - Randomized comparison of a titanium-nitride-oxide-coated stent with a stainless steel stent for coronary revascularization: the TiNOX trial. AB - BACKGROUND: Stent coating with titanium-nitride-oxide has been shown to reduce neointimal hyperplasia in the porcine restenosis model. We designed a prospective, randomized, clinical study to investigate the safety and efficacy of titanium-nitride-oxide-coated stents compared with stainless steel stents. METHODS AND RESULTS: Ninety-two patients with de novo lesions were randomly assigned to treatment with titanium-nitride-oxide-coated stents (n=45) or stainless steel stents of otherwise identical design (n=47; control). Baseline characteristics were similar in both groups. At 30 days, no stent thromboses or other adverse events had occurred in either group. Quantitative coronary angiography at 6 months revealed lower late loss (0.55+/-0.63 versus 0.90+/-0.76 mm, P=0.03) and percent diameter stenosis (26+/-17% versus 36+/-24%, P=0.04) in lesions treated with titanium-nitride oxide-coated than in control stents. Binary restenosis was reduced from 33% in the control group to 15% in the titanium nitride oxide-coated stent group (P=0.07). Intravascular ultrasound studies at 6 months showed smaller neointimal volume in titanium-nitride-oxide-coated stents than in control stents (18+/-21 versus 48+/-28 mm3, P<0.0001). Major adverse cardiac events at 6 months were less frequent in titanium-nitride-oxide-coated stents than in control stent-treated patients (7% versus 27%, P=0.02), largely driven by a reduced need for target-lesion revascularization (7% versus 23%, P=0.07). CONCLUSIONS: Revascularization with titanium-nitride-oxide-coated stents is safe and effective in patients with de novo native coronary artery lesions. Titanium-nitride-oxide-coated stents reduce restenosis and major adverse cardiac events compared with stainless steel stents of otherwise identical design. PMID- 15883210 TI - Effect of difficulty affording health care on health status after coronary revascularization. AB - BACKGROUND: An objective of the United States' Healthy People 2010 Initiative is to eliminate disparities based on socioeconomic status. We assessed the effect of difficulty affording health care on the health status (symptoms, function, and quality of life) of patients treated with percutaneous coronary intervention or CABG. METHODS AND RESULTS: A consecutive, single-center cohort of 480 patients undergoing coronary revascularization received the Seattle Angina Questionnaire at the time of their procedure and at subsequent monthly intervals for 6 months. At baseline, patients who reported somewhat of a burden to a severe burden in affording health care had significantly lower scores on the Seattle Angina Questionnaire (mean+/-SD) with respect to angina (55+/-29 versus 68+/-25, P<0.0001), physical limitation (55+/-26 versus 72+/-24, P<0.0001), and quality of life (46+/-22 versus 56+/-22, P<0.0001) than those who did not perceive healthcare costs to be burdensome. Although both groups of patients improved after revascularization, poorer health status persisted among those with difficulty affording health care after percutaneous coronary intervention (6 month mean+/-SE: angina 79+/-2.5 versus 88+/-1.9, P=0.002; physical function 61+/ 2.7 versus 80+/-2.0, P<0.0001; quality of life 67+/-2.4 versus 82+/-1.8, P<0.0001) but not after CABG (angina 91+/-2.5 versus 93+/-1.6, P=0.47; physical function 75+/-3.4 versus 81+/-2.2, P=0.13; quality of life 84+/-3.1 versus 84+/ 2.0, P=0.81). Similar differences remained after adjustment for demographic and clinical characteristics. CONCLUSIONS: Patients reporting difficulty affording health care have worse health status at the time of coronary revascularization. A persistent disparity exists after percutaneous but not surgical revascularization. Additional inquiry into the mechanism of this disparity is needed so that the goals of equitable health care, irrespective of treatment strategy, can be achieved. PMID- 15883211 TI - Regeneration of infarcted myocardium by intramyocardial implantation of ex vivo transforming growth factor-beta-preprogrammed bone marrow stem cells. AB - BACKGROUND: Recent studies have shown that bone marrow-derived stem cells differentiate into the phenotype of cardiomyocytes in vivo and in vitro. We tried to regenerate infarcted myocardium by implanting ex vivo transforming growth factor (TGF)-beta-preprogrammed CD117 (c-kit)-positive (CD117+) stem cells intramyocardially. METHODS AND RESULTS: CD117+ cells were isolated from the bone marrow mononuclear cells of GFP-transgenic or normal C57/BL6 mice. The myogenic differentiation of CD117+ cells was achieved by cultivation with TGF-beta. Using an acute myocardial infarction model, we also tried to regenerate infarcted myocardium by implanting untreated (newly isolated) or preprogrammed (24 hours of cultivation with 5 ng/mL TGF-beta1) CD117+ cells intramyocardially. TGF-beta increased the cellular expression of myosin, troponins, connexin-43, GATA-4, and NKx-2.5, which suggested that it induced the myogenic differentiation of CD117+ cells. Compared with the effects of PBS injection only, the microvessel density in the infarcted myocardium was increased significantly 3 months after the implantation of either TGF-beta-preprogrammed or untreated CD117+ cells. Moreover, many of the TGF-beta-preprogrammed CD117+ cells were stained positively for myosin, whereas few of the untreated CD117+ cells were. Histological analysis revealed newly regenerated myocardium in the left ventricular anterior wall after the implantation of TGF-beta-preprogrammed cells but not untreated cells. Furthermore, the left ventricular percent fraction shortening was significantly higher after the implantation of TGF-beta-preprogrammed cells than after the implantation of untreated CD117+ cells. CONCLUSIONS: TGF-beta conducted the myogenic differentiation of CD117+ stem cells by upregulating GATA-4 and NKx-2.5 expression. Therefore, the intramyocardial implantation of TGF-beta-preprogrammed CD117+ cells effectively assisted the myocardial regeneration and induced therapeutic angiogenesis, contributing to functional cardiac regeneration. PMID- 15883212 TI - Reduced inotropic reserve and increased susceptibility to cardiac ischemia/reperfusion injury in phosphocreatine-deficient guanidinoacetate-N methyltransferase-knockout mice. AB - BACKGROUND: The role of the creatine kinase (CK)/phosphocreatine (PCr) energy buffer and transport system in heart remains unclear. Guanidinoacetate-N methyltransferase-knockout (GAMT-/-) mice represent a new model of profoundly altered cardiac energetics, showing undetectable levels of PCr and creatine and accumulation of the precursor (phospho-)guanidinoacetate (P-GA). To characterize the role of a substantially impaired CK/PCr system in heart, we studied the cardiac phenotype of wild-type (WT) and GAMT-/- mice. METHODS AND RESULTS: GAMT-/ mice did not show cardiac hypertrophy (myocyte cross-sectional areas, hypertrophy markers atrial natriuretic factor and beta-myosin heavy chain). Systolic and diastolic function, measured invasively (left ventricular conductance catheter) and noninvasively (MRI), were similar for WT and GAMT-/- mice. However, during inotropic stimulation with dobutamine, preload-recruitable stroke work failed to reach maximal levels of performance in GAMT-/- hearts (101+/-8 mm Hg in WT versus 59+/-7 mm Hg in GAMT-/-; P<0.05). (31)P-MR spectroscopy experiments showed that during inotropic stimulation, isolated WT hearts utilized PCr, whereas isolated GAMT-/- hearts utilized P-GA. During ischemia/reperfusion, GAMT-/- hearts showed markedly impaired recovery of systolic (24% versus 53% rate pressure product recovery; P<0.05) and diastolic function (eg, left ventricular end-diastolic pressure 23+/-9 in WT and 51+/-5 mm Hg in GAMT-/- during reperfusion; P<0.05) and incomplete resynthesis of P-GA. CONCLUSIONS: GAMT-/- mice do not develop hypertrophy and show normal cardiac function at low workload, suggesting that a fully functional CK/PCr system is not essential under resting conditions. However, when acutely stressed by inotropic stimulation or ischemia/reperfusion, GAMT-/- mice exhibit a markedly abnormal phenotype, demonstrating that an intact, high-capacity CK/PCr system is required for situations of increased cardiac work or acute stress. PMID- 15883213 TI - Angiotensin II activates the Smad pathway in vascular smooth muscle cells by a transforming growth factor-beta-independent mechanism. AB - BACKGROUND: Angiotensin II (Ang II) participates in vascular fibrosis. Transforming growth factor-beta (TGF-beta) is considered the most important fibrotic factor, and Smad proteins are essential components of the TGF-beta signaling system. Our aim was to investigate whether Ang II activates the Smad pathway in vascular cells and its potential role in fibrosis, evaluating connective tissue growth factor (CTGF) and extracellular matrix (ECM) proteins. METHODS AND RESULTS: Systemic infusion of Ang II into Wistar rats increased aortic Smad2, phosphorylated-Smad2, and Smad4 expression, associated with CTGF upregulation. In growth-arrested vascular smooth muscle cells, Ang II treatment for 20 minutes caused Smad2 phosphorylation, nuclear translocation of phosphorylated-Smad2 and Smad4, and increased Smad DNA-binding activity. Ang II also caused Smad overexpression and Smad-dependent gene transcription. The AT1 antagonist losartan diminished Ang II-induced Smad activation. The blockade of endogenous TGF-beta did not modify the activation of Smad caused by Ang II. The p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 diminished Ang II induced Smad2 phosphorylation. These data show that Ang II activates the Smad pathway via AT1 receptors and MAPK activation independently of TGF-beta. Transient transfection with Smad7, which interferes with receptor-mediated activation of Smad2, diminished Ang II-induced CTGF promoter activation, gene and protein expression, and fibronectin and type-1 procollagen overexpression, showing that Smad activation is involved in Ang II-induced fibrosis. CONCLUSIONS: Our results show that Ang II activates the Smad signaling system in vascular cells in vivo and in vitro. Smad proteins are involved in Ang II-induced CTGF and ECM overexpression independently of TGF-beta. This novel finding suggests that Smad activation could be involved in the profibrogenic effects of Ang II in vascular diseases. PMID- 15883214 TI - Dysregulated osteoprotegerin/RANK ligand/RANK axis in clinical and experimental heart failure. AB - BACKGROUND: Persistent inflammation appears to play a role in the development of heart failure (HF). Osteoprotegerin (OPG), the receptor activator of nuclear factor-kappaB (RANK), and RANK ligand (RANKL) are newly discovered members of the tumor necrosis factor superfamily that are critical regulators in bone metabolism but appear also to be involved in immune responses. We hypothesized that the OPG/RANK/RANKL axis could be involved in the pathogenesis of heart failure (HF), and this hypothesis was investigated in both experimental and clinical studies. METHODS AND RESULTS: Our main and novel findings were as follows: (1) In a rat model of postinfarction HF, we found persistently increased gene expression of OPG, RANK, and RANKL in the ischemic part of the left ventricle (LV) and, for OPG, in the nonischemic part that involved both noncardiomyocyte and in particular cardiomyocyte tissue. (2) Enhanced myocardial protein levels of OPG, RANK, and RANKL, in particular, were also seen in human HF, and using immunohistochemistry, we localized these mediators to cardiomyocytes within the LV in both experimental and clinical HF. (3) In human HF, we also found increased systemic expression of RANKL (T cells and serum) and OPG (serum), with increasing levels according to functional, hemodynamic, and neurohormonal disease severity. (4) RANKL increased total matrix metalloproteinase activity in human fibroblasts, which indicates a matrix-degrading net effect and suggests a potential mechanism by which enhanced RANKL expression in HF may contribute to LV dysfunction. CONCLUSIONS: These findings suggest a potential role for known mediators of bone homeostasis in the pathogenesis of HF and possibly represents new targets for therapeutic intervention in this disorder. PMID- 15883215 TI - Pioglitazone decreases carotid intima-media thickness independently of glycemic control in patients with type 2 diabetes mellitus: results from a controlled randomized study. AB - BACKGROUND: Patients with type 2 diabetes mellitus are at high risk of cardiovascular disease. Carotid intima-media thickness (IMT) is a strong predictor of myocardial infarction and stroke. METHODS AND RESULTS: We compared the effects of pioglitazone-based therapy (45 mg/d) and glimepiride-based treatment (2.7+/-1.6 mg/d) for 12 and 24 weeks on metabolic control (HbA1c), insulin resistance (homeostasis model assessment), and carotid IMT (B-mode ultrasonography) in a randomized controlled study in 173 orally treated patients with type 2 diabetes (66 women, 107 men; mean+/-SD age, 62.6+/-7.9 years; body mass index, 31.8+/-4.6 kg/m2; HbA1c, 7.5+/-0.9%). Treatment was generally well tolerated in both groups. Despite similar improvements in metabolic control (HbA1c) after 24 weeks (-0.8+/-0.9% [pioglitazone] versus -0.6+/-0.8% [glimepiride]; P=NS), carotid IMT was reduced only in the pioglitazone group after 12 weeks (-0.033+/-0.052 versus -0.002+/-0.047 mm [glimepiride]; P<0.01 between groups) and 24 weeks (-0.054+/-0.059 versus -0.011+/-0.058 mm [glimepiride]; P<0.005 between groups). Insulin resistance was also improved only in the pioglitazone group (homeostasis model assessment, -2.2+/-3.4 versus -0.3+/ 3.3; P<0.0001 between groups). Reduction of IMT correlated with improvement in insulin resistance (r=0.29, P<0.0005) and was independent of improvement in glycemic control (r=0.03, P=0.68). CONCLUSIONS: We found a substantial regression of carotid IMT, independent of improved glycemic control, after 12 and 24 weeks of pioglitazone treatment. This finding may have important prognostic implications for patients with type 2 diabetes mellitus. PMID- 15883216 TI - Stimulation of paracrine pathways with growth factors enhances embryonic stem cell engraftment and host-specific differentiation in the heart after ischemic myocardial injury. AB - BACKGROUND: Growth factors play an essential role in organogenesis. We examine the potential of growth factors to enhance cell engraftment and differentiation and to promote functional improvement after transfer of undifferentiated embryonic stem cells into the injured heart. METHODS AND RESULTS: Green fluorescent protein (GFP)-positive embryonic stem cells derived from 129sv mice were injected into the ischemic area after left anterior descending artery ligation in allogenic (BALB/c) mice. Fifty nanograms of recombinant mouse vascular endothelial growth factor, fibroblast growth factor (FGF), and transforming growth factor (TGF) was added to the cell suspension. Separate control groups were formed in which only the growth factors were given. Echocardiography was performed 2 weeks later to evaluate heart function (fractional shortening [FS]), end-diastolic diameter, and left ventricular wall thickness). Hearts were harvested for histology (connexin 43, alpha-sarcomeric actin, CD3, CD11c, major histocompatability complex class I, hematoxylin-eosin). Degree of restoration (GFP-positive graft/infarct area ratio), expression of cardiac markers, host response, and tumorigenicity were evaluated. Cell transfer resulted in improved cardiac function. TGF-beta led to better restorative effect and a stronger expression of connexin 43, alpha-sarcomeric actin, and major histocompatability complex class I. TGF-beta and FGF retained left ventricular diameter. FS was better in the TGF-beta, FGF, and embryonic stem cells-only group compared with left anterior descending artery-ligated controls. Growth factors with cells (TGF-beta, FGF) resulted in higher FS and smaller end-diastolic diameter than growth factors alone. CONCLUSIONS: Growth factors can promote in vivo organ-specific differentiation of early embryonic stem cells and improve myocardial function after cell transfer into an area of ischemic lesion. TGF-beta should be considered as an adjuvant for myocardial restoration with the use of embryonic stem cells. PMID- 15883217 TI - Increased prevalence of coronary artery aneurysms among cocaine users. AB - BACKGROUND: Cocaine abuse has been implicated in multiple cardiovascular complications. Coronary artery aneurysms (CAAs) and ectasia occur in 0.2% to 5.3% of patients referred for angiography and are associated with atherosclerosis, Kawasaki's disease, and several rare disorders. After observing CAAs in multiple young cocaine users, we investigated the prevalence of CAAs among cocaine users undergoing coronary angiography. METHODS AND RESULTS: Clinical and angiographic characteristics of 112 consecutive patients with a history of cocaine use and coronary angiography were compared with a control group of similar age and risk factors from an existing angiographic database over the same time period. Coronary angiograms were independently read by 3 reviewers blinded to cocaine use. Cocaine users were young (mean age, 44 years), predominantly male (80%), and cigarette smokers (95%). Control patients had higher rates of diabetes (33%) and more severe coronary artery disease (P=0.01). Previous myocardial infarction was common in both groups (45% of cocaine users, 38% of control patients). Despite the frequent history of myocardial infarction among cocaine users, 48% had nonobstructive coronary artery disease. Among cocaine users, 34 of 112 (30.4%) had CAAs compared with 6 of 79 (7.6%) in the control group (P<0.001). Cocaine use was a strong predictor of CAA by univariate and multivariate analyses. CONCLUSIONS: This is the first description of an association between cocaine use and CAA. The prevalence of CAA among cocaine users was higher than expected (30.4%), given such a young cohort. Cocaine use may predispose to the formation of CAA, which may in turn be a contributing factor to myocardial infarction. PMID- 15883218 TI - Images in cardiovascular medicine. Leaflet fracture of a St. Jude mechanical bileaflet valve. PMID- 15883219 TI - Images in cardiovascular medicine. Fast and complete healing of 2 coronary artery spontaneous dissections. PMID- 15883220 TI - Letter regarding article by Tsimikas et al, "high-dose atorvastatin reduces total plasma levels of oxidized phospholipids and immune complexes present on apolipoprotein B-100 in patients with acute coronary syndromes in the MIRACL trial". PMID- 15883221 TI - Letter regarding article by Selvanayagam et al, "value of delayed-enhancement cardiovascular magnetic resonance imaging in predicting myocardial viability after surgical revascularization". PMID- 15883222 TI - Letter regarding article by Jaeggi et al, "transplacental fetal treatment improves the outcome of prenatally diagnosed complete atrioventricular block without structural heart disease". PMID- 15883223 TI - On mice, rabbits, and human heart failure. PMID- 15883224 TI - Clinical significance of statin pleiotropic effects: hypotheses versus evidence. PMID- 15883225 TI - B-type natriuretic peptide in organic mitral regurgitation: determinants and impact on outcome. AB - BACKGROUND: B-type natriuretic peptide (BNP) activation observed in cardiac diseases is a predictor of poor outcome; however, in organic mitral regurgitation (MR), BNP determinants and prognostic value are unknown. METHODS AND RESULTS: We prospectively enrolled 124 patients with chronic organic MR (aged 63+/-15 years, 60% males) in whom we measured BNP level and simultaneously quantified MR degree, left ventricular (LV) remodeling, and left atrial (LA) volumes and analyzed long term outcome. Baseline BNP level (54+/-67 pg/mL, median 31 pg/mL) was associated univariately with multiple clinical and echocardiographic characteristics, but in multivariate analysis, independent determinants of BNP, beyond age and sex (both P< or =0.01), were LV end-systolic volume index, LA volume, atrial fibrillation, and symptoms (all P<0.02). Conversely, MR degree was not independently associated with BNP. During follow-up, patients with high versus low BNP (> or =31 versus <31 pg/mL) displayed lower survival rates (at 5 years, 72+/-10% versus 95+/-5%, P=0.03) and higher rates of the combined end point of death and heart failure (at 5 years, 42+/-10% versus 16+/-7%, P=0.03). In multivariate analysis, with adjustment for age, sex, functional class, MR severity, and ejection fraction, BNP was independently predictive of mortality (hazard ratio per 10 pg/mL, 1.23 [95% CI 1.07 to 1.48], P=0.004) and of death or heart failure (hazard ratio per 10 pg/mL, 1.09 [95% CI 1.001 to 1.19], P=0.04). CONCLUSIONS: BNP activation in organic MR reflects primarily ventricular and atrial consequences rather than degree of MR. Higher BNP level in patients with organic MR independently predicts adverse events under conservative management. Therefore, BNP activation in organic MR is an emerging biomarker of severity of MR consequences and of poor clinical outcome, and its assessment should be considered in the clinical evaluation and risk stratification of patients with MR. PMID- 15883226 TI - Chronic venous insufficiency. PMID- 15883227 TI - Ouabain and serum sodium. PMID- 15883229 TI - Vascular and metabolic effects of combined therapy with ramipril and simvastatin in patients with type 2 diabetes. AB - Mechanisms underlying biological effects of statin and angiotensin-converting enzyme inhibitor therapies differ. Therefore, we compared vascular and metabolic responses to these therapies either alone or in combination in patients with type 2 diabetes. This was a randomized, double-blind, placebo-controlled crossover trial with 3 treatment arms (each 2 months) and 2 washout periods (each 2 months). Fifty patients with type 2 diabetes were given simvastatin 20 mg and placebo, simvastatin 20 mg and ramipril 10 mg, or ramipril 10 mg and placebo daily during each 2-month treatment period. Ramipril alone or combined therapy significantly reduced blood pressure when compared with simvastatin alone. When compared with ramipril alone, simvastatin alone or combined therapy significantly improved the lipoprotein profile. All 3 treatment arms significantly improved flow-mediated dilator response to hyperemia and reduced plasma levels of malondialdehyde relative to baseline measurements. However, these parameters were changed to a greater extent with combined therapy when compared with simvastatin or ramipril alone (P<0.001 by ANOVA). When compared with simvastatin or ramipril alone, combined therapy significantly reduced high-sensitivity C-reactive protein levels (P=0.004 by ANOVA). Interestingly, combined therapy or ramipril alone significantly increased plasma adiponectin levels and insulin sensitivity relative to baseline measurements. These changes were significantly greater than in the group treated with simvastatin alone (P<0.015 by ANOVA). Ramipril combined with simvastatin had beneficial vascular and metabolic effects when compared with monotherapy in patients with type 2 diabetes. PMID- 15883230 TI - Cyclooxygenase-1-deficient mice have high sleep-to-wake blood pressure ratios and renal vasoconstriction. AB - We used cyclooxygenase-1 (COX-1)-deficient mice to test the hypothesis that COX-1 regulates blood pressure (BP) and renal hemodynamics. The awake time (AT) mean arterial pressures (MAPs) measured by telemetry were not different between COX 1(+/+) and COX-1(-/-) (131+/-2 versus 126+/-3 mm Hg; NS). However, COX-1(-/-) had higher sleep time (ST) MAP (93+/-1 versus 97+/-2 mm Hg; P<0.05) and sleep-to awake BP ratio (+8.6%; P<0.05). Under anesthesia with moderate sodium loading, COX-1(-/-) had higher MAP (109+/-5 versus 124+/-4 mm Hg; P<0.05), renal vascular resistance (23.5+/-1.6 versus 30.7+/-1.7 mm Hg . mL(-1) . min(-1) . g(-1); P<0.05) and filtration fraction (33.7+/-2.1 versus 40.2+/-2.0%; P<0.05). COX-1(-/ ) had a 89% reduction (P<0.0001) in the excretion of TxB2, a 76% reduction (P<0.01) in PGE2, a 40% reduction (P<0.0002) in 6-ketoPGF1alpha (6keto), a 27% reduction (P<0.02) in 11-betaPGF2alpha (11beta), a 35% reduction (P<0.01) in nitrate plus nitrite (NOx), and a 52% increase in metanephrine (P<0.02). The excretion of normetanephrine, a marker for sympathetic nervous activity, was reduced during ST in COX-1(+/+) (6.9+/-0.9 versus 3.2+/-0.6 g . g(-1) creatinine . 10(-3); P<0.01). This was blunted in COX-1(-/-) (5.1+/-0.9 versus 4.9+/-0.7 g . g(-1) creatinine . 10(-3); NS). Urine collection during ST showed lower excretion of 6keto, 11beta, NOx, aldosterone, sodium, and potassium than during AT in both COX-1(+/+) and COX-1(-/-), and there were positive correlations among these parameters (6keto versus NOx; P<0.005; 11beta versus NOx; P<0.005; and NOx versus sodium; P<0.005). In conclusion, COX-1 mediates a suppressed sympathetic nervous activity and enhanced NO, which may contribute to renal vasodilatation and a reduced MAP while asleep or under anesthesia. COX-1 contributes to the normal nocturnal BP dipping phenomenon. PMID- 15883231 TI - Loss of vascular regulation by soluble guanylate cyclase is emerging as a key target of the hypertensive disease process. PMID- 15883232 TI - Human-antigen R (HuR) expression in hypertension: downregulation of the mRNA stabilizing protein HuR in genetic hypertension. AB - In aged spontaneously hypertensive rats (SHR), vasorelaxant responses to NO are attenuated compared with normotensive control rats (Wistar-Kyoto [WKY]) because of a decreased expression of the important NO receptor soluble guanylyl cyclase (sGC). Because the expression of sGC subunits alpha1 and beta1 is controlled at the post-transcriptional level by the mRNA-binding protein human-antigen R (HuR), we now assessed whether or not altered expression of HuR could account for downregulation of sGCalpha1 and sGCbeta1 in genetic hypertension. The expression of HuR (and sGCalpha1 and sGCbeta1) in aortas from aged SHR was significantly decreased at the mRNA and protein level compared with age-matched WKY rats, whereas expression of HuR was not different in prehypertensive young (2 months of age) SHR and age-matched WKY rats. The mRNA-binding activity of HuR in native aortic protein extracts from aged SHR was markedly reduced compared with normotensive WKY rats, as detected by RNA electrophoretic mobility shift analysis, using biotin-labeled adenine and uracil (AU)-rich element (ARE) containing RNA probes from the 3'-untranslated region of sGCalpha1 and sGCbeta1. In contrast, ARE-binding activity was not different between prehypertensive young SHR and young WKY rats. In vitro RNA degradation assays using the same AU-rich sGC mRNA probes revealed an accelerated sGCalpha1 and sGCbeta1 mRNA decay in the presence of native protein extract from hypertensive SHR, which was less rapid with aortic protein from normotensive WKY rats. These findings suggest that in this animal model of genetic hypertension, the reduced expression of sGC subunits is mediated by downregulation of the sGC mRNA-stabilizing protein HuR. PMID- 15883233 TI - Generation of peripheral sensory and sympathetic neurons and neural crest cells from human embryonic stem cells. AB - Human embryonic stem cells (hESCs) have been directed to differentiate into neuronal cells using many cell-culture techniques. Central nervous system cells with clinical importance have been produced from hESCs. To date, however, there have been no definitive reports of generation of peripheral neurons from hESCs. We used a modification of the method of Sasai and colleagues for mouse and primate embryonic stem cells to elicit neuronal differentiation from hESCs. When hESCs are cocultured with the mouse stromal line PA6 for 3 weeks, neurons are induced that coexpress (a) peripherin and Brn3a, and (b) peripherin and tyrosine hydroxylase, combinations characteristic of peripheral sensory and sympathetic neurons, respectively. In vivo, peripheral sensory and sympathetic neurons develop from the neural crest (NC). Analysis of expression of mRNAs identified in other species as NC markers reveals that the PA6 cells induce NC-like cells before neuronal differentiation takes place. Several NC markers, including SNAIL, dHAND, and Sox9, are increased at 1 week of coculture relative to naive cells. Furthermore, the expression of several NC marker genes known to be downregulated upon in vivo differentiation of NC derivatives, was observed to be present at lower levels at 3 weeks of PA6-hESC coculture than at 1 week. Our report is the first on the expression of molecular markers of NC-like cells in primates, in general, and in humans, specifically. Our results suggest that this system can be used for studying molecular and cellular events in the almost inaccessible human NC, as well as for producing normal human peripheral neurons for developing therapies for diseases such as familial dysautonomia. PMID- 15883234 TI - The milk debate. PMID- 15883235 TI - Risk of hospitalization for myocardial infarction among users of rofecoxib, celecoxib, and other NSAIDs: a population-based case-control study. AB - BACKGROUND: It remains uncertain if the excess cardiovascular risk of rofecoxib and celecoxib reported in clinical trials is present in routine practice and whether the use of other nonaspirin nonsteroidal anti-inflammatory drugs (NSAIDs) also carries an increased cardiovascular risk. We performed a population-based case-control study to examine the risk of myocardial infarction (MI) among users of various categories of nonaspirin NSAIDs. METHODS: Using data from hospital discharge registries in the counties of North Jutland, Viborg, and Aarhus, Denmark, and the Danish Civil Registration System, we identified 10,280 cases of first-time hospitalization for MI and 102,797 sex- and age-matched non-MI population controls. All prescriptions for nonaspirin NSAIDs filled before the date of admission for MI were identified using population-based prescription databases. Relative risk estimates for MI were adjusted for a history of cardiovascular disease, hypertension, diabetes mellitus, chronic bronchitis or emphysema, alcoholism, liver cirrhosis, upper gastrointestinal bleeding, rheumatoid arthritis, systemic lupus erythematosus and the use of high-dose aspirin, platelet inhibitors, insulin or oral hypoglycemic drugs, antihypertensive drugs, lipid-lowering drugs, oral anticoagulants, nitrates, penicillamine, gold, oral glucocorticoids, and hormone therapy before the date of admission for MI. RESULTS: Current users of rofecoxib had an elevated risk estimate for hospitalization for MI compared with nonusers of any category of nonaspirin NSAIDs (adjusted relative risk [ARR], 1.80; 95% confidence interval [CI], 1.47-2.21). Increased risk estimates were also found among current users of celecoxib (ARR, 1.25; 95% CI, 0.97-1.62), other cyclooxygenase-2 selective inhibitors (ARR, 1.45; 95% CI, 1.09-1.93), naproxen (ARR, 1.50; 95% CI, 0.99 2.29), and other conventional nonaspirin NSAIDs (ARR, 1.68; 95% CI, 1.52-1.85). The highest ARRs were found among new users of all examined drug categories. CONCLUSIONS: Current and new users of all classes of nonaspirin NSAIDs had elevated relative risk estimates for MI. Although the increased risk estimates may partly reflect unmeasured bias, they indicate the need for further examination of the cardiovascular safety of all nonaspirin NSAIDs. PMID- 15883236 TI - Reduction of alcohol consumption by brief alcohol intervention in primary care: systematic review and meta-analysis. AB - BACKGROUND: Numerous trials of the efficacy of brief alcohol intervention have been conducted in various settings among individuals with a wide range of alcohol disorders. Nevertheless, the efficacy of the intervention is likely to be influenced by the context. We evaluated the evidence of efficacy of brief alcohol interventions aimed at reducing long-term alcohol use and related harm in individuals attending primary care facilities but not seeking help for alcohol related problems. METHODS: We selected randomized trials reporting at least 1 outcome related to alcohol consumption conducted in outpatients who were actively attending primary care centers or seeing providers. Data sources were the Cochrane Central Register of Controlled Trials, MEDLINE, PsycINFO, ISI Web of Science, ETOH database, and bibliographies of retrieved references and previous reviews. Study selection and data abstraction were performed independently and in duplicate. We assessed the validity of the studies and performed a meta-analysis of studies reporting alcohol consumption at 6 or 12 months of follow-up. RESULTS: We examined 19 trials that included 5639 individuals. Seventeen trials reported a measure of alcohol consumption, of which 8 reported a significant effect of intervention. The adjusted intention-to-treat analysis showed a mean pooled difference of -38 g of ethanol (approximately 4 drinks) per week (95% confidence interval, -51 to -24 g/wk) in favor of the brief alcohol intervention group. Evidence of other outcome measures was inconclusive. CONCLUSION: Focusing on patients in primary care, our systematic review and meta-analysis indicated that brief alcohol intervention is effective in reducing alcohol consumption at 6 and 12 months. PMID- 15883237 TI - Dairy consumption and risk of type 2 diabetes mellitus in men: a prospective study. AB - BACKGROUND: Diet and lifestyle modifications can substantially reduce the risk of type 2 diabetes. While a strong inverse association has been reported between dairy consumption and the insulin resistance syndrome among young obese adults, the relation between dairy intake and type 2 diabetes is unknown. METHODS: We prospectively examined the relation between dairy intake and incident cases of type 2 diabetes in 41,254 male participants with no history of diabetes, cardiovascular disease, and cancer at baseline in the Health Professionals Follow up Study. RESULTS: During 12 years of follow-up, we documented 1243 incident cases of type 2 diabetes. Dairy intake was associated with a modestly lower risk of type 2 diabetes. After adjusting for potential confounders, including body mass index, physical activity, and dietary factors, the relative risk for type 2 diabetes in men in the top quintile of dairy intake was 0.77 (95% confidence interval [CI], 0.62-0.95; P for trend, .003) compared with those in the lowest quintile. Each serving-per-day increase in total dairy intake was associated with a 9% lower risk for type 2 diabetes (multivariate relative risk, 0.91; 95% CI, 0.85-0.97). The corresponding relative risk was 0.88 (95% CI, 0.81-0.94) for low fat dairy intake and 0.99 (95% CI, 0.91-1.07) for high-fat dairy intake. The association did not vary significantly according to body mass index (< 25 vs > or = 25 kg/m(2); P for interaction, .57). CONCLUSION: Dietary patterns characterized by higher dairy intake, especially low-fat dairy intake, may lower the risk of type 2 diabetes in men. PMID- 15883238 TI - A prospective study of physical activity and incident and fatal prostate cancer. AB - BACKGROUND: Whether physical activity has benefits against prostate cancer incidence or progression is unclear. Therefore, we assessed physical activity in relation to prostate cancer incidence, mortality, and Gleason histologic grade. METHODS: We used data from the Health Professionals Follow-up Study, a prospective cohort study, to determine the number of cases of incident, advanced (seminal vesicle invasion, metastasis, or fatal), fatal, and high-grade prostate cancer in a cohort of 47,620 US male health professionals, followed up from February 1, 1986, to January 31, 2000. RESULTS: During 14 years of follow-up, we documented 2892 new cases of prostate cancer, including 482 advanced cases (280 of which were fatal). For total prostate cancer, no association was observed for total, vigorous, and nonvigorous physical activity. In men 65 years or older, we observed a lower risk in the highest category of vigorous activity for advanced (multivariable relative risk, 0.33; 95% confidence interval, 0.17-0.62, for more than 29 vs 0 metabolic equivalent hours) and for fatal (relative risk, 0.26; 95% confidence interval, 0.11-0.66) prostate cancer. No associations were observed in younger men. Differential screening by prostate-specific antigen or a reduction in physical activity due to undiagnosed prostate cancer did not appear to account for the results. Among cases, men with high levels of physical activity were less likely to be diagnosed with poorly differentiated cancers (Gleason grade > or = 7). CONCLUSION: Although the mechanisms are not yet understood, these findings suggest that regular vigorous activity could slow the progression of prostate cancer and might be recommended to reduce mortality from prostate cancer, particularly given the many other documented benefits of exercise. PMID- 15883239 TI - Long-term intake of trans-fatty acids and risk of gallstone disease in men. AB - BACKGROUND: The consumption of trans-fatty acids adversely affects blood lipid levels. The relationship with the incidence of gallstone disease is unknown. METHODS: We prospectively studied consumption of trans-fatty acids in relation to the risk of gallstone disease in a cohort of 45,912 men. trans-Fatty acid consumption was assessed using a validated semiquantitative food frequency questionnaire. Newly diagnosed gallstone disease, by radiology or cholecystectomy, was ascertained biennially. RESULTS: During 14 years of follow up, we documented 2356 new cases of symptomatic gallstones. After adjusting for age and other potential risk factors, we found that compared with men in the lowest quintile of dietary intake of trans-fatty acids, the relative risk (RR) of gallstone disease for those in the highest quintile was 1.23 (95% confidence interval [CI], 1.04-1.44; P for trend, .03). Among individual trans-fatty acids, the RR for trans-oleic fatty acid, when extreme quintiles were compared, was 1.24 (95% CI, 1.06-1.45; P for trend, .02). Intakes of trans-palmitoleic fatty acid (RR, 1.09; 95% CI, 0.90-1.31), trans,trans 18:2 fatty acid (RR, 1.14; 95% CI, 0.96-1.34), and cis-trans 18:2 fatty acid (RR, 1.00; 95% CI, 0.86-1.16) were not significantly associated with the risk. CONCLUSIONS: Our results suggest that a higher intake of trans-fatty acids modestly increases risk of gallstone disease. This adds to the concern that partial hydrogenation of vegetable oils to form shortening and margarine can lead to adverse health effects. PMID- 15883240 TI - Adverse health behaviors and the detection of partner violence by clinicians. AB - BACKGROUND: Intimate partner violence (IPV) is a major public health problem in the United States, and victims are commonly encountered in medical settings. Many barriers exist to clinician-initiated screening for IPV. However, smoking and problem drinking are conditions that clinicians commonly screen for and both have been strongly associated with IPV in prior studies. By estimating the predicted probability of 12-month and lifetime IPV for a given patient based on whether she presents with these conditions, our study gives clinicians information that can help them identify patients at risk for IPV. METHODS: A cross-sectional written patient survey was administered to 2386 female patients at 8 different health care settings in the Greater Boston (Mass) metropolitan area. The probabilities of 12-month and lifetime IPV were estimated based on the women's self-report of smoking and drinking behaviors. RESULTS: A woman who neither smoked nor engaged in problem drinking had a 10% probability of IPV in the preceding 12 months and a 39% chance of IPV in her lifetime. Smoking increased the probability to 14% and 49%, respectively. Problem drinking resulted in a doubling of the predicted probability of 12-month IPV to 21%, with a lifetime probability of 43%. When both conditions were present, the effects were additive, with a woman having a 27% probability of experiencing IPV in the preceding 12 months and 54% chance of IPV in her lifetime. CONCLUSIONS: The presence of smoking or problem drinking should raise clinicians' suspicion for IPV. This paradigm should not replace direct questioning about IPV but may aid in the detection of abuse in patient populations. PMID- 15883241 TI - Marital quality and occurrence of the metabolic syndrome in women. AB - BACKGROUND: Marital status is associated with the early stages and progression of cardiovascular disease, an association that may stem in part from the influence of marital quality on metabolic factors. The objective of this study was to examine whether women reporting marital satisfaction are at reduced risk of developing the metabolic syndrome compared with other women. METHODS: Four hundred thirteen middle-aged women from the Pittsburgh Healthy Women Study completed measures of marital status and marital satisfaction at baseline and 3 years later. Metabolic syndrome (diagnosed according to the criteria of the National Cholesterol Education Program) was assessed at baseline and at the last follow-up visit (an average of 11.5 years later). RESULTS: Compared with maritally satisfied women, maritally dissatisfied (odds ratio [OR], 3.02; 95% confidence interval [CI], 1.46-6.24), divorced (OR, 2.47; 95% CI, 1.02-5.97), and widowed (OR, 5.82; 95% CI, 1.88-18.03) women were significantly more likely to have the metabolic syndrome at follow-up. The differences between maritally satisfied women and dissatisfied (OR, 3.18; 95% CI, 1.42-7.15) and widowed (OR, 5.69; 95% CI, 1.70-9.04) women remained significant in the full multivariate model. The difference between maritally satisfied women and divorced women (OR, 2.35; 95% CI, 0.89-6.18) was reduced to marginal significance in the full multivariate model. Single (OR, 2.84; 95% CI, 0.84-9.64) and moderately satisfied (OR, 1.06; 95% CI, 0.35-3.21) women did not differ significantly from maritally satisfied women. CONCLUSIONS: Women in high-quality marriages are at lower risk of developing the metabolic syndrome. Social histories of patients should include assessment of marital quality. PMID- 15883242 TI - Cardiovascular risk profile earlier in life and Medicare costs in the last year of life. AB - BACKGROUND: Health care costs are generally highest in the year before death, and much attention has been directed toward reducing costs for end-of-life care. However, it is unknown whether cardiovascular risk profile earlier in life influences health care costs in the last year of life. This study addresses this question. METHODS: Prospective cohort of adults from the Chicago Heart Association Detection Project in Industry included 6582 participants (40% women), aged 33 to 64 years at baseline examination (1967-1973), who died at ages 66 to 99 years. Medicare billing records (1984-2002) were used to obtain cardiovascular disease-related and total charges (adjusted to year 2002 dollars) for inpatient and outpatient services during the last year of life. Participants were classified as having favorable levels of all major cardiovascular risk factors (low risk), that is, serum cholesterol level lower than 200 mg/dL (<5.2 mmol/L), blood pressure 120/80 mm Hg or lower and no antihypertensive medication, body mass index (calculated as weight in kilograms divided by the square of height in meters) lower than 25, no current smoking, no diabetes, and no electrocardiographic abnormalities, or unfavorable levels of any 1 only, any 2 only, any 3 only, or 4 or more of these risk factors. RESULTS: In the last year of life, average Medicare charges were lowest for low-risk persons. For example, cardiovascular disease-related and total charges were lower by 10,367 dollars and 15,318 dollars compared with those with 4 or more unfavorable risk factors; the fewer the unfavorable risk factors, the lower the Medicare charges (P for trends <.001). Analyses by sex showed similar patterns. CONCLUSION: Favorable cardiovascular risk profile earlier in life is associated with lower Medicare charges at the end of life. PMID- 15883243 TI - Reasons for intentional weight loss, unintentional weight loss, and mortality in older men. AB - BACKGROUND: We have examined the relationship between intentional and unintentional weight loss and the reasons underlying intention to lose weight and all-cause mortality and mortality due to cardiovascular disease (CVD) and non-CVD causes in older men. METHODS: Prospective study of 4869 men aged 56 to 75 years drawn from general practices in 24 British towns, who in 1996 completed questionnaires about intentional and unintentional weight loss over the preceding 4 years and were followed up for a subsequent 7 years. RESULTS: Unintentional but not intentional weight loss was associated with a significant increase in risk of all-cause mortality compared with men who reported no weight change, even after adjustment for lifestyle characteristics and preexisting disease (adjusted relative risk [RR], 1.71; 95% confidence interval [CI], 1.33-2.19; and RR, 1.00; 95% CI, 0.91-1.10, respectively). Men who lost weight intentionally as a result of personal choice showed significant benefit in all-cause mortality (RR, 0.59; 95% CI, 0.34-1.00; P = .05), which was largely owing to a significant reduction in mortality from non-CVD causes (RR, 0.36; 95% CI, 0.15-0.87). The benefit in these men was most apparent in markedly overweight men (BMI [calculated as weight in kilograms divided by the square of height in meters] > or = 28) and in younger men (age < 65 years). Men who lost weight intentionally owing to ill health or physician's advice showed an increased risk of all-cause mortality (RR, 1.37; 95% CI, 0.96-1.94). No harm or benefit was seen for CVD mortality, irrespective of reasons for intentional weight loss. CONCLUSION: Intentional weight loss carried out for personal reasons is associated with a significant reduction in all-cause mortality in markedly overweight men, and the data suggest that the earlier the intervention, the greater the chance of benefit. PMID- 15883244 TI - Ethnic differences in blood pressure control among men at Veterans Affairs clinics and other health care sites. AB - BACKGROUND: Differential access to health care may contribute to lower blood pressure (BP) control rates to under 140/90 mm Hg in African American compared with white hypertensive patients, especially men (26.5% vs 36.5% of all hypertensive patients in the National Health and Nutrition Examination Survey 1999-2000). The Department of Veterans Affairs (VA) system, which provides access to health care and medications across ethnic and economic boundaries, may reduce disparities in BP control. METHODS: To test this hypothesis, BP treatment and control groups were compared between African American (VA, n = 4379; non-VA, n = 2754) and white (VA, n = 7987; non-VA, n = 4980) hypertensive men. RESULTS: In both groups, whites were older than African Americans (P<.05), had lower BP (P<.001), and had BP controlled to below 140/90 mm Hg more often on their last visit (P<.01). Blood pressure control to below 140/90 mm Hg was comparable among white hypertensive men at VA (55.6%) and non-VA (54.2%) settings (P = .12). In contrast, BP control was higher among African American hypertensive men at VA (49.4%) compared with non-VA (44.0%) settings (P<.01), even after controlling for age, numerous comorbid conditions, and rural-urban classification. African American hypertensive men received a comparable number of prescriptions for BP medications at VA sites (P = .18) and more prescriptions at non-VA sites than did whites (P<.001). African Americans had more visits in the previous year at VA sites (P<.001) and fewer visits at non-VA sites (P<.001) compared with whites. CONCLUSIONS: The ethnic disparity in BP control between African Americans and whites was approximately 40% less at VA than at non-VA health care sites (6.2% vs 10.2%; P<.01). Ensuring access to health care could constitute one constructive component of a national initiative to reduce ethnic disparities in BP control and cardiovascular risk. PMID- 15883245 TI - Alcohol consumption and the risk of renal dysfunction in apparently healthy men. AB - BACKGROUND: Moderate alcohol consumption has been consistently associated with beneficial health effects on cardiovascular disease. In contrast, the association between alcohol consumption and renal dysfunction is less clear. METHODS: We conducted a prospective cohort study of 11,023 initially healthy men who provided blood samples 14 years after a baseline assessment of alcohol consumption. We categorized alcohol consumption into 1 or fewer, 2 to 4, 5 to 6, and 7 or more drinks per week. The main outcome measures were elevated creatinine levels (defined as > or = 1.5 mg/dL [> or = 133 micromol/L]) and reduced estimated glomerular filtration rates (defined as < or = 55 mL/min). We used logistic regression to calculate multivariable-adjusted odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: After 14 years, 473 men (4.3%) had elevated creatinine levels and 1296 (11.8%) had reduced glomerular filtration rates. Compared with men who consumed no more than 1 drink per week, men who consumed 2 to 4 drinks weekly had a multivariable-adjusted OR of 1.04 (95% CI, 0.81-1.32), men who consumed 5 to 6 drinks per week had an OR of 0.92 (95% CI, 0.68-1.25), and men who consumed at least 7 drinks weekly had an OR of 0.71 (95% CI, 0.55 0.92) (P = .01 for trend across categories). Similar associations were observed between alcohol consumption and decreased glomerular filtration rates. Hypertension, diabetes mellitus, and cholesterol level did not attenuate these effects. CONCLUSIONS: In this large cohort of apparently healthy men, alcohol consumption was not associated with an increased risk of renal dysfunction. Instead, these data suggest an inverse relationship between moderate alcohol consumption and the risk of renal dysfunction. PMID- 15883246 TI - Health concerns of patients with nonbacterial prostatitis/pelvic pain. AB - BACKGROUND: Heightened awareness of patients' reasons for physician visits for male pelvic pain (nonbacterial prostatitis) and symptom concerns may increase patient satisfaction with care and help guide better management of this syndrome, for which evidence-based treatment is lacking. METHODS: We interviewed men with recent health maintenance organization visits for new episodes of nonbacterial prostatitis (N = 286; mean age, 46.7 years) and again 3, 6, and 12 months later. We inquired about their reasons for the visit and, at each interview, symptom concerns. We used Poisson regression to examine the association between baseline symptom worry and health care utilization during the 14 months after the index visit. RESULTS: Most patients reported concern at the index visit that they might have an infection (73%) or cancer (68%). One year later, 43% reported prostatitis symptoms in the past month. Among these, many were still concerned that their symptoms would worsen if untreated (71%), that they had cancer (46%) or an infection (45%), and that they might need surgery (44%). Controlling for patient age and baseline symptom severity, we found that baseline symptom worry predicted prostatitis-related health care visits over the 14 months after the index visit (P = .005). CONCLUSIONS: Despite symptom improvement following a health care visit for a new episode of pelvic pain/nonbacterial prostatitis, continued patient concerns about cancer, infection, and symptom worsening without treatment were common, even 1 year later. Patient worry may be associated with increased health care utilization. PMID- 15883247 TI - Retinal arteriolar narrowing, hypertension, and subsequent risk of diabetes mellitus. AB - BACKGROUND: Microvascular disease and hypertension have been linked with risk of diabetes mellitus. We examined the association of retinal arteriolar narrowing, a marker of chronic hypertension, with incident diabetes. METHODS: Prospective cohort study of 3251 nondiabetic persons aged 43 to 86 years living in Wisconsin. The diameters of retinal vessels were measured from baseline retinal photographs of participants. Retinal measurements were summarized as the retinal arteriole-to venule ratio, with smaller ratios indicating narrower arteriolar diameters. Incident diabetes cases were ascertained at the 5-year and 10-year follow-up examinations. RESULTS: There were 249 incident diabetes cases. Participants with narrower retinal arteriolar diameters had a higher incidence of diabetes (cumulative incidences of 5.1%, 7.0%, 9.2%, and 11.7%, comparing decreasing quartiles of arteriole-to-venule ratio). After controlling for baseline casual blood glucose level, glycosylated hemoglobin level, body mass index, and other risk factors, retinal arteriolar narrowing was significantly associated with risk of incident diabetes (multivariable-adjusted relative risk, 1.53; 95% confidence interval, 1.03-2.27; comparing smallest to largest arteriole-to-venule ratio quartiles). Participants with both hypertension and retinal arteriolar narrowing had a 3-fold higher risk of incident diabetes (multivariable-adjusted relative risk, 3.41; 95% confidence interval, 1.66-6.98) than normotensive participants without arteriolar narrowing. CONCLUSIONS: Retinal arteriolar narrowing is related to risk of incident diabetes. These data suggest a possible link between systemic arteriolar narrowing associated with hypertension and diabetes development. PMID- 15883248 TI - Are cyclooxygenase-2 inhibitors being taken only by those who need them? PMID- 15883249 TI - Successful treatment of relapsed acute promyelocytic leukemia in a patient receiving continuous ambulatory peritoneal dialysis with oral arsenic trioxide. PMID- 15883250 TI - Asians need different criteria for defining overweight and obesity. PMID- 15883251 TI - All calories are not equal. PMID- 15883252 TI - Serum ferritin levels in patients with nonalcoholic fatty liver disease. PMID- 15883253 TI - Is a low-carb, low-fat diet optimal? PMID- 15883254 TI - Confounders of uric acid levels. PMID- 15883255 TI - The uncertain natural history of thyrotoxic patients treated with combination interferon alfa-2beta and ribavirin. PMID- 15883256 TI - Leber hereditary optic neuropathy: biochemical lights in a blurry scenario. PMID- 15883257 TI - Internuclear ophthalmoplegia: unusual causes in 114 of 410 patients. AB - Internuclear ophthalmoplegia (INO) is a sign of exquisite localizing value, often due to either multiple sclerosis or infarction. To demonstrate that unusual causes of INO are more common than the 11% reported in previous series, this review considers a case series of 410 inpatients whom I personally examined during a 33-year period. In this series, the cause of INO was infarction in 157 patients (38%), multiple sclerosis in 139 (34%), and unusual causes in 114 (28%). Unusual causes included trauma (20 cases), tentorial herniation (20 cases), infection (17 cases), tumor (17 cases), iatrogenic injury (12 cases), hemorrhage (13 cases), vasculitis (7 cases), and miscellaneous (8 cases). Internuclear ophthalmoplegia was unilateral in 136 of the infarct cases (87%), 38 of those with multiple sclerosis (27%), and 48 of the unusual cases (42%). Because unusual causes compose more than one quarter of the cases, the differential diagnosis of INO should be tripartite: multiple sclerosis, stroke, and other causes. PMID- 15883258 TI - Metabolic patterns associated with the clinical response to galantamine therapy: a fludeoxyglucose f 18 positron emission tomographic study. AB - BACKGROUND: Regional brain correlates of treatment with cholinesterase inhibitors in those with Alzheimer disease are unknown. OBJECTIVE: To map brain metabolism associated with the treatment response to galantamine with fludeoxyglucose F 18 positron emission tomography in patients with Alzheimer disease. DESIGN: This is a hypothesis-driven, prospective, open-label study of 19 patients with mild to moderate Alzheimer disease examined before and after treatment with the cholinesterase inhibitor galantamine. Clinical examinations included the cognitive portion of the Alzheimer Disease Assessment Scale, the Mini-Mental State Examination, and the Neuropsychiatric Inventory. Imaging was performed using fludeoxyglucose F 18 positron emission tomography. The positron emission tomographic data, registered to a probabilistic anatomical atlas, were subjected to a voxel-based analysis of 3 subgroups: total patient analysis, cognitive analysis, and behavioral analysis. Subvolume thresholding corrected random lobar noise to produce 3-dimensional significance maps. RESULTS: The total group analysis showed an increase in left caudate metabolism with no significant change in clinical outcomes for the total group with treatment. Subgroup analysis of cognitive and behavioral responders demonstrated a significant activation of a striatal-thalamofrontal network with galantamine treatment that was not present in patients whose condition worsened or was unchanged by therapy. In cognitive subgroups, change in left anterior cingulate metabolism significantly correlated with change in the cognitive portion of the Alzheimer Disease Assessment Scale (r = 0.70, P = .02); in behavioral subgroups, right cingulate metabolic change significantly correlated with improvement in depression and right ventral putamen metabolic change with improvement in apathy (r = 0.63, P<.05 for both). CONCLUSION: Cognitive and behavioral responders to galantamine therapy show clinically related improvements in prefrontal network metabolism along with thalamic activation. PMID- 15883259 TI - Severe impairment of complex I-driven adenosine triphosphate synthesis in leber hereditary optic neuropathy cybrids. AB - BACKGROUND: Leber hereditary optic neuropathy (LHON) is a maternally inherited form of central vision loss associated with mitochondrial DNA point mutations that affect the ND subunits of complex I. OBJECTIVE: To elucidate the bioenergetic consequences of complex I dysfunction in LHON. DESIGN: The biochemical phenotypes of LHON mutations have been investigated using the transmitochondrial cytoplasmic hybrid (cybrid) cell model derived from the osteocarcoma parental cell line 143B.TK-. SETTING: Research laboratories at neuroscience and biochemistry departments at the University of Bologna, Scientific Institute "E. Medea," and University of College Medical School. PARTICIPANTS: Fibroblast cell lines were obtained from patients affected with LHON, as defined by the presence of 1 pathogenic mutation, and from healthy volunteers as controls to construct cybrid cell lines. MAIN OUTCOME MEASURES: Complex I (glutamate-malate)- and complex II (succinate)-dependent adenosine triphosphate (ATP) synthesis, their respective respiratory rates, and total cellular ATP content were investigated using digitonin permeabilized cybrid cells. Multiple cybrid cell lines were constructed, introducing into osteosarcoma derived rho(0) cells either wild-type or LHON mutant mitochondria carrying each of the 3 common mutations at positions 11778/ND4, 3460/ND1, and 14484/ND6. RESULTS: All 3 LHON mutations impaired ATP synthesis and the respiratory control ratio driven by complex I substrates. In contrast, succinate-driven ATP synthesis, respiration rates, and respiratory control ratios were not affected. However, the defective ATP synthesis with complex I substrates did not result in reduced ATP cellular content, indicating a compensatory mechanism. CONCLUSIONS: The LHON pathogenic mutations profoundly impair complex I-dependent synthesis of ATP, providing a common biochemical feature that may play a major role in LHON pathogenesis. Stratification of the results by mutation suggests that the 11778/ND4 mutation may induce an uncoupling of cybrid respiration, whereas the other 2 mutations impair the oxygen consumption rate. PMID- 15883260 TI - Brain magnetic resonance imaging findings in patients with mitochondrial cytopathies. AB - BACKGROUND: Mitochondrial cytopathies (MCs) are a heterogeneous group of clinical entities, some of which have classic phenotypes. Magnetic resonance imaging (MRI) has been reported to be helpful in the diagnosis of MC. OBJECTIVE: To correlate the most common brain MRI findings reported in patients with MC with the clinical findings in patients in different MC subgroups. DESIGN: Case series. SETTING: Patients with MCs seen at the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico. PATIENTS: Twenty-one patients with MC with the following phenotypes: chronic progressive external ophthalmoplegia (n = 7), Kearns-Sayre syndrome (n = 7), mitochondrial neurogastrointestinal encephalopathy (n = 6), and myoclonic epilepsy with ragged red fiber myopathy (n = 1). RESULTS: Brain MRI abnormalities were found in 20 (95%) of 21 patients. The most frequent abnormalities were widespread white matter hyperintensity in 19 patients (90%), supratentorial cortical atrophy in 18 patients (86%), and cerebellar atrophy in 13 patients (62%). Widespread white matter hyperintensity (P<.001) and supratentorial cortical atrophy (P = .001) were each correlated significantly with MC. Subsequent subgroup analyses showed that the absence of basal ganglia hyperintensity was correlated with Kearns-Sayre syndrome (P < .001) and the presence of supratentorial cortical atrophy was correlated with mitochondrial neurogastrointestinal encephalopathy (P = .005). CONCLUSIONS: The presence of widespread white matter hyperintensity and/or supratentorial cortical atrophy in brain MRI may help to establish the diagnosis of MC. The radiologist has a role to play in the workup of MC by confirming the diagnosis and possibly distinguishing different subgroups of MC. PMID- 15883261 TI - New DGK gene mutations in the hepatocerebral form of mitochondrial DNA depletion syndrome. AB - OBJECTIVE: To document novel homozygous mutations in the gene for deoxyguanosine kinase (DGK) in 3 children with mitochondrial DNA depletion. DESIGN: Clinical features included liver failure, hypotonia, and nystagmus in 2 siblings, and liver cirrhosis, optic dysplasia, nystagmus, and microcephaly in the third patient. We sequenced the whole coding region of the DGK gene. RESULTS: We identified 2 novel homozygous mutations, G352A and C269T, that lead to truncated proteins. CONCLUSION: These data confirm that DGK mutations typically affect the liver and brain. PMID- 15883262 TI - Atorvastatin for the treatment of mild to moderate Alzheimer disease: preliminary results. AB - BACKGROUND: Laboratory evidence of cholesterol-induced production of amyloid beta as a putative neurotoxin precipitating Alzheimer disease, along with epidemiological evidence, suggests that cholesterol-lowering statin drugs may favorably influence the progression of the disorder. OBJECTIVE: To determine if treatment with atorvastatin calcium affects the cognitive and/or behavioral decline in patients with mild to moderate Alzheimer disease. DESIGN: Pilot intention-to-treat, proof-of-concept, double-blind, placebo-controlled, randomized (1:1) trial with a 1-year exposure to once-daily atorvastatin calcium (80 mg; two 40-mg tablets) or placebo using last observation carried forward analysis of covariance as the primary method of statistical assessment. PARTICIPANTS: Individuals with mild to moderate Alzheimer disease (Mini-Mental State Examination score of 12-28) were recruited. Of the 98 participants providing informed consent, 71 were eligible for randomization, 67 were randomized, and 63 subjects completed the 3-month visit and were considered evaluable. MAIN OUTCOME MEASURES: The primary outcome measures were change in Alzheimer's Disease Assessment Scale-cognitive subscale and the Clinical Global Impression of Change Scale scores. The secondary outcome measures included scores on the Mini-Mental State Examination, Geriatric Depression Scale, the Neuropsychiatric Inventory Scale, and the Alzheimer's Disease Cooperative Study Activities of Daily Living Inventory. The tertiary outcome measures included total cholesterol, low-density lipoprotein cholesterol, and very low-density lipoprotein cholesterol levels. RESULTS: Atorvastatin reduced circulating cholesterol levels and produced a positive signal on each of the clinical outcome measures compared with placebo. This beneficial effect reached significance for the Geriatric Depression Scale and the Alzheimer's Disease Assessment Scale cognitive subscale at 6 months and was significant at the level of a trend for the Alzheimer's Disease Assessment Scale-cognitive subscale, Clinical Global Impression of Change Scale, and Neuropsychiatric Inventory Scale at 12 months assessed by analysis of covariance with last observation carried forward. CONCLUSION: Atorvastatin treatment may be of some clinical benefit and could be established as an effective therapy for Alzheimer disease if the current findings are substantiated by a much larger multicenter trial. PMID- 15883263 TI - Predictors of preclinical Alzheimer disease and dementia: a clinicopathologic study. AB - BACKGROUND: To understand the earliest signs of cognitive decline caused by Alzheimer disease (AD) and other illnesses causing dementia, information is needed from well-characterized individuals without dementia studied longitudinally until autopsy. OBJECTIVE: To determine clinical and cognitive features associated with the development of AD or other dementias in older adults. DESIGN: Longitudinal study of memory and aging. SETTING: Alzheimer's Disease Research Center, St Louis, Mo. MAIN OUTCOME MEASURES: Clinical Dementia Rating, its sum of boxes, and neuropathologic diagnosis of dementia. PARTICIPANTS: Eighty control participants who eventually came to autopsy. RESULTS: Individuals who did not develop dementia showed stable cognitive performance. Entry predictors of dementia were age, deficits in problem solving as well as memory, slowed psychomotor performance, and depressive features. Minimal cognitive decline occurred prior to dementia diagnosis, after which sharp decline was noted. Even individuals who were minimally cognitively impaired (Clinical Dementia Rating = 0.5) typically had neuropathologic AD at autopsy. Histopathologic AD also was present in 34% of individuals who did not have dementia at death; these individuals without dementia showed an absence of practice effects on cognitive testing. CONCLUSIONS: Increased age, depressive features, and even minimal cognitive impairment, as determined clinically by Clinical Dementia Rating sum of boxes and by slowed psychomotor performance, identify older individuals without dementia who develop dementia. Older adults who do not develop dementia have stable cognitive performance. The absence of practice effects may denote the subset of older adults without dementia with histopathologic AD, which may reflect a preclinical stage of the illness. PMID- 15883264 TI - Correlation of cerebrospinal fluid levels of tau protein phosphorylated at threonine 231 with rates of hippocampal atrophy in Alzheimer disease. AB - BACKGROUND: The microtubule-associated tau protein abnormally phosphorylated at threonine 231 (p-tau231) has been investigated as a potential marker of Alzheimer disease. Levels of cerebrospinal fluid (CSF) p-tau231 vary across patients with Alzheimer disease. We hypothesized that these variations partially reflect differences in the degree of neuronal damage and therefore may be used to predict structural disease progression. OBJECTIVE: To investigate whether CSF p-tau231 levels correlate with rates of hippocampal atrophy as an in vivo marker of regional neuronal loss. DESIGN AND PATIENTS: We measured hippocampal volumes on the basis of serial magnetic resonance image examinations in 22 patients with Alzheimer disease. In addition, we determined CSF p-tau231 levels at baseline. RESULTS: Levels of CSF p-tau231 were significantly correlated with baseline hippocampal volumes (P<.001) and rates of hippocampal atrophy (left hippocampus, P<.001; right hippocampus, P = .02), independent of disease duration and severity. CONCLUSION: These findings suggest that variations in p-tau231 levels may be used to predict progression of brain atrophy in patients with Alzheimer disease. PMID- 15883265 TI - Earlier onset of Alzheimer disease symptoms in latino individuals compared with anglo individuals. AB - BACKGROUND: Latino individuals are the largest minority group and the fastest growing population group in the United States, yet there are few studies comparing the clinical features of Alzheimer disease (AD) in this population with those found in Anglo (white non-Latino) patients. OBJECTIVE: To compare the age at AD symptom onset in Latino and Anglo individuals. DESIGN: Cross-sectional assessment using standardized methods to collect and compare age at AD symptom onset, demographic variables, and medical variables. SETTING: Five National Institute on Aging-sponsored Alzheimer's Disease Centers with experience evaluating Spanish-speaking individuals. PATIENTS: We evaluated 119 Latino and 55 Anglo patients who had a diagnosis of AD. MAIN OUTCOME MEASURE: Age at symptom onset. RESULTS: After adjusting for center, sex, and years of education, Latino patients had a mean age at symptom onset 6.8 years earlier (95% confidence interval, 3.5-10.3 years earlier) than Anglo patients. CONCLUSIONS: An earlier age at symptom onset suggests that US mainland Latino individuals may experience an increased burden of AD compared with Anglo individuals. The basis for the younger age at symptom onset remains obscure. PMID- 15883266 TI - Alzheimer disease and mortality: a 15-year epidemiological study. AB - BACKGROUND: Alzheimer disease (AD) is considered a leading cause of death, but few studies have examined the contribution of AD to mortality based on follow-up of representative US cohorts. OBJECTIVE: To examine mortality rates, duration of survival, causes of death, and the contribution of AD to the risk of mortality in an aging community-based cohort, controlling for other predictors. DESIGN: Fifteen-year prospective epidemiological study. Mortality rates per 1000 person years and the population-attributable risk of mortality were determined. Cox proportional hazards models were used to estimate relative risk of mortality due to AD, adjusting for relevant covariates. Death certificates were abstracted for listed causes of death. SETTING: A largely blue-collar rural community in southwestern Pennsylvania. PARTICIPANTS: A community-based cohort of 1670 adults 65 years and older at study enrollment. MAIN OUTCOME MEASURE: Mortality. RESULTS: In the overall cohort, AD was a significant predictor of mortality, with a hazard ratio of 1.4 after adjusting for covariates. The population-attributable risk of mortality from AD was 4.9% based on the same model. Examining the sexes separately, AD increased mortality risk only among women. Death certificates of AD subjects were more likely to list dementia/AD, other brain disorders, pneumonia, and dehydration, and less likely to include cancer. CONCLUSIONS: Alzheimer disease was responsible for 4.9% of the deaths in this elderly cohort. Alzheimer disease increased the risk of mortality 40% in the cohort as a whole and separately in women but not in men. The mean (SD) duration of survival with AD was 5.9 (3.7) years, and longer with earlier age at onset. PMID- 15883267 TI - Enhanced benefit of increasing interferon beta-1a dose and frequency in relapsing multiple sclerosis: the EVIDENCE Study. AB - BACKGROUND: The EVIDENCE (Evidence of Interferon Dose-Response: European North American Comparative Efficacy) Study demonstrated that patients with multiple sclerosis (MS) who initiate interferon beta-1a therapy with 44 microg 3 times weekly (TIW) were less likely to have a relapse or activity on magnetic resonance imaging (MRI) compared with those who initiate therapy at a dosage of 30 microg 1 time weekly (QW). OBJECTIVE: To determine the effect of changing the dosage from 30 microg QW to 44 microg TIW in this extension of the EVIDENCE Study. DESIGN/PATIENTS: Patients with relapsing MS originally randomized to interferon beta-1a, 30 microg QW, during the comparative phase of the study changed to 44 microg TIW, whereas patients originally randomized to 44 microg TIW continued that regimen. Patients were followed up, on average, for an additional 32 weeks. MAIN OUTCOME MEASURE: The within-patient pretransition to post-transition change in relapse rate. RESULTS: At the transition visit, 223 (73%) of 306 patients receiving 30 microg QW converted to 44 microg TIW, and 272 (91%) of 299 receiving 44-microg TIW continued the same therapy. The post-transition annualized relapse rate decreased from 0.64 to 0.32 for patients increasing the dose (P<.001) and from 0.46 to 0.34 for patients continuing 44-microg TIW (P = .03). The change was greater in those increasing dose and frequency (P = .047). Patients converting to the 44-mug TIW regimen had fewer active lesions on T2-weighted MRI compared with before the transition (P = .02), whereas those continuing the 44-microg TIW regimen had no significant change in T2 active lesions. Patients who converted to high-dose/high-frequency interferon beta-1a therapy had increased rates of adverse events and treatment terminations consistent with the initiation of high dose subcutaneous interferon therapy. CONCLUSIONS: Patients receiving interferon beta-1a improved on clinical and MRI disease measures when they changed from 30 microg QW to 44 microg TIW. PMID- 15883268 TI - Kinin B1 receptor expression on multiple sclerosis mononuclear cells: correlation with magnetic resonance imaging T2-weighted lesion volume and clinical disability. AB - BACKGROUND: We have previously shown that the inducible kinin B(1) receptor is expressed on T lymphocytes during relapses and progression in multiple sclerosis. OBJECTIVE: To evaluate the correlation between the expression of B1 receptor on peripheral blood mononuclear cells derived from patients who have multiple sclerosis with serial, clinical magnetic resonance imaging and immunological study-derived measures. DESIGN: Using frozen samples obtained from a high frequency magnetic resonance imaging-immunological study, we analyzed B1 receptor messenger RNA (mRNA) expression in peripheral blood-derived mononuclear cells serially collected from 6 patients with multiple sclerosis and 8 healthy control subjects by semiquantitative radioactive duplex reverse transcriptase-polymerase chain reaction amplification. Time-course kinin B1-actin mRNA ratios were subsequently compared with corresponding clinical magnetic resonance imaging and immune parameters. RESULTS: The time-course kinin B1-actin mRNA ratio correlated positively with the Expanded Disability Status Scale index (P<.001), occurrence of clinical relapse (P = .02), volume of lesion on T2-weighted images (P<.003) and interleukin 2 receptor and major histocompatibility complex class II expression on CD4+ lymphocytes, but not with gadolinium-enhancing lesions. The time-course kinin B1-actin mRNA ratios were 5 to 25 times lower in samples derived from healthy controls. CONCLUSION: The correlation of kinin B1 receptor mRNA levels with dynamic clinical and magnetic resonance imaging measures suggests that expression of this receptor can serve as an index of disease activity in multiple sclerosis. PMID- 15883269 TI - Diffusion-tensor magnetic resonance imaging detects normal-appearing white matter damage unrelated to short-term disease activity in patients at the earliest clinical stage of multiple sclerosis. AB - BACKGROUND: Diffusion-tensor (DT) magnetic resonance imaging (MRI) has the potential to elucidate some characteristics of tissue microstructure inaccessible to other MRI techniques. OBJECTIVE: To investigate whether normal-appearing brain tissue abnormalities occur in patients with multiple sclerosis at the earliest clinical stage and whether their severity is predictive of a short-term disease evolution by using DT MRI. DESIGN: Forty-five patients and 22 healthy control subjects were studied. All patients had had a clinically isolated syndrome within the 3 months preceding study enrollment and paraclinical evidence of disease dissemination in space. During a single session, dual-echo, pulsed-gradient spin echo echo-planar, and postgadolinium T1-weighted images of the brain were obtained from each subject. In patients, dual-echo and enhanced images were obtained after 3 and 12 months, to detect MRI signs of disease dissemination in time. An on-study neurological examination was also conducted to ascertain the occurrence of clinical relapses. Mean diffusivity and fractional anisotropy maps were derived from DT images. Normal-appearing white matter (NAWM) and normal appearing gray matter mean diffusivity and fractional anisotropy histograms were produced and analyzed. RESULTS: During the study period, 29 patients showed MRI evidence of disease dissemination in time. When compared with healthy controls, patients showed higher average NAWM mean diffusivity (P = .01), lower average NAWM mean diffusivity peak height (P < .001), and fractional anisotropy (P < .001). The DT MRI characteristics of patients did not differ between those with and those without disease dissemination in time at follow-up. CONCLUSIONS: In patients with multiple sclerosis at the earliest clinical stage, the severity of NAWM damage does not predict new lesion formation in the short term, suggesting that the "diffuse" component of tissue damage is, at least partially, independent of the "discrete," predominantly inflammatory aspects of the disease since its clinical onset. PMID- 15883270 TI - Ipsilateral hemiparesis caused by a corona radiata infarct after a previous stroke on the opposite side. AB - Ipsilateral hemiparesis after a supratentorial stroke is rare. However, the role of the reorganization of the unaffected hemisphere in recovery after a stroke is poorly understood. Two patients developed ipsilateral hemiparesis after a left corona radiata infarct. Both of these patients had previously experienced contralateral hemiparesis after a right-sided supratentorial stroke. Functional magnetic resonance imaging demonstrated bilateral motor area activation during paretic left hand movement. This finding suggests that the ipsilateral hemiparesis was caused by a new stroke in the ipsilateral motor system that was functionally reorganized after the previous stroke. PMID- 15883271 TI - Brachial amyotrophic diplegia in a patient with human immunodeficiency virus infection: widening the spectrum of motor neuron diseases occurring with the human immunodeficiency virus. AB - Although amyotrophic lateral sclerosis and progressive spinal muscular atrophy have been recognized to occur in association with human immunodeficiency virus infection, to our knowledge, brachial amyotrophic diplegia, a form of segmental motor neuron disease, has not been previously reported. Brachial amyotrophic diplegia results in severe lower motor neuron weakness and atrophy of the upper extremities in the absence of bulbar or lower extremity involvement, pyramidal features, bowel and bladder incontinence, and sensory loss. We describe a human immunodeficiency virus-seropositive man without severe immunosuppression or prior AIDS-defining illnesses who had brachial amyotrophic diplegia. This disorder may represent one end of a spectrum of motor neuron diseases occurring with this retrovirus infection. PMID- 15883272 TI - Otitic hydrocephalus revisited. PMID- 15883273 TI - Compression of the deep motor branch of the ulnar nerve: an unusual cause of pure motor neuropathy and hand wasting. PMID- 15883274 TI - Cogwheel rigidity. PMID- 15883275 TI - Commentary regarding neoplastic meningitis secondary to multiple myeloma. PMID- 15883276 TI - Etiology of mirror writing in Japanese. PMID- 15883277 TI - Second primary cancers after enrollment in the COMS trials for treatment of choroidal melanoma: COMS Report No. 25. AB - OBJECTIVE: To report sites of second primary cancer and the time to first diagnosis during routine follow-up after treatment for choroidal melanoma. DESIGN: Prospective longitudinal follow-up of patients enrolled in 2 randomized trials conducted by the Collaborative Ocular Melanoma Study (COMS) Group. METHODS: Baseline and annual or semiannual systemic and laboratory evaluations were performed according to a standard protocol for 2320 patients enrolled in the COMS without evidence of melanoma metastasis or other primary cancer at baseline. Deaths were coded by a mortality coding committee. RESULTS: Subsequent to treatment for choroidal melanoma, a total of 222 patients were diagnosed with a second primary cancer other than basal or squamous cell skin cancer (5-year rate of 7.7% [95% confidence interval, 6.6%-9.0%]). The most common sites were prostate (23% of reported cases) and breast (17%); 12 of these 222 patients were diagnosed simultaneously with second primary cancers in 2 or more sites. Of these 222 patients, 113 died; 37 (33%) were coded as dead with melanoma metastasis, 33 (29%) as dead with a malignant tumor other than metastatic melanoma, and 13 (11%) as dead with a malignancy of uncertain origin. Radiotherapy did not significantly increase the development of second primary cancers. The rate of diagnosis of second primary cancer did not differ significantly by smoking status, although the rate in former smokers was increased vs that observed in either current smokers or those who never smoked. CONCLUSION: Routine medical surveillance for development of second primary cancers among patients treated for choroidal melanoma is important, especially for those with a history of smoking, regardless of the size of choroidal melanoma at the time of treatment. PMID- 15883278 TI - Incidence of acute endophthalmitis following penetrating keratoplasty: a systematic review. AB - OBJECTIVE: To determine the incidence of acute endophthalmitis following penetrating keratoplasty (PK) over time. METHODS: A systematic review of English language articles was conducted by performing a broad search of the PubMed database from 1963 through March 2003 using such keywords as penetrating keratoplasty, endophthalmitis, and postoperative complication. Additional studies were identified from bibliographies of relevant articles and published proceedings. The proportion of eyes with acute endophthalmitis as a postoperative complication was recorded, and pooled incidence rates were assessed over time. RESULTS: From 1870 unique, potentially relevant citations, 66 original studies that addressed endophthalmitis and met the selection criteria were analyzed. A total of 90 549 PKs were pooled, resulting in an overall estimate of 0.382% post PK endophthalmitis, but a change over time was noted. The rate of endophthalmitis was 0.200% in the 2000-2003 period, 0.453% in the 1990s, 0.376% in the 1980s, and 0.142% during the 1970s. Furthermore, a downward trend in the incidence of endophthalmitis after 1992 was observed compared with 1991 and earlier. CONCLUSIONS: This systematic review indicates that the incidence of endophthalmitis associated with PK has declined during the last decade. PMID- 15883279 TI - Acute endophthalmitis following cataract surgery: a systematic review of the literature. AB - OBJECTIVES: To determine the reported incidence of acute endophthalmitis following cataract extraction over time and to explore possible contributing factors, such as type of cataract incision. METHODS: A systematic review of English-language articles was conducted by performing a broad search of PubMed from 1963 through March 2003 using such terms as cataract extraction, endophthalmitis, and postoperative complication. Additional studies were identified from bibliographies of relevant articles and published proceedings. Surgical approach was recorded, when available. Pooled incidence rates and relative risks of developing endophthalmitis using different incision techniques were assessed. RESULTS: From 4916 unique, potentially relevant citations, 215 studies that addressed endophthalmitis and met the selection criteria were analyzed. A total of 3 140 650 cataract extractions were pooled resulting in an overall rate of 0.128% of postcataract endophthalmitis. However, the incidence of acute endophthalmitis changed over time, with a significant increase since 2000 compared with previous decades (relative risk, 2.44 [95% confidence interval, 2.27-2.61]). The rate of endophthalmitis was 0.265% in the 2000-2003 period, 0.087% in the 1990s, 0.158% in the 1980s, and 0.327% during the 1970s. Furthermore, an upward trend in rates after 1992 was noted, compared with 1991 and prior. Incision type appeared to significantly influence risk, as endophthalmitis following clear corneal cataract extraction during the 1992-2003 period was 0.189% compared with 0.074% (relative risk, 2.55 [95% confidence interval, 1.75-3.71]) for scleral incision and 0.062% (relative risk, 3.06 [95% confidence interval, 2.48-3.76]) for limbal incision. CONCLUSIONS: This systematic review indicates that the incidence of endophthalmitis associated with cataract extraction has increased over the last decade. This upward trend in endophthalmitis frequency coincides temporally with the development of sutureless clear corneal incisions. PMID- 15883280 TI - Near confluent laser photocoagulation for the treatment of threshold retinopathy of prematurity. AB - OBJECTIVE: To evaluate the effect of a near confluent pattern of indirect laser photocoagulation in reducing the rate of progression and re-treatment of threshold retinopathy of prematurity. METHODS: This study examined a noncomparative interventional case series. We performed a retrospective review of the medical records of patients who underwent peripheral laser ablation by 1 surgeon for threshold retinopathy of prematurity from 1997 to 2002. A total of 58 eyes from 31 patients were treated, and 44 eyes of 23 patients were included in the study. Ten eyes of 5 infants had zone 1 disease, and 34 eyes of 18 infants had zone 2 disease. Laser spots were placed in a near confluent pattern in the peripheral avascular retina between the ridge of extraretinal proliferation and the ora serrata. The mean +/- SD number of laser spots was 2534 +/- 455 for zone 1 (range, 2100-3378) and 1850 +/- 487 for zone 2 (range, 1030-2689). RESULTS: In 7 eyes of 4 infants with zone 1 disease, the retinopathy regressed and did not require any further treatment. Three eyes of 2 infants, however, progressed after laser treatment and required vitrectomy surgery. Progression was defined as the development of stage 4 or 5 disease. None of the patients with zone 2 disease had progression of retinopathy, and none of them needed more than 1 treatment. Patients tolerated the procedure well, and there were no complications at the time of the procedure or at follow-up visits. CONCLUSIONS: A near confluent pattern of laser photocoagulation may reduce the rate of progression of threshold retinopathy of prematurity in zone 2 (0%). The near confluent pattern of treatment may also reduce the re-treatment rate of the disease (0%). Larger studies are needed to confirm our findings. PMID- 15883281 TI - Retinopathy from a green laser pointer: a clinicopathologic study. AB - OBJECTIVE: To report retinopathy following exposure to light from a commercially available class 3A green laser pointer. METHODS: A 55-year-old woman with a ring melanoma was scheduled for enucleation. The eye (visual acuity 20/20) had a healthy-appearing macular retina. The retina was exposed to light from a commercially available class 3A green laser: 60 seconds to the fovea, 5 minutes to a site 5 degrees below the fovea, and 15 minutes to a site 5 degrees superior to the fovea. Color photographs were obtained before and after exposure. The eye was enucleated 20 days after exposure. RESULTS: Laser power measurements averaged less than 5 mW. Retinopathy was observed 24 hours after laser exposure. This was characterized by a yellowish discoloration at the level of the retinal pigment epithelium (RPE) in the subfoveal region and at the site superior to the macula where the retina received 15 minutes of laser exposure. Each site developed granular changes at the level of the RPE within 5 days of exposure. Histologic study showed RPE damage in the exposed subfoveal and parafoveal regions. CONCLUSION: A class 3A green laser pointer caused visible retinopathy in the human eye with exposures as short as 60 seconds. PMID- 15883282 TI - Cyclosporine vs tacrolimus therapy for posterior and intermediate uveitis. AB - OBJECTIVES: To compare the efficacy and tolerability of tacrolimus and cyclosporine therapy for noninfectious posterior segment intraocular inflammation and to evaluate their effect on peripheral blood CD4(+) T-cell phenotype and activation status. METHODS: Thirty-seven patients who required second-line immunosuppression for posterior segment intraocular inflammation were enrolled in this prospective randomized trial of tacrolimus vs cyclosporine therapy. The main outcome measures were visual acuity, binocular indirect ophthalmoscopy score, adverse effects, and quality of life. In addition, peripheral blood CD4(+) T-cell phenotype and activation status were evaluated by flow cytometry before treatment and at 2, 4, and 12 weeks using CD69, chemokine receptor (CCR4, CCR5, and CXCR3), and intracellular cytokine (tumor necrosis factor alpha, interferon-gamma, and interleukin 10) expression. RESULTS: Thirteen patients (68%) taking tacrolimus and 12 patients (67%) taking cyclosporine responded to treatment. Cyclosporine therapy was associated with a higher incidence of reported adverse effects. Mean arterial pressure and serum cholesterol level were significantly higher at 3 months in the cyclosporine group than the tacrolimus group. No significant difference was detected with regard to effect on quality of life or CD4(+) T-cell phenotype. CONCLUSIONS: Tacrolimus and cyclosporine were similar with regard to efficacy for posterior segment intraocular inflammation, but the results suggested a more favorable safety profile for tacrolimus therapy. PMID- 15883283 TI - Ingress of India ink into the anterior chamber through sutureless clear corneal cataract wounds. AB - BACKGROUND: Sutureless clear corneal cataract incisions may be associated with an increased risk of endophthalmitis. OBJECTIVE: To assess the degree of ocular surface fluid ingress into the anterior chamber of cadaveric human globes with clear corneal wounds. METHODS: Self-sealing clear corneal incisions were created in 4 eyes, and intraocular pressure was controlled with an infusion cannula. To evaluate possible flow of surface fluid through the corneal wound, india ink was applied to the corneal surface while the intraocular pressure was varied, so as to simulate the intraocular pressure fluctuations secondary to blinking or eye squeezing. The optical density from aqueous samples of globes were measured both before and after india ink application using a spectrophotometer. RESULTS: Aqueous aspirates from the 3 globes with sutureless clear corneal wounds revealed a significant increase in spectrophotometric readings (P<.01), in contrast to the sutured wound, which did not show an increase in absorbance level relative to the baseline. Ink particles were both grossly and microscopically visible inside the sutureless corneal wounds. CONCLUSIONS: Fluctuations of intraocular pressure following sutureless clear corneal cataract surgery may allow entry of surface fluid into the anterior chamber during the initial postoperative period when the wound is not healed. PMID- 15883284 TI - Effect of intravitreal triamcinolone acetonide on susceptibility to experimental bacterial endophthalmitis and subsequent response to treatment. AB - OBJECTIVES: Recent reports of high endophthalmitis rates after intravitreal triamcinolone acetonide injection have raised concerns about the safety of this treatment. We sought to evaluate the effect of intravitreal triamcinolone injection on (1) the susceptibility to experimental bacterial endophthalmitis and (2) the subsequent therapeutic response to antibiotic treatment. DESIGN: For the susceptibility study, the right eye of 40 New Zealand white rabbits received an intravitreal injection of a known quantity of Staphylococcus epidermidis organisms. Half of the eyes received a simultaneous intravitreal injection of triamcinolone acetonide, 4 mg. All eyes were examined daily for signs of endophthalmitis (photophobia, conjunctival injection, and vitritis) using standardized grading protocols (scaled from 0 to 4 with increasing severity). On day 7, vitreous cultures were obtained. For the therapeutic response study, the right eye of 12 rabbits received an intravitreal injection of S epidermidis organisms sensitive to vancomycin. Half of the eyes received a simultaneous intravitreal injection of triamcinolone acetonide, 4 mg. All 12 eyes received an intravitreal injection of vancomycin hydrochloride, 1 mg, on development of the first signs of endophthalmitis. All eyes were examined daily for 7 additional days. On day 7 after treatment, vitreous cultures were obtained. RESULTS: In the susceptibility study, all 40 eyes developed signs of endophthalmitis. In eyes that received intravitreal bacteria plus triamcinolone, 17 (85%) of the 20 vitreous cultures were positive, whereas only 6 (30%) were positive in the 20 eyes receiving bacteria alone (P = .001). The vitritis was significantly increased in the bacteria plus triamcinolone group compared with the bacteria only group (17 of 20 vs 7 of 20 with 4+ vitritis, respectively; P = .003). In the therapeutic response study, all 12 eyes developed clinical signs of endophthalmitis within 48 hours. All vitreous samples obtained 7 days after intravitreal vancomycin injection were culture negative. However, the severity of vitritis at the time of vitreous sampling was less in the eyes receiving triamcinolone plus bacteria compared with eyes receiving bacteria alone (0 of 6 vs 5 of 6 with 4+ vitritis, respectively; P = .02). CONCLUSIONS: In eyes with experimentally induced bacterial endophthalmitis, the presence of intravitreal triamcinolone results in a higher culture-positive rate and a higher degree of inflammation, suggesting an impaired ocular immune response and greater susceptibility to infection. However, in eyes with experimentally induced bacterial endophthalmitis receiving early treatment with intravitreal antibiotics, triamcinolone appears to suppress the ocular inflammatory response without impairing the therapeutic effect. CLINICAL RELEVANCE: These data suggest that caution must be exercised when combining intravitreal triamcinolone injection with intraocular surgery. PMID- 15883285 TI - Resistance and susceptibility of human uveal melanoma cells to TRAIL-induced apoptosis. AB - OBJECTIVE: To evaluate the resistance and susceptibility of human uveal melanoma cells to apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). METHODS: The sensitivity of 11 human uveal melanoma cell lines was analyzed by flow cytometry for the expression of TRAIL receptors and the antiapoptotic protein survivin. Caspase-8 and caspase-10 expression was also examined using reverse transcriptase-polymerase chain reaction and Western blot analysis. RESULTS: Only 4 melanoma cell lines were sensitive to TRAIL-induced apoptosis. Positive correlation was observed between resistance and expression of survivin. Up-regulation of survivin by gene transfer enhanced resistance to TRAIL induced apoptosis, whereas transfection with survivin antisense rendered resistant melanoma cells susceptible to TRAIL-induced apoptosis. Survivin expression and susceptibility to TRAIL-induced apoptosis could also be manipulated by treatment with actinomycin D, which produced a 30% to 50% decrease in the expression of survivin (P < .01) and a 5- to-7-fold increase in TRAIL induced apoptosis (P < .001). CONCLUSIONS: Resistance of uveal melanoma cells to TRAIL-induced apoptosis is regulated by antiapoptotic proteins such as survivin. CLINICAL RELEVANCE: Manipulation of apoptosis regulatory proteins, such as survivin, may have therapeutic applications in the management of uveal melanomas. PMID- 15883286 TI - Incidence of emergency department-treated eye injury in the United States. AB - OBJECTIVE: To present the descriptive epidemiology of emergency department treated eye injury in the United States. METHODS: Data from the Consumer Product Safety Commission's National Electronic Injury Surveillance System All Injury Program was used to estimate the number of patients treated in US emergency departments for eye injuries. Eye injury rates were calculated according to age, sex, and race, and characteristics of the injury event were determined. RESULTS: The rate of emergency department-treated eye injury in the United States is 3.15 per 1000 population (95% confidence interval, 2.66-3.63). Rates were highest among those in their 20s and 30s, among males, and among American Indians and African Americans. The majority of injuries occurred in the home, and contusions and abrasions were the most common types of injury. CONCLUSION: This study documents the burden of eye injury in the United States and identifies areas wherein future research activities should be directed. PMID- 15883287 TI - Need for eye care among older adults with diabetes mellitus in fee-for-service and managed Medicare. AB - OBJECTIVE: To compare rates of need for eye care among Medicare beneficiaries with network-model Medicare+Choice (MC) and fee-for-service (FFS) health insurance. METHODS: Cross-sectional study of a random sample of MC and FFS community-dwelling Medicare beneficiaries with diabetes who are older than 65 years of age in Los Angeles County. Study ophthalmologists masked to the participants' type of health insurance performed standardized dilated eye examinations and indicated the need for ophthalmic care during the next 6 months. To evaluate the association between type of insurance and need for treatment, we constructed logistic regression models adjusted for participant sociodemographic and clinical characteristics. RESULTS: The 311 MC and 107 FFS respondents reported comparable rates of eye care provider visits and preexisting eye diseases. However, on masked clinical examination, MC respondents were more likely to have diabetic retinopathy, visually significant cataract, glaucoma, or suspected glaucoma than FFS participants (68% vs 46%, P<.001). In multivariate analyses, persons enrolled in MC were significantly more likely than FFS participants to require further treatment during the next 6 months (42% vs 24%, P = .01). CONCLUSIONS: Data from standardized study ophthalmic examinations suggest high rates of unrecognized and untreated eye diseases among Medicare beneficiaries enrolled in both FFS and MC and significantly higher rates of need for care among MC participants. PMID- 15883288 TI - The Knapps. PMID- 15883289 TI - Noninfectious uveitis: a scarcity of randomized clinical trials. PMID- 15883290 TI - Meeting our ethical obligations in medical publishing: responsibilities of editors, authors, and readers of peer-reviewed journals. PMID- 15883291 TI - Systemic antibiotics for communitywide trachoma control. PMID- 15883292 TI - Orbital sclerosing extramedullary hematopoietic tumor. PMID- 15883293 TI - Limbal stem cell deficiency associated with LADD syndrome. PMID- 15883294 TI - Aggressive glioma of adulthood simulating ischemic optic neuropathy. PMID- 15883295 TI - Transient cortical blindness with leptomeningeal enhancement after attempted peripherally inserted central venous catheter placement. PMID- 15883296 TI - Disseminated Fusarium infection presenting as bilateral endogenous endophthalmitis in a patient with acute myeloid leukemia. PMID- 15883297 TI - Central retinal vein occlusion secondary to an intraocular nematode. PMID- 15883298 TI - Predictive value of slitlamp examinations in screening donor corneas for prior refractive surgery. PMID- 15883300 TI - Appropriate timing for informed-consent discussions. PMID- 15883302 TI - Viagra and anterior ischemic optic neuropathy. PMID- 15883303 TI - Retinal vessel caliber and diabetic retinopathy. PMID- 15883305 TI - Who can say yes (or no) to a physician--and how does the physician know they can? PMID- 15883306 TI - Is migraine a risk factor for stroke? PMID- 15883307 TI - Continued need for caution in the diagnosis of Duchenne muscular dystrophy. PMID- 15883308 TI - The genetic causes of basal ganglia calcification, dementia, and bone cysts: DAP12 and TREM2. AB - BACKGROUND: Polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy (PLOSL), or Nasu-Hakola disease, is a presenile dementia associated with loss of myelin, basal ganglia calcification, and bone cysts. It is caused by recessively inherited mutations in two genes encoding subunits of a cell membrane-associated receptor complex: TREM2 and DAP12. The clinical course of PLOSL has not been characterized in a series of patients with TREM2 mutations. METHODS: The authors compare neurologic and neuroradiologic follow-up data of six patients carrying TREM2 mutations with PLOSL due to defective DAP12 genes. The authors review the known mutations in these two genes. RESULTS: Mutations in DAP12 and TREM2 result in a uniform disease phenotype. In Finnish and Japanese patients with PLOSL, DAP12 mutations predominate, whereas TREM2 is mutated more frequently elsewhere. CONCLUSIONS: Polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy should be considered in adult patients under age 50 years with dementia and basal ganglia calcification. Radiographs of ankles and wrists, and DNA test in uncertain cases, confirm the diagnosis. PMID- 15883309 TI - Generalized arteriopathy in patients with cervical artery dissection. AB - OBJECTIVE: To make an ultrastructural comparison of superficial temporal artery (STA) biopsy specimens from patients with spontaneous cervical artery dissection (sCAD) and controls. METHODS: The authors used light microscopic examination of semithin sections and electron microscopic examination of ultrathin sections of STA biopsy specimens from patients with sCAD and controls. RESULTS: STA biopsy specimens from patients with sCAD taken around the time of the dissection showed a zone of connective tissue weakening with fissuring at the junction between the tunica media (TM) and the tunica adventitia (TA) in seven of nine specimens and erythrocyte infiltration in eight of nine specimens but in none of the control specimens. Light microscopy demonstrated transparent circular spots that, on electron microscopy, turned out to represent erythrocytes and other cellular components at different stages of degradation. Occasionally, scattered immune cells were found in specimens from patients with sCAD. In addition, smooth muscle cells of the synthetic phenotype, some of them showing extensive vacuolation were more common in the TM of STA biopsy specimens from patients with sCAD than in control specimens. CONCLUSIONS: Signs of tissue weakening along the TM/TA junction in STA biopsy specimens of patients with sCAD but not in controls suggest the presence of a generalized arteriopathy leading to impairment of the stability of the arterial wall in patients with sCAD. Limiting factors of the study are that some control biopsies were obtained from autopsies and that the anticoagulation status of patients and controls were not completely comparable. PMID- 15883310 TI - The ability of persons with Alzheimer disease (AD) to make a decision about taking an AD treatment. AB - OBJECTIVE: To examine the severity of impairments in the decision-making abilities (understanding, appreciation, reasoning, and choice) and competency to make a decision to use an Alzheimer disease (AD)-slowing medication in patients with AD and the relationships between these impairments, insight, and overall cognition. METHODS: Semistructured in-home interviews were conducted with 48 patients with very mild to moderate AD and 102 family caregivers of patients with mild to severe AD recruited from the Memory Disorders Clinic of an AD center. The interview measured performance on the decision-making abilities and three expert psychiatrists' judgment of competency based on their independent review of the patient interviews. RESULTS: There was considerable variation in patients' performance on the measures of decision-making abilities. Three expert raters found 19 of 48 (40%) of the subjects competent. Competent patients were more likely to show awareness of their symptoms, prognosis, and diagnosis. A sensitivity analysis suggests that a MMSE score is helpful in discriminating capacity from incapacity only when below 19 or above 23. CONCLUSIONS: Persons with mild to moderate Alzheimer disease (AD) have notable impairments in their ability to make an AD treatment decision, especially persons with moderate AD and persons who lack awareness of symptoms, prognosis, or diagnosis. PMID- 15883311 TI - MRI predictors of risk of incident Alzheimer disease: a longitudinal study. AB - OBJECTIVE: To determine if baseline entorhinal and hippocampal volumes and their rate of atrophy could predict the risk of incident Alzheimer disease (AD). METHODS: The authors used proportional odds models to assess the relationship between entorhinal and hippocampal size and risk of incident AD among 58 nondemented elderly people. All participants were followed with annual clinical evaluations and structural MRI scans for up to 5 years (baseline and 5 years of follow-up). At baseline, 23 of 58 participants received a diagnosis of amnestic mild cognitive impairment (MCI) and 35 of 58 were healthy control subjects with no cognitive impairment. Structural MRI scans were acquired with a T1-weighted three-dimensional spoiled gradient-recalled echo pulse sequence in a 1.5 T scanner. Entorhinal and hippocampal volumes were derived from 1.6-mm gapless coronal images reformatted to be perpendicular to the long axis of the hippocampus and were normalized by dividing with intracranial volume. RESULTS: Fourteen of 58 nondemented participants developed AD during the follow-up period. Initial diagnosis of MCI was a significant predictor of incident AD. In addition, both baseline entorhinal volume and its slope of decline were independent predictors of incident AD, but initial hippocampal size and its rate of decline were not, after controlling for entorhinal volume. CONCLUSION: In nondemented individuals, entorhinal cortex atrophy is associated with risk of Alzheimer disease. PMID- 15883312 TI - Incidence and etiology of dementia in a large elderly Italian population. AB - OBJECTIVE: To estimate age- and sex-specific incidence of dementia, Alzheimer disease (AD), and vascular dementia (VaD) in the Conselice Study of Brain Aging, an Italian prospective population-based study, and to assess whether poor education is a risk factor for dementia. METHODS: In 1999 to 2000, the baseline study identified a dementia-free cohort of 937 subjects aged 65 years and older who were reexamined in 2003 to 2004 using a two-phase procedure. RESULTS: Information was obtained for 91% of the subjects at risk; 115 incident cases of dementia were identified. Incidence rates per 1,000 person-years were 37.8 (95% CI = 30.0 to 47.7) for dementia, 23.8 (95% CI = 17.3 to 31.7) for AD, and 11.0 (95% CI = 7.2 to 16.9) for VaD. This translates into more than 400,000 new cases of dementia expected per year in Italy. Increasing age was an independent risk factor for both AD and VaD. Poor education was an independent risk factor for AD but not VaD. Sex did not affect dementia risk. CONCLUSIONS: In this Italian population-based cohort, incidence of dementia increased with age, and Alzheimer disease (AD) was the most frequent type of dementia. Poor education was associated with a higher risk of AD. Our incidence rates are higher than previously reported in Italy, and provide new estimates for projection of future burden of disease in Italy. PMID- 15883313 TI - APOE genotype, cholesterol level, lipid-lowering treatment, and dementia: the Three-City Study. AB - OBJECTIVE: To examine the association of plasma cholesterol levels, lipid lowering agent (LLA) intake, and APOE genotype with dementia prevalence. METHODS: The Three-City Study is a population-based cohort of 9,294 subjects selected from the electoral rolls of three French cities (Bordeaux, Dijon, Montpellier). Baseline examination included extensive assessment of exposure to vascular risk factors (including cholesterol levels and LLA use [statin or fibrate]) and clinical diagnosis of dementia. RESULTS: Two percent of participants were demented at baseline. Overall 32.4% of participants had hyperlipidemia, and 15.6% were prescribed statins and 13.7% fibrates. After adjusting for age, gender, education level, and study center, the odds ratio (OR) for dementia was observed to be lower among LLA users (OR = 0.61, 95% CI = 0.41 to 0.91) compared with subjects taking no LLAs. There was no differential effect between statin and fibrate users. The odds for dementia were increased in subjects with hyperlipidemia (OR = 1.43, 95% CI = 1.03 to 1.99). Further adjustment for potential confounders did not modify these associations. In addition, the association between LLA intake and dementia was not modified by APOE genotype, whereas hyperlipidemia was significantly associated with increased dementia prevalence only in non-epsilon4 carriers and non-Alzheimer disease cases. Finally, in participants taking LLAs, the odds for dementia were decreased only in those having normal lipid levels. CONCLUSIONS: This observational study provides further evidence that lipid-lowering agents are associated with decreased risk of dementia, whereas hyperlipidemia is associated with increased odds for non-Alzheimer-disease-type dementia. These effects appear to be independent of all major potential confounders. PMID- 15883314 TI - Classification of vascular dementia in the Cardiovascular Health Study Cognition Study. AB - OBJECTIVE: To describe the diagnostic classification of subjects with incident vascular dementia (VaD) participating in the Cardiovascular Health Study (CHS) Cognition Study. METHODS: The CHS classified 480 incident cases between 1994 and 1999 among 3,608 CHS participants who had brain MRI in 1992 through 1994 and in 1997 through 1998. The patients were diagnosed before and after reviewing the brain MRI. RESULTS: The pre-MRI classification showed that 52 participants had VaD and 76 had both Alzheimer disease (AD) and VaD. The post-MRI classification showed that the Diagnostic and Statistical Manual of Mental Disorders (4th ed.; DSM-IV) criteria classified 61 subjects as having VaD, the National Institute of Neurological Disorders and Stroke-Association Internationale pour la Recherche et l'Enseignement en Neurosciences (NINDS-AIREN) criteria classified 43 subjects as having probable VaD and 10 as possible VaD, and the State of California Alzheimer's Disease Diagnostic and Treatment Center (ADDTC) criteria classified 117 as having probable VaD and 96 as possible. The combination of the ADDTC and National Institute of Neurological and Communication Disorders and Stroke Alzheimer's Disease and Related Disorders Association criteria was used to examine the spectrum of vascular disease in dementia. The dementia was attributable to only vascular factors in 56 cases (probable VaD); VaD coexisted with AD in 61 cases, although the VaD component was the leading cause of dementia (probable VaD with AD); AD was the leading cause of dementia in 61 cases (possible VaD and probable AD); and in 29 cases, it was not clear that either AD or VaD was the primary diagnosis (possible AD and possible VaD). CONCLUSIONS: None of the clinical criteria for VaD identified the same group of subjects. The diagnosis of vascular dementia is difficult in epidemiologic studies because poststroke dementia can be due to Alzheimer disease (AD) and evidence of vascular disease can be found in the MRI of dementia cases without clinical strokes. Whether the clinical progression is related to AD pathology or vascular disease is difficult to establish. PMID- 15883315 TI - Determinants of vascular dementia in the Cardiovascular Health Cognition Study. AB - OBJECTIVE: The authors evaluated 3,375 participants without dementia at the time of MRI in 1991 to 1994 over 5.7 years for incident dementia and type of dementia. METHODS: Incidence of and risk factors for vascular dementia (VaD) were measured using both pre-MRI and modified State of California Alzheimer's Disease Diagnostic and Treatment Centers (ADDTC) post-MRI review and further classified Alzheimer disease (AD) by the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA) criteria. RESULTS: Approximately 44% (213) of 480 incident dementia cases were classified as possible or probable VaD by ADDTC. The incidence of VaD increased with age and was greater in blacks than whites. Risk factors for VaD included age, Modified Mini-Mental State Examination, high white matter grade, number of MRI infarcts, ventricular size, and history of stroke. CONCLUSIONS: Vascular disease in the brain is prevalent among incident dementia cases. There is a substantial overlap between cases classified as Alzheimer disease by Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association and vascular dementia (VaD) by modified State of California Alzheimer's Disease Diagnostic and Treatment Centers criteria. The substantial contribution of vascular disease would be missed without inclusion of MRI. Treatment of risk factors for VaD could have an important impact on incidence of dementia. PMID- 15883316 TI - Clinical effects of Abeta immunization (AN1792) in patients with AD in an interrupted trial. AB - BACKGROUND: AN1792 (beta-amyloid [Abeta]1-42) immunization reduces Abeta plaque burden and preserves cognitive function in APP transgenic mice. The authors report the results of a phase IIa immunotherapy trial of AN1792(QS-21) in patients with mild to moderate Alzheimer disease (AD) that was interrupted because of meningoencephalitis in 6% of immunized patients. METHODS: This randomized, multicenter, placebo-controlled, double-blind trial of IM AN1792 225 microg plus the adjuvant QS-21 50 microg (300 patients) and saline (72 patients) included patients aged 50 to 85 years with probable AD, Mini-Mental State Examination (MMSE) 15 to 26. Injections were planned for months 0, 1, 3, 6, 9, and 12. Safety and tolerability were evaluated, and pilot efficacy (AD Assessment Scale-Cognitive Subscale [ADAS-Cog], MRI, neuropsychological test battery [NTB], CSF tau, and Abeta42) was assessed in anti-AN1792 antibody responder patients (immunoglobulin G titer > or = 1:2,200). RESULTS: Following reports of meningoencephalitis (overall 18/300 [6%]), immunization was stopped after one (2 patients), two (274 patients), or three (24 patients) injections. Of the 300 AN1792(QS-21)-treated patients, 59 (19.7%) developed the predetermined antibody response. Double-blind assessments were maintained for 12 months. No significant differences were found between antibody responder and placebo groups for ADAS Cog, Disability Assessment for Dementia, Clinical Dementia Rating, MMSE, or Clinical Global Impression of Change, but analyses of the z-score composite across the NTB revealed differences favoring antibody responders (0.03 +/- 0.37 vs -0.20 +/- 0.45; p = 0.020). In the small subset of subjects who had CSF examinations, CSF tau was decreased in antibody responders (n = 11) vs placebo subjects (n = 10; p < 0.001). CONCLUSION: Although interrupted, this trial provides an indication that Abeta immunotherapy may be useful in Alzheimer disease. PMID- 15883317 TI - Effects of Abeta immunization (AN1792) on MRI measures of cerebral volume in Alzheimer disease. AB - BACKGROUND: Alzheimer disease (AD) is characterized by progressive cerebral atrophy that may be measured using MRI. Reported are MRI findings of a Phase IIa immunotherapy trial in AD prematurely terminated owing to meningoencephalitis in a subset of patients. OBJECTIVE: To assess cerebral volume changes in patients immunized with AN1792 (beta-amyloid [Abeta] 1 to 42) who were antibody responders (anti-AN1792 IgG titer of > or =1:2,200) compared with placebo patients. METHODS: This randomized, multicenter, placebo-controlled, double-blind trial of AN1792 225 mug plus QS-21 50 mug included 372 patients with probable AD. Patients received one to three injections of AN1792/QS-21 or saline and were assessed for 12 months. Volumetric MRI was performed pre dose and at month 12 or early termination. Brain, ventricular, and hippocampal volume changes were measured from registered scan pairs. RESULTS: Two hundred eighty-eight patients had paired scans (mean interval 10.9 months). Antibody responders (n = 45) had greater brain volume decrease (3.12 +/- 1.98 vs 2.04 +/- 1.74%; p = 0.007), greater ventricular enlargement as a percentage of baseline brain volume (1.10 +/- 0.75 vs 0.48 +/- 0.40%; p < 0.001), and a nonsignificant greater hippocampal volume decrease (3.78 +/- 2.63 vs 2.86 +/- 3.19%; p = 0.124) than placebo patients (n = 57). Increased losses in brain volume were not reflected in worsening cognitive performance; a composite z score across a Neuropsychological Test Battery showed differences favoring antibody responders over placebo (0.03 +/- 0.39 vs -0.24 +/- 0.45; p = 0.008). CONCLUSIONS: A dissociation between brain volume loss and cognitive function was observed in AN1792/QS-21 antibody responders. The reasons for this remain unclear but include the possibility that volume changes were due to amyloid removal and associated cerebral fluid shifts. PMID- 15883318 TI - Headache, cerebrovascular symptoms, and stroke: the Atherosclerosis Risk in Communities Study. AB - OBJECTIVE: To evaluate the occurrence of stroke/TIA symptoms and ischemic stroke events among those with a lifetime history of migraine or other headaches with some migraine features in a biracial cohort of older adults. METHODS: Participants were 12,750 African-American and white men and women from the Atherosclerosis Risk in Communities Study (1993 to 1995). The participants were queried about their lifetime headache history and characterized using modified International Headache Society diagnostic criteria. Stroke/TIA symptoms were classified using a computerized diagnostic algorithm, and ischemic stroke events were identified and validated using medical records. Multivariate logistic regression was used to assess the relationship between headache types and stroke/TIA symptoms and ischemic stroke events. RESULTS: Migraine with aura was strongly associated with stroke symptoms (odds ratio [OR] 5.46, 95% CI: 3.64 to 8.18), TIA symptoms (OR 4.28, 95% CI: 3.02 to 6.08), and verified ischemic stroke events (OR 2.81, 95% CI: 1.60 to 4.92). Similarly, other headaches with aura were significantly associated with stroke symptoms (OR 3.68, 95% CI: 2.26 to 5.99) and TIA symptoms (OR 4.53, 95% CI: 3.08 to 6.67). In contrast, the associations for migraine without aura and other headaches without aura were not as consistent or robust. CONCLUSIONS: Migraines and other headaches, particularly those accompanied by aura, were associated with an increased occurrence of stroke/TIA symptoms and ischemic stroke events. PMID- 15883319 TI - A novel mutation (K317M) in the MAPT gene causes FTDP and motor neuron disease. AB - BACKGROUND: Frontotemporal dementia with parkinsonism is often linked to chromosome 17 and is related to mutations in the MAPT gene. In some families the genetic basis is still unknown. The authors report two pedigrees with FTDP-17 harboring a novel mutation (K317M) in exon 11 in the MAPT gene. METHODS: The authors identified two apparently unrelated pedigrees with an autosomal dominant neurodegenerative condition. Thirteen patients were examined and eight autopsies were performed. RESULTS: Mean age at onset was 48 years. Mean disease duration was 6 years. Dysarthria often heralded the disease. All cases had parkinsonism and pyramidalism and half of them had amyotrophy. Behavioral or personality changes were not a prominent feature. Cognitive decline appeared late in the evolution. Neuropathologically, a massive degeneration of the substantia nigra without Lewy bodies was a constant finding. A variable degree of frontotemporal atrophy was found. Corticospinal tract degeneration and anterior horn neuron loss were present in six of seven autopsies in which the spinal cord was examined. An extensive deposition of abnormal tau protein in a mixed pattern (neuronal, glial) was observed. Pick's bodies were not seen. Biochemical analysis of tau revealed two bands of 64 and 68 kDa. CONCLUSION: Genetic analysis revealed the same novel mutation (K317M) in exon 11 of the MAPT gene in both pedigrees. A common haplotype between members of the two pedigrees suggests that they belong to the same family. PMID- 15883320 TI - Multiple cervical artery dissections. PMID- 15883321 TI - Mortality from Creutzfeldt-Jakob disease and related disorders in Europe, Australia, and Canada. AB - BACKGROUND: An international study of the epidemiologic characteristics of Creutzfeldt-Jakob disease (CJD) was established in 1993 and included national registries in France, Germany, Italy, the Netherlands, Slovakia, and the United Kingdom. In 1997, the study was extended to Australia, Austria, Canada, Spain, and Switzerland. METHODS: Data were pooled from all participating countries for the years 1993 to 2002 and included deaths from definite or probable CJD of all etiologic subtypes. RESULTS: Four thousand four hundred forty-one cases were available for analysis and included 3,720 cases of sporadic CJD, 455 genetic cases, 138 iatrogenic cases, and 128 variant cases. The overall annual mortality rate between 1999 and 2002 was 1.67 per million for all cases and 1.39 per million for sporadic CJD. Mortality rates were similar in all countries. There was heterogeneity in the distribution of cases by etiologic subtype with an excess of genetic cases in Italy and Slovakia, of iatrogenic cases in France and the UK, and of variant CJD in the UK. CONCLUSIONS: This study has established overall epidemiologic characteristics for Creutzfeldt-Jakob disease (CJD) of all types in a multinational population-based study. Intercountry comparisons did not suggest any relative change in the characteristics of sporadic CJD in the United Kingdom, and the evidence in this study does not suggest the occurrence of a novel form of human bovine spongiform encephalopathy infection other than variant CJD. However, this remains a possibility, and countries currently unaffected by variant CJD may yet have cases. PMID- 15883322 TI - High incidence of genetic human transmissible spongiform encephalopathies in Italy. AB - OBJECTIVE: To assess the incidence and mortality rates of genetic transmissible spongiform encephalopathy (TSE) diseases in Italy. METHODS: The authors have sequenced the prion protein gene (PRNP) in 643 patients referred to the Italian Registry of Creutzfeldt-Jakob disease (CJD) and related disorders between 1993 and 2002. Crude age- and sex-specific incidence and mortality rates were calculated. Differences in morbidity from genetic TSE diseases in the 20 Italian regions were assessed by the standardized morbidity ratio (SMR). RESULTS: A total of 130 cases were classified as genetic TSE diseases with a mean yearly incidence rate of 0.28 cases per million people. Genetic TSE diseases represent 17.7% of all TSE diseases, including sporadic, iatrogenic, and variant CJD. The most frequent mutation was the V210I (n = 54), and the second most common the E200K (n = 42). Mortality rates for genetic TSE diseases did not increase in any of the age groups under examination over the 10 years of surveillance. The analysis of regional distribution of genetic cases by place of birth revealed that in Campania and Calabria regions the number of genetic TSE cases was higher than in other regions. CONCLUSIONS: In Italy the incidence of genetic transmissible spongiform encephalopathy (TSE) diseases is the second highest among European countries. Genetic analysis is important for a correct classification of patients with TSE. PMID- 15883323 TI - Parkinson disease, brain volumes, and subthalamic nucleus stimulation. AB - BACKGROUND: High-frequency stimulation of the subthalamic nucleus (STN) is an effective treatment for advanced Parkinson disease (PD). The clinical and preoperative predictive factors of the best postoperative outcome have been identified. Radiologic predictive factors were investigated. METHODS: Forty patients with PD underwent surgery for bilateral STN stimulation. MRI was performed in stereotactic conditions before surgery. Brain parenchyma, caudate nucleus, putamen, pallidum, and red nucleus volumes and the surface of the mesencephalon were measured and normalized as percentages of the intracranial volume. Clinical evaluation was performed 1 month before and 6 months after surgery. RESULTS: The normalized brain parenchyma volume was lower in patients who were older and had a longer disease duration or a lower frontal score and was not predictive of the postoperative outcome. The residual scores for activities of daily living and parkinsonian motor disability were higher in patients with a smaller normalized mesencephalon. The normalized caudate nucleus volume was predictive of the pre- and postoperative levodopa-equivalent dosage. CONCLUSIONS: Brain atrophy is not an exclusion criterion for neurosurgery, indicating that patients' neurologic, psychiatric, and neuropsychological characteristics are the best predictive factors for neurosurgery. The fact that a smaller normalized mesencephalon surface was associated with a lower beneficial effect of the subthalamic nucleus stimulation on the parkinsonian motor disability suggests that the normalized mesencephalon surface is a predictive factor of the postoperative outcome. PMID- 15883324 TI - Antecedent infections in Fisher syndrome: a common pathogenesis of molecular mimicry. AB - OBJECTIVE: To assess the production mechanism of anti-GQ1b autoantibody in Fisher syndrome (FS). METHODS: The authors conducted a prospective case-control serologic study of five antecedent infections (Campylobacter jejuni, cytomegalovirus, Epstein-Barr virus, Mycoplasma pneumoniae, and Haemophilus influenzae) in 73 patients with FS and 73 sex- and age-matched hospital controls (HCs). Serologic evidence in FS patients of C. jejuni (21%) and H. influenzae (8%) infections was present significantly more often than in the HCs. None of the five pathogens examined was found in the 49 (67%) patients with FS. Anti-GQ1b IgG antibody was detected in most FS patients infected with C. jejuni or H. influenzae. Mass spectrometry analysis identified a C. jejuni strain (CF93-6) carrying a GT1a-like lipo-oligosaccharide (LOS) that had been isolated from an FS patient. Immunization of complex ganglioside-lacking knockout mice with the GT1a like LOS generated IgG class monoclonal antibodies (mAbs) that reacted with GQ1b and GT1a. Thin-layer chromatography with immunostaining showed that anti-GQ1b mAb bound to the C. jejuni LOS (50% of the 20 FS-related strains) more commonly than in the Guillain-Barre syndrome (GBS)-related (7% of 70) or enteritis-related (20% of 65) strains. Anti-GM1 and anti-GD1a mAbs also reacted with the LOS from some FS-related strains (both 20%), but binding frequencies were higher in the GBS related strains (74 and 57%). The GQ1b epitope was detected in 4 (40%) of the 10 FS-related H. influenzae strains but was absent in strains from patients with GBS (n = 4) and uncomplicated respiratory infections (n = 10). CONCLUSIONS: C. jejuni and H. influenzae are related to Fisher syndrome (FS) development, and production of anti-GQ1b autoantibody is mediated by the GQ1b-mimicking lipo-oligosaccharides on those bacteria. The causative agents remain unclear in the majority of patients with FS. PMID- 15883325 TI - IV thrombolysis in patients with acute stroke due to spontaneous carotid dissection. AB - The authors reviewed the histories of 33 patients (ages 44 to 50 years) treated with IV thrombolysis for acute stroke due to spontaneous cervical carotid artery dissection. Median NIH Stroke Scale (NIHSS) score on admission was 15. No new or worsened local signs, subarachnoid hemorrhage, pseudoaneurysm formation, or rupture of the cervical ICA were observed. At 3 months, median NIHSS was 7 and median modified Rankin Scale (mRS) 2.5; mRS < or = 2 was observed in 17 patients. PMID- 15883326 TI - Multidirectional transpositions suggesting pathologic approach behavior after frontal stroke. AB - The authors report a patient with a right frontal stroke who, despite the ability to draw clocks accurately from memory, translocated numbers on the inferior half of the dial to the superior half when copying a clock. In further graphic and verbal clock reproduction tasks, transpositions were always directed toward the model but disappeared in a delayed copying task. These findings appear to reflect an intentional disorder characterized by pathologic approach behavior. PMID- 15883327 TI - Phosphorylated 14-3-3zeta protein in the CSF of neuroleptic-treated patients. AB - The authors describe 12 neuroleptic-treated patients with dementia of various etiologies who showed CSF elevation of phosphorylated 14-3-3zeta and normal tau protein levels. This contrasted with elevated amounts of 14-3-3 gamma, epsilon, and unphosphorylated zeta coupled to high tau protein levels in Creutzfeldt-Jakob disease and negative 14-3-3 assay in drug-free patients with dementia. Characterization of CSF 14-3-3 isoforms and determination of tau protein level can help to distinguish different etiologies of dementia. PMID- 15883328 TI - The natural history of Aicardi-Goutieres syndrome: follow-up of 11 Italian patients. AB - Described are the outcomes of 11 Italian patients with Aicardi-Goutieres syndrome. Neurologic symptoms progressed in the first year of life and stabilized by the end of the second year in 10 patients. White matter abnormalities remained stable; cerebral atrophy was stable in four patients and progressive in two. Calcifications increased (in number and size) in two of six patients. Serial CSF and serum interferon-alpha measurements (three patients) showed reduced CSF interferon-alpha levels. PMID- 15883329 TI - Ventriculoperitoneal shunt in patients with leptomeningeal metastasis. AB - The authors reviewed 37 patients with leptomeningeal metastasis (LM) who required a ventriculoperitoneal shunt (VP shunt) for management of intracranial hypertension. Improvement was seen in 27 (77%) patients; subdural hematoma developed in one and shunt malfunction in three. Median overall survival was 2 months (range 2 days to 3.6 years) after VP shunt placement, but there was no procedure-related mortality. The prognosis of LM remained poor, but VP shunt can be an effective palliative tool when required. PMID- 15883330 TI - Patterns of inheritance in familial ALS. AB - We investigated 185 families with ALS for evidence of anticipation and mitochondrial inheritance. Although initial analysis demonstrated significant anticipation of age at death between generations in patients with familial ALS, further analysis demonstrated features of regression to the mean, suggesting that the perceived differences are the result of bias. In addition, there was no evidence of an effect of preferential maternal inheritance, which would have supported transmission of mitochondrial DNA mutations. PMID- 15883331 TI - Kernohan notch lesion after spinal tap. PMID- 15883332 TI - Anti-sulfatide antibodies in HIV-infected individuals with sensory neuropathy. AB - We used ELISA to estimate the prevalence of anti-sulfatide antibodies in HIV infected individuals with distal sensory neuropathy (DSP) and compared the results with the prevalence in HIV-infected individuals without DSP and in individuals with neuropathy who are not infected with HIV. We found that 36% of HIV+/DSP+ individuals had immunoglobulin (Ig) G anti-sulfatide antibody titers greater than 1,500, whereas IgG anti-sulfatide antibodies were rarely found in HIV+/DSP- or HIV-/DSP+ patients. PMID- 15883333 TI - Hemimegalencephaly and tuberous sclerosis complex. PMID- 15883334 TI - LGMD2I presenting with a characteristic Duchenne or Becker muscular dystrophy phenotype. AB - LGMD type 2I, caused by mutations in the fukutin-related protein, is a common form of LGMD. The phenotype resembles Duchenne/Becker muscular dystrophy. A point mutation, L276I has been found in all patients with LGMD2I studied so far. The authors screened for this mutation in 102 sporadic cases of Duchenne/Becker mutation-negative patients and found 13 patients with LGMD2I. PMID- 15883335 TI - Myopathy with skeletal asymmetry and hemidiaphragm elevation is caused by myotubularin mutations. AB - The authors report two families with a myopathy phenotype affecting only women, marked by asymmetric weakness, skeletal asymmetry, and an elevated hemidiaphragm. One family had a mutation in a stop codon in exon 9 of the myotubularin gene, and the other had a splice site mutation in exon 13. Both families had manifesting and nonmanifesting carriers. Skewed X-inactivation appeared to explain the clinical manifestations in only one of the two families. PMID- 15883336 TI - Long-term results of tacrolimus in cyclosporine- and prednisone-dependent myasthenia gravis. AB - Seventy-nine patients with cyclosporine- and prednisone-dependent myasthenia gravis (MG) after thymectomy received tacrolimus for a mean of 2.5 +/- 0.8 years. Prednisone was withdrawn in all but two patients. Anti-acetylcholine antibodies and MG score for disease severity decreased significantly and muscular strength increased by 39%. Complete stable remission was achieved in 5% of patients and pharmacologic remission in 87.3%. All patients resumed full activities of daily living. PMID- 15883337 TI - Effects of chronic lead exposure on 1H MRS of hippocampus and frontal lobes in children. AB - The authors performed IQ testing and magnetic resonance spectroscopy on six lead exposed and six control children. Levels of N-acetyl aspartate (neuronal density and mitochondrial metabolism), creatine + phosphocreatine (phosphate metabolism), and choline (membrane turnover) were decreased in four brain regions (left and right frontal, left and right hippocampus) in lead-exposed children vs controls. The reductions were right frontal > left frontal > hippocampus but were the same bilaterally in the hippocampus. PMID- 15883338 TI - A writing device improves writing in primary writing tremor. AB - Primary writing tremor (PWT) is task-specific and interferes with handwriting. Several reports have shown a beneficial response of this disorder to stereotactic functional neurosurgery. Significant improvement with a writing device was demonstrated with blind rating of handwriting and spiral drawing samples collected before and during its use in nine patients with PWT, suggesting that this therapeutic modality should be tried before considering chronic pharmacotherapy or functional neurosurgery. PMID- 15883339 TI - Identification of candidates for epilepsy surgery in patients with tuberous sclerosis. AB - The authors reviewed preoperative MRI and EEG findings in relation to postsurgical outcome in 17 patients with refractory epilepsy due to tuberous sclerosis complex (TSC). Resecting concordant MRI (main tuber) and EEG abnormalities offered seizure freedom (8/9, 89%; median follow-up 25 months) comparable to other focal etiologies. Patients with nonconcordant MRI and EEG findings did less well (3/8, 38%, seizure free; p = 0.027, OR = 13). PMID- 15883340 TI - High-dose creatine therapy for Huntington disease: a 2-year clinical and MRS study. PMID- 15883341 TI - Lack of hemispheric localizing value of the palmomental reflex. PMID- 15883342 TI - Post-traumatic epidermoid cyst presenting with headache. PMID- 15883343 TI - Screening for neurofilament inclusion disease using alpha-internexin immunohistochemistry. PMID- 15883344 TI - Perisylvian polymicrogyria in Landau-Kleffner syndrome. PMID- 15883345 TI - Atypical PML leading to a diagnosis of common variable immunodeficiency. PMID- 15883346 TI - Personal history: code team. PMID- 15883347 TI - Ulnar neuropathy at the elbow: follow-up and prognostic factors determining outcome. PMID- 15883348 TI - Does acute occlusion of the carotid T invariably have a poor outcome? PMID- 15883349 TI - An AUPN/ANA survey of department leader opinions on the health of US academic neurology. PMID- 15883350 TI - The occurrence of Guillain-Barre syndrome within families. PMID- 15883351 TI - Single-fiber EMG in familial hemiplegic migraine. PMID- 15883352 TI - Rules for rehabilitation: an agenda for research. AB - Rehabilitation services have grown tremendously in the United States over the past 2 decades. Rules originally designed to guide Medicare reimbursement policies have had substantial effects in shaping the design of clinical services. This article traces the development of the most significant federal rules regarding rehabilitation, outlines the existing empirical evidence to support these rules, and discusses an agenda for research to improve the evidence for future policy development. PMID- 15883353 TI - Line bisection and rebisection: the crossover effect of space location. AB - OBJECTIVE: To investigate the relationship between the bisection test and the severity of behavioral hemineglect and to verify if this test can predict the behavioral hemineglect. METHODS: Thirty stroke patients with left hemiparesis were divided into 4 groups according to the Catherine Bergego Scale, which assessed the behavioral hemineglect: severe unilateral neglect (UN), moderate UN, mild UN, and lack of UN. Eleven healthy subjects served as age-matched control subjects. In the bisection test, 18 lines were presented on the left, middle, and right of an A4 paper, respectively. The subjects were asked to place a short cross mark in the exact middle point of each line on the paper using their right hand. The middle 6 lines in the above bisection test were extracted on another sheet of A4 paper for the rebisection test. The subjects were asked to divide a line into 4 segments by successive bisections. The proportion of the right part to the length of line for bisecting was calculated. RESULTS: In the bisection test, the main effect of space was significant in every group except the mild neglect group. The crossover effect of space location was found in the severe UN group, the group without UN, and the controls. In the severe UN group, the patients bisected the left and middle lines with rightward bias (<50%) but bisected the right lines with leftward bias (>50%). In the group without UN and the controls, the subjects bisected the left lines with leftward bias (>50%) but bisected the middle and right lines with rightward bias (<50%). Almost the same results were seen in the rebisection test. CONCLUSIONS: This study showed that if the spatial crossover effect occurred in the right space condition, it was strongly supported that this patient had moderate to severe behavioral hemineglect. The crossover effect of the space location was explained by a new model. PMID- 15883354 TI - Learning motor sequences with and without knowledge of governing rules. AB - OBJECTIVE: To investigate the behavioral and neural effects of rule-based knowledge on motor sequence learning. METHODS: The authors developed a novel 2 dimensional variant of the serial reaction time (SRT) task to test the effect of prior, verbalizable rule knowledge on motor learning behavior. To examine neurophysiological effects, they also performed functional magnetic resonance imaging on a small cohort of subjects while performing the same task. RESULTS: Behavioral data demonstrated that instruction on sequence-governing rules enhanced behavioral performance in both learning magnitudes and rates. The neuroimaging data revealed substantially different, but partially overlapping, learning-related activation patterns with and without prior rule instruction. Direct comparison of these 2 conditions revealed significantly different involvement of bilateral superior and anterior prefrontal cortex (Brodmann areas 8 and 10, respectively), right superior temporal cortex (BA 38/21), and left cerebellum. CONCLUSIONS: These behavioral findings demonstrate an advantage of teaching governing rules prior to 2D-SRT task performance. While these neuroimaging findings remain to be replicated in a larger cohort of subjects, results suggest that substantially different-though partially overlapping-brain regions subserve learning in these 2 rehabilitation-relevant conditions. Thus, appropriate choice of pretraining may benefit, for example, rehabilitation populations, at least in motor skill acquisition that requires sequencing. PMID- 15883355 TI - Speed and temporal-distance adaptations during treadmill and overground walking following stroke. AB - OBJECTIVE: To compare the maximum gait speed of stroke subjects attained during treadmill and overground in stroke subjects and to identify the temporal-distance determinants of the maximal gait speed. METHODS: Ten individuals with hemiparetic gait deficits and whose walking speeds ranged between 0.24 m/s and 0.82 m/s participated. Five healthy age-matched controls were also tested to provide comparative data for the gait speed transfer between the 2 modes of locomotion. Following a brief habituation process to walking on the treadmill, subjects were tested while walking at comfortable and maximal speeds on the treadmill and overground, in a random order. Main Outcome Measure. Self-selected comfortable and maximum gait speed and temporal-distance factors were acquired using a 6 camera Vicontrade mark motion analysis system and compared between treadmill and overground walking at a similar speed. RESULTS: Overground walking resulted in higher maximal speeds (P < 0.001), greater stride lengths (P < 0.001), and a lower cadence (P < 0.02), as compared to tread-mill. The comfortable gait speed and the maximum stride length proved to be strong determinants for the maximal speed on both modes of locomotion (P < 0.01), but the maximum cadence was correlated to maximum speed only for overground locomotion (P < 0.05). CONCLUSIONS: Stroke subjects walked slower on the treadmill as compared to overground. They also used a different strategy to increase gait speed, relying mostly on increasing the stride length during treadmill ambulation. PMID- 15883356 TI - Evidence for cognitive processes involved in the control of steady state of walking in healthy subjects and after cerebral damage. AB - OBJECTIVE: The involvement of cognitive processes in the control of walking at steady state was studied in 10 healthy subjects and 18 subjects after unilateral vascular brain damage. A dual task paradigm was used to compare the performance level of a probe reaction time (RT) in sitting (simple task) and during standing or walking on a treadmill (dual task conditions). In this latter condition, RTs were classified and analyzed in relation to the different phases of the gait cycle. RESULTS: The results show a marked increase in RT while walking compared to sitting and standing only in stroke subjects. Specific changes in RTs related to the gait cycle phases were observed in both healthy subjects and those after brain damage. It is concluded that walking at steady state is attentionally demanding. The phase-dependent modulations of the RTs suggest that cognitive processes may play a role in the control of the step cycle. The increase of attentional demand during walking in subjects who had suffered a stroke varies, depending on severity of impairments of walking but also on a reduced general attentional capacity. The dual task paradigm provides a sensitive tool in the assessment of walking ability in stroke subjects. PMID- 15883357 TI - Relationship between ischemic lesion volume and functional status in the 2nd week after middle cerebral artery stroke. AB - OBJECTIVE: To examine the relationship between the volume of the middle cerebral artery stroke lesion and functional status in the subacute phase of stroke. METHODS: Infarctvolumes of 94 patients with a 1st middle cerebral artery stroke assessed on conventional MRI scans obtained in the 2nd week poststroke were related to a clinical measure of stroke severity (National Institutes of Health Stroke Scale [NIHSS]) and to functional status: motor impairment (Motricity Index [MI]) and limitation in activities (Barthel Index [BI] and modified Rankin Scale). Separate correlations were computed for patients with large (>30 ml) and small (30 ml) lesions, and to investigate the influence of lesion location on the relationship between volume and functional status, correlations were computed for patients with left and right hemisphere lesions and for patients with cortical and subcortical lesions. RESULTS: Lesion volume correlated strongly with NIHSS scores (R = 0.61) and moderately with the patient's functional status (MI [R between -0.42 and - 0.49], BI [R = -0.43], and Modified Rankin Scale [R = 0.45]). Right hemisphere lesions and cortical lesions had a stronger correlation with functional status. In patients with small lesion volumes (0-30 ml), no relationship between lesion volumes and functional status was seen at all. CONCLUSIONS: Lesion volume is moderately to strongly related to the functional status in the 2nd week poststroke. PMID- 15883358 TI - Stroke rehabilitation in Switzerland versus the United States: a preliminary comparison. AB - This article compares the structure and process of rehabilitation for stroke patients at 2 internationally recognized rehabilitation hospitals, Klinik Valens ("Valens") in Switzerland and the William Donald Schaeffer Rehabilitation Hospital at Kernan ("Kernan") in the United States. Although the patient mix, structure, and process of rehabilitation were similar in many regards, there were some important differences. Most notably, on average, patients at the U.S. hospital were discharged from rehabilitation at approximately the same day poststroke that rehabilitation began in Switzerland. Patients remained in an inpatient setting an average of 40 days longer in Switzerland (for the combination of acute care and rehabilitation) and had significantly higher levels of functioning at discharge when compared to their U.S. counterparts. The authors' findings suggest that Europe may offer opportunities for rehabilitation research that would be difficult to duplicate in the United States and highlight policy-relevant questions for future studies aimed at developing efficient managed care systems for stroke survivors. PMID- 15883359 TI - Noncholinergic excitatory actions of motoneurons in the neonatal mammalian spinal cord. AB - Mammalian spinal motoneurons are considered to be output elements of the spinal cord that generate exclusively cholinergic actions on Renshaw cells, their intraspinal synaptic targets. Here, we show that antidromic stimulation of motor axons evokes depolarizing monosynaptic potentials in Renshaw cells that are depressed, but not abolished, by cholinergic antagonists. This residual potential was abolished by 2-amino-5-phosphonovaleric acid and 6-cyano-7-nitroquinoxaline 2,3-dione. In the presence of cholinergic antagonists, motor axon stimulation triggered locomotor-like activity that was blocked by 2-amino-5-phosphonovaleric acid. Some cholinergic motoneuronal terminals on both Renshaw cells and motoneurons were enriched in glutamate, but none expressed vesicular glutamate transporters. Our results raise the possibility that motoneurons release an excitatory amino acid in addition to acetylcholine and that they may be more directly involved in the genesis of mammalian locomotion than previously believed. PMID- 15883360 TI - Chromosomal clustering of genes controlled by the aire transcription factor. AB - Autoimmune regulator (aire) is a transcription factor that controls the self reactivity of the T cell repertoire. Although previous results indicate that it exerts this function in part by promoting ectopic expression of a battery of peripheral-tissue antigens in epithelial cells of the thymic medulla, recent data argue for additional roles in negative selection of thymocytes by medullary cells. As one approach to exploring such roles, we performed computational analyses of microarray data on medullary RNA transcripts from aire-deficient versus wild-type mice, focusing on the genomic localization of aire-controlled genes. Our results highlight this molecule's transcriptional activating and silencing roles and reveal a significant degree of clustering of its target genes. On a local scale, aire-regulated clusters appeared punctate, with aire controlled and aire-independent genes often being interspersed. This pattern suggests that aire's action may not be a simple reflection of the wide action of a chromatin remodeling enzyme. Analysis of the identity of certain of the clustered genes was evocative of aire's potential roles in antigen presentation and the coordination of intrathymic cell migration: for example, major histocompatibility complex class I and class II gene products and certain chemokine genes are targets of aire-regulated transcription. PMID- 15883361 TI - Yeast genome-wide screen reveals dissimilar sets of host genes affecting replication of RNA viruses. AB - Viruses are devastating pathogens of humans, animals, and plants. To further our understanding of how viruses use the resources of infected cells, we systematically tested the yeast single-gene-knockout library for the effect of each host gene on the replication of tomato bushy stunt virus (TBSV), a positive strand RNA virus of plants. The genome-wide screen identified 96 host genes whose absence either reduced or increased the accumulation of the TBSV replicon. The identified genes are involved in the metabolism of nucleic acids, lipids, proteins, and other compounds and in protein targeting/transport. Comparison with published genome-wide screens reveals that the replication of TBSV and brome mosaic virus (BMV), which belongs to a different supergroup among plus-strand RNA viruses, is affected by vastly different yeast genes. Moreover, a set of yeast genes involved in vacuolar targeting of proteins and vesicle-mediated transport both affected replication of the TBSV replicon and enhanced the cytotoxicity of the Parkinson's disease-related alpha-synuclein when this protein was expressed in yeast. In addition, a set of host genes involved in ubiquitin-dependent protein catabolism affected both TBSV replication and the cytotoxicity of a mutant huntingtin protein, a candidate agent in Huntington's disease. This finding suggests that virus infection and disease-causing proteins might use or alter similar host pathways and may suggest connections between chronic diseases and prior virus infection. PMID- 15883362 TI - Perinatal omega-3 polyunsaturated fatty acid supply modifies brain zinc homeostasis during adulthood. AB - Dietary omega-3 polyunsaturated fatty acid (PUFA) influences the expression of a number of genes in the brain. Zinc transporter (ZnT) 3 has been identified as a putative transporter of zinc into synaptic vesicles of neurons and is found in brain areas such as hippocampus and cortex. Neuronal zinc is involved in the formation of amyloid plaques, a major characteristic of Alzheimer's disease. The present study evaluated the influence of dietary omega-3 PUFA on the expression of the ZnT3 gene in the brains of adult male Sprague-Dawley rats. The rats were raised and/or maintained on a control (CON) diet that contained omega-3 PUFA or a diet deficient (DEF) in omega-3 PUFA. ZnT3 gene expression was analyzed by using real-time PCR, free zinc in brain tissue was determined by zinquin staining, and total zinc concentrations in plasma and cerebrospinal fluid were determined by atomic absorption spectrophotometry. Compared with CON-raised animals, DEF-raised animals had increased expression of ZnT3 in the brain that was associated with an increased level of free zinc in the hippocampus. In addition, compared with CON raised animals, DEF-raised animals had decreased plasma zinc level. No difference in cerebrospinal fluid zinc level was observed. The results suggest that overexpression of ZnT3 due to a perinatal omega-3 PUFA deficiency caused abnormal zinc metabolism in the brain. Conceivably, the influence of dietary omega-3 PUFA on brain zinc metabolism could explain the observation made in population studies that the consumption of fish is associated with a reduced risk of dementia and Alzheimer's disease. PMID- 15883363 TI - Late-emigrating neural crest cells in the roof plate are restricted to a sensory fate by GDF7. AB - Lineage-tracing experiments have shown that some premigratory neural crest cells generate both sensory (S) and autonomic (A) derivatives, whereas others generate only S derivatives. Whether this lineage heterogeneity reflects random variation in a homogeneous population or an early sensory specification of some premigratory crest cells has not been clear. Using Cre recombinase-based fate mapping, we show that GDF7, which is exclusively expressed in the roof plate, marks neural crest cells with a 10-fold higher bias to the sensory lineage than those marked (at the same stage of development) by an inducible Wnt1-Cre, which is expressed more broadly in the dorsal neural tube. In vitro, GDF7 has potent sensory neuron-inducing activity. These data suggest that some premigratory crest cells are deterministically restricted to the S lineage and implicate GDF7 itself in this restriction process. PMID- 15883364 TI - Conservation and evolvability in regulatory networks: the evolution of ribosomal regulation in yeast. AB - Transcriptional modules of coregulated genes play a key role in regulatory networks. Comparative studies show that modules of coexpressed genes are conserved across taxa. However, little is known about the mechanisms underlying the evolution of module regulation. Here, we explore the evolution of cis regulatory programs associated with conserved modules by integrating expression profiles for two yeast species and sequence data for a total of 17 fungal genomes. We show that although the cis-elements accompanying certain conserved modules are strictly conserved, those of other conserved modules are remarkably diverged. In particular, we infer the evolutionary history of the regulatory program governing ribosomal modules. We show how a cis-element emerged concurrently in dozens of promoters of ribosomal protein genes, followed by the loss of a more ancient cis-element. We suggest that this formation of an intermediate redundant regulatory program allows conserved transcriptional modules to gradually switch from one regulatory mechanism to another while maintaining their functionality. Our work provides a general framework for the study of the dynamics of promoter evolution at the level of transcriptional modules and may help in understanding the evolvability and increased redundancy of transcriptional regulation in higher organisms. PMID- 15883365 TI - Prefrontal cortex and flexible cognitive control: rules without symbols. AB - Human cognitive control is uniquely flexible and has been shown to depend on prefrontal cortex (PFC). But exactly how the biological mechanisms of the PFC support flexible cognitive control remains a profound mystery. Existing theoretical models have posited powerful task-specific PFC representations, but not how these develop. We show how this can occur when a set of PFC-specific neural mechanisms interact with breadth of experience to self organize abstract rule-like PFC representations that support flexible generalization in novel tasks. The same model is shown to apply to benchmark PFC tasks (Stroop and Wisconsin card sorting), accurately simulating the behavior of neurologically intact and frontally damaged people. PMID- 15883366 TI - Laser-capture microdissection of plasma cells from subacute sclerosing panencephalitis brain reveals intrathecal disease-relevant antibodies. AB - Increased IgG and oligoclonal bands are found in cerebrospinal fluid of humans with chronic infectious CNS disease. Studies have shown that these oligoclonal bands are antibodies directed against the agent that causes disease. Laser capture microdissection was used to isolate individual CD38+ plasma cells from the brain of a patient with subacute sclerosing panencephalitis, and single-cell RT-PCR was used to analyze individual IgG heavy and light chains expressed by each cell. Based on overrepresented IgG sequences, we constructed functional recombinant antibodies (recombinant IgGs) and determined their specificities. Five of eight recombinant IgGs recognized measles virus, the cause of subacute sclerosing panencephalitis. These results demonstrate that overrepresented IgG sequences in postmortem brains can be used to produce functional recombinant antibodies that recognize their target antigens. This strategy can be used to identify disease-relevant antigens in CNS inflammatory diseases of unknown etiology. PMID- 15883367 TI - Genomic and proteomic comparisons between bacterial and archaeal genomes and related comparisons with the yeast and fly genomes. AB - Bacterial, archaeal, yeast, and fly genomes are compared with respect to predicted highly expressed (PHX) genes and several genomic properties. There is a striking difference in the status of PHX ribosomal protein (RP) genes where the archaeal genome generally encodes more RP genes and fewer PHX RPs compared with bacterial genomes. The increase in RPs in archaea and eukaryotes compared with that in bacteria may reflect a more complex set of interactions in archaea and eukaryotes in regulating translation, e.g., differences in structure requiring scaffolding of longer rRNA molecules, expanded interactions with the chaperone machinery, and, in eukaryotic interactions with endoplasmic reticulum components. The yeast genome is similar to fast-growing bacteria in PHX genes but also features several cytoskeletal genes, including actin and tropomyosin, and several signal transduction regulatory proteins from the 14.3.3 family. The most PHX genes of Drosophila encode cytoskeletal and exoskeletal proteins. We found that the preference of a microorganism for an anaerobic metabolism correlates with the number of PHX enzymes of the glycolysis pathway that well exceeds the number of PHX enzymes acting in the tricarboxylic acid cycle. Conversely, if the number of PHX enzymes of the tricarboxylic acid cycle well exceeds the PHX enzymes of glycolysis, an aerobic metabolism is preferred. Where the numbers are approximately commensurate, a facultative growth behavior prevails. PMID- 15883368 TI - Predicted highly expressed genes in archaeal genomes. AB - Based primarily on 16S rRNA sequence comparisons, life has been broadly divided into the three domains of Bacteria, Archaea, and Eukarya. Archaea is further classified into Crenarchaea and Euryarchaea. Archaea generally thrive in extreme environments as assessed by temperature, pH, and salinity. For many prokaryotic organisms, ribosomal proteins (RP), transcription/translation factors, and chaperone genes tend to be highly expressed. A gene is predicted highly expressed (PHX) if its codon usage is rather similar to the average codon usage of at least one of the RP, transcription/translation factors, and chaperone gene classes and deviates strongly from the average gene of the genome. The thermosome (Ths) chaperonin family represents the most salient PHX genes among Archaea. The chaperones Trigger factor and HSP70 have overlapping functions in the folding process, but both of these proteins are lacking in most archaea where they may be substituted by the chaperone prefoldin. Other distinctive PHX proteins of Archaea, absent from Bacteria, include the proliferating cell nuclear antigen PCNA, a replication auxiliary factor responsible for tethering the catalytic unit of DNA polymerase to DNA during high-speed replication, and the acidic RP P0, which helps to initiate mRNA translation at the ribosome. Other PHX genes feature Cell division control protein 48 (Cdc48), whereas the bacterial septation proteins FtsZ and minD are lacking in Crenarchaea. RadA is a major DNA repair and recombination protein of Archaea. Archaeal genomes feature a strong Shine Dalgarno ribosome-binding motif more pronounced in Euryarchaea compared with Crenarchaea. PMID- 15883369 TI - Clathrin heavy and light chain isoforms originated by independent mechanisms of gene duplication during chordate evolution. AB - In humans, there are two isoforms each of clathrin heavy chain (CHC17 and CHC22) and light chain (LCa and LCb) subunits, all encoded by separate genes. CHC17 forms the ubiquitous clathrin-coated vesicles that mediate membrane traffic. CHC22 is implicated in specialized membrane organization in skeletal muscle. CHC17 is bound and regulated by LCa and LCb, whereas CHC22 does not functionally interact with either light chain. The imbalanced interactions between clathrin subunit isoforms suggest a distinct evolutionary history for each isoform pair. Phylogenetic and sequence analysis placed both heavy and light chain gene duplications during chordate evolution, 510-600 million years ago. Genes encoding CHC22 orthologues were found in several vertebrate species, with only a pseudogene present in mice. Multiple paralogons surrounding the CHC genes (CLTC and CLTD) were identified, evidence that genomic or large-scale gene duplication produced the two CHC isoforms. In contrast, clathrin light chain genes (CLTA and CLTB) apparently arose by localized duplication, within 1-11 million years of CHC gene duplication. Analysis of sequence divergence patterns suggested that structural features of the CHCs were maintained after gene duplication, but new interactions with regulatory proteins evolved for the CHC22 isoform. Thus, independent mechanisms of gene duplication expanded clathrin functions, concomitant with development of neuromuscular sophistication in chordates. PMID- 15883370 TI - Utility of siRNA against Keap1 as a strategy to stimulate a cancer chemopreventive phenotype. AB - A duplex 21 nucleotide small interfering RNA (siRNA) against human Keap1 is described that represents a unique class of cancer chemopreventive agent. This siRNA can knockdown Keap1 mRNA and thereby relieve negative regulation of nuclear factor erythroid 2 p45-related factor 2 (Nrf2)-mediated gene expression. The siRNA lowered endogenous Keap1 mRNA to <30% of control levels between 24 and 72 h after transfection in human HaCaT keratinocyte cells and was capable of blocking ectopic expression of FLAG-tagged human Keap1 protein but not that of ectopic V5 tagged mouse Keap1 protein. Transfection of human HaCaT cells with Keap1 siRNA markedly enhanced endogenous levels of nuclear factor erythroid 2 p45-related factor 2 (Nrf2) protein and increased transcription of an antioxidant response element-driven reporter gene by 2.3-fold. Furthermore, 48 h after transfection of these cells with Keap1 siRNA, expression of aldo-keto reductase 1C1/2 and the glutamate cysteine ligase catalytic and modifier subunits was elevated between 5- and 14-fold. A modest increase of 3-fold in NAD(P)H:quinone oxidoreductase 1 was also observed. The Keap1 siRNA produced a 1.75-fold increase in intracellular glutathione 48 h after transfection. Thus, antagonism of Keap1 by siRNA can be used to preadapt human cells to oxidative stress without the need to expose them to redox stressors. PMID- 15883371 TI - Patellamide A and C biosynthesis by a microcin-like pathway in Prochloron didemni, the cyanobacterial symbiont of Lissoclinum patella. AB - Prochloron spp. are obligate cyanobacterial symbionts of many didemnid family ascidians. It has been proposed that the cyclic peptides of the patellamide class found in didemnid extracts are synthesized by Prochloron spp., but studies in which host and symbiont cells are separated and chemically analyzed to identify the biosynthetic source have yielded inconclusive results. As part of the Prochloron didemni sequencing project, we identified patellamide biosynthetic genes and confirmed their function by heterologous expression of the whole pathway in Escherichia coli. The primary sequence of patellamides A and C is encoded on a single ORF that resembles a precursor peptide. We propose that this prepatellamide is heterocyclized to form thiazole and oxazoline rings, and the peptide is cleaved to yield the two cyclic patellamides, A and C. This work represents the full sequencing and functional expression of a marine natural product pathway from an obligate symbiont. In addition, a related cluster was identified in Trichodesmium erythraeum IMS101, an important bloom-forming cyanobacterium. PMID- 15883372 TI - Detecting remotely related proteins by their interactions and sequence similarity. AB - The function of an uncharacterized protein is usually inferred either from its homology to, or its interactions with, characterized proteins. Here, we use both sequence similarity and protein interactions to identify relationships between remotely related protein sequences. We rely on the fact that homologous sequences share similar interactions, and, therefore, the set of interacting partners of the partners of a given protein is enriched by its homologs. The approach was bench-marked by assigning the fold and functional family to test sequences of known structure. Specifically, we relied on 1,434 proteins with known folds, as defined in the Structural Classification of Proteins (SCOP) database, and with known interacting partners, as defined in the Database of Interacting Proteins (DIP). For this subset, the specificity of fold assignment was increased from 54% for position-specific iterative BLAST to 75% for our approach, with a concomitant increase in sensitivity for a few percentage points. Similarly, the specificity of family assignment at the e-value threshold of 10(-8) was increased from 70% to 87%. The proposed method would be a useful tool for large-scale automated discovery of remote relationships between protein sequences, given its unique reliance on sequence similarity and protein-protein interactions. PMID- 15883373 TI - Insights into TOR function and rapamycin response: chemical genomic profiling by using a high-density cell array method. AB - With the advent of complete genome sequences, large-scale functional analyses are generating new excitement in biology and medicine. To facilitate genomewide functional analyses, we developed a high-density cell array with quantitative and automated readout of cell fitness. Able to print at > x 10 higher density on a standard microtiter plate area than currently possible, our cell array allows single-plate screening of the complete set of Saccharomyces cerevisiae gene deletion library and significantly reduces the amount of small molecules and other materials needed for the study. We used this method to map the relation between genes and cell fitness in response to rapamycin, a medically important natural product that targets the eukaryotic kinase Tor. We discuss the implications for pharmacogenomics and the uncharted complexity in genotype dependent drug response in molecularly targeted therapies. Our analysis leads to several basic findings, including a class of gene deletions that confer better fitness in the presence of rapamycin. This result provides insights into possible therapeutic uses of rapamycin/CCI-779 in the treatment of neurodegenerative diseases (including Alzheimer's, Parkinson's, and Huntington's diseases), and cautions the possible existence of similar rapamycin-enhanceable mutations in cancer. It is well established in yeast that although TOR2 has a unique rapamycin insensitive function, TOR1 and TOR2 are interchangeable in the rapamycin sensitive functions. We show that even the rapamycin-sensitive functions are distinct between TOR1 and TOR2 and map the functional difference to a approximately 120-aa region at the N termini of the proteins. Finally, we discuss using cell-based genomic pattern recognition in designing electronic or optical biosensors. PMID- 15883374 TI - Dissecting human cytomegalovirus gene function and capsid maturation by ribozyme targeting and electron cryomicroscopy. AB - Human CMV (HCMV) is the leading viral cause of birth defects and causes one of the most common opportunistic infections among transplant recipients and AIDS patients. Cleavage of internal scaffolding proteins by the viral protease (Pr) occurs during HCMV capsid assembly. To gain insight into the mechanism of HCMV capsid maturation and the roles of the Pr in viral replication, an RNase P ribozyme was engineered to target the Pr mRNA and down-regulate its expression by >99%, generating premature Pr-minus capsids. Furthermore, scaffolding protein processing and DNA encapsidation were inhibited by 99%, and viral growth was reduced by 10,000-fold. 3D structural comparison of the Pr-minus and wild-type B capsids by electron cryomicroscopy, at an unprecedented 12.5-angstroms resolution, unexpectedly revealed that the structures are identical in their overall shape and organization. However, the Pr-minus capsid contains tenuous connections between the scaffold and the capsid shell, whereas the wild-type B capsid has extra densities in its core that may represent the viral Pr. Our findings indicate that cleavage of the scaffolding protein is not associated with the morphological changes that occur during capsid maturation. Instead, the protease appears to be required for DNA encapsidation and the subsequent maturation steps leading to infectious progeny. These results therefore provide key insights into an essential step of HCMV infection using an RNase P ribozyme based inhibition strategy. PMID- 15883375 TI - De novo cis-regulatory module elicitation for eukaryotic genomes. AB - Transcription regulation is controlled by coordinated binding of one or more transcription factors in the promoter regions of genes. In many species, especially higher eukaryotes, transcription factor binding sites tend to occur as homotypic or heterotypic clusters, also known as cis-regulatory modules. The number of sites and distances between the sites, however, vary greatly in a module. We propose a statistical model to describe the underlying cluster structure as well as individual motif conservation and develop a Monte Carlo motif screening strategy for predicting novel regulatory modules in upstream sequences of coregulated genes. We demonstrate the power of the method with examples ranging from bacterial to insect and human genomes. PMID- 15883376 TI - Cellulose utilization by Clostridium thermocellum: bioenergetics and hydrolysis product assimilation. AB - The bioenergetics of cellulose utilization by Clostridium thermocellum was investigated. Cell yield and maintenance parameters, Y(X/ATP)True = 16.44 g cell/mol ATP and m = 3.27 mmol ATP/g cell per hour, were obtained from cellobiose grown chemostats, and it was shown that one ATP is required per glucan transported. Experimentally determined values for G(ATP)P-T (ATP from phosphorolytic beta-glucan cleavage minus ATP for substrate transport, mol ATP/mol hexose) from chemostats fed beta-glucans with degree of polymerization (DP) 2-6 agreed well with the predicted value of (n-2)/n [corrected] (n = mean cellodextrin DP assimilated). A mean G(ATP)(P-T) value of 0.52 +/- 0.06 was calculated for cellulose-grown chemostat cultures, corresponding to n = 4.20 +/- 0.46. Determination of intracellular beta-glucan radioactivity resulting from 14C labeled substrates showed that uptake is different for cellulose and cellobiose (G2). For 14C-cellobiose, radioactivity was greatest for G2; substantially smaller but measurable for G1, G3, and G4; undetectable for G5 and G6; and n was approximately 2. For 14C-cellulose, radioactivity was greatest for G5; lower but substantial for G6, G2, and G1; very low for G3 and G4; and n was approximately 4. These results indicate that: (i) C. thermocellum hydrolyzes cellulose by a different mode of action from the classical mechanism involving solubilization by cellobiohydrolase; (ii) bioenergetic benefits specific to growth on cellulose are realized, resulting from the efficiency of oligosaccharide uptake combined with intracellular phosphorolytic cleavage of beta-glucosidic bonds; and (iii) these benefits exceed the bioenergetic cost of cellulase synthesis, supporting the feasibility of anaerobic biotechnological processing of cellulosic biomass without added saccharolytic enzymes. PMID- 15883377 TI - Structural properties of Abeta protofibrils stabilized by a small molecule. AB - Metastable oligomeric and protofibrillar forms of amyloidogenic proteins have been implicated as on-pathway assembly intermediates in amyloid formation and as the major toxic species in a number of amyloid diseases including Alzheimer's disease. We describe here a chemical biology approach to structural analysis of Abeta protofibrils. Library screening yielded several molecules that stimulate Abeta aggregation. One of these compounds, calmidazolium chloride (CLC), rapidly and efficiently converts Abeta(1-40) monomers into clusters of protofibrils. As monitored by electron microscopy, these protofibrils persist for days when incubated in PBS at 37 degrees C, with a slow transition to fibrillar structures apparent only after several weeks. Like normal protofibrils, the CLC-Abeta aggregates exhibit a low thioflavin T response. Like Abeta fibrils, the clustered protofibrils bind the anti-amyloid Ab WO1. The CLC-Abeta aggregates exhibit the same protection from hydrogen-deuterium exchange as do protofibrils isolated from a spontaneous Abeta fibril formation reaction: approximately 12 of the 39 Abeta(1 40) backbone amide protons are protected from exchange in the protofibril, compared with approximately twice that number in amyloid fibrils. Scanning proline mutagenesis analysis shows that the Abeta molecule in these protofibrillar assemblies exhibits the same flexible N and C termini as do mature amyloid fibrils. The major difference in Abeta conformation between fibrils and protofibrils is added structural definition in the 22-29 segment in the fibril. Besides aiding structural analysis, compounds capable of facilitating oligomer and protofibril formation might have therapeutic potential, if they act to sequester Abeta in a form and/or location that cannot engage the toxic pathway. PMID- 15883378 TI - Alternative Gnas gene products have opposite effects on glucose and lipid metabolism. AB - Gnas is an imprinted gene with multiple gene products resulting from alternative splicing of different first exons onto a common exon 2. These products include stimulatory G protein alpha-subunit (G(s)alpha), the G protein required for receptor-stimulated cAMP production; extralarge G(s)alpha (XLalphas), a paternally expressed G(s)alpha isoform; and neuroendocrine-specific protein (NESP55), a maternally expressed chromogranin-like protein. G(s)alpha undergoes tissue-specific imprinting, being expressed primarily from the maternal allele in certain tissues. Heterozygous mutation of exon 2 on the maternal (E2m-/+) or paternal (E2+/p-) allele results in opposite effects on energy metabolism. E2m-/+ mice are obese and hypometabolic, whereas E2+/p- mice are lean and hypermetabolic. We now studied the effects of G(s)alpha deficiency without disrupting other Gnas gene products by deleting G(s)alpha exon 1 (E1). E1+/p- mice lacked the E2+/p- phenotype and developed obesity and insulin resistance. The lean, hypermetabolic, and insulin-sensitive E2+/p- phenotype appears to result from XLalphas deficiency, whereas loss of paternal-specific G(s)alpha expression in E1+/p- mice leads to an opposite metabolic phenotype. Thus, alternative Gnas gene products have opposing effects on glucose and lipid metabolism. Like E2m-/+ mice, E1m-/+ mice had s.c. edema at birth, presumably due to loss of maternal G(s)alpha expression. However, E1m-/+ mice differed from E2m /+ mice in other respects, raising the possibility for the presence of other maternal-specific gene products. E1m-/+ mice had more severe obesity and insulin resistance and lower metabolic rate relative to E1+/p- mice. Differences between E1m-/+ and E1+/p- mice presumably result from differential effects on G(s)alpha expression in tissues where G(s)alpha is normally imprinted. PMID- 15883379 TI - Brain response to putative pheromones in homosexual men. AB - The testosterone derivative 4,16-androstadien-3-one (AND) and the estrogen-like steroid estra-1,3,5(10),16-tetraen-3-ol (EST) are candidate compounds for human pheromones. AND is detected primarily in male sweat, whereas EST has been found in female urine. In a previous positron emission tomography study, we found that smelling AND and EST activated regions covering sexually dimorphic nuclei of the anterior hypothalamus, and that this activation was differentiated with respect to sex and compound. In the present study, the pattern of activation induced by AND and EST was compared among homosexual men, heterosexual men, and heterosexual women. In contrast to heterosexual men, and in congruence with heterosexual women, homosexual men displayed hypothalamic activation in response to AND. Maximal activation was observed in the medial preoptic area/anterior hypothalamus, which, according to animal studies, is highly involved in sexual behavior. As opposed to putative pheromones, common odors were processed similarly in all three groups of subjects and engaged only the olfactory brain (amygdala, piriform, orbitofrontal, and insular cortex). These findings show that our brain reacts differently to the two putative pheromones compared with common odors, and suggest a link between sexual orientation and hypothalamic neuronal processes. PMID- 15883380 TI - Four-dimensional ultrafast electron microscopy. AB - Electron microscopy is arguably the most powerful tool for spatial imaging of structures. As such, 2D and 3D microscopies provide static structures with subnanometer and increasingly with angstrom-scale spatial resolution. Here we report the development of 4D ultrafast electron microscopy, whose capability imparts another dimension to imaging in general and to dynamics in particular. We demonstrate its versatility by recording images and diffraction patterns of crystalline and amorphous materials and images of biological cells. The electron packets, which were generated with femtosecond laser pulses, have a de Broglie wavelength of 0.0335 angstroms at 120 keV and have as low as one electron per pulse. With such few particles, doses of few electrons per square angstrom, and ultrafast temporal duration, the long sought after but hitherto unrealized quest for ultrafast electron microscopy has been realized. Ultrafast electron microscopy should have an impact on all areas of microscopy, including biological imaging. PMID- 15883381 TI - The neurotrophin receptor p75NTR modulates long-term depression and regulates the expression of AMPA receptor subunits in the hippocampus. AB - Neurotrophins are involved in the modulation of synaptic transmission, including the induction of long-term potentiation (LTP) through the receptor TrkB. Because previous studies have revealed a bidirectional mode of neurotrophin action by virtue of signaling through either the neurotrophin receptor p75NTR or the Trk receptors, we tested the hypothesis that p75NTR is important for longterm depression (LTD) to occur. Although LTP was found to be unaffected in hippocampal slices of two different strains of mice carrying mutations of the p75NTR gene, hippocampal LTD was impaired in both p75NTR-deficient mouse strains. Furthermore, the expression levels of two (RS)-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor subunits, GluR2 and GluR3, but not GluR1 or GluR4, were found to be significantly altered in the hippocampus of p75NTR deficient mice. These results implicate p75NTR in activity-dependent synaptic plasticity and extend the concept of functional antagonism of the neurotrophin signaling system. PMID- 15883382 TI - Serum IgG mediates mucosal immunity against rotavirus infection. AB - We evaluated the protective role of passively transferred circulating antibodies in protecting non-human primates against experimental rotavirus infection. Pooled sera with rotavirus-specific IgG titers that were either high (1:10,000), intermediate (1:300), or negative (< 1:25) were infused i.v. into naive pigtailed macaques (ages 3-6 months). Rotavirus-specific IgG could be detected in the sera at 18 h in all animals infused with antibody-containing serum, and fecal IgG titers could be detected only in animals given high-titer pooled sera. When orally challenged with 10(6) fluorescent-forming units of a simian rotavirus strain, YK-1, at 18 h after serum transfer, control animals shed virus starting 1 3 days after challenge and continued to shed virus at high titers for 6-8 days, whereas passively immunized macaques did not shed virus or had delayed shedding at low titers for only a limited time. The observation that passively transferred antibodies can suppress or delay viral infection in rotavirus-challenged pigtailed macaques has important implications for the design and testing of parenteral candidate rotavirus vaccines. PMID- 15883383 TI - The Nkx6.1 homeodomain transcription factor suppresses glucagon expression and regulates glucose-stimulated insulin secretion in islet beta cells. AB - We have previously described rat insulinoma INS-1-derived cell lines with robust or poor glucose-stimulated insulin secretion (GSIS). In the current study, we have further resolved these lines into three classes: class 1, glucose unresponsive/glucagon-expressing; class 2, glucose-unresponsive/glucagon negative; and class 3, glucose-responsive/glucagon-negative. The transcription factor Nkx2.2 was expressed with relative abundance of 3.3, 1.0, and 1.0 in class 1, class 2, and class 3 cells, respectively, whereas Nkx6.1 expression had the opposite trend: 1.0, 2.6, and 6.4 in class 1, class 2, and class 3 cells, respectively. In class 1 cells, overexpressed Nkx6.1 suppressed glucagon expression but did not affect the levels of several other prominent beta cell transcription factors. RNA interference (RNAi)-mediated suppression of Nkx6.1 in class 3 cells resulted in a doubling of glucagon mRNA, with no effect on Pdx1 levels, whereas suppression of Pdx1 in class 3 cells caused a 12-fold increase in glucagon transcript levels, demonstrating independent effects of Nkx6.1 and Pdx1 on glucagon expression in beta cell lines. RNAi-mediated suppression of Nkx6.1 expression in class 3 cells also caused a decrease in GSIS from 13.9- to 3.7 fold, whereas suppression of Pdx1 reduced absolute amounts of insulin secretion without affecting fold response. Finally, RNAi-mediated suppression of Nkx6.1 mRNA in primary rat islets was accompanied by a significant decrease in GSIS relative to control cells. In sum, our studies have revealed roles for Nkx6.1 in suppression of glucagon expression and control of GSIS in islet beta cells. PMID- 15883384 TI - Functional organization of human occipital-callosal fiber tracts. AB - Diffusion tensor imaging (DTI) and fiber tracking (FT) were used to measure the occipital lobe fiber tracts connecting the two hemispheres in individual human subjects. These tracts are important for normal vision. Also, damage to portions of these tracts is associated with alexia. To assess the reliability of the DTI FT measurements, occipital-callosal projections were estimated from each subject's left and right hemispheres independently. The left and right estimates converged onto the same positions within the splenium. We further characterized the properties of the estimated occipital-callosal fiber tracts by combining them with functional MRI. We used functional MRI to identify visual field maps in cortex and labeled fibers by the cortical functional response at the fiber endpoint. This labeling reveals a regular organization of the fibers within the splenium. The dorsal visual maps (dorsal V3, V3A, V3B, V7) send projections through a large band in the middle of the splenium, whereas ventral visual maps (ventral V3, V4) send projections through the inferior-anterior corner of the splenium. The agreement between the independent left/right estimates, further supported by previous descriptions of homologous tracts in macaque, validates the DTI-FT methods. However, a principal limitation of these methods is low sensitivity: a large number of fiber tracts that connect homotopic regions of ventral and lateral visual cortex were undetected. We conclude that most of the estimated tracts are real and can be localized with a precision of 1-2 mm, but many tracts are missed because of data and algorithm limitations. PMID- 15883385 TI - The use of oscillatory signals in the study of genetic networks. AB - The structure of a genetic network is uncovered by studying its response to external stimuli (input signals). We present a theory of propagation of an input signal through a linear stochastic genetic network. We found that there are important advantages in using oscillatory signals over step or impulse signals and that the system may enter into a pure fluctuation resonance for a specific input frequency. PMID- 15883386 TI - Coevolution of TCR-MHC interactions: conserved MHC tertiary structure is not sufficient for interactions with the TCR. AB - The specificity for self-MHC that is necessary for T cell function is a consequence of intrathymic selection during which T cell antigen receptors (TCRs) expressed by immature thymocytes are tested for their affinity for self peptide:self-MHC. The germ-line-encoded segments of the TCR, however, are believed to have an innate specificity for structural features of MHC molecules. We directly tested this hypothesis by generating a transgenic mouse system in which the protein HLA-DM is expressed at the surface of thymic cortical epithelial cells in the absence of classical MHC molecules. The specialized intracellular function of HLA-DM has removed this MHC class II-like protein from the evolutionary forces that have been hypothesized to shape TCR-MHC interactions. Our study shows that a structural mimic of MHC class II is not sufficient to appropriately interact with the TCRs expressed by developing thymocytes. This result emphasizes the unique complementarity of TCR-MHC interactions that are maintained by the evolutionary pressures dictated by positive selection. PMID- 15883387 TI - Role of peripheral corticotropin-releasing factor and urocortin II in intestinal inflammation and motility in terminal ileum. AB - Corticotropin-releasing factor (CRF) and the closely related family of neuropeptides urocortins (Ucns) are ancient paracrine-signaling peptides secreted in both the central and peripheral neural circuits. CRF and Ucns released from the CNS (central) regulate a plethora of physiological processes that include food intake, inflammation, and bowel motility and permeability. In the gastrointestinal tract, CRF actions are largely proinflammatory, whereas the effects of the Ucn subtypes can be either pro- or antiinflammatory. Central (intracerebroventricular) or peripheral (i.p.) administration of CRF or Ucns inhibits gastric emptying and promotes colonic motility. To ascertain the role of peripherally expressed CRF and UcnII in gastrointestinal inflammation and motility, we generated ileum-specific phenotypic knockouts of these peptides by using RNA interference. Long dsRNA effectively silenced basal expression of CRF and UcnII in ileum. Control dsRNA or saline treatment did not affect CRF or UcnII expression. In an experimental model of toxin-induced intestinal inflammation, inhibition of CRF ablated the inflammatory response (measured by epithelial damage, mucosal edema, and neutrophil infiltration). UcnII dsRNA treatment did not alter the inflammatory response to toxin. Furthermore, ileal motility was increased after site-specific inhibition of both CRF and UcnII. Thus, we demonstrate that ileal-specific CRF promotes inflammation and both CRF and UcnII modulate bowel motility. PMID- 15883388 TI - Investigating local conformations of double-stranded DNA by low-energy circular dichroism of pyrrolo-cytosine. AB - Local base stacking and conformational mobility play a major role in the structure and function of nucleic acids. We have recently shown that the low energy CD spectrum of 2-aminopurine (2-AP), i.e., the CD spectral region above 300 nm, can be used to monitor conformational changes in polynucleotides at or near mono- and dinucleotide 2-AP residues that replace adenine residues in DNA and RNA. Here, we extend this technique to pyrrolo-cytosine (PC), a fluorescent analogue of cytosine. The low-energy CD spectrum of a PC dinucleotide in dsDNA exhibits an exciton couplet with two bands of opposite sign centered at 350 nm. This signal is characteristic of base stacking between adjacent PC residues in a right helical conformation. Isolated PC nucleotide residues inserted into polynucleotide chains also display chirality that reflects the asymmetric environment of their sequence context. Thus, we show that the low-energy CD spectra of C(PC)A and A(PC)C sequences in dsDNA have opposite signs. It appears that the measurement of the low-energy CD spectra of PC residues will usefully complement 2-AP measurements by serving to characterize the local conformations and dynamics of nucleic acids near C residues and G-C base pairs. PMID- 15883389 TI - Prescription drug expenditures rising--too fast? PMID- 15883390 TI - Lawnmower neuritis: an unusual occupational hazard. PMID- 15883391 TI - Body weight classification. PMID- 15883393 TI - Wait Time Alliance first to set benchmarks. PMID- 15883394 TI - Universite de Montreal in the dark about fraud. PMID- 15883395 TI - Quebec medical students strike at their own pace. PMID- 15883396 TI - Canadians spending more on drugs. PMID- 15883397 TI - C. difficile strain 20 times more virulent. PMID- 15883398 TI - A national strategy for waiting-times research? PMID- 15883399 TI - How an imprint can lead to cancer. PMID- 15883400 TI - Vaginal bleeding in the prepubertal child. PMID- 15883401 TI - Preventing violence in youth. PMID- 15883402 TI - Does intensified cholesterol lowering provide greater protection from cardiovascular events among patients with stable coronary artery disease? PMID- 15883403 TI - Intracranial hematoma in a patient with AIDS. PMID- 15883404 TI - Insulin resistance syndrome, body mass index and the risk of ischemic heart disease. AB - BACKGROUND: Many people who are not obese according to standard height and weight criteria may still display features of insulin resistance syndrome and thus be at high risk of ischemic heart disease. We sought to investigate the effect of cumulative features of insulin resistance syndrome on the risk of ischemic heart disease associated with variations in body mass index (BMI) among men who participated in the Quebec Cardiovascular Study. METHODS: A cohort of 1824 nondiabetic men free of ischemic heart disease was evaluated at the 1985 baseline evaluation and followed for a period of 13 years, during which 284 first ischemic heart disease events were recorded. Relative hazards (RHs) of ischemic heart disease in 3 BMI groups (normal weight, overweight and obese) were estimated using Cox proportional hazards regression. RESULTS: Although obese men (BMI > or = 30 kg/m2) were the most likely to accumulate features of insulin resistance syndrome, the univariate risk of ischemic heart disease in this group was not significantly increased compared with normal-weight men (BMI < 25 kg/m2) (RH 1.26, 95% confidence interval [CI] 0.88-1.80). However, obese men who accumulated more than 4 features of insulin resistance syndrome were at increased risk of ischemic heart disease (RH 1.81, 95% CI 1.02-3.19) compared with normal-weight men who had fewer than 3 features of the syndrome. Conversely, having more than 4 features of insulin resistance syndrome was associated with a 3-fold increase in the risk of ischemic heart disease among normal-weight men (RH 3.01, 95% CI 1.70 5.32). INTERPRETATION: Although obesity is an important risk factor for ischemic heart disease, variations in BMI alone poorly reflect the risk of ischemic heart disease associated with features of insulin resistance syndrome. PMID- 15883405 TI - Thrombolysis for acute ischemic stroke: results of the Canadian Alteplase for Stroke Effectiveness Study. AB - BACKGROUND: Thrombolysis for acute ischemic stroke has remained controversial. The Canadian Alteplase for Stroke Effectiveness Study, a national prospective cohort study, was conducted to assess the effectiveness of alteplase therapy for ischemic stroke in actual practice. METHODS: The study was mandated by the federal government as a condition of licensure of alteplase for the treatment of stroke in Canada. A registry was established to collect data over 2.5 years for stroke patients receiving such treatment from Feb. 17, 1999, through June 30, 2001. All centres capable of administering thrombolysis therapy according to Canadian guidelines were eligible to submit patient data to the registry. Data collection was prospective, and follow-up was completed at 90 days after stroke. Copies of head CT scans obtained at baseline and at 24-48 hours after the start of treatment were submitted to a central panel for review. RESULTS: A total of 1135 patients were enrolled at 60 centres in all major hospitals across Canada. The registry collected data for an estimated 84% of all treated ischemic stroke patients in the country. An excellent clinical outcome was observed in 37% of the patients. Symptomatic intracranial hemorrhage occurred in only 4.6% of the patients (95% confidence interval [CI] 3.4%-6.0%); however, 75% of these patients died in hospital. An additional 1.3% (95% CI 0.7%-2.2%) of patients had hemiorolingual angioedema. CONCLUSIONS: The outcomes of stroke patients undergoing thrombolysis in Canada are commensurate with the results of clinical trials. The rate of symptomatic intracranial hemorrhage was low. Stroke thrombolysis is a safe and effective therapy in actual practice. PMID- 15883406 TI - Heart disease risk among metabolically healthy obese men and metabolically unhealthy lean men. PMID- 15883407 TI - Clinical practice guidelines for the care and treatment of breast cancer: follow up after treatment for breast cancer (summary of the 2005 update). PMID- 15883408 TI - Canadian prescription drug costs surpass 18 billion dollars. PMID- 15883409 TI - Women's sexual dysfunction: revised and expanded definitions. AB - Acceptance of an evidence-based conceptualization of women's sexual response combining interpersonal, contextual, personal psychological and biological factors has led to recently published recommendations for revision of definitions of women's sexual disorders found in the American Psychiatric Association's Diagnostic and Statistical Manual (DSM-IV-TR). DSM-IV definitions have focused on absence of sexual fantasies and sexual desire prior to sexual activity and arousal, even though the frequency of this type of desire is known to vary greatly among women without sexual complaints. DSM-IV definitions also focus on genital swelling and lubrication, entities known to correlate poorly with subjective sexual arousal and pleasure. The revised definitions consider the many reasons women agree to or instigate sexual activity, and reflect the importance of subjective sexual arousal. The underlying conceptualization of a circular sex response cycle of overlapping phases in a variable order may facilitate not only the assessment but also the management of dysfunction, the principles of which are briefly recounted. PMID- 15883410 TI - Phase II study of low-dose decitabine in patients with chronic myelogenous leukemia resistant to imatinib mesylate. AB - PURPOSE: To determine the activity of decitabine, a DNA methylation inhibitor, in imatinib-refractory or intolerant chronic myelogenous leukemia. MATERIALS AND METHODS: Thirty-five patients were enrolled in this phase II study (12 in chronic phase, 17 in accelerated phase, and six in blastic phase). Decitabine was administered at 15 mg/m2 intravenously over 1 hour daily, 5 days a week for 2 weeks. DNA methylation was measured using a LINE1 bisulfite/pyrosequencing assay. RESULTS: Complete hematologic responses were seen in 12 patients (34%) and partial hematologic responses in seven patients (20%), for an overall hematologic response rate of 54% (83% in chronic phase, 41% in accelerated phase, and 34% in blastic phase). Major cytogenetic responses were observed in six patients (17%), and minor cytogenetic responses were seen in 10 patients (29%) for an overall cytogenetic response rate of 46%. Median response duration was 3.5 months (range, 2 to 13+ months). Myelosuppression was the major adverse effect, with neutropenic fever in 28 (23%) of 124 courses of therapy. LINE1 methylation decreased from 71.3% +/- 1.4% (mean +/- standard error of the mean) to 60.7% +/- 1.4% after 1 week, 50.9% +/- 2.4% after 2 weeks, and returned to 66.5% +/- 2.7% at recovery of counts (median, 46 days). LINE1 methylation at the end of week 1 did not correlate with subsequent responses. However, at day 12, the absolute decrease in methylation was 14.5% +/- 3.0% versus 26.8% +/- 2.7% in responders versus nonresponders (P = .007). CONCLUSION: Decitabine induces hypomethylation and has clinical activity in imatinib refractory chronic myelogenous leukemia. We hypothesize that the inverse correlation between hypomethylation 2 weeks after therapy and response is due to a cell death mechanism of response, whereby resistant cells can withstand more hypomethylation. PMID- 15883411 TI - AIDS-related Burkitt's lymphoma versus diffuse large-cell lymphoma in the pre highly active antiretroviral therapy (HAART) and HAART eras: significant differences in survival with standard chemotherapy. AB - PURPOSE: To compare outcomes of patients with HIV-Burkitt's lymphoma (HIV-BL) and HIV-diffuse large-cell lymphoma (HIV-DLCL) after treatment with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) or M-BACOD (methotrexate, bleomycin, cyclophosphamide, etoposide) in pre-highly active antiretroviral therapy (HAART) versus HAART eras. PATIENTS AND METHODS: Three hundred sixty-three patients with AIDS-related lymphoma diagnosed from 1982 to 2003 were reviewed retrospectively, including 262 in the pre-HAART (HIV-BL, 117; HIV-DLCL, 145) and 101 in the HAART era (HIV-BL, 18; HIV-DLCL, 83). Pre-HAART included those who did not receive HAART, and HAART era included those diagnosed after January 1997 who received HAART. RESULTS: There were no significant differences between groups in terms of age, sex, history of injection drug use, prior AIDS, lactate dehydrogenase level, and disease stage at diagnosis. Compared with HIV-BL, HIV-DLCL was associated with significantly lower CD4 counts in the pre-HAART but not the HAART era. Although the overall median survival was similar for both groups in the pre-HAART era (HIV-BL, 6.4 months v HIV-DLCL, 8.3 months; P = .43), survival was significantly worse in patients with HIV-BL in the HAART era (HIV-BL, 5.7 months v HIV-DLCL, 43.2 months; P = .0003). Failure to attain complete remission and CD4 count less than 100 cells/mm(3) independently predicted for poor survival in the pre-HAART era. In comparison, histology of HIV BL and no attainment of complete remission were independent poor prognostic factors in the HAART era. CONCLUSION: Survival of patients with HIV-DLCL has improved in the HAART era, along with CD4 count, whereas survival of similarly treated patients with HIV-BL remained poor. The current practice of using the same regimen for both groups of patients should be re-evaluated. PMID- 15883412 TI - Extent of damage and repair in the p53 tumor-suppressor gene after treatment of myeloma patients with high-dose melphalan and autologous blood stem-cell transplantation is individualized and may predict clinical outcome. AB - PURPOSE: To quantitate the individual levels of melphalan-induced DNA damage formation and repair in vivo and to search for possible correlations with clinical outcome in patients with multiple myeloma (MM). PATIENTS AND METHODS: The formation and subsequent repair of DNA damage (monoadducts and interstrand cross-links) in the p53 tumor-suppressor gene, the proto-oncogene N-ras, and the housekeeping gene beta-actin during the first 24 hours after treatment with high dose melphalan (HDM; 200 mg/m2) supported by autologous blood stem-cell transplantation (ABSCT) was measured in blood leukocytes of 26 patients with MM. The peak DNA adduct levels, the total amount of adducts over time, and the rate of adducts repair in each gene were correlated with response and time to progression after HDM. RESULTS: The levels of gene-specific DNA damage formation and the individual repairing capacity varied up to 16-fold among patients, indicating that the melphalan-induced biologic effect in vivo is highly individualized. A significantly greater DNA damage and a slower rate of repair in p53 for all end points under study were found in patients who achieved tumor reduction compared with nonresponding patients. Furthermore, longer progression free survival correlated with increased peak monoadduct levels in the p53 gene (P = .032). CONCLUSION: Increased DNA damage and slower repairing capacity in the p53 gene from blood leukocytes after HDM correlate with improved outcome of patients with MM who undergo ABSCT. These results suggest that quantitation of such biologic end points may identify patients who are more likely to benefit from this procedure. PMID- 15883413 TI - Optimizing the cardiovascular outcomes of weight loss. PMID- 15883414 TI - B vitamins and cognitive function: do we need more and larger trials? PMID- 15883415 TI - Aging muscle. AB - Age causes structural and functional changes in skeletal muscle in a wide range of species, including humans. Muscle changes in humans start in the fourth decade of life and cause frailty and disabilities. Associated changes in body composition form the basis of many metabolic disorders, such as insulin resistance, type 2 diabetes, hypertension, and hyperlipidemia, which result in an increased incidence of cardiovascular death. Decreases in the synthesis rates of many muscle proteins, specifically of myosin heavy chain and mitochondrial proteins, occur with age. The underlying causes of the reduction in mitochondrial biogenesis and ATP production seem to be decreases in mitochondrial DNA and messenger RNA. Reduced ATP production could be the basis of reduced muscle protein turnover, which requires energy. Both aerobic exercise and resistance exercise enhance muscle protein synthesis and mitochondrial biogenesis. Insulin and amino acids have also been shown to enhance muscle mitochondrial biogenesis and mitochondrial protein synthesis. However, the insulin-induced increase in muscle mitochondrial ATP production is defective in type 2 diabetic patients with insulin resistance. Moreover, a dissociation between increases in muscle mitochondrial biogenesis and insulin sensitivity after exercise has been noted in older persons. It remains to be determined whether muscle mitochondrial dysfunction causes or results from insulin resistance. Exercise seems to enhance the efficiency of muscle mitochondrial DNA in rodents. Reduced physical activity as a contributor of age-related mitochondrial dysfunction remains to be determined. It is proposed that a reduction in tissue mitochondrial ATP production signals the hypothalamic centers to reduce spontaneous physical activities. Voluntary physical activity is regulated by cognitive centers and could attenuate the progressive decline in mitochondrial functions that occurs with age. PMID- 15883416 TI - Physical activity energy expenditure predicts changes in body composition in middle-aged healthy whites: effect modification by age. AB - BACKGROUND: It is unclear whether physical activity energy expenditure (PAEE) predicts changes in body composition. OBJECTIVE: The objective was to describe the independent associations between PAEE and changes in body composition in a population-based cohort. DESIGN: This was a prospective population-based study conducted in 739 (311 men and 428 women) healthy middle-aged (median age: 53.8 y) whites. The median follow-up was 5.6 y. PAEE (MJ/d) was assessed by heart rate monitoring, individually calibrated by using the FLEX heart rate method. Fat mass (FM) and fat-free mass (FFM) were assessed by bioimpedance. RESULTS: Body weight (BW) at follow-up was significantly related to baseline PAEE (P < 0.05) after adjustment for sex, baseline age, FM, FFM, and follow-up time. A significant interaction between PAEE and age (P = 0.023) was observed. After the subjects were stratified (above and below the median for age), BW increased by a mean (+/ SD) of 1.7 +/- 5.9 kg (P < 0.0001) in the younger cohort. In this group, follow up FM was significantly associated with baseline PAEE (P = 0.036) after adjustment for confounders. In the older cohort, BW did not change between baseline and follow-up. In this group, in contrast with the younger population, follow-up BW, FM, and FFM were all significantly and positively associated with baseline PAEE (P < 0.01 for all). CONCLUSIONS: Baseline PAEE predicts a change in FM in younger adults, who as a group gained weight in this study. In contrast, baseline PAEE in older adults--who were on average weight stable--is associated with a gain in BW, which was explained by an increase in FM and FFM. PMID- 15883417 TI - Obesity and risk factors for the metabolic syndrome among low-income, urban, African American schoolchildren: the rule rather than the exception? AB - BACKGROUND: Adult obesity is associated with the metabolic syndrome; however, the prevalence of the metabolic syndrome among young children has not been reported. Clinic-based screening efforts for the metabolic syndrome in low-income neighborhoods, where obesity is prevalent, are limited by minimal health insurance coverage and inadequate access to health care. School-based obesity screening programs may effectively target high-risk populations. OBJECTIVE: The objective was to describe the prevalence of overweight and features of the metabolic syndrome (defined as the presence of > or =3 of the following risk factors: HDL < or = 40 mg/dL, triacylglycerol > or = 110 mg/dL, and blood pressure or waist circumference at or above the 90th percentile) in a pilot, school-based screening program. DESIGN: A cross-sectional study of obesity and the metabolic syndrome was conducted in third- to sixth-grade, low-income, urban, African American children. Lipid and glucose concentrations were measured in fasting capillary finger-stick samples. RESULTS: Age- and sex-specific BMI percentiles were assessed in 385 students, 90 of whom were full participants in this study (participants) and 295 of whom had only height and weight measurements taken (other students). Risk factors of the metabolic syndrome were assessed in the 90 participants (23%). No significant differences in BMI percentiles were found between the participants and the other students. Overall, 44% of the participants had BMIs at or above the 85th percentile, and 59% had an elevated BMI or one metabolic syndrome risk factor. The metabolic syndrome was present in 5.6% of all participants, in 13.8% of participants with BMIs at or above the 95th percentile, and in 0% of participants with BMIs below the 95th percentile. CONCLUSIONS: Most of the African American children attending 2 urban schools in low-income neighborhoods were overweight or had one or more risk factors for the metabolic syndrome. School-based screening programs in high-risk populations may provide an efficient venue for the screening of obesity and related risk factors. PMID- 15883418 TI - Effects of an ad libitum low-glycemic load diet on cardiovascular disease risk factors in obese young adults. AB - BACKGROUND: The optimal nutritional approach for the prevention of cardiovascular disease among obese persons remains a topic of intense controversy. Available approaches range from conventional low-fat to very-low-carbohydrate diets. OBJECTIVE: The aim of this pilot study was to evaluate the efficacy of an ad libitum low-glycemic load diet, without strict limitation on carbohydrate intake, as an alternative to a conventional low-fat diet. DESIGN: A randomized controlled trial compared 2 dietary treatments in obese young adults (n = 23) over 12 mo. The experimental treatment emphasized ad libitum consumption of low-glycemic index foods, with 45-50% of energy from carbohydrates and 30-35% from fat. The conventional treatment was restricted in energy (250-500 kcal/d deficit) and fat (<30% of energy), with 55-60% of energy from carbohydrate. We compared changes in study outcomes by repeated-measures analysis of log-transformed data and expressed the results as mean percentage change. RESULTS: Body weight decreased significantly over a 6-mo intensive intervention in both the experimental and conventional diet groups (-8.4% and -7.8%, respectively) and remained below baseline at 12 mo (-7.8% and -6.1%, respectively). The experimental diet group showed a significantly greater mean decline in plasma triacylglycerols than did the conventional diet group (-37.2% and -19.1%, respectively; P = 0.005). Mean plasminogen activator inhibitor 1 concentrations decreased (-39.0%) in the experimental diet group but increased (33.1%) in the conventional diet group (P = 0.004). Changes in cholesterol concentrations, blood pressure, and insulin sensitivity did not differ significantly between the groups. CONCLUSION: An ad libitum low-glycemic load diet may be more efficacious than a conventional, energy-restricted, low-fat diet in reducing cardiovascular disease risk. PMID- 15883419 TI - Blood pressure change with weight loss is affected by diet type in men. AB - BACKGROUND: Weight loss reduces blood pressure, and the Dietary Approaches to Stop Hypertension (DASH) diet has also been shown to lower blood pressure. OBJECTIVE: Our goal was to assess the effect on blood pressure of 2 weight reduction diets: a low-fat diet (LF diet) and a moderate-sodium, high-potassium, high-calcium, low-fat DASH diet (WELL diet). DESIGN: After baseline measurements, 63 men were randomly assigned to either the WELL or the LF diet for 12 wk, and both diet groups undertook 0.5 h of moderate physical activity on most days of the week. RESULTS: Fifty-four men completed the study. Their mean (+/-SD) age was 47.9 +/- 9.3 y (WELL diet, n = 27; LF diet, n = 27), and their mean baseline home systolic and diastolic blood pressures were 129.4 +/- 11.3 and 80.6 +/- 8.6 mm Hg, respectively. Body weight decreased by 4.9 +/- 0.6 kg (+/-SEM) in the WELL group and by 4.6 +/- 0.6 kg in the LF group (P < 0.001 for both). There was a greater decrease in blood pressure in the WELL group than in the LF group [between-group difference (week 12 -baseline) in both SBP (5.5 +/- 1.9 mm Hg; P = 0.006) and DBP (4.4 +/- 1.2 mm Hg; P = 0.001)]. CONCLUSIONS: For a comparable 5 kg weight loss, a diet high in low-fat dairy products, vegetables, and fruit (the WELL diet) resulted in a greater decrease in blood pressure than did the LF diet. This dietary approach to achieving weight reduction may confer an additional benefit in reducing blood pressure in those who are overweight. PMID- 15883420 TI - Plasma lycopene, other carotenoids, and retinol and the risk of cardiovascular disease in men. AB - BACKGROUND: Emerging evidence suggests a possible role of lycopene in the primary prevention of cardiovascular disease (CVD). OBJECTIVE: We examined whether plasma lycopene concentrations in the Physicians' Health Study were associated with CVD in a prospective, nested, case-control design. DESIGN: Baseline blood samples were collected starting in 1996. During a mean follow-up of 2.1 y, we identified 499 cases of CVD (confirmed myocardial infarction, stroke, CVD death, or revascularization procedures) and an equal number of men free of CVD and matched for age (x: 69.7 y), follow-up time, and smoking status. We collected self reported coronary disease risk factors and measured plasma carotenoids, retinol, lipids, and C-reactive protein. RESULTS: In matched analyses with additional adjustment for plasma total cholesterol and randomized treatment, the relative risks (RRs) of CVD for men in the lowest to highest quartiles of plasma lycopene were 1.00 (reference), 0.92, 1.04, and 0.95 (P for linear trend = 0.93). With multivariate adjustment, the RRs of total CVD were 1.00 (reference), 1.08, 0.94, and 1.03 (P for linear trend = 0.98). For important vascular events (241 cases), excluding revascularization procedures, the multivariate RRs remained nonsignificant (P for linear trend = 0.50). Adding plasma carotenoids, lipids, or C-reactive protein to multivariate models had a minimal effect on the RRs of total CVD for plasma lycopene. Compared with lycopene, higher concentrations of plasma lutein/zeaxanthin and retinol suggested a moderate increase in CVD risk, whereas no association was found for beta-cryptoxanthin, alpha-carotene, and beta carotene. CONCLUSIONS: Higher plasma lycopene concentrations were not associated with the risk of CVD in this study of older men. Further evaluation in diverse populations is necessary. PMID- 15883421 TI - Dietary antioxidants and fat are associated with plasma antibody titers to heat shock proteins 60, 65, and 70 in subjects with dyslipidemia. AB - BACKGROUND: The heat shock proteins (HSPs) are protein chaperones. Higher titers of antibody to HSPs (anti-HSPs) have been reported in atherosclerosis, which may contribute to immunoactivation in this process. OBJECTIVE: We investigated whether dietary antioxidants and fat intake are associated with changes in anti HSP titers in dyslipidemic subjects. DESIGN: Patients (n = 238) were recruited from hospital lipid clinics. Control subjects (n = 188) were recruited from university and hospital employees. Food-frequency questionnaires were used to estimate dietary antioxidants and fat. RESULTS: Dyslipidemic patients had significantly higher titers of anti-HSPs than did control subjects; expressed in medians and interquartile ranges of absorbance units, anti-HSP-60 titers were 0.27 (0.18-0.37) and 0.22 (0.16-0.30), anti-HSP-65 titers were 0.45 (0.28-0.79) and 0.31 (0.22-0.50), and anti-HSP-70 titers were 0.22 (0.17-0.30) and 0.19 (0.13 0.27), respectively. Median and interquartile ranges of serum concentrations of C reactive protein [1.25 (0.42-3.26) and 0.58 (0.17-1.42)] and mean (+/-SEM) concentrations of vitamin E (16.36 +/- 0.31 and 14.08 +/- 0.38) were also significantly higher in patients than in control subjects, respectively. In dyslipidemic patients, the major dietary predictors of the variability in anti HSP-60 titers were vitamin C (P = 0.005), vitamin E (P = 0.04), and total fat (P = 0.009) intakes; for anti-HSP-65 titers, vitamin C was the major predictor (P = 0.002). These findings remained significant after adjustment for confounding factors. CONCLUSIONS: Anti-HSP-60, -65, and -70 titers are significantly higher in dyslipidemic patients with or without established coronary disease. Our data indicate an association between dietary constituents and the immune response to HSPs in dyslipidemic subjects. PMID- 15883422 TI - TAS2R38 (phenylthiocarbamide) haplotypes, coronary heart disease traits, and eating behavior in the British Women's Heart and Health Study. AB - BACKGROUND: Variation in the perception of bitter tastes has been associated with eating behavior, body composition, and cardiovascular disease. Recent observations have implicated 2 common haplotypes of TAS2R38 in the determination of bitter compound-tasting ability. OBJECTIVE: The objectives of the study were to examine, in the British Women's Heart and Health Study cohort, any association between TAS2R38 haplotypes, coronary heart disease (CHD), CHD risk factors, and eating behavior and to examine whether the associations allow for estimation of the effects of variation in diet on the etiology of common disease. DESIGN: We conducted a cross-sectional study of relations between TAS2R38 haplotypes and CHD, CHD risk factors, and eating behavior in 3383 women from 23 British towns. RESULTS: Genotyping at P49A and V262A in TAS2R38 (rs713598 and rs1726866) allowed construction of all 4 possible haplotypes. The 2 most common haplotypes corresponded with previously identified haplotypes related to bitter compound tasting ability. No substantial evidence of association was found between these haplotypes and CHD (odds ratio for taste-defining haplotypes: 0.97; 95% CI: 0.78, 1.2), body mass index (difference between means of taste-defining haplotypes: 0.084; 95% CI: -0.45, 0.29), or a series of physiologic and dietary characteristics. A marginally lower risk of diabetes was observed among those with the nontaster haplotype than among those with the taster haplotype (odds ratio: 0.69; 95% CI: 0.48, 1.00). CONCLUSION: TAS2R38 status was not an important determinant of CHD, related risk factors, or eating behavior in the British Women's Heart and Health Study sample. PMID- 15883423 TI - Longitudinal study of soy food intake and blood pressure among middle-aged and elderly Chinese women. AB - BACKGROUND: Several small-scale clinical trials have suggested a potential beneficial effect of short-term soy consumption on blood pressure (BP). Data are scanty on long-term effects of the usual intake of soy foods on BP in general populations. OBJECTIVE: Our aim was to examine the association between usual intake of soy foods and BP. DESIGN: The usual intake of soy foods was assessed at baseline, and BP was measured 2-3 y after the baseline survey among 45 694 participants of the Shanghai Women's Health Study aged 40-70 y who had no history of hypertension, diabetes, or cardiovascular disease at recruitment. Multiple regression models were used to estimate mean differences in BP associated with various intakes of soy foods. RESULTS: Soy protein intake was inversely associated with both systolic BP (P for trend = 0.01) and diastolic BP (P for trend = 0.009) after adjustment for age, body mass index, and lifestyle and other dietary factors. The adjusted mean systolic BP was 1.9 mm Hg lower (95% CI: -3.0, -0.8 mm Hg) and the diastolic BP was 0.9 mm Hg lower (-1.6, -0.2 mm Hg) in women who consumed > or =25 g soy protein/d than in women consuming <2.5 g/d. The inverse associations became stronger with increasing age (P for interaction < 0.05 for both BPs). Among women >60 y old, the corresponding differences were 4.9 mm Hg (95% CI: -8.0, -1.9 mm Hg) for systolic BP and -2.2 mm Hg (95% CI: 3.8, -0.6 mm Hg) for diastolic BP. CONCLUSION: Usual intake of soy foods was inversely associated with both systolic and diastolic BPs, particularly among elderly women. PMID- 15883424 TI - QDR 4500A dual-energy X-ray absorptiometer underestimates fat mass in comparison with criterion methods in adults. AB - BACKGROUND: Dual-energy X-ray absorptiometry (DXA) has become one of the most frequently used methods for estimating human body composition. Although the DXA technique has been validated for the measurement of fat-free mass and fat mass, differences in calibration between instruments produced by different manufacturers, as well as between different models produced by the same manufacturer, have been reported. OBJECTIVE: The objective was to compare the calibration of the QDR 4500A against criterion methods in a large heterogeneous population. DESIGN: DXA-derived body-composition data were obtained from 7 studies: 6 data sets were provided by the investigators, one of which was published. The data included fat mass and fat-free mass measured with a QDR 4500A and criteria measurements of body composition from total body water by dilution at 4 centers, densitometry from 1 center, and four-compartment analysis at 2 centers. RESULTS: In the cohort of 1195 subjects, 602 men and 593 women aged 19 82 y with a body mass index (in kg/m2) of 16-44, the fan-beam DXA overestimated fat-free mass (P < 0.05). A significant difference was observed in all 7 data sets, and the mean (+/-SE) was 5 +/- 1%. CONCLUSIONS: It is recommended that the lean soft tissue mass estimate with the fan-beam QDR 4500A be reduced by 5% and that for fat mass be increased by that same mass. This finding is particularly important because the National Health and Nutrition Examination Survey is using the QDR 4500A to assess body composition in a nationally representative sample of persons in the United States. PMID- 15883425 TI - Evening intake of alpha-lactalbumin increases plasma tryptophan availability and improves morning alertness and brain measures of attention. AB - BACKGROUND: Brain serotonin function is thought to promote sleep regulation and cognitive processes, whereas sleep abnormalities and subsequent behavioral decline are often attributed to deficient brain serotonin activity. Brain uptake of the serotonin precursor tryptophan is dependent on nutrients that influence the availability of tryptophan via a change in the ratio of plasma tryptophan to the sum of the other large neutral amino acids (Trp:LNAA). OBJECTIVE: We tested whether evening consumption of alpha-lactalbumin protein with an enriched tryptophan content of 4.8 g/100 g increases plasma Trp:LNAA and improves alertness and performance on the morning after sleep, particularly in subjects with sleep complaints. DESIGN: Healthy subjects with (n = 14) or without (n = 14) mild sleep complaints participated in a double-blind, placebo-controlled study. The subjects slept at the laboratory for 2 separate nights so that morning performance could be evaluated after an evening diet containing either tryptophan rich alpha-lactalbumin or tryptophan-low placebo protein. Evening dietary changes in plasma Trp:LNAA were measured. Behavioral (reaction time and errors) and brain measures of attention were recorded during a continuous performance task. RESULTS: Evening alpha-lactalbumin intake caused a 130% increase in Trp:LNAA before bedtime (P = 0.0001) and modestly but significantly reduced sleepiness (P = 0.013) and improved brain-sustained attention processes (P = 0.002) the following morning. Only in poor sleepers was this accompanied by improved behavioral performance (P = 0.05). CONCLUSION: Evening dietary increases in plasma tryptophan availability for uptake into the brain enhance sustained alertness early in the morning after an overnight sleep, most likely because of improved sleep. PMID- 15883426 TI - Genetic and environmental contributions to serum retinol and alpha-tocopherol concentrations: the Stanislas Family Study. AB - BACKGROUND: Although numerous environmental factors are documented to influence serum retinol and alpha-tocopherol concentrations, little is known about the genetic versus the environmental contributions to variations in these traits. OBJECTIVE: The aim of this study was to estimate additive genetic heritability and household effects for serum retinol and alpha-tocopherol concentrations in a variance component analysis. DESIGN: In a sample of 387 French families, information on serum retinol and alpha-tocopherol concentrations, usual dietary intake, lifestyle, and serum lipid profiles and related polymorphisms (apolipoprotein E, apolipoprotein C-III, apolipoprotein B, cholesteryl ester transfer protein, and lipoprotein lipase) was obtained. RESULTS: For serum retinol--after adjustment for sex, age, body mass index, alcohol consumption, oral contraceptive use, and serum albumin, triacylglycerol, and apolipoprotein A I concentrations--additive genetic effects and shared common environment contributed 30.5% and 14.2% of the total variance, respectively. For serum alpha tocopherol, approximately 22.1% of the total variance was due to the additive effects of genes and 18.7% to those of household environment, after adjustment for the covariates sex, age, vitamin E intake, oral contraceptive use, and cholesterol, triacylglycerol, and apolipoprotein A-I concentrations. For both vitamins, the influence of measured polymorphisms was not significant. Moreover, heritability and household effect estimates were not significantly different between the 4 classes of relatives and did not vary significantly when families shared more meals at home. CONCLUSIONS: The results show that serum retinol and alpha-tocopherol concentrations are under genetic control in healthy families. PMID- 15883427 TI - Low dietary zinc decreases erythrocyte carbonic anhydrase activities and impairs cardiorespiratory function in men during exercise. AB - BACKGROUND: The role of zinc in promoting physiologic function during exercise is not well understood. Although some zinc-containing enzymes are postulated to regulate energy expenditure, data are limited on the effect of restricted dietary zinc on metabolic responses during exercise. OBJECTIVE: This study determined the effects of low zinc intake on carbonic anhydrase activity in red blood cells (RBCs) and cardiorespiratory function during exercise. DESIGN: In this double blind, randomized crossover study, 14 men aged 20-31 y were fed low-zinc and supplemented (3.8 and 18.7 mg/d) diets made up of Western foods for 9-wk periods with a 6-wk washout. Peak work capacity, determined by using a cycle ergometer and a graded, progressive protocol, and a prolonged submaximal test (70% peak intensity for 45 min) were administered during the second and ninth weeks of each diet period. RESULTS: Dietary zinc did not affect hemoglobin or hematocrit. Low dietary zinc resulted in lower (P < 0.05) serum and erythrocyte zinc concentrations, zinc retention, and total carbonic anhydrase and isoform activities in RBCs. Peak oxygen uptake, carbon dioxide output, and respiratory exchange ratio were lower (P < 0.05), and ventilatory equivalents for metabolic responses during exercise were greater (P < 0.05), with low than with supplemental zinc intake. Similar functional responses were observed during prolonged, submaximal exercise. CONCLUSION: These findings indicate that low dietary zinc is associated with significant reductions in zinc status, including RBC carbonic anhydrase activities, and impaired metabolic responses during exercise. PMID- 15883428 TI - Lower plasma alpha-carboxyethyl-hydroxychroman after deuterium-labeled alpha tocopherol supplementation suggests decreased vitamin E metabolism in smokers. AB - BACKGROUND: Cigarette smoking increases the fractional disappearance rates of alpha-tocopherol and is associated with increased oxidative stress, but its effects on alpha-tocopherol metabolism are unknown. OBJECTIVE: We hypothesized that smokers would have less alpha-tocopherol available and consequently lower plasma alpha-carboxyethyl-hydroxychroman (alpha-CEHC), the alpha-tocopherol metabolite produced by a cytochrome P450-mediated process. DESIGN: Smokers and nonsmokers (n = 10 per group) were supplemented with deuterium-labeled alpha tocopheryl acetates (75 mg each d3-RRR-alpha-tocopheryl and d6-all-rac-alpha tocopheryl acetate) from day -6 to day -1, and plasma tocopherols and CEHCs were measured (day -6 through day 17). RESULTS: After 6 d of supplementation, plasma d3- and d6-alpha-tocopherol concentrations did not differ significantly between groups. Plasma d3- and d6-alpha-CEHCs were detectable only from day -5 to day 5. Before supplementation, unlabeled alpha- and gamma-CEHCs were approximately 60% and 40% lower, respectively, in smokers than in nonsmokers (P < or = 0.05). In addition, d0-, d3-, and d6-alpha-CEHC areas under the curves were approximately 50% lower in smokers (P < 0.05), and smokers had lower maximal d3-alpha-CEHC (P = 0.004) and d6-alpha-CEHC (P = 0.0006) concentrations. Notably, 2.9-4.7 times as much alpha-CEHC was produced from all-rac-alpha-tocopherol than from RRR-alpha tocopherol. During supplementation, smokers had about one-half (P < 0.05) the plasma total, d6-, or d3-alpha-CEHC concentrations that nonsmokers did given similar alpha-tocopherol concentrations. CONCLUSIONS: Smoking did not increase alpha-tocopherol disappearance through P450-mediated tocopherol metabolism. Therefore, the mechanism of increased alpha-tocopherol disappearance in smokers likely operates through oxidation pathways, which is consistent with alpha tocopherol's antioxidant function. Consequently, evaluating the molecular mechanism or mechanisms responsible for tocopherol metabolism under conditions of oxidative stress and the mechanisms that regulate alpha-tocopherol status is warranted. PMID- 15883429 TI - High prevalence of vitamin D deficiency among pregnant women and their newborns in northern India. AB - BACKGROUND: Vitamin D deficiency is prevalent in India, a finding that is unexpected in a tropical country with abundant sunshine. Vitamin D deficiency during pregnancy has important implications for the newborn and infant. There are few data from India about the prevalence of hypovitaminosis D in pregnancy and in the newborn. OBJECTIVE: Our aim was to determine the prevalence of osteomalacia and hypovitaminosis D in pregnancy and in cord blood and to correlate maternal 25 hydroxyvitamin D [25(OH)D] status with sun exposure, daily calcium intake (dietary plus supplemental), and intact parathyroid hormone (PTH) concentrations. DESIGN: Serum calcium, inorganic phosphorus, 25(OH)D, heat-labile alkaline phosphatase, and PTH were studied in 207 urban and rural pregnant subjects at term. Alkaline phosphatase and 25(OH)D were measured in the cord blood of 117 newborns. RESULTS: Mean maternal serum 25(OH)D was 14 +/- 9.3 ng/mL, and cord blood 25(OH)D was 8.4 +/- 5.7 ng/mL. PTH rose above the normal range when 25(OH)D was <22.5 ng/mL. Eighty-four percent of women (84.3% of urban and 83.6% of rural women) had 25(OH)D values below that cutoff. Fourteen percent of the subjects had elevated alkaline phosphatase (17% of urban and 7% of rural subjects). Calcium intake was uniformly low, although higher in urban (842 +/- 459 mg/d) than in rural (549 +/- 404 mg/d) subjects (P < 0.001). Maternal serum 25(OH)D correlated positively with cord blood 25(OH)D (r = 0.79, P < 0.001) and negatively with PTH (r = -0.35, P < 0.001). CONCLUSION: We observed a high prevalence of physiologically significant hypovitaminosis D among pregnant women and their newborns, the magnitude of which warrants public health intervention. PMID- 15883430 TI - Effects of chronic copper exposure during early life in rhesus monkeys. AB - BACKGROUND: Whether infants regulate copper absorption and the potential effects of excess copper in early life remain poorly defined. OBJECTIVE: The objective of the study was to assess copper retention, liver copper content, and liver function in infant rhesus monkeys fed infant formula containing 6.6 mg Cu/L. DESIGN: From birth to 5 mo of age, infant rhesus monkeys were fed formula that was supplemented with copper (0.6 mg Cu/L; n = 5) or not supplemented (n = 4). In all animals, weight and crown-rump length (by anthropometry), hemoglobin, hematocrit, plasma ceruloplasmin activity, and zinc and copper concentrations were measured monthly (birth to 6 mo) and at 8 and 12 mo. When the animals were 1, 5, and 8 mo old, liver copper and metallothionein concentrations, liver histology (by light and electron microscopy), and the number of Kupffer cells were assessed, and 67Cu retention was measured. Liver function was assessed by measurement of plasma alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, and alkaline phosphatase activities and protein, albumin, bilirubin, and blood urea nitrogen concentrations. RESULTS: 67Cu retention was 19.2% and 10.9% after 1 and 5 mo of copper treatment, respectively, compared with approximately 75% in controls at age 2 mo. At age 8 mo, 67Cu retention was 22.9% in copper-treated animals and 31.5% in controls. Liver histology remained normal by light microscopy, with mild ultrastructural signs of cell damage at 5 mo. Liver copper concentration was 4711, 1139, and 498 microg/g dry tissue at 1, 5, and 8 mo, respectively, in copper-treated animals and 250 microg/g at 2 mo in controls. Measurements could not be completed in all animals. CONCLUSIONS: No clinical evidence of copper toxicity was observed. Copper absorption was down regulated; increases in liver copper content at ages 1 and 5 mo did not result in histologic damage. Ultrastructural changes at age 5 mo could signal early cellular damage. PMID- 15883431 TI - S-equol, a potent ligand for estrogen receptor beta, is the exclusive enantiomeric form of the soy isoflavone metabolite produced by human intestinal bacterial flora. AB - BACKGROUND: The discovery of equol in human urine more than 2 decades ago and the finding that it is bacterially derived from daidzin, an isoflavone abundant in soy foods, led to the current nutritional interest in soy foods. Equol, unlike the soy isoflavones daidzein or genistein, has a chiral center and therefore can occur as 2 distinct diastereoisomers. OBJECTIVE: Because it was unclear which enantiomer was present in humans, our objectives were to characterize the exact structure of equol, to examine whether the S- and R-equol enantiomers are bioavailable, and to ascertain whether the differences in their conformational structure translate to significant differences in affinity for estrogen receptors. DESIGN: With the use of chiral-phase HPLC and mass spectrometry, equol was isolated from human urine and plasma, and its enantiomeric structure was defined. Human fecal flora were cultured in vitro and incubated with daidzein to ascertain the stereospecificity of the bacterial production of equol. The pharmacokinetics of S- and R- equol were determined in 3 healthy adults after single-bolus oral administration of both enantiomers, and the affinity of each equol enantiomer for estrogen receptors was measured. RESULTS: Our studies definitively establish S-equol as the exclusive product of human intestinal bacterial synthesis from soy isoflavones and also show that both enantiomers are bioavailable. S-equol has a high affinity for estrogen receptor beta (K(i) = 0.73 nmol/L), whereas R-equol is relatively inactive. CONCLUSIONS: Humans have acquired an ability to exclusively synthesize S-equol from the precursor soy isoflavone daidzein, and it is significant that, unlike R-equol, this enantiomer has a relatively high affinity for estrogen receptor beta. PMID- 15883432 TI - Beta-carotene-rich orange-fleshed sweet potato improves the vitamin A status of primary school children assessed with the modified-relative-dose-response test. AB - BACKGROUND: Beta-carotene-rich orange-fleshed sweet potato (OFSP) is an excellent source of provitamin A. In many developing countries, sweet potato is a secondary staple food and may play a role in controlling vitamin A deficiency. OBJECTIVE: The objective was to determine the efficacy of daily consumption of boiled and mashed OFSP in improving the vitamin A status of primary school children. DESIGN: Children aged 5-10 y were randomly assigned to 2 groups. The treatment group (n = 90) consumed 125 g boiled and mashed OFSP (1031 retinol activity equivalents/d as beta-carotene), and the control group (n = 90) consumed an equal amount of white fleshed sweet potato devoid of beta-carotene for 53 school days. All children were dewormed to exclude helminthic infection. The modified-relative-dose response test for vitamin A status was conducted before and after intervention. RESULTS: The estimated intervention effect for the ratio of 3,4-didehydroretinol to retinol (DR:R) was -0.008 (95% CI: -0.015, -0.001; P = 0.0203), which indicated a greater improvement in vitamin A liver stores in the treatment group than in the control group. The proportions of children with normal vitamin A status (DR:R < 0.060) in the treatment group tended to increase from 78% to 87% (P = 0.096) and did not change significantly (from 86% to 82%) in the control group (P = 0.267). These proportions were not used to test the intervention effect or within-group changes because the study was powered to test the intervention effect on DR:R. CONCLUSIONS: Consumption of OFSP improves vitamin A status and can play a significant role in developing countries as a viable long term food-based strategy for controlling vitamin A deficiency in children. PMID- 15883433 TI - Ascorbic acid supplementation and regular consumption of fresh orange juice increase the ascorbic acid content of human milk: studies in European and African lactating women. AB - BACKGROUND: Little is known about the influence of an increased intake of ascorbic acid (AA) on human milk AA output. OBJECTIVE: We aimed to compare human milk AA content in European and African women and to evaluate the influence of increased AA intake on human milk AA output. DESIGN: Apparently healthy lactating women were recruited. AA was analyzed by titration with 2,6-dichlorophenol indophenol. RESULTS: Mean human milk AA was approximately 50% lower (P < 0.001) in the African women (31 mg/kg; n = 171) than in the European women (63 mg/kg; n = 142). AA supplementation (1000 mg/d for 10 d) increased mean human milk AA from 19 to 60 mg/kg (P < 0.001) and from 60 to 70 mg/kg (P = 0.03) in 18 African and 10 European women, respectively. In 11 African women, mean human milk AA increased from 17 to 36 mg/kg (P < 0.001) after intake of 100 mg AA/d for 10 d. In African women, intake of 1 serving of orange juice per week had no significant effect, whereas 3 or 5 servings/wk ( approximately 100 mg AA/serving) for 6 wk increased mean human milk AA from 16 to 32 mg/kg (n = 13) and from 21 to 46 mg/kg (n = 13), respectively (P < 0.001). CONCLUSIONS: Human milk AA can be doubled or tripled by increased intake of AA in women with low human milk AA content at baseline. The response to a relatively high dose of AA was modest in European women in contrast with the 3-fold increase in mean human milk AA content in African women. These data indicate that human milk AA content is regulated. PMID- 15883434 TI - Supplementation of infant formula with long-chain polyunsaturated fatty acids does not influence the growth of term infants. AB - BACKGROUND: Adequate growth is an important indicator of health and well-being in infants. OBJECTIVE: Our objective was to determine the effect of supplementing infant formula with long-chain polyunsaturated fatty acids (LCPUFAs) on the growth of term infants. DESIGN: Using the methodology outlined by the Cochrane Collaboration, we reviewed all known randomized controlled trials that involved LCPUFA supplementation of infant formula fed to term infants. Outcome measures were weight, length, and head circumference. Original data obtained from the investigators of published trials were used. Outcomes were analyzed with fixed effects or random-effects model meta-analyses and were reported as weighted mean differences with 95% CIs. RESULTS: We identified 14 eligible trials that had data available for meta-analysis (1846 infants). Trial quality was generally high. Meta-analysis showed no significant effect of LCPUFA supplementation on infant weight, length, or head circumference at any assessment age. Similarly, subgroup analyses showed that supplementation with only n-3 LCPUFAs (no arachidonic acid) had no significant effect on infant weight, length, or head circumference. The source of LCPUFA supplementation (phospholipid or triacylglycerol) also did not significantly affect infant growth. CONCLUSION: We found no evidence that LCPUFA supplementation of infant formula influences the growth of term infants in either a positive or a negative way. PMID- 15883435 TI - Longitudinal changes in energy expenditure in girls from late childhood through midadolescence. AB - BACKGROUND: Longitudinal data on energy expenditure in children and adolescents are scarce. OBJECTIVE: The purpose of this study was to examine changes in energy expenditure and physical activity in girls from late childhood through midadolescence. DESIGN: We measured total energy expenditure (TEE) by doubly labeled water, resting metabolic rate (RMR) by indirect calorimetry, body composition by 18O dilution, and time spent in activity by an activity diary in 28 initially nonobese girls at approximately 10, approximately 12, and approximately 15 y of age. Changes with age in TEE, RMR, and activity energy expenditure (AEE), both in absolute terms and in adjusted analyses, and in physical activity level (PAL) and time spent sleeping, being sedentary, and in moderate and vigorous activity were evaluated by mixed-model repeated-measures analyses. RESULTS: Absolute TEE and AEE increased significantly from age 10 to age 15 y (P < 0.0001 for both). Absolute RMR at ages 12 and 15 y did not differ significantly, despite significant increases in fat-free mass and fat mass between the visits. PAL was significantly higher (P < 0.0001) at age 15 y than at age 10 or 12 y, whereas time spent being sedentary increased significantly from age 10 to age 15 y (P < 0.001), and AEE adjusted for fat-free mass appeared to decrease over the same interval. CONCLUSION: Conclusions drawn regarding changes with age in physical activity depend on the measure of physical activity assessed. PMID- 15883436 TI - Total homocysteine and its predictors in Dutch children. AB - BACKGROUND: Vitamin status, methylenentetrahydrofolate reductase (MTHFR) genotype, age, sex, and lifestyle factors are all predictors of total homocysteine (tHcy) concentrations in adults. Limited data are available about the influence of these factors on tHcy in children. OBJECTIVE: The objective was to describe tHcy and its predictors in Dutch children. DESIGN: A sample of 234 white children aged 0-19 y was analyzed cross-sectionally. RESULTS: The geometric mean tHcy concentrations were 5.1 (95% CI: 4.6, 5.6), 4.6 (4.2, 5.1), 6.2 (5.6, 6.9), 7.3 (6.7, 8.0), and 8.7 (7.9, 9.6) micromol/L in the 0-1, 2-5, 6-10, 11-14, and 15-19 y groups, respectively. Plasma folate and vitamin B-12 concentrations decreased markedly with age. The inverse association between tHcy and plasma folate seen at all ages was stronger than that between tHcy and plasma vitamin B 12. A negative association of plasma folate with tHcy was confined to folate concentrations <20 nmol/L. Homozygosity for the MTHFR 677C-->T polymorphism was identified in 8.2% of the children. The homocysteine concentration did not differ significantly between the MTHFR genotypes. CONCLUSIONS: This study provided age specific data regarding tHcy concentrations and their predictors in the whole range of childhood. The tHcy concentration increased as a function of age in both sexes. Plasma folate was a concentration-dependent predictor of tHcy. The MTHFR 677C-->T polymorphism played a minor role in determining tHcy concentrations in children. PMID- 15883437 TI - Diabetes mellitus worsens antioxidant status in patients with chronic pancreatitis. AB - BACKGROUND: Patients with chronic pancreatitis (CP) are at high risk of antioxidant deficiencies. Furthermore, this disease can lead to diabetes mellitus (DM) that could exacerbate the severity of oxidative stress. Oxidative stress and the resulting LDL oxidation are a major cause of atherosclerosis. OBJECTIVE: The objective of the study was to ascertain whether diabetes significantly modifies oxidative status in patients with CP. DESIGN: CP patients with or without DM were compared with type 1 DM patients and healthy control subjects. RESULTS: Two-way factorial analyses showed that a decrease in the plasma concentrations of vitamin A, vitamin E, and carotenoids accompanied both CP and DM, and CP was also associated with lower plasma concentrations of selenium and zinc, lower catalase activity, and higher plasma concentrations of copper. The lag phase of LDL oxidation was lower in CP patients with or without DM than in the control subjects, whereas there was no significant difference between type 1 DM patients and control subjects. Multivariate analysis showed that LDL vitamin E (R2 = 0.24, P < 0.0001) and fasting plasma glucose (R2 = 0.32, P < 0.0001) concentrations were the main determinants of the lag phase of LDL oxidation. CONCLUSIONS: Antioxidant status is altered in CP patients, particularly in those who also have DM. In these patients, a vitamin E deficiency and an elevated plasma glucose concentration were associated with significantly higher LDL oxidizability. PMID- 15883438 TI - Relation between liver fat content and the rate of VLDL apolipoprotein B-100 synthesis in children with protein-energy malnutrition. AB - BACKGROUND: Fatty infiltration of the liver is associated with an increased morbidity and mortality in children with severe protein-energy malnutrition (PEM), but its pathogenesis remains unclear. Although impaired synthesis of VLDL apolipoprotein B-100 (VLDL-apo B-100) is generally accepted as the pathogenetic mechanism, the rate of it synthesis has not been measured in children with PEM. OBJECTIVE: The objective of the study was to ascertain the relation between the degree of hepatic steatosis and the rate of VLDL-apo B-100 synthesis in children with PEM. DESIGN: The fractional and absolute rates of VLDL-apo B-100 synthesis were measured with a prime-constant intravenous infusion of [2H3]leucine in 13 severely malnourished children (8 boys and 5 girls) aged 7-18 mo. Hepatic fat content was estimated by computerized tomography scanning by using the ratio of liver to spleen (L:S) attenuation. The ratio is inversely related to hepatic fat content such that the lower the L:S, the greater the amount of fat in the liver. RESULTS: There were significant inverse relations between L:S attenuation and VLDL-apo B-100 concentration (P < 0.02), the absolute rate of VLDL-apo B-100 synthesis (P < 0.02), and plasma triacylglycerol (P < 0.02) and serum cholesterol (P < 0.05) concentrations. CONCLUSIONS: These results suggest that children with PEM synthesize VLDL-apo B-100 at a faster rate as the degree of hepatic fat infiltration increases. Thus, fatty infiltration of the liver in PEM is not due to a reduction in the synthesis of VLDL-apo B-100. PMID- 15883439 TI - A controlled 2-mo dietary fat reduction and soy food supplementation study in postmenopausal women. AB - BACKGROUND: Low intake of dietary fat and high intake of soy foods have been suggested to partly explain the lower breast cancer rates in Asia, perhaps because of lower endogenous estrogens. OBJECTIVE: The objective was to assess the hormonal and nonhormonal effects of diets resembling an Asian diet in terms of total fat and soy food contents. DESIGN: Fifty-seven postmenopausal women participated in a randomized, controlled, dietary intervention study. The subjects consumed a very-low-fat diet (VLFD; 11% of energy as fat), a Step I diet (25% of energy as fat) supplemented with soy food (SFD; 50 mg isoflavones/d), or a control Step I diet (CD; 27% of energy as fat) with no soy food. All diets were prepared at the General Clinical Research Center of the University of Southern California. Serum hormones and other markers were measured at baseline and every 2 wk during the 8 wk of intervention. RESULTS: There were no significant differences in total estradiol and sex hormone binding globulin at the completion of the intervention between women in the SFD and VLFD groups and those in the CD group. Serum insulin decreased significantly in the SFD group, and leptin decreased significantly in the SFD and VLFD groups; however, these changes did not differ significantly from the changes in the CD group. CONCLUSIONS: This study does not provide evidence that ingestion of soy food or a VLFD significantly reduces estrogen concentrations in postmenopausal women. However, short-term changes in diet may have significant and beneficial effects on blood insulin and leptin concentrations. PMID- 15883440 TI - Plasma arginine concentrations are reduced in cancer patients: evidence for arginine deficiency? AB - BACKGROUND: The disturbances leading to cancer cachexia remain to be unraveled. Preliminary evidence suggests that arginine availability in cancer is reduced. However, no valid data are available on plasma arginine concentrations in cancer patients. OBJECTIVE: We aimed to determine whether there is evidence for disturbed arginine metabolism in cancer. DESIGN: We measured plasma arginine concentrations postabsorptively in patients with various types of tumors, hypothesizing that arginine concentrations would be lower than those in age- and sex-matched control subjects. Patients with localized tumors with a range of metabolic implications were studied: breast cancer (no weight loss), colonic cancer (sometimes weight loss), and pancreatic cancer (usually weight loss). Plasma amino acid concentrations were measured by HPLC. RESULTS: Plasma arginine concentrations were lower in patients with cancer (breast cancer: 80 +/- 3 compared with 103 +/- 9 micromol/L; colonic cancer: 80 +/- 3 compared with 96 +/- 7 micromol/L; pancreatic cancer: 76 +/- 5 compared with 99 +/- 7 micromol/L; P < 0.05 versus respective age- and sex-matched control subjects), irrespective of tumor type, weight loss, tumor stage, or body mass index (correlations with P > 0.05). CONCLUSIONS: Malignant tumors associated with various degrees of metabolic derangements are all associated with decreased plasma arginine concentrations, even without weight loss. This suggests that decreased arginine availability is a specific feature of the presence of cancer. These disturbances in arginine metabolism could contribute to the cascade of metabolic events leading to cancer cachexia. PMID- 15883441 TI - Dairy, calcium, and vitamin D intakes and prostate cancer risk in the National Health and Nutrition Examination Epidemiologic Follow-up Study cohort. AB - BACKGROUND: Dairy intake may increase prostate cancer risk, but whether this is due to calcium's suppression of circulating vitamin D remains unclear. Findings on calcium and vitamin D intake and prostate cancer are inconsistent. OBJECTIVE: We examined the association of dairy, calcium, and vitamin D intake with prostate cancer. DESIGN: In a prospective study of 3612 men followed from 1982-1984 to 1992 for the first National Health and Nutrition Examination Epidemiologic Follow up Study, 131 prostate cancer cases were identified. Dietary intake was estimated from questionnaires completed in 1982-1984. Relative risk (RR) and 95% CIs were estimated by using Cox proportional hazards models adjusted for age, race, and other covariates. RESULTS: Compared with men in the lowest tertile for dairy food intake, men in the highest tertile had a relative risk (RR) of 2.2 (95% CI: 1.2, 3.9; trend P = 0.05). Low-fat milk was associated with increased risk (RR = 1.5; 95% CI: 1.1, 2.2; third compared with first tertile; trend P = 0.02), but whole milk was not (RR = 0.8; 95% CI: 0.5, 1.3; third compared with first tertile; trend P = 0.35). Dietary calcium was also strongly associated with increased risk (RR = 2.2; 95% CI: 1.4, 3.5; third compared with first tertile; trend P = 0.001). After adjustment for calcium intake, neither vitamin D nor phosphorus was clearly associated with risk. CONCLUSIONS: Dairy consumption may increase prostate cancer risk through a calcium-related pathway. Calcium and low-fat milk have been promoted to reduce risk of osteoporosis and colon cancer. Therefore, the mechanisms by which dairy and calcium might increase prostate cancer risk should be clarified and confirmed. PMID- 15883442 TI - Significant correlations of plasma homocysteine and serum methylmalonic acid with movement and cognitive performance in elderly subjects but no improvement from short-term vitamin therapy: a placebo-controlled randomized study. AB - BACKGROUND: Deficiencies of vitamin B-12, folic acid, and vitamin B-6-as defined by laboratory measures-occur in 10-20% of elderly subjects. The clinical significance remains unresolved. OBJECTIVE: The objective was to explore any association between vitamin status and vitamin treatment and movement and cognitive performance in elderly subjects. DESIGN: Community-dwelling subjects (n = 209) with a median age of 76 y were randomly assigned to daily oral treatment with 0.5 mg cyanocobalamin, 0.8 mg folic acid, and 3 mg vitamin B-6 or placebo (double blind) for 4 mo. Movement and cognitive performance tests were performed before and after treatment. RESULTS: A high plasma total homocysteine (tHcy) concentration (> or =16 micromol/L) was found in 64% of men and in 45% of women, and a high serum methylmalonic acid (MMA) concentration (> or =0.34 micromol/L) was found in 11% of both sexes. Movement time, digit symbol, and block design (adjusted for age, sex, smoking, and creatinine) correlated independently with plasma tHcy (P < 0.01, < 0.05, and < 0.01, respectively); the simultaneity index and block design correlated with serum MMA (P < 0.05 for both). Vitamin therapy significantly decreased plasma tHcy (32%) and serum MMA (14%). No improvements were found in the movement or cognitive tests compared with placebo. Neither vitamin therapy nor changes in plasma tHcy, serum MMA, serum vitamin B-12, plasma folate, or whole-blood folate correlated with changes in movement or cognitive performance. CONCLUSIONS: High plasma tHcy and serum MMA were prevalent and correlated inversely with movement and cognitive performance. Oral B vitamin treatment normalized plasma tHcy and serum MMA concentrations but did not affect movement or cognitive performance. This might have been due to irreversible or vitamin-independent neurocognitive decline or to an insufficient dose or duration of vitamins. PMID- 15883443 TI - Association of diet with serum insulin-like growth factor I in middle-aged and elderly men. AB - BACKGROUND: Insulin-like growth factor I (IGF-I) has been implicated in several chronic diseases, including cancer, heart disease, and osteoporosis. OBJECTIVE: Our aim was to assess whether intakes of total energy, alcohol, vitamins, minerals, and foods rich in protein and minerals (including red meat, fish and seafood, poultry, and milk) are associated with serum IGF-I concentrations in middle-aged and elderly men. DESIGN: We measured serum IGF-I concentrations in 226 free-living healthy men aged 42-76 y. The average of fourteen 24-h dietary telephone interviews performed over 1 y was used to estimate long-term dietary intake. RESULTS: We observed statistically significant positive associations between intakes of protein (P for trend = 0.001) and zinc (P for trend = 0.002) and serum IGF-I concentrations after adjusting for age. The difference in mean IGF-I concentrations for the highest compared with the lowest quintile of intake was approximately 17% (162 microg/L compared with 139 microg/L) for protein and approximately 16% (166 microg/L compared with 143 microg/L) for zinc. Consumption of red meat (P for trend = 0.05) and fish and seafood (P for trend = 0.07) was modestly positively associated with IGF-I concentrations. Other dietary factors were not associated with IGF-I concentrations. CONCLUSION: In this population of healthy well-nourished men, greater dietary intakes of protein, zinc, red meat, and fish and seafood were associated with higher IGF-I concentrations. PMID- 15883444 TI - Effects of school milk intervention on cortical bone accretion and indicators relevant to bone metabolism in Chinese girls aged 10-12 y in Beijing. AB - BACKGROUND: We previously reported that increased milk consumption enhances growth and bone mineral accretion in Chinese girls aged 10-12 y. OBJECTIVE: Our objective was to evaluate the effects of milk supplementation on cortical bone accretion and to study the physiologic mechanisms underlying the observed changes in bone. DESIGN: Chinese girls aged 10 y were randomly assigned into calcium fortified milk (Ca milk), calcium and vitamin D-fortified milk (CaD milk), and control groups according to their schools in a 24-mo school milk intervention trial. Periosteal and medullary diameters of metacarpal bone were measured at baseline and 24 mo in the Ca milk (n = 177), CaD milk (n = 210), and control (n = 219) groups. Insulin-like growth factor I (IGF-I), parathyroid hormone (PTH), bone alkaline phosphatase (BAP), osteocalcin, and deoxypyridinoline concentrations were measured at baseline and at 12 and 24 mo in the Ca milk (n = 43), CaD milk (n = 44), and control (n = 41) groups. RESULTS: After adjustment for pubertal status and clustering by school, 24-mo supplementation led to greater increases in periosteal diameter (1.2%) and cortical thickness (5.7%) and to smaller gains in medullary diameter (6.7%) than did the control (P < 0.05). The CaD milk group had lower serum BAP at 12 mo (19.9%) and lower serum PTH at 12 (46.2%) and 24 (16.4%) mo than did the control group (P < 0.05). The effect of milk supplementation on increasing IGF-I concentrations at 24 mo (16.7-23.3%) was significant in individual analyses but not after adjustment for clustering by school. CONCLUSIONS: Milk supplementation showed positive effects on periosteal and endosteal apposition of cortical bone. PMID- 15883445 TI - Quercetin, fruit consumption, and bone mineral density. PMID- 15883446 TI - Vitamin D deficiency during pregnancy: a risk factor not only for fetal growth and bone metabolism but also for correct development of the fetal immune system? PMID- 15883447 TI - Absorption of iron from ferritin. PMID- 15883448 TI - Hormonal and lifestyle determinants of appendicular skeletal muscle mass in men: the MINOS Study. PMID- 15883450 TI - A life cycle micronutrient perspective for women's health. AB - Micronutrients not only benefit women's health during childbearing years and during pregnancy and lactation, but they also have substantial impact on women's health during adolescence and the aging years. Thus, for women, diet quality is important for health today and in the future. Realizing that there are many ways to improve the quality of a diet and to obtain adequate amounts of vitamins and minerals from foods, food-based approaches are still not attaining adequate intakes in most women, both in the United States and worldwide. Efforts are needed to improve diet quality, focusing on the diet as a whole, and not on single vitamins or minerals. However, consideration must be given to fortified foods and/or supplements to insure micronutrient adequacy. PMID- 15883451 TI - Dietary Reference Intakes: development and uses for assessment of micronutrient status of women--a global perspective. AB - This paper reviews the process of developing the Dietary Reference Intakes (DRIs) and provides a synopsis of the micronutrient status of women worldwide. At a 1993 symposium held by the Food and Nutrition Board (FNB) of the Institute of Medicine (IOM), it was decided that the Recommended Dietary Allowances (RDAs) would be replaced by the DRIs, which would address several issues that the RDAs did not, including chronic disease risk reduction, upper levels for nutrients where toxicity data existed, and the possible health benefits of some food components that did not meet the traditional definition of a nutrient. Another important distinction is that because the DRIs are comprised of 4 reference values -the Estimated Average Requirement (EAR), RDA, Adequate Intake (AI), and a tolerable Upper Level (UL) -and not a single reference value like the previous RDAs, they could be used to differentiate planning from diagnosis or assessment. The latest DRIs and nutrient intakes are shown for iron, zinc, calcium, Vitamin A and folate status in women in the United States. Data on the micronutrient status of women globally are much more limited. Summary statistics on iron deficiency anemia, night blindness, and risk of zinc deficiency are summarized. PMID- 15883452 TI - Micronutrient deficiencies and gender: social and economic costs. AB - Vitamin and mineral deficiencies adversely affect a third of the world's people. Consequently, a series of global goals and a serious amount of donor and national resources have been directed at such micronutrient deficiencies. Drawing on the extensive experience of the authors in a variety of institutional settings, the article used a computer search of the published scientific literature of the topic, supplemented by reports and published and unpublished work from the various agencies. In examining the effect of sex on the economic and social costs of micronutrient deficiencies, the paper found that: (1) micronutrient deficiencies affect global health outcomes; (2) micronutrient deficiencies incur substantial economic costs; (3) health and nutrition outcomes are affected by sex; (4) micronutrient deficiencies are affected by sex, but this is often culturally specific; and finally, (5) the social and economic costs of micronutrient deficiencies, with particular reference to women and female adolescents and children, are likely to be considerable but are not well quantified. Given the potential impact on reducing infant and child mortality, reducing maternal mortality, and enhancing neuro-intellectual development and growth, the right of women and children to adequate food and nutrition should more explicitly reflect their special requirements in terms of micronutrients. The positive impact of alleviating micronutrient malnutrition on physical activity, education and productivity, and hence on national economies suggests that there is also an urgent need for increased effort to demonstrate the cost of these deficiencies, as well as the benefits of addressing them, especially compared with other health and nutrition interventions. PMID- 15883453 TI - Multiple micronutrients in pregnancy and lactation: an overview. AB - This overview of multiple micronutrients during pregnancy and lactation emphasizes 2 relatively neglected issues. The first is that maternal micronutrient status in the periconceptional period, and throughout pregnancy and lactation, should be viewed as a continuum; too often these 3 stages are treated and discussed separately from both a scientific and a public health perspective. Iron and vitamin B-12 are included as examples to stress how status at conception affects maternal, fetal, and infant status and health until the child is weaned. The second issue is that while most attention has been focused on a few micronutrients, for example iron and folate as discussed elsewhere in this Supplement, multiple micronutrient deficiencies occur simultaneously when diets are poor. Some of these deserve more attention as causes of poor pregnancy outcome, including other B vitamin deficiencies that result in homocysteinemia, antioxidants, vitamin D, and iodine. In lactation, maternal status or intake of the B vitamins (except folate), vitamin A, selenium and iodine strongly affect the amount of these nutrients secreted in breast milk. This can result in the infant consuming substantially less than the recommended amounts and further depleting stores that were low at birth. While the optimal mode of meeting recommended micronutrient intakes is an adequate diet, in some situations supplementation is also important. Unfortunately, information is lacking on the optimal formulation of micronutrient supplements for pregnant women, and the need to continue these supplements during lactation is not recognized in many situations where maternal and infant health could benefit. PMID- 15883454 TI - Folic acid supplementation and the occurrence of congenital heart defects, orofacial clefts, multiple births, and miscarriage. AB - Key research findings relative to the question of whether maternal use of folic acid before and during pregnancy reduces the chance that offspring will be born with a congenital heart defect or an orofacial cleft are reviewed in this paper. Observational studies in general support an association between maternal use of multivitamins containing folic acid and a reduction in the occurrence of congenital heart defects and orofacial clefts. Results from one randomized controlled trial (RCT) provide the strongest evidence that multivitamins prevent congenital heart defects, but this RCT did not provide evidence that multivitamins prevent orofacial clefts. In addition, most observational and interventional studies are not designed to detect an independent effect from folic acid. Early studies suggested that periconceptional multivitamin use was associated with an increased occurrence of both miscarriages and multiple births, which has resulted in a great deal of controversy about the safety of folic acid use during pregnancy. We also review reports that were designed to answer these questions with more definitive data. When more substantial evidence about the effect of periconceptional folic acid on the occurrence of congenital heart defects and orofacial clefts is reported, we will have additional support for promoting folic acid intervention programs. All women capable of becoming pregnant should continue to consume 400 mug/d of folic acid in addition to a healthy diet as advised. PMID- 15883455 TI - Iron status during pregnancy: setting the stage for mother and infant. AB - Supplementation with iron is generally recommended during pregnancy to meet the iron needs of both mother and fetus. When detected early in pregnancy, iron deficiency anemia (IDA) is associated with a > 2-fold increase in the risk of preterm delivery. Maternal anemia when diagnosed before midpregnancy is also associated with an increased risk of preterm birth. Results of recent randomized clinical trials in the United States and in Nepal that involved early supplementation with iron showed some reduction in risk of low birth weight or preterm low birth weight, but not preterm delivery. During the 3rd trimester, maternal anemia usually is not associated with increased risk of adverse pregnancy outcomes and may be an indicator of an expanded maternal plasma volume. High levels of hemoglobin, hematocrit, and ferritin are associated with an increased risk of fetal growth restriction, preterm delivery, and preeclampsia. While iron supplementation increases maternal iron status and stores, factors that underlie adverse pregnancy outcome are considered to result in this association, not iron supplements. On the other hand, iron supplements and increased iron stores have recently been linked to maternal complications (eg, gestational diabetes) and increased oxidative stress during pregnancy. Consequently, while iron supplementation may improve pregnancy outcome when the mother is iron deficient it is also possible that prophylactic supplementation may increase risk when the mother does not have iron deficiency or IDA. Anemia and IDA are not synonymous, even among low-income minority women in their reproductive years. PMID- 15883456 TI - Menopause, micronutrients, and hormone therapy. AB - Micronutrient and herbal/phytochemical supplements are of increasing interest as potential alternatives to using estrogen therapy in treating menopausal symptoms. This article provides an overview of the questionnaires that assess menopausal symptoms and research efforts to better standardize symptom assessment. The reported rate of symptoms varies by ethnicity, stage of menopause, hormonal therapy and the measurement method. The use of estrogen therapy has declined sharply after the Women's Health Initiative (WHI) Hormone Trial was stopped early because the potential risks outweighed potential benefits. There is a limited research base that addresses the efficacy of supplements in controlling menopausal symptoms. The generalizability of several studies is limited because the study participants experiences menopause as the results of treatment for breast cancer. The article concludes with a review of guidelines and of issues that need to be addressed in future research studies with emphasis on questions related to clinical practice. PMID- 15883457 TI - Osteoporosis: the role of micronutrients. AB - Osteoporosis and low bone mass are currently estimated to be a major public health threat. Adequate nutrition plays a major role in the prevention and treatment of osteoporosis; the micronutrients of greatest importance are calcium and vitamin D. Calcium has been shown to have beneficial effects on bone mass at all ages, although the results are not always consistent. Higher doses than the current US recommendation (600 IU) of vitamin D in the elderly (age > or = 65 y) may actually be required for optimal bone health (800-1000 IU/d). The elderly can clearly benefit from increased vitamin D intakes; however, the potential importance of vitamin D in peak bone mass is just being investigated. Vitamin D has been related to falls, with supplementation reducing the number of falls. There are clear fracture benefits demonstrated in randomized clinical trials of calcium and vitamin D supplementation. The other micronutrient needs for optimizing bone health can be easily met by a healthy diet that is high in fruits and vegetables to ensure adequate intakes for magnesium, potassium, vitamin C, vitamin K, and other potentially important nutrients. Healthcare professionals need to be aware of the importance of adequate calcium and vitamin D intakes (easily monitored by serum 25(OH)D) for optimal bone health, as well as the prevention of falls and fractures. In addition, a healthy diet that includes 5 servings a day of fruits and vegetables should optimize the intake of micronutrients required for bone health. PMID- 15883458 TI - Micronutrient requirements in older women. AB - The nutritional requirements of older women is an area of great interest because the extended life expectancy leads to an increase in women living into their 80s, 90s, and longer. The recommended dietary allowances (RDAs) and dietary reference intakes (DRIs) are not specific for women living to advanced ages, and little research has been conducted specifically on the micronutrient needs of elderly women. Older adults are at greater risk for nutritional deficiencies than are younger adults due to physiologic changes associated with aging, acute and chronic illnesses, prescription and over-the-counter medications, financial and social status, and functional decline. Among the significant age-associated changes in nutrient requirements, the need for energy decreases and the requirements for protein increase with age. Among the micronutrients, the significant ones that may be associated with deficiencies in elderly women include vitamin B-12, vitamin A, vitamin C, vitamin D, calcium, iron, zinc, and other trace minerals. In old and very old women, these are micronutrients of interest but there is a great need for research to determine appropriate recommendations. The importance of these selected nutrients and the reasons for the likelihood of deficiency are discussed briefly. However, there is little specific information regarding micronutrient requirements for elderly women. One reason for this is the difficulty in conducting reliable and valid studies due to the heterogeneity of older adults and their unique rate of aging associated with their health status, limited income, disability, and living situation. PMID- 15883459 TI - Micronutrient requirements of physically active women: what can we learn from iron? AB - The health benefits of physical activity are well established and there is increasing recognition of the importance of fitness as a key modulator of chronic disease. The impact of physical activity on micronutrient requirements is a topic of tremendous interest to the lay public, but the interest is in sharp contrast to data from well-designed studies. Research in this area is poorly controlled for nutritional status of the participants, standardized exercise protocols, markers and cutoff points for measurement of micronutrient status, and variability in subject characteristics. The micronutrient status of women in the general population is of concern, but it is not clear that physical activity increases the requirement of most micronutrients. When dietary intake is adequate, the results of most studies are either equivocal or show no benefit to performance of supplementation. In the few instances where exercise does appear to increase an individual's requirement, the increase can be obtained within the additional calories required for energy balance. In the absence of consistent data, micronutrient supplementation is often indiscriminate without regard to nutrient status. Because iron is such a key nutrient for physical activity, and the status in women is often compromised, it serves as a useful example of how current research limits the ability to make recommendations regarding the impact of exercise on micronutrients requirements in women. With the recent recognition of the importance of physical activity to the prevention and treatment of chronic diseases through the life span, more attention should be focused on the impact of exercise on micronutrient requirements, especially in the context of weight loss regimens. PMID- 15883460 TI - Educating the next generation of healers. PMID- 15883461 TI - Spiritual struggle: effect on quality of life and life satisfaction in women with breast cancer. AB - BACKGROUND: Women with breast cancer experience stressors affecting quality of life (QOL) and life satisfaction. Little is known about effects of spiritual struggle as a coping strategy on QOL and life satisfaction. PURPOSE: Examine relationships between spiritual struggle, QOL, and life satisfaction. METHOD: Nonprobablility sample of 100 participants recruited from an Internet Web site with mailed questionnaires. Three instruments were used: breast cancer-specific version of Functional Assessment of Cancer Therapy Scale (FACT-B), Functional Assessment of Chronic Illness Therapy-Spiritual (FACIT-Sp-12) combined for QOL, Negative Coping subscale of Religious Coping (RCOPE) for spiritual struggle, and a single-item measuring life satisfaction. FINDINGS: Small inverse relationships between spiritual struggle, QOL (r = -.36, p < .001), and life satisfaction (r = .31, p < .001) existed. CONCLUSIONS: Spiritual struggle gives voice to women's questionings implying lower QOL and life satisfaction. IMPLICATIONS: Assessment of and assistance with managing spiritual struggle are necessary to promote QOL and life satisfaction among those facing difficult health problems. PMID- 15883463 TI - The Spirituality Scale: development and psychometric testing of a holistic instrument to assess the human spiritual dimension. AB - PURPOSE: The purpose of this study was to develop, refine, and evaluate the psychometric characteristics of the Spirituality Scale (SS). The SS is a holistic instrument that attempts to measure the beliefs, intuitions, lifestyle choices, practices, and rituals representative of the human spiritual dimension and is designed to guide spiritual interventions. METHOD: A researcher-developed instrument was designed to assess spirituality from a holistic perspective. Items were generated to measure four conceptualized domains of spirituality. The SS was completed by 240 adults with chronic illness. FINDINGS: Psychometric analysis of the SS provided strong evidence of the reliability and validity of the instrument. Three factors of spirituality that supported the theoretical framework were identified: Self-Discovery, Relationships, and Eco-Awareness. IMPLICATIONS: These findings can assist in facilitating the inclusion of spirituality in health care and have the potential to provide a transforming vision for nursing care and a vehicle to evoking optimal patient outcomes. PMID- 15883465 TI - Meaning in life and psycho-spiritual functioning: a comparison of breast cancer survivors and healthy women. AB - PURPOSE: Incorporating holistic health perspectives, this study compared and examined relationships among meaning in life, spirituality, perceived stress, and psychological distress in breast cancer survivors (BCS) and healthy women. METHODS: Standardized self-report measures were completed once by all participants (N = 78). FINDINGS: Group comparison revealed statistically significant variances across the measures. Covariate analysis identified BCS without children had less meaningful lives and greater stress and distress than BCS with children and participants without cancer. Significant correlations (p > .001) between meaning in life and spirituality (r = .43), stress (r = -.39), and distress (r = -.41) were also identified. CONCLUSION: Personal factors (i.e., being a parent) may be especially important in BCS. Also, psychological and spiritual variables are highly correlated, suggesting the use of an integrated term psycho-spiritual functioning. IMPLICATIONS: Holistic nursing interventions can facilitate self-awareness, interpersonal connection, and living a meaningful life, particularly in vulnerable patients such as BCS without children. PMID- 15883466 TI - Continued encounters: the experience of after-death communication. AB - PURPOSE: To analyze and categorize the various forms of after-death communication (ADC) and describe the effects on the bereaved. METHOD: In this qualitative descriptive study of 9 men and 9 women, data were collected during in-depth interviews using the Grief and Mourning Status Interview and Inventory and semistructured interview questions. Transcripts of the interviews were analyzed and coded for content. FINDINGS: Four categories of ADCs were identified: (a) visions and dreams, (b) lost-things-found, (c) symbolic messages, and (d) sightings. Both men and women experience ADCs; however, women are more likely to discuss the events with others. CONCLUSION: Although not everyone encounters or recognizes the phenomenon of ADC, it is a common aspect of the bereavement experience. IMPLICATIONS: Nurses and other health care providers have an important role in supporting and educating the public, especially the bereaved, regarding the phenomenon of ADC. PMID- 15883467 TI - Assessing clients' sexual health as a component of holistic nursing practice: senior nursing students share their experiences. AB - A phenomenological study was conducted to investigate baccalaureate nursing students' experiences with assessing the sexual health of their clients. The participants were 10 female nursing students. The participants were asked to respond to the statement: Tell me about your experiences with assessing your clients' sexual health and concerns. Data were analyzed using Van Manen's criteria for analysis. The common themes were (a) recognized sexual assessment as a component of holistic nursing, (b) experienced discomfort exploring clients' sexual health, (c) believed that sexual assessment was warranted only if related to the diagnosis, and (d) held that the assessment was less important than other assessments. Information from this study has potential to inform nurse educators engaged in clinical teaching and curriculum development that sexual health assessment must be valued as a study area if holistic nursing care is to be a part of professional nursing practice. PMID- 15883469 TI - Spirituality and self-management of diabetes in African Americans. AB - Attention to spirituality is especially important for nurses when providing care to African Americans. Spirituality is deeply embedded in their rich cultural heritage. For many African Americans, spirituality is intertwined into all aspects of life, including beliefs about health and illness. Therefore, it is imperative that nurses understand the relationship between African American spirituality, health, and self-management of illness to provide culturally competent care to African Americans. The purpose of this article is to summarize the research literature on African American spirituality, health, and self management as it relates to Type 2 diabetes, an illness that involves complex self-care management. Recommendations for holistic nursing practice and research related to this literature are also identified. PMID- 15883471 TI - Does the pancreas really produce much more lipase than required for fat digestion? AB - Thirty years ago, it was reported that a linear relationship does not exist between the amounts of human pancreatic lipase secreted in chronic pancreatitis and the degree of steatorrhea, which was considered to appear only after more than 90% of the pancreatic secretory capacity had been lost. From these observations, it was generally thought that the lipolytic potential of the pancreas is much higher than required. In recent years, however, it has been noted that: 1) the level of inhibition of digestive lipases and gastrointestinal lipolysis by the lipase inhibitor orlistat were almost linearly correlated with the amount of excreted fat; 2) in minipigs with experimentally-induced pancreatic exocrine insufficiency, the amounts of enteric-coated pancreatic extracts needed for restoring fat digestion to normal levels were estimated to be much higher than those usually administered; 3) human pancreatic lipase specific activity on meal triglycerides is 3 orders of magnitude lower than the very high specific activity usually measured under experimental in vitro conditions which are far from physiological conditions; 4) in patients with reduced human pancreatic lipase secretion, gastric lipase plays a significant role in fat digestion. This last observation might explain the absence of a linear relationship between human pancreatic lipase secretion in chronic pancreatitis and steatorrhea. From the low specific activity displayed by human pancreatic lipase on meal triglycerides, one can better understand why more lipase than expected is needed, why fat digestion lasts for more than a few minutes and, finally, why there is not such an excess secretory capacity for lipase as had been previously thought. PMID- 15883472 TI - Chemoradiation for ductal pancreatic carcinoma: principles of combining chemotherapy with radiation, definition of target volume and radiation dose. AB - Review of the role of chemoradiotherapy in the treatment of locally advanced pancreatic cancer with a specific focus on the technical feasibility and the integration of chemoradiotherapy into multimodal treatment concepts. Combined chemoradiotherapy of pancreatic cancer is a safe treatment with an acceptable profile of side effects when applied with modern planning and radiation techniques as well as considering tissue tolerance. Conventionally fractionated radiation regimens with total doses of 45-50 Gy and small-volume boost radiation with 5.4 Gy have found the greatest acceptance. Locoregional lymphatic drainage should be included in the planning of target volumes because the risk of tumor involvement and local or loco-regional recurrence is high. Up to now, 5 fluorouracil has been considered the "standard" agent for concurrent chemoradiotherapy. The role of gemcitabine given concurrently with radiation has not yet been defined, since high local efficacy may also be accompanied by enhanced toxicities. In addition, no dose or administration form has been determined to be "standard" up to now. The focus of presently ongoing research is to define an effective and feasible regimen of concurrent chemoradiotherapy. While preliminary results indicate promising results using gemcitabine-based chemoradiotherapy, reliable data derived from mature phase III trials are greatly needed. Intensity-modulated radiotherapy has been developed to improve target specific radiation and to reduce organ toxicity. Its clinical relevance still needs to be defined. PMID- 15883473 TI - Estimating the diagnostic accuracy of procalcitonin as a marker of the severity of acute pancreatitis: a meta-analytic approach. AB - CONTEXT: Approximately 15-20% of cases of acute pancreatitis are categorized as severe. There is a lack of accurate predictors of disease severity. Several studies have evaluated the usefulness of procalcitonin as a marker of severe disease. Reports regarding the diagnostic accuracy of procalcitonin are conflicting. OBJECTIVE: The present meta-analysis was carried out to evaluate the relevance of procalcitonin as a predictor of disease severity. METHODS: Two investigators working independently attempted to locate eligible studies by electronic and manual means. Studies in which at least one of the markers of disease severity was procalcitonin were included for analysis. For all the studies included, the following parameters were calculated: true positive, false negative, false positive and true negative. A summary receiver operating characteristic (SROC) curve was generated from these parameters. RESULTS: Four studies were finally included in the analysis. The unweighted regression line parameters b and i were 3.633 and 1.399, respectively. The values for b and i for weighted regression line were 3.637 and 1.428. The SROC curve generated demonstrated that procalcitonin is not a good predictor of the severity of acute pancreatitis. CONCLUSION: The available data indicates that procalcitonin cannot be considered a good marker for assessing the severity of pancreatitis. PMID- 15883474 TI - Mutations in MODY genes are not common cause of early-onset type 2 diabetes in Mexican families. AB - CONTEXT: Maturity-onset diabetes of the young (MODY) is a monogenic form of diabetes mellitus characterized by autosomal dominant inheritance, early age of onset and a primary insulin secretion defect. Certain MODY gene sequence variants may be involved in polygenic forms of type 2 diabetes. OBJECTIVE: We assessed the contribution of MODY genes to the etiology of type 2 early-onset diabetes in 23 Mexican families, including five with apparently autosomal dominant inheritance. PATIENTS: Twenty-three unrelated Mexican families with early-onset type 2 diabetes previously screened for the presence of glucokinase mutations, were studied. DESIGN: We screened MODY genes for sequence variants by PCR-SSCP analysis and automated sequencing. We performed a functional analysis of the HNF 1alpha P379H recombinant protein in vitro in both HeLa and RINm5f beta-cell lines. MAIN OUTCOME MEASURES: MODY gene mutation screening and P379H mutant protein transactivation assay. RESULTS: No mutations were detected in the HNF 4alpha, IPF-1, NEUROD1 or HNF-1beta genes in any of the families studied. A new mutation (P379H) of the HNF-1alpha gene was identified in one MODY family. RINm5f and HeLa cell transfection assays revealed decreased transactivation activity of the mutant protein on the human insulin promoter. CONCLUSIONS: All known MODY genes were screened for abnormalities in this cohort of early-onset diabetes families which included 5 MODY pedigrees. We identified a new HNF-1alpha MODY mutation (P379H) and demonstrated that it reduces the transactivation potential of the mutant protein on the human insulin promoter. No other mutation was identified in this cohort indicating that abnormalities in MODY genes are generally not a common cause of early-onset diabetes and this includes MODY families in Mexico. PMID- 15883475 TI - Impact of ethnicity on outcome in pancreatic carcinoma. AB - CONTEXT: Ethnicity is an important factor known to impact the biology and outcome of many cancers. In pancreatic cancer, African Americans are thought to have a higher incidence and poorer prognosis than Whites. PATIENTS: A total 645 pancreatic cancer patients were identified in the database, of which, 530 patients were eligible for this study and retained for the statistical analysis. Of the 530 patients, 137 (25.8%) were Blacks, 393 (74.2%) were Whites, 275 (51.9%) were males and 255 (48.1) were females. Ages ranged from 21 to 93 years with values of 62.3+/-13.3 years, 64.2+/-11.6 years, 63.1+/-10.9 and 64.3+/-13.3 (mean+/-SD) for Blacks, Whites, males, and females, respectively. Overall, 132 patients out of 415 (31.8%) were seen in stage I, 61 (14.7%) in stage II, 105 (25.3%) in stage III, 117 (28.2%) in stage IV, while 115 patients (21.7%) had missing stage. RESULTS: There were no significant differences between Blacks and Whites with respect to distributions across stages (P=0.422). Adenocarcinoma was the most common histology in both ethnicities with poor differentiation in 47.4% of Blacks and 57.0% of Whites. Overall, 125 (23.6%) received surgery alone, 54 (10.2%) surgery with chemotherapy, 5 (0.9%) surgery with external radiation therapy, 10 (1.9%) external radiation therapy alone, 68 (12.8%) chemotherapy alone, 58 (10.9%) chemo-external radiation therapy, and 210 (39.6%) no therapy. Overall, only 22 patients survived at 5-yr (4.2%). Median survival was 8.0 months for Blacks (95% CI: 6.4-10.2) and 8.1 months for Whites (95% CI: 7.1-9.7) and there was no significant difference in survival between Blacks and Whites (P=0.971). There were more survivors in females (43/255, 16.9%) than in males (25/275, 9.1%; P=0.009), and females had significantly greater survival times as compared to males (P=0.022). CONCLUSIONS: Pancreatic cancer is a disease of both ethnicities with a slight male predominance among Whites and female predominance among Blacks. We did not find any significant difference in the treatment specific outcome and survival between Blacks and Whites. PMID- 15883476 TI - Management of the mucin filled bile duct. A complication of intraductal papillary mucinous tumor of the pancreas. AB - CONTEXT: Biliary obstruction secondary to intrabiliary mucin is a relatively rare presentation of malignant intraductal pancreatic mucinous tumor. We report a case of unresectable intraductal pancreatic mucinous tumor associated with obstructive jaundice secondary to intrabiliary mucin. This case and a review of the literature, highlight the difficulty in obtaining sustained palliation from jaundice using endoscopically placed biliary stents or percutaneously placed biliary catheters due to rapid occlusion with thick mucin secreted by the tumor. Furthermore, this case differs from that commonly seen in the setting of pancreatic adenocarcinoma, where endoscopic or percutaneous biliary drainage is usually successful at long-term palliation from jaundice. CASE REPORT: Case report We report a case of obstructive jaundice secondary to invasive intraductal pancreatic mucinous tumor associated with dilated bile ducts containing copious amounts of mucin. The diagnosis of intraductal pancreatic mucinous tumor was established based on diagnostic findings on computed tomography scan and endoscopic retrograde cholangiopancreatography. The tumor was unresectable due to vascular invasion. Attempts at endoscopic biliary drainage proved unhelpful with the patient experiencing rapid occlusion of the biliary stents with thick mucinous material leading to recurrent cholangitis. The patient eventually underwent a choledochojejunostomy leading to complete and sustained resolution of the cholestasis. CONCLUSION: If intraductal pancreatic mucinous tumor in association with intrabiliary mucinous obstruction is deemed unresectable, surgical biliary bypass seems to be superior to endoscopic biliary drainage and should be performed on initial presentation. This is due to rapid occlusion of biliary stents with thick mucin leading to frequent stent changes and recurrent cholestasis. PMID- 15883477 TI - Autoimmune pancreatitis associated with retroperitoneal fibrosis. AB - CONTEXT: Autoimmune pancreatitis is sometimes associated with other autoimmune diseases. We have presented two cases of autoimmune pancreatitis with retroperitoneal fibrosis and compared our findings with those found in the literature. CASE 1: A 71-year-old male developed anorexia and weight loss. Abdominal ultrasonography (US) and computed tomography (CT) showed diffuse swelling of the pancreas and the peritoneal soft tissue surrounding the aorta, associated with right hydronephrosis. Endoscopic retrograde pancreatography showed narrowing of the main pancreatic duct. He was diagnosed as having autoimmune pancreatitis associated with retroperitoneal fibrosis and underwent steroid therapy. After 3 weeks, a follow-up CT showed a marked reduction in the size of both the pancreas and retroperitoneal mass. CASE 2: A 62-year-old male was admitted to another hospital complaining of obstructive jaundice. Abdominal CT and US showed swelling of the pancreas. Endoscopic retrograde cholangiopancreatography demonstrated stenosis of the lower bile duct and narrowing of the main pancreatic duct. With the diagnosis of pancreatic head carcinoma, a choledochojejunostomy and a gastrojejunostomy were performed. Histological examination of the biopsy of the pancreatic mass revealed marked fibrosis with lymphoplasmacytic infiltration. One year later, a retroperitoneal mass was detected on follow-up CT. He was treated with prednisolone for two years. Recurrence of retroperitoneal mass with left hydronephrosis occurred 18 months later. There was no sign of recurrence of the autoimmune pancreatitis. He was again treated with prednisolone, and the retroperitoneal mass was gradually reduced. CONCLUSIONS: A total of 7 cases including the present cases have been reported. All were middle-aged males. Steroid therapy was effective for both the pancreatic and the retroperitoneal masses. PMID- 15883478 TI - A case of acute pancreatitis possibly associated with combined salicylate and simvastatin treatment. AB - CONTEXT: Drug-induced acute pancreatitis is a rather rare clinical entity. From time to time, several cases have been reported in which statins or salicylates have been associated with the development of acute pancreatitis. There is only one report which implies the involvement of both drugs in pancreatic inflammation. CASE REPORT: A 58-year-old Caucasian male with a history of coronary heart disease and hypercholesterolemia, under treatment with acetyl salicylate for 6 years and simvastatin for 2 months, presented to the Emergency Department of our hospital with epigastric pain and vomiting of 24-hour duration. The clinical and laboratory investigation led to the diagnosis of acute pancreatitis. Conservative and rich-in-fluid treatment resulted in clinical and laboratory amelioration, and the patient was discharged on day 15, after full restoration of his health. In our patient, all possible common causes of acute pancreatitis were excluded. CONCLUSION: Conclusion It is a rational assumption to connect this case to the co-administration of simvastatin and acetyl-salicylate. However, the pathophysiological mechanism behind the onset of acute pancreatitis due to a statin, or, even more, due to its combination with salicylate, remains vague. PMID- 15883479 TI - IgG4 negative sclerosing cholangitis associated with autoimmune pancreatitis. AB - BACKGROUND: Sclerosing pancreatitis is an autoimmune condition characterized by periductal lymphocytic infiltration on histology, and elevated serum auto antibodies and IgG4. Bile duct involvement is often associated with sclerosing pancreatitis but it is rarely the dominant feature. CASE REPORT: We report a patient with dominant biliary stricture and obstructive jaundice associated with IgG4 negative autoimmune pancreatitis. Due to uncertainties regarding the preoperative diagnosis, the patient underwent bilio-enteric bypass with an intraoperative pancreatic and bile duct biopsy. Post operatively, due to disease recurrence, the patient was started on steroids with consequent remission of the disease. CONCLUSION: Sclerosing pancreatitis may sometimes present with biliary stricture due to bile duct involvement due to the inflammatory process. The condition is often recognized after surgical exploration for a suspected malignancy. Preoperative diagnosis permits treatment with steroids and endoscopic biliary decompression, and avoids unnecessary surgery. PMID- 15883480 TI - Islet isolation and transplantation from an annular pancreas: a case report. AB - CONTEXT: Annular pancreas is an uncommon congenital anomaly formed by a thin band of normal pancreatic tissue encircling the duodenum. CASE REPORT: We report the first case of an islet isolation and transplantation from an annular pancreas. The pancreas together with duodenum was procured from a 32-year-old previously healthy man after diagnosis of brain death. The pancreas including the annular portion was distended well after intraductal collagenase perfusion. A total of 276,064 islet equivalent was recovered and transplanted into a type 1 diabetic patient. CONCLUSIONS: Bearing in mind the shortage of donors, patients with this anomalous condition should not be excluded as potential organ donors. PMID- 15883481 TI - MRCP and ERCP in Lemmel syndrome. PMID- 15883482 TI - Gallbladder motility and acute pancreatitis. PMID- 15883483 TI - JSAR Outstanding Research Award. In vitro growth of mammalian oocytes. AB - The mammalian ovary contains a huge number of small follicles of various sizes, and each follicle encloses a small oocyte. Only a small number of non-growing oocytes (30 microm in the pig and cow) grow to their final size (120 microm), mature, and are ovulated. In vitro growth (IVG) culturing of small oocytes will provide a new source of mature oocytes for livestock production. Using the IVG culture system, non-growing mouse oocytes in primordial follicles grow to their final size and acquire full developmental competence. Among large animals, babies were produced from ovarian oocytes by IVG culture only in the cow. However, the oocytes used were not non-growing ones but at the mid-growth stage (90-99 microm in diameter) in early antral follicles. Xenotransplantation of the follicles at an early stage to immuno-deficient mice is a substitute for an effective long term IVG culture of much smaller oocytes. IVG and xenotransplantation of small oocytes at a specific size will provide a new understanding of the mechanisms regulating oogenesis and folliculogenesis in the complex mammalian ovary. PMID- 15883484 TI - JSAR Innovative Technology Award. Development of ovulation synchronization and fixed time artificial insemination in dairy cows. AB - Recently, reproductive management has become more difficult as a result of increased herd size. Problems with missing estrous signs and decrease in conception rate by artificial insemination (AI) performed at wrong timing have caused low AI conception rates. In 1995, ovulation synchronization and fixed-time AI (Ovsynch/TAI) was developed in the USA as a new reproductive technology, which was accepted as an useful reproductive management tool in many countries. However, no information on the use of Ovsynch/TAI was available in Japan. It was, therefore, warranted to show the ovulation rate and conception rate after Ovsynch/TAI using gonadotropin releasing hormone analogue (GnRH-A, fertirelin acetate) and prostaglandin F2alpha (PGF2alpha)-THAM, both were commercially available in this country. The conception rate after Ovsynch/TAI has been known to vary among different herds and individuals. Investigation and analysis of factors affecting the conception rate was also warranted to improve the conception rate. A series of experiments were carried out to establish Ovsynch/TAI using domestically produced GnRH-A and PGF2alpha and to study factors affecting conception rate after Ovsynch protocol. Ovsynch using 100 microg GnRH-A and 25 mg PGF2alpha were observed using ultrasonography. As a result, a high synchronization rate of ovulation at 16 to 20 h after the second GnRH injection was confirmed. The conception rate after Ovsynch/TAI was compared in 87 cows with the conception rate after AI at estrus induced by PGF2alpha (139 cows). Conception rate after Ovsynch/TAI was higher than the figure after AI at induced estrus (59.1% vs 20.9%, P<0.05). The dose of GnRH-A was also studied and a practical dose of GnRH-A was found to be 50 microg per cow. To clarify some factors affecting the conception rate after Ovsynch/TAI, 1,558 cows were investigated for the state of their ovaries, days after calving, parity, season, ovarian cyclicity postpartum and nutritional state at the day of Ovsynch. The overall conception rate after Ovsynch/TAI was 51.5%. Fifty-six cows (3.6%) showed estrus at 6 to 7 d after the first injection of GnRH-A. The conception rate after Ovsynch/TAI was low in cows that were 40 to 60 d postpartum, those in their 5th lactation or more, those bred in July to August, and those recovering ovarian cyclicity later than 56 d postpartum. The conception rate after Ovsynch/TAI was high in cows in which body condition score (BCS) was 3.75 at dry period and 3.0 at the day of Ovsynch. In conclusion, Ovsynch/TAI is an effective tool for the reproductive management of dairy cows. A steady and sufficient conception rate after Ovsynch/TAI could be expected by taking the factors affecting the conception rate into the consideration. PMID- 15883485 TI - Permeability of mouse oocytes and embryos at various developmental stages to five cryoprotectants. AB - To assess the permeability of mouse oocytes and embryos, matured oocytes and embryos at various stages of development were placed in five cryoprotectant solutions at 25 C for 25 min. From the cross-sectional areas of the oocytes/embryos, the relative change in volume was analyzed. In oocytes, shrinkage was least extensive and recovery was quickest in the propylene glycol solution, showing that propylene glycol permeates the oocytes most rapidly. Dimethyl sulfoxide, acetamide, and ethylene glycol permeated the oocytes slightly more slowly than propylene glycol. The oocytes in glycerol shrunk extensively and then expanded marginally, indicating slow permeation. The volume changes of 1 cell and 2-cell embryos were similar to those of oocytes, showing little change in permeability. In 8-cell embryos, the volume recovered much faster than in the earlier stages especially in glycerol and acetamide. In morulae, the volume recovery was much faster in glycerol and in ethylene glycol; in ethylene glycol, the extent of shrinkage was small and the recovery was fast, indicating an extremely rapid permeation. Although the permeability of oocytes/embryos generally increased as embryo development proceeded, the degree of increase varied greatly among the cryoprotectants. Interestingly, the volume change in propylene glycol was virtually unaffected by the stage of development. Such information will be valuable for determining a suitable protocol for the cryopreservation of oocytes/embryos at different stages of development. PMID- 15883486 TI - Conflict of estrogenic activity by various phthalates between in vitro and in vivo models related to the expression of Calbindin-D9k. AB - Phthalates are suspected to disrupt the endocrine system, especially through estrogenic effects. In the present study, we investigated the effects of various phthalates and compared them with those of estrogenic compounds that disrupt the female reproductive system. To assess the effects of these phthalates, alteration of the Calbindin-D9k (CaBP-9k) gene was measured as a biomarker because rat CaBP 9k gene carries an estrogen response element (ERE) which is involved in estrogen responsiveness of the gene during the estrous cycle. In this study, phthalates were tested for estrogenic properties in in vitro and in vivo models. First, the E-Screen assay was used to measure the proliferation of MCF-7 cells, a human breast cancer cell line. Treatments with 17beta-estradiol (E2; 9-fold) and 17alpha-estradiol (EE; 9-fold) induced MCF-7 cell proliferation at concentrations of 10(-9) M. Phthalates induced an increase in MCF-7 proliferation at concentration of 10(-6) M up to 10(-4) M. Nbutyl benzyl phthalate (BBP; 6-fold vs. vehicle), dicyclohexyl phthalate (DCHP; 8-fold), 2-ethylhexyl phthalate (DEHP; 6-fold) and di-n-butyl phthalate (DBP; 7-fold) at the concentration of 10( 4) M induced in an increase in MCF-7 proliferation after 6 d of treatment compared to vehicle. However, significant increase in MCF-7 proliferation was induced by diethyl phthalate (DEP). Second, we investigated the expression of CaBP-9k in the uterus of immature rats after oral treatment with BBP, DCHP, DEHP, DBP or DBP (600 mg/kg per day) in this in vivo model, because the immature rat model is highly sensitive to exposure to estrogenic chemicals. None of the phthalates induced the expression of CaBP-9k mRNA and its protein in the neonatal uterus as analysed by Northern and Western blot analyses, respectively. Although phthalates induced an increase in MCF-7 cell proliferation by an estrogenic effect, they could not induce CaBP-9k expression in the in vivo system, suggesting that the assays of estrogenic effects of various phthalates conducted in vitro and in vivo expression of CaBP-9k may produce conflicting results. PMID- 15883487 TI - [Functional foods or nutritional-dose supplementations: what does the future hold for micronutrition in ophtalmology?]. PMID- 15883488 TI - [Comparison of three antiallergic eyedrops in a specific conjunctival provocation test: mequitazine, levocabastine and dexamethasone]. AB - PURPOSE: In order to assess the efficacy of an antihistaminic eye drop containing 0.05% mequitazine in the prevention of allergy induced by a conjunctival provocation test, a double-masked, randomized, intraindividual study compared this eye drop to 0.05% levocabastine and 0.1% dexamethasone eye drops in 24 subjects allergic to grass pollen. METHOD: During the first phase of treatment, randomized subjects received one drop of dexamethasone in one eye and one drop of either mequitazine or levocabastine in the fellow eye. During the second phase of treatment, they were given one drop of dexamethasone in the same eye as previously, and one drop of the treatment that had not been given during the first phase (levocabastine or mequitazine) in the fellow eye. Fifteen minutes after each instillation phase, a conjunctival provocation test was performed. Hyperemia, itching, tearing, chemosis and palpebral edema were the five signs or symptoms taken into account to assess the treatment efficacy. Their intensity was evaluated 3, 5 and 10 min after the conjunctival provocation test. The primary efficacy criterion was the global score obtained by measuring hyperemia and itching intensity. RESULTS: The score was reduced significantly more (p < 0.0001) for the eyes treated with mequitazine or levocabastine than for those treated with dexamethasone at all evaluation times. The difference was also significant for hyperemia (p < 0.001), itching (p < 0.001), and tearing (p < 0.05). The tolerability of the three eyedrops was satisfactory. CONCLUSION: Mequitazine and levocabastine were safe and significantly more effective than dexamethasone in preventing the allergic response induced by a conjunctival provocation test when they were instilled 15 min before contact with the allergen. PMID- 15883489 TI - [Cornea donation: improving hospital coordination]. AB - PURPOSE: To evaluate the impact of procedures developed to increase cornea donation in a general hospital. Comparative study before and after intervention. Since february 2002 all cases of intra-hospital death were considered by the hospital coordinator. A written selection protocol for potential donors was adopted and telephone interviews to obtain family consent were conducted using a standardized procedure. METHODS: The number of deaths in the institution, deaths considered by the hospital coordinator, clinical contraindications to harvesting, potential donors, face-to-face interview and telephone contact with families, the donation acceptance rate during telephone and face-to-face interviews, and the number of donors and corneas actually collected were recorded before and after intervention. The data were analyzed during two consecutive periods: Period 1, february 2001 to january 2002 (before protocol), and period 2, february 2002 to january 2003 (after protocol). RESULTS: The number of deaths in the institution was similar for the two periods. The increase in deaths considered by the hospital coordinator and the decrease in contraindications resulted from increasing the pool of potential donors from 188 to 401 (p < 0.001). The number of telephone interviews increased from 33 to 274. The rate of family consent given during face-to-face interviews was similar, 61% versus 72% (p = 0.1), but acceptance during telephone interviews increased from 15% to 60% (p < 0.001) and 138 corneas were collected in period 1 and 443 in period 2. The rate of cornea donors increased from 6% to 20% of the hospital's deaths. CONCLUSION: The consideration of all hospital deaths, the use of a written protocol of contraindications for cornea donation and the wide use of telephone interviews to obtain family consent led to a dramatic increase in cornea donation. PMID- 15883490 TI - [Multifocal phakic intraocular lens implant to correct presbyopia]. AB - INTRODUCTION: Presbyopic surgery is considered as the new frontier in refractive surgery. Different solutions are proposed: myopization of one eye, insertion of an accommodative crystalline lens, scleral surgery, the effects of which are still unknown, and finally multifocal phakic implants. We therefore decided to undertake a prospective study under the Huriet law to determine its efficacy and specify the conditions required for an anterior chamber multifocal phakic implant. MATERIAL AND METHOD: Fifty-five eyes of 33 patients received an anterior chamber foldable multifocal phakic implant. Twenty-one females and 12 males underwent surgery. Initial refraction was between -5D and +5D. The implant's single addition was +2.50. Recuperating a distant uncorrected visual acuity of 0.6 or better and near uncorrected vision of Parinaud 3 or better can be considered a very good postoperative result. RESULTS: Average follow-up was 42.6+/-18 weeks. Mean postoperative refraction was -0.12+/-0.51 D. Mean postoperative uncorrected visual acuity was 0.78+/-0.20. Postoperative uncorrected visual acuity was Parinaud 2.3+/-0.6. Eighty-four percent of eyes operated on recuperated 0.6 or better without correction and Parinaud 3 or better without correction. Lenses in four eyes were explanted for different reasons, essentially optical, and no severe anatomical complications were observed. CONCLUSIONS: Placing an anterior chamber multifocal phakic implant to correct presbyopia is an effective technique with good predictability and has the advantage of being reversible in case of intolerance, optical parasite effects or undesired complications. Considering the particularity of this surgery, it is imperative to respect very strict inclusion criteria: anterior chamber depth equal to or above 3.1 mm, open angle, endothelial cell count equal to or above 2000 cells/mm2, absence of an incipient cataract or the slightest evidence of macular alteration. PMID- 15883491 TI - [Urgent penetrating keratoplasty in perforated infectious corneal ulcers]. AB - PURPOSE: To evaluate anatomical and functional prognosis of urgent penetrating keratoplasty in perforated infectious corneal ulcers. METHODS: Seven eyes of seven patients had an urgent penetrating keratoplasty for perforated infectious corneal ulcers. Anatomical success was defined by eradication of infection and preservation of eye from enucleation and phthisis bulbi. Corneal graft survival was defined by the presence of a clear graft. RESULTS: The patients'mean age was 40.5 years. Four eyes had bacterial corneal ulcer and three eyes had herpetic corneal ulcer. Anatomical success was obtained in six eyes. The graft remained clear in two eyes. Graft rejection was noted in four eyes after a mean period of 4.5 months. Postoperative complications were ocular inflammation (one eye), ocular hypertension (three eyes), cataract (one eye), peripheral anterior synechiae (one1 eye), graft ectasia (one eye), bacterial infection (one eye) and recurrent herpetic keratitis (one eye). The mean follow-up period was 22 months, ranging from 9 to 32 months. CONCLUSION: Urgent penetrating keratoplasty can preserve eye integrity and eradication of the infectious process in a large part of perforated bacterial and herpetic corneal ulcers. Visual rehabilitation is often a secondary objective. Adapted antimicrobial treatment reduces graft reinfection and steroid treatment reduces the frequency of some complications, especially graft rejection. PMID- 15883492 TI - [Tissular and mechanical effects observed with an experimental femtosecond laser microkeratome for corneal refractive surgery]. AB - PURPOSE: Despite progress in mechanical microkeratomes used in refractive surgery, mechanical complications during cutting of the cornea still occur. Cutting by laser could reduce these complications and to date, the femtosecond laser is the only potential candidate for this purpose. Our study reports preliminary results with a femtosecond microkeratome for cutting porcine corneas ex vivo. METHODS: We first examined the fundamental principles of the interaction between the femtosecond laser and the corneal stroma, including the volume of tissue lesions, the laser breakdown threshold of the stroma and the laser ablation selectivity. We then analyzed the quality of cutting corneal flaps with the laser, focusing on collateral tissue effects and the roughness of the interfaces observed both histologically and with scanning electron microscopy. RESULTS: The photoablative and photodisruptive effects were very similar with the femtosecond laser. This characteristic is specific to ultrashort impulsion photodisruptor lasers and allows for a very precise surgical procedure. The laser induced breakdown threshold of porcine corneal stroma was found to be 0.55 J/cm2. Collateral tissue lesions were on the submicrometer level. The roughness of the stromal bed was optimal for postage stamp cutting, providing very many contiguous points of impact which were as spherical as possible. CONCLUSION: Corneal photodisruption with a femtosecond laser is reproducible and extremely accurate. The optomechanical parameters involved with this technique require great technological skill and should be placed in experienced hands. PMID- 15883493 TI - [Bimatoprost in the treatment of ocular hypertension and chronic glaucoma]. AB - PURPOSE: The aim of this study was to evaluate the efficacy and the safety of bimatoprost in an outpatient glaucoma practice and to correlate the responsiveness to this treatment with the central corneal thickness. MATERIALS AND METHODS: Our retrospective analysis included 55 consecutive patients (mean age, 66 years). Bimatoprost was administered in monotherapy in 32 patients and in combined treatment in 23. Mean follow-up was 5.5 months. In bilateral treatments (33/55 patients), only one eye (with the more severe defect and/or the higher IOP) was included in the analysis. The patients were considered as responders to bimatoprost when the observed reduction of IOP was > or = 20% and/or at least 3 mmHg compared with the pretreatment IOP. The mean central corneal thickness (CCT) was extrapolated from five consecutive measurements with the ultrasonic pachymeter Pachette. RESULTS: Overall, the mean IOP was reduced from a pretreatment value of 21.1 mmHg to 17.3 mmHg at the last visit (mean IOP decrease, 3.6 mmHg, or 17%) (p < 0.05). Except for four patients (7.3%) who discontinued bimatoprost secondary to local or systemic adverse effects, ocular tolerance of bimatoprost was excellent in 62%. Moderate conjunctival hyperemia was present in 18%. The mean IOP reduction was 19% in monotherapy and 15% in combined treatments. Concomitantly, the percentage of responders was slightly higher in patients only receiving bimatoprost than in patients receiving bimatoprost associated with other medication (s). In monotherapy, bimatoprost induced a further IOP decrease of 12% compared with a previous association of two medications that did not include a prostaglandin (10 patients). In the 20 patients in whom bimatoprost had replaced another prostaglandin, a further mean IOP reduction of 11% was observed. The frequency of distribution of the responders to bimatoprost was not correlated with CCT (chi2, p > 0.05). CONCLUSIONS: Considering the limits of this study, our results suggested that bimatoprost was effective and well tolerated in most patients. The decrease in IOP and responsiveness to treatment appeared to be slightly higher in monotherapy than in combined treatments, equivalent to a combination of two medications without prostaglandin and equivalent to or slightly higher than other prostaglandins. The degree of responsiveness did not seem to be correlated with CCT. PMID- 15883494 TI - [Role of trypan blue in epiretinal membrane surgery]. AB - INTRODUCTION: Idiopathic epiretinal membrane (ERM) results from detachment of the posterior hyaloid. Vitrectomy and peeling are the treatment of choice. Trypan blue 0.15% (TB) stains the epiretinal membrane and the internal limiting membrane (ILM), facilitating surgery and thus allowing selective and complete removal with less retinal damage. We conducted a study evaluating the advantages of selective staining associated with surgery, the incidence of postoperative damage, and the influence on final visual acuity. MATERIAL AND METHODS: Fourteen patients were operated on by the same surgeon, who carried out a complete vitrectomy, then fluid-air exchange for dye injection of undiluted 0.2 ml of Trypan blue 0.15% in front of the membrane, which was then easily distinguished from the area under the retina. Preoperative visual acuity, postoperative damage, as well as eye status at the end of follow-up are discussed. Transmission electron microscopy was used to determine if epiretinal membrane removal was complete and selective. Angiography was systematically used to study the state of retina before and after surgery. RESULTS: Preliminary results show that trypan blue is useful and facilitates complete removal of ERM and ILM. It may reduce long-term complications and recurrent membrane formation. Trypan blue is a staining agent that is quick and easy to use. It presents no short-term toxic effects. Functional results in 6 months show improvement of visual acuity and disappearance of metamorphopsia. CONCLUSION: Idiopathic epiretinal membrane surgery with tryptan blue staining is a better alternative than indocyanine green staining. PMID- 15883495 TI - [Quantitative and spatial analysis in image processing: study of drusen distribution from the foveal center in age-related macular degeneration]. AB - Angiofluorograms were obtained from 58 patients presenting posterior pole drusen due to age-related macular degeneration. These frames underwent image processing by morphological mathematics algorithms. The detection sensitivity of this algorithm is 78%. We present a quantitative and spatial analysis of drusen distribution from the foveal center in the form of a graph. This technique may allow for multi-date comparisons, by using a graphic representation of this risk factor for age-related macular degeneration. PMID- 15883496 TI - [Refractive phakic IOLs: three different models and contact with the crystalline lens. An AC-OCT study]. AB - Three patients received three different models of phakic implants. With the usual slit-lamp examination method, we were not able to establish whether there was contact between the implants and the natural crystalline lens. Using the anterior chamber optical coherence tomography scanner (AC-OCT), we were able to demonstrate that there was direct contact between the natural crystalline lens and the three different phakic implants. A dynamic study of these contacts was carried out during accommodation. These observations show that examination of the anterior segment with the optical coherence tomography scanner is essential in refractive surgery. PMID- 15883497 TI - [Evaluation of capsular block syndrome with an anterior segment OCT]. AB - A year after routine phacoemulsification and insertion of a soft posterior chamber IOL, a 59-year-old woman developed progressive moderate myopia. A syndrome of late capsular block was diagnosed following biomicroscopic examination, which was treated by Nd YAG laser posterior capsulotomy. The myopia disappeared immediately. This observation was illustrated with optical coherence tomography (OCT) developed for the anterior segment. After capsulotomy, the IOL moved backwards by 448 microm, which corresponds exactly to a--1D induced myopia. PMID- 15883498 TI - [Hypothesis on the role of nutritional factors in ocular hypertension and glaucoma]. AB - Recent notions in connection with oxidative stress and the fat balance of long chain polyunsaturated fatty acids (PUFA) families have brought new insight to a probable role of nutritional factors in glaucoma and intraocular hypertony. The modifications of the extracellular matrix of the trabecula could be influenced by oxidative stress. On the one hand, collagen apoptosis and remodeling (associated with an increase in intraocular pressure) are mainly influenced by hydrosoluble antioxidants such as glutathione. On the other hand, elastin apoptosis and remodeling (correlated with the occurrence of optic atrophy) are particularly influenced by liposoluble antioxidants such as vitamin E. In addition, the dietary ratio of omega3/omega6PUFA intake could influence the balance of intraocular pressure. Omega-3 PUFA could influence cyclooxygenase competition. A diet with increased omega-3 and decreased omega-6 could thus favor an increase in intraocular pressure reducing synthesis of PG-F2, leading to a decrease in uveoscleral outflow. The true importance of these factors has not yet been solidly determined and studies are in progress to clarify the real implication of these nutritional factors. PMID- 15883499 TI - [Circadian variations in intraocular pressure and their clinical implications]. AB - Intraocular pressure follows a circadian rhythm like many of the body's parameters. These fluctuations in intraocular pressure were recently identified as a risk factor for visual field loss, in addition to intraocular pressure itself. Furthermore, the pressure curve seems to have a different profile in glaucomatous patients: the peak occurs later compared with normal subjects. Better knowledge of pressure fluctuations and their physiological mechanisms is essential for optimal management of glaucomatous patients not only to interpret the single ocular tension recordings, but also to use the available treatments rationally, both drugs and surgery. The authors review different studies that have contributed to the current knowledge on the circadian rhythm of intraocular pressure, aquous humor flow regulation, and the effects of topical hypotensive drugs on the pressure curve. PMID- 15883500 TI - [PPAR gamma: a novel pharmacological target against retinal and choroidal neovascularization]. AB - PPARg (peroxisome proliferator-activated receptor gamma) is a nuclear receptor that regulates the transcription of numerous genes involved in the differentiation, proliferation and apoptosis of various cell types. It was initially discovered in adipocytes as a differentiation agent, then was characterized in vascular endothelium and recently in choroidal and retinal endothelial cells. Agonists that bind to PPARgamma and stimulate its transcriptional activity are endogenous lipids such as lysophosphatidic acid and 15-d-PGJ2 as well as the synthetic pharmacological compounds, thiazolidinediones, used for treating type 2 diabetes. These ligands prevent choroidal and retinal neovascularization in several experimental animal models, notably through the inhibition of vascular endothelial growth factor (VEGF) receptor expression. Because of the high affinity and the low molecular weight of agonists, suitable for good bioavailability, PPARgamma could potentially be a novel pharmacological target of angiostatic agents, particularly useful to treat age-related macular degeneration and diabetic retinopathy. PMID- 15883501 TI - [Nodular melanoma on primary acquired conjunctival melanosis]. AB - A 74-year-old woman consulted for bloody tears. The etiology was a large conjunctival nodular melanoma hidden in the left superior fornix that had developed quietly on an unknown primary acquired melanosis. In this report the clinical and histological features as well as the treatment are presented. A decisional tree summarizes the treatment for conjunctival melanosis. PMID- 15883502 TI - [The development of ambulatory surgery: how far have we come?]. PMID- 15883503 TI - [Splenic cysts and tumors: diagnosis and management]. AB - Cysts and tumors of the spleen are rare and are often discovered fortuitously. They are most often asymptomatic but may present with abdominal pain in the left upper quadrant. Splenic cysts are far more common than solid lesions; true cysts must be differentiated from pseudocysts of the pancreas and from cystic degeneration following splenic contusion or infarction. Cysts may be congenital (epidermoid cysts), infectious (abscess or hydatid cyst), or neoplastic (lymphangioma or angioma with tumor necrosis). Diagnosis can usually be established with the clinical context and imagery (ultrasound, CT, MRI). Surgery should be avoided for angiomas and pseudocysts. Spleen-conserving surgery is indicated for large symptomatic epidermoid cysts. Splenectomy is often required for hydatid cysts and tumors. Of the solid tumors, hemangiomas and lymphangiomas often have a characteristic fleshy appearance. For other solid tumors, whether benign or malignant, imaging may give some clues to the diagnosis, but diagnostic certitude often requires pathologic examination of the piece. Needle biopsy is contraindicated because of the risk of bleeding. Resection should be as limited as possible in order to avoid the risks of total splenectomy (overwhelming sepsis, thrombosis). PMID- 15883504 TI - [Perioperative care in digestive surgery]. PMID- 15883505 TI - [Colectomy for cancer involving the apex of the sigmoid loop-open surgical approach]. PMID- 15883506 TI - [Laparoscopic appendectomy]. PMID- 15883507 TI - [Postero-lateral thoracotomy]. PMID- 15883508 TI - [Internal hernia after laparascopic left colectomy; a simple method of prevention]. PMID- 15883509 TI - [Internal hernia of the broad ligament]. PMID- 15883510 TI - [A complicated case of Crohn's disease]. PMID- 15883511 TI - [Ars longa, vita brevis]. PMID- 15883513 TI - [Did you say "evidence-based medicine?"]. PMID- 15883514 TI - [Cecal carcinoma presenting as an ileoocolic intussusception]. PMID- 15883515 TI - Image fusion: a new diagnostic and therapeutic tool. PMID- 15883516 TI - Occult inflammatory breast cancer: review of clinical, mammographic, US and pathologic signs. AB - PURPOSE: To examine the clinical, radiologic and pathologic findings of occult inflammatory breast cancer (OIBC) in order to identify features useful for diagnosis. MATERIALS AND METHODS: We retrospectively reviewed the records of 19 women with OIBC observed at our Department between 1992 and 2001. We analysed the clinical history, mammographic, ultrasonographic, and pathologic findings and investigated overall survival (OS), prognostic variables and radio-pathologic correlations. RESULTS: The most common mammographic findings were: diffusely increased density (52.63%), trabecular thickening (42.1%), mass (36.84%). The most common US findings were axillary lymphadenopathy (68.75%), skin thickening (43.75%) and mass (56.25%). At least one inflammatory sign was found in 14 women (74%) at mammography (subcutaneous thickening, trabecular thickening, diffuse increase in density) or at US (subcutaneous thickening, diffuse increase in echogenicity due to oedema, lymph vessel dilatation). Estrogen receptors (ER) were present in 63.2% and Progesterone receptors (PgR) in 36.8%. Significant prognostic variables were ER and Ki 67. CONCLUSIONS: The typical radiological pattern of clinical inflammatory breast carcinoma is less frequently present in OIBC; nevertheless the radiologist must pay attention because frequently OIBC presents just one radiological sign and this should be enough for a diagnostic suspicion. Moreover, the absence of clinical and radiological inflammatory signs does not exclude inflammatory breast cancer because OIBC can manifest at imaging as a mass or isolated calcifications. ER and PgR are positive in a high percentage of patients and confirm that OIBC has a better prognosis than clinical inflammatory breast cancer. PMID- 15883517 TI - Role of multimedial diagnosis of breast cancer in women below 36 year of age. AB - PURPOSE: To evaluate the sensitivity for breast cancer of different diagnostic tests performed in a consecutive series of women aged under 36 years. MATERIALS AND METHODS: The study analyses 155 women with breast cancer incident in the Tuscany Cancer Registry from 1985 to 2000. The sensitivity of each method was evaluated in relation to the most recent test performed during the year before diagnosis and to different variables, such as tumour size and calendar period. RESULTS: Sensitivity was 70.3 % for physical examination, 76.0% for mammography, 69.1% for ultrasonography and 80.6% for cytology (86.2% if inadequate samples are excluded). Sensitivity was associated to pT category for physical examination (pT1=60.6%; pT2-4=86.4%; chi squared=10.2, p= 0.001) and for ultrasonography (pT1=61.9%; pT2-4=92.0%; chi squared=5.7, p=0.01) and to breast radiological density for mammography (50-100% density=75.5%; 0-50% density=91.3%; chi squared=1.85, p<0.17). Among 112 cases examined with palpation, mammography and cytology, these methods had a sensitivity of 78.5%, 75.8% and 91.9%, respectively, and an overall sensitivity of 98.2%. Among 65 cases examined with palpation, ultrasonography and cytology, these methods had a sensitivity of 67.6%, 70.7% and 92.3%, respectively, and an overall sensitivity of 95.3%. Among 123 cases examined with palpation, imaging (mammography and/or ultrasonography) and cytology, these methods had a sensitivity of 76.4%, 78.0% and 90.2%, respectively, and an overall sensitivity of 96.7%. Cancer was suspected by one method only in 2.4% of cases with palpation, 1.6% with imaging and 7.3% with cytology. Comparison between mammography and ultrasonography was possible in 58 cases undergoing both tests: sensitivity was 79.3% for ultrasonography and 74.1% for mammography, respectively chi squared=0.1, p=0.66). If sensitivity had been determined according to the test performed immediately before diagnosis it would have been higher, that is 74.4% for palpation, 76.8% for mammography, 73.5% for ultrasonography and 90.9% for cytology (97.7% if inadequate samples were excluded), and overall sensitivity would be 96.1%. CONCLUSIONS: Differential diagnosis of breast cancer in women aged under 36 years is difficult, sensitivity being lower as compared to older women, with the only exception of cytology. The association of multiple tests (diagnostic imaging and cytology) is crucial to achieve an acceptable false negative rate. In particular, the extensive use of cytology in the presence of an even minimal doubt at palpation and/or imaging is strongly recommendable. Sensitivity must be calculated on the basis of an independent pathology registry, and false negatives must be defined within a given period after the test. Analysis limited to tests performed immediately before diagnosis leads to a biased overestimation of sensitivity. PMID- 15883518 TI - Role of axillary lymph node ultrasound and large core biopsy in the preoperative assessment of patients selected for sentinel node biopsy. AB - PURPOSE: The aim of this study was to evaluate the diagnostic accuracy of axillary lymph node sonography, if necessary in combination with US-guided large core biopsy, in the preoperative evaluation of breast cancer patients scheduled for quadrantectomy and sentinel lymph node excision. MATERIALS AND METHODS: From July 2001 to December 2002, we evaluated 117 breast cancer patients with ultrasound and, where indicated, FNAB. Breast lesions had diameters between 4 and 26 mm (mean diameter 11 mm). Fifteen (13%) of the 117 patients were excluded from the series as they did not fulfil the criteria for preliminary sonography of the axilla: in 9 patients fewer than 4 nodes were detected and in 6 patients the breast lesions were intraoperatively found to be benign. Eleven patients (10.7 %) with sonographically suspicious axillary nodes were sampled by US-guided core biopsy using a 14 or 16 Gauge Tru-Cut needle. RESULTS: The ultrasound study aims to evaluate the dimensions and morphology of the breast lesion as well as detect and assess at least 4 axillary nodes. These were evaluated for hilar and cortical thickening and ratio between the sinus diameter and the total longitudinal diameter. Lymph nodes with hilar diameters equal to or greater than 50% of the longitudinal diameter were considered normal. Of the 102 patients evaluated, 77 (75.7%) had normal axillary nodes according to the US criteria adopted. Negativity was confirmed by histology in 56 cases (72.7%, true negative); 21 (27.3%, false negative) were found to be positive, in contrast with the sonographic appearance. The false negative cases were due to lymph node micrometastasis which probably did not cause morphologic alterations perceptible at ultrasound. The remaining 25 patients (24.5%) had axillary lymph nodes classified as suspicious. In 13 cases of (52%, true positive) there was agreement with histology, whereas in 12 cases (48%, false positive) the US suspicion was not confirmed at surgery. The most important sonographic alteration was the gradual reduction in hilar echogenicity (seen in 100% metastatic nodes); conversely, hilar denting or irregularities, as well as dimensional criteria, proved to be poorly specific. CONCLUSIONS: The sonographic assessment of axillary and internal mammary chain nodes, possibly in addition to core biopsy, improves the preoperative evaluation of breast cancer patients scheduled for conservative surgery of the breast (quadrantectomy) and the axilla (sentinel node biopsy). US findings suspicious for metastatic involvement of axillary lymph nodes should be considered as an exclusion criterion for sentinel node biopsy. A more widespread use of axillary node biopsy and an accurate sonographic evaluation of the excised lymph node increase the specificity of the procedure, allowing a better correlation between sonographic findings and definitive histology. PMID- 15883519 TI - Evidence Based Medicine (EBM) and Evidence Based Radiology (EBR) in the follow-up of the patients after surgery for lung and colon-rectal carcinoma. AB - PURPOSE: A) To define the role of diagnostic imaging modalities in the follow-up of patients after surgery for solid cancer, using an Evidence Based Medicine (EBM) approach; B) to assess the possible discrepancies between the theoretical model and the clinical protocols currently used for the follow-up of treated patients; c) to compare the real costs of the radiological examinations performed in a group of cancer patients followed up after surgery and the theoretical costs that would have been incurred had the patients been followed up according to the theoretical (evidence-based) follow-up programme. MATERIALS AND METHODS: We searched traditional and secondary databases for research papers and guidelines by international scientific societies published in the last 10 years and concerning the clinical impact of follow-up programs in patients operated on for colorectal and lung carcinoma. The papers were selected based on level of evidence using the systematic review approach of EBM. In each paper selected, we considered the overall survival and disease-free survival, quality of life, side and toxic effects of therapy, costs and psychological aspects to formulate a judgement on the usefulness of the radiological tests. Subsequently, the clinical and imaging follow-up of 40 patients who had undergone surgical resection for colorectal cancer (20 patients) and lung cancer (20 patients) between 1998 and 2004 were retrospectively reviewed, and the costs of the follow-up programs for the two groups were analysed and compared with those of the theoretical evidence based programmes. RESULTS: Of the 41 papers selected after systematic review only nine datasets were considered for our final analysis. The majority of papers (7 out of 9) and all the guidelines published by international Scientific Societies agreed on the poor value of closed imaging in the follow-up of patients who have undergone surgery for colorectal and lung cancer. A significant difference was found between the real costs of the follow-up programmes implemented in the 40 patients considered and the theoretical costs derived from the guideline recommendations (an excess of 99.06% for lung cancer, and 93.6% for colon cancer). CONCLUSIONS: Our findings can serve as a basis to start a discussion within the scientific community about the role of radiological follow-up in cancer patients with the aim of defining a more rational use of resources. PMID- 15883520 TI - Characterization of focal liver lesions with a new ultrasound contrast agent using continuous low acoustic power imaging: comparison with contrast enhanced spiral CT. AB - PURPOSE: To evaluate the concordance of the enhancement patterns of a new ultrasound contrast agent (SonoVue) with those obtained with dual-phase contrast enhanced spiral CT (CE-CT) in the characterization of focal liver lesions (FLLs). MATERIALS AND METHODS: Sixty-two patients with focal liver lesions discovered at ultrasound and also studied with CECT underwent contrast-enhanced ultrasound using continuous low acoustic power imaging after receiving a 2.4 ml bolus of the new US contrast agent SonoVue, consisting of a dispersion of sulphur hexafluoride microbubbles. The examinations were made using ATL HDI-5000, Acuson SEQUOIA and Aloka 5500 Prosound ultrasound systems with 5.2 MHz curved-array probes. The concordance between US and CE-CT images was evaluated on site by two radiologists blinded to CT RESULTS: The FLLs were assessed in the arterial (20 s after CM injection), portal (after 45-60 s) and late (after 120 s) phases for: 1) presence/absence of enhancement 2) distribution of enhancement (homogenous or target distribution, centripetal or centrifugal flow, and other), 3) qualitative enhancement pattern (hyperechoic, hypoechoic, or isoechoic) versus normal liver parenchyma. RESULTS: The concordance between SonoVue-enhanced US and CE-CT was 85%. Moreover during portal venous phase with CEUS it was possible to differentiate between malignancy or benignity of 91% of lesions. CONCLUSIONS: The preliminary data obtained in this study suggest that continuous low acoustic power imaging and contrast-enhanced US show similar results to CT in contrast distribution and contrast enhancement patterns. PMID- 15883521 TI - Ogilvie's syndrome (acute colonic pseudo-obstruction): review of the literature and report of 6 additional cases. AB - PURPOSE: Ogilvie's syndrome is defined as an acute pseudo-obstruction of the colon, characterized by the signs, symptoms and radiological pattern of a large bowel obstruction, but without a detectable organic cause. The aetiology of Ogilvie's syndrome appears to be multifactorial, with a series of possibly interacting pathogenic noxae all resulting in colon inactivity. Our study reports on six cases of Ogilvie's syndrome diagnosed and treated between 1997 and 2002. MATERIALS AND METHODS: From October 1997 to September 2002 we studied six patients affected by pseudo-obstruction of the colon. The pseudo-obstruction was recurrent in two cases. Acute dilatation of the colon without radiologically detectable organic obstruction was the inclusion criterion for the study. RESULTS: Plain abdominal radiography revealed colon dilatation that extended to the splenic flexure in three patients, to the hepatic flexure in two patients, and confined to the transverse colon in one patient. None of the patients showed air-fluid levels of the small intestine. CONCLUSIONS: The most relevant clinical finding in Ogilvie's syndrome is abdominal distension, which arises suddenly, has a progressive course and reaches massive levels. The first-line diagnostic investigation is plain abdominal radiography which shows extreme colon dilation without air-fluid levels of the small intestine. In three of our patients, conservative therapy alone was able to restore normal conditions within five days; two patients required decompressive colonoscopy, and one patient died from cardio-circulatory arrest after 48 hours. PMID- 15883522 TI - Technical analysis of volume-rendering algorithms: application in low-contrast structures using liver vascularisation as a model. AB - AIM: To assess the influence of pre-set volume-rendering opacity curves (OC) on image quality and to identify which absolute parameters (density of aorta, hepatic parenchyma and portal vein) affect visualization of portal vascular structures (low-contrast structures). MATERIALS AND METHODS: Twenty-two patients underwent a dual-phase spiral CT with the following parameters: collimation 3 mm, pitch 2, increment 1 mm. Three scans were performed: one without contrast medium and the latter two after the injection of contrast material (conventionally identified as ''arterial'' and ''portal''). The images were sent to a workstation running on an NT platform equipped with post-processing software allowing three dimensional (3D) reconstructions to generate volume-rendered images of the vascular supply to the liver. Correlation between the absolute values of aorta, liver and portal vein density, OC parameters, and image quality were assessed. RESULTS: 3D images generated using pre-set OC obtained a much lower overall quality score than those produced with OC set by the operator. High contrast between the liver and the portal vein, for example during the portal vascular phase, allows wider windows, thus improving image quality. Conversely, the OC in the parenchymal phase scans must have a high gradient in order to better differentiate between the vascular structures and the surrounding hepatic parenchyma. CONCLUSIONS: Image features considered to be of interest by the operator cannot be simplified by the mean of pre-set OC. Due to their strong individual variability automatic 3D algorithms cannot be universally applied: they should be adapted to both image and patient characteristics. PMID- 15883523 TI - Intra and interobserver variability of renal allograft ultrasound volume and resistive index measurements. AB - PURPOSE: Aim of the present study was to evaluate the repeatability and reproducibility of the Doppler Resistive Index (R.I.) and the Ultrasound renal volume measurement in renal transplants. MATERIALS AND METHODS: Twenty-six consecutive patients (18 men, 8 women) mean age of 42,8+/-12,4 years (M+/-SD) (range 22-65 years) were studied twice by each of two trained sonographers using a color Doppler ultrasound scanner. Twelve of them had a normal allograft function (defined as stable serum creatinine levels = or <123,76 micromol/L), whilst the remaining 14 had decreased allograft function (serum creatinine 132.6 265.2 micromol/L). Results were given as mean of 6 measurements performed at upper, middle and lower pole of the kidney. Intra- and interobserver variability was assessed by the repeatability coefficient and coefficient of variation (CV). RESULTS: Regarding Resistive Index measurement, repeatability coefficient was between 0.04 and 0.06 and the coefficient of variation was <5%. The analysis of the Student's t test did not show any significant difference between the measurements (t=0.15; p=0.87 n.s.). A good reproducibility was also detected in US measurements of renal length and volume. CONCLUSIONS: These results suggest that Color Doppler Resistive Index measurements of renal allograft and Ultrasound renal volume measurements are repeatable and reproducible. PMID- 15883524 TI - MRI versus scintigraphy with 99mTc-HMPAO-labelled granulocytes in the diagnosis of bone infection. AB - PURPOSE: To assess the reliability of MRI and scintigraphy with 99mTc-HMPAO labelled granulocytes in the diagnosis of bone infection. MATERIALS AND METHODS: Between November 2001 and November 2003, 25 patients (16 men and 9 women; age range 22-72 years; mean age 48 years) with suspected bone infection were evaluated. The lower limbs were more frequently involved (20/25 cases). MRI (T1 and T2-w sequences, both with and without fat suppression; T1-w fat-suppressed sequence after Gadolinium administration) and scintigraphy with 99mTc-HMPAO labelled granulocytes were performed in all patients; the study was performed 30 min, 150 min and 24 h after the injection of 99mTc labelled autologous granulocytes. The maximum interval between MRI and scintigraphy was 15 days. The diagnosis was confirmed surgically (18/25 cases) or by clinical follow-up (7/25 cases). RESULTS: In 10/25 cases the clinical suspicion of bone infection was confirmed by MRI, scintigraphy and surgery. In 12/25 cases the clinical suspicion of bone infection was not confirmed either by imaging, surgery (5/12 cases) or follow-up (7/12 cases). In 3/25 cases the results were discordant: in 1/3 cases with chronic osteomyelitis confirmed by MRI and surgery, a false-negative diagnosis was made by scintigraphy. In 2/3 cases with negative MRI diagnoses confirmed by surgery, a false-positive diagnosis was made by scintigraphy owing to the infection of adjacent soft tissues. CONCLUSIONS: MRI is more reliable for the detection and evaluation of the local spread of bone infection. Conversely, when metallic devices causing artefacts on MR images are present, scintigraphy is the preferred alternative modality. PMID- 15883525 TI - Contribution of imaging to the evaluation of pyomyositis. AB - PURPOSE: To analyse the fundamental features of each imaging modality with the aim of defining their contribution to the diagnosis of pyomyositis. MATERIALS AND METHODS: We retrospectively reviewed 10 observations collected between 1997 and 2004, 5 of paravertebral muscles and 5 of the muscles of the limbs. Imaging was carried out with US (8/10 observations) and/or CT (9/10 observations), and/or MRI (6/10 observations). The pathogen was isolated in 6/10 observations. RESULTS: The imaging criteria for the detection and characterization of pyomyositis are based on three pathological phases: an invasive phase characterized by inflammatory oedema, a suppurative phase characterised by abscess development, and a late phase featuring the distant spread of the infection. Based on these interpretation criteria, pyomyositis was correctly diagnosed with US in 5/8 observations, with CT in 9/9 observations and with MR in 5/6 observations. Four false-negative cases were found. CONCLUSIONS: Since the clinical symptoms of pyomyositis are often insidious and delayed, an integrated imaging approach may be very useful for diagnosing this condition. PMID- 15883526 TI - Contribution to the radiological study of the eosinophilic granuloma of the mandible (unifocal granuloma due to Langerhans' cell histiocytosis). AB - PURPOSE: The radiological diagnosis of osteolytic lesions of the mandible still constitutes a challenge in some pathological conditions in which the clinical data and the case history are relatively uniform and the radiological picture is lacking in any characteristics. MATERIALS AND METHODS: We reviewed the conventional radiograms of six cases of Langerhans' cell histiocytosis (LCH) of the mandible examined over the last ten years. The X-ray examinations were performed in the lateral-oblique projection to allow a view of the horizontal portion of the mandible almost completely free of overlapping images of other bone structures. RESULTS: We identified a series of radiological patterns for these reticulo-endotheliopathies capable of causing granuloma formed by polynuclear eosinophils, plasma cells, lymphocytes, and large mononuclear macrophages with granulopexic and phagocytic activity that proliferate in the bone tissue and can be identified as the Langerhans' cells of skin, mucosa, periodontal cavities and bone marrow. The granulomatous tissue penetrates the affected organ elements and, in its spread, it compresses, atrophies, and destroys the damaged tissue, replacing it. The alterations produced by Langerhans' cell histiocytic granuloma are most common in the skeletal system affecting, in order of frequency, the skull, the long bones of the limbs, the foot, the ribs and the spine. The oral mucosa is rarely involved. CONCLUSIONS: The radiological investigation of unifocal Langerhans' cell histiocytic granuloma of the mandible is essential in the study of perimandibular swelling, although diagnosis is based on biopsy alone. In addition to digital or conventional radiography, other useful examinations are bone scintigraphy, Colour Doppler US, MR and CT, which enables a correct localisation necessary for planning the biopsy and treatment. PMID- 15883527 TI - Sacral-neuromodulation CT-guided. AB - PURPOSE: Sacral neuromodulation is a new treatment for refractory voiding disorders such as urge incontinence, urinary retention, frequency-urgency syndromes and faecal incontinence. The current approach to sacral nerve stimulation consists of a two-stage procedure. The first is a PNE test (Percutaneous Nerve Evaluation) by a provisional electrically stimulated spinal needle, placed percutaneously in the S3 foramina for four to ten days. If successful, the second stage, permanent implantation, is carried out. The PNE test is performed under fluoroscopic control using the palpable bony sacral foramina as referral points. This technique can show some limitations, such as operator Rx exposure, poor visualization of sacral foramina because of bowel gas artefacts or sacral malformation. In order to reduce these inconveniences and to improve efficiency of the test we tried an alternative technique. The purpose of our study was to test the use of CT as an alternative technique in order to evaluate its advantages and possible routine use. MATERIALS AND METHODS: We tested 30 patients with the PNE test under CT guidance (16 males and 14 females) suffering from serious pelvic disorders and not responding to the normal therapeutic regime. Twenty-seven patients showed relative anatomical integrity of the pelvis and the sacrum, the remaining 3 patients presented morphological anomalies of the sacral foramina. With the patient in the prone position the sacral foramina were identified with CT volumetric scanning using a spiral CT scanner equipped with a second console for the three-dimensional reconstructions. Having identified the location of the S3 foramina, a sterile field was prepared and the spinal needle introduced checking correct positioning with a CT control scan. An electrode was then inserted after having checked correct muscular contractile response and the precise position with a further CT scan. RESULTS: Thirty patients were subjected to PNE under CT guidance for a total of 38 centerings. Eight patients underwent the PNE procedure on both the S3 foramina. The sacral foramen was centred at the first attempt in 36 out of 38 cases. Two cases required several attempts to centre correctly the foramen. In 4 patients out of 30 a second electrode was implanted. In one patient who had a nonconsolidated sacral fracture, CT guidance enabled insertion of the electrode inside the only practicable foramen, a manoeuvre that would have been impossible with fluoroscopical guidance. Only once was the electrode placed in a wrong location but promptly repositioned after a CT control. During the whole trial period we had a positive response to the PNE test in 18 out of 30 patients (60%), a partial response in 4 out of 30 patients and a negative response in the remaining 8 patients. None of the patients who underwent the PNE test had infectious complications and the procedure was well tolerated by all. The procedure lasted about 45 minutes. PMID- 15883528 TI - Lumbar sympathectomy under CT guidance: therapeutic option in critical limb ischaemia. AB - PURPOSE: Lumbar sympathectomy is a complementary therapeutic option for patients with severe peripheral vascular occlusive disease presenting rest pain or gangrene and not eligible for surgical revascularisation. Traditional surgical sympathectomy was widely used in the past. However, due to its invasive character, it has increasingly been replaced by percutaneous techniques and, in some recent cases, by laparoscopic procedures. Percutaneous lumbar sympathectomy is a safe, cost-effective and widely available treatment option. We report our experience on 19 patients subjected to percutaneous sympathectomy under CT guidance. MATERIALS AND METHODS: Between 1998 and 2000, 19 patients underwent percutaneous sympathectomy under CT guidance. All patients had severe vascular disease of the lower extremities (Fontaine stage IV), with rest pain and gangrene. They were not eligible for surgical revascularisation. Phenol was injected at the level of L2 and L4 using two 22 G needles (15 cm long). Signs of interrupted sympathetic activity usually occur 2'-15' after the procedure with warmth and flushing and dryness of the lower extremities. RESULTS: Percutaneous sympathectomy under CT guidance is a simple, safe and well-tolerated procedure with a low rate of complications. Of the 19 patients, 9 (47.3%) showed clinical improvement, whereas 5 experienced a worsening of ischaemia in the months immediately following the procedure. DISCUSSION: Results suggest that percutaneous lumbar sympathectomy causes a sympathetic blockade in patients with advanced vascular disease of the limbs. CT guidance ensures a high level of precision in drug dosing, thus lowering the risk of complications. Although the results are demoralizing. the impossibility of achieving surgical revascularisation in advanced peripheral arteriosclerosis enhances the role of CT guided percutaneous sympathectomy in relieving rest pain and healing ulcers in order to postpone the amputation. PMID- 15883541 TI - National Institutes of Health ushers in new age of open-access publishing. PMID- 15883542 TI - Appreciative leadership. PMID- 15883543 TI - Advocacy for coverage of nutrition services. PMID- 15883544 TI - Methodological issues in assessing dietary supplement use in children. PMID- 15883545 TI - Predictors of improvement in hemoglobin concentration among toddlers enrolled in the Massachusetts WIC Program. AB - OBJECTIVE: Nutrition supplementation programs are generally targeted to those members of the population who are thought to be at risk of an undesirable outcome, but not all who participate in such programs respond to them. We sought to identify determinants of improvement in hemoglobin concentration among young children in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). DESIGN: We conducted an observational study using data from 9,930 children who were enrolled in the Massachusetts WIC program and had data available on their hemoglobin values at both 1 and 2 years of age. Predictors of change in hemoglobin concentration between these ages were studied using multivariate statistical modeling. RESULTS: Overall, hemoglobin concentrations increased from age 1 to 2 years in those who had been breastfed 25 or more weeks (P < .0001) and were female (P < .01), and decreased with increasing weight at 1 year of age (P < .001). The determinants of change in hemoglobin concentration differed from the determinants of hemoglobin concentration at age 1 year. CONCLUSIONS: The analytical approach used here could be extended to identify subgroups of WIC participants likely to improve in other outcomes. If current efforts to increase the duration of breastfeeding among WIC participants are successful, the importance of WIC in improving hemoglobin concentration among young children also will increase. PMID- 15883546 TI - Promoting breastfeeding to mothers in the Special Supplemental Nutrition Program for Women, Infants, and Children. PMID- 15883547 TI - Changes in growth and seizure reduction in children on the ketogenic diet as a treatment for intractable epilepsy. AB - OBJECTIVE: To assess growth and seizure reduction in epileptic children using the ketogenic diet as a treatment for intractable epilepsy. DESIGN: A retrospective chart review was designed to evaluate urinary ketone levels, height and weight z scores and percentiles, and seizure reduction. Data were collected at baseline and at 6 and 12 months. SUBJECTS/SETTINGS: Fifty-seven subjects, ages 1 to 26 years old, started the ketogenic diet at Rush University Medical Center between August 1995 and December 2001. Thirty-nine subjects stayed on the diet for 6 months, and five more were lost by the 12th month of follow-up. Statistical analysis Mann-Whitney U tests assessed differences between male and female subjects and between subjects with high ketosis and moderate ketosis. Friedman tests followed by Wilcoxon sign rank tests assessed the significance of changes in growth at baseline and at 6 and 12 months. Associations between seizure reduction and growth and urinary ketone levels were determined using chi 2 tests. A binary logistic regression model identified potential predictors of growth and seizure reduction. RESULTS: Height-for-age z scores significantly decreased ( P < or =.0005) from -0.30+/-1.19 to -0.99+/-1.13 among subjects on the diet for 12 months. Subjects with high ketosis (80 to 160 mg/dL) experienced a significant decrease ( P < or =.0005) in height-for-age z scores from -0.45+/-1.28 to -1.1+/ 1.23, whereas subjects with moderate ketosis did not. Observed percent seizure reduction was similar to those of other published studies. CONCLUSIONS: Subjects on the ketogenic diet showed a delay in growth. More research is needed to evaluate the relationship between ketosis and growth. PMID- 15883548 TI - Children and the ketogenic diet. PMID- 15883549 TI - Relative validity and reliability of a food frequency questionnaire for a triethnic population of 1-year-old to 3-year-old children from low-income families. AB - OBJECTIVE: To develop and validate a food frequency questionnaire (FFQ) for low income, 1- to 3-year-old Hispanic, African-American, and white children. DESIGN: A convenience sample of low-income mothers provided dietary data for their child via FFQs and 3-day diet records. SUBJECTS/SETTING: Participants (N=77) were recruited from Special Supplemental Program for Women, Infants, and Children clinics and public health clinics. All subjects were Hispanic, African-American, or white mothers of children 1 to 3 years old, 18 years of age or older, income less than 200% of the federal poverty level, and were literate in English. Statistical analyses performed Wilcoxon signed-ranks tests were used for comparisons of food group servings on time 1 and time 2 FFQs and to examine differences in food group servings between the FFQ and diet records. Spearman correlations were calculated to assess test-retest reliability and the relative validity. Contingency tables were used to determine the degree of association between the FFQ and diet records. RESULTS: Reliability correlations were significant for all nine food categories ( =0.69), ranging from 0.53 (soups) to 0.84 (nonstarchy vegetables). Validity correlations were significant for all food groups ( =0.41) except starchy vegetables. Thirty-six percent of children were classified into the same quartile of food group intake and 78% into the same or within one quartile. CONCLUSIONS: The FFQ yielded excellent reliability and acceptable validity and can be used to assess food choices in a triethnic sample of low-income 1- to 3-year-old children. PMID- 15883550 TI - The relationship of dietary and lifestyle factors to bone mineral indexes in children. AB - OBJECTIVE: To identify factors related to children's bone mineral indexes at age 8 years, and to assess bone mineral indexes in the same children at ages 6 and 8 years. DESIGN: Bone mineral content (BMC [g]) and bone mineral density (BMD; calculated as g/cm 2 ) were measured by dual-energy x-ray absorptiometry (DEXA) in children and their mothers when the children were 8 years of age. A subset of children had an earlier DEXA assessment at age 6 years. Children's dietary intake, height, weight, and level of sedentary activity were assessed as part of a longitudinal study from ages 2 months to 8 years. SUBJECTS/SETTING: Fifty-two healthy white children (25 male, 27 female) and their mothers. Main outcome measures Children's total BMC and BMD at age 8 years. STATISTICAL ANALYSES PERFORMED: Correlations and stepwise multiple regression analyses. RESULTS: Factors positively related to children's BMC at age 8 years included longitudinal intakes (ages 2 to 8 years) of protein, phosphorus, vitamin K, magnesium, zinc, energy, and iron; height; weight; and age ( P < or = .05). Factors positively related to children's BMD at age 8 years included longitudinal intakes of protein and magnesium ( P < or = .05). Female sex was negatively associated with BMC and BMD at age 8 years ( P < or = .05). Children's bone mineral indexes at ages 6 and 8 years were strongly correlated ( r =0.86, P < .0001 for BMC; r =0.92, P <.0001 for BMD). CONCLUSIONS: Because many nutrients are related to bone health, children should consume a varied and nutrient-dense diet. PMID- 15883552 TI - Breakfast habits, nutritional status, body weight, and academic performance in children and adolescents. AB - Breakfast has been labeled the most important meal of the day, but are there data to support this claim? We summarized the results of 47 studies examining the association of breakfast consumption with nutritional adequacy (nine studies), body weight (16 studies), and academic performance (22 studies) in children and adolescents. Breakfast skipping is highly prevalent in the United States and Europe (10% to 30%), depending on age group, population, and definition. Although the quality of breakfast was variable within and between studies, children who reported eating breakfast on a consistent basis tended to have superior nutritional profiles than their breakfast-skipping peers. Breakfast eaters generally consumed more daily calories yet were less likely to be overweight, although not all studies associated breakfast skipping with overweight. Evidence suggests that breakfast consumption may improve cognitive function related to memory, test grades, and school attendance. Breakfast as part of a healthful diet and lifestyle can positively impact children's health and well-being. Parents should be encouraged to provide breakfast for their children or explore the availability of a school breakfast program. We advocate consumption of a healthful breakfast on a daily basis consisting of a variety of foods, especially high-fiber and nutrient-rich whole grains, fruits, and dairy products. PMID- 15883554 TI - Longitudinal patterns of vitamin and mineral supplement use in young white children. AB - OBJECTIVES: The objective of this article is to report longitudinal patterns of nutrient supplementation in children, to quantify nutrient intakes from supplements and diet, and to examine relationships between supplement use and sociodemographic factors. DESIGN AND SUBJECTS: Subjects were followed prospectively from birth to 24 months with periodic questionnaires on vitamin/mineral supplement use. Food diaries were completed to report food and beverage intake. Estimates of daily intakes of vitamins and minerals were calculated. Fisher's Exact tests and t tests were used to assess the association of sociodemographic variables with supplement use. RESULTS: A substantial proportion of young children used supplements. The prevalence of supplement use increased with age. By the end of 24 months, 31.7% used some supplement. Young supplement users consume supplements regularly, ranging from 40% to 60% of days reported. The majority of young children in this cohort could obtain adequate vitamins and minerals from diet alone for the first 24 months of life. Intakes of some nutrients from diet alone, such as vitamin E and folate, were not sufficient for a large proportion of young children. However, intake above the dietary reference intake was observed for a few nutrients, in particular for vitamin A. CONCLUSIONS: Use of nutrient supplements is a common behavior during the first 2 years of life. This study shows that most young children obtain adequate nutrients from diet alone. Health professionals should provide recommendations for nutrient supplementation of generally healthy children based on an assessment of their dietary practices. PMID- 15883556 TI - Comparison of predictive equations for resting metabolic rate in healthy nonobese and obese adults: a systematic review. AB - BACKGROUND: An assessment of energy needs is a necessary component in the development and evaluation of a nutrition care plan. The metabolic rate can be measured or estimated by equations, but estimation is by far the more common method. However, predictive equations might generate errors large enough to impact outcome. Therefore, a systematic review of the literature was undertaken to document the accuracy of predictive equations preliminary to deciding on the imperative to measure metabolic rate. METHODS: As part of a larger project to determine the role of indirect calorimetry in clinical practice, an evidence team identified published articles that examined the validity of various predictive equations for resting metabolic rate (RMR) in nonobese and obese people and also in individuals of various ethnic and age groups. Articles were accepted based on defined criteria and abstracted using evidence analysis tools developed by the American Dietetic Association. Because these equations are applied by dietetics practitioners to individuals, a key inclusion criterion was research reports of individual data. The evidence was systematically evaluated, and a conclusion statement and grade were developed. RESULTS: Four prediction equations were identified as the most commonly used in clinical practice (Harris-Benedict, Mifflin-St Jeor, Owen, and World Health Organization/Food and Agriculture Organization/United Nations University [WHO/FAO/UNU]). Of these equations, the Mifflin-St Jeor equation was the most reliable, predicting RMR within 10% of measured in more nonobese and obese individuals than any other equation, and it also had the narrowest error range. No validation work concentrating on individual errors was found for the WHO/FAO/UNU equation. Older adults and US residing ethnic minorities were underrepresented both in the development of predictive equations and in validation studies. CONCLUSIONS: The Mifflin-St Jeor equation is more likely than the other equations tested to estimate RMR to within 10% of that measured, but noteworthy errors and limitations exist when it is applied to individuals and possibly when it is generalized to certain age and ethnic groups. RMR estimation errors would be eliminated by valid measurement of RMR with indirect calorimetry, using an evidence-based protocol to minimize measurement error. The Expert Panel advises clinical judgment regarding when to accept estimated RMR using predictive equations in any given individual. Indirect calorimetry may be an important tool when, in the judgment of the clinician, the predictive methods fail an individual in a clinically relevant way. For members of groups that are greatly underrepresented by existing validation studies of predictive equations, a high level of suspicion regarding the accuracy of the equations is warranted. PMID- 15883557 TI - Dietary intakes at child-care centers in central Texas fail to meet Food Guide Pyramid recommendations. AB - The purpose of this study was to compare the dietary intakes of children who attend child-care centers with the recommendations of the Food Guide Pyramid for Young Children. Three-day dietary records were obtained for 50 children ages 3 to 5 years old who attended nine child-care centers in central Texas. Dietary intakes during child care were measured by a researcher, and dietary intakes at home were recorded by a parent. During child care, the 3-year-old children consumed sufficient fruits and meat/alternates, but not sufficient grains, vegetables, or dairy to meet two thirds of Food Guide Pyramid for Young Children recommendations. The 4- and 5-year-old children only consumed sufficient dairy. Ninety-one percent (20 of 22) of the 3-year-old children met two thirds of their estimated energy requirements during child care, compared with 5% (1 of 20) of 4 year-old children and 25% (2 of 8) of 5-year-old children. Intakes at home did not compensate for lack of grain and vegetable consumption during child care. PMID- 15883558 TI - Determination of body composition in children with cerebral palsy: bioelectrical impedance analysis and anthropometry vs dual-energy x-ray absorptiometry. AB - The object of this study was to determine whether bioelectrical impedance analysis (BIA) and anthropometry can be used to determine body composition for clinical and research purposes in children with cerebral palsy. Eight individuals with cerebral palsy (two female, mean age=10 years, mean gross motor function classification=4.6 [severe motor impairment]) recruited from an outpatient tertiary care setting underwent measurement of fat mass, fat-free mass, and percentage body fat using BIA, anthropometry (two and four skinfold equations), and dual-energy x-ray absorptiometry. Correlation coefficients were calculated for fat mass, fat-free mass, and percent body fat for these measures as determined by BIA and anthropometry when compared with dual-energy x-ray absorptiometry. Correlation coefficients were excellent for determination of fat free mass for all methods (ie, all were above 0.9). Correlations were moderate for determination of fat mass and percent body fat (range=0.4 to 0.8). PMID- 15883559 TI - Few differences found in the typical eating and physical activity habits of lower level and upper-level university students. AB - Typical eating and exercise habits of a group of lower-level (freshmen/sophomores, n=144) and upper-level (juniors/seniors, n=114) students, 19 to 25 years of age, attending a midwestern university were compared. Both groups reported similar snacking practices, nonalcoholic beverage consumption, places of meal consumption, factors influencing food choices, duration and frequency of various types of physical activity, and places of physical performance, with few exceptions. Significant differences between groups were observed for typical afternoon snack consumption, frequency of eating in university cafeterias, typical time spent walking, typical frequency of performing other aerobic activities, and factors that most influenced their physical activity habits. Few differences existed in typical eating and physical activity habits of lower- and upper-level students. PMID- 15883560 TI - Validation of a brief diet survey instrument among medical students. AB - Our objective was to assess the reproducibility and accuracy of fat and of fruit and vegetable items on a 43-item food frequency questionnaire (FFQ) previously self-administered by students at 16 US medical schools. Five in-person, 24-hour recalls were administered between two FFQ administrations to 88 medical students. Reported fat intake decreased from the first (34.7%) to the second (33.1%) FFQ administration ( P <.001); the reproducibility correlation was r =0.63. Fat intake from recalls (28.4%) was lower than that from the FFQ (33.8%, P <.001). The Pearson correlation was r =0.36. Fruit and vegetable servings per day were 3.9 and 3.7 from the first and second FFQ, respectively ( P =.5); the reproducibility correlation was r =0.77. Fruit and vegetable servings were marginally higher from recalls (4.3) than from the FFQ (3.8, P =.06). The Pearson correlation for fruit and vegetable servings was r =0.50. This brief FFQ provides acceptably reproducible and valid estimates of fruit and vegetable servings per day among most groups of medical students, but overestimates fat as a percentage of energy intake. PMID- 15883561 TI - Calcium fortification systems differ in bioavailability. AB - The objective of this study was to compare the bioavailability of calcium from two fortification systems used in orange juice. The design was randomized crossover, within-subject. The subjects were 25 healthy premenopausal women in an academic health sciences center. Two commercially marketed calcium-fortified orange juices, ingested in an amount providing 500 mg calcium, were taken at breakfast after an overnight fast. The two fortification systems tested were calcium citrate malate and a combination of tricalcium phosphate and calcium lactate (tricalcium phosphate/calcium lactate). The main outcome measure was the area under the curve (AUC) for the increase in serum calcium from 0 to 9 hours after ingesting the test calcium source. Statistical analyses performed were repeated measures analysis of variance, testing source, and sequence. AUC 9 was 48% greater for calcium citrate malate than for tricalcium phosphate/calcium lactate ( P < .001); absorbed calcium calculated from AUC 9 values (mean+/ standard error of the mean) was 148+/-9.0 mg and 100+/-8.9 mg for calcium citrate malate and tricalcium phosphate/calcium lactate, respectively. The results indicate that equivalent calcium contents on a nutritional label do not guarantee equivalent nutritional value. Nutritionists and dietetics professionals should encourage manufacturers of fortified products to provide information on bioavailability. PMID- 15883562 TI - Position of the American Dietetic Association: Promoting and supporting breastfeeding. AB - It is the position of the American Dietetic Association (ADA) that exclusive breastfeeding provides optimal nutrition and health protection for the first 6 months of life, and breastfeeding with complementary foods for at least 12 months is the ideal feeding pattern for infants. Breastfeeding is also a public health strategy for improving infant and child health survival, improving maternal morbidity, controlling health care costs, and conserving natural resources. ADA emphasizes the essential role of dietetics professionals in promoting and supporting breastfeeding by providing up-to-date, practical information to pregnant and postpartum women, involving family and friends in breastfeeding education and counseling, removing institutional barriers to breastfeeding, collaborating with community organizations and others who promote and support breastfeeding, and advocating for policies that position breastfeeding as the norm for infant feeding. ADA also emphasizes its own role by providing up-to-date information to the public, encouraging empirical research, providing continuing education opportunities, providing cultural sensitivity and cultural competence training to dietetics professionals, and encouraging universities to review and update undergraduate and graduate training programs. PMID- 15883563 TI - American Dietetic Association: Standards of practice and standards of professional performance for registered dietitians (generalist, specialty, and advanced) in diabetes care. PMID- 15883564 TI - Medical nutrition therapy: the core of ADA's advocacy efforts (part 1). PMID- 15883565 TI - Medication effects on metabolic rate: a systematic review (part 1). PMID- 15883570 TI - Symptomatic Arnold-Chiari malformation and cranial nerve dysfunction: a case study of applied kinesiology cranial evaluation and treatment. AB - OBJECTIVE: To present an overview of possible effects of Arnold-Chiari malformation (ACM) and to offer chiropractic approaches and theories for treatment of a patient with severe visual dysfunction complicated by ACM. CLINICAL FEATURES: A young woman had complex optic nerve neuritis exacerbated by an ACM type I of the brain. INTERVENTION AND OUTCOME: Applied kinesiology chiropractic treatment was used for treatment of loss of vision and nystagmus. After treatment, the patient's ability to see, read, and perform smooth eye tracking showed improvement. CONCLUSION: Further studies into applied kinesiology and cranial treatments for visual dysfunctions associated with ACM may be helpful to evaluate whether this single case study can be representative of a group of patients who might benefit from this care. PMID- 15883571 TI - Pressures generated during spinal manipulation and their association with hand anatomy. AB - BACKGROUND CONTEXT: The role of the variation in the application manipulation itself is largely unknown. A greater understanding of its input parameters is necessary to better understand spinal manipulation outcomes. PURPOSE: The objective of this study is to determine if pressures generated during manipulation are altered by hand configuration. DESIGN/SETTING: Paired comparison of 2 different variable groups. METHODS: Sixteen chiropractors provided 2 manipulations to a rigid surface using 2 hand configurations used commonly in clinical practice: arched and flat. Interposed between the hand and the rigid surface was a pressure sensor array and radiographic cassette. For each manipulation, pressures were recorded and a radiographic image was captured. Two radiologists then located the osseous features of the hand with respect to the sensor array. RESULTS: In 15 of 16 cases, arched configurations produced peak pressures that corresponded to the radiographic location of the pisiform bone. In flat configurations, peak pressure migrated about the location of the hamate bone. Radiologists' agreement for bone position was high (kappa = 0.96). Measures of peak pressure, total pressure, and pressure distribution were statistically different between hand configurations. CONCLUSIONS: The results of this study suggest that hand configuration influences the magnitude, location, and distribution of pressure generated by the hand during manipulation. This knowledge may have importance in understanding the relation among application parameters of manipulation, therapeutic benefit, and patient safety. PMID- 15883572 TI - Evidence-based care, certainty, and the doctor's duty of care. PMID- 15883574 TI - The Bournemouth Questionnaire: can it be used to monitor and predict treatment outcome in chiropractic patients with persistent low back pain? AB - OBJECTIVE: To investigate the Bournemouth Questionnaire (BQ) as a baseline, monitoring of progress, and prognostic instrument in chiropractic patients with persistent low back pain (LBP). STUDY DESIGN: Predictive and concurrent validation study. STUDY PARTICIPANTS AND SETTING: One hundred fifteen Norwegian chiropractors collected prospective data on 875 patients with persistent LBP, defined as LBP for at least 2 weeks at baseline and a minimum of 30 days totaling within the preceding year. METHODS: Data collection took place at first consultation, fourth visit, and 3 months using the BQ, the revised Oswestry questionnaire, and a 10-point pain box scale. Follow-up at 12 months included the BQ, Oswestry questionnaire, and additional questions on the number of days with LBP and the number of days off work in the past year. DATA ANALYSIS: Frequency of reporting of each 7 items in the BQ at baseline was identified as median value with 10th and 90th percentiles. Concurrent analyses of the 2 questionnaires were made at the 4 points in time with calculation of mean differences with limits of agreement together with Bland-Altman plots. Logistic regression was used to identify and compare the predictive values of the questionnaires and to test the relevance of each individual item in the BQ. RESULTS: The median baseline values of the 7 items in the BQ ranged from 2 to 5. The 2 questionnaires did not agree on patients' status, and mean differences between the Oswestry questionnaire and the BQ were largest when patients reported higher scores. The predictive values for the 2 questionnaires were low, with no significant difference between the 2. The predictive value of the BQ could be improved by removing most of the 7 items. Certain items can predict specific outcomes. CONCLUSIONS: The BQ is not a useful instrument to identify baseline status, monitor progress, or predict the 1-year progress in chiropractic patients having persistent LBP. However, certain individual items are useful to predict specific outcomes. PMID- 15883575 TI - Chronic disease self-management program for low back pain in the elderly. AB - OBJECTIVE: To evaluate the effectiveness of Stanford's Chronic Disease Self Management Program (CDSMP) for chronic low back pain (LBP) in older Americans. DESIGN: Randomized controlled trial. SETTING: Community-based program offered at 12 locations. SUBJECTS: Community-dwelling seniors (n = 109) aged 60 and older with chronic LBP of mechanical origin. METHODS: Patients were randomly allocated to the CDSMP or to a 6-month, wait-list control group. The program included one 2.5-hour session per week for 6 weeks. Outcomes evaluated at 6 months included 100-point modified Von Korff pain and disability scales; days with pain and disability; SF-36 general health, energy-fatigue, and emotional well-being scales; 2 scales from the Arthritis Self-Efficacy Scale, self-care attitudes/behaviors, and health services utilization. RESULTS: For pain at 6 months, the primary outcome, the adjusted mean difference between the program and control, was -1.0 (P = .835). There was a sizable advantage for the program in disability averaged over the course of the entire 6-month study (-9.2, P = .027), but not at the 6-month follow-up (-5.8, P = .278). There was an interaction between intervention and baseline disability days favoring the program for higher baseline values (P = .007). The CDSMP affected emotional well-being (7.6, P = .037) and energy-fatigue (5.1, P = .274). There were no differences for self efficacy, pain days, and general health. CONCLUSION: There was no advantage for the CDSMP over a wait-list control for improving pain, general health, self efficacy, and self-care attitudes in older Americans with chronic LBP. A benefit was suggested for emotional well-being, fatigue, functional disability, and days with disability. PMID- 15883576 TI - Measurement of lumbar spine loads and motions during rotational mobilization. AB - OBJECTIVES: To measure the loads acting at the lumbar spine and the resulting motions during rotational mobilization. METHODS: Twenty healthy subjects were subjected to right rotational mobilization of different grades. The magnitude and frequency of mobilization loads were decided by an experienced manipulative therapist to be appropriate for each mobilization grade. Subjects were positioned on a specially constructed plinth capable of measuring forces and moments about 3 axes. The 3-dimensional movements of the lumbar spine were captured by an electromagnetic tracking device. RESULTS: In the starting positions, the lumbar spines were found to be flexed, axially rotated to the right, and laterally bent to the left. As the mobilization grade increased, the spine was axially rotated further into the range. Rotational mobilization was found to induce oscillatory moments and movements of the spine in all 3 anatomical planes. The twisting moment and movement were generally accompanied by lateral bending moment and movement in the opposite direction. The mean amplitudes of the moment and movement oscillations were small and found to be largest for grade III mobilization. CONCLUSION: The mechanical effects of rotational mobilization are not restricted to axial rotation of the spine as the name may have suggested. Rotational mobilization may be able to restore lost movements of the lumbar spine in any of the 3 planes. The method developed in this study showed good reliability and may be considered to assess treatment outcome and changes in spinal stiffness after therapy. PMID- 15883577 TI - Supplemental care with medication-assisted manipulation versus spinal manipulation therapy alone for patients with chronic low back pain. AB - OBJECTIVES: To measure changes in pain and disability for chronic low-back pain patients receiving treatment with medication-assisted manipulation (MAM) and to compare these to changes in a group only receiving spinal manipulation. STUDY DESIGN: Prospective cohort study of 68 chronic low-back pain patients. METHODS: Outcomes were measured using the 1998 Version 2.0 American Association of Orthopaedic Surgeons/Council of Musculoskeletal Specialty Societies/Council of Spine Societies Outcomes Data Collection Instruments. The primary outcome variable was change in pain and disability. All patients received an initial 4- to 6-week trial of spinal manipulation therapy (SMT), after which 42 patients received supplemental intervention with MAM and the remaining 26 patients continued with SMT. RESULTS: Low back pain and disability measures favored the MAM group over the SMT-only group at 3 months (adjusted mean difference of 4.4 points on a 100-point scale, 95% confidence interval [CI] -2.2 to 11.0). This difference attenuated at 1 year (adjusted mean difference of 0.3 points, 95% CI 8.6 to 9.2). The relative odds of experiencing a 10-point improvement in pain and disability favored the MAM group at 3 months (odds ratio 4.1, 95% CI 1.3-13.6) and at 1 year (odds ratio 1.9, 95% CI 0.6-6.5). CONCLUSION: Medication-assisted manipulation appears to offer some patients increased improvement in low back pain and disability. Further investigation of these apparent benefits in a randomized clinical trial is warranted. PMID- 15883578 TI - Geographical distribution of judo therapists and orthopedists in Japan. AB - BACKGROUND: Judo therapy is a unique and long-established Japanese comedical profession, specialized in manually treating fractures and sprains in practitioners' offices. Having the overlapped scopes of services between judo therapists and orthopedists, disputes between the 2 have gradually emerged recently. OBJECTIVE: To examine the relationship between judo therapists and orthopedists by comparing their geographic distributions at the whole country level and at prefecture level. STUDY DESIGN: Cross-sectional study. METHODS: The numbers of judo therapist facilities and orthopedists per 100,000 population in each municipality were obtained through web yellow pages and from the Japanese census data. Lorenz curves and Gini indices were calculated to show the distributions and to compare the numbers of judo therapist facilities and orthopedists per 100,000 population. RESULTS: The median number of judo therapist facilities per 100,000 population in all municipalities was 12.0 (range 0-135) and that for orthopedists was 4.3 (0-186). The Gini indices of the nationwide judo therapist facilities and those for orthopedists were 0.29 and 0.36, respectively. CONCLUSION: These data show the relatively homogeneous distribution of judo therapist facilities compared with orthopedists. In smaller municipalities, judo therapist facilities may fill in the absence of orthopedists. As for lager municipalities, further study is needed to examine whether the relationships between them are competitive or complementary. PMID- 15883579 TI - Treatment of chronic nonresponsive patients with a nonforce technique. AB - OBJECTIVE: To investigate how chronic pain patients respond to treatment with Bio Energetic Synchronization Technique (BEST). METHODS: Twenty-four adult patients with chronic pain-related conditions that failed to respond to previous chiropractic care were recruited. Subjects were given baseline assessments including pain Visual Analog Scale, Profile of Mood States, and the Global Well being Scale. The 5-week treatment program consisted of an initial 3-day session with BEST therapy, followed by a single treatment session for the following 4 weeks. Patients were reevaluated at the end of the 3-day session and at weekly intervals throughout the course of care. At the end of week 5, patients were asked to assess their degree of satisfaction with the treatment. RESULTS: Patients had 3 main categories of pain: headache (n = 8, mean duration 15 years), neck pain (n = 18, mean duration 11 years), and low back pain (n = 17, mean duration 10 years). Global Well-Being Scale scores significantly improved at the end of the 3-day session (P > .05) but not subsequently. The Profile of Mood States reflected favorable changes in all areas. Significant improvement in vigor (P > .003) and fatigue (P > .006) existed at the end of 5 weeks (P < .01). The reduction of pain was significant at both the end of the 3-day session and at follow-up (P = .0003). A statistically significant decrease in depression (P = .004) was noted after 3 days, and a substantial although not significant (P = .06) decrease in depression existed at the end of 1 month. Eighty-two percent reported satisfaction with BEST (47% reported being "extremely satisfied" and 35% "satisfied"). CONCLUSION: In this group of chronic pain patients, improvement in patient outcome measures was seen after 5 weeks of therapy. These patients also responded with a high degree of satisfaction with care. PMID- 15883580 TI - Distraction manipulation of the lumbar spine: a review of the literature. AB - OBJECTIVE: The purpose of this study is to review the literature concerning distraction manipulation of the lumbar spine, particularly regarding physiological effects, clinical efficacy, and safety. DATA SOURCES: A search of the English language literature was conducted using the MEDLINE, Embase, CINAHL, Chiropractic Research Archives Collection, and Manual, Alternative, and Natural Therapies Information System databases. A secondary hand search of bibliographies was completed to identify older or nonindexed literature. DATA SELECTION AND EXTRACTION: Articles were identified, which described the characteristics of distraction manipulation beyond a simple description or the results of treatment with distraction manipulation. Data were extracted on the basis of relevance to the stated objective. DATA SYNTHESIS AND RESULTS: Thirty articles were identified. Three were uncontrolled or pilot studies, 3 were basic science studies, and 6 were case series. Most were case reports. Lumbar distraction manipulation is a nonthrust mechanically assisted manual medicine technique with characteristics of manipulation, mobilization, and traction. It is used for a variety of lumbar conditions and chronic pelvic pain. The primary rationale for its use is on the basis of the biomechanical effects of axial spinal distraction. Little data are available describing the in vivo effect of distraction when used in combination with flexion or other motions. CONCLUSIONS: Despite widespread use, the efficacy of distraction manipulation is not well established. Further research is needed to establish the efficacy and safety of distraction manipulation and to explore biomechanical, neurological, and biochemical events that may be altered by this treatment. PMID- 15883581 TI - Unifocal Langerhans cell histiocytosis presenting as an aggressive bone lesion. AB - OBJECTIVE: To discuss a case of unifocal Langerhans cell histiocytosis (eosinophilic granuloma) presenting as an aggressive osseous destructive process. CLINICAL FEATURES: A 6-year-old girl had thigh pain of insidious onset that would wake her at night. Orthopedic testing reproduced local pain, and imaging revealed an osteolytic process in the femoral diaphysis with laminated periosteal reaction. INTERVENTION AND OUTCOME: The patient was referred for further evaluation; unifocal Langerhans cell histiocytosis was found at biopsy and spontaneous healing subsequently reported. CONCLUSIONS: Unifocal Langerhans cell histiocytosis may present with deceptive clinical and imaging findings, of which clinicians should be aware. PMID- 15883582 TI - False negative magnetic resonance imaging results: a report of 2 cases. AB - OBJECTIVES: The purpose of this study is to present 2 clinical case studies in which large herniated disks were not detected on magnetic resonance imaging (MRI), leading to false negative results, and discuss some issues regarding potential shortcomings of MRI. CLINICAL FEATURES: Cases of cervical and lumbar herniated nucleus pulposus (HNP) are presented. Each patient had severe pain and neurological deficit. The patients had positive physical examination findings suggestive of HNP despite the negative MRI scans. Both subsequently underwent myelography and computed tomography to arrive at the final and definitive diagnosis of HNP. INTERVENTION AND OUTCOME: Both patients failed a course of conservative care that included manual and mechanical traction, manual mobilization, myofascial release techniques, epidural steroid injections, oral steroid and narcotic medications, and rehabilitative exercises. Both patients eventually required surgical decompression. CONCLUSION: False negative MRI results in these cases of large HNPs emphasize the importance of case history and physical examination findings as the basis for ordering diagnostic tests. The conclusion of this article is simple; no single diagnostic test should be considered as 100% accurate, as false negative studies may occur that may mislead the patient and clinician. PMID- 15883583 TI - A theoretical model for treatment of soft tissue injuries: treatment of an ankle sprain in a college tennis player. AB - OBJECTIVE: To present theories of manual treatment approaches to soft tissue injuries using an example case report. CLINICAL FEATURES: A college tennis player with an ankle sprain for 6 weeks, not responding to standard treatment, was unable to play tennis or compete in tournaments. INTERVENTION AND OUTCOME: Soft tissue treatment was applied to the ankle for 2 visits. The patient experienced complete resolution of the problem and returned to play without relapse during a 9-month follow-up period. Theories of myofascial distortion treatment are discussed. CONCLUSION: This treatment approach may have potential for soft tissue problems that are not amenable to current therapy approaches. PMID- 15883585 TI - Systemic Agrobacterium tumefaciens-mediated transfection of viral replicons for efficient transient expression in plants. AB - Plant biotechnology relies on two approaches for delivery and expression of heterologous genes in plants: stable genetic transformation and transient expression using viral vectors. Although much faster, the transient route is limited by low infectivity of viral vectors carrying average-sized or large genes. We have developed constructs for the efficient delivery of RNA viral vectors as DNA precursors and show here that Agrobacterium-mediated delivery of these constructs results in gene amplification in all mature leaves of a plant simultaneously (systemic transfection). This process, called "magnifection", can be performed on a large scale and with different plant species. This technology combines advantages of three biological systems (the transfection efficiency of A. tumefaciens, the high expression yield obtained with viral vectors, and the post-translational capabilities of a plant), does not require genetic modification of plants and is faster than other existing methods. PMID- 15883586 TI - Switch from phytotherapy to tamsulosin in patients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia (LUTS/BPH). AB - The aim of this observational prospective study was to evaluate the switch from phytotherapy to tamsulosin 0.4 mg once daily (o.d.) on efficacy, sexual function and tolerability in patients with lower urinary tract symptoms/benign prostatic hyperplasia (LUTS/BPH) who have a poor response to at least 4 weeks of phytotherapy. The switch to tamsulosin 0.4 mg o.d. improves LUTS and related quality of life. Sexual function is also slightly improved. Tamsulosin is as well tolerated as phytotherapy and abnormal ejaculation appears to be no problem. Tamsulosin is perceived by both patients and urologists to be superior to preceding phytotherapy. PMID- 15883587 TI - Common VKORC1 and GGCX polymorphisms associated with warfarin dose. AB - We report a novel combination of factors that explains almost 60% of variable response to warfarin. Warfarin is a widely used anticoagulant, which acts through interference with vitamin K epoxide reductase that is encoded by VKORC1. In the next step of the vitamin K cycle, gamma-glutamyl carboxylase encoded by GGCX uses reduced vitamin K to activate clotting factors. We genotyped 201 warfarin-treated patients for common polymorphisms in VKORC1 and GGCX. All the five VKORC1 single nucleotide polymorphisms covary significantly with warfarin dose, and explain 29 30% of variance in dose. Thus, VKORC1 has a larger impact than cytochrome P450 2C9, which explains 12% of variance in dose. In addition, one GGCX SNP showed a small but significant effect on warfarin dose. Incorrect dosage, especially during the initial phase of treatment, carries a high risk of either severe bleeding or failure to prevent thromboembolism. Genotype-based dose predictions may in future enable personalised drug treatment from the start of warfarin therapy. PMID- 15883588 TI - Stochasticity in gene expression: from theories to phenotypes. AB - Genetically identical cells exposed to the same environmental conditions can show significant variation in molecular content and marked differences in phenotypic characteristics. This variability is linked to stochasticity in gene expression, which is generally viewed as having detrimental effects on cellular function with potential implications for disease. However, stochasticity in gene expression can also be advantageous. It can provide the flexibility needed by cells to adapt to fluctuating environments or respond to sudden stresses, and a mechanism by which population heterogeneity can be established during cellular differentiation and development. PMID- 15883589 TI - Whole-genome patenting. AB - Gene patenting is now a familiar commercial practice, but there is little awareness that several patents claim ownership of the complete genome sequence of a prokaryote or virus. When these patents are analysed and compared to those for other biological entities, it becomes clear that genome patents seek to exploit the genome as an information base and are part of a broader shift towards intangible intellectual property in genomics. PMID- 15883590 TI - The effects of genetic factors on selected indicators of the activity of the sympathoadrenal system and the renin-angiotensin-aldosterone system in twins. AB - BACKGROUND: There are numerous data indicating a significant role of the sympathoadrenal system and the reninangiotensin- aldosterone system in the regulation of blood pressure and the pathogenesis of essential hypertension. However, the genetic background of essential hypertension remains unclear. AIM: To determine the effects of genetic factors on selected indicators of the activity of the sympathoadrenal system and the renin-angiotensin-aldosterone system in twins. METHODS: We studied 39 monozygotic twin pairs (age 33+/-7 years) and 37 same-gender dizygotic twin pairs (age 36+/-7 years). We measured blood and urine adrenaline (A), noradrenaline (NA), dopamine (DA) and aldosterone (ALD) levels, as well as plasma renin activity (PRA) and serum angiotensin-converting enzyme (ACE) activity. Parameters of the genetic models for age- and gender adjusted data were estimated by model fitting and path analysis technique using LISREL 8. RESULTS: The effects of genetic factors on the variability of blood and urine catecholamine levels were 69% and 65% for A, 42% and 76% for NA, and 58% and 40% for DA, respectively. We also found shared environmental components for blood NA (28%) and urine DA (17%). Genetic factors accounted for 36% of the variability of PRA and 80% of the variability of ACE. ALD levels were related only to environmental factors (including a shared environmental component, estimated at 25%, for urine ALD). CONCLUSIONS: We found significant effects of genetic factors on the activity of the sympathoadrenal system, as indicated by blood and urine catecholamine levels. We also found the effect of genetic factors on PRA and ACE, but not on aldosterone levels. PMID- 15883591 TI - Differences between cardiologists and internists in the management of heart failure. Medical guidelines compared with every-day practice. AB - BACKGROUND: A marked progress in the treatment of chronic heart failure (CHF) took place during the last decade. Large, randomised studies documented the role of angiotensin-converting enzyme inhibitors (ACE-I) and beta-blockers in the reduction of mortality and morbidity in CHF. AIM: To assess differences in the management of CHF patients between cardiologists and internists. METHODS: Medical records of 433 patients with CHF, aged 38-98 years, hospitalised between October 2000 and May 2002 in an academic centre, were retrospectively analysed. Cardiologists treated 241 patients, and internists - 192 patients. In addition, 12 randomly selected physicians who treated CHF patients, filled-in a questionnaire concerning CHF treatment. Next, the answers were compared with the actual treatment, documented in patients medical files. RESULTS: There were significant differences in the CHF treatment between cardiologists and internists. Cardiologists significantly more often prescribed ACE-I in NYHA class III patients (77% vs 58%, p=0.003), beta-blockers in all NYHA classes (80% vs 57%, p<0.001), loop diuretics in NYHA classes III and IV (76% vs 42%, p<0.001 and 91% vs 44%, p=0.005), and thiazides in NYHA class III patients (36% vs 11%, p<0.001). Internists more often used loop diuretics (37% vs 15%, p<0.001), digoxin (28% vs 7%, p<0.001) and aldosterone antagonists (24% vs 13%, p=0.022) in NYHA class II patients. All cardiologists and almost half of internists declared combined usage of ACE-I and beta-blockers at increasing dosages as the treatment was continued. The highest concordance between declared and actual medication concerned the concomitant use of ACE-I and beta-blockers. CONCLUSIONS: Cardiologists used more often ACE-I and beta-blockers than internists in the treatment of CHF patients. A satisfactory knowledge of treatment guidelines was not associated with widespread usage of ACE-I and beta-blockers in every-day practice. PMID- 15883592 TI - Prognostic value of exercise-induced QT dispersion in patients after acute myocardial infarction. AB - BACKGROUND: Electrocardiographic exercise tests are widely recommended for patients before discharge after myocardial infarction, what justify the search for new variables which may improve their prognostic value. QT dispersion in 12 lead ECG reflects the heterogeneity of ventricular repolarisation. Increased QT dispersion is a noninvasive marker of ischaemia and electrical instability. AIM: Evaluation of the prognostic value of exercise-induced changes of QT dispersion in patients after an acute myocardial infarction. METHODS: Heart rate limited treadmill exercise test according to modified Bruce was performed 14+/-5 days after infarction in 77 patients (age 56+/-11,8 female). QT dispersion was measured at rest and on peak exercise. Patients were followed up for mean 88 months. RESULTS: QT dispersion was higher at peak exercise in those patients who died due to cardiovascular causes (n=8) or suffered from non-fatal myocardial infarction during follow-up (n=15), than in remaining group (71+/-20 vs 58+/-22 msec, p<0.01). At rest QT dispersion was similar in both groups (64+/-17 vs 66+/ 20 msec, NS). CONCLUSIONS: The lack of an exercise-induced decrease in QT dispersion identifies a subgroup of patients after myocardial infarction with a poor long-term prognosis. PMID- 15883593 TI - Diagnostic value of BNP in suspected perimyocarditis--a preliminary report. AB - BACKGROUND: Diagnosis of perimyocarditis is often very challenging. Clinical presentation includes chest pain and ECG changes which are difficult to interpret. Clinical course is usually mild, however, some patients develop heart failure symptoms and require aggressive treatment. Plasma b-type natriuretic peptide (BNP) is a marker of the hemodynamical impairment of the heart. Its diagnostic role in patients with acute perimyocarditis has not yet been examined. AIM: To assess the usefulness of BNP measurement in the diagnosis of perimyocarditis. METHODS: The study group consisted of 14 consecutive patients (13 males, mean age 32.1+/-12.4 years) with suspected perimyocarditis (history of influenza, typical symptoms, ECG and echocardiographic results as well as myocardial necrotic markers). Plasma BNP was assessed at bedside at the time of admission. RESULTS: Plasma BNP, measured in 12 patients, was 163+/-154 pg/mL (max. 519 pg/mL) and exceeded upper normal level in 6 (50%) patients. When normal levels were adjusted for age and gender, 9 (80%) patients had elevated BNP. One patient had heart failure symptoms and a BNP level of 205 pg/mL. In all 4 patients who had transient myocardial contractility disturbances, detected by echocardiography, BNP level exceeded 100 pg/mL. CONCLUSIONS: BNP level is increased in some patients with acute perimyocarditis. BNP elevation is probably associated with hemodynamical stress caused by transient contractility abnormalities. Diagnostic and prognostic role of BNP in acute perimyocarditis requires further studies. PMID- 15883594 TI - Nuclear magnetic resonance for the assessment of the effects of alcohol septal ablation in hypertrophic cardiomyopathy. AB - Alcohol ablation of the interventricular septum is a well-established method of treatment in hypertrophic obstructive cardiomyopathy (HOCM). The extent of ablation-induced iatrogenic infarction is usually assessed by the use of echocardiography. The role of nuclear magnetic resonance (NMR) in this setting has not yet been well established. In this report we describe five patients with HOCM who underwent alcohol septal ablation and subsequent NMR evaluation. The usefulness of NMR in these patients is discussed and comparison with echocardiographic results is presented. PMID- 15883595 TI - [Cardiac revascularisation in a high-risk patient--case report]. AB - A case of a patient with left main stem stenosis and significant stenosis in the other coronary arteries is presented. The patient was disqualified from coronary artery bypass grafting (CABG) because of many surgical risk factors and underwent successful coronary angioplasty. Three months later the patient returned to the hospital because of unstable angina. Control coronary angiography revealed in stent restenosis. The patient underwent CABG and drug-eluting stent implantation with good clinical result. PMID- 15883596 TI - [Acute aortic dissection in a patient with advanced coronary artery disease--a case report]. AB - A case of a 58-year-old male with acute aortic dissection is presented. A few months earlier the patient underwent transmyocardial laser revascularisation and implantation of venous graft to the left anterior descending coronary artery due to three-vessel disease. Acute aortic dissection was successfully treated by surgery. Therapeutic options in patients with acute aortic dissection and advanced coronary artery disease are discussed. PMID- 15883597 TI - [The effects of intracoronary brachytherapy on long-term patency of previously chronically occluded bypass graft--a case report]. AB - A case of a 70-year-old female who underwent CABG (four grafts) 12 years earlier, is presented. Following surgery, the patient suffered from two reinfarctions. Coronary angiography revealed total occlusion of the graft implanted to the right coronary artery which was treated by coronary angioplasty. Because of restenosis, the patient underwent successful intracoronary brachytherapy. PMID- 15883598 TI - [Standard pharmacological therapy associated with percutaneous coronary interventions in patients with stable angina]. PMID- 15883607 TI - [Cor triatriatum in an adult patient--an echocardiographic case report]. PMID- 15883608 TI - [Electrocardiography in left main coronary artery stenosis]. PMID- 15883609 TI - [Angiogram of the month]. PMID- 15883610 TI - [Endocardial and epicardial mapping during RF ablation of left ventricular ectopy]. PMID- 15883611 TI - [Blood pressure reduction in hypertension--what is the optimal target value?]. PMID- 15883612 TI - Adalimumab therapy: clinical findings and implications for integration into clinical guidelines for rheumatoid arthritis. AB - Adalimumab (Humira) is the first fully human monoclonal anti-tumor necrosis factor (TNF) antibody available. Similar to the other TNF-alpha blockers, adalimumab has been shown to effectively reduce the symptoms and signs of rheumatoid arthritis and prevent the progression of erosive joint changes seen on radiological examination, which would lead to disabling joint damage. Clinical guidelines recommend the use of TNF blockers, specifically etanercept and infliximab (the only two available when the guidelines were issued) as treatment options for adults with rheumatoid arthritis who continue to have clinically active disease that has not responded adequately to two conventional disease modifying antirheumatic drugs (DMARDs). The clinical results available using adalimumab, and summarized in this review, reveal a clinical profile similar to etanercept and infliximab, achieving similarly high response rates, suppression of joint damage, and improvements in quality of life and disability, together with a good safety profile. Being a fully human monoclonal antibody, adalimumab may induce less antigenicity than these other agents, which might also be advantageous in maintaining the level of effectiveness. However, direct comparisons in controlled, long-term trials are needed to draw conclusions about which agent to try first in the sequence of DMARDs considered for patients. PMID- 15883613 TI - Pexelizumab: a novel therapy for myocardial ischemia-reperfusion. AB - Despite significant advances in the study of myocardial reperfusion, patients with epicardial coronary reperfusion still have adverse clinical outcomes. This is in part due to an inflammatory reaction in the injured tissue. Inflamed myocardial tissue after ischemia and reperfusion releases several cytokines and toxic metabolites that lead to reperfusion injury and cellular apoptosis. Complement activation appears to mediate myocardial damage through both these pathways. Recently, pexelizumab, a novel C5 complement monoclonal antibody fragment, has been developed to prevent the complement-mediated myocardial damage from myocardial ischemia and reperfusion. In animal studies, pexelizumab decreased the amount of myocardial damage with ischemia and reperfusion. Pexelizumab has been studied in phase II and phase III clinical trials involving patients undergoing coronary artery bypass grafting and in patients with acute myocardial infarction as an adjunct to reperfusion therapy. Two large clinical trials are currently underway in coronary artery bypass surgery and acute myocardial infarction patients undergoing primary percutaneous revascularization. This review will cover the pathophysiological role of complement activation with regards to ischemia-reperfusion injury, the novel compound pexelizumab, and both the preclinical and clinical data for pexelizumab use in coronary artery bypass surgery and acute myocardial infarction patients. PMID- 15883614 TI - Inflammatory biomarkers and statins. AB - The concept of atherosclerosis as an inflammatory disorder has led to the exploration of new pathogeneses of this disease. In this regard, the levels of several inflammatory molecules are frequently increased in subjects at high risk of developing an acute coronary event. With a simple analysis we can characterize the circulating levels of a marker and its therapeutic modulation with various drugs. In this review we have analyzed different inflammatory markers currently used, such as C-reactive protein (CRP), CD40 ligand, adhesion molecules and chemokines, and their possible modulation by therapeutic intervention with 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors. Moreover, in the future, new technologies will allow us to discover new markers, or sets of them, that could indicate the direction to be taken in the prevention and treatment of cardiovascular diseases. PMID- 15883615 TI - Prostate biopsy in the diagnosis of prostate cancer: current trends and techniques. AB - The advent of prostate specific antigen (PSA) screening and transrectal ultrasonography (TRUS) has had a significant impact on the detection of prostate cancer over the last 15 years. The mean age at diagnosis has decreased and the most common stage at diagnosis is now localized disease. TRUS guidance, spring loaded biopsy needles, utilization of oral antibiotic prophylaxis, developments in local anesthesia, increases in the number of cores sampled and the use of site specific containers have all made the prostate biopsy easier to perform and more accurate. The indications for an initial prostate biopsy have been strongly influenced by digital rectal examinations (DREs), PSA levels and the PSA-related parameters of velocity, density, and percent free. These parameters, along with abnormal histology, also dictate the need for a repeat biopsy. With the better, earlier, and more patient-friendly usage of the prostate biopsy, there has been a decrease in the mortality rate of prostate cancer. PMID- 15883616 TI - The genetics of susceptibility to cutaneous melanoma. AB - The most common genetic determinants of skin cancer are the genes that control skin color so that the genes expressed as black skin are protective. Within the white population variants at the MC1R locus are shown to increase susceptibility, and recent evidence suggests that polymorphisms in the OCA2 gene interact to modify risk. Other as-yet-unidentified pigment genes may also play a role in susceptibility to melanoma. Thus MC1R variants are the most common low-penetrance melanoma-susceptibility genes so far identified. Other putative low-penetrance susceptibility genes have been explored using candidate gene approaches. Good candidates such as the DNA repair gene XRCC3 and polymorphisms of EGF have been studied but excluded. Other candidates such as BRAF polymorphisms and variants at the CDKN2A locus remain to be fully investigated. More progress has been made in identifying high-penetrance genes, however. The most common (and probably the most penetrant) susceptibility locus is the CDKN2A locus coding for two tumor suppressor proteins, p16 and p14ARF. Mutations at this locus that impact p16, p14ARF or both proteins all increase susceptibility to melanoma. Families inheriting such mutants are at increased risk of cutaneous melanoma, and the penetrance is increased by residence in sunny climates and co-inheritance of MC1R variants. Some families also appear to be at increased risk of pancreatic cancer, but the determinants of susceptibility to pancreatic cancer are not yet understood. Very rare families have germline mutations in the CDK4 gene which impact on the p16 binding site. There are other high-penetrance susceptibility genes, however, which remain undiscovered. There is evidence of one at 1p22. The Melanoma Genetics Consortium (www.genomel.org) continues to explore this and the genetic epidemiology of the CDKN2A locus. PMID- 15883617 TI - The genetics of osteoporosis. AB - Osteoporosis is a common health problem among the elderly, especially postmenopausal women. It is characterized by fragile bones susceptible to low trauma fractures. Osteoporosis is a complex disease determined by genetic and environmental factors, as well as the possible interactions among these factors. Twin and family studies have shown that genetic factors play an important role in osteoporosis. Numerous genetic studies have been performed to hunt for genes underlying the disease risk. In this review, we briefly summarize and discuss the current status of knowledge about osteoporosis, with special emphasis on the progress achieved recently in: 1) heritability and choice of osteoporosis study phenotypes; 2) the approaches for gene mapping and identification for osteoporosis; 3) the candidate gene association studies; 4) the linkage mapping studies for osteoporosis; 5) some potential explanations for the inconsistent results; and 6) the pharmacogenetic studies for osteoporosis. PMID- 15883618 TI - P2X and P2Y receptors as possible targets of therapeutic manipulations in CNS illnesses. AB - Adenine and/or uridine nucleotide-sensitive receptors are classified into two types belonging to the ligand-gated ionotropic family (P2X) and the metabotropic, G-protein-coupled family (P2Y). In humans, seven different P2X receptors (P2X(1 7)) and eight different P2Y receptors (P2Y(1), P2Y(2), P2Y(4), P2Y(6), P2Y(11 14)) have been detected hitherto. All P2 receptors are expressed in the CNS, with the preferential expression of the P2X(2), P2X(4), P2X(6) and P2Y(1) receptors in neurons. In addition to the neurotransmitter and modulator functions, neurite outgrowth, proliferation of glial cells and the expression of transmitter receptors at target cells have also been suggested to be regulated by extracellular nucleotides in the nervous system. In spite of the expanding knowledge in the purinergic research field, the present therapeutic utilization of P2 receptor ligands is mostly related to peripheral diseases such as thromboembolic disorders and cystic fibrosis. In this review we provide some evidence that P2 receptors play an important role in the regulation of CNS functions related to hippocampal activity, the mesolimbic dopaminergic system and the nociceptive system. The role of purinergic receptors located on astrocytes/microglia and implications of these receptors for neurodegenerative/neuroinflammatory disorders, CNS injury and epilepsy will be highlighted as well. PMID- 15883619 TI - TIRC7 pathway as a target for preventing allograft rejection. AB - A number of leukocyte surface molecules play an essential role during immune activation. Targeting of these molecules utilizing antibodies serves as a specific therapeutic approach for the treatment of a variety of human diseases. Antibodies targeting a number of leukocyte surface molecules were shown to induce tolerance to transplants in several animal models. A novel membrane molecule, T cell immune response cDNA 7 (TIRC7), has been shown to be an essential protein in the regulation of lymphocyte activation. TIRC7 does not share any homology with other known membrane proteins expressed during the course of lymphocyte activation and does not belong to any of the known costimulatory, cytokine, chemokine or receptor families. TIRC7, a highly conserved protein across species, is expressed in immune tissues such as spleen, lymph nodes, and T and B lymphocytes. Antibodies against extracellular domains of TIRC7 prolong allograft survival in rat and mouse transplantation models. The prevention of rejection is mediated at least partially via induction of cytotoxic T lymphocyte antigen 4 (CTLA4) in T cells. Functional cellular assays utilizing TIRC7-deficient mice splenocytes show that TIRC7 does have an impact not only on T-cell, but also on B cell response. Subtherapeutic amounts of FK506 and anti-TIRC7 monoclonal antibody prolong graft survival, suggesting synergistic effects with calcineurin inhibitors. Targeting TIRC7 with monoclonal antibody might serve as a promising therapeutic strategy for preventing allograft rejection in humans and treatment of other immune-related diseases. Acutely rejected human kidney allografts show strong expression of TIRC7 despite treatment with calcineurin inhibitors. Therefore, monitoring TIRC7 expression may facilitate an early diagnostic tool of acute rejection. TIRC7 seems to belong to a group of targets with dual roles in disease pathogenesis, so-called theranostics, which can be utilized to treat and diagnose diseases. PMID- 15883620 TI - Trends in exploration of therapeutic targets. AB - Lead discovery against a preselected therapeutic target is a key component in modern drug development. Continuous effort and increasing interest has been directed at the search for new targets, which has led to the identification of a growing number of them. Data from the therapeutic target database, at http://bidd.nus.edu.sg/group/cjttd/ttd.asp, show that, as of July 2004, the number of documented targets of marketed and investigational drugs has reached 1,174 distinct proteins (including subtypes) and 27 nucleic acids, 239 of which are targets of the marketed drugs. Analysis of these targets, particularly those of recently approved drugs and patented investigational agents, provide useful hints about general trends of target exploration and current focus in drug discovery for the treatment of high impact diseases needing effective or more treatment options. PMID- 15883621 TI - Current development of lentiviral-mediated gene transfer. AB - Lentiviral-based vectors have been widely used lately in preclinical studies and for the in vitro genetic manipulation of embryonic and adult stem cells. They allow for the transduction of nondividing cells and for stable gene expression. On these grounds, lentiviral vectors look promising for eventual applications for the gene-based treatment of neurological disorders, cardiopathies and inherited or acquired genetic diseases. The best developed and characterized lentiviral vector system is based on the human immunodeficiency virus type 1 (HIV-1). However, safety concerns preclude any possible application of HIV-1-derived vectors in clinical trials. For this reason, gene therapists are currently improving the design of other lentiviral vectors, such as feline immunodeficiency virus, equine anemia infectious virus and Visna virus. These lentiviruses are not pathogenic in humans and are only distantly related to primate Retroviridae. This review summarizes the achievements in improving the design of lentiviral vector systems that are not based on HIV-1. PMID- 15883622 TI - Molecule of the month. MPC-7869 (Flurizan). PMID- 15883623 TI - Trends in medicinal chemistry 2004. AB - On December 2, 2004, the Society for Medicines Research held the seventh Trends in Medicinal Chemistry one-day meeting. The meeting brought together speakers from Europe representing both academia and industry and provided an overview of some of the latest approaches being taken in a range of therapeutic areas such as oncology, antiinfectives, CNS disease and reproductive medicine. PMID- 15883624 TI - New targets and new drugs down under in 2004. AB - The Second Australian Health and Medical Research Congress, held November 21-26, 2004, in Sydney, Australia, brought together 28 specialist societies across a broad range of scientific endeavor. The topics of the plenary lectures included signal transduction by stress-activated mitogen-activated protein kinases, whether human cancer can be treated before it is symptomatic, the modeling of human cardio-myopathies via genetic manipulation in animal models, and biological individuality and the response to regular exercise. The symposia included translating basic research to clinical practice, molecular mechanisms and anticancer therapies, modulating signaling pathways to treat/prevent disease, therapeutic frontiers in bone and joint disease, pleiotropic effects of statins in the cardiovascular system, and angiogenesis inhibitors in cancer and atherosclerosis therapy. PMID- 15883627 TI - Potential impact of osteoporosis treatment on hip fracture trends. PMID- 15883628 TI - Population trends in BMD testing, treatment, and hip and wrist fracture rates: are the hip fracture projections wrong? AB - A worldwide epidemic of hip fractures has been predicted. Time trends in BMD testing, bone-sparing medications and hip and wrist fractures in the province of Ontario, Canada, were examined. From 1996 to 2001, BMD testing and use of bone sparing medications increased each year, whereas despite the aging of the population, wrist and hip fracture rates decreased. INTRODUCTION: If patients with osteoporosis are being diagnosed and effective treatments used with increasing frequency in the population, rates of hip and wrist fractures will remain stable or possibly decrease. We report here time trends in BMD testing, prescriptions for bone-sparing medications, hip and wrist fracture rates, and population projections of fracture rates to 2005 in the province of Ontario, Canada. MATERIALS AND METHODS: Ontario residents have universal access to Medicare. To examine time trends in BMD testing, all physician claims for DXA from 1992 to 2001 were selected from the Ontario Health Insurance Plan (OHIP) database. Trends in prescribing were examined from 1996 to 2003 using data from the Ontario Drug Benefit plan, which provides coverage to persons > or = 65 years of age. Actual numbers of hip and wrist fractures were determined for 1992-2000 and population projections for 2001-2005 using time-series analysis. Wrist fractures were identified in the OHIP database and hip fractures through hospital discharge abstracts. RESULTS: From 1992 to 2001, the number of BMD tests increased 10-fold. There has been a steady increase in the number of persons filling prescriptions for antiresorptives (12,298 in 1996 to 225,580 in 2003) and the majority were for etidronate. For women, the rate of decline for wrist fractures is greater than that for hip fractures. The rate of hip fracture was fairly constant around 41 per 10,000 women > or = 50 years between 1992 and 1996. In 1997, the hip fracture rate began to decrease, and the population projections suggest that this downward trend will continue to a rate of 33.1 per 10,000 in 2005. CONCLUSIONS: Our findings suggest that fracture rates may be on the decline, despite the aging of the population, because of increased patterns of diagnosis and treatment for osteoporosis. PMID- 15883629 TI - Reduced training is associated with increased loss of BMD. AB - This 8-year controlled, follow-up study in 66 Swedish soccer women evaluated the effect of training and reduced training on BMD. The players who retired during the follow-up lost BMD in the femoral neck, whereas the controls did not. INTRODUCTION: Physical activity during adolescence increases BMD, but whether the benefits are retained with reduced activity is controversial. MATERIALS AND METHODS: At baseline, DXA evaluated BMD in 48 active female soccer players with a mean age of 18.2 +/- 4.4 (SD) years, in 18 former female soccer players with a mean age of 43.2 +/- 6.2 years and retired for a mean of 9.4 +/- 5.3 years, and in 64 age- and sex-matched controls. The soccer women were remeasured after a mean of 8.0 +/- 0.3 years, when 35 of the players active at baseline had been retired for a mean of 5.3 +/- 1.6 years. RESULTS AND CONCLUSIONS: The players still active at follow-up had a higher BMD at baseline than the matched controls in the femoral neck (FN; 1.13 +/- 0.19 versus 1.00 +/- 0.13 g/cm2; p = 0.02). The yearly gain in BMD during follow-up was higher in the active players than in the controls in the leg (0.015 +/- 0.006 versus 0.007 +/- 0.012 g/cm2, p = 0.04). The soccer players who retired during follow-up had a higher BMD at baseline than the matched controls in the FN (1.13 +/- 0.13 versus 1.04 +/- 0.13 g/cm2; p = 0.005). The players that retired during follow-up lost BMD, whereas the controls gained BMD during the study period in the FN (-0.007 +/- 0.01 versus 0.003 +/- 0.02 g/cm2 yearly; p = 0.01). The soccer players already retired at baseline had higher BMD at study start than the matched controls in the leg (1.26 +/- 0.09 versus 1.18 +/- 0.10 g/cm2; p = 0.01). The former players who were retired at study start lost BMD, whereas the controls gained BMD during the study period in the trochanter (-0.006 +/- 0.01 versus 0.004 +/- 0.014 g/cm2 yearly; p = 0.01). This study shows that, in girls, intense exercise after puberty is associated with higher accrual of BMD, and decreased physical activity in both the short term and long-term perspective is associated with higher BMD loss than in controls. PMID- 15883630 TI - Serum retinoids and beta-carotene as predictors of hip and other fractures in elderly women. AB - There is debate about the possible deleterious effect of excessive vitamin A exposure on fracture risk. In this nested case control study in older women (312 cases and 934 controls), serum retinol, retinyl palmitate, and beta-carotene were not associated with fracture risk, and there was no evidence of excess risk with multivitamin or cod liver oil supplementation. INTRODUCTION: Recent studies have suggested that higher vitamin A intake may account for a component of fracture risk within the general population and that supplemental vitamin A may be harmful even within recommended limits. No studies have examined the relationship between biochemical retinol status and fracture in older women. MATERIALS AND METHODS: We examined serum retinol, retinyl palmitate, and beta-carotene as predictors of incident hip and other fractures in a large prospective study of British women over the age of 75 years (n = 2606, 312 incident osteoporotic fractures, 92 incident hip fractures; mean follow-up duration, 3.7 years). Fasting blood samples (9:00-11:00 a.m.) were collected at baseline. Using a case-control design (three controls per case), serum retinol, retinyl palmitate, and beta-carotene were assessed as univariate predictors of incident osteoporotic fracture or hip fracture. Baseline BMD at the total hip, age, 25(OH)D, serum beta Crosslaps, bone specific alkaline phosphatase, weight, height, and smoking were considered as covariates in a multivariate model. RESULTS: Serum retinol, retinyl palmitate, and beta-carotene were not significant univariate predictors of either hip fracture or any fracture (all p > 0.05; Cox proportional hazards regression). For all osteoporotic fractures, the hazard ratio (HR) was 0.92 (95% CI, 0.81-1.05) per 1 SD increase in serum retinol. Risk of any osteoporotic fracture was slightly less in the highest quartile of serum retinol compared with the lowest quartile (HR, 0.85; 95% CI, 0.69-1.05; p = 0.132) There was a tendency for increased serum retinol to predict benefit rather than harm in terms of BMD (r = 0.09, p = 0.002). Multivitamin or cod liver oil supplementation was associated with a significantly lower risk of any fracture (HR, 0.76; 95% CI, 0.60-0.96; p = 0.021). In multivariate analysis, only age, total hip BMD, and weight were associated with fracture risk (p < 0.05). CONCLUSIONS: We found no evidence to support any skeletal harm associated with increased serum indices of retinol exposure or modest retinol supplementation in this population. PMID- 15883631 TI - Homocysteine and vitamin B12 status relate to bone turnover markers, broadband ultrasound attenuation, and fractures in healthy elderly people. AB - Hyperhomocysteinemia may contribute to the development of osteoporosis. The relationship of Hcy and vitamin B12 with bone turnover markers, BUA, and fracture incidence was studied in 1267 subjects of the Longitudinal Aging Study Amsterdam. High Hcy and low vitamin B12 concentrations were significantly associated with low BUA, high markers of bone turnover, and increased fracture risk. INTRODUCTION: Hyperhomocysteinemia may contribute to the development of osteoporosis. Vitamin B12 is closely correlated to homocysteine (Hcy). The main objective of our study was to examine the association of Hcy and vitamin B12 status and the combined effect of these two with broadband ultrasound attenuation (BUA), bone turnover markers, and fracture. MATERIALS AND METHODS: Subjects were 615 men and 652 women with a mean age of 76 +/- 6.6 (SD) years of the Longitudinal Aging Study Amsterdam (LASA). At baseline (1995/1996), blood samples were taken after an overnight fast for dairy products. Plasma Hcy was measured with IMx, serum vitamin B12 with competitive immunoassay (IA) luminescence, serum osteocalcin (OC) with immunoradiometric assay (IRMA), and urinary excretion of deoxypyridinoline (DPD) with competitive IA and corrected for creatinine (Cr) concentration. CVs were 4%, 5%, 8%, and 5%, respectively. BUA was assessed in the heel bone twice in both the right and left calcaneus. Mean BUA value was calculated from these four measurements. CV was 3.4%. After baseline measurements in 1995, a 3-year prospective follow-up of fractures was carried out until 1998/1999. Subjects were grouped by using two different approaches on the basis of their vitamin B12 concentration, normal versus low (<200 pM) or lowest quartile (Q1) versus normal quartiles (Q2-Q4), and Hcy concentration, normal versus high (>15 microM) or highest quartile (Q4) versus normal quartiles (Q1 Q3). Analysis of covariance was performed to calculate mean values of BUA, OC, and DPD/Cr(urine) based on the specified categories of Hcy and vitamin B12 and adjusted for several confounders (potential confounders were age, sex, body weight, body height, current smoking [yes/no], mobility, cognition). The relative risk (RR) of any fracture was assessed with Cox regression analysis. Quartiles were used when Hcy and vitamin B12 were separately studied in their relationship with fracture incidence. RESULTS: Fourteen percent of the men and 9% of the women had high Hcy (>15 microM) and low vitamin B12 (<200 pM) concentrations. Women with vitamin B12 levels <200 pM and Hcy concentrations >15 microM had lower BUA, higher DPD/Cr, and higher OC concentrations than their counterparts. In men, no differences were found between the different Hcy and vitamin B12 categories in adjusted means of BUA, OC, or DPD/Cr(urine). Twenty-eight men and 43 women sustained a fracture during the 3-year follow-up period. The adjusted RR for fractures (95% CI) for men with high Hcy and/or low vitamin B12 concentrations was 3.8 (1.2-11.6) compared with men with normal Hcy and vitamin B12 concentrations. Women with high Hcy and/or low vitamin B12 concentrations had an adjusted RR for fractures of 2.8 (1.3-5.7). CONCLUSIONS: High Hcy and low vitamin B12 concentrations were significantly associated with low BUA, high markers of bone turnover, and increased fracture risk. PMID- 15883632 TI - Human bone collagen synthesis is a rapid, nutritionally modulated process. AB - We developed a direct assay of human bone collagen synthesis using [13C] or [15N] proline and applied it to determine the effects of feeding in young healthy men. Surprisingly, postabsorptive bone collagen synthesis is not sluggish, being approximately 0.07%/h more rapid than that of muscle protein, and capable of being stimulated within 4 h of intravenous feeding by 66 +/- 13%. INTRODUCTION: All current methods for estimation of bone collagen turnover are indirect, depending on the assay of collagen "markers." Our aim was to develop a direct method for human bone collagen synthesis to be used to study its physiology and pathology, and specifically, in the first instance, the effect of feeding. MATERIALS AND METHODS: We applied, over 2 h, flooding doses of [13C] and [15N] proline to label iliac crest bone collagen in eight young healthy men. The rate of collagen synthesis was determined as the rate of labeling of collagen hydroxyproline (assayed by gas chromatography-combustion-isotope ratio mass spectrometry in collagen extracted by differential solubility) compared with plasma proline labeling (assayed by gas chromatography-mass spectrometry). We also determined (in a second group of eight young healthy men) the effect of intravenous nutrition (glucose, lipid emulsion, and amino acids (in the ratio of 55%:30%:15% energy, respectively). RESULTS: Free bone proline labeling was 92 +/- 6% of that of plasma proline, supporting the flooding dose assumption. Human iliac crest bone collagen is heterogeneous, with NaCl-EDTA, 0.5 M acetic acid, pepsin-acetic acid, and hot water-extractable pools being responsible for approximately 1%, 3%, 8%, and 81% of content, respectively. The synthetic rates were 0.58 +/- 0.1, 0.24 +/- 0.05, 0.07 +/- 0.02, and 0.06 +/- 0.01%/h, respectively, giving an average rate of approximately 0.066%/h. [13C] and [15N] proline gave identical results. Intravenous nutrition caused the disappearance of proline label from the procollagen pool and its increased appearance in the less extractable pools, suggesting nutritional stimulation of collagen processing. CONCLUSION: The results show (1) that iliac crest bone collagen synthesis is faster than generally assumed and of the same order as muscle protein turnover and (2) that feeding increases synthesis by approximately 66%. Given its ability to detect physiologically meaningful responses, the method should provide a new approach to studying the regulation of bone collagen turnover. PMID- 15883633 TI - A frequent regulatory variant of the estrogen-related receptor alpha gene associated with BMD in French-Canadian premenopausal women. AB - Genes are important BMD determinants. We studied the association of an ESRRA gene functional variant with BMD in 1335 premenopausal women. The ESRRA genotype was an independent predictor of L2-L4 BMD, with an effect similar to smoking and equivalent to a 10-kg difference in weight. INTRODUCTION: Several genetic polymorphisms have been associated with osteoporosis or osteoporosis fractures, but no functional effect has been shown for most of these gene variants. Because functional studies have implicated estrogen-related receptor alpha (ESRRA) in bone metabolism, we evaluated whether a recently described regulatory variant of the ESRRA gene is associated with lumbar and hip BMD as measured by DXA and with heel bone parameters as measured by quantitative ultrasound (QUS). MATERIALS AND METHODS: Heel bone parameters were measured by right calcaneal QUS in 1335 healthy French-Canadian premenopausal women, and one-half of these women also had their BMD evaluated at two sites: femoral neck and lumbar spine (L2-L4) by DXA. All bone measures were tested separately for association with the ESRRA genotype by analysis of covariance. The significance of the ESRRA contribution to the model was also assessed by two different permutation tests. RESULTS: A statistically significant association between ESRRA genotype and lumbar spine BMD was observed: women carrying the long ESRRA genotype had a 3.9% (0.045 g/cm2) higher lumbar spine BMD than those carrying the short ESRRA genotype (p = 0.004), independently of other risk factors measured. This effect of ESRRA genotype is similar to the effect of smoking and equivalent to a 10-kg difference in weight. This association was confirmed by permutation tests (p = 0.004). The same trend was observed for femoral neck BMD (2.6%, p = 0.07). However, no association was observed between ESRRA and QUS heel bone measures. CONCLUSION: These results support the genetic influence of this ESRRA regulatory variant on BMD. PMID- 15883634 TI - Vitamin D receptor Fok1 polymorphisms affect calcium absorption, kinetics, and bone mineralization rates during puberty. AB - Few studies of the VDR polymorphisms have looked at calcium metabolism or long term effects. We measured bone mineralization and calcium metabolic parameters longitudinally in a group of 99 adolescents. We found a significant relationship between calcium absorption and skeletal calcium accretion and the Fok1, but not other VDR or related, genetic polymorphisms. It seems that the Fok1 polymorphism directly affects bone mineralization during pubertal growth through an effect on calcium absorption. INTRODUCTION: There are few data regarding the relationship between genetic markers for low bone mass and changes in calcium metabolism in childhood or adolescence. We sought to identify the effects of polymorphisms of the vitamin D receptor (VDR) on calcium and bone mineral metabolism in a longitudinal study of pubertal adolescents. MATERIALS AND METHODS: Adolescents (n = 99) received comprehensive stable isotope studies of calcium absorption, bone calcium kinetics, and bone mineralization. Studies were repeated 12 months later. Polymorphisms of putative genetic markers were determined and related to bone mineralization and calcium metabolic finding. Results were analyzed by ANOVA in which changes over time were determined using the initial value as a covariate. RESULTS: Polymorphisms of the Fok1 gene of the VDR were significantly related to calcium absorption (p = 0.008) and whole body BMC (p = 0.03) and BMD (p = 0.006). The Fok1 effect on whole body BMD was significant for those with Ca intake >800 mg/day (p < 0.001), whereas for those with Ca intake < or = 800 mg/day, the Fok1 genotype did not have a significant effect on whole body BMD (p = 0.40). The Fok1 genotype was significantly related to the changes during the year in whole body calcium accretion, with the ff genotype having a 63 +/- 20 mg/day deficit compared with the FF genotype (p = 0.008). CONCLUSIONS: The Fok1 polymorphism of the VDR receptor seems to directly affect bone mineral accretion during pubertal growth through an effect on calcium absorption. The relationship between different genetic polymorphisms and bone mineral metabolism may vary by life stage as well as diet. PMID- 15883635 TI - Growth patterns at distal radius and tibial shaft in pubertal girls: a 2-year longitudinal study. AB - Bone changes, in terms of both size and BMD, were assessed longitudinally in pubertal girls. Before puberty, BMD at the distal radius declined, whereas bone size increased, suggesting that normal growing girls experience a transient period of increased bone fragility. This could explain the elevated low-trauma forearm fracture rates reported in earlier studies. INTRODUCTION: Longitudinal data on bone growth during puberty are sparse. Such information is needed to understand the sequence of biological changes, the physical and mechanical consequences for the growing skeleton, and the implications for later life. MATERIALS AND METHODS: The geometric properties and volumetric BMD (vBMD) of the distal radius and tibial shaft were measured using pQCT in 258 pubertal girls followed over 2 years. A new hierarchical linear statistical modeling approach was used to determine true longitudinal trends. RESULTS: The growth rates of cross-sectional area (CSA) and BMC of the distal radius peaked at 16 and 9 months before menarche, respectively. This growth asynchrony between bone size and mass meant that total vBMD of the distal radius declined until 1 year before menarche. At the tibial shaft, total vBMD and cortical vBMD increased monotonically without any such transient reduction. Cortical thickness increased linearly, which was accounted for mainly by bone formation at the periosteal surface before menarche, but by both periosteal and endocortical apposition after menarche. During puberty, the ratio of cortical-to-total CSA of the tibial shaft increased and that of marrow-to-total CSA decreased. CONCLUSIONS: The temporal pattern of bone growth during puberty differs at the distal radius and tibial shaft. A transient decrease in vBMD, arising from asynchronous bone size and mass growth, occurs only at the radius. In the tibia, the mechanism of cortical thickening changes from periosteal apposition premenarche to both periosteal and endocortical apposition postmenarche. PMID- 15883636 TI - Early changes in biochemical markers of bone formation predict BMD response to teriparatide in postmenopausal women with osteoporosis. AB - The relationship between early changes in biochemical markers of bone turnover and the subsequent BMD response to daily teriparatide therapy in women with postmenopausal osteoporosis was studied. Changes in five biochemical markers, obtained from a subset of women enrolled in the Fracture Prevention Trial, were examined. Early increases in the PICP and the PINP were the best predictors of BMD response to teriparatide in this analysis. INTRODUCTION: Early reductions in biochemical markers of bone turnover with antiresorptive therapy negatively correlate with subsequent increases in BMD. We undertook this analysis to determine if early changes in biochemical markers with teriparatide therapy predict subsequent increases in BMD. MATERIALS AND METHODS: In the Fracture Prevention Trial, 1637 postmenopausal women with osteoporosis were randomized to receive daily, self-administered, subcutaneous injections of placebo, teriparatide 20 microg/day, or teriparatide 40 microg/day. Serum concentrations of two bone formation markers (bone-specific alkaline phosphatase [bone ALP] and the carboxy-terminal extension peptide of procollagen type 1 [PICP]) and urinary concentrations of two bone resorption markers (free deoxypyridinoline [DPD] and N terminal telopeptide [NTX]) were assessed in a trial population subset (n = 520) at baseline and at 1, 3, 6, and 12 months. We also assessed serum concentrations of another bone formation marker, the amino-terminal extension peptide of procollagen type 1 (PINP), in a subset of 771 women at baseline and 3 months. Lumbar spine (LS) BMD was measured by DXA at baseline and 18 months. Femoral neck BMD was measured at baseline and 12 months. RESULTS AND CONCLUSION: Baseline bone turnover status correlated positively and significantly with BMD response. The highest correlations occurred for the LS BMD response to teriparatide 20 microg/day. Among all studied biochemical markers, increases in PICP at 1 month and PINP at 3 months correlated best with increases in LS BMD at 18 months (0.65 and 0.61, respectively; p < 0.05). The relationships between these two biochemical markers and the LS BMD response were stronger than the corresponding relationships for the femoral neck BMD response. Using receiver operator curve analysis, we determined that the increases in PICP at 1 month and PINP at 3 months were the most sensitive and accurate predictors of the LS BMD response. PMID- 15883637 TI - Effect of alendronate on the age-specific incidence of symptomatic osteoporotic fractures. AB - Analyses of data from 3658 postmenopausal women with osteoporosis enrolled in the Fracture Intervention Trial showed that alendronate is effective in reducing the risk of symptomatic osteoporotic fractures across a spectrum of ages. INTRODUCTION: Most osteoporosis studies examine the relative risk of fracture based on the entire duration of treatment. Because older patients tend to be at higher risk for osteoporosis-related fractures, this analysis examined the effect of alendronate treatment on the relative risk of fracture in terms of the age that patients attained during the study. MATERIALS AND METHODS: We studied 3658 postmenopausal women with osteoporosis 55-80 years of age at baseline enrolled in the Fracture Intervention Trial, a large randomized, double-blind, placebo controlled study. Patients were treated with placebo or with alendronate at a daily dose of 5 mg for 2 years followed by 10 mg for an additional 1-2.5 years, and monitored for clinical fractures. Age, rather than study time, was the dynamic variable in our analysis. RESULTS: The relative risk reductions for hip, clinical spine, and wrist fractures were constant across age groups, without evidence of a decline at older ages. Specifically, alendronate reduced the risk of clinical fracture by 53% at the hip (relative risk [RR] = 0.47; 95% CI = 0.27 0.81; p < 0.01), 45% at the spine (RR = 0.55; 95% CI = 0.37-0.83; p < 0.01), and 31% at the wrist (RR = 0.69; 95% CI = 0.50-0.98; p = 0.038). In addition, alendronate produced a significant risk reduction of 40% (RR = 0.60; 95% CI = 0.47-0.77; p < 0.01) for the composite event of clinical hip, spine, and wrist fractures. As a consequence of the constant relative risk model, the absolute risk reduction with alendronate treatment increased with age because of the age related increase in fracture risk in the placebo group. The absolute risk reduction for the composite event (hip, spine, and wrist fractures together) for alendronate treatment versus placebo was 65, 80, 111, and 161 women with fractures per 10,000 PYR for the 55 to <65, 65 to <70, 70 to <75, and 75-85 year age groups, respectively. CONCLUSIONS: These data show that alendronate is effective in reducing the risk of symptomatic osteoporotic fractures across a spectrum of ages. The effectiveness is somewhat greater in patients with femoral neck T score < or = -2.5 than in those with a T score < or = -2.0. PMID- 15883638 TI - Controlled trial of pamidronate in children with types III and IV osteogenesis imperfecta confirms vertebral gains but not short-term functional improvement. AB - Bisphosphonates have been widely administered to children with OI based on observational trials. A randomized controlled trial of q3m intravenous pamidronate in children with types III and IV OI yielded positive vertebral changes in DXA and geometry after 1 year of treatment, but no further significant improvement during extended treatment. The treated group did not experience significantly decreased pain or long bone fractures or have increased motor function or muscle strength. INTRODUCTION: Bisphosphonates, antiresorptive drugs for osteoporosis, are widely administered to children with osteogenesis imperfecta (OI). Uncontrolled pamidronate trials in OI reported increased BMD, vertebral coronal area, and mobility, and decreased pain. We conducted a randomized controlled trial of pamidronate in children with types III and IV OI. MATERIALS AND METHODS: This randomized trial included 18 children (4-13 years of age) with types III and IV OI. The first study year was controlled; 9 children received pamidronate (10 mg/m2/day IV for 3 days every 3 months). Four children in each group also received recombinant growth hormone (rGH) injections (0.06 mg/kg/day for 6 days/week). Seven children in the treatment group received pamidronate for an additional 6-21 months. All patients had L1-L4 DXA, spine QCT, spine radiographs, and musculoskeletal and functional testing. RESULTS: In the controlled phase, treated patients experienced a significant increase in L1-L4 DXA z score (p < 0.001) and increased L1-L4 mid-vertebral height (p = 0.014) and total vertebral area (p = 0.003) compared with controls. During extended treatment, DXA z scores and vertebral heights and areas did not increase significantly beyond the 12-month values. Fracture rate decreased significantly in the upper extremities (p = 0.04) but not the lower extremities (p = 0.09) during the first year of treatment. Gross motor function, muscle strength, and pain did not change significantly during the controlled or extended treatment phases. CONCLUSIONS: A controlled trial confirmed the spine benefits of short term pamidronate treatment in children with types III and IV OI. Pamidronate increased L1-L4 vertebral DXA and decreased vertebral compressions and upper extremity fractures. Vertebral measures did not improve during the extended treatment phase. The treatment group did not experience decreased lower extremity long bone fractures, significant improvement in growth, ambulation, muscle strength, or pain. There was substantial variability in individual response to treatment. PMID- 15883639 TI - Growth hormone injections improve bone quality in a mouse model of osteogenesis imperfecta. AB - Systemic growth hormone injections increased spine and femur length in a mouse model of OI. Femur BMC, cross-sectional area, and BMD were increased. Smaller gains were produced in vertebral BMC and cross-sectional area. Biomechanical testing showed improvements to structural and material properties in the femur midshaft, supporting expanded testing of growth hormone therapy in children with OI. INTRODUCTION: Osteoblasts in heterozygous Cola2oim mutant mice produce one half the normal amounts of the alpha2 strand of type I procollagen. The mice experience a mild osteogenesis imperfecta (OI) phenotype, with femurs and vertebrae that require less force than normal to break in a biomechanical test. MATERIALS AND METHODS: Subcutaneous injections of recombinant human growth hormone (rhGH) or saline were given 6 days per week to oim/+ mice between 3 and 12 weeks of age, in a protocol designed to simulate a trial on OI children. RESULTS: rhGH injections promoted significant weight gain and skeletal growth compared with saline-treated control animals. Femur and spine lengths were increased significantly. Significant increases at the femur midshaft in cortical BMD (2.2%), BMC (15.5%), and cross-sectional area (13%) were produced by rhGH treatment. Increases in the same cortical bone parameters were measured in the metaphyseal region of the femur and in tail vertebrae, but lumbar vertebrae showed significant increases in BMC (9.6%) and cross-sectional area (10.1%) of trabecular bone. Three-point bending testing documented functional improvements to the femur mid-shafts. GH treatment produced significant increases in bone stiffness (23.7%), maximum load (30.8%), the energy absorbed by the femurs to the point of maximum load (44.5%), and the energy to actual fracture (40.4%). The ultimate stress endured by the bone material was increased by 14.1%. CONCLUSIONS: Gains in bone length, cross-sectional area, BMD, BMC, structural biomechanical properties, and strength were achieved without directly addressing the genetic collagen defect in the mice. Results support expanded clinical testing of GH injections in children with OI. PMID- 15883640 TI - Leptin treatment induces loss of bone marrow adipocytes and increases bone formation in leptin-deficient ob/ob mice. AB - Normal mice and leptin-deficient ob/ob mice were treated with leptin to study effects on osteogenesis and adipogenesis in bone marrow. Leptin treatment significantly decreased bone marrow adipocyte size and number in ob/ob mice while increasing bone formation, BMC, and BMD. The results suggest that, in leptin sensitive animals, the reduction in marrow adipocytes has positive effects on bone formation. INTRODUCTION: Adipocytes, osteoblasts, and osteoclasts have leptin receptors, and leptin can also affect bone metabolism indirectly through its receptors in the hypothalamus. We examined the effects of leptin treatment on bone formation, BMD, and marrow adipocyte population in normal mice and leptin deficient ob/ob mice. MATERIALS AND METHODS: At the age of 15 weeks, mice were implanted with Alzet osmotic pumps for subcutaneous delivery of treatment solutions (saline, 2.5 microg leptin/day, or 10 microg leptin/day) for 14 days at a delivery rate of 0.25 microl/h. Bone formation was assessed using fluorochrome labels, cell populations were quantified using histomorphometry, and bone densitometry was measured using DXA. We also used a Luminex Beadlyte assay system to quantify cell survival markers in bone marrow samples. RESULTS AND CONCLUSIONS: Results indicate that both doses of leptin decreased the number of marrow adipocytes in ob/ob mice by >20% (p < 0.05) compared with PBS-treated ob/ob mice. The decrease in adipocyte number with leptin treatment is accompanied by an increase in concentration of the apoptosis marker caspase-3 in bone marrow adipocytes and hematopoietic cells. Both leptin doses also significantly (p < 0.05) increased the percentage of fluorochrome-labeled tibial endosteal surface by >30% compared with PBS-treated ob/ob mice. Leptin treatment increased whole body BMC by >30% in the ob/ob mice receiving the highest leptin dose. Leptin treatment provided no increase in bone formation, BMC, or BMD in normal, leptin replete mice. PMID- 15883641 TI - Reduced expression of platelet endothelial cell adhesion molecule-1 in bone marrow cells in mice after skeletal unloading. AB - One week of tail suspension significantly decreased the expression of PECAM-1 in mouse tibial bone marrow cells but not those of a number of other vascular factors. Anti-PECAM-1 antibody suppressed both ALP+ CFU-f formation and ALP production under co-culture of the osteoblastic cell line and the PECAM-1+ endothelial cell line. This study suggests that the reduced ALP activity after skeletal unloading is related to downregulation of PECAM-1 expression in bone marrow cells in mice. INTRODUCTION: Vascular factors play a role in bone development and regeneration. We tested the hypothesis that skeletal unloading reduces osteogenic potential by inhibiting the molecules related to angiogenesis and/or vasculogenesis in bone marrow cells. MATERIALS AND METHODS: Eight-week-old male mice were assigned to three groups after acclimatization for 1 week: ground control (GC), tail suspension (TS), and reloading after 7-day TS (RL). Bilateral tibial and humeral samples were used for analyses. MC3T3-E1, a mouse osteoblastic cell line, and EOMA and ISOS-1, mouse endothelial cell lines, were also used. RESULTS: Flow cytometric analysis revealed that 7-day TS significantly decreased the expression of platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) in tibial bone marrow cells, but not those of angiopoietin-1, angiopoietin-2, Flk 1 (vascular endothelial growth factor receptor-2), and vascular endothelial cadherin. The expression of PECAM-1 in tibial marrow cells was reduced at day 3 of TS to 80% and still showed significantly low levels at day 7 of TS to 72% of that at the respective days of GC. This decreased expression of PECAM-1 after 7 day TS showed the GC level at 5-day reloading after 7-day TS. However, the expression of PECAM-1 in humeral marrow cells (internal bone marrow control) after TS and RL remained unchanged and equivalent to that of GC. The expression level of PECAM-1 mRNA was significantly lower at day 7 of TS to 62% of that in GC. Double labeling analyses revealed that PECAM-1+ cells mostly consisted of endothelial cells and partially of granulocytes. In bone marrow cell cultures, the formation of alkaline phosphatase (ALP)+ colony forming units-fibroblastic was significantly reduced in the presence of anti-PECAM-1 antibody in the medium compared with the presence of immunoglobulin G (0.025 times as much as ALP production with immunoglobulin G). ALP production by cultured MC3T3-E1 was enhanced in combination with PECAM-1+ EOMA (1.8 times as much as ALP production by MC3T3-E1 alone), but not in combination with PECAM-1- ISOS-1. Anti-PECAM-1 antibody inhibited the increase in ALP production under co-culture with EOMA. CONCLUSIONS: Our data show that the reduced ALP activity after skeletal unloading is closely correlated with reduced expression of PECAM-1 in bone marrow cells. We speculate that the loss of osteogenic potential after skeletal unloading is caused by the suppression of PECAM-1 signaling on endothelial cellular surface. PMID- 15883642 TI - Expression patterns of matrix metalloproteinases and vascular endothelial growth factor during epiphyseal ossification. AB - In situ hybridization studies allowed for the localization of three MMPs and the angiogenic factor VEGF during secondary ossification. MMPs were widely expressed during ossification of the secondary center, whereas expression of VEGF was restricted to later stages. INTRODUCTION: The spatiotemporal expression patterns of the matrix metalloproteinases gelatinase-B (MMP-9), collagenase-3 (MMP-13), and membrane-type 1 metalloproteinase (MMP-14) and the angiogenic peptide vascular endothelial growth factor (VEGF) were studied during development of the proximal epiphysis of the rat tibia. MATERIALS AND METHODS: Cell expression was analyzed by in situ hybridization. Studies on osteoclastic activity, matrix mineralization, cell proliferation, and vascular progression were also performed. RESULTS: MMP-9, MMP-13, and MMP-14 were expressed in discrete perichondrial cells that gave way to sites of intrachondral canal formation. High expression levels for the three MMPs were found at the blind ends of advancing intrachondral canals and at the expanding borders of the marrow space. Signals for MMP-9 and MMP-13 were in close proximity but did not overlap, whereas MMP-14 was expressed in both MMP-9+ and MMP-13+ cells. VEGF was not expressed during formation of intrachondral vascular canals but was observed in hypertrophic chondrocytes during formation of the bone marrow cavity. CONCLUSIONS: Expression of MMPs and VEGF are constant events during development of the secondary ossification center. We propose that MMPs are involved in targeting proteolytic activity during epiphyseal development. VEGF is not expressed during early formation of vascular canals, but it may have a role in the formation of the bone marrow cavity. PMID- 15883643 TI - Rac1/Cdc42 and RhoA GTPases antagonistically regulate chondrocyte proliferation, hypertrophy, and apoptosis. AB - The intracellular signaling pathways controlling chondrocyte physiology are largely unknown. Here we show that the small GTPases, Rac1 and Cdc42, accelerate the rate of chondrocyte differentiation and apoptosis, thereby antagonizing the activity of RhoA. These results identify Rac1 and Cdc42 pathways as novel regulators of cartilage development. INTRODUCTION: Proliferation, hypertrophic differentiation, and ultimate apoptosis of chondrocytes regulate endochondral bone growth and development, but the intracellular signaling pathways controlling chondrocyte biology are incompletely understood. In this study, we investigated the role of the small GTPases Rac1 and Cdc42 in chondrocytes. MATERIALS AND METHODS: Rac1 and Cdc42 expression during chondrogenic differentiation was assessed by RT-PCR and Western blotting. Effects of Rac1 and Cdc42 on parameters of chondrocyte biology were studied using transient transfections into primary mouse chondrocytes and stable transfections of the chondrogenic cell line ATDC5. Luciferase assays, RT-PCR, cell proliferation, alkaline phosphatases assays, staining procedures, TUNEL assays, and caspase activity assays were performed to study the chondrocyte response to overexpression of Rac1 and Cdc42 proteins. Activation of the p38 pathway was analyzed using Western blotting with phospho specific antibodies, and mitogen-activated protein (MAP) kinase pathways were inhibited using pharmacological approaches. RESULTS AND CONCLUSIONS: Rac1 and Cdc42 activities are required for maximal activity of the collagen X promoter, a hypertrophic marker, in primary chondrocytes, suggesting essential roles of these GTPases in chondrocyte hypertrophy. Overexpression of Rac1 or Cdc42 in chondrogenic ATDC5 cells results in reductions in cell numbers and marked acceleration of hypertrophic differentiation, thus opposing the effects of the related GTPase RhoA. Rac1 and Cdc42 also induce accelerated chondrocyte apoptosis, as shown by TUNEL and caspase activity assays and changes in cell morphology and actin organization. Rac1 and Cdc42 overexpression results in activation of the p38 MAP kinase pathway in ATDC5 cells, and pharmacological inhibition of p38 signaling blocks the effects of Rac1 and Cdc42 overexpression on hypertrophy and apoptosis. Our results therefore suggest that Rac1 and Cdc42 signaling accelerates progression through the chondrocyte life cycle in a p38 dependent fashion and antagonizes RhoA signaling pathways in chondrocyte proliferation, hypertrophy, and apoptosis. PMID- 15883644 TI - Enamel matrix protein interactions. AB - The recognized structural proteins of the enamel matrix are amelogenin, ameloblastin, and enamelin. While a large volume of data exists showing that amelogenin self-assembles into multimeric units referred to as nanospheres, other reports of enamel matrix protein-protein interactions are scant. We believe that each of these enamel matrix proteins must interact with other organic components of ameloblasts and the enamel matrix. Likely protein partners would include integral membrane proteins and additional secreted proteins. INTRODUCTION: The purpose of this study was to identify and catalog additional proteins that play a significant role in enamel formation. MATERIALS AND METHODS: We used the yeast two-hybrid assay to identify protein partners for amelogenin, ameloblastin, and enamelin. Once identified, RT-PCR was used to assess gene transcription of these newly identified and potential "enamel" proteins in ameloblast-like LS8 cells. RESULTS: In the context of this yeast assay, we identified a number of secreted proteins and integral membrane proteins that interact with amelogenin, ameloblastin, and enamelin. Additionally, proteins whose functions range from the inhibition of soft tissue mineralization, calcium ion transport, and phosphorylation events have been identified as protein partners to these enamel matrix proteins. For each protein identified using this screening strategy, future studies are planned to confirm this physiological relationship to biomineralization in vivo. CONCLUSION: Identifying integral membrane proteins of the secretory surface of ameloblast cells (Tomes' processes) and additional enamel matrix proteins, based on their abilities to interact with the most abundant enamel matrix proteins, will better define the molecular mechanisms of enamel formation at its most rudimentary level. PMID- 15883645 TI - Identification of novel genetic loci for bone size and mechanosensitivity in an ENU mutant exhibiting decreased bone size. AB - Using a dominant ENU mutagenesis screen in C57BL/6J (B6) mice to reveal gene function, we identified a mutant, 917M, with a reduced bone size phenotype, which is expressed only in males. We show that mutation results in osteoblasts with reduced proliferation, increased apoptosis, and an impaired response to in vitro mechanical load. The mutation is mapped to a novel locus (LOD score of 7.9 at 10.5 cM) on chromosome 4. INTRODUCTION: Using a dominant ENU mutagenesis screen in C57BL/6J (B6) mice to reveal gene function, we identified a mutant, 917M, with a reduced bone size phenotype, which is expressed only in males. In this report, we show the chromosomal location of this mutation using linkage analysis and cellular characterization of the mutant phenotype. MATERIALS AND METHODS: The mutant mouse was bred to wildtype B6 to produce progeny for characterization of the bone size phenotype. Periosteal osteoblasts isolated from the tibia and femur of mutant and wildtype mice were studied for proliferation, differentiation, and apoptosis potential. To determine the chromosomal location of the mutation, a low resolution linkage map was established by completing a genome-wide scan in B6C3H F2 male mice generated from intercross breeding of mutant mice. RESULTS AND CONCLUSIONS: Mutant progeny (16 weeks old) displayed a total body bone area that was 10-13% lower and a periosteal circumference that was 5-8% lower at the femur and tibia midshaft compared with wildtype B6 mice. Periosteal osteoblasts from mutant mice showed 17-27% reduced cell proliferation and 23% increased apoptosis compared with wildtype controls. In addition, osteoblasts from mutant mice showed an impaired response to shear stress-induced proliferation rate, an in vitro model for mechanical loading. Interval mapping in B6C3H F2 males (n = 69) indicated two major loci affecting bone size on chromosome 1 at 45 cM (LOD 4.9) and chromosome 4 at 10.5 cM (LOD 7.9, genome-wide p < 0.01). Interval mapping using body weight as covariate revealed only one significant interval at chromosome 4 (LOD 6.8). Alleles of the chromosome 4 interval inherited from the B6 mutant strain contributed to a significantly lower bone size than those inherited from C3H. A pairwise interaction analysis showed evidence for a significant interaction between loci on chromosome 1 with the chromosome 4 quantitative trait loci. The 917M locus on chromosome 4 seems to be novel because it does not correspond with those loci previously associated with bone size on chromosome 4 in B6 and C3H/HeJ mice or other crosses. PMID- 15883646 TI - PTHrP signaling targets cyclin D1 and induces osteoblastic cell growth arrest. AB - PTHrP control of the MC3T3-E1 cell cycle machinery showed that, during differentiation, PTHrP induced G1 growth arrest. Cyclin D1 was a critical mediator as a downstream effector of cAMP, PKC, and MAPK signaling, and the process was PKA-independent. The involvement of JunB has been found critical for PTHrP effects. INTRODUCTION: PTH-related protein (PTHrP) has been implicated in the control of bone cell turnover, but the mechanisms underlying its effect on osteoblast proliferation and differentiation have not been clearly defined. The mechanisms by which PTHrP impacts cell cycle proteins and the role of signaling pathways in differentiated osteoblasts were studied. MATERIALS AND METHODS: To elucidate the role of PTHrP, flow cytometric analyses were performed using MC3T3 E1 and primary mouse calvarial cells. Relative protein abundance (Western blot), physical association of partners (immunoprecipitation), and kinase activities (in vitro kinase assays using either GST-Rb or H1-histone as substrates) of cell cycle-associated proteins in vehicle and PTHrP-treated 7-day differentiated cells were determined. ELISA and/or Northern blot analyses were done to evaluate JunB and cyclin D1 expression. SiRNA-mediated gene silencing experiments were performed to silence JunB protein. Finally, inhibitors of cAMP, protein kinase A (PKA), protein kinase C (PKC), and mitogen-activated protein kinase (MAPK) were used to determine involvement of different signaling pathways. RESULTS: PTHrP inhibited cyclin D1 protein expression 7-fold in a dose- and time-dependent manner and increased the level of p16 protein in differentiated osteoblasts. Additionally, PTHrP reduced cyclin D1-CDK4/CDK6 and CDK1 kinase activities. Forskolin, a cAMP agonist, mimicked PTHrP action, and the PKC inhibitor, GF109203X, slightly blocked downregulation of cyclin D1, implying involvement of both cAMP and PKC. U0126, a MAPK inhibitor, alone decreased cyclin D1 protein, suggesting that the basal cyclin D1 protein is MAPK dependent. H-89, a PKA inhibitor, did not alter the effect of PTHrP on cyclin D1, suggesting a PKA independent mechanism. Finally, expression of JunB, an activating protein-1 transcription factor, was significantly upregulated, and silencing JunB (siRNA) partially reversed the cyclin D1 response, implying involvement of JunB in the PTHrP-mediated growth arrest of MC3T3-E1 cells. CONCLUSION: PTHrP upregulates JunB and reduces cyclin D1 expression while inducing G1 cell cycle arrest in differentiated osteoblasts. Such regulation could be an important determinant of the life span and bone-forming activity of osteoblasts. PMID- 15883648 TI - Affiliative relations among male Japanese macaques (Macaca fuscata yakui) within and outside a troop on Yakushima Island. AB - Male Japanese macaques (Macaca fuscata yakui) in a troop on Yakushima Island frequently groom other males. However, previous studies have not compared the social relations of troop males to those of non-troop males. I followed all troop males and non-troop males in and near a troop during a mating season and during the following non-mating season and recorded their neighbors, grooming, and agonistic interactions. Comparisons of the social relations of troop males and non-troop males with other troop members revealed that grooming and agonistic interactions with females during the mating season were similar between troop and non-troop males. However, troop males groomed each other more often and had fewer agonistic interactions among themselves than did non-troop males. Compared to what occurred in the mating season, troop males groomed females less often and exchanged grooming bouts more often with other troop males during the non-mating season. One non-troop male groomed females more frequently than did any troop male in both seasons, and this male groomed troop males more frequently than did any troop male in the non-mating season. This male immigrated into the troop during the following mating season. Regardless of their competition with respect to reproduction, male Japanese macaques on Yakushima Island maintain affiliative relations, probably to cooperatively defend fertile females from non-troop males. PMID- 15883649 TI - The formation of the brush-sticks: modification of chimpanzees or the by-product of folding? AB - Based on field research and experimental treatments of trees, we investigated the formation of the brush-like shape of digging sticks used by chimpanzees (Pan troglodytes troglodytes). Evidence obtained in the field consisted of digging sticks found in Mboete, Equatorial Guinea, which is a newly reported locality for this type of tool, and Campo, Cameroon. Digging sticks used by chimpanzees in these areas had a brush-like shape at one end, which was quite different from the other end that was probably used for digging. In our tree-breaking experiment, 8 out of 17 species acquired a typical brush-like shape without human modification when broken off, and the shapes of the stumps were similar to those found in the field. Other species did not acquire the brush-like shape naturally or even after human modifications, and the stumps had different shapes from those found in the field. Our findings suggest that the brush-like shapes of digging sticks are often naturally formed when broken off from trees, depending on the nature of the fibre structure, and that the brush-like end is not used as the digging tool. We conclude that the vegetation surrounding termite mounds might influence how chimpanzees combine different types of tools, i.e., digging stick, brush-stick and fishing tool, for obtaining termites. PMID- 15883650 TI - Building a residential treatment program for dually diagnosed women with their children. AB - The epidemic of drug and alcohol abuse in our nation impacts millions of women, mothers, and children. Addicted mothers with complex problems and numerous co morbidities present unique treatment challenges. This intergenerational cycle of abuse and addiction is difficult to stop. Arkansas CARES (Center for Addictions Research Education and Services, referred to in this article as CARES) initially was created to treat addicted pregnant and postpartum women and their infants. CARES evolved into a residential treatment program for dually diagnosed mothers with their children. This paper is a synopsis of a presentation delivered at the North American Society for Psychosocial Obstetrics and Gynecology. It shares a glimpse inside the treatment program and lessons learned along the way in an effort to assist others who are interested in building treatment programs for addicted women with their children. PMID- 15883651 TI - Prevalence of suicidality during pregnancy and the postpartum. AB - This review examined the available prevalence estimates of suicidality (suicide deaths, attempts, and ideation including thoughts of self harm) in pregnancy and the postpartum. Studies that used defined community or clinic samples were identified through multiple electronic databases and contacts with primary authors. Definitions of and measurement of suicide deaths, intentional self harming behavior, suicide attempts, and thoughts of death and self-harm were varied and are described with each study. While suicide deaths and attempts are lower during pregnancy and the postpartum than in the general population of women, when deaths do occur, suicides account for up to 20% of postpartum deaths. Self-harm ideation is more common than attempts or deaths, with thoughts of self harm during pregnancy and the postpartum ranging from 5 to 14%. The risk for suicidality is significantly elevated among depressed women during the perinatal period, and suicide has been found to be the second or leading cause of death in this depressed population. PMID- 15883652 TI - Depressive symptoms in early pregnancy, two months and one year postpartum prevalence and psychosocial risk factors in a national Swedish sample. AB - BACKGROUND: Depression and other psychiatric disorders during pregnancy and postpartum is an important health problem, especially if the symptoms are recurrent or sustained. METHODS: All Swedish speaking women attending their first antenatal care visit during three predestined weeks were invited to participate. Depressive symptoms were evaluated using the Edinburgh Postnatal Depression Scale (EPDS) in early pregnancy, two months and one year postpartum. RESULTS: In all, 2430 women completed three questionnaires. A dose-effect relation was found between the numbers of stressful life events experienced in the year prior to pregnancy and mean EPDS score in pregnancy. The prevalence of recurrent or sustained depressive symptoms (EPDS> or =12 on all three evaluations) was 3% (79/2430). Three factors were associated with depressive symptoms, two or more stressful life events in the year prior to pregnancy, native language other than Swedish and unemployment. CONCLUSIONS: Apart from questions about psychiatric history, a psychosocial history in early pregnancy including stressful life events, native language and employment status could help the health professionals to identify women at risk for recurrent or sustained depression during pregnancy and the year after giving birth. PMID- 15883653 TI - Validation of the Edinburgh postnatal depression scale (EPDS) in a sample of women with high-risk pregnancies in France. AB - This research is intended to validate the Edinburgh Postnatal Depression Scale (EPDS) in a high-risk pregnant population. METHOD: Sixty women attending antenatal consultations for pregnancy complication in a major Parisian maternity facility were included. They completed the EPDS and were then interviewed according to a standardised psychiatric interview. RESULTS: The study of its sensitivity, specificity and predictive values, with a DSM-IV diagnosis of major depression as the reference, found that 11.5 was the optimal cut-off score (Se 0.80; Sp 0.80). Its validity as an index of severity of depression was also good as well as internal consistency and reliability. Factor analysis showed that its internal structure is composed of two subscales (F2 "depression" and F1 with items reflecting depression and other disorders, including anxiety). CONCLUSION: The French version of the EPDS would be a valid instrument to identify pregnant women who are likely to have clinical major depression. The results may have to be confirmed on a community sample before clinical use. PMID- 15883654 TI - The protein ICP0 of herpes simplex virus type 1 is targeted to nucleoli of infected cells. Brief report. AB - This study describes the nucleolar localization of the viral protein ICP0 of herpes simplex virus type 1. We show that the RING finger domain of ICP0 is essential for ICP0 to localize in nucleoli of transfected and 4 hour-infected cells. ICP0 forms particular intranucleolar domains that do not correspond to any known nucleolar domains. This distribution was confirmed by immunoblots performed on fractionated infected cells. Quantitative RT-PCR experiments indicated that ICP0 did not increase the transcription from the RNA polymerase I (Pol I) promoter in transfected cells, an effect opposite to that observed on viral and cellular Pol II promoters. Nucleoli are thus, after PML bodies and centromeres, a novel nuclear structure targeted by ICP0. PMID- 15883655 TI - Molecular characterization of isolates of anagyris vein yellowing virus, plantago mottle virus and scrophularia mottle virus -- comparison of various approaches for tymovirus classification. AB - The complete nucleotide sequences were determined for the genomic RNAs of three tymoviruses, i.e. isolates of anagyris vein yellowing virus (AVYV), plantago mottle virus (PlMoV) and scrophularia mottle virus (SrMV) which are all serologically closely related to ononis yellow mosaic virus (ibid) and to Nemesia ring necrosis virus (NeRNV), a recently described recombinant virus which is widely spread in commercially grown ornamental plant species belonging to the Scrophulariaceae. Total nucleotide and coat protein amino acid sequence identities revealed similar groupings in the genus tymovirus as serological studies did. The latter, however, tended to suggest much closer relationships than the molecular data and may fail to recognise the distinctiveness of new tymovirus species. The usefulness of various species demarcation criteria for the classification of tymoviruses is discussed. PMID- 15883656 TI - Identification of a Bohle iridovirus thymidine kinase gene and demonstration of activity using vaccinia virus. AB - In recent years interest in the family Iridoviridae has been renewed by the identification of a number of viruses, particularly from the genus Ranavirus, associated with disease in a range of poikilotherms. Ranaviruses have been isolated from amphibian, piscine and reptilian species. Here we describe an open reading frame (ORF) identified in the genome of Bohle iridovirus (BIV) which contains a nucleotide binding motif conserved within the thymidine kinase (TK) genes of iridoviruses from other genera (lymphocystis disease virus, LCDV, type species of the genus Lymphocystivirus; Chilo iridescent virus, CIV, type species of the genus Iridovirus). The ability of this putative gene to express a functional TK was confirmed by rescue of a TK negative mutant vaccinia virus in the presence of selective media, when expression was controlled by a vaccinia virus promoter. The sequence of the BIV TK was compared with the homologous sequences from epizootic haematopoietic necrosis virus (EHNV), a virus associated with disease in fish, from Wamena iridovirus (WIV) associated with systemic disease in green pythons, and from frog virus 3 (FV3) the ranavirus type species. Comparisons between these sequences and those available from other ranaviruses, other iridoviruses, other DNA viruses and cellular TKs are presented. PMID- 15883657 TI - Multiple gene segment reassortment between Eurasian and American lineages of influenza A virus (H6N2) in Guillemot (Uria aalge). AB - Guillemots banded in the northern Baltic Sea were screened for influenza A virus (IAV). Three out of 26 sampled birds tested positive by RT-PCR. Two of these were characterized as subtype H6N2. Phylogenetic analyses showed that five gene segments belonged to the American avian lineage of IAVs, whereas three gene segments belonged to the Eurasian lineage. Our findings indicate that avian IAVs may have a taxonomically wider reservoir spectrum than previously known and we present the first report of a chimeric avian IAV with genes of American and Eurasian origin in Europe. PMID- 15883658 TI - The complete genome sequence, organization and affinities of carrot red leaf virus. AB - A sequence of 5723 nucleotides (GenBank accession number: AY695933) is reported for the RNA genome of an isolate of Carrot red leaf virus (CtRLV). The sequence is predicted to contain six large open reading frames and non coding sequences of 28 nucleotides at the 5' end, 110 nucleotides at the 3' end, and 215 nucleotides between the two main blocks of coding sequences. The 5' coding region encodes two polypeptides with calculated molecular masses (Mr) of 28.6 kDa (P0) and 68.2 kDa (P1) that overlap in different reading frames. Circumstantially, the third ORF in the 5' block is putatively translated by frameshift read-through to yield a polypeptide (P1 + P2) with a calculated Mr of 116.9 kDa. Frameshifting is predicted at a "shifty" sequence (GGGAAAC; nt 1523-1529) also found in most members of the genus Polerovirus. The C-terminal region of the 116.9 kDa polypeptide includes the consensus sequence for the viral RNA-directed RNA polymerase. The 3' block of coding sequence defines three putative polypeptides of: 23.0 kDa (P3), 21.3 kDa (P4, in a different reading frame) and 77.2 kDa (P3 + P5, by read-through of P3) respectively. From the genome structure of CtRLV, it is suggested that this virus belongs to the genus Polerovirus, rather than either the genus Luteovirus or the genus Enamovirus. PMID- 15883659 TI - Socioeconomic influences on bone health in postmenopausal women: findings from NHANES III, 1988-1994. AB - Our objectives were (1) to examine the associations of education and income with bone health in non-Hispanic White, non-Hispanic Black and Mexican-American postmenopausal women, (2) to determine if any observed associations can be explained by behavioral factors such as calcium intake and physical activity and (3) to determine if government food assistance and education are associated with increased calcium intake among low-income women. Cross-sectional data were gathered by the Third National Health and Nutrition Examination Survey, 1988-1994 (NHANES III) using a stratified multistage probability design. Bone health was indicated by total hip bone mineral density (BMD, g/cm2). Multiple linear regression was used to evaluate the associations of education, income and behavioral factors with BMD. There were 2,905 postmenopausal women with acceptable DXA scans and complete relevant data selected from a nationally representative sample of the civilian non-institutionalized population aged 2 months and older. Education and income were positively associated with BMD in Black and White women, respectively, but not in Mexican-American women. When behavioral factors were included in the analyses, associations with education and income were eliminated. Instead, positive associations with estrogen use, calcium intake and physical activity, and a negative association with smoking, were noted in White women. Among low-income women, education was associated with increased calcium intake, while participation in the Food Stamp Program was associated with increased calcium intake in Black women. We conclude that education and/or income are positively associated with BMD among Black and White postmenopausal women, and that efforts to promote bone health among low-income women are warranted. PMID- 15883660 TI - Hearing sensitivity and bone mineral density in older adults: the Health, Aging and Body Composition Study. AB - Bone mineral density (BMD) may be associated with hearing loss in older adults. Demineralization of the cochlear capsule has been associated with hearing loss in those with Paget's disease of the bone and otosclerosis. Osteoporosis may also result in cochlear capsule demineralization. We hypothesized that lower hip BMD and lower heel ultrasound measurements would be associated with hearing loss in a population-based sample of 2,089 older black and white men and women. Bone parameters and hearing function were measured at the fourth clinical follow-up visit. Audiometric threshold testing was used to measure air- and bone-conduction hearing sensitivity. BMD of the hip and its subregions was measured using dual energy X-ray absorptiometry. Calcaneal bone measurements [broadband ultrasound attenuation (BUA), speed of sound (SOS) and the quantitative ultrasound index (QUI)] were obtained using heel ultrasound. After adjusting for known hearing loss risk factors, no association was found between hearing and any of the bone measurements in whites and black women. In black men, however, lower hip BMD was associated with higher odds of hearing loss; for each standard deviation decrease in total hip BMD, the odds of hearing loss were 1.41 (95% confidence interval 1.08, 1.83), 1.39 (95% CI 1.07, 1.82) for femoral neck BMD and 1.65 (95% CI 1.26, 2.16) for trochanter BMD. Conductive hearing loss was associated with lower heel ultrasound measurements, though only among white men. The results of this study are mixed and inconclusive. Lower BMD of the hip and its subregions was associated with hearing loss among black men, but not among whites or black women. Lower measurements on heel ultrasound were associated with conductive hearing loss, though only among white men. These results suggest that axial and appendicular bone parameters may be modestly associated with hearing loss in older men, but not in women. PMID- 15883661 TI - Comparison of questionnaire and quantitative ultrasound techniques as screening tools for DXA. AB - The aim of the study is to assess the sensitivity and specificity of different techniques and their ability to act as screening tools in relation to dual energy X-ray absorptiometry (DXA) in a group of 208 postmenopausal women. In this study we examined eight screening systems for the diagnosis of osteoporosis, the osteoporosis self-assessment tool (OST), the osteoporosis risk assessment instrument (ORAI), the osteoporosis index of risk (OSIRIS), a risk index derived using data from the study of osteoporotic fractures (SOFSURF), the simple calculated osteoporosis risk estimation (SCORE), patient body weight (pBW), along with two ultrasound based systems, the Sunlight Omnisense (Sunlight Medical, Rehovot, Israel) and the CUBA Clinical (McCue plc, Winchester, UK). The sensitivity and specificity of the different techniques in relation to DXA were plotted as receiver-operating characteristic (ROC) curves at three different levels (DXA T-score -2.5 osteoporosis, -2 and -1 osteopenia). The areas under the curves (AUC) were calculated and showed broadband ultrasound attenuation (BUA) at the calcaneus to provide consistently the highest AUC (0.77-0.81). The velocity of sound (VOS) of the calcaneus (AUC=0.72-0.76) was equally good, but was out performed by some of the questionnaire systems (AUC=0.66-0.79). Both the questionnaire systems and the CUBA Clinical out-perform the Sunlight Omnisense (AUC=0.58-0.7), which showed comparable performance with body weight (AUC=0.66 0.69). The results show that QUS is capable of selecting patients with low bone density as measured by DXA. A patient displaying a low QUS value should be followed up with a DXA scan to confirm the diagnosis. PMID- 15883662 TI - The effect of intravenous pamidronate versus oral alendronate on bone mineral density in patients with osteoporosis. AB - Intravenous pamidronate is frequently used for the treatment of osteoporosis in patients who cannot tolerate oral bisphosphonates. The aim of the present study was to compare the changes in bone mineral density (BMD) after 1 year of treatment with either oral alendronate or intravenous pamidronate in patients with osteoporosis. We studied 40 consecutive patients starting treatment for osteoporosis: 20 received oral alendronate 10 mg/day and 20 received intravenous pamidronate 60 mg/3 months. Patients were started on intravenous pamidronate in the case of intolerance (within 1 month of start of treatment) of an oral bisphosphonate or in the case of contraindications for an oral bisphosphonate. BMD (spine and total hip) was measured with dual X-ray absorptiometry (DEXA) at the start of treatment and after 1 year. The BMD of the lumbar spine increased by 4.0% (P<0.05 vs baseline) in both groups, and the BMD of the hip increased by 3.3% and 2.9% (P<0.05 vs baseline) in the alendronate and pamidronate groups, respectively. The increases in BMD of the vertebral spine and the total hip after 1 year are comparable in the alendronate and pamidronate groups. We conclude that intravenous pamidronate can be used successfully as an alternative treatment in patients with gastrointestinal intolerance of an oral bisphosphonate. PMID- 15883663 TI - The effects of thermal annealing on the relaxation of rubbing-induced birefringence in polystyrene. AB - We have conducted a systematic study on the effects of post rubbing annealing on the relaxation of rubbing-induced birefringence of polystyrene. It is found that annealing at T(0) only affects the relaxation up to T(0)+T(Lag), where T(Lag) is proportional to the logarithm of the annealing time t(A). A theoretical model based on the distribution of relaxation times due to the individual birefringence elements is proposed. To remove its contribution to the net birefringence each element must overcome an energy barrier E=(317+1.17xi)x10(3) J/mol, and therefore must have a characteristic relaxation time tau which depends on temperature T and a barrier height which ranges from 340.4 kJ/mol to 445.7 kJ/mol. The relaxation of birefringence is expressed by the equation NB(T, t)=[Formula: see text]N(xi)e( t/tau(T,xi))dxi, in which both the relaxation time tau(T,xi) and the distribution function N(xi) can be extracted from experimental data. The predictions of the model agree well with all the experimental results presented in this work. The differences and similarities of the relaxation of birefringence with respect to the physical aging of quenched PS are discussed. In particular, similarities in terms of the general temperature lag phenomena are noted. PMID- 15883664 TI - Use of ligasure in thyroidectomy procedures: results of a prospective comparative study. AB - The thyroidectomy procedure requires many manupulations to achieve prompt hemostasis. This study assessed whether the outcomes of thyroidectomy using the Ligasure electrothermal vessel sealer were comparable with the conventional suture-ligation technique. We prospectively evaluated 58 consecutive patients who underwent thyroidectomy. Patients were allocated into two groups according to their preference. There were 30 patients in the Ligasure group and 28 patients in the conventional surgery group. Complications, operating time, and hospital stay were compared between the two groups and suture-ligations performed in the Ligasure group were recorded. The age, sex, and indications were similar in the two groups (p > 0.05). Complication rates and hospital stays did not show any difference according to the techniques used. Operating time was shorter in the hemithyroidectomy and total thyroidectomy patients of the Ligasure group (mean +/ SD: 77.38 +/- 13.71 vs. 99.80 +/- 12.53 minutes, p = 0.005; and 102.50 +/- 16.69 vs.128.89 +/- 19.74 minutes, p = 0.010). The mean +/- SD number of suture ligations for each patient in the Ligasure group was 1.83 +/- 2.12. Thyroid surgery using the Ligasure is safe, and its complication rates are comparable to these found with the conventional surgical technique. Use of the Ligasure during hemithyroidectomy and total tyhroidectomy operations provides a significantly shorter operating time. PMID- 15883665 TI - Active antigen-specific immunotherapy of melanoma: from basic science to clinical investigation. AB - Advanced-stage melanoma here dismal prognosis, and novel therapeutic approaches are urgently required. The possibility of taking advantage of the immune response of patients for its treatment has been an appealing concept for almost a century. Only during the last decade, however, has the molecular identification of tumor associated antigens (TAAs) offered the possibility of vaccinating patients (e.g., active induction of TAA-specific immune responses). Active antigen-specific immunotherapy (AASIT) is currently being investigated in a number of clinical centers as a treatment option for advanced-stage melanoma. A large number of melanoma TAAs have been molecularly characterized and are being used in vaccination trials in various molecular forms and according to various immunization protocols. Here we provide a short overview on melanoma TAAs, the technologies currently in use to induce specific cytotoxic T-lymphocyte (CTL) responses in vivo, and their monitoring. We also propose a tentative AASIT agenda for the next few years, aiming at improving the capacity to induce and monitor TAA-specific immune responses and to verify their clinical effectiveness. PMID- 15883666 TI - Unexpected results using rapid intraoperative parathyroid hormone monitoring during parathyroidectomy for primary hyperparathyroidism. AB - Rapid intraoperative parathyroid hormone (RIOPTH) monitoring predicts complete removal of all hypersecreting tissue by means of a significant parathyroid hormone (PTH) decrease. In this study we have tried to provide an explanation for some unexpected results of RIOPTH monitoring observed during a series of 125 conventional parathyroidectomies for primary hyperthyroidism, discussing the possible consequences on the surgical strategy. Three main groups can be recognized: (1) spikes: a PTH increase 10 minutes after removal of the diseased gland was observed in three patients; (2) false-negative results: six patients showed an inadequate PTH decreases at 10 minutes, three of them resulting in cure at 20 minutes (all six patients were cured at follow-up); (3) false-positive results: five patients with multiglandular disease showed a PTH decrease to a cure level despite excision of one adenoma only (in two of these patients a 20 minute sample showed a PTH increase soon after manipulation of the second adenoma). We concluded that the spike, almost certainly a consequence of manipulating the adenoma, when detected should be considered the "true" baseline value. False-negative results are to some extent related to undetected spikes. The assay used for RIOPTH determination and PTH half-life variability may also play a role. A false-negative result usually prolongs the surgical time. False positive results are usually related to a double adenoma, one functionally prevailing over the other. Because in our experience manipulation of the second adenoma brought a PTH increase detected with RIOPTH monitoring, we believe that the second adenoma should be excised. PMID- 15883667 TI - Effects of dietary selenomethionine on larval rainbow trout (Oncorhynchus mykiss). AB - Increased selenium (Se) concentrations in water (>10 microg/L) have been measured in the San Diego Creek, which is a tributary of the Upper Newport Bay in Orange County, CA. The objective of this study was to develop tissue- and dietary-based thresholds for Se in resident fish species in San Diego Creek. A 90-day dietary experiment was conducted to determine the effects of seleno-L-methionine (SeMe) on the growth, survival, and whole-body Se accumulation in larval (24-day-old) rainbow trout. Decreased and oxidized glutathione (GSH-to-GSSG ratio) and thiobarbituric acid-reactive substances (TBARS) were also measured in livers of exposed animals to assess oxidative damage caused by Se. Fish food was spiked with SeMe to contain 4.6, 12, and 18 microg/g (dry weight) of Se. Fish exposed to SeMe for 90 days exhibited a significant decrease in body weight and fork length in the 4.6 and 12 microg/g Se treatments compared with controls. Whole-body total Se concentrations increased significantly in fish fed 12 and 18 microg/g SeMe after 90 days compared with controls. Lipid peroxidation (TBARS) and GSH-to-GSSG ratios were unchanged by SeMe treatment. Based on decreased growth after 90 days, a dietary Se lowest observed-effect concentration (LOEC) value of 4.6 microg/g and a Se body burden LOEC of 1.20 microg/g (wet weight) were estimated. PMID- 15883668 TI - Perfluorinated compounds in aquatic organisms at various trophic levels in a Great Lakes food chain. AB - Trophic transfer of perfluorooctanesulfonate (PFOS) and other related perfluorinated compounds was examined in a Great Lakes benthic foodweb including water-algae-zebra mussel-round goby-smallmouth bass. In addition, perfluorinated compounds were measured in livers and eggs of Chinook salmon and lake whitefish, in muscle tissue of carp, and in eggs of brown trout collected from Michigan. Similarly, green frog livers, snapping turtle plasma, mink livers, and bald eagle tissues were analyzed to determine concentrations in higher trophic-level organisms in the food chain. PFOS was the most widely detected compound in benthic organisms at various trophic levels. Concentrations of PFOS in benthic invertebrates such as amphipods and zebra mussels were approximately 1000-fold greater than those in surrounding water, which suggested a bioconcentration factor (BCF; concentration in biota/concentration in water) of 1000 in benthic invertebrates. Concentrations of PFOS in round gobies were two- to fourfold greater than those in their prey organisms such as zebra mussels and amphipods. Concentrations of PFOS in predatory fishes (Chinook salmon and lake whitefish) were 10 to 20-fold greater than those in their prey species. Concentrations of PFOS in mink and bald eagles were, on average, 5- to 10-fold greater than those in Chinook salmon, carp, or snapping turtles. Because of the accumulation of PFOS in liver and blood, the biomagnification factor (BMF) of perfluorinated compounds in higher trophic-level organisms such as salmonid fishes, mink, and eagles were based on the concentrations in livers or plasma. Overall, these results suggest a BCF of PFOS of approximately 1000 (whole-body based) in benthic invertebrates, and a BMF of 10 to 20 in mink or bald eagles, relative to their prey items. Eggs of fish contained notable concentrations of PFOS, suggesting oviparous transfer of this compound. PFOA was found in water, but its biomagnification potential was lower than that of PFOS. PMID- 15883669 TI - Toxicological guidelines for monocyclic nitro-, amino- and aminonitroaromatics, nitramines, and nitrate esters in drinking water. AB - In order to secure a safe drinking water supply, the setting of tolerable/acceptable ceilings of drinking water hygiene is required with regard to xenobiotics resulting from several anthropogenic impacts. This is done in practice by using drinking water guidelines or standards as quantitative objectives. The list of the new EU Directive or the German drinking-water standards is limited to those parameters that have the highest relevance for drinking water quality; nitro compounds (NCs) are not regulated. Because other substances contained in water can also represent a hazard for human health, the German Drinking Water Ordinance clarifies that specific actions must be implemented if compounds other than those regulated appear at concentrations that may be a cause for concern regarding human health. NCs serve as intermediates for dyes, pharmaceuticals, and synthetic materials; they themselves are used as solvents, explosives, and pesticides. During their commercial production or from their use, they may be released to the environment and lead to a contamination of aquatic systems and thus also of drinking water resources. In practice, therefore, a need for assessment is frequently given for relevant NCs. For 19 nitro-, amino-, and aminonitroaromatics, nitramines, and nitrate esters health based drinking water guide values have been derived. For toxicological evaluation and derivation of guideline values for the NCs of interest, the tolerable daily intake approach was used for chemicals exhibiting a threshold for toxic effects. This was done by using established tolerable body doses for humans based on an identified no-observed-adverse-effect level/low-observed-effect-level for the most sensitive indicator for toxicity. In the case of nonthreshold chemical substances, suitable estimates of excess lifetime cancer risk have been applied. PMID- 15883670 TI - Heavy metal levels in fish from coastal waters of Uruguay. AB - Copper, mercury, and zinc levels were determined in muscle and liver (N = 163) of seven fish species caught in coastal waters off Montevideo and Piriapolis (control site): Odontesthes spp., Mugil platanus, Micropogonias furnieri, Urophycis brasiliensis, Cynoscion guatucupa, Menticirrhus americanus, and Mustelus schmitti. The local population commonly uses these species for consumption. Heavy metal concentrations determined in this study were generally below those obtained for fish caught in Argentinean and Brazilian coastal waters, with some exceptions in the case of mercury and zinc. Based on copper, mercury, and zinc levels in muscle tissue, we conclude that the fish studied here are acceptable for human consumption. Nevertheless, it is recommended not to consume the fish liver (up to 466 microg Zn g(-1) dry weight in liver) nor large specimens of the investigated species. Regional programs involving the neighboring countries should be established to assess the fisheries resources and potential risks for human health. PMID- 15883671 TI - Organochlorine concentrations in bonnethead sharks (Sphyrna tiburo) from Four Florida Estuaries. AB - Because of their persistence in aquatic environments and ability to impair reproduction and other critical physiological processes, organochlorine (OC) contaminants pose significant health risks to marine organisms. Despite such concerns, few studies have investigated levels of OC exposure in sharks, which are fish particularly threatened by anthropogenic pollution because of their tendency to bioaccumulate and biomagnify environmental contaminants. The present study examined concentrations of 29 OC pesticides and total polychlorinated biphenyls (PCBs) in the bonnethead shark (Sphyrna tiburo), an abundant species for which evidence of reproductive impairment has been observed in certain Florida populations. Quantifiable levels of PCBs and 22 OC pesticides were detected via gas chromatography and mass spectrometry in liver of 95 S. tiburo from four estuaries on Florida's Gulf coast: Apalachicola Bay, Tampa Bay, Florida Bay, and Charlotte Harbor. In general, OC concentrations were significantly higher in Apalachicola Bay, Tampa Bay, and Charlotte Harbor S. tiburo in relation to the Florida Bay population. Because the rate of infertility has been shown to be dramatically higher in Tampa Bay versus Florida Bay S. tiburo, the present findings allude to a possible relationship between OC exposure and reproductive health that requires further investigation. Pesticide and PCB concentrations did not appear to significantly increase with growth or age in S. tiburo, suggesting limited potential for OC bioaccumulation in this species compared with other sharks for which contaminant data are available. Concentrations of OCs in serum and muscle were not correlated with those in liver, indicating that these tissues are poor surrogates for measuring internal OC burden in this species via nonlethal sampling procedures. PMID- 15883672 TI - Nonylphenol and nonylphenol ethoxylates in river water, drinking water,and fish tissues in the area of Chongqing, China. AB - Little attention has been paid to the estrogenic-like compounds, such as 4 nonylphenol (4-NP) and its potential precursor nonylphenol ethoxylates (NPEOs), in China although its usage is huge. Water samples and corresponding drinking water samples were seasonally collected at five sites of each of the two main rivers in Chongqing Area. Individual nonylphenol ethoxylates (NPEOs) and 4-NP in the Changjiang River and Jialingjiang River were detected by normal-phase liquid chromatography electrospray ionization mass spectrometry and gas chromatography mass spectrometry. The results indicated that of the five sampling points in the two rivers, NPEOs were the dominant pollutant in April and December with the similar distribution profile, and total NPEOs with different ethylene oxide lengths were 6.9-97.6 microg/L in April and 2.5-52.7 microg/L in December. However, NP was the dominant pollutant in July with a concentration of 1.7-7.3 microg/L. Corresponding drinking water samples derived from river water as source suggested that the conventional water treatment process used in the five waterworks could remove NPEOs from the source water with high removal efficiency (>99%). The 4-NP removal efficiency, however, varied in a range of 62% to 95%, leaving a significantly high concentration of NP (0.1 to 2.7 microg/L) in drinking water in July. Fish samples taken in December 2000 contained 4-NP of approximately 1.9 microg/g and NPEOs of 0.4-48.3 microg/g, with the highest concentration level found in liver. PMID- 15883673 TI - Bisphenol A in ambient air particulates responsible for the proliferation of MCF 7 human breast cancer cells and Its concentration changes over 6 months. AB - To survey the estrogenic activity of the organic extracts from particulate matter of urban ambient outdoor air, samples were collected on glass fiber filters using a high-volume air sampler on the rooftop of our institute for 6 months (six filters/month). After extracting the organic materials and separating them into three fractions, i.e., acidic, neutral, and basic, we applied a cell-growth assay using MCF-7 human breast cancer cells to the original extract and the extracts of the fractions. Only the extract in the acidic fraction showed cell proliferation activity in a dose-response manner. To survey the chemical(s) responsible for the activity, a gas chromatography/mass spectrometry (GC/MS) analysis was conducted after silylating the extract. The presence of bisphenol A (BPA) was confirmed, because the retention times and the MS fragment patterns between the silylated derivative of a component in the sample and that of BPA itself were the same. By using a GC/MS-SIM (selective ion monitoring) technique, the average value was found to be 0.51 ng/m(3) of air (range: 0.02 approximately 1.92 ng/m(3) of air). The trend of the residual levels in air particulates showed seasonal variation, increasing from autumn to winter and decreasing from winter to spring. The only exception was that the value in January was lower than those in December and February. Considering the content of BPA in the extract of the acidic fraction and the strength of the activities with the extract and BPA itself, the estrogenic activity due to BPA in the fraction seemed to decrease. In spite of this decline, the possibility remains that the estrogenic activity mainly originated from BPA. PMID- 15883674 TI - Effects of acid mine drainage and acidity on the activity of Choroterpes picteti (Ephemeroptera: Leptophlebiidae). AB - Survival and behavior of the mayfly Choroterpes picteti (Leptophlebiidae) exposed to acid mine drainage (AMD: pH 3.3-6.4) and a reservoir polluted with arsenic (pH 6.8) from Sao Domingos mine (Portugal) were studied in laboratory and in situ bioassays (48 h) with the Multispecies Freshwater Biomonitor, and compared with water from a reference river and acidified reference water (acid only). Metal body-burdens showed a negative pH dependency for Mn and As, a positive one for Pb, and for Zn, Cu, Co, and Cd a decrease at pH < 4.4. Generally, survival decreased with decreasing pH. The 48-h LC(50) (pH) for AMD and for acid only were similar (pH 4.8-4.9); however, the LT(20) (h) at pH 3.3 revealed AMD to be less toxic than acid only. C. picteti show diurnal rhythm with increased locomotor activity in the night. The circadian rhythm was weakened by acid exposure, but less so by AMD exposure. Compared to reference river water, ventilation was stimulated at pH < 6.0 in acid only and in reservoir water. Locomotion was stimulated at pH 5 in acid only and reservoir; however, it was reduced in all other treatments, when compared to reference river water. Under acid-only exposure, both locomotion and ventilation were significantly higher compared to AMD exposure at the corresponding pH values. The laboratory results were field validated. PMID- 15883675 TI - Toxicity of the crude oil water-soluble fraction and kaolin-adsorbed crude oil on Daphnia magna (Crustacea: Anomopoda). AB - Crude oil is a complex mixture of hydrocarbons entering aquatic environments from accidental or normal marine and transportation activities. Toxicologic crude oil analysis is usually performed on the basis of the water-soluble fraction. However, this yields only a partial estimate of the damage caused by these contaminants because a substantial hydrophobic amount can be adsorbed onto suspended solids (biotic and abiotic), which directly affects filter-feeding species and permits bioaccumulation through trophic relationships. This study determined the acute toxic damage sustained after 48 hours caused by seven types of crude oil from Tabasco, Mexico on the cladoceran Daphnia magna. Comparisons were documented based on the responses of D. magna from application of the water soluble fraction and exposure to entire crude oil samples adsorbed on kaolin clay. Oil-sorbed kaolin was more toxic than the water-soluble fraction in acute exposure. This confirms that tests of the water-soluble fraction tend to underestimate the toxic damage that can be produced in natural environments. Furthermore, chronic toxicity (21 days) was evaluated for crude oil samples adsorbed on kaolin at sublethal concentrations as established from Application Factors (AF) criteria. Results showed that in most cases, it is impossible to predict safe concentrations on the basis of LC(50) values because samples with lower acute toxicity exercised a greater influence on D. magna reproduction and survival when subjected to chronic exposure. PMID- 15883676 TI - The ratios of individual chemicals in a mixture determine the degree of joint effect: the climax hypothesis. AB - A new hypothesis, the Climax Hypothesis, is proposed to describe how the ratios of individual chemicals influence the joint effects of chemical mixtures. This hypothesis is derived from a generalized approach using "isobolograms". With this hypothesis, one can predict that for a given mixture, a curvilinear correlation exists between the joint effects and the ratios of individual chemicals and that this curve has a climax at the equitoxic ratio. This prediction is supported by the observed toxicity to Vibrio fischeri of 18 mixtures (12 binary mixtures, 4 ternary mixtures, and 2 quaternary mixtures). With this prediction, the Climax Hypothesis has some promising applications such as controlling the discharge of effluents in environmental science, maximizing detoxifying effects in medicine, and optimizing combined pesticides in agriculture. PMID- 15883677 TI - The development of marine Toxicity Identification Evaluation (TIE) procedures using the unicellular alga Nitzschia closterium. AB - Unicellular algae are highly sensitive to a wide range of toxicants and have been used extensively in ecotoxicological testing. This, along with their ability to grow in very small test volumes over short test durations, make them ideal test organisms for use in Toxicity Identification Evaluations (TIEs). Despite this, microalgae have not previously been used in marine TIE studies. In this study, the marine diatom Nitzschia closterium was shown to be a highly suitable test organism after modification of the standard test protocol to reduce test volumes to 6 mL and test duration to 48 h. The alga was tolerant to the chemicals used in phase I of the standard USEPA TIE protocol, and physical TIE manipulations had no effect on algal growth. The cation exchange procedure, however, inhibited algal growth, while the anion exchange procedure stimulated growth, making these two procedures unsuitable for use with this species. Of the buffers trialed for the graduated pH procedure, 0.01 M PIPES buffer was found to be suitable for buffering at pH 7 because it maintained the required pH over the duration of the test and did not affect the growth or sensitivity of the algae to one reference toxicant (copper). A trial TIE on a secondary-treated sewage effluent for discharge into coastal waters showed that the developed protocols could successfully be used to identify ammonia as the major toxicant in the effluent. PMID- 15883678 TI - Immune pathogenesis of multiple sclerosis. PMID- 15883679 TI - Optimisation of immunomodulatory therapies. PMID- 15883680 TI - Immunosuppressive therapy in multiple sclerosis. PMID- 15883681 TI - Autologous haematopoietic stem cell transplantation. PMID- 15883682 TI - Multiple sclerosis therapy: new strategies. PMID- 15883683 TI - Cellular therapy in multiple sclerosis: where we are and where we are going. PMID- 15883684 TI - Hypertension, antihypertensive treatment and stroke prevention. AB - Despite intensive research efforts, few effective treatments are available once stroke has occurred. Thus, stroke prevention should be a primary focus for all health care provides. The purpose of this article is to review the pathophysiological relevance of hypertension as a leading cause of stroke. This will be followed by an analysis of the evidence that blood pressure lowering with different antihypertensive drugs reduces the risk. Finally recent data suggest that the angiotensin II (ATII)-receptor antagonist losartan may offer advantages beyond blood pressure lowering, including attenuation of the central aortic reflected pressure wave, molecule-specific properties and neural protective influences on brain ATII type 2 receptors. PMID- 15883685 TI - Management of ischaemic stroke: statins and antiplatelet agents. PMID- 15883686 TI - Specific therapies for ischaemic stroke: rTPA and others. AB - In the last few years there have been several important advances in the understanding of cerebrovascular disorder pathophysiology that have impacted on stroke management. The development of timely and effective treatment strategies was and is still considered a high priority issue. Therapeutic options dramatically increased both in the prevention and overall in the treatment of acute ischaemic stroke (AIS). At present, whereas neuroprotection remains experimental, intravenous (i.v.) thrombolysis is the only specific therapy effective in reducing mortality and disability associated with stroke. The efficacy and safety of the antithrombotic therapy in AIS treatment are not well established, and few issues in clinical stroke management are more controversial. However, some studies have brought new light and new doubts on the roles of these traditional therapies. PMID- 15883687 TI - Early phase combined therapeutic management of acute ischaemic stroke. AB - An adequate treatment of ischaemic stroke in the early phase (28-48 h) is the most important factor for a better outcome. Thrombolysis with rTPA (within 3 h) and oral ASA 300 mg/days are the first therapeutic misures. Continuous monitoring of cardiological and haemodinamic parameters allows early detection of cardiac disturbances. Treatment of hypertension, low haematic oxigenation, hyperglicaemia, seizures and hypertermia is basic to improve outcome. Pharmacological therapy is only one of the components of effective multidisciplinary integrated management of ischaemic stroke; we remind also the precocity of rehabilitation procedures and an accurate psychological assessment. PMID- 15883688 TI - Antibodies associated with paraneoplastic neurological disorders. AB - Certain cancers, particularly small cell lung cancer, gynaecological or breast and also testicular, prostate, bowel and other cancers, can be associated with diverse and often heterogeneous neurological syndromes. In those cases with neurological syndromes, a proportion of patients will have a serum antibody to a defined antigen that is expressed both by the tumour and by the nervous system. The presence of the neurological syndrome often precedes the recognition of the tumour, and thus the detection of one of the specific antibodies leads to a search for the relevant tumour. In general, the antibodies are detected by immunohistochemistry or immunofluorescence on brain tissue. Their specificity can then be confirmed either by western blotting of neuronal or brain extracts, or by western blotting of the specific recombinant protein. There are also some ELISA assays available for detection of antibodies to specific antigens. PMID- 15883689 TI - Early invasive treatment (endarterectomy vs. stenting) of moderate-to-severe carotid stenosis in patients with transient ischaemic attack or minor stroke. AB - Carotid endarterectomy (CA) is an effective treatment for the secondary prevention of stroke in patients with carotid stenosis >50%. More recently, carotid angioplasty with stenting (CAS) has been introduced and found to be effective in case series and small clinical trials. Although CAS has been shown not to be inferior to CA, the comparative effects of early treatment (i.e., during the first month after transient ischaemic attack (TIA) or minor stroke) with these techniques are unknown. Early treatment is advocated, as recurrent stroke tends to present frequently in this time period. On this background, we designed a randomised clinical trial comparing the efficacy and safety of CA vs. CAS in patients who had suffered TIA or minor stroke in the antecedent month. The study design and methods of this multicentre pragmatic randomised parallel-group open trial are presented here. PMID- 15883690 TI - Early haemostatic therapy for spontaneous intracranial haemorrhage. AB - Intracranial haemorrhage (ICH) accounts for approximately 10%-15% of all strokes and is associated with the highest mortality rate and with the highest degree of disability among the survivors compared to all other strokes. The role of surgical and medical treatment is controversial. Recently, a parallel-group trial design between early surgery versus initial conservative treatment in patients with ICH showed no overall benefit from surgery. Several agents could theoretically be used. Activated recombinant factor VII administered within 4 hours after the onset of ICH seems to be the best candidate: in a recent study, it limited the growth of the haematoma, reduced mortality and improved functional outcome at 90 days. Further studies are necessary to define optimal dose of this drug, verify its efficacy and identify patients at high risk for thromboembolic complications. PMID- 15883691 TI - Local drug delivery in recurrent malignant gliomas. AB - In recurrent malignant gliomas, we scheduled a protocol by adding to systemic temozolomide a local treatment delivered through a reservoire positioned in the surgically created cavity, consisting of either mitoxantrone, liposome-loaded doxorubicine or nimustine (ACNU). The progression-free survival (PFS) and survival time (ST) of the whole group of 112 patients were 8.3 and 11 months, respectively, in GBM patients, and 14 and 18 months in AA patients. To limit the selection bias in recruitment we matched locally treated patients with the whole group of patients treated for 3 years and having undergone the same protocol with the exception of local drug delivery. Variables such as age, histology and local chemotherapy delivery were proved to be statistically significant independent factors on adjunctive PFS and ST. Another group of 12 recurrent malignant gliomas with further progression was locally managed according to convection-enhanced delivery (CED) of mitoxantrone; the preliminary results show good tolerability of the schedule. PMID- 15883692 TI - Brain tumoral epilepsy: a review. PMID- 15883693 TI - Cell therapies in neuro-oncology. AB - During their growth, malignant gliomas interact with the immune system and are able to escape immune reactions. Attempts to instruct the immune system to develop anti-glioma reactions have been partly unsuccessful. Recent advances in the molecular and cellular biology of dendritic cells (DC), however, may increase the chances of preparing effective "vaccines" against these tumours. We show that vaccination with DC pulsed with a tumour lysate considerably increases survival in mice bearing intracranial glioblastomas. These results support the development of DC-based clinical trials for patients with glioblastomas that do not respond to standard therapies. PMID- 15883694 TI - The role of the neurologist. AB - The contribution of the neurologist should focus on timely diagnosis with accurate differential diagnosis, indications for surgery and post-surgical treatment (in the setting of a multidisciplinary team), and follow-up of disease course/treatment-related complications. PMID- 15883695 TI - The interdisciplinary treatment of neuro-oncological patients: the role of the general oncologist. PMID- 15883696 TI - Beta-amyloid immunisation in Alzheimer's disease. PMID- 15883697 TI - A rehabilitative approach to the patient with brain cancer. AB - The aim of this essay is to describe our experience with patients affected by cerebral neoplasia within a diagnostic-clinical path based on a rehabilitative viewpoint. Our rehabilitative model is first of all an organisational process based on the patient's global management and on teamwork. It is also defined and qualified to be multidisciplinary. This model is in line with several studies published between 1998 and 2001 that led to recognition and management of the specific psychological problems associated with each stage of rehabilitation treatment. It was possible to individuate and prevent those environmental conditions that are dysfunctional to the psychological and cognitive processes not yet completely restored after a surgical trauma. PMID- 15883699 TI - Inflammatory neuropathies. PMID- 15883700 TI - Autoimmune muscular pathologies. AB - The T cell-mediated mechanism responsible for Polymyositis and inclusion Body Myositis and the complement-mediated microangiopathy associated with Dermatomyositis are reviewed. The management of autoimmune myopathies with the presently available immunotherapeutic agents as well as new therapies and ongoing trials are discussed. PMID- 15883701 TI - Neurological involvement in rheumatological diseases. AB - Rheumatological diseases can involve the central and the peripheral nervous system in many ways. Every structure-the brain, meninges, spinal cord, cranial nerves, peripheral nerves-can be affected. Early recognition of neurological abnormality can help achieving diagnosis of the underlying condition and prevent permanent sensorimotor or cognitive function loss. This review focuses on the clinical presentation of the neurological involvement in rheumatological diseases. PMID- 15883703 TI - Molecular pathogenesis of MEN2-associated tumors. AB - Although the gene responsible for multiple endocrine neoplasia type 2 (MEN2) was discovered many years ago, the exact mechanisms of tumor development in patients affected with RET germline mutations remain unknown. In vitro studies have certain pitfalls, one of which is the use of cell culture systems such as the NIH3T3 cells, in which RET usually is not expressed in contrast to the in vivo situation. Recent data suggest that an overrepresentation of mutant RET as a 'second hit' event might trigger tumorigenesis. However, alterations in other genes might contribute to this overrepresentation of RET or impact on MEN 2 related tumor development through completely different mechanisms and pathways. The final goal of further elucidating the natural history and pathogenesis of MEN2-related tumors should be the chance to offer patients with RET germline mutations an optimal cancer prevention (e.g. codon specific recommendations for prophylactic thyroidectomy) and treatment program, especially for metastatic medullary thyroid carcinoma for which presently no effective therapy other than surgery exists. PMID- 15883704 TI - SDHC mutations in hereditary paraganglioma/pheochromocytoma. AB - Mutations in genes coding for three of the four components of mitochondrial complex II can cause paragangliomas (PGLs)/pheochromocytomas. The three genes include SDHB, -C, and -D. SDHC and SDHD anchor the catalytic subunits SDHA and -B of mitochondrial complex II in the inner mitochondrial membrane. SDHD is maternally imprinted but SDHB and -C are not. While SDHD and -- to a lesser degree -- SDHB mutations have been found in many cases of hereditary PGL, SDHC mutations are rare. This article reviews the SDHC mutations described to date and discusses possible mechanisms of tumorigenesis. PMID- 15883705 TI - Pheochromocytoma in von Hippel-Lindau disease and neurofibromatosis type 1. AB - Clinical and genetic understanding of chromaffin tumors has been greatly enhanced in the last few years. Although some pheochromocytoma genes may still be unknown, the role of RET, VHL, SDHB, SDHD and NF1 genes is unequivocal and phenotypes are also being better characterized. The loss of function of VHL and NF1 genes can lead to a variety of tumors including phechromocytoma and their mechanism of action is under intensive investigation. Many different mutations are responsible for VHL gene inactivation but only missense mutations have been described so far in families with pheochromocytoma. Because of its large size extensive mutation analysis of the NF1 gene has seldom been performed, and mutations have only been identified in about 15% of patients. Several point mutations have been found in exon 31. Differences in pheochromocytoma phenotype in VHL or NF1 are not very pronounced, but it may be of some interest to consider the two groups separately. In VHL, pheochromocytoma has an earlier onset than in sporadic forms, it is often multiple, and malignancy is less frequent. The mean age of diagnosis is 28 years, the youngest patient being 5 years old. In NF1 patients pheochromocytoma phenotype is similar to sporadic forms. The mean age of pheochromocytoma onset is 42 years; 84% of patients have solitary adrenal tumors, 9.6% have bilateral adrenal disease and 6.1% have ectopic pheochromocytomas; malignant pheochromocytomas were identified in 11.5% of the cases. The group of pheochromocytoma susceptibility genes includes, along with the tumor suppressor genes VHL and NF1, the proto-oncogene RET and the genes encoding succinate dehydrogenase subunit D and succinate dehydrogenase subunit B. Whether there is a common pathway among these different genes is still a matter of debate. PMID- 15883706 TI - Pheochromocytoma-associated syndromes: genes, proteins and functions of RET, VHL and SDHx. AB - Pheochromocytoma are tumors derived from chromaffin cells that secrete catecholamines. These catecholamines may lead to increased blood pressure and even death. Historically, pheochromocytoma have been described as 10 tumor, i.e. about 10 were believed to be malignant, 10 were found to be extra-adrenal, and 10 were meant to be bilateral. Also, about 10 were considered to be hereditary. In these instances, they were most often part of either the multiple endocrine neoplasia type 2 (MEN 2) syndrome or the von Hippel-Lindau (VHL) disease. The genes (RET and VHL) involved have been known for several years and their function is the subject of ongoing investigation. Very recently, several genes (SDHD, SDHB, and SDHC) that belong to the mitochondrial complex II have been identified to be involved in the so-called pheochromocytoma-paraganglioma syndrome. Only SDHD and SDHB have so far been implicated in the pathogenesis of pheochromocytoma. PMID- 15883708 TI - Central nervous system manifestations in VHL: genetics, pathology and clinical phenotypic features. AB - This review focuses on CNS hemangioblastomas in von Hippel-Lindau (VHL) disease. The pathogenesis of these lesions remains unclear to date; however, biallelic inactivation of the VHL tumor suppressor gene is thought to be an important step. These benign tumors occur frequently in patients with VHL disease and produce symptoms by mass effect either by the tumor itself or an accompanying cyst or edema. Furthermore, cases of spontaneous hemorrhage have been described. Genetic testing for VHL germline mutations is recommended in all patients with hemangioblastoma and yearly screening, including MRI of the brain and spine, is recommended for all VHL disease patients. Treatment of these tumors is mainly surgical. In general, surgery is indicated in symptomatic hemangioblastomas and eventually also in asymptomatic tumors that exhibit radiographic progression. However, since most VHL disease patients harbor multiple lesions, a careful individual decision must be made in each case. The tumors can usually be completely removed by dissection in the plane between tumor and CNS tissue and coagulating and cutting of the numerous feeding vessels with low power. As long as consequent yearly surveillance is performed and lesions are adequately treated in time, the prognosis of CNS hemangioblastomas in VHL disease is good. Preoperative neurological deficit, however, will not improve after surgery in most patients. Local tumor recurrences are rare. PMID- 15883707 TI - Multiple endocrine neoplasia type 2. AB - Multiple endocrine neoplasia type 2 (MEN 2) is an autosomal dominant inherited cancer syndrome. Predisposition to MEN 2 is caused by germline mutations of the RET proto-oncogene on chromosome 10q11.2 [1]. There are three clinically distinct forms of MEN 2 syndrome -- MEN 2A, familial medullary thyroid carcinoma (FMTC), and MEN 2B. In all of these subtypes, medullary thyroid carcinoma (MTC) is a key. MEN 2A is the most common subtype of MEN 2. Clinical features of the MEN 2A syndrome include medullary thyroid carcinoma (MTC) and/or C-cell hyperplasia (CCH) in almost all affected individuals, pheochromocytoma (approximately 50%) and hyperparathyroidism (HPT) (15-30%). MEN type 2B is the most aggressive of the MEN 2 variants and accounts for approximately 5% of all cases of MEN 2. MEN 2B is similar to MEN 2A but is characterized by the earlier onset of the disease and by developmental abnormalities. In FMTC, the third form of MEN 2, MTC is the only clinical feature. Introduced in recent years and still developing genetic testing of individuals at highest hereditary risk of MEN 2 syndrome holds the possibility of early detection and improved treatment and prognosis. PMID- 15883709 TI - Ophthalmological manifestations in VHL and NF 1: pathological and diagnostic implications. AB - Von Hippel-Lindau disease (VHL) and neurofibromatosis type 1 (NF 1) are hereditary multitumor syndromes that show associated ocular manifestations. Capillary retinal angioma, a benign vascular tumor, is the classical ocular lesion in VHL. It often appears as the first manifestation of the disease and may thus lead to the diagnosis of VHL. Since small angiomas can be treated easily by laser photocoagulation, a regular ocular screening of VHL patients is recommended. Ocular manifestations of NF 1 are more diverse as compared to VHL. Lisch nodules of the iris are an important diagnostic criteria of NF 1 since they can be found in almost every affected patient. Optic glioma can occur both intraorbitally and intracranially. The intraorbital form causes progressive protrusion of the globe and eventually blindness. Extension of the tumor beyond the chiasm worsens the prognosis quoad vitam. The hallmark of NF 1, namely cutaneous neurofibroma can cause visual impairment when affecting the skin of the eyelids. The rare intraorbital pexiform neurofibroma is associated with abnormal development of the orbital bones and infantile glaucoma. It may result in orbital mass effects and therefore may need surgical excision. PMID- 15883710 TI - Mutations of the SDHB and SDHD genes. AB - The succinate dehydrogenase (SDH) is a mitochondrial enzyme complex with an important role in oxydative phosphorylation and intracellular oxygene sensing and signaling. Mutations in the SDHB (1p35-36) and SDHD subunits (11q23) give rise to the paraganglioma syndromes (PGL), namely PGL 4 and PGL 1, and generate paraganglioma and pheochromocytoma. For both genes mutations have been described that result in a loss of function of the gene products. SDHBmutations were found in five of eight exons and in two introns, SDHD mutations in all four exons and one intron. Phenotypes and rate of malignancy of SDHB and SDHD seem to be different, with a higher frequency of head-and-neck tumors in SDHD and indications of a higher risk of malignancy in SDHB mutations. As routine diagnostic procedure all SDH mutation carriers should have urine catecholamine analysis as well as pelvic, abdominal, thoracic and skull/neck MRI. PMID- 15883711 TI - Paragangliomas of the head and neck: diagnosis and treatment. AB - Paragangliomas of the head and neck (HNP) represent rare tumors of neural crest origin. They are highly vascular neoplasms that are benign in the majority of cases. The site of origin defines the name given those tumors. In the head and neck, they most commonly occur at the carotid bifurcation, where they are referred to as carotid body tumors (CBT). Other common sites of origin are the jugular bulb (jugular paraganglioma; JP), the tympanic plexus on the promontory (tympanic paraganglioma; TP) and the vagal nerve (vagal paraganglioma; VP). Patients with cervical paragangliomas frequently present with a painless, slowly enlarging mass in the lateral neck. In many patients with TP and JP, tinnitus and hearing loss are early symptoms. JP patients often suffer from lower cranial nerve deficits. Evaluation by an imaging modality is necessary to establish the diagnosis. Imaging procedures frequently used include B-mode sonography with color-coded Doppler sonography, computed tomography (CT), magnetic resonance imaging (MRI) and digital substraction angiography (DSA). Debate exists in the literature regarding the different treatment modalities for paragangliomas which include surgery, radiotherapy and stereotactic radiosurgery. The role of preoperative angiography and embolization has also been a matter of discussion. The diagnostic work up and the different treatment options for patients with head and neck paragangliomas will be presented and discussed. PMID- 15883712 TI - Imaging of pheochromocytoma and paraganglioma. AB - Paragangliomas are tumours that arise within the sympathetic nervous system originating from the neural crest. These tumours can be found anywhere from the neck to the pelvis in locations of sympathetic ganglions. Although in the majority of paragangliomas the diagnosis is based on measuring catecholamines and metabolites in plasma or urine, imaging plays an important preoperative role. Today, there are several morphological and radionuclide imaging methods available that predict tumour localisation and tumour extent and give anatomic information to the surgeon. MRI is the morphological imaging modality of choice in localising pheochromocytomas and extra-adrenal paragangliomas. It provides excellent anatomic detail and has the advantage of lacking ionising radiation. The overall accuracy of computed tomography (CT) in detecting primary adrenal pheochromocytomas is very high, but CT lacks in specificity as difficulties may occur in distinguishing between paragangliomas and other tumour entities. The major advantages of radionuclide imaging are very high specificity and routinely performed whole-body scanning. Furthermore, metabolic imaging is not influenced by artifacts like scar tissue or metallic clips in post-surgical follow-up. Currently, a reported specificity of 99% and a cumulative sensitivity of about 90% in paragangliomas make (123)I-MIBG the most important nuclear imaging method. However, (18)F-DOPA-PET seems to be a very promising procedure which offers higher accuracy. The higher spatial resolution of PET-scanners enables the detection of small lesions not visualised with (123)I-MIBG. Both use of radiolabelled somatostatin analogue like (111)In-pentetreotide and (18)F-FDG is limited due to low specificity of the tracers and should be restricted to MIBG- and F-DOPA-negative cases. PMID- 15883714 TI - Molecular subtyping of clinical isolates of Candida albicans and identification of Candida dubliniensis Malaysia. AB - The genotypes of 221 recent isolates of Candida albicans from various clinical specimens of 213 patients admitted to the University Malaya Medical Centre, Malaysia was determined based on the amplification of a transposable intron region in the 25 S rRNA gene. The analyses of 178 C. albicans isolated from nonsterile clinical specimens showed that they could be classified into three genotypes: genotype A (138 isolates), genotype B (38 isolates) and genotype C (2 isolates). The genotyping of 43 clinical isolates from sterile specimens showed that they belonged to genotype A (29 isolates), genotype B (10 isolates), genotype C (2 isolates) and genotype D (2 isolates). The overall distribution of C. albicans genotypes in sterile and nonsterile specimens appeared similar, with genotype A being the most predominant type. This study reported the identification of C. dubliniensis (genotype D) in 2 HIV-negative patients with systemic candidiasis, which were missed by the routine mycological procedure. The study demonstrated the genetic diversity of clinical isolates of C. albicans in Malaysia. PMID- 15883715 TI - Renal ultrasonography and detection of pseudomycelium in urine as means of diagnosis of renal fungus balls in neonates. AB - OBJECTIVE: To present a series of neonates with renal fungus balls diagnosed by ultrasonography, urine culture and/or by the detection of Candida pseudomycelium in urine. PATIENTS AND METHODS: We revised the clinical records of neonates for whom the diagnosis of renal fungus ball was established by ultrasound and laboratory studies; these patients had been hospitalized at the National Institute of Pediatrics in Mexico between January 1st, 1999 and December 31st, 2002. RESULTS: During the study period, 9 neonates were diagnosed with renal fungus ball. In 7 cases, the ethiologic agent was Candida albicans; whereas it was C. tropicalis in one case and C. parapsilosisin the other. Urine culture was positive (> or =10,000 UFC/ml) in 8 cases, whereas the fungal density was only 2400 UFC/ml in the last sample. Pseudohyphae were present in all cases and ultrasonography showed fungus ball in every case. All patients received a single antifungal drug, either amphotericin B or fluconazole. All the patients recovered and none of them required surgical treatment. Control postreatment by ultrasound studies showed that the fungus balls had disappeared in every case. CONCLUSION: The diagnosis of Candida renal fungus balls based on the ultrasound study and urine culture is also substantiated by the detection of pseudomycelium in the centrifugation pellet of urine samples, which is a fast diagnostic method. This approach permitted an early diagnosis and treatment of Candida renal fungus balls. PMID- 15883716 TI - The inhibition of Candida albicans by selected essential oils and their major components. AB - Many volatile oils are known to possess antifungal properties and are potentially applicable as antimycotic agents. By studying the efficacy of essential oils against different pathogenic mycetes, we have evaluated the in-vitro inhibiting activity of some essential oils and their main constituents against a strain of Candida albicans. Sixteen commercial essential oils and forty-two pure constituents (alcohols, aldehydes, ketons, phenols and hydrocarbons), were tested by using a semisolid agar antifungal susceptibility (SAAS) method. Gas chromatography/mass spectroscopy analyses of the oils tested were performed. The essential oils of Origanum vulgare, Satureja montana, Mentha piperita, Cinnamomum verum, Cymbopogon flexuosus showed maximum inhibitory activity (MIC = 500 ppm) after 7 days. According to the results of the examination of pure constituents, beta-phellandrene proved to be the most interesting component among cyclic monoterpenic hydrocarbons as it showed a strong activity (MIC = 50 ppm). The most active of phenols was carvacrol (MIC 100 ppm). The open-chain alcohol 1-decanol was the most active of alcohols at 50 ppm. Finally, among aldehydes, a strong activity was shown by trans-cynnamaldehyde (MIC 50 ppm). PMID- 15883717 TI - A prospective epidemiologic survey on the prevalence of onychomycosis and dermatophytosis in male boarding school residents. AB - Dermatophyte infections and onychomycosis are not usually serious in term of mortality; however, they may have significant clinical consequences such as secondary bacterial infections, chronicity, therapeutic difficulties and esthetic disfigurement in addition to serving as a reservoir of infection. Our aim was to determine the prevalence of onychomycosis and dermatophytosis in a selected high risk group, consisting of male boarding school residents. A total of 410 males inhabiting two houses were evaluated by two dermatologists. In cases of clinical suspicion, appropriate samples were taken for direct microscopy and culture. The results showed that the prevalences of tinea pedis (athlete's foot) and pure pedal onychomycosis were 51.5% (n:211) and 4.4% (n:18), respectively. Thirty cases of those with tinea pedis were complicated by toenail onychomycosis. Tinea cruris was present only in five cases with tinea pedis. Interestingly 71.1% of those with tinea pedis and 45.8% of those with onychomycosis, associated with or without tinea pedis were unaware of their diseases. The most common fungal isolate was Trichophyton rubrum (76.6%) followed by Epidermophyton floccosum (11.6%), T. interdigitale (10.55%). Approximately one third of the cultures from nail specimens yielded pure growths of nondermatophyte moulds or Candida albicans. In conclusion, we found unexpectedly high prevalences of occult athlete's foot and toenail onychomycosis among the male residents of student houses. Our results indicate that health-care workers of such common boarding houses should be more aware of clinical and subclinical dermatophyte infections and onychomycosis, and have more active approaches to educational measures and management strategies to prevent further infections. To our knowledge, this is the first epidemiologic study on the prevalences of dermatophytosis and onychomycosis in boarding-houses from Turkey. PMID- 15883718 TI - The use of new probes and stains for improved assessment of cell viability and extracellular polymeric substances in Candida albicans biofilms. AB - Phenotypic and genotypic cell differentiation is considered an important feature that confers enhanced antifungal resistance in candidal biofilms. Particular emphasis has been placed in this context on the viability of biofilm subpopulations, and their heterogeneity with regard to the production of extracellular polymeric substances (EPS). We therefore assessed the utility of two different labeled lectins Erythrina cristagalli (ECA) and Canavalia ensiformis (ConA), for EPS visualization. To evaluate the viability of candidal biofilms, we further studied combination stains, SYTO9 and propidium iodide (PI). The latter combination has been successfully used to assess bacterial, but not fungal, viability although PI alone has been previously used to stain nuclei in fungal cells. Candida albicans biofilms were developed in a rotating disc biofilm reactor and observed in situ using confocal scanning laser microscopy (CSLM). Our data indicate that SYTO9 and PI are reliable vital stains that may be used to investigate C. albicans biofilms. When used together with ConA, the lectin ECA optimized EPS visualization and revealed differential production of this material in mature candidal biofilms. The foregoing probes and stains and the methodology described should help better characterize C. albicans biofilms in terms of cell their viability, and EPS production. PMID- 15883719 TI - Characterization of clinical isolates of pathogenic Nocardia strains and related actinomycetes in Thailand from 1996 to 2003. AB - In Thailand from 1996 to 2003, 171 strains of pathogenic aerobic actinomycetes from clinical specimens were isolated. Of those strains, 134 were mycolic acid containing actinomycetes, including 96 strains of Nocardia species. Others included 10 strains of Gordonia, 14 strains of Rhodococcus, and 22 strains of Mycobacterium. One strain each of the genera Tsukamurella and Corynebacterium were also isolated. Also identified were 27 strains of non-mycolic acid containing actinomycetes. Our identification studies of 96 strains of Nocardia species showed that significant pathogens in Thailand were N. beijingensis (18 strains), N. cyriacigeorgica (13 strains), and N. farcinica (34 strains); the most prevalent species was N. farcinica (35.4%). We also isolated four strains of N. asiatica, five strains of N. asteroides sensu stricto, four strains of N. nova, seven strains of N. otitidiscaviarum, eight strains of N. transvalensis, and two strains of N. pseudobrasiliensis. PMID- 15883720 TI - The first clinical isolates of Candida dubliniensis in Slovakia. AB - Candida dubliniensis, yeast closely related to Candida albicans, is a new pathogen associated mainly with infections of immunocompromised hosts. In this study, we report the first isolation of three isolates of C. dubliniensis in Slovakia. The first selection of both C. albicans and C. dubliniensis from the other Candida species was done on the basis of specific green color of primoculture grown on CHROMagar Candida. The presumptive identification was completed by supplemental tests: germ-tube formation, production of chlamydospores, ability or inability to grow at 42 and 45, degrees C and by commercial set API 20C AUX. Parallely, the discrimination between both species was performed by PCR assay using primers specific for Candida dubliniensis. PMID- 15883721 TI - Simultaneous chronic pulmonary paracoccidiodomycosis and disseminated cryptococcosis in a non-HIV patient. AB - An unusual case of chronic pulmonary paracoccidiodomycosis and disseminated cryptococcosis in a non-HIV infected patient is reported in a 72-year-old previously healthy man. A chest radiograph disclosed a bilateral diffused interstitial infiltrate involving middle and lower lung fields. Specimen samples taken from the tracheal tube revealed yeast-like organisms suggestive of Cryptococcus neoformans and Paracoccidiodes brasiliensis. Blood and cerebrospinal fluid culture was positive for C. neoformans and the immunodiffusion test against paracoccidiodin revealed a precipitation band. The patient died 24 days after the admission. PMID- 15883722 TI - In vitro susceptibility testing of Microsporum gypseum isolated from healthy cattle and soil samples against itraconazole, terbinafine, fluconazole and topical veterinarian drugs. AB - The present study evaluated in vitro susceptibility testing of dermatophytes isolates from healthy cattle and soil samples against three antifungal agents and three topical veterinarian drugs. Itraconazole and terbinafine showed a higher in vitro fungicidal activity than fluconazole. The veterinarian drugs LEPECID and iodine 5% were more active in vitro than the UNGUENTO spray. All drugs showed fungicidal activity against Microsporum gypseum, and they may be considered as efficient agents for the topical treatment of dermatophytoses in cattle. PMID- 15883723 TI - Effects of carbon and nitrogen sources, carbon-to-nitrogen ratio, and initial pH on the growth of nematophagous fungus Pochonia chlamydosporia in liquid culture. AB - The effects of carbon and nitrogen sources, carbon-to-nitrogen ratio (C:N) and initial pH value on the growth and sporulation of the nematophagous fungus Pochonia chlamydosporia in liquid culture were examined. Among the 21 carbon sources and 15 nitrogen compounds tested, the optimal carbon and nitrogen sources for mycelial growth were sweet potato and L: -tyrosine, and for sporulation were sweet potato and casein peptone. A C:N ratio of 10:1 at pH 3.7 gave the maximum yield of conidia and a C:N ratio of 40:1 at pH 6.8 gave the maximum biomass. The initial pH value had a significant effect on mycelial growth and conidial production, with the optimal ranges being 3.5-4.5 for sporulation and 5-6 for growth. Maximum conidial production was obtained at an initial pH of 4.0 and the maximum biomass at pH 6.0. The results also showed that the final pH after 7 days cultivation was always higher than the initial value. The variability in growth and sporulation of seven strains of P. chlamydosporia in liquid culture was also compared and discussed. PMID- 15883724 TI - Isolation of Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) from the Chagas disease vector, Triatoma infestans (Hemiptera: Reduviidae) in Argentina. AB - A survey for natural entomopathogenic fungi of the Chagas disease vector Triatoma infestans was conducted in five provinces of Argentina since 2001. Nymphs (1.5%) and adults (3.3%) infected with a strain of the fungus Beauveria bassiana were found at Dean Funes, Cordoba province, Argentina. Field collected insects that died in the laboratory were maintained in moist chambers and incubated at 22 degrees C. Beauveria bassiana from infected insects was cultured on SDAY media. Pathogenicity tests were conducted with a conidial suspension (1 x 10(7) conidia/ml) of this isolate on T. infestans adults. A mortality rate of 100% was obtained at 15 days post-infection. This is the first record of natural infection of T. infestans by B. bassiana. PMID- 15883725 TI - Induction of apoptosis by fungal culture materials containing cyclopiazonic acid and T-2 toxin in primary lymphoid organs of broiler chickens. AB - Thirty-six, twenty-eight-day-old broiler chicks were randomly distributed into three groups of 12 birds each. Two groups were fed diets containing 10 ppm cyclopiazonic acid (CPA) and 1ppm T-2 toxin, respectively, to determine the mechanism of cell death in spleen and thymus at 6, 12, 24, and 36 h of post treatment. The other group served as control. T-2 toxin treated group showed significant (P < 0.01) induction of apoptosis in thymus with peak induction at 24 h post-treatment where as, no significant differences were observed between the control and CPA groups. The CPA toxin treated group showed significant (P < 0.01) induction of apoptosis in spleen with peak induction at 24 h post-treatment. No significant differences were observed between the control and T-2 toxin group even though the latter showed a slight increase in the quantity of apoptotic cells at 36 h post-treatment in spleen. The semi-thin sections stained with toluidine blue from the spleen of CPA treated group exhibited crescent margination of chromatin against the nuclear envelope and shrinkage of lymphoid cells without any surrounding inflammation, the characteristics of apoptosis. The apoptotic thymocytes from T-2 fed birds appeared shrunken with condensed nucleus and showed crescent margination of chromatin against the nuclear envelope without any surrounding inflammation when compared with well-defined nuclei with dispersed chromatin in normal thymocytes. Ultrastructurally, splenocytes of the CPA treated group and thymocytes of the T-2 toxin treated birds showed apoptotic bodies characterized by crescent margination of the chromatin against the nuclear envelope. The study indicates that one route of the CPA and T-2 toxin induced cell death in lymphoid organs of broiler chicken is by apoptosis. PMID- 15883726 TI - Fusarium species of the Gibberella fujikuroi complex and fumonisin contamination of pearl millet and corn in Georgia, USA. AB - This study was designed to identify and compare the Fusarium species of the Gibberella fujikuroi complex on pearl millet (Pennisetum glaucum (L.) R. Br) and corn (Zea mays L.) crops grown in southern Georgia, and to determine their influence on potential fumonisin production. Pearl millet and corn samples were collected in Georgia in 1996, 1997 and 1998. Three percent of the pearl millet seeds had fungi similar to the Fusarium species of the G. fujikuroi species complex. One hundred and nineteen representative isolates visually similar to the G. fujikuroi species complex from pearl millet were paired with mating population A (Fusarium verticillioides (Sacc.) Nirenberg), mating population D (F. proliferatum (Matsushima) Nirenberg) and mating population F (F. thapsinum (Klittich, Leslie, Nelson and Marasas) tester strains. Successful crosses were obtained with 50.4%, 10.1% and 0.0% of these isolates with the A, D and F tester strains, while 39.5 of the isolates did not form perithecia with any tester strains. Two of the typical infertile isolates were characterized by DNA sequence comparisons and were identified as Fusarium pseudonygamai (Nirenberg and O'Donnell), which is the first known isolation of this species in the United States. Based on the pattern of cross-compatibility, conidiogenesis, colony characteristics and media pigmentation, a majority of the infertile isolates belong to this species. Fumonisins FB(1) and FB(2) were not detected in any of the 81 pearl millet samples analyzed. The species of the G. fujikuroi species complex were dominant in corn and were isolated from 84%, 74% and 65% of the seed in 1996, 1997 and 1998, respectively. Representative species of the G. fujikuroi species complex were isolated from 1996 to 1998 Georgia corn survey (162, 104 and 111 isolates, respectively) and tested for mating compatibility. The incidence of isolates belonging to mating population A (F. verticillioides) ranged from 70.2% to 89.5%. Corn survey samples were assayed for fumonisins, and 63% to 91% of the 1996, 1997 and 1998 samples were contaminated. The total amount of fumonisins in the corn samples ranged from 0.6 to 33.3 microg/g. PMID- 15883727 TI - Co-occurrence of aflatoxin B(1) and cyclopiazonic acid in sour lime (Citrus aurantifolia Swingle) during post-harvest pathogenesis by Aspergillus flavus. AB - During hot and humid seasons, extensive rot of sour lime was observed to be caused by Aspergillus flavus. In view of this, investigations were undertaken to obtain data on the production of various toxins by A. flavus during post harvest pathogenesis of sour lime. Sixty percent of the pathogenic A. flavus isolates were detected to be aflatoxin B(1) producers in sour lime tissue. It was also noted that thirty three percent of aflatoxigenic A. flavus isolates had the potential to coproduce cyclopiazonic acid (CPA). Such aflatoxigenic isolates produced quantitatively more CPA (ranging from 250.0 to 2501.3 microg/kg) than aflatoxin B(1) (ranging from 141.3 to 811.7 microg/kg) in the affected sour lime. This study demonstrates for the first time that sour lime are a favourable substrate for aflatoxin B(1) and cyclopiazonic acid production by A. flavus isolates. This is of great concern to the health of consumers. PMID- 15883728 TI - Comparison of in vitro cytotoxicity of Fusarium mycotoxins,deoxynivalenol, T-2 toxin and zearalenone on selected human epithelial cell lines. AB - Three human epithelial cell lines (CaCo-2, HEp-2 and HeLa) implicated as potential targets for three Fusarium toxins were tested for the extent of survival on exposure to increasing toxin concentration and incubation periods. Cytotoxicity assay using 3(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) was carried out with deoxynivalenol (DON), T-2 toxins and zearalenone (ZON) on CaCo-2, HEp-2 and HeLa cell lines. Of the three cell lines used, HeLa was the most sensitive, eliciting cell death after 2 days exposure at 100 ng ml(-1)with T-2 toxin. HeLa was the only cell line to exhibit cytotoxicity towards ZON showing cell death at 1000 ng ml(-1)after 2 days which increased to 4 days, showing substantial cell death at 200 ng ml(-1). HEp-2 was sensitive to DON showing cell death after 2 days (100 ng ml(-1)) with complete cell death occurring at 200 ng ml(-1) after 4 days of exposure. Substantial cytoxicity of T 2 towards HEp-2 occurred after 2 days at 1000 ng ml(-1) and complete cell death occurred with 100 ng ml(-1) at day 4. The CaCo-2 cell line was generally resistant to the mycotoxins tested between 100 and 1000 ng ml(-1). This study shows that cytotoxicity of Fusarium toxins to epithelium cell lines is concentration- and time- dependant and results from ZON-HeLa interaction indicate possible cell type-mycotoxin specificity. PMID- 15883729 TI - Penicillium verrucosum occurrence and ochratoxin A contents in organically cultivated grain with special reference to ancient wheat types and drying practice. AB - This study addresses the relationship between the ochratoxigenic strains of Penicillium verrucosum and ochratoxin A (OTA) contents in organically cultivated grain. It included 37 combined, non-dried grain samples from farmers with no drying facilities as well as 19 non-dried and 22 dried samples from six farms with on-farm drying facilities (Case studies 1-6). The study focused on the ancient wheat type spelt but also included samples of wheat, rye, barley, oats, triticale, emmer, and einkorn. All 78 samples were analysed for moisture content (MC) and occurrence of P. verrucosum. The latter was assessed by plating non disinfected kernels on DYSG agar and counting those contaminated by the fungus. Fifty-five samples were analysed for OTA. Most of the combine harvested samples (82%) were contaminated with P. verrucosum prior to drying. This was ascribed to difficult harvest conditions and many samples of spelt, which was significantly more contaminated by P. verrucosum than oats, wheat and barley. Though not statistically significant, the results also indicated that spelt was more contaminated than rye, which is usually regarded the most sensitive small grain cereal. No correlation was found between number of kernels contaminated by P. verrucosum and OTA content. Despite many non-dried samples being contaminated by P. verrucosum, only two exceeded the EU maximum limit for grain (5 ng OTA g(-1)), both being spring spelt with 18 and 92 ng g(-1), respectively. The problems were most likely correlated to a late harvest and high MC of the grain. The case studies showed exceedings of the maximum limit in a batch of dried oats and spring wheat, respectively, probably to be explained by insufficient drying of late harvested grain with high MC. Furthermore, our results clearly indicate that OTA is not produced in significant amounts in samples with MCs below 17%. All dried samples with MCs above 18% exceeded the 5 ng OTA g(-1) limit in grain. However, no correlation between MC and the amount of OTA produced was found. PMID- 15883730 TI - Morphology and phylogeny of Botryosphaeria dothidea causing fruit rot of olives. AB - The taxonomic position of the causal agent of fruit rot of olives was determined from fresh collections of the fungus from central Greece. In culture it formed two types of conidia, namely fusiform, hyaline, aseptate conidia typical of the genus Fusicoccum, and dark-walled, ovoid, ellipsoid or fusiform, 1-2 septate conidia that are not typically observed in Fusicoccum. A phylogenetic analysis based on ITS and EF1-alpha sequences placed the fungus within the same clade as Fusicoccum aesculi, which is the anamorph of Botryosphaeria dothidea, and the type of the genus Fusicoccum. PMID- 15883731 TI - Characterization and in vitro expression patterns of extracellular degradative enzymes from non-pathogenic binucleate Rhizoctonia AG-G. AB - Many filamentous fungi produce an array of extracellular enzymes that acting in cell walls release elicitors of the plant defense response These enzymes may therefore be important in biocontrol applications. The aim of this study was to characterize extracellular degradative enzymes produced by a non-pathogenic binucleate isolate of Rhizoctonia AG-G. The fungus was grown in liquid culture supplemented with pectin, polygalacturonic acid or glucose as a carbon sources and filtrates of the culture media were analyzed for the detection of pectinolytic and glucan hydrolytic enzymes. Using only pectin as a carbon source, secretion of polygalacturonases and methylesterases was found. When the liquid medium was supplemented with polygalacturonic acid, only polygalacturonase activity was detected. However, when glucose was used as carbon source beta-1,3 and beta-1,6 glucanases activities were detected, using laminarin and pustulan as substrates, but none of the pectinolytic activities were found. These enzymes were partially purified and characterized. The beta-(1,3)(1,6) glucanase and polygalacturonase enzymes showed to be active against cell wall polysaccharides from potato sprouts. These enzymes may have an important role in fungus-plant cell wall interaction. This is the first study about the production of extracellular enzymes by non-pathogenic binucleate Rhizoctonia AG-G. PMID- 15883732 TI - Virulence and molecular diversity in Colletotrichum graminicola from Brazil. AB - Genetic diversity among 37 isolates of the sorghum anthracnose pathogen Colletotrichum graminicola, from four geographically distinct regions of Brazil, was evaluated by RAPD and RFLP-PCR markers and virulence characters on a set of 10 differential sorghum genotypes. Twenty-two races were identified and race 13B was the most frequent, but present in only two regions. RAPD analysis revealed 143 polymorphic bands that grouped the isolates according to their geographic origin, but not by their virulence phenotypes. RFLP with HaeIII, MspI, HinfI, HhaI, HpaII, EcoRI, HindIII, PstI, RsaI, Taq alphaI, and AluI enzymes over ITS domains and 5.8 rDNA genes of C. graminicola did not show differences among the isolates, indicating high conservation of these restriction sites. Molecular polymorphism was observed among isolates belonging to the same race. No association between virulence phenotypes and molecular profiles was observed. PMID- 15883733 TI - Assessment of laboratory methods for evaluating cassava genotypes for resistance to root rot disease. AB - Field evaluation of six cassava genotypes for resistance to root rot disease was compared with three rapid laboratory methods (whole root inoculation, root slice inoculation, and stem inoculation) for resistance screening. Both the field evaluation and the three laboratory methods separated the varieties into resistant and susceptible groups. Genotypes 30572 and 91/02324 were resistant while 92/0247, 92/0057 and TME-1 were susceptible. One genotype (30001) was not consistent in its reaction between field evaluation and laboratory assays. In the laboratory assays with three fungal pathogens, different pathogens varied in their levels of virulence on host genotypes. With the most virulent pathogen (Botryodiplodia theobromae), the majority of the genotypes reacted in the same way across trials with the root slice and whole root assays. Due to the good correlation between the whole root assay and the field results, we recommend this for the routine assessment of cassava resistance to root rot disease and for the analysis of virulence of pathogen isolates. However, because of the advantages in terms of economy of labour, space, time, quantity of root and inoculum required, the root slice assay could be used for the preliminary screening of large cassava accessions. The selected genotypes can then be further screened with the whole root inoculation method. PMID- 15883734 TI - Ergosterol from Phytophthora drechsleri, a unusual metabolite of a member of this genus. AB - Ergosterol was isolated from the plant pathogenic pseudofungus Phytophthora drechsleri grown on clarified V8 booth (CV8-B). Its structure was confirmed by comparison to an authentic sample. The species was identified by morphological analysis and molecular characterization by PCR: ITS (Internal transcribed spaces). This is the first report of this sterol in Phytophthora. This result is unusual because Phytophthora fungi were previously thought to be unable to synthesize sterols and the Oomycetes in general do not produce ergosterol. PMID- 15883736 TI - The COX-2 inhibitor, rofecoxib, ameliorates dextran sulphate sodium induced colitis in mice. AB - OBJECTIVE: We have evaluated the efficacy of the selective cyclo-oxygenase (COX) 2 inhibitor, rofecoxib, for the prevention of experimental colitis. MATERIAL AND METHODS: To induce colitis BALB/c mice received 5% dextran sulphate sodium (DSS) in their drinking water continuously for 7 days. Rofecoxib (2.5-10 mg/kg body weight, p.o.) was administered throughout the treatment period with DSS. Colitis was quantified by a clinical damage score, colon length, weight loss, stool consistency and rectal bleeding. Inflammatory response was assessed by neutrophil infiltration, determined by histology and myeloperoxidase (MPO) activity. Interleukin (IL)-1beta, prostaglandin (PG)E2 and PGD2 levels in colon mucosa and the immunohistochemical expression of COX-1 and -2 were also studied. RESULTS: The COX-2 inhibitor ameliorated severe colitis, reduced the degree of inflammation through reduction of neutrophil infiltration and IL-1beta levels. PGE2, and PGD2 synthesis were significantly reduced in DSS-treated groups. Indeed, treatment with rofecoxib diminished the lost of COX-1 caused by DSS in the crypt epithelium whereas expression of COX-2 remained unaffected. CONCLUSIONS: Rofecoxib is protective in acute DSS-induced colitis, probably by reducing neutrophil infiltration, inhibiting up-regulation of IL-1beta and returning to normal COX-1 expression in the inflamed colonic mucosa. PMID- 15883737 TI - Esophageal ATP synthase and keratinocyte growth factor gene expression changes after acid and bile-induced mucosal damage. AB - OBJECTIVE AND DESIGN: Intramural gene expression changes may be critically involved in tissue damage, defense and repair after esophageal regurgitation. The aims were to characterize the consequences of short-term exposure to luminal bile, acid, or bile mixed with acid on the beta-ATPase, keratinocyte growth factor 1 (KGF-1) and KGF receptor (KGF-R) expressions within the mucosa and the muscle layer in a large animal model. MATERIALS AND SUBJECTS: Esophageal segments of anesthetized dogs were exposed to saline (n = 3), diluted canine bile (n = 6), hydrochloric acid (n = 5) or bile + hydrochloric acid (n = 5), and tissue biopsies were taken at the end of the 180-min observation period. Semiquantitative reverse transcriptase polymerase chain reactions were carried out and the degree of histological damage was evaluated on the 0-16-grade Geisinger scoring scale. RESULTS: Acid exposure was followed by a significant decrease in the level of beta-ATPase expression in the mucosa, and parallel increases in KGF-1 and KGF-R expression. Corresponding changes in the muscle layer were not significant. Bile alone evoked more severe tissue damage, with significantly decreased beta-ATPase levels in both the mucosa and the muscle, whereas the KGF-1 expression did not change significantly. The bile + acid treatment induced an intermediate state, with significant beta-ATPase transcription level decreases in both layers, while the mucosal KGF-1 expression was lower than that following acid treatment alone. CONCLUSIONS: The acid-induced transcriptional level downregulation of mucosal beta-ATPase gene expression in the smooth muscle layer was exacerbated by bile, but the concomitant KGF and KGF R gene expression changes may indicate the start of a consecutive repair process. PMID- 15883738 TI - Effect of resveratrol in experimental osteoarthritis in rabbits. AB - OBJECTIVE: Resveratrol (trans-3,4',5-trihydroxystilbene) is a phytoalexin found in high concentration in the skins of grapes and red wines which has been shown to have antiinflammatory, anticancerogen and antioxidant properties. Resveratrol is a potent and specific inhibitor of nuclear factor kappa B (NF-kappaB). Resveratrol also inhibits COX-2 gene expression and enzyme activity. We aimed to determine the in vivo effects of intra-articular injections of resveratrol on cartilage and synovium in an experimental osteoarthritis (OA) model in rabbits. METHODS: As OA model, rabbits underwent unilateral anterior cruciate ligament transection (ACLT). Five weeks after test group was injected with 10 microMol/kg resveratrol in dimethylsulphoxide (DMSO) in the knees once daily for two weeks and as the control group at the same time DMSO was injected into the knees. All rabbits were killed one week after the last injection. Cartilage tissue and synovium were evaluated with a histological scoring system. RESULTS: Histological evaluation of cartilage tissue by H&E staining revealed a significantly reduced average cartilage tissue destruction score of 1.7 in the resveratrol group versus 2.8 in the control group (p = 0.016). Loss of matrix proteoglycan content in cartilage was also much lower, as determined by safranin O staining. Scores of synovial inflammation didn't show difference between groups (1.3 vs 2.2; p = 0.057). CONCLUSION: A characteristic parameter in arthritis is the progressive loss of articular cartilage. This study suggests that intraarticular injections of resveratrol starting at the onset of disease may protect cartilage against the development of experimentally induced OA. PMID- 15883739 TI - Positive feedback regulation of COX-2 expression by prostaglandin metabolites. AB - OBJECTIVES: Previous studies in cell lines and tissues derived from mice lacking genes encoding cyclooxygenase (COX)-1 or -2 have demonstrated compensatory regulation between the two isoenzymes. To determine whether this compensation was driven by a mechanism that controls prostaglandin (PG) levels, we investigated the effects of PG availability on the regulation of COX and cytosolic phospholipase A2 (cPLA2), an upstream enzyme in the PG pathway. MATERIALS AND METHODS: Mouse lung fibroblast cells were treated with various concentrations of PG metabolites including prostaglandin E2 (PGE2), 15-deoxy-delta(12,14) prostaglandin J2 (15d-PGJ2), 6-keto PGF1alpha and PGF2alpha. Cells were harvested for protein and mRNA analyses; culture supernatant was collected for prostaglandin assays. RESULTS: We observed 8- and 20-fold increase in basal COX-2 protein expression levels when cells were exposed to PGE2 and 15d-PGJ2, respectively. In the presence of IL-1beta, PGE2, 15d-PGJ2, 6-keto PGF1alpha and PGF2alpha each enhanced COX-2 protein expression between 5- to 20-fold. Corresponding with the induction of COX-2 protein expression, the latter three PGs induced PGE2 synthesis in a dose-dependent manner. Only PGE2 and 15d-PGJ2 induced COX-2 mRNA expression, although to a lower extent than protein induction. None of the PG metabolites tested showed significant effects on the level of COX 1 or cPLA2 protein expression, except for PGF2alpha, which increased IL-1beta induced cPLA2 protein expression slightly. CONCLUSION: Our results demonstrate that there is positive feedback regulation of COX-2 expression by PG metabolites, but not COX-1, indicating that PG levels per se do not play an important role in the compensatory regulation between the two COX isoenzymes, but may play an important role in mediating increased COX-2 expression and activity. PMID- 15883740 TI - Influence of insulin on the microvascular response to inflammatory mediators in neonatal streptozotocin diabetic rats. AB - OBJECTIVES: To investigate the effect of insulin on microvascular responses to inflammatory mediators in a model of type 2 diabetes mellitus. MATERIALS: We used the neonatal streptozotocin (n-STZ)-induced diabetes model. Diabetes was induced in male newborn (2-day-old) Wistar rats through STZ administration. Experiments were performed 10-12 weeks later. METHODS: Rats were divided into control (sham injected) and study (n-STZ) groups. Using a closed-circuit video camera coupled to a microscope, changes in mesenteric arteriolar and venular diameters induced by topical application of the inflammatory mediators histamine, bradykinin and platelet-activating factor were assessed in chloral hydrate-anesthetized rats. TREATMENT: The n-STZ rats received NPH insulin s.c. for either 4 h or 12 days. RESULTS: Impaired arteriole and venule responses to the inflammatory mediators tested were observed in n-STZ rats. Both acute and chronic insulin treatment corrected the alterations. CONCLUSION: We conclude that insulin is beneficial, restoring microvascular reactivity to inflammatory mediators in type 2 diabetes. PMID- 15883741 TI - Granulocyte colony-stimulating factor therapy and systemic inflammation in critically ill patients. AB - OBJECTIVE AND DESIGN: The effect of the granulocyte colony-stimulating factor filgrastim on systemic inflammation was investigated in a prospective, randomized, placebo-controlled, double-blind study in critically ill patients. SUBJECTS: 59 critically ill patients were recruited within 48 h of intubation due to ventilatory insufficiency. TREATMENT: Subcutaneous dosage of placebo or 300 microg filgrastim once daily. METHODS: Serum samples were collected at study entry, and 1 and 3 days after the start (Day1 and Day3, respectively). Levels of soluble E-selectin (sE-selectin) and interleukin (IL)-10 were determined by ELISA, and those of IL-6, and soluble IL-2 receptor (sIL-2R) by Immulite chemiluminescence immunoassay. RESULTS: The median sE-selectin level decreased by day 3 significantly in the control group but not in the filgrastim group. The difference in the change between the study groups was significant (p = 0.049). IL 10 levels decreased significantly in the filgrastim group, tended to decrease in controls (p = 0.052), and the difference in the change tended to be significant (p = 0.058). IL-6 levels decreased in both groups comparably. sIL-2R levels were elevated and stable. CONCLUSIONS: Filgrastim prolongs endothelial activation and possibly inhibits development of immune suppression mediated by IL-10. PMID- 15883742 TI - The role of protein kinase C in regulating equine eosinophil adherence and superoxide production. AB - OBJECTIVE: To determine if protein kinase C (PKC) regulates equine eosinophil function. MATERIAL OR SUBJECTS: Blood eosinophils were obtained from healthy ponies. METHODS: IL-5- and histamine-induced adherence to serum-coated plastic was measured as the eosinophil peroxidase content of adherent cells and serum treated zymosan (STZ)-and IL-5-induced superoxide production by the reduction of cytochrome C. Eosinophil PKC activity was quantitated as the rate of transfer of (32)P from ATP to substrate. The effects of Ro31-8220 (isotype non-selective PKC inhibitor), Go6976 (conventional PKC inhibitor), and rottlerin (PKCdelta inhibitor) were determined by ANOVA and Bonferroni's or Dunnett's test. RESULTS: Ro31-8220 and Go6976 reduced superoxide production whereas only Go6976 inhibited adherence. Rottlerin inhibited histamine-induced adherence and increased STZ induced superoxide production. Ro31-8220 and Go6976, but not rottlerin, inhibited PKC activity. CONCLUSIONS: PKC is involved in regulating equine eosinophil adherence and superoxide production. The role of PKCdelta appears to depend upon the stimulus used and response measured. PMID- 15883743 TI - Insight into the involvement of Kupffer cell-derived mediators in the hepatoprotective effect of glycine upon inflammation: study on rat precision-cut liver slices. AB - OBJECTIVE AND DESIGN: To investigate the role of inflammatory mediators in the hepatoprotective effect of glycine against lipopolysaccharide (LPS)-induced liver injury in rats. MATERIAL OR SUBJECTS: Male Wistar rats were used (N = 4 or 5 per group). Precision-cut liver slices (PCLS) were prepared for in vitro studies. TREATMENT: Glycine (10 mM) and LPS (10 mug/ml) were added to the incubation medium of PCLS obtained 3 h after LPS intraperitoneal (i. p.) administration (10 mg/kg) or saline injection to rats. Glycine effects were also investigated in vivo by treating rats with a diet containing glycine (5%) during 3 days. METHODS: Tissue injury was assessed by measuring adenosine triphosphate (ATP) and glycogen contents of liver tissue as well as by measuring aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) activity in the medium (in vitro) or in the serum (in vivo). Tumor necrosis factor-alpha (TNF alpha), prostaglandin E(2)(PGE(2)) and NOx (reflecting nitric oxide production) were measured in the incubation medium or in the serum. Histological detection of both ED-2 and peroxidase activity were used as Kupffer cell markers. Student t test or two-way ANOVA were used for statistic analysis. RESULTS: Glycine added to the culture medium increased both ATP and glycogen contents of PCLS from LPS treated rats, reduced the production of TNF-alpha and NOx whereas PGE(2) secretion by PCLS increased. In contrast to the in vitro effect of glycine, we observed that a glycine-enriched diet decreased PGE(2) secretion in the serum after LPS challenge. CONCLUSION: The effect of glycine on LPS-induced mediator secretion is different considering in vitro or in vivo situations. Interestingly, glycine in vitro is able to prevent energy status depletion of PCLS occurring upon inflammation, a phenomenon probably linked to change in inflammatory mediator secretion pattern by hepatic immune cells, namely Kupffer cells. PMID- 15883744 TI - Retinoic acid inhibits CD40 plus IL-4 mediated IgE production through alterations of sCD23, sCD54 and IL-6 production. AB - BACKGROUND: All-trans retinoic acid (ATRA) inhibits IgE synthesis from anti-CD40 plus IL-4 stimulated human B lymphocytes. OBJECTIVE: To study the underlying mechanisms, we examined here molecules which are known to have an impact on IgE production, namely CD23, CD54 and IL-6. METHODS: Human anti-CD40 plus IL-4 stimulated B cells were cultured in the absence and presence of ATRA (10(-6)-10( 10) M). ELISAs were performed to determine soluble (s) CD23 and sCD54, IL-6 and IgE-levels. CD23 and CD54 surface expression were determined by flow cytometric analysis. Semiquantitative-RT-PCR was employed to analyse IL-6, CD23 and CD54 mRNA expression. RESULTS: ATRA induced a dose-dependent increase of percent CD23 (3.4 fold) or CD54 (1.6 fold) positive B cells. At the mRNA level, this was reflected by a modest increase of CD54 mRNA (46.5 +/- 15.8%) only. By contrast, levels of sCD54 were decreased dose-dependently in the presence of ATRA (56.6 +/- 7.6%). Cytokine analysis showed that IL-6 secretion was significantly inhibited by ATRA (53.6 +/- 0.6%) and also IL-6 mRNA synthesis was reduced (66.3 +/- 11.6%). The observed inhibition of IgE production mediated by ATRA was significantly reversed to 90.5 +/- 12% by the addition of 100 pg/mL recombinant IL-6. CONCLUSIONS: ATRA interferes through several pathways with the anti-CD40 plus IL-4 mediated B cell activation, namely IL-6, CD23 and CD54. PMID- 15883745 TI - Pharmacological characterization of P2X7 receptors in rat peritoneal cells. AB - OBJECTIVE AND DESIGN: P2X(7) receptor activation by ATP results in the release of IL-1beta and IL-18. Prolonged stimulation can lead to pore formation and cell death. In this study we pharmacologically characterized P2X(7) receptors on rat peritoneal cells (RPC) and on 1321N1 cells transfected with rat P2X(7) receptor (1321rP2X(7)-11). MATERIALS AND METHODS: RPC were isolated from rats by lavage. P2X(7) agonist induced pore formation in RPC was measured by EtBr uptake. P2X(7) stimulated pore formation and Ca(++) influx in 1321rP2X(7)-11 cells were measured by a fluorometric imaging plate reader. The effects of pyridoxal phosphate-6-azo phenyl -2'-4'-disulfonic acid (PPADS) on pore formation and Ca(++) influx were examined in both RPC and 1321rP2X(7)-11. P2X(7)-mediated IL-1beta release in RPC and the effect of PPADS were determined. RESULTS: RPC express functional P2X(7) receptors that were activated by ATP analogs with a rank order of potency of 2'- 3'-O-(4-Benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) > ATP > alpha,beta methylene ATP. Activation of P2X(7) receptors by BzATP was inhibited by PPADS. Similar results were also obtained in 1321rP2X(7)-11 cells. Activation of P2X(7) receptors on RPC resulted in IL-1 beta secretion, which was inhibited by PPADS. CONCLUSIONS: RPC express functional P2X(7) receptors that form pores and mediate the release of IL-1beta. PMID- 15883746 TI - Preventive actions of a high dose of glucosamine on adjuvant arthritis in rats. AB - OBJECTIVE: Glucosamine, a naturally occurring amino monosaccharide has been used to treat or prevent osteoarthritis in humans. In this study, we evaluated the effect of glucosamine on rat adjuvant arthritis, a model of rheumatoid arthritis. MATERIALS AND METHODS: Adjuvant arthritis was induced in male Wistar rats by injection of Freund's complete adjuvant (FCA) into the right hind paw, and 300 mg/kg of glucosamine, an extra-dose compared with a regular dose for osteoarthritis patients (1.5 g/day, approximately 25 mg/kg), was orally administered once a day to the arthritic rats for 22 days. RESULTS: Glucosamine significantly suppressed the increase in arthritis score (p < 0.05) after day 10 of adjuvant injection, and inhibited the swelling of FCA-injected right and uninjected left hind paws (p < 0.01) after day 18. In addition, histopathological examination of the arthritic joints revealed that glucosamine suppressed synovial hyperplasia, cartilage destruction and inflammatory cell infiltration. Furthermore, glucosamine reduced the production of nitric oxide and prostaglandin E(2) in plasma (p < 0.05). CONCLUSIONS: These observations suggest that glucosamine is able to suppress the progression of adjuvant arthritis in rats. Glucosamine may be expected as a novel anti-inflammatory agent for treatment of rheumatoid arthritis. PMID- 15883747 TI - Large dose ketamine inhibits lipopolysaccharide-induced acute lung injury in rats. AB - BACKGROUND: Sepsis is associated with the highest risk of progression to acute lung injury or the acute respiratory distress syndrome. Ketamine has been advocated for anesthesia in endotoxemic and other severely ill patients because it is a cardiovascular stimulant. Our study was designed to investigate the effect of ketamine on the endotoxin-induced acute lung injury in vivo. MATERIALS AND METHODS: Adult male Wistar rats were randomly divided into 6 groups: saline controls; rats challenged with endotoxin (5 mg/kg) and treated with saline; challenged with endotoxin (5 mg/kg) and treated with ketamine (0.5 mg/kg); challenged with endotoxin (5 mg/kg) and treated with ketamine (5 mg/kg); challenged with endotoxin (5 mg/kg) and treated with ketamine (50 mg/kg); saline injected and treated with ketamine (50 mg/kg). TNF-alpha, IL-6 and NF-kappa B were investigated in the tissues of the lung after 2 h. Myeloperoxidase (MPO) activity and wet/dry weight ratio were investigated 6 h later. RESULTS: We demonstrated that intravenous administration of endotoxin could provoke significant lung injury, which was characterized by increase of MPO activity and wet/dry weight ratio, TNF-alpha and IL-6 expression and NF-kappa B activation. Ketamine (5, 50 mg/kg) inhibited endotoxin-induced NF-kappa B activation. Ketamine only at a dose of 50 mg/kg inhibited TNF-alpha and IL-6 production, and decreased MPO activity and wet/dry weight ratio after endotoxin challenge. CONCLUSIONS: Ketamine, only at a supra-anesthetic dosage, could inhibit endotoxin induced pulmonary inflammation in vivo. PMID- 15883748 TI - The influence of vitamin C on systemic markers of endothelial and inflammatory cell activation in smokers and non-smokers. AB - OBJECTIVE AND DESIGN: To determine the influence of vitamin C supplementation (500 mg, bd, 14 days) on the circulating concentrations of soluble ICAM-1 (a marker of endothelial activation), neopterin (a marker of monocyte activation), and neutrophil elastase (a marker of neutrophil activation) in smokers and non smokers in a randomised, double-blind, placebo-controlled trial in a hospital setting. SUBJECTS: Twenty smokers (serum cotinine > or = 20 ng ml(-1)) and 20 age and gender-matched non-smokers (serum cotinine < or = 13.7 ng ml(-1)). RESULTS: At baseline, there was a significant elevation in the concentration of sICAM-1 in smokers (median 247, IQR 199 to 357 ng ml(-1)) compared to non-smokers (median 207, IQR 189 to 227 ng ml(-1); p = 0.014). Vitamin C supplementation did not influence the circulating concentrations of ICAM-1 or neopterin, or leukocyte elastase activity, in smokers, non-smokers, or in the total population. CONCLUSIONS: Markers of monocyte and neutrophil activation were not influenced by smoking status in this study population. However, sICAM-1 concentrations were significantly elevated in tobacco smokers, reflecting tobacco-induced vascular activation that is unaffected by Vitamin C supplementation. PMID- 15883749 TI - Clonidine--revisited, relived or time for further investigations? PMID- 15883750 TI - Erythropoietin: from hematopoiesis to cardioprotection. PMID- 15883751 TI - Glucagon like peptide-1 is protective against myocardial ischemia/reperfusion injury when given either as a preconditioning mimetic or at reperfusion in an isolated rat heart model. PMID- 15883752 TI - Simvastatin regulates myocardial cytokine expression and improves ventricular remodeling in rats after acute myocardial infarction. AB - PURPOSE: Studies have showed that inflammatory cytokines were involved in the process of left ventricular (LV) remodeling after acute myocardial infarction (AMI), anti-inflammation treatment ameliorated LV remodeling and improved cardiac performance. Hydroxymethylglutary coenzyme A reductase inhibition (statins) could affect the expression of inflammatory cytokines. We hypothesized that statins have beneficial effects on early LV remodeling and cardiac performance in rats with AMI by modulating the production of inflammatory cytokines. METHODS: Rats with AMI were treated with placebo or simvastatin (gastric gavage) for 4 weeks. The pro-inflammatory cytokines: tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6 and the anti-inflammatory cytokine: IL-10 excreted by cardiac myocytes was examined. Echocardiography, hemodynamics and collagen type I production were measured to evaluate LV remodeling and cardiac function. RESULTS: The mRNA expression and protein production of TNF-alpha, IL-1beta, IL-6 and IL-10 in AMI group were significantly elevated compared with sham rats. Simvastatin markedly attenuated the production of TNF-alpha, IL-1beta, IL-6 and increased IL 10 levels in the noninfarcted and infarcted regions, reduced collagen deposition in the noninfarcted myocardium and improved left ventricular function. However simvastatin did not alter plasma lipids. CONCLUSIONS: Simvastatin ameliorates early LV remodeling and improve cardiac function after AMI. Simultaneously, it decreased pro-inflammatory and increased anti-inflammatory cytokines, which suggests, but does not prove, a causal relationship independent of plasma lipid lowering effects. PMID- 15883753 TI - Effect of sildenafil on reperfusion function, infarct size, and cyclic nucleotide levels in the isolated rat heart model. AB - We have previously shown that NO-donor induced elevation in myocardial cGMP levels is associated with improved reperfusion function of the isolated rat heart. The phosphodiesterase 5 (PDE 5) inhibitor, sildenafil could potentially increase myocardial cGMP levels and thus protect the heart against ischaemic/reperfusion injury. METHODS: To test our hypothesis we treated the isolated working rat heart with vehicle, OR sildenafil (10, 20, 50, 100, 200 nM), OR sildenafil (50 nM) plus a sarcolemmal (HMR 1098) or a mitochondrial (5 Hydroxydecanoate (5-HD)) K(ATP) channel blocker. Hearts were then subjected to 20 min global, or 35 min regional ischaemia at 37( composite function)C before reperfusion function (aortic output, coronary flow and aortic pressure) and infarct size were documented. Pre-ischaemic, ischaemic and reperfusion myocardial cAMP and cGMP concentrations were determined. RESULTS: Low concentrations of sildenafil (10, 20 and 50 nM) improved reperfusion aortic output (AO) recovery (61.4+/- 4.5%, 64.8 +/- 5.2% and 62.3 +/- 5.0% vs. 45.4 +/- 3.8% for controls (p < 0.05)) and infarct size, while high concentrations (200 nM) worsened AO recovery (24.9 +/- 4.9.0%, p < 0.05). Myocardial cGMP levels of ischaemic tissue were elevated (34.7 +/- 2.4 vs. 27.3 +/- 2.2 pmol/g ww) and cAMP levels were suppressed (0.59 +/- 0.03 vs. 0.87 +/- 0.06 nmol/g ww) in the sildenafil (50 nM) treated hearts. Co-perfusion with sildenafil plus HMR 1098 decreased AO recovery (21.7 +/- 7.6% vs. 62.3 +/- 5.0% for sildenafil alone, p < 0.05) and increased infarct size (29.7 +/- 2.04% vs. 8.6 +/- 2.39% for sildenafil alone, p < 0.05).Similarly, sildenafil plus 5-HD decreased reperfusion AO recovery (44.4 +/- 6.0% vs. 62.3 +/- 5.0% for sildenafil alone, p < 0.05) and increased infarct size (33.8 +/- 1.62% vs. 8.6 +/- 2.39% for sildenafil alone, p < 0.05). CONCLUSIONS: (1) Pretreatment with low concentrations of sildenafil (20-50 nM) improves, while higher concentrations (200 nM) worsen reperfusion function in this model. (2) Low concentrations of sildenafil (20-50 nM) decrease infarct size while the higher concentrations had no effect. (3) These protective properties of low concentrations of sildenafil may be related to its cGMP elevating and cAMP suppressing effects in the ischaemic heart. (4) Possible end-effectors for sildenafil in the ischaemic heart include the mitochondrial and sarcolemmal K(ATP) channel. PMID- 15883754 TI - Erythropoietin just before reperfusion reduces both lethal arrhythmias and infarct size via the phosphatidylinositol-3 kinase-dependent pathway in canine hearts. AB - Although recent studies suggest that erythropoietin (EPO) may reduce multiple features of the myocardial ischemia/reperfusion injury, the cellular mechanisms and the clinical implications of EPO-induced cardioprotection are still unclear. Thus, in this study, we clarified dose-dependent effects of EPO administered just before reperfusion on infarct size and the incidence of ventricular fibrillation and evaluated the involvement of the phosphatidylinositol-3 (PI3) kinase in the in vivo canine model. The canine left anterior descending coronary artery was occluded for 90 min followed by 6 h of reperfusion. A single intravenous administration of EPO just before reperfusion significantly reduced infarct size (high dose (1,000 IU/kg): 7.7 +/- 1.6%, low dose (100 IU/kg): 22.1 +/- 2.4%, control: 40.0 +/- 3.6%) in a dose-dependent manner. Furthermore, the high, but not low, dose of EPO administered as a single injection significantly reduced the incidence of ventricular fibrillation during reperfusion (high dose: 0%, low dose: 40.0%, control: 50.0%). An intracoronary administration of a PI3 kinase inhibitor, wortmannin, blunted the infarct size-limiting and anti-arrhythmic effects of EPO. Low and high doses of EPO equally induced Akt phosphorylation and decreased the equivalent number of TUNEL-positive cells in the ischemic myocardium of dogs. These effects of EPO were abolished by the treatment with wortmannin. In conclusion, EPO administered just before reperfusion reduced infarct size and the incidence of ventricular fibrillation via the PI3 kinase dependent pathway in canine hearts. EPO administration can be a realistic strategy for the treatment of acute myocardial infarction. PMID- 15883755 TI - Antiplatelet and antithrombotic activity of cilostazol is potentiated by dipyridamole in rabbits and dissociated from bleeding time prolongation. AB - PURPOSE: To determine the antiplatelet effect of cilostazol (Pletal) and its interaction with dipyridamole in in vitro and in vivo rabbit models, and to see if it can be dissociated from bleeding time prolongation. METHODS: In vitro collagen-induced platelet aggregation was measured by an impedance-based aggregometer. The in vivo antithrombotic effect was evaluated in a rabbit carotid artery cyclic flow reduction (CFR) model, in which repetitive thrombosis was induced by mechanical injuries of the artery and stenosis. Template bleeding time was determined in rabbit ear arterioles and hindlimb nail cuticles. RESULTS: In vitro platelet aggregation was slightly inhibited by 4 microM cilostazol (22 +/- 6%), and modestly by 13 microM (57 +/- 3% of aggregation). While dipyridamole itself up to 13 microM had no significant inhibition, it potentiated the effect from cilostazol: in the presence of 4 microM dipyridamole, 4 microM cilostazol inhibited aggregation by 47 +/- 6%. Dipyridamole also potentiated the CFR reducing effect of cilostazol: combination of dipyridamole (no effect by itself) and cilostazol at 1 microM decreased CFRs to levels achieved by 3-4 microM cilostazol alone. Bleeding times were similar in controls and animals treated with cilostazol, or with cilostazol and dipyridamole. In contrast, aspirin (4 mg/kg), while reducing CFRs, significantly increased bleeding time. CONCLUSION: These results suggest that dipyridamole potentiates the antiplatelet effect of cilostazol without prolongation of the bleeding time, implying a potential novel combination antithrombotic therapy. PMID- 15883756 TI - Clonidine lowers blood pressure by reducing vascular resistance and cardiac output in young, healthy males. AB - PURPOSE: Clonidine is a classical sympatholytic drug that is widely used for the treatment of hypertension. Experimental and clinical studies suggest that Clonidine may activate baroreflex. The aim of this study was to determine the hemodynamic response to Clonidine under physiological conditions and to test the hypothesis that Clonidine would reduce cardiac output and blood pressure resulting in an increase in total peripheral resistance. METHODS: Clonidine's hemodynamic effect was evaluated in 28 young, healthy subjects after a single i.v. dose of 1 microg x kg(- 1). Impedance cardiography, systolic time intervals and pulse wave analysis were used to characterize myocardial and vascular function. RESULTS: Clonidine lowered blood pressure, heart rate, left ventricular ejection time, and pulse wave velocity and increased pre-ejection period. Stroke volume and cardiac output decreased gradually over the investigation time of 240 min. Central systolic blood pressure (SBP) was lowered to a larger extent than peripheral SBP. Total peripheral resistance was characterized by an immediate fall of short duration followed by a continuous rise above baseline after 120 min. Placebo did not have any significant effect on hemodynamic parameters. CONCLUSIONS: Clonidine's blood pressure lowering effect is mediated by both an immediate decrease in vascular resistance and a prolonged decrease in cardiac output, and Clonidine lowers central SBP more than peripheral SBP. PMID- 15883757 TI - Clinical comparison of ibutilide and propafenone for converting atrial flutter. AB - OBJECTIVE: To evaluate the efficacy and safety of intravenous ibutilide and propafenone for immediate treatment of atrial flutter. METHODS: Forty patients with atrial flutter with an arrhythmia duration of three hours to 90 days were randomized to receive up to two 10-minute infusions of ibutilide (1 and 1 mg) or propafenone (70 and 70 mg) with a 10-minute interval. RESULTS: Ibutilide was superior to propafenone for treating atrial flutter (90% vs. 30%, p < 0.01). The median conversion time in the ibutilide group was 11 min (the 25th and 75th percentile was 10 and 45 min), and the median conversion time in the propafenone group was 35 min (range 20-55 min). In all patients, the duration of arrhythmia before treatment was a predictor of arrhythmia termination, although this was more obvious in the group that received ibutilide. Conversion of atrial flutter by ibutilide was characterized mainly by increased cycle length variability. Bradycardia (2/20) and hypotension (4/20) were more common side effects with propafenone. Of 20 patients given ibutilide, 8 (40%) who developed monomorphic ventricular extrasystoles or repetitive atrial flutter with aberrant conduction tachycardia, no one required any specific treatment except for the interruption of ibutilide infusion. CONCLUSION: Ibutilide is highly effective for rapidly terminating atrial flutter. This new class III drug, under monitored conditions, is a potential alternative to currently available cardioversion options. PMID- 15883758 TI - Urotensin II: its function in health and its role in disease. AB - Urotensin II (U-II) is the most potent vasoconstrictor known, even more potent than endothelin-1. It was first isolated from the fish spinal cord and has been recognized as a hormone in the neurosecretory system of teleost fish for over 30 years. After the identification of U-II in humans and the orphan human G-protein coupled receptor 14 as the urotensin II receptor, UT, many studies have shown that U-II may play an important role in cardiovascular regulation. Human urotensin II (hU-II) is an 11 amino acid cyclic peptide, generated by proteolytic cleavage from a precursor prohormone. It is expressed in the central nervous system as well as other tissues, such as kidney, spleen, small intestine, thymus, prostate, pituitary, and adrenal gland and circulates in human plasma. The plasma U-II level is elevated in renal failure, congestive heart failure, diabetes mellitus, systemic hypertension and portal hypertension caused by liver cirrhosis. The effect of U-II on the vascular system is variable, depending on species, vascular bed and calibre of the vessel. The net effect on vascular tone is a balance between endothelium-independent vasoconstriction and endothelium dependent vasodilatation. U-II is also a neuropeptide and may play a role in tumour development. The development of UT receptor antagonists may provide a useful research tool as well as a novel treatment for cardiorenal diseases. PMID- 15883759 TI - Rethinking the renin-angiotensin system and its role in cardiovascular regulation. AB - Angiotensin-converting enzyme (ACE) plays a pivotal role in the renin-angiotensin system (RAS) and ACE-inhibitors are widely used in several clinical conditions, including hypertension and heart failure. Recently, a homologue of ACE, ACE(2) has been discovered. Both ACE and ACE(2) are emerging as key enzymes of the RAS, where ACE(2) may play a role as negative regulator of ACE. Moreover, ACE(2) appears to be an important enzyme outside the classical RAS, as it hydrolyzes apelins, dynorphin A 1-13, des-Arg-bradykinin and other peptide substrates. The precise interplay between tissue ACE, ACE(2), and their substrates and by products are presently still unclear.ACE-inhibitors reduce angiotensin II formation and bradykinin degradation, but do not inhibit ACE(2) activity. Moreover, ACE-inhibitors differ in their affinity for tissue ACE, and it has been suggested that tissue ACE affinity might be responsible for some of the beneficial properties of these drugs. ACE-inhibitors also increase nitric oxide availability, and activate several kinases that may regulate protein synthesis by interacting with the nucleus of the cells (outside-in signaling). The outside-in signaling may also be activated by bradykinin itself. Although, the precise significance of the outside-in signaling is still unclear, this new role of ACE inhibitors may represent a discriminant factor versus angiotensin II receptors antagonists. This mini review will summarize some new aspects concerning the recently discovered biological functions of RAS and in particular of ACE, ACE(2) and ACE-inhibitors in cardiovascular system. PMID- 15883760 TI - Effect of a histone deacetylase inhibitor on human cardiac mass. PMID- 15883762 TI - Metal-enhanced fluorescence from plastic substrates. AB - We report the first findings of Metal-Enhanced Fluorescence (MEF) from modified plastic substrates. In the past several years our laboratories have reported the favorable effects of fluorophores in close proximity to silver nanoparticles. These effects include, enhanced fluorescence intensities, (increased detectability), and reduced lifetimes, (enhanced fluorophore photostability). All of these reports have featured silver nanostructures and fluorophores which have been immobilized onto clean glass or quartz surfaces. In this report we show how plastic surfaces can be modified to obtain surface functionality, which in turn allows for silver deposition and therefore metal-enhanced fluorescence of fluorophores positioned above the silver using a protein spacer. Our findings show that plastic substrates are ideal surfaces for metal-enhanced phenomena, producing similar enhancements as compared to clean glass surfaces. Subsequently, we speculate that plastic substrates for MEF will find common place, as compared to the more expensive and less versatile traditional silica based supports. PMID- 15883763 TI - Fluorescent characteristics of rheumatoid arthritis patients blood lymphocytes. AB - A fluorescent probe, ABM, aminoderivative of benzanthrone, synthesized in the Department of Organic Chemistry of the Riga Technical University (Latvia), has been successfully used to characterize changes in the structural and functional properties of cell membranes during different pathologies. In the present study the physicochemical properties and the functional activity of the peripheral blood mononuclear cells (lymphocytes-Ly) in patients with rheumatoid arthritis (RA) were studied using the ABM probe. Intensity of the ABM fluorescence in the celi suspension, functional activity of the ly anisotropy of the membranes differ patients with different titres of rheumatoid factor in blood. Patients with seropositive RA had decreased proliferative activity and lower number of iy in blood plasma indicating greater alterations of the immunoregulating processes in these patients as compared to patients with seronegative RA. In the latter the Ly deficiency is compensated to some extent by increased proliferation activity of these cells. The ABM fluorescence intensity correlated not only with membrane anisotropy (r = 0.97, but also with the proliferation activity of the Ly (r = 0.98). The above parameters correlated with the clinical manifestations of the disease. The results indicate that the fluorescent probe ABM is useful for screening the physicochemical status of Ly membranes and the proliferation activity of these cells in RA patients. PMID- 15883764 TI - The dynamics of intramolecular excited state relaxation of N-anthryl substituted pyridinium cations. AB - N-(1-Anthryl)-2,4,6-trimethyl-pyridinium (I), N-(2-anthryl)-2,4,6-trimethyl pyridinium (II) and 10-(1-anthryl)-1,2,3,4,5,6,7,8-octahydro-acridinium cations (III) with anomalously high fluorescence Stokes' shift have been investigated. Fluorescence kinetics analysis at various temperatures showed that in the range 293-77 K, the radiative deactivation rate constants (kf) increase by 5.5 to 30 times. The low-temperature time-resolved emission spectra of I-III were found to be consistent with the model: A--> A*<--> B* where A* is the local excited twisted form and B* is the relaxed more planar, bent conformer of the molecule. The rate constants of the excited relaxed state formation (k1) and back reaction (k-1) of compounds studied were estimated. PMID- 15883765 TI - Fluorimetry studies of oscillating chemilumnescence in the luminol-H2O2-KSCN CuSO4-TMAOH system. AB - Oscillating chemical reactions are complex systems, involving a large number of chemical species. In oscillating chemical reaction, some species, usually a reaction intermediate, exhibit fluctuation in its concentration. In this report, visible oscillating chemiluminescence produced by the addition of luminol (3 aminophthalhydrazide) to the oscillating system of H2O2-KSCN-CuSO4-TMAOH was investigated using spectrofluorimetry method. The effects of ingredient concentration of the oscillating system and complexing agents like citric acid and cysteine on the behavior of the oscillating system were investigated. Moreover, the influence of nonaqueous solvents such as ethanol and ethylene glycol has been studied. PMID- 15883766 TI - Analysis of chlorophyll fluorescence spectra in some tropical plants. AB - Laser-induced chlorophyll fluorescence (LICF) emission spectra of leaves of some tropical plants were measured using a compact fiber-optic fluorosensor with a continuous-wave blue diode laser as exciting source and an integrated digital spectrometer. Different chlorophyll-fluorescence signatures of light-green, fully green, and yellow leaves were monitored at room temperature. Deconvolution procedure was used to determine fluorescence band position and width. Calibration of the fluorescence ratio F690/F730 relative to the 404 nm excitation is done from the curve-fitted parameter. PMID- 15883767 TI - A sensitive fluorimetric method for the determination of epinephrine. AB - A sensitive fluorimetric method for the determination of epinephrine (E) is described in this paper. The experiments indicate that epinephrine can react with formaldehyde (HCHO) in an acid medium to form a condensation product, which can be oxidized by potassium hexacyanoferrate(III) (K3[Fe(CN)6]) in borax buffer (pH = 9.5). The reaction product can emit strong fluorescence. Ascorbic acid (AA) is used in order to consume excess potassium hexacyanoferrate and stabilize the fluorescent product. Under optimum conditions, a linear relationship has been obtained between the fluorescence intensity and the concentration of epinephrine in the range of 1.4 x 10(-9) - 2.1 x 10(-6) mol/l, and the detection limit is 2.4 x 10(-10) mol/l (4.3 x 10(-11) g/ml, S/N = 3). The method is applied for the determination of E in both actual sample and the synthetic sample with E and norepinephrine (NE) by using the coupling technique of synchronous fluorimetry and H-point standard addition method, and the results obtained are satisfactory. PMID- 15883768 TI - Hemolysis of erythrocytes and erythrocyte membrane fluidity changes by new lysosomotropic compounds. AB - This work contains the results of studies on the influence of newly synthesized lysosomotropic substances (lysosomotropes) on human erythrocytes. Six homologous series of the compounds differing in the alkyl chain length and counterions were studied. They were found to hemolyse erythrocytes and to change their osmotic resistance. The observed hemolytic effects were dependent both on the compound's structure (polar head dimension and alkyl chain length of compound) and its form (the kind of the counterion). In parallel, the influence of lysosomotropes on fluidity of the erythrocyte membrane was studied. Three different fluorescent probes were used; 1,6-diphenyl-1,3,5-hexatriene (DPH), 1-(4 trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene, p-toluenesulfonate (TMA-DPH) and 6-dodecanoyl-2-dimethylaminonaphthalene (laurdan). Their anisotropy (DPH and TMA-DPH) or general polarization (laurdan) values after incorporation into ghost erythrocyte membranes were measured. The results obtained show that fluidity changes accompanied the effects observed in hemolytic experiments both quantitatively and qualitatively. PMID- 15883769 TI - Fluorescent methods to study DNA, RNA, proteins and cytoplasmic membrane polarization in the pentachlorophenol-mineralizing bacterium Sphingomonas sp. UG30 during nutrient starvation in water. AB - The effect of sodium pentachlorophenolate (NaPCP) exposure on the nutrient starved pentachlorophenol (PCP)-mineralizing bacterium Sphingomonas sp. UG30 was assessed using fluorescent methods to measure DNA, RNA, total cellular protein, and cytoplasmic membrane proteins. UG30 cells were inoculated into sterilized Speed River (Guelph, ON, Canada) water samples in the presence of 50, 100, and 250 ppm NaPCP. No marked changes were observed in the total cellular DNA, RNA or protein levels over 90 d, indicating the macromolecular composition of UG30 was not affected by both nutrient limitation and NaPCP. Total cell counts as determined by DAPI staining also did not change over 90 d. Over the same period, viable counts decreased with increasing concentrations of NaPCP. At 250 ppm NaPCP, viable cell counts decreased over 6 orders of magnitude after 1 hr exposure. Cell numbers partially recovered once NaPCP was degraded. The UG30 cytoplasmic membrane polarization ratio also decreased after NaPCP was depleted. The decreased polarization value at the end of the study period suggested the UG30 membrane was more fluid and that this increase in fluidity was due to nutrient starvation effects rather than exposure to NaPCP. These results indicated that UG30 is a robust organism that is able to degrade NaPCP even under adverse conditions and fluorescent methods are useful for determining macromolecular concentrations and cytoplasmic membrane polarization values. PMID- 15883770 TI - Effects of refractive index and viscosity on fluorescence and anisotropy decays of enhanced cyan and yellow fluorescent proteins. AB - The fluorescence lifetime strongly depends on the immediate environment of the fluorophore. Time-resolved fluorescence measurements of the enhanced forms of ECFP and EYFP in water-glycerol mixtures were performed to quantify the effects of the refractive index and viscosity on the fluorescence lifetimes of these proteins. The experimental data show for ECFP and EYFP two fluorescence lifetime components: one short lifetime of about 1 ns and a longer lifetime of about 3.7 ns of ECFP and for EYFP 3.4. The fluorescence of ECFP is very heterogeneous, which can be explained by the presence of two populations: a conformation (67% present) where the fluorophore is less quenched than in the other conformation (33% present). The fluorescence decay of EYFP is much more homogeneous and the amplitude of the short fluorescence lifetime is about 5%. The fluorescence anisotropy decays show that the rotational correlation time of both proteins scales with increasing viscosity of the solvent similarly as shown earlier for GFP. The rotational correlation times are identical for ECFP and EYFP, which can be expected since both proteins have the same shape and size. The only difference observed is the slightly lower initial anisotropy for ECFP as compared to the one of EYFP. PMID- 15883771 TI - Optimization of gold nanoparticle-based DNA detection for microarrays. AB - DNA microarrays are promising tools for fast and highly parallel DNA detection by means of fluorescence or gold nanoparticle labeling. However, substrate modification with silanes (as a prerequisite for capture DNA binding) often leads to inhomogeneous surfaces and/or nonspecific binding of the labeled DNA. We examined both different substrate cleaning and activating protocols and also different blocking strategies for optimizing the procedures, especially those for nanoparticle labeling. Contact angle measurements as well as fluorescence microscopy, atomic force microscopy (AFM), and a flatbed scanner were used to analyze the multiple-step process. Although the examined different cleaning and activating protocols resulted in considerably different contact angles, meaning different substrate wettability, silanization led to similar hydrophobic surfaces which could be revealed as smooth surfaces of about 2-4 nm roughness. The two examined silanes (3-glycidoxypropyltrimethoxysilane (GOPS) and 3 aminopropyltriethoxysilane (APTES)) differed in their DNA binding homogeneity, maximum signal intensities, and sensitivity. Nonspecific gold binding on APTES/PDC surfaces could be blocked by treatment in 3% bovine serum albumin (BSA). PMID- 15883772 TI - General and specific solvent effects in optical spectra of ortho-aminobenzoic acid. AB - We describe studies about solvent effects on the absorption and emission properties of o-aminobenzoic acid (o-Abz), interpreting the results within the framework of general and specific solute-solvent interactions. Measurements were performed in several solvents and analysis of the absorption and emission wavelengths were made based on Lippert's model for general solvent effects and on the use of different parameters to describe the ability of the solvent to promote specific interactions with the solute. We observed low sensitivity of the Stokes shift upon changes in the medium polarity, and large deviation from the linearity predicted by Lippert's equation when the solvents were characterized as Bronsted acid in the Kamlet-Taft pi* scale. Quantum yield and fluorescence lifetimes were best interpreted based on the AN+DN scale used to describe the electron donor/acceptor properties of the solvent. The results indicated that non radiative deexcitation processes are favoured in solvents which promote the formation of intramolecular hydrogen bond, while interactions with electron acceptor solvents lead to enhancement of fluorescence. PMID- 15883773 TI - Propidium iodide and PicoGreen as dyes for the DNA fluorescence correlation spectroscopy measurements. AB - Many experimental designs, in which nucleic acid conformational changes are of interest, require reliable fluorescence labeling. The appropriate fluorescence probe should have suitable optical properties and, more importantly, should not interfere with the investigated processes. In order to avoid chemical modifications the fluorescence label needs to be associated with nucleic acid via weak non-covalent interactions. There are a number of fluorescent probes that change their fluorescent properties (i.e. their quantum yield and/or spectral characteristics) upon association with nucleic acid. Such probes are frequently used to detect, visualize and follow processes involving nucleic acid and its conformational changes. In order to obtain reliable data regarding macromolecule or aggregate topology a detailed knowledge of probe-nucleic acid interactions on the molecular level is needed. In this paper we show that the association of propidium iodide with DNA alters its conformation and that it selectively labels plasmid fragments and/or its subpopulations in a concentration-dependent meaner. Another dye, PicoGreen, exhibits better properties. It labels nucleic acid uniformly and without any concentration-dependent artifacts. PMID- 15883774 TI - Fluorescence of metal-ligand complexes of mono- and di-substituted naphthalene derivatives. AB - In this work, metal ion complexes for several naphthalene derivatives have been investigated. Different working pH values were chosen: 2.5 for complexes with Zr(IV), 4.0 for complexes with Fe(III), 5.0 for complexes with Al(III), and 7.5 for complexes with Cu(II). A stoichiometry of 1:1 for all complexes except two has been established by use of the Benesi-Hildebrand method and the stability constants have been calculated. All complexes between naphthalene derivatives and Cu(II) and Fe(III) show fluorescence quenching. In the case of Al(III), all complexes provided enhanced fluorescence. For Zr(IV), only the complex with 3 hydroxy-2-naphthoic acid provided enhanced fluorescence. The value of the stability constants as a function of the substituents of naphthalene derivatives has been analyzed. One can conclude that Cu(II) showed the largest binding affinity for the mono-substituted derivatives. However, Al(III) and Zr(IV) produced greater selectivity for the di-substituted derivatives. Iron(III) showed no specific binding with any of the naphthalene derivatives. PMID- 15883781 TI - Isethionate as a product from taurine during nitrogen-limited growth of Klebsiella oxytoca TauN1. AB - Klebsiella oxytoca TauN1 represents a group of isolates which utilise taurine (2 aminoethanesulfonate) quantitatively as a sole source of combined nitrogen for aerobic growth. During growth, a compound is excreted, which has now been identified as isethionate (2-hydroxyethanesulfonate). An ion-chromatographic separation of isethionate was developed to quantify the putative isethionate, whose identity was confirmed by matrix-assisted, laser-desorption ionisation time of-flight mass spectrometry. Strain TauN1 utilised taurine (and excreted isethionate) concomitantly with growth. Cell-free extracts contained inducible taurine transaminase, which yielded sulfoacetaldehyde. A soluble, NADP-dependent isethionate dehydrogenase converted sulfoacetaldehyde to isethionate. The enzyme was partially purified and it apparently belonged to the family of short-chain alcohol dehydrogenases. PMID- 15883782 TI - Purification and characterization of succinate:menaquinone oxidoreductase from Corynebacterium glutamicum. AB - Succinate:menaquinone oxidoreductase from Corynebacterium glutamicum, a high-G+C, Gram-positive bacterium, was purified to homogeneity. The enzyme contained two heme B molecules and three polypeptides with apparent molecular masses of 67, 29 and 23 kDa, which corresponded to SdhA (flavoprotein), SdhB (iron-sulfur protein), and SdhC (membrane anchor protein), respectively. In non-denaturating polyacrylamide gel electrophoresis, the enzyme migrated as a single band with an apparent molecular mass of 410 kDa, suggesting that it existed as a trimer. The succinate dehydrogenase activity assayed using 2,3-dimethoxy-5-methyl-6-decyl-1,4 benzoquinone and 2,6-dichloroindophenol as the electron acceptor was inhibited by 2-n-heptyl-4-hydroxyquinoline N-oxide (HQNO), and the Dixon plots were biphasic. In contrast, the succinate dehydrogenase activity assayed using phenazine methosulfate and 2,6-dichloroindophenol was inhibited by p-benzoquinone and not by HQNO. These findings suggested that the C. glutamicum succinate:menaquinone oxidoreductase had two quinone binding sites. In the phylogenetic tree of SdhA, Corynebacterium species do not belong to the high-G+C group, which includes Mycobacterium tuberculosis and Streptomyces coelicolor, but are rather close to the group of low-G+C, Gram-positive bacteria such as Bacillus subtilis. This situation may have arisen due to the horizontal gene transfer. PMID- 15883783 TI - Gene expression profiles of different clinical stages of colorectal carcinoma: toward a molecular genetic understanding of tumor progression. AB - BACKGROUND AND AIMS: Colorectal cancer is one of the leading causes of cancer deaths in the Western world. A better understanding of the development and progression of colorectal carcinoma is needed to define novel targets and strategies for treatment. PATIENTS/METHODS: Gene expression profiles were determined for primary tumors of 10 locally restricted (T3N0M0), 8 lymphatically metastasized (T3N+M0), 7 systemically metastasized (T3N+M1) colorectal carcinomas, and 6 specimens of normal colorectal tissue by histology-guided oligonucleotide microarray analysis. RESULTS: A total of 1,995 genes were differently regulated in primary tumors of colorectal carcinoma compared with normal colorectal tissue. Besides common features of dedifferentiation and different expression of genes involved in cell division, cell adhesion, angiogenesis, signal transduction and metabolism we observed a deregulation of genes with an as yet unclear function. We identified 126 genes that were subsequently up- and 204 genes down-regulated during tumor progression. Furthermore, we found a cluster of five genes exclusively up-regulated in primary tumors of systemically metastasized colorectal carcinomas. A comparison of locally restricted (T3N0M0) and systemically metastasized (T3N+M1) primary tumors showed 50 deregulated genes with a massive down-regulation of immune-modulatory genes in primary tumors of systemically metastasized carcinomas. Primary tumors of lymphatically (T3N+M0) and systemically metastasized (T3N+M1) carcinomas differed in the expression of 19 genes. CONCLUSION: These results provide an additional step toward the identification of crucial genes for the progression of colorectal cancer and the identification of novel treatment targets or strategies. PMID- 15883784 TI - [Plantar fasciitis and radiotherapy. Clinical and radiobiological treatment results]. AB - Patients with plantar fasciitis and pain refractory to conventional therapy are treated with low-dose radiotherapy (RT), but no conclusive evidence-based and radiobiological studies had been performed. In 2001 the German Cooperative Group on Radiotherapy for Benign Diseases (GCG-BD) carried out a study by mailing a standardized questionnaire. A total of 136 institutions treated 3621 patients/year with chronic or refractory pain. The median total dose was 6 Gy (median single dose: 1 Gy); 76 institutions reported data of their clinical evaluation of a total of 7947 patients. Pain relief lasting for at least 3 months was reported in 70% and persistent pain relief in 65%. There were no acute or chronic radiogenic side effects observed. The radiobiological studies showed a significant increase of granulocyte function at 1.5 Gy and a significant decrease at 3.5 and 4.0 Gy. These results may provide a possible explanation for a local anti-inflammatory effect of low-dose RT. RT may be an excellent alternative for patients with contraindications to long-term treatment with steroids or NSAID. PMID- 15883785 TI - [Functional tendon repair in orthopedic tumor surgery]. AB - Large defects of functional soft tissue structures, e.g., extensor mechanism, ligaments, muscles, and joint capsule, may occur in tumor surgery or revision surgery following conventional joint replacement. Reconstruction can be performed using biological grafts such as free or pedicled tendon-muscle flaps, allografts, or synthetic material. Prerequisites for synthetic material are good biologic tolerance with fibroblastic ingrowth, mechanical resistance to fatigue, and a maximum of tension force with a minimum of elongation. In this study we used a nonresorbable band of longitudinal polyester fibers with a minimal rupture level of 4000 N and an elongation rate less than 7% of its original length. The shape of the band was designed for universal use with 40 cm length and 6 cm width. Its primary indication was augmentation or complete reconstruction of the extensor mechanism of the knee joint after large extra-articular tumor resections in primary bone tumors. Furthermore, its use for hip joint capsule reconstruction in luxation, coverage of megaprostheses of the humerus, and augmentation after biological reconstruction of tendons achieved excellent results. PMID- 15883786 TI - Gas sensing using edge-plane pyrolytic-graphite electrodes: electrochemical reduction of chlorine. AB - The voltammetric responses of chlorine in aqueous acid solutions have been explored using different carbon-based electrodes. Edge-plane pyrolytic graphite has more electrochemical reversibility than glassy carbon, basal-plane pyrolytic graphite, or boron-doped diamond electrodes. A significant reduction in the over potential is observed on the edge-plane pyrolytic-graphite electrode in contrast with the other carbon-based electrode substrates. These results suggest that edge plane pyrolytic graphite can be optimally used as the working electrodes in Clark cell devices for low-potential amperometric gas sensing of Cl2. PMID- 15883787 TI - Recent advances in peptide chiral selectors for electrophoresis and liquid chromatography. AB - The application of peptides in chiral separations using techniques such as capillary electrophoresis (CE), electrokinetic capillary chromatography (EKC) and liquid chromatography is the focus of this review. Methods for finding peptide selectors using combinatorial library approaches are discussed, as well as recent advances in the use of peptides as general chiral selectors for electrophoresis and liquid chromatography. One example shows the effectiveness of polymeric dipeptide surfactants as general chiral selectors for electrophoresis. Another example shows the versatility of oligoproline chiral stationary phases, exhibiting resolution for a number of racemic analytes comparable to other well established chiral stationary phases. PMID- 15883788 TI - Retention characteristics of a new butylimidazolium-based stationary phase. AB - A new HPLC stationary phase has been synthesized based on the ionic liquid n butylimidazolium bromide. Imidazolium was covalently immobilized on a silica substrate through an n-alkyl tether and the retention characteristics of the resulting stationary phase were evaluated systematically. Using 28 small aromatic test solutes and reversed phase conditions, the linear solvation energy relationship approach was successfully used to characterize this new phase. The retention characteristics of the test solutes show remarkable similarity with phenyl stationary phases, despite the presence of a positive charge on the new imidazolium phase. Operated in the reversed phase mode, this new stationary phase shows considerable promise for the separation of neutral solutes and points to the potential for a truly multi-modal stationary phase. PMID- 15883789 TI - Liquid chromatographic properties of aromatic sulfur heterocycles on a Pd(II) containing stationary phase for petroleum analysis. AB - Polycyclic aromatic sulfur heterocycles (PASHs) can show very poor reactivities in catalytic hydrodesulfurization processes in refineries, especially those in high-boiling fractions and distillation residues. An insight into the structural features of the most recalcitrant PASHs is essential for developing more efficient catalysts and improving refinery processes. The very high complexities of such mixtures necessitate fractionation of the samples into smaller subsets according to defined criteria. A stationary phase containing a palladium(II) complex was previously shown to be efficient for separating PASHs in lighter petroleum fractions. Here we characterize this ligand exchange chromatographic phase using a large number of sulfur aromatic model compounds that were synthesized for the purpose. In general, compounds containing thiophene rings that are not condensed with other aromatic systems are weakly retained and elute in a first fraction with the polycyclic aromatic hydrocarbons. Thiophene rings condensed with other aromatic rings are more strongly retained and elute in a later fraction with a more polar eluent. If the sulfur is in a non-aromatic ring, the compound is irreversibly retained by the Pd(II) ions. Some steric effects are seen in compounds with alkyl or aryl substituents close to the sulfur atom but in general they do not interfere strongly with the complexation. Thus it seems possible to separate groups of aromatic sulfur compounds according to their complexation properties. For instance, such fractionated samples can be studied much more easily by mass spectrometric techniques. PMID- 15883790 TI - Fabrication of an integrated PDMS microchip incorporating an LED-induced fluorescence device. AB - A microfluidic device with an integrated fluorescence detection system has been developed in order to miniaturize the entire analytical system. A blue or green light-emitting diode (LED) and an optical fiber were mounted in a polydimethylsiloxane-based microchip. The performance of this device was evaluated by microchip electrophoresis. When a green LED was used as the light source, the calibration curve of Sulforhodamine-101 was linear over the range 1 100 microM. The detection limit was found to be 600 nM (240 amol) for a S/N ratio of 3. When using a blue LED, the calibration curve of Fluorescein was linear over the range 0.2-100 microM. The detection limit was estimated to be 120 nM (50 amol) (S/N=3). The detection sensitivity per unit power was comparable to that of LIF. The RSD values for the migration time, peak height and peak area were 0.74, 7.18 and 9.45%, respectively. The integrated microfluidic device was successfully used to determine amino acid derivatives. PMID- 15883791 TI - Genomic cloning and linkage mapping of the Mal d 1 (PR-10) gene family in apple (Malus domestica). AB - Fresh apples can cause birch pollen-related food allergy in northern and central European populations, primarily because of the presence of Mal d 1, the major apple allergen that is cross-reactive to the homologous and sensitizing allergen Bet v 1 from birch. Apple cultivars differ significantly in their allergenicity. Knowledge of the genetic basis of these differences would direct breeding for hypoallergenic cultivars. The PCR genomic cloning and sequencing were performed on two cultivars, Prima and Fiesta, which resulted in 37 different Mal d 1 gDNA sequences. Based on the mapping of sequence-specific molecular markers, these sequences appeared to represent 18 Mal d 1 genes. Sixteen genes were located in two clusters, one cluster with seven genes on linkage group (LG) 13, and the other cluster with nine genes on the homoeologous LG 16. One gene was mapped on LG 6, and one remained unmapped. According to sequence identity, these 18 genes could be subdivided into four subfamilies. Subfamilies I-III had an intron of different size that was subfamily and gene-specific. Subfamily IV consisted of 11 intronless genes. The deduced amino acid sequence identity varied from 65% to 81% among subfamilies, from 82% to 100% among genes within a subfamily, and from 97.5% to 100% among alleles of one gene. This study provides a better understanding of the genetics of Mal d 1 and the basis for further research on the occurrence of allelic diversity among cultivars in relation to allergenicity and their biological functions. PMID- 15883792 TI - Rhg1 alleles from soybean PI 437654 and PI 88788 respond differentially to isolates of Heterodera glycines in the greenhouse. AB - The production of resistant soybean [Glycine max (L.) Merr.] cultivars is the most effective means for controlling losses from soybean cyst nematode (SCN) (Heterodera glycines Ichinohe). The major resistance gene in most SCN resistance sources is rhg1, which has been mapped as a quantitative trait locus onto linkage group G. Our objective was to determine whether the SCN resistance sources PI 437654 and PI 88788 have different functional alleles at rhg1 based on resistance phenotypes. Populations segregating for resistance alleles at rhg1 from both PI 88788 and PI 437654 and at Rhg4, a second SCN resistance gene from PI 437654, were developed. These populations were screened for resistance to the H. glycines inbred isolates PA3 (HG type 7) and TN14 (HG type 1.2.5.7) in the greenhouse and evaluated with molecular markers linked to both rhg1 and Rhg4. Each isolate test was repeated, and the evaluations were done on a single-plant and a line-mean basis in Test 1, and solely on a single-plant basis in Test 2. Across two tests with the TN14 isolate, plants with the PI 437654 allele for a marker linked to rhg1 had significantly (P<0.0001) less SCN reproduction than plants carrying the PI 88788 allele. A marker linked to Rhg4, however, was not significantly associated with resistance to TN14. Across two tests with the PA3 isolate, alleles of rhg1 from both sources gave a resistant reaction, although plants homozygous for the PI 88788 allele had significantly (P<0.05) greater resistance than plants with the PI 437654 allele. The marker allele from PI 437654 linked to Rhg4 was significantly (P<0.0005) associated with greater resistance than the PI 88788 allele in both PA3 tests, and resistance was dominant. There was a significant interaction between alleles at rhg1 and Rhg4 in both PA3 tests. These results suggest that PI 437654 and PI 88788 each have a different functional SCN resistance allele at or close to rhg1. These allelic differences have implications that breeders should consider before incorporation into cultivars. PMID- 15883793 TI - Chromosomal location of a pollen fertility-restoring gene, Rf, for CMS in Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of A. galanthum Kar. et Kir. revealed by genomic in situ hybridization. AB - In a previous study, we developed cytoplasmic male sterile lines of Allium fistulosum possessing the cytoplasm of A. galanthum, a wild species, by continuous backcrossing. Furthermore, we reported the presence of a pollen fertility-restoring gene (Rf) for cytoplasmic male sterility (CMS) in A. fistulosum from segregation of pollen fertility of backcross progenies. In the present study, genomic in situ hybridization (GISH), using genomic DNA of A. galanthum as the probe DNA and that of A. fistulosum as the blocking DNA, was applied to F(1) hybrids between both species and backcross progenies to determine the chromosomal location of the Rf locus. By means of GISH, eight chromosomes from A. galanthum were clearly discriminated from those of A. fistulosum in the F(1) hybrids, and chromosome substitution process through continuous backcrossing was visualized. Interestingly, the chromosome region from A. galanthum, specific to male fertile plants, was detected in one chromosome of BC(4) to BC(7) generations. Based on the karyotype analysis of the male fertile plants, the chromosome was identified as the 5F chromosome. Our results confirm that the Rf locus is located on the 5F chromosome of the male fertile plants. This is the first report that identified the chromosomal location of the pollen fertility restoring gene in A. fistulosum. PMID- 15883794 TI - Fatigue in gynaecological cancer patients: a pilot study. AB - RATIONALE: Fatigue is a frequent complaint of women with cancer. However, the incidence of fatigue has not been well studied, in particular gynaecological cancer, which despite its prevalence has received minimal investigation. GOALS OF WORK: The study aims were (1) to explore the symptoms experienced in a gynaecological cancer population, primarily fatigue and (2) to determine the acceptability of a fatigue questionnaire for use in a longitudinal survey. PATIENTS AND METHODS: Over the course of 1 month, women with gynaecological cancer attending a Regional Cancer Centre completed a demographic and symptom questionnaire and the Multidimensional Fatigue Symptom Inventory-Short Form (MFSI SF). MAIN RESULTS: Of the 32 individuals approached, 30 agreed to participate (mean age, 61 years; the most common treatment received was surgery followed by chemotherapy n=11; mean time from commencement of treatment, including surgery = 3 months). All participants completed the MFSI-SF. Tiredness was the most commonly reported symptom, experienced by 90% of subjects and the most frequently stated worst symptom, reported by 23.3%. Furthermore, 23 of 27 subjects reported that tiredness interfered completely with their daily living. The MFSI-SF mean total fatigue score was 14.4 (SD 15.9), ranging from -15 to 50. The possible total fatigue score ranges from -24 to 96. CONCLUSION: Despite the heterogeneous nature of the group, all participants completed the MFSI-SF. The study suggests that fatigue could be a problem for this population group. Thus, a longitudinal survey using the MFSI-SF to investigate the phenomenon further would appear feasible and justified. PMID- 15883795 TI - [New medications for treatment of systemic mycoses]. AB - The past few years have seen the advent of several new antifungal agents. The echinocandin, caspofungin, has greatly expanded the antifungal armamentarium by providing a cell wall-active agent with candidacidal activity as well as demonstrated clinical efficacy in the therapy of aspergillosis refractory to available therapy. In addition, in clinical trials, caspofungin exhibited efficacy comparable to amphotericin B for invasive and/or fluconazole-resistant Candida infections. According to a randomised trial, voriconazole has added a significantly improved therapeutic option for primary therapy of invasive aspergillosis. Additionally, voriconazole may be used successfully as salvage therapy for other fungal infections, i.e. cryptococcosis. Despite the advances offered by each of these drugs, the morbidity and mortality associated with invasive fungal infections remains high. Considering the adverse effects of the available antifungal agents and the considerable costs for their application, meaningful clinical trials for a precise indication in different clinical situations are urgently needed. PMID- 15883799 TI - Information processing during physical exercise: a chronometric and electromyographic study. AB - Choice reaction time (RT) is shorter when participants perform a choice task at the same time as a sub-maximal exercise than when they are at rest. The purpose of the present study was to determine whether such an exercise affects response execution or whether it alters processes located upstream from the neuro-muscular level. To this end, the electromyographic (EMG) activity of the response agonists was analysed in a between-hand choice RT task performed either concurrently with a pedalling task or at rest. Visual stimulus intensity was also manipulated so as to determine whether exercise further affects early sensory processes. Results shows that exercise affected the time interval elapsing from the onset of the contraction of the response agonists to the mechanical response, thereby indicating that this variable modifies the peripheral motor processes involved in response execution. EMG signal analyses further revealed that the cortico-spinal command is more efficient during exercise than at rest. In addition, exercise was shown to interact with visual stimulus intensity on the time between stimulus and voluntary EMG onset and to increase the critical flicker fusion frequency threshold, thereby indicating that exercise modifies the peripheral sensory processes involved in early sensory operations. The decomposition of RT, with respect to the EMG activity of response agonists, sheds light on the processes affected by exercise and suggests that exercise affects both sensory processes and late motor processes. PMID- 15883800 TI - Posterior parietal negativity preceding self-paced praxis movements. AB - Studies of movement-related cortical potentials (MRCPs) for simple movements have shown a slowly rising negativity (Bereitschaftspotential, or BP) about 2 s prior to movement onset, centered in the bilateral sensorimotor area. However, complex movements may elicit a different temporal and spatial distribution of this pre movement activity. In this study, 64-channel electroencephalography (EEG) was recorded while normal volunteers were asked to perform a simple thumb adduction once every 10--15 s for three 10--15 min blocks. Following this, they were asked to make tool-use movements (hammer, scissor, and screwdriver pantomime) in the same manner. Surface electromyography (EMG) was recorded on the thumb adductor and forearm flexor. MRCP was analyzed for the beginning part of the epoch (from 3.5 s to 1.5 s before EMG onset, with 0.5 s time bins) for differences in the amplitude and spatial distribution of the BP. Significant differences were seen from 3.0 s to 2.0 s before EMG onset, where the amplitude was greater for the more complex movements. On average, negativity began at 3.0 s before onset for praxis movements, and only 1.7 s before onset for thumb adduction. Additionally, the negativity seen for the complex movements had a distribution beginning over the left hemisphere posterior parietal area, whereas, thumb adduction movements had a more anterior distribution, over the bilateral sensorimotor area. The posterior parietal negativity (PPN) suggests that early parietal activity is essential for tool-use movements and is not a part of preparing simple movements. PMID- 15883801 TI - The time-course of ribavirin-provoked changes of basal and AMPH-induced motor activities in rats. AB - The time-course of changes of basal and amphetamine (AMPH)-induced locomotor and stereotypic activities in adult male Wistar rats after a single ribavirin injection was studied. In the first set of experiments, 10, 20 or 30 mg ribavirin/kg body weight (b.w.) were injected i.p. to rats and their basal motor activities were recorded every 10 min for 2 h and compared with those of saline treated controls. In the second set of experiments, the animals were pretreated with ribavirin and 20 min later i.p. injected with AMPH (1.5 mg/kg b.w.). The controls received AMPH 20 min after the saline injection. Motor activity was recorded after the first injection and until 120 min after AMPH administration. Ribavirin did not significantly affect the time-course of either basal locomotor or stereotypic activities. Pretreatment with any of the applied ribavirin doses decreased the AMPH-induced hyperlocomotor response. However, the most pronounced effect was observed with ribavirin doses of 20 mg/kg and 30 mg/kg when administered during the first 10 min and 30 min after the AMPH injection respectively. In contrast, the stereotypic activities of these animals were only slightly changed. These results indicate a different susceptibility of regions in the basal ganglia to ribavirin. PMID- 15883802 TI - Redirection of gaze and switching of attention during rapid stepping reactions evoked by unpredictable postural perturbation. AB - In many situations successful execution of a balance-recovery reaction requires visual information about the environment. In particular, reactions that involve rapid limb movements, such as stepping, must be controlled to avoid obstacles and accommodate other constraints on limb trajectory. However, it is unknown whether the central nervous system can acquire the necessary visuospatial information prior to perturbation onset or must, instead, redirect gaze at the floor during the execution of the stepping reaction. To study this we examined gaze behaviour, during rapid forward-directed stepping reactions triggered by unpredictable platform perturbation, in 12 healthy young adults. We also monitored switching of attention, as inferred from onset of significant error in performing a concurrent visuomotor tracking task. Obstacles and/or step targets were used as constraints, to increase demands for accurate foot movement. Downward gaze shifts towards the floor almost never occurred during stepping reactions when foot motion was unconstrained but did occur more frequently as the demands for accurate foot movement increased. Nonetheless, even in the most challenging condition (target plus obstacle), downward redirection of gaze occurred in less than 40% of the trials, and subjects were commonly well able to clear the obstacle and land the foot on the target without redirecting their gaze towards the floor. An apparent switching of attention, subsequent to perturbation onset, occurred frequently (>80% of trials) in all task conditions, independent of the gaze shifts. The findings indicate that visual fixation of the foot or floor was not essential for accurate control of the foot movement, nor was the apparent switching of attention that followed perturbation onset linked, in any consistent way, to overt changes in visual fixation. Spatial features of the support surface were apparently "remembered" prior to perturbation onset, thereby allowing both vision and attention to be directed to other demands during the execution of the balance reaction. PMID- 15883803 TI - Responses of muscle spindles in feline dorsal neck muscles to electrical stimulation of the cervical sympathetic nerve. AB - Previous studies performed in jaw muscles of rabbits and rats have demonstrated that sympathetic outflow may affect the activity of muscle spindle afferents (MSAs). The resulting impairment of MSA information has been suggested to be involved in the genesis and spread of chronic muscle pain. The present study was designed to investigate sympathetic influences on muscle spindles in feline trapezius and splenius muscles (TrSp), as these muscles are commonly affected by chronic pain in humans. Experiments were carried out in cats anesthetized with alpha-chloralose. The effect of electrical stimulation (10 Hz for 90 s or 3 Hz for 5 min) of the peripheral stump of the cervical sympathetic nerve (CSN) was investigated on the discharge of TrSp MSAs (units classified as Ia-like and II like) and on their responses to sinusoidal stretching of these muscles. In some of the experiments, the local microcirculation of the muscles was monitored by laser Doppler flowmetry. In total, 46 MSAs were recorded. Stimulation of the CSN at 10 Hz powerfully depressed the mean discharge rate of the majority of the tested MSAs (73%) and also affected the sensitivity of MSAs to sinusoidal changes of muscle length, which were evaluated in terms of amplitude and phase of the sinusoidal fitting of unitary activity. The amplitude was significantly reduced in Ia-like units and variably affected in II-like units, while in general the phase was affected little and not changed significantly in either group. The discharge of a smaller percentage of tested units was also modulated by 3-Hz CSN stimulation. Blockade of the neuromuscular junctions by pancuronium did not induce any changes in MSA responses to CSN stimulation, showing that these responses were not secondary to changes in extrafusal or fusimotor activity. Further data showed that the sympathetically induced modulation of MSA discharge was not secondary to the concomitant reduction of muscle blood flow induced by the stimulation. Hence, changes in sympathetic outflow can modulate the afferent signals from muscle spindles through an action exerted directly on the spindles, independent of changes in blood flow. It is suggested that such an action may be one of the mechanisms mediating the onset of chronic muscle pain in these muscles in humans. PMID- 15883804 TI - Electrophysiological correlates of neural plasticity compensating for ischemia induced damage in the hippocampus. AB - Injury to the brain often results in loss of synapses or cell death in the damaged area. Subsequent to the injury, the areas that are not directly affected often exhibit enhanced neuronal plasticity. Although there are many reports of morphological changes resulting from such plasticity, their functional consequences are poorly understood. In this study we examined electrophysiological changes associated with ischemia-induced neurogenesis in the hippocampus, a brain region that is particularly vulnerable but also exceptionally plastic. Transient global ischemia was induced in Sprague-Dawley rats by occlusion of both carotid arteries and a reduction in blood pressure for 12 min. The procedure resulted in delayed cell death in the CA1 field of the hippocampus while the dentate gyrus (DG) was spared. To assess neurogenesis and synaptic changes in parallel we used both hemispheres from each animal. One side was used for immunohistochemistry and the other for in vitro electrophysiological experiments in brain slices. Synaptic field responses and synaptic plasticity (LTP) in perforant path within the DG were reduced by 50% at 10 days after the ischemic injury but recovered at 35 days. Synaptic responses in non-neurogenic CA1 were abolished in parallel with cell death and did not recover. Gamma irradiation applied focally to the head selectively prevented neurogenesis and the synaptic recovery in the DG. These experiments reveal electrophysiological changes associated with reactive neural plasticity in the hippocampus. PMID- 15883805 TI - Response to unexpected target changes during sustained visual tracking in schizophrenic patients. AB - BACKGROUND: Evidence supports an association between liability to schizophrenia and smooth-pursuit eye movement (SPEM) abnormalities. Knowledge of the biological mechanisms of SPEM abnormalities may provide critical insights into the etiology of schizophrenia. SPEM is elicited by sensory motor information from the movement of the object's image on the retina (retinal motion signal) and subsequent extraretinal motion signals. Previous studies suggest that a deficit in predictive responses to extraretinal motion signals may underlie the SPEM phenotype in schizophrenia. Data suggest that at-risk individuals for schizophrenia depend less on extraretinal and more on retinal motion signals to maintain pursuit than healthy individuals. METHODS: We designed a pursuit task that employs unexpected changes in target direction during smooth pursuit. The unpredictable task is unique in that performance is expected to be better if the subject's response is biased towards retinal motion. RESULTS: The study included 23 schizophrenia patients and 22 normal controls. Results showed that schizophrenia patients showed significantly superior performance (i.e. higher smooth pursuit gain) for a brief period after an unexpected change in target direction compared with healthy subjects. CONCLUSION: Findings of superior performance by schizophrenic patients are interesting because they circumvent confounds of generalized deficits. These results provide further evidence of specific deficit in the predictive pursuit mechanism and over-reliance on retinal error signals to maintain pursuit in schizophrenia. PMID- 15883806 TI - Action observation supports effector-dependent learning of finger movement sequences. AB - Practising a motor skill can result in effector-dependent learning (learning that does not transfer from the set of muscles used in training to a new set of muscles). Proceeding from neurophysiological evidence of motor activation during action observation, this study asked whether observational learning, learning through observation of skilled performance, can also be effector-dependent. Adult human participants observed a model's right hand as the model responded to an eight-item sequence in a serial reaction time (SRT) task. Their sequence learning was then compared in two tests with that of controls who had observed the model's right hand responding to random targets during training. All participants performed the SRT task with their right hand in the first test and with their left hand in the second. Evidence of observational learning was obtained in the right hand test but not in the left hand test. This implies that sequence learning based on observation of right hand performance did not transfer to the left hand, and therefore that observational learning can support effector dependent learning of finger movement sequences. A second experiment used the same procedure to assess learning by a group of participants who observed a sequence of response locations only. This group did not observe the model's responses. Results suggested that action observation was necessary for the effector-dependent observational learning demonstrated in Experiment 1. PMID- 15883807 TI - Vision and gaze direction modulate tactile processing in somatosensory cortex: evidence from event-related brain potentials. AB - Several behavioural studies have shown that directing one's gaze at a body part reduces detection speed and enhances discrimination of tactile stimuli at that location. We investigated how vision of a body part stimulated and manipulations of gaze direction affect tactile processing. Participants' gaze was directed to one of their hands, with vision of this hand either available or prevented in different experiments. They had to detect infrequent tactile targets among non targets. Somatosensory event-related brain potentials were recorded in response to stimulation of the hand towards which gaze was directed (G+ trials) and in response to stimulation of the other hand (G- trials). When vision (V+) of the hand gaze was directed at was available (G+V+), an early positivity overlapping with the P45 and N 80 component was observed for G+V+ trials relative to G-V- trials. In contrast, when the hands were occluded from view (V-), an enhanced N 140 component followed by a late negativity was observed for G+V- as compared to G-V- trials. It is suggested that vision of the body part stimulated can modulate processing in primary somatosensory cortex (S1), while effects of gaze direction in the absence of vision of the body part touched are located in higher order somatosensory areas. Such effects of vision and gaze on tactile processing may be mediated by pathways from multimodal brain regions to somatosensory cortex. PMID- 15883808 TI - Cortico-motoneuronal output to intrinsic hand muscles is differentially influenced by static changes in shoulder positions. AB - We investigated whether shoulder position influenced the recruitment properties of the abductor digiti minimi muscle (ADM) and first dorsal interosseous muscle (FDI). ADM and FDI motor evoked potentials (MEPs) in response to transcranial magnetic stimulation (TMS) were obtained in seven healthy volunteers at two different static positions of the shoulder joint (30 degrees adduction vs 30 degrees abduction) while the arm was passively supported at shoulder level (90 degrees in the horizontal plane) and the elbow joint was fixed at 90 degrees . ADM and FDI voluntary activity was also examined during (1) externally paced finger abductions at 2 Hz in the two different shoulder positions (EMG(ADM) and EMG(FDI) was back-averaged time-locked to the end of finger abduction) and (2) maximal voluntary abduction of the little finger and the index finger. Maximal EMG power and force were analysed in the two shoulder positions. H-reflexes from ADM and FDI were also obtained in two subjects. The ADM stimulus-response curve to TMS showed that the slope and plateau level were significantly reduced with the shoulder at 30 degrees abduction. In contrast, the FDI stimulus-response curve to TMS was not influenced by shoulder position. The back-averaged EMG(ADM) showed a significant decrease in peak amplitude and area with the shoulder at 30 degrees abduction, while no change in EMG(FDI) was observed under the same condition. Similarly, maximal EMG(ADM) and force exertion by the little finger were significantly reduced with the shoulder at 30 degrees abduction, while no such effect was observed for FDI. ADM H-reflex, but not FDI, was also decreased with shoulder abduction. These results indicate that the corticospinal pathway to ADM is less accessible to TMS and to voluntary command when the shoulder is placed at 30 degrees abduction. In contrast, activation of FDI, whether by TMS or by volition, is not influenced by shoulder position. This finding suggests that there are differences in the corticospinal innervation to ADM and FDI, possibly due to the different role of these muscles in hand function. PMID- 15883809 TI - Trajectory of contact region on the fingerpad gives the illusion of haptic shape. AB - When one explores a solid object with a fingertip, a contact region is usually defined. When the trajectory of this region on the fingerpad is artificially controlled so as to resemble the trajectory that is normally present while exploring a real object, the experience of shape is created. In order to generate appropriate local deformation trajectories, we built a servo-controlled mechanism that rolled a flat plate on the fingerpad during the manual exploration of virtual surfaces so that the plate was kept tangent to a virtual shape at the point of virtual contact. An experiment was then designed to test which mode of exploration maximized the shape information gain: active versus semi-active exploration, where semi-active exploration is when one hand touches passively and the other moves the target object, and the use of single versus multiple points of contact. We found that subjects were able to perform curvature discrimination at levels comparable to those achieved when using direct manual contact with real objects, and that the highly simplified stimulus provided by the device was a sufficient cue to give the illusion of touching three-dimensional surfaces. PMID- 15883810 TI - Braking of elbow extension in fast overarm throws made by skilled and unskilled subjects. AB - A previous computer simulation study of overarm throws in 2D showed that reversal of elbow torque by antagonist muscle action late in the throw led to increased wrist flexion velocity and to increased ball speeds. We tested the hypothesis that the skill of making fast overarm throws in 3D involves deceleration (braking) of elbow extension before ball release, and that this is an active mechanism. Skilled and unskilled throwers were instructed to throw baseballs at a fast speed. Arm segment angular positions in 3D at 1,000 Hz were recorded with the search-coil technique (which records angular motions). In skilled throws, but not in unskilled throws, there was a period (mean 17 ms) of rapid elbow extension deceleration before ball release. However, there was relatively little biceps EMG activity associated with the very large magnitude of elbow deceleration. This finding and other work suggests that elbow extension deceleration results in part from interaction torques associated with late-occurring shoulder rotations, and only in part from elbow flexor contraction. During the period when elbow extension was decelerating, the forearm in space was undergoing angular acceleration (because of internal rotation at the shoulder) which would be expected to produce a torque at the wrist in the extensor (not flexor) direction. The results show that elbow extension deceleration occurs before ball release in fast (skilled) 3D throws, and that it does not produce forearm angular deceleration. Whether it produces forearm translational deceleration, which could increase wrist flexion velocity, remains to be determined. PMID- 15883811 TI - Factors affecting the size of the detour effect in the kinaesthetic perception of Euclidean distance. AB - Three experiments investigated the mechanisms by which we estimate Euclidean distances on the basis of kinaesthetic cues. In all experiments, blindfolded participants followed straight and curvilinear paths with a stylus. Then, with a straight response movement, they estimated the distance between the end-points of the previously explored path. Experiment 1 was designed to validate the hypothesis-made on the basis of results from a previous study-that errors in the kinaesthetic estimations of distances (detour effect) originate from the difficulty to decompose the displacement vector into relevant and irrelevant components, which would become more severe at points of inflection. Using elliptic paths (no inflections), we demonstrated that errors are indeed reduced considerably. The role of the orientation of the work plane was investigated in Experiment 2 in which the same paths used in our previous study were oriented in the frontal rather than the horizontal plane. The results indicate that the detour effect is independent of the orientation. Moreover, despite the asymmetry that gravity introduces between upward and downward movements, errors in the two directions are almost identical. Experiment 3 addressed two issues. First, we demonstrated that introducing a delay between the exploration of the path and the response did not alter significantly the pattern of errors. By contrast, we demonstrated that errors are severely reduced when the number of paths to be explored is reduced by half. The results of the three experiments are discussed within the context of current theories of sensori-motor coding. PMID- 15883812 TI - Psychophysiology of ejaculatory function and dysfunction. AB - Psychophysiological research has attempted to understand the relationship between subjective psychological experiences (perceptive, affective, cognitive) and physical/physiological components of sexual response, including erection and ejaculation. Such research has enabled the investigation of a number of hypotheses related to the etiology and/or cause of premature ejaculation (PE), with a number of important clarifications resulting. For example, in PE men, genital stimulation (as opposed to erotic stimulation in general) is particularly germane to dysfunctional response. Autonomic (parasympathetic and sympathetic) response patterns may be altered during the sexual response cycle. Anomalous patterns of arousal are probably involved, though not necessarily in ways that had been conceptualized originally (e.g., hyperarousal). And psychosomatic interactions between negative emotion, autonomic response, and rapid ejaculation may sustain the dysfunctional response. The results of this research dispel long standing hypotheses regarding the etiology of PE, help refocus attention to areas of importance, and raise entirely new questions regarding possible causes of PE. PMID- 15883813 TI - Suppressing effects of daily oral supplementation of beta-glucan extracted from Agaricus blazei Murill on spontaneous and peritoneal disseminated metastasis in mouse model. AB - PURPOSE: The Basidiomycete fungus Agaricus blazei Murill has traditionally been used as a health food for the prevention of cancer. METHODS: We examined whether beta-(1-6)-D: -glucan extracted from A. blazei is a potential anticancer agent in an in vitro and in vivo animal model. RESULTS: Here we show that (1) beta-glucan had cytotoxic effect against human ovarian cancer HRA cells, but not against murine Lewis lung cancer 3LL cells, in vitro; (2) beta-glucan promotes p38 MAPK activity for suppressing HRA cell proliferation and amplifying the apoptosis cascade; (3) beta-glucan stimulates translocation of the proapoptotic protein, Bax, from the cytosol to mitochondria, cytochrome c release, and subsequent caspase-9 activation; (4) treatment with SB203580, a p38 MAPK-specific inhibitor, suppresses beta-glucan-induced effects, indicating that activation of p38 MAPK is involved in the suppression of cell proliferation and mitochondrial activation mediated cell death pathway; (5) in mice, oral supplementation with beta-glucan reduces pulmonary metastasis of 3LL cells and peritoneal disseminated metastasis of HRA cells and inhibits the growth of these metastatic tumors in lung or peritoneal cavity, in part, by suppressing uPA expression; and (6) in an in vivo experimental metastasis assay, however, the oral supplementation with beta-glucan after i.v. tumor cell inoculation did not reduce the number of lung tumor colonies. CONCLUSION: Treatment with beta-glucan may be beneficial for cancer patients with or at risk for metastasis. The beta-glucan-dependent signaling pathways are critical for our understanding of anticancer events and development of cancer therapeutic agents. PMID- 15883814 TI - Macrophage level is not affected by monocyte chemotactic protein-1 in invasive ductal breast carcinoma. AB - PURPOSE AND METHOD: Monocyte chemotactic protein-1 (MCP-1) is a chemokine involved in the macrophage infiltration of tumor tissue. Tumor-associated macrophages (TAMs) are a population of mononuclear phagocytic cells that can have a complex function in tumor biology. The aim of this study was to determine the possible correlation between parenchymal MCP-1 expression and TAM level by immunohistochemical analysis of 97 invasive ductal breast carcinomas, not otherwise specified (NOS), and to investigate their relation with tumor size, histological grade, mitotic activity index (MAI) and lymph node status. Secondly, the MCP-1 mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) in eight samples of normal breast tissue and 27 samples of invasive breast carcinomas and compared with TAMs. RESULTS: MCP-1 immunoreactivity was present in tumor cells (17/97), but also in TAMs, fibroblasts and endothelial cells. The statistical analysis did not show a significant correlation between MCP-1 expression in tumoral epithelium and tumor size, histological grade, MAI, lymph node status or TAMs. The results of RT-PCR showed that, in all cases of breast carcinomas (27/27) and the majority of normal breast tissues (7/8), the number of detected MCP-1 cDNA copies was above the detection limit. However, carcinomas showed higher levels of MCP-1 mRNA than normal breast tissue. Nevertheless, the statistical analysis did not find a significant correlation between MCP-1 expression and macrophage infiltrations. CONCLUSION: These results indicate that MCP-1 is probably not the only and/or crucial factor involved in macrophage attraction to tumor locus in breast carcinoma. PMID- 15883815 TI - Polymorphism in the manganese superoxide dismutase (MnSOD) gene and risk of breast cancer in young women. AB - PURPOSE: Manganese superoxide dismutase (MnSOD) is one of the major enzymes implicated in the cellular defence against reactive oxygen species. Low expression of MnSOD has been observed in different cancer tissues and several reports have shown that overexpression of MnSOD inhibits growth in various human cancer cells. These observations suggest that MnSOD is involved in carcinogenesis. A polymorphism (Ala-9Val) in the mitochondrial targeting sequence (MTS) of the MnSOD gene has been proposed to affect protein localization and thereby influence cellular defence against superoxide radicals. METHODS: In the present case-control study, including 118 early onset breast cancer patients (1000 kcal/shift) was 177 g lower than that of those whose mothers worked out of greenhouse (light work <700 kcal/shift) (p = 0.05). Mothers who during work in greenhouse were potentially exposed to RD pesticides, delivered infants with birth weight lower by about 70 g. than infants' mothers not working at places where pesticides RD were applied, but these findings were not statistically significant. CONCLUSION: Our results indicate that infants of mothers performing heavy work inside greenhouse during pregnancy had lower mean birth weight than infants of mothers working out of greenhouse. No similar effects of current exposure to pesticides was observed. PMID- 15883818 TI - Occupational hand-arm vibration syndrome in Korea. AB - OBJECTIVES: It is suspected that there is a large number of patients suffering from hand-arm vibration syndrome (HAVS) in Korea. However, no cases have been reported since 1992. This study was conducted to identify HAVS cases and determine the characteristics of the syndrome. METHODS: In April 2001, the Busan, Ulsan, and Gyeong-Nam Province Occupational Disease Surveillance System (BUGODSS) was established to identify work-related HAVS and other occupational diseases. In the 2 years of this project, occupational physicians from five hospitals in these provinces collected information by way of interviews and questionnaires during mandatory occupational medical examinations. Among the initial 189 suspected HAVS cases, 58 volunteers were given cold-water provocation tests in order to diagnose the vascular component of the disorder. RESULTS: One hundred fifty-four approximately occupational HAVS cases were identified from ca. 21,000 workers. One hundred fifty about of these cases were male. The cases were most often found in workers from the shipbuilding industry, and the grinder was the most common source of vibration exposure. Cases of sensorineural disorder (SD) were more common than cases of vascular disorder (VD). The mean values of the finger skin temperature and its recovery rate at 5 min and 10 min after cold-water provocation were significantly lower in the group with the VD than in the group with the SD. CONCLUSIONS: We identified 154 occupational HAVS cases, although no cases have been reported during the occupational medical examinations mandated by the state. The majority of the cases were in workers that used grinders in the shipbuilding industry. We determined that peripheral VD and peripheral SD can progress independently of each other. We conclude that exposure to hand transmitted vibration (HTV) and HAVS cases are common in shipbuilding industry in Korea. The recovery rate of finger skin temperature after cold-water provocation is one of the useful methods for diagnosing the vascular component of HAVS. PMID- 15883819 TI - Pharmacokinetics of imatinib mesylate in end stage renal disease. A case study. AB - AIM: To evaluate the pharmacokinetics of imatinib mesylate (Glivec) and its main metabolite (CGP74588) in a patient with end stage renal disease on hemodialysis and compare it with published data from subjects with normal renal function. PATIENTS AND METHODS: Serial blood samples were collected over a 2-weeks period in a patient who was receiving daily 400 mg oral imatinib mesylate for the treatment of a gastrointestinal stromal tumor metastatic to the liver while on hemodialysis. Plasma levels of imatinib and CGP74588 were determined by a liquid chromatography-tandem mass spectrometry assay. RESULTS: The pharmacokinetic values for imatinib and CGP74588, respectively, were: maximum concentration (3,340 and 781 ng/ml), time to maximum concentration (2 h), half-life (18.2 and 34.0 h), area under the curve (53.9 and 14.8 microg.h/ml), and trough concentration (1,540 and 508 ng/ml) for at least 24 h. All obtained values fell within the range of values of imatinib and its metabolite obtained in patients with normal renal function. Dialysis courses were not found to intervene with plasma kinetics of the study drug. CONCLUSIONS: Our results indicate that the pharmacokinetics of imatinib and its metabolite CGP74588 do not change in patients with end stage renal disease on hemodialysis. Thus, the standard dose of imatinib can be safely administered to patients on hemodialysis, and probably with renal failure, at any stage. PMID- 15883820 TI - Preclinical pharmacology of 2-methoxyantimycin A compounds as novel antitumor agents. AB - PURPOSE: The present study was designed to determine pharmacological and biochemical properties of 2-methoxyantimycin A analogs (OMe-A1, OMe-A2, OMe-A3, and OMe-A5), which are novel antitumor compounds, and provide a basis for future pharmaceutical development, preclinical evaluation, and clinical trials. METHODS: A high-performance liquid chromatography (HPLC) method was established and employed to assess the biostability of these analogs and to determine their pharmacokinetic properties in mice and rats. RESULTS: In vitro biostability of the 2-methoxyantimycin analogs was esterase-dependent, compound-dependent, and species-dependent. In the absence of esterase inhibitors, all of the analogs were relatively unstable. Stability was greater, however, in human and dog plasma than in rat and mouse plasma. In the presence of esterase inhibitors, OMe-A1 was stable at 37 degrees C for 60 min in mouse and rat plasma, moderately stable in human plasma, and unstable in dog plasma. OMe-A2 was generally stable in all types of plasma. OMe-A3 was stable in dog and rat plasma, but not in human or mouse plasma. OMe-A5 was stable in human and dog plasma, but not in mouse or rat plasma. Each of these analogs was highly bound to plasma proteins. Of S9 fractions from four species, human S9 was least efficient in metabolizing OMe-A3. Following an intravenous dose of OMe-A1 in mice, plasma levels decreased rapidly, with an initial half-life of 2.7 min and a terminal half life of 34 min. Following an intraperitoneal dose in mice, plasma levels decreased less rapidly with a terminal half-life of 215 min. Following an intravenous dose of OMe-A1 or OMe-A3 in rats, plasma levels decreased more rapidly with initial half-lives of about 1.0 min. At an equivalent dose, OMe-A3 had a faster clearance than OMe-A1. CONCLUSIONS: For 2-methoxyantimycin A analogs, species differences in biostability, metabolism, and pharmacokinetics may be pertinent in assessing their pharmacological and toxicological profiles and antitumor activity in humans. PMID- 15883821 TI - Genistein-induced apoptosis via Akt signaling pathway in anaplastic large-cell lymphoma. AB - More than half of anaplastic large-cell lymphoma (ALCL) are associated with chromosomal translocation t(2;5)(p23;q35) that leads to the expression of nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) oncoprotein. NPM-ALK activates the antiapoptotic phosphatidylinositol-3 kinase/Akt (PI3K/Akt) signaling pathway, which plays a critical role in cell survival and apoptosis. Inhibition of the PI3K/Akt pathway has been considered as a therapeutic target for cancer where PI3K/Akt activation is a causative factor. Genistein, a natural isoflavonoid found in soy products, has been shown to inhibit cell growth and induce apoptosis in a wide variety of cell lines. Here, we demonstrated that treatment of two t(2;5) ALCL cell lines, SUDHL-1 and Karpas299, with genistein induced apoptosis in a time- and dose-dependent manner. Concurrently, these cells exhibited a decrease in Akt protein levels and subsequent downregulation of Akt activity (Akt phosphorylation). Furthermore, genistein treatment induced mitochondrial membrane potential change, caspase-3 activation and PARP cleavage. From these results, we conclude that inhibition of the Akt signaling pathway and induction of apoptosis by genistein could be used as a new treatment modality for the prevention and/or treatment of t(2;5) ALCL and other hematopoietic malignancies. PMID- 15883822 TI - Soft-tissue reconstruction after meningococcal septicemia using a posterior tibial artery perforator flap in a 6-year-old boy. AB - Fulminant meningococcemia can have life-threatening as well as limb-threatening complications. However, unlike other types of peripheral gangrene, areas of necrosis do not follow known anatomic vascular patterns. Instead, irregular and patchy areas of necrosis are found to exist adjacent to viable tissues. With improved critical care, more patients survive the initial insult and are referred for reconstructive procedures. In this case, a 6-year boy was diagnosed with meningococcemia-induced purpura fulminans. After stabilization, he was transferred to our facility for management of open wounds of both lower extremities and the left elbow. Soft-tissue coverage was accomplished after multiple debridements using various flaps. In particular, a defect of the right midtibia was reconstructed using a posterior tibial artery perforator-based flap with excellent results. This is the first time such a flap has been reported in association with meningococcemia in a patient of this age. PMID- 15883823 TI - Associated anomalies in children with congenital solitary functioning kidney. AB - Congenital solitary functioning kidney (CSFK) is a relatively common renal malformation and in children is frequently complicated by anomalies of the ipsilateral genital organs and occasionally by anomalies of other organs. The aim of this study was to determine the incidence of associated urological, cardiac, gastrointestinal, hematological, neurological, skeletal, and other congenital malformations in children with CSFK. We retrospectively reviewed 87 consecutive cases of CSFK diagnosed at our hospital between 1995 and 2003. There were 45 boys and 42 girls, whose ages at diagnosis ranged from newborn to 16 years (mean 4.67+/-4.48 years). In all patients, CSFK was diagnosed by abdominal ultrasound and confirmed by radionuclide studies. In 46 patients (53%) the left kidney was absent, and in 41 patients (47%) the right kidney was absent. Overall associated anomalies were detected in 52 of the 87 children (60%) with CSFK. Urological anomalies were most common, with an incidence of 37% (32/87). Nonurological anomalies were detected in 38 children (44%) with CSFK; these included cardiac anomalies in 13, gastrointestinal anomalies in eight, hematological anomalies in five, neurological anomalies in three, and other organ anomalies in nine. In our study, more than half of the patients with CSFK had associated anomalies. For this reason we recommend abdominal ultrasound and voiding cystourethrogram for early recognition of urological anomalies and a careful physical examination for other organ anomalies in patients with CSFK. PMID- 15883824 TI - A candidate autonomous version of the wheat MITE Hikkoshi is present in the rice genome. AB - A miniature inverted-repeat transposable element (MITE), designated as Hikkoshi, was previously identified in the null Wx-A1 allele of Turkish bread wheat lines. This MITE is 165 bp in size and has 12-bp terminal inverted repeats (TIRs) flanked by 8-bp target site duplications (TSDs). Southern and PCR analyses demonstrated the presence of multiple copies of Hikkoshi in the wheat genome. Database searches indicated that Hikkoshi MITEs are also present in barley, rice and maize. A 3.4-kb element that has Hikkoshi-like TIRs flanked by 8-bp TSDs has now been identified in the rice genome. This element shows high similarity to the 5' subterminal region of the wheat Hikkoshi MITE and contains a transposase (TPase) coding region. The TPase has two conserved domains, ZnF_TTF and hATC, and its amino acid sequence shows a high degree of homology to TPases encoded by Tip100 transposable elements belonging to the hAT superfamily. We designated the 3.4-kb element as OsHikkoshi. Several wheat clones deposited in EST databases showed sequence similarity to the TPase ORF of OsHikkoshi. The sequence information from the TPase of OsHikkoshi will thus be useful in isolating the autonomous element of the Hikkoshi system from wheat. PMID- 15883825 TI - Identification of residues in the WD-40 repeat motif of the F-box protein Met30p required for interaction with its substrate Met4p. AB - The SCF family of ubiquitin-ligases consists of a common core machinery, namelySkp1p, Cdc53p, Hrt1p, and a variable component, the F-box protein that is responsible for substrate recognition. The F-box motif, which consists of approximately 40 amino acids, connects the F-box protein to the core ubiquitin ligase machinery. Distinct SCF complexes, defined by distinct F-box proteins, target different substrate proteins for proteasome-dependent degradation. As part of the SCF(Met30p) complex, the F-box protein Met30p selects the substrate Met4p, a transcriptional activator for MET biosynthetic genes that mediate sulfur uptake and biosynthesis of sulfur containing compounds. When cells are grown in the absence of methionine, Met4p evades degradation by the SCF(Met30p) complex and activates the MET biosynthetic pathway. However, overproduction of Met30p represses MET gene expression and induces methionine auxotrophy in an otherwise methionine prototrophic strain. Here we demonstrate that overproduction of the C terminal portion of Met30p, which is composed almost entirely of seven WD-40 repeat motifs, is necessary and sufficient to induce methionine auxotrophy and complement the temperature sensitive (ts) met30-6 mutation. Furthermore, we show that this region of Met30p is important for binding Met4p and that mutations that disrupt this interaction prevent both the induction of methionine auxotrophy and complementation of the met30-6 mutation. These assays have been exploited to identify residues that are important for the interaction of Met30p with its substrate. Since the C-terminal domain of Met30p lacks the F-box and cannot support the ubiquitination of Met4p, our results indicate that the recruitment of Met4p to the SCF(Met30p) complex itself results in inactivation of Met4p, independently of its ubiquitination. PMID- 15883826 TI - Characterization of Na, K-ATPase genes in Atlantic salmon (Salmo salar) and comparative genomic organization with rainbow trout (Oncorhynchus mykiss). AB - A combination of molecular and in silico approaches was employed to assemble a survey of Na, K-ATPase genes contained in the ancestrally tetraploid genome of the Atlantic salmon (Salmo salar). Molecular characterization of genomic clones coding for the alpha subunit revealed two single genes (alpha1a and alpha2) and two pairs of presumably homeologous genes (alpha1b/i-ii and alpha1c/i-ii). Each of the six genes showed high sequence similarity to isoforms previously isolated from rainbow trout and extensive structural differences relative to putative orthologs in the human genome. In silico analysis of expressed sequence tag (EST) collections indicated that at least five alpha (alpha1a, alpha1b, alpha1c, alpha2, and alpha3) and four beta (beta1a, beta1b, beta2, and beta3b) subunit isoforms are expressed in Atlantic salmon. Meiotic linkage analysis further showed that Na, K-ATPase genes are dispersed throughout the salmon genome, with the exception of two multigene clusters on linkage groups AS-22 and AS-28. Duplicate gene copies for the isoform alpha1b were assigned to linkage groups with multiple homeologous anchors (AS-22 and AS-23), while beta2 duplicates suggested a new homeologous affinity between AS-05 and AS-28. In addition, the comparison of linkage arrangements with rainbow trout also showed that the genomic organization of Na, K-ATPase genes is consistent with the evolutionary conservation of syntenic chromosome regions between these species. PMID- 15883827 TI - Coexistence of pleomorphic xanthoastrocytoma with Sturge-Weber syndrome: MRI features. AB - Pleomorphic xanthoastrocytoma is a rare, cortically based, partially cystic astroglial tumour with a strongly enhancing solid mural nodule. We report an 11 year-old boy with a pleomorphic xanthoastrocytoma in the contralateral hemisphere to that affected by Sturge-Weber syndrome. This unique case supports the association between Sturge-Weber syndrome and astrocytomas. Patients with Sturge Weber syndrome who experience unexpected neurological symptoms should be evaluated with MRI. PMID- 15883828 TI - Which MRI sequence of the spine best reveals bone-marrow metastases of neuroblastoma? AB - BACKGROUND: MRI is an effective tool in evaluating bone marrow metastases. However, no study has defined which MRI sequences or image characteristics best correlate with bone-marrow metastases in neuroblastoma. OBJECTIVE: To identify and refine MRI criteria and sequence selection for the diagnosis of bone-marrow metastases in children with neuroblastoma. MATERIALS AND METHODS: Ninety-one children (mean age: 3.2 years; standard deviation: 2.8 years) enrolled in the RDOG IV study participated in our study. Forty-five children had bone metastases determined by bone-marrow aspiration or biopsy (n=4), radionuclide imaging (n=2), or both (n=39). Spine lesions were characterized using coronal T1-weighted (T1W) sagittal short tau inversion recovery (STIR) and coronal gadolinium-enhanced T1 weighted (GAD) MR sequences. Contingency table analysis was performed to determine which MRI sequences and characteristics were associated with metastases. The MRI criteria for metastatic disease were then developed for each imaging sequence. The sensitivity, specificity, predictive values, and accuracy of these criteria were determined for the whole group, children younger than 12 months old, and children 12 months and older. RESULTS: The MR characteristics that had significant (Por=5 mm) as detected by abdominal CT in children with a low likelihood for mesenteric lymphadenopathy. MATERIALS AND METHODS: During a 14 month period, we identified all non-contrast abdominal CT examinations performed at a tertiary care pediatric hospital for evaluation of suspected or known renal stones. Two radiologists reviewed the examinations and recognized all enlarged MLN, measured the short-axis diameter, and noted the quadrant location. RESULTS: Sixty-one children were identified who met entry criteria; mean age was 10.7 years (range 1.1-17.3 years). Enlarged MLN were found in 33 (54%) of the 61 children; the largest enlarged MLN was most frequently in the right lower quadrant (RLQ) (29 of 33, 88%). Seventeen of the 61 children (28%) had three or more enlarged MLN; all were in the RLQ. The largest short-axis diameter measured was 10 mm. SUMMARY: MLN with a short-axis diameter of >5-10 mm are commonly found on abdominal CT examination of children with a low likelihood for mesenteric lymphadenopathy and should be considered a non-specific finding. A short-axis diameter of 8 mm might better define the upper limit of normal mesenteric lymph node size in children. PMID- 15883830 TI - Phytoremediation of toxic trace elements in soil and water. AB - Toxic heavy metals and metalloids, such as cadmium, lead, mercury, arsenic, and selenium, are constantly released into the environment. There is an urgent need to develop low-cost, effective, and sustainable methods for their removal or detoxification. Plant-based approaches, such as phytoremediation, are relatively inexpensive since they are performed in situ and are solar-driven. In this review, we discuss specific advances in plant-based approaches for the remediation of contaminated water and soil. Dilute concentrations of trace element contaminants can be removed from large volumes of wastewater by constructed wetlands. We discuss the potential of constructed wetlands for use in remediating agricultural drainage water and industrial effluent, as well as concerns over their potential ecotoxicity. In upland ecosystems, plants may be used to accumulate metals/metalloids in their harvestable biomass (phytoextraction). Plants can also convert and release certain metals/metalloids in a volatile form (phytovolatilization). We discuss how genetic engineering has been used to develop plants with enhanced efficiencies for phytoextraction and phytovolatilization. For example, metal-hyperaccumulating plants and microbes with unique abilities to tolerate, accumulate, and detoxify metals and metalloids represent an important reservoir of unique genes that could be transferred to fast-growing plant species for enhanced phytoremediation. There is also a need to develop new strategies to improve the acceptability of using genetically engineered plants for phytoremediation. PMID- 15883831 TI - Multicomponent floral signals elicit selective foraging in bumblebees. AB - Flower constancy, or the tendency of individual pollinators to visit sequentially a single flower type even when other equally rewarding types are available, has important implications for animal-pollinated plants. Yet, the proximal reason for the behaviour still remains poorly understood. Here I show that bumblebees visiting equally rewarding flowers that differ in size and odour are more flower constant and less efficient (visited fewer flowers per minute) than bees visiting flowers that differ in size only and odour only. These results are consistent with the view that flower constancy in pollinators is related to their inability to perceive, process or recall multicomponent floral signals. I discuss these findings in the context of pollinator behavioural mechanisms and the evolution of floral diversity. PMID- 15883832 TI - Characterization and dynamic analysis of Arabidopsis condensin subunits, AtCAP-H and AtCAP-H2. AB - Condensin complexes are thought to play essential roles in mitotic chromosome assembly and segregation in eukaryotes. To date, two condensin complexes (condensin I and II) have been identified. Both complexes contain two structural maintenance of chromosome (SMC) subunits and three non-SMC subunits. In plants, little is known about the localization and function of all the condensin subunits. Here, we report the analyses on the localization of a non-SMC subunit of Arabidopsis condensin I and II, AtCAP-H, and AtCAP-H2, respectively. Our study indicated that localization of AtCAP-H and AtCAP-H2 is dynamically changed through the mitotic cell cycle using GFP-tagged AtCAP-H and AtCAP-H2 in tobacco cultured cells. They are localized at mitotic chromosomes from prometaphase to telophase. However, their localization in interphase is quite different. AtCAP-H was mainly found in the cytoplasm whereas AtCAP-H2 was mainly found in a nucleolus. It is revealed using GFP-tagged deletion mutant s of AtCAP-H that the kleisin-gamma middle domain (GM domain) is a unique domain only in AtCAP-H, responsible for chromosomal localization. We propose that the GM domain of CAP-H is essential for its chromosomal localization at mitosis and thus proper function of CAP-H. Differences in localization of AtCAP-H and AtCAP-H2 at interphase also suggest their functional differentiation. PMID- 15883833 TI - Aquaporins in poplar: what a difference a symbiont makes! AB - The formation of ectomycorrhizas, a tight association between fine roots of trees and certain soil fungi, improves plant nutrition in a nutrient-limited environment and may increase plant survival under water stress conditions. To investigate the impact of mycorrhiza formation on plant water uptake, seven genes coding for putative water channel proteins (aquaporins) were isolated from a poplar ectomycorrhizal cDNA library. Four out of the seven genes were preferentially expressed in roots. Mycorrhiza formation resulted in an increased transcript level for three of these genes, two of which are the most prominently expressed aquaporins in roots. When expressed in Xenopus laevis oocytes, the corresponding proteins of both genes were able to transport water. Together, these data indicate, that the water transport capacity of the plasma membrane of root cells is strongly increased in mycorrhized plants. Measurements of the hydraulic conductance of intact root systems revealed an increased water transport capacity of mycorrhized poplar roots. These data, however, also indicate that changes in the properties of the plasma membrane as well as those of the apoplast are responsible for the increased root hydraulic conductance in ectomycorrhizal symbiosis. PMID- 15883834 TI - Flower proteome: changes in protein spectrum during the advanced stages of rose petal development. AB - Flowering is a unique and highly programmed process, but hardly anything is known about the developmentally regulated proteome changes in petals. Here, we employed proteomic technologies to study petal development in rose (Rosa hybrida). Using two-dimensional polyacrylamide gel electrophoresis, we generated stage-specific (closed bud, mature flower and flower at anthesis) petal protein maps with ca. 1,000 unique protein spots. Expression analyses of all resolved protein spots revealed that almost 30% of them were stage-specific, with ca. 90 protein spots for each stage. Most of the proteins exhibited differential expression during petal development, whereas only ca. 6% were constitutively expressed. Eighty-two of the resolved proteins were identified by mass spectrometry and annotated. Classification of the annotated proteins into functional groups revealed energy, cell rescue, unknown function (including novel sequences) and metabolism to be the largest classes, together comprising ca. 90% of all identified proteins. Interestingly, a large number of stress-related proteins were identified in developing petals. Analyses of the expression patterns of annotated proteins and their corresponding RNAs confirmed the importance of proteome characterization. PMID- 15883835 TI - Plant cell wall polysaccharide biosynthesis: real progress in the identification of participating genes. PMID- 15883836 TI - Identification of the form of Cd in the leaves of a superior Cd-accumulating ecotype of Thlaspi caerulescens using 113Cd-NMR. AB - Thlaspi caerulescens (Ganges ecotype) is a known Cd hyperaccumulator, however, the ligands which coordinate to Cd ions in the leaves have not been identified. In the present study, the chemical form of Cd was investigated by using 113Cd nuclear magnetic resonance (NMR) spectroscopy. Plants were grown hydroponically with a highly enriched 113Cd stable isotope. Measurements of 113Cd-NMR with intact leaves showed a signal at the chemical shift of around -16 ppm. Crude leaf sap also gave a similar chemical shift. Purification by gel filtration (Sephadex G-10), followed by cationic and anionic exchange chromatography, showed that Cd occurred only in the anionic fraction, which gave the same chemical shift as intact leaves. Further purification of the anionic fraction, combined with 113Cd- and 1H-NMR studies, revealed that only the fraction containing malate showed a chemical shift similar to the intact leaves. These results indicate that Cd was coordinated mainly with malate in the leaves of T. caerulescens. The malate concentration in the leaves was not affected by increasing Cd concentration in the solution, suggesting that malate synthesis is not induced by Cd. Because the Cd-malate complex is relatively weak, we suggest that the complex forms inside the vacuoles as a result of an efficient tonoplast transport of Cd and a constitutively high concentration of malate in the vacuoles, and that the formation of the Cd-malate complex may lead to a decrease of subsequent Cd efflux to the cytoplasm. PMID- 15883837 TI - SNPs in the neural cell adhesion molecule 1 gene (NCAM1) may be associated with human neural tube defects. AB - Neural tube defects (NTDs) are common birth defects, occurring in approximately 1/1,000 births; both genetic and environmental factors are implicated. To date, no major genetic risk factors have been identified. Throughout development, cell adhesion molecules are strongly implicated in cell-cell interactions, and may play a role in the formation and closure of the neural tube. To evaluate the role of neural cell adhesion molecule 1 (NCAM1) in risk of human NTDs, we screened for novel single-nucleotide polymorphisms (SNPs) within the gene. Eleven SNPs across NCAM1 were genotyped using TaqMan. We utilized a family-based approach to evaluate evidence for association and/or linkage disequilibrium. We evaluated American Caucasian simplex lumbosacral myelomeningocele families (n=132 families) using the family based association test (FBAT) and the pedigree disequilibrium test (PDT). Association analysis revealed a significant association between risk for NTDs and intronic SNP rs2298526 using both the FBAT test (P=0.0018) and the PDT (P=0.0025). Using the HBAT version of the FBAT to look for haplotype association, all pairwise comparisons with SNP rs2298526 were also significant. A replication study set, consisting of 72 additional families showed no significant association; however, the overall trend for overtransmission of the less common allele of SNP rs2298526 remained significant in the combined sample set. In addition, we analyzed the expression pattern of the NCAM1 protein in human embryos, and while NCAM1 is not expressed within the neural tube at the time of closure, it is expressed in the surrounding and later in differentiated neurons of the CNS. These results suggest variations in NCAM1 may influence risk for human NTDs. PMID- 15883838 TI - Anatomy of a founder effect: myotonic dystrophy in Northeastern Quebec. AB - Founder effects are largely responsible for changes in frequency profiles of genetic variants in local populations or isolates. They are often recognized by elevated incidence of certain hereditary disorders as observed in regions of Charlevoix and Saguenay-Lac-Saint-Jean (SLSJ) in Northeastern Quebec. Dominantly transmitted myotonic dystrophy (DM1) is highly prevalent in SLSJ where its carrier rate reaches 1/550, compared with 1/5,000 to 1/50,000 elsewhere. To shed light on the origin of DM1 in this region, we have screened 50 nuclear DM1 families from SLSJ and studied the genetic variation in a 2.05 Mb (2.9 cM) segment spanning the site of the expansion mutation. The markers analyzed included 22 biallelic SNPs and two microsatellites. Among 50 independent DM1 chromosomes, we distinguished ten DM1-associated haplotypes and grouped them into three haplotype families, A, B and C, based on the relevant extent of allele sharing between them. To test whether the data were consistent with a single entry of the mutation into SLSJ, we evaluated the age of the founder effect from the proportion of recombinant haplotypes. Taking the prevalent haplotype A1_21 (58%) as ancestral to all the disease-associated haplotypes in this study, the estimated age of the founder effect was 19 generations, long predating the colonization of Nouvelle-France. In contrast, considering A1_21 as ancestral to the haplotype family A only, yielded the estimated founder age of nine generations, consistent with the settlement of Charlevoix at the turn of 17th century and subsequent colonization of SLSJ. We conclude that it was the carrier of haplotype A (present day carrier rate of 1/730) that was a "driver" of the founder effect, while minor haplotypes B and C, with corresponding carrier rates of 1/3,000 and 1/10,000, respectively, contribute DM1 to the incidence level known in other populations. Other studies confirm that this might be a general scenario in which a major "driver" mutation/haplotype issued from a founder effect is found accompanied by distinct minor mutations/haplotypes occurring at background population frequencies. PMID- 15883839 TI - Molecular and genealogical characterization of the R1443X BRCA1 mutation in high risk French-Canadian breast/ovarian cancer families. AB - The Quebec population contains about six-million French Canadians, descended from the French settlers who colonized "Nouvelle-France" between 1608 and 1765. Although the relative genetic contribution of each of these founders is highly variable, altogether they account for the major part of the contemporary French Canadian gene pool. This study was designed to analyze the role of this founder effect in the introduction and diffusion of the BRCA1 recurrent R1443X mutant allele. A highly conserved haplotype, observed in 18 French-Canadian families and generated using 17 microsatellite markers surrounding the BRCA1 locus, supports the fact that the R1443X mutation is a founder mutation in the Quebec population. We also performed haplotyping analysis of R1443X carriers on 19 other families from seven different nationalities; although the same alleles are shared for three markers surrounding the BRCA1 gene, distinct haplotypes were obtained in four families, suggesting multiple origins for the R1443X mutation. Ascending genealogies of the 18 French Canadian families and of controls were reconstructed on an average depth of 10 generations. We identified the founder couple with the highest probability of having introduced the mutation in the population. Based on the descending genealogy of this couple, we detected the presence of geographical concentration in the diffusion pattern of the mutation. This study demonstrates how molecular genetics and demogenetic analyses can complement each other to provide findings that could have an impact on public health. Moreover, this approach is certainly not unique to breast cancer genetics and could be used to understand other complex traits. PMID- 15883840 TI - Molecular characterisation of the pericentric inversion that distinguishes human chromosome 5 from the homologous chimpanzee chromosome. AB - Human and chimpanzee karyotypes differ by virtue of nine pericentric inversions that serve to distinguish human chromosomes 1, 4, 5, 9, 12, 15, 16, 17, and 18 from their chimpanzee orthologues. In this study, we have analysed the breakpoints of the pericentric inversion characteristic of chimpanzee chromosome 4, the homologue of human chromosome 5. Breakpoint-spanning BAC clones were identified from both the human and chimpanzee genomes by fluorescence in situ hybridisation, and the precise locations of the breakpoints were determined by sequence comparisons. In stark contrast to some other characterised evolutionary rearrangements in primates, this chimpanzee-specific inversion appears not to have been mediated by either gross segmental duplications or low-copy repeats, although micro-duplications were found adjacent to the breakpoints. However, alternating purine-pyrimidine (RY) tracts were detected at the breakpoints, and such sequences are known to adopt non-B DNA conformations that are capable of triggering DNA breakage and genomic rearrangements. Comparison of the breakpoint region of human chromosome 5q15 with the orthologous regions of the chicken, mouse, and rat genomes, revealed similar but non-identical syntenic disruptions in all three species. The clustering of evolutionary breakpoints within this chromosomal region, together with the presence of multiple pathological breakpoints in the vicinity of both 5p15 and 5q15, is consistent with the non random model of chromosomal evolution and suggests that these regions may well possess intrinsic features that have served to mediate a variety of genomic rearrangements, including the pericentric inversion in chimpanzee chromosome 4. PMID- 15883841 TI - Identifying nineteenth century genealogical links from genotypes. AB - We have developed a likelihood method to identify moderately distant genealogical relationships from genomewide scan data. The aim is to compare the genotypes of many pairs of people and identify those pairs most likely to be related to one another. We have tested the algorithm using the genotypes of 170 Tasmanians with multiple sclerosis recruited into a haplotype association study. It is estimated from genealogical records that approximately 65% of Tasmania's current population of 470,000 are direct descendants of the 13,000 female founders living in this island state of Australia in the mid-nineteenth century. All cases and four to five relatives of each case have been genotyped with microsatellite markers at a genomewide average density of 4 cM. Previous genealogical research has identified 51 pairwise relationships linking 56 of the 170 cases. Testing the likelihood calculation on these known relative pairs, we have good power to identify relationships up to degree eight (e.g. third cousins once removed). Applying the algorithm to all other pairs of cases, we have identified a further 61 putative relative pairs, with an estimated false discovery rate of 10%. The power to identify genealogical links should increase when the new, denser sets of SNP markers are used. Except in populations where there is a searchable electronic database containing virtually all genealogical links in the past six generations, the algorithm should be a useful aid for genealogists working on gene-mapping projects, both linkage studies and association studies. PMID- 15883842 TI - [Malignant paranasal sinus tumors. Diagnosis, therapy and results]. AB - BACKGROUND: Malignant neoplasmas of the paranasal sinuses are rare and present usually in advanced tumor stage due to the lack of early clinical symptoms. PATIENTS AND METHODS: In the last 10 years, 63 patients with paranasal malignancies were treated at the Department of Otolaryngology, Head and Neck Surgery of the University of Wurzburg. 33% of the patients showed an occupational exposition (wood-processing or metal industry). At the time of the first visit to our institution 95% of the patients presented with an extensive disease, staged T3 and T4. Adenocarcinoma (24%), squamous cell carcinoma (22%) and malignant melanoma (19%) were the most common histologies. Surgery combined with radiotherapy was the treatment strategy in 55 patients (87%). RESULTS: Patients with a complete surgical resection showed a higher 5-year-survival rate (77%) than patients with an incomplete resection (56%). In 38% (n=21) of the patients treated with surgery and radiotherapy, a local recurrence of the tumor was observed. This recurrence localised in the skull base and/or the orbita/periorbita occurred most frequently in the first (46%) or the second year (31%). CONCLUSION: The prognosis of malignant paranasal tumors depends mainly on the control of the local tumor growth. Modern strategies of surgical treatment in combination with radiotherapy need to be implemented in an effort to achieve a continuous remission. PMID- 15883843 TI - [Isolated fracture of the handle of malleus. A rare differential diagnosis in cases of conductive hearing loss]. AB - In very rare cases, acute hearing loss can be caused by a fractured long process of the malleus. Such fractures can be due to head traumas, direct injuries of the tympanic membrane or barotraumas. Clinical findings are: abnormal mobility of the manubrium mallei during the Valsalva maneuvre or pneumomassage of the ear drum, conductive hearing loss, abnormal high compliance in the type A tympanigram. We report on a 48 year old female patient who showed typical symptoms and clinical findings, and discuss possible pathogenic factors. In accordance with the literature, we presume that negative pressure in the external ear canal, caused by a rapidly extracted finger (!), might be responsible. At the very onset of hearing loss, the patient noticed a high frequency tinnitus in the affected ear. Surgically, we tried to re-fix the manubrium by splinting it and the adherent parts of the tympanic membrane with cartilage. PMID- 15883844 TI - Role of lipid protrusions in the function of interfacial enzymes. AB - Secretory phospholipase A(2) (sPLA(2)) is a class of interfacially active enzymes that selectively hydrolyze lipid molecules organized at interfaces like membranes. We present a simple theoretical model that relates the sPLA(2) action to the protrusions of the lipid molecules. The model explains (1) the observed enhancement of enzymatic activity by lipids with flexible, neutral, water-soluble polymers linked to their head groups and (2) the lag-burst kinetics of sPLA(2). It yields qualitative predictions of the effect of the initial composition of the membrane, the molecular weight of the polymer, and the composition of the hydrolysis products. PMID- 15883845 TI - [Therapeutic strategies for hereditary retinal diseases]]. PMID- 15883846 TI - [Selective RPE laser treatment with a scanned cw laser beam in rabbits]. AB - BACKGROUND: Selective RPE laser therapy with sparing of the neurosensory layer is possible by applying repetitive microsecond laser pulses. Macular diseases such as diabetic maculopathy, soft confluent drusen due to age-related macular degeneration or central serous chorioretinopathy were shown to be treated successfully-without concurrent laser scotoma-by this technique. It was the goal of this study to show, if selectivity could also be achieved using a conventional green cw-laser by scanning the beam across the retina during irradiation. MATERIAL AND METHODS: A cw-laser beam at 532 nm was coupled to a slitlamp via a single mode optical fiber. The spot (18 microm) was scanned across the retina of Dutch-belted rabbits through a contact lens using a two-dimensional acusto optical deflector. The scan-field was 300 microm x 300 microm in size and consisted of six separate scan lines. The scanning speed was adjusted so as to produce 5 micros exposure at each absorber in the center of the scan line. The entire scan pattern was applied 100 times at each site at a frame rate of 100 Hz. Dose response curve was measured by variation of the laser power. ED(50) thresholds for RPE damage were calculated by fluorescein angiographic leakage in irradiated areas after exposure to different laser intensities. The extent of selectivity was examined by light microscopy. RESULTS: Clinically the selective laser-induced RPE defect was demonstrated by fluorescein angiographic leakage and concurrent absence of ophthalmoscopic visibility. The angiographic ED(50)-damage threshold was 161 mJ/cm(2) (66 mW). Ophthalmoscopic visibility was not noticed even with the maximum available radiant exposure of 438 mJ/cm(2) (180 mW). Thus the safety range between angiographic and ophthalmoscopic thresholds had a factor of at least 2.7. First histological examinations revealed selective RPE destruction with intact photoreceptors for irradiation at laser power levels 2 times above angiographic threshold. CONCLUSION: Selective RPE targeting is feasible with a conventional green cw-laser when scanning the focused laser beam across the fundus with a speed such that every point in exposed RPE is irradiated for duration of 5 micros. PMID- 15883847 TI - Editorial by Prof. Frensch. PMID- 15883848 TI - Intrapopulation variation in gray wolf isotope (delta(15)N and delta(13)C) profiles: implications for the ecology of individuals. AB - Trophic relationships among organisms in terrestrial boreal ecosystems define ecological communities and are important in determining dynamics of energy flow and ecosystem function. We examined trophic relationships between the gray wolf (Canis lupus) and 18 mammalian species from the boreal forest of central Saskatchewan, Canada, using delta(13)C and delta(15)N stable isotope values measured in guard hair samples. Variance in isotope values for wolves and other carnivores was investigated as a proxy for variation in diet among individuals. Isosource, an isotopic source partitioning model, quantified the relative range in proportions of five most-likely prey items in the diets of wolves. The distribution of feasible contributions from each source was dominated by elk (Cervus elaphus; mean: 48%, range:11-75%), followed by white-tailed deer (Odocoileus virginianus; mean: 21%, range: 0-54%), moose (Alces alces; mean:14%, range: 0-41%), beaver (Castor canadensis; mean: 8%, range:0-25%) and snowshoe hare (Lepus americanus; mean: 8%, range: 0-24%). Despite social foraging, our results indicate highly variable diets among individuals and we discuss this in terms of individual versus group ecology of boreal wolves. PMID- 15883849 TI - Contractile forces generated by striae distensae fibroblasts embedded in collagen lattices. AB - Striae distensae are characterized by linear, smooth bands of atrophic-appearing skin that are reddish at first and finally white. They are due to stretching of the skin, as in rapid weight gain, or mechanical stress, as in weight lifting. The pathogenesis of striae distensae is unknown but probably relates to changes in the fibroblast phenotype. In order to characterize striae distensae fibroblasts, alpha-smooth muscle actin expression and contractile forces were studied. Five healthy women with early erythematous striae and five healthy women with older striae were selected. Paired biopsies were taken from the center of lesional striae and adjacent normal skin. Fibroblasts were obtained by an explant technique and expanded in vitro in Dulbecco's modified Eagle's medium. Contractile forces generated by fibroblasts in collagen lattices were measured with the Glasbox device developed in our laboratory. Alpha-smooth muscle actin expression was studied by immunofluorescence labeling of cells and by flow cytometry. Fibroblasts from early striae distensae were the richest cells in alpha-smooth muscle actin filaments and generated the highest contractile forces. Their peak contractile force was 26% greater than normal fibroblasts. There was a 150% higher level of alpha-smooth muscle actin content in fibroblasts from early striae distensae compared with fibroblasts from normal skin. In contrast, there was no significant difference in force generation between old striae fibroblasts and normal fibroblasts with cells expressing no alpha-smooth muscle actin. The contractile properties of fibroblasts from striae distensae varies depending on the stage of the disease. In early striae distensae, fibroblasts acquire a more contractile phenotype, corresponding to that of myofibroblasts. PMID- 15883850 TI - Clinical usefulness of free light chain concentration as a tumor marker in multiple myeloma. AB - Monoclonal immunoglobulin, as a marker for monoclonal gammopathy, is evaluated by protein electrophoresis (PEP) and immunofixation electrophoresis (IFE). However, PEP and IFE are not satisfactory in sensitivity, objectivity, and facility. Recently, a highly sensitive, automated immunoassay for measurement of free light chain (FLC) concentrations in serum and urine has been developed for the identification and monitoring of patients with monoclonal gammopathy. To explore the clinical usefulness of measurement of FLC concentrations, we measured the kappa and lambda FLC concentrations and calculated the kappa/lambda FLC ratios for three groups [multiple myeloma (MM), other diseases, and control] and compared the results of the FLC assay with the results of PEP or IFE. The concentrations of serum kappa and lambda FLCs and the kappa/lambda FLC ratios for the MM group and non-MM groups were distinct. In the MM group, some sera and urine samples had no evidence of M protein on PEP and IFE, but FLC assay showed abnormal concentrations of FLCs and abnormal kappa/lambda FLC ratios in most cases. As compared with the PEP, the kappa/lambda FLC ratio revealed higher sensitivity in all diagnostic ranges with different cutoff values. Particularly, when the cutoff value 2.0 for kappa/lambda FLC ratio was used, specificity and positive predictive value were largely improved than when the cutoff values 1.2 and 1.5 were used. These findings indicated that FLC assay enables to detect myeloma patients with very low M protein due to early stage or after therapy and to distinguish patients with monoclonal increase of FLC from patients with polyclonal increase of FLC due to other conditions, particularly using kappa/lambda FLC ratio 0.3-2.0 as a diagnostic range. Despite some technical limitations of the assay, the incorporation of kappa/lambda FLC ratios with FLC concentrations is useful in the detection of M protein, particularly with negative serum or urine IFE results, and differentiation of monoclonal gammopathies from patients with polyclonal increase in FLC due to other conditions. PMID- 15883851 TI - Anatomy, histology, histochemistry and fine structure of the Harderian gland in the South American armadillo Chaetophractus villosus (Xenarthra, Mammalia). AB - The anatomical, histological, histochemical and ultrastructural characteristics of the Harderian gland of the armadillo Chaetophractus villosus were described. The gland is the largest structure in the bony orbit. It is situated in the anteroventral region of the orbit. Obvious structural differences are not observed between males and females. The gland is compound-branched tubulo alveolar, being characterized by a single layer of columnar cells surrounded by myoepithelial cells. It possesses a single excretory duct opened into the inner canthus. All glandular cells show yellow-green autofluorescence and additionally some glandular lumen may contain dense autofluorescent solid accretions. There are two peculiar and outstanding cytoplasmic features. One is represented by the smooth endoplasmic reticulum (SER), forming a closely woven meshwork. The other one is represented by "membranous bodies" apparently derived from the SER, RER and cytoskeleton with a "Star of David" configuration situated in the supranuclear region. Three types of vesicles are detected in the cytoplasm. Histochemical staining methods reveal lipids, proteins, neutral and acidic containing glycoconjugates in secretory vesicles. The mechanism of secretion appears either merocrine or apocrine. The epithelium of the intra- and inter lobular excretory ducts suggests secretory activity. Tubulo-acinar glands similar to those seen in the lacrimal gland and nictitans glands are found related to the intralobular and main excretory ducts. The capillary network is characterized by fenestrated endothelium. The stroma possesses unmyelinated axons and plasma cells. The normal secretion of the secretory endpieces, particularly lipids, proteins and glycoconjugates, is complemented by mucous and serous secretions released by ductal cells and glands associated to the ducts. PMID- 15883852 TI - Mycorrhizal inoculum potentials of pure reclamation materials and revegetated tailing sands from the Canadian oil sand industry. AB - Recent improvements in the management of oil sand tailings used by the Canadian oil sand industry have resulted in the production of composite tailing sands (CT): a new challenging material for reclamation work. Jack pine (Pinus banksiana Lamb.), hybrid poplar (Populus deltoides Bartr. ex Marsh. xPopulus nigra L.) and red clover (Trifolium pratense L.) plants were used in an 8-week greenhouse bioassay to evaluate the mycorrhizal inoculum potential of CT. This inoculum potential was compared with that of three other reclamation materials [common tailing sands (TS), deep overburden (OB) and muskeg peat (MK)], and with three sites reclaimed in 1982 (R82), 1988 (R88) and 1999 (R99). CT was devoid of active mycorrhizal propagules while all other materials showed some level of inoculum potential. Arbuscular mycorrhizal fungi were observed on roots of clover or poplar grown in TS, OB, and all substrates containing peat (MK, R82, R88 and R99). Pine roots were also colonized by vesicle-forming hyphae of an unidentified fine endophyte and by dark septate fungi. Ectomycorrhizas (ECM) were observed on pine and poplar grown in OB, MK, and in soils from the two older reclaimed sites (R82 and R88). Using morpho- and molecular typing, six ECM fungi were identified to the genus or species level: Laccaria sp., Thelephora americana, Wilcoxina sp. (E-strain), Tuber sp. (I-type), a Sebacinoid, and a Pezizales species. Laccaria sp. and Wilcoxina sp. were the most frequently observed ECM species. PMID- 15883853 TI - Mycorrhizal C costs and nutritional benefits in developing grapevines. AB - Arbuscular mycorrhizal (AM) C-costs in grapevines were investigated. Dormant vines rely on stored C for initial growth. Therefore AM colonisation costs would compete with plant growth for available C reserves. One-year-old grapevines, colonised with Glomus etunicatum (Becker and Gerdemann), were cultivated under glasshouse conditions. The C-economy and P utilisation of the symbiosis were sequentially analysed. AM colonisation, during the 0-67 day growth period, used more stem C relative to root C, which resulted in lower shoot growth. The decline in AM colonisation during the period of 67-119 days coincided with stem C replenishment and higher shoot growth. Construction costs of AM plants and root C allocation increased with root P uptake. The efficiency of P utilisation was lower in AM roots. The reliance of AM colonisation on stem C declined with a decrease in colonisation, providing more C for the refilling of stem carbohydrate reserves and shoot growth. Once established, the AM symbiosis increased P uptake at the expense of refilling of root C reserves. Although higher root C allocation increased plant construction costs, AM roots were more efficient at P utilisation. PMID- 15883854 TI - Ethnic differences in allele frequency of autoimmune-disease-associated SNPs. AB - Several multiple, large-scale, genetic studies on autoimmune-disease-associated SNPs have been reported recently: peptidylarginine deiminase type 4 (PADI4) in rheumatoid arthritis (RA); solute carrier family 22 members 4 and 5 (SLC22A4 and 5) in RA and Crohn's disease (CD); programmed cell death 1 (PDCD1) in systemic lupus erythematosus (SLE), type 1 diabetes mellitus (T1D), and RA; and protein tyrosine phosphatase nonreceptor type 22 (PTPN22) in T1D, RA, and SLE. Because these reports on association were not always evaluated in multiple ethnic groups and because ethnic difference in allele frequency of the variants has been also reported, we investigated allele frequencies of nine SNPs in four autoimmune disease-associated loci in Caucasian, African-descent, and Japanese populations. Although SNPs in PADI4 had similar allele frequency among three groups [maximal difference 11%; (P >0.05)], the other three loci revealed statistically significant allele frequency differences (maximal difference 39% (P <0.00001), 13% (P <0.00001), and 8% (P <0.00001) in SLC22A4, PDCD1, and PTPN22, respectively). Of note, three SNPs in the three loci that had allele frequency more than 8% in the Caucasian population were either not polymorphic at all or extremely rare in the Japanese population. Our data suggest that ethnic variations of polymorphisms should be evaluated in detail, and differences should be incorporated into investigations of susceptibility variants for common diseases. PMID- 15883855 TI - Size of the protein-coding genome and rate of molecular evolution. AB - In diploid populations of size N, there will be 2 Nmu mutations per nucleotide (nt) site (or per locus) per generation (mu stands for mutation rate). If either the population or the coding genome double in size, one expects 4 Nmu mutations. What is important is not the population size per se but the number of genes (coding sites), the two being often interconverted. Here we compared the total physical length of protein-coding genomes (n) with the corresponding absolute rates of synonymous substitution (K(S)), an empirical neutral reference. In the classical occupancy problem and in the coupons collector (CC) problem, n was expressed as the mean rate of change (K(CC)). Despite inherently very low power of the approaches involving averaging of rates, the mode of molecular evolution of the total size phenotype of the coding genome could be evidenced through differences between the genomic estimates of K(CC) [K(CC)=1/(ln n + 0.57721) n] and rate of molecular evolution, K(S). We found that (1) the estimates of n and K(S) are reciprocally correlated across taxa (r=0.812; p<< 0.001); (2) the gamete cell division hypothesis (Chang et al. Proc Natl Acad Sci USA 91:827-831, 1994) can be confirmed independently in terms of K(CC)/K(S) ratios; (3) the time scale of molecular evolution changes with change in mutation rate, as previously shown by Takahata (Proc Natl Acad Sci USA 87:2419-2423, 1990), Takahata et al. (Genetics 130:925-938, 1992), and Vekemans and Slatkin (Genetics 137:1157-1165, 1994); (4) the generation time and population size (Lynch and Conery, Science 302:1401-1404, 2003) effects left their "signatures" at the level of the size phenotype of the protein-coding genome. PMID- 15883856 TI - Pessary use in advanced pelvic organ prolapse. AB - The objective of this study was to review our experience with pessary use for advanced pelvic organ prolapse. Charts of patients treated for Stage III and IV prolapse were reviewed. Comparisons were made between patients who tried or refused pessary use. A successful trial of pessary was defined by continued use; a failed trial was defined by a patient's discontinued use. Thirty-two patients tried a pessary; 45 refused. Patients who refused a pessary were younger, had lesser degree of prolapse, and more often had urinary incontinence. Most patients (62.5%) continued pessary use and avoided surgery. Unsuccessful trial of pessary resorting to surgery included four patients (33%) with unwillingness to maintain, three patients (25%) with inability to retain and two patients (17%) with vaginal erosion and/or discharge. Our findings suggest that pessary use is an acceptable first-line option for treatment of advanced pelvic organ prolapse. PMID- 15883858 TI - International Consultation on IC - Rome, September 2004/Forging an International Consensus: progress in painful bladder syndrome/interstitial cystitis. Report and abstracts. PMID- 15883859 TI - Uterine fibroid embolization: more evidence is required. PMID- 15883860 TI - Modified PAIR technique for percutaneous treatment of high-risk hydatid cysts. AB - PURPOSE: This paper presents a modification of the known method for percutaneous treatment of hydatid cyst, the PAIR technique. It aimed to achieve safe aspiration of large symptomatic cysts and cysts with a danger of impending rupture. METHODS: We designed a coaxial catheter system to achieve concomitant evacuation of cyst contents while infusing scolicidal agent. Hypertonic saline is used to wash out cyst contents and to kill protoscolices. This was followed by injection of a sclerosant (ethyl alcohol 95%) into the residual cyst cavity to prevent formation of a cyst collection after the procedure. Seventeen cysts in 14 patients were successfully aspirated. Follow-up plain radiographs, ultrasonography and CT were performed weekly in the first 4 weeks and then at 3, 6 and 12 months for all patients. Seven patients (9 drained cysts) were followed up for 2 years and 1 patient for 3 years. RESULTS: All cysts were successfully aspirated. The following morphologic changes were noticed: a gradual decrease in cyst size (17 cysts, 100%), thickening and irregularity of the cyst wall due to separation of endocyst from pericyst (7 cysts, 41%), development of a heterogeneous appearance of the cyst components (8 cysts, 47%) and development of pseudotumor (2 cysts, 12%). None of the treated cysts disappeared completely. No significant procedure-related complications were encountered. CONCLUSION: This modified PAIR technique is a reliable method for percutaneous treatment of risky and symptomatic hydatid cysts. PMID- 15883861 TI - Percutaneous venous thrombectomy using the Arrow-Trerotola percutaneous thrombolytic device (PTD) with temporary caval filtration: in vitro investigations. AB - PURPOSE: To evaluate the size and quantity of downstream emboli after thrombectomy using the Arrow-Trerotola Percutaneous Thrombolytic Device (PTD) with or without temporary filtration for extensive iliofemoral and iliocaval thrombi in an in vitro flow model. METHODS: Iliocaval thrombi were simulated by clotted bovine blood in a flow model (semilucent silicone tubings, diameter 12-16 mm). Five experimental set-ups were performed 10 times each; thrombus particles and distribution were measured in the effluent. First, after retrograde insertion, mechanical thrombectomy was performed using the PTD alone. Then a modified self-expanding tulip-shaped temporary vena cava stent filter was inserted additionally at the beginning of each declotting procedure and removed immediately after the intervention without any manipulation within or at the filter itself. In a third step, the filter was filled with thrombus only. Here, two experiments were performed: Careful closure within the flow circuit without any additional fragmentation procedure and running the PTD within the filter lumen, respectively. In the final set-up, mechanical thrombectomy was performed within the thrombus-filled tubing as well as in the filter lumen. The latter was closed at the end of the procedure and both devices were removed from the flow circuit. RESULTS: Running the PTD in the flow circuit without filter protection led to a fragmentation of 67.9% (+/-7.14%) of the clot into particles < or =500 microm; restoration of flow was established in all cases. Additional placement of the filter safely allowed maceration of 82.9% (+/-5.59%) of the thrombus. Controlled closure of the thrombus-filled filter within the flow circuit without additional mechanical treatment broke up 75.2% (+/-10.49%), while additional mechanical thrombectomy by running the PTD within the occluded filter led to dissolution of 90.4% (+/-3.99%) of the initial clot. In the final set-up, an overall fragmentation rate of 99.6% (+/-0.44%) was achieved. CONCLUSIONS: The combined use of the Arrow-Trerotola PTD and a temporary vena cava stent filter proved to be effective for even large clot removal in this experimental set-up. PMID- 15883862 TI - Endovascular treatment of persistent epistaxis due to pseudoaneurysm formation of the ophthalmic artery secondary to nasogastric tube. AB - We present the case of a 60-year-old man with persistent epistaxis for 20 days that had started 2 weeks after removal of a nasogastric tube placed for an abdominal operation. There was no pathologic finding at selective facial and internal maxillary artery injections. An injury to the ethmoidal branches of the ophthalmic arteries or other arterial origins of bleeding was suspected. The internal carotid artery angiography revealed a pseudoaneurysm of an anterior ethmoidal branch of the left ophthalmic artery. The pseudoaneurysm was occluded with NBCA-histoacryl (25%) injection. PMID- 15883863 TI - Cyanobacteria and biodeterioration of cultural heritage: a review. AB - Growing concern for the preservation of cultural heritage has led to a greater interest in the biological attack on these buildings. The importance of cyanobacteria as deteriogens is emphasized and the traditional and more modern molecular methods used to detect these microorganisms are discussed. The development of molecular techniques for the rapid identification of cyanobacteria without need for culture and isolation is fundamental if our knowledge of these communities in biofilms on the surfaces of historic buildings is to be extended. PMID- 15883864 TI - Microbial diversity during biodegradation of crude oil in seawater from the North Sea. AB - Microbial communities were characterized during biodegradation of immobilized oil in seawater from the Statfjord field and the German Bight in the North Sea. Seawater samples were collected at different distances from pollution sources at the two locations. A Statfjord oil was immobilized on hydrophobic synthetic Fluortex fabrics and submerged in closed flasks (no headspace) with natural or sterile seawater and incubated at 13 degrees C for 56 days. Biodegradation of immobilized n-alkanes was measured by gas chromatography, total microbes were enumerated by epifluorescence microscopy, and culturable heterotrophic and oil degrading microorganisms were quantified by most probable number (MPN) analysis. Polymerase chain reaction (PCR) amplification of bacterial 16S rDNA in water samples was conducted during biodegradation experiments. The amplified 16S rDNA fragments were characterized by denaturing gradient gel electrophoresis (DGGE), and by sequence analysis of cloned inserts. Biodegradation rates of alkanes in seawater collected at different distances from the pollution sources did not differ significantly (P > 0.05). Concentrations of oil-degrading microorganisms showed a temporary peak after 7 days of degradation, with a subsequent decline later in the period. DGGE analysis of 16S rRNA genes showed that community diversity decreased during the first 2-3 weeks of biodegradation, with the emergence of a few dominant bands. Cloning, restriction analysis, and sequence analysis of the 16S rDNA fragments revealed >30 different phylotypes. Abundant types during biodegradation belonged to the alpha-Proteobacteria, in waters from both Statfjord and the German Bight. Cloning and sequencing studies indicated that the most abundant bacteria during biodegradation belonged to the family Rhodobacteraceae, with the closest relationship to the genera Sulfitobacter and Roseobacter. PMID- 15883865 TI - The role of pigmentation, ultraviolet radiation tolerance, and leaf colonization strategies in the epiphytic survival of phyllosphere bacteria. AB - Phenotypic mechanisms that enhance bacterial UVR survival typically include pigmentation and DNA repair mechanisms which provide protection from UVA and UVB wavelengths, respectively. In this study, we examined the contribution of pigmentation to field survival in Clavibacter michiganensis and evaluated differences in population dynamics and leaf colonization strategies. Two C. michiganensis pigment-deficient mutants were significantly reduced in UVA radiation survival in vitro; one of these mutants also exhibited reduced field populations on peanut when compared to the wild-type strain over the course of replicate 25-day experiments. The UVR-tolerant C. michiganensis strains G7.1 and G11.1 maintained larger epiphytic field populations on peanut compared to the UVR sensitive C. michiganensis T5.1. Epiphytic field populations of C. michiganensis utilized the strategy of solar UVR avoidance during leaf colonization resulting in increased strain survival on leaves after UVC irradiation. These results further demonstrate the importance of UVR tolerance in the ability of bacterial strains to maintain population size in the phyllosphere. However, an examination of several bacterial species from the peanut phyllosphere and a collection of environmental Pseudomonas spp. revealed that sensitivity to UVA and UVC radiation was correlated in some but not all of these bacteria. These results underscore a need to further understand the biological effects of different solar wavelength groups on microbial ecology. PMID- 15883866 TI - Molecular analysis of bacterial community based on 16S rDNA and functional genes in activated sludge enriched with 2,4-dichlorophenoxyacetic acid (2,4-D) under different cultural conditions. AB - Differential emergence and diversity of bacterial communities from activated sludge in response to varied cultural conditions using 2,4-dichlorophenoxyacetic acid (2,4-D) were investigated by coupling molecular analyses based on 16S rDNA with functional genes. We employed three different cultural conditions: (1) a culture sequentially fed a high concentration (300 mg/L) of 2,4-D (HS); (2) a culture continuously fed a low concentration (10 mg/L) of 2,4-D (LC); and (3) a serial batch culture in which 1% (v/v) of culture was transferred to a fresh medium containing a high concentration (300 mg/L) of 2,4-D (HB). The HS and LC bioreactors were operated for 3 months and HB was repeatedly transferred for 1 month. The 2,4-D was stably degraded under all the cultural conditions tested. PCR amplification and cloning-based analysis of functional genes using community DNAs from the cultures revealed five different oxygenase genes that may be involved in the initial step of 2,4-D degradation. All five gene-types were present in HS, while one of the five genes, type V (tftA) was not detected in LC. Quantitative PCR analysis showed that in HS, Ralstonia eutropha JMP 134 type tfdA4 (type I) was the most abundant in copy number (2.0 +/- 0.1 x 10(7) copies/microg DNA) followed by RASC type-tfdA (type II) (1.8 +/- 1.0 x 10(6) copies/microg DNA), putative cadA-like gene (type IV) (2.6 +/- 0.8 x 10(5) copies/microg DNA), cadA gene (type III) (1.3 +/- 1.0 x 10(4) copies/microg DNA), and tftA gene (type V) (3.5 +/- 1.1 x 10(3) copies/microg DNA). Similar results were obtained in LC. In contrast, HB contained only type I and type III genes, and the type I gene was five orders of magnitude greater in copy number than the type III gene. Denaturing gel gradient electrophoresis (DGGE) analysis of PCR, amplified 16S rDNA fragments of bacterial communities in the three different cultures showed low similarity coefficient values (< or =0.35) when compared to the original activated sludge, suggesting that 2,4-D amendment caused a drastic change in the bacterial community. Particularly, HB showed only six bands (16-18 bands in the other cultures) and very low similarity coefficient values when compared to the other communities (0.10 to HS, 0.17 to LC, and 0.0 to original sludge). These results indicated that serial batch culturing (HB) resulted in a phylogenetically limited number of 2,4-D degrading bacteria carrying limited catabolic genes whereas more diverse 2,4-D degraders and catabolic genes were present in HS and LC. Therefore, the approach used for monitoring should be taken into account when one evaluates the population dynamics of contaminant-degrading bacteria at bioremediation sites. PMID- 15883867 TI - Cloning and expression of fructosyl-amine oxidase from marine yeast Pichia species N1-1. AB - The gene encoding the fructosyl-amine oxidase (FAOD) from the marine yeast Pichia sp. N1-1 was cloned and expressed in Escherichia coli. Partial amino acid sequence analysis of the Pichia sp. N1-1 FAOD allowed the design of oligonucleotide primers for the amplification of the gene by inverse polymerase chain reaction. The FAOD gene was found to be devoid of introns and to encode a 48-kDa protein composed of 429 amino acid residues. The FAD-binding consensus sequence GXGXXG: and the FAD covalent attachment-site cysteine residue have been identified within the predicted amino acid sequence. Comparisons with the amino acid sequences of other eukaryotic FAODs showed only 30% to 40% identities, establishing that the isolated Pichia N1-1 gene encodes a unique FAOD. Recombinant FAOD expression levels in E. coli reached 0.48 U/mg of soluble protein, which is considerably greater than native expression levels by inducing Pichia sp. N1-1 with fructosyl-valine (f-Val). The kinetic properties of the recombinant enzyme were almost indistinguishable from those of the native enzyme. We previously reported on the construction of a number of effective Pichia sp. N1 1 FAOD-based biosensors for measuring f-Val, a model compound for glycated hemoglobin. The further development of these biosensor systems can now greatly benefit from protein engineering and recombinant expression of the FAOD from Pichia N1-1. PMID- 15883868 TI - Topographic, bioclimatic, and vegetation characteristics of three ecoregion classification systems in North America: comparisons along continent-wide transects. AB - Ecoregion classification systems are increasingly used for policy and management decisions, particularly among conservation and natural resource managers. A number of ecoregion classification systems are currently available, with each system defining ecoregions using different classification methods and different types of data. As a result, each classification system describes a unique set of ecoregions. To help potential users choose the most appropriate ecoregion system for their particular application, we used three latitudinal transects across North America to compare the boundaries and environmental characteristics of three ecoregion classification systems [Kuchler, World Wildlife Fund (WWF), and Bailey]. A variety of variables were used to evaluate the three systems, including woody plant species richness, normalized difference in vegetation index (NDVI), and bioclimatic variables (e.g., mean temperature of the coldest month) along each transect. Our results are dominated by geographic patterns in temperature, which are generally aligned north-south, and in moisture, which are generally aligned east-west. In the west, the dramatic changes in physiography, climate, and vegetation impose stronger controls on ecoregion boundaries than in the east. The Kuchler system has the greatest number of ecoregions on all three transects, but does not necessarily have the highest degree of internal consistency within its ecoregions with regard to the bioclimatic and species richness data. In general, the WWF system appears to track climatic and floristic variables the best of the three systems, but not in all regions on all transects. PMID- 15883869 TI - Identifying ecoregion boundaries. AB - This article summarizes the rationale I used in identifying ecoregion boundaries on maps of the United States, North America, and the world's continents, published from 1976 to 1998. The geographic reasoning used in drawing boundaries involves 20 principles, which are presented to stimulate discussion and further understanding. Brief background and references are provided for the principles. PMID- 15883870 TI - Potential of multivariate quantitative methods for delineation and visualization of ecoregions. AB - Multivariate clustering based on fine spatial resolution maps of elevation, temperature, precipitation, soil characteristics, and solar inputs has been used at several specified levels of division to produce a spectrum of quantitative ecoregion maps for the conterminous United States. The coarse ecoregion divisions accurately capture intuitively-understood regional environmental differences, whereas the finer divisions highlight local condition gradients, ecotones, and clines. Such statistically generated ecoregions can be produced based on user selected continuous variables, allowing customized regions to be delineated for any specific problem. By creating an objective ecoregion classification, the ecoregion concept is removed from the limitations of human subjectivity, making possible a new array of ecologically useful derivative products. A red-green-blue visualization based on principal components analysis of ecoregion centroids indicates with color the relative combination of environmental conditions found within each ecoregion. Multiple geographic areas can be classified into a single common set of quantitative ecoregions to provide a basis for comparison, or maps of a single area through time can be classified to portray climatic or environmental changes geographically in terms of current conditions. Quantified representativeness can characterize borders between ecoregions as gradual, sharp, or of changing character along their length. Similarity of any ecoregion to all other ecoregions can be quantified and displayed as a "representativeness" map. The representativeness of an existing spatial array of sample locations or study sites can be mapped relative to a set of quantitative ecoregions, suggesting locations for additional samples or sites. In addition, the shape of Hutchinsonian niches in environment space can be defined if a multivariate range map of species occurrence is available. PMID- 15883871 TI - Characterization of a 4 kb variant of the nifD element in Anabaena sp. strain ATCC 33047. AB - Heterocyst differentiation in some cyanobacteria is accompanied by a programmed DNA rearrangement within the nitrogen fixation gene nifD. The nifD element is excised from within nifD during the latter stages of heterocyst differentiation by site-specific recombination. There is considerable variation in those nifD elements examined thus far, with Nostoc sp. Strain PCC 7120 and Anabaena variabilis having 11 kb elements, and Nostoc punctiforme having a 24 kb element. Here we characterize a 4 kb nifD element in Anabaena sp. Strain ATCC 33047, and compare it with the other sequenced nifD elements. While there is considerable variation in both the size (ranging from 4 kb to 24 kb) and composition of the nifD elements examined thus far, there are regions that are conserved in all. These conserved regions include the flanking 3' and 5' regions, the xisA gene, and a small open reading frame known as ORF2 in Nostoc sp. Strain PCC 7120. PMID- 15883872 TI - Involvement of hexokinase Hxk1 in glucose catabolite repression of LIP2 encoding extracellular lipase in the yeast Yarrowia lipolytica. AB - The yeast Yarrowia lipolytica produces an extracellular lipase encoded by the LIP2 gene. However, very little is known about the mechanisms controlling its expression, especially on glucose media. In this work, the involvement of hexokinase Hxk1 in the glucose catabolite repression of LIP2 was investigated in a lipase overproducing mutant less sensitive to glucose repression. This mutant has a reduced capacity to phosphorylate hexose compared with the wild-type strain, but no differences could be observed between the HXK1 sequences in the two isolates. This suggested that the reduced phosphorylating activity of the mutant strain probably resulted from a modification in the level of HXK1 expression. However, overexpression of the HXK1 gene in this mutant led to a decrease of both LIP2 induction and extracellular lipase activity, suggesting that the hexokinase is involved in the glucose catabolite repression of LIP2 in Y lipolytica. PMID- 15883873 TI - Removing selenite from groundwater with an in situ biobarrier: laboratory studies. AB - Laboratory biobarriers were evaluated for their ability to remove selenite from flowing groundwater. Microbial activity in aquifers is usually limited by substrate availability, and biobarriers stimulate microbial activity by providing a substrate; for these studies soybean oil was used. Water containing 10 mg L(-1) selenite-Se was pumped through the biobarriers for 74 days and the amount present in the effluent monitored. The amounts remained high for the first 2 weeks of the study but then declined. From day 28 until the end of the study the amount of selenite-Se in the column effluents averaged 0.20 +/- 0.04 mg L(-1), a decrease of approximately 98%. At the end of the study about half of the selenite-Se applied to the columns was recovered as immobilized selenium trapped by the biobarrier. This study suggests that biobarriers containing vegetable oil might be used as a process for removing selenite from contaminated groundwater. PMID- 15883874 TI - Transcriptional regulation of beta-glucanase activity in the ruminal bacterium, Prevotella bryantii B14. AB - The ruminal bacterium, Prevotella bryantii B(1)4, grew more rapidly with glucose as an energy source than mannose (0.73 versus 0.47 h(-1)) and had 8-fold less beta-glucanase activity (50 versus 400 nmol reducing sugar mg protein(-1) min( 1)). Cultures that were provided with glucose and mannose had little beta glucanase activity even though both sugars were utilized simultaneously. The observation that glucose and mannose were utilized simultaneously indicated that beta-glucanase expression was not merely a simple induction or inducer exclusion. When glucose was added to cultures growing on mannose, hexose flux through the glucomannokinase increased 1.5-fold, and this increase was associated with an almost immediate decrease in beta-glucanase mRNA. After only three generation (doubling) times, the amount of beta-glucanase mRNA was comparable to that observed in cells growing only with glucose. These results indicate that beta glucanase activity is transcriptionally regulated. However, further work will be needed to define more precisely the nature of this regulation and to identify the intermediate in this response. PMID- 15883875 TI - Cytokine response of lymphocytes persistently infected with Chlamydia pneumoniae. AB - Chlamydia pneumoniae infection of lymphocytes in blood has been documented, and it is apparent that control of this pathogen in lymphocytes as well as immune functions of the infected lymphocytes may be critical in the development of chronic inflammatory diseases associated with infection by this bacterium. Since immune function of lymphocytes infected with C. pneumoniae has not been well studied, the cytokine response of lymphocytes infected with this pathogen was analyzed using an in vitro infection model of the Molt-4 human lymphoid cell line. C. pneumoniae infection of the cells showed a persistent infection without any vigorous growth of the bacteria. Analysis of the cytokine response of the cells persistently infected with C. pneumoniae showed minimum induction of inflammatory cytokine TNF-alpha message, determined by real-time reverse transcription (RT)-PCR in the lymphocytes, even though the infection of THP-1 monocytic cells showed a marked induction of this cytokine messages. BIC (a lymphocyte activation marker gene) as well as IFN-gamma messages were also minimally induced by the infection in Molt-4 lymphocytes. In contrast, constitutive expression of interleukin 8 (IL-8) messages of Molt-4 cells was suppressed by the infection. Thus, these results suggest that lymphocytes persistently infected with C. pneumoniae may have attenuated cytokine responses. PMID- 15883876 TI - Differential rhizosphere establishment and cyanide production by alginate formulated weed-deleterious rhizobacteria. AB - The effects of Pseudomonas putida ATH2-1RI/9 and Acidovorax delafieldii ATH2 2RS/1 on rhizosphere colonization, cyanide production, and growth of velvetleaf and corn was examined. When formulated in alginate beads and inoculated onto velvetleaf and corn plants (10(9) CFU/plant), only P. putida ATH2-1RI/9 consistently reduced velvetleaf growth. Neither isolate inhibited corn growth. Interestingly the levels of P. putida ATH2-1RI/9 in the velvetleaf rhizosphere were 1000-fold higher (7 x 10(7) CFU/g root) than the A. delafieldii ATH2-2RS/1 populations. Cyanide (53-68 mM/g root) was recovered from the P. putida ATH2 1RI/9-inoculated velvetleaf plants. In contrast both A. delafieldii ATH2-2RS/1 and P. putida ATH2-1RI/9 colonized the corn rhizosphere to the same extent (1-5 x 10(7) CFU/g root), producing 1 mM and 14 mM/g root respectively. These results suggest that bacterial formulation methods can influence the effectiveness of deleterious rhizobacteria in reducing weed growth. PMID- 15883877 TI - Compositional properties of green-plant plastid genomes. AB - We studied variation of GC contents among plastid (Pt) genomes of green plants. In the green plants, the GC contents of the whole Pt genomes range from 42.14 to 28.81%. These values are similar to those observed in the mitochondrial (Mt) genomes of the green plants, however, the GC contents in the Pt genomes are not related to those in the Mt genomes or the nuclear (Nc) genomes. In addition, some compositional properties of the three types of genomes are different. Thus, it is suggested that the GC contents of the Pt genomes are maintained independently of the other genomes within a cell. We found that the compositional bias toward AT is strong at the third codon position and in intergenic spacer (IGS) regions in the Pt genomes, and the GC contents (GC3 and GCIGS) at these sites are generally similar within each genome. Additionally, the GC3 and GCIGS are strongly related to the whole-genome GC content. Therefore, the interspecific variation of the GC contents in the Pt genomes is suggested to be mainly caused by the variation of the GC3 and GCIGS, both of which are considered to be under weak selective constraints. Using a maximum likelihood approach, we estimated equilibrium GC3 (eqGC(3)) of 12 genes in the land-plant Pt genomes. We found an increase in eqGC(3) after the divergence of liverworts. These results suggest that genome wide factors such as GC mutational bias are important for the biased base composition in the Pt genomes. PMID- 15883878 TI - Accuracy and power of the likelihood ratio test for comparing evolutionary rates among genes. AB - Sequences for multiple protein-coding genes are now commonly available from several, often closely related species. These data sets offer intriguing opportunities to test hypotheses regarding whether different types of genes evolve under different selective pressures. Although maximum likelihood (ML) models of codon substitution that are suitable for such analyses have been developed, little is known about the statistical properties of these tests. We use a previously developed fixed-sites model and computer simulations to examine the accuracy and power of the likelihood ratio test (LRT) in comparing the nonsynonymous-to-synonymous substitution rate ratio (omega = dN/dS) between two genes. Our results show that the LRT applied to fixed-sites models may be inaccurate in some cases when setting significance thresholds using a chi(2) approximation. Instead, we use a parametric bootstrap to describe the distribution of the LRT statistic for fixed-sites models and examine the power of the test as a function of sampling variables and properties of the genes under study. We find that the power of the test is high (>80%) even when sampling few taxa (e.g., six species) if sequences are sufficiently diverged and the test is largely unaffected by the tree topology used to simulate data. Our simulations show fixed-sites models are suitable for comparing substitution parameters among genes evolving under even strong evolutionary constraint (omega approximately 0.05), although relative rate differences of 25% or less may be difficult to detect. PMID- 15883879 TI - Divergent histories of rDNA group I introns in the lichen family Physciaceae. AB - The wide but sporadic distribution of group I introns in protists, plants, and fungi, as well as in eubacteria, likely resulted from extensive lateral transfer followed by differential loss. The extent of horizontal transfer of group I introns can potentially be determined by examining closely related species or genera. We used a phylogenetic approach with a large data set (including 62 novel large subunit [LSU] rRNA group I introns) to study intron movement within the monophyletic lichen family Physciaceae. Our results show five cases of horizontal transfer into homologous sites between species but do not support transposition into ectopic sites. This is in contrast to previous work with Physciaceae small subunit (SSU) rDNA group I introns where strong support was found for multiple ectopic transpositions. This difference in the apparent number of ectopic intron movements between SSU and LSU rDNA genes may in part be explained by a larger number of positions in the SSU rRNA, which can support the insertion and/or retention of group I introns. In contrast, we suggest that the LSU rRNA may have fewer acceptable positions and therefore intron spread is limited in this gene. PMID- 15883880 TI - Phylogenetic differences in content and intensity of periodic proteins. AB - Many proteins exhibit sequence periodicity, often correlated with a visible structural periodicity. The statistical significance of such periodicity can be assessed by means of a chi-squared-based test, with significance thresholds being calculated from shuffled sequences. Comparison of the complete proteomes of 45 species reveals striking differences in the proportion of periodic proteins and the intensity of the most significant periodicities. Eukaryotes tend to have a higher proportion of periodic proteins than eubacteria, which in turn tend to have more than archaea. The intensity of periodicity in the most periodic proteins is also greatest in eukaryotes. By contrast, the relatively small group of periodic proteins in archaea also tend to be weakly periodic compared to those of eukaryotes and eubacteria. Exceptions to this general rule are found in those prokaryotes with multicellular life-cycle phases, e.g., Methanosarcina sp., or Anabaena sp., which have more periodicities than prokaryotes in general, and in unicellular eukaryotes, which have fewer than multicellular eukaryotes. The distribution of significantly periodic proteins in eukaryotes is over a wide range of period lengths, whereas prokaryotic proteins typically have a more limited set of period lengths. This is further investigated by repeating the analysis on the NRL-3D database of proteins of solved structure. Some short-range periodicities are explicable in terms of basic secondary structure, e.g., alpha helices, while middle-range periodicities are frequently found to consist of known short Pfam domains, e.g., leucine-rich repeats, tetratricopeptides or armadillo domains. However, not all can be explained in this way. PMID- 15883881 TI - Comparison of the PhoPQ regulon in Escherichia coli and Salmonella typhimurium. AB - The PhoPQ two-component system acts as a transcriptional regulator that responds to Mg(2+) starvation both in Escherichia coli and Salmonella typhimurium (Garcia et al. 1996; Kato et al. 1999). By monitoring the availability of extracellular Mg(2+), this two-component system allows S. typhimurium to sense the transition from an extracellular environment to a subcellular location. Concomitantly with this transition, a set of virulence factors essential for survival in the intracellular environment is activated by the PhoPQ system (Groisman et al. 1989; Miller et al. 1989). Compared to nonpathogenic strains, such as E. coli K12, the PhoPQ regulon in pathogens must contain target genes specifically contributing to the virulence phenotype. To verify this hypothesis, we compared the composition of the PhoPQ regulon between E. coli and S. typhimurium using a combination of expression experiments and motif data. PhoPQ-dependent genes in both organisms were identified from PhoPQ-related microarray experiments. To distinguish between direct and indirect targets, we searched for the presence of the regulatory motif in the promoter region of the identified PhoPQ-dependent genes. This allowed us to reconstruct the direct PhoPQ-dependent regulons in E. coli K12 and S. typhimurium LT2. Comparison of both regulons revealed a very limited overlap of PhoPQ-dependent genes between both organisms. These results suggest that the PhoPQ system has acquired a specialized function during evolution in each of these closely related species that allows adaptation to the specificities of their lifestyles (e.g., pathogenesis in S. typhimurium). PMID- 15883882 TI - Comparison of the yeast proteome to other fungal genomes to find core fungal genes. AB - The purpose of this research was to search for evolutionarily conserved fungal sequences to test the hypothesis that fungi have a set of core genes that are not found in other organisms, as these genes may indicate what makes fungi different from other organisms. By comparing 6355 predicted or known yeast (Saccharomyces cerevisiae) genes to the genomes of 13 other fungi using Standalone TBLASTN at an e-value <1E-5, a list of 3340 yeast genes was obtained with homologs present in at least 12 of 14 fungal genomes. By comparing these common fungal genes to complete genomes of animals (Fugu rubripes, Caenorhabditis elegans), plants (Arabidopsis thaliana, Oryza sativa), and bacteria (Agrobacterium tumefaciens, Xylella fastidiosa), a list of common fungal genes with homologs in these plants, animals, and bacteria was produced (938 genes), as well as a list of exclusively fungal genes without homologs in these other genomes (60 genes). To ensure that the 60 genes were exclusively fungal, these were compared using TBLASTN to the major sequence databases at GenBank: NR (nonredundant), EST (expressed sequence tags), GSS (genome survey sequences), and HTGS (unfinished high-throughput genome sequences). This resulted in 17 yeast genes with homologs in other fungal genomes, but without known homologs in other organisms. These 17 core, fungal genes were not found to differ from other yeast genes in GC content or codon usage patterns. More intensive study is required of these 17 genes and other common fungal genes to discover unique features of fungi compared to other organisms. PMID- 15883883 TI - Universal sharing patterns in proteomes and evolution of protein fold architecture and life. AB - Protein evolution is imprinted in both the sequence and the structure of evolutionary building blocks known as protein domains. These domains share a common ancestry and can be unified into a comparatively small set of folding architectures, the protein folds. We have traced the distribution of protein folds between and within proteomes belonging to Eukarya, Archaea, and Bacteria along the branches of a universal phylogeny of protein architecture. This tree was reconstructed from global fold-usage statistics derived from a structural census of proteomes. We found that folds shared by the three organismal domains were placed almost exclusively at the base of the rooted tree and that there were marked heterogeneities in fold distribution and clear evolutionary patterns related to protein architecture and organismal diversification. These include a relative timing for the emergence of prokaryotes, congruent episodes of architectural loss and diversification in Archaea and Bacteria, and a late and quite massive rise of architectural novelties in Eukarya perhaps linked to multicellularity. PMID- 15883884 TI - Tertiary windowing to detect positive diversifying selection. AB - As a protein-encoding gene evolves, different selective pressures act on the gene temporally and spatially. An examination of the ratio of nonsynonymous-to synonymous nucleotide substitution rate ratios (K(a)/K(s)) has proven to be a valuable method to examine selective pressures on protein encoding genes, including detecting positive diversifying selection. To gain power over averaging all sites in a gene together, examination of sites in primary sequence windows has frequently been employed. However, selection acts on folded proteins and sites that are close in tertiary space may not be close in primary sequence. A new method for the examination of K(a)/K(s) ratios based upon windows in tertiary structure is introduced and applied to the leptin gene family in mammals. Tertiary sequence windowing detects new sites under positive diversifying selection and detects positive diversifying selection with a more significant signal along various branches of the leptin gene family tree. PMID- 15883885 TI - Intron analyses reveal multiple calmodulin copies in Littorina. AB - Intron 3 and the flanking exons of the calmodulin gene have been amplified, cloned, and sequenced from 18 members of the gastropod genus Littorina. From the 48 sequences, at least five different gene copies have been identified and their functionality characterized using a strategy based upon the potential protein product predicted from flanking exon data. The functionality analyses suggest that four of the genes code for functional copies of calmodulin. All five copies have been identified across a wide range of littorinid species although not ubiquitously. Using this novel approach based on intron sequences, we have identified an unprecedented number of potential calmodulin copies in Littorina, exceeding that reported for any other invertebrate. This suggests a higher number of, and more ancient, gene duplications than previously detected in a single genus. PMID- 15883886 TI - Long inversely oriented subunits form a complex monomer of Tribolium brevicornis satellite DNA. AB - Highly abundant satellite DNA named TBREV is detected and characterized in the beetle Tribolium brevicornis (Insecta: Coleoptera). An outstanding peculiarity of the TBREV satellite monomer is its complex structure based on the two approximately 470-bp-long subunits, inversely oriented within a 1061-bp-long monomer sequence. The proposed evolutionary history demonstrates a clear trend toward increased complexity and length of the TBREV satellite monomer. This tendency has been observed on three levels: first as direct and inverted duplications of short sequence motifs, then by inverse duplication of the approximately 470-bp sequence segment, and, finally, by spread of inversely duplicated elements in a higher-order register and formation of extant monomers. Inversely oriented subunits share a similarity of 82% and have a high capacity to form a thermodynamically stable dyad structure that is, to our knowledge, the longest ever described in any satellite monomer. Analysis of divergences between inversely oriented subunits shows a tendency to a further reduction in similarity between them. Except in its centromeric localization, the TBREV satellite does not show similarity to other known Tribolium satellites, either in nucleotide sequence or in monomer length and complexity. However, TBREV shares common features of other Tribolium satellites that might be under functional constraints: nonconstant rate of evolution along the monomer sequence, short inverted repeats in the vicinity of an A+T tract, nonrandom distribution of A or T >/=3 tracts, and CENP-B box-like motifs. Although long inverted subunits might reinforce structural characteristics of the satellite monomer, their nucleotide sequence does not seem to be under constraints in order to preserve the dyad structure. PMID- 15883887 TI - Evolution of the mitochondrial genome in cephalochordata as inferred from complete nucleotide sequences from two epigonichthys species. AB - Complete mitochondrial (mt) DNA sequences of two lancelets, Epigonichthys maldivensis and E. lucayanus, were compared with those of two Branchiostoma lancelets and several deuterostomes previously surveyed. The mt-gene order of E. lucayanus was quite different from that of E. maldivensis, the latter being identical to the two Branchiostoma species. A remarkable genomic change in E. lucayanus mtDNA was an inversion, indicating the possibility of recombination of the mt-genome. Gene rearrangements, probably attributable to tandem genome duplications and subsequent random deletions, were observed in two parts. Short major unassignable sequences of the examined lancelets were regarded as a part of putative regulative elements, judging from some sequence similarity to the conserved sequence block (CSB) in mammalian mtDNA. The considerable mt-genome reorganization in E. lucayanus seemed to have affected the nucleotide substitution pattern, suggested by base composition analyses. The present analysis also suggested that AGR codons in lancelet mtDNA were likely to correspond to serine residue, rather than glycine. Furthermore, the AGG codon, so far reputed to be unassignable in lancelet mtDNA, was found twice in E. maldivensis, indicating the availability of all four AGN codons in some lancelets. This finding lends support to an alternative hypothesis regarding the evolutionary history of AGR-codon assignment in extant chordates, rather than that previously proposed. A molecular phylogenetic tree of the Epigonichthys and Branchiostoma species based on DNA sequences of the 13 mt-protein genes doubted the monophyly of the former genus, unlike the prevailing classification based on their different gonadal arrangements. PMID- 15883888 TI - Phylogeny of prokaryotes and chloroplasts revealed by a simple composition approach on all protein sequences from complete genomes without sequence alignment. AB - The complete genomes of living organisms have provided much information on their phylogenetic relationships. Similarly, the complete genomes of chloroplasts have helped to resolve the evolution of this organelle in photosynthetic eukaryotes. In this paper we propose an alternative method of phylogenetic analysis using compositional statistics for all protein sequences from complete genomes. This new method is conceptually simpler than and computationally as fast as the one proposed by Qi et al. (2004b) and Chu et al. (2004). The same data sets used in Qi et al. (2004b) and Chu et al. (2004) are analyzed using the new method. Our distance-based phylogenic tree of the 109 prokaryotes and eukaryotes agrees with the biologists "tree of life" based on 16S rRNA comparison in a predominant majority of basic branching and most lower taxa. Our phylogenetic analysis also shows that the chloroplast genomes are separated to two major clades corresponding to chlorophytes s.l. and rhodophytes s.l. The interrelationships among the chloroplasts are largely in agreement with the current understanding on chloroplast evolution. PMID- 15883889 TI - Phage associated bacteriocins reveal a novel mechanism for bacteriocin diversification in Klebsiella. AB - Ninety-six isolates of Klebsiella pneumoniae and K. oxytoca were recovered from wild mammals in Australia. 14.6% of these bacteria produce killing phenotypes that suggest the production of bacteriocin toxins. Cloning and sequencing of the gene clusters encoding two of these killing phenotypes revealed two instances of a bacteriocin associated with a bacteriophage gene, the first such genetic organization described. The newly identified klebicin C gene cluster was discovered in both K. pneumoniae and K. oxytoca. The newly identified klebicin D gene cluster was detected in K. oxytoca. Protein sequence comparisons and phylogenetic inference suggest that klebicin C is most closely related to the rRNase group of colicins (such as colicin E4), while klebicin D is most closely related to the tRNase group of colicins (such as colicin D). The klebicin C and D gene clusters have similar genetic and regulatory organizations. In both cases, an operon structure is inferred consisting of a phage-associated open reading frame and klebicin activity and associated immunity genes. This novel bacteriophage/bacteriocin organization may provide a novel mechanism for the generation of bacteriocin diversity in Klebsiella. PMID- 15883891 TI - More or less? PMID- 15883890 TI - Death pronouncements. PMID- 15883892 TI - Health care information provided by internet search engines. PMID- 15883893 TI - The CDC model of clinical instruction. PMID- 15883894 TI - Teaching medical students research while reaching the underserved. AB - OBJECTIVES: This study's objective was to develop and evaluate the effectiveness of a 9-week summer training program for teaching medical students research, while performing community-based participatory research in an underserved area. DESCRIPTION: Interactive didactic sessions familiarized students with research methods. Concurrently, they designed and completed a participatory project with a community mentor. EVALUATION: Questionnaires were used to assess students' experience with the program, their project, and their mentors. Pretests and posttests assessed students self-perceived understanding of research principles. CONCLUSIONS: Evaluation indicated that the program increased understanding of the research process and acquainted students with caring for patients in medically underserved communities. PMID- 15883895 TI - "I'm afraid of cancer". PMID- 15883896 TI - Bearing witness and healing through creativity. PMID- 15883897 TI - Research in residency: do research curricula impact post-residency practice? AB - BACKGROUND: The Residency Review Committee for Family Medicine recommends that research during residency be encouraged as a means of preparing residents for lifetime self-education after residency. No studies, however, have demonstrated the influence of these curricula on postgraduate practice. This study identified differences in attitudes, knowledge, or behavior between residency graduates who were or were not exposed to research training during residency. METHODS: A survey was mailed to 234 graduates of 13 family medicine residency programs in Michigan; eight of the programs included research curricula. The survey included questions on research training, confidence in reading medical articles, sources used for answering clinical questions, and current teaching and research activities. Responses were compared between graduates from programs with and without formal research training. RESULTS: The response rate was 53%. Graduates with research training reported significantly greater appreciation for research and its importance in guiding treatment decisions. There were no differences between groups in knowledge of statistics, comfort with reading research articles, or sources used for answering clinical questions, with the exception of greater use of Medline searches for research-trained residents. There were also no differences between groups in additional training or research experiences. Graduates with no formal research training reported being more involved after residency with student teaching and practice audits. CONCLUSIONS: Formal research training during residency appears to alter attitudes toward research and searches of the medical literature but does not to lead to greater participation in teaching or practice audits. PMID- 15883898 TI - Association between Balint training and physician empathy and work satisfaction. AB - BACKGROUND AND OBJECTIVES: Balint training is used in many family medicine residency programs to encourage self-reflection and exploration of the physician patient encounter. There is limited objective research, however, on outcomes of Balint training. This study's purpose was to determine whether Balint training is associated with differences in physician empathy and work satisfaction. METHODS: The Jefferson Scale of Physician Empathy and a validated survey on physician work satisfaction were mailed to 182 graduates of the Medical University of South Carolina (MUSC) Family Medicine Residency Program (113 Balint attendees and 69 nonattendees). The Dillman method of survey design was followed. RESULTS: The two groups were not statistically different in demographic measures. Balint attendees had a similar mean empathy score (119.4) as nonattendees (116.7). There was also no statistically significant difference in overall work satisfaction or satisfaction with financial compensation. Balint attendees were, however, more likely to say they would choose the same specialty if they could choose again than did nonattendees (86.1% versus 55%). CONCLUSIONS: There was no association found between Balint training and physician empathy, financial satisfaction, or overall work satisfaction. However, physicians who completed Balint training at the MUSC Family Medicine Residency Program seemed more satisfied with their choice of family medicine as a specialty. PMID- 15883899 TI - Factors associated with higher clinical skills experience of medical students on a family medicine preceptorship. AB - BACKGROUND AND OBJECTIVES: Educators should document students' clinical skills experience and whether curriculum and other course changes impact this experience. We wanted to determine the factors associated with students' clinical skills experience during their third-year family medicine preceptorship. METHODS: We conducted an observational study of 1,419 third-year students at the University of Iowa taking a required family medicine preceptorship during 9 academic years: 1994-1995 to 2002-2003. Students rated their level of experience with 57 clinical skills on a 5-point scale. Student ratings were summed to obtain scores for total clinical skills (n=57), procedural skills (n=22), and female specific skills (n=9). RESULTS: Students who completed their preceptorship in rural areas (as opposed to urban areas or in a residency), prior to the implementation of Centers for Medicare and Medicaid Services regulations, prior to integration with ambulatory internal medicine and community-based primary care, or who rotated during the second half of the academic year reported a higher mean level of experience with clinical skills in univariate and multivariate analyses. Both years of experience as a preceptor and total number of previous students taught were positively associated with students' total clinical skills scores. In addition, several preceptor teaching qualities were significantly and positively associated with overall skills experience: opportunity for clinical procedures, delegation of appropriate responsibility, and preceptor conveyed expectations clearly. CONCLUSIONS: Use of a consistent clinical skills checklist allowed us to document important effects of curriculum, legislation, and preceptor teaching qualities on the students' clinical skills experience. PMID- 15883900 TI - Cancer and colorectal cancer: knowledge, beliefs, and screening preferences of a diverse patient population. AB - BACKGROUND: Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States, with African Americans having the highest incidence and mortality of all racial and ethnic groups. CRC screening is widely recommended but remains underused, especially in minority populations. This study's purpose was to enhance our understanding of factors contributing to low screening rates among patients from a variety of racial and ethnic groups. METHODS: We conducted individual interviews with 30 participants, ages 50 or above, with an equal number of African Americans, Hispanics, and whites at a university-based family medicine clinic. We used open-ended interviewing techniques to elicit patient knowledge and beliefs regarding cancer, CRC, screening, and CRC screening tests. RESULTS: All groups, but particularly minority groups, lack knowledge of cancer, CRC, and screening. They did not understand the concept of screening, had difficulty listing common cancer and CRC screening tests, and had trouble understanding simplified medical terms and procedure names. Patients were hopeful about the benefit of early cancer diagnosis but remained reluctant to get tested if they are symptom free. CONCLUSIONS: Lack of understanding of cancer, screening, and routine terminology is a barrier to CRC screening, especially among minority groups. Effective communication strategies that address these issues may help increase CRC screening rates. PMID- 15883901 TI - The influence of academic projects on the professional socialization of family medicine faculty. AB - BACKGROUND: Successful academic physicians acquire and maintain productive colleague relationships, understand unwritten rules of academe, and effectively manage their careers. Developing these professional socialization skills are goals of some faculty development programs (FDPs), but there is scant evidence about whether such programs are effective. METHODS: A nationwide retrospective, cross-sectional written survey was conducted in two phases: (1) FDP directors who received US Department of Health and Human Services, Health Resources and Services Administration support between 1994 and 1997 described program activities and provided enrollee rosters and (2) enrollees reported socialization and colleague outcomes. Instruments were developed, pilot tested, and administered for this study. Analysis utilized descriptive statistics, factor analysis, and ANOVA. RESULTS: Of 52 eligible directors, 37 (71%) provided FDP activity details and rosters. Of 543 eligible enrollees, 351 (65%) returned surveys. A key result of factor analysis was a seven-item scale related to academic project activities. FDPs with greater emphasis on these activities were associated with enrollees reporting higher levels of colleague relationships and professional socialization skills. CONCLUSION: This study's factor analysis indicates that certain FDP project activities are positively associated with enrollees' professional socialization outcomes. PMID- 15883902 TI - The challenge of clinical interviewing and physical examination performance for general practitioners in Turkey. AB - BACKGROUND AND OBJECTIVES: This study's objective was to better understand the current conditions and style of practice of generalist physicians in Turkey on clinical interviewing and physical examination skills prior to the widespread availability of family medicine postgraduate training. METHODS: This study was performed in 30 primary health centers, randomly chosen from the 110 primary health centers in Izmir, Turkey. We administered a questionnaire to 106 physicians in those centers, asking about their performance during medical encounters. We then observed 166 first-visit encounters of 37 of those physicians. Finally, we compared physicians' reported behavior (as described in the questionnaires) with their actual performance (when observed). RESULTS: In the physician questionnaire, 86.8% of participants agreed that 20 minutes or more was sufficient time for first visits, but in practice, 81.9% of the interviews lasted less than 5 minutes. The major reason cited by physicians for short interview times was overcrowding (72.6%). In 94.6% of the encounters, physicians obtained the history of present illness but the rest of the history, such as past history and family history, was gathered in less than 40% of the interviews. Except for the examination of oral cavity and pharynx, lungs, and heart, almost all the rest of the physical examination was performed in less than 10% of the cases. No written records were kept in 63.0 % of the encounters. CONCLUSIONS: The problems and difficulties present in delivering primary care in Turkey include the physicians' behavior in addition to working conditions. PMID- 15883903 TI - Understanding interobserver agreement: the kappa statistic. AB - Items such as physical exam findings, radiographic interpretations, or other diagnostic tests often rely on some degree of subjective interpretation by observers. Studies that measure the agreement between two or more observers should include a statistic that takes into account the fact that observers will sometimes agree or disagree simply by chance. The kappa statistic (or kappa coefficient) is the most commonly used statistic for this purpose. A kappa of 1 indicates perfect agreement, whereas a kappa of 0 indicates agreement equivalent to chance. A limitation of kappa is that it is affected by the prevalence of the finding under observation. Methods to overcome this limitation have been described. PMID- 15883904 TI - Why pregnancy care should be an essential part of family medicine training. AB - Some family medicine educators are arguing to eliminate pregnancy care as a required component of family medicine training since the majority of family physicians no longer perform deliveries, and many programs are having increasing difficulties in meeting this training requirement. The primary benefit of pregnancy care training is not to produce family physicians who all perform deliveries but to produce family physicians who are competent to provide comprehensive primary care to women and girls, including routine and preventive reproductive care. The training in pregnancy care helps to differentiate family medicine residencies from other primary care training programs by facilitating competency in a wide range of reproductive health care for nonpregnant women and for the primary nonreproductive health care of pregnant and postpartum women. Residencies offering pregnancy care services also enhance their ability to train residents in child care. Family medicine should continue to strive to improve this aspect of residency training instead of abandoning it. PMID- 15883905 TI - Point-Counterpoint: the argument for a 2-year versus a 4-year family medicine residency. PMID- 15883908 TI - DNA vaccines forum: how to enhance DNA vaccines safety and potency? PMID- 15883909 TI - Replicating oncolytic virus therapeutics - Third International Meeting. PMID- 15883910 TI - Antibody-based cancer drugs: developing antibody therapeutic approaches to deliver individualized cancer drugs. PMID- 15883911 TI - Lung cancer - Second Annual Winter Conference. PMID- 15883912 TI - Drug Discovery Technology Europe 2005 - IBC's Ninth Annual Conference and Exhibition. PMID- 15883913 TI - Advances in Diabetes Treatment 2005 - Second Annual Conference. PMID- 15883914 TI - Stem cell research - Third Annual Asia-Pacific Congress: clinical trials and commercial applications. PMID- 15883915 TI - Tissue repair and ulcer/wound healing - Institut Pasteur Euroconference: molecular mechanisms, therapeutic targets and future directions. PMID- 15883916 TI - Anglo-Swedish Medicinal Chemistry - second symposium. PMID- 15883917 TI - American Chemical Society - 229th National Meeting: carbohydrate vaccines. PMID- 15883918 TI - American Chemical Society - 229th National Meeting: bench to pilot plant. PMID- 15883919 TI - American Chemical Society - 229th National Meeting: general oral and poster sessions. PMID- 15883920 TI - Antibacterial Drug Discovery and Development Summit - Tenth Annual SRI Summit. PMID- 15883921 TI - Nanoparticle technology for drug delivery: from nanoparticles to cutting-edge delivery strategies - part I. PMID- 15883922 TI - Nanoparticle technology for drug delivery: from nanoparticles to cutting-edge delivery strategies - part II. PMID- 15883923 TI - Recombinant polyclonal antibodies: therapeutic antibody technologies come full circle. AB - Current antibody therapeutics can be grouped into two generations, each distinguished by a unique feature of the immune system: diversity and specificity. Antibodies from human blood (immunoglobulin) represent the first generation, and are characterized by the natural diversity of human antibody responses. The second generation consists of recombinant monoclonal antibodies (mAbs), which are characterized by high specificity toward a single, often well described antigen. The natural immune response comprises a plurality of specificities, many of which do not compete for binding, whereas molecules in a mAb all compete for binding to the same epitope. Thus, the epitope is more likely to become a limiting factor for mAb binding to complex targets compared with a polyclonal antibody. Also, epitope-escape by mutation or natural variation is less likely to be a problem for polyclonal antibodies. Technologies attempting to develop truly human recombinant antigen-specific polyclonal antibodies, such as the Sympress technology, are closing a natural circle between the first generations of antibody technologies. PMID- 15883924 TI - Insomnia. AB - The launch of eszopiclone by Sepracor Inc in April 2005 in the US has marked the long-anticipated advent of a new generation of hypnotics that promise safer treatment for chronic insomnia. Over the coming years more compounds are expected to be launched, offering improved daytime well being and fewer side effects compared with their predecessors that were launched in the early 1990s. This article examines the factors that will drive the uptake of the new insomnia drugs by the public, describes potential barriers to adoption of these drugs and provides ideas for future treatment approaches that may offer better targeting and management of possible underlying conditions rather than merely controlling a symptom. PMID- 15883925 TI - Lubiprostone. AB - Sucampo Pharmaceuticals Inc and Takeda Pharmaceutical Co Ltd are developing, a ClC-2 chloride channel activator and a bicyclic member of a series of fatty acid compounds, for the potential treatment of a number of gastrointestinal conditions, including constipation, irritable bowel syndrome, postoperative bowel dysfunction and opioid bowel dysfunction. PMID- 15883926 TI - Homozygous nonsense mutations in KIAA1279 are associated with malformations of the central and enteric nervous systems. AB - We identified, by homozygosity mapping, a novel locus on 10q21.3-q22.1 for Goldberg-Shprintzen syndrome (GOSHS) in a consanguineous Moroccan family. Phenotypic features of GOSHS in this inbred family included microcephaly and mental retardation, which are both central nervous system defects, as well as Hirschsprung disease, an enteric nervous system defect. Furthermore, since bilateral generalized polymicogyria was diagnosed in all patients in this family, this feature might also be considered a key feature of the syndrome. We demonstrate that homozygous nonsense mutations in KIAA1279 at 10q22.1, encoding a protein with two tetratrico peptide repeats, underlie this syndromic form of Hirschsprung disease and generalized polymicrogyria, establishing the importance of KIAA1279 in both enteric and central nervous system development. PMID- 15883929 TI - Osseointegration in irradiated cancer patients: an analysis with respect to implant failures. AB - PURPOSE: This study retrospectively evaluated implant survival of 631 osseointegrated implants installed in irradiated cancer patients over a 25-year period. PATIENTS AND METHODS: The files of 107 patients followed since 1979 were evaluated. Factors influencing implant survival as oncologic treatment, radiotherapy protocols, patient and implant related elements were analyzed. RESULTS: Compared with a control group of non-irradiated patients, implant failures were higher after previous radiotherapy. High implant failures were seen after high dose radiotherapy and a long time after irradiation. All craniofacial regions were affected, but the highest implant failures were seen in frontal bone, zygoma, mandible, and nasal maxilla. Lowest implant failures were seen in oral maxilla. The use of long fixtures, fixed retention, and adjuvant hyperbaric oxygen therapy decreased implant failures. Noncontributing factors to implant survival were gender, age, smoking habits, tumor type and size, surgical oncologic treatment, and osseointegration (OI) surgery experience. CONCLUSION: Survival after cancer therapy is so high, and outcome from OI therapy so favorable that OI in the irradiated patient can be recommended. However, the OI clinician should be aware of the risks and pitfalls of treating such patients. PMID- 15883930 TI - Anchoring dental implant in tissue-engineered bone using composite scaffold: a preliminary study in nude mouse model. AB - PURPOSE: The purpose of this study was to fabricate a tissue-engineered bone graft anchoring dental implant with bone marrow stromal cell (bMSC) seeded coral implant composite scaffold. MATERIALS AND METHODS: Titanium dental implants (3 mm in diameter) were inserted into the cylinder coral scaffolds (5 mm in diameter and 1 mm in wall thickness). bMSCs were isolated from iliac bone marrow of adult New Zealand White rabbits, induced by dexamethasone and seeded into the composite scaffold at the density of 2 x 10 8 /mL in 200 muL medium. Nine cell coral implant complexes were incubated in vitro for 5 days. One complex was processed for scanning electronic microscopy. The other 8 complexes, together with 4 coral scaffold without cell acting as control, were implanted subcutaneously into nude mice back. At 1 and 2 months after implantation, 4 specimens from the experiment group and 2 specimens from the control group were harvested respectively. New bone restoration and new bone integration with dental implant were evaluated by gross inspection, manual handling test, radiographic examination, and histologic observation. RESULTS: Specimens harvested at 2 months after implantation were red and similar to native bone. Manual handling test showed that dental implants were fixed in the newly formed bone. Radiographic examination showed that most of the coral scaffold had been absorbed. Bone density x-ray shadow could be observed around the dental implant. Histologic examination showed that large amount of new bone formed around the dental implants and integrated well with the implants in some area. In the control group no bone formation was observed both macroscopically and microscopically. CONCLUSION: The results of the study suggested that the tissue-engineered bone of bMSCs seeded natural coral-implant composite scaffold is promising for dental implant anchoring, which has positive implication for clinical jaw reconstruction. PMID- 15883931 TI - Rehabilitation of a hemophiliac with implants: a medical perspective and case report. AB - A patient suffering from classical hemophilia had previous surgery for ankylosis of the right temporomandibular joint. This was replaced by a costochondral graft and an overlay of temporalis muscle. A bilateral sagittal split was performed for a micrognathic mandible and a sleep apnea problem. That procedure solved the sleep apnea; however, it resulted in a prognathic mandible and an anterior open bite. The lower anterior teeth were periodontally involved with impaired alveolar support. The restricted opening of the oral cavity of 18 mm between maxillary and mandibular centrals and the potential danger of bleeding complicated the surgical and restorative procedures. The patient was prepared medically on each of 4 occasions with factor VIII replacement concentrate, and oral antifibrinolytic therapy (tranexamic acid). The treatment of choice was the extraction of the remaining lower incisors and their replacement with an implant-supported temporarily cemented retrievable fixed prosthesis. Serial extractions and chairside temporization provided the surgeon with precise guides for implant placement, and enabled the patient to enjoy unimpaired function through periods of healing and osseointegration. PMID- 15883932 TI - Internal distraction osteogenesis with a unidirectional device for reconstruction of mandibular segmental defects. AB - PURPOSE: To present the authors' clinical experience with unidirectional internal distraction devices to reconstruct mandibular defects. PATIENTS AND METHODS: Five patients have been treated with mandibular distraction osteogenesis to reconstruct different acquired segmental defects. These mandibular defects (35 to 80 mm in length) were reconstructed by means of internal distraction devices with a transcutaneous activator. All the patients underwent complete resection of the affected bone and immediate placement of the distraction device on the remaining mandible. RESULTS: The results' analysis was based on clinical observation, postoperative radiographs, histopathologic findings and 3-dimensional computed tomographic scans. Successful distraction osteogenesis was achieved in 3 cases. In 1 case, extensive intraoral exposure of the device was observed, resulting in a failure of the procedure. One patient died of distant metastases 4 months after the resective surgery. CONCLUSION: Because of the limited number of cases, this study is preliminary. However, considering the good experimental and clinical results, this new technique offers an alternative for patients with segmental mandibular defects in which, because of local or general reasons, a more aggressive procedure should be avoided. PMID- 15883933 TI - Prediction accuracy of computer-assisted surgical visual treatment objectives as compared with conventional visual treatment objectives. AB - PURPOSE: This present study used the conventional visualized treatment objectives (VTOs) as a tool to evaluate the predictive value of the Dolphin computer assisted VTOs. MATERIALS AND METHODS: Presurgical cephalometric tracing predictions generated by oral and maxillofacial surgeons and the Dolphin Imaging software were compared with the postsurgical outcome as seen on lateral cephalometric tracings. Sixteen measurements of the predicted and actual postsurgical hard tissue landmarks were compared statistically. RESULTS: A paired Student's t test showed that 7 measurements had statistically significant differences for the conventional VTOs (facial angle, P < .0001; AOC, P < .0001; SNB, P = .003; ANB, P = .004; U1-NA-degrees, P = .01; U1-NA-mm, P = .02; and N perp Pog, P < .0001), while 9 measurements were statistically significant ( P = <.0001) for Dolphin (facial angle, P = .0001; AOC, P = .005; SNB, P = .001; ANB, I = .04; U1-NA-degrees, P = .003; PogNB, P = .04; U1-NA-mm, P = .002; N perp Pog, P = .0001; UFH, P = .03; and LFH, P = .03). CONCLUSION: From these data, it appears that both VTOs demonstrated good predictive comparative outcome and are equally precise. PMID- 15883934 TI - Evaluation of taste sensation following tongue reconstruction by microvascular forearm free flap. AB - PURPOSE: The objective of this study was to examine the restoration process of taste perception in patients after reconstruction of the tongue using microvascular forearm flaps. PATIENTS AND METHODS: Twelve patients, aged 28 to 70 years with a mean of 59.6 years consisting of 9 men and 3 women, were enrolled in this study. Examination results based on the filter paper disk and whole mouth methods were evaluated on a 6-point rating scale. RESULTS: In none of the patients was it possible to detect any perception of the test tastes in the regions under the control of the chorda tympani and glossopharyngeal nerves on the reconstructed side, indicating a "nondetectable" grade 6 result. The evaluation results for the normal tongue region under the control of the chorda tympani in the patients after tongue reconstruction showed a mean grade of 4.7 for the sweet, 4.9 for the salty, 4.6 for the sour, and 4.4 for the bitter taste. Mean grade determined by the whole mouth method was 3.6 for sweet, 3.3 for salty, 3.0 for sour, and 3.6 for bitter. CONCLUSION: No taste perception was detected in the regions of the tongue reconstructed with forearm flaps. These results led us to assume that compensatory taste perception must occur in other regions of the oral cavity because none of our postsurgery patients showed any serious taste disorder. PMID- 15883935 TI - Hemoglobin value reduction and necessity of transfusion in bimaxillary orthognathic surgery. AB - PURPOSE: It has been the aim of the present clinical study to assess the reduction of the hemoglobin value and the frequency of blood transfusions during bimaxillary orthognathic surgery and to discuss the clinical consequences. PATIENTS AND METHODS: Fifty-six patients (31 female, 25 male; mean age, 28.6 +/- 13.0 years; range, 14 to 66 years) were operated on. Twenty-nine patients predeposited blood before surgery. As a threshold for intraoperative or postoperative transfusion, a hemoglobin value of 7.5 g/100 mL was chosen. RESULTS: Perioperatively, the hemoglobin values of the patients who predeposited blood decreased significantly after blood donation. The hemoglobin value reduced postoperatively by 2.6 +/- 1.4 g/100 mL in the non-donors and by 2.6 +/- 1.1 g/mL in the donors. None of the patients who did not predeposit blood received homologous blood transfusions intraoperatively or postoperatively. In the group of patients who predeposited blood, 3 were transfused intraoperatively. They received 1 or 2 units of autologous blood. CONCLUSION: The individual statistics of the department show that there was only a limited reduction of the intraoperative and postoperative hemoglobin values as a consequence of bimaxillary orthognathic surgery. The increased safety of homologous blood and the minimal transfusion rates support abandonment of routine predepositing of autologous blood and the acceptance of homologous blood in the rare case of transfusion in bimaxillary surgery. PMID- 15883936 TI - Stability of the Le Fort I osteotomy with anterior internal fixation alone: a case series. AB - PURPOSE: The purpose of this case series was to evaluate the late postsurgical stability of the Le Fort I osteotomy with anterior internal fixation alone and no posterior zygomaticomaxillary buttress internal fixation. PATIENTS AND METHODS: Sixty patients with maxillary vertical hyperplasia and mandibular retrognathia underwent a 1-piece Le Fort I osteotomy of the maxilla with superior repositioning and advancement or setback. A bilateral sagittal split ramus osteotomy for mandibular advancement was also performed in 22 patients. Stabilization of each maxillary osteotomy was achieved using transosseous stainless steel wires and/or 3-hole titanium miniplates in the piriform aperture region bilaterally, with no zygomaticomaxillary buttress internal fixation. (Twelve of the 60 identified patients were available for a late postoperative radiographic evaluation.) Lateral cephalometric radiographs were taken preoperatively (T1), early postoperatively (T2), and late postoperatively (T3) to analyze skeletal movement. RESULTS: These 12 patients (5 male, 7 female) had a mean age of 24.5 years at surgery. Mean time from surgery to T2 was 41.2 days; mean time from surgery to T3 was 14.8 months. One patient received anterior wire osteosynthesis fixation, while 11 patients received both anterior titanium miniplate internal skeletal fixation and anterior wire osteosynthesis fixation. Six patients underwent Le Fort I osteotomy with genioplasty, 1 patient underwent Le Fort I osteotomy with bilateral sagittal split ramus osteotomy, and 5 patients underwent Le Fort I osteotomy with bilateral sagittal split ramus osteotomy and genioplasty. These 12 patients all underwent maxillary superior repositioning with either advancement (11 patients) or setback (1 patient). Statistically significant surgical (T2-T1) changes were found in all variables measured. In late postsurgical measurements (T3-T2), all landmarks in the horizontal and vertical plane showed statistically significant skeletal stability. CONCLUSION: This case series suggests that anterior internal fixation alone in cases of 1 piece Le Fort I maxillary superior repositioning with advancement has good late postoperative skeletal stability. PMID- 15883937 TI - A retrospective review of treatment of the odontogenic keratocyst. AB - PURPOSE: The purpose of this study was to evaluate different surgical treatment methods for odontogenic keratocysts and the outcome of those treatments over a 25 year period. PATIENTS AND METHODS: A retrospective review was performed of 40 patient charts treated at the University of Iowa Hospitals and Clinics (Iowa City, IA) from 1977 to 2002 with the diagnosis of odontogenic keratocyst. Demographic data were collected along with lesion location, symptoms present at initial presentation, surgical treatment rendered, length of follow-up, and incidence of recurrence. RESULTS: Surgical treatments included enucleation, enucleation with Carnoy's solution, peripheral ostectomy, peripheral ostectomy with Carnoy's solution, and en bloc resection. Recurrence was found in 9 to 40 patients. Seven of 9 recurrences (78%) occurred in 5 years or less, with 2 (22%) occurring more than 5 years after initial treatment. Patients treated with enucleation had a recurrence rate of 54.5% (6 of 11 patients). One of 2 patients treated with enucleation and Carnoy's solution had a recurrence. Those treated with peripheral ostectomy had a recurrence rate of 18.2% (2 of 11). Peripheral ostectomy with Carnoy's solution had no recurrences (0/13). CONCLUSION: Treatment of an odontogenic keratocyst with peripheral ostectomy, with or without the use of Carnoy's solution, had a significantly lower rate of recurrence. Treatment with enucleation, with or without the use of Carnoy's solution was associated with a significantly higher recurrence rate. PMID- 15883938 TI - Radiographic evaluation of the status of third molars in the Asian-Indian students. AB - PURPOSE: The purpose of this study is to assess the prevalence and agenesis of third molars, their impaction status, and the time and sequence of their eruption in the students of Asian-Indian population. PATIENTS AND METHODS: One hundred students with ages ranging from 17.5 to 20.0 years were randomly selected. This age group was selected because this is the age at which third molars normally begin to erupt. Students who had undergone any tooth extraction or orthodontic treatment previously were not included in the study. RESULTS: The incidence of congenital missing third molars was 11.5%, with a higher incidence in females. Of the third molars present, 31% were in mesioangular position, 42% were in the vertical position, 27% were in the distoangular position, and none were in the horizontal position. Of the 354 teeth examined, 24% of teeth were found to be erupted and 76% were in various stages of eruption. The level of eruption of third molars was 27% at A; 39%, B; and 34%, C. CONCLUSIONS: The present study showed that at the baseline (mean age, 19.3 years), 24% of third molars were erupted, 76% were in various stages of eruption, and 11.5% were congenitally missing in the Asian-Indian student population. PMID- 15883939 TI - Reconstruction of orbital floor fracture with polyglactin 910/polydioxanon patch (ethisorb): a retrospective study. AB - PURPOSE: We sought to evaluate the effectiveness and the complications related to the use of Ethisorb (resorbable alloplastic material) in the reconstruction of orbital floor fractures. PATIENTS AND METHODS: We retrospectively reviewed the charts of all patients who underwent orbital floor fracture reconstruction with Ethisorb since 2001. We only included patients with a minimum follow-up of 3 months. The following data were recorded for every patient: age, gender, cause of trauma, time from trauma to surgery, signs and symptoms, concomitant ocular injuries, radiographic analysis, pertinent intraoperative findings (including the type of approach), follow-up time, and postoperative complications. RESULTS: Eighty-seven patients were included in the study. Twenty-one patients (24.1%) experienced postoperative complications. Of these, only 3 patients (3.4%) had permanent complications directly related to the Ethisorb membrane (diplopia, enophthalmos). Two of these patients required revision surgery and are discussed in the article. CONCLUSIONS: The results of our study demonstrate the effectiveness of Ethisorb in the repair of small-to-moderate orbital floor fracture defects (up to a maximum size of 2 x 2 cm). PMID- 15883940 TI - Facial soft tissue injuries as an aid to ordering a combination head and facial computed tomography in trauma patients. AB - PURPOSE: While the trauma computed tomography (CT) head scan is frequently ordered during resuscitation of multiply injured patients, determining when also to order facial CT in these situations can be difficult. Consequently, facial fractures are commonly missed on initial imaging evaluation. In acute trauma situations, facial soft tissue injury markers are often used to aid in this decision. A study was undertaken to identify exactly which facial soft tissue injuries correlate with facial fractures and which did not. The aim of our research was to compare the facial soft tissue injury patterns of patients receiving a combination head and facial CT who had facial fractures versus those who had no facial fractures. PATIENTS AND METHODS: A retrospective trauma registry review was performed for a 5-year period at Legacy Emanuel Hospital, a level 1 trauma center. The trauma team saw a total of 9,871 trauma patients and 49.9% required head CT as part of their initial evaluation. A total of 777 (15.7%) patients had a combination head and facial CT in which 477 (61.4%) had a facial fracture and 300 (38.6%) had no facial fracture. Statistical significance was established using Bonferroni corrected P values less than .05. RESULTS: Lacerations occurring in areas of the lips, nose, and intraorally, as well as wounds leading to periorbital contusion and subconjunctival hemorrhage, were significantly prevalent in the fracture group. Conversely, scalp lacerations and scalp contusions were significantly higher in the nonfracture group. Other injuries such as lacerations of the tongue, chin, forehead, cheek, ear, eyelid, and eyebrow were indistinguishable between both groups. CONCLUSIONS: Our data demonstrate that there are certain facial soft tissue injury zones that correlate with facial fractures in trauma patients who obtain head CT. We would like to propose that the acronym LIPS-N (Lip laceration, Intraoral laceration, Periorbital contusion, Subconjunctival hemorrhage, and Nasal laceration) be used in conjunction with a physical examination when assessing if a trauma patient who is getting head CT should also get facial CT. PMID- 15883941 TI - Treatment modalities for mandibular angle fractures. AB - PURPOSE: Management of mandibular angle fractures is often challenging and results in the highest complication rate among fractures of the mandible. Optimal treatment for angle fractures remains controversial. Historically, treatment of mandible fractures included intraoperative maxillomandibular fixation (MMF) along with rigid internal fixation. More recently, noncompression plates miniplates, which produce only relative stability, have gained popularity. The absolute necessity of intraoperative MMF as an adjunct to internal fixation has also become controversial. The current trends in the management of simple, noncomminuted mandibular angle fractures are examined. MATERIALS AND METHODS: A survey was submitted to North American and European AO ASIF (Arbeits-gemeinschaft fur Osteosynthesefragen Association for the Study of Internal Fixation) faculty in July 2001. Statistical analysis of results included both Fisher's exact and chi-square tests. Results were considered significant if P <.05. RESULTS: One hundred ten of 127 potential responses were received (87%). Among 104 surgeons who treat mandible fractures, 86 (83%) treat more than 10 mandibular fractures per year. Preferred techniques for simple, noncomminuted mandibular angle fractures in this group were: single miniplate on the superior border (Champy technique) with or without arch bars (44 surgeons, 51%); tension band plate on the superior border and nonlocking, bicortical screw plate on the inferior border (11 surgeons, 13%); dual miniplates (9 surgeons, 10%); a locking screw plate on the inferior border only (6 surgeons, 7%), and 3-dimensional plates (5 surgeons, 6%). Eleven surgeons (13%) gave multiple answers. Although only 13% of surgeons surveyed primarily use the combination of tension band and nonlocking, bicortical screw plates, many surgeons (73%) continue to use this technique in certain circumstances. Within this group, 32 (51%) place screws in a neutral position, while 31 (49%) place screws in an eccentric position, resulting in compression. For simple noncomminuted angle fractures, the number of surgeons performing internal fixation without MMF were: 14 often (16%); 20 occasionally (23%); 17 seldom (20%); and 35 never (41%). Surgeons treating more than 10 versus those who treat less than 10 fractures per year, International versus North American faculty, and Oral and Maxillofacial surgeons (OMS) versus non-OMS surgeons were compared. Surgeons who treat more than 10 fractures per year favor the Champy technique over the tension band and bicortical plate combination (44 [51%] vs 11 [13%]), while those surgeons who treat less than 10 per year favor the tension band and bicortical plate combination over the Champy technique (9 [50%] vs 3 [17%]; P < .01, Fisher exact test). International faculty are less likely to use intraoperative MMF than North American faculty (29 [81%] vs 31 [43%]; P < .01, Fisher exact test). OMS surgeons are less likely to use the tension band and bicortical plate combination than non-OMS surgeons (22 [56%] vs 42 [90%]; P < .017, Fisher exact test). CONCLUSION: This survey suggests an evolution in the management of mandibular angle fractures. A single miniplate plate on the superior border of the mandible has become the preferred method of treatment among AO faculty. When using large, inferiorly based plates more surgeons are now favoring neutral rather than eccentric screw placement. Intraoperative MMF is not considered mandatory by some surgeons in certain circumstances. PMID- 15883942 TI - Mandibular distraction osteogenesis: a rabbit model using a novel experimental design. AB - PURPOSE: Distraction osteogenesis (DO) is a surgical procedure that targets bone regeneration and elongation, currently used in the treatment of many craniofacial deformities. The quest for optimization of DO clinical parameters has led to the development of a variety of animal models. Our study aims to establish a rabbit animal model of mandibular DO, in which the control osteotomy and distraction device are placed on the opposite hemimandible from the one being distracted, within the same animal host. Furthermore, we propose to histologically characterize the different stages or distraction and consolidation in the same animal model. MATERIALS AND METHODS: Twenty-five rabbits underwent mandibular osteotomies and bilateral placement of distraction devices. After a latency of 3 days, the distraction device was activated on one side of each animal at a rate of 0.5 mm/12 hours for 7 days, while the other side remained inactive (control). This was followed by a consolidation period of 14 days. Five animals per time point were killed on days 3, 7, 10, 17, and 24. RESULTS: Gross tissue analysis showed a 7-mm callus formation at the distracted side and a well-healed osteotomy in the non-distracted side. Clinically, a unilateral Class III malocclusion occurred in the distracted side. Histology at each time-point shows new bone formation and orientation of the bony spicules along the axis of the mechanical strain. CONCLUSIONS: We have established and characterized an animal model of mandibular DO that outlines valid biologic controls and provides thorough monitoring of the DO process. PMID- 15883943 TI - Analysis of cell-seeded 3-dimensional bone constructs manufactured in vitro with hydroxyapatite granules obtained from red algae. AB - PURPOSE: Bone tissue engineering is a promising approach for the treatment of defective or lost bone in the maxillofacial region. Biocompatible and biodegradable scaffolds seeded with living cells are used to create functional tissue for load-bearing bone reconstruction. The aim of this study was to manufacture cell-seeded 3-dimensional bone constructs based on hydroxyapatite ceramic granule calcified from red algae and mesenchymal cambial-layer precursor cells. The ability of these cells to grow on hydroxyapatite ceramic was quantitatively investigated to evaluate 3-dimensional bone constructs for their potential use in bone tissue engineering. MATERIALS AND METHODS: Mesenchymal cambial-layer precursor cells were isolated from mandibular periosteum biopsy samples of 3 patients. To manufacture 72 bone constructs, these cells and hydroxyapatite ceramic granules (C GRAFT/Algipore; Clinician's Preference LLC, Golden, CO) were cultivated under osteogenic differentiation conditions in a rotating wall vessel system. After 6 and 21 days, histologic examination and scanning electron microscopy were performed. The absolute DNA content, protein synthesis, and alkaline phosphatase activity were also quantified. The osteoblastic phenotype of the constructs was confirmed by the expression of bone specific genes (osteocalcin, osteonectin, osteopontin, and core binding factor alpha1) using semiquantitative reverse transcription-polymerase chain reaction and Western blot analysis. RESULTS: Cells within the constructs showed good viability, which was evidenced by an increase in DNA content over the culture period. The decrease in alkaline phosphatase-specific activity could be an indicator of the maturation of cells and the induction of mineralization. The osteoblastic phenotype of the constructs was demonstrated on protein and at the RNA level over the entire culture period. CONCLUSION: We observed a positive effect of hydroxyapatite ceramic granules on mesenchymal cambial-layer precursor cell behavior in cell-seeded 3-dimensional bone constructs, indicating the potential applicability of C GRAFT/Algipore composites in bone tissue engineering. PMID- 15883944 TI - Bisphosphonate osteochemonecrosis (bis-phossy jaw): is this phossy jaw of the 21st century? AB - PURPOSE: Bisphosphonates are being implicated in a growing number of complications of the jaws. A number of terms are being applied to this phenomenon and perhaps the descriptive term bisphosphonate osteochemonecrosis has the most merit. But the eerie similarity of this 21st century disease process with the 19th century disease known as phossy jaw is striking. As the nomenclature continues to evolve, the term used in this article will be bis-phossy jaw. This article will explore historical and current aspects of these diseases. Although there may be other mitigating factors, such as oral health, chemotherapy history, immune status, Karnofsky performance status, or Kaplan-Feinstein index, bisphosphonates appear to be the necessary component in cases of bis-phossy jaw. MATERIALS: This is primarily a review article on reported cases of bis-phossy jaw, with historical looks at phossy jaw and osteoradionecrosis. Our laboratory has reviewed 20 suspected cases of bis-phossy jaw and the typical histopathologic features of bis-phossy jaw are presented. RESULTS: Descriptions of phossy jaw and current bis-phossy jaw cases are remarkably similar. Histopathologic features of bis-phossy jaw showed intact vascular channels, even in areas with acute inflammatory infiltrates and bacterial overgrowth. Non-vital bone fragments with reduced evidence of osteoclastic action were also noted. CONCLUSION: Bis-phossy jaw may have more of a bacterial cofactor risk than osteoradionecrosis, and though altered angiogenesis may yet prove to be a factor, avascularity does not appear to be a major cofactor. The historical disease phossy jaw appears to serve as a possible analogous disease for current research and treatment of bis-phossy jaw. Prevention and early identification of patients at risk should be of prime concern. PMID- 15883945 TI - Kimura's disease: report of a case with presentation in the cheek and upper eyelid. PMID- 15883946 TI - The glandular odontogenic cyst: clinicopathologic features and treatment of 14 cases. PMID- 15883947 TI - Periosteal osteosarcoma of the mandible. PMID- 15883948 TI - Melanotic schwannoma arising in the floor of the mouth. PMID- 15883949 TI - Horner's syndrome due to a large schwannoma of the cervical sympathetic chain: report of case. PMID- 15883950 TI - Surgically assisted rapid palatal expansion using a new distraction device: report of a case with an epimucosal fixation. PMID- 15883951 TI - Surgical approach to the stylohyoid process in Eagle's syndrome. PMID- 15883952 TI - Intraoperative video monitoring with a wireless micro digital camera system. PMID- 15883953 TI - Fixation of comminuted alveolar ridge fracture after severe trauma by application of ti-osteosynthesis plates on soft tissue: a technical note. PMID- 15883954 TI - Coronectomy: a questionable procedure. PMID- 15883955 TI - Bone distraction versus dynamic guided bone regeneration. PMID- 15883956 TI - Acute Dialysis Quality Initiative (ADQI): the PASSPORT project. PMID- 15883957 TI - The 3rd International Consensus Conference of the Acute Dialysis Quality Initiative (ADQI). PMID- 15883958 TI - Application of blood purification to non-renal organ failure. AB - The application of artificial organs to the task of blood purification in the setting of non-renal organ failure simultaneously presents important challenges and opportunities. Failures of cardiovascular, hepatic, coagulation and immune systems are all characterized by dysregulation leading to multi-organ failure. When sustained, these conditions result in multiple organ system dysfunction and death and are far too common in modern intensive care units (ICUs). While the pathogenesis of each of these organ failures is complex and variable, brought about by a variety of underlying conditions, the potential to improve patient outcomes by simultaneously targeting multiple pathways can perhaps best be realized by blood purification. Unlike drug strategies, which are usually limited to one component of these complex networks, blood purification is, by its very nature, broad spectrum and self regulating. For example, as the concentration of mediators or toxins increases, so does removal. Furthermore, given the many failed trials of specific therapy, the recent focus of immunomodulatory therapy in sepsis has shifted to non-specific methods of influencing the entire inflammatory response without suppressing it. In this issue of the journal, members of the Acute Dialysis Quality Initiative (ADQI) present systematic reviews on the application of hemofiltration, ultrafiltration, plasma therapies and liver-assist therapy for the treatment of non-renal organ failure. The focus of these reviews is on clinical evidence as well as recommendations for future research. PMID- 15883959 TI - Extracorporeal blood treatment (EBT) methods in SIRS/Sepsis. AB - BACKGROUND: Extracorporeal blood purification treatment (EBT) methods have been used in the treatment of experimental and human SIRS/Sepsis in a variety of settings and with variable reports of efficacy and safety. Their role in the management of SIRS/Sepsis remains controversial. OBJECTIVES: To develop consensus statements regarding important aspects of research, practice and technical management concerning EBT. METHODS: Systematic review of published study. Evidence-based grading of information available. Consensus development regarding fundamental questions about EBT. RESULTS: Consensus was achieved on all questions posed during the conference. It was agreed that there is currently a clear biological rational for EBT in SIRS and Sepsis. It was agreed that conventional CVVH has sufficiently been shown not to be effective in SIRS/Sepsis in the absence of concomitant ARF and that other therapies such as plasmapheresis or high-volume hemofiltration or coupled plasma filtration adsorption appear more promising and should be tested in multicentre randomized controlled trials. Patients with ARF and SIRS/Sepsis should be treated differently from those with ARF alone even though current practice in this field is not well known. Patients with refractory septic shock should be considered for EBT. Appropriate end points for clinical trials can be defined and chosen according to the goals of the trial. Different technologies exist for EBT and better understanding of the merits and safety of each is needed as well as better standardization of methodology and dose. CONCLUSIONS: Broad consensus can be achieved on several aspects of EBT and can be used to inform clinical practice and to help guide the establishment of a future research agenda. PMID- 15883960 TI - Plasma therapies in thrombotic syndromes. AB - BACKGROUND: Plasma therapies are being applied to thombotic syndromes, but there are limited controlled studies. OBJECTIVE: To review the evidence and the current practices for plasma therapies in thrombotic syndromes. METHODS: Expert-enhanced evidence-based analysis. Evidence obtained as of Dec 31, 2002 using PubMed electronic reference library and expert-obtained library for a total of > 3,000 references obtained using the terms plasma therapy or plasma exchange or plasmapheresis or plasmafiltration or sorbents each combined with the words thrombotic syndrome or sepsis or septic shock. The authors screened the abstracts, reviewed the agreed set of papers, and compiled the recommendations. RESULTS: Plasma therapies, which alter the plasma components in patients, have been applied in thrombotic syndromes worldwide. In these patients, there is a biologic plausibility for plasma therapies since they have molecules that are prothrombotic and/or antifibrinolytic which would put them at risk for microvascular thrombosis and end-organ damage. There are respectively one randomized controlled trial (RCT) in primary thrombotic syndrome, and secondary thrombotic syndrome, which showed an improvement in mortality in applying plasma therapies (plasma exchange by centrifugation). However, there are numerous non randomized and case series. Plasma exchange is accepted as the standard therapy for primary thrombotic syndrome as in thrombotic thrombocytopenic purpura (TTP). However, no consensus has been reached for plasma exchange in secondary thrombotic syndromes such as in sepsis, hemolytic uremic syndrome (HUS), thrombocytopenia associated multiple organ failure, TTP/HUS, s/p bone marrow or solid organ transplant, HELLP syndrome, immunologic disorders, drug exposure, or pancreatitis. CONCLUSIONS: As we understand more about the pathophysiology of thrombotic syndromes, specific plasma therapies can be applied for the specific need of a particular patient population. There are sufficient preliminary data to recommend a definitive RCT to evaluate the efficacy of the different types of plasma therapies in secondary thrombotic syndromes. PMID- 15883961 TI - Extracorporeal ultrafiltration for acute exacerbations of chronic heart failure: report from the Acute Dialysis Quality Initiative. AB - This report from a work group affiliated with the Acute Dialysis Quality Initiative is a critical assessment of the use of extracorporeal ultrafiltration (UF) in the management of acutely decompensated heart failure (HF). In addition to assessing UF in this setting, the report also provides background information on HF, including classification, pathophysiology, and the importance of concomitant renal failure. A summary of important results from clinical trials in this area is provided, along with a discussion of technical considerations. Finally, specific recommendations for future clinical evaluations are given. PMID- 15883962 TI - Liver support. PMID- 15883963 TI - Sequential convective therapies (SCT): a prospective study on feasibility, safety, adequacy and tolerance of on-line hemofiltration and hemodiafiltration in sequence. AB - Sequential dialysis techniques (i.e pure ultrafiltration followed by dialysis) have been used in the past, due to their capability to remove large volumes of fluids without inducing hemodynamic instability. The disadvantages of inadequate efficiency and lack of technology lead to the decline of such methods. Hemofiltration (HF) and hemodiafiltration (HDF) are recently being utilized in a greater proportion thanks to on-line fluid preparation systems. Each process (HF and HDF) has its own benefits in the removal of small, medium and high-molecular weight substances and in hemodynamic stability. Sequential convective therapies (SCT) such as hemofiltration-hemodiafiltration in sequence (HF-HDF) may combine the benefits and eliminate the disadvantages of each method and should be studied in order to explore their potential application in modern dialysis. Furthermore they can be easily applied nowadays, due to the development of new sophisticated dialysis machines. In order to evaluate the feasibility, safety, efficiency and tolerance of different SCT methods we studied 3 schedules: SCT1: 1h pre-dilution HF followed by 3h of post-dilution HDF (in the HF mode we lost 25% of the total fluid that had to be removed). SCT2: 1h pre-dilution HF followed by 3h of post dilution HDF (in the HF mode we lost 50% of the total fluid that had to be removed). SCT3: 2h pre-dilution HF followed by 2h of post-dilution HDF (in the HF mode we lost 50% of the total fluid that had to be removed). We studied 6 chronic hemodialysis patients using the same machine (AK200 ULTRA), with on-line fluid preparation system and the same type of dialyzer (Polyflux 210). SCT schedules were compared to on-line HF, on-line HDF and high flux dialysis performed with the same dialyzers. The treatments resulted safe, easy, feasible and well tolerated with an improved hemodynamic response to high volume convective therapies. Adequacy of treatment was satisfactory in all SCT schedules while middle molecular weight solute clearance and removal resulted higher in treatments with higher convective component. SCT might represent an interesting option for the future especially in patients with hemodynamic instability and requirements for interventions during treatment. PMID- 15883967 TI - The metabolic syndrome and high C-reactive protein: prevalence and differences by sex in a southern-European population-based cohort. AB - BACKGROUND: To evaluate the prevalence of the metabolic syndrome (MS) and its components in a population-based cohort, and to analyse the association between gender, environmental conditions, C-reactive protein (CRP), and the syndrome. METHODS: Out of 1877 subjects aged 45-64, who represented all the patients of six family physicians, representative of the sanitary districts of Asti (north western Italy), 88% accepted to participate in an interview on personal habits, and several clinical and laboratory measurements. RESULTS: The MS (National Cholesterol Education Program criteria) was present in 24% of males and 22% of females. Males had a significantly higher percentage of hyperglycaemia, hypertension, hypertriglyceridemia, whereas females had a higher prevalence of central obesity and low HDL-cholesterol. In a multiple logistic regression model, the MS was significantly associated with increasing age, BMI, and >30 g/day alcohol intake (OR = 1.42; 95% CI 1.27-1.58), and negatively to higher education level (OR = 0.52; 95% CI 0.28-0.99) and moderate exercise (OR = 0.65; 95% CI 0.57 0.76). CRP levels are highly correlated to BMI and the components of the syndrome. The association between CRP and the MS remains significant in women only, in a multivariate analysis, after multiple adjustments (OR = 1.73; 95% CI 1.42-2.11). Higher CRP levels, correlated to smoking and, inversely, to alcohol intake, identify a further 12% of the cohort at higher cardiovascular risk. CONCLUSIONS: The MS affects more than 20% of this middle-aged cohort, but more than 30%, with higher CRP levels are at high cardiovascular risk. Healthier lifestyle habits are inversely associated with the MS and CRP levels, suggesting the need for strategies and their implementation in the general population. PMID- 15883968 TI - Energy flow in push-pull chromophores: vibrational dynamics in para-nitroaniline. AB - para-Nitroaniline (PNA) plays an essential role as the prototype model of push pull chromophores. The nature and degree of participation of vibrational degrees of freedom in the charge-transfer and internal-conversion processes are current issues of great theoretical and practical importance. Ultrafast time-resolved anti-Stokes resonance Raman spectroscopy (TRARRS) experiments on PNA in dimethyl sulfoxide with three different excitation wavelengths were performed to probe these dynamical influences. The vibrational dynamics associated with S0 were independent of incident wavelength, and this supports the picture that the S1 dynamics are fast relative to the rate of intersystem crossing. The phenyl breathing mode nu(19) (860 cm(-1)) and the symmetric NO2 stretch nu(29) (1310 cm( 1)) exhibited vibrational lifetimes in S0 of 8.1 and 5.2 ps, respectively. No evidence for inhomogeneous broadening of the charge-transfer band in the UV/Vis absorption spectrum was found. PMID- 15883969 TI - Simultaneous determination of 1H-1H and 1H-13C residual dipolar couplings in a chiral liquid crystal solvent using a natural abundance HSQC experiment. AB - A high-resolution, phase-sensitive, natural abundance F2-coupled 1H-13C HSQC (F2HSQC) NMR experiment was developed to measure simultaneously both (n)D(HH) and 1D(CH) residual dipolar couplings (RDCs) of small molecules present in a chiral polypeptide liquid crystal solvent system composed of poly-gamma-benzyl-L glutamate (PBLG) in CDCl3. Because this is an indirect-detection NMR experiment, the relatively small amount of sample (7.5 mg in this study) and short acquisition times (5 h) that are required make this HSQC experiment well suited for samples that are either limited in solubility or in quantity or require short analysis times. The F2HSQC experiment can be performed without any specialized equipment or sample modification and can enhance our ability to measure RDCs accurately and rapidly in polypeptide liquid crystal solvents. PMID- 15883971 TI - Chemoenzymatic synthesis of HIV-1 gp41 glycopeptides: effects of glycosylation on the anti-HIV activity and alpha-helix bundle-forming ability of peptide C34. AB - C34 is a 34-mer peptide derived from the C-terminal ectodomain of HIV-1 envelope glycoprotein, gp41. The C34 region in native gp41 carries a conserved N-glycan at Asn637 and the sequence is directly involved in the virus-host membrane fusion, an essential step for HIV-1 infection. This paper describes the synthesis of glycoforms of C34 which carry a monosaccharide, a disaccharide, and a native oligosaccharide moiety. The synthesis of the glycopeptide which carries a native high-mannose type N-glycan was achieved by a chemoenzymatic approach by using an endoglycosidase-catalyzed oligosaccharide transfer as the key step. The effects of glycosylation on the inhibitory activity and the helix-bundle forming ability of C34 were investigated. It was found that glycosylation moderately decreases the anti-HIV activity of C34 and, in comparison with C34, glyco-C34 forms less compact six-helix bundles with the corresponding N-terminal peptide, N36. This study suggests that conserved glycosylation modulates the anti-HIV activity and conformations of the gp41 C-peptide, C34. PMID- 15883970 TI - Examples of peptide-peptoid hybrid serine protease inhibitors based on the trypsin inhibitor SFTI-1 with complete protease resistance at the P1-P1' reactive site. AB - Research in the field of protease inhibitors is focused on obtaining potent, specific and protease-resistant inhibitors. To our knowledge, there are no reports in the literature that consider the application of N-substituted glycine residues (peptoid monomers) for the design of peptidomimetic protease inhibitors. We hereby present the chemical synthesis and kinetic properties of two new analogues of the trypsin inhibitor SFTI-1 modified at the P1 position. Substitution of Lys5 in SFTI-1 by N-(4-aminobutyl)-glycine and N-benzylglycine, which mimic Lys and Phe, respectively, made these analogues completely protease resistant at their P1-P1' reactive sites. The analogues synthesised appeared to be potent inhibitors of bovine beta-trypsin and alpha-chymotrypsin. These noncovalent, competitive and selective peptide-peptoid hybrid (peptomeric) inhibitors might open the way to targeting unwanted proteolysis. PMID- 15883972 TI - Domain-selective ligand-binding modes and atomic level pharmacophore refinement in angiotensin I converting enzyme (ACE) inhibitors. AB - Somatic ACE (EC 3.4.15.1), a Zn(II) metalloproteinase, is composed of functionally active N and C domains resulting from tandem gene duplication. Despite the high degree of sequence similarity between the two domains, they differ in substrate and inhibitor specificity and in their activation by chloride ions. Because of the critical role of ACE in cardiovascular and renal diseases, both domains are attractive targets for drug design. Putative structural models have been generated for the interactions of ACE inhibitors (lisinopril, captoril, enalaprilat, keto-ACE, ramiprilat, quinaprilat, peridoprilat, fosinoprilat, and RXP 407) with both the ACE_C and the ACE_N domains. Inhibitor-domain selectivity was interpreted in terms of residue alterations observed in the four subsites of the binding grooves of the ACE_C/ACE_N domains (S1: V516/N494, V518/T496, S2: F391/Y369, E403/R381, S1': D377/Q355, E162/D140, V379/S357, V380/T358, and S2': D463/E431, T282/S260). The interactions governing the ligand-receptor recognition process in the ACE_C domain are: a salt bridge between D377, E162, and the NH(2) group (P1' position), a hydrogen bond of the inhibitor with Q281, the presence of bulky hydrophobic groups in the P1 and P2' sites, and a stacking interaction of F391 with a benzyl group in the P2 position. In ACE_N these interactions are: hydrogen bonds of the inhibitor with E431, Y369, and R381, and a salt bridge between the carboxy group in the P2 position of the inhibitor and R500. The calculated complexes were evaluated for their consistency with structure-activity relationships and site-directed mutagenesis data. A comparison between the calculated interaction free energies and the experimentally observed biological activities was also made. Pharmacophore refinement was achieved at an atomic level, and might provide an improved basis for structure-based rational design of second-generation, domain-selective inhibitors. PMID- 15883973 TI - Creating space for large secondary alcohols by rational redesign of Candida antarctica lipase B. AB - The active site of Candida antarctica lipase B (CALB) hosts the catalytic triad (Ser-His-Asp), an oxyanion hole and a stereospecificity pocket. During catalysis, the fast-reacting enantiomer of secondary alcohols places its medium-sized substituent in the stereospecificity pocket and its large substituent towards the active-site entrance. The largest group to fit comfortably in the stereospecificity pocket is ethyl, and this restricts the number of secondary alcohols that are good substrates for CALB. In order to overcome this limitation, the size of the stereospecificity pocket was redesigned by changing Trp104. The substrate specificity of the Trp104Ala mutant compared to that of the wild-type lipase increased 270 times towards heptan-4-ol and 5500 times towards nonan-5-ol; this resulted in the high specificity constants 1100 and 830 s(-1) M(-1), respectively. The substrate selectivity changed over 400,000 times for nonan-5-ol over propan-2-ol with both Trp104Ala and the Trp104Gln mutations. PMID- 15883974 TI - Molecular basis of the interaction specificity between the human glucocorticoid receptor and its endogenous steroid ligand cortisol. AB - We analyzed the binding of five steroids to the human glucocorticoid receptor (hGR) experimentally as well as theoretically. In vitro, we measured the binding affinity of aldosterone, cortisol, estradiol, progesterone, and testosterone to hGR in competition with the ligand dexamethasone. The binding affinity relative to the endogenous ligand cortisol (100%) is reduced for progesterone (22%) and aldosterone (20%) and is very weak for testosterone (1.5%) and estradiol (0.2%). In parallel, we constructed a homology model of the hGR ligand-binding domain (LBD) based on the crystal structure of the human progesterone receptor (hPR). After docking the five steroids into the hGR model ligand-binding pocket, we performed five separate 4-ns molecular dynamics (MD) simulations with these complexes in order to study the complex structures. We calculated the binding affinities with two different approaches (MM/PBSA, FlexX) and compared them with the values of the experimentally determined relative binding affinities. Both theoretical methods allowed discrimination between strongly and weakly binding ligands and recognition of cortisol as the endogenous ligand of the hGR in silico. Cortisol binds most strongly due to a nearly perfect steric and electrostatic complementarity with the hGR binding pocket. Chemically similar ligands such as estradiol, testosterone, and progesterone also fit into the hGR binding pocket, but they are unable to form all those contacts with the amino acids of the protein that are necessary to yield a stable, transcriptionally active receptor conformation. Our analysis thus explains the selectivity of the human glucocorticoid receptor for its endogenous ligand cortisol at a molecular level. PMID- 15883975 TI - New inhibitors of the Tat-TAR RNA interaction found with a "fuzzy" pharmacophore model. AB - TAR RNA is a potential target for AIDS therapy. Ligand-based virtual screening was performed to retrieve novel scaffolds for RNA-binding molecules capable of inhibiting the Tat-TAR interaction, which is essential for HIV replication. We used a "fuzzy" pharmacophore approach (SQUID) and an alignment-free pharmacophore method (CATS3D) to carry out virtual screening of a vendor database of small molecules and to perform "scaffold-hopping". A small subset of 19 candidate molecules were experimentally tested for TAR RNA binding in a fluorescence resonance energy transfer (FRET) assay. Both methods retrieved molecules that exhibited activities comparable to those of the reference molecules acetylpromazine and chlorpromazine, with the best molecule showing ten times better binding behavior (IC50 = 46 microM). The hits had molecular scaffolds different from those of the reference molecules. PMID- 15883976 TI - Diatom/copepod interactions in plankton: the indirect chemical defense of unicellular algae. AB - Numerous coexisting species can be observed in the open oceans. This includes the complex community of the plankton, which comprises all free floating organisms in the sea. Traditionally, nutrient limitation, competition, predation, and abiotic factors have been assumed to shape the community structure in this environment. Only in recent years has the idea arisen that chemical signals and chemical defense can influence species interactions in the plankton as well. Key players at the base of the marine food web are diatoms (unicellular algae with silicified cell walls) and their main predators, the herbivorous copepods. It was assumed that diatoms represent a generally good food source for the grazers but recent work indicates that some species use chemical defenses. Secondary metabolites, released by these algae immediately after wounding, are targeted not against the predators themselves but rather at interfering with their reproductive success. This strategy allows diatoms to reduce the grazer population, thereby influencing the marine food web. This review addresses the chemical ecology of the defensive oxylipins formed by diatoms and the question of how these metabolites can act in such a dilute environment. Aspects of biosynthesis, bioassays, and the possible implications of such a chemical defense for the plankton community structure are also discussed. PMID- 15883977 TI - PIGA (N,N-Di-n-butyl-5-chloro-2-(4-chlorophenyl)indol-3-ylglyoxylamide), a new mitochondrial benzodiazepine-receptor ligand, induces apoptosis in C6 glioma cells. AB - Mitochondrial benzodiazepine-receptor (mBzR) ligands constitute a heterogeneous class of compounds that show a pleiotropic spectrum of effects within the cells, including the modulation of apoptosis. In this paper, a novel synthetic 2 phenylindol-3-ylglyoxylamide derivative, N,N-di-n-butyl-5-chloro-2-(4 chlorophenyl)indol-3-ylglyoxylamide (PIGA), which shows high affinity and selectivity for the mBzR, is demonstrated to induce apoptosis in rat C6 glioma cells. PIGA was able to dissipate mitochondrial transmembrane potential (DeltaPsim) and to cause a significant cytosolic accumulation of cytochrome c. Moreover, typical features of apoptotic cell death, such as caspase-3 activation and DNA fragmentation, were also detected in PIGA-treated cells. Our data expand the knowledge on mBzR ligand-mediated apoptosis and suggest PIGA as a novel proapoptotic compound with therapeutic potential against glial tumours, in which apoptosis resistance has been reported to be involved in carcinogenesis. PMID- 15883978 TI - Trapping of proteins under physiological conditions in a nanopipette. PMID- 15883979 TI - Kanamycin A-derived cationic lipids as vectors for gene transfection. AB - Cationic lipids nowadays constitute a promising alternative to recombinant viruses for gene transfer. We have recently explored the transfection potential of a new class of lipids based upon the use of aminoglycosides as cationic polar headgroups. The encouraging results obtained with a first cholesterol derivative of kanamycin A prompted us to investigate this family of vectors further, by modulating the constituent structural units of the cationic lipid. For this study, we have investigated the transfection properties of a series of new derivatives based on a kanamycin A scaffold. The results primarily confirm that aminoglycoside-based lipids are efficient vectors for gene transfection both in vitro and in vivo (mouse airways). Furthermore, a combination of transfection and physicochemical data revealed that some modifications of the constitutive subunits of kanamycin A-based vectors were associated with substantial changes in their transfection properties. PMID- 15883980 TI - Prevalence of risk factors for suicide in patients prescribed venlafaxine, fluoxetine, and citalopram. AB - PURPOSE: Three recent observational studies reported that the risk of fatal overdose is greater with venlafaxine than SSRI use. It is not clear whether patient factors could account for this finding. We evaluated whether risk factors for suicide are more prevalent among patients prescribed venlafaxine than patients prescribed fluoxetine or citalopram. METHODS: Using data from the UK General Practice Research Database (GPRD), we identified patients who were first prescribed any of the three drugs between January 1995 and April 2002. We ascertained risk factors for suicide documented in the 1 year before that first prescription. Separate analyses compared venlafaxine (N = 27 096) and fluoxetine (N = 134 996) cohorts, and venlafaxine and citalopram (N = 52 035) cohorts. RESULTS: Previous suicidal behaviors were documented for 1.0% of the venlafaxine cohort compared to 0.4% of the fluoxetine cohort (OR 2.8, 95%CI: 2.4, 3.2) and 0.4% citalopram cohorts (OR 2.4, 95%CI: 2.0, 2.9). 72.5% of venlafaxine patients had been prescribed at least one other antidepressant compared to 27.6% of fluoxetine (OR 6.9, 95%CI: 6.7, 7.1) and 39.5% of citalopram (OR 4.0, 95%CI: 3.9, 4.2) patients. Venlafaxine patients were also four to six times as likely to have been previously hospitalized for depression. CONCLUSION: In the UK, venlafaxine has been selectively prescribed to a patient population with a higher burden of suicide risk factors than patients prescribed fluoxetine and citalopram. Unless baseline population differences are accounted for, observational studies that compare the risk of suicide in patients receiving these agents may produce biased results. PMID- 15883981 TI - Theoretical and experimental NMR study of protopine hydrochloride isomers. AB - The 1H and 13C NMR chemical shifts of cis- and trans-protopinium salts were measured and calculated. The calculations of the chemical shifts consisted of conformational analysis, geometry optimization (RHF/6-31G** method) and shielding constants calculations (B3LYP/6-31G** method). Based on the results of the quantum chemical calculations, two sets of experimental chemical shifts were assigned to the particular isomers. According to the experimental results, the trans-isomer is more stable and its population is approximately 68%. PMID- 15883982 TI - Biometry of the pubovisceral muscle and levator hiatus by three-dimensional pelvic floor ultrasound. AB - OBJECTIVE: Until recently, magnetic resonance was the only imaging method capable of assessing the levator ani in vivo. Three-dimensional (3D) ultrasound has recently been shown to be able to demonstrate the pubovisceral muscle. The aim of this study was to define the anatomy of the levator hiatus in young nulliparous women with the help of 3D ultrasound. METHODS: In a prospective observational study, 52 nulligravid female Caucasian volunteers (aged 18-24 years) were assessed by two-dimensional (2D) and 3D translabial ultrasound after voiding whilst supine. Pelvic organ descent was assessed on Valsalva maneuver. Volumes were acquired at rest and on Valsalva maneuver, and biometric indices of the pubovisceral muscle and levator hiatus were determined in the axial and coronal planes. RESULTS: In the axial plane, average diameters of the pubovisceral muscle were 0.4-1.1 cm (mean 0.73 cm). Average area measurements were 7.59 (range, 3.96 11.9) cm2. The levator hiatus at rest varied from 3.26 to 5.84 (mean 4.5) cm in the sagittal direction, and from 2.76 to 4.8 (mean 3.75) cm in the coronal plane. The hiatus area at rest ranged from 6.34 to 18.06 (mean 11.25) cm2 increasing to 14.05 (6.67-35.01) cm(2) on Valsalva maneuver (P = 0.009). There were significant correlations between pelvic organ mobility and hiatus area at rest (P = 0.018 to P < 0.001) and on Valsalva maneuver (all P < 0.001). CONCLUSIONS: Biometric indices of the pubovisceral muscle and levator hiatus can be determined by 3D ultrasound. Significant correlations exist between hiatal area and pelvic organ descent. These data provide support for the hypothesis that levator ani anatomy plays an independent role in determining pelvic organ support. PMID- 15883983 TI - Reference ranges for serial measurements of blood velocity and pulsatility index at the intra-abdominal portion, and fetal and placental ends of the umbilical artery. AB - OBJECTIVE: To construct reference ranges for serial measurements of umbilical artery (UA) blood flow velocity and pulsatility index (PI) at standardized insonation sites during the second half of pregnancy. METHODS: This was a prospective longitudinal study of the umbilical circulation. UA blood flow velocities were measured at the intra-abdominal portion, fetal end and placental end at 4-weekly intervals at 19-42 weeks of gestation in 130 low-risk singleton pregnancies. A total of 513 observations were used to construct the reference ranges using regression statistics and multilevel modeling. RESULTS: UA blood velocities and PI were higher at the intra-abdominal portion and fetal end than at the placental end. The gestational age-related increase of end-diastolic velocity was greater than the corresponding increase of the peak systolic velocity at all locations. The mean differences (delta values) of UA blood velocities between the fetal and placental ends increased and that of PI decreased with advancing gestational age. CONCLUSION: UA Doppler parameters vary significantly at different locations. We have established new reference ranges for the UA velocities and PI at standardized locations based on longitudinal observations, which should be useful for the surveillance of fetuses with repeated observations. PMID- 15883984 TI - Synthesis and properties of chiral ammonium-based ionic liquids. AB - New chiral ammonium-based ionic liquids containing the (1R,2S,5R)-(-)-menthyl group can be easily and efficiently prepared under ambient conditions. The preparation and characterization of trialkyl[(1R,2S,5R)-(-) menthoxymethyl]ammonium salts is reported. The salts have been demonstrated to be air- and moisture-stable under ambient conditions and can be readily used in a variety of standard experimental procedures. The single-crystal X-ray structure of butyldimethyl[(1R,2S,5R)-(-)-menthoxymethyl]ammonium chloride has been determined. The chiral, room-temperature ionic liquids have been characterized by physical properties such as specific rotation, density, viscosity, thermal degradation, and glass transition temperature. Trialkyl[(1R,2S,5R)-(-) menthoxymethyl]ammonium chloride prototype ionic liquids have also been found to exhibit strong antimicrobial and high antielectrostatic activities. PMID- 15883985 TI - Effect of axial ligation or pi-pi-type interactions on photochemical charge stabilization in "two-point" bound supramolecular porphyrin-fullerene conjugates. AB - Two types of structurally well-defined, self-assembled zinc porphyrin-fullerene conjugates were formed by "two-point" binding strategies to probe the effect of axial ligation or pi-pi-type interactions on the photochemical charge stabilization in the supramolecular dyads. To achieve this, meso tetraphenylporphyrin was functionalized to possess one or four [18]crown-6 moieties at different locations on the porphyrin macrocycle while fullerene was functionalized to possess an alkyl ammonium cation, and a pyridine or phenyl entities. As a result of the crown ether-ammonium cation complexation, and zinc pyridine coordination or pi-pi-type interactions, stable zinc porphyrin-fullerene conjugates with defined distance and orientation were formed. Evidence for the zinc-pyridine complexation or pi-pi-type interactions was obtained from the spectral and computational studies. Steady-state and time-resolved emission studies revealed efficient quenching of the zinc-porphyrin singlet excited state in these dyads, and the measured rates of charge separation, k(CS) were found to be slightly better in the case of the dyads held by axial coordination and crown ether-cation complexation. Nanosecond transient absorption studies provided evidence for the electron transfer reactions, and these studies also revealed charge stabilization in these dyads. The lifetimes of the radical ion pairs were found to depend upon the type of porphyrins utilized to form the dyads, that is, porphyrin possessing the crown ether moiety at the ortho position of one of the phenyl rings yielded prolonged charge stabilized states. Addition of pyridine to the supramolecular dyads eliminated the zinc-pyridine coordination or pi-pi-type interactions of the "two-point" bound systems due to the formation of a new zinc pyridine axial bond thus giving a unique opportunity to probe the effect of axial coordination or pi-pi interactions on k(CS) and k(CR). Under these conditions, the measured electron transfer rates revealed faster k(CS) and slower k(CR) as compared to those obtained in the absence of added pyridine. The evaluated lifetimes of the radical ion-pairs were found to be hundreds of nanoseconds and were longer in the presence of pyridine. PMID- 15883986 TI - Microheterogeneous solvation for aminolysis reactions in AOT-based water-in-oil microemulsions. AB - A kinetic study was carried out on the aminolysis of p-nitrophenyl acetate (NPA) by n-decylamine (DEC), piperazine (PIP) and sarcosine (SAR) in AOT/isooctane/water (w/o) microemulsions. By using the pseudophase model both the rate constants at the interface, k2i, and the water microdroplet, k2w, can be obtained. The obtained results show that k2i increases together with the water content of the microemulsion, whereas k2w increases as the water content of the system decreases. In the aqueous microdroplet the predominant interaction Na+...OH2 causes a decrease in the strength of the hydrogen bonds and therefore facilitates the desolvation of the reagents as W decreases. This desolvation of the reagents causes the increase of k2w as W decreases. In the interface of the microemulsion the predominant interaction SO3-...HOH causes an increase in the electronic density on the water molecules and the consequent decrease in their efficiency in the solvation of the partial negative charge, which develops on the carbonyl oxygen atom in the transition state of the reaction. This decrease in the solvation causes k2i to decrease together with the water content of the system. PMID- 15883987 TI - Highly hydrated cations: deficiency, mobility, and coordination of water in crystalline nonahydrated scandium(III), yttrium(III), and lanthanoid(III) trifluoromethanesulfonates. AB - Trivalent lanthanide-like metal ions coordinate nine water oxygen atoms, which form a tricapped trigonal prism in a large number of crystalline hydrates. Water deficiency, randomly distributed over the capping positions, was found for the smallest metal ions in the isomorphous nonahydrated trifluoromethanesulfonates, [M(H2O)n](CF3SO3)3, in which M = Sc(III), Lu(III), Yb(III), Tm(III) or Er(III). The hydration number n increases (n = 8.0(1), 8.4(1), 8.7(1), 8.8(1) and 8.96(5), respectively) with increasing ionic size. Deuterium (2H) solid-state NMR spectroscopy revealed fast positional exchange between the coordinated capping and prism water molecules; this exchange started at temperatures higher than about 280 K for lutetium(III) and below 268 K for scandium(III). Similar positional exchange for the fully nonahydrated yttrium(III) and lanthanum(III) compounds started at higher temperatures, over about 330 and 360 K, respectively. An exchange mechanism is proposed that can exchange equatorial and capping water molecules within the restrictions of the crystal lattice, even for fully hydrated lanthanoid(III) ions. Phase transitions occurred for all the water-deficient compounds at approximately 185 K. The hydrated scandium(III) trifluoromethanesulfonate transforms reversibly (DeltaH degrees = -0.80(1) kJ mol(-1) on cooling) to a trigonal unit cell that is almost nine times larger, with the scandium ion surrounded by seven fully occupied and two partly occupied oxygen atom positions in a distorted capped trigonal prism. The hydrogen bonding to the trifluoromethanesulfonate anions stabilises the trigonal prism of water ligands, even for the crowded hydration sphere of the smallest metal ions in the series. Implications for the Lewis acid catalytic activity of the hydrated scandium(III) and lanthanoid(III) trifluoromethanesulfonates for organic syntheses performed in aqueous media are discussed. PMID- 15883988 TI - Optimal spike coherence and synchronization on complex Hodgkin-Huxley neuron networks. PMID- 15883989 TI - Lewis acidity/basicity of pi-electron systems: theoretical study of a molecular interaction between a pi system and a Lewis acid/base. AB - Molecular interactions between pi systems having different pi-electron character (benzene, hexafluorobenzene, and borazine), and a Lewis acid/base (borane and ammonia) were theoretically studied. An attractive interaction between benzene, the electron-rich pi system, and borane was observed. On the other hand, repulsive interactions between benzene and ammonia was observed when the lone pair of nitrogen points toward the benzene ring. In contrast, an attractive interaction between hexafluorobenzene, an electron-deficient pi system, and ammonia was observed. Unexpectedly, a weak attractive interaction between hexafluorobenzene and borane was also observed. Borazine shows an interaction both to borane and ammonia. The attraction between the nitrogen atom of borazine and borane was larger than that between the boron atom of borazine and ammonia. PMID- 15883990 TI - Weak backscattering in deflected doped carbon nanotubes. PMID- 15883991 TI - Particle distribution in a microporous material: experiments with zeolite A. AB - The theory on particle distribution and exchange equilibria in a microporous material is applied to experimental ion-exchange data involving zeolite Na-A and zeolite K-A, with silver ions as the exchanging species. The presented method enables direct evaluation of the measured data and consideration of nonequivalent particle sites. The isotherms of the K+ versus Ag+ exchange in zeolite K-A rise much more steeply, at low exchange degrees, than those of the Na+ versus Ag+ exchange in zeolite Na-A. This result implies a different course of the ion exchange reactions. Spectroscopic measurements on dehydrated, partly silver exchanged zeolites Na-A and K-A do indeed show that in zeolite Na-A, the Na+ ions occupying four-ring positions are exchanged faster for Ag+ than the Na+ ions occupying eight- and six-ring positions, while in zeolite K-A the exchange does not start with the four-ring ion but with six-ring ions, followed by the four ring ion. These findings are consistent with the results obtained from evaluation of the ion-exchange data. The resulting thermodynamic quantities significantly differ from published reference values, which we suggest should be revised. PMID- 15883992 TI - Surface-structure-sensitive adsorption of adenine on gold electrodes. PMID- 15883993 TI - Switching mechanism in polar columnar mesophases made of bent-core molecules. AB - The behaviour of polar, broken-layer-type columnar phases made of bent molecules (B(1Rev) and B(1RevTilted)) was studied under an applied electric field. There are two competing mechanisms of ferroelectric switching in the polar B(1RevTilted) columnar phase: collective rotation around the long molecular axis and collective rotation around the tilt cone. The proposed model shows that the main factor discriminating the type of switching is the width of the column cross section. PMID- 15883994 TI - A computational study of thiolate and selenolate oxidation by hydrogen peroxide. AB - Ab initio molecular orbital calculations have been used to study the effects of the molecular environment on the oxidation of thiolate and selenolate by hydrogen peroxide. The reaction was first examined in vacuo at the QCISD(T)/6 311+G(2df,2pd)//MP2/6-311+G(d,p) level of theory. It was found for both thiolate and selenolate that a reactant aggregate is formed, which has a dissociation rate constant comparable to the activation rate constant (about 10(-3) s(-1) for thiolate and 10(-1) s(-1) for selenolate). Using the polarizable continuum model (PCM) it was then found that the dissociation barrier energy decreases dramatically in water giving a dissociation rate constant of the order of 10(9) s(-1). In this case, the predicted overall rate constant of the thiolate reaction was about 10.2 mol(-1) dm3 s(-1), which is in good agreement with the experimental rate constant of cysteine oxidation in aqueous solution. The calculated rate constant for the selenolate reaction was somewhat higher (about 35.4 mol(-1) dm3 s(-1)). However, this value is several orders of magnitude smaller than the experimental value reported for the oxidation of selenocysteine in glutathione peroxidase. By considering the effect of the PCM dielectric constant on the reaction rate constant it was concluded that the high reactivity of the selenocysteine in glutathione peroxidase, as compared with cysteine, could be mainly due to the molecular environment of the selenocysteine residue. PMID- 15883995 TI - Mesoscopic structures of maghemite nanocrystals: fabrication, magnetic properties, and uses. AB - Mesoscopic structures made of maghemite (gamma-Fe2O3) nanocrystals with different coatings and shapes are described. They are controlled by a combination of van der Waals and magnetic dipolar interactions and their shapes are responsible for the collective magnetic response of the nanocrystals. It is shown that spherical gamma-Fe2O3 nanocrystals can be used as a mask to reproduce the mesoscopic structure on a silicon wafer. PMID- 15883996 TI - Stepwise shrinkage of liposomes driven by thermal fluctuations of the membranes. PMID- 15883997 TI - Probing binding-mode diversity in guanidinium-oxoanion host-guest systems. AB - An attempt to experimentally estimate the role of binding-mode diversity (structural fuzziness) on the molecular recognition seen in the prominent guanidinium-oxoanion host-guest pair is described. The global heat response as measured by isothermal titration calorimetry in acetonitrile, which was obtained from the interaction of five different but structurally closely related guanidinium hosts with three rigid phosphinate guests of decreasing accessibility of their binding sites, is correlated to provide a trend analysis. All host-guest associations of 1:1 stoichiometry in this series are strongly enthalpy-driven. The change in complexation entropy can be related to the tightness of the mutual fit of the host-guest partners, which approaches a minimum limit and is interpreted as the unique lock-and-key binding mode. The ordinary host-guest complexation in this ensemble features substantial positive entropy changes that correlate inversely with the binding interface area. This finding excludes desolvation effects as the major cause of entropy production, and provides evidence for the existence of a broad variety of complex configurations rather than a single binding mode to represent the associated host-guest pair. This result bears on the molecular design of systems that vitally depend on structural fidelity, such as nanoassemblies or homogeneous catalysis. PMID- 15883998 TI - The influence of solid-state molecular organization on the reaction paths of thiyl radicals. AB - Electron paramagnetic resonance (EPR) spectroscopy has been employed to investigate the effect of solid-state molecular organization on the reaction of thiyl radicals with thiols. In an irradiated C18H37SH/thiourea clathrate, the conversion of thiyl to perthiyl radicals is substantial, due to the head-to-head arrangement of the reactants within the channels and the suppression of other possible competing reactions due to hindrance by the clathrate walls. The perthiyl radical was identified using EPR analysis of its molecular dynamics within the clathrate channels. Irradiated polyethylene film containing 30% C18H37SH afforded a negligible conversion of thiyl to perthiyl radicals because of the random distribution of reactants. These results suggest that in the absence of favorable structure-control effects, the reaction between RS* and RSH is unimportant with respect to other competing reactions. Perthiyl radicals are also the major product in the vacuum solid-state radiolysis of lysozyme. A proposal of the mechanism involved in all cases is based on the equilibrium RS* + RSH <==> RSS*(H)R, followed by the irreversible conversion of the sulfuranyl radical to the perthiyl radical. As the equilibrium is strongly shifted to the left, the intermediate sulfuranyl radicals were not detected, but the lack of other competing reactions for the thiyl radicals caused the formation of perthiyl radicals to become the major path in the clathrate and in solid lysozyme radiolysis. PMID- 15883999 TI - Local translational control in dendrites and its role in long-term synaptic plasticity. AB - Local protein synthesis in dendrites has emerged as a key mechanism contributing to enduring forms of synaptic plasticity. Although the translational capability of dendrites has been appreciated for over 20 years, it is only recently that significant progress has been made in elucidating mechanisms that contribute to its regulation. It is clear from work over the last few years that the control of translation in dendrites is complex, involving a host of unique (and often surprising) mechanisms that can operate together or in parallel to tightly control gene expression in time and space. Here, we discuss the strategies used by neurons to regulate translation in dendrites and how these are implemented in the service of long-term information storage. PMID- 15884000 TI - Computational modeling of dendrites. AB - Computational methods have been part of neuroscience for many years. For example, models developed with these methods have provided a theory that helps explain the action potential. More recently, as experimental patch-electrode techniques have revealed new biophysics related to dendritic function and synaptic integration, computational models of dendrites have been developed to explain and further illuminate these results, and to predict possible additional behavior. Here, a collection of computational models of dendrites is reviewed. The goal is to help explain how such computational techniques work, some of their limitations, and what one can hope to learn about dendrites by modeling them. PMID- 15884001 TI - Plasticity of dendritic excitability. AB - Dendrites are equipped with a plethora of voltage-gated ion channels that greatly enrich the computational and storage capacity of neurons. The excitability of dendrites and dendritic function display plasticity under diverse circumstances such as neuromodulation, adaptation, learning and memory, trauma, or disorders. This adaptability arises from alterations in the biophysical properties or the expression levels of voltage-gated ion channels-induced by the activity of neurotransmitters, neuromodulators, and second-messenger cascades. In this review we discuss how this plasticity of dendritic excitability could alter information transfer and processing within dendrites, neurons, and neural networks under physiological and pathological conditions. PMID- 15884002 TI - Rho GTPases, dendritic structure, and mental retardation. AB - A consistent feature of neurons in patients with mental retardation is abnormal dendritic structure and/or alterations in dendritic spine morphology. Deficits in the regulation of the dendritic cytoskeleton affect both the structure and function of dendrites and synapses and are believed to underlie mental retardation in some instances. In support of this, there is good evidence that alterations in signaling pathways involving the Rho family of small GTPases, key regulators of the actin and microtubule cytoskeletons, contribute to both syndromic and nonsyndromic mental retardation disorders. Because the Rho GTPases have been shown to play increasingly well-defined roles in determining dendrite and dendritic spine development and morphology, Rho signaling has been suggested to be important for normal cognition. The purpose of this review is to summarize recent data on the Rho GTPases pertaining to dendrite and dendritic spine morphogenesis, as well as to highlight their involvement in mental retardation resulting from a variety of genetic mutations within regulators and effectors of these molecules. PMID- 15884003 TI - Synaptic integration in dendritic trees. AB - Most neurons have elaborate dendritic trees that receive tens of thousands of synaptic inputs. Because postsynaptic responses to individual synaptic events are usually small and transient, the integration of many synaptic responses is needed to depolarize most neurons to action potential threshold. Over the past decade, advances in electrical and optical recording techniques have led to new insights into how synaptic responses propagate and interact within dendritic trees. In addition to their passive electrical and morphological properties, dendrites express active conductances that shape individual synaptic responses and influence synaptic integration locally within dendrites. Dendritic voltage-gated Na(+) and Ca(2+) channels support action potential backpropagation into the dendritic tree and local initiation of dendritic spikes, whereas K(+) conductances act to dampen dendritic excitability. While all dendrites investigated to date express active conductances, different neuronal types show specific patterns of dendritic channel expression leading to cell-specific differences in the way synaptic responses are integrated within dendritic trees. This review explores the way active and passive dendritic properties shape synaptic responses in the dendrites of central neurons, and emphasizes their role in synaptic integration. PMID- 15884004 TI - Gonadal hormone modulation of dendrites in the mammalian CNS. AB - This review focuses on the effect of gonadal steroid hormones, androgen and estrogen, on dendrites in the adult rat central nervous system (CNS). Four hormone-responsive nuclei are considered: The spinal nucleus of the bulbocavernosus (SNB), the medial nucleus of the amygdala (MeA), the ventromedial nucleus of the hypothalamus (VMN), and the CA1 region of the dorsal hippocampus. Particular emphasis is placed on the mode of hormone action in each nucleus. In the SNB, VMN, and hippocampus, hormones appear to mediate their effects indirectly, via cells other than those that display morphological plasticity. In the MeA, estrogen and/or androgen appears to act primarily on those cells whose dendrites are modulated by the hormone. Importantly, increasing levels of gonadal hormones do not simply result in increases in dendritic parameters. In the VMN, high levels of estrogen associated with proestrus increase dendritic spine density in one subset of cells and reduce spine density in another subset. The pyramidal cells of dorsal CA1 also undergo phasic changes in dendritic spine and synapse density across the estrous cycle. The estrogen-induced excitatory synapses connect with preexisting axonal boutons that also form synapses with other CA1 cells, thereby increasing the divergence of excitatory afferents to dorsal CA1. These findings indicate that gonadal steroids have a profound impact on the morphology of dendrites and patterns of synaptic connectivity. Consequently, the experimental manipulation of hormone levels is a powerful tool to study structure-function relationships in the mammalian brain. PMID- 15884005 TI - Dendritic spine morphogenesis and plasticity. AB - Dendritic spines are small protrusions off the dendrite that receive excitatory synaptic input. Spines vary in size, likely correlating with the strength of the synapses they form. In the developing brain, spines show highly dynamic behavior thought to facilitate the formation of new synaptic contacts. Recent studies have illuminated the numerous molecules regulating spine development, many of which converge on the regulation of actin filaments. In addition, interactions with glial cells are emerging as important regulators of spine morphology. In many cases, spine morphogenesis, plasticity, and maintenance also depend on synaptic activity, as shown by recent studies demonstrating changes in spine dynamics and maintenance with altered sensory experience. PMID- 15884008 TI - Maturation of dendritic architecture: lessons from insect identified neurons. AB - The highly complex geometry of dendritic trees is crucial for neural signal integration and the proper wiring of neuronal circuits. The morphogenesis of dendritic trees is regulated by innate genetic factors, neuronal activity, and external molecular cues. How each of these factors contributes to dendritic maturation has been addressed in studies of the developing nervous systems of animals ranging from insects to mammals. This article reviews our current knowledge and understanding of the role of afferent input in the establishment of the architecture of mature dendritic trees, using insect neurons as models. With these model systems and using quantitative morphometry, it is possible to define the contributions of intrinsic and extrinsic factors in dendritic morphogenesis of identified neurons and to evaluate the impact of dendritic maturation on the integration of identified neurons into functional circuits subserving identified behaviors. The commonly held view of dendritic morphogenesis is that general structural features result from genetic instructions, whereas fine connectivity details rely mostly on substrate interactions and functional activity. During early dendritic maturation, dendritic growth cone formation produces new branches at all dendritic roots. The second phase is growth cone independent and afferent input dependent, during which branching is limited to high order distal dendrites. During the third phase, activity-dependent synaptic maturation occurs with limited or subtle remodeling of branching. PMID- 15884009 TI - Remodeling dendrites during insect metamorphosis. AB - The genesis of dendritic shape during development sets in place key characteristics of a neuron's physiology and connectivity. During this construction, a cell interprets intrinsic cell-specific developmental programs and cues from the environment to generate its final phenotype. In insects that undergo complete metamorphosis certain neurons function in the larval nervous system and then remodel to generate an adult-specific arbor. By studying the dendrites of neurons that undergo such a cellular metamorphosis, one can explore the mechanisms that underlie both stereotyped pruning and local remodeling. Live imaging techniques in intact Drosophila have been especially useful in examining the outgrowth of the adult-specific dendritic arbors in remodeling dendritic arborizing (da) sensory neurons. These neurons show an initial scaffold-building phase during which the cell establishes the overall shape of the arbor and then switch to an elaboration phase where the arbor is filled out with higher order branches. The cellular machinery employed during these two phases is different, with branch retraction being a prominent feature of the scaffold building phase but absent from the elaboration phase. The transition between these two modes does not appear to be "hard-wired" but is plastic and under the extrinsic control of developmental hormones. This transition in branch dynamics may also involve changes in calcium signaling in the growing arbor. The potential relationship between hormone-induced transcriptional change and the calcium dynamics in dendritic morphogenesis is discussed. PMID- 15884010 TI - Regulation of dendritic development by neuronal activity. AB - Proper development of dendrites is essential for the establishment of neuronal circuitry. The elaboration of the dendritic tree is a highly dynamic and regulated process, which involves the formation of new branches as well as the maintenance or elimination of pre-existing branches. This review describes recent advances in our understanding of the molecular mechanisms of activity-dependent dendritic development. Neuronal activity triggers calcium-mediated signaling events that affect the structural components of dendrites and adhesion molecules. These calcium-induced signaling pathways also target nuclear transcription factors thereby controlling expression of genes required for dendritic development. Thus, a coordinated response to calcium-regulated signaling pathways mediates activity-dependent dendritic development. PMID- 15884011 TI - Unilateral acheiria and fatal primary pulmonary hypertension in a girl with incontinentia pigmenti. AB - We describe a newborn girl with incontinentia pigmenti (IP, MIM308300), unilateral acheiria, and fatal primary pulmonary hypertension. Limb deficiency has not been described previously in IP and pulmonary hypertension only on two previous occasions. A review of the cause of IP shows that these rare manifestations may not be unexpected, given the many roles of the underlying gene product. PMID- 15884012 TI - Moderate lead exposure elicits neurotrophic effects in cerebral cortical precursor cells in culture. AB - Lead (Pb) persists as an environmental toxicant despite aggressive environmental and occupational regulation. Neurotoxicological effects of acute Pb poisoning range from subtle cognitive deficits, to clumsiness and ataxia, to coma and seizures. In adult neurotoxicity, reductions of blood Pb levels are often associated with reversal of clinical signs. In children, however, the effects are more likely to endure, with even low levels of chronic Pb exposure correlating with decreasing IQ. These persistent effects likely result from neurodevelopmental insults, such as altered cell survival or maturation, although the mechanisms have not been fully defined. In the present study we define the effects of moderate-level Pb exposure on mammalian neurogenesis using a well characterized cortical precursor model. Gestational day 14.5 rat cerebral cortical precursor cells were cultured in defined media and cell number, precursor proliferation, apoptosis, and neuritic process outgrowth were assessed following exposure to a range of Pb acetate concentrations. Surprisingly, whereas a concentration of 30 microg/ml Pb acetate was acutely toxic to neurons, concentrations between 1 and 10 microg/ml Pb acetate (approximately 3 microM and 30 microM Pb, respectively) increased cell number: 10 times as many cells exposed to 10 microg/ml Pb were present on day 4 as compared to control. The increase in cell number was not a result of increased proliferation, however, as DNA synthesis did not increase. Rather, Pb exposure maintained the survival of cortical precursors, as the progressive apoptosis occurring under control conditions was markedly reduced by the metal. Additionally, neuritic process initiation and outgrowth increased in a concentration-dependent manner, with processes four times as abundant on day 1 and twice as long on day 2. These results suggest that brief exposure to lead during neurogenesis directly affects cell survival and process development, potentially altering cortical arrangement. Consequently, alterations in neural circuitry may underlie some of the neurological effects of Pb exposure during brain development. PMID- 15884014 TI - Neural cell adhesion molecule function is regulated by metalloproteinase-mediated ectodomain release. AB - The neural cell adhesion molecule (NCAM) is involved in development of the nervous system, in brain plasticity associated with learning and memory, and in neuronal regeneration. NCAM regulates these processes by influencing cell adhesion, cell migration, and neurite outgrowth. NCAM activates intracellular signaling upon homophilic NCAM binding, and this is a prerequisite for NCAM stimulated neurite outgrowth. NCAM is synthesized in three main membrane-bound isoforms, NCAM-120, NCAM-140, and NCAM-180. Soluble forms of NCAM in blood and cerebrospinal fluid have also been found, although the functional significance of these forms remains unclear. In this report, we demonstrate that NCAM can be released from primary hippocampal neurons in culture. The release was enhanced by application of ATP and inhibited by the metalloproteinase inhibitor BB-3103. ATP also induced metalloproteinase-dependent release of all three major NCAM isoforms from NCAM-transfected fibroblastoid L-cells. In this model system, the extracellular ATP-binding site of NCAM was shown not to be necessary for ATP induced NCAM release. Furthermore, inhibition of serine, cysteine, and aspartic proteinases could not prevent ATP-induced down-regulation of NCAM in L-cells, suggesting that NCAM is cleaved directly by a metalloproteinase. Aggregation of hippocampal neurons in culture was increased in the presence of the metalloproteinase inhibitor GM 6001, consistent with a metalloproteinase dependent shedding of NCAM occurring in these cells. Moreover, NCAM-dependent neurite outgrowth was significantly reduced by application of GM 6001. Taken together, these results suggest that membrane-bound NCAM can be cleaved extracellularly by a metalloproteinase and that metalloproteinase-dependent shedding of NCAM regulates NCAM-mediated neurite outgrowth. PMID- 15884015 TI - Microglia-derived tumor necrosis factor-alpha exaggerates death of newborn hippocampal progenitor cells in vitro. AB - Production of new hippocampal neurons continues in adult mammals and different brain insults can significantly increase this process. However, many hippocampal progenitor cells (HPC) die shortly after birth. Here we investigated the possibility that increased release of cytokines by activated microglia contributes to the death of HPC. We showed that addition of tumor necrosis factor alpha (TNFalpha) to the medium of a cultured HPC line (HiB5) shortly after the cells stopped division causes significant apoptotic cell death. Conditioned medium from an activated microglial cell line (BV-2) had a similar effect, though conditioned medium from nonactivated microglia increased the survival of HPC. Reverse transcription-PCR indicated that HPC and microglial cells express both TNF receptors, TNF-R1 and TNF-R2. Coculturing of HPC with activated microglial cells aggravated death of hippocampal progenitors and also caused death of microglial cells themselves. Our data indicate that activated microglia-released TNFalpha might be an important contributor in inflammation-induced exaggeration of death of newly formed HPC in the adult brain after an insult. PMID- 15884016 TI - MAPKs are differentially modulated in arctic ground squirrels during hibernation. AB - Hibernating animals are very tolerant of trauma to the central nervous system such that dramatic fluctuations in cerebral blood flow occur during hibernation and arousal without apparent damage. Indeed, it was demonstrated that Arctic ground squirrels (AGS) experience acute and severe systemic hypoxia along with the dramatic fluctuation in cerebral blood flow when the animals are aroused from hibernation. While initial hypotheses concerned protective mechanisms in the hibernating state, recent evidence of sustained elevation of HIF1alpha in euthermic AGS from our laboratory suggests that a preparatory program of protective gene expression is chronically expressed in euthermic AGS. In this study we evaluated potential neuroprotective adaptations by examining the alteration of intracellular MAPK pathways that may be modulated by hypoperfusion/reperfusion in AGS during hibernation and arousal. We found that ERK and JNK are activated in both euthermic and aroused AGS compared to the hibernating group which positively correlated with HIF1alpha levels. The activation of ERK and JNK associated with HIF1alpha may play an important role in mediating neuroprotective adaptations that is essential for successful hibernation. Interestingly, p38 is activated in euthermic AGS but not in aroused AGS, which shows strong correlation with iNOS induction. Therefore, the attenuation of p38 activation and iNOS induction in hibernating and aroused animals may contribute to the attenuation of inflammation that plays important neuroprotective roles during hibernation. Taken together, the differential modulation of the MAPK pathways may be critical for neuroprotection of AGS necessary for fluctuations in oxygen and nutrient delivery during hibernation. PMID- 15884017 TI - Variable alternative spliced exon (VASE)-containing and VASE-lacking neural cell adhesion molecule in the dorsal and ventral hippocampus of SAMP8 mice. AB - The neural cell adhesion molecule (NCAM) is involved in the development and synaptic plasticity of the brain. Differential splicing of the variable alternative spliced exon (VASE) in the fourth immunoglobulin domain can dramatically change the functional properties of NCAM. This paper discusses our analysis of the levels of different expression of VASE-containing NCAM (NCAM VASE(+)) and VASE-lacking NCAM (NCAM-VASE(-)) mRNAs in the dorsal and ventral hippocampus of senescence-accelerated mice (SAM). We further investigated the individual level of NCAM-VASE(+) and NCAM-VASE(-) in relation to the capacity for spatial learning and memory as assessed by a Morris water maze task. The results showed that the levels of both NCAM-VASE(+) and NCAM-VASE(-) were increased significantly in dorsal but not ventral hippocampus in aged SAMP8 mice. The mean latencies to find the hidden platform of the learning task on the last day were positively correlated with the levels of NCAM-VASE(+) in the dorsal hippocampus of SAMP8, which reveals that the mice with high levels of NCAM-VASE(+) have poor learning performances. These results suggest that the up-regulation of NCAM VASE(+) could be involved in the impairments of spatial learning and memory. PMID- 15884018 TI - A new DAX1 gene mutation associated with congenital adrenal hypoplasia and hypogonadotropic hypogonadism. AB - We report on a DAX1 gene investigation in a patient with X-linked adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HH) in order to identify mutations causing this disorder and to confirm the clinical diagnosis. The description of the clinical course of the condition with a detailed documentation of longitudinal data is also reported. A male newborn was referred at 45 days of life because of vomiting, dehydration, and weight loss. The diagnosis was primary adrenal insufficiency. The appropriateness of glucocorticoid therapy during the prepubertal period was difficult to judge because of elevated ACTH levels on one hand and progressive retardation of bone age on the other hand. Basal and GnRH stimulated gonadotropin levels remained low during the entire period of examination and exogenous gonadotropin treatment was begun. This had to be interrupted at age 14.6 years because of the occurrence of a 3rd degree anaplastic ependimoma of the left posterior-parietal region, without apparent lepto-meningeal involvement. The molecular analysis of DAX1 gene of the propositus showed deletion of nucleotides AAT in exon 2, resulting in the loss of the Asn430. No alterations were found in the mother and grandmother. This deleted residue lies in one of the helices forming the hydrophobic core of the ligand binding domain (LBD); thus this mutation may be the cause of the observed phenotype. Further investigations are needed to verify its causal role in AHC associated with HH. PMID- 15884019 TI - Hypoglycemia enhances the expression of prion protein and heat-shock protein 70 in a mouse neuroblastoma cell line. AB - Cellular prion protein (PrP(C)) expression can be regulated by heat-shock stress, and we designed the present study to determine whether hypoglycemia could affect PrP(C) expression. RT-PCR and Western blotting were used to measure the expression of PrP(C) and heat-shock protein (Hsp70) in mouse neuroblastoma (N18) cells cultured 3 hr to 3 days in media deprived of 97.5% (L) or 75% (M) of its glucose. Hypoglycemia caused a concomitant time-dependent and glucose dose dependent increase in PrP(C) and Hsp70. In addition, hypoglycemia also increased phosphorylated c-Jun N-terminal kinase (JNK) protein levels in a time-dependent manner. The upregulation of PrP(C) and Hsp70 under hypoglycemic conditions was disrupted by the specific JNK inhibitor SP600125. It was also found from in vitro studies that hypoglycemic conditions induced higher levels of PrP(C) promoter activity in PrP(C) promoters containing a heat-shock element (HSE) than in PrP(C) promoters lacking HSE. We propose that hypoglycemia-increased PrP(C) expression might be due to JNK phosphorylation of a heat-shock transcriptional factor, which then interacts with HSE in the promoter of PrP(C). PMID- 15884020 TI - Functional interactions between oculomotor regions during prosaccades and antisaccades. AB - Human behavior reflects a continual negotiation of automatic and directed actions. The oculomotor network is a well-characterized neural system in which to study this balance of behavioral control. For instance, saccades made toward and away from a flashed visual stimulus (prosaccades and antisaccades, respectively) are known to engage different cognitive processes. Brain regions important for such controlled execution include the presupplementary motor area (pre-SMA), frontal eye fields (FEF), and intraparietal sulcus (IPS). Recent work has emphasized various elements of this network but has not explored the functional interactions among regions. We used event-related fMRI to image human brain activity during performance of an interleaved pro/antisaccade task. Since traditional univariate statistics cannot address issues of functional connectivity, a multivariate technique is necessary. Coherence between fMRI time series of the pre-SMA with the FEF and IPS was used to measure functional interactions. The FEF, but not IPS, showed significant differential coherence between pro- and antisaccade trials with pre-SMA. These results suggest that the pre-SMA coordinates with FEF to maintain a controlled, preparatory set for task appropriate oculomotor execution. PMID- 15884021 TI - Mice with the deleted neurofilament of low molecular weight (Nefl) gene: 2. Effects on motor functions and spatial orientation. AB - Mice with a null mutation of the Nefl gene were compared with normal controls in tests of motor activity, equilibrium, and spatial orientation. Despite a normal capacity to ambulate, NFL -/- mice had fewer rears in an open field, crossed fewer segments on stationary beams, and fell more frequently when suspended on a horizontal bar. In addition, the distance swum before reaching the escape platform was greater in NFL -/- mice than in controls during acquisition of place learning in the Morris water maze at the start of training. The motor impairments were linearly correlated with increased cytochrome oxidase activity seen in cerebellum and brainstem. These results indicate that, as early as 6 months, depletion of the NFL protein is sufficient to cause mild sensorimotor dysfunctions and spatial deficits, but without overt signs of paresis. PMID- 15884022 TI - Prevalence and characteristics of nursing homes residents requiring light-care. AB - Rising nursing home (NH) costs and the poor quality of NH care make it important to recognize elders for whom NH care may be inappropriate. As a first step in developing a method to identify these elders, we examined the characteristics of NH residents requiring light-care and changes in their care level from NH admission to 12-months. Using data from the Missouri Minimal Data Set electronic database, we developed three care-level categories based on Resource Use Groups, Version III (RUG-III) and defined light-care NH residents as those requiring minimal assistance with late-loss ADLs (bed mobility, transfer, toilet use, or eating) and having no complex clinical problems. Approximately 16% of Missouri NH residents met the criteria for light-care. They had few functional problems with mobility, personal care, communication, or incontinence; approximately 33% had difficulty maintaining balance without assistance; and 50% of those admitted as light-care were still light-care at 12-months. Findings suggest that many NH residents classified as light-care by these criteria could be cared for in community settings offering fewer services than NHs. PMID- 15884023 TI - Neural correlates of the misattribution of self-generated speech. AB - Auditory hallucinations are thought to arise through the misidentification of self-generated verbal material as alien. The neural mechanisms that normally mediate the differentiation of self-generated from nonself speech are unclear. We investigated this in healthy volunteers using functional MRI. Eleven healthy volunteers were scanned whilst listening to a series of prerecorded words. The source (self/nonself) and acoustic quality (undistorted/distorted) of the speech was varied across trials. Participants indicated whether the words were spoken in their own or another person's voice via a button press. Listening to self generated words was associated with more activation in the left inferior frontal and right anterior cingulate cortex than words in another person's voice, which was associated with greater engagement of the lateral temporal cortex bilaterally. Listening to distorted speech was associated with activation in the inferior frontal and anterior cingulate cortex. There was an interaction between the effects of source of speech and distortion on activation in the left temporal cortex. In the presence of distortion participants were more likely to misidentify their voice as that of another. This misattribution of self-generated speech was associated with reduced engagement of the cingulate and prefrontal cortices. The evaluation of auditory speech involves a network including the inferior frontal, anterior cingulate, and lateral temporal cortex. The degree to which different areas within this network are engaged varies with the source and acoustic quality of the speech. Accurate identification of one's own speech appears to depend on cingulate and prefrontal activity. PMID- 15884024 TI - Effect of guided participation on feeding competencies of mothers and their premature infants. AB - The effect of guided participation (GP) on premature infant and maternal feeding competencies was examined, controlling for infant, maternal, and family conditions. Competencies were examined longitudinally and within age (1, 4, 8, and 12 months post-term age) for 42 mother-infant pairs randomly assigned to either GP or Standard Care (SC) groups. The hypothesized GP effect on competencies across infant age received support for infants (at 1 and 8 months) and for mothers (at 4 months). The hypothesized contribution of conditions was most strongly supported by the negative relationship between family poverty status and the two maternal competency variables. The hypothesized GP moderator effect on the relationship between depressive symptoms and maternal competency variables was supported for regulation of negative affect and behavior at 8 months. Despite study limitations in power and sensitivity to detect effects, findings indicate that further study of the GP intervention is merited. PMID- 15884025 TI - Qualitative research contribution to a randomized clinical trial. AB - Qualitative research may be combined fruitfully with intervention studies, but few examples provide detailed methodological strategies for doing so. In this article, we describe the qualitative component of a randomized clinical trial (RCT) of the PRO-SELF(c) Pain Control Program, an intervention that provides individualized education, coaching, and support for cancer pain management. We conducted three qualitative analyses of verbatim transcripts of "real-time" audiotaped intervention sessions. As a result, we were better able to ascertain the nature of the individualized coaching component of the intervention, patient and family caregiver use of selected intervention tools, and reasons the intervention did not work for some patients. Study results were used to increase the specificity with which the coaching portion of the intervention is described in the intervention protocol. PMID- 15884026 TI - Healthy aging demonstration project: nurse coaching for behavior change in older adults. AB - The Healthy Aging Project (HAP) tested nurse coaching as a method to support healthy behavior change in older adults. The sample included 111 individuals randomized to a nurse coaching group or usual-care control group. Participants in the intervention group chose the health behaviors they wanted to change and received coaching by nurses in a single in-person session followed by telephone calls or email contact for 6 months. Nurses were trained in motivational interviewing (MI). The intervention group had significantly less illness intrusiveness and health distress than the control group at 6 months, although it is not known whether these health outcomes resulted from behavior changes. This clinical demonstration project showed that nurse-delivered MI, primarily using the telephone and email, is a feasible method to discuss behavioral change with older adults. However, future clinical trials will be needed to evaluate the efficacy of nurse-delivered MI on actual behavioral changes in older adults. PMID- 15884027 TI - Relationship of age, injury severity, injury type, comorbid conditions, level of care, and survival among older motor vehicle trauma patients. AB - The purpose of this secondary data analysis was to compare age, injury severity, injury types, selected comorbidities, level of care (at trauma center [TC] and non-trauma center [NTC] hospitals), and survival among older motor vehicle trauma patients (N = 1,478). Patients admitted to both levels of care had similar comorbid conditions. TC patients had a higher injury severity, whereas NTC patients had a greater proportion of soft tissue injuries. Results of logistic regression analyses subsequent to group comparisons revealed that higher injury severity was associated with TC admission. The likelihood of TC admission of severely injured patients decreased in the presence of spinal, internal, and head injuries. Internal injuries, liver, renal, and cardiovascular diseases were associated with non-survival while hypertension was associated with survival. Special attention is needed when triaging older trauma patients because their injuries may be covert, thus putting them at risk for admission to a level of care that may be inappropriate given the extent of their injuries. PMID- 15884028 TI - Clusters of problem behaviors in adolescents. AB - This study was designed to examine clusters of problem behaviors in a sample from the National Longitudinal Study of Adolescent Health at two time points. The technique employed was a person-oriented approach, cluster analysis. Three clusters were identified, a normal behaviors cluster, a problem behaviors cluster, and a deviant behaviors cluster. The clusters were tested for stability and for their relationships to the demographic variables gender, race, age, and socioeconomic status. The mean values for most of the problem behaviors in the problem behaviors cluster were higher than for those in the normal behaviors cluster and lower than for those in the deviant behaviors cluster. Selling drugs and weapon use distinguished the deviant behaviors cluster from the other two. Different interventions probably will be required to address the needs of those in each of the different clusters. PMID- 15884029 TI - Relaxation and music reduce pain following intestinal surgery. AB - Three nonpharmacological nursing interventions, relaxation, chosen music, and their combination, were tested for pain relief following intestinal (INT) surgery in a randomized clinical trial. The 167 patients were randomly assigned to one of three intervention groups or control and were tested during ambulation and rest on postoperative days 1 and 2. Pain sensation and distress were measured with visual analog scales (VAS). Multivariate analysis of covariance showed significantly less post-test pain in the intervention groups than in the control group on both days after rest and at three of six ambulation post-tests (p = .024 .001), resulting in 16-40% less pain. Mixed effects after ambulation were due to the large variation in pain and difficulty relaxing while returning to bed; but post hoc explorations showed effects for those with high and low pain. These interventions are recommended along with analgesics for greater postoperative relief without additional side effects. PMID- 15884030 TI - I speak English, don't I? PMID- 15884031 TI - Predictors of screening results for depressive symptoms among homeless adults in Los Angeles with latent tuberculosis. AB - The purpose of this study was to examine predictors of screening results for depressive symptoms in a Los Angeles homeless population with latent tuberculosis (TB). Four hundred and fifteen homeless adults participating in a nurse case managed intervention were included in this analysis. Logistic regression results indicated that those who reported a physical health limitation, multiple sex partners, daily drug use, alcohol dependence, or not having completed high school, were more likely to screen positive. Social support from non-drug users was protective. Given the importance of adherence to TB treatment regimens, the high prevalence of a positive screening for depressive symptoms in the homeless and the potential for depression to reduce adherence rates, routine screening and treatment for depression in high risk homeless adults being treated for TB may be warranted. PMID- 15884032 TI - Characteristics of patients with chronic hepatitis C who develop hepatocellular carcinoma after a sustained response to interferon therapy. PMID- 15884034 TI - Late postnatal maturation of excitatory synaptic transmission permits adult-like expression of hippocampal-dependent behaviors. AB - Sensorimotor systems in altricial animals mature incrementally during early postnatal development, with complex cognitive abilities developing late. Of prominence are cognitive processes that depend on an intact hippocampus, such as contextual-configural learning, allocentric and idiocentric navigation, and certain forms of trace conditioning. The mechanisms that regulate the delayed maturation of the hippocampus are not well understood. However, there is support for the idea that these behaviors come "on line" with the final maturation of excitatory synaptic transmission. First, by providing a timeline for the first behavioral expression of various forms of learning and memory, this study illustrates the late maturation of hippocampal-dependent cognitive abilities. Then, functional development of the hippocampus is reviewed to establish the temporal relationship between maturation of excitatory synaptic transmission and the behavioral evidence of adult-like hippocampal processing. These data suggest that, in rats, mechanisms necessary for the expression of adult-like synaptic plasticity become available at around 2 postnatal weeks of age. However, presynaptic plasticity mechanisms, likely necessary for refinement of the hippocampal network, predominate and impede information processing until the third postnatal week. PMID- 15884035 TI - Retrieval of autobiographical memory in Alzheimer's disease: relation to volumes of medial temporal lobe and other structures. AB - The representation of autobiographical memory is distributed over a network of brain structures, with the medial temporal lobe (MTL) at its epicenter. Some believe that, over time, all memories become independent of their MTL component ("consolidation theories"). Others have suggested that this is true only of semantic memory, while episodic aspects of autobiographical memories are dependent on the MTL for as long as they exist, such as multiple trace theory (MTT). In the present study, the volumes of 28 brain regions, including the MTL, and their relation to autobiographical memory were investigated in a group of patients with Alzheimer's disease with varying degrees of retrograde memory loss as assessed by the Autobiographical Memory Interview (AMI). We used the multivariate analysis method of partial least squares (PLS) to assess patterns of atrophy that can lead to retrograde amnesia. We found that different aspects of autobiographical memory were associated with different patterns of tissue loss. Personal semantics were related to a pattern of bilateral anterior and posterior lateral temporal cortex degeneration, more pronounced on the left, as well as right frontal degeneration. Autobiographical event memory ("episodic") was associated with combined atrophy in bilateral MTL and anterior lateral temporal neocortex, more pronounced on the right. This pattern was invariant for memories from childhood, early adulthood, and recent memories, in line with the predictions of MTT, suggesting that MTL tissue is crucial for retrieval of episodic memories regardless of their age. PMID- 15884036 TI - Effects of periodic leg movements during sleep in middle-aged subjects without sleep complaints. AB - Recent reports have called into question the relevance of periodic leg movements during sleep disorder (PLMSD) as a specific clinical entity. Because periodic leg movement in sleep index (PLMSI) increases with age, it has become an important exclusion criterion in research on aging. However, it is unknown if PLMSI is related to sleep quality in middle-aged subjects without sleep complaints. The sleep of 70 healthy, middle-aged subjects (age 40 to 60 years) without sleep complaints was evaluated. Subjects were divided into two groups according to their PLMSI severity: (1) 43 subjects (28 women, 15 men) were in the low PLMSI group (<5) and (2) 22 subjects (9 women, 13 men) were in the high PLMSI group (>10). A significantly higher proportion of men than women showed PLMSI greater than 5. There was no significant effect of PLMSI severity group for polysomnographic sleep parameters. PLMSI exerted a small but significant effect on subjective sleep quality, especially in middle-aged men. These results raise questions about the relevance of PLMSI as a pathological index for middle-aged subjects without sleep complaints and support the notion that an increase in PLMSI may be part of the normal process of aging associated with the loss of dopaminergic function. PMID- 15884037 TI - Functional expression of the ecto-ATPase NTPDase2 and of nucleotide receptors by neuronal progenitor cells in the adult murine hippocampus. AB - An astrocyte-like cell population corresponding to residual radial glia represents the neuronal progenitors of the adult mammalian hippocampus. We show that radial glia-like cells of the dentate gyrus express surface-located ATP hydrolyzing activity and are immunopositive for NTPDase2. This enzyme hydrolyzes extracellular nucleoside triphosphates such as ATP and UTP to their nucleoside diphosphates and is thus involved in the control of signaling via P2 receptors. NTPDase2 is expressed from embryonic day 17 onward. In the hippocampus, the embryonic pattern of NTPDase2 expression mirrors that of the dentate migration of neuroblasts. Double-immunolabeling revealed that NTPDase2 is associated with subpopulations of glial fibrillary acidic protein-, nestin- and doublecortin positive radial cells. It is absent from mature granule cells or S100-positive astrocytes. NTPDase2-positive cells proliferate. Furthermore, after mitosis, progenitor cells preferentially reveal an NTPDase2-positive phenotype. Patch clamp analysis demonstrates functional nucleotide receptors in progenitor cells expressing nestin promotor-driven green fluorescent protein. Our results identify the ecto-nucleotidase NTPDase2 and functional P2X receptors at hippocampal progenitor cells. We infer that signaling pathways via extracellular nucleotides may play a role in the control of hippocampal neurogenesis. PMID- 15884038 TI - Emergency hospital admissions in idiopathic Parkinson's disease. AB - Little is known about the hospital inpatient care of patients with idiopathic Parkinson's disease (PD). Here, we describe the features of the emergency hospital admissions of a geographically defined population of PD patients over a 4-year period. Patients with PD were identified from a database for a Parkinson's disease service in a district general hospital with a drainage population of approximately 180,000. All admissions of this patient subgroup to local hospitals were found from the computer administration system. Two clinicians experienced in both general medicine and PD then reviewed the notes to identify reasons for admission. Admission sources and discharge destinations were recorded. Data regarding non-PD patients was compared to PD patients on the same elderly care ward over the same time period. The total number of patients exposed to analysis was 367. There was a total exposure of 775.8 years and a mean duration of 2.11 years per patient. There were 246 emergency admissions to the hospital with a total duration of stay of 4,257 days (mean, 17.3 days). These days were accounted for by 129 patients (mean age, 78 years; 48% male). PD was first diagnosed during 12 (4.9%) of the admissions. The most common reasons for admission were as follows: falls (n=44, 14%), pneumonia (n=37, 11%), urinary tract infection (n=28, 9%), reduced mobility (n=27, 8%), psychiatric (n=26, 8%), angina (n=21, 6%), heart failure (n=20, 6%), fracture (n=14, 4%), orthostatic hypotension (n=13, 4%), surgical (n=13, 4%), upper gastrointestinal bleed (n=10, 3%), stroke/transient ischemic attack (n=8, 2%), and myocardial infarction (n=7, 2%). The mean length of stay for the PD patients on the care of elderly ward specializing in PD care was 21.3 days compared to 17.8 days for non-PD patients. After hospital admission, there was a reduction in those who returned to their own home from 179 to 163 and there was an increase in those requiring nursing home care from 37 to 52. Infections, cardiovascular diseases, falls, reduced mobility, and psychiatric complications accounted for the majority of admissions. By better understanding the way people with PD use hospital services, we may improve quality of care and perhaps prevent some inpatient stays and care-home placements. PMID- 15884039 TI - Evaluation of acupuncture in the treatment of Parkinson's disease: a double-blind pilot study. AB - As many as 40% of patients with Parkinson's disease (PD) use some form of complementary medicine during the course of their illness, and many try acupuncture. One nonblinded study of the effects of acupuncture in PD suggested that it might be helpful for some aspects of PD. We performed a double-blind, randomized, pilot study comparing acupuncture to a control nonacupuncture procedure to determine the effects of acupuncture upon a variety of PD-associated symptoms. Fourteen patients with Stage II or III PD received acupuncture or a control nonacupuncture protocol. Before and after treatment, patients were evaluated using the Motor subscale of the Unified Parkinson's Disease Rating Scale (UPDRS), the Parkinson's Disease Questionnaire (PDQ-39), and the Geriatric Depression Scale. There were no statistically significant changes for the outcomes measured. In the patients who received acupuncture, nonsignificant trends toward improvement were noted in the Activities of Daily Living score of the PDQ-39, the PDQ-39 Summary Index(c) 2005 Movement Disorder Society. PMID- 15884040 TI - Sensitive multistep clinical molecular screening of 180 unrelated individuals with retinoblastoma detects 36 novel mutations in the RB1 gene. AB - Retinoblastoma (RB) is a neoplasm of retinal origin caused by mutations in RB1, the retinoblastoma tumor suppressor gene. To facilitate genetics counseling and patient management, we adopted a multistep molecular screening assay for detecting RB1 mutations. This assay included DNA sequencing to identify mutations within coding exons and immediate flanking intronic regions, Southern blot analysis to characterize genomic rearrangements, and transcript analysis to characterize potential splicing mutations buried within introns. In a pilot investigation of 180 patients from North America, we identified germline RB1 mutations in 77 out of 85 bilateral RB patients (91%), 7 out of 10 familial unilateral (70%), and 6 out of 85 unilateral patients with no family history of RB (7%). Mutations included 36 novel alterations spanning the entire RB1 gene. Seven of these novel changes were missense or silent mutations. Sequence analysis predicted that, in five out of seven cases, the changes can cause aberrant splicing. This was confirmed by transcript analysis in four out of five cases. In addition, four intronic point mutations within nonconsensus sites activated cryptic splice sites. Without the transcript analysis, the significance of these 11 mutations would have remained undefined. In a separate investigation of a subset of unilateral RB tumors, we identified somatic biallelic RB1 gene inactivation in 34 out of 56 cases (61%) cases. In 14 tumors, only one of the two RB1 mutations could be detected, and in eight tumors, no mutations were detected. The absence of detectable RB1 mutations in eight bilateral cases and eight unilateral tumors suggests that alternative genetic mechanisms may underlie the development of RB in certain individuals. PMID- 15884041 TI - Genetic testing in Parkinson's disease. PMID- 15884042 TI - Shprintzen-Goldberg syndrome: fourteen new patients and a clinical analysis. AB - The Shprintzen-Goldberg syndrome (SGS) is a disorder of unknown cause comprising craniosynostosis, a marfanoid habitus and skeletal, neurological, cardiovascular, and connective-tissue anomalies. There are no pathognomonic signs of SGS and diagnosis depends on recognition of a characteristic combination of anomalies. Here, we describe 14 persons with SGS and compare their clinical findings with those of 23 previously reported individuals, including two families with more than one affected individual. Our analysis suggests that there is a characteristic facial appearance, with more than two thirds of all individuals having hypertelorism, down-slanting palpebral fissures, a high-arched palate, micrognathia, and apparently low-set and posteriorly rotated ears. Other commonly reported manifestations include hypotonia in at least the neonatal period, developmental delay, and inguinal or umbilical hernia. The degree of reported intellectual impairment ranges from mild to severe. The most common skeletal manifestations in SGS were arachnodactyly, pectus deformity, camptodactyly, scoliosis, and joint hypermobility. None of the skeletal signs alone is specific for SGS. Our study includes 14 mainly German individuals with SGS evaluated over a period of 10 years. Given that only 23 other persons with SGS have been reported to date worldwide, we suggest that SGS may be more common than previously assumed. PMID- 15884043 TI - Hemispheric brain volume replacement with free latissimus dorsi flap as first step in skull reconstruction. AB - Large skull defects lead to progressive depression deformities, with resulting neurological deficits. Thus, cranioplasty with various materials is considered the first choice in therapy to restore cerebral function. A 31-year-old female presented with a massive left-sided hemispheric substance defect involving bone and brain tissue. Computed tomography showed a substantial convex defect involving the absence of calvarial bone as well as more than half of the left hemisphere of the brain, with a profound midline shift and a compression of the ventricular system. There was a severe problem due to multiple deep-skin ulcerations at the depression margin, prone to skin perforation with a probability of intracranial infection. In a first step, a free myocutaneous latissimus dorsi flap was transplanted for volume replacement of the hemispheric brain defect, and 4 months later, artificial bone substitute was implanted in order to prevent progressive vault depression deformity. Healing was uneventful, and the patient showed definite neurological improvement postoperatively. Free tissue transfer can be a valuable option in addition to cranioplasty in the treatment of large bony defects of the skull. Besides providing stable coverage for the reconstructed bone or its substitute, it can also serve as a volume replacement. PMID- 15884044 TI - Hippocampal nitric oxide synthase and arginase and age-associated behavioral deficits. AB - The present study investigated age-related changes in nitric oxide synthase (NOS) and arginase in the subregions of the hippocampus and their correlations with animals' performance in the open field, T-maze, and water maze tasks. Aged rats (24 months old) showed reduced exploratory activity and poorer spatial learning relative to the young adults (4 months old). Significant increases in total NOS activity were found in the aged dentate gyrus and a dramatic decrease in endothelial NOS expression was observed in the aged CA2/3. Activity or protein expression of inducible NOS was not detected in any subregion of the hippocampus. There were no age-related changes in total arginase activity or arginase I and arginase II protein expression. Correlation analysis revealed that animals' motor ability was associated with CA1 NOS and arginase, as well as hippocampal function. The present findings provide further support for the involvement of NOS/NO and arginase in the normal aging process. A strong positive correlation between CA1 eNOS protein expression and swimming speed in the water maze task may reflect a relationship between the local cerebral blood flow and neuronal activity. PMID- 15884045 TI - Association of migraine-like headaches with Schimke immuno-osseous dysplasia. AB - Schimke immuno-osseous dysplasia (SIOD) is characterized by spondyloepiphyseal dysplasia, nephropathy, and T-cell deficiency. SIOD is caused by mutations in the putative chromatin remodeling protein SMARCAL1. We report an 8-year-old boy with SIOD and recurrent, severe, refractory migraine-like headaches. Through a retrospective questionnaire-based study, we found that refractory and severely disabling migraine-like headaches occur in nearly half of SIOD patients. We have also found that the vasodilator minoxidil provided symptomatic relief for one patient. We hypothesize that these headaches may arise from an intrinsic vascular, neuroimmune, or neurovascular defect resulting from loss of SMARCAL1 function. PMID- 15884047 TI - Hippocampal theta activity and behavioral sequences in a reward-directed approach locomotor task. AB - Hippocampal rhythmic slow wave activity (theta) has been implicated in the processing of stimuli associated with movement. This study determined whether the theta rhythm showed phase relationships or changes in amplitude and frequency with the onset of stimuli and behavioral sequences in a skilled locomotor approach task. Rats with bipolar electrodes spanning CA1 approached a stall, turned to enter it, approached and depressed a treadle, waited 1.35 s, and approached a milk reward located forward either to the right or to the left. Auditory cues indicated the location of the reward during the waiting period and at the reward onset. A video capture system (20-ms sampling) was synchronized to the hippocampal recording system (10-ms sampling). Behavioral events identified by motion analysis were used to generate averages of hippocampal slow wave activity, theta peak amplitudes, and intervals between peaks. Theta activity at 8 10 Hz was almost continuous during the behavioral sequences. Phase relations with stimuli or movement onsets occurred infrequently and were not consistent across the four subjects. Theta peak amplitude and frequency decreased as the rat slowed locomotion in the stall and reached the treadle. Onset of locomotion directed to a reward location occurred on a positive peak of averaged theta activity. When locomotion had short latencies, increases in theta frequency appeared after the onset but, when it had longer latencies, frequency increases appeared 200 ms before onset. The results indicate that the execution of instrumental movement modulates both theta amplitude and frequency, and that the preparation for locomotion modulates theta frequency. PMID- 15884048 TI - Ubiquitin C-terminal hydrolase L3 (Uchl3) is involved in working memory. AB - Regulated proteolysis by the ubiquitin pathway has been implicated in control of the cell cycle, transcriptional activation, cell fate and growth, and synaptogenesis. The ubiquitin-proteasome system is involved in synaptic plasticity and is proposed to be part of a molecular switch that converts short term synaptic potentiation to long-term changes in synaptic strength. In Aplysia, a component of the ubiquitin system termed ubiquitin C-terminal hydrolase (Ap Uch) has been shown to be essential for long-term facilitation. To examine whether Uch plays a role in learning, memory, and synaptic plasticity in mammals, we have analyzed mice homozygous for a targeted mutation in ubiquitin C-terminal hydrolase L3 (Uchl3), an orthologue of Ap-Uch. Mice homozygous for the mutation in Uchl3 are viable, with no obvious developmental, histological, or fertility abnormalities. We demonstrate that Uchl3-/- mice have a significant learning deficit relative to wild type littermates in the spatial version of the Morris water maze and the 8-arm radial maze. Further, the impaired performance in the 8 arm radial maze of Uchl3-/- mice is due to significantly increased working memory errors. Examination of hippocampal long-term potentiation (LTP), a form of synaptic plasticity thought to underlie memory storage, revealed no significant differences in LTP in hippocampal slices from Uchl3-/- mice. Our results suggest a novel role for ubiquitin C-terminal hydrolase enzymes in mammals in spatial learning and working memory. PMID- 15884050 TI - Human CD8+ T cell blasts are more sensitive than CD4+ T cell blasts to regulation by APO2L/TRAIL. AB - The mechanisms responsible for the down-modulation of the activation of separated CD4(+) or CD8(+) human T cell blasts were studied using cells obtained from healthy donors. In the presence of IL-2, human CD8(+) T cell blasts were more sensitive than CD4(+) T cell blasts to regulation by APO2 ligand/TNF-related apoptosis-inducing ligand (APO2L/TRAIL), while both T cell subsets were equally sensitive to Fas/CD95 regulation. This regulation was defined as inhibition of IL 2-dependent T cell growth in the absence of cell death induction, characterized by cell cycle arrest in G(2)/M. The physiological validity of these observations was corroborated by the demonstration of intracellular FasL and APO2L/TRAIL expression in CD4(+) and CD8(+) T cell blasts, which were secreted in their bioactive form into the supernatant upon PHA, CD3 or CD59 reactivation. Additionally, the inhibition of IL-2-dependent CD4(+) or CD8(+) T cell blast growth upon CD3 or CD59 ligation was dependent, at least partially, on FasL and/or APO2L/TRAIL. These data precisely define the role of APO2L/TRAIL in the regulation of human T cell activation. PMID- 15884052 TI - beta1 integrins are not required for the maintenance of lymphocytes within intestinal epithelia. AB - beta(1) integrins are thought to play a central role in maintaining lymphocytes within mucosal epithelia via their interactions with extracellular matrix proteins and subepithelial cellular components within and underlying the basement membrane. In the current study type a (CD8alphabetaTCRalphabeta) and type b (CD8alphaalphaTCRgammadelta and CD8alphaalphaTCRalphabeta) intraepithelial lymphocyte (IEL) subsets within the mouse small intestine were found to express functional beta(1) integrin and the beta(1) integrin alpha chain partners alpha(1), alpha(2), and alpha(4). Using inducible beta(1) integrin-knockout bone marrow-chimeric mice we demonstrate that IEL expression of alpha(1) and alpha(2) but not alpha(4) is dependent on expression of the beta(1) chain. Importantly, deletion of the beta(1) chain in IEL did not alter the number or composition of lymphocytes within the intestinal epithelium. Thus, while IEL express functional beta(1) integrins, these are not required to maintain lymphocytes within intestinal epithelia. This result is discussed in the light of conventional views of intestinal lymphocyte homing and localization. PMID- 15884053 TI - Antagonism between MyD88- and TRIF-dependent signals in B7RP-1 up-regulation. AB - Type I interferons (IFN) play a critical role in the Toll-like receptor (TLR) mediated expression of B7 costimulatory family members. For example, LPS-induced up-regulation of CD80 (B7.1) and CD86 (B7.2) is abrogated in antigen-presenting cells (APC) deficient in TRIF or TRAM, two adaptors that are responsible for TLR4 mediated production of Type I IFN. In this report, we demonstrate that LPS induced up-regulation of B7-related protein 1 (B7RP-1), a ligand for ICOS, is dependent primarily upon the MyD88-dependent signaling pathway. Signaling via the TRIF pathway sharply limits MyD88-dependent B7RP-1 up-regulation. Hence, LPS induces significantly higher B7RP-1 expression on TRIF- or TRAM-deficient mouse peritoneal macrophages and on TRIF-deficient mouse splenic B cells as compared to wild-type cells. Further studies reveal that Type I IFN are general suppressors of TLR-mediated up-regulation of B7RP-1. These data indicate that Type I IFN play a dual role in the TLR-mediated expression of B7 costimulatory family members and suggest that they may act to limit B7RP-1 expression and thus limit signals derived from B7RP-1-ICOS interaction. PMID- 15884054 TI - CXCR3 is required for migration to dermal inflammation by normal and in vivo activated T cells: differential requirements by CD4 and CD8 memory subsets. AB - Lymphocytes in inflamed tissues express numerous chemokine receptors. The relative importance of these receptors for migration in inflammation is unclear. The role of CXCR3 in T cell subset migration was examined using monoclonal antibodies developed to rat CXCR3. CXCR3 was expressed on sixfold more CD8(+) ( approximately 30%) than CD4(+) ( approximately 5%) T cells in spleen, lymph nodes and blood, and on approximately 10% of CD4(+)CD45RC(-) (memory) and approximately 50% of CD8(+)CD45RC(+) spleen T cells. After immunization, CXCR3 increased tenfold on CD4(+) lymph node lymphoblasts ( approximately 55%), and >90% of inflammatory exudate T cells were CXCR3(+). CXCR3(+) T cells migrated significantly better than CXCR3(-) T cells to all dermal inflammatory stimuli tested in vivo, even though these T cells are a minority of the memory T cells. Blocking CXCR3 inhibited recruitment of 60-85% of unstimulated T cells and up to 90% of CD8(+)CD45RC(+) effector T cells, but caused <50% inhibition of CD4(+) and CD8(+) memory (CD45RC(-)) T cells. About 90% of T lymphoblast migration to IFN gamma, IFN-gamma plus TNF-alpha, polyinosinic polycytidylic acid, lipopolysaccharide, and delayed-type hypersensitivity (DTH)-induced inflammation was inhibited. Blockade also reduced DTH-induced induration. Thus, CXCR3 has a non-redundant role in T cell migration to dermal inflammation and is critical for activated T lymphoblast recruitment, but memory T cells are less dependent on CXCR3 for their infiltration. PMID- 15884055 TI - KARAP/DAP12/TYROBP: three names and a multiplicity of biological functions. AB - The signaling adaptor protein KARAP/DAP12/TYROBP (killer cell activating receptor associated protein / DNAX activating protein of 12 kDa / tyrosine kinase binding protein) belongs to the family of transmembrane polypeptides bearing an intracytoplasmic immunoreceptor tyrosine-based activation motif (ITAM). This adaptor, initially characterized in NK cells, is associated with multiple cell surface activating receptors expressed in both lymphoid and myeloid lineages. We review here the main features of KARAP/DAP12, describing findings from its identification to recently published data, showing its involvement in a broad array of biological functions. KARAP/DAP12 is a wiring component for NK cell anti viral function (e.g. mouse cytomegalovirus via its association with mouse Ly49H) and NK cell anti-tumoral function (e.g. via its association with mouse NKG2D or human NKp44). KARAP/DAP12 is also involved in inflammatory reactions via its coupling to myeloid receptors, such as the triggering receptors expressed by myeloid cells (TREM) displayed by neutrophils, monocytes/macrophages and dendritic cells. Finally, bone remodeling and brain function are also dependent upon the integrity of KARAP/DAP12 signals, as shown by the analysis of KARAP/DAP12-deficient mice and KARAP/DAP12-deficient Nasu-Hakola patients. PMID- 15884056 TI - IFN-gamma production by innate immune cells is sufficient for development of hypersensitivity pneumonitis. AB - Hypersensitivity pneumonitis (HP) is an interstitial lung disease that develops following repeated exposure to inhaled particulate antigens. The disease is characterized by lymphocytic alveolitis, granuloma formation and fibrosis. IFN gamma is required for the formation of granulomas in HP, and we therefore focused on identifying the cellular sources of IFN-gamma during the disease. Using the Saccharopolyspora rectivirgula (SR) animal model of HP, we demonstrated that the majority of IFN-gamma(+) cells in the lung following SR exposure are neutrophils. Ab-mediated depletion of neutrophils in mice prior to exposure to SR resulted in a decrease in the level of IFN-gamma mRNA and protein compared to isotype Ab treated mice, suggesting that neutrophils are an important source of IFN-gamma during HP. To determine the contribution of T and non-T cell sources of IFN-gamma to granuloma formation, we performed adoptive transfer studies. RAG-1(-/-) mice reconstituted with spleen cells from IFN-gamma(-/-) mice developed granulomas similarly to RAG-1(-/-) mice reconstituted with normal spleen cells. Therefore innate immune cell IFN-gamma production in the absence of T cell IFN-gamma production is sufficient for granuloma formation. These results provide new insight into the pathogenesis of HP and demonstrate the important contribution of innate immune cells to the disease process. PMID- 15884057 TI - The Lsc RhoGEF mediates signaling from thromboxane A2 to actin polymerization and apoptosis in thymocytes. AB - The Lsc RhoGEF (also known as p115-RhoGEF) is a GTP exchange factor (GEF), an activator of GTPases of the Rho family. Lsc has a RhoGEF domain specific for Rho GTPase and a regulator of G protein signaling (RGS) domain specific for Galpha(12/13) subunits. One G protein receptor that can couple to Galpha(12/13) subunits is the receptor for thromboxane A(2 )(TXA(2)), thromboxane-prostanoid (called TP), which is highly expressed in immature thymocytes. TXA(2) has been implicated in thymocyte apoptosis. We found that Lsc(-/-) mice on a BALB/c background show thymic hyperplasia due to increased numbers of thymocytes and that these numbers further increase with the age of the mice. To investigate a role for Lsc in TXA(2) signaling, we analyzed activation of primary thymocytes by TXA(2) in vitro. TXA(2)-induced apoptosis of double-positive thymocytes and Rho activation required Lsc, and TXA(2) stimulation of actin polymerization and cofilin phosphorylation required both Lsc and Rho kinase (ROCK). Additionally, in the absence of Lsc, phosphorylation of the survival kinase Akt in response to TXA(2) was greatly enhanced. Together, these data demonstrate that Lsc is essential for mediating TXA(2 )signaling involved in apoptosis and actin organization and suggest that TXA(2) regulates thymic cellularity via Lsc. PMID- 15884058 TI - The coming of age. PMID- 15884060 TI - Statistical evaluation of single nano-object fluorescence. AB - Single nano-objects display strong fluctuations of their fluorescence signals. These random and irreproducible variations must be subject to statistical analysis to provide microscopic information. We review the main evaluation methods used so far by experimentalists in the field of single-molecule spectroscopy: time traces, correlation functions, distributions of "on" and "off" times, higher-order correlations. We compare their advantages and weaknesses from a theoretical point of view, illustrating our main conclusions with simple numerical simulations. We then review experiments on different types of single nano-objects, the phenomena which are observed and the statistical analyses applied to them. PMID- 15884061 TI - Molecular photobleaching kinetics of Rhodamine 6G by one- and two-photon induced confocal fluorescence microscopy. AB - Under high-excitation irradiance conditions in one- and two-photon induced fluorescence microscopy, the photostability of fluorescent dyes is of crucial importance for the detection sensitivity of single molecules and for the contrast in fluorescence imaging. Herein, we report on the dependence of photobleaching on the excitation conditions, using the dye Rhodamine 6G as a typical example. The different excitation modes investigated include 1) one-photon excitation into the first-excited singlet state in the range of 500 to 528 nm by continuous wave and picosecond-pulsed lasers and 2) two- and one-photon excitation to higher-excited singlet states at 800 and 350 nm, respectively, by femtosecond pulses. Experimental strategies are presented, which allow resolving the photophysics. From single-molecule trajectories and fluorescence correlation spectroscopy, as well as with a simple theoretical model based on steady-state solutions of molecular rate equation analysis, we determined the underlying photobleaching mechanisms and quantified the photokinetic parameters describing the dependence of the fluorescence signal on the excitation irradiance. The comparison with experimental data and an exact theoretical model show that only minor deviations between the different theoretical approaches can be observed for high-pulsed excitation irradiances. It is shown that fluorescence excitation is in all cases limited by photolysis from higher-excited electronic states. In contrast to picosecond-pulsed excitation, this is extremely severe for both one- and two photon excitation with femtosecond pulses. Furthermore, the photostability of the higher-excited electronic states is strongly influenced by environmental conditions, such as polarity and temperature. PMID- 15884062 TI - Intramolecular electron transfer in nitrite reductases. AB - The copper- and heme-containing nitrite reductases (NiRs) are key enzymes in denitrification. Their subunits contain two distinct redox-active metal centers, an electron-accepting site and a nitrite-reducing site, to carry out the single electron reduction of nitrite to nitric oxide. Catalytic cycles of both enzyme families employ intramolecular electron transfer that can be rate-determining for their activity. Herein, we report results comparing these two enzyme families in order to resolve the different mechanisms controlling intramolecular electron transfer in these proteins. PMID- 15884063 TI - Single-molecule manipulation measurements of DNA transport proteins. AB - Single-molecule measurements of the manipulation of three different DNA motor proteins are reviewed. Despite some differences in the structure and mechanisms of the proteins, there are consistent phenomenological themes that relate them. Each of the experiments described represents a significant advance in the understanding of the mechanisms of DNA transport. PMID- 15884064 TI - Energy transfer in single-molecule photonic wires. AB - Molecular photonics is a new emerging field of research around the premise that it is possible to develop optical devices using single molecules as building blocks. Truly technological impact in the field requires focussed efforts on designing functional molecular devices as well as having access to their photonic properties on an individual basis. In this Minireview we discuss our approach towards the design and single-molecule investigation of one-dimensional multimolecular arrays intended to work as molecular photonic wires. Three different schemes have been explored: a) perylene-based dimer and trimer arrays displaying coherent exciton delocalisation at room temperature; b) DNA-based unidirectional molecular wires containing up to five different chromophores and exhibiting weak excitonic interactions between neighbouring dyes; and c) one dimensional multichromophoric polymers based on perylene polyisocyanides showing excimerlike emission. As a whole, our single-molecule data show the importance of well-defined close packing of chromophores for obtaining optimal excitonic behaviour at room temperature. Further improvement on (bio)chemical synthesis, together with the use of single-molecule techniques, should lead in the near future to efficient and reliable photonic wires with true device functionality. PMID- 15884065 TI - Initial photoinduced dynamics of the photoactive yellow protein. AB - The photoactive yellow protein (PYP) is the photoreceptor protein responsible for initiating the blue-light repellent response of the Halorhodospira halophila bacterium. Optical excitation of the intrinsic chromophore in PYP, p-coumaric acid, leads to the initiation of a photocycle that comprises several distinct intermediates. The dynamical processes responsible for the initiation of the PYP photocycle have been explored with several time-resolved techniques, which include ultrafast electronic and vibrational spectroscopies. Ultrafast electronic spectroscopies, such as pump-visible probe, pump-dump-visible probe, and fluorescence upconversion, are useful in identifying the timescales and connectivity of the transient intermediates, while ultrafast vibrational spectroscopies link these intermediates to dynamic structures. Herein, we present the use of these techniques for exploring the initial dynamics of PYP, and show how these techniques provide the basis for understanding the complex relationship between protein and chromophore, which ultimately results in biological function. PMID- 15884066 TI - Green and red fluorescent proteins: photo- and thermally induced dynamics probed by site-selective spectroscopy and hole burning. AB - The cloning and expression of autofluorescent proteins in living matter, combined with modern imaging techniques, have thoroughly changed the world of bioscience. In particular, such proteins are widely used as genetically encoded labels to track the movement of proteins as reporters of cellular signals and to study protein-protein interactions by fluorescence resonance energy transfer (FRET). Their optical properties, however, are complex and it is important to understand these for the correct interpretation of imaging data and for the design of new fluorescent mutants. In this Minireview we start with a short survey of the field and then focus on the photo- and thermally induced dynamics of green and red fluorescent proteins. In particular, we show how fluorescence line narrowing and high-resolution spectral hole burning at low temperatures can be used to unravel the photophysics and photochemistry and shed light on the intricate electronic structure of these proteins. PMID- 15884067 TI - Coherent control for spectroscopy and manipulation of biological dynamics. AB - Motivated originally by the goal of steering a photoreaction into desired product channels, the concept of coherent control is to adapt the spectral and temporal characteristics of the excitation light to the inherent molecular resonances and dynamics, such that these can be selectively addressed and manipulated. In the last decade, the ultrafast dynamics of many atomic and molecular quantum systems in the gas and condensed phase have been controlled successfully. Motivations in chemistry are now 1) to perform spectroscopy by coherent control, which requires a deeper understanding of control mechanisms, 2) to treat more complex, biological photoreactions, and 3) the pragmatic use of coherent control techniques, for example, for pulse compression or enhanced contrast in multiphoton microscopy. As examples for 1) and 2) we review here the combined effort and interplay of conventional spectroscopy and coherent control experiments, applied to the energy flow in the light-harvesting complex LH2 from bacterial photosynthesis. Closed-loop control experiments allowed the characteristic coupling frequency of internal conversion in the carotenoid in LH2 to be extracted. Open-loop three-pulse control experiments, on the other hand, could directly observe an anticipated Raman-excited carotenoid ground state. As a variant of difference spectroscopy, coherent control has thus served to gain complementary spectroscopic knowledge about the energy flow in carotenoids by comparing natural to manipulated dynamics. Finally, we propose future coherent control experiments on the electronic state structure of carotenoids and discuss prospects of coherent control for other biological chromophores. PMID- 15884068 TI - The study of protein folding and dynamics by determination of intramolecular distance distributions and their fluctuations using ensemble and single-molecule FRET measurements. AB - The folding and dynamics of globular proteins is a multidimensional problem. The structures of the heterogeneous population of refolding protein molecules are characterized by multiple distances and time constants. Deciphering the mechanism of folding depends on studies of the processes rather than the folded structures alone. Spectroscopy is indispensable for these sorts of studies. Herein, it is shown that the determination of intramolecular distance distributions by ensemble and single-molecule FRET experiments enable the exploration of partially folded states of refolding protein molecules. PMID- 15884069 TI - The first picoseconds in bacterial photosynthesis--ultrafast electron transfer for the efficient conversion of light energy. AB - In this Minireview, we describe the function of the bacterial reaction centre (RC) as the central photosynthetic energy-conversion unit by ultrafast spectroscopy combined with structural analysis, site-directed mutagenesis, pigment exchange and theoretical modelling. We show that primary energy conversion is a stepwise process in which an electron is transferred via neighbouring chromophores of the RC. A well-defined chromophore arrangement in a rigid protein matrix, combined with optimised energetics of the different electron carriers, allows a highly efficient charge-separation process. The individual molecular reactions at room temperature are well described by conventional electron-transfer theory. PMID- 15884070 TI - Proteins in action monitored by time-resolved FTIR spectroscopy. AB - In the post genome era proteins coming into the focus of life sciences. X-ray structure analysis and NMR spectroscopy are established methods to determine the geometry of proteins. In order to determine the molecular reaction mechanism of proteins, time-resolved FTIR (trFTIR) difference spectroscopy emerges as a valuable tool. In this Minireview we describe the trFTIR difference spectroscopy and show its application on the light-driven proton pump bacteriorhodopsin (bR), the photosynthetic reaction center and the GTPase Ras, which is crucial in signal transduction. The main principles of the technique are presented, including a summary of triggering techniques, scan modes and analysis. PMID- 15884071 TI - Carriers versus adapters in stochastic sensing. PMID- 15884072 TI - Two-photon absorption of bis[4-(N,N-diphenylamino)phenylethynyl]arenes. PMID- 15884073 TI - Localization of single avidin-biotin interactions using simultaneous topography and molecular recognition imaging. PMID- 15884074 TI - Nanopatterning of biomolecules with microscale beads. PMID- 15884075 TI - Effect of bin time on the photon counting histogram for one-photon excitation. AB - We have demonstrated that our photon counting histogram (PCH) model with the correction for one-photon excitation is valid at multiple bin times. The fitted apparent brightness and concentration follow the three-dimensional diffusion model. More importantly, the semi-empirical parameter, F, introduced in the PCH model for one-photon excitation to correct for the non-Gaussian shape of the observation volume, shows small variations with different bin times. These variations are consistent with the physical interpretation of F, and they do not affect the resolving power of the PCH model for one-photon excitation. Based on these findings, we extend the time-independent PCH analysis to time-dependent photon counting multiple histograms (PCMH). This model considers the effect of bin time on the PCH parameters in a way that is similar to fluorescence intensity multiple distribution analysis (FIMDA). From the same set of data, PCMH extracts time-dependent parameters (diffusion time and triplet-state relaxation time) as well as time-independent parameters (true specific brightness and true average number of molecules). Given a three- to fourfold experimental difference in molecular brightness, we find that PCMH can resolve each species in a two-species sample and extract their respective diffusion times even when fluorescence correlation spectroscopy cannot. PMID- 15884076 TI - Surface-enhanced Raman scattering of p-aminothiophenol on a Au(core)/Cu(shell) nanoparticle assembly. AB - Surface-enhanced Raman scattering (SERS) of p-aminothiophenol (PATP) molecules adsorbed onto assemblies of Au(core)/Cu(shell) nanoparticles is reported. We compare it with the SERS spectrum of PATP adsorbed onto gold nanoparticles: both the absolute and relative scattered intensities of various bands in the two spectra are very different. The difference in relative intensity can be ascribed to chemical effects; the chemical enhancement ratio of the two substrates is approximately 3-5. A theoretical analysis based on a charge-transfer model is carried out, which yields a consistent result and shows that the difference in chemical enhancement is mainly due to the state densities and Fermi levels of the substrates. The difference in absolute intensity originates from electromagnetic (EM) enhancement. EM enhancement of Au(core)/Cu(shell) nanoparticles is unlike that of single-component gold or copper SERS-active substrates. The core/shell particle size for optimal enhancement is about 20 nm in the case of a 632.8 nm incident laser (the size ratio of the core and shell layers is about 0.6). PMID- 15884077 TI - Vibronic excitation of single molecules: a new technique for studying low temperature dynamics. AB - Herein, we present vibronic excitation and detection of purely electronic zero phonon lines (ZPL) of single molecules as a new tool for investigating dynamics at cryogenic temperatures. Applications of this technique to study crystalline and amorphous matrix materials are presented. In the crystalline environment, spectrally stable ZPLs are observed at moderate excitation powers. By contrast, investigations at higher excitation intensities reveal the opening of local degrees of freedom and spectral jumps, which we interpret as the observation of elementary steps in the melting of a crystal. We compare these results to spectral single-molecule trajectories recorded in a polymer. The way in which much more complicated spectral features can be analysed is shown. Surprisingly, pronounced spectral shifts on a previously not accessible large energy scale are observed, which are hard to reconcile with the standard two-level model system used to describe low-temperature dynamics in disordered systems. PMID- 15884078 TI - Single chromophore spectroscopy of MEH-PPV: homing-in on the elementary emissive species in conjugated polymers. AB - Low-temperature, single-molecule spectroscopy can provide unparalleled access to the primary emissive species of conjugated polymers. We demonstrate this with the example of one of the most commonly studied polymers, poly(2-methoxy-5-(2' ethylhexoxy)-1,4-phenylenevinylene), MEH-PPV, which is shown to exhibit sharp fluorescence signatures over one hundred times narrower than the ensemble. These unprecedented narrow emission features can be assigned to single chromophores on the polymer chain, which are selectively addressed by the narrow band excitation. As with organic dye systems, the emission from single chromophores is not static with time, but shows a substantial spectral fluctuation. We find that, for single chromophores, this spectral fluctuation always follows a universal Gaussian statistical distribution. High-resolution spectroscopy provides unique insight into low-energy vibrational modes in the polymer emission, which are generally inaccessible with conventional spectroscopic methods such as site-selective fluorescence or Raman spectroscopy. Interchromophoric coupling can also occur owing to the flexible nature of the polymer backbone. This leads to substantial spectral broadening and a loss of resolution in the vibronic progression. We observe reversible switching within one single molecule between narrow and broad emission, which directly correlates with a discrete switching in emission intensity. We conclude that one and the same single molecule can support aggregated and nonaggregated emission, that is, emission from isolated and aggregated chromophores in one single molecule, rather than the tendency for aggregate emission being intrinsic to the molecule. PMID- 15884079 TI - Photophysical properties of a tetraphenoxy-substituted perylene bisimide derivative characterized by single-molecule spectroscopy. AB - We present a detailed study of the photophysical properties of a tetraphenoxy substituted perylene bisimide derivative. The probe molecules were immobilized in a Shpol'skii matrix of hexadecane and investigated by single-molecule spectroscopy at cryogenic temperatures. By using single-molecule spectroscopic techniques we reveal the triplet substate kinetics and the fluorescence quantum yield, and we provide an estimate for the S1-S0 transition dipole moment. PMID- 15884080 TI - Electron transfer at the single-molecule level in a triphenylamine-perylene imide molecule. AB - Photoinduced electron transfer (ET) processes in a donor-acceptor system based on triphenylamine and perylene imide have been studied at the single-molecule (SM) and ensemble levels. The system exists as two isomers, one of which undergoes forward and reverse ET in toluene with decay constants of 3.0 and 2.2x10(9) s( 1), respectively, resulting in the dual emission of quenched and delayed fluorescence while the other isomer remains ET-inactive. The fluorescence of both isomers is heavily quenched in the more polar solvent, diethyl ether, by ET. A broad range of ET dynamics is seen at the SM level in polystryene with the two isomers nonresolvable indicating that the local nanoenvironment of the SMs varies considerably throughout the polymer matrix. Both the electronic coupling and the driving force for ET are shown to influence the ET dynamics. Many fluorescence trajectories of SMs show long periods (tens of milliseconds to seconds) where the count rate is attenuated either partly (a "dim" state) or to the background level (an "off-time"). During these periods, the reduction or interruption of emission is attributed to cycles of rapid charge separation followed by charge recombination to the ground state reducing the fluorescence quantum yield of the SM. PMID- 15884081 TI - High-resolution colocalization of single molecules within the resolution gap of far-field microscopy. AB - To obtain detailed information about the three-dimensional (3D) organization of small biomolecular assemblies with a size of less than 100 nanometers, advanced techniques are required that enable the determination of absolute 3D positions and distances between individual fluorophores well below the resolution limit of conventional light microscopy. We show how spectrally resolved fluorescence lifetime imaging microscopy (SFLIM) can provide significant contributions and allow us to determine distances between conventional individual fluorophores (Bodipy 630/650 and Cy5.5) that are less than 20 nm apart. We take advantage of fluorescent dyes (here Cy5.5 and Bodipy 630/650) that can be efficiently excited by a single pulsed diode laser emitting at 635 nm but differ in their fluorescence lifetime and emission maxima. The potential of the method for ultrahigh colocalization studies is demonstrated by measuring the end-to-end distance between single fluorophores separated by double-stranded DNA of various lengths. Combining SFLIM with polarization-modulated excitation allows us to obtain, simultaneously, information about the relative orientation of fluorophores. Furthermore, we show that the environment-dependent photophysics of conventional fluorophores, that is, photostability, blinking pattern, and the tendency to enter irreversible nonfluorescent states, sets certain limitations to their in vitro and in vivo applications. PMID- 15884082 TI - Single-molecule quantum-dot fluorescence resonance energy transfer. AB - Colloidal semiconductor quantum dots are promising for single-molecule biological imaging due to their outstanding brightness and photostability. As a proof of concept for single-molecule fluorescence resonance energy transfer (FRET) applications, we measured FRET between a single quantum dot and a single organic fluorophore Cy5. DNA Holliday junction dynamics measured with the quantum dot/Cy5 pair are identical to those obtained with the conventional Cy3/Cy5 pair, that is, conformational changes of individual molecules can be observed by using the quantum dot as the donor. PMID- 15884083 TI - Electrochemical and spectroscopic investigations of immobilized de novo designed heme proteins on metal electrodes. AB - On the basis of rational design principles, template-assisted four-helix-bundle proteins that include two histidines for coordinative binding of a heme were synthesized. Spectroscopic and thermodynamic characterization of the proteins in solution reveals the expected bis-histidine coordinated heme configuration. The proteins possess different binding domains on the top surfaces of the bundles to allow for electrostatic, covalent, and hydrophobic binding to metal electrodes. Electrostatic immobilization was achieved for proteins with lysine-rich binding domains (MOP-P) that adsorb to electrodes covered by self-assembled monolayers of mercaptopropionic acid, whereas cysteamine-based monolayers were employed for covalent attachment of proteins with cysteine residues in the binding domain (MOP C). Immobilized proteins were studied by surface-enhanced resonance Raman (SERR) spectroscopy and electrochemical methods. For all proteins, immobilization causes a decrease in protein stability and a loosening of the helix packing, as reflected by a partial dissociation of a histidine ligand in the ferrous state and very low redox potentials. For the covalently attached MOP-C, the overall interfacial redox process involves the coupling of electron transfer and heme ligand dissociation, which was analyzed by time-resolved SERR spectroscopy. Electron transfer was found to be significantly slower for the mono-histidine coordinated than for the bis-histidine-coordinated heme. For the latter, the formal heterogeneous electron-transfer rate constant of 13 s(-1) is similar to those reported for natural heme proteins with comparable electron-transfer distances, which indicates that covalently bound synthetic heme proteins provide efficient electronic communication with a metal electrode as a prerequisite for potential biotechnological applications. PMID- 15884084 TI - Vibronic transitions in single metalloporphyrins. AB - Structural and electronic properties of single zinc etioporphyrin molecules adsorbed on Al2O3/NiAl(110) were probed by a low-temperature scanning tunneling microscope (STM). Scanning tunneling spectroscopy (STS) revealed progressions of spectral features corresponding to the vibronic states of individual molecules that depend strongly on the molecular conformations. Vibronic features observed by STS were compared with the results from fluorescence induced by tunneling electrons (tunneling-induced fluorescence, TIF). PMID- 15884085 TI - Optical characteristics of atomic force microscopy tips for single-molecule fluorescence applications. AB - Knowledge of the optical properties of atomic force microscopy (AFM) tips is relevant for the combination of optical and force spectroscopy. The luminescence properties of five commercial AFM tips were characterized using a combination of multiparameter fluorescence detection (MFD) and scanning confocal techniques. These include three Si3N4 tips, one silicon tip, and one high-density carbon (HDC) tip grown on top of a silicon tip. Time-decay histograms of the signal were analyzed to determine the strength of scatter, constant background, and fluorescence in the observed signal. Intensity and anisotropy images with optical resolution down to the diffraction limit were generated. The optical signal recorded from the apex of the Si3N4 tips ranged from 0.7 to 1.9 times the count rates from single Rhodamine 110 molecules under similar illumination conditions. The signal is predominantly composed of scatter and background (>85%), plus a small fluorescence component with lifetimes between 1 and 3 ns. The intensity of the recorded signal fell with increasing distance from the apex, and by 300 nm the signals fell below single-molecule levels for all Si3N4 cantilevers. Silicon cantilevers demonstrated very low count rates relative to single-molecule measurements under all conditions, and virtually no fluorescence. The high density carbon tips also demonstrated low count rates, but the signal contained a short lifetime fluorescence component (0.7 ns). The intensity of the signals from each of the tips was geometry dependent, demonstrating the highest intensities at the edges and corners. Likewise, the anisotropy of all tip signals was observed to be geometry dependent, with the dependence varying on a case-by-case basis. The implications for using confocal illumination instead of total internal reflection are discussed. PMID- 15884086 TI - Two-hybrid fluorescence cross-correlation spectroscopy detects protein-protein interactions in vivo. AB - Fluorescence cross-correlation spectroscopy (FCCS) uses the correlated motion of two distinct fluorophores to detect their interaction. Whereas FCCS has been used with chemically or genetically labeled interaction partners in vitro, FCCS has never been demonstrated in vivo between two autofluorescent proteins. At least one reaction partner was always chemically labeled. Fos and Jun, two components of the AP-1 transcription factor, are known to exert their function as a dimer and can therefore serve as a reference for dimer formation. Expressing fusion proteins between Fos and the enhanced green fluorescent protein (EGFP), as well as Jun and the monomeric red fluorescent protein 1 (mRFP1) in HeLa cells, we show here, for the first time, in vivo FCCS detection of protein-protein interactions. The mobility of the dimerized species is slow, indicating that DNA-binding might stabilize dimerization. The technique has rich potential applications for the rapid screening of protein-protein interactions in vivo, which are able to clarify events during the whole life of cells. PMID- 15884088 TI - Dietary fiber, inflammation, and cardiovascular disease. AB - The role of dietary fiber in the prevention of cardiovascular disease has received increasing attention as data have accumulated. Recent cohort studies have found a consistent protective effect of dietary fiber on cardiovascular disease outcomes, prompting many leading organizations to recommend increased fiber in the daily diet. However, the biologic mechanisms explaining how a fiber influences the cardiovascular system have yet to be fully elucidated. Recent research in large national sample in the USA has demonstrated an association between dietary fiber and levels of C-reactive protein (CRP), a clinical indicator of inflammation. Epidemiologic evidence demonstrating that high-fiber diets are beneficial, coupled with this newer evidence of a possible metabolic effect on inflammatory markers, suggest that inflammation may be an important mediator in the association between dietary fiber and cardiovascular disease (CVD). This paper reviews the evidence for the connections among inflammation, CRP, dietary fiber, and CVD, and recommends further clinical studies using fiber supplementation to isolate and prospectively confirm these important relationships. PMID- 15884089 TI - Determination of non-starch polysaccharides in cereal grains with near-infared reflectance spectroscopy. AB - Cereal grains contain variable amounts of non-starch polysaccharides, such as arabinoxylans and (1-->3),(1-->4)-beta-glucans (beta-glucans), which are associated with their cell walls. The type and composition of these polysaccharides is of increasing interest in both human and animal nutrition. Reference analysis for these polysaccharides requires the use both enzymic and monosaccharide methods. To evaluate fully the non-starch polysaccharides present in grains, some analysts further distinguish between the soluble and insoluble fractions of these components. Near-infrared reflectance (NIR) spectroscopy provides fast, inexpensive analysis. It is, however, a comparative technique that relies on multivariate calibration of sample spectra and accurate reference analysis. It has the potential to be exploited as a rapid analytical method for nutritionally important polysaccharides. The calibration statistics for arabinoxylans and beta-glucans obtained in this study suggest that NIR can be used in plant breeding, nutritional and product studies to obtain simple and rapid estimates of non-starch polysaccharides. The occurrence of wheats with high cell wall contents together with barleys with high beta-glucan contents is well known. However, to date, this genetic variation has not been extensively exploited for the production of grains for use as human food ingredients. PMID- 15884090 TI - Effects of sertraline on depressive symptoms and attentional and executive functions in major depression. AB - Reports on the severity and reversibility of cognitive disturbances in major depression in the literature diverge due to methodological biases. The present study, using a precise methodology, examined attention and executive functions in 20 relatively young, depressed patients presenting a first or second episode of unipolar major depression without psychotic or melancholic characteristics and all being treated with the same psychopharmacological treatment (sertraline) to investigate the changes in potential attentional and executive loss during a subacute period of treatment of 7 weeks. We compared their performance with a group of 26 control subjects who were administered the same cognitive tests. This study confirmed psychomotor slowing associated with attentional and executive disturbance in adults with major depression. Conscious attentional interference for words with a negative emotional valence also was shown. After the first weeks of treatment, the effect of the antidepressant treatment with sertraline was accompanied by a beneficial effect on psychomotor slowing on attentional and executive functions. PMID- 15884091 TI - The natural history of low-grade ductal carcinoma in situ of the breast in women treated by biopsy only revealed over 30 years of long-term follow-up. AB - BACKGROUND: Opportunities to study the natural history of ductal carcinoma in situ (DCIS) are rare. A few studies of incompletely excised lesions in the premammographic era now recognized as DCIS have provided critical insights into its proclivity for local recurrence in the original site. At the time the biopsies in the current study were originally examined, small DCIS was not diagnosed and, by default, these women were treated by biopsy only. METHODS: The authors report the latest results from a follow-up study, which was published originally in 1982, of 28 women with low-grade DCIS who were treated by biopsy only. These women were from a large, prospectively identified, completely characterized cohort. RESULTS: Eleven of 28 women developed invasive breast carcinoma (IBC), all in the same breast and quadrant from which their low-grade DCIS biopsy was taken. Seven IBCs were diagnosed within 10 years of the DCIS biopsy, 1 was diagnosed within 12 years of the DCIS biopsy, and the remaining 3 IBCs were diagnosed over 23-42 years. Five of these women, including 1 woman who developed IBC 29 years after her DCIS biopsy, developed distant metastasis, which resulted in death 1-7 years after the diagnosis of IBC. CONCLUSIONS: The natural history of low-grade DCIS can extend greater than 4 decades, with IBC developing at the same site as the previous DCIS in the majority of women. This natural history differs markedly from that of patients with high-grade DCIS and from the outcome of patients with any completely delimited DCIS excised to negative margins. PMID- 15884092 TI - A three-factor model of the MADRS in major depressive disorder. AB - Major Depressive Disorder (MDD) may be composed of some symptom clusters with distinct neurochemical disturbances, suggesting the importance of the factor analysis of depressive symptoms; however, the results of previous studies using the Montgomery-Asberg Depression Rating Scale (MADRS) have been inconsistent. In the present study, factor analysis of the MADRS was performed in 132 Japanese patients (range 23-74 years, mean 47.6 years) with MDD without any psychiatric comorbidity. The principal component analysis with Varimax rotation identified three factors, accounting for 61% of the total variance: The first factor, labeled dysphoria, included pessimistic thoughts, suicidal thoughts, and reported sadness; the second factor, labeled retardation, included lassitude, inability to feel, apparent sadness, and concentration difficulties; and the third factor, labeled vegetative symptoms, included reduced sleep, reduced appetite, and inner tension. The score of the vegetative factor showed a significant positive correlation with age and was significantly higher in females than in males. This study suggests that the symptoms of MDD, as assessed by the MADRS, cluster into three factors (dysphoria, retardation, and vegetative symptoms). PMID- 15884093 TI - Using patient characteristics and attitudinal data to identify depression treatment preference groups: a latent-class model. AB - A latent-class model is used to identify and characterize groups of patients who share similar attitudes towards treating depression. The results predict the probability of preference-group membership on the basis of observable characteristics and answers to attitudinal questions. Understanding the types of preference groups that exist and a patient's probability of membership in each of the groups can help clinicians tailor the treatment to the patient and may increase patient adherence. One hundred four depressed patients completed a survey on attitudes towards treatment of Major Depressive Disorder. Analysis shows that treatment preferences vary among depressed patients. Three classes are identified that differ in their sensitivity to treatment costs and side effects. One class cares primarily about treatment effectiveness; side effects and the cost of treatment have little impact on this class's treatment decisions. Another class is highly sensitive to cost and side effects. A third class is somewhat sensitive to cost and side effects. Younger and male patients are more likely to be sensitive to treatment costs and side effects. PMID- 15884094 TI - Preserved visual recognition memory in an amnesic patient with hippocampal lesions. AB - There is ongoing debate about whether performance on tests of recognition memory can remain preserved after hippocampal damage. In the present study, we report F.R.G., a patient who became severely amnesic following herpes simplex encephalitis. Although F.R.G. failed all tests involving recall and verbal recognition, she obtained normal performance on a wide number of tests evaluating visual recognition memory (14 of 18 different tests). Her performance was independent of various factors, such as test difficulty, duration of exposure to the stimuli, or delay separating encoding and recognition. F.R.G. also achieved normal performance on two tasks requiring that she associate pairs of visual stimuli. In addition, she demonstrated spared feeling of knowing, suggesting that her performance on recognition tests was explicit and likely to rely on familiarity. Brain imaging (MRI) revealed bilateral lesions of the hippocampus and lesions of the left parahippocampal gyrus, while the right parahippocampal gyrus remained relatively spared. The results of this study support the view that recognition memory can be preserved despite severe hippocampal damage and that familiarity is a distinct memory process that can be dissociated from recollection. PMID- 15884095 TI - Mediotemporal contributions to semantic processing: fMRI evidence from ambiguity processing during semantic context verification. AB - The medial temporal lobe (MTL) is well known to be crucial for various types of memory; however, controversy remains as to which of its substructures contribute to semantic processing and, if so, to what extent. The current study addresses the issue of MTL contributions to semantic processing during lexico-semantic ambiguity processing by using functional magnetic resonance imaging (fMRI) in combination with a context verification task (CVT). The CVT required decisions on the semantic fit of congruent and incongruent target words to the overall meaning of preceding sentential contexts with and without semantic ambiguity. In two of the four experimental conditions (congruent homographic, incongruent homographic), target decisions were critically dependent on the successful processing of prior sentence-final lexico-semantic ambiguity. Semantic context verification per se evidenced bilateral activations of the hippocampus that were part of a functional network including inferior prefrontal and superior parietal cortices. Commonalities in activation differences pertaining to the specific cognitive component of lexico-semantic ambiguity processing were found in a left temporal lobe network that comprised activation foci in the temporal pole, the parahippocampal and fusiform gyri. The present results suggest that the hippocampus may well contribute to semantic processing, namely by a mnemonic function that serves to link the target meaning representation with the meaning of a prior sentence context. Contrary to previous reports from human lesion studies, the present findings further suggest, that the specific cognitive component of lexico-semantic ambiguity processing is neither dependent on the hippocampus nor exclusively subserved by the temporal pole, but also recruits an associative semantic memory function from the parahippocampal gyrus as well as a more general (bottom-up) semantic function from the fusiform gyrus. PMID- 15884096 TI - High-resolution magic-angle-spinning 1H NMR spectroscopy reveals different responses in choline-containing metabolites upon gene therapy-induced programmed cell death in rat brain glioma. AB - Changes in the concentrations of choline-containing metabolites (CCM) have been implicated in both cell proliferation and death processes. In this study, high resolution magic-angle-spinning (HRMAS) 1H NMR spectroscopy was used to study metabolite changes in the CCM chemical shift region in rat glioma ex vivo during apoptosis induced by thymidine kinase-ganciclovir gene therapy. Cell density and apoptotic activity in the tumours were quantified by histological methods. HRMAS 1H NMR was able to resolve peaks from choline (Cho), glycerophosphocholine (GPC), phosphocholine (PC), taurine (Tau) and myo-inositol (myo-Ins), all of which contribute to the in vivo 1H NMR peak centred at 3.23 ppm. The early phase of apoptosis (treatment day 4), with a approximately 2.8-fold increase in the number of apoptotic nuclei (at constant cell density of 1.8 +/- 0.1 x 10(5) cells/mm3) was associated with increases in resonance intensity from GPC and PC, while Cho and Tau remained unchanged. Later stage apoptosis, accompanied by synchronous cell death (cell density declined to 0.7 +/- 0.02 x 10(5) cells/mm3), resulted in a significant decline in Tau relative to untreated tumours, while the contents of CCMs and myo-Ins detectable by 1H HRMAS were unchanged. These observations demonstrate that, while the in vivo 1H NMR peak at 3.23 ppm is indicative of cellular processes involved in apoptosis, the biochemical changes monitored by this resonance involve a number of different and chemically distinct metabolites. PMID- 15884097 TI - Perceptions of insurance coverage for screening mammography among women in need of screening. AB - BACKGROUND: Breast carcinoma remains a significant health problem in the U.S., especially among underserved populations. Although screening mammography is recommended for early detection, in 2002, approximately 25% of women age > 40 years had not had a mammogram within the past 2 years. The current study examined perceptions of insurance coverage and cost as barriers to screening mammography within an underserved, predominantly low-income population of women in need of a mammogram. METHODS: Between 1998 and 2002, face-to-face interviews were conducted with 897 women age > or = 40 years. All women were part of a randomized, controlled study evaluating a health education intervention designed to improve mammography screening. They were asked questions at baseline about cost and insurance coverage as barriers to mammography screening. Women's reports of their level of insurance coverage for mammography were compared with actual coverage by their insurance type to determine the accuracy of their perception of insurance coverage for mammography. The relation between perception of insurance coverage and the barrier of cost was investigated. RESULTS: Greater than half of the women who needed a mammogram identified cost as a barrier to mammography; however, 40% of these women had an inappropriate perception of their insurance coverage. Underestimating or not knowing the level of mammography coverage was strongly associated with reporting cost-related difficulty (odds ratio [OR] = 4.57, 95% confidence interval [95% CI], 1.95-10.70 for the underestimate category; OR = 4.42, 95% CI, 1.80-10.88 for the don't know category), regardless of true coverage levels. CONCLUSIONS: Providing women with information regarding their actual coverage for mammograms may reduce the impact of cost as a barrier to screening mammography. PMID- 15884098 TI - FAWSETS perfusion measurements in exercising skeletal muscle. AB - Arterial spin labeling (ASL) techniques are now recognized as valid tools for providing accurate measurements of cerebral and cardiac perfusion. The labeling process used with most ASL techniques creates two problems, magnetization transfer (MT) effects and arterial transit time effects, that require compensation. The compensation process limits time resolution and hinders absolute quantification. MT effects are particularly problematic in skeletal muscle because they are large and change rapidly during exercise. The protocol presented here was developed specifically for quantification of perfusion in exercising skeletal muscle. The ASL technique that was implemented, FAWSETS, eliminates MT effects and arterial transit times. Localized, single-voxel perfusion measurements were acquired from rat hind limbs at rest, during ischemia and during three different levels of stimulated exercise. The results demonstrate sufficient sensitivity to determine the time constants for perfusion changes at onset of, and during recovery from, exercise and to distinguish the differences in the amplitude of the perfusion response to different levels of exercise. Additional measurements were conducted to demonstrate insensitivity to MT effects. The exercise protocol is easily adaptable to phosphorous magnetic resonance measurements, allowing the possibility to acquire local measurements of perfusion and metabolism from the same tissue in future experiments. PMID- 15884099 TI - Fusion of the EWSR1 and ATF1 genes without expression of the MITF-M transcript in angiomatoid fibrous histiocytoma. AB - Angiomatoid fibrous histiocytoma (AFH) is a rare soft tissue tumor that usually occurs in children and young adults. Only two cases of AFH with genetic rearrangements have been reported previously; both of these had a FUS-ATF1 fusion gene. We have studied an AFH from a 9-year-old boy whose tumor displayed a t(12;22)(q13;q12) as the sole cytogenetic aberration. FISH,RT-PCR, and sequence analyses revealed an EWSR1-ATF1 fusion gene that has previously been reported in clear cell sarcoma (CCS), a soft tissue sarcoma that is morphologically and clinically distinct from AFH. This study thus has demonstrated that the EWSR1 ATF1 chimera represents a fusion gene that can be associated with different tumor types. Simultaneous expression of the EWSR1-ATF1 and MITF-M transcripts in CCS has led to the proposal that the MITF-M promoter is transactivated by EWSR1-ATF1. The AFH, however, did not express the MITF-M transcript, supporting the theory that MITF-M expression in CCS is a reflection of its cellular origin, rather than a consequence of the presence of an EWSR1-ATF1 fusion protein. Activation of the EWSR1-ATF1 oncogene is probably an early step in the transformation process, but the overall gene expression patterns are likely to vary considerably between AFH and CCS, in keeping with their clinicopathologic differences. PMID- 15884100 TI - Breakpoints of variant 9;22 translocations in chronic myeloid leukemia locate preferentially in the CG-richest regions of the genome. AB - From 5% to 10% of 9;22 translocations in chronic myeloid leukemia (CML) are reported to occur in variant form, that is, with the involvement of other regions of the genome in 3-way or more rearrangements. The literature indicates that the alternative breakpoints are not distributed randomly in the genome but show hotspots. We present data on 289 unpublished cases of CML with variant 9;22 translocations having a total of 342 variant breakpoints, the largest independent series to date. We found that the distribution of breaks was in loose agreement with the literature but that some new hotspots were identified; furthermore, some published hotspots were not fully supported by our data. Moreover, when our 342 variant breakpoints were plotted against profiles of CG heterogeneity in the genome, a significant positive correlation between breakpoint locations and CG composition was observed. In an ancillary study, we compared the frequency of variant t(9;22) with that of variants of t(15;17) associated with acute promyelocytic leukemia (AML M3). We found that the frequency of the former, 9.3%, was significantly higher than that of the latter, 2.6%. PMID- 15884101 TI - Short TE single-voxel 1H-MR spectroscopy of hippocampal structures in healthy adults at 1.5 Tesla--how reproducible are the results? AB - The purpose of our study was to evaluate inter- and intra-subject variability and scan-rescan reproducibility of single-voxel 1H-MR spectroscopy (1H-MRS) in hippocampal structures at 1.5 T field strength. Thirty healthy adults were studied bilaterally by optimized, standardized short echo time single-voxel 1H MRS (PRESS, TE=30 ms, TR=3000 ms, oblique voxel orientation, voxel size 2 cm3). Spectral analysis and absolute metabolite quantitation of N-acetylaspartate+N acetylaspartyl-glutamate (tNAA), choline (Cho), creatine (Cr), total glutamate plus glutamine (Glu+Gln) and myo-inositol (Ins) were carried out by LCModel. Inter- and intra-individual reproducibility of these metabolite values were investigated by calculation of mean, standard deviation, coefficient of variation (CV), and by analysis of variance for repeated measurements. The smallest CV in intersubject variability was obtained for tNAA, followed by Cr, Cho, Ins and Glu+Gln. The results of the analysis of variance for repeated measures in inter subject variability showed a marginal effect of scan repetition for Cr (p=0.063) and Glu+Gln (p=0.082); the rescan of both metabolites showed slightly higher concentrations. No statistical significant effect of scan repetition was seen for tNAA (p=0.913), Cho (p=0.857), and Ins (p=0.826). Rescan led to the same results and gave proof of good reproducibility. Scan-rescan testing in one subject showed comparable results: tNAA (CV=4.8%), followed by Cr, Ins, Glu+Gln and Cho (all CV above 10%). PMID- 15884102 TI - Alpha- and beta- aspartyl peptide ester formation via aspartimide ring opening. AB - The undesirable reaction of aspartimide formation has been proved to occur under both acid and base conditions in solid-phase peptide synthesis and is dependent on the beta-carboxyl protecting group, the acid or base used during the synthesis, as well as the peptide sequence. The hydrolysis of aspartimide containing peptides, especially during HPLC purification, yields a mixture of alpha- and beta-aspartyl peptides that can not be purified easily. A previous study demonstrated that treatment of aspartimide-containing peptides with methanol in the presence of 2% diisopropylethylamine in solution leads to alpha- and beta-aspartyl peptide methyl esters. Taking advantage of these results and aiming at elucidating the optimal conditions for aspartimide ring opening, the effect of different types and concentrations of alcohols (primary and secondary) and bases (diisopropylethylamine, collidine, 4-pyrrolidinopyridine, 1-methyl-2 pyrrolidone, piperidine and KCN) was tested at various temperatures and reaction times. The best results were obtained with a combination of a primary alcohol and diisopropylethylamine, while aspartimide ring opening by secondary alcohols occurred only at high temperatures. The optimal conditions were also applied to solid-phase peptide synthesis. PMID- 15884103 TI - MDSIMAID: automatic parameter optimization in fast electrostatic algorithms. AB - MDSIMAID is a recommender system that optimizes parallel Particle Mesh Ewald (PME) and both sequential and parallel multigrid (MG) summation fast electrostatic solvers. MDSIMAID optimizes the running time or parallel scalability of these methods within a given error tolerance. MDSIMAID performs a run time constrained search on the parameter space of each method starting from semiempirical performance models. Recommended parameters are presented to the user. MDSIMAID's optimization of MG leads to configurations that are up to 14 times faster or 17 times more accurate than published recommendations. Optimization of PME can improve its parallel scalability, making it run twice as fast in parallel in our tests. MDSIMAID and its Python source code are accessible through a Web portal located at http://mdsimaid.cse.nd.edu. PMID- 15884104 TI - Conformational studies of oligomeric oxetane-based dipeptide isosteres derived from L-rhamnose or D-xylose. AB - Conformational investigations have been undertaken on oligomers (dimers, tetramers, hexamers) of five closely related oxetane-based dipeptide isosteres. All the oligomers were subjected to a range of studies by NMR, FT-IR and CD spectroscopy. The oligomers derived from methyl 2,4-anhydro-5-azido-3-O-tert butyldimethylsilyl-5-deoxy-L-rhamnonate 'monomer' all exhibited evidence of ordered conformations in chloroform and 2,2,2-trifluoroethanol (TFE) solution. 5 Acetamido and N-methylamide derivatives of the L-rhamnonate 'monomer', along with a 'dimer' lacking silyl protection at C-3, were synthesized to ascertain the role of intramolecular interactions. This led to the conclusion that, for the L rhamnonate oligomers, steric interactions govern the conformational preference observed. The equivalent silyl-protected D-lyxonate oligomers gave ordered CD spectra in TFE solution, but NMR and FT-IR spectroscopy in chloroform solution suggested an irregular, non-hydrogen bonded system. The remaining silyl-protected 6-deoxy-L-altronate, 6-deoxy-D-gulonate and D-fuconate oligomers appear to be characterized by their lack of ordered conformation in TFE and chloroform solution. PMID- 15884105 TI - Normal aspects of colorectal motility and abnormalities in slow transit constipation. AB - Human colonic motility is a relatively difficult topic to investigate. However, the refinement of manometric techniques in recent years enabled us to study both the proximal and distal segments of the viscus. The present paper reviews our knowledge about normal aspects of colorectal motility in man and the abnormalities found in slow transit constipation (STC), one of the most frequent and difficult to treat subtypes of constipation. An internet-based search strategy of the Medline and Science Citation Index was performed using the keywords colon, colonic, colorectal, constipation, slow transit, motility, rectal, rectum in various combinations with the Boolean operators AND, OR and NOT. Only articles related to human studies were used, and manual cross referencing was also performed. Most of colonic motor activity is represented by single nonpropagated contractions, rarely organized in bursts; this activity is maximal during the day, especially after waking and following meals. In addition, a specialized propagated activity with propulsive features is detectable, represented by high- and low-amplitude propagated contractions. In the severe form of constipation represented by the slow transit type, the above motor activity is completely deranged. In fact, both basal segmental activity (especially in response to meals) and propagated activity (especially that of high amplitude) are usually decreased, and this may represent a physiologic marker of this disorder. Human colonic motor activity is quite a complex issue, still only partly understood and investigated, due to anatomic and physiological difficulties. In recent years, however, some more data have been obtained, even in proximal segments. These data have helped in elucidating, although only in part, some pathophysiological mechanisms of chronic constipation, and especially of the STC subtype. PMID- 15884106 TI - Free radicals and antioxidant systems in reflux esophagitis and Barrett's esophagus. AB - AIM: Experimental studies suggest that free radicals are involved in acid and pepsin-induced damage of esophageal mucosa. The profile and balance between free radicals and antioxidant systems in human esophagitis are unknown. METHODS: Superoxide anion and its powerful oxidant reaction with nitric oxide (peroxynitrite) generation were determined in esophageal mucosal biopsies from 101 patients with different gastro-esophageal reflux diseases and 28 controls. Activity of both superoxide dismutase (SOD) and catalase, and reduced glutathione (GSH) levels, were also assessed. Expression of Cu, ZnSOD, MnSOD and tyrosine nitrated MnSOD were analyzed by Western blot and/or immunohistochemistry. RESULTS: The highest levels of superoxide anion generation were found in patients with severe lesions of esophagitis. Peroxynitrite generation was intense in Barrett's biopsies, weaker in esophagitis and absent/weak in normal mucosa. Expression of Cu, ZnSOD and MnSOD isoforms were present in normal mucosa and increased according to the severity of the lesion, reaching the highest level in Barrett's esophagus. However, SOD mucosal activity significantly decreased in patients with esophagitis and Barrett's esophagus, which was, at least in part, due to nitration of its tyrosine residues. Catalase activity and GSH levels were significantly increased in mucosal specimens from patients with esophagitis and/or Barrett's esophagus. CONCLUSION: A decrease in SOD antioxidant activity leading to increased mucosal levels of superoxide anion and peroxynitrite radicals may contribute to the development of esophageal damage and Barrett's esophagus in patients with gastroesophageal reflux. Administration of SOD may be a therapeutic target in the treatment of patients with esophagitis and Barrett's esophagus. PMID- 15884107 TI - Expression of lysosome-associated protein transmembrane 4B-35 in cancer and its correlation with the differentiation status of hepatocellular carcinoma. AB - AIM: To produce high-quality polyclonal antibody to lysosome-associated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellular carcinoma (HCC). METHODS: The 297 bp 5' end of LAPTM4B cDNA was obtained by PCR and inserted into prokaryotic expression vector pGEX-KG. Then the recombinant pGEX-KG-N(1-99) was transformed into E.coli JM109 to express GST-fusion protein. The fusion protein was purified by glutathione sepharose(TM) 4B agarose. The purified GST-LAPTM4B N(1-99) was characterized by SDS-PAGE, and used to immunize rabbits. The titer and specificity of antisera were detected by ELISA and Western blot, respectively. The correlation between the expression levels of LAPTM4B-35 and the differentiation status of HCC was analyzed via Western blot. The expression of LAPTM4B-35 in HCC and other six cancer tissues was investigated via tissue chip and immunohistochemical analysis. RESULTS: About 6.2 mg of pure GST-LAPTM4B-N(1 99) was isolated from 1 L of bacteria. The GST-LAPTM4B-N(1-99) produced high titer antisera in rabbits and showed good immunity. Western blot showed specific reactions for the antibody to the LAPTM4B-35 in the total proteins from HCC tissues and BEL-7402 cells, also to the fusion protein purified or in the transformed bacteria. LAPTM4B-35 was remarkably expressed in several cancers, such as HCC, breast cancer, gastric carcinoma, lung cancer, and colon carcinoma, but not commonly expressed in esophageal cancer and rectum carcinoma. Notably, the expression levels of LAPTM4B-35 were significantly and inversely correlated to the differentiation of HCCs in a 20 case analysis. CONCLUSION: Specific polyclonal antibody (LAPTM4B-N(1-99)-pAb) to LAPTM4B-35 was produced. It identified the expression of LAPTM4B-35 in some cancer tissues originated from single layer cuboidal and columnar epithelial cells and firmly demonstrated that the expression of LAPTM4B-35 in HCC was inversely correlated with the differentiation of HCC. PMID- 15884108 TI - Expression and clinical significance of TAp73alpha, p53, PCNA and apoptosis in hepatocellular carcinoma. AB - AIM: To study the prognostic role of TAp73alpha, p53, proliferating cell nuclear antigen (PCNA) and apoptosis in patients with hepatocellular carcinoma (HCC) after surgical tumor ablation. METHODS: Forty-seven human resected HCC tissues and 42 adjacent non-cancerous tissues were studied with 10 normal liver tissues as control group. TAp73alpha, p53, and PCNA were detected with Elivision immunohistochemistry. Terminal deoxynucleotidyl transferase (TdT)-mediated d-UTP biotin nick-end labeling (TUNEL) method was used to detect the apoptosis cells. All clinical and pathological materials were analyzed by SPSS10.0 statistical package. RESULTS: TAp73alpha overexpressed in HCC tissues (36.2%) when compared with adjacent non-cancerous tissues (2.38%, P<0.005) and normal liver tissues (0, P<0.01). Mutant type p53 (mt-p53) overexpressed in HCC tissues (38.3%) when contracted with adjacent non-cancerous tissues (16.7%, P<0.05) and normal liver tissues (0, P<0.01). Proliferation index (PI) level in HCC tissues was significantly higher than that in adjacent non-cancerous tissues (30.34%+/-4.46% vs 27.88%+/-5.89%, t, P = 0.028). Apoptosis index (AI) level in HCC tissues was higher than that in adjacent non-cancerous tissues (8.62%+/-2.28% vs 7.38%+/ 2.61%, t, P = 0.019). Expression of TAp73alpha was associated with lymph node metastasis and mt-p53, with r = 0.407 and 0.265, respectively. Expression of mt p53 was associated with Edmondson's stage and AFP, with r = 0.295 and -0.357, respectively. In Kaplan-Meier univariant analysis, TAp73alpha, AFP, TNM stage, portal vein invasion, liver membrane invasion and HBsAg correlated with prognosis (log rank, P = 0.039, 0.012, 0.002, 0.000, 0.014, 0.007, respectively). Multivariant Cox regression analysis showed that TAp73alpha, AFP, TNM stage, portal vein invasion, liver membrane invasion and age were independent factors of prognosis. CONCLUSION: These results suggest that TAp73alpha can be used as a prognostic indicator of patients with HCC undergoing surgical tumor ablation. AFP, TNM, portal vein invasion, liver membrane invasion and age also have a potency of predicting the prognosis of HCC. PMID- 15884109 TI - YMDD variants of HBV DNA polymerase gene: rapid detection and clinicopathological analysis with long-term lamivudine therapy after liver transplantation. AB - AIM: To look for a rapid low-cost technique for the detection of HBV variants. METHODS: Two patients who underwent orthotopic liver transplantation (OLT) for HBV infection were treated with lamivudine (100 mg daily) and HBV infection recurred in the grafted livers. The patients were monitored intensively for liver enzymes, hepatitis B surface antigen (HBsAg) and HBV DNA in serum. Liver biopsy was performed regularly. HBV DNA in a conserved polymerase domain (the YMDD locus) was amplified from serum of each patient by PCR and sequenced. HBV genotypes were analyzed by restriction fragment length polymorphism (RFLP) of the PCR products generated from a fragment of the polymerase gene. RESULTS: YMDD wild type HBV was detected in one patient by PCR-RFLP and DNA sequencing 19 mo after OLT, and YIDD mutant-type HBV in the other patient, 16 mo after OLT. CONCLUSION: PCR-RFLP assay is an accurate and simple method for genotyping lamivudine resistant HBV variants. PMID- 15884110 TI - Increased activity of Pgp multidrug transporter in patients with Helicobacter pylori infection. AB - AIM: To determine whether local antibiotic resistance involves P-glycoprotein (Pgp)-mediated active drug out-pumping during Helicobacter pylori (H pylori) infection treatment with classic antibiotic therapy. METHODS: Pgp activity was determined in gastric mucosa biopsy specimens obtained from 53 patients with pathohistologically verified gastritis and microbiologically confirmed H pylori infection, and compared with the Pgp activity in 12 control subjects with normal endoscopic findings. The H pylori positive patients were treated with short-term 7-d therapy consisting of two antibiotics (amoxicillin and azithromycin/metronidazole and clarithromycin) and a proton pump inhibitor. Pgp activity was determined by flow cytometry in the test of rhodamine dye efflux and quantified as mean fluorescence ratio (RMF). RESULTS: Upon the first cycle, H pylori was successfully eradicated in 20 patients, whereas therapy was continued in 33 patients. In the course of antibiotic therapy, RMF increased (P<0.05) and gastric cells showed higher rhodamine dye efflux. The mean pre-treatment RMF values were also higher (P<0.0001) in patients with multiple therapeutic failure than in those with successful H pylori eradication and control subjects. CONCLUSION: Pgp might be one of the causes of therapy failure in patients with H pylori and antibiotic therapy could be chosen and followed up on the basis of the Pgp transporter local activity. PMID- 15884111 TI - Do patients with non-ulcer dyspepsia respond differently to Helicobacter pylori eradication treatments from those with peptic ulcer disease? A systematic review. AB - AIM: It is controversial whether patients with non-ulcer dyspepsia (NUD) respond differently to Helicobacter pylori (H pylori) eradication treatment than those with peptic ulcer disease (PUD). To review the evidence for any difference in H pylori eradication rates between PUD and NUD patients. METHODS: A literature search for full articles and meeting abstracts to July 2004 was conducted. We included studies evaluating the efficacy of a proton pump inhibitor (P) or ranitidine bismuth citrate (RBC) plus two antibiotics of clarithromycin (C), amoxicillin (A), metronidazole (M), or P-based quadruple therapies for eradicating the infection. RESULTS: Twenty-two studies met the criteria. No significant difference in eradication rates was found between PUD and NUD patients when treated with 7-d RBCCA, 10-d PCA or P-based quadruple therapies. When the 7-d PCA was used, the pooled H pylori eradication rate was 82.1% (431/525) and 72.6% (448/617) for PUD and NUD patients, respectively, yielding a RR of 1.15 (95%CI 1.01-1.29). However, the statistically significant difference was seen only in meeting abstracts, but not in full publications. CONCLUSION: There is no convincing evidence to suggest that NUD patients respond to H pylori eradication treatments differently from those with PUD, although a trend exists with the 7-d PCA therapy. PMID- 15884112 TI - Effects of mycophenolate mofetil vs cyclosporine administration on graft survival and function after islet allotransplantation in diabetic rats. AB - AIM: To develop an experimental model of islet allotransplantation in diabetic rats and to determine the positive or adverse effects of MMF as a single agent. METHODS: Thirty-six male Wistar rats and 18 male Lewis rats were used as recipients and donors respectively. Diabetes was induced by the use of streptozotocin (60 mg/kg) intraperitoneally. Unpurified islets were isolated using the collagenase digestion technique and transplanted into the splenic parenchyma. The recipients were randomly assigned to one of the following three groups: group A (control group) had no immunosuppression; group B received cyclosporine (CsA) (5 mg/kg); group C received mycophenolate mofetil (MMF) (20 mg/kg). The animals were killed on the 12th d. Blood and grafted tissues were obtained for laboratory and histological assessment. RESULTS: Median allograft survival was significantly higher in the two therapy groups than that in the controls (10 and 12 d for CsA and MMF respectively vs 0 d for the control group, P<0.01). No difference in allograft survival between the CsA and MMF groups was found. However, MMF had less renal and hepatic toxicity and allowed weight gain. CONCLUSION: Monotherapy with MMF for immunosuppression was safe in an experimental model of islet allotransplantation and was equally effective with cyclosporine, with less toxicity. PMID- 15884113 TI - Protective role of metallothionein in stress-induced gastric ulcer in rats. AB - AIM: To illustrate the pathophysiological role of metallothionein (MT) in gastric ulcer induced by stress. METHODS: Wistar rats underwent water-immersion-restraint (WIR) stress, ZnSO(4) (an MT inducer) treatment, WIR+ZnSO(4) or WIR+MT, and the ulcer index (UI) was estimated in excised stomach and liver tissues. The mRNA level of gastric MT was determined by semi-quantitative RT-PCR. The MT content in gastric and hepatic tissues was determined by Cd/hemoglobin affinity assay. The lipid peroxidation products malondialdehyde (MDA) and conjugated dienes (CD) were estimated by use of thiobarbituric acid reactive species and ultraviolet spectrophotometry. RESULTS: WIR stress induced severe gastric mucosal lesions in rats. Compared with control rats, stressed rats had increased lipid peroxide content in serum and stomach and liver tissues. MDA content was increased by 34%, 21% and 29% and CD level by 270%, 83% and 28%, respectively. MT content in the stomach and liver was increased by 0.74- and 1.8-fold, and the MT-mRNA level in the stomach was increased by 26%. Pretreatment with ZnSO(4) prevented gastric lesion development (the UI was 87% lower than that without pretreatment), and the MDA and CD content in serum and tissues was lower. The MT content in the liver was double in rats that were not pretreated, and the MT mRNA level in the stomach was 35% higher. MT administration 1 h before the WIR stress prevented gastric lesion development (the UI decreased by 47% compared with that in rats not pretreated), and the MDA and CD content in serum and tissues was significantly lower. CONCLUSION: In WIR-stressed rats, the MT level was increased in serum and in stomach and liver tissues. Pre-administration of exogenous MT or pre-induction of endogenous MT can protect the gastric mucosa against stress-induced ulcers and inhibits the formation of stress-induced lipid peroxide. MT could have a gastroprotective effect and might be a new interventive and therapeutic target in stress-induced gastric ulcers. PMID- 15884114 TI - Clinicopathological significance of Bcl-2 and Bax protein expression in human pancreatic cancer. AB - AIM: To assess the clinicopathological significance of the expression of the apoptosis-inhibitory Bcl-2 protein (pBcl-2) and the apoptosis-promoting Bax protein (pBax) in human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: Fifty-nine surgical specimens of IDCs of the pancreas were stained immunohistochemically to detect pBcl-2 and pBax expressions whose correlation to tumor classification, staging, and prognosis was analyzed by univariate and multivariate analyses. RESULTS: The expression of pBcl-2 and pBax was detected in 21 of 59 (35.6%) and in 29 of 59 (49.2%) patients with IDCs of the pancreas, respectively. Neither pBcl-2 nor pBax alone was correlated to TNM staging and differentiation degree of IDCs of the pancreas according to univariate analysis. By Mantel-Cox test, the median survival time after surgery for pBcl-2(+) and pBcl 2(-) groups were 14.3 and 7.3 mo, respectively (chi(2) = 9.357, P = 0.002) and that for pBax(+) and pBax(-) groups were 12.9 and 10.2 mo, respectively (chi(2) = 0.285, P>0.05). Contingency coefficient between pBcl-2 and pBax expression was 0.298, indicating that there was correlation between them (chi(2) = 5.74, P<0.05). The median survival time after surgery for pBcl-2(+)pBax(+) and pBcl 2(+)pBax(-) groups were 14.3 and 14.1 mo, respectively, and that for pBcl-2( )pBax(+) and pBcl-2(-)pBax(-) groups were 5.9 and 9.9 mo, respectively. There was a significant difference between pBcl-2(+)pBax(+) and pBcl-2(-)pBax(+) (chi(2) = 5.06, P<0.05), such was the case for pBcl-2(+)pBax(+) and pBcl-2(-)pBax(-) (chi(2) = 7.18, P<0.01). Cox proportional hazards model for multivariate analysis was applied, indicating that pBcl-2, TNM staging, age and pBax were high risk factors of post-surgical survival time. CONCLUSION: Both pBcl-2 and pBax have high expression in IDCs of the pancreas, indicating that co-expression of pBcl-2 and pBax is a good indicator of favorable prognosis in IDCs of the pancreas. PMID- 15884115 TI - Drug sensitivity and drug resistance profiles of human intrahepatic cholangiocarcinoma cell lines. AB - AIM: To study the effect of a number of chemotherapeutic drugs on five human intrahepatic cholangiocarcinoma (CCA) cell lines. The expressions of genes that have been proposed to influence the resistance of chemotherapeutic drugs including thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), glutathione-S-transferase P1 (GSTP1), multidrug resistance protein (MDR1) and multidrug resistance-associated proteins (MRPs) were also determined. METHODS: Five human CCA cell lines (KKU-100, KKU-M055, KKU-M156, KKU-M214 and KKU-OCA17) were treated with various chemotherapeutic drugs and growth inhibition was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4 sulfophenyl)-2H-tetrazolium (MTS) assay. Semi-quantitative levels of gene expression were determined by a reverse transcriptase polymerase chain reaction (RT-PCR). Results of IC(50) values and the ratios of gene expression were analyzed by linear regression to predict their relationship. RESULTS: Among five CCA cell lines, KKU-M055 was the most sensitive cell line towards all chemotherapeutic drugs investigated, particularly taxane derivatives with IC(50) values of 0.02-3 nmol/L, whereas KKU-100 was apparently the least sensitive cell line. When compared to other chemotherapeutic agents, doxorubicin and pirarubicin showed the lowest IC(50) values (<5 mumol/L) in all five CCA cell lines. Results from RT-PCR showed that TS, MRP1, MRP3 and GSTP1 were highly expressed in these five CCA cell lines while DPD and MRP2 were only moderately expressed. It should be noted that MDR1 expression was detected only in KKU-OCA17 cell lines. A strong correlation was only found between the level of MRP3 expression and the IC(50) values of etoposide, doxorubicin and pirarubicin (r = 0.86-0.98, P<0.05). CONCLUSION: Sensitivity to chemotherapeutic agents is not associated with the histological type of CCA. Choosing of the appropriate chemotherapeutic regimen for the treatment of CCA requires knowledge of drug sensitivity. MRP3 was correlated with resistance of CCA cell lines to etoposide, doxorubicin and pirarubicin, whereas other chemotherapeutic drugs showed no association. The role of this multidrug resistance-associated protein, MRP3, in chemotherapeutic resistance in CCA patients needs to be further investigated. PMID- 15884116 TI - Blood platelet and monocyte activations and relation to stages of liver cirrhosis. AB - AIM: Blood platelets (plt) and monocytes are the cells that play a crucial role in the pathogenesis of liver damage and liver cirrhosis (LC). In this paper, the analysis of mutual relationship between platelets and monocytes activation in LC was conducted. METHODS: Immunofluorescent flow cytometry was used to measure the percentage of activated platelet populations (CD62P, CD63), the percentage of plt monocyte aggregates (pma) (CD41/CD45), and activated monocytes (CD11b, CD14, CD16) in the blood of 20 volunteers and 40 patients with LC. Platelet activation markers: sP-selectin, platelet factor 4 (PF4), beta-thromboglobulin (betaTG) and monocyte chemotactic peptide-1 (MCP-1) were measured and compared in different stages of LC. RESULTS: Platelet activation with the increase in both betaTG serum concentration and elevation of plt population (CD62P and CD63 as well as MIF CD62P and CD63) is elevated as LC develops and thrombocytopenia rises. There is a positive correlation between medial intensity of fluorescence (MIF) CD62P and MIF CD63 in LC. We did not show any relationship between monocyte activation and pma level. SP-selectin concentration correlates positively with plt count and pma, and negatively with stage of plt activation and MIF CD62P and MIF CD63. There was no correlation between MCP-1 concentration and plt, monocyte activation as well as pma level in LC. CD16 monocytes and MIF CD16 populations are significantly higher in the end stage of LC. A positive correlation occurs between the value of CD11b monocyte population and MIF CD14 and MIF CD16 on monocytes in LC. CONCLUSION: Platelet and monocyte activation plays an important role in LC. Platelet activation stage does not influence monocyte activation and production of plt aggregates with monocytes in LC. With LC development, thrombocytopenia may be the result of plt consumption in platelet-monocyte aggregates. PMID- 15884117 TI - Association of Helicobacter pylori infection and giardiasis: results from a study of surrogate markers for fecal exposure among children. AB - AIM: To investigate whether Helicobacter pylori (H pylori) infection is associated with hepatitis A virus (HAV) infection, presence of enteroparasites, and other surrogates of fecal exposure. METHODS: We conducted a cross-sectional study in 121 children consecutively admitted at a pediatric hospital in Salvador, Brazil. H pylori and HAV infection were identified by the presence of serum antibodies. Stool specimens were examined for the presence of ova and parasites. A structured questionnaire inquiring about sanitary conditions and life style was applied to each subject. RESULTS: Fifty-one of the 121 children (42.1%) were found to be seropositive for H pylori, and 45 (37.2%) for HAV. The seroprevalence of H pylori and HAV both increased significantly with age. Cross-tabulation of data showed that 26 (21.5%) were seropositive and 51 (42.1%) were negative for both H pylori and HAV antibodies (chi(2) = 7.18, OR = 2.8, CI 1.30-5.97). The age adjusted OR for an HAV-infected child being H pylori positive was 2.3 (CI 1.02 5.03). The agreement between H pylori and HAV seropositivity was fair (kappa = 0.24). After controlling for possible confounding, the variables remaining independently associated with seropositivity to H pylori were age, presence of Giardia lamblia in feces (OR = 3.2, 95%CI, 1.1-9.5) and poor garbage disposal quality (OR = 2.4, 95%CI, 1.1-5.1). CONCLUSION: Our data suggest that H pylori infection is associated with surrogate markers of fecal exposure. Thus, we conclude that the fecal-oral route is relevant in the transmission of HP among children in an urban setting of a developing country. The association observed between G. lamblia and H pylori infection may have several explanations. Further studies to investigate this relationship are warranted. PMID- 15884118 TI - Helicobacter pylori eradication lowers serum homocysteine level in patients without gastric atrophy. AB - AIM: To determine whether Helicobacter pylori (H pylori)infection caused hyperhomocysteinemia by altering serum vitamin B(12), serum folate and erythrocyte folate levels and whether eradication of this organism decreased serum homocysteine level. METHODS: The study involved 73 dyspeptic H pylori positive patients, none of them had gastric mucosal atrophy based on rapid urease test and histology. Out of 73 patients, 41 (56.2%) showed a successful eradication of H pylori 4 wk after the end of treatment. In these 41 patients, fasting serum vitamin B(12), folate and homocysteine levels, and erythrocyte folate levels before and 4 wk after H pylori eradication therapy were compared. RESULTS: The group with a successful eradication of H pylori had significantly higher serum vitamin B(12) and erythrocyte folate levels in the post-treatment period compared to those in pre-treatment period (210+/-97 pg/mL vs 237+/-94 pg/mL, P<0.001 and 442+/-212 ng/mL vs 539+/-304 ng/mL, P = 0.024, respectively), but showed no significant change in serum folate levels (5.6+/-2.6 ng/mL vs 6.0+/ 2.4 ng/mL, P = 0.341). Also, the serum homocysteine levels in this group were significantly lower after therapy (13.1+/-5.2 micromol/L vs 11.9+/-6.2 micromol /L, P = 0.002). Regression analysis showed that serum homocysteine level was positively correlated with age (P = 0.01) and negatively with serum folate level before therapy (P = 0.003). CONCLUSION: Eradication of H pylori decreases serum homocysteine even in patients who do not exhibit gastric mucosal atrophy. It appears that the level of homocysteine in serum is related to a complex interaction among serum vitamin B(12), serum folate and erythrocyte folate levels. PMID- 15884119 TI - Genetic association of cytokines polymorphisms with autoimmune hepatitis and primary biliary cirrhosis in the Chinese. AB - AIM: To characterize gene polymorphism of several cytokine gene in-patients with AIH and PBC and to analyze the difference of the polymorphism distribution between Chinese patients and healthy controls. METHODS: The study population consisted of 62 patients with AIH, and 77 patients with PBC. The genetic profile of four cytokines was analyzed by restriction fragment length polymorphism after specific PCR amplification (PCR-RFLP) or sequence-specific primers PCR (SSP-PCR). The analyzed gene polymorphism included interleukin-1 (IL-1) (at position +3 953 and IL-1RN intron 2), IL-6 (at position -174), IL-10 promoter (at position -1 082, -819, and -592). The control group consisted of 160 healthy blood donors. RESULTS: The majority of Chinese people including patients and healthy controls exhibited IL-1B 1,1 genotype, and there was no significant difference in AIH, PBC patients and controls. There were highly statistically significant differences in the distribution of the IL-1RN gene polymorphism between the patients with PBC compared with controls. The frequency of IL-1RN 1,1 was significantly higher (90.9% vs 79.4%, P = 0.03) and the frequency of IL-1RN 1,2 was significantly lower in PBC patients (6.5% vs 17.5%, P = 0.01). No statistical difference was observed between AIH patients and controls. All of the 160 healthy controls and 62 cases of AIH patients exhibited IL-6-174GG genotype, and there were four cases, which expressed IL-6-174GC genotype in 77 cases of PBC patients. The frequency of IL-6-174GC was markedly significantly higher in PBC patients compared with controls (5.2% vs 0%, P = 0.004). No statistically significant difference was found in the distribution of IL-10 promoter genotype in AIH and PBC patients compared with controls. CONCLUSION: The polymorphisms of IL-1RN and IL-6 -174G/C appear to be associated with PBC in Chinese patients. PMID- 15884120 TI - Assessment of small intestinal bacterial overgrowth in uncomplicated acute diverticulitis of the colon. AB - AIM: Small intestinal bacterial overgrowth (SIBO) may contribute to the appearance of several gastrointestinal nonspecific symptoms. Acute diverticulitis is affected by some similar symptoms and bacterial colonic overgrowth. We assessed the prevalence of SIBO in acute uncomplicated diverticulitis and evaluated its influence on the clinical course of the disease. METHODS: We studied 90 consecutive patients (39 males, 51 females, mean age 67.2 years, range 32-91 years). Sixty-one patients (67.78%) and 29 patients (32.22%) were affected by constipation-or diarrhea-prevalent diverticulitis respectively. All subjects were investigated by lactulose H2-breath test at the entry and at the end of treatment. We also studied a control group of 20 healthy subjects (13 males, 7 females, mean age 53 years, range 22-71 years). RESULTS: Oro-cecal transit time (OCTT) was delayed in 67/90 patients (74.44%) (range 115-210 min, mean 120 min). Fifty-three of ninety patients (58.88%) showed SIBO, while OCTT was normal in 23/90 patients (25, 56%). In the control group, the mean OCTT was 88.2 min (range 75-135 min). The difference between diverticulitic patients and healthy subjects was statistically significant (P<0.01). OCTT was longer in constipation-prevalent disease than in diarrhea-prevalent disease (180.7 min (range 150-210 min) vs 121 min (range 75-180 min) (P<0.001)), but no difference in bacterial overgrowth was found between the two forms of diverticulitis. After treatment with rifaximin plus mesalazine for 10 d, followed by mesalazine alone for 8 wk, 70 patients (81.49%) were completely asymptomatic, while 16 patients (18.60%) showed only slight symptoms. Two patients (2.22%) had recurrence of diverticulitis, and two other patients (2.22%) were withdrawn from the study due to side-effects. Seventy nine of eighty-six patients (91.86%) showed normal OCTT (range 75-105 min, mean 83 min), while OCTT was longer, but it was shorter in the remaining seven (8.14%) patients (range 105-115 min, mean of 110 min). SIBO was eradicated in all patients, while it persisted in one patient with recurrence of diverticulitis. CONCLUSION: SIBO affects most of the patients with acute diverticulitis. SIBO may worsen the symptoms of patients and prolong the clinical course of the disease, as confirmed in the case of persistence of SIBO and diverticulitis recurrence. In this case, we can hypothesize that bacteria from small bowel may re-colonize in the colon and provoke recurrence of symptoms. PMID- 15884121 TI - Diagnostic value of fine motor deficits in patients with low-grade hepatic encephalopathy. AB - AIM: The role of motor dysfunction in early diagnosis of low-grade hepatic encephalopathy remains uncertain. We performed a pilot study to comparatively investigate the kinematic characteristics of small and large rapid alternating movements in patients with liver cirrhosis and low-grade hepatic encephalopathy. METHODS: A kinematic analysis of alternating handwriting (7.5 mm) and large drawing movements (DM, 175 mm) was performed in 30 patients with liver cirrhosis (no hepatic encephalopathy: n = 10; minimal hepatic encephalopathy: n = 9; grade I hepatic encephalopathy: n = 11; healthy controls: n = 12). The correlation between kinematic parameters, clinical neuro-psychiatric symptoms of cerebral dysfunction and the grade of encephalopathy was investigated. RESULTS: Both movement types, handwriting and drawing, were significantly slower in cirrhotic patients. In contrast to large DM, the deterioration of handwriting movements significantly correlated with the increase of symptoms of motor dysfunction and differentiated significantly within the group of cirrhosis patients corresponding to the degree of hepatic encephalopathy. CONCLUSION: The deterioration of fine motor control is an important symptom of low-grade hepatic encephalopathy. The kinematic analysis of handwriting allows the quantitative analysis of alterations of motor function and is a possible tool for diagnostics and monitoring of motor dysfunction in patients with low-grade hepatic encephalopathy. PMID- 15884122 TI - Dinucleotide microsatellite repeats are essential for the diagnosis of microsatellite instability in colorectal cancer in Asian patients. AB - AIM: The molecular diagnosis of microsatellite instability (MSI) in colorectal cancer (CRC) is based on the analysis of five microsatellite markers. Among them, the two mononucleotide microsatellite repeats are considered more informative for this analysis than the three dinucleotide ones. The aim of this study is to establish the most relevant markers for MSI analysis in colorectal cancers from Asian patients. METHODS: The MSI analysis of 143 CRC cases in a routine molecular diagnostic laboratory was reviewed. Analysis by fluorescence-based PCR of the five recommended microsatellites was performed, followed by data interpretation according to internationally accepted guidelines. The results were analyzed to address (1) the rate of success in the analysis of histopathological samples not specifically prepared for molecular analysis; (2) the relative importance of individual markers in the diagnosis of high-MSI (H-MSI). RESULTS: MSI analysis was unsuccessful in 34 cases (24%), but for tissues archived in recent years the unsuccessful rate was 5%. We found the D2S123 marker the most vulnerable to inadequate tissue preservation, failing to amplify in 58 instances. Approximately 30% (32/109) of the cases were H-MSI, while 7/109 (6%) were low-MSI. A detailed analysis of the H-MSI cases revealed that the dinucleotide repeats (and D5S346 in particular) were more relevant than the mononucleotide repeats in assigning the correct MSI status. CONCLUSION: The analysis of dinucleotide repeats is essential for the establishment of MSI status in Asian CRC patients. PMID- 15884123 TI - Antral exfoliative cytology for the detection of Helicobacter pylori in the stomach. AB - AIM: To evaluate the efficacy of antral exfoliative cytology method in the diagnosis of Helicobacter pylori (H pylori) infection in the stomach. METHODS: Fifty patients were submitted to upper digestive tract endoscopy due to complaints of dyspepsia. The material for exfoliative cytology was obtained by extensive brushing of the gastric antral mucosa and Papanicolaou stain was used to identify the bacteria. The authors also performed gastric biopsies to collect material for urease tests and histologic studies, with hematoxylin-eosin and fucsin stains in order to identify the microorganism. The gold standard used to detect the presence of H pylori was an analysis of the combined results from the gastric biopsies by urease test and histological method. RESULTS: Antral exfoliative cytology method exhibited 90.3% sensitivity, 66.6% specificity, accuracy of 81.6%, positive predictive value of 82.3% and negative predictive value of 80.0%, in this population with a prevalence of 63.3%. CONCLUSION: Antral exfoliative cytology was demonstrated to be a sensitive, accurate and easy to perform method for investigating H pylori infection in the stomach. PMID- 15884124 TI - Dextrose in the banked blood products does not seem to affect the blood glucose levels in patients undergoing liver transplantation. AB - AIM: Hyperglycemia commonly seen in liver transplantation (LT) has often been attributed to the dextrose in the storage solution of blood transfusion products. The purpose of the study is to compare the changes of the blood glucose levels in transfused and non-transfused patients during LT. METHODS: A retrospective study on 60 biliary pediatric patients and 16 adult patients undergoing LT was carried out. Transfused pediatric patients were included in Group I (GI), those not transfused in Group II (GII). Twelve adult patients were not given transfusion and assigned to Group III (GIII); whereas, four adult patients who received massive transfusion were assigned to Group IV (GIV). The blood glucose levels, volume of blood transfused, and the volume of crystalloid infused were recorded, compared and analyzed. RESULTS: Results showed that the changes in blood glucose levels during LT for both non-transfused and minimally transfused pediatric groups and non-transfused and massively-transfused adult groups were almost the same. CONCLUSION: We conclude that blood transfusion does not cause significant changes in the blood glucose levels in this study. PMID- 15884125 TI - Survival outcome of lobar or segmental transcatheter arterial embolization with ethanol-lipiodol mixture in treating hepatocellular carcinoma. AB - AIM: To evaluate the clinical outcome and cost-effectiveness of transcatheter arterial ethanol-lipiodol embolotherapy on hepatocellular carcinoma (HCC). METHODS: One hundred patients with HCC who were treated only by lobar or segmental transarterial embolization (TAE) with ethanol-lipiodol mixture were enrolled in this study. The 1st- and 2nd-year survival rates were analyzed to evaluate the feasibility of its method. These outcomes of our patients were individually correlated to the Child-Pugh classification and the computed tomographic features of HCC. RESULTS: The overall 1st- and 2nd-year survival rates were 72% and 46%, respectively. The patients were classified into three groups according to their liver function status: 68 patients as Child-Pugh class A, 26 as Child B, and 6 as Child C. Child A had better survival rate than the Child B and/or C. The 1st-year survival rates of patients with Child A-C were 84%, 50%, and 33.3% respectively and the 2nd-year survival rates were 55.5%, 28.5%, and 33.3%, respectively. According to the computed tomographic features, solitary HCC with maximum diameter less than 5 cm had the best outcome with the 1st-year survival rate of 100% and the 2nd-year survival rate of 71.4%, while solitary HCC with maximum diameter over 5 cm and multiple HCC had the 1st-year survival rates of 75% and 63.7%, respectively, and the 2nd-year survival rates of 33.3% and 44.4%, respectively. Only one patient was complicated with abscess formation and was cured with antibiotic therapy. No mortality resulted from the procedures performed. CONCLUSION: TAE with ethanol-lipiodol mixture is an economic, safe and feasible method for treating HCC, especially for the patients with smaller solitary HCC or with liver function status of Child-Pugh class A. PMID- 15884126 TI - Association of the myeloperoxidase -468G-->A polymorphism with gastric inflammation and duodenal ulcer risk. AB - AIM: To elucidate the relations between the myeloperoxidase (-468)G-->A polymorphism and the development of duodenal ulcer (DU), and to investigate the impacts of this host genetic polymorphism on the histopathological features of Helicobacter pylori (H pylori)-related gastritis. METHODS: In a case-control study of 115 consecutive DU patients and 182 controls, the myeloperoxidase ( 468)G-->A polymorphism was genotyped. Additionally, gastric mucosal changes were examined according to the updated Sydney System. RESULTS: The two study groups differed in the distributions of myeloperoxidase genotypes (P = 0.008). All six individuals carrying myeloperoxidase A/A genotypes were in the DU group. The carriage of myeloperoxidase allele A and H pylori infection were associated with an increased risk of DU with odds ratios (OR) of 2.3 and 5.8, respectively. The combined risk of the carriage of myeloperoxidase allele A and H pylori infection for DU was 8.7 (95% CI, 3.5-21.8). In the H pylori-infected individuals, allele A carriers displayed higher bacterial density scores (P = 0.04) in the antrum than did non-carriers. CONCLUSION: This work verifies for the first time the association of myeloperoxidase (-468)G-->A polymorphism with antral H pylori density and DU disease. The mechanisms underlying this genetic polymorphism in developing DU disease merit further investigations. PMID- 15884127 TI - Effect of water-soluble contrast in colorectal surgery: a prospective randomized trial. AB - AIM: Postoperative gastrointestinal-tract motility is normally delayed. Early feeding after colorectal surgery has been reported recently, but late feeding is common. Gastrografin not only enhances bowel peristalsis, but also decreases bowel-wall edema. Whether contrast medium allows early oral feeding and reduces the duration of hospitalization requires clarification. METHODS: Fifty patients underwent elective colorectal surgery in a regional medical center. Patients were prospectively randomized into a Gastrografin group or control group (n = 25 each). Patients in the Gastrografin group began their feeding schedule with 100 mL of 5% dextrose water with 100 mL of Gastrografin on postoperative d 3 and were advanced to a full liquid diet when the contrast reached the colon in 4 h. Patients in the control group began their feeding schedule with 200 mL of 5% dextrose water on postoperative d 3 and were advanced to a full liquid diet after the passage of flatus and stool. Nasogastric tubes were inserted for persistent postoperative vomiting. Fullness, nausea, vomiting, complications, time of anesthesia, time of operation, time of mobilization, time of oral feeding, and duration of hospital stay were recorded and analyzed with Student's t-test. RESULTS: In the Gastrografin group, one patient had aspiration pneumonia and one patient had anastomotic leakage resulting in sepsis and eventual death. This mortality was excluded from the subsequent statistical analysis. In the control group, two patients had wound infections. There was no significant difference between the two groups at the time of anesthesia, time of operation, or time of mobilization. There were significant differences between the two groups in the time of oral feeding (3.3+/-0.3 d in the Gastrografin group vs 4.8+/-0.4 d in the control group; P = odds ratio--, 95%CI [-0.5 to +0.7 d]) and in the length of hospital stay (7.6+/-1.1 d in the Gastrografin group vs 10.2+/-1.3 d in the control group; P = odds ratio--, 95% CI [-1.2 to +1.4 d]). CONCLUSION: Gastrografin not only allowed early oral feeding but also reduced the duration of hospitalization after elective colorectal surgery. PMID- 15884128 TI - How can colorectal neoplasms be treated during colonoscopy? AB - AIM: For many physicians who ordinarily treat patients with colonic diseases, colonoscopy is considered a prime study interest. Developments in colonoscopic equipment and methods have led to a large number of endoscopic diagnoses and treatment for colorectal neoplasms. The purpose of this investigation is to evaluate the efficacy and outcomes of endoscopic treatment for colorectal neoplastic lesions. METHODS: From September 1999 to May 2003, 11 447 consecutive colonoscopic examinations in 9864 patients were gathered; totaling 5502 endoscopic treatments for colorectal neoplasms. Macroscopic characteristics of the neoplasms were classified into protruded (n = 3 953), sessile (n = 1402), lateral spreading tumor (n = 139) and depressed lesions (n = 8). Snare polypectomy was conducted in 3984 lesions, hot forcep removal in 1368 lesions, and endoscopic mucosal resection in 150 lesions. RESULTS: Histological diagnoses were 4596 neoplastic lesions (4 376 adenomas and 220 adenocarcinomas) and 906 non neoplastic lesions (891 hyperplastic and 15 inflammatory polyps). For the adenocarcinoma group, 31 instances involved submucosal invasion or resection margin, who received further operations, while 13 surgical specimens discovered no residual tumors. Three perforations and 96 bleedings were found following endoscopic treatment. No procedure-related mortality was found and no recurrent malignancy was found after 14-56 mo follow-up. CONCLUSION: To lower the incidence and mortality of colorectal cancer, endoscopic treatment for colorectal neoplasms is a simple and safe procedure. PMID- 15884129 TI - Monitoring microarray-based gene expression profile changes in hepatocellular carcinoma. AB - AIM: To find out key genes responsible for hepatocarcinogenesis and to further understand the underlying molecular mechanism through investigating the differential gene expression between human normal liver tissue and hepatocellular carcinoma (HCC). METHODS: DNA microarray was prepared by spotting PCR products of 1 000 human genes including 445 novel genes, 540 known genes as well as 12 positive (housekeeping) and 3 negative controls (plant gene) onto treated glass slides. cDNA probes were prepared by labeling normal liver tissue mRNA and cancer liver tissue mRNA with Cy3-dUTP and Cy5-dUTP separately through reverse transcription. The arrays were hybridized against the cDNA probe and the fluorescent signals were scanned. The data obtained from repeated experiments were analyzed. RESULTS: Among the 20 couple samples investigated (from cancerous liver tissue and normal liver tissue), 38 genes including 21 novel genes and 17 known genes exhibited different expressions. CONCLUSION: cDNA microarray technique is powerful to identify candidate target genes that may play important roles in human carcinogenesis. Further analysis of the obtained genes is helpful to understand the molecular changes in HCC progression and ultimately may lead to the identification of new targets for HCC diagnosis and intervention. PMID- 15884130 TI - Variations of very low-density lipoprotein receptor subtype expression in gastrointestinal adenocarcinoma cells with various differentiations. AB - AIM: This study is aimed at investigating the expression and possible significances of very low-density lipoprotein receptor (VLDLR) subtypes in gastroenteric adenocarcinoma tissues and cells with various differentiations. METHODS: Thirty-one cases of gastroenteric carcinoma/adjacent normal tissues were enrolled in the study, which were diagnosed and classified by the clinicopathological diagnosis. The expression of VLDLR subtypes was detected in gastroenteric carcinoma/adjacent normal tissues and three various differentiated human gastric adenocarcinoma cell lines (MKN28, SGC7901 and MKN45) by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. RESULTS: Two VLDLR subtypes, namely, type II VLDLR and type I VLDLR, were found to express changes in gastroenteric carcinoma tissues, their adjacent normal tissue, and gastric adenocarcinoma cell lines as well. Type II VLDLR is predominantly expressed in poorly- or moderately-differentiated gastroenteric carcinoma tissues and gastric adenocarcinoma cell lines, whereas type I VLDLR is mainly detected in well-differentiated intestinal carcinoma tissues and gastric adenocarcinoma cells compared with the adjacent normal tissues. CONCLUSION: The results suggested that the variations of the VLDLR subtype expression might be correlated with the progress and differentiation of gastroenteric carcinoma. PMID- 15884131 TI - Sulindac induces apoptosis and protects against colon carcinoma in mice. AB - AIM: To study the effect of sulindac on colon cancer induction in mice. METHODS: The chemo-preventive action of 80 ppm sulindac fed during initiation and post initiation and 100 ppm sulindac fed during progressive stages of induction of colon carcinogenesis in mice was investigated using 1,2-dimethylhydrazine (DMH). Using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique and PCNA immunohistochemical staining, we observed the apoptotic and proliferative cell density changes at different carcinogenic stages and the effect of sulindac on these two phenomena. RESULTS: Dietary sulindac significantly inhibited the incidence of colonic neoplasmas in mice. Compared with the control group, feeding sulindac during initiation and post-initiation stages inhibited the incidence by 46.7-50.4%, and feeding sulindac during progressive stages inhibited the incidence by 41.1%. Animals that were fed sulindac showed less serious pathological changes than those that were fed the control diet (P<0.01, H = 33.35). There was no difference in the density of proliferating cells among those groups which were or were not fed sulindac. In the same period, feeding sulindac resulted in a higher density of apoptotic cells than feeding control diet. CONCLUSION: Sulindac has an anti-carcinogenic function in mice. Its effect on preventing colon carcinogenesis is better than its effect on treating established tumors. By inducing apoptosis, sulindac inhibited the development of colon cancer and delayed canceration. Sulindac has no effect on proliferation. The anti-carcinogenic properties of sulindac are most effective in the moderate and severe stages of dysplasia and canceration. PMID- 15884132 TI - Detection of T lymphocyte subsets of children with Helicobacter pylori infection. AB - AIM: To study the transformation of T lymphocyte subsets in children with Heliobacter pylori (H pylori) infection. METHODS: The H pylori infection status were determined by a combination of ELISA and Western blot (immunoblot) technique in 98 children and T lymphocyte subsets in peripheral blood were determined by flow cytometrical analysis. RESULTS: There were 75 children positive with H pylori infection and 23 negative in 98 children. Comparing the proportion of peripheral blood T lymphocytic subsets in children with H pylori infection and without H pylori infection, it was found that a higher proportion of CD4 T-cells in infected children (39.02+/-7.71 vs 34.25+/-10.73, t = 2.246,P<0.05) and higher value of CD4 to CD8 T-cells ratio (1.51+/-0.52 vs 1.25, t = 2.104, P<0.05) were present, but there were not significant differences in CD3 T-cells and CD8 T cells (73.11+/-10.02 vs 69.49+/-17.08, 27.22+/-6.07 vs 28.27+/-8.67, P>0.05). CONCLUSION: Th1 cell-mediated immune responses may be induced by H pylori infection in children. PMID- 15884133 TI - Gastric mucosal injury due to hemorrhagic reperfusion and efficacy of Salvia miltiorrhizae extract F and cimetidine. AB - AIM: To observe the gastric mucosal injury caused by hemorrhagic shock and reperfusion and to compare the effect between Salvia miltiorrhizae extract F (SEF) and cimetidine (CI) on it. METHODS: A model of hemorrhage/reperfusion injury was produced by Itoh method. Wistar rats were randomly divided into three groups: 0.9% sodium chloride treatment group (NS group), SEF treatment group (SEF group), and CI treatment group (CI group). Saline, SEF and CI were injected respectively. The index of gastric mucosal lesions (IGML) was expressed as the percentage of lesion area in the gastric mucosa. The degree of gastric mucosal lesions was categorized into grades 0, 1, 2, 3. Atom absorption method was used to measure the intracellular calcium content. Radioimmunoassay was used to measure the concentrations of prostaglandins. RESULTS: IGML (%) and grade 3 (%) were 23.18+/-6.82, 58.44+/-9.07 in NS group, 4.42+/-1.39, 20.32+/-6.95 in SEF group and 3.74+/-1.56, 23.12+/-5.09 in CI group, and the above parameters in SEF group and CI group decreased significantly (IGML: SEF vs NS, t = 6.712, P = 0.000<0.01; CI vs NS, t = 6.943, P = 0.000<0.01; grade 3: SEF vs NS, t = 8.386, P = 0.000; CI vs NS, t = 8.411, P = 0.000), but the grade 0 and grade 1 damage in SEF group (22.05+/-5.96, 34.12+/-8.12) and CI group (18.54+/-4.82, 30.15+/-7.12) were markedly higher than those in NS group (3.01+/-1.01, 8.35+/-1.95; grade 0: SEF vs NS, t = 8.434, P = 0.000<0.01; CI vs NS, t = 7.950, P = 0.000<0.01; grade 1: SEF vs NS, t = 8.422, P = 0.000<0.01; CI vs NS, t = 8.448, P = 0.000<0.01). The intracellular calcium content (microg/mg) in SEF group (0.104+/-0.015) and CI group (0.102+/-0.010) was markedly lower than that in NS group (0.131+/-0.019, SEF vs NS, t = 2.463, P = 0.038<0.05; CI vs NS, t = 3.056, P = 0.017<0.05). The levels (pg/mg) of PGE(2), 6-keto-PGF(1alpha) and 6-keto-PGF(1alpha)/TXB(2) were 540+/-183, 714+/-124, 17.38+/-5.93 in NS group and 581+/-168, 737+/-102, 19.04+/ 8.03 in CI group, 760+/-192, 1 248+/-158, 33.42+/-9.24 in SEF group, and the above parameters in SEF group markedly raised (PGE(2): SEF vs NS, t = 2.282, P = 0.046<0.05; SEF vs CI, t = 2.265, P = 0.047<0.05; 6-keto-PGF(1alpha): SEF vs NS, t = 6.583, P = 0.000<0.000; SEF vs CI, t = 6.708, P = 0.000<0.01; 6-keto PGF(1alpha)/TXB(2): SEF vs NS, t = 3.963, P = 0.003<0.001; SEF vs CI, t = 3.243, P = 0.009<0.01), whereas TXB(2) level in SEF group (45.37+/-7.54) was obviously lower than that in NS group (58.28+/-6.74, t = 3.086, P = 0.014<0.05) and CI group (54.32+/-6.89, t = 2.265, P = 0.047<0.05). No significant difference was shown between NS group and CI group (PGE(2): t = 0.414, P = 0.688>0.05; 6-keto PGF(1alpha): t = 0.310, P = 0.763>0.05; TXB(2): t = 1.099, P = 0.298>0.05; 6-keto PGF(1alpha)/TXB(2): t = 0.372, P = 0.718>0.05). CONCLUSION: Both SEF and CI could inhibit reperfusion-induced injury in gastric mucosa, but with different mechanisms. SEF could not only enhance the protective effect of gastric mucosa, but also abate the injury factors, while CI can only abate the injury factors. PMID- 15884134 TI - Effects of bile reflux on gastric mucosal lesions in patients with dyspepsia or chronic gastritis. AB - AIM: To investigate the influences of bile reflux on profiles of gastric mucosal lesions in patients with dyspepsia or chronic gastritis. METHODS: A total of 49 patients diagnosed with dyspepsia and chronic gastritis underwent 24-h ambulatory and simultaneous monitoring of intragastric bilirubin absorbance and pH values, and then they were divided into bile reflux positive group and bile reflux negative group. Severity of pathological changes in gastric mucosa including active inflammation, chronic inflammation, intestinal metaplasia, atrophy and dysplasia as well as Helicobacter pylori (H pylori) infection at the corpus, incisura and antrum were determined respectively according to update Sydney system criteria. The profiles of gastric mucosal lesions in the two groups were compared, and correlations between time-percentage of gastric bilirubin absorbance >0.14 and severity of gastric mucosal lesions as well as time percentage of gastric pH >4 were analyzed respectively. RESULTS: Thirty-eight patients (21 men and 17 women, mean age 44.2 years, range 25-61 years) were found existing with bile reflux (gastric bilirubin absorbance >0.14) and 11 patients (7 men and 4 women, mean age 46.2 years, range 29-54 years) were bile reflux negative. In dyspepsia patients with bile reflux, the mucosal lesions such as active inflammation, chronic inflammation, intestinal metaplasia, atrophy or H pylori infection in the whole stomach, especially in the corpus and incisura, were significantly more severe than those in dyspepsia patients without bile reflux. Moreover, the bile reflux time was well correlated with the severity of pathological changes of gastric mucosa as well as H pylori colonization in the near-end stomach, especially in the corpus region. No relevance was found between the time of bile reflux and pH >4 in gastric cavity. CONCLUSION: Bile reflux contributes a lot to mucosal lesions in the whole stomach, may facilitate H pylori colonization in the corpus region, and has no influence on acid-exposing status of gastric mucosa in patients with dyspepsia or chronic gastritis. PMID- 15884135 TI - Dissociation, culture and morphologic changes of interstitial cells of Cajal in vitro. AB - AIM: To study the method of dissociation, culture and investigate its morphologic changes in vitro of interstitial cells of Cajal (ICC). METHODS: Enzymatic digestion and Ficoll density centrifugation were used to dissociate ICC from the ileal segment of mice. Factors including contamination, Ca(2+), Mg(2+) and collagenase, and stem cell factor, etc., were investigated. ACK2, the antibody of c-kit, was used to identify the cultured ICC. Both light microscope and fluorescence microscope were used to observe the changes of ICC in vitro. RESULTS: The method for dissociation and culture of ICC in vitro was successfully established. After 24 h, cultured ICC exhibited a few axis-cylinders, and longer axis-cylinders were observed to form synapse of each other after 3 d. More widespread connections formed within 7 d in vitro. The changes of its morphologic character were obvious within 7 d; however, there were no obvious morphologic changes after 30 d. CONCLUSION: Many factors can influence the dissociation and culture of ICC. PMID- 15884136 TI - Analyses of prognostic indices of chronic liver failure caused by hepatitis virus. AB - AIM: To analyze the related indices about the prognoses of chronic liver failure caused by hepatitis virus. METHODS: Retrospectively reviewed 320 cases of chronic liver failure caused by hepatitis viruses. An improved group and an ineffective group (IG) were made to compare and analyze their clinical manifestations, laboratory examination indices and complications. Logistic regression was also carried out. RESULTS: There were significant differences (P<0.05) between the improved group and the IG upon such indices as age, bilirubin, prothrombin time, albumin, alpha fetoprotein, the size of liver and complications (P<0.05). The regression formula was as follows: P = 1/(1+e(-y)) (y = 1.7262 0.0948X(1)+2.9846X(2)+0.6992X(3)+1.6019X(4)+ 2.0398X(5)). (Note: X(1)-Prothrombin activity; X(2)-digestive tract hemorrhage; X(3)-hepatic encephalopathy; X(4) hepatorenal syndrome; X(5)-pulmonary infection.). CONCLUSION: Laboratory examination such as bilirubin, prothrombin time and alpha fetoprotein can be regarded as indices of the prognoses of chronic liver failure caused by hepatitis. Moreover, the regression equation can evaluate prognoses more comprehensively and direct our treatments. PMID- 15884137 TI - Encapsulating peritonitis and familial Mediterranean fever. AB - AIM: To investigate the relationship between encapsulating peritonitis and familial Mediterranean fever (FMF). METHODS: The patient had a history of type 2 diabetes and laparoscopic cholecystectomy was performed one year ago for cholelithiasis. Eleven months after the operation she developed massive ascites. Biochemical evaluation revealed hyperglycemia, mild Fe deficiency anemia, hypoalbuminemia and a CA-125 level of 2 700 IU. Ascitic evaluation showed characteristics of exudation with a cell count of 580/mm(3). Abdominal CT showed omental thickening and massive ascites. At exploratory laparotomy there was generalized thickening of the peritoneum and a laparoscopic clip encapsulated by fibrous tissue was found adherent to the uterus. Biopsies were negative for malignancy and a prophylactic total abdominal hysterectomy and bilateral salpingooophorectomy were performed. RESULTS: The histopathological evaluation was compatible with chronic nonspecific findings and mild mesothelial proliferation and chronic inflammation at the uterine serosa and liver biopsy showed inactive cirrhosis. CONCLUSION: The patient was evaluated as sclerosing encapsulating peritonitis induced by the laparoscopic clip acting as a foreign body. Due to the fact that the patient had FMF the immune response was probably exaggerated. PMID- 15884139 TI - Global assistance. President's AIDS campaign reports success in first year. PMID- 15884138 TI - TT virus infection and pancreatic cancer: relationship or accidental coexistence. AB - TT virus (TTV) was first isolated in 1997 from the patient with acute post transfusion hepatitis. This fact led to the conclusion that the virus was hepatotropic and could be one of the causative agents of acute hepatitis. Afterwards, however, the virus was found in other human tissues and serological studies revealed that it was widespread. Multiple tropisms of TTV and the fact of its high incidence in general population are considered to indicate no medical significance of TTV in human pathology. Here we present a report of two cases of TTV infection in patients who developed pancreas cancer. The patients were hospitalized at the Department of Infectious Diseases due to hepatitis of unknown origin. Since serological and virological markers of common primary and secondary hepatotropic viruses were negative, TTV-DNA was found in serum and was believed to be the only causative agent with probable hepatotropic action. The patients later developed pancreas cancer and they underwent operation. The relationship is difficult to confirm, however the cases we present should be treated as a preliminary report and a comment on the real role of TTV in human pathology. PMID- 15884140 TI - Defamation. Photo alongside teen sexuality article did not defame student. PMID- 15884141 TI - Prevention. Texas approves health bill. PMID- 15884142 TI - Mother stripped of custody for failing to address HIV needs. PMID- 15884143 TI - Securities fraud. Fraud claim over HIV treatment efficacy survives dismissal. PMID- 15884144 TI - Protective order upheld against man for intentional HIV exposure. PMID- 15884145 TI - UNAIDS forecasts future of global progression of HIV/AIDS. PMID- 15884146 TI - Deportation delayed over immigrant's HIV-positive status. PMID- 15884147 TI - Global. Experts offer policy advice to world leaders. PMID- 15884148 TI - Manager cleared of fault over woman's HIV exposure. PMID- 15884149 TI - Bankruptcy. Money used for HIV drugs leaves nondischargeable debt. PMID- 15884150 TI - Social Security. Sedentary job history derails HIV-positive man's SSI claim. PMID- 15884151 TI - Report from the 12th Retrovirus Conference. HIV-associated dementia. PMID- 15884152 TI - Report from the 12th Retrovirus Conference. Perinatal transmission. PMID- 15884153 TI - Invited comment. The "winged" circular anal dilator in stapled hemorrhoidopexy. PMID- 15884154 TI - Ileal resection and small bowel transplantation for large mesenteric desmoid tumor. PMID- 15884155 TI - Sphincter function after surgery for vestibular anus in adults. PMID- 15884156 TI - Taenia solium: characterization of a small heat shock protein (Tsol-sHSP35.6) and its possible relevance to the diagnosis and pathogenesis of neurocysticercosis. AB - A cDNA encoding for a predicted small heat shock protein (sHSP), Tsol-sfISP35.6, has been isolated by antibody screening of a Taenia solium c-DNA library. The clone was a full-length sequence (1172 bp) with an open reading frame of 945 bp and encoded for a 314 amino acid protein with deduced molecular mass of 35.6 kDa, isoelectric point of 5.6 arid the characteristic HSP20/alpha-crystallin domain duplicated. It was highly conserved, with a high sequence similarity with other platyhelminth sHSPs. Western blot analysis, using serum from neurocysticercosis patients (NCC), indicated that the purified Tsol-sHSP35.6 expression product was immunogenic, while in indirect ELISA, using the purified Tsol-sHSP35.6 expression product as antigen and serum samples from pigs and humans, 80% of T. solium infected pigs and 84% of patients with active, or 71% of patients with inactive NCC were sero-positive. The possible relevance of Tsol-sHSP35.6 in the diagnosis and pathogenesis of NCC is discussed. PMID- 15884157 TI - Intradiploic epidermoid cysts of the skull: a report of four cases. AB - Epidermoid cysts are uncommon, benign and slow-growing lesions. They may often reach an enormous size without producing neurological symptoms. Intradiploic epidermoid cysts are nearly 25% of all epidermoids. They are derived from ectodermal remnants that stay within the cranial bones during embryonic development. Intradiploic epidermoid cysts can be located in any part of the skull. Two of our four cases were located in left occipital bone and the others were in the frontal bone. These tumours can occur at any age from the first to the seventh decade of life. Our cases had a mean age of 38.75 years (19-55 years). They may reach great sizes before the initial diagnosis and may produce major neurological signs. For good long-term prognosis, correct radiological assessment and complete removal of the tumour with its capsule are essential. We report four cases of intradiploic epidermoid cysts of the skull and analyse the clinical, radiological features and treatment of these lesions in the light of relevant literature. PMID- 15884158 TI - Perspective. Long, slow learning curve continues on covering the uninsured. PMID- 15884159 TI - Studies show echinacea not beneficial. PMID- 15884160 TI - New prostate cancer treatment preserves function. PMID- 15884161 TI - Increased activity OK for chronic pain. PMID- 15884163 TI - [Joseph Priestley, nitrous oxide and oxygen]. PMID- 15884164 TI - [Tattoos]. PMID- 15884165 TI - In-office fecal occult blood tests fail to detect cancer. PMID- 15884166 TI - Dealing with the "food police". How to keep well-meaning relatives and friends (who worry more about your diabetes than you do) at bay. PMID- 15884167 TI - Constipation: separating fact from fiction. What constitutes the "right" bowel movements? Misconceptions abound. PMID- 15884168 TI - Erectile dysfunction may indicate heart trouble. PMID- 15884169 TI - [About the meeting of the leaders of tuberculosis-controlling services of the subjects of the Russian Federation on progress in 2003]. PMID- 15884170 TI - Variation of long-lived free radicals responsible for the EPR native signal in bone of aged or diseased human females and ovariectomized adult rats. AB - The purpose of this study was to gain insights into the variations seen in the electron paramagnetic resonance (EPR) spectroscopy of the native signals of teeth and bones used for retrospective dosimetry measurements. We determined that changes occur in the long-lived free radicals responsible for the native signal of cortical bone in aging or diseased human females and aged ovariectomized rats. This was done by measuring the magnitude of the broad (BC) and narrow (NC) components of the native EPR signal of bone following chemical extraction, aging, crushing and thermal annealing. Bone from the upper midshaft of femora of young (17-34 years old, n=5) and elderly (70-92 years old, n=18) females was examined. The results showed that the elderly women had significantly higher BC than the younger women (P<0.01). A similar interpretation was made of the data from an aging female rat osteoporosis model. The results for the NC signals were similar. Finally, dramatic decreases in both NC and BC signals were seen in HIV positive and uncontrolled diabetic (one each) patients indicating the need for studying this signal for a broad spectrum of metabolic disorders. Experiments were performed which strongly indicate that iron liganded with organic molecules is the source of the BC signal. Finally, the accuracy achieved in this study indicates that resolving the dosimetric signal (g=2.0018) should be improved by subtraction of the deconvoluted NC and BC signals from the original spectrum. PMID- 15884172 TI - The angular dependence of an Si energy deposition spectrometer response at several radiation sources. AB - An MDU-Liulin spectrometer based on an Si-diode was mainly used during the last few years with the goal to use them for measurements onboard aircraft. To investigate its ability to obtain such measurements, the detector was tested in some radiation reference fields, like 60Co and other photon beams, neutrons of an AmBe and 252Cf sources and in high-energy radiation fields at CERN. Due to the high geometrical asymmetry of the Si-diode semiconductor, an angular dependence of the response would be expected. This work presents analyses and discusses the results of angular dependence studies obtained at the different radiation sources mentioned. It was found that these angular dependences vary with the type and energy of radiation. The influence of these variations on the use as a dosimeter onboard aircraft is also studied and discussed. PMID- 15884173 TI - Discovery of new etchants for CR-39 detector. AB - After reporting the excellent etching properties of molten Ba(OH)2 8H2O as an etchant, we now report some more new and efficient etchants for CR-39 detector. CR-39 detectors were irradiated with fission fragments and alpha particles with a thin 252Cf source. The irradiated detectors were etched in a number of our newly introduced etching solutions as well as in conventionally used 6 M NaOH at 70 degrees C. The newly prepared etching solutions included NaOH/ethanol and NaOH/1 propanol. Processing conditions were optimized for these etchants. From fission and alpha track diameters, bulk etching velocity (VB), track etching velocity (VT), etching efficiency (eta) and their activation energies were determined and compared with that obtained for 6M NaOH at 70 degrees C. PMID- 15884174 TI - [Abstracts of the Neurology Meeting of the French Speaking Community. Marseille, France, 27-30 April 2005]. PMID- 15884175 TI - [Abstracts of the 86th German Radiology Congress. 4-7 May 2005, Berlin, Germany]. PMID- 15884177 TI - Abstracts of the 2nd Tutzing Antiphospholipid Conference. April 18-21, 2005, Munich, Germany. PMID- 15884176 TI - Mapping the air in public schools. PMID- 15884178 TI - [Abstracts of the XXXIXth Congress of the French College of Vascular Pathology. Paris, France, 15-18 March 2005]. PMID- 15884179 TI - Abstracts of the 75th Congress of the European Atherosclerosis Society. 23-26 April 2005, Prague, Czech Republic. PMID- 15884180 TI - Abstracts of the 7th Workshop on Costs and Assessment in Psychiatry. Venice, Italy, March 18-20, 2005. PMID- 15884181 TI - Abstracts of the Society for Academic Emergency Medicine 2005 Annual Meeting. May 22-25, 2005, New York City, New York, USA. PMID- 15884182 TI - Radiotherapy by particle beams (hadrontherapy) of intracranial tumours: a survey on pathology. AB - A review of the principal contributions of radio-therapy of brain tumours by beam particles is carried out. Neutrons, protons and light ions are considered along with their pros and cons in relation to types and locations of brain tumours. A particular emphasis is given to the pathologic studies of their effects directly o n tumours and on the normal nervous tissue, considering mainly the relevant action mechanisms of the radiation types and the requirements of the clinical therapeutic strategies. For comparison the main features of the pathologic effects of radiotherapy by photons are described. From the review it emerges that the new modality of radiation by protons and light ions, because of their peculiar physical characteristics, may represent a new way of destroying the tumour and sparing normal nervous tissue, especially when deeply located and irregularly shaped tumours are concerned. More neuropathological studies are needed in order to better understand the potentiality of the new treatment of modalities. PMID- 15884183 TI - Exposures to atmospheric effects in the entertainment industry. AB - Theatrical fogs are commonly used in the entertainment industry to create special atmospheric effects during filming and live productions. We examined exposures to mineral oil-and glycol-based theatrical fogs to determine what fluids and effects were commonly used, to measure the size distributions of the aerosols, and to identify factors associated with personal exposure levels. In nonperformance jobs in a range of production types (television, film, live theater, and concerts),we measured airborne concentrations of inhalable aerosol,aldehydes, and polycyclic aromatic hydrocarbons, and collected observations about the sites and tasks performed. Both mineral oil and glycols were observed in use on about one-half the production days in the study. The most common effect produced was a generalized haze over the entire set. Mean personal inhalable aerosol concentrations were 0.70 mg/m3(range 0.02 to 4.1). The mean proportion of total aerosol mass less than 3.5 microns in aerodynamic diameter was 61%. Exposures were higher when mineral oils, rather than glycols, were used to generate fogs. Higher exposures were also associated with movie and television productions, with using more than one fog machine, with increased time spent in visible fog, and for those employed as "grips." Decreased exposures were associated with increasing room temperature, with increasing distance from fog machines, and for those employed as "sound technicians." Exposures to theatrical fogs are just beginning to be measured. It is important to consider these exposures in light of any health effects observed, since existing occupational exposure limits were developed in other industries where the aerosol composition differs from that of theatrical fogs. PMID- 15884184 TI - Military medicine in the Spanish-American War. PMID- 15884185 TI - Pushing the boundaries of ecosystems. PMID- 15884186 TI - Parallel experience: how art and art theory can inform ethics in human research. AB - Trends in ethical research involving humans emphasise the importance of collaboration, of involving research subjects, alongside the researchers in the construction and implementation of research. This paper will explore parallels derived from another tradition of investigation of the human: art and art theory. An artist's inquiry into the problems of human research will be described, followed by the application of arguments from art theory to research practice. Recently artist Christine Borland has provided examples in which the lack of collaboration in research has caused injustice. Borland's work reflects these ethical dilemmas and questions the procedures and assumptions involved. In most cases the value of subject anonymity is called into question because it reduces the subjects' control over themselves. The application of art theory, which has already considered these problems, helps question and explore the ways in which the subject turned object of artistic or scientific interpretation can maintain some control and dignity. PMID- 15884187 TI - Principlism or narrative ethics: must we choose between them? AB - This paper addresses a current debate in the bioethics community between principlists, who consider that principles are at the heart of moral life, and narrativists, who see communication at its core. Using a case study entitled "The forgetful mourner" to introduce the tensions between each of these positions, I go on to explain the central tenets of both principlism and narrative ethics. Rather than focus on their respective weaknesses, which many theorists do, I emphasize instead, the contribution that each approach can make to understanding moral life and the process of ethical decision making in health care situations. My ultimate aim is to identify the, sometimes overlapping, skills that both principlism and narrative ethics require on the part of health professionals who deploy them. I conclude that a good principlist has narrativist tendencies and a good narrativist is inclined toward principlism. PMID- 15884188 TI - Mortality and medicine: forms of silence and of speech. AB - Silence can be harmful to patients, their loved ones, and doctors within the contexts of illness and bereavement. I draw from my experience with my son's illness and death to discuss five forms of silence--the silence around the experience of critical illness; the silence between life and death; the silence of doctors; the silence of the dead, and the silence of the ill--and of speech that may emerge in response to these silences. PMID- 15884189 TI - Medicine and the humanities--theoretical and methodological issues. AB - Engel's biopsychosocial model, Cassell's promotion of the concept "person" in medical thinking and Pellegrino's and Thomasma's philosophy of medicine are attempts to widen current biomedical theory of disease and to approach medicine as a form of human activity in pursuit of healing. To develop this approach further we would like to propose activity theory as a possible means for understanding the nature of medical practice. By "activity theory" we refer to developments which have evolved from Vygotsky's research on socially mediated mental functions and processes. Analysing medicine as activity enforces the joint consideration of target and subject: who is doing what to whom. This requires the use of historical, linguistic, anthropological, and semiotic tools. Therefore, if we analyse medicine as an activity, humanities are both theoretically and methodologically "inbound" (or internal) to the analysis itself. On the other hand, literature studies or anthropological writings provide material for analysing the various forms of medical practices. PMID- 15884190 TI - Reprogramming autologous skeletal myoblasts to express cardiomyogenic function. Challenges and possible approaches. AB - Cell transplantation therapy is emerging as a promising mode of treatment following myocardial infarction. Of the various cell types that can potentially be used for transplantation, autologous skeletal myoblasts appear particularly attractive, because this would avoid issues of immunogenicity, tumorigenesis, ethics and donor availability. Additionally, skeletal myoblasts display much higher levels of ischemic tolerance and graft survival compared to other cell types. There is some evidence for improvement in heart function with skeletal myoblast transplantation. However, histological analysis revealed that transplanted myoblasts do not transdifferentiate into functional cardiomyocytes in situ. This is evident by the lack of expression of cardiac-specific antigens, and the absence of intercalated disc formation. Instead, there is differentiation into myotubes that are not electromechanically coupled to neighboring cardiomyocytes. This could in turn limit the clinical efficacy of treatment. This review would therefore examine the various challenges faced in attempting to reprogram autologous skeletal myoblast to express cardiomyogenic function, together with the various possible strategies that could be employed to achieve this objective. PMID- 15884191 TI - Quantifying quality of life for economic analysis: time out for time tradeoff. AB - The "Time trade-off" (TTO), is the most widely used method to "quality adjust" life years for "QALYs" in cost utility analysis. In this paper we ask if it is theoretically likely that the TTO is valid for this use. The TTO consists in a trade off between longevity and quality of life. Firstly, we argue that it is impossible to control for all factors that may influence one's willingness to sacrifice lifetime. Secondly, that longevity and quality of life are too closely interrelated for the hypothetical trade off to reveal real preferences. Thirdly, that the TTO handles the value of a life year inconsistently because it simultaneously assumes that it changes (as an outcome measure) and that it doesn't change (as a currency unit). Lastly, we ask whether the difficulties stem from an inherent contradiction in trying to quantify quality of life. The problems theoretical validity an internal consistency, contrast the use of the results as exact measurements. We conclude that cost utility analysis based on TTO cannot be trusted as a tool for setting priorities in health. PMID- 15884192 TI - Philosophy for medical students--why, what, and how. AB - In this paper the author's biographical choices and experiences of both medicine and philosophy are first described. Then, the concept of philosophy is briefly examined with special reference to medical education. Finally, various ways of teaching philosophy to medical students are illustrated in the light of examples from the author's own teaching. It is argued that, whatever the form of teaching, somehow the content must be linked to the everyday medical world of the students. PMID- 15884193 TI - Consensus and consent. PMID- 15884194 TI - [The role of early diagnosis of gestational diabetes mellitus]. AB - The study aimed to assess fetal outcome in gestational diabetic women and necessity of making screening test in these cases. 94 consecutive women with gestational diabetes mellitus (GDM) made up a study group. All women had been taken care of Perinatology and Gynecology Department of Silesian Medical Academy in period 2002-2003. The study group patients were divided into subgroups. The first group (n = 32) young women, in the second group (n = 62) patients over 30 years old. The following parameters were analyzed: obesity, hypertension, glucosuria, family history of diabetes mellitus, past obstetric history (pregnancy complications, abortions, dead infants and infants over 4000 g). Early glucose tolerance screening during first clinic visit could prevent some diabetes related complications in GDM among women with risk factors. PMID- 15884195 TI - [Prophylaxis of nutrition disturbances project "Time for Health"]. AB - More than half of girls and women treat themselves as obese persons. It is connected with the ideal of slimness, predominating in our culture. The consequences of this cult of slimness involve more and more frequent disturbances in nutrition. Problems, which are the basis for obesity, anorexia and bulimia cannot be related merely to nutrition, weight and body size issues. They are indissolubly connected with low self-esteem, difficulties in one's identity as well as with a lack of interpersonal relation satisfactions. The presented project's aim is prophylaxis concerning nutrition disturbances. It is addressed to girls at the age of 14 and 15. According to the project, classes are organized for the 1st and 2nd year students of secondary schools. The program includes eight meetings devoted to the questions which are significant elements of creating positive image of self-image, a feeling of one's identity as well as developing abilities to make independent opinions and decisions. These factors may prevent girls from following trends e.g. connected with the cult of slimness. PMID- 15884196 TI - [Psychological project supporting weight reduction as a significant element in the prevention of secondary obesity]. AB - Psychological factors may contribute in creating and maintaining of overweight that often results of disturbances of individual's psychosocial functioning. That is why psychological impact should be a significant element of both obesity treatment and prevention of recurrence. The article presents a project implemented in the Obesity Treatment Centre "Waga" in Katowice. The project acts as a part of a group, complex treatment process leading to weight reduction. Design, target group and the achieved results are presented as well a psychological aspects helping the process of reduction of body weight. PMID- 15884197 TI - [Pregnancy complicated by HELLP syndrome]. AB - HELLP syndrome belongs to the group of pathological disorders associated with pregnancy-induced hypertension and may accompany preeclampsia. The basic criteria for establishing the diagnosis are as follows: H--for hemolysis, EL--for elevated liver enzymes and LP--for low platelets. In this report the authors present the case of a 32 years old primipara admitted to the Obstetrics Clinic complaining of epigastric pain, nausea and vomiting. Medical history revealed previously diagnosed and treated reflux disease. In the 39th week of gestation epigastric pain manifested again, blood pressure was high (150/100) and on the basis of laboratory parameters HELLP syndrome was diagnosed (GPT 319 U/L, GOT 204 U/L, platelet 80 x 10(9)/L, antithrombin III 63.9%, D-dimer (+++)). With this case report, authors wished to point out the importance of early diagnosis and treatment of this rare, but having high percentage of perinatal mortality syndrome. PMID- 15884198 TI - [The prophylaxis of HIV infection in nursing practice]. AB - The prophylaxis belongs to the most effective and beneficial methods of preventing diseases not only from economic point of view. Educated and knowledgeable society is a result of effective prophylaxis. The analysis of the obtained results can be summarized by the statement that human fear limits rational and professional perception of the issues relating to HIV/AIDS in selected professional groups. The development of information, education and counseling influence the HIV/AIDS epidemics. PMID- 15884199 TI - [Selected risk factors for coronary heart disease in a group of women aged 30-65 in Krakow]. AB - Mortality rates due to cardiovascular diseases (CD) in Poland are almost twice higher than in the European Union countries. The mortality caused by CD has paid the attention on the fact that it increases relatively among women in comparison to men. Epidemiological observations conducted in female groups indicated that the ischaemic heart disease (IHD) appears more frequently after menopause. The aim of the study was to evaluate some IHD risk factors in the group of women aged 30-65 from Krakow. The study included 95 women, without IHD diagnosis, voluntarily registered to the practice of nurses group. The menopause occurrence and selected possible to modify CD risk factors as a concentration of the total cholesterol, LDL cholesterol, fasting glucose, C-reactive protein (CRP) and also blood pressure, body mass index and waist circumference were assessed. The smoking of tobacco was estimated by an objective method (the level of carbon monoxide in breath out--smoke check). The level of depression was evaluated with the use of Beck Inventory Scale. The results were calculating using Statistica Pl 6.0 programme. The investigated group, especially women after natural menopause, had higher CD risk factors such as: hypercholesterolemia, systolic hypertension, overweight, tobacco smoking, CRP and depression. The tobacco smoking appeared more often in the group after menopause because of other reasons and in the group of women with regular menstruation. The rate of depression was also higher among women after menopause because of other reasons. PMID- 15884200 TI - [Infections of the male urogenital tract and the problem of infertility in the couple]. AB - The retrospective analysis of group of 164 men from sterile couples was made. Authors compared spermiograms with bacteriological investigations' results (sperm bacteriology and blots from urethra for present of Chlamydia, Mycoplasma and Ureaplasma). The statistical signification of coincidence of leucospermia and one species bacterial infection and dependence between leucospermia and pathological spermiogram were found. The positive results of sperm bacteriological investigations were found in normospermic patients too. PMID- 15884201 TI - [Health behaviors of teenagers from Bytom's high schools and prevention of circulatory diseases]. AB - Circulatory system diseases are the epidemic of our times. According to epidemiologic statistics concerning Poland, they are the major cause of deaths. Among diseases associated with circulatory system, ischemia results in substantial number of deaths. However, we can decrease the risk of ischemia by changing the lifestyle. Our research tries to examine and judge the lifestyle of teenagers from Bytom's schools as well as prevention of circulatory diseases- mainly ischemia is concerned. We examined 205 students from secondary schools. The obtained results are not satisfactory. Lack of knowledge linked with good health manners or even improper and anti-health manners of young people may have significant impact on the development of circulatory system diseases in adult life. PMID- 15884202 TI - [Influence of health education on the occurrence of risk factors for coronary heart disease]. AB - Despite many prophylactic activities, coronary heart disease (CHD) remains the main health problem in Poland and the most common cause of death among men. Risk factors related to life style are the target for the modification of prophylaxis. The main task of health education is to develop the skills and knowledge affecting the future behavior and habits of healthy lifestyle. The aim of this project was to assess the influence of health education on the occurrence of CHD risk factors among healthy young men. Sixty four men (31 submitted to education and physical training group and 33 to the control group) up to 50 years of age, without recognized cardiovascular diseases were selected for the study. At the beginning the presence of risk factors was recorded in each participant. The education (6 weeks) and training program (25 weeks) was conducted and it was finally evaluated after six months by comparing experimental versus control groups. The background data showed that 95% of men had at least one risk factor, the mean number of risk factors in average participant was 8.8, minimum 2 and maximum 17, there was no difference between the experimental and control groups. The educational program limited amount of risk factors in the experimental group (p<0.0001). Positive changes concerned physical activities (p=2E-09), knowledge (p=0.01) and stress management (p<0.05). PMID- 15884203 TI - [Program for early detection of ovarian cancer for women as prophylaxis provided at a municipal hospital]. AB - Neoplasms are still severe social problem; they are one of the most common causes of death (30-40%), they last for a very long time and the symptoms are characterized by significant intensity. There have been many attempts of introducing an effective screening program for early detection of ovarian cancer. This paper describes one of them, taking place in municipal hospital. To create and introduce a complex prophylactic program considering early diagnosis of ovarian cancer (with use of Doppler transvaginal sonography and serologic markers -CA125); to assess the value of various features (TVS, CA125 levels, family history) for sensitivity and specificity of screening method; to assess the effectiveness of Doppler TVS as a screening method; and to describe the group of participants that is of the best benefit in screening program. From October 1997 to December 2000 the Program for Early Detection of Ovarian Cancer, introduced by Foundation "Godula HOPE", gathered 3775 women aged 46 +/- 9.5. Most of the participants were self-referred, but there was the group of doctor-referred patients. Bimanual pelvic examination and Doppler TVS were performed in all participants; in case of doubtful results of TVS--serum CA 125 was obtained. The form created by author was used to gather the most important information of medical and family history. The most of participating women were: above 45 yr. old, married, on middle education level, non-working or mental worker, living in town. 365 adnexal tumors were detected among examined women. Using our own criteria 228 patients were chosen to laparotomy--16 of operated tumors were histologically confirmed to be malignant. Our sensitivity of PI was 93.75%, specificity--89.62%. The values for RI were respectively: 89.61% and 73.72%. Program of early detection of ovarian cancer using transvaginal sonography can be organized in municipal hospital; the most important feature of the examination (for screening program) is the grayscale sonographic morphology and the Doppler parameters are of additional value. The best cut-off values of PI and RI were 0.8 and 0.5 respectively. The great meaning of family history of ovarian cancer was confirmed. Demographic results suggest that program should be modified to be available for older patients on low level of education with family history of ovarian or breast cancer. PMID- 15884204 TI - [Analysis of some pro-health behaviors of pregnant women after a childbirth education class]. AB - Nowadays the school of childbirth is one of the most important forms of antenatal education. The aim of the study was to compare some pro-health behaviours of women after courses at school of childbirth with pro-health behaviours of women without such education. A group of 129 women after courses at schools of childbirth who were hospitalized in the Chair and Department of Obstetrics and Perinatology of Pomeranian Medical University in Szczecin were introduced into the study. The control group contained 165 women who did not take part in any antenatal education. The questionnaire was a research tool; it was completed 2-5 days after childbirth and also patients' documentation was analyzed. The test results were statistically analyzed. Investigations showed that woman attending classes at school of childbirth were highly aware of pro-health behaviours during pregnancy; they often changed their lifestyle during pregnancy, they smoked less frequently, they had more appointments with their obstetricians and they earlier had their first medical visit. It seems necessary to intensify the classes of nourishing during pregnancy at schools of the childbirth, because no statistically significant differences in the increase of body mass were showed. Moreover, it is necessary to educate every pregnant woman of the hygiene, nourishing and stimulant abuse during pregnancy not only at schools of the childbirth, but also during obstetric consultation. PMID- 15884205 TI - [Sources of women's knowledge relating pregnancy and childbirth]. AB - Future parents have the huge range of possibility to afford theoretical knowledge from different sources: books, periodicals, advice of close, doctor's advice or the midwife in outpatient clinic and more. One of the most professional sources of knowledge preparing to parenthood is the delivery school. The aim of investigations is the analysis of sources of women's knowledge preparing to maternity at schools of childbirth and without such form of education. Investigations were conducted on Maternity Ward at the Chair and Department of Obstetrics and Perinatology of Pomeranian Medical University in Szczecin. Investigations were having taken over 129 women after delivery school. Control group was 165 women, who did not take any antenatal education. Investigations appeared that women participating in activities of the delivery school more often read professional magazines and books, watching films preparing to birth control. This is probably the result of significantly frequent higher education in this group as well as the projection of educational films in delivery school. Moreover woman without preparation in delivery school more often had to need the help of hospital staff during delivery. PMID- 15884206 TI - [Sources of women's education about the puerperium and breast feeding]. AB - Future parents have a wide range of possibilities in gaining theoretical knowledge from books, magazines, relatives' advice, doctor or midwife' consultations in outpatient clinics. The most professional ways of getting skills in that matter are birth schools. The aim of this study was to analyze the sources of knowledge about puerperium and breast feeding at birth schools in comparison to mothers who did not attend such training. The study was undertaken in the Department of Obstetric and Perinatology in Szczecin. Group I (investigated) consisted of 129 women who finished birth school training course and group II (control) consisted of 165 women with no formalized prenatal education. A questionnaire and medical histories were used in our analysis. The results showed that the birth school attendants were more likely to use a lot of education sources in the field of breast feeding and that some law aspects in puerperium and puerperium pathology need more attention because books and magazines are the main source of knowledge in that matter instead of birth schools classes. PMID- 15884207 TI - [Smoking among secondary school students in Stalowa Wola]. AB - Smoking is recognized as major, the most prevalent, and at the same time the only removable factor of many diseases. The major causes of starting smoking are: stress, curiosity and staying among people who smoke tobacco. The research was conducted among students of secondary school in Stalowa Wola in the form of an anonymous poll. The sample group included 2763 people. The group of smokers included 334 people (12.09%). Results of the research were subjected to a precise statistical analysis. The fact that 24.55% young age smokers smoke more than 20 cigarettes a day is worth stressing. In spite of short time of tobacco smoking, the investigated group has reported some negative health ailments. An attempt was made to identify the combination of features affecting smoking, including sex, origin, company of smokers, economic conditions (p < 0.05). The statistical analysis revealed a significant correlation between the number of cigarette smoked, economic conditions and the intention to stop smoking (p < 0.05). The obtained results may be helpful in preparing effective educational and preventive project among students, which can eliminate or lower the influence of factors predisposing to start and continue smoking tobacco. PMID- 15884208 TI - [Women's knowledge of health behaviors in the puerperium]. AB - The study was conducted on one hundred women hospitalized in obstetrical departments of hospitals in Bialystok. Material was collected using anonymous inquiry in the accordance with methodology of questionnaire creation. Over half responders (55%) correctly defined puerperium, however 19% of them couldn't explain what it means. Near 1/3 women answered that bloody puerperal excrements may be present to the end of puerperal period, 41% of women answered that during puerperium they are allowed to bath and only 22% of them considered physical exercises to be profitable. 52% of responders knew that newborn child should be breastfed for six months and that breastfeeding prevents breast from incidents of occlusion. 20% of mothers considered breastfeeding to be preventive from defects of bite and dental caries. CONCLUSIONS: 1. Definition of puerperium caused problems for most of women. 2. The period of puerperium affected the mode of lifestyle and hygiene of women but most of them did not consider physical exercises to be profitable. 4. The most of women knew that breastfeeding is necessary for appropriate growth of their children. PMID- 15884209 TI - [Phytoestrogens--an alternative to hormonal replacement therapy]. AB - Perimenopausal period is associated with the reduction of endogenous estrogens which might lead to many disorders of general health in women. Traditional hormone replacement therapy (HRT) is effective for controlling vasomotor symptoms and reducing the risk of cardiovascular disease and osteoporosis in postmenopausal women. However, according to the latest studies, many women are reluctant to initiate this therapy because of concerns regarding the benefits and risks considering contraindications and side effects of it. Therefore, a lot of studies were carried out to find the influence of phytoestrogens on menopausal symptoms. Phytoestrogens are defined as naturally occurring compounds, found in plants; they have a variety of activities: estrogenic and antiestrogenic. Could phytoestrogens be used as an alternative to hormonal therapies for the management of menopausal symptoms? PMID- 15884210 TI - [Effect of the body weight of pregnant women on delivery and the birth state of the newborn]. AB - Systematic and lately even rapid growth of the number of overweight people has been recently observed. In the economically well-developed countries fatness is the real problem of the contemporary man. The evaluation of the body weight of the pregnant women and its influence on the delivery and the birth state of the infants has been the aim of the study. 139 cases of pregnant women with physiological pregnancy have been analyzed. The pregnant women were divided into 4 groups according to the body weight at the early pregnancy which according to WHO criteria was expressed by the body mass index (BMI). The body mass of the delivering woman, date of delivery, way of delivering and probable delivering complications, performed obstetrical procedures, estimated blood loss of the delivering woman, sex, weight and length and the birth state of the infant according to the Apgar scale were evaluated prospectively. Analyzing the body mass of the women at the early pregnancy, underweight was stated at 55 women (39.57%), the proper body weight at 66 cases (47.48%), overweight at 12 cases (8.63%) and the fatness at 6 cases (4.32%). The average body mass index (BMI) at the early pregnancy was 21.63 kg/m2 and its increase during pregnancy was 5.95 kg/m2. It has been found out that the increase of body mass during pregnancy corresponds to the body weight before pregnancy and amounts to 16.22 kg on average. The study has shown more cases of surgical deliveries, greater intrapartum blood loss in the group of overweight and fat women, and greater harm to the vulva/vagina during delivery in the group of underweight women. The best delivery state of the babies has been found out in the group of women with the proper body weight. PMID- 15884211 TI - [Breast cancer prophylaxis among nurses]. AB - Breast cancer is one of the most frequent malignant diseases among women. Breast cancer detection at an early stage is possible thanks to the active attitude of women themselves. The breast self-examination is a cheap and easy method for early detection of breast tumors. At the same time it can protect women from the development and long-term complications of that disease. The aim of the study was to evaluate the nurses' knowledge of the breast cancer prophylaxis and a range of its use for themselves. The research was conducted among 193 nurses taking part in qualification courses with the use of a questionnaire consisting of 20 questions. The results showed that 63% of nurses knew almost everything about the early breast cancer detection. Almost 50% of nurses did self-control regularly but unfortunately, not all of them did it correctly. Only 38% of participants were examined in outpatient clinics. 33% had mammography and 41% breast's USG. The results of the study are not optimistic. Despite medical education, the nurses are not knowledgeable enough about prevention and breast cancer detection. It also appeared that the nurses do not use their practical knowledge. PMID- 15884212 TI - [Decreasing trend of cardiovascular mortality in Poland in the years 1980-2001]. AB - In Poland, mortality due to cardiovascular diseases (CVD) has been systematically decreasing since 1990. The apparent change has been subject to a number of investigations during the last decade. The potential explanatory factors include favorable effects of socio-political and economic changes in Poland, decrease in the occurrence of risk factors for cardiovascular diseases, modification of diet and lifestyle, as well as advances in the treatment of cardiovascular diseases, particularly in their acute, life-threatening stages. Hypothesis regarding a beneficial evolution of the occurrence of risk factors is not universally accepted. The lack of convincing explanation of the trend justified a revision of the mortality pattern, including analysis of the specific components of the overall category of CVD. The findings showed that during the period in question (1990-2001) some important specific mortality rates have increased and not decreased. The change was particularly apparent in terms of ischaemic heart disease and stroke. Assessment of available data reveals also a decrease in mortality rates due to hypertension and due to diseases of arteries. The results of analysis suggest a caution in exploring and interpreting of overall trend in cardiovascular mortality in Poland, during the last decade. Beneficial changes appear to be less promising when the mortality profile is examined via the contribution of specific diseases, components of the overall category of CVD, especially ischeamic heart disease and the stroke. The findings justify further exploration, involving also a potential effect of diagnostic shift in classification of causes of death. PMID- 15884213 TI - [Thermography in the early detection of breast cancer--our own experiences]. AB - In combination with X-ray mammography and clinical examination thermography adds to the accuracy of breast cancer diagnosis. Thermographic examinations can be repeated at short intervals with no radiation hazard to the patient. However, performing and interpreting thermograms requires meticulous training. It was found that women with an abnormal thermogram are at a higher risk and have a poorer prognosis. A change in the thermal pattern is often the earliest sign of a cancer. The authors believe that the fault lies in misinterpretation of the thermogram, rather than the thermogram itself. Computer stimulations could be an adjunct tool to help the clinician in the interpretation. PMID- 15884214 TI - [Assessment of social and professional support offered to patients during their stay in a rooming-in at a maternity ward]. AB - The improvement of quality of care on maternity wards concerns not only the medical activities. Professional activities minimalizing the occurrence of difficult situations of woman in puerperal period are of great importance as well. The aims of the study were: 1) to evaluate the accessible social support and the need of professional support in women during their staying on maternity ward in rooming-in system; 2) to establish the correlation between accessible social support the need of professional one. Studies were conducted at one of maternity wards in Szczecin working in rooming-in system. Studied group were primiparas (n = 100), reference group consisted of multiparous females (n = 100). The following tools were introduced into the study: Modified Social Support Scale and Author's questionnaire on Demand for Support in maternity ward in rooming-in system. The results of investigations show, that the need of professional support was more often expressed in the study group (p < 0.001). It reached an average- high level. There was no significant difference concerning an accessible social support in both groups. No correlation was showed between accessible social and professional support. CONCLUSIONS: 1. Primiparas require special puerperal care and professional support as well. 2. High, accessible social support has no influence on need of professional support in women during their stay on maternity ward in rooming-in system. PMID- 15884215 TI - [Secondary prevention of myocardial infraction]. AB - Results of numerous epidemiological research prove that the development of civilization, density of population, constant changes of human diet based on increased consumption of processed, high-energy food rich in animal protein, sugar, saturated animal fat and change of life standard that is characterized by limiting physical activity and energy output and widespread pollution became favorable conditions (if not causative) increasing the rate circulatory system morbidity, including ischemic heart disease and myocardium infraction. The purpose of presented research is an attempt to answer a question how the patients after myocardium infraction self-estimate their previous style of life. The paper also tries to find the relation between family record of sickness with dietary preferences and physical activity. In purpose to collect the empiric material the scientists used a method of diagnostic soundings. The author's questionnaire was used as a technique of the survey that was performed on a group of 100 patients after myocardium infraction. The patients were chosen randomly and the survey was accomplished in hospitals of Slupsk and Koszalin. Taking care of proper health condition by patients after myocardium infraction is highly unsatisfactory. The results show that in spite of myocardium infraction, 30% of respondents still smoke, 30% use alcohol and 40% use inappropriate diet. Dietary references of the respondents prove the necessity of providing more education concerning healthy diet in everyday life. The respondents ceased systematic physical exercises and the majority of them (62%) does not exercise regularly (32% cured by preservative method and 30% invasion method). PMID- 15884216 TI - [Health risk behavior among students of vocational schools]. AB - Behaviour of an individual person has a significant influence on the health. The stimulant abuse and lack of various safety regulations have an inefficient influence. The aim of the study was to identify the frequency of health risk behaviour among adolescents as well as the assessment of their level of health awareness. The investigated population consisted of third grade pupils from vocational schools in Poznan and Wagrowiec. Stereotypically, vocational schools' pupils are evaluated to have a low level of health awareness and to behave in unhealthy way. It was intended to find out if city versus small town young population differs on account of their health awareness and behaviour. A questionnaire was the basic tool to collect data. It was chosen because of its anonymity in order to guarantee honest answers. The results were analysed statistically using SPSS and Statistica programmes. Chi square and significance tests between two differences were used. The differences were statistically significant when p < 0.05. The results were subsequently analysed according to following independent variables: place of living, sex, parental education, the family economic self-evaluation. CONCLUSIONS: 1. The level of health awareness among the investigated population can be generally described as medium. 2. There were no differences in the level of health awareness between the city youth and the small town youth. 3. A large proportion of the youth had unhealthy behaviour. 4. City dwellers tended to behave recklessly more often. 5. The high level of health awareness was connected to less frequency of health risk behaviours. PMID- 15884217 TI - [Herbich cervical pessary--method of therapy for cervical incompetence and prophylaxis of prematurity]. AB - Cervical incompetence is often complication in the II and III trimester of pregnancy and is a cause of 16-20% of premature deliveries. Treatment of incompetent cervix contains surgical methods ( cerclage and others) and conservative management as follow: tocolytics, hormones, bed rest and mechanical factors (pessary, rings, balloons etc). The aim of the study was evaluation of therapeutical efficacy Herbich cervical pessary in the cases of cervical incompetence I and II degree in II and III trimester of pregnancy. Study included 73 pregnant women with cervical incompetence diagnosed on the basis of clinical examination and measurements of cervical length during transvaginal ultrasonography (Voluson 730 Kretz, ATL HDI 1000). After diagnosis we inserted into vagina pessary size 1 or 2 around incompetent cervix. We divided examination group into primipara and multipara. In our research we obtained no statistically significant differences between both groups in the percentage of premature deliveries, the mode of delivery and prolong time of pregnancy. High percentage of prematurity (35.7%) was a cause of two fetal death (2.75%) delivered in the 25 week of pregnancy. Cervical pessary prolonged time of pregnancy about 12-17 weeks and for this reason almost all labors were after 33 weeks of pregnancy (89%). Cervical pessary of Herbich is a safe and effective method of treatment for cervical incompetence of I and II degree. This pessary decreases percentage of prematurity in the high risk group of pregnant women with incompetent cervix. PMID- 15884218 TI - [Standard of living as one of the risk factors for breast cancer]. AB - The aim of the study was to estimate the level of knowledge about the influence of standard of living, and particularly a type of nourishment obesity, taking drugs, on risk of the breast cancer. The survey was conducted among 186 women, different in relation to their age, state, permanent residence and education in midwife profession. The questionnaire was the main research tool in this survey. The knowledge of risk factors (obesity increasing a breast cancer spread) depends statistically on the age, state and the education level of the examined women (p < 0.05). The knowledge of risk factors (the type of nourishment and taking drugs increasing breast cancer spread) depends statistically only on the level of education of the examined women (p < 0.05). PMID- 15884219 TI - [Knowledge of breast cancer risk factors as one of the conditions in undertaking prophylactic treatments among midwives]. AB - The aim of the study was to estimate the knowledge of breast cancer risk factors among midwives on different education level. A diagnostic survey was undertaken with the use of questionnaire technique among 186 persons representing different education level of midwife profession--licentiate students (37.63%), master's degree students (29.03%) and participants of family nursing course (33.33%). The collected data were submitted to statistic analysis and chi2 test was used to check the significance of investigated features. The general knowledge about breast cancer risk factors among persons representing different education level of midwife profession is satisfactory. The stage of education correlates with the level of knowledge about risk factors connected with family transmission, the age influence, menopause time and breast self-examination in the investigated group (p < 0.05). PMID- 15884220 TI - [Fetal pulmonary artery blood flow depending on fetal lung maturity]. AB - The ultrasonographic assessment of fetal lung maturity by evaluating the elasticity of lung tissue, Dynamic Lung Score (DLS) has been being performed since 1986 in ObGyn Department in Tychy (Medical University of Silesia). The lung elasticity is evaluated on the cross sections of fetal thorax, on the level of heart ventricles. The result of the evaluation is given as the three degree scale, in which I degree indicates the lack of elasticity and fetal lung immaturity, II degrees indicates partially expressed elasticity, corresponding with incomplete maturity of lung tissue, and III degrees represents full elasticity and indicates complete maturity of fetal lungs. This study was designed to compare fetal pulmonary artery blood flow with the maturity of fetal lung tissue evaluated during the ultrasonographic assessment of fetal lung tissue elasticity. The examination was performed on pregnant women, beginning on 27th week gestation. During the examination the mean Pulsatility Index was decreased, particularly in fetuses with II degrees lung maturity. The Resistance Index (RI) was found to be stable and independent of gestational age. In the group with I degree lung maturity (DLS I), the mean PI = 2.643 (+/- 0.229), mean RI = 0.879 (+/- 0.036), in DLS II group PI = 2.039 (+/- 0.262), RI = 0.868 (+/- 0.037), and in DLS III group PI = 2,500 (+/- 0.100), RI = 0.900 (+/- 0.100). Comparing the ultrasonographic evaluation of fetal lung maturity with fetal pulmonary artery blood flow allows more accurate assessment of fetal lung maturity. Fetal lung maturity can not be evaluated univocally on the basis of blood flow assessment. Because of the divergence of blood flow parameters further studies including bigger population seem to be necessary for verification of the results and for establishing the reference values. PMID- 15884221 TI - [Prophylaxis of Internet sex addiction]. AB - Sexual behaviour, sexual relationships in Internet are regulated by specific rules. The aim of this study was to analyze the problematic Internet-related sexual behaviours, the evolution mechanisms of sexual behavior; it describes the character of an addicted person and consequences of these changes. The development of Internet, high sex-technology are very dangerous for many young people. It seems necessary to educate and promote reliable knowledge about sexuality during school education and the whole life. PMID- 15884222 TI - [Prevention of premature birth in the population of pregnant women in Silesian region]. AB - A great number of premature deliveries is one of the main problems in present-day perinatology. Although the ability of premature babies to survive has increased, they are vulnerable to many complications. Statistical data showed that the grade of premature deliveries and related complications in Silesian region is one of the highest in our country. Therefore, it is necessary to offer the intensive and systematic health care for pregnant women and establish the prophylactic programme for premature deliveries. The aim of this article is to evaluate the level of knowledge, skills and motivation to follow the prophylaxis directions among pregnant women. PMID- 15884223 TI - [Prevention of intrafamilial childhood sexual abuse]. AB - At work we took up the matter of sexual harassment of children in the family. We presented the history of incest contacts, reasons, conditions causing incest, the perpetrator, his methods and kinds of his actions.We took into consideration description of victims, physical and psychological symptoms of sexual harassment and its effects. We paid attention to effective methods of prevention of incest behavior, diagnostic actions taken in order to confirm any offence and therapy of victims emphasizing role of health service staff. PMID- 15884224 TI - [Prevention of abortion]. AB - At work the subject of abortion has been taken up. We took into account the history of this phenomenon, medical and legal aspects. We showed the reasons for abortion. We analysed abortion: permitted by law and illegal. We paid a lot attention to preventive actions taken in order to prevent women and families from harmful consequences of abortion. PMID- 15884225 TI - [Prevention of intrapartum fetal hypoxia by using electrocardiography]. AB - The biophysical diagnostic methods are an unusual element of evaluation of the fetus state in the time of pregnancy and delivery. Nowadays, the applied methods of fetus supervision during labor don't ensure the permanent analysis and they don't deliver diagnostic information about fetus hypoxia. The purpose of avoiding the risk of fetus injury and its hypoxia should be developed by technologies which allow making a diagnosis of fetus condition and to detection of intralabor hypoxia, So, especially all new methods are indicated, which make easier the diagnostic in a permanent mode and which allow for hypoxia expectation and they also allow to reduce the number of unjustified medical interference. One of these methods is fetus electrocardiography. PMID- 15884226 TI - [Prevention of iron deficiency anemia--influence on the course of pregnancy, delivery and the infant's status]. AB - BACKGROUND: Iron deficiency is probably the most common nutritional disturbance in the world. At the highest risk, irrespective of economical status, are women at the reproductive age--especially those who are pregnant. Retrospective analysis of a course of gestation, delivery and the infant's status depending on mothers' iron supplementation during pregnancy. Medical notes of healthy 860 pregnant women hospitalized at term in the Clinic of Perinatology and Gynaecology, Zabrze, Poland were analyzed. Patients were divided into two categories: Group I (n = 610) were those having iron subjunction and Group II (n = 250) those who did not. Mean gestational age at the delivery was significantly lower in those without the prophylaxis. These patients had significantly higher rate of preterm deliveries and caesarean sections. Macroscopic abnormalities of the afterbirth were commoner in this group as were bleeding during III labour stage and indications for postpartum curettage. insignificantly lower mean birth weight and 1st/5th minute Apgar scores had children born to women without supplementation. Routine iron supplementation in women with initially normal haemoglobin levels seems to be a safe strategy for preventing perinatal complications. PMID- 15884227 TI - [Influence of the prevention of maternal periodontitis on the risk of preterm delivery]. AB - The premature labour has been regarded as a highly problematic clinical event not only due to its relation with perinatal mortality but also in the light of the remote results of the prematurity. Evaluation of the influence of active periodontitis on a preterm delivery risk. Retrospective assessment of the preterm delivery risk in a group of pregnant women with clinically confirmed periodontitis and healthy otherwise was performed. Further risk assessment was stratified on a basis whether prophylactic measures against this dental pathology were introduced (n = 30 of women were treated). The control group consisted of 30 pregnant and healthy women. Patients with active periodontitis and having a prophylactic treatment had a 4-fold increase of a preterm delivery risk compared to those without this dental pathology. The risk was 12 times higher in the group with advanced changes and no treatment. According to the results, periodontitis should be regarded as a systemic disorder capable of affecting a pregnancy so prevention procedures should be introduced as soon as the problem is recognized. Such a procedure involving dental assessment in described population could significantly decrease the number of preterm deliveries. PMID- 15884228 TI - "Saba"--application of knowledge and state-of-the-art technologies in the field of psychiatry for development of new diagnostics prevention and therapeutic tools for schizophrenia. AB - The aim of project is to build a European network, which will integrate the research capabilities of a group of research institutes and university departments to provide an infrastructure for the highest quality research in psychiatric disorders, particularly in schizophrenia and schizoaffective disturbances. This network will integrate original computer expert advisory system called "Saba" with modern brain imaging techniques and neurophysiological methods, which allows for the delineation of specific subtypes and particular episodes of mental disorders and their neural bases will be studied by state-of the art (high tech) imaging techniques. This approach will lead to new investigatory, diagnostic and therapeutic techniques. Together the members of this network will comprise an unmatched critical mass of human and other resources aimed at fundamental and applied research into a group of disorders, which impose a huge burden on social and material capital. The relationships and mutual responsibilities between neuroscience and the society it serves will be addressed specifically. Top brain research is performed at several locations in Europe. In particular, in the area of linking classical psychiatric and psychological assessment methods and the newest brain imaging techniques in mental disorders, major progress can only be made when various research groups join their efforts. Large-scale studies using different databases are critically required, which demands standardization of the description of mental disorders and of the applied techniques and methods of analysis. Imaging techniques, including functional MRI (fMRI), Evoked Potentials (EPs), brain mapping, and the computer gathered information will be shared, standardized and further developed within the network. Developing new information technology tools for simulation, visualization and data-mining will be required to enable effective search for links between mental disorders and brain characteristics (function, structure) in very large scale data-sets acquired and stored in various research facilities. PMID- 15884229 TI - The integration of the computer advisory system together with neuroimaging procedures, neurophysiological markers and psychological cognitive tests in order to obtain evidence based system for the physician treating affective disturbance. AB - WHO has classified depression disturbances--due to widespread existence and serious medical consequences and resulting social and economic effects--as a priority health problem in all the developed countries. The significance of the depression disturbances is fully comparable to such illnesses as heart diseases, cancer and HIV infection. The research made in the USA (NIMH programmer) shows that first contact doctors are not able to detect depression in relation to 50 70% of the people ill with it. Also the research made in other countries shows that a doctor properly identifies only one out of four persons with clear indications of depression. The wrong choice of the antidepressant drug may result in inefficient therapy and in growing risk of suicide. In 1993 the analysis of 50 best selling medicines in the EU made by two Italian pharmacologists showed that in France and Italy in over 45% of the cases the medicines with doubtful efficiency are applied. The aim of our project is to build a European network, which will integrate the research capabilities of a group of research institutes and university departments to provide an infrastructure for creation of the computer advisory system for diagnostics of affective disorders. This network will integrate our original computer expert advisory system called "Salomon" with modem brain imaging techniques and neurophysiological methods, which allows for the delineation of specific subtypes and particular episodes of mental disorders and their neural bases will be studied by state-of-the art (high tech) imaging techniques. This approach will lead to new investigatory, diagnostic and therapeutic techniques. PMID- 15884230 TI - [Selected aspects of diagnosis and treatment of transsexualism in Poland]. AB - Transsexualism is a form of sexual identity disorder. In this disorder sexual identification psychological level is inadequate to biological level. One man out of 30000 and one woman out of 100000 ones is transsexual. The etiology of transsexualism has not been known yet. The treatment is complex and a cooperation of the specialist team is necessary. An operation is a second step after the diagnosis of transsexualism (psychological, psychiatric and sexological), hormonal therapy end Real Life Test. Follow-up therapy with the psychologist and supportive therapeutic group is extremely important after the operation. PMID- 15884231 TI - [Prophylaxis of prostate diseases]. AB - According to the WHO, what second man after fortieth year of life, which, suffers on disorder of prostate gland fifth can hear once "my the mister of cancer of prostate". The cancer of prostate gland is this second, after cancer of skin, the most often recognizable tumor at men, and second, after cancer of lungs the most often bringing to death. It is the purpose of article the performance of preventive workings inhibitory the excess of prostate gland, as well as reducing the risk of falling ill on neoplasm disease of prostate gland. PMID- 15884232 TI - [The need for health promotion of medical students and proposal for prevention of addiction]. AB - Alcohol, drug addiction or any other addiction to psychoactive drugs constitutes one of the most serious social problems in Poland. Despite systematic and more professional addiction prophylaxis, increased consumption of psychoactive drugs can be observed among children and youth. Prophylaxis is often limited to giving information on what must not be done and it does not include suggestions connected with constructive ways of coping with problems or of getting satisfaction. The problem of using psychoactive drugs by medical students is often neglected. Only few authors deal with this problem. The aim of the presented project is to educate medical students how to cope with difficult situations, form mutual reliability in a group, better self evaluate and wide the range of learning techniques. PMID- 15884233 TI - [Influence of selected family environment factors on the child's speech development]. AB - All environment strata, natural, cultural and social environment affect the individual in typical way and create his specific reactions as well as mental experiences. A speech, as one of elements enabling adaptation to life in community is formed individually with particular children. The basis of such process is always the possibility of the intercourse with speech. The aim of undertaken subject was the analysis of influence of selected family environment factors on the child's speech development. The study was conducted with children of six and seven year old, where speech should be mastered with fixed pronunciation and proper speech technique. The following research problems have been assumed: 1) how does the speech competences' development with children of six and seven proceed, 2) in which level do the following factors of family environment (parents' age, parents' education, family's financial conditions) influence the child's speech development, 3) does the family's structure influence the proper child's speech development. The following research methods were applied: observation, interview and questionnaire. Applied research tools may be described as follows: inquiry sheet of questionnaire, child's speech card, pictorial test, test chi2. The presented work is a trial of analysis how the selected family environment factors influence the child's speech development. PMID- 15884234 TI - [Assessment of mass-scale primary prevention of nervous system defects]. AB - Nervous system defects are one of the main reasons of neonate mortality in Poland. In reply to that fact a special program for prophylaxis of nervous system defects was established in 1998. It includes three activity fields for women at reproductive age. These are following: propagation of nourishment rich in vitamin B and folic acid, recommendation for the day-to-day taking of 0.4 mg folic acid, improvement of the nutritive food value by adding such nutrients as folic acid. The aim of this article is to evaluate the efficiency of the prophylaxis of the nervous system defects. PMID- 15884235 TI - [Prophylactic role of the specifics and dynamics of health-related behaviors in patients with lower limb arteriosclerosis]. AB - The article presents longitudinal research into peculiarity and dynamics of health behaviours in patients suffering from atheromatosis of he lower limb arteries. The aim of the research was to show how the onset of the disease influenced patients to change their health behaviours. The survey was conducted in 2002-2003 and it included 114 patients of the Outpatient Vascular Clinic at Biziel Hospital in Bydgoszcz and a homogeneous group of 114 healthy people. The analysis shows that the level of health behaviours, considered to be the main cause of the disease, varies according to the health of surveyed people. The direction of changes (improvement in health behaviours) suggests that the onset of the disease is a traumatic event which leads to these changes. The dynamics of changes in health behaviours shows that the highest level of improper health behaviours is usually observed before the onset of the disease and then health behaviours improve. However, the dynamics slows down as the disease progresses. Additionally, patients tend to declare better health behaviours in the future but these declarations are not fulfilled unless patients are active or exposed to stress. The results of the research show that it is necessary to make use of the behaviour changes caused by traumatic event. They also show the need for an interactive prophylactic programme which will prolong the tendency towards changes in pro-health behaviour. PMID- 15884236 TI - [The knowledge of preventive measures and early detection of breast cancer among students of the Medical University in Lublin]. AB - A breast cancer is the most common neoplasm among women. It occurs in nearly 40 out of 100000 cases. A number of breast cancer cases have doubled for 30 years in Poland. According to long-term statistical data, it can be assumed that that number will be increasing. Approximately 7% of women are cured owing to preventive care in western countries. The diagnose at the early stage gives a chance to treat it effectively. Therefore, it is essential to increase the public awareness about the danger of the cancer and possible ways of its early detection. It should be an important task of doctors, midwives and nurses, especially young doctors to be thoroughly prepared in the field of risk factors, preventive care and early detection of breast cancer. PMID- 15884237 TI - [Value of the photodynamic method in the diagnosis of superficial bladder tumors]. AB - Photodynamic diagnosis (PDD) is one of most precise method of neoplasms recognition. The aim of the study was to estimate the value of 5-amino levulinic acid in diagnosis of superficial bladder tumours. Since 1996 to March 2004, 597 cystoscopies were performed with 5-amino levulinic acid in 151 patients with superficial bladder tumours (102 men and 49 women) aged 19-87 years old. 132 pts were treated by PDD after transurethral resection of bladder tumours (TURB). 749 specimens were taken from mucosa during PDD. Before PDD 50 ml of of 5-amino levulinic acid solution was instilled into bladder. As a source of ultraviolet light (UV) D--Light Storz xenon lamp (405 nm) was used. In 16 pts single PDD was performed in 38 double and in 97 PDD was performed three or more times (3 to 7). All pts were observed 12 months before and after PDD. In 102 from 151 pts in white light cystoscopy (WL) no changes in the bladder mucosa were found. In both kind of light (WL and UV) changes were not observed in 44 pts (29.1%). In 58 pts in WL no changes were observed but in UV places of neoplastic changes in the mucosa were found. In these cases the presence of superficial bladder tumours was confirmed. These changes were pathologically confirmed only in UV in 26 pts. Ta (G1 and G2) changes were confirmed in 52 pts (21 in WL and additionally 31 in UV). T1 changes were confirmed in UV in 26 pts but in WL in 25 pts. In 3 pts T2 changes were confirmed in both UV and WL. Cystoscopy in ultraviolet light (photodynamic diagnosis--PDD) with 5 amino levulinic acid gives more precise diagnosis of superficial bladder tumours then the one in standard white light. Cystoscopy performed to diagnose bladder tumours and early recurrence and new focus of tumours recognition without PDD is less effective. Introduction of superficial bladder tumours photodynamic diagnosis into health care system reduced recurrences and improves results of treatment. PMID- 15884238 TI - [Psychically ill patients from the view point of students from high schools, specialized in nursing education]. AB - The hypothesis that students from medical high schools present an open position to psychical ill patients was verified. There was made investigations through 30 of students from first year of Care and Teaching Health Class from Medical University of Silesia in Katowice. Students declared help and big interest for such kind of patients. We might see a big tolerance and acceptance through the investigated students. Probably, this kind of attitude was affected by media (TV, radio, press). Our results showed the ideal picture of students from nursery high school. On the other hand we have to think about the real attitude to psychical ill patients. Our future investigations during the next year of education will verify our preliminary results. PMID- 15884239 TI - [Significance of the DNA-HPV research in cervical cancer prevention]. AB - The purpose of our research was to assess DNA-HPV frequention observation and evaluation of the diagnostic value mRNA E6 and E7 HPV16 and HPV18 profile concentration in prognostic risk of intraepithelial lesions and cervical cancer progression in women with cytological screening. Human papilloma virus (HPV) infection were detected in 13.8% normal samples, presence HPV16 and 18 in 7.5% samples were detected. HPV 16, 18 or HPV16 and 18 infection were detected in 85% HSIL (high-grade squamous intraepithelial lesion) samples. HPV16 or HPV18 infection in 100% cancer samples were detected. In samples from control group expression of E6 and E7 genes were not detected. In LSIL (low-grade squamous intraepithelial lesion) group HPV16 E7 gene in 2.6% samples, in HSIL group E7 gene in 9.5% samples were detected. In all cancer samples E7 or E6 HPV16 and/or HPV18 were detected. PMID- 15884240 TI - [Prophylactic medical investigations for breast cancer and uterine cervix cancer in Regional Oncology Centre of Bydgoszcz in 2003]. AB - Kujawsko-pomorskie province is a region with high morbidity and death rate of breast cancer and uterine cervix cancer. Prophylactic medical investigations are methods to find less advanced forms as possible. The aim of the study was to estimate the results of prophylactic medical investigations of Regional Oncology Centre in Bydgoszcz in 2003. Mammography investigations were undertaken in 3 forms. The first one--so called "mobile"--was localized in chosen health service centres, the second one in "mammo-bus" reaching distant communes and the third one local outpatient clinics. Cytological investigations were rated by Bethesda and Papanicolau system. 23 923 people were investigated for breast cancer (age 35 79) and 7946 ones for uterine cervix cancer (age 30-74). 9183 medical investigations were taken by "mobile" device, 4592 in "mammo-bus" and 10 148 locally. 11 986 people had breast examinations, 6118 had gynaecologic examinations. There were cytological investigations results: I group--3902, II group--4019, III group--21, IV group--3, V group--1. Ten uterine cervix cancers were diagnosed (0.13% of investigated population). Five of them were histologically recognised as: carcinoma planoepitheliale praeinvasivum. 91 cases of breast cancers were diagnosed (0.38% of investigated population). Five of them were histologically recognised as: carcinoma intraductale typus comedo (there were: 3 simplex mastectomy, 2 tumorectomy with radiotherapy). The treatment of invasive cancers included 9 breast-conserving treatments. CONCLUSIONS: Prophylactic medical investigations let us find the cancer in early development stage and a number of cancer detections is a justification of this project continuation. The possibility to examine women from villages should have the influence on the detection of early breast and uterine cervix cancer. PMID- 15884241 TI - [Skin cancer screening program in the population of Bydgoszcz]. AB - The results of prophylactic medical skin examinations in inhabitants of Bydgoszcz were estimated. A prophylactic skin examination in asymptomatic patients was performed, and the most suspicion lesions were selected to excision. 750 persons were examined (age 15-93 years). 133 persons were operated. 173 skin lesions were removed. 32 skin cancers (18.53%), 3 melanomas (1.73%) and 34 skin lesions (which are base for melanoma) were confirmed. Detected carcinomas and melanomas were in early state of development. Prophylactic medical skin examination enable the detection of skin neoplasms in early state of development. Percent of detected carcinomas and melanomas of skin justify continuation this kinds of screening. PMID- 15884242 TI - [Physical breast examination]. AB - The aim of the project was the comparison of the effectiveness of physical breast examination in relation to mammography on the basis of research analysis conducted within the program "The Early Diagnosis of Breast Cancer". The program was conducted at J. Sniadecki Hospital in Nowy Sacz in the year 2002. The physical examination was carried out by professional midwives. The documentation of 400 women who participated in the program was analyzed in order to gain the necessary information. Half of the participants have identified anomalies in breasts (202; 50.5%). There was no difference in the number of the mentioned anomalies between the group of the younger (55; 50.0%) and older women (147; 50.7%). One of the three anomalies diagnosed in mammography was earlier diagnosed in the physical examination. It is important to indicate that it seems more difficult to detect the anomalies in the group of younger women (13; 23.6%) than in the group of the older women (47; 32.0%). Among all anomalies diagnosed in mammography more than half (98; 53.3%) are mastopathies. Only one of the five mastopathies (19; 19.4%) was detected in physical examination. The majority of women who participated in the prevention program live in the city (244; 61.0%). Women from cities had more breast anomalies (53.3%) than women from the country (46.1%). PMID- 15884243 TI - [Immunologic activity in coronary heart disease and atherosclerosis of the lower extremities]. AB - Cytokine activation may be connected to increase of clinical symptoms both of coronary heart disease and lower limbs atherosclerosis. Our aim is to determine the influence of the atherosclerosis generalization upon immune activation in coronary heart disease, with regard to ECG stress test. 127 patients have been included in the study: 21 with stable angina and peripheral artery disease (PAD)- group A, and 106 with stable angina--group B. 20 healthy persons comprised the control group (group K). The serum concentration of TNF alpha, sTNFR 1, E selectin and sVCAM-1 has been measured before and after the ECG stress test, using the ELISA method. Serum concentration levels of TNF alpha (A: 17.8 +/- 6.2 pg/ml, B: 17.4 +/- 3.8 pg/ml) and sTNFR 1 (A: 1678.5 +/- 600 pg/ml, B: 1376.4 +/- 558 pg/ml) have been significantly higher in both research groups than in the control group K (8.3 +/- 1.4 pg/ml, p < 0.001; 1093.9 +/- 457 pg/ml, p < 0.01). The sTNFR 1 concentration has been significantly higher in group A than in group B (p < 0.05). A significant post-exercise increase in E-selectin serum concentration has been observed, regardless to coincidence of lower limbs atherosclerosis. The atherosclerosis generalization level, e.g. the prevalence of PAD, in patients with coronary heart disease has an influence on immune activation--patients with lower limbs atherosclerosis are characterized by higher sTNFR 1 serum level. The ECG stress test induces the increase of E-selectin serum concentration in coronary patients, regardless of PAD. PMID- 15884244 TI - [Preparatory training of primary health care nurses for handling health education regarding prophylaxis of circulatory disorders]. AB - Circulatory disorders (CD) exhibit high incidence rate. At the same time the increasing number of data indicate that most of these disorders can be prevented. Prophylaxis is one of the professional functions of nurses. The preventive tasks included in this function are of special importance in achieving the goals of the National Health Programme and international CINDI WHO Programme--a model programme for the prophylaxis of chronic diseases and health promotion. The necessity exists to enhance the preventive actions, of which health education is an integral part. Therefore, it seems essential to prepare the nurses to conduct health education in the field of circulatory disorders. The aim of the study was to answer the following questions: 1. Do the primary health care nurses think that they are sufficiently prepared to conduct health education among their patients on the prophylaxis of circulatory disorders? 2. What sources do they use to obtain the knowledge about the prophylaxis of circulatory disorders? 3. Do they understand the need of getting and additional training in the prophylaxis of circulatory disorders and to what extent? The research problem was to determine whether there is a difference in responses between two groups: 1. nurses with secondary education and specialization and 2. nurses without specialization. The study involved all the nurses employed in the primary health care centres in Bialystok. Questionnaire was the research tool. 172 questionnaire forms were distributed, of which 146 were properly filled in. The study shows that the nurses with secondary education and specialization declared preparation to conduct education concerning the prophylaxis of circulatory disorders significantly more often than the nurses with secondary education without specialization. Worth noting is the fact, that the nurses with secondary education without specialization uttered a smaller need of additional training than those with secondary education and specialization. Nurses should be made aware of the necessity to enrich the knowledge in the field of methods, didactic materials and interpersonal communication in order to make health education more effective. PMID- 15884245 TI - [Attitude toward sexual life of young adult medical and nursing students]. AB - Nowadays can observe not only changes of views on human sexuality but also of its nomenclature. These changes also apply to the issues related to human sexuality which plays an role in psychological purposes. The article discusses psychological and pedagogical aspects of human sexuality on the threshold of adult life. 83 students of post-GCSE faculties of the medical school were interviewed. The aim of the study was to determine the attitude towards sexual life expressed by young people entering their adult life. The study tool was a questionnaire technique. Results were statistically analyzed. The results showed that: the level of knowledge of sexual life is low among adults: lack of correlation between emotional and cognitive elements of attitudes towards human sexuality; sexual behaviours depend on human sex. PMID- 15884246 TI - [Does coffee drinking influence serum uric acid concentration?]. AB - The drinking of coffee, a commonly used beverage, was a subject of many studies, mainly regarded to coffee influence on cardiovascular system. However, only one study indicates that coffee drinking in male adults may lead to decrease in serum uric acid level. Hyperuricaemia is a risk factor of many diseases. The aim of this study was to examine the influence of coffee drinking on serum uric acid concentration. 1955 working persons aged from 18 to 65 years were included into research. There were 571 women among them. We determined energy expenditure during professional work, blood pressure, body mass index, and measured serum levels of uric acid, glucose and creatinine. The amount of coffee and ethanol consumption was evaluated on the ground of an interview. It was showed that persons drinking coffee have lower serum uric acid concentration than non drinkers, especially among women, who drank more coffee then men. Uricaemia was correlated negatively with number of cups of coffee consumed and positively with body mass index, ethanol consumption and diastolic blood pressure. The author conclude that: 1) among women drinking on an average 10 cups of coffee per week appeared a decrease in serum uric acid concentration and a lower risk of development of hyperuricaemia, 2) elevated serum uric acid concentration is accompanied by elevated blood pressure and increased body mass index. PMID- 15884247 TI - [Prevention of viral hepatitis B and C infections--jurisdiction and certification difficulties]. AB - The number of the cases for payment because of viral hepatitis B and C inflammation is significant. Because of the greater patients' notice of the disease and portal of infection, lawyer's opinion accessibility and poor economical situation of society the claims for damages seems to be more popular. The lawyers specialize in medical law. This is the reason to pay more attention to prevention of infections. Besides the hospital infections there is problem of viral hepatitis B and C infection as the occupational disease with all health and juridical consequences. These problems concern specially operative specialties therefore gynecology and obstetrics among the others. The aim of our study was to analyze jurisdiction and certification difficulties in context of occupational disease and cases for payment because of viral hepatitis B and C infections. We also try to answer the question what are possibilities to prevent infections and to defence in law suits. PMID- 15884248 TI - [Basic methods in the prevention of breast cancer--diagnostic radiography]. AB - Breast cancer is one of the most common neoplasms of women in Poland. The study presents the diagnostic possibilities for early detection of neoplastic lesions and their differentiation from the radiologist point of view. Advantages and restrictions of individual methods and current views on prophylactic and diagnostic algorithms have been considered. The study has been based on long-term observation of patients coming to Breast Diseases Outpatient Clinic as well as on scientific reports. The importance of cooperation between radiologists and physicians with other specializations who deal with breast diseases has been stressed. PMID- 15884249 TI - [Selected conditions influencing the frequency of prophylactic gynecologic visits]. AB - The aim of the study was to estimate the frequency of prophylactic visits at gynaecologist among women and its correlation with: age, place of living, material status or kind of gynaecological illness. The study was carried out among 200 patients with different gynecological illnesses operated on in Department of Reproduction and Andrology University School of Medicine in Lublin. The study was carried out on the basis of author's questionnaire interview and analyses of medical documents. Participation in this study was voluntary and anonymous. The result showed that, 52% of women declared their regular (every half year and every year) visits at gynaecologist. Regularity of prophylactic visits depended on age, in favour of women under 35 year. The frequency of control visits was significantly higher among women living in the city. The majority (88.2%) of patients in the group of continuous medical therapy was the ones suffering from infertility. The education and material status did not influence the regularity of prophylactic visits at gynaecologist. PMID- 15884250 TI - [Cervical pregnancy--conservative treatment; case report]. AB - Cervical pregnancy is a rare form of ectopic pregnancy. This disorder appears with frequency 1 to 10 cases of ectopic pregnancy. For the sake of high risk of unexpected massive haemorrhage from cervical vessels, the cervical pregnancy is a life threatening state. Total hysterectomy was the only one method applied in this case recently. Since this procedure led to infertility, many clinical centers try to apply conservative treatment in nowadays. Haemostasis after evacuation of fetus from the cervix and early diagnosis of cervical pregnancy are the key to success of conservative treatment. In our study we present case of 24 years old woman with diagnosed early cervical pregnancy and subjected to conservative treatment. PMID- 15884251 TI - [The effect of transdermal hormone replacement therapy on vasomotor symptoms in perimenopausal women]. AB - About 30% of female population in Poland is already in the perimenopausal age. The North American Menopausal Society has defined the menopause as the permanent cessation of menstruation resulting from the loss of ovarian follicular activity. Perimenopausal period is often connected with vasomotor symptoms in women. In recent years, there has been a growing recognition among clinicians of the importance of learning more about how patients cope with the symptoms of their condition. The aim of the study was to evaluate the effect of hormone replacement therapy on vasomotor symptoms in perimenopausal women with the use of Kupperman Index. In a six-month (6 treatment periods, 28 days each) randomized study, 75 healthy postmenopausal women were assigned to receive 50 micrograms/day of continuous transdermal estradiol with sequential transdermal norethisterone acetate (NETA) in daily doses of 170 micrograms in a single transdermal patch. The average number of vasomotor symptoms per day measured with Kupperman Menopausal Index decreased from prestudy by over 90%. The substantial improvement in vasomotor symptoms assessed across time was apparent by week 12 of the treatment. PMID- 15884252 TI - [Hormone replacement therapy--oral and transdermal administration]. AB - Nowadays almost one third of women's life follows the cessation of menstruation. Therefore medical investigations are concerned with the improvement of women's quality of life in this period. Hormone Replacement Therapy is one of the possibilities of restoration women's premenopausal hormonal status. In the paper advantages and disadvantages of oral and transdermal administration of Hormone Replacement Therapy are presented. PMID- 15884253 TI - [Late maternity in researchers' and medical staff's opinions]. AB - During the last decade in Koszalin hospital 223 women over 35 years old gave birth to their first child. Those numbers made up 1.2% of all births in this time. Medical documents were analyzed to compile a detailed list carrying main problems concerning of this group of women. Older primiparas had a lot of problems with conception, carrying pregnancy to term, pathology of pregnancy and labor. These women cooperated with midwives and doctors very well, and that fact was strongly emphasized by the medical staff. Women over 35 years old declared stronger need of support, aid and knowledge than other groups of patients. PMID- 15884254 TI - [Evaluation of medical instructions designed by hospitals for expectant mothers]. AB - The proper prenatal care decreases the risk of morbidities and complications during the pregnancy. The high level of the gestational knowledge supplied to woman and her family, permits to recognize the signs of threats and immediately go to a specialist and get proper help. A women consciousness of the proper gestation hygiene simplifies to avoid the risk of fetal development disorders. Because of this, the medical personnel is obligated to promote good behaviors adequate for that period. The pregnancy is a condition in which the salubrious promotion is most effective because at this time women and their families are disposed to change the wrong health behavior. The object of the study was to assess the health education process among pregnant women. One hundred women after the delivery was examined by means of self-created questionnaire. None of the required issues was properly discussed with the pregnant women. PMID- 15884255 TI - [Analysis of women nutritional status during pregnancy--a survey]. AB - The proper diet is one of the most important factor during pregnancy. The general knowledge about proper nourishment during pregnancy allows the women to avoid quantitative and qualitative nourishment mistakes. Because of this--the salubrious education in this aspect is very important. The aim of the study is to analyze the proper nourishment during pregnancy particularly in professionally active women and those who don't work during pregnancy. PMID- 15884256 TI - Endometrial ablation--long-term complications. AB - Endometrial ablation is the alternative of hysterectomy performed to treat dysfunctional uterine bleeding in premenopausal women. Although early complications are significantly documented, little is known about their late complications. The aim of the study was to evaluate long-term complications of endometrial ablation. We audited two hundred and ten patients (mean age of 43) to assess the effectiveness of alternative of hysterectomy (endometrial ablation) in long term. This article presents four patients who presented with acute abdominal pain a few years after successful endometrial balloon ablation. PMID- 15884257 TI - [Husband's presence at childbirth in light of obstetric psychoprophylaxis]. AB - Contemporary obstetric psychoprophylaxis gives prospective parents wide opportunities to prepare to the pregnancy period and delivery. It is educationally-minded and points the importance to modify the life style, introduces exercises accompanied by the relative during the pregnancy and delivery. The survey portrays husband's--child father's role in obstetric psychoprophylaxis. The importance to continue the psychoprophylaxis in the delivery room was spotted in the survey, too. The continuation might be reached by close relative's presence. PMID- 15884258 TI - [Sexual behaviors of pregnant women]. AB - Pregnancy and childbirth are both the part of woman's sexual life. Changes occur in every trimester of pregnancy have significant influence on the sexual behaviours. The purpose of our monograph was the analysis of behaviours of pregnant women. We comprised intensity and effectiveness of the sexual relations in each trimester of pregnancy. We analysed life circumstances affects on the sexual behaviour (own bedroom) and sexual life in every trimester of the pregnancy in compliance with: libido, prevalence of orgasm, sensitivity and erotic imaginations. The inquiry concerned the sexual positions preferred by pregnant women too. We paid respect on the subjective attractiveness evaluation in pregnancy and on fear of sex in pregnancy. We found some differences in every trimester of pregnancy refer to libido and orgasms frequency with particular reduction of these phenomenon's in third trimester. The increase of both: sexual imaginations and fear for sex was found. The fear for sex in pregnancy mainly concerns the appearance of bleeding or miscarriage. PMID- 15884259 TI - ["Heart attack in humans"--the health belief model in comparison with the health reality of patients with myocardial infarction]. AB - Myocardial infarction (MI) is the direct cause of 40% of all deaths, independently of the high standard of medical treatment. The main aim of the study was to define the relationship between the declarative health model (declarations) and the real health activity of the sufferers (realization). The research was performed on 107 patients after acute MI, hospitalized in the Cardiology Department Medical University of Silesia. They were questioned in direct interviews. In addition, the modified form of Shalit's circle was applied. The results indicated the coexistence of two health models of patients. In declarations, there is a model of person with interior attribution of MI (90.65%) that has a feeling of ability to influence his/her health (e.g. through the lifestyle: 63.55%; an individual's character: 41.12%). In realization, there is a model of sick patient with exterior attribution of heart attack (e.g. political and economic crisis, innate predisposition). A similar tendency in the perception of health factors and planning of changes after MI was observed. For instance, 50.47% patients talked about "the increase caution" or "slowing down" as a new planned form of activity. Moreover, of 92% patients who have the social support only 33.64% consider that as an important factor for health. The results suggest a divergence between declarations and real health activity. The launched health model of beliefs seems to be too costly and not attractive enough. Taking into consideration the complete risk factors in MI and making the proposed health belief system more accessible seem to be essential for creating an adequate prevention program. PMID- 15884260 TI - [Family planning--the role of general practitioner in abortion prophylaxis]. AB - According to World Health Organization, abortion is defined as an induced termination of pregnancy by use of medications or surgical interventions after implantation of the embryo and before the fetus is able to survive outside the maternal organism (before 22nd week of pregnancy). More than 75 millions of women experience unwanted pregnancy every year. Contraception for that group was either unavailable or the information about contraceptives use possibility was not efficient. Lack of conversation about family planning with the partner, rapes and inefficiency of contraceptives (8 to 30 millions women a year) might be other reasons. More than two-third such pregnancies are terminated by abortions. The number of women in reproductive age (15-44) is 1.38 million. Most of them are sexually active but not willing to have progeny. The decision of abortion is taken every year by 35 per 100 females (26 millions of legal abortions each year, 20 millions of illegal). By 1986, 36 countries introduced liberal abortion law that gives permission for abortion only for social, medical and personal reasons. The main law regulating the permission of abortion in Poland is a resolution of Family planning, embryo protection and conditions for conducting pregnancy termination from 1993, modified in 1997. In 1999, 151 abortion procedures were performed and that number is decreasing gradually. The development of so called "abortion basement", where unsafe abortion is usually performed, is a consequence of restrictive policy about the abortion law. In the last few years the holistic and individual approach to the patient has started to play an important role. Unfortunately, in the case of sexual education and knowledge of conscious family planning, medical services, and medical doctors especially, play only the minimal role. It seems to be essential in gynecological and general medicine practice to give information about different methods of family planning and protection against sexual transmitted diseases as well as to control patient's health when using contraceptives. PMID- 15884261 TI - [Osteoporosis--pathogenesis and prophylaxis]. AB - Osteoporosis is defined as a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue with a subsequent increase in bone fragility and susceptibility to fracture. In this paper some facts concerned with pathogenesis and prophylaxis of osteoporosis are presented. PMID- 15884262 TI - [Influence of GnRH analogue on the intensification of endometriosis symptoms and infertility treatment]. AB - OBJECTIVES AND STUDY DESIGN: The aim of this study was to evaluate the influence of the treatment with oral doses of 50 mg GnRH analogue on the intensification of endometriosis symptoms and infertility amongst women with evident symptoms of endometriosis in comparison with placebo group. A group of 34 women at the age from 18 to 45 were introduced into the study. The inclusion criteria for investigated population contained: endometriosis symptoms, endometriosis diagnosed by laparoscopy, its surgical or pharmacological treatment, negative pregnancy test score and regular menses. The patients were divided into 2 groups: investigated group and control group. Women completed "Diary" every day in which they estimated main endometriosis symptoms: dysmenorrhea, dyspareunia, pelvic pain and vaginal bleeding (according to Pain Grading Scale). The intensification of dysmenorrhea and vaginal bleeding was reduced in the investigated group in comparison with control one. These differences were statistically significant. Although the extremity of dyspareunia was decreased in two groups, this correlation was statistically significant only in the investigated group. Pelvic pain evaluation showed that its level was lower in the investigated group (p > 0.05). It was found that 11 women (investigated group) and 5 women (control group) got pregnant after the period of 12-week treatment. CONCLUSIONS: GnRH analogues have an efficient influence on the reduction of endometriosis symptoms. GnRH analogues could be used in the management of infertility. PMID- 15884263 TI - [Analysis of obstetrical and gynaecological history of women with urinary stress incontinence]. AB - Urinary leakage is a hygienic and social problem, that concerns women both in reproductive and perimenopausal age. It leads to a life deterioration, depression, and social isolation. Urinary stress incontinence (USI) is one of the most common-type of urine leakage. Characteristic symptom of this disorder there is involuntary leakage of urine during increased intra-abdominal pressure (cough, laugh, sneezing or physical work). There are many various predictive factors of urinary stress incontinence in a large number of publications. One of them, the most important, seems to be a gravidity and labor. 40-82% pregnant women complain of urinary leakage. The main aim of investigation was to determine obstetrical and gynaecological predictive factors of urinary stress incontinence. 53 women with diagnosed USI and qualified to TVT procedure have to fill in questionnaire with row questions about their obstetrical and gynaecological history. Our investigation revealed that significant influence on appearance of USI exert number of spontaneous labors, birth weight >3500 g, lack of substitute hormonal therapy during menopause and hard physical work in the past. PMID- 15884264 TI - [Quality of life of women with urinary incontinence]. AB - The aim of the study was to estimate a quality of life of women treated for incontinence of urine. The study included 119 women hospitalized in Clinic of Gynecology of Medical University of Lublin for urine incontinence from January 2001 till September 2002. A questionnaire based on King's Health questionnaire was used as a tool. Data obtained from questionnaire prove a strong influence of symptoms of incontinence of urine (p < 0.05) not only on an estimation of health state, but first of all on daily duties and professional work of examined women. Symptoms of incontinence limit an interpersonal communication, participation in active rest, family life and sexual activity. About 2/3 (28.57%) of examined women estimate their psychic state as very bad, (31.93%) as bad and (32.77%) as a little poor and only few women (6.73%) state that problems with incontinence of urine have small influence on their emotional state. CONCLUSION: The appearance of urine incontinence symptoms have an influence on quality of life of women suffering from this disease. PMID- 15884265 TI - [Urinary incontinence in pregnancy]. AB - The aim of the study was evaluation of the frequency of urinary incontinence occurrence in pregnancy and the identification of the systemic and obstetric causes. The study was carried out on the turn of January to February among 146 women in childbirth staying at maternity wards of the Clinical Hospital and District Hospital in Lublin. It was stated, that incontinence problems in pregnancy were reported by 79.5% of the surveyed women. The symptoms of urinary incontinence in pregnancy were reported by 37.7% of women, and 38.2% of women declared to be unable to stop the stream of urine. The first symptoms of urinary stress incontinence occurred in 3.6% women before first pregnancy, in 12.7% women they occurred in the former pregnancy, and in 5.5% after previous delivery. In women with symptoms of urinary stress incontinence the BMI index was higher than in the rest of the studied group. This study revealed the correlation p < 0.05 between the occurrence of urine incontinence the profession (weight lifting). PMID- 15884266 TI - [Venosus thromboembolism in pregnancy--case report]. AB - Venosus thromboembolic disease is an important problem in pregnant women. The overall risk of venosus thromboembolism in pregnancy is 6-times greater then in non pregnant women. In the article we present a case of a pregnant woman with venosus thromboembolism. The analysis of the pregnancy and mode of delivery, development of disease, diagnostic and therapeutic method. PMID- 15884267 TI - [Parents' and caregivers' theoretical and practical knowledge of first aid in case of accidents and minor injuries in children]. AB - Accidents and injuries cause most serious health problems in pediatric group of patients in Poland. We asked a group of 93 parents and tutors (11 men and 82 women) to complete a questionnaire containing 20 questions. It referred to their knowledge on first aid treatment. We have analyzed the answers using statistical methods and couched our conclusions. 1. The results of the enquiry show a great extent of self-satisfaction in the questioned group of parents. Most of them (64.5%) think they know first aid rules, although only 35 people (37.6%) were ever trained in this field. Parents declare they known, how to deal with such conditions as bums, slight contusions and injuries, fractures, torsions, choking and high temperature. 2. Most parents (71%) have already treated minor injuries and dealt with minor accidents which took place while they took care of a child. Most common health problems were: high temperature--23.9%, slight contusions- 22.4%, minor injuries--16.1%, hemorrhage--8.8%, scalding--8.3%, choking--7.3%. Accidents occur mainly during playground activities or cycling (63%) and at home (17%). 3. When an accident involving children occurs, 29.2% of parents call for medical help, 63.8% of them do it only if very serious injuries occur and 9% always try to treat a child themselves. In our enquiry most parents knew the names of popular analgesic and antipyretic drugs. They also knew how to deal with a slight burn or scald. 4. In questioned adults' opinion health care professionals do not give enough first aid information to the society. In our examined group, 58.1% of parents try to read and learn themselves. This shows the need for training people who take care of children in first aid treatment. As the best places for such training enquired parents pointed schools, kindergartens, pediatric and obstetric clinics. PMID- 15884268 TI - [Some aspects of the influence of television on children in early adolescence]. AB - Television is treated not only as a carrier of information, but first of all as a source of entertainment. From the other sites, it also carries many threats to young spectator. It is known already today, that many remittances are full of aggression. In what way does this special group of audience uses the TV remittances? This problem is analyzed in presented study. Diagnosis of problem was conducted among children attending to secondary schools. An investigative tool was a questionnaire of inquiry which included following questions: quantity of time spent in front of television set, influence of television on functioning of a child, rules of using the television at home, what kinds of televisions' offers children use, influence of television on social functioning of child (contacts with peers, with family) and influence of television on intellectual functioning of a child. PMID- 15884269 TI - [Rehabilitation of patients after lower limb amputation as a basic element of adaptation to normal life]. AB - Amputations of bottom limbs are serious problem. Loss of leg causes always heavy psychical injury, it makes life more difficult, as well as moving and self service. The change of appearance and shape of body demands adaptations of patient and his neighbourhood. Every amputation is not only heavy physical injury but also violent and long-lasting psycho-emotional and social stress. Most of patients are afraid of unknown, so it's necessary to decrease their fear, they expect our help in solving their social and domestic problems. The aim of investigations is to show the process of adapting to normal life of people after amputation of bottom limb on example of their rehabilitation. The following investigative methods were used: steered observation, interview, Polish version of questionnaire RNL (reintegration to normal living) in small modification and inquiry. Investigations were conducted in two hospital wards and the rehabilitation centre. The group consisted of 82 patients--59 men (72% of group) and 23 women (28%). The numerous group (43%) were people between 50-69 years old. Only a few patients after amputation go to special out-patients' department. The main reason of it is the lack of rehabilitation centre in the neighbourhood. 1/5 of the group regularly uses rehabilitation at expert, however most patients over 70s do not use any rehabilitation. Frequent form of activity at studied people is morning exercises and easy exercises of stump. Lack of physical activity is typical for 1/3 of group. Physically active are only 12 patients (15%). Near half of the group uses artificial limbs, but 1/5 of group does not want to have it. Most of them is over 70 years old. Results of investigation show the need of opening the larger quantity of rehabilitation centres to make the rehabilitation more common and accessible. Essential meaning has also bigger motivation of patients to physical activity as well as using artificial limbs, which do facilitate functioning in everyday life and improve the mood of patients after amputation. PMID- 15884270 TI - [Value of selected biophysical surveillance methods in prediction and prophylaxis of complications associated with adverse pregnancy outcome in intrauterine growth restriction]. AB - Intrauterine growth restriction is associated with higher rate of complications occurring during pregnancy and in labour, increased morbidity and mortality rate in neonatal period, impaired psychosomatic development in childhood and increased susceptibility for particular diseases in adulthood. For improvement of perinatal outcome it is essential to establish objective criteria based on biophysical surveillance methods for prediction and prophylaxis of adverse outcome in IUGR pregnancies. The aim of the study was to compare selected biophysical surveillance methods in prediction and prophylaxis of complications associated with adverse pregnancy outcome in intrauterine growth restriction. There were statistically significant differences in CTG, BPS, AFI, and pulsatile index values in umbilical artery and MCA between IUGR and normal fetuses. The best predictor of adverse pregnancy outcome in IUGR fetuses was PI value in umbilical artery. PMID- 15884271 TI - Rheumatic heart disease in Ethiopia: could it be more malignant? AB - The objectives of the study were to evaluate severity of rhenumatic heart disease (RHD) using age at time of death as its indicator and to look for factors associated with the severity. Retrospective study of consecutive cardiovascular deaths in the medical wards of Tikur Anbassa Teaching Hospital (TAH) from January 1995 to December 2001 was carried out. Information on age at the time of death, sex, type of valve lesions, status of secondary prophylaxis, presence or absence of arrhythmia, infective endocarditis, embolic phenomena, co-morbid illness, immediate cause(s) of death and duration of follow up at health institutions were obtained There were a total of 457 cardiovascular deaths including cerebrovascular accidents (CVA). 121(26.5%) were due to RHD. Charts of 115 RHD patients were available for detailed analysis. The overall mean age at the time of death was 25.89+/-11.05 years. The median age for males was lower than that of females (19 years, I.Q. 16-32 versus 25 years, IQ 15 - 30 years), but not statistically significantly different (p = 0. 10). There were more female deaths accounting for 57.4 %. About 70% of RHD patients died from congestive heart failure. Eleven per cent (13 patients) each died from systemic embolism and co morbid conditions. Combined mitral and aortic valve disease within a patient accounted for majority (42.6%) of the lesions followed by combined mitral regurgitation and stenosis (24.4%). Isolated mitral stenosis, or regurgitation was a relatively less common cause of death. Aortic valve lesion without mitral valve involvement was a rare cause (3.5%) of death. In conclusion RHD in our patients ran more aggressive course as indicated by death occurring at much younger age than in even the preprophylaxis era in western world. The more frequent combined valve lesions and infrequent secondary prophylaxis may significantly contribute to the rapid course, but cannot fully explain the very low mean age. Factors responsible for rapid course in an Ethiopian need to be addressed urgently. PMID- 15884272 TI - Socio-demographic profile and obstetric experience of fistula patients managed at the Addis Ababa Fistula Hospital. AB - Obstetric experience, clinical and socio-economic characteristics, and reasons for preference of place of delivery of 639 fistula patients admitted to the Addis Ababa Fistula Hospital between May 1999 and February 2000 are described About 94% of fistula patients were married and 83.6% had been through with the delivery caused the fistula before the age of 20. The mean ages at the first marriage and at the causative delivery were 14.7 (sd=2.6) and 17.8 (sd=3.2) years respectively. Mean height of fistula patients studied was 149 cms (sd=8). About 64% were primiparous, 44% (279) delivered at home, and labor lasted for 3.8 days on average. Majority of fistula patients mentioned distance as a main problem for the delays. Lack of money, poor knowledge and delay in referral were the other frequently mentioned reasons for the delays. About 62%(399) of fistula patients owned nothing valuable. Of 279 patients delivered at home 186 were those who owned nothing. Out of 180-fistula patients mentioned distance as the major problem, 103 (57%) stayed at home for delivery. About 54% of fistula patients were already divorced on arrival to the hospital and this figure varied among women with different parity and among those owning different property. Teenage and short stature are observed features of fistula patients. The study also showed the huge problems faced by fistula patients in accessing emergency obstetric care services. Analytic study is recommended to compare the magnitude of these problems among other mothers with different obstetric outcome. Meanwhile, community education about problems following teenage pregnancy, sign and symptoms of obstructed labor and the advantage of institutional delivery might reduce the occurrence rate of obstetric fistula. Community organized fund and maternity waiting areas for young and short expecting mothers are among other recommendations to be considered. PMID- 15884273 TI - Clinical profile of acute renal failure in children admitted to the department of pediatrics, Tikur Anbessa Hospital. AB - Case records of 30 pediatric patients with the diagnosis of acute renal failure (ARF) admitted to Tikur Anbessa Hospital in Addis Ababa between October 1997 and October 2001 were analyzed There were 15 females and 15 males. Three patients had glomerulonephritis, in two patients the cause of acute renal failure was not known, one child had obstructive uropathy. Twenty-three patients had post diarrheal hemolytic uremic syndrome. The age ranges of post-diarrheal hemolytic uremic syndrome cases were between 0.6 years and 7 years with a median age of 2.2 years. Fourteen patients died of acute renal failure among this hemolytic urmic syndrome contributed to the death of 9 patients. The commonest clinical presentation was severe oligo-anuria in (25 patients), edema in (22 patients), and bloody diarrhea in (21 patients). From stool cultures of 16 patients with hemolytic uremic syndrome, there were five isolates of Shigella species, two isolates of E. coli, and two isolates of Salmonella species. Five patients had non-oliguric acute renal failure. Hemolytic uremic syndrome is the leading cause of acute renal failure in infants and young children in our series. Vigorous resuscitation and early dialysis will reduce mortality rate. PMID- 15884274 TI - Retrospective analysis of 10 year medical board proceedings at Amanuel Hospital Addis Ababa, Ethiopia, 2001. AB - The Medical board at Amanuel Psychiatric Hospital assesses mental disability. Analysis of the board's proceedings from 1985--1994 is discussed in this paper. The proceeding record showed that the board has assessed 1963 during the ten-year period, but analysis was done only on 1854 cases because of incomplete information. Over 80 % of the cases assessed by the board were males and 96% were below the age of 50 years. Teachers were presented to the board more often than other occupational groups. Over 50% of the cases presented to the board had psychotic illness and 12% had no mental illness. There was a decline in the number of cases presented to the board in the later part of the ten year period (Chi-square for trend=206.36, P<001). Request for assessment of fitness accounted for 79% of all requests presented to the board. Positive recommendation to the request was given to 66% of the cases and transfer from one working place to another was the most frequently given recommendation. It appears that assessment of disability used to be one of the major activities of the psychiatrists at Amanuel Hospital. Creating conducive working environment to the employees, putting some effort to change the attitude of employers toward the mentally disturbed worker and training health workers working in the clinics of enterprises/organizations on basic counseling principles are recommmended to minimize the flow of cases to Amanuel Hospital for this purpose. PMID- 15884275 TI - In vivo sulphadoxine-pyrimethamine sentitivity study Tigray Region, Southern Zone, Alamata Town, September--November 2001. AB - The in vivo therapeutic efficacy of sulfadoxine- pyrimethmanine (SP) was assessed in a clinical setting involving 77 falciparum malaria patients diagnosed at Alamata Sector Laboratory, Southern Tigray in Sept--Nov. 2001. The objective of the study was to ascertain the continued usefulness of SP in the routine treatment of uncomplicated falciparum malaria. The patients were selected among 370 patients, aged 6 months and above, presenting for evaluation of febrile illnesses. The 1996 World Health Organization protocol for assessment of therapeutic efficacy of anti-malarial drugs was employed to select patients fulfilling enrollment criteria. Adequate Clinical Response (ACR) was detected in 75 patients (97.4%) whereas Late Treatment Failure (LTF) was ascribed to the remaining 2 patients ( 2.6% ) This data indicates that SP must have been effective as a first-line treatment for uncomplicated falciparum malaria in Alamata during the survey in 2001. Notwithstanding, the need for continuos surveillance on the therapeutic efficacy of SP as well as the in vitro resistance of P. falciparum is recommended. PMID- 15884276 TI - Nine-year trends in HIV-1 prevalence among visa applicants in Urban Ethiopia. AB - A retrospective study was conducted to examine trends in HIV-1 prevalence among visa applicants between the years 1993 to 2001 in Urban Ethiopia. A total of 63,869 visa applicants were screened during these nine years period. The majority of them (79.5%) were females. Their mean age was 31.6 and 25.7 years for males and females, respectively. HIV-1 prevalence ranged from 6.8% in 1993 to 10.4% in 1997 (test for trend: p<0.001), while it seems stabilized at around 11% after 1997. The overall period prevalence was 9.5%. The peak prevalence was documented in the age group 25-29 for females (12.1%) while it was in the age group 30-34 for males (11.4%). This study, therefore, confirms the severity of the HIV-1 epidemic in the country. Visa applicants can be used as a sentinel population for monitoring trends in HIV-1 prevalence in the country, although additional socio demographic information would be useful for better interpretation of such data. PMID- 15884277 TI - Primary extradural calvarial meningioma: case report. AB - Intracranial meningiomas usually develop intradurally with in the confines of the skull. Meningiomas originating in an extradural location are rare. A woman of 69 years had a hard mass located in the left parietal region. The neurological examination and laboratory data were with in normal limits. Plain skull X-rays revealed hyperostotic changes. The meningioma was removed in bloc. And histologically, it was found to be transitional type of menigioma. PMID- 15884278 TI - Torsion of a pelvic wandering spleen as a cause of acute abdomen in a woman: a case report and review of the literature. AB - Wandering spleen is a rare clinical condition in which the spleen is devoid of its normal peritoneal attachment associated with a long vascular pedicle. Women between the ages of 20-40 years are more likely affected. The common clinical presentation is abdominal mass with pain. For preoperative diagnosis a high index of suspicion is helpful. Ultrasonography and computerized tomography confirms the diagnosis. Treatment is surgical, splenopexy being the most appropriate. If ischemia is persistent after detorsion splenectomy can be done either laparoscopically or through laparotomy. A 25 year old female patient with torsion of an ectopic spleen presenting as a case of acute abdomen is presented. Review of the literature is done and presented PMID- 15884279 TI - Assessment methods in medical education. AB - Assessment is a component of any instructional system. In medical education several methods have been in use and newer techniques are emerging depending on specific curricula. In using any assessment method it is important to check for practicality, validity and reliability. The methods that are being used currently in Ethiopia have been in use in many countries and still remain as the predominant methods used in many parts of the world. The methods of assessment used in Ethiopia include long case, short case, viva voce and progressive assessment. Each of these methods have its own weaknesses and strengths. The long case has high face validity though there are many advantages related to patient selection and patient and examiner variation, resulting in low reliability. The specific advantage of short case is the diversity of patients observed by the candidate though subjectivity and inter rate variability remain major drawbacks. Progressive assessment is useful in that direct observation and more exposure are used to assess students. However, subjectivity remains a major weakness of this method. It is noted that viva voce is affected by subjectivity and personality of the candidate and appears to add little to the other methods of assessment. In general, the relationship between the various methods of assessment is not consistent. There is a need to find out more objective and effective methods of assessment or at least, improve on the existing ones. In the mean time, in using any of these assessment methods, one should look into the weaknesses and strengths of each method. PMID- 15884280 TI - Coverage of scientific research by the news media. PMID- 15884281 TI - Mycobacterial adenitis: role of Mycobacterium bovis, non-tuberculous mycobacteria, HIV infection, and risk factors in Arusha, Tanzania. AB - OBJECTIVE: To assess risk factors and mycobacterial agents in mycobacterial adenitis. DESIGN: Cross sectional involving comparison analysis of high-risk groups. SETTING: Seven hospitals in rural and semi-rural districts of Arusha. SUBJECTS: The study comprised of 457 patients of clinically diagnosed mycobacterial adenitis. INTERVENTIONS: Biopsy materials were cultured and identification of mycobacterial isolates, and HIV infection testing were performed using standard methods. A questionnaire was used to establish information for assessing risk factors. MAIN OUTCOME MEASURES: Proportions of mycobacterial isolates, risk factors and odds ratios. RESULTS: Of the 457 specimens, 65(14.2%) were culture positive. Isolates identified were M. bovis, 7(10.8%) M. tuberculosis, 27(41.5%) and non-tuberculous mycobacteria 31(47.7%). HIV infection and ingestion of raw milk were linked with increased risk of M. bovis infection by OR of 13.6 (95% CI, 1.7 - 109.9) and 15.28 (3.26 - 71.7), respectively. On multivariate analysis, an OR of 16.2 (1.3 - 201.3) for having M. bovis adenitis was linked to HIV infection, raw milk and houses with poor ventilation. An OR of 5.2 (1.2 - 20.6) for non-tuberculous mycobacterial adenitis was linked to history of TB in the family, HIV infection, raw milk, raw animal products and poor knowledge on transmission of tuberculosis. CONCLUSIONS: M. bovis caused one out of ten cases of culture positive mycobacterial adenitis. Non tuberculous mycobacteria were more common than M. tuberculosis (50% and 40% of the cases, respectively). HIV infection and raw animal products are among the risk factors identified for M. bovis and non-tuberculous mycobacterial adenitis. PMID- 15884282 TI - Comparison of vaginal and oral misoprostol, for the induction of labour in women with intra-uterine foetal death. AB - OBJECTIVE: To compare the efficacy of vaginal and oral misoprostol for the induction of labour in women with intra-uterine foetal death (IUFD). DESIGN: A prospective randomised clinical trial, comparing 200 microg oral and 200 microg vaginal misoprostol, six hourly for a maximum of four doses for the induction of labour in women with IUFD. SETTING: Ga-Rankuwa hospital (Department of Obstetrics and Gynaecology), Pretoria, South Africa. It is a tertiary institution serving predominantly black indigenous population. MAIN OUTCOME MEASURES: The primary outcome measure was the induction to delivery time, and secondary outcome measures were the number of patients requiring augmentation with oxytocin and all complications were noted. RESULTS: Twenty women were randomised to the vaginal route and 18 to the oral route. The induction to delivery time was shorter with vaginal misoprostol (13.5 +/- 8.3 hrs) compared to oral misoprostol (21.4 +/- 13.9 hrs; p < 0.05). There was no significant difference in the amount of misoprostol needed to achieve successful induction in the two groups. More women (10/18) who received oral misoprostol required oxytocin augmentation to complete the induction of labour compared with 4/20 women in the vaginal group (p < 0.05; Odds Ratio 2.8; 95% Cl 1.36 - 4.24). There were no cases of failed induction. The systemic side effects (shivering, diarrhoea, vomiting and pyrexia) were more common with oral misoprostol (44.5%) compared to vaginal misoprostol (20%). This difference gives an overall Odds Ratio of 2.2 at 95% Cl of 1.6-2.8(p < 0.05). CONCLUSION: Vaginal misoprostol achieved successful induction of labour in women with IUFD in a shorter time than oral misoprostol with significantly less side effects. PMID- 15884283 TI - Prevalence of hepatitis A, B, C and human immunodeficiency virus seropositivity among patients with acute icteric hepatitis at the Kenyatta National Hospital, Nairobi. AB - OBJECTIVE: To determine the prevalence of hepatitis A, B, C and HIV seropositivity among patients with acute icteric hepatitis. DESIGN: Cross sectional descriptive survey. SETTING: Kenyatta National Hospital, Nairobi. SUBJECTS: Eighty four patients aged above six months with a history of jaundice not exceeding six months were recruited. There were 47 males and 17 females with an age range of eight months to 67 years and a median age of 25 years. METHODS: History was obtained physical examination done and blood taken for determination of bilirubin, ALT, AST and ALP levels. Sera that had disproportionately greater transaminase than ALP elevation were assayed for IgM anti-HAV, IgM anti-HBc, HbsAg, anti-HCV and anti-HIV antibodies. RESULTS: Evidence of hepatitis A, B, and C was round in 41.7%, 26.2%, and 7.1% of the patients respectively, 13.1% of the patients were HBsAg carriers while 30.1% of all patients were HIV positive. Thirty two patients did not have evidence of hepatitis A, B, or C infection and this group was significantly associated with HIV infection (p = 0.003). CONCLUSION: Hepatitis A was the commonest overall type of acute icteric hepatitis seen at the KNH, and among patients aged 15 years and below. Hepatitis B was the leading identified cause of acute hepatitis among those aged over 15 years. Hepatitis C accounted for 7.1% of acute icteric hepatitis 30.1% of all patients and 50% of those admitted with acute hepatitis were also HIV positive. PMID- 15884284 TI - Anorexia nervosa in Kenya. AB - BACKGROUND: Anorexia nervosa is a rare disorder in Africans, inspite of posing a serious public health hazard in the West. Whereas it is possible that African psychiatrists lack the skills to diagnose the disorder, other possible explanations for its apparent rarity must be sought in view of emerging evidence, which suggests a real lack of occurrence. OBJECTIVES: To establish the knowledge of practicing Kenyan psychiatrists about the signs, symptoms and management of anorexia nervosa and to establish the number of cases they had seen during all their years of practice of psychiatry. DESIGN: Telephone survey and data collection: case-finding approach. SETTING: Kenya, June 2001 METHODS: Forty seven psychiatrists registered to practice in Kenya as of June 2001 qualified for inclusion. The respondents were interviewed on the same day, to reduce the likelihood of cross checking between colleagues. RK called each one, (all knew her) explained the nature of the survey and obtained verbal consent to administer the brief (10 minutes) questionnaire, developed by the authors for the purpose. RESULTS: Twenty seven of all those eligible were reached on the day. All but one gave consent for the interview (55% response rate). The 20 who were not reached were similar to the respondents with regard to experience in psychiatry, (mean duration 11.4 versus 10.9 yrs) but differed in their place of residence, the majority of those not reached resided out of Nairobi (60% versus 26%). In a cumulative total of 320 years of practice, they had seen 16 cases of which seven were of African origin. The rest were Caucasian or of Asian origin. The psychiatrists demonstrated adequate skill in recognising anorexia nervosa. CONCLUSION: Kenyan psychiatrists can recognise cases of anorexia nervosa. The condition is rare in Kenya. The reasons for this remain unclear and traditional explanations for its cause as due to pressure for thinness may not be adequate for the Kenyan case. PMID- 15884285 TI - Gastrointestinal injuries from blunt abdominal trauma in children. AB - OBJECTIVE: To determine the pattern, presentation and outcome of gastrointestinal injuries from blunt abdominal trauma in children. DESIGN: A retrospective study. SETTING: Ahmadu Bello University Teaching Hospital, Zaria, Nigeria. SUBJECTS: Twenty one children managed for gastrointestinal injuries from blunt trauma from 1984-2002. MAIN OUTCOME MEASURES: The pattern, presentation, management and outcome of gastrointestinal injuries from blunt trauma. RESULTS: In the 19 year period, 1984-2002, 92 children were treated for blunt abdominal trauma, 21(23%) of who had injuries to the gastrointestinal tract. Three presenting after 24 hours had evidence of peritonitis. In six children with isolated gastrointestinal tract (GIT) injury who presented within two hours, abdominal signs were vague at initial evaluation but became marked over a few hours at repeated examination. In eight with associated intraabdominal injuries, abdominal signs were marked at initial examination and five presented with shock. Free peritoneal air was present on plain abdominal and chest radiograph in three of ten patients, dilated bowel loops in six and fluid levels in one. Diagnostic peritoneal lavage or paracentesis was positive in four patients with isolated GIT injuries and eight with associated intraabdominal injuries. There were 24 injuries in the 21 patients consisting of 15 perforations, five contusions, two seromuscular tears, and two gangrene from mesenteric injury. The small intestine was involved in 11 patients, colon six, stomach five, duodenum one and rectum one. Seven (35%) patients had associated extraabdominal injuries. Treatment consisted of simple closure of perforations, over sewing of contusions, resection and anastomosis for gangrene and repair with protective stoma for the rectal injury. One patient each developed prolonged ileus, urinary tract infection and chest infection, respectively postoperatively. Mortality was 28%, all of who had associated intraabdominal or extraabdominal injuries. CONCLUSION: Gastrointestinal injury from blunt abdominal trauma in children, though uncommon, carries a high mortality, usually from associated intraabdominal or extraabdominal injuries. PMID- 15884286 TI - Hepatitis C virus seroprevalence among mothers delivering at the Korle-Bu Teaching Hospital, Ghana. AB - OBJECTIVES: To determine the Hepatitis C virus (HCV) carrier rate among mothers, and to determine if selected sociodemographic characteristics are associated with HCV seropositivity. DESIGN: Maternity Unit of the Korle-Bu Teaching Hospital (KBTH). SETTING: A cross-sectional serological survey of mothers delivering at the KBTH. METHODS: Women who had singleton live births and fresh stillbirths in the two labour wards of the KBTH were randomly selected for screening from 1st March to 30th September, 2001. A structured pre-tested questionnaire was used by trained research assistants to collect and record data on medical and sociodemographic characteristics of the subjects. Maternal blood samples were taken and stored at the Public Health Reference Laboratory. The second generation Murex diagnostics ELISA kit was used to test the maternal sera for HCV antibodies. RESULTS: Sixteen (2.5%, 95% CI, 1.5-4.1%) of the 638 subjects were HCV seropositive. None of the medical and sociodemographic characteristics examined showed any association with HCV seropositivity. No subject or her sexual partner was a drug injector. CONCLUSION: The carrier rate of 2.5% of HCV infection found in this study is near the top end of the range found in unselected pregnant populations from other parts of the world. Knowing the prevalence rate of HCV infection in our pregnant population will help policy makers on the cost effectiveness of available intervention measures. PMID- 15884287 TI - Causes and clinical characteristics of chronic cor-pulmonale in Ethiopia. AB - OBJECTIVE: To determine the clinical characteristics and underlying causes of chronic cor-pulmonale in Ethiopian patients. DESIGN: A hospital based cross sectional study. SETTING: Tikur Anbessa Hospital, a large referral hospital in Addis Ababa, Ethiopia. SUBJECTS: Fourty two consecutive patients referred to the chest clinic of Tikur Anbessa Hospital, Addis Ababa, Ethiopia. RESULTS: Bronchial asthma (36%), chronic fibrocavitary tuberculosis (31%) and chronic bronchitis/emphysema (33%) were the most frequent underlying causes and occurred either singly or in combination in 36(86%) of patients. Interstitial lung disease occurred in five (11.9%) patients. Right-sided heart failure and cyanosis were the most common clinical presentations. Secondary polycythemia was noted in 32 of 40 patients (80%). The role of pulmonary vascular diseases including pulmonary schistosomiasis as a cause of pulmonary heart disease could not be ascertained but appeared to be insignificant. CONCLUSION: Chronic persistent asthma is a frequent underlying cause of chronic obstructive pulmonary disease and chronic cor-pulmonale compared to smoking related chronic bronchitis/emphysema in Ethiopia. The preventive strategy of chronic cor-pulmonale includes optimal treatment of bronchial asthma, early diagnosis and effective treatment of tuberculosis and health education to avoid cigarette smoking. PMID- 15884288 TI - Profile of suicide in Dar es Salaam. AB - OBJECTIVE: To investigate individual socio-demographic characteristics of suicides, the reasons, methods and means employed to commit suicide. DESIGN: A descriptive prospective study of suicides. A structured questionnaire was employed to enquire the details of the itemised objectives. SETTING: Muhimbili National Hospital--in urban Dar es Salaam. SUBJECTS: Fifty three males and 47 females consecutive suicides aged 15 to 59 years RESULTS: The mean age for suicides was found to be 28.2 years. Males were more than females and were ten years older. Sixty two percent of the subjects were single, 30% married. Seventy two percent had primary school education, 19% secondary education. Main reasons for committing suicide were established in 61 cases of which 57.3% (35/61) were due to severe marital and family conflicts, overwhelming disappointments in love affairs and unwanted pregnancies. Eleven subjects with chronic somatic illnesses killed themselves due to unbearable physical pain and overwhelming economic deprivations motivated ten subjects to take their lives. Sixty nine subjects poisoned themselves predominantly using anti-malarials and pesticides while 27 hanged themselves. A third of the suicides consumed alcohol frequently and a quarter of the suicides were HIV positive, a rate twice the national prevalence for sexually active adults. CONCLUSION: Comparatively, women became vulnerable to suicide at a younger age. Dysfunctional social networks played a predominant role among suicides. Family and marital conflicts need closer social attention and timely counseling. Patients with chronic medical conditions and frequent alcohol use need effective exploration concerning suicidal ideation to avert self annihilation. A policy to control prescriptions of toxic drugs including pesticides is overdue. PMID- 15884289 TI - Endoscopic variceal band ligation: a local experience. AB - OBJECTIVE: To evaluate the results of endoscopic variceal band ligation (EVBL) in the local set-up. DESIGN: Retrospective analysis of data of all patients who had EVBL. SETTING: Patients having EVBL at the office endoscopy suite. The Nairobi Hospital, the Aga Khan Hospital and M.P. Shah Hospital. METHODS: The varices were diagnosed at oesophagogastroduodenoscopy (OGD) in patients with hemetemesis and EVBL performed after xylocaine throat spray and intravenous pethidine and bendiazepam sedation. Majority of the patients had EVBL performed on an outpatient basis on weekly intervals until variceal obliteration. RESULTS: A total of 43 patients underwent 119 EVBL sessions at which 523 varies were banded. Four patients had active bleeding and the rest had EVBL for secondary prophylaxis. The variceal kill time was shorter and the transfusion requirements were less and none of the patients developed oesophageal strictures. CONCLUSION: Endoscopic variceal band ligation was noted to give better results in the management of patients with oesophagealvarices in the local set-up, when compared to sclerotherapy in the past. PMID- 15884290 TI - Recurrent chylothorax in a patient with non-Hodgkins lymphoma: case report. AB - Spontaneous chylothorax could arise as a complication of Iymphoma. There are no reports on the frequency of it's occurrence. It is associated with a high mortality rate. This is mainly due to severe nutritional deficiencies and wasting. This case describes a patient with non-Hodgkins Iymphoma who developed recurrent bilateral chylothorax requiring repeated pleural aspirations and eventually talc pleurodesis which failed. PMID- 15884291 TI - Death from body packer syndrome: case report. AB - We report a case of death due to the effects of heroin concealed in a woman who was attempting to smuggle the drug into Kenya concealed within her gastro intestinal tract. She was arrested at a Nairobi airport. While under police detention, she expelled a pellet par anum containing the drug then collapsed, after being forcefully fed on a heavy meal. She was taken to hospital in coma where she expelled nine pellets and died three days later. Her blood level of heroin at admission was well beyond the lethal dose. Details are provided of the circumstances leading to her admission into hospital, her stay in hospital and the autopsy findings. At autopsy 88 pellets were retrieved from her body. The main pathological findings included pulmonary and cerebral edema. Despite the presence of free heroin in the gastric contents, toxicological analysis did not demonstrate any heroin or its metabolites in blood or tissue extracts. This case illustrates the challenges in postmortem evaluation of narcotic fatalities and the need to consider all factors such as antemortem history, toxicology results and autopsy findings in forensic diagnosis PMID- 15884292 TI - Behavior of cinnamoyl-isothiocyanate towards carbon, nitrogen and oxygen reagents. AB - Cyclization of cinnamoyl isothiocyanate with nucleophilic reagents either spontaneously or with added a reagents is reported. PMID- 15884293 TI - Synthesis and biological evaluation of novel naphthoquinone derivatives as potential anticancer and antimicrobial agents. AB - Four novel series of 1,4-naphthoquinone derivatives namely 2,3-disubstiuted 4 [(benzothiazol-2-yl)hydrazono]-1,4-dihydronaphthalen-1-one 4a-c, N1-(2,3 disubstituted-4-oxo-1,4-dihydronaphthalen-1-ylidene)-N2-(benzotriazol-1-yl)acetic acid hydrazide 5a-c, 2,3-disubstituted 4-[(5-aryl-2,3-dihyrothiazol-2 ylidene)hydrazono]-1,4-dihydronaphthalen-1-one 7a-c, and 3-methyl-2-substituted carbamoyl or thiocarbamoyl-hydrazinocarbonylmethylthio-1,4-dihydronaphthalene-1,4 dione 9a-c were synthesized. Three of the new compounds were chosen by NCI to be evaluated as anticancer agents and they showed promising activity. All the prepared compounds were tested as antimicrobial agents. PMID- 15884294 TI - Effect of 5-substituted benzylideneaminosalicylic acid on carrageenan-induced ulcerative colitis. AB - Mesalazine, a drug of choice in the management of ulcerative colitis, was chemically modified as 5-(E)-substituted benzylideneaminosalicylic acid by condensing 5-aminosalicylic acid with selected aryl aldehydes with an intention to improve its pharmacological profile. The tests were performed to study their anti-inflammatory effect on carrageenan-induced ulcerative colitis in albino rats. The histopathological findings, serum transaminase level and lipid peroxide content in the intestine and liver were taken as indices of pharmacological activity. The azomethine derivative formed from salicylaldehyde (5-ASASB3) was found to have maximum activity and it reversed the disease symptoms in experimental animals The azomethine derivative markedly reduces the ulcerative colitis when compared with parent molecule, mesalazine. PMID- 15884295 TI - Oral delivery system of insulin microspheres: effect on relative hypoglycemia of diabetic albino rats. AB - The aim of the present work was to investigate the effectiveness of a dosage form approach for monitoring both the inactivation and the absorption process by targeting insulin delivery to the upper region of small intestine. The dosage form is based on the incorporation of insulin with protease inhibitor and absorption enhancer into polyacrylic polymer Eudragit L-100. Insulin microspheres were prepared by solvent evaporation technique. And also study the effect of these microspheres upon the relative hypoglycemia (RH) effect in white diabetic albino rats has been studied in comparison to that produced after subcutaneous injection of bovine insulin solution. The oral administration of formulation with aprotinin and bile salts gave significant (p< 0.01) hypoglycemia when compared with formulation with insulin alone and with insulin and bile salts. However, the duration, course and the intensity of effect were different for each formulation. It was interesting to observe that the co-administration of aprotinin and bile salts produce prolonged and significant reduction of blood glucose level. A reduction of 4.47-36.81% in plasma glucose levels and RH of about 11.7% relative to subcutaneous injection of soluble insulin solution can be achieved by encapsulation along with protease inhibitor and bile salts. PMID- 15884296 TI - RCN calls time on MRSA. PMID- 15884297 TI - A bug's life. AB - This article explains what methicillin-resistant Staphylococcus aureus (MRSA) is, how it is spread and what the real challenges are in healthcare settings in the UK. It explores the different strains of MRSA and points out the main ways to control their spread. It is intended to be a reference source for all nurses. PMID- 15884298 TI - Faith in transplants. AB - The number of people needing organ transplants outstrips supply--and this is particularly acute in black and minority ethnic groups. A reluctance to talk about the possibility of donation after death is coupled with misconceptions about religious teaching on the subject. PMID- 15884299 TI - Who takes the blame? PMID- 15884300 TI - When dignity kills. PMID- 15884301 TI - No place like home. PMID- 15884302 TI - Provision and decontamination of uniforms in the NHS. AB - AIM: To ascertain the provision and decontamination of uniforms within a cross section of NHS trusts in the UK and to compare policies regarding their use. METHOD: A questionnaire was circulated to 170 NHS trust infection control teams in the UK. Eighty-six (51 per cent) responses were received, which represented 101 NHS trusts. RESULTS: Less than half of the trusts (47 per cent) provide adequate numbers of uniforms to allow a clean uniform per shift. Only 26 per cent had adequate on-site staff changing facilities and 65 per cent did not launder uniforms. The majority of nursing staff (91 per cent) were compelled, by a combination of these factors, to launder their uniforms at home. Few were provided with any guidance on how to do this safely. CONCLUSION: There is an urgent need for minimum standards to be set for the provision of uniforms, laundering and changing facilities, to minimise the potential for spread of healthcare-associated infections. PMID- 15884303 TI - Implementing moving and handling in higher education. AB - Moving and handling education for staff and students was implemented by one higher education institution and examined across its pre-registration health programmes. This article describes related policy and is of relevance to nurse educators involved in moving and handling, back care advisers and mentors of students in the clinical environment. PMID- 15884304 TI - Supporting smoking cessation and dietary change. AB - This article, the fourth of eight in the health promotion series, aims to help nurses support clients make changes in their smoking and dietary behaviours. PMID- 15884305 TI - The Glasgow Coma Scale and other neurological observations. AB - The primary tool used by nurses to assess a patient's neurological status is the neurological observation chart incorporating the Glasgow Coma Scale. This article explains the correct use of the chart and how to interpret the findings. PMID- 15884306 TI - Palliative care. PMID- 15884307 TI - Having it all. PMID- 15884308 TI - Balancing act that works. PMID- 15884309 TI - Fighting back. PMID- 15884310 TI - Scope of practice: a matter of skills, knowledge and erudition. PMID- 15884311 TI - Orofacial pain: how much is it a local phenomenon? PMID- 15884312 TI - Products and publishing. PMID- 15884313 TI - Wine and teeth. PMID- 15884314 TI - The role of parafunctions, emotions and stress in predicting facial pain. AB - BACKGROUND: The authors conducted a study to examine the degree to which parafunctions and emotional states predicted jaw pain in subjects with temporomandibular disorder (TMD) and control subjects. METHODS: Ninety-six subjects diagnosed with myofascial pain, myofascial pain and arthralgia, disk displacement or no TMD symptoms participated. The authors used experience sampling methodology to collect data on pain, behaviors and emotions. They paged subjects approximately every two hours, but not during sleep. When paged, subjects completed a brief questionnaire containing rating scales of jaw pain, masticatory muscle tension, time and intensity of tooth contact, mood and stress level. RESULTS: Analyses of variance showed that groups differed significantly (P < .05) in terms of pain; masticatory muscle tension; and a composite variable measuring time and intensity of contact; mood; and stress. The two myofascial pain groups scored higher on these measures than did the group with disk displacement and the control group. The authors used masticatory muscle tension, the composite variable, mood and stress to predict jaw pain using linear regression. The model was significant and accounted for 69 percent of the variance in jaw pain. Because tension was so highly correlated with jaw pain, the authors removed this variable and re-ran the analysis. The second model also was significant and accounted for 46 percent of the variance in jaw pain. CONCLUSIONS: Parafunctional behaviors, especially those that increase muscle tension, and emotional states are good predictors of jaw pain levels in patients with TMD and healthy control subjects. CLINICAL IMPLICATIONS: Treatment that helps patients reduce parafunctions, excess masticatory muscle tension, stress and emotional distress should be effective in reducing TMD pain. PMID- 15884315 TI - Are female patients with orofacial pain medically compromised? AB - BACKGROUND: Chronic pain has been linked to various medical conditions. The authors assessed whether certain medical conditions are more prevalent in female patients with orofacial pain than in age-matched controls. METHODS: The study sample consisted of 87 female adult subjects from a university-based orofacial pain center (OPC) and 87 age-matched female subjects from a university-based undergraduate dental clinic (UDC). Subjects were evaluated between February 2003 and July 2003, and they completed a standardized, 78-question medical history questionnaire as part of routine clinical protocol. Using the subjects' medical histories, the authors compared 11 major medical categories and 77 individual conditions for both groups. For statistical analyses, the authors used nonparametric Kolmogorov-Smirnov Z tests and chi2 tests and calculated odds ratios (ORs). RESULTS: The medical histories of subjects in the OPC group indicated a significantly greater number of medical conditions than did those of the subjects in the UDC group (Z = -4.411, P < .0001). Specifically, subjects in the OPC group reported having significantly more neurological (Z = -5.304, P < .0001), gastrointestinal (Z = -2.897, P = .004), pulmonary (Z = -2.298, P = .022), dermatologic (Z = -2.984, P = .003) and other conditions (Z = -2.885, P = .004) than did subjects in the UDC group. Subjects in the OPC group reported having 12 individual medical conditions significantly more often (P < .05, ORs ranged from 2.5 to 9.7) than did subjects in the UDC group. CONCLUSIONS: Female patients with orofacial pain complaints appear to have more systemic problems than do female patients seeking routine dental care. CLINICAL IMPLICATIONS: The presence of multiple medical conditions can influence orofacial pain management options and treatment outcomes. Patients with more medically complicated orofacial pain may require treatment on a multidisciplinary basis. PMID- 15884316 TI - Trigeminal neuralgia in a patient with multiple sclerosis and chronic inflammatory demyelinating polyneuropathy. AB - BACKGROUND: Trigeminal neuralgia (TN) is characterized by unilateral, severe, brief, stabbing, recurrent pain in the distribution of one or more branches of the fifth cranial nerve. Symptomatic or secondary TN involves TN-like pain that develops owing to a central nervous system lesion (benign or malignant) or to multiple sclerosis (MS). CASE DESCRIPTION: The authors present a report of a unique case of a 43-year-old patient with unilateral TN, MS and concomitant chronic inflammatory demyelinating polyneuropathy. The facial pain preceded any other manifestations of the systemic disorders, and only after repeated neurological examinations were these diagnoses established. CLINICAL IMPLICATIONS: Magnetic resonance imaging of the brain and repeated neurological evaluations should be implemented in all patients with TN to rule out the presence of underlying disease. The dental practitioner should be familiar with TN to avoid unnecessary dental interventions and ensure prompt initiation of appropriate treatment. PMID- 15884317 TI - Microtensile bond strength of a total-etching versus self-etching adhesive to caries-affected and intact dentin in primary teeth. AB - BACKGROUND: The objec tives of this study were to determine microtensile bond strengths of two dentin adhesives and to compare the micromorphological structure of the resin/dentin interface in caries-affected dentin with that of intact dentin. METHODS: The authors randomly divided 40 proximal dentinal carious primary teeth and 40 noncarious anterior primary teeth into two groups (self etching and total-etching). They used a caries-detecting dye as an indicator of the need to remove the outer carious dentin. The authors restored the teeth with a hybrid resin-based composite. After 24 hours' storage in 37 C water, specimens were sectioned and shaped to form a curved section with a cross-sectional area of 1 square millimeter, then tension was applied until they fractured. The authors prepared the resin/dentin interfaces for the two bonding systems and examined them in 10 occlusal carious and 10 noncarious teeth. STATISTICAL ANALYSIS: The bond strengths for intact and caries-affected dentin within the same group were analyzed via a t test. The authors compared the remaining dentin thickness (RDT) and dentin hardness using analysis of variance and the least significant difference test at the .05 level of significance. RESULTS: The self-etching adhesive demonstrated no statistical difference in bond strength between intact and caries-affected dentin. However, the total-etching adhesive demonstrated different bond strengths for intact and caries-affected dentin. Moreover, the RDT of specimens with intact and caries-affected dentin was not significantly different, whereas the dentin hardness of caries-affected dentin was significantly lower than that of intact dentin. The authors found a thicker hybrid layer in intact and caries-affected dentin of specimens in the total etching group. CONCLUSION: The adhesives exhibited significantly different bond strengths in intact dentin of primary teeth. However, they exhibited similar bond strengths in caries-affected dentin. PMID- 15884318 TI - Malocclusion associated with osteocartilaginous loose bodies of the temporomandibular joint. AB - BACKGROUND: The authors review the literature regarding osteocartilaginous loose bodies (that is, secondary synovial chondrometaplasia or secondary synovial chondromatosis) in the temporomandibular joint (TMJ), present a case report and stress the importance of early diagnosis. CASE DESCRIPTION: A 57-year-old woman was referred to an orthodontist with a chief complaint of bite changes that took place over several years as the patient intermittently experienced TMJ problems. The authors noted radiopacities around the right TMJ space on a panoramic radiograph. They referred the patient to an oral and maxillofacial surgeon for treatment. CLINICAL IMPLICATIONS: Asymmetrical occlusal changes in a nongrowing adult with progressive shifts from Class I to Class III malocclusion unilaterally may indicate a space-occupying lesion in the TMJ space on the affected side. PMID- 15884319 TI - Treatment of a mucogingival defect associated with intraoral piercing. AB - BACKGROUND: In the previous decade, a notable increase in body adornment through tattooing and piercing has been a cause for concern among both dental and medical professionals. The author reports on the clinical consequences of wearing oral jewelry, specifically periodontal injury that requires surgical intervention. She also presents a general literature review of dental and medical consequences of wearing oral jewelry. CASE DESCRIPTION: A 20-year-old woman with a tongue piercing had severe periodontal recession in lingual aspect of the mandibular incisal area proximal to the location of the oral jewelry. The author used a connective tissue graft to correct the defect via root coverage and an addition to the gingival width. CLINICAL IMPLICATIONS: Wearing intraoral jewelry can lead to the development of severe mucogingival defects and necessitates careful and comprehensive periodontal evaluations on a regular basis to monitor attachment loss and damage to dental structures. Patients must be educated about these risks through counseling, patient information brochures and individual case documentation. PMID- 15884320 TI - The major part of dentistry you may be neglecting. AB - Observation of occlusion, providing patient education about occlusion and treatment of occlusal conditions sadly are neglected in the profession. Occlusal equilibration is one of the major treatments for occlusally oriented diseases, and I estimate that this procedure is not accomplished frequently by many practitioners. I have discussed the conditions needing occlusal equilibration and suggested procedures for the conditions. I encourage practitioners needing education in occlusion to seek it. PMID- 15884321 TI - Occupation-related allergies in dentistry. AB - BACKGROUND: Allergies to natural rubber latex (NRL) were unknown in dentistry until 1987. That changed with the publication of a report documenting NRL-based anaphylaxis in a dental worker. This case and others prompted regulatory and manufacturing changes in rubber products and increased awareness throughout the profession. However, other common dental chemicals cause allergic reactions and irritation and often are handled with insufficient precautions. Although recognition of NRL allergy has improved, awareness of other potential allergens and irritants in dentistry still is limited. OVERVIEW: Recent research indicates that the prevalence of NRL protein allergy may be decreasing. In contrast, occupation-related dermatoses associated with other dental products may be more common. Encounters with bonding agents, disinfectants, rubber, metals and detergents can cause occupation-based irritant contact dermatitis and allergic contact dermatitis. These conditions may be found in more than one-quarter of dental and medical personnel. Therefore, dental-specific information about the recognition and management of allergic and irritant reactions is needed. CONCLUSIONS AND CLINICAL IMPLICATIONS: The prevalence of occupation-related dermatitis may be increasing in dentistry. Reducing exposure to potential irritants and allergens and educating personnel about proper skin care are essential to reversing this trend. PMID- 15884322 TI - The challenge of esthetic dentistry and elective services. AB - Many dentists would like to increase the number of esthetic and elective procedures they perform. What may be holding them back is the ability to communicate with, educate and motivate patients in the area of esthetic and elective dentistry. While implants, esthetic dentistry or elective dentistry is not for every patient, each of these areas has improved the quality of many people's lives, allowing the dental profession to continue to serve the public in the broadest possible way. PMID- 15884323 TI - Dentists' participation and children's use of services in the Indiana dental Medicaid program and SCHIP: assessing the impact of increased fees and administrative changes. AB - BACKGROUND: The authors conducted a study to evaluate whether administrative changes, including higher fee schedules for dental services in the Indiana dental Medicaid program and the State Children's Health Insurance Program (SCHIP), were associated with improved dentist participation and utilization of dental services by children. METHODS: The authors evaluated dentists' participation and children's use of services for the two years before fees were increased to 100 percent of the 75th percentile of usual and customary fees, compared with two years after the increase. They obtained administrative data from the Indiana Department of Family and Social Services Administration and the Indiana Department of Public Health to determine participation rates and service use. RESULTS: The number of dentists seeing a Medicaid-enrolled child increased from 770 in fiscal year (FY) 1997 to 1,096 in FY 2000. The number of Medicaid-enrolled children with any dental visit increased from 68,717 (18 percent) to 147,878 (32 percent), with little difference between children enrolled through the Medicaid SCHIP and traditional Medicaid programs by FY 2000. The mean number of visits per child per year and the mean number of procedures per child per year remained relatively constant. The cost per enrolled child increased from dollars 1.70 to dollars 6.70 per month, while the cost per child with a visit increased from dollars 9 to dollars 21 per month. CONCLUSION: The increase in fees and changes in administration of the Indiana dental Medicaid program were positively associated with improved dentist participation and children's use of dental services. PRACTICE IMPLICATIONS: Changes beyond increasing fees to 100 percent of the 75th percentile may be needed if Medicaid-enrolled children are to have access to dental care commensurate with their lower oral health status and greater need for services. Sustained fee increases also are important. As of 2003, no increase in dental fees had occurred in the Indiana Medicaid program since the increase in FY 1998. PMID- 15884324 TI - Dental visits and access to dental care among Maryland schoolchildren. AB - BACKGROUND: Regular dental visits afford an opportunity for dentists to provide preventive services and to diagnose and treat disease. Not all children, however, have equal access to these services. METHODS: The authors conducted this study to describe access to and utilization of oral health care services for Maryland schoolchildren in kindergarten and third grade. They obtained data from a questionnaire filled out by parents or guardians participating in the Survey of the Oral Health Status of Maryland School Children, 2000-2001 (N = 2,642). Outcome variables included having a dental visit in the last year, prophylaxis in the last year, usual source of medical care and usual source of dental care. Descriptor variables included region, grade, race/ethnicity, eligibility for free or reduced-fee meals, parents' or guardians' education and dental insurance status. RESULTS: Overall, general dental visit and dental prophylaxis visit rates were similar (74.1 and 71.3 percent, respectively). Schoolchildren, however, were more likely to have had a usual source of medical care than of dental care (96.0 and 82.9 percent, respectively). Third graders, those ineligible for free or reduced-fee meals and those with some dental insurance coverage were more likely to have received a prophylaxis in the last year and were more likely to have a usual source of dental care. Non-Hispanic white and non-Hispanic black schoolchildren also were more likely to have had a usual source of dental care than were Hispanics. CONCLUSIONS: Schoolchildren most likely to have received regular preventive dental care were those who had parents or guardians with financial resources. Medicaid and State Children's Health Insurance Program (SCHIP) provide safety nets, but these programs could be improved. PRACTICE IMPLICATIONS: Dentistry's challenge is to determine which characteristics are unique to those who visit the dentist regularly and use this information to help meet the needs of the underserved. PMID- 15884326 TI - For the dental patient... Do you grind your teeth? PMID- 15884325 TI - Going the defamation route: with unlawful dismissal not an option, ousted employee sues for defamation and prevails. PMID- 15884327 TI - Controversial agricultural air monitoring program gets under way. PMID- 15884328 TI - U.S. companies get nervous about EU's REACH. PMID- 15884329 TI - An introduction to the National Environmental Methods Index. PMID- 15884330 TI - Characterization of trophic transfer for polychlorinated dibenzo-p-dioxins, dibenzofurans, non- and mono-ortho polychlorinated biphenyls in the marine food web of Bohai Bay, North China. AB - Many investigations have highlighted the bioaccumulation of dioxins in animals, but little is known about the trophodynamics of dioxins in the food web. In this study, the trophic transfer of nine dibenzo-p-dioxin (PCDD) congeners, eleven dibenzofuran (PCDF) congeners, and twelve non-, mono-ortho polychlorinated biphenyl (non- and mono-ortho PCBs) congeners in a marine food web were determined. The concentrations of PCDDs, PCDFs, non- and mono-ortho PCBs were analyzed in phytoplankton/ seston, zooplankton, three invertebrate species, six fish species, and one seabirds species collected from Bohai Bay, representing approximately 4 trophic levels based on stable nitrogen isotope values. Positive relationships were found between trophic levels and lipid equivalent concentrations of non- and mono-ortho PCBs except for PCB-77, PCB-81, PCB-126, PCB-156, and PCB-167, indicating bioaccumulation of these compounds in this food web. But lipid equivalent concentrations of low chlorinated 2,3,7,8-substituted PCDD/Fs did not exhibit statistically significant trends with trophic levels. And lipid equivalent concentrations of high chlorinated 2,3,7,8-substituted-PCDD/Fs and three non-2,3,7,8-substituted-PCDD/Fs declined significantly with increasing trophic levels providing that these isomers undergo trophic dilution. The similarity in log Kow values for non-, mono-ortho PCBs, non-2,3,7,8-substituted PCDD/Fs, and some 2,3,7,8-substituted-PCDD/Fs suggests that the difference of trophic transfer is mainly due to their different metabolic transformation rates. PMID- 15884331 TI - Wet deposition of persistent organic pollutants to the global oceans. AB - Wet deposition fluxes of polychlorinated biphenyls and polychlorinated dibenzo-p dioxins and furans to the Atlantic Ocean have been estimated by combining meteorological satellite data and measured atmospheric field concentrations. They are then compared to other atmospheric depositional mechanisms on a global scale. Additional features not treated in traditional studies are addressed such as contaminant adsorption onto raindrops and enhancement of dry gaseous diffusive fluxes due to rain-induced turbulence. Wet deposition estimates show a high spatial and seasonal variability, with maxima located in the Intertropical Convergence Zone (ITCZ) and in low-temperature regions. Seasonal variability reflects the northward shift of ITCZ in July. Average wet deposition fluxes estimated for the Atlantic Ocean in this study are 110 and 45 ng m(-2) yr(-1) for sigmaPCB and sigmaPCDD/Fs, respectively. Furthermore, the total wet deposition to the Atlantic results in 4100 kg yr(-1) (sigmaPCB) and 2500 kg yr(-1) (sigmaPCDD/Fs). Model validation shows good agreement with available coastal data measurements of wet deposition fluxes. When compared to other atmospheric depositional mechanisms and during precipitation events, wet deposition is found to be dominant. However, when raining events and non-raining time periods are integrated, air-water diffusive exchange fluxes acquire an important role, which can be dominant in some regions and for some POPs. PMID- 15884332 TI - Time trends of atmospheric PBDEs inferred from archived U.K. herbage. AB - Aerial portions of vegetation receive the bulk of their burden of persistent organic pollutants (POPs) from the atmosphere. Vegetation can therefore be a useful indicator of the changing atmospheric burden of POPs. Samples of archived pasture, collected from Rothamsted Experimental Station in the United Kingdom between 1930 and 2004, were analyzed for a range of polybrominated diphenyl ethers (PBDEs). PBDEs could not be routinely detected in the pre-1970 samples. Thereafter, the dominant congeners BDE 28, 47, 49, 99, 100, 153, 154, and 183 were frequently detected. The general trend was (a) a rise through the 1970s; (b) a minipeak in the mid-1980s, strongly influenced by one particularly high sample for 1984; (c) values remaining high through the late 1980s/1990s; (d) an indication of a more recent decline for all congeners (except BDE-28), consistent with recent restrictions on PBDE usage in Europe. These trends were compared to recent modeled estimates of U.K. PBDE emissions. The congener profiles of technical mixtures, U.K. air, soil, and pasture were compared and shown to be broadly similar. The implications for environmental release mechanisms are discussed. PMID- 15884333 TI - Corrosion of unexploded ordnance in soil-field results. AB - Unexploded ordnance (UXO) are found on 400 000 Ha of land across 1400 different sites in the United States. In many cases, UXO contain high explosives posing a risk to groundwater quality. This paper provides results from a field survey of 14 inactive Army ranges distributed throughout the mainland United States. Metal samples and adjacent soil were collected from 161 UXO. Soil samples were analyzed for a variety of chemical and physical properties. Metal samples were analyzed for pitting corrosion. Climate data were also compiled. Of these analyses, only rainfall correlated with extent of corrosion. Samples had been buried from the Civil War era through the 1990s. Photographs taken in the field illustrate that corrosion occurs more rapidly on the underside of a UXO. Field observations also revealed that low-order detonations and UXO are difficult to distinguish in the field. The deepest individual pit measured on any of the 161 metal samples was 2400 microm. Low rainfall environments had shallower average pit depths (590+/ 130 microm)than moderate and high rainfall environments, which produced deeper average pit depths (940+/-90 microm). The UXO studied in this effort were not a significant source of explosives to the unsaturated soil environment. PMID- 15884334 TI - Synergic effect of gold mining and damming on mercury contamination in fish. AB - Since the late 1980s, several studies have shown that human populations in the Amazon basin are exposed to high mercury levels in their fish diet. Gold mining, which releases the metal during the amalgamation process and erodes soils naturally rich in mercury, is regarded as the main contamination source. Here, we present the results of a comparative study of mercury distribution in the water and fish of two adjacent rivers in French Guiana, with and without gold mining activities. As a consequence of a marked difference in suspended particulate matter between the two systems, total mercury concentrations in unfiltered water samples were higher in the mined river (25.4-34.9 ng L(-1)) as compared to the reference one (2.1-5.4 ng L(-1)). Surprisingly, no significant differences were observed in mercury concentrations between 13 common fish species at upstream sites. In sharp contrast, mercury concentration of fish caught downstream a hydroelectric reservoir, where the two rivers flow, was up to 8-fold higher than that upstream. Mercury speciation measurements allowed one to relate these differences in fish to the water distribution of monomethylmercury, the mercury chemical species that biomagnifies along aquatic foodwebs. Indeed, mean dissolved monomethylmercury concentrations were low and similar in both rivers (0.03-0.06 ng L(-1)), while they were 10 times higher (up to 0.56 ng L(-1)) in the water outflowing the hydroelectric dam. Dissolved monomethylmercury determinations along a water column profile suggest that methylation of inorganic mercury occurs in the deep anoxic part in reservoir. We conclude that mercury mobilization related to gold mining is not solely sufficient to account for high concentrations in fish and that environmental conditions that favor mercury methylation, such as anoxia, are needed. PMID- 15884335 TI - Analysis of oxidative DNA damage 8-hydroxy-2'-deoxyguanosine as a biomarker of exposures to persistent pollutants for marine mammals. AB - An oxidative DNA biomarker, 8-hydroxy-2'-deoxyguanosine (8-OHdG), was determined in the livers and kidneys of stranded or by-caught cetaceans along the Taiwan coast through isotope-dilution liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) to evaluate the feasibility of analyzing the DNA adduct in marine mammals and then to study the association between 8-OHdG and levels of polychlorinated biphenyls (PCBs) and dichlorodiphenyldichloroethylene (DDE) in the blubbers of the cetaceans. The mean values of the 8-OHdG from the liver and kidney samples were 19.83+/-10.00 pmol/micromol deoxyguanosine (dG) (6.90-53.53 pmol/micromol dG) and 19.16+/-7.48 pmol/micromol dG (5.36-39.36 pmol/micromol dG), respectively. In general, 8-OHdG was not related to the general health status of the by-caught and stranded animals and also was not related to species. However, the levels of 8-OHdG had a positive correlation with concentrations of PCBs, but not DDE, in female cetacean livers. In addition, when selected coplanar PCBs (dioxin-like congeners) were used to compare the 2,3,7,8-tetrachlorodibenzo p-dioxin equivalents (TEQs) with 8-OHdG of by-caught cetaceans, a high positive correlation (r = 0.80, p < 0.01) was found in mature female animals. Thus, the detection of 8-OHdG in marine mammals with isotope-dilution LC/MS/ MS is possible, and the study of the relationship between oxidative DNA damage and environmental contaminants under natural exposure indicates that the level of 8 OHdG in female cetacean livers is associated with coplanar PCBs and the factor of sexual maturity. PMID- 15884336 TI - Mixtures of estrogenic contaminants in bile of fish exposed to wastewater treatment works effluents. AB - Most effluents from wastewater treatment works (WwTWs) contain estrogenic chemicals that include steroidal estrogens and xenoestrogens. We investigated the nature of mixtures of estrogenic contaminants taken up by two species of fish exposed to two WwTWs effluents. Sexually immature rainbow trout, Oncorhynchus mykiss, and sexually mature roach, Rutilus rutilus, were exposed to tap water, river water, or one of two estrogenic WwTWs effluents for up to 10 days, when the fish were sacrificed and tissues removed for chemical analysis. Estrogenic contaminants in the bile and gonads were hydrolyzed, concentrated by solid-phase extraction, and fractionated by RP-HPLC. Active fractions were detected and quantified using a yeast estrogen receptor transcription screen (YES assay) and the identities of estrogenic components in the fractions determined by GC-MS. Bile from rainbow trout exposed to either tap water or river water contained low amounts of 17beta-estradiol (E2) and estrone (E1) with a total estrogenic activity (mean+/-standard error) of 10+/-5 and 31+/-9 ng of E2 equivalents/mL (ng of E2eq/mL) for male and female fish, respectively. In effluent-exposed trout the total estrogen content of bile was considerably higher with the following composition and concentrations (ng of E2eq/mL) of individual estrogens: E2 (male, 591+/-125; female, 710+/-207), E1 (male, 338+/-75; female, 469+/-164), ethinylestradiol, EE2 (male, 32+/-2; female, 40+/-6), nonylphenol (NP) and short chain NP polyethoxylates (male, 21+/-4; female, 22+/-3). An additional estrogenic compound, 17beta-dihydroequilenin (DHQ), was identified for the first time in effluent-exposed fish, and was present in trout bile at concentrations of (male) 40+/-9 and (female) 30+/-5 ng of E2 eq/mL. DHQ, E2, E1, and EE2, but not NP or NP polyethoxylates, were also detected in bile of effluent-exposed roach, and the concentrations of all these steroidal estrogens in ng of E2eq/mL were lower in male (E2, 62+/-2; E1, 35+/-11; EE2, 10+/-2; DHQ, 1+/-1) compared with female (E2, 740+/-197; E1, 197+/-37; EE2, 40+/-6; DHQ, 8+/-2) roach. The synthetic estrogen EE2 was also detected in the testes and ovaries of effluent-exposed roach. This study shows that a mixture of estrogenic contaminants present in WwTWs effluents bioconcentrate in fish tissues, resulting in the induction of vitellogenin, and are likely to contribute to feminizing effects in wild fish living in U.K. rivers. The composition of the mixture of estrogenic contaminants in the bile is species dependent and may determine the susceptibility of fish to the effects of exposure to estrogenic effluents. PMID- 15884337 TI - Novel flame retardants, 1,2-bis(2,4,6-tribromophenoxy)ethane and 2,3,4,5,6 pentabromoethylbenzene, in United States' environmental samples. AB - Two brominated flame retardants, 1,2-bis(2,4,6-tribromophenoxy)ethane (TBE) and 2,3,4,5,6-pentabromoethylbenzene (PEB), were detected and identified in ambient air samples from various sites in the United States. The identifications were confirmed by comparing the gas chromatographic retention times and mass spectra of the compounds found in the environment with those of authentic materials. Generally, the TBE concentrations in air were comparable to those of tetra- through hexabrominated diphenyl ethers (PBDEs) and often higher than those of decabromodiphenyl ether (BDE-209). The atmospheric TBE concentrations at locations in the southern United States were higher than those in the northern United States. TBE was also found in a sediment core from Lake Michigan; the concentrations of TBE increased with time, were lower than those of BDE-209, but were approximately 10 times higher than the sum of BDE-47, -99, and -100. The maximum PEB concentration in Chicago air was 550 pg/ m3, which was 10 times higherthan the concentration of total PBDEs in this sample. In general, the concentrations of PEB in air samples were low but detectable and were less than those of PBDEs. PEB was not found in the sediment core from Lake Michigan. These occurrences of relatively high concentrations of TBE and PEB in environmental samples may reflect the increasing usage of these compounds as flame retardants. PMID- 15884338 TI - Characterzation of colloidal and humic-bound Ni and U in the "dissolved" fraction of contaminated sediment extracts. AB - The dissolved phase of environmental aqueous samples is generally defined by filtration at 0.2 microm or even 0.45 microm. However, it is also acknowledged that colloids <0.2 microm suspended in the aqueous phase can be important for determining contaminant availability and mobility. We have used flow field-flow fractionation (FI FFF) and size exclusion chromatography (SEC) coupled to UV absorbance (UVA) and inductively coupled plasma mass spectrometry (ICP-MS) to study the dissolved organic matter (DOM) and colloidal binding of U and Ni in water extracts of sediments collected from a contaminated area of the Savannah River Site, a U.S. Department of Energy former nuclear materials production and processing facility, near Aiken, SC. High-performance SEC-UVA-ICP-MS was well suited to the separation of DOM overthe molecular weight (MW) range of approximately 200-7000 Da. The ICP-MS element specific data indicated that a significant fraction of U was associated with DOM. Uranium exhibited a bimodal distribution and the other fraction was greater than the exclusion limit for the column and coeluted with Al. Flow FFF was used to size this fraction as colloidal with an approximate effective spherical diameter of 0.09-0.12 microm. Element specific ICP-MS data confirmed that U and Al were associated with the colloidal phase. High-field FI FFF was also applicable to sizing DOM but resolution was poorer than SEC. The results of this study suggest that "dissolved" U at this site is predominantly either complexed by DOM or bound to a colloidal fraction while Ni is predominately present as labile complexes or the free cation and, therefore, potentially bioavailable. PMID- 15884339 TI - Boron isotopes in the Seine River, France: a probe of anthropogenic contamination. AB - Boron concentrations and isotopic compositions have been measured in the dissolved load of the Seine Basin rivers, France. Hydrology and chemistry of the Seine River and its tributaries are strongly influenced by human activities, as the anthropogenic pressure on the Seine catchment is one of the highest in Europe. The samples were collected between 1994 and 1996 during various stages of flow, complemented by a time-series of the Seine River in Paris for 1 yr. In particular, the decennial flood event of winter 1994 was sampled. Boron appears to be conservative in rivers and not influenced by adsorption onto suspended matter and/or consumption by microorganisms. Despite the complexity of the Seine River system, dissolved boron and its isotopes are found to be suitable tracers of contamination. The total dissolved boron of the Seine River at Paris is explained by the contribution from three distinct components: Urban effluents constitute 65% of the boron discharge measured in the Seine River whereas agriculture-affected waters contribute less than 10% with a more marked influence during high water discharges. Rainwater contribution is important (25% mean), reaching 30% of dissolved boron during high flood events. PMID- 15884340 TI - Organochlorine pesticides in agricultural soil and vegetables from Tianjin, China. AB - Samples of eight types of vegetables, the rhizosphere soils, and bulk soils were collected from two sites (A and B) in Tianjin, China for the determination of hexachlorocyclohexane isomers (HCHs) and dichlorodiphenyltrichloroethane and metabolites (DDXs). The average concentrations of total HCHs and DDXs in the bulk soils were 3.6 and 80.1 ng/g for site A and 102 and 235 ng/g for site B, respectively. Relative accumulations of HCHs and DDXs in the rhizosphere soil from site A but not site B were demonstrated. The concentrations of total HCHs and DDXs in vegetable roots were 3.6-60 and 4.2-73 ng/g for site A and 15-152 and 7.1-136 ng/g for site B, respectively. Difference in bioaccumulation among various vegetables, especially between tuber and fibrous vegetables was significant. DDXs in spinach and cauliflower from site B and lindane (gamma-HCH) in cauliflower from both sites and violet from site B exceeded the maximum residual limits. Linear correlation of log-transformed HCHs and DDXs contents between the vegetable roots and the rhizosphere soils suggests the direct uptake of HCHs and DDXs. PMID- 15884341 TI - Detachment-influenced transport of an adhesion-deficient bacterial strain within water-reactive porous media. AB - Bacteria and carboxylate-modified microsphere transport experiments were performed in glass bead packed columns in order to examine the distribution of retained colloids on the sediment. Solution pH was allowed to vary from 6.0 to 9.4 across the length of the column (20 cm) in order to examine potential effects of solution chemistry on the retained profiles. Both the microspheres and the bacteria showed retained profiles that deviated strongly from log-linear behavior expected from a spatially invariant colloid deposition rate coefficient. Deviation for the microspheres was in the form of steeper-than-expected decreases in retained concentrations with distance from source. Deviation for the bacteria was in the form of maximum retained concentrations that were located down gradient from the column inlet. Subsidiary experiments with varying elution times showed that detachment during elution moved the peak of mass of retained bacteria down-gradient of the column inlet; however, the disproportional translation of the peak of mass relative to elution time indicated that processes operating during injection produced the initial down-gradient translation of peak concentrations of retained cells. PMID- 15884342 TI - Copper-glyphosate sorption to microcrystalline gibbsite in the presence of soluble Keggin Al13 polymers. AB - Among the most reactive yet largely neglected adsorbents of toxicant species occurring in acidic aquatic environments are the epsilon-Keggin Al13 polyoxocations [AlO4Al12(OH)24(H2O)12(7+)], known generally as Al13 polymers. Here, we report on the sorption of Cu(II), a common ingredient of pesticides, and glyphosate {N-[phosphonomethyl]glycine (PMG)}, a widely applied herbicide, to microcrystalline gibbsite [gamma-Al(OH)3] in the presence of soluble Al13 polymers over the pH range 4-7. In the presence of gibbsite and soluble Al13 polymers, dissolved Cu(II) decreased gradually with pH, achieving a minimum at pH 5.5. Between pH 5.5 and 6.0, however, soluble Cu increased markedly, with approximately 80% of the added metal remaining in solution at pH 5.86. At pH > 6.0, soluble Cu once again decreased, becoming undetectable at pH 7. The anomalous Cu solubilization was attributed to a concomitant deprotonation of soluble Al13 polymers, yielding surface OH groups possessing high affinity for Cu(II). Removal of Cu from solution at pH > 6.0 is facilitated by flocculation of the Al13 polymers to which Cu had sorbed. The sorption behavior of the zwitterionic PMG in the presence of gibbsite and Al13 polymers was consistent with this interpretation, there being a dramatic increase in sorbed PMG at pH > 6.0 as the Al13 polymers deprotonated and flocculated. Copper and PMG loss from solution with increasing pH when both adsorptives were added to the gibbsite-Al13 polymer system was broadly similar to what was observed in the PMG-free systems, although small differences were detected in response to varying the order of adsorptive addition. The inclusion of soluble Al polymers in our experiments exposes a fundamental limitation of models based on but a single inorganic adsorbent as a means to predict the behavior of trace metals and xenobiotic organic compounds in natural systems. PMID- 15884343 TI - Low molecular weight carboxylic acids in oxidizing porphyry copper tailings. AB - The distribution of low molecular weight carboxylic acids (LMWCA) was investigated in pore water profiles from two porphyry copper tailings impoundments in Chile (Piuquenes at La Andina and Cauquenes at El Teniente mine). The objectives of this study were (1) to determine the distribution of LMWCA, which are interpreted to be the metabolic byproducts of the autotroph microbial community in this low organic carbon system, and (2) to infer the potential role of these acids in cycling of Fe and other elements in the tailings impoundments. The speciation and mobility of iron, and potential for the release of H+ via hydrolysis of the ferric iron, are key factors in the formation of acid mine drainage in sulfidic mine wastes. In the low-pH oxidation zone of the Piuquenes tailings, Fe(III) is the dominant iron species and shows high mobility. LMWCA, which occur mainly between the oxidation front down to 300 cm below the tailings surface at both locations (e.g., max concentrations of 0.12 mmol/L formate, 0.17 mmol/L acetate, and 0.01 mmol/L pyruvate at Piuquenes and 0.14 mmol/L formate, 0.14 mmol/L acetate, and 0.006 mmol/L pyruvate at Cauquenes), are observed at the same location as high Fe concentrations (up to 71.2 mmol/L Fe(II) and 16.1 mmol/L Fe(III), respectively). In this zone, secondary Fe(III) hydroxides are depleted. Our data suggest that LMWCA may influence the mobility of iron in two ways. First, complexation of Fe(III), through formation of bidentate Fe(III)-LMWCA complexes (e.g., pyruvate, oxalate), may enhance the dissolution of Fe(III) (oxy)hydroxides or may prevent precipitation of Fe(III) (oxy)hydroxides. Soluble Fe(III) chelate complexes which may be mobilized downward and convert to Fe(II) by Fe(III) reducing bacteria. Second, monodentate LMWCA (e.g., acetate and formate) can be used by iron-reducing bacteria as electron donors (e.g., Acidophilum spp.), with ferric iron as the electron acceptor. These processes may, in part, explain the low abundances of secondary Fe(III) hydroxide precipitates below the oxidation front and the high concentrations of Fe(II) observed in the pore waters of some low-sulfide systems. The reduction of Fe(III) and the subsequent increase of iron mobility and potential acidity transfer (Fe(II) oxidation can result in the release of H+ in an oxic environment) should be taken in account in mine waste management strategies. PMID- 15884344 TI - Quantifying the contribution of different sorption mechanisms for 2,4 dichlorophenoxyacetic acid sorption by several variable-charge soils. AB - Previous research with phenolic, carboxylic, and urea type organic acids demonstrated that hydrophilic sorption was primarily due to anion exchange, which was linearly correlated to chemical acidity (pKa) and the soil anion exchange capacity. However, for dichlorophenoxyacetic acid (2,4-D), sorption by a kaolinitic soil was much higher than expected relative to all other organic acid soil data. The enhanced sorption was hypothesized to involve calcium bridging of 2,4-D to hydrophilic domains. In this study, the mechanisms contributing to 2,4-D sorption by variable-charged soils were probed and quantified by measuring sorption from CaCl2, KCl, CaSO4, KH2PO4, and Ca(H2PO4)2 solutions. Linear sorption coefficients estimated for 2,4-D sorption from the different electrolytes decreased as follows: CaCl2 > KCl > CaSO4 > Ca(H2PO4)2 approximately equal to KH2PO4. Differences in 2,4-D sorption from CaCl2 and phosphate solutions were attributed to sorption by hydrophilic domains, which ranged between 46 and 94% across soils. Differences in 2,4-D sorption from CaCl2 and KCl were attributed specifically to Ca-bridging between 2,4-D's carboxyl group and the silanol edge on kaolinite and quartz and ranged from negligible to 40% depending on the soil mineral type. Differences in sorption from CaCl2 and CaSO4 was attributed to anion exchange, which ranged from 16 to 91%, followed the trends with pKa developed previously for other organic acids, and correlated well to the soil anion to cation exchange capacity ratio (AEC/CEC). The sum of anion exchange and Ca-bridging contributions agreed well with the sorption estimated to be from hydrophilic domains. All other sorption was attributed to hydrophobic processes, which correlated well to a linear free-energy relationship between pH-dependent organic carbon-normalized sorption coefficients and pH-dependent octanol-water partition coefficients developed for several other organic acids. PMID- 15884345 TI - Role for Fe(III) minerals in nitrate-dependent microbial U(IV) oxidation. AB - Microbiological reduction of soluble U(VI) to insoluble U(IV) is a means of preventing the migration of that element in groundwater, but the presence of nitrate in U(IV)-containing sediments leads to U(IV) oxidation and remobilizaton. Nitrite or iron(III) oxyhydroxides may oxidize U(IV) under nitrate-reducing conditions, and we determined the rate and extent of U(IV) oxidation by these compounds. Fe(III) oxidized U(IV) at a greater rate than nitrite (130 and 10 microM U(IV)/day, respectively). In aquifer sediments, Fe(III) may be produced during microbial nitrate reduction by oxidation of Fe(II) with nitrite, or by enzymatic Fe(II) oxidation coupled to nitrate reduction. To determine which of these mechanisms was dominant, we isolated a nitrate-dependent acetate- and Fe(ll)-oxidizing bacterium from a U(VI)- and nitrate-contaminated aquifer. This organism oxidized U(IV) at a greater rate and to a greater extent under acetate oxidizing (where nitrite accumulated to 50 mM)than under Fe(II)-oxidizing conditions. We showthatthe observed differences in rate and extent of U(IV) oxidation are due to mineralogical differences between Fe(III) produced by reaction of Fe(II) with nitrite (amorphous) and Fe(III) produced enzymatically (goethite or lepidocrocite). Our results suggest the mineralogy and surface area of Fe(III) minerals produced under nitrate-reducing conditions affect the rate and extent of U(IV) oxidation. These results may be useful for predicting the stability of U(IV) in aquifers. PMID- 15884346 TI - Arsenate adsorption mechanisms at the allophane-water interface. AB - We investigated arsenate (As(V)) reactivity and surface speciation on amorphous aluminosilicate mineral (synthetic allophane) surfaces using batch adsorption experiments, powder X-ray diffraction (XRD), and X-ray absorption spectroscopy (XAS). The adsorption isotherm experiments indicated that As(V) uptake increased with increasing [As-(V)]0 from 50 to 1000 microM (i.e., Langmuir type adsorption isotherm) and that the total As adsorption slightly decreased with increasing NaCl concentrations from 0.01 to 0.1 M. Arsenate adsorption was initially (0-10 h) rapid followed by a slow continuum uptake, and the adsorption processes reached the steady state after 720 h. X-ray absorption spectroscopic analyses suggest that As(V) predominantly forms bidentate binuclear surface species on aluminum octahedral structures, and these species are stable up to 11 months. Solubility calculations and powder XRD analyses indicate no evidence of crystalline Al-As(V) precipitates in the experimental systems. Overall, macroscopic and spectroscopic evidence suggest that the As(V) adsorption mechanisms at the allophane-water interface are attributable to ligand exchange reactions between As-(V) and surface-coordinated water molecules and hydroxyl and silicate ions. The research findings imply that dissolved tetrahedral oxyanions (e.g., H2PO42- and H2AsO4(2-)) are readily retained on amorphous aluminosilicate minerals in aquifer and soils at near neutral pH. The inner-sphere adsorption mechanisms might be important in controlling dissolved arsenate and phosphate in amorphous aluminosilicate-rich low-temperature geochemical environments. PMID- 15884347 TI - Isotopic constraints on the fate of petroleum residues sequestered in salt marsh sediments. AB - To provide a new perspective on the fate of petroleum in the marine environment, we utilized variations in the natural abundance of radiocarbon (14C) to detect and quantify petroleum residues that have persisted in Wild Harbor sediments, West Falmouth, MA, for more than 30 years. The 5730-yr half-life of 14C makes this isotope ideal for the detection of fossil-fuel-derived contaminants (14C free) within different fractions of natural organic matter (modern 14C content) in environmental matrixes. Samples of both contaminated and uncontaminated sediments were sequentially treated, first by solvent extraction, followed by saponification, and then acid hydrolysis. Radiocarbon analysis of the sediment residues and select extracts was performed to probe for the presence of fossil fuel contaminants and/or their metabolites in different pools of sedimentary organic matter. Our results indicate that the majority of fossil carbon is solvent-extractable and has not been incorporated in the insoluble organic matter in sediment. Unextracted sediments contaminated with petroleum contain significantly less 14C than extracted sediments, and isotope mass balance calculations suggest that up to approximately 9% of the total organic carbon (TOC) in the petroleum contaminated sediment horizons is derived from solvent extractable petroleum. These estimates are similar to values calculated when the total quantities of oil (measured by gas chromatography with flame ionization detector (GC-FID)) are compared to TOC content (determined by elemental analysis). These results pave the way for applications of this isotopic approach to more complex environmental systems where the fate of contaminants is less certain. PMID- 15884348 TI - Determination of microbial carbon sources in petroleum contaminated sediments using molecular 14C analysis. AB - Understanding microbial carbon sources is fundamental to elucidating the role of microbial communities in carbon cycling and in the biodegradation of organic contaminants. Because the majority of anthropogenic contaminants are either directly or indirectly derived from fossil fuels that are devoid of 14C, radiocarbon can be used as a natural inverse tracer of contaminant carbon in the contemporary environment. Here, 14C analysis of individual microbial phospholipid fatty acids (PLFA) was used to characterize the carbon sources utilized bythe active microbial community in salt marsh sediments contaminated by the Florida oil spill of 1969 in Wild Harbor, West Falmouth, MA. A specific goal was to determine whether this community is actively degrading petroleum residues that persist in these sediments. The delta14C values of microbial PLFA in all sediment horizons (contaminated and noncontaminated) matched the delta14C of the total sedimentary organic carbon after petroleum removal, indicating that no measurable metabolism of petroleum residues was occurring. This result agrees with ancillary data such as the delta13C content and distribution of PLFA, and the residual hydrocarbon composition determined by comprehensive two-dimensional gas chromatography (GCxGC) analysis. We hypothesize that microbes have chosen to respire the natural organic matter rather than the residual petroleum hydrocarbons because the former is more labile. Future efforts directed at determining indices of microbial degradation of petroleum hydrocarbons should consider competition with natural organic matter. PMID- 15884349 TI - Vehicle self-pollution intake fraction: children's exposure to school bus emissions. AB - Vehicle self-pollution occurs when a vehicle's emissions migrate to inside that vehicle's passenger compartment. This paper presents values fortwo new parameters: vehicle self-pollution intake fraction (iF(SP)), which is the total fraction of a vehicle's emissions inhaled by all people in the vehicle, and vehicle self-pollution individual intake fraction (iF1SP), which is the fraction of a vehicle's emissions inhaled by an individual in the vehicle. We use results from tracer-gas experiments in California's South Coast Air Basin (SoCAB) to quantify students' iF(SP) and iF1SP for school bus emissions. Six buses were studied during nine runs with windows open and seven runs with windows closed. Reported iF(SP) values (units: per million; min = 10, max = 94, mean = 27) indicate that the total mass of a bus' exhaust inhaled by students commuting on it is comparable in magnitude to the total mass of that bus' exhaust inhaled by all other people in the SoCAB. Reported iF1SP values (units: per million; min = 0.2, max = 2.4, mean = 0.7) indicate that average per capita inhalation of emissions from any single bus is 10(5)-10(6) times greater for a student on that school bus than for a typical resident in the SoCAB. Vehicle self-pollution rate varies with bus window position (open or closed) and bus manufacture year. Our results can be used to develop cost-effective strategies to reduce children's exposure to school bus emissions. Our results indicate, for example, that even if emission reductions were many times more expensive per gram emitted for school buses than for an average vehicle, it would still be less expensive per gram inhaled by a student to reduce emissions from school buses than from an average vehicle. PMID- 15884350 TI - Decrease in net mercury methylation rates following iron amendment to anoxic wetland sediment slurries. AB - The rate of mercury methylation in anoxic wetland sediments is affected by the concentration of bioavailable complexes between Hg and sulfide. Previous research with pure bacterial cultures has shown that addition of ferrous iron reduces the net rate of mercury methylation by decreasing the concentration of dissolved sulfide. To assess the possibility of using this approach to decrease net mercury methylation in restored and constructed wetlands, laboratory experiments were conducted by adding Hg(II) and Fe(II) to sediments collected from six sites in five estuarine wetlands. Addition of 30 mM (0.07 mmol g(-1) or 3.9 mg g(-1)) Fe(II) decreased net mercury methylation relative to that of unamended controls by a factor of 2.1-6.6. In all cases, the observed decrease in net mercury methylation was accompanied by a decrease in the concentrations of sulfide and filterable mercury in the water overlying the sediments. When iron was added to one of the sediment samples at doses that were small relative to the concentration of sulfide present, net mercury methylation either increased slightly or was unaffected. Comparison of the results to speciation model predictions suggests that dissolved reduced sulfur-containing species play a role in the formation of uncharged, bioavailable Hg complexes. Although further research is needed to determine the long-term effect of iron amendment, these results suggest that iron addition decreases mercury methylation in authentic wetland sediments. PMID- 15884351 TI - Polycyclic aromatic hydrocarbon biodegradation rates: a structure-based study. AB - This study was designed to examine the role of molecular structure in determining the biodegradation rates of polycyclic aromatic hydrocarbons (PAHs). Laboratory experiments were performed in aqueous systems, and data were analyzed in a manner that allowed determination of first-order biodegradation rates independent of bioavailability limitations from physical-chemical processes. An aerobic mixed culture was used, which had been enriched on a broad range of PAHs. The 22 PAHs included in this study ranged in size from two to four rings and included compounds with 5-carbon rings and alkyl substituents. The range of observed biodegradation rates was only 1 order of magnitude, which is much less than that which is typically observed in the field. This supports the findings of other types of studies, which conclude that most of the observed variation in environmental PAH biodegradation rates comes from processes controlling the bioavailability of the compounds and not processes controlling uptake or biotransformation. Rate differences that were observed were attributable either to the presence of a 5-carbon ring or an alkyl substituent in an alpha position. Various molecular descriptors that might be expected to correlate with rate limiting steps in the biodegradation process were used in an attemptto develop a quantitative structure-activity relationship for the PAH biodegradation rates. No significant correlations were found, but rate limitation from interactions with the relevant enzymes remains a possibility. PMID- 15884352 TI - Oxidation of Fe(II) in rainwater. AB - Photochemically produced Fe(II) is oxidized within hours under environmentally realistic conditions in rainwater. The diurnal variation between photochemical production and reoxidation of Fe(II) observed in our laboratory accurately mimics the behavior of ferrous iron observed in field studies where the highest concentrations of dissolved Fe(ll) occur in afternoon rain during the period of maximum sunlight intensity followed by gradually decreasing concentrations eventually returning to early morning pre-light values. The experimental work presented here, along with the results of kinetics studies done by others, suggests thatthe primary process responsible for the decline in photochemically produced Fe(II) concentrations is oxidation by hydrogen peroxide. This reaction is first order with respect to both the concentrations of Fe(II) and H2O2. The second-order rate constant determined for six different authentic rain samples varied over an order of magnitude and was always less than or equal to the rate constant determined for this reaction in simple acidic solutions. Oxidation of photochemically produced ferrous iron by other oxidants including molecular oxygen, ozone, hydroxyl radical, hydroperoxyl/superoxide radical, and hexavalent chromium were found to be insignificant under the conditions present in rainwater. This study shows that Fe(II) occurs as at least two different chemical species in rain; photochemically produced Fe(II) that is oxidized over time periods of hours, and a background Fe(II) that is protected against oxidation, perhaps by organic complexation, and is stable against oxidation for days. Because the rate of oxidation of photochemically produced Fe(II) does not increase with increasing rainwater pH, the speciation of this more labile form of Fe(II) is also not controlled by simple hydrolysis reactions. PMID- 15884353 TI - Mechanism of interactions between Hg(II) and Demeton S: an NMR study. AB - The chemical fate of organophosphorus pesticides (OPs) has been proven to depend strongly on the chemistry of their aquatic environment. In particular, metal ions (and metal oxide surfaces) have been known to play an important role in the hydrolytic fate of OPs. Various postulates regarding the mechanism of metal-ion promoted hydrolysis of OPs have been made over the years. However, direct spectroscopic evidence to pinpoint the hydrolytic products and the exact interaction between metal ions and organophosphorus pesticides are still lacking. We report herein the first in-situ study of the interaction between an aqueous solution of Hg(II) and Demeton S using 1H- and 31P NMR spectroscopy. It was found that the interactions between Hg(II), a soft Lewis acid, and Demeton S tend to be a strong function of the aqueous speciation of Hg(II), and the bonding between Hg2+ and Demeton S does not involve the central P=O bond but rather Hg2+ bonds with the two sulfur atoms in the Demeton S side chain and subsequently stabilizes the Demeton S molecule, a phenomenon not previously reported for any metal ion-OP systems studied. On the basis of this study, generalizations regarding the nature of metal ion binding even within a given class of OPs (i.e., phosphorodithioates, phosphorothioates, phosphorothiolates, etc.) should be avoided or only made with extreme caution. PMID- 15884354 TI - Effects of pH and cationic and nonionic surfactants on the adsorption of pharmaceuticals to a natural aquifer material. AB - A wide range of pharmaceutical compounds have been identified in the environment, and their presence is a topic of growing concern, both for human and ecological health. Adsorption to aquifer materials and sediments is an important factor influencing the fate and transport of pharmaceutical compounds in the environment. Surfactants and other amphiphiles are known to influence the adsorption of many compounds and may be present in the environment from wastewaters or other sources. The work described here examines the adsorption of four pharmaceutical compounds, acetaminophen, carbamazepine, nalidixic acid, and norfloxacin, in the presence of a natural aquifer material. Adsorption was studied as a function of pH and in the presence and absence of two surfactants, cetylpyridinium chloride (CPC), a cationic surfactant, and Tergitol NP9, an ethoxylated nonionic surfactant. In the absence of surfactants, results indicate a 1-2 orders of magnitude variation in adsorption affinity with changing pH for each of the two quinolone pharmaceuticals (nalidixic acid and norfloxacin) but no measurable adsorption for carbamazepine or acetaminophen. In the presence of surfactants, adsorption of acetaminophen and carbamazepine was enhanced to extents consistent with compound hydrophobicity, while adsorption of nalidixic acid and norfloxacin was not. At high pH values, the anionic species of nalidixic acid exhibited enhanced adsorption in the presence of the cationic surfactant, CPC. PMID- 15884355 TI - Accounting for differences in estrogenic responses in rainbow trout (Oncorhynchus mykiss: Salmonidae) and roach (Rutilus rutilus: Cyprinidae) exposed to effluents from wastewater treatment works. AB - Effluents from wastewater treatment works (WwTWs) contain estrogenic substances that induce feminizing effects in fish, including vitellogenin (VTG) synthesis and gonadal intersex. Fish vary in their responsiveness to estrogenic effluents, but the physiological basis for these differences are not known. In this study, uptake of estrogen from two WwTW effluents (measured in hydrolyzed bile) and estrogenic response (VTG induction) were compared in a salmonid (rainbow trout, Onchorhynchus mykiss) and a cyprinid fish (roach, Rutilus rutilus). Immature rainbow trout were more responsive than maturing roach to the estrogenic effluents. The more potent of the two estrogenic effluents (containing between 24.3 and 104.1 ng estradiol-17beta equivalents/L [E2eq/L]) resulted in a 700-fold and 240-fold induction of plasma VTG in male and female trout, respectively, but only a 4-fold induction in roach (and in males only). The less potent effluent (varying between 4.1 and 6.8 ng E2eq/L) induced VTG in the trout only, with a 4 fold and 18-fold induction in males and females, respectively. In fish exposed to tap water, the estrogenicity of the hydrolyzed bile was 0.03+/-0.01 ng E2eq/microL (for both sexes in trout), 0.18+/-0.04 ng E2eq/microL in male roach, and 0.88+/-0.15 ng E2eq/microL in female roach. The higher bile content of estrogen in control roach reflected their more advanced sexual status (and thus higher endogenous estrogen) compared with the immature female trout. In trout maintained in effluents, the bile content of estrogen was 100-fold and 30-fold higher than controls at WwTW A and B, respectively. Bioconcentration factors (BCFs) for estrogenic activity in bile were between 16 344 and 46 134 in trout and between 3543 and 60 192 in roach (no gender differences were apparent). There were strong correlations between VTG induction and the estrogenic activity of bile extracts for both trout and roach. The results confirm that estrogenic contaminants bioconcentrate to a high degree in fish bile and that the level (and nature) of this accumulation may accountfor responsiveness to the endocrine disruptive effects of estrogenic effluents. Immature fish were the more appropriate life stage for quantifying estrogen exposure and uptake in bile, as they contain little circulating endogenous oestrogen compared with sexual maturing fish. The nature of the estrogenic contaminants is detailed in an accompanying paper. PMID- 15884356 TI - Interfacial interactions between Np(V) and manganese oxide minerals manganite and hausmannite. AB - Manganese oxides, present as minor phases in the vadose zone, have been previously shown to sequester large quantities of plutonium under environmental conditions. We are now continuing these studies with Np(V). Sorption onto manganite (MnOOH) and hausmannite (Mn3O4) at solid-to-solution ratios of 2.5-3.3 mg/mL has been studied as a function of neptunium concentration and pH. The sorption of Np increased as a function of pH for both minerals, attaining a maximum at neutral pH, and then decreased with increasing alkalinity. X-ray absorption fine structure spectroscopy (XAFS), taken at the Np L(III)-edge, has been used to determine the oxidation state of the sorbed Np. Our experimental results indicate reduction of the Np(V) because of interaction with the X-ray beam. These findings significantly impact the interpretation of results reported elsewhere on Np(V) investigated though the use of high-intensity X-ray beams. PMID- 15884357 TI - Enzymatic versus nonenzymatic conversions during the reduction of EDTA-chelated Fe(III) in BioDeNOx reactors. AB - Reduction of EDTA-chelated Fe(III) is one of the core processes in the BioDeNOx process, a chemically enhanced technique for biological NOx removal from industrial flue gases. The capacity of Escherichia coli, three mixed cultures from full scale methanogenic granular sludge reactors, one denitrifying sludge, and a BioDeNOx sludge to reduce Fe(III)EDTA- (25 mM) was determined at 37 and 55 degrees C using batch experiments. Addition of catalytic amounts of sulfide greatly accelerated Fe(III)EDTA- reduction, indicating that biological Fe(III)EDTA- reduction is not a direct, enzymatic conversion but an indirect reduction with involvement of an electron-mediating compound, presumably polysulfides. It is suggested that not thermophilic dissimilatory iron-reducing bacteria but reducers of elemental sulfur or polysulfides are primarily involved in the reduction of EDTA-chelated Fe(III) in BioDeNOx reactors. PMID- 15884358 TI - Cell-membrane damage and element leaching in transplanted Parmelia sulcata lichen related to ambient SO2, temperature, and precipitation. AB - Measurements were performed in lichen (Parmelia sulcata) transplants, to gain insight into the lichen vitality as possibly affected by both element deposition or lichen element content and further ambient atmospheric conditions (temperature, precipitation, SO2 levels). The electrical conductivity of rinsing solutions was used to assess cell-membrane damage in Parmelia sulcata in an experiment, which ran from August 2001 until August 2002. Element contents of the solutions and lichens were determined by inductively coupled plasma-optical emission spectrometry and by k0-instrumental neutron activation analysis, respectively. Factor analysis (MCATTFA) was used to determine grouping of elements of similar origin and/ or behavior. All in all, the data indicate that, apartfrom lichen Na and Cl levels, and for temperature and precipitation, no clear relationships with conductivity could be observed. Conductivity was mostly related to released Na, Cl, K, Mg, and Cs. On the basis of concentrations, Na, Cl, and K could be considered as largely determining the conductivity. The data suggest a different origin for K than that for Na and Cl: the latter two are most probably due to effects from sea salt sprays. Parmelia sulcata was sensitive enough to reflect appreciable ambient rises in air SO2 and resistant enough to recover afterward. MCATTFA on selected elements (K, Sc, Cu, V, As, and Sb) indicated the absence of any comparability between K and V, As and Sb, suggesting differences in origin and/or chemico-physical occurrence. Generally speaking, the present data suggest that the comparability of lichen vitality in large geographical areas may be limited and governed by the area's variability in temperature and precipitation rather than by variability in metal deposition rates. The leaching data on all elements and element groups, however, strongly suggest that wet deposition may also severely affect lichen elemental levels. This latter observation means that comparing outcomes for time or spatial series of lichen samples should be accompanied by a comparably careful monitoring of (preceding) ambient conditions. PMID- 15884359 TI - Reductive dechlorination of tetrachloroethene to trans-dichloroethene and cis dichloroethene by PCB-dechlorinating bacterium DF-1. AB - Polychlorinated biphenyls (PCBs) and chlorinated ethenes (CEs) are known to pollute sediment, soil, and groundwater. The anaerobic dechlorination of these compounds is an integral part of their biodegradation in polluted environments. We report for the first time the dechlorination of tetrachloroethene (PCE) and trichloroethene (TCE) by bacterium DF-1. This PCB and chlorobenzene dechlorinating bacterium dechlorinated PCE to TCE, which was then converted into trans-1,2-dichloroethene (trans-DCE) and cis-1,2-dichloroethene (cis-DCE). The ratio of trans-DCE to cis-DCE produced by the culture had a range of 1.2-1.7. Bacterium DF-1 has been enriched in co-culture with a desulfovibrio-like microorganism. PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of the 16S rRNA genes of the co-culture demonstrated that DF-1 was enriched during the dechlorination of PCE, PCB, and chlorobenzene. DF-1 was not detected in the absence of PCE dechlorination and the desulfovibrio-like organism, isolated in pure culture, did not dechlorinate PCE. This is the first identification of a microorganism capable of producing high amounts of trans-DCE from PCE and indicates that microorganisms such as DF-1 are a possible biological source of trans-DCE in the environment. PMID- 15884360 TI - Settling behavior of unpurified Cryptosporidium oocysts in laboratory settling columns. AB - The settling behavior of fresh and aged unpurified oocysts was examined in settling column suspensions with varied ionic strengths and concentrations of calcium and magnesium. Independent measurements of the size and density of unpurified oocysts were performed to determine a theoretical settling velocity for the test populations. Viability of the oocysts was assessed using a dye permeability assay. Latex microspheres were included to provide a standard by which to assess the settling conditions in the columns. Mean settling velocities for viable oocysts measured in this work were faster than predicted and faster than measured for purified oocysts in other work: 1.31 (+/-0.21) microm/s for viable oocysts from populations having a low percentage of viable oocysts and 1.05 (+/-0.20) microm/s for viable oocysts from populations with a high percentage of viable oocysts. Results were attributed to the higher than previously reported densities measured for oocysts in this study and the presence of fecal material, which allowed opportunity for particle agglomeration. Settling velocity of oocysts was significantly related to the viability of the population, particle concentration, ionic strength, and presence of calcium and magnesium in the suspending medium. Behavior of the latex microspheres was not entirely predictive of the behavior of the oocysts under the test conditions. Viable oocysts may have a greater probability of settling than previously assumed; however, nonviable, and especially nonintact, oocysts have the potential to be significantly transported in water. This work underscores the importance of assessing the viability of oocysts to predict their response to environmental and experimental conditions. PMID- 15884361 TI - Reduction of organically complexed ferric iron by superoxide in a simulated natural water. AB - Superoxide (and potentially its conjugate acid hydroperoxyl) is unique among the reactive oxygen species in that its standard redox potential in circumneutral natural waters potentially allows it to reduce ferric iron to the more soluble ferrous state. Here we have observed the superoxide/ hydroperoxyl-mediated reduction of ferric complexes with a variety of synthetic organic ligands and several complexes with natural organic matter (NOM), as well as freshly precipitated amorphous ferric oxyhydroxide, in bicarbonate buffered solutions at pH 8.1. From measurements of superoxide decay in the presence of the complexes, we calculated second-order rate constants for superoxide/ hydroperoxyl-mediated reduction that vary from (9.3+/-0.2) x 10(3) M(-1) s(-1) for the complex between Fe(III) and desferrioxamine B up to (1.9+/-0.2) x 10(5) M(-1) s(-1) for Fe(III) salicylate and (2.3+/-0.1) x 10(5) M(-1) s(-1) for one of the Fe(III)-NOM complexes. We also verified that ferrous iron was produced from superoxide/hydroperoxyl-mediated Fe(III) reduction using ferrozine to trap free Fe(II). Low yields of the ferrozine complex when compared to the measured rates of superoxide decay suggest that ferric complexes are reduced directlyto corresponding ferrous complexes, with much of the ferrous complex reoxidizing before it is able to release free ferrous iron. This is an important consideration for microorganisms, as the kinetics of trace metal uptake is typically governed by free ion activity. PMID- 15884362 TI - Cryogenic laser induced U(VI) fluorescence studies of a U(VI) substituted natural calcite: implications to U(VI) speciation in contaminated Hanford sediments. AB - Time-resolved laser-induced fluorescence spectroscopy (TRLFS) and imaging spectromicroscopy (TRLFISM) were used to examine the chemical speciation of uranyl in contaminated subsurface sediments from the U.S. Department of Energy (U.S. DOE) Hanford Site, Washington. Spectroscopic measurements for contaminant U(VI) were compared to those from a natural, uranyl-bearing calcite (NUC) that had been found via X-ray absorption spectroscopy (XAS) to include uranyl in the same coordination environment as calcium. Spectral deconvolution of TRLFS measurements on the NUC revealed the unexpected presence of two distinct chemical environments consistent with published spectra of U(VI)-substituted synthetic calcite and aragonite. Apparently, some U(VI) substitution sites in calcite distorted to exhibit a local, more energetically favorable aragonite structure. TRLFS measurements of the Hanford sediments NP4-1 and NP1-6 were similar to the NUC in terms of peak positions and intensity, despite a small CaCO3 content (1.0 to 3.2 mass %). Spectral deconvolution of the sediments revealed the presence of U(VI) in calcite and aragonite structural environments. A third, unidentified U(VI) species was also present in the NP1-6 sediment. TRLFISM measurements at multiple locations in the different sediments displayed only minor variation, indicating a uniform speciation pattern. Collectively, the measurements implied that waste U(VI), long-resident beneath the sampled disposal pond (32 y), had coprecipitated within carbonates. These findings have major implications for the solubility and fate of contaminant U(VI). PMID- 15884363 TI - Interaction of tetracycline with aluminum and iron hydrous oxides. AB - The effect of solution chemistry (pH, sorbate-to-sorbent ratio, ionic strength (/)) and reaction time on the sorption of tetracycline to the hydrous oxides of Al (HAO) and Fe (HFO) was examined. Sorption to HAO increased with increasing pH upto pH 7 (no such trend for HFO) above which it decreased at higher pH values for both the hydrous oxides. Experimental results indicate that ligand-promoted dissolution is occurring during tetracycline sorption to these hydrous oxides. Ligand-promoted dissolution was more significantfor HAO than HFO attributable to the difference in labile surface sites between these two sorbents. The ability of tetracycline to form strong complexes with Al and Fe will increase the solubility of these minerals. Sorption of tetracycline was quite rapid and equilibrium was achieved after 8 h. However, soluble metal (Me: Al or Fe) concentrations attained equilibrium only after 24 h. Ligand-promoted dissolution appears to be a two-step process; initially, 1:1 Me-tetracycline soluble complexes are formed and as the reaction progresses 2:1 complexes existed. Increasing / (from 0.01 to 0.5 M) decreased the sorption extent only at higher sorbate-to-sorbent ratios suggesting the dominance of inner-sphere type complexes at low equilibrium tetracycline concentrations. Spectroscopic evidence indicates that tricarbonylamide and carbonyl functional groups of tetracycline could be responsible for sorption to mineral surfaces. Our research findings will increase understanding of the environmental occurrence, fate, and transport characteristics of antibiotics, which are considered as emerging organic contaminants. PMID- 15884364 TI - Secondary organic aerosol formation by irradiation of 1,3,5-trimethylbenzene-NOx H2O in a new reaction chamber for atmospheric chemistry and physics. AB - A new environmental reaction smog chamber was built to simulate particle formation and growth similar to that expected in the atmosphere. The organic material is formed from nucleation of photooxidized organic compounds. The chamber is a 27 m3 fluorinated ethylene propylene (FEP) bag suspended in a temperature-controlled enclosure. Four xenon arc lamps (16 kW total) are used to irradiate primary gas components for experiments lasting up to 24 h. Experiments using irradiations of 1,3,5-trimethylbenzene-NOx-H2O at similar input concentrations without seed particles were used to determine particle number and volume concentration wall loss rates of 0.209+/-0.018 and 0.139+/-0.070 h(-1), respectively. The particle formation was compared with and without propene. PMID- 15884365 TI - Characterization of adsorption sites on aggregate soil samples using synchrotron X-ray computerized microtomography. AB - Synchrotron-source X-ray computerized microtomography (CMT) was used to evaluate the adsorptive properties of aggregate soil samples. A linear relationship between measured mean mass attenuation coefficient (sigma) and mass fraction iron was generated by imaging mineral standards with known iron contents. On the basis of reported stoichiometries of the clay minerals and identifications of iron oxyhydroxides (1), we calculated the mass fraction iron and iron oxyhydroxide in the intergranular material. The mass fractions of iron were estimated to range from 0.17 to 0.22 for measurements made at 18 keV and from 0.18 to 0.21 for measurements made at 26 keV. One aggregate sample also contained regions within the intergranular material with mass fraction iron ranging from 0.29 to 0.31 and from 0.33 to 0.36 for the 18 and 26 keV measurements, respectively. The mass fraction iron oxyhydroxide ranged from 0.18 to 0.35 for the low-iron intergranular material and from 0.40 to 0.59 for the high-iron intergranular material. Using absorption edge difference imaging with CMT, we visualized cesium on the intergranular material, presumably because of adsorption and possible exchange reactions. By characterizing the mass fraction iron, the mass fraction iron oxyhydroxide, and the adsorptive capacity of these soil mineral aggregates, we provide information useful for conceptualization, development, and parametrization of transport models. PMID- 15884366 TI - Some sources and sinks of monomethyl and inorganic mercury on Ellesmere Island in the Canadian High Arctic. AB - We identified some of the sources and sinks of monomethyl mercury (MMHg) and inorganic mercury (HgII) on Ellesmere Island in the Canadian High Arctic. Atmospheric Hg depletion events resulted in the deposition of Hg(II) into the upper layers of snowpacks, where concentrations of total Hg (all forms of Hg) reached over 20 ng/L. However, our data suggest that much of this deposited Hg(II) was rapidly photoreduced to Hg(0) which then evaded back to the atmosphere. As a result, we estimate that net wet and dry deposition of Hg(II) during winter was lower at our sites (0.4-5.9 mg/ha) than wet deposition in more southerly locations in Canada and the United States. We also found quite high concentrations of monomethyl Hg (MMHg) in snowpacks (up to 0.28 ng/L), and at times, most of the Hg in snowpacks was present as MMHg. On the Prince of Wales Icefield nearthe North Water Polynya, we observed a significant correlation between concentrations of Cl and MMHg in snow deposited in the spring, suggesting a marine source of MMHg. We hypothesize that dimethyl Hg fluxes from the ocean to the atmosphere through polynyas and open leads in ice, and is rapidly photolyzed to MMHgCl. We also found that concentrations of MMHg in initial snowmelt on John Evans Glacier (up to 0.24 ng/L) were higher than concentrations of MMHg in the snowpack (up to 0.11 ng/L), likely due to either sublimation of snow or preferential leaching of MMHg from snow during the initial melt phase. This springtime pulse of MMHg to the High Arctic, in conjunction with climate warming and the thinning and melting of sea ice, may be partially responsible for the increase in concentrations of Hg observed in certain Arctic marine mammals in recent decades. Concentrations of MMHg in warm and shallow freshwater ponds on Ellesmere Island were also quite high (up to 3.0 ng/L), leading us to conclude that there are very active regions of microbial Hg(II) methylation in freshwater systems during the short summer season in the High Arctic. PMID- 15884367 TI - Estimating partition coefficients for fuel-water systems: developing linear solvation energy relationships using linear solvent strength theory to handle mixtures. AB - In many environmental transport problems, organic solutes partition between immiscible phases that consist of liquid mixtures. To estimate the corresponding partition coefficients, we evaluated the efficacy of combining linear solvation energy relationships (LSERs) developed for pure 1:1 systems via application of linear solvent strength theory. In this way, existing LSERs could be extended to treat solute partitioning from gasoline, diesel fuel, and similar mixtures into contacting aqueous mixtures. Unlike other approaches, this method allowed prediction of liquid-liquid partition coefficients in a variety of fuel-water systems for a broad range of dilute solutes. When applied to 37 polar and nonpolar solutes partitioning between an aqueous mixture and 12 different fuel like mixtures (many including oxygenates), the root-mean-squared error was a factor of approximately 2.5 in the partition coefficient. This was considerably more accurate than application of Raoult's law for the same set of systems. Regulators and scientists could use this method to estimate fuel-water partition coefficients of novel additives in future fuel formulations and thereby provide key inputs for environmental transport assessments of these compounds. PMID- 15884368 TI - Five-stage environmental exposure assessment strategy for mixtures: gasoline as a case study. AB - A five-stage strategy is suggested for conducting an exposure assessment of mixtures that may contain numerous chemical components. The stages are: (1) determination of mixture composition and variability, (2) selection of component groups within the mixture and documentation of criteria used for this selection, (3) compilation of relevant property data for each group, (4) assessment of environmental fate of each group, and (5) assessment of environmental and human exposure to each group and to the mixture as a whole. A subsequent step is the assessment of environmental and/or human risk associated with the individual and aggregate exposure to each group. The approach is illustrated by application to gasoline, which is treated as 24 component groups or hydrocarbon blocks. Focusing on stages 2-4, the illustration shows that the groups display widely different environmental fates as a result of their different physicochemical properties, degradation half-lives, and mode-of-entry into the environment. As a result, the relative proportions of groups in each environmental medium (such as air and water) differ greatly from that of the original mixture. It is thus important to treat gasoline and similar mixtures as a number of component groups instead of as a single substance. A generic procedure is suggested in which the model is run for unit emissions of each component group to air, water, and soil. These results are compiled into matrices that can then be conveniently scaled to actual emission rates without rerunning the model. Methods for determining subsequent exposure and risk are also briefly outlined. PMID- 15884369 TI - Halogen substitution patterns among disinfection byproducts in the information collection rule database. AB - Disinfection byproduct halogen substitution patterns were examined using Information Collection Rule data. A new metric was developed to quantify and compare extents of specific halogen substitution in different byproduct classes. Bromine substitution in four byproduct classes ranged widely across 6565 samples but strong interdependencies existed between classes on an individual sample basis. Bromine substitution in dihaloacetic acids and trihalomethanes was comparable whereas bromine substitution in trihaloacetic acids and dihaloacetonitriles tended to be 10% lower and 60% higher than in trihalomethanes, respectively. However, these bromine substitution measurements were significantly impacted by censored data handling because reporting limit left-censoring increased with the number of bromine substituents in each class and minimum reporting levels varied between and within classes. Correlation among extents of bromine substitution in the four byproduct classes examined motivated their treatment as a multivariate response with application of a test for multivariate outliers. The test was effective at exposing inconsistencies indicative of data entry or analytical error. This work shows that disinfection byproduct data with full homologue series for multiple compound classes contain redundant information in the form of expected interdependencies among species. Application to quality assurance is demonstrated in this paper. Further applications to modeling and monitoring design are anticipated. PMID- 15884370 TI - Determination of the fractal dimension of microbial flocs from the change in their size distribution after breakage. AB - The fractal dimension is of ultimate importance in the description of the properties and behavior of microbial flocs in biological wastewater treatment systems. However, the methods for the measurement of the fractal dimension have not been well-established. In this study, a new method is developed to determine the fractal dimension of biological flocs in activated sludge (AS) suspensions. The new method utilizes the particle size distribution (PSD) of the sludge flocs measured by image processing under a microscope. Mechanical breakage is conducted for the sludge sample to create a change in the PSD. On the basis of the self similarity of fractal aggregates, the fractal dimension of the AS flocs can be calculated from a comparison between the original PSD and the altered PSD by breakage. It is determined that the sludge flocs grown in laboratory bioreactors with sludge ages of 5, 10, and 20 days have fractal dimensions of 2.07, 2.21, and 2.36. The sludge collected from a full-scale AS treatment plant has a fractal dimension of 1.99. This new method overcomes the deficiencies of other existing methods. It is easier to use and provides more reliable results in the determination of the fractal dimension of biological flocs and other similar aggregate samples. PMID- 15884371 TI - Degradation of biological weapons agents in the environment: implications for terrorism response. AB - We investigate the impact on effective terrorism response of the viability degradation of biological weapons agents in the environment. We briefly review the scientific understanding and modeling of agent environmental viability degradation. In general, agent susceptibility to viability loss is greatest for vegetative bacteria, intermediate for viruses, and least for bacterial spores. Survival is greatest in soil and progressively decreases in the following environments: textiles, water, hard surfaces, and air. There is little detailed understanding of loss mechanisms. We analyze the time behavior and sensitivity of four mathematical models that are used to represent environmental viability degradation (the exponential, probability, and first- and second-order catastrophic decay models). The models behave similarly at short times (<30 min for our example case) but diverge to significantly different values at intermediate to long times. Hence, for a release event in which the majority of atmospheric exposure or deposition occurs oververy short times, the current response models likely provide a good representation of the hazard. For longer time phenomena, including decontamination, the current model capabilities are likely insufficient. Finally, we implement each model in a simple numerical integration of anthrax dispersion, viability degradation, and dose response. Decay models spanning the current knowledge of airborne degradation result in vastly different predicted hazard areas. This confounds attempts to determine necessary medical and decontamination measures. Hence,the current level of understanding and representation of environmental viability degradation in response models is inadequate to inform appropriate emergency response measures. PMID- 15884372 TI - New analytical method for the determination of levoglucosan, polyhydroxy compounds, and 2-methylerythritol and its application to smoke and rainwater samples. AB - Biomass burning is an important source of smoke aerosol particles, which contain water-soluble inorganic and organic species, and thus have a great potential of affecting cloud formation, precipitation, and climate on global and regional scales. In this study, we have developed a new chromatographic method for the determination of levoglucosan (a specific tracer for biomass burning particles), related polyhydroxy compounds, and 2-methylerythritol (recently identified as isoprene oxidation product in fine aerosols in the Amazon) in smoke and in rainwater samples. The new method is based on water extraction and utilizes ion exclusion high-performance liquid chromatography (IEC-HPLC) separation and spectroscopic detection at 194 nm. The new method allows the analysis of wet samples, such as rainwater samples. In addition, aliquots of the same extracts can be used for further analyses, such as ion chromatography. The overall method uncertainty for sample analysis is 15%. The method was applied to the analysis of high-volume and size-segregated smoke samples and to rainwater samples, all collected during and following the deforestation fires season in Rondonia, Brazil. From the analysis of size-segregated samples, it is evident that levoglucosan is a primary vegetation combustion product, emitted mostly in the 0.175-1 microm size bins. Levoglucosan concentrations decrease below the detection limit atthe end of the deforestation fires period, implying that it is not present in significant amounts in background Amazon forest aerosols. The ratio of daytime levoglucosan concentration to particulate matter (PM) concentration was about half the nighttime ratio. This observation is rationalized by the prevalence of flaming combustion during day as opposed to smoldering combustion during night. This work broadens the speciation possibilities PMID- 15884373 TI - Partition of endocrine-disrupting chemicals between colloids and dissolved phase as determined by cross-flow ultrafiltration. AB - Cross-flow ultrafiltration (CFUF) was developed first for the isolation of natural colloids and subsequently for determining the partition of selected endocrine-disrupting chemicals (EDCs) between river colloids and dissolved phase. In this study, a 1-kDa Millipore Pellicon 2 cartridge type CFUF system was validated using a range of molecular probes spiked in natural waters. Results show that good retention (>80%) of high molecular weight (HMW, >1 kDa) molecules and low retention of low molecular weight (LMW, <1 kDa) molecules can be achieved at high concentration factor (cf) values in sampling mode or over long time scales in recirculation mode. The interactions between aquatic colloids and EDCs were studied by mixing EDCs, water, and colloids previously isolated by CFUF for a certain duration, followed by the separation of the target compounds between the truly dissolved and colloid-bound phases by CFUF and analysis by gas chromatography-mass spectrometry (GC-MS). The kinetics of EDCs binding to colloids were relatively rapid, reaching equilibrium within 5 min. The mass balance of chosen EDCs through CFUF system was fully investigated, with good recovery for the relatively polar EDCs such as estrone and 17beta-estradiol. On the basis of EDC sorption by colloids, the partition coefficient normalized to colloidal organic carbon content (Kcoc) was 8.85 x 10(3), 1.50 x 10(4), 8.85 x 10(3), 4.87 x 10(4), and 1.59 x 10(4) mL/g for bisphenol A, estrone, 17beta estradiol, 17alpha-ethynylestradiol, and 16alpha-hydroxyestrone, respectively, which are comparable with the values reported in the literature. In addition, it has been shown that the Kcoc values of EDCs were relatively independent of their octanol-water partition coefficient (Kow) values, suggesting the important role of different binding mechanisms other than nonspecific hydrophobic interaction between EDCs and natural colloids. As the CFUF-GC-MS method can be used to quantify very low concentration of pollutants and is not limited to fluorescent compounds, it has the potential to be a widely applicable separation/analytical tool for determining the partition of organic pollutants between colloidal materials and dissolved phase. PMID- 15884374 TI - In vivo visual reporter system for detection of estrogen-like substances by transgenic medaka. AB - Detection of endocrine disrupting chemicals, in particular, environmental estrogens with living organisms, has many advantages if compared to chemical analysis. The screening of novel pollutants with meaningful endpoints, the integration of uptake, bioconcentration, and excretion as well as the evaluation of endocrine disrupting effects with respect to toxicity require in vivo biotests for estrogen-like substances (ELSs). Critical disadvantages of whole organism biotests are their low sensitivity and the need for laborious and time-consuming work. To overcome these problems, we have developed a transgenic medaka strain harboring the green fluorescence protein (GFP) gene driven by choriogenin H gene regulatory elements. Choriogenin H is an egg envelope protein induced by estrogens in the liver. With yolk sac larvae of this strain, GFP induction in liver was observed 24 h after onset of aqueous exposure to 0.63 nM 17beta estradiol (E2), 0.34 nM ethynylestradiol, or 14.8 nM estrone. Furthermore, concentrated sewage treatment effluent induced GFP expression. Comparison of E2 equivalents estimated by GFP-induction in transgenic medaka, a YES assay, and GC/MS showed detection limits in the same order of magnitude. These results indicated that the sensitivity of the transgenic medaka strain was sufficient for application as an alternative model in monitoring environmental water samples for ELSs. PMID- 15884375 TI - Microbenthic chamber with microelectrode for in-situ determination of fluxes of dissolved S(-II), I-, O2, Mn, and Fe. AB - A 2-mL microbenthic chamber was fitted with a microelectrode for the in-situ determination of benthic fluxes of S(-II), I-, O2, Mn(II), and Fe(I). Detection was by voltammetry using a battery operated potentiostat and a gold microelectrode. The chamber was fitted on a Perspex plate to be placed on sediments. Because of the small chamber volume, benthic fluxes could be determined in a few hours rather than days, without the need for sample extraction. Tests on homogenized sediments in the laboratory showed fluxes of 17.1+/-1.8 nmoles cm(-2) min(-1) S(-II) and 1.3+/-0.2 nmoles cm(-2) min(-1) Mn. Benthic fluxes of oxygen and iodide were determined in situ in the field. The oxygen flux was negative (consumption) at a rate of -4.9+/-0.5 nmoles cm(-2) min( 1) O2. The I- flux was initially negative in oxygenated waters at a rate of -30+/ 3 pmoles cm(-2) min(-1) and subsequently turned positive to a rate of 12+/-1 pmoles cm(-2) min(-1) when the oxygen concentration dropped. The rate of change in the microbenthic chamber was sufficiently quick to complete a flux measurement within minutes. PMID- 15884376 TI - Quantitative RT-PCR methods for evaluating toxicant-induced effects on steroidogenesis using the H295R cell line. AB - Gene expression profiles show considerable promise for the evaluation of the toxic potential of environmental contaminants. For example, any alterations in the pathways of steroid synthesis or breakdown have the potential to Cause endocrine disruption. Therefore monitoring these pathways can provide information relative to a chemical's ability to impact endocrine function. One approach to monitoring these pathways has been to use a human adrenocortical carcinoma cell line (H295R) that expresses all the key enzymes necessary for steroidogenesis. In this study we have further developed these methods using accurate and specific quantification methods utilizing molecular beacon-based quantitative RT-PCR (Q-RT PCR). The assay system was used to analyze the expression patterns of 11 steroidogenic genes in H295R cells. The expression of gene transcripts was measured using a real-time PCR system and quantified based on both a standard curve method using a dilution series of RNA standards and a comparative Ct method. To validate the optimized method, cells were exposed to specific and nonspecific model compounds (inducers and inhibitors of various steroidogenic enzymes) for gene expression profiling. Similar gene expression profiles were exhibited by cells treated with chemicals acting through common mechanisms of action. Overall, our findings demonstrated that the present assay can facilitate the development of compound-specific response profiles, and will provide a sensitive and integrative screen for the effects of chemicals on steroidogenesis. PMID- 15884377 TI - Evaluation of a novel malathion immunoassay for groundwater and surface water analysis. AB - Malathion is an organophosphorus insecticide commonly used in crops and indoor applications. Negative effects of malathion on human health and ecosystems are of growing concern. In this work, novel malathion haptens are synthesized to develop an ELISA screening method. The immunoassay is based on a conjugate-coated format and shows a limit of detection of 0.11 ng/mL, an IC50 of 1.58 ng/mL, a dynamic range between 0.23 and 10.94 ng/ mL, and a cross-reactivity of <2% with structurally related compounds. The developed ELISA has been used to quantify malathion in groundwater and surface water samples. The good recoveries achieved and the agreement with those given by the GC-MS reference method indicate the potential of the assay for environmental monitoring of malathion in natural waters without purification or preconcentration steps. The effect of dissolved organic matter (humic acids) on the ELISA was evaluated, resulting in the conclusion that the immunoassay can be successfully performed in surface water samples with a humic acid content up to 10 mg/L, without sample pretreatment. Samples with a high humic acid content can be analyzed through ELISA after solid phase extraction, eluting with acetone or acetonitrile. PMID- 15884378 TI - Solid-phase microextraction measurement of parent and alkyl polycyclic aromatic hydrocarbons in milliliter sediment pore water samples and determination of K(DOC) values. AB - The U.S. Environmental Protection Agency (EPA) narcosis model for benthic organisms in polycyclic aromatic hydrocarbon (PAH) contaminated sediments requires the measurement of 18 parent PAHs and 16 groups of alkyl PAHs ("34" PAHs) in pore water with desired detection limits as low as nanograms per liter. Solid-phase microextraction (SPME) with gas chromatographic/mass spectrometric (GC/ MS) analysis can achieve such detection limits in small water samples, which greatly reduces the quantity of sediment pore water that has to be collected, shipped, stored, and prepared for analysis. Four sediments that ranged from urban background levels (50 mg/kg total "34" PAHs) to highly contaminated (10 000 mg/kg total PAHs) were used to develop SPME methodology for the "34" PAH determinations with only 1.5 mL of pore water per analysis. Pore water was obtained by centrifuging the wet sediment, and alum flocculation was used to remove colloids. Quantitative calibration was simplified by adding 15 two- to six-ring perdeuterated PAHs as internal standards to the water calibration standards and the pore water samples. Response factors for SPME followed by GC/MS were measured for 22 alkyl PAHs compared to their parent PAHs and used to calibrate for the 18 groups of alkyl PAHs. Dissolved organic carbon (DOC) ranging from 4 to 27 mg/L had no measurable effect on the freely dissolved concentrations of two- and three ring PAHs. In contrast, 5-80% of the total dissolved four- to six-ring PAHs were associated with the DOC rather than being freely dissolved, corresponding to DOC/water partitioning coefficients (K(DOC)) with log K(DOC) values ranging from 4.1 (for fluoranthene) to 5.6 (for benzo[ghi]perylene). However, DOC-associated versus freely dissolved PAHs had no significant effect on the total "34" PAH concentrations or the sum of the "toxic units" (calculated bythe EPA protocol), since virtually all (86-99%) of the dissolved PAH concentrations and toxic units were contributed by two- and three-ring PAHs. PMID- 15884379 TI - Solid waste removes toxic liquid waste: adsorption of chromium(VI) by iron complexed protein waste. AB - The leather processing industry generates huge amounts of wastes, both in solid and liquid form. Fleshing from animal hides/skins is one such waste that is high in protein content. In this study, raw fleshing has been complexed with iron and is used for removal of chromium(VI). The effect of pH and the initial concentration of chromium(VI) on the removal of Cr(IV) by iron treated fleshing is presented. Iron treatment is shown to greatly improve adsorption of the fleshing for hexavalent chromium. The ultimate adsorption capacity of iron treated fleshing is 51 mg of chromium(VI) per gram of fleshing. That of untreated fleshing is 9 mg/g such that iron treatment increases the adsorption capacity of fleshing by 10-fold. The measured adsorption kinetics is well described by a pseudo-second-order kinetic model. The uptake of chromium(VI) by fleshing is best described by the Langmuir adsorption isotherm model. X-ray photoelectron spectroscopic (XPS) studies show that the iron is incorporated into the protein matrix. Shifts in XPS spectra suggest that dichromate binding occurs with iron at active adsorption sites and that iron treated fleshing removes chromium(VI) without reducing it to chromium(III). PMID- 15884380 TI - Formation of soluble organo-chromium(III) complexes after chromate reduction in the presence of cellular organics. AB - Microbial reduction of hexavalent chromium [Cr(VI)] to trivalent chromium [Cr(III)] has been investigated as a method for bioremediation of Cr(VI) contaminated environments. The produced Cr(III) is thought to be insoluble Cr(OH)3; however, recent reports suggested a more complex fate of Cr(III). A bacterial enzyme system, using NADH as the reductant, converts Cr(VI)to a soluble NAD+-Cr(III) complex, and cytochrome c-mediated Cr(VI) reduction produces cytochrome c-Cr(III) adducts. In this study, Cr(VI) reduction in the presence of cellular organic metabolites formed both soluble and insoluble organo-Cr(III) end products. Several soluble end-products were characterized by absorbance spectroscopy and electron paramagnetic resonance spectrometry as organo-Cr(III) complexes, similar to the known ascorbate-Cr(III) complex. The complexes remained soluble and stable upon dialysis against distilled H20 and over a broad pH range. The ready formation of stable organo-Cr(III) complexes suggests that organo Cr(III) complexes are rather common, likely representing an integral part of the natural cycling of chromium. Thus, organo-Cr(III) complexes may account for the mobile form of Cr(II) detected in the environment. PMID- 15884381 TI - Formation and characterization of aerobic granules in a sequencing batch reactor treating soybean-processing wastewater. AB - Aerobic granules were cultivated in a sequencing batch reactor (SBR) fed with soybean-processing wastewater at 25+/-1 degrees C and pH 7.0+/-0.1. The granulation process was described via measuring the increase of sludge size. The formation of granules was found to be a four-phase process, that is, acclimating, shaping, developing, and maturated. A modified Logistic model could well fit with the granule growth by diameter and could be employed to estimate the maximum diameter, lag time, and specific diameter growth rate effectively. Both normal and log-normal distributions proved to be applicable to model the diameter distribution of the granules. The granule-containing liquor was shear thinning, and their rheological characteristics could be described by using the Herschel Buckley equation. The suspended solids concentration, pH, temperature, diameter, settling velocity, specific gravity, and sludge volume index all had an effect on the apparent viscosity of the mixed liquor of granules. The matured granules had fractal nature with a fractal dimension of 1.87+/-0.34. Moreover, 83% of matured granules were permeable with fluid collection efficiencies over 0.034. As compared to activated sludge flocs, the aerobic granules grown on the soybean processing wastewater had better settling ability, mass transfer efficiency, and bioactivity. PMID- 15884382 TI - Optimization of coagulation-flocculation process for palm oil mill effluent using response surface methodology. AB - The coagulation-flocculation process incorporated with membrane separation technology will become a new approach for palm oil mill effluent (POME) treatment as well as water reclamation and reuse. In our current research, a membrane pilot plant has been used for POME treatment where the coagulation-flocculation process plays an important role as a pretreatment process for the mitigation of membrane fouling problems. The pretreated POME with low turbidity values and high water recovery are the main objectives to be achieved through the coagulation flocculation process. Therefore, treatment optimization to serve these purposes was performed using jar tests and applying a response surface methodology (RSM) to the results. A 2(3) full-factorial central composite design (CCD) was chosen to explain the effect and interaction of three factors: coagulant dosage, flocculent dosage, and pH. The CCD is successfully demonstrated to efficiently determine the optimized parameters, where 78% of water recovery with a 20 NTU turbidity value can be obtained at the optimum value of coagulant dosage, flocculent dosage, and pH at 15 000 mg/L, 300 mg/L, and 6, respectively. PMID- 15884383 TI - Bench-scale investigation of permanganate natural oxidant demand kinetics. AB - A vital design parameter for any in situ chemical oxidation system using permanganate (MnO4-) is the natural oxidant demand (NOD), a concept that represents the consumption of MnO4- by the naturally present reduced species in the aquifer solids. The data suggest that the NOD of the aquifer material from Canadian Forces Base Borden used in our study is controlled by a fast or instantaneous reaction captured by the column experiments, and a slower reaction as demonstrated by both column and batch test data. These two reaction rates may be the result of the reaction of MnO4- with at least two different reduced species exhibiting widely different rates of permanganate consumption (fast rate >7 g of MnO4- as KMnO4/kg/day and slow rate of approximately 0.005 g/kg/day), or a physically/chemically rate-limited single species. The slow NOD reaction prevented fulfillment of the ultimate NOD during the days- to months-long batch experiments and allowed significant early MnO4- breakthrough (>98%) during transport in the column experiments. A large fraction of the organic carbon resisted oxidation over the 21-week duration of the batch experiments. This result demonstrates that NOD estimated from total organic carbon measurements can significantly overpredict the NOD value required in the design of an in situ chemical oxidation application. PMID- 15884384 TI - Selective catalytic reduction of nitrogen oxides from exhaust of lean burn engine over in-situ synthesized Cu-ZSM-5/cordierite. AB - ZSM-5 zeolite was in-situ synthesized successfully on the surface of honeycomb cordierite substrate, certified by XRD and SEM techniques. Good thermal and hydrothermal stability of ZSM-5/cordierite could be obtained because of the in situ synthesis method. Copper ion-exchanged ZSM-5/cordierite was studied as catalyst for selective catalytic reduction of nitrogen oxides. For practical reasons, the catalytic testing experiments were preformed on a real lean burn engine. Unburned hydrocarbons and carbon monoxide in the exhaust were directly used as reductants for NOx reduction. Cu-ZSM-5/cordierite exhibited high catalytic activity, and at 673 K the maximal NOx conversion to N2 could reach 50% at gas hourly space velocity (GHSV) of about 25 000 h(-1). Hydrocarbons and carbon monoxide in the exhaust could also be purified at the same time. As expected, Cu-ZSM-5/cordierite catalyst exhibited good duration and antipoison properties. When traces of lanthanum were introduced to Cu-ZSM-5/cordierite catalyst as a modifier, the activated temperature of the catalyst could be decreased and the temperature window with high NOx conversions broadened. Cu-ZSM 5/ cordierite and LaCu-ZSM-5/cordierite were thought as promising automobile exhaust catalysts under lean conditions. Copper is the main active component in the Cu-ZSM-5/ cordierite catalyst and Cu(II), which was found in the catalyst during the proceeding of reaction by XPS, is thought to be essential. On the basis of this, deNOx process over Cu-ZSM-5/cordierite is also discussed. PMID- 15884385 TI - Laboratory studies of electrochemical treatment of industrial azo dye effluent. AB - Removal of color and reduction of chemical oxygen demand (COD) in an industrial azo dye effluent containing chiefly reactive dyes were investigated under single pass conditions at a dimensionally stable anode (DSA) in a thin electrochemical flow reactor at different current densities, flow rates, and dilutions. With 50% diluted effluent, decolorization was achieved up to 85-99% at 10-40 mA/ cm2 at 5 mL/min flow rate and 50-88% at 30-40 mA/ cm2 at high (10-15 mL/min) flow rates. The COD reduction was maximum (81%) at 39.9 mA/cm2 or above when solution electrode contact time (Ct) was as high as 21.7 s/cm2 and decreased as Ct declined at a given current density. Cyclic voltammetric studies suggesting an indirect oxidation of dye molecules over the anode surface were carried out at a glassy carbon electrode. The effect of pH on decolorization and COD reduction was determined. An electrochemical mechanism mediated by OCl- operating in the decolorization and COD reduction processes was suggested. The effluent was further treated with NaOCI. The oxidized products from the treated effluents were isolated and confirmed to be free from chlorine-substituted products by IR spectroscopy. From the apparent pseudo-first-order rate data, the second-order rate coefficients were evaluated to be 2.9 M(-1) s(-1) at 5 mL/ min, 76.2 M(-1) s(-1) at 10 mL/min, and 156.1 M(-1) s(-1) at 15 mL/ min for color removal, and 1.19 M(-1) s(-1) at 5 mL/min, 1.79 M(-1) s(-1) at 10 mL/min, and 3.57 M(-1) s(-1) at 15 mL/min for COD reduction. Field studies were also carried out with a pilot scale cell at the source of effluent generation of different plants corresponding to the industry. Decolorization was achieved to about 94-99% with azo dye effluents at 0.7-1.0 L/min flow costing around Indian Rupees 0.02-0.04 per liter, and to about 54-75% in other related effluents at 0.3-1.0 L/min flow under single pass conditions. PMID- 15884386 TI - Sorption of 243Am(III) to multiwall carbon nanotubes. AB - Carbon nanotubes have attracted great interest in multidisciplinary study since their discovery. Herein, radionuclide 243Am(III) sorption to uncapped multiwall carbon nanotubes (MWCNTs) was carried out at 20+/-2 degrees C in 0.01 and 0.1 M NaClO4 solutions. Effects of 243Am(III) solution concentration, ionic strength, and pH on 243Am(III) sorption to MWCNTs were also investigated. The sorption is strongly dependent on pH values and weakly dependent on the ionic strength in the experimental conditions. The results show that MWCNTs can adsorb 243Am(III) with extraordinarily high efficiency by forming very stable complexes. Chemisorption or chemicomplexation is the main mechanism of 243Am(III) sorption on the surface of MWCNTs. MWCNTs can be a promising candidate for the preconcentration and solidification of 243Am(III) or its analogue lanthanides and actinides from large volumes of aqueous solution, as required for remediation purposes, and perhaps also as a sorbent for the removal of heavy metal ions from the industry wastewater. PMID- 15884387 TI - Fluidized bed combustion systems integrating CO2 capture with CaO. AB - Capturing CO2 from large-scale power generation combustion systems such as fluidized bed combustors (FBCs) may become important in a CO2-constrained world. Using previous experience in capturing pollutants such as SO2 in these systems, we discuss a range of options that incorporate capture of CO2 with CaO in FBC systems. Natural limestones emerge from this study as suitable high-temperature sorbents for these systems because of their low price and availability. This is despite their limited performance as regenerable sorbents. We have found a range of process options that allow the sorbent utilization to maintain a given level of CO2 separation efficiency, appropriate operating conditions, and sufficiently high power generation efficiencies. A set of reference case examples has been chosen to discuss the critical scientific and technical issues of sorbent performance and reactor design for these novel CO2 capture concepts. PMID- 15884388 TI - Longevity of granular iron in groundwater treatment processes: corrosion product development. AB - Permeable reactive barriers employing iron as a reactive surface have received extensive attention. A remaining issue, however, relates to their longevity. As an integral part of a long-term column study conducted to examine the influence of inorganic cosolutes on iron reactivity toward chlorinated solvents and nitroaromatic compounds, Master Builder iron grains were characterized via scanning and transmission electron microscopy, electron energy loss spectroscopy (EELS), micro-Raman spectroscopy, and X-ray diffraction. Prior to exposure to carbonate solutions, the iron grains were covered by a surface scale that consisted of fayalite (Fe2SiO4), wustite (FeO), magnetite (Fe3O4), maghemite (gamma-Fe2O3), and graphite. After 1100 days of exposure to solutions containing carbonate, other inorganic solutes, and organic contaminants, the wustite, fayalite, and graphite of the original scale partially dissolved, and magnetite and iron carbonate hydroxide (Fe3(OH)2.2CO3) precipitated on top of the scale. Raman results indicate the presence of green rust (e.g., [Fe4(2+)Fe2(3+)(OH)12] [CO3 x 2H2O]) toward the column outlet after 308 days of operation, although this mineral phase disappears at longer operation times. Grains extracted from a column exposed to a high concentration (20 mM) of sodium bicarbonate were more extensively weathered than those from columns exposed to 2 mM sodium bicarbonate. An iron carbonate hydroxide layer up to 100 microm thick was observed. Even though EELS analysis of iron carbonate hydroxide indicates that this is a redox active phase, the thickness of this layer is presumed responsible for the previously observed decline in the reactivity of this column relative to low bicarbonate columns. A silica-containing feed resulted in reduced reactivity toward TCE. Grains from this column had a strong enrichment of silicon in the precipitates, although no distinct silica-containing mineral phases were identified. The substitution of 2 mM calcium carbonate for 2 mM sodium bicarbonate in the feed did not produce a measurable reactivity loss, asthe discrete calcium carbonate precipitates that formed in this system did not severely restrict access to the reactive surface. PMID- 15884389 TI - Addition of activated carbon to sediments to reduce PCB bioaccumulation by a polychaete (Neanthes arenaceodentata) and an amphipod (Leptocheirus plumulosus). AB - This work examines the effects of adding coke or activated carbon on the bioavailability of polychlorinated biphenyls (PCBs) in contaminated sedimentfrom South Basin at Hunters Point, San Francisco Bay. We show with 28-day sediment exposure tests that PCB bioaccumulation in a polychaete (Neanthes arenaceodentata) is reduced by 82% following 1-month contact of sediment with activated carbon and by 87% following 6-months contact of sediment with activated carbon. PCB bioaccumulation in an amphipod (Leptocheirus plumulosus) is reduced by 70% following 1-month contact of sediment with activated carbon and by 75% after 6-months contact of sediment with activated carbon. Adding coke had a negligible effect on reducing PCB bioaccumulation, probably because of the low specific surface area and the slow kinetics of PCB diffusion intothe solid coke particles. Reductions in congener bioaccumulation with activated carbon were inversely related to congener Kow, suggesting that the efficacy of activated carbon is controlled by the mass-transfer rate of PCBs from sediment and into activated carbon. We find that reductions in aqueous PCB concentrations in equilibrium with the sediment were similar to reductions in PCB bioaccumulation. While no lethality was observed following activated carbon addition, growth rates were reduced by activated carbon for the polychaete, but not for the amphipod, suggesting the need for further study of the potential impacts of activated carbon on exposed communities. The study suggests that treatment of the biologically active layer of contaminated sediments with activated carbon may be a promising in-situ technique for reducing the bioavailability of sediment associated PCBs and other hydrophobic organic compounds. PMID- 15884390 TI - Laboratory study of treatment of trichloroethene by chemical oxidation followed by bioremediation. AB - Studies were conducted with columns containing soil and emplaced trichloroethene (TCE) to investigate the potential for TCE source zone remediation with chemical oxidation followed by biologically mediated reductive dehalogenation. Following permanganate flushing of four columns, which resulted in rapid but incomplete removal of TCE DNAPL, no biological activity was observed following the addition of distilled water amended with ethanol and acetate, including two of the four columns that were bioaugmented with a TCE-dechlorinating microbial culture. Flushing with unsterilized site groundwater led to consumption of acetate and ethanol, accompanied by manganese reduction and methanogenesis. Reductive dechlorination of TCE to cis-1,2-dichloroethene (cis-DCE) followed the onset of ethanol and acetate biodegradation in bioaugmented columns only. Partial dechlorination of TCEto ethene was observed only in one of the bioaugmented columns after it was inoculated for a third time. At the end of the study (290 days), a trace amount of cis-DCE was observed in one of the two columns which was not bioaugmented. Reduced conditions created by biostimulation were also conducive to reduction of Mn(IV) from MnO2 in both bioaugmented and nonbioaugmented columns resulting in an increased dissolved manganese (Mn2+) concentration in groundwater. PMID- 15884391 TI - Assessment of the energy dissipation parameters inside the draft tube of a liquid spout-fluid bed. AB - Spouted beds are fluid-particle contactors in which the fluid is introduced centrally through a nozzle instead of a distributor plate, resulting in a regular particle circulation pattern. To assess the suitability of such sytems to environmental engineering applications such as filter backwashing and biofilm systems, a priori knowledge of the energy dissipation parameters is essential. A new model is developed for evaluating the energy dissipation parameters inside the draft tube of spout-fluid beds. The shear stress, velocity gradient, and turbulence fluctuation parameters in the draft tube of a liquid spout-fluid bed are calculated with the help of an energy equation for flows carrying suspensions and the experimentally determined pressure losses inside the draft tube and compared with results for particulately fluidized beds. A spout-fluid bed with a draft tube provides higher shear stress inside the draft tube than a fluidized bed. The mean velocity gradient in the draft tube is comparable to and higher than in a fluidized bed and increases with solids fraction. The turbulence dissipation coefficient decreases very slightlywith increasing solids fraction for both systems. Consequently, according to the model calculations, a spout fluid bed with a draft tube can be an alternative to the classical fluidized bed filter backwashing system. PMID- 15884392 TI - Electrodialytic removal of Cu, Zn, Pb, and Cd from harbor sediment: influence of changing experimental conditions. AB - Electrodialytic remediation (EDR) was used to remove Cu, Zn, Pb, and Cd from contaminated harbor sediment. Extraction experiments were made prior to EDR, and the metal desorption was pH dependent but not liquid-to-solid ratio (L/S) dependent. The desorption order was Cd > Zn > Pb > Cu. Electrodialytic experiments were made with HCl as desorbing agent in a sediment suspension, which was stirred during EDR. Effects of different current strengths and L/S ratios on the heavy metal removal were investigated on wet and air-dried sediment. The effects of drying the sediment were negligible for the removal of Cu, Zn, and Pb, probably due to oxidation of the sediments during stirring. Contrary, Cd removal was lower in the wet sediment as compared to the air-dried. The heavy metal removal was influenced by higher current strengths and varying L/S ratios. The highest removal obtained was in an experiment with dry sediment (L/S 8) and a 70 mA applied current that lasted 14 days. These experimental conditions were thereafter used to remediate more strongly contaminated sediments. Regardless of the initial heavy metal concentrations in the sediments, 67-87% Cu, 79-98% Cd, 90 97% Zn, and 91-96% Pb were removed. PMID- 15884393 TI - [Dizziness-vertigo. Alarming symptoms in vertebrobasilar ischemia. Part II]. AB - Dizziness and vertigo--like headache--are the most common complaints which lead patients to visit the doctor. In spite of the headache--which may be primary (e.g. migraine) or symptomatic--dizziness and vertigo do not appear to be a separate nosologic entity but rather the symptoms of several neurological disorders. For differential diagnosis, interdisciplinary thinking and action is needed because the vestibular, neurological and psychiatric disorders might have a common role in the development of symptoms and further overlapping can also occur. The vascular disorders of the vertebrobasilar system are discussed in detail in this review. The importance, occurrence and causes of vertigo as a warning symptom is in the focus. The author draws attention to life-threatening conditions with acute onset in cases of the posterior scale ischemia and emphasizes the importance of the correct and early diagnosis. The author tries to clear up the nihilistic aspect in treating of stroke and stresses the necessity of thrombolysis and interventional radiological procedures which may be the only chance for the recovery of the patients. The pharmacological prevention of recurrent vascular events is also important and obligatory for the clinicians. PMID- 15884394 TI - [Chronic inflammatory polyneuropathies]. AB - There is an increasing number of peripheral nerve disorders with inflammatory and immune mechanisms involved. The precise diagnosis is of utmost importance, since these patients can be successfully treated. Unfortunately, there is no specific marker for any disease of this group. The diagnosis therefore relies on the appropriate consideration of the clinical, neurophysiological and laboratory data, which requires in-depth knowledge of these diseases. In this paper we review the diagnostic criteria and treatment strategies for the major types of chronic inflammatory polyneuropathies. PMID- 15884395 TI - The perisylvian epileptic network. A unifying concept. AB - In this work the authors provide evidences for a unifying concept of the syndromes of benign focal childhood epilepsies, Landau-Kleffner syndrome, and electrical status epilepticus in sleep treating them as a spectrum of disorders with a common transient, age dependent, non lesional, genetically based epileptogenic abnormality, the nature of which is still not known. The electro clinical features of these syndromes are congruent with the different degree involvement of the perisylvian cognitive network and with the involvement of the thalamo-cortical associative system of variable degree. These epilepsies are characterized by the abundance of regional epileptiform discharges in sharp contrast with the rare and in several cases lacking seizures. The nature and severity of interictal cognitive symptoms are closely related to localization within the network and amount of epileptic interictal discharges. Spike-wave discharges are attributed to an alternation of overexcitation (spikes) and overinhibition (waves). The recurrent overinhibition represented by the wave of the discharges may interfere with the continuous depolarization of the cells of a large population of neurons, which is a requirement of the overt seizures. The overinhibition also interfere with cognitive processes which are correlated with the continuous presence of the fast (gamma) activity, binding the required cortical areas. Hence the recurrent inhibition works against the existence of the binding fast frequency activity. This is the assumed reason for the co-existence of the relative lack of overt seizures and in the same time for the frequently observed epileptogenic cognitive deficit symptoms ("cognitive epilepsies"). The time course of these syndromes overlaps with important developmental milestones. The frequent epileptic discharges alters the evolution of the perisylvian network developing late after early childhood and is very vulnerable for any interference in this imprinting time for speech and other cognitive functions. This spectrum of disorders represents a type of age linked, mild to severe 'epileptic encephalopathy' limited to the perisylvian network, where the cognitive impairment is caused by epileptic discharges interfering with cognitive development. PMID- 15884396 TI - [Activity of natural coagulation inhibitor proteins in the acute phase of ischaemic stroke]. AB - INTRODUCTION: Decreased activity of natural anticoagulants (antithrombin-lll, protein C, protein S) rarely causes cerebral ischaemia, however it can be found frequently in acute phase of ischaemic stroke. The authors' aim was to investigate whether the decreased activity of natural anticoagulants is accompanied by worsening of symptoms in ischaemic stroke. PATIENTS AND METHOD: Sixty-eight acute ischaemic stroke patients were investigated. Severity of symptoms were assessed and followed by the NIH Stroke Scale. Antithrombin-Ill, protein C, protein S activities, and concentration of C-reactive protein (CRP) were measured within 48 hours after onset of ischaemic stroke. RESULTS: Progressing stroke was found in 29% of patients. Decreased activity of at least one natural anticoagulant proteins was present in 31% of patients. Progression of stroke symptoms occurred in 76% of patients with decreased natural anticoagulant activity, while this proportion was only 9% in those with normal natural coagulation inhibitor protein activity (p < 0.01). Progressing stroke was also more frequent in patients with elevated CRP value (60%) than in those with normal CRP level (11%; p < 0.05). Decreased activity of natural anticoagulants was more frequent in patients with elevated CRP concentration compared with patients with normal CRP. CONCLUSION: The results demonstrate the importance of decreased activity of natural anticoagulants in acute phase of ischaemic stroke. This abnormality was present in about 1/3 of stroke patients. The decreased activity of natural coagulant inhibitor proteins may play an important role in development of progressing stroke thus indicating unfavourable outcome. PMID- 15884397 TI - Experimental demyelination caused by primary oligodendrocyte dystrophy. Regional distribution of the lesions in the nervous system of mice [corrected]. AB - BACKGROUND AND PURPOSE: Heterogeneity of multiple sclerosis lesions has been recently indicated: In addition to T-cell-mediated or T-cell plus antibody mediated autoimmune mechanisms (patterns I-II) two other patterns (IIl-IV) were described. Patterns Ill-IV are characterized by primary oligodendrocyte dystrophy, reminiscent of virus- or toxin-induced demyelination rather than autoimmunity. It was described more than 30 years ago that dietary application of a copper-chelating agent called cuprizone results in primary oligodendrocyte degeneration which is followed by demyelination. The aim of the present study was to examine the regional distribution of cuprizone induced oligodendrocyte dystrophy and demyelination in the nervous system of mice. MATERIAL A METHODS: Demyelination was induced in male weanling Swiss-Webster mice by feeding them on a diet containing 0.6% (W/W) cuprizone bis(cyclohexanone)-oxalyldihydrazone (G. F. Smith Chemical, Columbus OH) for 8 weeks. Animals were sacrificed after 3, 7, 14, 27, 35, 56 days of cuprizone administration. Samples were taken from corpus callosum, anterior commissure, optic nerve, cervical spinal cord and sciatic nerve. Samples were examined by immunohistochemistry, in situ hybridization for myelin proteins and myelin protein mRNA-s, respectively. Conventional neuropathological stainings and electron microscopy was also performed. RESULTS: Oligodendrocyte degeneration and demyelination followed a particular standard pattern in the central nervous system. Profound myelin loss developed in the superior cerebellar peduncle, anterior commissure and corpus callosum, whereas the optic nerves, velum medullare anterior and spinal cord showed little or no demyelination. Sciatic nerves were unaffected. No infiltration by lymphocytes or blood-brain barrier damage was observed during cuprizone treatment. CONCLUSION: Cuprizone induced oligodendrocyte damage and demyelination follows a particular standard pattern in the central nervous system of mice. Cuprizone induced demyelination might be considered as a model for human demyelinating disorders with primary oligodendrocyte dystrophy and apoptosis. PMID- 15884398 TI - [Clinical features of corticobasal degeneration]. AB - Corticobasal degeneration was described in 1968 by Rebeiz, Kolodny and Richardson, who characterized the disease as a syndrome of asymmetric akinesis and rigidity, dystonia of the upper limb, apraxia, myoclonus and dementia. Atrophy of the frontal and parietal lobe, neuronal loss, gliosis and achromatic neurones (and nowadays astrocytic plaques) are the characteristic pathological features of the disease. Corticobasal degeneration is a rare or a rarely recognized disease and it is frequently misdiagnosed as Parkinson's disease. According to the Lang's criteria, corticobasal degeneration can be diagnosed in the presence of rigidity and one cortical symptom (apraxia, cortical sensory loss, alien hand) or in a patient with rigidity, dystonia and focal reflex myoclonus. Exclusion criteria are early dementia (as in primary degenerative dementias), early vertical gaze problems (as in progressive supranuclear palsy), resting tremor and good, sustained therapeutic response to levodopa (as in Parkinson's disease), severe autonomic problems (as in multiple system atrophy) and any pathology on imaging studies which might explain the clinical symptoms. It should be mentioned, that recently early dementia is recognized as an initial symptom of corticobasal degeneration. The authors present a case and review the literature to call attention to this disorder. PMID- 15884399 TI - [Calpain-3 gene defect causing limb gird muscular dystrophy in a Hungarian family]. AB - Limb gird muscular dystrophies (LGMD2) are a clinically and genetically heterogeneous group of hereditary diseases with autosomal recessive trait, characterized by progressive atrophy and weakness predominantly in the proximal limb muscles. The authors present clinical, histological, immunohistochemical and immunoblot results of two sisters suffering from so far unclassified autosomal recessive limb girdle muscular dystrophy. Haplotype analysis for genes possibly involved in autosomal recessive limb girdle muscular dystrophies was performed in the genetically informative family. All of the results pointed to a molecular genetic defect of the calpain-3 (CAPN3) gene. Direct sequencing of the CAPN3 gene revealed compound heterozygous state for two mutations previously described in association with limb girdle muscular dystrophy, proving pathogenicity. The authors would like to emphasize the importance of the above described combined strategy in diagnosing limb girdle muscular dystrophies. PMID- 15884400 TI - [Headache as a chief subject in the Hungarian medical literature]. AB - BACKGROUND: Less than 1.2% of papers published in the first 50 volumes of the journal Clinical Neuroscience/Ideggyogyaszati Szemle--the major official journal of Hungarian neurologists--focused on headache despite the fact that headache is among the most frequent complaints in neurological consultations. In the current study the authors evaluated the presence of headache as the main topic in articles of the Hungarian medical literature. METHODS: They identified full publications on headache by handsearching all volumes of the journal Clinical Neuroscience/Ideggyogyaszati Szemle from 1950 till the end of 2003. Electronic searches were performed to find Hungarian papers focusing on headache using the bibliographic databases of the Hungarian National Library of Healthcare (Hungarian Medical Bibliography, HMB) and the American National Library of Medicine of the National Institutes of Health (Medline and Oldmedline). Results of handsearch and electronic searches were cross checked for the journal Clinical Neuroscience/Ideggyogyaszati Szemle. RESULTS: Of the 2618 full papers published in Clinical Neuroscience/Ideggyogyaszati Szemle headache was the main topic in 32 articles (1.2%), most of them published after 1985. The electronic search of the HMB resulted in 132 documents in 41 journals, whereas using the PubMed search engine, they identified 66 Hungarian papers on headache. Migraine was the most frequent topic of interest in papers found by all three searches. The HMB search identified all headache articles published after 1990 in Clinical Neuroscience/Ideggyogyaszati Szemle which were identified by handsearch. The Oldmedline, database contained four of the seven papers identified by handsearch from 1954-1964. After 2002, the start of indexing Clinical Neuroscience/Ideggyogyaszati Szemle in the Medline, the only article identified by handsearch was also found by the electronic search. CONCLUSION: After the lack of interest until the mid-1980-ies, headache became a frequent topic in the Hungarian medical literature. Bibliographical data of articles on headache published after 1990 can be identified by electronic searches of the Hungarian and international bibliographic databases using carefully constructed but simple search strategies. An increasing presence of Hungarian headache research was found in international journals in the last two decades. PMID- 15884401 TI - [Psychic consequences on the elderly of sexual abuse in their youth]. AB - Three women, aged 64, 65 and 60 years, who were admitted for psychopathology revealed for the first time that they had been sexually abused as a child by a relative. The first woman sought help following the death of her husband, the second after her daughter was raped, and the third suffered from increasing cognitive impairment. Through therapy, they learned how to process their history of incest. In psychiatric patients, the prevalence of sexual abuse in their youth varies from 5-45% in different studies, depending on the definition of sexual abuse. Two Dutch studies of elderly psychiatric inpatients found a prevalence of 16% sexual abuse in 32 male and female patients and 8% in 110 female patients, respectively. Sexual abuse may have a great, lifelong impact on the victims. Many psychiatric and psychological consequences are found in adult populations, but no study has yet included the elderly. All clinicians should be aware that signs and symptoms in the elderly might be related to sexual abuse. PMID- 15884402 TI - [Pneumococcal vaccination; arguments for informing parents about the options available]. AB - Invasive pneumococcal disease in childhood causes meningitis, sepsis and pneumonia. Current pneumococcal vaccines are estimated to prevent 50% of Dutch pneumococcal meningitis and 40% of pneumococcal sepsis. In 2001, the Health Council of The Netherlands emphasised the importance of universal pneumococcal vaccination of small children. However, the Dutch Minister of Health, Welfare and Sports stated that the vaccine is too expensive for the national vaccination programme (NVP). Child health clinics do not educate parents about vaccines that are not available in the NVP, and therefore parents are not informed about the availability of an effective pneumococcal vaccine. We argue that child health clinic physicians should inform parents about the limitations of the NVP in order to put expectations about the programme into perspective. Educating parents that the NVP is very worthwhile but does not include every possible or available vaccine will strengthen confidence in the NVP. Parents who then want to know which effective vaccines are available should be provided with the information they request. In view of the Health Council recommendations, the pneumococcal vaccine should then be specifically mentioned. PMID- 15884403 TI - [Antithrombotic therapy during percutaneous coronary interventions]. AB - Antithrombotic therapy is essential during percutaneous coronary interventions for the prevention of peri-procedural death and myocardial infarction. The most commonly used agents are aspirin, clopidogrel and heparin in patients treated by percutaneous angioplasty or receiving an arterial stent. Glycoprotein IIb/IIIa receptor antagonists such as abciximab are indicated during percutaneous interventions in high-risk-patients as well as, in principle, in all patients with an acute coronary syndrome with ST-segment elevation undergoing primary percutaneous angioplasty. In patients with so-called drug-eluting stents, clopidogrel should be continued for several months longer than the usual 30 days. PMID- 15884404 TI - [Mortality among non-western migrants in The Netherlands]. AB - Mortality among 10 groups of non-western migrants to The Netherlands, observed in the period 1995-2000, is compared with mortality among people who were born in and whose parents were born in The Netherlands. The migrant groups concerned consisted of people who were born in, or whose parents were born in Turkey, Morocco, Surinam, The Netherlands Antilles, Ghana, Somalia, Iraq, Iran, Afghanistan and Vietnam. Differences in mortality were adjusted for age, marital status, region, degree of urbanization, and socioeconomic status. Despite the fact that most migrants originate from countries with a substantially higher mortality rate than The Netherlands, most groups had similar or more favourable total mortality rates than native Dutch people. Men from Turkey and Surinam had slightly elevated mortality rates and men and women from Somalia had a notably higher mortality rate than native Dutch people. The generally favourable mortality rates among migrants are the result of two compensating phenomena: higher mortality among young migrants than among young native Dutch people, and lower mortality among elderly migrants than among elderly native Dutch people. An analysis of cause-of-death patterns revealed relatively low mortality from cardiovascular diseases, cancer and respiratory diseases in most migrant groups, and relatively high mortality from infectious diseases and injuries. These findings are unlikely to have been influenced by incomplete registration of mortality. Selective migration may play a role--some migrant groups have a relatively high level of education for example. Also some of the findings may be explained by a difference in timing between the health benefits and the health risks of migration. Migrant health could be benefiting from the favourable socioeconomic, public health and health-care conditions in The Netherlands, but not yet be affected by the higher risks of cancer and cardiovascular disease associated with prosperity. PMID- 15884405 TI - [Practice guideline 'Diagnostic techniques for soft tissue tumours and treatment of soft tissue sarcomas (revision)']. AB - Members of the Dutch working group on soft tissue tumours developed an up-to standard evidence-based multidisciplinary clinical practice guideline for the diagnosis of soft tissue tumours and the treatment and follow-up of soft tissue sarcomas, in cooperation with the Dutch Association of Comprehensive Cancer Centres and the Dutch Institute for Healthcare Improvement. A soft tissue sarcoma is defined as every non-epithelial tumour that does not originate in haematopoietic or lymphatic system, central nervous system or bone. The guideline lists 'alarm signals' to raise awareness of malignancy and recommends consulting a multidisciplinary team. Non-invasive imaging has to be completed before proceeding to any invasive (diagnostic) procedure or assessment of dissemination. Aspiration cytology can be useful for differentiating between sarcoma and other malignancies. A definite diagnosis is obtained by means of image-guided needle biopsy. Tumours will be classified according to the World Health Organization and graded according to the Federation Nationale des Centres de Lutte Contre le Cancer. Surgical excision with a tumour free margin of 2 cm is the core of therapy, taking into account vital structures when necessary. In case of small superficial tumours (diameter < or = 3 cm) excision biopsy may be justified. Radiotherapy is almost always necessary and certainly indicated when wide margins are impossible even after re-resection. In the case of primary metastatic disease, an individual decision should be taken after multi-disciplinary consultation concerning the possibility of curative or palliative treatment. Neither neo-adjuvant nor adjuvant chemotherapy is standard. Chemotherapy may be useful in metastatic disease. The guideline advises referring patients who are eligible for chemotherapy to a centre and that they should be included in a study protocol. PMID- 15884406 TI - [From gene to disease; EVC, EVC2, and Ellis-van Creveld syndrome]. AB - Ellis-van Creveld syndrome is an autosomal recessive disorder characterised by short stature with short limbs, postaxial polydactyly and congenital cardiac defects. The syndrome can be caused by mutations in the EVC gene or the EVC2 gene. The genes are located close to each other in a head-to-head configuration on chromosome 4p16. Clinical diagnosis can be confirmed by DNA analysis, which is currently offered by two laboratories in Italy. PMID- 15884407 TI - [The attitude of Dutch women of Turkish ancestry towards measures for the prevention and treatment of vitamin D deficiency; the results of focus group interviews]. AB - OBJECTIVE: To obtain insight into the attitude of Dutch women of Turkish ancestry towards various possible measures for the prevention and treatment of vitamin D deficiency. DESIGN: Qualitative, focus group interviews. METHOD: Four focus group meetings of 4-13 women of Turkish ancestry (mean age: 31.8-47.1 years) were held in community centres in Amsterdam. Discussions were held with the aid of an interpreter in response to questions about exposure to sunlight, vitamin D supplements, fortification of foods with vitamin D, and the consumption of vitamin D rich foods. The discussions were recorded, translated, and transcribed, and the recurring themes were analysed. RESULTS: Participants had a strong preference for vitamins from 'natural sources' as compared with vitamin supplements. Concerns about weight gain and overdose as a result of vitamin supplement use were expressed frequently. Most women only found the use of vitamin D supplements acceptable if prescribed by a physician. For religious/cultural reasons, many women were not willing to alter their dress to expose more skin to sunlight, or were only willing to do so in the strict absence of men that are not members of the family. Participants were reluctant to consume vitamin D-fortified margarine products because of concern about the possible presence of pork fat and the fear that consumption of these products would induce weight gain. CONCLUSION: Among Dutch women of Turkish ancestry, cultural and religious beliefs and concern about the possible side effects can impede the acceptance of advice on how to combat vitamin D deficiency. This should be taken into consideration when giving information on the prevention and treatment of vitamin D deficiency. PMID- 15884408 TI - [Effect of vaccination against pertussis on the incidence of pertussis in The Netherlands, 1996-2003]. AB - OBJECTIVE: To determine whether booster vaccination of 4-year-old children with an acellular pertussis vaccine, which has been included in the national vaccination programme since October 2001, has decreased the incidence of pertussis. DESIGN: Descriptive. METHODS: Surveillance data were studied: mandatory notifications to the Health Inspectorate and reports of hospital admissions from the National Medical Register. RESULTS: During the past 7 years, there has been an increase in the incidence of pertussis every 2-3 years (1996, 1999, 2001). Moreover, the annual incidence in 1996-2003 was higher than in 1989 1995. As in previous years, the yearly peak incidence for hospital admissions due to pertussis was observed among nurslings, especially those younger than 3 months of age. In 2002 compared to 2000, the incidence among 3-4-year-olds on the basis of notifications and hospitalisations was 45% and 62% lower, respectively, very likely due to the booster vaccination for 4-year-olds introduced in 2001. The greatest decrease in the incidence was also observed among the 4-year-olds in 2003. CONCLUSION: Pertussis is still endemic in The Netherlands with a higher incidence than before the epidemic of 1996-1997. Severe disease often occurs, especially among unvaccinated children < 1 year of age. From January 2005 onwards, the vaccinations in the first year of life have been given with an acellular pertussis vaccine. However, since such infants are too young to be protected by vaccination alone, more information is needed on the most important sources of infection of nurslings in The Netherlands. PMID- 15884409 TI - [Missed hyperinsulinaemia in a patient with an insulinoma]. AB - In a 57-year-old man with symptomatic hypoglycaemias which gave cause to suspect an insulinoma, normal insulin levels were initially found. A repeated fasting assay at another hospital did, however, reveal the expected hyperinsulinaemia. Scans revealed an abnormality in the pancreas. After surgical removal of the insulin-producing tumour the patient made a quick recovery. The diagnosis of organic hyperinsulinaemia is established by demonstrating inappropriately high serum-insulin concentrations during fasting hypoglycaemia. The diagnostic normative values are based on the classic polyclonal method of determination. This new highly-specific insulin assay has no cross-reactivity with pro-insulin, which is often produced disproportionately more by an insulinoma. As a result of this false-normal insulin values are found. Therefore new normative values are needed for the newer insulin assays when diagnosing an insulin-producing islet cell tumour. Pro-insulin and C-peptide assays may play a useful role in this. PMID- 15884410 TI - [Allowing a patient in a vegetative state to die in hospital under the direction of a nursing-home physician]. AB - In the case of a 52-year-old man, who was in a vegetative state after resuscitation following an unwitnessed cardiac arrest, the nursing-home physician assumed responsibility for the total treatment in a transfer unit of the hospital. His systematic, multidisciplinary approach enabled direction of the complex situation in which many medical and paramedical personnel were involved. When an airway infection occurred as a complication shortly after responsibility for the patient had been assumed, the nursing-home physician could take a well considered decision to withdraw medical treatment on the basis of the treatment plan which had been formulated in a short time and contact with the family. In clinical practice it is difficult to determine the most appropriate moment to withdraw all medical therapy, including artificial nutrition and hydration, in order to prevent a hopeless vegetative state. This case illustrates how hospital physicians and nursing-home physicians may cooperate during the 'waiting phase' of the clinical course of a vegetative state. A hopeless vegetative state can be prevented by using these transmural possibilities for cooperation, including an early input of knowledge and experience about the long-term course of a vegetative state. PMID- 15884411 TI - [Seizures in foreign newborns due to maternal vitamin-D deficiency]. PMID- 15884412 TI - [Seizures in foreign newborns due to maternal vitamin-D deficiency]. PMID- 15884413 TI - [Seizures in foreign newborns due to maternal vitamin-D deficiency]. PMID- 15884414 TI - [Spirometry in patients with COPD in family practice and in a lung function laboratory equally trust worthy, but not interchangeable]. PMID- 15884415 TI - A second opinion. PMID- 15884416 TI - Low vision standards. PMID- 15884417 TI - Kidneys need some attention. PMID- 15884418 TI - Dynamic visual fields of one-eyed observers. AB - BACKGROUND: The horizontal binocular visual field can extend to more than 200 degrees, while a monocular field is limited to 160 degrees. Additionally, the nose and other facial structures may block the monocular field further during certain eye movements. The purpose of this study was to compare the monocular against the binocular visual field and determine if head and eye movements can functionally overcome any measured deficit. METHODS: In Experiment 1, visual fields were measured monocularly with a bowl perimeter using 5 fixation positions. Binocular visual fields were calculated by combining the monocular visual field with its mirror image. In Experiment 2, subjects were allowed to make head, eye, and body movements to search for flashing lights 360 degrees around them, spaced every 45 degrees. The numbers of lights identified were compared for the subjects performing monocularly versus binocularly. RESULTS: The size of the overall monocular visual field was found to vary between 48% and 76% of the binocular visual field, depending on eye position. For the flashing light experiment, head and eye movements could not overcome the entire visual-field deficit with monocular viewing. Monocular performance remained 11.4% less than binocular performance. CONCLUSIONS: The visual-field deficit seen with monocular viewing is greatest with nasal fixation, and head and eye movements cannot totally compensate for this deficit when viewing time is limited. Vision standards that require full visual fields in each eye are more appropriate for occupations in which peripheral visual targets must be identified and visual search time is limited. PMID- 15884419 TI - Choroidal metastasis: case reports and review of the literature. AB - BACKGROUND: Choroidal metastases are recognized as the most common intraocular malignancy. Their diagnosis has become more common due to increased emphasis on comprehensive eye examinations for cancer patients and the improved life expectancy of patients with metastatic disease. They are most prevalent in female patients with breast cancer and male patients with lung cancer. METHODS: A thorough fundus examination, coupled with the use of A-scan and B-scan ultrasonography, will aid in their diagnosis. There are many ways of treating these tumors, including radiation therapy, chemotherapy, and monitoring. CASE REPORTS: Two case reports of patients diagnosed with choroidal metastases are discussed. Case 1 involved a patient with lung cancer who manifested a large bullous exudative retinal detachment due to an underlying choroidal metastasis. Case 2 dealt with treatment of a patient with metastatic transitional cell cancer who manifested a shallow exudative retinal detachment caused by a choroidal metastasis. CONCLUSIONS: Due to progress of chemotherapeutic medications, the number of patients who manifest choroidal metastases will continue to increase. It is essential for the practitioner to be able to recognize this disease process to prevent visual loss and institute referral for proper treatment for metastatic disease. PMID- 15884420 TI - Leber's hereditary optic neuropathy: a case report. AB - BACKGROUND: Leber's hereditary optic neuropathy (LHON) is a bilateral optic neuropathy of mitochondrial inheritance that produces significant painless, central vision loss and dyschromatopsia. LHON usually occurs in young males between the ages of 15 and 30 years and manifests an episode of subacute or acute vision loss in one eye, with the opposite eye becoming involved weeks to months later. Approximately 80% to 90% of all LHON patients are male. While the disease usually presents itself around the third decade of life, its onset ranges anywhere from 5 to 80 years. CASE REPORT: We report a case of an uncooperative 12 year-old Hispanic boy who was brought to our group practice following referral from an outside optometrist for amblyopia therapy. Following the workup by the binocular vision clinician, a neuro-ophthalmic consultation was obtained, eventually leading to the diagnosis and confirmation of LHON. CONCLUSION: Leber's hereditary optic neuropathy may manifest signs and symptoms that mimic common ophthalmic entities. Teenage males often are reluctant to report its subtle clinical findings, making its discovery even more challenging. LHON should be kept in mind as a possibility for anyone who manifests unexplained visual loss. PMID- 15884421 TI - Sector retinitis pigmentosa. AB - BACKGROUND: Retinitis pigmentosa (RP) is one of the most common hereditary retinal dystrophies and causes of visual impairment affecting all age groups. The reported incidence varies, but is considered to be between 1 in 3,000 to 1 in 7,000. Sector retinitis pigmentosa is an atypical form of RP that is characterized by regionalized areas of bone spicule pigmentation, usually in the inferior quadrants of the retina. CASE REPORT: A 57-year-old Hispanic man with a history of previously diagnosed retinitis pigmentosa came to the clinic with a longstanding symptom of decreased vision at night. Bone spicule pigmentation was found in the nasal and inferior quadrants in each eye. He demonstrated superior and temporal visual-field loss corresponding to the areas of the affected retina. Clinical measurements of visual-field loss, best-corrected visual acuity, and ophthalmoscopic appearance have remained stable during the five years the patient has been followed. DISCUSSION: Sector retinitis pigmentosa is an atypical form of RP that is characterized by bilateral pigmentary retinopathy, usually isolated to the inferior quadrants. The remainder of the retina appears clinically normal, although studies have found functional abnormalities in these areas as well. Sector RP is generally considered a stationary to slowly progressive disease, with subnormal electro-retinogram findings and visual-field defects corresponding to the involved retinal sectors. CONCLUSION: Management of RP is very difficult because there are no proven methods of treatment. Studies have shown 15,000 IU of vitamin A palmitate per day may slow the progression, though this result is controversial. Low vision rehabilitation, long wavelength pass filters, and pedigree counseling remain the mainstay of management. PMID- 15884422 TI - Emergence of a rehabilitation medicine model for low vision service delivery, policy, and funding. AB - BACKGROUND: A rehabilitation medicine model for low vision rehabilitation is emerging. There have been many challenges to reaching consensus on the roles of each discipline (optometry, ophthalmology, occupational therapy, and vision rehabilitation professionals) in the service delivery model and finding a place in the reimbursement system for all the providers. METHODS: The history of low vision, legislation associated with Centers for Medicare and Medicaid Services coverage for vision rehabilitation, and research on the effectiveness of low vision service delivery are reviewed. RESULTS: Vision rehabilitation is now covered by Medicare under Physical Medicine and Rehabilitation codes by some Medicare carriers, yet reimbursement is not available for low vision devices or refraction. Also, the role of vision rehabilitation professionals (rehabilitation teachers, orientation and mobility specialists, and low vision therapists) in the model needs to be determined. In a recent systematic review of the scientific literature on the effectiveness of low vision services contracted by the Agency for Health Care Quality Research, no clinical trials were found. The literature consists primarily of longitudinal case studies, which provide weak support for third-party funding for vision rehabilitative services. CONCLUSIONS: Providers need to reach consensus on medical necessity, treatment plans, and protocols. Research on low vision outcomes is needed to develop an evidence base to guide clinical practice, policy, and funding decisions. PMID- 15884423 TI - Promoting independence through low vision rehabilitation. PMID- 15884424 TI - Strong staff can pay huge dividends. PMID- 15884425 TI - AOA Eye Care Benefits Center carefully watches coding issues. PMID- 15884426 TI - Questions on HIPAA Security Regulation. PMID- 15884427 TI - Marginal donors in kidney transplantation. AB - BACKGROUND: Due to organ shortage, it is not easy to find an ideal donor in renal transplantation. To enhance donor pool, marginal donors can be acceptable even in living renal transplantations. We compared optimal and marginal donor transplantations in terms of graft and patient survival. METHODS: We performed 32 living related renal transplantations, 11 of which were from marginal donors, from January 1999 to December 2000. Recipients from optimal and marginal donors were compared in terms of renal function, acute tubular necrosis (ATN) and acute rejection (AR) rates. RESULTS: Although mean serum creatinine level in the seventh day was found to be higher in the recipients from marginal donors (1,88 mg/dl vs. 2,51 mg/dl); it did not reach to a statistical difference (p = 0.1). Serum creatinine levels after thirtieth day, ATN and AR rates were statistically similar in both groups, as well. In optimal and marginal groups, 3-year graft survival rates were found to be 85 and 72 percent respectively (p = 0.72). 3-year patient survival rates were also found to be similar in both groups (90% vs. 91%) (p = 0.91). CONCLUSION: Functional and survival results in the transplantations from marginal donors were as good as from optimal donors. So, we thought that marginal donors could be used safely with a good preoperative evaluation to increase donor pool. PMID- 15884428 TI - Protocol biopsy of donor kidney in renal transplantation. AB - This review article defines the term marginal (suboptimal) donor of organs and there is a summary of current opinions on the importance of protocol biopsies of kidneys performed in these donors during graft collection. Bioptic examination of kidneys in marginal donors provides information that cannot be obtained in any other way. Most authors engaged in this problem hold the view that evaluation of the extent of involution changes and the decision about the appropriateness of the organ for transplantation cannot be based only on the age criteria and there should always be a biopsy performed in suboptimal donors. Histological examination enables an objective evaluation of a pre-existing morphological lesion; especially the extent of glomerulosclerosis, interstitial fibrosis and vascular damage that may limit the functional level and vitality of the graft. As marginal are usually considered kidneys with more than 20% of glomerulosclerosis, kidneys with interstitial fibrosis affecting more than 25% of cortex or kidneys with a finding of arteriosclerotic changes causing more than 25% obliteration of the vascular lumen. It has to be considered, during a transplantation of a marginal kidney, that a long-term survival of the graft will be significantly decreased. But because the number of patients waiting for transplantation is increasing faster than the number of transplants performed, there are possibilities mentioned briefly in this article advising how to optimally use these marginal kidneys as well. PMID- 15884429 TI - Reconstructive surgery of the extensive ureter strictures using human dura mater allografts. AB - OBJECTIVE: To present and evaluate our experiences with use of human dura mater allografts in reconstructive surgery of long ureter strictures; first human derived, collagen based, oligocellular biomaterial utilized in genitor-urinary reconstructive surgery. To describe on the basis of our experiences with dura mater preferable from a technical and biological standpoint features of biomaterial as a matrix for the ureter regeneration in this condition. We also assessed a technical aspects, suitability and efficacy of the new operative method. PATIENTS AND METHODS: Between 1980 and 1992, in our search for ideal biomaterial useful for reconstructive surgery of extensive ureter obstruction we used to apply human dura mater allografts. A total of 6 females and 2 males were treated with reconstructive surgery with human dura mater allografts utilized for supplementation of the ureter wall defect. Diagnosis was based on ultrasonography, excretory urography and retrograde ureteropyelography. Imaging studies revealed obstructed ureter segment of at least 4 cm length. RESULTS: In all cases procedure was completed without any complications. Hospitalization after the surgery lasted approximately 8-10 days. Early and late follow-up excretory urography demonstrated lack of obstruction in the operated ureter segment. Long term follow-up of 12 months to 18 years (meanly 8,75 years) showed no signs of renal function deterioration, without urine obstruction on the operated side in all patient. Fluoroscopy scans showed signs of peristaltic wave in the operated ureter segment. CONCLUSIONS: Both a supplementary biomaterial used and a new operative method proved to be a promising option in reconstruction of long ureter strictures. Unfortunately a threat of prion related diseases, which resulted in exclusion dura mater grafts and all biomaterial originated from nervous system from transplantology, forced us to search for new suitable material. PMID- 15884430 TI - Reconstruction of long ureteral obstructions using xenogenic acellular collagen membranes. AB - OBJECTIVE: Extensive stricture of ureter still poses a difficult challenge for urologic surgons, especially when its middle part is affected. We present a reconstructive method of long ureteral strictures with use of xenogenic collagen membranes as a scaffolding to promote new tissue growth in the area of extensive ureter defect. PATIENTS AND METHODS: Between 1980 and 2002 we applied and evaluated results of the new procedure in a total of 7 females and 3 males suffering from a various degree of hydronephrosis caused by extensive ureter stricture. RESULTS: Early and long-term follow-up excretory urography demonstrated lack of obstruction in the operated ureter segment in all cases. CONCLUSIONS: Easy surgical procedure, no complications after surgery excellent short and long-term results prove the efficacy and usefulness of the presented therapeutic option. PMID- 15884431 TI - Reconstructive surgery of male urethra using human amnion membranes (grafts)- first announcement. AB - OBJECTIVE: Assessment of technical aspects of the surgical procedure and usefulness, suitability and efficacy of human amnion grafts as a biomaterial in reconstructive surgery of strictured male urethra. PATIENTS AND METHODS: Human amnion membranes were used in 2 men suffering from long, recurring strictures of urethra. Narrowed part of urethra was careful prepared and cleaned of adjacent tissue. Then a longitudinal incision was performed through the whole length of strictured segment and then it was covered with human amnion membrane. RESULTS: Hospitalization time 4 to 5 days. The Foley catheter was removed 2 weeks after surgery 3 months after surgery controlled urethrographies and urethroscopies show wide urethra lumen, wider than in adjacent parts. In urethroscopy operated place covered with epithelium, smooth, without scare. CONCLUSIONS: Human amnion grafts and described technique seem to be a promising method of managing long, recurring male urethra strictures but need long-term follow-up and analysis of more cases. PMID- 15884432 TI - Cells in the transplanted organ live in the recipient their own life. PMID- 15884433 TI - Biological effects of bone marrow in transplanted limb--a review. AB - Transplantation of a vascularized limb should be considered as a double graft: composite parenchymatous non-lymphoid tissue (skin, subcutaneous tissue, muscles, nerves and bones in a block) and bone marrow (BM) tissue. Ready-to-function BM releases mature and precursor cells migrating to the recipient tissues immediately after transplantation. Donor BM-derived cells home to recipient BM cavities and lymphoid organs (LO) and to lesser extent to the non-lymphoid tissues. Interestingly, no acute and subacute graft-versus-host reaction develops. Cellular microchimerism is detected with the presence of donor cells in recipient tissues, at least as long as the transplanted limb is not rejected. Free donor DNA appears in high concentrations in recipient tissues at the time of rejection. The biological significance of the cellular and DNA microchimerism is not clear. Slight prolongation of limb and free-flap skin grafts survival time may be attributed to microchimerism. The beneficial and unwanted effects of transplanted BM tissue should be taken into consideration in the clinical limb transplantation programme. Long-term experimental and clinical observations will allow to draw more clinically applicable conclusions. Abbreviations used in this review: BM: bone marrow, BMC: bone marrow cells, BMTx: bone marrow transplantation, il-BMTx: in limb-bone marrow transplantation, iv-BMCTx: intravenous isolated bone marrow cell transplantation, MLN: mesenteric lymph node, SPL: spleen, LO: lymphoid organs, PHA: phytohemagglutinin, ConA: concavalin A, PWM: pokeweed mitogen, CsA: cyclosporin A, TBI: total body irradiation. PMID- 15884434 TI - Stem cells of the human skin epithelium--can they be isolated and resume function as single-cells transplanted into recipient skin defects? AB - A review, supplemented by own observations, on the human epidermal stem cells on identification, anatomical location, proliferation capacity and storage of these cells has been presented. There are no specific markers for epidermal stem cells and the criteria for identification are based on low kinetics of cycling, retention of bromdeoxyuridine or thymidine, expression of beta integrin, transferrin receptor, protooncogene p63 and adherence to collagen IV. The regenerative capacity resides not only in stem cells but also in their more committed progeny. Problems concerning practical aspects as transplantation of epidermal stem cells at single-cell level or in context of whole epidermis for covering large skin defects and in gene therapy are discussed. PMID- 15884435 TI - Human skin preserved in anhydric sodium chloride for months can be successfully transplanted. AB - Human skin can be preserved in pulverized sodium chloride dehydrated at 240C for 2 hours at room temperature for periods of weeks or months and successfully transplanted to scid mouse, retaining its normal morphological structure. Fragment of skin of a size of 10 x 10 x 6 mm were harvested during elective vascular and orthopaedic surgery of lower limbs, dried of blood and placed in anhydric sodium chloride powder in tight sealed containers. Prior to transplantation to scid mice, the specimens were desalinated and rehydrated. Specimes preserved for 1 to 6 months and harvested 3-4 weeks after transplantation revealed intensive incorporation of bromdeoxyuridine (BrdU) into basal keratinocytes. They expressed p63 and CD29 (stem cells, and transient cells antigens), PCNA (proliferating cell nuclear antigen) and cytokeratin 16 specific for proliferating keratinocytes. Dermal fibroblasts and few large HLA II cells showed a normal structure. Bacterial flora of skin did not change after grafting. We conclude that human skin can survive in a dehydrated state in sodium chloride for months and after transplantation the epidermal basal layer cells give rise to keratinocyte progenies. Skin fibroblasts and some resident immune cells can also survive. PMID- 15884436 TI - Successful transplantation of hepatocytes requires temporary elimination of scavenger and NK cells, partial hepatectomy and ligation of bile duct. AB - Isolated syngeneic and allogeneic hepatocytes transplanted to spleen parenchyma are rapidly disintegrated. Host scavenger monocytes and macrophages are responsible for this reaction. We designed a method overcoming early disintegration of the intra-splenic grafted hepatocytes. It consisted of administration of anti-asialoGM I antiserum eliminating NK cells, sublethal whole body irradiation for depletion of scavenging cells and reconstitution with syngeneic bone marrow cells, ligation of host bile duct, intrasplenic hepatocyte transplantation and 3 consecutive partial hepatectomies. Six months after transplantation a glycogen-rich, trabeculae-forming, dividing hepatocytes, situated along strands of newly-formed fibrous tissue and numerous dilated blind bile canaliculae were observed. There was evidently more bile canaliculae in hosts with ligated bile duct than non-ligated controls. This is the first study showing fibrous tissue formed at the site of hepatocyte implantation and stellate cells are presumably responsible for this process. PMID- 15884437 TI - Repopulation of the heart graft with recipient immature dendritic cells prior to transplantation does not prolong its survival. AB - Experimental studies on allogeneic transplantation have shown that recipient dendritic cells (DC) play a role in peripheral tolerance as well as in rejection of allografts. It is not known whether DC exert their tolerogenic function in the graft or in recipient lymphoid tissue. To answer this question we created a chimeric heart model deprived of its own DC and repopulated by recipient DC. The rationale for this model was to observe whether recipient mature and immature DC located in the graft attenuate recruitment and stimulation of recipient lymphocytes, subsequently prolonging graft survival. Vascularized bone marrow transplants (VBMTx) from the prospective recipient to the lethally irradiated heart donor, which function for a period of 14 days, were used to replace donor DC with prospective recipient either mature or immature DC. Replacement of the donor heart with either of these cellsdid not prolong graft survival. The intra graft microchimerism did not mitigate the allogeneic rejection reaction. PMID- 15884438 TI - Spleen dendritic cells of recipients of allogeneic but not syngeneic heart grafts internalize donor DNA fragments. AB - Microchimerism after allogeneic organ transplantation has been widely documented using DNA identification techniques. However, the question as to whether the detected donor DNA is present in the surviving donor cells, recipient macrophages phagocytizing rejected donor cells or recipient dendritic cells internalizing donor apoptotic bodies or cell fragments has not been answered. We provide evidence that allogeneic organ transplantation is followed not only by cellular microchimerism caused by release of graft passenger cells but also dissemination of donor DNA from the rejecting graft cells and its internalization in recipient dendritic cells (DC). Identification of allogeneic donor DNA was based on detection of the male Sry-PCR product in extracts of DNA from recipient tissues, dendritic cells and their nuclei. Most interestingly, donor DNA could be detected at high concentration in all recipient tissues at the time of rejection. Search for specific localization of allogeneic donor Sry fragments in recipient cells revealed its presence preferrentially in the DC. No donor Sry fragments were detected in recipient DC after syngeneic transplantation. Detection of allogeneic but not syngeneic donor Sry in DC nuclei further strengthens our concept that DC specifically incorporate allogeneic donor DNA fragments. The mechanism of this process requires further studies. PMID- 15884439 TI - Influence of bacterial antigens on activation of human splenic dendritic cells. AB - The dendritic cells (DC) play crucial role in initiation and modulation of immune response especially innate immune response. Toll like receptors (TLR) on DC are receptors involved in innate immunity and recognizing conserved bacterial antigens like LPS and bacterial DNA. TLRs can also respond to some endogenous ligands (heat-shock proteins, heparan sulfate, fibrinogen and the contest of necrotic cells). Recognition of such endogenous substances would be a critical step in response to viruses, tumors and possibly to transplants. We investigated the influence bacterial antigens on splenic cell population enriched in DCs. After incubation with bacterial antigens the percentage of DC expressing HLA-DR+ and CD 123 + cells increased whereas that of CD68+ and CD14+ decreased. In the untreated population of human splenic DC minimal expression of TLR2, TLR3 and CD123 was found, while other receptors were not detected. After incubation with bacteria a marked increase of CD83, TLR2, TLR3 and TLR4 was observed. Treatment with LPS increased expression of TLR2, TLR4, Hsp60 and Hsp90. Stimulation by bacterial DNA resulted mainly in Hsp60 and TLR9 expression. These observation may throw light on the mechanism of exacerbation of the rejection of transplanted organs by microbial stimulation. PMID- 15884440 TI - The reaction of the regional lymph nodes on the skin bacterial and allogeneic antigens. AB - Transplantation of hands in humans has become an accepted therapeutic modality. A hand transplant is composed of various tissues of different degree of immunogenicity. In addition, skin contains own resident bacterial flora that may become virulent in the ischemic and rejecting graft. Discrimination between skin rejection and bacterial inflammation is difficult. The aim of our experimental study was to investigate the cellular reaction to skin bacteria and alloantigen in the regional lymph nodes draining skin and analyse the changes in cell phenotypes. The study was carried out on rats inoculated into hind-limb paw either with S. epidermidis or allogeneic peripheral blood mononuclear cells. An increase in lymph node weight and cell concentration was observed after bacterial infection. After stimulation with allogeneic cells, node mass increased significantly but its cell number rose much less. The percentage of lymph node W3/13 (leukocytes, T cells), W3/25 (CD4), OX8 (CD8), OX6 (class II), OX12 (B cells), EDI (CD14), CD31 (lymphocytes), OX7 (stem cells), and OX62 (migrating dendritic) cells did not change in both groups compared to controls. There was a significant rise of the CD54 (ICAM I) subset after bacterial infection and increase in percentage of OX8-cytotoxic and decrease of W3/25 (helper) subset after allogeneic stimulation. Lack of major differences between stimulated and control contralateral nodes may be explained by systemic reaction to the tested antigens affecting also nodes on the non-injected side. Comparison of the reaction of bacteria and alloantigen stimulated nodes revealed an increase in percentage of OX6, OX12, CD31, CD54, OX33 and OX62 after infection with S. epidermidis, whereas allostimulation brought about only rise in T (W3/13) cell population. Neither stimulation caused increase in expression of class II antigens on T cells. The obtained results demonstrate evident differences in type of response to bacteria and alloantigens. To what extent can bacterial stimulation enhance allogeneic reaction is now under study. Our data are helpful in understanding differences in the character and kinetics of reaction to both types of antigens and may taken into consideration when planning therapy in recipients of hand allografts, immunosuppressive drugs or antibiotics. PMID- 15884441 TI - [Sexual and household violence against adult women and girls]. PMID- 15884442 TI - [Sexual aggression against girls and adult women --definitions and epidemiology]. AB - Various definitions of sexual aggression lead to different rates of incidence and prevalence. Crime statistics indicate the rate of declared judiciable acts (Hellfeld). Research definitions are broader with respect to behavior of the perpetrator, damage to the victim etc. (Dunkelfeld). Taking various data sources into account it emerges that 12-20% of women become victims of sexual aggression, which means that sexual aggression is ubiquitous in society reaching all ages and social classes, women being by far more often exposed than men. PMID- 15884443 TI - [Sexual aggression against girls and adult women--causes and consequences]. AB - The causes of sexual aggression are multiple. Bioevolutionary rooted behavioral dispositions (sexual aggressiveness of men as a reproductive strategy) are enforced by specific male dominated structures of society and socially determined stereotypes of female roles which deter women from self determination of their sexuality; these macrosocial factors combine with individual conditions of socialization, in which violence plays an important role. With respect to the consequences of sexual aggression primary victimization (the trauma itself with physical and psychological harm) can be distinguished from secondary victimization (devaluation and mistrust in the social environment). The degree of negative health outcomes depends on risk factors like previous abuse, severity of the violent act, and lack of social support. Prevention must aim at changes of societal conditions that enhance sexual aggression and the establishment of programs for men and women at risk in which techniques of control of sexual aggression, attenuation and inhibition can be learnt. PMID- 15884444 TI - [Immediate care for women after sexual and physical assault]. AB - The management of recent sexual and physical assault in women has two aims: to provide medical and psychological care for the victim and to prevent possible health consequences of the trauma on one hand, and to facilitate the forensic assessment with regard to the perpetrator's prosecution on the other. The victim should always be encouraged to report an offence to the police and forensic medicine should be called in to assist in the victim's examination, whenever possible. In addition, emergency contraception and prophylactic measures against sexually transmitted infections (STI), especially HIV, must be offered. The profound emotional impact of sexual and physical assault requires knowledgeable and sensitive crisis intervention on the part of the counsellor and referral for ongoing counselling and support must be made available. A well trained and competent sexual assault care team with a sympathetic and non-judgemental attitude can contribute profoundly to the assaulted woman's regaining control over her life and being able to overcome the experienced trauma. PMID- 15884445 TI - [Sexual abuse of female children and adolescents--detection, examination and primary care]. AB - Epidemiological studies show a prevalence of sexual abuse experience among girls from 14-33%. Although indicators of abuse are unspecific, the combination of several findings may be indicative: Somatic signs may be sexually transmitted diseases, vulvovaginal complaints. Psychosocial nonsexual indicators are abrupt behavioural changes, running away from home, eating disorders. Psychosexual signs are hypersexualisation of the language and behaviour, disturbed body image and gender identity. Indirect evidence of abuse is given not only in cases of old vaginal and anal lesions but also in situations, where deep tears of the hymen in the typical localization at the posterior part can be found. The workup and care for children in whom there is suspicion of abuse but no clear evidence asks for highly competent professionals in a multidisciplinary cooperation including pediatric gynecologists, child psychiatrists, children-protection groups and other specialists to avoid on one hand unjustified destabilisation or even destruction of familial structures but to assure on the other hand, that the child victims are treated and followed after in a short and long term comprehensive medical and psychosocial care. PMID- 15884446 TI - [Violence against women in the context of migration]. AB - Since the eighties the percentage of migrant women is increasing continually in Switzerland and presents a new challenge for the public health system. The article will point out specific aspects of migration which are particularly associated with violence against women, besides domestic and sexual violence in their marriages. A successful communication serves as a basis for a reasonable diagnosis, therapy and counselling in health services. Beyond overcoming language barriers, it is essential to consider the physician-patient interaction influenced by the social status and cultural background of each. By using two vignettes specific difficulties in medical encounters with migrant women will be demonstrated and possible interventionsfor clinical practice will be offered complemented by references on existing services in Switzerland. By an interdisciplinary co-operation of physicians, social workers, mediators, lawyers and anthropologists a professional care in face of these complex issues will be guaranteed. PMID- 15884447 TI - [Therapeutic concepts and therapy development in the framework of longterm treatment of victims of sexual and domestic violence]. AB - Domestic violence differs from the nonrecurring trauma e.g. in the context of rape by a stranger in different aspects, emphasising the complexity of the victim's reaction and the enmeshment of the perpetrator The violence takes place in an established system of an intimate relationship which the woman was contracting voluntarily. Violence happens repeatedly and often several individuals (partner, children are directly and indirectly involved, which abide a relationship with the perpetrator Therefore the treatment of the posttraumatic stress disorder outstands in several aspects. The victims initially shows difficulties in talking about the experienced violence and are afraid to become unloyal; a therapeutic approach is often possible only after a fairly long time. Most authors agree that trauma therap the three phases (1 security and protection, psychoeducation and stabilisation, (2 trauma confrontation and (3 reintegration. Establishing a secure environment and intrapsychic stabilisation are a requirement for the therapeutic confrontation with the traumata in order to avoid re-traumatisation. Working with domestic violence victims is complex and demanding, experience in the work with trauma victims and regular supervision are fundamental. PMID- 15884448 TI - [The role of governmental and non-governmental institutions in fighting domestic violence against women, with a special focus on prevention]. AB - Domestic violence is the most important violation world wide of human rights of women and one of the most frequent causes for death! The WHO estimates that one of five women experience domestic violence throughout their lifetime. In Switzerland, as much as elsewhere. The batterers and the victims are to be found in all social strata, ages and origins. The costs cause enormous economic damage. Thanks to efficient cooperation of all the governmental and non-governmental institutions victims can be protected a lot better and batterers can be prosecuted. Coordinating the different measures to be taken by all these actors is the central duty of the cantonal intervention projects against domestic violence. Research - in the medical domain, too - is very important. New laws (e.g. domestic violence as officially prosecuted/ expulsion of the perpetrator) should have a preventive effect and express the very clear message: Domestic violence is not a private issue (any longer)! PMID- 15884449 TI - [Examination of victims of sexual violence--the problem of false reports]. AB - Some people claim to be victims of sexual violence and abuse the judicial system by filing a false police report. Generally, motivation for such behaviour is assumed to be that the self-proclaimed victims try to avoid taking responsibility for a sexual act in which they have been an active, willing participant. Concise motivations include attempts to mask a sexual affair from a partner, or to mask a first sexual intercourse by pretending to be the victim of an act of violence. By the nature of many judicial systems that are abused for such purposes, clinical forensic specialists are confronted with such cases far more often than clinical doctors not serving judicial authorities. In adults, history, injuries and other findings often show patterns that can be recognized as evidence of deception or as signs for self-inflicted injury for the trained specialist. This is far less often the case in infants and children. PMID- 15884450 TI - Health insurance as an integral component of health maintenance organization (HMO) urgent need for paradigm shift. PMID- 15884451 TI - Presenile dementia--etiology, clinical profile and treatment response at four month follow up. AB - Dementia is the development of multiple cognitive deficits that includes memory impairment and at least one of the following--Aphasia, apraxia, agnosia or disturbances in executive functioning. The common causes of dementia among the elderly are Alzheimer's disease, vascular dementia, mixed dementia and Lewy body disease. The concept of reversible dementia was introduced in 1980 when a task force sponsored by National Institute of Ageing found 10-12% of dementia cases in older group to have reversible causes such as metabolic-nutritional, drugs, infections, psychiatric disorders etc. In our series of 76 patients in the presenile age group (<65 years), 34.21% (26/76) had a reversible condition underlying the dementia. 43.42% (33/76) had vascular dementia, 13.15% (10/76) had Alzheimer's disease and 9.21% (7/76) had mixed dementia. Hypertension, hyperlipidemia and diabetes mellitus were commoner in the vascular dementia group as compared to the Alzheimer's group. Evaluation of MRI as a tool in diagnosis of dementia showed increased sensitivity of MRI towards detecting lacunes. The potentially reversible dementias comprised infections 14.47% (11/76), metabolic nutritional 14.47% (11/76) and autoimmune diseases 3.94% (3/76). These were characterized by a subcortical dementia. Four month follow up of MMSE in this group showed significant and sustained improvement in the metabolic nutritional group. PMID- 15884452 TI - Effect of body temperature on mortality of acute stroke. AB - OBJECTIVES: The exact relation between body temperature and mortality of acute stroke victims is poorly understood. However, body temperature can affect the outcome of stroke cases in relation to mortality. In the initial (4-12 hrs) hours of stroke, the temperature rise is often neurogenic in origin and mortality is also higher with raised body temperature. Though the anatomical lesion or nature of lesion along with other established risk factors influence the mortality of stroke cases, change in temperature does affect the outcome of stroke cases. We sought to find out the significance of stroke victim's body temperature (within 4 12 hrs) with respect to short-term mortality rate in our study. METHODS: We selected 100 ischemic and 100 hemorrhagic stroke patients proved by CT scan of brain and 2-hrly oral temperature was strictly recorded for the initial 4-12 hrs of admission. RESULTS: It was evident from the study that hyperthermia (>37.5 degrees C) was associated with highest mortality rate in both hemorrhagic (51.78%) and ischemic (56.66%) cases. In normothermic group, mortality was 13.5% and 8.8% in hemorrhagic and ischemic group respectively, whereas hypothermia (<36.5 degrees C) was associated with 0 % mortality. CONCLUSION: Hyperthermia in acute stroke victim carries a bad prognostic parameter in short term basis. PMID- 15884453 TI - Ultrasonographic evaluation of gallbladder volume in diabetics. AB - AIMS AND OBJECTIVE: Ultrasonographic determination of gallbladder volume in diabetics [both type I and type 2], it's comparison with a control group, and correlation of gallbladder volume in diabetics with parameters such as age, sex, body mass index, parity, hyperlipidaemia, and autonomic neuropathy. MATERIALS AND METHOD: Ninety one cases of diabetes mellitus and 40 healthy controls were recruited for the study. A detailed history and physical examination were recorded. Laboratory investigations done were--fasting and postprandial blood sugar, glycosylated haemoglobin, microalbuminuria, and serum lipid profile. Autonomic neuropathy was determined by using simple non-invasive bedside tests. Fasting gallbladder volume was measured by ultrasonography (calculated by ellipsoid formula). RESULTS: The mean fasting gallbladder volume was 18.20 +/- 2.54 ml in type I diabetics and 25.87 +/- 13.90 ml in type 2 diabetics, with a minimum value of 9.30 ml and maximum value of 88 ml. When type 2 diabetics were subgrouped according to the presence of autonomic neuropathy, higher gallbladder volumes were seen in patients with autonomic neuropathy. CONCLUSIONS: Cholecystomegaly, to a significant degree, was found in type 2 diabetics in the present study. It was significantly correlated with age, body mass index, and the severity of autonomic neuropathy. In male type 2 diabetics, gallbladder volume was significantly correlated with LDL cholesterol levels. In female type 2 diabetics, gallbladder volume was significantly correlated with waist-hip ratio. Gallbladder volume also had significant correlation with proliferative diabetic retinopathy, but not with glycaemic control, microalbuminuria, hypertension, or the duration of diabetes. PMID- 15884454 TI - Role of endothelial cell dysfunction in essential hypertension. AB - BACKGROUND: Hypertension is associated with functional and morphological alterations of the endothelium, which disturbs delicate balance of endothelium derived factors resulting in endothelial dysfunction. The endothelial dysfunction could then facilitate the maintenance of elevated peripheral resistance, which would favor the occurrence of atherosclerosis. AIMS AND OBJECTIVES: The aim of the present study was to determine the circulating levels of vasodilators [nitric oxide (NO) and prostacyclin (PGI2)] and vasoconstrictors [endothelin I (ET-I) and thromboxane (TX)A2)], which reflect endothelial cell dysfunction. METHOD: Nitric oxide as nitrites and nitrates (NOx) were measured spectrophotometrically; ET-I, TXA2 (as TXB2) and PGI2 (as 6 keto PGFIalpha) were measured using enzyme immunoassay methods in 54 male subjects having predominantly untreated, mild hypertension and compared with age-matched 75 healthy controls. RESULTS: Significantly higher levels of ET-I (p<0.001) and TXB2 (p<0.001) were found in essential hypertension subjects (EHT) as compared to controls. No significant difference was observed in NOx and 6 keto PGFIalpha between the two groups. There was significant increase (p = 0.005) in the ratio of TXB2/6 keto PGFIalpha in EHT subjects as compared to controls. CONCLUSIONS: Elevated levels of vasoconstrictors in untreated essential hypertension subjects as compared to controls confirmed the presence of endothelial dysfunction, even in mild cases of hypertension. Early detection of endothelial dysfunction may be a useful measure to guide therapy before the damaging effects of hypertension manifests. PMID- 15884455 TI - Advances in bronchoscopy--new and upcoming bronchoscopic methods at the dawn of the 21st century. AB - The last decade has seen the increasing application of bronchoscopic methods such as transbronchial needle aspiration in the staging of lung cancer as well as in the diagnosis of peripheral lung cancer, balloon dilatation using flexible bronchoscopy, development of new tracheobronchial stents to manage central airway obstruction, autofluorescence bronchoscopy for the early diagnosis of lung cancer and pediatric flexible bronchoscopy. There is also a better understanding in the mechanisms and management of hypoxemia during bronchoscopy such as upper airway obstruction. Recent developments include direct endobronchial ultrasound to increase the yield of transbronchial needle aspiration, high magnification bronchoscopy to assist in the diagnosis of early lung cancer and bronchoscopic lung volume reduction to gain the effects lung volume reduction surgery without the need for surgery in patients with severe emphysema. In this article the new and upcoming bronchoscopic techniques are discussed, which we believe will find a broader clinical application in the near future to manage patients in a better way. PMID- 15884456 TI - Mixed infection with three intestinal coccidian parasites in an AIDS patient. PMID- 15884457 TI - Tuberous sclerosis with multisystem manifestations. PMID- 15884458 TI - Waldenstrom's macroglobulinemia. AB - Waldenstrom's macroglobulinemia is an uncommon lymphoplasmacytic lymphoma presenting with hyperviscocity and autoimmune phenomenon. Disease is characterized by bone marrow infiltration by lymphoplasmacytic cells and raised IgM. Bone marrow morphology and immunohistochemistry is important for diagnosis. Course is indolent and anemia and age are most important prognostic factors. Treatment options include alkylating agents, anti-purine anti-metabolites, which though not curative but offer valuable responses. Newer agents like Rituximab and autologous transplant are being tried. PMID- 15884459 TI - Management of acute myocardial infarction--primary angioplasty the treatment of choice! AB - The management goal of acute myocardial infarction is prompt revascularization. Recent years have seen improvements in both pharmacological and mechanical methods of revascularization of the infarct related arteries. This brief review summarises these developments and ongoing efforts. PMID- 15884460 TI - Bilateral internuclear ophthalmoplegia following head injury. AB - A 40-year-old female after a closed head injury presented with bilateral internuclear ophthalmoplegia. Attempted convergence was abnormal and MRI revealed the focal hemorrhage in the medial longitudinal fasciculus region by showing bright signal in the a pontomesencephalic region in the midline on both T1 and T2 weighted images. The patient regained normal ocular mobility after six months of the injury. The medial longitudinal fasciculus, which is believed to be lesioned in cases of internuclear ophthalmoplegia, is an unusual and rare finding, particularly in patients victims of head injury without further neurological signs. Isolated internuclear ophthalmoplegia should be considered in the differential diagnosis when one encounters an adduction deficit in a patient suffering head injury. PMID- 15884461 TI - ARDS in a case of vesicular mole with secondary hyperthyroidism. AB - Vesicular mole is best regarded as a benign neoplasia of the chorion with malignant potentials. In India, the prevalence is one in four hundred pregnancies. We present one such case who presented with thyrotoxicosis and pulmonary edema. PMID- 15884462 TI - Diffuse interstitial skeletal hyperostosis (DISH) in type 2 diabetes. AB - A 58 years type 2 diabetic woman, school teacher by profession, presented with backache, neck pain and generalised weakness since last few months. Pain was mild with stiffness and neck pain was particularly associated with extension of the neck towards back. There was no focal neurological deficit on central nervous system examination. X-ray of lumbo- sacral spine showed prolific osteophytes and new bone formation in the body of lumbar vertebrae. Cervical X-ray showed 'Melting candle-wax' appearance at the anterior to the cervical vertebrae. In view of clinical and radiological association the case was diagnosed as DISH syndrome. It is being presented for its rarity. PMID- 15884463 TI - Polyneuropathy with osteosclerotic myeloma--POEMS syndrome. AB - A 55-years-old male, who presented with insidious onset gradually progressive sensorimotor polyneuropathy, POEMS-syndrome was diagnosed based on polyneuropathy, splenomegaly, hypothyroidism, the presence of IgG-monoclonal serum protein with osteosclerotic lesions and hyperpigmention of skin. Biopsy of the osteosclerotic lesion from the right superior pubic rami was consistent with plasmocytoma. Electrophysiological studies revealed demyelinating sensorimotor neuropathy and biopsy from sural nerve showed demyelinating neuropathy with secondary axonopathy. The patient showed improvement with radiotherapy. This is a rare systemic disease from the clinical spectrum of plasma cell dyscrasias with polyneuropathy. The importance of POEMS syndrome in the differential diagnosis of polyneuropathies has been emphasized. PMID- 15884464 TI - Polyglandular autoimmune endocrinopathy in type 2 diabetes. AB - Polyglandular autoimmunity (PGA) type 2 presenting in childhood is extremely rare. We report a case of type 2 PGA who had hypothyroidism, followed by diabetic ketoacidosis and was later diagnosed to have adrenal insufficiency also. PMID- 15884465 TI - Neurofibromatosis I with unusual hypopigmentation masquerading as leprosy. AB - A case of Neurofibromatosis I (NFI) occurring in association with symmetrical peripheral nerve enlargement and multiple hypopigmented macules strikingly limited to the neurofibromas, with normal to minimally reduced sensations, evoking a strong clinical suspicion of co-existent lepromatous leprosy, is being reported. Leprosy was ruled out by microbiological, histopathological and electrophysiological studies. The case is interesting in view of the hypopigmented macules overlying the neurofibromas, which is an unreported feature of NFI. PMID- 15884466 TI - Papillary carcinoma of thyroid presenting as posterior mediastinal mass with superior vena cava syndrome. AB - Enlarged thyroid with retrosternal extension presenting as anterior mediastinal mass is known. Superior vena cava (SVC) syndrome due to direct invasion from a primary thyroid malignancy is a rare phenomenon. We present a unique case of papillary carcinoma of thyroid extending into the posterior mediastinum with superior vena cava syndrome along with internal jugular and azygous vein thrombosis. PMID- 15884467 TI - Medical philately (medical themes on stamps). Karl Landsteiner (1868-1943). PMID- 15884468 TI - Serum cholesterol in cirrhosis of liver. PMID- 15884469 TI - Dandy-Walker's variant adulthood presentation. PMID- 15884470 TI - Thrombocytopenia and TNF-alpha levels in malaria. PMID- 15884471 TI - Cost effective treatment of acute, uncomplicated Plasmodium falciparum malaria. PMID- 15884472 TI - Epidemic of leptospirosis: an ICU experience. PMID- 15884473 TI - What really works in clinical practice? Becoming more evidence based. PMID- 15884474 TI - Herbal therapies. PMID- 15884475 TI - Autism: understanding conceptual processing deficits. AB - Autism is a behavioral diagnosis characterized by a triad of impairment, which includes impaired communication, impaired social skills, and impaired spontaneity. The outward behavior exhibited by people with autism represents a different cognitive processing style than that seen in typical development. Understanding the way people with autism process and store information is important for intervention. The majority of people diagnosed with autism do not have symptoms of mental retardation and need access to mainstream services to work on the problems presented by daily life. PMID- 15884476 TI - Cryotherapy for treatment of ECT-induced headache. AB - Because headache is a common side effect of electroconvulsive therapy (ECT), this study sought to determine the effectiveness of cryotherapy (i.e., a frozen gel band) in relieving pain in patients with post-ECT headaches, and whether headache intensity and physiological measurements could predict use of an alternative analgesic (rescue medication). We used a quasi-experimental, crossover design to collect data from 31 patients ages 24 to 85 who had been referred for ECT at two medical facilities in San Diego, California. Measurements of patients' pain intensity were made at three intervals: upon perceiving headache, and at 30 and 60 minutes following the cryotherapy or acetaminophen interventions, based on the order of the crossover design. Data were analyzed using Hotelling's T2 and logistic regression. No significant difference was found between cryotherapy and acetaminophen in relieving ECT-induced headaches (p = .420). There was no influence due to the crossover design (p = .313), nor where there significant changes in physiological measures from treatment (p = .420). Logistic regression showed that 50% of patients required rescue medication after 60 minutes for both treatments (R2 = .498, p = .001), and 66% required rescue medication based on pain level and physiological measures (R2 = .662, p < .008). Based on these results, cryotherapy is an alternative treatment that may be helpful to some patients with ECT-induced headaches. PMID- 15884477 TI - Mid-life widows' narratives of support and non-support. AB - 1. People who simply listened and did not try to give unsolicited advice or control the conversation were perceived by mid-life widows as "supportive listeners." 2. Nurses are in pivotal roles to educate and influence others about the grieving process. 3. Nurses can serve as advocates for widows in using, and teaching others to use, therapeutic communication. 4. Each widow's experience and grieving trajectory is unique. PMID- 15884478 TI - Summary health statistics for the U.S. population: National Health Interview Survey, 2003. AB - OBJECTIVES: This report presents both age-adjusted and unadjusted health statistics from the 2003 National Health Interview Survey (NHIS) for the civilian noninstitutionalized population of the United States, classified by age, sex, race and Hispanic or Latino origin, family income, poverty status, education, place of residence, region of residence, and where appropriate, health insurance coverage. The topics covered are health status and limitations in activities, special education or early intervention services, injuries and poisonings, health care access and utilization, and health insurance coverage. SOURCE OF DATA: The NHIS is a household, multistage probability sample survey conducted annually by interviewers of the U.S. Census Bureau for the Centers for Disease Control and Prevention's National Center for Health Statistics. In 2003, household interviews were completed for 92,148 persons living in 35,921 households, reflecting a household response rate of 89.2%. SELECTED HIGHLIGHTS: Nearly 7 in 10 persons were in excellent or very good health in 2003. About 34 million persons (12%) were limited in their usual activities due to one or more chronic health conditions, and about 4 million persons (2%) required the help of another person with activities of daily living. About 6% of children received special education or early intervention services. Among persons under age 65 years, about 41 million (17%) did not have any health insurance coverage. The most common reason for lacking health insurance was cost, followed by a change in employment. PMID- 15884479 TI - We built it and they did not come: knowledge and attitudes of baccalaureate nursing students toward the elderly. AB - A pilot program in gerontological nursing was developed for senior baccalaureate nursing students but could not be implemented due to lack of student interest. As a consequence, the authors conducted a descriptive survey research study to determine the knowledge base and attitudes of junior and senior baccalaureate nursing students toward older adults. A sample of 55 students was surveyed using two instruments: Palmore's revised Facts on Aging Quiz (FAQ 1) and Kogan's Attitudes Toward Old People Scale (KOP). While neither the junior or senior student group scored high on the FAQ 1, an analysis of variance revealed a significant difference between the groups in overall knowledge about the elderly. The mean KOP score of neither group indicated a high positive attitude toward the elderly and there was no significant difference between the groups in this area. The relationships between participants' demographic characteristics and KOP results also were explored. Findings support research indicating that nursing students often lack knowledge of the elderly and need opportunities to develop positive attitudes toward them. Implications are identified that relate to curriculum development, students, and faculty. PMID- 15884480 TI - Gardening: a strategy for health promotion in older women. AB - One of the goals of Healthy People 2010 is improved cognitive status of older adults. Preliminary research has identified gardening as an activity that may be cognitively protective. Clarification of gardening as a concept is a first step toward the development of theory that will enable nurses to develop interventions related to gardening. The purpose of this study was to describe the phenomenon of gardening. Using a phenomenological methodology, interviews with five older women were analyzed using Colaizzi's approach. Four themes emerged: "Gardening is challenge and work," "Gardening is connection," "Gardening is continuous learning," and "Gardening is sensory and aesthetic experience." The phenomenon of gardening is analogous to the relationship between a spider and its web, linking internal and external environments and providing support over a lifetime. It appears that the gardening experience, as an evolving lifelong process, sustains older women in their cognitive and spiritual development. PMID- 15884481 TI - Revolutionizing prenatal care: new evidence-based prenatal care delivery models. AB - Much of prenatal care is based on tradition and expert opinion rather than on sound scientific evidence. With the increased emphasis on providing evidence based prenatal care, new research-based models are emerging. This article describes two new models of prenatal care delivery and the evidence supporting them. A model of reduced-frequency prenatal visits is adapted from the U.S. Department of Health and Human Services (HHS) Expert Panel on Prenatal Care (1989) recommendations that healthy, pregnant women who are at low risk for pregnancy complications may attend fewer visits without negative consequences. Another emerging model of group prenatal care, CenteringPregnancy, integrates group support with prenatal care. It is important that healthcare providers are aware of these models in order to offer the highest quality, evidence-based care to pregnant women. PMID- 15884482 TI - The influence of individualized music on patients in physical restraints: a pilot study. AB - This pilot study explored the relationship between listening to preferred music and the behavioral responses of patients who are physically restrained. Thirty patients, ranging in age from 65 to 93, participated in one of three groups. The first group included patients who were out of restraining devices while listening to preferred music. Patients in the second group were out of restraining devices and not exposed to music. The third group comprised patients who were in restraining devices while listening to preferred music. Listening to preferred music had no significant effect on decreasing patients' negative behaviors or on increasing positive behaviors observed during the intervention phase of the study. The higher mean scores for positive behaviors and lower mean scores for negative behaviors for the first group may indicate some benefits to patients who are out of restraints and listening to preferred music. PMID- 15884483 TI - Strategies for advancing evidence-based practice in clinical settings. AB - Evidence-based practice (EBP) is a problem-solving approach that incorporates the best available scientific evidence, clinicians' expertise, and patients' preferences and values. Melnyk and Fineout-Overholt have developed the ARCC (Advancing Research and Clinical practice through close Collaboration) model for the purpose of implementing EBP. A pilot study was conducted to test the ARCC model at two acute-care sites. This article shares information learned from the pilot study about what is necessary for successful implementation of EBP in the acute-care setting. These essentials include identifying EBP champions, redefining nurses' roles to include EBP activities, allocating time and money to the EBP process, and creating an organizational culture that fosters EBP. In addition, practical strategies for implementing EBP are presented to encourage implementation of EBP. PMID- 15884484 TI - [Chronic obstructive pulmonary disease]. PMID- 15884485 TI - [Epidemiology and prevention of COPD]. AB - The prevalence of COPD when using spirometry data lies between 4% and 10%. However, health statistics do not fully account for COPD patients either, given the underdiagnosis of this disease reported between 30% and 50%. In the late stage, COPD is associated with a great impact on quality of life and repeated costly hospital stays. Given that COPD in its early stage would be easily preventable by means of smoking cessation, measures of early detection are urgently needed to prevent the COPD epidemic of the years to come. PMID- 15884486 TI - [Chronic obstructive pulmonary disease (COPD): expression of a chronic inflammatory reaction]. AB - Our understanding of chronic inflammation in COPD has been profoundly changed as a result of findings which suggest that airway inflammation in COPD consists not only of a chronic cellular inflammatory reaction dominated by lymphocytes and neutrophil granulocytes, but also of an intensified secondary wound healing reaction. Unfortunately, little is known about the impact of both reactions on the maintenance of chronic airway inflammation in COPD. Systematic biological evaluation under clinical circumstances will be necessary in order to realize new therapeutic options in COPD. PMID- 15884487 TI - [The pathobiology of COPD]. AB - Chronic obstructive pulmonary disease (COPD) is characterised by a chronic inflammation in the pulmonary tissue. The disease is associated with a switch from a self-limiting inflammatory response, mainly initiated by smoke inhalation, to a chronic persistent inflammatory response after prolonged interaction with cigarette smoke. The development and progression of chronic obstructive pulmonary disease have been associated with increased oxidative stress and reduced antioxidant resources. As a result, the antioxidant capacity decreases in COPD patients. Additionally, there is an imbalance between the release of proteases and of endogenous anti-protease enzymes that prevent elastin digestion. The inflammatory basis of COPD is now well established. In patients with COPD increased numbers of macrophages and neutrophils have been found in induced sputum, the former predominating in mild disease. Biopsy data confirm these observations but also demonstrate increased numbers of CD8-positive T- lymphocytes in the airway wall, which may have a role in the regulation of the inflammatory response to cigarette smoke. The extent of the inflammatory reaction is correlated with the severity of the disease. An imbalance between pro- and anti-inflammatory cytokines may favour this process. Several mediators are involved in COPD including leukotriene B4 (LTB4), tumor necrosis factor alpha (TNF-alpha), and interleukin8 (IL-8). They are selective attractants of neutrophils and regulate the ongoing inflammatory process. The whole process would be predicted to be self-perpetuating leading to a chronic inflammatory state with associated airway remodelling and progressive lung function decline. PMID- 15884488 TI - [The diagnosis of COPD]. AB - Lung function testing (spirometry) is crucial for the diagnosis of COPD, as irreversible airway obstruction is the main feature of chronic airway inflammation. Spirometry not only is essential in making the diagnosis but also in grading the disease according to FEV1 measurements. Therapeutic interventions and prognostic evaluations are made according to the grading of the disease. Diagnostic procedures aim to evaluate symptoms and disability in the course of COPD. The most important influence on the course of the disease are acute exacerbations. The diagnosis of acute exacerbations is based on clinical observations of sputum production, cough and dyspnoea. Grading of exacerbations according to the severity of symptoms is important for the treatment and in particular for the need of hospitalisation. Exclusion of other lung diseases with similar symptoms necessitates a number of other examinations. Optimal treatment of COPD needs clinical and objective documentation of the course of the disease. Not only spirometry but also quantification of clinical symptoms and exercise capacity measurements are appropriate approaches to follow this chronic disease. PMID- 15884489 TI - [Pharmacological management of stable COPD]. AB - Pharmacotherapy for stable COPD depends on the severity of the disease. Thus, decisions on therapeutic interventions should be based not solely on the degree of airflow obstruction but also on symptoms, exercise tolerance, frequency and severity of exacerbations, and health-related quality of life. The progressive nature of COPD necessitates a stepwise increase of medication (step-up approach) depending on the severity of the disease. Although none of the currently available medications has been shown to modify the abnormal reaction to inhaled noxious particles and gases that lead to COPD, pharmacotherapy has beneficial effects on the clinical course of the disease by controlling symptoms, increasing exercise tolerance and improving general health status. PMID- 15884490 TI - [COPD-rehabilitation]. AB - Rehabilitation of COPD-patients is an important part of the therapeutic management. The effects of endurance- and resistance-training as well as respiratory muscle-training are evident. Smoking cessation therapy is standardized, effective and cheap, and is the prerequisite of a successful COPD management. Patients with severe or very severe COPD not only have to inhale their medication exactly, but also have to undertake physical exercise and optimize their body weight. This cannot normally be achieved without adequate education. A structured patients' education optimizes all therapeutic action and is an integral part of the management of COPD. PMID- 15884491 TI - [Medical therapy by training in the rehabilitation of patients with COPD]. AB - Medical therapy by training means physical training for patients as part of a medical therapy based on a physician's recommendation, with clear indications and with clear therapeutic aims. It has to be clearly differentiated from sport and is not an alternative medicine. There is a lot of evidence that physical training can improve endurance and strength of patients with COPD. Especially in severe COPD, the VO2max, as the parameter for the endurance capacity, is the best single predictor for survival, better than every other value of lungfunction. The rules of training concerning endurance and strength in patients with COPD are presented. PMID- 15884492 TI - [Ventilation therapy for patients with COPD]. AB - Acute ventilatory failure in COPD is caused by impaired respiratory mechanics and an imbalance of capacity and load of the respiratory muscles. Continuous positive airway pressure (CPAP), pressure support ventilation (PSV), proportional assist ventilation (PAV) or controlled mechanical ventilation (CMV) are effective in unloading the respiratory pump. CPAP reduces the inspiratory pressure time product by reducing the elastic work of breathing due to intrinsic PEEP (PEEPi). PSV and PAV reduce the work of breathing and additionally improve alveolar ventilation and CO2 elimination. In spontaneous breathing patients these modes of ventilation should be applied non invasively by using a face mask. In CMV, hyperinflation should be avoided by choice of low tidal volumes and long exspiratory times (low respiratory rate, I:E ratio 1:3, 1:4). In the weaning from CMV, PSV or PAV and CPAP is used. There seems to be a beneficial effect in nocturnal intermittent nonivasive ventilation (NPPV) in chronic hypercapnic stable COPD patients with documented hypoventilation during the night, which is reversible by NPPV. PMID- 15884493 TI - [Is imaging possible for otosclerosis? Characteristics of the disease and a survey of the clinical relevance of imaging]. AB - Otosclerosis is a localized progressive disease of bone remodeling within the otic capsule of the human temporal bone. Histomorphologically, active cochlea otosclerosis (otospongiosis) is diagnosed in the presence of foci of demineralization in the otic capsule. The clinical symptoms of otosclerosis are associated with mixed and sensorineural hearing loss. With recent technological advances, diagnostic imaging of the inner ear is becoming more and more important in the evaluation of diseases affecting the cochlea. Studies could demonstrate that otosclerosis has a characteristic appearance on high-resolution computed tomography (HRCT), producing a distinctive pericochlear hypodense double ring. Its appearance on MRI is not as readily appreciated, producing a ring of intermediate signals in the pericochlear and perilabyrinthine regions on T1 weighted images, demonstrating mild to moderate enhancement after gadolinium administration. The ethiology and pathophysiology of otosclerosis are described and a review of the literature is given to illustrate the clinical relevance of imaging in otosclerosis. PMID- 15884494 TI - [Constipation and fecal incontinence: when there is nothing--or too much passes]. PMID- 15884495 TI - [Therapeutic possibilities for incontinence. Biofeedback strengthens the sphincter]. PMID- 15884496 TI - [Alternatives to the conventional hearing aid: natural hearing quality with partial implant? (interview by Dr. Judith Neumaier)]. PMID- 15884497 TI - [1st heart catheter, 1st coronary dilatation, 1st resorbable stent. Renewed breakthroughs in German research]. PMID- 15884498 TI - [Psyche and soma between fitness and fatigue. Is the drive for youth making us ill?]. PMID- 15884499 TI - [Pain patients in the nursing care setting. Relief of pain is a human right]. PMID- 15884500 TI - [Diagnosis and treatment of pain in the elderly patient]. AB - For the diagnosis of pain in the elderly patient, special instruments are available for measuring cognitive and sensory deficits. With regard to treatment, too, age-specific peculiarities must be taken into account. Musculoskeletal diseases, and especially osteoarthritic conditions, are the most common pathologies that are the cause of pain in old age. To a particular degree they result in a restriction of mobility and pose a threat to the patient's independence. Physical inactivity is a major risk factor for the development of chronicity and for patient withdrawal. An improvement in the patient's training status can be achieved by pharmacological and physical treatment applied in association with active physical exercise and psychological measures. PMID- 15884501 TI - [Pain management in nursing care--experts standard]. AB - The experts standard of pain management in nursing care regulates the measurement and recording of pain, medicational and non-medicational measures for the treatment and prevention of pain, the treatment of potential side effects of analgesics, and the counseling and training of nursing personnel. As such, it impacts on earlier shortcomings of interprofessional pain management, provides for therapeutic certainty, and improves the situation of those requiring permanent nursing care. PMID- 15884502 TI - [The problem of pain in a German nursing home]. AB - The experience of, and reaction to, pain by the inhabitants of a nursing home (n = 148) were evaluated. They were invited to score their pain status with the aid of a verbal scale, and/or the nursing staff were asked to estimate it on the basis of a numerical rating scale. The regular and as-required prescription of painkillers was also recorded. Acute attacks of pain were symptoms of sometimes life-threatening diseases, and were scored at the highest level of severity. Patients suffering from chronic pain were most often treated with opioid analgesics. Recurrent attacks of pain were preferentially treated with NSAIDs or metamizol. Pain experienced only during nursing measures or while taking exercise was treated too infrequently. When appropriately trained, the nursing staff are well prepared to establish and document the painful situation, and to rapidly identify new pain syndromes. Regular evaluation should be a standard practice of nursing care. PMID- 15884503 TI - [Threshold and target blood pressures for treating hypertension. The facts]. PMID- 15884504 TI - [Whipple's disease: typical cases and review]. PMID- 15884505 TI - [What is not in the manual so far: emergency on call duty calculated according to new the new EBM]. PMID- 15884506 TI - [The ordination complex in emergency on call duty]. PMID- 15884507 TI - [A few colleagues already use the modern method. Patients schedule office appointments on the internet]. PMID- 15884508 TI - [Diagnosis at a glance. After oral sex an ulcer on the tongue]. PMID- 15884509 TI - Will the cytokine be the future therapy of uveitis? PMID- 15884510 TI - Glaucoma therapy: "above all do no harm". PMID- 15884511 TI - Role of cytokines in experimental and clinical uveitis. AB - Uveitis is a major cause of visual impairment. Experimental autoimmune uveitis mimics the clinical conditions of posterior uveitis in many ways. T-cells (particularly CD4+ helper-T-cells) have been shown to play an important role in pathogenesis of experimental and clinical uveitis. Based on the pattern of cytokine they secrete, CD4+ helper cells have been divided into Th1 and Th2 subsets. Various Th1 and Th2 cytokines appear to be involved in the pathogenesis and/or recovery from uveitis. This article discusses in detail the uveitopathogenic and therapeutic potential of Th1 and Th2 cytokines in experimental and clinical uveitis. PMID- 15884512 TI - Assessing the role of subconjunctival versus intrascleral application of mitomycin C in high-risk trabeculectomies. AB - PURPOSE: To compare the efficacy and safety of subconjunctival and intrascleral applications of mitomycin C (MMC) in trabeculectomy for high-risk glaucomas. METHODOLOGY: A randomized prospective clinical study was conducted on 41 consecutive eyes with a high risk of glaucoma surgery failure. Patients were randomized to trabeculectomy and application of subconjunctival MMC or to trabeculectomy and application of intrascleral MMC. MMC solution 0.2 mg/ml was applied for 3 minutes under the conjunctival flap overlying the proposed site of trabeculectomy in Group I (n=21), or intrasclerally under the superficial scleral flap in Group II (n=20) RESULTS: After a follow-up of one year, the intraocular pressure (IOP) decreased from a mean basal IOP of 33.0 +/- 8.4 mm Hg to 12.56 +/- 2.54 mm Hg in Group I and from 30.9 +/- 6.6 mm Hg to 11.6 +/- 2.21 mm Hg in Group II. The IOP was 6-21 mmHg, without medication, in 90.5 % of the eyes in Group I and 75 % of the eyes Group II. Ocular hypotony, hypotony maculopathy, choroidal detachment and a shallow anterior chamber were more frequent with the intrascleral application of MMC during trabeculectomy, but the difference was not statistically significant. The overall success of the surgery at one year, i.e., achieving an IOP of 6-21 mmHg and a stable vision, (reduction in visual acuity of < or = 2 lines), was 90.5% in Group I and 75 % in Group II. CONCLUSION: No significant difference was seen in overall success or complication between subconjunctival and intrascleral application of MMC-augmented trabeculectomies in glaucomatous eyes at high risk of surgical failure. PMID- 15884513 TI - Threshold retinopathy of prematurity: ocular changes and sequelae following cryotherapy. AB - PURPOSE: To determine ocular changes and sequelae following cryotherapy for threshold retinopathy of prematurity (ROP). METHODS: This is a retrospective study of 49 eyes of 26 premature babies with threshold ROP treated with cryotherapy between 1995 and 1998. All eyes included in the study had favourable structural outcome after cryotherapy. Follow-up examination of all babies was done 12 - 62 months (average 28 months) after cryotherapy. Visual axis, fixation pattern, anterior segment examination, cycloplegic refraction and dilated fundus examination with indirect ophthalmoscopy were undertaken in all eyes during follow-up. RESULTS: Posterior pole retinal residuae observed following cryotherapy were tortousity of blood vessels in 32 (65.3%), narrow temporal arcade in 22 (44.89%), temporal crescent in 17 (34.69%), disc drag in 13 (26.53%) and macular heterotopia in 7 (14.28%) eyes. Myopia was observed in 20 (40.82%) eyes and strabismus in 5 (19.23%) babies. The significant risk factor for ocular changes was ROP with more clock hours of involvement (p < 0.05). Higher period of gestation was associated with posterior pole changes (p< 0.05). CONCLUSIONS: All premature babies with threshold ROP treated with cryotherapy require frequent and long-term follow up to look for retinal residuae, refractive status, and ocular motility disorders. PMID- 15884514 TI - A prospective study of 413 cases of lens-induced glaucoma in Nepal. AB - PURPOSE: To determine the frequency and types of lens-induced glaucoma (LIG), reasons for late presentation and outcome of current management. METHODS: Prospective case series of 413 patients/eyes with LIG over a 12-month period in 1998; 311 of these patients underwent cataract surgery. Visual acuity and intraocular pressure (IOP) were pre- and postoperatively assessed. RESULTS: Four hundred and thirteen (1.5%) of 27,073 senile cataracts seen in the outpatient department of Sagarmatha Choudhary Eye Hospital, Lahan, Nepal presented with LIG. There were 298 (72%) phacomorphic cases and 115 (28%) phacolytic glaucoma. Pain for more than 10 days was reported by 293 (71%) patients. The majority, 258 (62.4%), travelled a distance of more than 100 kms to the hospital. The major reasons for late presentation were "no escort" in 143 (34.6%) and "lack of money" in 128 (31.0%) cases. At presentation the IOP was more than 30 mm Hg in 327 (79%) eyes. Following cataract surgery, 251 (80.7%) had 21 mm Hg or less at discharge. The visual acuity was hand-movement or less before surgery in all eyes; at discharge 120 of 311 operated eyes (38.6%) achieved 6/60 or better, 97 (31.2%) less than 6/60, and 94 (30.2%) less than 3/60. The main causes for poor outcome in 94 cases were optic atrophy in 32 (34%) eyes, uveitis in 25 (26.6%)eyes and corneal oedema in 24 (25.5%) eyes. CONCLUSION: The results highlight the importance of early diagnosis and treatment of visually disabling cataract. There is a need to educate both the patient and the cataract surgeon of the dangers of lens-induced glaucoma and of the poor outcome if treatment is delayed. PMID- 15884515 TI - Scleral suspension pars-plana lensectomy for ectopia lentis followed by suture fixation of intraocular lens. AB - PURPOSE: To describe a simple technique of scleral suspension-pars plana lensectomy (SS-PPL) in acquired and congenital ectopia lentis and scleral fixation of intraocular lens (IOL). MATERIALS AND METHODS: Twenty eyes of 16 patients (12 unilateral and 4 bilateral cases of "essential familial lens subluxation") aged 10-40 years (mean 25 years) underwent SS-PPL with implantation of scleral fixated IOL. Indications for surgery were best-corrected visual acuity < 6/18, bisection of pupil by the lens, and lens-induced glaucoma. Prerequisites for SS-PPL were, visibility of part of the lens in the pupillary area and soft lens. RESULTS: Postoperative visual acuity ranged from 6/6 - 6/36. Lens tilt in 3 cases(15%) and small decentration in 2 cases(10%) were seen; however these did not seriously compromise the visual result. Scant vitreous bleeding on the first postoperative day was seen in 3 cases (15%). CONCLUSION: The advantages of the scleral suspension of subluxated lens prior to lensectomy include stabilization; it allows proper viewing of the lens, avoids injury to the iris and ciliary body during lensectomy and reduces the possibility of dislocation of the lens. PMID- 15884516 TI - Fish hook injury to the eyelid - an unusual case. AB - Fishing is recreational activity enjoyed by millions of people worldwide. Although uncommon in developing countries it is a popular pastime in the Western world. and, given this popularity, a number of associated personal injuries are probably inevitable. We report a case of fish hook injury to the eyelid and discuss the various techniques available for removal of fish hooks. The terminologies associated with various parts of the fish hook are also illustrated. PMID- 15884517 TI - Conjunctival amyloidosis of both eyelids. AB - Amyloidosis of the palpebral conjunctiva is a rare condition which results in chronic ocular discomfort. We present our experience with this rare patient in whom both upper and lower palpebral conjunctivae were involved, but no systemic amyloidosis. PMID- 15884518 TI - Multiple cranial nerve palsy in an HIV-positive patient. AB - We report the case of a 32 -year- old HIV-positive Indian male who presented with sixth nerve (bilateral), ninth, tenth and twelfth nerve palsies; cerebellar and posterior column involvement. CT scan showed gyriform enhancement in the right occipital lobe and nodular leptomeningeal enhancement in the left frontal lobe. Cytomegalovirus serology was positive and the patient was treated as presumed CMV. HIV can present with multiple cranial neuropathy and varied neurological involvement. PMID- 15884519 TI - Childhood proptosis: the invaluable but overlooked peripheral blood smear. AB - Two cases of granulocytic sarcoma presenting as childhood proptosis are described. This highlights the significance of peripheral blood smear examination in establishing the diagnosis of Acute Myeloid Leukaemia (AML). PMID- 15884520 TI - Interpreting automated perimetry. AB - Visual field testing is mandatory for many ophthalmic conditions including glaucoma. The current gold standard for visual field testing is automated perimetry. In this article we familiarize the reader with the components of an automated perimetry printout. We describe a systematic approach that leads to a thorough interpretation of the printout. With the help of examples the reader should be able to learn to identify a normal field, detect the presence of a field defect, determine whether it is due to glaucoma, and establish progression, if any. PMID- 15884521 TI - New approaches in the management of choroidal neovascular membrane in age-related macular degeneration. PMID- 15884522 TI - [Dynamic loads at knee joint of trans-tibial amputee on different terrains]. AB - Dynamic loads at knee joint of amputee are fundamental for rehabilitation of knee injury and prosthesis design. In this paper, a 3-D model for calculation of dynamic load at knee joint of trans-tibial amputee was developed. Gait analysis was done on three terrains including normal level walking, upstairs and downstairs. Dynamic loads at knee joint were calculated during one gait cycle. The results show that gait patterns and dynamic loads at knee joint were different among these three terrains. Although the general waveforms were about the same, the motion range of knee joint, ground reaction forces and loads at knee joint when walking upstairs or downstairs were larger than those in a normal level walking. The quantitative findings provide the theoretical basis of gait analysis and prosthesis design for trans-tibial amputee. PMID- 15884523 TI - [Effects of wall shear stress on the morphology and permeability of endothelial cells in stenotic rabbit abdominal aorta]. AB - Stenosis with 55.2% cross section area reduction was introduced into the rabbit aorta. Using Evans blue dye and scanning microscope, we observed the morphology of endothelial cells and the permeability of endothelium to albumin in the stenotic aorta. Numerical simulation of blood flow in the stenotic aorta was performed to obtain the distribution of wall shear rate. The results showed that in the immediately proximal and distal vicinity of stenosis, blood flow was disturbed significantly, resulting in apparent changes in the morphology of endothelial cells and the permeability of endothelium to albumin. These changes were not only attributed to the value of wall shear stress, but also attributed to the flow pattern in the stenosis. The result therefore is in good consistent with the clinical observation that atherosclerosis often occurs in the areas where blood flow is disturbed and flow separation occurs. PMID- 15884524 TI - [Effect of ERK1/2 on low shear stress-induced expression of IL-8 mRNA in human endothelial cells]. AB - Fluid shear stress plays an important role in many physiological and pathophysiological processes of the cardiovascular system. It modulates vascular function and structure via stimulating mechanosensitive endothelial cell signal events. Previous studies have identified that the exposure of vascular endothelial cells to fluid mechanical forces can modulate the expressions of many genes, including IL-8 gene. In order to gain an insight into the role of extracellular signal regulated kinase (ERK1/2) signal pathway in the expression of IL-8 mRNA in human umbilical vein endothelial cells (HUVECs) under the stimulation by low shear stress (4.20 dyne/cm2), we employed Western blot to measure phosphorylation of ERK1/2 and used quantitative reversal transcription polymerase chain reaction (qRT-PCR) to assay the expression of IL-8 mRNA. The results showed: (1) Shear stress could activate ERK1/2 with a rapid, biphasic time course (maximum by 10 min and basal by 2 h); the treatment of HUVECs with Genistein (a highly specific inhibitor of tyrosine protein kinase, TPK) or PD98059 (the inhibitor of mitogen-activated protein/extracellular signal regulated kinase kinase, MEK) culd prevent shear-dependent activation of ERK1/2; (2) When treated with Genistein or PD98059, significant inhibition of IL-8 mRNA expression induced by low shear stress was observed in HUVECs. This in vitro study demonstrates that ERK1/2 plays an important role in IL-8 mRNA expression induced by low shear stress. PMID- 15884525 TI - [Energy mechanism of effects of carbon phase components on hemocompatibility for DLC]. AB - The correlations between surface energy parameters and carbon phase components in six diamond like carbon film (DLC) samples made in different ways and processing conditions were further investigated using the analysis of T-type correlation degree in the Grey system theory based on our earlier studies such as the determination of carbon phase and surface energy parameters, the analyses of carbon phase components, surface energy parameters and adhesive characteristic of platelets for DLC. The results showed: (1) as a whole critical surface tension has the closest relation with carbon phase components, chromatic dispersion branch of surface tension takes the second place, but for the other four parameters, the correlation is weak; (2) DLC phase has larger negative correlation (degrees -0.57, -0.33) with critical surface tension and chromatic dispersion branch of surface tension, while its correlation degrees with the other four parameters related to polarity are all positive values smaller than 0.20; (3) C-H phase and C-O phase have larger positive correlation (degrees 0.48, 0.25) with critical surface tension. We have come to three conclusions (1) DLC phase plays a dominant part in hemocompatibilioty of DLC by powerfully decreasing humidification and limitedly increasing polarity; (2) the energy mechanism of platelet deformation enhanced by C-H phase and C-O phase involves increasing the critical surface tension of DLC; (3) the hemocompatibility of DLC can be evaluated by using the critical surface tension as index and using the content of DLC phase and additional content limitation of C-H phase and C-O phase as standard. This study has provided a theoretical basis for evaluating the hemocompatibility of DLC based on surface properties. PMID- 15884526 TI - [Preparation and properties of hydroxyapatite/epoxy composite]. AB - The hydroxyapatite (HA) powder surface modified with silane coupling agent was used to prepare HA/epoxy composite. It was found that silane has greatly improved the dispersion of HA in epoxy. The composite has good in vitro bioactivity and biocompatibility with 40 wt% HA, and the flexural modulus is close to that of natural bone, but its strength is lower than that of natural bone. So the composite needs further reinforcement in some way or other. PMID- 15884527 TI - [Research of osteoblast-induced rat mesenchymal stem cells cocultured with beta TCP/PLLA composite of different ratio]. AB - There are three key factors in tissue engineering: seeding cells, scaffold and their interaction. Although mesenchymal stem cells (MSCs) are potential seeding cells, the problem of what phase MSCs should be used is not yet solved. On the other hand, degradable porous scaffolds which have good mechanics and good biocompatibility are preferred. To choose the optimum seeding cells and the suitable ratio of beta-TCP/PLLA porous scaffold, we observed the phenotype of the male SD rat's osteoblastic MSCs and detected the amount of alkaline phosphatase, osteocalcin and type I collagen secreted by the osteoblastic rMSCs in different phase. About 10, 14 and 20 days after induction, the induced cells came into proliferative phase, matrix synthesis phase and mineralization phase, respectively. Then we chose the suitable cells and seeded them on beta-TCP/PLLA composite scaffolds with different ratios (beta-TCP/PLLA = 1:1; beta-TCP/PLLA = 1:2; and beta-TCP/PLLA = 2:1). Fluorescence microscope, scanning electron microscope and MTT assay were used to observe and to detect the biocompatibility of the scaffolds. The results indicated that all of these materials have biocompatibility to some extent. Cells can grow well on all of the scaffolds. However, scaffold beta-TCP/PLLA = 2:1 seems to be a more suitable tissue engineering scaffold on account of its minimal influence on cell growth and differentiation. PMID- 15884528 TI - [Hydroxyapatite bioactive coating on carbon/carbon composites]. AB - A simple plasma spraying method was employed in coating hydroxyapaptite (HA) on to carbon/carbon composites (C/C composites). The morphology of the coating was examined under scanning electron microscope (SEM). The phase constitutions of the HA coating were determined by X-ray diffractometer (XRD). The shear strength of the HA coating-C/C composite substrates was detected. A hydroxyapatite coating with rough surface was observed. A considerable amount of amorphous phase appeared as a result from the coating process, which could be transformed into the morphous phase crystalline HA after subsequent heat treatment. The shear strength between the HA coating and C/C composite substrates was 7.15 MPa. PMID- 15884529 TI - [Expression and immunity reaction of a novel gene OmpL17 of the strong virulent L. interrogans serovar Lai in China]. AB - This study was conducted to potentiate the expression of outer membrane protein OmpL17 of the strong virulent L. interrogans serovar Lai and investigate its immunogenicity in rabbits. The OmpL17 was cloned into prokaryotic expression vector pGEX-1lambdaT. The recombination expression plasmid pGEX-OmpL17 was transformed into E. Coli JM109. The GST fused protein GST-OmpL17 was expressed after induction by IPTG, then GST-tag was by thrombin and purified using Bulk GST purification Modules. SDS-PAGE and Western blotting analysis indicated that the molecular weight of GST-OmpL17 and OmpL17 was about 54 KDa and 28 KDa respectively. The outer membrane protein OmpL17 was subcutaneously injected into rabbits and high titre anti-OmpL17 antibody was obtained (1:4896) which could conjugate specifical with OmpL17. In conclusion, OmpL17 and specifical anti OmpL17 antibody were obtained, which provided an experimental basis for researching pathogenic effect and immunity functions of OmpL17. PMID- 15884530 TI - [A study on the repair of bone defects with deproteinized bone surrounded by titanium mesh and osteoblasts]. AB - To investigate the ability of composite graft of osteobalsts and deproteinized bone-titanium mesh (DPB-TM) scaffold to repair cranial bone defect. 30 rabbits were randomly divided into 3 groups. The passage 3 fetal rabbit osteoblasts were seeded into porous DPB-TM scaffolds at the density of 5 x 10(6) ml(-1) as the experimental group. The same defects were respectively reconstructed by DPB-TM or osteoblasts as the control groups. After 12 weeks, the result was evaluated by three-dimensional computed tomographic scanning, gross inspection, scanning electron microscopy, histological examination and mechanics test, respectively. In the experimental group, bone trabecula was observed to pass the defect and interface was mixed. No demarcation between the region of the bone defect and the normal bone was observed. There was plenty of new bone on the scaffold. Part of the scaffold was absorbed. In view of mechanics, the intensity of artificial bone (18.93+/-1.12 MPa) was higher than that of normal bone (16.96+/-1.60 MPa) (P<0.05). In the control groups, only fibrous tissue was observed in the defect region, there was no new bone formation. The tissue engineering bone constructed by osteoblasts and DPB-TM scaffold can be applied to the repair of bone defect. PMID- 15884531 TI - [Interleukin-8 protein and gene expression in atherosclerotic lesions of hyperlipemia rabbits]. AB - Interleukin-8 is CXC chemokine that is initially discovered using chemotaxis and the activation of neutrophils and induces the migration and proliferation of smooth muscle cells. Interleukin-8 is a potent angiogenic factor that may play a role in atherosclerosis. To establish the temporal correlation between IL-8 expression and plaque development, we examined the expression during atherosclerosis of hyperlipemia rabbits using immunohistochemistry, ELISA, in situ hybridization. By location of immunohistochemistry, the expression of IL-8 protein increased obviously in intima of hyperlipemia rabbits at 8 and 12 week. Quantitative analysis of the expression of IL-8 Immunohistochemistry indicated that positive area of AS model was 4.48 times and 8.76 times that of control group at 8 and 12 week. The valuation of IOD of AS model was 4.16 times and 4.36 times that of control group at 8 and 12 week. By specific ELISA, the ratio of the IL-8 protein to total protein of AS model was 1.84 times and 2.06 times that of control group at 8 and 12 week. By location of in situ hybridization, positive location was strong in intima of hyperlipemia rabbits at 8 week. We observed the dynamic alteration of interleukin-8 protein and gene expression in atherosclerotic lesions of hyperlipemia rabbits with establishing model. Interleukin-8 protein and gene expression was up-regulation in the development of fatty streaks in hyperlipemia rabbit. PMID- 15884532 TI - [Neonatal murine keratinocytes split express CD80/CD86 upon culture]. AB - It was previously thought that keratinocytes did not express the CD80 and CD86 which provide the most important costimulatory signals for the antigen-specific T cell activation. The cultured keratinocytes allografts were initially accepted, but eventually, all grafted donor cells were gradually replaced by recipient cells. The precise mechanisms are not very clear. In this study, neonatal murine keratinocytes were cultured for 7 days, the results of flow cytometry and confocal microscopy showed that CD80 could be detected on keratinocytes, while CD86 could not be detected all the time. RT-PCR analysis confirmed this result. The expression level of the CD80 mRNA amplified significantly from day 1 to day 7, as expression of the control beta-actin, but CD86 was not detected. Mixed Lymphocyte Reaction (MLR) showed that keratinocytes cultured with 10% serum for 7 d stimulated effectively allogeneic rather than syngeneic T cell proliferation. This study demonstrated for the first time that costimulatory molecule CD80 can be expressed on keratinocytes in vitro. These data provided an alternative explanation for the ultimate rejection of allogeneic keratinocytes in which keratinocytes act as antigen-presenting cells. PMID- 15884533 TI - [Overexpression of TGFbeta1 increases ECM expression and adhesion of endothelial cells on matrix]. AB - This study was aimed to examine the effectiveness of a gene transfer of human TGFbeta1 gene into endothelial cells and to determine whether TGFbeta1 increases ECM expression of endothelial cells. With the help of DOTAP, endothelial cells were transfected with pMAMneoTGFbeta1. The positive cell clones were selected with G418. The stable transfection and expression of TGFbeta1 in the endothelial cells were determined by immunofluorescence analysis. The expression levels of collagen I and fibronectin in the transfected and untransfected endothelial cells were determined by Western blot. The adhesion force between endothelial cells and matrix was determined by a micropipette technical system. The results showed abundant TGFbeta1 stable expression in the endothelial cells. TGFbeta1 gene was noted to increase collagen I and fibronectin expression and increase the adhesion between endothelial cells and matrix. These findings indicated that TGFbeta1 can be used in vascular tissue engineering for the enhancement of endothelial cells adhesion. PMID- 15884534 TI - [Measuring the electricity frequency properties of blood]. AB - In order to understand the electricity frequency specialties of blood, we have developed a wide frequency electricity characteristic testing system and used it to test the amplitude frequency property and phase frequency property of the blood in different states and constituents at 1 Hz to 20 MHz. Further analysis on the results of tests helped us know some important properties of blood at even more microcosmic levels from a new angle. Meanwhile, some problems and considerations on the improvement of the electricity model of biotic tissue and blood were pointed out. (1) From 1 Hz to 5 KHz, the impedance of blood descended 99%. (2) Simple equivalent circuit of resistance and capacitance must be used in series equivalent but not in usual parallel connection equivalent. (3) Experiment indicated, equivalent circuits of blood need more analysis, because simple equivalent circuit of resistance and capacitance is liable to gross error. (4) When the three element model is used for measuring the resistance of inside liquid, capacitance of cell membrane and the resistance of outside liquid of blood, the three testing frequencies must be very similar. PMID- 15884535 TI - [Feasibility of stem cells transplantation through aorta in adriamycin-induced heart failure]. AB - Stem cells transplantation is a promising strategy for treating myocardial infarction and/or chronic heart failure; however, with respect to nonischemic heart failure, there are some limitations inherent in the current methods of transplantation. In this study, we investigated the feasibility of a novel method, i. e. transplantation through the root of aorta when the ascending aorta occluded above the sinus aortae. Japanese white ears rabbits were used as chronic heart failure models by intravenous injection of adriamycin. Autologous bone marrow mononuclear cells (MNC) were infused into the root of aorta when the ascending aorta was occluded by a couple of balloons above the sinus aortae. After 4 weeks, ejection fraction was significantly improved in MNC group. In conclusion, we have developed a unique method for efficient and safe cell transplantation based on infusion in aorta. This method, potentially suitable for nonischemic heart failure and could be used to achieve even and global supply of cells in heart. PMID- 15884536 TI - [Construction and immunological study of recombinant hBD-2/PSMA chimeric protein eukaryotic expressive plasmid]. AB - The recombinant PSMA DNA vaccine for active immunotherapy of prostate cancer was investigated. Two DNA vaccine recombinant plasmids, pcDNA3.1/PSMA and pcDNA3.1/hBD-2-PSMA, were constructed by inserting the hBD-2 gene and PSMA gene into an eukarytoic expression vector pcDNA3.1. Expression of the two recombinants was detected in transfected COS-7 cells and inoculated mouse muscular cells by RT PCR and immunohistochemical method. When immunized with pcDNA3.1/PSMA and pcDNA3.1/hBD-2-PSMA, the immunized BALB/c mice acquired specific antibody and T cell response to PSMA. The quantity of the spleen lymphocytes and their CTL activity against PSMA gene transfected-BALB/3T3 cells significantly increased in the immunized mice, and the CTL activity of lymphocytes from pcDNA3.1/hBD-2-PSMA immunized mice was significantly higher than that of pcDNA3.1/PSMA immunized mice. This result suggests that pcDNA3.1/hBD-2-PSMA would probably be developed as a DNA vaccine for the immunotherapy of prostate cancer. PMID- 15884537 TI - [A biomechanical system that can apply fluid shear stress to osteoclast-like cells in vitro]. AB - On the basis of the well-established principles and techniques about flow chamber, we have designed and made a kind of parallel plate system to apply steady fluid shear stress to osteoclast-like cells etc. in vitro. Biocompatible rubber and metal clamping apparatus made of aluminium alloy are used to seal the flow chamber without changing its even height designed beforehand. The influence of hydrostatic pressure on cells is minimized by adjusting the glass slide and the fluid surface in the upper reservoir to the same horizontal plane. This system can be used to investigate the responses of osteoclast-like cells etc. to fluid shear force in terms of morphology, physiology or biochemistry. PMID- 15884538 TI - [A preoperative planning system for unilateral craniofacial malformation restoration surgery based on the volume measurement of eye sockets]. AB - The surgical treatment of craniofacial malformation is a sophisticated task which needs careful and detailed preoperative planning. However, based on the patient's 2-D CT and X-ray images, the surgeons always feel hard to produce a convincible and satisfying surgical scheme. In this article, a craniofacial surgery simulation system based upon 3-D reconstruction and volume calculation of two eye sockets is presented. By using this system, the surgeon can virtually repose bone fragment according to the D-value between the volumes of two eye sockets. The D value serves as a guideline to direct the bone reposing. The simulated key steps of real surgical procedure are presented, which demonstrate the efficiency of the system. The contrast images between postoperative and preoperative surgery are also given. PMID- 15884539 TI - [Modeling vocal-fold vibration via integrating two-mass model with finite-element method]. AB - Modeling vocal-fold vibration is extremely significant in realizing the vibration properties of human vocal folds and investigating their physiological and pathological characteristics. A combined model presented is two mass-finite element (T-F) model, which integrates all merits of both the finite element method (FEM) model and the asymmetric two-mass model of vocal folds. The high speed glottis graph (HGG) can also be synthesized by the model. The result shows that T-F model can simulate the vibration behavior of normal and pathological vocal folds in a more realistic way with competitively computational speed. Therefore, the T-F model is helpful to gaining a thorough understanding of the vibration properties of vocal folds. PMID- 15884540 TI - [Design and field calculation of coil array for transcranial magnetic stimulation (TMS) based on genetic algorithm]. AB - It is the intent of this paper to locate the activation point in Transcranial Magnetic Stimulation (TMS) efficiently. The schemes of coil array in torus shape is presented to get the electromagnetic field distribution with ideal focusing capability. Then an improved adaptive genetic algorithm (AGA) is applied to the optimization of both value and phase of the current infused in each coil. Based on the calculated results of the optimized current configurations, ideal focusing capability is drawn as contour lines and 3-D mesh charts of magnitude of both magnetic and electric field within the calculation area. It is shown that the coil array has good capability to establish focused shape of electromagnetic distribution. In addition, it is also demonstrated that the coil array has the capability to focus on two or more targets simultaneously. PMID- 15884541 TI - [Spatial reconstruction of human motion utilizing two dimension images and a biomechanical model]. AB - One of the key techniques in the research of human motion analysis is the reconstruction of human spatial motion utilizing the evaluation of the anatomic points positions that can uniquely define the position of all anatomical segments. Upon the bases of direct linear transform and human biomechanical model, the method to reconstruct an image sequence shot by a stationary camera was described. The result of the reconstruction process was a set of data that is kinematically consistent with the biomechanical model used. A new method to calibrate camera was developed which can be completed in any world coordination system according to the practical situation. A practical motion was selected as the investigated subject. The predicting and experimental results were in good agreement. The benefits and shortcoming of the methodology were discussed. PMID- 15884542 TI - [Algorithm for recognizing the markers in human motion detection]. AB - An algorithm for recognizing the markers in human motion detection is proposed in this paper. It includes three parts as follows: (1) an improved template matching method is used to acuminate the peak of the matching result. (2) The weighted average of the cross correlation between target image and template is given in order to make the resolution increase to sub-pixel level. (3) Forecasting the motion of marker and overcoming the hiding of marker are realized basing on the continuity of human motion. The result of experiments shows that the algorithm can recognize the positions of the markers at a relatively higher accuracy and speed. A gait analysis system based on this algorithm has come into use. PMID- 15884543 TI - [Beat to beat analysis of morphological difference of ECG waveforms]. AB - Three different kinds of ECG data including the healthy, angina and myocardial infarction from the ECG database were analyzed to obtain the morphological difference sequence of their P-waves, QRS-wave and T-wave, and their RR interval sequences. Then the variability of these four sequences was analyzed by detrended fluctuation analysis (DFA) method. The results indicate that, the changing pattern in the DFA index of RR interval sequence from the healthy to the morbidity is different from those of the P-wave, QRS-wave and T-wave. The DFA indices of the RR interval sequence are about 1 in all three states, while the DFA indices of the other three sequences are about 0.5 in the healthy and all decrease from the healthy to morbidity. PMID- 15884544 TI - [Fabricating facial prostheses using CAD/CAM and rapid prototyping technique]. AB - At present, the treatments of hemifacial microsomia such as the missing of ear and eye still rely on the skill of technician to make the wax model of contralateral apparatus of patient in China. In this paper, CAD/CAM and rapid prototyping (RP) techniques are integrated to successfully create a silastic prosthesis by using the patient's data of CT or laser scanning. The clinical results suggest that a high accuracy has been achieved in shape, size, and protrusion of the facial prostheses, which indicates that the application of RP techniques in conjunction with CAD/CAM is a suitable approach for fabricating facial prosthesis. PMID- 15884545 TI - [Visualization of regenerated sciatic nerve of rat after injury]. AB - There is close relationship between the morphologic changes of peripheral nerve after injury and its function recovery during regeneration. In our experiment, the sciatic nerve of rats was transected and bridge-connected with silicone tube, and the images of serial slices of different time and different injury parts were taken by micro-photograph system. The volume unit model was applied to rendering the three dimensional (3D) structure of degenerative and regenerative sciatic nerve fiber and its affiliated structure after injury. The 3D images showed that node of Ranvier had not formed and its array was turbulent, both the myelin sheath and the axis-cylinder were thinner, and the collagen fibres had proliferated. PMID- 15884546 TI - [Analysis of millimeter-wave dosimetry in cell culture dishes with finite difference time-domain technique]. AB - Analysis is carried out on millimeter-wave (MMW) dosimetry in culture dishes used in experiments on MMW biological effects at the cellular level. Finite-difference time-domain (FDTD) technique is employed to compute the 6 mm MMW power density (PD) irradiated into cells in a typical culture dish and the MMW power absorption density (PAD) of cells, followed by the qualitative and quantitative analyses on the outcomes. Indicated in the results, distributions of MMW PD irradiated into cells and the PAD of cells are complicated with evident inhomogeneity so that MMW dosimetry varies a lot in different position whose influence on the experimental outcomes is not neglectable. Consequently, the precise determination of dosimetry is of great importance to be conducted in related experiments. PMID- 15884547 TI - [Medical image automatic adjusting window and segmentation]. AB - Image guided surgical navigation system is the most advanced surgical apparatus, which develops most rapidly and has great application prospects in neurosurgery, orthopaedics, E.N.T. department etc. In current surgical navigation systems, windowing, segmenting and registration of medical images all depend on manual operation, and automation of image processing is urgently needed. This paper proposes the algorithm which realizes very well automatic windowing and segmentation of medical images: first, we analyze a lot of MRI and CT images and propose corresponding windowing algorithm according to their common features of intensity distribution. Experiments show that the effects of windowing of most MRI and CT images are optimized. Second, we propose the seed growing algorithm based on intensity connectivity,which can segment tumor and its boundary exactly by simply clicking the mouse, and control dynamically the results in real time. If computer memory permits, the algorithm can segment 3D images directly. Tests show that this function is able to shorten the time of surgical planning, lower the complexity, and improve the efficiency in navigation surgery. PMID- 15884548 TI - [Selection of peritoneal dialysis schemes based on multi-objective fuzzy pattern recognition]. AB - The fuzziness in the selection of peritoneal dialysis schemes is discussed. A method of multi-objective fuzzy pattern recognition and its application in the selection of peritoneal dialysis schemes are presented. The method was first applied to the field of peritoneal dialysis. The conclusion showed that this method agreed with doctors' opinions. It provides a new idea for research in this field. At the same time, since the method is simple and easy to use, it can be of wide application. PMID- 15884549 TI - [The Doppler fetal heart rate detection with wavelet transform]. AB - Various interference and noises should be eliminated while picking up the signal of fetal heart rate from the Doppler fetal signal in order to obtain smooth curve of the fetal heart Doppler signal and so to figure out the fetal heart rate. In this study, the mean frequency shift curve estimated from the Doppler fetal signals was smoothed using wavelet transform. The results showed that the fetal heart rate obtained by computing from the autocorrelation waveform based on the smoothed mean frequency shift curve was more accurate than that based on unsmoothed curve. PMID- 15884550 TI - [Synthetic effect analysis of heart rate variability and blood pressure variability on driving mental fatigue]. AB - Driving mental fatigue is a substantial problem and threatens people's health and life. This investigation was made to evaluate the driving mental fatigue based on power spectral analysis of heart rate variability (HRV) and blood pressure variability (BPV) synthetically. Twenty health male subjects (28.8 +/- 4.3 years) were required to perform the simulated driving for 90 min. The frequency domain indices of HRV such as low frequency (0.04-0.15 Hz, LF), high frequency (0.15-0.4 Hz, HF), LF/HF together with the indices of hemodynamics such as blood pressure (BP) and heart rate (HR) of the subjects were calculated and anlyzed after the simulated driving. The results showed that sympathetic activity of the subjects enhanced after the simulated driving while parasympathetic activities decreased. The sympathovagal balance was also improved. Remarkable linear correlations were observed between all the corresponding indices of HRV and BPV (P<0.05). The findings suggested that the evaluation of autonomic activities by means of multi variable synthetic effect analysis is scientific and objective. PMID- 15884551 TI - [Genotoxicity comparison between gasoline- and methanol-fueled exhaust by TK gene mutation assay]. AB - Methanol fuel is a most promising substitute for gasoline. It is scarcely reported about methanol-fueled exhaust on the health effect, neither about genotoxicity research between methanol- and gasoline-fueled exhaust. In the present study, the two kinds of exhaust were sampled directly from tailpipe at the same type bus, the same state, L5178Y thymidine kinase (TK) gene mutation assay was used to investigate their genotoxicity at the same dose range, and compared with micronucleus and comet assay. The results showed that the genotoxicity of gasoline-fueled exhaust is stronger than that of methanol-fueled exhaust, while the cytotoxicity of methanol-fueled exhaust is stronger than that of gasoline-fueled exhaust at dose range. The study demonstrated that L5178Y TK gene mutation assay is more sensitive than micronucleus and comet assay. PMID- 15884552 TI - [Fast extraction of iso-surface in surface rendering of medical images]. AB - In this paper a new direct volume rendering method is presented for fast extraction of iso-surface by adopting the idea from the Shear-Warp algorithm. By creating the sorted volumetric data from the original volume data and specifying a value range of data which determines the part of the sorted volumetric data traversed, the amount of volume data traversed would be reduced obviously and the extraction operation of iso-surface would be very fast. In addition, we can adjust the value range to obtain the different rendering speed and image quality according to the purpose in application. Moreover, the proposed algorithm will not output any intermediate data after the sorted volumetric data being produced. Therefore, it is possible to realize the rapid 3D surface reconstruction for medical images on the personal computer without the support of any hardware accelerator. PMID- 15884553 TI - [The mechanism and application of the correction for spherical myopia using small beam excimer laser]. AB - In the laser ablation refractive surgeries, the corneal ablation model and its realization by laser are difficult to design. They greatly affect the results of those surgeries. This paper firstly presents a mathematical model for corneal ablation to correct the refractive error of spherical myopia, and then gives a technique of planning laser focus positions on cornea to realize the ablation model. Based on the principle of the correction for spherical myopia using small beam laser, our excimer laser corneal surgery system has been able to perform such refractive surgery. Now the corneal surgery system has been applied to clinical practice. Thirty-nine eyes with spherical myopia underwent LASIK using this kind of system. Their refractive states have been greatly improved. Preoperatively, the mean sphere was -5.57+/-2.95 D and the mean uncorrected visual acuity 0.12+/-0.07. One day after surgery, the mean sphere was -0.03+/ 0.57 D and the mean uncorrected visual acuity 0.90+/-0.29. One month after surgery, the mean sphere was -0.68+/-0.98 D, and the mean uncorrected visual acuity 1.0+/-0.26. PMID- 15884554 TI - [A novel ROI extracting technique based on wavelet transform for the detection of micro-calcifications in mammograms]. AB - In order to preprocess mammograms for diagnosing the early cases of breast cancer and improving the computational efficiency in the computer-aided detection of micro-calcifications in mammograms, we have advanced a novel processing technique for the extraction of micro-calcification region of interest (MROI). The proposed method is based on a three-step procedure: (1) the mammogram is divided into sub images of the same size; (2) the wavelet multi-resolution method is conducted on the sub-images, and the parameters related to wavelet transform and threshold T are discussed according to rho; (3) the classification of sub-images is determined by T. It is tested with 20 mammograms and the results show that the method can achieve a true positive rate as high as 89.7% with a false positive rate as low as 2.1%. PMID- 15884555 TI - [Measurement of otoacoustic emissions by sound card]. AB - A new method in OAE's measurement by computer sound card is introduced. It is a measurement system using SF-1 detector. We designed the appropriate analog filter amplifier peripheral, utilized the mature technology of computer sound card, and recorded the OAE signal using Windows' API function programming by VB. The recorded signals can be analyzed by Matlab, such as digital filtering and Coherence average. The system is, in the main, cost-effective. The statistical results of the experiments proved the reliability of the method. PMID- 15884556 TI - [Analysis of nonlinear behavior in cardiac QRS-waves integral]. AB - QRS-waves integral can reflect the activities of ventricular depolarization. In this paper are analyzed the nonlinear characters in QRS-waves integral: reconstructing state space with these integral signals after nonlinear noise reduction, calculating its attractor dimension and the largest Lyapunov exponents. With the results based on MIT-BIH database we proposed that QRS-waves integral is of chaos nature. When compased with HRV nonlinear analysis, QRS-waves integral analysis was found to be of more advantages in some cases such as premature ventricular contraction. This QRS-waves integral nonlinear analysis may be of use in evaluating clinical cardiac function. PMID- 15884557 TI - [Primary study on the method for separating mature sperms from human testicular tissue]. AB - This study was aimed to explore a simple and applicable method of separating mature sperms from human testicular tissue for intracytoplasmic sperm injection and embryo transfer. The suspension of human testicular tissue was cultured in 10% human serum albumin and human tubule fluid with different concentrations (0 u/ml; 50 u/ml; 100 u/ml; 150 u/ml; 200 u/ml) of hyaluronidase for 24 h, and then the Percoll gradient centrifugation was processed to separate the sperms; meanwhile the sperms were counted and graded according to their motility. The difference in quality and quantity among the groups and the difference between the groups and the zero-hour culturing group were detected. It was shown that the four hyaluronidase-treated groups contained large quantity and high quality of sperms as compared with the two contrast groups (P<0.01). The groups in the solution of 50 u/ml, 100 u/ml and 150 u/ml concentrations of hyaluronidase had almost the same amount of sperms that displayed higher motility as compared against the sperms in the group treated with 200 u/ml concentration of hyaluronidase (P<0.01). There was no difference between the two contrast groups (P>0.05), or among the groups treated with 50 u/ml, 100 u/ml, and 150 u/ml of hyaluronidase concentration (P>0.05). This method of adopting hyaluronidase with Percoll gradient centrifugation in the process for separating mature sperms from human testicular tissue is applicable. It can increase the quantity and quality of sperms separated from testicular tissue suspensions when adequate concentrations of hyaluronidase is used. PMID- 15884558 TI - [Receptor selection and B cell immune tolerance]. AB - The immune system of mammalian has developed sophisticated mechanism to deal with diverse unknown antigens by random rearrangement of V, D and J antigen gene fragments. The immune self-tolerance is the mechanism to avoid self-destruction by the gene rearrangement generated autoreactive lymphocytes. Until recently it was believed that autoreactive lymphocytes are either deleted or rendered unable to respond by the supposed cell or clone selection mechanism. However, recent findings from a number of laboratories suggest instead of cell selection but receptor selection plays an essential role in immune self-tolerance. Receptor selection is carried out by secondary or nested rearrangement of antigen receptor gene fragments, and can occur at different stages of lymphocyte differentiation. Furthermore, secondary rearrangement of receptor gene also plays an important role in shaping immune response after the interaction of receptor with antigen by altering its specificity. PMID- 15884559 TI - [The application and development of ultrasonic scalpel]. AB - Ultrasonic scalpel has the advantages of less bleeding, less damage to surrounding tissue, early healing, etc. The applications of ultrasound in medical surgery include incision and hemostasis, phacoemulsification, tumor aspiration, fat aspiration, and bone cutting. In this paper, the principles, development, characteristic, key technology and clinical application of different ultrasonic scalpel in the world are introduced, and the domestic application and future development of ultrasonic scalpel are prospected. PMID- 15884560 TI - [Progress in researches on the synthesis of poly (propylene fumarate) and its crosslinking characteristics]. AB - Poly (propylene fumarate) is an unsaturated linear polyester, which degrades, in the presence of water, into propylene glycol and fumaric acid, and the degradation products can be cleared from the human body by normal metabolic processes. In this paper, different methods of the synthesis of PPF are listed, the crosslinking characteristics of polymerizing composite as well as the mechanical properties of cross-linked composite are discussed, and the potentialities of PPF composite used as particularly degradable polymeric cement for orthopedic application are reviewed. PMID- 15884561 TI - [Research progress in the technique of integrating multiple imaging modalities in the functional brain imaging]. AB - In the last few years, brain science has exhibited an explosive growth in hemodynamic/metabolic data on brain function. In particular, PET and functional MRI provide excellent spatial resolution, but their temporal resolutions are severely limited by relatively slow responses of brain metabolism. By contrast, EEG and MEG are able to track modulations of current source activity at millisecond time scales, but suffer from poor spatial resolution. Hence, high resolution spatiotemporal imaging of brain activity requires integration of information from multiple signal modalities. In this paper we discuss what is known about the physical and physiological basis of the imaging signals, suggest various approaches that have been proposed for integrating information across multiple modalities, and briefly analyze the future directions and remaining challenges in the field of multi-modality integration. PMID- 15884562 TI - [Research progress of the drug-coated stents]. AB - Emplacing a stent in the coronary artery has many characteristics. The installation is easy to do and the stent has evident curative effect. However, it will cause thrombosis and immunoreaction because it is metal. So suburgent thrombosis and restenosis after surgeries are still two major complications. The drug-coated stent is one kind of the drug-eluting stents, whose metal surface is coated by some polymer that combines with a sort of effective drug or antibody. It can transport the drug or antibody to the localily of pathological changes and there in it improves the local drug concentration. In this paper the research progress of interrelated issues about the drug-coated stent is reviewed. PMID- 15884563 TI - [Design, production and clinical application of custom total hip prosthesis]. AB - The key to the research of the hip prosthesis is to prolong the effective using time of the hip prosthesis, but bone loss and aseptic loosening are the influential factors in this aspect. Since each person has his own characteristic, it is difficult for the standard hip prosthesis to fit with the patient's femoral canal, so the stability can not the ensured. Moreover, some patients have congenital abnormality or a serious defect in bone or arthrosis, which makes an obvious deviation from the normal, therefore they can not use the standard hip prosthesis. CAD/CAM is useful for to individual three-dimensional femoral reconstruction, which can improve the stability of the prosthesis and effectively prevent the loosening of prosthesis. This article introduces the advantage, design, production and the problems of custom CAD/CAM hip prosthesis. PMID- 15884564 TI - [Development of hypothyroidism therapy with thyroid hormone]. AB - The management of patients with primary hypothyroidism is straightforward. Recent studies suggest, however, that standard thyroid replacement therapy with thyroxine may not be completely effective in relieving the symptoms of hypothyroidism, and that there may be a role for combined use of thyroxine and triiodothyronine (T3) in the replacement therapy. Furthermore, animal studies suggest that the direct contribution by the thyroid to circulating T3 concentrations may be important, and that thyroxine alone may not be adequate treatment for hypothyroidism. In studies on thyroidectomized rats, it was found that the achievement of normal tissue concentrations of T3 required either the thyroxine at high doses which resulted in the suppression of TSH secretion, or the combined thyroxine/T3 treatment, which was able to normalize the serum thyroxine, T3 and the TSH concentrations, and the levels of thyroxine and T3 in most peripheral tissues. If the same is true of humans, there might be a more physiological replacement regimen for hypothyroid patients other than the replacement regiment of using thyroxine alone. PMID- 15884565 TI - [The mechanotransduction mechanism of how osteoblasts respond to mechanical stimulation]. AB - The stress environment regulates the factors of growth, resorption and remolding in bone tissue. Mechanical stimulation at cell physical level affects the physiological activity of osteoblasts, including proliferation, ALP activity and osteocalcin production. Mechanotransduction is a procedure which transduces the biophysical force into biochemical responses. It is also the basis of many physiological functions. The early response genes (c-fos, c-jun), the second message systems (Ca2+, NO, cAMP) and the mechano-sensitive cation channel are involved in the mechanotransduction course when osteoblasts respond to the mechanical stimulation. PMID- 15884566 TI - [Ultrasound-mediated gene delivery]. AB - Recently there have been increasing interests in ultrasound-mediated gene delivery techniques. This paper reviews the general concept of sonoporation, the effect of contrast agents, and the in vitro and in vivo applications of such techniques. Factors which contribute to the efficiency of gene delivery were also discussed. The technology is promising and has great potential in targeted drug delivery and gene delivery applications. PMID- 15884567 TI - [Individualized energy metabolic monitoring and its clinical application]. AB - The nutritional status affects the factors of patient outcome such as the therapeutic intervention, the length of hospitalization and prognosis. Personalization of nutritional support treatments can minimize the complications that may arise from overfeeding and underfeeding. Appropriate nutritional support requires an individual nutrition assessment based on each patient's caloric and regularly monitored respiratory quotient to accommodate continuing changes in the patient's condition. PMID- 15884568 TI - [Current researches in ultrasonic angioplasty]. AB - Low frequency high power ultrasound is a recent addition to the list of methods for arterial angioplasty. This paper introduced the effects of the low frequency high power ultrasound and summarized the results of many experiments with its use for different kinds of arterial obstruction. PMID- 15884569 TI - [Advances in apoptosis induced by biomaterials]. AB - Biomedical materials are the biomaterials that, used in physiological system for diagnosis, treatment, plerosis or replacement of tissues and organs. Apoptosis, also known as PCD or ACD, is a normal physiological mechanism of cell in organism and a process of automatic cell death in which multicell organism modulates the development of organism and maintains the stability of internal environment. The human beings are able to understand the interaction between the material and organism at the molecular level due to the widely-used biomedical material and the development of material science, life science and biological technology. The research of that interaction is mainly focused on biocompatibility, while much attention has been drawn to the apoptosis induced by biomaterial concerning that apoptosis could be caused by inducing factor, and many therapies of diseases are closely related to inducing apoptosis. Based on the recent research advances of apoptosis in life science and the development of biomaterials, the pathways of apoptosis induced by biomaterials were reviewed; from the different views, the pathways of signal transduction of apoptosis include traditional pathway of signal transduction, the pathway of death receptor, and the pathway through mitochondrion. By the other way, the pathways of apoptosis caused by reactive oxygen species induced by biomaterials and apoptosis by affecting cell adhesion to biomaterials and so forth were discussed also. It indicates that the pathways to apoptosis due to biomaterials possess the characteristics of variety, intercrossing and multiplicity. It is essential for a research to inquire into the mechanism of apoptosis that is induced by biomaterials, and further into the manufacturing of biomaterials. This review is devoted to shedding light on the wide application of biomaterials in the therapy of human diseases, especially in the therapy of cancer that is closely related to apoptosis. PMID- 15884570 TI - [Advances of study on the relationship between vascular endothelial growth factor and ischemic cerebral vascular disease]. AB - Vascular endothelial growth factor is a potent inducer of angiogenesis identified in recent years, and its relationship with ischemic cerebral vascular diseases is a hotspot for many investigators at present. In this review, we focus on the recent evidence for VEGF expression, biological effect and its role in the treatment and prognostication of ischemic cerebral vascular disease. PMID- 15884571 TI - [The advance of experimental cell division biomechanics]. AB - The advance of experimental cell division and proliferation in the field of cell biomechanics is presented in this paper. The emphasis is placed on the research in the mechanics mechanism of cleavage furrow and in the measurement of constricting force about cleavage furrow. PMID- 15884572 TI - [Characterization of anticoagulant biomaterial and its development]. AB - Good anticoagulant biomaterials need good surface chemical properties, good mechanics performances and particularly good characteristics of biocompatibility, including tissue compatibility and hemocompatibility. In order to understand with greater clearness the anticoagulant biomaterial, we have to characterize them by different methods. In this paper, the approaches to assessing and displaying the characteristics of anticoagulant biomaterial are reviewed in three aspects, namely the surface chemical properties and structure, the mechanics performances the and the biocompatibility of anticoagulant biomaterial. PMID- 15884573 TI - Osteotomes vs. traditional drilling for implant placement. PMID- 15884574 TI - The role of official veterinary services in dealing with new social challenges: animal health and protection, food safety, and the environment. AB - The authors describe the role of Veterinary Services in dealing with developments in society and the challenges they present. In an increasingly globalised world, in which there are new methods of production, a new and changing relationship between humans and animals and increasingly complex and global problems, the protection of public health and the safety of the environment in which people and animals live require new skills, new knowledge and new technical-scientific responses. Official Veterinary Services are responsible for maintaining hygiene and animal health standards in primary and secondary production and ensuring fair trade. To achieve this, Veterinary Services must ensure that their organisational structures meet internationally recognised quality assurance standards. PMID- 15884575 TI - [The requirements of society in terms of the safety of food of animal origin--a reflection on the case of France]. AB - Are consumers obsessed by 'zero risk'? Is their relationship to food of animal origin irrational? The comments and reactions during the bovine spongiform encephalopathy (BSE) crisis that began in November 2000 certainly suggest so. However, sociological surveys conducted at the height of the crisis refute this impression: judging by individual perceptions, risk is not an obsession; it is only one of several aspects of people's relationship to animal products. Furthermore, concern about healthy eating is not new and, regarding meat specifically, it is linked to the fundamental ambivalence of our relationship to animal flesh. Taking these invariants into account, this paper aims to identify the specific characteristics of our contemporary societies that cause us to set requirements in terms of the safety of products of animal origin. We adopt the thesis of the vulnerability of our societies: a combination of various factors triggers crisis situations, the scale of which is often out of proportion to the gravity of the danger. Vulnerability is particularly acute with respect to food, because the volatility of consumer behaviour is at odds with the relative inflexibility of the livestock industry. Consequently, it is imperative to put an end to certain misconceptions, particularly the requirement of 'zero risk'. The requirements of our societies must be expressed in procedural ratherthan substantial terms. This leads us to explore the characteristics of a new system of risk management, based on the idea of the 'responsible consumer', who participates in decision-making. PMID- 15884576 TI - [The precautionary principle and its legal implications in the area of food safety]. AB - The concept of the 'precautionary principle' is not independent of the general principles of risk assessment and cannot be used as a justification for circumventing the legal obligations established at international level by World Trade Organization (WTO) agreements. On the contrary, the principle, which allows policy-makers to act without waiting until all the relevant scientific information is available, is included in the discipline established by the WTO Agreement on the Application of Sanitary and Phytosanitary Measures (SPS Agreement) and confirmed by jurisprudence. Sanitary measures adopted provisionally in a context of scientific uncertainty should nevertheless endeavour to comply with the general principles of risk management established by the SPS Agreement, i.e. they should be appropriate, non-discriminatory, consistent and not more trade-restrictive than required. The Codex Alimentarius has been working for several years to develop a set of principles for the application of risk assessment, including the precautionary principle when information is insufficient or incomplete. These principles will serve as a guide first for the work of the Codex and subsequently for governments. The European Union has made provision for the application of the precautionary principle, in accordance with circumstances, in Community food legislation, while remaining in compliance with the legal obligations stemming from the SPS Agreement. PMID- 15884577 TI - Risk analysis: assessment, management and communication. AB - Import risk analysis is now a well-established discipline which aims to assist Veterinary Services to answer the following questions: 'What can go wrong?', 'How likely is it to go wrong?', 'What would be the consequences of it going wrong?' and 'What can be done to reduce either the likelihood or the consequences of it going wrong?'. Risk communication is that part of the overall process which, among other things, helps the decision-maker to determine whether a particular risk is acceptable or not. Good risk assessment and communication are dependent on clear formulation of the question to be answered. Scenario trees and influence diagrams are very useful tools in assessing and communicating risk. The authors outline the import risk analysis procedures adopted by the Veterinary Services of one Member Country of the OIE (World organisation for animal health). PMID- 15884578 TI - The organisation of federal Veterinary Services in Canada: the Canadian Food Inspection Agency. AB - The organisational design of a national Veterinary Service is critical to the overall quality and integrity of its animal health and veterinary public health infrastructure. It is well recognised that the diversity of political, economic and social situations which exist in and between countries dictates that no one model of organisational structure can be applied to all circumstances. In Canada, a re-organisation of the approach of the federal government to food inspection in 1997 resulted in the transfer of the veterinary administration to a newly created agency called the Canadian Food Inspection Agency (CFIA). The authors provide a short background on the impetus for the creation of the CFIA and an overview of its organisational structure and responsibilities in animal and veterinary public health and food safety. Also included are the logic models that were developed for the federal Veterinary Services as part of their quality and performance management framework. Integrating all federally mandated food inspection systems under the CFIA has had concrete benefits in clarifying roles and responsibilities, reducing overlap and duplication of programme functions, improving service delivery and facilitating federal-provincial collaboration. Moreover, the strength of the organisation lies in the ability of the Canadian Veterinary Services to adhere to the fundamental principles of quality which are recommended by the OIE (World organisation for animal health) for the evaluation of Veterinary Services. No single organisational structure can guarantee a highly effective or competent Veterinary Service. Common challenges exist that may or may not be addressed in whole or in part by the organisational structure. The challenges highlighted in this paper provide further thoughts on the management of shared jurisdiction, meeting public health objectives, balancing science and political accountability, and defining the role and jurisdiction of veterinarians. PMID- 15884579 TI - The federal system of Veterinary Services in Switzerland. AB - The Veterinary Service is described in the context of the federal state system in Switzerland. The Federal Veterinary Office (FVO) plays a leading role in this Service. Apart from its responsibility for legislation, the most important task of the FVO is the dissemination of information throughout Switzerland. The responsibility for ensuring that the laws are implemented lies with the 26 cantons. Veterinary public health is a key function of the Veterinary Service, where the core focus is on the implementation of the 'stable to table' concept. Animal protection is also an important function. A certified Veterinary Service requires a high-quality system based on professionalism, objectivity and evaluation. In this paper, the various strengths and weaknesses of the system in Switzerland are discussed and the disadvantages are analysed in depth. PMID- 15884580 TI - [The improvement of the animal health risk analysis system in France]. AB - After several decades of gradual improvement in its system for managing health risks, France was confronted in 1996 with the bovine spongiform encephalopathy crisis. This triggered a collective questioning, which highlighted the need to reform a system that had shown its limitations. Risk analysis, established as a key principle by the Agreement on the Application of Sanitary and Phytosanitary Measures (SPS Agreement) of the World Trade Organization (WTO), was cast as the necessary basis of the reform, objectives of which were to better identify priority risks in order to ensure the protection of human and animal health, and to improve the quality of measures implemented by the public authorities. The Act of 1 July 1998 founded several independent risk assessment agencies, including the French Agency for Food Safety (AFSSA), with the specific mandate of food safety at every stage of the food chain. Other organisational reforms enhanced the new system, notably the separation between the functions of risk management and economic support for food industries, initially at central level, then in 2002 at the level of field services in the 100 French departements. Lastly, new procedures were introduced. These were designed, in accordance with the principles of risk assessment, to better identify and to individualise the different decision-making sequences. The decision-making process was extended to include submission to the agency in charge of evaluating health risks and examination by the agency of the resulting draft decision. PMID- 15884581 TI - [The organisation and future development of Veterinary Services in Latin America]. AB - Latin America undoubtedly has comparative advantages in the fields of animal production, animal health and the production of food of animal origin. However, countries in Latin America must build on these strengths if the continent is to become more competitive and be able to deal with the complexities of world markets. To do this, Veterinary Services must define their objectives and establish quality standards on which to base their work. For this to occur, the State must create well-defined regulations, establish systems of audit and find ways of working which allow for a high degree of coordination and collaboration between the public and private sectors. This should be done within a framework of a quality assurance system, which allows for responsible accreditation and independent audit and evaluation. The author discusses the approaches of the different countries in the region to animal health, zoonosis, food safety, veterinary drugs control, animal welfare and export-import control. All programmes relating to these issues must be based on technical information gained through epidemiological surveillance, the network of diagnostic laboratories, quarantine systems, risk analysis, identification and traceability of animals and animal products, registration and control of veterinary drugs, and food safety research. In some countries these systems are already being developed. Maintaining good international relations and cooperating with neighbouring countries is always a challenge for official Veterinary Services and international organisations such as the OIE (World organisation for animal health) have a key role to play in facilitating these relationships. PMID- 15884582 TI - Veterinary Services in Belize: adapting organisational models to the needs of small economies. AB - International trade in agricultural goods is governed by the Agreement on the Application of Sanitary and Phytosanitary Measures (the 'SPS Agreement') of the World Trade Organization. Small economies, such as the economy of Belize, must have the necessary infrastructure in place to meet their obligations and exercise their rights under the SPS Agreement. In response to the shortcomings of a small economy, the Belize Government has established the Belize Agricultural Health Authority (BAHA) as its organisational model, to meet the challenges of providing Veterinary Services in this new millennium, as well as the demands of the international trade in animals and plants and their products. This model became operational in the year 2000 and has received national and international recognition as an appropriate model for small economies. The success of BAHA is due to strong political support and its dual public and private sector characteristics. PMID- 15884583 TI - [The present and future organisation of Veterinary Services in Africa]. AB - In Africa, for many people, breeding animals is at the heart of their culture. For the poorest families animals play a vital role in most aspects of daily life, providing food, income and a place in society. Moreover, for poor farmers, livestock are the main source of energy, as they are used for farming and for the transport of their products. Unfortunately, trans-border animal diseases remain, to this day, one of the main obstacles to the development of breeding in Africa. They bring about heavy losses, direct and indirect, to national herds and often cause public health problems as well. Regional and international trade in animals and animal products is hampered due to these diseases, which are also one of the main causes of poor performance in the breeding sub-sector and of the growing gap between supply and demand of meat and milk. The challenge of maintaining sustainable and competitive growth in animal production requires major improvements in production methods and animal health. Better animal health protection in turn, depends on national policies which confer upon Veterinary Services the responsabilities and organisational structures they once had, but which unfortunately have been withdrawn by structural adjustment policies. PMID- 15884584 TI - The organisation of the Department of Veterinary Services in Malaysia. AB - The Department of Veterinary Services (DVS) in Malaysia was established in 1888 as an agency to control exotic and domestic animal diseases. Over the years, the structure and functions of the organisation have evolved to meet the growing demand for veterinary services. The responsibilities of the Veterinary Services are enshrined in the Constitution of Malaysia. The current organisation of the DVS is structured to achieve the following objectives:---to prevent, control and eradicate animal and zoonotic diseases--to facilitate the growth and development of a strong animal industry--to ensure that animal products for human consumption are wholesome, clean, safe and suitable to be consumed--to facilitate the growth and development of the animal feed industry--to ensure the welfare and well-being of all animals. To meet these objectives the DVS has nine different divisions, as follows: Planning and Evaluation, Epidemiology and Veterinary Medicine, Veterinary Public Health, Research and Development, Industry Development, Production and Development of Genetic Resources, Human Resource Development (HRD), Enforcement, and Administration. The development of the animal industry is managed through national development policies, including the Third National Agriculture Policy. The basis for current programmes for disease control and animal industry development is the Eighth Development Plan (2001-2005). Over the period of this Plan, Malaysia will address the need for sanitary and phytosanitary measures by developing specific programmes covering all fields of the animal industry. This is just one way in which Malaysia is meeting the challenges of the increased liberalisation of trade created by the World Trade Organization and the Association of Southeast Asian Nations Free Trade Area. The development of the industry is focused on the major commodities, namely, beef, mutton, poultry meat, eggs, pork and milk. Other commodities receive support if it is considered economically viable. All support services are being strengthened, particularly the HRD division. The organisation and functions of the DVS are constantly being reviewed in accordance with changes in the animal industry and the nature of the services in demand. PMID- 15884585 TI - The present and future organisation of Veterinary Services in Asia: the examples of the Republic of Korea and Japan. AB - The Veterinary Services of the Republic of Korea and Japan are reviewed and summarised. To cope with ever-increasing demands on Veterinary Services, both countries have made continued efforts to expand the services, which currently only have a limited number of government personnel. To overcome some of the problems associated with international trade, e.g. outbreaks of foot and mouth disease and classical swine fever (hog cholera), the authorities of the Republic of Korea are considering strengthening quarantine services by combining the National Veterinary Research and Quarantine Service and the National Plant Quarantine Service of the Ministry of Agriculture and Forestry (MAF). In the field of veterinary public health services, the Republic of Korea and Japan have different organisational structures. All Veterinary Services, including veterinary public health, are kept within the MAF in the Republic of Korea, whereas in Japan, the Ministry of Agriculture, Forestry and Fisheries (MAFF) deals with general animal production and health services and the Ministry of Health, Labour and Welfare (MHLW) deals with the safety of animal products, food poisoning and the prevention/control of zoonoses, such as rabies and Q fever. In Japan, after the occurrence of bovine spongiform encephalopathy in 2001, the authorities undertook a thorough review of the Veterinary Services, focusing particularly on food safety. A reorganisation of the administrative structure was proposed, and will be completed by the end of 2003. Subject to the approval of the Diet (House of Councillors), an independent 'Food Safety Commission' will be created within the cabinet office which will undertake risk analyses related to foods, and risk management will be carried out by the relevant ministry, i.e. the MAFF or the MHLW. The Animal Health Division of the MAFF will also be moved from the Livestock Industry Department to a new 'Consumer Safety Department' of the MAFF, where additional responsibilities in fish health control and feed safety control will be assumed. PMID- 15884586 TI - The organisation of the State Food and Veterinary Service of Lithuania. AB - The author describes the State Food and Veterinary Service of Lithuania, the competent authority for public and animal health in Lithuania. The Director of the Service, the Chief Veterinary Officer, reports directly to the Prime Minister. Lithuania is divided into ten counties and thirty-four districts. The State Food and Veterinary Service has regional units in all the counties and districts and has a staff of 1,380 employees. The main tasks of the State Food and Veterinary Service are to implement the basic EU principles of market supervision in Lithuania, with an emphasis on ensuring food safety and control; to establish a unified food control system following the principle "from farm to fork"; to create a favourable environment for the free movement of goods between Lithuania and the countries of the EU while ensuring the safety of imported products of animal origin; to promote continuous improvements in the welfare of animals and to protect the animal population from contagious diseases. PMID- 15884587 TI - How to promote joint participation of the public and private sectors in the organisation of animal health programmes. AB - It is generally accepted that the first recorded outbreaks of foot and mouth disease (FMD) in South America occurred around 1870. The disease emerged almost simultaneously in the province of Buenos Aires (Argentina), in the central region of Chile, in Uruguay and in southern Brazil, due to the introduction of livestock from Europe. Argentina set up an agency for the control and eradication of FMD in 1961, Brazil began disease-control activities in Rio Grande do Sul in 1965, Paraguay and Uruguay initiated similar programmes in 1967, Chile in 1970 and Colombia in 1972. A common characteristic was observed in all early national FMD programmes, namely, they were developed, financed, operated and evaluated by the public sector, without major participation from the private sector, except when buying vaccines and abiding by the regulations. In 1987, the Hemispheric Foot and Mouth Disease Eradication Plan (PHEFA: Plan Hemisferico para la Erradicacion de la Fiebre Aftosa) was launched and the private sector played a prominent role in achieving the eradication and control of FMD in several countries. However, this model of co-participation between the public and private sectors has suffered setbacks and a new approach is being developed to find ways in which local structures and activities can be self-sustaining. PMID- 15884588 TI - [Collaboration between official veterinarians, private veterinarians and livestock producer organisations]. AB - In order to guarantee that only healthy animals and safe animal products are placed on the market, official veterinarians, private veterinarians and livestock producer organisations must work together within a single structure that reports to a single centralised authority. Each party must participate in developing and implementing programmes, in financing these programmes and in taking responsibility for the results (joint decision-making, joint financing, joint accountability). The author describes how this mode of collaboration and integration between the three parties operates, and explains their duties and obligations. PMID- 15884589 TI - Veterinary diagnostic laboratories and their support role for Veterinary Services. AB - National Veterinary Services administer a number of regulatory programmes, such as foreign animal disease (FAD) surveillance and exclusion of FADs, and certification of regions as free from disease. Laboratory testing is an important part of any surveillance or control programme. Most countries have a national laboratory which performs testing for FADs and provides support for national disease eradication and control programmes. State laboratories provide testing for surveillance programmes and export purposes, in addition to diagnosis of clinical cases. Many national and state laboratories are developing quality assurance programmes to assure the reliability of testing results. Veterinary Services are reliant on the diagnostic expertise of the laboratory system of that country to be able to respond to FAD introductions and to provide the surveillance programmes needed to detect the introduction of diseases and to certify freedom from disease. PMID- 15884590 TI - [The role of reference laboratories in animal health programmes in South America]. AB - The contribution of the Panamerican Foot and Mouth Disease (FMD) Centre (PANAFTOSA), as an OIE (World organisation for animal health) regional reference laboratory for the diagnosis of FMD and vesicular stomatitis, and for the control of the FMD vaccine, has been of fundamental importance to the development, implementation and harmonisation of modern laboratory procedures in South America. The significance of the work conducted by PANAFTOSA is particularly obvious when one considers the two pillars on which eradication programmes are based, namely: a well-structured regional laboratory network, and the creation of a system which allows technology and new developments to be transferred to Member Countries as quickly and efficiently as possible. Over the past decade, PANAFTOSA has kept pace with the changing epidemiological situation on the continent, and with developments in the international political and economical situation. This has involved the strengthening of quality policies, and the elaboration and implementation of diagnostic tools that make for more thorough epidemiological analyses. The integration of PANAFTOSA into the network of national laboratories and its cooperation with technical and scientific institutes, universities and the private sector means that local needs can be met, thanks to the design and rapid implementation of methodological tools which are validated using internationally accepted criteria. This collaboration, which ensures harmonisation of laboratory tests and enhances the quality of national Veterinary Services, serves to promote greater equity, a prerequisite for regional eradication strategies and this in turn, helps to increase competitiveness in the region. PMID- 15884591 TI - The obligations of Member Countries of the OIE (World Organisation for Animal Health) in the Organisation of Veterinary Services. AB - The authors discuss the mission, organisation and resources of Veterinary Services in the new international trading environment and examine how the standards for Veterinary Services, contained in the OIE (World Organisation for Animal Health) International Animal Health Code (the Code), help provide the necessary support for Veterinary Services to meet their rights and obligations under the provisions of the Sanitary and Phytosanitary (SPS) Agreement of the World Trade Organization (WTO). The authors describe the challenges of gaining access to international trading markets through surveillance and control of OIE listed diseases. Finally, the approach in the Code to the principles underpinning the quality of Veterinary Services and to guidelines for evaluating Veterinary Services, is discussed. PMID- 15884592 TI - [Guidelines of the OIE (world organisation for animal health) for the organisation of Veterinary Services and their relations with private-sector partners, with particular reference to developing countries]. AB - Developing countries are being confronted with changes in the international rules governing trade in animal products. At the same time, state control over Veterinary Services, which has been in place for many years, is gradually giving way to private-sector involvement. In light of these developments, the International Animal Health Code (the Code), must ensure that it includes guidelines that take into account the new role of private-sector professionals. Integrating these professionals into the national networks of Veterinary Services is vital for the improvement of the services. Moreover, the situation in the field in developing countries has at times given rise to the emergence of 'alternative' methods of veterinary care, whose uncontrolled development may be an obstacle to achieving the required levels of service provision. It is recommended that countries shun apparently attractive short-term solutions, and adapt their national legislation to allow for the gradual reform of their Veterinary Services, pursuant to international standards. PMID- 15884593 TI - [Human resource programming and management: the experience of the French Veterinary Services]. AB - France has been endeavouring to come up with veterinary activity indicators by which measure the work of public Veterinary Services and to rationalise the geographical distribution of staff and financial ressources on the basis of objective criteria. An analysis of activities in 1998, and of new missions, has made it possible to: define criteria against which the tasks accomplished by the various divisions can be measured; prepare a method for establishing optimal distribution criteria for the human resources available; propose, for each departement, the allocation of human resources based on national distribution criteria but taking into account the individual needs of different areas; measure variations in activity resulting from new or modified missions, with a priori theoretical assessments (impact studies), and a posteriori control. In each department, the Direction generale de I'alimentation (DGAL) has adopted an ongoing management tool composed of: a national database quantifying the activities of its services in real time, a simulation tool, to calculate the human resources needs for the local units. PMID- 15884594 TI - [The role of a specialised risk analysis group in the Veterinary Services of a developing country]. AB - Since the World Trade Organization (WTO) Agreement on the Application of Sanitary and Phytosanitary Measures was established, risk analysis in trade, and ultimately in Veterinary and Animal Health Services, has become strategically important. Irrespective of their concept (discipline, approach, method, process), all types of risk analysis in trade involve four periods or phases:--risk identification-- risk assessment--risk management--risk information or communication. All veterinarians involved in a risk analysis unit must have in depth knowledge of statistics and the epidemiology of transmissible diseases, as well as a basic knowledge of veterinary science, economics, mathematics, data processing and social communication, to enable them to work with professionals in these disciplines. Many developing countries do not have enough well-qualified professionnals in these areas to support a risk analysis unit. This will need to be rectified by seeking strategic alliances with other public or private sectors that will provide the required support to run the unit properly. Due to the special nature of its risk analysis functions, its role in supporting decision making, and the criteria of independence and transparency that are so crucial to its operations, the hierarchical position of the risk analysis unit should be close to the top management of the Veterinary Service. Due to the shortage of personnel in developing countries with the required training and scientific and technical qualifications, countries with organisations responsible for both animal and plant health protection would be advised to set up integrated plant and animal risk analysis units. In addition, these units could take charge of all activities relating to WTO agreements and regional agreements on animal and plant health management. PMID- 15884595 TI - The implementation of a quality assurance procedure for the Veterinary Services of France. AB - Due to the increasing complexity of food production systems and the concerns that these systems raise, there has been increasing demand from the general public for more State control of these processes. In France, it is the official Veterinary Services who are responsible for food safety and who must respond to these demands. The Veterinary Service is formulating a quality assurance procedure in accordance with standard EN 45004-ISO 17020, which determines the requirements that inspection bodies must follow to be recognised, at national, European and international level, as competent and reliable. As part of this procedure, the Veterinary Service will review requirements in terms of organisation, functions, qualifications and resources. The progress of inspection service orders, from their conception by the Central Administration, to their implementation by decentralised services, must be carefully managed. It is essential that service orders be implemented effectively and systematically by using recognised methods and issuing adequate inspection reports. The training and qualifications of inspectors are very important: their skills must remain up-to-date so that there is always a network of qualified staff, that is, staff who have an understanding of production processes and who have recognised competences in terms of initial training, continuous professional development and adequate experience. The quality systems implemented will only meet expectations if they are continuously monitored by means of regular evaluations. For this reason, both internal and external audits are performed. These new practices contribute to establishing a basis for the improvement of internal evaluation. In order to facilitate the implementation of a quality assurance procedure for inspection services, several tools, that are linked with the information system of the government department responsible for food, are, or will be, at the disposal of the decentralised Veterinary Services, i.e. a national database, mail and service order processing software, and inspection procedures. PMID- 15884596 TI - [Veterinary Services: how to establish quality assurance in developing countries]. AB - The authors put forward a methodological proposal for introducing quality assurance into animal health service provision in developing countries, based on the experience of Brazil. This proposal is based on three key elements. The first and most important element is the continuing development of those involved in the process, not only from the technical and administrative standpoint (know-how), but also in terms of modelling new forms of behaviour and attitudes for the provision of services (social and personal skills). This requires a culture of ethical values, motivation and team working to ensure that those involved in the process become committed, highly effective employees who identify with the Veterinary Service's objectives as if they were their own. The second key element of this methodology is to consider the various health services to be provided as processes and to treat each of these services as a set of activities, inputs and procedures that are systematically organised for delivering the specified animal health service. Working with processes not only makes it possible to properly plan each service, it also facilitates the identification, analysis and effective resolution of any problems that may arise while the service is being delivered. The third key element of the proposed methodology is the constant control and improvement of the processes making up the various health services, in particular using the Deming cycle (plan--do--check--act). The proposed strategy calls for realism and pragmatism in order to avoid creating false expectations by developing over-ambitious quality systems. It is advisable to avoid major publicity at the start of activities and to create well-defined, localised improvement programmes with anticipated results that are simple, direct and discernible to any observer within a short time frame. In the early stages, the term "quality" needs to be replaced with "better service delivery". It is necessary to ensure that staff is given some type of acknowledgement or feedback when positive results are achieved. Great emphasis must be placed on staff motivation and awareness-raising activities. PMID- 15884597 TI - [The contribution of quality assurance to national veterinary organizations in developing countries: the case of Africa]. AB - Food safety has, within a few years, become an essential concern for producers and consumers, especially in Western countries, but also in Africa. African countries are increasingly aware of the issues involved, and of the need to react and to implement appropriate strategies in order to avoid public health risks and obstacles to their export markets. Most countries recognise that the overall sanitary quality of foodstuffs produced and distributed in the region must be improved. The OIE (World organisation for animal health) recommends that Veterinary Services attempt to improve the safety of animal products by establishing a framework of risk management, throughout the food chain, which will reduce risks by eliminating or controlling hazards during the first stages of processing. The activities of Veterinary Services, from the organisational, technical, financial and human resource points of view, must be guided by quality assurance considerations, pursuant to the standards laid down in the Terrestrial Animal Health Code of the OIE. The quality of Veterinary Services depends therefore on a range of factors, including fundamental ethical, organisational and technical principles. It goes without saying that ethical principles cannot be effective unless the Veterinary Services have the resources necessary to reinforce them. The implementation of quality assurance by the Veterinary Services in developing countries, and particularly in Africa, will promote better organisation, effective use of limited resources, and professionalism in decision making. It will also allow for better access to international markets thanks to certification recognised by the OIE and the international community. PMID- 15884598 TI - Quality assurance applied to animal disease surveillance systems. AB - Monitoring and surveillance systems (MOSS) are essential activities for official Veterinary Services. In addition, the increased trade in animals and animal products over recent years has increased the importance of international disease reporting. A reliable surveillance system is the key to early warning of a change in the health status of any animal population. Such a system is also essential for providing evidence about the absence of diseases or in determining the extent of a disease which is known to be present. The authors discuss a set of methods and approaches for evaluating the quality of surveillance and survey systems. Certain steps are required when assessing the quality of a service or product. Various approaches for quality assessment are available and the suitability of each method depends on the objective of the evaluation. An essential basic requirement is, however, to use an objective, transparent and systematic approach. The evidence collected and the analyses used to reach conclusions must be of such high quality that the results are acceptable to both the management of the MOSS and the assessor. Repeated discussions and negotiations may be necessary to reach consensus, particularly if the judgement affects activities between trading partners. Well-documented MOSS with specified objectives and integrated quality assurance mechanisms are likely to be easier to evaluate. PMID- 15884599 TI - Quality assurance and meat inspection in Australia. AB - The aim of meat inspection in Australia is to contribute to the production of safe and wholesome food, comply with the requirements of importing countries, and support national animal health objectives. An analysis of the role of quality assurance (QA) in the meat inspection systems at federally inspected establishments shows that the position of Australia as a leading meat exporter is aided by a co-regulatory, QA-based approach to meat inspection, which is equally applicable to all species at slaughter. Technical developments in meat inspection at the national and international level during the 1990s led to significant enhancements in QA systems. Quality assurance is implemented through nationally uniform documented systems, which are designed to achieve consistent standards of meat safety. These systems are complemented by hazard analysis critical control point-based QA programmes which meet the quality-standards of the International Organization for Standardization. Quality assurance programmes aim for a 'whole of chain' approach, so that the system is implemented 'from the paddock to the plate', or from pre-harvest through to post-harvest, i.e., from on-farm practices to the refrigeration, storage and transportation stages. The QA elements of meat inspection employed in production systems in Australia have significantly contributed to the consistent achievement of meat safety objectives that are appropriate to contemporary risks. PMID- 15884600 TI - Evaluating the resources of Veterinary Services. AB - The assessment of human and material resource capacity and capability remains a critical component of the evaluation of Veterinary Services. Human resource evaluation examines the skills, qualifications and competencies of staff members and helps to determine the optimum number of staff needed to provide an efficient and effective service. Organisational arrangements, ongoing training and performance evaluation are important. Material resources include funds for operational activity, accommodation (including laboratory facilities), equipment, communication and transport. Evaluation is a complex process. The structure and service delivery arrangements of Veterinary Services must be taken into account. Evaluation can be based principally on input, infrastructure and quality systems or on outputs and outcomes. It may be based on a combined approach involving all of these components. PMID- 15884601 TI - The construction and evaluation of a questionnaire to identify quantitative criteria for evaluating national Veterinary Services. AB - This article presents the first part of a large-scale study aimed at developing a method for collecting standard information, based on quantitative criteria, about the national Veterinary Services of Member Countries of the OIE (World organisation for animal health). This information can then be used to assess such Veterinary Services in terms of quality and performance, or for accreditation. The study identified objective and relevant criteria for characterising Veterinary Services and an extensive questionnaire was developed, covering all aspects of the structures and functions of national Veterinary Services, which was sent to a broad range of OIE Member Countries. Each question and its answers were then evaluated and validated. In this way, 49 questions suitable for obtaining quantitative criteria to describe Veterinary Services were identified. In the next step of the study, this complex set of data will be further analysed. PMID- 15884602 TI - Tools for evaluating Veterinary Services: an external auditing model for the quality assurance process. AB - The author describes the reasons why evaluation processes should be applied to the Veterinary Services of Member Countries, either for trade in animals and animal products and by-products between two countries, or for establishing essential measures to improve the Veterinary Service concerned. The author also describes the basic elements involved in conducting an evaluation process, including the instruments for doing so. These basic elements centre on the following:--designing a model, or desirable image, against which a comparison can be made--establishing a list of processes to be analysed and defining the qualitative and quantitative mechanisms for this analysis--establishing a multidisciplinary evaluation team and developing a process for standardising the evaluation criteria. PMID- 15884603 TI - Approaches to resolving trade disputes. AB - The authors discuss the various approaches to resolving trade disputes available to Member Countries of the OIE (World organisation for animal health). The paper first describes the rights and obligations of Member Countries in setting health measures for the importation of animals and animal products, according to the provisions of the World Trade Organization (WTO) Agreement on the Application of Sanitary and Phytosanitary Measures (the SPS Agreement). The authors indicate how OIE standards may be used to set import measures and introduce issues such as equivalence and the use of provisional measures, which are both areas of potential conflict. The authors then describe the options available for resolving disputes--bilateral discussions, mediation through the OIE, the use of the WTO SPS Committee and the formal WTO dispute settlement process, discussing the advantages and disadvantages of each. PMID- 15884604 TI - Accreditation of veterinary inspection systems. AB - Accreditation of Veterinary Services could be a key factor in facilitating international recognition of certificates. For this to occur, accreditation must be conducted within the international normative framework on compliance evaluation. The EN 45004 standard for inspection bodies is the most appropriate organisational reference. It makes a clear distinction between the technical activity of inspection, and decisions which fall within the competence of public authorities. However, there are few international normative texts describing inspection methods, which is a major obstacle to the development of a widely recognised accreditation system. Organisations that accredit inspection bodies exist in many countries, but there are no agreements on multilateral recognition. This paper describes the accreditation cycle and outlines the possibilities for accrediting networks or individual sites. PMID- 15884605 TI - Prescription medications: effect on healthcare costs and dentistry. PMID- 15884606 TI - Dentistry: evolution and progress. PMID- 15884607 TI - Las Vegas Institute to expand globally. PMID- 15884608 TI - The hygiene department: how to turn a loser into a winner. PMID- 15884609 TI - Method and dosimetry for thermolysis and removal of biofilm in the periodontal pocket with near-infrared diode lasers: a case report. PMID- 15884610 TI - The role of glass ionomers in minimally invasive restorative dentin. PMID- 15884611 TI - Impression and model techniques for complex prosthodontics. PMID- 15884612 TI - The art of temporization: facial changes through aesthetic dentistry. PMID- 15884613 TI - Makeovers are ageless. AB - "Older patients" are often treated with indifference when it comes to how they look. The result may often be a patient not receiving the care that his or her goals would dictate. In the featured case, the patient had received dental hygiene care in a dental office for years. The fact that the patient's daughter had to take the initiative to seek care for a very motivated patient indicates a lack of understanding and communication in the patient's original dental office. The treatment plan that met the patient's goals was presented and accepted. It involved a full-mouth reconstruction, eliminating partial dentures, opening a deep bite, treating a diastema with orthodontics, and constructing a new smile. This makeover lent itself to the current trend of improving a smile, but it accomplished much more. The patient now has a firm foundation for better chewing, but perhaps more importantly, the patient, through CAMBRA, has the knowledge to control the disease process that caused the breakdown of her original dentition. PMID- 15884614 TI - Form and function in cosmetic dentistry. AB - The results of the case presented meet the aesthetic requirement of the patient as well as the biomechanical and functional requirements for long-term comfort, function, and stability. If not for the extra time spent with the patient uncovering the relationship between her occlusion and headaches and gaining her confidence by solving that problem first, we never would have had the opportunity to build a relationship that would allow her to proceed confidently with the cosmetic treatment that she described as the "best darn holiday present ever!" PMID- 15884615 TI - Anterior application of an indirect composite: a doctor/technician liaison's perspective. PMID- 15884616 TI - Prion diseases and infection control precautions. PMID- 15884617 TI - Initial communication: your lifeline to practice success. PMID- 15884618 TI - Computer backup clones, and stem cells. PMID- 15884619 TI - The evolution of endodontic progress: making excellence a reflection of simplicity. PMID- 15884620 TI - Managing severe canal curvatures and apical impediments: an endodontic case study. PMID- 15884621 TI - L-2-hydroxyglutaric aciduria: a report of 29 patients. AB - L-2-hydroxyglutaric aciduria (L2HGA) is a chronic slowly progressive neurodegenerative disease characterized mainly by psychomotor developmental delay and cerebellar dysfunction. We report the clinical, biochemical, and neuroimaging features of 29 patients from 22 families. The mean age at the time of diagnosis was 13.4 years (2.5-32 years). The mean follow-up period of patients was four years (1.5-16 years). The main clinical findings were mental retardation and cerebellar involvement with ataxic gait and intentional tremor. Additional findings were mental retardation, macrocephaly and seizures. Diagnosis was confirmed by increased urinary excretion of L-2-hydroxyglutaric acid in all patients and highly specific magnetic resonance imaging (MRI) pattern showing subcortical leukoencephalopathy with bilateral high signal intensity in dentate nuclei and putamens. During the follow-up period, all patients had a static encephalopathy course. The underlying metabolic defect and the possible role of L 2-hydroxyglutaric acid are studied in a subgroup of these families and under evaluation for publication. PMID- 15884622 TI - Maple syrup urine disease-treatment and outcome in patients of Turkish descent in Germany. AB - Maple syrup urine disease (MSUD) is a rare autosomal recessive disorder that causes acute and chronic brain dysfunction because of a neurotoxic effect of the accumulating branched chain amino acids (BCAA) and their corresponding keto acids. Aim of the treatment is a rapid reversal of the neonatal decompensation and a stable long-term metabolic control obtained by a carefully adjusted BCAA low diet. In optimally treated patients, an unimpaired neurological and intellectual outcome is possible. Ten patients of Turkish origin suffering from MSUD are presently treated in the Metabolic Unit of the University Hospital in Dusseldorf, Germany. All patients show mild intellectual deficits; neurological impairment is rare. This paper aims to define the feasible standard of therapy and the resulting intellectual and psychosocial outcome achievable in MSUD patients of Turkish origin under high standard conditions of medical care for inborn errors of metabolism. PMID- 15884623 TI - Does simultaneous gastric and esophageal pH monitoring increase the diagnosis of gastroesophageal reflux disease? AB - The duration of gastric acidity may affect the interpretation of esophageal pH monitoring study. The aim of this study was to increase the sensitivity of pH monitoring by simultaneous gastric and esophageal pH recording. Fifty-seven patients were enrolled in the study. After the first analysis, the recording periods in which gastric pH was >4 were excluded and after this exclusion parameters were recalculated. Of the 57 patients, 14 (24.6%) (mean age, 70+/-4.6 years) were diagnosed as having gastroesophageal reflux disease with the use of conventional method. After correction, gastroesophageal reflux disease diagnosis was made in 6 additional patients (mean age, 2.4+/-2.4 years) and total number of patients with reflux increased to 20 (35.1%) (p=0.031). The mean percentage time of gastric pH>4 was significantly greater in children younger than 2 years of age than that in those older than 2 years of age (50.6%+/-15.2% vs 33.7%+/-18.1%) (p=0.001). Exclusion of periods in which gastric pH>4 affected the results more obviously in patients younger than 2 years. Simultaneous gastric and esophageal pH monitoring is useful for the diagnosis of gastroesophageal reflux disease, particularly in children younger than 2 years of age. PMID- 15884624 TI - Cardiac troponin I: is it a marker to detect cardiotoxicity in children treated with doxorubicin? AB - Doxorubicin has been used in the the treatment of malignant tumors in children. Its use is limited by cardiotoxic effects beyond a cumulative dose of 450 mg/m2. To detect cardiotoxicity at an early stage and identify patients at risk for development of cardiotoxicity are matters of concern. Recently, cardiac troponin I (cTnI) has been reported to be useful for detecting minor myocardial damage. In the present study, we investigated whether cumulative doxorubicin-related myocardial cell damage can potentially increase cTnI levels above the expected values in 22 patients treated with cumulative doxorubicin doses of 120 to 450 mg/m2. Impaired cardiac functions were found in three patients by echocardiography, but serum CTnI levels were within the ranges expected in healthy individuals both in patients with cumulative doxorubicin doses > or = 400 mg/m2 and in patients with disturbed cardiac functions. We found no relationship between serum cTnI, cumulative dose of doxorubicin, and echocardiographical findings. PMID- 15884625 TI - Fatty liver in obese children: prevalence and correlation with anthropometric measurements and hyperlipidemia. AB - The aim of this study was to evaluate the correlation of ultrasonography-proven fatty liver with liver functions, serum lipid levels and anthropometric measurements in children with exogenous obesity. Three hundred and twenty-two patients (183 girls, 56.8%) with a mean age of 11.4+/-3.2 years (4-18 years) who presented with the complaint of obesity were enrolled. In 38 (11.8%) patients, increased liver echogenicity resembling fatty liver was found (Group 1). The body mass index percentages of group 1 patients were significantly higher than of those without fatty liver (Group 2) (157.7+/-18.0 vs 151.3+/-17.8, p=0.038). Alanine and aspartate aminotransferase levels of group 1 patients were significantly higher than of group 2 (p=0.002 vs p=0.028, respectively). Triglyceride levels were significantly higher in group 1 patients (120.8+/-88.8 vs 100.5+/-58.5 mg/dl, p=0.044). In conclusion, ultrasonography is an easy and noninvasive method for the diagnosis of fatty liver in children with obesity. Body mass index and serum lipids were higher in group 1 patients. The diagnosis and early treatment of obesity in childhood is important for the prevention and better treatment of related complications. Thus, ultrasonography should be a part of the early evaluation of obese children. PMID- 15884626 TI - The carriage of group B streptococci in Turkish pregnant women and its transmission rate in newborns and serotype distribution. AB - The prevalence of group B streptococci (GBS) colonization was studied in 500 pregnant women and their newborn infants by collecting vaginal and rectal swabs from mothers, and umbilical and throat swabs from their infants. Forty-six isolates of GBS were obtained from mothers' specimens and eight from neonates. Maternal and infant colonization rates were found to be 9.2% and 1.6%, respectively. Vertical transmission rate was 15.2%. Additionally, serotypes and antimicrobial susceptibility of 54 isolates of GBS were determined. Type Ia, II and III were common serotypes among GBS isolates from mothers and infants. When evaluating the factors that affect GBS carriage, age, socio-economic status and education level of pregnant women were important for carriage, while use of intrauterine device and parity were unrelated. No resistance to ampicillin, penicillin, ceftriaxone or vancomycin was found by disk diffusion method. A high level of resistance against tetracycline was noted (91%). Although invasive serotypes are predominant, the rarity of GBS disease in Turkish infants may be due to low rates of maternal carriage or to their possessing protective levels of GBS-specific IgG antibody in their sera. PMID- 15884627 TI - Usefulness of the myocardial performance index (MPI) for assessing ventricular function in obese pediatric patients. AB - Obesity is a prevalent pathological and nutritional disease relating to clinical and sub-clinical disorders. Furthermore, its complications exert a major impact on cardiovascular risk. Myocardial performance index (MPI) has been described as noninvasive Doppler measures of left ventricular (LV) function. The purpose of this study was to assess MPI using transthoracic Doppler echocardiography in normotensive obese and hypertensive obese pediatric patients and in a control group, and to investigate the relationship between MPI and LV mass, and LV systolic and diastolic functions. The study group consisted of 25 healthy cases (M/F: 13/12) as a control group (Group I), 25 normotensive-obese patients (M/F: 13/12) (Group II) and 25 hypertensive-obese patients (M/F: 14/11) (Group III). The mean ages were 12.1+/-1.8, 11.9+/-1.5 and 12.4+/-1.4 years, respectively. Cholesterol levels and body mass index (BMI) were higher in the hypertensive obese group. The MPI values were 0.37+/-0.04, 0.43+/-0.08 and 0.51+/-0.11 in the three groups, respectively. MPI measurements, LV mass fractional shortening (FS), ejection fraction (EF) and mitral E/A ratio were found significantly different, especially in the hypertensive-obese group. These findings may be important to determine the relationship between obesity and cardiovascular risk factors in pediatric ages. MPI may be useful in determining the relationship between them. PMID- 15884628 TI - The relationship between urinary calcium, sodium, and potassium excretion in full term healthy newborns. AB - The objective of this study was to determine the specific reference values for urinary calcium/creatinine (UCA/Cr) (mg/mg) in healthy breast-fed newborns, and to evaluate the relationship between UCa/Cr, urinary sodium/creatinine (UNa/Cr), urinary potassium/creatinine (UK/Cr) and UNa/UK ratios in the same group. A total of 88 infants aged between 0-28 days were enrolled in this study. They were divided into two age groups as follows: Group I: < or = 7 days of age; Group 2 infants aged between 8-28 days. Non-fasting spot urine was analyzed for Ca, Na, K and Cr. Significant differences were observed between the two groups in terms of UCa/Cr (0.11+/-0.10 vs 0.27+/-0.23, p<0.001), UNa/Cr (1.29+/-1.63 vs 5.5+/-4.83, p<0.001), and UK/Cr (0.94+/-0.99 vs 2.82+/-2.3, p<0.001). The data showed positive correlation between UCa/Cr and age (r=0.38, p<0.001) as well as between age and UNa/Cr ratio (r=0.68, p=0.0001) and between age and UK/Cr ratio (r=0.57, p<0.0001). Additionally, there was a positive correlation between UNa/UK and age (r=0.40, p=0.001). The UCa/Cr ratio positively correlated with UNa/Cr whereas no correlation was found between UCa/Cr and UNa/Uk ratio. Our data suggest that the healthy neonates differ from the hypercalciuric patients by exhibiting a linear correlation between Na/K and UCa/Cr. As the normal values of UCa/Cr, UNa/Cr, UK/Cr, UNa/UK ratios in the early neonatal period differ from those in the late neonatal period, these differences should be taken into consideration when assessing urinary excretion of these parameters for diagnostic purposes in the early and late newborn periods. PMID- 15884629 TI - Mothers' attitudes towards their own presence during invasive procedures on their children. AB - This study assessed of the preferences 742 mothers regarding their own presence during invasive procedures performed on their children. The relationships between socio-demographical characteristics and preferences of the mothers and disease characteristics of the children were examined. A mother's desire to be present was found to increase with decreasing invasiveness of the procedure as well as with increasing analgesia and sedation provided. The desire to be present was higher in young mothers with higher socio-economic levels and educational backgrounds, with younger children and with children who had undergone prior recurrent interventions. This study demonstrated that most of the mothers preferred to be present during the procedure, and that the ratio of mothers willing to do so increased significantly if the children were sedated. The results suggested that pediatricians can improve the quality of service and physician-patient-family relationship by taking mothers' preferences into consideration. PMID- 15884630 TI - Encopresis: long-term clinical outcome of 67 cases. AB - In this study we attempted to investigate the outcome of encopresis and to determine factors affecting prognosis. The sample consisted of 52 boys (77.6%) and 15 girls (22.4%) diagnosed as encopresis according to DSM IV diagnostic criteria. These patients were evaluated six years after their initial examination in the Department of Child Psychiatry. Clinical and demographical data were compared between initial and follow-up interviews and between patients with complete recovery and others. Fifty-six patients (83.6%) recovered completely and 11 (16.4%) continued to be encopretic after six years. Good school performance (p<0.005), high levels of parental education (p<0.005) and absence of constipation (p<0.05) were associated with favorable outcome. In addition, secondary encopretics who were diagnosed within a year from onset of the symptom recovered significantly earlier (p<0.001). Encopresis is a chronic disorder and complete recovery rates tend to increase with time. Families and primary health care providers should be informed about the treatment possibilities of encopresis for early intervention. PMID- 15884631 TI - Subacute sclerosing panencephalitis (SSPE) associated with congenital measles infection. AB - A 13-month-old male presented with repetitive episodes of myoclonic jerks of the head and extremities for two months. His past medical history revealed that his non-immunized mother had measles at the time of delivery. Measles antibody titers in serum and cerebrospinal fluid (CSF) were 1/512 (hemagglutinin inhibition [HI]) and 1/128 HI, respectively. Immunofixation electrophoresis of CSF revealed an oligoclonal IgG band. The magnetic resonance imaging (MRI) of the brain on T2 weighted images showed lesions of high signal intensity in the subcortical white matter. Electroencephalography (EEG) revealed periodic high-amplitude slow waves. Diagnosis of subacute sclerosing panencephalitis (SSPE) was based upon clinical presentation, a characteristic EEG, and abnormal CSF studies. MRI findings supported the diagnosis. To the best of our knowledge, this is only the 3rd case to date, of SSPE-associated congenital measles in the literature. PMID- 15884632 TI - Neonatal lupus erythematosus presenting with cholestatic hepatitis: a case report and review of the literature. AB - Neonatal lupus erythematosus (NLE) is a disease primarily characterized by cardiac and/or cutaneous involvement. Hepatic, hematological, neurological and pulmonary involvement are rare manifestations and normally considered as mild and transient complications. But recent studies have shown more frequent hepatic involvement in NLE. We report a two month-old male infant, born to a clinically asymptomatic mother, presenting with significant hepatic involvement and annular, erythematous plaques with hyperkeratotic borders at the eyebrow region and anterior surface of trunk. Both the infant and his mother were positive for anti Ro (SS-A) and anti-La (SS-B). PMID- 15884633 TI - Clinical and pathological aspects of ARC (arthrogryposis, renal dysfunction and cholestasis) syndrome in two siblings. AB - We describe the first family report of ARC syndrome (arthrogryposis multiplex congenita, renal dysfunction, and cholestasis) diagnosed in Turkey. ARC syndrome is a rare cause of cholestatic jaundice and skeletal abnormalities in the neonatal period. Fanconi-like renal tubular dysfunction completed the clinical picture. Consanguinity and affected membership are the other typical components of this rare disorder, and possibility of autosomal recessive transmission was considered. A broad spectrum of histopathological abnormalities have been described in the liver and kidney. In this report, we describe two male siblings with ARC syndrome who had cholestatic jaundice, arthrogryposis multiplex congenital-like joint contractures and renal involvement with additional clinical features. Clinical and pathological aspects of the syndrome are discussed and compared with the other cases in the literature. PMID- 15884634 TI - Cutaneous intravascular papillary endothelial hyperplasia of the hand: a case report and review of the literature. AB - Intravascular papillary endothelial hyperplasia (IPEH) is a rare benign endothelial proliferation that bears a remarkable resemblance to angiosarcoma. We report a case of cutaneous IPEH in the thumb of a seven-year old girl which was successfully treated with excision. The relevant literature is reviewed and histopathological features, differential diagnosis and natural history of the lesion are discussed. PMID- 15884635 TI - Intralesional corticosteroid injection for central giant cell granuloma: a case report and review of the literature. AB - Central giant cell granuloma (CGCG) is a benign intraosseous lesion of the jaws that is found predominantly in children and young adults. Although benign, it may be locally aggressive, causing extensive bone destruction, tooth displacement and root resorption. The common therapy is aggressive curettage, peripheral ostectomy or resection, which may be associated with loss of teeth and, in younger patients, loss of dental germs. A number of alternative nonsurgical approaches have been advocated in recent years for the management of CGCGs. These include intralesional corticosteroid injections, calcitonin injections and subcutaneous alpha-interferon injections. In this article, an 1-year-old boy with a CGCG is successfully treated with corticosteroid injections and this treatment is discussed within a review of the literature. PMID- 15884636 TI - Congenital aneurysm of the muscular interventricular septum in a child. AB - Congenital aneurysms of the muscular interventricular septum without ventricularseptal defect are extremely rare. We describe a five-month-old girl patient with congenital aneurysm, pathological ST changes and minimal right ventricular outflow tract obstruction. The electrocardiogram showed Wolf Parkinson-White syndrome, pathological ST changes and combined ventricular hypertrophy criteria. The transthoracic echocardiography and cardiac catheterization demonstrated a septal aneurysm which was bulging into the right ventricle and systolic pressure gradients between the main pulmonary artery and right ventricle outflow tract with normal coronary arteries. The patient has shown no symptoms, but some rhythm disturbances may be expected because of her electrocardiographic anomalies. Therefore, these patients should be followed carefully because of possible complications. PMID- 15884637 TI - A case with ICF syndrome lost to rubella pneumonitis. AB - The immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome is a rare autosomal recessive disorder characterized by variable immunodeficiency, instability of the pericentromeric heterochromatin, and facial dysmorphism. Here we report a new case of ICF syndrome who died of rubella pneumonitis. A six year-old-girl who was the first child of consanguineous parents was admitted to the hospital because of bronchopneumonia. Laboratory investigations revealed pan-hypogammaglobulinemia, lymphoperria, normal proportions of peripheral blood lymphocytes with an inverted CD4/CD8 ratio, and interstitial pneumonia with a positive serology of acute rubella infection. The ICF syndrome was diagnosed by centromeric instability in the standard cytogenetic analysis. An inclusion body was demonstrated in the lung biopsy after the death of the patient. Chromosomal investigation could be helpful along with other tests for diagnosis of variable immunodeficiency accompanied by facial dysmorphism. PMID- 15884638 TI - Raine syndrome associated with cytomegalovirus infection. AB - Intracranial calcification, periosteal proliferation and microcephaly, which represent the clinical features of the congenital cytomegalovirus infection, can also be seen in a rare disorder named Raine syndrome. This clinical entity has been reported in eight families so far. Here, we report a new patient with clinical features of the Raine syndrome associated with cytomegalovirus infection. Although this may be a chance association only, the condition raised the question of whether early intrauterine CMV infection may contribute to the Raine phenotype. PMID- 15884639 TI - Magnetic resonance angiographic and three-dimensional computerized tomographic identification of scimitar syndrome in an 8-month-old infant. AB - Scimitar syndrome is a rare congenital abnormality. Clear anatomic definition is an important issue before operation. We report definition of both abnormal pulmonary venous drainage and anomalous systemic arterial supply by gadolinium enhanced magnetic resonance angiography and three-dimensional computerized tomography in an eight month-old infant with scimitar syndrome. The presented case study confirms that these noninvasive diagnostic tools can successfully be used to provide valuable information about vascular anatomy in infants with scimitar syndrome. PMID- 15884640 TI - A new case of omphalocele with absence of thumb. AB - Omphalocele, or exomphalos, is a rare defect of the periumbilical abdominal wall in which the coelomic cavity is covered with peritoneum only, resulting in congenital eventration of abdominal contents at the base of the umbilical cord. The etiology is still unknown. In this paper a new case of omphalocele with absence of the right thumb in a five-year-old boy is reported. A common etiology and a new terminology for this combination are suggested. PMID- 15884641 TI - Secondary glioblastoma multiforme with a new translocation t(3;3)(q21;q26) following treatment of acute lymphoblastic leukemia. PMID- 15884642 TI - A case of tuberculous peritonitis with elevated CA 125. PMID- 15884643 TI - CVMA's international role. PMID- 15884644 TI - An ethicist's commentary on the organic dairy farmer who will not use antibiotics for foot rot. PMID- 15884646 TI - Bilateral flexor tendon contracture following onychectomy in 2 cats. AB - Two cats presented with bilateral flexor tendon contracture following onychectomy. This previously unreported complication proved to be painful and debilitating. Deep digital flexor tenectomy successfully resolved the problem. Twelve months after surgery, the first cat remains free of complications. The second cat recovered full limb function, but died of unrelated causes. PMID- 15884645 TI - A review of Neospora caninum in dairy and beef cattle--a Canadian perspective. AB - Neospora caninum is one of the most important causes of abortion in cows. The occurrence of N. caninum infection in beef and dairy cattle has been reported worldwide, and in most provinces in Canada. The objective of this review is to summarize our current understanding of N. caninum in dairy and beef cattle for Canadian bovine practitioners. The review covers the life cycle of the agent, its mechanisms of transmission, clinical signs, and tests for diagnosing the infection. Data on the prevalence of the infection in Canadian dairy and beef cattle are reviewed and briefly compared with estimates from other parts of the world. Most importantly for Canadian bovine practitioners, the impacts of the infection, risk factors for its occurrence, and methods of control are also discussed. By reviewing the scientific literature on N. caninum from a Canadian perspective, culling decisions based on the interpretation of diagnostic tests are more effectively made in the control of N. caninum-associated disease. PMID- 15884647 TI - Bladder outlet obstruction in a 6-month-old alpaca secondary to pelvic displacement of the urinary bladder. AB - A 6-month-old female alpaca was presented for stranguria. Based on the history, physical examination findings, and radiographic studies, the alpaca was diagnosed with bladder outlet obstruction, secondary to pelvic displacement of the bladder, a condition previously unreported in camelids. Cystopexy was performed and the alpaca recovered unremarkably. PMID- 15884648 TI - A preliminary equine abuse policy with potential application to veterinary practice. AB - An equine abuse policy was developed as an adjunct to an equine management survey. If at least 3 of 5 categories caused concern, a report to the authorities was indicated. The policy was not used but, in the absence of other guidelines, it might assist veterinarians considering potential abuse cases. PMID- 15884649 TI - A comparison of the cytobrush and uterine lavage techniques to evaluate endometrial cytology in clinically normal postpartum dairy cows. AB - This study compares cytobrush and lavage techniques for the assessment of endometrial cytology (EC) in clinically normal postpartum dairy cows. The EC samples were collected from Holstein cows (n = 35) during visit 1 (V1) at 20 to 33 d in milk (DIM) and 2 wk later during visit 2 (V2) at 34 to 47 DIM by using both techniques. A minimum of 100 cells were counted to determine the percentage of cells that were neutrophils (%PMN). The mean %PMN was significantly different between the techniques at V1 (P = 0.001), but not at V2 (P = 0.474). Overall, the %PM N decreased with time postpartum (r2 = 0.36; P = 0.001), but not within V1 (P > 0.05) or V2 (P > 0.1). Uterine diameter was negatively correlated with fluid recovery by the lavage technique (r2 = 0.41; P = 0.002). The mean %PMN was not influenced by the volume of fluid recovered in successful attempts, but 17% (12/70) of attempts yielded no fluid. In conclusion, the cytobrush technique is a consistent and reliable method for obtaining endometrial samples for cystologic examination from postpartum dairy cows. PMID- 15884650 TI - Bile peritonitis associated with gastric dilation-volvulus in a dog. AB - This report describes a case of septic peritonitis and gall bladder rupture in German shepherd dog that developed 7 d after surgical treatment for gastric dilation-volvulus. Histological examination confirmed gall bladder necrosis, secondary to an acute ischemic event. Postoperative acute necrotizing cholecystitis has not been a previously reported complication in dogs. PMID- 15884651 TI - Severe lymphocytic-plasmacytic and atrophic gastritis, as well as, predominantly eosinophilic, severe enteritis, in a 19-month-old Labrador retriever. AB - A 19-month-old, male, Labrador retriever was presented with a history of chronic vomiting. Blood analysis and abdominal ultrasononography could not rule out the possibility of gastrointestinal inflammation. Histologic examination of endoscopic gastrointestinal biopsies confirmed severe lymphocytic-plasmacytic and atrophic gastritis, as well as, predominantly eosinophilic, severe enteritis. PMID- 15884652 TI - The valuation of a veterinary practice. PMID- 15884653 TI - Magnetic resonance imaging of GM2-gangliosidosis in a golden retriever. PMID- 15884654 TI - Limbal melanocytoma of the left eye. PMID- 15884655 TI - Responses to chemical, physical and biological stresses: involvement of extracellular alarmones, pheromones and varisensors. PMID- 15884656 TI - Ribosome inactivation for preservation: concepts and reservations. AB - The role of the bacterial ribosome in the cellular response to environmental stress has been widely considered over last decade. Certain ribosome-associated proteins have been shown to induce conformational changes that lead to the formation of inactive forms of ribosomes that are presumed to be more stable during stationary phase. This was found to aid the survival of bacteria in this phase. Such proteins include ribosome modulation factor (RMF), YfiA and YhbH. Examining the influence of RMF on the survival of E. coli under heat, acid and osmotic stress showed that it was important for bacterial viability under these environmental pressures. However, the mechanism by which this protein exerts its effect has not been fully elucidated. The present work reviews the involvement of ribosomes in determining cell behaviour during stress. It focuses on the action of the ribosome-associated proteins and their role in inactivating ribosomes for preserving their integrity and aiding cell survival under stress. PMID- 15884657 TI - Applied and ecological aspects of oxidative-stress damage to bacterial spores and to oral microbes. AB - Bacterial cells have adapted in a variety of ways to resist oxidative stresses and damage in their everyday lives in a predominantly aerobic world. The nearly universal occurrence of resistance mechanisms against oxidative stresses, particularly those due to reactive oxygen species (ROS), suggests that most, if not all, bacteria have to deal with oxidative assaults. A primary source of oxidative stress is aerobic metabolism, which leads to production of ROS such as hydrogen peroxide, superoxide radical, perhydroxyl radical, hydroxyl radical and a variety of other toxic metabolites, including organic peroxides and other organics or inorganics able to transfer electrons to sites of oxidative damage. Anaerobes as well as aerobic and facultative organisms are subject to oxidative stresses, often as a result of their own metabolism of O2 or that of associated facultative organisms. If anaerobes would just ignore oxygen instead of metabolizing it, they would not have to deal with toxic metabolites of their own making. Another major source of oxidative stress comes from the use of oxidative agents in the disinfection-sterilization industry. Notable examples are hypochlorite for water purification and hydrogen peroxide used for industrial sterilization. Antimicrobials such as isoniazide and mitomycin C also act oxidatively to cause damage. In this article, aseptic packaging and processing involving use of hydrogen peroxide for sterilization of packaging materials is reviewed as an example of oxidative stress imposed on bacterial spores and vegetative cells from outside the organisms or the microbial community. The other example considered is related to oral microbiology and infectious disease in which oxidative stress may arise from the metabolism of the oral microbiota or may come from outside through use of oral care products. PMID- 15884658 TI - Bacterial sensitivity to bacteriophage in the aquatic environment. AB - There are several unusual features about phage when you first encounter them as a biologist. They are small, but conform to one of a few morphological types. Next their genomes can be composed of DNA or RNA and be single or double stranded. Finally they are numerically more abundant than prokaryotes and a significant proportion of them form an association in their microbial host populations termed lysogeny. The latter findings indicate that they are numerically significant in microbial populations. Since bacterial and phage abundance or lack of it is related in environments, this implies that the phage populations 'titrate' their hosts, and more probably the host's physiological status. Microbial populations wax and wane with nutritional inputs and there is a dynamic relationship between phage population sizes and host numbers and physiology. Overlay this with the different phage life cycle strategies, exemplified at the extremes by phage lambda (temperate) and phage T4 (virulent), then it becomes apparent that phage are a component in nutrient cycling in ecology. But their contribution does not stop there. Many are capable of transduction, moving DNA from one cell into another. So they can also aid the evolutionary progress of microbial populations by allowing them to share genes, just as gene exchange via plasmids and transformation does. Our perception of bacteria has been derived from pure culture studies and we are just being able to appreciate how subtle their ecological interactions are. This is no less true of the studies on bacteriophage, which are almost all based on laboratory experimentation, where the hosts are physiologically stressed by growing in 'high nutritional and optimum conditions'. The natural environment is naturally discontinuous and life evolved in this. Thus our perceptions of bacteriophage and their life cycle patterns derived from laboratory experimentation may be a little off the mark when we come to understand how they and their hosts interact in the niches available to them. It is worth just considering this as you read the article, as I suspect phage behaviours are more intimately involved in, and moderated by the physiological stresses in the life cycle of bacteria than we currently believe. PMID- 15884659 TI - Enterobacterial responses to external protons, including responses that involve early warning against stress and the functioning of extracellular pheromones, alarmones and varisensors. AB - Several striking findings, related to biological effects of external acidity, are reviewed here. The first of these relates to the role of PhoE in the penetration of H+ and protonated metabolites into the cell. PhoE is an anion pore and would not be expected to take up protons. The work reviewed here, however, shows that the loss or repression of PhoE leads to poor H+ passage through the outer membrane (OM), whilst derepression of PhoE leads to facilitated passage. It is now believed that H+ crosses through the PhoE pore in association possibly with oligopeptides, and that other protonated molecules, such as the acid tolerance EIC, use the same means to cross the OM. Additionally, several processes that form early warning systems against acidity are reviewed here. First, the properties of the acid tolerance EIC alarmones allow them to diffuse to regions not yet facing acid stress, and there give early warning and induce sensitive organisms to tolerance. Second, some agents, such as glucose, induce acid tolerance in organisms, long before these organisms are exposed to catabolically produced acidity, preparing them, in advance, to resist this impending acid challenge. Third, the occurrence of multiple forms of ESCs (i.e. of varisensors) ensures that where organisms have been grown under conditions that sensitise them to acid stress, the ESCs formed are modified so as to be activated at much higher pH values, ensuring that lethality by acid is reduced or abolished. Fourthly, normally only EICs induce tolerance. Strikingly, however, pH 8.5 or 9.0-grown cells are induced to tolerance by ESC formed at pH 6.5. This is believed to provide another early warning system, protecting alkali-grown cells against sudden acidification of media. Two other finding reviewed here should be emphasised. First, the hydrophobic antibiotic novobiocin is ineffective against enterobacteria, due to its failure to penetrate the OM barrier. This only applies to cultures in pH 7.0 media, however, cells growing at pH 5.0 being exquisitely sensitive to novobiocin, due to a conformational change to the antibiotic at acidic pH, which allows ready penetration through the OM. Second, acidic pHs affect the synthesis and effects of another antibiotic, namely colicin V. Thus pH 5.0 prevents both synthesis of this agent and its effects on sensitive cells. Exposure to external acidity leads to numerous other effects, including those that influence growth, cell division, plasmid transfer and chemotaxis; these have also been reviewed here. PMID- 15884660 TI - Progression to renal failure. AB - Chronic renal failure is characterized by a persistently abnormal glomerular filtration rate. It represents a developing process that is initiated by various causes, all with the common end result of persistent and usually progressive damage of varying severity to the kidneys. Although the causes and the locations of renal insult may vary, many features are common in progression to renal failure, and the final histological picture is one of glomerulosclerosis, interstitial fibrosis and loss of renal cells, mainly by apoptosis. The rate of progression varies significantly. However, individual genetic variations and disease-specific mechanisms contribute to the variability in rates of progression and response to therapy. Possible mechanisms of progressive renal damage include hemodynamic factors, hypertension, proteinuria, angiotensin II, and other chemical mediators such as cytokines and growth factors. Processes and predictive factors of progression to renal failure are discussed here along with clinical practice guidelines: Evaluation, Classification and Stratification. Prevention of the progression is also briefly mentioned. PMID- 15884661 TI - Growth and nutrition of children with chronic renal failure. AB - It has long been recognized that chronic renal failure (CRF) in children is associated with growth delay. Still In our days nevertheless, growth retardation remains today a major impediment to the full rehabilitation of children with CRF. The reduction of in height velocity frequently results in diminished final adult height. Available evidence suggests that growth retardation might be the result of late referral and/or suboptimal clinical care in children with CRF. Management of malnutrition, renal osteodystrophy, metabolic acidosis, salt wasting and anemia should be optimal before recombinant human growth hormone initiation. PMID- 15884662 TI - Management of renal osteodystrophy in children. AB - Prevention and treatment of renal osteodystrophy (ROD) are great challenges for pediatric nephrologists. The strategies for prevention and treatment of ROD in children with chronic renal failure (CRF) should be created on an individual basis. The following factors should be considered: age, type of primary disease, rate of progression of CRF, nutrition, acidosis, type of dialysis, and drugs (corticosteroids, growth hormone, etc). The treatment should start very early in the course of renal insufficiency with close monitoring of serum calcium, phosphate, alkaline phosphatase and parathormone (PTH) levels. Maintenance of serum phosphate within age- appropriate limits is essential for prevention of secondary hyperparathyroidism. PTH levels should be kept within normal limits in predialysis children and 2-3 times over upper normal limit in those on dialysis. Aggressive treatment with calcium-based phosphate binders and vitamin D derivates should be avoided to prevent PTH oversuppression and development of adynamic bone disease. The advantage in this respect is the development of calcium- and aluminum-free phosphate binders, of which there is limited pediatric experience with sevelamer hydrochloride. Paricalcitol is a non-hypercalcemic vitamin D analogue, and preliminary favorable experience has been reported in children. Calcimimetics like cinacalcet hydrochloride, which directly stimulate calcium sensing receptor and potently suppress PTH secretion without increasing plasma calcium in adults, are very promising agents, but pediatric experience is lacking. PMID- 15884663 TI - Management of renal anemia. AB - Normochromic normocytic anemia is common in children with chronic renal failure (CRF) when their glomerular filtration rate is below 35 ml/min/1.73 m2 BSA, but it may develop earlier in some forms of renal disease. An inadequate erythropoiesis due to insufficient erythropoietin synthesis in the kidneys is the main cause of renal anemia. Other reasons include reduced red blood cell lifespan, chronic blood loss, iron deficiency, inhibitors of erythropoiesis, and malnutrition. The presence of anemia contributes to many of the symptoms of uremia, including decreased appetite, decreased energy, poor cardiac function, and poor school performance. Therefore, correction of anemia dramatically improves the life of the child with CRF. Presently, the goal of anemia management is to maintain hematocrit concentrations at 33% to 36% and a hemoglobin concentration of at least 11 g/L. This can be accomplished by intravenous or subcutaneous administration of recombinant erythropoietin (rHuEPO, 100-300 U/kg/week) and iron preparations. If adequate iron stores cannot be maintained with oral therapy (2-3, max 6 mg/kg/day), intravenous iron should be administered. In order to optimize anemia management in children with CRF, future research should be concentrated on the normalization of hemoglobin early in the course of CRF, and the long-term effects on the child's development. PMID- 15884664 TI - Hypertension in children with chronic renal failure. AB - Systemic hypertension (HTN) is one of the major problems associated with chronic renal failure (CRF). HTN is a symptom and complication of CRF. The prevalence of HTN varies with the cause of CRF. The incidence of HTN increased up to 90% with progressive deterioration of the glomerular filtration rate (GFR). HTN is the major risk factor for decline in renal function and progression of CRF. It is the most important risk factor for cardiovascular diseases and morbidity and mortality in patients with CRF and end-stage renal disease (ESRD) on dialysis. The target blood pressure for hypertensive children with CRF should be under the 95th percentile for sex and age. The therapeutic approach in CRF is directed at reducing volume expansion and sodium retention, and decreasing peripheral vascular resistance. Diuretics are first-line therapy for HTN in patients with CRF with sodium and water retention. ACE inhibitors are the first-class drugs because of their renoprotective effect in preventing deterioration of kidney function. Calcium channel blockers are excellent first-line antihypertensive drugs. Recently angiotensin II receptor blockers and ACE inhibitors have been efficiently used together for the treatment of HTN and to prevent further decline in renal function. PMID- 15884665 TI - Impossible problems. PMID- 15884666 TI - Tandem mass spectrometry for peptide and protein sequence analysis. PMID- 15884667 TI - Immunodetection of 5'-methylcytosine on Giemsa-stained chromosomes. PMID- 15884668 TI - Shuttle system allowing simplified cloning of expression cassettes into advanced generation lentiviral vectors. PMID- 15884669 TI - URSA: ultra rapid selection of antibodies from an antibody phage display library. PMID- 15884670 TI - UBViz: a software tool for exploring metabolic pathways in 3-D space. PMID- 15884671 TI - SGCaller: a program to call and review genotypes measured by sequencing. PMID- 15884672 TI - NANEV: a program employing evolutionary methods for the design of nucleic acid nanostructures. PMID- 15884673 TI - Single-molecule dilution and multiple displacement amplification for molecular haplotyping. AB - Separate haploid analysis is frequently required for heterozygous genotyping to resolve phase ambiguity or confirm allelic sequence. We demonstrate a technique of single-molecule dilution followed by multiple strand displacement amplification to haplotype polymorphic alleles. Dilution of DNA to haploid equivalency, or a single molecule, is a simple method for separating di-allelic DNA. Strand displacement amplification is a robust method for non-specific DNA expansion that employs random hexamers and phage polymerase Phi29 for double stranded DNA displacement and primer extension, resulting in high processivity and exceptional product length. Single-molecule dilution was followed by strand displacement amplification to expand separated alleles to microgram quantities of DNA for more efficient haplotype analysis of heterozygous genes. PMID- 15884674 TI - Cell line fingerprinting using retroelement insertion polymorphism. AB - Human cell lines are an indispensable tool for functional studies of living entities in their numerous manifestations starting with integral complex systems such as signal pathways and networks, regulation of gene ensembles, epigenetic factors, and finishing with pathological changes and impact of artificially introduced elements, such as various transgenes, on the behavior of the cell. Therefore, it is highly desirable to have reliable cell line identification techniques to make sure that the cell lines to be used in experiments are exactly what is expected. To this end, we developed a set of informative markers based on insertion polymorphism of human retroelements (REs). The set includes 47 pairs of PCR primers corresponding to introns of the human genes with dimorphic LINE1 (L1) and Alu insertions. Using locus-specific PCR assays, we have genotyped 10 human cell lines of various origins. For each of these cell lines, characteristic fingerprints were obtained. An estimated probability that two different cell lines possess the same marker genotype is about 10-18. Therefore, the proposed set of markers provides a reliable tool for cell line identification. PMID- 15884675 TI - Minimizing DNA contamination by using UNG-coupled quantitative real-time PCR on degraded DNA samples: application to ancient DNA studies. AB - PCR analyses of ancient and degraded DNA suffer from their extreme sensitivity to contamination by modern DNA originating, in particular, from carryover contamination with previously amplified or cloned material. Any strategy for limiting carryover contamination would also have to be compatible with the particular requirements of ancient DNA analyses. These include the need (i) to amplify short PCR products due to template fragmentation; (ii) to clone PCR products in order to track possible base misincorporation resulting from damaged templates; and (iii) to avoid incomplete decontamination causing artifactual sequence transformation. Here we show that the enzymatic decontamination procedures based upon dUTP- and uracil-N-glycosylase (UNG) can be adapted to meet the specific requirements of ancient DNA research. Thus, efficiency can be improved to vastly reduce the amplification of fragments < or = 100 bp. Secondly, the use of an Escherichia coli strain deficient in both UNG and dUTPase allows for the cloning of uracil-containing PCR products and offers protection from plasmid DNA contamination, and, lastly, PCR products amplified from UNG-degraded material are free of misleading sequence modifications. PMID- 15884676 TI - Normalization of soil DNA extraction for accurate quantification of target genes by real-time PCR and DGGE. AB - The analysis of microbial communities in environmental samples requires accurate and reproducible methods for extraction of DNA from sample matrices that have different physical and chemical characteristics. Even with the same sample type, variations in laboratory methods can result in different DNA yields. To circumvent this problem, we have developed an easy and inexpensive way to normalize the quantities of DNA that involves the addition of an internal standard prepared from plasmid DNA. The method was evaluated by comparing DNA yields using different DNA extraction procedures, after which the DNA was used for microbial community analysis by PCR-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S ribosomal RNA (rRNA) and for quantification of 16S rRNA gene copy numbers in environmental samples by real-time PCR. Our results show that use of the internal standard allows normalization of the resulting data and more accurate quantification of gene copy numbers in soil samples. These methods should also have broad application for various other types of environmental samples. PMID- 15884677 TI - Taking arrays from the lab to the field: trying to make sense of the unknown. AB - With the rapid pace of nucleic acid microarray technology development and a renewed national emphasis on detecting and characterizing microorganisms in environmental samples, there is a rush to operationalize existing microarray technologies and apply them to uncharacterized environmental backgrounds. The purpose of this article is to pause and ask a basic question: what do microarray data actually mean in the face of uncharacterized sample backgrounds? In attempting to answer this question, we draw a clear distinction between hypothesis-driven fundamental science and operational uses of microarray technology; assess microarray technology assumptions in the face of uncharacterized environments; offer an environmental microbiologist's perspective on technology needs and requirements for quantitatively analyzing microbial communities; and hopefully stimulate a scientific and technical dialogue around the concept of analytical environmental microbiology and future technology development. PMID- 15884678 TI - Locked nucleic acid inhibits amplification of contaminating DNA in real-time PCR. AB - Locked nucleic acid (LNA) is a modified DNA with increased binding affinityfor complementary DNA sequences. Our strategy was to use this property of LNA to inhibit undesired PCR amplification (e.g.,from contaminating genomic DNA) in a cDNA-based assay. By placing a short complementary LNA sequence in intronic DNA, the aim was to inhibit the amplification of genomic DNA without affecting the amplification of reverse-transcribed spliced mRNA. LNA was designed to bind within intron 5 in the x-box binding protein 1 (XBP1) gene. An irrelevant LNA oligonucleotide served as a negative control. In both PCR and real-time PCR, the addition of LNA showed blocking of the amplification of genomic XBP1 but not cDNA XBP1. To test the effect of melting temperature (Tm) on the LNA, we investigated the number of LNA nucleotides that could be replaced with DNA nucleotides and still retain the blocking activity. More than three DNA nucleotides reduced the LNA inhibition ability. The sequence specificity of the LNA was tested by investigating the number of LNA nucleotide mismatches permitted. The introduction of one mismatch maintained the inhibition of genomic amplification whereas two mismatches reduced the amplification. Our results show that LNA may be used to enhance the specificity of PCR by eliminating unwanted PCR products. PMID- 15884679 TI - VECT: an automatic visual Perl programming tool for nonprogrammers. AB - Modern high-throughput biological research produces enormous amount of data that must be processed by computers, but many biologists dealing with these data are not professional programmers. Despite increased awareness of interdisciplinary training in bioinformatics, many biologists still find it difficult to create their own computational solutions. VECT, the Visual Extraction and Conversion Tool, has been developed to assist nonprogrammers to create simple bioinformatics without having to master a programming language. VECT provides a unified graphical user interface for data extraction, data conversion, output composition, and Perl code generation. Programming using VECT is achieved by visually performing the desired data extraction, conversion, and output composition tasks using some sample user data. These tasks are then compiled by VECT into an executable Perl program, which can be saved for later use and can carry out the same computation independently of VECT. VECT is released under the GNU General Public License and is freely available for all major computing platforms including Macintosh OS X, Linux, and Microsoft Windows at www.complex.iastate.edu. PMID- 15884680 TI - Adenovirus-based short hairpin RNA vectors containing an EGFP marker and mouse U6, human H1, or human U6 promoter. PMID- 15884681 TI - Alternative approach to generate shRNA from cDNA. AB - Short hairpin RNA (shRNA) synthesized from vector-based expression is as effective as short interfering RNA (siRNA) synthesized in vitro for suppressing the expression of their corresponding genes. Recently, three groups independently reported a new technology to construct an shRNA library from cDNA, providing great hope for genome-wide functional screens in many biological systems. In the present study, we report an alternative approach to generate shRNA from cDNA. A major improvement was to use a nicking enzyme to open up the double-stranded DNA so that the loop region remains single-stranded while the rest of the DNA fragment is double-stranded at an elevated temperature (e.g., 72 degrees C). The single-stranded DNA was then converted into double-stranded DNA by Taq DNA polymerase using the existing strand in the double-stranded region as a primer. Thus, the extended product carried a palindromic structure of 19 bp separated by a loop. Finally, the DNA fragment was cloned into a vector that carries an H1 promoter at the upstream region and ends with 5Ts, a terminator for the Pol III polymerase, at the downstream region. To prove the principle, we constructed shRNA from green fluorescent protein (GFP) cDNA and successfully suppressed GFP expression. Consequently, this simplified approach provides a better alternative to generate shRNA libraries from cDNA. Such shRNA libraries can be used to identify potential siRNA target sequences and study gene functions by a variety of selection methods. PMID- 15884682 TI - Colorimetric approach to high-throughput mutation analysis. AB - High-throughput genomic mutation screening for primary tumors has characteristically been expensive, labor-intensive, and inadequate to detect low levels of mutation in a background of wild-type signal. We present a new, combined PCR and colorimetric approach that is inexpensive, simple, and can detect the presence of 1% mutation in a background of wild-type. We compared manual dideoxy sequencing of p53 for eight lung cancer samples to a novel assay combining a primer extension step and an enzymatic colorimetric step in a 96-well plate with covalently attached oligonucleotide sequences. For every sample, we were able to detect the presence or absence of the specific mutation with a statistically significant difference between the sample optical density (OD) and the background OD, with a sensitivity and specificity of 100%. This assay is straightforward, accurate, inexpensive, and allows for rapid, high-throughput analysis of samples, making it ideal for genomic mutation or polymorphism screening studies in both clinical and research settings. PMID- 15884683 TI - Etiologic agents of dermatophyte infection in Lebanon. AB - BACKGROUND: Dermatophytes are common and cause important human fungal infections in many parts of the world, including Lebanon. The prevalence of these fungi, however, tends to vary with time and geographic location. In our region, studies on the prevalence of dermatophytoses and the distribution of the various dermatophyte species involved are rare. In Lebanon, only one study was published on this subject over the last forty years. OBJECTIVES: This study was undertaken to shed light on the types and prevalence of dermatophytes recovered at the American University of Beirut Medical Center (AUBMC), and to compare the findings with those of the only study from the same hospital published in the early 1960s. METHODS: Records from the clinical microbiology section were reviewed for the patients on whom dermatophyte cultures were requested between 1996 and 2002. All positive cultures were identified and analyzed. RESULTS: Among 1631 submitted specimens for culture (one per patient), 208 (12.7%) yielded 11 species of dermatophytes, dominated by Trichophyton spp. (89.9%), followed by Microsporum spp. (9.1%), Epidermophyton floccosum (0.4%) and Trichosporum beigelii (0.4%). The male to female ratio was almost 1:1, and the age range was 1 to 77 yrs for both sexes; 14% were children (< or = 15 yrs). The most commonly recovered species were: T. tonsurans (54.8%), T. mentagrophytes (24.5%), M. canis (7.7%), T. rubrum (5.3%) and T. verrucosum (4%). The distribution of infection according to body sites was : tinea unguium (44.2%), tinea corporis (43.2%), tinea capitis (7.7%) and tinea pedis (4.8%). Compared to the previous study from AUBMC in 1962, the current study showed an overall lower prevalence of dermatophytoses (12.7 vs 18.5%, p = 0.001) and variations in the prevalence rates of the dermatophyte species involved (dominated by T. tonsurans and T. mentagrophytes in the current study compared to E. floccosum and T. rubrum recovered in 1962). Based on the source of infection the current study showed higher transmission from human to human (anthropophylic) (86.5 vs 66.3%) and animal to human (zoophylic) (12.5 vs 1.1 %), but lower transmission from soil to human (geophylic) (1% vs 32.6%). CONCLUSIONS: The prevalence of dermatophytoses remains high in this country. The recognition of the changing prevalence in the causative dermatophyte species should help with the treatment approach and for potential implementation of control measures. PMID- 15884684 TI - Symptoms and acute pesticide intoxication among agricultural workers in Lebanon. AB - Pesticides are known for their toxicity and are still widely used, especially in agriculture. Increasing attention is being focused on the health effects affecting humans occupationally exposed to pesticides, particularly agricultural workers. The objective of this study was to assess the association between occupational exposure to pesticides with recurrent symptoms and acute intoxications, in comparison with other workers in Lebanon. A cross-sectional study was undertaken, where a standardized questionnaire was administered to 89 exposed and 88 non exposed workers. Subjects exposed to pesticides reported more upper respiratory tract infections (PR = 9.39 ; p < 10(-4)), anorexia (PR = 3.63 ; p = 0.03), epistaxis (PR = 2.97; p = 0.02), immunoallergic (PR = 1.58 ; p = 0.01) and gastrointestinal (PR = 2.13 ; p < 10(-4)) symptoms. Acute pesticide intoxications requiring hospitalization were also more common (PR = 4.26; p = 0.01). Adjustment over potential confounding did not change the results, except for the appearance of an association of central (PR = 1.50 ; p = 0.04) and autonomic nervous systems symptoms (PR = 3.92 ; p = 0.01) with agricultural work in Mount Lebanon. Our results suggest that agricultural workers have a higher prevalence of multiple symptoms, which may be due to subacute intoxications by pesticides that did not need hospitalization. In addition, these workers have a higher risk of having an acute intoxication due to pesticides, exposing them to life-threatening situations. PMID- 15884685 TI - [Typhoid fever. Clinical and therapeutic study in 70 patients]. AB - OBJECTIVE: Present clinical features of typhoid fever in Lebanon. METHODS: Retrospective study of 70 patients admitted to a university hospital center between 1995 and 2002. The criteria for inclusion were a positive Salmonella typhi or paratyphi hemoculture and/or a Widal serodiagnosis > or = 1/160 for O agglutinin, in the presence of evocative symptoms. RESULTS: Among the 70 patients, 25 pediatric cases were noted. The patients were aged a mean of 28+/-22 years. Average duration of symptoms before the diagnosis was 10+/-7 days. Fever were observed in 97% of cases and the other predominant symptoms were abdominal pain (41%), diarrhoea (36%), chills (31%) and headache (29%). Febrile gastroenteritis was a frequent manifestation in children (52%). Complications were noted in 31% of cases and were predominantly digestive. Leucopenia was not a helpful diagnostic marker. S. typhi was the most frequent (87%) serotype identified. Resistance to ampicilline was 10%, to cotrimoxazole and chloramphenicol 7% for each and to ofloxacine 2%. One death was reported (1%) of an immunosuppressed patient. CONCLUSION: Typhoid fever is still an endemic disease in Lebanon and should be systematically evoked in the case of prolonged fever, febrile gastroenteritis and/or headache. The appearance of bacteria resistant to antibiotics makes ceftriaxone or ciprofloxacine the empirical treatment of choice. PMID- 15884686 TI - THE LESSA (Lebanese Short-term Study with Amisulpride) STUDY a Phase IV clinical trial in acutely ill schizophrenic patients. AB - BACKGROUND: Conventional antipsychotics have relatively moderate efficacy against schizophrenic negative symptoms and are associated with significant adverse events. OBJECTIVES: To assess the clinical safety and efficacy of Solian (amisulpride), an atypical antipsychotic agent of the benzamide family. PATIENTS AND METHODS: This is an 8-week cohort study including adults with schizophrenia. Daily dose was between 400 and 800 mg. Safety was measured as the cumulative risk of adverse events and efficacy as the physician's global assessment of global improvement on an ordinal scale from 0 to 6. An efficacy index, a composite measure of both safety and efficacy, was also measured on an ordinal scale from 16 the worst to 1 the best. RESULTS: Fifty-seven patients were included. Eight patients (14%) experienced at least one adverse event during the trial, mainly extra-pyramidal symptoms (EPS). None required treatment discontinuation and none was qualified as serious. Four required corrective treatment and six resolved during the 8-week follow-up. On the Global Improvement scale 31.6% of patients were "very much" or "much improved" on D14 and 81.8% on D56. 25.5% of subjects achieved complete remission over the study period. Improvement reflected also on the efficacy index, decreasing from 8.2 (SD : 3.1) on D14 to 3.4 (SD: 2.9) on D56. CONCLUSION: Amisulpride, in addition to its well-established efficacy in schizophrenic patients, offers a particularly good safety profile. PMID- 15884687 TI - Fibromyalgia in Iraqi patients with Behcet's disease. AB - OBJECTIVE: To determine the prevalence of fibromyalgia syndrome (FMS) in Iraqi patients with Behcet's disease. METHODS: The two-stage classification process proposed by the 1990 American College of Rheumatology (ACR) multicenter criteria committee on fibromyalgia was applied after completion of a questionnaire about Behcet's disease clinical features. Stage I: A plain questionnaire to a sample of 90 (41 female and 49 male) patients with Behcet's disease, mean age 37.5 years. Stage II: All patients with wide spread pain were examined by a physician blinded to the diagnosis of Behcet's disease for 18 tender points and 4 control non tender points. A sample of 40 (16 female and 24 male) healthy individuals were examined as controls. RESULTS: There were 53 (58.9%) patients with BD with widespread pain compared to 6 (15%) individuals in the control group, whereas only 8 (8.9%) (7 female and 1 male) patients with BD fulfilled the 1990 ACR criteria for classification of FMS, compared to one individual in the controls (2.5%). There were significantly more females with BD having FMS compared to male patients. Also the FMS associated features were more prevalent in BD group compared to control group. CONCLUSIONS: FMS and widespread pain are more prevalent in patients with BD than in the general population while it is less prevalent when compared with other connective tissue diseases. There is a trend for an increased frequency of FMS in females with BD. BD patients with FMS may benefit from psychological evaluation as a part of their treatment. PMID- 15884688 TI - [Pulmonary Langerhans-cell histiocytosis in adults]. AB - Pulmonary Langerhans'cell histiocytosis belongs to the spectrum of the Langerhans'cell histiocytosis. The common point is tissue infiltration by Langerhans' cell granuloma. But the pulmonary type occurs predominantly in young adults and is associated with tobacco. The most frequently encountered symptoms are cough and dyspnea. Twenty-five to 35% of the cases are incidental findings. 20% of cases present with extra-thoracic lesions including skin, bone and hypothalamo-pituitary axis. On scanning images the lung parenchyma is infiltrated at the beginning by stellar nodules of less than 10 mm, which evolve either gradually to form cysts or towards remission, spontaneous or induced by the treatment. With pulmonary function testing, the two types of anomalies (obstruction and restriction) are observed. The reduction in diffusing capacity observed in 60 to 90% of the cases reflects the pulmonary vascular involvement. Detection of more than 5% of CD-1a cells in bronchioloalveolar lavage fluid and transbronchial biopsies are not sensitive. Treatment is controversial and includes primarily smoking cessation and corticosteroid therapy at the early nodular stage. PMID- 15884689 TI - The role of inflammation in acute coronary syndromes: review of the literature. AB - Extensive evidence supports a pathogenic role for both local and systemic inflammation in acute coronary syndrome (ACS). The determination of inflammatory markers levels in combination with the new imaging modalities can be used to identify patients at high risk of future cardiovascular events. Interestingly, this inflammatory component is a novel therapeutic target in ACS. PMID- 15884690 TI - [Adenocarcinoma of the gastro-esophagus junction associated to endobrachyesophagus. Two case reports. Review of the literature]. AB - Barrett esophagus (BE) is intestinal metaplasia (MI) within the distal tubular esophagus. The BE results in replacement of the normal squamous-lined epithelium with a columnar type epithelium. This metaplastic lesion is a clearly defined risk factor for the development of esophageal adenocarcinoma (ADC). In the western countries the incidence of adenocarcinoma of the lower esophagus and the gastro-esophagus junction have rapidly increased during the past twenty years. The 5-year survival is very poor. Although the relative risk of individuals in the United States with BE developing esophageal adenocarcinoma is very high, the absolute risk is extremely low due to the small number of cases. This lesion is caused by a persistent gastro-esophageal reflux. The nature of the reflux liquid is mixed acid and alkaline in the big majority of cases. A familial aggregation of BE and esophageal adenocarcinoma are present in 14% of patients with BE and esophageal adenocarcinoma. The diagnosis, the surveillance, the new tools of characterization of BE and the therapy remain an actual problem. We present 2 cases of endobrachyesophagus associated to an adenocarcinoma of the lower esophagus and a review of the main actual problem. PMID- 15884691 TI - Cutaneous aspergillosis in a renal transplant recipient. AB - Aspergillosis is a serious infection in renal transplant patients especially if dissemination occurs. We report a case of primary cutaneous aspergillosis, an extremely rare entity described in only four cases in renal transplant. It should be sought when surgical wound shows a rapidly growing necrotizing features early post transplant and without evidence of hematogeneous or contiguous tissue or organ involvement. Early suspicion, diagnosis, extensive surgical debridement as well as rapid institution of Amphotericin B can minimize the risk of dissemination. PMID- 15884692 TI - Spontaneous pneumothorax following chemotherapy for metastatic germ cell tumor: a case report. AB - We report on a 30-year-old man with metastatic non seminomatous germ cell tumor of the left testicle to the abdomen and the lungs, who suddenly developed a bilateral spontaneous pneumothorax after the first course of salvage chemotherapy. Rapid destruction and lysis of lung nodules by chemotherapy seem to be the main mechanism of pneumothorax development. According to our case report and to the literature, the onset of acute dyspnea after chemotherapy for lung metastatic germ cell tumor should alert to the possibility of spontaneous pneumothorax. PMID- 15884693 TI - [Image of the month. Diagnosis of micropolycystic ovary by transvaginal ultrasonography]. PMID- 15884694 TI - [Image of the month. Massive intramediastinal aortic rupture]. PMID- 15884695 TI - [How I treat ... atopic dermatitis by topical pimecrolimus (Elidel). The emerging paradigm of calcineurin inhibitors]. AB - Topical calcineurin inhibitors, also called topical immunomodulators or downregulators, represent an innovative class of non-steroidal anti-inflammatory agents. Pimecrolimus 1% cream (Elidel) is one representative drug available for the treatment of atopic dermatitis. Unlike topical steroids, this drug does not affect collagen synthesis and does not alter the dendritic cell functions and the barrier function of the skin. PMID- 15884696 TI - [Replacement of the descending aorta sparing the aortic valve: the David's operation]. AB - We report the case of an ascending aortic aneurysm involving the aortic root, with a functionally competent bicuspid aortic valve, treated by the David's operation. This operation allows to fix the aortic root pathology, whilst preserving the native aortic valve. The technical aspects, surgical indications, and potential benefits of this operation are reviewed. PMID- 15884697 TI - [Penile verrucous carcinoma]. AB - Penile verrucous carcinoma is a rare tumor. Verrucous carcinoma is thought by most to represent a well-differentiated or low-grade squamous-cell carcinoma. The term Buschke-Lowenstein tumor and giant condyloma have also been used to identify this histological lesion. A review of literature points to the role of human papillomavirus infection. The authors report a case of a penile verrucous carcinoma in a circumcised man. PMID- 15884698 TI - [Endoscopic mucosa resection for superficial esophageal carcinoma: an alternative to radical surgery?]. AB - Radical surgery is the standard of care for resectable esophagus cancers, with mortality less than 5% in heavily trained surgical teams. Nevertheless, the morbidity of surgery is quite high due to the procedure and due to the poor status of these patients. Endoscopic mucosal resection seems effective for the resection of superficial malignant lesions of esophagus with an acceptable morbidity profile. A correct staging has to confirm the superficial character of the lesion and exclude deeper infiltration into the digestive wall or nodal involvement. This technique might be an acceptable alternative to surgery or superficial lesions, particularly for patients at high risk for aggressive surgical procedure. PMID- 15884699 TI - [Guidelines on cardiovascular disease prevention in clinical practice]. AB - These recommandations are largely based on the "European Guidelines on Cardiovascular Disease Prevention in Clinical Practice" proposed by the "Third Joint Task Force of European and other Societies on Cardiovascular Disease Prevention in Clinical Practice". The model used to assess the overall risk was adapted for Belgium. Otherwise, very few things were changed from the Exectutive Summary of the European Guidelines. PMID- 15884700 TI - [Clinical evaluation of 23 children with neuroblastoma. The experience of a single institution]. AB - In this retrospective study, we analyse epidemiology, clinical symptoms and therapeutic results in a group of 23 children with neuroblastoma. Half of them were less than 2 years of age; in 19 of 23, the primitive tumour was abdominal; 35% of them were initially metastatic. The overall survival was 83% at 5 years and the event free survival, 75% at 5 years. Pronostic factors are age, extension of the disease at diagnosis, biologic parameters and genetic study of the neuroblast cells (amplification of N-myc oncogen). Our study shows the deleterious effect of bone lesions. PMID- 15884701 TI - [Psychotropic drug use in the Belgian non-institutionalized population]. AB - OBJECTIVE: To assess psychotropic drug utilization in the general population of Belgium, and the pattern of use in individuals with mental disorders. METHOD: A representative random sample of non-institutionalized inhabitants from Belgium aged 18 or older (n = 2419) were interviewed between April 2001 and June 2002. Individuals were asked about any psychotropic drug use in the past 12 months, even if they used the drug(s) just once. The interview took place by means of the World Mental Health Composite International Diagnostic Interview (WMH-CIDI). RESULTS: Slightly more than 19% reported that they had used any psychotropic drug in the past 12 months; 11% had a 12-month mental disorder. The extent of psychotropic drug utilization varied according to the nature of the disorders: 60% in mood disorders, 53% in alcohol disorders, and 39% in anxiety disorders. Thirty nine percent of the respondents with a mood disorder consumed an antidepressant drug in the past year; 23% of the respondents with an anxiety disorder consumed an anxiolytic drug in the past 12 months. Among those who reported that they used antidepressants, selective serotonin reuptake inhibitors were most commonly consumed (SSRI). CONCLUSION: These data do not provide evidence for any over-consumption of psychopharmacological agents in Belgium. By contrast, they question the appropriateness of pharmacological treatments, in which under-treatment seems to be coupled with a high use of non-specific medications. PMID- 15884702 TI - [News about phosphorus metabolism]. AB - Phosphorus is mainly present in the serum as phosphate which is the principal intracellular anion, essential for the organism, in particular through its role in the production of ATP. In plasma, only inorganic phosphorus (Pi) is measured. In this article, we will review new information about the regulation of Pi at the intestinal, osseous and mainly renal levels. The various cotransporters sodium phosphorus (Na-Pi) will also be examined and the molecular regulation mechanisms approached. Finally, illustration of Pi physiology will be given through various clinical examples. PMID- 15884703 TI - [Aspirin and cardiovascular prevention: last minute information]. AB - Just a few days after the publication in this journal of a review on aspirin and cardiovascular prevention, a significant article appeared in the international literature; it provides new information and deserves a brief presentation. This paper is concerned with patients who took aspirin to prevent vascular diseases and who presented with ulcer bleeding. After the ulcers had healed and after eradication of Helicobacter pylori had, if necessary, been achieved, 320 patients were randomly assigned to receive either 75 mg of clopidogrel daily or 80 mg of aspirin daily + 20 mg of esomeprazole twice daily. Recurrent ulcer bleeding occurred in 13 of the 161 patients assigned to receive clopidogrel and in 1 of the 159 who received aspirin plus esomeprazole. The cumulative incidence of recurrent bleeding during the 12 months of follow up was 8.6% in the clopidogrel group and 0.7% in the aspirin-esomeprazole group (p = 0.001). These findings do not support a current American recommendation that patients with major gastrointestinal intolerance of aspirin should be given clopidogrel instead. PMID- 15884704 TI - Exploring virtual environments with an EEG-based BCI through motor imagery. AB - In this paper, we describe the possibility of navigating in a virtual environment using the output signal of an EEG-based Brain-Computer Interface (BCI). The graphical capabilities of virtual reality (VR) should help to create new BCI paradigms and improve feedback presentation. The objective of this combination is to enhance the subject's learning process of gaining control of the BCI. In this study, the participant had to imagine left or right hand movements while exploring a virtual conference room. By imaging a left hand movement the subject turned virtually to the left inside the room and with right hand imagery to the right. In fact, three trained subjects reached 80% to 100% BCI classification accuracy in the course of the experimental sessions. All subjects were able to achieve a rotation in the VR to the left or right by approximately 45 degrees during one trial. PMID- 15884705 TI - [Cell proliferation and cellular activity of primary cell cultures of the oral cavity after cell seeding on the surface of a degradable, thermoplastic block copolymer]. AB - Using standard cell biological and biochemical methods we were able to test the ability of a degradable, thermoplastic block copolymer to support the adhesion, proliferation, and the cellular activity of primary cell cultures of the oral cavity in vitro. The delicate balance between a group of endogenous enzymes, Matrix Metalloproteinases (MMPs), and their inhibitors (Tissue Inhibitor of MMPs, TIMPs) have a decisive function in the remodeling of the extracellular matrix during processes like wound healing or the integration of biomaterials in surrounding tissues after implantation. Recently developed, biodegradable thermoplastic elastomers with shape-memory properties may be the key to develop new therapeutical options in head and neck surgery. Primary cell cultures of the oral cavity of Sprague-Dawley rats were seeded on the surface of a thermoplastic block copolymer and on a polystyrene surface as control. Conditioned media of the primary cells were analyzed for MMPs and TIMPs after different periods of cell growth. The MMP and TIMP expression was analysed by zymography and a radiometric enzyme assay. No statistically significant differences in the appearance and the kinetic of MMP-1, MMP-2, MMP-9 and TIMPs were detected between cells grown on the polymer surface compared to the control. An appropriate understanding of the molecular processes that regulate cellular growth and integration of a biomaterial in surrounding tissue is the requirement for an optimal adaptation of biodegradable, polymeric biomaterials to the physiological, anatomical, and surgical conditions in vivo to develop new therapeutic options in otolaryngology and head and neck surgery. PMID- 15884706 TI - [Electrochemical properties of biocompatible material hardness modifications on titanium and steel under mechanical loads]. AB - Friction corrosion may appear between different implant components or between implant and hard tissue. The sliding micro movements induce fretting wear corrosion and have been recently reported as a cause of joint prostheses failure. A surface coating is desirable, that retains the mechanical properties of the substrate, offers good biocompatibility and improves the fretting corrosion resistance. In this study it could be demonstrated that tantalum and niobium coatings fulfill the requirements. On titanium substrates the coating decreases the abrasion against PMMA, an orthopedic relevant material. Furthermore, in the case of medical steel substrates the biocompatibility and the corrosion properties are improved. The better abrasion-resistance is minimizing the release of allergological critical particles like nickel and chromium. PMID- 15884707 TI - The critical size bony defect in a small animal for bone healing studies (I): Comparative anatomical study on rats' femur. AB - Laboratory rats are small animal models which are often used for scientific investigations in medicine. So far there are only few scientific data about the meaning of these small animal models for in vivo bone healing studies available in literature. Although the rat's femur with its cyclic loadings during gait is an appropriate model for investigations in the field of experimental orthopaedics and traumatology there is a lack of morphometric information with respect to its anatomy. The aim of this study is to evaluate the anatomy of rat femurs in two species, which are often performed in animal experimental medicine. These morphometric data should contribute to develope an appropriated osseous fragment fixation system in the rat's femur. The femurs of Wistar (WR) and Sprague Dawley (SDR) cadavers were prepared and analysed by x-rays in two standard planes. The results were compared with the corresponding data for humans by literature. It could be demonstrated that SDR showed a higher caput-collum-diaphyseal and antetorsion angle, but a lower transcondylar femur valgus angle compared to WR. Cortical thickness, bone marrow cavity diameter and femur length were higher in WR. Wistar rat's femur anatomy shows more similarities to human anatomy than Sprague Dawley rats. PMID- 15884708 TI - [Application of a stand-alone interbody fusion cage based on a novel porous TiO2/glass ceramic--2: Biomechanical evaluation after implantation in the sheep cervical spine]. AB - Animals are becoming more and more common as in vivo models for the human spine. Especially the sheep cervical spine is stated to be of good comparability and usefulness in the evaluation of in vivo radiological, biomechanical and histological behaviour of new bone replacement materials, implants and cages for cervical spine interbody fusion. In preceding biomechanical in vitro examinations human cervical spine specimens were tested after fusion with either a cubical stand-alone interbody fusion cage manufactured from a new porous TiO2/glass composite (Ecopore) or polymethylmethacrylate (PMMA) after discectomy. Following our first experience with the use of the new material and its influence on the primary stability after in vitro application we carried out fusions of 20 sheep cervical spines levels with either PMMA or an Ecopore-cage, and performed radiological examinations during the following 2-4 months. In this second part of the study we intended the biomechanical evaluation of the spine segments with reference to the previously determined morphological findings, like subsidence of the implants, significant increase of the kyphosis angle and degree of the bony fusion along with the interpretation of the results. 20 sheep cervical spines segments with either PMMA- or Ecopore-fusion in the levels C2/3 and C4/5 were tested, in comparison to 10 native corresponding sheep cervical spine segments. Non-destructive biomechanical testing was performed, including flexion/extension, lateral bending and axial rotation using a spine testing apparatus. Three dimensional range of motion (ROM) was evaluated using an ultrasound measurement system. In the native spine segments C2/3 and C4/5 the ROM increased in cranio caudal direction particulary in flexion/extension, less pronounced in lateral flexion and axial rotation (p < 0.05). The overall ROM of both tested segments was greatest in lateral flexion, reduced to 52% in flexion/extension and to 16% in axial rotation. After 2 months C2/3- and C4/5-segments with PMMA-fusion and C2/3-segments with Ecopore-interposition showed decrease of ROM in lateral flexion in comparison to the native segments, indicating increasing stiffening. However, after 4 months all operated segments, independent from level or implanted material, were stiffer than the comparable native segments. The decrease of the ROM correlated with the radiological-morphological degree of fusion. Our evaluation of the new porous TiO2/glass composite as interbody fusion cage has shown satisfactory radiological results as well as distinct biomechanical stability and fusion of the segments after 4 months in comparison to PMMA. After histological analysis of the bone-biomaterial-interface, further examinations of this biomaterial previous to an application as alternative to other customary cages in humans are necessary. PMID- 15884709 TI - [Bionic surface design in metal on metal bearings for total hip arthroplasty- optimization of tribological characteristics]. AB - Bionic systems are aiming to integrate natural observing into mechanical solutions. This has been realized in the design of metal on metal bearing in total hip resurface arthroplasty. The articular side of the femoral cup is covered with a dimple like surface. Under laboratory condition this so called "surf-metal-cup" achieved a reduction of the mechanical wear to almost a third part in comparison to a metal-cup with plane surface. This advantage, caused by the reduced friction-coefficient due to improved hydrodynamic lubrication could also be proved under laboratory conditions. The clinical introduction is expected to offer a significant extension of durability in this prosthetic system and needs to be proved in a long-term study. PMID- 15884710 TI - [Esophageal pH monitoring study of eight hours in pediatric patients with suspected gastroesophageal reflux]. AB - PURPOSE: To study the pH monitoring over eight hours, analyzing variables such symptomatology and esophagogastroduodenoscopy, as well as the following pH monitoring parameters: number of reflux episodes, number of episodes longer than five minutes, length of reflux time during the eight hours, and the episode of longest duration, for the diagnosis of gastroesophageal reflux. METHODS: A prospective study. Continuous measurement of esophageal pH was performed over eight hours on 35 patients whose ages ranged from four months to 11 years. The DPS-100 Diagnosis pH meter System was utilized, programmed in accordance with the methodology suggested by Vandenplas. The pH monitoring results were divided into positive and negative and compared with all the variables and parameters of the examination. RESULTS: We obtained statistically significant differences in all the pH monitoring parameters analyzed, with the reflux index being the most effective for identifying the pathological reflux, independent of age. None of the qualitative variables of age, symptomatology and esophagogastroduodenoscopy showed statistically significant differences when compared to the pH monitoring results. CONCLUSION: The esophageal pH monitoring of eight hours has its pH monitoring parameters validated for the diagnosis of gastroesophageal reflux. PMID- 15884711 TI - Acetylcysteine in random skin flap in rats. AB - PURPOSE: Analyze the ability of Acetylcysteine to reduce distal necrosis in a random skin flap, in the rat. METHODS: The present study utilized 28 adult male Wistar-EPM rats distributed, at random, in two groups of 14 animals. Control group rats (CG) received distilled water and Acetylcysteine group animals (NACG) received NAC (300 mg/kg) by oral infusion, 15 minutes before flap elevation. On the seventh postoperative day, percentage of distal necrosis was determined and skin samples collected in order to allow determination of MDA levels. RESULTS: The mean necrotic area in CG group (control) was 66% and in NACG group (Acetylcysteine) 52%, a statistically significant difference according to the Mann-Whitney test (U calc = 25; U crit = 45). MDA levels were lower in the CG flap skin samples than in the NACG samples (U calc = 24; U crit = 45), the oposite being true in the normal skin samples (U calc = 10; U crit = 45). CONCLUSION: Acetylcysteine was effective, according to the model used, reducing the percentage of distal necrosis in NACG rats. PMID- 15884712 TI - Effects of aging on abdominal wall healing in rats. AB - PURPOSE: The aim of this study was to assess abdominal wall healing in old and young adult rats. METHODS: On average, young animals were 110 days old and old animals were 762 days old. A 4.0 cm median laparotomy was performed under anesthesia, followed by laparorrhaphy on two synthesis planes, i.e. peritoneum muscle-aponeurosis and skin, using continuous 5.0 nylon sutures. The animals were evaluated on the 3rd, 7th, 14th and 21st postoperative days. The resistance of the two planes was studied separately and a histopathologic analysis was performed on sections stained with hematoxylineosin and Sirius Red. Immunohistochemical analysis was also carried out using PCNA, LCA and CD34. RESULTS: The skin scars gained resistance in a similar manner at the initial time points, but those of young rats were more resistant on the 21st day (p=0.0029). Total and type III collagen content was similar in the two groups and type I collagen content was higher in young animals on the 14th day. Inflammatory cell infiltration was more marked in the skin wounds of young animals on the 3rd day (p=0.0190). Reepithelialization was similar and angiogenesis was more intense in the skin wounds of young animals on the 14th day (p=0.0062). The peritoneum muscle-aponeurosis wounds gained similar resistance during the early phases, but were more resistant on the 14th day (p=0.0005) and on the 21st day (p=0.0023) in old rats Collagen concentration was higher in the wounds of old animals on the 3rd day (p=0.0112) and in the wounds of young animals on the 21st day (p=0.0348). The inflammatory reaction was more intense in the wounds of old animals on the 3rd day (p=0.0060) and angiogenesis was more intense on the 14th day (0.0432). CONCLUSION: Although there are some differences in the healing course between young and old animals, age, of itself, does not impair the healing of abdominal wall wounds in rats. PMID- 15884713 TI - [Oxidative alterations, total antioxidant status and nitric oxide study in rats submitted to ischemia and reperfusion of hind limbs]. AB - PURPOSE: To verify the effect of the ischemia time on the oxidative alterations, total antioxidant status and the nitric oxide, in the muscle, kidney and plasma, in rats submitted to ischemia and reperfusion of hind limbs. METHODS: Forty male rats were randomized in four groups with ten animals each. Laparotomy, abdominal aorta infra ejuxta renal isolation. Groups 1 and 3 (sham), passage of the polipropilen 7.0 monofilamentary thread aorta around without bending it and waiting time of 1 hour to group 1 and 6 hours to group 3. Withdrawal of the thread, waiting time of 15 minutes, euthanasia and material collection. Groups 2 and 4 (experiments), ligation of abdominal aorta with the same thread and ischemia of 1 hour in the group 2 and 6 hours to the group 4. Withdrawal of the thread and reperfusion for 15 minutes. Euthanasia and blood collection by flowing; withdrawal of the left hind limb muscle and of the left kidney. The parameters evaluated were: malondialdehyde, total antioxidant status and the nitric oxide. RESULTS: The "T" Student test and the MANN-WHITNEY test have been used as nonparametrics tests and the rejection level have been fixed in 0.05 or 5%. There were significative alterations of the malondialdehyde in the kidney. The total antioxidant status has been shoun significative alterations in the kidney, muscle and plasma. Although the nitric oxide has been altered in the plasma only. CONCLUSION: The ischemia time changing has provoked significative alterations of malondialdehyde, total antioxidant status and nitric oxide. PMID- 15884714 TI - [Effects of tenoxicam on abdominal wall healing: experimental study in rats]. AB - PURPOSE: To analyse the effect of tenoxicam, a nonsteroidal anti-inflammatory, on the abdominal wall healing in rats. METHODS: It was used 40 rats, submitted to longitudinal laparotomies, and allocated, randomly, in one control group (C), consisted of 20 rats treated with saline solution; and one test group (T), consisted of 20 rats treated with tenoxicam. The animals of each group were divided, according to their sacrifice day, into subgroups of 10 animals, named as C7, C14, T7 and T14. The numbers 7 and 14 indicated that the animal would be sacrificed on the 7th and 14th postoperative day, respectively. The tenoxicam (1 mg/ml) and saline solution (NaCl 0.9%) were administrated by intramuscular injections, at the dose of 0.6 ml/kg/day, immediately after surgery and continued for 4 days. In the sacrifice day, two segments of the abdominal wall (1 cm x 3 cm) were prepared and submitted to breaking strength measurement and hydroxyproline determination. RESULTS: No complications were observed in the four subgroups, including infection or dehiscence. There were no significant differences in the breaking strength measurement (p=0.262) and the hydroxyproline levels (p=0.392) among the four subgroups. CONCLUSION: The tenoxicam, administered intramuscularly, does not interfere on the abdominal wall healing of rats. PMID- 15884715 TI - [Histologic analysis of colonic anastomotic healing, in rats, under the action of 10% Aroeira-do-sertao (Myracrodruon urundeuva fr. all.) enema]. AB - PURPOSE: Evaluate, from a histologic view point, the action of the aqueous extract of aroeira-do-sertao on the healing of colonic anastomosis, in Wistar rats. METHODS: There were used 48 Wistar rats, males, with average weight of 320g, distributed in two groups, with 24 animals, each. All animals were subjected to a complete transverse section of the descending colon, followed by colonic anastomosis. The rats on group A received daily post-operative enemas of carboxymethylcellulose (CMC) based vehicle. The animals on group B received post operative enemas of 10% aqueous extract of aroeira-do-sertao in CMC based vehicle, until the date of euthanasia. On days 3, 7, 14 and 21 of the experiment, six rats in each group were subjected to removal of a colonic segment, including the anastomosis, destined to histological evaluation and qualitative analysis of inflammatory and healing cell response. RESULTS: The morphologic analysis reveals a significant difference between groups, on day 7 (n < 0.05), when the group studied showed lower score than the control group. The collagen deposition on the aroeira group was lower, on days 3, 7 and 14 (n < 0.05), compared to the control group. CONCLUSION: The aqueous extract of aroeira-do-sertao has anti-inflammatory action. It also has inhibitory effect on collagen deposition during the phases of inflammation and fibroplasia of the healing process of colonic anastomoses, in Wistar rats, but in advanced phases (day 21) the healing process is similar to that in the animals of the control group. PMID- 15884716 TI - [Comparative study of the hyperbaric hyperoxygenation in ischemic colonic loops in rats]. AB - PURPOSE: To analyze and to evaluate the effect of the hyperbaric hyperoxygenation in the mechanical resistance of ischemic colon of rats. METHODS: Eighty rats, distributed in four groups of 20 animals in each one, were used. In group 1 (G1), the control group, ischemia was not caused. Group 2 was submitted to the lesser degree of ischemia. Group 3 was submitted to the intermediate degree of ischemia. In group 4, a bigger degree of intestinal ischemia was provoked. Each group was divided in two sub-groups often animals each: with hyperbaric chamber (CC) and without hyperbaric chamber (SC). The animals of the four CC subgroups were placed in an experimental hyperbaric chamber in order to inhale oxygen at 100%, at two Absolute Atmospheres, for 120 minutes, for a four-day consecutive period. The animals of the four SC subgroups were kept in environment air during the five days of the experiment. All animals have been submitted to the mechanical study of the intestinal loop by the pressure test of the rupture by liquid distension. The euthanasia occurred in the fifth post-operative day. RESULTS: Considering the ischemia factor, the four groups were different among them (p=0.0001). There was no statistical difference between subgroups CC and SC (p=0.3461). CONCLUSION: The hyperbaric oxygen-therapy did not present improvement on the induced ischemia in rats upright colic loop. PMID- 15884717 TI - [Internal cardiac massage in dogs: a new technique proposition for emergency pericardiotomy--ligament traction]. AB - PURPOSE: Describe a technique of emergency pericardiotomy, named as Ligament Traction (LT), to reduce the necessary time to begin the Internal Cardiac Massage. To perform the ICM an emergency toracotomy and pericardiotomy are necessary, both in remote time. The technique usually employed is the "T" pericardiotomy, whose execution depends on the apprehension of the pericardium with an Allis forceps. This apprehension is difficult and complicates the reanimation of the patient. METHODS: Twenty canine corpses were divided into two groups: Group I--"T" pericardiotomy (n=10), and Group II--the LT technique (n=10). The LT consisted on the traction of the pericardiumphrenic ligament and the section of the pericardium next to its apex. The incision was elongated with the introduction of the fingers, also allowing the positioning of the heart in the hand of the operator and the immediate beginning of the ICM. RESULTS: Group I presented an execution period of 21.79 +/- 0.88 second, and Group II of 8.58 +/- 1.38, with p<0.0001 (highly expressive). CONCLUSION: The technique of pericardiotomy by Ligament Traction concur to outliving, because it avoids a larger time of cerebral ischemia, due to the early beginning of the circulation. PMID- 15884718 TI - [Topical administration of hydralazine hydrochloride on the viability of randon skin flaps in rats]. AB - PURPOSE: Assess the effect of hydralazine hydrochloride, for iontophoresis, on the viability of random skin flaps in rats. METHODS: Sixty Wistar rats was randonly destributed in 4 groups (n=15), these animals was submited as randon dorsal skin flaps as cranial base with measure 10 x 4 cm. The animals from group 1 was utilized as control, in group 2 was submitted to direct current o 4mA-20' immediately after the surgery and on the two subsequent days. In group 3 the stimulation eletric simulation with hydralazine hydrochloride. In group 4 iontophorese with hydralazine hydrochloride 4mA-20'. The analysis of the results was made on the seventh day post operative and interpreted with test non parametric of Kruskal-Wallis. RESULTS: and the necrotic area stayed fixed in: group 1=45%; group 2=39%; group 3=46% and group 4=41%, being the statistical analysis did not evidenced any significant. CONCLUSION: The hydralazine hydrochloride when taken for iontophorese was not efficacious in reduce the necrotic area. PMID- 15884719 TI - [One layer sutures of digestive tract knotted in the lumen, in dogs: perforating stitch versus serosubmucosal suture]. AB - OBJECTIVE: To compare hand sewn digestive tract single layer anastomosis with knots tied in the lumen: total stitches versus serosubmucosal. METHODS: Six mongrel dogs were submitted to laparotomy, each one with two transversal jejunum sections, 30 and 70 cm far from Treizt angle and suture, serosubmucosal and total stitches, both with knots tied in the lumen, over the mucosa, at the posterior wall. After slaughter (7th post-operative day) was evaluated the peritoneal adhesions at posterior wall. The macro and microscopic features was observed. Wilcox on rank sum test was applied for the histhometry. RESULTS: More profuse adhesions with the serosubmucosal stitches tied in the lumen with adherence tissue over the suture line, avoiding the serosa, within or without healing deformation of the suture lines, doing an anastomosis angle. There was good serosa reconstitution with total stitches. The epithelium was perfectly reconstituted at serosubmucosa, but not at total stitches, where was residual focus of acute inflammation. The reline and regeneration of wall components (except the serosa, whose regeneration was impaired by peritoneal adherences) were better with serosubmucosal then total stitches. The muscularis never regeneration in anyone suture. The polimorphonuclear cells, macrophages, fibroblasts, and collagen fibers was more numerous (statistical significance) at total stitches. CONCLUSION: Total stitches with knots tied in the lumen, at posterior wall, over the mucosa are safe full, despite of major inflammation. Serosubmucosal with knots tied in the lumen, at posterior wall, over the mucosa, allows peritoneal adherences formation, and should be avoided. PMID- 15884720 TI - [Study about the feasibility of ovarian graft in rats' omentum pouch, using Cyclosporine A]. AB - PURPOSE: To study comparatively ovarian autograft and heterograft in rats' omentum pouch, using Cyclosporine A. METHODS: In the present study 20 rats, distributed equally in four groups, had been used: Control Group (rats submitted to ooforectomy); Group A (ovarian autograft); group H (ovarian heterograft); e group C (ovarian heterograft and treatment with Cyclosporine A). All animals had been submitted to a bilateral ooforectomy. One of the removed ovaries had been individualized and grafted in Greater Omentum. The animals from group C received 10mg/kg/dia of cyclosporine subcutaneously during the period of the experiment. All animals had been submitted to a new surgical intervention after a period of 33 days for identification of the receiving bedstead and withdrawal of the grafted ovary, beyond the uterus and the vagina, followed of euthanasia. The parts had been submitted to the histological process of routine. RESULTS: All groups presented vaginal and uterine epithelium with one or two rows of simple cubical or cylindrical cells, spherical and central nucleus. It also had great inflammatory reaction in all grafts, with necrosis areas. At the 20th day of the experiment, one of the animals of group C died. CONCLUSION: It was concluded that the graft of the entire ovary in Greater Omentum of rats in this sample, independently of the use of Cyclosporine, did not revealed viable. PMID- 15884721 TI - [Tensional and morphologic analysis of the colonic anastomosis on 10% acetic acid induced colitis, in Wistar rats, treated with 10% aroeira-do-sertao (Myracrodruon urundeuva fr. all.) aqueous extract]. AB - PURPOSE: Verify, from a morphologic and tensional view point, the effect of the 10% aqueous extract of aroeira-do-sertao on the colonic anastomosis, during the occurrence of 10% acetic acid induced colitis, in Wistar rats. METHODS: There were used 48 Wistar rats, distributed in two groups. All animals were subjected to induction of colitis by aqueous solution of 10% acetic acid. Twenty-four hours later, the animals were subjected to laparotomy, transverse total colotomy and end-to-end anastomosis with 5-0 polypropylene. Group A (vehicle), animals treated with carboxymethylcellulose based vehicle. Group B (aroeira), animals treated with 10% aroeira aqueous extract. Both treatments were by means of enema. Groups A and B were distributed in subgroups A3, A7, A14 and A21; B3, B7, B 14 and B21, according to the foreseen euthanasia date (days 3, 7, 14 and 21). On these dates, the animals were relaparotomized, the colonic segment containing the anastomosis was ressected subjected to the pressure test and then to the, histological analysis. For the momphologic study, the slides were dyed with hematoxilin-eosine and the healing status was evaluated based on a score chart that ranked from 0 to 16. The higher the score, the better the healing. RESULTS: On the evaluation of the morphologic study, which quantifies the evolution and degree of healing, as a final result of the healing process, the group aroeira was superior to the group vehicle (p<0.05) on the twenty-first day, showing higher velocity on tissue repair that occurs by regeneration and not by fibrosis. With respect to the tension test, there was a statistically significant difference on day 3, with predominance of the group aroeira over the group vehicle (p<0.05). CONCLUSION: The 10% aroeira-do-sertao aqueous extract, has healing activity on the colonic anstomosis during the occurrence of 10% acetic acid induced colitis. It improves the resistance to tension on the colonic anastomosis zone on the 3rd day. PMID- 15884722 TI - [Functional and morphological study of the local and systemic hypothermia on dog's liver]. AB - PURPOSE: To compare hepatic lesions produced by two types of hypothermia; the systemic and the local or topic. METHODS: Twenty dogs distributed in two groups were studied: the first submitted to local hypothermia and the second to systemic hypothermia. In all groups, biochemical dosages for alanina allytransferase (A.L.T.), aspartate aminotrasnferase (A.S.T.) and direct bilirubin (T.D.), conventional optical microscopy and electronic transmission microscopy were performed in times T0, Test, and T60, that is, before the hypothermia (T0), after temperature stabilization at 10 degrees lower than initial temperature (Test), and after sixty minutes of hypothermia (T60). RESULTS: The data analysis, both of the biochemical profile and of the microscopy showed that in the group of animals with selective hypothermia, the hepatic lesions were more intense when compared to the systemic hypothermia group. CONCLUSION: The selective hypothermia causes more lesions to the liver than the systemic. PMID- 15884723 TI - [CME-ECG 5]. PMID- 15884724 TI - [Angiotensin II receptor blockers--evidence along the cardiovascular continuum]. AB - The introduction of Angiotensin II receptor blockers (ARB) in 1995 was another milestone in the pharmacological management of hypertension. Due to the manifold effects on several target organs Angiotensin II is one of the most important mediator in the pathogenesis of hypertension. The blockade of the Angiotensin II receptor type 1 is a crucial cornerstone in interrupting the pathophysiological pathways in hypertension. Furthermore ARB have an excellent tolerability comparable with placebo. In the last decade large placebo-controlled trials could prove the efficiency of ARB in terms of morbidity and mortality. Patients after acute myocardial infarction and patients with chronic heart failure benefit from treatment with ARB equally compared to treatment with ACE inhibitors. Combining ARB and ACE inhibitors in patient after myocardial infarction increases the rate of adverse events without improving survival. Increase of microalbuminuria and worsening of diabetic nephropathy is reduced by ARB in patients with diabetes type 2, but an advantage over ACE inhibitors could not be documented. Hypertensive patients with electrocardiographically left ventricular hypertrophy treated with ARB seem to have an additional benefit in terms of morbidity and mortality compared to treatment with beta-blockers. In the early treatment of stroke patients treated with ARB have a lower 12-mounth mortality than patients receiving placebo. In conclusion, Angiotensin II receptor blockers are due to their well proved efficiency, the cardio- and renoprotective qualities and the excellent tolerability profile a useful therapeutic option in the management of patients with hypertension. PMID- 15884725 TI - [Percutaneous vertebroplasty: a minimal-invasive procedure for pain treatment]. PMID- 15884726 TI - [Exercise-induced left arm pain and thrombocytosis]. AB - We describe the case of a 45-year-old male smoker who presented with an acute anterior wall myocardial infarction and a platelet count on admission of 1030000/mm3. Emergent coronary angiography revealead left anterior wall akinesia caused by a spontaneously resolved thrombosis of the left anterior descending artery with residual stenosis. Primary percutaneous coronary angioplasty and stenting were performed. Postangioplasty course was uneventful. He was diagnosed with essential thrombocythemia based on the findings of marked thrombocytosis of 1,030,000/mm3, splenomegaly and numerous clumping giant megakaryocytes on bone marrow biopsy. In addition to standard therapy with aspirin, heparin, betablocking agent, ACE-inhibitor and statine he received additional anti platelet therapy with Clopidogrel. Cytoreductive therapy was not necessary. PMID- 15884727 TI - [Right-sided headaches in a 50-year-old Turk]. PMID- 15884728 TI - [A neurological coincidence]. PMID- 15884729 TI - [CME-Ultrasonography 4/Solution]. PMID- 15884730 TI - George C. Williams: personal reminiscences. PMID- 15884731 TI - Red-letter days. PMID- 15884732 TI - Adaptation: "a critique of some current evolutionary thought". AB - In his classic Adaptation and Natural Selection: A Critique of Some Current Evolutionary Thought (1966), George Williams showed definitively that our understanding of adaptation, a central concept of evolutionary theory, must be gene-centered. The purpose of adaptations is to further the replication of genes. Genes are machines for turning out more genes; and adaptations are the means by which genes pluck resources from the world to promote this self-replication. Thus adaptations transform potential resources from part of the indifferent world-at large into tailor-made environments, environments brimming with resources for organisms' distinctive adaptive needs. Systematically dif ferent adaptive problems therefore give rise to different environments; and so different species, for example, have different environments. Thus a gene-centered analysis of adaptations implies a gene-centered theory of environments. Without genes to specify what constitutes an environment, environments would not exist. Rather than being separate from biology, an autonomous, independent force, environments are themselves the products of biology. So a gene-centered view, far from depreciating the environment, furnishes a rich and precise understanding of its importance. PMID- 15884733 TI - How do we decide that a species is sex-role reversed? AB - Defining sex roles has been driven by differences in mating systems at the extreme: polygyny and polyandry. Roles may reverse depending on which sex limits the reproductive rate of the other, and it is generally the female that limits the male. Males therefore compete for female mates. But in species in which the male limits the reproductive rate of the female, the female competes for male mates and assumes the masculine role. Complications arise, however, in species with typical roles when males are temporarily limiting, and females then briefly compete for and display to males. Problems also occur among tightly monogamous species with biparental care, where the mates have equal reproductive rates; both males and females compete intrasexually for mates. Despite this, monogamous species have masculine and feminine roles, typically manifested as the male dominating the female. Some monogamous species are nevertheless sex-role reversed. The pervasive behavioral mechanism characterizing the masculine role is dominance through aggression, size, or both. Attending more to behavioral mechanisms will enrich our understanding of sex-role reversal. PMID- 15884734 TI - The evolution of parental care. AB - Our understanding of parental care behavior can be significantly advanced through the application of Williams's Principle, which states that reproduction has not only a benefit but also a cost to lifetime fitness. My laboratory has formalized Williams's Principle into the relative value theorem and found that its application to fishes, the taxa with the most diverse patterns of parental care, can help to explain which sex provides care and how much. In fishes, it is often the male that provides parental care, not because the male obtains greater benefits from this care, but probably because he pays fewer costs. Fish dynamically adjust their investment into parental care according to the number of offspring in their brood, past investment, genetic relatedness, and alternative mating opportunities, all of which affect the value of current offspring relative to potential future offspring. These results may also help us understand the joy and the challenges of parental care in humans. PMID- 15884735 TI - The maintenance of sex as a developmental trap due to sexual selection. AB - The writings of George Williams challenged biologists to think critically about levels of selection, social behavior, and the paradox of sex, whose maintenance by recombination alone has not been convincingly demonstrated in theory or in fact. A solution is suggested by observations of the dependence of females on interaction with males, as a result of sexual selection. This, along with recombination in changing environments and DNA repair during meiosis, may contribute to a pluralistic explanation for the maintenance of sex. PMID- 15884736 TI - Carpe diem: adaptation and devaluing the future. AB - Organisms typically "discount the future" in their decision making, but the extent to which they do so varies across species, sexes, age classes, and circumstances. This variability has been studied by biologists, economists, psychologists, and criminologists. We argue that the conceptual framework required for an interdisciplinary synthesis of knowledge in this area is the evolutionary adaptationist analysis of reproductive effort scheduling pioneered by George Williams. PMID- 15884737 TI - Maladaptation and natural selection. AB - The transformations George Williams initiated in evolutionary biology seem so blindingly obvious in retrospect that they spur the question of why he saw what no one else did. While most humans are prone to see only what theory predicts, Williams sees in bold relief whatever does not fit. Not an adaptationist or an anti-adaptationist, Williams is better described as a maladaptionist. The challenge of finding evolutionary explanations for apparent maladaptations has been overlooked with casualness akin to that once typical for group selection. Suboptimal traits tend to be dismissed as illustrations of the weakness and stochastic nature of selection compared with mutation and drift. A closer look suggests that such constraints are only one of six possible kinds of explanations for apparently suboptimal designs: mismatch, coevolution, tradeoffs, constraints, reproductive advantage at the expense of the individual, and defenses that are aversive but useful Medicine has asked proximate questions at every possible level but has only begun to ask evolutionary questions about why bodies are vulnerable to disease. Considering all six possible evolutionary reasons for apparently suboptimal traits will spur progress not only in medicine but also more generally in biology. 'Williams Vision" may not yield a net benefit to the possessor, but it is invaluable for the species. PMID- 15884738 TI - George Williams, theoretician and guerilla environmentalist. AB - George Williams is rightly honored for his contributions to basic biological theory. In addition, however, his thought and contribution paved the way for much needed integration of basic evolutionary theory and modern environmental problems. Specifically, his contributions to the levels of selection" debate, and his application of these contributions to the "Gaia" approach to ecological problems, may help us improve our ability to move past untested prescriptions to a thoughtful matching of the characteristics of the problem and solution, and thus improve our effectiveness. PMID- 15884739 TI - Relationship between diagnosis and disposition of patients admitted to a state psychiatric hospital. AB - The lifetime outcome for individuals diagnosed with affective disorders is generally more favorable than for those diagnosed with a schizophrenic disorder. We determined if a similar differential outcome could be detected among 139 patients hospitalized on the admissions unit of a state psychiatric facility between 1998 and 2001, and diagnosed with a Schizophrenic, Schizoaffective or Affective Disorder. The placement of each patient on discharge was categorized as an outpatient environment, a minimum-security treatment unit, a locked ward, or a highly secure forensic facility. Patients with an affective disorder were significantly less likely than the other two groups to have a co-occurring diagnosis of substance abuse, and they performed better on the neuropsychological assessments. However, the groups did not differ in their discharge placements, or in their length of stay. These findings suggest that resolution of more acute symptomatology may be unrelated to factors associated with long-term outcome for individuals suffering from severe and persistent mental illness. PMID- 15884740 TI - Acute service delivery in a police-mental health program for children exposed to violence and trauma. AB - The Child Development Community Policing Program represents a national model of community-based collaboration between police and mental health professionals for violence-exposed and traumatized children. Administrative data from clinical records of a 24-hour consultation service were examined through stepwise multivariate logistic regression to identify child and event characteristics associated with a direct, in-person response at the time of police contact. Of 2361 children, 809 (34.3%) received a direct, in-person response. Relative to Caucasian children, Hispanic youth were more likely to receive this form of response (OR = 1.36). An acute clinical response was more likely for incidents of gang involvement (OR = 8.12), accidents (OR = 5.21), felony assaults (OR = 2.97), property crimes (OR = 2.30), family violence (OR = 1.53) and psychiatric crises (OR = 1.29). Acute response was less likely when juvenile conduct problems (OR = 0.61), fires (OR = 0.59), child maltreatment (OR = 0.57), and domestic violence (OR = 0.44) were involved. Incidents that were more severe or involved a primary mental health component were related to utilization of intensive CDCP resources. PMID- 15884741 TI - Social isolation, impulsivity and depression as predictors of aggression in a psychiatric inpatient population. AB - Aggressive behavior among psychiatric inpatients remains an issue of concern for staff, families and patients themselves. At the present time, studies examining prediction of aggression among psychiatric inpatients have focused mainly on diagnostic or demographic risk factors. Unfortunately little is known about specific social functioning and personality risk factors that may help identify specific individuals at risk for aggressive behavior. Given that many individuals who have engaged in violent criminal behavior have been observed to experience a combination of social isolation, depression and impulsiveness, it is possible that this same combination of traits may function as a predictor of aggression among psychiatric inpatients. The current study examines whether psychiatric inpatients with a combination of social isolation, depression and impulsivity are significantly more likely to become aggressive than other psychiatric inpatients without that combination of factors. Results indicated that impulsivity functioned as a positive predictor of aggression, whereas depression acted as a protective factor. Perceived social support did not appear to relate strongly to aggression. Further, physicians' ratings of hostility were more predictive of aggressive incidents than were self-reports of hostility. Clinical implications of these findings are discussed. PMID- 15884743 TI - Precipitants of elderly psychiatric patient assaults on staff: preliminary empirical inquiry. AB - Although there have been several studies of the characteristics of psychiatric patient assailants, there have been only six comprehensive, empirical assessments of precipitants to these assaults and no precipitant study has focused solely on elderly psychiatric patient assailants. This one and one-half year, retrospective study continued the inquiry into the nature of patient assault precipitants and focused only on elderly assailants. Older, male patients with a diagnosis of schizophrenia and histories of violence toward others and substance use disorder physically assaulted primarily male, mental health workers. These staff victims experienced disruptions in the domains of mastery, attachment, and meaning as well as the symptomatology associated with psychological trauma. The most common precipitants to these assaults were denial of services and acute psychosis. The findings and implications for health care providers in long-term care settings where elderly psychiatric patients reside are discussed. PMID- 15884742 TI - Psychiatric emergency services: a review of the literature and a proposed research agenda. AB - To gain a better understanding of the scope of previous research on psychiatric emergency services (PES) and to identify areas of focus for future studies, the author reviews PES literature published between 1983 and 2003. PES literature was identified by performing a MEDLINE search. The author was interested in studies or reviews of all types of PES. The refereed research, consisting of 85 articles, is a representation of the existing literature as multiple studies were found to be similar in topic and methodology. This review is organized using Donabedian's structure, process, outcome framework to describe research that has examined 1) providers, clients, and services; 2) variations in access, utilization, evaluation, treatment, and continuity of care; and 3) outcomes of these services. The resulting report reveals methodological limitations that are pervasive throughout the PES literature, dimensions of psychiatric emergency services that remain unexplored, and a PES research agenda. PMID- 15884744 TI - Against the grain? A reasoned argument for not closing a state hospital. AB - In the face of the Massachusetts Governor's attempts to close one of the state's four remaining state hospitals, Massachusetts legislators overrode the Governor's veto of funding for the hospital, but required the state's Mental Health Authority to author a study of the implications of further loss of public sector inpatient beds. The Center for Mental Health Services Research of the University of Massachusetts Medical School conducted its own study concluding that maintaining a longer-term inpatient capacity in the public sector in central Massachusetts was both necessary and accrued a significant number of benefits. This article can serve as a model for the reasoned position that a state hospital in 21st century psychiatry can be looked at as a multiservice center that fulfills a key role in a public sector, integrated system of treatment, care, training and research. PMID- 15884745 TI - Determinants of geropsychiatric inpatient length of stay. AB - Despite efforts to decrease lengths of acute psychiatric hospital stays, some geriatric inpatients continue to have extended stays. This research examined factors related to length of stay (LOS), including legal and administrative factors not traditionally included in prior studies. The charts of 384 patients, representing all 464 discharges from an inpatient geropsychiatric unit over a one year period, were evaluated retrospectively and analyzed using logistic regression and logarithmic transformation. The LOS of over 12% of the inpatients was 26 days or more (average LOS 14.1). Factors significantly associated with longer LOS were: receiving electroconvulsive therapy (ECT), higher Brief Psychiatric Rating Scale (BPRS) positive symptoms scores, falling, pharmacology complications, multiple prior psychiatric hospitalizations, requiring court proceedings to continue hospitalization or medicate against will, consultation delays and not performing ECT on weekends. Neither demographics nor diagnoses alone had influence on length of stay. Incorporation of LOS predictors into Medicare Inpatient Prospective Payment System (IPPS) would more accurately account for the complexity in the cost of caring for geropsychiatry patients. PMID- 15884746 TI - Describing the epidemiology and aetiology of bacterial pneumonia in children: an unresolved problem. PMID- 15884747 TI - Aetiology and clinical presentation of pneumonia in hospitalized and outpatient children in Northeast Brazil and risk factors for severity. AB - Data on presentation, aetiology, and prognostic indicators of childhood pneumonia, which can help design strategies for controlling the disease, are generally scarce in developing countries. In this paper, the distribution of aetiologic agents, clinical presentation, and evolution of pneumonia cases are described, and the factors associated with duration of pneumonia episode and of hospital admission examined. During June 1994-June 1995, 472 children, aged 6-59 months, with clinical diagnosis of pneumonia, who were admitted to hospital or treated as outpatients, were investigated in Recife, Northeast Brazil. Pneumonia, in most cases, was confirmed by radiology. A combination of methods was used for investigating the aetiology of pneumonia. Data obtained on a large number of clinical, socioeconomic and biological variables were analyzed to determine the prognostic factors for the severity and outcome of pneumonia. Bacteria were identified in 26.7% of the cases, while viruses and mixed infections accounted for 8.4% and 2.7% respectively. Haemophilus influenzae (18.9%), Streptococcus pneumoniae (6.4%), and respiratory syncytial virus (5.0%) were most often identified. The pneumonia case-fatality rate was 0.8%. The best clinical predictors of severity were: lung complications at baseline, tachypnoea (for duration of episode), and chest indrawing (for duration of hospital admission). Young age, low birth-weight, and prolonged fever prior to admission to the study also predicted a more prolonged illness, and under-nutrition was a predictor of longer hospital stay. While the development of new vaccines is an important measure for reducing morbidity and mortality due to pneumonia, emphasis on appropriate case management needs to be maintained, with particular attention to children who show the identified risk factors for a poor prognosis. PMID- 15884748 TI - Neonatal mortality in rural Bangladesh: an exploratory study. AB - Bangladesh has a neonatal death rate that is substantially high and demands urgent attention. To assess the causes of neonatal mortality, 1,019 pregnant women were followed up in eight randomly-selected rural areas of the country. Trained female interviewers visited the households of the subjects at four-week intervals to record neonatal deaths (within 28 days after birth). For each death, they administered a structured verbal autopsy questionnaire to the mother and/or a close family member. Based on these field data, three neonatologists arrived at a consensus to assign two causes of death--an originating cause and a direct cause. The neonatal mortality rate was 53.5 per 1,000 livebirths. The originating causes of death were pre-maturity/low birth-weight (30%), difficult labour (16%), unhygienic birth practices (16%), others (4%), and unknown (34%). The direct causes were sepsis (32%), asphyxia (26%), tetanus (15%), respiratory distress (6%), others (6%), and unknown (14%). According to the prevailing causes of neonatal deaths, implementation of intervention programmes, often in the community, that do not depend on highly-technical training or sophisticated equipment should be implemented. PMID- 15884749 TI - Introducing hepatitis B virus vaccine into the Expanded Programme on Immunization in Bangladesh: a proposed method to evaluate whether the existing infrastructure has the capacity. AB - To determine whether the existing Expanded Programme on Immunization (EPI) in Bangladesh has the capacity to introduce the hepatitis B virus (HBV) vaccine, this study was carried out in all the nine health facilities, which maintain a cold-chain, in Chandpur district of Bangladesh. The research, focusing specifically on cold-chain equipment, aimed at developing and applying an indicator of the use of cold-chain equipment. A structured questionnaire, developed and field-tested, was used for collecting information on cold-chain equipment and their use-rate. Data were used for estimating the resources needed to introduce the HBV vaccine and for increasing the coverage of measles and DPT vaccines. The findings of the study showed that the use-rate of cold-chain equipment in this district was low, suggesting that the district has sufficient spare capacity to introduce and sustain the storage of an increased quantity of vaccines. This paper suggests an approach to study capacity in relation to infrastructural facilities. By measuring the capacity of capital equipment, the study has illustrated that the measurement of resource-use rates provides useful information about the burden that a new vaccine places on the EPI. PMID- 15884750 TI - Relationship among fatness, blood lipids, and insulin resistance in Pakistani children. AB - Observations on associations between fatness and metabolic risks among South-East Asian adults have resulted in devising lower thresholds of body mass index (BMI) for them. Metabolic abnormalities, including type 2 diabetes, are now also appearing in children and are associated with obesity. There has not been much work done to identify indicators of metabolic risks among South Asian children. This study was undertaken to observe the relationship among fatness, blood lipids, and insulin resistance in Pakistani children. Fatness, lipids, and insulin resistance were assessed in 92 middle-class Pakistani school children aged 8-10 years. Height, weight, waist, hips, mid-arm circumference, and triceps skin-fold, measured in school, were used for calculating various indicators of fatness, i.e. BMI, waist hip ratio (WHR), and arm-fat percentage. Fasting blood samples were analyzed for total lipids, triglycerides (TG), total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), glucose and insulin levels. Homeostasis model assessment (HOMA) index was calculated to assess insulin resistance. Two separate multiple regression models of various risk indicators (family history, sex, BMI, WHR, arm-fat percentage) showed that only arm-fat percentage had a significant positive association both with insulin levels (b = 2.04, p = 0.044) and LDL (b = 2.11, p = 0.037). Only five children were overweight (BMI-for-age > 85th percentile according to National Center for Health Statistics 2000 reference). Neither overweight children nor those who were in the uppermost tercile of BMI-for-age differed significantly from other children in terms of presence of higher-than-desirable values of lipids or insulin. However, compared to those in the lowest tercile, children who were in the uppermost tercile of armfat percentage had a significantly higher frequency of high blood cholesterol (40% vs 67%, p = 0.027), high LDL (33.3% vs 61.3%, p = 0.026), and markedly higher proportion above average insulin levels (16.7% vs 35.5%, p = 0.083). Arm-fat percentage could be developed as a practical tool for determining the risk status of children. However, further cross-sectional assessments are needed to ascertain accurate relationships among arm-fat percentage, lipid profiles, and insulin resistance in larger and varied groups of children. PMID- 15884751 TI - Diversity of Vibrio cholerae strains isolated in Delhi, India, during 1992-2000. AB - The National Institute of Communicable Diseases (NICD), Delhi, India, conducts a laboratory-based surveillance of cholera cases referred from the Infectious Disease Hospital, Delhi. The prevalence and antimicrobial susceptibilities of Vibrio cholerae O1, O139, and others, isolated from cholera patients for nine years, were analyzed to determine the changing trends in their isolation and drug resistance patterns. In total, 29,196 stool samples or rectal swabs, collected during April 1992-December 2000, were included in this study. Of these, 13,730 (47%) were positive for V. cholerae: 11,091 for V. cholerae O1 (80.7%) and 1,943 (14%) for V. cholerae O139, and 696 (5%) were non-O1 and non-O139. V. cholerae O1 was the dominant serotype during 1992-1993, when V. cholerae O139 emerged as a new serotype but, thereafter, remained low from 1994 to 1999. Phenotypically, re emerged V. cholerae O139 in 2000 displayed a difference compared to those that appeared in 1992-1993, in that the current O139 strains were sensitive to co trimoxazole. Resistance to nalidixic acid and furazolidone was constantly high (100%) among strains of V. cholerae O1 and O139. All strains of V. cholerae were uniformly susceptible to chloramphenicol, tetracycline, amikacin, and norfloxacin. Molecular studies revealed different clones of V. cholerae O1 and O139 prevailing in the country with the re-emergence of V. cholerae O139 of a different clonality in Delhi in 2000, which is likely to play a critical role in temporal antigenic variation among the serogroups of V. cholerae. PMID- 15884752 TI - Low osmolar oral rehydration salts solution in the treatment of acute watery diarrhoea in neonates and young infants: a randomized, controlled clinical trial. AB - To compare the efficacy and safety of low osmolar oral rehydration salts solution (ORS-75) (mmol/L: Na+ 75, osmolarity 245) with that of World Health Organization recommended ORS (ORS-90) (mmol/L: Na+ 90, osmolarity 311 ) in the treatment of acute watery diarrhoea in neonates and very young infants, a randomized double blind, controlled clinical trial was carried out at the Clinical Research and Service Centre of ICDDR,B: Centre for Health and Population Research, Dhaka, Bangladesh, during January 1998-December 1999. Infants, aged < or = 2 months, presenting with a history of watery diarrhoea of < or = 72 hours, with no or some dehydration and without any systemic illness, were randomly assigned to receive either ORS-75 or ORS-90 for the correction and subsequent prevention of dehydration. Infants were studied for a maximum of five days. Total stool output, stool frequency, and requirement for ORS were outcome measures. Serum electrolytes were measured at 24 hours after admission to monitor serum sodium imbalance. Seventy-three infants received ORS-75, and 71 received ORS-90. Both the groups were comparable in their baseline characteristics. Diarrhoea resolved within five days in 53% and 66% of infants receiving ORS-75 and ORS-90 respectively (p = 0.3). Total stool volume [median (inter-quartile range) 132 (65 280) vs 139 (70-259) g/kg, p = 0.9], during the study period, was not significantly different between the two groups. Total stool frequency [31 (16-51) vs 35 (16-53), p = 0.9] and total ORS intake [192 (96-374) vs 209 (134-317) mL/kg, p = 0.7] were similar between the groups. No infants developed late evidence of hypernatraemia, irrespective of treatment. The results of the study indicate that ORS-75 is as safe as standard ORS-90 in the treatment of acute watery diarrhoea in neonates and very young infants and is effective in correcting and preventing dehydration. PMID- 15884753 TI - HIV counselling and testing of pregnant women attending antenatal clinics in Botswana, 2001. AB - This study examined the extent to which women accessing antenatal-care services in Botswana were offered HIV-related information and counselling and an opportunity to take an HIV test as part of the Prevention of Mother-to-Child Transmission of infection, and how these women responded. Data for this study were drawn from the Botswana AIDS Impact Survey 2001, a nationally-representative sample survey. The survey successfully interviewed over 4,494 of 4,728 eligible women on various issues relating to HIV/AIDS at both household and individual levels. Frequencies, cross tabulations, and logistic regression were used for data analysis. Over half (57.9%) of the women were offered HIV/AIDS-related information, counselling, or testing. Age, education, and residence were important predictors of being offered HIV counselling or testing. Younger and more-educated women and those residing in towns were more likely to be offered both HIV counselling and testing than older, less-educated, and rural women. Seventy-nine percent of the women who were offered HIV testing agreed to undergo the test regardless of their background characteristics. The number of pregnant women who underwent HIV testing during antenatal care accounted for only a fifth (21%) of all antenatal-care attendees in 2001. Lack of capacity to deliver voluntary counselling and testing services to all pregnant women attending antenatal care is one of the biggest challenges to increased use of voluntary counselling and testing services. PMID- 15884754 TI - Intra-class correlation estimates for assessment of vitamin A intake in children. AB - In many community-based surveys, multi-level sampling is inherent in the design. In the design of these studies, especially to calculate the appropriate sample size, investigators need good estimates of intra-class correlation coefficient (ICC), along with the cluster size, to adjust for variation inflation due to clustering at each level. The present study used data on the assessment of clinical vitamin A deficiency and intake of vitamin A-rich food in children in a district in India. For the survey, 16 households were sampled from 200 villages nested within eight randomly-selected blocks of the district. ICCs and components of variances were estimated from a three-level hierarchical random effects analysis of variance model. Estimates of ICCs and variance components were obtained at village and block levels. Between-cluster variation was evident at each level of clustering. In these estimates, ICCs were inversely related to cluster size, but the design effect could be substantial for large clusters. At the block level, most ICC estimates were below 0.07. At the village level, many ICC estimates ranged from 0.014 to 0.45. These estimates may provide useful information for the design of epidemiological studies in which the sampled (or allocated) units range in size from households to large administrative zones. PMID- 15884755 TI - Perceptions of tuberculosis: attributions of cause, suggested means of risk reduction, and preferred treatment in the Limpopo province, South Africa. AB - Despite the availability of effective treatment regimens, exceptionally high rates of tuberculosis persist in South Africa. A better understanding of public perceptions of tuberculosis is important for the improvement of the existing tuberculosis-control programmes. This study was aimed at investigating public perceptions about causes, suggested means of risk reduction, and preferred treatment of tuberculosis. The cross-sectional study included a sample of 80 community members from four different age groups chosen by quota sampling in a semi-urban community. Results indicated that 13.75% of the community members reported biomedical causes of tuberculosis, 35% suggested biomedical methods as a means for risk reduction, and 91.25% preferred medical treatment. Significant gender but not age difference was found. Women reported more often than men a biomedical cause of tuberculosis. A number of misconceptions about the cause and risk reduction of tuberculosis were also mentioned. The results suggest that people in a semi-urban area of the Limpopo province have limited knowledge and understanding of tuberculosis. It is suggested that healthcare workers provide services for increasing basic knowledge about causes and risk reduction of tuberculosis of the population in every age group. PMID- 15884756 TI - Marital disruption: determinants and consequences on the lives of women in a rural area of Bangladesh. AB - This study, carried out during the second half of 1995, investigated the predisposing factors leading to marital disruption and its consequences on the lives of women in Matlab, a rural area of Bangladesh. Both qualitative and quantitative methods were used. Data were generated from detailed case studies and quantitative surveys of a small number of maritally-disrupted women. Additional data were used from the ongoing demographic surveillance system of ICDDR,B: Centre for Health and Population Research. The findings revealed that divorced and abandoned women and their children were extremely vulnerable, both socially and economically. Various factors that influence marital disruption were identified, the most important ones being: aspects determining the process of marriage, various family problems due to non-fulfillment of demand for dowry, mutual distrust, extramarital relationships, quality of sexual life, education of women, and other behavioural characteristics of individuals. Level of education of the wife showed an inverse relationship with the risk of divorce. Women who did not have livebirths from their first pregnancy had a higher risk of divorce. The effect of pregnancy outcome was dependent on the level of education of women. Illiterate women with unsuccessful pregnancy outcomes were at the highest risk of being divorced, with the lowest risk for women with some education and a livebirth. The findings clearly indicate the need for broad-based social development programmes for women, especially to enhance their education to reduce their vulnerability to marital instability and its consequences. PMID- 15884757 TI - Intimate partner violence against women: experiences from a woman-focused development programme in Matlab, Bangladesh. AB - This paper explores the association between microcredit-based development programmes and domestic violence against women perpetrated by their husbands. A sub-set of cross-sectional data collected in 1999 from 60 BRAC-ICDDR,B study villages in Matlab, Bangladesh, was used. Data were analyzed to characterize group-level differences among study women regarding the reported occurrence of violence (physical and/or mental) and to identify its predictors. About 17.5% of women had experienced violence from their husbands in the past four months, the proportion being greater among BRAC households (p = 0.05). Results of logistic regression identified age, schooling, age of household head, and self-rated poverty status of household as important predictors of violence, but not level of BRAC membership. The study concludes that the greater level of domestic violence reported during the initial stages of BRAC membership subsided with the introduction of skill-development training among participant women over time. PMID- 15884758 TI - Coffee phagia and iron-deficiency anaemia: a possible association with Helicobacter pylori. PMID- 15884759 TI - n-3 fatty acid enrichment of edible tissue of poultry: a review. AB - There is clear evidence of the nutritional benefits of consuming long-chain n-3 PUFA, which are found predominantly in oily fish. However, oily fish consumption, particularly in the United Kingdom, is declining, as is the consumption of all meats with the exception of poultry, which has increased in consumption by 73% in the last 30 yr. This pattern, if less marked, is reflected throughout Europe, and therefore one means of increasing long-chain n-3 PUFA consumption would be to increase the long-chain n-3 PUFA content in the edible tissues of poultry. This review considers the feasibility of doing this, concentrating particularly on chickens and turkeys. It begins by summarizing the benefits to human health of consuming greater quantities of n-3 FA and the sources of n-3 PUFA in the human diet. The literature on altering the FA composition of poultry meat is then reviewed, and the factors affecting the incorporation of n-3 PUFA into edible tissues of poultry are investigated. The concentration of alpha-linolenic acid (ALA) in the edible tissues of poultry is readily increased by increasing the concentration of ALA in the birds' diet (particularly meat with skin, and dark meat to a greater extent than white meat). The concentration of EPA in both white and dark meat is also increased when the birds' diet is supplemented with EPA, although supplementing the diet with the precursor (ALA) does not result in a noticeable increase in EPA content in the edible tissues. Although supplementing the birds' diets with relatively high concentrations of DHA does result in an increased concentration of DHA in the tissues, the relationship between dietary and tissue concentrations of DHA is much weaker than that observed with ALA and EPA. The impact that altering the FA composition of edible poultry tissue may have on the organoleptic and storage qualities of poultry products is also considered. PMID- 15884760 TI - Maternal dietary conjugated linoleic acid alters hepatic triacylglycerol and tissue fatty acids in hatched chicks. AB - The effects of feeding CLA to hens on newly hatched chick hepatic and carcass lipid content, liver TAG accumulation, and FA incorporation in chick tissues such as liver, heart, brain, and adipose were studied. These tissues were selected owing to their respective roles in lipid assimilation (liver), as a major oxidation site (heart), as a site enriched with long-chain polyunsaturates for function (brain), and as a storage depot (adipose). Eggs with no, low, or high levels of CLA were produced by feeding hens a corn-soybean meal-basal diet containing 3% (w/w) corn oil (Control), 2.5% corn oil + 0.5% CLA oil (CLA1), or 2% corn oil + 1.0% CLA oil (CLA2). The egg yolk content of total CLA was 0.0, 1.0, and 2.6% for Control, CLA1, and CLA2, respectively (P< 0.05). Maternal dietary CLA resulted in a decrease in chick carcass total fat (P < 0.05). Liver tissue of CLA2 chicks had the lowest fat content (P < 0.05). The liver TAG content was 8.2, 5.8, and 5.1 mg/g for Control, CLA1, and CLA2 chicks, respectively(P < 0.05). The chicks hatched from CLA1 and CLA2 incorporated higher levels of cis-9,trans-11 CLA in the liver, plasma, adipose, and brain than Control (P< 0.05). The content of 18:0 was higher in the liver, plasma, adipose, and brain of CLA1 and CLA2 than Control (P< 0.05), but no difference was observed in the 18:0 content of heart tissue. A significant reduction in 18:1 was observed in the liver, plasma, adipose, heart, and brain of CLA1 and CLA2 chicks (P< 0.05). DHA (22:6n-3) was reduced in the heart and brain of CLA1 and CLA2 chicks (P < 0.05). No difference was observed in carcass weight, dry matter, or ash content of chicks (P> 0.05). The hatchabilities of fertile eggs were 78, 34, and 38% for Control, CLA1, and CLA2, respectively (P< 0.05). The early dead chicks were higher in CLA1 and CLA2 than Control (18 and 32% compared with 9% for Control), and alive but not hatched chicks were 15 and 19% for CLAl and CLA2, compared with 8% for Control (P < 0.05). Maternal supplementation with CLA leads to a reduction in hatchability, liver TAG, and carcass total fat in newly hatched chicks. PMID- 15884761 TI - Effects of specific CLA isomers on plasma fatty acid profile and expression of desaturases in humans. AB - Human studies suggest that CLA changes metabolism, possibly through effects on mRNA expression of desaturase and elongase enzymes. In this respect, differential effects of the two most common dietary CLA isomers, cis-9,trans-11 (c9,t11) and trans-10,cis-12 (t10,c12) CLA, have hardly been studied. We therefore gave 25 healthy, overweight men and women daily for 6 wk a drinkable dairy product containing 3 g of oil that was rich in oleic acid. For the next 18 wk, the control group (n = 7) continued to use this product, whereas the second (n = 9) and third groups (n = 9) received products with 3 g of purified c9,t11 CLA or t10,c12 CLA. For each gram of c9,t11 CLA consumed, the proportion in plasma phospholipids increased by 0.26%. For t10, c12 CLA, this value was 0.20%. The t10,c12 CLA isomer increased plasma TAG levels of conjugated 18:3, whereas c9,t11 CLA increased those of both conjugated 18:3 and 20:3. In plasma phospholipids, the delta9 desaturation index of 18:0 (18:1 n-9/18:0) was decreased by t10,c12 CLA (P= 0.03 for diet effects), and the delta6 desaturation index [(18:3n-6 + 20:3n-6)/18:2n-6] was decreased by both CLA isomers (P < 0.01 for diet effects). The delta5 desaturation index (20:4n-6/20:3n-6) and the delta9 desaturation index of 16:0 (16:1 n-7/16:0) were not affected. No effects were seen on mRNA expression of desaturases and elongase in peripheral blood mononuclear cells (PBMC). We therefore conclude that incorporation of c9,t11 and t10,c12 CLA into plasma lipids reflects dietary intakes. Compared with oleic acid, delta9 and delta6 desaturation indices in plasma phospholipids are decreased after consumption of c9,t11 or t10,c12 CLA. Effects on desaturation indices were, however, not reflected by changes at the transcriptional level for the various desaturases and elongase enzymes in PBMC. PMID- 15884762 TI - Synthesis of the conjugated trienes 5E,7E,9E,14Z,17Z-eicosapentaenoic acid and 5Z,7E,9E,14Z,17Z-eicosapentaenoic acid, and their induction of apoptosis in DLD-1 colorectal adenocarcinoma cells. AB - During the course of our recent study on the anti-tumor effect of conjugated eicosapentaenoic acids (CEPA), we found that acid mixtures prepared by treating EPA with KOH in ethylene glycol induced potent apoptotic cell death in human tumor cells via membrane phospholipid peroxidation. Interestingly, the KOH treated CEPA mixtures were more cytotoxic than EPA and CLA and had no effect on normal human fibroblast cells. To identify the specific cytotoxic FA in the CEPA mixture, we synthesized possible candidates for the active species. Here, we report the synthesis of (5E,7E,9E, 14Z, 17Z)-5,7,9,14,1 7-eicosapentaenoic acid (E-CEPA) and its 5-(Z) isomer (Z-CEPA), both of which are conjugated trienes that exist naturally in red algae (Ptilota filicina J. Agardh). E-CEPA and Z-CEPA were synthesized from methyl 5-oxopentanoate in six steps, using three types of Wittig reactions as the key steps. Next, we examined the cytotoxicity of E-CEPA and Z CEPA in human tumor cells and confirmed their bioactivity. Both E-CEPA and Z-CEPA had a strong cytotoxic reaction in tumor cells, and this effect occurred through induction of apoptosis. PMID- 15884763 TI - Production and protein kinase C activation of diacylglycerols containing polymethylene-interrupted PUFA. AB - Sciadonic acid (20:3, delta-5c,11 c,14c) is a polymethylene-interrupted PUFA (PMI PUFA) that is present in conifer seeds and known to be incorporated into animal cells and to accumulate in membrane PI as a substitute for arachidonate. In this study, we investigated whether PI having sciadonate could serve as source of DAG that could activate protein kinase C (PKC). When Swiss 3T3 cells cultured with sciadonic acid were stimulated with 100 nM of bombesin, 1-stearoyl-2-sciadonoyl glycerol (G) and 1-stearoyl-2-arachidonoyl-G were produced. The net increments of these two molecular species of DAG reflected the levels of the two molecular species in the PI in the cells. When cells cultured with juniperonic acid (20:4, delta-5c,11c,14c,17c) were stimulated, 1-stearoyl-2-juniperonoyl-G was produced in proportion to the level of this molecular species in PI in the cells. We also examined PKC activation by synthetic DAG using a partially purified PKC fraction from rat brain and found that both 1-stearoyl-2-sciadonoyl-G and 1-stearoyl-2 juniperonoyl-G could activate PKC comparably to 1 -stearoyl-2-arachidonoyl-G. These results indicate that 1-stearoyl-PI having these C20 PMI-PUFA residues can serve as sources of potential signaling molecules. PMID- 15884764 TI - Decreased serum adiponectin in adolescents and young adults with familial primary hypercholesterolemia. AB - Decreased serum adiponectin is associated with dyslipidemia. However, serum adiponectin status has never before been studied in patients with familial related severe primary hypercholesterolemia (FRSPH). The aim of this study is to measure serum adiponectin level in a group of young patients with FRSPH and determine its correlation with insulin-resistant status. Twenty-three patients with FRSPH [average LDL-cholesterol (LDL-C) = 250.8 (190-610) mg/dL] without clinical manifestations of metabolic syndrome as well as 46 healthy (control) adolescents and young adults (<30 yr old) were included. The serum adiponectin, fasting sugar, insulin, lipids, systolic and diastolic blood pressure (SBP and DBP), and anthropometrical indices such as body mass index and waist circumference were obtained. The homeostasis model assessment (HOMA) was calculated to estimate the insulin resistant status. Compared with healthy controls, patients with FRSPH had a significantly lower mean serum adiponectin level (7.7+/-1.8 microg/mL vs. 10.1+/-4.3 microg/mL, P= 0.013). After adjustment for HOMA and associated covariates, multiple linear regression analysis showed that patients with FRSPH are significantly associated with hypoadiponectinemia. Compared with healthy controls, patients with FRSPH had a significantly lower mean serum adiponectin level (7.7 +/-1.8 microg/mL vs. 10.1+/-4.3 microg/mL, P = 0.013). After adjustment for HOMA and associated covariates, multiple linear regression analysis showed that patients with FRSPH are significantly associated with hypoadiponectinemia. The serum adiponectin levels are lower in young patients with FRSPH without clinical manifestations of metabolic syndrome. The mechanism of hypoadiponectinemia in patients with FRSPH is probably independent of insulin resistance. PMID- 15884765 TI - Effects of a diet high in plant sterols, vegetable proteins, and viscous fibers (dietary portfolio) on circulating sterol levels and red cell fragility in hypercholesterolemic subjects. AB - Plant sterols, soy proteins, viscous fibers, and nuts are advised for cholesterol reduction, but their combined effect on plant sterol absorption has never been tested. We assessed their combined action on serum sterols in hyperlipidemic subjects who were following low-saturated fat diets before starting the study and who returned to these diets post-test. The 1-mon test (combination) diet was high in plant sterols (1 g/1,000 kcal), soy protein (23 g/1,000 kcal), viscous fiber (9 g/1,000 kcal), and almonds (14 g/1000 kcal). Fasting blood was obtained for serum lipids and sterols, and erythrocytes were obtained for fragility prior to and at 2-wk intervals during the study. The combination diet raised serum campesterol concentrations by 50% and beta-sitosterol by 27%, although these changes were not significant after Bonferroni correction; near-maximal rises were found by the end of the first week, but no change was found in red cell fragility despite a 29% reduction in the LDL cholesterol level. No significant associations were observed between changes in red cell fragility and blood lipids or sterols. We conclude that plant sterols had a minimal impact on serum sterol concentrations or red cell fragility in hyperlipidemic subjects on diets that greatly reduced their serum lipids. PMID- 15884766 TI - Effects of a water-soluble phytostanol ester on plasma cholesterol levels and red blood cell fragility in hamsters. AB - The aim of this study was to assess the efficacy of a novel water-soluble phytostanol analog, disodium ascorbyl phytostanyl phosphates (DAPP), on plasma lipid levels and red blood cell fragility in hamsters fed atherogenic diets. For 5 wk, 50 male Golden Syrian hamsters were fed a semipurified diet without added cholesterol (noncholesterol, group 1), or a semipurified diet with 0.25% cholesterol (cholesterol-control, group 2). Groups 3-5 were fed the cholesterol control diet with an addition of 1% phytostanols (diet 3), 0.71% DAPP (DAPP 0.7%, diet 4), or 1.43% DAPP (DAPP 1.4%, diet 5). Diets 4 and 5 provided 0.5 and 1% phytostanols, respectively. Supplementation of 0.71 and 1.43% DAPP decreased plasma total cholesterol concentrations by 34 (P < 0.001) and 46% (P< 0.001), respectively, in comparison with the cholesterol-control group, whereas free stanols reduced (P = 0.007) plasma cholesterol concentrations by 14%. Similarly, non-HDL-cholesterol concentrations were reduced by 39 (P < 0.001) and 54% (P < 0.001) in hamsters supplemented with DAPP 0.7% and DAPP 1.4%, respectively, relative to the cholesterol-control group. The hypocholesterolemic effect of DAPP 1.4% was threefold stronger than that of free stanols. In hamsters supplemented with DAPP 1.4%, plasma TG concentrations were 45% lower (P= 0.018) than in cholesterol-control-fed hamsters, whereas no such beneficial effect was observed in the free stanol group. Erythrocyte fragility was unaffected by DAPP or free phytostanols. Results of the current study demonstrate that DAPP lowers cholesterol more efficiently than free stanols, without an adverse effect on erythrocyte fragility in hamsters. PMID- 15884767 TI - A comparison of the effect of medium- vs. long-chain triglycerides on the in vitro solubilization of cholesterol and/or phytosterol into mixed micelles. AB - Despite clinical evidence of the cholesterol-lowering effects of phytosterols, the exact mechanisms involved are still unclear. Displacement of cholesterol by phytosterols from mixed micelles, which is due to their greater hydrophobicity, is one of the hypotheses for the lumenal effects contributing to the reduction of intestinal cholesterol absorption. In this study a dynamic in vitro lipolysis method was used to examine the solubilization behavior of cholesterol and/or phytosterols during lipolysis to probe the efficacy of cholesterol displacement from mixed micelles by phytosterols. The effects of lipid chain length on sterol solubilization were studied by using microcrystalline suspensions containing 17% phytosterol or cholesterol, formulated in long-chain TG (LCT) and medium-chain TG (MCT). When digesting cholesterol-suspended in LCT, the entire cholesterol dose was incorporated into the micellar phase. For the cholesterol formulation suspended in MCT, 50.3% of the initial dose was recovered in the micelles. Under the respective conditions, we observed lower solubilization of phytosterols than of cholesterol (roughly fourfold). Only 25% of the initial phytosterol dose was solubilized from suspensions formulated with LCT, and 13% was solubilized from MCT formulations. Co-administration of phytosterol and cholesterol suspensions showed a significant reduction of cholesterol solubilization, particularly when dosed in MCT, with approximately 25% of the cholesterol dose solubilized. Insignificant amounts of cholesterol were displaced by phytosterols when cholesterol was presolubilized in the mixed micelles. The results show that, compared with LCT, mixed micelles containing MCT lipolysis products have a reduced solubilizing capacity for cholesterol, which adds to the effectiveness of the phytosterols in displacing cholesterol. This suggests potential benefits of using medium chain length lipids in cholesterol-lowering phytosterol products. PMID- 15884768 TI - A study on the causes for the elevated n-3 fatty acids in cows' milk of alpine origin. AB - The influence of grass-only diets either from rye-grass-dominated lowland pastures (400 m above sea level) or botanically diverse alpine pastures (2000 m) on the FA profile of milk was investigated using three groups of six Brown Swiss cows each. Two groups were fed grass-only on pasture (P) or freshly harvested in barn (B), both for two experimental periods in the lowlands and, consecutively, two periods on the alp. Group C served as the control, receiving a silage concentrate diet and permanently staying in the lowlands. Effects of vegetation stage or pasture vs. barn feeding on milk fat composition were negligible. Compared with the control, alpha-linolenic acid (18:3n-3) consumption was elevated in groups P and B (79%; P< 0.001) during the lowland periods but decreased on the alp to the level of C owing to feed intake depression and lower 18:3n-3 concentration in the alpine forage. Average 18:3n-3 contents of milk fat were higher in groups P and B than in C by 33% (P< 0.01) at low and by 96% (P < 0.001) at high altitude, indicating that 18:3n-3 levels in milk were to some extent independent of 18:3n-3 consumption. The cis-9, trans-11 CLA content in milk of grass-fed cows was higher compared with C but lower for the alpine vs. lowland periods whereas the trans-11, cis-13 isomer further increased with altitude. Long-chain n-3 FA and phytanic acid increased while arachidonic acid decreased with grass-only feeding, but none of them responded to altitude. Grass only feeding increased milk alpha-tocopherol concentration by 86 and 134% at low and high altitude (P < 0.001), respectively. Changes in the ruminal ecosystem due to energy shortage or specific secondary plant metabolites are discussed as possible causes for the high 18:3n-3 concentrations in alpine milk. PMID- 15884769 TI - Quantitative determination of total lipid hydroperoxides by a flow injection analysis system. AB - A flow injection analysis (FIA) system coupled with a fluorescence detection system using diphenyl-1-pyrenylphosphine (DPPP) was developed as a highly sensitive and reproducible quantitative method of total lipid hydroperoxide analysis. Fluorescence analysis of DPPP oxide generated by the reaction of lipid hydroperoxides with DPPP enabled a quantitative determination of the total amount of lipid hydroperoxides. Use of 1-myristoyl-2-(12-((7-nitro-2-1,3-benzoxadiazol-4 yl)amino) dodecanoyl)-sn-glycero-3-phosphocholine as the internal standard improved the sensitivity and reproducibility of the analysis. Several commercially available edible oils, including soybean oil, rape-seed oil, olive oil, corn oil, canola oil, safflower oil, mixed vegetable oils, cod liver oil, and sardine oil were analyzed by the FIA system for the quantitative determination of total lipid hydroperoxides. The minimal amounts of sample oils required were 50 microg of soybean oil (PV = 2.71 meq/kg) and 3 mg of sardine oil (PV = 0.38 meq/kg) for a single injection. Thus, sensitivity was sufficient for the detection of a small amount and/or low concentration of hydroperoxides in common edible oils. The recovery of sample oils for the FIA system ranged between 87.2+/-2.6% and 102+/-5.1% when PV ranged between 0.38 and 58.8 meq/kg. The CV in the analyses of soybean oil (PV = 3.25 meq/kg), cod liver oil (PV = 6.71 meq/kg), rapeseed oil (PV = 12.3 meq/kg), and sardine oil (PV = 63.8 meq/kg) were 4.31, 5.66, 8.27, and 11.2%, respectively, demonstrating sufficient reproducibility of the FIA system for the determination of lipid hydroperoxides. The squared correlation (r2) between the FIA system and the official AOCS iodometric titration method in a linear regression analysis was estimated at 0.9976 within the range of 0.35-77.8 meq/kg of PV (n = 42). Thus, the FIA system provided satisfactory detection limits, recovery, and reproducibility. The FIA system was further applied to evaluate changes in the total amounts of lipid hydroperoxides in fish muscle stored on ice. PMID- 15884770 TI - sn-position determination of phospholipid-linked fatty acids derived from erythrocytes by liquid chromatography electrospray ionization ion-trap mass spectrometry. AB - The sn-position of FA in membrane lipids has an influence on the physiological function of cells, is predictive for diseases, and therefore is useful for diagnostics. The current study compares the compositions of acyl chain substituents in the sn-1 and sn-2 positions of the glycerol backbones of phospholipids derived from human erythrocytes by using RP-HPLC coupled with on line electrospray ionization ion trap MS. Preferential loss of the acyl group in the sn-1 position was used to determine the degree of regiospecific preference exhibited by the phospholipid molecules. The identities of the molecular species and the positions of the acyl substituents were identified using product-ion spectra of major precursor ions selected from the mass spectra averaged across peaks in the total ion chromatogram. Saturated FA were found to be located mainly in the sn-1 position of the glycerol backbones of erythrocyte phospholipids, whereas PUFA were found primarily in the sn-2 position. All measured phospholipids revealed palmitic acid (16:0) at the sn-1 position. Linoleic acid (18:2n-6) and arachidonic acid (20:4n-6) were found to be attached exclusively to the sn-2 position of the backbone, whereas eicosadienoic (20:2n-6) and eicosatrienoic acid (20:3n-9) occurred in both positions of the backbone of PC. Oleic (18:1n-9), linoleic (18:2n-6), and octadecatrienoic (18:3) acids of PE and PS were linked to both positions. Lignoceric acid (24:1 n-9) was found to be strictly localized at the sn-2 position, whereas nervonic (24:1n-9) acid of PS was associated with both positions of the backbone. A detailed analysis of the blood cell membrane lipids by MS might be helpful to characterize postprandial kinetics of pharmacological or dietary lipid applications, as well as environmental influences on cell membranes. PMID- 15884771 TI - Rodent models of rheumatoid arthritis. AB - The study of rheumatoid arthritis is greatly facilitated by animal models that enable investigation of a complex system involving inflammation, immunological tolerance, and autoimmunity. Although the models cover several species and pathogenetic mechanisms and can be classified as induced or spontaneous, all converge on arthritis. However, because each model features a different mechanism driving disease expression, the merits of each should be evaluated carefully in making the appropriate choice for the scientific question to be addressed. In addition, because the incidence and kinetics of disease vary by model, careful thought should be given to protocol design to minimize animal use. PMID- 15884772 TI - Low-dose infectivity of Staphylococcus aureus (SMH strain) in traumatized rat tibiae provides a model for studying early events in contaminated bone injuries. AB - Animal models of post-traumatic acute osteomyelitis (OM) that closely mimic human scenarios, including infection prophylactic procedures such as debridement and lavage, may provide a better understanding of OM. We contaminated mechanically traumatized rat tibiae (n = 69) with various doses of a Staphylococcus aureus strain (SMH) known to cause human OM and then performed curettage and lavage. Tibiae were harvested 24 h after lavage for assessment of bacterial load and determination of minimal infective doses for 50% (ID50) and 95% (ID95) of rats. Some experiments varied tibial harvest time after lavage (n = 10); for progressive infection, tibiae were evaluated at 7 and 15 days after contamination (n = 17 for each time point). At 24 h after contamination, the ID50 was 1.8 x 10(3) CFU, and the ID95 was 9.2 x 10(3) CFU. Tibial bacterial loads did not increase with inocula greater than the ID95. Lavage removed many bacteria from bone, but it did not prevent subsequent infection or disease. At 15 days after contamination, most tibiae (14 of 17) were infected, with macroscopic and radiological signs of established OM. This newly described rat OM model, with a low ID95 despite prophylactic curettage and lavage, closely mimics events in contaminated human bone injuries. This situation will allow study of early factors in contaminated bone injuries, including clinical interventions that may reduce infection and prevent disease. PMID- 15884773 TI - Birth of rhesus macaque (Macaca mulatta) infants after in vitro fertilization and gestation in female rhesus or pigtailed (Macaca nemestrina) macaques. AB - A study was conducted to assess the possibility of using pigtailed macaques (Macaca nemestrina) as recipients for rhesus macaque (Macaca mulatta) embryos. A total of 250 oocytes were collected from 11 rhesus monkeys during 12 follicular aspirations. We performed 15 embryo transfers with two embryos each into rhesus recipients, which resulted in eight pregnancies, of which two were lost during the second trimester. Among the remaining six pregnant rhesus macaques, two were carrying twins, resulting in the birth of eight infants. Twelve transfers of rhesus embryos into pigtailed macaques resulted in one pregnancy and the birth of one infant. Fetal growth and development were monitored by monthly ultrasound examinations, during which biparietal measurements were taken and compared with those derived from 22 pregnant control monkeys. In vitro fertilization-derived singletons tended to develop faster than did twins and naturally conceived control singletons during the initial months of pregnancy and weighed more at birth than did twins. There were pronounced morphologic changes in the placenta of the rhesus that developed in the female pigtailed macaque. These included an irregular shape, elevated placenta-to-birth-weight ratio, and an abnormal length and diameter of the umbilical cord. Histologic analyses of the rhesus-pigtailed placenta showed evidence of maternal-placental floor infarction and thrombosis of the spiral artery with resulting infarction of the villi. These results demonstrate that pigtailed macaques can carry rhesus fetuses to term, but further studies are necessary to determine the cause of the decreased pregnancy rates and observed placental abnormalities. PMID- 15884774 TI - Use of frozen-thawed oocytes for efficient production of normal offspring from cryopreserved mouse spermatozoa showing low fertility. AB - Freezing of spermatozoa and unfertilized oocytes is a useful tool for the conservation of mouse genetic resources. However, the proportion of frozen-thawed oocytes fertilized with spermatozoa in vitro is low because spermatozoa, especially those frozen-thawed, can not penetrate into oocytes because of hardening of the zona pellucida following premature release of cortical granules. To produce offspring efficiently from cryopreserved transgenic mouse gametes, we fertilized frozen-thawed gametes by using intracytoplasmic sperm injection (ICSI) and assessed pre- and postimplantation development of embryos. Compared with fresh unfertilized oocytes, frozen-thawed unfertilized oocytes were highly tolerant to damage by injection, as the survival rates after injection of frozen spermatozoa were 51 and 78%, respectively. Frozen-thawed oocytes that survived after sperm injection developed normally to the blastocyst stage and gave rise to offspring. Moreover, offspring with transgenes also were obtained from frozen gametes fertilized by ICSI. These results demonstrate that ICSI is an efficient technique for producing offspring from transgenic spermatozoa showing low fertility and that use of frozen-thawed oocytes leads to conservation of genetic resources because suboptimally preserved gametes are not wasted. PMID- 15884775 TI - Relation between storage temperature and fertilizing ability of freeze-dried mouse spermatozoa. AB - The advantage of freeze-dried mouse spermatozoa is that samples can be stored in the refrigerator (+4 degrees C). Moreover, the storage of freeze-dried spermatozoa at ambient temperature would permit spermatozoa to be shipped easily and at low cost around the world. To examine the influence of the storage temperature on freeze-dried spermatozoa, we assessed the fertilizing ability of spermatozoa stored at different temperatures. Cauda epididymal spermatozoa were freeze-dried in buffer consisting of 50 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, 50 mM NaCl, and 10 mM Tris-HCl (pH 8.0). Samples of freeze-dried spermatozoa were stored at -70, -20, +4, or +24 degrees C for periods of 1 week and 1, 3, and 5 months. Sperm chromosomes were maintained well at -70, -20, and + 4 degrees C for 5 months, and oocytes fertilized with these spermatozoa developed to normal offspring. Moreover, the chromosomal integrity of spermatozoa stored at -20 or + 4 degrees C did not decrease even after 17 months. In contrast, the chromosomes of spermatozoa stored at +24 degrees C were maintained well for 1 month but became considerably degraded after 3 months. In addition, to investigate the cause of deterioration of sperm chromosomes during storage at +24 degrees C, spermatozoa were freeze-dried in buffer containing DNase I. The chromosomes of spermatozoa freeze-dried with 1 or 0.2 units/ml of DNase I, 100% or 72%, respectively, exhibited chromosomal abnormalities. Our findings suggest that freeze-dried spermatozoa can be stored long-term with stability at +4 degrees C, and the suppression of nucleases present in the buffer or spermatozoa during storage led to the achievement of long-term storage of freeze-dried spermatozoa. PMID- 15884776 TI - Survey of captive cynomolgus macaque colonies for SRV/D infection using polymerase chain reaction assays. AB - The exogenous simian type D retroviruses (SRV/Ds) are prevalent in macaque monkeys and sometimes cause immunodeficiency with anemia, weight loss, and persistent unresponsive diarrhea. SRV/D isolates are classified as subtypes 1 to 6, and the entire sequences of the gag region of SRV/D-1, -2, and -3 and SRV/D Tsukuba (SRV/D-T) have been determined. We designed specific primers in the gag region of SRV/D-T that enabled us to directly detect by polymerase chain reaction (PCR) SRV/D-T proviral DNA sequences in DNA extracted from whole blood. Using this assay and another PCR assay that detects multiple SRV/D subtypes, we performed a survey for SRV/D infection in our specific pathogen-free (SPF) and conventional colonies at Tsukuba Primate Center (TPC). In the SPF colony, no SRV/D signal was detected in any animal. On the other hand, SRV/D-T was detected in 11 of 49 animals (22.5%) in the conventional colony. SRV/D-T was the only SRV/D subtype detected. Consequently, SRV/D-T is the major SRV/D subtype present in cynomolgus monkeys at TPC. PMID- 15884777 TI - Ovine model to evaluate ovarian vascularization by using contrast-enhanced sonography. AB - This project was designed to evaluate an ovine model for the use of contrast agent to visualize the microcirculation of normal ovaries. Intraovarian vascularization was investigated in eight ewes by using contrast-enhanced power Doppler after intravenous injection of Sonovue at five different times during each of ten normal estrous cycles. Sheep under general anesthesia underwent transvaginal B mode and power Doppler ultrasound examination of both ovaries, then received two successive doses of 5 ml of Sonovue, one dose for each ovary. Each ovary was monitored after the contrast injection, and a 3-min video clip was stored for each side. The video clip was used to derive time-intensity curves, which were then used to derive the contrast parameters. A total of 108 doses of contrast agent were used; 93% of the injections were available for contrast enhancement analysis. The optimal dose was determined on the first two sheep. Enhancement was strongest and longest with the 5-ml dose. In one sheep, enhancement of both ovaries remained weak irrespective of the period of the cycle. No adverse side effects of Sonovue were seen in the sheep. Contrast injection improved visualization of ovarian microcirculation by 248% (95% confidence interval [CI], 210 to 285%) for 74.2 sec (95% CI, 68.2 to 80.2 sec). Ovaries on the side of ovulation had a stronger enhancement compared with the ovary with no ovulation (368% versus 175%, P < 0.001), but the enhancement time was the same. We concluded that the sheep is an excellent animal model to illustrate microcirculation enhancement by using Sonovue to demonstrate ovarian vascular changes. PMID- 15884778 TI - Reference cardiopulmonary values in normal dogs. AB - The purpose of this project was to collate canine cardiopulmonary measurements from published and unpublished studies in our laboratory in 97 instrumented, unsedated, normovolemic dogs. Body weight; arterial and mixed-venous pH and blood gases; mean arterial, pulmonary arterial, pulmonary artery occlusion, and central venous blood pressures; cardiac output; heart rate; hemoglobin; and core temperature were measured. Body surface area; bicarbonate concentration; base deficit; cardiac index; stroke volume index, systemic and pulmonary vascular resistance indices; left and right cardiac work indices; alveolar partial pressure of oxygen (pO2) ; alveolar-arterial pO2 gradient (A-apO2); arterial, mixed-venous, and pulmonary capillary oxygen content; oxygen delivery; oxygen consumption; oxygen extraction; venous admixture; arterial and mixed-venous blood CO2 contents; and CO2 production were calculated. In the 97 normal, resting dogs, mean arterial and mixed-venous pH were 7.38 and 7.36, respectively; partial pressure of carbon dioxide (pCO2), 40.2 and 44.1 mm Hg, respectively; base deficit, -2.1 and -1.9 mEq/liter, respectively; pO2, 99.5 and 49.3 mm Hg, respectively; oxygen content, 17.8 and 14.2 ml/dl, respectively; A-a pO2 was 6.3 mm Hg; and venous admixture was 3.6%. The mean arterial blood pressure (ABPm), mean pulmonary arterial blood pressure (PAPm), pulmonary artery occlusion pressure (PAOP) were 103, 14, and 5.5 mm Hg, respectively; heart rate was 87 beats/min; cardiac index (CI) was 4.42 liters/min/m2; systemic and pulmonary vascular resistances were 1931 and 194 dynes.sec.cm-5, respectively; oxygen delivery, consumption and extraction were 790 and 164 ml/min/m2 and 20.5%, respectively. This study represents a collation of cardiopulmonary values obtained from a large number of dogs (97) from a single laboratory using the same measurement techniques. PMID- 15884779 TI - Cardiomyopathy in captive owl monkeys (Aotus nancymae). AB - Dilated cardiomyopathy (DCM) was identified histopathologically in a colony of owl monkeys (Aotus nancymae) over a 15-year period. We characterized the incidence of cardiac disease echocardiographically in the colony over a 14-month period. Of 77 monkeys, 21 had systolic myocardial failure, and postmortem examination confirmed the antemortem diagnosis of DCM in eight animals. Monkeys with a questionable diagnosis at the first examination demonstrated progression of disease with time. Left ventricular end-systolic cross-sectional area and left ventricular fractional area change were the indices that most reliably discriminated between normal and diseased animals. Serum cardiac troponin I concentrations were below detectable limits in normal and diseased monkeys. The apparent high prevalence of disease in this colony precluded establishment of reference intervals for Aotus nancymae. Our study provides the first comprehensive echocardiographic evaluation of owl monkeys with cardiomyopathy. PMID- 15884780 TI - Age-related diffuse chronic telogen effluvium- type alopecia in female squirrel monkeys (Saimiri boliviensis boliviensis). AB - We investigated the diffuse alopecia affecting some female squirrel monkeys (Saimiri boliviensis boliviensis) housed in a breeding facility. We randomly selected 100 female and 10 male animals and performed a complete physical exam and a hair assessment on all animals; blood tests, trichograms, hair density; and skin biopsies in representative cases; and a dominance behavioral assessment of 50 animals. Hair coat was normal in 35 female monkeys and all 10 male animals. Of the 65 females with diffuse alopecia, 17 had mild, 22 moderate, and 26 severe hair loss. The alopecia group had a mean age of 9.6 +/- 0.6 years, whereas that of the normal group was 4.7 +/- 0.6 years (P < 0.05). The parity in the alopecia group was 4.2 +/- 0.6 but 2.0 +/- 0.6 (P < 0.05) in the normal group. There were no statistically significant differences in body weight, hemoglobin, blood urea nitrogen, serum glucose, liver aspartate aminotransaminase, or free thyroxine. The trichogram demonstrated 20.8% +/- 1.6% (mean +/- standard error) of telogen hairs in the alopecia group compared with 9.5% +/- 2.8% of the control group (P < 0.05). The hair density in the alopecia group was 52.8 +/- 4.1/cm2 and 79.6 +/- 14.3/cm2 in the control group. Skin biopsies in affected monkeys demonstrated increased telogen follicles, with no fibrosis or inflammation. There were no statistically significant differences in the dominance behavioral analysis. The findings are consistent with chronic telogen effluvium (CTE). A number of organic, behavioral, and dominance-related stress causes of CTE were excluded. CTE appears to be predominantly age-related in this population. CTE in female squirrel monkeys may serve as an animal model for human diffuse alopecia. PMID- 15884781 TI - Susceptibility of rats to corneal lesions after injectable anesthesia. AB - Corneal injury is not a commonly reported side effect after injectable or inhalation anesthesia in rats, but a number of surgery studies at our facility resulted in a high incidence of these injuries. To explore the potential association of various anesthetic protocols with the development of corneal lesions in rats, we retrospectively evaluated clinical records and sections of eyes from 215 male and 187 female Wistar rats used in eight intravenous infusion toxicology studies. None of the studied compounds was associated with eye toxicity. For placement of jugular vein vascular access ports, rats were anesthetized with enflurane, isoflurane, ketamine-xylazine, or Hypnorm-midazolam. Histologically, corneal changes were scored from 0 to 4 in light of degree of mineralization, leukocytic infiltrates, neovascularization, fibrosis, and ulceration. Prestudy (postsurgical) ophthalmic examination findings of corneal opacities were correlated with mineralization of the anterior limiting membrane and corneal ulceration. Corneal lesions were more severe in animals anesthetized with ketamine-xylazine, and minimal changes occurred after anesthesia with either enflurane or isoflurane. The results of further analysis suggest that corneal lesions can be observed within 24 h after injectable anesthetic administration and are not reversible. The severity of corneal changes was reduced when ketamine xylazine anesthesia was reversed with yohimbine. Compared with Sprague-Dawley and Lewis rats, Wistar, Long-Evans, and Fischer 344 rats had increased incidence and severity of corneal lesions after anesthesia with ketamine-xylazine, suggesting that these three strains are at increased risk for developing postanesthetic corneal lesions with this regimen. PMID- 15884782 TI - A national survey of laboratory animal workers concerning occupational risks for zoonotic diseases. AB - In this cross-sectional survey of laboratory animal workers in the United States, 23 of 1367 persons reported 28 cases of infection with zoonotic agents from research animals at their workplace during the past 5 years, with six persons indicating that their infections were medically confirmed. Based on these data, the annualized incidence rate for work-related transmission of zoonotic agents from laboratory animals was 45 cases per 10,000 worker-years at risk (95% confidence interval, 30 to 65 cases), approximating the rate for nonfatal occupational illnesses in the agricultural production-livestock industry and for those employed in the health services during 2002. Logistic regression analysis found various characteristics of persons and their employers that were significantly associated with the likelihood of having been medically evaluated for exposure to a zoonotic agent from laboratory animals. Most (95.595% +/- 1.1%) persons working with laboratory animals or their tissues indicated that they knew whom to talk to at their institution for medical evaluation and care should they be concerned about the possibility of an occupationally acquired zoonotic disease in future. However, occupational illnesses and exposures among laboratory animal workers was underreported, as 10 of the 28 (36%) alleged zoonotic disease cases were not communicated to the employee's supervisor. Lack of concern about the potential significance to their health and the perception of punitive consequences to the employee were some of the reasons cited for underreporting, an issue which must be vigorously addressed in the interests of continuing progress toward zoonotic disease prevention in this field. PMID- 15884783 TI - Successful cyclosporine treatment for atopic dermatitis in a rhesus macaque (Macaca mulatta). AB - A juvenile (1 year old ) female rhesus macaque (Macaca mulatta) developed a chronic active skin condition characterized by pruritus, erythema, alopecia, scaling, exfoliation, and lichenification. Lesions were limited to the ventrum, specifically rostral mandible and neck, axilla and inguinal regions, distal extremities, and interdigital regions. Differential diagnoses included infection, dietary deficiency, metabolic abnormality, endocrinopathy, and immunological injury. Diagnostic tests included complete hemogram, serum chemistry, skin scrapes for ectoparasite detection, hair plucks for dermatophyte culture, and a serum-based hypersensitivity panel. All results were within normal limits. Dermal biopsies revealed lesions consistent with active allergic dermatitis, and a diagnosis of atopic dermatitis was made. Oral cyclosporine (5 mg/kg daily) rapidly eliminated clinical evidence of dermatitis. Histologically, lesions resolved after 12 months of treatment. Atopic dermatitis is an inflammatory skin condition for which there are neither pathognomonic clinical or diagnostic features nor a single successful therapy. Basic criteria such as pruritus, lichenification, a chronic course, and history of allergies strongly support the diagnosis. One successful therapeutic agent is a macrolide calcineurin inhibitor, cyclosporine. It represents a safer class of immunomodulatory drugs than corticosteroids and provides targeted alteration of lymphocyte function. To our knowledge this case represents the first reported successful treatment of atopic dermatitis in a nonhuman primate utilizing cyclosporine. PMID- 15884784 TI - Chromium- and manganese-salen promoted epoxidation of alkenes. PMID- 15884785 TI - Advances in homogeneous and heterogeneous catalytic asymmetric epoxidation. PMID- 15884786 TI - Peptide and amide bond-containing dendrimers. PMID- 15884787 TI - Physical aspects of radiation-induced processes on SiO2, gamma-Al2O3, zeolites, and clays. PMID- 15884788 TI - gem-disubstituent effect: theoretical basis and synthetic applications. PMID- 15884789 TI - Structure-activity relationship studies of chemical mutagens and carcinogens: mechanistic investigations and prediction approaches. PMID- 15884790 TI - Modified BINAP: the how and the why. PMID- 15884791 TI - Conjugated oligoenynes based on the diethynylethene unit. PMID- 15884792 TI - Recent developments in mass spectrometry for the characterization of nucleosides, nucleotides, oligonucleotides, and nucleic acids. PMID- 15884793 TI - Hypoxia inducible factor pathways as targets for functional foods. AB - The etiology of most chronic angiogenic diseases such as rheumatoid arthritis, atherosclerosis, diabetes complications, and cancer includes the presence of pockets of hypoxic cells growing behind aerobic cells and away from blood vessels. Hypoxic cells are the result of uncontrolled growth and insufficient vascularization and have undergone a shift from aerobic to anaerobic metabolism. Cells respond to hypoxia by stimulating the expression of hypoxia inducible factor (HIF), which is critical for survival under hypoxic conditions and in embryogenesis. HIF is a heterodimer consisting of the O2-regulated subunit, HIF 1alpha, and the constitutively expressed aryl hydrocarbon receptor nuclear translocator, HIF-1beta. Under hypoxic conditions, HIF-1alpha is stable, accumulates, and migrates to the nucleus where it binds to HIF-1beta to form the complex (HIF-1alpha + HIF-1beta). Transcription is initiated by the binding of the complex (HIF-1alpha + HIF-1beta) to hypoxia responsive elements (HREs). The complex [(HIF-1alpha + HIF-1beta) + HREs] stimulates the expression of genes involved in angiogenesis, anaerobic metabolism, vascular permeability, and inflammation. Experimental and clinical evidence show that these hypoxic cells are the most aggressive and difficult angiogenic disease cells to treat and are a major reason for antiangiogenic and conventional treatment failure. Hypoxia occurs in early stages of disease development (before metastasis), activates angiogenesis, and stimulates vascular remodeling. HIF-1alpha has also been identified under aerobic conditions in certain types of cancer. This review summarizes the role of hypoxia in some chronic degenerative angiogenic diseases and discusses potential functional foods to target the HIF-1alpha pathways under hypoxic and normoxic conditions. It is reported that dietary quinones, semiquinones, phenolics, vitamins, amino acids, isoprenoids, and vasoactive compounds can down-regulate the HIF-1 pathways and therefore the expression of several proangiogenic factors. Considering the lack of efficiency or the side effects of synthetic antiangiogenic drugs at clinical trials, down-regulation of hypoxia-induced angiogenesis by use of naturally occurring functional foods may provide an effective means of prevention. PMID- 15884794 TI - Detection of irradiated ingredients included in low quantity in non-irradiated food matrix. 1. Extraction and ESR analysis of bones from mechanically recovered poultry meat. AB - Protocol EN 1786 for the detection of irradiated food by electron spin resonance (ESR) spectroscopy was not conceived for the detection of irradiated bone containing ingredients included in low concentration in non-irradiated food. An enzymatic hydrolysis method, realized at 55 degrees C, has been developed for the extraction of the bone fraction. When followed by a purification of the extracts by an aqueous solution of sodium polytungstate, this method made possible the detection of irradiated mechanically recovered poultry meat at very low inclusions (0.5%, wt/wt by ESR) in various meals (quenelles and precooked meals). PMID- 15884795 TI - Detection of irradiated ingredients included in low quantity in non-irradiated food matrix. 2. ESR analysis of mechanically recovered poultry meat and TL analysis of spices. AB - Protocols EN 1786 and EN 1788 for the detection of irradiated food by electron spin resonance spectroscopy (ESR) and thermoluminescence (TL) were not conceived for the detection of irradiated ingredients included in low concentration in nonirradiated food. An enzymatic hydrolysis method, realized at 55 degrees C, has been developed for the extraction of silicate minerals and bone fragments. When followed by a purification of the extracts by an aqueous solution of sodium polytungstate, this method made it possible to detect very low inclusions of irradiated spices (0.05%, wt/wt by TL) included in various meals (cheeses and precooked meals). Even for food containing together two ingredients (spices and mechanically recovered meat), it was possible to detect and identify them simultaneously. PMID- 15884796 TI - Rapid assay for analyzing biogenic amines in cheese: matrix solid-phase dispersion followed by liquid chromatography-electrospray-tandem mass spectrometry. AB - A new rapid and sensitive method based on matrix solid-phase dispersion (MSPD) followed by liquid chromatography-electrospray-tandem mass spectrometry was devised for the determination of biogenic amines at trace levels in cheese samples. The method required 0.25 g of sample, CN-bonded silica as a dispersant sorbent, and a formic acid aqueous solution/methanol mixture as an eluting solvent. Extraction recoveries from soft cheese products were calculated in the 98 +/- 4-110 +/- 6% range. A procedure based on solid-phase extraction was also evaluated for the extraction of these compounds in cheese. Chromatographic separation was performed using a C18 column with an aqueous ammonium acetate/methanol mixture as the mobile phase under gradient conditions. The method was validated in terms of detection limits (LOD), quantitation limits (LOQ), linearity, recovery, precision, and trueness. Results in the 0.05-0.25 mg kg(-1) range were obtained for the LOD of histamine, tyramine, and beta phenylethylamine in soft cheese samples. Linearity was established over 2 orders of magnitude. Excellent precision in terms of intra-day repeatability was calculated (RSD% < 5). The applicability of the method to the determination of biogenic amines in cheese products was demonstrated. PMID- 15884797 TI - Validation of an analytical methodology for determination of oxytetracycline and tetracycline residues in honey by HPLC with fluorescence detection. AB - An analytical method for the determination of OTC and TC residues in honey was developed. Sample treatment involves an extraction in EDTA-McIlvaine buffer, followed by a solid-phase cleanup step. With regard to the cleanup procedure, different SPE cartridges were evaluated and the results presented. The method was validated according to the guidelines laid down by the 2002/657/EC European Decision parameters: decision limit (Cc alpha) and detection capability (CC beta) were 20 and 21 microg/Kg and 49 and 50 microg/Kg for OTC and TC, respectively, and recoveries of OTC and TC from spiked samples, at three fortification levels, were higher than 87% for both compounds. The analytical method was applied to 57 honey samples. PMID- 15884798 TI - Rapid and reliable detection and identification of GM events using multiplex PCR coupled with oligonucleotide microarray. AB - To devise a rapid and reliable method for the detection and identification of genetically modified (GM) events, we developed a multiplex polymerase chain reaction (PCR) coupled with a DNA microarray system simultaneously aiming at many targets in a single reaction. The system included probes for screening gene, species reference gene, specific gene, construct-specific gene, event-specific gene, and internal and negative control genes. 18S rRNA was combined with species reference genes as internal controls to assess the efficiency of all reactions and to eliminate false negatives. Two sets of the multiplex PCR system were used to amplify four and five targets, respectively. Eight different structure genes could be detected and identified simultaneously for Roundup Ready soybean in a single microarray. The microarray specificity was validated by its ability to discriminate two GM maizes Bt176 and Bt11. The advantages of this method are its high specificity and greatly reduced false-positives and -negatives. The multiplex PCR coupled with microarray technology presented here is a rapid and reliable tool for the simultaneous detection of GM organism ingredients. PMID- 15884799 TI - Effects of solvent polarity and acidity on the extraction efficiency of isoflavones from soybeans (Glycine max). AB - Soy isoflavones have been correlated with beneficial health effects. The predominant chemical forms of isoflavones present may affect their biological activities. Choosing the solvent system that can accurately quantify the amounts of individual isoflavones present in these products is paramount. Our objectives were to compare frequently used solvent systems and to evaluate the effects of polarity and acidity on the recovery of isoflavones from soybeans. Isoflavones were extracted from pulverized Manokin soybeans using six solvent systems, which are the combinations of three polarity levels (83% acetonitrile, 80% methanol, and 58% acetonitrile) and two acidity levels (nonacidified and acidified). The pulverized soybean was stirred for 2 h in each solvent system before filtration and concentration using rotary evaporation. The extract was resuspended in 16% acetonitrile and analyzed by high-performance liquid chromatography. Recoveries of pure standards were evaluated with all solvent systems. Solvents with a higher polarity extracted a significantly higher amount of total isoflavones. For individual isoflavones, 58% acetonitrile (highest polarity) extracted either the highest amounts or no less than other solvents, while 83% acetonitrile (lowest polarity) extracted either the lowest amounts or no more than other solvents except for the aglycone form. Acidification significantly reduced the recovery of the malonylglucoside form and the total isoflavones. The recovery study revealed that acidification favored the chemical transformations of isoflavones during the extraction. Among the six solvent systems examined, 58% acetonitrile aqueous solution without acid was the best for extraction of isoflavones from soybeans. PMID- 15884800 TI - Development of a large-scale (50 L) apparatus for ampholyte-free isoelectric focusing (autofocusing) of peptides in enzymatic hydrolysates of food proteins. AB - It has been demonstrated that peptides in enzymatic hydrolysates of proteins can be fractionated on the basis of the amphoteric nature of the sample peptides, by a laboratory-scale isoelectric focusing apparatus, without adding a chemically synthesized carrier ampholyte. This approach is referred to as autofocusing. In the present study, a large-scale (up to 50 L) autofocusing apparatus was developed and tested. A tank (125 cm x 25 cm x 20 cm) was divided into 12 compartments by 11 plates, each with a window covered in a thin agarose gel layer supported by a nylon screen (100 mesh). The compartments at both ends were filled with 0.1 N phosphoric acid (anode) and 0.1 N NaOH (cathode), respectively, functioning as electrode compartments. The remaining compartments were used for sample compartments. Autofocusing was carried out at constant voltage according to two different methods. In method 1, all sample compartments were filled with a 1% water solution of casein or milk whey protein hydrolysates. In method 2, two compartments located in the center of the tank were filled with 5% sample solution and the others were filled with deionized water. Compositional and sequence analyses of the autofocusing fractions revealed that peptides in the two hydrolysates can be fractionated within 24 h by the present apparatus. Better fractionation was obtained by method 2, whereas enrichment of some peptides occurred by using method 1. PMID- 15884801 TI - Determination of polybrominated diphenyl ethers and polybrominated dibenzo-p dioxins/dibenzofurans in marine products. AB - Polybrominated diphenyl ethers (PBDEs) have been widely used as flame retardants in plastics and textile coatings, and these compounds have been recognized as ubiquitous environmental contaminants. Furthermore, it is considered a serious problem that polybrominated dibenzo-p-dioxins and dibenzofurans (PBDD/DFs), having toxicities similar to those of chlorinated dioxins, are generated by the manufacture of brominated flame retardants (BFRs) such as PBDEs, and formed by the combustion of substances containing BFRs. Several congeners of PBDD/DFs and PBDEs have been detected in the adipose tissue of the Japanese. Although food is suspected as an exposure source, little information is available regarding the levels of these brominated compounds in food, as compared with information regarding dioxin or polychlorinated biphenyls. It is necessary to investigate the levels of these brominated organic compounds in various foods and to estimate their influence in the case of human exposure. We developed an efficient method of analyzing PBDEs and PBDD/DFs contents in food samples using accelerated solvent extraction and determined the concentrations in several marine products such as raw fish, processed foods, and seaweed purchased in Japan. A recovery test (n = 5) using the method and involving dried fish showed acceptable recoveries of 57.7-78.5% (RSD 5.4-15.9%) for PBDEs and 50.0-56.4% (RSD 1.5-7.9%) for PBDD/DFs. In the analysis of marine product samples, several congeners of PBDEs were detected in raw fish, processed fish, and seaweed; the highest concentration of sigmaPBDEs was detected in yellowtail (1161 pg/g whole basis), followed by mackerel (553.5 pg/g whole basis). The most dominant congener present in these marine samples was 2,2',4,4'-tetraBDE (#47). PMID- 15884802 TI - Chiral stability of synthetic pyrethroid insecticides. AB - Synthetic pyrethroids are chiral compounds consisting of multiple stereoisomers. Evaluation of enantioselectivity in environmental fate and ecotoxicity requires analytical methods that preserve stereoisomer integrity during analysis. In this study, we characterized the stability of stereoisomers from four commonly used pyrethroids, cis-bifenthrin (cis-BF), permethrin (PM), cypermethrin (CP), and cyfluthrin (CF), during gas chromatography (GC) analysis and sample preparation. Stereoisomers of cis-BF and PM were found to be stable, but those of CP and CF were unstable, under heat or in water. Isomer conversion occurred only at the alphaC in CP or CF, causing the analyte stereoisomer to convert to an epimer. At a GC inlet temperature of 260 degrees C, about 9% conversion occurred for CP and CF. In organic solvents and sterile water, stereoisomers of cis-BF and PM were stable, but slow isomer conversion was observed for CP and CF in water at ambient temperature. However, isomer conversion for CP and CF was relatively insignificant (2-3%) when the GC inlet temperature was kept at < or = 180 degrees C or when on-column injection was used. Isomer conversion at the alphaC in water suggests that abiotic processes may also contribute to enantioselectivity observed in the environment for pyrethroids with the asymmetric alphaC. PMID- 15884803 TI - Discriminating between the six isomers of dicaffeoylquinic acid by LC-MS(n). AB - The fragmentation behavior of all six dicaffeoylquinic acids (diCQA) has been investigated using LC-MS(4). It is possible to discriminate between each of the isomers including those for which commercial standards are not available. For diCQA, the ease of removal of the caffeoyl residue during fragmentation is 1 approximately = 5 > 3 > 4. The distinctive fragmentation observed for the little studied 1,4-dicaffeoylquinic acid involves elimination of the C1 caffeoyl residue, repeated dehydrations leading to the aromatization of the quinic acid moiety, and its decarboxylation. It is suggested that this process is initiated by the C1 carboxyl protonating the C5 hydroxyl in the inverted chair conformer, followed by its protonating the C1 caffeoyl residue in the favored chair conformation. The fragmentation of 1-caffeoylquinic acid is indistinguishable from that of the commercially available 5-caffeoylquinic acid, but these two isomers can be distinguished easily by their facile chromatographic resolution on reversed phase packings. The hierarchical key previously developed for characterizing chlorogenic acids has been extended to accommodate 1 caffeoylquinic acid and the 1-acyl dicaffeoylquinic acids. PMID- 15884804 TI - Event-specific real-time detection and quantification of genetically modified Roundup Ready soybean. AB - The event-specific real-time detection and quantification of Roundup Ready soybean (RRS) using an ABI PRISM 7700 sequence detection system with light upon extension (LUX) primer was developed in this study. The event-specific primers were designed, targeting the junction of the RRS 5' integration site and the endogenous gene lectin1. Then, a standard reference plasmid was constructed that carried both of the targeted sequences for quantitative analysis. The detection limit of the LUX real-time PCR system was 0.05 ng of 100% RRS genomic DNA, which was equal to 20.5 copies. The range of quantification was from 0.1 to 100%. The sensitivity and range of quantification successfully met the requirement of the labeling rules in the European Union and Taiwan. PMID- 15884805 TI - Hapten syntheses and antibody generation for the development of a polybrominated flame retardant ELISA. AB - Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants that are increasingly an environmental concern. Several antibodies were developed for the polybrominated diphenyl ether flame retardant BDE-47 (1), often found in the highest concentration in human milk, plasma, and adipose tissue. Four haptens with different bromine and linker substitution patterns were synthesized and utilized to generate five polyclonal antibodies from goats and two polyclonal antibodies from rabbits. Competition was assessed using four different coating antigens for all seven antibodies. The coating antigen showed marked effects on competition. When the same hapten was used for antibody and the coating antigen less competition was observed. The effect of BDE structure on competition was evaluated by using BDE-47 (1), BDE-99 (2), BDE-100 (3), BDE-153 (4), and BDE-183 (5). None of the compounds showed high competition with antibody I-KLH, presumably because steric hindrance prevented formation of an efficient binding site. As predicted from structural considerations, BDE-47 (1) competed well with the remaining antibodies, whereas BDE-100 (3) competed well with only II-KLH. The remaining congeners (BDE-99 (2), BDE-153 (4), and BDE-183 (5)) contain bromines that cannot be positioned in binding sites and thus cross-react poorly. The competition study demonstrated that a bromine substitution on the congener could occupy a position analogous to the linker's position. PMID- 15884806 TI - Pyrazole derivatives as photosynthetic electron transport inhibitors: new leads and structure-activity relationship. AB - Four series of new pyrazoles, namely, 5 4-carboxypyrazolo-3-tert-butylcarboxamide and 6 4-carboxypyrazolo-3-cyclopropylcarboxamide derivatives and 10 pyrazolo[3,4 d][1,3]thiazine-4-one and 9 pyrazolo[3,4-d][1,3]thiazine-4-thione derivatives, were synthesized and screened as potential inhibitors of photosynthetic electron transport. The structures were confirmed by 1H NMR, elemental, and IR analyses. Their biological activity was evaluated in vitro as the ability to interfere with the light-driven reduction of ferricyanide by isolated spinach chloroplasts. Only a few compounds exhibited excellent inhibitory properties in the micromolar range, comparable to those of commercial herbicides sharing the same target, such as diuron, lenacil, and hexazinone. Nevertheless, most of the remaining molecules exerted a remarkable inhibition in the millimolar range. Combined with previous results on 6 pyrazolo[1,5-a][1,3,5]triazine-2,4-dione and 4 pyrazolo[1,5 c][1,3,5]thiadiazine-2-one derivatives, these data allowed a comprehensive analysis of structure-activity relationship. Molecular modeling studies were undertaken to rationalize the structural determinants of activity in terms of shape, size, and molecular fields. Results suggested that the inhibitory potential of these compounds is associated mainly with their electrostatic properties. PMID- 15884807 TI - Translation of in vitro inhibition by marine natural products of the C4 acid cycle enzyme pyruvate P(i) dikinase to in vivo C4 plant tissue death. AB - Marine organism derived extracts, previously identified as containing compounds that inhibited the C4 acid cycle enzyme pyruvate P(i) dikinase (PPDK), were assessed for their ability to exhibit an effect on the C4 plants Digitaria ciliaris and Echinochloa crus-galli. Oxygen electrode studies revealed that over half of these extracts inhibited C4 acid driven photosynthesis in leaf slices. Seventeen extracts had a deleterious effect on C4 plants in vivo within 24 h, whereas 36 caused an observable phytotoxic response in one or both of the C4 plants used for in vivo testing. None of the extracts inhibited PPDK metabolism of pyruvate via a directly competitive mechanism, instead hindering the enzyme by either mixed or uncompetitive means. This screening strategy, using a suite of assays, led to the isolation and identification of the herbicidal marine natural product ilimaquinone. PMID- 15884808 TI - Concise synthesis of the plant growth regulator theobroxide. AB - A plant growth regulator, theobroxide, which produces potato tubers under noninduced long-day conditions, was synthesized in four steps from dihydrotoluene. PMID- 15884809 TI - Antiinflammatory flavonoids from Artocarpus heterophyllus and Artocarpus communis. AB - The antiinflammatory activities of the isolated flavonoids, including cycloartomunin (1), cyclomorusin (2), dihydrocycloartomunin (3), dihydroisocycloartomunin (4), cudraflavone A (5), cyclocommunin (6), and artomunoxanthone (7), and cycloheterohyllin (8), artonins A (9) and B (10), artocarpanone (11), artocarpanone A (12), and heteroflavanones A (13), B (14), and C (15) from Artocarpus communis and A. heterophyllus, were assessed in vitro by determining their inhibitory effects on the chemical mediators released from mast cells, neutrophils, and macrophages. Compound 4 significantly inhibited the release of beta-glucuronidase and histamine from rat peritoneal mast cells stimulated with P-methoxy-N-methylphenethylamine (compound 48/80). Compound 11 significantly inhibited the release of lysozyme from rat neutrophils stimulated with formyl-Met-Leu-Phe (fMLP). Compounds 8, 10, and 11 significantly inhibited superoxide anion formation in fMLP-stimulated rat neutrophils while compounds 2, 3, 5, and 6 evoked the stimulation of superoxide anion generation. Compound 11 exhibited significant inhibitory effect on NO production and iNOS protein expression in RAW 264.7 cells. The potent inhibitory effect of compound 11 on NO production in lipopolysaccharide (LPS)-activated macrophages, probably through the suppression of iNOS protein expression. PMID- 15884810 TI - Synthesis and antifungal activities of alkyl N-(1,2,3-thiadiazole-4-carbonyl) carbamates and S-alkyl N-(1,2,3-thiadiazole-4-carbonyl) carbamothioates. AB - A series of alkyl N-(1,2,3-thiadiazole-4-carbonyl) carbamates and S-alkyl N (1,2,3-thiadiazole-4-carbonyl) carbamothioates with unsubstituted or monobrominated straight chain alkyl groups were synthesized and evaluated as fungistatic agents against Gibberella zeae and Alternaria kikuchiana. These compounds showed variable antifungal activities at concentrations of 5 and 50 microg/mL. The results showed that antifungal activities depended on the length of the alkyl chain with the optimal chain length of 6-11 carbons. Carbamic acid, (1,2,3-thiadiazole-4-ylcarbonyl)-, hexyl ester (4) showed a strong fungistatic activity against A. kikuchiana at both concentrations, with 90.7 and 54% growth inhibition at 50 and 5 microg/mL, respectively. Carbamic acid, (1,2,3-thiadiazole 4-ylcarbonyl)-, heptyl ester (5); Carbamic acid, (1,2,3-thiadiazole-4-ylcarbonyl) , octyl ester (6); and Carbamic acid, (1,2,3-thiadiazole-4-ylcarbonyl)-, undecyl ester (9) showed strong fungistatic activity against G. zeae at both concentrations. Their growth inhibitions against G. zeae at the concentration of 5 microg/mL were 78, 63, and 59%, respectively. PMID- 15884811 TI - Production of stilbenoids from the callus of Arachis hypogaea: a novel source of the anticancer compound piceatannol. AB - A new source to produce a significant quantity of a naturally occurring polyphenol, piceatannol, was investigated in this study. Both resveratrol and piceatannol are recognized as important ingredients in functional foods due to their beneficial health effects. However, unlike resveratrol, the piceatannol concentration in plants is very low. Thus, calluses of peanuts, an easily obtainable source, were chosen as the material to induce piceatannol production under controlled conditions. To induce resveratrol and piceatannol, calluses were exposed to the ultraviolet (UV) irradiation. Significant quantities of resveratrol and piceatannol were produced by calluses upon UV irradiation in both static and suspension culture conditions. The amounts of piceatannol and resveratrol produced in 1 g of calluses ranged from 2.17 to 5.31 microg and from 0.25 to 11.97 microg, respectively, in static culture. In suspension culture, the amounts of induced piceatannol and resveratrol were somewhat lower. The quantities of induced piceatannol and resveratrol reached a maximum at 18 h after UV irradiation treatment in static culture. In contrast, the levels of resveratrol and piceatannol remained almost constant throughout the experiments in suspension culture. The piceatannol produced by calluses in all studies was much higher than the values reported in the literature, whereas the resveratrol produced was comparable to reported values. PMID- 15884812 TI - Analyses of arbutin and chlorogenic acid, the major phenolic constituents in Oriental pear. AB - The HPLC retention time, photodiode array UV spectrum analysis, and LC/MS results indicated that arbutin and chlorogenic acid are the main phenolic constituents in Oriental pear. The two compounds exist in different organs of the Yali pear, which is one of the major cultivars of Pyrus bretschnrideri. The contents of arbutin in the leaf bud, floral bud, flower, and young fruit were 11.9, 12.4, 8.29, and 9.92 mg/g fresh weight (FW), respectively. Chlorogenic acid amounts in the same organs were 2.26, 3.22, 5.32, and 3.72 mg/g FW, respectively. During development, the concentration of the two compounds in Yali pears was the greatest in young fruit (9.92 mg/g FW of arbutin and 3.72 mg/g FW of chlorogenic acid), and then declined swiftly with fruit growth to less than 0.400 and 0.226 mg/g FW, respectively, in mature fruit. Large differences existed in the distribution of the two compounds in parts of the mature fruit of 14 Oriental pear cultivars. The greatest concentration of arbutin was found in the peel (1.20 mg/g FW), which was 3-5 times greater than that found in the core and 10-45 times greater than the level in the pulp. The concentration of chlorogenic acid in the core was greater than that in the peel. The compounds in 17 cultivars of Oriental pear, including P. bretschnrideri, Pyrus pyrifolia, Pyrus ussuriensis, and Pyrus sinkiangensis, were compared with those in 5 cultivars of Occidental pear (Pyrus communis). The mean concentration of arbutin in the Oriental pear cultivars was 0.164 mg/g FW, greater than the 0.083 mg/g FW found in the Occidental pear cultivars. The greatest arbutin content was 0.400 mg/g FW, found in the Yali pear. However, the mean concentration of chlorogenic acid in the Oriental pear was 0.163 mg/g FW, less than that found in the Occidental pear (0.309 mg/g FW). PMID- 15884813 TI - Physicochemical properties and bioactivity of fungal chitin and chitosan. AB - Chitinous material was extracted from mycelia of Aspergillus niger and Mucor rouxii grown in yeast peptone dextrose broth for 15 and 21 days, respectively. The extracted material was characterized for purity, degree of acetylation, and crystallinity and tested for antibacterial and eliciting properties. The maximum glucosamine level determined in the mycelium of A. niger was 11.10% dw and in the mycelium of M. rouxii was 20.13% dw. On the basis of the stepwise extraction of freeze-dried mycelia, it appeared that M. rouxii mycelia contained both chitin and chitosan, whereas A. niger contained only chitin. The yields of crude chitin from A. niger and M. rouxii were 24.01 and 13.25%, respectively, and the yield of chitosan from M. rouxii was 12.49%. Significant amounts (7.42-39.81%) of glucan were associated with chitinous compounds from both species and could not be eliminated by the extraction method used. The degrees of acetylation were determined to be 76.53 and 50.07% for chitin from A. niger and M. rouxii, respectively, and 19.5% for M. rouxii chitosan. The crystallinity of fungal chitin and chitosan was estimated to be less intense than in corresponding materials from shrimp shells. The extracted chitin and chitosan in a concentration of 0.1% reduced Salmonella Typhimurium DT104 2576 counts by 0.5-1.5 logs during a 4 day incubation in tryptic soy broth at 25 degrees C. Furthermore, all tested chitinous materials from fungal sources significantly reduced lesions caused by Botrytis cinerea and Penicillium expansum in harvested apples. PMID- 15884814 TI - Effects of Brussels sprout juice on the cell cycle and adhesion of human colorectal carcinoma cells (HT29) in vitro. AB - Consumption of Brassica vegetables is associated with a reduced risk of cancer of the alimentary tract in animal models and human populations. We used raw juice extracted from Brussels sprouts rich in the glucosinolate sinigrin to explore the effect of naturally occurring glucosinolate breakdown products on cell cycle progression and apoptosis in human colorectal carcinoma cells (HT29). Juice was prepared from sprout tissue immediately before use, and the glucosinolate breakdown products were determined by gas chromatography mass spectrometry and liquid chromatography mass spectrometry. The cell cycle was analyzed by flow cytometry on detached and adherent cells, and apoptosis was measured in the detached population by annexin V staining. Twenty-four hours after challenge with juice (10 microL/mL), 7-13% of adherent cells had detached from the substratum but the majority (82%) of these cells had not entered apoptosis, whereas only 33% of detached control cells were not apoptotic (p < 0.05). The main glucosinolate breakdown products were as follows: the sinigrin breakdown product, 1-cyano-2,3 epithiopropane (ca. 38 mM); the gluconapin hydrolysis product, 3-butenyl isothiocyanate (ca. 2.2.mM); the glucobrassicin metabolite, ascorbigen (ca. 8 mM); and low concentrations of other indole glucosinolate-derived hydrolysis products such as neoascorbigen and 3,3'-diindolylmethane. A variety of biologically active glucosinolate breakdown products are released by mechanical disruption of raw Brussels sprout tissue, but contrary to previous assumptions, allyl isothiocyanate is not the main compound responsible for the inhibition of cell proliferation. PMID- 15884815 TI - Anthocyanin metabolism in rats and their distribution to digestive area, kidney, and brain. AB - Anthocyanins are present in human diet due to their wide occurrence in fruits and beverages. They possess antioxidant activities and could be involved in several health effects. The aim of this study was to investigate anthocyanin metabolism and distribution in the digestive area organs (stomach, jejunum and liver) and kidney, as well as a target tissue (brain) in rats fed with a blackberry (Rubus fruticosus L.) anthocyanin-enriched diet for 15 days. Identification and quantification of anthocyanin metabolites was carried out by HPLC-ESI-MS-MS and HPLC-DAD, respectively. The stomach exhibited only native blackberry anthocyanins (cyanidin 3-O-glucoside and cyanidin 3-O-pentose), while in other organs (jejunum, liver, and kidney) native and methylated anthocyanins as well as conjugated anthocyanidins (cyanidin and peonidin monoglucuronides) were identified. Proportions of anthocyanin derivatives differed according to the organ considered, with the liver presenting the highest proportion of methylated forms. Jejunum and plasma also contained aglycone forms. In the brain, total anthocyanin content (blackberry anthocyanins and peonidin 3-O-glucoside) reached 0.25 +/- 0.05 nmol/g of tissue (n = 6). The urinary excretion of total anthocyanins was low (0.19 +/- 0.02% of the ingested amount). Thus, organs of the digestive area indicated a metabolic pathway of anthocyanins with enzymatic conversions (methylation and/or glucurono-conjugation). Moreover, following consumption of an anthocyanin-rich diet, anthocyanins enter the brain. PMID- 15884816 TI - Black raspberry extract and fractions contain angiogenesis inhibitors. AB - Targeted therapies, such as agents that inhibit angiogenesis, offer hope as complementary agents in cancer therapy. Angiogenesis-inhibiting agents have the potential for inhibiting tumor growth and limiting the dissemination of metastasis, thus keeping cancers in a static growth state for prolonged periods. Black raspberry (Rubus occidentalis) extract was discovered to be antiangiogenic (0.1% w/v) in a novel human tissue-based in vitro fibrin clot angiogenesis assay. Assay-guided fractionation of a crude black raspberry extract resulted in a highly potent antiangiogenic fraction that accounted for only 1% of the fresh weight of whole black raspberries. At 0.075% (w/v), the active fraction completely inhibited angiogenic initiation and angiogenic vessel growth. Further subfractionation of this active fraction revealed the coexistence of multiple antiangiogenic compounds, one of which has been identified as gallic acid. However, the individual subfractions did not outperform the active whole fraction. These findings suggest that an active black raspberry fraction may be a promising complementary cancer therapy. It is natural and potent enough for manageable dosing regimens. These extracts contain multiple active ingredients that may be additive or synergistic in their antiangiogenic effects. These observations warrant further investigations in animals and human trials. PMID- 15884817 TI - Phenolic acid, tocopherol and carotenoid compositions, and antioxidant functions of hard red winter wheat bran. AB - An electron spin resonance (ESR) spectrometry study was conducted to examine the free radical scavenging properties of bran extracts of Alliance and Wichita wheat using hydroxyl radical (HO*), 2,2-diphenyl-1-picryhydrazyl radical (DPPH*), and superoxide radical anion (O2*-) and their chelating capacities against Cu2+. Also reported is the radical cation 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS*+) scavenging activity, oxygen radical absorbing capacity (ORAC), and chelating property against Fe2+ of the bran extracts measured by the spectrophotometric methods. Significant radical scavenging and chelating capacities were detected in the bran extracts, along with significant levels of phenolic acids, tocopherols, and carotenoids. Ferulic acid, with a concentration range of 130.60-146.38 microg/g, was the predominant phenolic acid in all of the tested bran samples and accounted for approximately 53-67% of total phenolic acids on a weight basis. Total tocopherol concentration ranged from 1.87 to 2.95 micromol/100 g of bran, whereas total carotenoid level was 0.20-0.33 micromol/100 g of bran. In addition, both wheat variety and growing conditions might significantly alter antioxidant properties and concentrations of beneficial components in wheat bran. PMID- 15884818 TI - Use of a simplified HPLC-UV analysis for soyasaponin B determination: study of saponin and isoflavone variability in soybean cultivars and soy-based health food products. AB - Soyasaponins are phytochemicals of major interest for health. Their identification and quantification remain difficult owing to the large number of structural isomers in soybeans and the lack of stable standards. In this study, a rapid method using high performance liquid chromatography (HPLC) using a UV detector (205 nm) was developed to identify and quantify soyasaponins belonging to group B and compare them with isoflavones in different soy materials. 2,3 Dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)-conjugated soyasaponins were determined using external calibration or a molecular mass ratio after alkaline hydrolysis to cleave their DDMP moieties. The detection limit of soyasaponin I, used as a reference molecule to simplify the analysis, was 0.065 micromol/g. Soyasaponin contents in seven soybean varieties ranged from 13.20 to 42.40 micromol/g in the germ and from 2.76 to 6.43 micromol/g in the cotyledons. The within-day and between-days variation coefficients did not exceed 7.9 and 9.0%, respectively, for the major soyasaponins. Soyasaponin B quantification in different soy-based health supplements was reported along with measurements of their isoflavone content to provide information on the variability of these bioactive compounds among different types of soy food materials. PMID- 15884819 TI - Yeast (Saccharomyces cerevisiae) protein concentrate: preparation, chemical composition, and nutritional and functional properties. AB - The main objective of the present study was to compare the composition and functional and nutritional properties of whole yeast cells (WY) from an ethanol distillery with those of a phosphorylated protein concentrate (PPC) prepared from the same cells. Comparisons were also made of PPC with texturized soy protein (TSP) and soy protein isolate (SPI), both acquired in the local market. Yeast (Saccharomyces cerevisiae) is a rich source of protein, soluble fiber, and some minerals. Saturated fatty acids predominated over monounsaturated and polyunsaturated in both WY and PPC. The functional properties of PPC were similar to those of SPI and TSP. Both soy products and PPC replaced 20 or 40% chuck roll protein without affecting the emulsion properties of the meat products. Amino acid scoring was high for both WY and PPC; digestibility was higher (90%) for PPC and lower (68%) for WY. The protein nutritive value of PPC did not differ from that of casein and was significantly higher than that for WY. PMID- 15884820 TI - Monitoring of monooctanoylphosphatidylcholine synthesis by enzymatic acidolysis between soybean phosphatidylcholine and caprylic acid by thin-layer chromatography with a flame ionization detector. AB - Thin-layer chromatography with a flame ionization detector (TLC-FID) was used for monitoring the production of structured phospholipids (ML type: L, long-chain fatty acids; M, medium-chain fatty acids) by enzyme-catalyzed acidolysis between soybean phosphatidylcholine (PC) and caprylic acid. It was found that the structured PC fractionated into two to three distinct bands on both plate thin layer chromatography (TLC) and Chromarod TLC. These three bands represented PC of the LL type, ML type, and MM type, respectively. The TLC-FID method was applied in the present study to examine the influence of enzyme dosage, reaction temperature, solvent amount, reaction time, and substrate ratio (caprylic acid/PC, mol/mol) on formation of ML-type PC in a batch reactor with Thermomyces lanuginosa lipase as the catalyst. The formation of ML-type PC was dependent on all parameters examined except for the substrate ratio. The ML-type PC content increased with increasing enzyme dosage, reaction temperature, solvent amount, and reaction time. The substrate ratio had no significant effect on the formation of ML-type PC within the tested range (3-15 mol/mol). The formation of MM-type PC was observed in some experiments, indicating that acyl migration is taking place during reaction since the lipase is claimed to be 1,3-specific. The TLC-FID method offers a simple and cheap technique for elucidation of product and byproduct formation during enzyme-catalyzed reactions for production of phospholipids containing mixtures of long- and medium-chain fatty acids. PMID- 15884821 TI - Impact of protein size distribution on gluten thermal reactivity and functional properties. AB - Wheat gluten structure was modified in different ways: Disulfide bonds were reduced by sulfitolysis, or protein chains were enzymatically hydrolyzed at three different degrees of proteolysis. A kinetic study of the thermal reactivity of the modified glutens showed that gluten aggregation kinetic was slowed in consequence to the shift of gluten size distribution toward smaller proteins. In contrary to sulfitolysis, proteolysis also affected the gluten reactivity potential because of the formation of numerous nonreactive species. Moreover, the thermally induced browning reaction was greatly enhanced by proteolysis, which increased the amount of free amine residues, substrates of the Maillard reaction. On the contrary, a whitening effect was observed for reduced gluten with bisulfite. Proteolysis was also found to decrease plasticized gluten viscosity, to increase gluten-based materials water solubility, and to enhance gluten adhesiveness properties but to reduce its mechanical performance. Sulfitolysis was considered as a possible way of extending gluten processability by extrusion or injection molding, whereas proteolysis was found to confer enhanced gluten stickiness that suggests new potential end uses of gluten in the pressure sensitive adhesives domain. PMID- 15884822 TI - Effects of hydrophilic plasticizers on mechanical, thermal, and surface properties of chitosan films. AB - Chitosan films were plasticized with four hydrophilic compounds, namely, glycerol (GLY), ethylene glycol (EG), poly(ethylene glycol) (PEG), and propylene glycol (PG). Our objective was to investigate the effect of plasticizers on mechanical and surface properties of chitosan films. The stability of plasticized films was observed by storage for 3 and 20 weeks in an environmental chamber at 50 +/- 5% RH and 23 +/- 2 degrees C. Plasticization improves the chitosan ductility, and typical stress-strain curves of plasticized films have the features of ductile materials, except the film made with 5% PG that exhibits as a brittle polymer and shows an antiplasticization effect. In most cases, the elongation of plasticized films decreases with the storage time, which might be due to the recrystallization of chitosan and the loss of moisture and plasticizer from the film matrix. Although at the beginning the mechanical properties of films made with PG, at high plasticizer concentration, are comparable to those of films made with EG, GLY, and PEG, their stability is poor and they tend to become brittle materials. The surface properties, analyzed by contact angle measurement, reveal that plasticization increases film hydrophilicity. It is found that GLY and PEG are more suitable as chitosan plasticizers than EG and PG by taking into account their plasticization efficiency and storage stability. Furthermore, a plasticizer concentration of 20% (w/w) with GLY or PEG seemingly is sufficient to obtain flexible chitosan film with a good stability for 5 months of storage. PMID- 15884823 TI - Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food. AB - Two previously developed platforms, a multiplex polymerase chain reaction (PCR) and a peptide nucleic acid (PNA) array, the former allowing for the simultaneous detection of five transgenes and two endogenous controls in food and feed matrices and the latter for the assessment of the identity of amplified PCR products, were combined in order to develop a PNA array device for the screening of genetically modified organisms (GMOs) in food. PNA probes were opportunely designed, synthesized, and deposited on commercial slides. The length of the probes as well as the distance of the probes from the surface were evaluated and found to be critical points. The most suitable probes were found to be 15-mer PNAs linked to the slide surface by means of two 2-(2-aminoethoxy)ethoxyacetic acids as spacers. The device was tested on a model system constituted by flour samples containing a mixture of standards at known concentrations of transgenic material, in particular Roundup Ready soybean and Bt11, Bt176, Mon810, and GA21 maize: The DNA was amplified using the specific multiplex PCR method and tested on the PNA array. The method proposed was found to be able to correctly identify every GMO present in the tested samples. PMID- 15884824 TI - Identification of Chinese medicinal fungus Cordyceps sinensis by PCR-single stranded conformation polymorphism and phylogenetic relationship. AB - Fungi belonging to the Cordyceps species have long been used as food and herbal medicines in Asia and are especially popular as commercially available powdered supplements. Despite this acceptance and use, little is known of the phylogenetic relationships of the genus. Presently, the neighbor-joining method based on the ITS1, 5.8S rRNA, and ITS2 regions was used to construct a phylogenetic tree of 17 Cordyceps isolates. Five major groups were evident. Cordyceps sinensis was less closely related to 15 Cordyceps species but shared a closer relationship with Cordyceps agriota. PCR-single-stranded conformational polymorphism was applied to differentiate seven Cordyceps isolates: five were different from those used to construct the phylogenetic tree, based on differences in the internal spacer 2 (ITS2). The length of ITS2, amplified by primers 5.8SR and ITS4, vary between 334 and 400 bp. This segment could be used for intraspecies classification or detection of mutations and represents potential novel means of identification of this fungal genus in herbal medicines and in quality control applications in the fermentation industry. PMID- 15884825 TI - High-pressure effects on the proteolytic enzymes of sea bass (Dicentrarchus labrax L.) fillets. AB - High-pressure processing is a nonthermal technique ensuring food product safety and enabling a longer shelf life. The purpose of this experiment was to evaluate the effect of high pressure on the main proteolytic enzymes involved in fish muscle degradation during storage. Enzymes were extracted with sarcoplasmic proteins from Dicentrarchus labrax sea bass white muscle. Activity of cathepsins B, D, H, and L was quantified in protein extract, whereas calpain activity was evaluated after isolation from its endogenous inhibitor. High-pressure processing up to 500 MPa enhanced the activity of cathepsin B, H, and L, whereas the activity of cathepsin D increased up to 300 MPa and decreased above 300 MPa. With regard to calpain activity, high-pressure processing led to a decrease of activity, which was zero above 400 MPa. We suggest a leading explanation based on simultaneous deactivation of enzymes and an increase of liberation from lysosomes for cathepsins and on dissociation of subunits for calpains. PMID- 15884826 TI - Influence of different drying and aging conditions on saffron constituents. AB - A dehydration postharvesting treatment is necessary to convert Crocus sativus L. stigmas into saffron spice. Three different dehydration treatments were evaluated: dehydration at room temperature; dehydration with hot air at different temperatures (70, 90, and 110 degrees C); and dehydration following traditional processing in Castille-La Mancha (Spain) with three different heating sources (vineshoot charcoal, gas cooker, and electric coil). The time (between 28 and 55 min) and mean temperature (between 54 and 83 degrees C) conditions for traditional dehydration were established for the first time. The highest coloring strength was obtained when saffron was submitted to higher temperatures and lower times. These findings may be supported by the fact that samples dehydrated at high temperature were more porous than those dehydrated at room temperature, as was observed by scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). The higher the temperature during the process, the higher the proportion of trans-crocetin di-(beta-D-gentibiosyl) ester, although trans crocetin (beta-D-glucosyl)-(beta-D-gentibiosyl) and trans-crocetin di-(beta-D glucosyl) ester decrease while cis-crocins did not change significantly. A thermal aging process reveals that the trans-crocetin di-(beta-D-gentibiosyl) ester increases when saffron is resubmitted to a heating treatment before it is decomposed by the extreme conditions. The picrocrocin extinction during the aging process does not imply a consistent generation of safranal. PMID- 15884827 TI - Bioactive compounds of grapefruit (Citrus paradisi Cv. Rio Red) respond differently to postharvest irradiation, storage, and freeze drying. AB - In the present study, the effect of irradiation, storage, and freeze drying on grapefruit bioactive compounds was investigated. Grapefruits were exposed to one of two irradiation doses: 0 (control) or 300 Gy (137Cs, a proposed treatment against fruit flies) and then stored for up to 6 days. At the last storage time point (6 days after harvest), grapefruit pulp from control and irradiated fruits was freeze-dried. Bioactive compounds were extracted from Rio Red grapefruit pulp and analyzed with reverse phase liquid chromatography while volatile compounds were analyzed using gas chromatography. Freeze-dried pulp from irradiated fruits had a higher (P < or = 0.05) flavonoid content (naringin and narirutin) as compared to the freeze-dried pulp from the control fruits. Freeze-drying treatment reduced (P < or = 0.05) the lycopene content, but the reduction (P < or = 0.05) in beta-carotene content occurred only in the control fruit. Reduction in d-limonene and myrcene was observed in the irradiated fruits at 6 days after harvest and in the freeze-dried samples. These results warrant testing of the effect of postharvest treatments and processing on bioactive compounds in functional systems as they have varied effects on different bioactive compounds of grapefruit. PMID- 15884828 TI - Effect of flour minor components on bubble growth in bread dough during proofing assessed by magnetic resonance imaging. AB - Fermentation of dough made from standard flour for French breadmaking was followed by nuclear magnetic resonance imaging at 9.4 T. The growth of bubbles (size > 117 microm) was observed for dough density between 0.8 and 0.22 g cm(-3). Cellular structure was assessed by digital image analysis, leading to the definition of fineness and rate of bubble growth. Influence of composition was studied through fractionation by extraction of soluble fractions (6% db), by defatting (< 1% db) and by puroindolines (Pin) addition (< or = 0.1%). Addition of the soluble fraction increased the dough specific volume and bubble growth rate but decreased fineness, whereas defatting and Pin addition only increased fineness. The role of molecular components of each fraction could be related to dough elongational properties. A final comparison with baking results confirmed that the crumb cellular structure was largely defined after fermentation. PMID- 15884829 TI - Determination of mechanism of flock sediment formation in tea beverages. AB - The mechanism of sediment formation during the storage of green tea beverage was investigated. Green tea extract was separated by Diaion HP-20 column chromatography, and a sediment-formation test was performed. Results showed that at least one compound of the substance causing flock sediment was contained in each of the HP-20 nonadsorbed and adsorbed fractions. From the following fractionations and structure analyses, the substance in the HP-20 adsorbed fraction was determined to be 1-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl-beta-D glucose (strictinin), which is one of the ellagitannins. Strictinin was hydrolyzed to ellagic acid by heat-sterilization processes such as retort sterilization or the ultra-high temperature processing used during the manufacturing of tea beverages. Ellagic acid combined with proteins in the HP-20 nonadsorbed fraction to form an irreversible sediment of green tea beverage; ellagic acid and proteins were confirmed to be present in that sediment. The HP 20 adsorbed fraction contained little strictinin and formed hardly any sediment, suggesting that control of the strictinin content is significant in avoiding sediment formation during the manufacturing process of tea beverages. PMID- 15884830 TI - Alpha-tocopherol oxidation in fish muscle during chilling and frozen storage. AB - The oxidation of alpha-tocopherol (TH) in chilled and frozen fish muscle was determined using high-performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry. TH oxidation byproducts were identified as alpha-tocopherolquinone (TQ), 5,6-epoxy-alpha-tocopherolquinone (TQE1), and 2-3 epoxy-alpha-tocopherolquinone (TQE2). The concentration of TH decreased significantly during storage while those of TQ, TQE1, and TQE2 increased noteworthy. The relative amounts of TH and its oxidized products were significantly related with the extent of oxidation produced in postmortem fish, and the ratio TQ/TH is suggested as an index of oxidative stress in fish muscle. The effect of phenolic antioxidants supplementation on retarding TH oxidation was also studied. Data suggested that the addition of 100 ppm of caffeic acid, hydroxytyrosol, and propyl gallate could regenerate endogenous TH from its oxidized forms resulting in an antioxidant synergy consistent with the reduction of lipid oxidation observed in fish muscle supplemented with phenolic compounds. PMID- 15884831 TI - Quantitation of short-chain glycerol-bound compounds in thermoxidized and used frying oils. A monitoring study during thermoxidation of olive and sunflower oils. AB - Major short-chain glycerol-bound compounds were investigated in olive oil (OO) and conventional sunflower oil (SO) during thermoxidation at 180 degrees C for 5, 10, and 15 h. These compounds included methyl heptanoate (C7:0), methyl octanoate (C8:0), methyl 8-oxo-octanoate (8-oxo-C8:0), methyl 9-oxononanoate (9-oxo-C9:0), dimethyl octanodiate (C8:0 diester), and dimethyl nonanodiate (C9:0 diester), which were analyzed by GC after derivatization of triacylglycerols to fatty acid methyl esters. An acceptable linear correlation (r = 0.967) was found between the total content of these compounds and the total content of polar compounds, suggesting that quantitation of the major short-chain glycerol-bound compounds provides a good indication of the total alteration level of oils heated at frying temperature. Samples with levels of polar compounds around 25% on oil showed total contents within 2-3 mg/g of oil. To determine the content of these compounds in used frying oils, 10 samples from restaurants and fried-food outlets in Spain were analyzed. Results showed total levels between 2.13 and 7.56 mg/g of oil in samples with contents of polar compounds ranging from 18.8 to 55.5% on oil. Samples with levels of polar compounds of approximately 25% showed total contents of the short-chain compounds similar to those found in the thermoxidized oils, that is, within 2-3 mg/g of oil. PMID- 15884832 TI - Stability and sensory shelf life of orange juice pasteurized by continuous ohmic heating. AB - Electrical heating of food products provides rapid and uniform heating, resulting in less thermal damage to the product. The objective of this research was to examine the effects of ohmic heating on the stability of orange juice with comparison to conventional pasteurization. During storage at 4 degrees C, degradation curves of ascorbic acid followed a linear decrease pattern in both ohmic-heated and conventionally pasteurized orange juices. For five representative flavor compounds (decanal, octana, limonene, pinene, and myrcene), higher concentrations were measured during storage in the ohmic-heated orange juice than in conventionally pasteurized juice. Although residual pectin esterase activity remained negligible in both types of juices, particle size was lower in the ohmic-heated orange juice. The sensory shelf life was determined by using the Weibull-Hazard method. Although both thermal treatments prevented the growth of microorganisms for 105 days, the sensory shelf life of ohmic-treated orange juice was >100 days and was almost 2 times longer than that of conventionally pasteurized juice. PMID- 15884833 TI - Influence of ethanol concentration on the extraction of color and phenolic compounds from the skin and seeds of Tempranillo grapes at different stages of ripening. AB - The aim of this paper is to study how grape ripeness and ethanol concentration affect the extraction of color and phenolic compounds from skins and seeds during the maceration/fermentation process. Simulated maceration assays were carried out with the grapes at three stages of berry development (vitis vinifera cv. Tempranillo) and different percentages of ethanol in the maceration media. Both ripeness and ethanol content have a considerable effect on the extraction of color and phenolic compounds. Of these two factors, ripeness increases the extractability most. The presence of ethanol in the medium facilitates anthocyanin and especially proanthocyanidin extraction, but it also decreases copigmentation phenomena, which can decrease the color intensity. The higher the ethanol concentration is in the maceration media, the higher the astringency of proanthocyanidins. PMID- 15884834 TI - High-resolution MAS NMR and chemometrics: characterization of the ripening of Parmigiano Reggiano cheese. AB - We performed a preliminary NMR investigation on grated Parmigiano Reggiano cheese of different ripening ages. Principal component analysis (PCA) and discriminant analysis (DA) were able to successfully group the analyzed samples according to their respective ages. PMID- 15884835 TI - Phenolic composition and antioxidant activities in flesh and achenes of strawberries (Fragaria ananassa). AB - High performance liquid chromatography coupled with diode array and coulometric array detectors were used to characterize and quantify phenolic compounds in achenes and flesh of ripe strawberries (cv Totem and Puget Reliance). Total phenolics and total monomeric anthocyanins were measured and antioxidant activities were evaluated by the FRAP and the ORAC assays. Strawberries contained 1% achenes on a fresh weight basis; however, they contributed to about 11% of total phenolics and 14% of antioxidant activities in strawberries. Ellagic acid, ellagic acid glycosides, and ellagitannins were the main contributors to the antioxidant activities of achenes. The major anthocyanin in flesh was pelargonidin-3-glucoside, whereas achenes consisted of nearly equal amounts of cyanidin-3-glucoside and pelargonidin-3-glucoside. Phenolic content and antioxidant activity of strawberry achenes were reduced by industrial processing. However, the levels were still high and strawberry waste byproduct could thus be a possible source of nutraceuticals or natural antioxidants. PMID- 15884836 TI - Determination of the country of origin of garlic (Allium sativum) using trace metal profiling. AB - A method for determining the country of origin of garlic by comparing the trace metal profile of the sample to an authentic garlic database is presented. Protocols for sample preparation, high-resolution inductively coupled plasma mass spectrometry, and multivariate statistics are provided. The criteria used for making a country of origin prediction are also presented. Indications are that the method presented here may be used to determine the geographic origin of other agricultural products. PMID- 15884837 TI - Specificity of codling moth (Lepidoptera: Tortricidae) for the host plant kairomone, ethyl (2E,4Z)-2,4-decadienoate: field bioassays with pome fruit volatiles, analogue, and isomeric compounds. AB - Codling moth, Cydia pomonella (L.), is a severe pest of apples, pears, and walnuts worldwide, and new approaches for precise monitoring and management would be beneficial. Ninety-two pome fruit volatiles were formulated in 23 distinct blends, of which a single 4-component blend of 10-carbon esters showed the only significant attraction of moths in field bioassays conducted in both walnut and apple orchards. A single constituent of this blend, ethyl (2E,4Z)-2,4 decadienoate--the "pear ester", was the major contributing attractant. The pear ester attracted both male and female moths in combined numbers that were comparable to the attractiveness of conspecific sex pheromone. Structure-activity tests were conducted in a series of orchard trials to determine the specificity of attraction of codling moths to the pear ester kairomone. No analogue 10-carbon alcohols, aldehydes, acetates, or other esters elicited significant moth capture responses. Tests with various analogue esters with alcohol chain length moiety substitutions of the (2E,4Z)-2,4-decadienoic acid elicited differential capture responses, with the ethyl exceeding the propyl, methyl, butyl, and hexyl analogues. The (E,Z) geometric isomers of this series of (2E,4Z)-2,4-decadienoic acid esters far exceeded the attractiveness of the (E,E) isomers. The pear ester is a potent attractant of both males and females, and codling moths are highly discriminating and specific in their structure-activity-based attraction to this pear-derived kairomone. These specificity attributes should allow this host plant kairomone to contribute to new abilities for female monitoring and the potential of development of novel and highly selective control practices that should decrease the current dependence on the use of broad-spectrum insecticides. PMID- 15884838 TI - Effect of household processing and unit-to-unit variability of pyrifenox, pyridaben, and tralomethrin residues in tomatoes. AB - Residue levels of pyrifenox, pyridaben, and tralomethrin were determined in unprocessed and processed tomatoes, grown in a experimental greenhouse, to evaluate the effect of three different household processes (washing, peeling, and cooking) and the "unit to unit" variability of these pesticides in tomatoes. The study was carried out on 11 greenhouse tomato samples collected during a 5 week period in which two successive treatments with the studied pesticides were applied. Residue levels in unprocessed and processed tomato samples were determined by means of ethyl acetate extraction and gas chromatography-electron capture detection determination. The washing processing factor results were 0.9 +/- 0.3 for pyridaben, 1.1 +/- 0.3 for pyrifenox, and 1.2 +/- 0.5 for tralomethrin, whereas the peeling processing factors were 0.3 +/- 0.2 for pyridaben and 0.0 +/- 0.0 for both pyrifenox and tralomethrin. The average loss of water in the tomato pure samples during the cooking process was approximately 50%; the cooking processing factors were 2.1 +/- 0.8 for pyridaben, 3.0 +/- 1.1 for pyrifenox, and 1.9 +/- 0.8 for tralomethrin. The unit-to-unit variability factors were determined on three different greenhouse samples analyzing 10 different units of unprocessed tomatoes from each sample. In all cases, the unit to-unit variability factor results were within the range of 1.3-2.2. PMID- 15884839 TI - Residues and half-life times of pyrethrins on peaches after field treatments. AB - The behavior of pyrethrins and piperonyl butoxide (PB) on peaches has been studied after field treatment. Three experiments were carried out at 1, 5, and 10x the concentration recommended by the manufacturer. In all experiments, the initial deposition was below the maximum residue level (MRL), and the half-life time calculated in the 10x experiment for total pyrethrins within 2.3 days was in agreement with the preharvest interval (PHI) recommended. In a model system, the photodegradation rates of the pyrethrins in three commercial products were compared with pyrethrum pale (PP), with and without the presence of peach waxes. The pyrethrins in formulations containing PB showed higher half-life times but were not influenced by the presence of waxes, whereas in the case of PP that does not contain any PB, photodegradation was significantly affected by the presence of waxes. PMID- 15884840 TI - Determination of antagonism between cyhalofop-butyl and other rice (Oryza sativa) herbicides in barnyardgrass (Echinochloa crus-galli). AB - Herbicide antagonism is defined as the reduction of control of certain weeds as the result of applying mixtures of two or more herbicides. Cyhalofop-butyl, a graminicide used for postemergence grass weed control in rice, is antagonized by some rice herbicides when applied simultaneously. The result of this type of antagonism usually results in decreased control of grass weeds. Research has shown that herbicide antagonism between graminicides and other herbicides may be caused by different mechanisms as the result of activity of the tank-mix partner. Using HPLC, the objective of this experiment was to analyze the fate of cyhalofop butyl in barnyardgrass tissue when applied alone and in combination with halosulfuron, propanil, or triclopyr. Results indicated that absorption of cyhalofop-butyl and hydrolysis to its phytotoxic metabolite, cyhalofop-acid, was rapid and that halosulfuron and triclopyr had no effect. Because of a likely interaction of propanil with an apoplastic esterase enzyme, increased levels of cyhalofop-butyl and cyhalofop-acid were detected in barnyardgrass tissue, indicating that cyhalofop-butyl metabolism was hindered by propanil. PMID- 15884841 TI - Evaluation of cocoa- and coffee-derived methylxanthines as toxicants for the control of pest coyotes. AB - Methylxanthines were quantified in coffee, tea, and chocolate products. Tarajuilie tea from India, cocoa powder, and cocoa nibs contained the highest levels of methylxanthines. Theobromine, caffeine, and theophylline combined in the ratios observed in tea and chocolate were ingested by coyotes. Although both mixtures induced acute toxicity, the symptoms accompanying the chocolate methylxanthine mimic were preferable. Manipulation of the ratios of methylxanthines in the chocolate mimic led to the identification of a 5:1 theobromine/caffeine mixture as a promising coyote toxicant. This mixture was then administered to coyotes using the coyote lure operative device (CLOD). Mortality occurred in every coyote that ingested any portion of the CLOD contents. These results indicate that mixtures of theobromine and caffeine have the potential to be developed into a selective, effective, and socially acceptable toxicant for the control of pest coyotes. PMID- 15884842 TI - Low molecular weight polylactic acid as a matrix for the delayed release of pesticides. AB - Low molecular weight polylactic acid (LMW PLA) was used as a matrix to formulate biodegradable matrix granules and films with bromacil using a melt process. The compatibility of the PLA with bromacil was evaluated. The release characteristics of the formulations were investigated in vitro. The degradation and erosion of the formulations were monitored by pH and gravimetric analysis during the course of release. Various granules and films had similar biphasic release patterns, a delayed release followed by an explosive release. The release rates were independent of bromacil content in the matrix, but varied with the geometry of matrices. The mechanisms of diffusion and erosion were involved in the release. The delayed release of the formulations was dominantly governed by the degradation and erosion of PLA. LMW PLA underwent bulk erosion. LMW PLA-based matrix formulations could thus be useful for the application of pesticides to sensitive targets such as seed treatment. PMID- 15884843 TI - Changes in volatile emissions from apple trees and associated response of adult female codling moths over the fruit-growing season. AB - Odors in the headspace of apple trees were characterized by in situ volatile collections in the orchard. Sixty-two compounds were quantitatively identified with thermal desorption-gas chromatography-mass spectrometry over the complete fruit-growing season. Overall quantities in the headspace of fruit-bearing twig were highest at petal fall and at the beginning of June and August. Interestingly, the latter two periods coincide with the flight maxima of the codling moth, Cydia pomonella, one of the principal pest insects of apple fruit worldwide. Dual-choice bioassays with mated adult female moths in a Y-tube olfactometer showed that the blend of plant-derived volatiles repelled this key pest of apple at petal fall and attracted it from July to mid-August. Single component analysis indicated that benzaldehyde and butyl acetate might contribute to the observed repellent effect, but the constituents accounting for the attractant effect mid-season remain to be further elucidated. The attractant effect clearly originates from the apple fruit and not from the twig with leaves, as bioassays demonstrated conclusively. PMID- 15884844 TI - Influence of soil properties on the adsorption-desorption of sulcotrione and its hydrolysis metabolites on various soils. AB - The retention process of sulcotrione and its two hydrolysis products, 1,3 cyclohexanedione (CHD) and 2-chloro-4-methylsulfonylbenzoic acid (CMBA), were studied in four different soils (Belgium, Landes, Perpignan, and Martinique) under laboratory conditions. Adsorption isotherms were well fitted by a Freundlich relationship. The values of K(fa) ranged from 0.4 to 27.0, and the most adsorbed compound was CHD regardless the soil type. Sulcotrione and CMBA exhibited similar retention behaviors. According to adsorption and desorption results, sulcotrione presents a moderate affinity with soil components; however, its leaching capacity needs to be carefully assessed. Clay content seemed to be the most important factor influencing the adsorption capacity of each compound, whereas organic matter and pH had little influence. The soils were classified according to their adsorption capacity in the following decreasing order: Martinique, Belgium, Landes, Perpignan. PMID- 15884845 TI - Soil pH on mobility of imazaquin in oxisols with positive balance of charges. AB - The influence of soil pH on the leaching potential of the ionizable herbicide imazaquin was assessed on the profile of two highly weathered soils having a net positive charge in the B horizon, in contrast to a soil having a net negative charge in the whole profile, using packed soil column experiments. Imazaquin leached to a large extent and faster at Kd values lower than 1.0 L kg(-1), a much more lenient limit than usually proposed for pesticides in the literature (Kd < 5.0 L kg(-1)). The amount of imazaquin leached increased with soil pH. As the soil pH increased, the percentage of imazaquin in the anionic forms, the negative surface potential of the soils, as well as imazaquin water solubility also increased, thus reducing sorption because of repulsive electrostatic forces (hydrophilic interactions). For all surface samples (0-0.2 m), imazaquin did not leach at soil pH values lower than pKa (3.8) and more than 80% of the applied amount was leached at pH values higher than 5.5. For subsurface samples from the acric soils, imazaquin only began to leach at soil pH values > zero point of salt effects (ZPSE > 5.7). In conclusion, the use of surface K(oc) values to predict the amount of imazaquin leached within soil profiles having a positive balance of charges may greatly overestimate its actual leaching potential. PMID- 15884846 TI - Determining kinetic and nonequilibrium sorption behavior for chlopyrifos using a hybrid batch/column experiment. AB - Pesticide mobility in soil is strongly coupled to the chemical's sorption characteristics. A modified soil column batch experiment was conducted to measure the transient nature of chlorpyrifos sorption and desorption from Cecil soil. This experimental system minimizes many shortcomings associated with obtaining sorption parameters by fitting soil column data to an advective-dispersive transport equation. Several chlorpyrifos formulations were investigated to determine how formulations affect soil sorption, and if this effect is adequately described using transient sorption/desorption algorithms. Both a second-order sorption with first-order desorption kinetic model and the two-site kinetic/equilibrium model were found to yield reasonable comparisons to experimental observations. In general, the formulation temporarily decreases the sorptivity of chlorpyrifos and alters the time for equilibrium to be achieved. Care must be exercised when extrapolating sorption data for a pure molecule in a laboratory setting to formulated materials used in field applications when environmental fate predictions are sought. PMID- 15884847 TI - Effects of incorporated corn residues on glyphosate mineralization and sorption in soil. AB - In modern agricultural systems employing conservation tillage practices, glyphosate is widely used as a preplant burndown herbicide in a wide range of crops. Conservation tillage systems are characterized by a significant presence of crop residues at the soil surface so that glyphosate is applied to a soil matrix rich in poorly decomposed crop residues. Incorporation of corn residues in the range from 0.5 to 4% caused different effects on mineralization and sorption of [14C]glyphosate in sandy and sandy loam soils. More specifically, low levels of incorporated corn residues did not affect or slightly stimulated herbicide mineralization in the sandy and sandy loam soils, respectively. In the sandy soil, incorporation of the highest level of corn residues (4%) caused a decrease in [14C]glyphosate mineralization. [14C]Glyphosate sorption on both soil types was reduced in samples receiving high amounts of incorporated corn residues. PMID- 15884848 TI - Changes in the aroma compounds of sake during aging. AB - Changes in the aroma of sake during aging were investigated by aroma extract dilution analysis (AEDA) and quantitative analysis using the stir bar sorptive extraction method. In AEDA, more odor zones were detected in aged sake than in fresh sake. The dilution factors of aldehydes, polysulfides, and some esters were greater in the aged sake, and their increase during aging was confirmed through a quantitative analysis of sake stored for 0-35 years. Among these compounds, 3 methylbutanal, methional, and dimethyltrisulfide (DMTS) were present in aged sake at concentrations exceeding their odor thresholds, and the highest odor active value was observed for DMTS. Sensory tests showed that supplementation with DMTS contributed to both the total odor intensity and the sulfury odor of aged sake aroma. PMID- 15884849 TI - Antioxidative activity of volatile extracts isolated from Angelica tenuissimae roots, peppermint leaves, pine needles, and sweet flag leaves. AB - Volatile extracts were isolated from dried medicinal plants [Angelica tenuissimae roots (AT, Angelica tenuissima Nakai), peppermint leaves (PL, Mentha arvensis L.), pine needles (PN, Pinus sylvestris L.), and sweet flag leaves (SF, Acorus gramineus Rhizoma)] using steam distillation under reduced pressure, followed by continuous liquid-liquid extraction (DRP-LLE). The extracts were then analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The major volatile constituents of AT, PL, PN, and SF were 3-butylidene-4,5 dihydrophthalide (32 mg/g), menthol (18 mg/g), thunbergol (2.1 mg/g), and cis asarone (37 mg/g), respectively. The inhibitory activity (%) of the extracts against hexanal oxidation ranged from 33 to 98% at a level of 50 microg/mL. Among the volatile extracts, PL and PN increased cell viabilities by 10 and 24%, respectively, at a dose of 1 microg/mL, compared to that of H2O2-treated brain neuroblastoma cells, SK-N-SH. However, a 20% reduction in the malonaldehyde level, an index for lipid peroxidation, was observed at only 1 microg/mL concentration of PN. PMID- 15884850 TI - Reactivity of epicatechin in aqueous glycine and glucose maillard reaction models: quenching of C2, C3, and C4 sugar fragments. AB - Mechanisms of how epicatechin alters the pathways of the Maillard reaction were investigated. Carbon-13 and nitrogen-15 labeling studies were utilized to define the reactivity of epicatechin with glucose, glycine, and/or reaction products in an aqueous model (pH 7, 125 degrees C for 30 min) via GC, GC/MS and HPLC/MS analysis. Quantification of the volatile reaction product isotopomers by GC/MS from a 1:1 labeled to unlabeled glucose (carbohydrate module labeling technique) plus glycine model system indicated the formation of 2,3-butanedione and acetol were primarily formed via intact C4 and C3 sugar fragments, whereas pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine, and cyclotene were primarily formed via intact C2/C2, C2/C3, C3/C3, C3/C3, and C3/C3 sugar fragment pairs, respectively. The formation of these seven compounds was also reported by GC analysis to be dramatically inhibited when epicatechin was added to the glucose/glycine model system (observed 9-113-fold reduction). HPLC/MS analysis of both the glucose-labeled and glycine-labeled model systems with and without epicatechin indicated that epicatechin reacted directly with C2, C3, and C4 sugar fragments, while epicatechin did not report any direct reactivity with glycine. In conclusion, the quenching of sugar fragmentation products via epicatechin was correlated with the observed inhibition on volatile compound formation when epicatechin was added to a glucose/glycine aqueous reaction model system. PMID- 15884851 TI - Thermal degradation studies of food melanoidins. AB - Food melanoidins were isolated from bread crust, coffee, and tomato sauce and their composition was investigated by thermal degradation. Among the generated volatiles, important food flavor compounds were detected: in particular furans, carbonyl compounds, 1,3-dioxolanes, pyrroles, pyrazines, pyridines, thiophenes, and phenols. The results indicated that the isolated melanoidin fractions mainly consisted of compounds formed from carbohydrates and their degradation products. Besides proteins, other food constituents were incorporated in the melanoidin structure as well, such as lipid oxidation products in tomato melanoidins and phenolic compounds in coffee melanoidins. A comparison of the thermal generation of volatiles between these food-derived melanoidins and model melanoidins prepared from a single carbonyl compound and an amino acid showed that the degradation pattern of food melanoidins is quite different from that obtained from a glucose-glycine model system. PMID- 15884852 TI - Effect of harvest time and drying method on biomass production, essential oil yield, and quality of peppermint (Mentha x piperita L.). AB - In the period from 2000 to 2002, studies on peppermint (Mentha x piperita) herb and essential oil (EO) production have been conducted at Planteforsk, Apelsvoll Research Centre Div. Kise in Norway. The trials were aimed at finding the optimal harvest date and suitable drying methods to maximize EO yield and to obtain a desirable oil quality. Peppermint plants from the first production year (2000 and 2001) and the second production year (2002) were harvested during flowering at three developmental stages (early, full, and late bloom). Biomass and leaf production were recorded, and the water content of the plant material was detected after the application of different drying methods: instantaneous drying at 30, 50, and 70 degrees C and prewilting (ground drying) for 1 or 5 days followed by final drying at 30 degrees C. Finally, plant samples were transferred to The Plant Biocentre at NTNU, Trondheim, Norway, for hydrodistillation and gas chromatography-mass spectrometry (GC-MS) analyses of the EOs. Peppermint oil yield increased from early to full bloom and late bloom (average of all years and drying methods except for 50 and 70 degrees C: 2.95, 4.13 and 4.20 L/daa, respectively) as an effect of biomass production and leaf growth. The flavor impact compounds, menthol and menthone, reached their optimum at full bloom (43 54 and 12-30%, respectively). Prewilting led to slight decreased EO levels after 1 day (7.7%) and 5 days of ground drying (1.5%) and no EO quality changes, compared to direct drying at 30 degrees C. The plant weight (H2O content) was drastically decreased to the average under 80 and 45% in all years, thus reducing the energy supply and costs for the necessary final drying step. PMID- 15884853 TI - Activity-guided identification of (S)-malic acid 1-O-D-glucopyranoside (morelid) and gamma-aminobutyric acid as contributors to umami taste and mouth-drying oral sensation of morel mushrooms (Morchella deliciosa Fr.). AB - Although morel mushrooms are widely used as tasty ingredients in savory dishes, knowledge of the key compounds evoking their attractive taste is still very fragmentary. In the present study, taste activity-guided fractionation of an aqueous morel extract by means of the recently developed taste dilution analysis (TDA) enabled the localization of several umami-like-tasting fractions as well as a fraction imparting an intense mouth-drying sensation to the oral cavity. Hydrophilic interaction liquid chromatography (HILIC), LC-MS, and amino acid analysis led to the successful identification of gamma-aminobutyric acid as the chemical inducer of the mouth-drying and mouth-coating oral sensations imparted by the morel extract. Besides the well-known umami-like taste contributors L glutamic acid, L-aspartic acid, and succinic acid, an additional HILIC fraction was isolated and evaluated as tasting umami-like. LC-MS and NMR studies revealed that this fraction consisted of a mixture of (S)-malic acid 1-O-alpha-D glucopyranoside and (S)-malic acid 1-O-beta-D-glucopyranoside, the structure of which could be successfully confirmed by independent synthesis. To the best of our knowledge, this morel-derived glycoside, which we named (S)-morelid, has previously not been reported in any food products. Sensory analysis of aqueous solutions of the compounds identified revealed threshold concentrations of 0.02 mmol/L for the mouth-drying effect of gamma-aminobutyric acid and 6.0 mmol/L for the umami-like, slightly sour taste of (S)-morelid. PMID- 15884854 TI - The addition of Propionibacterium freudenreichii to Raclette cheese induces biochemical changes and enhances flavor development. AB - Two mixtures of Propionibacterium freudenreichii commercial strains were tested as adjunct cultures in pasteurized milk Raclette cheese to investigate the ability of propionibacteria (PAB) to enhance flavor development. Cheese flavor was assessed by a trained sensory panel, and levels of free amino acids, free fatty acids, and volatile compounds were determined. The PAB level showed a 1.4 log increase within the ripening period (12 weeks at 11 degrees C). Eye formation, which was not desired, was not observed in PAB cheeses. PAB fermented lactate to acetate and propionate and produced fatty acids by lipolysis, branched chain volatile compounds derived from isoleucine and leucine catabolism and some esters. One of the experimental cheeses received the highest scores for odor and flavor intensity and was characterized by higher frequencies of detection for some minor notes ("propionic"and "whey" odor, "sweet" taste). PAB can therefore be considered as potential adjunct cultures to enhance or modify cheese flavor development. PMID- 15884855 TI - Concurrent phenomena contributing to the formation of the aroma of wine during aging in oak wood: an analytical study. AB - Red wine was stored in different oak barrels or in stainless steel, and samples were taken for two years to determine 79 aroma compounds. Aging in oak affects 41 compounds. The type of wood affects 11 compounds. At least seven different processes seem to take place concurrently in aroma evolution, and five such processes, affecting 37 compounds, are linked to the oak cask. These are extraction from the wood, oxidation of wine alcohols and amino acids, microbiological formation of ethyl phenols, sorption processes, and condensation of acetaldehyde with polyphenols. The wood can release linear gamma- and delta lactones, beta-damascenone, and ionones. Some compounds are released very fast from wood, which suggests they lie in the external part of the wood. Some extraction profiles are too complex to be explained by physical processes. Finally, the levels of 2,5-dimethyl-4-hydroxy-3(2H)-furanone and 2-ethyl-5-methyl 4-hydroxy-3(2H)-furanone increase even in the reference wine, which suggests the presence of a precursor. PMID- 15884856 TI - Mechanism of production of troponin T fragments during postmortem aging of porcine muscle. AB - Troponin T (TnT) is one of the myofibrillar proteins that is easily degraded during postmortem aging of pork. In this study, we determined the N-terminal amino acid sequences of TnT degradation fragments produced during postmortem aging and by m-calpain hydrolysis. The N-terminal amino acid sequences of TnT fragments produced during postmortem aging were EVHEPEEKPRPKLTAP, EKPRPKLTAPKIPEG, and APKIPEGEKVDF. On the other hand, the N-terminal amino acid sequences of TnT fragments produced by the action of m-calpain were APPPPAEV, EVHEPEEK, and APK. These sequences of degradation fragments could be mapped on fast type TnT isoform 2. The peptide bonds of His37-Glu38 and Thr51-Ala52 in fTnT2 were cleaved during postmortem aging as well as by the calpain hydrolysis; therefore, calpain was concluded to have an important role in TnT degradation during postmortem aging. It was also found that the sourness-suppressing peptide APPPPAEVHEVHEEVH (Okumura et al. Biosci. Biotechnol. Biochem. 2004, 68, 1657 1662) derived from TnT degradation could be produced by the action of calpains on Glu21-Ala22 and His37-Glu38 sites. PMID- 15884857 TI - Protective activities of stilbene glycosides from Acer mono leaves against H2O2 induced oxidative damage in primary cultured rat hepatocytes. AB - In our previous study, we isolated two new hepatoprotective stilbene glycosides, 5-O-methyl-(E)-resveratrol 3-O-beta-D-glucopyranoside (MRA) and 5-O-methyl-(E) resveratrol 3-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside (MRAG), from the methanolic extract of Acer mono leaves. Thereby, we have attempted to elucidate the hepatoprotective mechanism of these compounds, focusing on antioxidative effects, using hydrogen peroxide (H2O2)-injured primary cultures of rat hepatocytes. Both MRA and MRAG showed potent hepatoprotective activities in pretreatment but showed little effects in posttreatment. In addition, they increased the glutathione (GSH) level in the normal control cultures and significantly prevented the depletion of GSH in H2O2-injured primary cultured rat hepatocytes. Moreover, these compounds significantly restored the level of GSH depleted by buthionine sulfoximine or diethylmaleate in the presence or absence of H2O2. Furthermore, these compounds preserved the activities of antioxidant enzymes such as superoxide dismutase, glutathione reductase, and glutathione peroxidase reduced by H2O2 insults. Meanwhile, MRA and MRAG showed moderate scavenging effects with IC50 values of 103.6 and 80.5 microM, respectively, as determined by 1,1-diphenyl-2-picryl-hydrazyl free radical scavenging activity. Taken together, these results suggest that MRG and MRAG exert significant hepatoprotective activities against H2O2-induced hepatotoxicity by maintaining the antioxidative defense system rather than scavenging free radicals. PMID- 15884858 TI - Inhibitory effect on activator protein-1, nuclear factor-kappaB, and cell transformation by extracts of strawberries (Fragaria x ananassa Duch.). AB - The inhibitory effects of strawberry (Fragaria x ananassa Duch.) antioxidant enzymes on tetradecanoylphorbol-13-acetate (TPA) or ultraviolet-B (UVB) induced activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) were studied. The inhibitory effects of strawberry extracts on the proliferation and transformation of human and mouse cancer cells were also evaluated. Strawberries had high activities of glutathione peroxidase, superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase, and glutathione reductase. Strawberry extracts inhibited the proliferation of human lung epithelial cancer cell line A549 and decreased TPA-induced neoplastic transformation of JB6 P+ mouse epidermal cells. Pretreatment of JB6 P+ mouse epidermal cells with strawberry extract resulted in the inhibition of both UVB- and TPA-induced AP-1 and NF-kappaB transactivation. Furthermore, strawberry extract also blocked TPA-induced phosphorylation of extracellular signal-regulated kinases (ERKs) and UVB-induced phosphorylation of ERKs and JNK kinase in JB6 P+ mouse epidermal cell culture. These results suggest that the ability of strawberries to block UVB- and TPA-induced AP-1 and NF-kappaB activation may be due to their antioxidant properties and their ability to reduce oxidative stress. The oxidative events that regulate AP-1 and NF-kappaB transactivation can be important molecular targets for cancer prevention. The strawberries may be highly effective as a chemopreventive agent that acts by targeting the down-regulation of AP-1 and NF-kappaB activities, blocking MAPK signaling, and suppressing cancer cell proliferation and transformation. PMID- 15884859 TI - Covalent binding of the flavonoid quercetin to human serum albumin. AB - Quercetin is an abundant flavonoid in the human diet with numerous biological activities, which may contribute to the prevention of human disease but also may be potentially harmful. Quercetin is oxidized in cells to products capable of covalently binding to cellular proteins, a process that may be important for its biological activities. In the present study, using radiolabeled drug and quantifying the products after electrophoretic separation, proteins to which oxidized quercetin is binding irreversibly were identified. The binding of quercetin to human serum albumin (HSA) in human blood and the effect of stimulation of neutrophilic myeloperoxidase on this binding were also measured. The in vitro binding of quercetin to eight proteins in the presence of catalytic amounts of horseradish peroxidase and hydrogen peroxide was highly selective for HSA. For all proteins the binding was dramatically decreased by reduced L glutathione. In the blood samples, the release of neutrophilic myeloperoxidase by phorbol ester caused a 3-fold increase in the binding of quercetin to HSA. This study shows that quercetin in the presence of peroxidase/hydrogen peroxide covalently links to proteins with a particularly high affinity for HSA and that this also may occur in vivo after exposure to quercetin. This provides further insights into the complex behavior of this major dietary flavonoid. PMID- 15884860 TI - S-sulfonate contents in raw and cooked meat products. AB - S-Sulfonates (R-S-SO3-) are compounds formed by the reaction between the sulfites added to foodstuffs and the disulfide bonds of cystine, peptides, and proteins. The content of S-sulfonates has been determined in raw sausages and burgers (n = 62). The range of variation in the contents of the determined S-sulfonates is very wide and varies between 47 and 267 microg of SO2/g. The degree of formation of S-sulfonates with regard to the determined sulfite (total SO2 + S-sulfonates) is similar in all of the samples and does not seem to be conditioned by the meat compound (chicken or beef) or by the process of elaboration or type of product (burgers or sausages). In grilled burgers (n = 20) significant losses are produced in the levels of the additive in any of its forms. The value for the S sulfonates is 31 +/- 9.8%, 29 +/- 6.6% corresponding to the free sulfite and a very similar percentage to the total sulfite (free + reversibly bound) 28 +/- 6.7%. It is possible that during the cooking process cleavages of some bound compounds occur, releasing SO2 and reacting to form new adducts. PMID- 15884861 TI - Determination of stilbenes in hop pellets from different cultivars. AB - About 30% of the polyphenols in wort and beer derive from hop, but little is yet known about their nutritional impact. The recent discovery of trans-resveratrol and piceid isomers in hop opens new doors to understanding beer health benefits. In the present work, resveratrol was quantified by HPLC-APCI-MS/MS in pellets from 9 different cultivars. Concentrations ranging from 4 to 9 mg/kg trans piceid, from 2 to 6 mg/kg cis-piceid, and up to 1 mg/kg trans-resveratrol were detected. As previously shown for total polyphenols and flavonoids, the lower the alpha-acid content, the higher the total stilbene content. PMID- 15884862 TI - Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts. AB - Liver fibrosis is a chronic liver disease that will further develop to cirrhosis if severe damage continues to form. A potential treatment for liver fibrosis is to inhibit activated hepatic stellate cell (HSC) proliferation and, subsequently, to induce HSC apoptosis. It has been reported that antioxidants are able to inhibit the proliferation of HSCs. In this study, the aqueous extract of spirulina was chosen as the source of antioxidant to investigate the inhibitory effect on the proliferation of HSC. The growth inhibitory effects of aqueous spirulina and chlorella extract on human liver cancer cells, HepG2, were also studied and compared in pairs. Results indicated that the total phenol content of spirulina was almost five times greater than that of chlorella (6.86 +/- 0.58 vs 1.44 +/- 0.04 mg tannic acid equivalent/g of algae powder, respectively). The antioxidant activity of spirulina determined by the ABTS*+ method was higher than chlorella (EC50: 72.44 +/- 0.24 micromol of trolox equivalent/g of spirulina extract vs 56.09 +/- 1.99 micromol of trolox equivalent/g of chlorella extract). Results of DPPH* assay also showed a similar trend as the ABTS*+ assay (EC50: 19.39 +/- 0.65 micromol of ascorbic acid equivalent/g of spirulina extract vs 14.04 +/- 1.06 micromol of ascorbic acid equivalent/g of chlorella extract). The aqueous extracts of these two algae both showed antiproliferative effects on HSC and HepG2, but spirulina was a stronger inhibitor than chlorella. Annexin-V staining showed that aqueous extract of spirulina induced apoptosis of HSC after 12 h of treatment. In addition, the aqueous extract of spirulina triggered a cell cycle arrest of HSC at the G2/M phase. PMID- 15884863 TI - Interactions of whey proteins during heat treatment of oil-in-water emulsions formed with whey protein isolate and hydroxylated lecithin. AB - The interactions of proteins during the heat treatment of whey-protein-isolate (WPI)-based oil-in-water emulsions with and without added hydroxylated lecithin were studied by examining the changes in droplet size distribution and the quantity and type of adsorbed and unadsorbed proteins. Heat treatment at 90 degrees C of WPI emulsions resulted in an increase in total adsorbed protein; unadsorbed beta-lactoglobulin (beta-lg) was the main protein interacting with the adsorbed proteins during the first 10 min of heating, but after this time, unadsorbed alpha-lactalbumin (alpha-la) also associated with the adsorbed protein. In emulsions containing hydroxylated lecithin, the increase in total adsorbed protein during heat treatment was much lower and the unadsorbed beta-lg did not appear to interact with the adsorbed proteins during heating. However, the behavior of alpha-la during heat treatment of these emulsions was similar to that observed in the emulsions containing no hydroxylated lecithin. In the presence of NaCl, the particle size of the emulsion droplets and the quantities of adsorbed protein increased markedly during heating. Emulsions containing hydroxylated lecithin were less sensitive to the addition of NaCl. These results suggest that the binding of hydroxylated lecithin to unfolded monomers or intermediate products of beta-lg reduces the extent of heat-induced aggregation of beta-lg and consequently decreases the interactions between unadsorbed beta-lg and adsorbed protein. This was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of heated whey protein and hydroxylated lecithin solutions. PMID- 15884864 TI - Redox chemistry of red wine. Quantification by an oscillating reaction of the overall antioxidant power as a function of the temperature. AB - The redox and acid-base reactivity of red wines was studied from both the analytical and kinetic standpoint. Four homemade wines, made from Italian red grape varieties of two different vintages, were tested to study the effect of temperature (25 and 37 degrees C) on the overall antioxidant power, through the Briggs-Rauscher oscillating reaction. The reaction was monitored by potentiometry (platinum electrode) and by direct chronometric detection. A reference scale based on the response of gallic acid was also employed, so as to achieve a quantitative evaluation: the novel Briggs-Rauscher antioxidant index (BRAI) was developed to express the overall antioxidant power quantitatively versus the chosen standard molecule. Overall antioxidant power was found to be related to total polyphenol content measured using the Folin-Ciocalteu method: the older wines had a lower antioxidant ability. Total acidity was also estimated indirectly by means of coupled pH-metric/photometric titrations and visible spectrophotometric measurements; it revealed an overlap between acid-base and redox chemistry of red wine. PMID- 15884865 TI - Binding of selected phenolic compounds to proteins. AB - In the context of this study, the noncovalent binding of selected phenolic compounds (chlorogenic, ferulic, and gallic acids, quercetin, rutin, and isoquercetin) to different proteins (human serum albumin, bovine serum albumin, soy glycinin, and lysozyme) was studied with direct (Hummel-Dreyer/size exclusion chromatography) and/or indirect methods (fluorescence absorbance properties of the binding components). In the latter case, the measurement of the phenol binding was achieved by exploiting the intrinsic fluorescence emission properties of quercetin as a probe. From the data obtained, the binding constants and the number of binding sites were calculated. The binding parameters were influenced by different factors, where, e.g., increasing temperature and ionic strength as well as decreasing pH cause a diminished binding. The structures of the proteins as determined by circular dichroism indicate changes in the tertiary structure with the secondary structure remaining intact. PMID- 15884866 TI - Influence of environmental stresses on stability of O/W emulsions containing cationic droplets stabilized by SDS-fish gelatin membranes. AB - Oil-in-water (O/W) emulsions containing small oil droplets (d32 approximately 0.22 microm) stabilized by sodium dodecyl sulfate (SDS)-fish gelatin (FG) membranes were produced by an electrostatic deposition technique. A primary emulsion containing anionic SDS-coated droplets (zeta approximately -40 mV) was prepared by homogenizing oil and emulsifier solution using a high-pressure valve homogenizer (20 wt % corn oil, 0.46 wt % SDS, 100 mM acetic acid, pH 3.0). A secondary emulsion containing cationic SDS-FG-coated droplets (zeta approximately +30 mV) was formed by diluting the primary emulsion with an aqueous fish gelatin solution (10 wt % corn oil, 0.23 wt % SDS, 100 mM acetic acid, 2.00 wt % fish gelatin, pH 3.0). The stabilities of primary and secondary emulsions with the same oil concentration to thermal processing, ionic strength, and pH were assessed by measuring particle size distribution, zeta potential, microstructure, destabilized oil, and creaming stability. The droplets in secondary emulsions had good stability to droplet aggregation at holding temperatures from 30 to 90 degrees C for 30 min, [NaCl] < or = 100 mM, and pH values from 3 to 8. This study shows that the ability to generate emulsions containing droplets stabilized by multilayer interfacial membranes comprised of two or more types of emulsifiers, rather than a single interfacial layer comprised of one type of emulsifier, may lead to the development of food products with improved stability to environmental stresses. PMID- 15884867 TI - Stepwise extraction of proteins and carbohydrates from soybean seed. AB - The stepwise hot water extraction of soybeans, which were extractions in a series of procedures of whole soybean seeds, dehulled and sliced ones, and pressed ones carried out by autoclaving, was investigated to study the localization in the seed and their characteristics. The characteristics of each extraction were studied by HPLC, SDS-PAGE, components analysis, microscopic observation, and effect for some enzymes. Carbohydrates were easier to extract than protein. In the extractions, the ratio of uronic acid per total sugar was constantly about 0.3. A comparison of these extracts, soybean milk, extraction from defatted soybean meal, and soybean milk residues was also carried out, and the characteristics and the localization were investigated. Mid-sized proteins in soybean milk were easy to extract. However, hardly any high molecular weight proteins or high molecular weight carbohydrates were extracted. The proteins and carbohydrates were considered to be localized in the middle lamella and in the protein and/or oil bodies of the cell, and the proteins and carbohydrates were gradually extracted through seed and cell breaking. Gelation was observed only in the boiled extracts from whole seeds. Pepsin and trypsin digests of the high molecular weight protein had inhibitory activity against the angiotensin I converting enzyme. PMID- 15884868 TI - Intake of soy protein isolate alters hepatic gene expression in rats. AB - Soy protein isolate (SPI) can elicit various physiological effects such as cholesterol lowering and antiobesity effects. To examine whether hepatic gene expression is altered by SPI intake, rats were fed an SPI or casein diet for 8 weeks. After 8 weeks of feeding, liver weight and plasma triglyceride and cholesterol levels were significantly lower in the SPI group than in the casein group. Hepatic gene expression was investigated using DNA microarrays. The expression profiles and statistical analysis showed clear and significant differences between the SPI and casein groups (p < 0.05); in the SPI group, 63 genes were up-regulated and 57 genes were down-regulated, most involved in various physiological functions such as lipid metabolism, antioxidant activity, transcriptional regulation, and energy metabolism. Especially in lipid metabolism, the down-regulated genes are related to fatty acid synthesis and the up-regulated genes are related to cholesterol synthesis and steroid catabolism. These results suggest that SPI intake could maintain homeostasis primarily by modulating lipid and energy metabolism. PMID- 15884869 TI - Human gut microbial degradation of flavonoids: structure-function relationships. AB - The relationship between chemical structure and gut microbial degradation rates of 14 flavonoids, flavone, apigenin, chrysin, naringenin, kaempferol, genistein, daidzein, daidzin, puerarin, 7,4'-dihydroxyflavone, 6,4'-dihydroxyflavone, 5,4' dihydroxyflavone, 5,3'-dihydroxyflavone, and 4'-hydroxyflavone, was investigated by anaerobically fermenting the flavonoids with human gut microflora (n = 11 subjects). Degradation rates for the 5,7,4'-trihydroxyl flavonoids, apigenin, genistein, naringenin, and kaempferol, were significantly faster than the other structural motifs. Puerarin was resistant to degradation by the gut microflora. Extensive degradation of flavonoids by gut microflora may result in lower overall bioavailability than those flavonoids that are slowly degraded because rapidly degrading flavonoids are less likely to be absorbed intact. PMID- 15884870 TI - Fumonisin B-glucose reaction products are less toxic when fed to swine. AB - The effects of fumonisin B-glucose reaction products in swine diets was examined. Pigs were fed diets containing 528 micromol of total fumonisin B/kg (FB), 528 micromol of total FB-glucose adducts/kg (FB-G, 122 micromol of unreacted FB/kg), or 0 micromol of total FB/kg for 15 days to test the efficacy of the FB-G reaction products in detoxifying FB. Weight gain in FB pigs was lower than in FB G or controls, which was correlated with feed intake reduction in FB pigs. Serum aspartate aminotransferase, gamma-glutamyltransferase, and total bilirubin in FB pigs were higher than in FB-G or control pigs. Serum sphinganine/shingosine ratios in FB pigs were higher than in FB-G or control pigs. Microscopic examination of tissues from FB pigs showed generalized liver necrosis and apoptosis with marked cellular pleomorphism and disorganized hepatic cords. The liver and kidneys in the FB-G group appeared to be normal. Tissues of controls were free of lesions. Results suggest that dietary FB-G products are less toxic to swine and may provide an detoxification approach in instances of widespread FB grain contamination (p < 0.05). PMID- 15884871 TI - Tissue distribution, elimination, and metabolism of dietary sodium [36Cl]chlorate in beef cattle. AB - Two steers (approximately 195 kg) were each dosed with 62.5 or 130.6 mg/kg body weight sodium [36Cl]chlorate for three consecutive days. All excreta were collected during the dosing and 8 h withdrawal periods. The apparent radiochlorine absorption was 62-68% of the total dose with the major excretory route being urine. Parent chlorate was 65-100% of the urinary radiochlorine; chloride was the only other radiochlorine species present. Similarly, residues in edible tissues were composed of chloride and chlorate with chloride being the major radiolabeled species present. Chlorate represented 28-57% of the total radioactive residues in skeletal muscle; in liver, kidney, and adipose tissues, chlorate ion represented a smaller percentage of the total residues. Chlorate residues in the low dose steer were 26 ppm in kidney, 14 ppm in skeletal muscle, 2.0 ppm in adipose tissue, and 0.7 ppm in liver. These data indicate that sodium chlorate may be a viable preharvest food safety tool for use by the cattle industry. PMID- 15884872 TI - Inhibitory effect of yuzu essential oil on the formation of N nitrosodimethylamine in vegetables. AB - The inhibitory effect of yuzu (Citrus junos Tanaka) essential oil on the formation of N-nitrosodimethylamine (NDMA) in the presence of vegetables (31 species) or saliva was investigated by HPLC. Most vegetable extracts enhanced the formation of NDMA. However, the formation ratio of NDMA in vegetable extracts was decreased by yuzu oil in the range of 59 to 22%. In the presence of yuzu oil and saliva, its ratio ranged between 62 and 24%. These results indicated that yuzu oil inhibited the formation of NDMA even in vegetables and saliva. The contents of ascorbic acid, nitrate, and nitrite in the 31 vegetable species were 0.3-65 mg/100 g, 3-581 mg/100 g, and 10-750 microg/100 g, respectively. Ascorbic acid and nitrite had little effect on the inhibition or formation of NDMA at their intact levels. Nitrate accelerated the formation of NDMA, and the addition of saliva further enhanced it. The mechanism of inhibition of NDMA formation by alpha-terpinene was studied. It was assumed from the results of LC-MS that a new compound formed by the reaction of alpha-terpinene with nitrite would be a derivative of alpha-terpinene with dinitroso groups. The molecular weight of this compound was 194. It is suggested that terpene hydrocarbons in citrus essential oils would contribute to the decrease of NDMA formation. PMID- 15884874 TI - Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. AB - Methods available for the measurement of antioxidant capacity are reviewed, presenting the general chemistry underlying the assays, the types of molecules detected, and the most important advantages and shortcomings of each method. This overview provides a basis and rationale for developing standardized antioxidant capacity methods for the food, nutraceutical, and dietary supplement industries. From evaluation of data presented at the First International Congress on Antioxidant Methods in 2004 and in the literature, as well as consideration of potential end uses of antioxidants, it is proposed that procedures and applications for three assays be considered for standardization: the oxygen radical absorbance capacity (ORAC) assay, the Folin-Ciocalteu method, and possibly the Trolox equivalent antioxidant capacity (TEAC) assay. ORAC represent a hydrogen atom transfer (HAT) reaction mechanism, which is most relevant to human biology. The Folin-Ciocalteu method is an electron transfer (ET) based assay and gives reducing capacity, which has normally been expressed as phenolic contents. The TEAC assay represents a second ET-based method. Other assays may need to be considered in the future as more is learned about some of the other radical sources and their importance to human biology. PMID- 15884875 TI - Measuring antioxidant effectiveness in food. AB - Many new in vitro methods have been developed to evaluate antioxidant activity. Unfortunately, these in vitro methods often correlate poorly with the ability of compounds to inhibit oxidative deterioration of foods because the in vitro assays do not account for factors such as the physical location of the antioxidant, its interaction with other food components, and environmental conditions. To accurately evaluate the potential of antioxidants in foods, models must be developed that have the chemical, physical, and environmental conditions expected in food products. This paper outlines model systems of the evaluation of antioxidants in three types of foods: bulk oil, oil-in-water emulsions, and muscle foods. These model systems are not intended to be inclusive of all possible methods to measure lipid oxidation and antioxidant activity. However, use of these models would allow researchers to more easily compare research results from one paper to another. PMID- 15884876 TI - Potential cell culture models for antioxidant research. AB - The antioxidant activity of pure compounds, foods, and dietary supplements has been extensively studied with the development of many new antioxidant and antioxidant activity assays in recent years. However, these assays, such as total phenolics, total flavonoids, and total antioxidant activity in vitro, do not reflect the cellular physiological conditions and do not consider the bioavailability and metabolism issues. In addition, the mechanisms of action of antioxidants go beyond the antioxidant activity scavenging free radicals in disease prevention and health promotion. Animal models and human studies are expensive and not suitable for initial antioxidant screening of foods and dietary supplements. Therefore, there is a need for cell culture models to access the bioactivity of antioxidants. This paper is an overview of cell culture models for antioxidant research, as reported at the First International Congress on Antioxidant Methods, held in Orlando, FL, June 16-18, 2004, and outlines potential cell culture models for initial antioxidant screening and antioxidant research. PMID- 15884877 TI - Silver nano-inukshuks on germanium. AB - The integration of metallic nanostructures with semiconductors is important for a variety of technological applications. Through an efficient galvanic displacement reaction on germanium, complex silver nanostructures form spontaneously in aqueous conditions at room temperature. The structures, termed nano-inukshuks, are based on stacks of hexagonal metallic structures that grow, initially, parallel to the surface normal of the germanium. TEM, SEM, XPS, XRD, and EDS indicate that the structures are crystalline silver and, based on open cell potential studies, that their nucleation takes place in the first 100 s, followed by growth of the silver structures, most likely through Volmer-Weber growth. PMID- 15884878 TI - Manufacturing of nanochannels with controlled dimensions using protease nanolithography. AB - The feasibility of creating nanometer scale depressions in biological substrates using active enzymes delivered with scanning probe microscopes has been previously demonstrated by us and other groups. Here we present a comprehensive study revealing the dependence of channels dimensions on the parameters of the "writing" process and provide a simple way to precisely control their dimensions. Such nanochannels may be used in nanofluidic biochip applications. PMID- 15884879 TI - Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics: applications in oral cancer. AB - Gold nanoparticles with unique optical properties may be useful as biosensors in living whole cells. Using a simple and inexpensive technique, we recorded surface plasmon resonance (SPR) scattering images and SPR absorption spectra from both colloidal gold nanoparticles and from gold nanoparticles conjugated to monoclonal anti-epidermal growth factor receptor (anti-EGFR) antibodies after incubation in cell cultures with a nonmalignant epithelial cell line (HaCaT) and two malignant oral epithelial cell lines (HOC 313 clone 8 and HSC 3). Colloidal gold nanoparticles are found in dispersed and aggregated forms within the cell cytoplasm and provide anatomic labeling information, but their uptake is nonspecific for malignant cells. The anti-EGFR antibody conjugated nanoparticles specifically and homogeneously bind to the surface of the cancer type cells with 600% greater affinity than to the noncancerous cells. This specific and homogeneous binding is found to give a relatively sharper SPR absorption band with a red shifted maximum compared to that observed when added to the noncancerous cells. These results suggest that SPR scattering imaging or SPR absorption spectroscopy generated from antibody conjugated gold nanoparticles can be useful in molecular biosensor techniques for the diagnosis and investigation of oral epithelial living cancer cells in vivo and in vitro. PMID- 15884880 TI - Tip-state control of rates and branching ratios in atomic manipulation. AB - We report the atomic manipulation properties of two distinct, stable, and reproducible states of a scanning tunneling microscope tip applied to chlorobenzene/Si(111)-(7x7). We show that the tip state influences the rates of (current-driven) molecular desorption and C-Cl dissociation as well as the branching ratio between these processes, but does not change the mediating electronic channel or the required number of electrons. These manipulation properties combined with the imaging properties of the two tip-states suggest the major difference between tip-states is their coupling efficiency to the pi-states of the chlorobenzene molecule. PMID- 15884881 TI - Integration of cell membranes and nanotube transistors. AB - We report the integration of a complex biological system and a nanoelectronic device, demonstrating that both components retain their functionality while interacting with each other. As the biological system, we use the cell membrane of Halobacterium salinarum. As the nanoelectronic device, we use a nanotube network transistor, which incorporates many individual nanotubes in such a way that entire patches of cell membrane are contacted by nanotubes. We demonstrate that the biophysical properties of the membrane are preserved, that the nanoelectronic devices still function as transistors, and that the two systems interact. Further, we use the interaction to study the charge distribution in the biological system, finding that the electric dipole of the membrane protein bacteriorhodopsin is located 2/3 of the way from the extracellular to the cytoplasmic side. PMID- 15884882 TI - Carbon nanotube-metal cluster composites: a new road to chemical sensors? AB - Novel carbon nanotube-metal cluster structures are proposed as prototype systems for molecular recognition at the nanoscale. Ab initio calculations show that already the bare nanotube cluster system displays some specificity because the adsorption of ammonia on a carbon nanotube-Al cluster system is easily detected electrically, while diborane adsorption does not provide an electrical signature. Since there are well-established procedures for attaching molecular receptors to metal clusters, these results provide a "proof-of-principle" for the development of novel, high-specificity molecular sensors. PMID- 15884883 TI - Local electronic structure of single-walled carbon nanotubes from electrostatic force microscopy. AB - An atomic force microscope was used to locally perturb and detect the charge density in carbon nanotubes. Changing the tip voltage varied the Fermi level in the nanotube. The local charge density increased abruptly whenever the Fermi level was swept through a van Hove singularity in the density of states, thereby coupling the cantilever's mechanical oscillations to the nanotube's local electronic properties. By using our technique to measure the local band gap of an intratube quantum-well structure, created by a nonuniform uniaxial strain, we have estimated the nanotube chiral angle. Our technique does not require attached electrodes or a specialized substrate, yielding a unique high-resolution spectroscopic tool that facilitates the comparison between local electronic structure of nanomaterials and further transport, optical, or sensing experiments. PMID- 15884884 TI - Controlling the electronic interaction between a molecular wire and its atomic scale contacting pad. AB - We report a quantitative study on the electronic interaction between a molecular wire and its atomic scale metallic contacting pad. A so-called "reactive Lander" molecule is manipulated using a low-temperature scanning tunneling microscope to form a planar one-end electronic contact. The increase of the STM contrast at the junction location is discussed by means of the electronic interaction between the contacting group of the molecular wire and the end atoms of the nanopad. PMID- 15884885 TI - Highly efficient multiple exciton generation in colloidal PbSe and PbS quantum dots. AB - We report ultra-efficient multiple exciton generation (MEG) for single photon absorption in colloidal PbSe and PbS quantum dots (QDs). We employ transient absorption spectroscopy and present measurement data acquired for both intraband as well as interband probe energies. Quantum yields of 300% indicate the creation, on average, of three excitons per absorbed photon for PbSe QDs at photon energies that are four times the QD energy gap. Results indicate that the threshold photon energy for MEG in QDs is twice the lowest exciton absorption energy. We find that the biexciton effect, which shifts the transition energy for absorption of a second photon, influences the early time transient absorption data and may contribute to a modulation observed when probing near the lowest interband transition. We present experimental and theoretical values of the size dependent interband transition energies for PbSe QDs. We present experimental and theoretical values of the size-dependent interband transition energies for PbSe QDs, and we also introduce a new model for MEG based on the coherent superposition of multiple excitonic states. PMID- 15884886 TI - Filling carbon nanotubes with particles. AB - The filling of carbon nanotubes (CNTs) with fluorescent particles was studied experimentally and theoretically. The fluorescent signals emitted by the particles were visible through the walls of the nanotubes, and the particles inside the tubes were observable with an electron microscope. Taking advantage of the template-grown carbon nanotubes' transparency to fluorescent light, we measured the filling rate of the tubes with particles at room conditions. Liquids such as ethylene glycol, water, and ethylene glycol/water mixtures, laden with 50 nm diameter fluorescent particles, were brought into contact with 500 nm diameter CNTs. The liquid and the particles' transport were observed, respectively, with optical and fluorescence microscopy. The CNTs were filled controllably with particles by the complementary action of capillary forces and the evaporation of the liquid. The experimental results were compared and favorably agreed with theoretical predictions. This is the first report on fluorescence studies of particle transport in carbon nanotubes. PMID- 15884887 TI - Carbon nanotubes loaded with magnetic particles. AB - We describe a simple and versatile technique to produce magnetic tubes by filling carbon nanotubes (CNTs) with paramagnetic iron oxide particles ( approximately 10 nm diameter). Commercial ferrofluids were used to fill CNTs with an average outer diameter of 300 nm made via chemical vapor deposition into alumina membranes. Transmission electron microscopy study shows a high density of particles inside the CNT. Experiments using external magnetic fields demonstrate that almost 100% of the nanotubes become magnetic and can be easily manipulated in magnetic field. These one-dimensional magnetic nanostructures can find numerous applications in nanotechnology, memory devices, optical transducers for wearable electronics, and in medicine. PMID- 15884888 TI - Synthesis of platinum multipods: an induced anisotropic growth. AB - This paper reports a highly effective synthesis of platinum multipods from platinum 2,4-pentanedionate in organic solvents. A trace amount of silver acetylacetonate is used to trigger the nucleation and the anisotropic growth of Pt nanocrystals. The morphologies of Pt multipods made include I- and V-shaped bipods, various types of tripods, and planar and three-dimensional (3D) tetrapods. The 3D Pt tetrapods can be well-defined, resembling those observed for II-VI semiconducting materials, such as CdS and CdSe. Control of morphology and the multipod-to-sphere transitions under various conditions have been systematically studied. A mode of formation based on the induced kinetically controlled growth has been discussed. PMID- 15884889 TI - High frequency scanning gate microscopy and local memory effect of carbon nanotube transistors. AB - We use impedance spectroscopy to measure the high-frequency properties of single walled carbon nanotube field effect transistors (swCN-FETs). Furthermore, we extend scanning gate microscopy (SGM) to frequencies up to 15 MHz and use it to image changes in the impedance of swCN-FET circuits induced by the SGM tip gate. In contrast to earlier reports, the results of both experiments are consistent with a simple RC parallel circuit model of the swCN-FET, with a time constant of 0.3 micros. We also use the SGM tip to show the local nature of the memory effect normally observed in swCN-FETs, implying that nanotube-based memory cells can be miniaturized to dimensions of the order of tens of nm. PMID- 15884890 TI - Carbon nanotube interaction with DNA. AB - We investigate a system consisting of B-DNA and an array of (10,0) carbon nanotubes periodically arranged to fit into the major groove of the DNA. We obtain an accurate electronic structure of the combined system, which reveals that it is semiconducting and that the bands on either end of the gap are derived exclusively from one of the two components. We discuss in detail how this system can be used as either an electronic switch involving transport through both components, or as a device for ultrafast DNA sequencing. PMID- 15884891 TI - Programming supramolecular assembly and chirality in two-dimensional dicarboxylate networks on a Cu(100) surface. AB - We report a comparative study on the 2D self-assembly of two related ditopic benzoic acid species, which have similar shape and endgroups but different backbone symmetry. High-resolution scanning tunneling microscopy data reveal how the symmetry information of molecular building blocks is readily expressed in the resulting chiral or nonchiral supramolecular networks. The underlying square Cu(100) surface steers network orientation and accounts for carboxylate formation, resulting in an unusual intermolecular hydrogen bond motif. Our results demonstrate that symmetry and chiral resolution in 2D supramolecular assembly can be controlled via the design of functional molecules and choice of substrate. PMID- 15884892 TI - Polymer electrolyte gating of carbon nanotube network transistors. AB - Network behavior in single-walled carbon nanotubes (SWNTs) is examined by polymer electrolyte gating. High gate efficiencies, low voltage operation, and the absence of hysteresis in polymer electrolyte gating lead to a convenient and effective method of analyzing transport in SWNT networks. Furthermore, the ability to control carrier type with chemical groups of the host polymer allows us to examine both electron and hole conduction. Comparison to back gate measurements is made on channel length scaling. Frequency measurements are also made giving an upper limit of approximately 300 Hz switching speed for poly(ethylene oxide)/LiClO(4) gated SWNT thin film transistors. PMID- 15884893 TI - Collecting electrospun nanofibers with patterned electrodes. AB - Electrospinning is a simple, versatile, and useful technique for fabricating nanofibers from a rich variety of functional materials. The nanofibers are usually collected as nonwoven mats, in which the fibers are randomly oriented. We have recently demonstrated that the nanofibers can be uniaxially aligned by introducing an insulating gap into the conductive collector. To elucidate the mechanism of alignment, we have systematically studied the effect of the area and geometric shape of the insulating gap on the deposition of fibers. By modeling the electrostatic forces acting on the fiber, it was established that the fibers tended to be oriented along a direction such that the net torque of electrostatic forces applied to the two ends of a discrete segment of the fiber were minimized. By varying the design of electrode pattern, it was possible to control both alignment and assembly of the electrospun nanofibers. PMID- 15884894 TI - Lasing in single cadmium sulfide nanowire optical cavities. AB - The mechanism of lasing in single cadmium sulfide (CdS) nanowire cavities was elucidated by temperature-dependent and time-resolved photoluminescence (PL) measurements. Temperature-dependent PL studies reveal rich spectral features and show that an exciton-exciton interaction is critical to lasing up to 75 K, while an exciton-phonon process dominates at higher temperatures. These measurements together with temperature and intensity dependent lifetime and threshold studies show that lasing is due to formation of excitons and, moreover, have implications for the design of efficient, low threshold nanowire lasers. PMID- 15884895 TI - Charge control in a model biphenyl molecular transistor. AB - We study charge control in a gated 4,4'-biphenyl diradical molecular transistor using ab initio density functional theory calculations. I-V curves and intrinsic gate capacitances were derived. We find charge control in this transistor to be strongly affected by polarization of the sigma-states of the molecule, leading to strong electrostatic coupling of the internal potentials to the source and drain electrodes, and relatively weak coupling to the gate. We suggest that this spatially dependent and anisotropic polarization is an essential element in the operation of molecular transistors. PMID- 15884896 TI - Enumeration of DNA molecules bound to a nanomechanical oscillator. AB - Resonant nanoelectromechanical systems (NEMS) are being actively investigated as sensitive mass detectors for applications such as chemical and biological sensing. We demonstrate that highly uniform arrays of nanomechanical resonators can be used to detect the binding of individual DNA molecules through resonant frequency shifts resulting from the added mass of bound analyte. Localized binding sites created with gold nanodots create a calibrated response with sufficient sensitivity and accuracy to count small numbers of bound molecules. The amount of nonspecifically bound material from solution, a fundamental issue in any ultra-sensitive assay, was measured to be less than the mass of one DNA molecule, allowing us to detect a single 1587 bp DNA molecule. PMID- 15884897 TI - Diameter-dependent growth direction of epitaxial silicon nanowires. AB - We found that silicon nanowires grown epitaxially on Si (100) via the vapor liquid-solid growth mechanism change their growth direction from 111 to 110 at a crossover diameter of approximately 20 nm. A model is proposed for the explanation of this phenomenon. We suggest that the interplay of the liquid-solid interfacial energy with the silicon surface energy expressed in terms of an edge tension is responsible for the change of the growth direction. The value of the edge tension is estimated by the product of the interfacial thickness with the surface energy of silicon. For large diameters, the direction with the lowest interfacial energy is dominant, whereas for small diameters the surface energy of the silicon nanowire determines the preferential growth direction. PMID- 15884898 TI - Monodispersed spherical colloids of Se@CdSe: synthesis and use as building blocks in fabricating photonic crystals. AB - Monodispersed spherical core-shell colloids of Se@Ag(2)Se have been exploited as a chemical template to synthesize Se@CdSe core-shell particles using a cation exchange reaction. A small amount of tributylphosphine could facilitate the replacement of Ag(+) by Cd(2+) in methanol at 50 degrees C to complete the conversion within 150 min. The orthorhombic structure of beta-Ag(2)Se changed to a well-defined wurtzite lattice for CdSe. The CdSe shells could be converted back to beta-Ag(2)Se by reacting with AgNO(3) in methanol at room temperature. Because of the uniformity in size and high refractive index associated with the Se@CdSe core-shell colloids, they could serve as a new class of building blocks to fabricate photonic crystals with wide and strong stop bands. PMID- 15884899 TI - Electrostatic control of ions and molecules in nanofluidic transistors. AB - We report a nanofluidic transistor based on a metal-oxide-solution (MOSol) system that is similar to a metal-oxide-semiconductor field-effect transistor (MOSFET). Using a combination of fluorescence and electrical measurements, we demonstrate that gate voltage modulates the concentration of ions and molecules in the channel and controls the ionic conductance. Our results illustrate the efficacy of field-effect control in nanofluidics, which could have broad implications on integrated nanofluidic circuits for manipulation of ions and biomolecules in sub femtoliter volumes. PMID- 15884900 TI - Magnetic colloidosomes derived from nanoparticle interfacial self-assembly. AB - Based on the interfacial self-assembly of magnetite nanoparticles, we demonstrate the formation of colloidosomes with shells predominantly composed of monolayers of liquid-like, close-packed nanoparticles. The gelation of aqueous phase with agarose leads to robust and water-dispersible nanoparticle colloidosomes, allowing encapsulation of various water soluble materials. The cutoff of the nanoparticle colloidosomes obtained is primarily defined by the nanoparticle size. This controllable permeability should be of great importance for the encapsulation application. PMID- 15884901 TI - DNA multilayer films on planar and colloidal supports: sequential assembly of like-charged polyelectrolytes. AB - Multilayer films comprising solely negatively charged polyelectrolytes were sequentially assembled based on DNA hybridization. Films prepared from alternating layers of two-block homopolymeric nucleotides (polyA(20)G(20)/polyT(20)C(20)) grew linearly with increasing layer number, as verified by quartz crystal microgravimetry, UV-vis spectrophotometry and optical microscopy. Urea treatment of the films induced morphological changes, while exposure to low ionic strength solutions resulted in film disassembly. DNA multilayer films were also formed on silica particles, and DNA hollow capsules were obtained following dissolution of the template core. PMID- 15884902 TI - Surface plasmon interference nanolithography. AB - A new nanophotolithography technique based on the interference of surface plasmon waves is proposed and demonstrated by using computer simulations. The wavelengths of the surface plasmon waves at metal and dielectric interfaces can reach the nanometer scale while their frequencies remain in the optical range. As a result, the resolution of this surface plasmon interference nanolithography (SPIN) can go far beyond the free-space diffraction limit of the light. Simulation results show that one-dimensional and two-dimensional periodical structures of 40-100 nm features can be patterned using interfering surface plasmons launched by 1D gratings. Detailed characteristics of SPIN such as field distribution and contrast are also investigated. PMID- 15884903 TI - Analytical ultracentrifugation for characterizing nanocrystals and their bioconjugates. AB - Analytical ultracentrifugation (AU) provides a general way to probe the polydispersity of nanoparticles and the formation of bioconjugates in solution. Unconjugated gold nanocrystals show sedimentation coefficient distributions that are in agreement with size distributions as measured by TEM. AU is sensitive to the size/shape changes elicited by conjugation, in this case to lactose repressor (LacI). AU data reveal saturating protein concentrations for conjugates that correspond to the measured stoichiometry of the complex under these conditions. PMID- 15884904 TI - Semiconductor nanocrystal quantum dots on single crystal semiconductor substrates: high resolution transmission electron microscopy. AB - We report on high-resolution transmission electron microscope structural studies of InAs colloidal semiconductor nanocrystal quantum dots (NCQDs) on ultrathin GaAs (001) semiconductor single-crystal substrates. We employ a benign method for preparing electron transparent specimens that is suitable for the study of such fragile samples. The image contrast comprises contributions from electron scattering from both the NCs and the GaAs substrate. Long-term electron exposure studies reveal different damage mechanisms operative in the nanocrystals and the substrate. PMID- 15884905 TI - Versatile visualization of individual single-walled carbon nanotubes with near infrared fluorescence microscopy. AB - Fluorescence microscopy in the near-infrared between 950 and 1600 nm has been developed as a novel method to image and study single-walled carbon nanotubes (SWNTs) in a variety of environments. Intrinsic photoluminescence of disaggregated pristine SWNTs was excited by a diode laser and detected with a two dimensional InGaAs photodiode array. Individual nanotubes were visualized with a spatial resolution of ca. 1 microm and characterized with polarization measurements and emission spectroscopy. Spatially resolved emission spectra allowed (n,m) identification of single nanotubes and revealed small environmentally induced spectral shifts between segments of long tubes. Nanotube motions in aqueous surfactant were visualized with a time resolution of 50 ms and used to estimate the diffusion coefficient. PMID- 15884906 TI - Size-dependent photoconductivity in MBE-grown GaN-nanowires. AB - We report on electrical transport in the dark and under ultraviolet (UV) illumination through GaN nanowhiskers grown by molecular beam epitaxy (MBE), which is sensitively dependent on the column diameter. This new effect is quantitatively described by a size dependent surface recombination mechanism. The essential ingredient for the interpretation of this effect is a diameter dependent recombination barrier, which arises from the interplay between column diameter and space charge layer extension at the column surface. PMID- 15884907 TI - Plasmon polaritons in finite-length metal-nanoparticle chains: the role of chain length unravelled. AB - While the conceptual framework for nanoplasmonic waveguides composed of a chain of noncontacting metal nanoparticles usually neglects the effects of the ends, the long-range nature of the interparticle coupling underlying the electromagnetic transport means that finite chain length can play an important role. Here, the complex energies of the plasmon-polariton modes in finite-length nanoparticle chains are calculated to ascertain the effects of chain length on the mode dispersion and the radiative contribution to the attenuation. The results indicate that, for typical parameters, the infinite-chain limit is reached with approximately 10 nanoparticles. Thus, even for chain lengths well exceeding the attenuation length, long-range coupling of distant nanoparticles is shown to impact the dispersion and radiative loss. PMID- 15884908 TI - Self-organized inorganic nanoparticle arrays on protein lattices. AB - Cavities formed by proteins have been utilized as the reaction chamber for the fabrication of a range of inorganic nanoparticles, providing control of the size of particles by limiting growth and preventing agglomeration. In crystal form, proteins construct molecular arrays that can provide regularly arranged sites for nanoparticles. Here we report the fabrication of nanometric iron and indium particles using ferritin, an iron-storage protein. The indium nanoparticles thus formed have uniform spherical shape with diameter of 6.6 +/- 0.5 nm, while the iron nanoparticles are somewhat irregular in shape (5.8 +/- 1.0 nm). Regular two dimensional arrays of these nanoparticles are successfully produced by crystallizing ferritin molecules on a water-air interface using the denatured protein film method. The lattice constant of these nanoparticle arrays is 13 nm with hexagonal packing, and arrays of more than 1 microm in area can be obtained by transfer onto silicon wafer. PMID- 15884910 TI - Deoxyribozyme-based ligase logic gates and their initial circuits. AB - A complete set (YES, NOT, AND, and ANDNOT) of molecular scale logic gates based on ligase deoxyribozymes was constructed. The activity of these gates was visualized through the formation of cascades with downstream phosphodieseterase YES gates, which performed fluorogenic cleavage. PMID- 15884911 TI - Carbon nanotubes with covalently linked porphyrin antennae: photoinduced electron transfer. AB - Single- and multiwalled carbon nanotubes have been covalently functionalized with free-base porphyrin. The quantity of porphyrin linked to the surface was determined from thermogravimetric and UV-vis analysis. A reversible protonation equilibrium between the attached porphyrin and the residual acid groups of the carbon nanotubes has been identified. Steady-state fluorescence emission spectrum of the solutions of porphyrin-linked carbon nanotubes shows that the porphyrins act as energy absorbing and electron transferring antennae, and the carbon nanotubes act as efficient electron acceptors. The porphyrin-linked carbon nanotubes show 95-100% emission quenching, indicating a fast photoinduced electron transfer. PMID- 15884912 TI - Long-range ordered thin films of block copolymers prepared by zone-casting and their thermal conversion into ordered nanostructured carbon. AB - Large-scale alignment of lamellae in thin films of diblock copolymers containing polyacrylonitrile and poly(n-butyl acrylate) was achieved by casting copolymer solution on a silicon substrate moved away at a constant speed from the casting nozzle (zone-casting). Grazing incidence small-angle X-ray scattering revealed that the lamellae, which were perpendicular to the substrate, were also aligned over macroscopic scale in the direction perpendicular to the casting direction. Such long-range ordered block copolymer films were then converted by pyrolysis into nanostructured carbons, with excellent preservation of lamellar morphology and orientation. PMID- 15884913 TI - Stereoselective enolizations mediated by magnesium and calcium bisamides: contrasting aggregation behavior in solution and in the solid state. AB - The reactions of magnesium and calcium bis(hexamethyldisilazide) with propiophenone have been studied with a view to determine the utility of these bases in the stereoselective enolization of ketones and to uncover the nature of the metal enolate intermediates produced. Both base systems are highly Z selective when the reactions are conducted in the presence of polar solvents. However, in situ monitoring of the magnesium system in arene solution revealed a preference for E-enolate formation, which was confirmed by silyl enol ether trapping studies. Solution NMR studies of the magnesium system in toluene-d8 show the presence of a monomer-dimer equilibrium for the intermediate amidomagnesium enolates. This assignment is supported by the characterization of a disolvated amidomagnesium enolate dimer by crystallographic analysis. Comparative studies of the calcium system show distinctly different behavior. This is exemplified by the characterization of a novel solvent-separated ion pair complex and a monomeric amidocalcium enolate in the solid state. Solution NMR studies of the calcium system in pyridine-d5 reveal the co-existence of the heteroleptic amidocalcium enolate, the bisamide, the bisenolate and the ion pair complex. PMID- 15884914 TI - Selective functionalization of In2O3 nanowire mat devices for biosensing applications. AB - A strategy to covalently attach biological molecules to the electrochemically active surface of indium oxide nanowire (In2O3 NW) mat devices is presented. A self-assembled monolayer (SAM) of 4-(1,4-dihydroxybenzene)butyl phosphonic acid (HQ-PA) was generated on an indium tin oxide (ITO)-coated glass and In2O3 NWs surface. The chemical steps required for surface derivatization were optimized on an ITO surface prior to modifying the In2O3 NWs. The hydroquinone group contained in the HQ-PA SAM was electrochemically oxidized to quinone (Q-PA) at +330 mV. The monolayer of Q-PA was allowed to react with a thiol-terminated DNA. The DNA was paired to its complementary strand tagged with a fluorescence dye. Attachment of DNA was verified using fluorescence microscopy. A device was subsequently prepared on a SiO2-supported mat of In2O3 NWs by depositing gold electrodes on the mat surface. The reaction strategy optimized on ITO was applied to this In2O3 NW-based device. Arrays of In2O3 NWs on a single substrate were electrochemically activated in a selective manner to Q-PA. Activated In2O3 NWs underwent reaction with HS-DNA and gave a positive fluorescence response after pairing with the dye DNA. The unactivated In2O3 NWs gave no response, thus demonstrating selective functionalization of an In2O3 NW array. This can be considered a key step for the future fabrication of large-scale, inexpensive, nanoscale biosensors. PMID- 15884915 TI - Diastereoselective [4+2] annulation of vinyl carbodiimides with N-alkyl imines. Asymmetric synthetic access to the batzelladine alkaloids. AB - A diastereoselective [4+2] annulation of vinyl carbodiimides with chiral N-alkyl imines has been developed to access the stereochemically rich tricyclic core of the batzelladine alkaloids. Its application to the asymmetric synthesis of batzelladine D permitted the use of long-range, directed hydrogenation and stereoselective intramolecular iodoamination as additional key steps to establish the remaining stereocenters within the natural product with excellent stereocontrol. PMID- 15884916 TI - Mild oxidative one-carbon homologation of aldehyde to amide. AB - One-carbon homologation of aldehyde into amide is realized in one-pot by its reaction with potassium alpha-p-methoxyphenyl-alpha-isocyano acetic acid (1c) and hydrochloride salt of dimethylamine (3a) in toluene at room temperature followed by acidic workup. In this multicomponent reaction, 1c served as donor of the CONH2 function to aldehyde, while the dimethylamine acted as a shuttle molecule to initiate/terminate the sequence and to mediate the internal redox process of one of the three-component adducts. Ready accessibility, nominal cost of the reagents, and mild conditions are attractive features of the present method. PMID- 15884917 TI - Parallel transformations of cyclohexene mediated by the Cp*W(NO) fragment. AB - Hydrogenolysis of Cp*W(NO)(CH2CMe3)2 at room temperature in cyclohexene results in the formation of the intermediate 16e organometallic complex, [Cp*W(NO)(eta2 cyclohexene)]. This intermediate leads to three parallel transformations of cyclohexene, namely (a) C-H activation of cyclohexene to form an eta3 cyclohexenyl hydrido complex, (b) combination of cyclohexene and H2 to form a cyclohexyl hydrido complex, and (c) coupling of two molecules of cyclohexene with concomitant loss of two hydrogen atoms to form a complex containing a novel eta1,eta3-(cyclohexyl)cyclohexenyl ligand. Single-crystal X-ray crystallographic analyses of products resulting from transformations (b) and (c) have been effected. PMID- 15884918 TI - "Cation pool" method based on C-C bond dissociation. Effective generation of monocations and dications. AB - The "cation pools" of alkoxyarylcarbenium ions were effectively generated by the oxidative C-C bond dissociation using low temperature electrolysis. The present method is especially effective for the generation and accumulation of dications, which react with carbon nucleophiles. PMID- 15884919 TI - Nickel-catalyzed intermolecular alkyne insertion into cyclobutanones. AB - Cyclobutanones reacted with alkynes in the presence of nickel(0) catalysts to produce cyclohexenones. Oxidative cyclization of the carbonyl group of the cyclobutanone and the alkyne with the nickel(0) was followed by beta-carbon elimination from the resulting oxanickelacyclopentene and subsequent reductive elimination. This reaction achieves a formal alkyne insertion between the carbonyl carbon and the alpha-carbon, providing a six-membered carbocyclic skeleton. PMID- 15884920 TI - Mesoporous silica composites containing multiple regions with distinct pore size and complex pore organization. AB - Porous ceramics are of great interest for filtration, catalysis, and reactive separation processes. Performance in these applications is highly dependent on features such as pore size distribution and connectivity and wall composition. Here, we describe a method allowing the rational design and synthesis of mesoporous silica composites with controlled heterogeneous pore architectures and demonstrate its validity by producing structures with predetermined placement of regions having different pore size and pore organization. PMID- 15884921 TI - Amplification of changes of a thin film's macromolecular structure into macroscopic reaction-diffusion patterns. AB - A reaction-diffusion process induced from a micronetwork geometry amplifies changes in the molecular structure of a thin gel film into macroscopic readout patterns. When the gel undergoes a helix-to-coil phase transition, the patterns formed by RD switch from symmetry-broken to symmetric ones. Theoretical analysis explains how the system reconfigures internally in response to mass transfer between the applied network and the probed film. PMID- 15884922 TI - Photoisomerization of cis-1-(2-naphthyl)-2-phenylethene in methylcyclohexane at 77 K: no hula-twist. AB - The cis-trans photoisomerization of cis-1-(2-naphthyl)-2-phenylethene (c-NPEA and c-NPEB) was studied in methylcyclohexane glass at 77 K. The progress of the reaction was followed by fluorescence spectroscopy. Formation of the extended trans conformer (t-NPEB) is revealed by the growth of sharp vibronic bands on the broad structureless fluorescence spectrum of c-NPE. Principal component analysis with self-modeling of a matrix consisting of spectra generated by irradiation of c-NPE at different excitation wavelengths and spectra of t-NPE measured under the same conditions as a function of excitation wavelength reveals that the diabatic photoisomerization is a completely conformer-specific one-bond twist process: c NPEB --> t-NPEB. These observations are consistent with observations in solution at ambient temperature. They demonstrate unequivocally that free volume restrictions imposed by the amorphous glassy environment do not open a hula-twist process. PMID- 15884923 TI - Multiple-steering QM-MM calculation of the free energy profile in chorismate mutase. AB - A novel technique for computing free energy profiles in enzymatic reactions using the multiple steering molecular dynamics approach in the context of an efficient QM-MM density functional scheme is presented. The conversion reaction of chorismate to prephenate catalyzed by the Bacillus subtilis enzyme chorismate mutase has been chosen as an illustrative example. PMID- 15884924 TI - Strong attraction among the fully hydrophilic {Mo72Fe30} macroanions. AB - We report the study on the unique driving forces of the self-assembly of fully hydrophilic, soluble {Mo72Fe30} macroanions into single-layer, vesicle-like "blackberry" structures in water and mixed solvents. The hydrophobic interaction that is responsible for the vesicle formation of amphiphilic surfactants does not contribute to the current blackberry formation because of the absence of hydrophobic moiety. The hydrogen bond, van der Waals force, and chemical interaction only play minor roles. Laser light scattering and conductance measurements on a series of {Mo72Fe30}/ethanol/H2O solutions show that a certain amount of negative charges are necessary for the self-assembly, clearly indicating the existence of long-range attraction between macroanions, presumably due to the small counterions in between. The experimental results suggest that the charges on macroanions play a dual effect: short-range electrostatic repulsion and long-range "like-charge attraction", which is the major source of attractive force between hydrophilic macroanions, while van der Waals force, hydrogen bonds, and temporary inter-{Mo72Fe30} Fe-O-Fe chemical linking may also have minor contributions. PMID- 15884925 TI - Distinct mechanisms for the oxidative addition of chloro-, bromo-, and iodoarenes to a bisphosphine palladium(0) complex with hindered ligands. AB - We report an example of a bisphosphine palladium(0) complex with hindered ligands that undergoes oxidative addition of chloro-, bromo-, and iodoarenes in high yield. Addition of PhX (X = I, Br, Cl) to [Pd(Q-phos-tol)2] produced [Pd(Q-phos tol)(Ph)(I)], [Pd(Q-phos-tol)(Ph)(Br)], and [Pd(Q-phos-tol)(Ph)(Cl)]2. To study the mechanisms of the oxidative addition of the three haloarenes to [Pd(Q-phos tol)2], we determined the order of the reaction on the concentration of ligand and haloarene. The different haloarenes reacted through different mechanistic pathways. Addition of iodobenzene occurred by irreversible associative displacement of a phosphine. Addition of bromobenzene occurred by rate-limiting dissociation of phosphine. Addition of chlorobenzene occurred by reversible dissociation of phosphine, followed by rate-limiting oxidative addition. The mechanism of exchange of ligands from the Pd(0)L2 was also studied. The rate constant value for dissociation of ligand calculated from ligand exchange experiments is in agreement with the value calculated through experiments on oxidative addition. PMID- 15884926 TI - One-pot regio- and stereoselective cyclization of 1,2,n-triols. AB - A simple and efficient process to cyclize triols containing a 1,2-diol functionality with a pendant hydroxyl group is presented. The one-pot procedure converts the 1,2-diol into an ortho ester in situ, which upon treatment with a Lewis acid generates a cyclic acetoxonium intermediate. This intermediate is subsequently trapped by the pendant hydroxyl group to generate a cyclic ether. The stereochemistry of the 1,2-diol is transferred to the product with complete fidelity (inversion at the site of cyclization), and the reaction proceeds with high regioselectivity. The process is akin to the Lewis acid-catalyzed intramolecular ring-opening of epoxides with hydroxyl groups yielding cyclic ethers of various sizes with regio- and stereochemical control. PMID- 15884927 TI - Total synthesis of (-)-kendomycin exploiting a Petasis-Ferrier rearrangement/ring closing olefin metathesis synthetic strategy. AB - The total synthesis of (-)-kendomycin (1), a novel macrocyclic polyketide with antibacterial and antitumor activity, was achieved in 21 steps (longest linear sequence) exploiting an effective Petasis-Ferrier union/rearrangement tactic to construct the tetrahydropyran ring, a ring-closing metathesis to generate the macrocycle, and a biomimetic quinone-methide-lactol assembly. PMID- 15884928 TI - Alkyne haloallylation [with Pd(II)] as a core strategy for macrocycle synthesis: a total synthesis of (-)-haterumalide NA/(-)-oocydin A. AB - The rarely used haloallylation reaction, first described by Kaneda and Teranishi in 1974, employs a Pd(II) catalyst to join an alkyne with an allylic halide to produce a 1-halo-1,4-diene subunit. It is shown here that functionalized and tertiary allylic chlorides perform well as substrates in this reaction under the action of PdCl2(PhCN)2 in THF solution. When the alkyne is added slowly to the reaction mixture, the two reactants can be used in a nearly equimolar ratio. This fact means (i) that reasonably complex pairs of alkyne and allylic halide substrates are tolerated and, therefore, (ii) that an intramolecular version of the reaction is suitable as a core strategy for complex molecule construction. The latter is demonstrated in the macrocyclization of 2b to 17b, which is the central step in the total synthesis of (-)-haterumalide NA/(-)-oocydin A (1) that is reported. The final key to the completion of the synthesis was the choice of the acid-labile PMB ester of 1 as the penultimate intermediate. PMID- 15884929 TI - Alkenyl- and aryl[2-(hydroxymethyl)phenyl]dimethylsilanes: an entry to tetraorganosilicon reagents for the silicon-based cross-coupling reaction. AB - Alkenyl- and aryl[2-(hydroxymethyl)phenyl]dimethylsilanes, highly stable tetraorganosilicon reagents, are found to react with aryl and alkenyl iodides in the presence of a palladium catalyst and K2CO3 as a base, significantly milder conditions compared with those ever reported for the silicon-based cross-coupling reactions. The reaction tolerates a wide range of functional groups, including silyl protectors, and allows a gram-scale synthesis to recover and reuse the silicon residue. PMID- 15884930 TI - Stable mononuclear radical anions of heavier group 13 elements: [(tBu2MeSi)3E. ].[K+(2.2.2-Cryptand)] (E = Al, Ga). AB - The one-electron reduction of tris(di-tert-butylmethylsilyl)aluminum and -gallium with alkali metals (Li, Na, K) results in the formation of the corresponding radical anions [(tBu2MeSi)3Al*-] (3) and [(tBu2MeSi)3Ga]*- (4), which were isolated in the form of the potassium salt as extremely air- and moisture sensitive deep red crystals, representing the first isolable mononuclear radical anions of heavier group 13 elements. The molecular structures of both 3.[K+(2.2.2 cryptand)] and 4.[K+(2.2.2-cryptand)] were established by X-ray crystallography, which showed a nearly planar geometry around the radical centers. The EPR spectra of 3 and 4 showed strong characteristic signals with g-values of 2.005 for 3 and 2.015 for 4 with hyperfine coupling constants of a(27Al) = 6.2 mT for 3, a(69Ga) = 12.3 mT, and a(71Ga) = 15.7 mT for 4, corresponding to a planar geometry of the radical center. PMID- 15884931 TI - Highly sodium-selective fluoroionophore based on conformational restriction of oligoethyleneglycol-bridged biaryl boron-dipyrromethene. AB - A non-PET and non-PCT fluoroionophore has been designed and synthesized based on the combination of a biaryl boron-dipyrromethene fluorophore with an oligoethyleneglycol bridge. A specific red-shift response of absorption for NaClO4 in acetonitrile was demonstrated based on conformational restriction triggered by cation recognition at the bridge. The concentration of Na+ can be determined accurately using fluorescence due to a high extinction coefficient (epsilon = 50 000), a high fluorescence quantum yield (Phif = 0.68), and the sharpness of absorption and emission peaks of the boron-dipyrromethene derivative. PMID- 15884932 TI - A 2D solid-state NMR experiment to resolve overlapping aromatic resonances of thiophene-based nematogens. AB - In this study, we demonstrate the feasibility of resolving overlapping 13C chemical shift spectral lines of aromatic rings in a thiophene-based nematogen in the mesophase using a 2D PITANSEMA solid-state NMR method. This technique provided the information about chemical shift values as well as dipolar couplings that are used for determining the orientational order parameter. Large C-H dipolar coupling values measured for thiophene in contrast to phenyl rings suggest that the heterocyclic ring is not part of the molecular axis. Using the order parameter, we determined the orientation of C-H vectors of the thiophene ring. We believe that the 2D solid-state NMR can be extended to other types of liquid crystalline materials such as the banana-based mesogens for determining the orientational order and bent angle. PMID- 15884933 TI - Polymer electrolyte gate dielectric reveals finite windows of high conductivity in organic thin film transistors at high charge carrier densities. AB - Finite regions of high conductivity were observed in both n- and p-channel organic thin film transistors based on polycrystalline organic semiconductor films and a solution-processed, solid polymer electrolyte gate dielectric. The transition from a highly conductive state to a more insulating state with increasing gate bias may be attributed to the realization of carrier densities greater than 1014 charges/cm2 in the semiconductor film. PMID- 15884934 TI - Gold-catalyzed assembly of heterobicyclic systems. AB - We have described an efficient gold-catalyzed double cyclization of 1,5-enynes to afford a range of heterobicyclic compounds, including oxabicylclo[3.2.1]octenes, azabicyclo[3.2.1]octenes, oxaspiro[5.4]decene, azaspiro[5.4]decene, oxaspiro[5.5]undecene, oxabicyclo[4.3.0]nonene, azabicyclo[4.3.0]nonene, and oxabicyclo[4.4.0]decene. The mechanism of this reaction is proposed to involve a chemoselective gold-based alkyne activation, carbocyclization, intramolecular nucleophilic addition, followed by protodemetalation. The most notable aspect of this process is the efficient and diastereospecific interception of the reactive intermediate of the initial 6-endo-dig (or 5-endo-dig) cyclization with either oxygen- or nitrogen-based nucleophiles. PMID- 15884935 TI - Asymmetric organocatalytic epoxidation of alpha,beta-unsaturated aldehydes with hydrogen peroxide. AB - The first asymmetric organocatalytic epoxidation of alpha,beta-unsaturated aldehydes is presented. A chiral bisaryl-silyl-protected pyrrolidine acts as a very selective epoxidation organocatalyst using simple oxidation agents, such as hydrogen peroxide and tert-butyl hydroperoxide. The asymmetric epoxidation reactions proceed under environmental friendly reaction condition in, for example, water mixtures of alcohols, and the scope of the reaction is demonstrated by the formation of optically active alpha,beta-epoxy aldehydes in high yields and enantioselectivities >94% ee. Furthermore, the direct synthesis of the sex pheromone from an acaric mite by asymmetric epoxidation of citral is presented. PMID- 15884936 TI - Gold(I)-catalyzed intermolecular addition of phenols and carboxylic acids to olefins. AB - Ph3PAuOTf can catalyze efficient intermolecular addition of phenols and carboxylic acids to olefins under relatively mild conditions. PMID- 15884937 TI - CuBr-catalyzed direct indolation of tetrahydroisoquinolines via cross dehydrogenative coupling between sp3 C-H and sp2 C-H bonds. AB - A novel and efficient C-C bond formation method was developed via the cross dehydrogenative coupling (CDC) reaction of indoles and tetrahydroisoquinolines catalyzed by copper bromide in the presence of an oxidizing reagent, tert-BuOOH. The CDC reaction provides a simple and efficient catalytic method to construct indolyl tetrahydroisoquinolines via a combination of sp3 C-H bond and sp2 C-H bond followed by C-C bond formation. PMID- 15884938 TI - Serial ligand catalysis: a highly selective allylic C-H oxidation. AB - We are reporting a mild, chemo-, and highly regioselective Pd(II)-catalyzed allylic oxidation of alpha-olefins to furnish branched allylic esters that proceeds via a novel serial ligand catalysis mechanism in which two different ligands (i.e., vinyl sulfoxide 2 and BQ) interact sequentially with the metal to promote distinct steps of the catalytic cycle (i.e., C-H cleavage and pi-allyl functionalization, respectively). PMID- 15884939 TI - High performance of Rh(Phebox) catalysts in asymmetric reductive aldol reaction: high anti-selectivity. AB - Chiral rhodium(bisoxazolinylphenyl) complexes (1 mol %) efficiently catalyze the asymmetric reductive aldol reaction of aldehydes and alpha,beta-unsaturated esters at 50 degrees C for ca. 0.5-1.0 h with several hydrosilanes to give the corresponding beta-hydroxypropionates with extremely high anti-selectivity (up to 98%) and enantioselectivity (up to 96% ee). The stereochemical outcome is likely due to a chairlike cyclic transition state involving rhodium-(E)-enolate. PMID- 15884940 TI - Detection of a high-barrier conformational change in the active site of cytochrome P450cam upon binding of putidaredoxin. AB - The orientation of the substrate camphor in the active site of reduced CO-bound cytochrome P450cam (CYP101) as a function of reduced putidaredoxin (Pdxr) addition has been examined by NMR using perdeuterated CYP101 and perdeuterated Pdx as well as isotopically labeled d-camphor. This permits the 1H resonances of CYP101-bound camphor to be observed without interference from the signals of CYP101 or Pdx and confirms assignments of the methyl signals of camphor in the bound form. The Cys4Fe2S2 ferredoxin Pdx is the physiological redox partner and effector of CYP101. The addition of Pdx to the reduced CYP101-camphor-CO complex results in a conformational selection that is slow on the chemical shift time scale with spectral effects observed primarily at the 8-CH3 group of the camphor. The camphor signals are ring current shifted by the heme, and for the 9- and 10 CH3 resonances, these shifts are reasonably well predicted by ring current calculations from the crystal structure of CO-bound CYP101. However, in the absence of Pdx, the 8-CH3 resonance of CYP101-bound camphor is observed at considerably higher field than predicted. Dynamic simulations using ring current shift restraints generated a structure with low chemical shift violations in which the hydrogen bond between the camphor carbonyl oxygen and the OH of Tyr96 is lost, and an expansion of the active site takes place that permits reorientation of the camphor within the active site. PMID- 15884941 TI - Electron transfer and catalytic control by the iron-sulfur clusters in a respiratory enzyme, E. coli fumarate reductase. AB - Factors governing the efficacy of long-range electron relays in enzymes have been examined using protein film voltammetry in conjunction with site-directed mutagenesis. Investigations of the fumarate reductase from Escherichia coli, in which three Fe-S clusters relay electrons over more than 30 A, lead to the conclusion that varying the medial [4Fe-4S] cluster potential over a 100 mV range does not have a significant effect on the inherent kinetics of electron transfer to and from the active-site flavin. The results support a proposal that the reduction potential of an individual electron relay site in a multicentered enzyme is not a strong determinant of activity; instead, as deduced from the potential dependence of catalytic electron transfer, electron flow through the intramolecular relay is rapid and reversible, and even uphill steps do not limit the catalytic rate. PMID- 15884942 TI - Disodium guanosine 5'-monophosphate self-associates into nanoscale cylinders at pH 8: a combined diffusion NMR spectroscopy and dynamic light scattering study. AB - We report a combined NMR and dynamic light scattering (DLS) study on the size of supramolecular structures formed by disodium guanosine 5'-monophosphate, Na(2)(5' GMP), at pH 8. In general, two distinct types of aggregate species are present in an aqueous solution of Na(2)(5'-GMP). One type consists of stacking 5'-GMP monomers, and the other contains stacking G-quartets. Both types of aggregates can be modeled as rodlike cylinders. The cylinder diameter is 10 and 26 A for monomer aggregates and quartet aggregates, respectively. For Na(2)(5'-GMP) concentrations between 18 and 34 wt %, the cylinders formed by stacking G quartets have an average length between 8 and 30 nm, corresponding to a stack of approximately 24-87 G-quartets. These nanoscale aggregates are significantly larger than what had previously been believed for Na(2)(5'-GMP) self-association at pH 8. The length of both types of 5'-GMP aggregates was found to increase with Na(2)(5'-GMP) concentration but was insensitive to the added NaCl in solution. While the aggregate size for monomer aggregates increases with a decrease in temperature, the size of G-quartet aggregates is essentially independent of temperature. We found that the size of G-quartet aggregates is slightly larger in D(2)O than in H(2)O, whereas the size of monomer aggregates remains the same in D(2)O and in H(2)O. We observed a linear relationship between the axial ratio of the 5'-GMP cylinders and the Na(2)(5'-GMP) concentration for both types of 5'-GMP aggregates, which suggests a common stacking mechanism for monomers and G quartets. PMID- 15884943 TI - Design, synthesis, and biological evaluation of HSP90 inhibitors based on conformational analysis of radicicol and its analogues. AB - The molecular chaperone HSP90 is an attractive target for chemotherapy because its activity is required for the functional maturation of a number of oncogenes. Among the known inhibitors, radicicol, a 14-member macrolide, stands out as the most potent. A molecular dynamics/minimization of radicicol showed that there were three low energy conformers of the macrocycle. The lowest of these is the bioactive conformation observed in the cocrystal structure of radicicol with HSP90. Corresponding conformational analyses of several known analogues gave a good correlation between the bioactivity and the energy of the bioactive conformer, relative to other conformers. Based on this observation, a number of proposed analogues were analyzed for their propensity to adopt the bioactive conformation prior to synthesis. This led to the identification of pochonin D, a recently isolated secondary metabolite of Pochonia chlamydosporia, as a potential inhibitor of HSP90. Pochonin D was synthesized using polymer-bound reagents and shown to be nearly as potent an HSP90 inhibitor as radicicol. PMID- 15884944 TI - ENDOR studies of the ligation and structure of the non-heme iron site in ACC oxidase. AB - Ethylene is a plant hormone involved in all stages of growth and development, including regulation of germination, responses to environmental stress, and fruit ripening. The final step in ethylene biosynthesis, oxidation of 1 aminocyclopropane-1-carboxylic acid (ACC) to yield ethylene, is catalyzed by ACC oxidase (ACCO). In a previous EPR and ENDOR study of the EPR-active Fe(II) nitrosyl, [FeNO],(7) complex of ACCO, we demonstrated that both the amino and the carboxyl moieties of the inhibitor d,l-alanine, and the substrate ACC by analogy, coordinate to the Fe(II) ion in the Fe(II)-NO-ACC ternary complex. In this report, we use 35 GHz pulsed and CW ENDOR spectroscopy to examine the coordination of Fe by ACCO in more detail. ENDOR data for selectively (15)N labeled derivatives of substrate-free ACCO-NO (E-NO) and substrate/inhibitor bound ACCO-NO (E-NO-S) have identified two histidines as protein-derived ligands to Fe; (1,2)H and (17)O ENDOR of samples in D(2)O and H(2)(17)O solvent have confirmed the presence of water in the substrate-free Fe(II) coordination sphere (E-NO). Analysis of orientation-selective (14,15)N and (17)O ENDOR data is interpreted in terms of a structural model of the ACCO active site, both in the presence (E-NO-S) and in the absence (E-NO) of substrate. Evidence is also given that substrate binding dictates the orientation of bound O(2). PMID- 15884945 TI - Development of aliphatic alcohols as nucleophiles for palladium-catalyzed DYKAT reactions: total synthesis of (+)-hippospongic acid A. AB - The ability to use aliphatic alcohols as competent nucleophiles in the palladium catalyzed dynamic kinetic asymmetric transformation of Baylis-Hillman adducts is explored. High yield and enantioselectivity is obtained for both the kinetic transformation and dynamic kinetic transformation. The absolute stereochemistry of the products is used to explore the reactive conformation of 2-substituted pi allyl complexes with DPPBA-based chiral ligands. The utility of this method is further demonstrated in the context of a concise total synthesis of the gastrulation inhibitor (+)-hippospongic acid A. The synthesis features three palladium-catalyzed allylic alkylation reactions to introduce three different bond types: C-S, C-H, and C-O. PMID- 15884946 TI - Dynamic and structural NMR studies of cavitand-based coordination cages. AB - The interionic structure, kinetic stability, and degree of anion encapsulation of coordination cages 1 were studied by PGSE, NOE, and EXSY NMR techniques. The rate constants for the formation/dissociation processes at 296 K were obtained independently via (1)H-NOESY and (19)F-NOESY experiments giving, respectively, k(obs) = 0.30 +/- 0.04 s(-1) in CDCl(3) and k(obs) = 5.2 +/- 0.8 s(-1) in CD(3)NO(2)/CDC(13) (7.1) mixture with the proton probe, and k(obs) = 0.33 +/- 0.06 s(-1) in CDCl(3) and k(obs) = 5.0 +/- 0.8 s(-1) in CD(3)NO(2)/CDC(13) (7/1 mixture) with the (19)F probe. PGSE experiments showed that in CDCl(3) not only the encapsuled anion but also the external anions translate with the same rate as the cage. (19)F,(1)H-HOESY experiments indicated that an average of five external triflate anions are located in the equatorial sites close to the palladium moieties, while two of them approach the polar pockets formed by the alkyl chains. In a CD(3)NO(2)/CDCl(3) (7/1) mixture only one or two anions are in close proximity with the cage, while the others are solvated. In all the considered solvents (benzene, chloroform, methylene chloride, and nitromethane) the inclusion of a single unsolvated triflate anion in the cage is quantitative. (19)F,(1)H-HOESY experiments indicated that the charged guest head points toward one metal center. Therefore, while the ionic aggregation level and kinetic stability of coordination cages 1 are solvent dependent, anion encapsulation is not. PMID- 15884947 TI - Intramolecular general acid catalysis of phosphate transfer. nucleophilic attack by oxyanions on the PO3 2- group. AB - Phosphate transfer from the 8-dimethylammonium-naphthyl-1-phosphate monoanion 4m to water and to a range of nucleophiles shows general acid catalysis by the neighboring NH(+) group, through the strong intramolecular hydrogen bond. Reactivity is insensitive to the charge on the nucleophile, so that fluoride and oxyanions displace dimethylaminonaphthol from the PO3(2-) group as effectively as do amines of the same basicity. Reactivity is (predictably) relatively insensitive to the basicity of the nucleophile and to the alpha-effect. A strong intramolecular hydrogen bond is present in the product, but also in the reactant, as evidenced by major perturbations in the pK(a)'s of the phosphate and dimethylamino groups, to 3.94 and 9.31, respectively, and by ab initio calculations. Rate accelerations are of the order of 10(6)-fold despite this stabilization: the strength of the hydrogen bond is evidently significantly enhanced in the transition state. The evidence suggests that it also depends remarkably strongly on the degree of ionization of the reacting phosphate group and will be significantly reduced for the neutral PO(OH)2 group. Thus, the hydrolysis of the substrate cation 4+ shows a correspondingly greater, >10(8) fold acceleration. PMID- 15884948 TI - External electric field effects on state energy and photoexcitation dynamics of diphenylpolyenes. AB - External electric field effects on state energy and photoexcitation dynamics have been examined for para-substituted and unsubstituted all-trans-diphenylpolyenes doped in a film, based on the steady-state and picosecond time-resolved measurements of the field effects on absorption and fluorescence. The substitution dependence of the electroabsorption spectra shows that the dipole moment of the substituted stilbene in the Franck-Condon excited state becomes larger with increasing difference between the Hammet constants of the substituents. Fluorescence quantum yields of 4-(dimethylamino)-4'-nitrostilbene and 4-(dimethylamino)-4'-nitrodiphenylbutadiene are markedly reduced by an electric field, suggesting that the rates of the intramolecular charge transfer (CT) from the fluorescent state to the nonradiative CT state are accelerated by an external electric field. The magnitude of the field-induced decrease in fluorescence lifetime has been evaluated. The isomerization of the unsubstituted all-trans-diphenylpolyenes to the cis forms is shown to be a significant nonradiative pathway even in a film. Field-induced quenching of their fluorescence as well as field-induced decrease in fluorescence lifetime suggests that the trans to cis photoisomerization is enhanced by an electric field. PMID- 15884949 TI - Picosecond dynamics of nonthermalized excited states in tris(2,2 bipyridine)ruthenium(II) derivatives elucidated by high energy excitation. AB - The picosecond excited-state dynamics of several derivatives have been investigated using high photon energy excitation combined with picosecond luminescence detection. Instrument response-limited fluorescence (tau(1) approximately equal to 3-5 ps) at 500 nm was observed for all of the complexes, while longer-lived emission (tau(2) > 50 ps), similar in energy, was observed for only some of the complexes. Interestingly, the presence of tau(2) required substitution at the 4,4-positions of the bipyridine ligands and D(3) symmetry for the complex; only the 4,4-substituted homoleptic complexes exhibited tau(2). On the basis of previous assignments of the ultrafast dynamics measured for Ru(bpy)(2+)3 and Ru(dmb)(2+)3, tau(2) has been tentatively ascribed to relaxation from higher electronic or vibrational levels in the triplet manifold having slightly more triplet character than the state responsible for tau(1). However, given that the kinetics for these transition metal complexes are highly dependent on both pump and probe wavelengths and that there is considerable interest in utilizing such complexes for electron transfer in the nonergodic limit, further characterization of the state giving rise to tau(2) is warranted. PMID- 15884950 TI - A combined theoretical and experimental study of efficient and fast titanocene catalyzed 3-exo cyclizations. AB - The mechanism of titanocene mediated 3-exo cyclizations was investigated by a combined theoretical and experimental study. A gradient corrected density functional theory (DFT) method has been scaled against titanocene dichloride, the parent butenyl radical, and in bond dissociation energy (BDE) calculations. The BP86 method using density fitting, and a basis set of triple-zeta quality emerged as a highly reliable tool for studying titanocene mediated radical reactions. The computational results revealed important kinetic and thermodynamic features of cyclopropane formation. Surprisingly, the beta-titanoxy radicals, the first intermediates of our investigations, were demonstrated to possess essentially the same thermodynamic stabilization as the corresponding alkyl radicals by comparison of the calculated BDEs. In contrast to suggestions for samarium mediated reactions, the cyclization was shown to be thermodynamically favorable in agreement with earlier kinetic studies. It was established that stereoselectivity of the cyclization is governed by the stability of the intermediates and thus the trans disubstituted products are formed preferentially. The observed ratios of products are in good to excellent agreement with the DFT results. By a combination of computational and experimental results, it was also shown that for the completion of the overall cyclopropane formation the efficiency of the trapping of the cyclopropylcarbinyl radicals is decisive. PMID- 15884951 TI - Preparation of copper ion complexes of sterically congested diaryldiazomethanes having a pyridine ligand and characterization of their photoproducts. AB - To realize fairly stable high-spin polycarbenes by utilizing heterospin systems comprising 2p spins of organic radicals and 3d spins of magnetic metal ions, we prepared dianthryldiazomethanes having two pyridyl groups at the 2,2'- or 2,7 positions, that is, bis[10-(4-tert-butyl-2,6-dimethylphenyl)-2-(4-pyridyl)-9 anthryl]diazomethane (2,2'-DPy-1-N(2)) and [10-(4-tert-butyl-2,6-dimethylphenyl) 9-anthryl][(10-(4-tert-butyl-2,6-dimethylphenyl)-2,7-di(4-pyridyl)-9 anthryl]diazomethane (2,7-DPy-1-N(2)). The triplet carbene DPy-(3)1 generated by photolysis of DPy-1-N(2) was characterized by ESR and UV-vis spectroscopy in a matrix at low temperature as well as by time-resolved UV-vis in solution at room temperature. The results showed that the triplet carbene DPy-(3)1 was destabilized to some extent as opposed to the parent triplet carbene before pyridination, but it was still fairly persistent, having a half-life of more than 30 min in solution at room temperature. Photoproducts from the complex between DPy-1-N(2) and Cu(hfac)(2) were characterized in a similar manner, and the results suggested that the generated carbene centers interacted magnetically with the Cu(II) ion to form a high-spin species with significant thermal stability. The fact that no significant signals due to the isolated triplet carbene DPy-(3)1 were observed suggested that the pyridine moiety binds with Cu(hfac)(2) in a nearly quantitative manner under these cryogenic conditions. Magnetic measurements of the photoproduct using a superconducting quantum interference device (SQUID) magneto/susceptometer were performed to determine the spin state of the complex. The temperature dependence of the molar paramagnetic susceptibility indicated the presence of ferromagnetic interaction. The field dependences of magnetization for the complexes, expressed using M versus H/T plots, were analyzed in terms of the two-component Brillouin function to be S = 3.18 (F = 0.66) and S = 0.02 (F = 0.23) for the 1:1 complex of 2,7-DPy-1 and Cu(hfac)(2) and S = 2.70 (F = 0.33) and S = 0.49 (F = 0.11) for the 1:1 complex of 2,2'-DPy-1 and Cu(hfac)(2). PMID- 15884952 TI - On the electrophilicity of hydroxyl radical: a laser flash photolysis and computational study. AB - The rate coefficients for reactions of hydroxyl radical with aromatic hydrocarbons were measured in acetonitrile using a novel laser flash photolysis method. Comparison of kinetic data obtained in acetonitrile with those obtained in aqueous solution demonstrates an unexpected solvent effect on the reactivity of hydroxyl radical. In particular, reactions of hydroxyl radical with benzene were faster in water than in acetonitrile, and by a significant factor of 65. Computational studies, at the B3LYP and CBS-QB3 levels, have confirmed the rate enhancement of hydroxyl radical addition to benzene via calculation of the transition states in the presence of explicit solvent molecules as well as a continuum dielectric field. The origin of the rate enhancement lies entirely in the structures of the transition states and not in the pre-reactive complexes. The calculations reveal that the hydroxyl radical moiety becomes more anionic in the transition state and, therefore, looks more like hydroxide anion. In the transition states, solvation of the incipient hydroxide anion is more effective with water than with acetonitrile and provides the strong energetic advantage for a polar solvent capable of hydrogen bonding. At the same time, the aromatic unit looks more like the radical cation in the transition state. The commonly held view that hydroxyl radical is electrophilic in its reactions with DNA bases is, therefore, strongly dependent on the ability of the organic substrate to stabilize the resulting radical cation. PMID- 15884953 TI - Design, synthesis, structure, and gas (N2, Ar, CO2, CH4, and H2) sorption properties of porous metal-organic tetrahedral and heterocuboidal polyhedra. AB - A strategy based on assembling metal ions and organic carboxylate links has been applied for the design and synthesis of a new class of porous, truncated tetrahedral and heterocuboidal polyhedra, whose pore size and functionality can be systematically varied. The synthesis of this series of metal-organic polyhedra (MOPs) employs sulfate-capped oxygen-centered iron-carboxylate trimers, Fe3O(CO2)3(-)(SO4)3, as rigid nodes separated by linear (phenyl, biphenyl, terphenyl, and tetrahydropyrene) or trigonal (benzenetriphenyl) links to yield five highly crystalline polyhedra of general formula [NH2(CH3)2]8[Fe12O4( )(SO4)12(link)x(py)12].G (x = 6 for linear or 4 for trigonal, py = pyridine, G = guests). In this series, the size of each polyhedron has been varied from 20.0 to 28.5 A (on edge), and the corresponding pore diameter from 7.3 to 13.3 A. Gas sorption isotherms were measured for three members of this series to reveal significant uptake of gases (N2, Ar, CO2, H2, CH4) and benzene and exhibit Type I sorption behavior that is indicative of permanent porosity. The apparent surface areas for these compounds range from 387 to 480 m(2)/g. PMID- 15884954 TI - Intramolecular charge transfer in 4-aminobenzonitriles does not necessarily need the twist. AB - In electron donor/acceptor species such as 4-(dimethylamino)benzonitrile (DMABN), the excitation to the S(2) state is followed by internal conversion to the locally excited (LE) state. Dual fluorescence then becomes possible from both the LE and the twisted intramolecular charge-transfer (TICT) states. A detailed mechanism for the ICT of DMABN and 4-aminobenzonitrile (ABN) is presented in this work. The two emitting S(1) species are adiabatically linked along the amino torsion reaction coordinate. However, the S(2)/S(1) CT-LE radiationless decay occurs via an extended conical intersection "seam" that runs almost parallel to this torsional coordinate. At the lowest energy point on this conical intersection seam, the amino group is untwisted; however, the seam is accessible for a large range of torsional angles. Thus, the S(1) LE-TICT equilibration and dual fluorescence will be controlled by (a) the S(1) torsional reaction path and (b) the position along the amino group twist coordinate where the S(2)/S(1) CT-LE radiationless decay occurs. For DMABN, population of LE and TICT can occur because the two species have similar stabilities. However, in ABN, the equilibrium lies in favor of LE, as a TICT state was found at much higher energy with a low reaction barrier toward LE. This explains why dual fluorescence cannot be observed in ABN. The S(1)-->S(0) deactivation channel accessible from the LE state was also studied. PMID- 15884955 TI - A direct comparison of one- and two-component dendritic self-assembled materials: elucidating molecular recognition pathways. AB - This paper compares and contrasts, for the first time, one- and two-component gelation systems that are direct structural analogues and draws conclusions about the molecular recognition pathways that underpin fibrillar self-assembly. The new one-component systems comprise l-lysine-based dendritic headgroups covalently connected to an aliphatic diamine spacer chain via an amide bond. One-component gelators with different generations of headgroup (from first to third generation) and different length spacer chains are reported. The self-assembly of these dendrimers in toluene was elucidated using thermal measurements, circular dichroism (CD) and NMR spectroscopies, scanning electron microscopy (SEM), and small-angle X-ray scattering (SAXS). The observations are compared with previous results for the analogous two-component gelation system in which the dendritic headgroups are bound to the aliphatic spacer chain noncovalently via acid-amine interactions. The one-component system is inherently a more effective gelator, partly as a consequence of the additional covalent amide groups that provide a new hydrogen bonding molecular recognition pathway, whereas the two-component analogue relies solely on intermolecular hydrogen bond interactions between the chiral dendritic headgroups. Furthermore, because these amide groups are important in the assembly process for the one-component system, the chiral information preset in the dendritic headgroups is not always transcribed into the nanoscale assembly, whereas for the two-component system, fiber formation is always accompanied by chiral ordering because the molecular recognition pathway is completely dependent on hydrogen bond interactions between well-organized chiral dendritic headgroups. PMID- 15884956 TI - Designing PbSe nanowires and nanorings through oriented attachment of nanoparticles. AB - Single-crystal PbSe nanowires are synthesized in solution through oriented attachment of nanocrystal building blocks. Reaction temperatures of 190-250 degrees C and multicomponent surfactant mixtures result in a nearly defect-free crystal lattice and high uniformity of nanowire diameter along the entire length. The wires' dimensions are tuned by tailoring reaction conditions in a range from approximately 4 to approximately 20 nm in diameter with wire lengths up to approximately 30 microm. PbSe nanocrystals bind to each other on either {100}, {110}, or {111} faces, depending on the surfactant molecules present in the reaction solution. While PbSe nanocrystals have the centrosymmetric rocksalt lattice, they can lack central symmetry due to a noncentrosymmetric arrangement of Pb- and Se-terminated {111} facets and possess dipole driving one-dimensional oriented attachment of nanocrystals to form nanowires. In addition to straight nanowires, zigzag, helical, branched, and tapered nanowires as well as single crystal nanorings can be controllably prepared in one-pot reactions by careful adjustment of the reaction conditions. PMID- 15884957 TI - Spin-state tuning at pseudotetrahedral d(7) ions: examining the structural and magnetic phenomena of four-coordinate [BP3]CoII-X systems. AB - Electronic structure, spin-state, and geometrical relationships for a series of pseudotetrahedral Co(II) aryloxide, siloxide, arylthiolate, and silylthiolate complexes supported by the tris(phosphino)borate [BP(3)] ligands [PhBP(3)] and [PhBP(i)()(Pr)(3)] ([PhB(CH(2)PPh(2))(3)](-) and [PhB(CH(2)P(i)()Pr(2))(3)](-), respectively) are described. Standard (1)H NMR, optical, electrochemical, and solution magnetic data, in addition to low-temperature EPR and variable temperature SQUID magnetization data, are presented for the new cobalt(II) complexes [PhBP(3)]CoOSiPh(3) (2), [PhBP(3)]CoO(4-(t)()Bu-Ph) (3), [PhBP(3)]CoO(C(6)F(5)) (4), [PhBP(3)]CoSPh (5), [PhBP(3)]CoS(2,6-Me(2)-Ph) (6), [PhBP(3)]CoS(2,4,6-(i)()Pr(3)-Ph) (7), [PhBP(3)]CoS(2,4,6-(t)()Bu(3)-Ph) (8), [PhBP(3)]CoSSiPh(3) (9), [PhBP(3)]CoOSi(4-NMe(2)-Ph)(3) (10), [PhBP(3)]CoOSi(4 CF(3)-Ph)(3) (11), [PhBP(3)]CoOCPh(3) (12), [PhBP(i)()(Pr)(3)]CoOSiPh(3) (14), and [PhBP(i)()(Pr)(3)]CoSSiPh(3) (15). The low-temperature solid-state crystal structures of 2, 3, 5-10, 12, and 15 are also described. These pseudotetrahedral cobalt(II) complexes are classified as featuring one of two limiting distortions, either umbrella or off-axis. Magnetic and spectroscopic data demonstrate that both S = (1)/(2) and S = (3)/(2) ground-state electronic configurations are accessible for the umbrella distorted structure type, depending on the nature of the X-type ligand, its denticity (eta(1) versus eta(3)), and the tripodal phosphine ligand employed. Off-axis distorted complexes populate an S = (1)/(2) ground-state exclusively. For those four-coordinate complexes that populate S = (1)/(2) ground states, X-ray data show two Co-P bond distances that are invariably shorter than a third Co-P bond. The pseudotetrahedral siloxides 2, 10, and 11 are exceptional in that they display gradual spin crossover in the solid state. The diamagnetic cobalt(III) complex {[PhBP(3)]CoOSiPh(3)}{BAr(4)} ({16}{BAr(4)}) (Ar = Ph or 3,5-(CF(3))(2)-C(6)H(3)) has also been prepared and structurally characterized. Accompanying electronic structure calculations (DFT) for complexes 2, 6, and {16}(+) support the notion of a close electronic structure relationship between these four-coordinate systems and octahedral, sandwich, and half-sandwich coordination complexes. PMID- 15884958 TI - Controllable water channel gating of nanometer dimensions. AB - The dynamics of water molecules in a single-walled carbon nanotube (SWNT) under continuous deformations was studied with molecular dynamics simulations. The flux and occupancy remain almost fixed within a deformation of 2.0 A but decrease sharply for a further deformation of 0.6 A. The nanopore is an excellent on-off gate that is both effectively resistant to deformation noises and sensitive to available signals. Biological water channels are expected to share this advantage due to similar wavelike water distributions. The minimal external force required for triggering an open-close transition falls within the working range of many available experimental facilities, which provides the possibility of developing SWNT-based nanoscale devices. PMID- 15884959 TI - Cyclocarbopalladation involving an unusual 1,5-palladium vinyl to aryl shift as termination step: theoretical study of the mechanism. AB - A DFT/B3LYP model study has been carried out on the cyclocarbopalladation and on an unusual 1,5 vinyl to aryl palladium shift which are the two first steps of a cyclocarbopalladation-Stille coupling tandem reaction of various gamma bromopropargylic-1,2 diols with alkenyls or alkynyl stannanes catalyzed by Pd(PPh(3))(4). From the calculations, the active intermediates in the catalytic process appear to bear a single phosphine ligand, the palladium(II) center keeping in all cases a square-planar coordination pattern either through intramolecular binding of the triple bond or via an intramolecular Pd...C(phenyl) interaction. The computation of the various transition states and intermediates for the 1,5 vinyl to aryl palladium shift reveals that the intimate mechanism of this pathway corresponds to a one-step hydrogen transfer between the two negatively charged carbon atoms of the vinyl and phenyl groups. A two-step pathway involving a Pd(IV) intermediate is not likely to occur. This conclusion may apply to other 1,n-palladium shifts which have been experimentally observed in various organometallic transformations. PMID- 15884960 TI - Molecular manipulation of two- and three-dimensional silica nanostructures by alkoxysilylation of a layered silicate octosilicate and subsequent hydrolysis of alkoxy groups. AB - A novel methodology for constructing molecularly ordered silica nanostructures with two-dimensional (2-D) and three-dimensional (3-D) networks has been developed by using a stepwise process involving silylation of a layered silicate octosilicate with alkoxytrichlorosilanes [ROSiCl(3), R = alkyl] and subsequent reaction within the interlayer spaces. Alkoxytrichlorosilanes react almost completely with octosilicate, bridging two closest Si-OH (or -O(-)) sites on the silicate layers, to form new five-membered rings. The unreacted functional groups, Si-Cl and Si-OR, are readily hydrolyzed by the posttreatment with a water/dimethyl sulfoxide (DMSO) or water/acetone mixture, leading to the formation of two types of silicate structures. The treatment with a water/DMSO mixture produced a unique crystalline 2-D silicate framework with geminal silanol groups, whereas a water/acetone mixture induced hydrolysis and subsequent condensation between adjacent layers to form a new 3-D silicate framework. The 2 D structure is retained by the presence of DMSO molecules within the swelled interlayer spaces and is transformed to a 3-D silicate upon desorption of DMSO. The structural modeling suggests that both of the 3-D silicates contain new cagelike frameworks where solvent molecules are trapped even at high temperature (up to 380 degrees C, in the case of acetone). Both 2-D and 3-D silica structures are quite different from known layered silicates and zeolite-like materials, indicating the potential of the present approach for precise design of various silicate structures at the molecular level. PMID- 15884961 TI - Simulations on the thermal decomposition of a poly(dimethylsiloxane) polymer using the ReaxFF reactive force field. AB - To investigate the failure of the poly(dimethylsiloxane) polymer (PDMS) at high temperatures and pressures and in the presence of various additives, we have expanded the ReaxFF reactive force field to describe carbon-silicon systems. From molecular dynamics (MD) simulations using ReaxFF we find initial thermal decomposition products of PDMS to be CH(3) radical and the associated polymer radical, indicating that decomposition and subsequent cross-linking of the polymer is initiated by Si-C bond cleavage, in agreement with experimental observations. Secondary reactions involving these CH(3) radicals lead primarily to formation of methane. We studied temperature and pressure dependence of PDMS decomposition by following the rate of production of methane in the ReaxFF MD simulations. We tracked the temperature dependency of the methane production to extract Arrhenius parameters for the failure modes of PDMS. Furthermore, we found that at increased pressures the rate of PDMS decomposition drops considerably, leading to the formation of fewer CH(3) radicals and methane molecules. Finally, we studied the influence of various additives on PDMS stability. We found that the addition of water or a SiO(2) slab has no direct effect on the short-term stability of PDMS, but addition of reactive species such as ozone leads to significantly lower PDMS decomposition temperature. The addition of nitrogen monoxide does not significantly alter the degradation temperature but does retard the initial production of methane and C(2) hydrocarbons until the nitrogen monoxide is depleted. These results, and their good agreement with available experimental data, demonstrate that ReaxFF provides a useful computational tool for studying the chemical stability of polymers. PMID- 15884962 TI - Electron binding energies of aqueous alkali and halide ions: EUV photoelectron spectroscopy of liquid solutions and combined ab initio and molecular dynamics calculations. AB - Photoelectron spectroscopy combined with the liquid microjet technique enables the direct probing of the electronic structure of aqueous solutions. We report measured and calculated lowest vertical electron binding energies of aqueous alkali cations and halide anions. In some cases, ejection from deeper electronic levels of the solute could be observed. Electron binding energies of a given aqueous ion are found to be independent of the counterion and the salt concentration. The experimental results are complemented by ab initio calculations, at the MP2 and CCSD(T) level, of the ionization energies of these prototype ions in the aqueous phase. The solvent effect was accounted for in the electronic structure calculations in two ways. An explicit inclusion of discrete water molecules using a set of snapshots from an equilibrium classical molecular dynamics simulations and a fractional charge representation of solvent molecules give good results for halide ions. The electron binding energies of alkali cations computed with this approach tend to be overestimated. On the other hand, the polarizable continuum model, which strictly provides adiabatic binding energies, performs well for the alkali cations but fails for the halides. Photon energies in the experiment were in the EUV region (typically 100 eV) for which the technique is probing the top layers of the liquid sample. Hence, the reported energies of aqueous ions are closely connected with both structures and chemical reactivity at the liquid interface, for example, in atmospheric aerosol particles, as well as fundamental bulk solvation properties. PMID- 15884963 TI - An efficient fragment-based approach for predicting the ground-state energies and structures of large molecules. AB - An efficient fragment-based approach for predicting the ground-state energies and structures of large molecules at the Hartree-Fock (HF) and post-HF levels is described. The physical foundation of this approach is attributed to the "quantum locality" of the electron correlation energy and the HF total energy, which is revealed by a new energy decomposition analysis of the HF total energy proposed in this work. This approach is based on the molecular fractionation with conjugated caps (MFCC) scheme (Zhang, D. W.; Zhang, J. Z. H. J. Chem. Phys. 2003, 119, 3599), by which a macromolecule is partitioned into various capped fragments and conjugated caps formed by two adjacent caps. We find that the MFCC scheme, if corrected by the interaction between non-neighboring fragments, can be used to predict the total energy of large molecules only from energy calculations on a series of small subsystems. The approach, named as energy-corrected MFCC (EC MFCC), computationally achieves linear scaling with the molecular size. Our test calculations on a broad range of medium- and large molecules demonstrate that this approach is able to reproduce the conventional HF and second-order Moller Plesset perturbation theory (MP2) energies within a few millihartree in most cases. With the EC-MFCC optimization algorithm described in this work, we have obtained the optimized structures of long oligomers of trans-polyacetylene and BN nanotubes with up to about 400 atoms, which are beyond the reach of traditional computational methods. In addition, the EC-MFCC approach is also applied to estimate the heats of formation for a series of organic compounds. This approach provides an appealing approach alternative to the traditional additivity rules based on either bond or group contributions for the estimation of thermochemical properties. PMID- 15884964 TI - Quantum-chemical predictions of absolute standard redox potentials of diverse organic molecules and free radicals in acetonitrile. AB - A calibrated B3LYP/6-311++G(2df,2p)//B3LYP/6-31+G(d) method was found to be able to predict the gas-phase adiabatic ionization potentials of 160 structurally unrelated organic molecules with a precision of 0.14 eV. A PCM solvation model was benchmarked that could predict the pK(a)'s of 15 organic acids in acetonitrile with a precision of 1.0 pK(a) unit. Combining the above two methods, we developed a generally applicable protocol that could successfully predict the standard redox potentials of 270 structurally unrelated organic molecules in acetonitrile. The standard deviation of the predictions was 0.17 V. The study demonstrated that computational electrochemistry could become a powerful tool for the organic chemical community. It also confirmed that the continuum solvation theory could correctly predict the solvation energies of organic radicals. Finally, with the help of the newly developed protocol we were able to establish a scale of standard redox potentials for diverse types of organic free radicals for the first time. Knowledge about these redox potentials should be of great value for understanding the numerous electron-transfer reactions in organic and bioorganic chemistry. PMID- 15884965 TI - Probing charge-transfer processes in helium nanodroplets by optically selected mass spectrometry (OSMS): charge steering by long-range interactions. AB - Electron impact ionization of a helium atom in a helium nanodroplet is followed by rapid charge migration, which can ultimately result in the localization of the charge on an atomic or molecular solute. This process is studied here for the cases of hydrogen cyanide, acetylene, and cyanoacetylene in helium, using a new experimental method we call optically selected mass spectrometry (OSMS). The method combines infrared laser spectroscopy with mass spectrometry to separate the contributions to the overall droplet beam mass spectrum from the various species present under a given set of conditions. This is done by vibrationally exciting a specific species that exists in a subset of the droplets (for example, the droplets containing a single HCN molecule). The resulting helium evaporation leads to a concomitant reduction in the ionization cross sections for these droplets. This method is used to study the charge migration in helium and reveals that the probability of charge transfer to a solvated molecule does not approach unity for small droplets and depends on the identity of the solvated molecule. The experimental results are explained quantitatively by considering the effect of the electrostatic potential (between the charge and the embedded molecule) on the trajectory of the migrating charge. PMID- 15884966 TI - Infrared spectroscopy of gas-phase Cr+ coordination complexes: determination of binding sites and electronic states. AB - Infrared spectra were recorded for a series of gas-phase Cr+ complexes using infrared multiphoton dissociation (IRMPD) in a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. The functionalized aromatic ligands (acetophenone, anisole, aniline, and dimethyl aniline) offer a choice of either aromatic ring-pi or n-donor-base binding sites. Use of the FELIX free electron laser light source allowed convenient, rapid scanning of the chemically informative wavelength range from approximately 500 to 1800 cm(-1), which in many cases characterized the preferred site of metal binding, as well as the electronic spin state of the complex. Mono-complex ions, Cr+(ligand), for anisole, aniline, and dimethyl aniline and bis-complex ions, Cr+(ligand)(2), for anisole, aniline, and acetophenone were produced by ligand attachment to laser desorbed Cr+ ions in the FT-ICR cell. The photodissociation yields plotted as a function of wavelength were interpreted as approximations to the infrared absorption spectra and were compared with computed spectra of different possible geometries and spin states. Clear-cut diagnostic features in the spectra of the acetophenone, anisole, and aniline complexes showed the sites of Cr+ attachment to be the carbonyl oxygen site for acetophenone (bis-complex) and the ring-pi site for anisole and aniline (both mono- and bis-complexes). The bis-complexes of aniline and anisole are low-spin (probably doublet) states, while the mono complexes of these same ligands are high-spin (sextet) states. The dimethyl aniline complex gave a cluttered spectrum in poor agreement with calculations, which may reflect a mixture of binding-site isomers in this case. PMID- 15884967 TI - Substrate-mediated intermolecular interactions: a quantitative single molecule analysis. AB - Long-range intermolecular interactions mediated by the surface are believed to be responsible for many effects in surface science, including molecular ordering, formation of nanostructures, and aligning reactive intermediates in catalysis. Here, we use scanning tunneling microscopy to probe the weak substrate-mediated interactions in benzene overlayers on Au{111} at 4 K. Using an automated procedure to monitor single molecule motion, we are able to quantify the substrate-mediated interaction strength. We explain quantitatively both the kinetics of the benzene motion and the thermodynamics that determine the packing structures benzene adopts in this system in light of these substrate-mediated interactions. PMID- 15884971 TI - Confocal fluorescence imaging of photosensitized DNA denaturation in cell nuclei. AB - The double-stranded helical structure of DNA is maintained in part by hydrogen bonds between strands and by stacking interactions between adjacent purine and pyrimidine bases in one strand. The transition (denaturation) from a double stranded (ds) to a single-stranded (ss) form can be induced in isolated DNA or fixed cells by exposure to elevated temperatures, alkali or acids, aprotic or nonpolar solvents or some drugs. We report here that DNA denaturation can occur in situ in cell nuclei as a result of interaction between light and an intercalated dye, acridine orange or ethidium bromide. This DNA photodenaturation was probed using metachromatic properties of acridine orange and imaged by fluorescence confocal microscopy. Furthermore, an empirical kinetic model was developed to separate changes of acridine orange luminescence intensities caused by photobleaching from those that were a result of DNA denaturation. We investigated the influence of oxygen on these phenomena and propose a mechanism by which photodenaturation may occur. PMID- 15884972 TI - Synthesis of hydroxy and methoxy perylene quinones, their spectroscopic and computational characterization, and their antiviral activity. AB - Hydroxy and methoxy perylene quinones are synthesized in an attempt to isolate the essential spectroscopic and biological features of light-induced antiviral agents such as hypericin and hypocrellin. Unlike their naturally occurring counterparts, these synthetic quinones bear the carbonyl, hydroxyl, and methoxy groups in the "bay region." The hydroxy and methoxy compounds have rich absorption spectra with broad features in the visible (approximately 450-800 nm) and relatively more intense and narrow features at wavelengths < or = 350 nm. High-level ab initio quantum mechanical calculations assign the features in the absorption spectra to electronic transitions from S0 to S2 and to higher-lying electronic states. The calculations indicate that in the ground state the trans dihydroxy isomer is 12.5 kcal/mol lower in energy than the cis dihydroxy isomer and is thus the only species present. The lowest-energy trans methoxy ground state isomer and the lowest-energy cis methoxy ground state isomer are found to be degenerate. An additional cis methoxy isomer 6.3 kcal/mol higher in energy than the global minimum is assumed to contribute to the spectrum and is also considered. Finally, the synthetic compounds exhibit similar light-induced antiviral activity to each other, but significantly less than that of hypericin. PMID- 15884973 TI - South Africa--serious about biodiversity science. AB - A new government act, the creation of several centres of excellence and an injection of funding all indicate that biodiversity science is thriving in South Africa. PMID- 15884974 TI - Structure of the Mg-chelatase cofactor GUN4 reveals a novel hand-shaped fold for porphyrin binding. AB - In plants, the accumulation of the chlorophyll precursor Mg-protoporphyrin IX (Mg Proto) in the plastid regulates the expression of a number of nuclear genes with functions related to photosynthesis. Analysis of the plastid-to-nucleus signaling activity of Mg-Proto in Arabidopsis thaliana led to the discovery of GUN4, a novel porphyrin-binding protein that also dramatically enhances the activity of Mg-chelatase, the enzyme that synthesizes Mg-Proto. GUN4 may also play a role in both photoprotection and the cellular shuttling of tetrapyrroles. Here we report a 1.78-A resolution crystal structure of Synechocystis GUN4, in which the porphyrin-binding domain adopts a unique three dimensional fold with a "cupped hand" shape. Biophysical and biochemical analyses revealed the specific site of interaction between GUN4 and Mg-Proto and the energetic determinants for the GUN4.Mg-Proto interaction. Our data support a novel protective function for GUN4 in tetrapyrrole trafficking. The combined structural and energetic analyses presented herein form the physical-chemical basis for understanding GUN4 biological activity, including its role in the stimulation of Mg-chelatase activity, as well as in Mg-Proto retrograde signaling. PMID- 15884975 TI - Subversion of cellular autophagosomal machinery by RNA viruses. AB - Infection of human cells with poliovirus induces the proliferation of double membraned cytoplasmic vesicles whose surfaces are used as the sites of viral RNA replication and whose origin is unknown. Here, we show that several hallmarks of cellular autophagosomes can be identified in poliovirus-induced vesicles, including colocalization of LAMP1 and LC3, the human homolog of Saccharomyces cerevisiae Atg8p, and staining with the fluorophore monodansylcadaverine followed by fixation. Colocalization of LC3 and LAMP1 was observed early in the poliovirus replicative cycle, in cells infected with rhinoviruses 2 and 14, and in cells that express poliovirus proteins 2BC and 3A, known to be sufficient to induce double-membraned vesicles. Stimulation of autophagy increased poliovirus yield, and inhibition of the autophagosomal pathway by 3-methyladenine or by RNA interference against mRNAs that encode two different proteins known to be required for autophagy decreased poliovirus yield. We propose that, for poliovirus and rhinovirus, components of the cellular machinery of autophagosome formation are subverted to promote viral replication. Although autophagy can serve in the innate immune response to microorganisms, our findings are inconsistent with a role for the induced autophagosome-like structures in clearance of poliovirus. Instead, we argue that these double-membraned structures provide membranous supports for viral RNA replication complexes, possibly enabling the nonlytic release of cytoplasmic contents, including progeny virions, from infected cells. PMID- 15884976 TI - The neural basis of birdsong. AB - Songbirds represent an excellent model system for understanding the neural mechanisms underlying learning. PMID- 15884977 TI - Function of a fly motion-sensitive neuron matches eye movements during free flight. AB - Sensing is often implicitly assumed to be the passive acquisition of information. However, part of the sensory information is generated actively when animals move. For instance, humans shift their gaze actively in a sequence of saccades towards interesting locations in a scene. Likewise, many insects shift their gaze by saccadic turns of body and head, keeping their gaze fixed between saccades. Here we employ a novel panoramic virtual reality stimulator and show that motion computation in a blowfly visual interneuron is tuned to make efficient use of the characteristic dynamics of retinal image flow. The neuron is able to extract information about the spatial layout of the environment by utilizing intervals of stable vision resulting from the saccadic viewing strategy. The extraction is possible because the retinal image flow evoked by translation, containing information about object distances, is confined to low frequencies. This flow component can be derived from the total optic flow between saccades because the residual intersaccadic head rotations are small and encoded at higher frequencies. Information about the spatial layout of the environment can thus be extracted by the neuron in a computationally parsimonious way. These results on neuronal function based on naturalistic, behaviourally generated optic flow are in stark contrast to conclusions based on conventional visual stimuli that the neuron primarily represents a detector for yaw rotations of the animal. PMID- 15884978 TI - A three-stemmed mRNA pseudoknot in the SARS coronavirus frameshift signal. AB - A wide range of RNA viruses use programmed -1 ribosomal frameshifting for the production of viral fusion proteins. Inspection of the overlap regions between ORF1a and ORF1b of the SARS-CoV genome revealed that, similar to all coronaviruses, a programmed -1 ribosomal frameshift could be used by the virus to produce a fusion protein. Computational analyses of the frameshift signal predicted the presence of an mRNA pseudoknot containing three double-stranded RNA stem structures rather than two. Phylogenetic analyses showed the conservation of potential three-stemmed pseudoknots in the frameshift signals of all other coronaviruses in the GenBank database. Though the presence of the three-stemmed structure is supported by nuclease mapping and two-dimensional nuclear magnetic resonance studies, our findings suggest that interactions between the stem structures may result in local distortions in the A-form RNA. These distortions are particularly evident in the vicinity of predicted A-bulges in stems 2 and 3. In vitro and in vivo frameshifting assays showed that the SARS-CoV frameshift signal is functionally similar to other viral frameshift signals: it promotes efficient frameshifting in all of the standard assay systems, and it is sensitive to a drug and a genetic mutation that are known to affect frameshifting efficiency of a yeast virus. Mutagenesis studies reveal that both the specific sequences and structures of stems 2 and 3 are important for efficient frameshifting. We have identified a new RNA structural motif that is capable of promoting efficient programmed ribosomal frameshifting. The high degree of conservation of three-stemmed mRNA pseudoknot structures among the coronaviruses suggests that this presents a novel target for antiviral therapeutics. PMID- 15884979 TI - The epigenome network of excellence. AB - An initiative funded by the European Union is building a collaborative network of established and younger research groups to tackle key questions in epigenetics. PMID- 15884980 TI - Functional implications of sleep development. AB - Why do we sleep? The sleep patterns and mechanisms that occur throughout development may give us a clue. PMID- 15884981 TI - The third age of phage. AB - The third age of phage has begun with the recognition that phages may be key to the great planetary biogeochemical cycles and represent the greatest potential genetic resource in the biosphere. PMID- 15884982 TI - Developing a culturally appropriate depression prevention program: the family coping skills program. AB - Depression is a disorder that can have particularly deleterious effects on individuals from racial/ethnic minority and low-income backgrounds. Culturally appropriate prevention programs offer a way to provide accessible and effective mental health services to these underserved populations. The authors introduce the Family Coping Skills Program (FCSP), a novel depression prevention program developed specifically for low-income Latina mothers. The authors present the theoretical underpinnings of the FCSP and describe their efforts to make the program culturally appropriate and to enhance recruitment and retention of participants. Initial outcome data from an uncontrolled trial were promising and support continued development and evaluation of the FCSP and other similar programs. PMID- 15884983 TI - Ethnic matching between therapist and patient in psychotherapy: an overview of findings, together with methodological and conceptual issues. AB - This article reviews the empirical support for ethnic matching between therapist and patient in psychotherapy. The research has 3 sources: analog studies, archival studies of number of attended sessions and dropout rates, and process outcome studies of psychotherapy. Clinical trials studying ethnic matching are absent. Empirical support for ethnic matching suffers from low validity and is inconclusive, with few studies of actual psychotherapy. The research is hampered by poor conceptualization of key concepts, difficulties in forming ethnically homogeneous groups for comparisons, and an abundance of uncontrolled within-group variables. Therapist variables, for example cultural sensitivity, are rarely investigated. There is a need for large-scale psychotherapy studies with well defined key concepts in which the impacts of within-group and therapist variables are investigated. PMID- 15884984 TI - Benefits of cultural exposure and development of Korean perspective-taking ability for transracially adopted Korean children. AB - This study investigated the benefits of cultural exposure for transracially adopted (TRA) Korean children's developmental understanding of being Korean. Fifty TRA Korean children living in the United States were interviewed to assess their understanding of the implications of their Korean status for peer relations and personal identities, using a model of perspective-taking ability (PTA). Results showed cultural exposure and chronological age were significant predictors of children's PTA for being Korean. Cultural exposure had a slightly stronger relationship with PTA development for younger children than older ones. Comparison with another sample suggested that nonadopted native Korean children in the United States for an average of 25 months achieved levels of PTA at earlier ages did TRA children but that level of cultural exposure mediated these age differences. PMID- 15884985 TI - Why are hispanics at greater risk for PTSD? AB - Several studies have found that Hispanic Americans have higher rates of posttraumatic stress disorder (PTSD) than non-Hispanic Caucasian and Black Americans. The authors identified predictors of PTSD symptom severity that distinguished Hispanic police officers (n=189) from their non-Hispanic Caucasian (n=317) and Black (n=162) counterparts and modeled them to explain the elevated Hispanic risk for PTSD. The authors found that greater peritraumatic dissociation, greater wishful thinking and self-blame coping, lower social support, and greater perceived racism were important variables in explaining the elevated PTSD symptoms among Hispanics. Results are discussed in the context of Hispanic culture and may be important for prevention of mental illness in the fastest growing ethnic group in the United States. PMID- 15884986 TI - A qualitative investigation of the cultural adjustment experiences of Asian international college women. AB - This qualitative study explored the cultural adjustment experiences of 15 Asian Indian, Japanese, Korean, and Vietnamese international college women through semistructured interviews. By using consensual qualitative research methodology (C. E. Hill, B. J. Thompson, & E. N. Williams, 1997), 6 primary domains or themes related to these women's cultural adjustment experiences were identified via data analysis: their feelings and thoughts about living in the United States, perceived differences between their country of origin and the United States, their English language acquisition and use, their prejudicial or discriminatory experiences in the United States, their peer and family networks, and their strategies for coping with cultural adjustment problems. Implications of the findings for mental health practice are discussed. PMID- 15884987 TI - Hospital and after: experience of patients and carers in rural and remote north Queensland, Australia. AB - INTRODUCTION: This article reports a study of patients and carers discharged from the Townsville General Hospital into rural and remote communities in north Queensland, Australia. The findings indicate the importance of focusing on the experiences of patients and carers in attempting to understand the impact of discharge procedures. The four stories and their implications exist within a particular healthcare context that impacts disproportionately on rural patients and their carers. Economic rationalism has shaped contemporary healthcare policy in Australia, creating a system that is encouraged to conform to market principles. The costs borne by individuals, groups and communities have been increasingly privatised. Later admission and earlier discharge from hospital is now the norm. Concern about the impact of this policy context on the lives of rural and remote patients and carers prompted the study, which aimed to: (1) examine, from the patient and carer perspective, the social, economic, cultural and emotional cost of hospitalisation away from home communities; (2) identify the needs of rural and remote patients and their carers before, during and after hospitalisation a long way from home; and (3) make recommendations for improved policies and practices concerning the continuum of care: from admission planning, through hospitalisation, discharge-planning, and post-discharge support, in the context of rural and remote location. METHODS: The experiences presented highlight the depth of the challenges faced by patients and their carers who live in rural and remote communities. Both quantitative and qualitative methodologies were used to obtain insight into the complexity of patients and carers' lives. The four vignettes presented in this paper are taken from in-depth, qualitative interviews with 12 patients and 12 carers. RESULTS: The four stories described reveal the high financial and emotional costs, for patients and carers, of negotiating a healthcare system a long way from home. Challenges faced included inadequate admission planning, excessive accommodation and transport costs, and lack of post-discharge support services in home communities, as well as business failure, marital and family strain. There was an over-reliance on carers who lacked medical caring expertise, had other major commitments (family and work) and who may have had a tenuous and uncertain relationship with the patient. CONCLUSIONS: Recognition of these complex circumstances, exacerbated by rural location, during the discharge planning process should mean that attention to ensuring patients and their carers are linked to adequate support services in their communities is of the highest priority. The communication and dissemination of information to patients and carers is also vital. Information on hospital admission, travel benefits, accommodation options, care requirements post discharge are particular recommendations. To summarise, the experiences highlighted in this study suggest that patients and carers in rural and remote communities have not benefited from adequate discharge planning, and are struggling to cope in a policy context that encourages later admission, earlier discharge and over reliance on family and friends as carers. PMID- 15884988 TI - Time as a source of conflict: student nurse experiences of clinical practice in a rural setting. AB - INTRODUCTION: Conducted in rural Tasmania, this study explored the experiences of undergraduate student nurse first engagements with rural clinical practice. While the purpose of the overall study was to explore how the experience of nursing practice in a rural setting influenced the way undergraduate students shaped their professional identity, this article concentrated on the way the students came to use their time during their placement. METHODS: This study used a fusion of ethnography and hermeneutic philosophy using participant observation, field notes, interviews and reflective journaling to collect data. Data analysis took place concurrently with the data collection, which is consistent with the hermeneutic circle framework. RESULTS: Three major themes were developed and explored throughout the study: (1) navigating rural spaces; (2) time as a source of conflict; and (3) developing rural nurse identity. The theme described in this article concerns how the nursing students' assumptions regarding how time would be used during the rural placement were dislodged. This first involved a shift from regarding time as a background feature of daily life to seeing it as an explicit, foreground feature of everyday life. Second, it involved a shift away from conceptualising time as a cyclical unit of hours, instead focussing on the meaning the students came to attach to time in the rural practice setting. CONCLUSIONS: The findings shed light on how undergraduate student nurses used their time during clinical practice in a rural community. With no clinical experience, on the commencement of the rural placement they initially expected to spend their time caring for people, but realised they first needed to spend their time learning about rural nursing before they could then do the work of rural nursing. PMID- 15884989 TI - Change of place, change of pace, change of status: rural community training for junior doctors, does it influence choices of training and career? AB - INTRODUCTION: The Rural and Remote Area Placement Program (RRAPP) began in April 2000 to provide a rural community practice training term for junior doctors and to increase the opportunities for junior doctors to experience training outside the hospital setting. Recent research into the community-based training and experience for junior doctors in Australia suggests that such experience contributes to their decision-making about future training and career. METHODS: A structured national survey was undertaken of all 107 junior doctors who had participated in RRAPP prior to October 2003 and included semi-structured interviews of 54 participants from prior to October 2002. RESULTS: Data indicated that rural and community experience influenced the choice of further rural and general practice training and also provided a useful setting for junior doctors to reflect on, and confirm, future training plans. This study provided evidence of the positive influence of RRAPP on the career choices of junior doctors, with greater than 70% of participants confirming RRAPP's influence on their plans. This study also provided insight into the process of these career decisions. Decision-making was precipitated by taking junior doctors 'outside their comfort zone' of the tertiary hospital and providing a different perspective on both the present and the future. CONCLUSION: In addition to the contrast in setting and the expansion of knowledge about rural community practice, RRAPP junior doctors identified the change of place, the change of pace, and the change of status as instrumental in their decision-making about future training and careers. PMID- 15884990 TI - Community involvement in medical practitioner recruitment and retention: reflections on experience. AB - The provision of health services to rural and remote communities has been the source of much concern and debate in recent times. One aspect of this is the universal problem of insufficient medical practitioners in rural areas and the associated issues of recruitment and retention. Rural communities can play an important role in the recruitment and retention of health professionals, particularly in terms of aiding the integration of health professionals and their families into the community. Community 'involvement' is not community 'development' in the usual sense of that term. Community involvement is about engaging and facilitating active community participation and leadership in the process at hand. This article reflects on experience gained through working with rural and remote communities in Queensland, Australia, with the key purpose of facilitating active community involvement in the recruitment and retention of medical practitioners. This article raises and discusses a number of issues arising from these experiences, with particular focus on barriers and opportunities to community involvement, and working with other agencies. Communities and agencies that attempt to increase rural community involvement in health service planning, provision, recruitment and retention should consider the following. For communities: Involvement must be real--active participation; Expectations need to be achievable (short and long term); Outcomes should be sustainable; Resources and capacity should remain in the community. For agencies: Avoid creating unrealistic community expectations; Be aware of time and resource requirements and constraints; Be consistent, forthright and honest in all dealings with communities; Keep communities informed of pending policy changes; See the process through to whatever conclusion. PMID- 15884991 TI - Management systems response to improving immunization coverage in developing countries: a case study from Cambodia. AB - INTRODUCTION: In contrast to the initial success following the establishment of the National Immunization Program (NIP) in Cambodia in 1986, infant vaccination coverage rates against the six expanded program immunization diseases have not improved since 1995. In response, the NIP of the Ministry of Health has undertaken a series of institutional initiatives to address the problem of static or declining rates of coverage. The aim of this paper is to describe and assess management strategies undertaken by the NIP in Cambodia in support of improved immunization coverage. METHODS: Sources of information used in preparing this report include international literature, national coverage and surveillance data, government policy documentation, information generated by national strategic planning and health centre microplanning processes, a functional analysis of human resources, and data quality audits. RESULTS: The NIP has implemented planning, organizational development and human resource development responses to the problem of low coverage. These have included: integration of the nip strategic and operational plans into the health sector plan; strengthening of needs-based microplanning; establishment of a national monitoring and management support strategy; and the introduction of performance-based agreements between levels of government for improved immunization coverage. CONCLUSIONS: Our analysis of these findings, in particular of the international literature, suggests that NIP's responses have been appropriate, and that the development of NIP management systems and capacity will increase the likelihood for sustained immunization coverage gains within a reform environment of health system decentralization. In 2003, there are early signs that the reform processes undertaken by the NIP have resulted in improved immunization coverage in targeted areas, and this should place the national program in a stronger position to lift immunization coverage in 2004. PMID- 15884992 TI - Planning research in rural and remote areas. AB - INTRODUCTION: In order to set a regional research agenda, an interactive research workshop was planned by the joint University of Queensland and University of Southern Quennsland Centre for Rural and Remote Area Health (CRRAH), in which researchers and regional organisations would meet together to discuss and prioritise local research needs, then formulate constructive ideas and activities. METHODS: Selection of Participants: Organisations representing all key consumer, academic and health professionals within the Toowoomba, Queensland, Australia region were sent a letter inviting them to attend and to send at least one representative, resulting in a total of 75 workshop participants from 45 organisations representing 20 separate entities. The Design of the Workshop: The workshop was planned as an interactive research workshop, with a preliminary brainstorming to identify and prioritise topics, followed by facilitated small group discussions, and finally presentations to the reassembled total group. RESULTS: Forty three topics were put forward by participants during the plenary session, in which the following 12 major themes were evident: (1) health professional development and support; (2) mechanisms for identifying regional/local needs; (3) mental health; (4) health and interaction with the environment; (5) management of common conditions of which little is known; (6) post acute/aged care; (7) evidence based practice; (8) health workforce including volunteers; (9) indigenous health; (10) access to health service delivery; (11) economic impact of new programs; and (12) outcomes impact of research partnerships. Five subject areas from four of these themes were chosen for further small group discussion. A summary of the views, ideas and conclusions of each group, which were presented to a plenary session of reassembled participants over a 10-15 minute period by each group facilitators, are discussed below. Following each presentation, a 5-10 minute question session was provided after each topic. CONCLUSION: A workshop, enabling rural and remote organisations and regional researchers to meet and identify local research needs attracted strong local support. Although the final benefits of the workshop remain to be determined, a number of new collaborative research avenues are now being actively explored within the region, by a number of the participants. PMID- 15884993 TI - Cooperative knowledge-making with female and male rural doctors. AB - INTRODUCTION: It is becoming increasingly difficult to engage rural doctors in survey-based research. Rural doctors in Australia are time-poor and overworked, yet it is vital that researchers find ways to engage them because they are the holders of information that is critical to effective workforce policy and planning. AIMS: To establish a cooperative research outcome with rural General Practitioners at a time when they were subject to many competing requests for contributions to various data gathering exercises. To develop and apply a knowledge-instrument for researching the practice of female doctors. METHOD: The research project began with the intention of locating a partially hidden voice, that of female rural GPs, and bringing it to the forefront. This grew out of 6 years' work with doctors which identified that women had their own relationship with rural practice and that this was not widely recognised in the professional and policy context. Each step of the research process was negotiated with the doctors to ensure that their solutions were the basis of the work, and their language was the vehicle of investigation. Dephi rounds were used to develop content for a national survey of rural GPs. Thirty-five female rural and remote GPs contributed to 3 Delphi rounds to construct the central section of the questionnaire. The work of the expert panel contributed the unique questions at the heart of a questionnaire that was sent to 2000 rural GPs in Rural Remote and Metropolitan Areas (RRMA) 4-7. The sample was stratified by RRMA and randomised for women, with a matching sample of men to provide a control group, test whether issues identified by women are relevant to men too, and allow a gender analysis. RESULTS: Sixty-three percent of the women returned usable surveys and 54% of the men. This was a comparatively high response rate, especially for a complex, 16 page questionnaire with 79 questions, administered at a time when rural doctors had become resistant to mail surveys. Fifty-six doctors commented on the questionnaire itself, most of them appreciating the topics raised. CONCLUSION: It was possible to develop a cooperative relationship with rural doctors that resulted in high rates of participation in the research, particularly from women. What women do can be researched and included in knowledge about rural practice, and men will respond to female-designed data collection instruments. Careful attention to questions of voice, presentation, communication and purpose can assist in bringing the experience of women as well as men into the research frame. PMID- 15884994 TI - Keeping women doctors in the country. AB - Women are disproportionately poorly represented in the rural Australian medical workforce and appropriate retention policies should recognise the factors contributing to a positive rural experience for female doctors. This study describes the professional, social and family experiences of 10 female general practitioners (GPs) working in rural South Australia. Variety of work, the opportunity to provide continuity of care and being valued by the community were important factors in rural recruitment and retention. Successful retention of a female rural medical workforce also required flexible on-call and after-hours arrangements, provision of additional support and employment opportunities for non-medical spouses and partners; and provision of appropriate child care services to rural medical families. To date, there have been few initiatives specifically addressing the concerns of female rural medical practitioners. An advisory body of rural women doctors and community representatives can provide practical guidance regarding the opportunities available for rural communities to effectively support their women doctors. PMID- 15884995 TI - Epidemiology of hand dermatitis among rural nursing students in mainland China: results from a preliminary study. AB - INTRODUCTION: Although hand dermatitis (HD) is known to affect nursing students worldwide, no studies of this disease have yet been conducted in China. Similarly, rural populations are often neglected in international research. METHODS: A preliminary study of female students from both grades of a hospital based nursing school was conducted in Hebei Province, China, approximately 280 km south of Beijing. There were 27 students from the 2nd year of the course (47.4%) and 30 students from the 3rd year (52.6%). The study protocol involved an anonymous questionnaire distributed to a convenience sample of both grades from the nursing school. Statistical differences in prevalence by year of study were calculated using Fisher's exact test for discrete variables and One-way Analysis of Variance (ANOVA) for continuous variables. Statistical associations between HD and demographic items were investigated using the chi2 test. Where statistical association with HD were identified, simple logistic regression was also performed to establish a basic risk magnitude. Results were compared with similar populations of student nurses from previous studies in Australia, Japan, Denmark, the Netherlands and Germany. RESULTS: HD prevalence fell from 29.6% in the 2nd year to 13.3% in the 3rd year and averaged 21.1% across both grades. Systemic allergic disease among family members was found to be statistically associated with HD (p < 0.05). Overall, our preliminary study showed that HD probably affects Chinese nursing students at rates similar to that of their Japanese counterparts, but at a higher rate than students in other investigations from Germany, Holland and Australia. The identification of familial allergic disease as a possible risk factor for HD was also novel. CONCLUSION: The investigation was conducted as a preliminary investigation, and as such, our results need to be treated with caution. Further research is recommended to more carefully elucidate the prevalence of HD among larger groups of Chinese nursing students. PMID- 15884996 TI - Seeking quality: some experiences in South Africa. AB - Although definitions of quality in healthcare may vary, it is accepted that there are standards towards which we should be aiming. Thus quality improvement is an important part of developing rural health services. At the same time rural settings provide unique challenges to this process. The quality improvement cycle provides a tool to assist rural practitioners wishing to work towards better quality health care. The cycle starts with identifying the problems that need to be addressed and thereafter forming a team to deal with the issues identified. The team together sets standards, which provide targets appropriate to the context and towards which the service should aim. They then gather data to assess how the healthcare service is currently performing in terms of those standards. On the basis of this information, an analysis is made of the problems and their causes, which then allows the team to develop a specific plan to address the important limiting factors in the context. Implementation of the plan continues on an ongoing basis, repeating the steps as needed, with evaluation occurring as part of each cycle to assess whether quality is indeed improving. The process is described as a cycle because it needs to be ongoing, in various ways, as part of continuous quality improvement. Examples of each of the stages of the cycle are given from the South African context as illustrations of the tasks inherent in quality improvement. PMID- 15884997 TI - Oil doom and AIDS boom in the Niger Delta Region of Nigeria. AB - CONTEXT: There has been a steady rise in Nigeria's HIV/AIDS burden since the first sero-prevalence survey in 1991. Nigeria's economy is mainly dependent on oil that comes from the Niger Delta Region. The majority of the people of the Niger delta region depend on fishing and farming for livelihood. Years of oil exploration activities with frequent oil spillages have led to severe environmental degradation with resultant destruction of farmlands and aquatic flora and fauna. The situation of the people of Niger Delta is characterised by poverty, high fertility and disease. ISSUE: Oil has become, to the people of the Niger Delta region a doom that has contributed in no small measure to the AIDS boom in the region. The relationship between such a burden and poverty has been forged by years of official neglect, and the presence of foreign workers. In addition, young Nigerians are drawn to the Niger Delta region by the oil economy, searching for non-existent jobs. Within Nigeria, the states of the Niger Delta region bear the burden of the spread of HIV/AIDS, and the region's poverty amidst plenty, has a negative impact on the population. LESSONS: The article concludes that the HIV/AIDS burden of the region deserves urgent and special attention because it has far-reaching implications, not only for control efforts in Nigeria, but also for the rest of the world, as nationals of various countries find employment in its oil fields. A number of recommendations are made. PMID- 15884998 TI - Consultant supported intermediate care--a model for remote and island hospitals. AB - INTRODUCTION: Providing local consultant-delivered hospital services in remote and island communities in the United Kingdom is increasingly problematic due to difficulties with recruitment and retention of staff, statutory restrictions to hours worked by health professionals and the expectation each clinician must manage an externally defined volume of cases to maintain clinical standards. This article describes a before-and-after evaluation of a novel method of providing consultant support for acute internal medicine to an island grouping off the Scottish coast. Under the scheme, local GPs provided acute medical care of inpatients. A consultant general physician was appointed in a district general hospital on the mainland, approximately 100 miles from the island group, to provide a lead clinician role for inpatient services at the island hospital, visiting the island on a twice-monthly basis, undertaking educational sessions and developing local guidelines and care pathways for the management of individual medical conditions. In addition, two junior doctors were appointed to the island hospital to support inpatient care. METHODS: A prospective recording system for case mix was established with agreed evidence-based protocols, developed as integrated care pathways (ICP), for indicator conditions. General case mix was determined during two 6-month periods, June-November 2001 and June November 2002, before and after implementation of the new arrangements. Performance against an ICP for management of suspected cardiac chest pain was evaluated in detail, examining the process of management, clinical outcome and economics. Data from the clinical literature were used to estimate the potential health gains from observed changes in clinical practice. RESULTS: Total admissions rose by 25% in the second time period, with particular increases noted for cardiovascular, cerebrovascular disease, and cancer. Total air ambulance transfers between the islands and the mainland within these time periods increased by 31%, from 88 to 115 transfers. Recording specific details from the history and frequency of appropriate blood investigations increased and initial steps in management changed considerably after introduction of the ICP. The number of transfers to the mainland teaching hospital increased from 3/37 (8%) in 2001 to 15/56 (27%) in 2002. Based on an estimated 100 patients per year, of whom 15 would receive thrombolysis, total additional patient costs would be 64,000 pounds sterling. The annual cost of the additional resource input into the medical service was 148,000 pounds sterling. Approximately 16 adverse events would be avoided at a combined cost of 212,000 pounds sterling (148,000 pounds sterling direct costs of intervention + 64,000 pounds sterling additional treatment costs) or 13,250 pounds sterling per event avoided. This is a conservative estimate of benefit as all the direct costs of the intervention have been included. CONCLUSIONS: This study shows that appropriate standards of care can be delivered in the setting described. Costs of care increased, but the level of service provided increased concomitantly, and the health benefits were achieved at costs that compare favourably with other interventions recommended by health technology assessment groups. An estimate of notional costs involved in alternative models for the delivery of hospital medical services in a remote area suggests that costs would be similar for a three-consultant service, the present model, and a triage and transfer system. In the future, the models chosen by remote and island communities and healthcare providers are therefore likely to be determined by viability, sustainability and public acceptability rather than cost. Our study indicates that consultant supported intermediate care is a viable model. PMID- 15884999 TI - Rural Canadian community health and quality of life: testing a workbook to determine priorities and move to action. AB - BACKGROUND: Rural residents, rural community leaders, rural planners, rural health authorities and community organizations seek to understand the health, quality of life and sustainability of their communities. The aim of this article is to follow the cooperative community-based process of developing and testing a workbook to be used to assess and foster rural communities' health, quality of life and ultimately their sustainability. ISSUE: In a Social Sciences and Humanities Research Council of Canada funded, 3 phase project, entitled Determinants of Health of Rural Populations and Communities, researchers at Brandon University, University of Manitoba and Concordia University, Canada, partnered various stakeholder groups in an visionary effort to build a framework and indicators for the purpose of assisting rural communities to assess not only their health and wellbeing, but also their sustainability. CONCLUSION: In this on going project, it is anticipated that the findings related to the health, wellbeing, quality of life and sustainability of rural communities will be integral to policy development by local, provincial and federal organizations and governments well into this century. Moreover it is expected that diverse community-based organizations will be able to use the findings to take action, particularly in an intersectoral manner, the outcome of which will be an improvement of the rural resident and community health. PMID- 15885000 TI - Reframing the HIV/AIDS debate in developing countries I: setting the scene. AB - In 2001, it was estimated that 4.7 million South Africans were living with HIV/AIDS. Prevalence figures have risen steadily over the past 10 years in most African countries, and in only a few, like Uganda, does the epidemic show signs of waning. In Africa, the status of the obese has risen enormously. If you're fat, you don't have AIDS. The epidemic is fuelled by many factors. These include: a lack of basic education about HIV risk; migrant labour disrupting family stability; polygamous marriages; patriarchal practices; and a lack of basic human rights, including being able to refuse intercourse, for women. Health care systems struggle and fail to cope with overwhelming demands, and the scale of the human loss risks dehumanising carers and health workers. Doctors evolve coping strategies, as do health systems and governments. Aid from the Western world has, until recently, been tokenistic in scale. HIV/AIDS in Africa is substantially a result of the socioeconomic and political realities of the past and present, and is related to the continued exploitation of developing countries by developed nations. This article was the introductory paper in the 'HIV/AIDS in the Developing World workshop' of the 2003 WONCA World Rural Health Congress, Santiago de Compostela, Spain. The concluding 'Santiago de Compostela Statement on HIV/AIDS' was adopted at the congress, and is offered here as a suggested way forward. PMID- 15885001 TI - Reframing the HIV/AIDS debate in developing countries II: case stories of Nigerian rural people living with HIV/AIDS. AB - Since it was first officially reported in Nigeria in 1984, HIV/AIDS has continued to spread in the country to the extent that today it constitutes a major problem. The two case stories of Nigerian rural people living with HIV/AIDS presented in this article were selected from 200 such stories that were compiled against the backdrop of the impact of scarcity of resources on care and support for People Living With HIV/AIDS from Nigerian rural communities. PMID- 15885002 TI - Reframing the HIV/AIDS debate in developing countries III: an effective, equitable response. AB - Developing countries need to balance resources for treatment and prevention. In Southern Africa, only 100,000 out of 4.1 million people who need HIV/AIDS anti retroviral therapy (ART) are able to access it. The drop in the price of ART has led to opportunities to increase the numbers receiving treatment, but problems remain. Increasing health service focus on HIV might poach staff and resource from other important programs like TB, malaria or child health. It depends on good organisation and laboratory support. It may medicalise the epidemic and distract attention from the need for education and prevention. There is now good evidence that preventive strategies, including STD treatment, improved practices of blood transfusion and needle use, use of drugs to prevent mother child transmission, voluntary counselling and testing, increased condom availability and behaviour change are very effective in reducing spread. It is obvious that both treatment and prevention strategies are necessary. International aid is still inadequate. The European Union spends 50 billion dollars on agricultural subsidies, but donates only 140 million dollars for HIV in Africa. As funding increases, it is vital that it is well used and reaches the people who need it most. PMID- 15885003 TI - Reframing the HIV/AIDS debate in developing countries IV: does ethics have anything to offer? AB - Dealing with HIV/AIDS is one of the major ethical challenges facing the world today. It is suggested that an expanded discourse on ethics, divided into three levels, can help give a fuller understanding of all aspects of the HIV/AIDS pandemic. The levels are: (1) micro level (doctor-patient relationship); (2) meso level (civic and public health ethics); and (3) macro level (ethics of international relationships). At the micro level, the four principles of respect for autonomy, beneficence, non-maleficence and justice apply to HIV patients, as to any other. However, the overwhelming demand for medical care, and the lack of doctor availability in developing countries seriously limits their application. At the meso level, the Tavistock principles give a framework for health systems. The principles are: rights to health and health care; balancing resources among competing needs; comprehensiveness; cooperation among patients, clinicians and managers; focus on improvement, safety and openness. In this context, rights are respected by not discriminating on the basis of sex, geography, tribe or race. A balance has to be struck between treatment and prevention. Comprehensiveness means not ignoring palliative care and health improvement strategies. Cooperation requires 'the reciprocity and interdependence that characterise community'. The remaining principles are self-explanatory, but frequently ignored in health planning. At a macro level, there is a need for ethical discourse about issues like increasing inequality between rich and poor countries; the use of economic levers by developed countries to the disadvantage of developing countries; the international debt crisis; the tiny health care spend (US5-10 dollars per capita per annum) in Africa; and other problems like refugee and migrant labour movements. These factors fuel global instability and the HIV/AIDS pandemic, as well as contributing to the threat of terrorism and environmental degradation. We need to look at how the values of Western democracy can be revised to address these problems. For example, scientific knowledge should be made available to all who can benefit from it; individualism should be put into the context of the common good; and free market forces need to be modified to reflect the fact that we live in a world that is increasingly interdependent. PMID- 15885004 TI - General practitioners' perceptions of after hours primary medical care services: a Toowoomba, Queensland, Australia study. AB - INTRODUCTION: This article reports on a project, undertaken in 2002 in the regional city of Toowoomba, Queensland, Australia, that investigated the viability of establishing an after-hours primary medical care (AHPMC) service. OBJECTIVES: To ascertain GPs' perceptions of the adequacy of AHPMC services in Toowoomba. DESIGN: Thirty GPs were randomly selected to participate in face-to face interviews using a semi-structured interview tool. SETTING: Toowoomba, Australia is the largest inland non-capital city in Australia. It is located approximately 130 km west of Brisbane, the State capital city and is a referral centre for patients from the rural and remote communities of south-west Queensland. PARTICIPANTS: 15 male and 15 female GPs. RESULTS: While the majority of participants believed the current provision of AHPMC in Toowoomba was adequate, they stated that the provision of AHPMC services was onerous and, given a choice, they would prefer to refer all patients seeking care between 2200 and 0800 hours to an Emergency Department (ED). Similar to GPs who work in rural and remote areas of Australia, they believed that AHPMC work was poorly remunerated, had an adverse effect on their lifestyle and could endanger their personal and their patient's safety. CONCLUSION: The findings of this study confirm previous studies into the perceptions of GPs to the provision of AHPMC in a regional city. Additionally, while the GPs in Toowoomba have the options of referring after hours patients to an ED and being part of a large GP after hours cooperative, their opinions on after-hours work did not differ significantly from those expressed by GPs working in rural and remote areas of Australia. The GPs in this study, given the option, would prefer not to undertake an AHMPC service provision between 2200 and 0800 and many had chosen not to do so, instead directing their patients at this time to one of the two EDs located in Toowoomba. PMID- 15885005 TI - Delivery of the clinical components of the Newcastle University medical course in a multidisciplinary academic unit in Tamworth. AB - The University Department of Rural Health (UDRH), northern New South Wales, Australia, was established in late 2001 by the University of Newcastle in Tamworth, New South Wales (NSW). The UDRH is part of the Commonwealth Department of Health and Ageing's Health Workforce Undergraduate Initiative and was the eighth UDRH to be established in Australia. The UDRH northern NSW delivers medical education as part of a multidisciplinary academic unit. At the end of 2003, six medical students graduated having completed all of their clinical training based in Tamworth. Over 60% of the local medical community were teachers in the program. These students studied with final year dietetic, occupational therapy and radiography students co-located in Tamworth during 2002 and 2003. In 2004, ten new graduates from the program have chosen to commence employment in the New England Area Health Service, including two medical students completing their internship in 2004. This represents the first time that the full clinical curriculum of an Australian medical school has been delivered entirely in a single rural setting. It demonstrates a new way of funding medical education which embraces local clinicians as 'faculty' and provides a way for regional centres to develop their own future medical workforce. PMID- 15885006 TI - Memories of being a rural doctor's son. PMID- 15885007 TI - Doctors' perspectives on the viability of rural practice. AB - INTRODUCTION: Private practitioners play a vital role in meeting the health needs of rural communities. However, the prospect of operating a private practice business in rural Australia seems to be increasingly unattractive, because many communities are forced to recruit salaried or overseas-trained doctors. This study focuses on rural practices as businesses whose viability influences their attractiveness for the recruitment and retention of practitioners. The specific objectives are to ascertain which factors contribute to or threaten practice viability in rural areas, and whether they vary according to the degree of rurality or geographical remoteness. METHODS: This study is based on data collected from a national study into the viability of rural general practice undertaken jointly by the Rural Doctors Association of Australia and Monash University School of Rural Health Bendigo. The Rural Remote and Metropolitan Area (RRMA) classification was used as the indicator of rurality. The study surveyed all general practitioners practising in rural or remote regions of Australia (RRMAs 3 to 7). Only practitioners with some financial interest in the practice were selected for this analysis. Free-text responses to the two questions 'What are the key factors contributing to the viability of your practice?' and 'What factors would put the viability of your practice at risk?' were analysed using qualitative content analysis. Factors were derived iteratively through higher level aggregation of responses. Chi-square tests were used to make comparisons across the RRMA categories. RESULTS: The national survey achieved a response rate of 35% of the entire population of GPs practising in RRMA 3 to 7 regions. Of these, 1050 respondents were relevant to this analysis. Seven major factors were identified by practitioners as the main contributors to practice viability. 'Practice characteristics' was nominated by 59% of respondents, followed by 'Income' (31%), 'Personal circumstances', 'Workforce' and 'Community characteristics' (all approximately 23%), 'GP activities and workload' (16%) and 'Professional support' (12%). Eight main factors were identified by practitioners as threats to viability. 'Workforce' was nominated by 57% of respondents, followed by 'Financial' (44%), 'Medico-legal' (33%), 'Administration-political' (16%), 'Community characteristics' (15%), 'GP-practice characteristics' and 'Personal circumstances' (10%) and 'Family circumstances' (3%). Across RRMA 3 to 5 the order of the percentage of respondents identifying each factor was generally consistent, with significant differences in the magnitude of the percentages for three contributing factors and four risk factors. While respondent numbers in RRMA 6 and 7 communities were low, significance testing did reveal differences between them and the rural communities on two contributing and one risk factor. CONCLUSIONS: Practice viability is a major factor affecting the attractiveness of rural and remote practice for intending and existing GPs. Initiatives designed to contribute to viability will not be successful unless measures are also adopted to address perceived threats. This study highlights the systemic nature of the factors which contribute to and threaten practice viability. Although a primary component of practice viability is economic, with income from consultations being critical, the importance of the interrelationships between the main viability factors should not be underestimated. Clearly a multifaceted systemic response is required to overcome problems associated with rural workforce recruitment of future and burnout of current rural GPs. PMID- 15885008 TI - Always one doctor away from a crisis! PMID- 15885009 TI - Quality use of medicines in the rural ambulant elderly: a pilot study. AB - INTRODUCTION: Australia has a rapidly ageing population, especially in rural areas, and strategies to address medicines and the elderly are particularly relevant. The aims of this 18 month study, therefore, were to: (1) determine the influence of a medication review on the quality of life of elderly ambulatory patients managed by a general practitioner; and (2) assess the impact of the medication review process on health outcomes such as medication-related hospital admissions in ambulant elderly patients actively managed by their GP. METHODS: The study was conducted within the area serviced by the rural Riverina Division of General Practice, New South Wales, Australia. Patients were identified by clinical audit, and recruited to the study if they met the inclusion criteria of being: older than 65 years, ambulant, living independently and on five or more medications. The study sample consisted of 402 participants (156 men, 38.8%; 246 women, 61.2%). Fifty-eight participants withdrew from the study for a variety of reasons. A two-group (intervention, control) pre- and post-intervention randomized study design was utilized. Quality of life was assessed using SF-36. The medication history and clinical details of the 202 study participants were reviewed by the project pharmacist and their GP. Medication changes were suggested to patients by their GP and follow-up SF36 and review of hospitalisation episodes were conducted after 6 months. RESULTS: 3382 medications were identified as being taken, an average of 8.4 medications per patient. After the initial medication review, the study pharmacist suggested an alteration in dose, form or frequency for 687 medications in the intervention group. The GPs recommended an alteration in 243 of patient medications. Of the entire study population (n = 402), only two participants' admission to hospital was specifically attributed to medication-related issues. There were no significant differences between the quality of life assessments for the combined groups; however, the intervention group recorded significantly higher scores in two of the nine dimensions measured: vitality (p 0.009) and mental health (p 0.0001), at the post-intervention assessment. CONCLUSION: While the intervention did not reduce hospitalisation episodes and only led to a modest improvement in quality of life, the development of a mutually acceptable form of face-to-face pharmacist/GP medication review, identification of potentially serious adverse drug reactions, identification of previously unreported complementary medicine use, and enhanced GP awareness of the risks of polypharmacy were positive outcomes of the study. PMID- 15885010 TI - Prevalence of musculoskeletal conditions, associated pain and disability and the barriers to managing these conditions in a rural, Australian Aboriginal community. AB - INTRODUCTION: Internationally, musculoskeletal conditions are a major morbidity issue for Indigenous populations and the social and economic burden imposed by musculoskeletal complaints is significant. However, little is known about the prevalence and associated pain and impairment of musculoskeletal conditions among rural Indigenous Australians. METHODS: The study was conducted between January 2001 and July 2002. DESIGN: A cross-sectional research design was used. SAMPLE: Participants included 189 Indigenous members of the community, 80 of whom were randomly selected and 109 were recruited using a convenience sample. The sample included 87 males (46%) and 102 females (53%). Participants' mean age was 44 years (+/-14.8). MEASURES: The main outcome measures were sites of current pain, self-reported levels of pain, limitations to activities of daily living, and barriers to managing these conditions. PROCEDURE: Following a screening survey participants underwent a clinical examination conducted by musculoskeletal health professional trained in standardised, clinical assessment procedures. RESULTS: Lower back pain, followed by neck, head and shoulder pain were the most common conditions. Approximately 57% of participants suffered from 2 to 4 musculoskeletal conditions. The most commonly endured level of pain was 'high'. There were no significant differences between male and female participants in terms of reported levels of pain. The level of pain reported was relatively high compared with the level of associated limitation of activities of daily living. A majority of participants had suffered from their principal condition for 7 weeks or more, indicating high levels of chronicity in the community. CONCLUSION: The majority of people living in this large rural, Indigenous community have learnt to live with chronic levels of pain affecting multiple anatomical sites. Strategies such as community musculoskeletal health promotion and appropriately trained community health workers can assist the community manage this chronic burden of disability. There is scope for further study into the musculoskeletal health of both rural and urban Indigenous populations. PMID- 15885011 TI - Lateral epicondylalgia: a problem for rural workers. AB - Lateral Epicondylalgia (LE) (tennis elbow) is a problematic condition for workers in labour intensive industries. The economies of many rural communities are founded on industries such as grape growing, meat and fish processing, fruit processing and winemaking. Workers in those industries are at increased risk of developing work-related upper limb mechanical disorders, including LE. This article reviews current understanding of the aetiology and management of this common condition as an aid to rural health professionals managing this presentation. A variety of treatment options exist for the management of LE with the literature divided on which approach is most effective. However, there is evidence that exercise is a key component of a management strategy. In addition to the current practice of prescribing exercises for the wrist extensor muscles, recent research suggests that appropriate activation of the stabilising muscles of the shoulder and cervical spine also needs to be considered by the rural practitioner. PMID- 15885012 TI - Capacity building in rural health research: a Canadian perspective. AB - INTRODUCTION: The need to build capacity in rural health research is recognized in Canada. During its formative stage, the Canadian Rural Health Research Society (established in 2002), performed a nation-wide survey of the research teaching and training opportunities available to students interested in rural and remote health issues. Intended as a cross-sectional 'snapshot,' and to provide base-line data, the survey involved a comprehensive list of educational programs in health and science disciplines at all Canadian Universities offering graduate-level programs. The present project report gives an overview of the results, documenting the current strengths, but also the noticeable gaps that exist. METHODS: Data were collected via an Internet survey, developed according to Dillman's email survey design principles. Although multiple contacts are recommended, due to time constraints only two were made. The instrument included both open- and closed-ended questions designed to determine institutional commitments to rural health research training generally, as well as specific information about course offerings, infrastructure supports for students, financial assistance for studying specific topics, and future plans. Health was defined broadly and included a number of disciplines beyond those usually considered under the health rubric. Individuals in administrative positions for 462 programs in 24 distinct disciplines at 56 institutions were sent the survey. Responses were received from 120 programs at 40 universities. Due to budget constraints the instrument was not translated into French. Descriptive statistics were used to analyse the data, combined with a thematic analysis of written comments. Ethical approval was obtained from the Lakehead University before the study was conducted. RESULTS: Despite the fact that one-third of Canadians live in rural, remote and northern areas, there is a paucity of research that addresses rural health issues. Moreover, Canadian universities have very limited rural health offerings in their curricula. There are few formal courses available on rural health topics generally or rural health research specifically. Although students can take rural-related independent studies or reading courses, very few actually take advantage of the option. Of all disciplines, nursing faculty are most likely to pursue rural health research and to offer rural health courses. Moreover, nursing faculty most often indicate an intention to expand their rural health offerings. In the social sciences, geography and environmental science programs, relatively few students choose to study rural health. Similarly, in medicine, the number of students exploring rural health issues is small. There is some specific financial assistance available to students who are interested in rural health research and, as well, they have access to general pools of funding. CONCLUSION: The survey results demonstrate the limitations of current educational programs. However, they also indicate areas of potential growth and show widespread interest in increasing the offerings available at universities across Canada. The article concludes by giving an overview of the mandate and initiatives being taken by the new research society to augment training, and to enhance student participation. In addition, it notes the positive developments linked to a new Strategic Plan by the Canadian Institutes of Health Research (CIHR) to enhance rural health research in Canada. PMID- 15885013 TI - Evaluation of nine pilot obstetric ultrasound education workshops for Australian rural and remote doctors. AB - INTRODUCTION: This article describes the evaluation of nine pilot obstetric ultrasound education workshops for Australian rural and remote doctors developed in response to an educational needs assessment that showed a large unmet need in this area. METHOD: Data from pre- and post-knowledge tests were analysed in SPSS. Data from the follow-up sonographer assessment visits were analysed in an EXCEL spreadsheet. RESULTS: In all, 141 doctors attended one of the nine pilot obstetric ultrasound workshops and reported an increase in their confidence in obstetric ultrasound. The overall workshop pre- and post-knowledge test mean improved from a score of 13.9 to 15.5 (p = 0.001) and the RANZCOG pre- and post knowledge test mean improved from 9.56 to 15.12 (p = <0.001). Data from the follow-up competency assessment visits showed doctors were competent in some areas but required further practice in others. CONCLUSION: An intensive educational intervention can successfully address the pre-determined needs of rural and remote doctors. However, this intervention should be followed by ongoing practice-based education and assessment. PMID- 15885014 TI - Canadian and American self-treatment of pain: a comparison study. AB - INTRODUCTION: Adults frequently rely on self-treatment modalities to relieve pain that exceeds everyday kinds of pain, such as minor headaches and toothaches. Examples of self-treatment modalities include doctor-prescribed analgesics, non prescribed over-the counter medications, herbal substances and treatments, and non-drug treatments such as heat, cold and exercise. Self-treatment is often associated with adverse affects related to the improper use of self-treatment substances and the adverse interactions they may produce when combined with other prescribed or non-prescription treatments for pain control. Many adults also use a variety of self-treatment modalities without informing their health care providers. OBJECTIVES: To explore the occurrence of pain and identify pain self treatment modalities used by members (n = 105) of rural communities from two eastern Canadian provinces. Results of this study were compared with a rural American cohort study in order to explore similarities/differences in patterns of self-treatment of pain between the two countries. METHODS: This descriptive exploratory study was conducted using a survey method. The design followed that used in a US study by Vallerand, Fouladbakhsh and Templin. Investigators used self-report questionnaires to identify pain self-treatment modalities, pain intensity ratings, pain interference, and the percentage of pain relief in a convenience sample of 105 participants recruited from two Canadian rural communities. Differences in mean scores between Canadian and US data were determined through t-tests. Difference between Canadian and US pain self treatment modalities were determined using chi2 tests for significance. RESULTS: Canadians reported choosing significantly more non-pharmacological self-treatment modalities of pain control such as heat, cold, exercise/stretching, and massage than did their US counterparts (chi2 = 7.6, p = .006). US participants reported significantly higher percentages of pharmacologic modalities than Canadian participants, ie prescription medications (chi2 = 4.8, p = .03), and over-the counter medications (chi2 = 8.14, p = .004). There was no significant difference between the two countries in the number of herbal supplements taken for pain relief (chi2 = 2.47, p = .12). Canadian participants reported having significantly less pain relief from their current self-treatment regimen than US participants, (t = 13.77, p = .00). In addition, 33% of Canadian participants and 20% of US participants had not informed their primary care practitioner of their self-treatment choices. CONCLUSIONS: This study demonstrates that pain is a common experience for many North American individuals living in rural communities. Comparison of results between Canadian and US cohort studies indicate that rural Canadians may benefit from increasing their knowledge about self-treatment options of pain control. Findings showed that rural Canadians choose more non-pharmacologic self-treatment modalities and have less pain relief than rural US participants by their self-treatment choices. In addition, a significant number of Canadian and US participants had not informed their primary care provider of their self-treatment practices. Community healthcare agencies may need to improve the dissemination of information on how to combine both pharmacologic and non-pharmacologic modalities into self-treatment regimens in order to facilitate more effective pain control for some rural communities. Further study is indicated to examine how the differences found in self-treatment practices between Canadian and US cohort studies relate to the differences between Canadian and US culture and healthcare payment systems. PMID- 15885015 TI - Preliminary guidelines for the implementation of Community Based Rehabilitation (CBR) approaches in rural, remote and Indigenous communities in Australia. AB - A forum of health professionals was held in Brisbane, Queensland, Australia, 30 31 August 2003, to discuss the relevance and potential of the Community Based Rehabilitation (CBR) model to rural, remote and Indigenous communities in Australia. The forum identified principles and guidelines for the development of CBR, which are presented here as a focus point for future discussion and action by people with disabilities, rural community members, Indigenous people, policy makers and health professionals. Forum members noted that while considerable strengths were evident in the CBR model, it has yet to make a significant impact on the service system in Australia. While recognising that the Australian context is quite different from many countries in which CBR has traditionally been implemented, they suggested that it may have particular application to remote, rural and Indigenous communities. To facilitate the principles of CBR in these communities, the forum called for recognition of the need for greater community involvement in disability services, the need to develop appropriate training frameworks, and the need to redirect resources to such community models. In keeping with the CBR philosophy, forum members noted that if the model is to be implemented effectively, substantial consumer and community involvement will be instrumental in future steps. PMID- 15885016 TI - A virtual clinic: telemetric assessment and monitoring for rural and remote areas. AB - This article reports the establishment of a pilot 'virtual clinic' in a rural region of Victoria, Australia. Using low-cost videophones that work across ordinary phone lines, together with off-the-shelf (mostly automatic) clinical tools, local volunteers have been trained to mediate a virtual consultation between simulated patients and local GPs. This system has the potential to save long trips into town by such patients since the traditional 'home visit' is not feasible, as well as to provide regular home monitoring for those with chronic conditions. This in turn should impact favourably on ambulance deployment, sometimes enabling patients to avoid going to hospital or allowing them to come home sooner than otherwise would be the case, and generally to offer a sense of medical security to those living in isolated regions. PMID- 15885017 TI - 'What makes it really good, makes it really bad.' An exploration of early student experience in the first cohort of the Rural Clinical School in the University of Western Australia. AB - INTRODUCTION: The Australian Commonwealth Department of Health and Ageing has implemented the Government's Regional Health Strategy. This strategy provides funding to universities for the establishment of Rural Clinical Schools (RCS) on a national basis. The strategy aims to secure a rural education and training network to increase the availability and viability of rural health services in the long term. The University of Western Australia set up the RCS in 2002 with the objective of setting up a full 5th year medical course in remote communities (RRAMA [Rural Remote and Metropolitan Area Classification] 4-7) for the 2003 academic year. There were 21 students in five areas: Kalgoorlie/Esperance (9 students), Broome (4 students), Port Hedland (3 students) and Geraldton (5 students). These students covered the 5th year curriculum with internal assessment and final examinations, in the same manner as city students. Only the delivery was different, according to geographical location. METHODS: Structured questionnaires using open-ended questions were distributed to students on two occasions. At the sixth month, semi-structured interviews were held with each student. The interviews were transcribed and a thematic analysis of the data was undertaken. Constant comparison of data was undertaken, themes identified and relationships among the themes clarified. RESULTS: In general, students were very happy with the teaching and learning opportunities they had during the first half of the year. However the initial themes of curriculum content, curriculum delivery, and assessment, were eclipsed by an overarching theme of anxiety and its management. The issue of student anxiety was addressed during the analysis. A number of factors were identified which ameliorated student anxiety or contributed to increased anxiety. From this evaluation a number of contributory factors to such student anxiety were identified. The investigators became more cognisant of the impact of group dynamics and of the need to structure the 'unstructured' environment of rural and remote medicine. In this way, students focus on only a few learning tasks at a time. They complete each topic with at least one other student so they can share the experience. The key role of each site coordinator also became apparent. The site coordinator should know the curriculum thoroughly and transmit this information to other teachers and preceptors at their site. It was also found desirable that the RCS was clear as to which assessment processes were flexible and which were 'fixed'. The medical school must clarify which curriculum content is essential, which is desirable and which is additional. Issues of workload must be monitored, and good work practices must be encouraged and supported. It was found that the high level of commitment to learning lead to the potential for burnout, generating the student comment: 'What makes the RCS really, really good makes it really bad...' CONCLUSION: Setting up an innovative program is always a major task, but setting up five different offices with four centers of learning separated by thousands of kilometers has not been undertaken, apparently, anywhere else in the world. It has been a 'fast uphill journey' that has been subject to evolving change as the RCS has adapted to conditions not expected from an academic point of view. Key contributory factors to student anxiety were identified and organizational strategies were implemented immediately, where possible, to reduce such anxiety. These insights were also used in the preparation for, and implementation of, the 2004 curriculum. PMID- 15885018 TI - Working together for a sustainable urgent care system: a case study from south eastern Australia. AB - CONTEXT: A small, isolated community in the south east of Australia, Malacoota, had a long-standing concern about the adequacy of their emergency medical systems. There was no hospital, the local medical practitioners were under stress and their ambulance services were limited. Following an approach through the local Division of General Practice in August 2002, the School of Rural Health at Monash University was invited to assist. ISSUES: A policy development toolkit was used to improve the rural urgent care systems through engagement with community members. The process involved community consultation, a meeting of key stakeholders, and the formation of a representative Steering Committee to oversee the local management of the project. Project officers worked with a university facilitator and other stakeholders to implement the Transforming Rural Urgent Care Systems (TrUCs) process from August 2002 to June 2003. A proposal of recommendations was put to the Victorian State Minister of Health and this was accompanied by a degree of political action. The submission raised the issues of poor interstate communications, ambulance staffing, support for medical practitioners, facilities for the stabilisation of patients, and access to air ambulance services. Funding was obtained for the implementation of a community paramedic model. Ambulance service communications systems improved and an innovative model of ambulance service delivery for isolated communities was implemented. LESSONS: A number of lessons have been identified, including the crucial role of the project officers, and communication within the community and among specific stakeholders. The approach used could be adopted in other rural locations hoping to improve their emergency health services. PMID- 15885019 TI - Rural GPs' ratings of initiatives designed to improve rural medical workforce recruitment and retention. AB - INTRODUCTION: Since the early 1990s, Australian governments have recognised the problems of rural medical workforce recruitment and retention and have implemented a range of programs and incentives designed to improve the supply of, and access to, doctors in rural and remote areas. Some incentives involve differential payments according to degree of rurality or remoteness. Since these programs involve considerable costs to governments, some assessment of their impact on recruitment or retention is warranted. The objective of this study is to examine the effectiveness of different recruitment and retention incentives from the perspective of the rural GP. Doctors practising in rural and remote communities were, therefore, asked to rank the relative importance of different interventions in terms of their impact on recruitment to and retention of GPs in their communities. METHODS: Six possible interventions were selected to cover the major objectives underpinning rural workforce programs most relevant to doctors currently in rural practice. Respondents assigned a rank to indicate the relative importance of each item with respect to each of the two questions: "What would help most to attract more GPs to this community?" and "What would help most to retain GPs in this community?" The data were collected as part of a national study into the viability of rural general practice undertaken jointly by the Rural Doctors Association of Australia and Monash University School of Rural Health Bendigo. The Rural, Remote and Metropolitan Areas (RRMA) classification was used as the rurality indicator. Analysis involved the calculation of mean ranks for each item. Item means were then ranked to indicate most to least important items in total, and within each RRMA category. RESULTS: Thirty-five percent of all GPs practising in rural and remote Australia responded to the national survey, representing 53% of all practices in those areas. Of these, 1050 doctors who nominated themselves as a principal, partner or associate in their practice were eligible for inclusion in this analysis. The results showed a high degree of agreement in the responses to both questions, with the possible interventions being ranked in the same overall order. 'Better remuneration for Medicare consultations' and 'Improved after-hours and on-call arrangements' were ranked as the most important interventions for both attracting and retaining GPs, whereas 'Better education and professional support activity' and 'Improved availability of allied health professional services' were ranked as least important of the options presented. 'Better locum availability' and 'Capital funding to improve practice infrastructure/enable GPs to set up practice' ranked in between. Results within each RRMA category were very similar to the overall rankings, confirming the significance of the improved remuneration and workload arrangements, regardless of geographical location. CONCLUSIONS: Ensuring professional support, workforce supply, income and infrastructure support are all relevant to the recruitment and retention of GPs in rural and remote areas. However, from the perspective of GPs practising in such areas, specific initiatives that increase the core income of rural practices, and which address those medical workforce supply issues which impact most on workload, are considered those which are most likely to assist in the recruitment and retention of GPs to Australia's rural and remote communities. PMID- 15885020 TI - Effect of rurality on patients' satisfaction with out of hours care provided by a family doctor cooperative. AB - INTRODUCTION: Reacting to demand and supply pressures, European healthcare systems are undergoing significant structural changes to the organisation and delivery of out of hours care. Such pressures are of particular concern to rural practice. Although patient satisfaction with out of hours care has been extensively studied, the effect of rurality on satisfaction levels has not, to our knowledge, been previously examined. OBJECTIVE: To investigate whether rurality has an influence on patient satisfaction with out of hours care provided by a family doctor co-operative. METHODS: All patients contacting the service over a designated 24-day period were forwarded a postal questionnaire. Patients' satisfaction was measured using a version of the McKinley questionnaire, and rurality, by subjective patient assessment, distance from treatment centre or previous rota cover. RESULTS: The response rate was 55% (531/966). Overall satisfaction levels were high with 88% of patients rating the service as either good or excellent. 47.8% of respondents perceived themselves as living in a town, 14.6% as living in a village, and 37.6% as living in the countryside. Perceived rurality, distance from treatment centre or previous rota cover did not significantly affect satisfaction levels. CONCLUSIONS: Family doctor co operatives have significantly altered the way out of hours care is delivered. Patients from rural areas are equally satisfied with the provision of out of hours care by co-operatives, as urban patients. Extension of co-operatives to rural areas need not be constrained by concerns regarding decreased patient satisfaction. PMID- 15885022 TI - Medical schools must contribute to rural health care in South America. PMID- 15885021 TI - Teamwork in rural practice. PMID- 15885023 TI - Planning and implementing the first stage of an oral health program for the Pika Wiya Health Service Incorporated Aboriginal community in Port Augusta, South Australia. AB - The oral health of the Indigenous community in South Australia's mid-north has been a concern for some years. There has been a history of under-utilisation of available dental services by the local community. This is in part due to the services not meeting their cultural and holistic health care needs. The Indigenous community resolved to establish a culturally sensitive dental service within the Aboriginal Health Service already operating in Port Augusta in South Australia's mid-north. To achieve this, a partnership between Pika Wiya Health Service Incorporated, the South Australian Dental Service, the University of Adelaide Dental School and the South Australian Centre for Rural and Remote Health was formed. The aim of the project partners was to establish a culturally sensitive, quality dental service that caters to the needs of the Indigenous community serviced by Pika Wiya Health Service Inc. This article describes the process of planning and implementing the first stage of this project. PMID- 15885024 TI - A database to record, track and report health student rural placements. AB - The Spencer Gulf Rural Health School (SGRHS), South Australia, is funded by the Australian Commonwealth Government to deliver health education in the rural setting. The SGRHS required a database to record, track and report on student rural placements to satisfy Commonwealth reporting requirements, and for internal academic and administration staff use. Staff in widely separate rural locations needed to be able to access the database. A web-based relational database was created using Microsoft Access. The student rural placement database has been successfully utilised as the primary tool to record and track student placements in the SGRHS for 2 years, and has generated data for eight Commonwealth reports in this time. Future database developments include student accessible sections. With few alterations the database could be utilised by other Australian Rural Clinical Schools and University Departments of Rural Health. PMID- 15885025 TI - Adolescent health: a rural community's approach. AB - INTRODUCTION: Significant health problems encountered in adulthood often have their roots in health behaviours initiated during adolescence. In order to reverse this trend, school and health personnel, as well as parents and other community members working with high school students, need to be aware of the health-related beliefs and choices that guide the behaviours of teenagers. Although a wide variety of research has been conducted on this topic among urban adolescents, less is known about the health beliefs and behaviors of adolescents residing in rural areas, particularly in Canada. In general, rural Canadians are less healthy than their urban counterparts. Building on the knowledge and understanding of their own community, key stakeholders were invited to engage in the design and implementation of a participatory action research project aimed at understanding and improving the health of rural adolescents. METHODS: A group of parents, teachers, students, school administrators and public health nurses engaged in a participatory action research project to better understand determinants of the health of rural adolescents at a high school in Western Canada. Group members developed and administered a health survey to 288 students from a small rural high school, in an effort to identify areas of concern and interest regarding health practices and beliefs of rural adolescents, and to take action on these identified concerns. RESULTS: Results indicated some interesting but potentially worrying trends in this population. For example, while frequent involvement in a physical activity was noted by 75.9% of participants, close to half of the females (48%) described their body image as 'a little overweight' or 'definitely overweight', and approximately 25.8% of respondents noted that they skipped meals most of the time. Differences between the genders were apparent in several categories. For example, more girls smoked (16.2%) than boys (12.3%), and more males (55.0%) than females (41%) had tried illegal drugs. Participants indicated awareness of other health-compromising behaviours, including unsafe driving habits and high stress levels, and acknowledged several steps they wanted to take to improve their health, as well as the barriers to taking those steps. Students identified improved nutrition, stress reduction, and increased levels of physical activity as particular important health goals. Students also recommended ways in which information and support could be provided within the school environment to enable them to achieve their health-related goals. Several activities developed in collaboration with students have incorporated the recommendations, and have spawned other activities in response to the ongoing identification of new concerns. CONCLUSIONS: The process of including the rural community in the identification of health assets and needs from the perspective of students -- as well as the planning and implementation of appropriate strategies to address those needs -- demonstrates the strengths inherent within a small rural population. Community members' awareness of the need to create a healthy environment for youth is reflected in their willingness to participate in activities leading to improved health. Greater awareness of the health needs of rural adolescents, and of the influence of gender in some aspects of health behaviors, will help researchers to explore ways in which the unique culture of rural communities can be harnessed to help shape health-focused interventions. PMID- 15885026 TI - Factors that influence students in choosing rural nursing practice: a pilot study. AB - INTRODUCTION: Nursing shortages continue globally and are especially critical in rural and remote communities. Attracting nurses to work in less populated regions presents challenges that differ from those in urban areas. METHODS: This pilot study focused on self-identified factors of nursing students who expressed an interest in rural practice post-graduation. The sample included students from the USA and Canada, who were enrolled in graduate and under graduate programs of nursing, and were attending an international rural nursing conference. RESULTS: Findings from the pencil and paper short answer survey found those who have life experiences and connections in small communities are more likely to choose this setting. Post-graduation employment preference was reinforced by ongoing exposure to rural theory and practice settings in their programs of study. Nursing scholars may find this study useful to further examine students' employment preferences, and to develop targeted strategies to better prepare those having an interest in rural practice. CONCLUSION: Evidence based findings are critically needed to recruit and retain nurses to address critical nursing shortages in rural regions in North America and globally. PMID- 15885027 TI - Rural proofing for health: a commentary. AB - CONTEXT: Health services policies in the UK have traditionally been developed as a 'one size fits all' approach usually based on urban models. This approach can be problematic for primary care agencies involved in the delivery of services in rural and remote areas, due to difficulties with access and also the differences in the nature of urban and rural communities. The Countryside Agency in England has developed a rural proofing framework which will assess the impact that new policies may have in rural areas. This process is currently being applied across all government departments. This article sets out the context of the National Health Service (NHS) in the UK and describes the process of Rural Proofing for Health and how it can be used to improve equity of access to care for people living in rural and remote communities in Britain and across the international healthcare community. ISSUES: There has been a considerable increase in investment in the NHS in recent years with consequent improvements such as reductions in waiting times for treatment. Despite tangible improvements there are still inequalities in health and inequities in healthcare provision, which include parts of the population who live in rural areas. The Rural Proofing for Health project has been developed and carried out by the Institute of Rural Health as a methodology to help Primary Care Organisations implement policies and methods of service delivery that take into account the needs of people living in rural and remote areas. A toolkit has been developed that adopts a solutions based approach to healthcare delivery in rural areas and which will help people living in rural areas to have equity of access to care with their counterparts living in urban Britain. Rural proofing policies before implementation ensures that the possible impacts of the policy are measured and so that adjustments can be made to ensure that the service will be appropriate for rural communities, and that it can be delivered effectively and efficiently in a rural context. LESSONS: It is important that development of new healthcare policies includes a process that ensures such policies are rurally sensitive. Involving stakeholders at a strategic level ensures that the initiative becomes embedded into policy-making within each department. Rural proofing is a systematic methodology that will help policy makers take account of health needs in rural areas and will hold public bodies accountable to their rural communities. PMID- 15885028 TI - The positive impact of rural medical schools on rural intern choices. PMID- 15885030 TI - Metal ion modulation of cystinyl aminopeptidase. AB - Cystinyl aminopeptidase has one Zn2+-binding motif and is a member of the M1 aminopeptidase family. Ion modulation of its catalytic activity was studied in membranes of CHO-K1 cells (Chinese-hamster ovary K1 cells) using L-leucine-p nitroanilide as substrate. The planar bidentate chelators 1,10-phenanthroline and 2,2'-bipyridine inhibited the activity in a concentration-dependent manner with Hill slopes of 3.32+/-1.78 and 2.10+/-0.26 respectively. The acetic acid containing chelators EDTA, EGTA and DTPA (diethylenetriamine-N,N,N',N'',N''-penta acetic acid) weakly affected the activity, but they increased the potency of the planar chelators up to a limit, at which Hill slopes became close to unity. Moreover, competition between 1,10-phenanthroline and the substrate only took place in the presence of EDTA. These findings are compatible with a model in which the bidentate chelators inhibit enzyme activity by decreasing the free Zn2+ concentration. By removing a modulatory ion from an allosteric site at the enzyme, the acetic acid-containing chelators facilitate the direct interaction between the bidentate chelators and the catalytic Zn2+. The inhibitory effect of EDTA plus 1,10-phenanthroline could be completely reversed by Zn2+. Ca2+ and Mg2+ increased the potency of Zn2+ for this process. This is expected if they interact with the modulatory site to decrease the sensitivity of the enzyme towards 1,10 phenanthroline. Conversely, the bidendate chelators increased the high-affinity [125I]angiotensin IV binding to the membranes and this was potentiated by the acetic acid-containing chelators. These findings support the concept that high affinity [125I]angiotensin IV binding, previously referred to as 'AT4 receptor binding', only occurs for the cystinyl aminopeptidase apoenzyme. PMID- 15885031 TI - Modification of blood cell PrP epitope exposure during prion disease. AB - PrPC [normal cellular PrP (prion-related protein)] is a glycosylphosphatidylinositol-linked cell-surface glycoprotein that is expressed primarily by cells of the central and peripheral nervous system and the lymphoreticular system. During prion disease, PrPC undergoes structural modification to PrPSc (abnormal disease-specific conformation of PrP). The appearance of prion infectivity and PrPSc within different peripheral lymphoid tissue sites during natural scrapie infection in sheep is suggestive of haematogenic dissemination. For this to occur, blood cells may harbour or carry disease-associated PrP and in doing so present altered conformations of PrP on their cell-surface. In the present study, we show that changes in PrP epitope expression, or accessibility, can be detected on peripheral blood mononuclear cells during the course of experimental scrapie in susceptible sheep. Peripheral blood mononuclear cells isolated from VRQ homozygous lambs inoculated orally with scrapie were probed with either N- or C-terminal-specific anti-PrP monoclonal antibodies and analysed by flow cytometry. During the progression of scrapie, significant alterations were seen in the exposure of particular cell-surface PrP epitopes. These modifications included increased accessibility to N-terminal regions of the PrP molecule, to the region between beta-strand-2 and residue 171, and to the C-terminal region of helix-3. Increased accessibility in the globular C-terminal domain of PrP occurred in the vicinity of tyrosine dimers, which are believed to have increased solvent exposure in disease-associated PrP. We suggest that the alterations in anti-PrP monoclonal antibody recognition of cell-surface PrP on blood cells from scrapie-infected sheep are indicative of structural changes within this molecule that may be relevant to prion disease. PMID- 15885032 TI - Role of the vitamin D receptor in FGF23 action on phosphate metabolism. AB - FGF23 (fibroblast growth factor 23) is a novel phosphaturic factor that influences vitamin D metabolism and renal re-absorption of Pi. The goal of the present study was to characterize the role of the VDR (vitamin D receptor) in FGF23 action using VDR(-/-) (VDR null) mice. Injection of FGF23M (naked DNA encoding the R179Q mutant of human FGF23) into VDR(-/-) and wildtype VDR(+/+) mice resulted in an elevation in serum FGF23 levels, but had no effect on serum calcium or parathyroid hormone levels. In contrast, injection of FGF23M resulted in significant decreases in serum Pi levels, renal Na/Pi co-transport activity and type II transporter protein levels in both groups when compared with controls injected with mock vector or with FGFWT (naked DNA encoding wild-type human FGF23). Injection of FGF23M resulted in a decrease in 25-hydroxyvitamin D 1a hydroxylase mRNA levels in VDR(-/-) and VDR(+/+) mice, while 25-hydroxyvitamin D 24-hydroxylase mRNA levels were significantly increased in FGF23M-treated animals compared with mock vector control- or FGF23WT-treated animals. The degree of 24 hydroxylase induction by FGF23M was dependent on the VDR, since FGF23M significantly reduced the levels of serum 1,25(OH)2D3 [1,25-hydroxyvitamin D3] in VDR(+/+) mice, but not in VDR(-/-) mice. We conclude that FGF23 reduces renal Pi transport and 25-hydroxyvitamin D 1a-hydroxylase levels by a mechanism that is independent of the VDR. In contrast, the induction of 25-hydroxyvitamin D 24 hydroxylase and the reduction of serum 1,25(OH)2D3 levels induced by FGF23 are dependent on the VDR. PMID- 15885033 TI - Passive and dynamic joint stabilization in the normal and anterior cruciate ligament-deficient knee. PMID- 15885034 TI - Passive and dynamic translation in the knee is not influenced by knee exercises in healthy individuals. AB - No evidence exists that repeated isolated quadriceps contractions lead to increased sagittal plane tibial translation. The purpose was to analyze passive and dynamic tibial translation before, during and after a specific exercise session, including cycling and a maximum number of knee extensions and heel raises, in healthy individuals. Passive and dynamic sagittal tibial translation was measured on 18 healthy individuals (nine men and nine women) before, during and after a specific exercise session with heavy load, including cycling and maximum number of knee extensions and heel-raises. Sagittal tibial translation during the Lachman test, maximal isometric quadriceps contraction, one-legged squat and gait was registered with the CA-4000 electrogoniometer. The electromyographic (EMG) activity of m. vastus medialis and lateralis, m. gastrocnemius and mm. hamstrings was registered. There was no difference in passive or dynamic sagittal tibial translation during or after the exercise session. No difference could be detected in EMG activity during squat after compared with before the exercise session. In conclusion, the knee exercises did not influence the amount of translation in healthy individuals. The findings indicate that isolated strength training of quadriceps may be included in anterior cruciate ligament rehabilitation programs, even if further specific studies on injured individuals most be performed. PMID- 15885035 TI - Sagittal tibial translation during exercises in the anterior cruciate ligament deficient knee. AB - The aim of this study was to describe and compare the sagittal tibial translation and electromyographic activity of muscles v. medialis and lateralis, gastrocnemius and hamstrings, during common rehabilitation exercises, in patients with anterior cruciate ligament (ACL) deficiency and non-injured controls. Sagittal tibial translation was registered with the CA-4000 electrogoniometer, in 12 patients and 17 controls, during Lachman test (static translation) and five exercises (dynamic translation). The exercises were grouped according to muscle work and joint compression (active extension, heel raises, cycling, one-legged squat and chair squat). The non-weight-bearing exercise with isolated muscle work (active extension) produced a large amount of tibial translation. During weight bearing, the total anterior-posterior tibial motion was halved compared to the non-weight-bearing exercises, but tibia was anterior positioned. Heel raising resulted in equal translation as the one-legged squat and chair squat, whereas cycling produced the smallest amount of tibial translation. The subjects utilized different amounts of their individual joint play (static translation) during the exercises, which may explain why there are no correlation between static translation and the patients functional outcome. These results enhance understanding on tibial translation during activity and gives indications on which exercises can be used early after ACL injury and reconstruction. PMID- 15885036 TI - Permanent disabilities in the displaced muscle from rupture of the long head tendon of the biceps. AB - Patients with a displaced muscle belly because of rupture of the long head biceps tendon were investigated for local pain and other disabilities, together with strength and endurance loss. Eleven patients (median age 59 years, minimum follow up 6 months) were included, and minimum follow-up was 6 months. Magnetic resonance imaging (MRI) of both upper arms allowed investigation of muscle atrophy and evaluation of any other degenerative signs in the displaced muscle. All patients reported pain or disability locally in the displaced muscle in certain situations, and strength and endurance were reduced by 25%. MRI revealed the displaced muscle to be unreduced in size and with no signs of degeneration. Generally, operative reattachment of the displaced muscle is not advocated in middle-aged or older patients. In order to elucidate this subject, we present a retrospective consecutive series of patients with considerable disabilities in the displaced muscle belly independent of shoulder disabilities. PMID- 15885037 TI - Relationship between radiographic abnormalities of lumbar spine and incidence of low back pain in high school rugby players: a prospective study. AB - The purpose of this study was to examine the relationship between prevalent radiographic abnormalities of the lumbar spine and the incidence of low back pain in high school rugby players. Three hundred and twenty-seven incoming high school rugby players (15-16 years of age) were recruited between 1986 and 1994. All of them underwent athletic physical examination before participation in high school athletics. The relationship between prevalent radiographic abnormalities of the lumbar spine in the pre-participation athletic physical examination and the incidence of low back pain during a 1-year period after the start of participation in athletics was analyzed. Radiographic abnormalities assessed were spondylolysis, disc space narrowing, spinal instability, Schmorl's node, balloon disc, and spina bifida occulta. The frequency of respective radiographic abnormalities was 15.6%, 13.1%, 32.7%, 14.1%, 11.6%, and 30.6%. Two hundred and forty-three players (74.3%) had at least one of these radiographic abnormalities. The incidence of low back pain was 44.0% in players with no such radiographic abnormalities and 41.2% in those with at least one radiographic abnormality. The incidence of low back pain in players with the respective radiographic abnormalities was 72.5%, 46.5%, 46.7%, 39.1%, 34.2%, and 41.0%, and only players with spondylolysis had a significantly higher incidence of low back pain than those with no radiographic abnormalities. This study shows that a radiographic abnormality, specifically spondylolysis, is a significant radiological risk factor for low back pain in high school rugby players. PMID- 15885038 TI - Evaluation of general joint laxity, shoulder laxity and mobility in competitive swimmers during growth and in normal controls. AB - The aim of the study was to evaluate differences between competitive swimmers and a reference group of school children concerning general joint laxity, laxity of the glenohumeral joint and range of motion in the shoulder. Materials and methods. Competitive swimmers (n = 120) were compared with references consisting of age and gender matched school children (n = 1277). General joint laxity was evaluated with the Beighton score. Anterior glenohumeral laxity was assessed according to the drawer test, and inferior glenohumeral laxity according to the sulcus test. Shoulder rotation was measured with a goniometer. RESULTS: Male swimmers of both age groups showed a higher degree of general joint laxity compared with the reference group while 9-year-old female swimmers alone had a lower degree of general joint laxity compared with references. No significant difference concerning shoulder laxity was noticed between groups. There was a decreased internal rotation in male and female swimmers as compared with the reference group. External rotation was reduced in female swimmers as compared with the female references. The same result was observed in male swimmers, but only at the age of 12 years. CLINICAL CONSEQUENCE: Competitive swimming in children seems to lead to a decreased range of motion with regard to shoulder rotation. However, the reason for this is still unclear and further investigations are needed. PMID- 15885039 TI - Post-exercise palpation of pulse rates: its applicability to habitual exercisers. AB - Despite the increased popularity of heart rate (HR) monitors, endurance-trained adults as well as habitual exercisers often use pulse rate palpation to periodically monitor exercise intensity. However, due to the rapid recovery of HR following exercise bouts, post-exercise palpation of pulse rates may underestimate exercise HR. To test this hypothesis, we studied 20 young physically active adults performing two sets of exercise for 5 min at 70% and 85% of maximal HR on the treadmill; one with carotid and another with radial pulse count. Post-exercise palpation of pulse rate was lower (P < 0.01) than the actual HR during exercise, underestimating exercise HR by 20-27 bpm (beats per min). Even when ECG tracings of HR were analyzed immediately after exercise (0-15 s), a significant underestimation of exercise HR (7-9 bpm) still persisted (P < 0.05). Following exercise, pulse rate obtained by carotid palpation at both intensities and radial palpation at the lower intensity was no different from the corresponding HR measured with ECG. In the radial artery trial at the higher exercise intensity, pulse rate following exercise was lower (10 bpm; P < 0.05) than ECG-derived HR. Arterial stiffness, which is closely associated with arterial baroreflex sensitivity, was not significantly related to the changes in HR with carotid palpation. We concluded that post-exercise pulse palpations may not be appropriate as an indicator of exercise intensity in habitual exercisers. PMID- 15885040 TI - The influence of intermittent altitude exposure to 4100 m on exercise capacity and blood variables. AB - This study was performed to investigate the effects of intermittent hypoxic exposure on blood and exercise parameters. Eight sea level residents were exposed to 2 h daily stimulus to 4100 m altitude in a hypobaric chamber for a total of 14 days. Exercise performance was evaluated at sea level before and after the hypoxic stimulation. Blood samples were obtained before, during, and at time points up to 14 days after the hypoxic exposure. No changes were observed in haemoglobin, haematocrit, reticulocytes, serum transferrin receptors, or EPO levels in the blood. Submaximal cycle (150 W) ergometer exercise corresponded to a oxygen uptake of 1.9+/-0.1 and 1.9+/-0.1 L min(-1) before and after the intermittent altitude exposure, respectively. At maximal exercise the workloads attained were 343+/-17 and 354+/-27 W before and after the exposure, with corresponding oxygen uptakes of 4.0+/-0.2 and 4.2+/-0.2 L min(-1). It is concluded that intermittent hypoxic exposure to 4100 m altitude for 2 h daily and a total of 14 days does not affect exercise capacity. PMID- 15885041 TI - Predicting dropout in male youth soccer using the theory of planned behavior. AB - This investigation prospectively predicted dropout among young soccer players, using the theory of planned behavior (TPB). First, behavioral beliefs required to develop a TPB-questionnaire were elicited from 53 male soccer players, aged 13-15 years. Second, at the beginning of the soccer season, 354 different male soccer players aged 13-15 years completed this questionnaire, thereby assessing direct dimensions (intention, attitude, subjective norm, perceived control) and indirect dimensions (attitudinal, normative and control beliefs) derived from TPB. Nine months later--upon termination of the soccer season--these players were classified into 323 perserverers and 31 dropouts, with TPB being applied prospectively to predict these two groups. For both direct and indirect dimensions, between-group comparisons revealed significant differences in favor of the perseverers. Discriminant analyses revealed five measures (intention, attitude, subjective norm, a normative belief, and a control belief), which enabled a 22.1% a priori dropout prediction when used within a suitable equation. In conclusion, TPB may have a promising application to prospectively discriminate dropouts from perseverers, providing a potential predictive a priori classification model for sport participation. PMID- 15885043 TI - Seronegative myasthenia gravis: disease severity and prognosis. AB - Around 10-20% of myasthenia gravis (MG) patients do not have acetylcholine receptor (AChR) antibodies (seronegative), of whom some have antibodies to a membrane-linked muscle specific kinase (MuSK). To examine MG severity and long term prognosis in seronegative MG compared with seropositive MG, and to look specifically at anti-AChR antibody negative and anti-MuSK antibody negative patients. Seventeen consecutive seronegative non-thymomatous MG patients and 34 age and sex matched contemporary seropositive non-thymomatous MG controls were included in a retrospective follow-up study for a total period of 40 years. Clinical criteria were assessed each year, and muscle antibodies were assayed. There was no difference in MG severity between seronegative and seropositive MG. However, when thymectomized patients were excluded from the study at the year of thymectomy, seropositive MG patients had more severe course than seronegative (P < 0.001). One seropositive patient died from MG related respiratory insufficiency. The need for thymectomy in seronegative MG was lower than in seropositive MG. None of the seronegative patients had MuSK antibodies. This study shows that the presence of AChR antibodies in MG patients correlates with a more severe MG. With proper treatment, especially early thymectomy for seropositive MG, the outcome and long-term prognosis is good in patients with and without AChR antibodies. PMID- 15885044 TI - Preserved painting creativity in an artist with Alzheimer's disease. AB - Creativity in any of its forms, either visual, musical, literary or performing arts, may be conceived as a cognitive capability, and should be actively explored in relation to patients with Alzheimer disease and related dementias, even when other cognitive functions do not allow us to even communicate with them. We are reporting the case of a talented artist with the diagnosis of early onset Alzheimer disease (AD) with progressive cognitive impairment but with preservation of her creativity until very late in the course of the disease. PMID- 15885045 TI - Twenty-four-month comparison of immunomodulatory treatments - a retrospective open label study in 308 RRMS patients treated with beta interferons or glatiramer acetate (Copaxone). AB - A total of 308 patients with relapsing remitting multiple sclerosis (RRMS) documented in a clinical database have been analysed for comparison of treatment effects of first line immunomodulating therapies. During follow up for 24 months the reduction of relapse rates was evaluated in a retrospective and open-label clinical study by comparing the efficacy of interferon (INF)beta-1a i.m. (Avonex), INFbeta-1b s.c. (Betaferon), INFbeta-1a 22 microg s.c. (Rebif 22) and glatiramer acetate (GA; Copaxone) in patients with RRMS (enhanced disability status score (EDSS) < 3.5) who have been treated for at least 6 months. Compared with baseline, relapse rate was reduced to a comparable extend after 6 month treatment with all regimen. For all drugs the effect on the relapse rate was sustained over 24 months.. There was no superiority of one of the INFbeta preparations concerning reduction of relapse rate after 12 and 24 months, however reduction was significantly higher for patients treated with GA compared with all beta interferons (-0.71, P < 0.001). In addition, the discontinuation rate within 24 months was significantly lower for GA. Despite some limitations of the study design, the results provide helpful clinical information regarding the efficacy of immunomodulatory therapies in early stages of RRMS patients in a clinical setting. PMID- 15885046 TI - A pilot double-blind placebo-controlled trial of low-dose pramipexole in sleep related eating disorder. AB - Sleep-related eating disorder (SRED) is characterized by recurrent arousals from sleep associated with compulsive ingestion of food. No controlled therapeutic trials are available for SRED. We assessed the safety, tolerability and efficacy of pramipexole, a dopamine D3-receptor agonist, in the treatment of SRED. Eleven consecutive patients with SRED in the absence of concurrent daytime eating disorders underwent actigraphic recording and subjective sleep diary evaluation for a week before and every week for 2 weeks of treatment with pramipexole 0.18 0.36 mg or placebo, administered in a double-blind crossover randomized sequence. The primary outcomes of the trial were actigraphic measures of night sleep parameters (sleep efficiency, motor activity mean and median, number and duration of wake episodes), secondary outcomes were the number of good sleep nights/weekly, number and duration of nocturnal awakenings/night, and visual analogue preference score. Pramipexole was well tolerated without any patient withdrawing from the study. Pramipexole reduced night-time activity median (P = 0.02) and increased the number of nights of good sleep/week (P = 0.02). All other measurements remained unaffected. Pramipexole at low doses was well tolerated, improving some measures of sleep quality and reducing median night activity in SRED. Further studies with higher dosages and for longer time-periods are warranted. PMID- 15885047 TI - The mechanism of ischemic stroke in patients with dolichoectatic basilar artery. AB - Basilar artery dolichoectesia (BD) may cause brainstem ischemia by multiple mechanisms, including thrombosis, embolism, occlusion of deep penetrating arteries. The objective of this study was to determine and characterize clinical, imaging findings and hemodynamic mechanisms in patients with cerebrovascular event associated with BD and compare these data with those for patients with BD who did not have stroke. We studied 29 consecutive stroke, two transient ischemic attack (TIA) patients with BD who have been admitted to our stroke unit. We sought the diameter of ectasia, height of the bifurcation, lateral displacement, shape deformities, and blood flow velocity of the basilar artery (BA) by transcranial Doppler. Imaging and hemodynamic findings were compared with those found in a group of 18 patients without stroke or TIA. The main infarct localization was pons, eight (28%) with restricted single lesion, 10 (32%) with multiple lesions involving thalamus, midbrain, posterior cerebral artery (PCA) territory. Patients with BD were more probably to have had stroke fitting a clinical and imaging patterns of multiple infarcts than those with restricted infarct in territories supplied by branches of the BA (60% vs. 40%). Hypertension and atherosclerotic changes of the posterior circulation were more frequent in patients with stroke than those without (P = 0.004 and P = 0.028, respectively), whilst the incidence of other vascular risk factors were not significantly different in two groups. Patients with stroke/TIA had more often low blood flow velocity but not significant in the BA when compared with those for BD patients without cerebrovascular event (71% vs. 39%; P = 0.1). Reduced blood flow velocity in the BA was correlated significantly with distal lesions involving thalamus, midbrain and PCA territory rather than those located in the territory supplied by branches of the BA (P = 0.02). In conclusion, it seems probably that BD may cause vertebrobasilar system ischemia by multiple mechanisms, especially reduced blood flow in the BA and atheromatous changes in the vertebrobasilar system may precipitate thromboembolic stroke. PMID- 15885048 TI - The influence of alcohol and time on the S-100B levels of patients with minor head injury. AB - The present study aimed at evaluating the S-100B serum level's reaction to (i) alcohol consumption and (ii) time elapsing between head injury and blood sampling. Nineteen patients with minor head trauma and with at least one of the following symptoms: amnesia, transient loss of consciousness or severe headache, were included in the study. Immediately after arrival venous blood samples were drawn for determination of alcohol concentration and S-100B level. Four hours later a new blood sample was taken for repeat analysis. Twenty-one healthy volunteers drank a moderate amount of alcohol. Blood samples were taken just before alcohol intake and 4 h later. Patients - After 4 h the mean S-100B level had fallen from 0.26 to 0.21 ng/ml (P < 0.01), i.e. a mean decrease of 0.05 ng/ml, 95% confidence limits: 0.02-0.09 ng/ml. The alcohol concentrations also decreased significantly from 2.00 +/- 0.27 per thousand to 1.31 +/- 0.20 per thousand, P < 0.001, mean difference = 0.69 per thousand, 95% confidence limits: 0.27-1.11 per thousand. No difference was found between the S-100B levels of patients whose serum did contain alcohol and the levels of those whose serum did not. Volunteers - The serum alcohol level reached a mean value of 0.81 per thousand +/- 0.09 per thousand. The mean S-100B level rose from 0.077 +/- 0.02 ng/ml before alcohol intake to 0.103 +/- 0.06 ng/ml, 4 h later (P < 0.01). These data indicate that the time that elapses between trauma and blood test has an effect on the S-100B level. The same goes for the drinking of even a very moderate amount of alcohol. The relative importance of these two sources of error remains to be determined. PMID- 15885049 TI - Absence of focal neurological involvement in tick-borne relapsing fever in northern Tanzania. AB - The aim of the study was to chart incidence and clinical features of tick-borne relapsing fever in Tanzania. Consecutive patients with fever and spirochetes demonstrated in a thick blood smear at Haydom Lutheran Hospital from 1 January to 31 December 2003 underwent clinical and cerebrospinal fluid (CSF) examination. Forty-four patients were included, making an estimated minimum annual incidence of 11 per 100 000 population in this region. The mortality rate was 2.3% (95% CI = 0-12). The most frequent complaints were generalized malaise (93%), headache (86%), nausea and vomiting (52%). None of the patients [0% (95% CI = 0-8)] had focal neurological symptoms. Cell count in CSF was normal in 22 and slightly elevated (6-12 leukocytes/mm(3)) in 20 patients. Two of three pregnancies had a poor outcome. Jarisch-Herxheimer reactions, bleeding complications and ocular manifestations were infrequent. In conclusion, tick-borne relapsing fever is a common disease in the Northern highland of Tanzania, but in contrast to other parts of the world, neurological involvement is uncommon in this area. PMID- 15885050 TI - A randomized, double-blind, placebo-controlled study of the efficacy and safety of tolperisone in spasticity following cerebral stroke. AB - To study the efficacy and safety of tolperisone - a centrally acting muscle relaxant with membrane stabilizing activity - in the treatment of stroke-related spasticity. This was a randomized, double-blind, placebo-controlled, multicenter study with parallel groups. Treatment lasted 12 weeks and was started with a titration period of variable length (dose range 300-900 mg tolperisone daily). The degree of spasticity determined on the Ashworth Scale in the most severely affected joint area was defined as primary target parameter. Hundred and twenty patients (43 females, 77 males) in a mean age of 63.3 +/- 10.6 years were recruited and received treatment. In the majority of patients both limbs of each side (right: n = 59; left: n = 56) were affected by the spasticity which on average had been present for 3.3 +/- 4.4 years. A 62% of the patients were treated with a daily dose >/=600 mg tolperisone. Tolperisone reduced the mean Ashworth Score by a mean of 1.03 +/- 0.71 compared with a mean reduction of 0.47 +/- 0.54 in the placebo group (P < 0.0001). A 78.3% of the patients on tolperisone versus 45% of the placebo patients experienced a reduction by at least 1 point on the Ashworth Scale (P < 0.0001). Functional and overall assessments of efficacy confirmed superior efficacy of tolperisone. Adverse events occurred less often on active treatment (n = 19) than on placebo (n = 26) and were mostly of mild-to-moderate intensity. No withdrawals caused by adverse events were reported in the tolperisone group. The findings of the present study demonstrate the efficacy and excellent tolerance of tolperisone in the treatment of spastic hypertonia following cerebral stroke. Study data further suggest that an individual dose titration which may exceed the recommended maximum dose of 450 mg daily results in optimized therapeutic benefit. PMID- 15885051 TI - Doppler ultrasound and clinical findings in patients with acute ischemic stroke treated with intravenous thrombolysis. AB - The aim of this study was to assess cerebral hemodynamics in patients with acute ischemic stroke undergoing thrombolytic therapy and to assess the relationship between cerebral hemodynamics and outcome. Forty-one unselected patients admitted to hospital within 3 h received intravenous thrombolytic therapy and were examined by extracranial and transcranial Doppler ultrasound examinations. Their strokes were clinically graded with the National Institute of Health Stroke Scale. Outcome after 3 months was graded with the modified Rankin Scale. Amongst the 27 patients who had an additional ultrasound examination 24 h after treatment, favorable outcome was significantly more common amongst patients with recanalization than amongst those without (P < 0.004). Thirteen patients with middle cerebral artery occlusions were continuously monitored during thrombolysis and frequently up to 5 h after start of thrombolysis. Early recanalization occurred in nine (69%), at a median delay of 178 min (range 140-287) after stroke onset. All of these nine patients had a favorable outcome. Recanalization within 24 h was associated with favorable outcome. Subgroup analysis suggests that this effect is mostly related to early recanalization within the first 5 h after stroke. Transcranial Doppler may therefore help to identify those patients most probably to benefit from thrombolysis, especially in those patients with a higher potential risk of complications. PMID- 15885052 TI - Anterior and non-anterior ruptured aneurysms: memory and frontal lobe function performance following coiling. AB - Our aim was to compare memory and frontal function performance between two groups of patients treated with Guglielmi detachable coil (GDC) following intracranial ruptured aneurysm. The subgroups drawn following the localization of the aneurysm consisted of 19 patients presenting with anterior communicating artery aneurysms and 16 patients exhibiting middle cerebral artery and posterior communicating artery aneurysms. The 35 patients and 35 normal controls were administered extensive neuropsychological assessment. Additionally, a scale of qualitative changes of mood was presented to the patients and patients' relatives. The patients showed a better general performance on memory compared with the executive function performance, which was similarly impaired in both groups. We suggest that executive functions are more vulnerable to differently located lesions than memory functions. PMID- 15885053 TI - Extra temporal involvement in herpes simplex encephalitis. AB - Temporal lobe abnormalities on brain imaging have been described as strong evidence for herpes simplex encephalitis (HSE) in appropriate clinical settings. Extra temporal abnormalities are less well described in these patients. We retrospectively reviewed 20 patients of HSE and found extra temporal involvement in 11 (55%) patients. Three patients (15 %) had pure extra temporal abnormalities. Twelve patients (60%) had temporal lobe involvement, four patients (20%) had pure temporal lobe involvement and five patients (25%) had normal CT/MRI scans. Our study suggests that extra temporal involvement on brain imaging is common in HSE and in a significant minority of the patients this can even be the sole abnormality. PMID- 15885054 TI - Bone mineral density in familial amyloid polyneuropathy and in other neuromuscular disorders. AB - Neuromuscular diseases are a known risk factor for immobilization-induced osteoporosis. The aim of the study was to analyse bone mineral density (BMD) in patients with familial amyloid polyneuropathy (FAP) type I (Val30 Met) and to compare them with a population of patients with other neuromuscular disorders. We studied 24, ambulatory, neuromuscular patients, all men and premenopausal women. We included 12 FAP patients (GI) and 12 patients with other disorders (GII). Clinical data included age, sex, height, weight, alcohol intake, smoking, calcium intake, physical activity and history of fractures. Serum and urinary calcium, osteocalcin, bone alkaline phosphatase, parathyroid hormone, thyroid stimulating hormone and urinary N-telopeptide cross-linked type 1 collagen were determined in all patients. Bone mineral density of lumbar spine, hip and wrist were determined by dual energy X-ray absorptiometry scan. No statistical differences were found in clinical or analytic data between the two groups, except for body mass index and calciuria, which were lower in GI. In GI, 54.5% were osteoporotic, against 23.1% in GII (P = 0.04). Bone mineral density was lower in GI when compared with GII, and tended to decrease with disease duration. Decreased BMI and the early autonomic involvement in GI probably explain the results. The prevention and early treatment of osteoporosis, in FAP patients should be considered a priority. PMID- 15885055 TI - Analysis of antiepileptic drugs use at a university hospital in Croatia. AB - To analyse the consumption rate of new generation antiepileptic drugs (AEDs) compared with traditional AEDs at a university hospital in Croatia. Antiepileptic drugs use was analysed retrospectively for two consecutive years, 2001 and 2002 at Departments of Neurology, Paediatrics, Psychiatry and Neurosurgery at the University Hospital Centre (UHC) Rijeka. The results obtained are expressed as number of defined daily doses (DDDs) per 100 bed days, as proposed by the WHO. The use of new generation AEDs was represented by 2% in 2001 and 5% in 2002. Majority of AEDs administered was taken by the barbiturates in both years. A wide spectrum of AEDs has been used at the Department of Paediatrics. At the Department of Neurology and Psychiatry use of barbiturates and carbamazepine predominated. The use of new AEDs at UHC has increased during the investigation period but it is still rather low compared with traditional AEDs. The similarity of our results with the result of the leading Croatian university hospitals might represent general routine AED prescription in country. Nationally based guidelines may bring more appropriate and rational approach for usage of modern AED. This task should be supported and promoted by international and national neurology associations. PMID- 15885056 TI - Recurrent strokes in a young adult patient with Fabry's disease. PMID- 15885057 TI - First case report of Streptococcus suis septicaemia and meningitis from Greece. PMID- 15885062 TI - Epidural haemorrhage of the cervical spinal cord: a post-mortem artefact? AB - Spinal epidural haemorrhage is a rare entity that occurs uncommonly in adults and rarely in children. It has a typical clinical presentation, although to date, the cause for the majority of cases remains unknown. We present a series of cases where epidural haemorrhage was identified at post-mortem, principly to the cervical cord, in cases outside the age range usually reported for clinical epidural haemorrhage, and with no underlying pathology to account for the finding. We present a hypothesis for a post-mortem cause for this finding and consider that, in the absence of any other identifiable causation, then this is a post-mortem occurrence similar to that of the Prinsloo-Gordon artefact of the soft tissues of the neck. This finding must be interpreted with care so as not to make the mistaken diagnosis of a nonaccidental head injury based on its finding, especially in the absence of intracranial, cranial nerve, optic nerve or eye pathologies. PMID- 15885063 TI - The dorsal raphe nucleus in schizophrenia: a post mortem study of 5 hydroxytryptamine neurones. AB - The 5-hydroxytryptamine (5-HT, serotonin) system has been implicated in the pathophysiology and treatment of schizophrenia. In this study, we addressed the hypothesis that a deficit of 5-HT neurones, either inherited or acquired, is central to the developmental pathology of the disorder. We examined putative 5-HT neurones of the dorsal raphe nucleus (DRN) in post mortem, formalin-fixed tissue from 15 schizophrenic patients and 20 control subjects matched for age and gender. No significant difference was detected between these groups in the number or size (cross-sectional area or diameter) of tryptophan-hydroxylase immunoreactive cell profiles viewed in transverse sections collected from the level of the trochlear decussation to the emergence of the trigeminal nerve. Profile number was not affected by age, gender, side of the brainstem (left or right) or post mortem interval; however, time in formalin correlated negatively with the number of neurones counted. Moreover, a significant negative correlation was detected between time in formalin and the levels of immunoreaction product (optical density), which in turn correlated positively with our profile counts. A positive correlation was found between the age of subjects and our estimates of cell size. Our results do not support the proposal that an abnormality in the number and/or size of DRN 5-HT neurones is central to the aetiopathology of schizophrenia. PMID- 15885064 TI - The impact of argyrophilic grain disease on the development of dementia and its relationship to concurrent Alzheimer's disease-related pathology. AB - Argyrophilic grain disease (AGD) constitutes a neurodegenerative disorder that occurs in the brains of the elderly and affects 5% of all patients with dementia. Tau protein-containing lesions known as argyrophilic grains and located predominantly in limbic regions of the brain characterize this disease. Dementia is encountered in only a subset of cases that display the morphological pattern of AGD. The aim of this study is to determine the role of concurrent Alzheimer's disease (AD)-related pathology for the development of dementia in AGD patients. A total of 204 post-mortem brains from 30 demented and 49 nondemented AGD patients, 39 AD patients, and from 86 nondemented controls without AGD were staged for AD related neurofibrillary tangles (NFTs) as well as amyloid beta-protein (Abeta) deposition. To identify differences in AD-related pathology between demented and nondemented AGD cases, and to differentiate the pattern of AD-related changes in demented and nondemented AGD cases from that seen in AD and nondemented controls, we statistically compared the stages of Abeta and NFT distribution among these groups. Using a logistic regression model, we showed that AGD has a significant effect on the development of dementia beyond that attributable to AD-related pathology (P < 0.005). Demented AGD cases showed lower stages of AD-related pathology than did pure AD cases but higher stages than nondemented AGD patients. AGD associated dementia was seen in the presence of NFT (Braak)-stages II-IV and Abeta-phases 2-3, whereas those stages were not associated with dementia in the absence of AGD. In conclusion, AGD is a clinically relevant neurodegenerative entity that significantly contributes to the development of dementia by lowering the threshold for cognitive deficits in the presence of moderate amounts of AD related pathology. PMID- 15885065 TI - Distribution, characterization and clinical significance of microglia in glioneuronal tumours from patients with chronic intractable epilepsy. AB - Cells of the microglia/macrophage lineage represent an important component of different brain tumours. However, there is little information about the microglia/macrophage cell system in glioneuronal tumours and its possible contribution to the high epileptogenecity of these lesions. In the present study, the distribution of cells of the microglia/macrophage lineage was studied by immunocytochemistry for CD68 and human leucocyte antigen (HLA)-DR in a group of glioneuronal tumours, including gangliogliomas (GG, n = 30), and dysembryoplastic neuroepithelial tumours (DNT, n = 17), from patients with chronic intractable epilepsy. A significant number of microglia/macrophage cells were observed in the large majority of glioneuronal tumours, both within the tumour and in the peritumoral region. Activated microglial cells positive for HLA-DR were localized around blood vessels and clustered around tumour neuronal cells. The density of activated microglial cells correlated with the duration of epilepsy, as well as with the frequency of seizures prior to surgical resection. These observations indicate that the presence of cells of the microglial/macrophage cell system is a feature of glioneuronal tumours and is functionally related to epilepsy, either directly in epileptogenesis or through activation following seizure activity. PMID- 15885066 TI - Up-regulation of osteopontin and alphaBeta-crystallin in the normal-appearing white matter of multiple sclerosis: an immunohistochemical study utilizing tissue microarrays. AB - Tissue microarrays assembled from control and multiple sclerosis (MS) brain tissue have been used to assess the expression patterns and cellular distribution of two antigens, the proinflammatory cytokine osteopontin and the inducible heat shock protein alphaBeta-crystallin, which have previously been implicated in MS pathogenesis. Tissue cores were taken from paraffin-embedded donor blocks containing chronic active or chronic inactive plaques and normal-appearing white matter (NAWM) in seven MS cases, and white matter (WM) in five control cases. Expression patterns of both proteins were assessed against myelin density and microglial activation in the different tissue categories. Both proteins showed increased expression in all categories of MS tissue compared with control WM. The results indicate progressive up-regulation of expression of osteopontin with increased plaque activity, while elevation of alphaBeta-crystallin expression in MS tissue was independent of demyelination. In MS NAWM a significant correlation was observed between high levels of expression of osteopontin and alphaBeta crystallin. Osteopontin expression was predominantly confined to astrocytes throughout MS tissues. alphaBeta-crystallin was expressed on astrocytes, oligodendrocytes and occasionally on demyelinated axons. Taken together, these data indicate a wider distribution of osteopontin and alphaBeta-crystallin in MS tissues than previously described and support their proposed role in MS pathogenesis. PMID- 15885067 TI - The significance of beta-APP immunoreactivity in forensic practice. AB - The neuropathologist involved in forensic work is not uncommonly confronted with a case in which there is no or only a limited history or, if available, the information is uncertain or is often conflicting. In recent years the immunohistochemical stain beta-amyloid precursor protein (beta-APP) has been used to assess the extent of axonal injury in a variety of pathological processes but in forensic practice is of greatest utility in the assessment of traumatic brain injury. Diffuse traumatic axonal injury (TAI) in humans has been demonstrated by beta-APP immunoreactivity in patients surviving at least 2 h after head injury. However, many of these patients also have an associated ischaemic injury, either focal or diffuse, which may make the interpretation of beta-APP immunoreactivity difficult. The present study was designed to evaluate if the published descriptions of the different morphological patterns and distributions of beta APP immunoreactive axons could be used to microscopically distinguish axonal injury attributed to trauma from other causes. To test this hypothesis a total of 73 cases were reviewed. The cases were selected from six different groups based on clinical information. Immunostained sections from each case were assessed 'blind' to the clinical history, and the microscopic pattern and distribution of beta-APP positive axons were recorded. Haematoxylin and eosin (H+E) stained sections were then reviewed for each case and a final pathological diagnosis was recorded and compared to the clinical history. 62/73 (85%) cases were correctly correlated with the clinical history and in particular 14/17 (82%) cases of TAI were correctly identified. These findings indicate that the published microscopic patterns of the distribution of beta-APP positive axons in TAI and in diffuse ischaemic injury can be used, in conjunction with microscopy of H+E stained sections to determine the cause of axonal pathology in most cases. PMID- 15885068 TI - The Alzheimer disease-related calcium-binding protein Calmyrin is present in human forebrain with an altered distribution in Alzheimer's as compared to normal ageing brains. AB - The EF-hand calcium binding protein Calmyrin (also called CIB-1) was shown to interact with presenilin-2 (PS-2), suggesting that this interaction might play a role in the pathogenesis of Alzheimer's disease (AD). Here we have investigated the distribution of Calmyrin in normal human and AD brain. In normal brain Calmyrin immunoreactivity was unevenly distributed with immunostaining in pyramidal neurones and interneurones of the palaeo-cortex and neocortex, cerebellar granule cells and hypothalamic neurones of the paraventricular, ventromedial and arcuate nuclei. Moderate immunoreactivity was present in hippocampal pyramidal cells and stronger in dentate gyrus neurones. Thalamic and septal neurones were devoid of immunoreactivity. No apparent differences were visible between stainings of brain sections from younger and older nondemented patients. In AD brain a substantial loss of Calmyrin-immunopositive neurones was observed in all regions, especially in cortical areas. Still immunoreactive neurones, however, displayed stronger staining that was especially concentrated in perinuclear regions. Calmyrin immunosignals were in part associated with diffuse and senile plaques. Thus, although protein levels of Calmyrin are low in human forebrain, its cellular localization as well as its altered distribution in AD brain suggest that it may be involved in the pathogenesis of AD. PMID- 15885069 TI - Does drug abuse alter microglial phenotype and cell turnover in the context of advancing HIV infection? AB - The aim of this study was to test the effects of drug abuse, in particular opiate abuse, on the phenotype and turnover of microglial cells within the brain in the context of advancing HIV infection. Basal ganglia and hippocampus sections were studied in 51 cases divided into six groups: HIV-negative normal controls, HIV negative drug abusers, AIDS nondrug abusers, AIDS drug abusers, HIV encephalitis (HIVE) nondrug abusers and HIVE drug abusers. None of the cases studied had received highly active anti-retroviral therapy (HAART). Microglial phenotypes were defined using CD14, CD16, CD68 and major histocompatibility class II (MHC II). Microglial turnover was assessed using terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) (DNA damage), BAX (proapoptotic marker), Fas (CD95) (proapoptotic), proliferating cell nuclear antigen (PCNA) (proliferation and DNA repair), Ki-67 (cell proliferation) and BCL-2 (antiapoptosis). We find increased expression of MHC II and CD16 in response to drug abuse. We also noted increased levels of TUNEL positivity in drug abusers compared to nondrug abusers, although conversely we found lower levels of BAX in those who had abused drugs. We find no evidence of microglial proliferation in any of our study groups, including HIVE, although HIV infection leads to increased expression of CD16, CD68 and MHC II. CD14 expression was restricted to perivascular microglia in all groups (including normal controls) apart from the two HIVE groups where some but not all cases also showed parenchymal expression of CD14. In contrast, CD16 was found in parenchymal microglia in all groups. Using high-pressure antigen retrieval and tyramide signal amplification, we find moderately high levels of CD16 expression in the parenchyma of normal brains which is not normally observed using standard avidin/biotin complex (ABC) techniques. This suggests that a low basal expression of CD16 occurs in many resident microglial cells which may potentially be upregulated in HIV-infected individuals. From these data, we suggest that not all the CD16+ parenchymal cells detected in AIDS brains (using ABC) represent influx of monocyte lineage cells from the blood. Finally the increased expression of MHC II and CD68 detected in drug abusers with HIVE compared to nondrug abusers with HIVE suggests that the combination of drug abuse and HIV infection has a greater deleterious effect on the brain than either individual insult on its own. PMID- 15885070 TI - The basement membrane at the tumour-brain interface of brain-invasive grade I meningiomas. PMID- 15885073 TI - Abstracts of the XV International Symposium on Drugs Affecting Lipid Metabolism, 24-27 October, 2004, Venice, Italy. PMID- 15885075 TI - E6* oncoprotein expression of human papillomavirus type-16 determines different ultraviolet sensitivity related to glutathione and glutathione peroxidase antioxidant defence. AB - Clinical observations of non-melanoma skin cancer in immunocompromised patients, such as organ transplant recipients, suggest co-operative effects of human papillomavirus (HPV) and ultraviolet (UV) radiation. The aim of the present study is to evaluate UV sensitivity and DNA damage formation according to antioxidant status in HPV16-infected keratinocytes. We used SKv cell lines, infected with HPV16 and well characterized for their proliferative and tumorigenic capacities. We showed that SKv cell lines presented various E6* (a truncated form of E6) RNA levels. We demonstrated that the higher oncoprotein RNA expression level was associated with a higher resistance to solar-simulated radiation, more specifically to UVB radiation and to hydrogen peroxide. Moreover, this high resistance was associated with a low oxidative DNA damage formation after UV radiation and was related to high glutathione content and glutathione peroxidase activities. Therefore, the results of our study suggest that E6* levels could modulate the glutathione/glutathione peroxidase pathway providing a mechanism to protect HPV-infected keratinocytes against an environmental oxidative stress, such as UV radiation. PMID- 15885076 TI - Selective down-regulation of the alpha6-integrin subunit in melanocytes by UVB light. AB - In vivo, melanocytes bind to laminin (LM) molecules of the basement membrane (BM) via the integrins alpha3beta1 and alpha6beta1, and they adhere to neighbouring keratinocytes via E-cadherin. Only few studies have addressed the impact of ultraviolet (UV) light on the interaction of melanocytes with their microenvironment. In this report, we examined the influence of UVB irradiation on the expression of the most important melanocyte-adhesion molecules (E-, N cadherin, alpha2-, alpha3-, alpha5-, alpha6-, alphaV-, beta1-, beta3-integrins and ICAM-1) in vitro by flow cytometry. We were able to demonstrate that the alpha6-integrin subunit is selectively and reversibly down-regulated by UVB in a dwzm 150ose-dependent manner. In comparison, keratinocytes lacked UVB-inducible alterations in the expression of alpha6-integrin. In the presence of LM-1, the UVB-induced down-regulation of alpha6-integrin in melanocytes was significantly reduced. Moreover, LM-1 increased the resistance of melanocytes to UVB-induced cell death, as measured by annexinV-binding analysis. This effect was reversed by preincubation with an alpha6-integrin-blocking antibody. By immunofluorescence, we could demonstrate that UVB leads to a dose-dependent internalization of alpha6 integrin, providing an obvious explanation for the down-regulation on the outer cell surface observed by flow cytometry. We suggest that adhesion to LM-1 through alpha6-integrin represents a protective mechanism for melanocytes to withstand UVB damage. Through alpha6-integrin internalization, sunburns might alter the interaction between melanocytes and the BM, resulting in apoptosis induced by loss of anchorage (anoikis). Repeated sunburns may then lead to the selection of a population of melanocytes which are capable of anchorage-independent survival, culminating in solar nevogenesis and melanoma development. PMID- 15885077 TI - Expression of hurpin, a serine proteinase inhibitor, in normal and pathological skin: overexpression and redistribution in psoriasis and cutaneous carcinomas. AB - Hurpin was identified by differential display analysis studying UV-repressible genes in the keratinocyte cell line HaCaT. We have previously reported that hurpin mRNA is overexpressed in psoriatic skin compared to non-lesional or normal skin; hurpin inhibits cathepsin L and that, after overexpression in keratinocytes, hurpin decreases UV-induced apoptosis. To further study the expression of hurpin, we have isolated monoclonal antibodies against hurpin and analyzed its expression in normal and diseased skin by immunohistochemistry (IHC). In the epidermis of normal skin, we found hurpin to be mainly expressed in the stratum basale. In contrast, we found an enhanced expression of hurpin in the stratum spinosum and stratum granulosum in the majority of diseased skin samples. Within the dermis of normal and diseased skin, hurpin was detected in sebaceous and sweat glands, hair follicles, and endothelial cells of blood vessels. Hurpin was localized in the cytoplasm in normal and diseased skin. Additionally to IHC, we analyzed hurpin expression in selected skin diseases by semiquantitative reverse-transcription polymerase chain reaction. We found overexpression of hurpin mRNA in psoriasis, squamous cell carcinoma (SCC), and actinic keratosis. In contrast, expression of hurpin in melanoma and basal cell carcinoma was comparable to that in normal skin. Overall, the strongest overexpression was observed in SCC and psoriasis. Individual differences in hurpin expression between patients were observed. The increased expression and redistribution of hurpin in diseased skin suggests its possible involvement in inflammatory processes or the regulation of endogenous or pathogen-derived proteinase activity. Additional studies will elucidate the physiological role of hurpin. PMID- 15885078 TI - 5-aminolaevulinic acid photodynamic therapy in a transgenic mouse model of skin melanoma. AB - Photodynamic therapy (PDT) is widely used to treat preneoplastic skin lesions and non-melanoma skin tumours. Studies analyzing the effects of PDT on malignant melanoma have yielded conflicting results. On the one hand, melanoma cell lines in culture as well as cell lines transplanted into experimental animals were sensitive to PDT. On the other hand, spontaneous melanomas of human patients responded poorly to most PDT regimens tested so far. Here, we analyzed effects of 5-aminolaevulinic acid (5-ALA)-based PDT on melanoma cell lines and on experimental melanomas. To mimic the clinical situation as closely as possible, metallothionein-I/ret (MT-ret) mice, a transgenic model of skin melanoma development, were used. Optimal doses of 5-ALA as well as energy doses and power densities were determined in vitro using a cell line (Mel25) established by us from a melanoma of an MT-ret transgenic mouse as well as commercially available human and mouse melanoma cell lines. Treatment with light irradiation alone had no effect. In combination with 5-ALA, however, this illumination readily induced the death of all mouse and human melanoma cell lines examined. Still, 5-ALA PDT caused only minor focal regressive changes including haemorrhages and fibrosis of MT-ret melanomas in vivo and did not significantly delay tumour growth. These results show that, even though MT-ret melanoma cells are vulnerable to 5-ALA PDT in vitro, malignant MT-ret melanomas in vivo are quite resistant to this type of therapy at doses which are highly effective in vitro. PMID- 15885079 TI - Dermal mast cells play a central role in the incidence of scratching behavior in mice induced by multiple application of the hapten, 2,4,6-trinitrochlorobenzene. AB - Repeated application of 1% 2,4,6-trinitrochlorobenzene (TNCB) in acetone solution causes chronic skin inflammation in BALB/c mice. Associated scratching behavior gradually appeared, and chronic scratching behavior was established over 40 days after the initial application of TNCB. In order to explore the possible involvement of T cells and mast cells in the appearance of pruritus, we examined the response of athymic nude mice and genetically mast-cell-deficient mice. We could not detect either severe skin inflammation or immunoglobulin (Ig)E production in T-cell-deficient BALB/c nu/nu mice even after 80 days of TNCB treatment, whereas typical severe skin inflammation and IgE production were observed in mast-cell-deficient WBB6F1-W/Wv and WBB6F1-Sl/Sld mice. Furthermore, we observed persistent scratching behavior in WBB6F1-W/Wv mice, but not in BALB/c nu/nu and WBB6F1-Sl/Sld mice. Histological examination of TNCB-treated animals revealed the development of dermal mast cells in W/Wv mice but not in Sl/Sld mice. Degranulation of dermal mast cells was observed in the WBB6F1-W/Wv genotype, but most mast cells remained intact in TNCB-treated BALB/c nu/nu mice. These results suggest that mast cells play a pivotal role in the incidence of scratching behavior in this chronic pruritus model. PMID- 15885080 TI - Non-lesional vitiliginous melanocytes are not characterized by an increased proneness to nitric oxide-induced apoptosis. AB - Nitric oxide (NO) is a reactive endogenous molecule with multiple functions including inflammation and immunity. NO stimulates melanogenesis by activating soluble guanylyl cyclase (sGC) resulting in increases in intracellular guanosine 3',5'-cyclic monophosphate (cGMP). In vitro experiments showed that NO could inhibit the de novo attachment of melanocytes to extracellular matrix (ECM) suggesting that NO-induced aberrant perturbation of melanocyte-ECM interaction could be a reason for melanocyte loss in vitiliginous lesions. Here, we examined whether there might be differences between normal melanocytes and vitiliginous melanocytes (VMs) with respect to NO-induced detachment from ECM and whether cGMP is involved. We used the direct NO donor (Z)-1-[N-(3-ammoniopropyl)-N-(n propyl)amino]diazen-1-ium-1,2-diolate and the peroxynitrite donor 3-morpholino sydnonimine for the present studies. These donors induced detachment of both normal melanocytes and non-lesional VMs in a time- and concentration-dependent manner with comparable susceptibility and similar expression profile of sGC. Treatment of melanocytes with caspase inhibitors reduced cell detachment, indicating that a major part of the detachment is due to apoptosis. The NO induced detachment but not apoptosis was partly inhibited in the presence of sGC and cGMP-dependent protein kinase inhibitors. In addition, the membrane-permeable cGMP analog 8-(4-chlorophenyethio/guanosine-3',5'-cyclic monophosphate (PCPT) cGMP was not able to induce apoptosis in melanocytes, suggesting that NO-induced detachment of melanocytes via apoptosis is cGMP-independent. The present results also indicate that there are no apparent differences between NO-induced detachment of non-lesional vitiliginous and normal melanocytes from ECM. PMID- 15885081 TI - The expression of human leukocyte antigen-DR and CD25 on circulating T cells in cutaneous lupus erythematosus and correlation with disease activity. AB - BACKGROUND: Lupus erythematosus (LE) is a chronic autoimmune disease with a broad clinical spectrum reaching from primarily cutaneous manifestations [cutaneous LE (CLE)] up to systemic disease [systemic LE (SLE)]. In patients with SLE, the expression of activation markers on circulating T cells reflects disease activity. Here, we investigated whether this also holds true for patients with CLE. PATIENTS AND METHODS: The expression of the activation markers human leukocyte antigen (HLA)-DR and CD25 on circulating T lymphocytes was measured by flow cytometry in 24 patients suffering from different types of active CLE. Simultaneously, the disease activity was assessed clinically using a CLE activity index. Eighteen healthy donors were analyzed for control purposes. RESULTS: HLA DR was expressed on a significantly elevated percentage of both CD4+ and CD8+ circulating T cells in active CLE patients when compared with healthy controls. The percentage of HLA-DR-expressing T lymphocytes closely correlated with the disease activity. Interestingly, in disseminated scarring chronic discoid LE, a significantly increased percentage of CD25+ cells was observed only in the subset of skin-homing cutaneous lymphocyte antigen (CLA)+CD4+ and CD8+ T cells. CONCLUSION: Our results provide evidence that activation markers on peripheral blood T cells might help to objectively assess the disease activity in CLE. Furthermore, a significant population of CD25+CLA+CD8+ T cells can only be detected in a subgroup of patients with disseminated scarring CLE and might reflect the systemic expansion of activated cytotoxic T lymphocytes involved in destruction of epidermal tissue. PMID- 15885082 TI - Atopic dermatitis-like pruritic skin inflammation caused by feeding a special diet to HR-1 hairless mice. AB - Dry skin/barrier dysfunction is considered to be one of the characteristic features of atopic dermatitis (AD). When HR-1 hairless mice are fed a special diet, HR-AD, dry red skin is induced. We examined whether HR-AD-fed mouse could be used as a model for AD by showing itch-associated scratching behaviour and by analysing the immunological change. HR-1 mice were fed HR-AD from 4 weeks old. HR AD-fed mice showed severe dry skin symptoms accompanied by a decrease in dermal water content and an increase in transepidermal water loss and prolonged scratching bout duration on day 14 or 28. These symptoms became gradually worse until day 56. Marked epidermal hyperplasia and slight increase in CD4+ cells in the skin were observed from day 28. In contrast, increases in circulating T cells and serum immunoglobulin E were seen from day 41. Other skin-infiltrating inflammatory cells, such as eosinophils and mast cells, were increased on day 56 but not on day 28. Though daily oral treatment with dexamethasone reduced the increased numbers of these cells, it did not affect the dry skin symptoms or the prolonged scratching episodes. In contrast, the development of dry skin was inhibited by feeding with 10% normal diet-containing HR-AD. The skin barrier dysfunction in HR-AD-fed mice is closely associated with the development of AD like pruritus. Changes in the immunological parameters observed may be the consequence of skin barrier dysfunction. Our findings suggest that HR-AD-fed mouse could be used as a dry skin-based experimental model for AD. PMID- 15885083 TI - Production of large numbers of Langerhans' cells with intraepithelial migration ability in vitro. AB - Langerhans' cells (LCs) are a subset of immature dendritic cells (DCs) and play a key role in the initiation and regulation of immune responses. Functional studies of these cells have been hampered by difficulties in generating a large number of LCs in vitro. We describe a new method to efficiently generate immature DCs exhibiting morphological, immunohistochemical, and ultrastructural features of LCs (CD1a+, Birbeck Granules+, CD207+, E-cadherin+, cutaneous lymphocyte associated antigen+, and CCR6+) from a limited number of CD34+ cord blood progenitors. This method is based on a two-step procedure consisting of an amplification phase followed by a terminal differentiation induction. The amplification step is initiated with a combination of hematopoietic growth factors (thrombopoietin/stem cell factor/fetal liver tyrosine kinase-3 ligand), cytokines (granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and interleukin-4), and 5 ng/ml of transforming growth factor (TGF) beta1. The differentiation is induced by increasing the concentration of TGF beta1 to 12.5 ng/ml. These culture conditions were efficient for generating a large number of immature LCs (8.74 x 10(6) +/- 3.2) from 15 x 10(4) CD34+ progenitor cells. In addition, these LCs were shown to be able to infiltrate an in vitro reconstructed epithelium. Because LCs play an important role in the mucosal immunity, this technique could be useful to study their interactions with epithelial pathogenic agents and to perform pharmacological, toxicological, and clinical research. PMID- 15885084 TI - How to treat that nasty itch. PMID- 15885086 TI - Nature, nurture and neurology: gene-environment interactions in neurodegenerative disease. FEBS Anniversary Prize Lecture delivered on 27 June 2004 at the 29th FEBS Congress in Warsaw. AB - Neurodegenerative disorders, such as Huntington's, Alzheimer's, and Parkinson's diseases, affect millions of people worldwide and currently there are few effective treatments and no cures for these diseases. Transgenic mice expressing human transgenes for huntingtin, amyloid precursor protein, and other genes associated with familial forms of neurodegenerative disease in humans provide remarkable tools for studying neurodegeneration because they mimic many of the pathological and behavioural features of the human conditions. One of the recurring themes revealed by these various transgenic models is that different diseases may share similar molecular and cellular mechanisms of pathogenesis. Cellular mechanisms known to be disrupted at early stages in multiple neurodegenerative disorders include gene expression, protein interactions (manifesting as pathological protein aggregation and disrupted signaling), synaptic function and plasticity. Recent work in mouse models of Huntington's disease has shown that enriching the environment of transgenic animals delays the onset and slows the progression of Huntington's disease-associated motor and cognitive symptoms. Environmental enrichment is known to induce various molecular and cellular changes in specific brain regions of wild-type animals, including altered gene expression profiles, enhanced neurogenesis and synaptic plasticity. The promising effects of environmental stimulation, demonstrated recently in models of neurodegenerative disease, suggest that therapy based on the principles of environmental enrichment might benefit disease sufferers and provide insight into possible mechanisms of neurodegeneration and subsequent identification of novel therapeutic targets. Here, we review the studies of environmental enrichment relevant to some major neurodegenerative diseases and discuss their research and clinical implications. PMID- 15885087 TI - Biogenesis of peroxisomes. Topogenesis of the peroxisomal membrane and matrix proteins. AB - Genetic and proteomic approaches have led to the identification of 32 proteins, collectively called peroxins, which are required for the biogenesis of peroxisomes. Some are responsible for the division and inheritance of peroxisomes; however, most peroxins have been implicated in the topogenesis of peroxisomal proteins. Peroxisomal membrane and matrix proteins are synthesized on free ribosomes in the cytosol and are imported post-translationally into pre existing organelles (Lazarow PB & Fujiki Y (1985) Annu Rev Cell Biol1, 489-530). Progress has been made in the elucidation of how these proteins are targeted to the organelle. In addition, the understanding of the composition of the peroxisomal import apparatus and the order of events taking place during the cascade of peroxisomal protein import has increased significantly. However, our knowledge on the basic principles of peroxisomal membrane protein insertion or translocation of peroxisomal matrix proteins across the peroxisomal membrane is rather limited. The latter is of particular interest as the peroxisomal import machinery accommodates folded, even oligomeric, proteins, which distinguishes this apparatus from the well characterized translocons of other organelles. Furthermore, the origin of the peroxisomal membrane is still enigmatic. Recent observations suggest the existence of two classes of peroxisomal membrane proteins. Newly synthesized class I proteins are directly targeted to and inserted into the peroxisomal membrane, while class II proteins reach their final destination via the endoplasmic reticulum or a subcompartment thereof, which would be in accord with the idea that the peroxisomal membrane might be derived from the endoplasmic reticulum. PMID- 15885088 TI - Chain initiation on type I modular polyketide synthases revealed by limited proteolysis and ion-trap mass spectrometry. AB - Limited proteolysis in combination with liquid chromatography-ion trap mass spectrometry (LC-MS) was used to analyze engineered or natural proteins derived from a type I modular polyketide synthase (PKS), the 6-deoxyerythronolide B synthase (DEBS), and comprising either the first two extension modules linked to the chain-terminating thioesterase (TE) (DEBS1-TE); or the last two extension modules (DEBS3) or the first extension module linked to TE (diketide synthase, DKS). Functional domains were released by controlled proteolysis, and the exact boundaries of released domains were obtained through mass spectrometry and N terminal sequencing analysis. The acyltransferase-acyl carrier protein required for chain initiation (AT(L)-ACP(L)), was released as a didomain from both DEBS1 TE and DKS, as well as the off-loading TE as a didomain with the adjacent ACP. Mass spectrometry was used successfully to monitor in detail both the release of individual domains, and the patterns of acylation of both intact and digested DKS when either propionyl-CoA or n-butyryl-CoA were used as initiation substrates. In particular, both loading domains and the ketosynthase domain of the first extension module (KS1) were directly observed to be simultaneously primed. The widely available and simple MS methodology used here offers a convenient approach to the proteolytic mapping of PKS multienzymes and to the direct monitoring of enzyme-bound intermediates. PMID- 15885089 TI - Quantitative assessment of the glyoxalase pathway in Leishmania infantum as a therapeutic target by modelling and computer simulation. AB - The glyoxalase pathway of Leishmania infantum was kinetically characterized as a trypanothione-dependent system. Using time course analysis based on parameter fitting with a genetic algorithm, kinetic parameters were estimated for both enzymes, with trypanothione derived substrates. A K(m) of 0.253 mm and a V of 0.21 micromol.min(-1).mg(-1)for glyoxalase I, and a K(m) of 0.098 mm and a V of 0.18 micromol.min(-1).mg(-1) for glyoxalase II, were obtained. Modelling and computer simulation were used for evaluating the relevance of the glyoxalase pathway as a potential therapeutic target by revealing the importance of critical parameters of this pathway in Leishmania infantum. A sensitivity analysis of the pathway was performed using experimentally validated kinetic models and experimentally determined metabolite concentrations and kinetic parameters. The measurement of metabolites in L. infantum involved the identification and quantification of methylglyoxal and intracellular thiols. Methylglyoxal formation in L. infantum is nonenzymatic. The sensitivity analysis revealed that the most critical parameters for controlling the intracellular concentration of methylglyoxal are its formation rate and the concentration of trypanothione. Glyoxalase I and II activities play only a minor role in maintaining a low intracellular methylglyoxal concentration. The importance of the glyoxalase pathway as a therapeutic target is very small, compared to the much greater effects caused by decreasing trypanothione concentration or increasing methylglyoxal concentration. PMID- 15885090 TI - The lipid/protein interface as xenobiotic target site. Kinetic analysis of tadpole narcosis. AB - High-resolution X-ray diffraction structures of integral membrane proteins have revealed various binding modes of lipids, but current spectroscopic studies still use uniform macroscopic binding constants to describe lipid binding. The Adair approach employing microscopic lipid-binding constants has previously been taken to explain the enhancement of agonist binding to the nicotinic acetylcholine receptor by general anaesthetics in terms of the competitive displacement of essential lipid activator molecules [Walcher S, Altschuh J & Sandermann H (2001) J. Biol. Chem.276, 42191-42195]. This approach was extended to tadpole narcosis induced by alcohols. A single class, or two different classes of lipid activator binding sites, are considered. Microscopic lipid and inhibitor binding constants are derived and allow a close fit to dose-response curves of tadpole narcosis on the basis of a preferential displacement of more loosely bound essential lipid activator molecules. This study illustrates the potential of the Adair approach to resolve protein-bound lipid populations. PMID- 15885091 TI - Production and utilization of hydrogen peroxide associated with melanogenesis and tyrosinase-mediated oxidations of DOPA and dopamine. AB - The synthesis and involvement of H(2)O(2) during the early stages of melanogenesis involving the oxidations of DOPA and dopamine (diphenolase activity) were established by two sensitive and specific electrochemical detection systems. Catalase-treated reaction mixtures showed diminished rates of H(2)O(2) production during the autoxidation and tyrosinase-mediated oxidation of both diphenols. Inhibition studies with the radical scavenger resveratrol revealed the involvement in these reactions of additional reactive intermediate of oxygen (ROI), one of which appears to be superoxide anion. There was no evidence to suggest that H(2)O(2) or any other ROI was produced during the tyrosinase-mediated conversion of tyrosine to DOPA (monophenolase activity). Establishing by electrochemical methods the endogenous production H(2)O(2) in real time confirms recent reports, based in large part on the use of exogenous H(2)O(2), that tyrosinase can manifest both catalase and peroxidase activities. The detection of ROI in tyrosinase-mediated in vitro reactions provides evidence for sequential univalent reductions of O(2), most likely occurring at the enzyme active site copper. Collectively, these observations focus attention on the possible involvement of peroxidase-H(2)O(2) systems and related ROI-mediated reactions in promoting melanocytotoxic and melanoprotective processes. PMID- 15885092 TI - Oligosaccharide synthesis in Fibrobacter succinogenes S85 and its modulation by the substrate. AB - In this article we compared the metabolism of phosphorylated and unphosphorylated oligosaccharides (cellodextrins and maltodextrins) in Fibrobacter succinogenes S85 resting cells incubated with the following substrates: glucose; cellobiose; a mixture of glucose and cellobiose; and cellulose. Intracellular and extracellular media were analysed by (1)H-NMR and by TLC. The first important finding is that no cellodextrins were found to accumulate in the extracellular media of cells, regardless of the substrate; this contrasts to what is generally reported in the literature. The second finding of this work is that maltodextrins of degree of polymerization > 2 are synthesized regardless of the substrate, and can be used by the bacteria. Maltotriose plays a key role in this metabolism of maltodextrin. Maltodextrin-1-phosphate was detected in all the incubations, and a new metabolite, corresponding to a phosphorylated glucose derivative, was produced in the extracellular medium when cells were incubated with cellulose. The accumulation of these phosphorylated sugars increased with the degree of polymerization of the substrate. PMID- 15885093 TI - Characterization of a prokaryotic haemerythrin from the methanotrophic bacterium Methylococcus capsulatus (Bath). AB - For a long time, the haemerythrin family of proteins was considered to be restricted to only a few phyla of marine invertebrates. When analysing differential protein expression in the methane-oxidizing bacterium, Methylococcus capsulatus (Bath), grown at a high and low copper-to-biomass ratio, respectively, we identified a putative prokaryotic haemerythrin expressed in high-copper cultures. Haemerythrins are recognized by a conserved sequence motif that provides five histidines and two carboxylate ligands which coordinate two iron atoms. The diiron site is located in a hydrophobic pocket and is capable of binding O(2). We cloned the M. capsulatus haemerythrin gene and expressed it in Escherichia coli as a fusion protein with NusA. The haemerythrin protein was purified to homogeneity cleaved from its fusion partner. Recombinant M. capsulatus haemerythrin (McHr) was found to fold into a stable protein. Sequence similarity analysis identified all the candidate residues involved in the binding of diiron (His22, His58, Glu62, His77, His81, His117, Asp122) and the amino acids forming the hydrophobic pocket in which O(2) may bind (Ile25, Phe59, Trp113, Leu114, Ile118). We were also able to model a three-dimensional structure of McHr maintaining the correct positioning of these residues. Furthermore, UV/vis spectrophotometric analysis demonstrated the presence of conjugated diiron atoms in McHr. A comprehensive genomic database search revealed 21 different prokaryotes containing the haemerythrin signature (PROSITE 00550), indicating that these putative haemerythrins may be a conserved prokaryotic subfamily. PMID- 15885094 TI - The effect of replacing the axial methionine ligand with a lysine residue in cytochrome c-550 from Paracoccus versutus assessed by X-ray crystallography and unfolding. AB - The structure of cytochrome c-550 from the nonphotosynthetic bacteria Paraccocus versutus has been solved by X-ray crystallography to 1.90 A resolution, and reveals a high structural homology to other bacterial cytochromes c(2). The effect of replacing the axial heme-iron methionine ligand with a lysine residue on protein structure and unfolding has been assessed using the M100K variant. From X-ray structures at 1.95 and 1.55 A resolution it became clear that the amino group of the lysine side chain coordinates to the heme-iron. Structural differences compared to the wild-type protein are confined to the lysine ligand loop connecting helices four and five. In the heme cavity an additional water molecule is found which participates in an H-bonding interaction with the lysine ligand. Under cryo-conditions extra electron density in the lysine ligand loop is revealed, leading to residues K97 to T101 being modeled with a double main-chain conformation. Upon unfolding, dissociation of the lysine ligand from the heme iron is shown to be pH dependent, with NMR data consistent with the occurrence of a ligand exchange mechanism similar to that seen for the wild-type protein. PMID- 15885095 TI - Interaction of synthetic peptides corresponding to hepatitis G virus (HGV/GBV-C) E2 structural protein with phospholipid vesicles. AB - The interaction with phospholipid bilayers of two synthetic peptides with sequences corresponding to a segment next to the native N-terminus and an internal region of the E2 structural hepatitis G virus (HGV/GBV-C) protein [E2(7 26) and E2(279-298), respectively] has been characterized. Both peptides are water soluble but associate spontaneously with bilayers, showing higher affinity for anionic than zwitterionic membranes. However, whereas the E2(7-26) peptide is hardly transferred at all from water to the membrane interface, the E2(279-298) peptide is able to penetrate into negatively charged bilayers remaining close to the lipid/water interface. The nonpolar environment clearly induces a structural transition in the E2(279-298) peptide from random coil to alpha-helix, which causes bilayer perturbations leading to vesicle permeabilization. The results indicate that this internal segment peptide sequence is involved in the fusion of HGV/GBV-C to membrane. PMID- 15885096 TI - Solution structure and backbone dynamics of the XPC-binding domain of the human DNA repair protein hHR23B. AB - Human cells contain two homologs of the yeast RAD23 protein, hHR23A and hHR23B, which participate in the DNA repair process. hHR23B houses a domain (residues 277 332, called XPCB) that binds specifically and directly to the xeroderma pigmentosum group C protein (XPC) to initiate nucleotide excision repair (NER). This domain shares sequence homology with a heat shock chaperonin-binding motif that is also found in the stress-inducible yeast phosphoprotein STI1. We determined the solution structure of a protein fragment containing amino acids 275-342 of hHR23B (termed XPCB-hHR23B) and compared it with the previously reported solution structures of the corresponding domain of hHR23A. The periodic positioning of proline residues in XPCB-hHR23B produced kinked alpha helices and assisted in the formation of a compact domain. Although the overall structure of the XPCB domain was similar in both XPCB-hHR23B and XPCB-hHR23A, the N-terminal part (residues 275-283) of XPCB-hHR23B was more flexible than the corresponding part of hHR23A. We tried to infer the characteristics of this flexibility through (15)N-relaxation studies. The hydrophobic surface of XPCB-hHR23B, which results from the diverse distribution of N-terminal region, might give rise to the functional pleiotropy observed in vivo for hHR23B, but not for hHR23A. PMID- 15885097 TI - Characterization of structural and catalytic differences in rat intestinal alkaline phosphatase isozymes. AB - To understand the differences between the rat intestinal alkaline phosphatase isozymes rIAP-I and rIAP-II, we constructed structural models based on the previously determined crystal structure for human placental alkaline phosphatase (hPLAP). Our models of rIAP-I and rIAP-II displayed a typical alpha/beta topology, but the crown domain of rIAP-I contained an additional beta-sheet, while the embracing arm region of rIAP-II lacked the alpha-helix, when each model was compared to hPLAP. The representations of surface potential in the rIAPs were predominantly positive at the base of the active site. The coordinated metal at the active site was predicted to be a zinc triad in rIAP-I, whereas the typical combination of two zinc atoms and one magnesium atom was proposed for rIAP-II. Using metal-depleted extracts from rat duodenum or jejunum and hPLAP, we performed enzyme assays under restricted metal conditions. With the duodenal and jejunal extract, but not with hPLAP, enzyme activity was restored by the addition of zinc, whereas in nonchelated extracts, the addition of zinc inhibited duodenal IAP and hPLAP, but not jejunal IAP. Western blotting revealed that nearly all of the rIAP in the jejunum extracts was rIAP-I, whereas in duodenum the percentage of rIAP-I (55%) correlated with the degree of AP activation (60% relative to that seen with jejunal extracts). These data are consistent with the presence of a triad of zinc atoms at the active site of rIAP-I, but not rIAP-II or hPLAP. Although no differences in amino acid alignment in the vicinity of metal-binding site 3 were predicted between the rIAPs and hPLAP, the His153 residue of both rIAPs was closer to the metal position than that in hPLAP. Between the rIAPs, a difference was observed at amino acid position 317 that is indirectly related to the coordination of the metal at metal-binding site 3 and water molecules. These findings suggest that the side-chain position of His153, and the alignment of Q317, might be the major determinants for activation of the zinc triad in rIAP-I. PMID- 15885098 TI - Adaptation of intronic homing endonuclease for successful horizontal transmission. AB - Group I introns are thought to be self-propagating mobile elements, and are distributed over a wide range of organisms through horizontal transmission. Intron invasion is initiated through cleavage of a target DNA by a homing endonuclease encoded in an open reading frame (ORF) found within the intron. The intron is likely of no benefit to the host cell and is not maintained over time, leading to the accumulation of mutations after intron invasion. Therefore, regular invasional transmission of the intron to a new species at least once before its degeneration is likely essential for its evolutionary long-term existence. In many cases, the target is in a protein-coding region which is well conserved among organisms, but contains ambiguity at the third nucleotide position of the codon. Consequently, the homing endonuclease might be adapted to overcome sequence polymorphisms at the target site. To address whether codon degeneracy affects horizontal transmission, we investigated the recognition properties of a homing enzyme, I-CsmI, that is encoded in the intronic ORF of a group I intron located in the mitochondrial COB gene of the unicellular green alga Chlamydomonas smithii. We successfully expressed and purified three types of N-terminally truncated I-CsmI polypeptides, and assayed the efficiency of cleavage for 81 substrates containing single nucleotide substitutions. We found a slight but significant tendency that I-CsmI cleaves substrates containing a silent or tolerated amino acid change more efficiently than nonsilent or nontolerated ones. The published recognition properties of I-SpomI, I-ScaI, and I SceII were reconsidered from this point of view, and we detected proficient adaptation of I-SpomI, I-ScaI, and I-SceII for target site sequence degeneracy. Based on the results described above, we propose that intronic homing enzymes are adapted to cleave sequences that might appear at the target region in various species, however, such adaptation becomes less prominent in proportion to the time elapsed after intron invasion into a new host. PMID- 15885099 TI - ERS1 encodes a functional homologue of the human lysosomal cystine transporter. AB - Cystinosis is a lysosomal storage disease caused by an accumulation of insoluble cystine in the lumen of the lysosome. CTNS encodes the lysosomal cystine transporter, mutations in which manifest as a range of disorders and are the most common cause of inherited renal Fanconi syndrome. Cystinosin, the CTNS product, is highly conserved among mammals. Here we show that the yeast Ers1 protein and cystinosin are functional orthologues, despite sharing only limited sequence homology. Ers1 is a vacuolar protein whose loss of function results in growth sensitivity to hygromycin B. This phenotype can be complemented by the human CTNS gene but not by mutant ctns alleles that were previously identified in cystinosis patients. A genetic screen for multicopy suppressors of an ers1Delta yeast strain identified a novel gene, MEH1, which is implicated in regulating Ers1 function. Meh1 localizes to the vacuolar membrane and loss of MEH1 results in a defect in vacuolar acidification, suggesting that the vacuolar environment is critical for normal ERS1 function. This genetic system has also led us to identify Gtr1 as an Meh1 interacting protein. Like Meh1 and Ers1, Gtr1 associates with vacuolar membranes in an Meh1-dependent manner. These results demonstrate the utility of yeast as a model system for the study of CTNS and vacuolar function. PMID- 15885100 TI - Phosphatidylinositol 3,4,5-trisphosphate modulation in SHIP2-deficient mouse embryonic fibroblasts. AB - SHIP2, the ubiquitous SH2 domain containing inositol 5-phosphatase, includes a series of protein interacting domains and has the ability to dephosphorylate phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P(3)]in vitro. The present study, which was undertaken to evaluate the impact of SHIP2 on PtdIns(3,4,5)P(3) levels, was performed in a mouse embryonic fibroblast (MEF) model using SHIP2 deficient (-/-) MEF cells derived from knockout mice. PtdIns(3,4,5)P(3) was upregulated in serum stimulated -/- MEF cells as compared to +/+ MEF cells. Although the absence of SHIP2 had no effect on basal PtdIns(3,4,5)P(3) levels, we show here that this lipid was significantly upregulated in SHIP2 -/- cells but only after short-term (i.e. 5-10 min) incubation with serum. The difference in PtdIns(3,4,5)P(3) levels in heterozygous fibroblast cells was intermediate between the +/+ and the -/- cells. In our model, insulin-like growth factor-1 stimulation did not show this upregulation. Serum stimulated phosphoinositide 3 kinase (PI 3-kinase) activity appeared to be comparable between +/+ and -/- cells. Moreover, protein kinase B, but not mitogen activated protein kinase activity, was also potentiated in SHIP2 deficient cells stimulated by serum. The upregulation of protein kinase B activity in serum stimulated cells was totally reversed in the presence of the PI 3-kinase inhibitor LY-294002, in both +/+ and /- cells. Altogether, these data establish a link between SHIP2 and the acute control of PtdIns(3,4,5)P(3) levels in intact cells. PMID- 15885101 TI - Dual role of oxygen during lipoxygenase reactions. AB - Studying the oxygenation kinetics of (19R/S,5Z,8Z,11Z,14Z)-19-hydroxyeicosa 5,8,11,14-tetraenoic acid (19-OH-AA) by rabbit 15-lipoxygenase-1 we observed a pronounced oxygen dependence of the reaction rate, which was not apparent with arachidonic acid as substrate. Moreover, we found that peroxide-dependent activation of the lipoxygenase depended strongly on the oxygen concentration. These data can be described with a kinetic model that extends previous schemes of the lipoxygenase reaction in three essential aspects: (a) the product of 19-OH-AA oxygenation is a less effective lipoxygenase activator than (13S,9Z,11E)-13 hydroperoxyoctadeca-9,11-dienoic acid; (b) molecular dioxygen serves not only as a lipoxygenase substrate, but also impacts peroxide-dependent enzyme activation; (c) there is a leakage of radical intermediates from the catalytic cycle, which leads to the formation of inactive ferrous lipoxygenase. This enzyme inactivation can be reversed by another round of peroxide-dependent activation. Taken together our data indicate that both peroxide activation and the oxygen affinity of lipoxygenases depend strongly on the chemistry of the lipid substrate. These findings are of biological relevance as variations of the reaction conditions may turn the lipoxygenase reaction into an efficient source of free radicals. PMID- 15885102 TI - Production and characterization of a noncytotoxic deletion variant of the Aspergillus fumigatus allergen Aspf1 displaying reduced IgE binding. AB - Aspergillus fumigatus is responsible for many allergic respiratory diseases, the most notable of which - due to its severity - is allergic bronchopulmonary aspergillosis. Aspf1 is a major allergen of this fungus: this 149-amino acid protein belongs to the ribotoxin family, whose best characterized member is alpha sarcin (EC 3.1.27.10). The proteins of this group are cytotoxic ribonucleases that degrade a unique bond in ribosomal RNA impairing protein biosynthesis. Aspf1 and its deletion mutant Aspf1Delta(7-22) have been produced as recombinant proteins; the deleted region corresponds to an exposed beta-hairpin. The conformation of these two proteins has been studied by CD and fluorescence spectroscopy. Their enzymatic activity and cytotoxicity against human rhabdomyosarcoma cells was also measured and their allergenic properties have been studied by using 58 individual sera of patients sensitized to Aspergillus. Aspf1Delta(7-22) lacks cytotoxicity and shows a remarkably reduced IgE reactivity. From these studies it can be concluded that the deleted beta-hairpin is involved in ribosome recognition and is a significant allergenic region. PMID- 15885103 TI - Differential expression of endogenous sialidases of human monocytes during cellular differentiation into macrophages. AB - Sialidases are enzymes that influence cellular activity by removing terminal sialic acid from glycolipids and glycoproteins. Four genetically distinct sialidases have been identified in mammalian cells. In this study, we demonstrate that three of these sialidases, lysosomal Neu1 and Neu4 and plasma membrane associated Neu3, are expressed in human monocytes. When measured using the artificial substrate 2'-(4-methylumbelliferyl)-alpha-d-N-acetylneuraminic acid (4 MU-NANA), sialidase activity of monocytes increased up to 14-fold per milligram of total protein after cells had differentiated into macrophages. In these same cells, the specific activity of other cellular proteins (e.g. beta-galactosidase, cathepsin A and alkaline phosphatase) increased only two- to fourfold during differentiation of monocytes. Sialidase activity measured with 4-MU-NANA resulted from increased expression of Neu1, as removal of Neu1 from the cell lysate by immunoprecipitation eliminated more than 99% of detectable sialidase activity. When exogenous mixed bovine gangliosides were used as substrates, there was a twofold increase in sialidase activity per milligram of total protein in monocyte derived macrophages in comparison to monocytes. The increased activity measured with mixed gangliosides was not affected by removal of Neu1, suggesting that the expression of a sialidase other than Neu1 was present in macrophages. The amount of Neu1 and Neu3 RNAs detected by real time RT-PCR increased as monocytes differentiated into macrophages, whereas the amount of Neu4 RNA decreased. No RNA encoding the cytosolic sialidase (Neu2) was detected in monocytes or macrophages. Western blot analysis using specific antibodies showed that the amount of Neu1 and Neu3 proteins increased during monocyte differentiation. Thus, the differentiation of monocytes into macrophages is associated with regulation of the expression of at least three distinct cellular sialidases, with specific up regulation of the enzyme activity of only Neu1. PMID- 15885104 TI - Affinity of S100A1 protein for calcium increases dramatically upon glutathionylation. AB - S100A1 is a typical representative of a group of EF-hand calcium-binding proteins known as the S100 family. The protein is composed of two alpha subunits, each containing two calcium-binding loops (N and C). At physiological pH (7.2) and NaCl concentration (100 mm), we determined the microscopic binding constants of calcium to S100A1 by analysing the Ca(2+)-titration curves of Trp90 fluorescence for both the native protein and its Glu32 --> Gln mutant with an inactive N-loop. Using a chelator method, we also determined the calcium-binding constant for the S100A1 Glu73 --> Gln mutant with an inactive C-loop. The protein binds four calcium ions in a noncooperative way with binding constants of K(1) =4 +/- 2 x 10(3) m(-1) (C-loops) and K(2) approximately 10(2) m(-1) (N-loops). Only when both loops are saturated with calcium does the protein change its global conformation, exposing to the solvent hydrophobic patches, which can be detected by 2-p-toluidinylnaphthalene-6-sulfonic acid - a fluorescent probe of protein surface hydrophobicity. S-Glutathionylation of the single cysteine residue (85) of the alpha subunits leads to a 10-fold increase in the affinity of the protein C-loops for calcium and an enormous - four orders of magnitude - increase in the calcium-binding constants of its N-loops, owing to a cooperativity effect corresponding to DeltaDeltaG = -6 +/- 1 kcal.mol(-1). A similar effect is observed upon formation of the mixed disulfide with cysteine and 2 mercaptoethanol. The glutathionylated protein binds TRTK-12 peptide in a calcium dependent manner. S100A1 protein can act, therefore, as a linker between the calcium and redox signalling pathways. PMID- 15885105 TI - Quantitative interdependence of coeffectors, CcpA and cre in carbon catabolite regulation of Bacillus subtilis. AB - The phosphoproteins HPrSerP and CrhP are the main effectors for CcpA-mediated carbon catabolite regulation (CCR) in Bacillus subtilis. Complexes of CcpA with HPrSerP or CrhP regulate genes by binding to the catabolite responsive elements (cre). We present a quantitative analysis of HPrSerP and CrhP interaction with CcpA by surface plasmon resonance (SPR) revealing small and similar equilibrium constants of 4.8 +/- 0.4 microm for HPrSerP-CcpA and 19.1 +/- 2.5 microm for CrhP CcpA complex dissociation. Forty millimolar fructose-1,6-bisphosphate (FBP) or glucose-6-phosphate (Glc6-P) increases the affinity of HPrSerP to CcpA at least twofold, but have no effect on CrhP-CcpA binding. Saturation of binding of CcpA to cre as studied by fluorescence and SPR is dependent on 50 microm of HPrSerP or > 200 microm CrhP. The rate constants of HPrSerP-CcpA-cre complex formation are k(a) = 3 +/- 1 x 10(6) m(-1).s(-1) and k(d) = 2.0 +/- 0.4 x 10(-3).s(-1), resulting in a K(D) of 0.6 +/- 0.3 nm. FBP and Glc6-P stimulate CcpA-HPrSerP but not CcpA-CrhP binding to cre. Maximal HPrSerP-CcpA-cre complex formation in the presence of 10 mm FBP requires about 10-fold less HPrSerP. These data suggest a specific role for FBP and Glc6-P in enhancing only HPrSerP-mediated CCR. PMID- 15885106 TI - Calpain 1-titin interactions concentrate calpain 1 in the Z-band edges and in the N2-line region within the skeletal myofibril. AB - Calpain 1, a ubiquitous calcium-dependent intracellular protease, was recently found in a tight association with myofibrils in skeletal muscle tissue [Delgado EF, Geesink GH, Marchello JA, Goll DE & Koohmaraie M (2001) J Anim Sci79, 2097 2107). Our immunofluorescence and immunoelectron microscopy investigations restrain the protease location at the periphery of the Z-band and at the midpoint of the I-band. Furthermore, calpain 1 is found to localize in myofibril fractures, described as proteolysis sites, in postmortem bovine skeletal red muscles, near the calcium deposits located at the N1 and N2 level. This in situ localization of calpain 1 is substantiated by binding assays with two titin regions covering the I-band region: a native fragment of 150 kDa (identified by mass spectrometry) that includes the N-terminal Z8-I5 region and the N1-line region of titin, and an 800 kDa fragment external to the N1 line that bears the PEVK/N2 region. These two titin fragments are shown to tightly bind calpain 1 in the presence of CaCl(2) and E64, a calpain inhibitor. In the absence of E64, they are cleaved by calpain 1. We conclude that titin affords binding sites to calpain 1, which concentrates the protease in the regions restrained by the Z-band edge and the N1-line as well as at the N2-line level, two sarcomeric regions where early postmortem proteolysis is detected. PMID- 15885107 TI - A strategy for the generation of specific human antibodies by directed evolution and phage display. An example of a single-chain antibody fragment that neutralizes a major component of scorpion venom. AB - This study describes the construction of a library of single-chain antibody fragments (scFvs) from a single human donor by individual amplification of all heavy and light variable domains (1.1 x 10(8) recombinants). The library was panned using the phage display technique, which allowed selection of specific scFvs (3F and C1) capable of recognizing Cn2, the major toxic component of Centruroides noxius scorpion venom. The scFv 3F was matured in vitro by three cycles of directed evolution. The use of stringent conditions in the third cycle allowed the selection of several improved clones. The best scFv obtained (6009F) was improved in terms of its affinity by 446-fold, from 183 nm (3F) to 410 pm. This scFv 6009F was able to neutralize 2 LD(50) of Cn2 toxin when a 1 : 10 molar ratio of toxin-to-antibody fragment was used. It was also able to neutralize 2 LD(50) of the whole venom. These results pave the way for the future generation of recombinant human antivenoms. PMID- 15885108 TI - The binding of foot-and-mouth disease virus leader proteinase to eIF4GI involves conserved ionic interactions. AB - The leader proteinase (L(pro)) of foot-and-mouth disease virus (FMDV) initially cleaves itself from the polyprotein. Subsequently, L(pro) cleaves the host proteins eukaryotic initiation factor (eIF) 4GI and 4GII. This prevents protein synthesis from capped cellular mRNAs; the viral RNA is still translated, initiating from an internal ribosome entry site. L(pro) cleaves eIF4GI between residues G674 and R675. We showed previously, however, that L(pro) binds to residues 640-669 of eIF4GI. Binding was substantially improved when the eIF4GI fragment contained the eIF4E binding site and eIF4E was present in the binding assay. L(pro) interacts with eIF4GI via residue C133 and residues 183-195 of the C-terminal extension. This binding domain lies about 25 A from the active site. Here, we examined the binding of L(pro) to eIF4GI fragments generated by in vitro translation to narrow the binding site down to residues 645-657 of human eIF4GI. Comparison of these amino acids with those in human eIF4GII as well as with sequences of eIF4GI from other organisms allowed us to identify two conserved basic residues (K646 and R650). Mutation of these residues was severely detrimental to L(pro) binding. Similarly, comparison of the sequence between residues 183 and 195 of L(pro) with those of other FMDV serotypes and equine rhinitis A virus showed that acidic residues D184 and E186 were highly conserved. Substitution of these residues in L(pro) significantly reduced eIF4GI binding and cleavage without affecting self-processing. Thus, FMDV L(pro) has evolved a domain that specifically recognizes a host cell protein. PMID- 15885109 TI - Special issue dedicated to Victor J. Hruby and the advancement of peptide chemistry, biology and pharmaceutical sciences. PMID- 15885110 TI - Hail to the editor-in-chief: an appreciation of Victor J. Hruby, outstanding scientist-mentor-leader in peptide research. PMID- 15885111 TI - Combined solid-phase/solution synthesis of large ribonuclease A C-terminal peptides containing a non-natural proline analog. AB - Three large peptides corresponding to the 65-124 (60-mer), 72-124 (53-mer), and 77-124 (48-mer) sequence of bovine pancreatic ribonuclease A (RNase A) were assembled from either two or three shorter protected peptide fragments by chemical coupling in solution. The fragments were synthesized manually by 9 fluorenylmethyloxycarbonyl (Fmoc)-based solid-phase peptide chemistry in plastic syringes, and subsequently purified by normal-phase high-performance liquid chromatography on a silica gel column. The main aim of this work was to incorporate sterically hindered l-5,5-dimethylproline (dmP) as a substitute for Pro(93) into the sequence of RNase A in order to constrain the -Tyr(92)-Pro(93)- peptide group to a single cis-conformation. PMID- 15885112 TI - Evaluation of geranylazide and farnesylazide diphosphate for incorporation of prenylazides into a CAAX box-containing peptide using protein farnesyltransferase. AB - Protein farnesyltransferase (PFTase) catalyzes the attachment of a geranylazide (C10) or farnesylazide (C15) moiety from the corresponding prenyldiphosphates to a model peptide substrate, N-dansyl-Gly-Cys-Val-Ile-Ala-OH. The rates of incorporation for these two substrate analogs are comparable and approximately twofold lower than that using the natural substrate farnesyl diphosphate (FPP). Reaction of N-dansyl-Gly-Cys(S-farnesylazide)-Val-Ile-Ala-OH with 2 diphenylphosphanylbenzoic acid methyl ester then gives a stable alkoxy-imidate linked product. This result suggests future generations whereby azide groups introduced using this enzymatic approach are functionalized using a broad range of azide-reactive reagents. Thus, chemistry has been developed that could be used to achieve highly specific peptide and protein modification. The farnesylazide analog may be useful in certain biological studies, whereas the geranylazide group may be more useful for general protein modification and immobilization. PMID- 15885113 TI - Monomeric and dimeric states exhibited by the kinesin-related motor protein KIF1A. AB - KIF1A, a kinesin-related motor protein that transports pre-synaptic vesicles in neurons, was originally presumed to translocate along microtubules (MT) as a monomer. Protein structure predictions from its amino acid sequence failed to identify the long coiled-coil domains typical of kinesins, which led researchers to believe it does not oligomerize into the canonical kinesin dimer. However, mounting evidence using recombinant chimeric protein indicates that KIF1A, like conventional kinesin, requires dimerization for fast, unidirectional processive movement along MTs. Because these studies are somewhat indirect, we wished to test the oligomerization state of native KIF1A, and to compare that to full length recombinant protein. We have performed hydrodynamic analyses to determine the molecular weights of the respective complexes. Our results indicate that most native KIF1A is soluble and indeed monomeric, but recombinant KIF1A is a dimer. MT-binding studies also showed that native KIF1A did not bind to MTs in either the presence of AMP-PNP, apyrase, or adenosine triphosphate (ATP), but recombinant KIF1A bound to MTs most stably in the presence of ATP, indicating very different motor functional states. To further characterize KIF1A's dimerization potential, we prepared peptides corresponding to the neck domains of MmKIF1A and CeUnc104, and by circular dichroism spectroscopy compared these peptides for their ability to form coiled-coils. Interestingly, both MmKIF1A and CeUnc104 neck peptides formed homodimeric coiled-coils, with the MmKIF1A neck coiled-coil exhibiting the greater stability. Collectively, from our data and from previous studies, we predict that native KIF1A can exist as both an inactive monomer and an active homodimer formed in part through its neck coiled-coil domain. PMID- 15885114 TI - Abbreviated nomenclature for cyclic and branched homo- and hetero-detic peptides. AB - Amino acid sequences and linear or head-to-tail cyclopeptides can be represented conveniently in one-line text formulae using the three-letter symbols. However, other - but nonetheless important - topologies of peptides are 'side chain-to head (or tail)', 'backbone-to-backbone', 'side chain-to-side chain' cyclopeptides, 'side chain-to-side chain' connected peptide strands, and branched peptides (like peptide dendrimers). In general, such structures cannot be described using the three-letter symbols in one-line text: a chemical structure editor is required for symbolic representations according to the IUPAC-IUBMB recommendations. The aim of this contribution is to offer an unambiguous and general nomenclature system that enables researchers to represent all cyclic and branched homo- and hetero-detic peptides in a coherent manner in one-line text - as long as their as constituents can be represented in (three)-letter codes. The application of this new nomenclature would overcome the existing difficulties and provide a way to express complex situations in the shortest way in order to highlight more clearly the salient points in a given scientific communication. PMID- 15885115 TI - Type and location of fluorescent probes incorporated into the potent mu-opioid peptide [Dmt]DALDA affect potency, receptor selectivity and intrinsic efficacy. AB - The dermorphin-derived tetrapeptide H-Dmt-d-Arg-Phe-Lys-NH(2) (Dmt = 2',6' dimethyltyrosine) ([Dmt(1)]DALDA) is a highly potent and selective mu-opioid agonist capable of crossing the blood-brain barrier and producing a potent, centrally mediated analgesic effect when given systemically. For the purpose of biodistribution studies by fluorescence techniques, [Dmt(1)]DALDA analogues containing various fluorescent labels [dansyl, anthraniloyl (atn), fluorescein, or 6-dimethylamino-2'-naphthoyl] in several different locations of the peptide were synthesized and characterized in vitro in the guinea-pig ileum and mouse vas deferens assays, and in mu-, delta- and kappa-opioid receptor-binding assays. The analogues showed various degrees of mu receptor-binding selectivity, but all of them were less mu-selective than the [Dmt(1)]DALDA parent peptide. Most analogues retained potent, full mu-agonist activity, except for one with fluorescein attached at the C-terminus (3a) (partial mu-agonist) and one containing beta-(6' dimethylamino-2'-naphthoyl)alanine (aladan) in place of Phe(3) (4) (mu- and kappa antagonist). The obtained data indicate that the receptor-binding affinity, receptor selectivity and intrinsic efficacy of the prepared analogues vary very significantly, depending on the type of fluorescent label used and on its location in the peptide. The results suggest that the biological activity profile of fluorescence-labeled peptide analogues should always be carefully determined prior to their use in biodistribution studies or other studies. One of the analogues containing the atn group (2a) proved highly useful in a study of cellular uptake and intracellular distribution by confocal laser scanning microscopy. PMID- 15885116 TI - A topographically and conformationally constrained, spin-labeled, alpha-amino acid: crystallographic characterization in peptides. AB - 2,2,6,6-Tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) is a topographically and conformationally restricted, nitroxide containing, C(alpha) tetrasubstituted alpha-amino acid. Here, we describe the molecular and crystal structures, as determined by X-ray diffraction analyses, of a TOAC terminally protected derivative, the cyclic dipeptide c(TOAC)(2).1,1,1,3,3,3 hexafluoropropan-2-ol (HFIP) solvate, and five TOAC-containing, terminally protected, linear peptides ranging in length from tetra- to hepta-peptides. Incipient and fully developed, regular or distorted 3(10)-helical structures are formed by the linear peptides. A detailed discussion on the average geometry and preferred conformation for the TOAC piperidine ring is also reported. The X-ray diffraction structure of an intramolecularly cyclized side product resulting from a C-activated TOAC residue has also been determined. PMID- 15885117 TI - Fatty acyl moieties: improving Pro-rich peptide uptake inside HeLa cells. AB - In the field of drug delivery there has been a continuous study of powerful delivery systems to aid non permeable drugs in reaching their intracellular target. Among the systems explored are cell penetrating peptides (CPPs), which first garnered interest a decade ago when the interesting translocation properties of the pioneer CPPs Tat and Antp were described. A new family of CPPs has recently been described as non cytotoxic Pro-rich vectors with favorable profiles for internalization in HeLa cells. Fatty acyl moieties that can tune a peptide's interaction with the lipophilic environment of a cell membrane have been incorporated into the Pro-rich sequence. Improvements in cellular uptake of peptides modified with fatty acyl groups, as studied by confocal microscopy and flow cytometry, as well as the results obtained by the interaction of these peptides with a model dioleoylphosphatidylcholine (DOPC) membrane and transmission electron microscopy (TEM), illustrate the importance of the fatty acyl moieties for efficient internalization. PMID- 15885118 TI - Therapeutic peptidomimetic strategies for autoimmune diseases: costimulation blockade. AB - Cognate interactions between immune effector cells and antigen-presenting cells (APCs) govern immune responses. Specific signals occur between the T-cell receptor peptide and APCs and nonspecific signals between pairs of costimulatory molecules. Costimulation signals are required for full T-cell activation and are assumed to regulate T-cell responses as well as other aspects of the immune system. As new discoveries are made, it is becoming clear how important these costimulation interactions are for immune responses. Costimulation requirements for T-cell regulation have been extensively studied as a way to control many autoimmune diseases and downregulate inflammatory reactions. The CD28:B7 and the CD40:CD40L families of molecules are considered to be critical costimulatory molecules and have been studied extensively. Blocking the interaction between these molecules results in a state of immune unresponsiveness termed 'anergy'. Several different strategies for blockade of these interactions are explored including monoclonal antibodies (mAbs), Fab fragments, chimeric, and/or fusion proteins. We developed novel, immune-specific approaches that interfere with these interactions. Using experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis mediated by central nervous system (CNS) specific T-cells, we developed a multi-targeted approach that utilizes peptides for blockade of costimulatory molecules. We designed blocking peptide mimics that retain the functional binding area of the parent protein while reducing the overall size and are thus capable of blocking signal transduction. In this paper, we review the role of costimulatory molecules in autoimmune diseases, two of the most well-studied costimulatory pathways (CD28/CTLA-4:B7 and CD40:CD40L), and the advantages of peptidomimetic approaches. We present data showing the ability of peptide mimics of costimulatory molecules to suppress autoimmune disease and propose a mechanism for disease suppression. PMID- 15885119 TI - Peptides and the development of double- and triple-resonance solid-state NMR of aligned samples. AB - Peptides have been instrumental in the development of solid-state nuclear magnetic resonance (NMR) spectroscopy, and their roles in the development of solid-state NMR of aligned samples is reviewed. In particular, the roles of synthetic peptides in the development of triple-resonance methods are described. Recent developments of pulse sequences and NMR probes for triple-resonance NMR of aligned samples are presented. PMID- 15885120 TI - Immune responses that adapt the intestinal mucosa to commensal intestinal bacteria. AB - Animals contain an enormous load of non-pathogenic bacteria in the lower intestine, which exploit an environment with a stable temperature and abundant carbon sources. Our load of bacteria outnumbers our own cells. In order to survive with such a high number of organisms in very close proximity to host tissues the intestinal mucosa and its immune system is highly adapted. Mucosal immune responses are induced by small numbers of live commensal organisms penetrating the Peyer's patches and persisting in dendritic cells (DC). These DC can induce immunoglobulin A+ (IgA+) B cells, which recirculate through the lymph and bloodstream to populate the lamina propria and secrete protective IgA. Because DC loaded with commensal bacteria do not penetrate further than the mesenteric lymph nodes, immune induction to commensals is confined to the mucosa, allowing strong mucosal immune responses to be induced whilst the systemic immune system remains relatively ignorant of these organisms. PMID- 15885121 TI - Building better T-cell-inducing malaria vaccines. AB - Since malaria continues to account for millions of deaths annually in endemic regions, the development of an effective vaccine remains highly desirable. The life cycle of malaria poses a number of challenges to the immune response since phases of the cycle express varying antigen profiles and have different locations, thus requiring differing antigenic targets and effector mechanisms. To confer sterile immunity, a vaccine would have to target the pre-erythrocytic stages of infection. Since at this stage the parasite is hidden within liver cells, the host defence predominantly requires cell-mediated immunity, chiefly T cells, to eliminate infected hepatocytes. The development of such vaccines has progressed from irradiated sporozoites, through recombinant proteins, to recombinant DNA and viral vectors. Some of the experimental vaccination regimens that explore various combinations of vaccines for priming and boosting, together with numbers of vaccinations, interval between them, and the vaccination site, are revealing strong immunogenicity and evidence of efficacy in human challenge studies and in field trials. Such approaches should lead to deployable vaccines that protect against malarial disease. PMID- 15885122 TI - The negative regulatory function of the lymphocyte-activation gene-3 co-receptor (CD223) on human T cells. AB - Accumulating evidence indicates that the CD4 homologue lymphocyte activation gene 3 (LAG-3) plays a down-regulatory role on T-cell responses. However, the role of LAG-3/major histocompatibility complex (MHC) class II interactions on primary human T-cell responses, as well as the mechanism by which down-regulation occurs, are not clear. Here, we show that LAG-3 colocalized with CD3, CD4 or CD8 in areas of cholesterol-rich raft aggregation during this primary response, as well as in the clustered raft region formed between T cells and antibody-coated beads. Addition of a blocking LAG-3-specific monoclonal antibody to both CD4 and CD8 primary resting T cells activated under conditions of antigen-presenting cell driven stimulation and low antigen concentrations augments CD69 activation antigen expression, T-cell expansion and T helper 1 (Th1, but not Th2) cytokine production. Blocking LAG-3/MHC class II interactions leads to an increase in the number of cells entering division at these low concentrations of antigen and to more rounds of divisions with an accumulation of cells in the S-phase of the cell cycle. These results indicate that LAG-3 signalling inhibits early events in primary activation of human CD4 and CD8 T cells and further support a role for LAG-3 signalling in regulating the expansion of activated effector or memory T cells, either directly or indirectly through Treg suppressor activity. PMID- 15885123 TI - Differential expression of regulator of G-protein signalling transcripts and in vivo migration of CD4+ naive and regulatory T cells. AB - The immune response of T lymphocytes to pathogens is initiated in draining secondary lymphoid organs, and activated cells then migrate to the site of infection. Thus, control of naive and regulatory CD4+ T-cell migration is crucial; however, it is poorly understood in physiological and pathological conditions. We found that CD4+ subpopulations displayed characteristic regulator of G-protein signalling (RGS) gene expression profiles. Regulatory T cells express higher levels of RGS1, RGS9 and RGS16 than naive cells. These genes are up-regulated upon cell activation and their level of expression correlates with in vivo cell migration. Using parabiosis, we showed that regulatory T lymphocytes migrate less than naive T cells and that migrant naive T cells express even lower RGS levels than their static counterparts. Our results show an inverse correlation between the capacity to migrate and the levels of RGS1, RGS9 and RGS16 for both naive and regulatory T cells. Taken together, these results suggest a role for RGS molecules in chemokine-induced lymphocyte migration and demonstrate the peculiarity of regulatory T cells in terms of phenotype and migration ability, providing new insights into their function. PMID- 15885124 TI - Monoclonal antibodies that identify the CD3 molecules expressed specifically at the surface of porcine gammadelta-T cells. AB - The CD3 antigen is a surface structure associated with the T-cell receptor (TCR) to form a complex involved in antigen recognition and signal transduction. Reports on the structures of the CD3 molecules associated with alphabeta- and gammadelta-TCR have been contradictory. To investigate this issue, we raised a panel of monoclonal antibodies (mAb) against purified porcine CD3 molecules. Unlike the conventional anti-CD3, these mAb reacted specifically with peripheral gammadelta-T cells, but not with alphabeta-T cells. Immunoprecipitation showed that the antibody recognized a subset of CD3 molecules that were associated with gammadelta-TCR. Also unlike the conventional anti-CD3, these mAb, though directed at two different epitope groups, failed to induce antigenic modulation, T-cell proliferation and CD3-redirected cytotoxicity. Taken together, these results suggest that there are differences in the antigenicity, signal transduction potentials and probably structural differences between the CD3 molecules expressed at the surface of alphabeta- and gammadelta-T cells. PMID- 15885125 TI - The Vgamma2/Vdelta2 T-cell repertoire in Macaca fascicularis: functional responses to phosphoantigen stimulation by the Vgamma2/Jgamma1.2 subset. AB - Circulating Vgamma2/Vdelta2 T cells in human and non-human primates respond to small molecular weight non-peptidic phosphoantigens in a major histocompatibility complex (MHC)-unrestricted manner. These responses are encoded by the Vgamma2/Jgamma1.2 chain of the T-cell receptor and are positively selected during early development to create a biased repertoire in adults. We characterized the Vgamma2 chain in cynomolgus macaques (Macaca fascicularis) to develop a non-human primate model for studying the effects of infection and therapy on the circulating Vgamma2/Vdelta2 T-cell subset. The cynomolgus macaque Vgamma2 chain was highly homologous to the Vgamma2 chain from human beings and rhesus macaques (Macaca mulatta), though we noted conserved substitutions in critical residues within the CDR3 for both macaque species. Despite these substitutions, Vgamma2/Vdelta2+ T cells from cynomolgus monkeys exhibited polyclonal responses to two different phosphoantigens. Proliferative responses were observed with both isopentenylpyrophosphate and alendronate, but stronger interferon-gamma secretory responses were observed with isopentenylpyrophosphate. In vitro stimulation and expansion led to selective outgrowth of the Vgamma2/Jgamma1.2 subset, with a marked shift in the Vgamma2 spectratype. As a result of the less biased starting repertoire for Vgamma2, the cynomolgus macaque constitutes a sensitive model for examining the effects of in vitro or in vivo treatments on the Vgamma2/Vdelta2 T cell population. Our studies establish the value of cynomolgus macaques as a model for Vgamma2/Vdelta2 T-cell responses to non-peptidic antigens, and further evidence the remarkable evolutionary conservation of this unusual, phosphoantigen responsive T-cell subset that is found only in primate species. PMID- 15885126 TI - Delineation of multiple subpopulations of natural killer cells in rhesus macaques. AB - Natural killer (NK) cells in rhesus macaques have been variably defined as CD3- CD16+ or CD3- CD8+, although only limited efforts have been made to validate these definitions rigorously. To better understand the role of NK cells in macaque disease models, we undertook a multiparameter analysis of macaque NK cells employing four-colour flow cytometry and a panel of lineage-specific and non-lineage-specific lymphocyte markers. Using this approach, we identified two distinct populations of candidate NK cells: a major CD8bright CD16+ population and a minor CD8bright CD16- population. Further analysis of the major and minor NK cell populations revealed the expression of multiple markers characteristic of NK cells, including CD2, CD7, CD16, CD161, NKG2A and granzyme B. In addition, a CD56+ subset of cells within the minor rhesus NK population was identified which expressed chemokine and lymph node homing receptors similar to those expressed by the CD56bright NK cell population identified in humans. Cytolytic assays confirmed that the phenotypically defined rhesus NK cells lysed NK-susceptible target cells. Our observations support the existence of several distinct subpopulations of rhesus macaque NK cells, which have significant phenotypic and functional similarities to their human counterparts. These improved immunophenotypic definitions of macaque NK cells should facilitate future analysis of innate immune responses in rhesus macaques and the role of NK cells in AIDS pathogenesis in Simian immunodeficiency virus (SIV)-infected macaques. PMID- 15885127 TI - Synergistic antibody induction by antigen-CD40 ligand fusion protein as improved immunogen. AB - Full scale B-cell activation requires not only B-cell receptor (BCR) engagement with antigen, but also costimulatory signals provided by T helper cells through the CD40-CD40 ligand (CD40L) interaction. It is hypothesized that a fusion protein of an antigen and soluble CD40L (CD40LT) would selectively target the costimulation to antigen-specific B cells, leading to synergy in the antibody response. This hypothesis was investigated by fusing green fluorescence protein (GFP), a generic antigen, with mouse CD40LT. Studies revealed that immunization in mice with the plasmid encoding GFP-CD40LT fusion protein led to synergistic induction of GFP-specific antibodies, while control plasmid(s) for GFP, CD40LT, or GFP plus CD40LT did not. Immunization with a single dose of the fusion protein also provoked a vigorous GFP-specific immunoglobulin G1 antibody response, but not other antibody isotypes. These results suggest that GFP-CD40LT fusion protein induces a GFP-specific B-cell activation and antibody response in an antigen guided fashion. The potential application of this novel strategy in vaccine development is discussed. PMID- 15885128 TI - CpG motif acts as a 'danger signal' and provides a T helper type 1-biased microenvironment for DNA vaccination. AB - The method of delivering a DNA vaccine can influence the type of immune response induced by the vaccine. Application of a DNA vaccine by gene gun typically induces a T helper type 2 (Th2)-type reaction, whereas needle inoculation triggers a Th1 response. In the present study, we found that physical trauma, gold-particle bombardment and the CpG motif can act as 'danger signals' that recruit inflammatory cells to damaged tissues. Analysis of the cytokine profiles of draining lymph nodes or lymph-node-derived mononuclear cells from different groups by real-time reverse transcription-polymerase chain reaction revealed that, while gene-gun-bombardment induced a Th2-type cytokine microenvironment with increased interleukin-4 (IL-4) and IL-10 mRNA expression and almost no increase in IL-12 and interferon-gamma mRNA expression in draining lymph nodes, intradermal injection as well as subcutaneous injection of muscle induced the opposite. We further studied whether the addition of the CpG motif can switch the Th2-type cytokine microenvironment produced by gene-gun bombardment in draining lymph nodes. Results showed that the addition of the CpG motif can increase IL-12 mRNA expression in draining lymph nodes whether induced by intradermal injection, intramuscular injection, or gene-gun bombardment. These data suggest that delivery of the CpG motif induced a Th1-biased microenvironment in draining lymph nodes. Taken together, the CpG motif can act as a 'danger signal' and Th1 immune response enhancer in DNA vaccination. These results may help to explain the mechanism of different types of immune response induced by DNA vaccines delivered by different routes and facilitate the application of DNA vaccines. PMID- 15885129 TI - Comparative cell signalling activity of ultrapure recombinant chaperonin 60 proteins from prokaryotes and eukaryotes. AB - Heat-shock protein (hsp)60/chaperonin 60 is a potent immunogen which has recently been claimed to have cell-signalling actions upon myeloid and vascular endothelial cells. The literature is controversial with different chaperonin 60 proteins producing different patterns of cellular activation and the ever-present criticism that activity is the result of bacterial contaminants. To clarify the situation we have cloned, expressed and purified to homogeneity the chaperonin 60 proteins from Chlamydia pneumoniae, Helicobacter pylori and the human mitochondrion. These highly purified proteins were compared for their ability to stimulate human peripheral blood mononuclear cell (PBMC) cytokine synthesis and vascular endothelial cell adhesion protein expression. In spite of their significant sequence homology, the H. pylori protein was the most potent PBMC activator with the human protein the least potent. PBMC activation by C. pneumoniae and human, but not H. pylori, chaperonin 60 was blocked by antibody neutralization of Toll-like receptor-4. The C. pneumoniae chaperonin 60 was the most potent endothelial cell activator, with the human protein being significantly less active than bacterial chaperonin 60 proteins. These results have implications for the role of chaperonin 60 proteins as pathological factors in autoimmune and cardiovascular disease, and raise the possibility that each of these proteins may result in different pathological effects in such diseases. PMID- 15885130 TI - Human CD8+ T cells specific for influenza A virus M1 display broad expression of maturation-associated phenotypic markers and chemokine receptors. AB - To define the role of memory T cells in a non-persistent viral infection, we have delineated the phenotype of memory CD8+ T cells specific for influenza A virus (FluA; matrix protein M158-66) based on the expression of several memory/effector lineage markers and relevant chemokine receptors. We found a majority of FluA specific CD8+ T cells expressed CD27 and CD28, and variably expressed CD45RA, CD62L, CD94 and granzyme A. A majority of FluA-specific CD8+ T cells expressed high levels of CXCR3, and moderate levels of CCR5 and CXCR4, whereas a limited proportion expressed CCR7, CCR6 and CXCR5. A phenotypic profile based on these observations showed that there are both immature and mature memory CD8+ T cells specific for FluA. PMID- 15885131 TI - Interleukin-10 production by lung macrophages in CBA xid mutant mice infected with Mycobacterium tuberculosis. AB - Mice on the CBA inbred strain background expressing the well characterized mutation designated xid in the cytoplasmic signalling enzyme Bruton's protein kinase have been previously noted to illustrate shifts in T helper type 1 (Th1)/Th2 immunity which is underlined by an apparent failure to produce the regulatory cytokine interleukin-10. In the current study we examined if this extended to infection with Mycobacterium tuberculosis, which also depends on Th1 immunity. Contrary to expectations, xid mice showed evidence of a transient early susceptibility to pulmonary infection, changes in macrophage morphology, and decreased activation of lung natural killer cells, while showing evidence of substantial IL-10 production and accumulation in lung lesions macrophages, but paradoxically this did not influence the course of the chronic disease. In addition, macrophages from the lungs of xid mice also expressed high levels of CD14. These observations suggest that the xid mutation in cellular signalling has much wider effects on the immune system than previously thought. PMID- 15885132 TI - Increased carrageenan-induced acute lung inflammation in old rats. AB - Ageing is associated with increased susceptibility to lung infections and delayed resolution of pulmonary infiltrates. The purpose of this study was to investigate the effect of age on the onset of carrageenan-induced lung inflammation. When compared with carrageenan-treated young rats (3 months old), old rats (>18 months old) exhibited a preponderance of pleural exudation and polymorphonuclear cell infiltration. Lung myeloperoxidase activity, an index of neutrophil infiltration and activation, was significantly increased in old rats in comparison with young rats. Consistent with the biochemical markers of inflammation, increased lung damage, as assessed by nitrosative stress and lipid peroxidation, was observed in carrageenan-treated old rats. In the lung exudate obtained from old rats, a significant reduction in interleukin-10 (IL-10) was observed, while similar expression of monocyte chemotactic protein-1 was induced, suggesting that a decrease in IL-10 rather than increased chemotaxis may account for the preponderance of the inflammatory cellular infiltrate in old rats. Similar to the in vivo situation, freshly isolated alveolar macrophages obtained from old rats produced less IL-10. This defective IL-10 production could be explained by a reduction in the cAMP-dependent signalling pathway, which mediates IL-10 production. Indeed, we found decreased cAMP-responsive element binding protein (CREB) and phosphorous-CREB (P-CREB) expression in old rats, which may account for reduced IL-10 production in old rats. PMID- 15885133 TI - Functional characterization of T cells in abdominal aortic aneurysms. AB - Abdominal aortic aneurysms (AAA) exhibit features of a chronic inflammatory disorder. The functional attributes of the T cells in AAA tissue are unclear, with little quantitative or functional data. Using a novel, non-enzymatic method to isolate viable cells from AAA tissue, functional properties of AAA T cells were investigated for the first time. Composition and phenotype of AAA T cells was determined by flow cytometry and verified by immunohistochemistry. Tissue mononuclear cells (MNCs) were cultured in the presence of T-cell mitogens, and cell cycle analysis and cytokine production assessed. Typical cell yield was 4.5 x 10(6) cells per gram of AAA tissue. The majority (58.1+/-5.3%) of haematopoietic (CD45+) cells recovered were CD3+ T cells, B cells comprised 41.1+/-5.7%, natural killer cells 7.3+/-2.5%, and macrophages 2%. Freshly isolated T cells were in resting (G1) state, with 25% expressing the activation associated cell surface antigens major histocompatibility complex II and CD25. When stimulated in vitro, a significant proportion entered S and G2 phase of the cell cycle, up-regulated CD25, and secreted tumour necrosis factor-alpha, interferon-gamma, interleukin (IL)-5 and IL-6. Despite patient differences, the composition of the AAA inflammatory infiltrate was remarkably consistent, and when re-stimulated ex-vivo T cells produced a stereotypical cytokine response, consistent with the hypothesis that AAA T cells can promote tissue inflammation by secretion of proinflammatory cytokines, and in addition provide signals for B cell help. PMID- 15885134 TI - Impaired B cell responses to orally administered antigens in lamina propria but not Peyer's patches of Galphai2-deficient mice prior to colitis. AB - Despite numerous studies on the intestinal immune system in patients with inflammatory bowel disease (IBD) and animal models of IBD, very little is known about the immune reactivity of mucosal lymphocytes following oral immunizations under these circumstances. The reactivity of Peyer's patch (PP) and lamina propria (LP) T and B lymphocytes in inhibitory G-protein alpha2 subunit-deficient (Galphai2-/-) mice developing an IBD resembling ulcerative colitis was investigated following repeated oral immunizations with keyhole limpet haemocyanin (KLH), together with the adjuvant cholera toxin, prior to colitis. The antigen-specific B-cell response in the LP of both the small and the large intestines was significantly reduced in Galphai2-/- as compared to wild-type mice. In contrast, the frequency of KLH-specific immunoglobulin (Ig)-producing cells in the PP did not differ between Galphai2-/- and wild-type mice, whereas the total frequency of Ig-producing cells as well as the frequency of enteric flora-specific Ig-producing cells in the PP was significantly increased in Galphai2-/- as compared to wild-type mice. Analysis of T cell responses following restimulation ex vivo with KLH revealed a dramatic increase in the production of interferon-gamma in mesenteric lymph node, PP and LP lymphocytes from Galphai2 deficient as compared to wild-type mice, together with decreased production of interleukin-10 in all locations except the PP. PMID- 15885135 TI - Identification of T-cell epitopes on U1A protein in MRL/lpr mice: double-negative T cells are the major responsive cells. AB - Systemic lupus erythematosus (SLE) is characterized by the existence of a heterogeneous group of autoantibodies such as anti-DNA, chromatin, histone, and ribonucleoprotein antibodies (Abs). Although the B-cell antigenic determinants have been well characterized, very limited data about the T-cell epitopes of self antigen (Ag) have been reported. In the present study, we analysed auto-T-cell epitopes using bone marrow-derived dendritic cells (BM-DCs) pulsed with murine U1A (mU1A) protein capable of activating autoreactive T cells from unprimed MRL/lpr mice in vitro. The data suggested that there are at least four T-cell epitopes on the U1A protein, U1A31-50, U1A61-80, U1A201-220 and U1A271-287, and U1A31-50 had the most significant T-cell proliferative response. In addition, the main responsive T cells are the CD4- CD8- double-negative subgroup of T cells. Furthermore, we also demonstrated that the activation of double-negative T cells is major histocompatibility complex class II restricted. The study here provides information on T-cell epitope analysis of the U1A antigen using BM-DCs as the effective antigen-presenting cells. PMID- 15885136 TI - Morphological characterization of a human glioma cell l ine. AB - A human malignant continuous cell line, named NG97, was recently established in our laboratory. This cell line has been serially subcultured over 100 times in standard culture media presenting no sign of cell senescence. The NG97 cell line has a doubling time of about 24 h. Immunocytochemical analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP) and S-100 protein, and negative for vimentin. Under phase-contrast microscope, cultures of NG97 showed cells with variable morphological features, such as small rounded cells, fusiform cells (fibroblastic-like cells), and dendritic-like cells. However, at confluence just small rounded and fusiform cells can be observed. At scanning electron microscopy (SEM) small rounded cells showed heterogeneous microextentions, including blebs and filopodia. Dendritic-like cells were flat and presented extensive prolongations, making several contacts with small rounded cells, while fusiform cells presented their surfaces dominated by microvilli.We believe that the knowledge about NG97 cell line may be useful for a deeper understanding of biological and immunological characteristics of gliomas. PMID- 15885137 TI - Evolutionary explanations in medical and health profession courses: are you answering your students' "why" questions? AB - BACKGROUND: Medical and pre-professional health students ask questions about human health that can be answered in two ways, by giving proximate and evolutionary explanations. Proximate explanations, most common in textbooks and classes, describe the immediate scientifically known biological mechanisms of anatomical characteristics or physiological processes. These explanations are necessary but insufficient. They can be complemented with evolutionary explanations that describe the evolutionary processes and principles that have resulted in human biology we study today. The main goal of the science of Darwinian Medicine is to investigate human disease, disorders, and medical complications from an evolutionary perspective. DISCUSSION: This paper contrasts the differences between these two types of explanations by describing principles of natural selection that underlie medical questions. Thus, why is human birth complicated? Why does sickle cell anemia exist? Why do we show symptoms like fever, diarrhea, and coughing when we have infection? Why do we suffer from ubiquitous age-related diseases like arteriosclerosis, Alzheimer's and others? Why are chronic diseases like type II diabetes and obesity so prevalent in modern society? Why hasn't natural selection eliminated the genes that cause common genetic diseases like hemochromatosis, cystic fibrosis, Tay sachs, PKU and others? SUMMARY: In giving students evolutionary explanations professors should underscore principles of natural selection, since these can be generalized for the analysis of many medical questions. From a research perspective, natural selection seems central to leading hypotheses of obesity and type II diabetes and might very well explain the occurrence of certain common genetic diseases like cystic fibrosis, hemochromatosis, Tay sachs, Fragile X syndrome, G6PD and others because of their compensating advantages. Furthermore, armed with evolutionary explanations, health care professionals can bring practical benefits to patients by treating their symptoms of infection more specifically and judiciously. They might also help curtail the evolutionary arms race between pathogens and antibiotic defenses. PMID- 15885138 TI - Common polymorphism in H19 associated with birthweight and cord blood IGF-II levels in humans. AB - BACKGROUND: Common genetic variation at genes that are imprinted and exclusively maternally expressed could explain the apparent maternal-specific inheritance of low birthweight reported in large family pedigrees. We identified ten single nucleotide polymorphisms (SNPs) in H19, and we genotyped three of these SNPs in families from the contemporary ALSPAC UK birth cohort (1,696 children, 822 mothers and 661 fathers) in order to explore associations with size at birth and cord blood IGF-II levels. RESULTS: Both offspring's and mother's H19 2992C>T SNP genotypes showed associations with offspring birthweight (P = 0.03 to P = 0.003) and mother's genotype was also associated with cord blood IGF-II levels (P = 0.0003 to P = 0.0001). The offspring genotype association with birthweight was independent of mother's genotype (P = 0.01 to P = 0.007). However, mother's untransmitted H19 2992T allele was also associated with larger birthweight (P = 0.04) and higher cord blood IGF-II levels (P = 0.002), suggesting a direct effect of mother's genotype on placental IGF-II expression and fetal growth. The association between mother's untransmitted allele and cord blood IGF-II levels was more apparent in offspring of first pregnancies than subsequent pregnancies (P-interaction = 0.03). Study of the independent Cambridge birth cohort with available DNA in mothers (N = 646) provided additional support for mother's H19 2992 genotype associations with birthweight (P = 0.04) and with mother's glucose levels (P = 0.01) in first pregnancies. CONCLUSION: The common H19 2992T allele, in the mother or offspring or both, may confer reduced fetal growth restraint, as indicated by associations with larger offspring birth size, higher cord blood IGF II levels, and lower compensatory early postnatal catch-up weight gain, that are more evident among mother's smaller first-born infants. PMID- 15885139 TI - Monitoring of National Drug Policy (NDP) and its standardized indicators; conformity to decisions of the national drug selecting committee in Iran. AB - BACKGROUND: Pharmaceuticals have made an important contribution to global reductions in morbidity and mortality. To help save lives and improve health, it is important to be sure about equity to access to drugs, drug efficacy, quality and safety, and rational use of drugs, which are standardized National Drug Policy (NDP) objectives. NDP's indicators are useful to evaluate the pharmaceutical system performance in a country. Iran has adapted a National Drug List (NDL). Since management of drug supply in Iran takes place only for drugs that have been selected in NDL and this list is selected by the member of Iran Drug Selecting Committee (IDSC), thus evaluation of IDSC's decision making during last 5 years is an appropriate way to evaluate the implementation of drug supply system in the country. METHODS: To identify strengths and weaknesses of pharmaceutical policy formation and implementation in Iran, four standard questionnaires of the World Health Organization (WHO) were used. To assess the agreement between decisions of IDSC and standardized NDP indicators in the last 5 years (1998-2002), a weighted questionnaire by nominal group technique based on the questions that should be answered during discussion about one drug in IDSC was designed and used. RESULTS: There is a totally generics based NDP with 95% local production, that provides affordable access to drugs. The system, structures, and mechanisms were in place; however, they did not function properly in some topics. Assessment of 59 dossiers of approved drugs for adding to NDL during last 5 years showed that IDSC's members pay more attention to efficacy, safety, and rationality in use rather than accessibility and affordability. CONCLUSION: Revision of drug system in term of implementation of the processes to achieve NDP's objectives is necessary to save public health. Clarification of NDP's objectives and their impact for IDSC's members will result in improvement of the equity in access to pharmaceuticals. PMID- 15885140 TI - Liposome retention in size exclusion chromatography. AB - BACKGROUND: Size exclusion chromatography is the method of choice for separating free from liposome-encapsulated molecules. However, if the column is not presaturated with lipids this type of chromatography causes a significant loss of lipid material. To date, the mechanism of lipid retention is poorly understood. It has been speculated that lipid binds to the column material or the entire liposome is entrapped inside the void. RESULTS: Here we show that intact liposomes and their contents are retained in the exclusion gel. Retention depends on the pore size, the smaller the pores, the higher the retention. Retained liposomes are not tightly fixed to the beads and are slowly released from the gels upon direct or inverted eluent flow, long washing steps or column repacking. Further addition of free liposomes leads to the elution of part of the gel trapped liposomes, showing that the retention is transitory. Trapping reversibility should be related to a mechanism of partitioning of the liposomes between the stationary phase, water-swelled polymeric gel, and the mobile aqueous phase. CONCLUSION: Retention of liposomes by size exclusion gels is a dynamic and reversible process, which should be accounted for to control lipid loss and sample contamination during chromatography. PMID- 15885141 TI - Play dough as an educational tool for visualization of complicated cerebral aneurysm anatomy. AB - BACKGROUND: Imagination of the three-dimensional (3D) structure of cerebral vascular lesions using two-dimensional (2D) angiograms is one of the skills that neurosurgical residents should achieve during their training. Although ongoing progress in computer software and digital imaging systems has facilitated viewing and interpretation of cerebral angiograms enormously, these facilities are not always available. METHODS: We have presented the use of play dough as an adjunct to the teaching armamentarium for training in visualization of cerebral aneurysms in some cases. RESULTS: The advantages of play dough are low cost, availability and simplicity of use, being more efficient and realistic in training the less experienced resident in comparison with the simple drawings and even angiographic views from different angles without the need for computers and similar equipment. The disadvantages include the psychological resistance of residents to the use of something in surgical training that usually is considered to be a toy, and not being as clean as drawings or computerized images. CONCLUSION: Although technology and computerized software using the patients' own imaging data seems likely to become more advanced in the future, use of play dough in some complicated cerebral aneurysm cases may be helpful in 3D reconstruction of the real situation. PMID- 15885142 TI - Trends in provision of photodynamic therapy and clinician attitudes: a tracker survey of a new health technology. AB - BACKGROUND: There has been debate about the cost-effectiveness of photodynamic therapy (PDT), a treatment for neovascular age-related macular degeneration. We have been monitoring trends for the provision of PDT in the UK National Health Service. The fourth annual 'tracker' survey took place as definitive National Institute for Clinical Excellence (NICE) guidance was issued. We assessed trends in PDT provision up to the point of release of the NICE guidance and identified likely sources of pressure on ophthalmologists to provide PDT. METHODS: National postal questionnaire survey of clinicians with potential responsibility for PDT provision. The survey explored reported local provision, beliefs about the effectiveness of PDT and what sources of opinion might influence attitudes towards providing PDT. RESULTS: The response rate was 73% (111/150). Almost half of the surveyed ophthalmology units routinely provided PDT, as part of a trend of steady growth in provision. The proportion of respondents who believed that further proof of effectiveness was required has also declined despite the absence of any new substantial evidence. Attitudes towards providing PDT were positive, on average, and were more strongly associated with perceived social pressure from local colleagues than from other sources. Local colleagues were seen as being most approving of PDT. CONCLUSION: Those responsible for implementing the NICE guidance need to address ophthalmologists' beliefs about the evidence of effectiveness for PDT and draw upon supportive local individuals or networks to enhance the credibility of the guidance. PMID- 15885143 TI - The costs and effects of a nationwide insecticide-treated net programme: the case of Malawi. AB - BACKGROUND: Insecticide-treated nets (ITNs) are a proven intervention to reduce the burden of malaria, yet there remains a debate as to the best method of ensuring they are universally utilized. This study is a cost-effectiveness analysis of an intervention in Malawi that started in 1998, in Blantyre district, before expanding nationwide. Over the 5-year period, 1.5 million ITNs were sold. METHODS: The costs were calculated retrospectively through analysis of expenditure data. Costs and effects were measured as cost per treated-net year (cost/TNY) and cost per net distributed. RESULTS: The mean cost/TNY was calculated at $4.41, and the mean cost/ITN distributed at $2.63. It also shows evidence of economies of scale, with the cost/TNY falling from $7.69 in year one (72,196 ITN) to $3.44 in year five (720,577 ITN). Cost/ITN distributed dropped from $5.04 to $1.92. CONCLUSION: Combining targeting and social marketing has the potential of being both cost-effective and capable of achieving high levels of coverage, and it is possible that increasing returns to scale can be achieved. PMID- 15885144 TI - A simple technique for quantifying apoptosis in 96-well plates. AB - BACKGROUND: Analyzing apoptosis has been an integral component of many biological studies. However, currently available methods for quantifying apoptosis have various limitations including multiple, sometimes cell-damaging steps, the inability to quantify live, necrotic and apoptotic cells at the same time, and non-specific detection (i.e. "false positive"). To overcome the shortcomings of current methods that quantify apoptosis in vitro and to take advantage of the 96 well plate format, we present here a modified ethidium bromide and acridine orange (EB/AO) staining assay, which may be performed entirely in a 96-well plate. Our method combines the advantages of the 96-well format and the conventional EB/AO method for apoptotic quantification. RESULTS: We compared our method and the conventional EB/AO method for quantifying apoptosis of suspension cells (Jurkat) and adherent cells (A375) under normal growth and apoptosis inducing conditions. We found that our new EB/AO method achieved quantification results comparable to those produced using the conventional EB/AO method for both suspension and adherent cells. CONCLUSION: By eliminating the detaching and washing steps, our method drastically reduces the time needed to perform the test, minimizes damage to adherent cells, and decreases the possibility of losing floating cells. Overall, our method is an improvement over the currently available techniques especially for adherent cells. PMID- 15885145 TI - Integration of the Gene Ontology into an object-oriented architecture. AB - BACKGROUND: To standardize gene product descriptions, a formal vocabulary defined as the Gene Ontology (GO) has been developed. GO terms have been categorized into biological processes, molecular functions, and cellular components. However, there is no single representation that integrates all the terms into one cohesive model. Furthermore, GO definitions have little information explaining the underlying architecture that forms these terms, such as the dynamic and static events occurring in a process. In contrast, object-oriented models have been developed to show dynamic and static events. A portion of the TGF-beta signaling pathway, which is involved in numerous cellular events including cancer, differentiation and development, was used to demonstrate the feasibility of integrating the Gene Ontology into an object-oriented model. RESULTS: Using object-oriented models we have captured the static and dynamic events that occur during a representative GO process, "transforming growth factor-beta (TGF-beta) receptor complex assembly" (GO:0007181). CONCLUSION: We demonstrate that the utility of GO terms can be enhanced by object-oriented technology, and that the GO terms can be integrated into an object-oriented model by serving as a basis for the generation of object functions and attributes. PMID- 15885146 TI - Pigs in sequence space: a 0.66X coverage pig genome survey based on shotgun sequencing. AB - BACKGROUND: Comparative whole genome analysis of Mammalia can benefit from the addition of more species. The pig is an obvious choice due to its economic and medical importance as well as its evolutionary position in the artiodactyls. RESULTS: We have generated approximately 3.84 million shotgun sequences (0.66X coverage) from the pig genome. The data are hereby released (NCBI Trace repository with center name "SDJVP", and project name "Sino-Danish Pig Genome Project") together with an initial evolutionary analysis. The non-repetitive fraction of the sequences was aligned to the UCSC human-mouse alignment and the resulting three-species alignments were annotated using the human genome annotation. Ultra-conserved elements and miRNAs were identified. The results show that for each of these types of orthologous data, pig is much closer to human than mouse is. Purifying selection has been more efficient in pig compared to human, but not as efficient as in mouse, and pig seems to have an isochore structure most similar to the structure in human. CONCLUSION: The addition of the pig to the set of species sequenced at low coverage adds to the understanding of selective pressures that have acted on the human genome by bisecting the evolutionary branch between human and mouse with the mouse branch being approximately 3 times as long as the human branch. Additionally, the joint alignment of the shot-gun sequences to the human-mouse alignment offers the investigator a rapid way to defining specific regions for analysis and resequencing. PMID- 15885147 TI - Using the ecology model to describe the impact of asthma on patterns of health care. AB - BACKGROUND: Asthma changes both the volume and patterns of healthcare of affected people. Most studies of asthma health care utilization have been done in selected insured populations or in a single site such as the emergency department. Asthma is an ambulatory sensitive care condition making it important to understand the relationship between care in all sites across the health service spectrum. Asthma is also more common in people with fewer economic resources making it important to include people across all types of insurance and no insurance categories. The ecology of medical care model may provide a useful framework to describe the use of health services in people with asthma compared to those without asthma and identify subgroups with apparent gaps in care. METHODS: This is a case-control study using the 1999 U.S. Medical Expenditure Panel Survey. Cases are school-aged children (6 to 17 years) and young adults (18 to 44 years) with self-reported asthma. Controls are from the same age groups who have no self-reported asthma. Descriptive analyses and risk ratios are placed within the ecology of medical care model and used to describe and compare the healthcare contact of cases and controls across multiple settings. RESULTS: In 1999, the presence of asthma significantly increased the likelihood of an ambulatory care visit by 20 to 30% and more than doubled the likelihood of making one or more visits to the emergency department (ED). Yet, 18.8% of children and 14.5% of adults with asthma (over a million Americans) had no ambulatory care visits for asthma. About one in 20 to 35 people with asthma (5.2% of children and 3.6% of adults) were seen in the ED or hospital but had no prior or follow-up ambulatory care visits. These Americans were more likely to be uninsured, have no usual source of care and live in metropolitan areas. CONCLUSION: The ecology model confirmed that having asthma changes the likelihood and pattern of care for Americans. More importantly, the ecology model identified a subgroup with asthma who sought only emergent or hospital services. PMID- 15885148 TI - Cost-effectiveness of recommended nurse staffing levels for short-stay skilled nursing facility patients. AB - BACKGROUND: Among patients in skilled nursing facilities for post-acute care, increased registered nurse, total licensed staff, and nurse assistant staffing is associated with a decreased rate of hospital transfer for selected diagnoses. However, the cost-effectiveness of increasing staffing to recommended levels is unknown. METHODS: Using a Markov cohort simulation, we estimated the incremental cost-effectiveness of recommended staffing versus median staffing in patients admitted to skilled nursing facilities for post-acute care. The outcomes of interest were life expectancy, quality-adjusted life expectancy, and incremental cost-effectiveness. RESULTS: The incremental cost-effectiveness of recommended staffing versus median staffing was $321,000 per discounted quality-adjusted life year gained. One-way sensitivity analyses demonstrated that the cost effectiveness ratio was most sensitive to the likelihood of acute hospitalization from the nursing home. The cost-effectiveness ratio was also sensitive to the rapidity with which patients in the recommended staffing scenario recovered health-related quality of life as compared to the median staffing scenario. The cost-effectiveness ratio was not sensitive to other parameters. CONCLUSION: Adopting recommended nurse staffing for short-stay nursing home patients cannot be justified on the basis of decreased hospital transfer rates alone, except in facilities with high baseline hospital transfer rates. Increasing nurse staffing would be justified if health-related quality of life of nursing home patients improved substantially from greater nurse and nurse assistant presence. PMID- 15885149 TI - Using autoregressive integrated moving average (ARIMA) models to predict and monitor the number of beds occupied during a SARS outbreak in a tertiary hospital in Singapore. AB - BACKGROUND: The main objective of this study is to apply autoregressive integrated moving average (ARIMA) models to make real-time predictions on the number of beds occupied in Tan Tock Seng Hospital, during the recent SARS outbreak. METHODS: This is a retrospective study design. Hospital admission and occupancy data for isolation beds was collected from Tan Tock Seng hospital for the period 14th March 2003 to 31st May 2003. The main outcome measure was daily number of isolation beds occupied by SARS patients. Among the covariates considered were daily number of people screened, daily number of people admitted (including observation, suspect and probable cases) and days from the most recent significant event discovery. We utilized the following strategy for the analysis. Firstly, we split the outbreak data into two. Data from 14th March to 21st April 2003 was used for model development. We used structural ARIMA models in an attempt to model the number of beds occupied. Estimation is via the maximum likelihood method using the Kalman filter. For the ARIMA model parameters, we considered the simplest parsimonious lowest order model. RESULTS: We found that the ARIMA (1,0,3) model was able to describe and predict the number of beds occupied during the SARS outbreak well. The mean absolute percentage error (MAPE) for the training set and validation set were 5.7% and 8.6% respectively, which we found was reasonable for use in the hospital setting. Furthermore, the model also provided three-day forecasts of the number of beds required. Total number of admissions and probable cases admitted on the previous day were also found to be independent prognostic factors of bed occupancy. CONCLUSION: ARIMA models provide useful tools for administrators and clinicians in planning for real-time bed capacity during an outbreak of an infectious disease such as SARS. The model could well be used in planning for bed-capacity during outbreaks of other infectious diseases as well. PMID- 15885150 TI - Characterisation and application of a bovine U6 promoter for expression of short hairpin RNAs. AB - BACKGROUND: The use of small interfering RNA (siRNA) molecules in animals to achieve double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful method of sequence-specific gene knockdown. As DNA-based expression of short hairpin RNA (shRNA) for RNAi may offer some advantages over chemical and in vitro synthesised siRNA, a number of vectors for expression of shRNA have been developed. These often feature polymerase III (pol. III) promoters of either mouse or human origin. RESULTS: To develop a shRNA expression vector specifically for bovine RNAi applications, we identified and characterised a novel bovine U6 small nuclear RNA (snRNA) promoter from bovine sequence data. This promoter is the putative bovine homologue of the human U6-8 snRNA promoter, and features a number of functional sequence elements that are characteristic of these types of pol. III promoters. A PCR based cloning strategy was used to incorporate this promoter sequence into plasmid vectors along with shRNA sequences for RNAi. The promoter was then used to express shRNAs, which resulted in the efficient knockdown of an exogenous reporter gene and an endogenous bovine gene. CONCLUSION: We have mined data from the bovine genome sequencing project to identify a functional bovine U6 promoter and used the promoter sequence to construct a shRNA expression vector. The use of this native bovine promoter in shRNA expression is an important component of our future development of RNAi therapeutic and transgenic applications in bovine species. PMID- 15885159 TI - [Succession and innovation of Chinese traditional surgery: a perspective on the history]. AB - Facing the challenge of modern medicine, how to develop Chinese traditional surgery is a question. The tremendous heritage of traditional Chinese medicine (TCM) is the springhead of the development and innovation of TCM. Succession and innovation is an eternal subject for the development of Chinese traditional surgery. In order to improve Chinese traditional surgery, we should explore traditional medical literature, and carry forward the theories of Chinese traditional surgery. PMID- 15885160 TI - [Challenges and opportunities for the development of Chinese traditional surgery]. AB - Chinese traditional surgery, an important branch of traditional Chinese medicine, has contributed greatly to the people's health in the history. With the fast development of modern medicine, Chinese traditional surgery is faced with serious challenges and its shortcomings have appeared more and more evident. In this article, the authors analyzed some problems in the development of Chinese traditional surgery and the opportunities for its further development, and suggested that the superiorities of Chinese traditional surgery should be enhanced in its future development. PMID- 15885161 TI - [Discussion of some problems about use of traditional Chinese medicine in perioperative management of breast cancer]. AB - The study on use of traditional Chinese medicine in perioperative management of breast cancer is still in the beginning phase. With the emergence of new understanding about the biological characteristics of breast cancer, the concept of treatment has changed. For instance, the resection extent is tending to be narrower, large doses of radiotherapy may be adopted during the operation, and early use of adjuvant chemotherapy is advocated after the operation. These have bought about changes to the intervention factors in the perioperative period for breast cancer. Some related problems about the participation of traditional Chinese medicine in perioperative management of breast cancer are discussed in this article, so as to make the perioperative management perfect. PMID- 15885162 TI - [Pondering the problems of clinical effect assessment of traditional Chinese medicine]. AB - Clinical effect is of great importance to traditional Chinese medicine (TCM), and the rigorous and scientific methods for clinical effect assessment should be highly stressed in the clinical research of TCM. In this article, the authors analyzed the current status of the effect assessment of TCM and proposed some measures to improve the current effect assessment of TCM, in the hope of establishing an internationally accepted system of effect assessment reflecting the superiority and characteristics of TCM therapies. PMID- 15885163 TI - [General methods for clinical effect assessment of traditional Chinese medicine]. AB - Along with the changes of disease spectrum, medical models and health concept, people pay more and more attention to the diagnostic and therapeutic approaches of traditional Chinese medicine (TCM). With the effect assessment being changed from disease-based model to patient-based model, how to scientifically and objectively explain the validity of TCM has become the premise for further development of TCM and dissemination of it throughout the world. In this article, the authors analyzed the status quo and problems of the effect assessment of TCM, and proposed some general methods for clinical effect assessment of TCM, including formulating criteria for syndrome differentiation under the guidance of TCM theories, paying attention to quality of life, proper selection of indexes for outcome assessment, application of modern clinical study methods, such as the methods of evidence-based medicine and clinical epidemiology, designing randomized controlled trials, multi-subject co-operation, strict supervision of the quality of researches, and establishment of organizations for professional training. PMID- 15885164 TI - [Prospective randomized trial of optimum anal canal release treatment for chronic anal fissure]. PMID- 15885165 TI - [Discussion on design and quality control of clinical trials of traditional Chinese medicine on the basis of Fuhuang Tablets clinical study]. AB - OBJECTIVES: To assess the efficacy of Fuhuang Tablets in treating internal hemorrhoid hemorrhage and discussing the design and quality control of clinical trials of traditional Chinese medicine. METHODS: According to two-center, randomized, blinded and controlled study, 144 patients with internal hemorrhoids were divided into 2 groups. Seventy-two patients were treated with Fuhuang Tablets (4 tablets t.i.d. per os) and the other 72 patients were treated with Zhining Tablets (4 tablets t.i.d. per os). Symptoms and signs of the patients were recorded and evaluated at the third and seventh day of treatment respectively. RESULTS: At the third day of treatment, the overall response rates of Fuhuang Tablets-treated group and Zhining Tablets-treated group were 58.57% and 27.78% respectively, with statistic difference between the two groups (P<0.05). At the seventh day of treatment, the overall response rates of Fuhuang Tablets-treated group and Zhining Tablets-treated group were 91.43% and 81.94% respectively, with no statistic difference between the two groups. The laboratory test results and life signs of the 144 patients were normal before and after the treatment. CONCLUSION: Fuhuang Tablets are effective and safe in treating internal hemorrhoid hemorrhage. PMID- 15885166 TI - [Treatment of refractory carbuncle of the nape with Herba Taraxaci: a report of one case]. PMID- 15885167 TI - [Effects of perioperative administration of Rhubarb on acute inflammatory response in patients with gastric cancer]. AB - OBJECTIVE: To observe the effect of perioperative administration of rhubarb on the acute inflammatory response in patients with gastric cancer. METHODS: In this prospective, single-blinded, controlled clinical trial, thirty-one patients with gastric cancer operatively treated were randomly divided into two groups, with 14 patients in control group and 17 in study group. Patients in both groups were given an isocaloric and isonitrogenous enteral diet. The enteral diet was started 36 hours after operation, and continued for 6 days. Patients in the study group were fed with rhubarb before operation, and at 1 day and 2 days after operation. Indexes of acute inflammatory response such as serum C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), and indexes of nutritional status such as serum albumin (ALB), prealbumin (PA) and transferrin (TRF) were measured before operation, and at 1 day, 3 and 7 days after operation. RESULTS: Patients in both groups had acute inflammatory response, and the indexes of nutritional status decreased after operation.IL-6, CRP and TNF-alpha tested at 3 and 7 days after operation were lower in the study group as compared with those in the control group, and the recovery time of gastrointestinal motility such as borborygmus, gas elimination and defecation was shorter in the study group as compared with that in the control group. The indexes of nutritional status showed no significant differences between two groups after operation. CONCLUSION: Rhubarb can positively modulate the acute inflammatory response, promote the recovery of postoperative gastrointestinal motility, and benefit enteral nutrition support in patients who have undergone major operations for gastric cancer. PMID- 15885168 TI - [Diagnosis of plasma cell mastitis with multi-slice spiral CT]. AB - OBJECTIVE: To evaluate the diagnostic value of multi-slice spiral CT (MSCT) for plasma cell mastitis. METHODS: Radiographs of MSCT for forty-six patients with plasma cell mastitis diagnosed by pathological examination were reviewed. RESULTS: The findings of MSCT of plasma cell mastitis could be divided into four types, including the inflammation type, the abscess type, the sinus and fistula type, and the mixed type, and each type had its radiographic characteristics. CONCLUSION: MSCT is helpful for diagnosing plasma cell mastitis and should be used as an examination of first choice for the patients. PMID- 15885169 TI - [Color Doppler ultrasound study on pathological changes of lower extremity arteries in patients with metabolic syndrome]. AB - OBJECTIVE: To explore the pathological changes of lower extremity arteries of patients with metabolic syndrome. METHODS: Fifty patients with metabolic syndrome (MS) and 30 normal volunteers were included in this study. The inner diameters, peak systolic velocities and intima-media thicknesses of the lower extremity arteries, and the sizes and numbers of the plaques in the lower extremity arteries were obtained with color Doppler ultrasound (CDU). RESULTS: In the fifty MS patients, the intima-media thicknesses of forty-one were thicker than 1.1 mm and thirty-three had plaques which had brought stenosis in different degrees. The intima-media thicknesses of the common femoral, popliteal and posterior tibial arteries in the MS patients were significantly thicker than those in the normal volunteers (P<0.01 or P<0.05), and a greater number of plaques in the lower extremity arteries were observed in the patients, as compared with the normal volunteers (P<0.01). CONCLUSION: Various pathological changes had taken place in the lower extremity arteries of the MS patients. The sites and degrees of the lower extremity arterial diseases in MS patients can be definitely detected with CDU. PMID- 15885170 TI - [Effects of Shengqing Capsules on cholelithiasis-related genes in guinea pigs]. AB - OBJECTIVE: To explore the molecular mechanisms of Shengqing Capsules in treating cholelithiasis. METHODS: Sixty female guinea pigs were randomized into 3 groups: group I (fed with normal diet), group II (fed with low-protein diet) and group III (fed with low-protein diet and Shengqing Capsules). After six-week feeding, the gallstone formation and the expressions of bilirubin UDP glucuronosyltransferase (B-UGT) mRNA and cholesterol 7alpha-hydroxylase (CYP7A1) mRNA were observed. RESULTS: The proportions of stone-formed in groups I, II and III were 2/14, 9/12 and 4/14, respectively. There were significant differences among the three groups (P<0.05). The expressions of B-UGT and CYP7A1 mRNAs were higher in both group I and group III as compared with those in the group II (P<0.05). CONCLUSION: Shengqing Capsules can reduce the rate of stone-formation, which may be due to its interference of metabolism of bilirubin and cholesterol and up-regulation of the expressions of B-UGT and CYP7A1 mRNAs. PMID- 15885171 TI - [Impact of antipyretic and purgative herbs on intestinal mucosal barrier and inflammatory response in treatment of acute cholangitis in rats]. AB - OBJECTIVE: To investigate the effects of antipyretic and purgative herbs on intestinal mucosal barrier and inflammatory response in the treatment of acute cholangitis. METHODS: Sixty SD rats were randomly divided into group A (untreated group, acute cholangitis was induced, n=20), group B (treatment group, acute cholangitis was induced and treated with antipyretic and purgative herbs, n=20) and group C (sham operation group, n=20). At the third or fifth day after operation, the rats were sacrificed and sampled. The serum endotoxin, cytokines and inflammatory mediators were tested and the numbers of labeled bacteria in the liver, spleen and mesenteric lymph nodes translocated from the gut were assayed. RESULTS: As compared with group A, the serum content of endotoxin, IL-6, IL-8, TNF-alpha, CRP and NO was significantly lower and that of IL-2 was significantly higher, and the translocated numbers of labeled bacteria from gut were reduced in both group B and group C (P<0.01). CONCLUSION: Antipyretic and purgative herbs can play therapeutic roles in the treatment of acute biliary tract infections, including the protection of intestinal mucosal barrier from bacterial translocation, reduction of serum endotoxin content and regulation of inflammatory response. PMID- 15885172 TI - [Effects of Shengji Huayu Recipe and its decomposed formulas on synthesis of collagen types I and III in granulation tissue of rats in early wound healing]. AB - OBJECTIVE: To study the effects of Shengji Huayu Recipe (a traditional Chinese medicine compound recipe for resolving stagnation and promoting granulation) and its decomposed formulas (Huayu Recipe for resolving stagnation and Shengji Recipe for promoting granulation) on the synthesis of collagen types I and III in granulation tissue of rats in early wound healing. METHODS: Twenty-four male Sprague-Dawley (SD) rats with full-thickness skin lesion were randomized into 4 groups: Shengji Huayu Recipe-treated group, Shengji Recipe-treated group, Huayu Recipe-treated group and untreated group. Collagen types I and III in granulation tissue of the rats were tested with immunohistochemical methods and image analysis. RESULTS: On the third day of wound healing, collagen I of the rats in both Shengji Huayu Recipe-treated group and Shengji Recipe-treated group was higher than that in the untreated group, and collagen I of the rats in Huayu Recipe-treated group was lower than that in the untreated group (P<0.05). Collagen III of the rats in the three treated groups were lower than that in the untreated group (P<0.05). On the seventh day of wound healing, Collagen I of the rats in both Shengji Huayu Recipe-treated group and Shengji Recipe-treated group was higher than that in the untreated group (P<0.05), and collagen III of the rats in both Shengji Recipe-treated group and Huayu Recipe-treated group was higher than that in the untreated group (P<0.05). CONCLUSION: Resolving stagnation and promoting granulation therapy can promote the wound healing in rats. PMID- 15885173 TI - [Effects of Wenshen Jianpi Recipe on chronic wound healing in rats]. AB - OBJECTIVE: To study the effects of Wenshen Jianpi Recipe (WSJPR, a traditional Chinese medicine for warming kidney and invigorating spleen) on chronic wound healing and the mechanism. METHODS: Ninety-six SD rats were randomly divided into 4 groups, with 24 rats in each group, and back wound was made in the rats. For rats in 3 of the 4 groups, hydrocortisone injection was administered to induce chronic wound. Rats in 2 of the 3 groups were treated with WSJPR and Xinpukang Granules (XPKG) respectively, and the rats in the other group were untreated. The rats in the fourth group were taken as control. The wound healing time and the width of new epidermis were observed, and the histomorphological changes and cell cycle of the granulation tissue, and the protein expressions of epidermal growth factor (EGF), transforming growth factor-beta(1) (TGF-beta(1)) and fibronectin (FN) in the granulation tissue were tested with immunohistochemical technique and flow cytometry. RESULTS: The wound healing time of the WSJPR-treated and XPKG treated groups was (17.0+/-1.9) and (18.8+/-1.9) d respectively, much shorter than that of the untreated and control groups (P<0.05). On the 14th experiment day, the width of new epidermis of the WSJPR-treated and XPKG-treated groups was (3.73+/-0.19) and (3.21+/-0.15) mm respectively, much wider than that of the untreated and control groups (P<0.05). The numbers of angiogenesis, fibroblasts and cells in the S phase in WSJPR-treated and XPKG-treated groups were much higher than those in the untreated and control groups (P<0.05). Compared with the untreated and control groups, the protein expressions of EGF, TGF-beta(1) and FN in WSJPR-treated and XPKG-treated groups were higher (P<0.05). CONCLUSION: WSJPR can enhance the wound healing. It was likely through accelerating the cell proliferation and up-regulating the expressions of EGF, TGF-beta(1) and FN. PMID- 15885174 TI - [Effects of Shujin Zhuanggu Tablets on fracture healing of metacarpus: a report of 225 cases]. PMID- 15885175 TI - [Effects of Runing Recipe II on expressions of p53 and ras oncogene proteins and cell cycle of the transplanted Ca761 breast cancer in mice]. AB - OBJECTIVE: To explore the mechanisms of Runing Recipe II (a recipe composed of traditional Chinese herbs) in inhibiting the growth of breast cancer by observing its effects on the expressions of p53 and ras oncogene proteins and cell cycle of the transplanted Ca761 breast cancer in mice. METHODS: We established the breast cancer model by transplanting Ca761 cells in mice. The mice were randomly divided into 4 groups: normal saline control group, CTX-treated group, Runing Recipe II treated group, and Runing Recipe II and CTX-treated group, with 12 mice in each group. We detected the cell cycle of the cancer cells in the mice's transplanted tumor with flow cytometry and measured the expressions of p53 and ras oncogene proteins in the transplanted tumor with immunohistochemical methods. RESULTS: The percentages of tumor cells in S-phase of the Runing Recipe II treated group, CTX treated group and Runing Recipe II and CTX-treated group were significantly lower than that of the normal saline control group respectively (P<0.05). The percentage of tumor cells in G(0)-G(1) phase of the Runing Recipe II treated group was lower than that of the CTX-treated group (P<0.05), while the percentage of tumor cells in G(2)-M phase was higher than that of the CTX-treated group. The immunoreactive scores (IRSs) of p53 in the Runing Recipe II treated group and Runing Recipe II and CTX-treated group were significantly lower than that in the normal saline control group respectively (P<0.05). The effect of CTX on the expression of p53 was not significant. The IRSs of ras oncogene protein in the Recipe II-treated group, CTX-treated group and Runing Recipe II and CTX-treated group were lower than that in the normal saline control group respectively (P<0.05). CONCLUSION: Runing Recipe II can inhibit the growth of Ca761 breast cancer in mice by controlling the cell cycle of the transplanted tumor. This may be related to its effect on the gene expressions of p53 and ras in the tumor tissue. PMID- 15885176 TI - [Treatment of complex high anal fistula with combined use of anatomical fistulectomy and thread-drawing therapy: a report of 36 cases]. PMID- 15885177 TI - [Treatment of simple low anal fistula of infant with thread-drawing therapy: a report of 21 cases]. PMID- 15885178 TI - [Treatment of acute perianal necrotizing fasciitis with integrated traditional Chinese and Western medicine: a report of 9 cases]. PMID- 15885179 TI - [Professor Tang Han-Jun's experience in treating postoperative fistulas]. PMID- 15885180 TI - [Professor Tang Han-Jun' experience in treating unhealed wound after mastectomy for breast cancer]. PMID- 15885181 TI - [Research strategy and clinical significance of anorectum manometry]. PMID- 15885182 TI - [Study on wound healing mechanism of replenishing qi and dissipating stagnation therapy for chronic skin ulcers]. PMID- 15885183 TI - [On truthfulness in light of the English translation of the nomenclature of traditional Chinese medicine]. PMID- 15885184 TI - [Words expressing category in Chinese-English translation of traditional Chinese medicine]. PMID- 15885185 TI - [Telomeres, diseases and aging]. PMID- 15885186 TI - [Drugs of abuse, protein phosphatases, and ERK pathway]. PMID- 15885187 TI - [What do we learn from the genome of Legionella pneumophila?]. PMID- 15885188 TI - [Netherton syndrome: a model for studying the regulation of the desquamation process]. PMID- 15885189 TI - [From cadherins to the quantification of cell adhesion]. PMID- 15885190 TI - [Renal vascular and glomerular fibrosis and epidermal growth factor receptor transactivation]. PMID- 15885191 TI - [Plasmodium chloroquine resistance: towards a specific attack?]. PMID- 15885192 TI - [A new model for intra-embryonic hematopoietic stem cell generation]. PMID- 15885193 TI - [14-3-3 gamma, a novel partner of MuSK downregulates synaptic gene transcription at the neuromuscular junction]. PMID- 15885194 TI - [Influence of climate upon the meningitis onset in West Africa]. PMID- 15885195 TI - [Stimulation of proteoglycan synthesis towards cartilage repair]. PMID- 15885197 TI - [Neural differentiation of murine embryonic stem cells ES]. AB - Pluripotent murine embryonic stem (ES) cells can differentiate into all cell types both in vivo and in vitro. Based on their capability to proliferate and differentiate, these ES cells appear as a very promising tool for cell therapy. The understanding of the molecular mechanisms underlying the neural differentiation of the ES cells is a pre-requisite for selecting adequately the cells and conditions which will be able to correctly repair damaged brain and restore altered cognitive functions. Different methods allow obtaining neural cells from ES cells. Most of the techniques differentiate ES cells by treating embryoid bodies in order to keep an embryonic organization. More recent techniques, based on conditioned media, induce a direct differentiation of ES cells into neural cells, without going through the step of embryonic bodies. Beyond the fact that these techniques allow obtaining large numbers of neural precursors and more differentiated neural cells, these approaches also provide valuable information on the process of differentiation of ES cells into neural cells. Indeed, sequential studies of this process of differentiation have revealed that globally ES cells differentiating into neural cells in vitro recapitulate the molecular events governing the in vivo differentiation of neural cells. Altogether these data suggest that murine ES cells remain a highly valuable tool to obtain large amounts of precursor and differentiated neural cells as well as to get a better understanding of the mechanisms of neural differentiation, prior to a potential move towards the use of human ES cells in therapy. PMID- 15885198 TI - [Senescence: a telomeric limit to immortality or a cellular response to physiologic stresses?]. AB - Cells entering a state of senescence undergo a irreversible cell cycle arrest, associated by a set of functional and morphological changes. Senescence occurs following telomeres shortening (replicative senescence) or exposure to other acute or chronic physiologic stress signals (a phenomenon termed stasis: stress or aberrant signaling-induced senescence). In this review, I discuss the pathways of cellular senescence, the mechanisms involved and the role that these pathways have in regulating the initiation and progression of cancer. Telomere-initiated senescence or loss of telomere function trigger focal recruitement of protein sensors of the DNA double-strand breaks leading to the activation of the DNA damage checkpoint responses and the tumour suppressor gene product, p53, which in turn induces the cell-cycle inhibitor, p21(WAF1). Loss of p53 and pRb function allows continued cell division despite increasing telomere dysfunction and eventually entry into telomere crisis. Immortalisation is an essential prerequisite for the formation of a tumour cell. Therefore, a developing tumour cell must circumvent at least two proliferative barriers--cellular senescence and crisis--to achieve neoplastic transformation. These barriers are regulated by telomere shortening and by the p16(INK4a)/Rb and p53 tumour suppressor pathways. Elucidation of the genes and emerging knowledge about the regulatory mechanisms that lead to senescence and determine the pattern of gene expression in senescent cells may lead to more effective treatments for cancer. PMID- 15885199 TI - [Matrilysin-1 and cancer pathology]. AB - Matrilysin-1 has a considerable importance in the progression of cancers, namely digestive, ones inasmuch as it fulfils a direct function of deterioration on molecular targets at different levels and interferes as well in the activation of other proteolytic systems. Its low expression in sound tissues increases distinctly in mild cellular proliferations like adenomas to increase even more in carcinomas and be involved at the level of tumoral development as well as in a metastatical proliferation. It acts during the different stages of malignant transformation, splitting the E-cadherin, an intercellular adhesive molecule of epithelia, matrix molecules in the tumor-stroma interface and integrins which link matrix components to epithelial cells. The use of oligonucleotide antisens to the matrilysin-1 drives to a clear reduction of invading process, suggesting therefore an important therapeutic potential. PMID- 15885200 TI - [Happy birthday, CD34!]. AB - Twenty years after the initial description of the CD34 antigen, scientists are still interested in elucidating the exact function of a prototypic << stem cell antigen >> in human, primate and mouse models. While this question remains largely unresolved, this has not precluded the development of medical tools using the detection of CD34 in several applications that range from diagnosis of malignant blood diseases through the biological monitoring of haematopoietic cell collection for autologous or allogeneic transplantation. This review focuses on major aspects of CD34 biology. PMID- 15885201 TI - [A role of PPARgamma in reproduction?]. AB - Synthetic molecules of the glitazone family are currently used in the treatment of type II diabetes. Glitazones also improve secondary pathologies that are frequently associated with insulin resistance such as the polycystic ovary syndrome (PCOS). Glitazones bind to the peroxysome proliferator-activated receptor gamma (PPARgamma), a nuclear receptor which is highly expressed in adipose tissue. PPARgamma also binds natural ligands such as long-chain fatty acids. Recently, several groups have shown that PPARgamma is also highly expressed in ovarian granulosa cells, and that glitazones are able to modulate in vitro granulosa cell proliferation and steroidogenesis in several species. These recent data raise new questions concerning the underlying mechanism of the effect of glitazones on PCOS. One might hypothesize, as for other << glucophage >> molecules such as metformin, that it is the general improvement of glucose metabolism and insulin sensitivity by glitazones which indirectly, and via an unknown mechanism, ameliorates ovarian functionality. The data discussed here suggest now an alternative possibility, that glitazones act directly at the ovarian level. Moreover, PPARgamma also seems to play a key role in the maturation of the placenta. In particular, inactivation of PPARgamma in mice is lethal, since the foetus is unable to develop because of alterations of placental maturation. In women, the activation of PPARgamma in placenta leads to an increase of placental hormone secretion. Overall, these results raise some questions about the role of natural ligands of PPARgamma such as long chain fatty acids on female fertility and the interactions between energy metabolism and reproduction in general. PMID- 15885202 TI - [Inherited metabolic diseases: benefits of metabolomics]. AB - Both hope and illusion, the recent progress in biology has raised our expectations that one day it will be possible to introduce a biological sample into an apparatus which will then deliver in a few minutes thousands of qualitative and quantitative data concerning the genome, transcriptome, proteome and metabolome, thereby contributing to diagnosis and follow-up of diseases which are now difficult to identify. Such machines do not exist yet and, in any case, should be associated with the appropriate and adequate clinical work on the disease and with the patient. This << total >> approach is of course being pushed by the recent decoding of the genomes of several species (genomics), the development of high throughput analysis of mRNAs (transcriptomics), and the efforts to identify the protein products on a large scale (proteomics). Wide sectors of medicine are waiting for the results of these new medium and high throughput technological approaches, for example, in order to identify early markers of diseases. The object of this article is to present a biochemist's point of view on hereditary metabolic diseases (also referred to as inborn errors of metabolism), a field of medicine and research covering very diverse clinical and biochemical aspects. Significant advances will be made possible by improving the present methods of analysis of the metabolome which establishes a link between genotypes and phenotypes, an area now called metabolomics. The contribution of proteomics will be important as well but will still require some time. PMID- 15885203 TI - [Interaction of viruses with cellular response to hypoxia]. AB - Recent studies show that low oxygen tension levels in cell culture up-regulate the replication of human B19 parvovirus, Kaposi's sarcoma, and human immunodeficiency viruses as well as the expression of viral oncogenic proteins. The mechanisms of this regulation proceed with the major hypoxia-related factor, HIF-1 (hypoxia inducible factor-1). HIF-1 misregulation is implicated in the oncogenesis potential of some of these viruses. PMID- 15885204 TI - [Protein traffic in Plasmodium infected-red blood cells]. AB - To survive within erythrocytes, Plasmodium parasites have to put into place different membrane and sub-cellular compartments in order to import different nutrients and to export proteins/antigens. Infected cells pose not only a major world health risk by killing two million people per year, but also a very interesting cell biology problem, as within the erythrocyte the parasite resides inside a vacuole called the parasitophorous vacuole and as a consequence, it is separated from the blood stream by three membrane barriers, its own plasma membrane, the parasitophorous vacuole membrane and the erythrocyte plasma membrane. In spite of these three barriers the parasite is capable of secreting antigens and importing nutrients, and to do this, it has developed a complex vesicular system that extends into the red blood cell cytoplasm to the plasma membrane. Understanding how the parasite controls this extensive vesicular traffic has driven research into Plasmodium Rabs, whose potential role is discussed. PMID- 15885205 TI - [Numerical simulation of the cardiovascular system]. AB - In this article, we aim at giving a non-technical overview of some mathematical models currently used in the numerical simulation of the cardiovascular system. A hierarchy of models for blood flows in large arteries is briefly described as well as an electromechanical model for the heart. We discuss some possible applications of the numerical simulations of such models, for example the optimization of prostheses. We also address the issue of the data assimilation for the calibration of the models. PMID- 15885206 TI - [Cell microarrays and functional genomics]. AB - With the complete sequencing of the human genome, research priorities have shifted from the identification of genes to the elucidation of their function. Methods currently used by scientists to characterize gene function, such as knock out mice, are based upon loss of protein function and analysis of the resulting phenotypes to infer a potential role for the protein under scrutiny. Until now, these methods have been successful but time consuming and only a few genes at a time could be analyzed. Cell microarrays allow to simultaneously transfect thousands of different nucleic acid molecules, RNA or DNA, into adherent cells. It is then possible to analyze a large pallet of resulting phenotypes in clusters of transfected cells. We are currently manufacturing cell microarrays with collections of full-length cDNA cloned in expression vectors (gain of function analyses) or siRNA (loss of function studies) to unravel function of genes involved in differentiation and proliferation of human cells. Although there are still some technological difficulties to overcome, the potential for cell microarrays to speed up functional exploration of genomes is very promising. PMID- 15885207 TI - [DNA-arrays, a breakthrough in bacterial identification?]. AB - DNA-arrays are mainly known for their application in transcriptome analysis leading for instance to the discovery of new marker genes for diagnostics and prognostics in oncology. However, DNA arrays are also used for massively parallel analysis of DNA molecules allowing their quantification, the detection of single nucleotide polymorphisms and re-sequencing. This multi detection system is now applied to the << old >> problems of detecting and identifying bacteria in a biological sample and for the fine molecular characterization of a bacterial isolate. This new tool should serve for the diagnostic of an infection and for epidemiological studies such as those performed for the control of nosocomial infections or for the surveillance of bioterrorism attacks. DNA arrays carrying probes for 16S RNA specific of hundreds of bacterial species allow the identification of bacteria within a community by a single hybridization of amplified 16S rDNAs with universal primers and re-sequencing DNA arrays are used for multi locus sequence typing in a single step. Finally, the genome of an isolate could be characterized by DNA-arrays focused on a specific question like presence of toxin or antibiotic resistance genes. Up to now, DNA arrays are used in research laboratories for the rapid characterization at the genomic level of a strain collection, for evolutionary and population genetics studies and for the characterization of bacterial communities. Industrializing the process of DNA array construction and hybridization is now needed in order to transfer this technology to hospitals and diagnostic laboratories. PMID- 15885208 TI - [Measuring noise]. AB - High levels of noise encountered both in leisure activities and at workplaces can be somewhat annoying, but they can also cause hearing damage. In order to lessen these risks, some physical characteristics of the sound phenomenon need to be understood. The level of a sound is given in dB, a logarithmic unit in which simple addition is not available : 100 dB + 100 dB = 103 dB. The highest level of noise which can be tolerated by the human ear is considered to be 120 dB. Another component of sound characteristics is the frequency, which describes the height of a sound. The frequency is given in Hz, the human hearing field is comprised in the range of 20 to 20,000 Hz. Regarding the sensitivity of the ear, depending on the frequency, acusticians use a weighed dB, called dB(A), which takes into account a lower risk to hearing below 500 Hz and above 6 kHz. They also integrate the energy measured during a period of time to take the fluctuation of usual noise levels into account. So that currently, the levels of noise are often given in LAeq (equivalent to the level of continuous noise given in dBA). For moderate levels of noise, another weighted filter is used in sound level meters : the C curve, because low frequencies, although they are less dangerous for the ear, are more disturbing. In every day life, we sometimes have noise levels reaching 100 dB, and even 120 dB (fire alarms). Amplified music can reach 110 dBA, but a French regulation limits the output of PCPs (Walkmans) to 100 dB and the levels in concerts and discotheques to 105 dBA. At the workplace, the maximum level of noise allowed by French Law is 90 dBA for an 8 hour exposure, and 140 dB for peaks. In order to improve the protection of all workers in the EC, a recent European Directive will decrease the maximum level to 87 dBA before March 2006. PMID- 15885209 TI - [Claude Fortier: the great history of neuroendocrinology]. AB - The work of Claude Fortier is linked to the history of neuroendocrinology. Through him and his pioneer work in Montreal with Hans Selye, the << Man of stress >>, and at Laval University in Quebec City in his own laboratory, where all researchers involved in the study of the hypothalamo-hypophysial adrenal axis have been through, it is the whole saga of the search for the neuropeptide CRH (corticotropin releasing factor), and the harsh fight for the Nobel distinction that can be related. Among Claude Fortier's scientific discoveries, the feedback mechanisms of glucocorticoid hormones on brain and pituitary function, the presence of both mineralo and glucocorticoid receptors in some brain structures, and the introduction of computer science in biomedical research, can be cited. The consequences of these discoveries are illustrated in the pathologies linked to stress (anxiety, depression, addiction). Claude Fortier was not only a great figure in biomedical science, honored by several distinctions, but also an important personality in the policy of research in which he played a prominent role in Quebec medical research and allowed it to rank among the best in the world. PMID- 15885210 TI - [Resveratrol rescues mutant polyglutamine cytotoxicity in nematode and mammalian neurons]. PMID- 15885211 TI - [A matter of (fish) poison]. PMID- 15885212 TI - Airway-related vagal preganglionic neurons express brain-derived neurotrophic factor and TrkB receptors: implications for neuronal plasticity. AB - Recent evidence indicates that brain-derived neurotrophic factor (BDNF) is present in neurons and may affect neurotransmitter release, cell excitability, and synaptic plasticity via activation of tyrosine kinase B (TrkB) receptors. However, whether airway-related vagal preganglionic neurons (AVPNs) produce BDNF and contain TrkB receptors is not known. Hence, in ferrets, we examined BDNF and TrkB receptor expression in identified AVPNs using in situ hybridization and immunohistochemistry. BDNF protein levels were measured within the rostral nucleus ambiguus (rNA) region by ELISA. We observed that the subpopulation of AVPNs, identified by neuroanatomical tract tracing, within the rNA region express BDNF mRNA, BDNF protein, as well as TrkB receptor. In addition, brain tissue from the rNA region contained measurable amounts of BDNF that were comparable to the hippocampal region of the brain. These data indicate, for the first time, that the BDNF-TrkB system is expressed by AVPNs and may play a significant role in regulating cholinergic outflow to the airways. PMID- 15885213 TI - Dose-dependent neuronal injury after traumatic brain injury. AB - The Fluoro-Jade (FJ) stain reliably identifies degenerating neurons after multiple mechanisms of brain injury. We modified the FJ staining protocol to quickly stain frozen hippocampal rat brain sections and to permit systematic counts of stained, injured neurons at 4 and 24 h after mild, moderate or severe fluid percussion traumatic brain injury (TBI). In adjacent sections, laser capture microdissection was used to collect uninjured (FJ negative) CA3 hippocampal neurons to assess the effect of injury severity on mRNA levels of selected genes. Rats were anesthetized, intubated, mechanically ventilated and randomized to sham, mild (1.2 atm), moderate (2.0 atm) or severe (2.3 atm) TBI. Four or 24 h post-TBI, ten frozen sections (10 microm thick, every 15th section) were collected from the hippocampus of each rat, stained with FJ and counterstained with cresyl violet. Fluoro-Jade-positive neurons were counted in hippocampal subfields CA1, CA3 and the dentate gyrus/dentate hilus. At both 4 and 24 h post-TBI, numbers of FJ-positive neurons in all hippocampal regions increased dose-dependently in mildly and moderately injured rats but were not significantly more numerous after severe injury. Although analysis of variance demonstrated no overall difference in expression of mRNA levels for heat shock protein 70, bcl-2, caspase 3, caspase 9 and interleukin-1beta in uninjured CA3 neurons at all injury levels, post hoc analysis suggested that TBI induces increases in neuroprotective gene expression that offset concomitant increases in deleterious gene expression. PMID- 15885214 TI - Electroacupuncture attenuates morphine withdrawal signs and c-Fos expression in the central nucleus of the amygdala in freely moving rats. AB - Experimental efforts for understanding the mechanisms of electroacupuncture (EA) for opiate addiction are partially hampered by restraint stress. In unrestrained animals, it is difficult to perform EA stimulation at acupuncture points frequently selected on the four limbs. The present study was performed to evaluate the effect of EA at the acupuncture point Shen-Shu (BL.23) on morphine withdrawal signs and c-Fos expression of the amygdala in freely moving rats or restrained rats. We applied immunohistochemistry to detect c-Fos-positive nuclei. Corticosterone levels and behavioral responses were measured during EA stimulation. The needles were bilaterally inserted and fixed at BL.23, and 100-Hz electric stimulation was conducted 30 min before naloxone-precipitated withdrawal. In both freely moving rats and restrained rats, EA significantly reduced the signs of morphine withdrawal. Notably, EA stimulation in freely moving rats attenuated c-Fos expression in the central nucleus of the amygdala while EA in restrained animals increased this response. In addition, the restrained rats emitted greater levels of vocalization and facial expression than freely moving rats during EA stimulation. Corticosterone levels were also significantly higher in restrained animals after EA stimulation. The new EA paradigm demonstrated in the present study might help the analysis of certain physiological responses induced by EA that would otherwise have been hindered by restraint stress. PMID- 15885215 TI - Axonal and extracellular matrix responses to experimental chronic nerve entrapment. AB - We have analyzed the ultrastructural and histopathological changes that occur during experimental chronic nerve entrapment, as well as the immunohistochemical expression of chondroitin sulfate proteoglycan (CSPG). Adult hamsters (n = 30) were anesthetized and received a cuff around the right sciatic nerve. Animals survived for varying times (5 to 15 weeks) being thereafter perfused transcardially with fixative solutions either for immunohistochemical or electron microscopic procedures. Experimental nerves were dissected based upon the site of compression (proximal, entrapment and distal). CSPG overexpression was detected in the compressed nerve segment and associated with an increase in perineurial and endoneurial cells. Ultrastructural changes and data from semithin sections were analyzed both in control and compressed nerves. We have observed endoneurial edema, perineurial and endoneurial thickening, and whorled cell-sparse pathological structures (Renaut bodies) in the compressed nerves. Morphometrical analyses of myelinated axons at the compression sites revealed: (a) a reduction both in axon sectional area (up to 30%) and in myelin sectional area (up to 80%); (b) an increase in number of small axons (up to 60%) comparatively to the control group. Distal segment of compressed nerves presented: (a) a reduction in axon sectional area (up to 60%) and in myelin sectional area (up to 90%); (b) a decrease in axon number (up to 40%) comparatively to the control data. In conclusion, we have shown that nerve entrapment is associated with a local intraneural increase in CSPG expression, segmental demyelination, perineurial and endoneurial fibrosis, and other histopathological findings. PMID- 15885216 TI - Urocortin 2 increases c-Fos expression in topographically organized subpopulations of serotonergic neurons in the rat dorsal raphe nucleus. AB - Corticotropin-releasing factor (CRF)-related peptides modulate stress-related physiology and behavior. Some of the physiological and behavioral effects of CRF related peptides may be due to actions on CRF type 2 (CRF2) receptors modulating serotonergic systems in the dorsal raphe nucleus (DR). To determine if CRF2 receptor activation has effects on serotonergic neurons in the DR in conscious behaving rats, we gave intracerebroventricular (icv) injections of the selective CRF2 receptor agonist urocortin 2 (0, 0.01, 0.1, or 1.0 mug in 2 microl saline) to adult male Wistar rats and quantified c-Fos expression in topographically organized subpopulations of serotonergic neurons within the DR. In addition, home cage behaviors were recorded for 30 min prior to drug treatment and for 2 h following drug treatment. Two hours following drug treatment, rats were anesthetized, transcardially perfused with fixative, and brain tissues were processed for immunohistochemistry. Urocortin 2, in the absence of any effects on most behavioral endpoints studied, consistently increased c-Fos expression in subpopulations of serotonergic neurons identified by either tryptophan hydroxylase or serotonin immunostaining within specific subdivisions of the DR, particularly the dorsal region of the mid-rostrocaudal and caudal DR (-7.64, 8.18, -8.54, and -9.16 mm bregma). These studies demonstrate that urocortin 2 has selective actions on a subset of DR serotonergic neurons. Urocortin 2 actions on serotonergic systems described here may contribute to delayed behavioral effects of urocortin 2 described previously, including orexigenic, locomotor, and anxiety related effects in a variety of behavioral tests as well as potentiation of conditioned fear and induction of escape deficits in a model of learned helplessness. PMID- 15885217 TI - Might astrocytes play a role in maintaining the seizure-prone state? AB - The amygdala-kindling model is used to study complex partial epilepsy with secondary generalization. The present study was designed to (A) quantify astrocytic changes in the piriform cortex of amygdala-kindled subjects over time and (B) investigate the role that astrocytes might play in maintaining the seizure-prone state. In Study A, once the experimental subjects reached five stage 5 seizures, stimulation was stopped, and both kindled and control rats were allowed to survive for the interval appropriate to their group (7, 18, 30, or 90 days). Following each interval, the kindled and control animals were given 10 intraperitoneal injections of bromodeoxyuridine (BrdU) and sacrificed 24 h following the last injection. Significantly higher numbers of dividing astrocytes (identified by co-labeling for BrdU and to one of the astrocytic intermediate filament proteins glial fibrillary acidic protein or vimentin) were found in the kindled brains. All kindled groups had significantly higher numbers of double labeled cells on the side contralateral to the stimulation site, except for those in the 90 day survival group. In Study B, rats were implanted with chemotrodes, were kindled as in Study A, and were subsequently infused with either saline or with L alpha-AA (to lesion astrocytes) during a further 25 stimulations (1/day). L alpha-AA infused rats had significantly diminished levels of behavioral seizures, higher after discharge thresholds, lower after discharge durations, and decreased numbers of double-labeled astrocytes in piriform cortex than did saline infused rats. Together, the data indicate that astrocytes may play a role in maintaining the seizure-prone state. PMID- 15885218 TI - Re-organization of P2X3 receptor localization on epidermal nerve fibers in a murine model of cancer pain. AB - To determine whether ATP and P2X3 receptors contribute to bone-cancer pain in a mouse model, immunohistochemical techniques were used to identify whether changes in the labeling of P2X3 receptors on epidermal nerve fibers (ENFs) occurred during tumor development. C3H mice were injected with osteolytic fibrosarcoma cells in and around the calcaneus bone. These mice exhibited mechanical hyperalgesia by day 10 post-implantation as assessed using von Frey monofilaments. Biopsies of the plantar skin overlying the tumor were obtained at days 10, 14, and 18 post-implantation. Confocal images were analyzed for the number of PGP 9.5, P2X3, and CGRP immunoreactive (ir) ENFs. The overall ENF population (PGP-ir) decreased progressively over time, whereas the subsets of P2X3-ir fibers demonstrated a modest increase and CGRP-ir nerve fibers remained fairly constant. Importantly, the proportion of CGRP-ir fibers that labeled for P2X3 increased from approximately 6% in control animals to nearly 30% at day 14 following tumor cell implantation. These studies demonstrate increased expression of P2X3 receptors on CGRP-ir ENFs during tumor growth and suggest a role for ATP in cancer-related pain. PMID- 15885219 TI - Proteomic identification of proteins oxidized by Abeta(1-42) in synaptosomes: implications for Alzheimer's disease. AB - Protein oxidation has been implicated in Alzheimer's disease (AD) and can lead to loss of protein function, abnormal protein turnover, interference with cell cycle, imbalance of cellular redox potential, and eventually cell death. Recent proteomics work in our laboratory has identified specifically oxidized proteins in AD brain such as: creatine kinase BB, glutamine synthase, ubiquitin carboxy terminal hydrolase L-1, dihydropyrimidase-related protein 2, alpha-enolase, and heat shock cognate 71, indicating that a number of cellular mechanisms are affected including energy metabolism, excitotoxicity and/or synaptic plasticity, protein turnover, and neuronal communication. Synapse loss is known to be an early pathological event in AD, and incubation of synaptosomes with amyloid beta peptide 1-42 (Abeta 1-42) leads to the formation of protein carbonyls. In order to test the involvement of Abeta(1-42) in the oxidation of proteins in AD brain, we utilized two-dimensional gel electrophoresis, immunochemical detection of protein carbonyls, and mass spectrometry to identify proteins from synaptosomes isolated from Mongolian gerbils. Abeta(1-42) treatment leads to oxidatively modified proteins, consistent with the notion that Abeta(1-42)-induced oxidative stress plays an important role in neurodegeneration in AD brain. In this study, we identified beta-actin, glial fibrillary acidic protein, and dihydropyrimidinase-related protein-2 as significantly oxidized in synaptosomes treated with Abeta(1-42). Additionally, H+-transporting two-sector ATPase, syntaxin binding protein 1, glutamate dehydrogenase, gamma-actin, and elongation factor Tu were identified as increasingly carbonylated. These results are discussed with respect to their potential involvement in the pathogenesis of AD. PMID- 15885220 TI - Normal hypothalamo-pituitary-adrenal axis function in a rat model of peripheral neuropathic pain. AB - Chronic pain conditions such as rheumatoid arthritis and fibromyalgia are associated with profound hypothalamo-pituitary-adrenal (HPA) axis dysfunction which may exacerbate symptoms of chronic pain. HPA axis dysfunction has also been well documented in animal models of chronic inflammatory pain. However, the role of the HPA axis in animal models of neuropathic pain is currently unknown. Rats with a chronic constriction injury (CCI) of the sciatic nerve that developed marked mechanical allodynia and hyperalgesia of the injured hindpaw were used to determine basal and stimulatory levels of HPA axis activity. Plasma ACTH and corticosterone levels were increased significantly (P < 0.05) in CCI rats after 20 min restraint stress compared with baseline; however, the magnitude of the increase was no different from sham rats. Furthermore, the temporal profile of ACTH release over the 60 min period after termination of restraint was similar between CCI and sham rats suggesting normal glucocorticoid-mediated feedback. Restraint stress also significantly increased (P < 0.05) expression of the immediate early genes c-Fos and FosB within the hypothalamic PVN to a similar extent in CCI and sham rats. Within the parvocellular PVN basal expression of both CRF and AVP mRNA was no different between CCI and sham rats; restraint stress induced a significant 2.5 fold increase (P < 0.05) in CRF mRNA expression in sham rats only. These results suggest that, in contrast to inflammatory immune mediated pain models where HPA axis function is profoundly altered, in the CCI model of neuropathic pain, basal HPA axis function is unchanged. Furthermore, the HPA axis responds normally to a novel stressor in the face of ongoing nociceptive input, a stimulus known to activate the HPA axis. PMID- 15885221 TI - Contribution of mitogen-activated protein kinases to NMDA-induced neurotoxicity in the rat retina. AB - We examined the contributions of the mitogen-activated protein kinases (MAPKs) family [extracellular signal-regulated kinase (ERK), p38 kinase (p38), and c-Jun N-terminal kinase (JNK)] to N-methyl-D-aspartate (NMDA)-induced neurotoxicity in the rat retina. Detection of apoptotic cell death in the retinal ganglion cell layer (RGCL) and the inner nuclear layer (INL) by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) staining began 6 h after intravitreal NMDA (100 nmol) injection and continued to increase thereafter. Western blot analysis showed that phosphorylated MAPKs (p-MAPKs) were expressed in the retina following a temporal manner: maximal expression of phosphorylated ERK (p-ERK) at 1 h, maximal expression of phosphorylated p38 (p p38) at 6 h, and beginning of phosphorylated JNK (p-JNK) significant increase at 6 h after injection. An immunohistochemical/TUNEL co-localization study showed that p-JNK- and p-p38-positive cells in the RGCL were frequently TUNEL-positive, whereas few p-ERK-positive cells were TUNEL-positive. Moreover, co-injection of inhibitors for JNK (0.2 nmol SP600125) and/or p38 (2.0 nmol SB203580) with NMDA was effective in ameliorating NMDA-induced apoptotic cell loss in the RGCL 12 h after injection, as shown by TUNEL-positive cell counts. These inhibitors also protected the inner retina as shown by morphometric studies such as cell counts in the RGCL and measurement of the IPL thickness 7 days after injection. On the other hand, an ERK inhibitor (2.0 nmol U0126) did not suppress NMDA-induced cell death in the RGCL nor thinning of the IPL. These findings suggest that JNK and p38 are proapoptotic in NMDA-induced cell death in the RGCL, but not ERK. PMID- 15885222 TI - Afferent pathways to the region of the vestibular nuclei that participates in cardiovascular and respiratory control. AB - Prior experiments have shown that a region of the medial and inferior vestibular nuclei contributes to cardiovascular and respiratory regulation. In addition to labyrinthine inputs, the majority of neurons in this region of the vestibular nuclei receive signals from the skin, muscle, and viscera, although the pathways conveying these nonlabyrinthine inputs to the vestibular nucleus neurons are unknown. To gain further insight into the afferent pathways to this functionally distinct subdivision of the vestibular complex, we combined monosynaptic mapping with viral transneuronal tracing in the ferret. First order afferent projections were defined by retrograde transport of the beta-subunit of cholera toxin (CTbeta), and the extended polysynaptic circuitry was defined in the same animals by injection of a recombinant of pseudorabies virus Bartha (PRV) into the contralateral vestibular nuclei. Neurons containing CTbeta or infected by retrograde transneuronal transport and replication of PRV were distributed throughout the spinal cord, but were 10 times more prevalent in the cervical cord than the lumbar cord. The labeled spinal neurons were most commonly observed in Rexed's laminae IV-VI and the dorsal portions of laminae VII-VIII. Both the CTbeta and PRV injections also resulted in labeling of neurons in all four vestibular nuclei, the prepositus hypoglossi, the reticular formation, the inferior olivary nucleus, the medullary raphe nuclei, the spinal and principal trigeminal nuclei, the facial nucleus, and the lateral reticular nucleus. Following survival times >/=3 days, PRV-infected neurons were additionally present in nucleus solitarius and the gracile and cuneate nuclei. These data show that an anatomical substrate is present for somatosensory and visceral inputs to influence the activity of cells in the autonomic region of the vestibular nuclei and suggest that these signals are primarily transmitted through brainstem relay neurons. PMID- 15885223 TI - Changes in glutamate receptor subunit 4 expression in the deafferented olfactory bulb of zebrafish. AB - The distribution of ionotropic glutamate receptor subunit 4 (iGluR4) was examined in both normal and deafferented olfactory bulbs of adult zebrafish, Danio rerio. With the exception of the olfactory nerve layer, there was extensive labeling with antibodies to iGluR4 in the olfactory bulbs, specifically in juxtaglomerular cell bodies and their processes. These results are consistent with previous work, which has suggested differential distribution of glutamate receptors in the vertebrate olfactory system. Analysis of bulbs following olfactory organ removal revealed a significant loss of iGluR4 immunoreactivity by 24 h post deafferentation. At 48 h after denervation, iGluR4 labeling had returned to normal levels and was retained through 3 weeks post-surgery. Thus, afferent input plays a role in reduced labeling of this protein immediately following injury, but return of immunoreactivity can occur even without sensory innervation. PMID- 15885224 TI - Immunohistochemical co-expression of carbonic anhydrase II with Kv1.4 and TRPV1 in rat small-diameter trigeminal ganglion neurons. AB - The co-expression of carbonic anhydrase II (CAII) with the voltage-gated potassium channel subtype 1.4 (Kv1.4) or the vanilloid receptor (TRPV1) was examined in adult rat trigeminal ganglion (TG) neurons by using the immunofluorescence method. The small-diameter Kv.1.4-positive TG neurons co expressed CAII immunoreactivity (47%). Most TRPV1-positive TG neurons (79%) had the CAII immunoreactivity, but showed a lack of immunoreactivity for a neurofilament protein (NF200), a maker of large TG neurons with myelinated axons. The fact that CAII-immunoreactive TG neurons revealed a common expression of both Kv1.4 and TRPV1 leads us to suggest that CAII may be one of the nociceptive neuronal markers. PMID- 15885225 TI - Learning-induced plasticity of cortical representations does not affect GAD65 mRNA expression and immunolabeling of cortical neuropil. AB - Two forms of glutamic acid decarboxylase (GAD) are present in inhibitory neurons of the mammalian brain, a 65-kDa isoform (GAD65) and a 67-kDa isoform (GAD67). We have previously found that GAD67 is upregulated during learning-dependent plasticity of cortical vibrissal representations of adult mice. After sensory conditioning involving pairing stimulation of vibrissae with a tail shock, the increase in mRNA expression and density of GAD67-immunoreactive neurons was observed in barrels representing vibrissae activated during the training. In the present study, using the same experimental model, we examined GAD65 mRNA and protein levels in the barrel cortex. For this purpose, we used in situ hybridization and immunohistochemistry. No changes in the level of GAD65 mRNA expression were detected after the training. The pattern of GAD65 mRNA expression was complementary to that observed for GAD67. Immunocytochemical analysis found no changes in immunolabeling of neuropil of the barrels representing the vibrissae activated during the training. The results show that, in contrast to GAD67, cortical plasticity induced by sensory learning does not affect the expression of GAD65. PMID- 15885226 TI - RNA helicases: regulators of differentiation. AB - RNA helicases are highly conserved enzymes that utilize the energy derived from NTP hydrolysis to modulate the structure of RNA. RNA helicases participate in all biological processes that involve RNA, including transcription, splicing and translation. Based on the sequence of the helicase domain, they are classified into families, such as DDX and DHX families of human RNA helicases. The specificity of RNA helicases to their targets is likely due to several factors, such as the sequence, interacting molecules, subcellular localization and the expression pattern of the helicases. There are several examples of the involvement of RNA helicases in differentiation. Human DDX3 has two closely related genes designated DDX3Y and DDX3X, which are localized to the Y and X chromosomes, respectively. DDX3Y protein is specifically expressed in germ cells and is essential for spermatogenesis. DDX25 is another RNA helicase which has been shown to be required for spermatogenesis. DDX4 shows specific expression in germ cells. The Drosophila ortholog of DDX4, known as vasa, is required for the formation of germ cells and oogenesis by a mechanism that involves regulating the translation of mRNAs essential for differentiation. Abstrakt is the Drosphila ortholog of DDX41, which has been shown to be involved in visual and CNS system development. DDX5 (p68) and its related DDX17 (p72) have also been implicated in organ/tissue differentiation. The ability of RNA helicases to modulate the structure and thus availability of critical RNA molecules for processing leading to protein expression is the likely mechanism by which RNA helicases contribute to differentiation. PMID- 15885227 TI - Comparison of three strategies for myocardial protection during coronary artery bypass graft surgery based on markers of cardiac damage. AB - OBJECTIVES: To evaluate myocardial damage during coronary artery bypass grafting using three different intermittent cardioplegia and then measuring cTnI and CKMBm release. DESIGN AND METHODS: Forty-two patients belonging to the hypothermic crystalloid (n = 16), hypothermic (n = 13), and normothermic blood (n = 13) groups were collected when removing the aortic cross-clamp (t = 0) and after 4, 12, 24 and 48 h. For each patient, cumulative cTnI and CKMBm release was calculated as the five measurement mean. There were no significant preoperative and operative differences in the three groups. RESULTS: In the normothermic group, cTnI mean values at 4, 12, and 24 h were significantly lower than those in both hypothermic groups; moreover, CKMBm mean values were higher at 4, 12, and 24 h in the hypothermic crystalloid group and at 4 and 12 h in the hypothermic blood group than in the normothermic group. In the normothermic group, the area under the curve of the release of both markers was significantly lower than in the hypothermic groups. No significant difference was reported in the release of both markers in hypothermic groups. CONCLUSIONS: A strategy of normothermic cardioplegia seems to preserve myocardium better than hypothermic cardioplegia. PMID- 15885228 TI - Comparison of apolipoprotein B and plasma lipids as targets for lipid lowering treatment. AB - OBJECTIVE: To assess the discrepancies between LDL-cholesterol and apo B targets in patients at risk for vascular disease; namely, those with diabetes or hypertriglyceridemia and those with triglycerides <1.7 mmol/L and normal glucose homeostasis. METHODS: Lipid clinic patients were divided into two groups: Group 1 consisted of 182 patients whose triglyceride levels were >1.7 mmol/L or who had impaired fasting glucose or type 2 diabetes. In Group 2, there were 42 patients with triglycerides <1.7 mmol/L and normal glucose homeostasis. LDL-cholesterol and apo B were estimated during lipid clinic visits with patients on appropriate lipid lowering therapy. RESULTS: 46% of the patients in Group 1 who reached the high risk LDL-cholesterol target of <2.5 mmol/L did not reach apo B target of <0.9 g/L, while in Group 2, only 19% of those who reached the LDL-cholesterol target had apo B >0.9 g/L. CONCLUSION: Our findings demonstrate that a large percentage of patients with hypertriglyceridemia or impaired glucose tolerance, treated with lipid lowering agents, reach the LDL-cholesterol but not the apo B treatment targets. PMID- 15885229 TI - Osmolality revisited--deriving and validating the best formula for calculated osmolality. AB - OBJECTIVE: To derive a formula that can be used (i) to calculate osmolality in normal patients as well as those that are hyperglycemic and intoxicated, and (ii) to predict the presence of unexplained compounds with the osmol gap calculation in the presence and absence of ethanol. DESIGN AND EXPERIMENTS: We performed in vitro experiments to determine the relationship of serum osmolality with sodium, potassium, urea, glucose, ethanol, methanol, and ethylene glycol. Several formulas were then tested for their validity in predicting osmolality in normal individuals. Finally, we assessed whether these formulas would allow us to calculate the osmolality gap (OG) that may be indicative of the presence of other osmotically active compounds. The OG calculation was done both in the presence and absence of ethanol. In this way, the OG should be able to detect compounds like methanol and ethylene glycol even in the presence of ethanol which is easily measured and is very often present in the above-named poisonings. RESULTS: Experimental results show that glucose, ethanol, methanol, and ethylene glycol need factors of 1.15, 1.20, 1.07, and 1.00, respectively, to accurately predict osmolality. The factors for glucose and ethanol were then validated in normal subjects as well as in a large patient database. The formulas below predicted osmolality very well in patients whether ethanol was present or not. All concentrations are expressed in mmol/L. The mean osmol gap for healthy subjects without ethanol present was 0.77 +/- 3.80 mosM/kg with the reference interval being -6.68 to 8.23 mosM/kg for formula 1 and -8.04 to 6.50 mosM/kg for formula 2. The mean osmol gap (OG) in patients who had ethanol present was 1.22 +/- 5.32 for formula 1 and -0.2 +/- 5.0 for formula 2. CONCLUSIONS: This study shows that factors of 1.20 and 1.15 have to be applied to ethanol and glucose to allow for accurate calculation of osmolality and osmolality gap. There were insufficient patient data to verify the factors for methanol and ethylene glycol. PMID- 15885230 TI - The value of biliary amylase and Hepatocarcinoma-Intestine-Pancreas/Pancreatitis associated Protein I (HIP/PAP-I) in diagnosing biliary malignancies. AB - BACKGROUND/OBJECTIVES: Elevated concentrations of Hepatocarcinoma-Intestine Pancreas/Pancreatitis-associated Protein I (HIP/PAP-I) in pancreatic juice have been reported in patients with pancreatic adenocarcinoma and have been considered as a promising tumor marker. This study was conducted to investigate whether biliary HIP/PAP-I can be used in the differential diagnosis of the cause of biliary obstruction. METHODS: Bile was obtained from patients with bile duct obstruction on the day of biliary drainage. The etiology of biliary obstruction included gallstones (n = 131), pancreatic cancer (n = 32), cholangiocarcinoma (n = 47), papilla Vater cancer (n = 13), hepatocellular carcinoma (n = 4) and metastatic cancer (n = 16). In addition to HIP/PAP-I, the samples were analyzed for amylase to check for the presence of pancreaticobiliary reflux. RESULTS: The biliary concentration of HIP/PAP-I was not statistically different between patients with gallstones (median, 9.70 ng/mL; interquartile range [IQR] 1.80 45.75) and cancers (median, 12.70 ng/mL; IQR, 3.85-36.75), P > 0.05. However, the amylase activity in the bile was markedly elevated in patients with gallstones (median, 228 U/L; IQR, 40-1965), compared to those with cancer (median, 32 U/L; IQR; 30-176), P < 0.001. The area under the ROC curve of amylase was 0.751 (95% CI: 0.69 to 0.81). At a cut-off value of 46 U/L, the biliary amylase distinguished patients with malignant obstruction from those with benign obstruction with a sensitivity of 66% and a specificity of 74%. CONCLUSIONS: Our data suggest that the biliary HIP/PAP-I measurement is not useful for differentiating causes of biliary obstruction. The divergent extent and duration of biliary obstruction caused by neoplasm and gallstones may contribute to the significant difference in the amylase activity in bile. Thus, amylase in bile represents a candidate marker in the differential diagnosis of the cause of biliary obstruction. PMID- 15885231 TI - Adrenomedullin and total nitrite levels in children with acute rheumatic fever. AB - OBJECTIVE: To investigate the levels of adrenomedullin (AM) and total nitrite, a stable product of nitric oxide (NO), in children with acute rheumatic fever (ARF). DESIGN AND METHODS: Eleven children with ARF were investigated in comparison with 14 healthy controls. Adrenomedullin was detected by HPLC, while total nitrite was quantitated by the Griess reaction. RESULTS: Plasma urinary AM and total nitrite levels were significantly higher in children with ARF, irrespective of whether they were in the acute or convalescent phase of disease. Plasma AM (pmol/mL) levels were 49.19 +/- 3.23 in the acute phase, 44.52 +/- 4.26 in the convalescent phase, 35.49 +/- 3.43 in controls, and urinary AM excretion (pmol/mg creatinine) was 43.45 +/- 18.40 in the acute phase, 32.38 +/- 15.37 in the convalescent phase, and 24.84 +/- 11.38 in controls. Plasma total nitrite levels (mumol/L) were 75.37 +/- 13.13 in the acute phase, 59.81 +/- 12.78 in the convalescent phase, and 41.09 +/- 10.27 in controls. Urinary total nitrite excretion (mumol/mg creatinine) was 3.82 +/- 1.56 in the acute phase, 2.15 +/- 0.58 in the convalescent phase, and 1.33 +/- 0.61 in controls. The differences were statistically significant for all (P < 0.05). CONCLUSION: This study considered that AM and NO may have a role in the immunoinflammatory process of ARF. PMID- 15885232 TI - The quantification of cytochrome P-450 (CYP 3A4) mRNA in the blood of patients with viral liver diseases. AB - OBJECTIVE: We quantified cytochrome P-450 (CYP) 3A4 mRNA in the blood and liver of patients with viral liver diseases to determine whether CYP 3A4 expression is related to disease progression. DESIGN AND METHODS: Total RNA was extracted from 10 mL of blood from 12 normal volunteers, from 6 patients with acute hepatitis, 17 with chronic hepatitis, 12 with liver cirrhosis, and 16 with hepatocellular carcinoma. Total RNA from 1 mg of liver tissue was extracted simultaneously in 10 patients. CYP 3A4 mRNA was quantified by competitive reverse-transcription polymerase chain reaction and expressed as log copies/microliter. RESULTS: The CYP 3A4 mRNA titer in blood correlated with that of the liver (r = 0.65, P < 0.05). The CYP 3A4 mRNA titer was 1.6 +/- 0.4 in normal controls, 1.0 +/- 0.5 in acute hepatitis, 0.7 +/- 0.2 in chronic hepatitis, 0.5 +/- 0.2 in liver cirrhosis, 0.5 +/- 0.2 in hepatocellular carcinoma, and decreased with progression of liver disease (P < 0.05). CONCLUSION: These data suggest that the CYP 3A4 mRNA level in blood relates to progression of liver disease. PMID- 15885233 TI - A case control study of glycoprotein status in ovarian carcinoma. AB - OBJECTIVES: Ovarian cancer is the leading cause of death due to gynecological malignancies. The aim of our study was to investigate the status of circulating glycoprotein levels in ovarian cancer patients. DESIGN AND METHODS: Thirty ovarian cancer patients and an equal number of age-matched, apparently healthy subjects as controls were involved in the study. Glycoprotein levels, as indicated by the concentration of plasma total sialic acid, protein-bound hexoses, hexosamine and fucose were estimated in circulation of both the ovarian cancer patients and controls. RESULTS: Significantly elevated levels of plasma total sialic acid, protein-bound hexoses, hexosamine and fucose were observed in ovarian cancer patients as compared to the apparently healthy controls. CONCLUSION: Plasma total sialic acid, protein-bound hexoses, hexosamine and fucose in the circulation of ovarian cancer patients are markedly elevated and the increase in these carbohydrate moieties of glycoproteins reflect the stage of cancer and may be an additional tool in the diagnosis and prognosis of ovarian carcinoma. PMID- 15885234 TI - Thymosin alpha 1 attenuates lipid peroxidation and improves fructose-induced steatohepatitis in rats. AB - OBJECTIVES: The aim of this study was to investigate the effects of thymosin alpha(1) (Talpha(1)) in rats having fructose-induced steatosis. Fructose leads to experimental steatosis in the liver by exerting its effect on some components of the oxidant/antioxidant system, and on several cytokines (interleukin-1beta, -2, and -6) in blood. METHODS: Twenty-four rats at random were divided into three groups (each group containing eight animals); the control group (C), which received a purified diet; the high-fructose-fed group (F); and the high-fructose fed and Talpha(1) injected group (F + T). After the experimental period of 10 days, liver lipid peroxidation and antioxidant status, and blood IL-1beta, IL-2, and IL-6 levels were quantified. RESULTS: In comparison with the C group, the F group had a higher nitric oxide (NO) level, xanthine oxidase (XO) activity, and lipid peroxidation, as indicated by concentrations of thiobarbituric acid reactive substances (TBARS), and lower superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the liver. In the F + T group, these markers were near the values of the control group. In addition, increased IL-1beta and IL 6 levels were kept at near to normal levels with treatment of Talpha(1), but not IL-2 levels. In the F group, the most consistent findings in the histologic sections of liver tissues were the macrovesicular and microvesicular steatosis. Talpha(1) treatment protected the majority of the liver cells, while minimal macrovesicular and microvesicular steatosis was observed in the remaining cells. CONCLUSIONS: These results show that a high-fructose diet in rats leads to hepatic steatosis and a defect in the free radical defense system, and that treatment of Talpha(1) may improve these biochemical and morphologic changes in the fructose-fed rat livers. PMID- 15885235 TI - Evaluation of the turnaround time of an integrated preanalytical and analytical automated modular system in a medium-sized laboratory. AB - OBJECTIVES: Evaluation of an integrated Modular Preanalytics (MPA) and Modular Analytics SWA (MA) system (Roche Diagnostics) during continuous batch processing. DESIGN AND METHODS: A total of 1000 blood specimen tubes was processed and tested in a batch-wise fashion, according to two different specimen input conditions (Study 1 and Study 2). The resulting turnaround time of the system was assessed. RESULTS: Study 1 tubes were centrifuged in the MPA. The preanalytical time rose steadily from 14 to 28 min, and after sample 315, it showed minimal variation. The analytical time remained almost constant. In Study 2, tubes were centrifuged before being processed in the MPA. The preanalytical time increased from 4 to 19 min, and the analytical time increased similarly. The turnaround time in Study 1 was 132 min and in Study 2 was 108 min. CONCLUSION: Centrifugation in the MPA slightly increased the turnaround time. Nevertheless, the labor associated with specimen processing was reduced and the efficiency of the laboratory was improved. PMID- 15885236 TI - Effects of some hematological parameters on whole blood tacrolimus concentration measured by two immunoassay-based analytical methods. AB - OBJECTIVES: Tacrolimus (FK506) is a potent immunosuppressive drug used for prevention of rejection following transplantation. Several methods including immunoassays have been used for monitoring tacrolimus levels. The purpose of the present study was to compare the effects of various hematological parameters on whole blood tacrolimus concentrations which were measured with two different analytical methods, namely the microparticle enzyme immunoassay (MEIA II) and enzyme multiplied immunoassay technique (EMIT). DESIGN AND METHODS: The effects of hematological variables, namely hematocrit (Htc), hemoglobin (Hb), red blood cell (RBC), mean cell volume (MCV), mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), red cell distribution width (RDW) and platelet (PLT) counts on tacrolimus concentrations (n = 2430 measurements) measured with EMIT (n = 1171) and MEIA II (n = 1259) methods in whole blood samples from kidney or liver or combined kidney-pancreas transplant patients (n = 162) during a 2 year post-transplantation period were compared. RESULTS: The whole blood tacrolimus concentrations measured with MEIA II method were affected much more significantly by hematological parameters than those measured with EMIT method. In MEIA II method, RDW (r = 0.479, P < 0.01) showed a stronger correlation with tacrolimus concentration than Htc (r = -0.239, P < 0.01) in all patients. A negative significant correlation (r = -0.468, P < 0.01) was also observed between the Htc and tacrolimus concentration in patients with Htc values < or =25% in MEIA II method. CONCLUSIONS: The results of the present study suggest that EMIT method might be preferred to MEIA II in determination of whole blood tacrolimus concentrations in anemic transplant patients. For better therapeutic drug monitoring, physicians should be aware of these assay differences. Evaluation of hematologic factors that affect the whole blood concentrations of tacrolimus may be helpful in deciding the dosage of this drug. PMID- 15885237 TI - Development of a sensitive and specific enzyme-linked immunosorbent assay for thymosin beta15, a urinary biomarker of human prostate cancer. AB - OBJECTIVES: In tissue-based assays, thymosin beta15 (Tbeta15) has been shown to correlate with prostate cancer (CaP) malignancy and with future recurrence. To be clinically effective, it must be shown that Tbeta15 is released by the tumor into body fluids in detectable concentrations. Toward this end, we have worked to develop a quantitative high-throughput assay that can accurately measure clinically relevant concentrations of Tbeta15 in human urine. DESIGN AND METHODS: Sixteen antibodies were raised against recombinant Tbeta15 and/or peptide conjugates. One antibody, having stable characteristics over the wide range of pH and salt concentrations found in urine and minimal cross-reactivity with other beta thymosins, was used to develop a competitive enzyme-linked immunosorbent assay (ELISA). Urinary Tbeta15 concentration was determined for control groups; normal (N = 52), prostate intraepithelial neoplasia (PIN, N = 36), and CaP patients; untreated (N = 7) with subsequent biochemical failure, radiation therapy (N = 17) at risk of biochemical recurrence. RESULTS: The operating range of the competition ELISA fell between 2.5 and 625 ng/mL. Recoveries exceeded 75%, and the intra- and inter-assay coefficients of variability were 3.3% and 12.9%, respectively. No cross-reactivity with other urine proteins was observed. A stable Tbeta15 signal was recovered from urine specimens stored at -20 degrees C for up to 1 year. At a threshold of 40 (ng/dL)/mug protein/mg creatinine), the assay had a sensitivity of 58% and a specificity of 94%. Relative to the control groups, Tbeta15 levels were greater than this threshold in a significant fraction of the CaP patients (P < 0.001), including 5 of the 7 patients who later experienced PSA recurrence. CONCLUSIONS: We have established an ELISA that is able to detect Tbeta15 at clinically relevant concentrations in urine from patients with CaP. The assay will provide a tool for future clinical trials to validate urinary Tbeta15 as a predictive marker for recurrent CaP. PMID- 15885238 TI - Limited preanalytical requirements for insulin measurement. AB - OBJECTIVES: To evaluate the stability of insulin in serum and plasma under conditions relevant for the transport of samples from outpatient clinics to a centralized laboratory. DESIGN AND METHODS: Venous blood samples were taken from 15 volunteers and processed either immediately or after storage at room temperature for 24 h. The effect of EDTA or sodium fluoride on insulin results was investigated. RESULTS: Insulin was found to be stable in EDTA-containing tubes at room temperature for at least 24 h, whereas significantly lower concentrations were found if samples without additives or potassium fluoride as an additive were used. CONCLUSIONS: If shipment of patients' samples for insulin measurement from an outpatient clinic to a centralized laboratory can be performed within 24 h, centrifugation and freezing of plasma is not required. These findings facilitate a widespread application of insulin measurements to characterize and follow-up individual insulin sensitivity. PMID- 15885239 TI - Spectrum of beta-thalassemia mutations in North Indian states: a beta-thalassemia trait with two mutations in cis. AB - OBJECTIVES: To study the prevalence of beta-thalassemia mutations in two North Indian states. DESIGN AND METHODS: Mutation(s) were analyzed in 62 patients using 10 sets of allele-specific primers. RESULTS: Four mutations [IVS1 nt5 (G --> C), F.S 8/9 (+G), F.S 41/42 (-TCTT) and del 619 bp] were found to constitute 94.1% of the total alleles studied. Prevalence of IVS1 nt5 (G --> C) was the highest, closely followed by F.S 8/9 (+G). The latter was thus unusually high and not consistent with earlier reports. Two mutations in cis were detected in one carrier of the thalassemia trait and in his father, thus signifying the possibility of vertical transmission of mutations in a single beta-globin allele. CONCLUSIONS: F.S 8/9 (+G), as well as IVS1 nt5 (G --> C), are the major mutations in Indian states; however, the possibility of multiple mutations in cis in a fetus with the thalassemia trait has to be considered in a prenatal screening program. PMID- 15885240 TI - A modified conformation sensitive gel electrophoresis (CSGE) method for rapid and accurate detection of low density lipoprotein (LDL) receptor gene mutations in Familial Hypercholesterolemia. AB - OBJECTIVES: A conformation sensitive gel electrophoresis (CSGE) method was modified for detection of LDL receptor gene mutations. DESIGN AND METHODS: Usage of a temperature gradient against running time was tested with 33 identified mutations in the LDL receptor gene. Most of the mutations were missense. RESULTS: 32 mutations were detected using this method (sensitivity: 97%). The duration required for running the test for each of the exons was reduced to 3-4 h. CONCLUSIONS: This modified CSGE method may be used as a screening procedure in genetic diseases where a variety of mutations may cause illness. PMID- 15885241 TI - Heterophile antibodies produce spuriously elevated concentrations of cardiac Troponin I in patients with Legionella pneumophila. AB - We found an unusually high positive rate for cTnI in patients recently infected with Legionella pneumophila. The aim of this study was to examine the possible origin of increased cTnI levels and to test if it could be associated with the immune response to legionellosis. The cTnI was above the cut point in 46.7% of patients infected with legionellosis when measured with reagent lot number RF421A. A strong correlation between high cTnI measurements and positive serologic values for legionellosis was found. With a revised formulation of cTnI reagent, lot number RF421C, the positive rate decreased by over 10-fold to 3.3%. We conclude that the revised lot of cTnI reagent minimized interference by heterophilic antibodies produced in response to legionellosis. PMID- 15885242 TI - Imprecise control of energy intake: absence of a reduction in food intake following overfeeding in young adults. AB - The objective was to examine the extent to which overfeeding reduces spontaneous food intake in humans. Twelve normal-weight adults participated in the three stage study. During the 14 day baseline period and 21 day recovery period, food intake was consumed ad libitum, beyond a minimum 5 MJ (1200 kcal) basal diet. During the 13 day period of overfeeding, each subject consumed 35% more energy than they consumed at baseline. Overfeeding resulted in a weight gain of 2.3+/ 0.37 kg, (p<0.0001), approximately half the weight gain was determined to be fat (1.2+/-0.19 kg, p<0.0001) by underwater densitometry. Following overfeeding, mean daily caloric intake was not significantly suppressed returning immediately to baseline values. Despite normal energy intake, participants lost 1.3+/-0.24 kg of body weight (p<0.0001), of which 0.75+/-0.15 kg (p<0.0001) was fat. These results indicated that (1) the physiological control of eating behavior in humans is not the major mechanism responsible for the recovery of body weight following a period of overfeeding and (2) an increase in energy expenditure of 1.28 MJ (307 kcal)/day or about 14% was required to account for the weight loss following overfeeding. PMID- 15885243 TI - Influence of personality traits on plasma levels of cortisol and cholesterol. AB - The literature reports many organic malfunctions that are associated with elevated plasma cortisol and cholesterol levels. The present investigation was concerned with the influence of personality on plasma levels of cortisol and cholesterol. To that effect these variables were determined in a group of 20 subjects who answered the Big-Five Inventory for measurements of personality traits. It was found that: among the 5 personality traits, extraversion was positively correlated to plasma levels of cortisol and cholesterol while the correlation was negative for neuroticism. The positive correlation between extraversion and plasma cortisol and cholesterol, as well as with the responses to stress as shown in a previous study, are similar to findings previously reported on type A individuals. Further studies are needed with a larger group of subjects to conclude to a direct causal relationship between extraversion and the high levels of plasma cortisol and cholesterol, or a predisposition to some organic malfunctions as is the case for type A. PMID- 15885244 TI - Differential fos activation in virgin and lactating mice in response to an intruder. AB - Lactating (L) mice display fierce aggression towards novel, male mice, while virgin (V) mice do not. This study compares patterns of brain activation in V and L mice in response to a novel intruder using immunohistochemical detection of Fos (Fos-IR). Animals were sampled 120 min after either a sham or real 10 min test with a male intruder. L mice were aggressive towards intruders, but V mice were not. In general, Fos-IR for both groups increased with exposure to an intruder, with L mice showing higher increases in Fos-IR than V mice. In only medial preoptic nucleus and ventral portion of bed nucleus of stria terminalis (BNST) was Fos-IR significantly increased in both groups with testing. In V mice, testing resulted in Fos-IR increases in an additional 10 regions examined that did not reach significance in L mice, including lateral septum, lateral and medial preoptic areas, and anterior hypothalamus. Fos-IR also increased with testing in nine regions unique to L mice, including the mitral and granular layers of accessory olfactory bulb, regions of the amygdala, dorsal BNST, and caudal portions of the hypothalamic attack area. These increases in Fos-IR with testing suggest alterations in the circuitry governing response to pheromonal cues and imply some commonalities between the circuitries governing maternal aggression and intermale aggression. These results support the hypothesis that pregnancy and lactation induce substantial changes in brain circuitry and function; changes that enable maternal defense of offspring by altering the neural response to an intruder male. PMID- 15885245 TI - Neuroendocrine and cytokine profile of chronic mild stress-induced anhedonia. AB - A bidirectional relationship exists between depression and cardiovascular disease. Patients with major depression are more likely to develop cardiac events, and patients with myocardial infarction and heart failure are more likely to develop depression. A feature common to both clinical syndromes is activation of proinflammatory cytokines and stress hormones, including the hypothalamic pituitary-adrenal axis and the renin-angiotensin-aldosterone system. In the present study we examined the hypothesis that exposure to chronic mild stress (CMS), an experimental model of depression that induces anhedonia in rats, is sufficient to activate the production of proinflammatory cytokines and stress hormones that are detrimental to the heart and vascular system. Four weeks of exposure of male, Sprague-Dawley rats to mild unpredictable environmental stressors resulted in anhedonia which was operationally defined as a reduction in sucrose intake without a concomitant effect on water intake. Humoral assays indicated increased plasma levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), plasma renin activity, aldosterone, and corticosterone in the CMS exposed rats. Tissue TNF-alpha and IL-1beta were increased in the hypothalamus, and TNF-alpha was increased in the pituitary gland. These humoral responses to CMS, associated with anhedonia as an index of depression in the rat, are likely to be associated with neurohumoral mechanisms that may contribute to adverse cardiac events. The findings provide a basis for examining more directly the interactions among the central, endocrine, and immune systems in depression associated with heart disease. PMID- 15885246 TI - The effects of SB 224289 on anxiety and cocaine-related behaviors in a novel object task. AB - Cocaine facilitates dopamine transmission from ventral tegmental area (VTA) neurons that project to nucleus accumbens (NAcc), and previous experiments suggest that serotonin-1B (5-HT1B) receptors are involved in this effect. Specifically, activation of 5-HT1B receptors in VTA during cocaine exposure increases dopamine release in NAcc and enhances cocaine-induced locomotor activity, reward, and reinforcement. Thus, it is reasonable to hypothesize that blocking 5-HT1B activity may have the opposite effect. To investigate this hypothesis, SB 224289, a highly selective 5-HT1B antagonist, was used to block this receptor. In an open field/novel object exploration test, SB 224289 reduced cocaine-induced locomotion. However, SB 224289 also increased anxiety-like behavior, both alone and in combination with cocaine. This experiment gives evidence that 5-HT1B antagonists may reduce some of the behavioral effects of cocaine, but may have negative effects on anxiety as well. PMID- 15885247 TI - Effect of driving duration and partial sleep deprivation on subsequent alertness and performance of car drivers. AB - The effect of partial sleep deprivation and driving duration on subsequent alertness and performance in car drivers was investigated. Twenty healthy male subjects, between 25 and 55 years of age, free from any sleep disorder, took part in two simulated driving sessions carried out between 2 p.m. and 4 p.m. Before one session, subjects were sleep deprived as they were allowed to sleep only between 3 a.m. and 7 a.m. during the preceding night. Throughout the driving task, the subjects' driving performance, electroencephalogram and Karolinska Sleepiness Scale (KSS) score were recorded. The results revealed that sleep deprivation had an effect on KSS score but not on the (alpha+theta) spectral power, while driving duration had an effect on these two parameters. This effect was also influenced by sleep restriction. Time on driving task alone had a significant effect on driving performance; the sleep restriction having only an effect on one of the performances indices studied: the number of right edge-line crossings. These results are interpreted in terms of the relationship between level of alertness and performance impairment. PMID- 15885248 TI - Radiotelemetry recording of electroencephalogram in piglets during rest. AB - A wireless recording system was developed to study the electroencephalogram (EEG) in unrestrained, male Landrace piglets. Under general anesthesia, ball-tipped silver/silver chloride electrodes for EEG recording were implanted onto the dura matter of the parietal and frontal cortex of the piglets. A pair of miniature preamplifiers and transmitters was then mounted on the surface of the skull. To examine whether other bioelectrical activities interfere with the EEG measurements, an electrocardiogram (ECG) or electromyogram (EMG) of the neck was simultaneously recorded with the EEG. Next, wire electrodes for recording movement of the eyelid were implanted with EEG electrodes, and EEG and eyelid movements were simultaneously measured. Power spectral analysis using a Fast Fourier Transformation (FFT) algorithm indicates that EEG was successfully recorded in unrestrained piglets, at rest, during the daytime in the absence of interference from ECG, EMG or eyelid movements. These data indicate the feasibility of using our radiotelemetry system for measurement of EEG under these conditions. PMID- 15885249 TI - Long-term treadmill exercise overcomes ischemia-induced apoptotic neuronal cell death in gerbils. AB - It has been suggested that exercise may ameliorate neurologic impairment by impeding neuronal loss following various brain insults. In the present study, the effect of long-term treadmill exercise on short-term memory and apoptotic neuronal cell death in the hippocampus following transient global ischemia in gerbils was investigated. A step-down inhibitory avoidance task, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and immunohistochemistry for caspase-3 were used for this study. Ischemia was induced by occlusion of both the common carotid arteries of gerbils for 5 min. Gerbils in the exercise groups were forced to run on a treadmill for 30 min once a day for 4 consecutive weeks. The present results reveal that treadmill exercise for 4 weeks improved short-term memory by suppressing the ischemia-induced apoptotic neuronal cell death in the hippocampus. Here in this study, we show that long-term treadmill exercise for 4 weeks overcomes the ischemia-induced apoptotic neuronal cell death and thus facilitates the recovery of short-term memory impairment induced by ischemic cerebral injury. PMID- 15885250 TI - Seasonal energetics of the Hottentot golden mole at 1500 m altitude. AB - Winter is an energetically stressful period for small mammals as increasing demands for thermoregulation are often coupled with shortages of food supply. In sub-tropical savannah, Hottentot golden moles (Ambysomus hottentottus longiceps) forage throughout the year and for long periods of each day. This may enable them to acquire sufficient resources from an insectivorous prey base that is both widely dispersed and energetically costly to obtain. However, they also inhabit much cooler regions; how their energy budgets are managed in these areas is unknown. We measured the daily energy expenditure (DEE), resting metabolic rate (RMR) and water turnover (WTO) of free-living golden moles during both winter and summer at high altitude (1500 m). We used measurements of deuterium dilution to estimate body fat during these two periods. DEE, WTO and body mass did not differ significantly between seasons. However, RMR values were higher during the winter than the summer and, in the latter case were also lower than allometric predictions. Body fat was also higher during the winter. Calculations show that during the winter they may restrict activity to shorter, more intense periods. This, together with an increase in thermal insulation, might enable them to survive the cold. PMID- 15885251 TI - Early postnatal ethanol administration does not affect prepulse inhibition in rats. AB - Human prenatal ethanol exposure is associated with relatively widespread cognitive deficits but it is unclear whether general deficits in responsivity to sensory stimuli contribute to or underlie the deficits in later or more complex stages of information processing. The present experiment assessed the effects of early postnatal ethanol administration in rats on prepulse inhibition, with animals tested in adolescence (postnatal day (PD) 35) and early adulthood (PD 70). Animals were assigned to receive ethanol (5.25 g/kg) via intragastric intubation on PD 4-9, sham-intubation, or to a naive control group. Pre-exposure to ethanol did not differentially affect the magnitude of the response to the startle stimulus alone nor did it affect the percent inhibition of the startle response on trials with a prepulse stimulus. Male rats exhibited a greater percent inhibition than female rats on PD 35 at all interstimulus intervals (ISIs) except the shortest, 4 ms. Female rats exhibited a greater percent inhibition than male rats at all ISIs on PD 70. Collectively, these data demonstrate that cognitive deficits associated with early exposure to ethanol may not be attributable to deficits in sensorimotor gating, at least to the extent this construct is measured by prepulse inhibition. PMID- 15885252 TI - Rapid learning and flexible memory in "habit" tasks in rats trained with brain stimulation reward. AB - Two groups of rats, one rewarded with sweetened food and the other rewarded with medial forebrain bundle (MFB) stimulation, were trained to home in on and dig for a buried object coated with a target odor. After each group had 15 training trials, MFB rats searched with greater accuracy and speed than food-rewarded rats. MFB rats were subsequently tested (1) after 6 weeks with no additional practice; (2) with food or non-food distractor odors, and (3) with major spatial alterations to the search environment, and in all cases searched with the same high accuracy, short search time, and low level of distractibility as in baseline. These results suggest that the high motivation provided by MFB reward engenders rapidly formed, long-lasting, and surprisingly flexibly deployable "habit" memories. PMID- 15885253 TI - Behavioral correlates of olfactory and trigeminal gaseous stimulation in chickens, Gallus domesticus. AB - Despite growing evidence of avian olfactory abilities, there are few reports of behavioral correlates of chemosensory stimulation in birds. The present study aimed to determine how the behavioral responses of hens to selected gases might be affected by input from different chemosensory systems. We also hoped to relate electrophysiological thresholds previously measured in our laboratory to behavioral evidence of perception. Immediate behavioral responses to ascending series of short (7 s) pulses of olfactory (hydrogen sulphide), trigeminal (carbon dioxide) and combining (ammonia) gaseous stimulants were measured in 12 partially restrained adult hens (Gallus domesticus) using a purpose built gas delivery system. The concentration ranges applied matched those used in our previous electrophysiological studies of olfactory bulb responses. Consistent and specific behavioral responses were observed in response to stimulation with each gas. While significant increases in mandibulation (a distinctive rapid bout of bill movements), interruption of ongoing behavior, orientation to the end of the stimulus, and struggling were seen during stimulation with all three gases, orientation towards the stimulus occurred only in response to the olfactory stimulants, ammonia and hydrogen sulphide. The only gas to elicit significant avoidance was hydrogen sulphide, while gasping and headshaking were elevated in response to stimulation with carbon dioxide. Approximate threshold values for some types of behavior related reasonably well to receptor thresholds determined electrophysiologically, but perception may have occurred at lower concentrations without overt behavioral consequences. The diversity of the behavioral responses observed supports the notion that the gases selectively stimulated different sensory pathways. PMID- 15885254 TI - Concurrent measures of feeding and locomotion in rats. AB - Psychostimulants including amphetamine and cocaine induce locomotion and stereotypy and suppress eating. Studies of the biobehavioral actions of psychostimulants commonly focus on locomotion and less commonly on feeding, and only rarely are these measures considered in conjunction within the same animal. Inasmuch as hyperactivity induced by a psychostimulant may compete with other motor behaviors, including eating, it would be important to concurrently assess changes in eating and locomotion after psychostimulant treatment. The present paper describes a modification of an automated activity chamber in which minute by-minute recordings of food consumption are gathered in parallel with an assessment of locomotion. The present experiment illustrates the method by characterizing the temporal changes in locomotion and eating produced by administration of hypophagic doses of nicotine tartrate (0.28 mg/kg (as the base), IP) or cocaine hydrochloride (7.5 mg/kg, IP). At these doses, nicotine suppressed eating and locomotion, whereas cocaine suppressed eating, but facilitated forward locomotion. These outcomes support the viability of this apparatus and the concurrent method for the dissociation of feeding and locomotion. PMID- 15885255 TI - Cholecystokinin and GABA interaction in the dorsal hippocampus of rats in the elevated plus-maze test of anxiety. AB - In the present study, we have investigated the effects and interaction of CCK and GABAergic systems in the dorsal hippocampus of rats using the elevated plus-maze test of anxiety. Bilateral injection of different doses of CCK(8s) (0.01, 0.05 and 0.1 microg/rat) into the dorsal hippocampus (intra-CA1) decreased percentage of open arm time (%OAT) and open arm entries (%OAE) that are representative of anxiogenic-like behavior. The bilateral injection of three doses of LY225910, a selective CCK2 receptor antagonist (0.01, 0.1 and 0.5 microg/rat) produced significant anxiolytic behavior. Although muscimol (GABA(A+)) (0.1, 0.5 and 1 microg/rat, intra-CA1) produced dose dependent increase in %OAT and a slight increase in %OAE, bicuculline (GABA(A-)), (1, 2 and 4 microg/rat, intra-CA1) failed to change the anxiety profile. Both muscimol (0.1 microg/rat) and bicuculline (1 microg/rat), when co-administered with LY225910, reversed the effect of latter drug on anxiety but when co-administered with CCK8s (0.05 microg/rat) showed no effect on anxiety profile. In conclusion, it seems that both CCK and GABAergic systems not only play a part in the modulation of anxiety in the dorsal hippocampus of rats but also have demonstrated a complex interaction as well. PMID- 15885256 TI - Decrease in nasal temperature of rhesus monkeys (Macaca mulatta) in negative emotional state. AB - We established an infrared thermographic system for the detection of emotion related temperature changes in rhesus monkeys (Macaca mulatta). We continuously measured temperatures of various facial regions of four rhesus monkeys during the presentation of a potentially 'threatening' person, i.e., a person in a laboratory coat with a catching net, who entered the experimental room and approached the monkeys. The temperatures were also measured before and after the presentation of the 'stimulation period.' The temperature of the nasal region decreased significantly within 10-30 s, and continued to decrease throughout the stimulation period. During this period, the monkeys frequently expressed silent bared-teeth face, staring open-mouth face, and lip-smacking, all of which were expressions of a negative emotion. Assuming that the monkeys experience the negative emotion when viewing the potentially threatening stimulus, we conclude that the decrease in nasal skin temperature is relevant to the alteration of the emotional state. The present findings suggest that nasal temperature can be a reliable and accurate indicator of a change from neutral to negative in emotional state of non-human primates. PMID- 15885257 TI - The phasing of circadian rhythms in mice kept under normal or short photoperiods. AB - Mammals represent a multi-oscillatory system. Not only different rhythmic function but also a certain rhythmic function may be controlled by several oscillators. Also, the LD cycle, though being the main zeitgeber, may have different effects on different rhythmic functions. The aim of the present study was to investigate putative changes of internal phase relationships between different rhythms and their phase relationship with respect to the light-dark cycle following a gradual lengthening of the dark time. The investigations were carried out on adult female mice (HaZ:ICR). Animals were initially kept in a 12:12 h LD cycle. The dark time was then lengthened once a week by delaying the time of lights-on by 1 h until an L/D=6:18 h was reached. The motor activity was recorded continuously. The daily profiles of food intake, liver glycogen and melatonin in the serum and the pineal gland were estimated as transverse studies under L/D=12:12 h and L/D=6:18 h. Under short-day conditions (L/D=6:18 h), the evening onset of the main maximum of motor activity and food intake was delayed in relation to lights-off and the onset of the secondary maximum in the morning was advanced in relation to lights-on. Similar phase changes were found with respect to the onset of glycogen synthesis and its breakdown. In the case of melatonin, the rise and the fall occurred earlier or later, respectively, and the phase differences with respect to lights-off and lights-on became smaller. If the mid-point between the evening and morning onsets of activity and food intake and between the rises and falls of glycogen and melatonin concentrations are considered, then the changes in all rhythmic functions were similar. Consequently, the phase relationships between different rhythmic functions and with respect to the light-dark cycle, and thus the internal and the external temporal order, were preserved under different light-dark ratios. Also, the results are in good accordance with a modified two-oscillator model where lights off triggers advances of an evening oscillator and lights-on delays a morning oscillator. PMID- 15885258 TI - Pup exposure differentially enhances foraging ability in primiparous and nulliparous rats. AB - The role of maternal experience (i.e., pregnancy and pup exposure) on rats' performance in a foraging task was assessed. Primiparous (P) and nulliparous (N) animals were either exposed to pups for 21 days (+) or received no pup exposure ( ). Following habituation trials, all animals were tested in spatial and cued versions of the dry land maze (DLM) for three days (three trials per day). In the spatial DLM, the presence of pups decreased latencies in both N and P groups in Trial 5 and P+ rats exhibited shorter latencies to baited food wells than P- animals on Trial 6. In the subsequent probe trial, P+ animals spent significantly more time in proximity to the previously baited well than P- rats. Pups enhanced performance of both P+ and N+ groups in trial 6 of the cued test. Thus, in the spatial task, the individual components of the maternal experience (e.g., pregnancy, parturition, lactation, and pup exposure) converge to produce behavioral modifications in the DLM spatial and probe tasks that enable the female to care for her offspring, in this case, by enhancing foraging behavior. Further, in one trial of each version of the task, pup exposure enhanced performance in N animals suggesting that, in isolation, pup exposure may be a more important influence on ancillary maternal behavior than the pregnancy itself. PMID- 15885259 TI - Behavioral conditioning with interferon beta-1a in humans. AB - Behavioral conditioning is one of the most impressive demonstrations of brain immune system interaction. Numerous animal studies have demonstrated behavioral conditioned effects on immune functions, however, human studies are rare. We investigated whether it is possible to behaviorally condition the acute response to interferon (IFN)beta-1a. In a double-blind placebo-controlled study, 30 healthy subjects received a single injection of IFN(beta)-1a (6MIU of REBIF, Serono International) (unconditioned stimulus, UCS) together with a novel drink (conditioned stimulus, CS). Blood was drawn at baseline, 4, 8, and 24 h after drug administration. Within the first 8 h peripheral granulocytes significantly increased, while monocytes, lymphocytes, T-, B- and natural killer (NK) cell numbers were significantly reduced. In parallel, body temperature, heart rate, norepinephrine and interleukin (IL)-6 plasma levels were heightened within 8 h after injection. 8 days later, all previously IFN(beta)-treated subjects received a subcutaneous placebo (NaCl) injection, but only 15 subjects were re-exposed to the CS (experimental group), while a control group (N=15) drank water and an additional group of subjects (n=8) remained untreated (untreated group). Blood sampling was performed at baseline and at 4, 8, and 24 h. Re-exposition to the CS did not elicit conditioned responses in the experimental group. Moreover, no differences were observed between groups. These data provide negative findings regarding behavioral conditioning of cytokine effects in humans employing a one trial learning paradigm. PMID- 15885260 TI - Microarray analysis of altered gene expression in murine fibroblasts transformed by nickel(II) to nickel(II)-resistant malignant phenotype. AB - B200 cells are Ni(II)-transformed mouse BALB/c-3T3 fibroblasts displaying a malignant phenotype and increased resistance to Ni(II) toxicity. In an attempt to find genes whose expression has been altered by the transformation, the Atlas Mouse Stress/Toxicology cDNA Expression Array (Clontech Laboratories, Inc., Palo Alto, CA) was used to analyze the levels of gene expression in both parental and Ni(II)-transformed cells. Comparison of the results revealed a significant up- or downregulation of the expression of 62 of the 588 genes present in the array (approximately 10.5%) in B200 cells. These genes were assigned to different functional groups, including transcription factors and oncogenes (9/14; fractions in parentheses denote the number of up-regulated versus the total number of genes assigned to this group), stress and DNA damage response genes (11/12), growth factors and hormone receptors (6/9), metabolism (7/7), cell adhesion (2/7), cell cycle (3/6), apoptosis (3/4), and cell proliferation (2/3). Among those genes, overexpression of beta-catenin and its downstream targets c-myc and cyclin D1, together with upregulated cyclin G, points at the malignant character of B200 cells. While the increased expression of glutathione (GSH) synthetase, glutathione-S-transferase A4 (GSTA4), and glutathione-S-transferase theta (GSTT), together with high level of several genes responding to oxidative stress, suggests the enforcement of antioxidant defenses in Ni-transformed cells. PMID- 15885261 TI - Functional genomics may allow accurate categorization of the benzimidazole fungicide benomyl: lack of ability to act via steroid-receptor-mediated mechanisms. AB - Although benomyl and its metabolite carbendazim have been shown to adversely affect male reproduction, the mechanisms of action do not appear to involve the endocrine system. However, few studies have been conducted using currently proposed tests specifically focused on endocrine disruption. Here, potential estrogen- and androgen-mediated activity of benomyl was therefore investigated in vitro and in vivo. Benomyl and carbendazim proved negative for agonistic and antagonistic activity in reporter gene assays for the human estrogen receptor alpha and androgen receptor. In uterotrophic and Hershberger assays using Crj:CD(SD)IGS rats, benomyl (100, 300 or 1000 mg/kg/day, p.o., N = 6) did not exert agonistic effects. However, the highest dose decreased uterine weights in the uterotrophic assay, and decreased weights of some androgen-related tissues of castrated rats receiving a testosterone propionate (TP, 0.2 mg/kg) injection in the Hershberger assay; the effects were less severe than those with p,p'-DDE (100 mg/kg/day). When 4 mg/kg/day of TP was injected, decrease of organ weights due to benomyl was attenuated but still observed. Thus, its influence in some tissues was more potent than that of p,p'-DDE. Benomyl had no apparent effects on serum androgen levels. Microarray analysis of the gene expression profile in the ventral prostate of TP-injected castrated rats treated with benomyl indicated clear differences from the patterns observed with p,p'-DDE and flutamide. Taken together, these findings suggest the decreased organ weights observed in vivo to be caused by mechanisms that are not steroid-receptor-mediated, such as interfering with assembly of microtubules by benomyl. The study furthermore suggests that functional genomics may provide a reliable evidence for accurate categorization of test chemicals. PMID- 15885262 TI - Pharmacokinetic and pharmacodynamic interaction for a binary mixture of chlorpyrifos and diazinon in the rat. AB - Chlorpyrifos (CPF) and diazinon (DZN) are two commonly used organophosphorus (OP) insecticides and a potential exists for concurrent exposures. The primary neurotoxic effects from OP pesticide exposures result from the inhibition of acetylcholinesterase (AChE). The pharmacokinetic and pharmacodynamic impact of acute binary exposures of rats to CPF and DZN was evaluated in this study. Rats were orally administered CPF, DZN, or a CPF/DZN mixture (0, 15, 30, or 60 mg/kg) and blood (plasma and RBC), and brain were collected at 0, 3, 6, 12, and 24 h postdosing, urine was also collected at 24 h. Chlorpyrifos, DZN, and their respective metabolites, 3,5,6-trichloro-2-pyridinol (TCP) and 2-isopropyl-4 methyl-6-hydroxypyrimidine (IMHP), were quantified in blood and/or urine and cholinesterase (ChE) inhibition was measured in brain, RBC, and plasma. Coexposure to CPF/DZN at the low dose of 15/15 mg/kg did not alter the pharmacokinetics of CPF, DZN, or their metabolites in blood. A high binary dose of 60/60 mg/kg increased the C(max) and AUC and decreased the clearance for both parent compounds, likely due to competition between CPF and DZN for CYP450 metabolism. At lower doses, most likely to be encountered in occupational or environmental exposures, the pharmacokinetics were linear. A dose-dependent inhibition of ChE was noted in tissues for both the single and coexposures, and the extent of inhibition was plasma > RBC > or = brain. The overall relative potency for ChE inhibition was CPF/DZN > CPF > DZN. A comparison of the ChE response at the low binary dose (15/15 mg/kg), where there were no apparent pharmacokinetic interactions, suggested that the overall ChE response was additive. These experiments represent important data concerning the potential pharmacokinetic and pharmacodynamic interactions for pesticide mixtures and will provide needed insight for assessing the potential cumulative risk associated with occupational or environmental exposures to these insecticides. PMID- 15885263 TI - Azithromycin and erythromycin ameliorate the extent of colonic damage induced by acetic acid in rats. AB - Ulcerative colitis is a common inflammatory bowel disease (IBD) of unknown etiology. Recent studies have revealed the role of some microorganisms in the initiation and perpetuation of IBD. The role of antibiotics in the possible modulation of colon inflammation is still uncertain. In this study, we evaluated the effects of two macrolides, namely azithromycin and erythromycin, at different doses on the extent and severity of ulcerative colitis caused by intracolonic administration of 3% acetic acid in rats. The lesions and the inflammatory response were assessed by histology and measurement of myeloperoxidase (MPO) activity, nitric oxide synthetase (NOS) and tumor necrosis factor alpha (TNFalpha) in colonic tissues. Inflammation following acetic acid instillation was characterized by oedema, diffuse inflammatory cell infiltration and necrosis. Increase in MPO, NOS and TNFalpha was detected in the colonic tissues. Administration of either azithromycin or erythromycin at different dosage (10, 20 and 40 mg/kg orally, daily for 5 consecutive days) significantly (P < 0.05) reduced the colonic damage, MPO and NOS activities as well as TNFalpha level. This reduction was highly significant with azithromycin when given at a dose of 40 mg/kg. It is concluded that azithromycin and erythromycin may have a beneficial therapeutic role in ulcerative colitis. PMID- 15885264 TI - Gene expression profiling in mouse lung following polymeric hexamethylene diisocyanate exposure. AB - Isocyanates are a common cause of occupational lung disease. Hexamethylene diisocyanate (HDI), a component of polyurethane spray paints, can induce respiratory symptoms, inflammation, lung function impairment, and isocyanate asthma. The predominant form of HDI in polyurethane paints is a nonvolatile polyisocyanate known as HDI biuret trimer (HDI-BT). Exposure of mice to aerosolized HDI-BT results in pathological effects, including pulmonary edema, lung inflammation, cellular proliferation, and fibrotic lesions, which occur with distinct time courses following exposure. To identify genes that mediate lung pathology in the distinct temporal phases after exposure, gene expression profiles in HDI-BT-exposed C57BL/6J mouse lungs were analyzed. RNase protection assay (RPA) of genes involved in apoptosis, cell survival, and inflammation revealed increased expression of IkappaBalpha, Fas, Bcl-X(L), TNFalpha, KC, MIP 2, IL-6, and GM-CSF following HDI-BT exposure. Microarray analysis of approximately 10000 genes was performed on lung RNA collected from mice 6, 18, and 90 h after HDI-BT exposure and from unexposed mice. Classes of genes whose expression was increased 6 h after exposure included those involved in stress responses (particularly oxidative stress and thiol redox balance), growth arrest, apoptosis, signal transduction, and inflammation. Types of genes whose expression was increased at 18 h included proteinases, anti-proteinases, cytoskeletal molecules, and inflammatory mediators. Transcripts increased at 90 h included extracellular matrix components, transcription factors, inflammatory mediators, and cell cycle regulators. This characterization of the gene expression profile in lungs exposed to HDI-BT will provide a basis for investigating injury and repair pathways that are operative during isocyanate-induced lung disease. PMID- 15885265 TI - Pharmacokinetic and pharmacodynamic interactions of morin and cyclosporin. AB - Morin is a flavonoid present in mulberry and herbs. We have reported that morin exerted anti-inflammatory activity on the activated macrophages. Cyclosporin (CsA) is a potent immunosuppressive agent with narrow therapeutic range, which is widely used for the treatments of autoimmune diseases and transplantation rejection. This study aimed to measure the effects of morin on the disposition of CsA in lymphoid and non-lymphoid tissues, and on the functions of immune cells in mice. CsA (Neoral, 10 mg/kg) was orally administered with and without a concomitant dose of morin (0, 50, 100, 200 mg/kg) to mice once daily for 2 weeks. CsA concentrations in blood, liver, kidney, and spleen were determined by a specific monoclonal fluorescence polarization immunoassay. The decreased levels of CsA in tissues were found well correlated to increased doses of morin. The coadministration of 200 mg/kg morin significantly decreased CsA in blood, liver, kidney, and spleen by 33%, 17%, 38%, and 45%, respectively. On the other hand, coadministration of morin decreased dramatically the nitric oxide production by the activated macrophages when compared to CsA treatment alone. Moreover, morin maintained the level of CsA-suppressed T helper 1 (Th1) type cytokine, although the CsA concentration in spleen was markedly reduced. In conclusion, morin coadministration profoundly reduced CsA concentration but did not significantly alter the CsA-suppressed Th1 immune response in mice. PMID- 15885266 TI - An interspecies comparison of mercury inhibition on muscarinic acetylcholine receptor binding in the cerebral cortex and cerebellum. AB - Mercury (Hg) is a ubiquitous pollutant that can disrupt neurochemical signaling pathways in mammals. It is well documented that inorganic Hg (HgCl(2)) and methyl Hg (MeHg) can inhibit the binding of radioligands to the muscarinic acetylcholine (mACh) receptor in rat brains. However, little is known concerning this relationship in specific anatomical regions of the brain or in other species, including humans. The purpose of this study was to explore the inhibitory effects of HgCl(2) and MeHg on [(3)H]-quinuclidinyl benzilate ([(3)H]-QNB) binding to the mACh receptor in the cerebellum and cerebral cortex regions from human, rat, mouse, mink, and river otter brain tissues. Saturation binding curves were obtained from each sample to calculate receptor density (B(max)) and ligand affinity (K(d)). Subsequently, samples were exposed to HgCl(2) or MeHg to derive IC50 values and inhibition constants (K(i)). Results demonstrate that HgCl(2) is a more potent inhibitor of mACh receptor binding than MeHg, and the receptors in the cerebellum are more sensitive to Hg-mediated mACh receptor inhibition than those in the cerebral cortex. Species sensitivities, irrespective of Hg type and brain region, can be ranked from most to least sensitive: river otter > rat > mink > mouse > humans. In summary, our data demonstrate that Hg can inhibit the binding [(3)H]-QNB to the mACh receptor in a range of mammalian species. This comparative study provides data on interspecies differences and a framework for interpreting results from human, murine, and wildlife studies. PMID- 15885267 TI - Volatile organic compounds inhibit human and rat neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes. AB - The relative sensitivity of rats and humans to volatile organic compounds (VOCs) such as toluene (TOL) and perchloroethylene (PERC) is unknown and adds to uncertainty in assessing risks for human exposures to VOCs. Recent studies have suggested that ion channels, including nicotinic acetylcholine receptors (nAChRs), are targets of TOL effects. However, studies comparing TOL effects on human and rat ligand-gated ion channels have not been conducted. To examine potential toxicodynamic differences between these species, the sensitivity of human and rat nAChRs to TOL was assessed. Since PERC has similar effects, in vivo, to TOL, effects of PERC on nAChR function were also examined. Two-electrode voltage-clamp techniques were utilized to measure acetylcholine-induced currents in neuronal nAChRs (alpha4beta2, alpha3beta2, and alpha7) expressed in Xenopus oocytes. PERC (0.065 mM) inhibited alpha7 nAChR currents by 60.1 +/- 4.0% (human, n = 7) and 40 +/- 3.5% (rat, n = 5), and inhibited alpha4beta2 nAChR currents by 42.0 +/- 5.2% (human, n = 6) and 52.2 +/- 5.5% (rat, n = 8). Likewise, alpha3beta2 nAChRs were significantly inhibited by 62.2 +/- 3.8% (human, n = 7) and 62.4 +/- 4.3% (rat, n = 8) in the presence of 0.065 mM PERC. TOL also inhibited both rat and human alpha7, alpha4beta2, and alpha3beta2 nAChRs. Statistical analysis indicated that although there was not a species (human vs. rat) difference with PERC (0.0015-0.065 mM) or TOL (0.03-0.9 mM) inhibition of alpha7, alpha4beta2, or alpha3beta2 nAChRs, all receptor types were more sensitive to PERC than TOL. These results demonstrate that human and rat nACh receptors represent a sensitive target for VOCs. This toxicodynamic information will help decrease the uncertainty associated with animal to human extrapolations in the risk assessment of VOCs. PMID- 15885268 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin-induced change in intestinal function and pathology: evidence for the involvement of arylhydrocarbon receptor-mediated alteration of glucose transportation. AB - Although numerous studies have been performed to clarify the mechanism(s) underlying the toxicological responses induced by dioxins, their effect on the intestine is less well understood. To address this issue, we examined the effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the pathology and function of the intestine in arylhydrocarbon receptor (AhR)-sensitive (C57BL/6J) and -less sensitive (DBA/2J) mice. A single oral administration of TCDD (100 mug/kg) to C57BL/6J mice produced changes in villous structure and nuclear/cytoplasm ratio in the epithelial cells of the intestine. Furthermore, in an oral glucose tolerance test, the serum glucose level was significantly increased in the C57BL/6J mouse but not in the DBA/2J mouse by TCDD treatment. In agreement with this, the expression of intestinal mRNAs coding sodium-glucose co-transporter 1 (SGLT1) and glucose transporter type 2 were increased only in C57BL/6J mice by TCDD. The increase in the former transporter was also confirmed from its protein level. The glucose level in the intestinal contents is thought to be one of the factors contributing to SGLT1 induction. Concerning with this, the intestinal activity of sucrase and lactase was significantly increased only in C57BL/6J mice by TCDD. These results suggest that while TCDD produces initial damage to the intestinal epithelium, the tissues induce SGLT1 to facilitate the absorption of glucose, which is expected, at least partially, to combat the wasting syndrome induced by TCDD. The data provided here also suggest that AhR is involved in the mechanism of SGLT1 induction. PMID- 15885269 TI - In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats disrupts brain sexual differentiation. AB - The effects of in utero and lactational exposure of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) on brain sexual differentiation were investigated. TCDD was orally administered to pregnant Holtzman rats on gestation day (GD) 15, and the activity of brain aromatase, a key enzyme for sexual differentiation, was measured in offspring on postnatal day (PND) 2. Changes in sexual dimorphisms of saccharin preference and the volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA) were examined in adult offspring. In controls, litter means of brain aromatase activity were higher in males than in females. In utero exposure to 200 ng/kg TCDD significantly decreased the sex ratio of aromatase activity (male/female) on PND 2. Offspring were weaned on PND28 and the saccharin test was started on PND84. In controls, saccharin (0.25%) intake (g/kg body weight) was significantly higher in female offspring than in males. In utero exposure to 200 ng/kg TCDD significantly increased saccharin intake in male offspring compared with control males, whereas 800 ng/kg TCDD had no effect. Neither dose of TCDD influenced saccharin intake of female offspring. In controls, SDN-POA volume was significantly greater in males than in females at 14 weeks of age. Exposure to 200 ng/kg TCDD significantly decreased SDN-POA volume in males, whereas 800 ng/kg TCDD had no effect. Neither doses of TCDD influenced the SDN-POA volume in female offspring. These results suggest that in utero and lactational TCDD exposure dose dependently induces demasculinization in male offspring by inhibiting brain aromatase activity in the hypothalamus-preoptic area during central nervous system development. PMID- 15885271 TI - The current role of chemotherapy in metastatic hormone-refractory prostate cancer. AB - Since the publication of the Southwest Oncology Group (SWOG) 99-16 and TAX 327 studies, which demonstrated a survival benefit for docetaxel-based therapy, clinicians for the first time have a therapy to offer men with metastatic prostate cancer that is not merely palliative in its effects. Phase 2 and phase 3 trials are now building on the findings of SWOG 99-16 and TAX 327 by evaluating the potential of combination taxane-based therapies, such as docetaxel plus high dose calcitriol, docetaxel-estramustine-bevacizumab, and docetaxel-thalidomide. The optimal timing of docetaxel-based chemotherapy is still unknown, as there are no prospective clinical trial data to indicate whether earlier treatment (eg, at the time of prostate-specific antigen failure) is more or less effective than later treatment (eg, in metastatic and/or symptomatic disease). PMID- 15885273 TI - Future therapies in hormone-refractory prostate cancer. AB - Hormone-refractory prostate cancer (HRPC) remains true to its name: it is largely refractory to attempts to delay its progression. Although the number of men presenting with metastatic prostate cancer has decreased significantly over the last several years, the death rate for those men is essentially unchanged. To alter the currently inevitable progression of HRPC to death, new targets and new therapies are needed. This article reviews investigational therapies directed against standard targets (eg, the hypothalamic-pituitary-gonadal axis) as well as novel targets (eg, the endothelin axis). PMID- 15885275 TI - Multidisciplinary management of advanced prostate cancer: changing perspectives on referring patients and enhancing collaboration between oncologists and urologists in clinical trials. AB - Prostate cancer traditionally was managed by monotherapy, and treatment was almost exclusively provided by the urologic surgeon. As ongoing clinical trials in advanced disease identify novel agents or combination therapies with good tolerability and potential survival benefits, multidisciplinary management may offer optimal care for men with high-risk prostate cancer, as it does for women with breast cancer. Patients with high-risk recurrent or metastatic disease should be informed of clinical trials for which they may be eligible. PMID- 15885277 TI - National cooperative group trials. AB - Now is the time to answer critical questions in the management of prostate cancer through enrollment in high-priority national phase 3 clinical trials. Studies of systemic therapy before and after local therapy are vitally important to determine whether early systemic interventions might delay cancer progression. Chemotherapy must be studied in earlier disease progression. Novel targeted therapies, such as vaccine therapy, angiogenesis inhibitors, and newer microtubule targeting agents, should be studied in all stages of progression. Despite these needs and the availability of systemic therapies, accrual to trials has been lagging. This article reviews the national cooperative group clinical trials, whether currently open or planned, and suggests practical strategies to improve accrual. PMID- 15885279 TI - Vitamin D and fractures: quo vadis? PMID- 15885280 TI - Smoking cessation, weight gain, and lung function. PMID- 15885281 TI - Feeder-free derivation of human embryonic stem-cell lines. PMID- 15885282 TI - Live organ-donation for islet transplantation. PMID- 15885283 TI - Maternal hypothyroxinaemia during (early) gestation. PMID- 15885284 TI - Lymphogranuloma venereum in the industrialised world. PMID- 15885287 TI - Unethical clinical trials in Thailand: a community response. PMID- 15885288 TI - US exceptionalism and research ethics. PMID- 15885289 TI - US exceptionalism and research ethics. PMID- 15885291 TI - Selective head cooling after neonatal encephalopathy. PMID- 15885292 TI - Selective head cooling after neonatal encephalopathy. PMID- 15885294 TI - Oral vitamin D3 and calcium for secondary prevention of low-trauma fractures in elderly people (Randomised Evaluation of Calcium Or vitamin D, RECORD): a randomised placebo-controlled trial. AB - BACKGROUND: Elderly people who have a fracture are at high risk of another. Vitamin D and calcium supplements are often recommended for fracture prevention. We aimed to assess whether vitamin D3 and calcium, either alone or in combination, were effective in prevention of secondary fractures. METHODS: In a factorial-design trial, 5292 people aged 70 years or older (4481 [85%] of whom were women) who were mobile before developing a low-trauma fracture were randomly assigned 800 IU daily oral vitamin D3, 1000 mg calcium, oral vitamin D3 (800 IU per day) combined with calcium (1000 mg per day), or placebo. Participants who were recruited in 21 UK hospitals were followed up for between 24 months and 62 months. Analysis was by intention-to-treat and the primary outcome was new low energy fractures. FINDINGS: 698 (13%) of 5292 participants had a new low-trauma fracture, 183 (26%) of which were of the hip. The incidence of new, low-trauma fractures did not differ significantly between participants allocated calcium and those who were not (331 [12.6%] of 2617 vs 367 [13.7%] of 2675; hazard ratio (HR) 0.94 [95% CI 0.81-1.09]); between participants allocated vitamin D3 and those who were not (353 [13.3%] of 2649 vs 345 [13.1%] of 2643; 1.02 [0.88-1.19]); or between those allocated combination treatment and those assigned placebo (165 [12.6%] of 1306 vs 179 [13.4%] of 1332; HR for interaction term 1.01 [0.75 1.36]). The groups did not differ in the incidence of all-new fractures, fractures confirmed by radiography, hip fractures, death, number of falls, or quality of life. By 24 months, 2886 (54.5%) of 5292 were still taking tablets, 451 (8.5%) had died, 58 (1.1%) had withdrawn, and 1897 (35.8%) had stopped taking tablets but were still providing data for at least the main outcomes. Compliance with tablets containing calcium was significantly lower (difference: 9.4% [95% CI 6.6-12.2]), partly because of gastrointestinal symptoms. However, potentially serious adverse events were rare and did not differ between groups. INTERPRETATION: The findings do not support routine oral supplementation with calcium and vitamin D3, either alone or in combination, for the prevention of further fractures in previously mobile elderly people. PMID- 15885295 TI - Smoking cessation, lung function, and weight gain: a follow-up study. AB - BACKGROUND: Only one population-based study in one country has reported effects of smoking cessation and weight change on lung function, and none has reported the net effect. We estimated the net benefit of smoking cessation, and the independent effects of smoking and weight change on change in ventilatory lung function in the international European Community Respiratory Health Survey. METHODS: 6654 participants in 27 centres had lung function measured in 1991-93, when aged 20-44 years, and in 1998-2002. Smoking information was obtained from detailed questionnaires. Changes in lung function were analysed by change in smoking and weight, adjusted for age and height, in men and women separately and together with interaction terms. FINDINGS: Compared with those who had never smoked, decline in FEV1 was lower in male sustained quitters (mean difference 5.4 mL per year, 95% CI 1.7 to 9.1) and those who quit between surveys (2.5 mL, -1.9 to 7.0), and greater in smokers (-4.8 mL, -7.9 to -1.6). In women, estimates were 1.3 mL per year (-1.5 to 4.1), 2.8 mL (-0.8 to 6.3) and -5.1 mL (-7.5 to -2.8), respectively. These sex differences were not significant. FEV1 changed by -11.5 mL (-13.3 to -9.6) per kg weight gained in men, and by -3.7 mL per kg (-5.0 to 2.5) in women, which diminished the benefit of quitting by 38% in men, and by 17% in women. INTERPRETATION: Smoking cessation is beneficial for lung function, but maximum benefit needs control of weight gain, especially in men. PMID- 15885296 TI - Human embryonic stem cells derived without feeder cells. AB - BACKGROUND: Human embryonic stem cells are likely to play an important role in the future of regenerative medicine. However, exposure of existing human embryonic stem-cell lines to live animal cells and serum risks contamination with pathogens that could lead to human health risks. We aimed to derive an embryonic stem-cell line without exposure to cells or serum. METHODS: Frozen cleavage-stage embryos were thawed and cultured to the blastocyst stage. Inner cell masses were isolated by immunosurgery and plated onto extracellular-matrix-coated plates that can be easily sterilised. Six established human embryonic stem-cell lines were also maintained with this serum and feeder free culture system. FINDINGS: A new stem-cell line was derived from human embryos under completely cell and serum free conditions. The cells maintained normal karyotype and markers of pluripotency, including octamer binding protein 4 (Oct-4), stage-specific embryonic antigen (SSEA)-3, SSEA-4, tumour-rejection antigen (TRA)-1-60, TRA-1 81, and alkaline phosphatase. After more than 6 months of undifferentiated proliferation, these cells retained the potential to form derivatives of all three embryonic germ layers both in vitro and in teratomas. These properties were also successfully maintained (for more than 30 passages) with the established stem-cell lines. INTERPRETATION: This system eliminates exposure of human embryonic stem cells and their progeny to animal and human feeder layers, and thus the risk of contamination with pathogenic agents capable of transmitting diseases to patients. PMID- 15885297 TI - Combined therapy with somatostatin analogues and weekly pegvisomant in active acromegaly. AB - Pegvisomant monotherapy once daily returns concentrations of insulin-like growth factor I (IGF-I) to normal in most patients with acromegaly, but is very costly. In a 42-week dose-finding study, we assessed the efficacy of the combination of long-acting somatostatin analogues once monthly and pegvisomant once weekly in 26 patients with active acromegaly. Dose of pegvisomant was increased until IGF-I concentration became normal or until a weekly dose of 80 mg was reached. IGF-I reached normal concentrations in 18 of 19 (95%) patients who completed 42 weeks of treatment, with a median weekly dose of 60 mg pegvisomant (range 40-80). No signs of pituitary tumour growth were noted, but mild increases in liver enzymes were observed in ten patients (38%). This combined treatment is effective, might increase compliance, and could greatly reduce the costs of medical treatment for acromegaly in some patients. PMID- 15885298 TI - Aplastic anaemia. AB - Aplastic anaemia is a rare haemopoietic stem-cell disorder that results in pancytopenia and hypocellular bone marrow. Although most cases are acquired, there are unusual inherited forms. The pathophysiology of acquired aplastic anaemia is immune mediated in most cases; autoreactive lymphocytes mediate the destruction of haemopoietic stem cells. Environmental exposures, such as to drugs, viruses, and toxins, are thought to trigger the aberrant immune response in some patients, but most cases are classified as idiopathic. Similarly to other autoimmune diseases, aplastic anaemia has a varied clinical course; some patients have mild symptoms that necessitate little or no therapy, whereas others present with life-threatening pancytopenia representing a medical emergency. Paroxysmal nocturnal haemoglobinuria and myelodysplastic syndrome commonly arise in patients with aplastic anaemia, showing a pathophysiological link between these disorders. Acquired aplastic anaemia can be effectively treated by allogeneic bone-marrow transplantation, immunosuppression (generally antithymocyte globulin and ciclosporin), and high-dose cyclophosphamide. PMID- 15885299 TI - Multiplicity in randomised trials II: subgroup and interim analyses. AB - Subgroup analyses can pose serious multiplicity concerns. By testing enough subgroups, a false-positive result will probably emerge by chance alone. Investigators might undertake many analyses but only report the significant effects, distorting the medical literature. In general, we discourage subgroup analyses. However, if they are necessary, researchers should do statistical tests of interaction, rather than analyse every separate subgroup. Investigators cannot avoid interim analyses when data monitoring is indicated. However, repeatedly testing at every interim raises multiplicity concerns, and not accounting for multiplicity escalates the false-positive error. Statistical stopping methods must be used. The O'Brien-Fleming and Peto group sequential stopping methods are easily implemented and preserve the intended alpha level and power. Both adopt stringent criteria (low nominal p values) during the interim analyses. Implementing a trial under these stopping rules resembles a conventional trial, with the exception that it can be terminated early should a treatment prove greatly superior. Investigators and readers, however, need to grasp that the estimated treatment effects are prone to exaggeration, a random high, with early stopping. PMID- 15885300 TI - A swollen calf. PMID- 15885302 TI - Anti-neuronal antibodies and movement disorders. PMID- 15885303 TI - Microinfusion of antineuronal antibodies into rodent striatum: failure to differentiate between elevated and low titers. AB - An autoimmune-mediated mechanism has been proposed for several pediatric movement disorders. In a three-center (Brown, Yale, and Johns Hopkins) collaborative effort, serum antineuronal antibodies (ANAb) were measured by use of ELISA or immunohistochemical techniques on 35 children (mean age 11.4 years) with Tourette syndrome, attention deficit hyperactivity disorder, and/or obsessive compulsive disorder. Eight sera, 4 containing the highest and 4 the lowest levels of ANAb, were identified at each institution. Selected sera (total of 9 with elevated and 7 with low ANAb) were re-encoded and sent to each center for infusion into the ventrolateral striatum of 16 male Sprague-Dawley rats. Animals were observed for behavioral abnormalities for 3 days before the start of infusion, during infusion on days 2-4, and for 2 days after infusion. Combined stereotypy scores increased after antibody infusion, but there was no significant effect based on serum titer (p=0.85). Scores differed among centers, but analyses based on individual institutional data again failed to show an effect based on elevated or low ANAb values (Brown, p=0.95; Yale and Johns Hopkins, p=0.81). Post hoc studies with sham surgery and infusion of phosphate-buffered saline support suggestions of nonspecific behavioral effects unrelated to antibody titer. This report emphasizes that any conclusions about antibody-mediated movement disorders that are based upon results from the rodent infusion model must be considered with caution. PMID- 15885304 TI - A role for brain-derived neurotrophic factor in B cell development. AB - In the present study, we demonstrated a significant reduction of B lymphocytes in the blood, spleen and bone marrow of BDNF deficient mice. The observed developmental block in bone marrow B cell development was linked specifically to the Pre-BII stage. B lymphocytes express the BDNF receptors p75NTR and TrkB(gp95), while no BDNF expression was found. However, a strong BDNF expression was demonstrated in bone marrow stromal cells. An increase of intracellular free calcium [Ca2+]i in B lymphocytes after BDNF application confirms a direct responsiveness of B lymphocytes to BDNF. In conclusion, these results suggest a role of BDNF for normal B lymphocyte development through paracrine effects in the bone marrow. PMID- 15885305 TI - Mechanism of natural killer (NK) cell regulatory role in experimental autoimmune encephalomyelitis. AB - The mechanism of natural killer (NK) cell regulatory role in experimental autoimmune encephalomyelitis (EAE) was studied in SJL/J mice. In vivo experiments showed that NK cell depletion by anti-NK1.1 monoclonal antibody treatment enhanced EAE in mice. To investigate the mechanism, we cultured proteolipid protein (PLP)136-150 peptide-specific, encephalitogenic T cell lines, which were used as the NK cell target. Our results show that NK cells exert a direct cytotoxic effect on autoantigen-specific, encephalitogenic T cells. Furthermore, cytotoxicity to PLP-specific, encephalitogenic T line cells was enhanced by using enriched NK cells as effector cells. However, the cytotoxic effect of NK cells to ovalbumin-specific T line cells and ConA-stimulated T cells could also be detected with a lesser efficiency. Our studies indicate that NK cells play a regulatory role in EAE through killing of syngeneic T cells which include myelin antigen-specific, encephalitogenic T cells, and thus ameliorate EAE. PMID- 15885306 TI - Treatment with chimeric anti-human CD40 antibody suppresses MRI-detectable inflammation and enlargement of pre-existing brain lesions in common marmosets affected by MOG-induced EAE. AB - Common marmosets, a Neotropical monkey species, are protected against clinical and neuropathological consequences of experimentally induced autoimmune encephalomyelitis (EAE) by prophylactic treatment with ch5D12, a humanized antagonist antibody against human CD40. In the current study we have tested whether ch5D12 acts therapeutically against the enlargement and inflammatory activity of existing (brain) white matter lesions using serial magnetic resonance imaging (MRI). The results show in all PBS treated monkeys (n=4) a rapid enlargement of T2 lesions together with an increment of the T2 signal intensity due to inflammatory edema. Treatment with ch5D12 delayed the enlargement of T2 lesions in 2 out of 3 tested monkeys while in 3 out of 3 monkeys the T2 signal increment of lesions was suppressed. In conjunction with previously published data on the clinical benefit of anti-CD40 treatment in the marmoset EAE model, the current findings support antibody-mediated blockade of CD40 interaction with its ligand CD154 as a potential treatment of MS. PMID- 15885307 TI - Suppression of natural killer cell cytotoxicity following chronic electrical stimulation of the ventromedial hypothalamic nucleus in rats. AB - In our previous study we found that chronic electrical stimulation of the lateral hypothalamus (LH) enhances and its lesion suppresses natural killer cell cytotoxicity (NKCC) and a large granular lymphocyte (LGL) number in conscious, freely behaving rats. Since the ventromedial nucleus of the hypothalamus (VMH) is regarded as behaviorally and physiologically opposite to LH, in our present study we investigated whether this antagonism also holds for the immune functions. Chronic electrical VMH stimulation effect on 1) immune parameters: both spleen and blood NKCC (chromium release assay and single-cell agarose assay) and the number of large granular lymphocytes (LGL; a morphological method), and 2) endocrine parameters: immunosuppressive-corticosterone (COR) and testosterone (TST) and immunostimulative-growth hormone (GH) and prolactin (PRL) plasma levels (RIA) was assessed. Twenty-one days of electrical stimulation of VMH caused significant decrease in both spleen and blood NKCC at the population level (chromium release assay) but not at the single cell level (agarose assay) with a simultaneous fall in the LGL number. Rats responding to the VMH stimulation with behavioral inactivation (BIN) showed a significantly lower depression of NKCC and LGL number than those responding with an aversive reaction (AVE). Depression of NKCC coexisted with various hormonal changes: increase of PRL, increase (AVE) or fall (BIN) of COR, decrease of GH (BIN), and increase of TST (VMH-stimulated and VMH-sham). There were significant differences in all measured plasma hormones between BIN and AVE groups. The results obtained indicate that VMH decreases cell mediated immune response, represented by NK cell activity. The immunosuppressive effect is dependent on the behavioral outcome of VMH stimulation (BIN/AVE) rather than tested endocrine variables. Moreover, the present results indicate that the VMH and LH are antagonistically engaged in the regulation of NK cell cytotoxicity. PMID- 15885308 TI - Differential expression of cytokines in the brain and serum during endotoxin tolerance. AB - Using short- and long-course lipopolysaccharide (LPS) treatment regimens to induce endotoxin tolerance in rats, we compared TNF-alpha, IL-1beta, and IL-6 expression in the brain and serum following a LPS challenge. Serum corticosterone was also examined to evaluate the function of the hypothalamic-pituitary-adrenal (HPA) axis. We found that, during endotoxin tolerance, LPS-induced cytokine expression still occurred in the brain even when cytokines in the periphery were no longer induced, and that this differential cytokine expression may be mediated by corticosterone, a stress hormone and an anti-inflammatory agent. PMID- 15885309 TI - Brain macrophage activation in murine cerebral malaria precedes accumulation of leukocytes and CD8+ T cell proliferation. AB - Although the activation of brain macrophages is associated with both human and mouse cerebral malaria (CM) the relative contributions of the heterogeneous populations of brain macrophages to the disease are unknown. In this work, we dissociate for the first time inflammatory monocytes from resident brain macrophages in mice developing CM when infected with Plasmodium berghei. Based on the differential expression of CD45 in brain macrophage cell populations and by using bone-marrow (BM) chimeras reconstituted with GFP cells we clearly distinguish between blood-derived cells and resident brain cells of hematopoietic origin. FACS and histological analysis reveal that although inflammatory monocytes and CD8+ T cells invade the brain during CM, parenchymal macrophages also undergo morphological changes and over express MHC class I and Sca-1. In addition to the leukocyte sequestration in the brain, in situ proliferation contributes to the expansion of CD8+ T cells during CM. Finally, kinetic analysis of brain cells during infection with P. berghei demonstrates that activation of parenchymal macrophages precedes leukocyte sequestration in the brain vasculature. Thus, our results reveal the phenotype of activation of brain macrophages during CM showing that parenchymal brain macrophages are activated before overwhelming brain inflammation. These results further suggest that brain macrophages may contribute to the local proliferation of CD8+ T cells that culminate in death of mice with CM syndrome. PMID- 15885310 TI - Vascular endothelial growth factor is regulated by hypoxic stress via MAPK and HIF-1 alpha in the inner ear. AB - Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis. The iron-chelator desferrioxamine (DFX) increased the expression of hypoxia-inducible factor (HIF)-1alpha in the hair cell line, HEI-OC1. The increased VEGF production by DFX was inhibited by iron. DFX also induced the activation of mitogen activated protein kinase (MAPK) on HEI-OC1. The increased VEGF production by DFX was inhibited by a specific inhibitor of MAPK. In addition, DFX induced the VEGF production and HIF-1alpha stabilization in vivo. These results indicate that VEGF production is regulated via MAPK and HIF-1alpha under hypoxic condition in the inner ear. PMID- 15885311 TI - Evidence for a restricted rather than generalized stimulatory response of skin derived human mast cells to substance P. AB - To resolve the controversy regarding substance P (SP) mediated stimulation of mast cells (MC), we demonstrate that SP triggers histamine release from purified human skin MC (sMC), but contrast to stimulation via FcepsilonRI, does not effect the production of TNF-alpha or IL-8. Conversely, both anti-IgE and SP are suppressive in terms of IL-6. By quantitative RT-PCR, the amount of templates at baseline (per 25 ng total RNA) is 2178 (IL-6), 2,665 (IL-8) and 94 (TNF-alpha), and remains unaltered by SP. Contrast to sMC, LAD2 MC respond to SP with stronger histamine release and robust TNF-alpha production in an only partially neurokinin 1R mediated manner, while histamine release of sMC is chiefly mediated by this receptor. We conclude that human sMC are responsive to SP in a selective manner by eliciting degranulation without the induction of cytokines and that SP triggered cytokine production varies among MC subtypes, likely through differences in signaling mechanisms. PMID- 15885312 TI - Chronic recurrent autoimmune uveitis with progressive photoreceptor damage induced in rats by transfer of IRBP-specific T cells. AB - Recurrent uveitis is a common cause of vision blindness. Using a rat model of chronic recurrent uveitis, we examined the relationship between clinical expression, pathological changes, and the heterogeneity of the disease. Chronic recurrent uveitis was induced by adoptive transfer of interphotoreceptor retinoid binding protein (IRBP)-specific T cells in a total of more than 60 Lewis rats. In about 75% of cases recurrent uveitis was pathologically a chronic and progressive disease. The major pathological changes included the gradual loss of photoreceptor cells. However, disease progression did not always parallel the severity of ocular inflammation and clinical recurrent disease, with about a quarter showing no pathological damage in the eye. PMID- 15885313 TI - Tissue-specific alterations in the glucocorticoid sensitivity of immune cells following repeated social defeat in mice. AB - Endogenous glucocorticoids (GC) play an important role in the termination of the inflammatory response following infection and tissue injury. However, recent findings indicate that stress can impair the anti-inflammatory capacities of these hormones. Lipopolysaccharide (LPS)-stimulated splenocytes of mice that were repeatedly subjected to social disruption (SDR) stress were less sensitive to the immunosuppressive effects of corticosterone (CORT) as demonstrated by an increased production of pro-inflammatory cytokines and enhanced cell survival. Myeloid cells expressing the marker CD11b were shown to play a key role in this process. Here we investigated the role of the bone marrow as a potential source of the GC-insensitive cells. The study revealed that LPS-stimulated bone marrow cells, in the absence of experimental stress, were virtually GC-resistant and retained high levels of cell viability after treatment with CORT. Recurrent exposure to the acute stressor over a period of 2, 4 or 6 days led to an increase in the GC sensitivity of the bone marrow cells. This increase in GC sensitivity was associated with enhanced mRNA expression of granulocyte-macrophage colony stimulating factor (GM-CSF), an increase in the number of myeloid progenitors, and a decrease in the proportion of mature CD11b+ cells. The changes in the cellular composition of the bone marrow were accompanied by an increase in splenic CD11b+ cell numbers. Simultaneous assessment of the GC sensitivity in bone marrow and spleen revealed a significant negative correlation between both tissues suggesting that social stress causes the redistribution of GC-insensitive myeloid cells from the bone marrow to the spleen. PMID- 15885314 TI - Expression of P2X4 receptor by lesional activated microglia during formalin induced inflammatory pain. AB - P2X4 receptor (P2X4R) is an ion channel gated by adenosine 5'-triphosphate. Here we report the presence and the distribution of P2X4R in rat spinal cord by immunohistochemical analysis in an inflammatory pain model. Peripheral inflammation was induced by subcutaneous injection of 4% formalin into the rat hindpaw. Morphology, spatial localization, and activation state of P2X4R+ cells were described at 1, 5, 7, 14, and 28 days after injury. In normal and saline treated control rats, P2X4R was rarely seen. After formalin administration, an increase of P2X4R+ microglia were observed in the spinal cord dorsal horn on the side ipsilateral to the injection, reaching maximal levels by day 7, and then decreasing to normal levels by day 14. This implicates a role of P2X4R in the spinal inflammatory pain process. Furthermore, formalin-induced region-specific increase in activated microglia was confirmed by ED1 and endothelial monocytes activating polypeptide II (EMAP-II) expression. In conclusion, this is the first demonstration that P2X4R is expressed by microglia in the inflammatory pain. PMID- 15885315 TI - Increased expression of CXCR3 and CCR5 on memory CD4+ T-cells migrating into the cerebrospinal fluid of patients with neuroborreliosis: the role of CXCL10 and CXCL11. AB - The aim of this study was to evaluate the contribution of chemokine receptor CXCR3 and the corresponding ligands CXCL10 and CXCL11 to the recruitment of peripheral blood (PB) memory CD4+ T-cells into the cerebrospinal fluid (CSF) of patients with acute neuroborreliosis. Percentages of memory CD45RO+CD4+ T-cells expressing CXCR3 and CCR5 were significantly enriched in the CSF compared to the PB. Concentrations of CXCL10 and CXCL11 in the CSF of neuroborreliosis patients were significantly higher compared with the corresponding serum samples. Our results suggest that CXCL10 and CXCL11 create a chemokine gradient between the CSF and serum and recruite CXCR3-expressing memory CD4+ T-cells into the CSF of neuroborreliosis patients and that CCR5 also plays a role in this process. PMID- 15885316 TI - Development of a rapid autopsy program for studies of brain immunity. AB - Human glia are essential cellular models used for studies of neurodegenerative diseases. Fetal neuroglia are commonly used, as they can be recovered in large quantities and sustained for long periods in culture. However, fetal neuroglia may have limitations in reflecting adult diseases and additionally can pose ethical issues in translating products of abortion for research use. To address these concerns, we developed a rapid autopsy program to procure age- and disease specific neuroglia from adult brain tissues within hours of death. The challenges in developing this initiative, reflecting experiences from 69 autopsies over 4 years, are presented. PMID- 15885317 TI - Gelatinases (MMP-2 and MMP-9) are preferentially expressed by Th1 vs. Th2 cells. AB - T helper subsets, Th1 and Th2, which are associated with different types of immune reactions, are distinguished by their cytokines' profiles and expression of different chemokine receptors, that may differentially influence their migratory capacity. The present study examined the expression of matrix metalloproteinases, (MMP)-2 and MMP-9, by the two CD4+ Th cell subsets and the role of these proteases in their migration. We observed that migration of CD4+ T cells is dependent on the gelatinases, and that the migratory capacity of Th1 is higher than Th2 cells. In addition, Th1 in comparison to Th2 cells, from both human (healthy subjects and multiple sclerosis (MS) patients) as well as from murine origin, secrete higher levels of MMP-2 and MMP-9. These novel findings contribute to the understanding of the physiological Th1 and Th2 immune responses as well as the enhanced Th1 reactivity in immune-mediated diseases, such as MS, in which enhanced levels of MMP-2 and MMP-9 are associated with disease processes. PMID- 15885318 TI - IFNAR1 and IFNAR2 polymorphisms confer susceptibility to multiple sclerosis but not to interferon-beta treatment response. AB - We investigated the role of three polymorphisms in the IFNAR1 (SNPs 18417 and 408) and IFNAR2 (SNP 11876) genes in multiple sclerosis (MS) susceptibility and in the IFNbeta treatment response in a group of 147 patients and 210 controls undergoing interferon therapy during the last 2 years. Only the 18417 and the 11876 SNPs showed an association with disease susceptibility (p=0.001 and 0.035, respectively) although no differential genotype distribution were observed between interferon responders and non-responder MS patients. No alteration of the expression level of IFNAR-1 was observed with respect to the -408 genotypes or to interferon treatment response. These data suggest a role for the IFNAR pathway in susceptibility to MS. PMID- 15885319 TI - Osteopontin gene haplotypes correlate with multiple sclerosis development and progression. AB - Osteopontin (OPN) is an inflammatory cytokine highly expressed in multiple sclerosis (MS) plaques. In a previous work, we showed that four OPN polymorphisms form three haplotypes (A, B, and C) and that homozygotes for haplotype-A display lower OPN levels than non-AA subjects. In this work, we evaluated the distribution of these OPN haplotypes in 425 MS patients and 688 controls. Haplotype-A homozygotes had about 1.5 lower risk of developing MS than non-AA subjects. Clinical analysis of 288 patients showed that AA patients displayed slower switching from a relapsing remitting to a secondary progressive form and milder disease with slower evolution of disability. MS patients displayed increased OPN serum levels, which were partly due to the increased frequency of non-AA subjects. Moreover in AA patients, OPN levels were higher than in AA controls and similar to those found in both non-AA patients and controls, which suggests a role of the activated immune response. These data suggest that OPN genotypes may influence MS development and progression due to their influence on OPN levels. PMID- 15885320 TI - Comparison of ventricular and lumbar cerebrospinal fluid T cells in non inflammatory neurological disorder (NIND) patients. AB - The aim of the present study was to define the cellular composition of ventricular, as compared with lumbar, cerebrospinal fluid (CSF) in patients with non-inflammatory neurological disorders (NIND). We addressed this issue by determining the cellular composition of lumbar CSF from patients with normal pressure hydrocephalus (NPH) who were undergoing lumbar CSF drainage during evaluation for shunting procedures, and evaluating ventricular CSF from a subset of these who underwent subsequent placement of ventriculoperitoneal shunts. We determined the cellular composition of lumbar CSF from 18 patients with NPH, and found that the leukocyte differentials, and relative proportions of CD4+ and CD8+ central memory (TCM), effector memory (TEM) and naive cell (TNaive) populations, were equivalent to those found previously in studies of CSF from patients with NIND. We further evaluated cells in the ventricular CSF of five patients who had previously undergone lumbar drainage. Leukocyte differential counts, as well as CD4+ and CD8+ TCM, TEM, and TNaive proportions, were equivalent in matched ventricular and lumbar CSF samples. These observations support the hypothesis that leukocytes enter the CSF in a selective fashion, at its site of formation in the choroid plexus. The results implicate CSF T cells in the immune surveillance of the central nervous system. PMID- 15885321 TI - Spinal cord injury induces early and persistent lesional P2X4 receptor expression. AB - Following spinal cord injury (SCI), neuropathic, chronic pain is a major cause of disability. Recently, glial P2X4 receptor (P2X4R) has been identified as a major contributor to the development of neuropathic pain after peripheral nerve injury. Here we report analysis of P2X4R expression following rat SCI. Significant lesional accumulation of P2X4R+ cells was detected as early as 24 h after SCI, reaching maximum cell numbers on Day 7. Thereafter cell numbers declined but persisted at significantly elevated, sub-maximal levels (>70%) until 1 month post injury. Double-immunolabeling identified the majority of lesional P2X4R+ cells as activated microglia/macrophages and surviving neurons/neurites. Increase of P2X4R+, beta-APP+ hypertrophic neurites correlated with proximity to the lesion. Further, P2X4R+ cells coexpressed the intracellular regulators of signalling cascades, COX-1 (>20%), COX-2 (>5%), RhoA (>60%) and RhoB (>10%). PMID- 15885322 TI - Evaluation of cerebrospinal fluid uPA, PAI-1, and soluble uPAR levels in HIV infected patients. AB - To evaluate the potential role of the uPAR/uPA/PAI-1 system in HIV-induced blood brain-barrier (BBB) disruption, CSF uPA-dependent plasminogen activation (PdPA) was analyzed by casein zymography, and CSF protein levels of all three molecules were measured by ELISA. CSF uPAR, but not uPA, PAI-1, or PdPA levels was significantly increased in neurologically compromised HIV+ patients. Only individual patients with severe AIDS dementia complex had increased levels of uPA (but not PAI-1) which fell upon initiation of antiretroviral therapy. The levels of all three molecules did not correlate with the CSF to serum albumin ratio suggesting not an important role in HIV-induced BBB disruption. PMID- 15885323 TI - Lymphocyte subsets, TNF alpha and interleukin-4 levels in treated and untreated subacute sclerosing panencephalitis patients. AB - Immunologic studies in relation to clinical status might help to understand the pathogenesis of subacute sclerosing panencephalitis (SSPE) and the effect of treatment. We measured lymphocyte subsets and intracellular TNFalpha and interleukin-4 levels in peripheral blood in SSPE patients. Patients had elevated percentages of CD8+ cells compared to age-matched control children. Rapidly progressive course was associated with increased CD4+ cells. Treatment with interferons and inosiplex altered the percentage of CD3+, CD4+ and CD19+ cells. TNFalpha and interleukin-4 levels had no correlation with course or treatment. The proportions of lymphocyte subsets appear to have a role in the evolution or manifestations of SSPE, if not in the pathogenesis. PMID- 15885324 TI - Can the world's infants with hearing loss wait? PMID- 15885325 TI - Validation of tele-otology to diagnose ear disease in children. AB - OBJECTIVE: To determine if digitised still eardrum images, with a clinical history, and audiometry and tympanometry data provide sufficient information to an ear specialist to make an assessment of a patient. METHODS: 66 children (9 months to 16 years) from remote communities were assessed by an ear specialist by standard otoscopy, using a clinical history, audiometry and tympanometry. Up to five images of each ear were digitised. At a later date, the ear specialist made observations, diagnoses and recommendations for management from the images and clinical data. RESULTS: There was a significant correlation (p<0.01) between image quality and age of the subject. There were significant agreements for the clinically important observations of otorrhea, perforation, retracted tympanic membrane and atrophy of the tympanic membrane (p<0.05). There were significant agreements for the diagnoses of acute otitis media, chronic suppurative otitis media, otitis media with effusion and Eustachian tube dysfunction. The rate of recommendations for review or referral after a tele-otology assessment were between 4 and 16% higher than those in made in the field. The agreements between the various forms of advice or recommendations made in the field to those made by tele-otology were statistically significant (p<0.01). CONCLUSIONS: A tele-otology system that incorporates good quality digitised images of the tympanic membrane, audiological and tympanometric data, and a comprehensive clinical history provides the ear specialist with sufficient information to make a confident diagnose of existing middle ear disease, and provide management advice to the patients' primary care provider. PMID- 15885326 TI - Does Helicobacter pylori have role in development of otitis media with effusion? AB - OBJECTIVE: Recently, it was suggested that tonsil and adenoid tissues may act as a reservoir for Helicobacter pylori (HP). A connection between chronic tubotympanal disorders and gastroesophageal reflux is well recognized, but the mechanism underlying this relationship is unclear. In this study, we investigated possible presence of HP in adenoid tissue and middle ear effusions in patients with chronic otitis media with effusion (OME) and we compared the data with the results of the children who had adenoid hypertrophy without OME. METHODS: The study was comprised of 38 consecutive children with adenoid hypertrophy and/or chronic OME. The patients were divided into two groups. The first group included 18 subjects having OME+adenoid hypertrophy and the second group included 20 subjects having solely adenoid hypertrophy. Each patient underwent the appropriate surgical procedure; myringotomy, placement of tympanostomy tubes and/or adenoidectomy. After myringotomy, the middle ear effusions were collected in a suction and collection device and a core biopsy specimen was taken from each adenoid tissue following adenoidectomy. DNA extracted from these samples was used for the amplification of 23S ribosomal RNA gene of HP by real-time polymerase chain reaction (RT-PCR). RESULTS: In the first group 34 effusion samples were obtained from ears of 18 patients (two had unilateral OME). HP was found to be positive in 12 children (67%) and 16 of 34 ears (47%) with RT-PCR. In eight children HP was positive in only one ear and in four children in both ears. No positive reaction was seen in tissue samples obtained from adenoids of these patients. In the second group a positive reaction was seen in adenoid tissue of only one patient. CONCLUSIONS: This study showed that there is HP presence in middle ears of the children with chronic OME, indicating HP having a possible role in OME pathogenesis. In addition, we demonstrated HP presence in only 1 of 38 adenoid specimens supporting the idea that adenoid tissue does not act as a reservoir for HP. PMID- 15885327 TI - Short use of muscle relaxants following single stage laryngotracheoplasty in children. AB - OBJECTIVE: The postoperative management of children undergoing single stage laryngotracheoplasty (SSLTP) includes intubation and muscle paralysis to secure the airway and protect the wound. We reduced the period of postoperative muscle paralysis in an attempt to decrease the incidence of pulmonary complications. The objective of this study was to evaluate the influence of the duration of muscle paralysis on pulmonary complications and outcome. METHODS: Medical records of all children admitted, between 1994 and 2002, to the pediatric intensive care unit following SSLTP were analysed. Children were grouped according to the number of days muscle paralysis was used. RESULTS: Thirty-six children (15 male, 21 female, mean age 32 months (9-162 months)) underwent SSLTP for laryngeal stenosis. Prior to surgery 29 needed a tracheotomy (mean duration 11.1 months). Shorter muscle paralysis leads to shorter intubation and mechanical ventilation and therefore PICU and hospital length of stay were 12.4 and 9.9days shorter in the group with short use of muscle paralysis (p<0.001 and p=0.002, respectively). There was no significant difference in postoperative complications, but a trend towards fewer atelectases in children with short muscle paralysis could be recognised. Postoperatively we observed no auto-extubations in either group and success rate of SSLTP was comparable in both groups (94 and 95%). CONCLUSION: Children undergoing SSLTP can safely benefit from a postoperative strategy using a short duration of muscle relaxants. They have fewer days on mechanical ventilation with a concomitant decrease in duration of hospital stay. PMID- 15885328 TI - Effects of otitis media with effusion on auditory temporal resolution. AB - OBJECTIVE: There is mounting evidence to suggest that otitis media with effusion (OME) is associated with auditory processing deficits that persist beyond the resolution of the peripheral hearing loss. This study investigated the residual effects of OME on auditory temporal resolution. EXPERIMENT 1: METHODS: Experiment 1 measured detection thresholds for a brief tone presented either before (backward masking) or during (simultaneous masking) a masking noise, in 6- and 8 year-old children. Six-year-olds were selected from a prospectively studied group with a lifetime known history of OME. Eight-year-old children, with a retrospectively determined history of OME, were also recruited. All children were free of OME at the time of testing. RESULTS: Regardless of OME history, 6-year old children had similar tone thresholds on all masking tasks. In contrast, 8 year-olds with a history of recurrent OME had 18 and 4dB higher mean thresholds for the backward and simultaneous masking conditions, respectively, compared with age-matched controls. Possible explanations for these results included (i) recruitment bias, rather than OME, contributed to differences in auditory processing abilities amongst 8-year-old children, or (ii) OME impaired performance at both ages, but this was not seen in 6-year-olds due to 'ceiling' effects. EXPERIMENT 2: METHODS: To distinguish between these possibilities, Experiment 2 measured temporal resolution, using backward masking and amplitude modulation detection, in the prospectively studied group of children when they were 8 years old. RESULTS: Regardless of OME history, these 8-year-olds had similar auditory temporal processing abilities. Results from Experiment 2 suggested that recruitment bias was the most likely explanation for the difference in auditory processing abilities between 8-year-old children with and without a history of OME found in Experiment 1. Consistent with previous data, associations were found between backward masking, age and cognitive ability. CONCLUSION: There is no evidence to suggest that OME effects temporal resolution after the recovery of normal pure-tone thresholds. PMID- 15885329 TI - Speech recognition abilities in noise for children with severe-to-profound unilateral hearing impairment. AB - OBJECTIVE: The purpose of this research was to compare the signal-to-noise ratio required for equal performance for children (aged 6-14 years) with normal hearing (N=17) versus those with severe-to-profound unilateral hearing impairment (N=20) who can be at a disadvantage, particularly when sounds are presented to the impaired ear. The listening environment is designed to approximate that which is encountered in a typical classroom. METHODS: All signals (Hearing in Noise Test Children and Nonsense Syllable Test) were presented in the sound field from various azimuths with continuous noise presented from all quadrants. The children were required to repeat twenty items, from each test, in each listening condition. The intensity of the presentations was varied based on the accuracy of previous items. Average signal-to-noise ratios are compared between and within groups. RESULTS: (1) In most listening conditions, significantly greater signal to-noise ratios were needed by those with unilateral hearing impairment than those with normal hearing, on both speech tests. (2) In every listening condition, both groups required significantly greater signal-to-noise ratios to perform equally well on the Nonsense Syllable Test as on the Hearing-In-Noise Test-Children. (3) For the Hearing-In-Noise Test-Children, children with normal hearing required significantly greater signal-to-noise ratios when facing the signal than when the signal was presented to a normally hearing ear (monaural direct). (4) On both tests, the children with unilateral hearing impairment required significantly greater signal-to-noise ratios when facing the signal than in the monaural direct condition. (5) On both tests, the children with unilateral hearing impairment required significantly greater signal-to-noise ratios when the signal was presented to the impaired ear (monaural indirect) than when facing the signal. CONCLUSIONS: (1) The children with unilateral hearing impairment require a more advantageous listening condition to perform equally as well as their normally hearing counterparts. (2) All of the children benefited from signals delivered in a monaural direct condition. (3) The children with unilateral hearing impairment performed best in the monaural direct condition or when facing the signal at zero degrees. (4) Significantly greater signal-to-noise ratios were needed for both groups when restricted contextual cues were available versus when sentential cues were available. PMID- 15885330 TI - Temporal bone histopathologic findings in partial trisomy 13 and partial trisomy 14. AB - OBJECTIVE: To describe temporal bone histopathology in an infant with partial trisomies of chromosomes 13 and 14. METHODS: Temporal bones were taken at autopsy from a 7-day-old neonate who has both partial trisomy 13 and partial trisomy 14. The right temporal bone was embedded in celloidin and sections were cut for microscopic examination. The left temporal bone was studied by microdissection. The middle ear was examined and the inner ear sensory organs dissected for study by light microscopy. RESULTS: The external auditory canal was stenotic in both ears. Remnants of mesenchymal tissue were present in the middle ear cavity. The middle ear ossicles were normal except that both stapes were malformed with a single crus and a small footplate. Both facial nerve canals were dehiscent in the region of the oval window. The cochlea was malformed bilaterally; a scala communis was present and the basilar membrane was abnormally short. No loss of sensory cells was observed in either cochlea. Blood vessels were found traversing scala vestibuli and there were cystic lesions in the stria vascularis and spiral ligament. In the middle cochlear turn, the bony wall of scala vestibuli and the osseous spiral lamina were covered by a substantial layer of connective tissue which appeared to be an extension of the spiral ligament. This is an unusual finding which, to the authors' knowledge, has not been previously reported. In the vestibular apparatus a wide communication was present between the saccule and utricle. CONCLUSIONS: As this study demonstrates, abnormalities of the external, middle, and inner ear may occur in cases of partial trisomy 13 and partial trisomy 14. Both temporal bone findings and clinical features in partial trisomy 13 and partial trisomy 14 to some degree overlap with those of trisomy 13, partial trisomy 13 and partial trisomy 14. PMID- 15885331 TI - Perceived qualitative benefits of cochlear implants in children with multi handicaps. AB - OBJECTIVE: To determine families' perceived benefits of cochlear implants in children with multiple-handicaps. METHODS: Parents/guardians of a child with a cochlear implant for at least 3 months, under the age of 18 years and had least one additional disability were recruited for participation through rehabilitation therapists. Families who did not have a child enrolled in therapy at the study center were contacted by mail. Families were asked a number of open-ended and close-ended questions. Interviews were audiotaped and transcribed, with answers to open-ended questions coded by themes. RESULTS: We interviewed 19 families of 20 children and included 15 families of 16 children who had had at least 6 months of cochlear implant experience for this study. The mean time between implantation and study was 3 years (S.D.+/-2.2) with a range of 0.5-8 years. Children were highly compliant in wearing their device. Sixty-eight percent (11/16) of the children were enrolled in additional therapies beyond speech and aural rehabilitation therapies. Fifty percent of families (8/16) indicated they had no obstacles to accessing therapies. Insurance was the most common barrier to accessing therapies (18% of respondents). All children made communication progress post-implant as described by their families. All families felt that the cochlear implant team provided enough information prior to receiving the implant to help families make the appropriate decision about implanting their child. All families said that if they were to make the decision again, they would choose to have their child implanted. CONCLUSION: More children with multiple-handicaps are receiving cochlear implants. Very young children may undergo cochlear implantation only to be subsequently diagnosed with an additional impairment or disability at later ages. As a result, it is important to understand the specific needs of this population and the parental perceptions of benefit in order to provide the best services and opportunities for success with a cochlear implant. Although not all of the children gained gold standard speech and language outcomes, all children broadened their communication skills. In motivated families of children with additional disabilities, it is appropriate to provide them with the same opportunity to access audition and expand their communication abilities as any other child with a hearing impairment. PMID- 15885332 TI - Microbiology of middle ear effusions from 292 patients undergoing tympanostomy tube placement for middle ear disease. AB - OBJECTIVE: Otitis media is one of the most commonly diagnosed childhood illnesses. Ideally, culture directed therapy for otitis media would be available, however, the common approach is to treat infections with antibiotics that cover the most common pathogens. The objective of this study is to describe the pathogens cultured from the middle ear effusions (MEE) of patients that underwent tympanostomy tube placement for middle ear disease, compare these results with previous studies, and assess for trends suggestive of changes in the microbiology of these patients. METHODS: Patients were invited to participate after the decision to place ventilation tubes had been made. A standard anterior-inferior myringotomy was made for placement of the ventilation tube. After myringotomy, patients had their ears suctioned and all effusions were collected for microbiologic analysis. RESULTS: A total of 292 patients were enrolled in the study, a total of 270 MEE samples were taken. Haemophilus influenzae and Moraxella catarrhalis were seen in 24 of 148 (16.2%) and 15 of 148 (10.1%), respectively. Neisseria meningitidis, Staphylococcus aureus and Corynebacterium species were each identified in 11 of the 148 pathogens isolated (7.4%). Streptococcus pneumoniae was identified in 6 of the 148 isolated pathogens (4.1%). CONCLUSION: Despite a high number of non-pathogenic bacteria, and a high number of culture negative effusions, we demonstrated pathogen rates consistent with other reports in the literature. Future directions include using more specific techniques such as PCR to better understand the microbiology of middle ear effusions. PMID- 15885333 TI - Management of antenatally detected fetal airway obstruction. AB - Five cases of antenatally diagnosed fetal airway obstruction have been cared for at the John Hunter Children's Hospital, Newcastle, Australia. A multidisciplinary team manages them during the perinatal period. We present our technique at the time of delivery, which aims to afford us the greatest flexibility in managing both the mother, her child's airway, and the underlying lesion. We begin with an ex utero intrapartum technique (EXIT) and favor routine rigid bronchoscopy to secure the neonate's airway without preliminary attempts at endotracheal intubation. PMID- 15885334 TI - Does topical lidocaine with adrenaline have an effect on morbidity in pediatric tonsillectomy? AB - OBJECTIVE: To evaluate the efficacy of lidocaine with adrenaline on post operative morbidity in pediatric patients after tonsillectomy. STUDY DESIGN: A double blind prospective randomized controlled clinical study. METHODS: The study is consisting of two groups of pediatric patients following tonsillectomy performed in a university hospital. One group received lidocaine with adrenaline soaked swabs packed in their tonsillar fossae while the control group received saline-soaked swabs. Chi-square and two-tailed unpaired Student's t-tests were used to compare the two independent groups. p<0.05 was accepted as statistically significant. RESULTS: No significant pain-relieving effect was seen in the lidocaine with adrenaline group (p>0.05) and also the other post-operative parameters such as nausea, fever, vomiting, odor, bleeding, otalgia and trismus were not statistically different between the two groups based on chi-square analysis (p>0.05). There were no complications associated with lidocaine and adrenaline. CONCLUSION: We suggest that application of topical lidocaine with adrenaline seems to be a safe and easy medication for local anesthetic use. However, in our study, lidocaine with adrenaline offered no advantage over placebo in the control of post-operative pain and other morbidity related factors following pediatric tonsillectomy. We therefore do not recommend topical application of lidocaine with adrenaline for reducing morbidity in pediatric tonsil surgery. PMID- 15885335 TI - Idiopathic sudden sensorineural hearing loss in children. AB - OBJECTIVE: Although idiopathic sudden sensorineural hearing loss (ISSHL) is a frequent disease in adults, less is known about incidence and treatment of ISSHL in children. METHOD: A retrospective chart analysis was performed to evaluate the frequency of ISSHL in children aged under 18 years between 2000 and 2003, who were treated in our department. Children received prednisolone intravenously at an initial dose of 3mg/kg bodyweight. Prednisolone dose was reduced to half every second day. The medication was given for a maximum of 14 days or finished 2 days after the hearing normalized in pure-tone audiometry. The follow-up was continued between 3 and 14 months. RESULTS: The complete recovery rate was 57%, and the partial recovery was 36%. Initial hearing loss of 50dB and more was predictive for poor outcome in children (p=0.028). Presence of tinnitus was without relevance for the outcome. The incidence of ISSHL in the local area of about 250,000 inhabitants was 1/10,000 in children. CONCLUSION: ISSHL seems to be a less frequent disease in children than in adults. Severe initial hearing loss is coupled with poor outcome. Under treatment with prednisolone hearing improvement was found in 13 of 14 patients. PMID- 15885336 TI - Oxidant and antioxidant levels in children with acute otitis media and tonsillitis: a comparative study. AB - OBJECTIVE: Recurrent episodes of acute otitis media (AOM) and acute tonsillitis (AT) are a common problem in infectious disorders during childhood and are major cause of morbidity in children. The organism maintains defense systems including nonenzymatic antioxidants such as Vitamins A, E and C and reduced glutathione (GSH) against reactive oxygen species (ROS). In the present study, lipid peroxidation status and nonenzymatic antioxidant capacity were investigated in children with AOM and AT. Our aim was to compare the lipid peroxidation and responses of the body's antioxidant status in the closely associated infections such as AOM and acute tonsillitis. METHODS: The study included 23 (14 males, 9 females) children with AOM, 27 (14 males, 13 females) with AT and 29 (16 males, 13 females) healthy control subjects. The ages of the study and control subjects were between 2 and 7 years. Serum beta-carotene, retinol, Vitamin E, Vitamin C, and whole blood malondialdehyde (MDA) (as an indicator of lipid peroxidation) and GSH levels were studied in all subjects. RESULTS: There was a statistically significant difference between the groups for all parameters (P<0.05). All of the antioxidant vitamins such as beta-carotene, retinol, Vitamin E, and Vitamin C levels were observed to be significantly decreased in the both patient groups. Nevertheless, GSH levels were also decreased in the patient groups. MDA levels were found to be higher in children with AOM and AT than in the healthy control subjects. When compared the AOM and AT groups, there was statistically significant difference between the groups for whole blood MDA (P<0.05). CONCLUSION: Even though they seem to react in a similar way against ROS, in the cases of AOM and AT, it is possible to see the different approaches in these tissues for the oxidative stress. PMID- 15885337 TI - Study of the hearing loss in children and adolescents, comparing the periods of 1990-1994 and 1994-2000. AB - INTRODUCTION: In 1994, a study was performed with 200 children and adolescents suffering from hearing loss. It concluded that the diagnostic confirmation of hearing loss within 2 years of age occurred in just 13% of the cases, although 56% were suspected in that phase. The loss of time of over 2 years between suspicion and confirmation of hearing loss occurred in 42% of the cases. OBJECTIVES: The comparison of the main hearing loss etiologies-genetic cause, consanguinity, congenital rubella, meningitis, perinatal events and unknown causes-in children and adolescents in the periods of 1990-1994 and 1994-2000; comparison of incidence, in males and females, for each etiology and among the others; comparison of age at the first consultation, for each and among them; and the investigation as to whether the time between suspicion and diagnosis of hearing loss was different for each etiology and among the others. METHODS: During the period of 1990-2000, of the 519 children and adolescents with hearing loss, 442 individuals were selected, in the two moments of the study: 1990-1994 and 1994-2000. The variables used were: sex, age at first consultation, suspected etiology and time between suspicion and confirmation of hearing loss. RESULTS: Congenital rubella, genetic and perinatal causes, meningitis, consanguinity and unknown causes were responsible for over 80% of all etiologies, in both periods. There were no differences between the sexes in the periods studied. There was no relation among age, sex and etiology. Among the etiologies studied, there were no differences in the lengths of times between suspicion and confirmation of hearing loss, in each period separately. The comparative study showed that congenital rubella, genetic and unknown causes took longer times between suspicion and confirmation of hearing loss, for the period of 1990-1994, as compared with 1994 2000. CONCLUSIONS: Congenital rubella remains as an important etiology, as well post-meningitis deafness. Age at first consultation did not show relationship to the hearing loss etiology nor to sex. Independently of whether the etiology being pre-natal, perinatal or post-natal, congenital or acquired, the length of time between suspicion and confirmation of hearing loss did not differ between the periods studied, separately. PMID- 15885338 TI - Neonatal hearing screening with transient evoked otoacoustic emissions in Western Saudi Arabia. AB - OBJECTIVE: To study the incidence of congenital hearing impairment in the Saudi population and to evaluate the need of establishing a Saudi universal neonatal hearing screening program based on transient evoked otoacoustic emission. METHODS: A total of 11986 well non-high-risk neonates were screened by transient evoked otoacoustic emission over period of 8 years from September 1996 to February 2004. The universal hearing screening was consequently done in a daily base before discharge from nursery. Those who failed the initial screening were followed up diagnostically until hearing loss was confirmed or excluded. RESULTS: From the total number of 11,986 neonates (41.4% male and 58.6% females) examined in this study 10943 (91.3%) neonates passed the first screening step while 1043(8.7%) neonates failed. From the 1043 neonates examined in the second screening step in the 5th day of life, 300 (29%) neonates failed. At the age of 5 months, all the 300 infants that failed the second screening step underwent a comprehensive audiological assessment to confirm the existence of hearing loss. The 278 infants that passed the assessment were considered as normal; while 22 failed and were confirmed to have congenital hearing loss. Of these 22 infants, 2 had unilateral sensorineural hearing loss, and the remaining 20 had bilateral sensorineural hearing loss. The incidence of sensorineural hearing loss was estimated to be 0.18% while the incidence of bilateral sensorineural hearing loss was 0.17%. No significant difference between males and females was found. The average age at confirmation of congenital hearing loss was 5.5 months. CONCLUSION: The incidence of congenital hearing loss in the western region of Saudi Arabia is relatively high compared with international figures. Hearing screening for all neonates using transient evoked otoacoustic emission should be part of the standard medical care in Saudi Arabia. PMID- 15885339 TI - The role of the anterior cricoid split in facilitating extubation in infants. AB - OBJECTIVE: The use of the anterior cricoid split (ACS) procedure has facilitated the extubation of children with early subglottic stenosis who may otherwise have required a tracheostomy. Criteria have been established for the evaluation of patients suitable for ACS and adherence to these guidelines has increased the extubation rate and reduced mortality. The objective of the study was to assess the use of the procedure at our tertiary referral centre. METHOD: In this retrospective clinical study, 33 patients were identified over an 11-year period between 1993 and 2004 with subglottic narrowing at the level of the cricoid ring, and who subsequently underwent an ACS. Demographic data, duration of intubation, indication and number of reintubations, success rate and complications were noted. Patients included in the study were infants who repeatedly failed trials of extubation and those who presented with persistent stridor. RESULTS: Twenty nine out of 33 (88%) children were successfully extubated as a result of the cricoid split procedure. Four children failed extubation after the cricoid split. Three required a tracheostomy and one child remained intubated for a prolonged period. CONCLUSION: The anterior cricoid split procedure has revolutionised the management of early subglottic stenosis in selected neonates with failed extubation. It is a safe operation, and if successful it avoids the formation of a tracheostomy. PMID- 15885340 TI - Calcified fibromatosis of the neck in 4-year old girl: rapid growth, rapid therapy. AB - Fibromatosis is a rare soft tissue disease typical for infants and characterized by fibroblastic proliferation, which may appear similar to fibrosarcoma. An unusual case of 4-year old girl presenting large tumor of the neck with massive calcification is described. The growth of the tumor was rapid and mediastinal involvement was observed. The final diagnosis showed benign nature of tumor with microscopic features of fibromatosis calcificans. After surgical resection of neck tumor, residual mass persisted both in the neck and in the mediastinum in 6 years of follow-up without signs of progression. PMID- 15885341 TI - Accidental cyanoacrylate glue ingestion. AB - Cyanoacrylates have a variety of medical and commercial applications as adhesives. They are commonly found as glue in the household. They can be swallowed accidentally by children. However, no case was reported so far in the English medical literature. Their effect on the mouth, pharynx, larynx, esophagus and the rest of gastrointestinal system is unknown. Here we report our own child who accidentally ingested cyanoacrylate glue without unwanted sequelae due to prompt intervention. PMID- 15885342 TI - Topical antibiotic induced otomycosis. AB - Prior to 1999, the diagnosis of otomycosis as a cause of persistent otorrhea was rare. An increase incidence has been seen in among our outpatient pediatric otolaryngology practice. The purpose of this study is to assess the contribution of ototopical antibiotic drops to the development of otomycosis. DESIGN: Retrospective study. SETTING: Pediatric otolaryngology outpatient center. METHODS: Chart review of all patients diagnosed with otomycosis between June 1999 and September 2001. Twenty-six patients (ages 17 months-29 years) were diagnosed with otomycosis based on clinical and microbiological findings after treatment with topical ofloxacin antibiotic drops. All patients had used ototopical antibiotics, including ofloxacin in every case, for presumed bacterial otorrhea. Once the fungal source was recognized, therapy succeeded in each case (26/26). Physicians need an elevated suspicion of otomycosis as a cause of persistent otorrhea, especially following treatment with topical antibiotic drops. Appropriate treatment of otomycosis eliminates otorrhea. Ofloxacin remains an excellent choice for bacterial otorrhea, but it appears to increase the incidence of otomycosis. Thus, its usage warrants careful post-treatment follow-up. PMID- 15885343 TI - Recurrent inverted papilloma of a pediatric patient: clinico-radiological considerations. AB - Inverted papilloma (IP) of the nasal cavity and paranasal sinuses is an uncommon pathology, especially in the pediatric population. We present a pediatric case of recurrent IP in the left nasal cavity. The physical examination of a 9-year-old girl revealed a polypoid mass filling the middle meatus in the left nasal cavity. Purulent secretion and adenoid vegetation obstructing the choanea were also noted. Coronal and axial maxillo-facial computed tomography demonstrated a mass in the nasal cavity, enlarging the left middle meatus. We discussed the imaging findings and treatment of IP and concluded that it should have been included in the differential diagnosis of nasal mass lesions in the pediatric population. PMID- 15885344 TI - Infantile myofibromatosis: unusual diagnosis in an older child. AB - This manuscript describes the unusual presentation of infantile myofibromatosis (IM) in an older child with its diagnosis and management. An 8-year-old girl presented with a painless, rapidly expanding malar mass. CT demonstrated an erosive soft tissue lesion and needle biopsy was nondiagnostic. Complete excision returned the pathologic diagnosis of IM. The patient had no complications and no evidence of recurrence at 1 year. Almost 90% of IM cases present by age two and IM in older children is highly unusual. The solitary form of IM is most common and its treatment is complete excision with an excellent prognosis. PMID- 15885345 TI - Successful laser ablation of diffuse laryngeal swelling in posttransplant lymphoproliferative disorder: the Japanese first case. AB - Posttransplant lymphoproliferative disorder (PTLD) is a well-known complication of pediatric organ transplantation. The first symptoms are often in the ear, nose, or throat (ENT) area. We report the first Japanese case of an airway obstruction in PTLD. A 7-year-old boy who had a liver transplant at the age of 2 years presented with progressive stridor. Fiberoptic examination revealed enlarged epiglottis and swelling arytenoids. Highly suspicious of early-PTLD, he was treated with reduction of immunosuppression, but no improvement on his larynx was observed. Laser ablation of swelling arytenoids worked. Surgical debulking is effective to ease an airway obstruction in cases of PTLD. PMID- 15885347 TI - Chronic hepatitis C, depression and interferon. PMID- 15885348 TI - Developing concepts on MELD: delta and cutoffs. PMID- 15885349 TI - Prevention of interferon-alpha associated depression in psychiatric risk patients with chronic hepatitis C. AB - BACKGROUND/AIMS: Interferon-alpha (IFN-alpha) induced depression is a major limitation for the treatment of chronic hepatitis C, especially for patients with psychiatric disorders. We prospectively studied the efficacy of a pre-emptive treatment with the antidepressant citalopram to prevent depression during hepatitis C treatment with pegylated IFN-alpha-2b plus ribavirin. METHODS: 14 HCV infected patients with psychiatric disorders received a prophylactic medication with citalopram (20mg/day) before and during therapy with IFN-alpha. The incidence of major depression was compared with 22 HCV infected patients with psychiatric disorders (group B; n=11) and without psychiatric risk factors (group C; n=11), who underwent IFN-alpha treatment without a pre-emptive antidepressant therapy. Depression was diagnosed by DSM-IV criteria. RESULTS: Pre-treatment of psychiatric patients with citalopram significantly reduced the incidence of major depression during the first 6 months of antiviral treatment as compared to the two control groups (group A 14% vs. 64% and 55% in group B and C; log-rank 6.89; df=2; P=0.032). Patients who developed symptoms of major depression during IFN therapy could also be improved by antidepressive treatment. CONCLUSIONS: Our open label pilot study, though small, clearly indicates that IFN alpha induced depression in psychiatric risk patients can be ameliorated by both the use of antidepressants as well as by intensive psychiatric care. However, larger, double blind placebo controlled trials in other patient populations are required to confirm these preliminary findings. PMID- 15885350 TI - Hepatitis B virus infection in lymphatic tissues in inactive hepatitis B carriers. AB - BACKGROUND/AIMS: Hepatitis B virus (HBV) infection in extrahepatic tissues is controversial. To clarify whether episomal HBV can infect nonhepatic tissues, we investigated the molecular forms of HBV in the lymphatic cells of inactive HBV carriers who lacked viremia, thus avoiding contamination with HBV genomes originating from the viral particles present in the serum. METHODS: We assessed HBV genome, replicative forms, and viral integrants in the liver, serum, peripheral blood mononuclear cells (PBMC), and lymph nodes of 21 inactive HBV carriers who tested positive for antibodies against the HBV core antigen (anti HBc). RESULTS: Of the 21 anti-HBc positive individuals, HBV-DNA was detected in liver samples of 15 (71.4%), in the lymph nodes of 11 (52.4%), and in PBMC of three (14.3%). However, none of the detected HBV genomes from lymphatic tissues included the replicative forms of HBV. In one case, integrated HBV was present in the lymphatic tissues and the host-viral junction was present in the intronic sequences of chromosome 17. CONCLUSIONS: These data suggest that human lymphatic tissues cannot support viral replication in anti-HBc positive inactive HBV carriers, while retaining the viral genome as an integrated form. PMID- 15885351 TI - Occult hepatitis B virus infection in hematopoietic stem cell donors in a hepatitis B virus endemic area. AB - BACKGROUND/AIMS: The acquisition of hepatitis B virus (HBV) infection following organ transplantation from donors with occult HBV infection is an important cause of morbidity and mortality. The aim of this study is to determine the prevalence of occult HBV in allogeneic hematopoietic stem cell (HSC) transplantation donors. METHODS: We performed a retrospective study on 124 consecutive hepatitis B surface antigen negative HSC donors. Their serum samples were analyzed by PCR for the pre-S/S, pre-core/core and X regions of the virus. Samples reactive by at least two PCR assays were considered HBV-DNA positive. RESULTS: Nineteen of the 124 HSC donors (15.3%) had occult HBV infection. Sixteen of these 19 donors with occult HBV infection (84.2%) tested positive for hepatitis B core antibody while 78 of 105 subjects (74.3%) without occult HBV infection were also positive (P=0.56). Fourteen of the 19 donors (73.7%) with occult HBV infection tested positive for hepatitis B surface antibody while 67 of the 105 subjects without occult HBV infection were also positive (P=0.45). CONCLUSIONS: The prevalence of occult HBV infection among HSC donors in Hong Kong is high. Anti-HBc and anti-HBs status had no significant correlation with the presence of occult HBV infection. PMID- 15885352 TI - MELD score and hepatocellular carcinoma identify patients at different risk of short-term mortality among cirrhotics bleeding from esophageal varices. AB - BACKGROUND/AIMS: The role of model for end stage liver disease (MELD) and the presence of hepatocellular carcinoma (HCC) as risk factors of short-term mortality in patients bleeding from oesophageal varices were evaluated. METHODS: From February 2002 to August 2003, 172 cirrhotic patients admitted for the first episode of bleeding from oesophageal varices received vasoactive and endoscopic therapy. Patients' survival was evaluated at 6 weeks and 3 months. The role of MELD and HCC as independent risk factors of mortality was evaluated. RESULTS: In the 172 patients, the overall mortality was 21.5% at 6 weeks and 30.2% at 3 months. MELD score resulted a good predictor of mortality either at 6 weeks or 3 months. Fifty-four patients (31.3%) had HCC. The presence of advanced HCC was an independent risk factor of mortality at 3 months. Patients with MELD score>15 and advanced HCC had a significantly worse survival than patients with MELD2.5 was the only significant prognostic predictor at 6 (odds ratio: 9.8, P<0.001) and 12 months (odds ratio: 16.3, P<0.001) in multivariate logistic analysis. CONCLUSIONS: Increasing MELD score is associated with the onset of ascites and encephalopathy. DeltaMELD is superior to initial MELD and CTP scores to predict intermediate term outcome in patients with advanced cirrhosis. PMID- 15885354 TI - Cloning and characterization of hepaCAM, a novel Ig-like cell adhesion molecule suppressed in human hepatocellular carcinoma. AB - BACKGROUND/AIMS: Previously, we reported on gene HEPN1 that was silenced in hepatocellular carcinoma (HCC) and its capability of arresting cell growth. In this study, we identified another novel gene hepaCAM from the liver, which contains the full-length HEPN1 on its antisense strand in the 3'-noncoding region, and assessed its expression, characteristics and functions in HCC. METHODS: Full-length hepaCAM cDNA was isolated by rapid amplification of cDNA ends. The gene expression was examined by semi-quantitative RT-PCR in 23 paired HCC liver specimens and 5 HCC cell lines. Transfection studies, coupled with immunocytochemistry, cellular interaction analyses, colony formation and microtetrazolium assay, were employed to elucidate the localization and functions of hepaCAM. RESULTS: The expression of hepaCAM decreased in 20/23 of HCC samples and was undetectable in 5 HCC cell lines tested. The gene product consisting of 416 amino acids displayed the typical structure of Ig-like cell adhesion molecules. The protein was glycosylated and predominantly localized on the cytoplasmic membrane. When re-expressed in HepG2, hepaCAM accelerated cell spreading (P<0.001), increased cell motility (P=0.0011), reduced colony formation (P=0.0022), and inhibited cell growth (P<0.001). CONCLUSIONS: Gene hepaCAM, frequently silenced in HCC, encodes an Ig-like transmembrane glycoprotein and is involved in cell adhesion and growth control. PMID- 15885355 TI - Activation of the canonical Wnt/beta-catenin pathway confers growth advantages in c-Myc/E2F1 transgenic mouse model of liver cancer. AB - BACKGROUND/AIMS: Previously, we showed that activation of the beta-catenin/Wnt pathway is a dominant event during c-Myc/E2F1 hepatocarcinogenesis. Majority of c Myc/E2F1 HCCs displayed nuclear accumulation of beta-catenin in the absence of beta-catenin mutations, suggesting that alterations in other members of the Wnt pathway might be responsible for nuclear localization of beta-catenin. Here, we investigated the mechanisms responsible for nuclear translocation of wild-type beta-catenin and addressed the potential contribution of the Wnt pathway in c Myc/E2F1 hepatocarcinogenesis. METHODS: Status of the members of the Wnt pathway was determined through microsatellite and Western blot analysis. RESULTS: Majority of c-Myc/E2F1 HCCs exhibited multiple abnormalities in the Wnt pathway regardless of the presence of beta-catenin mutations. The observed abnormalities included overexpression of Wnt-1, Frizzled 1 and 2 receptors, Dishevelled-1, downregulation of Secreted frizzled-related protein-1, GSK-3beta inactivation, microsatellite instability at the Axin locus as well as induction of beta-catenin target genes, such as glutamine synthetase, glutamate transporter-1, and Wisp-1. HCCs with beta-catenin activation displayed significantly higher proliferation rate and larger tumor size when compared with beta-catenin negative tumors. CONCLUSIONS: The data demonstrate that multiple abnormalities in the members of the Wnt pathway lead to nuclear accumulation of beta-catenin and suggest that activation of Wnt pathway provides proliferative advantages in c-Myc/E2F1-driven hepatocarcinogenesis. PMID- 15885356 TI - JNK mediates hepatic ischemia reperfusion injury. AB - BACKGROUND/AIMS: Hepatic ischemia followed by reperfusion (I/R) is a major clinical problem during transplantation, liver resection for tumor, and circulatory shock, producing apoptosis and necrosis. Although several intracellular signal molecules are induced following I/R including NF-kappaB and c-Jun N terminal kinase (JNK), their roles in I/R injury are largely unknown. The aim of this study is to assess the role of JNK during warm I/R injury using novel selective JNK inhibitors. METHODS: Male Wistar rats (200+/-25 g) are pretreated with vehicle or with one of three compounds (CC0209766, CC0223105, and CC-401), which are reversible, highly selective, ATP-competitive inhibitors of JNK. In the first study, rats are assessed for survival using a model of ischemia to 70% of the liver for 90 min followed by 30% hepatectomy of the non-ischemic lobes and then reperfusion. In the second study, rats are assessed for liver injury resulting from 60 or 90 min of ischemia followed by reperfusion with analysis over time of hepatic histology, serum ALT, hepatic caspase-3 activation, cytochrome c release, and lipid peroxidation. RESULTS: In the I/R survival model, vehicle-treated rats have a 7-day survival of 20-40%, while rats treated with the three different JNK inhibitors have survival rates of 60-100% (P<0.05). The decrease in mortality correlates with improved hepatic histology and serum ALT levels. Vehicle treated rats have pericentral necrosis, neutrophil infiltration, and some apoptosis in both hepatocytes and sinusoidal endothelial cells, while JNK inhibitors significantly decrease both types of cell death. JNK inhibitors decrease caspase-3 activation, cytochrome c release from mitochondria, and lipid peroxidation. JNK inhibition transiently blocks phosphorylation of c-Jun at an early time point after reperfusion, and AP-1 activation is also substantially blocked. JNK inhibition blocks the upregulation of the pro-apoptotic Bak protein and the degradation of Bid. CONCLUSIONS: Thus, JNK inhibitors decrease both necrosis and apoptosis, suggesting that JNK activity induces cell death by both pathways. PMID- 15885357 TI - Global gene repression in hepatocellular carcinoma and fetal liver, and suppression of dudulin-2 mRNA as a possible marker for the cirrhosis-to-tumor transition. AB - BACKGROUND/AIMS: Whether the transcriptional reprogramming process induced by hepatocellular carcinoma recapitulates that of the developing liver is at present unclear. METHODS: With a complete coverage of the liver transcriptome by microarray using adult livers as controls, we searched for similarities and differences in mRNA abundances between hepatocellular carcinoma nodules and fetal livers taken before (early) or after (late) the 22-24th week of gestation. Changes in some mRNA levels were studied in further liver samples by quantitative RT-PCR. RESULTS: Altered gene expression in hepatocellular carcinoma mostly results in down-regulated mRNAs which largely overlap with those repressed in the late fetal liver. Different frequencies of transcription factor binding sites in the down-regulated genes vs control genes as well as changes in abundance of mRNAs for relevant transcription factors point to a transcriptional repression. The down-regulated mRNAs code for proteins involved in (i) transcription and translation, (ii) specific functions of the differentiated hepatocyte or (iii) activation of proliferation and apoptosis. CONCLUSIONS: Apoptosis limitation is likely to predominate over active proliferation during liver development and hepatocellular carcinoma. Repression of the apoptosis-associated dudulin-2 mRNA points to a potential marker for the transition from a carcinoma-free to carcinoma-associated cirrhosis. PMID- 15885358 TI - Insulin prevents liver damage and preserves liver function in lipopolysaccharide induced endotoxemic rats. AB - BACKGROUND/AIMS: Liver integrity and function are crucial for survival of patients suffering from trauma, operations or infections. Insulin decreased mortality and prevented the incidence of multi organ failure and infection in critically ill patients. The aim of the present study was to determine whether insulin exerts positive effects on hepatic homeostasis and function during endotoxemia. METHODS: Endotoxemic rats received either saline or insulin. Hepatic morphology and function was determined by measuring the effect of insulin on liver proteins, enzymes, hepatocyte apoptosis and proliferation including caspases-3 and -9 and Bcl-2. Intrahepatic ATP, glucose and lactate concentration were determined by bioluminescence. To determine possible molecular changes the effect of insulin on hepatic cytokine mRNA and gene profile analysis were assessed. RESULTS: Insulin significantly improved hepatic protein synthesis by increasing albumin and decreasing c-reactive protein, P<0.05. Insulin attenuated hepatic damage by decreasing AST and ALT, P<0.05. Improved liver morphology was due to decreased hepatocyte apoptosis along with decreased caspase-3 concentration and increased hepatocyte proliferation along with Bcl-2 concentration, P<0.05. Insulin decreased hepatic IL-1beta, IL-6 and MIF mRNA and improved hepatic glucose metabolism and glycolysis, P<0.05. GeneChip analysis revealed an anti-inflammatory effect of insulin. CONCLUSIONS: Insulin improves hepatic integrity, hepatic glucose metabolism and hepatic function by increasing cell survival and attenuating the hepatic inflammatory response in endotoxemic rats. PMID- 15885359 TI - Interferon-alpha-induced modulation of glucocorticoid and serotonin receptors as a mechanism of depression. AB - BACKGROUND/AIMS: The mechanism of interferon (IFN)-alpha-induced depression remains poorly understood. Recently, modulation of glucocorticoid receptor (GR) and serotonin receptor 1A (5-HTR1A) were implicated in mechanism(s) leading to depression. To gain insight into this mechanism, we assessed the effect of IFN alpha on the modulation of GR and 5-HTR1A expression. METHODS: Hepatoblastoma, myelocyte-derived and T cell leukemia-derived cell lines were treated with titrated doses of IFN-alpha for different incubation times and analyzed by Western blot, RT-PCR, and microarrays. Dose- and time-dependent decreases of proteins and mRNA levels of GR and 5-HTR1A were observed. RESULTS: The expression of GR and 5-HTR1A in cells treated for 6 days decreased by 74 and 72%, respectively. Recovery was observed following IFN-alpha withdrawal. Co-incubation with tricyclic antidepressants (desipramine) or serotonin reuptake inhibitors (fluoxetine) attenuated the effect of IFN-alpha on GR or 5-HTR1A. GR and 5-HTR1A were unaffected by treatment with either IFN-gamma or tauroursodeoxycholic acid (TUDCA). However, the effect of IFN-alpha on GR was abolished when used in combination with TUDCA. CONCLUSIONS: In conclusion, IFN-alpha downregulated GR and 5-HTR1A levels in cell lines. These levels of GR and 5-HTR1A, following IFN alpha-induced downregulation, recovered after withdrawal of IFN-alpha or addition of desipramine or fluoxetine. These data provide insights regarding pathogenesis of IFN-alpha-induced depression. PMID- 15885360 TI - Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells--a potential mechanism for targeting liver anti-fibrotic therapeutics. AB - BACKGROUND/AIMS: Hepatic stellate cells are pivotal to fibrogenesis in the liver and many potential anti-fibrotic therapeutics are required to act on targets within hepatic stellate cells. The aim of this study was to generate a human antibody fragment to hepatic stellate cells. METHODS: Phage display was used to generate a human monoclonal antibody fragment to a peptide sequence present on an extracellular domain of synaptophysin, a protein expressed on the surface of hepatic stellate cells. RESULTS: An antibody fragment was isolated (termed C1-3), expressed in bacteria and purified. Fluorescently-labelled C1-3 antibody associated with human hepatic stellate cells but not hepatocytes in culture. Binding of fluorescently labelled C1-3 to hepatic stellate cells was blocked by the extracellular synaptophysin peptide sequence and uptake of the antibody intracellularly was inhibited by monensin. The toxin tributyl tin-when conjugated to C1-3-retained the ability to kill hepatic stellate cells confirming that C1-3 is sequestered intracellularly. CONCLUSIONS: This antibody fragment may be an effective means to target therapeutics to human hepatic stellate cells. PMID- 15885361 TI - A distinct microarray gene expression profile in primary rat hepatocytes incubated with ursodeoxycholic acid. AB - BACKGROUND/AIMS: Ursodeoxycholic acid (UDCA) and its taurine-conjugated derivative, TUDCA, modulate cell death and cell cycle regulators, such as E2F-1 and p53. However, precise pathways underlying UDCA's effects are not fully understood. The aim of this study was to identify specific cellular targets of UDCA. METHODS: The expression profile of primary rat hepatocytes incubated with UDCA was determined using Affymetrix GeneChip Rat 230A arrays. Hybridization data were processed to identify genes with significant expression changes. RT-PCR and immunoblot analyses of a selected target confirmed microarray data. RESULTS: The results showed that >440 genes were modulated with UDCA by >1.5-fold; approximately 25% were significantly different from controls. Genes affected by UDCA included new regulatory molecules, such as Apaf-1. RT-PCR and immunoblotting confirmed a decrease in Apaf-1. Other altered genes were directly involved in cell cycle (cyclin D1, cadherin 1, HMG-box containing protein 1) and apoptosis (prothymosin-alpha) events. The E2F-1/p53/Apaf-1 pathway appears to be targeted by UDCA. Finally, transcripts for proteins with kinase activity and transcription factors were specifically modulated by TUDCA. CONCLUSIONS: This study expands our knowledge of the biological effects of UDCA in hepatocytes. Most of the identified genes represent novel potential targets of UDCA, which may ultimately explain its therapeutic properties. PMID- 15885362 TI - Effects of betaine supplementation on hepatic metabolism of sulfur-containing amino acids in mice. AB - BACKGROUND/AIMS: We previously reported that acute betaine treatment induced significant changes in the hepatic glutathione and cysteine levels in mice and rats. The present study was aimed to determine the effects of dietary betaine on the metabolism of sulfur-containing amino acids. METHODS/RESULTS: Male mice were supplemented with betaine (1%) in drinking water for up to 3 weeks. Changes in hepatic levels of major sulfur amino acid metabolites and products were stabilized after 2 weeks of betaine supplementation. Betaine intake increased methionine, S-adenosylmethionine, and S-adenosylhomocysteine levels significantly, but homocysteine and cystathionine were reduced. Methionine adenosyltransferase activity was elevated to three-fold of control. Cysteine catabolism to taurine was inhibited as evidenced by a decrease in cysteine dioxygenase activity and taurine levels in liver and plasma. Despite the significant changes in the transsulfuration reactions, neither hepatic cysteine nor glutathione was altered. Betaine supplementation decreased the hepatotoxicity induced by chloroform (0.5 ml/kg, ip) significantly. CONCLUSIONS: Betaine supplementation enhances recycling of homocysteine for the generation of methionine and S-adenosylmethionine while reducing its utilization for the synthesis of cystathionine and cysteine. However, the hepatic levels of cysteine or glutathione are not affected, most probably due to the depression of taurine generation from cysteine. PMID- 15885363 TI - Association of myeloperoxidase promotor polymorphism with cirrhosis in patients with hereditary hemochromatosis. AB - BACKGROUND/AIMS: Hereditary hemochromatosis (HHC) is a disorder of iron metabolism with variable penetrance. Oxidative stress plays a central role in the progression to cirrhosis. Several enzymes involved in the production or degradation of reactive oxidants, like myeloperoxidase (MPO) and heme oxygenase (HO)-1 are influenced by promotor polymorphisms. This study assessed the impact of polymorphisms of the MPO (-463G/A) and the HO-1 promotors of Vienna (GT)n on the evolution of cirrhosis in patients with HHC. METHODS: One-hundred and fifty eight C282Y homozygotes without cofactors for fibrosis progression (119 males; mean age: 51.0+/-13.3) were studied. All patients underwent liver biopsy. Hepatic iron content was measured by atom absorption spectrophotometry. MPO polymorphism was assessed by RFLP analysis; HO-1 microsatellite polymorphism by a laser-based semi-automated DNA sequencer. RESULTS: The MPO genotypes GG, GA, and AA were found in 102 (64.6%), 45 and 11 patients, respectively. The GG-genotype was more common in patients with cirrhosis than in those without (78.7 vs. 55.7%, P=0.003). The distribution of HO-1 genotypes was not different. Logistic regression analysis revealed MPO genotype-GG, serum ferritin, age and male sex as independent predictors for cirrhosis. CONCLUSIONS: MPO genotype GG is associated with cirrhosis in patients with hereditary hemochromatosis. PMID- 15885364 TI - Immune-mediated liver injury. PMID- 15885365 TI - NASH: a mitochondrial disease. PMID- 15885367 TI - Are alcohol, tobacco and obesity genuine risk factors for hepatocellular carcinoma? PMID- 15885368 TI - The predictive value factor VII in acute hepatitis and coagulapathy. Is it really predictive? PMID- 15885370 TI - Cholestatic liver injury after glimepiride therapy. AB - Drug induced hepatotoxicity has been reported infrequently with sulfonylureas. For glimepiride, a second-generation sulfonylurea there is no report of hepatotoxicity in English literature. A patient with non-insulin-dependent diabetes mellitus who developed cholestatic liver injury soon after initiation of glimepiride therapy is presented. Complete work-up disclosed no other cause for hepatotoxicity including negative serological results for viral hepatitis. Liver biopsy was consistent with drug-induced cholestasis. The patient recovered 50 days after stopping glimepiride with no further recurrences. PMID- 15885371 TI - Aceruloplasminemia in an asymptomatic patient with a new mutation. Diagnosis and family genetic analysis. AB - A 39-year-old asymptomatic man showed elevated serum ferritin levels, mild hypertransaminasemia and serum ceruloplasmin almost undetectable. There was histological iron accumulation within the hepatocytes and also in the central nervous system (MRI). A genetic analysis revealed a new missense mutation in the ceruloplasmin gene. Two of the other four siblings were also affected by this mutation. PMID- 15885372 TI - Unusual appearances of oesophageal varices on imaging after sclerotherapy. PMID- 15885375 TI - Process-oriented dose assessment model for 14C due to releases during normal operation of a nuclear power plant. AB - Swedish nuclear utility companies are required to assess doses due to releases of radionuclides during normal operation. In 2001, calculation methods used earlier were updated due to new authority regulations. The isotope (14)C is of special interest in dose assessments due to the role of carbon in the metabolism of all life forms. Earlier, factors expressing the ratio between concentration of (14)C in air and in various plants were used. In order to extend the possibility to take local conditions into account, a process-oriented assessment model for uptake of carbon and doses from releases of (14)C to air was developed (POM(14)C). The model uses part of DAISY which has been developed to model the turnover of carbon in crops. [Hansen, S., Jensen, H.E., Nielsen, N.E., Svendsen, H., 1993. Description of the Soil Plant System Model DAISY, Basic Principles and Modelling Approach. Simulation Model for Transformation and Transport of Energy and Matter in the Soil Plant Atmosphere System. Jordbruksforlaget, The Royal Veterinary and Agricultural University, Copenhagen, Denmark]. The main objectives were to test model performance of the former method, and to investigate if taking site specific parameters into account to a greater degree would lead to major differences in the results. Several exposure pathways were considered: direct consumption of locally grown cereals, vegetables, and root vegetables, as well as consumption of milk and meat from cows having eaten fodder cereals and green fodder from the area around the nuclear plant. The total dose of the earlier model was compared with that of POM(14)C. The result of the former was shown to be slightly higher than the latter, but POM(14)C confirmed that the earlier results were of a reasonable magnitude. When full account of local conditions was taken, e.g. as regards solar radiation, temperature, and concentration of (14)C in air at various places in the surroundings of each nuclear plant, a difference in dose between sites of approximately one order of magnitude was found. PMID- 15885376 TI - Studies of Np and Pu in the marine environment of Swedish-Danish waters and the North Atlantic Ocean. AB - The long-lived anthropogenic radionuclides (237)Np, (239)Pu and (240)Pu were determined in marine environmental samples (seaweed and seawater) collected from Swedish-Danish waters and the North Atlantic Ocean at various locations on different occasions during the period 1991-2001. The measurements were performed with sector field Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and conventional alpha spectrometry. The (237)Np activity concentrations in Fucus vesiculosus and surface seawater from the Swedish west coast and Danish waters ranged from 0.16+/-0.02 to 1.02+/-0.09 mBq kg(-1) (dry weight) and 0.65+/-0.02 to 1.69+/-0.02 mBq m(-3), respectively, depending on the location and sampling year. Most of the (237)Np in these waters is believed to originate from the Sellafield nuclear reprocessing plant, with some contribution from global fallout. The (240)Pu/(239)Pu atomic ratios in F. vesiculosus samples are reported in this study with an overall average of 0.17+/-0.03. The (237)Np and (239)Pu activity concentrations observed in surface seawater collected in North Atlantic waters ranged from 0.16+/-0.01 to 0.62+/-0.08 mBq m(-3) and from 0.64+/-0.05 to 4.27+/ 0.08 mBq m(-3), respectively, and the (237)Np/(239)Pu atomic ratios were a good indicator of conservative behaviour of Np in marine waters. PMID- 15885377 TI - Investigation of 85Sr adsorption on selected soils of different horizons. AB - Studies on the mechanism of (90)Sr migration in soil require many processes to be considered. One of the most important is sorption on the surface of mineral components of the soil. In this study adsorption of (85)Sr on a variety of soil types from different horizons has been investigated. Adsorption isotherms show various affinities of (85)Sr, depending on soil type and to a lesser extent the horizon. An important effect of pH was found with a maximum in the range 5-7. The influence of calcium ions on the extent of adsorption of (85)Sr isotope on soil samples from surface horizons of four sites is presented. Depending on the soil type differing degrees of competitive adsorption of Sr and Ca were observed. Desorption of (85)Sr by distilled water as well as Ca(NO(3))(2) solution was also examined. Both methods resulted in the removal of a considerable proportion of the adsorbed isotope from the soil. Additionally the kinetics of the desorption process were studied. PMID- 15885378 TI - Radon exhalation of cementitious materials made with coal fly ash: Part 1- scientific background and testing of the cement and fly ash emanation. AB - Increased interest in measuring radionuclides and radon concentrations in fly ash, cement and other components of building products is due to the concern of health hazards of naturally occurring radioactive materials (NORM). The current work focuses on studying the influence of fly ash (FA) on radon-exhalation rate (radon flux) from cementitious materials. The tests were carried out on cement paste specimens with different FA contents. The first part of the paper presents the scientific background and describes the experiments, which we designed for testing the radon emanation of the raw materials used in the preparation of the cement-FA pastes. It is found that despite the higher (226)Ra content in FA (more than 3 times, compared with Portland cement) the radon emanation is significantly lower in FA (7.65% for cement vs. 0.52% only for FA). PMID- 15885379 TI - Radon exhalation of cementitious materials made with coal fly ash: Part 2- testing hardened cement-fly ash pastes. AB - Increased interest in measuring radionuclides and radon concentrations in fly ash (FA), cement and other components of building products is due to the concern about health hazards of naturally occurring radioactive materials (NORM). The paper focuses on studying the influence of FA on radon exhalation rate (radon flux) from cementitious materials. In the previous part of the paper the state of the art was presented, and the experiments for testing raw materials, Portland cement and coal fly ash, were described. Since the cement and FA have the most critical role in the radon release process relative to other concrete constituents (sand and gravel), and their contribution is dominant in the overall radium content of concrete, tests were carried out on cement paste specimens with different FA contents, 0-60% by weight of the binder (cement+FA). It is found that the dosage of FA in cement paste has a limited influence on radon exhalation rate, if the hardened material is relatively dense. The radon flux of cement-FA pastes is lower than that of pure cement paste: it is about approximately 3 mBq m(-2) s(-1) for cement-FA pastes with FA content as high as 960 kg m(-3). PMID- 15885380 TI - Geophysical methods in radon risk studies. AB - The results of the studies presented in the paper have shown that in the Upper Silesian Region in Poland, radon indoor concentration levels depend first of all on the geological structure of the subsurface layers. The essential factors influencing radon migration ability are the mining-induced transformations of a rock mass. In some cases, significant variations of radon potential have been found at sites featuring similar geological structures and experiencing comparable mining effects. To find out the causes of these variations, studies involving geophysical methods such as electrical resistivity profiling (PE) and electrical resisitivity sounding (VES) were used. These studies have shown that the measurements made using the electrical resistivity method can be helpful in evaluating radon potential of both the tectonically disturbed areas and the mining-transformed ones. PMID- 15885381 TI - Factors controlling measurements of radon mass exhalation rate. AB - Radon mass exhalation rate of soil samples was measured using an exhalation chamber of 10 dm(3) volume and a Lucas cell. The results show that mass of sample, grain size and water content influence the radon mass exhalation rate. For soil of (226)Ra activity concentration about 2500 Bq kg(-1) and samples within the range from 0.20 kg to 0.50 kg, the radon mass exhalation rate values are higher than those for samples of other masses. The observed radon exhalation rate is an inverse function of the average grain size. At the water content about 6% by weight, the radon mass exhalation rate reaches maximum, then it decreases with both increasing and decreasing of the water content in the sample. PMID- 15885382 TI - Distribution of uranium and radium isotopes in an aquifer of a semi-arid region (Manouba-Essijoumi, Northern Tunisia). AB - Groundwaters from the Sebkhet Essijoumi drainage basin, situated in northern Tunisia, West of the city of Tunis, were sampled and analyzed for uranium and radium isotopes. Low (234)U/(238)U activity ratios coupled with relatively high (228)Ra and (238)U concentrations were found in the Manouba plain phreatic aquifer, at the northern part of the basin, where remote sensing has indicated that this plain corresponds to the main humid zone of the area. Low (234)U/(238)U ratios probably reflected short residence time for waters in the Manouba plain, and high ratios longer residence time in the south, where water reaching the phreatic aquifer seems to have previously circulated in rocks constituting the southern hills. Assuming that, in the Manouba plain aquifer, the groundwater flows downstream from the Oued Lill pass area to the South-West of the Sebkha, the difference in the (228)Ra/(226)Ra activity ratio suggests that the residence time of water has been 2.8 years longer near the Sebkha than upstream. PMID- 15885383 TI - Intake of uranium and radium-226 due to food crops consumption in the phosphate region of Pernambuco--Brazil. AB - The phosphate region located in the Northeast of Brazil covers an area of approximately 150 km long with an average width of 4 km, along the coast of the states of Pernambuco and Paraiba. The inhabitants of this area are exposed to natural radioactivity levels higher than the background values recorded in the literature, mainly due to the presence of uranium and its decay products in the phosphatic sediments. The main aim of this study was to determine the activity concentration of uranium and (226)Ra in foodstuffs cultivated in this area, where the phosphate mineral has been extracted. The activity concentrations found for uranium and (226)Ra in the foodstuffs analyzed varied from 13 to 186 mBq kg(-1) (wet weight), with a mean value of 46 mBq kg(-1) and from 43 to 2209 mBq kg(-1) (wet weight), with a mean value of 358 mBq kg(-1), respectively. The annual intake of these radionuclides, for rural residents, was 7.45 Bq for uranium and 69.3 Bq for (226)Ra. PMID- 15885384 TI - Treatment of motor dysfunction in Parkinson's disease: an overview. AB - Levodopa remains the most effective treatment for Parkinson's disease (PD). However, the use of this drug is complicated by several adverse effects, in particular motor fluctuations and dyskinesias. Drugs providing more continuous dopaminergic stimulation and surgical approaches, such as deep-brain stimulation, have an important ameliorative effect on these problems. Despite these advances, the progression of the disease remains unaffected and strategies that slow or stop the neurodegenerative process are currently not available. Nevertheless, several compounds or surgical procedures are candidates for being neuroprotective and some of them are under evaluation. PMID- 15885385 TI - A new approach for the estimation of intervertebral disc volume using the Cavalieri principle and computed tomography images. AB - This study was carried out to describe a simple, accurate and practical technique for estimating the volume of intervertebral disc (ID) by the combination of the Cavalieri principle and computed tomography (CT) images. Total eight lumbar IDs from two cadavers were CT scanned in axial, sagittal and coronal sections. The consecutive sections with 5 and 3mm thickness were used to estimate the volume of the IDs by the Cavalieri principle. Three investigators estimated the volume of IDs independently to evaluate inter-observer differences. When the results were compared to the real volumes of IDs measured by the fluid displacement technique, there was no significant difference between the real volume measurements and the Cavalieri estimation results of ID volumes (P > 0.05). Moreover, findings of three investigators did not show significant variations (P > 0.05). Our results indicate that the section thickness and the section planes did not affect the accuracy of the disc volume estimation. Thus the combination of CT scanning with the Cavalieri principle may be used as a direct and reliable technique to estimate the volume of IDs with a mean of 4 min workload per ID. PMID- 15885386 TI - Functional outcome of intrathecal baclofen administration for severe spasticity. AB - PURPOSE: To estimate the functional benefit in patients with severe spasticity treated with intrathecal baclofen infusion through an implantable pump and to stress the need for functional assessment of these patients with a functional scale. PATIENTS AND METHODS: Between 1999 and 2003, 22 patients with a long history of severe and disabling pharmaceutically intractable spasticity, underwent implantation of a pump for continuous intrathecal baclofen infusion. The patients were subdivided into two categories according to the aetiology of spasticity: 15 had Multiple Sclerosis and seven had suffered a Spinal Cord Injury at different levels (from C4 to T11). Clinical status was assessed with the Ashworth and Penn spasm scales. Functional benefits were evaluated with the Barthel index score and pain relief with a self-reported visual analogue pain scale. RESULTS: Postoperatively, all patients presented improvement in spasticity, reduction of spasm frequency, significant improvement in functional status, enhancement of life comfort and reduction of pain. CONCLUSION: Reduction of spasticity and spasms achieved with intrathecally delivered baclofen, leads to functional improvement and pain relief. PMID- 15885387 TI - Tissue plasminogen activator expression in meningiomas and glioblastomas. AB - OBJECTIVES: Enzyme-linked immunosorbent assay (ELISA) and Western blotting techniques were used to investigate and compare the expression of tissue plasminogen activator (tPA) in benign (meningioma) and malignant (glioblastoma) human brain tumours. METHODS: A total of 22 tumour samples comprising 11 meningiomas and 11 glioblastomas with adjacent peritumoural tissue were analysed. RESULTS: The mean tPA content of meningiomas was approximately half that of glioblastomas (55.40 (S.D. 34.58) versus 106.98 (S.D. 43.82) ng/ml, p=0.006). Comparing tPA quantity in tumour and peritumoural tissue, there was a significant difference for meningiomas (55.40 (S.D. 34.58) versus 28.35 (S.D. 22.55) ng/ml, p=0.05), but no difference for glioblastomas (106.98 (S.D. 43.82) versus 84.23 (S.D. 57.39) ng/ml, p=0.32). Comparing tumour with normal brain tissue, there was no difference for meningiomas (55.40 (S.D. 34.58) versus 33.08 (S.D. 21.55) ng/ml, p=0.22), but a significant difference for glioblastomas (106.98 (S.D. 43.82) versus 33.08 (S.D. 21.55) ng/ml, p=0.004). Western blotting showed that in the meningioma group, the molecular weight pattern was constant with a dominant well-defined band at 41kD. Peritumoural tissue demonstrated two bands, with the stronger band at 41kD and a slightly weaker band at 71kD. In the glioblastoma group, there was more heterogeneity, with a dominant 41kD band found in all tumour and peritumoural samples, together with additional bands at 34, 58 and 66kD. CONCLUSION: These results indicate that (1) tPA is present in larger quantities in glioblastoma compared to meningioma and normal brain, (2) tPA quantity is not significantly different in the peritumoural tissue adjacent to glioblastoma but is significantly less for meningioma, and (3) tPA is expressed in more heterogenous forms in glioblastoma. This present study therefore suggests that the expression of tPA in a brain tumour may be an additional prognostic factor in terms of its malignant and invasive potential. PMID- 15885388 TI - Magnetic resonance angiography of the circle of Willis in migraine patients. AB - We analysed records of 244 consecutive adult outpatients who presented at our neurological polyclinic between 1996 and 2001 with migraine and who underwent a MRA of the circle of Willis. In 80 cases, a definitive migraine diagnosis according to the International Headache Society criteria (1988) could be retrieved from the patient records. Of these, 63 patients had migraine without aura and 17 patients had migraine with aura. Two MRA procedures were abrupted because of claustrophobia and another two MRA studies were unreliable because of movement artefacts. In only one patient of the remaining 240, an aneurysm was suspected with MRA. However, a conventional angiography did not confirm any cerebrovascular abnormality. All other exams were considered normal. This retrospective study argues against a systematic screening of migraine patients with MRA. PMID- 15885389 TI - Clinical features of 35 patients with Parkinson's disease displaying REM behavior disorder. AB - OBJECTIVE: To assess and compare the disease severity, the treatment properties and the frequency of motor complications in the patients with Parkinson's disease (PD) having and not having REM sleep behavior disorder (RBD). PATIENTS AND METHODS: Based on chart review, patients with Parkinson's disease whose bed partners have reported prominent motor activity while dreaming were identified. Standard questionnaires assessing the presence of RBD have been addressed to these patients and their informants. Obtained data fulfilled clinical diagnostic criteria of probable RBD in 35 patients (RBD group) with the mean age at symptom onset was 61.8 years. Of them 77% were men. Clinical features of these patients concerning Hoehn-Yahr stage of PD, the severity of PD according to the Unified Parkinson's disease rating scale (UPDRS), the mean dose and duration of levodopa (LD) therapy, the presence of motor complications were compared with those of gender and age at PD-onset matched 35 PD patients without RBD (NRBD group). RESULTS: The mean values of PD duration, Hoehn-Yahr stage and UPDRS scores did not differ between groups. The duration of LD therapy was significantly longer in RBD group in comparison to NRBD group (6.2 years versus 3.05 years, respectively, P<0.005) and also mean actual dose of LD was higher (460.3 mg/day versus 320.3 mg/day respectively, P<0.02). The dose and duration of dopamine agonists did not differ between groups. In RBD group, wearing-off phenomenon was significantly common (P<0.01), its duration was longer (P<0.005), and LD-related dyskinesias were more frequent (P<0.01). CONCLUSION: In the current study, when compared with NRBD group, the patients with RBD required higher doses of LD treatment at an earlier stage of PD which eventually led to motor complications. In these patients, dopaminergic treatment restored UPDRS scores, but did not prevent the occurrence of RBD. PMID- 15885390 TI - Manifestations of Neuro-Behcet's disease. Report of two cases and review of the literature. AB - Behcet's disease (BD) is a rare condition, more commonly seen in Turkey, the Middle East, the Mediterranean, and the Far Eastern countries. It is a multifaced disease involving the central nervous system (CNS), skin, joints, intestines and blood vessels. CNS involvement occurs in 10-25% of the patients with BD. We report two cases with Neuro-Behcet's disease. Both patients were females and they were in their second decade of life (early forties) when they suffered from relapsing retinitis and uveitis which led to blindness, despite treatment with steroids. They presented with clinical signs of brainstem syndrome, meningoencephalitis and the initial radiological investigations of our patients revealed encephalitic lesions in the brainstem. In the first case, an intracerebral abscess developed 4 years later, and it was evacuated under stereotactic guidance. The most common neuropathologic findings in Neuro-Behcet's disease are focal necrotic lesions in the brain. However, the association of Behcet's disease (BD) with cerebral abscess is very rare. There were only three reported cases of such. Ten years after the initial diagnosis of BD, our second patient suffered from subarachnoid hemorrhage due to a ruptured aneurysm of the superior cerebellar artery, which was then resected successfully. Intracranial aneurysm in patients with Behcet's disease has previously been documented in eight patients. To our knowledge, this is the only case of Neuro-Behcet's disease associated with an aneurysm of the superior cerebellar artery. PMID- 15885391 TI - Unilateral reduction of the cervical response in median SEP due to a vertebral fracture, unrecognized in plain films. AB - We report on a patient presenting with hypaesthesia in first, second and third finger of the right hand following a motorcycle accident. Conventional X-ray showed only a mild dislocation in C6/C7 segment. Cervical MRI in order to prove a root avulsion, was reported to be normal. Somatosensory evoked potentials (SEP) revealed a reduced amplitude of the cervical response on right median nerve stimulation. Needle-EMG showed a mild reduced recruitment pattern in triceps brachii muscle compatible with an anterior root lesion. Reviewing MRI, a signal loss in the course of C7 root was suspicious for an articular process fracture. This was proved in a CT scan. This case report emphasizes the topodiagnostic value of the reduced amplitude of the cervical potential of median nerve SEP and the importance of the CT in evaluating cervical spine fractures, as plain films frequent fail to do so. PMID- 15885392 TI - Giant nerve sheath tumour: report of six cases. AB - Giant nerve sheath tumour is a rare tumour originating from the nerve sheath. It differs from the conventional nerve sheath tumour only by the size these tumours can reach. There are two main type of tumours that occur in the nerve sheath which include neurofibroma and schwannoma. The current views are that schwannomas arise from the progenitor of the schwann cell. Whereas the neurofibroma series probably arise from a mesenchymal origin closer to a fibroblast. We report on six cases of nerve sheath tumour that occur in the spinal and paraspinal region that presented to us over a 5 year period. PMID- 15885393 TI - Surgical management of hydrocephalic dementia in Paget's disease of bone: the 6 year outcome of ventriculo-peritoneal shunting. AB - Paget's disease of bone is a chronic progressive skeletal disorder usually occurring in the long bones and skull of older adults and elderly persons. In the skull, softening of the skull base may lead to basilar impression and consequently obstruction of the cerebrospinal fluid through the basilar cisterns, resulting ventricular enlargement in association with gait difficulties, incontinence and dementia: a syndrome resembling normal pressure hydrocephalus. The optimal management of hydrocephalus associated with Paget's disease of the skull is not well documented and is still debated. We report a patient with hydrocephalic dementia linked to Paget's disease of the skull who showed marked sustained improvement in her neurological condition after ventriculo-peritoneal shunt insertion. We have now followed this patient for 6 years. Our experience supports the view that ventricular shunting is the procedure of choice for treating hydrocephalus linked to the Paget's disease of bone and is best carried out in the early stages of the illness. PMID- 15885394 TI - Calvarial metastases as clinical presentation of renal cell carcinoma: report of two cases and review of the literature. AB - Calvarium is a frequent target site of involvement for common neoplasms. Some cases of calvarial metastases have been reported in literature as secondary lesions from renal cell carcinoma (RCC), but only five cases have been described concerning calvarial mass as the first clinical presentation of this kind of tumor. In this report, we discuss the clinical aspects of two further cases we observed, in which the renal cell carcinoma was found thanks to the histological examination of a calvarial mass after surgery. We also briefly review the literature. PMID- 15885395 TI - Extra-axial ependymoma of posterior fossa extending to the Meckel's cave. AB - An extra-axial ependymoma extending from the left cerebellopontine corner to the Meckel's cave is reported. This lesion's clinical, radiological, and histological characteristics are presented. This tumor's infrequent extra-axial location, extension to the Meckel's cave and possible origin are discussed. PMID- 15885396 TI - Navigator system-assisted endoscopic fenestration of a symptomatic cyst in the septum pellucidum--technique and cases report. AB - Expanding cysts of the septum pellucidum are rare and frequently manifest as intermittent headaches. Although the technique of endoscopic fenestration has been used since 1999, only a limited number of cases have been reported. We have added the use of a navigator system to guide keyhole creation and endoscopic access. To provide experience in navigator endoscopic treatment of symptomatic cyst of septum pellucidum and long-term follow-up of the surgical result. Under the guidance of the navigator system, a burr hole was made and rigid endoscope was inserted into the lateral ventricle through a working sheath. With direct visualization, only one side of the lateral wall of the cyst was fenestrated. And a grasping basket was used to further dilate the perforated hole. Patient A, a 14 year-old male adolescent, had an acute onset of severe headache with increased intracranial pressure. Patient B was a 37-year-old woman with a diagnosis of medically intractable migraine. Both patients experienced dramatic symptomatic relief after surgery at 4.5- and 2-year follow-up exams, respectively. The technique of navigator-assisted endoscopic fenestration in the treatment of a symptomatic cyst of the septum pellucidum might be a safe and effective method. It achieved satisfactory results in our two patients. PMID- 15885397 TI - Jugular foramen syndrome caused by choleastatoma. AB - Unilateral paresis of 9th-11th cranial nerves together is defined as jugular foramen (Vernet's) syndrome. A cholesteatoma case that penetrated into intracranial area after eroding temporal bone and led to jugular foramen syndrome is presented here, since such a case has not been reported in the literature hitherto. A 46-year-old male patient was evaluated for hoarseness. It was learnt from his anamnesis that he had been operated for otitis media nearly 20 years ago. Uvula deviated towards right. Loss of gag reflex was remarkable on the left. Paresis was found on the left vocal cord. There was weakness in rotation of the head to the right. Computerized tomography (CT) scan revealed that the temporal bone lost its integrity and that there was an extra-axial hypodense mass neighboring pontocerebellum. Post-contrast magnetic resonance imaging showed that the mass, which showed thin, regular circular contrasting and which was hypointense in T1-weighted MR and hyperintense in T2-weighted MR, extended to the left jugular foramen. The mass was totally removed with left sub-occipital approach in the treatment. After the treatment, hoarseness, weakness in the rotation movement of the head and loss of gag reflex recovered totally, while deviation in the uvula was permanent. Cholesteatomas can extend to posterior fossa and cause jugular foramen syndrome. Early surgery is important to completely reverse the lost nerve functions in treatment. PMID- 15885398 TI - Cognitive recovery after delayed carbon monoxide encephalopathy. AB - We report on a patient with delayed carbon monoxide encephalopathy who presented with severe cognitive impairment associated with MRI findings of extensive demyelination of the cerebral white matter after a silent period of three weeks from acute intoxication. Despite the severe signs of structural and functional cerebral impairment in the sub-acute stage, the clinical course was favorable but for residual mild dysfunction of the frontal lobes. PMID- 15885399 TI - Corpus callosum changes following shunting for hydrocephalus: case report and review of the literature. AB - Diffuse callosal signal changes can rarely occur following successful shunting for obstructive hydrocephalus. We report on a patient who underwent ventriculoperitoneal shunting for aqueductal stenosis and required two revisions for neurological deterioration and presumed shunt malfunction. Extensive changes involving the corpus callosum, periventricular white matter and optic chiasm were noted after the first, and were prominent after second shunt revision. Thus, post shunting callosal changes may be associated with more extensive white matter disease, and may not imply wide pressure fluctuations. However, they need not prompt further intervention. PMID- 15885400 TI - Levels of polychlorinated dibenzo-p-dioxins and dibenzofurans in primipara breast milk from Taiwan: estimation of dioxins and furans intake for breastfed infants. AB - Postnatal exposure to dioxins in breastfed infants occurs mainly during breast feeding. The exposure to a substantial amount of endocrine disruptors in the early days of life may cause long-term health effects. Test subjects were healthy and primiparous mothers with a mean age of 28 (S.D. = 3.8) in 2001. The PCDD/F congeners were analyzed in the breast milk using gas chromatograph/high resolution mass spectrometry. The mean level of PCDD/Fs was 7.4 pg-WHO-TEQ/g lipid, which is significantly lower than the level found in individuals from other countries. The total PCDD WHO-TEQ levels in breast milk had a significant positive association with maternal age and a slightly negative association with perinatal BMI (body mass index of the period before and after the delivery). The estimated daily intake of 10.5 pg-WHO-TEQ/kg/day from individual breast milk was predicted for a breastfed infant at 6 months of age with proper assumption of 8 kg body weight, 854 milk per day of consumption, 95% of dioxin absorption rate, and linear decline of dioxin during lactation. Based on the lower WHO-TEQ levels in the breast milk, breast-feeding should still be encouraged and continued in Taiwan. PMID- 15885401 TI - Thermal reactivity of some nitro- and nitroso-compounds derived from 1,3,5,7 tetraazabicyclo[3.3.1]nonane at contamination by ammonium nitrate. AB - Thermal reactivity of 3,7-dinitro-1,3,5,7-tetraazabicyclo[3.3.1]nonane (DPT), 3,7 dinitroso-1,3,5,7-tetraazabicyclo[3.3.1]nonane (DNPT), 1,3,5-trinitroso-1,3,5 triazinane (TMTA or R-salt), 1,3,5-trinitro-1,3,5-triazinane (hexogen or RDX), 1,5-diacetyl-3,7-dinitro-1,3,5,7-tetrazocane (DADN), alpha-modification of the 1,3,5,7-tetranitro-1,3,5,7-tetrazocane (octogen or HMX) and of their mixtures with 2wt.% of ammonium nitrate (AN) has been examined by means of non-isothermal differential thermal analysis. The resulting data were analyzed according to the Kissinger method. The reactivity was expressed as the E(a)R(-1) slopes of the Kissinger relationship. A relatively high reactivity has been found with mixtures of DPT and DNPT with AN. Electronic charges q(N) at nitrogen atoms in molecules of the compounds studied were calculated by means of ab initio DFT B3LYP/6-31G** method. The relationships were confirmed between the slopes E(a)R(-1) and the q(N) values for the nitrogen atoms primarily undergoing reaction. On the basis of these relationships it is stated that the destabilizing effect of AN is due to acidolytic attack of nitric acid (resulting from dissociation of ammonium nitrate) at the nitrogen atoms with the most negative q(N) values in the molecules of the compounds studied. PMID- 15885402 TI - Experimentally validated 3-D simulation of shock waves generated by dense explosives in confined complex geometries. AB - Accidental blast wave generation and propagation in the surroundings poses severe threats for people and property. The prediction of overpressure maxima and its change with time at specified distances can lead to useful conclusions in quantitative risk analysis applications. In this paper, the use of a computational fluid dynamics (CFD) code CFX-5.6 on dense explosive detonation events is described. The work deals with the three-dimensional simulation of overpressure wave propagation generated by the detonation of a dense explosive within a small-scale branched tunnel. It also aids at validating the code against published experimental data as well as to study the way that the resulting shock wave propagates in a confined space configuration. Predicted overpressure histories were plotted and compared versus experimental measurements showing a reasonably good agreement. Overpressure maxima and corresponding times were found close to the measured ones confirming that CFDs may constitute a useful tool in explosion hazard assessment procedures. Moreover, it was found that blast wave propagates preserving supersonic speed along the tunnel accompanied by high overpressure levels, and indicating that space confinement favors the formation and maintenance of a shock rather than a weak pressure wave. PMID- 15885403 TI - A simple approach for determining detonation velocity of high explosive at any loading density. AB - A simple empirical relationship is introduced between detonation velocity at any loading density and chemical composition of high explosive as well as its gas phase heat of formation, which is calculated by group additivity rules. The present work may be applied to any explosive that contains the elements of carbon, hydrogen, nitrogen and oxygen with no difficulties. The new correlation can easily be applied for determining detonation velocity of explosives with loading densities less than 1g/cm3 as well as greater than 1g/cm3. Calculated detonation velocities by this procedure for both pure and explosive formulations show good agreement with respect to measured detonation velocity over a wide range of loading density. PMID- 15885404 TI - Flammability of gas mixtures. Part 1: fire potential. AB - International and European dangerous substances and dangerous goods regulations refer to the standard ISO 10156 (1996). This standard includes a test method and a calculation procedure for the determination of the flammability of gases and gas mixtures in air. The substance indices for the calculation, the so called "Tci values", which characterise the fire potential, are provided as well. These ISO Tci values are derived from explosion diagrams of older literature sources which do not take into account the test method and the test apparatus. However, since the explosion limits are influenced by apparatus parameters, the Tci values and lower explosion limits, given by the ISO tables, are inconsistent with those measured according to the test method of the same standard. In consequence, applying the ISO Tci values can result in wrong classifications. In this paper internationally accepted explosion limit test methods were evaluated and Tci values were derived from explosion diagrams. Therefore, an "open vessel" method with flame propagation criterion was favoured. These values were compared with the Tci values listed in ISO 10156. In most cases, significant deviations were found. A detailed study about the influence of inert gases on flammability is the objective of Part 2. PMID- 15885405 TI - Flammability of gas mixtures. Part 2: influence of inert gases. AB - Ternary systems, which contain flammable gas, inert gas and air, were studied in order to give the user an evaluation of the ISO 10156 calculation method for the flammability of gas mixtures. While in Part 1 of this article the fire potential of flammable gases was the focal point, the influence of inert gases on the flammability of gas mixtures was studied in Part 2. The inerting capacity of an inert gas is expressed by the dimensionless K value, the so-called "coefficient of nitrogen equivalency". The experimental determination of K values is demonstrated by using explosion diagrams. The objective of this study was to compare the estimated results, given by ISO 10156, with measurements of explosion ranges based on the German standard DIN 51649-1, given by CERN and CHEMSAFE. The comparison shows that ISO 10156, Table 1, supplies conservative K values, which can be regarded as safe in all cases. Nevertheless, in a number of cases ISO underestimates the inerting capacity, so that non-flammable gas mixtures are considered flammable. PMID- 15885406 TI - Removal of cadmium from wastewater using agricultural waste 'rice polish'. AB - A novel biosorbent rice polish has been successfully utilized for the removal of cadmium(II) from wastewater. The maximum removal of cadmium(II) was found to be 9.72 mg g(-1) at pH 8.6, initial Cd(II) concentration of 125 mg l(-1) and temperature of 20 degrees C. The effect of different parameters such as contact time, adsorbate concentration, pH of the medium and temperature were investigated. Dynamics of the sorption process were studied and the values of rate constant of adsorption, rate constant of intraparticle diffusion and mass transfer coefficient were calculated. Different thermodynamic parameters, viz., changes in standard free energy, enthalpy and entropy have also been evaluated and it has been found that the reaction was spontaneous and exothermic in nature. The applicability of Langmuir isotherm showed monolayer coverage of the adsorbate on the surface of adsorbents. A generalised empirical model was proposed for the kinetics at different initial concentrations. The data were subjected to multiple regression analysis and a model was developed to predict the removal of Cd(II) from wastewater. PMID- 15885407 TI - Removal of arsenic from water by zero-valent iron. AB - Batch and column experiments were conducted to investigate the effect of dissolved oxygen (DO) and pH on arsenic removal with zero-valent iron [Fe(0)]. Arsenic removal was dramatically affected by the DO content and the pH of the solution. Under oxic conditions, arsenate [As(V)] removal by Fe(0) filings was faster than arsenite [As(III)]. Greater than 99.8% of the As(V) was removed whereas 82.6% of the As(III) was removed at pH 6 after 9h of mixing. When the solution was purged with nitrogen gas to remove DO, less than 10% of the As(III) and As(V) was removed. High DO content and low solution pH also increased the rate of iron corrosion. The removal of arsenic by Fe(0) was attributed to adsorption by iron hydroxides generated from the oxic corrosion of Fe(0). The column results indicated that a filtration system consisting of an iron column and a sand filter could be used for treatment of arsenic in drinking water. PMID- 15885408 TI - Kinetic study of the immobilization of galvanic sludge in clay-based matrix. AB - The viability of inertization of galvanic wastes through their incorporation in clay-based materials, such as common formulations for tiles and bricks, is here studied by determining the leaching kinetics in different media. Metals immobilization is assured by firing at reasonably high temperatures, since intimately contact and/or reaction between residue and clay particles is promoted but also due to formation of insoluble metal oxides that rest unreactive towards clay grains. For most metals, leaching rate follows a zero-order kinetic law, with values between 0.001 to 0.1 mg/(g day cm2). Leaching velocity tends to increase with rising atomic numbers: Zn < Cu < Ni < Cr. These values depend exponentially on the relative sludge content. PMID- 15885409 TI - Enrichment/separation of cadmium(II) and lead(II) in environmental samples by solid phase extraction. AB - A preconcentration/separation procedure is presented for the solid phase extraction of trace cadmium and lead ions as their 1-(2-pyridylazo) 2-naphthol (PAN) chelates in environmental samples on Chromosorb-106 resin, prior to cadmium and lead determinations by atomic absorption spectrometry. The preconcentration procedure was optimized by using model solutions containing cadmium and lead ions. The influences of pH of the model solutions, amounts of PAN, eluent type and volume etc. were investigated. Also the effects of the matrix constituents of the samples were also examined. Separation of cadmium and lead from real samples was achieved quantitatively. The procedure presented was checked with the analysis of microwave-digested standard reference materials (IAEA-336 Lichen and SRM 1515 Apple leaves). The preconcentration procedure was applied for the lead and cadmium contents of the natural water samples, some salts with satisfactory results (recoveries >95%, relative standard deviations <8%). PMID- 15885410 TI - A hybrid liquid-phase precipitation (LPP) process in conjunction with membrane distillation (MD) for the treatment of the INEEL sodium-bearing liquid waste. AB - A novel hybrid system combining liquid-phase precipitation (LPP) and membrane distillation (MD) is integrated for the treatment of the INEEL sodium-bearing liquid waste. The integrated system provides a "full separation" approach that consists of three main processing stages. The first stage is focused on the separation and recovery of nitric acid from the bulk of the waste stream using vacuum membrane distillation (VMD). In the second stage, polyvalent cations (mainly TRU elements and their fission products except cesium along with aluminum and other toxic metals) are separated from the bulk of monovalent anions and cations (dominantly sodium nitrate) by a front-end LPP. In the third stage, MD is used first to concentrate sodium nitrate to near saturation followed by a rear end LPP to precipitate and separate sodium nitrate along with the remaining minor species from the bulk of the aqueous phase. The LPP-MD hybrid system uses a small amount of an additive and energy to carry out the treatment, addresses multiple critical species, extracts an economic value from some of waste species, generates minimal waste with suitable disposal paths, and offers rapid deployment. As such, the LPP-MD could be a valuable tool for multiple needs across the DOE complex where no effective or economic alternatives are available. PMID- 15885411 TI - Bioremediation of Cr(VI) in contaminated soils. AB - Ex situ treatment of hexavalent chromium (Cr(VI)) contaminated soil using a bioreactor-biosorption system was evaluated as a novel remediation alternative. Leaching of Cr(VI) from the contaminated soil using various eluents showed that desorption was strongly affected by the solution pH. The leaching process was accelerated at alkaline conditions (pH 9). Though, desorption potential of ethylene diamine tetra acetic acid (EDTA) was the maximum among various eluents tried, molasses (5 g/L) could also elute 72% of Cr(VI). Cr(VI) reduction studies were carried out under aerobic and facultative anaerobic conditions using the bacterial isolates from contaminated soil. Cr(VI) reduction was moderately higher in aerobic conditions than in facultative anaerobic conditions. The effect of various electron donors on Cr(VI) reduction was also investigated. Among five electron donors screened, peptone (10 g/L) showed maximum Cr(VI) reduction followed by molasses (10 g/L). The time required for complete Cr(VI) reduction was increased with increase in the initial Cr(VI) concentration. However, specific Cr(VI) reduction was increased with increase in initial Cr(VI) concentration. Sulfates and nitrates did not compete with Cr(VI) for accepting the electrons. A bioreactor was developed for the detoxification of Cr(VI). Above 80% of Cr(VI) reduction was achieved in the bioreactor with an initial Cr(VI) concentration of 50 mg/L at an HRT of 8 h. An adsorption column was developed using Ganoderm lucidum (a wood rooting fungus) as the adsorbent for the removal of trivalent chromium (Cr(III)) and excess electron donor from the effluent of the bioreactor. The specific Cr(III) adsorption capacity of G. lucidum in the column was 576 mg/g. The new biosystem seems to be a promising alternative for the ex situ bioremediation of Cr(VI) contaminated soils. PMID- 15885412 TI - A study of kinetic modelling and reaction pathway of 2,4-dichlorophenol transformation by photo-fenton-like oxidation. AB - The degradation of 2,4-dichlorophenol (2,4-DCP) by the photo-Fenton-like (Fe3+/H2O2/UVC) process under various reaction conditions was investigated. It was interesting to find that the reaction kinetics of 2,4-DCP in Fe3+/H2O2/UVC systems varied depending on the initial [Fe3+] concentration. A pseudo first order kinetic and a non-conventional kinetic were discovered at low and higher [Fe3+] concentrations, respectively. A model was used to simulate the non conventional kinetic process, where two character parameters (the initial decay rate and the final decay fraction) were found to be critical in determining the process. The two parameters successfully quantify the photo-Fenton-like oxidation under different concentrations of [Fe3+] and [H2O2] and the corresponding ratios of Fe(III)/H2O2. The reaction intermediates were identified by an LC/MS analysis and a reaction mechanism was proposed. PMID- 15885413 TI - Pre-ozonation coupled with UV/H2O2 process for the decolorization and mineralization of cotton dyeing effluent and synthesized C.I. Direct Black 22 wastewater. AB - The decolorization and mineralization of cotton dyeing effluent containing C.I. Acid Black 22 as well as synthesized C.I. Acid Black 22 wastewater by means of advanced oxidation processes (AOPs), such as UV/H2O2, O3 and pre-ozonation coupled with UV/H2O2 processes, were evaluated in this study. It was observed that the UV/H2O2 process took longer retention time than ozonation for color removal of dye bath effluent. Reversely, the total organic carbon (TOC) removal showed different phenomena that ozonation and UV/H2O2 process obtained 33 and 90% of removal efficiency for 160 min of retention time, respectively. Additionally, laboratory synthesized dye wastewater was substantially more efficient in the decolorization process than dye bath effluent. Therefore, in this work, pre ozonation coupled with UV/H2O2 process was employed to enhance the reduction of both color and TOC in dye bath effluent at the same time. At the same time, the retention time demand was reduced to less than 115 min for 90% removal of TOC and color by this combined process. PMID- 15885414 TI - Effect of adsorbent concentration to the adsorption of phenol on hexadecyl trimethyl ammonium-bentonite. AB - In this work, it is intended to study the effect of adsorbent concentration on the adsorption of phenol by hexadecyl trimethyl ammonium-bentonite. The experiments were conducted in two groups. The adsorption of hexadecyl trimethyl ammonium bromide (HDTMA) on bentonite was studied in the first group of experiments. It was observed that the all HDTMA was adsorbed by the bentonite, even when the amount used exceeded 100% of cation exchange capacity (CEC). After the modification of bentonite by using HDTMA in an amount equivalent to 100% of CEC, the adsorption experiments were performed at five different adsorbent concentrations ranging from 2 to 10 g/L. A type V isotherm and a non-linear increase in percent removal with adsorbent concentration were observed. The observation of the non-linear relation between the percent removal and adsorbent concentration was attributed to the effect of intra particle interactions and it was represented by a second order polynomial. Several adsorption isotherm equations were applied to the experimental data. Although, the Freundlich equation fitted fairly well, it failed to represent the plateau and the second region that appeared in the isotherm. Therefore, an equation giving the equilibrium concentration as a function of initial and adsorbent concentrations was suggested. PMID- 15885415 TI - Physical and chemical characterisation of crude meat and bone meal combustion residue: "waste or raw material?". AB - As a result of the recent bovine spongiform encephalopathy (BSE) crisis in the European beef industry, the use of animal by-product is now severely controlled. Meat and bone meal (MBM) production can no longer be used to feed cattle and must be safely disposed of or transformed. Main disposal option is incineration, producing huge amounts of ashes the valorisation of which becomes a major concern. The aim of this work is to characterise MBM combustion residue in order to evaluate their physical and chemical properties to propose new valorisation avenues. The thermal behaviour of crude meat and bone meal was followed by thermogravimetric analysis (TGA) and (24 wt.%) inorganic residue was collected. The resulting ashes were characterised by powder X-ray diffraction (XRD), particle size distribution, specific surface area (BET), scanning electron microscopy (SEM) couple with energy disperse X-ray analysis (EDX). Elemental analysis revealed the presence of chloride, sodium, potassium, magnesium with high level of phosphate (56 wt.%) and calcium (31 wt.%), two major constituents of bone, mainly as a mixture of Ca10(PO4)6(OH)2 and Ca3(PO4)2 phases. The impact of combustion temperature (from 550 to 1000 degrees C) on the constitution of ashes was followed by TGA, XRD and specific surface measurements. We observed a strong decrease of surface area for the ashes with crystallisation of calcium phosphates phases without major changes of chemical composition. PMID- 15885416 TI - Leaching of APC residues from secondary Pb metallurgy using single extraction tests: the mineralogical and the geochemical approach. AB - Two air-pollution-control (APC) residues--one from flue gas cooling with alkaline water and one from deionized water cooling--from secondary lead metallurgy were submitted to two different standardized short-term leaching protocols: US EPA toxicity characteristic leaching procedure (TCLP) and static leaching according to Czech/European norm EN 12457-2. The experimental procedure was coupled with detailed mineralogical investigation of the solid material (SEM, XRPD) and speciation-solubility calculations using the PHREEQC-2 geochemical code. Both types of residues were considered as hazardous materials exhibiting substantial leaching of Pb (up to 7130 mg/l) and other inorganic contaminants. However, the APC residue produced by flue gas cooling with alkaline water (sample B) exhibits more favourable leaching and environmental characteristics than that produced by simple deionised water cooling (sample A). At pH < 5, primary caracolite (Na3Pb2(SO4)3Cl) and potassium lead chloride (KCl.2PbCl2) are completely or partially dissolved and transformed to residual anglesite (PbSO4), cotunnite (PbCl2) and laurionite (Pb(OH)Cl). At pH 5-6, anglesite is still the principal residual product, whereas at pH > 6, phosgenite (PbCl2.PbCO3) became the dominant secondary phase. The results are consistent with the mineralogical and geochemical studies focused on acidic forest soils highly polluted by smelter emissions, where anglesite, as a unique Pb-bearing phase, has been detected. From the technological point of view, the mixing of APC residue with alkaline water, followed by an increase in the suspension pH and equilibration with atmospheric CO2, may be used to ensure the precipitation of less soluble Pb carbonates, which are more easily recycled in the Pb recovery process in the metallurgical plant. PMID- 15885417 TI - Kinetics of degradation of sulphur mustard on impregnated carbons. AB - Kinetics of degradation of sulphur mustard (HD) on the surface of NaOH/CrO3/C, NaOH/CrO3/EDA/C and RuCl3/C systems has been examined by using gas chromatography technique by extracting and analyzing the residual HD periodically. The carbons were prepared by impregnating activated carbon with 4% sodium hydroxide plus 3% Cr(VI) as CrO3 with and without 5% ethylene diamine (EDA) and 5% ruthenium chloride by using their aqueous solutions. Obtained carbons were characterized for surface area analysis by BET conventional method. Kinetic plots reveal that the observed reactions are fast at the initial stages, slow at the later stages and progress to a steady state indicating the first order behavior. Effect of moisture on kinetic rate is also observed. In the case of NaOH/CrO3/C system the rate constant is decreased from 13.36 to 5.53 x 10(-2) h(-1) and half life is increased from 5.2 to 12.54 h while moisture content is increased from 1.9% to 11.2%. Whereas, the rate constant of HD degradation reaction is decreased from 10.4 to 4.14 x 10(-2) h(-1) and half life is increased from 6.7 to 16.72 h while moisture content is increased from 2.1% to 10.8% on NaOH/CrO3/EDA/C. Reaction on RuCl3/C system also behaves in the similar manner. Extracted reaction products were characterized by GC/MS and it is found that on NaOH/CrO3/C, HD degrades to hemisulphur mustard, thiodiglycol and 1,4-oxathiane. Whereas, on NaOH/CrO3/EDA/C, HD is degraded to 1,4-thiazane and it is degraded to divinyl sulphone on RuCl3/C. All these investigations reveal that above mentioned carbons can be used in nuclear, biological and chemical (NBC) filtration systems for protection against sulphur mustard. PMID- 15885418 TI - Biosorption of Cr(III) ions by eggshells. AB - The paper presents results of studies carried out on sorption of Cr(III) ions from aqueous solutions by eggshells as a low-cost sorbent. It was found that crushed eggshells possess relatively high sorption capacity, when comparing with other sorbents, that was evaluated as 21-160 mg/g. The effect of process parameters: pH, temperature, initial concentration of Cr(III) ions on the process kinetics was studied. It was found that the equilibrium of the process was reached after 60 min. Also equilibrium studies were performed: the effect of sorbent concentration and equilibrium Cr(III) concentration was studied. The maximum experimentally determined sorption capacity 160 mg/g was obtained at low sorbent concentration at 20 degrees C and pH 5. It was found that sorption capacity increased with the increase of Cr(III) concentration, temperature and sorbent concentration. Mathematical models describing kinetics and equilibrium of sorption were proposed. The process kinetics was described with pseudo-second order pattern and equilibrium was described with Langmuir-type equation, and the influence of sorbate concentration, with an empirical dependence. The models were positively verified. Eggshells were able to remove the concentration of Cr(III) ions below the acceptable level, i.e. at 40 degrees C, at the initial concentration of metal ions 100 mg/kg, at sorbent concentration 15 g/l. PMID- 15885419 TI - The effect of ash and filter media characteristics on particle filtration efficiency in fluidized bed. AB - The phenomenon of filtering particles by a fluidized bed is complex and the parameters that affect the control efficiency of filtration have not yet been clarified. The major objective of the study focuses on the effect of characteristics of ash and filter media on filtration efficiency in a fluidized bed. The performance of the fluidized bed for removal of particles in flue gas at various fluidized operating conditions, and then the mechanisms of collecting particles were studied. The evaluated parameters included (1) various ashes (coal ash and incinerator ash); (2) bed material size; (3) operating gas velocity; and (4) bed temperature. The results indicate that the removal efficiency of coal ash increases initially with gas velocity, then decreases gradually as velocity exceeds some specific value. Furthermore, the removal of coal ash enhance with silica sand size decreasing. When the fluidized bed is operated at high temperature, diffusion is a more important mechanism than at room temperature especially for small particles. Although the inertial impaction is the main collection mechanism, the "bounce off" effect when the particles collide with the bed material could reduce the removal efficiency significantly. Because of layer inversion in fluidized bed, the removal efficiency of incinerator ash is decreased with increasing of gas velocity. PMID- 15885420 TI - Optimization of phosphorus removal from secondary effluent using simplex method in Tianjin, China. AB - Enhanced concentrations of phosphorus entering the aquatic systems have been linked with eutrophication and its associated problems. Jar tests were applied to a secondary effluent in order to determine optimal conditions for coagulation. The coagulants studied were ferric sulfate [Fe2(SO4)3], aluminum sulfate [Al2(SO4)3.18H2O] and polyaluminum chloride (PAC). The experiments were carried out using simplex research technique to determine the optimum conditions of these coagulants for phosphorus removal. For each coagulant examined, 18-25 experiments were carried out until a maximum removal was observed through the experimental procession. Increases of greater than 30% were found for the removal efficiencies of these three coagulants over the course of the experiments. Good removal efficiencies averaging at least 87.25% were obtained through the simplex procession. The orthophosphate removal efficiency was higher than that of total phosphorus for each coagulant. The maximum removal efficiency of total phosphorus obtained from the optimization procedures for ferric sulfate, aluminum sulfate and polyaluminum chloride was 87.3, 95.6 and 94.0%, respectively, and the minimum total phosphorus residual was 0.35, 0.12 and 0.16 mg/L, respectively. PMID- 15885421 TI - The use of non-uniform electrokinetics to enhance in situ bioremediation of phenol-contaminated soil. AB - In situ bioremediation is an attractive and often cost-effective technology for the cleanup of organics-contaminated sites, but it often requires extended treatment time under field conditions. This study explored the feasibility of using non-uniform electrokinetic transport processes to enhance in situ bioremediation. A bench-scale non-uniform electrokinetic system with periodic polarity-reversal was developed for this purpose, and tested by using a sandy loam spiked with phenol as a model organic pollutant. The results demonstrated that non-uniform electrokinetic processes could accelerate the movement and in situ biodegradation of phenol in the soil. Bidirectional operation enhanced the phenol biodegradation more effectively than unidirectional operation. At the same time, a smaller polarity-reversing interval induced a higher and more uniform removal of phenol from the soil. The results also showed that reversing the polarity of electric field applied could maintain the soil pH and moisture, but it increased the consumption of electricity. PMID- 15885422 TI - Leaching behaviour of a galvanic sludge in sulphuric acid and ammoniacal media. AB - Leaching studies of a sludge produced by the physico-chemical treatment of wastewaters generated by a Ni/Cr plating plant were carried out in both sulphuric acid and ammoniacal media aiming to decide which of them would be the best treatment for this kind of waste material. The dissolution behaviour of some metals (Cu, Ni, Cr and Zn) was studied in order to assure the best metal recovery conditions in subsequent processes by the use of some separation methods such as solvent extraction and precipitation techniques. Therefore, the study here presented deals with the first chemical stage of an integrated treatment process. For the sulphuric acid leaching, maximal conversions obtained were 88.6% Cu, 98.0% Ni and 99.2% Zn for the following experimental conditions: a 100 g L(-1) acid concentration, a 5:1 liquid-to-solid ratio (L/S), a particle size less than 1 mm, a digestion time of 1h, a stirring speed of 700 rpm (all at room temperature and under atmospheric pressure). As expected, no selectivity was achieved for the sulphuric acid leaching, despite this option yielding much higher metal ion dissolution when compared with that reached by ammoniacal leaching. The use of this latter medium allowed the extraction of Cu and Ni without Cr species, but rates of conversion were only about 70% for Cu and 50% for Ni, much lower than those obtained for sulphuric acid leaching. PMID- 15885423 TI - Interactions of aqueous Cu2+, Zn2+ and Pb2+ ions with crushed concrete fines. AB - The crushing of reclaimed concrete-based demolition waste to produce recycled aggregate gives rise to a large volume of cement-rich fine material for which market development would be beneficial. It was envisaged that this fine fraction may prove to be an effective sorbent for aqueous heavy metal species by virtue of its ion exchangeable phases and high pH. A batch sorption study confirmed that crushed concrete, in the particle size range 1-2 mm, successfully excluded Cu2+ (35 mg g(-1)), Zn2+ (33 mg g(-1)) and Pb2+ (37 mg g(-1)) from aqueous media. Subsequent distilled water leaching of the metal-laden concrete particles indicated that 1.9, 0.9 and 0.2% of the bound metals, Cu2+, Zn2+ and Pb2+, respectively, were readily soluble. Scanning electron microscopy revealed that the removal of Cu2+ and Zn2+ arose from surface precipitation reactions, whereas, the principal mechanism of uptake of Pb2+ was found to be by diffusion into the cement matrix. The metal ion removal efficiency of crushed concrete fines is compared with those of other low cost sorbents and potential applications which may exploit this sorptive property are also discussed. PMID- 15885424 TI - Oxidation of Cr(III) in tannery sludge to Cr(VI): field observations and theoretical assessment. AB - Sludge, soil and leachate samples collected from a chromium-contaminated tannery waste dumping site in Kanpur, India, were found to contain considerable amounts of Cr(VI), despite the fresh tannery sludge containing little or no Cr(VI). Literature reports suggested that dry Cr(III) precipitates could be converted to Cr(VI) when heated in the presence of oxygen. Also, Cr(III) in aqueous phase could be oxidized through interaction with manganese dioxide (MnO2) surface to Cr(VI). Measurement of manganese in the sludge samples collected from the site showed concentrations up to 0.6 mg/g. Based on equilibrium calculations, it was determined that both dry phase Cr(III) oxidation by atmospheric oxygen and aqueous phase Cr(III) oxidation by MnO2 surface were thermodynamically feasible. It was further suggested that in aqueous phase, manganese may act effectively as an electron transporter between Cr(III) and dissolved oxygen during Cr(III) oxidation, leading to regeneration of MnO2 solid phase. Further, as dissolved Cr(III) is oxidized, dissolution of Cr(OH3) will take place to maintain the equilibrium between the dissolved and solid phases of Cr(III). In the pH range of 3-10, and at oxygen partial pressure (P(O2)) of 10(-6) atm or higher, equilibrium conditions stipulate nearly complete conversion of Cr(III) to Cr(VI). At P(O2) of 10(-20) atm or lower, very little Cr(VI) is expected to be present under equilibrium conditions. In the intermediate P(O2) regions, incomplete dissolution of the Cr(OH3) solid phase and only partial conversion of chromium from +3 to the +6 oxidation state is expected, especially at lower pH values. PMID- 15885425 TI - Determination of solid waste sorption capacity for selected heavy metals in landfills. AB - The adsorption process is largely a surface-action phenomenon. In this study, sorption capacities for heavy metals on a solid waste matrix were investigated. Five heavy metals (iron, copper, zinc, nickel and cadmium) were chosen because of their availability in any landfill site. The conditions during all the experimental runs were pH 7.0, temperature 32 degrees C and suppressed microbial degradation. For adsorption isotherm (Freundlich and Langmuir) calculations, fixed quantities of heavy metal ions were mixed with variable quantities of solid waste. The ratio of mass of adsorbate per unit mass of adsorbent was changed five times, by changing only the adsorbent amount. The results showed that the time required to reach equilibrium varied from metal to metal but all reached equilibrium within the first 32 h. The relative potential of sorption of the individual metals and mixed metals on the solid waste matrix is Fe > Zn > Cu > Ni > Cd. The sorption capacity of domestic solid waste matrix for heavy metals is quite significant and this property might prove helpful for the in situ removal of heavy metals in landfill operation. PMID- 15885426 TI - Adsorption of phenolic compounds from aqueous solution onto a macroporous polymer and its aminated derivative: isotherm analysis. AB - Adsorption of phenolic compounds from aqueous solution to a macroporous polymeric adsorbent (CHA-111), its animated derivative (MCH-111) and a reference weakly anion exchanger (ND-900) was studied. Experimental results indicated that amino functional groups on the polymeric matrix play an important role in phenol adsorption by MCH-111 and ND-900, which was attributed to the formation of hydrogen bonding between the phenol molecule and the amino group on the polymeric matrix. The semi-empirical Freundlich isotherm equation and its reduced form were employed to interpret the adsorption behavior. A site energy distribution model based on the Polanyi adsorption potential theory can elucidate the adsorption mechanism reasonably. PMID- 15885427 TI - Removal of nickel from aqueous solutions by sawdust. AB - The main parameters influencing Nickel (II) metal sorption on maple sawdust were: initial metal ion concentration, amount of adsorbent, and pH value of solution. The maximum percent metal removal was attained after about 1h. The greatest increase in the rate of adsorption of metal ions on sawdust was observed for pH changes from 2 to 5. An empirical relationship has been obtained to predict the percentage Nickel (II) removal at any time for known values of sorbent and initial sorbate concentration. The experimental results provided evidence for chelation ion exchange as the major adsorption mechanisms for binding metal ions to the sawdust. The adsorbent can be effectively regenerated using 0.1 M strong acid and reused. PMID- 15885428 TI - Removal of cationic dyes from aqueous solution by adsorption on peanut hull. AB - The potential feasibility of peanut hull particle for removal of three cationic dyes (methylene blue, brilliant cresyl blue and neutral red) from aqueous solution was investigated. The effects of various experimental parameters were examined and optimal experimental conditions were decided. Above the value of initial pH 4, three dyes studied could be removed effectively. The isothermal data fitted the Langmuir model or Freundlich model. The adsorption processes followed the pseudo-first-order rate kinetics. The results in this study indicated that peanut hull was an attractive candidate for removing cationic dyes from dye wastewater. PMID- 15885429 TI - Modification of MSW fly ash by anionic chelating surfactant. AB - This paper elucidates a study on the re-utilization and stabilization of municipal solid waste (MSW) fly ash in producing a high value-added product by the surface modification of anionic chelating surfactant on the particles. After modification, MSW fly ash can be expected using as a filler of ultra-high molecular weight polymers. The effects of anionic chelating surfactants (ACS) on surface modification of MSW fly ash and fixing capacity for heavy metals were explored. Meanwhile, the interaction mechanism between surfactants and MSW fly ash was suggested. The results showed that anionic chelating surfactants can be used to effectively modify MSW fly ash particles and achieve a high active ratio. At the same time, they also exhibited a strong fixing capacity for heavy metals. Of the two modified MSW fly ash, ED3A-modified MSW fly ash has a much higher active ratio than MAP-modified MSW fly ash at over 95%, although its fixing capacity for heavy metals was a shade lower than MAP-modified MSW fly ash. PMID- 15885430 TI - A method of dynamic chromatic aberration correction in low-voltage scanning electron microscopes. AB - A time-of-flight concept that dynamically corrects for chromatic aberration effects in scanning electron microscopes (SEMs) is presented. The method is predicted to reduce the microscope's chromatic aberration by an order of magnitude. The scheme should significantly improve the spatial resolution of low voltage scanning electron microscopes (LVSEMs). The dynamic means of correcting for chromatic aberration also allows for the possibility of obtaining high image resolution from electron guns that have relatively large energy spreads. PMID- 15885431 TI - Use of surface affinity enrichment and cryo-embedding to prepare in vitro reconstituted mitotic chromosomes for EM tomography. AB - We present a novel preparation method for studies of in vitro reconstituted mitotic chromosomes from Xenopus laevis egg extracts. This method involves a gentle adsorption of chromosomes from the extracts using surface affinity enrichment, followed by plunge freezing, freeze-substitution and cryo-embedding before examination by EM tomography. For comparison, chromosomes were also prepared by a conventional method, which included immobilization of chromosomes in agarose and a room-temperature dehydration/embedding protocol. Three dimensional reconstructions showed that samples prepared with the new method have a greater interconnectivity of sub-structural features and a higher compaction ratio together with an apparently less perturbed chromatin structure than those prepared using the conventional approach. The implications of the new method for the preparation of other difficult samples and additional application possibilities are discussed. PMID- 15885432 TI - Refinements in the collection of energy filtered diffraction patterns from disordered materials. AB - In this paper a method for collecting electron diffraction patterns using a Gatan imaging filter is presented. The method enables high-quality diffraction data to be measured at scattering angles comparable to those that can be obtained using X ray and neutron diffraction. In addition, the method offers the capability for examining small regions of sample in, for example, thin films and nano structures. Using X-ray, neutron and electron diffraction data collected from the same sample, we demonstrate quantitative agreement between all three. We also present a novel method for obtaining the single scattering contribution to the total diffracted intensity by collecting data at various electron wavelengths. This approach allows pair distribution functions to be determined from electron diffraction in cases where there exists significant multiple scattering. PMID- 15885433 TI - IMAGE-WARP: a real-space restoration method for high-resolution STEM images using quantitative HRTEM analysis. AB - We have developed a new method for processing distorted high-resolution scanning transmission electron microscopy (STEM) images. The method is based on finding the displaced vertices in the experimental STEM image and warping to geometrically correct reference grid of the object. As a reference grid for warping a structural model obtained using a high-resolution transmission electron microscopy (HRTEM) analysis of the area of interest is utilised. Combined with quantitative HRTEM analysis the IMAGE-WARP method provides a real-space restoration of high-resolution high-angle annular dark-field (HAADF) STEM images without affecting the original Z-contrast information. The method can be applied to extract valuable compositional atomic-column data from any HAADF-STEM image of any kind of bulk crystals with local occupancy or chemistry fluctuations, stacking faults, special grain boundaries or interfaces, for which we have an available structural model. After the warping, distortion-corrected images can be further enhanced using conventional image-filtering techniques, and finally quantified with HAADF-STEM image simulations. The applicability of the IMAGE-WARP method was illustrated using experimental HAADF-STEM images of a strontium titanate crystal disrupted with a Ruddlesden-Popper-type antiphase boundary. PMID- 15885434 TI - Spline-based image-to-volume registration for three-dimensional electron microscopy. AB - This paper presents an algorithm based on a continuous framework for a posteriori angular and translational assignment in three-dimensional electron microscopy (3DEM) of single particles. Our algorithm can be used advantageously to refine the assignment of standard quantized-parameter methods by registering the images to a reference 3D particle model. We achieve the registration by employing a gradient-based iterative minimization of a least-squares measure of dissimilarity between an image and a projection of the volume in the Fourier transform (FT) domain. We compute the FT of the projection using the central-slice theorem (CST). To compute the gradient accurately, we take advantage of a cubic B-spline model of the data in the frequency domain. To improve the robustness of the algorithm, we weight the cost function in the FT domain and apply a "mixed" strategy for the assignment based on the minimum value of the cost function at registration for several different initializations. We validate our algorithm in a fully controlled simulation environment. We show that the mixed strategy improves the assignment accuracy; on our data, the quality of the angular and translational assignment was better than 2 voxel (i.e., 6.54 angstroms). We also test the performance of our algorithm on real EM data. We conclude that our algorithm outperforms a standard projection-matching refinement in terms of both consistency of 3D reconstructions and speed. PMID- 15885435 TI - Scripting-customized microscopy tools for Digital Micrograph. AB - Software is an integral part of all electron microscopy systems, encompassing hardware control, data acquisition and processing. It is unlikely that any one software system will meet all the requirements of experienced users. However, if the software supports custom scripting, then users are well placed to address any shortcomings by writing their own software. In this paper, we highlight the scripting capability within Gatan Inc.'s Digital Micrograph (DM) software, a widely used program for TEM imaging and EELS spectroscopy. We show how scripting can greatly extend the capabilities of the DM software, in tasks ranging in complexity from simple image manipulation through to full-blown microscope/imaging filter control and data acquisition. Scripting enables customized software tools to be developed to meet individual experimental needs, something which no software manufacturer could ever hope to do on a commercial basis. In essence, scripting allows the microscopist to drive the software rather than the software drive the microscopist. To foster an increased awareness and interest in DM scripting we have developed a web-based archive for DM scripts, which is freely accessible via the internet. PMID- 15885436 TI - Enzyme immunoassay of progesterone in the feces from beef cattle to monitor the ovarian cycle. AB - The present study was undertaken to measure fecal progesterone concentration of beef cattle using antibody against authentic progesterone and to examine whether this method can monitor the ovarian cycle in beef cattle. Rectal fecal samples collected from 14 beef cattle were mixed with 6 ml of 100% methanol and shaken for 15 min. After centrifugation, supernatant was extracted with petroleum ether followed by an enzyme immunoassay (EIA) for progesterone. Specificity of the assay was examined by HPLC separation of fecal solution followed by the EIA in each fraction. The present assay identified only progesterone but not other metabolites in the feces sample that was extracted with petroleum ether. Sensitivity of the assay was estimated to be 0.0055 ng/ml (0.11 ng/g). Coefficient variations of intra- and inter-assay were 9.6-10.9% and 10.8-16.6%, respectively. Recovery rates ranged between 73 and 84%. Patterns in the fecal progesterone concentrations during the ovarian cycle were almost parallel to the plasma concentrations. A significant positive correlation was established between the fecal and plasma progesterone concentrations in individual animal (r=0.59 0.84, P<0.001, n=10) as well as pooled data (r=0.70, P<0.001, n=65). Fecal progesterone concentrations of day 0 (showing the nadir of concentration) of the ovarian cycle were less than 50 ng/g, which increased significantly toward day 9 (P<0.01). From days 14 to 18, there was significant reduction of fecal progesterone concentration (P<0.01). Ovarian cycles had at least 48 ng/g (mean=74 ng/g) of difference between minimum and maximum fecal progesterone concentrations. All cattle at days 9, 11 and 14 had higher fecal progesterone concentrations by more than 20 ng/g compared with day 0. These results suggest that the present EIA is suitable to measure the progesterone in cattle feces and can monitor ovarian cycle. PMID- 15885437 TI - Restoration patterns for luteinising hormone and ovarian function following treatment with GnRH agonist implants (deslorelin) for 7, 14 or 21 days in cycling dairy cows. AB - Continuous GnRH agonist treatment of cows results in downregulation of GnRH responsiveness and a state of induced anoestrus. Inducing anoestrus in a precisely controlled manner could have several potential applications in dairy herd management. However, relatively little is known regarding the processes involved in restoring reproductive normality following an induced anoestrus. This study describes an experiment that was conducted to examine patterns of recovery of LH release and follicle growth in non-lactating Holstein cows immediately following cessation of treatment for 7, 14 or 21 days with a deslorelin implant. Oestrus cycles were synchronized at 7 days intervals and a deslorelin implant inserted in every cow 13 days after detected oestrus so that a group had implants for either 21 days (n = 9), 14 days (n = 10) or 7 days (n = 9). On the day of implant removal every ovarian follicle greater than 4 mm in diameter was ablated using ultrasound guided vacuum needle aspiration in an attempt to standardize follicle sizes. Daily ovarian ultrasound examinations were performed on each cow until 35 days after implant removal and again at 45, 59 and 74 days. A subgroup of four cows randomly selected from each treatment group had frequent serial blood samples collected over 8 h at 4 and 10 days after implant removal for LH profiling. There was no significant effect of treatment duration on any LH parameter and results were pooled. Mean LH pulse amplitude increased by 67% between 4 and 10 days after implant removal (0.34 ng/ml versus 0.57 ng/ml; 4 days versus 10 days post-implant, P < 0.001). Mean pulse frequency remained unchanged between the two samplings (5.9 pulses versus 6.9 pulses per 8 h; 4 days versus 10 days post-implant, P > 0.1). Smoothed mean LH concentrations were unaffected by treatment duration or time (0.36 ng/ml versus 0.41 ng/ml; 4 days versus 10 days post-implant, P > 0.1). The pattern of follicle growth and ovulation did not differ significantly between treatment durations and pooled means were used for comparative descriptions. The emergence of a new follicle wave could be detected beginning at 4 days after implant removal (mean 7.9 +/- 0.8 days). After emergence, a period of rapid follicle growth generally ensued with signs of oestrus occurring when the follicle reached 12.3 +/- 0.5 mm and ovulation when mean follicle diameter was 13.1 +/- 0.7 mm at 13.6 +/- 1.5 days after implant removal. Oestrus preceded ovulation in all cases where ovulation subsequently occurred. The mean interovulatory interval after implant removal was similar amongst groups (18.2 +/- 1.3 days). Follicle growth could be categorized into three groups based on the time to emergence and fate of the first wave DF. Spontaneous recovery was characterised by ovulation of the newly emerged DF. Failure to ovulate the first DF was associated with the formation of a persistent follicle by 35 days after implant removal with some 25% (7/28) of cows showing persistent follicles. Delayed emergence (>14 days after implant removal) was detected in 11% (3/28) of cows, but when a follicle did eventually emerge it was seen to ovulate normally. The correlation between number of days to first oestrus expression and LH pulse amplitude on 4 days post-implant removal was significant (R2 = 44%, P < 0.05). A model was then proposed for the restoration of reproductive function following GnRH agonist removal. In conclusion, duration of deslorelin treatment was associated with only small changes in LH and follicle parameters. The formation of persistent follicles delayed the recovery of more cows than delayed emergence of a new follicle wave after implant removal. PMID- 15885438 TI - Comparison of electroejaculation and transrectal massage for semen collection in range and yearling feedlot beef bulls. AB - Two experiments were conducted to compare electroejaculation (EE) and transrectal massage (RM) of the ampullary region for semen collection from beef bulls, and to determine the effect of semen collection method on semen traits. In experiment 1, semen was collected either by EE or RM randomly assigned on an alternate basis in 137 range beef bulls unaccustomed to being handled. The maximum time allowed for RM was 4 min and if no semen was obtained, EE was used. In experiment 2, semen was collected from 39 yearling feedlot beef bulls that were accustomed to being handled, by RM followed immediately by EE. The maximum time allowed for semen collection by both methods was 4 min. In both experiments, sperm concentration, percent of progressively motile sperm, percent of sperm staining alive, and sperm morphology were determined. In experiment 1, RM resulted in fewer (P<0.001) successful semen collections and fewer bulls with penile protrusion than EE (80.9% versus 100% and 54.4% versus 91.5%, respectively). The success of RM was not influenced by bull age or breed, or by the veterinarian performing the massage. Transrectal massage required more time (30s, P<0.001) for obtaining a semen sample and resulted in samples with lower sperm concentration (P<0.001), percent motile sperm (P<0.05) and percent live sperm (P<0.001) when compared to EE. In experiment 2, EE and RM were equally effective for obtaining a semen sample (97.4 and 94.9%, respectively), but the proportion of bulls exhibiting penile protrusion during semen collection was lower (P<0.0001) with RM compared to EE. Percent of sperm staining alive was also lower (P<0.01) in samples collected by RM. Sperm morphology (normal sperm, head defects, midpiece defects, proximal cytoplasmic droplets, and detached sperm heads) did not differ between samples collected by EE and RM. In conclusion, semen could be collected by transrectal massage from approximately 80% of range beef bulls and from 95% of yearling beef bulls accustomed to handling. Sperm morphology was not affected by the method of semen collection, but percent of motile sperm and live sperm were lower in samples collected by RM. A reduced ability to stimulate penile protrusion with RM precluded examination of the penis in a large proportion of bulls. PMID- 15885439 TI - Effect of non-esterified fatty acids on bovine granulosa cell steroidogenesis and proliferation in vitro. AB - In high-yielding dairy cows, the negative energy balance (NEB) during the first weeks post partum may influence dominant follicle growth and steroidogenesis. Since non-esterified fatty acid (NEFA) concentrations are elevated during NEB and are shown to be toxic for several cell types, we investigated the individual and combined effects of the three main NEFA's on granulosa cell proliferation and steroidogenesis in vitro. Granulosa cells from large follicles were cultured for two days in serum free medium in the presence of palmitic (C16:0) (PA), stearic (C18:0) (SA) and/or oleic acid (C18:1) (OA). Addition of 150, 300 or 500 microM of PA and SA inhibited cell proliferation (P<0.05) while OA only elicited such an effect at 500 microM (P<0.01). In the combination treatment (150 microM of each fatty acid), cell numbers were also reduced (P<0.01). These inhibitory effects on cell number are partly due to the induction of apoptosis by these NEFA's, as was demonstrated by annexin V-FITC/propidium iodide staining of the granulosa cells. Oestradiol-17beta production was stimulated by all doses of PA, by 300 and 500 microM of SA and by 500 microM of OA (P<0.05). Combined treatment with 150 microM of each fatty acid also stimulated oestradiol-17beta production per 10(4) cells (P<0.05). We can conclude that PA, SA and to a lesser degree OA modulate granulosa cell proliferation and steroidogenesis in vitro. These effects may be involved in the occurrence of ovarian dysfunction during the postpartum period in high-yielding dairy cows. PMID- 15885440 TI - Morphometric and ultrastructural characterization of Bos indicus preantral follicles. AB - The aim of the present study was to characterize the ultrastructure of zebu cow preantral follicles (PAFs). Ovarian cortex samples were processed for light and transmission electron microscopy. Primordial follicles consisted of an oocyte surrounded by one layer of flattened or flattened-cuboidal granulosa cells. The oocyte contained a large and usually eccentric nucleus. Most organelles were located at the perinuclear ooplasm. Round shaped mitochondria, which contained electron-dense granules, smooth and rough endoplasma reticulum and a Golgi apparatus were also observed. Vesicles and coated pits were often observed in the cortical ooplasm. In primary follicles, the oocyte was surrounded by one layer of cuboidal granulosa cells. Short microvilli were observed on the oolema. Secondary follicles consisted of an oocyte surrounded by a variable number of layers of cuboidal granulosa cells. Small secondary follicles had an ultrastructure very similar to that observed in primary follicles. At this follicular stage, the zona pellucida was beginning to form around the oocyte. In large secondary follicles, the zona pellucida was totally developed around the oocyte. Several granulosa cell projections could be detected that were encroaching into the zona pellucida and protruding towards the oocyte, where gap junctions were observed between oocyte and granulosa cell membranes. Organelles within the oocyte were located at the periphery of the ooplasm, and clusters of cortical granules were observed. Round mitochondria were abundant in all developmental stages. In conclusion, this study described the ultrastructure of zebu cow PAFs, and some unique characteristics could be observed as compared with what has been reported for follicles of Bos taurus cattle. PMID- 15885441 TI - Comparison of three estrus detection systems during summer in a large commercial dairy herd. AB - The objective of the study was to compare three systems for estrus detection and combinations of these systems on a large commercial dairy (1075 lactating cows) during stress of summer heat. At 37-45 days in milk (DIM), 255 cows were fitted with a HeatWatch device (HW; DDx Inc., Denver, CO), an activity sensor ALPRO (ALPRO; DeLaval Inc., Kansas City, MO), and visually observed (VO) three times daily. Pregnancy status was determined by uterine palpation per rectum 35-49 days following artificial insemination (AI). Effects of DIM, parity, standing events, inseminator, and interval between onset of estrus and AI on conception rates were determined using logistic regression. Efficiencies for detection of estrus, determined by comparing detected periods of estrus with a theoretical total of 570 periods, were 49.3% (VO), 37.2% (ALPRO), 48.0% (HW), and 80.2% for all three systems simultaneously. Conception rates (LSM+/-S.E.) for cows detected by one or more of the three systems were 6.2+/-3.9 for VO, 19.8+/-5.6 for ALPRO, 17.3+/-5.0 for HW, 22.8+/-7.0 for VO+ALPRO, 26.9+/-4.6 for VO+HW, 23.2+/-5.2 for ALPRO+HW, and 18.4+/-4.7 for VO+ALPRO+HW. Inseminations performed during no and mild heat stress (temperature-humidity index; THI< or =76) had greater conception rate (P<0.05; 38.8%) compared to AI performed during moderated heat stress conditions (THI>76; 17.6%). Number of mounts were higher for primiparous versus multiparous cows (P<0.05). Cows over 80 DIM during estrus exhibited fewer (P<0.05) standing events. The highest conception rate occurred with the combination of VO+HW, which confirms the premise that combination of multiple systems enhances both the efficiency and accuracy of estrus detection. PMID- 15885442 TI - Infertility in the dromedary bull: a review of causes, relations and implications. AB - Research into infertility in the dromedary bull, as reported during the last two decades, is reviewed with emphasis on causes and effects. Reproductive activity of such animals is naturally limited by a breeding season, though with enough encouragement some may mate with oestrous females out of season but a full fertilization potential can in no way be expected. It is essential that any female presented to a bull is capable of reproducing. The presentation of a subfertile or infertile female due to infection or physiological abnormality will adversely affect the female's ability to conceive and, therefore, the apparent fertility rates of the bull she was put to. The average number of successful services a bull could be expected to perform is two per day. Dromedary bulls with large testes have larger sperm outputs and can cope with more than two females per day providing that they are given adequate periods of rest, 1-2 days every 10 days or so, in conjunction with appropriate nutrition throughout the season. Anabolic steroids or testosterone therapies, which are sometimes used in an attempt to improve male characteristics and bull libido, are not recommended for dromedary bulls in breeding work. Such steroids result in a decrease in testicular size and weight with fewer sperm per gram of testicular tissue being found and the sperm produced also have lower motility rates. Pain associated with the act of mating a she-camel, due to injuries or inflammation in the scrotum, testes, prepuce and sheath, can cause a permanent reduction in bull libido. Camel bulls achieving pregnancy rates more than 60% have had consistently higher spermatozoal concentrations and kinematic variables derived by the computerized cell motion analyzer (CMA) system. As far as physical capabilities are concerned, 3-year-old dromedary bulls, which have reached puberty, have been shown to be perfectly capable of fertilizing a female, but they have a limited sperm production to perform consistently throughout the season in a large herd. By 4.5 5.0 years of age, they are capable of producing adequate numbers of sperm to mate as many as females as an adult bull but fertilizing capacity is not attained until 6 years of age on average. Hyperoestrogenaemia, associated with autoimmune thyroiditis and trypanosomiasis, suppresses the secretion of testicular testosterone and augments the release of testicular histamine, which appears mandatory for quantitative reduction/loss of advanced spermatogenic cells in infertile dromedary bulls. PMID- 15885443 TI - The freemartin syndrome: an update. AB - The freemartin condition represents the most frequent form of intersexuality found in cattle, and occasionally other species. This review considers the current state of knowledge of freemartin biology, incidence, experimental models, diagnosis, uses for freemartins in cattle herds, occurrence in non-bovine species, effects on the male, and highlights potential new research areas. Freemartins arise when vascular connections form between the placentae of developing heterosexual twin foeti, XX/XY chimerism develops, and ultimately there is masculinisation of the female tubular reproductive tract to varying degrees. With twinning rates in Holstein cows increasing, there will be greater economic importance to establish early diagnosis of the freemartin and the detection of the less common single born freemartin. New diagnostic methods based on the detection of Y-chromosome DNA segments by polymerase chain reaction (PCR) show improved assay sensitivity and efficiency over karyotyping and clinical examination. The implications for the chimeric male animal born co-twin to the freemartin are contentious as to whether fertility is affected; if germ cell chimerism does indeed occur; and, if there are any real effects on the sex ratio of offspring produced. In beef cattle, the freemartin carcass has similar characteristics to normal herdmates. Hormonal treatment of freemartins for use as oestrous detectors has been used to obtain salvage value. The biology of freemartin sheep has recently been studied in detail, and the condition may be increasing in prevalence with the introduction of high fecundity genes into flocks. Potential new research areas are discussed, such as detection of foetal DNA in maternal circulation for prenatal diagnosis and investigation of the anti tumour properties of Mullerian inhibiting substance (MIS). The freemartin syndrome will always be a limiting factor in cattle and to a lesser extent in sheep production systems that have the goal to produce multiple reproductively normal female offspring from a single dam without using sex predetermination. PMID- 15885444 TI - Motility and fertility of alginate encapsulated boar spermatozoa. AB - Ejaculated boar spermatozoa are vulnerable to cold shock. Prolonged storage of boar spermatozoa at low temperatures reduces survival rate, resulting in a bottleneck for the extension of artificial insemination in pig husbandry. This study evaluated whether alginate microencapsulization processing can improve the longevity of boar spermatozoa stored at 5 degrees C and the fertility of microencapsulated spermatozoa in vivo. Sperm-rich fraction semen from three purebred boars were concentrated and microencapsulated using alginate at 16-18 degrees C, and then were stored at 5 degrees C. Following storage for 1, 3 and 7 days, the microcapsule was taken out to assess sperm release under 37 degrees C incubation with or without 110 rpm stirring. The percentage of sperm released from microcapsules with 110 rpm stirring was higher than without stirring (81 versus 60%) after 24h of incubation. In another experiment, semen was also microencapsulated to evaluate the sperm motility. The motility of spermatozoa was assessed at 10 min, 8, 24, 32, 48, 56 and 72 h following incubation at 37 degrees C for nine consecutive days. The fertility of the free and microencapsulated semen was assessed by inseminating sows, and the reproductive traits (conception rate, farrowing rate, and litter size) were recorded. The motility of encapsulated spermatozoa was significantly higher than that of free semen after 8h incubation at 37 degrees C after storing for over three days (P<0.05). No significant difference existed in conception rate, farrowing rate, and litter size between the microencapsulated and non-encapsulated semen after four days of storage. In conclusion, microencapsulation can increase the longevity of boar spermatozoa and may sustain in vivo ova fertilization ability. PMID- 15885445 TI - Effect of cooling and seminal plasma on the capacitation status of fresh boar sperm as determined using chlortetracycline assay. AB - Insemination of sows with frozen-thawed spermatozoa results in lower fertility, in part due to spermatozoa having undergone a capacitation-like reaction. The present study employed chlortetracycline (CTC) staining analysis to investigate the effect of adding 20% (v/v) boar seminal plasma (SP) to boar spermatozoa on the temporal progress of capacitation and the acrosome reaction in spermatozoa cooled to 5 degrees C or incubated at 39 degrees C. Based on CTC staining patterns, seminal plasma appeared to reverse capacitation in spermatozoa that had undergone capacitation while incubated at 39 degrees C in a capacitation supporting medium from 59.7 to 36.6% capacitated (P<0.001). Similarly, the addition of SP to boar spermatozoa cooled to 5 degrees C resulted in both the prevention of the capacitation-like reaction, and the reversal of an established capacitation-like reaction from 63.3 to 34.2% capacitated (P<0.001). These observations indicated that some constituent(s) of boar SP both prevent spermatozoa from undergoing capacitation as well as reverse capacitation in spermatozoa that have already undergone the process. PMID- 15885446 TI - High oxygen tension during in vitro oocyte maturation improves in vitro development of porcine oocytes after fertilization. AB - This study was conducted to evaluate the effect of oxygen tension during IVM and/or IVC on developmental competence of porcine follicular oocytes. Prospective, randomized experiments were designed, and oocytes were matured, inseminated and cultured in vitro in the designated condition. In experiment 1, either high (20%) or low (7%) oxygen tension was used for IVM. The high oxygen significantly improved blastocyst formation (23% versus 13%; P<0.01) after IVF than the low oxygen. Such treatment, however, did not significantly (P>0.05) improve the rates of nuclear maturation (89% in each treatment), sperm penetration (62-72%), monospermic fertilization (56-67%), pronuclear formation (90-96%), cleavage (49-53%) and blastocyst cell number (31-32 cells). In experiment 2, the combined effect of oxygen tension during IVM and IVC of embryos was evaluated by a 2 x 2 factorial arrangement. Again, the high oxygen tension during IVM supported blastocyst formation more efficiently (P<0.01) than the low oxygen, and this was independent of oxygen tension during IVC (26-28% versus 15 16%). In oocytes matured under the high oxygen, a tendency to increase blastomere number (P=0.0630) was found, when the low oxygen was used for IVC after insemination (39-45 cells/blastocyst). In conclusion, the use of high oxygen tension (20% maintained by exposure to 5% CO2 in air) for IVM of porcine oocytes promoted blastocyst formation in vitro. PMID- 15885447 TI - Improving post-thaw survival of cryopreserved mouse blastocysts by hydrostatic pressure challenge. AB - The purpose of the experiments was to study the effect of high hydrostatic pressure treatment prior to vitrification to the survival of expanded mouse blastocysts. High hydrostatic pressure has been reported to induce the production of "shock proteins" in bacteria, which can provide a possibility of cross protection to other environmental stresses. The possible beneficial effects of this alleged principle was examined on embryo vitrification. First, the behaviour of blastocysts was studied at altered pressure conditions. In the second part of the study, pressure treatment was combined with a cryopreservation protocol. Our results indicate that the survival of pressurized mouse embryos depends on the magnitude and the duration of pressure applied. We demonstrated that a preceding pressure treatment strikingly increases the survival of the frozen blastocysts as well as the speed of resumption of the development, and hatching rate. PMID- 15885448 TI - 3-Ethyl-2,7-dimethyl octane, a testosterone dependent unique urinary sex pheromone in male mouse (Mus musculus). AB - A previous investigation revealed that urine from normal male mice contained five unique volatile constituents; namely: 3-cyclohexene-1-methanol (I); 3-amino triazole (II); 4-ethyl phenol (III); 3-ethyl-2,7-dimethyl octane (IV); 1 iodoundecane (V). The present study was designed to find out whether the production of these male specific urinary compounds was androgen-dependent. Urine of castrated and castrated plus testosterone-treated male mice was analyzed using gas chromatography linked mass spectrometry (GC-MS). Even though castrated male urine contained 10 detectable compounds, the five male specific compounds present in intact males were absent in castrated male mice urine. Only 3-ethyl-2,7 dimethyl octane (IV) reappeared following testosterone treatment into castrated males. Our earlier bioassay revealed that this compound was involved in attracting females. The present study concluded that this compound was a male specific volatile cue that acted as a releaser pheromone and its production was under the control of androgen. PMID- 15885449 TI - Investigations on the expression of cytokines in the canine corpus luteum in relation to dioestrus. AB - Control of luteal regression in the dog is still poorly understood. Unlike other domestic animal species, luteolysis is not prevented by hysterectomy. Indications that PGF(2alpha) may act as an endogenous luteolytic agent have been found only in pregnant animals during the prepartal decline of progesterone. Evidence from several species indicates that the immune system plays an important role in corpus luteum (Cl) function, possibly by the release of cytokines from immigrant immune cells. Hence, in the present experiment we attempted to examine the expression of cytokines in the canine Cl during the course of dioestrus (formation and regression of the Cl), using RT-PCR. Groups of 4-5 bitches were ovario-hysterectomised on days 5, 15, 25, 35, 45 and 60-80 after ovulation. Canine-specific primers for IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, TNFalpha, IFNgamma and TGFbeta1 were used. Positive and negative controls were included in all tests. Adequate expression was confirmed by sequencing selective samples of positive PCR products. The expression of mRNA for IL-8, IL-10, IL-12, TNF-alpha and TGF-beta1 was detected in all samples at each stage of dioestrus, without any obvious variations indicating a likely modulatory function of these cytokines in differentiation, maintenance or regression of the canine Cl. All tests for the expression of mRNA for IL-4, IL-1beta and IL-2 were negative. More negative than positive results were obtained when testing for the expression of mRNA for IL-6 and IFN-gamma, leading to the conclusion that expression of these two cytokines is at a low level, and no conclusion can be drawn as to their involvement in control of Cl function. PMID- 15885450 TI - Xanthan and galactomannan (from M. scabrella) matrix tablets for oral controlled delivery of theophylline. AB - Directly compressed theophylline tablets, containing commercial xanthan (X) (Keltrol) and a highly hydrophilic galactomannan (G) from the seeds of Mimosa scabrella (a brazilian leguminous tree called bracatinga) as release-controlling agents, were obtained. Gums were used at 4, 8, 12.5 and 25% (w/w), either alone or in mixture (X:G 1:1). During galactomannan extraction process, the biopolymer was dried in a scale up, by vacuum oven (VO) or spray dryer (SD). The in vitro drug release was evaluated at different time intervals during 8 h using apparatus 1 (USP 26) at 100 rpm. The pH of the dissolution medium (1.4) was changed to 4.0 and 6.8 after 2 and 3 h, respectively. Tablets containing G(SD) resulted in more uniform drug release than G(VO) ones, due to their smaller particle size. The drug release decreased with the increase of polymer concentration and all formulations at 25% w/w of gums showed excessive sustained release effect. The matrices made with alone X showed higher drug retention for all concentrations, compared with G matrices that released the drug too fast. The XG matrices were able to produce near zero-order drug release. The XG(SD) 8% tablets provided the required release rate (about 90% at the end of 8 h), with zero-order release kinetics. Tablets containing G(VO) in low concentration showed a complete erosion, while the others demonstrated fast hydration and swelling in contact with the dissolution medium. The release mechanism was a combination of diffusion and relaxation. The relative importance of these two processes varied with matrix composition. The XG(SD) 8% matrix showed higher contribution of polymer relaxation. PMID- 15885451 TI - Aerosolized liposome-based delivery of amphotericin B to alveolar macrophages. AB - The present study was aimed at preparation, characterization, and performance evaluation of amphotericin B (Amp B)-loaded aerosolized liposomes for their selective presentation to lungs (alveolar macrophages), that being the densest site of Aspergillosis infection. Egg phosphatidylcholine (PC)- and cholesterol (Chol)-based liposomes were modified by coating them with alveolar macrophage specific ligands (O-palmitoyl mannan, OPM, and O-polmitoyl pullulan, OPP). The prepared formulations were characterized in vitro for vesicle morphology, mean vesicle size, vesicle size distribution and percent drug entrapment. Pressurized packed systems based on preformed liposomal formulations in chlorofluorocarbon aerosol propellants were prepared. In vitro airways penetration efficiency of the liposomal aerosols was determined by percent dose reaching the peripheral airways, it was recorded 1.4-1.6 times lower as compared to plain drug solution based aerosol. In vivo tissue distribution studies on albino rats suggested the preferential accumulation of OPM- and OPP-coated formulations in the lung macrophages. Higher lung drug concentration was recorded in case of ligand anchored liposomal aerosols as compared to plain drug solution and plain liposome based aerosols. The drug was estimated in the lung in high concentration even after 24 h. The drug-localization index calculated after 6 h was nearly 1.42-, 4.47-, and 4.16-fold, respectively, for plain, OPM-, and OPP-coated liposomal aerosols as compared to plain drug solution-based aerosols. These results suggest that the ligand anchored liposomal aerosols are not only effective in rapid attainment of high-drug concentration in lungs with high population of alveolar macrophages but also maintain the same over prolonged period of time. The significance of targeting potential of the developed systems was established. PMID- 15885452 TI - The influence of dose on the performance of dry powder inhalation systems. AB - The relationship between drug/lactose ratio and aerosolisation performance of conventional carrier based formulations was investigated using the twin stage impinger. A dose range of approximately 10-450 microg of drug in a 50 mg lactose carrier formulation was studied. Statistical differences in both the fine particle dose and fine particle fraction were observed across the dosage range (ANOVA, p<0.05). In general, no statistically significant difference (Fishers Pairwise, p<0.05) in fine particle dose was observed between drug levels of approximately 10 microg and 135 microg, whereas a linear decrease in fine particle fraction was observed across the same drug level range (R2=0.977). Increasing the dose from approximately 135 microg to 450 microg resulted in a statistically significant increase in both fine particle dose and fraction (ANOVA p<0.05). Such observations may be attributed to the occupation of 'active' carrier sites by drug particles at low drug concentration, since the quantity of drug particles liberated from the carrier during aerosolisation remains constant at the lower dosing regimes. PMID- 15885453 TI - Scintigraphic imaging of radiolabelled drug delivery systems in rabbits with arthritis. AB - This paper describes a novel approach for designing drug delivery systems for intra-articular (i.a.) treatment of rheumatoid arthritis. Retention of these systems was evaluated by radiolabeling with Tc-99m and gamma scintigraphy in arthritic rabbits. Liposome, niosome, lipogelosome and niogelosome formulations of Diclofenac Sodium (DFNa) have been prepared and drug release properties and in vitro characterisation studies have been carried out. According to characterisation results L1 (DMPC: CHOL: DCP (7:1:2)), L1J1 (DMPC: CHOL: DCP (7:1:2) in C-940 1:1 (w/w)), N (SUR I: CHOL: DCP (7:1:2)) and NJ1 (SUR I: CHOL: DCP (7:1:2) in C-940 1:1 (w/w)) formulations were chosen for the further studies. Retention time of these formulations was evaluated by gamma scintigraphic imaging studies. Rabbits with antigen-induced arthritis were injected intra-articularly with Tc-99m labelled drug delivery systems. Serial scintigraphic images were obtained to investigate the retentions of labelled drug delivery systems in the arthritic joints and choose a suitable formulation for the treatment protocol of arthritis. At the end of the scintigraphic imaging studies it was observed that radiolabelled lipogelosome formulation containing DFNa (L1J1) retained much longer in the experimentally arthritic knee joints of the rabbits. This formulation was used for the treatment protocol of arthritis. Mono articular arthritis was induced in the knee joints of rabbits and it was monitored at regular time intervals by measuring changes in knee joint diameter. Also macroscopic and histopathologic evaluations were performed for further evaluation of arthritis. Great retention of DFNa in the arthritic joint might reduce potential adverse systemic effects of the drug because of local administration into the diseased area. It appeared to be a promising drug delivery system for intra-articular drug delivery. PMID- 15885454 TI - In vivo comparison of various liposome formulations for cosmetic application. AB - The interaction of liposome formulations, prepared with phospholipids of different origins (egg and soya), with skin were compared in terms of their effects on skin water content, skin barrier function, and skin elasticity. Short term effect of four different liposome formulations and two references during 3.5 h was investigated non-occlusively on the volar side of the forearm of 10 volunteers, ranging in age from 24 to 32 years. Liposomes composed of different phospholipids showed differing effects on skin humidity. The maximal effect was achieved within 30 min and constant values were reached after 1.5 h for all formulations, however values remained significantly higher than without treatment (p<0.05) during the whole application time. The best results were obtained with liposome formulations prepared from egg phospholipids, which exhibited a 1.5-fold increase in skin water content (p<0.05), whereas liposome formulations prepared from soya phospholipids showed no advantage compared to the references. Skin barrier function showed greatest influence within 30 min after application and remained constant after 1.5 h for all formulations. Within the liposome formulations, egg phospholipids showed the highest transepidermal water loss values during the first 30 min, representing the strongest interactions with the skin barrier function, whereas for the other liposome formulations lower transepidermal water loss values were measured. Skin elasticity and tiring effect of the skin was not influenced by any of the formulations, due to the young skin tested. Long-term effect of two different liposome formulations mixed with base cream DAC in two different concentrations during 14 days was investigated non occlusively on the volar side of the forearm of 10 volunteers, ranging in age from 20 to 25 years. Skin water content was measured daily and the results showed that skin humidity was increased significantly (p<0.05) for the formulation containing 20% egg phospholipids during 6 days. Liposome formulations prepared with egg phospholipids showed significantly higher (p<0.01) hydration effects during 3.5 h of application on human skin compared with liposome formulations prepared with soya phospholipids. PMID- 15885455 TI - Determination by NMR of the binding constant for the molecular complex between alprostadil and alpha-cyclodextrin. Implications for a freeze-dried formulation. AB - A binding constant was determined for the complexation reaction between alprostadil (PGE1) and alpha-cyclodextrin (alpha-CD). This constant was used to calculate the fraction PGE1 free upon reconstitution of Caverject dual chamber syringe, indicated for the treatment of erectile dysfunction. The determination was based on the measurement of the chemical shift of the C20 methyl protons of PGE1. The observed chemical shift varies as a linear function of the amount of PGE1 bound. The binding constant was obtained from the binding isotherm, a curve of the observed chemical shift versus free ligand (alpha-CD) concentration, through the application of non-linear regression analysis. A value K11 = 966 M( 1) +/- 130 M(-1) (2s), measured at 27 degrees C, was obtained. This value is in good agreement with those reported in the literature. The percent PGE1 free was subsequently calculated for the reconstituted solution and in the corpora cavernosum after injection. The latter showed PGE1 to be delivered essentially quantitatively to the targeted site. PMID- 15885456 TI - Factors affecting the characteristics of rapidly disintegrating tablets in the mouth prepared by the crystalline transition of amorphous sucrose. AB - The aim of this study is to investigate the factors affecting the characteristics of rapidly disintegrating tablets containing an amorphous ingredient prepared by crystalline transition method (CTM) under various storage conditions. Effect of storage conditions and formulating ratio of amorphous sucrose on the characteristic changes (tensile strength, porosity, and disintegration time) of the rapidly disintegrating tablets was studied. The storage conditions of different temperature and humidity affected the rate of crystalline transition and the increase in the tablet tensile strength. The faster crystalline transition resulted in a faster rate of increase in the tablet tensile strength. Regarding the effect of the formulating ratio of amorphous sucrose, in the case of 20-100%, the tensile strength after storage as a function of the porosity could be plotted on the same curve. For tablets containing 100% amorphous sucrose, the tablets with different porosity changed to almost the same structure due to the crystalline transition. Hence, the higher formulating ratio of amorphous sucrose provided the longer disintegration time in the mouth. Therefore, we concluded that the formulating ratio of 10-20% of the amorphous sucrose in the tablet is suitable for the rapidly disintegrating tablet in the mouth when prepared by CTM. PMID- 15885457 TI - In vitro release of diclofenac diethylamine from caprylocaproyl macrogolglycerides based microemulsions. AB - The purpose of the present study was to determine the influence of both formulation parameters and vehicle structure on in vitro release rate of amphiphilic drug diclofenac diethylamine (DDA) from microemulsion vehicles containing PEG-8 caprylic/capric glycerides (surfactant), polyglyceryl-6 dioleate (cosurfactant), isopropyl myristate and water. From the constructed pseudo ternary phase diagram at surfactant-cosurfactant mass ratio (K(m) 1:1), the optimum oil-to-surfactant-cosurfactant mass ratio values (O/SC 0.67-1.64) for formulation of microemulsions with similar concentrations of hydrophilic, lipophilic and amphiphilic phases (balanced microemulsions) were found. The results of characterization experiments indicated bicontinuous or nonspherical water-continuous internal structure of the selected microemulsion vehicles. Low water/isopropyl myristate apparent partition coefficient for DDA as well as elevated electrical conductivity and apparent viscosity values for the investigated microemulsion formulations containing 1.16% (w/w) of DDA, suggested that the drug molecules was predominantly partitioned in the water phase and most likely selfaggregate and interact with interfacial film. Release of DDA from the selected water-continuous (W/O), oil-continuous (O/W) and balanced microemulsions was investigated using rotating paddle dissolution apparatus modified by addition of enhancer cell. A linear diffusion of DDA through regenerated cellulose membrane was observed for the W/O and O/W formulations with the low content of dispersed phase. Non-linearity of the drug release profile in the case of bicontinuous formulations was related to the more complex distribution of DDA including interactions between the drug and vehicle. The membrane flux value increases from 25.02 microgcm(-2)h(-1) (W/O microemulsion) to 117.94 microgcm( 2)h(-1) (O/W microemulsion) as the water phase concentration increases. Moreover, the obtained flux values for balanced microemulsions (29.38-63.70 microgcm(-2)h( 1)) suggested that bicontinuous microstructure hampers the release of the amphiphilic drug. PMID- 15885458 TI - Non-ionic surfactant based vesicles (niosomes) for non-invasive topical genetic immunization against hepatitis B. AB - DNA vaccines are capable of eliciting both humoral as well as cellular immune responses. Liposomes have been widely employed for DNA delivery through topical route; however, they suffer from certain drawbacks like higher cost and instability. In present study, non-ionic surfactant based vesicles (niosomes) for topical DNA delivery have been developed. DNA encoding hepatitis B surface antigen (HBsAg) was encapsulated in niosomes. Niosomes composed of span 85 and cholesterol as constitutive lipids were prepared by reverse phase evaporation method. Prepared niosomes were characterized for their size, shape and entrapment efficiency. The immune stimulating activity was studied by measuring serum anti HBsAg titer and cyokines level (IL-2 and IFN-gamma) following topical application of niosomes in Balb/c mice and results were compared with naked DNA and liposomes encapsulated DNA applied topically as well as naked DNA and pure recombinant HBsAg administered intramuscularly. It was observed that topical niosomes elicited a comparable serum antibody titer and endogenous cytokines levels as compared to intramuscular recombinant HBsAg and topical liposomes. The study signifies the potential of niosomes as DNA vaccine carriers for effective topical immunization. The proposed system is simple, stable and cost effective compared to liposomes. PMID- 15885459 TI - The delayed dissolution of paracetamol products in the canine fed stomach can be predicted in vitro but it does not affect the onset of plasma levels. AB - Although it is generally believed that paracetamol can be used as a marker of gastric emptying, there have been reports in the literature that show delayed dissolution of immediate release paracetamol tablets using standard in vitro setups and food-simulating media, delayed disintegration of paracetamol products in the fed stomach, and no correlation of paracetamol absorption with gastric emptying in the fed state. In this study, we confirmed that dissolution of Panodil and Apotel tablets is delayed in food-simulating media regardless of the in vitro hydrodynamics and on a formulation dependent manner. Further, we assessed the usefulness of in vitro dissolution data in the prediction of delayed disintegration time in the fed stomach and we examined the importance of delayed gastric disintegration on the onset of plasma levels using the canine model. In vitro dissolution data in cow's milk reflected the delayed disintegration of Panodil tablets in the fed stomach. In vitro dissolution of Apotel tablets in milk was delayed less than of Panodil and the effect of dosing conditions on the in vivo disintegration was not apparent. However, for the products tested in this study, there was no correlation between intragastric disintegration and onset of plasma levels probably because gastric emptying in also delayed in the fed state. PMID- 15885460 TI - Pharmacokinetics of DA-8159, a new erectogenic, administered at 10:00 h versus 22:00 h in rats. AB - Little is known about chronopharmacokinetics of PDE V inhibitors in rats as well as in humans. Hence, the pharmacokinetics of DA-8159 and one of its metabolites, DA-8164, were investigated after intravenous and oral administration of DA-8159 at a dose of 30 mg/kg administered at 10:00 h versus 22:00 h in rats. After intravenous administration of DA-8159 at 22:00 h, the AUC of DA-8159 was significantly greater (528 versus 368 microg min/ml) due to significantly slower CL (56.1 versus 79.5 ml/min/kg) in the rats. After intravenous administration of DA-8159 at 22:00 h, the AUC of DA-8164 was also significantly greater (108 versus 66.8 microg min/ml) possibly due to significantly greater exposure of the parent drug (AUC of DA-8159). After intravenous administration of DA-8164 at 22:00 h, the CL of DA-8164 was significantly slower; hence, this factor could also contribute to the greater AUC of DA-8164 after intravenous administration of DA 8159. However, after oral administration of DA-8159, the AUC values of both DA 8159 and DA-8164 were not significantly different between 10:00 h and 22:00 h. This was not due to decrease in gastrointestinal absorption of DA-8159 at 22:00 h and may be due to changes in intestinal first-pass effect at 22:00 h. The above data suggested that modification of dosage regimen of oral DA-8159 is not necessary in humans between 10:00 h and 22:00 h. Further studies are needed in humans. PMID- 15885461 TI - Development of a mucoadhesive and permeation enhancing buccal delivery system for PACAP (pituitary adenylate cyclase-activating polypeptide). AB - The buccal mucosa providing direct entry into the systemic circulation appears to be a potential site for the delivery of PACAP (pituitary adenylate cyclase activating polypeptide), a new therapeutic agent in the treatment of type 2 diabetes. In order to reach a sufficient buccal bioavailability a drug delivery system with strong permeation enhancing and mucoadhesive properties is needed. In this study the enhancing effect of a strongly mucoadhesive chitosan-thioglycolic acid (TGA) conjugate in combination with reduced glutathione (GSH) on the permeation of PACAP across the buccal mucosa was investigated. The apparent permeability coefficient (P(app)) of PACAP in buffer only was (5.7 +/- 3.1) x 10( 8), while in the presence of chitosan-TGA conjugate (1%) a P(app) of (20.0 +/- 3.4) x 10(-8) was achieved. The combination of chitosan-TGA (1%) with GSH (2%) led to an improvement of the P(app) up to (57.3 +/- 31.7) x 10(-8). Release studies of PACAP demonstrated that a controlled release can be provided from tablets consisting of chitosan-TGA at a pH of 5, whereas more than twice as much was released from chitosan-TGA tablets pH 4. According to the combination of permeation enhancing properties, controlled drug release and the mucoadhesive character, chitosan-TGA conjugates represent a promising tool for the buccal administration of PACAP. PMID- 15885462 TI - Surface morphology control of polylactide microspheres enclosing irinotecan hydrochloride. AB - In order to reduce the initial burst from polylactide (PLA) microspheres enclosing an antitumor agent, we prepared the microspheres with a smooth surface by varying solvent evaporation conditions such as operating temperature and pressure. PLA microspheres enclosing irinotecan hydrochloride (CPT) were prepared using the O/O emulsion system for solvent evaporation. The mean diameter and enclosing efficiency were almost constant because they were independent of solvent evaporation conditions. Scanning electron microscopic (SEM) observation verified the smooth surface of the PLA microspheres produced by varying the preparation conditions. In vitro release experiments show that the initial burst of microspheres with a smooth surface was less than that of those with a rough surface. PMID- 15885463 TI - Immobilisation of vaccines onto micro-crystals for enhanced thermal stability. AB - The thermal instability of many vaccines leads to the wastage of half of all supplied vaccines. In this note, we report the application of a novel technology: protein-coated micro-crystals (PCMC) to improve the thermostability of a model vaccine (diphtheria toxoid, DT). The latter was immobilised onto the surface of a crystalline material (L-glutamine) via a rapid dehydration method, resulting in the production of a fine free-flowing powder. The PCMC consisted of thin, flat crystals with an antigen loading of 3.95% (w/w). The DT-coated glutamine crystals and free DT (the controls) were incubated at different temperatures for a defined time period (4 degrees C, RT and 37 degrees C for 2 weeks and 45 degrees C for 2 days), after which the crystals were suspended in buffer and intramuscularly administered to mice. Incubation of DT (free and crystal-coated) at room temperature and at 37 degrees C for 2 weeks did not result in any change in the antibody response compared to DT that had always been stored properly (i.e. in the refrigerator). In contrast, incubation of free DT at 45 degrees C resulted in a reduced IgG response, indicating thermal instability of free DT at that temperature. The antibody response was not reduced, however, with the crystal coated DT. These preliminary studies show that PCMC is a promising technology for the thermal stabilisation of vaccines. PMID- 15885464 TI - New surface-modified lipid nanoparticles as delivery vehicles for salmon calcitonin. AB - The aim of the present work was to develop a new nanoparticulate system, consisting of lipid nanoparticles coated with chitosan (CS), intended for the oral administration of peptide drugs. These new nanoparticles were studied, and compared with the previously developed PEG-coated lipid nanoparticles, with regard to their ability to incorporate and deliver the model peptide salmon calcitonin (sCT). Moreover, the influence of the core composition, either a solid triglyceride (tripalmitin), or a mixture of a liquid and a solid triglyceride (Miglyol 812 and tripalmitin) on the encapsulation and release of sCT was investigated. The results showed that a CS coating could be formed around the tripalmitin nanoparticles by simple incubation of the lipid cores in a CS solution, due to the high affinity of CS for the lipid core. In addition, sCT could be efficiently associated, irrespective of the core composition, to the nanoparticles. This important association was attributed to the marked affinity of sCT for the lipid cores, as confirmed by the adsorption studies. However, the nature of the coating affected the surface association of the peptide, which was less important for the nanoparticles coated with CS, than for PEG-coated nanoparticles. This was attributed to the displacement of the sCT molecules located on the surface of the nanoparticles by the positively charged CS molecules. This reduced surface association led to a decrease in the burst release effect, which was more pronounced for the nanoparticles coated with PEG than for those coated with CS. Following the initial burst, the systems provided a continuous and slow release of the associated peptide, independently of the nature of the coating. This slow release was attributed to the affinity of the peptide for the lipids and to the absence of degradation of the lipid matrix under the in vitro release conditions. PMID- 15885465 TI - Applicability of anti-neovascular therapy to drug-resistant tumor: suppression of drug-resistant P388 tumor growth with neovessel-targeted liposomal adriamycin. AB - Anti-neovascular therapy, one of the effective anti-angiogenic chemotherapy, damages new blood vessels by cytotoxic agents delivered to angiogenic endothelial cells and results in indirect eradication of tumor cells. We previously reported that liposomes-modified with a pentapeptide, Ala-Pro-Arg-Pro-Gly (APRPG-Lip) homing to angiogenic site, highly accumulated in tumor tissue, and APRPG-Lip encapsulating adriamycin (APRPG-LipADM) effectively suppressed tumor growth in tumor-bearing mice. In the present study, we examined the topological distribution of fluorescence-labeled APRPG-LipADM as well as TUNEL-stained cells in an actual tumor specimen obtained from Colon 26 NL-17 carcinoma-bearing mice. The fluorescence-labeled APRPG-Lip dominantly localized to vessel-like structure: a part of which was also stained with anti-CD31 antibody. Furthermore, TUNEL stained cells were co-localized to the same structure. These data indicated that APRPG-LipADM bound to angiogenic endothelial cells and induced apoptosis of them. We also investigated the applicability of anti-neovascular therapy using APRPG LipADM to ADM-resistant P388 solid tumor. As a result, APRPG-LipADM significantly suppressed tumor growth in mice bearing the ADM-resistant tumor. These data suggest that APRPG-LipADM is applicable to various kinds of tumor including drug resistant tumor since it targets angiogenic endothelial cells instead of tumor cells, and eradicates tumor cells through damaging the neovessels. PMID- 15885466 TI - Construction of a multifunctional envelope-type nano device by a SUV*-fusion method. AB - A novel assembly method "SUV*-fusion method" was developed for the construction of a small and homogenous multifunctional envelope-type nano device (MEND) by utilizing a detergent-rich small unilamellar vesicle (SUV*). The method consists of three steps: (1) DNA condensation with a polycation, (2) electrostatic interaction of the SUV* with the DNA/polycation complex (DPC) and (3) lipid coating of DPC by SUV* fusion via removal of the detergent. We confirmed the construction of the MEND by sucrose density gradient centrifugation, and isolated the MEND only from the boundary between 25% and 40% sucrose. The isolated MEND had a small diameter (155 nm), was negatively charged (-24 mV), and encapsulated 30% of the total DNA. The MEND was formed by only SUV*, not by a lipid/detergent micelle. This confirms that a small and homogenous MEND can be constructed by the SUV*-fusion method. Furthermore, we confirmed that a transferrin-modified MEND could deliver a gene into a cell through receptor-mediated endocytosis. Consequently, we report on the successful construction of a small and homogenous MEND by a novel SUV*-fusion method. PMID- 15885467 TI - Cellular recognition of paclitaxel-loaded polymeric nanoparticles composed of poly(gamma-benzyl L-glutamate) and poly(ethylene glycol) diblock copolymer endcapped with galactose moiety. AB - Poly(gamma-benzyl L-glutamate) (PBLG)/poly(ethylene glycol) (PEG) diblock copolymer endcapped with galactose moiety (abbreviated as GEG) was synthesized and characterized for study of liver-specific targeting. From dynamic light scattering measurement, particle sizes of copolymeric nanoparticles were decreased with an increase of PEG in the copolymer. The morphology of GEG-3 nanoparticles observed by transmission electron micrograph was observed as almost spherical shapes and ranged about 50-300 nm. From the structural characterization using 1H nuclear magnetic resonance, both characteristic peaks of PBLG and PEG were visible in CDCl3 but the characteristic peaks of PBLG were invisible in D2O, indicating that GEG block copolymers are found to the core-shell type nanoparticles in water with PBLG innercore and PEG outershell, exposing that galactose moiety of GEG block copolymers are outerwards oriented on the nanoparticle surfaces. By galactose-specific aggregation test of particles using beta-galactose specific lectin, and flow cytometry measurement, specific interaction between asialoglycoprotein receptors (ASGPR) of HepG2, human hepatoma cell line, and galactose moieties of the GEG nanoparticles was confirmed. From cell cytotoxicity test, HepG2 cells with ASGPR are more sensitive to paclitaxel (TX)-loaded nanoparticles than free TX whereas, P388 cells, murine leukemia cell line, and SK-Hep 01, human hepatoma cell line, without ASGPR is less sensitive to TX-loaded nanoparticles than free TX, suggesting that specific interaction between HepG2 cells and galactose moiety of the nanoparticles occurred. PMID- 15885468 TI - Interleukin-1 modulates periprosthetic tissue formation in an intramedullary model of particle-induced inflammation. AB - Interleukin-1 (IL-1) is a proinflammatory cytokine that has been implicated in wear-debris associated total joint replacement failure. We hypothesized that the absence of the IL-1 type-1 receptor would mitigate the inflammatory response to titanium particles and decrease periprosthetic inflammatory tissue in a murine intramedullary rod model. METHODS: An intramedullary rod with and without commercially pure titanium particles was placed in the femora of 24 wild type mice (WT) and 24 mice lacking a functional type-1 receptor to IL-1. Femora were analyzed histologically and by ELISA of organ culture explant supernatants. RESULTS: The presence of titanium particles in WT mice stimulated increased expression of interleukin-6 (IL-6) and macrophage chemoattractant protein-1 (MCP 1) relative to rod only controls. In contrast, IL-6 and MCP-1 expression were diminished in IL-1r1-KO mice exposed to titanium particles. Additionally, the formation of a periprosthetic fibro-inflammatory membrane in IL-1r1-KO mice was blunted at 2 weeks when compared to that in wild-type mice. Inflammatory changes and the quality of periprosthetic bone of IL-1r1-KO mice was similar to WT mice in response to titanium particles. CONCLUSIONS: These results implicate IL-1 as an important modulator in the local inflammatory response to intramedullary titanium particles. MCP-1 appears to be significantly modulated in IL-1r1-KO mice in response to titanium particles. This may be responsible, in part, for the diminished periprosthetic membrane observed in IL-1r1-KO mice at 2 weeks. Expansion of this murine model of intramedullary particle-induced inflammation to other gene targets may contribute to a more mechanistic understanding of wear debris associated prosthesis failure. PMID- 15885469 TI - Arthroplasty membrane-derived fibroblasts directly induce osteoclast formation and osteolysis in aseptic loosening. AB - PURPOSE: Both macrophages and fibroblasts are the main cell types found in periprosthetic tissues surrounding failed joint arthroplasties. These fibroblasts are known to express RANKL and to produce TNFalpha, factors which promote osteoclast formation and bone resorption. In this study we have analysed the role that arthroplasty membrane-derived fibroblasts (AFb) play in inducing the generation of bone resorbing osteoclasts. METHODS: Fibroblasts were isolated from periprosthetic tissues and co-cultured with human monocytes in an osteoclast differentiation assay in the presence or absence of M-CSF and inhibitors of RANKL (OPG) and/or TNFalpha. RANKL expression by AFbs was determined by RT-PCR and the extent of osteoclast differentiation by the expression of TRAP, VNR and evidence of lacunar resorption. RESULTS: In the presence of M-CSF, large numbers of TRAP(+) and VNR(+) multinucleated cells capable of lacunar resorption, were noted in co-cultures of monocytes and RANKL-expressing AFbs. Cell-cell contact was required for osteoclast formation. The addition of OPG and anti-TNFalpha alone significantly reduced but did not abolish the extent of osteoclast formation, whereas the addition of both together abolished osteoclast formation and lacunar resorption. CONCLUSION: Our results indicate that fibroblasts in periprosthetic tissues are capable of inducing the differentiation of normal human peripheral blood mononuclear cells to mature osteoclasts by a mechanism that involves both RANKL and TNFalpha. Suppression of both RANKL and inflammatory cytokines is likely to be required to control periprosthetic osteolysis. PMID- 15885470 TI - Polyethylene wear rate and osteolysis: critical threshold versus continuous dose response relationship. AB - We studied the relationship between polyethylene wear and osteolysis in 230 subjects after cemented Charnley total hip arthroplasty in order to examine the validity of the wear rate threshold concept. Polyethylene wear measured using image analysis (EBRA) software was compared in 115 subjects with osteolysis versus 115 control subjects that were individually matched for age, sex, and follow up period. Subjects with osteolysis had almost twice the mean annual wear rate versus the controls. The incidence of osteolysis increased in a linear manner with each quintile increase in wear rate throughout the range 0.01-0.54 mm/year. The odds-ratio for osteolysis approximately doubled with each quintile increase in wear rate above the middle quintile (wear rate 0.08-0.11 mm/year), and decreased at a similar rate with each quintile decrease in wear rate below the middle quintile. Our data suggests that the association of osteolysis with polyethylene wear rate represents a continuous dose-response relationship and does not support the concept of a discrete critical wear rate threshold above which the risk of osteolysis is disproportionately increased. PMID- 15885471 TI - Serum levels of cobalt and chromium in patients with Sikomet metal-metal total hip replacements. AB - Metal/metal total hip replacements (M/M THRs) made of CoCr-based alloys were reintroduced in hip arthroplasty in order to avoid the problem of polyethylene wear and consequently to diminish the wear debris-related osteolysis. Emerging concerns accompanying the reintroduction of M/M THRs are related to the possibility increase of Co and Cr serum levels and, therefore, a careful surveillance of M/M patients is recommended. In the present work three groups of patients with Sikomet SM21 low-carbon alloy were investigated: retrospective group, prospective group and revision group. The investigation of these different groups of patients provided data concerning the metal concentration as a function of time in situ, as well as time ex situ, i.e., following the removal of M/M bearing. A retrospective group of 25 patients sampled at average 60 +/- 20 months postoperatively showed a 3-fold increase in cobalt and a 4.4-fold increase in chromium compared with the control group. In the prospective group of seven patients both cobalt and chromium increase with time and at average 140 +/- 80 days achieved a significant increase compared with preoperative values. An important contribution of this study to the current knowledge on M/M bearings is achieved by the measurements in the revision group of 10 patients, where we observed a decline of serum Co and Cr levels following the removal of M/M articulation. While the link between the increase in metal serum levels and the presence of M/M THR is proved, the biological impact of these results remains to be elucidated. PMID- 15885472 TI - Anterior tibial post impingement in a posterior stabilized total knee arthroplasty. AB - Despite the numerous long-term success reports of posterior stabilized (PS) total knee arthroplasty (TKA), recent retrieval studies of various PS TKA designs revealed wear and deformation on the anterior side of the tibial post. This study investigated the mechanisms of anterior impingement of the post with the femoral component. Seven cadaveric knees were tested to study kinematics and tibial post biomechanics during simulated heel strike using an in vitro robotic testing system. Intact knee kinematics and in situ anterior cruciate ligament (ACL) forces were determined at hyperextension (0 degree to -9 degrees) and low flexion angles (0 degrees to 30 degrees) under the applied loads. The same knee was reconstructed using a PS TKA. The kinematics and the tibial post contact forces of the TKA were measured under the same loading condition. The ACL in the intact knee carried load and contributed to knee stability at low flexion angles and hyperextension. After TKA, substantial in situ contact forces (252.4 +/- 173 N at 9 degrees of hyperextension) occurred in the tibial post, indicating anterior impingement with the femoral component. Consequently, the TKA showed less posterior femoral translation compared to the intact knee after the impingement. At 9 degrees of hyperextension, the medial condyle of the intact knee translated 0.1 +/- 1.1 mm whereas the medial condyle of the TKA knee translated 5.6 +/- 6.9 mm anteriorly. The lateral condyle of the intact knee translated 1.5 +/- 1.0 mm anteriorly whereas the lateral condyle of the TKA knee translated 2.1 +/- 5.8 mm anteriorly. The data demonstrated that anterior tibial post impingement functions as a substitute for the ACL during hyperextension, contributing to anterior stability. However, anterior post impingement may result in additional polyethylene wear and tibial post failure. Transmitted impingement forces might cause backside wear and component loosening. Understanding the advantages and disadvantages of the tibial post function at low flexion angles may help to further improve component design and surgical techniques and thus enhance knee stability and component longevity after TKA. PMID- 15885473 TI - The role of hereditary hemochromatosis in aseptic loosening following primary total hip arthroplasty. AB - Hereditary hemochromatosis (HH) results in increased iron absorption and subsequent deposition in tissue. This condition occurs predominantly in individuals of Northern European and Celtic origin with Ireland having one of the highest allele frequencies in the world. This study examines the hypothesis that homozygosity for either the C282Y or H63D mutations in the HFE gene may be associated with aseptic loosening following total hip arthroplasty (THA). Two groups of individuals were screened for the C282Y and H63D mutations associated with HH. Group 1 were individuals who had undergone primary hip arthroplasty and group 2 were individuals who had undergone revision hip arthroplasty for aseptic loosening. Exclusion criteria included rheumatoid or other inflammatory arthropathies and revision due to causes other than aseptic loosening. Significantly more patients in the revision THA group were homozygous for the C282Y genotype (P = 0.014). Aseptic loosening occurred earlier in these patients (P = 0.009), in particular in the patients who had clinical signs of hemochromatosis. No association was seen with the H63D mutation and revision THA. The incidence of HH in the group of primary THA patients was no higher than the background incidence. Patients who require primary THA and who are homozygous for the C282Y mutation have an increased risk of developing aseptic loosening, leading to revision THA. Moreover C282Y homozygosity appears to be associated with earlier aseptic loosening than in individuals without the C282Y mutation. PMID- 15885474 TI - Three-dimensional finite element analysis of unicompartmental knee arthroplasty- the influence of tibial component inclination. AB - Aseptic loosening and failure of a tibial component are recognized problems in unicompartmental knee arthroplasty (UKA). Excessive stress on the supporting cancellous bone is thought to contribute to the loosening and failure. Of factors that could influence supporting cancellous bone stresses, we focused on the inclination of a unicompartmental tibial component by analyzing the effect of coronal plane and sagittal plane inclination. Detailed geometrically accurate, three-dimensional finite element models were constructed from computed tomography (CT) data of a typical adult male proximal tibia. The material properties for the models were obtained directly from the CT data to simulate the inhomogeneous distribution of cancellous bone properties. Placing the component in slight valgus inclination in the coronal plane reduced the cancellous bone stresses. Posterior inclination in the sagittal plane caused a moderate increase in the stresses. Our results suggest that slight valgus inclination of a UKA tibial component may be preferable to varus or square inclination in the coronal plane. An excessive posterior slope of a tibial component should be avoided. PMID- 15885475 TI - Meniscal material properties are minimally affected by matrix stabilization using glutaraldehyde and glycation with ribose. AB - Knee meniscus replacement holds promise, but current allografts are susceptible to biodegradation. Matrix stabilization with glutaraldehyde, a crosslinking agent used clinically to fabricate cardiovascular bioprostheses, or with glycation, a process of crosslinking collagen with sugars such as ribose, is a potential means of rendering tissue resistant to such degradation. However, stabilization should not significantly alter meniscal material properties, which could disturb normal function in the knee. Our objective was to evaluate the effects of glutaraldehyde and glycation-induced matrix stabilization on the material properties of porcine meniscus. Normal untreated meniscus specimens were tested in confined compression at one of three applied stresses (0.069, 0.208, 0.347 MPa), subjected to either a glutaraldehyde or glycation stabilization treatment, and then re-tested to measure changes in tissue aggregate modulus, permeability, and compressive strain at equilibrium. Changes in these properties significantly increased with glutaraldehyde concentration and exposure time to ribose. One glutaraldehyde and three glycation treatments did not alter aggregate modulus or compressive strain at equilibrium compared to controls (p > 0.10). However, all treatments increased permeability by at least 108% compared to controls (p < 0.001). This study reveals a dose-dependent relationship between meniscal material properties and certain stabilization conditions and identifies treatments that minimally affect these properties. Further research is necessary to determine whether these treatments prevent enzymatic degradation before and after surgical implantation in the knee. PMID- 15885476 TI - Proteoglycan 4 (PRG4) synthesis and immunolocalization in bovine meniscus. AB - Proteoglycan 4 (PRG4) is synthesized and secreted into the synovial fluid by articular chondrocytes and synovial cells, lining the cavity of joints. A thin layer of PRG4 is also present at the articular surface, where it appears to be involved in boundary lubrication. This study investigated if PRG4 is also synthesized and secreted by the cells within meniscus, and if PRG4 is also present in, and at the surface of, meniscus. PRG4 was visualized in sections of bovine calf menisci by immunohistochemistry. PRG4 was detected in two regions: (1) at the femoral and tibial surfaces of the meniscus, and within cells below these surfaces; and (2) within and near cells along the radial tie fibers and circumferential fibers. From meniscus tissue harvested from these surfaces, PRG4 was extracted with 4M GuHCl and quantified by ELISA. There was 0.20 +/- 0.01 and 0.25 +/- 0.04 microg PRG4/cm(2) area of lateral and medial meniscus surface, respectively. ELISA analysis of spent medium from other samples of meniscus surface tissue incubated in medium supplemented with serum and ascorbate showed that 8.1 +/- 1.1 microg PRG4/cm(2) area of meniscus surface was secreted over six days. These results demonstrate that PRG4 is synthesized and secreted by certain cell populations in the meniscus, and that PRG4 is present in the meniscus at surfaces and also internal fibers where it may contribute to boundary lubrication. PMID- 15885477 TI - Increased concentrations of neuro-excitatory amino acids in rat anterior cruciate ligament-transected knee joint dialysates: a microdialysis study. AB - Changes in excitatory amino acid (EAA) levels were examined in the knee joint dialysates of rats with early osteoarthritis (OA). Early OA was induced by anterior cruciate ligament (ACL) transection in one knee and the contralateral knee was used as the sham-operated control, the side for ACL transection being assigned randomly. Twenty weeks after operation, knee joint dialysates were collected by microdialysis and assayed for EAAs by high performance liquid chromatography. The rats were then sacrificed for histopathological examination. Hematoxylin/eosin and Safranin-O staining showed cartilage fibrillation, clustering of chondrocytes, and a reduction in matrix proteoglycans at week 20 in the ACL-transected knee, but not in the sham-operated knee. Levels of glutamate and aspartate in dialysates from the ACL-transected knee were significantly increased by 92 +/- 20.3% or 57 +/- 17.5%, respectively, compared to those in the contralateral sham-operated knee. This increase may contribute to the pathogenesis of early OA. PMID- 15885478 TI - Cartilage degeneration in post-collapse cases of osteonecrosis of the human femoral head: altered mechanical properties in tension, compression, and shear. AB - Osteonecrosis, or avascular necrosis, is a painful and debilitating condition characterized by progressive joint degeneration subsequent to collapse of necrotic regions of trabecular bone. A clear understanding of the mechanism of cartilage degeneration in osteonecrosis is critical to the development of treatment strategies aimed at sparing the femoral head. An analysis of 13 post collapse osteonecrotic (ON) human femoral heads was performed relative to 24 non ON controls to determine quantitatively the biomechanical and histological properties of post-collapse osteonecrotic cartilage. Cartilage mechanical properties were measured in tension, compression, and shear at different sites on the femoral head and correlated to histologic measures of cartilage degeneration using a semi-quantitative grading scale. Decreasing cartilage tensile strength correlated with histologic evidence of degeneration in the ON group; however, less correlation was noted with shear and compressive properties. After statistical correction for the more severe histologic degeneration in the ON group, the non-ON samples were found to exhibit significantly smaller loss angles during shear testing, while samples from the ON group were found to have greater tensile strength. Similarly, the ON group was found to exhibit significantly greater proteoglycan loss while the non-ON group showed significantly increased surface fibrillation. This study provides evidence that the changes occurring in post-collapse cases of osteonecrosis may involve mechanisms other than those typically attributed to osteoarthritic degeneration. One potential explanation is that decreased loading of cartilage overlying collapsed ON lesions leads to proteoglycan loss similar to that occurring with cartilage disuse. Unlike degenerative changes, some articular cartilage changes caused by disuse may be reversible. The presence of reversible changes would likely increase the chances of success of head sparing treatment strategies that rely on the viability and mechanical integrity of the articular cartilage. PMID- 15885479 TI - Repair of porcine articular cartilage defect with autologous chondrocyte transplantation. AB - Articular cartilage is known to have poor healing capacity after injury. Autologous chondral grafting remains the mainstay to treat well-defined, full thickness, symptomatic cartilage defects. We demonstrated the utilization of gelatin microbeads to deliver autologous chondrocytes for in vivo cartilage generation. Chondrocytes were harvested from the left forelimbs of 12 Lee-Sung pigs. The cells were expanded in monolayer culture and then seeded onto gelatin microbeads or left in monolayer. Shortly before implantation, the cell-laden beads were mixed with collagen type I gel, while the cells in monolayer culture were collected and re-suspended in culture medium. Full-thickness cartilage defects were surgically created in the weight-bearing surface of the femoral condyles of both knees, covered by periosteal patches taken from proximal tibia, and sealed with a porcine fibrin glue. In total, 48 condyles were equally allotted to experimental, control, and null groups that were filled beneath the patch with chondrocyte-laden beads in gel, chondrocytes in plain medium solution, or nothing, respectively. The repair was examined 6 months post-surgery on the basis of macroscopic appearance, histological scores based on the International Cartilage Repair Society Scale, and the proportion of characteristic chondrocytes. Tensile stress-relaxation behavior was determined from uniaxial indentation tests. The experimental group scored higher than the control group in the categories of matrix nature, cell distribution pattern, and absence of mineralization, with similar surface smoothness. Both the experimental and control groups were superior to the null group in the above-mentioned categories. Viable cell populations were equal in all groups, but the proportion of characteristic chondrocytes was highest in the experimental group. Matrix stiffness was ranked as null > native cartilage > control > experimental group. Transplanted autologous chondrocytes survive and could yield hyaline-like cartilage. The application of beads and gel for transplantation helped to retain the transferred cells in situ and maintain a better chondrocyte phenotype. PMID- 15885480 TI - Treatment of cartilage with beta-aminopropionitrile accelerates subsequent collagen maturation and modulates integrative repair. AB - Integrative repair of cartilage was previously found to depend on collagen synthesis and maturation. beta-aminopropionitrile (BAPN) treatment, which irreversibly blocks lysyl oxidase, inhibited the formation of collagen crosslinks, prevented development of adhesive strength, and caused a buildup of GuHCl-extractable collagen crosslink precursors. This buildup of crosslink precursor in the tissue may be useful for enhancing integrative repair. We tested in vitro the hypothesis that pre-treatment of cartilage with BAPN, followed by washout before implantation, could be a useful therapeutic strategy to accelerate subsequent collagen maturation. In individual cartilage disks, collagen processing was reversibly blocked by BAPN treatment (0.1 mM) as indicated by a BAPN-induced increase in the total and proportion of incorporated radiolabel that was extractable by 4M guanidine-HCl, followed by a decrease, within 3-4 days of BAPN washout, in the proportion of extractable radiolabel to control levels. With a similar pattern, integration between pairs of apposed cartilage blocks was reversibly blocked by BAPN treatment, and followed by an enhancement of integration after BAPN washout. The low and high levels of integration were associated with enrichment in [(3)H]proline in a form that was susceptible and resistant, respectively, to extraction. With increasing duration up to 7 days after BAPN pre-treatment, the levels of [(3)H]proline extraction decreased, and the development of adhesive strength increased. Thus, BAPN can be used to modulate integrative cartilage repair. PMID- 15885481 TI - Repeated intraarticular injections of triamcinolone acetonide alter cartilage matrix metabolism measured by biomarkers in synovial fluid. AB - Although intraarticular (IA) corticosteroids are frequently used to treat joint disease, the effects of their repeated use on articular cartilage remains controversial. The aim of our study was to determine the effects of a clinically recommended dose of IA triamcinolone acetonide (TA), on synovial fluid (SF) biomarkers of cartilage metabolism. Ten adult horses, free of osteoarthritis (OA) in their radiocarpal joints, were studied. One radiocarpal joint of each horse was randomly chosen for treatment and the contralateral anatomically paired joint acted as the control. Aseptic arthrocentesis was performed weekly on both joints for 13 weeks. The initial results from the first 3 weeks of the experimental period established baseline untreated control marker levels for each joint, each being its own control. On weeks 3, 5, and 7, a sterile suspension of 12 mg of TA was injected into the treated joint and an equivalent volume of sterile saline solution (0.9%) was injected into the control joint. SF was immunoassayed for biomarkers of aggrecan turnover (CS 846 & KS), types I and II collagen cleavage (C1,2C) and type II collagen synthesis (CPII). In treated joints, there was a significant increase in CS 846, KS, C1,2C and CPII epitope concentrations following IA TA injections when compared to baseline levels. There was also a significant increase in C1,2C and CPII epitope concentrations in the contralateral control joints following IA TA injections in the treated joint. Significant differences were observed between treated and control joints for all markers except CPII. These findings indicate that TA alters articular cartilage and collagen metabolism in treated and, interestingly, also in control joints, suggesting a systemic effect of the drug. Though intuitively the observed findings would favor the hypothesis that long-term IA TA treatment changes joint metabolism and this may have detrimental effects; further studies would be necessary to confirm this. PMID- 15885482 TI - Bone morphogenetic protein 9 is a potent anabolic factor for juvenile bovine cartilage, but not adult cartilage. AB - Members of the bone morphogenetic protein (BMP) group of the TGF-beta superfamily have been shown to enhance matrix synthesis and maintain cartilage phenotype in long-term culture. These proteins have also been shown to augment cartilage repair in vivo, and may be of potential therapeutic benefit in the treatment of damaged articular cartilage. The present study was undertaken to examine the effects of BMP-9 on the metabolism of juvenile and adult bovine cartilage in vitro, and to compare the effects to those produced by two previously characterized BMPs: BMP-2 and 13 (CDMP-2). BMP-9 lead to a 7-8-fold stimulation of proteoglycan synthesis at the highest concentration tested, and a 6.4-fold stimulation of collagen synthesis at a concentration of 50 ng/mL in juvenile cartilage. BMP-2 also lead to a 7-8-fold increase in proteoglycan synthesis at the highest concentration tested, and was able to induce collagen synthesis 6.4 fold, but at a concentration of 1000 ng/mL. Proteoglycans isolated from BMP-9 treated cartilage exhibited an increased hydrodynamic size possibly due to increased glycosaminoglycan substitution or decreased C-terminal proteolysis. Consistent with the idea of limited C-terminal proteolysis, BMP-9 treatment lead to a significant reduction in the turnover rate of proteoglycans in juvenile explants. Interestingly, all three BMPs were unable to induce a measurable anabolic response in adult cartilage explants. PMID- 15885483 TI - Identification and characterization of valosin-containing protein (VCP/p97) in untransformed osteoblast-like cells. AB - A 97-kDa protein called valosin-containing protein (VCP) has been implicated in osteosarcoma metastasis and Paget's disease of bone, two conditions that complicate the course and outcome of orthopaedic surgery. High VCP gene expression is associated with high metastatic potential in osteosarcoma cells, while loss-of-function VCP mutations cause inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia (IBMPFD). VCP protein expression and regulation have not been examined in normal osteoblasts. The purpose of these studies was to characterize VCP protein expression in control and stressed untransformed osteoblasts. Proteins from confluent MC3T3-E1 mouse osteoblast-like cells were separated by 2D IEF/SDS-PAGE. An abundant spot with a M(r) of 94 kDa and a pI of 5.4 was identified as VCP by MALDI/ToF and peptide mass fingerprint analysis. High constitutive VCP protein expression in subconfluent and confluent resting and mildly physiologically stressed MC3T3-E1 cells was confirmed by Western blotting. When assessed by indirect immunofluorescence in fixed cells or Western blotting of subcellular fractions, VCP was more abundant in the cytoplasm than in the nucleus. Induction of mild physiological stress sufficient to stimulate the ubiquitin-proteasome pathway, which is partially dependent on VCP-mediated targeting of polyubiquitinylated substrates, did not affect steady-state VCP levels or distribution. Thus, VCP is a constitutively abundant protein in untransformed osteoblastic cells under all conditions tested. Such high levels of VCP protein expression in untransformed osteoblastic cells argue against a major causative role for it in metastasis, while the occurrence of Paget's disease in patients with missense VCP mutations supports a major role for VCP in normal osteoblast proliferation and regulation. PMID- 15885484 TI - Time for treating bone fracture using rhBMP-2: a randomised placebo controlled mouse fracture trial. AB - Although the mechanisms of osteoinduction by bone morphogenic proteins (BMPs) are increasingly understood, the most appropriate time to administer BMPs exogenously is yet to be clarified. The purpose of this study was to investigate when BMP may be administered to a fracture arena to maximise the enhancement of healing. Forty mice with externally fixed left femoral fractures were randomised into four groups: Group I, the control group was given a placebo of 30 microl saline at day 0; Groups II, III and IV were given 30 microl saline plus 2.5 microg rhBMP-2, at post-operative days 0, 4 or 8, respectively. Sequential radiographs were taken at days 0, 8, 16. On day 22 the mice were sacrificed and both femora were harvested for biomechanical assessment in 3-point bending and histological evaluation. Radiographic analysis indicated that healing of fractures in Groups II and III was significantly greater (p < 0.05) than those in Groups I and IV, at both 16 and 22 days post-fracture. The highest median bone mineral content at the fracture site was evidenced in Group III and II. Furthermore, Group III also had the highest relative ultimate load values, followed by Groups II, IV and I. Greater percentage peak loads were observed between Group I and both Groups II and III (p < 0.05). Histological examination confirmed that at 22 days post fracture, only fractures in Groups II and III had united with woven bone, and Groups I and IV still had considerable amounts of fibrous tissue and cartilage at the fracture gap. Data presented herein indicates that there is a time after fracture when rhBMP administration is most effective, and this may be at the time of surgery as well as in the early fracture healing phases. PMID- 15885485 TI - Prevention of atrophic nonunion development by recombinant human bone morphogenetic protein-7. AB - Severe periosteal and soft tissue disruption at the time of fracture may result in the formation of an atrophic nonunion. We have developed a reproducible atrophic nonunion in an animal model. The purpose of this study was to evaluate whether the immediate application of recombinant human BMP-7 to the fracture site could rescue the healing process in this nonunion model. A total of 56 three month old Fisher 344 rats were utilized. A 1.25 mm diameter K-wire was inserted into the femur in a retrograde fashion, and a mid-diaphyseal closed transverse fracture was created using a standard three point bending device. To create a nonunion, the fracture site was exposed and 2 mm of the periosteum was cauterized on each side of the fracture. The fracture site was immediately treated with either the application of rhBMP-7 50 microg in 25 microl of rat tail tendon collagen buffer (BMP-7 group), or with 25 microl of rat tail tendon collagen buffer only (Control group). Fracture healing was evaluated with serial radiographs every two weeks for an eight weeks period. Specimens at four and eight weeks were subjected to biomechanical and histological evaluation. None of the Control group healed throughout the eight weeks experimental duration. At four weeks 63% of the BMP-7 group had healed, and all had healed by six weeks. This investigation showed pronounced differences between the BMP-7 group and the Control group both histologically and biomechanically. In conclusion, we have demonstrated that the immediate application of BMP-7 may rescue the fracture healing process and prevent the development of nonunion following severe periosteal disruption. PMID- 15885486 TI - Initial vascularization and tissue differentiation are influenced by fixation stability. AB - Fracture healing requires a certain degree of mechanical stability and an adequate blood supply. The hypothesis of the present study was that increased interfragmentary shear leads to a reduced initial vascularization and prolonged healing. The aim of the study was to quantitatively analyze the histological appearance of vascularization and tissue differentiation with regard to fracture stability during the course of healing. A mid-shaft osteotomy of the tibia was performed in two groups of sheep and stabilized with either a rigid or semirigid external fixator, differing in bending stiffness. Interfragmentary movements and ground reaction forces were evaluated in vivo during a 9-week period. The sheep were sacrificed at 2, 3, 6, and 9 weeks postoperatively. The tibiae were tested biomechanically and histological sections from the callus were prepared for analysis of tissue differentiation and vascularization. Larger interfragmentary shear movements in the semirigid fixator group were associated with a reduced initial blood supply. At 6 weeks the semirigid fixator group showed a significantly lower percentage of mineralized bone and a higher amount of fibrous tissue leading to a significantly lower stiffness of the callus than the rigid fixator group. This initial delay in healing was compensated for in the later stages with the production of greater volumes of callus tissue so that both groups showed the same callus stiffness at 9 weeks. However, the rigid fixator group showed signs of the beginning of callus remodeling at the latest time points suggesting a faster bone healing. The results indicate the important role of the initial mechanical stability specifically in the vascularization of an osteosynthesis. Further studies should illustrate the precise role of mechanical conditions on the regulation of angiogenesis during early bone healing. PMID- 15885487 TI - Effect of low intensity pulsed ultrasound and BMP-2 on rat bone marrow stromal cell gene expression. AB - To determine how low intensity pulsed ultrasound alters gene expression in rat bone marrow stromal cells and to see if combining this stimulation with BMP-2, cells were pre-cultured for eight days in the presence of 50 microg/ml ascorbic acid and then exposed to either low intensity US or 100 ng/ml BMP-2 or both combined, beginning on the first, third fifth or seventh day of culture so that cells were exposed to the stimuli for one, three, five or seven days. Real time PCR was used to determine the effect of these treatments on gene expression of several genes associated with osteogenesis. The expression of some of the genes (Cbfa-1/Runx2, IGF-receptor, Alk-3, alkaline phosphatase, osteopontin, TGF-beta1, BMP-7) was increased compared to untreated controls. Combination of US and BMP-2 treatment did not lead to synergy of the two stimuli. Cbfa-1 stimulation occurred more quickly with US than with BMP-2. Increases in gene expression were greatest after 3 days exposure to US, with similar results for BMP-2 treatment implying that there may be a time dependence for the stimulus of osteogenic gene expression in stromal cells. PMID- 15885488 TI - Regulation of BMP-induced ectopic bone formation by Ahsg. AB - alpha2-HS-glycoprotein (Ahsg), also known as fetuin is a serum and bone resident glycoprotein, which binds to TGF-beta superfamily members including bone morphogenetic proteins (BMP) and inhibits dexamethasone-induced osteogenesis in bone marrow cultures in vitro. Here we demonstrate that Ahsg reduces cytokine binding to its cognate receptor in HOS osteocyte cells and suppresses intracellular signaling, while in vivo, we test the hypothesis that Ahsg deficient mice are hyper-responsive to BMP-induced osteogenesis. Human native BMP was implanted into the hindquarter muscles of Ahsg(+/+), Ahsg(+/-) and Ahsg(-/-) mice and 4 weeks later, ossicle formation was analyzed by radiography, bone density scanning (DEXA) and histomorphometry. Alkaline phosphatase (AP) activity was measured in ossicles as a marker for bone cell differentiation, and was significantly higher in Ahsg(-/-) versus Ahsg(+/-) and/or Ahsg(+/+) mice. Ectopic ossicle size in the Ahsg(+/-) mouse was 4-fold greater than that in the wild type (Ahsg(+/+)), and intermediate to that shown in Ahsg(-/-) mouse. Bone mineral density (BMD) was lower in the Ahsg(-/+) and Ahsg(-/-) mice compared to Ahsg(+/+) littermates. The ratio of cortical to cancellous bone was found to be >2-fold higher in Ahsg(-/-) mouse in comparison to the Ahsg(+/+) mice with no significant change in the Ahsg(-/+) mouse. Finally, a significantly higher incidence of satellite ossification; small islands of immature bone, was shown in Ahsg(-/-) mice as compared to Ahsg(+/+) mice. Although Ahsg binds to TGF-beta/BMP and blocks receptor signalling, it may also sequester cytokines in matrix, thereby acting as a reservoir of osteoinductive activity when released. This may explain the non-linear relationship between ectopic bone formation characteristics and Ahsg(+/+), Ahsg(+/-) and Ahsg(-/-) genotypes, although the increase in satellite bone formation might also explain this phenomenon. Our results suggest that Ahsg may be useful for prevention of the heterotopic ossification and the regulation of osteoinductive effects of BMP used with grafts. PMID- 15885489 TI - Mechanobiology of mandibular distraction osteogenesis: finite element analyses with a rat model. AB - Three-dimensional finite element (FE) analyses were performed to characterize the local mechanical environment created within the tissue regenerate during mandibular distraction osteogenesis (DO) in a rat model. Finite element models were created from three-dimensional computed tomography image data of rat hemi mandibles at four different time points during an optimal distraction osteogenesis protocol (i.e., most successful protocol for bone formation): end latency (post-operative day (POD) 5), distraction day 2 (POD 7), distraction day 5 (POD 10), and distraction day 8 (POD 13). A 0.25 mm distraction was simulated and the resulting hydrostatic stresses and maximum principal tensile strains were determined within the tissue regenerate. When compared to previous histological findings, finite element analyses showed that tensile strains up to 13% corresponded to regions of new bone formation and regions of periosteal hydrostatic pressure with magnitudes less than 17 kPa corresponded to locations of cartilage formation. Tensile strains within the center of the gap were much higher, leading us to conclude that tissue damage would occur there if the tissue was not compliant enough to withstand such high strains, and that this damage would trigger formation of new mesenchymal tissue. These data were consistent with histological evidence showing mesenchymal tissue present in the center of the gap throughout distraction. Finite element analyses performed at different time points during distraction were instrumental in determining the changes in hydrostatic stress and tensile strain fields throughout distraction, providing a mechanical environment rationale for the different levels of bone formation in end latency, and distraction day 2, 5, and 8 specimens. PMID- 15885490 TI - Thrombin peptide (TP508) promotes fracture repair by up-regulating inflammatory mediators, early growth factors, and increasing angiogenesis. AB - Previous studies have shown that a single injection of thrombin peptide (TP508) accelerates fracture repair in a closed rat femoral fracture model. The present study was conducted to elucidate the molecular mechanisms of TP508 action using Affymetrix genome-scale profiling and to link early gene expression changes to fracture histology and bone strength changes. Treatment of femoral fractures with TP508 accelerated fracture repair as determined by destructive torsion testing. Blinded histological analysis demonstrated that TP508-treated fracture callus had a significant increase in blood vessels relative to the controls. Gene array analysis showed that TP508 significantly induced expression of early growth factors, inflammatory response modifiers, and angiogenesis-related genes. This study therefore suggests that TP508 promotes fracture repair through a mechanism that involves an increased induction of a number of growth factors, enhanced expression of inflammatory mediators, and angiogenesis-related genes. PMID- 15885491 TI - rhBMP-2, rhVEGF(165), rhPTN and thrombin-related peptide, TP508 induce chemotaxis of human osteoblasts and microvascular endothelial cells. AB - Osteogenesis and angiogenesis are inter-linked and tightly regulated processes involved in growth, repair, and bone remodeling. Bone morphogenic protein 2 (BMP 2), vascular endothelial growth factor (VEGF), pleiotrophin (PTN) and thrombin related peptide, TP508 have all been found to have the ability to promote bone fracture healing by enhancing both the osteogenesis and angiogenesis processes. One of the underlying mechanisms proposed is that mediators for osteogenesis may also be involved in mediating angiogenesis and vice versa. The aim of this study was to examine the chemotactic effects of rhBMP-2, rhVEGF(165), rhPTN and TP508 on human osteoblasts and endothelial cells. Using a direct-viewing chemotaxis assay system, we report for the first time, the direct quantitative observation of chemotaxis of both human osteoblastc cells and microvascular endothelial cells towards sources of rhBMP-2, rhVEGF(165), rhPTN and TP508. This study confirmed that rhBMP-2, rhVEGF(165), rhPTN and TP508 have chemotactic effects on both human osteoblastic and endothelial cells, indicating that these factors are directly involved in promoting angiogenesis and osteogenesis by recruiting osteoblasts and endothelial cells via chemotaxis. PMID- 15885492 TI - Interfragmentary surface area as an index of comminution severity in cortical bone impact. AB - A monotonic relationship is expected between energy absorption and fracture surface area generation for brittle solids, based on fracture mechanics principles. It was hypothesized that this relationship is demonstrable in bone, to the point that on a continuous scale, comminuted fractures created with specific levels of energy delivery could be discriminated from one another. Using bovine cortical bone segments in conjunction with digital image analysis of CT fracture data, the surface area freed by controlled impact fracture events was measured. The results demonstrated a statistically significant (p < 0.0001) difference in measured de novo surface area between three specimen groups, over a range of input energies from 0.423 to 0.702 J/g. Local material properties were also incorporated into these measurements via CT Hounsfield intensities. This study confirms that comminution severity of bone fractures can indeed be measured on a continuous scale, based on energy absorption. This lays a foundation for similar assessments in human injuries. PMID- 15885493 TI - Comparison of gene expression measurements from cDNA and 60-mer oligonucleotide microarrays. AB - As the data generated by microarray technology continue to amass, it is necessary to compare and combine gene expression data from different platforms. To evaluate the performance of cDNA and long oligonucleotide (60-mer) arrays, we generated gene expression profiles for two cancer cell lines and compared the data between the two platforms. All 6182 unique genes represented on both platforms were included in the analysis. A limited correlation (r = 0.4708) was obtained and the difference in measurement of low-expression genes was considered to contribute to the limited correlation. Further restriction of the data set to differentially expressed genes detected in cDNA microarrays (1205 genes) and oligonucleotide arrays (1325 genes) showed modest correlations of 0.7076 and 0.6441 between the two platforms. Quantitative real-time PCR measurements of a set of 10 genes showed better correlation with oligonucleotide arrays. Our results demonstrate that there is substantial variation in the data generated from cDNA and 60-mer oligonucleotide arrays. Although general agreement was observed in measurements of differentially expressed genes, we suggest that data from different platforms could not be directly amassed. PMID- 15885494 TI - Biosilica formation in spicules of the sponge Suberites domuncula: synchronous expression of a gene cluster. AB - The formation of spicules is a complicated morphogenetic process in sponges (phylum Porifera). The primmorph system was used to demonstrate that in the demosponge Suberites domuncula the synthesis of the siliceous spicules starts intracellularly and is dependent on the concentration of silicic acid. To understand spicule formation, a cluster of genes was isolated. In the center of this cluster is the silicatein gene, which codes for the enzyme that synthesizes spicules. This gene is flanked by an ankyrin repeat gene at one side and by a tumor necrosis factor receptor-associated factor and a protein kinase gene at the other side. All genes are strongly expressed in primmorphs and intact animals after exposure to silicic acid, and this expression is restricted to those areas where the spicule formation starts or where spicules are maintained in the animals. Our observations suggest that in S. domuncula a coordinated expression of physically linked genes is essential for the synthesis of the major skeletal elements. PMID- 15885495 TI - Sexually dimorphic gene expression in the hypothalamus, pituitary gland, and cortex. AB - We examined sex differences in the transcriptomes of hypothalamus, pituitary gland, and cortex of male and female mice using serial analysis of gene expression. In total 940,669 tags were sequenced. In hypothalamus, 3 transcripts are differentially expressed by gender, including growth hormone (neuromodulation) and 3beta-hydroxysteroid dehydrogenase-1 (steroidogenesis). In pituitary gland, 43 transcripts are differentially expressed, including RAS guanyl-releasing protein 2 (cell signaling), ornithine transporter (mitochondrial transport), H3 histone family 3B (chromatin structure), heterogeneous nuclear ribonucleoprotein U (chromatin remodeling), NADH dehydrogenase (mitochondrial oxidative phosphorylation), neuronatin (cell differentiation), and ribosomal protein S27a (protein metabolism). EST X (inactive)-specific transcript antisense is expressed at a higher level in the three female organs, whereas growth hormone and NADH dehydrogenase are expressed at higher levels in female cortex. Thus, the current study has characterized key sexual dimorphisms in the transcriptomes of the hypothalamus, pituitary, and cortex. PMID- 15885496 TI - Origin of the prolactin-releasing hormone (PRLH) receptors: evidence of coevolution between PRLH and a redundant neuropeptide Y receptor during vertebrate evolution. AB - We present seven new vertebrate homologs of the prolactin-releasing hormone receptor (PRLHR) and show that these are found as two separate subtypes, PRLHR1 and PRLHR2. Analysis of a number of vertebrate sequences using phylogeny, pharmacology, and paralogon analysis indicates that the PRLHRs are likely to share a common ancestry with the neuropeptide Y (NPY) receptors. Moreover, a micromolar level of NPY was able to bind and inhibit completely the PRLH-evoked response in PRLHR1-expressing cells. We suggest that an ancestral PRLH peptide started coevolving with a redundant NPY binding receptor, which then became PRLHR, approximately 500 million years ago. The PRLHR1 subtype was shown to have a relatively high evolutionary rate compared to receptors with fixed peptide preference, which could indicate a drastic change in binding preference, thus supporting this hypothesis. This report suggests how gene duplication events can lead to novel peptide ligand/receptor interactions and hence spur the evolution of new physiological functions. PMID- 15885497 TI - A 700-kb physical and transcription map of the cervical cancer tumor suppressor gene locus on chromosome 11q13. AB - Nonrandom deletion of chromosome 11q13 sequences is a significant event in a number of human tumors. We have recently identified a 300-kb minimal area of deletion in primary cervical tumors that overlaps with deletions observed in endocrine and nasopharyngeal tumors. We have also observed a 5.7-kb homozygous deletion within this interval in HeLa cells (a cervical cancer cell line), HeLa cell-derived tumorigenic hybrids, and a primary cervical tumor, suggesting the presence of a tumor suppressor gene in this region. In the present investigation, we have constructed a 700-kb contig map encompassing the 300-kb deletion using the human genome sequence database and confirmed the map using various STS markers from the region. Our map also shows the overlap of a previously published rare, heritable fragile site, FRA11A, with the cervical cancer deletion locus. The mapped region contains highly repetitive GC-poor sequences. We have identified and characterized eight different polymorphic microsatellite markers from the sequences within and surrounding the deletion. Further, expression studies performed with 18 different ESTs localized adjacent to the homozygous deletion showed the presence of a transcript for only one of the ESTs, AA282789. This EST mapping within the homozygous deletion is also expressed in HeLa cells, thereby excluding the EST as the putative tumor suppressor gene. Additionally, analysis of four candidate genes (SF3B2, BRMS1, RIN1, and RAB1B) from the region showed expression of the expected size message in both the nontumorigenic and the tumorigenic HeLa cell hybrids, thereby excluding them as the putative tumor suppressor gene(s). However, Northern blot analysis with a fifth candidate gene, PACS1 (phosphofurin acidic cluster sorting protein), mapped to the deletion/FRA11A overlap region showed the expression of an 8-kb transcript in HeLa and five other tumor cell lines in addition to the expected 4.5-kb transcript. Since the gene shows abundant expression in normal tissues and an altered transcript is observed in tumor cell lines, we hypothesize that this gene could represent sequences of the putative tumor suppressor gene. Finally, we have observed a perfect 48-bp CAG/CCG repeat 99 kb proximal to D11S913, the marker linked to the neurodegenerative disorder spinocerebellar ataxia 5. The physical and transcription maps and the microsatellite markers of the 700-kb region of chromosome 11q13 should be helpful in the cloning of the cervical cancer tumor suppressor gene. PMID- 15885498 TI - Ovarian-specific expression of a new gene regulated by the goat PIS region and transcribed by a FOXL2 bidirectional promoter. AB - Studies on XX sex reversal in polled goats (PIS mutation: polled intersex syndrome) have led to the discovery of a female-specific locus crucial for ovarian differentiation. This genomic region is composed of at least two genes, FOXL2 and PISRT1, sharing a common transcriptional regulatory region, PIS. In this paper, we describe a third gene, PFOXic (promoter FOXL2 inverse complementary), located near FOXL2 in the opposite orientation. This gene composed of five exons encodes a 1723-bp cDNA, enclosing two repetitive elements in its 3' end. PFOXic mRNA encodes a putative protein of 163 amino acids with no homologies in any of the databases tested. The transcriptional expression of PFOXic is driven by a bidirectional promoter also enhancing FOXL2 transcription. In goats, PFOXic is expressed in developing ovaries, from 36 days postcoitum until adulthood. Ovarian-specific expression of PFOXic is regulated by the PIS region. PFOXic is found conserved only in Bovidae. But, a human gene located in the opposite orientation relative to FOXL2 can be considered a human PFOXic. Finally, we discuss evidence arguing for regulation of the level of FOXL2 transcription via the bidirectional promoter and the level of transcription of PFOXic. PMID- 15885499 TI - Gene expression profiling of dysplastic differentiation in cervical epithelial cells harboring human papillomavirus 16. AB - Molecular events occurring with high-risk human papillomavirus (HPV)-associated dysplastic differentiation of cervical epithelial cells are largely unknown. This study used differential display PCR to identify expression changes between nondifferentiating monolayer and differentiated organotypic (raft) cultures of W12 keratinocytes. These cells were originally derived from a clinical biopsy of HPV 16-positive dysplastic cervical epithelium and retain high-risk HPV 16 and the ability to differentiate, albeit with dysplastic morphology. Using this model system we identified 84 genes with changed expression during dysplastic differentiation. Most (70/84, approximately 80%) were down-regulated with differentiation, consistent with a restriction of expression during terminal differentiation. Twenty-two genes had no known function and 6 novel expressed sequence tags were identified among this group. Of the 62 genes with known functions, 25 belonged to transcription-, translation-, and posttranslation related categories and 30 had functions associated with neoplastic initiation/progression, calcium signaling, epithelial differentiation, and structure remodeling. Some of the genes with altered expression identified in this model of dysplastic differentiation may be useful biomarkers for early detection of cervical neoplasia and other HPV-associated oropharyngeal and anogenital cancers. PMID- 15885500 TI - Discovery of 342 putative new genes from the analysis of 5'-end-sequenced full length-enriched cDNA human transcripts. AB - In this work we describe the process that, starting with the production of human full-length-enriched cDNA libraries using the CAP-Trapper method, led us to the discovery of 342 putative new human genes. Twenty-three thousand full-length enriched clones, obtained from various cell lines and tissues in different developmental stages, were 5'-end sequenced, allowing the identification of a pool of 5300 unique cDNAs. By comparing these sequences to various human and vertebrate nucleotide databases we found that about 40% of our clones extended previously annotated 5' ends, 662 clones were likely to represent splice variants of known genes, and finally 342 clones remained unknown, with no or poor functional annotation. cDNA-microarray gene expression analysis showed that 260 of 342 unknown clones are expressed in at least one cell line and/or tissue. Further analysis of their sequences and the corresponding genomic locations allowed us to conclude that most of them represent potential novel genes, with only a small fraction having protein-coding potential. PMID- 15885501 TI - Expansion and diversification of KRAB zinc-finger genes within a cluster including Regulator of sex-limitation 1 and 2. AB - The genomic locus on mouse chromosome 13 called Regulator of sex-limitation (Rsl) accentuates sex differences in hepatic gene expression. Females homozygous for variant rsl alleles express some otherwise male-specific liver proteins, such as sex-limited protein (Slp), major urinary proteins (MUPs), and members of the cytochrome P450 (cyp) 2d subfamily. We recently identified mutations in two genes, Rsl1 and Rsl2, accounting for the rsl phenotype. These genes encode KRAB zinc-finger proteins (KRAB-ZFPs) and are embedded within a cluster of over 20 similar genes. Mammalian genomes contain over 200 KRAB-ZFP genes, which act biochemically to repress transcription, but the Rsl genes are the first to have their biological functions elucidated. Here we compare Rsl1 and Rsl2 with their neighboring genes, tracing a series of duplication, inversion, and gene conversion events that have created subfamilies within the locus. Polymorphisms among inbred mouse strains and feral species suggest that mutations responsible for the rsl phenotype arose during the creation of inbred strains. Comparisons among mouse, rat, and human sequences show that the Rsl genes, like members of certain other multigene families, have diversified in a species-specific manner. The targets of Rsl regulation also vary between species, occurring in gene families with functions in steroid and xenobiotic metabolism (Cyp2d), reproduction (MUPs), and immunity (Slp). This suggests that the Rsl locus in mouse, and comparable KRAB-ZFP genes in other mammals, may play a role in speciation via modulation of expression of genes influencing reproductive fitness or behavior. PMID- 15885502 TI - The WD-repeat protein GRWD1: potential roles in myeloid differentiation and ribosome biogenesis. AB - A cDNA fragment originally identified in U-937 cells as a vitamin D(3)-regulated gene is here designated the glutamate-rich WD-repeat (GRWD1) gene. WD-repeat proteins are a class of functionally divergent molecules that cooperate with other proteins to regulate cellular processes. GRWD1 encodes a 446-amino-acid protein containing a glutamate-rich region followed by four WD repeats. The yeast homologue of GRWD1, Rrb1, has been shown to be an essential protein involved in ribosome biogenesis. Northern analysis of GRWD1 message levels in the myeloid cell line HL-60 undergoing differentiation induced by vitamin D(3) or retinoic acid demonstrate downregulation coincident with slowing of cellular proliferation. A siRNA designed to downregulate GRWD1 similarly results in a decrease in cellular proliferation within 293 cells. Metabolic labeling of cells expressing the siRNA to GRWD1 shows a decrease in global protein synthesis. Finally, nuclear fractionation studies show cosedimentation of GRWD1 with preribosomal complexes, as well as the WD-repeat-containing protein Bop1, which has previously been implicated in ribosome biogenesis. These studies suggest that within mammalian cells GRWD1 plays a role in ribosome biogenesis and during myeloid differentiation its levels are regulated. PMID- 15885503 TI - In vivo characterization of a vertebrate ultraconserved enhancer. AB - Genomic sequence comparisons among human, mouse, and pufferfish (Takifugu rubripes (Fugu)) have revealed a set of extremely conserved noncoding sequences. While this high degree of sequence conservation suggests severe evolutionary constraint and predicts a lack of tolerance to change to retain in vivo functionality, such elements have been minimally explored experimentally. In this study, we describe the in-depth characterization of an ancient conserved enhancer, Dc2, located near the dachshund gene, which displays a human-Fugu identity of 84% over 424 basepairs (bp). In addition to this large overall conservation, we find that Dc2 is characterized by the presence of a large block of sequence (144 bp) that is completely identical among human, mouse, chicken, zebrafish, and Fugu. Through the testing of reporter vector constructs in transgenic mice, we observed that the 424-bp Dc2-conserved element is necessary and sufficient for brain tissue enhancer activity. In vivo analyses also revealed that the 144-bp 100% conserved sequence is necessary, but not sufficient, to replicate Dc2 enhancer function. However, the introduction of two separate 16-bp insertions into the highly conserved enhancer core did not cause any detectable modification of its in vivo activity. Our observations indicate that the 144-bp 100% conserved element is tolerant of change at least at the resolution of this transgenic mouse assay and suggest that purifying selection on the Dc2 sequence might not be as strong as we predicted or that some unknown property also constrains this highly conserved enhancer sequence. PMID- 15885504 TI - A conserved noncoding intronic transcript at the mouse Dnm3 locus. AB - A 6-kb antisense transcript (Dnm3os) contained within an intron of the mouse Dnm3 gene has been identified in a screen for genes that may be regulated by the basic helix-loop-helix transcription factor Twist during mouse development. The antisense transcript is highly conserved between vertebrate species, but does not appear to encode a protein. We show that expression of the Dnm3 and Dnm3os transcripts overlaps during embryogenesis and in adult tissues, except that Dnm3 is most highly expressed in adult brain and testis and expressed at lower levels in embryos, whereas the antisense transcript is most strongly expressed in embryos and gravid uterus. Both Dnm3 and Dnm3os are downregulated in branchial arch tissue of Twist-null embryos. The conservation and restricted expression pattern of this noncoding transcript suggest that it may perform an important function during embryonic development and further suggest a connection between the regulation of Dnm3 and this novel intronic, antisense transcript. PMID- 15885505 TI - High-resolution array CGH increases heterogeneity tolerance in the analysis of clinical samples. AB - Recent advances in array comparative genomic hybridization (array CGH) technology are revolutionizing our understanding of tumor genomes. Marker-based arrays enable rapid survey at megabase intervals, while tiling path arrays examine the entire genome in unprecedented detail. Tumor biopsies are typically small and contain infiltrating stromal cells, requiring tedious microdissection. Tissue heterogeneity is a major barrier to high-throughput profiling of tumor genomes and is also an important consideration for the introduction of array CGH to clinical settings. We propose that increasing array resolution will enhance detection sensitivity in mixed tissues and as a result significantly reduce microdissection requirements. In this study, we first simulated normal cell contamination to determine the heterogeneity tolerance of array CGH and then validated this detection sensitivity model on cancer specimens using the newly developed submegabase resolution tiling-set (SMRT) array, which spans the human genome with 32,433 overlapping BAC clones. PMID- 15885506 TI - Is unawareness of psychotic disorder a neurocognitive or psychological defensiveness problem? AB - We examined whether deficits in attention and perceptual encoding as well as psychological defensiveness were associated with impaired awareness of disorder in schizophrenia. The Scale for Unawareness of Mental Disorder (SUMD) was administered to 52 outpatients with a recent onset of schizophrenia approximately 1-2 months following hospital discharge. Two versions of the Continuous Performance Test (CPT) were used to measure attentional impairment--the Degraded Stimulus CPT (DS-CPT) and a memory-load version (3-7 CPT). Three scales from the Minnesota Multiphasic Personality Inventory were used as indicators of psychological defensiveness: Scales L (Lie), K (Correction), and R (Repression). The Classification and Regression Tree (CART) program, a nonparametric statistical method, was used to identify relationships among multiple predictor variables and to provide optimal splitting scores for each predictor variable. Different combinations of poor target discrimination (d') on the 3-7 CPT and a cautious response style on the DS-CPT were associated with the three levels of overall unawareness of having a mental disorder. For nonpsychotic patients, better target discrimination (d') on the 3-7 CPT tended to be associated with better awareness of having a mental disorder. In contrast, unawareness among the patients who were psychotic at the time of the SUMD administration was not discriminated by attentional measures, but was associated with a combination of two measures of psychological defensiveness from the MMPI reflecting guardedness, psychological suppression, attempting to present oneself in a socially desirable light, and social acquiescence. Generally similar associations were found for two other dimensions of poor insight: unawareness of the beneficial effects of antipsychotic medication, and inability to attribute unusual thoughts and hallucinatory experiences to a mental disorder. PMID- 15885507 TI - Attention orienting dysfunction during salient novel stimulus processing in schizophrenia. AB - Schizophrenia is characterised by marked disturbances of attention and information processing. Patients experience difficulty focusing on relevant cues and avoiding distraction by irrelevant stimuli. Event-related potential recordings indicate an amplitude reduction in the P3a component elicited by involuntary orienting to task-irrelevant, infrequent novel stimuli presented during auditory oddball detection in patients with schizophrenia. The goal of the present study was to elucidate the functional abnormality underlying the disturbed orienting to novel stimuli in schizophrenia. Twenty-eight stable, partially remitted, medicated patients with schizophrenia and 28 healthy control participants completed a novelty oddball variant during event-related fMRI. Relative to healthy participants, patients with schizophrenia were characterised by underactivity during novel stimulus processing in the right amygdala hippocampus, within paralimbic cortex in the rostral anterior cingulate and posterior cingulate cortices and the right frontal operculum, and in association cortex at the right temporo-parietal-occipital junction, bilateral intraparietal sulcus, and bilateral dorsal frontal cortex. Subcortically, relative hypoactivation during novelty processing was apparent in the cerebellum, thalamus, and basal ganglia. These results suggest that patients less efficiently reorient processing resources away from the ongoing task of detecting and responding to the task-relevant target stimuli. In addition, trend results suggest that patients experienced increased distraction by novel stimuli. PMID- 15885508 TI - The implications of memory profiles in schizophrenia on vocational and neuropsychological functioning. AB - Studies in schizophrenia suggest that verbal learning and memory may distinguish three subgroups of patients: an unimpaired memory profile group and two groups that have memory profiles similar to those seen in cortical and subcortical dementias. Using the Hopkins Verbal Learning Test, Revised edition (HVLT-R), this study attempted to differentiate patients into three memory profile groups and to examine the validity of these groups with respect to vocational outcomes and neuropsychological functioning. Results from this study replicated previous findings and extended them by demonstrating a link to vocational outcome. In addition, the proportion of patients in each group closely resembled that obtained in previous studies. Specifically, the relatively unimpaired memory group (42%) showed overall better memory and neuropsychological performance than the two impaired groups, the subcortical group (38%) showed impaired recall but intact recognition and deficits in visuospatial functioning, and the cortical group (20%) showed deficits in recall, recognition, and sustained attention/executive functioning. There were no clinical differences between the three groups, but both the unimpaired and subcortical groups increased the number of hours worked following a vocational rehabilitation program. Given these differences, more research is warranted to explore the effect of memory impairment subtypes on vocational outcome measures. PMID- 15885509 TI - The course and clinical correlates of dysfunctions in visual perceptual organization in schizophrenia during the remission of psychotic symptoms. AB - This study evaluated deficits in visual perceptual organization in schizophrenia over the course of inpatient treatment, in relation to the remission of particular psychotic symptoms. Disorganized (n=14) and non-disorganized (n=33) schizophrenia patients were tested upon admission to an inpatient psychiatric unit, and again after 3 weeks of treatment, on two measures of visual perceptual organization. Performance of schizophrenia patients was compared to groups of patients with psychotic disorders other than schizophrenia (n=19) and non psychotic psychiatric disorders (n=25). Symptom ratings were collected at both assessment points. Deficits in visual perceptual organization were observed for both tasks in disorganized schizophrenia patients at index and these deficits improved during the course of treatment. Moreover, improvement in visual perceptual organization correlated significantly with reductions in disorganized symptoms in the schizophrenia group. We interpret these data as further support for the hypothesis that the disorganization syndrome in schizophrenia reflects a widespread deficit in the cognitive coordination of contextually related stimuli, leading to dysfunctional organization of stimulus features in vision, thought and language. PMID- 15885510 TI - A preliminary study of interactive questioning methods to assess and improve understanding of informed consent among patients with schizophrenia. AB - Growing recognition of the inadequacy of traditional methods of providing informed consent, especially for individuals vulnerable to impaired decisional capacity, has spurred recent interest in how to assess and improve components of consent-related decision making. In this preliminary study, we aimed to compare different methods of interactive questioning during presentation of research consent information among patients with schizophrenia. Patients were randomized to receive either standard administration (SA) of a consent form or one of two interactive questioning methods: Corrective Feedback (CF), in which the correct answer was provided following the participant's response, or Errorless Learning (EL), in which correct answers were provided just prior to the question. The MacArthur Competence Assessment Tool for Clinical Research (MacCAT-CR) was used to measure understanding, appreciation, reasoning, and expression of a choice following presentation of the consent form. There was no significant effect of condition (SA vs. EL vs. CF) on any of the components of decisional capacity. Understanding scores measured during the consent process were higher than those measured afterward, but the two scores were highly correlated. Thus, the results of this randomized study suggest that interactive questioning neither helped nor harmed understanding, appreciation, or reasoning among patients with schizophrenia. Other considerations, however, may favor use of such methods in the consenting process. PMID- 15885511 TI - Retrieval-induced forgetting in schizophrenia. AB - Retrieving category associates (e.g., FRUIT-ORANGE) may induce forgetting other category members (e.g., FRUIT-BANANA), a phenomenon known as retrieval-induced forgetting (RIF). We designed 2 experiments to examine the role of RIF in the associative memory impairment of schizophrenia (SZ). Subjects studied 36 category exemplar pairs, generated from 6 categories composed of 6 members each. For half of the studied category-exemplar pairs, subjects practiced retrieval by completing word stems, followed by a delayed category-cued recall on all of the practiced and unpracticed items. Experiment 1 used unrelated category exemplars pairs (e.g., FRUIT-ORANGE, METALS-IRON), whereas experiment 2 included related category exemplar pairs (e.g., COTTON-SHIRT, LEATHER-SKIRT). SZ showed reduced associative memory but normal RIF for unrelated categories used in experiment 1. For experiment 2, SZ showed a significant decline in associative memory for related but not unrelated category-exemplars in comparison to controls. Results suggested faulty specificity/distinctiveness for encoding and retrieval, but not abnormal RIF in the associative memory disturbance of SZ. PMID- 15885512 TI - Relationship of impaired processing speed and flexibility of abstract thought to improvements in work performance over time in schizophrenia. AB - Vocational impairments in schizophrenia have been widely linked to deficits in neurocognition. This study examined the possibility that deficits in visuomotor processing speed and flexibility in abstract thought may in combination be an especially potent risk factor for poorer levels of work performance in rehabilitation. Fifty-seven participants with confirmed diagnoses of schizophrenia spectrum disorders were administered the Digit Symbol Subtest and the Wisconsin Card Sorting Test and then offered work placements in a vocational rehabilitation program. Work performance was assessed biweekly over two months using the Work Behavior Inventory. Multivariate and univariate repeated ANOVA revealed that participants classified as having no impairments in either visuomotor processing speed and flexibility in abstract thought (n=14) had superior work performance compared to participants with deficits in either one area (n=20) or both areas (n=23). Additionally, participants with no impairments were the only group to show significant improvement in work performance over 7 weeks of rehabilitation. Participants with only one deficit had significantly better work performance than participants with both deficits but showed no statistically significant improvement in work performance. Results suggest assessments of these domains of neurocognition may provide important information about individual needs for adjunct services. PMID- 15885513 TI - Computerized cognitive remediation improves verbal learning and processing speed in schizophrenia. AB - Computerized cognitive remediation has resulted in improved executive function in schizophrenia, whereas results with regard to verbal memory were inconsistent. In the present study, 42 inpatients with schizophrenia were randomly assigned to a computerized cognitive remediation group or to a treatment-as-usual (TAU) control group. The remediation group received 15 sessions of computerized cognitive training (Cogpack) over a 3-week period. Neurocognitive functions were assessed at the beginning and end of this period. Compared to the control condition, remediation training resulted in improvements in verbal learning, processing speed and executive function (verbal fluency). The results indicate that cognitive remediation may lead to improvements beyond those of executive function. PMID- 15885514 TI - Thought disorder, pragmatic language impairment, and generalized cognitive decline in schizophrenia. AB - BACKGROUND: Schizophrenia is associated with pragmatic language impairment (PLI), a reduced ability to communicate intention in a rule-governed fashion. Two explanations for PLI include that PLI is equivalent to thought disorder and that PLI is secondary to generalized cognitive decline. OBJECTIVES: The aims of this study were to demonstrate PLI in schizophrenia and to test which of these explanations best accounts for the relationships among thought disorder, PLI, and generalized cognitive decline. METHOD: Schizophrenia (n=20) and control (n=26) participants provided speech samples that were scored for thought disorder (type token ratio and Cloze procedure) and PLI [Profile of Pragmatic Impairment in Communication (PPIC)]. Generalized cognitive decline was determined from discrepancies between current and premorbid verbal IQ. RESULTS: Patients with schizophrenia exhibited significant PLI and generalized cognitive decline. There was no evidence of an association between thought disorder and PLI. Moreover, generalized cognitive decline predicted PLI (r(2)=0.33 to 0.59) but not thought disorder (r(2)=0.02 to 0.06). CONCLUSIONS: The results conformed to a predicted pattern of associations based on the notion that PLI in schizophrenia is secondary to generalized cognitive decline. PMID- 15885515 TI - Proverb comprehension reconsidered--'theory of mind' and the pragmatic use of language in schizophrenia. AB - BACKGROUND: For decades, impaired proverb comprehension has been regarded as typical of schizophrenic thought disorder. Testing patients' proverb understanding has widely been abandoned, however, due to poor reliability and validity of the assessment procedures. Since the underlying cognitive deficit of impaired proverb interpretation remained obscure, this study sought to determine the relation of proverb understanding with other cognitive domains, particularly 'theory of mind' or 'mindreading', in schizophrenia. METHODS: 31 patients diagnosed with schizophrenia were assessed using a novel German Proverb Test [Barth, A., Kufferle, B., 2001. Die Entwicklung eines Sprichworttests zur Erfassung konkretistischer Denkstorungen bei schizophrenen Patienten. Nervenarzt 72, 853-858.], a 'theory of mind' test battery, a variety of executive functioning tests and verbal intelligence. Psychopathology was measured using the PANSS [Kay, S.R., Opler, L.A., Lindenmayer, J.P., 1989. The Positive and Negative Syndrome Scale (PANSS): rationale and standardisation. Br. J. Psychiatry 158 (suppl. 7), 59-67.]. Patients' task performance was compared to a group of healthy control persons. RESULTS: 'Theory of mind', executive functioning and intelligence were strongly correlated with patients' ability to interpret proverbs correctly. In a regression analysis 'theory of mind' performance predicted, conservatively estimated, about 39% of the variance of proverb comprehension in the patient group. CONCLUSIONS: The ability to interpret such metaphorical speech that is typical of many proverbs crucially depends on schizophrenic patients' ability to infer mental states. Future studies may further address differences between schizophrenia subtypes or the relation to specific symptom clusters. PMID- 15885516 TI - Language reactivity and work functioning in schizophrenia. AB - Some studies have found that the speech of certain schizophrenia patients becomes more disordered in stressful laboratory situations. It is unknown, however, whether affective reactivity of speech is associated with stress responsiveness of symptoms in the real world. This study examines whether language-reactive patients report more stress-related impairments in work functioning than language nonreactive patients. Forty-six patients provided speech samples and completed a work history interview. It was found that the language-reactive patients were more likely than the language-nonreactive patients to endorse items pertaining to social anxiety and difficulty relating to others as reasons for their work difficulties. This suggests that language-reactive patients are more sensitive to social stressors than language-nonreactive patients. PMID- 15885517 TI - A 4.0-T fMRI study of brain connectivity during word fluency in first-episode schizophrenia. AB - OBJECTIVE: To use functional magnetic resonance imaging (fMRI) to investigate functional connectivity, and hence, underlying neural networks, in never-treated, first-episode patients with schizophrenia using a word fluency paradigm known to activate prefrontal, anterior cingulate, and thalamic regions. Abnormal connectivity between the prefrontal cortex (PFC) and other brain regions has been demonstrated in chronic, medicated patients in previous positron emission tomography (PET) studies, but has not to our knowledge, previously been demonstrated using both first-episode, drug-naive patients and fMRI technology. METHODS: A 4.0-Tesla (T) fMRI was used to examine activation and functional connectivity [psychophysiological interactions (PPIs)] during a word fluency task compared to silent reading in 10 never-treated, first-episode patients with schizophrenia and 10 healthy volunteers of comparable age, sex, handedness, and parental education. RESULTS: Compared to healthy volunteers, the schizophrenia patient group exhibited less activation during the word fluency task, mostly in the right anterior cingulate and prefrontal regions. Psychophysiological interactions between right anterior cingulate and other parts of the brain revealed a localized interaction with the left temporal lobe in healthy volunteers during the task and a widespread unfocussed interaction in patients. CONCLUSION: These findings suggest anterior cingulate involvement in the neuronal circuitry underlying schizophrenia. PMID- 15885518 TI - Correlations between MRI-assessed volumes of the thalamus and cortical Brodmann's areas in schizophrenia. AB - BACKGROUND: We compared the thalamic-cortical volumetric correlational patterns in patients with schizophrenia and normal comparison subjects, and evaluated their relations to outcome. METHODS: High-resolution MR images were acquired in patients with schizophrenia (n=106) and normal comparison subjects (n=42). Patients were divided into good-outcome (n=52) and poor-outcome (Kraepelinian, n=54) subtypes based on their ability for self-care. Correlations between the relative gray and white matter volumes of the individual cortical Brodmann's areas and five dorsoventral levels of the thalamus were assessed. RESULTS: Compared to normal subjects, schizophrenia patients lacked significant thalamic gray matter volume correlations with the prefrontal and medial temporal cortical regions in the right hemisphere, and with frontal, cingulate, posterior parietal and occipital regions in the left hemisphere, while normal white matter volume cortical-thalamic correlations along the cingulate gyrus and in the temporal lobe were not found in schizophrenia patients in both hemispheres. In contrast to both normal comparison subjects and good-outcome group, schizophrenia patients with poor outcomes showed significant bilateral gray matter volume correlations between the dorsal thalamus and ventral prefrontal cortex, while the group differences in the white matter volume correlations were mostly restricted to the cingulate arch. CONCLUSIONS: Whereas patients with schizophrenia exhibit deficiencies in cortical-thalamic correlational patterns, poor outcome is associated with abnormal interregional correlations not observed in either normal subjects or patients with good outcomes. This latter finding may be explained by a core neurodevelopmental disturbance that results in aberrant cortical-thalamic connectivity in poor-outcome schizophrenia. PMID- 15885519 TI - Hippocampus and amygdala volumes in schizophrenia and other psychoses in the Northern Finland 1966 birth cohort. AB - Structural brain differences have been reported in many studies with schizophrenia, but few have involved a general population birth cohort. We investigated differences in volume, shape and laterality of hippocampus and amygdala in patients with schizophrenia, all psychoses and comparison subjects within a large general birth cohort sample, and explored effects of family history of psychosis, perinatal risk and age-at-onset of illness. All subjects with psychosis from the Northern Finland 1966 birth cohort were invited to a survey including MRI scan of the brain, conducted in 1999-2001. Comparison subjects not known to have psychosis were randomly selected from the same cohort. Volumes of hippocampus and amygdala were measured in 56 subjects with DSM-III-R schizophrenia, 26 patients with other psychoses and 104 comparison subjects. Small hippocampal volume reductions in schizophrenia (2%) and all psychoses (3%) were not significant when adjusted for total brain volume. The shape of hippocampus in schizophrenia did not differ significantly from comparison subjects. Right hippocampus and amygdala were significantly larger than the left in all groups. Mean amygdala volume in schizophrenia or all psychoses did not differ from comparison subjects. Patients with family history of psychosis had larger hippocampus than patients without. Neither perinatal risk nor age-at-onset of illness had any effect on hippocampal or amygdala volumes. Small hippocampal volume reduction in schizophrenia and all psychoses was not disproportionate to reduced whole brain volume in this population-based sample. Perinatal events that have been suggested as of etiological importance in structural pathology of psychosis had no effect. PMID- 15885520 TI - Hippocampal and anterior cingulate morphology in subjects at ultra-high-risk for psychosis: the role of family history of psychotic illness. AB - BACKGROUND: While structural brain imaging abnormalities have been identified in schizophrenia and related disorders, it is unclear when they arise. Some appear to predate the illness and may be genetic in origin, while others are associated with the onset of the disorder. METHODS: We examined the hippocampal volumes and anterior cingulate morphology from the MRI scans of 79 male subjects at ultra high-risk (UHR) for developing psychosis, 35 of whom had a family history of schizophrenia, and compared them with 49 healthy male volunteers. RESULTS: Analysis of covariance demonstrated that left hippocampal volumes were significantly smaller in the UHR group without a family history of schizophrenia, when compared to the UHR group with such history. A similar pattern was found for the left anterior cingulate region, both in terms of reduced paracingulate folding and cingulate sulcus interruptions, although this did not reach significance. CONCLUSIONS: We found that a family history of schizophrenia was not associated with a greater degree of structural brain abnormalities in an ultra-high-risk group, and in fact it was those UHR patients without such history who displayed greater abnormalities, although this only reached significance for the left hippocampus. Thus, it appears that the mechanisms that result in gross morphological anomalies in the hippocampus and anterior cingulate in psychosis are driven more by environmental than genetic factors. PMID- 15885521 TI - Increased N-acetylaspartate in rat striatum following long-term administration of haloperidol. AB - N-acetylaspartate (NAA) is present in high concentrations in the CNS and is found primarily in neurons. NAA is considered to be a marker of neuronal viability. Numerous magnetic resonance spectroscopy (MRS) and postmortem studies have shown reductions of NAA in different brain regions in schizophrenia. Most of these studies involved patients chronically treated with antipsychotic drugs. However, the effect of chronic antipsychotic treatment on NAA remains unclear. In the present study, we measured NAA in brain tissue taken from 43 male Long-Evans rats receiving 28.5 mg/kg haloperidol decanoate i.m. every 3 weeks for 24 weeks and from 21 controls administered with vehicle. Determination of tissue concentrations of NAA was achieved by HPLC of sections of frozen tissue from several brain regions with relevance to schizophrenia. Chronic administration of haloperidol was associated with a significant increase (+23%) in NAA in the striatum (p<0.05) when compared to controls, with no significant changes in the other regions investigated (frontal and temporal cortex, thalamus, hippocampus, amygdala, and nucleus accumbens). NAA appears to be selectively increased in the striatum of rats chronically receiving haloperidol. This increase may reflect a hyperfunction of striatal neurons and relate to the reported increase in somal size of these cells and/or the increase in synaptic density seen in this region following antipsychotic administration. The lack of effect in other regions indicates that the well-documented NAA deficits seen in chronically treated schizophrenia patients is not an effect of antipsychotic medication and may in fact be related to the disease process. PMID- 15885522 TI - Reduced cortical folding in individuals at high risk for schizophrenia: a pilot study. AB - While cortical gyrification anomalies have been reported in schizophrenia, it is unknown if individuals at high risk for schizophrenia (HR) might also exhibit abnormal cortical folding. Using MRI scans, the gyrification index (GI) was calculated for 9 adolescent HR males and 12 healthy male controls. Using the first coronal slice anterior to the corpus callosum, cortical contours were manually traced to calculate the GI. The left GI was lower in HR when compared to controls, but no difference in the right GI was observed. These results are consistent with studies of affected individuals, supporting genetic and neurodevelopmental models of schizophrenia. PMID- 15885523 TI - Risk for cancer in a cohort of patients hospitalized for schizophrenia in Denmark, 1969-1993. AB - We investigated the cancer risk of patients hospitalized for schizophrenia in a nationwide cohort study. All 22766 adults admitted for schizophrenia, ICD-8 295, in Denmark between 1969 and 1993 were followed up for cancer through 1995. The incidence of site-specific cancers was compared with national incidence rates, adjusted for sex, age and calendar time. The risk for cancer was increased for both men and women during the first year of follow-up. When the first year of follow-up was excluded, the risk for all tobacco-associated cancers and for prostate and rectal cancers was reduced for male patients with schizophrenia. The standardized incidence ratio (SIR) of lung cancer was marginally reduced (SIR, 0.86; 95% CI: 0.65, 1.02) for male patients with schizophrenia; this was due, however, to a reduction in risk for older patients. An increased risk for breast cancer found for female patients with schizophrenia (SIR, 1.20; 95% CI: 1.05, 1.38) should be interpreted with caution, given the high proportion of nulliparous women with schizophrenia in Denmark. The data might support reduced risks for prostate and rectal cancer among male patients with schizophrenia, whereas a changing smoking pattern might explain the reduced risk for tobacco related cancers. PMID- 15885524 TI - Effects of chronic typical and atypical antipsychotic drug treatment on maternal behavior in rats. AB - Understanding the effects of antipsychotics on maternal behavior is important for understanding the poor quality of mother-infant interaction in schizophrenia. Previous preclinical work has demonstrated that acute treatment with typical and atypical antipsychotics disrupts maternal behavior. However, the effects of chronic antipsychotic treatment on maternal behavior are unknown. This issue is of importance since clinical use of antipsychotic medication requires continuous exposure to these drugs. In this study, we treated postpartum rats with haloperidol (0.25 mg/kg/day) or olanzapine (7.5 mg/kg/day), via osmotic minipumps or daily injections for 3 weeks. Maternal behavior was assessed every third day. On each observation day, maternal behavior was observed twice, once just prior to the daily injection ("trough" as this was 24 h after last injection) and again 2 h after the injection ("peak"). Daily injections of haloperidol and olanzapine significantly disrupted pup retrieval, pup licking, nest building at peak, but this effect was gone by trough. Drug administration via minipumps also disrupted these behaviors, but the effects were less severe. Pup nursing was enhanced by either method of drug administration. No evidence of sensitization or tolerance associated with chronic drug treatment was found. It is concluded that chronic antipsychotic treatment disrupts active maternal behaviors and this disruption, most likely to lead to side effects in humans, should be avoided in future drug development. PMID- 15885525 TI - Three-year recovery outcomes for long-term patients with co-occurring schizophrenic and substance use disorders. AB - Little is known about the expected treatment outcomes of patients with co occurring schizophrenic and substance use disorders. This paper reports 3-year outcomes for 152 patients with schizophrenia or schizoaffective disorder and substance use disorders, all of whom received integrated dual disorders treatments in the New Hampshire Dual Diagnosis Study. Outcomes are defined as positive coping behaviors identified by consumers as indicators of recovery. Participants improved steadily in terms of controlling symptoms of schizophrenia, actively attaining remissions from substance abuse, increasing competitive employment, increasing social contacts with non-substance abusers, and improving life satisfaction. Though successful in reducing hospitalization and homelessness, they did not increase time in independent living situations. Outcomes were only weakly interrelated, suggesting that recovery is a multidimensional concept. Neither psychotic diagnosis (schizophrenia vs. schizoaffective disorder) nor substance abuse diagnosis (alcohol vs. other drug disorder vs. both) was related to outcomes. However, these patients with co occurring schizophrenic and substance use disorders did significantly less well than patients with co-occurring bipolar and substance use disorders in terms of hospitalization, independent living, and quality of life. Overall, the findings provide a hopeful long-term perspective for dual diagnosis patients. PMID- 15885526 TI - Obsessive-compulsive symptoms in schizophrenia: associated clinical features, cognitive function and medication status. AB - OBJECTIVE: To determine the prevalence and clinical significance of obsessive compulsive (OC) symptoms among a group of stable outpatients with schizophrenia. METHODS: We studied 118 patients with schizophrenia from an urban clinic, characterized using clinical symptoms scales, including the Yale-Brown Obsessive Compulsive Scale (Y-BOCS), and neuropsychological testing. We categorized patients into three groups according to severity of OC symptoms and used multivariate linear regression and chi-square tests to compare groups on variables of interest. RESULTS: Only 10 patients (8.8%) had Y-BOCS scores greater than 16, a standard criterion for OCD studies. The patient group with the most OC symptoms (Y-BOCS scores >11) scored higher on the Hamilton Depression Scale, the positive symptoms subscale of the Positive and Negative Syndromes Scale (PANSS) and its delusions item, but not on any of the neuropsychological tests compared to the other two groups. Patients with most severe compulsive symptoms (but not OC symptoms together, or obsessions alone) were more likely to be treated with olanzapine or clozapine, atypical antipsychotic medications previously reported to induce or worsen OC symptoms. CONCLUSIONS: Our results confirm previous findings that patients with schizophrenia and comorbid OC symptoms have more positive symptoms but not the suggestion that such patients are more cognitively impaired than their counterparts without OC symptoms. We suggest possible explanations for discrepancies in the literature, including differences in patient sampling and definition of comorbid OC symptoms. Finally, our data suggest that olanzapine and clozapine may produce or worsen compulsions in some patients; prospective studies need to address this possibility. PMID- 15885527 TI - Stressful life events in recent-onset schizophrenia: reduced frequencies and altered subjective appraisals. AB - While research strongly supports the notion that stressful life events may trigger the exacerbation of psychotic symptoms in schizophrenia, the mechanisms through which affected individuals respond to life events during the early course of this disorder have received limited attention. This 12-month longitudinal study compared the frequencies, qualitative characteristics, and subjective appraisals of life events in recent-onset schizophrenia patients (n=78) and a nonpatient comparison sample (n=63). Negative and positive life events were assessed using a semi-structured interview every 4 weeks among patients and approximately every 4 months among controls, and participants appraised each event they experienced in terms of emotional impact, controllability, and effectiveness in handling the event. Schizophrenia patients reported significantly lower rates of life events than their nonpsychiatric counterparts across nearly every type of negative and positive event examined. In the context of generally lower event frequencies, patients appraised the negative and positive events they did experience as less controllable and more poorly handled than controls, and also appraised positive events as less desirable. Results are discussed in terms of their implications for understanding susceptibility to stress during the early course of schizophrenia. PMID- 15885528 TI - The psychotic spectrum: validity and reliability of the Structured Clinical Interview for the Psychotic Spectrum. AB - This study evaluates the validity and the reliability of a new instrument developed to assess the psychotic spectrum: the Structured Clinical Interview for the Psychotic Spectrum (SCI-PSY). The instrument is based on a spectrum model that emphasizes soft signs, low-grade symptoms, subthreshold syndromes, as well as temperamental and personality traits comprising the clinical and subsyndromal psychotic manifestations. The items of the interview include, in addition to a subset of the DSM-IV criteria for psychotic syndromes, a number of features derived from clinical experience and from a review of the phenomenological descriptions of psychoses. Study participants were enrolled at 11 Italian Departments of Psychiatry located at 9 sites and included 77 consecutive patients with schizophrenia or schizoaffective disorder, 66 with borderline personality disorder, 59 with psychotic mood disorders, 98 with non-psychotic mood disorders and 57 with panic disorder. A comparison group of 102 unselected controls was enrolled at the same sites. The SCI-PSY significantly discriminated subjects with any psychiatric diagnosis from controls and subjects with from those without psychotic disorders. The hypothesized structure of the instrument was confirmed empirically. PMID- 15885529 TI - Differential patterns of premorbid academic and social deterioration in patients with schizophrenia. AB - Schizophrenia is a neurodevelopmental disorder that is characterized by a number of behavioral abnormalities that are present prior to onset. These premorbid abnormalities may serve as unique markers for the disorder. The current study examines academic and social functioning prior to schizophrenia onset in a group of 58 males diagnosed with schizophrenia. The pattern of deterioration for social and academic functioning was examined across three age periods including childhood, early adolescence, and late adolescence, using the retrospective Premorbid Adjustment Scale (PAS). Results indicated that while increasing deterioration was present for both social and academic adjustment across age periods, there was a significant difference in deterioration between academic and social functioning (p<.05) during late adolescence, with greater deterioration in academic functioning. Results of the current study suggest that premorbid academic functioning is particularly susceptible to deterioration during late adolescence, with accelerating deterioration as schizophrenia onset becomes imminent. When considered together with results from other studies, the present findings suggest that deterioration in premorbid academic functioning from early to late adolescence may be a unique premorbid marker for schizophrenia. PMID- 15885530 TI - Dermatoglyphics and Schizophrenia: a meta-analysis and investigation of the impact of obstetric complications upon a-b ridge count. AB - BACKGROUND: Patients with schizophrenia show deviances in their dermatoglyphics, in particular reductions in palmar a-b ridge counts (ABRCs), which are evidence of an early developmental deviance. However, the severity or the origin of these ABRC changes has not been established. METHOD: (i) We examined the published literature on the ABRC in patients with schizophrenia against controls with a random effects meta-analysis. (ii) We used linear regression to study the ABRC in our sample of families including 125 patients with schizophrenia, 107 of their unaffected relatives and 98 controls. (iii) The effect of obstetric complications on the patient's ABRC was examined using the Lewis Murray scale. RESULTS: The pooled standardised effect size of ABRC differences between patients and controls obtained by our meta-analysis was 0.39 (95% CI: 0.05-0.73; p=0.03). In our sample, there were no significant differences in ABRCs between those with schizophrenia, their relatives and controls. Only those patients with obstetric complications had significantly reduced ABRC compared to controls (p=0.01). CONCLUSIONS: We confirmed the presence of significant yet mild ABRC reductions in schizophrenia. These represent a subtle deviance from the norm and could be present in certain subsets of patients, possibly those who suffered early developmental insults. PMID- 15885531 TI - The token economy for schizophrenia: review of the literature and recommendations for future research. AB - The token economy is a treatment intervention based on principles of operant conditioning and social learning. Developed in the 1950s and 1960s for long-stay hospital patients, the token economy has fallen out of favor since that time. The current review was undertaken as part of the 2003 update of the schizophrenia treatment recommendations of the Patient Outcomes Research Team (PORT). A total of 13 controlled studies of the token economy were reviewed. As a group, the studies provide evidence of the token economy's effectiveness in increasing the adaptive behaviors of patients with schizophrenia. Most of the studies are limited, however, by methodological shortcomings and by the historical context in which they were performed. More research is needed to determine the specific benefits of the token economy when administered in combination with contemporary psychosocial and psychopharmacological treatments. PMID- 15885532 TI - Socioeconomic status of origin and the clinical expression of Schizophrenia. AB - The concentrated focus on biology during the "decade of the brain" has resulted in decreased attention to the indisputable influence of psychosocial and sociocultural factors in the expression of schizophrenia. The aim of this study was to evaluate the relationship between the socioeconomic background of the origin and the clinical manifestation of schizophrenia. Parent socioeconomic information and clinical symptom data from 120 (84 men and 36 women) schizophrenia and schizoaffective patients were analyzed. Results suggested that higher parental SES (socioeconomic status) is associated with decreased symptom severity in female patients, but with increased symptom severity and decreased global functioning in male patients. Possible interactions between socioeconomic status and sex of patient are discussed, as well as suggestions for further study. PMID- 15885533 TI - Self-reported body weight perception and dieting practices in community-dwelling patients with schizophrenia. AB - INTRODUCTION: Many patients with schizophrenia are exposed to serious health risks associated with their excess body weight. Evidence exists that even a moderate amount of weight loss may have significant health benefits. Thus, weight control in schizophrenia patients has become an important treatment goal. Although studies in the general population show that satisfaction with body weight is an important predictor for engagement in various weight loss measures, the perspective of schizophrenia patients has not been assessed. METHOD: Information on self-reported weight perception, desire to lose weight as well as weight loss attempts was obtained according to methods employed in the National Health and Nutrition Examination Survey, Cycle III (NHANES III). Body weight and height were measured and body mass index (BMI) was calculated. RESULTS: Perception of body weight and desire to lose weight were correlated to BMI. Both obese female and male subjects (BMI30) were aware of their weight status. However, whereas overweight females (BMI>25< or =29.9) accurately perceived themselves so, males in this category had difficulties perceiving themselves overweight, and consequently neither wanted to lose weight, nor tried to lose weight. As means of weight loss, caloric restriction (diet) was most frequently employed (by more than 80% of study subjects); yet only a third of study subjects (34.4%) engaged in the recommended combination of diet and exercise to lose weight. Questionable weight loss practices were also frequently employed, especially among women. CONCLUSIONS: Obese patients (BMI> or =30) were generally aware of their excess body weight and wanted to lose weight. Only non-obese, yet overweight males (BMI>25< or =29.9) did not perceive themselves as overweight and consequently did not try to lose weight. Weight loss practices did not always follow established recommendations. Especially women were likely to approach weight loss with questionably appropriate and unsafe methods. PMID- 15885534 TI - Seasonality of schizophrenia births in the Japanese population: increased winter births possibly confined to the north area. AB - A number of North American and European studies have observed a higher proportion of winter births in schizophrenia patients. Thus, seasonal fluctuation of unknown environmental factors may affect brain development in ways that alter susceptibility to schizophrenia. Specification of these factors may help elucidate the etiopathological mechanism of the disease, about which little is certain. A small number of studies have investigated this issue in Asian populations, and the findings are not as consistent as those of Western populations. No remarkable excess of winter births has been observed in Japanese or Korean studies, while some studies have reported a significant decrease of summer births. We further investigated the issue in Japanese patients with schizophrenia (n=3927). No significant excess of winter births was observed, but a decrease in the summer births was found in male subjects. This is largely consistent with previous Japanese studies; however, when the subjects were confined to those born in a colder and higher latitude area of Japan (n=1338), a consistent trend for both a winter increase, and a summer decrease, was found. However, the results did not consistently reach statistical significance, possibly due to the lack of statistical power. Environmental factors that correlate with latitude might play a role in the development of the seasonality of births in schizophrenia. Further studies in a larger sample size are required to test these possibilities. PMID- 15885535 TI - Axis-I disorders and vulnerability to psychosis. AB - BACKGROUND: The psychopathology that manifests during the prodromal phase of first-episode psychosis is varied. Little is known about the clinical diagnoses of subjects with so-called prodromal or psychotic-like symptoms. METHOD: Samples of psychotic patients, first-degree relatives (FDRs) of psychotic, or severely ill patients, treatment-seeking patients, and a random community sample (in all 157 subjects) were assessed by the Structured Interview for Prodromal Symptoms (SIPS) and the SCID-I. Vulnerability to psychosis (VTP) was defined by severity of positive symptoms reported in the SIPS interview and associated with lifetime SCID-I diagnoses. RESULTS: The number of lifetime diagnoses received increased linearly as the SIPS symptoms approached more psychotic-like phenomena. All VTP subjects received on average 2.5, and currently prodromal subjects 2.9 lifetime SCID-I diagnoses, while the corresponding figure for non-VTP subjects was 0.7 (p<0.0001). Mood disorders and comorbid anxiety disorders were particularly common. CONCLUSION: Vulnerability to psychosis seems to be associated with a high number of lifetime Axis-I diagnoses. Occurrence of anxiety disorders is remarkable, and most VTP subjects can be diagnosed with a lifetime mood disorder. VTP subjects require careful assessment of mood and anxiety symptoms and adequate treatment for their multiple disorders. PMID- 15885536 TI - Seasonality of birth in schizophrenia patients in Poland. PMID- 15885537 TI - Symptomatic versus metabolic changes with pharmacotherapy of schizophrenia. PMID- 15885538 TI - [Advances in treatment of HIV infected children and adolescents. Barriers and bottlenecks]. PMID- 15885539 TI - [Genital bacterial carriage during the last trimester of pregnancy and early onset neonatal sepsis]. AB - Bacterial infections remain a major cause of morbidity and mortality in newborn infants. OBJECTIVE: To determine the bacterial ecology and pathological status of the genital organs during the last trimester of pregnancy and the germs of the following early-onset neonatal sepsis, in order to evaluate the risk of materno foetal infections and to find out a drug prophylaxis. METHOD: Vaginal and endocervical samples, usually taken during the first trimester of pregnancy were delayed and taken during the last trimester of pregnancy. A macroscopic examination described the aspect of the vagina, the cervix uteri, leukorrhea and of possible inflammatory lesions or ulcerations. A microscopic examination searched for parasites, epithelial cells, clue cells and leukocytes. The appropriate bacteriological cultures were performed after reading the Gram stain and scoring the vaginal flora. The clinical and cytobacteriological aspects were used to identify the bacterial ecology and the pathological genital states. An exploration was carried out in every newborn suspected of infection. RESULTS: Genital samples were collected from 306 pregnant women. Among them, 118 were at 29-32 weeks of gestation, 104 at 33-36, and 84 at 37-40. The most frequent germs were C. albicans (33,5%), Enterbacteriaceae (20.3%) including E. coli (10.9%), S. aureus (15.4%), Gardnerella (13.6%), and Trichomonas (10.6%), in monomicrobian (79.2%) and polymicrobian carriage (20.8%). Lower genital tract pathological states such as vaginitis (29.4%), bacterial vaginosis (21.5%) or endocervicitis (10.4%), asymptomatic bacterial carriage (23.5%) and normal genital flora (15%) were identified. These pregnancies led to 334 live births with 27 cases of early onset neonatal sepsis to which endocervicitis (25%) and vaginosis (19,7%) were most often linked. CONCLUSION: Genital samples at the last trimester of pregnancy could evaluate the risk of maternofoetal infections and allow to adapt a drug prophylaxis of Enterobacteriaceae, the most frequent germ of neonatal infections, as it has been done for Streptococcus agalactiae. But larger studies are required to evaluate the risk of maternofoetal infections and to state the drug prophylaxis. PMID- 15885540 TI - [Psychophysiological factors in high intellectual potential: comparative study in children aged from 8 to 11 years old]. AB - Factors of intellectual talent as well as physiological and psychological characteristics are little known. However, giftedness is now a social problem and the knowledge of precocity hallmarks should permit the diagnosis in order to undertake the adequate educational orientation of these children. From questionnaires given anonymously to parents, this work showed comparative results between a population of gifted children and a control one. OBJECTIVES: The aim of this study was to present precocity hallmarks in the gifted population, following factorial analysis of socio-economic and familial parameters, as well as medical and psycho-physiological variables linked with the children's post-natal life. POPULATION: Four hundred and twelve children aged eight to 11 years, from Lyon, have been used in the survey: 217 children (control group) were included after a random selection in schools, and 195 children (gifted group, IQ >130, Wechsler test) were enrolled after clinician's diagnoses in four medical departments. RESULTS: Regression analysis allowed to draw relation between several factors and giftedness: abnormal pregnancy (CR =3.205, P =0.009), perinatal stress (CR =2.166, P =0.003), and presence of migraine (CR =3.169, P =0.001). Parents living together (married or not) (CR =2.100, P =0.080) with a good and superior level of learning (CR =5.464, P =0.0002) were also linked with giftedness. CONCLUSION: Our results indicate that precocity hallmarks are multifactorial. These data confirm some socio-economic and medical physiological features correlated with giftedness, and suggest that psychological factors may bear on the etiology of intellectual talent. We focus on the hypothesis than early stress may play a role on central nervous system maturation in these children. PMID- 15885541 TI - [Infectious complications postengrafment in the first year after autologous stem cell transplantation in children]. AB - BACKGROUND: Studies on the infectious complications postengrafment in pediatric stem cell transplantation patients are rare. The aim of this study was to assess the incidence, types, outcome and factors affecting late infections. PATIENTS AND METHODS: A single-institution retrospective analysis was done of infections recorded in the first year following engrafment in children who underwent autologous stem cell transplantation for solid tumors from January 1991 to December 2000. A systematic antimicrobial chemoprophylaxis of TMP/SMX was administered. Patients who were at high risk for varicella-zona virus infection received prophylactic acyclovir. RESULTS: Eighty-four assessable patients were enrolled. Fifty-four patients (64%) underwent autologous peripheral blood stem cell transplantation and 30 patients (36%) underwent bone marrow transplantation. Forty-nine episodes of infections were documented in 39 patients (46%) of whom 27 patients (32%) developed infections after the first 100 days post transplantation. Bacterial septicemia occurred in nine patients of whom four patients had a catheter-related septicemia. Twelve patients (14%) developed localized herpes zoster infection. Local fungal infection occurred in five patients. Infection-related death occurred in one patient with non-documented pneumonitis. Univariable analysis showed a correlation between the CD34(+) cell dose <7.5 10(6)/kg and the development of infections (P =0.04). Patients who did not go into remission after transplantation where at high risk for septicemia (P =0.007). Multivariate analysis showed that a history of varicella or pretransplant varicella-zona positivity was the only significant factor for development zoster infection (P =0.01). CONCLUSION: Our study shows that infections in the first year postengrafment following autologous stem cell transplantation for solid tumors have a good prognosis and that the use of TMP/SMX should be the single systematic antimicrobial prophylaxis. The CD34(+) cell dose seems to play a role in preventing late infections. PMID- 15885542 TI - [Idiopathic hypothalamic syndrome: retrospective study and literature review]. AB - Hypothalamic obesity is usually induced by tumoral or genetic alterations such as craniopharyngioma or Prader-Willi syndrome, respectively. However, few cases have been reported without recognized etiology, this syndrome is also called idiopathic hypothalamic syndrome. OBJECTIVES: To improve definition and frequency of complications associated with this syndrome. POPULATION AND METHODS: A retrospective cohort study was performed in French endocrine paediatric departments and was associated with a literature review. RESULTS: We report five cases of idiopathic hypothalamic syndrome. This syndrome is correlated with a high mortality (one of our five cases, 25% in the literature) by neurovegetative dysfunction (breathing or thermal alteration). Obesity began before six years old because of compulsive eating and resulted in social behaviour disorders. Abnormal endocrine secretions were characterized by early hyperprolactinemia, permanent but later somatotrope deficiency and 80% of thyreotrope deficiency. Puberty abnormalities included hypogonadotropic hypogonadism as well as precocious (one of our cases, three cases including literature) or normal puberty. Neurogenic hypernatremia and water and electrolytic disorders were also responsible of acute neurological alterations. CONCLUSION: This largest study ever reported of idiopathic hypothalamic syndrome emphasizes the need of a multidisciplinary coordination to provide the best care of these patients. PMID- 15885543 TI - [Neonatal outcome after exposure to beta adrenergic blockers late in pregnancy]. AB - OBJECTIVES: To analyse neonatal effects after in utero betablockers exposure and the pertinence of recommendations delivered by our team. POPULATION AND METHODS: We report 44 pregnancies exposed to betablockers during the late pregnancy including the period of delivery about which the Regional Pharmacovigilance Center of Tours (CRPV) was questionned. RESULTS: Among the 39 children for whom we know the follow up, 22 had neonatal adverse effects of which 19 could be explained by in utero exposure to betablockers i.e. an hypoglycaemia (11 times), a bradycardia (six times), a bradycardia and hypoglycemia (one time) and an hypotension (one time). A drug-related effect was retained for eleven newborns (27%) and another etiology could be evoked in the eight others. The risk of neonatal adverse effects seems to increase in newborns exposed to labetalol (5/11), to betaxolol (1/2) or to propranolol (2/6) or when the dose is high. The eight newborns who had intrauterine growth retardation were generally more often exposed to atenolol than eutrophic newborns. Four babies had malformations. CONCLUSIONS: Our recommendation was an hospitalization 44 times (100%) to monitor heart rate, blood pressure and glycemia. When the follow-up is known, hospitalization was performed in 88% of the cases. Glycemia, heart rate and blood pressure were monitored in all the hospitalized children and in three of the five not hospitalized children. Our recommendation seems particularly justified with regard to hypoglycemia which is often asymptomatic but whose consequences can be severe. Atenolol often provider of intrauterine growth retardation and labetalol more often at the origin of neonatal adverse effects are probably to avoid during pregnancy. PMID- 15885544 TI - [Factors altering low-flow drug delivery using syringe pumps: Consequences on vasoactive drug infusion in preterm infant]. AB - The infusion of vasoactive drugs in preterm babies using syringe pumps can be complicated by important variations in the infusion rate. AIMS OF THE STUDY: 1) to assess the time between pump start and the actual presence of a substance in the circulation, when using slow rates; 2) to evaluate the effects of changes in the main infusion rate on substance rates infused on the same line. METHODS: We used 10% glucose to mimic vasoactive drugs. The main infusion consisted of saline, and both were infused on the same line using syringe pumps. Glucose concentration was measured at the end of the line. RESULTS: The time between pump start and the actual infusion of glucose depended on syringe volume and on the adjustment of the syringe in the pump. There was a transient increase in glucose rate when the main rate increased, and a transient decrease when the main rate decreased. CONCLUSION: To minimize the time before a substance is actually infused, one should use low volume syringes and correctly adjust the syringe in the pump. Moreover, when a steady infusion rate is necessary, one should avoid variations in the main infusion rate. PMID- 15885545 TI - [Neonatal diagnosis of primary ciliary dyskinesia: report of one case]. AB - Primary ciliary dyskinesia is a rare, genetic disorder resulting of an abnormal ultrastructural morphology of cilia. Such disease is rarely recognized in neonatal period. We report on a newborn who exhibited unexplained respiratory distress. The diagnosis of primary ciliary dyskinesia was suggested by the association of bilateral and multiple atelectasis and situs inversus. Diagnosis was confirmed by three months of age by ultrastructural study of cilia. Primary ciliary dyskinesia is a rare disease. Diagnosis should be considered in unexplained cases of neonatal respiratory distress, especially when situs inversus totalis and multiple atelectasis are present. Diagnosis requires ciliary studies that can be performed in newborn infants. PMID- 15885546 TI - [Traumatic perinatal cervical dislocation]. AB - We report on a case of cervical dislocation in which the severity of the lesion contrasts with the paucity of the symptoms. An eight-month-old girl presented with weakness of the left upper limb. She had a normal cephalic delivery. On the following days, increased weakness of the limb showed a spontaneous, partial regression. Clinical examination at eight months showed a normal physical and psychical development, a falling neck and an impossibility to maintain the sitting position, but a normal mobility of the lower limbs. Cervical radiography performed showed a complete dislocation. CT confirmed the diagnosis and eliminated congenital vertebral anomalies. Magnetic resonance imaging showed an important stretching and compression of the spinal cord. Cervical spine damage in the newborn are scarce and serious. The here reported case is characterized by a complete vertebral dislocation associated with medullar stretching and compression, but few initial symptoms. PMID- 15885547 TI - [Prenatal diagnosis of congenital mesoblastic nephroma. A case report]. AB - Antenatal ultrasounds allow the detection of renal tumors, especially renal mesoblastic nephromas, but only the pathological analysis of the surgical specimen can confirm this diagnosis postnatally. OBSERVATION: We report the prenatal discovery of a mesoblastic nephroma because of premature labour. Postnatal early surgery was decided because of possible complications in this premature infant. Histology revealed mesoblastic nephroma. COMMENTS: We point out the diagnostic elements of congenital mesoblastic nephroma, especially in what is related to arterial hypertension and hypercalcemia, histology and cytogenetics. PMID- 15885548 TI - [Langerhans cell histiocytosis associated with pituitary stalk transection]. AB - Clinical manifestations of hypothalamic-pituitary Langerhans'cell histiocytosis are commonly, diabetes insipidus and sometimes growth hormone deficiency. Their morphologic characteristics on magnetic resonance imaging are absence of posterior pituitary hyperintensity and thickening of the pituitary stalk. Pituitary stalk transection is characterized on magnetic resonance imaging by the absence of pituitary stalk visibility, hypoplasia of the anterior hypophysis and ectopic posterior pituitary hyperintense signal. This syndrome has been shown to be associated with either isolated growth hormone deficiency or multiple anterior pituitary hormone deficiency, but normal posterior pituitary function. CASE REPORT: We report our experience with a six-year-old boy who had been treated for three years for a multisystem Langerhans'cell histiocytosis with diabetes insipidus and who was admitted because of short stature. Endocrinological examinations demonstrated a profound growth hormone deficiency and a partial central hypocorticism. Magnetic resonance imaging showed pituitary stalk transection and a midline anomaly of the brain (Arnold Chiari type I malformation). CONCLUSION: Although, some events of his perinatal history lead to the hypothesis of a malformative origin, the progression of Langerhans' cell histiocytosis affected tissues to fibrosis, suggest that this disease is the cause of the patient's hypothalamohypophyseal lesions. PMID- 15885549 TI - [Cytochrome c oxydase-deficient Leigh syndrome with homozygous mutation in SURF1 gene]. AB - Leigh syndrome is a heterogeneous disorder, usually due to a defect in oxidative metabolism. Mutations in SURF1 gene have been identified in patients with cytochrome c oxidase deficiency. We report a homozygous splice site deletion [516 2_516-1delAG] in a young girl presenting with cytochrome c oxidase-deficient Leigh syndrome. Identification of molecular defect is indispensable for genetic counselling and prenatal diagnosis. PMID- 15885550 TI - [Lung protective strategies for premature infants]. AB - Despite many advances in perinatal medicine, bronchopulmonary dysplasia still frequently occurs in very premature infants. The very fragile lungs of these infants therefore have to be protected from birth. The protective strategies consist in applying positive expiratory pressure immediately, and using exogenous surfactant in a prophylactic or early use approach. The recent, variable flow, continuous positive airway pressure (CPAP) systems are very efficient and may allow to avoid tracheal intubation, or to facilitate weaning. When mechanical ventilation has to be used, high peak pressure and/or high tidal volume have to be avoided in order to prevent volutrauma. Accepting not to normalize PCO(2) contributes to it. High frequency oscillatory ventilation, which actually does not prevent bronchopulmonary dysplasia, is an extremely efficient ventilatory support technique for severe respiratory failure. Postnatal gluco-corticoid use reduces the rate of bronchopulmonary dysplasia at 36 weeks, but also results in an increased incidence of long-term neurological handicaps. In our experience, using these treatments can be avoided. Maternal transfer to a level three perinatal center, associated with the adequate use of theses lung protective strategies following very premature birth enable the less unfavorable results to be obtained. PMID- 15885551 TI - [Gustatory perceptions in children]. AB - As soon as the end of gestation, the gustatory system is stimulated by the taste active compounds carried by the amniotic fluid and its maturation continues until mid-childhood. Facial expressions and relative ingestion methods show that the newborn can discriminate the various taste qualities (bitter, salty, sour, sweet and umami). The range of individual responses is wide. Neonatal reactions to sweet and umami are generally considered to express pleasure. The bitter and sour stimulations lead to hedonically negative reactions. The response to salt taste is less characteristic. Overall, the attraction towards sweet and the rejection of bitter and sour tastes become more pronounced during childhood but tend to decrease in adult life. The early attraction to sweetness is reinforced by exposure to sweet stimulations. With age, the response to salt evolves towards attraction which intensity is dependent on the context and on postnatal exposures to salt. The link between gustatory sensitivity to sweet, salty and sour stimuli and food preferences is far from being clear; the sensitivity to bitter taste better explains the rejection of bitter foods, such as vegetables for instance. The development of gustatory perceptions partly depends upon experience. A better knowledge of the role of experience could help to improve the orientation and the efficacy of nutritionally-oriented food education strategies. PMID- 15885552 TI - [Diagnosis and treatment of juvenile osteoporosis]. AB - Bone mass is primarily genetically determined, but exogenous factors also play a major role. The prevention of osteoporosis can start from childhood, and optimal achievement of peak bone mass during childhood and adolescence is important in order to minimise future fracture risks. Chronic inflammatory diseases can have a detrimental effect on bone mass, by means of several mechanisms. Different diagnostic methods for detection and monitoring of osteoporosis are in use or under investigation. The role of calcium and vitamin D supplementation for the prevention and treatment of osteoporosis associated with paediatric rheumatic diseases remains to be established. New treatments such as bisphosphonates and calcitonin are now available, although their use in the paediatric age has been limited. PMID- 15885553 TI - [Management of phenylketonuria and hyperphenylalaninemia: the French guidelines]. AB - Phenylketonuria (PKU) is an inherited metabolic disease affecting about one birth out of 15 000. From 1978, a national systematic neonatal screening was set up in France with a regional organisation. French rational and guidelines have been established by the national PKU group with the collaboration of all the physicians responsible for the regional centres. These guidelines specify the minimal diagnosis procedures leading to an optimal treatment of all patients. A low-phenylalanine diet must be started as soon as possible in the neonatal period for all newborns whose phenylalanine levels are above 10 mg/dl. The dietary control must keep the phenylalanine plasma levels between 2 and 5 mg/dl until 10 years of age. After this age, several data argue for a progressive and controlled relaxation of the diet, keeping the phenylalanine level below 15 mg/dl until the end of the adolescence and below 20 to 25 mg/dl in adulthood. All PKU patients must be followed up for life, in order to screen those who may not bear the diet relaxation and in order to strictly prevent maternal PKU deleterious consequences. PMID- 15885554 TI - [Radiological quiz of the month. A sudden abdominal distension in a four-year-old child]. PMID- 15885555 TI - [Professional activity of fellows in French pediatric intensive care units]. PMID- 15885556 TI - [Group B coxsackie virus infection with fatal myocarditis in the neonate after early discharge from maternity unit]. PMID- 15885557 TI - [Skin tests in allergic children of Ivory Coast]. PMID- 15885558 TI - [Rational use of viral serology in the child]. AB - Very sensitive and allowing discrimination between IgG and IgM, the enzyme immunoassays are the most commonly used to carry out viral serologies. The presence of IgG corresponding to a natural or postvaccination immune status or to a latent or occult infection is a reliable information to assess the protection against some opportunistic or teratogenic infections respectively in children or pregnant women. The search of IgM by immunocapture is suitable to early diagnose the majority of common viral diseases in childhood, specially when presenting in an atypical form, and to attest a congenital rubella infection. Moreover, IgG avidity proved to be useful in dating an infection and confirming positive IgM results. However, viral infections localized at an entry site of the host, such as respiratory and enteric diseases, as well as syndromes of herpes simplex and enteroviral origin, do not represent indications of serological diagnosis. Provided its use with discernment, the serological diagnosis of viral infections have to be considered as a high performance and of low cost biological tool. PMID- 15885559 TI - [Migraine and headache in childhood]. AB - In childhood and adolescence, migraine is the main primary headache. This diagnosis is extensively underestimated and misdiagnosed in pediatric population. Lacks of specific biologic marker, specific investigation or brain imaging reduce these clinical entities too often to a psychological illness. Migraine is a severe headache evolving by stereotyped crises associated with marked digestive symptoms (nausea and vomiting); throbbing pain, sensitivity to sound, light are usual symptoms; the attack is sometimes preceded by a visual or sensory aura. During attacks, pain intensity is severe, most of children must lie down. Abdominal pain is frequently associated, rest brings relief and sleep ends often the attack. The prevalence of the migraine varies between 5% and 10% in childhood. At childhood, headache duration is quite often shorter than in adult population, it is more often frontal, bilateral (2/3 of cases) that one-sided. Migraine is a disabling illness: children with migraine lost more school days in a school year, than a matched control group. Migraine episodes are frequently triggered by several factors: emotional stress (school pressure, vexation, excitement: upset), hypoglycemia, lack of sleep or excess (week end migraine), sensorial stimulation (loud noise, bright light, strong odor, heat or cold...), sympathetic stimulation (sport, physical exercise). Attack treatments must be given at the early beginning of the crisis; oral dose of ibuprofen (10 mg/kg) is recommended. If the oral route is not available when nausea or vomiting occurs, the rectal or nasal routes have then to be used. Non pharmacological treatments (relaxation training, self hypnosis, biofeedback) have shown to have good efficacy as prophylactic measure. Daily prophylactic pharmacological treatments are prescribed in second line after failure of non-pharmacological treatment. PMID- 15885560 TI - [Interest of EEG in full-term newborns with isolated unilateral ischemic stroke]. AB - In full-term newborns, unilateral and recurrent clonic seizures which occur during the first 48 hours of life suggest an isolated unilateral ischemic stroke. These focal seizures are isolated, occuring during a short period, or make up a status epilepticus. Electroencephalogram (EEG) is a key tool for crisis and focal cerebral process diagnosis. It also allows to assess antiepileptic drugs effectiveness. But cerebral imaging is necessary to confirm vascular origin of the cerebral impairment. Hematologic data are also needful to look for a family thromphilia. Some particular unilateral EEG abnormalities could be associated with controlateral motor sequelae or long term behavioral problems. These findings may be used for prospective studies aimed at specifying possible links between EEG abnormalities and long term outcome. PMID- 15885561 TI - [Contribution of day care general anaesthesia for dental care in children and handicapped patients]. AB - Dental surgeons, pediatrists and doctors daily note the precocity and the importance of caries in children. This evolution makes essential the general anaesthesia in paediatric dentistry among the therapeutic arsenal of pedodontists. Fortunately, with the greatest knowledge of the child physiology, the better control of the anaesthetic operative protocols, general anaesthesia is very efficient. Furthermore, the development of the day care treatment now allows the access of these treatments to an increasingly large public. For pedodontists and anaesthetist, generals anaesthesia is essentially indicated for children with many decays, presenting behaviour disorders, a physical or cerebral handicap, a turmoil engraves general state. This operating mode aims to restore: This particular aspect of the dental practice remains still too little developed and cannot answer the demands. The necessity made it a rule to set up a theoretical and practical training. This specialized dentistry waits for recognition on behalf of supervisory organisms and a similar message spread by all nursing persons. PMID- 15885563 TI - A framework for distributed mediation of temporal-abstraction queries to clinical databases. AB - OBJECTIVE: The specification and creation of a distributed system that integrates medical knowledge bases with time-oriented clinical databases; the goal is to answer complex temporal queries regarding both raw data and its abstractions, such as are often required in medical applications. METHODS: (1) Specification, design, and implementation of a generalized access method to a set of heterogeneous clinical data sources, by using a virtual medical-record interface and by mapping the local terms to a set of standardized medical vocabularies; (2) specification of a generalized interface to a set of knowledge sources; (3) specification and implementation of a service, called ALMA that computes complex time-oriented medical queries that include both raw data and abstractions derivable from it; (4) design and implementation of a mediator, called IDAN, that answers raw-data and abstract queries by integrating the appropriate clinical data with the relevant medical knowledge and uses the computation service to answer the queries; (5) an expressive language that enables definition of time dependent medical queries, which are referred to the mediator; (6) evaluation of the effect of the system, when combined with a new visual interface, called KNAVE II, on the speed and accuracy of answering a set of complex queries in an oncology sub domain, by a group of clinicians, compared to answering these queries using paper or an electronic spreadsheet. RESULTS: We have implemented the full IDAN architecture. The IDAN/KNAVE-II combination significantly increased the accuracy and speed of answering complex queries about both the data and their abstractions, compared to the standard tools. CONCLUSION: The implemented architecture proves the feasibility of the distributed integration of medical knowledge sources with clinical data of heterogeneous sources. The results suggest that the proposed IDAN modular architecture has potential significance for supporting the automation of clinical tasks such as diagnosis, monitoring, therapy, and quality assessment. PMID- 15885564 TI - Temporal data mining for the quality assessment of hemodialysis services. AB - OBJECTIVE: This paper describes the temporal data mining aspects of a research project that deals with the definition of methods and tools for the assessment of the clinical performance of hemodialysis (HD) services, on the basis of the time series automatically collected during hemodialysis sessions. METHODS: Intelligent data analysis and temporal data mining techniques are applied to gain insight and to discover knowledge on the causes of unsatisfactory clinical results. In particular, two new methods for association rule discovery and temporal rule discovery are applied to the time series. Such methods exploit several pre processing techniques, comprising data reduction, multi-scale filtering and temporal abstractions. RESULTS: We have analyzed the data of more than 5800 dialysis sessions coming from 43 different patients monitored for 19 months. The qualitative rules associating the outcome parameters and the measured variables were examined by the domain experts, which were able to distinguish between rules confirming available background knowledge and unexpected but plausible rules. CONCLUSION: The new methods proposed in the paper are suitable tools for knowledge discovery in clinical time series. Their use in the context of an auditing system for dialysis management helped clinicians to improve their understanding of the patients' behavior. PMID- 15885565 TI - Generalised reliability characteristics for probabilistic networks. AB - BACKGROUND: In the medical domain, establishing a diagnosis typically amounts to reasoning about the unobservable truth, based upon a set of indirect observations from diagnostic tests. A diagnostic test may not be perfectly reliable, however. To avoid misdiagnosis, therefore, the reliability characteristics of the test should be taken into account upon reasoning. OBJECTIVE: In this paper, we address the issue of modelling the reliability characteristics of diagnostic tests in a probabilistic network. METHOD: To this end, we study the mathematical foundation of a test's characteristics and collate them with the probabilities required for a probabilistic network. RESULTS: We show that the standard reliability characteristics that are generally available from the literature have to be further detailed and stratified, for example by experts, before they can be included in a network. We demonstrate these modelling issues by means of a real life probabilistic network in oncology. PMID- 15885566 TI - Decision support of inspired oxygen selection based on Bayesian learning of pulmonary gas exchange parameters. AB - OBJECTIVE: To investigate if the real-time Bayesian learning of physiological model parameters can be used to support and improve the selection of inspired oxygen fraction. METHODS AND MATERIAL: Supporting the selection of inspired oxygen fraction relies on predictions of arterial oxygen saturation. The efficacy of using these predictions to select inspired oxygen was tested retrospectively in a system for estimating gas exchange parameters of the lung (Automatic Lung Parameter Estimator, ALPE). For the predictions to offer effective decision support they need to be accurate and above all safe. These qualities were tested with data from 16 post-operative cardiac patients, using two different tests. The aim of the first test was to assess retrospectively if the predictions could have supported clinical decisions. The second test sought to establish if the predictions could support improving the efficiency of inspired oxygen selection during an ALPE oxygen titration. RESULTS: The predictions were found to be reasonably accurate, and most importantly safe in both of the tests. CONCLUSION: The method described can be used to support the selection of inspired oxygen fraction, and it has the potential to improve the efficiency of inspired oxygen selection during an oxygen titration. PMID- 15885567 TI - A new method for sleep apnea classification using wavelets and feedforward neural networks. AB - OBJECTIVES: This paper presents a novel approach for sleep apnea classification. The goal is to classify each apnea in one of three basic types: obstructive, central and mixed. MATERIALS AND METHODS: Three different supervised learning methods using a neural network were tested. The inputs of the neural network are the first level-5-detail coefficients obtained from a discrete wavelet transformation of the samples (previously detected as apnea) in the thoracic effort signal. In order to train and test the systems, 120 events from six different patients were used. The true error rate was estimated using a 10-fold cross validation. The results presented in this work were averaged over 100 different simulations and a multiple comparison procedure was used for model selection. RESULTS: The method finally selected is based on a feedforward neural network trained using the Bayesian framework and a cross-entropy error function. The mean classification accuracy, obtained over the test set was 83.78+/-1.90%. CONCLUSION: The proposed classifier surpasses, up to the author's knowledge, other previous results. Finally, a scheme to maintain and improve this system during its clinical use is also proposed. PMID- 15885568 TI - Constraint reasoning in deep biomedical models. AB - OBJECTIVE: Deep biomedical models are often expressed by means of differential equations. Despite their expressive power, they are difficult to reason about and make decisions, given their non-linearity and the important effects that the uncertainty on data may cause. The objective of this work is to propose a constraint reasoning framework to support safe decisions based on deep biomedical models. METHOD: The methods used in our approach include the generic constraint propagation techniques for reducing the bounds of uncertainty of the numerical variables complemented with new constraint reasoning techniques that we developed to handle differential equations. RESULTS: The results of our approach are illustrated in biomedical models for the diagnosis of diabetes, tuning of drug design and epidemiology where it was a valuable decision-supporting tool notwithstanding the uncertainty on data. CONCLUSION: The main conclusion that follows from the results is that, in biomedical decision support, constraint reasoning may be a worthwhile alternative to traditional simulation methods, especially when safe decisions are required. PMID- 15885569 TI - Protein kinase C isozymes in stroke. AB - Stroke is a devastating neurologic disease and a leading cause of death and disability worldwide. Thrombolytic agents have been used to re-establish circulation in thromboembolic stroke, but their utility is limited by hemorrhage and reperfusion injury. Studies with experimental stroke models, mouse genetics, and selective peptide inhibitors and activators have implicated protein kinase C (PKC) epsilon in ischemic preconditioning and PKCdelta and gamma in tissue injury. PKCdelta, resident both in neutrophils and in the brain, appears particularly essential for reperfusion injury, and recent work using PKCdelta specific peptide inhibitors suggests that PKCdelta inhibitors could prove useful in attenuating reperfusion injury and improving outcome following thrombolysis. PMID- 15885570 TI - Molecular inroads into the anterior heart field. AB - In 2001, three research groups described a previously unrecognized population of progenitor cells in pharyngeal mesoderm that gives rise to myocardium at the arterial pole of the heart. In the last 4 years, the major importance of the cellular contribution of pharyngeal mesoderm to normal and pathologic heart development has become apparent. Lineage-tracing experiments have defined the extent to which pharyngeal progenitor cells colonize the heart, revealing a contribution to venous, as well as arterial, pole myocardium; in addition, major molecular inroads have been made into understanding gene regulation in pharyngeal myocardial progenitor cells, implicating forkhead, Gata, LIM homeodomain, MEF2, SMAD, and T-box transcription factors. The key role of the anterior heart field during normal heart development is underscored by the demonstration that both direct and indirect perturbation of myocardial progenitor cells in pharyngeal mesoderm result in congenital heart disease. PMID- 15885571 TI - Homing to hypoxia: HIF-1 as a mediator of progenitor cell recruitment to injured tissue. AB - The identification of bone marrow-derived endothelial progenitor cells has altered our understanding of new blood vessel growth and tissue regeneration. Previously, new blood vessel growth in the adult was thought to only occur through angiogenesis, the sprouting of new vessels from existing structures. However, it has become clear that circulating bone marrow-derived cells can form new blood vessels through a process of postnatal vasculogenesis, with endothelial progenitor cells selectively recruited to injured or ischemic tissue. How this process occurs has remained unclear. One common element in the different environments where vasculogenesis is believed to occur is the presence of a hypoxic stimulus. We have identified the chemokine stromal cell-derived factor-1 (SDF-1) and its receptor CXCR4 as critical mediators for the ischemia-specific recruitment of circulating progenitor cells. We have found that the endothelial expression of SDF-1 acts as a signal indicating the presence of tissue ischemia, and that its expression is directly regulated by hypoxia-inducible factor-1. Stromal cell-derived factor 1 is the only chemokine family member known to be regulated in this manner. Later events, including proliferation, patterning, and assembly of recruited progenitors into functional blood vessels, are also influenced by tissue oxygen tension and hypoxia. Interestingly, both SDF-1 and hypoxia are present in the bone marrow niche, suggesting that hypoxia may be a fundamental requirement for progenitor cell trafficking and function. As such, ischemic tissue may represent a conditional stem cell niche, with recruitment and retention of circulating progenitors regulated by hypoxia through differential expression of SDF-1. PMID- 15885572 TI - Adventitial progenitor cells contribute to arteriosclerosis. AB - Accumulating evidence indicates the involvement of vascular progenitor cells in the development of arteriosclerosis, including transplant arteriosclerosis, angioplasty-induced restenosis, vein graft atherosclerosis, and spontaneous atherosclerosis. Recently, it was found that the adventitia of the arterial wall contains a large number of progenitor cells, which can differentiate into smooth muscle cells in vitro and in vivo. These progenitor cells were able to migrate from the adventitia into the intima, where they accumulate to contribute to atherosclerotic lesions of vein grafts in apoE-deficient mice. Thus, these cells may be a source of smooth muscle cells and might have implications for cellular, genetic, and tissue engineering approaches to vascular disease. PMID- 15885573 TI - The reperfusion injury salvage kinase pathway: a common target for both ischemic preconditioning and postconditioning. AB - Novel treatment approaches, as potential adjunctive therapy to current reperfusion strategies (such as thrombolysis, primary coronary angioplasty, and cardiac surgery), are required to provide further cardioprotection in the setting of an acute myocardial infarction to effect further reductions in morbidity and mortality. In this regard, the activation of prosurvival kinases, such as Akt and Erk1/2 (which we have termed the reperfusion injury salvage kinase [RISK] pathway), at the time of reperfusion, has been demonstrated to confer powerful cardioprotection against myocardial ischemia-reperfusion injury. In this review, we present evidence suggesting that the cardioprotective phenomena of ischemic preconditioning and the recently described ischemic postconditioning exert their cardioprotective effects through the recruitment of the RISK pathway, at the time of reperfusion, and that the protection in these settings is mediated through the inhibition of mitochondrial permeability transition pore (mPTP) opening at this time. Therefore, the pharmacologic manipulation of the RISK pathway at the time of reperfusion may enable one to harness the powerful cardioprotective benefits of both ischemic preconditioning and postconditioning, and provide a novel approach to cardioprotection. PMID- 15885574 TI - Regulation of hematopoietic stem cells by the niche. AB - The quiescent state in the cell cycle is thought to be indispensable for the maintenance of hematopoietic stem cells (HSCs). Interaction of HSCs with their particular microenvironments, known as niches, is critical for maintaining the stem cell properties of HSCs, including cell adhesion, survival, and cell division. Hematopoietic stem cells balance quiescence and cell division in the stem cell niche and also maintain the potential for long-term hematopoiesis. We have recently reported that HSCs expressing the receptor tyrosine kinase Tie2 are in the G0 phase and anti-apoptotic, and comprise a side-population (SP) of HSCs, which contacts osteoblasts (OBs), the source of the angiopoietin-1 (Ang-1) ligand for Tie2 in the bone marrow (BM) niche. Tie2/Ang-1 signaling occurs in interactions between HSCs and niche cells. The interaction of Tie2 with Ang-1 in vitro induces tight adhesion of HSCs to stromal cells and is sufficient to maintain the long-term blood-repopulating (LTR) activity of HSCs in vivo by preventing cell division. In addition, Ang-1 enhances the ability of HSCs to become quiescent and induces their adhesion to the bone surface in vivo, resulting in protection of the HSC compartment from stresses suppressing hematopoiesis. These data suggest that the Tie2/Ang-1 signaling pathway plays a critical role in the maintenance of HSCs in the adult BM niche. Ang-1 produced by OBs activates Tie2 on HSCs and promotes tight adhesion of HSCs to the niche, resulting in quiescence and enhanced survival of HSCs. PMID- 15885575 TI - Oral estramustine plus oral etoposide in the treatment of hormone refractory prostate cancer patients: a phase II study with a 5-year follow-up. AB - BACKGROUND: Chemotherapy regimens that target microtubular trafficking were repeatedly found to be active in the treatment of hormone refractory prostate cancer patients, but disease responses were reportedly short-lived on average. MATERIALS AND METHODS: From 1994 to 1997, 46 consecutive patients with hormone refractory prostate cancer were enrolled in a multicenter Phase II trial of oral etoposide 100 mg/day and estramustine 560 mg/day for 21 days, followed by a 7-day rest period. Final evaluation of this trial was performed after a follow-up of 5 years. RESULTS: Fifty-four percent of patients attained a PSA response and 46% attained a response on measurable lesions. Median time to progression (TTP) and overall survival were 7.4 and 18.4 months, respectively. Fourteen patients (30.4%) had a TTP greater than 12 months and 9 (19.5%) a TTP greater than 18 months. Sixteen patients (34.8.%) survived more than 2 years and 2 (4.3%) survived more than 5 years. One patient was still alive and free from progression more than 7 years after starting treatment. CONCLUSIONS: This Phase II trial with a long-term follow-up revealed that some patients with hormone refractory prostate cancer could obtain durable disease response and long survival with an oral etoposide and estramustine combination regimen. PMID- 15885576 TI - PSA elevation during prostate cryosurgery and subsequent decline. AB - PURPOSE: To determine the immediate effect of prostate cryosurgery on PSA and the subsequent decline. METHODS AND MATERIALS: PSA level was measured in 14 patients who underwent cryosurgery for prostate cancer. Blood samples were taken immediately before and after cryosurgery and 1, 2, 4, 6 weeks and 3 months postoperatively. A confidence interval for the elevation from baseline to maximal PSA was calculated. PSA decline was assessed in patients with a nadir PSA < or =0.5 ng/mL. Patients with a postoperative nadir PSA >0.5 ng/mL were considered to harbor viable cancer and were excluded from the decline analysis. The observed PSA levels during the postoperative period were compared with the expected levels that were calculated according to the maximal PSA level and a serum half-life of 2.5 days. Student t-test was used to compare expected and observed PSA levels. RESULTS: PSA increased from an average of 9.23 ng/mL preoperatively to a maximum average of 155 ng/mL (maximal PSA ranges: 18.9-490.5 ng/mL). The 95% CI for the increase in PSA from baseline level was 63.4 to 224.14. PSA nadir < or =0.5 ng/mL was achieved in 10 patients. The observed PSA decline course was slower than expected according to its half-life. Average observed and expected PSA levels at 2,4 and 6 weeks after cryosurgery were 10.4 versus 3.57 ng/mL (P = 0.005), 0.65 versus 0.07 (P = 0.007) and 0.09 versus 0.001 (P = 0.03), respectively. CONCLUSION: PSA levels increase steeply following cryosurgery, and decline slower than expected according to the serum half-life. PMID- 15885577 TI - Long-term complications associated with the Indiana pouch urinary diversion in patients with recurrent gynecologic cancers after high-dose radiation. AB - Few studies have assessed the long-term risks associated with the Indiana pouch continent urinary diversion after high-dose radiation therapy. A retrospective review of consecutive female patients who underwent cystectomy and Indiana pouch urinary diversion identified 12 with a history of high-dose pelvic irradiation (mean total 78.1 Gy). Long-term complications and outcomes in this group were compared to a synchronous group of patients (n = 14) with no history of radiation. Mean follow-up in the radiation therapy (RT) and nonirradiated comparison group (CG) were 48.5 and 40.8 months, respectively, with all patients having over 12 months of outcomes assessed. In the RT group, 83% of patients experienced a one or more complications (n = 29) while 57% of the CG did (n = 15; P = 0.2). Complications seen more commonly in the RT group included ureteral stricture/obstruction (5 vs. 2), renal insufficiency (3 vs. 1) and severe incontinence (3 vs. 0). Notably, 23 secondary operative procedures were required in the RT group versus CG (n = 11, P = 0.2). Percutaneous nephrostomy (6 vs. 1; P = 0.03) and ureteral reimplantation (4 vs. 0; P = 0.03) were seen significantly more commonly in the RT group than the CG. Long-term follow-up is critical to assess the complications associated with urinary diversions. We conclude that frequent complications and a significant increase in specific operative procedures are observed in heavily irradiated patients with recurrent gynecologic cancers receiving an Indiana Pouch urinary diversion. Given the risk of renal insufficiency, close monitoring of renal drainage and function is recommended. These considerable long-term complications should be considered when counseling patients contemplating Indiana Pouch urinary diversion after radiation therapy. PMID- 15885578 TI - Evaluation of precursor prostate-specific antigen isoform ratios in the detection of prostate cancer. AB - OBJECTIVE: Disease-associated isoforms of the prostate-specific antigen (PSA) have recently been identified. We evaluated the efficacy of using precursor isoforms of PSA (pPSA) and their ratios for the detection of prostate cancer. METHODS: Serum concentrations of [-2], [-4], and [-7]pPSA, BPSA, and free PSA (fPSA) were retrospectively measured in 43 selected men. Of the 43 men, 15 had clinical T2 prostate cancer with ultrasound-estimated prostate volumes (PVs) of >50 cm(3), 13 had clinical T2 prostate cancer with (PVs) <25 cm(3), and 15 were prostate cancer-free with PV >50 cm(3). We calculated sum pPSA ([-2]+[-4]+[ 7]pPSA). We also compared the ratios of: free/total PSA, [-2]pPSA/fPSA, [ 2]pPSA/BPSA, [-2]pPSA/(fPSA-BPSA), [-2]pPSA/(fPSA-sum pPSA), and [-2]pPSA/{fPSA (sum pPSA+BPSA)} among these three groups. RESULTS: The median [-2]pPSA/(fPSA-sum pPSA) ratio was significantly higher in men with prostate cancer with or without large PV compared with men with large PV without prostate cancer. Values for median [-2]pPSA/free PSA ratio were higher in men with prostate cancer with or without large PV compared with men with large PV, and without prostate cancer, but the differences were not statistically significant. CONCLUSIONS: In this preliminary study, [-2]pPSA/(fPSA-sum pPSA) ratio was not associated with prostate gland volume but was associated with prostate cancer. This ratio may be useful in the detection of prostate cancer, particularly in men with larger glands. PMID- 15885579 TI - Leiomyosarcoma of the renal vein: case report and review of the literature. AB - Leiomyosarcomas of vascular origin is a rare and aggressive pathology, its presentation on the renal vein is infrequent, of which 29 cases have been reported world-wide. We describe a case of a 76 year-old woman, who presented with intermittent left flank pain, irradiated to the lumbar region of 6 months duration. A left radical nephrectomy with en-bloc resection of a para-aortic tumor was performed. Microscopic examination of the specimen reported a moderately differentiated leiomyosarcoma, originated in the wall of the renal vein. The patient received adjuvant chemotherapy and radiation therapy. The tumor recurred in the liver. After 24 months from the primary surgery, the patient is alive with evidence of multiple metastatic spread to the liver. She is now under palliative care. PMID- 15885581 TI - An introduction to dietary/supplemental omega-3 fatty acids for general health and prevention: part I. AB - The correction of a subtle nutritional deficiency that may reduce the risk of a future chronic disease is indeed a challenge. However, some specific examples in the past, such as the addition of folic acid to prevent neural tube defects and calcium and vitamin D to prevent osteoporosis, should provide some encouragement that some conditions can be prevented with the appropriate addition of a deficient compound. One of the most intriguing current and future impacts on public health may come from a greater intake of omega-3 fatty acids such as alpha linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). The omega-3 fatty acids continue to accumulate research that suggests that may prevent a variety of diverse chronic diseases and potentially some acute clinical scenarios. In Part 1 of this manuscript the potential for these compounds to prevent certain cardiovascular conditions are discussed. In Part 2 the potential for an impact in arthritis, numerous areas of cancer research, depression, maternal and child health, neurological diseases, osteoporosis, and other medical disciplines are also briefly covered. The future appears bright for these agents, but specifically which conditions, who qualifies, testing, frequency, adequate sources, future trials and numerous other questions need to be addressed and answered before the potential impact can catch up to the recent hype. PMID- 15885582 TI - An introduction to dietary/supplemental omega-3 fatty acids for general health and prevention: part II. AB - The correction of a subtle nutritional deficiency that may reduce the risk of a future chronic disease is indeed a challenge. However, some specific examples in the past, such as the addition of folic acid to prevent neural tube defects and calcium and vitamin D to prevent osteoporosis, should provide some encouragement that some conditions can be prevented with the appropriate addition of a deficient compound. One of the most intriguing current and future impacts on public health may come from a higher intake of omega-3 fatty acids, such as alpha linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). The omega-3 fatty acids continue to accumulate research that suggests that they may prevent a variety of diverse chronic diseases and potentially some acute clinical scenarios. In the first part of this article, the potential for these compounds to prevent certain cardiovascular conditions are discussed. In the second part, the potential for an impact in arthritis, numerous areas of cancer research, depression, maternal and child health, neurologic diseases, osteoporosis, and other medical disciplines are also briefly covered. The future appears bright for these agents, but specifically which conditions, who qualifies, testing, frequency, adequate sources, future trials, and numerous other questions need to be addressed and answered before the potential impact can catch up to the recent hype. PMID- 15885583 TI - Statins and cholesterol lowering after a cancer diagnosis: why not? AB - The pleiotropic effects of statins continue to garner attention. One area of interest that requires more attention is their potential, or that of another heart healthy agent, to impact favorably or inhibit the progression of numerous cancers, especially prostate cancer. Statins have accumulated a plethora of data in the area of general cancer cell line inhibition, and the potential to have a synergistic impact in patients receiving a variety of conventional treatments for a diversity of cancers is endorsed at least by laboratory and some preliminary small clinical studies. However, prostate cancer is unique compared to other cancers because, despite a high prevalence, some men choose watchful waiting or no treatment, especially if they are older and/or have a well-differentiated tumor. Thus, prostate cancer is a good candidate for the potential investigation of statins after diagnosis, with or without standard therapy. Regardless, because the number 1 or 2 cause of death in men with prostate cancer is cardiovascular disease than even in the worst-case scenario, these agents appear attractive for more immediate clinical study because they could impact a major cause of morbidity and mortality in these men. It is time to see the forest over the tree, and statins seem to have an argument as good as many other agents for allocating more money and time to test their ability as adjuvant therapy in a randomized trial or, in some cases, of nonaggressive disease as a potential monotherapy. PMID- 15885584 TI - Promoting general health during androgen deprivation therapy (ADT): a rapid 10 step review for your patients. AB - Androgen deprivation for prostate cancer use to be applied only in the latter stage of the disease process, thus, the issue of promoting general health during this time was not a concern because the subject of life and death was more paramount. However, thanks to earlier detection of prostate cancer, there has been a general stage migration in this disease. Men are choosing these traditionally late stage therapies earlier and earlier. Therefore, the subject of quality of life on this treatment has now garnered as much attention as the survival issues. Cognitive or mental health concerns, cholesterol changes, hot flashes, osteoporosis, and other side effects are being addressed and treated with a variety of conventional medicines. However, the issue of the role of the patient or what men can do personally to promote better mental and physical health is desperately needed in this area. A variety of beneficial lifestyle changes and over-the-counter agents may have an enormous impact on men's health during androgen deprivation. Calcium and vitamin D supplements, aerobic and resistance exercise, cholesterol awareness and reduction, weight loss, and other individual changes could have an enormous impact on the quality and quantity of a man's life. Some of these so called "bottom line" recommendations are reviewed in this article to empower the patient during this time, and to send clearly the message that he has a role to play apart from just picking up and using a prescription drug for side effects, and his role is just as critical for improving the probability of living longer and better. PMID- 15885585 TI - New approaches in advanced renal cell carcinoma. PMID- 15885601 TI - Incidence of malignancies among patients with type I Gaucher disease from a single referral clinic. AB - BACKGROUND: It has been implied that the incidence of malignant disorders is increased in patients with non-neuronopathic (type I) Gaucher disease. The purpose of the study was to ascertain numbers of cancers in type I Gaucher disease since this is of considerable concern to patients and physicians. METHODS: Records of 505 patients with type I Gaucher disease seen at a large referral clinic since 1990 were culled in December 2004 to ascertain diagnosis of a cancer identified during follow-up. Age-matched data from the Israeli National Cancer Registry (INCR) database were used for comparison. FINDINGS: Patients diagnosed with cancer before 1990 were not included. Of the remaining 500 patients, 227 (45.4%) were male, mean age=38.7 years; and 273 (54.6%) were female, mean age=37.0 years (SD=21.0 years for both). Twenty patients (4.0%) had developed a cancer through December 31, 2003: 6 were male and 14 were female. The most common were three cases each of lymphoma and myelodysplastic syndrome and two cases of multiple myeloma. There was no statistically significant excess of cancer rate among patients relative to age-matched rates reported in national Jewish Israeli and Ashkenazi Jewish Israeli registry records. CONCLUSIONS: There appears to be no excess risk for hematological or other cancers among patients with type I Gaucher disease relative to the overall Jewish population matched for age. This study confirms recent international studies of patients with Gaucher disease for no excess risk for all cancers but multiple myeloma where these latter studies implicate a significantly higher incidence. PMID- 15885602 TI - C-reactive protein and NT-proBNP as surrogate markers for pulmonary hypertension in Gaucher disease. AB - BACKGROUND: N-terminal pro-Brain Natriuretic Peptide (NT-proBNP) values correlate with mild-moderate pulmonary hypertension assessed by tricuspid insufficiency (TI) gradient>or=30 mm Hg in Gaucher disease. The purpose of this study is to ascertain improved risk stratification based on correlation with NT-proBNP and C reactive protein (CRP), a standard marker of inflammation. METHODS: Patients with type I Gaucher disease were selected to reflect differing degrees of echocardiographically determined TI gradient values. NT-proBNP was performed by immunoassay and CRP by standard methods. FINDINGS: There were 45 patients (18 males; 40%); mean age=42.5 (range: 4-80) years. Median NT-proBNP value=153 (range: 46-6703) pg/ml; median CRP value=0.145 (range: 0.02-2.69) mg/dl. There was a statistically significant correlation between these values (r=0.445, P<0.01). Elevations of CRP and NT-proBNP were risk factors for pulmonary hypertension with odds ratios of 8.47 and 4.9, respectively. The area under the Receiving Operator Characteristic (ROC) curve for diagnosis of pulmonary hypertension was 0.93+/-0.04 for CRP, and 0.7+/-0.1 for NT-proBNP. All patients with pulmonary hypertension had elevation of either CRP or NT-proBNP (100% sensitivity). CONCLUSIONS: Elevated CRP was a better predictor of pulmonary hypertension in Gaucher disease than elevated NT-proBNP values. Elevated CRP (>0.5 mg/dl) or elevated NT-proBNP (>150 pg/ml) reduces the need to perform echocardiography by more than half, even in this group with over-representation of pulmonary hypertension. The role of inflammatory features in pulmonary hypertension in Gaucher disease is discussed. Further studies are required to ascertain if this approach is useful for prognosis of pulmonary hypertension. PMID- 15885603 TI - Exosomes and the MICA-NKG2D system in cancer. AB - Exosomes are nanometer sized vesicles, secreted by a diverse range of cell types, whose biological functions remain ambiguous. Several groups have demonstrated the potential of manipulating exosomes for activating cellular immune responses. The possibility that exosomes may inhibit immunological responses, however, has not been widely addressed. We have investigated if exosomes produced by tumor cells can inhibit immunological functions, through modulating expression of the NKG2D receptor by effector cells. Incubating tumor exosomes with fresh peripheral blood leukocytes resulted in a marked reduction in the proportion of NKG2D-positive CD3+CD8+ Cells, and CD3- cells by 48 h. This effect was dose dependent and was shown with exosomes from different tumor cells including breast cancer and mesothelioma. Analysis of tumor exosome-phenotype revealed positive expression of several NKG2D ligands, and antibody blocking experiments revealed the importance of such ligands in driving the reduction in the proportion of NKG2D-positive effector cells. The functional importance of the decrease in NKG2D-positive cells was addressed in vitro cytotoxicity assays. For example a CD8+ T cell line pre incubated with tumor exosomes had significant decreased capacity to kill peptide pulsed T2 target cells. These data highlight a role for tumor exosomes bearing NKG2D ligands as a mechanism contributing to cancer immune evasion. PMID- 15885604 TI - Revisiting the road to the discovery of exosomes. AB - A look-back in time is presented to review the 'Alice in Blunderland' approach which led to the discovery of exosomes in and their extrusion from maturing reticulocytes. The key experiments which established that these structures are naturally occurring in both nucleated and non-nucleated reticulocytes are reviewed. The protein content of reticulocyte exosomes is largely that of plasma membrane proteins which are known to diminish in the course of reticulocyte maturation. Thus their protein content may vary depending on the species origin of the reticulocyte. To date, all exosomes also contain HSP 70, a major cellular chaperone which is not transmembrane bound. Significantly, the proteins externalized are intact and retain catalytic activity as well as native ligand binding activity. Their fate remains unknown. PMID- 15885605 TI - Suppression of eGFP expression in erythroid-specific transgenic mice by siRNA. AB - We have studied the role of small interfering RNA (siRNA) in the suppression of the enhanced green fluorescent protein (eGFP) in vitro and in vivo. siRNA plasmids for suppressing the eGFP expression were constructed, in which H1 promoter amplified from human 293 T cells was used to drive the small interfering RNA (19-nt) synthesis. In vitro studies demonstrated that the constructed siRNA plasmids had an effective suppressive effect on eGFP expression in 293 T and Mel cells. When the siRNA plasmid was injected into erythroid-specific eGFP transgenic mice, the eGFP expression were significantly suppressed (over 20% reduction) in the recipient mice compared to the control mice and the suppressing effect lasted for 2 weeks after one single injection. Moreover, the suppressive effect could be re-generated or boosted with secondary injections. Our results demonstrate a tissue-specific gene suppression by siRNA treatment. PMID- 15885606 TI - Iron overload in an African American woman with SS hemoglobinopathy and a promoter mutation in the X-linked erythroid-specific 5-aminolevulinate synthase (ALAS2) gene. AB - We report the case of an African American woman with sickle cell anemia and iron overload incompletely explained by erythrocyte transfusion who is heterozygous for a promoter mutation in the X-linked erythroid-specific 5-aminolevulinate synthase gene (ALAS2): a C to G transversion at nucleotide -206 from the transcription start site, as defined by primer extension (-258 from the start ATG). This mutation has previously been associated with sideroblastic anemia and iron overload in members of a Welsh kinship. No coding region mutation of HFE, FPN1, TFR2, HAMP, or HJV genes was detected. The mother of the proband has mild, chronic anemia and is also heterozygous for the same proximal promoter region mutation of ALAS2. However, she has no evidence of iron overload. We conclude that an ALAS2 promoter region mutation could partly account for iron overload in the present proband, and that this or other ALAS2 mutations could explain the occurrence of iron overload in other whites or blacks with or without anemia. The occurrence of anemia and iron overload may be discordant in women heterozygous for ALAS2 mutations. PMID- 15885607 TI - The homozygous state for Hb Crete [beta129 (H7) Ala-->Pro] is associated with a complex phenotype including erythrocytosis and functional anemia. AB - Hb Crete, an electrophoretically neutral, unstable, high oxygen affinity variant, was characterized by protein and DNA analyses in the homozygous state in a 32 year-old woman from Crete, with erythrocytosis and microcytosis. The proband and members of her family over 3 generations, including 5 carriers of Hb Crete, were subject to clinical, hematological and biochemical investigations, and DNA, RNA and protein studies were carried out. The proband demonstrated features associated with disturbed hemoglobin (Hb) structure and function, including erythrocytosis and additionally a state of functional anemia, the latter reflected by increased erythropoetin levels and cardiac output. In addition, all the carriers surprisingly had hematological and biosynthetic findings more usually associated with thalassemia trait. The structural change in Hb Crete only partly explains all the pathological manifestations of this variant, and other mechanisms are discussed. PMID- 15885608 TI - Selection with a regulated cell growth switch increases the likelihood of expression for a linked gamma-globin gene. AB - Several lines of evidence indicate that in vivo drug selection can be used to overcome the low rates of gene transfer and engraftment encountered in many hematopoietic stem cell gene therapy settings. However, whether selection imposed on one transcription cassette effects the likelihood of expression from a second, independent transcription cassette within the same vector has been less well studied. In order to address this issue, we engineered an oncoretrovirus vector to express two separate transcription units: (i) a bicistronic cassette encoding both GFP and a pharmacologically regulated cell growth switch based on the thrombopoietin receptor Mpl; and (ii) a highly position-dependent second cassette encoding human gamma-globin. Studies in cell cultures and in mice transplanted with transduced marrow indicated that selective expansion increased by more than 9-fold the fraction of erythroid cells expressing the linked but separate expression cassette for gamma-globin. This increase was far greater then that observed for the bicistronic GFP gene, and cannot be explained by a simple increase in the fraction of cells containing provirus. These results suggest that selective expansion favors erythroid stem/progenitor cells with provirus integrated at chromosomal sites which are relatively resistant to silencing position effects. PMID- 15885609 TI - Structural insights in platelet receptor synergism-antiplatelet therapy in post ischemic cerebrovascular events. AB - Synergy between agonists of platelet aggregation, namely, ADP and epinephrine, has been studied in patients having a history of cerebrovascular ischemic event. There is a significant variability of responsiveness among individuals towards clopidogrel, which is a specific inhibitor of the low-affinity human purinergic receptor (P2Y12). For responders of clopidogrel, simultaneous application of ADP and epinephrine at sub-threshold concentrations (i.e., concentration below the threshold concentration at which aggregation occurs) leads to platelet aggregation, which is followed by deaggregation. For non-responders of the drug, the synergism seems to be stronger, showing no deaggregatory pattern. The inhibition of synergism by yohimbine hydrochloride (YH), a blocker of alpha2A adrenoreceptors is more pronounced in non-responders. A simple structural model based on receptor-receptor interaction is proposed to explain the synergism. The model explains synergy in terms of cooperative interaction between the low affinity ADP receptor P2Y12 (Swiss Prot:Q9H244) and the alpha2A-adrenoreceptor (Swiss Prot:P08913). It follows that the synergistic effect can be achieved in only one of the two 3D structures for the alpha2A-adrenoreceptor P08913 permitted by homology modeling, as there is a better docking interface with the Q9H244. The synergism itself and the observed dichotomous phenomenon in relation to inhibition of synergism among responders and non-responders can be accounted for, if the interacting receptors on the dynamic membrane interface compete with the clopidogrel binding. PMID- 15885610 TI - Mutations in the reverse transcriptase component of telomerase (TERT) in patients with bone marrow failure. AB - Human telomerase has two core components, the RNA molecule (TERC) that provides the template for telomere repeat elongation and a reverse transcriptase (TERT) that is responsible for the addition of telomere repeats at the ends of each chromosome. Mutations in TERC have been found in the autosomal-dominant form of the inherited bone marrow failure syndrome dyskeratosis congenita and in a subset of patients with aplastic anemia and myelodysplasia. These patients have short telomeres compared to age-matched controls. These observations suggest that uncharacterised cases of dyskeratosis congenita/aplastic anemia may have mutations in TERT or other molecules that associate with TERC in the telomerase complex. We have therefore screened the TERT gene for mutation by denaturing HPLC in 80 patients with inherited and acquired bone marrow failure (24 with dyskeratosis congenita, 36 with constitutional aplastic anemia, 13 with idiopathic aplastic anemia and 7 with other forms of bone marrow failure). 15 different TERT mutations have been identified. Of these, 5 are in flanking intron sequences, 6 are synonymous and 4 are non-synonymous (missense) substitutions in the coding sequence. These are the first natural mutations of TERT to be described and we highlight their possible pathogenic role in the development of bone marrow failure. PMID- 15885611 TI - Temporal distribution of motorcyclist injuries and risk of fatalities in relation to age, helmet use, and riding while intoxicated in Khon Kaen, Thailand. AB - This study investigated the temporal distribution of risky behaviors among injured motorcyclists, that is, riding unhelmeted or while intoxicated, and showed how they are associated with risk of fatal injuries. Data of motorcyclists injured in Khon Kaen municipality in northeastern Thailand and transferred to Khon Kaen Regional Hospital were obtained from the trauma registry system of the hospital. Case fatalities were compared by time of day, age group, helmet use, and alcohol intoxication. Unhelmeted riding peaked late in the evening and riding while intoxicated peaked around midnight. Both were associated with increased fatality risk after stratification by time of day; the odds ratios were 3.49 (95% confidence interval (CI) = 1.48-9.36) and 3.01 (CI = 1.71-5.19), respectively. Nighttime injuries were not significantly associated with increased fatality risk after stratification by helmet use or alcohol intoxication. Unhelmeted driving was prevalent and associated with higher fatality risk among younger drivers, whereas intoxicated driving was less prevalent among teens but associated with increased risk among those aged 20-39 years. This study shows that riding unhelmeted or while intoxicated can explain the increased fatality risk at night, suggesting that safety education or enforcements should be targeted at specific age groups and appropriate times. PMID- 15885612 TI - Are speed enforcement cameras more effective than other speed management measures? The impact of speed management schemes on 30 mph roads. AB - This paper presents the results of an evaluation of the impact of various types of speed management schemes on both traffic speeds and accidents. The study controls for general trends in accidents, regression-to-mean effects and migration, separately estimating the accident changes attributable to the impact of the schemes on traffic speed and on traffic volume. It was found that, when judged in absolute terms, all types of speed management scheme have remarkably similar effects on accidents, with an average fall in personal injury accidents of about 1 accident/km/year. In terms of the percentage accident reduction, however, engineering schemes incorporating vertical deflections (such as speed humps or cushions) offer the largest benefits: at 44%, the average reduction in personal injury accidents attributable to such schemes, is twice that at sites where safety cameras were used to control speeds (22%) and they were the only type of scheme to have a significant impact on fatal and serious accidents. Other types of engineering scheme (with a fall of 29% in personal injury accidents) were on average less effective in reducing accidents than schemes with vertical features but more effective than cameras. All types of scheme were generally effective in reducing speeds, with the largest reductions tending to be obtained with vertical deflections and the smallest with other types of engineering schemes. PMID- 15885613 TI - Drug identification performance on the basis of observable signs and symptoms. AB - A double blind study was performed to evaluate the ability of trained police officers to detect drug impairments and to identify the type of drug responsible for the impairment, on the basis of observed symptoms and psychophysical measurements of performance alone. The officers were not allowed to interview the subjects, and their conclusions were based solely on observable signs and symptoms on systematically measured vital signs, and on standardized sobriety tests of motor coordination. Results showed that with this partial information, the officers are able to detect drug impairment at better-than-chance levels with a sensitivity (correct detection of impairments) of 72%, but with a specificity of 43% (false alarm rate of 57%). Furthermore, the association between drug ingestion and identification of the specific impairing drug category was not very high, with sensitivities ranging from a low of 10% for amphetamine to a high of 49% for cannabis. Based on both sensitivity and specificity, drug identification was best for alprazolam impairment, noticeably poorer for cannabis and codeine impairment, and no better than chance for amphetamine impairment. Performance could have been improved if the officers were to list the two most probable impairing drugs (rather than limit their decision to only one), and if they were more consistent in their interpretation of observable signs and symptoms. PMID- 15885614 TI - Identifying gene targets for the metabolic engineering of lycopene biosynthesis in Escherichia coli. AB - The identification of genetic targets that are effective in bringing about a desired phenotype change is still an open problem. While random gene knockouts have yielded improved strains in certain cases, it is also important to seek the guidance of cell-wide stoichiometric constraints in identifying promising gene knockout targets. To investigate these issues, we undertook a genome-wide stoichiometric flux balance analysis as an aid in discovering putative genes impacting network properties and cellular phenotype. Specifically, we calculated metabolic fluxes such as to optimize growth and then scanned the genome for single and multiple gene knockouts that yield improved product yield while maintaining acceptable overall growth rate. For the particular case of lycopene biosynthesis in Escherichia coli, we identified such targets that we subsequently tested experimentally by constructing the corresponding single, double and triple gene knockouts. While such strains are suggested (by the stoichiometric calculations) to increase precursor availability, this beneficial effect may be further impacted by kinetic and regulatory effects not captured by the stoichiometric model. For the case of lycopene biosynthesis, the so identified knockout targets yielded a triple knockout construct that exhibited a nearly 40% increase over an engineered, high producing parental strain. PMID- 15885615 TI - Improvement of nikkomycin production by enhanced copy of sanU and sanV in Streptomyces ansochromogenes and characterization of a novel glutamate mutase encoded by sanU and sanV. AB - Previous studies revealed that two genes-sanU and sanV were associated with nikkomycin biosynthesis in Streptomyces ansochromogenes. A plasmid used to increase an extra copy of sanU and sanV was constructed and introduced into wild type strain. HPLC results showed that nikkomycin production of recombinant strain was about 1.8 fold than that of wild-type strain. RT-PCR analysis indicated that the transcriptional level of sanU and sanV in this recombinant strain was about two folds than that of wild-type strain. The sanU and sanV were expressed in E. coli BL21 (DE3). SanU and SanV were purified individually. SanU and SanV assembled with coenzyme B12 to form a complete enzyme in vitro, which showed glutamate mutase activity. The glutamate mutase converted L-glutamate toL-threo beta-Methylaspartic acid, and then l-threo-beta-Methylaspartic acid was probably deaminated to form 2-oxo-3-methylsuccinic acid to join biosynthetic pathway of the peptidyl moiety HPHT in S. ansochromogenes. SanU is the coenzyme B12-binding component and more than two folds of SanU are required for maximal enzyme activity. The optimal pH and temperature for the formed enzyme are 7.5-8.5 and 35 42 degrees C, respectively. Sulfhydryl compounds are important for activity of the reassembled enzyme. PMID- 15885616 TI - Gas-inducible product gene expression in bioreactors. AB - Inducible transgene expression technologies are of unmatched potential for biopharmaceutical manufacturing of unstable, growth-impairing and cytotoxic proteins as well as conditional metabolic engineering to improve desired cell phenotypes. Currently available transgene dosing modalities which rely on physical parameters or small-molecule drugs for transgene fine-tuning compromise downstream processing and/or are difficult to implement technologically. The recently designed gas-inducible acetaldehyde-inducible regulation (AIR) technology takes advantage of gaseous acetaldehyde to modulate product gene expression levels. At regulation effective concentrations gaseous acetaldehyde is physiologically inert and approved as food additive by the Federal Drug Administration (FDA). During standard bioreactor operation, gaseous acetaldehyde could simply be administered using standard/existing gas supply tubing and eventually eliminated by stripping with inducer-free air. We have determined key parameters controlling acetaldehyde transfer in three types of bioreactors and designed a mass balance-based model for optimal product gene expression fine tuning using gaseous acetaldehyde. Operating a standard stirred-tank bioreactor set-up at 10 L scale we have validated AIR technology using CHO-K1-derived serum free suspension cultures transgenic for gas-inducible production of human interferon-beta (IFN-beta). Gaseous acetaldehyde-inducible IFN-beta production management was fully reversible while maintaining cell viability at over 95% during the entire process. Compatible with standard bioreactor design and downstream processing procedures AIR-based technology will foster novel opportunities for pilot and large-scale manufacturing of difficult-to-produce protein pharmaceuticals. PMID- 15885617 TI - Complementary identification of multiple flux distributions and multiple metabolic pathways. AB - Cell robustness and complexity have been recognized as unique features of biological systems. Such robustness and complexity of metabolic-reaction systems can be explored by discovering, or identifying, the multiple flux distributions (MFD) and redundant pathways that lead to a given external state; however, this is exceedingly cumbersome to accomplish. It is, therefore, highly desirable to establish an effective computational method for their identification, which, in turn, gives rise to a novel insight into the cellular function. An effective approach is proposed for complementarily identifying MFD in metabolic flux analysis and multiple metabolic pathways (MMP) in structural pathway analysis. This approach judiciously integrates flux balance analysis (FBA) based on linear programming and the graph-theoretic method for determining reaction pathways. A single metabolic pathway, with the concomitant flux distribution and the overall reaction manifesting itself as the desired phenotype under some environmental conditions, is determined by FBA from the initial candidate sequence of metabolic reactions. Subsequently, the graph-theoretic method recovers all feasible MMP and the corresponding MFD. The approach's efficacy is demonstrated by applying it to the in silico Escherichia coli model under various culture conditions. The resultant MMP and MFD attaining a unique external state reveal the surprising adaptability and robustness of the intricate cellular network as a key to cell survival against environmental or genetic changes. These results indicate that the proposed approach would be useful in facilitating drug discovery. PMID- 15885618 TI - Metabolic engineering of Escherichia coli for industrial production of glucosamine and N-acetylglucosamine. AB - Glucosamine and N-acetylglucosamine are currently produced by extraction and acid hydrolysis of chitin from shellfish waste. Production could be limited by the amount of raw material available and the product potentially carries the risk of shellfish protein contamination. Escherichia coli was modified by metabolic engineering to develop a fermentation process. Over-expression of glucosamine synthase (GlmS) and inactivation of catabolic genes increased glucosamine production by 15 fold, reaching 60 mg l(-1). Since GlmS is strongly inhibited by glucosamine-6-P, GlmS variants were generated via error-prone PCR and screened. Over-expression of an improved enzyme led to a glucosamine titer of 17 g l(-1). Rapid degradation of glucosamine and inhibitory effects of glucosamine and its degradation products on host cells limited further improvement. An alternative fermentation product, N-acetylglucosamine, is stable, non-inhibitory to the host and readily hydrolyzed to glucosamine under acidic conditions. Therefore, the glucosamine pathway was extended to N-acetylglucosamine by over-expressing a heterologous glucosamine-6-P N-acetyltransferase. Using a simple and low-cost fermentation process developed for this strain, over 110 g l(-1) of N acetylglucosamine was produced. PMID- 15885619 TI - Increased levels of erucic acid in Brassica carinata by co-suppression and antisense repression of the endogenous FAD2 gene. AB - Erucic acid and its derivatives represent important industrial feedstock compounds, and there is an increasing demand for the production of high erucate oils in this regard. Our goal therefore, is to develop high erucic acid (HEA) Brassicaceae lines with increased proportions of erucic acid and very long-chain fatty acids (VLCFAs). We proposed that oleate availability may be a rate-limiting factor in the biosynthesis of erucic acid. We have tried to address this question by manipulating the expression of the endogenous FAD2 gene in B. carinata using co-supression and antisense approaches. Both methods resulted in transgenic lines exhibiting decreased proportions of polyunsaturated C18 fatty acids (18:2+18:3) and concomitant and significantly increased proportions of 18:1, 22:1 and total VLCFAs. Co-suppressed FAD2 B. carinata lines exhibited 3-18% decreases in 18:2, 22-49% decreases in 18:3 and significantly increased proportions of 18:1 (36 99%), 22:1 (12-27%) and VLCFAs (6-15%). Transgenic B. carinata lines developed using an antisense FAD2 approach exhibited decreased proportions of 18:2 and 18:3 (9-39% and 33-48%, respectively) and significantly increased proportions of 18:1 (54-130%), 22:1 (5-19%) and VLCFAs (6-21%). The possibility of using these approaches to produce prototype transgenic germplasm of the Brassicaceae accumulating seed oils with improved proportions of erucic and other VLCFAs is discussed. PMID- 15885620 TI - Generation of serum-stabilized retroviruses: reduction of alpha1,3gal-epitope synthesis in a murine NIH3T3-derived packaging cell line by expression of chimeric glycosyltransferases. AB - Retroviral vectors released from mouse-derived packaging cell lines are inactivated in human sera by naturally occurring antibodies due to the recognition of Galalpha1,3Galbeta1,4GlcNAc (alphagal-epitope) decorated surface proteins. In this study, an extensive analysis of the glycosylation potential of NIH3T3-derived PA317 packaging cells using combined MALDI/TOF-MS and HPAE-PAD reveals that 34% of the N-glycan moiety represents alphagal-epitope containing structures. Stable expression of glycosyltransferases and transport signal chimeras has been demonstrated to represent an efficient tool to alter cell- and species-specific glycosylation (Grabenhorst and Conradt, 1999. J. Biol. Chem. 274, 36107-36116). In order to reduce alphagal-epitope synthesis selected chimeric glycosyltransferases were constructed by fusing Golgi-signal sequences for compartment-specific localization with the catalytic domain of alpha2,3 sialyltransferase (ST3). Stable expression of these constructs in these cells resulted in a significant reduced alphagal-epitope synthesis, and moreover, a release of retroviral vectors showing an up to 3.5-fold increase in serum stability. Thus, our results suggest that the stably transfected cells stably transfected with chimeric glycosyltransferases compete efficiently with endogenous alpha1,3-galactosyltransferase. This approach allows favored glycodesign and we anticipate the applicability of such improved retroviral vectors produced by glycosylation engineered host cells for in vivo gene therapy and, furthermore, suggest the therapeutic benefit of this technology for xenotransplantation. PMID- 15885621 TI - Novel pathway engineering design of the anaerobic central metabolic pathway in Escherichia coli to increase succinate yield and productivity. AB - A novel in vivo method of producing succinate has been developed. A genetically engineered Escherichia coli strain has been constructed to meet the NADH requirement and carbon demand to produce high quantities and yield of succinate by strategically implementing metabolic pathway alterations. Currently, the maximum theoretical succinate yield under strictly anaerobic conditions through the fermentative succinate biosynthesis pathway is limited to one mole per mole of glucose due to NADH limitation. The implemented strategic design involves the construction of a dual succinate synthesis route, which diverts required quantities of NADH through the traditional fermentative pathway and maximizes the carbon converted to succinate by balancing the carbon flux through the fermentative pathway and the glyoxylate pathway (which has less NADH requirement). The synthesis of succinate uses a combination of the two pathways to balance the NADH. Consequently, experimental results indicated that these combined pathways gave the most efficient conversion of glucose to succinate with the highest yield using only 1.25 moles of NADH per mole of succinate in contrast to the sole fermentative pathway, which uses 2 moles of NADH per mole of succinate. A recombinant E. coli strain, SBS550MG, was created by deactivating adhE, ldhA and ack-pta from the central metabolic pathway and by activating the glyoxylate pathway through the inactivation of iclR, which encodes a transcriptional repressor protein of the glyoxylate bypass. The inactivation of these genes in SBS550MG increased the succinate yield from glucose to about 1.6 mol/mol with an average anaerobic productivity rate of 10 mM/h (approximately 0.64 mM/h-OD600). This strain is capable of fermenting high concentrations of glucose in less than 24 h. Additional derepression of the glyxoylate pathway by inactivation of arcA, leading to a strain designated as SBS660MG, did not significantly increase the succinate yield and it decreased glucose consumption by 80%. It was also observed that an adhE, ldhA and ack-pta mutant designated as SBS990MG, was able to achieve a high succinate yield similar to SBS550MG when expressing a Bacillus subtilis NADH-insensitive citrate synthase from a plasmid. PMID- 15885623 TI - The nature of the parkinsonism-dementia complex and amyotrophic lateral sclerosis of Guam and magnesium deficiency. AB - The parkinsonism-dementia complex (PDC) and amyotrophic lateral sclerosis (ALS) were the fatal neurological diseases, showing very high incidence during 1950 1970 and dramatic decrease after 1970 on Guam. Through the research, the present author insisted that; (1) NFTs in Guam ALS patients are merely a background feature widely dispersed in the population, (2) Guam ALS and PDC are basically different diseases, and (3) Guam ALS occurs initially as classic ALS. As pathogeneses of the diseases, intake of low calcium (Ca) and magnesium (Mg) and high aluminum water and of some plant excitatory neurotoxin has been speculated. To elucidate the pathogenesis, the author performed an experiment exposing rats to low Ca and/or Mg intake for two generations, so as to follow the actual way of human living on the island, since several generations live continuously in the same environment. The study indicates that continuous low Mg intake for two generations induces exclusive loss of dopaminergic neurons in in rats, and may support the Mg hypothesis in the pathogenesis of PDC of Guam. PMID- 15885624 TI - Pathogenesis of levodopa-induced dyskinesia: focus on D1 and D3 dopamine receptors. AB - Involuntary movements, or dyskinesia, represent a debilitating complication of levodopa therapy for Parkinson's disease. Taking advantage of a monkey brain bank constituted to study the pathophysiology of levodopa-induced dyskinesia, we here report the changes affecting D1, D2 and D3 dopamine receptors within the striatum of four experimental groups of non-human primates: normal, parkinsonian, parkinsonian treated with levodopa without or with dyskinesia. We also report the possible role of arrestin and G protein-coupled receptor kinases. PMID- 15885625 TI - Pathogenesis of nigral cell death in Parkinson's disease. AB - Parkinson's disease (PD) is primarily a sporadic condition which results mainly from the death of dopaminergic neurons in the substantia nigra. Its etiology remains enigmatic while its pathogenesis begins to be understood as a multifactorial cascade of deleterious factors. As of yet, most insights into PD pathogenesis are derived from toxic models of PD and show that the earlier cellular perturbations arising in dopaminergic neurons include oxidative stress and energy crisis. These alterations, rather than killing neurons, trigger subsequent death-related molecular pathways including elements of apoptosis. The fate of dopaminergic neurons in PD may also be influenced by additional factors such as excitotoxicity, emanating from the increased glutamatergic input from the subthalamic nucleus to the substantia nigra, and the glial response that arises in the striatum and the substantia nigra. In rare instances, PD can be familial, and those genetic forms have also provided clues to the pathogenesis of nigrostriatal dopaminergic neuron death including abnormalities in the mechanisms of protein folding and degradation as well as mitochondrial function. Although more remains to be elucidated about the pathogenic cascade in PD, the compilation of all of the aforementioned alterations starts to shed light on why and how nigral dopaminergic neurons may degenerate in this prominent disease, that is PD. PMID- 15885626 TI - Controversy: is Parkinson's disease a single disease entity? Yes. AB - Parkinson's disease (PD) is a common, and in principle, sporadic, neurodegenerative disorder that occurs in adults. Pathological studies have revealed that in PD, nerve cell loss and Lewy bodies (LB) are distributed widely in the nervous system. Moreover, molecular pathology has made remarkable advances over the last several years, after the identification of alpha-synuclein gene abnormality in familial PD. Extensive pathological findings support the idea that PD is a single disease entity and that there are no cases of PD in which neurodegeneration occurs only in the substantia nigra and in which there are no LBs. PMID- 15885627 TI - A definition of Parkinson's disease. AB - We are now entering a period when the process of unraveling the puzzles of 'Parkinson's Disease' has generated a mass of information that does not fit into our traditional concepts. Having split off 'Progressive Supranuclear Palsy' and 'Multiple System Atrophy', we are faced with the dilemma of what to do with the various forms of Parkinsonism caused by a number of different mutations, and how to interpret pathological heterogeneity. Faced with these new questions, it is time to reconsider how 'Parkinson's Disease' is to be defined. PMID- 15885628 TI - Pathological substrate of dementia in Parkinson's disease--its relation to DLB and DLBD. AB - The pathological background of dementia in Parkinson's disease (PD) has been a matter of controversy. Recent studies have revealed the appearance of many Lewy bodies in the cerebral cortex of PD with dementia (PDD). This pathological change corresponds to the neocortical type, which was originally reported as diffuse Lewy body disease (DLBD), or a transitional type of dementia with Lewy bodies (DLB). Clinical differentiation between DLBD and the transitional type of Lewy body disease (LBD) is sometimes difficult because the symptoms overlap. Therefore, it is appropriate, at least at present, that DLB and PDD are used as clinical diagnoses and that DLBD and the transitional- and brainstem-types of LBD are used as neuropathological diagnoses. PMID- 15885629 TI - Dementia with Lewy bodies and Parkinson's disease with dementia: are they different? AB - The relationship of dementia with Lewy bodies (DLB) and Parkinson's disease with dementia (PDD) is debated. In DLB, dementia antedates Parkinsonism; in PDD, Parkinsonism antedates dementia. Other than presenting features, diagnostic measures fail to distinguish DLB from PDD. There are few or no pathologic differences between DLB and PDD. In most cases cortical Lewy bodies (LBs) are widespread and there is coexistent Alzheimer type pathology, insufficient to diagnose Alzheimer's disease. Given the predominance of Parkinsonism in PDD, neuronal loss in the substantia nigra is more severe in PDD than DLB. Further clinicopathologic studies are needed to define other pathologic differences between DLB and PDD and to explore the role of neuritic, basal forebrain and striatal pathology in these clinical syndromes. PMID- 15885630 TI - Neuroinflammatory processes in Parkinson's disease. AB - In Parkinson's disease (PD), post-mortem examination reveals a loss of dopaminergic (DA) neurons in the substantia nigra (SN) associated with a massive astrogliosis and the presence of activated microglial cells. Similarly, microglial activation has also been reported to be associated with the loss of DA neurons in animal models of PD induced by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), rotenone, annonacine and lipopolysaccharide (LPS). Recent evidence suggests that the disease may progress even when the initial cause of neuronal degeneration has disappeared, raising the possibility that toxic substances released by glial cells could be involved in the propagation of neuronal degeneration. Inhibition of the glial reaction and the inflammatory processes may thus represent a therapeutic target to reduce neuronal degeneration in PD. PMID- 15885631 TI - Immune reconstitution in HIV infection: how to measure thymic function? PMID- 15885632 TI - The fibroblast-like synovial cell in rheumatoid arthritis: a key player in inflammation and joint destruction. AB - Although multiple cell types are present in the rheumatoid joint, the fibroblast like synovial cell (FLS) is among the most prominent. It is now appreciated that the FLS is not only space-filling, but is directly responsible for cartilage destruction, and also drives both inflammation and autoimmunity. In this article, we consider the normal role of the FLS in healthy joints, and review evidence that implicates the FLS as a central player in the propagation of rheumatoid arthritis. PMID- 15885633 TI - Ruminating on rheumatoid arthritis and transplantation. PMID- 15885634 TI - Eliminating eosinophilic esophagitis. PMID- 15885635 TI - HLA class II involvement in HIV-associated Toxoplasmic encephalitis development. AB - A total of 220 individuals were included in this study, 112 HIV-seronegative healthy individuals and 108 HIV-1-infected patients involving: 18 AIDS patients with Toxoplasmic encephalitis (AIDS-TE), 49 AIDS patients without TE, and 41 asymptomatic patients, were genotyping for DR and DQ loci by molecular biology techniques. Fisher's Exact test was used for statistical analysis. HLA-DQB*0402 and DRB1*08 alleles were associated with a high risk to develop opportunistic infections with neurological involvement, mainly Toxoplasma encephalitis in relationship with subjects healthy (OR = 20.43; Pc = 7.0 x 10(-6) and OR = 11; Pc = 2.6 x 10(-4), respectively); in relationship with AIDS no TE (OR = 6.98; Pc = 0.028 and OR = 4.85; P = 0.012, Pc = 0.14) and with patients in asymptomatic stage (OR = 61.50, Pc = 8.4 x 10(-6) and OR = 19.38; Pc = 3.9 x 10(-4)), respectively. CONCLUSIONS: It was concluded that the presence of HLA-DQB*0402 and DRB1*08 alleles in HIV-1-positive patients could be considered risk factors for developing neurological opportunistic infections, mainly Toxoplasmic encephalitis. PMID- 15885636 TI - Multiparameter evaluation of human thymic function: interpretations and caveats. AB - After the provision of highly active antiretroviral therapy (HAART), the level of circulating CD4+ T cells increases in many adults infected with the human immunodeficiency virus, type 1 (HIV). To study factors involved in immune reconstitution, we have measured thymic abundance by CT scans, circulating naive phenotype CD4+ T cells by flow cytometry, and T cell receptor (TCR) rearrangement excision circles (TRECs) by quantitative PCR in 40 virologically suppressed, HIV infected adults and 33 age-matched, HIV-uninfected controls. In HIV-uninfected subjects, naive T cell numbers, thymic abundance, and the frequency of circulating naive CD4+ T cells bearing TRECs decreased with age, as expected. When corrected for this relationship with age, naive T cell numbers correlated significantly with naive T cell TREC frequencies. Virologically suppressed HIV infected subjects had higher TREC frequencies, and subjects over the age of 39 were more likely to have abundant thymus compared to age-matched, HIV-uninfected adults. Nevertheless, all HIV-infected subjects had reduced absolute numbers of naive T cells, irrespective of thymic size, age, or TREC frequencies. These data illustrate the complex relationship between these measures of thymic size and function and underscore the need to develop more definitive measures of thymic function in the future. PMID- 15885637 TI - Deficient IL-12 and dendritic cell function in common variable immune deficiency. AB - Patients with common variable immune deficiency have reduced serum IgG, IgA, and/or IgM, defective antibody production, and many have cellular abnormalities, including proliferative defects, accelerated T cell apoptosis, and insufficient production of IL-2 and IL-10. Excess monocyte intracellular IL-12 leading to a polarized Th-1-type response which could prevent antibody production has been suggested. Here we found that dendritic cells (DCs) of CVID subjects have a significantly reduced capacity to secrete IL-12, as compared to DCs of normal subjects when cultured with physiologic simulators: LPS (P = 0.0005), TNF-alpha (P = 0.006), or CD40-L fusion protein (P = 0.0004). CVID TNF-alpha or CD40-Ligand matured DCs were also significantly impaired in antigen presentation in mixed lymphocyte culture. Deficient IL-12 production was closely correlated to lymphocyte functions in vitro and to the absolute numbers of CD4 T cells in peripheral blood. While CVID DCs appear morphologically similar to DCs of normal subjects, the lack of IL-12 production and defective antigen presentation demonstrate functional defects. Deficient DC function could lead to attenuated T cell activation and defective immunization, features characteristic of CVID. PMID- 15885638 TI - Correlates of lymphoproliferative responses to measles, mumps, and rubella (MMR) virus vaccines following MMR-II vaccination in healthy children. AB - Cell-mediated immunity (CMI) to measles, mumps, and rubella viral antigens plays a critical role in providing long-term protection against these infectious diseases. We examined CMI by measuring lymphoproliferative response induced in response to stimulation with the above three antigens following two doses of measles, mumps, and rubella-II (MMR-II) vaccine in a randomly selected, population-based cohort of healthy children. We determined if a correlative and predictive intraclass relationship exists between CMI to the three components of MMR-II. We detected positive lymphoproliferative responses to measles, mumps, and rubella vaccines. Mumps vaccine used as an antigen had the highest median stimulation index followed by measles and rubella vaccines. The overall intraclass correlation value for lymphoproliferative response to measles, mumps, and rubella using Pearson's correlation was 0.61 (95% confidence interval = 0.56, 0.66). We observed a significant pairwise association to individual vaccine components between subjects in the upper and lower 10th percentile of immune response. This study demonstrates recall CMI post-MMR-II vaccination with significant intraclass correlation among the CMI responses to the three vaccine components. PMID- 15885639 TI - Ethinyl estradiol treats collagen-induced arthritis in DBA/1LacJ mice by inhibiting the production of TNF-alpha and IL-1beta. AB - We previously demonstrated the therapeutic effects of ethinyl estradiol (EE), an orally active estrogen and a component of birth control pills, in encephalitogenic autoimmune encephalomyelitis (EAE). In this study, we report the effectiveness of EE in treating collagen-induced arthritis (CIA) induced with bovine type II collagen (bCII) in DBA/1LacJ mice, a CIA susceptible strain. Both low and high doses of EE notably suppressed clinical and histological signs of CIA in a dose-dependent manner compared to vehicle-treated controls. Oral treatment with EE decreased proliferation and secretion of pro-inflammatory factors, TNF-alpha IFN-gamma, MCP-1 and IL-6 by bCII peptide-specific T cells, production of bCII-specific IgG2a antibodies, and mRNA for cytokines, chemokines and chemokine receptors in joint tissue. This is the first report demonstrating effective treatment of joint inflammation and clinical signs of CIA with orally administered ethinyl estradiol, thus supporting its possible clinical use for treating rheumatoid arthritis in humans. PMID- 15885640 TI - C-reactive protein concentration is not related to islet autoimmunity status in offspring of parents with type 1 diabetes. AB - Autoimmunity may be associated with acute or chronic inflammation. In order to determine whether the inflammatory marker C-reactive protein (CRP) was an indicator of inflammatory events that precede, predict, or associate with islet autoimmunity or type 1 diabetes, CRP was measured in sequential antibody negative, seroconversion, and follow-up-positive samples from 65 prospectively studied islet autoantibody-positive children. Although changes in CRP concentrations were observed in some children, overall CRP concentrations were similar in antibody-negative samples (median, 0.21 mg/L), antibody-positive samples (median, 0.26 mg/L), and samples at seroconversion (median, 0.26 mg/L). CRP concentrations at diabetes onset (median, 0.59 mg/L) were not significantly increased over antibody-negative samples (P = 0.07). CRP concentrations did not predict diabetes development. CRP concentrations were related to age (r = 0.26; P < 0.001) and were increased in samples obtained from October to January (P < 0.001). These findings suggest that CRP concentrations are not a valuable marker of progression to type 1 diabetes and highlight the importance of correcting analyses for seasonal variations. PMID- 15885641 TI - Effects of allergen inhalation and oral glucocorticoid on concentrations of serum soluble CD86 in allergic asthmatics. AB - The aim of the present study was to investigate effects of allergen inhalation and oral glucocorticoid on concentration of serum soluble CD86 in patients with allergic asthma. Our results showed that the serum soluble CD86 concentrations in the dual responder group increased from 491.8 +/- 15.4 IU/ml before allergen inhalation to 603.8 +/- 19.3 IU/ml 24 h after allergen inhalation. In the isolated early responders, there was no significant increase in serum soluble CD86 concentrations after allergen inhalation compared with baseline levels. There was a significant decrease in serum soluble CD86 concentrations after 2 weeks of glucocorticoid therapy (448.3 +/- 15.1 IU/ml) compared with baseline values (532.7 +/- 12.3 IU/ml), whereas there was no significant difference in the placebo group. This study has demonstrated that serum soluble CD86 concentrations increased after allergen inhalation in sensitized asthmatic subjects, and that serum sCD86 concentrations were downregulated by prednisolone therapy. PMID- 15885642 TI - PD-L1 is expressed by human renal tubular epithelial cells and suppresses T cell cytokine synthesis. AB - T cell activation is affected by both stimulatory and inhibitory co-signaling. MHC class II-expressing renal tubular epithelial cells (TEC) can function as APC for T cells. To study the influence of inhibitory ligands on TEC-mediated T cell activation, we examined the expression of programmed death ligand-1 (PD-L1) on human TEC line HK-2 cells, as well as in normal and diseased kidney samples. RT PCR, FACS, and immunocytochemistry showed that PD-L1 is constitutively expressed on HK-2 cells, and is dramatically upregulated by IFN-gamma. In situ hybridization and immunohistochemical staining revealed constitutive low expression of PD-L1 on proximal tubules at both mRNA and protein levels in normal kidneys, but much higher expression in kidneys with type IV lupus nephritis. In vitro, pretreatment of IFN-gamma-stimulated HK-2 cells with anti-PD-L1 significantly enhanced IL-2 secretion from cocultured, mitogen-activated Jurkat or human peripheral blood T cells. These results suggest that the PD-L1:PD-1 pathway negatively regulates T cell activation by TEC, and may play an inhibitory role in TEC-mediated immune activation and immunopathology in the kidney. PMID- 15885643 TI - Increase of activated T-cells and up-regulation of Smad7 without elevation of TGF beta expression in tonsils from patients with pustulosis palmaris et plantaris. AB - Pustulosis palmaris et plantaris (PPP) is known to be a skin disease related to tonsillitis, because the pustulosis often become exacerbated during acute tonsillitis and disappears after tonsillectomy. However, etiology of PPP remains unclear. In this study, we investigated the activation of tonsillar T-cell from PPP patients. Furthermore, we analyzed expressions of cytotoxic T-lymphocyte antigen-4 (CTLA4) that is a co-stimulatory molecule for inhibition of T-cell activation and of Smad7 that is a regulatory factor of TGF-beta intracellular signaling. For 47 Japanese patients with PPP who had tonsillectomy, the skin lesion was improved in 87% of PPP patient at 12 months after tonsillectomy. In quantitative immunohistologic analysis, T-cell nodules on tonsillar tissues from PPP patients were more expanded than those from the patients with obstructive sleep apnea syndrome (OSAS) (P = 0.015), and there was a positive correlation between the enlargement and clinical improvement (r = 0.422, P = 0.021). Flow cytometric analysis showed that the numbers of CD4+CD25+ and CD4+CD29+ cells in tonsils from PPP patients increased significantly compared to those from OSAS patients (P = 0.017, P = 0.016, respectively). Using reverse transcription polymerase chain reaction (RT-PCR) and Western blotting analyses with CD3+ tonsillar lymphocytes, we found that both expressions of Smad7 mRNA and protein were enhanced in PPP patients compared with OSAS patients (P = 0.03, P = 0.02, respectively), but expression of TGF-beta mRNA was not different between 2 groups. Although mRNA expression of CTLA4 was reduced in PPP patients compared with OSAS patients (P = 0.04), the CTLA4 surface protein expression was not different between 2 groups. These data suggest that helper T-cells are frequently activated in tonsils from PPP patients, and this activation may be related to unresponsiveness of TGF-beta1 by overexpression of Smad7. Such hyper-activation of T-cell may increase the risk of elicitation of self-reactive T-cell, being associated with pathogenesis of PPP. PMID- 15885644 TI - Armed CD4+ Th1 effector cells and activated macrophages participate in bile duct injury in murine biliary atresia. AB - Biliary atresia (BA) is an inflammatory cholangiopathy of infancy. A proposed mechanism regarding the pathogenesis of BA is that of a virus-induced, immune mediated injury to bile ducts. The rotavirus (RRV)-induced murine model of BA was utilized to determine the hepatic inflammatory response related to ductal obstruction and if the immune response recapitulated human BA. One week after infection, there was a significant increase in liver CD4(+) T cells producing IFN gamma and in macrophages producing TNF-alpha. The intrahepatic pattern of inflammation evolved rapidly from an initial predominant CD4(+) Th1 cellular response to a subsequent influx of activated macrophages producing TNF-alpha and iNOS. This immune response persisted despite viral clearance and was representative of the hepatic immune profile present in human BA. Utilization of the murine model of BA yielded mechanistic data that can provide much needed insight into the role played by different arms of the immune system related to the pathogenesis of human BA. PMID- 15885645 TI - Differential cytokine profile in children with cystic fibrosis. AB - The previously observed occurrence of antineutrophil cytoplasmic autoantibodies (ANCA) in patients who have cystic fibrosis (CF), together with the reported decrease in IgG2, a Th1-controlled isotype, suggests a potential for Th1/Th2 imbalance in CF patients with a possible Th2 predominance. 48 CF patients and 16 controls had levels of IFNgamma, IL-4, and IL-10 measured in supernatants of whole blood cell cultures stimulated by lipopolysaccharide (LPS) and phytohemaglutinine (PHA). The patients were divided into 2 groups: "low responders", having negligible secretion of cytokines (IFNgamma: 10.0-200.0 pg/ml, IL-4: 0.0-0.3 pg/ml) and "high responders", producing high levels of both IFNgamma (500.0-2000.0 pg/ml) and IL-4 (1.0-200.0 pg/ml). There was a statistically significant (P < 0.01) deterioration of lung function measured by an FEV(1) decline by 11.2% over 3 years in the "low responder" group. 10 of 16 "low responders" had chronic lung infections with P. aeruginosa while such infection was less prevalent in the "high responder" group where only 13 of 32 CF patients had positive cultures. A shift towards Th2 response was observed in the "high responder" group as children chronically infected with P. aeruginosa had greater IL-4 production than non-infected CF patients within the same cohort. ANCA autoantibodies were found only in the "high responder" group. Th2 immune response predominance in a subset of CF patients is associated with chronic P. aeruginosa infection. PMID- 15885646 TI - IL-6 levels predict disease variant and extent of organ involvement in patients with mastocytosis. AB - Mastocytosis is often associated with organ involvement and hematological disorders. Patients may also exhibit elevated levels of plasma IL-6. To gain insight into the relevance of this observation, we correlated plasma levels of IL 6 and soluble IL-6 receptor (sIL-6R) with multiple disease parameters in 29 patients with mastocytosis. Mean plasma IL-6 levels were elevated in patients compared to healthy controls (P < 0.0001). Disease category significantly correlated with plasma IL-6 levels, as did severity of bone marrow pathology, organomegaly, and extent of skin involvement. In plasma, there was a positive correlation of IL-6 to total tryptase, alkaline phosphatase, IgM, white blood cell count, prothrombin time, partial thromboplastin time, and neutrophil numbers. There was an inverse correlation to hemoglobin. sIL-6R levels were not elevated. These observations demonstrate that IL-6 is a useful surrogate marker of severity of hematologic disease and suggest that IL-6 contributes to pathology. PMID- 15885652 TI - A high conductance cationic channel from Phaseolus vulgaris roots incorporated into planar lipid bilayers. AB - A previously undescribed plasma membrane cation channel from Phaseolus vulgaris bean roots was studied after its incorporation into planar lipid bilayers. The channel allows the passage of monovalent cations excluding the flux of both anions (Cl-) and divalent cations (Ca2+). The channel presents a high ( approximately 213 pS) conductance in (300 mM Kcis+)/ (150 mMKtrans+) conditions. The probability of opening (Po) is low at all the tested voltages, but it increases significantly at trans-negative potentials. Permeability ratios (Pcation/PK+) under bi-ionic conditions follow the sequence: K+ (1.0)>NH4+ (0.86)>Na+ (0.78). Under the same conditions, the conductance ratios (gamma cation/gamma K+) follow the sequence: NH4+ (1.1) > or = K+ (1.0)>Na+ (0.80). The low probability of opening exhibited by the channel upon its incorporation into a lipid bilayer makes it a candidate to regulation by (and therefore participation in) cellular signalling networks. PMID- 15885653 TI - Cys351 and Cys361 of the Na+/glucose cotransporter are important for both function and cell-surface expression. AB - Here, we identify Cys351 and Cys361 as novel residues critical for the function and plasma membrane targeting of the Na+/glucose transporter-1 (SGLT1). HEK-293 cells expressing the C351A and C361A mutants showed no detectable Na(+)-coupled uptake for alpha-methyl glucoside (AMG). Cell-surface biotinylation and Western blot revealed that the two mutants were overexpressed in 293 cells; however, none of them exhibited normal cell-surface expression. When reconstituted in proteoliposomes, mutant SGLT1s demonstrated significantly lower affinity for AMG compared with the wild-type transporter. Incubation with the reducing agent dithiothreitol did not alter the catalytic activity of wild-type protein, but surprisingly, it nearly restored the ability of SGLT1-C351A and -C361A to bind and translocate AMG. Thus, the C351A and C361A mutations might cause a global reorganization of the disulfide bonds of SGLT1. Furthermore, we showed that a double mutation (C351A/C361A) restored the cell-surface expression of the single C-to-A mutants (C351A and C361A). PMID- 15885654 TI - Classical fear conditioning in the anxiety disorders: a meta-analysis. AB - Fear conditioning represents the process by which a neutral stimulus comes to evoke fear following its repeated pairing with an aversive stimulus. Although fear conditioning has long been considered a central pathogenic mechanism in anxiety disorders, studies employing lab-based conditioning paradigms provide inconsistent support for this idea. A quantitative review of 20 such studies, representing fear-learning scores for 453 anxiety patients and 455 healthy controls, was conducted to verify the aggregated result of this literature and to assess the moderating influences of study characteristics. Results point to modest increases in both acquisition of fear learning and conditioned responding during extinction among anxiety patients. Importantly, these patient-control differences are not apparent when looking at discrimination studies alone and primarily emerge from studies employing simple, single-cue paradigms where only danger cues are presented and no inhibition of fear to safety cues is required. PMID- 15885655 TI - I don't like it because it eats sprouts: conditioning preferences in children. AB - Although little is known about how preferences develop in childhood, work in adults suggests that evaluative responses to stimuli can be acquired through classical conditioning. In two experiments children were exposed to novel cartoon characters, that were either consistently paired with a picture of a disliked food (Brussels sprouts) or a liked food (ice cream). Relative preferences for these stimuli (and others) were measured before and after these paired presentations (Experiment 1): preferences for the cartoon character paired with Brussels sprouts decreased, whereas preferences for the character paired with ice cream increased. These preferences persisted after 10 un-reinforced trials. Experiment 2 replicated this finding using affective priming as an index of preference for the cartoon characters. These findings demonstrate that preferences to novel stimuli can be conditioned in children and result from associations formed between the stimulus and a stimulus possessing positive or negative valence. PMID- 15885656 TI - Differential CB1 and CB2 cannabinoid receptor-inotropic response of rat isolated atria: endogenous signal transduction pathways. AB - In this study, we have determined the contractile effects of CB1 and CB2 cannabinoid receptor activation on rat isolated atria and the different signaling pathways involved. Anandamide did not has significantly effect on atria contractility, however, the treatment with both CB1 (AM251) or CB2 (AM630) receptor antagonists, the endocannabinoids triggered stimulation or inhibition on contractility respectively. The ACEA stimulation of CB1 receptor exerted decrease on contractility, that significantly correlated with the decrement of cAMP and the stimulation of nitric oxide synthase (NOS) and the accumulation of cyclic GMP (cGMP). On the contrary, JWH 015 stimulation of CB2 receptor triggered positive contractile response that significantly correlated with the increase cAMP production. The inhibiton of adenylate cyclase activity impaired the JWH 015 activation of CB1 receptor induced positive contractile effect, while inhibitors of phospholipase C (PLC), NOS and soluble nitric oxide (NO)-sensitive guanylate cyclase blocked the dose-response curves of ACEA on contractility. Those inhibitors also attenuated the CB1 receptor-dependent increase in activation of NOS and cGMP accumulation. These results suggest that CB2 receptor agonist mediated positive contractile effect associated with increased production on cAMP while CB1 receptor agonist mediated decrease on contractility associated with decreased cAMP accumulation and increase production of NO and cGMP; that occur secondarily to stimulation of PLC, NOS and soluble guanylate cyclase. Data give pharmacological evidence for the existence of functional CB1 and CB2 cannabinoid receptors in rat isolated atria and may contribute to a better understanding the effects of cannabinoids in the cardiovascular system. PMID- 15885657 TI - Transfection of adult primary rat hepatocytes in culture. AB - The use of adult primary hepatocytes in culture is of importance for the understanding of hepatic processes at the cellular and molecular levels, and the possibility to employ transient transfection of reporter constructs is invaluable for mechanistic studies on hepatic gene regulation. Although frequently used, there is a lack of reports addressing optimization and characterization of transfection of primary rodent hepatocytes. Here, we have shown that the efficiency of biochemical transfection reagents varies significantly and that Lipofectamine2000 was a superior transfection reagent for adult primary rat hepatocytes when using luciferase reporter vectors. The efficiency increased when the cells were allowed ample time to adapt to the in vitro milieu. Cotransfection of a second reporter gene indicated a risk for promoter competition, and we found that relating reporter activity to total cellular protein content gave consistent and reliable results. Differentiation of the cells, achieved by including biomatrix from the Engelbreth-Holm-Swarm mouse sarcoma in the culture system, was to a larger extent required for hormonal/drug responses of transfected constructs than for responses of endogenous genes and assured responses of transfected constructs. Dexamethasone (Dex) is most often included in hepatocyte culture media, but we could not demonstrate a general beneficial effect of Dex on expression of luciferease reporter contructs. Using the established protocol, we have demonstrated responses of transfected constructs to growth hormone, glucocorticoid and LXR stimuli. PMID- 15885658 TI - Inhibition of multidrug resistance proteins MRP1 and MRP2 by a series of alpha,beta-unsaturated carbonyl compounds. AB - To study the possible interplay between glutathione metabolism of and MRP inhibition by thiol reactive compounds, the interactions of a series of alpha,beta-unsaturated carbonyl compounds with multidrug resistance proteins 1 and 2 (MRP1/ABCC1 and MRP2/ABCC2) were studied. Alpha,beta-unsaturated carbonyl compounds react with glutathione, and therefore either their parent compound or their intracellularly formed glutathione metabolite(s) can modulate MRP-activity. Inhibition was studied in Madin-Darby canine kidney cells stably expressing MRP1 or MRP2, and isolated Sf9-MRP1 or Sf9-MRP2 membrane vesicles. In the latter model system metabolism is not an issue. Of the series tested, three distinct groups could be discriminated based on differences in interplay of glutathione metabolism with MRP1 inhibition. Curcumin inhibited MRP1 transport only in the vesicle model pointing at inhibition by the parent compound. The glutathione conjugates of curcumin also inhibit MRP1 mediated transport, but to a much lesser extent than the parent compound curcumin. In the cellular model system, it was demonstrated that glutathione conjugation of curcumin leads to inactivation of its inhibitory potential. Demethoxycurcumin and bisdemethoxycurcumin inhibited MRP1 in both the vesicle and cellular model pointing at inhibitory potency of at least the parent compound and possibly their metabolites. A second group, including caffeic acid phenethyl ester inhibited MRP1-mediated calcein transport only in the MDCKII-MRP1 cells, and not in the vesicle model indicating that metabolism appeared a prerequisite to generate the active inhibitor. Finally cinnamaldehyde, crotonaldehyde, trans-2-hexanal, citral, and acrolein did not inhibit MRP1. For MRP2, inhibition was much less in both model systems, with the three curcuminoids being the most effective. The results of this study show the importance to study the complex interplay between MRP-inhibitors and their cellular metabolism, the latter affecting the ultimate potential of a compound for cellular MRP-inhibition. PMID- 15885659 TI - Metabolic activation of zebularine, a novel DNA methylation inhibitor, in human bladder carcinoma cells. AB - Zebularine (2(1H)-pyrimidinone riboside, Zeb), a synthetic analogue of cytidine that is a potent inhibitor of cytidine deaminase, has been recently identified as a general inhibitor of DNA methylation. This inhibition of DNA methyltransferase (DNMT) is hypothesized to be mechanism-based and result from formation of a covalent complex between the enzyme and zebularine-substituted DNA. Metabolic activation of Zeb thus requires that it be phosphorylated and incorporated into DNA. We have quantitatively assessed the phosphorylation and DNA incorporation of Zeb in T24 cells using 2-[(14)C]-Zeb in conjunction with gradient anion-exchange HPLC and selected enzymatic and spectroscopic analyses. The corresponding 5'-mono , di- and triphosphates of Zeb were readily formed in a dose- and time-dependent manner. Two additional Zeb-containing metabolites were tentatively identified as diphosphocholine (Zeb-DP-Chol) and diphosphoethanolamine adducts. Intracellular concentrations of Zeb-TP and Zeb-DP-Chol were similar and greatly exceeded those of other metabolites. DNA incorporation occurred but was surpassed by that of RNA by at least seven-fold. Equivalent levels and similar intracellular metabolic patterns were also observed in the Molt-4 (human T-lymphoblasts) and MC38 (murine colon carcinoma) cell lines. For male BALB/c nu/nu mice implanted s.c. with the EJ6 variant of T24 bladder carcinoma and treated i.p. with 500mg/kg 2-[(14)C] Zeb, the in vivo phosphorylation pattern of Zeb in tumor tissue examined 24h after drug administration was similar to that observed in vitro. The complex metabolism of Zeb and its limited DNA incorporation suggest that these are the reasons why it is less potent than either 5-azacytidine or 5-aza-2'-deoxycytidine and requires higher doses for equivalent inhibition of DNMT. PMID- 15885660 TI - Galphaq-coupled receptor signaling enhances adenylate cyclase type 6 activation. AB - Calcium signaling robustly inhibits AC6 activity in membrane preparations and in intact cells via capacitative calcium entry (CCE). However, the release of intracellular calcium has not been demonstrated to robustly alter AC6 signaling and activation of Galpha(q)-coupled receptors in tissues that express AC6 enhances cyclic AMP accumulation. To specifically examine the ability of Galpha(q)-coupled receptors to modulate AC6 signaling in intact cells, we used stably transfected HEK-AC6 cells. We demonstrate that AC6 activation is potentiated by activation of endogenous muscarinic receptors expressed in HEK293 cells. Muscarinic receptor activation failed to potentiate the activation of the closely related AC5 isoform. Expression of recombinant Galpha(q)-coupled muscarinic or serotonin receptors, or constitutively active Galpha(q), also potentiated drug-stimulated cyclic AMP accumulation in HEK-AC6 cells. Muscarinic receptor-mediated potentiation of AC6 activation was not due to activation of PKC or modulation of Galpha(i/o)-mediated inhibition of AC6. We demonstrate that calcium chelation or inhibition of calmodulin attenuates the effect of carbachol on AC6 activation. These data support the hypothesis that Galpha(q)-coupled receptor-mediated calcium signaling potentiates AC6 activation in intact cells. PMID- 15885661 TI - Intestinal inflammation induces adaptation of P-glycoprotein expression and activity. AB - Recent studies suggest that P-glycoprotein (Pgp) encoded by MDR1 gene, may be an important factor in the pathogenesis of inflammatory bowel disease (IBD). In this study, we investigated intestinal Pgp expression and activity: (1) in IL10 deficient (IL10(-/-)) mice which spontaneously develop intestinal inflammation affecting the small and large intestine and (2) in DSS (dextran sodium sulfate) induced rat colitis. In IL10(-/-) enterocolitis mice, rhodamine 123 efflux was reduced by two to three-fold along the small and large intestine. This decrease was associated with a reduction in membrane's Pgp protein levels. A similar three fold decrease in Pgps activity and expression was observed in the proximal colon in DSS-induced colitis in rats. However, in the non-inflamed ileum in DSS-induced rat colitis, epithelial cell's Pgp activity and protein levels were unexpectedly increased. This effect was specific to local inflammation since LPS induced systemic inflammation did affect neither the intestinal rho 123 efflux transport nor the abundance of the Pgp protein. These data demonstrate for the first time, an impaired function of epithelial Pgp in IL10 deficient enterocolitis mice. They also show an increase in Pgps activity in the non-inflamed ileum in the DSS induced rat colitis, which may represent an adaptive mechanism to compensate the impaired activity of Pgp in the colon. PMID- 15885662 TI - Combining exercise and cyclooxygenase-2 inhibition does not ameliorate learning deficits after brain insult, despite an increase in BDNF levels. AB - Neurodegeneration can produce behavioral impairments. Previously, we have found that inhibition of cyclooxygenase-2 activity or physical activity was neuroprotective during kainic-acid-induced neural loss. Here, we investigated the combined effect of exercise pre-insult and cyclooxygenase inhibitor treatment post-kainate-induced brain damage. However, in spite of an increase in BDNF levels, the combination did not improve behavioral performance in Morris watermaze and object exploration tasks. PMID- 15885663 TI - Substance P does not play a critical role in neurogenic inflammation in the rat masseter muscle. AB - In this study, we performed a series of experiments to investigate whether substance P (SP) contributes to neurogenic inflammation in the skeletal muscle tissue. Intramuscular injection of an inflammatory irritant, mustard oil (MO), induces significant edema formation in the rat masseter muscle. In order to study the contribution of endogenous SP in the MO-induced edema, groups of rats were pretreated with two different doses (100 nmol; 1 microl) of either peptidergic (Sendide) or non-peptidergic (L703, 606) neurokinin 1 (NK1) receptor antagonist in one masseter muscle 15 min prior to the MO injection in the same muscle. The extent of edema was assessed as the percent weight difference of the injected muscle compared to the non-injected muscle. Neither Sendide nor L703,606 pretreatment resulted in a significant inhibition of the MO-induced edema in the masseter muscle. Exogenous application of SP also produced a significant swelling of the muscle, which was blocked by L703,606 (1 microl) pretreatment, suggesting that evoked release of SP following MO injection is not sufficient to induce significant edema formation. Capsaicin (1% in 25 microl), which is known to cause neurogenic inflammation, failed to produce edema formation in the masseter muscle. The same concentration of capsaicin injected into the hindpaw produced significant swelling of the injected paw. Taken together, these results provide compelling evidence that, unlike cutaneous or joint tissue, SP does not play a critical role in inducing neurogenic inflammation in the skeletal muscle tissue. PMID- 15885664 TI - Calbindin D-28k is expressed in the microvascular basal lamina in the ventral horn at early time after transient spinal cord ischemia in the rabbit. AB - Much evidence has been accumulated that the increased expression of calbindin D 28k (CB) is involved in the blockade of calcium-evoked excitotoxicity in cerebral ischemia. We investigated the expression of CB in the basal lamina of microvessels in the ventral horn of the rabbit spinal cord after transient spinal cord ischemia. Spinal cord sections at the level of L7 were immunostained using monoclonal antibody raised against CB at light and electron microscopic levels. CB immunoreactivity was detected in the basal lamina of microvessels at 30 min after ischemic insult. By 3 h after ischemia, CB immunoreactivity was increased in the basal lamina of the microvessels. CB immunoreactivity began to decrease at 6 h after ischemia and nearly disappeared at 48 h after ischemic insult. For calcium detection in the blood vessels of spinal cord, we conducted an alizarin red staining. Alizarin red reactivity was detected in some microvessels at 3 h after ischemic insult. Our results suggest that the ectopic expression of CB in the microvascular basal laminae may be associated with the buffering of calcium in the endothelial cells of microvessels after ischemic damage. PMID- 15885665 TI - Chronic ether stress-induced response of urocortin 1 neurons in the Edinger Westphal nucleus in the mouse. AB - Urocortin 1 (Ucn1) neurons, most abundantly expressed in the Edinger-Westphal nucleus (E-WN), respond to various acute challenges. In a recent study, we found that acute ether stress resulted in the strongest activation of E-WN Ucn1 cells, as revealed by immunohistochemistry for Fos (often used as a marker for neuronal activation). Although the acute stress responsiveness of E-WN Ucn1 neurons has been widely studied, the activation pattern of Fos in these neurons in response to repeated challenges has not yet been investigated. Therefore, we quantitatively studied Fos activation in E-WN neurons and measured Ucn1 mRNA levels in E-WN neurons after acute and chronic ether stress in mice. Acute stress resulted in a robust Fos response and an increase in Ucn1 mRNA as compared to non stressed mice. In the chronic stress paradigm, Fos expression was unchanged, whereas after 2 and 3 weeks of daily ether exposure Ucn1 mRNA expression had strongly declined in the E-WN. Fos and Ucn1 mRNA were co-expressed in E-WN neurons in both acutely and chronically stressed animals. This paper is the first to demonstrate that Ucn1 mRNA-expressing neurons in the E-WN show a non habituating Fos response to a chronic homotypic ether challenge that also resulted in a reliable down-regulation of E-WN Ucn1 mRNA levels vs. acutely stressed animals. Based on these results, we propose that the E-WN-Ucn1 system represents a novel stress adaptation pathway, which may play an important role in coping with chronic challenges. PMID- 15885666 TI - Properties of neurons in the trigeminal nucleus caudalis responding to noxious dural and facial stimulation. AB - Extracellular single unit recordings were made in the rat trigeminal nucleus caudalis (Vc) from cells with Adelta and C-fibre latency responding to electrical stimulation of the thinned cranium overlying the middle meningeal artery (MMA). The neurons had an ipsilateral facial receptive field (FRF) that mainly extended over areas innervated by the first and second division of the trigeminal nerve but in some cases also included areas innervated by the third division of the trigeminal nerve. No wind-up of either long latency C-fibre or short latency Adelta responses was seen during trains of electrical stimulation. Sensitisation of mechanical stimulation of the FRF could also not be observed at any time during dural stimulation. In contrast, extracellular single unit recordings in the Vc activated by electrical stimulation of the facial skin resulted in a significant wind-up response of long latency response in six of ten cells studied. The facial-elicited wind-up response was significantly enhanced, 18 min after the electrical stimulation protocol was started, indicating that the process of wind-up had generated central excitability. The findings in this study demonstrate a clear difference between the effects of electrical stimulation of cutaneous and non-cutaneous inputs. In the trigeminal system, this has implications for the study of pathways such as those involved in headache, where it is believed that an enhanced dural input to the Vc may generate central sensitisation and partly explain the hyperalgesia and allodynia reported by patients. PMID- 15885667 TI - An electrographic analysis of the synchronous discharge patterns of GEPR-9s generalized seizures. AB - Previous results from our Laboratory have shown a synchronous discharge pattern (less than 1 ms apart) in monopolar recordings from electrodes placed in the cortex, inferior colliculus, and medulla of seizing GEPR-9s. However, the wave morphology of the ictal EEG is quite different for electrodes placed in different anatomical structures. These results lead us to hypothesize that wave morphology was indicative of neural circuitry involved in the GEPR9 seizure and that volume conduction was accounting for synchronous epileptiform EEG pattern. We decided to approach the problem by using a set of two experiments. Experiment 1: Perform a complete precollicular transection in GEPR-9s before inducing seizure in order to observe changes in EEG morphology after forebrain circuitry removal. Experiment 2: A novel methodological approach using a three-dimensional bipolar array enabled the reconstruction of a vector indicative of to which direction is voltage increasing. Such time-varying vector is indicative of the source direction of the high-amplitude epileptiform EEG signal. By placing such an array of electrodes, used to record the 3 bipolar EEGs, in the forebrain, midbrain, and hindbrain, we were able to use a simple intersection method to infer source localization. Our results suggest that the slow wave component of the GEPR9 epileptiform ictal EEG pattern is associated with a midbrain-forebrain circuit while the spike component is associated with a midbrain-hindbrain substrate. These results are supported by experiment 1 in which only the spike component of EEG remained after the precollicular transection. PMID- 15885668 TI - Human adult olfactory neuroepithelial derived progenitors retain telomerase activity and lack apoptotic activity. AB - Olfactory epithelium (OE) contains a population of progenitors responsible for its life-long regenerative capacity. Procedures for the isolation of these progenitors have been established [F.J. Roisen, K.M. Klueber, C.L. Lu, L.M. Hatcher, A. Dozier, C.B. Shields, Adult human olfactory stem cells, Brain Res., 890 (2001) 11-12.] and over 40 patient-specific cell lines from adult postmortem OE and endoscopic biopsy from patients undergoing nasal sinus surgery have been obtained. As these cells emerged in primary cultures, they formed neurospheres (NSFCs). The purpose of the present study was to further characterize these adult human olfactory-derived progenitors. Subcultures of the NSFCs have been passaged nearly 200 times, with a mitotic cycle of 18-20 h. Telomerase activity remains in stem cells; therefore, ELISA was employed to determine the telomerase activity of different lines and passages. Since progenitors undergo low levels of apoptosis, the levels of apoptosis were also examined in these populations. The levels of telomerase and apoptotic activity in 12 NSFC lines remained relatively constant irrespective of donor age, culture duration, or sex. To further study the apoptotic characteristics of the NSFCs, nine different caspases (cysteine proteases) known to be critical in apoptosis were evaluated using gene microarrays comparing cells from a single line at passages 14, 88, and 183. No increases were found in caspase activity in all passages studied. ELISA confirmed the absence of caspase activity over the entire range of passages. This study further suggests that NSFCs can be obtained and used from patients, irrespective of age, sex, or time in culture without altered viability expanding the potential utility of these cells for autologous transplantation and possible diagnostic testing. PMID- 15885669 TI - Synthesis of a glycolipid for studying mechanisms of mitochondrial uncoupling proteins. AB - Glucose-O-omega-palmitic acid is an amphipathic molecule that is useful as a tool for studying the mechanism of mitochondrial uncoupling proteins. The synthesis of this glycolipid is described herein. The study of the reaction of a series of glycosyl donors with appropriate acceptors derived from 16-hydroxyhexadecanoic acid showed that a glycosyl trichloroacetimidate donor was more efficient than thioglycoside, glycosyl halide and glycosyl acetate donors for synthesis of this glycolipid. PMID- 15885670 TI - 1H NMR studies on the hydrogen-bonding network in mono-altro-beta-cyclodextrin and its complex with adamantane-1-carboxylic acid. AB - The hydrogen-bond network in mono-altro-beta-cyclodextrin and in its inclusion complex with adamantane-1-carboxylic acid were investigated by (1)H NMR spectroscopy using the chemical shifts, temperature coefficients and vicinal coupling constants of the exchangeable hydroxy protons. The chemical shifts of the 3-OH signals indicated that the hydrogen-bond network between the 2-OH and 3 OH groups was disturbed not only on each side of the altrose residue, but also along the whole dextrin chain. Upon addition of adamantane-1-carboxylic acid, altrose underwent a conformational change from the (1)C(4) to the (O)S(2) form, allowing a more continuous belt of hydrogen bonding, as evidenced by the downfield shift experienced by the 3-OH proton signals of the glucose residues. PMID- 15885671 TI - Reduced neointima in vein grafts following a blockage of cell recruitment from the vein and the surrounding tissue. AB - OBJECTIVE: Accumulation of intimal smooth muscle cells (SMC) is an important event in vein graft-stenosis. Different SMC sources have been reported, but their interrelations have been poorly studied. In a mouse vein graft model we investigated whether recipient-derived intimal SMCs are recruited from the surrounding tissue and whether blockage of SMC recruitment from the surrounding tissue and/or the donor vein will reduce neointimal formation. METHODS: To detect recipient-derived cells, wild-type veins were implanted into ROSA26 transgenic mice. To block cell recruitment from the surrounding tissue, implanted veins were isolated with a tube-shaped plastic film. To exclude vein-derived cells in the neointimal formation, acellular veins were implanted. RESULTS: In vein grafts isolated from the surrounding tissue the recipient contribution became minimal, but the total number of SMCs was not decreased. Acellular grafts contained an equal number of intimal SMCs as cellular controls after 4 weeks. Isolation of acellular grafts from the surrounding tissue decreased the number of intimal SMCs by 90%. CONCLUSIONS: Recipient-derived SMCs are mainly recruited from the surrounding tissue. Cell recruitment from either the vein or the surrounding tissue is enough to form a neointima. Therefore, a simultaneous inhibition of both these sources is needed to reduce accumulation of intimal SMCs. PMID- 15885672 TI - COX-2 up-regulation and vascular smooth muscle contractile hyperreactivity in spontaneous diabetic db/db mice. AB - OBJECTIVE: Abnormalities in vascular constriction and dilatation are associated with early diabetes and contribute to diabetic vascular complications. However, mechanisms underlying such vascular dysfunction remain to be fully elucidated. The current study tests the role of cyclooxygenase-2 (COX-2) in diabetic vascular smooth muscle dysfunction. METHODS: Small mesenteric artery and aorta are isolated from type 2 diabetic db/db and control mice. Isometric contractions in response to serotonin, angiotensin II, phenylephrine and high potassium are determined in small spiral mesenteric arterial or aortic strips. COX-2 mRNA and protein levels are analyzed by using DNA microarray, real-time PCR and immunoblot. RESULTS: Contractions induced by serotonin, angiotensin II, phenylephrine and high potassium are significantly higher in endothelium-denuded smooth muscle strips isolated from db/db mice than in those isolated from control mice. The contractile hyperreactivity is observed in aortic and third-order branch small mesenteric arterial smooth muscle strips. DNA microarray, real-time PCR and immunoblot analysis show that compared with control mice, COX-2 mRNA and protein are significantly increased in db/db mice aortic smooth muscle. The COX-2 up-regulation is temporally associated with the development of diabetes mellitus and vascular smooth muscle contractile hyperreactivity. Inhibition of COX-2 with NS-398 or SC-58125 partially--but significantly--alleviates agonist-induced but not potassium-induced contractile hyperreactivity. In addition, serum isolated from db/db mice induces COX-2 expression and increases thromboxane A2 production in primary cultured vascular smooth muscle cells (VSMC). SQ-29548, a TP receptor antagonist, diminished the db/db mice vascular smooth muscle contractile hyperreactivity. CONCLUSIONS: COX-2 is up-regulated and contributes at least in part to the vascular smooth muscle contractile hyperreactivity in db/db mice. PMID- 15885673 TI - PDGF-induced signaling in proliferating and differentiated vascular smooth muscle: effects of altered intracellular Ca2+ regulation. AB - OBJECTIVE: Platelet-derived growth factor-BB (PDGF)-induced intracellular signaling is involved in phenotypic modulation of vascular smooth muscle (VSM). This study has examined the PDGF-induced Ca2+ increase and the resultant effect on signaling pathways in proliferative compared with fully differentiated VSM. METHODS: PDGF-induced changes in Ca2+ were measured in portal vein (PV) myocytes from 2-4-day-old (proliferating), compared to 6-week-old (differentiated), Sprague Dawley rats. Phospholipase C (PLC)gamma expression and activation of extracellular signal-regulated kinase (ERK) 1/2 was determined by immunoblotting or confocal immunolabelling. Activation of the Ca(2+)-dependent transcription factor, nuclear factor of activated T-cells (NFATc), was assessed by electromobility shift assay. RESULTS: PDGF increased the intracellular Ca2+ concentration in differentiated, but not in proliferating, PV myocytes. This is probably due to very low expression of PLC(gamma) in proliferating PV. In 6-week old PV, PDGF stimulation induced nuclear translocation and activation of NFATc. PDGF did not induce NFATc activation in neonatal PV. PDGF-induced ERK1/2 activation was observed in both 2-4-day-old and 6-week-old PV. In 6-week-old PV, ERK1/2 activation was Ca(2+)-dependent and protein kinase C-dependent. However in 2-4-day-old PV, PDGF-induced ERK1/2 activation was via a Ca(2+)-independent, atypical protein kinase C. PLC(gamma) expression was also decreased in the neointima, compared to media, of balloon-injured rabbit subclavian arteries. CONCLUSIONS: The regulation of PDGF-induced Ca2+ increases by PLC(gamma) expression in VSM may provide a mechanism for coordinating different signaling pathways leading to activation of specific transcription factors. This may play an important role in the phenotypic modulation of VSM. PMID- 15885674 TI - Abnormal conduction and repolarization in late-activated myocardium of dyssynchronously contracting hearts. AB - BACKGROUND: Cardiac dyssynchrony due to intraventricular conduction delay produces heterogeneous regional wall stress and worsens arrhythmia susceptibility in failing hearts. We examined whether chronic dyssynchrony per se induces regionally heterogeneous electrophysiological remodeling. METHODS AND RESULTS: Adult dogs (n=9) underwent left bundle branch radiofrequency ablation (QRS duration increased from 50+/-7 to 104+/-7 ms); 6 untreated dogs served as controls. A subset of ablated (n=3) and control (n=4) dogs underwent tagged MR imaging to confirm ablation-induced left ventricular (LV) dyssynchrony. Four weeks later, hearts were excised and early (anterior)- and late (lateral) activated myocardial segments were isolated. Conduction velocity (CV), action potential duration (APD), and refractory period (RP) of paced, arterially perfused myocardial wedges were studied by extracellular and optical mapping, and arrhythmia susceptibility was assessed by programmed stimulation. Regional stress response kinase, calcium cycling, and gap junction protein expression were assayed by Western blotting, and the subcellular distribution of connexin43 was analyzed by immunofluorescence microscopy. CV, APD, and RP were significantly reduced in the late-activated, lateral wall of dyssynchronous hearts compared to the anterior wall. Normal differences in CV (endocardial>epicardial) were reversed in the dyssynchronous lateral LV. While the total expression of connexin43 was unaltered in dyssynchronous models, its subcellular location was redistributed in late-activated myocardium from intercalated discs to lateral myocyte membranes. Arrhythmias were rare in tissue from normal and dyssynchronous models. Total expression of calcium-cycling proteins (sarcoplasmic reticulum Ca2+ ATPase and phospholamban) and the stress-response kinase phospho-ERK did not vary regionally in either model. CONCLUSIONS: Dyssynchrony even in the absence of LV dysfunction induces regionally specific changes in conduction and repolarization. These changes support a novel mechanism linking mechanical dyssynchrony to persistent electrophysiological remodeling and heterogeneity. PMID- 15885675 TI - Role of mitochondrial re-energization and Ca2+ influx in reperfusion injury of metabolically inhibited cardiac myocytes. AB - OBJECTIVE: We used isolated myocytes to investigate the role of mitochondrial re energization and Ca2+ influx during reperfusion on hypercontracture, loss of Ca2+ homeostasis and contractile function. METHODS: Isolated adult rat ventricular myocytes were exposed to metabolic inhibition (NaCN and iodoacetate) and reperfusion injury was assessed from hypercontracture, loss of Ca2+ homeostasis ([Ca2+]i measured with fura-2) and failure of contraction in response to electrical stimulation. Mitochondrial membrane potential was followed using the potentiometric dye tetramethylrhodamine ethyl ester. RESULTS: Metabolic inhibition led to contractile failure and rigor accompanied by a sustained increase in [Ca2+]i. Reperfusion after 10 min metabolic inhibition led to an abrupt repolarization of the mitochondrial membrane potential (after 25.5+/-1.2 s), a transient fall in [Ca(2+]i followed by an abrupt hypercontracture (37.1+/ 1.8 s) in 84% of myocytes. Ca2+ homeostasis (diastolic [Ca2+]i < 250 nM) recovered in only 23.3+/-5.1% of cells and contractions recovered in 15.3+/-2.2%. Oligomycin abolished the hypercontracture on reperfusion, but mitochondrial repolarization was unaffected. Preventing Ca2+ influx during reperfusion with [Ca2+]i-free Tyrode or with an inhibitor of Na(+)/Ca2+ exchange did not prevent the hypercontracture, but increased the percentage of cells recovering Ca2+ homeostasis and contractile function. The presence of 0.5 microM cyclosporin A did not prevent hypercontracture but increased the percentage of cells recovering Ca2+ homeostasis to 56.2+/-3.6% and contractile function to 52+/-4.3%. CONCLUSIONS: Reperfusion-induced hypercontracture, and loss of Ca2+ homeostasis and contractile function are initiated following mitochondrial re-energization. The hypercontracture requires the production of oxidative ATP but not Ca2+ influx during reperfusion. Loss of Ca2+ homeostasis and contractile function are linked to Ca2+ influx during reperfusion, probably via opening of mitochondrial permeability transition pores. PMID- 15885676 TI - Catechins prevent vascular smooth muscle cell invasion by inhibiting MT1-MMP activity and MMP-2 expression. AB - OBJECTIVE: Regular consumption of green tea is associated with a reduced risk of mortality due to coronary diseases and cancer. The present study examined whether a green tea extract (GTE) inhibits activation of matrix metalloproteinase-2 (MMP 2), a major collagenase involved in vascular remodeling of atherosclerotic plaques, in vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: The expression of MMP-2 was assessed by Northern and Western blot analyses in human aortic VSMCs. MMP-2 activity was evaluated by zymography, membrane-type1-MMP (MT1 MMP, MMP-14) activity by an enzymatic assay, and cell invasion by a modified Boyden chamber assay. The thrombin-induced activation of secreted MMP-2 was abolished by GTE and the green tea polyphenols (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG). GTE reduced the expression of MMP-2 mRNA and protein. GTE, EGCG and ECG directly inhibited cell-associated MT1-MMP activity, the physiological activator of MMP-2, in a reversible manner. Thrombin stimulated VSMCs invasion was abolished by EGCG and ECG, and reduced by GTE. CONCLUSIONS: GTE inhibits thrombin-induced VSMCs invasion most likely by preventing MMP-2 expression and its activation by a direct inhibition of MT1-MMP. The ability of green tea to prevent cell invasion and matrix degradation might contribute to its protective effect on atherosclerosis and cancer. PMID- 15885677 TI - Immunostimulatory activity of major membrane protein-II from Mycobacterium leprae. AB - We examined the antigenicity of an immunomodulatory protein, major membrane protein (MMP)-II, from Mycobacterium leprae, since host defense against M. leprae largely depends on adaptive immunity. Both unprimed and memory T cells from healthy individuals were stimulated by autologous MMP-II-pulsed monocyte-derived dendritic cells (DCs) to produce IFN-gamma. The DC-mediated IFN-gamma production was dependent on the expression of MHC, CD86, and MMP-II antigens. Memory T cells from paucibacillary (PB) leprosy more extensively responded to MMP-II-pulsed DCs than T cells from healthy individuals, while comparable IFN-gamma was produced by unprimed T cells. Memory T cells from multibacillary leprosy, which are normally believed to be anergic, were activated similarly to those from healthy individuals by MMP-II-pulsed DCs. These results suggest that memory T cells from PB leprosy are primed with MMP-II prior to the manifestation of the disease, and MMP-II is highly antigenic in terms of activation of adaptive immunity. PMID- 15885678 TI - Cytochrome P450 enzyme-mediated degradation of Echinacea alkylamides in human liver microsomes. AB - Echinacea preparations are widely used herbal remedies for the prevention and treatment of colds. In this study we have investigated the metabolism by human liver microsomes of the alkylamide components from an Echinacea preparation as well as that of pure synthetic alkylamides. No significant degradation of alkylamides was evident in cytosolic fractions. Time- and NADPH-dependent degradation of alkylamides was observed in microsomal fractions suggesting they are metabolised by cytochrome P450 (P450) enzymes in human liver. There was a difference in the susceptibility of 2-ene and 2,4-diene pure synthetic alkylamides to microsomal degradation with (2E)-N-isobutylundeca-2-ene-8,10 diynamide (1) metabolised to only a tenth the extent of (2E,4E,8Z,10Z)-N isobutyldodeca-2,4,8,10-tetraenamide (3) under identical incubation conditions. Markedly less degradation of 3 was evident in the mixture of alkylamides present in an ethanolic Echinacea extract, suggesting that metabolism by liver P450s was dependent both on their chemistry and the combination present in the incubation. Co-incubation of 1 with 3 at equimolar concentrations resulted in a significant decrease in the metabolism of 3 by liver microsomes. This inhibition by 1, which has a terminal alkyne moiety, was found to be time- and concentration-dependent, and due to a mechanism-based inactivation of the P450s. Alkylamide metabolites were detected and found to be the predicted epoxidation, hydroxylation and dealkylation products. These findings suggest that Echinacea may effect the P450 mediated metabolism of other concurrently ingested pharmaceuticals. PMID- 15885679 TI - Non-Hodgkin lymphoma and benzene exposure: a systematic literature review. AB - Epidemiological papers that address an association between benzene exposure and non-Hodgkin lymphoma (NHL) were identified and separated by study design. Eighteen studies contained 21 study groups-11 population-based case-control study groups, 3 nested occupation-based case-control study groups and 7 occupational benzene cohort study groups. Petroleum industry studies were not included. Only two of these 21 study groups showed statistically significant associations. However, these were among workers with multiple exposures across industries. Eleven of the 21 study groups presented ratios less than one, two equaled one, and eight were greater than one. Over all, about as many cases were observed (404) as expected (390.0) for an observed to expected ratio of 1.04 (0.94-1.14). After removal of the studies with multiple chemical exposures problems, the observed was 359 cases with 373.2 cases expected, yielding an odds ratio of 0.96 (0.86-1.06). Further assessment of an association with NHL should document the benzene exposure and separate out the contribution of non-benzene exposures. PMID- 15885680 TI - Hypochlorous acid-induced stress on human spermatozoa. A model for inflammation in the male genital tract. AB - The fertilising ability of human spermatozoa may be impaired by inflammations of the genital tract, although details of these processes are still unknown. Hypochlorous acid (HOCl), an important product of myeloperoxidase released from stimulated neutrophils, induces a concentration-dependent increase in externalisation of phosphatidylserine in ejaculated human spermatozoa as revealed by fluorescence-activated cell sorting (FACS) analysis. The increase of annexin-V binding cells starts already at about 10(-5) mol/l HOCl, while a formation of lysophosphatidylcholines as detected by matrix-assisted laser desorption and ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is only found at HOCl concentrations higher than 10(-4) mol/l. Thus, changes in lipid composition of spermatozoa are unlikely responsible for the phosphatidylcholine (PS) externalisation. These data gave concomitant evidence that HOCl itself leads to a dramatic damage of the cell membrane. Thus, the neutrophil-derived HOCl contributes to the deterioration of spermatozoa leading to diminished fertilisation ability. PMID- 15885681 TI - EDTA should be the anticoagulant of choice for the measurement of plasma ammonia: report of a problem sample. PMID- 15885682 TI - Iron overload. AB - Iron overload disorders represent a heterogenous group of conditions resulting from inherited and acquired causes. With the discovery of new proteins and genetic defects we have gained greater insight into their causation at the molecular level and the complex mechanisms of normal and disordered iron homeostasis. Here we review the normal mechanisms and regulation of gastrointestinal iron absorption and liver iron transport and their dysregulation in iron overload states. Advances in the understanding of the natural history of iron overload disorders and new methods for clinical detection and management of hereditary hemochromatosis are also reviewed. PMID- 15885683 TI - Pesticide-induced alteration in mice hepato-oxidative status and protective effects of black tea extract. AB - BACKGROUND: This study investigates the role of black tea extract in protection against oxidative damage induced in mice by a subacute oral dose of a combination of pesticides. METHODS: Liver damage markers like amino-acid transferases and alkaline phosphatase, oxidative damage markers indicating extent of lipid peroxidation, host antioxidant reserve indicators like reduced glutathione (GSH) and total thiol levels, GSH-dependent enzyme activities viz., glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR) superoxide dismutase (SOD) and catalase (CAT) were measured by spectrophotometric techniques in four different groups namely (1) control, (2) black tea extract, (3) pesticide alone and (4) pretreatment with black tea extract followed by pesticide treatment. RESULTS: Following exposure to pesticides lipid peroxidation was increased compared to that in the control [0.50+/-0.083 (mean+/-S.E.) vs. 0.21+/-0.02 micromol/mg protein, p<0.001]. Moreover, levels of antioxidants like GSH and total thiol were also significantly decreased in comparison to control, e.g., GSH [5.16+/-0.78 vs. 6.96+/-1.35 micromol/mg protein, p<0.001], total thiol [19.3+/-2.74 vs. 26.15+/-2.8 micromol/mg protein, p<0.001]. In addition, the activities of antioxidant enzymes like GPx, GST, GR, SOD and catalase were also likewise diminished by oxidant damage. Treatment with aqueous black tea extract significantly protects the liver tissue from the oxidative damage and shifts the trend towards amelioration and replenishment of the antioxidant status. CONCLUSIONS: The active components present in natural extracts like that of black tea can be very effective in perhaps reducing the extent of injury and in overcoming oxidant damage caused by exposure to environmental agents like pesticides. PMID- 15885684 TI - Principal component analysis of some oxidative stress parameters and their relationships in hemodialytic and transplanted patients. AB - BACKGROUND: Oxidative stress profoundly influences the biochemistry of proteins and many other molecules in tissues of uremic patients. In three different groups of uremic patients, the concentrations of the free and bound pentosidine and low molecular-weight-advanced glycoxydation end products (LMW-AGEs), carbonyls (LMW C), advanced oxidation protein products (AOPP) and the total antioxidant power of serum were studied in order to determine the relationships between these factors in hemodialytic and transplanted patients. PATIENTS AND METHODS: The above mentioned parameters were determined in 10 subjects who were currently in hemodialysis (HD) treatment, 10 kidney transplanted patients with chronic renal failure (Tx-CRF), 10 kidney transplanted patients with normal renal function (Tx N) and 10 healthy subjects (Ctr). The data matrix (40x7) was analyzed using the principal component analysis (PCA). RESULTS: AGEs, carbonyls and AOPP were strongly correlated, while the total antioxidative serum capacity was not related to the other oxidative stress parameters. All the oxidative stress-related parameter values (AGEs, AOPP and LMW-C) in the Tx patients were similar to those of the control group, but were higher in the patients with chronic renal failure. CONCLUSIONS: The correlation between early and advanced oxidative stress markers indicates that reactive oxygen species are involved in a common step in the mechanism of protein modification in all the patient examined. The relationships between carbonyls and AGEs (free, bound pentosidine and LMW-AGEs) support the hypothesis of "carbonyl stress". The common mechanism of the formation of oxidation products in healthy and diseased subject suggests their role of detoxification within kidney function. The total antioxidant power of the serum is not related to the other parameters, which indicates a possible role of molecule interfering. PMID- 15885685 TI - Pitfalls of immunoassay and sample for IGF-I: comparison of different assay methodologies using various fresh and stored serum samples. AB - OBJECTIVE: Determination of insulin-like growth factor (IGF)-I is now a routine adjunct to multiple research and clinical investigations. Evidence has associated higher IGF-I levels with various human pathologies, but the reported associations have not been invariably confirmed. We examined the potential for post-sampling proteolysis and evaluated the impact of such events on IGF-I immunoassays. DESIGN AND METHODS: We compared IGF-I in different sets of fresh and frozen old samples using four different and commonly used immunoassays. The potential for post sampling proteolysis was further examined by assaying fresh samples stored for 4 weeks at various temperatures in the absence or presence of protease inhibitors. RESULTS: IGF-I levels in fresh serum samples from adult males, females, and pregnant subjects by all methods were similar and were highly correlated (r=0.85 0.97). The same was true for levels in frozen ( approximately 2 years at --80 degrees C) samples from diabetic patients, which are reportedly associated with enhanced proteolytic activity. In contrast, in another set of frozen adult male and female samples ( approximately 8 years at --20 degrees C), the inter-method median IGF-I levels varied by approximately 3- to 4-fold and the values poorly correlated. Similar variability in the inter-method response was also observed when IGF-I in the replicates of fresh samples stored at 4 degrees C for 4 weeks was measured. However, the 4 degrees C storage effect could be completely blocked by the addition of protease inhibitors, allowing for all assays to detect 92- 101% of the expected mean levels. CONCLUSIONS: The data indicate susceptibility of IGF-I to significant post-sampling proteolysis and suggest the importance of immunoassays for the intact molecule. Immunoassays that lack specificity for intact IGF-I may mask the potential pathophysiological effects of proteolysis and generate misleading results, particularly in studies involving inappropriately stored and/or proteolyzed samples. In such cases, underestimation of the in vivo levels by the intact assays would occur, but the findings of low IGF-I levels may be indicative of questionable sample quality. PMID- 15885686 TI - TRIM37 defective in mulibrey nanism is a novel RING finger ubiquitin E3 ligase. AB - Mulibrey nanism is an autosomal recessive prenatal-onset growth disorder characterized by dysmorphic features, cardiomyopathy, and hepatomegaly. Mutations in TRIM37 encoding a tripartite motif (TRIM, RING-B-box-coiled-coil)-family protein underlie mulibrey nanism. We investigated the ubiquitin ligase activity predicted for the RING domain of TRIM37 by analyzing its autoubiquitination. Full length TRIM37 and its TRIM domain were highly polyubiquitinated when co-expressed with ubiquitin. Polyubiquitination was decreased in a mutant protein with disrupted RING domain (Cys35Ser;Cys36Ser) and in the Leu76Pro mutant protein, a disease-associated missense mutation affecting the TRIM domain of TRIM37. Bacterially produced GST-TRIM domain fusion protein, but not its Cys35Ser;Cys36Ser or Leu76Pro mutants, were polyubiquitinated in cell-free conditions, implying RING-dependent modification. Ubiquitin was also identified as an interaction partner for TRIM37 in a yeast two-hybrid screen. Ectopically expressed TRIM37 rapidly formed aggregates that were ubiquitin-, proteasome subunit-, and chaperone-positive in immunofluorescence analysis, defining them as aggresomes. The Cys35Ser;Cys36Ser mutant and the Leu76Pro and Gly322Val patient mutant proteins were markedly less prone to aggregation, implying that aggresomal targeting reflects a physiological function of TRIM37. These findings suggest that TRIM37 acts as a TRIM domain-dependent E3 ubiquitin ligase and imply defective ubiquitin-dependent degradation of an as-yet-unidentified target protein in the pathogenesis of mulibrey nanism. PMID- 15885687 TI - Toxocara canis: impact of preweaning nutritional deprivation on the pathogenesis of pneumonia in the mouse. AB - The present study was conducted to better understand the immure response to Toxocara canis pneumonia in mice with preweaning nutritional deprivation. Breast fed Swiss mice, undernourished due to large litter size (up to 15 pups) and paired controls with only 5-8 pups were used. At 21 days old, both groups were infected with T. canis larvae. Liver retinol, retinyl palmitate, and inflammatory infiltrate in lungs were compared in both groups. Significantly lower levels of retinol and retinyl palmitate in liver tissue confirmed the hypovitaminosis A (P < 0.0001 for both comparisons) in the nutritionally deprived animals. Histological analysis showed similar eosinophilic infiltration in both groups at day 3 but was significantly more severe in undernourished mice at day 20 post infection (P = 0.01). The present findings indicate that preweaning undernourishment is associated with a more severe inflammation in response to T. canis pneumonia. It suggests that vitamin A deficiency that persists after nutritional rehabilitation, may contribute to the severity of T. canis infection. The authors suggest that nutritional status should be carefully investigated in patients with more severe clinical findings. PMID- 15885688 TI - Simultaneous spectrofluorimetric and spectrophotometric determination of melatonin and pyridoxine in pharmaceutical preparations by multivariate calibration methods. AB - Partial least-squares (PLS) calibration and principal component regression (PCR) methods were utilized for the simultaneous spectrofluorimetric and spectrophotometric determination of pyridoxine (PY) and melatonin (MT). Since emission and adsorption spectra of these drugs overlap, PY and MT cannot be directly determined by fluorimetric nor by spectrophotometric methods. Full spectrum multivariate calibration PLS and PCR methods were developed for both fluorimetry and spectrophotometry. The conditions were optimized for fluorimetric as well as for spectrophotometric determination of both drugs. The simultaneous determination of PY and MT was carried out in mixtures by recording the emission fluorescence spectrum between 324 and 500 nm (lambda(ex) 285 nm) for fluorimetry, and by recording the absorption spectrum between 250 and 350 nm for spectrophotometry (lambda(max(PY)) 310 nm, lambda(max(MT)) 278 nm). The experimental calibration matrixes were designed orthogonally. At the optimum conditions, dynamic ranges were 0.04-1.3 and 0.1-4 microg ml(-1) for fluorimetry and 1-22 and 1-24 microg ml(-1) for spectrophotometry for MT and PY, respectively. The calibration concentrations were prepared in the dynamic ranges. The parameters of the chemometrics procedure for the simultaneous determination of MT and PY were optimized, and the proposed methods were validated with prediction set. Finally the procedures were successfully applied to simultaneous spectrofluorimetric and spectrophotometric determination of PY and MT in synthetic mixtures and in a pharmaceutical formulation. PMID- 15885689 TI - Synthesis and biological evaluation of pro-drugs of GW196771, a potent glycine antagonist acting at the NMDA receptor. AB - GW196771 is a potent antagonist of the modulatory glycine site of the N-methyl-D aspartate (NMDA) receptor exhibiting outstanding in vivo profile in different animal models of chronic pain. With the aim to maximize the drug delivery to the target organs a suitable "pro-drug approach" was attempted; in this regards two conjugates of GW196771 with nutrients actively transported into the brain, namely adenosine and glucose, were prepared and investigated. These compounds, were evaluated in vitro in terms of their stability in rat plasma and in vivo on rats. Although an improvement was observed in terms of brain penetration of the esters vs. the parent compound, the amount of the latter did not increase significantly, probably due to some degradation events in the brain, different from the expected ester hydrolysis, resulting in a reduced availability of GW196771. PMID- 15885691 TI - Distinctive stress effects on learning during puberty. AB - Puberty is a time of significant change in preparation for adulthood. Here, we examined how stressful experience affects cognitive and related hormonal responses in male and female rats prior to, during and after puberty. Groups were exposed to an acute stressor of brief periodic tailshocks and tested 24 h later in an associative memory task of trace eyeblink conditioning. Exposure to the stressor did not alter conditioning in males or females prior to puberty but enhanced conditioning in both males and females during puberty. The enhancement occurred in pubescent females irrespective of the estrous cycle. In adulthood, sex differences in trace conditioning and the response to stress emerged: females outperformed males under unstressed conditions, but after stressor exposure, trace conditioning in females was impaired whereas that in males was enhanced. These differences were not related to changes in gross motor activity or other nonspecific measures of performance. The effects of acute stress on corticosterone, estradiol, progesterone, and testosterone were also measured. Stressor exposure increased the concentration of corticosterone in all age groups, although sex differences were only evident in adults. All reproductive hormones except estradiol increased with age in a predictable and sex dependent fashion and none were affected by stressor exposure. Estradiol decreased in male rats across age, and remained stable for female rats. Together, these data indicate that males and female respond similarly to learning opportunities and stressful experience before and during puberty; it is in adulthood that sex differences and the opposite responses to stress arise. PMID- 15885690 TI - The vertebrate social behavior network: evolutionary themes and variations. AB - Based on a wide variety of data, it is now clear that birds and teleost (bony) fish possess a core "social behavior network" within the basal forebrain and midbrain that is homologous to the social behavior network of mammals. The nodes of this network are reciprocally connected, contain receptors for sex steroid hormones, and are involved in multiple forms of social behavior. Other hodological features and neuropeptide distributions are likewise very similar across taxa. This evolutionary conservation represents a boon for experiments on phenotypic behavioral variation, as the extraordinary social diversity of teleost fish and songbirds can now be used to generate broadly relevant insights into issues of brain function that are not particularly tractable in other vertebrate groups. Two such lines of research are presented here, each of which addresses functional variation within the network as it relates to divergent patterns of social behavior. In the first set of experiments, we have used a sexually polymorphic fish to demonstrate that natural selection can operate independently on hypothalamic neuroendocrine functions that are relevant for (1) gonadal regulation and (2) sex-typical behavioral modulation. In the second set of experiments, we have exploited the diversity of avian social organizations and ecologies to isolate species-typical group size as a quasi-independent variable. These experiments have shown that specific areas and peptidergic components of the social behavior network possess functional properties that evolve in parallel with divergence and convergence in sociality. PMID- 15885692 TI - New graduate nurse self-concept and retention: a longitudinal survey. AB - Graduate nurse attrition is an increasing phenomenon within a world of decreasing nursing numbers. The newly developed nursing self-concept of the graduate nurse may provide a key indicator for predicting graduate retention. This study explores the development of multiple dimensions of nursing self-concept and examines their relationship to graduate nurse retention plans. A descriptive survey design with repeated measures was utilised to assess nurse self-concept and retention plans. The key findings suggest that multiple dimensions of graduate nurse self-concepts rise significantly in the second half of their graduate year and that nurse general self-concept is a strong predictor of graduate nurse retention. The implications of this study are that monitoring of self-concept throughout the transitional period for new nurses can lead to early detection and appropriate intervention strategies thereby improving retention rates for new nurses. PMID- 15885693 TI - Multi-host parasite species in cophylogenetic studies. AB - Cophylogenetic studies examine the relationship between host and parasite evolution. One aspect of cophylogenetic studies that has had little modern discussion is parasites with multiple definitive hosts. Parasite species with multiple host species are anomalous as, under a codivergence paradigm, speciation by the hosts should cause speciation of their parasites. We discuss situations such as cryptic parasite species, recent host switching or failure to speciate that may generate multi-host parasites. We suggest methods to identify which of the mechanisms have led to multi-host parasitism. Applying the suggested methods may allow multi-host parasites to be integrated more fully into cophylogenetic studies. PMID- 15885694 TI - The 3A1-La monoclonal antibody reveals key features of Leishmania (L) amazonensis metacyclic promastigotes and inhibits procyclics attachment to the sand fly midgut. AB - In this work, we characterise metacyclic promastigotes of Leishmania amazonensis, the causative agent of cutaneous and diffuse cutaneous leishmaniasis in the New World. To purify metacyclics from stationary culture by negative selection, we used the monoclonal antibody 3A1-La produced against procyclic promastigotes. The purified forms named 3A1-La(-) promastigotes, present key metacyclic characteristics: slender cell body and long flagella, ultrastructural features, resistance to complement lysis, high infectivity for macrophages and mice and reduced capacity for binding to the sand fly midgut. Moreover, the epitope recognised by 3A1-La is important for the promastigote attachment to the insect vector midgut epithelium. These results further characterise 3A1-La(-) promastigotes as metacyclic forms of L. amazonensis. PMID- 15885695 TI - Reduction in transplacental transmission of Neospora caninum in outbred mice by vaccination. AB - Infection with the protozoan parasite Neospora caninum is an important cause of abortion in cattle. A major source of infection is transplacental transfer of the parasite from mother to offspring during pregnancy. This study describes investigations on the immunisation of outbred Qs mice before pregnancy with live or a crude lysate of N. caninum (NC-Nowra isolate) to prevent transplacental transfer of a challenge infection administered during pregnancy. Parasites present in the brains of pups from mice challenged with N. caninum (NC-Liverpool) were detected by PCR. Injection of live NC-Nowra tachyzoites before pregnancy dramatically reduced transplacental transfer from 75 to 0.8% in one experiment and from 76 to 8% in a second experiment. Injection of a crude lysate of NC-Nowra tachyzoites reduced transplacental transfer from 67 to 53% in one experiment and from 76 to 63% in a second experiment. Analysis of N. caninum-specific IgG1 and IgG2a antibody levels prior to pregnancy and challenge showed that NC-Nowra lysate induced a response skewed towards IgG1 whereas live parasites induced both IgG1 and IgG2a antibodies. After pregnancy and a challenge infection, a similar IgG1/IgG2a response was seen in all challenged groups. These results provide further positive support for the hypothesis that transplacental transmission of this parasite is preventable by vaccination. PMID- 15885696 TI - Development of the egg hatch assay for detection of anthelminthic resistance in human hookworms. AB - Evidence of development and rapid spread of anthelminthic resistance in veterinary nematodes raises concern that the increasingly frequent treatments used in chemotherapy-based programmes to control human soil-transmitted helminths may select resistant worms. The aim of this study was to adapt, refine, and evaluate the Egg Hatch Assay (EHA) test, which has been used for veterinary nematodes, for field testing of benzimidazole (BZ) susceptibility/resistance in human hookworms. A second objective was to use this EHA to assess whether a population of worms resistant to mebendazole (MBZ) has built up in a sub population of frequently treated children in Pemba Island. Stools from 470 school children enrolled in the first (Standard 1) and in the fifth (Standard 5) class were examined at baseline and at 21 days after treatment with 500 mg MBZ or placebo tablets. Standard 1 children had never received any MBZ treatment whilst Standard 5 children had received a total of 13 rounds of treatment. The EHA, involving culture of purified eggs with increasing drug concentrations showed that, for thiabendazole (TBZ), the mean ED(50)s (concentrations required to prevent 50% of the viable eggs from hatching) for all children at baseline were 0.079 microg/ml at 48h and 0.120 microg/ml at 72h (P<0.001). For MBZ, the mean ED(50)s for all children at baseline were 0.895 microg/ml at 48h and 1.50 microg/ml at 72h (P<0.001). For TBZ and for MBZ the ED(50) from Standard 1 were similar to those from Standard 5 children both at 48 and at 72h. At the follow-up for TBZ and for MBZ, there was no significant difference between the ED(50) from children who had received MBZ and children treated with placebo. In Pemba, TBZ ED(50) values of children non-exposed (Standard 1) and of children exposed (Standard 5) to MBZ treatment, and data from children treated with MBZ and placebo indicate that a drug-resistant worm population has not built up within treated individuals, and that periodic treatment has not yet selected for widespread BZ resistance, at least at the threshold detectable by the EHA in this study. However, ED(50) values for strains isolated from Mafia island, an area never exposed to BZ treatment were lower than for Pemba, suggesting lowered sensitivity of hookworm eggs recovered from Pembian children towards BZ. PMID- 15885697 TI - Searching new targets for anthelminthic strategies: Interference with glycosphingolipid biosynthesis and phosphorylcholine metabolism affects development of Caenorhabditis elegans. AB - Nematode infections are amongst the most abundant diseases of man and animals. They are characterised by a low mortality but high morbidity, thus reflecting the adaptation of these parasites to their hosts. Resistance as well as severe side effects and efficacies restricted to distinct larval stages or parasites of the anthelmithics used at present require the urgent development of new and more nematode-specific drugs, targeting enzymes of parasite restricted biosynthetic routes. Caenorhabditis elegans has been found to be a good model system for parasitic nematodes, drug screening and developmental studies. Structural analyses have revealed nematode-specific glycosphingolipid structures of the arthro-series, carrying in part, phosphorylcholine substituents. These biomolecules appear to play important roles in nematode development, fertility and survival within the host and are, therefore, good target-candidates for the development of new anthelminthic strategies. Here we show that RNAi experiments targeting enzymes of glycosphingolipid biosynthesis or choline metabolism result, in part, in a drastic reduction of fertility. We further tested various chemical inhibitors of these pathways and found significant effects on the development of the worms, resulting in developmental arrest, sterility and, in part, lethality. Such inhibitors can, therefore, help to define new classes of anthelminthics. PMID- 15885698 TI - Design and implementation of an instrumented ulnar head prosthesis to measure loads in vitro. AB - The development of a novel instrumented implant for ulnar head replacement is presented in this study. This implant was instrumented with strain gauges to quantify bending moments about the anatomic axes of the distal ulna, and subsequently the distal radioulnar joint (DRUJ) reaction force magnitude. The implant was surgically inserted in seven cadaveric upper extremities, which were subsequently mounted in a custom joint simulator. Simulated active unresisted pronation and supination motion trials were conducted using computer-controlled pneumatic actuators to simulate forearm musculature. Passive (unloaded) trials were also conducted. The reaction force across the DRUJ ranged from 2 to 10 N in magnitude during this unresisted motion. Increased bending moment magnitudes were measured when the forearm was positioned in supination compared to pronation. The magnitude of joint bending moments showed a consistent pattern with forearm position, regardless of simulated active or passive rotation, or supination and pronation motion trials. This result illustrates that the primary influence on joint load is likely the position and contact with the radial articulation. This study of DRUJ loading should be useful for biomechanical modeling, implant design considerations and improved knowledge of articular mechanics. PMID- 15885699 TI - The effects of aneurysm on the biaxial mechanical behavior of human abdominal aorta. AB - The biomechanical response of normal and pathologic human abdominal aortic tissue to uniaxial loading conditions is insufficient for the characterization of its three-dimensional (3D) mechanical behavior. Planar biaxial mechanical evaluation allows for 3D constitutive modeling of nearly incompressible tissues, as well as the investigation of the nature of mechanical anisotropy. In the current study, 26 abdominal aortic aneurysm (AAA) tissue samples and 8 age-matched (> 60 years of age) nonaneurysmal abdominal aortic (AA) tissue samples were obtained and tested using a tension-controlled biaxial testing protocol. Graphical response functions (Sun et al., 2003. J. Biomech. Eng. 125, 372-380) were used as a guide to describe the pseudo-elastic response of AA and AAA. Based on the observed pseudo-elastic response, a four-parameter exponential strain energy function developed by Vito (1990. J. Biomech. Eng. 112, 153-159) was used from which both an individual specimen and group material parameter sets were determined for both AA and AAA. Peak Green strain values in the circumferential (Ethetatheta,max) and longitudinal (ELL,max) directions under an equibiaxial tension of 120 N/m were also compared. The strain energy function fit all of the individual specimens well with an average R2 of 0.95 +/- 0.02 and 0.90 +/- 0.02 (mean +/- SEM) for the AA and AAA groups, respectively. The average Ethetatheta,max at 200 N/m equibiaxial tension was found to be significantly smaller for AAAs as compared to AAs (0.07 +/- 0.01 versus 0.13 +/- 0.03, respectively; p < 0.01). There was also a pronounced increase in the circumferential stiffness for AAA tissue as compared to AA tissue, indicating a larger degree of anisotropy for this tissue as compared to age-matched AA tissue. We also observed that the four-parameter Fung elastic model was not able to fit the AAA tissue mechanical response using physically realistic material parameter values. It was concluded that aneurysmal degeneration of the abdominal aorta is associated with an increase in mechanical anisotropy, with preferential stiffening in the circumferential direction. PMID- 15885700 TI - Reproductive division of labor, dominance, and ecdysteroid levels in hemolymph and ovary of the bumble bee Bombus terrestris. AB - To determine whether ecdysteroids are associated with reproductive division of labor in Bombus terrestris, we measured their levels in hemolymph and ovaries of queens and workers. Queens heading colonies had large active ovaries with high ecdysteroid content, whereas virgin gynes and mated queens before and after diapause had undeveloped ovaries with low ecdysteroid content. The hemolymph ecdysteroid titer was rather variable, but in a pooled analysis of mated queens before and after diapause versus colony-heading queens, ecdysteroid titers were higher in the latter group. In workers, agonistic behavior, ovarian activity, ovarian ecdysteroid content, and hemolymph ecdysteroid titers were positively correlated, and were lowest when a queen was present. In queenless workers, ecdysteroid levels were elevated in dominant workers, and were also influenced by the presence of brood and by group demography; hormone levels were higher in bees kept in larger groups. These findings are consistent with the premise that in B. terrestris the ovary is the primary site of ecdysteroid synthesis, and they show that ecdysteroids levels vary with the social environment. PMID- 15885701 TI - Cold hardiness of Habrobracon hebetor (Say) (Hymenoptera: Braconidae), a parasitoid of pyralid moths. AB - The ectoparasitoid Habrobracon hebetor (Say) attacks stored-product infesting pyralid moths that are able to overwinter under extremely cold conditions. The extent to which H. hebetor can withstand these conditions is not known, but has important implications for the ability of H. hebetor to provide long-term suppression of these pests in temperate climates. We investigated basic cold hardiness aspects of a mutant eye-color strain of H. hebetor. Feeding larvae and adults of H. hebetor had supercooling points (SCPs) at temperatures higher than those of eggs and pupae. Mean SCPs of females and males were equivalent, as were those of naked and silk-encased pupae. Feeding on honey prior to being subjected to low temperatures significantly increased the SCP of adult females by approximately 8 degrees C. Mortality of pupae and adults increased significantly whenever the temperature dropped below the mean SCP, indicating that H. hebetor does not tolerate freezing. For pupae and adults exposed to -12 and -5 degrees C, the hourly mortality rate increased with time of exposure. Pupae and adults exposed to -12 degrees C for different time intervals showed high mortality after only 1d of exposure. At -5 degrees C, none survived 12d of exposure. A better understanding of how well this parasitoid tolerates low temperatures will be useful in evaluating its potential as a biological control agent of stored product moths in temperate regions. PMID- 15885702 TI - Diversity in hapten recognition: structural study of an anti-cocaine antibody M82G2. AB - Antibodies against cocaine and other drugs of abuse are the basis for diagnostic tests for the presence of those drugs in human serum. The 1.7A resolution crystal structure of the anti-cocaine monoclonal antibody M82G2 in complex with cocaine is presented. This structure determination was undertaken to establish the stereochemical features in the antibody binding site that confer specificity for cocaine, and as part of an ongoing project to understand the rules that govern molecular recognition. The cocaine-binding site can be characterized topologically as a narrow groove on the protein surface. The antibody utilizes water-mediated hydrogen bonding, and cation-pi and stacking (pi-pi) interactions to provide specificity. Comparison with the previously published structure of the anti-cocaine antibody GNC92H2 shows that binding of a small ligand can be achieved in diverse ways, both in terms of a binding site structure/topology and protein-ligand interactions. PMID- 15885704 TI - Non-fatal transportation injuries among women: differences in injury patterns and severity by age. PMID- 15885705 TI - The crash severity impacts of fixed roadside objects. AB - INTRODUCTION: This study analyzes the in-service performance of roadside hardware on the entire urban State Route system in Washington State by developing multivariate statistical models of injury severity in fixed-object crashes using discrete outcome theory. The objective is to provide deeper insight into significant factors that affect crash severities involving fixed roadside objects, through improved statistical efficiency along with disaggregate and multivariate analysis. METHOD: The developed models are multivariate nested logit models of injury severity and they are estimated with statistical efficiency using the method of full information maximum likelihood. RESULTS: The results show that leading ends of guardrails and bridge rails, along with large wooden poles (e.g. trees and utility poles) increase the probability of fatal injury. The face of guardrails is associated with a reduction in the probability of evident injury, and concrete barriers are shown to be associated with a higher probability of lower severities. Other variables included driver characteristics, which showed expected results, validating the model. For example, driving over the speed limit and driving under the influence of alcohol increase the probability of fatal accidents. Drivers that do not use seatbelts are associated with an increase in the probability of more severe injuries, even when an airbag is activated. IMPACT ON INDUSTRY: The presented models show the contribution of guardrail leading ends toward fatal injuries. It is therefore important to use well-designed leading ends and to upgrade badly performing leading ends on guardrails and bridges. The models also indicate the importance of protecting vehicles from crashes with rigid poles and tree stumps, as these are linked with greater severities and fatalities. PMID- 15885706 TI - An analytical study of the number of steady states in gene regulatory networks. AB - Herein, I proposed a model for gene networks and studied the steady states in the dynamics both by numerical and analytical methods. In this model, mRNA and protein levels change continuously with time; each gene alters transcriptional regulation depending on the concentration of transcription factors. The dynamical behavior of continuous model is quite complex and different from that of the discrete model, the Boolean network. Large portion of steady states of this model can be classified into three types. The rough structure of gene interactions, which corresponds to Boolean function, is sufficient to predict the expression level of each gene in these types of steady states. I also determined the expected numbers of two major types of steady states observed in a randomly generated gene network. The results obtained from these formulae contradict previously accepted belief. The results are that neither gene number nor connectivity between genes increases the expected number of steady states in a random gene network. The number of steady states is very small. The number of self-regulatory genes, however, effectively increases the number of steady states in a network. These results imply that increases in gene number may not be the direct driving force for the evolution of a variety of different cell types within organisms. Instead, the number of self-regulatory genes may significantly increase cellular diversity. PMID- 15885707 TI - Chemomodulatory effect of Ficus racemosa extract against chemically induced renal carcinogenesis and oxidative damage response in Wistar rats. AB - Ferric nitrilotriacetate (Fe-NTA) is a well-known renal carcinogen. In this communication, we show the chemopreventive effect of Ficus racemosa extract against Fe-NTA-induced renal oxidative stress, hyperproliferative response and renal carcinogenesis in rats. Fe-NTA (9 mg Fe/kg body weight, intraperitoneally) enhances renal lipid peroxidation, xanthine oxidase, gamma-glutamyl transpeptidase and hydrogen peroxide (H(2)O(2)) generation with reduction in renal glutathione content, antioxidant enzymes, viz., glutathione peroxidase, glutathione reductase, catalase, glucose-6-phosphate dehydrogenase and phase-II metabolising enzymes such as glutathione-S-transferase and quinone reductase. It also enhances blood urea nitrogen, serum creatinine, ornithine decarboxylase (ODC) activity and thymidine [(3)H] incorporation into renal DNA. It also enhances DEN (N-diethylnitrosamine) initiated renal carcinogenesis by increasing the percentage incidence of tumors. Treatment of rats orally with F. racemosa extract (200 and 400 mg/kg body weight) resulted in significant decrease in gamma glutamyl transpeptidase, lipid peroxidation, xanthine oxidase, H(2)O(2) generation, blood urea nitrogen, serum creatinine, renal ODC activity, DNA synthesis (P<0.001) and incidence of tumors. Renal glutathione content (P<0.01), glutathione metabolizing enzymes (P<0.001) and antioxidant enzymes were also recovered to significant level (P<0.001). Thus, our data suggests that F. racemosa extract is a potent chemopreventive agent and suppresses Fe-NTA-induced renal carcinogenesis and oxidative damage response in Wistar rats. PMID- 15885708 TI - Overweight is gender-dependent in prenatal protein--calorie restricted adult rats acting on the blood pressure and the adverse cardiac remodeling. AB - Postnatal heart remodeling was studied in rats submitted to prenatal protein- calorie restriction (R). Offspring were divided in four groups: control male (CM) and female (CF) vs. restricted male (RM) and female (RF) and lived 120 days. The offspring blood pressure (BP) and biometry were periodically analyzed. In the euthanasia day, the left ventricular (LV) mass index, the cardiomyocyte nuclei profile number (N[cmn]) (disector method), the cross-sectional cardiomyocyte area (A[cm]) and the stereology for intramyocardial arteries (ima) were estimated. Interactions between gender and prenatal nutritional conditions were tested with the two-way ANOVA. RM animals showed higher BP and greater body mass and smaller LV mass index than the other groups. N[cmn] and stereology parameters of ima were smaller, and A[cm] was greater in the R groups rats than in the C groups rats; these structural changes were only dependent of the prenatal nutritional condition but not gender-dependent. IN CONCLUSION: hypertension and body and cardiac growth were influenced by the interaction between gender and prenatal nutritional conditions, while cardiac remodeling seems to be only programmed by the adverse intrauterine environment. PMID- 15885709 TI - Decay model for biocide treatment of unballasted vessels: application for the Laurentian Great Lakes. AB - A biocide decay model was developed to assess the potential efficacy and environmental impacts associated with using glutaraldehyde to treat unballasted overseas vessels trading on the Laurentian Great Lakes. The results of Monte Carlo simulations indicate that effective glutaraldehyde concentrations can be maintained for the duration of a vessel's oceanic transit (approximately 9-12 days): During this transit, glutaraldehyde concentrations were predicted to decrease by approximately 10% from initial treatment levels (e.g., 500 mgL(-1)). In terms of environmental impacts, mean glutaraldehyde concentrations released at Duluth-Superior Harbor, MN were predicted to be 100-fold lower than initial treatment concentrations, and ranged from 3.2 mgL(-1) (2 SD: 2.74) in April to 0.7 mgL(-1) (2 SD: 1.28) in August. Sensitivity analyses indicated that the re ballasting dilution factor was the major variable governing final glutaraldehyde concentrations; however, lake surface temperatures became increasingly important during the warmer summer months. PMID- 15885710 TI - In pursuit of effective toxicogenomics. AB - Biological systems exhibit complex responses to xenobiotics varying from generic stress responses to very specific changes closely associated with the mechanism of toxicity. Until recently our view of this complexity was obscured by the simplicity of available analysis tools which allowed determination of only a few genes in any one study. Then genome sequencing and high throughput library screening projects delivered data on the genome sequence of many organisms, and clones were collected and made available to researchers in a previously unparalleled quantity. To exploit this new resource the microarray was developed from its predecessor the dot blot. Further development has expanded the number of clones contained on any one microarray to a point where the expression of many tens of thousands of genes in a biological system can be determined in a short period of time. What these data are revealing is the full complexity of the gene expression response to stimuli such as xenobiotic exposure. Toxicogenomics seeks to use the complexity of this response as a fingerprint or signature characteristic of that xenobiotic exposure. There are though two major experimental challenges that need to be dealt with for toxicogenomics to be successful. The first is technical and relates to the intrinsic difficulties associated with the accurate measurement of gene expression. For microarrays, this problem is multiplied by the number of genes on the microarray itself. To overcome this technical variability correct experimental design is critical. The second challenge concerns the biological system used. What genetic background, time point and dose of xenobiotic should be chosen? For in vitro systems should cell lines or primary cells be used? These factors, and more, could affect the gene expression profile obtained in response to the same xenobiotic exposure. Using both our data and data from public databases these issues are explored in this paper. PMID- 15885711 TI - Okadaic acid: chromosomal non-disjunction analysis in human lymphocytes and study of aneugenic pathway in CHO-K1 cells. AB - Okadaic acid (OA) is the main marine toxin implicated in the diarrhetic shellfish poisoning (DSP) in humans after consumption of contaminated bivalve molluscs. We have previously shown that OA was an in vitro aneugenic compound that induced chromosome loss via micronuclei formation in CHO-K1 cells. The aims of this study were to investigate the chromosomal non-disjunction (ND) potential of OA in human lymphocytes and the pathways involved for aneuploidy in CHO-K1 cells. Firstly, we analysed the formation of micronuclei and the non-disjunction for chromosomes 1 and 17 in binucleated human lymphocytes cells with the cytokinesis-blocked micronucleus (CBMN) assay coupled to a fluorescent in situ hybridization (FISH) technique with centromere-specific DNA probes. We showed that OA statistically increased the frequency of micronucleated lymphocytes in the dose range from 20 to 35 nM. However, FISH analysis did not reveal any increase in the non disjunction for both chromosomes whatever the concentration between 2.5 and 35 nM. However, a significant increase in ND for the chromosome 17 was found at 1 nM. Secondly, in CHO-K1 cells, we investigated the dose and time dependent effects of OA: (i) on cell cycle progression, (ii) on mitotic-phase arrest and (ii) on mitotic spindle and centrosome abnormalities. The results showed that OA induced a progressive accumulation of mitotic CHO-K1 cells in prometaphase, an induction of multipolar mitotic spindle with centrosome amplification and the formation of multinucleated cells. We concluded that OA did not induce chromosome non-disjunction but should more likely induced chromosome loss in human lymphocytes. Moreover, our results obtained in CHO-K1 suggest that OA induced aneuploidy by preventing the chromosome attachment to the mitotic spindle and by amplifying the centrosome. The mode of action of the toxin in relation to its inhibition of protein phosphatases 1 (PP1) and 2A (PP2A) and the mitosis process is discussed. PMID- 15885712 TI - HIM1, a new yeast Saccharomyces cerevisiae gene playing a role in control of spontaneous and induced mutagenesis. AB - We have identified a new Saccharomyces cerevisiae gene, HIM1, mapped on the right arm of the chromosome IV (ORF YDR317w), mutations in which led to an increase in spontaneous mutation rate and elevated the frequencies of mutations, induced by UV-light, nitrous acid, ethylmethane sulfonate and methylmethane sulfonate. At the same time, him1 mutation did not result in the increase of the sensitivity to the lethal action of these DNA-damaging agents. We tested the induced mutagenesis in double mutants carrying him1 mutation and mutations in other repair genes: apn1, blocking base excision repair; rad2, rev3, and rad54, blocking three principal DNA repair pathways; pms1, blocking mismatch repair; hsm2 and hsm3 mutations, which lead to a mutator effect. Epistatic analysis showed a synergistic interaction of him1 with pms1, apn1, and rad2 mutations, and epistasis with the rev3, the rad54, the hsm2, and the hsm3. To elucidate the role of the HIM1 in control of spontaneous mutagenesis, we checked the repair of DNA mispaired bases in the him1 mutant and discovered that it was not altered in comparison to the wild-type strain. In our opinion, our results suggest that HIM1 gene participates in the control of processing of mutational intermediates appearing during error-prone bypass of DNA damage. PMID- 15885713 TI - The use of genetically engineered mouse models of prostate cancer for nutrition and cancer chemoprevention research. AB - The ability to modify the expression of specific genes in the mouse through genetic engineering technologies allows for the generation of previously unavailable models for prostate cancer prevention research. Although animal models have existed for some time for the study of prostate cancer prevention (primarily in the rat), it is uncertain if the mechanisms that drive prostate carcinogenesis in these models are relevant to those in human prostate cancer. Cell culture studies are of limited usefulness because the conditions are inherently artificial. Factors such as relevant physiologic concentrations and metabolism of putative chemoprevention compounds are difficult to model in an in vitro system. These studies also preclude the types of interactions known to occur between multiple cell types in vivo. In addition, all prostate cancer cell lines are already highly progressed and are not representative of the type of cells to which most preventive strategies would be targeted. Due to the advent of genetically engineered mouse (GEM) models, we now have models of prostate cancer that are dependent on molecular mechanisms already implicated in human prostate carcinogenesis. With these models we can perform a variety of experiments that could previously only be done in cell culture or in prostate cancer cell line xenografts. The currently available GEM models of prostate cancer have been extensively reviewed therefore, this review will focus on the types of models available and their usefulness for various types of preclinical studies relevant to prostate cancer prevention. PMID- 15885714 TI - Therapeutic opportunities through modulation of the endocannabinoid system. PMID- 15885715 TI - Foreword by Igor Grant, M.D., Director, Center for Medicinal Cannabis Research (CMCR). PMID- 15885716 TI - Right parietal contributions to verbal working memory: spatial or executive? AB - The left inferior parietal cortex has been claimed to be the site of the verbal short-term store, yet imaging studies report activity of a homologous right hemisphere region in verbal working memory tasks as well. In spite of its prevalent activity, right parietal contributions to verbal working memory are poorly understood. To clarify its role in verbal working memory performance, we tested a patient with a lesion in the right parietal lobe on verbal and spatial versions of the N-back task. The patient was impaired in all the spatial conditions regardless of load (0-, 1-, and 2-back), whereas in the verbal N-back he was impaired only in the conditions with a memory demand (1- and 2-back). Given that we had presented stimuli at multiple locations in the verbal N-back, however, it remained possible that the lesion impaired spatial representation rather than verbal working memory per se. With central stimulus presentation, his performance dramatically improved indicating that his difficulty with the N-back task was largely due to his poor visuospatial abilities. PMID- 15885717 TI - What is the locus of the errorless-learning advantage? AB - In two experiments involving word-stem completion, an advantage was found for errorless over errorful-learning conditions, for both severely and moderately memory-impaired participants. This advantage did not depend on the implicit/explicit nature of the question asked. Additional tests showed that subsequent recognition of target items was good for both groups, but only in the absence of lures derived from participants' prior errors. Source-memory was shown to be virtually absent in the severely impaired group and only weakly present in the moderately impaired group. This combination of results suggests that preserved implicit memory, in the absence of explicit memory, is sufficient for an errorless-learning advantage to accrue. PMID- 15885718 TI - Conditional discrimination and reversal in amnesia subsequent to hypoxic brain injury or anterior communicating artery aneurysm rupture. AB - Human anterograde amnesia can develop following bilateral damage to the hippocampus and medial temporal lobes, as in hypoxic brain injury, or following damage to the basal forebrain, as following anterior communicating artery (ACoA) aneurysm rupture. In both cases, the mnestic deficit may be similar when assessed by standard neuropsychological measures. However, animal and computational models suggest that there are qualitative differences in the pattern of impaired and spared memory abilities following damage to hippocampus versus basal forebrain. Here, we show such a dissociation in human amnesia using a single two-stage task, involving conditional discrimination and reversal. Consistent with a prior study, 10 individuals with anterograde amnesia subsequent to hypoxic brain injury were spared on acquisition but impaired at reversal. However, 10 individuals with amnesia subsequent to ACoA aneurysm showed the opposite pattern of impaired acquisition but spared reversal. The differences between groups cannot be easily ascribed to severity of mnestic or cognitive deficit, since the two amnesic groups performed similarly on neuropsychological tests of memory, intelligence and attention. The results illustrate qualitative differences in memory impairments in hypoxic and ACoA amnesics and highlight the importance of considering etiology in evaluating mnemonic deficits in amnesic populations. PMID- 15885719 TI - Serotonin type-1A receptors modulate adolescent, cocaine-induced offensive aggression in hamsters. AB - Hamsters repeatedly exposed to cocaine throughout adolescence display highly escalated offensive aggression compared to saline-treated littermates. Recently, we have shown that serotonin neural signaling and development play an important role in adolescent cocaine-induced offensive aggression. This study examined whether the adolescent cocaine-induced aggressive response was modulated by serotonin type 1A (5HT1A) receptors. To test this, adolescent male Syrian hamsters were administered cocaine hydrochloride (0.5 mg/kg, i.p.) throughout adolescent development (P27-57) and then tested for offensive aggression after the administration of the 5HT(1A) receptor agonist R(+)-8-OH-DPAT (0.1, 0.3, 0.6, 1.0, 1.25 mg/kg, i.p.). R(+)-8-OH-DPAT dose-dependently reduced cocaine-induced offensive aggression, with a significant reduction observed at 0.3 mg/kg for most of the offensive responses measured. Animals treated with higher doses of R(+)-8 OH-DPAT (0.6-1.25 mg/kg) prior to testing showed significant reductions in all measures of offensive aggression and social interest towards intruders (i.e., contact time), indicating more general behavioral inhibition. Adolescent cocaine treated animals did not differ in body weight from controls, suggesting that the increased aggression was not due to increased body mass. These data support a role for 5HT1A signaling in adolescent cocaine-induced aggression. PMID- 15885720 TI - The cost of a specific immune response in young guinea pigs. AB - The specific immune system is a protective mechanism that detects infection and fights it by production of antibodies. Newborns are especially susceptible to infections because their immune system is not yet as fully developed as that of adults. This has been well established in altricial mammals. Fighting infection is associated with costs (metabolic rate, protein synthesis) potentially affecting other developmental processes. We investigated the specific immune response in a precocial mammal, by testing the response of 3 and 7 day old young guinea pigs (Cavia aperea f. porcellus) against a non-pathogenic antigen (KLH) and determined the effect of the immune response on growth and metabolic rate. Challenged young produced a substantial specific immune response (IgG). The efficiency of the immune response was almost identical in 3 and 7 day old young, but lower than in adult females. Antibody titres achieved by actively immunised young pups were as high as titres transferred transplacentally by mothers immunised on day 40 and 47 of pregnancy. In comparison to a control group, the immune response did not influence growth and metabolic rate measured on day 4 after each immune challenge and was not reflected by changes in hematocrit value. We discuss whether the weaker immune response of pups is caused by reduced allocation of limited resources in growing young or by the immature immune system of young animals. PMID- 15885721 TI - Measuring socio-economic inequalities in the presentation of injuries to a paediatric A&E department: the importance of an epidemiological approach. AB - OBJECTIVES: To contrast the socio-economic pattern of childhood injuries presenting to a paediatric accident and emergency (A&E) department revealed by using both a numerator-based and a denominator-based approach to the analysis of injury surveillance data. METHODS: Injury surveillance data collected during 1997 1998 at a Glasgow children's hospital A&E department were analysed. Socio economic status was measured using Carstairs' deprivation index. Data from West Glasgow postcode sectors only were analysed in order to optimize epidemiological validity. Socio-economic patterning of injury was investigated in two ways numerator-based and denominator-based. RESULTS: A total of 12,762 children (0-14 years) living in West Glasgow attended the A&E department of the Royal Hospital for Sick Children over the study period. Both analytical approaches showed a clear and statistically significant excess of injury presentations in children from more deprived postcode sectors, but the variation appeared much greater in the numerator-based rather than the denominator-based approach. In regression analysis, however, only the denominator-derived rates showed a statistically significant linear trend across deprivation categories. CONCLUSION: The most appropriate and accurate means of measuring the extent of socio-economic (and other) inequalities in injury risk is to adopt a population-based rather than numerator-based perspective on the data collected by injury surveillance systems. PMID- 15885722 TI - Population mobility: characteristics of people registering with general practices. AB - OBJECTIVES: The aim of this study was to examine the characteristics of patients joining general practitioners' (GP) lists, and the time taken to register after a move of residence. STUDY DESIGN: Questionnaire study. METHODS: Staff in six London general practices administered the questionnaire to 642 newly registering adults. RESULTS: Nearly 40% of participants took longer than 6 months to re register with a GP after a change of address. About one in eight participants (13%) took longer than 1 year and one in 14 (7%) took longer than 3 years to register. The overall median time to register after a move was 4 months. The amount of time taken to register appeared to be influenced by a number of factors, including gender, age and geographical location. CONCLUSIONS: Population mobility and the time taken to register with a new GP is likely to have a major impact on access to health care and the effectiveness of local preventative health programmes. Primary care trusts need to encourage their local residents to register with a GP soon after a change of address, and develop initiatives to encourage participation in preventative health programmes amongst mobile groups. Additional measures to strengthen primary care provision, such as walk-in centres, may be required in areas with the highest levels of population turnover. PMID- 15885723 TI - Health care costs and financial consequences of epidemiological changes in chronic diseases in Latin America: evidence from Mexico. AB - OBJECTIVE: To determine the costs of health services and the financial consequences of changes in the epidemiological profile of chronic diseases in Latin America. STUDY DESIGN: We conducted longitudinal analyses of costs and of the economic impact of the epidemiological transition in healthcare services for diabetes and hypertension in the Mexican health system. The study population included both the insured and uninsured populations. METHODS: The cost-evaluation method was based on the instrumentation and consensus techniques. To estimate the epidemiological changes and financial consequences for 2004-2006, six models were constructed according to the Box-Jenkins technique, using confidence intervals of 95% and the Box-Pierce test. FINDINGS: Costs ranged from US$613 to US$887 for diabetes, and from US$485 to US$622 for hypertension. Regarding epidemiological changes for 2004 compared with 2006, an increase is expected in both cases, although results predict a greater increase for diabetes, 10-15% in all three institutions (P<0.05). Comparing the financial consequences of health services required by insured and uninsured populations, the greater increase (17%) will be for the insured population (P<0.05). The financial requirements for both diseases will amount to 9.5% of the total budget for the uninsured population and 13.5% for the insured population. CONCLUSIONS: If the risk factors and the different healthcare models remain as they are at present, the economic impact of expected epidemiological changes on the social security system will be particularly strong. Another relevant financial factor is the appearance of internal competition in the use and allocation of financial resources among the main providers in the health services; this factor becomes even more complicated within each provider. In effect, within each institution, hypertension and diabetes programmes must compete for resources with other programmes for chronic and infectious diseases. PMID- 15885724 TI - Alcohol prevention measures at an emergency department: physicians' perspectives. PMID- 15885725 TI - Naturally acquired Lawsonia intracellularis infection in pigs studied from weaning to slaughter by indirect immunofluorescence antibody test and polymerase chain reaction on faeces. AB - The course of naturally acquired Lawsonia intracellularis infection was studied in 41 pigs by testing blood and faeces samples collected four to seven times from before weaning to slaughter 5 months old. At slaughter, a sample of ileum was taken for histopathology. In the first sampling when the pigs were 2-4 weeks old maternally derived IgG against L. intracellularis was demonstrated by immunofluorescence antibody test in nine pigs whereas the bacterium was detected by PCR in faeces from six pigs. The maternally derived antibodies did not prevent pigs from becoming infected as seven pigs later on shed and/or were seropositive for L. intracellularis. The lowest prevalence of L. intracellularis was observed in 6-13 weeks old pigs and it seemed as though L. intracellularis in early infected pigs only activates a minor antibody response. At slaughter 66% of the pigs were found positive by immunofluorescence antibody test compared to 24% by immunohistochemistry on ileal samples. Thus, applied at the time of slaughter the antibody test appeared to be a highly sensitive ante-mortem diagnostic tool for identifying L. intracellularis exposed pigs with or without current proliferative enteropathy. PMID- 15885726 TI - C-25 epimeric 26-haloponasterone A: synthesis, absolute configuration and moulting activity. AB - A convenient synthesis of inokosterone has been accomplished. Inokosterone exists as two C-25 epimers, which could be separated from each other through their diacetonide derivatives. The absolute configuration of these compounds was determined. Two C-25 epimers of 26-chloroponasterone A were synthesized from the respective C-25 epimeric inokosterone. Two epimeric 26-bromo and 26 iodoponasterone A compounds were also synthesized. Moulting activity of these compounds was evaluated using the Musca bioassay, and it was found that the (25S) 26-halo analogues were more active than the corresponding (25R)-26-halo analogues. Among the 25S series, an increase in activity with an increase in size of the halogen atom was observed, indicating that the steric factor was more important than the electronic factor in binding of these ecdysteroid analogues to the receptor. On the other hand, a decrease in activity with an increase in size of the halogen atom was noted in the 25R series, suggesting that the steric factor was less important than the electronic factor. The results indicated that the configuration at C-25 and the substituent at C-26 have significant influences on the interaction of ecdysteroids with their receptor. PMID- 15885727 TI - Effects of oral estrone on rat energy balance. AB - Oral doses of estrone from 10 nmol/(kg day) to 10 micromol/(kg day) were given to adult Wistar male rats for 10 days. Body composition, energy balance, total body estrone balance and plasma metabolites and hormones were measured at the end of the treatment. Body weight (as well as food intake, body energy, fat and water accrual) increased at doses in the 10--100 nmol/(kg day) range, but decreased at higher doses. Energy expenditure decreased with increasing doses of estrone. Plasma metabolite changes suggested the maintenance of energy homeostasis, and lipid parameters indicated that lipid mobilization increased with the increasing doses of estrone. Plasma estrone, acyl-estrone and estradiol levels decreased at low doses and increased at high doses of estrone. We conclude that: (a) repeated estrone gavages, even at very high doses, do not result in the accrual of estrone in the body; (b) low doses of estrone promote growth and high doses decrease body mass and fat accretion; (c) administration of estrone at low doses decreases its circulating levels and the levels of estradiol and acyl-estrone, a situation reverted at higher doses and (d) estrone administration induces a dose-dependent shift towards lower energy expenditure. PMID- 15885728 TI - Morphologic changes in alveolar macrophages in response to UVEC-activated pulmonary Type II epithelial cells. AB - We hypothesize that Type II epithelial cells, which line the distal airspaces of the lung, are early responders to invading pathogens and release a signal, which activates and alters the phenotype and phagocytosis properties of alveolar macrophages even at a distance. The T(7) cell line is a conditionally immortalized murine Type II epithelial cell line developed in our laboratory. Using an in vitro transwell model we have previously shown that UV-irradiated Escherichia coli (UVEC)-stimulated T(7) cells cultured in the lower transwell chamber, release a diffusible signal which activates MH-S cells (immortalized murine alveolar macrophages) cultured in the upper transwell chamber, to produce nitric oxide. Using scanning electron microscopy, we show that MH-S cells activated in this manner exhibit increased cell surface ruffling, numerous long filopodia, increased lamellipodia and cell flattening. DynaBead uptake studies show that these morphologic changes are accompanied by increased phagocytosis. These findings indicate that a diffusible signal released at a distance by UVEC stimulated Type II epithelial cells initiates changes in morphology and phagocytosis reflective of macrophage activation concomitant with the functional activation we previously reported. PMID- 15885729 TI - Lignans, bacteriocides and organochlorine compounds activate the human pregnane X receptor (PXR). AB - The pregnane X receptor (PXR) mediates the induction of enzymes involved in steroid metabolism and xenobiotic detoxification. The receptor is expressed in liver and intestinal tissues and is activated by a wide range of compounds. The ability of a diverse range of dietary compounds to activate PXR-mediated transcription was assayed in HuH7 cells following transient transfection with human PXR (hPXR). The compounds investigated included phytochemicals such as lignans and phytoestrogens, organochlorine dietary contaminants such as polychlorinated biphenyls (PCBs) and triclosan and selected steroid, drug and herbal compounds. The hPXR activation at the top concentrations tested (10 microM) relative to the positive control 10 microM rifampicin ranged from 1.3% (trans-resveratrol) to 152% (ICI 182780). Hydroxylated compounds were marginally more potent than the parent compounds (tamoxifen activation was 74.6% whereas 4 hydroxytamoxifen activation was 84.2%) or significantly greater (vitamin D3 activation was 1.6%, while hydroxylated vitamin D3 activation was 55.6%). Enterolactone, the metabolite of common dietary lignans, was a medium activator of PXR (35.6%), compared to the lower activation of a parent lignan, secoisolariciresinol (20%). Two non-hydroxylated PCB congeners (PCB 118 and 153), which present a larger fraction of the PCB contamination of fatty foods, activated hPXR by 26.6% and 17%, respectively. The pesticide trans-nonachlor activation was 53.8%, while the widely used bacteriocide triclosan was a medium activator of hPXR at 46.2%. The responsiveness of PXR to activation by lignan metabolites suggests that dietary intake of these compounds may affect the metabolism of drugs that are CYP3A substrates. Additionally, the evidence that organochlorine chemicals, particularly the ubiquitous triclosan, activate hPXR suggests that these environmental chemicals may, in part, exhibit their endocrine disruptor activities by altering PXR-regulated steroid hormone metabolism with potential adverse health effects in exposed individuals. PMID- 15885730 TI - Role of osteopontin in hepatic neutrophil infiltration during alcoholic steatohepatitis. AB - Alcoholic liver disease (ALD) is a major complication of heavy alcohol (EtOH) drinking and is characterized by three progressive stages of pathology: steatosis, steatohepatitis, and fibrosis/cirrhosis. Alcoholic steatosis (AS) is the initial stage of ALD and consists of fat accumulation in the liver accompanied by minimal liver injury. AS is known to render the hepatocytes increasingly sensitive to toxicants such as bacterial endotoxin (LPS). Alcoholic steatohepatitis (ASH), the second and rate-limiting step in the progression of ALD, is characterized by hepatic fat accumulation, neutrophil infiltration, and neutrophil-mediated parenchymal injury. However, the pathogenesis of ASH is poorly defined. It has been theorized that the pathogenesis of ASH involves interaction of increased circulating levels of LPS with hepatocytes being rendered highly sensitive to LPS due to heavy EtOH consumption. We hypothesize that osteopontin (OPN), a matricellular protein (MCP), plays an important role in the hepatic neutrophil recruitment due to its enhanced expression during the early phase of ALD (AS and ASH). To study the role of OPN in the pathogenesis of ASH, we induced AS in male Sprague-Dawley rats by feeding EtOH-containing Lieber DeCarli liquid diet for 6 weeks. AS rats experienced extensive fat accumulation and minimal liver injury. Moderate induction in OPN was observed in AS group. ASH was induced by feeding male Sprague-Dawley rats EtOH-containing Lieber-DeCarli liquid diet for 6 weeks followed by LPS injection. The ASH rats had substantial neutrophil infiltration, coagulative oncotic necrosis, and developed higher liver injury. Significant increases in the hepatic and circulating levels of OPN was observed in the ASH rats. Higher levels of the active, thrombin-cleaved form of OPN in the liver in ASH group correlated remarkably with hepatic neutrophil infiltration. Finally, correlative studies between OPN and hepatic neutrophil infiltration was corroborated in a simple rat peritoneal model where enhanced peritoneal fluid neutrophil infiltration was noted in rats injected OPN intraperitoneally. Taken together these data indicate that OPN expression induced during ASH may play a significant role in the pathogenesis of ASH by stimulating neutrophil transmigration. PMID- 15885731 TI - Lead enhances CD4+ T cell proliferation indirectly by targeting antigen presenting cells and modulating antigen-specific interactions. AB - Although Pb is a well-known immunotoxicant, its mechanism of action is not well understood. Low levels of Pb (approximately 1 microM) markedly enhance the proliferative T cell response in mixed lymphocyte culture (MLC), a process we have termed allo-enhancement. As Pb allo-enhancement occurs whether alloantigen presenting cells (APC) are derived from C57BL/6 or BALB.B10, the allo-reactive T cells involved are likely to be specific for peptide in the context of the IA(b) molecule as the IE molecule is null in H-2(b) mice. Analysis of T cell division in MLC with Pb treatment indicated that there was no significant difference between Pb and non-Pb-treated cultures until day 4 when the frequency of proliferating T cells was much greater than in non-treated cultures. Our data suggest that this increased proliferation is not coupled with increased IL-2 levels in the media as these were actually decreased with Pb treatment and that Pb-induced enhancement in the allo-proliferative response is only partially dependent upon IL-2. Pb allo-enhancement is abrogated when stimulating allo-APCs are paraformaldehyde-fixed, and T cell proliferation stimulated by concanavalin A is not enhanced with Pb treatment, suggesting that the APC is the proximate target of Pb in allo-MLC. Pb allo-enhancement does not occur when T cells respond to irradiated allo-B cells, alone; however, it is restored when syngeneic CD11c enriched cells are added. Of the CD11c-enriched splenocytes, the fraction that is adherent after 24 h, consistent with macrophages, appears to be the cell type targeted by Pb. Using T cells from DO11.10 transgenic mice, we determined that the effect of Pb is centered around specific p:MHC interactions and that enhanced costimulation is an unlikely mechanism for Pb allo-enhancement. PMID- 15885732 TI - Low dose DDT inhibition of hepatocarcinogenesis initiated by diethylnitrosamine in male rats: possible mechanisms. AB - Previously we reported a tendency for reduction of the development of glutathione S-transferase placental form (GST-P) positive foci, recognized as preneoplastic changes in rat liver, by a low dose of 1,1-bis(p-chlorophenyl)-2,2,2 trichloroethane (DDT), which belongs to the same group of hepatic cytochrome P 450 inducers as phenobarbital and is itself a non-genotoxic hepatocarcinogen. In order to clarify the biological significance of this phenomenon, we investigated the reproducibility and changes in other parameters using an initiation-promotion model in which male F344 rats were treated with DDT at doses of 0, 0.005, 0.5, 500 ppm in the diet for 11 or 43 weeks after initiation of hepatocarcinogenesis with N-diethylnitrosamine (DEN). When 500 ppm DDT was applied, the formation of GST-P positive foci and tumor were markedly elevated. In contrast, induction of GST-P positive foci and liver tumors tended to be inhibited at a dose of 0.005 ppm, correlating with protein levels of cytochrome P450 2B1 and 3A2 (CYP2B1 and 3A2) and generation of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a marker of oxidative DNA damage. mRNA levels for 8-oxoguanine glycosylase 1 (OGG1), an 8 OHdG repair enzyme, connexin 32 (Cx32), a major component of Gap junctions, and hepatic nuclear factor 1alpha (HNF-1alpha), a Cx32 regulator, were inversely correlated with GST-P positive foci and tumor formation. These results indicate that low dose DDT may indeed exhibit inhibitory effects on chemically initiated rat hepatocarcinogenicity, in contrast to the promotion observed with high doses, and that this is related to changes in metabolizing enzymes, cell communication, and DNA damage and its repair. PMID- 15885733 TI - Cytogenetic damage in lymphocytes of patients undergoing therapy for small cell lung cancer and ovarian carcinoma. AB - The level of cytogenetic damage in peripheral blood lymphocytes of patients undergoing chemotherapy has been analyzed incisively 20 years ago. The results showed that the highest level of cytogenetic damage was observed at the end of therapy. In recent years, the doses of anticancer drugs were intensified thanks to the discovery of colony stimulating factors. Therefore, it was interesting to analyze the kinetics of micronuclei formation in lymphocytes of patients undergoing modern chemotherapy. The frequencies of micronuclei were measured in lymphocytes of 6 patients with small cell lung cancer treated with a combination of cisplatin and etoposide and 7 patients with ovarian carcinoma treated with a combination of taxol and cisplatin. 3 patients with lung cancer received radiotherapy in addition to chemotherapy. Micronuclei were analyzed in lymphocytes collected before the start of therapy and 1 day before each following cycle of chemotherapy. The micronucleus frequencies were compared with the kinetics of leukocyte counts. The micronucleus frequencies showed an interindividual variability. On average, the frequencies of micronuclei increased during the first half of therapy and declined thereafter, reaching, in some patients with ovarian carcinoma, values below the pre-treatment level. Leukocyte counts decreased strongly at the beginning of therapy with an upward trend at the end. We suggest that the decline of micronuclei was due to repopulation of lymphocytes and acquired drug resistance. PMID- 15885734 TI - Induction of Cyp1a1 and Cyp1b1 and formation of DNA adducts in C57BL/6, Balb/c, and F1 mice following in utero exposure to 3-methylcholanthrene. AB - Fetal mice are more sensitive to chemical carcinogens than are adults. Previous studies from our laboratory demonstrated differences in the mutational spectrum induced in the Ki-ras gene from lung tumors isolated from [D2 x B6D2F1]F2 mice and Balb/c mice treated in utero with 3-methylcholanthrene (MC). We thus determined if differences in metabolism, adduct formation, or adduct repair influence strain-specific responses to transplacental MC exposure in C57BL/6 (B6), Balb/c (BC), and reciprocal F1 crosses between these two strains of mice. The induction of Cyp1a1 and Cyp1b1 in fetal lung and liver tissue was determined by quantitative fluorescent real-time PCR. MC treatment caused maximal induction of Cyp1a1 and Cyp1b1 RNA 2-8 h after injection in both organs. RNA levels for both genes then declined in both fetal organs, but a small biphasic, secondary increase in Cyp1a1 was observed specifically in the fetal lung 24-48 h after MC exposure in all four strains. Cyp1a1 induction by MC at 4 h was 2-5 times greater in fetal liver (7000- to 16,000-fold) than fetal lung (2000- to 6000-fold). Cyp1b1 induction in both fetal lung and liver was similar and much lower than that observed for Cyp1a1, with induction ratios of 8- to 18-fold in fetal lung and 10- to 20-fold in fetal liver. The overall kinetics and patterns of induction were thus very similar across the four strains of mice. The only significant strain-specific effect appeared to be the relatively poor induction of Cyp1b1 in the parental strain of B6 mice, especially in fetal lung tissue. We also measured the levels of MC adducts and their disappearance from lung tissue by the P(32) post-labeling assay on gestation days 18 and 19 and postnatal days 1, 4, 11, and 18. Few differences were seen between the different strains of mice; the parental strain of B6 mice had nominally higher levels of DNA adducts 2 (gestation day 19) and 4 (postnatal day 1) days after injection, although this was not statistically significant. These results indicate that differences in Phase I metabolism of MC and formation of MC-DNA adducts are unlikely to account for the marked differences observed in the Ki-ras mutational spectrum seen in previous studies. Further, the results suggest that other genetic factors may interact with chemical carcinogens in determining individual susceptibility to these agents during development. PMID- 15885735 TI - Relationship of CD86 surface marker expression and cytotoxicity on dendritic cells exposed to chemical allergen. AB - Human peripheral blood-derived dendritic cells (DC) respond to a variety of chemical allergens by up-regulating expression of the co-stimulatory molecule CD86. It has been postulated that this measure might provide the basis for an in vitro alternative approach for the identification of skin sensitizing chemicals. We recently reported that DC, exposed in culture to the highest non-cytotoxic concentrations of various chemical allergens, displayed marginal up-regulation of membrane CD86 expression; the interpretation being that such changes were insufficiently sensitive for the purposes of hazard identification. For the work presented here, immature DC were derived from human monocytes and treated with the chemical allergens 2,4-dinitrobenzenesulfonic acid (DNBS), nickel sulfate (NiSO4), p-phenylenediamine (PPD), Bandrowski's base (BB), hydroquinone (HQ) and propyl gallate (PG) for 48 h at concentrations which induced both no to slight to moderate cytotoxicity. For comparison, DC were treated with the irritants sodium dodecyl sulfate (SDS), benzoic acid (BA), and benzalkonium chloride (BZC) at concentrations resulting in comparable levels of cytotoxicity. CD86 expression, as measured by flow cytometry, was consistently up-regulated (ranging from 162 to 386% control) on DC treated with concentrations of chemical allergens that induced approximately 10-15% cytotoxicity. The irritants BA and BZC did not induce up-regulation of CD86 expression when tested at concentrations that induced similar levels of cytotoxicity. SDS, however, up-regulated CD86 expression to 125-138% of control in 2/4 preparations when tested at concentrations which induced similar toxicity. Our results confirm that chemical allergens up-regulate CD86 expression on blood-derived DC and illustrate further that up-regulation of CD86 surface marker expression is more robust when DC are treated with concentrations of chemical allergen that induce slight to moderate cytotoxicity. PMID- 15885736 TI - Cross-neutralization of cutaneous and mucosal Papillomavirus types with anti-sera to the amino terminus of L2. AB - Vaccination with papillomavirus L2 has been shown to induce neutralizing antibodies that protect against homologous type infection and cross-neutralize a limited number of genital HPVs. Surprisingly, we found that antibodies to bovine papillomavirus (BPV1) L2 amino acids 1-88 induced similar titers of neutralizing antibodies against Human papillomavirus (HPV)16 and 18 and BPV1 pseudoviruses and also neutralized HPV11 native virions. These antibodies also neutralized each of the other pseudovirus types tested, HPV31, HPV6 and Cottontail rabbit papillomavirus (CRPV) pseudoviruses, albeit with lower titers. HPV16, HPV18, HPV31, HPV6 and CRPV L2 anti-sera also displayed some cross-neutralization, but the titers were lower and did not encompass all pseudoviruses tested. This study demonstrates the presence of broadly cross-neutralizing epitopes at the N terminus of L2 that are shared by cutaneous and mucosal types and by types that infect divergent species. BPV1 L2 was exceptionally effective at inducing cross neutralizing antibodies to these shared epitopes. PMID- 15885737 TI - Bacteria enclosure between silica-coated membranes for the degradation of organic compounds in contaminated water. AB - A non-conventional technique is proposed for the enclosure of either pure bacterial cultures or entire biocoenoses, for a possible utilization in the treatment of contaminated water. Biological components have been enclosed between polyester membranes coated by silica films consisting of: (a) SiO2 and nitrocellulose, (b) SiO2, ZnS crystals and nitrocellulose, (c) SiO2, TiO2 crystals and nitrocellulose, (d) SiO2, ZnS and TiO2 crystals and nitrocellulose. Morphological, structural and mechanical features of membranes were investigated by means of optical and electron microscopy, mercury porosimetry and wear resistance tests. Degradation kinetics have been finally studied by dipping the entrapped biomass into aqueous solutions of three different model organic compounds (alpha-d-glucose, ethyl alcohol and peptone). Results are very promising: in fact, no metabolic inhibition mechanisms of microorganisms have been evidenced. The porosity of the system allows mass transfer through the membranes, hence bacteria can grow and degrade pollutants. Besides, by this system, cells are constrained, avoiding they to spread across the retainment scaffold. PMID- 15885738 TI - Fenton-driven chemical regeneration of MTBE-spent GAC. AB - Methyl tert-butyl ether (MTBE)-spent granular activated carbon (GAC) was chemically regenerated utilizing the Fenton mechanism. Two successive GAC regeneration cycles were performed involving iterative adsorption and oxidation processes: MTBE was adsorbed to the GAC, oxidized, re-adsorbed, oxidized, and finally re-adsorbed. Oxidant solutions comprised of hydrogen peroxide (H2O2) (1.7 2.0%) and FeSO4 x 7H2O (3 g/L) (pH 2.5), were recirculated through the GAC column (30% bed expansion). The regeneration efficiency after two full cycles of treatment was calculated to be 91%. The cost of H2O2 was 0.59 dollars/kg GAC (0.27 dollars/lb) per regeneration cycle. There was no loss of sorptive capacity. Small reductions in carbon surface area and pore volume were measured. The lack of carbon deterioration under aggressive oxidative conditions was attributed to the oxidation of the target contaminants relative to the oxidation of carbon surfaces. The reaction byproducts from MTBE oxidation, tertiary butanol and acetone, were also degraded and did not accumulate significantly on the GAC. Excessive accumulation of Fe on the GAC and consequent interference with MTBE sorption and carbon regeneration was controlled by monitoring and adjusting Fe in the oxidative solution. PMID- 15885739 TI - Contaminated sediments and bioassay responses of three macroinvertebrates, the midge larva Chironomus riparius, the water louse Asellus aquaticus and the mayfly nymph Ephoron virgo. AB - Bioassays are widely used to estimate ecological risks of contaminated sediments. We compared the results of three whole sediment bioassays, using the midge larva Chironomus riparius, the water louse Asellus aquaticus, and the mayfly nymph Ephoron virgo. We used sediments from sixteen locations in the Dutch Rhine-Meuse Delta that differed in level of contamination. Previously developed protocols for each bioassay were followed, which differed in sediment pretreatment, replication, and food availability. The Chironomus bioassay was conducted in situ, whereas the other two were conducted in the laboratory. The measured endpoints, survival and growth, were related to contaminant levels in the sediment and to food quantity in water and sediment. Only the response of A. aquaticus in the bioassay was correlated with sediment contamination. Food availability in overlying water was much more important for C. riparius and E. virgo, thereby masking potential sediment contaminant effects. We conclude that growth of A. aquaticus was depressed by sediment contamination, whereas growth of E. virgo and C. riparius was stimulated by seston food quantity. We discuss that the trophic state of the ecosystem largely affects the ecological risks of contaminated sediments. PMID- 15885740 TI - Radiolytic degradation of Acid Orange 7: a mechanistic study. AB - Steady-state radiolysis experiments were performed to investigate the mechanisms of the radiolytic degradation of Acid Orange 7 (AO7) in aqueous solutions, which might be useful for the application of ionizing radiation for the remediation of azo-dye-laden wastewaters. The degradation products formed under various conditions were identified by using UV-Vis, HPLC, FTIR, and GC-MS analyses. With theoretical analysis and degradation products identified, the mechanisms behind the radiolytic degradation of AO7 under both oxidative and reductive conditions were elucidated. Irradiated under reductive conditions AO7 was decomposed through N-N cleavage with the formation of aniline, sodium sulfanilamide, 1-amino-2 naphthol, naphthalidine, 1,2,3,4-tetrahydro-2-naphthol, and 2-naphthol etc., whereas under oxidative conditions both N-N and C-N cleavages might be the initial steps in the radiolytic degradation of AO7. PMID- 15885741 TI - Extractable organic halogens (EOX) in sediments from selected Polish rivers and lakes--a measure of the quality of the inland water environment. AB - EOX in sediments from the two main Polish rivers (the Vistula and the Oder/Odra) as well as five lakes was assayed coulometrically and its values examined with respect to the location of the sample collection points. EOX values were found to increase near industrial areas and large urban agglomerations, although mean values--comparable for rivers and lakes--were similar to those recorded elsewhere. EOX tends to rise as TOC does so. On the other hand, the EOX:TOC ratio decreases sharply in the case of lakes, since TOC values there are roughly one order of magnitude higher than in rivers. Furthermore, the contents of selected polychlorinated biphenyls and organochlorine pesticides are directly related to EOX values. These relationships constitute a basis for the application of EOX as a parameter reflecting the level of organochlorine compounds in environmental quality evaluation. PMID- 15885742 TI - The endocrine and reproductive function of the female Yucheng adolescents prenatally exposed to PCBs/PCDFs. AB - BACKGROUND: Polychlorinated biphenyls (PCBs) and related compounds such as polychlorinated dibenzofurans (PCDFs) and polychlorinated dibenzo-dioxins (PCDDs) alter sexual maturation and endocrine function in animals. In 1978-1979, a mass poisoning occurred in central Taiwan from cooking oil contaminated by heat degraded PCBs and oxidated compounds PCDFs. We tested the hypothesis that in utero exposed to PCBs/PCDFs alter sexual maturation, endocrine, and reproductive function in the human pubescent females. METHODS: In 1997-1999, girls aged 13-19 years, born to mothers exposed to PCBs/PCDFs, was invited to participate in the study. Menstruation characteristic was recorded daily for 84 days and serum levels of estradiol, LH, FSH, and testosterone were measured on the 3rd day of menstruation. RESULTS: A total of 17 exposed girls and controls participated, the exposed girls reported shorter mean duration of bleeding per cycle than 16 unexposed (5.5 vs 6.5 days, P=0.0055). There was a higher rate of irregular menstrual cycle in the exposed girls (40% vs 0%, P=0.018). Serum levels of estradiol (P=0.016) and FSH (P=0.061) were higher in exposed girls as compared to controls. CONCLUSIONS: We conclude that prenatal exposure to PCBs/PCDFs resulted in abnormal menstruation and higher estradiol and FSH levels in follicular phase of menstrual cycle in adolescent girls. PMID- 15885743 TI - Uptake and degradation of DDT by hairy root cultures of Cichorium intybus and Brassica juncea. AB - Hairy root cultures of Cichorium intybus and Brassica juncea were used for their ability to uptake and degrade DDT (1,1,1-trichloro-2,2-bis-(4' chlorophenyl)ethane). After 24 h of 14C DDT treatment, only 12-13% of the total applied radioactivity was detected in the culture media, indicating the efficient uptake of DDT by the hairy roots. The majority of the applied radioactivity was associated with the roots. DDT was degraded to various other products such as DDD, DDE and DDMU, along with some unknown compounds by hairy root cultures, which were detected by thin layer chromatography (TLC) and autoradiography. The rate of in situ degradation was found to be higher during the initial stages of culture and the residual 14C DDT in the roots was found to decrease from 77% to 61% over a period of 10-days. There was no spontaneous degradation of 14C DDT in media lacking hairy root cultures or in media with autoclaved hairy roots. This suggests that endogenous root enzymes play a role in the breakdown of 14C DDT. These results suggest the potential applicability and advantage of using these plant species for phytoremediation of persistent xenobiotics such as DDT in an eco-friendly and efficient manner for environmental clean up. PMID- 15885745 TI - Overview of caloric restriction and ageing. AB - It has been known for some 70 years that restricting the food intake of laboratory rats extends their mean and maximum life span. In addition, such life extension has been observed over the years in many other species, including mice, hamsters, dogs, fish, invertebrate animals, and yeast. Since this life-extending action appears to be due to a restricted intake of energy, this dietary manipulation is referred to as caloric restriction (CR). CR extends life by slowing and/or delaying the ageing processes. The underlying biological mechanism responsible for the life extension is still not known, although many hypotheses have been proposed. The Growth Retardation Hypothesis, the first proposed, has been tested and found wanting. Although there is strong evidence against the Reduction of Body Fat Hypothesis, efforts have recently been made to resurrect it. While the Reduction of Metabolic Rate Hypothesis is not supported by experimental findings, it nevertheless still has advocates. Currently, the most popular concept is the Oxidative Damage Attenuation Hypothesis; the results of several studies provide support for this hypothesis, while those of other studies do not. The Altered Glucose-Insulin System Hypothesis and the Alteration of the Growth Hormone-IGF-1 Axis Hypothesis have been gaining favor, and data have emerged that link these two hypotheses as one. Thus, it may now be more appropriate to refer to them as the Attenuation of Insulin-Like Signaling Hypothesis. Finally, the Hormesis Hypothesis may provide an overarching concept that embraces several of the other hypotheses as merely specific examples of hormetic processes. For example, the Oxidative Damage Attenuation Hypothesis probably addresses only one of likely many damaging processes that underlie aging. It is proposed that low-intensity stressors, such as CR, activate ancient hormetic defense mechanisms in organisms ranging from yeast to mammals, defending them against a variety of adversities and, when long-term, retarding senescent processes. PMID- 15885746 TI - Scavenging of gaseous mercury by acidic snow at Kuujjuarapik, Northern Quebec. AB - One fate of gaseous elemental mercury (GEM) in the Arctic has been identified as gas phase oxidation by halogen-containing radicals, leading to abrupt atmospheric mercury depletion concurrent with ozone depletion. Rapid deposition of oxidized mercury leads to snow enrichment in mercury. In this report, we describe experiments that demonstrate the ability of snow to directly scavenge atmospheric mercury. The study was conducted at Kuujjuarapik, Quebec, Canada (latitude 55 degrees 17'N). A mercury depletion event (MDE) caused the mercury concentration in the surface snow of the coastal snowpack to double, from (9.4+/-2.0) to (19.2+/-1.7) ng/L. Independent of the MDE, mercury concentrations increased five fold, from (10.0+/-0.1) to (51.4+/-6.0) ng/L, upon spiking the snow with 500 microM hydrogen peroxide under solar irradiation. Total organic carbon in the spiked irradiated snow samples also decreased, consistent with the formation of strongly oxidizing species. The role of the snowpack in releasing GEM to the atmosphere has been reported; these findings suggest that snow may also play a role in enhancing deposition of mercury. PMID- 15885747 TI - Blood lead levels in lactating cows reared around polluted localities; transfer of lead into milk. AB - Lead is pervasive environmental pollutant with potential public health hazard as a contaminant of food from animal origin. The present study examines the blood and milk lead level in animals reared in areas around different industrial activities and to find out correlation between blood and milk lead levels in lactating cows. Blood and milk samples (n = 149) were collected from animals reared around steel processing unit (n = 22), lead-zinc smelter (n = 21), aluminum processing plant (n = 25), rock phosphate mining area cum phosphate fertilizer plant (n = 21), coal mining areas (n = 46) and closed lead but functional zinc smelter (n = 14). Samples were also collected from randomly chosen 52 lactating cows reared in non-polluted areas to serve as controls. Significantly (P < 0.05) higher blood lead level was recorded in animals reared around lead-zinc smelting factories followed by closed lead but functional zinc smelter, aluminum processing unit and steel manufacturing plant, as compared to values recorded for control animals. The highest milk lead level (0.84 +/- 0.11 microg/ ml) was detected in animals reared in the vicinity of lead-zinc smelting unit followed by aluminum processing plant and steel processing unit. Analysis of correlation between blood lead levels and lead excretion in milk through sorting the blood lead values into 9 different ranges irrespective of site of collection of samples (n = 201) revealed significant correlation (r = 0.469 at P < 0.01) between blood and milk lead concentrations. The lactating cows with blood lead levels above 0.20 microg/ml (groups 5-9) had significantly (P < 0.05) higher milk lead excretion than those with blood lead levels from non detectable to 0.20 microg/ml (groups 1-4). Pearson correlation analysis between blood and milk lead concentrations in 122 animals with blood lead <0.20 microg/ml showed non significant correlation (r = 0.030 at P < 0.05) but a significant correlation was observed between these two parameters with blood lead level above >0.02 microg/ml indicating that the excretion of lead through milk increases with the increased in blood lead level above 0.20 microg/ml. PMID- 15885748 TI - Multivariate analysis of heavy metal contamination in urban dusts of Xi'an, Central China. AB - Though there are many studies of heavy metal contaminations of urban dusts in developed countries, little attention has been paid to this kind of study in developing countries, including China. Therefore, a series of investigations were performed to provide heavy metal signatures of urban dusts and to evaluate potential sources in Xi'an, Shaanxi Province. Sixty-five samples of urban dusts were collected in Xi'an. Then Ag, Cr, Cu, Mn, Pb and Zn concentrations were determined by using atomic absorption spectrophotometry, and As, Hg and Sb concentrations by atomic fluorescence spectroscopy. The results indicate that, in comparison with Chinese soil, urban dusts in Xi'an have elevated metal concentrations as a whole, except those of arsenic and manganese. These concentration levels are comparable to those in other studies. Correlation coefficient analysis, principal component analysis (PCA) and cluster analysis (CA) were performed and three main sources with corresponding cluster elements were identified: (1) Ag and Hg have commercial and domestic sources; (2) Cr, Cu, Pb, Sb and Zn are mainly derived from industrial sources, combined with traffic sources as well for Pb and Zn; (3) As and Mn come mainly from soil sources, and As also has an industrial source. Based on PCA and CA analyses, manganese was selected as the reference element, and heavy metal enrichment factors (Efs) were calculated, which in turn further confirms the source identification. Also, Efs give an insight of human influence degree of urban dusts. PMID- 15885749 TI - A long-term increase in eggshell thickness of Greenlandic Peregrine Falcons Falco peregrinus tundrius. AB - Thickness of eggshell fragments and whole eggs from the Peregrine Falcon Falco peregrinus collected in South and West Greenland between 1972 and 2003 was measured and compared to shell thickness of pre-DDT eggs, also collected in Greenland. Linear regression yields a significant increase in the average thickness of eggshells over the period of 0.19% per year, corresponding to a change in eggshell thinning from 13.9% in 1972 to 7.8% in 2003. Backwards extrapolation of the data, suggests that the Greenlandic Peregrine population probably was never critically affected by DDT-induced eggshell thinning. By sampling eggshell fragments in many nests the spatial and temporal sample distribution was enlarged, allowing the detection of a significant long-term decrease in pollutant-induced eggshell thinning--a trend that could not have been identified if only the rarer whole, addled eggs had been sampled. PMID- 15885750 TI - Dissolved natural organic matter (NOM) impacts photosynthetic oxygen production and electron transport in coontail Ceratophyllum demersum. AB - Dissolved natural organic matter (NOM) is dead organic matter exceeding, in freshwater systems, the concentration of organic carbon in all living organisms by far. 80-90% (w/w) of the NOM is made up of humic substances (HS). Although NOM possesses several functional groups, a potential effect on aquatic organisms has not been studied. In this study, direct effects of NOM from various origins on physiological and biochemical functions in the aquatic plant Ceratophyllum demersum are presented. Environmentally relevant concentrations of NOM cause inhibitory effects on the photosynthetic oxygen production of C. demersum. Various NOM sources and the synthetic humic substance HS1500 inhibit the photosynthetic oxygen production of the plant as observed with 1-amino anthraquinone, a known inhibitor of plant photosynthesis. 1-Aminoanthraquinone may serve as an analogue for the quinoid structures in NOM and HS. Most likely, the effects of NOM may be related to quinoid structures and work downstream of photosynthesis at photosystem (PS) II. PMID- 15885751 TI - Distribution of polycyclic aromatic hydrocarbons in riverine waters after Mediterranean forest fires. AB - Extensive forest fires occurred in Catalonia, northern Spain, in 1994. In our study, concentrations and profiles of 12 parent polycyclic aromatic hydrocarbons (PAHs) were determined in riverine waters, ash and sediment samples at nine sampling sites (W1-W9) and at three sampling dates from Llobregat hydrographic basin: in August, 1994, one month after the extensive forest fires; in September, 1994, after the first heavy autumn rainfalls and in January, 1995, six months after forest fires. In August 1994, the total concentrations of 12 PAHs measured in riverine waters varied from 2 ng/l to 336 ng/l. In September 1994, the total PAH concentrations decreased to 0.2-31 ng/l and in January 1995, from 9 ng/l to 73 ng/l. In August, the composition pattern of PAHs showed a distribution dominated by 4-ring PAHs (pyrene, chrysene+triphenylene, benzo(a)anthracene) at W3-W6, W8 and W9 and 3-ring PAHs (phenanthrene) at W1, W2 and W7. In September, a preference by 3-ring PAHs (phenanthrene) at all sampling sites except W5 was shown and in January was clearly dominated by 4-ring PAHs (chrysene+triphenylene, pyrene, benzo(a)anthracene) at all sampling sites. In ash and sediment samples, the total concentrations of 12 PAHs ranged from 1.3 ng/g to 19 ng/g. The dominant compound was phenanthrene. PMID- 15885752 TI - Exposure assessment of organic solvents for aircraft paint stripping and spraying workers. AB - The main objective of this study is to investigate the personal or area exposure of organic solvents during paint stripping and paint spraying. Three aircraft paint stripping/spraying workplaces in Taiwan were selected, and the Council of Labor Affairs and NIOSH recommended sampling/analytical methods used in this study. Activated charcoal tubes were used to investigate the personal and area exposure concentration of organic solvents in paint stripping and paint spraying operations. During aircraft paint stripping, experiment results show that methylene chloride personal exposure concentration at the ground area, 42.01+/ 31.86 ppm, is higher than that at the working platform 4 M high above the ground, 20.41+/-11.43 ppm. Exposure concentration of methylene chloride in the initial paint stripping operation stage of every workplace is over the PEL (50 ppm) set by the Taiwan Council of Labor Affairs. Corrective actions are needed. During paint spraying, concentrations of all organic solvents were found to be below the PEL of OSHA. PMID- 15885753 TI - Distribution of brominated flame retardants in different dust fractions in air from an electronics recycling facility. AB - Twelve air samples were collected from an electronic recycling facility in Sweden representing three different dust fractions; respirable, total and inhalable dust. Four samples were collected from each fraction. The highest concentration of polybrominated diphenyl ether (PBDE) #209 (ten bromine atoms) was found in the samples from the inhalable dust fraction (ID), which was 10 times higher than for the "total dust" fraction (TD). The concentration ranges were 157.6-208.6; 13.9 16.7; and 2.8-3.3 ng/m3 for inhalable, total and respirable fractions, respectively. The second most abundant PBDE congener was PBDE #183 (seven bromine atoms), followed by the second most abundant substance 1,2-bis(2,4,6 tribromophenoxy)ethane (BTBPE) in all samples. In addition, decabromodiphenyl ethane (DeBDethane) was tentatively identified in five of the samples. Because of the large differences in air concentrations between the three fractions in ID, TD and RD, it is suggested that the inhalable instead of "total dust" fraction should be used to assess air concentrations, in particular for the larger and higher brominated flame retardants (BFRs). PMID- 15885754 TI - Accumulation of butyltin compounds in cobia Rachycentron canadum raised in offshore aquaculture sites. AB - Butyltin residues (monobutyltin, MBT; dibutyltin, DBT; tributyltin, TBT; tetrabutyltin, TeBT) in the sea water and in the cobia (Rachycentron canadum) from aquaculture sites located offshore of Penhu island, Taiwan, were collected and quantified. The average concentrations of MBT, DBT, TBT and TeBT in sea water were n.d.-28+/-3, 4.0+/-0.6-88+/-13, n.d.-43+/-4, and n.d.-7+/-1 ng l(-1), respectively. The total butyltin (sum of MBT, DBT, TBT, TeBT) residues in the skin, dorsal muscle, ventral muscle, dark muscle, and liver of the cobia were in the range of 72+/-12-2270+/-85, 79+/-11-688+/-33, 82+/-14-1715+/-104, 93+/-13 803+/-47, and n.d.-52,745+/-252 ng g(-1) (wet weight), respectively. Although in this study in most cases, the highest concentration of total butyltin residues was found in liver or skin, in some cases, the highest concentration was found in muscle tissue. The crude lipid content in the skin, dorsal muscle, ventral muscle, dark muscle, and liver of these cobia was in the range of 7.9+/-0.1-28+/ 1%, 11.7+/-0.8-29+/-1%, 11.5+/-0.3-44+/-3%, 24.2+/-0.4-48.4+/-0.4%, and 55.7+/ 0.1-87.7+/-0.4% (wet weight), respectively. The concentrations of crude lipid content, and the concentrations of total butyltin residues in these tissues were not correlated. PMID- 15885755 TI - Theoretical stability assessment of uranyl phosphates and apatites: selection of amendments for in situ remediation of uranium. AB - Addition of an amendment or reagent to soil/sediment is a technique that can decrease mobility and reduce bioavailability of uranium (U) and other heavy metals in the contaminated site. According to data from literature and results obtained in field studies, the general mineral class of apatites was selected as a most promising amendment for in situ immobilization/remediation of U. In this work we presented theoretical assessment of stability of U(VI) in four apatite systems (hydroxyapatite (HAP), North Carolina Apatite (NCA), Lisina Apatite (LA), and Apatite II) in order to determine an optimal apatite soil amendment which could be used for in situ remediation of uranium. In this analysis we used a theoretical criterion which is based on calculation of the ion-ion interaction potential, representing the main term of the cohesive energy of the matrix/pollutant system. The presented results of this analysis indicate (i) that the mechanism of immobilization of U by natural apatites depends on their chemical composition and (ii) that all analyzed apatites represent, from the point of view of stability, promising materials which could be used in field remediation of U-contaminated sites. PMID- 15885756 TI - Influence of temporal variations in water chemistry on the Pb isotopic composition of rainbow trout (Oncorhynchus mykiss). AB - Field and laboratory investigations were undertaken to determine (1) the relations between discharge, Pb concentration, and the Pb isotopic composition of the dissolved load in Richland Creek, western North Carolina, and (2) the potential influence of varying Pb water chemistry on the Pb isotopic abundances in liver and bone tissues of rainbow trout (Oncorhynchus mykiss). Stream waters were characterized by relatively low Pb concentrations during periods of base flow exceeding 10 days in length. Moreover, greater than 65% of the Pb was derived from orchard soils located upstream of the monitoring site which are contaminated by lead arsenate. During small to moderate floods, the dissolved load exhibited Pb concentrations more than twice as high as those measured during base flow, but the contribution of Pb from lead arsenate was relatively low and varied directly with discharge. In contrast to smaller events, Pb from lead arsenate in an 8- to 10-year (overbank) event in May 2003 was minimal during peak flow conditions, suggesting that discharge-source relations are dependent on flood magnitude. The hydrologic and geochemical data demonstrate that aquatic biota in Richland Creek are subjected to short-term variations in Pb concentrations and Pb isotopic abundances within the dissolved load ranging from a few hours to few a weeks. Laboratory studies demonstrated that when rainbow trout were exposed to elevated Pb concentrations with a distinct isotopic fingerprint, the bone and liver rapidly acquire isotopic ratios similar to that of the water. Following exposure, bone retains Pb from the contaminant source for a period of months, while the liver excreted approximately 50% of the accumulated Pb within a few days and nearly all of the Pb within a few weeks. Differences in the rates of excretion resulted in contrasting isotopic ratios between the tissues. It seems plausible, then, that previously observed differences between the isotopic composition of bone and liver in rainbow trout from Richland Creek are related to their short-term exposure to Pb from soils contaminated with Pb arsenate that result from fluctuating hydrologic conditions. If these trends prove to be common in other contaminated aquatic environments, it may be possible to use the Pb isotopic composition of bone as an indicator of the long-term exposure to Pb and the liver as a biomarker for short-term Pb exposures. PMID- 15885757 TI - Dietary diacylglycerol extenuates arterial thrombosis in apoE and LDLR deficient mice. AB - INTRODUCTION: Prevention of arterial thrombotic diseases has high priority in developed countries. An appropriate diet is thought to be the best way to prevent or reduce the risk of mortality from such diseases. The aim of the present study was to evaluate the effect of diacylglycerol (DAG)-rich diets on arterial thrombosis. MATERIAL AND METHODS: Diet-sensitive congenital apolipoprotein E (ApoE) and LDL receptor (LDLR) double deficient mice were used. Thrombosis was assessed by the rate and extent of thrombus formation in the carotid artery of mice after laser irradiation. Plasma total cholesterol and triglyceride levels were measured by enzymatic assays. Four kinds of diets were used: high fat (Western-style) diet contained 20% fat (w/w) and 0.05% cholesterol (w/w); the TAG rich and the DAG-rich high fat diet contained 20% TAG or DAG oil (w/w) with very similar fatty acid composition and 0.05% cholesterol ; Low fat (Japanese-style) diet contained 7% fat, without cholesterol. These diets were on the basis of AIN93G and were given to mice for 8 weeks from 6 weeks of age. RESULTS: Western style high fat and TAG-rich high fat diets significantly increased thrombogenicity compared with low fat diet. DAG-rich high fat diet showed the lowest value, and the extent of thrombogenicity was equivalent to the low fat diet group. Fasting plasma total cholesterol level of DAG-rich high fat and low fat diet groups were significantly lower than that of TAG-rich high fat and high fat diet groups. Fasting plasma triglyceride levels in DAG-rich high fat diet group were significantly lower than in the TAG-rich high fat diet group. CONCLUSIONS: Dietary DAG but not TAG oil extenuates arterial thrombus formation. The mechanism of this effect is unclear and further investigated. PMID- 15885758 TI - No increase in CD62P-positive single platelets after acute platelet activation in vivo. PMID- 15885759 TI - Ionic liquids as an alternative to formalin in histopathological diagnosis. AB - Asymmetry of cations and the type of anions play a key role in the properties of ionic liquids (ILs) as fixatives for tissue preservation. 1-Methyl-3 octyloxymethylimidazolium tetrafluoroborate has proven to be a very good fixative, with similar effects as formalin. Our study shows that it is applicable for both histological and immunohistochemical purposes. After treatment with 1 methyl-3-octyloxymethylimidazolium tetrafluoroborate, tissue sections are more intensely stained. With respect to expression patterns and staining intensity, immunohistochemical staining is comparable in tissues fixed in formalin and the selected ILs. The present study demonstrates the properties of 1-methyl-3 octyloxymethylimidazolium tetrafluoroborate for tissue preservation in histopathological procedures, eliminating the requirement of formalin. PMID- 15885760 TI - Merkel cell carcinoma of the Bartholin's gland. AB - BACKGROUND: There are a few cases of "small cell neuroendocrine carcinoma" of the Bartholin's gland as well as Merkel cell carcinoma of the vulva reported in the gynecologic oncology literature, most with aggressive clinical behavior. Merkel cells were originally described in the epidermis with neuroendocrine properties. Merkel cell carcinoma is different from the neuroendocrine small cell (oat cell) tumor arising in the lung and other internal organs based on morphology and immunohistochemistry. CASE: A 49-year-old female was diagnosed with a FIGO stage I Merkel cell carcinoma of the Bartholin's gland. The tumor originated from the duct and stained with endocrine markers and cytokeratin 7 and 20. She was treated with radical wide local excision and bilateral inguinal lymph node dissection followed by radiation therapy to the pelvis, perineum, vulva, and inguinal regions. Two years after the diagnosis, she is alive with no evidence of recurrent disease. CONCLUSION: This is a case of Merkel cell carcinoma of the Bartholin's gland discovered in the early stage with a survival of 2 years following diagnosis and treatment with no evidence of disease recurrence. PMID- 15885761 TI - Bcl-2, BAX, and apoptosis in endometrial hyperplasia after high dose gestagen therapy: a comparison of responses in patients treated with intrauterine levonorgestrel and systemic medroxyprogesterone. AB - OBJECTIVES: The aim of the study was to investigate apoptosis as a growth regulatory mechanism of gestagen in endometrial precancers and to compare differences in the apoptotic cascade after high and low dose gestagen regimens. METHOD: Pre- and post-treatment paraffin-embedded endometrial hyperplasia specimens from women treated with levonorgestrel intrauterine device (n = 26) and women treated with 10 mg medroxyprogesterone for 10 days per cycle (n = 31) were examined for changes in the expression of Bcl-2 and BAX and the extent of apoptosis after 3 months of treatment. Immunohistochemical expression in tissue specimens for Bcl-2 and BAX was evaluated by H-score. Average number of apoptotic cells per hundred cells within ten different high power field (40 x) was evaluated for each section after in situ apoptosis detection (TUNEL method). A second group of patients with endometrial hyperplasia was examined after 1 week treatment with levonorgestrel IUD (n = 6) or medroxyprogesterone (n = 5) to determine early effects on expression of Bcl-2 and BAX and the extent of apoptosis. RESULTS: All the patients in the IUD group (n = 31) but only about half of the patients in per oral group (16 of 26) responded to treatment. The glandular reduction in Bcl-2 expression was markedly greater for the IUD patients than for the patients who received oral gestagen. The decrease in BAX expression after IUD treatment was less than the reduction of Bcl-2. Decrease in glandular Bcl-2 after 3 months of treatment was coincident with a significant increase in the measurable amount of apoptosis. In stromal cells, the increase in expression of Bcl-2 and BAX was found after gestagen treatment, the response being much more marked for the IUD group. The non- responders to per oral gestagen had no Bcl-2 expression in stroma after 3 months of therapy whereas an increase was observed for the responders. After 1 week, glandular Bcl-2 expression was significantly reduced after treatment in the IUD group. As for the rest, no changes were detected in either of the groups. CONCLUSION: Our results indicate that proteins in the apoptotic cascade are regulated by gestagen therapy in human endometrial precancers. Expression of these proteins is shown to be dependent on administration form and/or type of gestagen. Stromal Bcl-2 expression appears to be a potential biomarker which can separate responders of gestagen treatment from non-responders after oral administration. PMID- 15885762 TI - Non-puerperal uterine inversion in association with uterine sarcoma: case report in a 26-year-old and review of the literature. AB - INTRODUCTION: Inversion of the uterus associated with a uterine sarcoma is extremely rare with only 17 cases reported since 1887, so that its clinical presentation and appearance at laparotomy may be confusing. CASE: A 26-year-old woman presented with a vaginal mass and bleeding. At laparotomy, her uterus appeared depressed or concave. The final diagnosis of uterine sarcoma and inversion was unique in her age group. CONCLUSIONS: Uterine sarcoma with inversion can occur in the premenopausal age group. It presents as a vaginal mass with bleeding. As a clue to the diagnosis, the uterus per se may not be palpable or it may appear concave on physical exam or at laparotomy. PMID- 15885763 TI - Proviral DNA of caprine arthritis encephalitis virus (CAEV) is detected in cumulus oophorus cells but not in oocytes from naturally infected goats. AB - The aim of this study was to determine whether oocytes taken from ovarian follicles in 123 naturally infected goats were carrying the proviral CAEV genome. Examination of DNA isolated from 190 batches of oocytes with intact cumulus cells and 190 batches of oocytes whose cumulus cells had been removed, taken from follicles of the same ovaries, demonstrated that 42/190 batches of oocytes with intact cumulus cells had the proviral CAEV genome, whereas none of the 190 batches of oocytes without cumulus cells were positive for the provirus. To confirm that the proviral genome was present in the cumulus cells and not in the oocyte cells, 586 oocytes from 56 different ovaries, were separated from their cumulus cells. The DNA was then extracted from each fraction and examined. The purity of the oocyte fraction was verified by searching for granulosa cell specific mRNA, using RT-PCR; this was negative in all the batches of oocytes in which the cumulus cells had been removed. PCR analysis demonstrated that none of the oocytes without cumulus cells were positive, whereas 22/56 of the batches with cumulus cells were found to be positive. This study clearly demonstrates that despite being surrounded by infected cumulus cells, the oocytes are not infected, and that the enzymatic and mechanical technique for removing the cells surrounding the oocyte, as used in this study, is effective, thus enabling CAEV free oocytes to be obtained from infected goats. PMID- 15885764 TI - How filler properties, filler fraction, sample thickness and light source affect light attenuation in particulate filled resin composites. AB - OBJECTIVE: It was hypothesized that by standardizing variables such as light sources, filler types and filler surface treatment, it should be possible to use Beer-Lambert's law to predict light absorption in visible light-cured dental composites. METHODS: Mixture of 50 wt% bisGMA and 50 wt% TEGDMA to which a photo initiator (0.35 wt% champhorquinone) and a co-initiator (0.7 wt% of dimethylaminoethylmethacrylate) was prepared. Three different filler types (HBB, SBB and KU) were added to that mixture in eight different volume percentage. Filler particles were either silane surface treated or not. Specimens were made with thicknesses of 1-5 mm. Total number of 1200 specimens were made for this study. Light transmission was obtained for halogen source and laser lights, which made the number of observations 2400. The absorbance values of the different materials were analyzed in Matlab with respect to the differences in filler fraction and sample thickness. RESULTS: The obtained results revealed that of the two light sources, more light was absorbed by the composite when the laser light was used. Among different filler types, the HBB filler absorbed most light and the KU filler the least. There were significant differences (p < 0.05) in light absorption between all three filler types. SIGNIFICANCE: By comparing the modeled surfaces generated by Matlab for different materials it was possible to determine how different variables such as filler type, filler surface treatment and light source affect light attenuation. The characteristic of incident light affected the light absorbance, meaning that not only the composite's composition needs to be considered in light absorption studies of dental composites. PMID- 15885765 TI - Effect of water storage, thermocycling, the incorporation and site of placement of glass-fibers on the flexural strength of veneering composite. AB - OBJECTIVES: To evaluate the effects of water storage, thermocycling, and the incorporation of glass-fibers, on the flexural strength of veneering composites. METHODS: Veneering composites with different fillers, matrices and polymerization methods (Belleglass Kerr Inc., Orange, CA, USA; Sculpture, Pentron Inc. Wallingford CT, USA; Sinfony, 3M Espe, Seefeld, Germany; SR Adoro LC and HP, Targis, Ivoclar Vivadent, Schaan, Liechtenstein), a glass-fiber framework material (Vectris Pontic VP, Ivoclar Vivadent) and a direct restorative composite control (Tetric Ceram, Ivoclar Vivadent) were selected. For the first part of the study, 30 bar specimens (25 x 2 x 2 mm3) per material were fabricated. Ten were stored for 24 h and 10 for 14 days in water at 37 degrees C. Ten were thermocycled (3000 x; 5-50-5 degrees C). Three-point bending tests (crosshead speed: 0.5 mm/min) were performed. For the second part of the study, all veneering materials were combined with a glass-fiber framework (VP). Sixty specimens were produced for each material (25 x 4 x 2 mm3) and treated as in the first part. Three-point bend tests were performed with the reinforcing glass fiber framework either on the tension or the compression side. Data was evaluated by ANOVA and Weibull analysis. RESULTS: A decrease in flexural strength was observed after water storage or thermocycling for all veneering materials tested. None of the tested materials exhibited significant advantages compared to the control. The flexural strength of glass-fiber reinforced frameworks was ten times higher and not influenced by water storage or thermocycling. A significant reinforcing effect from glass fibers was observed when they were placed on the tension but not when placed on the compression side. SIGNIFICANCE: A glass-fiber framework on the tension side significantly improved the flexural strength of veneering composites. There was less deterioration due to water storage and thermocycling with the glass-fiber reinforced veneering composite compared to the non-reinforced materials. PMID- 15885766 TI - Chitosan/beta-lactoglobulin core-shell nanoparticles as nutraceutical carriers. AB - Chitosan (CS)/beta-lactoglobulin (betalg) core-shell nanoparticles (CS-betalg nanoparticle) were successfully prepared with the aim of developing a biocompatible carrier for the oral administration of nutraceuticals. The effects of pH and initial concentrations (C(beta)(lg)) of native and denatured betalg on the properties of the nanoparticles were investigated. Uniform nanoparticles were prepared by ionic gelation with sodium tripolyphosphate (TPP). The surface charge of the particles was positive, with a zeta potential of 20-60 mV. (beta)lg loading efficiency (LE) spanned a broad range (1-60%); and was highly sensitive to formulation pH. This adsorption can be mainly attributed to electrostatic, hydrophobic interactions and hydrogen bonding between (beta)lg and CS. Brilliant blue (BB) release experiments showed that the nanoparticles prepared with native (beta)lg had favorable properties to resist acid and pepsin degradation in simulated gastric conditions unlike those prepared with denatured (beta)lg or denatured (beta)lg crosslinked with Ca2+. When transferred to simulated intestinal conditions, the (beta)lg shells of the nanoparticles were degraded by pancreatin. PMID- 15885767 TI - A histological evaluation for guided bone regeneration induced by a collagenous membrane. AB - This study was designed to evaluate the histological changes during ossification and cellular events including osteogenic differentiation responding to collagenous bioresorbable membranes utilized for GBR. Standardized artificial bony defects were prepared at rat maxillae, and covered with a collagenous bioresorbable membrane. These animals were sacrificed at 1, 2, 3 and 4 weeks after the GBR-operation. The paraffin sections were subject to tartrate resistant acid phosphatase (TRAP) enzyme histochemistry and immunohistochemistry for alkaline phosphatase (ALP), osteopontin (OP) and osteocalcin (OC). In the first week of the experimental group, woven bone with ALP-positive osteoblasts occupied the lower half of the cavity. The collagenous membrane included numerous ALP negative cells and OP-immunoreactive extracellular matrices. At 2 weeks, the ALP , OP- and OC-immunoreactivity came to be recognizable in the region of collagenous membrane. Since ALP-negative soft tissue separated the collagenous membrane and the new bone originating from the cavity bottom, the collagenous membrane appeared to induce osteogenesis in situ. At 3 weeks, numerous collagen fibers of the membrane were embedded in the adjacent bone matrix. At 4 weeks, the membrane-associated and the cavity-derived bones had completely integrated, showing the same height of the periosteal ridge as the surrounding alveolar bones. The collagen fibers of a GBR-membrane appear to participate in osteogenic differentiation. PMID- 15885768 TI - Improvement of surface bioactivity on titanium by water and hydrogen plasma immersion ion implantation. AB - We have investigated the surface bioactivity of titanium after water and hydrogen plasma immersion ion implantation. Plasma immersion ion implantation (PIII) excels in the surface treatment of components possessing a complicated shape such as medical implants. In addition, water and hydrogen PIII has been extensively studied as a method to fabricate silicon-on-insulator (SOI) substrates in the semiconductor industry and so it is relatively straightforward to transfer the technology to the biomedical field. In our investigation, water and hydrogen were plasma-implanted into titanium sequentially. Our objective is that water PIII introduces near-surface damages that trap hydrogen implanted in the subsequent step to improve the surface bioactivity while the desirable bulk properties of the materials are not compromised. Ti-OH functional groups can be detected on the (H(2)O+H(2))-implanted titanium surface by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. After incubation in simulated body fluids (SBF) for cytocompatibililty evaluation in vitro, bone-like hydroxyapatite was found to precipitate on the (H(2)O+H(2)) implanted samples while no apatite was found on titanium samples plasma implanted with water or hydrogen alone. Human osteoblast cells were cultured on the (H(2)O+H(2)) implanted titanium surface and they exhibited good adhesion and growth. Our results suggest a practical means to improve the surface bioactivity and cytocompatibility of medical implants made of titanium. PMID- 15885769 TI - Mechanical properties and osteoconductivity of porous bioactive titanium. AB - Porous bioactive titanium implants (porosity of 40%) were produced by a plasma spray method and subsequent chemical and thermal treatments of immersion in a 5M aqueous NaOH solution at 60 degrees C for 24 h, immersion in distilled water at 40 degrees C for 48 h, and heating to 600 degrees C for 1 h. Compression strength and bending strength were 280 MPa (0.2% offset yield strength 85.2 MPa) and 101 MPa, respectively. For in vivo analysis, bioactive and nontreated porous titanium cylinders were implanted into 6mm diameter holes in rabbit femoral condyles. The percentage of bone-implant contact (affinity index) of the bioactive implants (BGs) was significantly larger than for the nontreated implants (CGs) at all postimplantation times (13.5 versus 10.5, 16.7 versus 12.7, 17.7 versus 10.2, 19.1 versus 7.8 at 2, 4, 8 and 16 weeks, respectively). The percentage of bone area ingrowth showed a significant increase with the BGs, whereas with the CGs it appeared to decrease after 4 weeks (10.7 versus 9.9, 12.3 versus 13.1, 15.2 versus 9.8, 20.6 versus 8.7 at 2, 4, 8 and 16 weeks, respectively). These results suggest that porous bioactive titanium has sufficient mechanical properties and biocompatibility for clinical use under load-bearing conditions. PMID- 15885770 TI - Electrospun chitosan-based nanofibers and their cellular compatibility. AB - Chitosan-based nanofibers with an average fiber diameter controllable from a few microns down to approximately 40 nm and a narrow size distribution were fabricated by electrospinning solutions containing chitosan, polyethylene oxide (PEO), and Triton X-100. Rheological study showed a strong dependence of spinnability and fiber morphology on solution viscosity and thus on chitosan-to PEO ratio. The nanofibers can be deposited either as a nonwoven mat or as a highly aligned bundle of controllable size. Potential use of this nanofibrous matrix for tissue engineering was studied by examining its integrity in water and cellular compatibility. It was found that the matrix with a chitosan/PEO ratio of 90/10 retained excellent integrity of the fibrous structure in water. Experimental results from cell stain assay and SEM imaging showed that the nanofibrous structure promoted the attachment of human osteoblasts and chondrocytes and maintained characteristic cell morphology and viability throughout the period of study. This nanofibrous matrix is of particular interest in tissue engineering for controlled drug release and tissue remodeling. PMID- 15885771 TI - Extent of iron pick-up in deforoxamine-coupled polyurethane materials for therapy of chronic wounds. AB - Polyurethane net substrates (PNS) coupled with deferoxamine (DFO) have been studied to determine the extent of Fe2+ pick-up for use in chronic wound therapy. A m solution of ferrous sulphate (FeSO4) was used to generate ferrous ions similar to those found in chronic wounds. The concentration of Fe as a function of position through the dressings was evaluated using a variety of techniques. Atomic force microscopy (AFM) and energy-filtered transmission electron microscopy (EFTEM) revealed a rough precipitated layer at the surface of activated PNS exposed to FeSO4 solution. Optical microscopy (OM) and backscattered environmental scanning electron microscopy (ESEM) showed a clear layer of Fe(3+)-enriched material in the surface regions exposed to DFO. The penetration depth of DFO into activated dressings was found to be 20-30 microm. Energy-dispersive X-ray (EDX) analysis was used to approximate the distribution of bound- and unbound-Fe as a function of position within BPNS and DFO-activated dressings after immersing them in a FeSO4 solution for various times. These studies have shown the activity of iron with respect to ionic state in DFO activated PNS for potential using as dressing for chronic wounds. PMID- 15885772 TI - Synthesis and characterization of porous beta-tricalcium phosphate blocks. AB - Porous beta-tricalcium phosphate (beta-TCP) blocks with four different macropore sizes (pore larger than 50 microm were synthesized using "calcium phosphate emulsions", and characterized by optical, geometrical, gravimetric, and radiological methods. The reproducibility of the synthesis method was excellent. Moreover, the macropore size could be easily controlled without modifying the microporosity (pore smaller than 50 microm) or the total porosity (microporosity+macroporosity). Based on the initial composition of the blocks and their final apparent density, the microporosity, macroporosity, and the total block porosity were calculated to be close to 21%, 54%, and 75%, respectively. These values were confirmed by microcomputed tomography (microCT). The mean macropore diameters were close to 150, 260, 510 and 1220 microm, as measured optically. Consistently lower values (25% lower) were obtained by microCT, but the linear correlation between microCT and optical method was high (r(2)>0.97). The macropore size distribution calculated from microCT scans appears to be narrow and normally distributed. The very good correlation between the results of the various methods and the possibility to determine the pore size distribution suggest that microCT is an ideal tool to non-destructively characterize macroporous calcium phosphate bone substitutes. PMID- 15885773 TI - Galanin and alcohol dependence: neurobehavioral research. AB - It is known that microinjection of galanin (GAL) intraventricularly or in specific hypothalamic sites increases food consumption and, conversely, the intake of food increases the expression of GAL in hypothalamic sites. Ethanol (EtOH) is a calorie-rich food as well as a drug of abuse. The research reviewed here shows that GAL may play a similar role in alcohol intake. First, experiments in which GAL was microinjected into the third ventricle or the paraventricular nucleus (PVN) showed increases in EtOH consumption. The increase in EtOH consumption occurred during both the light and dark cycles after GAL injection in the third ventricle in rats with limited EtOH access. Injection of GAL did not increase food intake in rats that had been chronically drinking alcohol. GAL receptor blockade reversed these increases. Microinjection of GAL directly into the PVN also increased ad libitum EtOH intake and blockade of these receptors in the PVN inhibited ad libitum EtOH consumption. Secondly, rats administered EtOH showed increases in GAL in the PVN and related hypothalamic sites. EtOH injection and voluntary intake, both ad libitum and limited access, increased GAL gene and peptide expression in the PVN consistently across administration procedures. These experiments show that GAL injection increases alcohol intake and that the intake of alcohol increases GAL, suggesting a positive feedback relationship between alcohol intake and specific hypothalamic GAL systems. Such a relationship may contribute to the motivation to consume excessive alcoholic beverages and the development of alcohol dependence. PMID- 15885774 TI - Constitutive and ligand-induced internalization of EGFP-tagged galanin R2 and Rl receptors in PC12 cells. AB - In the present experiments trafficking of the galanin R1-(GALR1) and, in particular, the galanin R2 receptor (GALR2) was studied after fusion with enhanced green fluorescent protein (GALR1-EGFP and GALR2-EGFP) and transfection into PC12 cells. Both fusion proteins were predominantly localized on the plasma membrane and internalized in a dose dependent manner after incubation with galanin. Preincubation with M35 or M40 did not prevent galanin-induced internalization of GALR1-EGFP or GALR2-EGFP. However, AR-M1896, a selective GALR2 agonist, caused GALR2, but not GALR1 internalization. Hyperosmotic sucrose inhibited internalization of GALR2-EGFP. After co-incubation with galanin, GALR2 EGFP was co-localized with internalized Texas Red transferrin, a marker of the clathrin endocytic pathway. Experiments with protein synthesis inhibition and Texas Red transferrin suggest that GALR2 is constitutively internalized. Studies in progress will show if this is the case also for GALR1. PMID- 15885775 TI - Expression of AgRP, NPY, POMC and CART in human fetal and adult hippocampus. AB - The Agouti-Related Protein (AgRP), Neuropeptide Y (NPY), Proopiomelanocortin (POMC) and the Cocaine and Amphetamine-Regulated Transcript (CART) are four neuropeptides that play essential roles in the regulation of food intake and energy homeostasis in mammals. CART, POMC and NPY have also been suggested to play a role in the development of the hippocampus. We therefore employed quantitative real-time RT-PCR (qPCR) to analyze the expression levels of these genes in the fetal and adult human hippocampus to examine whether the four neuropeptides are differentially regulated in the hippocampus during development. CART (6.5-fold) and POMC (8.3-fold) mRNAs were significantly higher in the adult hippocampus. NPY on the other hand, was significantly reduced (2.1-fold) in the adult hippocampus, while AgRP mRNA was comparatively unchanged between fetal and adult hippocampus. In relative terms, CART mRNA was the highest and AgRP the lowest in both the fetal and adult hippocampus. CART, POMC and NPY are, therefore, differentially expressed in the human fetal and adult hippocampus and could play a role in its development or could be regulated by various stimuli involved in the development of this brain structure. PMID- 15885776 TI - CD 39-associated high ATPase activity contribute to the loss of P 2 X 7-mediated calcium response in LCL cells. AB - The P 2 X 7 nucleotide receptor is an adenosine 5'-triphosphate (ATP)-gated ion channel, which induces cation channel opening imparting significant permeability to Ca(2+), and is widely expressed in cells of hematopoietic origin. Our previous report showed that P 2 X 7-mediated calcium response was absent in three Epstein Barr virus (EBV)-positive and P 2 X 7 positive cell lines. In this report, we detected the cell surface ATPase activity, which contributes to the hydrolysis of extracellular ATP, and the expression of CD 39, which is the main source of ATPase on hematopoietic cells, in these cell lines. Then, we tried to restore the P 2 X 7-mediated calcium response in LCL-H and J 6-1 cells by either increasing the concentration of agonist or suppressing the ATPase activity by betagammaMeATP, a synthetic poorly metabolizable ATP analogue. The results showed that LCL-H and J 6-1 cells had higher levels of ATPase activity and CD 39 expression. The treatment of 300 microM betagammaMeATP efficiently inhibited the ATPase activity on LCL-H and J 6-1 cells. Both elevation of agonist concentration (10mM ATP or 1mM BzATP) and pretreatment with 300 microM betagammaMeATP followed by stimulation with normal concentration of agonists (1mM ATP or 0.1mM BzATP) could cause P 2 X 7-mediated calcium response in LCL-H but neither in J 6-1 cells. These results suggested that multiple mechanisms contributed to the loss of the P 2 X 7-mediated calcium response. CD 39-associated high ATPase activity contributed to the loss of the P 2 X 7-mediated calcium response in LCL-H cells, while additional mechanism(s) existed in J 6-1 cells. PMID- 15885777 TI - Cytokines as mediators of depression: what can we learn from animal studies? AB - It has recently been postulated that cytokines may cause depressive illness in man. This hypothesis is based on the following observations: 1. Treatment of patients with cytokines can produce symptoms of depression; 2. Activation of the immune system is observed in many depressed patients; 3. Depression occurs more frequently in those with medical disorders associated with immune dysfunction; 4. Activation of the immune system, and administration of endotoxin (LPS) or interleukin-1 (IL-1) to animals induces sickness behavior, which resembles depression, and chronic treatment with antidepressants has been shown to inhibit sickness behavior induced by LPS; 5. Several cytokines can activate the hypothalamo-pituitary-adrenocortical axis (HPAA), which is commonly activated in depressed patients; 6. Some cytokines activates cerebral noradrenergic systems, also commonly observed in depressed patients; 7. Some cytokines activate brain serotonergic systems, which have been implicated in major depressive illness and its treatment. The evidence for each of these tenets is reviewed and evaluated along with the effects of cytokines in classical animal tests of depression. Although certain sickness behaviors resemble the symptoms of depression, they are not identical and each has distinct features. Thus the value of sickness behavior as an animal model of major depressive disorder is limited, so that care should be taken in extrapolating results from the model to the human disorder. Nevertheless, the model may provide insight into the etiology and the mechanisms underlying some symptoms of major depressive disorder. It is concluded that immune activation and cytokines may be involved in depressive symptoms in some patients. However, cytokines do not appear to be essential mediators of depressive illness. PMID- 15885778 TI - Maximum tolerated dose of a humanized anti-vascular endothelial growth factor antibody fragment for treating neovascular age-related macular degeneration. AB - PURPOSE: To investigate the maximum tolerated dose of ranibizumab administered as a single intravitreal injection. DESIGN: Open-label, 5-center, uncontrolled, prospective, dose-ranging, interventional case series. PARTICIPANTS: Twenty-seven patients with subfoveal choroidal neovascularization (CNV) secondary to age related macular degeneration (AMD) with best-corrected Snellen equivalent visual acuity (VA) of 20/100 or worse and considered ineligible for laser photocoagulation or photodynamic therapy. METHODS: A single intravitreal injection of ranibizumab was to be administered at 1 of 6 escalating doses (50, 150, 300, 500, 1000, and 2000 microg), with escalation to the next dose level occurring only after the safety and tolerability of the lower dose level was established through postinjection day 14. Follow-up examinations were performed on postinjection days 1, 3, 7, 14, 42, and 90. Enrollment was stopped if > or =2 patients experienced dose-limiting toxicity. MAIN OUTCOME MEASURES: The primary safety measures were changes from baseline in VA, intraocular pressure (IOP), intraocular inflammation, and production of antiranibizumab antibody. Dose limiting toxicity was defined by intraocular inflammation, elevated IOP, reduced VA, or hemorrhage within 90 days after injection. RESULTS: All patients completed this single intravitreal injection study, and 500 microg of ranibizumab was the maximum tolerated dose. At the higher dose of 1000 microg, significant intraocular inflammation was noted. All adverse events were self-limited, and no infectious endophthalmitis occurred. Aqueous or vitreous ocular inflammation occurred in 12 subjects, with complete resolution within 42 days. In 9 of the subjects, the inflammation was graded as trace to 1+ and required no treatment; in 3 of the subjects, the inflammation was graded as 2+ or 3+, and 2 of the 3 were treated with topical 1% prednisolone acetate. No serum antiranibizumab antibodies were detected. All patients had VA similar or improved compared with baseline values. CONCLUSION: The maximum tolerated single dose of ranibizumab in neovascular AMD patients was 500 microg. Single intravitreal injections of ranibizumab up to a dose of 500 microg were safe and well tolerated in this small group of patients. PMID- 15885779 TI - Cavitary changes in retinoblastoma: relationship to chemoresistance. AB - OBJECTIVE: To describe the clinical features and outcome of treatment of retinoblastomas presenting with ophthalmoscopically visible cavities. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Sixteen eyes of 15 patients harboring retinoblastomas with ophthalmoscopically visible cavities treated between February 1995 and July 2003. METHODS: Patient demographics, eye and tumor characteristics, and response to treatment were recorded. MAIN OUTCOME MEASURES: Decrease in tumor size and tumor recurrence. RESULTS: The mean patient age at diagnosis was 20 months. The mean number of retinoblastomas per eye was 2 (range, 1-6). Associated subretinal fluid was graded as absent in 5 eyes, minimal in 6 eyes, mild in 4 eyes, and severe in 1 eye. None of the tumors had vitreous seeds, and 5 (31%) had subretinal seeds. Ophthalmoscopically visible calcification occupied a mean of only 3% of the tumor (median, 0%; range, 0%-30%). Intratumoral cavities were characteristically visible in the superficial portion of the tumor and had a mean diameter of 2.0 mm (median, 1.5; range, 0.5-5.0). The mean number of cavities per tumor was 2 (median, 1; range, 1-5). Three tumors were treated with enucleation, 1 with observation, and 12 with chemoreduction. Two months after chemoreduction, the mean reduction in tumor base was 19% (median, 16%; range, 0%-42%), and the mean reduction in tumor thickness was 18% (median, 12%; range, 0%-59%). During a mean follow-up of 32 months (median, 23; range, 12-58), none of the cavitary retinoblastomas recurred or gave rise to vitreous or new subretinal seeds. CONCLUSION: Retinoblastomas containing ophthalmoscopically visible cavitary spaces do not show a substantial decrease in size after chemotherapy. Cavitary changes in retinoblastoma may be a sign that the tumor is well differentiated and may impart a better prognosis. PMID- 15885780 TI - Poor prognosis of visual outcome after visual loss from giant cell arteritis. AB - OBJECTIVE: To evaluate the incidence and extent of visual recovery in patients with giant cell arteritis (GCA) treated with high doses of systemic corticosteroids. DESIGN: Multicenter prospective case series. PARTICIPANTS: Thirty-four consecutive patients with biopsy-proven GCA and visual loss seen at either Wills Eye Hospital in Philadelphia or Auckland Hospital in New Zealand from 2001 to 2004 were evaluated prospectively. METHODS: All patients underwent a complete neuro-ophthalmic evaluation that included Snellen visual acuity, pseudoisochromatic Ishihara color plates, visual fields, intraocular pressure, slit-lamp examination, and binocular ophthalmoscopy. Formal visual field testing was performed on all patients who were capable of completing the test. All patients were treated with a standard protocol of 1 g of IV methylprednisolone daily for 3 days followed by oral prednisone 60 or 80 mg (depending on patient weight). Patients were evaluated initially at 2-week intervals (14-19 days) and then monthly (4-5 weeks), with subsequent dosages of prednisone modified based on erythrocyte sedimentation rate, C-reactive protein, and patient symptoms. At each visit, patients underwent a repeat complete neuro-ophthalmic evaluation. MAIN OUTCOME MEASURES: Alterations in visual acuity, visual field, and color vision. RESULTS: Patients with visual loss from GCA had a mean visual acuity of 20/400. Visual deterioration occurred in 27% of eyes within the first week despite high dose IV corticosteroids. Fifteen percent of eyes showed an improvement of visual acuity within the first month, but only 5% have corresponding improvement in visual field. CONCLUSIONS: Visual recovery is uncommon in patients who lose vision from GCA. Recovery in visual acuity is not associated with visual field or color vision improvement in this series. Visual deterioration occurs in approximately 27% of eyes despite high-dose IV methylprednisolone. The greatest risk of visual deterioration is in the first 6 days. PMID- 15885781 TI - Ptosis and orbital fat prolapse after posterior sub-Tenon's capsule triamcinolone injection. AB - PURPOSE: To describe the occurrence of orbital fat prolapse and blepharoptosis after posterior sub-Tenon (PST) triamcinolone injection. DESIGN: Retrospective review of consecutive case series. PARTICIPANTS: Patients with ptosis and orbital fat herniation after PST triamcinolone injection. METHODS: Charts of all patients with ptosis and orbital fat herniation presenting after PST triamcinolone injection to the oculoplastics service of the Cole Eye Institute between 1999 and 2003 were reviewed. Charts were reviewed for patient age, indication, dates of injections, time to patient complaint or time to referral for ptosis, and marginal reflex distance (MRD1). MAIN OUTCOME MEASURES: Ptosis and orbital fat herniation after PST triamcinolone injection. RESULTS: Eleven patients with a history of ipsilateral PST triamcinolone injections were seen with ptosis and orbital fat prolapse. Ten charts were available for review. Mean patient age was 64 years (range, 45-78 years). Patients underwent 1 to 9 ipsilateral injections, and 2 patients underwent bilateral injections. Patients were seen for ptosis evaluation on average 22.5 months (range, 3-56 months) after the initial injection, and 6.6 months (range, 0-20 months) after the most recent injection. All patients demonstrated significant orbital fat prolapse in conjunction with statistically significant ptosis (P = 0.016). Tissue was obtained in 3 cases. Histologic findings in 1 case showed orbital fat infiltrated by histiocytes that seemed to contain phagocytosed material. CONCLUSIONS: Posterior sub-Tenon triamcinolone injection may cause ptosis associated with orbital fat prolapse. This finding may be a relatively common complication of PST triamcinolone injection. We recommend counseling patients about this risk before PST triamcinolone injection. PMID- 15885782 TI - Microkeratome-assisted lamellar keratoplasty for the surgical treatment of keratoconus. AB - PURPOSE: To evaluate the visual and refractive results of microkeratome-assisted lamellar keratoplasty (LK) performed on keratoconus patients intolerant to spectacles and contact lenses. DESIGN: Prospective, noncomparative, interventional study. PARTICIPANTS: A microkeratome-assisted LK procedure was performed on 50 eyes of 50 keratoconus patients. All patients were spectacle and contact lens intolerant. INTERVENTION: All patients included in this study underwent a standard surgical procedure involving removal of a lamella (9 mm in diameter cut with the 250-microm microkeratome head) from the recipient cornea by means of a hand-driven microkeratome and suturing of a donor lamella (0.5 mm smaller in diameter than the removed corneal lamella, cut with the 350-microm microkeratome head) obtained from a cornea mounted on an artificial anterior chamber. Each patient was examined preoperatively and at different postoperative times (1 and 6 months and 1, 2, 3, and 4 years). MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), 1-year best contact lens-corrected visual acuity (BCLCVA), refraction, and computerized analysis of corneal topography. RESULTS: After suture removal was completed, both UCVA and best-corrected visual acuity were significantly improved over properative values at all examination times. One year postoperatively, when follow-up was still available for all patients, UCVA was better than 20/200 in 8 of 50 (16%) patients and BSCVA was > or =20/40 in 44 of 50 (88%) patients, whereas BCLCVA was > or =20/40 in all 50 patients. Refractive astigmatism within 4 diopters was seen in 43 of 50 (86%) patients. Corneal topographic patterns were classified as regularly astigmatic in 39 of 50 (78%) patients. The 1-year values did not change substantially at later postoperative examination times. Complications included preparation of donor grafts of poor quality that needed to be discarded (8 cases [16%]), irregular astigmatism of various degrees (11 cases [22%]), high-degree astigmatism requiring secondary intervention (6 cases [12%]), epithelial interface ingrowth (1 case [2%]), and cataract formation (1 case [2%]). CONCLUSIONS: Microkeratome-assisted LK can be performed on corneas with moderate to advanced keratoconus with a minimal corneal thickness of >380 microm. The procedure is relatively simple, may be standardized in most of its parts, and does not involve time-consuming maneuvers. All complications recorded did not threaten vision and were dealt with successfully. Our results indicate that microkeratome-assisted LK is as efficacious as conventional penetrating keratoplasty for the surgical treatment of keratoconus. However, the time necessary to achieve stable results is considerably shorter. PMID- 15885783 TI - Ocular complications of hypovitaminosis a after bariatric surgery. AB - PURPOSE: To report the ocular complications of xerophthalmia, nyctalopia, and visual deterioration to legal blindness as a result of inadequate vitamin A supplementation after malabsorptive bariatric surgery. DESIGN: Observational case report and literature review. PARTICIPANT: A 39-year-old woman with xerophthalmia and nyctalopia occurring 3 years after gastric bypass surgery. RESULTS: We report a patient with a rare finding of xerophthalmia and visual deterioration after gastric bypass surgery as a result of vitamin A deficiency. The patient was referred for decreased vision associated with chronic dry eyes, bilateral diffuse punctate keratitis, and corneal scarring of unknown cause after several ophthalmologic examinations. The medical history, ophthalmic findings, and clinical course are discussed. CONCLUSIONS: Gastric bypass procedures can cause vitamin A deficiency leading to serious ocular complications, including xerophthalmia, nyctalopia, and ultimate blindness. The increasing incidence of obesity and gastric bypass procedures warrants patient and physician education regarding strict adherence to vitamin supplementation. Education is imperative to avoid detrimental ophthalmic complications resulting from hypovitaminosis A and to prevent a potential epidemic of iatrogenic xerophthalmia and blindness. PMID- 15885784 TI - Cataract surgery for residual angle closure after peripheral laser iridotomy. AB - PURPOSE: To investigate the frequency of residual angle closure after resolution of pupillary blocking by laser peripheral iridotomy and the effects of subsequent cataract surgery to resolve angle closure completely. DESIGN: Retrospective, consecutive, interventional study. PARTICIPANTS: Among 70 eyes treated with laser iridotomy, 13 with residual angle closure were treated with cataract surgery. METHODS: The provocative test of angle closure by prone position in a dark room for 1 hour was performed; increases in tension of > or =8 mmHg, 6 or 7 mmHg, and < or =5 mmHg were considered to be positive, suspected positive, and negative, respectively. Configuration of the anterior chamber was examined using ultrasound biomicroscopy (UBM). MAIN OUTCOME MEASURES: Intraocular pressure (IOP), response to the dark room prone position test, and morphologic analysis by UBM were evaluated before and 3 months after cataract surgery. RESULTS: Residual angle closure after iridotomy was seen in 27 (38.6%) of 70 eyes; this was confirmed functionally by the dark room prone position test and morphologically by UBM. Eyes with IOP of > or =20 mmHg or with a glaucomatous visual field defect before iridotomy had a significantly higher incidence of residual angle closure after iridotomy than eyes without these findings (P<0.05). In all the eyes with residual angle closure after iridotomy, the response to the prone position test became negative after cataract surgery, with significant lowering of IOP (P<0.01). CONCLUSIONS: Residual angle closure after iridotomy was common, especially in eyes with primary angle closure and poorly controlled IOP or glaucomatous optic neuropathy. Cataract surgery was effective to resolve completely the residual angle closure after iridotomy and lower IOP. PMID- 15885785 TI - A unique corneal dystrophy of Bowman's layer and stroma associated with the Gly623Asp mutation in the transforming growth factor beta-induced (TGFBI) gene. AB - PURPOSE: To report a unique corneal dystrophy characterized by deposits at Bowman's layer and stromal lattice lines associated with the Gly623Asp missense mutation in the transforming growth factor beta-induced (TGFBI) gene. DESIGN: Experimental study. PARTICIPANTS AND CONTROLS: The proband, 3 affected siblings, 4 unaffected relatives, and 100 control individuals. METHODS: Slit-lamp examination, photographic documentation, and isolation of genomic DNA from buccal mucosal swabs obtained from each family member examined. Exons 4 and 11 to 14 of the TGFBI gene were amplified and sequenced in these family members and in control individuals. MAIN OUTCOME MEASURES: Clinical characteristics of corneal opacification in affected patients and presence of coding region changes in the TGFBI gene. RESULTS: Significant phenotypic variability, including polymorphic Bowman's layer opacities and stromal lattice lines, was noted in the 4 affected siblings who were examined. Screening of TGFBI exon 14 in the proband, 3 affected siblings, and a 19-year-old unaffected relative revealed a missense change, Gly623Asp, that was absent in the other 3 unaffected relatives screened and in 200 control chromosomes. CONCLUSIONS: We report a novel corneal dystrophy phenotype secondary to the Gly623Asp mutation in the TGFBI gene that is associated with clinical features of both lattice corneal dystrophy and a Bowman's layer dystrophy. The presence of clinical features considered atypical for a TGFBI-associated dystrophy in this pedigree, as well as the wide range of phenotypic expressions of the Gly623Asp mutation in affected members, underscore the clinical utility of molecular genetic analysis in the diagnosis of suspected corneal dystrophies. PMID- 15885786 TI - Comparison of arteritis and nonarteritic anterior ischemic optic neuropathies with the Heidelberg Retina Tomograph. AB - OBJECTIVE: Quantitative data are not available regarding the morphological appearance of optic nerve morphology after arteritic anterior ischemic optic neuropathy (AAION) and nonarteritic anterior ischemic optic neuropathy (NAION). The purpose of this study is to evaluate the appearance of the optic nerve head in patients after AAION and NAION with the Heidelberg Retina Tomograph (HRT). DESIGN: Prospective comparative observational study. PARTICIPANTS: Thirty-eight patients with either unilateral AAION (15) or NAION (23). METHODS: Heidelberg Retina Tomograph images were taken of both affected and unaffected eyes. The quantitative changes in the optic nerve head were morphologically evaluated and compared with those of the normal contralateral eyes. MAIN OUTCOME MEASURES: Heidelberg Retina Tomograph parameters were analyzed using the uninvolved contralateral eye as a control. RESULTS: The ratio between cup area and disc area was 42% larger in the affected eyes in the NAION group than in the fellow eyes (P = 0.03, t test). In the AAION patients, the ratio between cup area and disc area was 116% larger in involved eyes than in their fellow eyes (P = 0.0003, t test). In addition, in the AAION group, relative to their fellow eyes, the rim area was 27% smaller (P = 0.0006), cup volume was 160% larger (P = 0.021, t test), mean cup depth was 29% larger, cup shape measure differed by 41% (suggesting more excavation), and mean retinal nerve fiber layer thickness was 30% less. Relative to the fellow eyes in the NAION group, the rim area was 6% smaller (P = 0.13, Student's t test), and the cup shape measure was 13% different, although these were not statistically significant. When the uninvolved eyes of the NAION and AAION groups were compared directly, there was no statistical difference between any of the HRT parameters, except mean cup depth (0.09 vs. 0.14 mm, respectively; P = 0.02). CONCLUSIONS: This study is the first to demonstrate quantitatively an enlargement and excavation of the optic cup after an event of AAION, but not NAION. The eyes affected with AAION showed significant excavation and enlargement of the optic cup when compared with contralateral uninvolved eyes. All HRT generated parameters showed changes consistent with increased enlargement and excavation of AAION-involved eyes. These changes were not observed in eyes that experienced an event of NAION. PMID- 15885787 TI - Differential expression of vascular endothelial growth factor implies the limbal origin of pterygia. AB - OBJECTIVE: To study the distribution of isoforms of vascular endothelial growth factor (VEGF) and its receptors VEGFR1 and VEGFR2 in pterygia and to compare it with that in healthy conjunctivas. DESIGN: Nonrandomized comparative (cadaver controlled) study with histopathologic correlations. METHODS: Tissue specimens from 75 patients treated for primary pterygia were analyzed using immunohistochemical studies as well as different molecular biological examinations. Healthy conjunctivas from 33 patients treated for cataracts as well as specimens from the conjunctiva, limbus, and lens of both eyes of 12 body donors served as controls. TESTING: Surgical specimens of pterygia and normal conjunctiva specimens were processed with paraffin, sectioned, stained using specific antibodies against VEGF and its receptors, and examined by light microscopy. The other part of both groups of specimens as well as specimens from body donors were prepared and analyzed by means of reverse-transcription polymerase chain reaction (RT-PCR), real-time RT-PCR, enzyme-linked immunosorbent assay, and Western blots. MAIN OUTCOME PARAMETERS: Vascular endothelial growth factor and VEGFR1 and VEGFR2 were analyzed to indentify the splice variants of VEGF as well as the distribution and amount of VEGF and both receptors in pterygia and the control tissues. RESULTS: In analysis of specimens from pterygium patients as well as normal conjunctivas, VEGF121 and VEGF165 were identified as the only VEGF splice forms expressed. In addition to VEGF, VEGFR1 and VEGFR2 were detected in pterygia and conjunctivas and immunostained within the epithelium of pterygia and conjunctivas and on intrapterygial and intraconjunctival endothelial cells. Levels of VEGFR1 and VEGFR2 mRNA were lower in pterygia than in conjunctivas but similar in limbal and pterygium samples. Vascular endothelial growth factor levels were higher in pterygia than in conjunctivas, but were similar in the limbus and pterygia. CONCLUSIONS: The results reveal similar behaviors in limbal and pterygium epithelial cells in terms of VEGF and VEGFR expression, with the presumption that pterygia arise from limbal epithelial cells and that human conjunctivas are not a suitable control for the analysis of pterygia. Moreover, the results suggest that VEGF might play an active role in the physiology of conjunctival epithelial cells. PMID- 15885788 TI - The effect of periocular warming on accommodation. AB - PURPOSE: To investigate the effect of periocular warming on accommodation amplitude and near vision after prolonged near work in visual display terminal (VDT) workers in an office setting. DESIGN: Prospective interventional observational case series. PARTICIPANTS: Ten healthy Japanese male volunteers (mean age, 39.3+/-2 years [range, 36-43]) with a best distance corrected visual acuity (VA) of >20/20 who performed near work for 7 hours were included in the study. MAIN OUTCOME MEASURES: The differences of subjective and objective accommodations, near acuity, and pupil diameters were compared and studied in 2 experimental settings with and without periocular warming. METHODS: Two experiments were performed on each subject on 2 different days. In one experiment, the subjects wore an eye mask and closed their eyelids for 10 minutes after 7 hours of VDT work (lid closure [LC] group). In the other experiment, the subjects applied warming sheets (WSs) to their eyelids and the periocular region and wore masks for 10 minutes again after 7 hours of VDT work (WS group). Visual display terminal work was performed under the same settings on both days. Subjective and objective accommodations, near vision, and pupil diameters were measured immediately before and immediately after the resting or warming procedure and again 90 minutes after the resting or warming procedures. RESULTS: Fifty percent of eyes had an increase in the accommodation amplitude of at least 0.5 diopters immediately after application of the WSs. Forty percent of eyes still maintained the accommodation amplitude level at 90 minutes. The mean near logarithm of the minimum angle of resolution VA immediately after application of WSs and at 90 minutes significantly improved relative to the near acuity values after simple eyelid closure. CONCLUSION: Periocular warming was found to be associated with increases in subjective and objective accommodations, with a concomitant improvement in near VA. Periocular warming with disposable WSs is a simple and effective method of increasing near work-related ergonomy. PMID- 15885789 TI - Posterior sub-Tenon's capsule triamcinolone injection combined with focal laser photocoagulation for diabetic macular edema. AB - PURPOSE: To investigate the efficacy of posterior sub-Tenon's capsule triamcinolone injection (PSTI) combined with focal laser (FL) photocoagulation in treatment of diffuse clinically significant diabetic macular edema (CSME). DESIGN: Prospective randomized clinical study. PARTICIPANTS: Sixty patients with diffuse CSME. METHODS: We randomized the patients who had CSME into 2 treatment groups. A total of 30 eyes received macular focal and grid laser photocoagulation (MP), and the second group of 30 eyes received FL photocoagulation combined with PSTI (FL + PSTI). Clinical and visual evaluations were performed at baseline and 12 and 18 weeks after treatment, and the results were compared by statistical methods. MEAN OUTCOME MEASURES: Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity (VA) scores and clinical changes in macular edema. RESULTS: Before treatment, mean ETDRS VA scores were 45.9+/-12.7 (mean +/- standard deviation [SD]; range, 24-66) in MP (focal + grid laser) and 44.5+/-12.6 (range, 22-66) in the FL + PSTI group. Early Treatment Diabetic Retinopathy Study scores at 18 weeks were 46.4+/-14.2 (range, 20-70) after MP and 52.5+/-17.5 (range, 22 82) after FL + PSTI. Early Treatment Diabetic Retinopathy Study scores significantly improved in eyes that received FL + PSTI (P = 0.0001, paired t test). The FL + PSTI group had an increase of 12.4+/-7.1 (range, 5-26) ETDRS letters, and the MP group had an increase of 7.8+/-3.1 (range, 5-15) letters (P = 0.04, unpaired t test). Twelve of 30 (40%) cases in the MP group and 24 of 30 (80%) cases in the FL + PSTI group showed clinical improvement in macular edema according to the stereoscopic evaluation of fundus and fluorescein angiography (P = 0.003). CONCLUSION: Posterior sub-Tenon's capsule application of triamcinolone may improve early visual outcome in diffuse diabetic macular edema when combined with FL photocoagulation. PMID- 15885790 TI - Surgical anatomy of the lacrimal fossa a prospective computed tomodensitometry scan analysis. AB - PURPOSE: To establish the accurate surgical anatomy of endonasal dacryocystorhinostomy (DCR) based on the radiological analysis of underlying bony structures. DESIGN: Prospective noncomparative observational case series study. PARTICIPANTS: Fifty-nine patients with complete nasolacrimal stenosis underwent a computed tomodensitometry (CT) scan before endonasal DCR. METHODS: High resolution CT scanning with contrast injection of the lacrimal sac was performed. Image reconstruction was performed to obtain continuous 1.0-mm axial and coronal sections for review. MAIN OUTCOME MEASURES: Relationship of the lacrimal fossa (LF) to the operculum of the middle turbinate (OMT), the uncinate process (UP), and the frontal recess (FR); symmetry of the right and left anatomies; location of the OMT; position of the most anterior insertion of the UP with respect to 2 main references (the posterior lacrimal crest and the junction between the maxillary and lacrimal bones) on axial sections at 3 different levels (upper, intermediate, and lower of the LF); height of the LF; and distance of the OMT from the lower limit of the LF. RESULTS: The OMT, the UP, and the FR were adjacent to the LF in 41 (53.2%), 73 (94.8%), and 23 cases (29.9%), respectively. There was a right-left symmetry in 10 of 18 patients (55%). The OMT was always anterior to the junction between the maxillary bone and the lacrimal bone. The UP was more frequently posterior (32.5%) or adjacent (45.5%) to the LF at the lower level, adjacent to the maxillary bone (55.8%) at the intermediate level, and adjacent to the middle turbinate (61%) at the upper level. The height of the LF was 12.06+/-1.93 mm. The OMT was located 5.96+/-2.05 mm upward from the lower limit of the LF. CONCLUSION: The almost constant overlapping of the UP onto the LF at the level of the common canaliculus indicates that the most effective approach for successful DCR osteotomy is via a submucosal cleavage and resection of the anterior part of the UP. The management of these landmark structures should be an integral part of the endonasal DCR method. PMID- 15885791 TI - Subconjunctival carboplatin in fibrin sealant in the treatment of transgenic murine retinoblastoma. AB - PURPOSE: To evaluate the efficacy of subconjunctival carboplatin in fibrin sealant in the treatment of transgenic murine retinoblastoma. DESIGN: Experimental study using LHbeta-Tag transgenic mice in a randomized controlled trial. PARTICIPANTS AND CONTROLS: Thirty-three 10-week-old LHbeta-Tag transgenic mice: 22 carboplatin-treated animals and 11 control animals. METHODS: Three groups of 11 mice were treated with a single, 30 microl injection of fibrin sealant in the subconjunctival space of 1 eye; the opposite eye was left untreated as an internal control. Group 1 (low-dose group) received 37.5 mg/ml calculated concentration of carboplatin in fibrin sealant (0.66 mg measured total dose). Group 2 (high-dose group) received 75 mg/ml calculated concentration of carboplatin in fibrin sealant (1.23 mg measured total dose). Group 3 (control group) received fibrin sealant only. Mice were killed on day 22 after treatment. Eyes were serially sectioned, and retinal tumor burden was quantified by histopathologic analysis. For statistical analysis of treatment effects, eyes were divided into 6 groups: low-dose group, sealant-treated eyes; low-dose group, untreated eyes; high-dose group, sealant-treated eyes; high-dose group, untreated eyes; control group, sealant-treated eyes; and control group, untreated eyes. MAIN OUTCOME MEASURES: Main outcome measure was mean tumor burden per level per eye in each experimental group. RESULTS: The best therapeutic results were obtained in eyes treated with low-dose carboplatin in fibrin sealant, where no histopathologic evidence of toxicity was observed, and 6 of 11 eyes had zero tumor burden. Tumor burden in the remaining 5 eyes in this group was minimal (4 eyes) or moderate (1 eye) compared with mean control values. Mean tumor burden in this group was significantly smaller than mean tumor burden in untreated eyes from the same mice (P<0.004), sealant-treated eyes in the control group (P<0.004), and untreated eyes in the control group (P<0.002). Although a similar reduction in mean tumor burden was observed in eyes treated with high-dose carboplatin in fibrin sealant, 5 of 10 eyes analyzed in this group also demonstrated histopathologic evidence of severe toxicity. CONCLUSIONS: Subconjunctival carboplatin in fibrin sealant is effective in the treatment of transgenic murine retinoblastoma. A single injection of low-dose carboplatin in fibrin sealant was sufficient to induce complete or near-complete intraocular tumor regression in 10 of 11 eyes (91%), with no associated histologic evidence of toxicity. These results suggest that subconjunctival carboplatin in fibrin sealant provides sustained release and could have clinical use in the treatment of intraocular retinoblastoma. PMID- 15885792 TI - Tumor, node, metastasis classification of malignant ciliary body and choroidal melanoma evaluation of the 6th edition and future directions. AB - PURPOSE: The tumor, node, metastasis classification of malignant uveal melanoma has been revised. We evaluated how the 6th edition (TNM6) improves on the previous one (TNM5). DESIGN: Population-based, retrospective, cross-sectional study. PARTICIPANTS: Two hundred eighty-nine consecutive patients who had a ciliary body and choroidal melanoma treated in the district of the Helsinki University Central Hospital, Finland, between 1962 and 1981. METHODS: Tumor dimensions, ciliary body involvement, and extraocular extension were evaluated from histopathologic sections and pathology reports. Tumors were assigned into categories and stages according to TNM6, TNM5, and 2 previously proposed size classifications. MAIN OUTCOME MEASURES: Proportion of tumors classified in each category and melanoma-specific survival by category and stage. RESULTS: Of the 289 melanomas, 5% were classified as pT1, 63% as pT2, 22% as pT3, and 7% as pT4 according to TNM6. The corresponding percentages based on TNM5 were 8%, 17%, 63%, and 10%. Of pT2 tumors in TNM6, 4% came from pT1, 65% from pT3, and 4% from pT4 category of TNM5. Of 28 melanomas with extraocular growth, 29% were classified as pT2 in TNM6 rather than pT4 in TNM5. The 10-year survival estimate was 2 percentage points lower for pT1, 7 percentage points higher for pT2, 17 percentage points lower for pT3, and 13 percentage points lower for pT4 by TNM6 compared with TNM5; TNM6 (P<0.0001) and the modified alternative size classifications (P = 0.0022 and P = 0.0026) divided tumors according to prognosis better than TNM5. The 10-year survival for stage I, II, and III tumors was 2 percentage points lower, 7 points higher, and 23 points lower by TNM6, which was not better than TNM5 in separating patients according to prognosis (P = 0.47). The alternative size classifications provided more equal categories and fitted the data set better than TNM5 regarding prognosis. CONCLUSIONS: TNM6 is an improvement over TNM5 in some, but not all, respects. Areas for development include taking into account ciliary body involvement and extraocular extension in more detail and combining into each stage tumor categories with similar rather than different prognosis. An evidence-based, multicenter approach would be beneficial. PMID- 15885793 TI - Developing a model system for teaching goniotomy. AB - PURPOSE: To design a model-system instruction course to prepare trainees for performing goniotomy on patients. DESIGN: Experimental study. PARTICIPANTS: Three pediatric ophthalmology fellows and 1 recent graduate of the fellowship program. METHODS: We piloted 3 model systems: human cadaver eyes with and without Marty the Surgical Simulator (Iatrotech Inc., Del Mar, CA, hereafter referred to as Marty Head) eye supporting system and artificial eyes in Marty Head. For improving intraocular view in cadaver eyes, we used epithelial scrapping, intracameral viscoelastic, and/or intracameral lubricating jelly. Each trainee underwent a training course, including background reading, didactic lecture, and video review followed by goniotomy on model systems with the operating microscope. MAIN OUTCOME MEASURES: Each trainee evaluated each step of the course using modified 5-point Likert scales. Reading material and videos were evaluated for usefulness, readability/quality, and new information obtained. Each model system was evaluated for visibility, ease of setup, surgical feel, and transferability to live surgery. In the end, each trainee was asked to assess the overall course for usefulness. RESULTS: Trainees rated the reading materials and video highly for their usefulness and quality, yet they believed they did not learn substantial amounts of new information. Visibility and ease of setup was best with artificial eyes in the Marty Head model. Human cadaver eyes in the Marty Head provided somewhat less visibility and ease of setup, but the perceived feel and transferability to live surgery was slightly better than with artificial eyes. Cadaver eyes without the Marty Head got the lowest rating in all categories. At the end of the course, all participants felt more confident and ready to perform goniotomy on patients. All recommended this course as part of pediatric ophthalmology fellowship training programs. CONCLUSIONS: This model system instruction course can assist trainees in learning to perform goniotomy while potentially lessening the risks to patients. PMID- 15885794 TI - Ophthalmic sequelae of Crouzon syndrome. AB - PURPOSE: To document the frequency of ophthalmic sequelae in patients with Crouzon syndrome before the influence of craniofacial surgery. DESIGN: Retrospective observational case series. PARTICIPANTS: Seventy-one consecutive patients with a clinical diagnosis of Crouzon syndrome assessed before craniofacial or ophthalmic surgery at the Australian Craniofacial Unit between 1984 and 2000. METHODS: Review of clinical records with documentation of patient age, gender, visual acuity, refractive error, diagnosis of amblyopia, squint, eye movement dysfunction, nystagmus, fundus examination, examination of the anterior segment, interpupillary distance, and intercanthal distance. MAIN OUTCOME MEASURES: The frequency of ophthalmic signs and visual impairment, defined as a visual acuity of 6/12 or less. RESULTS: Visual impairment in at least 1 eye occurred in 35% of patients and was bilateral in 9%. The most common cause of visual impairment was amblyopia, which was present in 21% of patients, followed by optic atrophy in 7%. Ametropia occurred in 77% of patients; 57% had hypermetropia of > or =+2 diopters (D) and 20% had myopia of > or =-0.5 D. Strabismus occurred in 39% of patients. Although exposure keratopathy was observed in 15% of patients, this complication was well managed and caused no reduction in visual acuity. CONCLUSIONS: Early detection to reduce amblyopia by correction of refractive errors, timely treatment of strabismus, and patching should be a priority for ophthalmologists and a goal of the craniofacial teams managing patients with Crouzon syndrome. Optic atrophy remains an important cause of visual impairment in these patients before decompressive craniectomy. PMID- 15885795 TI - Noncompliance with ocular hypotensive treatment in patients with glaucoma or ocular hypertension an evidence-based review. AB - OBJECTIVE: To summarize the available scientific evidence to support clinical decisions on how to deal with noncompliance in glaucoma patients. CLINICAL RELEVANCE: Insufficient reduction of intraocular pressure and progression of visual field (VF) loss in glaucoma patients due to noncompliance with topical treatment may result in unnecessary therapy, with additional risks and costs. METHODS/LITERATURE REVIEWED: We conducted a literature search in the databases MEDLINE, EMBASE, CINAHL, PsychInfo, and Cochrane and reference lists. Thirty-four articles describing 29 original quantitative studies, in English, German, French, or Dutch, were included. Studies on noncompliance in drug trials were excluded. Two investigators independently selected the articles and abstracted their content, before negotiating their inclusion or exclusion. RESULTS: The proportions of patients who deviate from their prescribed medication regimen ranged from 5% to 80%. The impact of noncompliance on clinical outcome has not yet been established. There are no determinants sensitive and specific enough to identify potential noncompliers accurately. Patient knowledge and dose frequency can be used as starting points to improve compliance. A combination of patient education and prevention of forgetting doses seems to be successful in enhancing patient compliance. CONCLUSION: Noncompliance with hypotensive treatment is common among glaucoma patients. However, there is no strong evidence supporting a relation between noncompliance and progression of VF loss. Only a few guidelines for clinicians can be derived from the currently available literature. Future research should be guided by clinically relevant questions. PMID- 15885796 TI - Glutamate release induced by activation of glycine and GABA transporters in spinal cord is enhanced in a mouse model of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis is a progressive and fatal neurodegenerative disease, involving both upper and lower motor neurons, the cause of which is obscure, although glutamate (GLU)-induced excitotoxicity has been suggested to play a major role. We studied the release of [3H]d-aspartate ([3H]d-ASP) and endogenous glutamate evoked by glycine (GLY) or GABA from spinal cord synaptosomes in mice expressing a mutant form of human SOD1 with a Gly93Ala substitution ([SOD1-G93A(+)]), a transgenic model of amyotrophic lateral sclerosis, in mice expressing the non-mutated form of human SOD1 [SOD1+], and in non-transgenic littermates [SOD1(-)/G93A(-)]. In parallel experiments, we also studied the release of [3H]GABA evoked by GLY and that of [3H]GLY evoked by GABA. Mutant mice were killed at advanced phase of pathology or during the pre symptomatic period. In SOD1(-)/G93A(-) or SOD1(+) mice GLY evoked [3H]d-ASP and [3H]GABA release, while GABA caused [3H]d-ASP, but not [3H]GLY, release. The GLY evoked release of [3H]d-ASP, but not that of [3H]GABA, and the GABA-evoked [3H]d ASP release, but not that of [3H]GLY, were more pronounced in SOD1-G93A(+) than in SOD1(+) or SOD1(-)/G93A(-) mice. Furthermore, the excessive potentiation of [3H]d-ASP by GLY or GABA was already present in asymptomatic 30-40 day-old SOD1 G93A(+) mice. The releases of endogenous glutamate and GABA also were enhanced by GLY and the GLY-evoked release of endogenous glutamate, but not of endogenous GABA, was higher in SOD1-G93A(+) than in control animals. Potentiation of the spontaneous amino acid release is likely to be mediated by activation of a GLY or a GABA transporter, since the effect of GLY was counteracted by the GLY transporter blocker glycyldodecylamide but not by the GLY receptor antagonists strychnine and 5,7-dichlorokynurenate while the effect of GABA was diminished by the GABA transporter blocker SKF89976-A but not by the GABA receptor antagonists SR9531 and CGP52432. It is concluded that the glutamate release machinery seems excessively functional in SOD1-G93A(+) animals. PMID- 15885797 TI - Does copper ameliorate the vestibular toxicity of iminodipropionitrile (IDPN)? PMID- 15885798 TI - Imaging of cerebrospinal fluid space and movement in mice using near infrared fluorescence. AB - We developed an optical method for imaging the cerebrospinal fluid (CSF) space and the movement of CSF in mice using a near infrared fluorescence imaging methodology. Indocyanine green bound to high-density lipoprotein (ICG-HDL) was injected into the lumber subarachnoid space of nude mice. The time course of CSF movement was followed over 48 h. The imaging system was configured for epi fluorescence measurements at near infrared wavelengths using the illumination light and narrow band excitation filtration with central wavelength of 755 nm. Emission light was filtered with a long pass filter with a cutoff at 798 nm. After the injection of ICG-HDL, a strong fluorescence signal clearly delineated the cisterna magna, bilateral supracerebellar/cerebello-pontine angle cistern, and interhemispheric cistern. Much of the fluorescent tracer was washed out within 24h after the injection. This study has therefore demonstrated that an optical method, employing near infrared fluorescence imaging, can determine the CSF space and the movement of CSF in nude mice. PMID- 15885799 TI - Long-term intracellular recordings of respiratory neuronal activities in situ during eupnea, gasping and blockade of synaptic transmission. AB - For a definitive evaluation of the hypothesis that different neurophysiological mechanisms underlie the neurogenesis of eupnea and gasping, long-term continuous intracellular recordings of respiratory neuronal activities during both respiratory patterns are required. Such recordings in vivo are technically difficult, especially in small mammals, due to mechanical instability of the brainstem and cardiovascular depression that accompany hypoxia-induced gasping. Respiratory-related rhythmic activities of in vitro preparations are confounded by the lack of a clear correspondence with both eupnea and gasping. Here, we describe new methodologies and report on whole cell patch clamp recordings from the ventrolateral medulla and the hypoglossal motor nucleus in situ during multiple bouts of hypoxia-induced gasping. The longevity of recordings (range 20- 35 min) also allowed subsequent analysis of neuronal behaviour after blockade of inhibitory and excitatory synaptic activities. We conclude that whole cell patch clamp recordings in the in situ preparation will allow an analysis of both synaptic and ionic conductances of respiratory neurons during defined eupnea and gasping, providing an additional approach to in vitro preparations. PMID- 15885800 TI - A visual aid for computer-based analysis of sleep-wake state in rats. AB - Computer-based sleep scoring systems are often calibrated by reference to a conventional visual analysis of electroencephalographic (EEG) and electromyographic (EMG) traces. However, these types of data place high demands on digital storage capacity which may limit the duration or feasibility of some studies. The present paper describes an approach to visual analysis that involves reconstruction of a waveform (termed a "pseudopolygram" (PPG)) from conditioned data derived from the EEG and EMG. The PPG is the sum of three sine waves, each of which has a distinct frequency (non-REM sleep (NREM), 3 Hz; rapid eye movement sleep (REM), 7 Hz and wakefulness (WAKE), 60 Hz) and amplitude proportional to the value of a state-specific scoring variable. Thus, in NREM sleep the wave depicting the NREM quantifier has high amplitude and produces a PPG with dominant 3 Hz frequency. In REM sleep, the wave depicting the REM quantifier has high amplitude and produces a PPG with a dominant 7 Hz frequency, and in WAKE the PPG is dominated by 60 Hz. Thus, the PPG provides a means for visual discrimination of the three behavioural states. Validation studies found an overall reliability of 94% compared with conventional visual analysis of EEG and EMG. The PPG was also found to remain accurate in rats after 24 h of sleep deprivation. PMID- 15885801 TI - Evaluation of the gamma interferon test for diagnosis of paratuberculosis in goats. AB - The gamma interferon assay was evaluated for diagnosis of paratuberculosis in goats with special emphasis on false positive reactions. Four categories of herds were tested: (A) herds that had a history of paratuberculosis, had given positive Mycobacterium avium subsp. paratuberculosis fecal samples and were vaccinated against paratuberculosis; (B) herds that had been vaccinated but had never shown clinical signs of paratuberculosis nor given positive M. a. paratuberculosis fecal samples; and (C) non-vaccinated herds without paratuberculosis. To extend the analysis of samples from young goats free of paratuberculosis, animals less than 18 months of age from non-vaccinated herds without paratuberculosis, category D, were included. Heparinized blood was stimulated with purified protein derivate (PPD) from M. a. paratuberculosis for 24 h and plasma was assayed for the presence of gamma interferon. Results were recorded as the difference between OD values of PPD stimulated and control samples. Vaccinated animals from herds with paratuberculosis, category A, showed significant higher gamma interferon responses than animals from vaccinated herds without paratuberculosis, category B. In both these groups the responses were correlated to age with higher responses in younger animals. Some of the vaccinated animals in herds without paratuberculosis had a gamma interferon response lasting for several years, which demonstrate a long lasting interference with diagnostic testing in vaccinated goats. Only three of the 121 non-vaccinated animals free of paratuberculosis in category C had responses against PPD (corrected OD values at 0.2, 0.24 and 0.5), and none of the 255 young animals in category D had corrected OD values exceeding 0.2. This indicates that false positive reactions do not appear to the same extent in young goats as in young cattle. We conclude that the low responses of non-infected goats could make the gamma interferon assay useful in monitoring the paratuberculosis status of non-vaccinated herds. However, more information about the early gamma interferon responses of naturally infected goats and the presence of false negative samples are needed. PMID- 15885802 TI - Anatomical localisation and histology of the ovine tonsils. AB - The topography and histologic structure of the various tonsils were studied anatomically and microscopically in 15 sheep aged between 9 and 15 months. The palatine, pharyngeal and paraepiglottic tonsils were readily visible macroscopically. They consisted mainly of secondary lymph nodules and were encapsulated in dense connective tissues. The epithelium covering the tonsils and their crypts was frequently infiltrated heavily by lymphocytes. The tubal tonsil and the tonsil of the soft palate were macroscopically visible after fixation in 2% acetic acid. These tonsils consisted of scattered lymph nodules, aggregations of lymphocytes and diffuse lymphoid tissue. They were not encapsulated, and therefore the borders of these tonsils could not be clearly delineated. The lingual tonsil was not macroscopically visible in sheep and consisted of scattered small aggregations of lymphocytes. PMID- 15885803 TI - Antibodies against a Leishmania infantum peroxiredoxin as a possible marker for diagnosis of visceral leishmaniasis and for monitoring the efficacy of treatment. AB - Diagnosis of leishmaniasis is frequently based on serological methods, such as direct agglutination, immunofluorescence tests and ELISA assays with Leishmania total extracts, as antigen, however due to highly inconclusive results, more reliable tests are needed. In the present study, the prevalence of antibodies to a number of recombinant proteins (LmSIR2, LmS3a, LimTXNPx, LicTXNPx and LiTXN1) highly conserved among Leishmania species, were evaluated by ELISA in Leishmania infantum infected children from an endemic area of Portugal. We found that sera from children patients had antibodies against the different recombinant proteins, LicTXNPx presented the highest immuno-reactivity compared to the other and the most often recognized in the case of acute visceral leishmaniasis (VL). Moreover, in children treated with meglumine antimoniate or amphotericin B, antibodies against some of the recombinant proteins declined, whereas conventional serology using crude extracts showed little or no difference between the pre- and post treatment values. The highest reduction was observed in the case of antibodies against the LicTXNPx protein. These results suggest that the antibodies against LicTXNPx might be a useful constituent of a defined serological test for the diagnosis and the monitoring of the therapeutic response in VL. The monitoring and follow-up in a large-scale field trials of such marker in areas where leishmaniasis is endemic will lend support to this. PMID- 15885804 TI - Influence of IL-1 gene cluster polymorphisms on the development of H. pylori associated gastric ulcer. AB - BACKGROUND AND AIMS: Chronic H. pylori infection is the main cause of ulcer disease which depicts a major burden of our healthy care systems. The individual host immune response plays a pivotal role in the outcome of the infection but genetic susceptibility to develop gastric ulcer is unknown. IL-1beta and its natural receptor antagonist IL-1ra are involved in the inflammatory response to H. pylori infection. Thus, we investigated the influence of functional genetic variants in the IL-1 gene cluster on the development of gastric ulcer disease. PATIENTS AND METHODS: 390 H. pylori infected patients were genotyped for IL-1B 31 and +3954 by TaqMan technology. Alleles of IL-1RN 86VNTR were determined by gel electrophoresis after amplification. Three hundred and sixty healthy blood donors were included as healthy controls. RESULTS: Carriage of the IL-1B -31 C allele conferred a increased but not significant risk for H. pylori infection (OR: 1.3, Wald 95% CI: 0.8 2000 microg/ml for EPC cells and 345 microg/ml for RTG-2 cells), an antiviral substance was detected. Fresh filtered hemolymph was capable of inhibiting the replication of herpes simplex virus type 1 in vitro at an EC50 of 425 microg/ml (total proteins) and the replication of infectious pancreatic necrosis virus in EPC and RTG-2 cells at 217 and 156 microg/ml (total proteins), respectively. PMID- 15885818 TI - Hemispheric specialization and dual processing in strongly versus weakly lateralized chicks. AB - Lateralization of various functions is well established in the domestic chick and other vertebrates. The function of lateralization is a question under investigation now. In a previous study, it was shown that chicks incubated in the dark (Da), which prevents the development of lateralization of visual foraging and predator detection, did not perform well when two tasks requiring simultaneous use of the expertise of the right and left eye systems were presented. Performance of a task requiring the chick to find grains against a background of pebbles deteriorated in the Da chicks in the presence of a predator. In the study reported here, the two simultaneous tasks were tested in both binocular and monocular conditions to establish the role of each eye system. Learning of the pebble-floor task was poor in Da chicks tested binocularly or monocularly and in the light-exposed (Li) chicks using the left eye (LE). The time taken to complete the pebble-floor task in the presence of a predator was significantly longer for the Da chicks. This was because pecking was interrupted for longer times when the predator was presented, and the Da chicks made more distress calls than the Li. The latency to detect the predator was longest in the Li chicks using the right eye (RE) (i.e., the Li were lateralized). In the Li chicks, sustained initial viewing of the predator was by the LE. There was no LE/RE difference in the Da chicks. The intensity of responses decreased with continued presentation of the predator but forced use of the RE slowed this habituation. Exposure to light before hatching affects lateralization of both learning of the pebble-floor task and the detection of an overhead predator. Attending to the foraging and predator vigilance tasks simultaneously is impaired in the Da chicks and the superior ability of Li (lateralized) chicks is due to a specific effect of light stimulation of the RE prior to hatching. We have shown that superior performance on the dual task results from the ability to allocate food searching to one hemisphere (left) and predator vigilance to the other (right) hemisphere, achieved only by Li chicks. PMID- 15885819 TI - Effects of prenatal maternal stress by repeated cold environment on behavioral and emotional development in the rat offspring. AB - It has been reported that many types of stresses, which caused physiological and psychological alterations in dams as prenatal maternal stress, affected behavioral and emotional traits of their offspring. However, effects of environmental temperature changes, which induce various stress responses in both animals and humans, have not been assessed as prenatal maternal stress. Repeated cold stress (RCS) is a type of chronic cold stress in which environmental temperature changes rapidly and frequently several times within a day. In the present study, to investigate effects of chronic maternal stress by the RCS on behavioral and emotional development of the rat offspring (prenatal RCS rats), the RCS stress was loaded to pregnant rats between day 9 and 19 after fertilization. The prenatal RCS rats showed similar locomotor activity in an open field to control rats that were borne by non-stressed pregnant rats. On the other hand, the prenatal RCS rats showed significantly higher startle responses than the control rats in a light enhanced startle paradigm. However, treatment of diazepam decreased the startle responses in the prenatal RCS rats to the same degree as those in the control rats. The results indicated that prenatal RCS affected emotional development of the rat offspring, but not locomotor activity. Comparison of the present results with the previous studies suggests that there might be unknown common mechanisms among different prenatal maternal stresses that induce similar behavioral developmental alteration. PMID- 15885820 TI - Behavioral assessment in mouse models of neuronal ceroid lipofuscinosis using a light-cued T-maze. AB - Learning impairment is a common feature of the neuronal ceroid lipofuscinoses (NCL), a family of lysosomal storage disorders associated with progressive neurodegeneration. Murine models for the neuronal ceroid lipofuscinoses include the well-characterized motor neuron degeneration (mnd/mnd) model for one variant of late infantile NCL (CLN8), and the more recently generated models for the infantile (CLN1) and juvenile (CLN3) forms of NCL. To determine whether these mouse models exhibit behavioral deficits analogous to the learning impairment characteristic of the human disorders, the performance of these animals on an associative learning task was assessed. The abilities of affected and normal control mice to associate a light stimulus with a food reward were evaluated in 14-16-week-old animals using a T-maze. Normal mice were able to reach a criterion for having learned to make the association within a mean of 9.4 trials. The CLN8 and CLN3 mice, on the other hand, required means of 26.2 and 27.5 trials, respectively, to reach the same performance criterion (p<0.05), whereas none of the CLN1 mice were able to reach the criterion within a limit of 30 trials. The poor performance of the mutant mice did not appear to result from impaired retinal function; mice of all three strains exhibited retinal electrophysiological responses to dim light flashes and displayed robust pupillary light reflexes. Associative learning deficits appear to be an early disease phenotype in the NCL mouse models that will be useful for assessing the efficacy of therapeutic interventions such as gene or stem cell therapies. PMID- 15885821 TI - Analysis of DNA damage in sea lamprey (Petromyzon marinus) spermatozoa by UV, hydrogen peroxide, and the toxicant bisazir. AB - In this study we sought to demonstrate that Comet assay can be applied to sea lamprey sperm DNA fragmentation and used to describe the relationship between sperm DNA damage and sperm fertilizing ability. We show that the assay can be used reliably and accurately, and unlike in the case of mammals, there is no need for additional steps related to improvement of efficacy of lysis and DNA decondensation. This agrees with the presence of histone proteins in lamprey sperm. An increase in DNA fragmentation was noted during short-term storage of milt on ice (0-4 days). We demonstrated genotoxic effects of UV radiation and oxidative stress (exposure to hydrogen peroxide) and found that oxidative damage to sperm DNA was likely repaired after fertilization in the embryo. Repairing capacity of the oocyte toward sperm DNA lesions caused by UV was restricted. Toxic effect of p,p-bis-(1-aziridinyl)-N-methylphosphinothioic acid (p,p-bis(1 aziridinyl)-N-methylphosphinothioic amide), a sea lamprey chemosterilant, could not be linked to DNA fragmentation in the in vitro tests. Its genotoxicity in vivo may possibly be associated with other mechanisms of DNA degradation (oxidation or DNA-protein and DNA-DNA cross-linking). In conclusion, this study demonstrates that Comet assay can be successfully applied to monitor effects of environmental disturbances and imposed injuries in sea lamprey spermatozoa and possibly other species of ancient fish with acrosomal sperm. PMID- 15885822 TI - Mannosamine can replace glucosamine in glycosylphosphatidylinositols of Plasmodium falciparum in vitro. AB - Mannosamine (2-deoxy-2-amino-D-mannose) is unable to block GPI biosynthesis in Plasmodium falciparum: neither parasite development nor GPI biosynthesis were blocked by mannosamine treatment in P. falciparum cultures. Further, it was shown by metabolic labeling with [3H]mannosamine and subsequent monosaccharide analysis by high pH anion exchange chromatography that mannosamine is converted at a high rate into glucosamine. Both mannosamine and glucosamine are incorporated into P. falciparum glycolipids, but the characterization of mannosamine-labeled glycolipids synthesized in vivo proved difficult. Therefore, a cell-free system was developed to investigate the incorporation of [3H]mannosamine into glycolipids in P. falciparum. It was observed that mannosamine is incorporated in vitro into P. falciparum glycolipids, which possess a phosphate group. Chemical (nitrous acid deamination, mild acid hydrolysis and alkaline hydrolysis) and enzymatic (PI-PLC) treatments of [3H]mannosamine-labeled glycolipids synthesized in vitro showed the presence of GPIs. Further analyses by Bio-Gel P4 size exclusion chromatography and HPAEC demonstrated the presence of a mannosamine containing GPI-like structures, where mannosamine is incorporated instead of glucosamine, i.e. Man3-ManN-PI. This utilization of mannosamine is novel and not been described for any other cellular or parasitic system. PMID- 15885823 TI - A comparison of genetic polymorphism in populations of Onchocerca volvulus from untreated- and ivermectin-treated patients. AB - An analysis of the polymorphism of 16 genes from Onchocerca volvulus was undertaken, in two populations of worms from either ivermectin-naive patients or patients who had been repeatedly treated with ivermectin, in Ghana. Six genes were selected for analysis because studies in other nematodes had suggested a possible association with ivermectin resistance. The other 10 genes were included as control genes and have not been associated with ivermectin resistance. Twelve of the 16 genes were polymorphic, including five of the candidate genes and seven of the control genes. In all of the control genes and four of the candidate genes, there were no differences in genetic polymorphism between the untreated and ivermectin treatment worms. However, there were statically significant differences (chi2=0.05) in allelic frequencies between the untreated and treatment derived worms for P-glycoprotein and beta-tubulin genes; both genes which have been previously associated with ivermectin resistance in other nematodes. These genes were in Hardy-Weinberg equilibrium in the untreated population. However, the P-glycoprotein alleles, in the worms from the patients under treatment were not in Hardy-Weinberg equilibrium, and analysis of the allele frequencies of beta-tubulin suggested that this gene may have also been under selection in the worms from the ivermectin-treated patients. This data provides evidence of genetic selection by ivermectin on O. volvulus and indicates that investigations should be made to determine whether ivermectin resistance is developing. The beta-tubulin and P-glycoprotein genes may prove useful for monitoring for possible development of ivermectin resistance. PMID- 15885824 TI - Comment on "Correlation between the treated volume, the GTV and the CTV at the time of brachytherapy and histopathologic findings in 33 patients with operable cervix carcinoma". PMID- 15885825 TI - Radiotherapy is effective in patients with glioblastoma multiforme with a limited prognosis and in patients above 70 years of age: a retrospective single institution analysis. AB - BACKGROUND AND PURPOSE: To evaluate the efficacy of radiotherapy in patients with glioblastoma multiforme (GBM) with a limited prognosis and in patients older than 70 years. PATIENTS AND METHODS: Retrospective analysis of 202 patients with GBM treated between 1990 and 2000 in a single institution. Patients (including patients >or=70 years) were assigned to RPA groups and their survival was compared with RTOG data. RESULTS: Median survival was 8.0 months for the total group and 13.9, 10.6, 3.8, 2.1 months for RPA group III (n=17), IV (n=87), V (n=60) and VI (n=38), respectively. Median survival for patients >or=70 years was 3.6 vs. 8.1 months for 50--70 years and 11.0 months for <50. In each separate RPA group, patients >or=70 years had a similar survival compared to patients of 50- 70 years. Irradiated patients (66%) survived significantly longer than non irradiated patients: 10.6 vs. 1.9 months (P<0.0001). In RPA group V the median survival for irradiated patients was 9.4 vs. 2.1 months for non-irradiated patients. In a multivariate analysis, RT remained the only prognostic factor for survival (HR 8.9, P<0.001). CONCLUSIONS: Prognosis for patients above 70 years of age is not different from younger patients, when analyzed for separate RPA groups. For patients with a poor prognosis (i.e. RPA group V), radiotherapy improves survival significantly. PMID- 15885826 TI - A prospective study to assess the bladder distension effects on dosimetry in intracavitary brachytherapy of cervical cancer via computed tomography-assisted techniques. AB - BACKGROUND AND PURPOSE: Intracavitary brachytherapy (ICBT) is as important as external beam radiotherapy (EBRT) for the radical radiotherapy of uterine cervical cancer. The degree of urinary bladder distension during ICBT may affect the dose distribution in the bladder and rectum, to which an overdose may increase the chance of developing treatment-related complications. The purpose of this prospective study was to assess and quantify the impact of bladder distension on dosimetry in ICBT in patients with cervical cancer. PATIENTS AND METHODS: We recruited 20 patients with cervical cancer during a 12-month period. Inclusion criteria included pathological diagnosis of cervical cancer with IA to IIIB stages, and intact uterus. Patients were evaluated for brachytherapy after EBRT, and eligible individuals (cervical os could be identified clearly) were invited enter to this protocol to receive ICBT. In the first brachytherapy, bladder preparation (evacuation and distension by a Foley catheter) and CT scan were performed soon after the insertion of CT-compatible applicators. Then the bladder wall doses [median dose, maximum dose and dose-volume histograms (DVH)] were calculated via the PLATO computer planning system (Nucletron PLATO-RTS version 2.0). The individual data regarding doses and DVH were collected and compared. Bladder distension may shift the applicator position, and posterior displacement of the applicator system may increase the dose to the rectal wall, so this effect was also evaluated. RESULTS: All the continuous variables of these 20 patients followed a normal distribution. By paired t-test and multiple linear regression analysis, we found that bladder distension statistically significantly decreased the median bladder wall dose with an average reduction of 48% of the dose of an empty bladder (P<0.001), and the maximum dose did not change; on the other hand, the bladder distension did not have any adverse effects on the rectal wall doses. CONCLUSIONS: Using CT-assisted three-dimensional techniques to assess the bladder and rectal wall doses is feasible. Bladder distension reduces the median dose in the bladder wall, which may reduce treatment-related complications. PMID- 15885827 TI - Lethal pneumonitis in a phase I study of chemotherapy and IMRT for NSCLC: the need to investigate the accuracy of dose computation. PMID- 15885828 TI - Economic evaluation of proton radiation therapy in the treatment of breast cancer. AB - BACKGROUND AND PURPOSE: Proton beam therapy offers potential clinical advantages compared with conventional radiation therapy for many cancer patients. The benefits are mainly a result of a more favourable dose distribution. The treatment cost with proton radiation is higher than for conventional radiation, mainly due to the large investment cost of building a proton therapy facility. It is therefore important to evaluate whether the medical benefits of proton therapy are large enough to justify the higher treatment costs, compared with conventional radiation therapy. PATIENTS AND METHODS: The cost-effectiveness of proton therapy in the treatment of 55-year old women with left-sided breast cancer was assessed. A Markov cohort simulation model was used to simulate the life of patients diagnosed with breast cancers and treated with radiation. Cost and quality adjusted life years (QALYs) were the primary outcome measures. RESULTS: The study found a cost per QALY gained of 67,000 Euro for the base case analysis of an average breast cancer patient. The cost per QALY gained would, however, be considerably lower if a population with high-risk of developing cardiac disease was treated. Sensitivity analyses showed that the results were stable and that the risk of cardiac disease was the most important parameter. CONCLUSIONS: The results indicate that proton therapy for breast cancer can be cost-effective if appropriate risk groups are chosen as targets for the therapy. PMID- 15885829 TI - Additional dose constraints for analytical beam weighting optimization in IMRT. AB - This work presents an improvement to an algorithm for analytical beam weighting optimization where a flexible objective function, which considers 'importance factors' for each anatomical region and 'allowed deviations' from the prescribed dose, is defined. This upgrading allows forcing the mean value of the dose distribution to be the desired value, by using Lagrange multipliers. A real case is presented to show the effect of this change. PMID- 15885830 TI - Treatment planning evaluation of non-coplanar techniques for conformal radiotherapy of the prostate. AB - BACKGROUND AND PURPOSE: To evaluate the benefit of using non-coplanar treatment plans for irradiation of two different clinical treatment volumes: prostate only (PO) and the prostate plus seminal vesicles (PSV). MATERIAL AND METHODS: An inverse planning algorithm was used to produce three-field, four-field, five field and six-field non-coplanar treatment plans without intensity-modulation in ten patients. These were compared against a three-field coplanar plan. A dose of 74 Gy was prescribed to the isocentre. Plans were compared using the minimum dose to the planning target volume (PTV), maximum dose to the small bowel, and irradiated volumes of rectum, bladder and femoral head. Biological indices were also evaluated. RESULTS: For the PO group, volume of rectum irradiated to 60 Gy (V(60)) was 22.5+/-3.7% for the coplanar plan, and 21.5+/-5.3% for the five-field non-coplanar plan, which was the most beneficial (p=0.3). For the PSV group, the five-field non-coplanar plan was again the most beneficial. Rectal V(60) was in this case reduced from 41.5+/-10.4% for the coplanar plan to 35.2+/-9.3% for the non-coplanar plan (p=0.02). CONCLUSIONS: The use of non-coplanar beams in conformal prostate radiotherapy provides a small increase in rectal sparing, more significantly with PSV volumes than for PO volumes. PMID- 15885831 TI - Target delineation in post-operative radiotherapy of brain gliomas: interobserver variability and impact of image registration of MR(pre-operative) images on treatment planning CT scans. AB - BACKGROUND AND PURPOSE: To investigate the interobserver variability of intracranial tumour delineation on computed tomography (CT) scans using pre operative MR hardcopies (CT+MR(conv)) or CT-MR (pre-operative) registered images (CT+MR(matched)). PATIENTS AND METHODS: Five physicians outlined the 'initial' clinical tumour volume (CTV0) of seven patients affected by HGG and candidates for radiotherapy (RT) after radical resection. The observers performed on screen tumour delineation using post-operative CT images of the patients in the treatment position and pre-operative MR radiographs (CT+MR(conv)); they also outlined CTV0 with both CT and corresponding MR axial image on screen (CT+MR(matched)). The accuracy of the image fusion was quantitatively assessed. An analysis was conducted to assess the variability among the five observers in CT+MR(conv) and CT+MR(matched) modality. RESULTS: The registration accuracy in 3D space is always less than 3.7 mm. The concordance index was significantly better in CT+MR(matched) (47.4+/-12.4%) than in CT+MR(conv) (14.1+/-12.7%) modality (P<0.02). The intersecting volumes represent 67+/-15 and 24+/-18% of the patient mean volume for CT+MR(matched) and CT+MR(conv), respectively (P<0.02). CONCLUSIONS: The use of CT and MR registered imaging reduces interobserver variability in target volume delineation for post-operative irradiation of HGG; smaller margins around target volume could be adopted in defining irradiation technique. PMID- 15885832 TI - Comment on: "HIV infection and invasive cervical cancers, treatment with radiation therapy: toxicity and outcome" [Radiother Oncol 2005;74:31-35, Shrivastava et al.]. PMID- 15885833 TI - Effect of sesaminol glucosides on beta-amyloid-induced PC12 cell death through antioxidant mechanisms. AB - Several lines of evidence support that beta-amyloid (Abeta)-induced neurotoxicity is mediated through the generation of reactive oxygen species (ROS) and elevation of intracellular calcium. In this study, we have investigated protective effects of sesaminol glucosides on Abeta-induced oxidative cell death in cultured rat pheochromocytoma (PC12) cells. Sesaminol glucoside (50-250microg/ml) decreased Abeta(25-35)-induced ROS generation, formation of 8-oxodG, a form of oxidative DNA and elevation of intracellular calcium level concomitant with prevention of apoptotic cell death dose dependently. Sesaminol glucoside (50-250microg/ml) also effectively decreased Abeta1-42 and ADDL form of Abeta1-42 as well as the combination of H2O2 with FeSO4-induced cell damages. In mechanistic study, sesaminol glucosides attenuated Abeta25-35-induced activation of redox transcription factor nuclear factor-kappaB NF-kappaB through inhibition of p50 translocation and IkappaB phosphorylation, and blocked NF-kappaB-dependent luciferase activity in addition to the inhibitory effect on Abeta25-35-induced activation of ERK kinase signal pathway. Consistent with the inhibitory effect on Abeta25-35-induced stress-induced cell death, sesaminol glucosides decreased expression of pro-apoptotic gene p53, and Bax and caspase-3, but enhanced expression of anti-apoptotic Bcl-2. Moreover, the protective effects of sesaminol glucoside on Abeta25-35-induced ROS generation, NF-kappaB activation and cell death were further enhanced with glutathione. This study therefore suggests that sesaminol glucosides have protective effect on Abeta-induced neuronal cell death, and its effect may be through antioxidative property. PMID- 15885834 TI - Studying and modelling the combined effect of temperature and water activity on the growth rate of P. expansum. AB - The effect of solutes, water activity (a(w), 0.890--0.980) and temperature (5--25 degrees C) on the mycelial growth rate of Penicillium expansum was evaluated. The growth rate dropped as the temperature and a(w) of the medium decreased. NaCl was the solute causing the greatest growth rate reduction, followed by glucose, glycerol and sorbitol. Statistical analysis of the results showed a significant effect of solute, a(w), temperature and combinations of two or three of these factors (P<0.0001). Whatever the solutes and a(w) values, the initiation of colony growth required an additional day at 15 degrees C and 5 degrees C as compared to initiation at 25 degrees C. Growth models based on the results obtained with sorbitol and glycerol differed only slightly, with R(2) values of 97.00% and 97.95%, respectively. The response surfaces of both quadratic polynomial models showed that P. expansum should be able to grow at low a(w) (0.890) and that growth at 25 degrees C should be fastest at a(w) values ranging from 0.960 to 0.980. Both models presented a good fit between predicted and observed values. PMID- 15885835 TI - A new scaleable method for the purification of botulinum neurotoxin type E. AB - Botulinum neurotoxins belong to the most toxic substances in nature. Well-known as a food poisoning agent for almost two centuries, the beneficial aspects of this bacterial metabolite were rediscovered about 30 years ago. These toxins, which are produced by the anaerobic bacterium Clostridium botulinum are nowadays used to treat a variety of neuro-muscular disorders. The increased demand requires techniques for the production and purification of these toxins on an industrial scale. The method described herein is based on filtration and chromatography procedures only. Precipitation, centrifugation and dialysis steps were consequently excluded to develop a protocol, which can easily be scaled up from the laboratory purification to industrial needs. About 4 mg of BoNT/E were purified from a 10-L batch culture corresponding to an overall recovery of approximately 14%. PMID- 15885836 TI - Effect of re-feed strategies and non-ammoniagenic medium on adenovirus production at high cell densities. AB - Recombinant adenoviruses became one of the vectors of choice for delivery and expression of foreign proteins for gene therapy and vaccination purposes. Nevertheless, the production of adenovirus is currently limited by the so-called "cell density effect", i.e., a drop in cell specific productivity concomitant with increased cell concentration at infection (CCI). This work describes the characterisation and optimisation of the infection process in order to improve recombinant adenovirus type 5 yields at high cell densities. For that purpose, 293 cells adapted to suspension were grown in 2l bioreactors and infected at different cell concentrations, using different re-feed strategies, while evaluating cell metabolism. The consumption of amino acids is enhanced during infection, although no amino acid limitation was detected for cells infected at concentrations in the range of 2 x 10(6)cell/ml, for which the highest volumetric productivity was obtained in batch mode. Conversely, infecting at cell concentrations in the range of 3 x10(6)cell/ml led to complete depletion of glucose, glutamine and threonine before the optimal harvesting time, a significant decrease in volumetric productivity being observed; the effect of amino acids and glucose addition at infection time on cell specific and volumetric productivity of adenovirus was assessed, no improvement on adenovirus production being achieved. The effect of ammonia, present in high concentrations at 3 x10(6)cell/ml, was evaluated and seem to be detrimental; an 1.8-fold increase on adenovirus volumetric productivity was obtained for infections performed at 3 x10(6)cell/ml when non-ammoniagenic medium was used. PMID- 15885837 TI - Replication strategies of rabies virus. AB - Rabies virus (RV) is a prototype neurotropic virus that causes fatal disease in human and animals. RV infects hosts at the periphery, enters motoneurons or sensory nerves and moves to the central nervous system (CNS) via retrograde axonal transport. At later stages, there is also centrifugal spread to major exit portals, such as the salivary glands. Transmission to other hosts is facilitated by behavioral changes related to the CNS infection. Successful accomplishment of the RV infectious cycle depends on multiple functions of the virus, and of individual virus proteins, all together defining the typical pathogenicity and virulence, i.e. the biological fitness of this virus. In particular, it appears important for RV to sneak into the host without causing pronounced host responses and to preserve, at least for some time, the integrity of infected cells and of the neuronal network. The availability of reverse genetics systems that allow generation of engineered recombinant RV has provided tools for a more detailed analysis of viral functions relevant to the typical RV pathogenesis. Novel developments such as tracking of live fluorescent RV are further increasing the opportunities to decipher RV pathogenicity factors. In this review, we describe different aspects of the molecular biology of RV that are relevant to pathogenesis, with a particular emphasis on the accurate control of RV transcription, gene expression, and replication. In addition, the role of individual virus proteins in maintaining host cell integrity and supporting retrograde transport is discussed. The potential of recombinant RVs with single or multiple pathogenicity factors eliminated is being discussed in terms of vaccine and virus vector development. PMID- 15885838 TI - Borna disease virus interference with neuronal plasticity. AB - Viruses able to infect the central nervous system (CNS) are increasingly being recognized as important factors that can cause mental diseases by interfering with neuronal plasticity. The mechanisms whereby such infections disturb brain functions are beginning to emerge. Borna disease virus (BDV), which causes a persistent infection of neurons without direct cytolysis in several mammalian hosts, has recently gained interest as a unique model to study the mechanisms of viral interference with neuronal plasticity. This review will summarize several hypotheses that have been put forward to explain possible levels of BDV interference with brain function. PMID- 15885840 TI - Poliovirus and poliomyelitis: a tale of guts, brains, and an accidental event. AB - Nearly 100 years after its discovery poliovirus remains one of most thoroughly studied and best understood virus models for the molecular virologist. While poliovirus has been of vital importance for our insight into picornavirus biology at the cellular and biochemical level, it is ironic to note that, due to the early success in defeating poliomyelitis in the developed world through vaccination, many of the basic aspects of poliovirus pathogenesis remain poorly understood. This is chiefly due to the lack of an adequate and affordable animal model, save of old world monkeys. Fundamental questions, such as the identity of the target cells during the enteric phase of infection, or mechanisms of systemic spread are still unanswered. This review will attempt to summarize our current knowledge of the molecular biology of poliovirus, its pathogenesis, as well as recent advances in the areas of cell and tissue tropism and mechanisms of central nervous system invasion. PMID- 15885841 TI - Cells of the central nervous system as targets and reservoirs of the human immunodeficiency virus. AB - The availability of highly active antiretroviral therapies (HAART) has not eliminated HIV-1 infection of the central nervous system (CNS) or the occurrence of HIV-associated neurological problems. Thus, the neurobiology of HIV-1 is still an important issue. Here, we review key features of HIV-1-cell interactions in the CNS and their contributions to persistence and pathogenicity of HIV-1 in the CNS. HIV-1 invades the brain very soon after systemic infection. Various mechanisms have been proposed for HIV-1 entry into the CNS. The most favored hypothesis is the migration of infected cells across the blood-brain barrier ("Trojan horse" hypothesis). Virus production in the CNS is not apparent before the onset of AIDS, indicating that HIV-1 replication in the CNS is successfully controlled in pre-AIDS. Brain macrophages and microglia cells are the chief producers of HIV-1 in brains of individuals with AIDS. HIV-1 enters these cells by the CD4 receptor and mainly the CCR5 coreceptor. Various in vivo and cell culture studies indicate that cells of neuroectodermal origin, particularly astrocytes, may also be infected by HIV-1. These cells restrict virus production and serve as reservoirs for HIV-1. A limited number of studies suggest restricted infection of oligodendrocytes and neurons, although infection of these cells is still controversial. Entry of HIV-1 into neuroectodermal cells is independent of the CD4 receptor, and a number of different cell-surface molecules have been implicated as alternate receptors of HIV-1. HIV-1-associated injury of the CNS is believed to be caused by numerous soluble factors released by glial cells as a consequence of HIV-1 infection. These include both viral and cellular factors. Some of these factors can directly induce neuronal injury and death by interacting with receptors on neuronal membranes (neurotoxic factors). Others can activate uninfected cells to produce inflammatory and neurotoxic factors and/or promote infiltration of monocytes and T-lymphocytes, thus amplifying the deleterious effects of HIV-1 infection. CNS responses to HIV-1 infection also include mechanisms that enhance neuronal survival and strengthen crucial neuronal support functions. Future challenges will be to develop strategies to prevent HIV 1 spread in the brain, bolster intrinsic defense mechanisms of the brain and to elucidate the impact of long-term persistence of HIV-1 on CNS functions in individuals without AIDS. PMID- 15885842 TI - HIV-1 infection initiates changes in the expression of a wide array of genes in U937 promonocytes and HUT78 T cells. AB - Human monocytes/macrophages (M/M) are the major targets for human immunodeficiency virus type 1 (HIV-1) infection. To characterize the global effects of acute HIV-1 infection on gene expression in M/M, the expression levels of 550 host cell RNA transcripts in U937 human promonocytes at 2-3 days after HIV 1 infection were assessed using cDNA microarray analysis and were compared to those in the infected HUT78, a CD4+ T cell line. Confirmed by semiquantitative RT PCR, our results showed that 12 genes were up-regulated and 26 genes were down regulated in the infected U937 cells at 2-3 days post-infection, whereas 8 genes were up-regulated and 20 genes were down-regulated in the infected HUT78 cells at 2-3 days post-infection. These genes encode a host of proteins with divergent functions in a variety of cellular processes including apoptosis (FAS, Fas ligand, PIN, HSP90beta, bcl-2, bcl-x), cell signal transduction (Ras, RGS1, IRF 1, STAT3), receptor-mediated signaling transduction (CD71, CD69, CD3delta), cell cycle and growth (c-myc, cytokines, kinase), transcriptional regulation (EWS, CREB-2), and chemotaxis (beta-chemokines, RANTES), supporting the general effects of HIV-1 infection on cells of different origin. Although most identified genes were regulated similarly in both infected cell lines, differences in gene regulation, such as c-myc, CD71, CD69, and beta-chemokines, between the two infected cell lines were also identified in this study. These differences may further our understanding of the pathogenicity of HIV and enable the discovery of novel therapeutic approach for AIDS. PMID- 15885843 TI - Electroporation and transcutaneous extraction (ETE) for pharmacokinetic studies of drugs. AB - The therapeutic activity and toxicity of drugs often depends on the accumulation of drugs in the peripheral anatomical compartment rather than the central compartment. In the routine practice of therapeutic drug monitoring (TDM) and pharmacokinetic studies, drug concentration determined by intermittent blood sampling is used as a surrogate for calculating the drug concentration in the peripheral compartment tissues. Microdialysis, a relatively less invasive procedure, has been used for estimation of free drug levels in dermal, subcutaneous and muscle tissues. Transcutaneous extraction of drugs from the dermal tissue is a good noninvasive alternative to phlebotomy and microdialysis. This requires a technique, which can facilitate the extraction of significant and reproducible amounts of drugs from the dermal extracellular fluid (ECF) within a short sampling duration. In the present work, we assessed the feasibility of electroporation and transcutaneous extraction (ETE) method for determining the time course of drugs in dermal ECF, using salicylic acid (SA) as a test drug. Electroporation protocol was optimized based on the in vitro diffusion studies of salicylic acid across rat skin. The concentration-time profile of total SA was determined in rats after a single i.v. bolus administration. The in vivo permeability coefficient (P(in vivo)) of rat skin was determined under steady state plasma concentration of drug created by i.v. bolus followed by constant rate infusion of SA. The pharmacokinetic parameters of the drug were determined using a two-compartment pharmacokinetic model. The theoretical predicted time course of free SA in the dermal ECF after a single i.v. bolus administration was calculated using standard formulae. The concentration of free SA determined by ETE is in good agreement with that calculated using two-compartment pharmacokinetic model. This study thus provides a credible evidence for the validity of ETE technique for determining the concentration of SA in the dermal ECF. PMID- 15885844 TI - Polymeric gene carrier for insulin secreting cells: poly(L-lysine)-g-sulfonylurea for receptor mediated transfection. AB - Ex vivo transfer of therapeutic genes to cells is one of the potential strategies to prolong the life span of cell transplants. However, relatively safe non-viral carriers have not been extensively investigated due to their lower transfection efficiency. In this study, poly(L-lysine)-g-sulfonylurea varying SU content (PLL SU) was synthesized to promote gene delivery efficacy to an insulin secreting cell line, RINm5F, which is known to express sulfonylurea receptor (SUR). The polymer formed complexes with a model reporter gene of pCMV-Luc (DNA) and the size of resulting particles was around 100 nm. The transfection efficiency of a polymer synthesized with 5 mol% of SU in the reaction feed (PLL-SU5%) to RINm5F cell was at least 5 times higher than that of PLL. The cytotoxicity of PLL SU5%/DNA complex was equivalent to that of PLL/DNA complex. PLL-SU5% showed less transfection efficiency than PLL to NIH3T3 and HepG2 cells which are SUR negative. In RINm5F cells, the addition of free SU decreased the transfection efficiency of PLL-SU5%/DNA complex, suggesting that the complex shares the same receptors for SU. The PLL-SU5%/DNA complex seems to be internalized via SUR mediated endocytosis pathway as suggested by vacuolar ATPases inhibition by Bafilomycin A1. It is noted that RINm5F cells treated with PLL-SU5%/DNA complex secreted more insulin than control, untreated cells, suggesting the insulinotropic effect of SU in PLL-SU5%. In conclusion, PLL-SU may be useful for transfer of therapeutic genes into insulin secreting cells. PMID- 15885845 TI - Long-term (1- and 2-year) effects of lifestyle intervention in type 2 diabetes relatives. AB - AIMS: To study the long-term (1- and 2-year) effect of a lifestyle intervention on non-diabetic first-degree relatives of type 2 diabetic patients, i.e., the 1 year effect of diet versus diet and exercise in relation to a control group and the 2-year sustainability of these treatment effects. METHOD: Seventy-seven healthy first-degree relatives (men and women) between the ages of 25 and 55 were allocated to one of three groups: diet group (D), diet and exercise group (DE) and control group (C). For ethical reasons, after 1 year the control group began the intervention and were followed for another 2 years. Diet and physical activity counselling was based on current nutrition recommendations, including increased intake of fatty fish and low glycaemic index foods. The fatty acid composition of the erythrocyte membrane was studied as an objective measure of dietary change. Assessments included fasting insulin, 2-h insulin, oral glucose tolerance test (OGTT), anthropometry and blood lipid measurements. Groups D and DE received intensive follow-up through unannounced telephone interviews during the first 4 months. RESULTS: Dietary changes were significant at 1 year, and to a large degree sustained at 2 years. Adherence to advice regarding fat quality was confirmed through changes in the fatty acid composition of the erythrocyte membrane. The least active subjects in DE increased their physical activity (PA). At 1 year, group D showed a reduction in the ratio of LDL to HDL cholesterol (p=0.028) while group DE decreased their body weight by 2.7% (p<0.029) and increased HDL (p<0.037) versus controls. At 2 years, cholesterol levels (total, LDL and the ratio LDL/HDL) were reduced within group D and when compared to DE (p=0.022, 0.009, 0.035, respectively). Fasting insulin was reduced within group DE and when compared to group D (p=0.025). CONCLUSIONS: Positive changes in lifestyle, blood lipids and fasting insulin can be achieved and maintained in a non-diabetic population at risk of type 2 diabetes after 2 years. PMID- 15885846 TI - Long-term risk reduction: who needs treatment? AB - Patients with type 2 diabetes or the metabolic syndrome have an elevated risk of developing cardiovascular disease (CVD). Until recently, strategies to reduce cardiovascular risk in these patients focused mainly on controlling glycemia and blood pressure as a means of preventing disease progression. However, the metabolic abnormalities that cluster both in patients with type 2 diabetes and the metabolic syndrome are all independent risk factors for atherogenesis. There is now substantial evidence that dyslipidemia is an important, modifiable risk factor for CVD in patients with type 2 diabetes or the metabolic syndrome. Recent sub-group analyses of landmark statin trials, such as the Heart Protection Study (HPS), confirm that the benefits of intensive statin therapy on CVD risk extend to patients with type 2 diabetes, irrespective of baseline lipid levels. In addition, the Collaborative AtoRvastatin Diabetes Study (CARDS), the first statin study to focus solely on patients with type 2 diabetes, was stopped early due to the overwhelming benefits of statin therapy on cardiovascular risk in a study population who previously would not have been considered dyslipidemic (mean baseline low-density lipoprotein cholesterol [LDL-C], 3.0 mmol/l [116 mg/dl]). As a result, treatment guidelines are setting increasingly stringent goals for LDL-C levels in an attempt to reduce cardiovascular risk. However, with physicians estimating that approximately 50% of patients with type 2 diabetes do not achieve these goals, initiation of appropriate, effective and rapid-acting statin therapy is paramount in these high-risk patients. PMID- 15885847 TI - Time costs of waiting, doctor-shopping and private-public sector imbalance: microdata evidence from Hong Kong. AB - OBJECTIVE: To quantify the disutility of waiting for a specialist consultation and to examine whether individuals' willingness-to-pay (WTP) to reduce waiting time is associated with doctor-shopping behavior in outpatient clinics of four large, public hospitals in Hong Kong. METHODS: We performed a cross-sectional survey of 6495 outpatients on public sector waiting lists enrolled from July 2000 to October 2001. Main outcome measures were WTP to reduce queue time and odds ratios (ORs) for doctor-shopping according to WTP status. RESULTS: The majority (74.5%) of respondents, excluding those registering "protest zeros", was willing to pay a median of 100HK Dollar (13US Dollar ) to reduce queue time by 2 weeks, which represents 14.6% of the total costs of care for a specialist outpatient visit. Respondents' WTP decreased as their expected waiting period increased. Multivariate regression modeling showed that those who were willing to pay more were also more likely to "doctor-shop" (ORs and 95% confidence intervals for doctor-shopping for the 1st through 4th quartile of WTP values respectively = 1.00; 0.88 (0.63, 1.23); 1.19 (0.57, 2.48); 1.52 (1.05, 2.20); p for linear trend = 0.009), in effect reducing waiting time to see a specialist by seeking and paying for care in the private sector. CONCLUSIONS: Based on this preliminary evidence, we propose the further examination of a new policy of outsourcing some specialist care services to the private sector to improve allocative and technical efficiency. PMID- 15885848 TI - Automatic extraction of brain surface and mid-sagittal plane from PET images applying deformable models. AB - In this study, we propose and evaluate new methods for automatic extraction of the brain surface and the mid-sagittal plane from functional positron emission tomography (PET) images. Designing methods for these segmentation tasks is challenging because the spatial distribution of intensity values in a PET image depends on the applied radiopharmaceutical and the contrast to noise ratio in a PET image is typically low. We extracted the brain surface with a deformable model which is based on a global optimization algorithm. The global optimization allows reliable automation of the extraction task. Based on the extracted brain surface, the mid-sagittal plane was determined. The method was tested with the image of the Hoffman brain phantom (FDG) and the images from the brain studies with the FDG (17 images) and the C11-Raclopride tracers (4 images). In addition to the brain surfaces, we applied the deformable model for extraction of the coarse cortical structure based on the tracer uptake from FDG-PET brain images. The proposed segmentation methods provide a promising direction for automatic processing and analysis of PET brain images. PMID- 15885849 TI - Modified minimal model using a single-step fitting process for the intravenous glucose tolerance test in Type 2 diabetes and healthy humans. AB - The classical minimal model with single compartment was modified by the assumption that the insulin decay rate is not always a first-order process, and a mathematical function for describing the insulin infusion rate is introduced. The modified model was used to study four sets of published data including healthy humans and Type 2 diabetes with different types of insulin infusion rates. The single-step fitting process took the glucose-insulin system as a dynamic integrated physiological system and generated the real optimized model parameters from the experimental data using the modified model. It also avoided the errors from the interpolation or extrapolation for taking measured insulin points as inputs, which were mostly employed in publications when using the single or multi compartments minimal model. The averaged R(2) value between measured and calculated plasma concentrations for these four cases is 0.977, which indicates excellent agreement. PMID- 15885850 TI - Pharmaceutical applications of Mid-IR and Raman spectroscopy. AB - Mid-IR and Raman spectroscopy are versatile tools in pharmaceutics and biopharmaceutics, with a wide field of applications ranging from characterization of drug formulations to elucidation of kinetic processes in drug delivery. After an introduction to the basic principles of IR and Raman spectroscopy, new developments in applications of these methods for studying drug delivery systems, in particular topical drug delivery, will be reviewed. FTIR-ATR is a well established standard method used to study drug release in semisolid formulations, drug penetration, and influence of penetration modifiers; it is also capable of in vivo studies. FTIR-PAS has been applied to measure drug content in semisolid and solid formulations, to determine drug penetration into artificial and biological membranes. The big advantage of this technique is the possibility of spectral depth profiling. However, FTIR-PAS is so far limited to in vitro investigations. Raman spectroscopy can be used to characterize the structure of colloidal drug carrier systems. Raman spectroscopy is readily applicable to in vivo studies, but such investigations must fulfill the relevant laser safety guideline. Recently, there has been tremendous technical improvement in vibrational microspectroscopy. FTIR imaging shows great promise in its ability to visualize the drug and excipient distribution in pharmaceutical formulations such as tablets and therapeutic transdermal systems, as well as to reveal the mechanism of drug release. Furthermore, this unique technique offers completely new possibilities to study the lateral diffusion of drugs. PMID- 15885851 TI - Natriuretic peptides and their receptors. PMID- 15885852 TI - Antimicrobial and cytolytic properties of the frog skin peptide, kassinatuerin-1 and its L- and D-lysine-substituted derivatives. AB - Kassinatuerin-1, a 21-amino-acid C-terminally alpha-amidated peptide first isolated from the skin of the African frog Kassina senegalensis, adopts an amphipathic alpha-helical conformation in a membrane-mimetic solvent (50% trifluoroethanol) and shows broad-spectrum antimicrobial activity. However, its therapeutic potential is limited by its relatively high cytolytic activity against mammalian cells. The antimicrobial and cytolytic properties of a peptide are determined by an interaction between cationicity, hydrophobicity, alpha helicity and amphipathicity. Replacement of the C-terminal alpha-amide group in kassinatuerin-1 by carboxylic acid decreased both cationicity and alpha-helicity, resulting in an analog with decreased potency against Escherichia coli (4-fold) and Staphylococcus aureus (16-fold). Low cytolytic activities against human erythrocytes (LD50>400 microM) and L929 fibroblasts (LD50=105 microM) were also observed. Increasing cationicity, while maintaining amphipathic alpha-helical character, by progressively substituting Gly7, Ser18, and Asp19 on the hydrophilic face of the alpha-helix with L-lysine, increased antimicrobial potency against S. aureus and Candida albicans (up to 4-fold) but also increased hemolytic and cytolytic activities. In contrast, analogs with d-lysine at positions 7, 18 and 19 retained activity against Gram-negative bacteria but displayed reduced hemolytic and cytolytic activities. For example, the carboxylic acid derivative of [D-Lys7, D-Lys18, D-Lys19]kassinatuerin-1 was active (minimum inhibitory concentration (MIC)=6-12.5 microM) against a range of strongly antibiotic-resistant strains of E. coli but showed no detectable hemolytic activity at 400 microM and was 4-fold less cytolyic than kassinatuerin-1. However, the reduction in alpha-helicity produced by the D-amino acid substitutions resulted in analogs with reduced potencies against Gram-positive bacteria and against C. albicans. PMID- 15885853 TI - Aminopeptidase N/CD13 targeting fluorescent probes: synthesis and application to tumor cell imaging. AB - A family of fluorescein-peptide conjugates (CNP1-3) for aminopeptidase N (APN/CD13) targeting fluorescent probes were designed and synthesized. Among the three conjugates, CNP1 bearing tumor-homing cyclic peptide CNGRC, could selectively label APN/CD13 over-expressing on the surface of tumor cells of HT 1080, as identified by means of fluorescent microscopic cell imaging. CNP1 was shown to be a promising fluorescent probe applicable to tumor-targeting molecular imaging. PMID- 15885854 TI - CD40-CD40L interactions promote neuronal death in a model of neurodegeneration due to mild impairment of oxidative metabolism. AB - Abnormalities in oxidative processes, region-selective neuron loss, inflammation and diminished activity of thiamine-dependent enzymes characterize age-related neurodegenerative diseases. Thiamine deficiency (TD) models the selective neurodegeneration that accompanies mild impairment of oxidative metabolism. As in human neurodegenerative diseases, alterations in multiple cell types accompany the TD-induced neurodegeneration. The current studies demonstrate that CD40 and CD40 ligand (CD40L), two co-stimulatory immune molecules, are involved in TD induced selective neuronal death. TD induced CD40 immunoreactivity in microglia and CD40L immunoreactivity in astrocytes. Both CD40-positive microglia and CD40L positive astrocytes increased during the progressive TD-induced neuronal death. In early stages of TD, targeted deletion of CD40 diminished the number of CD40L positive astrocytes and reduced neuronal death by 35%. The number of CD40L positive astrocytes increased whenever the number of NeuN-positive neurons decreased. In early stages of TD, deletion of CD40L diminished CD40-positive microglia and reduced the neuronal death by 64%. In advanced phases of TD, neither CD40 nor CD40L deletion protected against neuronal death. The data show for the first time that TD induces expression of CD40 by the microglia and CD40L by astrocytes. The results indicate that CD40-CD40L interactions promote neuronal death in early stages of TD, but that at later phases the protective effects of the diminished CD40 or CD40L are over-ridden by other mechanisms. PMID- 15885855 TI - [The establishment of geriatric intervention group and geriatric assessment at emergency of Henri-Mondor hospital]. AB - BACKGROUND: The official French demographic previsions are a growing number of the older than 75 years elderly people. The Emergency services face this demographic evolution. We describe the establishment of the geriatric intervention group at the emergency of the Henri-Mondor university hospital at Creteil (France) and analyse the results of the geriatric assessment at the short unit care during the first four months. METHODS: We analysed the results of the geriatric assessment of 206 patients during the first four months, by considering the final unit care. The geriatric assessment evaluates functional abilities, cognitive status and thymic function with elderly people validated tests and subjective assessment of nutrition status and the sensorial functions. RESULTS: The statistical analysis of the geriatric assessment results was significant among the different hospitalized groups of patients, for the cognitive status, the nutritional risk and the walk and standing evaluation. CONCLUSION: The results of the geriatric assessment at emergency showed cognitive impairment and gait abnormality in elderly patients were at risk of hospitalization. PMID- 15885856 TI - Determining student satisfaction in a nursing college. AB - This study was carried out to determine the satisfaction and expectations of students in a nursing college in Turkey and is descriptive and prospective in nature. A student satisfaction scale developed by the researchers was used in this study. With no method of sampling employed, a total of 694 students attending the nursing college during the years of 1999, 2000 and 2001 were included in the study. The results showed that student satisfaction increased in 2001. Mean satisfaction score was high in the first year (x = 126) but decreased in the second year (x = 98). The third year was when the score reached its highest level (x = 128), and the fourth year was when it decreased to its lowest level (x = 95). The students included in this study strongly believed that there was room for improvement in the present education system, recreational activities and other services rendered by their college. Most of the students felt that the nursing college they were attending fell short of their expectations. Of note, many of them declared to have started nursing education as a last resort, that it was not their first choice, and that they did not score high enough in the university entrance exam to enable them to study a different subject. PMID- 15885857 TI - Palliative care for end-stage dementia: a discussion of the implications for education of health care professionals. AB - The increasing burden of chronic disease demands that palliative care clinicians address the needs of patients with non-malignant disease. This discussion document seeks to address some of the challenges to providing palliative care for end-stage dementia (ESD) and the need for skill enhancement in key providers of care. In spite of the intent, there is an apparent lack of appropriate, co ordinated and comprehensive palliative care available for these individuals and their families. There is an absence of well-articulated models to assist health care providers of ESD clients. It would appear that the development and evaluation of guidelines, implementation of education programs and collaborative associations between palliative and aged-care providers of care are key strategies to facilitate palliative care for ESD clients. PMID- 15885858 TI - (210)Pb content in natural gas pipeline residues ("black-powder") and its correlation with the chemical composition. AB - The present work was carried out to assess the (210)Pb content in "black-powder" found in pigging operations on gas pipelines in Brazil, in particular, on the Campos Basin gas pipeline. Additionally, the chemical composition of such deposits was determined and an eventual correlation with (210)Pb concentration evaluated. Typical "black-powder" generated in the natural gas pipeline from Campos Basin oilfield contains mainly iron oxide ( approximately 81%) and residual organic matter ( approximately 9%). The (210)Pb content ranges from 4.9 to 0.04k Bqkg(-1) and seems to be inversely correlated with the distance to the platforms. On the other hand, (226)Ra concentration is higher on the pipeline branch between the platform and the onshore installations. (228)Ra was only observed in few samples, in particular, in the samples with the highest (226)Ra content. PMID- 15885859 TI - Scapular kinematics during humeral elevation in adults and children. AB - BACKGROUND: Appropriate motion of the scapula is important for dynamic positioning of the glenoid during humeral elevation. A number of studies have described the typical scapular kinematics during humeral elevation in adults. However, children and adults may have differences in scapulothoracic musculature and scapular osteology. To our knowledge, no study has been performed examining scapular kinematics in children with either typical or atypical development. Consequently in children the influence of age and development on scapular motion is currently unknown. The aim of this study was to describe and compare the kinematic patterns of the scapula during humeral elevation in children with typical development and healthy adults. METHODS: Fifteen adults, 7 females, 25-37 years of age, and 14 children, 8 females, 4-9 years of age, participated in this study. Kinematic data were collected using a magnetic tracking device. Subjects were asked to elevate their arm in the scapular plane (40 degrees anterior to the frontal plane) in a sequence of three trials. FINDINGS: Significant differences were seen between the two age groups in the dependent variables. During scapular plane rotation from 25 degrees to 125 degrees , children showed greater upward rotation (43.9 degrees SD 6.39 degrees ) than adults (29.1 degrees SD 10.1 degrees ). The mean glenohumeral to scapulothoracic ratio in the scapular plane was 2.4:1 for adults, 1.3:1 for children. INTERPRETATION: This study demonstrates that there are significant differences in scapular kinematic patterns between children and adults. Children have a greater contribution from the scapulothoracic joint, specifically upward rotation toward humeral elevation. From a clinical perspective, these results can be used to help determine the incorporation of stabilization and mobilization of the scapulothoracic joint during exercises for a child with impairment at the shoulder for improving shoulder function. PMID- 15885860 TI - Myelodysplasic syndromes: a comprehensive review. AB - Myelodysplastic syndromes (MDS) are a set of oligoclonal disorders of hematopoietic stem cells characterized by ineffective hematopoiesis that manifest clinically as anemia, neutropenia, and/or thrombocytopenia of variable severity. The result often is transfusion-dependent anemia, an increased risk of infection or hemorrhage, and a potential to progress to acute myelogenous leukemia (AML). Although progression to acute leukemia can lead to death in patients with MDS, many deaths are consequences of cytopenias and marrow failure in the absence of transformation. Approximately 2/3 of patients succumb to the disease within 3-4 years after presentation, and individuals with high-risk MDS generally survive about 1 year. Given that the disease is more prevalent in the elderly who often have comorbid conditions, the current treatment of MDS consists mainly of supportive care. Curative treatments are restricted to younger, healthy individuals with histocompatible (HLA)-matched donors for allogenic transplant or those able to undergo intensive chemotherapeutic regimens. However, understanding of the pathophysiology of MDS and identification of potential cellular and molecular targets in recent years has led to novel therapeutic approaches. Encouraging results using these heterogeneous therapeutic approaches alone or in combination in Phase I and II trials, have, in turn, called into question previous classification systems and have confirmed the need for an all encompassing molecular, diagnostic and prognostic staging system. PMID- 15885861 TI - Selenite reduction in Boom clay: Effect of FeS(2), clay minerals and dissolved organic matter. AB - Several experiments were set up to study Se speciation and solubility in the reducing Boom clay environment, starting from oxidized Se species which were added in oversaturation with respect to the thermodynamic solubility of reduced Se solid phases. Upon introduction of SeO3(2-) to FeS(2)-containing samples, adsorption of SeO3(2-) occurred at the FeS2 surface, and led to a reduction and precipitation of a Se0 solid phase with a solubility of 3x10(-9) M (after 60 days). In the presence of humic substances, an association of Se with these humic substances was observed and the 3x10(-9) M solubility limit was not reached in the same time delay. Upon introduction of SeO3(2-) to Boom clay suspensions (equilibration up to 9 months), the initial adsorption of SeO3(2-) on the solid phase was increased with respect to systems containing only FeS2, due to the presence of (illite) clay minerals. This competing adsorption process, and the presence of humic substances, again decreased the kinetics of reduction with respect to FeS2 samples. Also, an association of Se with Boom clay humic substances was observed, and amounted up to approximately 10(-7) M in some samples after 9 months equilibration. PMID- 15885862 TI - Mercury loss from soils following conversion from forest to pasture in Rondonia, Western Amazon, Brazil. AB - This work reports on the effect of land use change on Hg distribution in Amazon soils. It provides a comparison among Hg concentrations and distribution along soil profiles under different land use categories; primary tropical forest, slashed forest prior to burning, a 1-year silviculture plot planted after 4 years of forest removal and a 5-year-old pasture plot. Mercury concentrations were highest in deeper (60-80 cm) layers in all four plots. Forest soils showed the highest Hg concentrations, ranging from 128 ngg(-1) at the soil surface to 150 ngg(-1) at 60-80 cm of depth. Lower concentrations were found in pasture soils, ranging from 69 ngg(-1) at the topsoil to 135 ngg(-1) at 60-80 cm of depth. Slashed and silviculture soils showed intermediate concentrations. Differences among plots of different soil-use categories decreased with soil depth, being non significant below 60 cm of depth. Mercury burdens were only statistically significantly different between pasture and forest soils at the topsoil, due to the large variability of concentrations. Consequently, estimated Hg losses were only significant between these two land use categories, and only for the surface layers. Estimated Hg loss due to forest conversion to pasture ranged from 8.5 mgm(-2) to 18.5 mgm(-2), for the first 20 cm of the soil profile. Mercury loss was comparable to loss rates estimated for other Amazon sites and seems to be directly related to Hg concentrations present in soils. PMID- 15885863 TI - Terrestrial avoidance behaviour tests as screening tool to assess soil contamination. AB - To assess soil quality and risk assessment, bioassays can be useful tools to gauge the potential toxicity of contaminants focusing on their bioavailable fraction. A rapid and sublethal avoidance behaviour test was used as a screening tool with the earthworm Eisenia andrei and the isopod Porcellionides pruinosus, where organisms were exposed during 48 h to several chemicals (lindane, dimethoate and copper sulphate, for isopods and carbendazim, benomyl, dimethoate and copper sulphate for earthworms). Both species were also exposed to soils from an abandoned mine. For all bioassays a statistical approach was used to derive EC50 values. Isopods and earthworms were able to perceive the presence of toxic compounds and escaping from contaminated to clean soil. Furthermore the behaviour parameter was equally or more sensitive then other sublethal parameters (e.g. reproduction or growth), expressing the advantages of Avoidance Behaviour Tests as screening tools in ERA. PMID- 15885864 TI - Obsessive compulsive disorder: a review of possible specific internal representations within a broader cognitive theory. AB - Obsessive Compulsive Disorder (OCD) is one of the most incapacitating of anxiety disorders, and is rated as a leading cause of disability by the World Health Organization (1996). Current cognitive models of OCD have focused on beliefs and management strategies involved in the development, maintenance, and exacerbation of OCD. However, despite evidence of their association to psychopathology, few researchers have applied the idea of underlying cognitive-affective structures, such as perceptions about the self and world, as operating in individuals with obsessive-compulsive thoughts and behaviors. This paper critically engages with current cognitive, developmental, and attachment research associated with views about the self and world. It is argued that consideration of such underlying cognitive-affective vulnerabilities may lead to a broader understanding of the development and maintenance of OCD. Consistent with previous theoretical work (e.g. ), we also argue that early experiences of parenting lead to the development of a dysfunctional self-structure and world-view relevant to OCD. Thus, this paper aims to extend the focus of current OCD research by exploring the possible role of a broader range of underlying vulnerability structures in the development and maintenance of OCD-related dysfunctional beliefs and symptoms. PMID- 15885865 TI - The presence of drug in control samples during toxicokinetic investigations--a Novartis perspective. AB - During a submission procedure, the validity of a few dietary toxicity studies was questioned because low levels of the drug were detected among control toxicokinetic samples. Although several lines of reasoning suggested that these findings arose from ex vivo contamination, the Regulatory Authority stated that it was not possible to establish a no-effect-level in any of the studies and so the submission was withdrawn. In response, Novartis conducted a thorough review and modification of the procedures involved in the collection and analysis of toxicokinetic samples to minimize such contamination in future studies. Ongoing monitoring of contamination in toxicology studies has subsequently revealed that although it was not possible to completely eliminate the problem, the new procedures together with an increasing awareness of the issue have considerably reduced the incidence of contamination. The process of contamination and its control was also modeled in a feeding study in mice. This provided good evidence that the detection of drug in control samples in the previous studies originated from external sources and not from in vivo exposure. PMID- 15885866 TI - Estimation of private and social time preferences for health in northern Tanzania. AB - Time preferences for health has a role in informing discounting practice in economic evaluation of health care interventions. Yet virtually no empirical evidence on time preferences for health are available for sub-Saharan Africa. The objectives of this paper are therefore to estimate time preferences for health in Tanzania, and to explore the relationship between time preferences for one's own private health and for others health. Determinants of time preference rates are also explored. A sample of the general population participated, and each person was asked to indicate the maximum duration of more distant ill-health they would be willing to accept in return for a specified delay in the onset of the condition. People were randomised to answer either questions in the context of their own health or in the context of others health. Estimations were done using clustered least squares regression with robust standard errors. The implied mean and median discount rates were 0.07 and 0.058, respectively. Time preferences for one's own and others' health--conceptualised as social health--seem to be roughly the same in this sample, but people seem to discount the most severe health condition at a significantly lower rate than the least severe condition. PMID- 15885867 TI - Subchronic and chronic safety studies with genistein in dogs. AB - Genistein is a phytoestrogen that occurs naturally in the diet, especially in soy based foods. There is widespread interest in phytoestrogens as chemopreventive agents for a variety of diseases and cancers based on epidemiologic evidence. Although soy and its constituents, such as genistein, have been consumed at high levels in several Asian populations without apparent adverse effects, concern has been raised about potential adverse effects due to estrogenic and other activities. The subchronic and chronic safety of genistein were evaluated in the beagle dog including a 4-week study and a 52-week safety study with a 13 week interim sacrifice and a 4 week recovery period. In both studies at doses of 50, 150 and 500 mg/kg/day, genistein was well tolerated. In the 4 week study, except for an increase in uterine weights in female dogs at 500 mg/kg/day, there were no other treatment related findings. In the 52-week study, the primary effects of genistein were observed on the reproductive tract, which included for male dogs: reduced size and/or weight of the testes, epididymus and prostate of 2/2 dogs after 13 weeks of treatment and in 1/4 dogs after 52 weeks of treatment at 500 mg/kg/day. The histological changes observed in the affected dogs at 500 mg/kg/day indicated atrophy of the testes and prostate gland and absent spermatozoa in the epididymus. At the mid-dose of 150 mg/kg/day, although there was a reduction to a lesser extent in testes weight after 13, but not 52 weeks, there were no histopathological changes. In female dogs, the reproductive tract effects included increased uterine weight at 500 mg/kg/day after 13 weeks of treatment, but not after 52 weeks of treatment. There was also a small decrease in ovarian weights at 150 and 500 mg/kg/day after 13 weeks and at 500 mg/kg/day after 52 weeks of treatment. There were no histopathological correlates to the changes in organ weights in female dogs. In the 4-week recovery group dogs, no changes were observed in dogs previously treated for 52 weeks with 500 mg/kg/day of genistein. It is concluded that the administration of genistein to dogs for a period of 4-52 weeks was well tolerated and did not result in systemic toxicity. Effects of genistein on the reproductive tract at very high doses were functional in nature and are of a type that would be expected in view of the relatively weak estrogenic activity of genistein and were considered not adverse effects. In the 4-week study, the no observed adverse effect level (NOAEL) for genistein was considered to be >500 mg/kg/day and the no observed effect level (NOEL) was considered to be 150 mg/kg/day. For the 52-week study, the NOAEL is considered to be >500 mg/kg/day and the NOEL is considered to be 50 mg/kg/day. PMID- 15885868 TI - Kinetic analysis of cytokine response to cigarette smoke condensate by human endothelial and monocytic cells. AB - Atherosclerosis is generally considered an inflammatory disease characterized by the accumulation of lipid in large and medium elastic arteries. Individuals who smoke are at increased risk for developing atherosclerosis and the clinical events associated with this disease. Underlying the mechanisms involved in atherosclerotic lesion development exists a complex pattern of signaling, involving molecules (cytokines and chemokines) that mediate the progression of arterial lesions. The unique nature of exposure to tobacco-related toxicants during the process of smoking prompted our investigation of the time-dependent responses of two critical cell types to cigarette smoke condensate exposure. In this study, we examined the kinetic responses, using suspension array technology and RT-PCR of 17 cytokines (IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17 GM-CSF, G-CSF, INF-gamma, TNF-alpha, MCP-1 and MIP-1beta) in human aortic endothelial cells (HAECs) and THP-1 monocyte macrophages following exposure to cigarette smoke condensate (CSC) for 24h. In HAECs, IL-8 and IL-4 were rapidly stimulated by CSC exposure while, surprisingly, MCP-1 expression was downregulated. In THP-1 macrophages, IL-6, MIP-1beta, MCP-1 and IL-1beta protein expression were suppressed upon CSC exposure. All other measurable cytokines in THP-1 cells exposed to CSC had levels of protein and mRNA similar to controls. Depending on cell type, CSC uniquely influences the expression of cytokines. The complex interplay of these signaling molecules within the framework of atherosclerosis points to the ability of cigarette smoke components to alter such signaling following acute exposure, and by this mechanism may alter the course of both atherogenesis initiation and progression. PMID- 15885869 TI - Stabilization of purine motif DNA triplex by a tetrapeptide from the binding domain of HMGBI protein. AB - DNA triple helix has attracted a lot of attention in the recent past because of their potential therapeutic applications to inhibit the transcription of genes associated with several human diseases. Therefore, studies on DNA triplex formation and stabilization have become very essential to understand the nature of DNA triplex both in quantitative and qualitative terms. We have studied the effect of a tetrapeptide, Pro-Lys-Arg-Trp (PKRW), containing the conserved sequence (PKR) of high mobility group (HMG) B1 DNA binding domain on the stability of purine motif DNA triplex. The triple stranded DNA complex (23R*R*Y) containing (GAA) repeats used in the present study is formed by: 5' TCGCGAAGAAGAAGAAGAACGCT-3', 5'-AGCGTTCTTCTTCTTCTTCGCGA-3' and the third strand 5' AAGAAGAAGAAGAAG-3'. The DNA triplex 23R*R*Y and its complex with PKRW were investigated by DNA-melting, UV-absorption, circular dichroism and fluorescence spectroscopy. The increased stability of 23R*R*Y triplex in presence of PKRW is supported by the following observations; (i) a marked increase in triplex melting temperature (ii) significant increase in the ellipticity of the characteristic negative band at 210 nm and (iii) a large change in free energy, Delta DeltaG (DeltaG of triplex-peptide complex - DeltaG of free triplex), -9.6 kcal mol(-1) on complexation with PKRW. Based on the above observations and fluorescence quenching data, we suggest a two stage model for the binding of PKRW to 23R*R*Y in which (a) PKRW binds to the minor groove of the 23R*R*Y with lysine and arginine making electrostatic interactions with phosphates of DNA, which is followed by (b) intercalation of tryptophan side chain between the A*A*T base triplets. We conclude that PKRW preferentially stabilizes Hoogsteen base pair to the Watson-Crick base pair in purine motif 23R*R*Y DNA triplex. It is tempting to speculate that HMGB1 plays an important role in recognizing the DNA triplex structures like H-DNA in the gene, which may have biological implications with respect to chromatin structure and its function in the cell. PMID- 15885870 TI - Fate of mRNA extremities generated by intrinsic termination: detailed analysis of reactions catalyzed by ribonuclease II and poly(A) polymerase. AB - In all living cells 3' ends of RNA are posttranscriptionally elongated or shortened by nucleotidyl transferases and ribonucleases. The detailed analysis of the rpsO mRNA of Escherichia coli presented here demonstrates that transcription terminates in vivo at two sites located seven and eight nucleotides downstream from the GC-rich hairpin of the intrinsic terminator and that primary transcripts can be shortened by RNase II. The shortest RNA identified in the cell result from nibbling of primary transcripts. Primary transcripts and nibbled molecules can also be adenylated by poly(A) polymerase I (PAP I). In addition, kinetics of decay performed in vitro demonstrate that RNase II rapidly degrades poly(A) tails longer than 7-8 As processively while it slowly nibbles shorter tails and non adenylated RNAs distributively. Comparison of the kinetics of nibbling of oligoadenylated rpsO mRNA in vivo and in vitro lead us to conclude that the rates of shortening and elongation of the oligo(A) tails detected in vivo are very slow: about 0.5-7 nucleotides per min. We finally speculate that the slowness of oligo(A) synthesis may explain why polyadenylation does not affect the stability of mRNAs whose degradation is controlled by RNase E. PMID- 15885871 TI - A chymotrypsin-like proteinase from the midgut of Tenebrio molitor larvae. AB - A chymotrypsin-like proteinase was isolated from the posterior midgut of larvae of the yellow mealworm, Tenebrio molitor, by ion-exchange and gel filtration chromatography. The enzyme, TmC1, was purified to homogeneity as determined by SDS-PAGE and postelectrophoretic activity detection. TmC1 had a molecular mass of 23.0 kDa, pI of 8.4, a pH optimum of 9.5, and the optimal temperature for activity was 51 degrees C. The proteinase displayed high stability at temperatures below 43 degrees C and in the pH range 6.5-11.2, which is inclusive of the pH of the posterior and middle midgut. The enzyme hydrolyzed long chymotrypsin peptide substrates SucAAPFpNA, SucAAPLpNA and GlpAALpNA and did not hydrolyze short chymotrypsin substrates. Kinetic parameters of the enzymatic reaction demonstrated that the best substrate was SucAAPFpNA, with k(cat app) 36.5 s(-1) and K(m) 1.59 mM. However, the enzyme had a lower K(m) for SucAAPLpNA, 0.5 mM. Phenylmethylsulfonyl fluoride (PMSF) was an effective inhibitor of TmC1, and the proteinase was not inhibited by either tosyl-l-phenylalanine chloromethyl ketone (TPCK) or N(alpha)-tosyl-l-lysine chloromethyl ketone (TLCK). However, the activity of TmC1 was reduced with sulfhydryl reagents. Several plant and insect proteinaceous proteinase inhibitors were active against the purified enzyme, the most effective being Kunitz soybean trypsin inhibitor (STI). The N-terminal sequence of the enzyme was IISGSAASKGQFPWQ, which was up to 67% similar to other insect chymotrypsin-like proteinases and 47% similar to mammalian chymotrypsin A. The amino acid composition of TmC1 differed significantly from previously isolated T. molitor enzymes. PMID- 15885872 TI - Collagen-induced differential expression of an RNA polymerase subunit by breast cancer cells. AB - It was previously reported that the stroma of ductal infiltrating carcinoma (DIC) of the human breast contains considerable amount of an embryo-foetal collagen type, OF/LB (onco-foetal/laminin-binding), and that adhesion of 8701-BC DIC cells onto OF/LB collagen substrates selectively promotes cell growth, motility, production of extracellular lytic enzymes and invasion "in vitro" if compared with other collagen species. To detect possible transcriptional differences for regulatory proteins following OF/LB collagen-cell interactions, we submitted RNA preparations from 8701-BC cells grown on collagen type I, IV and OF/LB to "differential display"-PCR in the presence of degenerate C(2)H(2) zinc finger and protein tyrosine kinase domain oligonucleotide primers. We report that growth of 8701-BC cells on OF/LB collagen is consistently associated with the up-regulation of hRPB17 gene, coding for an RNA polymerase subunit, as confirmed by conventional RT-PCR and Northern analyses. PMID- 15885873 TI - Purification, characterization and enzymatic degradation of YCP, a polysaccharide from marine filamentous fungus Phoma herbarum YS4108. AB - YCP, a mitogenic polysaccharide with its molecular weight (MW) of 2.4 x 10(3) kDa, was isolated from the mycelium of the marine filamentous fungus Phoma herbarum YS4108 by a combination of ion-exchange chromatography on DEAE-32 and gel permeation over Sephacryl S-400. The detailed compositional, spectroscopic and methylation analyses of the polysaccharide demonstrated that its backbone possessed most likely a linear alpha-(1 --> 4) bonded glucopyranoside main chain co-bearing through side alpha-(1 --> 6)-linkage. The alpha-(1 --> 4) bondage of the glucopyranoside building blocks in YCP was confirmed by the observation that it could be hydrolyzed by the alpha-amylase produced by Bacillus licheniformis. A reliable concentration monitoring experimentation highlighted that the reducing sugars released continuously from YCP during its incubation with the enzyme, and the MW of the main resulting fragment weighed 0.8 x 10(4) Da with approximately 10% of YCP converted to maltose, maltotriose and glucose after a 120-min enzymatic degradation. Finally, YCP was found to be able to increase phagocytic activity of mice in vitro and in vivo, indicating that it may be looked up as a potent immunomodulator that could activate macrophages. PMID- 15885874 TI - Neural plasticity and bilateral movements: A rehabilitation approach for chronic stroke. AB - Stroke interferes with voluntary control of motor actions. Although spontaneous recovery of function can occur, restoration of normal motor function in the hemiplegic upper limb is noted in fewer than 15% of individuals. However, there is increasing evidence to suggest that in addition to injury-related reorganization, motor cortex functions can be altered by individual motor experiences. Such neural plasticity has major implications for the type of rehabilitative training administered post-stroke. This review proposes that noteworthy upper extremity gains toward motor recovery evolve from activity dependent intervention based on theoretical motor control constructs and interlimb coordination principles. Founded on behavioral and neurophysiological mechanisms, bilateral movement training/practice has shown great promise in expediting progress toward chronic stroke recovery in the upper extremity. Planning and executing bilateral movements post-stroke may facilitate cortical neural plasticity by three mechanisms: (a) motor cortex disinhibition that allows increased use of the spared pathways of the damaged hemisphere, (b) increased recruitment of the ipsilateral pathways from the contralesional or contralateral hemisphere to supplement the damaged crossed corticospinal pathways, and (c) upregulation of descending premotorneuron commands onto propriospinal neurons. PMID- 15885875 TI - Blood glucose changes and memory: effects of manipulating emotionality and mental effort. AB - Increasing the emotionality of target material often facilitates memory performance which may be linked to the liberation of glucose. Conversely, increasing mental effort leads to reduced performance and measurable falls in blood glucose. A 2 x 2, parallel groups experiment examined these two phenomena directly by assessing blood glucose levels and memory for neutral and emotional word lists, both with and without a secondary task. Co-performing the secondary task significantly reduced blood glucose in the neutral word condition only and resulted in a global reduction of memory performance in both neutral and emotional word conditions. Processing emotional material resulted in significantly raised blood glucose levels, however, there was no advantage for memory of emotional words. A follow-up study confirmed that the emotionality manipulation was effective. We conclude that there exists a clear relationship between reduced blood glucose and impaired memory performance during periods of mental effort. However, the relationship between physiological and cognitive processes associated with processing emotional material are more complex. PMID- 15885876 TI - Normal N400 in mood disorders. AB - Individuals diagnosed with major depression have been characterized as having a variety of cognitive problems based on a number of behavioral and psychophysiological measures, but it is not clear whether there is a consistent language processing abnormality in depression. Three studies sought to determine whether diverse mood disordered samples show abnormal semantic processing, as indexed by a failure to show increased N400 event-related brain potential amplitudes to passively viewed incongruent, relative to congruent sentence endings. Individuals with major depression (N = 50) or dysthymia (N = 14) had N400 amplitudes similar to those of controls (N = 41) in this sentence processing paradigm. These results are consistent with a small behavioral literature suggesting intact semantic processing in depression and further indicate that abnormal controlled processing in some tasks does not simply reflect a generalized deficit. PMID- 15885877 TI - The effects of antimuscarinic treatments in overactive bladder: a systematic review and meta-analysis. AB - OBJECTIVES: To evaluate the tolerability, safety and efficacy of antimuscarinic drugs used to treat overactive bladder and to identify any differences between individual antimuscarinics. METHODS: Medline, Embase, CCTR and Cinahl databases were searched for published RCTs including an antimuscarinic agent from 1966 to August 2004. Data from included trials were extracted and meta-analysed where possible. RESULTS: Fifty-six trials were included. The antimuscarinics were found to be safe and efficacious. All antimuscarinics apart from oxybutynin IR were found to be well tolerated. Dry mouth was the most commonly reported adverse event and no drug was associated with an increase in any serious adverse event. There were significant differences between the antimuscarinics in rates of withdrawal and rates and range of adverse events and efficacy outcomes. CONCLUSIONS: The antimuscarinics have different tolerability and safety profiles, which are clinically significant. PMID- 15885878 TI - Controlled synthesis of target strings in a class of splicing systems. AB - This article presents an approach for synthesizing target strings in a class of computational models of DNA recombination. The computational models are formalized as splicing systems in the context of formal languages. Given a splicing system (of a restricted type) and a target string to be synthesized, we construct (i) a rule-embedded splicing automaton that recognizes languages containing strings embedded with symbols representing splicing rules, and (ii) an automaton that implicitly recognizes the target string. By manipulating these two automata, we extract all rule sequences that lead to the production of the target string (if that string belongs to the splicing language). An algorithm for synthesizing a certain type of target strings based on such rule sequences is presented. PMID- 15885879 TI - The coactivator Bridge-1 increases transcriptional activation by pancreas duodenum homeobox-1 (PDX-1). AB - Well-orchestrated transcriptional regulation of pancreatic beta cells is essential for insulin production and glucose homeostasis. Pancreas duodenum homeobox-1 (PDX-1) is a key regulator of glucose-dependent insulin production and glucose metabolism. We find that PDX-1 interacts with the PDZ-domain coactivator Bridge-1 in yeast interaction trap assays. Rat Bridge-1 and PDX-1 interact directly in GST pull-down assays via Bridge-1 interactions with the amino terminal transactivation domain of PDX-1. Bridge-1 also interacts with wild-type and mutant human PDX-1 (IPF-1) proteins and strongly interacts with the amino terminal PDX-1 P63fsdelC (MODY4) mutant protein. Transcriptional activation by PDX-1 is increased by addition of Bridge-1 in multiple contexts, including synergistic activation of a Gal4 reporter by Gal4-Bridge-1 and Gal4-PDX-1 fusion proteins, activation of the somatostatin promoter TAAT1 enhancer, and synergistic activation of the rat insulin I promoter FarFlat enhancer by PDX-1, E12, and E47. We propose that the coactivator Bridge-1 modulates PDX-1 functions in the regulation of its target genes. PMID- 15885880 TI - Prolactin signals via Stat5 and Oct-1 to the proximal cyclin D1 promoter. AB - Prolactin (PRL) modulates proliferation in the mammary gland and other tissues, in part through inducing transcription of cyclin D1, a key regulator of G(1) phase cell cycle progression. We showed previously that PRL, via Jak2, induces binding of Stat5 to a distal GAS site (GAS1) in the cyclin D1 promoter. However, full promoter activity requires additional regions, and in this paper we explored PRL-induced activity at sites other than GAS1. We defined a second PRL-responsive region spanning -254 to -180 that contains a second GAS site (GAS2) and an Oct-1 binding site. Although mutational analysis indicated independence from GAS2, proximal promoter activity remained Stat5-dependent, suggesting alternative mechanisms. EMSA showed that Oct-1 binds the -254 to -180 region and that PRL decreased Oct-1 binding, leading to increased PRL-responsiveness of the proximal cyclin D1 promoter in multiple cell lines. This suggests a role for Oct-1 in PRL dependent control of cyclin D1 transcription. PMID- 15885881 TI - Acute reversible parkinsonism in a diabetic-uremic patient. AB - Acute movement disorders with basal ganglia lesions have been recently described in diabetic-uremic patients of Asian descent. The process is often reversible, with a favourable clinical outcome. Metabolic (i.e. uremic toxins) and microangiopathic changes have been suggested to be involved in its pathophysiology, even though racial and/or genetic factors might play a role too. In this report, we present a Caucasian diabetic patient with a long-lasting mild uremia in which acute parkinsonism occurred after a steep and unexpected increase of the serum creatinine. The follow-up demonstrated a significant improvement of the neurological signs and symptoms, the creatinine level lowered close to the premorbid levels, and after several months the patient had fully recovered. Our case history suggests that this unusual clinical syndrome is most probably not restricted to Asian patients. Because its potentially favourable outcome, it should be regularly included in the differential diagnosis of acute movement disorders. PMID- 15885882 TI - DNA methyltransferase expression and DNA hypermethylation in human hepatocellular carcinoma. AB - Aberrant DNA methylation and increased expression of DNA methyltransferases (DNMTs) are features of tumor cells. To investigate roles for DNMTs during hepatocarcinogenesis, we examined DNMT expression at both the mRNA and protein level in hepatocellular carcinomas (HCCs) and paired non-neoplastic liver tissues, along with measuring the DNA methylation status of five tumor suppressor genes. Expression of DNMT1, DNMT3a and DNMT3b mRNA was detected in 33.3, 59.3, and 55.6% of HCCs and 40.7, 22.2, and 0% of non-neoplastic liver tissues, respectively. DNMT1 and DNMT3a were immunoreactive in 100 and 48% of HCCs and 52 and 0% of non-neoplastic liver tissues. The DNMT3a mRNA expression profile showed significant correlation with its immunoreactivity (P=0.022). DNA methylation status of five tumor suppressor genes, HIC-1, p16, RASSF1A, p53, and RB1 was detected in 85.2, 48.1, 44.4, 22.2, and 0% of HCCs, respectively. There was no significant correlation between DNMT mRNA expression and DNA methylation (P>0.05). DNMT immunoreactivity was also not associated with DNA methylation except HIC-1 (P=0.036) and p53 methylation (P=0.009). Despite the lack of correlation between DNA methylation status and DNMT expression, the frequency of hypermethylation of tumor suppressor genes remained relatively high in HCCs, suggesting that regional DNA hypermethylation is involved in hepatocarcinogenesis and that there may be other mechanisms for increasing DNA methylation. PMID- 15885883 TI - Chemopreventive effects of Paullinia cupana Mart var. sorbilis, the guarana, on mouse hepatocarcinogenesis. AB - Guarana (Paullinia cupana) is originally from Amazon, Brazil. Its effects on mouse hepatocarcinogenesis have been investigated in this study. Mice were treated with N-nitrosodiethylamine (DEN), received three different doses of P. cupana added to commercial food, and euthanized after 25 weeks. Gross lesions were quantified, and preneoplastic lesions (PNL) were histologically measured. Cellular proliferation was evaluated by immunobloting for the proliferating cell nuclear antigen (PCNA). The incidence and multiplicity of macroscopic lesions were reduced. The PNL number and PCNA expression were reduced in the highest P. cupana dose. According to these results, guarana presented inhibitory effects on DEN hepatocarcinogenesis in mice. PMID- 15885884 TI - Expression of candidate chromosome 3p21.3 tumor suppressor genes and down regulation of BLU in some esophageal squamous cell carcinomas. AB - The expression of six chromosome 3p21.3 candidate tumor suppressor genes (BLU, FUS2, HYAL2, NPRL2, RASSF1A, and SEMA3B) in esophageal squamous cell carcinoma (ESCC) has been investigated. Reduced expression of BLU was detected in some ESCC cell lines and tumor tissues and the difference was quantitated by real-time quantitative polymerase chain reaction. Methylation specific-PCR revealed the down-regulation of BLU by epigenetic inactivation. However, exogenous expression of BLU did not functionally suppress tumorigenicity in nude mice. These results suggest that over-expression of BLU alone is not sufficient to inhibit tumorigenicity. Further studies on BLU interacting proteins are required to elucidate the possible role of BLU in the development of ESCC. PMID- 15885885 TI - State of hepatitis C viral replication enhances activation of NF-kB- and AP-1 signaling induced by hepatitis B virus X. AB - We examined the effect of hepatitis B virus X (HBx) on NF-kB- and AP-1- mediated transcription in human hepatocellular carcinoma cell lines, Huh-7 with or without subgenomic hepatitis C virus (HCV) RNA. Expression of HBx in Huh-7 cells with HCV resulted in 4.9 times increased NF-kB-activation and 3.8 times AP-1-activation whereas that without HCV resulted in 2.4 times increased NF-kB-activation and 2.3 times AP-1-activation. Interestingly, the expression of the matured form of HCV core protein, Core173, did not activate NF-kB- or AP-1-transcription in either Huh7 with or without HCV replicon. HBx protein might play an important role in HCV-related hepatocarcinogenesis. PMID- 15885886 TI - Transfection of mouse macrophage metalloelastase gene into murine CT-26 colon cancer cells suppresses orthotopic tumor growth, angiogenesis and vascular endothelial growth factor expression. AB - A cDNA fragment coding for domains I and II of mouse macrophage metalloelastase (MME) was transfected into murine CT-26 colon cancer cells that are MME deficient. An orthotopic implantation model was established by using MME transfected cells. In MME-transfected primary tumors, it demonstrated that tumor growth and microvessel formation were significantly inhibited compared with the controls. The expression of vascular endothelial growth factor (VEGF) mRNA and protein was significantly lower in MME-transfected group compared with those in the controls. Our data show that both MME and VEGF gene expression is highly associated with the vascularity of tumors, which may depend on a balance between MME and VEGF expression. PMID- 15885887 TI - Regulation of axin2 expression at the levels of transcription, translation and protein stability in lung and colon cancer. AB - Axin2 is a negative regulator of Wnt/beta-catenin signalling with a role in tumour suppression. Its expression can be up-regulated by E2F1 allowing cross talk between the pRb/E2F and Wnt/beta-catenin pathways, which both have critical roles in the development of many cancers. Thereby, axin2 may have a crucial role in carcinogenesis and here we examine the regulation of its expression. Axin2 mRNAs contain one of three different 5' untranslated regions that can have profound effects on the efficiency of axin2 translation. We show that axin2 mRNA expression is altered in tumours at levels of both total mRNA and relative proportions of alternative 5' untranslated regions. Moreover, the translational efficiencies defined by these 5' untranslated regions are modulated considerably. Additionally, we show that stability of axin2 protein provides a further level of expression regulation. We discuss this complex regulation in terms of axin2's function in carcinogenesis. PMID- 15885888 TI - Potentiation of the activity of cisplatin and cyclophosphamide by trimidox, a novel ribonucleotide reductase inhibitor, in leukemia-bearing mice. AB - We describe the use of the new ribonucleotide reductase inhibitor, trimidox (TDX), in combination chemotherapy under in vitro and in vivo conditions with cisplatin and cyclophosphamide. In vitro, the combination of TDX and cisplatin was tested in L1210 cells. The combination caused concentration dependent antagonistic or additive effects. However, the combination of TDX-cisplatin cyclophosphamide in vivo is highly synergistic in both, the L1210 and P388D1 leukemia mouse models. Both combinations, TDX with cisplatin or TDX with cyclophosphamide were also synergistic in the L1210 and P388D1 leukemia animal models. PMID- 15885889 TI - Exon 3 polymorphisms and haplotypes of O6-methylguanine-DNA methyltransferase and risk of bladder cancer in southern China: a case-control analysis. AB - Methylating agents are involved in bladder carcinogenesis. O6-Methylguanine-DNA methyltransferase (MGMT) is a DNA repair protein that removes methyl group from O6-methylguanine and thus plays an important role in the etiology of cancer. We hypothesized that two MGMT polymorphisms in exon 3, C16195T (or MGMT L53L) and C16286T (or MGMT L84F) are associated with risk of bladder cancer. In a hospital based case-control study of 167 patients with bladder cancer and 204 cancer-free controls frequency-matched by age, sex, smoking status, and alcohol use, we genotyped these two MGMT polymorphisms. We found that these two polymorphisms alone had a non-significant main effect on risk of bladder cancer. However, when these two polymorphisms were evaluated together, individuals with the combined genotypes or haplotypes with one or more variant alleles (i.e. the 16195T and 16286T alleles) had statistically significantly increased risk of bladder cancer (adjusted odd ratio [OR]=1.67, 95% confidence interval [CI], 1.01-2.77) compared with those with no variant allele. In the stratification analysis, the risk of bladder cancer was increased in a dose-response manner as the age increased (P(trend)=0.010), and the increased risk was more pronounced among old subjects (>65 years) (adjusted OR=2.51, 95% CI, 1.05-6.04), men (1.76, 1.00-3.10), and non drinkers (1.91, 1.08-3.36). In conclusion, these two MGMT polymorphisms may jointly play a role, in the etiology of bladder cancer in southern Chinese population. Larger studies are warranted to validate our findings. PMID- 15885890 TI - Conjugated linoleic acid induces apoptosis in MDA-MB-231 breast cancer cells through ERK/MAPK signalling and mitochondrial pathway. AB - We investigated the molecular mechanisms involved in the anti-proliferative activity exerted by conjugated linoleic acid (CLA) on the estrogen unresponsive MDA-MB-231 human breast cancer cell line. The effects on cell proliferation, cell cycle progression and induction of apoptosis were examined. CLA caused the reduction of cell proliferation along with the accumulation of cells in the S phase of the cycle. The occurrence of apoptosis in these cells was indicated by flow cytometry data and further confirmed by the onset of cells with morphological features typical of apoptosis. ERK1/2 reduction and upregulation of pro-apoptotic protein Bak were induced. These events were associated with: (a) reduced levels of the anti-apoptotic protein Bcl-x(L), (b) the translocation of cytochrome c from the mitochondria to the cytosol, (c) the cleavage of pro caspase-9 and pro-caspase-3. From the above data, we are induced to think that CLA may trigger apoptosis in the estrogen unresponsive MDA-MB-231 cell line via mechanisms involving above all the mitochondrial pathway. PMID- 15885891 TI - Plasma carotenoid, alpha-tocopherol and retinol concentrations and risk of colorectal adenomas: A case-control study in Japan. AB - To investigate associations between plasma carotenoids, alpha-tocopherol and retinol with colorectal adenomas risk, we measured concentrations in 224 asymptomatic colorectal adenoma cases and 230 population-based controls matched for age and sex. After adjustment for age, history of colorectal adenomas and cancers, BMI, smoking, drinking status, multivitamin consumption and plasma total cholesterol, the risk of colorectal adenomas in the highest quartile was approximately half of that of men in the lowest quartile for alpha-carotene (OR=0.38; 95% CI: 0.18-0.84; P(trend)=0.01), beta-carotene (OR=0.51; 95% CI: 0.24 1.07; P(trend)=0.03) and total carotenoids (OR=0.48; 95% CI: 0.22-1.03; P(trend)=0.04). In addition, a protective association for alpha-carotene in women was also indicated, but which did not reach statistical significance (OR=0.53; 95% CI: 0.19-1.52; P(trend)=0.35). Our findings suggest a protective effect of carotenoids against the development of colorectal adenomas. PMID- 15885892 TI - Polymorphisms in RAI and in genes of nucleotide and base excision repair are not associated with risk of testicular cancer. AB - Testicular cancer has been suggested to be primed in utero and there is familiar occurrence, particularly brothers and sons of men with testicular cancer have increased risk. Although no specific causative genotoxic agents have been identified, variations in DNA repair capacity could be associated with the risk of testicular cancer. A case-control study of 184 testicular cancer cases and 194 population-based controls living in the Copenhagen Greater Area in Denmark was performed. We found that neither polymorphisms in several DNA repair genes nor alleles of several polymorphisms in the chromosomal of region 19q13.2-3, encompassing the genes ASE, ERCC1, RAI and XPD, were associated with risk of testicular cancer in Danish patients. This is in contrast to other cancers, where we reported strong associations between polymorphisms in ERCC1, ASE and RAI and occurrence of basal cell carcinoma, breast cancer and lung. To our knowledge this is the first study of DNA repair gene polymorphisms and risk of testicular cancer. PMID- 15885893 TI - TWEAK and Fn14: new molecular targets for cancer therapy? AB - Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumor necrosis factor (TNF) superfamily of structurally related cytokines. Full length, membrane-anchored TWEAK can be found on the surface of many cell types and a smaller, biologically active form, generated via proteolytic processing, has also been detected in the extracellular milieu. TWEAK acts via binding to a recently identified TNF receptor superfamily member named fibroblast growth factor-inducible 14 (Fn14). It has been demonstrated that TWEAK binding to the Fn14 receptor, or constitutive Fn14 overexpression, activates the nuclear factor kappaB signaling pathway, which is known to play an important role in immune and inflammatory processes, oncogenesis, and cancer therapy resistance. In this article, we review recent studies indicating that TWEAK and Fn14 may be potential regulators of human tumorigenesis. PMID- 15885894 TI - Differential metastasis-associated gene analysis of prostate carcinoma cells derived from primary tumor and spontaneous lymphatic metastasis in nude mice with orthotopic implantation of PC-3M cells. AB - The purpose of these studies was to explore the genes associated with invasion and metastasis of human prostatic carcinoma line PC-3M in nude mice. After PC-3M cells were inoculated in orthotopic site (prostate) in male nude mice for two months, tumor cells were isolated from primary tumor and lymph node metastasis in the same mouse, respectively. Cell invasion and adhesion ability in vitro were first compared between two cell lines. Then human metastasis-related genes differentially expressed between them were analyzed by utilizing cDNA microarray technique. The in vitro cell invasion and adhesion potential of tumor cells from lymph node metastasis was significantly higher than those from primary tumor, Metastasis-related genes differentially expressed between those two cell lines were identified, all of them were up-regulated in the tumor cells from lymph node metastasis and could be categorized as: (1) genes encoding cellular matrix degrading proteolytic enzyme including cathepsin and MMP; (2) genes encoding transcription factors; (3) genes related to heterotypic adhesion of tumor cells; (4) genes encoding cell surface receptors. Moreover, Four genes were chosen for semi-quantitative RT-PCR analysis, they showed a consistent expression pattern with that of cDNA microarray analysis. We concluded that the lymph node metastasis in nude mice given an injection of PC-3M cells in the prostate is a selective process favoring the survival and growth of a special subpopulation derived from primary tumor with specific genetic alterations, which may play a pivotal role in the metastasis of prostate cancer. Identification and further characterization of these genes may allow a better understanding of lymphatic metastasis in prostate carcinoma. PMID- 15885895 TI - Bioactivation of 3-aminobenzanthrone, a human metabolite of the environmental pollutant 3-nitrobenzanthrone: evidence for DNA adduct formation mediated by cytochrome P450 enzymes and peroxidases. AB - 3-Nitrobenzanthrone (3-NBA) is a suspected human carcinogen found in diesel exhaust and ambient air pollution. The main metabolite of 3-NBA, 3 aminobenzanthrone (3-ABA), was detected in the urine of salt mining workers occupationally exposed to diesel emissions. We evaluated the role of hepatic cytochrome P450 (CYP) enzymes in the activation of 3-ABA in vivo by treating hepatic cytochrome P450 oxidoreductase (POR)-null mice and wild-type littermates intraperitoneally with 0.2 and 2mg/kg body weight of 3-ABA. Hepatic POR-null mice lack POR-mediated CYP enzyme activity in the liver. Using the (32)P-postlabelling method, multiple 3-ABA-derived DNA adducts were observed in liver DNA from wild type mice, qualitatively similar to those formed in incubations using human hepatic microsomes. The adduct pattern was also similar to those formed by the nitroaromatic counterpart 3-NBA and which derive from reductive metabolites of 3 NBA bound to purine bases in DNA. DNA binding by 3-ABA in the livers of the null mice was undetectable at the lower dose and substantially reduced (by up to 80%), relative to wild-type mice, at the higher dose. These data indicate that POR mediated CYP enzyme activities are important for the oxidative activation of 3 ABA in livers, confirming recent results indicating that CYP1A1 and -1A2 are mainly responsible for the metabolic activation of 3-ABA in human hepatic microsomes. No difference in DNA binding was found in kidney and bladder between null and wild-type mice, suggesting that cells in these extrahepatic organs have the metabolic capacity to oxidize 3-ABA to species forming the same 3-ABA-derived DNA adducts, independently from the CYP-mediated oxidation in the liver. We determined that different model peroxidases are able to catalyse DNA adduct formation by 3-ABA in vitro. Horseradish peroxidase (HRP), lactoperoxidase (LPO), myeloperoxidase (MPO), and prostaglandin H synthase (PHS) were all effective in activating 3-ABA in vitro, forming DNA adducts qualitatively similar to those formed in vivo in mice treated with 3-ABA and to those found in DNA reacted with N-hydroxy-3-aminobenzanthrone (N-OH-ABA). Collectively, these results suggest that both CYPs and peroxidases may play an important role in metabolizing 3-ABA to reactive DNA adduct forming species. PMID- 15885896 TI - Fatty acid metabolism pathway play an important role in carcinogenesis of human colorectal cancers by Microarray-Bioinformatics analysis. AB - The present study systematically explored metabolic pathways and altered expressions of genes speculatively participating in colorectal carcinogenesis by using a Microarray-Bioinformatic analysis methods. The results revealed that 157 genes were up-regulated and 281 genes were down-regulated in colorectal cancer (CRC). Gene Ontology (GO) and relevant bioinformatics tools indicated that the functional category to which 438 genes (12%; 438/3800) of the most frequent alteration belonged was metabolism. The analysis of 10 colorectal cancer tissue specimens demonstrated that genes involved in fatty acid metabolic pathways had high rates of overexpression. In addition, we stimulated CRL-1790 cell line with linoleic acid (a polyunsaturated fatty acid) for 12, 24, 48 and 72 h. Cell proliferation was elevated by 5, 25, 28 and 31% (P<0.05), respectively. Further analyses revealed that the genes increasingly expressed in the cell line included enoyl-Coenzyme A, hydratase/3-hydroxyacyl Coenzyme A dehydrogenase (EHHADH), enoyl Coenzyme A hydratase, short chain, 1, mitochondrial (ECHS1); glutaryl Coenzyme A dehydrogenase (GCDH), acyl-Coenzyme A oxidase 2, branched chain (ACOX2); acyl-Coenzyme A dehydrogenase, C-2 to C-3 short chain precursor (ACADS); carnitine palmitoyltransferase 1B (CPT1B), acyl-CoA synthetase long-chain family member 5 (ACSL5), and cytochrome P450, family 4, subfamily A, and polypeptide 11 (CYP4A11) genes. This indicated that the stimulating effect of linoleic acid on cell proliferation was due to interference with the metabolic pathway of fatty acid metabolism. In conclusion, genes with altered expression levels in CRC were mainly associated with fatty acid metabolic pathways speculated to have an important role linked to carcinogenesis. PMID- 15885897 TI - Inhibitory effect of caffeic acid phenethyl ester on the growth of C6 glioma cells in vitro and in vivo. AB - Caffeic acid phenethyl ester (CAPE), a component of honeybee propolis, has been reported to hold various biochemical responses. In the preliminary study, we found that CAPE inhibited the growth of C6 glioma cells in a dose dependent and time dependent manner as shown by the results of trypan blue dye exclusion assay and cell proliferation assay. In addition, the cell number percentage of the G0/G1 phase increased to 85% after the treatment with 50 microM of CAPE for 24h. After treatment with CAPE (50 microM) for 6h, it demonstrated that the protein level of hyperphosphorylated pRb decreased, and cyclin dependent kinase inhibitors p21, p27, and p16 were marked up-regulated. The association of CDK2 and cyclin E that affects the CDK2 activity decreased. When C6 cells were grown as xenografts in nude mice, treatment with CAPE (1-10mg/kg; ip) induced a significant dose dependent decrease in tumor growth by evaluating tumor volume and tumor weight. Histochemical and immunohistochemical analysis revealed that CAPE treatment significantly reduced the number of mitotic cells and proliferating cell nuclear antigen (PCNA)-positive cells in C6 glioma. These results suggest that CAPE presents an antitumor potential for glioma by inhibiting the growth of tumor cells. PMID- 15885898 TI - Safety and security issues relating to low capacity storage of AN-based fertilizers. AB - Motivated by both the Toulouse explosion, and a series of recent unexpected handling and storage accidents in well-developed countries, the safety issues associated with the storage of fertilizer grades of ammonium nitrate (AN) are considered with a focus on low storage capacity premises. Such facilities are numerous and, in large agricultural countries, include thousands of end-users and hundreds of small distributors. The strong oxidative (sometimes explosive) properties of products containing significant amounts of AN have led to a long history of major accidents including mass explosions in large storage units and pre 1950s, to mass explosions in ships. A major breakthrough in safety was achieved in the 1950s, with the promotion - amongst other improvements - of better anti-caking agents. Although modern AN fertilizers complying with current standards are not considered as explosive material per se, the latent risk of accidental detonation under specific conditions remains a real issue, and from a scientific point of view, cannot be completely ruled out-as dramatically demonstrated by the Toulouse disaster in France. The new insight provided here is derived from: (1) a literature review on hazardous properties of AN and AN-based fertilizers; (2) a review of accidents focusing more particularly on the reporting of recent new cases involving relatively small quantities of previously 'thought safe' products; (3) an examination of both the relevant regulatory framework and the level of hazard control achieved; (4) appropriate discussions of the economical, technical and organizational factors that could lead to some underestimation of the risk compared to large scale storage facilities. In terms of research requirements, the complex potential scenario 'mass explosion following a fire' requires further attention, as does the role and properties of molten ammonium nitrate, which could be the precursor for such an event to occur. Beyond research needs, reinforced legislative control by the authorities and further promotion of safe storage practices must be encouraged by the industry for end-users particularly. Such users have inherently the highest potential for undesirable situations, due both to the nature of their activities and also a possible lack of awareness of the real danger. PMID- 15885899 TI - Trunk muscle proprioceptive input assists steering of locomotion. AB - During locomotion, human subjects navigate in their environment and choose the direction by means of the internal representation of space that is continuously updated by sensory input. Aim of this study was to assess whether trunk proprioceptive information plays a role in the definition of the reference frame for orientation. Unilateral trunk muscle vibration was applied during locomotion along a straight path in seven subjects. Vibration was administered either from the onset or in the middle of a seven-step task, under eyes-open (EO) or blindfolded condition. The deviation of the walking trajectory was quantified by the distance of the seventh from the first foot print along the medio-lateral axis. Foot angles and stride lengths were computed for all foot-falls. Vibration produced a clear-cut deviation from the straight-ahead direction when delivered in the middle of blindfolded locomotion. With EO the deviation was much smaller. A mild deviation was obtained in blindfolded condition when vibration started at the onset of locomotion. All deviations from the straight-ahead were accompanied by coherent changes in foot orientation on the ground. Trunk proprioception plays a major role in the definition of locomotor trajectory. Trunk input seems to be weighted against vision and whole-body kinematic information. PMID- 15885900 TI - Endothelin receptors in the detached retina of the pig. AB - Endothelin-1 (ET-1) is a potent vasoconstrictor that causes hypoperfusion of the neurosensory retina. We investigated immunohistochemically the expression of the receptors for ET-1, ET(A) and ET(B), in control and locally detached retinas of the pig. Immunoreactivity for ET(A) was expressed in the innermost retinal layers and in the outer plexiform layer in control retinas, and was additionally strongly expressed by retinal blood vessels at 7 days after detachment of the sensory retina from the pigment epithelium. Immunoreactivity for ET(B) was expressed by the innermost retinal layers, by ganglion cell somata, and by Muller glial cells in the control tissue, and was not altered in its expression after detachment. The vascular expression of ET(A) may suggest a hypoperfusion of the retina after detachment. PMID- 15885901 TI - A role of insulin-like growth factor 1 in beta amyloid-induced disinhibition of hippocampal neurons. AB - In the present study we investigated the effects of beta amyloid (Abeta) on inhibitory synaptic transmission in the cultured hippocampal neurons using whole cell patch-clamp recordings and immunocytochemistry, and examined the role of insulin-like growth factor 1 (IGF-1). Incubation with 4 microM Abeta25-35 for 24 h significantly decreased the frequency of spontaneous inhibitory postsynaptic currents (sIPSCs), but had no effect on the mean amplitude. Pretreatment with 10 ng/ml IGF-1 for 24h prior to Abeta25-35 exposure blocked Abeta-induced disinhibition of hippocampal neurons. The frequency and mean amplitude of miniature IPSC (mIPSCs) were not significantly affected by Abeta. The rise and decay kinetics of sIPSCs and mIPSCs were similar for the control and Abeta25-35 treated hippocampal neurons. Immunocytochemistry showed no changes in the ratio of gamma-aminobutyric acid (GABA) positive cells subsequent to treatment with Abeta, or IGF-1. Together these data suggest that Abeta-induced the disinhibition in cultured hippocampal neurons, whereas IGF-1 could block this effect. PMID- 15885902 TI - A lack of mu-opioid receptors modulates the expressions of neuropeptide Y and substance P mRNA. AB - The present study was undertaken to investigate changes in the expressions of neuropeptide Y (NPY) and substance P (SP) in mice lacking mu-opioid receptors. In an in situ hybridization study, in which we compared wild type and mu-opioid receptor knockout mice, NPY mRNA levels were found to be lower in the caudate putamen and nucleus accumbens of mu-opioid receptor knockout mice. In addition, SP mRNA levels were lower in the ventromedial hypothalamic nucleus of mu-opioid receptor knockout mice. Our findings suggest that a lack of mu-opioid receptors modulates basal NPY mRNA levels in striatal regions and SP mRNA levels in the ventromedial hypothalamic nucleus of the mouse, and that these changes are due to compensatory modulation in the brain. PMID- 15885903 TI - Age-dependent effects of maternal deprivation on oxidative stress in infant rat brain. AB - Developing brain is much more sensitive to all kind of stressors than the developed brain. Early maternal deprivation causes some behavioural and physiological effects on rats. After the birth, there is no endocrinological response to stressors between post-natal 4 and 14th days, which is called stress hyporesponsive period (SHRP) in rats. This hypo-responsiveness is time- and stressor-specific, as some more severe stressors have been shown to induce a stress response. The present study examined the effects of maternal deprivation on oxidative stress in the hippocampus, prefrontal cortex (PFC) and striatum regions of the brain both during and after SHRP of the infant rats. The results showed that maternal deprivation in SHRP increased antioxidant enzyme activities and reduced lipid peroxidation in infant rat brain. However, by the termination of SHRP, maternal deprivation reduced enzyme activities and increased lipid peroxidation. The results indicated that infant brain might be protected in SHRP from maternal deprivation-induced oxidative stress. PMID- 15885904 TI - A multi-joint lower-limb tracking-trajectory test for the assessment of motor coordination. AB - This study aimed to determine whether a lower-limb trajectory-tracking task performed on a leg press machine, that is commonly adopted in both rehabilitation and resistance training settings, could yield reliable assessment of motor coordination in able-bodied individuals. Twenty-two female subjects allocated to two experimental groups were tested and retested after 48-72 h. Group A was fully familiarized with the experimental procedures before each test while group B received only verbal instructions. The unilateral coordination test consisted of target tracking during a simulated half squat including eccentric and concentric actions. In both groups, tracking error showed significant test-retest reliability with ICC values of 0.77-0.80 (p < 0.05). Significant group (A < B) and time (day 2 < day 1) main effects were found for tracking error, while there was no significant influence of action mode and dominance. Tracking error significantly decreased in the group A ( approximately 15%) but not in the group B on retest. Action mode (eccentric versus concentric), side dominance and familiarization on day 1 had no effect on tracking error. However, movement control significantly improved at day 2, thus confirming the occurrence of short term motor learning and the sensitivity of the present trajectory-tracking test. For the first time, a simple test for the assessment of motor coordination during multi-joint closed-kinetic chain action of lower limb muscles has been proposed. Its uniqueness is represented by the specificity for rehabilitation and resistance training settings. Further studies with larger sample groups (e.g., male subjects and patients) and including neurophysiological measurements are needed. PMID- 15885905 TI - Up-regulation of the expression of cocaine and amphetamine-regulated transcript peptide by electroacupuncture in the arcuate nucleus of diet-induced obese rats. AB - It was reported that acupuncture or electro-acupuncture (EA) is effective in reducing the body weight for obese patients, although the mechanisms remain obscure. In a previous study, we have found that rats fed with high-fat (HIF) diet developed diet-induced obesity (DIO) with a concomitant decrease in the hypothalamic content of the cocaine and amphetamine-regulated transcript (CART) peptide, a peptide with anorexiogenic effect. To assess the central effect of EA on DIO rat, we revealed that EA up-regulated the expression of CART peptide in the arcuate nucleus (ARC) of the DIO rats. After feeding with HIF diet for 14 weeks, the DIO rats received EA stimulation three times per week for 4 weeks. The expression of CART peptide in ARC was measured using immunohistochemistry. The plasma ACTH was measured with ELISA. EA caused a reduction of both body weight and energy intake in DIO rats and increased the expression of CART peptide in ARC. The plasma ACTH was increased in response to restraint stress, but EA produced no further increase in ACTH levels. The results suggest that EA can up regulate the expression of CART peptide to approach normal level, resulting in an inhibition of food intake and a reduction of body weight in DIO rats. PMID- 15885906 TI - Effects of an intense training on functional activity of 5-HT(1B) receptors in human peripheral blood lymphocytes. AB - Serotonin (5-HT) is a neurotransmitter and an immune modulator. At the periphery, the serotonergic system appears to possess a regulatory activity via 5-HT 1B receptors. The present study investigated the effects of a 5-day military course following 3 weeks of combat training on the functional activity of 5-HT 1B/1D receptors in peripheral blood lymphocytes of male soldiers. The results of [35S]GTPgammaS binding assays showed that h5-HT 1B/1D receptors were desensitized after the training program, although serum 5-HT was unchanged. These data suggest the existence of a control on T cells mediated through h5-HT 1B/1D receptors leading cytokine production modulation after a physical training. PMID- 15885907 TI - Post-ischemic administration of progesterone in rats exerts neuroprotective effects on the hippocampus. AB - Progesterone is neuroprotective in models of focal or global ischemia when treatment starts either before the insult or at the onset of reperfusion. In these cases the steroid may act during the occurrence of the early pathophysiological events triggered by ischemia or reperfusion. As opposed to this condition, the aim of the present study was to assess the effect of delayed, post-injury administration of progesterone on the preservation of pyramidal neurons of the hippocampus of rats 21 days after been exposed to global ischemia by the four vessel occlusion model. Progesterone (8 mg/kg, i.v.) or its vehicle, were administered at 20 min, 2, 6, and 24h after the end of ischemia. At histological examination, brains of the ischemic vehicle-treated rats showed a severe reduction of the population of pyramidal neurons in the CA1 and CA2 subfields (12% and 29% remaining neurons, respectively), and a less severe neuronal loss in the CA3 and CA4 subfields of the hippocampus (68% and 63% remaining neurons, respectively), as compared to rats exposed to sham procedures. They also showed a two-fold enlargement of the lateral ventricles and 33% shrinkage of the cerebral cortex as compared to the sham group. Progesterone treatment resulted in a significant preservation of pyramidal neurons in CA1 and CA2 (40% and 62% remaining neurons), with no ventricular dilation and only a mild (12%) cortical shrinkage. Results suggest that progesterone is able to interfere with some late pathophysiological mechanisms leading both to selective neuronal damage in the hippocampal CA1 and CA2 subfields, and to shrinkage of the cerebral cortex. PMID- 15885908 TI - Neurophysiological evidence for cortical discrimination impairment of prosody in Asperger syndrome. AB - Asperger syndrome (AS), belonging to the autism spectrum of disorders, is one of the pervasive developmental disorders. Individuals with AS usually have normal development of formal speech but pronounced problems in perceiving and producing speech prosody. The present study addressed the discrimination of speech prosody in AS by recording the mismatch negativity (MMN) and behavioural responses to natural utterances with different emotional connotations. MMN responses were abnormal in the adults with AS in several ways. In these subjects, fewer significantly elicited MMNs, diminished MMN amplitudes, as well as prolonged latencies were found. In addition, the MMN generator loci differed between the subjects with AS and control subjects. These findings were predominant over the right cerebral hemisphere. These results show impaired neurobiological basis for speech-prosody processing at an early, pre-attentive auditory discrimination stage in AS. PMID- 15885909 TI - Valence electron energy-loss spectroscopy in monochromated scanning transmission electron microscopy. AB - With the development of monochromators for (scanning) transmission electron microscopes, valence electron energy-loss spectroscopy (VEELS) is developing into a unique technique to study the band structure and optical properties of nanoscale materials. This article discusses practical aspects of spatially resolved VEELS performed in scanning transmission mode and the alignments necessary to achieve the current optimum performance of approximately 0.15 eV energy resolution with an electron probe size of approximately 1 nm. In particular, a collection of basic concepts concerning the acquisition process, the optimization of the energy resolution, the spatial resolution and the data processing are provided. A brief study of planar defects in a Y(1)Ba(2)Cu(3)O(7 )(delta) high-temperature superconductor illustrates these concepts and shows what kind of information can be accessed by VEELS. PMID- 15885910 TI - Visualization of single-wall carbon nanotube (SWNT) networks in conductive polystyrene nanocomposites by charge contrast imaging. AB - The morphology of conductive nanocomposites consisting of low concentration of single-wall carbon nanotubes (SWNT) and polystyrene (PS) has been studied using atomic force microscopy (AFM), transmission electron microscopy (TEM) and, in particular, scanning electron microscopy (SEM). Application of charge contrast imaging in SEM allows visualization of the overall SWNT dispersion within the polymer matrix as well as the identification of individual or bundled SWNTs at high resolution. The contrast mechanism involved will be discussed. In conductive nanocomposites the SWNTs are homogeneously dispersed within the polymer matrix and form a network. Beside fairly straight SWNTs, strongly bended SWNTs have been observed. However, for samples with SWNT concentrations below the percolation threshold, the common overall charging behavior of an insulating material is observed preventing the detailed morphological investigation of the sample. PMID- 15885911 TI - Mammomonogamus laryngeus (Railliet, 1899) infection in buffaloes in Rio Grande do Sul, Brazil. AB - Parasites of the genus Mammomonogamus affect the respiratory tract of domestic animals. The present study was carried out to determine the presence of Mammomonogamus laryngeus infection and to analyze its lesions in infected buffaloes in Rio Grande do Sul, Brazil, between April and November 1999. The infection rate was 30.5%. In 32 infected buffaloes, the worm pairs collected per animal did not exceed 20. The microscopic diagnosis showed intense polypoid to intramucosal proliferation at the entrance to the pharynx, to which the parasites had adhered, with foci of multifocal hydropic degeneration of the epithelium or individual degeneration of epithelial cells with mild intraepithelial inflammatory infiltrate. The submucosa revealed intense lymphocyte infiltrate extending into the salivary glands. The submucosa also showed formation of structures that resemble lymphoid follicles. PMID- 15885912 TI - Is Wolbachia complicating the pathological effects of Dirofilaria immitis infections? AB - Human and animal parasitic filarial nematodes, which often are the cause of severe disease, harbor intracellular bacteria of the genus Wolbachia (Rickettsiaceae). It is thought that these bacteria play an important role in the pathogenesis and immune response to filarial infection. In order to determine the possible role of Wolbachia in heartworm disease, dogs naturally infected with Dirofilaria immitis were studied for specific antibody response to Wolbachia surface protein (WSP). Antibody subclasses were analyzed to determine immune response polarization. Dogs that died from heartworm disease were necropsied, and various organs were studied by immunohistochemistry to determine whether Wolbachia-derived molecules were present in tissue from infected dogs. Humoral response to the WSP was present in all infected dogs and appeared to be predominantly of the Th1-type. Several organs, including lung, liver, and kidney, contained positive-staining cells for WSP, confirming that the canine host does come into contact with Wolbachia-derived molecules. PMID- 15885913 TI - Is heartworm disease really spreading in Europe? AB - Based on recently published surveys and newly acquired data, a study was conducted to verify the distribution of filarial worm (Filarioidea) infections in Europe, with particular emphasis on canine heartworm infection (Dirofilaria immitis). A Geographic Information System based on thermal regimen was constructed as a means to identify areas potentially suitable for heartworm transmission, taking into account that the development of D. immitis larvae in the mosquito does not occur below the threshold temperature of approximately 14 degrees C. Furthermore, a bionomic model of D. immitis in its mosquito vectors, which calculates the moving cumulative heartworm development unit parameter, was applied using the available temperature data to assess the theoretic transmission timing of heartworm in Europe. The results show that the earliest infection risk occurs in Spain on March 21 and the latest risk occurs in Spain on September 11. The longest risk period occurs in Spain (Murcia station: March 21-November 11), and the shortest risk period occurs in northeastern Europe. The study also provides the first risk assessment maps for Europe and suggests that if the actual climatic trend continues, filarial infection should spread into previously infection-free areas. PMID- 15885914 TI - The incidence of calf morbidity and mortality due to vector-borne infections in smallholder dairy farms in Kwale District, Kenya. AB - An observational longitudinal study was carried out on 92 randomly selected smallholder farms in two coastal lowland zones of Kwale District in Kenya between December 1997 and November 1999. The objective was to estimate the incidence of the main vector-transmitted diseases in pre-weaned calves. From an initial 41 pure or cross-bred Bos taurus calves which were less than 2 months and whose birth and disease histories were known, study calves were recruited progressively and monitored until they were weaned at around 146 days. Overall, 130 calves in 67 farms were monitored and these contributed a total risk period of 30,062 days. Disease parameters were analysed and compared as true annual and age-specific incidence rates. The incidences of East Coast fever (ECF) (23.1%) and trypanosomosis (29.1%) were the highest among the vector-borne diseases. The corresponding mortality incidence rates of ECF and trypanosomosis were 10.9 and 3.6%, respectively. The annual incidence rates of anaplasmosis and babesiosis were 10.9 and 1.2%, respectively. There was no mortality arising specifically from anaplasmosis or babesiosis. Analysis of survival times to natural infection indicated that the field challenge resulting to cases of trypanosomosis was much higher compared to the risk of either ECF or anaplasmosis. It was concluded that these vector-borne diseases constrain production of replacement stock in this coastal lowlands region of Kenya. PMID- 15885915 TI - Neurochemical brain groups activated after an isotonic blood volume expansion in rats. AB - In order to establish the involvement of particular neurochemical brain groups in the response to blood volume expansion, we analyzed Fos-labeling in combination with immunolabeling for serotonin, tyrosine hydroxylase, vasopressin and oxytocin, 90 min after a sham or i.v. isotonic blood volume expansion (BVE) in unanesthetized, unrestrained rats. We also examined the changes in concentration of oxytocin, atrial natriuretic peptide and vasopressin plasma, induced by blood volume load, to confirm our previous studies. The results demonstrate the participation of specific paraventricular and supraoptic nucleus groups of cells (oxytocinergic-vasopressinergic), serotoninergic dorsal raphe nucleus cells and catecholaminergic A1/A2/A6 groups (in the caudal ventrolateral medulla, nucleus of the solitary tract and locus coeruleus respectively), in the regulatory response to BVE. They provide detailed neuroanatomical evidence to support previous observations showing the contribution of these neurochemical systems in the neural, behavioral and endocrine response to isotonic BVE. PMID- 15885916 TI - Role of galanin receptor 1 and galanin receptor 2 activation in synaptic plasticity associated with 3',5'-cyclic AMP response element-binding protein phosphorylation in the dentate gyrus: studies with a galanin receptor 2 agonist and galanin receptor 1 knockout mice. AB - The neuropeptide galanin was shown to impair cognitive performance and reduce hippocampal CA1 long-term potentiation (LTP) in rodents. However, the contribution of the two main galanin receptors; GalR1 and GalR2, present in the hippocampus to these effects is not known. In the present study, we determined the protein expression levels of GalR1 and GalR2 in the mouse dentate gyrus (DG) and used galanin (2-11), a recently introduced GalR2 agonist, and GalR1 knockout mice to examine the contribution of GalR1 and GalR2 to the modulation of LTP and 3',5'-cyclic AMP response element-binding protein (CREB)-dependent signaling cascades. In the DG, 57+/-5% of the galanin binding sites were GalR2, and the remaining population corresponded to GalR1. In hippocampal slices, galanin (2-11) fully blocked the induction of DG LTP, whereas galanin (1-29), a high affinity agonist for both GalR1 and GalR2, strongly but not fully attenuated the late phase of LTP by 80+/-1.5%. Application of galanin (1-29) or galanin (2-11) after LTP induction caused a transient reduction in the maintenance phase of LTP, with the larger effect displayed by superfusion of galanin (2-11). The induction and maintenance of DG LTP was not altered in the GalR1 knockout mice. Superfusion of galanin (1-29) or galanin (2-11) blocked the LTP induction to the same degree indicating a role for GalR2 in the induction phase of DG LTP. Furthermore, we analyzed the effects of GalR1 and/or GalR2 activation on DG LTP-induced CREB phosphorylation, associated with the late transcriptional effects of LTP. In the lateral part of the granule cell layer, high-frequency trains stimulation caused a significant increase in the level of CREB phosphorylation, which was significantly reduced by application of either galanin (1-29) or galanin (2-11), indicating that both GalR1 and/or GalR2 can mediate some of their effects on LTP through inhibition of CREB-related signaling cascades. PMID- 15885917 TI - Phosphorylated extracellular signal-regulated kinase 1/2 immunoreactivity identifies a novel subpopulation of sympathetic preganglionic neurons. AB - Distinct chemical codes are thought to reflect functional specificity in sympathetic preganglionic neurons (SPN). Although a number of chemical candidates have been identified including neurotransmitter-related, calcium-binding and other proteins, signal transduction proteins have been largely neglected. Not only might these chemicals allow discrimination of functionally unique chemical signatures, but they may also identify activated neurons. Immunoreactivity (ir) to phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) was differentially located within the thoracic spinal cord depending upon which of three forms of killing was used: the only exception to this was the intermediolateral cell column (IML) which was consistently, densely labeled. The presence or absence of p-ERK1/2 in SPN (n=17,541) within the IML of the thoraco lumbar spinal cord was determined in seven rats. SPN were identified on the basis of their location, size and that they contained choline acetyltransferase ir. On average, 58% of SPN contained p-ERK1/2, however, more SPN in both the upper (72%; C8-T4) and lower (78%; T11-L3) thoraco-lumbar spinal cord contained p-ERK1/2-ir than the middle thoracic region (47%; T4-T10). p-ERK1/2-ir was also examined in SPN (n=1895) innervating the adrenal medulla (identified by retrograde tracing using cholera toxin B subunit) combined with localization of neuronal nitric oxide synthase (nNOS) in three rats. On average, 64% of adrenal SPN contain p ERK1/2-ir, and it was confirmed that all adrenal SPN contain nNOS-ir. It appears that p-ERK1/2-ir SPN, described in this study, have tonically activated receptors that are coupled to intracellular signal transduction pathways that lead to the phosphorylation of ERK1/2. PMID- 15885918 TI - Tetanus-induced re-activation of evoked spiking in the post-ischemic dentate gyrus. AB - This study aimed at investigating and influencing the basic electrophysiological functions and neuronal plasticity in the dentate gyrus in freely moving rats at several time-points after global ischemia. Although neuronal death was induced selectively in the cornu ammonis, subfield 1 (CA1)-region of the hippocampus, we found an additional loss of the population spike in the dentate gyrus after stimulation of the perforant path. Input/output-measurements revealed that as early as 1 day post-ischemia population spike generation in the granular cell layer is greatly decreased when compared with pre-ischemic values and to sham operated animals, despite an apparently intact morphology of granular cells as evidenced by Nissl-staining. In contrast, the synaptic transmission (excitatory postsynaptic field potential) shows no significant difference when comparing values before and after ischemia and ischemic and sham-operated animals. Despite reduced output function, indicated by very small population spike amplitudes, long lasting potentiation can be induced 10 days after ischemia. Surprisingly, even "silent" populations of neurons, which appear selectively post-ischemia and do not show any evoked population spike, can be re-activated by tetanisation which is followed by a normal appearing long-term potentiation. However, this functional recovery seems to be partial and transient under current conditions: population spike-values do not reach pre-ischemic values and return to the low pre-tetanic baseline values the next day. Electrophysiological measurements ex vivo after ischemia indicate that the neuronal dysfunction in the dentate gyrus is not due to locally destroyed structures but that the activity of granular cells is merely suppressed only under in vivo conditions. In summary, global ischemia leaves a neighboring morphologically intact input area, functionally impaired. However, neuronal function can be partially regenerated by electrophysiological tetanic stimulation. PMID- 15885919 TI - Mapping of brain networks involved in consolidation of lamb recognition memory. AB - In sheep, ewes at parturition are responsive to any newborn lamb, but within less than 1 h, mothers learn to recognize the odor of their lamb and restrict maternal care to their own offspring (maternal selectivity). In a first experiment, we investigated the long-term retention of maternal selectivity after various mother young contact and separation durations. After 4 h of contact, 36 h of separation leads to a total loss of selectivity. Increasing contact duration to 7 days prior to this separation maintains selectivity. These data suggest that lamb memory after going through an initial labile state after parturition, is consolidated over time into a more stable long-term memory. Fos immunohistochemistry reveals that reintroduction of the lamb after 4 h of mother-young contact and 3 h of separation activates different maternal brain regions than reintroduction of the lamb after 7 days of mother-young contact and 3 h of separation. While the piriform cortex shows an enhanced activation at both times, a selective enhancement of activation is observed in the frontal medial and orbitofrontal cortices only after 7 days of mother-young contact. These data suggest that as consolidation occurs, the neurobiological networks sustaining lamb memory involve different structures. PMID- 15885920 TI - Glial fibrillary acidic protein mRNA levels in the cingulate cortex of individuals with depression, bipolar disorder and schizophrenia. AB - Recent studies have shown a decrease in glial number and glial fibrillary acidic protein (GFAP) levels in the frontal and cingulate cortices of individuals with mood disorders and schizophrenia. In an attempt to verify and expand these findings we examined GFAP messenger ribonucleic acid (mRNA) levels in postmortem sections of the anterior cingulate cortex (ACC) from the Stanley Neuropathology Consortium (SNC). The consortium consists of 15 cases in each of four groups (schizophrenia, bipolar disorder, non-psychotic depression and unaffected controls). By in situ hybridization, we found higher levels of GFAP mRNA in white matter and at the pial surface as compared with gray matter levels in all cases. In the white matter of ACC we detected a significant effect of diagnosis (P<0.04) with GFAP mRNA levels decreased in individuals with schizophrenia and bipolar disorder as compared with normal controls. In the gray matter there was a significant effect of layer (P<0.01) with the highest levels of GFAP mRNA in layer VI in all groups. As in the white matter, the mean GFAP mRNA levels were decreased in individuals with schizophrenia and bipolar disorder as compared with the unaffected controls, however the difference failed to reach statistical significance. Thus, astrocytes positive for GFAP may contribute to the decrease in glial density previously described in subjects with major mental illness, however the relative contribution of astrocytes may vary with diagnosis. PMID- 15885921 TI - Ectopic galanin expression and normal galanin receptor 2 and galanin receptor 3 mRNA levels in the forebrain of galanin transgenic mice. AB - The functional interactions of the neuropeptide galanin (GAL) occur through its binding to three G protein-coupled receptor subtypes: galanin receptor (GALR) 1, GALR2 and GALR3. Previously, we demonstrated that GALR1 mRNA expression was increased in the CA1 region of the hippocampus and discrete hypothalamic nuclei in galanin transgenic (GAL-tg) mice. This observation suggested a compensatory adjustment in cognate receptors in the face of chronic GAL exposure. To evaluate the molecular alterations to GALR2 and GALR3 in the forebrain of GAL overexpressing mice, we performed complementary quantitative, real-time PCR (qPCR), in situ hybridization, and immunohistochemistry in select forebrain regions of GAL-tg mice to characterize the neuronal distribution and magnitude of GAL mRNA and peptide expression and the consequences of genetically manipulating the neuropeptide GAL on the expression of GALR2 and GALR3 receptors. We found that GAL-tg mice displayed dramatic increases in GAL mRNA and peptide in the frontal cortex, posterior cortex, hippocampus, septal diagonal band complex, amygdala, piriform cortex, and olfactory bulb. Moreover, there was evidence for ectopic neuronal GAL expression in forebrain limbic regions that mediate cognitive and affective behaviors, including the piriform and entorhinal cortex and amygdala. Interestingly, regional qPCR analysis failed to reveal any changes in GALR2 or GALR3 expression in the GAL-tg mice, suggesting that, contrary to GALR1, these receptor genes are not under ligand-mediated regulatory control. The GAL-tg mouse model may provide a useful tool for the investigation of GAL ligand receptor relationships and their role in normal cognitive and affective functions as well as in the onset of neurological disease. PMID- 15885922 TI - Spinal phospholipase A2 in inflammatory hyperalgesia: role of the small, secretory phospholipase A2. AB - Current work emphasizes that peripheral tissue injury and inflammation results in a heightened sensitivity to subsequent noxious input (hyperalgesia) that is mediated in large part by the spinal synthesis and release of eicosanoids, in particular prostaglandins. Secreted phospholipase A(2)s (sPLA(2)s) form a class of structurally related enzymes that release arachidonic acid from cell membranes that is further processed to produce eicosanoids. We hypothesized that spinal sPLA(2)s may contribute to inflammation-induced hyperalgesia. Spinal cord tissue and cerebrospinal fluid were collected from rats for assessment of sPLA(2) protein expression and sPLA(2) activity. A basal sPLA(2) protein expression and activity was detected in spinal cord homogenate (87+/-17 pmol/min/mg), though no activity could be detected in cisternal cerebrospinal fluid, of naive rats. The sPLA(2) activity did not change in spinal cord tissue or cerebrospinal fluid assessed over 8 h after injection of carrageenan into the hind paw. However, the sPLA(2) activity observed in spinal cord homogenates was suppressed by addition of LY311727, a selective sPLA(2) inhibitor. To determine the role of this spinal sPLA(2) in hyperalgesia, we assessed the effects of lumbar intrathecal (IT) administration of LY311727 in rats with chronic IT catheters in three experimental models of hyperalgesia. IT LY311727 (3-30 microg) dose-dependently prevented intraplantar carrageenan-induced thermal hyperalgesia and formalin induced flinching, at doses that had no effect on motor function. IT LY311727 also suppressed thermal hyperalgesia induced by IT injection of substance P (30 nmol). Using in vivo spinal microdialysis, we found that IT injection of LY311727 attenuated prostaglandin E(2) release into spinal dialysate otherwise evoked by the IT injection of substance P. Taken together, this work points to a role for constitutive sPLA(2)s in spinal nociceptive processing. PMID- 15885923 TI - Binge drinking in the context of romantic relationships. AB - This study examined consequences of binge drinking on relationship conversation qualities (positive tone, general disagreements, drinking disagreements, and talks about drinking) among romantically involved college students. Conversation qualities were predicted with three binging variables: (a) same day binging, (b) prior day binging, and (c) total amount of binging reported. The participants (N=156) completed 10 daily diaries of relationship conversations as well as drinking behaviors. Same day binging increased the occurrence of both drinking disagreements and talks about drinking. However, prior day binging was not associated with any of the four conversation qualities examined. Greater numbers of binges over the duration of the study were associated with less overall positive tone, and more general disagreements, drinking disagreements, and talks about drinking. The implications of the findings for student drinking patterns and relational quality are discussed. PMID- 15885924 TI - Chronic ACTH autoantibodies are a significant pathological factor in the disruption of the hypothalamic-pituitary-adrenal axis in chronic fatigue syndrome, anorexia nervosa and major depression. AB - Dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis is a commonly recognized feature of many pathological conditions. Abnormal adrenal responses to experimental manipulation have been well documented in patients suffering from chronic fatigue syndrome, anorexia nervosa and major depression. Yet no defect of any single organ, gland or brain region has been identified as a cause of these abnormalities. The disruption of the HPA axis that occurs in these conditions can be understood if an interfering factor is present in these patients. Evidence indicates that this interfering factor is adrenocorticotropin hormone (ACTH) autoantibodies. Chronic high levels of ACTH autoantibodies will significantly disrupt the HPA axis and force the body to compensate for an impaired cortisol response. The resulting effect of chronic ACTH autoantibody interference is the manifestation of adrenocortical insufficient symptoms and psychological disturbances. Some symptoms of chronic fatigue syndrome, anorexia nervosa and major depression, such as anxiety, are the adverse effects of mechanisms compensating for less effective ACTH due to autoantibodies. Furthermore, these patients engage in extraordinary behaviors, such as self-injury, to increase their cortisol levels. When this compensation is inadequate, symptoms of adrenocortical insufficiency appear. Corticosteroid supplements have been demonstrated to be an effective treatment for chronic fatigue syndrome, anorexia nervosa and major depression. It allows the patients to have the corticosteroids they require for daily functioning and daily stressors. This therapy will relieve the patients of their symptoms of adrenocortical insufficiency and permit their cortisol-stimulating mechanisms to operate at levels that will not cause pathological problems. PMID- 15885925 TI - Production of glucosinolate hydrolysis products in Farsetia aegyptia suspension cultures following elicitation. AB - Levels of glucosinolates in Farsetia aegyptia var. ovalis suspension cultures were monitored after treatment with yeast extract, chitosan, methyl jasmonate, ampicillin, and Phytophthora infestans autoclaved mycelia as elicitors. Glucosinolates were identified, and an estimation of their levels obtained from their hydrolysis products. Yeast extract improved glucotropaeolin (benzyl glucosinolate) and glucocheirolin [3-(methylsulfonyl)propyl-glucosinolate] accumulation, and production of sec-butyl, isobutylglucosinolate and gluconasturtiin (2-phenylethyl-glucosinolate) was only detected in yeast elicited cultures. Increases were shown in glucotropaeolin levels in cultures elicited using methyljasmonate, and chitosan and methyljasmonate in combination. PMID- 15885926 TI - Antiinflammatory activity of the pectic polysaccharide from Comarum palustre. AB - A pectic polysaccharide named comaruman (CP) was extracted from the aerial part of Comarum palustre with 0.7% aqueous ammonium oxalate and subsequent precipitation with ethanol. Oral administration of comaruman (5-100 mg/kg) was found to reduce a paw edema observed 24 h after injection of 2% formalin in mice. A fraction of comaruman (CP-H9) exhibited a similar antiinflammatory activity. Comaruman, CP deprived of lipid, CP purified by proteins and CP fractions obtained with acidic hydrolysis inhibit spontaneous and phorbol-12-myristate-13 acetate-activated adhesion of peritoneal leukocytes in vitro. PMID- 15885927 TI - Antidiabetic and antihyperlipemic effects of Clemeo felina. AB - Petroleum ether and benzene extracts of Clemeo felina, given orally at doses of 300 mg kg(-1) day(-1) for 30 days, were found to be antidiabetic and antihyperlipemic on alloxan diabetic rats. Moreover, a significant decrease in the activities of serum enzymes like alkaline phosphatase, acid phosphatase and HMGCoA reductase activity in the liver was observed. However, treatment of rats with the extracts as well as standard antidiabetic drugs increased liver hexakinase activity and serum LDH activity. PMID- 15885928 TI - Prevalences and transmission routes of Campylobacter spp. strains within multiple pig farms. AB - In this work, faecal samples were collected from 15 pig farms to determine the Campylobacter prevalences at different times during the rearing period and to visualize the exchange of strains among the pig population by genotyping specific isolates. All isolated strains were identified as C. coli. Whereas no Campylobacter were detectable in the faeces of piglets at the day of birth, the Campylobacter incidence rose within days to 32.8%. After transfer to the nursery unit the prevalence increased to 56.6%. Approximately two-thirds of the pigs remained C. coli shedders in the fattening unit. In contrast to most farms, one farm expressed a very low Campylobacter incidence during the whole rearing period. Amplified fragment length polymorphism (AFLP) analysis was performed on all C. coli isolates of one farm. Clonal strains were identified from the brood sows and their offsprings or neighbouring piglets. After moving to the nursery unit, new genotypes appeared in that pig group but the original C. coli strains largely remained within that group. C. coli genotypes, identified during the fattening period, replaced the previously isolated genotypes. Transportation to the abattoir had no significant influence on the shedding rate of C. coli. The detection rate before transportation was 79.1% and decreased slightly to 78.2% (n=330). Additionally, eleven of 1474 environmental samples from different sources of the pig farms were positive for C. coli. This study demonstrates the importance of pigs as a reservoir for C. coli. Maternal C. coli strains are the primary source of infection but non-related genotypes from different sources appear during the rearing period and these latter strains constitute largely the final C. coli flora. PMID- 15885929 TI - Genetic immunization with codon-optimized equine infectious anemia virus (EIAV) surface unit (SU) envelope protein gene sequences stimulates immune responses in ponies. AB - In the context of DNA vaccines the native equine infectious anemia virus (EIAV) envelope gene has proven to be an extremely weak immunogen in horses probably because the RNA transcripts are poorly expressed owing to an unusual codon-usage bias, the possession of multiple RNA splice sites and potential adenosine-rich RNA instability elements. To overcome these problems a synthetic version of sequences encoding the EIAV surface unit (SU) envelope glycoprotein was produced (SYNSU) in which the codon-usage bias was modified to conform to that of highly expressed horse and human genes. In transfected COS-1 cell cultures, the steady state expression levels of SYNSU were at least 30-fold greater than equivalent native SU sequences. More importantly, EIAV-specific humoral and lymphocyte proliferation responses were induced in ponies immunized with a mammalian expression vector encoding SYNSU. However, these immunological responses were unable to confer protection against infection with a virulent EIAV strain. PMID- 15885930 TI - Molecular characterization of Helicobacter pylori strains isolated from cynomolgus monkeys (M. fascicularis). AB - We recently reported the occurrence of natural infection with H. pylori in a group of cynomolgus monkeys with chronic active gastritis and gastric erosions. The goal of the present study was to characterize and to compare strains isolated from animals originating from two different geographical areas. Gross and microscopic pathology determined at the time of necropsy was similar in all animals. H. pylori were isolated from specimens harvested in five monkeys (four from Vietnam and one from the Philippines) with gastritis. Isolates from monkeys bred in Vietnam had a similar DNA fingerprint pattern, which was distinct from that of isolates from a monkey bred in the Philippines. All strains were of the s1a vacA subtype, but all the 'Vietnamese' strains were cagA+ and all but one were iceA1 whereas the 'Philippino' strains were cagA- and iceA2. The sequences of the 16S rRNA of the Vietnamese and Philippino strains shared 98% homology and both clustered with H. pylori sequences present in the NCBI database. In conclusion, cynomolgus monkeys can be naturally colonized by H. pylori, and the strains isolated from these animals appear to vary according to the geographical origin, thus indicating probable infection prior to importation. Since some of the cynomolgus monkeys developed antral erosions during natural infection, we propose that this animal model may be used to investigate the role of H. pylori in ulcerogenesis. PMID- 15885931 TI - Detection of Mycobacterium avium subsp. paratuberculosis in intestinal and lymph node tissues of water buffaloes (Bubalus bubalis) by PCR and bacterial culture. AB - The efficacy of bacterial culture and IS900-specific polymerase chain reaction (PCR) was compared for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) from the intestinal and mesenteric lymph node tissues of water buffaloes (Bubalus bubalis) showing lesions of paratuberculosis (Johne's disease). Out of 20 (4.9%) animals showing histological lesions suggestive of paratuberculosis, 14 (70%) and 6 (30%) were positive in the PCR and bacterial culture, respectively. The results of this study suggested that PCR was more sensitive than bacterial culture in detection of subclinical paratuberculosis in water buffaloes. The bacterial concentration from large amount of tissues by differential and density gradient centrifugation method was found to facilitate the diagnosis by smear examination and PCR. The specificity of the PCR was confirmed by the product size and restriction digestion pattern of the amplicons. The sequence analysis of the amplified products (626bp of IS900 gene) from buffalo strain showed more than 97% homology with the published sequences. PMID- 15885932 TI - Salmonella entry: M cells versus absorptive enterocytes. PMID- 15885933 TI - Molecular analysis of a 444 bp fragment of the bovine leukaemia virus gp51 env gene reveals a high frequency of non-silent point mutations and suggests the presence of two subgroups of BLV in Chile. AB - With the aim of achieve a better understanding of the epidemiology and distribution of bovine leukaemia virus (BLV) infection in Chile, we assessed the suitability of using DNA isolated from the leukocyte fraction of bulk milk samples to carry out PCR-RFLP and DNA sequence analysis. The env fragment of BLV was successfully amplified from 33 serologically positive bulk milk samples collected from different geographical areas in the south of Chile. Restriction analysis allowed to classify 17 isolates within the Australian subgroup and 16 within the Belgium subgroup. DNA sequence and multiple alignment analysis of eight Chilean isolates showed a significantly higher frequency of single and double nucleotide substitutions. Most of these mutations were non-silent, resulting in changes at the protein level in several important epitopes of gp51. The Chilean sequences and 59 BLV env sequences available at GenBank, were subjected to a phylogenetic analysis, resulting in four different clusters. The groups identified were not related to those previously defined by restriction analysis. Chilean isolates were included in two different clusters and were genetically not related to isolates collected from neighbouring countries. Considering our results we can conclude: (i) bulk milk samples are suitable to identify the presence of BLV allowing epidemiological and genetic studies to be conducted on large geographical areas; (ii) at least four different genetic groups of BLV were identified by phylogenetic analysis, with Chilean isolates included in two different sub clusters. PMID- 15885934 TI - Pregnancy effects on distribution of progesterone receptors, oestrogen receptor alpha, glucocorticoid receptors, Ki-67 antigen and apoptosis in the bovine interplacentomal uterine wall and foetal membranes. AB - Until recently, studies dealing with the uterus of the pregnant cow focus primarily on the placentome or on early and late pregnancy. Thus, there is a paucity of information about many aspects of the interplacentomal uterine wall including adherent foetal membranes. Corresponding tissue specimens were collected at the slaughterhouse and in animals undergoing premature caesarean section. Two specimens per month of pregnancy were assessed immunohistochemically for progesterone receptors, oestrogen receptor alpha and glucocorticoid receptors, Ki-67 protein and TUNEL procedure was performed. The latter two methods were employed in three animals each per months 1 and 2, 3 and 4, 7 and 8 and in six animals undergoing caesarean section at days 274 and 275 post insemination or during spontaneous labour. Results indicate that proliferation and apoptosis are of minor importance for tissue homeostasis since both can histochemically be detected only sporadically. Thus, at the sites investigated here, cellular hypertrophy plays an important role for tissue growth during pregnancy. Progesterone receptors, oestrogen receptor alpha and glucocorticoid receptors, however, exhibit cell type and pregnancy stage specific distribution patterns within the tissues assessed. Progesterone receptor immunoreactive scores remained fairly unchanged during pregnancy. Oestrogen receptor alpha scores, however, generally decreased and glucocorticoid receptors increased with ongoing gestation. Progesterone receptors and oestrogen receptor alpha were present in endometrial stroma and in myometrial smooth muscle cells during whole pregnancy. Oestrogen receptor alpha was detectable during whole pregnancy also in uterine glands. Progesterone receptors were, however, present at a very low level at the latter site only during months 1-3 and 6-9. Oestrogen receptor alpha and glucocorticoid receptors may also mediate uterine blood flow since they were present in the tunica media of uterine blood vessels. Results of the present study indicate, that progesterone and its receptor play an important role during whole gestation, mainly for uterine quiescence. Glucocorticoids and their receptors - possibly in cooperation with oestrogens and decreasing amounts of the oestrogen receptor alpha - should trigger processes initiating parturition, such as endometrial prostaglandin production. Further studies - including the periparturient period - should help to understand the exact role of the extraplacental compartment of the uterine wall for the initiation and progress of parturition. PMID- 15885935 TI - Cyclodextrin solubilization of the antibacterial agents triclosan and triclocarban: formation of aggregates and higher-order complexes. AB - It is well known that water-soluble cyclodextrins form inclusion complexes with many lipophilic water-insoluble drugs and that such complexation frequently enhances the aqueous solubility of drugs. It is also well known that various excipients, such as water-soluble polymers, organic acids and bases and metal ions can enhance the solubilizing effects of cyclodextrins. However, it is not clear how these excipients enhance the effects. The effects of cyclodextrins, 2 hydroxypropyl-beta-cyclodextrin (HPbetaCD) and randomly methylated beta cyclodextrin (RMbetaCD) on the aqueous solubility of triclosan and triclocarban were investigated. The phase-solubility profiles were all of type A(P) indicating formation of higher-order complexes or complex aggregates. Addition of lysine and other excipients enhanced the RMbetaCD solubilization of triclocarban. NMR spectroscopic studies, including 2D ROESY and 1D gROESY techniques, indicated that HPbetaCD and RMbetaCD, as well as their complexes, form aggregates of two to three cyclodextrin molecules. The critical concentration for the aggregate formation was determined to be 5.4% (w/v). Lysine, polyvinylpyrrolidone and magnesium ions formed non-inclusion complexes resulting in formation of multiple component cyclodextrin complexes in aqueous solutions with triclocarban. PMID- 15885936 TI - Asymmetric membrane capsules for delivery of poorly water-soluble drugs by osmotic effects. AB - A non-disintegrating polymeric capsule system, in which asymmetric membrane offers an improved osmotic effect, was used to deliver poorly water-soluble drugs in a control manner. The capsule wall membrane was made by a phase inversion process, in which asymmetric membrane was formed on stainless-steel mold pins by dipping the mold pins into a coating solution containing a polymeric material followed by dipping into a quench solution. This study evaluates the influence of coating formulation that was cellulose acetate (CA), ethylcellulose (EC), and plasticizer (glycerin and triethyl citrate). Results show capsule that made by CA with glycerin (formulation A), which appear in asymmetric structure and are able to release chlorpheniramine maleate (CM) in significant percentage. Two poorly water-soluble drugs of felodipine (FL) and nifedipine (NF) were selected as the model drug to demonstrate how the controlled release characteristics can be manipulated by the design of polymeric capsules with an asymmetric membrane and core formulations. Results show that sodium lauryl sulfate (SLS) is able to promote the release of FL from polymeric capsules prepared with CA with asymmetrical membrane. The addition of solubilizer, including RH40, PVP K-17, and PEG 4000 could enhance the release of FL but with an extent not being related to its solubility. Based on these results, influence of core formulation variables, including the viscosity and added amount of hydroxypropyl methylcellulose (HPMC), the added amount of SLS, and drug loading were examined on the release of NF. It was found that HPMC of 50 cps was suitable to be a thickening agent and both added amount of HPMC and SLS showed a comparable and profoundly positive effect, whereas NF loading had no influence on the drug release percent and rate. There existed a synergistic interaction between HPMC and SLS on the release percent and rate. PMID- 15885937 TI - The effect of packaging materials on the stability of sunscreen emulsions. AB - The purpose of this research was to study the stability of a emulsion containing UVA, UVB and infrared sunscreens after storage in different types of packaging materials (glass and plastic flasks; plastic and metallic tubes). The samples, emulsions containing benzophenone-3 (B-3), octyl methoxycinnamate (OM) and Phycocorail, were stored at 10, 25, 35 and 45 degrees C and representative samples were analyzed after 2, 7, 30, 60 and 90 days period. The stability studies were conducted by analyzing samples at pre-determined intervals by high performance liquid chromatography (HPLC) along with periodic rheological measurements. PMID- 15885938 TI - Evaluation of Eudragit-coated chitosan microparticles as an oral immune delivery system. AB - Chitosan microparticles containing ovalbumin (OVA), OVA-containing chitosan microparticles (Chi-OVA), were prepared, coated with Eudragit L100 (ER), and evaluated as oral vaccine. Chi-OVA with an OVA content of 34.4% (w/w) and a mean particle size of 2.3 microm were used for experiments in vitro and in vivo. ER coated Chi-OVA (ER-Chi-OVA) contained 3.6-20.5% (w/w) OVA and had a particle size of 47.9-161.1 microm. Chi-OVA dissolved readily in JP 14 first fluid, but not in JP 14 second fluid. The release of OVA from Chi-OVA was suppressed extensively in JP 14 second fluid. ER-Chi-OVA did not dissolve in JP 14 first fluid, and the release of OVA was suppressed greatly in JP 14 first and second fluids. OVA solution, Chi-OVA and ER-Chi-OVA (200 and 800 microg OVA/mouse) were administered to Balb/C mice twice at a 1-week interval. At 7 d after the second administration, plasma OVA-specific IgG and fecal OVA-specific IgA levels were measured. OVA-specific IgG tended to be enhanced in Chi-OVA and ER-Chi-OVA, but was the highest in OVA solution. OVA-specific IgA was induced significantly more efficiently by ER-Chi-OVA than the others. These suggested that ER-Chi-OVA should be possibly useful to induce an intestinal mucosal immune response. PMID- 15885939 TI - Impurity formation studies with peptide-loaded polymeric microspheres Part II. In vitro evaluation. AB - Since acylated peptide impurities were isolated from octreotide microspheres following incubation in an in vivo environment, the present investigation was undertaken to determine the dosage form dynamics responsible for facilitating acylation. In particular, microsphere batches made with poly(L-lactide) (PLA) and poly(lactide-co-glycolide) (PLGA) 85:15 were studied for in vitro drug release, mass balance relationships, mass loss behavior, hydration uptake, and solid-state stability. Furthermore, native octreotide was incubated in a varying pH stability model (heat treated lactic acid solutions 42.5%, w/w) to determine the effects of acidity on impurity formation. From a review of the experimental results, the appearance of octreotide impurities or related substances occurred with the onset of polymeric mass loss. In fact, the significant formation of acylated peptide did not appear until >90% mass loss, which was observed at 14 days. It was surmised that because of water uptake, the hydrolytic cleavage of the polymeric backbone created an acidic microenvironment to facilitate the covalent coupling of peptide with polymer. The lactic acid solution stability model corroborated with greater evidence of acylation at pH 2.25 where the presence lactoyl (+72 m/z) derivatives of octreotide were confirmed by MALDI-TOF mass spectrometry. PMID- 15885940 TI - Hypoglycemic effect of Egyptian Morus alba root bark extract: effect on diabetes and lipid peroxidation of streptozotocin-induced diabetic rats. AB - The hypoglycemic activity of the flavonoids rich fraction of 70% alcohol extract of the Egyptian Morus alba root bark (MRBF-3) was evaluated after its oral administration to streptozotocin-induced diabetic rats. Diabetes was induced by injection of 60 mg kg(-1) i.p. The administration of MRBF-3 to streptozotocin (STZ)-diabetic rats for 10 days in a dose of 200 and 400 mg kg(-1)day(-1) was not significant. However, administration of MRBF-3 for 10 days (600 mg kg(-1)day(-1)) significantly reduced the amount of the glucose from control level (379+/-9 mg/dl) to a lower level (155+/-8 mg/dl) and significantly increased the insulin level from control (10.8+/-0.3 microU/ml) to a high level (15.6+/-0.3 microU/ml). The measurement of produced lipid peroxides (expressed as the amount of thiobarbituric acid (TBA) reactive substance, nmol TBARS/ml serum) indicated antiperoxidative activity of MRBF-3. The oral administration of MRBF-3 to STZ diabetic rats significantly decreased the lipid peroxides from 6.3+/-0.8 to 5.1+/ 0.7 nmol TBARS/ml serum. The phytochemical investigation of MRBF-3 resulted in the isolation of four hydrophobic flavonoids with one or two isoprenoid groups (log P=5-9): morusin, cyclomorusin, neocyclomorusin, and kuwanon E, a 2 arylbenzofuran, moracin M, and two triterpenes, betulinic acid and methyl ursolate. The data obtained from this study revealed that MRBF-3 may protect pancreatic beta cells from degeneration and diminish lipid peroxidation. However, this is the first biological screening of the Egyptian Morus alba root bark; further future merit studies including clinical study will be necessary in order to confirm the results obtained from this study. PMID- 15885941 TI - Comparison of the antinociceptive action of crude Fuzei, the root of Aconitum, and its processed products. AB - The antinociceptive effects of crude Fuzei, the root of Aconitum carmichaeli and of Fuzei processed by three different methods were determined in mice and rats using the light tail-flick assay. A dose-dependent and significant increase in pain threshold was found at 60 min post treatment, with doses of 20-60 mg/kg crude Fuzei. The analgesic effects of processed Fuzei (20-60 mg/kg) exhibited a dose-dependent inhibition of tail-flick, but the effects were lower than those produced by crude Fuzei in the same tests. The analgesic effect of Yan-Fuzei, the salt baking product, was the most potent of the processed products and was nearly that provided by crude Fuzei. Although the concentrations of aconitine were significantly lower in the processed Fuzei than in the crude Fuzei, a higher oral LD50 was found for all of the processed Fuzei formulations. Moreover, antinociception of crude Fuzei and its processed products was attenuated but not totally blocked by naloxone at doses sufficient to block opioid mu-receptors. Furthermore, the analgesic effect of crude Fuzei and its processed products was decreased in opioid mu-receptor knockout mice, but the effect remained unaltered in mice with opioid mu-receptors, indicating that the analgesic effect of Fuzei is centrally mediated. These results demonstrate that Fuzei processed by salt baking possesses analgesic effects within a large therapeutic range, probably via a mechanism involving central opioid receptors that mediate the antinociception. PMID- 15885942 TI - Biological activity of Anthraquinones and Triterpenoids from Prismatomeris fragrans. AB - A new 1,3-dihydroxy-2-methyl-5,6-dimethoxyanthraquinone (1); six known anthraquinones, nordamnacanthal (2), damnacanthal (3), rubiadin (4), rubiadin-1 methyl ether (5), lucidin-omega-methyl ether (6), and 1-hydroxy-2-hydroxymethyl-3 methoxyanthraquinone (7); a beta-sitosterol (8); together with two known triterpenoids, beta-acetylolean-12-en-28-olic acid (9), and 3beta-O-acetyl 11alpha,12alpha-epoxyolean-28,13-olide (10) were isolated from the roots and stems of Prismatomeris fragrans. Their structures were established on the basis of spectral data. This is the first isolation of compounds 2, 6, 7, 9 and 10 from Prismatomeris genus. The isolated compounds were evaluated in antiplasmodial, antituberculosis, antifungal and anticancer cell lines tests. The bioactivity assays showed that only 9 exhibited moderate antimalarial activity, 2 and 3 exhibited antifungal activity while 2, 3, 4, 7 and 9 showed antituberculosis activity. In addition, compounds 2, 3 and 7 exhibited cytotoxicity to BC cell line while 1, 1a (the methyl ether derivative of 1), 2, 3, 4, 5, and 9 exhibited cytotoxicity to NCI-H187 cell line. PMID- 15885943 TI - Studies on the in vivo hypoglycemic activities of two medicinal plants used in the treatment of diabetes in Jordanian traditional medicine following intranasal administration. AB - The claimed hypoglycemic activities of Paronychia argentea Lam. (Caryophyllaceae) and Teucrium polium L. (Labiatae), two traditionally widely used medicinal plants in Jordan were evaluated using normoglycemic and alloxan induced hyperglycemic rabbits by intranasal administration of the plant crude extracts (10%) in a vehicle containing 5% (w/w) Pluronic F127. No significant difference was observed between the extract treated and non-treated control animals receiving only water. PMID- 15885944 TI - Antioxidant activity in Australian native sarsaparilla (Smilax glyciphylla). AB - A hot water extract of the Australian native sarsaparilla Smilax glyciphylla Sm. (Smilaceae) inhibited peroxidation of phosphatidylcholine liposomes initiated by Fe(2+)/ascorbate (IC50, 10 microg/mL) and AAPH (IC50, 33 microg/mL) in vitro. It also inhibited deoxyribose degradation and quenched chemically generated superoxide anion (IC50, 50 microg/mL). Reactivity towards ABTS (2,2'-azinobis(3 ethylbenzothiazoline 6-sulphonate) radical cation was equivalent to 48.4 mM TROLOX, the water soluble alpha-tocopherol analogue. Smilax glyciphylla is a rich source of the dihydrochalcone glycyphyllin. Given the reported level of activity it is unlikely that glycyphyllin would provide direct antioxidant protection in tissues affected by oxidative stress. However, consuming Smilax glyciphylla as a tea may be sufficient to reduce oxidative damage in the gastrointestinal tract. It is also possible that glycyphyllin is metabolised and adsorbed as phloretin, a compound with known anticancer properties. These findings indicate that further studies of the chemopreventative properties of Smilax glyciphylla is warranted. PMID- 15885945 TI - Metabolites of ephedrines in human urine after administration of a single therapeutic dose. AB - Ephedrine (EPH), pseudoephedrine (PEPH), phenylpropanolamine (PPA), methylephedrine (MEPH) and cathine are sympathomimetic amines. These drugs are commonly found in over-the-counter (OTC) cold medicines and some dietary supplements. In Taiwan, the misuse of these drugs has resulted in an increase in athletic violations. Excretion studies of the ephedrine-related drugs have been performed to better understand the metabolic yields of ephedrines in urine. After consuming a single clinical dose of each of these drugs, urine samples from volunteers (n=3 for each drug) were subjected to tert-butyl-methyl-ether (TBME) extraction and trifluoroacetic acid (TFAA) derivatization before gas chromatography-mass spectrometry (GC-MS) analysis. Most ephedrines were excreted unchanged in urine, including EPH (40.9%), PEPH (72.2%), and PPA (59.3%). However, only a relatively small amount of MEPH (15.5%) was excreted unchanged in urine. In addition, a trace amount of PPA (1.6%) and cathine (0.7%) was found to be the metabolites of EPH and PEPH, respectively. Urinary EPH, PEPH, and PPA reached peaks at 2-6h and disappeared in urine at approximately 24-48 h post administration. For MEPH, the peaks of excretion extended from 4 to 12h post administration and were undetectable at approximately 48 h. A single clinical dose of EPH (25 mg) may exceed threshold level (10 microg/mL) in sport drug testing if the urine samples are tested within approximately 8h post administration. However, a single dose of MEPH (20 mg) never reached the threshold value (10 microg/mL). PMID- 15885946 TI - Population data on 11 Y-chromosome STRs from Guine-Bissau. AB - The forensic value of Y-STR markers in Guine-Bissau was accessed by typing of 215 males. Allele and haplotype frequencies, determined for loci DYS19, DYS389-I, DYS389-II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439 and the duplicated locus DYS385, are within the limits of variation found in other populations south of the Sahara. The level of discrimination achieved is Guineans is higher than for European or other African populations with comparable data. The haplotype diversity of 0.9995 is reduced to 0.9981 when the minimal haplotype is considered thus revealing the importance of increasing the number of typed loci. PMID- 15885947 TI - Identification of inorganic anions by gas chromatography/mass spectrometry. AB - Inorganic anions were identified by using gas chromatography/mass spectrometry (GC/MS). Derivatization of the anions was achieved with pentafluorobenzyl p toluenesulphonate (PFB-Tos) as the reaction reagent and a crown ether as a phase transfer catalyst. When PFB-Br was used as the reaction reagent, the retention time of it was close to those of the derivatized inorganic anions and interfered with the analysis. In contrast, the retention time of PFB-Tos differed greatly from the PFB derivatives of the inorganic anions and the compounds of interest could be detected without interference. Although the PFB derivatives of SO4, S2O3, CO3, ClO4, and ClO3 could not be detected, the derivatives of F, Cl, Br, I, CN, OCN, SCN, N3, NO3, and NO2 were detected using PFB-Tos as the derivatizing reagent. The inorganic anions were detectable within 30 ng approximately, which is of sufficient sensitivity for use in forensic chemistry. Accurate mass number was measured for each PFB derivative by high-resolution mass spectrometry (HRMS) within a measurement error of 2 millimass units (mmu), which allowed determination of the compositional formula from the mass number. In addition, actual analysis was performed successively by our method using trial samples of matrix. PMID- 15885948 TI - Shaken baby syndrome: a biomechanics analysis of injury mechanisms. AB - Traumatic infant shaking has been associated with the shaken baby syndrome (SBS) diagnosis without verification of the operative mechanisms of injury. Intensities for SBS have been expressed only in qualitative, unsubstantiated terms usually referring to acceleration/deceleration rotational injury and relating to falls from great heights onto hard surfaces or from severe motor vehicle crashes. We conducted an injury biomechanics analysis of the reported SBS levels of rotational velocity and acceleration of the head for their injury effects on the infant head-neck. Resulting forces were compared with experimental data on the structural failure limits of the cervical spine in several animal models as well as human neonate cadaver models. We have determined that an infant head subjected to the levels of rotational velocity and acceleration called for in the SBS literature, would experience forces on the infant neck far exceeding the limits for structural failure of the cervical spine. Furthermore, shaking cervical spine injury can occur at much lower levels of head velocity and acceleration than those reported for the SBS. These findings are consistent with the physical laws of injury biomechanics as well as our collective understanding of the fragile infant cervical spine from (1) clinical obstetric experience, (2) automotive medicine and crash safety experience, and (3) common parental experience. The findings are not, however, consistent with the current clinical SBS experience and are in stark contradiction with the reported rarity of cervical spine injury in children diagnosed with SBS. In light of the implications of these findings on child protection and their social and medico-legal significance, a re-evaluation of the current diagnostic criteria for the SBS and its application is suggested. PMID- 15885949 TI - Collet-Sicard syndrome: a rare presentation of metastatic prostate adenocarcinoma. AB - Metastatic spread of prostate adenocarcinoma to the temporal bone is rare. Involvement the jugular foramen may result in Collet-Sicard syndrome. This extremely uncommon clinical entity is characterized by paralysis of the lower four cranial nerves. A case of metastatic prostate adenocarcinoma involving the temporal bone causing a Collet-Sicard syndrome associated with an ipsilateral lower motor facial palsy and a mixed sensorineural and conductive hearing loss is presented. This case highlights the potential of prostate adenocarcinoma to cause symptoms referable to the temporal bone region and histopathologic analysis of biopsy material should include immunohistochemical staining for prostate specific antigen. PMID- 15885950 TI - The relationship between esophagoscopic findings and total acid reflux time below pH 4 and pH 5 in the upper esophagus in patients with laryngopharyngeal reflux disease (LPRD). AB - OBJECTIVE: : To investigate the relationship between esophagoscopic findings and total acid reflux time in the upper esophagus in the patients of laryngopharyngeal reflux disease (LPRD), and to evaluate whether pH<5 holding time is appropriate for diagnosing LPRD. METHOD: : Esophagoscopy and 24-h pH monitoring in the upper esophagus were performed in 29 patients suspected of LPRD and 19 healthy volunteers participated as controls. The total acid reflux times below pH 4 and 5 in the upper esophagus were compared between the LPRD group and the controls. The LPRD group was divided into LPRD with reflux esophagitis (RE) and LPRD without RE, and the total acid reflux times below pH 4 and 5 of these two groups were compared. RESULTS: : Eight of the 29 (27.6%) LPRD patients had reflux esophagitis. The LPRD group showed a significantly longer acid reflux time than the control group at pH below 5 (P=0.009) but not at pH below 4 (P=0.48). The LPRD with RE had significantly longer acid reflux times below both pH 4 and 5 in the upper esophagus compared to those of the LPRD without RE. The P values using a cut-off value of pH 5 were smaller than that of pH 4 in all statistical results. CONCLUSION: : The LPRD patients showed significantly longer acid reflux time in the upper esophagus than the control group. In the LPRD group, the LPRD with RE patients had more frequent acid exposure in the upper esophagus than the LPRD without RE. A pH level of 5 could be appropriate as a cutoff level for the diagnosis of LPRD in upper esophagus pH monitoring. PMID- 15885951 TI - Plantago lanceolata (English plantain) pollinosis in Japan. AB - OBJECTIVE: The objective was to ascertain the prevalence of Plantago lanceolata (English plantain) pollinosis in Japan. METHODS: A total of 160 patients with allergic rhinitis were examined at the Hokkaido University Hospital Otolaryngology Clinic between January 2002 and December 2003. We investigated the frequency of P. lanceolata antigen-specific immunoglobulin E (IgE) antibody positive serum using the radioallergosorbent test (RAST), the P. lanceolata pollen counts in the Sapporo area and the clinical symptoms of patients with allergic rhinitis caused by this pollen. RESULTS: P. lanceolata pollen was dispersed throughout the Sapporo area from mid-May to early September, peaking in the latter half of May. The airborne pollen count showed little variation between years. The P. lanceolata RAST-positive rate was 12.8%, which was close to those for ragweed and sagebrush. All subjects showed overlapping antigen sensitization, that is, none displayed sensitization (by RAST) exclusively to P. lanceolata. The onset of subjective symptoms peaked in June and the symptoms worsened from April to October. CONCLUSION: P. lanceolata was dispersed over a long period, from mid May to early September. The P. lanceolata RAST-positive rate (12.8%) was similar to those for sagebrush and ragweed, which are dispersed during the fall. These results suggest that P. lanceolata is an important causative agent of pollinosis in Japan. PMID- 15885952 TI - Voice restoration by primary insertion of indwelling voice prosthesis following circumferential pharyngolaryngectomy with free jejunal graft. AB - BACKGROUND: Voice restoration after circumferential pharyngolaryngectomy (CPL) with free jejunal graft remains a difficult problem to solve. Few reports have analyzed the success rate and complications following primary insertion of indwelling voice prostheses during CPL with free jejunal graft. PATIENTS AND METHODS: Eight patients who underwent CPL with free jejunal graft had a Groningen voice prosthesis inserted as a tracheoesophageal (TE) shunt at the time of oncological surgery. A 10-point scale was used to assess each patient's speech intelligibility. Complications following the voice prosthesis insertion were also analyzed. RESULTS: Six of the eight patients (75%) achieved excellent speech intelligibility and the remaining two patients (25%) were judged as moderate. Six of the eight patients (75%) used the TE shunt as their major means of daily communication. Leakage through or around the prosthesis, which occurred in six (75%) patients, was the most frequent prosthesis-related complication. CONCLUSIONS: This safe and reliable technique can be effective in improving the quality of life in selected patients undergoing CPL. PMID- 15885953 TI - Expression of full-length Cochlin p63s is inner ear specific. AB - OBJECTIVE: The COCH gene mutated in DFNA9, murine an autosomal dominant hereditary hearing impairment, encodes Cochlin. Cochlin is also suggested to be the self-antigen of autoimmune sensorineural hearing loss. We previously reported that Cochlin constitutes 70% of the inner ear proteins and is classified into three types of isoform, p63s, p44s, and p40s. To study the specificity of expression of Cochlin isoforms in various organs, here we have investigated expression of the COCH gene at both the transcriptional and translational level. METHODS: COCH gene expression was studied by RT-PCR and Southern blot analysis. Cochlin isoforms were studied by Western blot analysis using an isoform specific antibody. RESULTS: At the transcriptional level, COCH mRNA was detected only in the inner ear by RT-PCR. Southern blot analysis of RT-PCR products detected a high level of COCH mRNA in the inner ear, lower level in spleen, and very low levels in the cerebrum, cerebellum/brain stem, eye, liver and kidney. At the translational level, Western blot analysis showed that a set of isoform, p63s, p44s, and p40s was detected at high levels only in the inner ear. In contrast, multiple proteins were detected at much lower levels in other organs tested. Notably, full-length Cochlin p63s was detected only in the inner ear. CONCLUSION: Our findings demonstrate that the COCH gene is expressed preferentially in the inner ear and that expression of full-length Cochlin p63s is specific to the inner ear. These results will be central to understanding the function of Cochlin and its role in the pathophysiology of DFNA9. PMID- 15885954 TI - [Anal condyloma prevalence among HIV infected patients]. AB - BACKGROUND: In the general HIV-infected population, there are few data on the prevalence and risk factors for anal condyloma, precursor lesions for anal cancer. METHODS: Screening for perianal and endoanal condyloma with an anoscopy was systematically proposed to 516 consecutive outpatients, followed in a university hospital in Paris. For each point, HIV characteristics and sexual behaviors assessed through semi-directive questionnaire were collected. FINDINGS: The 473 (92%) examined patients, consisted of 200 homosexual men, 123 heterosexual men, 150 women; 76% were receiving HAART, HIV-RNA was<50 copies/ml in 60%, mean (+/-SD) CD4 cell count were 484 (+/-274)/mm(3). Overall, 108 (23%) pts had histologically-confirmed anal condyloma (36, 15 and 11% of the respective populations), including 51 (47%) pts with only endoanal localisation. Intraepithelial neoplasia of grade I was noted in 59 patients, of grade II in 10 and of grade III in 2 and an invasive endoanal cancer in 1. In multivariate regression analysis, condyloma independent risk factor were history of gonococcia or syphilis (OR=0.54 (0.29-0.99)), and history of previous anal condyloma (OR=2.05 (1.07-3.92) in homosexual men, history of previous penis condyloma (OR=26.8 (2.3-309.6), and unprotected sexual intercourse (OR=7.5 (2.1-26.3)) in heterosexual men and CD4 cell count below 200/mm(3), (OR=8.9 (1.5-51.6)), receptive anal intercourse (OR=6.7 (1.7-25.8)) and history of previous anal condyloma (OR=25.4 (3.4-188.2)) in women. INTERPRETATION: In the HAART era, systematic screening revealed a high rate of anal condyloma in all HIV positive pts (not only in homosexual men). Anal examination should be proposed systematically to all HIV-infected patients. PMID- 15885955 TI - [Survey of foot and mouth disease in France]. AB - The Foot and Mouth disease is considered a relatively mild disease in susceptible animals but it has a considerable economical impact in France and worldwide due to the huge economical losses that it generates. A system for the prevention of the disease was developed thirty years ago and has since been continuously improved. The current system for the prevention of FMD is based on training and information of all those involved in the surveillance of susceptible animals, over all the national territory, and on the control of imported animals and animal products. This system, developed in close collaboration with the European Commission and the other EU member states, allows control measures to be implemented almost instantaneously, if a case is suspected to avoid spreading of the disease. PMID- 15885956 TI - [European surveillance of tuberculosis: description of the network and recent results]. AB - A network for the surveillance of tuberculosis covering the WHO European Region was set up in 1996. It aims to improve the contribution of surveillance to TB control, promoting standardised methods to compare countries. Standardized individual or aggregate data on notified TB cases is provided annually from national surveillance institutions. In the enlarged European Union (EU), overall TB notification rates decreased in recent years (14/100 000 in 2002), but leveled off in some countries including France and the United Kingdom, partly due to increasing numbers of patients from high TB incidence areas (30% in 2002). Multidrug resistance (MDR) was much more frequent in the Baltic States (21% in 2002) than in other countries (1.7%; range: 0-3.7%). Treatment of new pulmonary TB cases notified in 2001 was completed within 12 months in 74% of cases, ongoing in 3%, interrupted due to death in 7%, and interrupted for other reasons or unknown in 15%. In the Balkans and Turkey, notification rates ranged between 20 and 62 per 100 000 in 2002, and decreasing or stable except for Romania (153 in 2002). In the East (former Soviet Union), TB surveillance data, although incomplete, shows an increasing incidence (97/100 000 in 2002), high prevalence of primary MDR (14% in Kazakhstan), and frequent treatment failures (10% in new cases). At the time of EU enlargement, European TB surveillance covering the whole WHO European Region should continue, with support of TB control in the East as a regional priority. PMID- 15885957 TI - Estradiol and soy extract increase the production of new cells in the dentate gyrus of old rats. AB - In young rodents, estradiol increases cell proliferation in the dentate gyrus of the hippocampus. However, it is unknown if the old brain retains this response to estradiol. Here we assessed the generation of new cells in the dentate gyrus of old rats after administration of estradiol or a soy extract, since soy is used as an alternative to hormonal replacement therapy in postmenopausal women. In a first experiment, 12-month-old animals were ovariectomized and studied at 14, 18 or 22 months of age. The production of new cells, assessed by the incorporation of bromodeoxyuridine (BrdU), was similar in 14- and 18-month-old rats. However, there was a significant reduction in the number of BrdU-immunoreactive cells at 22 months of age. In a second experiment, 22-month-old ovariectomized animals were treated for 10 weeks with a weekly s.c. injection of 150 microg estradiol valerianate or with 60 mg/kg per day soy extract added to the drinking water. Both treatments increased significantly the production of new cells in the dentate gyrus. These findings indicate that the brains of old rats retain the ability to increase the production of new cells in response to estradiol and soy extracts. PMID- 15885958 TI - Percutaneous management of parapneumonic effusions. AB - Parapneumonic effusions continue to be a significant source of morbidity and mortality. Treatment at earlier stages before fibrous peel and loculations occur has a much better prognosis. Using image guidance, addition of intracavitary fibrinolytic instillation, close follow-up with drainage of residual or new collections are some of the other factors that improve prognosis. In this article, we discussed treatment strategies, percutaneous management of parapneumonic effusions, its alternatives and results. PMID- 15885959 TI - Radiologic imaging and percutaneous treatment of pelvic lymphocele. AB - Pelvic lymphocele, also known as lymphocyst, is a cystic structure caused by lymphatic injury usually secondary to pelvic lymphadenectomy and renal transplantation. Lymphoceles can cause morbidity and rarely mortality by compression of adjacent structures and infectious complications. This review discusses etiology and treatment options for pelvic lymphoceles including surgical and percutaneous methods with emphasis on percutaneous techniques particularly in conjunction with sclerotherapy. Percutaneous catheter drainage with sclerotherapy procedure with various sclerosing agents is described in detail. Ethanol, povidone-iodine, tetracycline, doxycycline, bleomycin, talc and fibrin glue can be used as sclerosing agents. Combination of sclerosing agents to percutaneous catheter drainage significantly improves success rate in the treatment of pelvic lymphoceles. Infected lymphoceles are usually treated solely with percutaneous catheter drainage. Percutaneous treatment can be tailored according to volume of lymphoceles. We generally prefer single session sclerotherapy and 1 day catheter drainage in lymphoceles less than 150 mL, and larger ones are treated by multi-session sclerotherapy until daily drainage decreases below 10 mL. Percutaneous treatment preferably with sclerotherapy should be considered as the first-line treatment modality for pelvic lymphoceles due to its effectiveness, widespread applicability on an outpatient basis, ease of procedure and low complication rate. PMID- 15885960 TI - Molecular imaging of apoptosis in cancer. AB - Apoptosis plays an important role in cancer. Mechanisms hindering its action are implicated in a number of malignancies. Also, the induction of apoptosis plays a pivotal role in non-surgical cancer treatment regimes such as irradiation, chemotherapy, or hormones. Recent advanced in imaging science have made it now possible for us to detect and visualize previously inaccessible and even unrecognized biological phenomena in cells and tissue undergoing apoptosis in vivo. Not only are these imaging techniques painting an intriguing picture of the spatiotemporal characteristics and metabolic and biophysical of apoptosis in situ, but they are expected to have an ever increasing impact in preclinical testing and design of new anticancer agents as well. Rapid and accurate visualization of apoptotic response in the clinical settings can also be of significant diagnostic and prognostic worth. With the advent of molecular medicine and patient-tailored treatment options and therapeutic agents, such monitoring techniques are becoming paramount. PMID- 15885961 TI - Feeding-unrelated factors influencing the plasma leptin level in ruminants. AB - The triglyceride content of lipid depots associated with the current feeding level is the primary determinant of leptin gene expression and the circulating leptin level. In laboratory rodents and primates the plasma leptin is influenced also by the age, gender and physiological status (puberty, pregnancy, lactation, postpartum period), and by the health condition such as sepsis due to Gram negative (GN) bacteria. Some pathologic conditions with intensive cytokine release evoke an increase in plasma leptin, which is thought to depress the subsequent feed intake. However, the effect of these secondary factors may be species-dependent, with still unknown clinical relevance in ruminants. In our ovine and bovine models plasma leptin increased after castration and dexamethasone treatment, decreased after experimental administration of synthetic androgens in castrated rams, but remained unchanged throughout the ovarian cycle and after ovariectomy. The circulating leptin level increased temporarily during synthetic progestin (fluorogestone) treatment in ewes, but similar changes were not seen in progesterone-supplemented ewes and norgestomet-treated cows. In a second trial on dairy cows we wanted to study whether elevated plasma leptin levels are induced by experimental endotoxin mastitis, or by natural outbreak of GN mastitis and puerperal metritis. Experimental endotoxin mastitis resulted in some-hour elevation in cortisol and insulin, with a simultaneous decrease in IGF I and thyroid hormones. In the first 14 days of lactation GN mastitis induced the same endocrine alterations as the experimental endotoxin challenge, but in natural cases these changes varied within a wider range, and were more protracted and robust. Cows with puerperal metritis had more obvious catabolic changes in the early weeks of lactation, than their healthy counterparts. However, both mastitis and puerperal metritis failed to increase the circulating leptin level, showing that in cows the plasma leptin is not responsible for the anorexia associated with these inflammatory diseases. PMID- 15885962 TI - Regulation of pituitary somatotroph differentiation by hormones of peripheral endocrine glands. AB - Anterior pituitary somatotroph differentiation occurs during chick embryonic and rat fetal development. A number of findings support the hypothesis that differentiation of these growth hormone (GH) producing cells in the chick and the rat is regulated by adrenal glucocorticoids and thyroid hormones. Somatotroph differentiation can be induced in cultures of chick embryonic and rat fetal pituitary cells with adrenal glucocorticoids and this effect can be modulated by concomitant treatment with thyroid hormones. Plasma levels of thyroid hormones, corticosterone and adrenocorticotropic hormone increase during development, consistent with the ontogeny of somatotrophs. Treatment of chick embryos or rat fetuses with glucocorticoids in vivo induces premature somatotroph differentiation, indicating that the adrenal gland, and ultimately anterior pituitary corticotrophs, may function to regulate pituitary GH cell differentiation during development. Administration of thyroid hormones in vivo also increases somatotrophs prematurely, and administration of the thyroid hormone synthesis inhibitor methimazole inhibits somatotroph differentiation in vivo, suggesting that endogenous thyroid hormone synthesis contributes to normal somatotroph differentiation. Our working model for the regulation of somatotroph differentiation during normal development includes modulation by elements of the hypothalamo-pituitary-adrenal and hypothalamo-pituitary-thyroid axes. Additional research is reviewed defining the mechanism of action for these peripheral hormones in induction of pituitary GH gene expression during development. PMID- 15885963 TI - Can farm animals help to study endocrine disruption? AB - The phenomenon of endocrine disruption can be regarded as part of the disciplines of toxicology and environmental toxicology. These two disciplines have generated guideline protocols on how various effects of chemicals should be tested as a basis for regulatory decisions. These protocols almost exclusively involve laboratory rodents and the data obtained are then used for human risk assessment. Would it be justifiable, then, to introduce or promote the use of other species in these test protocols? There are, at any rate rationales for studying effects in species other than laboratory rodents: (1) other species may better mimic the human system; (2) they may in some cases be more useful for studying a certain mechanism or phenomenon; (3) they may highlight the diversity of effects or sensitivity between species. However, there are at least two basic criteria that must be met for a species before it can be introduced in this context: (a) we must have a good understanding of the physiological system to be studied; and (b) we must have a number of tools to study effects on this system. When it comes to the reproductive system--regarding which most endocrine disruption has been reported--farm animals are second only, or in some respects superior, to laboratory rodents with respect to these criteria. This review gives examples of how farm animals can be of use in the study of endocrine disruption with a focus on the author's own data from studies in the pig. PMID- 15885964 TI - [Involvement of anaesthesiologists in surgery offices: toward a change of attitude?]. PMID- 15885965 TI - [Epilepsy in patients suffering from severe subarachnoid haemorrhage]. PMID- 15885966 TI - [Cerebral oxidative stress: are astrocytes vulnerable to low intracellular glutamate concentrations? Consequences for neuronal viability]. AB - This review describes reactive oxygen species (ROS), their production and effects on crucial biological molecules, the different lines of defense against oxidative stress, with particular attention to glutathione, the main antioxidant in the brain, which neuronal synthesis seems to be dependent on astrocytic precursors. It also focuses on the different ways by which glutamate may induce oxidative stress in the brain. The different mechanisms leading to ROS production, activated during the excitotoxic cascade, are described. Oxidative glutamate toxicity is also briefly described. A novel form of oxidative glutamate toxicity by depletion of transported glutamate that we recently evidenced is detailed. This toxicity induced by pharmacological reversal of glutamate transport, which mimics glutamate transport reversal occurring in ischemia, involves glutathione depletion and oxidative stress, leading to delayed death of cultured striatal astrocytes differentiated by dibutyryl-cAMP, probably through apoptotic processes. Evidence suggesting that this oxidative glutamate toxicity by depletion of transported glutamate is very likely occurring in vivo and its consequences on neuronal survival are discussed. PMID- 15885967 TI - [SRLF experts recommendations: Indicators of volume resuscitation during circulatory failure]. PMID- 15885968 TI - [Vasospasm after severe subarachnoid haemorrhage: diagnosis and treatment]. PMID- 15885969 TI - [Severe subarachnoid haemorrhage: treatment of rebleeding and of an intracerebral haematoma]. PMID- 15885970 TI - Crack cocaine-induced long QT interval syndrome in a parturient with recreational cocaine use. PMID- 15885971 TI - [Anaesthetic management of the patient with acute intracranial hypertension]. AB - Transcranial Doppler and, if possible, measurement of intracranial pressure (ICP) allow preoperative diagnosis of acute intracranial hypertension (ICH) after brain trauma. The main goal of the anaesthesiologist is to prevent the occurrence of secondary brain injuries and to avoid cerebral ischaemia. Treatment of high ICP is mainly achieved with osmotherapy. High-dose mannitol administration (1.4 to 2 g/kg given in bolus doses) may be considered a better option than conventional doses, especially before emergency evacuation of a cerebral mass lesion. Hypertonic saline seems as effective as mannitol without rebound effect and without diuresis increase. Haemostasis should be normalized before neurosurgery and invasive blood pressure monitoring is mandatory. For anaesthesia induction, thiopental or etomidate may be used. In case of ICH, halogenated and nitrous oxide should be avoided. Until the dura is open, mean arterial pressure should be maintained around 90 mmHg (or cerebral perfusion pressure around 70 mmHg). If a long-lasting (several hours) extracranial surgery is necessary, ICP should be monitored and treatment of ICH should have been instituted before. PMID- 15885972 TI - [Conventional and diffusion magnetic resonance imaging in the acute phase of severe traumatic brain injury]. AB - Neuro-imaging is essential for the initial evaluation and subsequent control in the acute stage of severe head injury. In these indications tomodensitometry (TDM) has a pivotal role. Despite the well recognized contribution of magnetic resonance imaging (MRI) to the investigation of most of acute neurological pathologies, MRI is not still a routine procedure for the initial investigation of patients with acute head injury. The superiority of morphological and functional MRI on TDM in this indication is discussed. PMID- 15885973 TI - [Cardiovascular and pulmonary consequences of severe subarachnoid haemorrhage]. PMID- 15885974 TI - [Medical treatment of an important trachea injury after tracheal intubation]. AB - Tracheal injuries are rare but life threatening complications of anaesthesia. Two therapeutic strategies are presently proposed: a non-surgical strategy for small injuries and a surgical strategy for larger ones. This case report presented the non-surgical therapeutic strategy of a large tracheal injury. This strategy was discussed according to other published reports. PMID- 15885975 TI - [Severe subarachnoid haemorrhage: natremia and renal function]. PMID- 15885976 TI - [Fascia iliaca block for femoral bone fractures in prehospital medicine]. AB - OBJECTIVE: The aim of this study was to assess the feasibility and efficacy of fascia iliaca compartment bloc (FIB) in prehospital care performed by emergency physicians. STUDY DESIGN: Prospective observational study. PATIENTS AND METHODS: Fifty-two patients victim of a femoral bone fracture were included consecutively. All FIB had been performed by emergency physicians trained to the technique. Lidocaine 1.5% with epinephrine has been used. Block efficacy was assessed by testing sensitive block in the anterior, lateral and medial part of the thigh. Pain levels were noted using a simplified numeric scale (SNS). RESULTS: No complete was reported. Ninety-four percent of blocks were successful. SNS values significantly decrease 10 minutes after block performance. CONCLUSION: Emergency medicine physicians trained to the technique can perform FIB with a high success rate. PMID- 15885977 TI - A C1q-binding 40 kDa porin from Aeromonas salmonicida: cloning, sequencing, role in serum susceptibility and fish immunoprotection. AB - A 40 kDa C1q-binding outer membrane protein from Aeromonas salmonicida was identified by direct-binding assay with biotinylated C1q, a subcomponent of the complement classical pathway component C1. The 40 kDa porin structural gene from the A450 A. salmonicida typical strain (A+:O+) was cloned in Escherichia coli and sequenced. The amino acid sequence of the 40 kDa A. salmonicida porin, its ability to bind C1q in an antibody independent process, and its immunological cross-reaction with the A. hydrophila AH-3 porin II, allow us to determine the role of this protein in serum susceptibility. Furthermore, we obtained defined A. salmonicida 40 kDa porin insertion mutants in serum sensitive or resistant strains, and we complemented these mutants with a plasmid harboring only the 40 kDa porin gene from A. salmonicida A450 in order to define its role as an important surface molecule involved in serum susceptibility and C1q binding. Similar complementation was obtained using the A. hydrophila AH-3 porin II gene. The 40 kDa porin gene and/or protein was present in all the A. salmonicida typical or atypical strains tested. Furthermore, the A. hydrophila AH-3 porin II seems to be an important molecule for fish immunoprotection against either A. salmonicida or A. hydrophila strains. PMID- 15885978 TI - An experimental model for the determination of immunomodulating effects by volatile compounds. AB - An in vitro exposure system was developed to enable simultaneous exposure of primary cells or cell lines to defined concentrations of volatile organic compounds (VOC) without the necessity of a constant-flow exposure system. Toluene was used as model VOC and administered via the gas phase. CD3/CD28-stimulated human peripheral blood mononuclear cells (PBMC) were used as indicator cells. Vitality/proliferation of PBMC was tested using the MTT assay and their functional reactivity using cytokine ELISA for interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-13, and tumor-necrosis-factor-alpha (TNF-alpha). Chemical analysis using headspace gas chromatography confirmed that this new method guaranties reproducible VOC exposure (R2 = 0.995 for the correlation between external toluene concentration and toluene in the cell culture). While cytotoxic effects were not observed, dose-dependent toluene effects on functional reactivity of PBMC were found. The secretion of IFN-gamma, IL-4, and IL-13 was inhibited at concentrations of 72.5 g/m3 and above, whereas the TNF-alpha production was increased. Since the presented in vitro model ensures toluene exposure in concentrations comparable to the real situation, and allows the investigation of dose-dependent immunomodulatory toluene effects in concentrations without cytotoxicity, this method first described here is introduced as useful tool in analysis of VOC-triggered effects on immune cells. PMID- 15885979 TI - Depression, anxiety and neuropsychological test scores of candidates for coronary artery bypass graft surgery. AB - The effect of depression and anxiety upon neuropsychological test scores of candidates for coronary artery bypass graft (CABG) surgery was examined. Sixty patients were administered the Beck Depression Inventory II and the State-Trait Anxiety Inventory, along with a battery of neuropsychological tests. Regression analyses were conducted in which the neuropsychological test scores were predicted using age, education, depression, anxiety, and combined depression anxiety scores. While age and education were significant predictors of several neuropsychological test measures, no significant regression analysis results were obtained for the depression, anxiety and combined depression-anxiety scores. Similarities and discrepancies between this research and previous studies are discussed. It appears that low levels of preoperative depression and anxiety states do not affect neuropsychological functioning among CABG candidates. PMID- 15885980 TI - Localized breakdown in linkage disequilibrium does not always predict sperm crossover hot spots in the human MHC class II region. AB - To investigate the relationship between meiotic crossover hot spots and block like linkage disequilibrium (LD), we have extended our high-resolution studies of the human MHC class II region to a 90-kb segment upstream of the HLA-DOA gene. LD blocks in this region are not as well defined as in the neighboring 210-kb DNA segment but do show two regions of LD breakdown in which coalescent analysis indicates substantial historical recombination. Sperm crossover analysis of one region revealed a novel localized hot spot similar in intensity and morphology to most other MHC hot spots. Crossovers at this hot spot are not obviously affected by a large insertion/deletion polymorphism near the hot spot. The second region of LD breakdown, within the DPB1 gene, shows an extremely low level of sperm crossover activity and does not contain a sperm crossover hot spot. These results highlight the complexity of LD patterns and the importance of experimentally verifying crossover hot spots. PMID- 15885981 TI - Stereotypy I: a review of behavioral assessment and treatment. AB - In this paper, we review definitional issues related to stereotypy, behavioral interpretations of stereotypy, procedures for determining operant function(s) of stereotypy, and behavioral interventions for stereotypy. In general, a preponderance of the assessment literature suggests that most forms of stereotypy are maintained by automatic reinforcement. Review of the treatment literature suggests that antecedent (e.g., environmental enrichment) and consequent (e.g., differential reinforcement of alternative behavior) interventions produce at least short-term reductions in stereotypy. Suggestions for further assessment and treatment of stereotypy are provided. PMID- 15885982 TI - Pre- and postnatal expressions of NMDA receptors 1 and 2B subunit proteins in the normal rat cortex. AB - The NMDA receptor is one of the ionotropic glutamate receptors essential for excitatory neurotransmission and synaptic plasticity. The present study examined the ontogenies of NMDAR1 and NMDAR2B during development (embryonic days: (E) 14, 15, 16, 17 and postnatal days: (P) 0, 5, 10, 20, and 60). The immunoreactivities of both NMDAR1 and NMDAR2B were visualized and semi- quantified simultaneously using batch-matched procedures during the developmental period. In addition, the heteromeric coassembly of NMDAR1 (NR1) and NMDAR2B (NR2B) was analyzed throughout development using immunocoprecipitation blots. Differential developmental expressions were observed between the two NMDA receptor subunits. Although both NR1 and NR2B were already expressed on cortical neurons at E14, NR1 immunoreactivity was lower than NR2B but increased up to P20 then was lower at the last sampling day (P60). By comparison, the high NR2B immunoreactivity at E14 increased until P5, and until the next sample day (P60). Despite the early embryonic protein expressions in prenatal cortex both NR1 and NR2B immunocoprecipitation analyses showed that NR1-NR2B coassembly was not initially detectable until birth (P0), and increased up to P20. This suggests that the physically interactive NMDAR1-2B complex may contribute to the plasticity of early postnatal development characterized by axonal dendritic synaptogenesis. PMID- 15885983 TI - What are the effects of group cognitive behaviour therapy for voices? A randomised control trial. AB - BACKGROUND: Little evidence exists for the effects of psychological treatment on voices even though it is clear that CBT does affect delusions and symptoms overall. This study tested whether a group based on cognitive behavioural principles could produce beneficial effects on hallucinations. AIM: To test the effectiveness of group CBT on social functioning and severity of hallucinations. METHOD: Participants were included if they had a diagnosis of schizophrenia and experienced distressing auditory hallucinations (rated on the PANSS). They were randomly allocated to group CBT (N = 45) or a control group who received treatment as usual (N = 40). The two main outcomes were social functioning as measured by the Social Behaviour Schedule and the severity of hallucinations as measured by the total score on the Hallucinations Scale of PSYRATS. Assessments were carried out at baseline, 10 weeks (post therapy) and 36 weeks (six months following therapy). RESULTS: Mixed random effects models revealed significant improvement in social functioning (effect size 0.63 six months after the end of therapy). There was no general effect of group CBT on the severity of hallucinations. However, there was a large cluster effect of therapy group on the severity of hallucinations such that they were reduced in some but not all of the therapy groups. Improvement in hallucinations was associated with receiving therapy early in the trial and having very experienced therapists (extensive CBT training which included expert supervision for a series of individual cases for at least a year following initial training). CONCLUSION: Group CBT does improve social functioning but unless therapy is provided by experienced CBT therapists hallucinations are not reduced. PMID- 15885984 TI - Pathogenesis of systemic sclerosis: altered B cell function is the key linking systemic autoimmunity and tissue fibrosis. AB - Systemic sclerosis (SSc) is characterized by autoimmunity and tissue fibrosis. There is a close association between specific autoantibodies and clinical features in patients with SSc. A number of studies have demonstrated that various cytokines, such as transforming growth factor-beta, modulate the synthesis of extracellular matrix by fibroblasts. However, it is not clear as to how autoimmunity and tissue fibrosis interact with each other. Recent studies have revealed that B cells play a critical role in various systemic autoimmune disorders. CD19 is a central regulator of B cell signaling threshold, and B cells from SSc patients exhibit an increased expression of CD19 that induces SSc specific autoantibody production in transgenic mice. Furthermore, SSc patients have intrinsic B cell abnormalities characterized by decreased but activated memory B cells, which is possibly due to CD19 overexpression. Similarly, B cells from a tight-skin mouse, a model of SSc, show augmented CD19 signaling and chronic B cell activation. Remarkably, CD19 loss results in inhibition of chronic B cell hyper-reactivity and elimination of autoantibody production, which is associated with improvement in skin fibrosis and a parallel decrease in IL-6 production by B cells. Therefore, augmented cytokine production by B cells is a potential candidate for the induction of skin sclerosis. Alternatively, B cells may influence tissue fibrosis by regulating T cell activation and cytokine production through their antigen-presenting and co-stimulatory abilities. Thus, altered B cell function may result in tissue fibrosis, as well as autoimmunity, in SSc. PMID- 15885985 TI - Establishment of tyrosinase sequence database in normally pigmented Indians and Japanese for rapid determination of novel mutations. AB - BACKGROUND: Many mutations of the tyrosinase gene have been reported in oculocutaneous albinism type I (OCA1) patient. In the future, a greater number of novel mutations will be found as the search for pathological mutations in the tyrosinase genes of OCA patients from various ethnic origins. For rapid determination in future whether an observed mutation is a polymorphism or a novel pathological one, sequence databases of the gene of various ethnic people are needed. OBJECTIVE: We established a sequence database of the tyrosinase gene of Japanese as well as Indian people. METHOD: We collected DNA from 109 Japanese and 103 Indians with normal pigmentation and analyzed their tyrosinase gene using a direct sequencing method. RESULT: The database shows an apparent difference between the two ethnic groups in polymorphisms of the tyrosinase gene namely, Q402 allele, Y192 allele and IV2+24 insT were found in the Indian population, but not in the Japanese. On the other hand, some Japanese had IV2-21 insT but none of the Indians did. The database supports the notion that the tyrosinase gene evolved and extended separately in the two ethnic groups. And the developing database confirmed that the reported mutations causing Indian and Japanese OCA were not among the polymorphisms in the database, which conversely gives genetical proof of the "genuine" pathological mutations. CONCLUSION: Eventually, the sequence database we established will contribute to demonstrating novel mutations of albinism in Indians and Japanese. PMID- 15885986 TI - Piperacillin-sulbactam versus piperacillin-tazobactam: a multicentre, randomised, single-blind, controlled clinical trial. AB - The objective of this study was to compare the efficacy and safety of piperacillin-sulbactam (PIP-SBT) and piperacillin-tazobactam (PIP-TAZ) in the treatment of bacterial respiratory and urinary tract infections. A randomised, single-blind, controlled clinical trial was performed. Differences in clinical efficacy, bacteriology and safety between the two groups were subjected to statistical analysis, including intent-to-treat (ITT) analysis. A total of 215 cases were enrolled, with 203 complete cases (99 PIP-SBT, 104 PIP-TAZ). A total of 209 cases (103 PIP-SBT, 106 PIP-TAZ) were included in the ITT analysis and a total of 212 cases (104 PIP-SBT, 108 PIP-TAZ) were included in the safety analysis. Overall efficacy rates of PIP-SBT and PIP-TAZ were 93.2% and 93.4%, respectively. Overall bacterial eradication rates of the two groups were 95% and 97.59%, respectively. Among the PIP-SBT group, eight patients (7.69%) had adverse events, including four probable drug-related events. Among the PIP-TAZ group, nine patients (8.33%) had adverse events, including one definitely drug-related and four probable drug-related events. All differences between the two groups were insignificant. PIP-SBT could be a suitable replacement for PIP-TAZ in the therapy of community-acquired respiratory and urinary tract infections caused by beta-lactamase-producing bacterial isolates. PMID- 15885987 TI - Steady-state serum and intrapulmonary pharmacokinetics and pharmacodynamics of tigecycline. AB - The steady-state serum and intrapulmonary pharmacokinetic and pharmacodynamic parameters of tigecycline were determined after intravenous administration in 30 subjects. Tigecycline was administered as a 100mg loading dose followed by six 50mg doses given every 12h and was measured using HPLC/mass spectrometry. Ratios of tigecycline maximum serum concentration and area under the serum concentration time curve to 90%-minimum inhibitory concentrations (C(max)/MIC(90); AUC/MIC(90)), and percentage time above MIC(90) were calculated for common respiratory pathogens (Streptococcus pneumoniae, Chlamydia pneumoniae, Mycoplasma pneumoniae, Moraxella catarrhalis and Haemophilus influenzae). The C(max) (mean+/ S.D.), AUC and half-life for serum were 0.72+/-0.24 microg/mL, 1.73+/-0.64 microg*h/mL and 15.0+/-1.10h; for lung epithelial lining fluid (ELF) the values were 0.37 microg/mL, 2.28 microg*h/mL and 39.1h; and for alveolar cells (AC) were 15.2 microg/mL, 134 microg*h/mL and 23.7h. Tigecycline was concentrated in AC: C(max)/MIC(90) ratios ranged from 30.4 (H. influenzae) to 507 (S. pneumoniae); AUC/MIC(90) ratios ranged from 268 (H. influenzae) to 4467 (S. pneumoniae); and percentage dose interval above MIC(90) was 100% for the five respiratory pathogens. The C(max)/MIC(90), AUC/MIC(90) ratios, T>MIC(90) and extended serum and intrapulmonary half-lives following the regimen used in this study are favourable for the treatment of tigecycline-susceptible pulmonary infections. PMID- 15885988 TI - Activity of RBx 7644 and RBx 8700, new investigational oxazolidinones, against Mycobacterium tuberculosis infected murine macrophages. AB - The decision to develop a new chemical entity should not only be based on its ability to inhibit multidrug-resistant tuberculosis (MDR-TB) strains but also on its ability to enter macrophages and be active against intracellular bacteria. RBx 7644 and RBx 8700, two novel extended spectrum oxazolidinones, were investigated for their activity against sensitive and MDR isolates of Mycobacterium tuberculosis and for activity against bacteria within a macrophage cell line. RBx 8700 showed excellent in vitro activity against sensitive as well as MDR M. tuberculosis strains with MIC(50) and MIC(90) values of 0.032 and 0.25mg/L (sensitive) and 0.25 and 1.0mg/L (MDR) strains. The corresponding MIC(50) and MIC(90) values of RBx 7644, linezolid, rifampicin and isoniazid were 8 and 16; 32 and 64; 64 and 64; 64 and 64 mg/L, respectively. RBx 8700 and rifampicin were bactericidal at 0.5 and 0.25mg/L when tested intracellularly whereas linezolid reduced the count by 100-fold at a concentration of 8 mg/L. In combination studies with standard antimycobacterial drugs, RBx 8700 did not show any antagonistic effect. PMID- 15885989 TI - Dynamics of multiple quantum coherences in dipole-coupling spins in solid. AB - A perturbation method deals with dipolar coupling spins in solids is presented. As example of application the method, the multiple-quantum coherence dynamics in clusters of a linear chain of four nuclear spins and a ring of six spins coupled by dipole-dipole interaction are considered. The calculated 0Q and 2Q intensities in a linear chain of four nuclear spins and 6Q intensity in a ring of six spins vs. the duration of the preparation period agree well with the exact solutions (for linear chain of four nuclear spins) and simulation data (for linear chain of four nuclear spins and a ring of six spin). PMID- 15885990 TI - Identifying the ERP correlate of a recognition memory search attempt. AB - Previous recognition memory studies have looked for differences in brain activity during recollection- and familiarity-based responding. Although an ERP component correlated with recollection success has been reported, no analogous component related to search initiation has been found. We argue that such a component has not been discovered because studies have compared trials in which participants have made a search attempt and failed (such as Know responses) with those in which the search attempt is successful (such as Remember responses). In the current study, we compared a task that required judgments of lifetime familiarity (differentiating famous from nonfamous names) with one that required judgments of episodic information (deciding whether a name was seen previously in the experiment). By comparing a task on which familiarity judgments were made with no search attempt to a second task in which a search attempt was likely to occur, we identified a component that may reflect the initiation of a memory search. This effect, maximal between 190 and 235 ms, is correlated with Old judgments in the episodic task. Previous ERP findings (e.g., FN400, parietal old/new effect) were also replicated in the present study. PMID- 15885991 TI - Magnetic field induced synthesis and self-assembly of super paramagnetic particles in a protein matrix. AB - Aqueous solution of a globular protein named bovine serum albumin was homogeneously mixed with ferrous and ferric ions and allowed to gel at ambient conditions. Gels were then oxidized using sodium hydroxide, in the presence of magnetic field of magnitude 0.13 T. The effect of magnetic field on the above biomimetic synthesis was a reduction in particle size and a directional assembly of synthesized super paramagnetic particles into a regular pattern in the protein film. The microstructural revelation was complimentary to Mossbauer results and magnetic measurement studies, i.e., an interesting variation in the magnetic behaviour of self-assembled super paramagnetic particles as a function of dc magnetic field induced ordering. PMID- 15885992 TI - Characterization of amphoteric multilayered thin films by means of zeta potential measurements. AB - Multilayer films of amphoteric methylated collagen were assembled on SOURCE 15S or SOURCE 15Q beads by sequential electrostatic deposition with negatively charged methylacrylic acid-hydroxyethyl methacrylate-methyl methacrylate (MAA HEMA-MMA) terpolymer. Methylated collagen and terpolymer were deposited under conditions where they were oppositely charged to one another, thereby facilitating growth of the films through electrostatic interactions. Measurements revealed alternating positive and negative zeta-potential with the deposition of each methylated collagen and terpolymer layer, respectively. Assembly pH had a remarkable influence on zeta-potential of the assembled multilayers and the deposition of methylated collagen will be frustrated when the assembly pH is up to 9.0. In addition, ionic strength (NaCl concentration) showed an intricate effect on zeta-potential of the films of amphoteric methylated collagen. PMID- 15885993 TI - [Prognosis of newborns of less than 26 weeks of gestation]. PMID- 15885994 TI - Impaired niacin sensitivity in acute first-episode but not in multi-episode schizophrenia. AB - Niacin (vitamin B3) flushing--a marker of altered prostaglandin signaling--is indirectly linked to the phospholipid-prostaglandin metabolism. Diminished skin flushing was repeatedly found in schizophrenia, but has not been systematically investigated at different stages of disorder as yet. We compared niacin sensitivity of 32 first-episode and 32 multi-episode patients (mainly on stable medication) with age and gender matched healthy controls. Methylnicotinate was applied in three concentrations onto the inner forearm skin. Flush response was assessed in 3 min intervals over 15 min using optical reflection spectroscopy. Whereas first-episode patients showed significantly diminished flush response as compared to controls, comparable differences were not found between multi-episode patients and controls. Comparison of niacin sensitivity at different stages of schizophrenia support the notion of altered prostaglandin signaling primarily at the onset of disorder. Longitudinal studies have to rule out possible long-term effects of neuroleptic medication. PMID- 15885995 TI - Exercise testing determines survival in patients with diffuse parenchymal lung disease evaluated for lung transplantation. AB - STUDY OBJECTIVES: Diffuse parenchymal lung disease is associated with a high risk of mortality despite early referral and listing for lung transplantation. We hypothesized that cardiopulmonary exercise test results and the distance walked in 6min (6MWTD) would be associated with survival in patients with diffuse parenchymal lung disease referred for lung transplantation. DESIGN: Retrospective cohort study. SETTING: Tertiary care center. PATIENTS: We included 51 consecutive patients with diffuse parenchymal lung disease who underwent exercise testing after referral to the Lung Transplant Program at the New York Presbyterian Hospital between January 2000 and December 2002. Thirty-three patients were listed, and 7 underwent transplantation during the study period. There were 17 deaths with 1 death post-transplantation. RESULTS: A 6MWTD < 350 m was associated with an increased risk of death (HR = 4.6, 95% CI 1.5-14.2, P = 0.009). Oxygen saturation with unloaded exercise (HR = 0.91, 95% CI 0.84-0.98, P = 0.015) and oxygen consumption at peak exercise adjusted for weight (HR = 0.88, 95% CI 0.79 0.99, P = 0.039) were also associated with the risk of death. A patient with oxygen saturation <95% during unloaded exercise had a 75% chance of dying on the list for transplantation. A patient with 6MWTD < 350 m had a 67% chance of dying on the list. CONCLUSIONS: Cardiopulmonary exercise test parameters and the 6MWTD were associated with the risk of death. Measures during exercise may be useful for determination of prognosis and for prioritizing patients with diffuse parenchymal lung disease for lung transplantation. PMID- 15885996 TI - Two different training programmes for patients with COPD: a randomised study with 1-year follow-up. AB - PURPOSE: To compare the effects on exercise capacity and health related quality of life (HRQoL) of two exercise programmes; one programme including endurance training and one including only resistance training and callisthenics. A second purpose was to find out whether the severity of chronic obstructive pulmonary disease (COPD) affected the training response and whether the interventions had a long-term effect. METHODS: Sixty-three patients were stratified according to severity of COPD and randomised to two training groups. Group A had a mixed programme including endurance training. Group B had resistance training and callisthenics. All trained twice weekly for 8 weeks. A symptom-limited ergometer test, 12-min walking test, dynamic spirometry, blood gas analysis at rest and HRQoL were measured before and after the training period. Follow-up tests were conducted at 6 and 12 months after training. RESULTS: Forty-two patients fulfilled the trial. In group A (n=20) peak exercise capacity increased by 7W (P<0.001) and 12-min walking distance (12MWD) by 50 m (P<0.01), whereas group B (n=22) did not change in any of these variables. HRQoL did not change significantly in either group. Training response was similar in patients with moderate and severe disease. One year post-training 12MWD had returned to pre training level in group A, and below pre-training level in group B (P<0.05). CONCLUSIONS: Exercise capacity in patients with severe and moderate COPD improved by intensive endurance training, two sessions a week for 8 weeks. The improvement was however small and HRQoL did not improve. Severity of illness did not affect response to training. The results indicated that the effects of a short endurance training intervention slowed down decline in baseline functional exercise capacity for 1 year. PMID- 15885997 TI - Exhaled nitric oxide as a screening tool for asthma in school children. AB - It is now widely accepted that augmented levels of fractional exhaled nitric oxide (FeNO) reflect airway inflammation and the methodology has been optimised for potential clinical use. We were interested in investigating whether this measurement can be used as a tool to screen and identify school children with asthma. To do this, FeNO was measured using an on-line single exhalation analyser in 368 children aged 8-10 years in six Oxfordshire primary schools, by two investigators blinded to the disease status of the children. The children were then categorised into 'normal', 'atopic asthma', 'non-atopic asthma' and 'atopy only' groups, according to their responses to the ISAAC questionnaire and perusal of the children's medical records kept by their family practitioners. Increased levels of FeNO were found in 'atopic asthmatic', 'non-atopic asthmatics' and 'atopic only' groups (median values of 24.4, 7.8 and 15.3 ppb, respectively, compared to normal controls' of 6.9 ppb). Levels were increased in atopic children regardless of whether they had asthma and were significantly higher than non-atopic asthmatics. We conclude that FeNO measurement is not a useful tool for identifying children with asthma in the community, as increased levels did not discriminate between those with asthmatic and atopic symptoms. PMID- 15885998 TI - Variability of nitric oxide metabolites in exhaled breath condensate. AB - The collection of exhaled breath condensate (EBC) is simple and non-invasive, however, there are few data on the methodological aspects affecting concentrations of compounds in EBC. The aim of this study was to investigate methodological issues for measuring nitric oxide metabolites (NO(x)) in EBC. Twenty-five healthy adults (12 females, age range 23-55 years) and 22 children (11 females, age range 7-6 years) were recruited for studies investigating inter- and intra-day repeatability, repeatability with controlled expiratory flows and temperature, flow dependence, and analytical variability of EBC NO(x). Both intra and inter-day repeatability was poor with a coefficient of repeatability of 103.4% of the mean difference between intra-day (15 min) measures and 118.6% of inter-day (24 h) differences. Repeatability was not improved when expiratory flow and temperature of the collection device were controlled. However, some of the variability (approximately 50%) may be accounted for by variability in the analytical technique (analytical variability) and this may result from difficulties in controlling for contamination. NO(x) levels were not affected by different expiratory flows in either adults or children but there was still significant variation within individuals. Levels of NO(x) in EBC seem to be highly variable and this needs to be considered if EBC NO(x) is to be used in clinical studies. PMID- 15885999 TI - Platinum nanoparticles-doped sol-gel/carbon nanotubes composite electrochemical sensors and biosensors. AB - Platinum nanoparticle-doped sol-gel solution is prepared and used as a binder for multi-walled carbon nanotubes (CNT) for the fabrication of electrochemical sensors. Amine group containing sol-gel solution is selected to utilize the affinity of -NH(2) groups toward metal nanoparticles for stabilization the nanoparticles in solution. The resulting CNT-silicate material brings new capabilities for electrochemical devices by using the synergistic action of the electrocatalytic activity of Pt nanoparticles and CNT. The combined electrocatalytic activity permits low-potential detection of hydrogen peroxide with remarkably improved sensitivity. With the incorporation of glucose oxidase within the Pt-CNT-silicate matrix, a Pt-CNT paste-based biosensor has been constructed that responds more sensitively to glucose than CNT-based biosensor. The influences of the composite of the sol-gel solution, the quantity of the solution and the Pt nanoparticles loading are examined. In pH 6.98 phosphate buffer, almost interference free determination of glucose is realized at 0.1 V versus SCE with a linear range from 1 to 25 mM, a response time <15s, and the sensitivity is 0.98 microA mM(-1)cm(-2). The sensitivity of the Pt-CNT paste based biosensor is almost four times larger than that of the CNT-based biosensor (0.27 microA mM(-1)cm(-2) at 0.1 V). The improved electrocatalytic activity and surface renewability made the Pt-CNT-silicate system a potential platform to immobilize different enzymes for other bioelectrochemical applications. PMID- 15886000 TI - Biosensor for rapid determination of 3-hydroxybutyrate using bi-enzyme system. AB - A bi-enzyme-based Clark electrode was developed for the determination of 3 hydroxybutyrate. This sensor is based on the specific dehydrogenation by 3 hydroxybutyrate dehydrogenase (HBDH, E.C. 1.1.1.30) in combination with salicylate hydroxylase (SHL E.C. 1.14.13.1). The enzymes were entrapped by a poly(carbamoyl) sulfonate (PCS) hydrogel on a Teflon membrane. The principle of the determination scheme is as follows: the specific detecting enzyme, HBDH, catalyses the specific dehydrogenation of 3-hydroxybutyrate consuming NAD(+). The products, NADH, initiate the irreversible decarboxylation and the hydroxylation of salicylate by SHL in the presence of oxygen. SHL forces the equilibrium of dehydrogenation of 3-hydroxybutyrate by HBDH to the product side by consuming NADH. Dissolved oxygen acts as an essential material for SHL during its enzymatic reactions. This results in a detectable signal due to the SHL-enzymatic consumptions of dissolved oxygen in the measurement of 3-hydroxybutyrate. Interferences from different amino acids and electroactive substances were found to be minimal due to the specificity of HBDH and the application of a Teflon membrane. The sensor has a fast response (2s) and short recovery time (2 min) with a linear range between 8 and 800 microM 3-hydroxybutyrate and a detection limit of 3.9 microM. A good agreement (R(2)=0.9925) with theoretical calculation was obtained in spiked serum sample measurements. PMID- 15886001 TI - Functional verification of pulse frequency modulation-based image sensor for retinal prosthesis by in vitro electrophysiological experiments using frog retina. AB - The functioning of a 16 x 16 pixel pulse frequency modulation (PFM) image sensor for retinal prosthesis is verified through in vitro electrophysiological experiments using detached frog retinas. This image sensor is a prototype for demonstrating the application to in vitro electrophysiological experiments. Each pixel of the image sensor consists of a pulse generator (PFM photosensor), a stimulus circuit, and a stimulus electrode (Al bonding pad). The image sensor is fabricated using standard 0.6 microm CMOS technology. For in vitro electrophysiological experiments, a Pt/Au stacked electrode is formed on the Al bonding pad of each pixel and the entire sensor is fixed in epoxy resin. The PFM image sensor is confirmed experimentally to provide electrical stimulus to the retinal cells in a detached frog retina. PMID- 15886002 TI - Amperometric biosensor based on tyrosinase-conjugated polysaccharide hybrid film: selective determination of nanomolar neurotransmitters metabolite of 3,4 dihydroxyphenylacetic acid (DOPAC) in biological fluid. AB - The amperometric detection of neurotransmitters metabolite of 3,4 dihydroxyphenylacetic acid (DOPAC) was achieved at a tyrosinase-chitosan composite film-modified glassy carbon (GC) electrode. The optimal conditions for the preparation of the biosensor were established. This bio-composite film was characterized by scanning electron microscopy (SEM) and Fourier transformed infrared (FT-IR) spectra, suggesting that chitosan covalently connected to chitosan chains. Electrochemical characterization of the bio-hybrid membrane covered electrodes were also performed in 0.05 M phosphate buffer solution (pH 6.52) containing neurotransmitters or their derivatives by using cyclic voltammetry (CV), linear sweep voltammetry (LSV), square wave voltammetry (SWV) and amperometry. This simply-prepared protein-polysaccharide hybrid film provides a microenvironment friendly for enzyme loading. The sensor was operated at -0.15 V with a short response time. The current linearly increased with the increasing concentration of DOPAC over the concentration of 6 nM-0.2 mM. The lower detection limit for DOPAC is 3 nM (S/N=3). The sensitivity of the sensor is 40 microA mM( 1). A physiological level of neurotransmitters and their derivatives including dopamine, l-dopa, adrenaline, noradrenaline and homovanillic acid as well as ascorbic acid, uric acid and acetaminophen do not affect the determination of DOPAC. PMID- 15886003 TI - A study exploring drug use and management of patients presenting to an inner city emergency department. AB - Patients who present to an emergency department (ED) with a problem related to illegal drug use can be difficult to identify and are perceived to generate a significant workload for staff. This study suggests that illicit drug use may be more common than previously reported and also that the impact on the ED is perceived by staff to be disproportionately high compared with the actual numbers of patients presenting with complaints related to illicit drug use. We conclude that the over estimation by staff is directly related to the challenges that staff working within the ED setting believe this group of patients pose. PMID- 15886004 TI - Discovery of a novel small molecule, 1-ethoxy-3-(3,4-methylenedioxyphenyl)-2 propanol, that induces apoptosis in A549 human lung cancer cells. AB - A novel small molecule, 1-ethoxy-3-(3,4-methylenedioxyphenyl)-2-propanol (EOD), was synthesized in our laboratory. Previously, we reported pharmacological properties of EOD, triggering apoptosis in Human umbilical vein endothelial cells (HUVECs). Here, we further investigated the effects of EOD on the growth of A549 human lung cancer cells. EOD treatment induced apoptosis in A549 cells via up regulating the expression of P53 protein, blocking cell cycle partly at G1 phase, and ultimately activating caspase-3. In contrast, caspase-8 might be irrelevant to EOD-triggered apoptosis. This study indicated that EOD might be a potential chemopreventive agent for lung cancer. The work would encourage us to add more novel compounds to our 'library' of small molecules derived through modern synthetic organic chemistry, and would drive us to determine the proteins that the compounds target. PMID- 15886005 TI - MCP-1 and murine prion disease: separation of early behavioural dysfunction from overt clinical disease. AB - Prion diseases are chronic, fatal neurodegenerative conditions of the CNS. We have investigated the role of monocyte chemoattractant protein-1 (MCP-1) in the ME7 model of murine prion disease. MCP-1 expression increased in the CNS throughout disease progression and was positively correlated with microglial activation. We subsequently compared the inflammatory response, pathology and behavioural changes in wild-type (wt) mice and MCP-1 knockout mice (MCP-1-/-) inoculated with ME7. Late-stage clinical signs were delayed by 4 weeks in MCP-1-/ mice, and survival time increased by 2-3 weeks. By contrast, early changes in affective behaviours and locomotor activity were not delayed in onset. There was also no difference in microglial activation or neuronal death in the hippocampus and thalamus of wt mice and MCP-1-/- mice. These results highlight an important dissociation between prolonged survival, early behavioural dysfunction and hippocampal/thalamic pathology when considering therapeutic intervention for human prion diseases and other chronic neurodegenerative conditions. PMID- 15886006 TI - An animal model of a spontaneously reversible obstructive sleep apnea syndrome in the monkey. AB - The anatomies of the tongue and uvula in monkeys share many similarities with humans, such that this species has the closest approximation to the human upper airway than any other species. In this study, we investigated the feasibility of using small monkeys as experimental animals for an obstructive sleep apnea model. Monkeys received intradermal injections of liquid collagen in the uvula, tongue, and lateral pharyngeal walls every 2 weeks. Polysomnography was performed bi monthly in order to control the impact of injections on breathing events, respiratory effort (as measured by esophageal pressure), and sleep. Before injections, the three animals showed normal breathing during sleep with a mean of 4.8 +/- 2.0 events/h. After injections, a mean of 27.9 +/- 19.7 hypopneas/h was recorded (P = 0.023). Total sleep time was significantly reduced, with a decrease of REM sleep and stage II sleep; however, stage I sleep increased. Collagen injections in monkey's upper airways can create sleep-disordered breathing and abnormal sleep, as seen in sleep apneic patients. PMID- 15886007 TI - DNA methylation disturbances as novel therapeutic target in lung cancer: preclinical and clinical results. AB - The prognosis of lung cancer is very much limited by the difficulties of diagnosing early stage disease amenable to surgery. Thus, novel diagnostic and therapeutic approaches are urgently needed for this common type of cancer. Recently, epigenetic alterations of tumor cells have been defined for a multitude of tissues and genes. Thus, promoter hypermethylation of tumor suppressor genes, and other targets of neoplasia-associated methylation disturbances, have become the most frequent recurrent alteration in solid tumors and hematologic neoplasia. In lung cancer, several sets of genes including the tumor suppressor gene p16, the DNA repair gene O(6)-methylguanine-DNA methyltransferase (MGMT), E-cadherin and retinoic acid receptor beta have been shown to be frequently methylated and inactivated. Distinct methylation patterns can provide molecular distinctions between different histologic subtypes of lung cancer. Gene hypermethylation in lung cancer is an early event associated with exposure to tobacco-specific carcinogens. Highly sensitive detection of hypermethylated DNA in sputum and peripheral blood offers a powerful tool for detecting lung cancer at an early stage. Epigenetic alterations in cancer, as opposed to genetic lesions, are potentially reversible. Thus, hypermethylation has been studied as a therapeutic target for agents which revert this epigenotype. The most advanced drugs to inhibit methylation are two azanucleosides, decitabine and its ribonucleoside analogue 5-azacytidine. In vitro, demethylating agents given at low doses reactivate tumor suppressor genes, and in mouse models, the development of lung cancer can be retarded. This effect is more powerful when histone acetylation, as a second epigenetic silencing mechanism, is also inhibited pharmacologically (HDAC inhibitors). Clinical trials of both groups of agents have been performed, and novel demethylating agents which are not incorporated into DNA offer further perspectives for epigenetic therapy of lung cancer and other malignancies. PMID- 15886008 TI - Coping and psychological well being in families of elderly cancer patients. AB - This article reviews research directions on elderly cancer patients and the consequences of their disease for their partners and families. In a systematic review of the research literature, five research directions were identified: first, the relationship between the family and the etiology of the disease; second, the importance of the family as a source of social support; third, family caregivers as second order patients and the moderating role of psychological factors; fourth, cancer as a challenge for the family and fifth, ways of providing social and psychological support for the families of patients including examples for intervention programs. It is shown that the assumption of a "psychogenic" etiology may inhibit adaptive processes. Depending on its quality, social support can trigger adaptive coping in the patients. Elderly patients seem to be ambivalent in their attitudes towards social support because they may feel hindered with respect to their autonomy. Distress among family members is sometimes very similar to the distress experienced by patients. Several studies have shown that distress is differentially influenced by disease-related, personal and interpersonal factors as well as social resources. Interpersonal factors seem to have a specific impact on the adaptive capacities of patients and their family members. Open communication, a "healthy" family cohesion and adaptive competence seem to positively influence patients' coping. Support programs which actively include family members seem to be effective in reducing the distress of patients and their relatives. Some conclusions will be drawn including a discussion of future research directions. PMID- 15886009 TI - Surgical interventions in patients with hematologic malignancies. AB - While surgery is not a part of treatment of hematological malignancies a significant number of patients suffering from such diseases need to undergo surgical interventions. These may comprise of very minor procedures such port implantation, however, already the indication for appendicectomy could be a difficult scenario being influenced by the underlying disease and side effects of the treatment. Often septic foci need to be cleared prior to start of systemic treatment. Out of the really major problems, the surgical resection of aspergillomas represent a demanding task accompanied by significant morbidity and mortality. Emergency situations sometimes are induced by treatment such as necrotizing enterocolitis but sometimes are mimicked by drug-induced abdominal symptoms. An experienced team of surgical and medical oncologists is required for successful management. As the number of long-term surviving patients increases, secondary problems as aseptic bone necrosis need to be addressed and adequately handled. Even at this stage it might be difficult to assess the patients risk for open-heart surgery adequately. This group of patients if scheduled for any operative intervention needs to undergo a very careful selection process and perfect perioperative management to make sure that the result of surgery are excellent and do not compromise further medical treatment of the underlying disease. PMID- 15886010 TI - Retinoids as chemoprevention for head and neck cancer: where do we go from here? AB - The retinoids, natural and synthetic derivatives of vitamin A, have a promising role in the chemoprevention of head and neck squamous cell carcinoma despite facing the limitations of toxicity as well as innate and acquired resistance. Adhering to the concept of field cancerization and following multi-step carcinogenesis, premalignant lesions of the head and neck have long been the focus of intervention with retinoids. Similarly, these agents have been applied towards preventing second primary malignancies from developing following curative therapy for upper aerodigestive cancers. Despite encouraging response rates, the need for overcoming innate and acquired retinoid resistance as well as minimizing retinoid-related toxicity will likely be relegated to the new generation of receptor-selective, synthetic retinoids. In the interim, retinoid-non-retinoid combinations as well as other classes of chemopreventive agents seem to provide promise in curtailing the incidence of aerodigestive malignancies. PMID- 15886011 TI - Interleukin-12 and interleukin-16 in periodontal disease. AB - The immune system plays an important role in the pathological process of periodontitis. Interleukin-12 (IL-12) is produced by monocytes, macrophages and neutrophils. These cells are proinflammatory infiltrates in periodontitis tissues. High IL-12 will contribute to the immune reaction to Th1 type. IL-12 is an inducer of INF-r production. IFN-gamma itself can also activate IL-12 production. Lipopolysaccharides (LPS) of periodontopathogens are also activators of IL-12. Interleukin-16 (IL-16) can cause the high affinity of IL-2 receptors on CD4+ cells and is chemotaxis to Th1 cells and CD4+ T cells. IL-16 can stimulate monocytes to produce proinflammatory cytokines and is highly associated with inflammation including arthritis, enteritis and allergic rhinitis. However, the information on IL-12 and IL-16 in periodontitis is not clear. In this study, 105 GCF samples were collected from 19 periodontal disease patients and 6 healthy ones. The clinical periodontal indices, the habits of cigarette smoking and alcohol drinking were recorded. ELISA was used to determine the levels of IL-12 and IL16 in the GCF. In the non-smoking/non-alcohol-drinking individuals: (1) the total amount of IL-12 (but not IL-16) was significantly higher in chronic periodontitis (CP) sites than gingivitis (G) or healthy (H) sites; (2) the diseased sites (CP + G) had a significantly higher total amount of IL-12 (but not IL-16) than the H sites. Among CP sites, both the concentration and total amount of IL-16 (but not IL-12) were significantly higher in alcohol drinkers/cigarette smokers as compared to the non-drinkers/non-smokers. CP sites of the drinkers/smokers also had significantly deeper probing pocket depth than sites of those without these two habits. IL-12 and IL-16 may be related to the pathogenesis of periodontal disease, but within the periodontitis sites, IL-16 may be related to disease severity in alcohol drinkers/smokers. PMID- 15886012 TI - Inhibition of Ca2+/calmodulin-dependent protein kinase II, RAS-GTPase and 20 hydroxyeicosatetraenoic acid attenuates the development of diabetes-induced vascular dysfunction in the rat carotid artery. AB - Diabetes causes accelerated vascular dysfunction through mechanisms that are poorly understood. This study examined the role of Ca2+/calmodulin-dependent protein kinase II (CaMKII), Ras-GTPase and 20-hydroxyeicosatetraenoic acid (20 HETE) in the development of abnormal reactivity to vasoactive agents in the carotid artery of diabetic rats. The vasoconstrictor response induced by endothelin-1 (ET-1) was significantly increased, whereas vasodilator response to carbachol was significantly reduced in the carotid artery segments of the STZ diabetic rats. In contrast, the vasoconstrictor response to depolarization of the carotid arterial rings with 50mM KCl was similar in control and diabetic animals. Chronic intraperitoneal administration of KN-93 (5 mg/kg/alt diem), an inhibitor of CaMKII, FPTIII (1.5 mg/kg/alt diem), an inhibitor of Ras-GTPase, and inhibitors of 20-HETE formation 1-aminobenzotriazole (ABT, 50 mg/kg/alt diem) and N-hydroxy-N'-(4-butyl-2-methylphenyl)formamidine (HET0016, 2.5mg/kg/day), produced significant normalization of the altered agonist-induced vasoconstrictor and vasodilator responses without affecting blood glucose levels. All the inhibitors were administered for 4 weeks starting from the day 1 of diabetes induction. Inhibition of CaMKII, Ras-GTPase or 20-HETE formation did not affect the agonist-induced vasoconstrictor and vasodilator responses in the non-diabetic control animals. These data indicate that chronic blockade of CaMKII, Ras-GTPase or the production of 20-HETE normalizes the altered vascular reactivity to ET-1 and carbachol in the carotid artery of STZ-induced diabetic rats. PMID- 15886013 TI - Arginine deprivation and metabolomics: important aspects of intermediary metabolism in relation to the differential sensitivity of normal and tumour cells. AB - Arginine deprivation causes many types of tumour cells to die, often because they cannot recover or convert urea cycle intermediates into arginine. The powerful homeostatic mechanisms that kicks in to restore arginine levels in vivo are lacking in vitro, where there is no supply of citrulline. Comparison between cells deprived of arginine by direct elimination methods or indirectly via arginine degrading enzymes should show differences depending on their ability to handle alternative intermediates (ornithine, citrulline and argininosuccinate) of the urea cycle. The internal state of cells that can, versus those that cannot, use intermediates will metabolically be quite different. These differences should provide clear indicators regarding the sensitivity (susceptibility) of cells to arginine deprivation, from which we will be in a much better position to judge which tumours to treat, and possibly how to design the best treatment to eliminate them. PMID- 15886014 TI - Disrupted compaction of CNS myelin in an OSP/Claudin-11 and PLP/DM20 double knockout mouse. AB - OSP/claudin-11 and PLP are both tetraspan proteins concentrated in CNS myelin. It has been proposed that they have a structural role in myelin formation and maintenance due to their localization and concentration in membrane sheaths. This hypothesis is not supported by the fact that both OSP/claudin-11- and PLP-null mice have relatively normal-appearing myelin and mild neurological deficits. Since both OSP/claudin-11 and PLP are abundant in myelin and have similar structures, the mild phenotypes of the knockout mice are likely due to compensatory mechanisms. Here we show that when both OSP/claudin-11 and PLP genes are knocked out, mice had severe neurological deficits, markedly abnormal myelin compaction, and smaller axon diameters. Interestingly, when either of these genes was knocked out, the expression of the other protein was increased. These data demonstrate that OSP/claudin-11 and PLP have essential structural functions in maintaining normal compact myelin and there is redundancy in their functions. PMID- 15886015 TI - Synaptotagmin mutants Y311N and K326/327A alter the calcium dependence of neurotransmission. AB - Synaptotagmin I, a calcium-binding synaptic vesicle protein, is thought to act as the calcium sensor for fast neurotransmission, but what synaptotagmin I does, upon binding calcium, to trigger exocytosis is still unknown. To begin to examine the role of synaptotagmin I's interactions with calcium-dependent binding partners, three mutant versions of synaptotagmin I reported to affect calcium dependent self-oligomerization (Y311N, K327A, and K326/327A) were expressed in cultured mouse hippocampal neurons lacking endogenous synaptotagmin I, and effects on neurotransmission were evaluated by comparison with transmission rescued by wild-type synaptotagmin I. All three mutants reduced transmitter release. To separate effects on calcium binding from effects on calcium-dependent oligomerization, we measured the calcium dependence of exocytosis for two of the mutants. Both showed apparent calcium affinity much lower than wild-type, a reduction sufficient to account for the neurotransmission defects. We conclude that self-oligomerization is unlikely to play any significant role in triggering synaptic vesicle exocytosis. PMID- 15886016 TI - GGA proteins regulate retrograde transport of BACE1 from endosomes to the trans Golgi network. AB - Golgi-localized, gamma ear-containing, ADP ribosylation factor-binding (GGA) proteins have been shown to be implicated in the sorting of cargo proteins from the trans-Golgi network (TGN) to endosomal compartments. GGAs directly bind to DXXLL motifs in the cytoplasmic domains of cargo proteins. The Alzheimer associated beta-secretase BACE1 also interacts with GGA proteins, but the functional relevance of this interaction was unknown. Here, we show that GGA1 regulates the retrograde transport of internalized BACE1 from endosomal compartments to the TGN by direct interaction in a phosphorylation-dependent manner. While phosphorylated BACE1 is efficiently transported from endosomes to the TGN, non-phosphorylated BACE1 enters a direct recycling route to the cell surface. Our data indicate that GGA proteins are not only involved in the sorting at the TGN but also mediate the retrograde transport of cargo proteins from endosomes to the TGN. PMID- 15886017 TI - Adenosine signaling promotes neuronal, catecholaminergic differentiation of primary neural crest cells and CNS-derived CAD cells. AB - In neural crest (NC) cultures cAMP signaling is an instructive signal in catecholaminergic, sympathoadrenal cell development. However, the extracellular signals activating the cAMP pathway during NC cell development have not been identified. We demonstrate that in avian NC cultures, evidenced by tyrosine hydroxylase expression and catecholamine biosynthesis, adenosine and not adrenergic signaling, together with BMP2, promotes sympathoadrenal cell development. In NC cultures, addition of the adenosine receptor agonist NECA in the presence of BMP2 promotes sympathoadrenal cell development, whereas the antagonist CGS 15943 or the adenosine degrading enzyme adenosine deaminase (ADA) suppresses TH expression. Importantly, NC cells express A2A and A2B receptors which couple with Gsalpha increasing intracellular cAMP. Employing the CNS derived catecholaminergic CAD cell line, we also demonstrate that neuronal differentiation mediated by serum withdrawal is further enhanced by treatment with IBMX, a cAMP-elevating agent, or the adenosine receptor agonist NECA, acting via cAMP. By contrast, the adenosine receptor antagonist CGS 15943 or the adenosine degrading enzyme ADA inhibits CAD cell neuronal differentiation mediated by serum withdrawal. These results support that adenosine is a physiological signal in neuronal differentiation of the CNS-derived catecholaminergic CAD cell line and suggest that adenosine signaling is involved in NC cell development in vivo. PMID- 15886018 TI - Expression and purification of the SsbB protein from Streptococcus pneumoniae. AB - The Gram positive bacterium, Streptococcus pneumoniae, has two genes, designated ssbA and ssbB, which are predicted to encode single-stranded DNA binding proteins (SSB proteins). We have shown previously that the SsbA protein is similar in size and in biochemical properties to the well-characterized SSB protein from Escherichia coli. The SsbB protein, in contrast, is a smaller protein and has no counterpart in E. coli. This report describes the development of an expression system and purification procedure for the SsbB protein. The ssbB gene was amplified from genomic S. pneumoniae DNA and cloned into the E. coli expression vector, pET21a. Although, we had shown previously that the SsbA protein is strongly expressed from pET21a in the E. coli strain BL21(DE3)pLysS, no expression of the SsbB protein was detected in these cells. However, the SsbB protein was strongly expressed from pET21a in the Rosetta(DE3)pLysS strain, a derivative of BL21(DE3)pLysS which supplies the tRNAs for six codons that are used infrequently in E. coli. The differential expression of the two SSB proteins in the parent BL21(DE3)pLysS strain was apparently due to the presence of two rare codons in the ssbB gene sequence that are not present in the ssbA sequence. Using the Rosetta(DE3)pLysS/pETssbB expression system, a protocol was developed in which the SsbB protein was purified to apparent homogeneity. DNA binding assays confirmed that the purified SsbB protein had single-stranded DNA binding activity. The expression and purification procedures reported here will facilitate further investigations into the biological role of the SsbB protein. PMID- 15886019 TI - Production and purification of recombinant human BLyS mutant from inclusion bodies. AB - B lymphocyte stimulator (BLyS), a member of the tumor necrosis factor superfamily, is an important regulator of B cell homeostasis. In BLyS-deficient mice, B cell development is severely perturbed. On the other hand, mice transgenic for BLyS developed autoimmune disorders, such as increased germinal center formation, production of autoantibodies, and Ig deposition in kidneys. The overexpression of BLyS was found in some human autoimmune diseases. These findings suggest that BLyS has a crucial role in the humoral immune response and may be a therapeutic target for some human autoimmune diseases. To construct and express the therapeutic vaccine BLyS, we coupled a foreign immunodominant T helper epitope to the N terminus of BLyS (named recombinant BLyS mutant, rBLySM) and expressed rBLySM in Escherichia coli. We have developed a purification process of rBLySM from inclusion bodies. A step-down urea concentration strategy was applied to the rBLySM renaturation process. By this strategy, a stable yield of 4.5mg purified rBLySM per gram of cell paste could be obtained. PMID- 15886020 TI - Neural correlates of high and craving during cocaine self-administration using BOLD fMRI. AB - Modern theories of drug dependence hold the hedonic effects of drug-taking central to understanding the motivation for compulsive drug use. Previous neuroimaging studies have begun to identify brain regions associated with acute drug effects after passive delivery. In this study, a more naturalistic model of cocaine self-administration (SA) was employed in order to identify those sites associated with drug-induced high and craving as measures of reward and motivation. Non-treatment seeking cocaine-dependent subjects chose both when and how often i.v. cocaine administration occurred within a medically supervised SA procedure. Both functional magnetic resonance imaging (fMRI) data and real-time behavioral ratings were acquired during the 1-h SA period. Drug-induced HIGH was found to correlate negatively with activity in limbic, paralimbic, and mesocortical regions including the nucleus accumbens (NAc), inferior frontal/orbitofrontal gyrus (OFC), and anterior cingulate (AC), while CRAVING correlated positively with activity in these regions. This study provides the first evidence in humans that changes in subjective state surrounding cocaine self-administration reflect neural activity of the endogenous reward system. PMID- 15886021 TI - Task-relevant modulation of primary somatosensory cortex suggests a prefrontal cortical sensory gating system. AB - Increasing evidence suggests that somatosensory information is modulated cortically for task-specific sensory inflow: Several studies report short-term adaptation of representational maps in primary somatosensory cortex (SI) due to attention or induced by task-related motor activity such as handwriting. Recently, it has been hypothesized that the frontal or prefrontal cortex may modulate SI. In order to test this hypothesis, we studied the functional organization of SI while subjects performed the Tower of Hanoi task. This task is known to be related to activation of frontal or prefrontal areas. The functional organization of SI while performing the Tower of Hanoi task was compared to the organization of SI during performing the same movements but without the Tower of Hanoi task and with rest. Topography of SI was assessed using neuromagnetic source imaging based on tactile stimulation of the first (D1) and fifth digits (D5). Performing the Tower of Hanoi task was accompanied by plastic changes in SI as indicated by significant shifts in the cortical representations of D1 and D5: They moved further apart during the Tower of Hanoi task compared to the control task containing the same movements but without the cognitive characteristic. Thus, we conclude that SI maps undergo dynamic modulation depending on motor tasks with different cognitive demands. The results suggest that this short-term plasticity may be regulated by a prefrontal-cortical sensory gating system. PMID- 15886022 TI - The effect of adult-acquired hippocampal damage on memory retrieval: an fMRI study. AB - Bilateral hippocampal pathology typically results in significant memory problems. Despite apparently similar structural damage, patients with such lesions can differ in the pattern of impairment and preservation of memory functions. Previously, an fMRI study of a developmental amnesic patient whose anoxic hippocampal damage was incurred perinatally revealed his residual hippocampal tissue to be active during memory retrieval. This hippocampal activity was apparent during the retrieval of personal and general facts relative to a control task. In this study, we used a similar fMRI paradigm to investigate whether residual hippocampal activation was present also in patient VC with adult acquired anoxic hippocampal pathology. VC's performance and reaction times on the experimental personal and general fact tasks were comparable to age-matched control subjects. However, in contrast to the elderly control sample and the previous developmental amnesic patient, his residual hippocampal tissue did not show activation changes during the experimental tasks. This finding indicates that patient VC's successful retrieval of personal and general facts was achieved without a significant hippocampal contribution. It further suggests that the hippocampal activation observed in the elderly controls and previous developmental amnesic patient was not necessary for successful task performance. The reason for this difference in hippocampal responsivity between VC and the developmental amnesic patient remains to be determined. We speculate that it may relate to the age at which hippocampal damage occurred reflecting plasticity within the developing brain, or to cognitive differences between VC, the developmental amnesic patient, and the control subjects. PMID- 15886023 TI - On-line automatic slice positioning for brain MR imaging. AB - In clinical brain MR imaging protocols, the technician collects a quick localizer and manually positions the subsequent scans using the localizer as a guide. We present a method for automatic slice positioning using a rapidly acquired 3D localizer. The localizer is automatically aligned to a statistical atlas representing 40 healthy subjects. The atlas contains the probability of a given tissue type occurring at a given location in atlas space and the conditional probability distribution of the multi-spectral MRI intensity values for a given tissue class. Accurate rigid alignment of each subject to an atlas ensures that all patients' scans are acquired in a consistent manner. A further benefit is that slices are positioned consistently over time, so that scans of patients returning for follow-up imaging can be compared side-by-side to accurately monitor the progression of illness. The procedure also helps ensure that left/right asymmetries reflect true anatomy rather than being the result of oblique slice positioning relative to the underlying anatomy. The use of an atlas based procedure eliminates the need to refer to a database of previously scanned images of the same patient and ensures corresponding alignment across scanners and sites, without requiring fiducial markers. Since the registration method is probabilistic, the registration error tends to increase smoothly in the presence of increasing noise and unusual anatomy or pathology rather than failing catastrophically. Translations and rotations relative to the atlas can be set so that planning can be done in anatomical space, rather than scanner coordinates, and stored as part of the protocol allowing standardization of slice orientations. PMID- 15886024 TI - Molecular phylogenetic studies of the Mylabrini blister beetles (Coleoptera, Meloidae). PMID- 15886025 TI - Insight into the phylogeny of African Clariidae (Teleostei, Siluriformes): implications for their body shape evolution, biogeography, and taxonomy. AB - The phylogenetic relationships among Clariidae species (Teleostei, Siluriformes) were assessed using 563 nucleotides of the cytochrome b mitochondrial gene. We included 32 Clariidae species representative of seven genera in our analysis. Hetropneustes fossilis (Heteropneustidae) and Clarotes laticeps (Claroteidae) were used as outgroups. The molecular data identified two evolutionary lineages that correspond on one hand to African species and on the other hand to Asian species. Morphological and osteological evolution in Clariidae did not follow an orthogenetic series. Species with robust body, strong ossified head, and large adipose fin were not ancestral ones and eel-like species were not phylogenetically related and represent independent adaptation to life in mud. Adaptation to life in deep water occurred two times independently in lake Tanganyika (with Dinotopterus cunningtoni) and in Lake Malawi (with Bathyclarias species). Molecular dating using a molecular clock of 1% divergence per million years and a comparison with fossils records allowed an estimate of the timing of cladogenesis within the species studied. The Clariidae family originated in Asia 50 MY ago but contemporary African and Asian studied species originated from a common ancestor that was present on the Arabian plate about 15 MY ago. Systematic implications of these results are also discussed. PMID- 15886026 TI - Correction of QT values to allow for increases in heart rate in conscious Beagle dogs in toxicology assessment. AB - INTRODUCTION: Estimation of a direct effect of drugs on the duration of the electrocardiogram (ECG) QT interval can be confused by drug-induced increases in heart rate (HR). The objective of this assessment was to identify a correction formula that adequately corrects QT over a wide range of HRs. METHODS: Paired recordings of HR and QT interval measurements were obtained from 177 conscious Beagle dogs from both sexes in 9 toxicology studies. ECGs used for this evaluation were collected in either control vehicle-treated dogs, or from dogs prior to the first dose of a daily dosing regimen. Where more than one recording was available per dog, only one was used in the analyses. The assessments were made based on the lowest and highest pre-dose HR for each dog. Correction factors according to [Bazett, H.C. (1920). An analysis of the time relationships or time relations of electrocardiograms. Heart 7:353-380], [Sagie, A., Larson, M.G., Goldberg, R.J., Bengtson, J.R., & Levy, D. (1992). An improved method for adjusting the QT interval for heart rate (the Framingham heart study). American Journal of Cardiology 70:797-801], [Fridericia, L.S. (1920). Die sytolendauer in elektrokardiogramm bei normalen menschen und bei herzkranken. Acta Medica Scandinavica 53:469-505.], [Todt, H., Krumpl, G., Krejcy, K. & Raberger, G. (1992). Mode of QT correction for heart rate: implications for the detection of inhomogeneous repolarization after myocardial infarction. American Heart Journal 124(3):602-609.] and [Van de Water, A., Verheyen, J., Xhonneux, R., & Reneman, R.S. (1989). An improved method to correct the QT interval of the electrocardiogram for changes in heart rate. Journal of Pharmacological Methods 22:207-217.] were applied to these QT intervals and plotted against HR. Linear regression statistical analyses using a single or multiple (i.e., baseline, sex and study) parameters model was then applied to trend lines. RESULTS: Although two correction factors ([Todt, H., Krumpl, G., Krejcy, K. & Raberger, G. (1992). Mode of QT correction for heart rate: implications for the detection of inhomogeneous repolarization after myocardial infarction. American Heart Journal 124(3):602-609.] and [Van de Water, A., Verheyen, J., Xhonneux, R., & Reneman, R.S. (1989). An improved method to correct the QT interval of the electrocardiogram for changes in heart rate. Journal of Pharmacological Methods 22:207-217.]) adequately corrected QT for changes in HR [i.e., slope of QTc versus HR not statistically significantly different from zero (p>0.05)] that of Van de Water showed a statistically superior correction. Although the method of analyses accounted for baseline, sex and study it was independently demonstrated that sex did not influence the outcome of the evaluations. Furthermore, higher HRs (i.e., maximum HRs for each dog) were better corrected than the lower HRs. In addition, statistical power analysis applied to these data showed that group sizes of 4-8 could, with 80% chance, detect a 10-5% change, respectively, in appropriately corrected QT. DISCUSSION: Overall, the data suggest that an evaluation of the most appropriate correction factor should be applied to each laboratory using their own data collected by their own method in their particular strain of dog. PMID- 15886028 TI - Potential role of centrioles in determining the morphogenetic status of animal somatic cells. AB - Irreversible differentiation (change of morphogenetic status) and programmed death (apoptosis) are observed only in somatic cells. Cell division is the only way by which the morphogenetic status of the offspring cells may be modified. It is known that there is a fixed limit to the number of possible cell divisions, the so-called 'Hayflick limit'. Existing links between cell division, differentiation and apoptosis make it possible to conclude that all these processes could be controlled by a single self-reproducing structure. Potential candidates for this replicable structure in a somatic cell are chromosomes, mitochondria (both contain DNA), and centrioles. Centrioles (diplosome) are the most likely unit that can fully regulate the processes of irreversible differentiation, determination and modification of the morphogenetic status. It may contain differently encoded RNA molecules stacked in a definite order. During mitosis, these RNA molecules are released one by one into the cytoplasm. In the presence of reverse transcriptase and endonuclease, RNA can be embedded in nuclear DNA. This process presumably changes the status of repressed and potentially active genes and, subsequently, the morphogenetic status of a cell. PMID- 15886027 TI - Factors associated with academic achievement in children with recent-onset seizures. AB - Children with chronic epilepsy are more at risk for achievement problems than either children without seizures or children with other chronic disorders. Factors that lead to such problems in children with epilepsy, however, are not well understood. Exploring these factors is important because academic underachievement can lead to poor social outcomes and contribute to underemployment or unemployment in adulthood. This descriptive, cross-sectional study investigated a group of children who had been diagnosed with seizures approximately 12 months previously, providing the opportunity to describe relationships among family and child characteristics; parent, child, and teacher responses; and child academic achievement at the same point in time across the sample. Seventy-two children had standardized test total battery scores, 101 had a teacher's rating of performance, and 67 had scores for both. Data were analyzed using multivariable regression. Child adaptive competency and seizure severity were associated with higher teacher ratings of academic performance (beta=0.73, p<0.0001 and beta=2.38, p=0.0182, respectively). Child adaptive competency was associated with higher total battery scores (beta=0.73, p<0.0001). Contrary to findings in studies of children with chronic epilepsy, mean academic achievement in this sample of children with recent-onset seizures was in the average range; however, 25% of the children were at or below one standard deviation below the mean on the teacher's rating of performance and 10% on the total battery. It is therefore important for health professionals and educators to regularly assess the child's academic functioning and school performance to identify those at risk for problems. Health professionals and educators need to collaborate on assessment and interventions to help maximize child academic success. PMID- 15886029 TI - Acute airway obstruction complicating unilateral carotid endarterectomy. PMID- 15886030 TI - Transcatheter embolisation of visceral pseudoaneurysms--technical difficulties and modification of embolisation technique. AB - We report 13 successful emergency embolisations for 12 pseudoaneurysms performed in 10 patients. For five pseudoaneurysms the embolisation technique was modified according to number of supplying vessels, flow rate of pseudoaneurysms, vascular anatomy and whether there was clinical evidence of re-bleeding or not. Apart from traditional embolisation technique, modified embolisation techniques are also useful for endovascular therapy of pseudoaneurysms. PMID- 15886031 TI - Gradient-enhanced FAWSETS perfusion measurements. AB - This work describes the use of custom-built gradients to enhance skeletal muscle perfusion measurements acquired with a previously described arterial spin labeling technique known as FAWSETS (flow-driven arterial water stimulation with elimination of tissue signal). Custom-built gradients provide active control of the static magnetic field gradient on which FAWSETS relies for labeling. This allows selective, 180 degrees modulations of the phase of the perfusion component of the signal. Phase cycling can then be implemented to eliminate all extraneous components leaving a signal that exclusively reflects capillary-level perfusion. Gradient-enhancement substantially reduces acquisition time and eliminates the need to acquire an ischemic signal to quantify perfusion. This removes critical obstacles to application of FAWSETS in organs other than skeletal muscle and makes the measurements more desirable for clinical environments. The basic physical principles of gradient-enhancement are demonstrated in flow phantom experiments and in vivo utility is demonstrated in rat hind limb during stimulated exercise. PMID- 15886032 TI - Photo-CIDNP NMR spectroscopy of a heme-containing protein. AB - There are relatively few examples of the application of photo-CIDNP NMR spectroscopy to chromophore-containing proteins. The most likely reason for this is that simultaneous absorption of light by the photosensitiser molecule and the protein chromophore reduces the effectiveness of the photochemical reaction that produces the observed nuclear polarisation. We present details of experiments performed on the air-oxidised form of a small cytochrome, from the thermophilic bacterium Hydrogenobacter thermophilus, using both the wild-type protein and apo and holo forms of a double alanine b-type mutant. We show that, along with the apo state, it is possible to generate CIDNP in the air-oxidised form of the b type mutant, but not in the corresponding c-type cytochrome. This finding is supported by control experiments using horse-heart cytochrome c. PMID- 15886033 TI - Simulating adsorption of n-heptane in the Pt/Al2O3 model: influence of platinum. AB - The molecular modeling method refered to in the literature as Grand Canonical Monte Carlo was used to analyze the phenomenon of n-heptane adsorption on the Pt/gamma-Al(2)O(3) catalyst. In order to describe relevant interactions, use was made of the forcefield methods (UFF and CVFF). With the conditions adopted for the purpose of the study, Pt was found to exert an advantageous effect on the adsorption of n-heptane. The number of adsorbed molecules was related to the content of the noble metal, and the relation was directly proportional, when temperature and pressure were constant. The contribution of Pt was most distinct at 573 K and 100 kPa. PMID- 15886034 TI - Elucidation of the Na+, K+-ATPase digitalis binding site. AB - Despite controversy over their use and the potential for toxic side effects, cardiac glycosides have remained an important clinical component for the treatment for congestive heart failure (CHF) and supraventricular arrhythmias since the effects of Digitalis purpurea were first described in 1785. While there is a wealth of information available with regard to the effects of these drugs on their pharmacological receptor, the Na(+), K(+)-ATPase, the exact molecular mechanism of digitalis binding and inhibition of the enzyme has remained elusive. In particular, the absence of structural knowledge about Na(+), K(+)-ATPase has thwarted the development of improved therapeutic agents with larger therapeutic indices via rational drug design approaches. Here, we propose a binding mode for digoxin and several analogues to the Na(+), K(+)-ATPase. A 3D-structural model of the extracellular loop regions of the catalytic alpha1-subunit of the digitalis sensitive sheep Na(+), K(+)-ATPase was constructed from the crystal structure of an E(1)Ca(2+) conformation of the SERCA1a and a consensus orientation for digitalis binding was inferred from the in silico docking of a series of steroid based cardiotonic compounds. Analyses of species-specific enzyme affinities for ouabain were also used to validate the model and, for the first time, propose a detailed model of the digitalis binding site. PMID- 15886035 TI - A comparison of the counteracting effects of glycine betaine and TMAO on the activity of RNase A in aqueous urea solution. AB - Trimethylamine-N-oxide (TMAO) and glycine betaine are counteracting osmolytes found in cellular systems under osmotic stress, often in association with high urea concentrations. TMAO is a characteristic component of cartilaginous fish and marine molluscs, while glycine betaine is more widely distributed, occurring in plants, bacteria and the mammalian kidney. As part of a project to explain and understand the action of these methylamines, the RNase A-catalysed degradation of polyuridylic acid in the presence of urea and various osmolytes (0-1.0 M) was studied using (31)P Nuclear Magnetic Resonance spectroscopy. The decrease in reaction rate induced by urea could be fully recovered with 1 molar equivalent of trimethylamine-N-oxide or 1.4 molar equivalents of glycine betaine. These results indicate that the modification of RNase A activity induced by urea is not associated with gross irreversible structural changes and that both glycine betaine and trimethylamine-N-oxide have kinetically detectable counteracting effects. PMID- 15886036 TI - The significance of the activity of dissolved oxygen, and other gases, enhanced by high hydrostatic pressure. AB - The partial pressure of oxygen and other gases dissolved in water and subjected to high hydrostatic pressure is increased. Although this was established many years ago it remains a problematical phenomenon. The review deals with some of the underlying theoretical difficulties and discusses the kinetic and environmental implications of the pressure-enhanced partial pressures. PMID- 15886037 TI - Amphioxus allantoicase: molecular cloning, expression and enzymatic activity. AB - Allantoicase, one of the purine metabolism enzymes, is progressively truncated during the chordate evolution, yet it is unknown when its activity became phylogenetically extinct. In this study, a cDNA encoding allantoicase was isolated from the gut cDNA library of amphioxus Branchiostoma belcheri tsingtauense. It is 2441 bp long, and contains an open reading frame encoding a protein of 392 amino acid residues. RT-PCR analysis showed that amphioxus allantoicase was strongly expressed in the hepatic caecum, and weakly expressed in other tissues including hind-gut, gill, muscle, notochord, testis and ovary. The parallel experiment was performed measuring the allantoicase activity in the same tissues revealed that its activity was high in the hepatic caecum, but low or undetectable in other tissues examined. These suggest that allantoicase remains in action in the primitive chordate amphioxus. PMID- 15886038 TI - Embryonic development within carotenoid-enriched eggs influences the post-hatch carotenoid status of the chicken. AB - Carotenoids in the diet of the laying hen are incorporated into the egg yolk and subsequently into the liver and other tissues of the chicken embryo. Since these pigments are known to provide a range of health benefits to a variety of animals, it is of interest to know whether the effects of maternally derived carotenoids are strictly limited to the embryonic period or if they persist in the progeny after hatching. The aim of this study is to compare the effectiveness of pre hatch (from the hen's diet) with that of post-hatch (from the progeny's diet) supplementation with carotenoids on the carotenoid status of the chick during the first 4 weeks of post-hatch life. Hens were fed a control diet or a diet supplemented with a carotenoid-rich extract of alfalfa. Eggs from the supplemented hens contained up to 22 times more carotenoids than the controls. The concentration of carotenoids in the livers of chicks hatching from the enriched eggs was initially 29 times greater than in the control chicks. Hepatic carotenoid concentrations in chicks from enriched eggs maintained post-hatch on the control diet were sustained at higher values compared with chicks from control eggs that were fed post-hatch on the carotenoid-supplemented diet, for at least the first 7 days. However, by 14 days, the latter group had overtaken the former in terms of liver carotenoid levels. Thus, under these conditions, maternal effects predominate for at least the first week after hatching, whereas from 2 weeks onwards, the progeny's diet becomes the main determinant of its carotenoid status. Since the antioxidant and immunostimulatory roles of carotenoids are likely to be especially important during the immediate post-hatch period, maternal dietary intake of carotenoids may have important ramifications for the viability of the offspring. PMID- 15886039 TI - The isolation, characterization, and expression of a novel GDF11 gene and a second myostatin form in zebrafish, Danio rerio. AB - In the current study, the first non-mammalian growth/differentiation factor (GDF) 11-like homolog was cloned from zebrafish. At the nucleotide level, zebrafish GDF11 is most similar to human GDF11 (79%), while the peptide is most similar to mouse GDF11 (78%). Phylogenetic analysis showed that the zebrafish GDF11 clusters with mammalian GDF11s. This study also cloned a second MSTN form in zebrafish most similar to Salmonid MSTN2 forms. Based on real time PCR, GDF11 is expressed in multiple adult tissues, with levels highest in whole heads and gonads, and expression is less ubiquitous when compared to MSTN expression. During embryonic development, real time PCR demonstrated increasing GDF11 mRNA levels 10 h post fertilization (hpf), while MSTN mRNA levels remain low until 48 hpf. This is the first report of a transforming growth factor (TGF)-beta superfamily member in a non-mammalian species that is more closely related to GDF11 than MSTN, and also a second form of MSTN in zebrafish; suggesting that a more complex TGF-beta superfamily array exists in primitive vertebrates than previously thought. PMID- 15886040 TI - Glucose tetrasaccharide as a biomarker for monitoring the therapeutic response to enzyme replacement therapy for Pompe disease. AB - A tetraglucose oligomer, Glcalpha1-6Glcalpha1-4Glcalpha1-4Glc, designated Glc4, has been shown to be a putative biomarker for the diagnosis of Pompe disease. The purpose of this study was to assess whether Glc4 could be used to monitor the therapeutic response to recombinant human acid alpha glucosidase (rhGAA) enzyme replacement therapy (ERT) in patients with Pompe disease. Urinary Glc4 levels in 11 patients receiving rhGAA therapy was determined by both HPLC-UV and stable isotope dilution ESI-MS/MS. Combined Glc4 and maltotetraose, Glcalpha1-4Glcalpha1 4Glcalpha1-4Glc, (M4) concentrations, designated Hex4, in plasma from these patients were measured by HPLC-UV only. Baseline urinary Glc4 and plasma Hex4 in these patients (mean+/-SD: 34.2+/-11.3 mmol/mol creatinine and 1.7+/-0.8 microM, respectively) were higher than age-matched control values (mean+/-SD, 6.1+/-5.1 mmol/mol creatinine and 0.22+/-0.15 microM, respectively). Both urinary Glc4 and plasma Hex4 levels decreased after initiation of ERT for all patients. In the four patients with the best overall clinical response in both skeletal and cardiac muscle, levels decreased to within, or near, normal levels during the first year of treatment. In contrast, levels fluctuated and were persistently elevated above the control ranges in those patients with a less favorable clinical response (good cardiac response but limited motor improvement). These results suggest that urinary Glc4 and plasma Hex4 could serve as a valuable adjunct to clinical endpoints for monitoring the efficacy of therapeutic interventions such as rhGAA ERT in Pompe disease. PMID- 15886041 TI - Mitochondrial voltage-dependent anion channel gene family in Drosophila melanogaster: complex patterns of evolution, genomic organization, and developmental expression. AB - Voltage-dependent anion channels (VDACs), also known as mitochondrial porins, are a family of small pore-forming proteins of the mitochondrial outer membrane found in all eukaryotes. VDACs play important roles in the regulated flux of metabolites between the cytosolic and mitochondrial compartments, energy metabolism, and apoptosis. Annotation of the genome sequence of Drosophila melanogaster revealed three genes (CG17137, CG31722-A, and CG31722-B) with homology to porin, the previously described Drosophila VDAC. Molecular analysis reveals a complex pattern of organization and expression. The genomic organization of these four genes and sequence comparisons with other insect VDAC homologs indicate that this gene family evolved through a mechanism of duplication and divergence from an ancestral VDAC gene during the radiation of the genus Drosophila. CG17137, CG31722-A, and CG31722-B are expressed in a male specific pattern on both transcriptional and translational levels, while porin is equally expressed in both male and female flies. Additionally, CG31722-A and CG31722-B are expressed as a dicistronic transcript. Western blot analysis and immunofluorescence microscopy confirm that these proteins localize to the mitochondrion. Further expression analysis showed that CG17137 and CG31722-B are abundant in testes, while porin is ubiquitously expressed. While porin, CG17137, and CG31722-B are expressed to different degrees during embryogenesis, all of these proteins are dramatically reduced relative to cytochrome c content during larvogenesis. These studies illustrate a complex genomic organization and spatiotemporal pattern of expression for Drosophila VDACs as well as an evolutionary history consistent with either a partitioning of VDAC functions or an acquisition of novel functions among isoforms. PMID- 15886042 TI - Is the functional decline of Parkinson's disease similar to the functional decline of Alzheimer's disease? AB - Since many Parkinson's disease (PD) subjects develop dementia, we determined whether the correlation between functional and cognitive decline seen in Alzheimer's disease (AD) is seen in PD. Seventy-five PD subjects with and without dementia and 103 AD/MCI subjects underwent the Functional Assessment Staging (FAST), the Global Deterioration Scale (GDS), the UPDRS motor portion, and the MMSE. In AD/MCI subjects, changes in FAST and GDS scores correlated with MMSE (rho=-0.814, P<0.001; rho=-0.840, P<0.001, respectively). In PD subjects, the FAST and GDS also correlated with MMSE (rho=-0.675, P<0.001; rho=-0.647, P<0.001, respectively). The UPDRS correlated with the GDS and FAST more closely in PD than in AD. Similar to AD, functional declines in PD correlates with cognitive decline and may be influenced by motor disability in PD. PMID- 15886043 TI - Psychometric characteristics of the Parkinson's disease questionnaire (PDQ-39)- Ecuadorian version. AB - This study sought to analyse certain metric characteristics of the Ecuadorian version (EV) of the Parkinson's Disease Questionnaire (PDQ-39 EV). A cross sectional study was conducted on 137 Parkinson's disease (PD) patients attending a Movement Disorders Unit. Neurologists' assessments were based on Hoehn and Yahr (HY), Schwab and England and Unified Parkinson's Disease Rating Scales. Patients' self-evaluations included the Hospital Anxiety and Depression Scale, the Parkinson's Disease Quality of Life questionnaire (PDQL EV), and the PDQ-39 EV. Analyses for acceptability, internal consistency, precision, and construct validity (convergent and known-groups) were performed. Distribution of scores was satisfactory. There was no evidence of floor or ceiling effects. Although the alpha coefficient exceeded 0.70 for mobility, activities of daily living (ADL) and stigma, it was nevertheless low for bodily discomfort (0.48), communication (0.40), and social support (0.33). Fourteen items yielded low correlation coefficients (<0.40) with their respective dimensions. Correlation of social support and bodily discomfort with the Summary Index (SI) was modest (0.46 and 0.36, respectively). PDQ-39 EV SI convergent validity with the PDQL EV SI was very high (r(S)=-0.91), and known-groups validity proved satisfactory. Results agreed in part with those yielded by an international study, identifying specific flaws probably linked to socio-cultural influence. PMID- 15886044 TI - Progression and bilateralisation of motor signs in Parkinson's disease. PMID- 15886045 TI - Flexor carpi radialis motoneuron pool in subjects with chronic carpal tunnel syndrome are more excitable than matched control subjects. AB - We compared excitability of the flexor carpi radialis (FCR) motoneuron pool in subjects with and without carpal tunnel syndrome (CTS). The study involved 11 subjects with chronic idiopathic CTS and 11 asymptomatic subjects as controls. The H-reflex and M-response of FCR muscle were obtained by stimulating the median nerve in the cubital fossa in the presence of an isometric background contraction using surface stimulating and recording electrodes. There was a significantly higher H-reflex latency and amplitude and Hmax/Mmax in CTS subjects (P<0.05). Latency and amplitude of the M-response remained unaffected in CTS group. The results support the hypothesis that central hypersensitivity does occur in chronic CTS. Therefore even in the presence of pathology in peripheral structures central mechanisms should be considered by clinicians. PMID- 15886046 TI - Monitoring the change: current trends in outcome measure usage in physiotherapy. AB - Physiotherapists have traditionally relied on impairment measures such as range of motion and muscle strength to monitor patient progress. The impact of treatment on patients' daily activities can be assessed with valid and reliable questionnaires, but the use of standardized questionnaires by physiotherapists appeared to be limited. A range of strategies were implemented that aimed to increase physiotherapists' use of standardized measures of functional activities. A simple random sample of 300 was drawn from a database of physiotherapy providers to a transport accident scheme, and was surveyed in March and September 2003, with response rates of 51% and 55%, respectively. There was a statistically significant (P<.05) increase in reported use of seven questionnaires and a significant reduction in the perception of barriers that were targeted by the interventions The most frequently utilized tests were a pain rating scale and questionnaires for lumbar and cervical problems. Physiotherapists' attitudes to outcome measurement were generally positive although there was a small but statistically significant (P=.02) reduction in mean attitude score over the re test period. Physiotherapists in the population sampled significantly increased their reported use of a range of standardized outcome measures over the re-test period. The trend towards greater objectivity in measuring the progress of rehabilitation can enable physiotherapists to develop improved treatment plans with the patients' needs at the centre of the equation. PMID- 15886047 TI - The osteoblast-heparan sulfate axis: control of the bone cell lineage. AB - During osteogenesis, mesenchymal stem cells are recruited to the osteoblast lineage and progressively differentiate into osteoblasts that produce a mineralised extracellular matrix. Although most of the organic component of this matrix is comprised of collagen, growing evidence suggests the most bioactive element of a developing matrix is its heparan sulfate glycosaminoglycan complement. This species of linear, unbranched sugars contain protein-binding domains that regulate the flow of an astonishing number of mitogenic influences that coordinate mesenchymal stem cell commitment and growth, and ultimately, osteoblast phenotype. Among the heparan sulfate-binding factors known to be important to this process are sonic hedgehog, the fibroblast growth factors and their receptors, members of the transforming growth factor superfamily, as well as the collagens, laminins and fibronectins. How these sugars change during development to bring together the right combination of mitogenic/differentiative influences to trigger the successive phases of osteogenesis is currently the focus of intense research. PMID- 15886048 TI - The role of the growth hormone/insulin-like growth factor axis in tumor growth and progression: Lessons from animal models. AB - Over the past two decades it has become widely appreciated that a relationship exists between the insulin-like growth factors (IGFs) and cancer. Many cancers have been shown to overexpress the IGF-I receptor and produce the ligands (IGF-I or IGF-II) and some combinations of the six IGF-binding proteins. With the recent demonstration by epidemiological studies that an elevated serum IGF-I level is associated with an increased relative risk of developing a number of epithelial cancers, interest has been sparked in this area of research with the possibility of targeting the IGF-I receptor in cancer treatment protocols. This review highlights many of the most relevant studies in this exciting area of research, focusing in particular on lessons learned from animal models of cancer. PMID- 15886049 TI - Regulation of growth factor signaling and cell cycle progression by cell adhesion and adhesion-dependent changes in cellular tension. AB - The proliferation of most non-transformed cell types requires cell adhesion and cellular tension as well as exposure to mitogenic growth factors. Integrins and cadherins provide the adhesion signals, which ultimately allow for the cytoskeletal changes that control cellular tension. This review discusses the roles of integrins, cadherins, and the actin cytoskeleton as mediators of the mechanical tension critical for growth factor-dependent signaling and cell cycle progression. PMID- 15886050 TI - Protective effect of apigenin on radiation-induced chromosomal damage in human lymphocytes. AB - The potential use of flavonoids as a radioprotector is of increasing interest because of their high antioxidant activity and abundance in the diet. The aim of this study is to examine genotoxic and radioprotective effects of one of the most common flavonoids, apigenin, on radiation-induced chromosome aberrations in human lymphocytes. The cytokinesis-block micronucleus (CBMN) assay was used to evaluate such effects of apigenin. Blood samples were collected from two non-smoking healthy male volunteers who had no history of previous exposure to other clastogenic agents. Isolated lymphocytes were cultured. There were two tubes per concentration for all treatments. To evaluate the genotoxicity of apigenin, cells were first treated with different concentrations of apigenin (0, 2.5, 5, 10 and 25 microg/mL) at 24 h after culture initiation, followed by cytochalasin-B (Cyt B) treatment (3 microg/mL) and cell harvest at 44 and 72 h, respectively. Secondly, to investigate the radioprotective effect, cell cultures were exposed to different concentrations of apigenin as described above for 30 min before being irradiated to 2 Gy of 137Cs gamma rays (at a dose rate of 0.75 Gy/min). In all instances, the frequency of MN was scored in binucleated (BN) cells. The nuclear proliferation index also was calculated. We did not detect an increase in the frequency of MN in non-irradiated human lymphocyte cultures treated with 2.5, 5.0 or 10 microg/mL apigenin; although, we did observe an increase in cultures treated with 25 microg/mL apigenin (the highest concentration of apigenin used in our study). We also observed a significant increase in the frequency of MN in irradiated cells overall; however, the frequency was decreased as the concentration of apigenin increased, suggesting a radioprotective effect. These findings provide a basis for additional studies to help clarify the potential use and benefit of apigenin as a radioprotector. PMID- 15886051 TI - Orthovanadate increased the frequency of aneuploid mouse sperm without micronucleus induction in mouse bone marrow erythrocytes at the same dose level. AB - The objective of the current study was to investigate the ability of orthovanadate to induce aneuploidy in mouse sperm and micronuclei in mouse bone marrow cells at the same dose levels. The BrdU-incorporation assay was performed to test if the chemical treatment altered the duration of the meiotic divisions. It was found that orthovanadate (25mg/kg bw) treatment did not cause meiotic delay. To determine the frequencies of hyperhaploid and diploid sperm, male mice were treated by intraperitoneal (i.p.) injection with 5, 15 or 25mg/kg bw orthovanadate and sperm were sampled from the Caudae epididymes 22 days later. Fluorescence in situ hybridization (FISH) was performed with DNA-probes for chromosomes 8, X or Y. Significant increases in the frequencies of total hyperhaploid sperm (p<0.01) were found with 15 and 25mg/kg bw orthovanadate, indicating induced non-disjunction during male meiosis. The dose-response was described best by a linear equation. Orthovanadate did not significantly increase the frequencies of diploid sperm at any of the three doses tested, indicating that no complete meiotic arrest occurred. Orthovanadate was investigated also by the micronucleus test at i.p. doses of 1, 5, 15 or 25mg/kg bw, followed by bone marrow sampling 24h after treatment. None of the orthovanadate doses caused a significant increase in the rates of micronuclei (MN). Since the results show that orthovanadate induced non-disjunction during male meiosis without an accompanying induction of MN in bone marrow erythrocytes under the present experimental conditions and doses, it is concluded that male germ cells (meiosis) are more sensitive to the aneugenic effects of orthovanadate than somatic cells (mitosis). However, induction of micronuclei was reported in the literature with orthovanadate, vanadylsulfate and ammonium metavanadate, which contradicts the notion that vanadium compounds might be unique germ cell aneugens. PMID- 15886052 TI - Lack of mutagenicity of chromium picolinate in the hypoxanthine phosphoribosyltransferase gene mutation assay in Chinese hamster ovary cells. AB - Chromium picolinate (CrPic, Chromax) is a dietary supplement that is stable and more bioavailable than other commercially available forms of chromium. Chromium supplementation is known to enhance the action of insulin, particularly in insulin resistance and type 2 diabetes mellitus. A previous study reported that CrPic produced increases in mutations of the hypoxanthine phosphoribosyltransferase (Hprt) gene in Chinese hamster ovary (CHO) cell mutation tests. This study, however, evaluated CrPic produced by the testing laboratory and used an atypical 48 h exposure period for this test system. The current study evaluated the mutagenic potential of the most widely utilized commercial form of CrPic in CHO/Hprt mutation tests following International Conference on Harmonisation (ICH) Guidelines (+/-S9 metabolic activation with a 5h exposure) in addition to repeating the test with a 48 h exposure period -S9 activation. CrPic was suspended in dimethyl sulfoxide (DMSO) up to a concentration of 50 mg/mL; exposures were conducted under conditions in which precipitate was not evident and under conditions in which some precipitate of CrPic was visually evident in the cell culture medium at the highest concentrations (500 microg/mL). The concentrations evaluated for mutagenicity ranged from 15.6 to 500 microg/mL (+S9 and -S9) for the 5 h exposure and 31.3-500 microg/mL for the 48 h exposure (-S9). Only a slight degree of cytotoxicity was seen in the standard tests up to the limit of solubility in the medium. Toxicity, i.e., cloning efficiency < or =50% of the solvent control, but no mutagenic increases were observed at 500 microg/mL following a 48 h exposure period. The results of these studies showed that CrPic was non-mutagenic in two independent CHO/Hprt assays and in an assay using a 48 h exposure period. PMID- 15886053 TI - Withdrawal syndrome from gamma-hydroxybutyric acid (GHB) and 1,4-butanediol (1,4 BD) in Sardinian alcohol-preferring rats. AB - Gamma-hydroxybutyric acid (GHB) and its precursors, 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL), are recreational drugs widely abused in the US, Europe and Australasia. A severe withdrawal syndrome from GHB, 1,4-BD and GBL has been increasingly documented over the last years, necessitating the development of a reliable animal model for investigations of potential therapeutic approaches. The present study describes the induction and occurrence of audiogenic seizures as a sign of withdrawal from GHB and 1,4-BD in selectively bred Sardinian alcohol preferring (sP) rats, treated with escalating doses of GHB (1.5-3.5 g/kg, twice daily; i.g.) or 1,4-BD (500-1000 mg/kg, twice daily; i.g.) for 9 consecutive days. Acute administration of the selective GABA(B) receptor antagonist, SCH 50911, dramatically increased seizure occurrence. We propose that the inherent sensitivity of sP rats to different GHB-associated responses may have contributed to the unraveling of a phenomenon which was otherwise not recognizable in other rat strains. PMID- 15886054 TI - Hepatoprotection against oxidative damage. PMID- 15886055 TI - Mild conditions for releasing mono and bis-biotinylated macromolecules from immobilized streptavidin. AB - The high affinity (kd= approximately 10(-15)M) of streptavidin and avidin for biotin is key to a large number of biological applications and is essentially irreversible unless the complex is exposed to harsh conditions (e.g. heat (100 degrees C for 10 min)), detergents, and/or denaturants which damage macromolecules. Thus, high binding affinity becomes a disadvantage when a biotinylated target must be released for further processing. This work describes relatively mild conditions that release biotin and mono- and bis-biotinylated macromolecules from immobilized streptavidin on monodispersed magnetic beads. PMID- 15886056 TI - Fragility--the price of renewed life. Patients experiences of open heart surgery. AB - BACKGROUND: Open heart surgery often implies a threat to life and is associated with fear and anxiety. It is also a strong encroachment on body and integrity and adjusting life afterwards could be difficult. Despite improvements in treatment the patients' reactions appear to be unchanged. Introducing a lifeworld perspective would supply a different kind of knowledge based upon the patients' own experiences coloured by their linguistic usage and bodily expressions. AIM: The aim of this study was to describe patients' experiences of open heart surgery in a lifeworld perspective. METHOD: Fourteen patients treated with coronary artery bypass surgery and/or heart valve operation were in-depth interviewed in 2003. The phenomenological method was used for the interviews as well as for the analysis. The informants reflected on their experiences of the illness, meetings with health care, family relations and wishes for the future. FINDINGS: The essence of the phenomenon was fragility. Fragility was understood through the following categories: distance, uncertainty, vulnerability, reliance and gratitude. CONCLUSIONS: Patients want to be treated as unique individuals. They ask for more dialogues with the staff. Awareness of their supposed lifelong fragility implies that health care staff acquires an open and holistic approach. PMID- 15886057 TI - Introducing the Cardibra: a randomised pilot study of a purpose designed support bra for women having cardiac surgery. AB - BACKGROUND: Cardiac nurses instruct women to wear a brassiere (bra) after a sternotomy for cardiac surgery to reduce lateral tension on the wound. However this practice is rarely implemented because regular bras impede nursing care in the immediate postoperative period. The Cardibra was developed to address the inadequacies of the regular bra and provide an alternate method of breast support. AIM: To develop a purpose designed support bra (the Cardibra) and evaluate its effectiveness on sternal wound healing and the reduction of pain for women who had a sternotomy for cardiac surgery. METHODS: The study consisted of two stages. Stage 1 described the development of the Cardibra. Stage 2 was a pilot study of a clinical trial, prospective, randomised two-group design. The treatment group consisted of 10 women using the Cardibra immediately following surgery and the control group of 10 who wore a regular bra 3 days post surgery. Repeated measures at six time points assessed pain levels and wound healing. RESULTS: This study indicated that the Cardibra might have beneficial therapeutic effects on pain levels and wound healing up to day 14 after cardiac surgery. CONCLUSION: This innovative device may offer therapeutic benefits to women following cardiac surgery. PMID- 15886058 TI - The diversity expression of p62 in digestive system cancers. AB - The expression of p62 autoantigen and the frequency of p62 autoantibody have been reported in hepatocellular carcinoma (HCC) and many types of malignant tumors, respectively, but information regarding to the expression of p62 in other cancer tissues and the association of autoantibody to p62 with tumor behaviors is not available. In the current study, the expression of p62 in tissues and the appearance of p62 autoantibody in sera were detected by immunohistochemical staining and ELISA in four clinical types of digestive system cancers including gastric cancer, esophageal cancer, large intestine cancer and HCC, respectively. Interestingly, the immunohistochemistry staining of p62 has been shown in all of digestive canal tissues (stomach, esophagus, large intestine) including tissues with cancers, beside cancers and from non-malignant patients, whereas the frequencies were 62.5% and 0% in tissues with cancer and beside cancer in patients with HCC, respectively. Importantly, we found that the p62 expression and the frequency of autoantibody to p62 were associated to cell differentiation and tumor metastasis, respectively. These results suggest that the expression of p62 in tissues and the appearance of autoantibody to p62 in sera might be related to cell malignant manifestations. Moreover, p62 autoantibody is a significant marker for the prognosis of cancers and the evaluation of clinical treatments. PMID- 15886059 TI - Cervical neoplasia during pregnancy: diagnosis, management and prognosis. AB - Pregnancy represents an exceptional opportunity for the early diagnosis of cervical cancer since visual inspection, cytological examination and bimanual palpation are considered to be part of routine antenatal care. An abnormal cervical smear should generally be managed as in the non-pregnant state. However, colposcopy and biopsies are mainly intended to exclude invasive disease because a conservative approach is preferred in cases of pre-invasive disease. The only absolute indication for conization in pregnancy is to rule out (micro-)invasive disease or make the diagnosis of invasive carcinoma when such a diagnosis will alter the timing or mode of delivery. Overall, earlier stages of cervical cancer are encountered during pregnancy compared with the general population. Although stage of disease and gestational age will largely influence the timing of the interventions, treatment of invasive cervical cancer is similar to the non pregnant state. In strongly desired pregnancies, the use of neo-adjuvant chemotherapy in order to obtain fetal maturity should be considered and discussed with the patient. Although good evidence supports short-term safety, long-term data regarding the in-utero exposure of cytotoxic drugs need to be consolidated. After stratifying for stage, the outcome is similar to the non-pregnant state. PMID- 15886060 TI - The BNP assay does not identify mild left ventricular diastolic dysfunction in asymptomatic diabetic patients. AB - AIMS: We examined the usefulness of BNP for screening for left ventricular (LV) diastolic dysfunction in a sample of type 2 diabetic patients, without structural heart disorder, who have never presented symptoms or signs of heart failure (HF). METHODS AND RESULTS: Seventy-six consecutive patients admitted to the Outpatient Diabetes Clinic were studied. Blood samples were analyzed using the Triage BNP fluorescence immunoassay (Biosite Diagnostics, La Jolla, CA, USA). Echocardiography examinations were performed, with no knowledge of the BNP value. A total of 39 patients out of 76 (51%) were diagnosed with LV diastolic dysfunction and 23 (30%) with LV hypertrophy. Of the patients with LV diastolic dysfunction, impaired relaxation and pseudonormal pattern accounted for 97 and 3% of the cases, respectively. BNP levels among subjects with LV diastolic dysfunction (26+/-22 pg/ml, n=39) were not significantly different from patients with normal LV function (24+/-23 pg/ml, n=37 pg/ml; Mann-Whitney U-test, Z=-0.4, n.s.). CONCLUSIONS: Our data confirm alarmingly high prevalence of LV diastolic dysfunction in asymptomatic individuals with diabetes. Identification of patients with preclinical diabetic cardiomyopathy should be a research and clinical priority. BNP levels cannot be used to detect mild LV diastolic dysfunction in this subset of patients, which requires Doppler echocardiography to be detected. PMID- 15886061 TI - Substrate preferences and glucose uptake in glibenclamide-resistant Leishmania parasites. AB - Several drug-resistant mammalian cell types exhibit increased glycolytic rates, preferential synthesis of ATP through oxidative phosphorylation, and altered glucose transport. Herein we analyzed the influence of parasite growth phase on energy substrate uptake and use in a Leishmania strain [NR(Gr)] selected for resistance against glibenclamide. Glibenclamide is an ABC-transporter blocker which modulates the function of glucose transporters in some mammalian cells. Our results demonstrate for the first time that compared to glibenclamide-sensitive Leishmania, exponential phase glibenclamide-resistant parasites exhibit decreased use of glucose as energy substrate, decreased glucose uptake and decreased glucose transporter expression. However, compared to glibenclamide-sensitive cells, stationary phase resistant parasites display an increased use of amino acids as energy substrate and an increased activity of the enzymes hexokinase, phosphoglucose isomerase, and especially NAD(+)-linked glutamate dehydrogenase. These results suggest that drug resistance in Leishmania involves a metabolic adaptation that promotes a stage dependent modulation of energy substrate uptake and use as a physiological response to the challenge imposed by drug pressure. PMID- 15886062 TI - Identification and phylogenetic comparison of p53 in two distinct mussel species (Mytilus). AB - The extent to which humans and wildlife are exposed to anthropogenic challenges is an important focus of environmental research. Potential use of p53 gene family marker(s) for aquatic environmental effects monitoring is the long-term goal of this research. The p53 gene is a tumor suppressor gene that is fundamental in cell cycle control and apoptosis. It is mutated or differentially expressed in about 50% of all human cancers and p53 family members are differentially expressed in leukemic clams. Here, we report the identification and characterization of the p53 gene in two species of Mytilus, Mytilus edulis and Mytilus trossulus, using RT-PCR with degenerate and specific primers to conserved regions of the gene. The Mytilus p53 proteins are 99.8% identical and closely related to clam (Mya) p53. In particular, the 3' untranslated regions were examined to gain understanding of potential post-transcriptional regulatory pathways of p53 expression. We found nuclear and cytoplasmic polyadenylation elements, adenylate/uridylate-rich elements, and a K-box motif previously identified in other, unrelated genes. We also identified a new motif in the p53 3'UTR which is highly conserved across vertebrate and invertebrate species. Differences between the p53 genes of the two Mytilus species may be part of genetic determinants underlying variation in leukemia prevalence and/or development, but this requires further investigation. In conclusion, the conserved regions in these p53 paralogues may represent potential control points in gene expression. This information provides a critical first step in the evaluation of p53 expression as a potential marker for environmental assessment. PMID- 15886063 TI - Electrocatalytic activity of DNA on electrodes as an indication of hybridisation. AB - Electron transfer between metal electrodes and ferro/ferricyanide is completely suppressed at low ionic concentration. We describe here a new phenomenon related to this reaction: an immobilisation of thiolated single-stranded DNA on gold electrodes retains this activity at low ionic strength up to the level corresponding to the high ionic strength. In contrast, a hybridisation of the complementary DNA with the thiolated single-stranded DNA followed by a binding onto the electrodes, attenuated the electrocatalytic effect. These effects can be used for discrimination between single-stranded DNA and double-stranded DNA and for semi-quantitative measurement of complementary DNA in a sample. PMID- 15886064 TI - Optimizing processing parameters for signal enhancement of oligonucleotide and protein arrays on ARChip Epoxy. AB - Signal enhancement of oligonucleotide and protein arrays on ARChip Epoxy was achieved by optimizing chip processing parameters. The parameters investigated were fabrication, blocking and guide dot concentration, probe concentration and modification, print buffer, humidity during arraying, slide agitation, spot volume and spotter compatibility. The optimum oligonucleotide concentration was 20 microM, while the optimum protein concentration was 0.05 mg/ml. Amino-modified oligonucleotides were best able to be bound to the resin's epoxy groups at pH 8, whereas thiol-modified oligonucleotides displayed an optimum coupling value of pH 7. So as to avoid background (BG) contamination of probes around bright guide dots, the concentration of fluorescent guide dots was set to 1 muM. The most suitable print buffers for oligonucleotide arrays using both piezo- and contact printing systems proved to be 3 x SSC/1.5 M betaine and commercial ArrayLink. When 0.01% monochlortriazinyl-beta-cyclodextrin sodium salt (MCT) was added, the hybridization signal doubled in strength as compared to plain buffer. The optimum print buffer for proteins was 0.1 N phosphate buffer, pH 8/10% glycerine. The optimum humidity for arraying oligonucleotides was 60% and for proteins 40%. Initially agitating slides for 15 min was found just as effective as agitating slides over the total hybridization period (2.5 h), and this resulted in a three times stronger signal. PMID- 15886065 TI - Excluded volume driven counterion condensation inside nanotubes in a concave electrical double layer model. AB - The physical properties of organic nanotubes attract increasing attention due to their potential benefit in technology, biology and medicine. We study the effect of ion size on the electrical properties of cylindrical nanotubes filled with electrolyte solution within a modified Poisson-Boltzmann (PB) approach. For comparison purposes, small hollow nanospheres filled with electrolyte solution are considered. The finite size of the particles in the inner electrolyte solution is described by the excluded volume effect within a lattice statistics approach. We found that an increased ion size reduces the number of counterions near the charged inner surface of the nanotube, leading to an enlarged electrostatic surface potential. The concentration of counterions close to the inner surface saturates for higher surface charge densities and larger ions. In the case of saturation, the closest counterion packing is achieved, all lattice sites near the surface are occupied and an actual counterion condensation is observed. By contrast, the counterion concentration at the axis of the nanotube steadily increases with increasing surface charge density. This growth is more pronounced for smaller nanotube radii and larger ions. At larger nanotube radii for small ion size counterion condensation may also be observed according to the Tsao criterion, i.e. the counterion concentration at the centre is independent of the number of counterions in the system. With decreasing radius the Tsao condensation effect is shifted towards physiologically unrealistic surface charge densities. PMID- 15886066 TI - Influence of temperature, electrical conductivity, power and pH on ascorbic acid degradation kinetics during ohmic heating using stainless steel electrodes. AB - Degradation kinetics of ascorbic acid was determined in pH 5.7 buffer solution using an isothermal batch ohmic heater with stainless steel electrodes. Variables included in this study were temperature (40, 60 and 80 degrees C); power (0, 100,150 and 300 W); and electrical conductivity (varied using 0.25%, 0.5% and 1.0% NaCl). Ascorbic acid concentration was detected by using a HPLC technique. The results indicate that ascorbic acid degradation can be described successfully by a first order model during both conventional and ohmic heating. The Arrhenius relation showed negative values for temperature coefficient (E(T)) during most ohmic treatments, due to a combination of factors that may alter the reaction mechanism. In particular, it appears that at a given power level, higher electric field strengths are conducive to increased incidence of faradaic reactions. Increasing NaCl concentration appears to significantly influence reaction rates via its influence on dissolved oxygen, and through its participation in electrolytic reactions. Contrary to expectations, increasing temperature tended to significantly reduce reaction rate, likely due to decreased dissolved oxygen concentration at high temperature. The results indicate the importance of using inert electrodes in ohmic heating processes. PMID- 15886067 TI - High light-induced changes of 77 K fluorescence emission of pea thylakoid membranes with altered membrane fluidity. AB - The effect of lipid phase order of isolated thylakoid membranes on fluorescent characteristics of both photosystems during illumination with high light intensity at 22 degrees C and 4 degrees C was investigated. For artificial modification of membrane fluidity two membrane perturbing agents were applied cholesterol and benzyl alcohol. 77 K fluorescence emission and excitation spectra of control, cholesterol- and benzyl alcohol-treated thylakoid membranes were analysed in order to determine the high light-induced changes of emission bands attributed to different chlorophyll-protein complexes-F 735, emitted by photosystem I-light-harvesting complex I; and F 685 and F 695, emitted by photosystem II-light-harvesting complex II. Analysis of emission bands showed that high light treatment leads to a decrease of the area of band at 695 nm and a concomitant increase of intensity of the band at 735 nm. The involvement of different pigment pools (chlorophyll a and chlorophyll b) in the energy supply of both photosystems before and after photoinhibitory treatment was estimated on the basis of excitation fluorescence spectra. The dependence of the ratios F 735/F 685 and the band areas at 685 and 695 nm on the illumination time was studied at both temperatures. Data presented indicate that cholesterol incorporation stabilized the intersystem structure in respect to light-induced changes of fluorescence emission of PSI and PSII. It was shown that the effect of fluid properties of thylakoid membranes on the 77 K fluorescence characteristics of main pigment protein complexes of pea thyalkoid membranes depends on the temperature during high light treatment. PMID- 15886068 TI - Overproduction of DNA polymerase eta does not raise the spontaneous mutation rate in diploid human fibroblasts. AB - Telomerase-immortalized lines of diploid xeroderma pigmentosum variant (XP-V) fibroblasts (XP115LO and XP4BE) were complemented for constitutive or regulated expression of wild-type human DNA polymerase eta (hpol eta). The ectopic gene was expressed from a retroviral LTR at a population average of 34- to 59-fold above the endogenous (mutated) mRNA and high levels of hpol eta were detected by immunoblotting. The POLH cDNA was also cloned downstream from an ecdysone regulated promoter and transduced into the same recipient cells. Abundance of the wild-type mRNA increased approximately 10-fold by addition of ponasterone to the culture medium. Complemented cell lines acquired normal resistance to the cytotoxic effects of UVC, even in the presence of 1mM caffeine. They also tolerated higher levels of UVC-induced template lesions during nascent DNA elongation when compared to normal fibroblasts (NHF). UVC-induced mutation frequencies at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus were measured in the XP115LO+XPV cell line overproducing hpol eta constitutively (E. Bassett, N.M. King, M.F. Bryant, S. Hector, L. Pendyala, S.G. Chaney, M. Cordeiro-Stone, The role of DNA polymerase eta in translesion synthesis past platinum-DNA adducts in human fibroblasts, Cancer Res. 64 (2004) 6469-6475). Induced mutation frequencies were significantly reduced, even below those observed in NHF; however, the average mutation frequency in untreated cultures was about three-fold higher than in the isogenic vector-control cell line. In this study, spontaneous HPRT mutation frequencies were measured at regular intervals, as isogenic fibroblasts either lacking or overproducing hpol eta were expanded for 100 population doublings. The mutation rates estimated from these results were not significantly increased in XP115LO cells expressing abnormal levels of hpol eta, relative to the cells lacking this specialized polymerase. These findings suggest that diploid human fibroblasts with normal DNA repair capacities and intact checkpoints are well protected against the potential mutagenic outcome of overproducing hpol eta, while still benefiting from accurate translesion synthesis of UV-induced pyrimidine dimers. PMID- 15886069 TI - Binding of the UvrB dimer to non-damaged and damaged DNA: residues Y92 and Y93 influence the stability of both subunits. AB - UvrB is the ultimate damage-binding protein in bacterial nucleotide excision repair. Previous AFM experiments have indicated that UvrB binds to a damage as a dimer. In this paper we visualize for the first time a UvrB dimer in a gel retardation assay, with the second subunit (B2) more loosely bound than the subunit (B1) that interacts with the damage. A beta-hairpin motif in UvrB plays an important role in damage specific binding. Alanine substitutions of Y92 or Y93 in the beta-hairpin result in proteins that kill E. coli cells as a consequence of incision in non-damaged DNA. Apparently, both residues are needed to prevent binding of UvrB to non-damaged DNA. The lethality of Y93A results from UvrC mediated incisions, whereas that of Y92A is due to incisions by Cho. This difference could be ascribed to a difference in stability of the B2 subunit in the mutant UvrB-DNA complexes. We show that for 3' incision UvrC needs to displace this second UvrB subunit from the complex, whereas Cho seems capable to incise the dimer-complex. Footprint analysis of the contacts of UvrB with damaged DNA revealed that the B2 subunit interacts with the flanking DNA at the 3' side of the lesion. The B2 subunit of mutant Y92A appeared to be more firmly associated with the DNA, indicating that even when B1 is bound to a lesion, the B2 subunit probes the adjacent DNA for presence of damage. We propose this to be a reflection of the process that the UvrB dimer uses to find lesions in the DNA. In addition to preventing binding to non-damaged DNA, the Y92 and Y93 residues appear also important for making specific contacts (of B1) with the damaged site. We show that the concerted action of the two tyrosines lead to a conformational change in the DNA surrounding the lesion, which is required for the 3' incision reaction. PMID- 15886070 TI - Treatment of multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii pneumonia. PMID- 15886071 TI - Postnatal development of carotid body glomus cell O2 sensitivity. AB - In mammals, the main sensors of arterial oxygen level are the carotid chemoreceptors, which exhibit low sensitivity to hypoxia at birth and become more sensitive over the first few days or weeks of life. This postnatal increase in hypoxia sensitivity of the arterial chemoreceptors, termed "resetting", remains poorly understood. In the carotid body, hypoxia is transduced by glomus cells, which are secretory sensory neurons that respond to hypoxia at higher P(O2) levels than non-chemoreceptor cell types. Maturation or resetting of carotid body O2 sensitivity potentially involves numerous aspects of the O2 transduction cascade at the glomus cell level, including glomus cell neurotransmitter secretion, neuromodulator function, neurotransmitter receptor expression, glomus cell depolarization in response to hypoxia, [Ca2+]i responses to hypoxia, K+ and Ca2+ channel O2 sensitivity and K+ channel expression. However, although progress has been made in the understanding of carotid body development, the precise mechanisms underlying postnatal maturation of these numerous aspects of chemotransduction remain obscure. PMID- 15886072 TI - Skin analysis to determine causative agent in dermal exposure to petroleum products. AB - This study evaluates the usefulness of skin analysis to determine the causative agent in cases of dermal exposure. The study consists of an animal experiment and two human cases. The petroleum components detected at high concentrations in skin samples resembled the composition of those in the corresponding petroleum products. However, the petroleum components in blood were detected at low concentrations and were a different composition. Skin is considered to be an advantageous sample to estimate the petroleum product in clinical and forensic cases of dermal exposure. PMID- 15886073 TI - Susceptibility of actin to modification by 4-hydroxy-2-nonenal. AB - 4-Hydroxy-2-nonenal (HNE), a major lipid peroxidation product, reacts with histidine, lysine or cysteine residues of proteins to form hemiacetal Michael adducts and thus interferes with the functions of the proteins. Here we undertook to identify HNE-modified proteins in the target organ of a ferric nitrilotriacetate (Fe-NTA)-induced renal carcinogenesis model with histidine specific HNEJ-2 antibody. Immunoaffinity column separation and sequencing identified one of the major modified proteins as actin. To further explore the characteristics of actin as an HNE acceptor, we produced four novel monoclonal antibodies against HNE-modified keyhole limpet hemocyanin. All these antibodies (HNEJ-1, 3-5) recognized histidine adducts, but were different from HNEJ-2 in recognizing lysine and cysteine adducts to some extent. Actin, albumin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), metallothionein and superoxide dismutase were treated in vitro with HNE and evaluated with these antibodies. The results revealed that actin was most sensitive to HNE modification and metallothionein most resistant. Furthermore, the residue-specificity of GAPDH was in accord with that shown by our recent mass spectrometry data. Immunohistochemistry with the antibodies revealed cytoplasmic staining with or without nuclear staining in the renal proximal tubules after Fe-NTA administration. The results suggest that actin is a major target protein for HNE modification in vivo, and that our monoclonal antibodies are useful for evaluating the HNE adducts produced. PMID- 15886074 TI - Sensitive method for the quantitative determination of bromocriptine in human plasma by liquid chromatography-tandem mass spectrometry. AB - A sensitive LC-MS-MS assay for the quantitative determination of bromocriptine has been developed and validated and is described in this work. The assay involved the extraction of the analyte from 1 ml of human plasma using a solid phase extraction on Oasis MCX cartridges. Chromatography was performed on a Symmetry C18 (2.1 mm x 100 mm, 3.5 microm) column using a mobile phase consisting of 25:75:01 acetonitrile-water-formic acid with a flow rate of 250 microl/min. The linearity was within the concentration range of 2-500 pg/ml. The lower limit of quantification was 2 pg/ml. This method has been demonstrated to be an improvement over existing methods due to its greater sensitivity and specificity. PMID- 15886075 TI - A new anti-obesity drug treatment: first clinical evidence that, antagonising glutamate-gated Ca2+ ion channels with memantine normalises binge-eating disorders. AB - The regulation of appetite relies on complex hypothalamic neurocircuitry of which the arcuate nucleus, and the hormone leptin play important roles. Arcuate nucleus neurones are essential for the regulation of eating behaviour, but they can be intoxicated by elevated serum levels of the amino acid glutamate (GLU). Neurotoxic effects of GLU are mediated by the N-methyl-D-aspartate receptor (NMDA R). But the neurotoxic effects of GLU can be prevented. Concurrent administration of dizocilpine maleate (MK-801), a selective and highly potent non-competitive NMDA-R antagonist, antagonises GLU-gated Ca2+ ion channels and completely prevents the adverse effects of GLU. Also the non-competitive NMDA-R antagonist memantine displays neuroprotective properties. In view of a previously published hypothesis that human obesity results from chronic over-consumption of GLU, we performed a therapeutic trial in five obese, but otherwise healthy women. Memantine treatment markedly decreased appetite within few hours and complete suppressed the binge-eating disorder within 24 h. Body weight decreased markedly within a few days. The findings strongly support the hypothesis that elevated levels of nutritional GLU play an important role in the pathomechanism of human obesity. We suggest to treat human obesity by protecting the hypothalamic signalling cascade of leptin action with low to moderate affinity, non competitive NMDA-R antagonists that selectively block the GLU-gated Ca2+ ion channels. PMID- 15886076 TI - WITHDRAWN: European heights in the early 18th century. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 15886077 TI - Role of the disulphide bridge in folding, stability and function of porcine odorant binding protein: spectroscopic equilibrium studies on C63A/C155A double mutant. AB - Porcine odorant binding protein (pOBP) contains a single disulphide bridge linking residues Cys63 and Cys155. In order to get information on the role played by this crosslink in determining the structural and functional properties of the protein, we substituted these two Cys residues with two Ala residues by site directed mutagenesis and investigated the changes in folding, stability and functional features, as detected by fluorescence and circular dichroism measurements. In particular, we studied both chemical and thermal unfolding/refolding processes under equilibrium conditions, the first induced by guanidinium hydrochloride and the second by raising the temperature from 15 to 90 degrees C. Chemical unfolding curves, as obtained from intrinsic fluorescence and far-UV circular dichroism data, can be fitted by a simple two-state cooperative sigmoidal function; however, their partial overlap (C(1/2)=0.57+/-0.05 from fluorescence and 0.66+/-0.03 from CD) suggests the formation of an intermediate, which lacks tertiary structural features. Thermal unfolding was found to be reversible if the protein was heated up to 65 degrees C, but irreversible above that temperature because of aggregation. The thermodynamic unfolding parameters of this double mutant protein, when compared to those of the wild type protein, clearly point out the important role played by the disulphide bridge on the stability and function of this protein family and probably of many other lipocalins. PMID- 15886078 TI - Deglycosylation of glucoamylase from Aspergillus niger: effects on structure, activity and stability. AB - A comparative structure-function study was performed to establish possible roles of carbohydrates in stabilization of glycoproteins, using glucoamylase (GA) as a model system. In addition to kinetic properties, stability toward elevated temperatures, extremes of pH, high salt concentrations together with circular dichroism, intrinsic/extrinsic fluorescence studies, proteolysis and affinity for interaction with hydrophobic ligands were investigated. Related to all the main properties examined, with one exception, glycosylation provided improvement in functional characteristics of the enzyme, especially in relation to its thermostability. Results are explained in terms of provision of stabilizing intermolecular interactions by the sugar molecules. The improvement in protein rigidity together with reduction of surface hydrophobicity appear to be especially important in relation to prevention of aggregation, an important mechanism of irreversible thermoinactivation, occurring at elevated temperatures. PMID- 15886079 TI - Internal dynamics and protein-matrix coupling in trehalose-coated proteins. AB - We review recent studies on the role played by non-liquid, water-containing matrices on the dynamics and structure of embedded proteins. Two proteins were studied, in water-trehalose matrices: a water-soluble protein (carboxy derivative of horse heart myoglobin) and a membrane protein (reaction centre from Rhodobacter sphaeroides). Several experimental techniques were used: Mossbauer spectroscopy, elastic neutron scattering, FTIR spectroscopy, CO recombination after flash photolysis in carboxy-myoglobin, kinetic optical absorption spectroscopy following pulsed and continuous photoexcitation in Q(B) containing or Q(B) deprived reaction centre from R. sphaeroides. Experimental results, together with the outcome of molecular dynamics simulations, concurred to give a picture of how water-containing matrices control the internal dynamics of the embedded proteins. This occurs, in particular, via the formation of hydrogen bond networks that anchor the protein surface to the surrounding matrix, whose stiffness increases by lowering the sample water content. In the conclusion section, we also briefly speculate on how the protein-matrix interactions observed in our samples may shed light on the protein-solvent coupling also in liquid aqueous solutions. PMID- 15886080 TI - Routine management of gastro-oesophageal reflux disease by gastroenterologists in France: a prospective observational study. AB - AIMS: To obtain data on routine care of gastro-oesophageal reflux disease by French gastroenterologists. PATIENTS: Five thousand and forty-five adults with gastro-oesophageal reflux disease. METHODS: Patients were recruited by a representative sample of 598 French gastroenterologists. A full disease history, with treatments given/investigations undertaken, was obtained from all patients. The patients' expectations with regard to the improvement of the symptoms' control and the corresponding assessments made by the gastroenterologists were recorded on a 4-point scale. RESULTS: The mean time since diagnosis was approximately 4 years. The most commonly presented symptom was heartburn (79% of patients). A total of 3735 patients (74%) underwent endoscopy because of gastro oesophageal reflux disease. Erosive oesophagitis was present in 38% and Barrett's oesophagus in 8%. Medical therapy for gastro-oesophageal reflux disease was prescribed in 92% of patients, of whom 86% received proton pump inhibitors. Lifestyle advice was given to 73% of patients. Overall, 84% of patients and 72% of physicians expected 'highly' or 'moderately' improved control of gastro oesophageal reflux disease. Gastroenterologists underrated the need for improvement of the symptoms' control as expressed by 'de novo' as well as 'previously diagnosed' patients. CONCLUSIONS: This study provides cross-sectional data on the management of gastro-oesophageal reflux disease by gastroenterologists in France. Most patients have high expectations of an improvement in their management, but gastroenterologists generally underrate these expectations. PMID- 15886081 TI - Infliximab in the treatment of Crohn's disease: predictors of response in an Italian multicentric open study. AB - BACKGROUND: Almost 20% of patients with active Crohn's disease are refractory to conventional therapy. Infliximab is a treatment of proven efficacy in this group of patients and it is not clear which variables predict a good response. AIMS.: To evaluate the role of infliximab looking at the predictors of response in a large series of patients with Crohn's disease. PATIENTS AND METHODS: Five hundred and seventy-three patients with luminal refractory Crohn's disease (Crohn's Disease Activity Index (CDAI)>220-400) (312 patients) or with fistulising disease (190 patients) or both of them (71 patients) were treated with a dose of 5 mg/kg in 12 Italian referral centres. The primary endpoints of the study were clinical response and clinical remission for luminal refractory and fistulising disease. We evaluated at univariable and multivariable analysis the following variables: number of infusions, sex, age at diagnosis, smoking habit, site of disease, previous surgery, extraintestinal manifestations and concomitant therapies, and type of fistulas. RESULTS: Patients with luminal refractory disease: 322 patients (84.1%) had a clinical response and 228 (59.5%) reached clinical remission. Patients with fistulising disease: 187 patients (72%) had a reduction of 50% of the number of fistulas and in 107 (41%) a total closure of fistulas was observed. For luminal disease, single infusion (OR 0.49, 95% CI 0.28-0.86) and previous surgery (OR 0.53, 95% CI 0.30-0.93) predicted a worse response for fistulising disease. Other fistulas responded worse than perianal fistulas (OR 0.57, 95% CI 0.303-1.097). CONCLUSION: In Crohn's disease infliximab is effective in luminal refractory and in fistulising disease. A single infusion and previous surgery predicted a worse response in luminal disease whereas perianal fistulas predicted a better response than other type of fistulas. PMID- 15886082 TI - Focal nodular hyperplasia after busulfan treatment. AB - The case of a 48-year-old woman in whom focal nodular hyperplasia of the liver developed after busulfan therapy was administered for essential thrombocytosis is described. Focal nodular hyperplasia is a reactive disorder related to a circulation disorder. The close temporal relation between the haematological disease, busulfan treatment and the appearance of hyperplastic diseases of the liver in our patient supports the possibility that the association of the events might not be casual. PMID- 15886083 TI - Tetrahymena thermophila. PMID- 15886084 TI - Motion distorts perceived position without awareness of motion. PMID- 15886085 TI - The Schizosaccharomyces pombe imprint--nick or ribonucleotide(s)? PMID- 15886087 TI - Chromosome capture: take me to your kinetochore. AB - To form a proper mitotic spindle, all kinetochores must capture at least one microtubule plus end. A new computational modeling study shows that a search-and capture mechanism based on spatially unbiased microtubule dynamic instability is too slow to account for the experimentally observed rate of chromosome capture. PMID- 15886088 TI - Lightness perception: seeing one color through another. AB - A newly described and dramatic visual illusion suggests that the retinal image is decomposed by the brain into overlapping layers, not into contiguous frameworks of illumination. PMID- 15886089 TI - Peripheral glia: Schwann cells in motion. AB - Neuregulin signaling through ErbB receptors is known to play an essential role in Schwann cell proliferation, survival and myelination. Recent studies in zebrafish provide a peek at living Schwann cells migrating along axons in vivo and suggest that ErbB signaling, while not required for cell movement per se, is required to maintain the directed migration of these cells. PMID- 15886090 TI - Sexual selection: the importance of long-term fitness measures. AB - New results from a 20-year study of free-living song sparrows confirm that attractive males contribute more offspring than less attractive males. They also reveal that the offspring of preferred males produce more descendents themselves. Females prefer males with a large song repertoire, which further work shows is a condition-dependent indicator of male quality. PMID- 15886091 TI - Viral immunity: cross-priming with the help of TLR3. AB - Cross-presentation is important for regulating T-cell responses to exogenous antigens and can maintain tolerance (cross-tolerance) or induce immune responses (cross-priming). Recent exciting results on the role of the Toll-like receptor TLR3 in promoting cross-priming of viral antigens provide new insights into the mechanisms that allow Toll-like receptor signaling to bridge innate and adaptive immune responses. PMID- 15886092 TI - Multisensory integration: strategies for synchronization. AB - Recent studies have shown that our ability to match the timing of inputs from objects that can be both seen and heard is highly adaptable and takes into account target depth and the relatively slow speed of sound. PMID- 15886093 TI - Sensorimotor integration: locating locomotion in neural circuits. AB - Neural components of the circuits that transform sensory cues into changes in motor activities are largely unknown. Several recent studies have now functionally mapped the sensorimotor circuits responsible for locomotion behaviors under defined environmental conditions in the nematode Caenorhabditis elegans. PMID- 15886094 TI - Brain asymmetry: switching from left to right. AB - The relationship between structural and functional asymmetries in the brain remains unclear. A recent report describes a zebrafish mutant that provides us with some enticing clues about this relationship. PMID- 15886095 TI - Actin dynamics: growth from dendritic branches. AB - The dendritic nucleation model was devised to explain the cycle of actin dynamics resulting in actin filament network assembly and disassembly in two contexts--at the leading edge of motile cells and in the actin comet tails of intracellular pathogenic bacteria and viruses. Due to the detailed nature of its biochemical predictions, the model has provided an excellent focus for subsequent experimentation. This review summarizes recent work on actin dynamics in the context of the dendritic nucleation model. One outcome of this research is the possibility that additional proteins, as well as the six proteins included in the original model, might increase the efficiency of dendritic nucleation or modify the resulting actin network. In addition, actin dynamics at the leading edge might be influenced by a second actin filament network, independent of dendritic nucleation. PMID- 15886096 TI - Serotonin signaling is a very early step in patterning of the left-right axis in chick and frog embryos. AB - BACKGROUND: Consistent left-right (LR) asymmetry is a fascinating problem in developmental and evolutionary biology. Conservation of early LR patterning steps among vertebrates as well as involvement of nonprotein small-molecule messengers are very poorly understood. Serotonin (5-HT) is a key neurotransmitter with crucial roles in physiology and cognition. We tested the hypothesis that LR patterning required prenervous serotonin signaling and characterized the 5-HT pathway in chick and frog embryos. RESULTS: A pharmacological screen implicated endogenous signaling through receptors R3 and R4 and the activity of monoamine oxidase (MAO) in the establishment of correct sidedness of asymmetric gene expression and of the viscera in Xenopus embryos. HPLC and immunohistochemistry analysis indicates that Xenopus eggs contain a maternal supply of serotonin that is progressively degraded during cleavage stages. Serotonin's dynamic localization in frog embryos requires gap junctional communication and H,K-ATPase function. Microinjection of loss- and gain-of-function constructs into the right ventral blastomere randomizes asymmetry. In chick embryos, R3 and R4 activity is upstream of the asymmetry of Sonic hedgehog expression. MAO is asymmetrically expressed in the node. CONCLUSIONS: Serotonin is present in very early chick and frog embryos. 5-HT pathway function is required for normal asymmetry and is upstream of asymmetric gene expression. The microinjection data reveal asymmetry existing in frog embryos by the 4-cell stage and suggest novel intracellular 5-HT mechanisms. These functional and localization data identify a novel role for the neurotransmitter serotonin and implicate prenervous serotonergic signaling as an obligate aspect of very early left-right patterning conserved to two vertebrate species. PMID- 15886097 TI - Repulsive interactions shape the morphologies and functional arrangement of zebrafish peripheral sensory arbors. AB - BACKGROUND: Trigeminal sensory neurons detect thermal and mechanical stimuli in the skin through their elaborately arborized peripheral axons. We investigated the developmental mechanisms that determine the size and shape of individual trigeminal arbors in zebrafish and analyzed how these interactions affect the functional organization of the peripheral sensory system. RESULTS: Time-lapse imaging indicated that direct repulsion between growing axons restricts arbor territories. Removal of one trigeminal ganglion allowed axons of the contralateral ganglion to cross the midline, and removal of both resulted in the expansion of spinal cord sensory neuron arbors. Generation of embryos with single, isolated sensory neurons resulted in axon arbors that possessed a vast capacity for growth and expanded to encompass the entire head. Embryos in which arbors were allowed to aberrantly cross the midline were unable to respond in a spatially appropriate way to mechanical stimuli. CONCLUSIONS: Direct repulsive interactions between developing trigeminal and spinal cord sensory axon arbors determine sensory neuron organization and control the shapes and sizes of individual arbors. This spatial organization is crucial for sensing the location of objects in the environment. Thus, a combination of undirected growth and mutual repulsion results in the formation of a functionally organized system of peripheral sensory arbors. PMID- 15886098 TI - RIN1 is an ABL tyrosine kinase activator and a regulator of epithelial-cell adhesion and migration. AB - BACKGROUND: ABL tyrosine kinases control actin remodeling in development and in response to environmental stimuli. These changes affect cell adhesion, cell migration, and cell-cell contact. Little is known, however, about upstream mechanisms regulating ABL protein activation. RESULTS: We report that the RAS effector RIN1 is an activator of ABL tyrosine kinases. RIN1 expression in fibroblasts promotes the formation of membrane spikes; similar effects have been reported for ABL overexpression. RIN1 binds to the ABL SH3 and SH2 domains, and these interactions stimulate ABL2 catalytic activity. This leads to increased phosphorylation of CRK and CRKL, inhibiting these cytoskeletal regulators by promoting intramolecular over intermolecular associations. Activated RAS participates in a stable RAS-RIN1-ABL2 complex and stimulates the tyrosine kinase activation function of RIN1. Deletion of the RAS binding domain (RBD) strongly stimulated the ABL2 activation function of RIN1, suggesting that RAS activation results from the relief of RIN1 autoinhibition. The ABL binding domain of RIN1 (RIN1-ABD) increased the activity of ABL2 immune complexes and purified RIN1-ABD stimulated ABL2 kinase activity toward CRK. Mammary epithelial cells (MECs) from Rin1-/- mice showed accelerated cell adhesion and increased motility in comparison to wild-type cells. Knockdown of RIN1 in epithelial-cell lines blocked the induction of CRKL phosphorylation, confirming that RIN1 normally functions as an inhibitor of cell motility. CONCLUSIONS: RIN1 is a directly binding ABL tyrosine kinase activator in cells as well as in a defined-component assay. In response to environmental changes, this novel signal pathway mediates actin remodeling associated with adhesion and migration of epithelial cells. PMID- 15886099 TI - When hymenopteran males reinvented diploidy. AB - In most plants and animals, a consistent relationship exists between the DNA content of a cell and its metabolic activity. The male-haploid sex determination of Hymenoptera and other arthropods may therefore impose a particular selective pressure upon males, which must evolve adaptations to cope with a genomic DNA reduced by half compared with that of females. Here, we show that a nuclear DNA content similar to that of females is restored in muscles of males in all hymenopteran lineages tested except the most basal one (Xyelidae). This doubling of DNA content in males does not occur in other haplodiploid insects, such as thrips (Thysanoptera) and whiteflies (Sternorrhyncha). These results indicate that this adaptation probably occurred early in hymenopteran history, possibly because males acquired strong flying and dispersal abilities. PMID- 15886100 TI - Efficient chromosome capture requires a bias in the 'search-and-capture' process during mitotic-spindle assembly. AB - The mitotic spindle assembles into a bipolar, microtubule-based protein machine during prometaphase. One proposed mechanism for this process is "search-and capture," in which dynamically unstable microtubules (MTs) search space to capture chromosomes. Although existing theoretical estimates suggest that dynamic instability is efficient enough to allow capture within characteristic mitotic timescales, they are limited in scope and do not address the capture times for realistic numbers of chromosomes. Here we used mathematical modeling to explore this issue. We show that without any bias toward the chromosomes, search-and capture is not efficient enough to explain the typical observed duration of prometaphase. We further analyze search-and-capture in the presence of a spatial gradient of a stabilizing factor that biases MT dynamics toward the chromosomes. We show theoretically that such biased search-and-capture is efficient enough to account for chromosome capture. We also show that additional factors must contribute to accelerate the spindle assembly for cells with large nuclear volumes. We discuss the possibility that a RanGTP gradient introduces a spatial bias into microtubule dynamics and thus improves the efficiency of search-and capture as a mechanism for spindle assembly. PMID- 15886101 TI - The heparan sulfate proteoglycans Dally-like and Syndecan have distinct functions in axon guidance and visual-system assembly in Drosophila. AB - Heparan sulfate proteoglycans (HSPGs), a class of glycosaminoglycan-modified proteins, control diverse patterning events via their regulation of growth-factor signaling and morphogen distribution. In C. elegans, zebrafish, and the mouse, heparan sulfate (HS) biosynthesis is required for normal axon guidance, and mutations affecting Syndecan (Sdc), a transmembrane HSPG, disrupt axon guidance in Drosophila embryos. Glypicans, a family of glycosylphosphatidylinositol (GPI) linked HSPGs, are expressed on axons and growth cones in vertebrates, but their role in axon guidance has not been determined. We demonstrate here that the Drosophila glypican Dally-like protein (Dlp) is required for proper axon guidance and visual-system function. Mosaic studies revealed that Dlp is necessary in both the retina and the brain for different aspects of visual-system assembly. Sdc mutants also showed axon guidance and visual-system defects, some that overlap with dlp and others that are unique. dlp+ transgenes were able to rescue some sdc visual-system phenotypes, but sdc+ transgenes were ineffective in rescuing dlp abnormalities. Together, these findings suggest that in some contexts HS chains provide the biologically critical component, whereas in others the structure of the protein core is also essential. PMID- 15886102 TI - Audiovisual integration of speech falters under high attention demands. AB - One of the most commonly cited examples of human multisensory integration occurs during exposure to natural speech, when the vocal and the visual aspects of the signal are integrated in a unitary percept. Audiovisual association of facial gestures and vocal sounds has been demonstrated in nonhuman primates and in prelinguistic children, arguing for a general basis for this capacity. One critical question, however, concerns the role of attention in such multisensory integration. Although both behavioral and neurophysiological studies have converged on a preattentive conceptualization of audiovisual speech integration, this mechanism has rarely been measured under conditions of high attentional load, when the observers' attention resources are depleted. We tested the extent to which audiovisual integration was modulated by the amount of available attentional resources by measuring the observers' susceptibility to the classic McGurk illusion in a dual-task paradigm. The proportion of visually influenced responses was severely, and selectively, reduced if participants were concurrently performing an unrelated visual or auditory task. In contrast with the assumption that crossmodal speech integration is automatic, our results suggest that these multisensory binding processes are subject to attentional demands. PMID- 15886103 TI - fsi zebrafish show concordant reversal of laterality of viscera, neuroanatomy, and a subset of behavioral responses. AB - Asymmetries in CNS neuroanatomy are assumed to underlie the widespread cognitive and behavioral asymmetries in vertebrates. Studies in humans have shown that the laterality of some cognitive asymmetries is independent of the laterality of the viscera; discrete mechanisms may therefore regulate visceral and neural lateralization. However, through analysis of visceral, neuroanatomical, and behavioral asymmetries in the frequent-situs-inversus (fsi) line of zebrafish, we show that the principal left-right body asymmetries are coupled to certain brain asymmetries and lateralized behaviors. fsi fish with asymmetry defects show concordant reversal of heart, gut, and neuroanatomical asymmetries in the diencephalon. Moreover, the neuroanatomical reversals in reversed fsi fish correlate with reversal of some behavioral responses in both fry and adult fsi fish. Surprisingly, two behavioral asymmetries do not reverse, suggesting that at least two separable mechanisms must influence functional lateralization in the CNS. Partial reversal of CNS asymmetries may generate new behavioral phenotypes; supporting this idea, reversed fsi fry differ markedly from their normally lateralized siblings in their behavioral response to a novel visual feature. Revealing a link between visceral and brain asymmetry and lateralized behavior, our studies help to explain the complexity of the relationship between the lateralities of visceral and neural asymmetries. PMID- 15886104 TI - Fungus-growing termites originated in African rain forest. AB - Fungus-growing termites (subfamily Macrotermitinae, Isoptera) cultivate fungal crops (genus Termitomyces, Basidiomycotina) in gardens inside their colonies. Those fungus gardens are continuously provided with plant substrates, whereas older parts that have been well decomposed by the fungus are consumed (cf.). Fungus-growing termites are found throughout the Old World tropics, in rain forests and savannas, but are ecologically dominant in savannas. Here, we reconstruct the ancestral habitat and geographical origin of fungus-growing termites. We used a statistical model of habitat switching repeated over all phylogenetic trees sampled in a Bayesian analysis of molecular data. Our reconstructions provide strong evidence that termite agriculture originated in African rain forest and that the main radiation leading to the extant genera occurred there. Because extant savanna species are found in most genera, this moreover suggests that the savanna has repeatedly been colonized by fungus growing termites. Furthermore, at least four independent "out-of-Africa" migrations into Asia, and at least one independent migration to Madagascar, have occurred. Although fungus growing by termites is ecologically most successful under the variable, unfavorable conditions of the savanna, it seems to have evolved under the more constant and favorable conditions of the rain forest. PMID- 15886105 TI - Recruitment of Mad2 to the kinetochore requires the Rod/Zw10 complex. AB - Compromising the activity of the spindle checkpoint permits mitotic exit in the presence of unattached kinetochores and, consequently, greatly increases the rate of aneuploidy in the daughter cells. The metazoan checkpoint mechanism is more complex than in yeast in that it requires additional proteins and activities besides the classical Mads and Bubs. Among these are Rod, Zw10, and Zwilch, components of a 700 Kdal complex (Rod/Zw10) that is required for recruitment of dynein/dynactin to kinetochores but whose role in the checkpoint is poorly understood. The dynamics of Rod and Mad2, examined in different organisms, show intriguing similarities as well as apparent differences. Here we simultaneously follow GFP-Mad2 and RFP-Rod and find they are in fact closely associated throughout early mitosis. They accumulate simultaneously on kinetochores and are shed together along microtubule fibers after attachment. Their behavior and position within attached kinetochores is distinct from that of BubR1; Mad2 and Rod colocalize to the outermost kinetochore region (the corona), whereas BubR1 is slightly more interior. Moreover, Mad2, but not BubR1, Bub1, Bub3, or Mps1, requires Rod/Zw10 for its accumulation on unattached kinetochores. Rod/Zw10 thus contributes to checkpoint activation by promoting Mad2 recruitment and to checkpoint inactivation by recruiting dynein/dynactin that subsequently removes Mad2 from attached kinetochores. PMID- 15886106 TI - Myosin VIIA defects, which underlie the Usher 1B syndrome in humans, lead to deafness in Drosophila. AB - In vertebrates, auditory and vestibular transduction occurs on apical projections (stereocilia) of specialized cells (hair cells). Mutations in myosin VIIA (myoVIIA), an unconventional myosin, lead to deafness and balance anomalies in humans, mice, and zebrafish; individuals are deaf, and stereocilia are disorganized. The exact mechanism through which myoVIIA mutations result in these inner-ear anomalies is unknown. Proposed inner-ear functions for myoVIIA include anchoring transduction channels to the stereocilia membrane, trafficking stereocilia linking components, and anchoring hair cells by associating with adherens junctions. The Drosophila myoVIIA homolog is crinkled (ck). The Drosophila auditory organ, Johnston's organ (JO), is developmentally and functionally related to the vertebrate inner ear. Both derive from modified epithelial cells specified by atonal and spalt homolog expression, and both transduce acoustic mechanical energy (and references therein). Here, we show that loss of ck/myoVIIA function leads to complete deafness in Drosophila by disrupting the integrity of the scolopidia that transduce auditory signals. We demonstrate that ck/myoVIIA functions to organize the auditory organ, that it is functionally required in neuronal and support cells, that it is not required for TRPV channel localization, and that it is not essential for scolopidial-cell junction integrity. PMID- 15886107 TI - Subcellular localization determines MAP kinase signal output. AB - The Raf-MEK-ERK MAP kinase cascade transmits signals from activated receptors into the cell to regulate proliferation and differentiation. The cascade is controlled by the Ras GTPase, which recruits Raf from the cytosol to the plasma membrane for activation. In turn, MEK, ERK, and scaffold proteins translocate to the plasma membrane for activation. Here, we examine the input-output properties of the Raf-MEK-ERK MAP kinase module in mammalian cells activated in different cellular contexts. We show that the MAP kinase module operates as a molecular switch in vivo but that the input sensitivity of the module is determined by subcellular location. Signal output from the module is sensitive to low-level input only when it is activated at the plasma membrane. This is because the threshold for activation is low at the plasma membrane, whereas the threshold for activation is high in the cytosol. Thus, the circuit configuration of the module at the plasma membrane generates maximal outputs from low-level analog inputs, allowing cells to process and respond appropriately to physiological stimuli. These results reveal the engineering logic behind the recruitment of elements of the module from the cytosol to the membrane for activation. PMID- 15886108 TI - Role of NAADP and cADPR in the induction and maintenance of agonist-evoked Ca2+ spiking in mouse pancreatic acinar cells. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) and cyclic adenosine diphosphate ribose (cADPR) were first demonstrated to mobilize Ca2+ in sea urchin eggs. In the absence of direct measurements of these messengers, pharmacological studies alone have implicated these molecules as intracellular second messengers for specific cell surface receptor agonists. We now report that in mouse pancreatic acinar cells, cholecystokinin, but not acetylcholine, evokes rapid and transient increases in NAADP levels in a concentration-dependent manner. In contrast, both cholecystokinin and acetylcholine-mediated production of cADPR followed a very different time course. The rapid and transient production of NAADP evoked by cholecystokinin precedes the onset of the Ca2+ signal and is consistent with a role for NAADP in the initiation of the Ca2+ response. Continued agonist-evoked Ca2+ spiking is maintained by prolonged elevations of cADPR levels through sensitization of Ca2+ -induced Ca2+ -release channels. This study represents the first direct comparison of NAADP and cADPR measurements, and the profound differences observed in their time courses provide evidence in support of distinct roles of these Ca2+ -mobilizing messengers in shaping specific Ca2+ signals during agonist stimulation. PMID- 15886110 TI - T-cell activation by lipopeptide antigens. AB - The discovery of the CD1 antigen-presenting system reveals that T cells survey the lipid content of target cells via T-cell receptor (TCR) contact with CD1 bound to lipids, glycolipids and small molecules. Recently, CD1 proteins have been found to present mycobacterial lipopeptides that are involved in scavenging iron from infected cells. The mechanism of lipopeptide antigen presentation by CD1 involves the anchoring of antigens in the hydrophobic binding groove, resulting in exposure of the peptide moiety for TCR contact. These findings expand the range of known antigens for T cells and raise the intriguing possibility that CD1, similar to MHC class I and II molecules, enables T cells to discriminate among peptide sequences. PMID- 15886111 TI - Signaling in B cells via Toll-like receptors. AB - Toll-like receptors (TLRs) and their ligands have emerged as important regulators of immunity, relevant to a wide range of effector responses from vaccination to autoimmunity. The most well-studied ligands of TLRs expressed on B cells include the lipopolysaccharides (for TLR4) and CpG-containing DNAs (for TLR9), which induce and/or co-stimulate B cells to undergo proliferation, class switching and differentiation into antibody-secreting cells. Recent developments in this area include advancements into our understanding of the role of these receptor pathways in B cells, and in particular the relevance of TLR9, which has received substantial attention as both a Th1-like inflammatory immunomodulator and a pathogenic co-stimulator of autoreactive B cell responses. PMID- 15886112 TI - Regulation of B lymphocyte activation by complement C3 and the B cell coreceptor complex. AB - Complement is an essential innate immune mechanism that recognizes and eradicates microbes and associated toxins. In addition, complement receptors (CD21 and CD35) on B cells cooperate with the B-cell antigen receptor (BCR) to efficiently recognize and respond to antigens bearing complement C3d(g). Fixation of C3d(g) to antigen confers adjuvant properties and therefore its deposition may need to be carefully regulated to avoid autoreactivity. CD21 and/or CD35 engagement is nonmitogenic, and B-cell activation via BCR-CD21 coligation is enhanced through the recruitment of CD19. Recent efforts have sought a better understanding of the topological and biochemical properties of BCR and coreceptor (CD19-CD21-CD81) signaling, as well as the context for complement activation in the response to foreign and self antigens. PMID- 15886113 TI - Marginal zone B cells in lymphocyte activation and regulation. AB - Marginal zone (MZ) B cells, together with other strategically located innate cells, constitute the first line of defense against blood-borne microorganisms, viruses and toxins in the spleen. Their fast and efficient protective antibody responses are well characterized; however, much less is known of their interactions with other cell types during immune responses. Recent work has demonstrated that MZ B cells can directly activate T cells; and MZ B cells also interact with other antigen presenting cells, transporting and concentrating antigen during the course of T-dependent and T-independent immune responses. PMID- 15886114 TI - ICAM-1 co-stimulates target cells to facilitate antigen presentation. AB - Adhesion molecules are known to mediate cell-cell interactions, particularly those between T cells and antigen-presenting or target cells. Recent studies identified ICAM-1 as a co-stimulatory ligand that binds to lymphocyte function associated antigen-1 (LFA-1), thereby promoting the activation of T cells. As ICAM-1 is expressed on virtually any cell, it becomes a crucial molecule for the activation of CD8(+) T cells in the absence of co-stimulation provided by CD80 and CD86 molecules. In addition, ICAM-1 might function as cell-surface receptor, capable of initiating intracellular signaling. ICAM-1 is associated with other cell molecules, including MHC-I proteins, and our recent data show that productive engagement of ICAM-1 on target cells leads to recruitment of the MHC-I proteins to the contact area and enhances presentation of cognate peptide MHC-I complexes to cytotoxic T cells. PMID- 15886115 TI - Notch: control of lymphocyte differentiation in the periphery. AB - The complexity of the Notch signalling pathway has impeded progress in understanding its precise role in differentiation processes of the mature peripheral immune system. Nevertheless, both B and T lymphocyte responses are undoubtedly influenced by Notch ligation, and defining the mechanisms by which this is achieved remains a challenge. How the innate-adaptive interface and interactions between lymphocytes can be affected, together with the potential of this pathway for therapeutic intervention remain important and stimulating issues. PMID- 15886116 TI - Vav-family proteins in T-cell signalling. AB - The Vav family proteins (Vav1, Vav2, Vav3) are cytoplasmic guanine nucleotide exchange factors (GEFs) for Rho-family GTPases. T-cell antigen receptor (TCR) signalling results in the tyrosine phosphorylation of Vav proteins and hence their activation. Results from mice deficient in one or more Vav proteins has shown that they play critical roles in T-cell development and activation. Vav1 is required for TCR-induced calcium flux, activation of the ERK MAP kinase pathway, activation of the NF-kappaB transcription factor, inside-out activation of the integrin LFA-1, TCR clustering, and polarisation of the T cell. Although many of these processes may require the GEF activity of Vav1, it is possible that Vav1 also has adaptor-like functions. Recent evidence suggests that Vav1 might also function in the nucleus, where it undergoes arginine methylation. An emerging theme is that Vav proteins may have important functions downstream of receptors other than the TCR, such as integrins and chemokine receptors. PMID- 15886117 TI - CD27 and CD70 in T cell and B cell activation. AB - In vitro work has defined the TNF receptor family member CD27 as a T and B cell co-stimulatory molecule. Its activity is governed by the transient availability of its TNF-like ligand CD70 on lymphocytes and dendritic cells. Recent studies, enforcing or abrogating CD27 function by genetic or protein intervention in mouse models have revealed key contributions of the CD27-CD70 system to effector and memory T cell formation, which is probably based on improved cell survival. The stimulatory effects of CD27 on B cell function appear to oppose those of CD70, which also has a signaling role. Targeting CD27-CD70 for therapy is attractive but should take into account the fact that constitutive CD27 stimulation culminates in lethal immunodeficiency. PMID- 15886118 TI - The role of APRIL and BAFF in lymphocyte activation. AB - The TNF family ligands BAFF (also called BLyS) and APRIL regulate lymphocyte survival and activation. BAFF binds to three receptors, BAFF-R, TACI and BCMA, whereas APRIL interacts with TACI, BCMA and proteoglycans. The contribution of BAFF and APRIL to B-cell and plasma-cell survival, CD154 (CD40L)-independent antibody isotype switching, germinal center maintenance, T-dependent and T independent antibody responses, and T cell co-stimulation are relatively well understood. Constitutive BAFF produced by stromal cells determines the size of the peripheral B cell pool, whereas inducible BAFF produced by myeloid and other cells supports local survival of B lymphocytes and can be associated with development of autoimmunity when deregulated. PMID- 15886119 TI - The role of CD22 and other inhibitory co-receptors in B-cell activation. AB - Inhibitory co-receptors downmodulate B-cell receptor (BCR) signalling by setting a signalling threshold that prevents overstimulation of B cells. Activation of these inhibitory co-receptors occurs by phosphorylation on their cytoplasmic inhibitory immunoreceptor tyrosine-based inhibition motifs (ITIMs), followed by recruitment of the tyrosine phosphatase SHP-1 or the lipid phosphatase SHIP, and depends on their association with the BCR. Recent evidence shows that B-cell signal inhibition is regulated by ligand binding of inhibitory receptors. PMID- 15886120 TI - The primary germinal center response in mice. AB - The germinal center (GC) is an important anatomical site for the development of high affinity antibodies during T-cell dependent B cell responses. Although the importance of the GC response to humoral immunity is well known, much remains to be elucidated about GC induction, maintenance and regulation. Recent studies examining the GC response in mice have identified key molecules expressed on follicular dendritic cells that support the differentiation of GC B cells, revealed essential chemokines that direct the organization of light and dark zones, and demonstrated potentially novel roles for TNF family members in the differentiation of GC B cells. PMID- 15886121 TI - Lymphocyte effector mechanisms in innate and adaptive immunity. PMID- 15886122 TI - The dialogue between human natural killer cells and dendritic cells. AB - The interaction of NK cells with dendritic cells (DCs) appears to play an important role in both innate and adaptive immune responses to pathogens. In peripheral inflamed tissues the simultaneous engagement of receptors for danger (e.g. Toll-like receptors), which are expressed by both NK cells and DCs, results in cell activation and the acquisition of functional properties necessary for controlling, and possibly rapidly eliminating, pathogens by innate effector mechanisms. Moreover, NK cells are needed to select the most appropriate DCs that display the functional properties suitable for subsequent T-cell priming. This NK cell-mediated programming of DC maturation is modulated by cytokines released during the early stages of inflammatory responses (i.e. IL-12, IFN-gamma, IL-4). NK cells and DCs continue their interactions in secondary lymphoid organs where both cell types play a role in the control of T-cell priming. PMID- 15886123 TI - Human cytolytic T lymphocytes expressing HLA class-I-specific inhibitory receptors. AB - MHC class-1-specific inhibitory receptors were originally described in NK cells, in which they represent an important fail-safe mechanism that induces NK cell tolerance to normal self cells. These inhibitory NK receptors (iNKRs) were subsequently found expressed on different T cell subsets, primarily CD8(+) cytolytic T lymphocytes (CTLs), in which they can inhibit T cell receptor mediated functions. Some iNKR(+) CTLs are HLA-E-restricted, represent oligo- or monoclonal expansions, and can play a defensive role in viral infections. Although T cell activation, in the presence of certain cytokines, can induce the expression of the CD94-NKG2A heterodimeric receptor, the mechanism leading to the expression of killer immunoglobulin-like receptors (KIRs) is still unknown. The expression of iNKRs in T cells might contribute to the prevention of apoptotic cell death, thus allowing their survival and clonal expansion in vivo. In addition, iNKR(+) T cells might contribute to peripheral self-tolerance. PMID- 15886124 TI - Human tumor-specific T lymphocytes: does function matter more than number? AB - In recent years, several clinical trials have involved the vaccination of cancer patients with tumor-specific antigens that are recognized by T lymphocytes. Anti vaccine T-cell responses in these patients have been monitored on the assumption that their magnitude would correlate with clinical efficacy. Although analysis of these data show that such a correlation is emerging, detailed analyses of the few patients who benefit clinically from the vaccinations suggest that the function of the anti-vaccine T cells might be more important than their number. Recent studies show that in cancer patients numerous tumor-specific T cells appear to be quiescent in the presence of the tumor. Understanding how an efficient vaccine interferes with this coexistence is one of the current challenges of cancer immunotherapy. PMID- 15886125 TI - Understanding the generation and function of memory T cell subsets. AB - Memory T cells can be broadly divided into central memory and effector memory subsets, which are endowed with different capacities to home to lymphoid or non lymphoid tissues, to proliferate in response to antigen or cytokines and to perform effector functions. In the past few years progress has been made in understanding the properties of these memory T cell subsets and, in particular, the signals required for their generation and maintenance. Collectively these data point to a critical role of central memory T cells in conferring long-term immunity. PMID- 15886126 TI - A comparison of gait in spinal muscular atrophy, type II and Duchenne muscular dystrophy. AB - This study investigated and compared the gait of two patients with spinal muscular atrophy, type II (SMA II) and two patients with Duchenne muscular dystrophy (DMD). These diseases cause a progressive and proximal to distal muscular weakness resulting in the loss of ambulation. The DMD cases had comparable muscle weakness with the SMA II cases on manual muscle testing and patients were assessed using kinematics, kinetics, electromyography and video analysis. SMA II and DMD patients employed different gait strategies for forward movement. SMA II patients used pelvic rotation initiated by the upper body to propel the leg forward and produce the necessary step-length whereas the DMD patients tended to use hip flexion and plantar flexion. Management of SMA II patients would include preservation of hip abductor and flexor strength to maintain mobility. PMID- 15886127 TI - Relationship between gait biomechanics and inversion sprains: a prospective study of risk factors. AB - This prospective study determined gait related risk factors for inversion sprains in 223 physical education students. Static lower leg alignment was determined, and 3D-kinematics combined with plantar pressure profiles were collected. After evaluation, the same sports physician registered all sports injuries during the next 6-18 months. During this period, 21 subjects had an inversion sprain, one of whom had a bilateral sprain. Twenty-two ankles, 12 left and 10 right comprised the inversion sprain group and both feet of 36 non-injured subjects acted as controls. Comparison of the two groups revealed that the gait of subjects who are at risk of sustaining an inversion sprain had a laterally situated centre of pressure at initial contact. These subjects also showed a mobile foot type at first metatarsal contact, forefoot flat and heel off. In this type the foot is more pronated over a prolonged period and accompanied by more pressure underneath the medial side of the foot and a delayed maximal knee flexion. Resupination is delayed and roll off does not occur across the hallux, but more laterally, probably because of the diminished support at the first metatarsophalangeal joint. Total foot contact time was also longer in the inversion sprain group compared with controls. The findings of this study suggest that effective prevention and rehabilitation of inversion sprains should include attention to gait patterns and adjustments of foot biomechanics. PMID- 15886128 TI - Push-off reactions in recovery after tripping discriminate young subjects, older non-fallers and older fallers. AB - Tripping is a major cause for falls, especially in the elderly. This study investigated whether falls in the elderly can be attributed to inadequate push off reactions by the support limb in the recovery after a trip. Twelve young (20 34 years) and eleven older (65-72 years) men and women walked over a platform and were tripped several times over an obstacle that suddenly appeared from the floor. Kinematics and ground reactions forces of the support limb during push-off were measured of falls and successful recoveries. Young subjects did not fall. The older subjects were divided into a group of four non-fallers and seven fallers. Older fallers showed insufficient reduction of the angular momentum during push-off and less proper placement of the recovery limb. This was due to a lower rate of change of moment generation in all support limb joints and a lower peak ankle moment. Onset of knee moment generation was slightly delayed in older fallers. Improvement over trials was ascribed to better positioning of the recovery limb, as no clear differences were seen in the joint moments of the support limb. In conclusion, the contribution of the support limb to prevent a fall after tripping is decreased in older adults. Lower limb strength could be an underlying factor and strength training might help to reduce fall risk. PMID- 15886129 TI - Is the method of signal analysis and test selection important for measuring standing balance in subjects with persistent whiplash? AB - Dizziness and or unsteadiness, associated with episodes of loss of balance, are frequent complaints in those suffering from persistent problems following a whiplash injury. Research has been inconclusive with respect to possible aetiology, discriminative tests and analyses used. The aim of this pilot research was to identify the test conditions and the most appropriate method for the analysis of sway that may differentiate subjects with persistent whiplash associated disorders (WAD) from healthy controls. The six conditions of the Clinical Test for Sensory Interaction in Balance was performed in both comfortable and tandem stance in 20 subjects with persistent WAD compared to 20 control subjects. The analyses were carried out using a traditional method of measurement, total sway distance, to results obtained from the use of wavelet analysis. Subjects with WAD were significantly less able to complete the tandem stance tests on a firm surface than controls. In comfortable stance, using wavelet analysis, significant differences between subjects with WAD and the control group were evident in total energy of the trace for all test conditions apart from eyes open on the firm surface. In contrast, the results of the analysis using total sway distance revealed no significant differences between groups across all six conditions. Wavelet analysis may be more appropriate for detecting disturbances in balance in whiplash subjects because the technique allows separation of the noise from the underlying systematic effect of sway. These findings will be used to direct future studies on the aeitiology of balance disturbances in WAD. PMID- 15886130 TI - Changes in the standing posture of stroke patients during rehabilitation. AB - The aim of this study was to analyze the changes in the standing center of pressure (COP) in a hemiplegic adult population at the beginning and the end of in-patient rehabilitation. The trajectory of the COP was recorded on a force platform and was analyzed in terms of the frequency spectra of the center of gravity (COG), and the frequency spectra of the COP minus COG, a derived measure of neuromuscular stiffness. The study population consisted of eight hemiplegic subjects, median age of 53.5 years (range 27-79 years). The median interval between stroke and the first series of measures was 31 days (range 4-127) and the median interval between the two measures was 47.5 days (range 12-92). All the subjects were treated in an inpatient setting in a Neurorehabilitation Center. Our results showed that following rehabilitation, there was a reduction in the amplitude of the medio-lateral frequency spectra of the COP from a median of 5.651 mm (range 3.13-14.05) at the first measurement to 4.408 mm (range 2.40 8.58) at the second measurement. These changes were significant (p<0.05). Significant changes were also observed in the measure of COP minus COG, from 1.324 mm (range 0.92-2.63) on the first measure to 0.917 mm (range 0.46-1.53) on the second measure. In conclusion, the medio-lateral COP trajectory of hemiplegic subjects significantly improved during rehabilitation. We hypothesized that a major component of this improvement was the decrease in neuromuscular stiffness of the adductor and abductor muscles of the hips. PMID- 15886131 TI - Direct measurement of ankle stiffness during quiet standing: implications for control modelling and clinical application. AB - In this study, we describe a device for the direct measurement of intrinsic ankle stiffness in quiet standing. It consists of a motorised footplate mounted on a force platform. By generating random sequences of step-like disturbances (1 degrees amplitude, 150 ms duration) and measuring the corresponding displacements of the center of pressure in the antero-posterior direction, we obtained torque rotation patterns after aligning, averaging, and scaling the postural responses. Such patterns were used for estimating the value of the ankle stiffness, which was normalized as a fraction of the critical value. In order to be confident that the measurements addressed the intrinsic ankle stiffness and were not affected in a significant way by the reflex activation of the muscles in response to the test disturbances, we performed the estimates in different ways: least squares estimates with time windows of different widths and an instantaneous estimate at the time in which the angular acceleration vanishes. The statistical analysis showed that there is no significant difference among the different methods of estimate and the inspection of the electromyographic activity in response to the perturbations showed that at least two of the estimates were certainly outside the possible influence of reflex patterns. The intrinsic ankle stiffness was evaluated to be 64+/-8% of the critical stiffness for test disturbances of the order of 1 degrees. We argue that this figure identifies the lower bound of the range of values which characterise normal sway in quiet standing, whereas the upper bound is given by the estimates performed with much smaller test disturbances [1] which yield a higher value: 91+/-23%. The two estimation paradigms (with very small and very large test disturbances, respectively) are complementary also because they behave in a different way as regards the sensitivity to a bias torque: it is close to zero in the Loram & Lakie's paradigm, whereas it is significant in our paradigm. Thus, as the bias grows, it appears that the range of stiffness values is narrowed and is pushed towards the upper bound. There is a clear potential for the clinical application of these methods, in the sense that the identification of the range of stiffness values used by a patient is a measurable index of motor organisation/reorganisation. PMID- 15886132 TI - Sequencing sit-to-stand and upright posture for mobility limitation assessment: determination of the timing of the task phases from force platform data. AB - The identification of quantitative tools to assess an individual's mobility limitation is a complex and challenging task. Several motor tasks have been designated as potential indicators of mobility limitation. In this study, a multiple motor task obtained by sequencing sit-to-stand and upright posture was used. Algorithms based on data obtained exclusively from a single force platform were developed to detect the timing of the motor task phases (sit-to-stand, preparation to the upright posture and upright posture). To test these algorithms, an experimental protocol inducing predictable changes in the acquired signals was designed. Twenty-two young, able-bodied subjects performed the task in four different conditions: self-selected natural and high speed with feet kept together, and self-selected natural and high speed with feet pelvis-width apart. The proposed algorithms effectively detected the timing of the task phases, the duration of which was sensitive to the four different experimental conditions. As expected, the duration of the sit-to-stand was sensitive to the speed of the task and not to the foot position, while the duration of the preparation to the upright posture was sensitive to foot position but not to speed. In addition to providing a simple and effective description of the execution of the motor task, the correct timing of the studied multiple task could facilitate the accurate determination of variables descriptive of the single isolated phases, allowing for a more thorough description of the motor task and therefore could contribute to the development of effective quantitative functional evaluation tests. PMID- 15886133 TI - Temporal characteristics of foot roll-over during barefoot jogging: reference data for young adults. AB - The purpose of this study was to establish a representative reference dataset for temporal characteristics of foot roll-over during barefoot jogging, based on plantar pressure data collected from 220 healthy young adults. The subjects ran at 3.3 ms-1 over a 16.5 m long running track, having a built-in pressure platform mounted on a force platform. The initial contact, final contact, time to peak pressure and the duration of contact at the lateral and medial heel, metatarsal heads I to V and the hallux were measured. Temporal plantar pressure variables were found to be reliable (93% of ICC coefficients above 0.75) and both gender and asymmetry influences could be neglected. Foot roll-over during jogging started with heel contact followed by a latero-medial contact of the metatarsals and finally the hallux. After heel off, the forefoot started to push off at the lateral metatarsals, followed by a more central push off over the second metatarsal and finally over the hallux. Based on the plantar pressure data, the stance phase during running was divided into four distinct phases: initial contact (8.2%), forefoot contact (11.3%), foot flat (25.3%) and forefoot push off (55.1%). These findings provide a reliable and representative reference dataset for temporal characteristics of foot roll-over during jogging of young adults that may also be relevant in the evaluation of running patterns. PMID- 15886134 TI - Effect of ageing on the ability to adapt to a visual distortion during walking. AB - Degradation of major sensory systems such as proprioception, the vestibular system and vision may be a factor that contributes to the decline in walking stability in older people. In the present study this was examined by introducing a visual distortion by means of prism glasses shifting subject's view 10 degrees to the right while subjects walked towards a target (exposure condition). Shifting the view while walking towards a target will cause subjects to alter their heading in such a way that their walking trajectory describes a curvilinear path. It was expected that older people, when compared to young people, would have greater difficulty adjusting their heading and would show a greater decrease in heading stability, quantified by means of the standard deviation of the lateral position (SDLP). This was indeed the case. When performance in a pre- and post-exposure condition, in which subjects walked without prism glasses, were compared to each other, older people (O group) showed a greater decrease in heading stability than young people (Y group) and middle aged people (M group). Furthermore, it appeared that during the exposure condition adaptation effects were present in the Y and M group, which were absent in the O group. It is discussed that this adaptation is a form of realignment of the proprioceptive and visual system. The absence of realignment in the O group is argued to be caused by degradation of the proprioceptive system, which results in a lowering of the proprioceptive bias of vision. PMID- 15886135 TI - The evolution of clinical gait analysis part III--kinetics and energy assessment. AB - Historically, clinical applications of measurements of force and energy followed electromyography and kinematics in temporal sequence. This sequence is mirrored by the order of topics included in this trilogy on the Evolution of Clinical Gait Analysis, with part I [Sutherland DH. The evolution of clinical gait analysis part I: kinesiological EMG. Gait Posture 2001;14:61-70.] devoted to Kinesiological EMG and part II [Sutherland DH. The evolution of clinical gait analysis part II - kinematics. Gait Posture 2002;16(2):159-179.] to Kinematics. This final review in the series will focus on kinetics as it relates to gait applications. Kinematic measurements give the movements of the body segments, which can be compared with normal controls to identify pathological gait patterns, but they do not deal with the forces controlling the movements. As a major goal of scientifically minded clinicians is to understand the biomechanical forces producing movements, the objective measurement of ground reaction forces is essential. The force plate (platform) is now an indispensable tool in a state of-the-art motion analysis laboratory. Nonetheless, it is not a stand-alone instrument as both kinematic and EMG measurements are needed for maximum clinical implementation and interpretation of force plate measurements. The subject of energy assessment is also given mention, as there is a compelling interest in whether walking has been made easier with intervention. The goals of this manuscript are to provide a historical background, recognize some of the important contributors, and describe the current multiple uses of the force plate in gait analysis. The widespread use of force plates for postural analyses is an important and more recent application of this technology, but this review will be restricted to measurements of gait rather than balance activities. Finally, this manuscript presents my personal perspective and discusses the developments and contributors that have shaped my thoughts and actions, and which I have found to be particularly noteworthy or intriguing. Just as in parts I and II, emphasis has been placed on the early development. All subtopics and important contributors, in this third and certainly most challenging of the review papers, have not been included. Some may find that my perceptions are incomplete. I accept responsibility for all deficiencies, as none were intended. Letters to selected contributors and their responses reveal how each contributor built on the work of others. The level of cooperation and sharing by these early investigators is extraordinary. Had they wished to withhold information about their own work, clinical gait analysis would have been severely delayed. PMID- 15886136 TI - Comments on "The role of the human cerebellum in short- and long-term habituation of postural responses". PMID- 15886138 TI - Therapeutic landscapes: an evolving theme. PMID- 15886139 TI - Healing in places of decline: (re)imagining everyday landscapes in Hamilton, Ontario. AB - Ongoing interest in therapeutic landscapes has contributed noticeably to the development of a "post-medical geography of health" (Kearns, R.A., Professional Geographer 45 (1993) 139). Drawing on a variety of sources, including in-depth interviews and newspaper coverage from Hamilton, Canada, this paper explores the processes by which ordinary places are characterised as healthy or unhealthy, and investigates how health-affirming and health-denying places exist together in everyday life. We argue that it is possible for places to simultaneously hurt and heal, and that the therapeutic effect of place is largely contingent on individuals' physical and social locations. Further, we attempt to illustrate how these meanings are negotiated at a variety of different geographic scales. PMID- 15886140 TI - Medicine, landscapes, symbols: "The country Doctor" by Honore de Balzac. AB - This paper considers examples from research that has interpreted literary works including representations of topographical landscapes, in order to examine the geographical and social perspectives of the author, and of the society in which the author was living and working. We focus especially on the use of representations of therapeutic landscapes in literature to convey the author's perceptions of the aspects of social and physical environments that contribute to human well-being. The novel entitled The country Doctor (Le Medecin de campagne) by Honore de Balzac, first published in 1833, is the particular focus of this paper and we consider this book in the context of the historical period when it was written and other novels of the period which also portray doctors and medicine in varying ways. We present an interpretation of Balzac's novel which focuses on four themes arising from the discussion in the introduction of this paper: the symbolic use of ideas of 'therapeutic landscapes' to represent the 'heroic' role of the doctor; the link between landscapes and ideas of medicine and health, corresponding to the Hippocratic heritage; the representation, through images of salubrious landscapes, of planning and economic development and the role of the leader; the significance of the contrast between the 'therapeutic' rural world and the unhealthy urban settings found in Paris. We conclude that The country Doctor is interesting because of the ways that it reflects the three aspects of landscape (physical, social and symbolic) that Gesler (1998. In: Kearns, R., Gesler W. (Eds.), Putting Health into Place: Landscape, Identity and Well-Being. Syracuse University Press, Syracuse, NY, pp. 17-35 (Chapter 2)) has identified as central 'therapeutic' qualities. Not only does the book reflect a conceptual model of what a therapeutic landscape might be, but it also uses this landscape symbolically as an allegory for spiritual progress and redemption, and for medical and social advance. The novel represents a period of transition from traditional social values and prescientific medical theory towards modernization of society associated with the growing domination of medical science, which continued into the 20th century. PMID- 15886141 TI - Transitory spaces of care: serving homeless people on the street. AB - Within a rapidly expanding body of work exploring the role of hostels and day centres in the accommodation and care of homeless people, very little attention has been paid to the dynamics of the soup run. Soup runs have, however, recently become a focus of concern for the British Government who, echoing 19th century debates regarding the 'inappropriate' distribution of alms, argue that they are undermining attempts to reduce levels of rough sleeping by making it easier for people to survive on the streets. Drawing upon a postal survey, together with a series of interviews and participant observations, this paper develops an in depth account of soup runs in Britain and explores the dynamics of the spaces involved. It argues that far from simply sustaining street homelessness, soup runs provide an important yet very complex series of spaces of care in the contemporary city. By their very nature, however-having a non-interventionist ethos, being transitory, and open to the public eye-the dynamics of these spaces differ in significant ways from those typical of geographically fixed spaces of care. PMID- 15886142 TI - Landscape, care and the relational self: therapeutic encounters in rural England. AB - Over the last decade a number of studies have employed notions of therapeutic landscape to describe the ways in which places become implicated in processes of healing or health enhancement. While this work has usefully highlighted the environmental, social and symbolic dimensions of such places, relatively less consideration has been given to the relational dynamics through which these therapeutic effects emerge. In this paper I seek to address this absence through engagement with two related bodies of work: ecological formulations of place and relational notions of selfhood. These ideas are explored with reference to the experiences of guests at a respite care centre in Dorset, a predominantly rural county in southern England. Alongside its residential services, this centre places a strong emphasis on facilitating guests' engagement with the wider natural environment in which it is set. A number of general analytical and methodological points are developed with regard to future therapeutic landscape research. PMID- 15886143 TI - The political ecology of health: perceptions of environment, economy, health and well-being among 'Namgis First Nation. AB - Informed by Mayer's (Progr. Hum. Geogr 20 (1996) 441) political ecology of disease framework, this paper investigates First Nation's perceptions of the links between environment, economy and health and well-being. A case study of 'Namgis First Nation (Alert Bay, British Columbia, Canada) is used to explore the risks and benefits of salmon aquaculture for British Columbia's First Nations. Analysis of interview data (n = 23) indicates strong links between reduced access to environmental resources, marginal participation in the economy, and declining community health and well being. Results suggest that aquaculture development has further decreased the community's access to environmental resources, thereby restricting those economic, social, and cultural activities that determine good health and well-being for this community. PMID- 15886144 TI - Meeting health need, accessing health care: the role of neighbourhood. AB - Much of what we know about the determinants of access to health care comes from studies undertaken at a large scale, such as between cities, regions/counties/provinces/states and countries. This paper examines local level variations in access to and utilization of health care services across four distinct neighbourhoods in Hamilton, Ontario, Canada. Survey data (n = 1500) were analysed using logistic regression to explore the potential relationships between neighbourhood and health care utilization and unmet health care need. Results show some relationships between neighbourhood of residence and levels of reported utilization as well as unmet need, even when controlling for predisposing, enabling, and need factors (i.e. Age, gender, household composition, income, education, perceived gp visit time) as well as health status. Findings from this empirical study suggest a finer lens is required to examine the mechanisms through which place impacts access to and utilization of care, one that recognizes the roles of compositional, contextual and collective aspects of neighbourhood. PMID- 15886145 TI - The place of motorsport in public health: an Australian perspective. AB - The sport of motor racing provides various public health messages and impacts. Positive messages include road safety campaigns aimed at discouraging drink driving. Negative messages include the sponsorship of racing teams by cigarette companies. Locating motorsport events in significant public places can magnify any health messages or impacts of motorsport in two ways. First, the total local impact of the motor racing events (e.g. through pollution or disruption to healthy modes of transport) is greater in city street circuits than in dedicated racing circuits away from urban areas. Second, the symbolic characteristics of locations with special significance and meaning can enhance the impacts on health by adding legitimacy to the events, thus helping to promote products or activities that either undermine or support public health. This paper explores the health messages and impacts of major motorsport events in Australia, and argues that the distinctive geographies of Australian motorsport work to exacerbate the negative impacts of motorsport on public health. PMID- 15886146 TI - Yielding the bully pulpit: a conversation with Tommy Thompson. Interview by John K Iglehart. AB - In January 2005 Tommy Thompson relinquished the post of secretary of the U.S. Department of Human Services (HHS). Known as being an outspoken defender of his principles throughout his tenure at HHS, Thompson continues that tone in an interview conducted shortly after he left Washington. Topics covered include ways to curb Medicare spending; devote greater attention to preventive health and proper incentives for technology; improve the vaccine infrastructure of the United States and the world; and continue to infuse U.S. foreign policy with what Thompson has termed "medical diplomacy," in an effort to counter the messages of America's enemies. PMID- 15886147 TI - Financing vaccines: in search of solutions that work. PMID- 15886150 TI - Immunizations in the United States: success, structure, and stress. AB - Immunization is a great success of preventive medicine. In the United States, most vaccine-preventable diseases of childhood are at or near record lows while the number of diseases preventable by vaccination has increased. These successes result from a comprehensive system that includes basic research; developing and testing vaccine candidates; a manufacturing base; a regulatory authority; development of immunization policies; implementation of immunization recommendations; and a compensation system for the few people unavoidably injured by vaccines. Despite the successes, the system faces numerous challenges, including vaccine supply, cost, and safety; adult immunization; vaccine research and development; and biopreparedness. PMID- 15886151 TI - The history of vaccines and immunization: familiar patterns, new challenges. AB - Human beings have benefited from vaccines for more than two centuries. Yet the pathway to effective vaccines has been neither neat nor direct. This paper explores the history of vaccines and immunization, beginning with Edward Jenner's creation of the world's first vaccine for smallpox in the 1790s. We then demonstrate that many of the issues salient in Jenner's era-such as the need for secure funding mechanisms, streamlined manufacturing and safety concerns, and deep-seated public fears of inoculating agents-have frequently reappeared and have often dominated vaccine policies. We suggest that historical awareness can help inform viable long-term solutions to contemporary problems with vaccine research, production, and supply. PMID- 15886152 TI - Why are pharmaceutical companies gradually abandoning vaccines? AB - During the past fifty years, the number of pharmaceutical companies making vaccines has decreased dramatically, and those that still make vaccines have reduced resources to make new ones. Pharmaceutical companies are gradually abandoning vaccines because the research, development, testing, and manufacture of vaccines are expensive and because the market to sell vaccines is much smaller than the market for other drug products. Congressional action could assure both a steady supply of existing vaccines and the promise of vaccines for the future. PMID- 15886153 TI - Why certain vaccines have been delayed or not developed at all. AB - Vaccine development is a long process, with the time from early research to licensure steadily increasing. At one time the process took about ten years; now it takes closer to fifteen to twenty years. The process begins with investigators in universities or biotech firms who have an idea. However, to take things further, there must be a vaccine manufacturer, a regulatory authority ready to give permission for the use of the vaccine, and public health authorities that will recommend and foster vaccination. PMID- 15886154 TI - Factors affecting U.S. manufacturers' decisions to produce vaccines. AB - Recent supply interruptions of childhood vaccines have had negative impacts on U.S. public health policies and vaccine delivery. To understand how manufacturers perceive production incentives and disincentives, the Centers for Disease Control and Prevention (CDC) met with the four pharmaceutical firms that sold vaccines through CDC-negotiated contracts during 2002 and 2003. These meetings shed light on the regulatory burden, high costs of the delay between initial investment and sales, and higher costs of new technologies versus older vaccines. All four manufacturers are investing more in research and development because new technologies have advanced their ability to create vaccines not thought possible before. PMID- 15886155 TI - An HIV vaccine: as we build it, will they come? AB - Early researchers accurately predicted that AIDS would have a globally destructive impact. However, other experts erroneously believed that they would be able to develop a vaccine against the virus in a relatively short period. More than twenty years later, scientists continue to work to achieve this goal. This paper addresses the unique obstacles faced by HIV vaccine researchers. It concludes with recommendations for how policymakers and public health officials could collaborate with researchers to overcome these obstacles and contribute to the discovery of an HIV vaccine that would save millions of lives. PMID- 15886157 TI - Vaccine advance-purchase agreements for low-income countries: practical issues. AB - There are strong theoretical arguments for the creation of advance-purchase agreements to increase incentives for the development and production of vaccines for diseases concentrated in low-income countries. A Center for Global Development working group recently concluded that such agreements could be implemented successfully. We consider the practical economic and legal arrangements for such advance-purchase commitments. We identify several practical issues that we believe the public health and policy community should consider further in the design of an advance-purchase commitment. PMID- 15886158 TI - Overcoming economic barriers to the optimal use of vaccines. AB - Vaccines are among the most cost-effective interventions in health care, but economic factors may interfere with their optimal development and delivery in both industrialized and developing countries. For the United States, making the best use of available vaccines will require increasing the financing for vaccines via the public and private systems. For developing countries, innovative and promising approaches include pull mechanisms to establish predictable demand and push mechanisms such as targeted development programs. Partnerships between philanthropy and public resources have made progress in addressing gaps in vaccine financing and development for developing countries, but much remains to be done. PMID- 15886159 TI - Improving vaccine supply and development: who needs what? AB - Payment for vaccines appears, from recent shortages, to have been inadequate. This paper addresses the roles of various stakeholders in influencing current payment and affecting possible increases. It is argued that the recent problems may have stemmed from undervaluation by government payer-negotiators, by private insurers, and ultimately by consumers themselves. On the supply side, the high profits available to other kinds of drug-firm investments may have inhibited allocation of resources to development of new vaccines, and the low profitability and near-monopoly status of current products may have produced insufficient incentives for producers to protect supply against accidents. PMID- 15886160 TI - The problems and promise of vaccine markets in developing countries. AB - As in the market in North America, major barriers to private investment in the development and production of vaccines exist for markets in developing countries. These include the risks of uncertain funding and demand and the difficulties created by historically low pricing. A number of promising and innovative approaches nonetheless are being explored to increase the incentives and reduce the risks of investing in vaccines for developing countries. These innovations are fueled by the growing recognition of powerful stakeholders that vaccines are a critical technology for ensuring global health. PMID- 15886161 TI - Why sole-supplier vaccine markets may be here to stay. AB - Given the structure of costs, demand, and competition, vaccine markets reach long run equilibrium with one or very few suppliers at any point in time. Sole suppliers are less likely to exit and may have lower total social costs. Vaccine markets are dynamically competitive, with new, superior products displacing older, inferior products. Measures to address short-run supply disruptions include inventories, foreign sourcing, and improved technologies. Increasing the relative prices paid for new vaccines to levels that more closely reflect their social value compared to other new drugs and biologics is essential to achieving appropriate incentives for allocation of pharmaceutical R and D. PMID- 15886162 TI - Encouraging the development of new vaccines. AB - Investment in the development of new vaccines is suboptimal. Changing this situation requires a creative blend of "push" and "pull" strategies. One successful policy model is the Orphan Drug Act, whose key features include large research and development (R and D) tax credits as well as Food and Drug Administration (FDA) counseling and priority review. Such supply-side R and D incentive provisions can be combined with demand-side mandates and vouchers to encourage development of new vaccines. Guaranteed-purchase funds and other pull mechanisms are useful supplementary incentives in the cases of vaccines for bioterrorism and neglected diseases of poverty. PMID- 15886163 TI - Addressing the vaccine financing conundrum. AB - An Institute of Medicine (IOM) committee recommended a dramatic change in the national approach to financing immunizations in the United States. Major features were an insurance mandate to cover immunization, federal government subsidy for the purchase of vaccines, and vouchers for uninsured people. The National Vaccine Advisory Committee (NVAC), while agreeing with many of the IOM observations, did not agree with the recommendations for action. NVAC recommended, among other things, expanding and stabilizing funding through the existing Vaccination Assistance Act (Section 317 of the Public Health Service Act), expansion of the Vaccines for Children program, and assurance of adequate reimbursement for administration of vaccines. PMID- 15886165 TI - Vaccine supply: a cross-national perspective. AB - In U.S. vaccine markets, competing producers with high fixed, sunk costs face relatively concentrated demand. This tends to lead to exit of all but one or very few producers per vaccine. Detailed evidence of exits and shortages in the flu vaccine market demonstrates the importance of high fixed costs, demand uncertainty, and dynamic quality competition. A comparison of vaccine suppliers in four industrialized countries compared with the United States shows that smaller foreign markets often have more and different vaccine suppliers. High, country-specific, fixed costs, combined with price and volume uncertainty, plausibly deters these potential suppliers from attempting to enter the U.S. market. PMID- 15886166 TI - Development of priority vaccines for disease-endemic countries: risk and benefit. AB - Decisions regarding vaccine regulation and use made by institutions in industrialized countries can have an unintended impact on vaccines' availability in disease-endemic countries. However, regulatory and programmatic decision making by such countries, taking into consideration local risks and benefits, requires adequate resources, both human and financial. Such differing risk benefit determinations between industrialized and disease-endemic countries will increase product divergence. We propose a single universal standard for risk assessment, based on maximizing net benefit, and an action plan to improve access to priority vaccines through a more robust determination of risk and benefit. PMID- 15886167 TI - Could it happen here? Vaccine risk controversies and the specter of derailment. AB - Controversy over vaccine safety has achieved high visibility over the past decade. At the same time, however, levels of coverage for routinely recommended childhood vaccines in the United States are at their highest ever. We examine this apparent paradox. We consider the ways in which concerns over vaccine safety have emerged and diffused through the popular media, legislative hearings, and Internet-based activism. As a case study, we review the controversy over the alleged connection between autism and the measles-mumps-rubella (MMR) vaccine and consider why it had a dramatic effect on the vaccine's acceptance in Great Britain but virtually none in the United States. PMID- 15886168 TI - It did happen here: fear and loathing on the vaccine trail. AB - Adverse publicity that placed undue emphasis on a possible connection between autism and the measles-mumps-rubella (MMR) vaccine and vaccines containing thimerosal made parents in the United Kingdom reluctant to allow their children to receive the vaccine. The same concerns have played themselves out in the U.S. Food and Drug Administration's decision to recommend removal of thimerosal from other vaccines, even as the individual autism claims have been rejected. That recommendation, based on unsubstantiated safety concerns, reveals a deep-seated institutional overreaction that is more likely to cost lives than to save them. PMID- 15886169 TI - Development of immunization policy and its implementation in the United Kingdom. AB - The U.K. immunization program is managed through a tightly connected process that links the ways policy is developed, submitted to independent expert review and recommendation, adopted into strategy, and implemented into practice. There is one advisory body for immunization. Its recommendations, once accepted by government, are centrally funded, and vaccines are provided at no cost to recipients. Although the present system has worked well, changes in health care management at the peripheral level mean that the immunization program will need to adapt to retain the health gains achieved. Rates of vaccine-preventable disease are at historically low levels, although challenges related to costs and effectiveness remain. PMID- 15886170 TI - Vaccine policies across the pond: looking at the U.K. and U.S. systems. AB - Major differences exist in the immunization programs of the United Kingdom and the United States. If one believes that most health policy decisions in Western industrialized democracies are political, then many of the differences may seem to reflect the variance in the nature of political systems. However, each program has unique components that appear paradoxical, and what works in one society will not necessarily work in another. Those who seek to substitute portions of one vaccine system with those of another must appreciate the context within which each functions. PMID- 15886171 TI - Future directions in vaccines: the payoffs of basic research. AB - Vaccine development has historically relied on approaches such as live attenuated, subunit, and whole-cell vaccine designs to present antigens to the immune system. These strategies are no longer nimble enough to rapidly address public health threats, particularly emerging infectious diseases. New vaccines will require a strong scientific base partnered with the leveraging of emerging and enabling technologies so that candidate vaccines can be developed more rapidly and with the greatest chance of proving effective. This paper focuses on new strategies, technologies, and immunologic research that will provide important opportunities for the development of new and improved vaccines. PMID- 15886172 TI - Coverage of vaccines in private health plans: what does the public prefer? AB - Underinsurance for recommended vaccines in private health plans may affect 15 percent of children and more than 30 percent of adults. We conducted a nationally representative, Web-based study using health plan vignettes to determine adults' willingness to bear marginally higher plan premiums, to assure plan coverage of new vaccines as they are recommended for children and adults. Our results indicate a broad willingness (more than 75 percent of respondents) to pay the higher premiums. Such willingness was associated strongly with perceptions of vaccines' effectiveness and safety. Policymakers, physicians, and public health officials should examine health plan enrollees' preferences as they consider remedies for vaccine underinsurance. PMID- 15886173 TI - Medicaid cost containment and access to prescription drugs. AB - States have been intensifying their efforts to control rising prescription drug costs in their Medicaid programs. This study examines the effects of five Medicaid cost containment policies on enrollees' perceptions of their ability to get prescription drugs. The results show that enrollees in states that have implemented all or almost all of these five policies have greater problems getting prescription drugs than enrollees in other states encounter. In terms of specific policies, prior authorization and mandatory generic substitutions had the largest effects on access to prescription drugs. PMID- 15886174 TI - Making profits and providing care: comparing nonprofit, for-profit, and government hospitals. AB - Three types of entities-nonprofit, for-profit, and government-own hospitals. Yet we know neither whether hospital types specialize in different medical services nor how service profitability affects specialization. In this econometric analysis of American Hospital Association data for every U.S. urban, acute care hospital (1988-2000), more than thirty services were categorized as relatively profitable, unprofitable, or variable. For-profits are most likely to offer relatively profitable medical services; government hospitals are most likely to offer relatively unprofitable services; nonprofits often fall in the middle. For profits are also more responsive to changes in service profitability than the other two types. PMID- 15886175 TI - Effects of the malpractice crisis on access to and incidence of high-risk procedures: evidence from Florida. AB - There is much debate on how recent increases in medical malpractice premiums affect patients' access to care. We examined activity levels of neurosurgeons and obstetricians, as well as the incidence of high-risk surgery and patients' travel times in Florida, where malpractice insurance premiums have soared since 2000. Compared with 1997-2000, we found that during 2000-2003, many neurosurgeons cut back their volume of brain surgeries and that craniotomy patients traveled longer for care without any significant change in the overall incidence of craniotomies. Women undergoing high-risk deliveries did not see increases in travel times. PMID- 15886176 TI - You gotta draw the line somewhere. PMID- 15886177 TI - Luck of the dying. PMID- 15886178 TI - Americans' responses to the 2004 influenza vaccine shortage. AB - Polling taken during the 2004 influenza vaccine shortage found a high level of public concern but no consensus on who should be responsible for solving the problem. A substantial minority questioned whether the vaccine should be reserved for high-risk groups, while a majority thought that care providers should be responsible for allocating it. Many people did not seek vaccination for reasons beyond the shortage, including concerns about safety and efficacy and the perceived low risk from influenza. Although some have suggested that this might have been a successful case of health care rationing, it might not be generalizable to shortages of vaccines for other diseases. PMID- 15886179 TI - Health information, the HIPAA privacy rule, and health care: what do physicians think? AB - This study examines physicians' attitudes toward key Health Insurance Portability and Accountability Act (HIPAA) Privacy Rule requirements and assesses the effects of their implementation. We found that despite physicians' generally negative views toward the Privacy Rule, they rated organizations implementing more rule requirements better at protecting the privacy of patient records than organizations that have not implemented the requirements. The policy implications of the findings are discussed. PMID- 15886180 TI - Measure, learn, and improve: physicians' involvement in quality improvement. AB - Payers, accreditors, and consumers are using quality improvement (QI) methods, but little is known about whether physicians do so. The results from this 2003 national physician survey indicate that most do not. Physicians do not routinely use data for assessing their performance and are reluctant to share those data. They infrequently participate in redesign activities. Physicians in larger and salaried groups are more likely to be engaged in QI. The science of QI has been "institutionalized" but not yet "professionalized."Accelerating physicians' adoption of and participation in QI requires building the infrastructure to support quality and paying attention to professionalism, knowledge, and skills. PMID- 15886181 TI - Leveraging the power of the media to combat HIV/AIDS. AB - Improving HIV/AIDS prevention efforts, especially those focused on young people, is a key challenge in the fight against the pandemic. Since the mid-1990s the Henry J. Kaiser Family Foundation has partnered with U.S.-based media companies on comprehensive HIV/AIDS campaigns. This paper outlines the unique public education model that it is using, and how Kaiser is expanding this work globally by working with some of the largest media companies in the world to undertake new initiatives in Russia and India. PMID- 15886182 TI - A community expands access to health care: the case of access health in Michigan. AB - Access Health, a Michigan-based "three-share plan," is viewed as a successful community-based approach to expanding health benefits in the workplace. It was the stimulus for recently proposed legislation to federally fund similar plans nationally. The program evolved with the support of the W.K. Kellogg Foundation. Its sustained viability is attributable in part to the creative use of a state statute to draw down federal Medicaid disproportionate-share hospital (DSH) funds. Although it faces obstacles common to programs of its type, the program's greatest financial vulnerability rests on the uncertain continued availability of the monies it uses to subsidize the program. PMID- 15886183 TI - Outcomes. PMID- 15886185 TI - Cost of new drugs. PMID- 15886184 TI - Specialty hospitals, ambulatory surgery centers, and general hospitals: charting a wise public policy course. AB - The number of U.S. specialty hospitals and ambulatory surgery centers has been increasing. Advocates of these facilities believe that they will increase competition and improve health services. General hospitals, however, complain that specialty facilities select only the most profitable patients, which reduces general hospitals' ability to pay for care of the uninsured and other unprofitable services. Physician ownership also raises conflict-of-interest concerns. Congress has enacted a moratorium on payments to new specialty hospitals as it ponders the questions that will determine future policy. Can the competitive playing field be leveled, or will future development of these facilities be highly regulated or banned? PMID- 15886187 TI - CPGs: think core concept. PMID- 15886188 TI - Labor union role in patient safety. PMID- 15886189 TI - What about physician assistants? PMID- 15886190 TI - Performance incentives. PMID- 15886191 TI - Hospital referrals. PMID- 15886192 TI - The yeast F(1)F(0)-ATP synthase: analysis of the molecular organization of subunit g and the importance of a conserved GXXXG motif. AB - The F(1)F(0)-ATP synthase enzyme is located in the inner mitochondrial membrane, where it forms dimeric complexes. Dimerization of the ATP synthase involves the physical association of the neighboring membrane-embedded F(0)-sectors. In yeast, the F(0)-sector subunits g and e (Su g and Su e, respectively) play a key role in supporting the formation of ATP synthase dimers. In this study we have focused on Su g to gain a better understanding of the function and the molecular organization of this subunit within the ATP synthase complex. Su g proteins contain a GXXXG motif (G is glycine, and X is any amino acid) in their single transmembrane segment. GXXXG can be a dimerization motif that supports helix helix interactions between neighboring transmembrane segments. We demonstrate here that the GXXXG motif is important for the function and in particular for the stability of Su g within the ATP synthase. Using site-directed mutagenesis and cross-linking approaches, we demonstrate that Su g and Su e interact, and our findings emphasize the importance of the membrane anchor regions of these proteins for their interaction. Su e also contains a conserved GXXXG motif in its membrane anchor. However, data presented here would suggest that an intact GXXXG motif in Su g is not essential for the Su g-Su e interaction. We suggest that the GXXXG motif may not be the sole basis for a Su g-Su e interaction, and possibly these dimerization motifs may enable both Su g and Su e to interact with another mitochondrial protein. PMID- 15886193 TI - Regulation of smooth muscle-specific gene expression by homeodomain proteins, Hoxa10 and Hoxb8. AB - Smooth muscle cells arise from different populations of precursor cells during embryonic development. The mechanisms that specify the smooth muscle cell phenotype in each of these populations of cells are largely unknown. In many tissues and organs, homeodomain transcription factors play a key role in directing cell specification. However, little is known about how these proteins regulate smooth muscle differentiation. Using degenerate reverse transcription PCR coupled to cDNA library screening we identified two homeodomain proteins, Hoxa10 and Hoxb8, which are expressed in adult mouse smooth muscle tissues. All three of the previously described transcripts of the Hoxa10 gene, Hoxa10-1, Hoxa10-2, and Hoxa10-3, were identified. Hoxa10-1 directly activated the smooth muscle-specific telokin promoter but did not activate the SM22alpha, smooth muscle alpha-actin, or smooth muscle myosin heavy chain promoters. Small interfering RNA-mediated knock-down of Hoxa10-1 demonstrated that Hoxa10-1 is required for high levels of telokin expression in smooth muscle cells from uterus and colon. On the other hand, Hoxb8 inhibited the activity of the telokin, SM22alpha, and smooth muscle alpha-actin promoters. Cotransfection of Hoxa10-1 together with Hoxa10-2 or Hoxb8 suggested that Hoxa10-2 and Hoxb8 act as competitive inhibitors of Hoxa10-1. Results from gel mobility shift assays demonstrated that Hoxa10-1, Hoxa10-2, and Hoxb8 bind directly to multiple sites in the telokin promoter. Mutational analysis of telokin promoter reporter genes demonstrated that the three homeodomain protein binding sites located between -80 and -75, +2 and +6, and +14 and +17 were required for maximal promoter activation by Hoxa10-1 and maximal inhibition by Hoxb8. Together these data demonstrate that the genes encoding smooth muscle-restricted proteins are direct transcriptional targets of clustered homeodomain proteins and that different homeodomain proteins have distinct effects on the promoters of these genes. PMID- 15886194 TI - RECQ1 helicase interacts with human mismatch repair factors that regulate genetic recombination. AB - Understanding the molecular and cellular functions of RecQ helicases has attracted considerable interest since several human diseases characterized by premature aging and/or cancer have been genetically linked to mutations in genes of the RecQ family. Although a human disease has not yet been genetically linked to a mutation in RECQ1, the prominent roles of RecQ helicases in the maintenance of genome stability suggest that RECQ1 helicase is likely to be important in vivo. To acquire a better understanding of RECQ1 cellular and molecular functions, we have investigated its protein interactions. Using a co immunoprecipitation approach, we have identified several DNA repair factors that are associated with RECQ1 in vivo. Direct physical interaction of these repair factors with RECQ1 was confirmed with purified recombinant proteins. Importantly, RECQ1 stimulates the incision activity of human exonuclease 1 and the mismatch repair recognition complex MSH2/6 stimulates RECQ1 helicase activity. These protein interactions suggest a role of RECQ1 in a pathway involving mismatch repair factors. Regulation of genetic recombination, a proposed role for RecQ helicases, is supported by the identified RECQ1 protein interactions and is discussed. PMID- 15886195 TI - An alliance between Ras GTPase-activating protein, filamin C, and Ras GTPase activating protein SH3 domain-binding protein regulates myocyte growth. AB - We have previously reported that Ras GTPase-activating protein (RasGAP) is involved in a pathway that regulates total cellular mRNA and protein synthesis in cardiac myocytes. A yeast two-hybrid screen resulted in identification of filamin C (FLN-C) as one of its targets. Knockdown of RasGAP or FLN-C, or severing their interaction, resulted in down-regulation of the RNA polymerase II kinase, cyclin dependent kinase 7 (Cdk7). This appeared to be provoked by the release of cdk7 mRNA from RasGAP SH3 domain-binding protein, G3BP, and its subsequent degradation. In parallel, myocyte growth was also inhibited. On the other hand, overexpression of RasGAP induced a Cdk7- and FLN-C-dependent growth. Thus, we propose that the physical interaction between RasGAP and FLN-C facilitates an interaction between G3BP and cdk7 mRNA. This results in stabilization of cdk7 mRNA, an increase in its protein, which is required for cell growth. PMID- 15886196 TI - The bacterial YbaK protein is a Cys-tRNAPro and Cys-tRNA Cys deacylase. AB - Bacterial prolyl-tRNA synthetases and some smaller paralogs, YbaK and ProX, can hydrolyze misacylated Cys-tRNA Pro or Ala-tRNA Pro. To assess the significance of this quality control editing reaction in vivo, we tested Escherichia coli ybaK for its ability to suppress the E. coli thymidylate synthase thyA:146CCA missense mutant strain, which requires Cys-tRNA(Pro) for growth in the absence of thymine. Missense suppression was observed in a ybaK deletion background, suggesting that YbaK functions as a Cys-tRNA Pro deacylase in vivo. In vitro studies with the full set of 20 E. coli aminoacyl-tRNAs revealed that the Haemophilus influenzae and E. coli YbaK proteins are moderately general aminoacyl-tRNA deacylases that preferentially hydrolyze Cys-tRNA Pro and Cys-tRNA Cys and are also weak deacylases that cleave Gly-tRNA, Ala-tRNA, Ser-tRNA, Pro-tRNA, and Met-tRNA. The ProX protein acted as an aminoacyl-tRNA deacylase that cleaves preferentially Ala tRNA and Gly-tRNA. The potential of H. influenzae YbaK to hydrolyze in vivo correctly charged Cys-tRNA Cys was tested in E. coli strain X2913 (ybaK+). Overexpression of H. influenzae ybaK decreased the in vivo ratio of Cys-tRNA Cys to tRNA Cys from 65 to 35% and reduced the growth rate of strain X2913 by 30% in LB medium. These data suggest that YbaK-mediated hydrolysis of aminoacyl-tRNA can influence cell growth. PMID- 15886197 TI - Parallel regulation of membrane trafficking and dominant-negative effects by misrouted gonadotropin-releasing hormone receptor mutants. AB - Gonadotropin-releasing hormone (GnRH) receptor mutants from patients with hypogonadotropic hypogonadism are frequently misrouted proteins that exert a dominant-negative (DN) effect on human (h) wild-type (WT) receptor, due to oligomerization and retention in the endoplasmic reticulum. Pharmacologic chaperones restore correct folding, rescuing mutants and WT receptor from this oligomer. Rat WT retains the ability to oligomerize (since human and mouse mutants exert a DN effect on rat (r) WT sequence) but, unlike human or mouse, escapes the DN effect of GnRH receptor (Gn-RHR) mutants because rGnRHR mutants route to the plasma membrane with higher efficiency than mouse or human mutants. These distinct behaviors of mouse and rat GnRHRs (distinguished by only four semi or non-conservative amino acid differences) led us to assess the role of each amino acid. The difference in both routing and the DN effect appears mediated primarily by Ser(216) in the rGnRHR. The homologous amino acid in the hGn-RHR is also Ser and is compensated for by the primate-unique insertion of Lys(191) that, alone, dramatically decreases routing of the receptor. These studies establish the relation between the DN effect and altered receptor trafficking and explain why hGnRHR is more susceptible to defective trafficking by disease-related point mutations than rodent counterparts. PMID- 15886198 TI - Activation and inhibition of anaplastic lymphoma kinase receptor tyrosine kinase by monoclonal antibodies and absence of agonist activity of pleiotrophin. AB - Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase that is transiently expressed in specific regions of the central and peripheral nervous systems, suggesting a role in its normal development and function. The nature of the cognate ligands of ALK in vertebrate is still a matter of debate. We produced a panel of monoclonal antibodies (mAbs) directed against the extracellular domain of the human receptor. Two major species of ALK (220 and 140 kDa) were identified in transfected cells, and the use of our mAbs established that the 140-kDa species results from a cleavage of the 220-kDa form. Two mAbs, in the nm range, induced the differentiation of PC12 cells transiently transfected with ALK. In human embryonic kidney 293 cells stably expressing ALK, these two mAbs strongly activated the receptor and subsequently the mitogen-activated protein kinase pathway. We further showed for the first time that activation of ALK also resulted in a specific activation of STAT3. In contrast, other mAbs presented the characteristics of blocking antibodies. Finally, in these cell systems, a mitogenic form of pleiotrophin, a proposed ligand of ALK, failed to activate this receptor. Thus, in the absence of clearly established ligand(s) in vertebrates, the availability of mAbs allowing the activation or the inhibition of the receptor will be essential for a better understanding of the biological roles of ALK. PMID- 15886199 TI - Rearrangements in thyroid hormone receptor charge clusters that stabilize bound 3,5',5-triiodo-L-thyronine and inhibit homodimer formation. AB - In this study, we investigated how thyroid hormone (3,5',5-triiodo-l-thyronine, T3) inhibits binding of thyroid hormone receptor (TR) homodimers, but not TR retinoid X receptor heterodimers, to thyroid hormone response elements. Specifically we asked why a small subset of TRbeta mutations that arise in resistance to thyroid hormone syndrome inhibit both T3 binding and formation of TRbeta homodimers on thyroid hormone response elements. We reasoned that these mutations may affect structural elements involved in the coupling of T3 binding to inhibition of TR DNA binding activity. Analysis of TR x-ray structures revealed that each of these resistance to thyroid hormone syndrome mutations affects a cluster of charged amino acids with potential for ionic bond formation between oppositely charged partners. Two clusters (1 and 2) are adjacent to the dimer surface at the junction of helices 10 and 11. Targeted mutagenesis of residues in Cluster 1 (Arg338, Lys342, Asp351, and Asp355) and Cluster 2 (Arg429, Arg383, and Glu311) confirmed that the clusters are required for stable T3 binding and for optimal TR homodimer formation on DNA but also revealed that different arrangements of charged residues are needed for these effects. We propose that the charge clusters are homodimer-specific extensions of the dimer surface and further that T3 binding promotes specific rearrangements of these surfaces that simultaneously block homodimer formation on DNA and stabilize the bound hormone. Our data yield insight into the way that T3 regulates TR DNA binding activity and also highlight hitherto unsuspected T3-dependent conformational changes in the receptor ligand binding domain. PMID- 15886200 TI - Interaction of the Salmonella-containing vacuole with the endocytic recycling system. AB - Upon entry of the pathogen Salmonella enterica serovar Typhimurium into host cells, the majority of bacteria reside in a membrane-bound compartment called the Salmonella-containing vacuole (SCV). Previous studies have established that the SCV transiently interacts with early endosomes but only acquires a subset of late endosomal/lysosomal proteins. However, the complete set of interactions between the SCV and the endocytic machinery has yet to be characterized. In this study, we have shown that four characterized regulators of endocytic recycling were present on the SCV after invasion. Interaction kinetics were different for each of the regulators; ARF6 and Rab4 associated immediately, but their presence was diminished 60 min post-infection, whereas syntaxin13 and Rab11 association peaked at 60 min. Using a dominant negative approach, we determined that Rab11 regulates the recycling of CD44 from the vacuole but had no effect on major histocompatibility complex (MHC) class I recycling. In contrast, syntaxin13 regulated the recycling of MHC class I but not of CD44. We also determined that maturation of the SCV, measured by the acquisition of lysosomal associated membrane protein-1, slowed when recycling was impaired. These findings suggest that protein movement through the endocytic recycling system is regulated through at least two concurrent pathways and that efficient interaction with these pathways is necessary for maturation of the Salmonella-containing vacuole. We also demonstrate the utility of using Salmonella invasion as a model of endosomal recycling events. PMID- 15886201 TI - BRCA1/BARD1 ubiquitinate phosphorylated RNA polymerase II. AB - The breast- and ovarian-specific tumor suppressor BRCA1, when associated with BARD1, is an ubiquitin ligase. We have shown here that this heterodimer ubiquitinates a hyperphosphorylated form of Rpb1, the largest subunit of RNA polymerase II. Two major phosphorylation sites have been identified in the Rpb1 carboxyl terminal domain, serine 2 (Ser-2) or serine 5 (Ser-5) of the YSPTSPS heptapeptide repeat. Only the Ser-5 hyperphosphorylated form is ubiquitinated by BRCA1/BARD1. Overexpression of BRCA1 in cells stimulated the DNA damage-induced ubiquitination of Rpb1. Similar to the in vitro reaction, the stimulation of Rpb1 ubiquitination by BRCA1 in cells occurred only on those molecules hyperphosphorylated on Ser-5 of the heptapeptide repeat. In vitro, the carboxyl terminus of BRCA1 (amino acids 501-1863) was dispensable for the ubiquitination of hyperphosphorylated Rpb1. In cells, however, efficient Rpb1 ubiquitination required the carboxyl terminus of BRCA1, suggesting that interactions mediated by this region were essential in the complex milieu of the nucleus. These results link the BRCA1-dependent ubiquitination of the polymerase with DNA damage. PMID- 15886202 TI - Raf kinase inhibitory protein regulates Raf-1 but not B-Raf kinase activation. AB - Raf kinase inhibitory protein (RKIP; also known as phosphatidylethanolamine binding protein or PEBP) is a modulator of the Raf/MAPK signaling cascade and a suppressor of metastatic cancer. Here, we show that RKIP inhibits MAPK by regulating Raf-1 activation; specifically, RKIP acts subsequent to Raf-1 membrane recruitment, prevents association of Raf-1 and p21-activated kinase (PAK), and blocks phosphorylation of the Raf-1 kinase domain by PAK and Src family kinases. Mutation of the PAK and Src phosphorylation sites on Raf-1 to aspartate, a phosphate mimic, prevented RKIP association with or inhibition of Raf-1 signaling. Interestingly, although RKIP can interact with B-Raf, RKIP depletion had no effect on activation of B-Raf. Because c-Raf-1 and B-Raf are both required for maximal MAPK stimulation by epidermal growth factor in neuronal and epithelial cell lines, we determined whether RKIP significantly affects MAPK signaling. In fact, RKIP depletion increased not only the amplitude but also the sensitivity of MAPK and DNA synthesis to epidermal growth factor stimulation by up to an order of magnitude. These results indicate that selective modulation of c-Raf-1 but not B-Raf activation by RKIP can limit the dynamic range of the MAPK signaling response to growth factors and may play a critical role in growth and development. PMID- 15886203 TI - Cross-talk between signaling pathways regulates alternative splicing: a novel role for JNK. AB - The regulation of alternative splicing by extracellular signals represents a key event in the control of gene expression. There is increasing evidence showing that many extracellular cues regulate alternative splicing. Nevertheless, the broad picture regarding the role of different signaling pathways and their interaction remains incomplete. Using the fibronectin gene as a model, we show that a laminin-rich basement membrane regulates the alternative splicing of two out of three regions of the transcript (extra domain I and type III connecting segment) in mammary epithelial cells, through a non-stress c-Jun N-terminal kinase (JNK) signaling pathway. We propose that dephosphorylation of the extracellular signal-regulated kinase is involved in this regulatory process. Furthermore, the laminin-rich basement membrane blocks the effect of a mammary mesenchymal cell-conditioned medium, which stimulates the inclusion of extra domain I and type III connecting segment through a phosphatidylinositol3-kinase dependent cascade, indicating that JNK signaling can inhibit the phosphatidylinositol 3-kinase-mediated splicing regulation. These results implicate JNK in the regulation of alternative splicing and provide new evidence on how extracellular stimuli are converted into changes in splicing patterns, strengthening the view that the control of alternative splicing is as complex and relevant as transcriptional control, together accounting for the spatiotemporal requirements of gene expression. PMID- 15886204 TI - Unraveling the mechanism of a potent transcriptional activator. AB - Despite their enormous potential as novel research tools and therapeutic agents, artificial transcription factors (ATFs) that up-regulate transcription robustly in vivo remain elusive. In investigating an ATF that does function exceptionally well in vivo, we uncovered an unexpected relationship between transcription function and a binding interaction between the activation domain and an adjacent region of the DNA binding domain. Disruption of this interaction leads to complete loss of function in vivo, even though the activation domain is still able to bind to its target in the transcriptional machinery. We propose that this interaction parallels those between natural activation domains and their regulatory proteins, concealing the activation domain from solvent and the cellular milieu until it binds to its transcriptional machinery target. Inclusion of this property in the future design of ATFs should enhance their efficacy in vivo. PMID- 15886205 TI - Rapid turnover of c-FLIPshort is determined by its unique C-terminal tail. AB - The caspase-8 inhibitor c-FLIP exists as two splice variants, c-FLIP(L) and c FLIP(S), with distinct roles in death receptor signaling. The mechanisms determining their turnover have not been established. We found that in differentiating K562 erythroleukemia cells both c-FLIP isoforms were inducibly degraded by the proteasome, but c-FLIP(S) was more prone to ubiquitylation and had a considerably shorter half-life. Analysis of the c-FLIP(S)-specific ubiquitylation revealed two lysines, 192 and 195, C-terminal to the death effector domains, as principal ubiquitin acceptors in c-FLIP(S) but not in c FLIP(L). Furthermore the c-FLIP(S)-specific tail of 19 amino acids, adjacent to the two target lysines, was demonstrated to be the key element determining the isoform-specific instability of c-FLIP(S). Molecular modeling in combination with site-directed mutagenesis demonstrated that the C-terminal tail is required for correct positioning and subsequent ubiquitylation of the target lysines. Because the antiapoptotic operation of c-FLIP(S) was not affected by the tail deletion, the antiapoptotic activity and ubiquitin-mediated degradation of c-FLIP(S) are functionally and structurally independent processes. The presence of a small destabilizing sequence in c-FLIP(S) constitutes an important determinant of c FLIP(S)/c-FLIP(L) ratios by allowing differential degradation of c-FLIP isoforms. The conformation-based predisposition of c-FLIP(S) to ubiquitin-mediated degradation introduces a novel concept to the regulation of the death-inducing signaling complex. PMID- 15886206 TI - Spatial segregation of gamma-secretase and substrates in distinct membrane domains. AB - Gamma-secretase facilitates the regulated intramembrane proteolysis of select type I membrane proteins that play diverse physiological roles in multiple cell types and tissue. In this study, we used biochemical approaches to examine the distribution of amyloid precursor protein (APP) and several additional gamma secretase substrates in membrane microdomains. We report that APP C-terminal fragments (CTFs) and gamma-secretase reside in Lubrol WX detergent-insoluble membranes (DIM) of cultured cells and adult mouse brain. APP CTFs that accumulate in cells lacking gamma-secretase activity preferentially associate with DIM. Cholesterol depletion and magnetic immunoisolation studies indicate recruitment of APP CTFs into cholesterol- and sphingolipid-rich lipid rafts, and co-residence of APP CTFs, PS1, and syntaxin 6 in DIM patches derived from the trans-Golgi network. Photoaffinity cross-linking studies provided evidence for the preponderance of active gamma-secretase in lipid rafts of cultured cells and adult brain. Remarkably, unlike the case of APP, CTFs derived from Notch1, Jagged2, deleted in colorectal cancer (DCC), and N-cadherin remain largely detergent-soluble, indicative of their spatial segregation in non-raft domains. In embryonic brain, the majority of PS1 and nicastrin is present in Lubrol WX soluble membranes, wherein the CTFs derived from APP, Notch1, DCC, and N-cadherin also reside. We suggest that gamma-secretase residence in non-raft membranes facilitates proteolysis of diverse substrates during embryonic development but that the translocation of gamma-secretase to lipid rafts in adults ensures processing of certain substrates, including APP CTFs, while limiting processing of other potential substrates. PMID- 15886207 TI - Structure and mechanism of the alkyl hydroperoxidase AhpC, a key element of the Mycobacterium tuberculosis defense system against oxidative stress. AB - The peroxiredoxin AhpC from Mycobacterium tuberculosis (MtAhpC) is the foremost element of a NADH-dependent peroxidase and peroxynitrite reductase system, where it directly reduces peroxides and peroxynitrite and is in turn reduced by AhpD and other proteins. Overexpression of MtAhpC in isoniazid-resistant strains of M. tuberculosis harboring mutations in the catalase/peroxidase katG gene provides antioxidant protection and may substitute for the lost enzyme activities. We report here the crystal structure of oxidized MtAhpC trapped in an intermediate oligomeric state of its catalytic cycle. The overall structure folds into a ring shaped hexamer of dimers instead of the usual pentamer of dimers observed in other reduced peroxiredoxins. Although the general structure of the functional dimer is similar to that of other 2-Cys peroxiredoxins, the alpha-helix containing the peroxidatic cysteine Cys61 undergoes a unique rigid-body movement to allow the formation of the disulfide bridge with the resolving cysteine Cys174. This conformational rearrangement creates a large internal cavity enclosing the active site, which might be exploited for the design of inhibitors that could block the catalytic cycle. Structural and mutagenesis evidence points to a model for the electron transfer pathway in MtAhpC that accounts for the unusual involvement of three cysteine residues in catalysis and suggests a mechanism by which MtAhpC can specifically interact with different redox partners. PMID- 15886208 TI - Smad4 inhibits tumor growth by inducing apoptosis in estrogen receptor-alpha positive breast cancer cells. AB - Estrogen is a mitogen in most estrogen receptor-alpha (ERalpha)-positive breast cancers. We have found that Smad4, a common signal transducer in the transforming growth factor-beta superfamily, acts as an ERalpha transcriptional corepressor. Here, we show that Smad4 induces apoptosis in ERalpha-positive MCF-7 breast cancer cells, but not in ERalpha-negative MDA-MB-231 cells. Smad4 induced expression of short Bim isoforms (by alternative splicing) and Bax and release of cytochrome c in ERalpha-positive cells only, and expression of these apoptotic marker genes was reduced when ERalpha small interfering RNA was introduced. Notably, Smad4 was able to induce apoptosis in MDA-231 cells with acquired ERalpha expression. Furthermore, Smad4 inhibited ERalpha-positive tumor growth by inducing apoptosis in tumor xenografts in nude mice. The sizes of tumors expressing Smad4 were only one-tenth the size of those expressing green fluorescent protein, whereas in ERalpha-negative cells, Smad4 did not reduce the tumor size. Notably, Smad4 also promoted short Bim isoform and Bax expression and release of cytochrome c only in ERalpha-positive MCF-7 tumor xenografts. Bim was sufficient for induction of apoptosis, and the short form was the most potent inducer. Our results demonstrate that Smad4 induces apoptosis by regulating Bim splicing as an initial intrinsic signal in ERalpha-positive cells. Smad4-induced apoptosis in ERalpha-positive breast cancer cells may explain the invasive nature of ERalpha-negative breast tumors, thereby providing a potential target for breast cancer intervention. PMID- 15886209 TI - LMAN1 and MCFD2 form a cargo receptor complex and interact with coagulation factor VIII in the early secretory pathway. AB - Mutations in LMAN1 (ERGIC-53) and MCFD2 are the causes of a human genetic disorder, combined deficiency of coagulation factor V and factor VIII. LMAN1 is a type 1 transmembrane protein with homology to mannose-binding lectins. MCFD2 is a soluble EF-hand-containing protein that is retained in the endoplasmic reticulum through its interaction with LMAN1. We showed that endogenous LMAN1 and MCFD2 are present primarily in complex with each other with a 1:1 stoichiometry, although MCFD2 is not required for oligomerization of LMAN1. Using a cross-linking immunoprecipitation assay, we detected a specific interaction of both LMAN1 and MCFD2 with factor VIII, with the B domain as the most likely site of interaction. We also present evidence that this interaction is independent of the glycosylation state of factor VIII but requires native calcium concentration in the endoplasmic reticulum. The interaction of MCFD2 with factor VIII appeared to be independent of LMAN1-MCFD2 complex formation. These results suggest that LMAN1 and MCFD2 form a cargo receptor complex and that the primary sorting signals residing in the B domain direct the binding of factor VIII to LMAN1-MCFD2 through calcium-dependent protein-protein interactions. MCFD2 may function to specifically recruit factor V and factor VIII to sites of transport vesicle budding within the endoplasmic reticulum lumen. PMID- 15886210 TI - 2-Arachidonoylglycerol stimulates activator protein-1-dependent transcriptional activity and enhances epidermal growth factor-induced cell transformation in JB6 P+ cells. AB - 2-Arachidonoylglycerol (2-AG) is the most abundant endocannabinoid, and it plays a critical role in cannabinoid receptor-mediated cell signaling. Although 2-AG was shown to induce ERK activation via the cannabinoid receptor 1 (CB1), only a nonspecific CB receptor agonist and antagonist was used in those studies. Whether cannabinoid receptor 2 (CB2) is involved in 2-AG-induced ERK activation is still unclear. Moreover, whether 2-AG is involved in mediation of AP-1 activity and cell transformation is also not known. In the present study, we show that 2-AG stimulates AP-1-dependent transcriptional activity and enhances epidermal growth factor-induced cell transformation in mouse epidermal JB6 P+ Cl41 cells. Using JB6 P+ C141 cells, stably transfected with an AP-1 luciferase reporter, we found that 10 microm 2-AG induced up to a 3-fold stimulation of AP-1 transcriptional activity. The AP-1 stimulation appeared to be mediated by ERK but not JNK or p38 kinase. PD98059, a specific inhibitor of MEK1, almost completely blocked 2-AG induced ERK phosphorylation and AP-1 activation. Using CB1/2-/- murine embryonic fibroblasts, we present the first direct evidence that both cannabinoid receptors 1 and 2 (CB1/2) are involved in 2-AG-induced ERK activation. 2-AG could not stimulate ERK phosphorylation or Fyn kinase activity in dominant negative Fyn. In addition, the Fyn inhibitor PP2 blocked 2-AG-induced Fyn kinase activity and ERK phosphorylation and activity. Small interfering RNA Fyn also suppressed 2-AG induced ERK phosphorylation. Interestingly, 2-AG enhanced epidermal growth factor induced AP-1 DNA binding and cell transformation. Taken together, our data provide direct evidence suggesting that 2-AG may have a novel role in cell transformation and carcinogenesis in a signaling pathway involving CB1/2 and activation of Fyn, ERKs, and AP-1. PMID- 15886212 TI - A 16-year-old nephrotic patient with chest pain. PMID- 15886213 TI - Artifactual hypocalcaemia after intravenous administration of gadodiamide (Omniscan). PMID- 15886214 TI - Large-pore haemodialysis membranes: an efficient tool for rapid removal of vancomycin after accidental overdose. PMID- 15886215 TI - The effects of chronic kidney disease and renal replacement therapy on circulating dendritic cells. AB - BACKGROUND: The mechanisms underlying the immunodeficiency of chronic kidney disease (CKD) are incompletely understood. Recently, we described decreased numbers of myeloid (m) and plasmacytoid (p) dendritic cells (DCs), considered the most important antigen-presenting cells, in peripheral blood of patients on chronic intermittent haemodialysis (CIHD). In this study, we analysed whether this reduction resulted from CKD or from renal replacement therapy (RRT). METHODS: Using flowcytometry, we quantified mDCs and pDCs in peripheral blood of patients maintained on CIHD (n = 37), continuous ambulatory peritoneal dialysis (CAPD; n = 29), and patients with CKD not receiving RRT (n = 37). Twenty-nine healthy volunteers served as controls. RESULTS: Patients with CKD (n = 103) had lower pDC and mDC counts compared with volunteers: 4.2 vs 8.3 and 10.0 vs 13.8 x 10(6) cells/l, respectively (P < or = 0.001). Within the CKD group, pDC counts did not differ between patients on CIHD, CAPD and those not receiving RRT (3.6 vs 5.0 vs 4.9 x 10(6) cells/l, respectively). In the latter group, pDC numbers correlated with the glomerular filtration rate (GFR; Spearman's r = 0.49; P<0.01). In contrast, mDC counts of patients on CIHD were lower compared with patients on CAPD (7.5 vs 10.1 x 10(6) cells/l; P = 0.039) and patients not receiving RRT (13.7 x 10(6) cells/l; P<0.001). Among non-dialyzing patients, no correlation existed between GFR and mDC numbers, which were comparable to those of volunteers, even when only non-dialyzing patients with a GFR below 15 ml/min were analysed. CONCLUSIONS: Circulating DC counts are decreased in patients with CKD; for pDCs, this reduction is primarily related to the loss of GFR, whereas the dialysis treatment appears to affect mDC numbers. PMID- 15886216 TI - Diagnosis and successful management of calciphylaxis in a pancreas-kidney transplant patient. PMID- 15886217 TI - Comparison of continuous and intermittent renal replacement therapy for acute renal failure. AB - BACKGROUND: Mortality rates of critically ill patients with acute renal failure (ARF) requiring renal replacement therapy (RRT) are high. Intermittent and continuous RRT are available for these patients on the intensive care units (ICUs). It is unknown which technique is superior with respect to patient outcome. METHODS: We randomized 125 patients to treatment with either continuous venovenous haemodiafiltration (CVVHDF) or intermittent haemodialysis (IHD) from a total of 191 patients with ARF in a tertiary-care university hospital ICU. The primary end-point was ICU and in-hospital mortality, while recovery of renal function and hospital length of stay were secondary end-points. RESULTS: During 30 months, no patient escaped randomization for medical reasons. Sixty-six patients were not randomized for non-medical reasons. Of the 125 randomized patients, 70 were treated with CVVHDF and 55 with IHD. The two groups were comparable at the start of RRT with respect to age (62+/-15 vs 62+/-15 years, CVVHDF vs IHD), gender (66 vs 73% male sex), number of failed organ systems (2.4+/-1.5 vs 2.5+/-1.6), Simplified Acute Physiology Scores (57+/-17 vs 58+/ 23), septicaemia (43 vs 51%), shock (59 vs 58%) or previous surgery (53 vs 45%). Mortality rates in the hospital (47 vs 51%, CVVHDF vs IHD, P = 0.72) or in the ICU (34 vs 38%, P = 0.71) were independent of the technique of RRT applied. Hospital length of stay in the survivors was comparable in patients on CVVHDF [median (range) 20 (6-71) days, n = 36] and in those on IHD [30 (2-89) days, n = 27, P = 0.25]. The duration of RRT required was the same in both groups. CONCLUSION: The present investigation provides no evidence for a survival benefit of continuous vs intermittent RRT in ICU patients with ARF. PMID- 15886218 TI - Renal effects of treatment with diuretics, octreotide or both, in non-azotemic cirrhotic patients with ascites. AB - BACKGROUND: Diuretic-induced hyperreninaemia is associated with renal dysfunction in cirrhotic patients with ascites, and in turn prevents the use of high doses of diuretics. Furthermore, ample evidence suggests that octreotide can inhibit the activation of the renin-aldosterone axis. The present study investigated the renal effects of the addition of octreotide to furosemide and spironolactone in the treatment of non-azotemic cirrhotic patients with ascites. METHODS: We studied 20 patients treated with furosemide and spironolactone. Of them, 10 (Group 1) discontinued diuretic treatment for 7 days. Thereafter, for 5 days each patient received subcutaneous octreotide 300 microg b.i.d., in 10 patients (Group 2) in addition to their usual diuretics. We collected data on the patients while they received diuretics (both groups), after discontinuation of diuretics (Group 1), and after octreotide administration (both groups). RESULTS: We observed a trend to increase creatinine clearance and a significant reduction in plasma active renin and plasma aldosterone after the discontinuation of diuretics. The subsequent introduction of octreotide reduced glomerular filtration rate, although it significantly decreased plasma active renin and plasma aldosterone. In contrast, the addition of octreotide to diuretic treatment significantly increased glomerular filtration rate, urine volume and sodium excretion. The magnitudes of the decreases in plasma-active renin and aldosterone produced by the combination of octreotide and diuretics were similar to those produced by octreotide alone or by the discontinuation of diuretics. CONCLUSIONS: Octreotide alone does not improve renal function in cirrhotic patients with ascites. On the contrary, adding it to diuretic treatment increases glomerular filtration rate and sodium and water excretion, mainly through the suppression of an activated renin-aldosterone axis. PMID- 15886219 TI - Acute renal failure and hypercalcaemia in a man from Guyana: what is the link? PMID- 15886220 TI - Transarterial embolization of ruptured angiomyolipoma associated with tuberous sclerosis. PMID- 15886221 TI - Secondary causes of restless legs syndrome in older people. AB - BACKGROUND: Secondary causes of restless legs syndrome (RLS) have been reported to be more common in those with late-onset RLS. However, 'late-onset' in previous studies was defined as onset after 45 years. OBJECTIVE: To determine the prevalence of secondary causes of RLS and the relationship between aetiological factors and age of symptom onset in an older population. DESIGN: Prospective study conducted over a 5-year period. PARTICIPANTS: 80 consecutive non-related patients diagnosed with RLS. MEASUREMENTS: Patients were assessed according to a standard protocol. Age at symptom onset, severity of symptoms, neurological findings and laboratory tests were examined. RESULTS: Iron deficiency (serum ferritin <50 ng/ml) was present in 22% of patients with onset before 50 years, 39% of those with onset at 50 to 64 years and 58% in those with onset after 64 years (P = 0.009). Clinical neuropathy was also more common in older-onset patients (P = 0.08). Family history was positive in 39%, 23% and 8% of these groups, respectively (P = 0.008). CONCLUSION: Secondary causes of RLS become more common and a positive family history less common with increased age of symptom onset. PMID- 15886222 TI - Nutrient regulation of PKCepsilon is mediated by leucine, not insulin, in skeletal muscle. AB - Nutrients enhance signaling pathways involved in skeletal muscle growth through an increased rate of protein synthesis. These studies have led to an understanding of the potential role of the mammalian target of rapamycin (mTOR) in this process. However, activation of mTOR cannot account for all the stimulatory effects of nutrients. The purpose of these experiments was to examine the effect of nutrients on the cellular distribution and activation state of novel PKC isoforms (PKCepsilon and PKCdelta) in the gastrocnemius of rats by use of modification state-dependent phosphopeptide-specific antibodies. The phosphorylation of PKCepsilon on the catalytic domain autophosphorylation site (Ser(729)) was elevated during feeding and then returned to basal levels when the feeding period ended. Meal feeding augmented the phosphorylation of the downstream effectors of mTOR, namely S6K1 and 4E-BP1. In contrast, the phosphorylation of PKCdelta on either the catalytic domain autophosphorylation site (Ser(643)) or activation loop site (Thr(505)) was unaffected. Similar results were obtained when animals were given leucine either acutely via gavage or chronically by dietary supplementations. The effect of leucine was not mimicked by injecting animals with insulin but could be induced by gavage with norleucine, a structural analog of leucine that does not increase plasma insulin concentration. Thus rises in insulin secondary to meal intake or leucine gavage are probably not responsible for increased phosphorylation of PKCepsilon in response to meal feeding. Elevating the leucine concentration stimulated the phosphorylation of PKCepsilon in gastrocnemius from perfused hindlimb and caused a shift in the distribution of PKCepsilon from the membrane fraction to the cytosolic fraction. The results indicate that leucine leads to an activation (autophosphorylation) and subcellular redistribution of PKCepsilon, but not PKCdelta, in gastrocnemius both in vivo and in vitro. Furthermore, activation of the mTOR signaling pathway above basal conditions does not appear to be necessary to induce phosphorylation or translocation of PKCepsilon, suggesting that multiple signaling pathways become activated with leucine. PMID- 15886223 TI - Oleylethanolamide impairs glucose tolerance and inhibits insulin-stimulated glucose uptake in rat adipocytes through p38 and JNK MAPK pathways. AB - Oleylethanolamide (OEA) is a lipid mediator that inhibits food intake and body weight gain and also exhibits hypolipemiant actions. OEA exerts its anorectic effects peripherally through the stimulation of C-fibers. OEA is synthesized in the intestine in response to feeding, increasing its levels in portal blood after the meal. Moreover, OEA is produced by adipose tissue, and a lipolytic effect has been found. In this work, we have examined the effect of OEA on glucose metabolism in rats in vivo and in isolated adipocytes. In vivo studies showed that acute administration (30 min and 6 h) of OEA produced glucose intolerance without decreasing insulin levels. Ex vivo, we found that 10 min of preincubation with OEA inhibited 30% insulin-stimulated glucose uptake in isolated adipocytes. Maximal effect was achieved at 1 microM OEA. The related compounds palmitylethanolamide and oleic acid had no effect, suggesting a specific mechanism. Insulin-stimulated GLUT4 translocation was not affected, but OEA promoted Ser/Thr phosphorylation of GLUT4, which may impair transport activity. This phosphorylation may be partly mediated by p38 and JNK kinases, since specific inhibitors (SB-203580 and SP-600125) partly reverted the inhibitory effect of OEA on insulin-stimulated glucose uptake. These results suggest that the lipid mediator OEA inhibits insulin action in the adipocyte, impairing glucose uptake via p38 and JNK kinases, and these effects may at least in part explain the glucose intolerance produced in rats in vivo. These effects of OEA may contribute to the anorectic effects induced by this mediator, and they might be also relevant for insulin resistance in adipose tissue. PMID- 15886224 TI - Long-term caloric restriction increases UCP3 content but decreases proton leak and reactive oxygen species production in rat skeletal muscle mitochondria. AB - Calorie restriction (CR) without malnutrition increases life span and delays the onset of a variety of diseases in a wide range of animal species. However, the mechanisms responsible for the retardation of aging with CR are poorly understood. We proposed that CR may act, in part, by inducing a hypometabolic state characterized by decreased reactive oxygen species (ROS) production and mitochondrial proton leak. Here, we examine the effects of long-term CR on whole animal energetics as well as muscle mitochondrial energetics, ROS production, and ROS damage. CR was initiated in male FBNF1 rats at 6 mo of age and continued for 12 or 18 mo. Mean whole body VO2 was 34.6 (P < 0.01) and 35.6% (P < 0.001) lower in CR rats than in controls after 12 and 18 mo of CR, respectively. Body mass adjusted VO2 was 11.1 and 29.5% lower (both P < 0.05) in CR rats than in controls after 12 and 18 mo of CR. Muscle mitochondrial leak-dependent (State 4) respiration was decreased after 12 mo compared with controls; however, after 18 mo of CR, there were slight but not statistically significant differences. Proton leak kinetics were affected by 12 mo of CR such that leak-dependent respiration was lower in CR mitochondria only at protonmotive force values exceeding 170 mV. Mitochondrial H2O2 production and oxidative damage were decreased by CR at both time points and increased with age. Muscle UCP3 protein content increased with long-term CR, consistent with a role in protection from ROS but inconsistent with the observed decrease or no change in proton leak. PMID- 15886225 TI - Leptin-mediated activation of human platelets: involvement of a leptin receptor and phosphodiesterase 3A-containing cellular signaling complex. AB - An elevated circulating level of the adipocyte-derived satiety hormone leptin is an independent risk factor for cardiovascular disease. Because thrombus formation is a major cause of acute coronary events and leptin was shown previously to facilitate ADP-induced platelet aggregation, we chose to define the signaling events involved in leptin-mediated platelet activation. Using pharmacological, biochemical, and cell biological approaches, we show that leptin-induced platelet activation required activation of a signaling cascade that included the long form of the leptin receptor, three kinases [Janus kinase 2 (JAK2), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (PKB/Akt)], the insulin receptor substrate-1 (IRS-1), and the major human platelet cAMP phosphodiesterase phosphodiesterase 3A (PDE3A). Moreover, we identify a role for an intraplatelet LEPR/JAK2/IRS-1/PI3K/PKB/PDE3A molecular complex that allows for the selective leptin-mediated activation of platelets. Our data demonstrate that leptin promotes platelet activation, provides a mechanistic basis for the prothrombotic effect of this hormone, and identifies a potentially novel therapeutic avenue to limit obesity-associated cardiovascular disease. PMID- 15886226 TI - Initially more rapid small intestinal glucose delivery increases plasma insulin, GIP, and GLP-1 but does not improve overall glycemia in healthy subjects. AB - The rate of gastric emptying of glucose-containing liquids is a major determinant of postprandial glycemia. The latter is also dependent on stimulation of insulin secretion by glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1). Although overall emptying of glucose approximates 1-3 kcal/min, the "early phase" of gastric emptying is usually more rapid. We have evaluated the hypothesis that increased stimulation of incretin hormones and insulin by a more rapid initial rate of small intestinal glucose delivery would reduce the overall glycemic response to a standardized enteral glucose load. Twelve healthy subjects were studied on two separate days in which they received an intraduodenal (id) glucose infusion for 120 min. On one day, the infusion rate was variable, being more rapid (6 kcal/min) between t = 0 and 10 min and slower (0.55 kcal/min) between t = 10 and 120 min, whereas on the other day the rate was constant (1 kcal/min) from t = 0-120 min, i.e., on both days 120 kcal were given. Between t = 0 and 75 min, plasma insulin, GIP, and GLP-1 were higher with the variable infusion. Despite the increase in insulin and incretin hormones, blood glucose levels were also higher. Between t = 75 and 180 min, blood glucose and plasma insulin were lower with the variable infusion. There was no difference in the area under the curve 0-180 min for blood glucose. We conclude that stimulation of incretin hormone and insulin release by a more rapid initial rate of id glucose delivery does not lead to an overall reduction in glycemia in healthy subjects. PMID- 15886227 TI - Inhibition of adipose tissue lipolysis increases intramuscular lipid and glycogen use in vivo in humans. AB - This study investigates the consequences of inhibition of adipose tissue lipolysis on skeletal muscle substrate use. Ten subjects were studied at rest and during exercise and subsequent recovery under normal, fasting conditions (control trial, CON) and following administration of a nicotinic acid analog (low plasma free fatty acid trial, LFA). Continuous [U-13C]palmitate and [6,6-2H2]glucose infusions were applied to quantify plasma free fatty acid (FFA) and glucose oxidation rates and to estimate intramuscular triacylglycerol (IMTG) and glycogen use. Muscle biopsies were collected to measure 1) fiber type-specific IMTG content; 2) allosteric regulators of hormone-sensitive lipase (HSL), glycogen phosphorylase, and pyruvate dehydrogenase; and 3) the phosphorylation status of HSL at Ser563 and Ser565. Administration of a nicotinic acid analog (acipimox) substantially reduced plasma FFA rate of appearance and subsequent plasma FFA concentrations (P < 0.0001). At rest, this substantially reduced plasma FFA oxidation rates, which was compensated by an increase in the estimated IMTG use (P < 0.05). During exercise, the progressive increase in FFA rate of appearance, uptake, and oxidation was prevented in the LFA trial and matched by greater IMTG and glycogen use. Differential phosphorylation of HSL or relief of its allosteric inhibition by long-chain fatty acyl-CoA could not explain the increase in muscle TG use, but there was evidence to support the contention that regulation may reside at the level of the glucose-fatty acid cycle. This study confirms the hypothesis that plasma FFA availability regulates both intramuscular lipid and glycogen use in vivo in humans. PMID- 15886228 TI - Anabolic growth hormone action improves submaximal measures of physical performance in patients with HIV-associated wasting. AB - Growth hormone (GH) treatment reverses the muscle loss allegedly responsible for diminished aerobic capacity and increased fatigue in patients with HIV-associated wasting. This study examined whether submaximal measures of physical performance can be used as objective measures of the functional impact of GH treatment induced anabolism. We randomized 27 HIV-positive men [mean (SD) age, 43.9 (7.2) yr; body mass, 71.9 (10.4) kg; BMI, 23.1 (2.8) kg/m2] with unintentional weight loss despite antiretroviral therapy to receive GH (6 mg) or placebo in a double blinded, placebo-controlled, cross-over trial with a 3-mo washout. Lean body mass (LBM), maximum oxygen uptake (Vo2 peak), ventilatory threshold (VeT), 6-min walk test (6MWT) distance and work, profile of mood states (POMS) fatigue and vigor scores, and Nottingham health profile (NHP) energy and physical mobility scores were measured. LBM significantly increased after 3 mo of GH treatment vs. placebo (means +/- SE, 3.7 +/- 0.6 vs. 0.3 +/- 0.4 kg; P < 0.001). VeT significantly improved (17.6 +/- 3.7 vs. -5.9 +/- 2.5%; P < 0.001), but Vo2 peak did not change significantly. 6MWT distance improved (24.9 +/- 9.7 vs. 19.9 +/- 11.6 m; P > 0.05) and 6MWT work increased significantly more after 3 mo of GH treatment (33.3 +/- 8.8 vs. 16.5 +/- 7.5 kJ; P < 0.05). POMS scores of fatigue and vigor and the NHP score of energy improved, yet the changes were not statistically significant. GH treatment improved VeT linearly to the increase in LBM (r =0.43, P = 0.037) and 6MWT work (r = 0.51, P = 0.008), and the increase in 6MWT work correlated with increase in LBM (r = 0.45, P = 0.024). Improvement in 6MWT work above the median (27.3 kJ) showed a decrease in fatigue (r = -0.62, P = 0.024). We concluded that GH treatment-induced LBM gains in HIV-associated wasting were functionally relevant, as determined by effort-independent submaximal measures of cardiopulmonary exercise testing. PMID- 15886229 TI - Glycogen debranching enzyme association with beta-subunit regulates AMP-activated protein kinase activity. AB - AMP-activated protein kinase (AMPK) regulates both glycogen and lipid metabolism functioning as an intracellular energy sensor. In this study, we identified a 160 kDa protein in mouse skeletal muscle lysate by using a glutathione-S-transferase (GST)-AMPK fusion protein pull-down assay. Mass spectrometry and a Mascot search revealed this protein to be a glycogen debranching enzyme (GDE). The association between AMPK and GDE was observed not only in the overexpression system but also endogenously. Next, we showed the beta1-subunit of AMPK to be responsible for the association with GDE. Furthermore, experiments using deletion mutants of the beta1-subunit of AMPK revealed amino acids 68-123 of the beta1-subunit to be sufficient for GDE binding. W100G and K128Q, both beta1-subunit mutants, are reportedly incapable of binding to glycogen, but both bound GDE, indicating that the association between AMPK and GDE does not involve glycogen. Rather, the AMPK GDE association is likely to be direct. Overexpression of amino acids 68-123 of the beta1-subunit inhibited the association between endogenous AMPK and GDE. Although GDE activity was unaffected, basal phosphorylation and kinase activity of AMPK, as well as phosphorylation of acetyl-CoA carboxylase, were significantly increased. Thus it is likely that the AMPK-GDE association is a novel mechanism regulating AMPK activity and the resultant fatty acid oxidation and glucose uptake. PMID- 15886230 TI - Autoimmune regulator-1 messenger ribonucleic acid analysis in a novel intronic mutation and two additional novel AIRE gene mutations in a cohort of autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy patients. AB - CONTEXT: Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare autosomal recessive disease associated with mutations in the AIRE gene. OBJECTIVE: Our objective was to investigate clinical and mutational characteristics of 12 Slovenian patients from 10 families with APECED. METHODS: Direct sequencing, restriction fragment length polymorphism, and amplification refractory mutation system analyses were used to identify AIRE gene mutations. Autoimmune regulator (AIRE)-1 mRNA analysis was used to confirm pathogenicity of the intronic mutation. RESULTS: The prevalence of APECED in Slovenian population was estimated to be 1 in 43,000, which is significantly higher compared with the neighboring populations. Three novel mutations were identified among six different mutations detected in the AIRE gene. The first novel mutation was an intronic mutation (653-7_-5delCTC) affecting proper splicing by using a nearby new acceptor splice site as demonstrated by AIRE-1 mRNA analyses. The second and third novel mutations were frame-shift mutations located in exon 5 (540delG) and exon 9 (1064-1068dupCCCGG), both leading to premature truncation of the AIRE protein. The Finnish R257X mutation was the most frequent AIRE gene mutation in Slovenian patients with APECED (16 of 24 alleles). CONCLUSIONS: Three novel AIRE gene mutations were identified. For the first time, a novel intronic mutation was investigated on the mRNA level in APECED. This could be particularly important for APECED patients where no or only heterozygous mutation on the genomic DNA level is detected. PMID- 15886231 TI - Short nights attenuate light-induced circadian phase advances in humans. AB - CONTEXT: In humans, sleep duration often determines the night (dark) length experienced, because we close our eyes when we sleep and are exposed to artificial or natural light when we are awake. Although it is recognized that there is an increasing trend in modern society toward shorter sleep time, it is not known how short nights (long photoperiods) affect the human circadian system. OBJECTIVE: In this study we investigated for the first time the effects of night length on circadian phase shifts to light in humans. DESIGN AND SETTING: Eight young healthy subjects experienced 2 wk of 6-h sleep episodes in the dark (short nights) and 2 wk of long 9-h sleep episodes (long nights) in counterbalanced order. After each series of nights, they were exposed to four 30-min pulses of morning bright light (approximately 5000 lux) that advanced by 1 h/d for 3 consecutive days while night (dark) length was maintained at 6 or 9 h. Circadian phase was determined from the circadian rhythm of melatonin in dim light before and after the 3-d bright light treatments. RESULTS: The phase advance in the melatonin rhythm during the short nights was less than half of that observed during the long nights (P < 0.05). CONCLUSIONS: This result shows for the first time that people who curtail their sleep may unwittingly reduce their circadian responsiveness to morning light. This finding also demonstrates that sleep length can alter human circadian function and has important implications for enhancing the treatment of circadian rhythm sleep disorders. PMID- 15886232 TI - Serum 3,3',5'-triiodothyronine (rT3) and 3,5,3'-triiodothyronine/rT3 are prognostic markers in critically ill patients and are associated with postmortem tissue deiodinase activities. AB - INTRODUCTION AND METHODS: Critical illness is associated with reduced TSH and thyroid hormone secretion, and with changes in peripheral thyroid hormone metabolism, resulting in low serum T3 and high rT3. In 451 critically ill patients who received intensive care for more than 5 d, serum thyroid parameters were determined on d 1, 5, 15, and last day (LD). All patients had been randomized for intensive or conventional insulin treatment. Seventy-one patients died, and postmortem liver and skeletal muscle biopsies were obtained from 50 of them for analysis of deiodinase (D1-3) activities. RESULTS: Insulin treatment did not affect thyroid parameters. On d 1, rT3 was higher and T3/rT3 was lower in nonsurvivors as compared with survivors (P = 0.001). Odds ratio for survival of the highest vs. the lowest quartile was 0.3 for rT3 and 2.9 for T3/rT3. TSH, T4, and T3 were lower in nonsurvivors from d 5 until LD (P < 0.001). TSH, T4, T3, and T3/rT3 increased over time in survivors, but decreased or remained unaltered in nonsurvivors. Liver D1 activity was positively correlated with LD serum T3/rT3 (R = 0.83, P < 0.001) and negatively correlated with rT3 (R = -0.69, P < 0.001). Both liver and skeletal muscle D3 activity were positively correlated with LD serum rT3 (R = 0.32, P = 0.02 and R = 0.31, P = 0.03). CONCLUSION: In critically ill patients who required more than 5 d of intensive care, rT3 and T3/rT3 were already prognostic for survival on d 1. On d 5, T4, T3, but also TSH levels are higher in patients who will survive. Serum rT3 and T3/rT3 were correlated with postmortem tissue deiodinase activities. PMID- 15886233 TI - Pituitary tumor transforming gene binding factor: a novel transforming gene in thyroid tumorigenesis. AB - CONTEXT: There are currently no clear markers for the detection of differentiated thyroid cancer and its recurrence. Pituitary tumor transforming gene (PTTG) is a protooncogene implicated in the pathogenesis of multiple tumor types, which stimulates fibroblast growth factor-2 secretion via PTTG binding factor (PBF). OBJECTIVE: The aim of this study was to ascertain whether PBF expression is associated with thyroid cancer outcome. DESIGN: PBF expression was measured at the mRNA and protein level. Tissue was collected during surgery, with normal samples being taken from the contralateral lobe. In vitro studies ascertained the ability of PBF to transform cells and form tumors in nude mice and its subcellular localization. SETTING: The study was conducted at a primary care/referral center. PATIENTS: Thyroid tumors were collected from a series of 27 patients undergoing surgical excision of papillary and follicular thyroid tumors. INTERVENTION: No intervention was conducted. MAIN OUTCOME MEASURE: The expression of PBF in thyroid cancers compared with normal thyroid, hypothesized before the investigation to be raised in tumors, was the main outcome measure. RESULTS: PBF mRNA expression was higher in differentiated thyroid carcinomas than in normal thyroid (P < 0.001; n = 27) and was independently associated with tumor recurrence (P = 0.002; R(2) = 0.49). PTTG was able to up-regulate PBF mRNA expression in vitro (P < 0.001; n = 12), and stable overexpression of PBF in NIH3T3 cells resulted in significant colony formation (P < 0.001; n = 12). In vivo, stable sc overexpression of PBF induced tumor formation in athymic nude mice. CONCLUSIONS: PBF is an additional prognostic indicator in differentiated thyroid cancer that is transforming in vitro and tumorigenic in vivo. PMID- 15886234 TI - EP1 receptor-mediated migration of the first trimester human extravillous trophoblast: the role of intracellular calcium and calpain. AB - CONTEXT: The root cause of preeclampsia in the human lies in the placenta, where a subpopulation of cytotrophoblast cells called extravillous trophoblasts (EVT), known to be involved in the invasion of the uterine endometrium and utero placental arteries, become less invasive, resulting in poor perfusion of maternal blood into placenta. OBJECTIVES: Because EVT migrate into the prostaglandin (PG) E2-rich decidua, we tested the roles of PGE2 and PGE2-mediated signaling in EVT migration, using our well-characterized EVT line HTR-8/Svneo as well as first trimester villus explants in culture. DESIGN: mRNA expression of different PGE2 receptors (EPs) in HTR-8/Svneo cells was studied using RT-PCR. To characterize the functional significance of EP receptors in EVT, different EP receptor agonists and antagonists were used in our migration assay systems and in the measurements of intracellular concentration of Ca2+ ([Ca2+]i) and calpain activity. RESULTS: Exogenous PGE2 stimulated EVT migration both in vitro and in the villus explant cultures. Although EVT expressed mRNA for all EP receptors (EP 1-4), a functional predominance of EP1 and EP4 was demonstrated in migration assays using specific EP agonists and antagonists. EP1-receptor-mediated signaling events such as activation of phospholipase C and elevation of cytosolic free [Ca2+]i were confirmed by the following findings: 1) exogenous PGE2 or an EP1 agonist, but not an EP4 agonist, increased [Ca2+]i, which could be blocked with an EP1 antagonist as well as BAPTA and thapsigargin; 2) phospholipase C inhibitor U73122, BAPTA, and thapsigargin inhibited PGE2-mediated migratory response of EVT; and 3) PGE2-mediated EVT migration was shown to be dependent on a class of Ca2+-dependent proteases called calpains, known to be involved in cell detachment from substratum during migratory responses. The presence of PGE2 stimulated calpain activity, whereas two calpain inhibitors, calpastatin and N-Ac Leu-Leu-methioninal (ALLM), blocked EVT migration. CONCLUSION: PGE2 stimulates EVT migration by signaling through EP1 receptors, increasing [Ca2+]i, and activating calpain. PMID- 15886235 TI - Vitamin D receptor gene polymorphisms are associated with the risk of fractures in postmenopausal women, independently of bone mineral density. AB - CONTEXT: Osteoporosis is a systemic disease with a strong genetic component. Vitamin D receptor (VDR) gene polymorphisms explain only a small part of the genetic influence on the level of bone mineral density (BMD), whereas their effect on fracture remains uncertain. OBJECTIVE: The objective of this study was to investigate the relationships between VDR genotypes and fracture risk. DESIGN: A prospective population-based cohort was studied. SUBJECTS: A total of 589 postmenopausal women (mean age, 62 yr) were followed prospectively during a median (interquartile) of 11 (1.1) yr. MAIN OUTCOME MEASURE: The study measured incidents of vertebral and nonvertebral fractures. RESULTS: VDR allele B was significantly and dose dependently overrepresented in women who fractured, including 34 and 86 women with first incident vertebral and nonvertebral fragility fractures, respectively. This corresponded to an odds ratio of 1.5 (95% confidence interval, 0.95-2.40) for heterozygous carriers (bB, n = 286) and 2.10 (95% confidence interval, 1.16-3.79) for homozygous carriers (BB, n = 90) of the B allele, compared with women with the bb genotype (n = 213). VDR genotype groups did not differ for demographics, physical activity, grip strength, personal and maternal history of fracture, and calcium intake. The association was independent of BMD of the spine, hip, and radius, and of the BMD loss at the radius. The relationship between VDR polymorphisms and fracture risk was not altered after adjustment for baseline circulating levels of bone turnover markers, estradiol, dehydroepiandrosterone sulfate, SHBG, IGF-I, intact PTH, and 25 hydroxyvitamin D. CONCLUSION: VDR genotypes are associated with the risk of fracture in postmenopausal women independently of BMD, rate of postmenopausal forearm BMD loss, bone turnover, and endogenous hormones. The mechanisms by which VDR genotypes influence bone strength remain to be determined. PMID- 15886236 TI - Cortisol response in relation to the severity of stress and illness. AB - BACKGROUND: The aim of the study was to compare the adrenal response, the course of the ACTH/cortisol ratio, as well as the variance and the diagnostic performance of different cutoffs after 1 and 250 microg ACTH stimulation in different stress situations. METHODS: We investigated three groups with increasing stress levels: ambulatory controls (group A; n = 20), hospitalized medical patients (group B; n = 25), and patients undergoing coronary artery bypass grafting (group C; n = 29). All subjects underwent four consecutive ACTH stimulation tests and were randomized to either a 1- or 250-microg dose. RESULTS: Stimulated cortisol levels in group A were similar to basal cortisol levels under maximal stress (C3; P = 0.8). Peak cortisol concentrations were higher after 250 microg compared with 1 microg ACTH in group B (P = 0.006) and under maximal stress after extubation (group C3; P = 0.027), whereas there were no differences in group A. The ACTH/cortisol ratio was lower in surgical patients after extubation compared with unstressed conditions (P < or = 0.03) The within-subject variance was similar in ambulatory controls and medical patients and after both ACTH doses (all 17-36% of total variance). Cutoff dependent, the diagnosis of relative adrenal insufficiency would have been made in 0-58.3%, respectively. CONCLUSION: In moderate and major stress situations, cortisol concentrations in patients without hypothalamic-pituitary-adrenal disease were higher after stimulation with 250 microg compared with 1 mug ACTH. Data from our study give insight into the physiological adaptations of the hypothalamic-pituitary-adrenal axis to stress. PMID- 15886237 TI - Occupational exposure to ionizing radiation is associated with autoimmune thyroid disease. AB - CONTEXT: The thyroid gland is a potential target organ for radiation-related damage. OBJECTIVE: The aim of the analysis was to investigate the association between occupational exposure to ionizing radiation and autoimmune thyroid disease (AITD). DESIGN: Our design was the cross-sectional Study of Health in Pomerania. SETTING: The setting was the general community. SUBJECTS: Analyses were performed in a population-based sample of 4299 subjects. Among them, 160 persons reported a history of occupational exposure to ionizing radiation. MAIN OUTCOME MEASURE: AITD was defined as the combined presence of hypoechogenicity in thyroid ultrasound and antithyroxiperoxidase antibodies greater than 200 IU/ml. RESULTS: Females with occupational exposure to ionizing radiation had more often AITD than nonexposed females (10.0 vs. 3.4%; P < 0.05). This association persisted after adjustment for relevant confounders (odds ratio, 3.46; 95% confidence interval, 1.16-10.31; P < 0.05). In males, there were too few subjects who fulfilled the criteria of AITD, but the association between the exposure to radiation and hypoechogenicity of the thyroid gland barely missed statistical significance (odds ratio, 2.20; 95% confidence interval, 0.92-5.26; P = 0.08). In both females and males, subjects who reported a length of exposure of more than 5 yr exhibited the highest risk of the endpoints. CONCLUSIONS: We conclude that occupational exposure to ionizing radiation is related to the risk of AITD. The usage of thyroid protection shields by radiation workers is strongly recommended. PMID- 15886238 TI - Long-acting somatostatin analog therapy of acromegaly: a meta-analysis. AB - CONTEXT: Although considerable data exist on the use of long-acting somatostatin analogs to treat acromegaly, their reported efficacy differs substantially among trials. OBJECTIVE: We conducted a meta-analysis to derive definitive estimates of their efficacy for biochemical control and tumor shrinkage. DATA SOURCES: A search of literature was conducted through 2003, primarily via PubMed. STUDY SELECTION: Inclusion criteria, met in 44 trials, included at least 3 months of secondary octreotide long-acting release (LAR) or lanreotide slow release (SR) therapy or of primary octreotide LAR, lanreotide SR, or sc octreotide therapy and clearly reported data on biochemical efficacy and/or tumor shrinkage. Fifty other trials screened did not meet analysis inclusion criteria. DATA EXTRACTION: Data were extracted by three independent observers. DATA SYNTHESIS: Among subjects not selected for somatostatin analog responsiveness before study entry, both GH efficacy criteria and IGF-I normalization were met in a greater proportion of those treated with octreotide LAR vs. lanreotide SR (GH: B = 0.2310, P = 0.016; IGF-I: B = 0.2325, P = 0.007). Prestudy selection for somatostatin analog responsiveness was not a significant predictor of meeting GH efficacy criteria (B = 0.0992; P = 0.12). Preselection was a positive predictor of IGF-I normalization rate (B = 0.1213; P = 0.04), which was greater among preselected than unselected subjects (B = 0.1472; P = 0.0475). IGF-I normalization occurred in a greater proportion of secondary octreotide LAR- vs. primary octreotide-treated subjects (B = 0.2056; P = 0.009). The odds of tumor shrinkage more than 10% were lower in the unselected vs. preselected subjects. However, the effect of drug type was an important predictor of shrinkage; such that regardless of preselection or not, the odds of shrinkage with lanreotide SR were lower than with octreotide LAR (P = 0.003). Shrinkage greater than 10% occurred in a higher percentage of primary octreotide LAR-treated vs. primary octreotide sc-treated subjects (odds ratio = 9.4; P < 0.0001). The overall rate of tumor increase was 1.4%. CONCLUSIONS: In this meta-analysis, we have shown that the efficacy of octreotide LAR is greater than lanreotide SR among subjects unselected for prior somatostatin analog responsiveness. Preselection is a significant positive predictor of IGF-I normalization and is associated with increased odds of tumor shrinkage, which is also greatest with octreotide LAR. Biochemical efficacy is similar, but tumor shrinkage is greater when these drugs are given as primary vs. secondary therapy. PMID- 15886239 TI - Tissue transglutaminase at embryo-maternal interface. AB - CONTEXT: Tissue transglutaminase (tTG) has a high affinity for fibronectin (FN) and is a coreceptor of both beta1 and beta3 integrin subunits. Considering the notion that FN and integrins have critical roles during the implantation process, this study was undertaken to elucidate the expression pattern and the potential physiological function of tTG at the embryo-maternal interface. METHODS: The primary cultures of human placentas from 15 legal elective abortions at the first trimester of normal pregnancies and endometrial biopsies of 12 female patients in the midluteal phase as well as normal trophoblastic cell lines (CRL) were employed to address these issues using several approaches, such as scanning and transmission electron microscopies, immunostaining for light and electron microscopies, western blotting, and function assays using GRGDSP hexapeptide and an antibody against tTG. RESULTS: The results demonstrated tTG expression on uterine pinopodes and lamellipodia of extravillous trophoblasts. The colocalization of tTG with beta1 and beta3 integrins and its interaction with alpha(v)beta3 integrin and integrin-associated proteins at focal adhesions of the extravillous trophoblasts were illustrated in the results of immunofluorescence, immunoblot, and coimmunoprecipitation studies. Furthermore, function assays revealed that tTG mediated the adhesion and spread of the placental cells on intact FN-coated and 42- and 110-kDa FN fragment-coated wells. CONCLUSION: In conclusion, our findings demonstrated for the first time that tTG actively participates in adhesion events at the embryo-maternal interface through its interaction with FN, at least in part, by activating integrin-signaling pathways. PMID- 15886240 TI - Transient hypothyroxinemia in preterm infants: the role of cord sera thyroid hormone levels adjusted for prenatal and intrapartum factors. AB - CONTEXT: Transient hypothyroxinemia is common in infants less than 30 wk gestation and is associated with neurodevelopmental deficits; it has no consensus definition. We previously suggested that appropriate ranges for postnatal serum T4 values are at least cord levels corrected to an equivalent gestational age if the fetuses had remained in utero. OBJECTIVE: The study objective is to investigate the contribution of prenatal and intrapartum factors (n = 27) to the variations in cord levels of iodothyronines, T4-binding globulin, and TSH, and to provide an appropriate definition of transient hypothyroxinemia. DESIGN: The study design is a cohort study (n = 620) in 11 Scottish neonatal intensive care units. PATIENTS: Infants were delivered at 23- to 42-wk gestation and recruited between January 1998 and September 2001. RESULTS: Using -2 SD of adjusted T4 cord levels applied to postnatal d-7 values of equivalent gestational age, 14% of the 23- to 27-wk group, 1% of the 28- to 30-wk group, and 3% of the 31- to 34-wk group are hypothyroxinemic; using -1 SD, the respective figures are 41, 23, and 12%. CONCLUSIONS: In the absence of neurodevelopmental follow-up studies to quantify transient hypothyroxinemia, a pragmatic criterion is necessary for selection of extreme preterm infants into clinical trials of T4 supplementation. We suggest the use of serum T4 levels on postnatal d 7 that are below -1 SD of adjusted cord T4 levels of equivalent gestational age. This criterion avoids over recruitment of the more mature infants in whom universal T4 supplementation is detrimental to neurodevelopmental outcome, but still allows selection of the least mature entrants on whom universal T4 supplementation is beneficial. PMID- 15886241 TI - Adiponectin predicts insulin resistance but not endothelial function in young, healthy adolescents. AB - BACKGROUND: Adiponectin, an adipocyte-derived hormone found in lower concentration with greater adiposity, is suggested to reduce the risk of insulin resistance, atherosclerosis, and cardiovascular disease. We tested this hypothesis in a healthy, nonobese population. METHODS AND RESULTS: Brachial artery flow-mediated endothelial-dependent vasodilation and distensibility, measures of vascular function relevant to the early atherosclerotic process, were determined in 294 adolescents (aged 13-16 yr) using high-resolution vascular ultrasound. Fasting insulin concentration and the homeostasis model assessment of insulin resistance were used to estimate insulin resistance. Fat mass was measured by bioelectric impedance analysis; fasting serum adiponectin concentration by RIA; and lipid profile, fasting insulin, glucose, and C-reactive protein concentrations using standard laboratory techniques. Adiponectin concentration was associated with insulin resistance independent of potential confounding factors (e.g. -1.3% change in fasting insulin concentration per 10% increase in adiponectin concentration; 95% confidence interval, -2.4% to -0.1%; P = 0.03), but not with flow-mediated endothelial-dependent vasodilation or arterial distensibility. CONCLUSIONS: Lower adiponectin concentration was associated with lower insulin sensitivity in a healthy, nonobese population. Our study supports the hypothesis that adiponectin benefits insulin sensitivity from a young age but, in contrast to experimental models and data from older subjects, does not affect vascular changes associated with early atherosclerosis. PMID- 15886242 TI - Clinical and biochemical characteristics of adrenocorticotropin-secreting macroadenomas. AB - CONTEXT: Cushing's disease as a result of a pituitary macroadenoma is an uncommon cause of Cushing's syndrome, and reports in the published literature are few and of limited size. OBJECTIVE: Our objective was to establish the clinical and biochemical characteristics of macroadenomas associated with Cushing's disease compared with a large cohort of microadenomas and to assess their response to therapy. DESIGN: We conducted a retrospective case-records study for the years 1964-2001. SETTING: The study occurred at a tertiary referral hospital center. PATIENTS: Patients had Cushing's disease presenting with a pituitary macroadenoma, in comparison with a large group of microadenoma patients. INTERVENTIONS: Interventions included therapy with surgery and radiotherapy. MAIN OUTCOME MEASURES: Outcome measures included basal and dynamically responsive plasma ACTH and cortisol levels and response to treatment. RESULTS: We identified 18 patients with Cushing's disease secondary to a macroadenoma; basal 0900 h plasma ACTH was 135.8 +/- 32.5 and 45.0 +/- 4.3 ng/liter (mean +/- SEM), respectively, in macroadenomas and microadenomas (P = 0.013). Mean 0900 h serum cortisol was significantly increased in the macroadenomas (27.5 +/- 3.0 microg/dl, 759.6 +/- 82.6 nmol/liter, vs. 22.6 +/- 0.6 microg/dl, 624.7 +/- 16.4 nmol/liter) (P = 0.021). Testing with high-dose dexamethasone showed less suppression in the macroadenomas (57.6 +/- 8.7% vs. 74.4 +/- 2.1%; P = 0.02) and an attenuated ACTH response to CRH. For all biochemical variables there was considerable overlap between the two groups. Few patients with macroadenomas were cured by surgery. CONCLUSIONS: Pituitary macroadenomas causing Cushing's disease have biochemical features largely distinct from patients harboring microadenomas but represent one end of a continuum. PMID- 15886243 TI - Increasing serum betahydroxybutyrate concentrations during the 72-hour fast: evidence against hyperinsulinemic hypoglycemia. AB - OBJECTIVE: We have determined whether the behavior of betahydroxybutyrate (BOHB) during the 72-h fast of persons without evidence of hyperinsulinemic or any form of organic hypoglycemia might provide indicators of a negative fast. DESIGN: Twenty-one patients with surgically confirmed insulinoma and 34 patients with negative 72-h fasts had BOHB measured every 6 h until Whipple's triad in the former and until 72-h in the latter. RESULTS: Quadratic regression curves of BOHB from the negative fasts showed the typical curve to be flat initially, then increase in a manner that was roughly linear. Using time-specific medians, the changes were: 12-18 h, 0% increase; 18-36 h, 333% increase; 36-54 h, 210% increase, and 54-72 h, 167% increase. In contrast, patients with insulinoma had suppressed BOHB concentrations. Two successive BOHB values in excess of the 18-h level seemed to portend a negative fast. By using the previously published criterion of BOHB more than 2.7 mmol/liter (a surrogate for hypoinsulinemia and thereby an indicator of a negative fast), 74% of persons with a negative fast reached this level before the 72-h point. CONCLUSIONS: Serial measurements of BOHB during the 72-h fast have the potential to provide not only clues during the fast that it will ultimately be negative, but also the opportunity to truncate the fast if the endpoint BOHB criterion for a negative fast is met before 72 h. PMID- 15886244 TI - Sex-related differences in stimulated hypothalamic-pituitary-adrenal axis during induced gonadal suppression. AB - CONTEXT: Sex-related differences in the stress response are well described in the animal literature but in humans are inconsistent and appear to reflect both the method used to stimulate the hypothalamic-pituitary-adrenal (HPA) axis and the age of the subjects. Sex-related differences in reproductive steroid levels further confound efforts to define the specific role of the sex of the individual in stress axis responsivity. OBJECTIVE: The aim of this study was to address this role independent of differences in reproductive steroid levels. We compared HPA axis response to pharmacological (CRH) and physiological (exercise) stressors in two groups of young to middle-aged (18-45 yr) men (n = 10 and 8) and women (n = 12 and 13) undergoing gonadal suppression with leuprolide acetate (monthly im injection of 7.5 mg in men and 3.75 mg in women). DESIGN: Exercise and CRH stimulation tests were performed during induced hypogonadal conditions. SETTING: The study was conducted at a National Institutes of Health Clinical Center Outpatient Clinic. PATIENT OR OTHER PARTICIPANTS: Male and female normal volunteers participated in the study. MAIN OUTCOME MEASURES: The main outcome measures were stimulated ACTH and cortisol levels. RESULTS: Both CRH (1 microg/kg) stimulation and graded treadmill exercise stimulation occurred in the month after the second leuprolide injection to ensure gonadal suppression. Despite the absence of sex differences in estradiol or testosterone at the time of testing, men showed increased stimulated ACTH (repeated-measures ANOVA for CRH, P < 0.005) and cortisol (repeated-measures ANOVA for exercise, P < 0.05) compared with women. Among the summary measures, area under the curve (AUC) for cortisol was significantly greater in men than women after exercise. Although the AUC for ACTH was not significantly different across sexes, the initial AUC (0-30 min) was significantly greater in men for both procedures. No significant sex differences were found in a measure of adrenal responsivity, the cortisol to ACTH ratio, for either procedure. Cortisol-binding globulin levels did not differ between men and women and were not correlated with stimulated HPA axis measures. These data confirm earlier reports of sex differences in stimulated HPA axis activity and demonstrate that these differences exist even under induced hypogonadal conditions (i.e. in the absence of characteristic differences in reproductive steroids). PMID- 15886245 TI - Vildagliptin, a dipeptidyl peptidase-IV inhibitor, improves model-assessed beta cell function in patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: The dipeptidyl peptidase IV inhibitor, vildagliptin, increases levels of intact glucagon-like peptide-1 (GLP-1) and improves glycemic control in patients with type 2 diabetes. Although GLP-1 is known to stimulate insulin secretion, vildagliptin does not affect plasma insulin levels in diabetic patients, suggesting that more sophisticated measures are necessary to ascertain the influence of vildagliptin on beta-cell function. METHODS: This study examined the effects of 28-d treatment with vildagliptin (100 mg, twice daily; n = 9) vs. placebo (n = 11) on beta-cell function in diabetic patients using a mathematical model that describes the insulin secretory rate as a function of glucose levels (beta-cell dose response), the change in glucose with time (derivative component), and a potentiation factor, which is a function of time and may reflect the actions of nonglucose secretagogues and other factors. RESULTS: Vildagliptin significantly increased the insulin secretory rate at 7 mmol/liter glucose (secretory tone), calculated from the dose response; the difference in least squares mean (deltaLSM) was 101 +/- 51 pmol.min(-1).m(-2) (P = 0.002). The slope of the beta-cell dose response, the derivative component, and the potentiation factor were not affected. Vildagliptin also significantly decreased mean prandial glucose (deltaLSM, -1.2 +/- 0.4 mmol/liter; P = 0.01) and glucagon (deltaLSM, -10.7 +/- 4.8 ng/liter; P = 0.03) levels and increased plasma levels of intact GLP-1 (deltaLSM, +10.8 +/- 1.6 pmol/liter; P < 0.0001) and gastric inhibitory polypeptide (deltaLSM, +43.4 +/- 9.4 pmol/liter; P < 0.0001) relative to placebo. CONCLUSION: Vildagliptin is an incretin degradation inhibitor that improves beta-cell function in diabetic patients by increasing the insulin secretory tone. PMID- 15886246 TI - Transfection of antisense chorionic gonadotropin beta gene into choriocarcinoma cells suppresses the cell proliferation and induces apoptosis. AB - CONTEXT: Choriocarcinoma cells not only synthesize human chorionic gonadotropin (hCG), but also express LH/CG receptors on the cell membrane. This suggests that the hCG and LH/CG receptors may play a role in regulating the biological function of choriocarcinoma cells in an autocrine/paracrine manner. OBJECTIVE AND METHODS: The objective of this study was to ascertain whether the inhibition of CGbeta gene expression in choriocarcinoma cells affects their proliferation and apoptosis. Expression vector bearing antisense CGbeta gene was transfected into the choriocarcinoma cell line, JAr. CGbeta protein synthesis was monitored by Western immunoblot, and CGbeta mRNA expression was determined by RT-PCR. Cell proliferation was assessed by 3-[4,5-dimethlthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and nuclear incorporation of 5-bromo-2'-deoxyuridine, and the apoptosis-positive rate was assessed by terminal deoxynucleotidyltransferase mediated deoxy-UTP nick end labeling analysis and nuclear staining with Hoechst 32258. RESULTS: JAr cells transfected with antisense CGbeta gene (JAr-aCGbeta cells) showed a significant decrease in hCG production and cell proliferation compared with untransfected and mock-transfected cells. The apoptosis-positive rate of the JAr-aCGbeta cells significantly increased compared with that of the controls. LH/CG receptor expression in JAr-aCGbeta cells decreased compared with that in controls. By contrast, supplementation of exogenous hCG significantly increased the LH/CG receptor expression and viability of JAr-aCGbeta cells. CONCLUSIONS: These results suggest that hCG, through its binding to the LH/CG receptor, may augment proliferation and inhibit apoptosis in choriocarcinoma JAr cells, and that the introduction of an antisense gene may be a potential approach to the inhibition of choriocarcinoma cell growth. PMID- 15886247 TI - Metformin and weight loss in obese women with polycystic ovary syndrome: comparison of doses. AB - CONTEXT: Metformin treatment of women with polycystic ovary syndrome (PCOS) is widespread, as determined by studies with diverse patient populations. No comparative examination of weight changes or metabolite responses to different doses has been reported. OBJECTIVE: The aim of this study was to determine whether different doses of metformin (1500 or 2550 mg/d) would have different effects on body weight, circulating hormones, markers of inflammation, and lipid profiles. DESIGN: The study included prospective cohorts randomized to two doses of metformin. SETTING: The study was performed at a university teaching hospital with patients from gynecology/endocrinology clinics. PATIENTS: The patients studied were obese (body mass index, 30 to <37 kg/m2; n = 42) and morbidly obese (body mass index, > or =37 kg/m2; n = 41) women with PCOS. INTERVENTION: Patients were randomized to two doses of metformin, and parameters were assessed after 4 and 8 months. MAIN OUTCOME MEASURES: The main outcome measures were changes in body mass, circulating hormones, markers of inflammation, and lipid profiles. RESULTS: Intention to treat analyses showed significant weight loss in both dose groups. Only the obese subgroup showed a dose relationship (1.5 and 3.6 kg in 1500- and 2550-mg groups, respectively; P = 0.04). The morbidly obese group showed similar reductions (3.9 and 3.8 kg) in both groups. Suppression of androstenedione was significant with both metformin doses, but there was no clear dose relationship. Generally, beneficial changes in lipid profiles were not related to dose. CONCLUSION: Weight loss is a feature of protracted metformin therapy in obese women with PCOS, with greater weight reduction potentially achievable with higher doses. Additional studies are required to determine whether other aspects of the disorder may benefit from the higher dose of metformin. PMID- 15886248 TI - A common single nucleotide polymorphism in exon 10 of the human follicle stimulating hormone receptor is a major determinant of length and hormonal dynamics of the menstrual cycle. AB - CONTEXT: FSH is essential for follicular maturation. Data from ovarian hyperstimulation cycles suggest that FSH action is attenuated by a frequent single nucleotide polymorphism of the FSH receptor gene exchanging Asn for Ser at codon 680. OBJECTIVE: We hypothesized that the FSH receptor genotype influences menstrual cycle dynamics. DESIGN: Menstrual cycle was monitored from the midluteal phase through ovulation until the consecutive menstruation. SETTING: The study was conducted at the University research center. SUBJECTS: Women homozygous for the Asn680 (n = 12) and Ser680 (n = 9) variants with normal menstrual cycles volunteered for the study. INTERVENTIONS: There were no interventions. MAIN OUTCOME MEASUREMENTS: Follicular growth, serum LH, FSH, estradiol, progesterone, inhibin A, inhibin B and antimullerian hormone were measured. RESULTS: During the luteo-follicular transition, serum levels of estradiol, progesterone, and inhibin A were significantly lower, and FSH started to rise earlier in the Ser680/Ser680 group. FSH levels were steadily and significantly higher, and the mean area under the FSH curve was 31% greater in this group (P < 0.002). No differences were observed in estradiol, inhibin B, and growth velocities of dominant follicles. The time from luteolysis to ovulation was significantly longer in women with the Ser680/Ser680 (13.6 +/- 1.01 d) compared with Asn680/Asn680 (11.3 +/- 0.61 d, P < 0.05) genotype with a significant difference in total menstrual cycle length (29.3 vs. 27.0 d, respectively; P < 0.05). CONCLUSIONS: The FSH receptor Ser680/Ser680 genotype is associated with higher ovarian threshold to FSH, decreased negative feedback of luteal secretion to the pituitary during the intercycle transition, and longer menstrual cycles. PMID- 15886249 TI - Inhibition of human trophoblast invasiveness by high glucose concentrations. AB - CONTEXT: Trophoblast invasion of the uterus is regulated by local microenvironmental factors. OBJECTIVE: Because certain conditions may affect uterine glucose levels during placentation, the aim of this study was to determine the effect of glucose concentration on trophoblast invasion. RESULTS: Compared with incubation in 0.2 and 2.5 mm glucose, a 24-h incubation in increasing glucose concentrations (5 and 10 mm) resulted in up to a 62% inhibition (P < 0.01) of the in vitro invasiveness of immortalized HTR-8/SVneo trophoblasts. This decreased invasiveness in 5 and 10 mm glucose was paralleled by inhibition of a plasminogen activator (PA) activity corresponding to active urokinase-type PA (uPA). Inhibition of pro-uPA binding to the uPA receptor decreased the invasiveness of cells incubated in 0.2 and 2.5 mm glucose to levels observed in cells incubated in higher glucose concentrations (P < 0.05). Gelatin zymography and Western blot analysis revealed that the levels of matrix metalloproteinase-2 and -9, PA inhibitor-1, and uPA receptor were unaffected by glucose. Glucose transporter-1 levels were 26 and 34% higher in cells cultured in 2.5 and 0.2 mm glucose, respectively, vs. 5 or 10 mm glucose (P < 0.05). In contrast, glucose transporter-3 levels were not affected by incubation in various glucose concentrations. CONCLUSIONS: These findings indicate that high glucose concentrations inhibit the invasiveness of HTR-8/SVneo cells by preventing uPA activation. Therefore, through its effects on uPA activity, glucose may be an important regulator of trophoblast invasiveness during implantation and placentation. PMID- 15886250 TI - Gene expression profiling of orbital adipose tissue from patients with Graves' ophthalmopathy: a potential role for secreted frizzled-related protein-1 in orbital adipogenesis. AB - CONTEXT: The signs and symptoms of Graves' ophthalmopathy (GO) result from inflammation and increased volume of the orbital adipose tissues and extraocular muscles. OBJECTIVE: Our objective was to identify differentially regulated genes that may be involved in stimulating the orbital adipose tissue expansion seen in GO. DESIGN: Gene expression profiling was used to compare genes expressed in orbital adipose tissues from GO patients and normal individuals. SETTING: The study took place at a private practice tertiary referral center. PATIENTS: Orbital adipose tissues were collected at transantral orbital decompression surgery from 20 euthyroid patients undergoing this procedure for severe GO and at early autopsy from eight normal individuals having no evidence of thyroid or ocular disease. RESULTS: Of the 12,686 genes analyzed, 25 known genes were increased in expression (>4-fold) in GO orbital tissues, whereas 11 genes were decreased (>4-fold). Up-regulated genes, confirmed by quantitative RT-PCR, included secreted frizzled-related protein-1 (sFRP-1; 18.5-fold) and several adipocyte-related genes, including peroxisome proliferator activated receptor gamma (44.1-fold) and adiponectin (25-fold). Treatment in vitro of GO orbital preadipocytes with recombinant sFRP-1 (100 nm) significantly increased adiponectin (2.0-fold; P < 0.05), leptin (7-fold; P < 0.002), and TSH receptor mRNA (13-fold; P < 0.003) levels and enhanced Oil red-O staining in the cultures. CONCLUSIONS: These results support the concept that orbital adipogenesis is enhanced in GO and suggest that elevated levels of sFRP-1 in the GO orbit may be involved in stimulating this pathogenic process. PMID- 15886251 TI - Resistin stimulation of 17alpha-hydroxylase activity in ovarian theca cells in vitro: relevance to polycystic ovary syndrome. AB - CONTEXT: A newly discovered hormone resistin has been shown to be increased in women with polycystic ovary syndrome (PCOS). OBJECTIVE: The purpose of this study was to confirm increased resistin concentrations in women with PCOS and to test the direct effect of resistin on human theca cell androgen production. DESIGN: Resistin was measured in fasting serum samples by RIA. To test the direct effects of resistin on ovarian androgen biosynthesis, human theca cells were cultured with resistin for 3 d in the presence and absence of forskolin and insulin. PATIENTS: Fasting serum samples were obtained from 45 women with PCOS and 74 regularly cycling premenopausal control women in the follicular phase of their menstrual cycles, and ovarian theca cell cultures were established from two control women. RESULTS: The mean serum resistin concentration was increased (40%) in women with PCOS. Serum resistin concentrations correlated positively with body mass index and testosterone in PCOS women but not in controls. There were no significant correlations between resistin and fasting insulin or indicators of insulin resistance when corrected for body mass index. In cultured human theca cells, basal 17alpha-hydroxylase activity was unchanged by resistin alone, but resistin enhanced 17alpha-hydroxylase activity in the presence of forskolin or a combination of forskolin plus insulin. Resistin (> or =1 ng/ml) augmented forskolin and forskolin plus insulin stimulation of CYP17 mRNA expression in a concentration-dependent manner. CONCLUSION: These data indicate that abnormal resistin secretion in PCOS may play a role in causing ovarian hyperandrogenism. PMID- 15886252 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin increases glycodelin gene and protein expression in human endometrium. AB - CONTEXT: Glycodelin (GdA) is an immunosuppressive endometrial glycoprotein critical for embryonic implantation and pregnancy establishment. OBJECTIVE: The aim of the present study was to examine the effect of dioxin [2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD)] on GdA production in human endometrial cells. DESIGN: Controlled endometrial explant (EE) and cell cultures were used in this study. SETTING: Work was conducted at university hospital research laboratories in Bern, Switzerland, and in San Francisco, California. PATIENTS: Ovulatory women provided endometrial biopsies in the proliferative or secretory phase. INTERVENTION(S): EEs and cells were cultured without and with TCDD. MAIN OUTCOME MEASURE(S): GdA protein and gene expression were quantified. RESULTS: A 2.5-fold increase in GdA production was demonstrated in EEs treated with 10 nm TCDD for 9 d. Fluorography revealed a 3- to 4-fold increase in new GdA biosynthesis and secretion in TCDD-treated endometrial epithelial cells. Because the action of dioxin is mediated by the aryl hydrocarbon receptor (AhR), we ascertained that primary epithelial and Ishikawa cells express AhR. Dose responses to TCDD and expressed AhR were established in transiently transfected Ishikawa cells using luciferase fusion vectors containing 1.0 kb of 5' flanking DNA relative to the GdA transcriptional start site but not when shorter promoter constructs were used. A dioxin response element was mapped to nucleotides -539 to -533 of the gene promoter and verified by site-directed mutagenesis. CONCLUSIONS: We demonstrated a direct AhR-mediated effect of dioxin on GdA gene transcription and protein secretion that might influence human female fertility. PMID- 15886253 TI - Soluble fms-like tyrosine kinase 1 is increased in preeclampsia but not in normotensive pregnancies with small-for-gestational-age neonates: relationship to circulating placental growth factor. AB - CONTEXT: An excess of the soluble receptor, fms-like tyrosine kinase 1 (sFlt-1) may contribute to maternal vascular dysfunction in women with preeclampsia by binding and thereby reducing concentrations of free vascular endothelial growth factor and placental growth factor (PlGF) in the circulation. The putative stimulus for increased sFlt-1 during preeclampsia, placental hypoxia due to poor perfusion, is common to both preeclampsia and idiopathic intrauterine growth restriction. However, the latter condition occurs without maternal vascular disease. OBJECTIVE: We asked whether, as with preeclampsia, sFlt-1 is increased and free PlGF is decreased in villous placenta and maternal serum of normotensive women with small-for-gestational-age (SGA) neonates. STUDY DESIGN: This was a case-control study using banked samples. Groups of women with SGA neonates (birth weight centile < 10th) and women with preeclampsia were matched to separate sets of normal pregnancy controls based on gestational age at blood sampling (serum) or gestational age at delivery (placenta). RESULTS: sFlt-1 levels were higher in preeclamptics than controls (serum, P < 0.0001; placental protein, P = 0.03; placental mRNA, P = 0.007) but not increased in SGA pregnancies. PlGF was lower in both preeclampsia (serum, P < 0.0001; placental protein, P = 0.05) and SGA (serum, P = 0.0008; placental protein, P = 0.03) compared with their controls. PlGF in preeclampsia and SGA groups did not differ. CONCLUSIONS: These data are consistent with a role for sFlt-1 in the maternal manifestations of preeclampsia. In contrast to preeclampsia, sFlt-1 does not appear to contribute substantially to decreased circulating free PlGF in SGA pregnancies in the absence of a maternal syndrome. PMID- 15886254 TI - Differences in serum sex hormone and plasma lipid levels in Caucasian and African American premenopausal women. AB - CONTEXT: Risk of coronary heart disease is higher in African-American than in Caucasian women. OBJECTIVE: The aim of this study was to evaluate the contribution of sex hormone levels, race, and measures of body fat to the variation in plasma lipid levels, a well-established risk factor for coronary heart disease. DESIGN: This was a cross-sectional study. SETTING: The study was conducted in the general community. STUDY PARTICIPANTS: Sixty Caucasian and 117 African-American premenopausal women participated. MAIN OUTCOME MEASURES: Body weight, body mass index (BMI), and waist to hip circumference ratio (WHR), as well as plasma lipid and serum sex hormone levels, were assessed. RESULTS: Relative to Caucasian women, African-American women had significantly higher mean BMI (23.92 +/- 3.87 vs. 26.99 +/- 5.87 kg/m2, respectively; P < 0.001), and WHR (0.733 +/- 0.052 vs. 0.757 +/- 0.068; P < 0.03). Also, plasma triglyceride (TG) levels were significantly lower in African-American women (81 +/- 61 vs. 55 +/- 24 mg/dl; P < 0.0001). Serum estrone sulfate (556 +/- 323 vs. 442 +/- 332 pg/ml, Caucasian vs. African-American; P < 0.001), estradiol (E2) (55.1 +/- 43.6 vs. 35.8 +/- 17.7 pg/ml; P < 0.0001), androstenedione (2.6 +/- 0.9 vs. 1.6 +/- 0.7 ng/ml; P < 0.0001), and testosterone (0.36 +/- 0.12 vs. 0.31 +/- 0.19 ng/ml; P < 0.002) levels were significantly lower in African-American women than in Caucasian women. After correction for the effects of age, BMI, and WHR, serum E2 levels were significantly and positively associated with plasma high-density lipoprotein cholesterol levels in all women, and serum estrone sulfate levels with plasma total cholesterol and TG levels in African-American women. CONCLUSIONS: Our results indicate that race is an important determinant of plasma TG and serum sex hormone levels, even after adjustment for differences in body size. A significant association between endogenous E2 and high-density lipoprotein cholesterol levels exists in premenopausal women, independent of their race. PMID- 15886255 TI - Lower-body adiposity and metabolic protection in postmenopausal women. AB - CONTEXT: It has been suggested that the propensity to store fat in the gluteal femoral region may be cardioprotective. OBJECTIVE: The primary aim of this study was to test whether the favorable associations of leg fat with risk factors for cardiovascular disease persist after controlling for the highly unfavorable effects of abdominal (visceral or sc) adiposity in postmenopausal women. STUDY PARTICIPANTS: The study included 95 postmenopausal women [age, 60 +/- 8 yr (mean +/- SD)]. MAIN OUTCOMES: Whole-body and regional fat distribution was measured using dual-energy x-ray absorptiometry and abdominal computed tomography. Markers of insulin resistance and dyslipidemia were determined from oral glucose tolerance tests and fasted lipid and lipoprotein measurements, respectively. Primary outcomes were: fasting insulin (INS0), area under the insulin curve (INS(AUC)), product of the oral glucose tolerance test insulin and glucose AUC (INS(AUC) - GLU(AUC)), serum triglycerides (TG), and high-density lipoprotein (HDL) cholesterol. RESULTS: Controlling for trunk fat revealed a favorable effect of leg fat on INS0, INS(AUC), INS(AUC) x GLU(AUC), TG, and HDL. However, after controlling for either visceral or sc abdominal adiposity, TG was the only risk factor for which the favorable effect of leg fat persisted. CONCLUSIONS: The lack of an association between leg fat and most of the risk factors, after adjusting for abdominal visceral or sc fat, suggests an overriding deleterious influence of abdominal adiposity on cardiovascular risk. Nevertheless, our finding that regional adipose tissue depots have apparent independent and opposing effects on serum TG supports the need for further research into the physiological mechanisms governing these effects. PMID- 15886256 TI - A nationwide survey of mortality in acromegaly. AB - CONTEXT: Increased mortality in acromegaly has been confined to those with posttreatment basal GH of 2.5 microg/liter or greater, but the impact of IGF-I and pituitary radiotherapy on mortality has remained controversial. OBJECTIVE: The purpose of this nationwide survey was to examine the all-cause mortality of patients with acromegaly and evaluate the impact of treatment outcome and mode of treatment on survival. DESIGN, SETTING, AND PATIENTS: All-cause mortality of all patients with acromegaly diagnosed during January 1980 and December 1999 in the five university hospitals of Finland was followed up by the end of 2002 (12.5 +/- 5.6 yr) and compared with that of the general population by using age- and gender adjusted standardized mortality ratios (SMRs). Logistic regression analysis was used to investigate factors related to mortality within the survey population. MAIN OUTCOME MEASURE: Mortality was the main outcome measure. RESULTS: Of the 334 patients, 56 (16.8%) had died during follow-up. SMR of the patients was 1.16 [confidence interval (CI) 0.85-1.54, not significant (NS)]. However, patients with basal serum GH concentration 2.5 microg/liter or greater (SMR 1.63, CI 1.10 2.35, P < 0.001) measured 5.2 +/- 4.4 yr after the initial treatment, and those irradiated (SMR 1.69, CI 1.05-2.58, P < 0.001) showed excess mortality. In a multivariate model, the effect of radiotherapy was of borderline significance only (P = 0.083). Posttreatment IGF-I levels, available for 72.2% of the patients, did not have impact on mortality. CONCLUSIONS: The posttreatment basal GH concentration less than 2.5 microg/liter in acromegalic patients is associated with a normal lifespan. Excess mortality is confined to poorly controlled patients and possibly those who have received conventional radiotherapy. PMID- 15886257 TI - Peroxisome proliferator-activated receptor-gamma agonists suppress adrenocortical tumor cell proliferation and induce differentiation. AB - CONTEXT: Thiazolidinediones (TZDs) have been implemented into clinical practice for the treatment of type 2 diabetes mellitus as specific peroxisome proliferator activated receptor (PPAR)-gamma ligands. Moreover, recent evidence has suggested that TZDs might have favorable effects in the treatment of a variety of tumors as differentiation-inducing agents. Adrenocortical carcinoma (ACC) is a rare tumor entity with poor prognosis due to its highly malignant phenotype and lack of effective treatment options. OBJECTIVE: The purpose of this study was to investigate effects of TZDs on adrenocortical cancer cells. RESULTS: PPARgamma mRNA expression was detectable in all adrenocortical tumors including ACCs at similar levels. Furthermore, incubation of the adrenocortical tumor cell line NCI h295 with the PPARgamma agonist rosiglitazone led to a decrease in cell viability, a decrease of cellular proliferation, and an increase in apoptosis as well as steroidogenesis. On the molecular level, NCI h295 cells expressed higher levels of ACTH receptor (melanocortin receptor-2) mRNA upon treatment, whereas cyclin E mRNA was reduced, thus reflecting a shift toward an expression pattern found in less aggressive adrenocortical tumors in vivo. Accordingly, luciferase experiments confirmed an increased promoter activity for the melanocortin receptor-2 after stimulation with rosiglitazone. Coincubation with the specific PPARgamma antagonist GW9662 demonstrated the inhibition of TZD-induced increase in steroidogenesis, whereas growth suppression upon TZD treatment was not affected by GW9662. CONCLUSIONS: Thus, both PPARgamma-dependent and PPARgamma independent effects of TZD treatment are likely to contribute to the observed phenotypical effects on NCI h295 cells. Taken together, these data indicate that TZDs might have the potential to become an additional treatment option as differentiation-inducing agents in patients with ACC. PMID- 15886258 TI - The genome stability network. PMID- 15886260 TI - Volume guarantee ventilation, interrupted expiration, and expiratory braking. AB - BACKGROUND: In volume guarantee ventilation with the Drager Babylog 8000 ventilator, inspiratory and expiratory flows are monitored and the expiratory tidal volume calculated following each inflation. The pressure for the next inflation is modified to ensure the expired tidal volume is close to the set value. AIM: To investigate interrupted expiration observed during volume guarantee ventilation of spontaneously breathing, ventilated infants. METHODS: Spontaneously breathing infants, ventilated with volume guarantee, had recordings of gas flow, ventilator pressures, tidal volume waveforms, oximetry, heart rate, and transcutaneous oxygen and carbon dioxide during 10 minute recordings. RESULTS: A total of 6540 inflations were analysed from 10 infants; 62% were triggered. Two different patterns were found: (1) Normal volume guarantee pattern with 97% of triggered and 91% untriggered inflations. It had a normal expiratory curve and a mean expired tidal volume within 3% of the set volume, but a large variation due to the babies' breathing. (2) A pattern of interrupted expiratory flow after approximately 3% of inflations due to a small inspiration (approximately 1.3 ml/kg) during expiration. This led the ventilator to calculate an inappropriate total expired tidal volume for that inflation and an increase in the pressure for the next inflation. CONCLUSIONS: After about 3% of inflations, with volume guarantee ventilation, interruption of the expiration causes an increased pressure for the next inflation of approximately 4.9 cm H2O, compared with normal volume guarantee inflation. The interrupted expiration is most likely to be due to diaphragmatic braking. PMID- 15886261 TI - Prevalence of autistic spectrum disorders in Lothian, Scotland: an estimate using the "capture-recapture" technique. PMID- 15886262 TI - Is it necessary to change the classification of {beta}-lactamases? PMID- 15886263 TI - Trimethoprim/sulfamethoxazole resistance in clinical isolates of Burkholderia pseudomallei. AB - OBJECTIVES: Trimethoprim/sulfamethoxazole is commonly used to treat melioidosis. Antimicrobial susceptibility testing using the disc diffusion method is commonly used in melioidosis-endemic areas, but may overestimate resistance to trimethoprim/sulfamethoxazole. PATIENTS AND METHODS: We performed disc diffusion and Etest on isolates from the first positive culture for all patients presenting to Sappasithiprasong Hospital, Ubon Ratchathani, Thailand, with culture-confirmed melioidosis between 1992 and 2003. RESULTS: The estimated resistance rate for 1976 clinical Burkholderia pseudomallei isolates was 13% by Etest and 71% by disc diffusion. All isolates classed as either susceptible (n=358) or as having intermediate resistance (n=218) on disc diffusion were susceptible by Etest. Only 258 of the 1400 (18%) isolates classed as resistant on disc diffusion were resistant by Etest. CONCLUSIONS: Disc diffusion testing of B. pseudomallei may be useful as a limited screening tool in resource poor settings. Isolates assigned as 'susceptible' or 'intermediate' by disc diffusion may be viewed as 'susceptible'; those assigned as 'resistant' require further evaluation by MIC methodology. PMID- 15886264 TI - Role of viral kinetics under HCV therapy in HIV/HCV-coinfected patients. AB - Patients coinfected with hepatitis C virus (HCV) and human immunodeficiency virus (HIV) are less responsive to anti-HCV therapies and are at a higher risk of toxicity than HCV monoinfected patients. HCV viral kinetics is the basis for the study of response to interferon-based therapy and for predicting sustained virological response (SVR). A lack of early virological response (EVR; undetectable HCV RNA or a decrease of >/=2 log(10) from baseline) after 12 weeks of pegylated interferon (peg-IFN) plus ribavirin (RBV) is an equally reliable predictor of lack of SVR in HIV/HCV-coinfected patients and in the monoinfected HCV population. Early stopping rules are particularly important in coinfected HIV/HCV patients, considering their low chances of response in the more difficult to-treat HCV genotypes 1 and 4 (<30%). Several factors have been involved in this low efficacy, including higher baseline HCV viraemia, slower viral kinetics decay under interferon pressure and a defective immune substratum. A better understanding of HCV viral kinetics under HCV therapy may be the basis for assaying different peg-IFN plus RBV schedules, such as induction or extending strategies, and may help physicians to make tailored decisions for the management of their patients. PMID- 15886265 TI - Diversity among 2481 Escherichia coli from women with community-acquired lower urinary tract infections in 17 countries. AB - OBJECTIVES: In the recently published ECO.SENS survey, the antimicrobial susceptibility of Escherichia coli from urinary tract infections in women in 16 European countries and Canada was investigated. This study reports the diversity among these E. coli. METHODS: The 2481 E. coli, typed with the PhenePlate (PhP) System utilizing the dynamics and end result of 11 biochemical reactions in a microplate system, were clustered and the Simpson's index of diversity calculated. RESULTS: Seventy-four Common PhP Types (CT) comprising 2067 isolates and 414 Single Types (Si) were identified. Of these, 916 isolates (37%) belonged to one of the four most frequent CT (arbitrarily numbered CT48, 10, 26 and 20). CT48 with 400 isolates and 11 different susceptibility patterns, was widely disseminated across Europe and Canada and was the most frequent type in 13 countries and the second most frequent in the remaining four countries. Sixty four per cent of the E. coli were susceptible to all eight investigated antimicrobials (CT48: 73%, CT10: 77%, CT26: 62% and CT20: 37%). Forty-six different susceptibility patterns were seen, the three most common being isolated resistance to ampicillin, resistance to ampicillin and trimethoprim, and isolated resistance to trimethoprim. Multiresistance, here defined as resistance to four or more of the investigated antibiotics, was distributed among E. coli belonging to several PhP types. CONCLUSIONS: There was no obvious correlation between the phenotypes identified with the PhP System and the susceptibility pattern. The data did not indicate clonal dissemination within or between countries as a major reason for differences in antimicrobial resistance rates. PMID- 15886266 TI - Susceptibilities of periodontopathogenic and cariogenic bacteria to antibacterial peptides, {beta}-defensins and LL37, produced by human epithelial cells. AB - OBJECTIVES: Antimicrobial peptides are one of the factors involved in innate immunity. The susceptibility of periodontopathogenic and cariogenic bacteria to the major antimicrobial peptides produced by epithelia was investigated. METHODS: Synthetic antimicrobial peptides of human beta-defensin-1 (hBD1), hBD2, hBD3 and LL37 (CAP18) were evaluated for their antimicrobial activity against oral bacteria. They included Actinobacillus actinomycetemcomitans (20 strains), Porphyromonas gingivalis (6), Prevotella intermedia (7), Fusobacterium nucleatum (7), Streptococcus mutans (5), Streptococcus sobrinus (5), Streptococcus salivarius (5), Streptococcus sanguis (4), Streptococcus mitis (2) and Lactobacillus casei (1). RESULTS: Although the four peptides had bactericidal activity against all bacteria tested, the degree of antibacterial activity was variable against the different strains and species. The antibacterial activity of hBD1 was lower than that of the other peptides. Among the bacteria tested in this study, F. nucleatum was highly susceptible to hBD3 and LL37, and S. mutans was highly susceptible to hBD3. We measured the Zeta-potential, representing the net charge of whole bacteria, to study the relationship between susceptibility to cationic peptide and the net charge of the bacteria. Although we found some correlation in A. actinomycetemcomitans strains, we did not find a definite correlation with all the bacterial species. CONCLUSIONS: These results indicate that beta-defensins and LL37 have versatile antibacterial activity against oral bacteria. PMID- 15886267 TI - The usefulness of MRI and PET imaging for the detection of parametrial involvement and lymph node metastasis in patients with cervical cancer. AB - OBJECTIVE: The purpose of this study is to elucidate the usefulness of magnetic resonance imaging (MRI) and positron emission tomography (PET) for the detection of parametrial involvement and lymph node metastasis in patients with cervical cancer. METHODS: Thirty-six patients with cervical cancer were retrospectively enrolled. MRI and PET scans were performed for all patients within a week before radical surgery. The criterion for malignancy on MRI was >1 cm short axis diameter of the suspected lymph node. On PET, only fluorodeoxyglucose (FDG) uptake was significantly higher than the background and, if this FDG uptake showed on at least two consecutive axial slices, then the lesion was considered as a malignancy. We compared the extent of tumor on the surgical findings with the FIGO staging, MRI and PET scans. RESULTS: The accuracy of FIGO and MRI staging was 67 and 84.4%, respectively. The accuracy for detecting pelvic lymph node metastasis was better for PET than for MRI (78 versus 67%, respectively). All FDG uptake lymph nodes were confirmed as metastatic lymph nodes by pathological evaluation; this included five lymph nodes <1 cm in diameter. CONCLUSION: MRI provides an improved evaluation of local tumor extension, but PET is more useful for the evaluation of pelvic lymph nodes than MRI; however, PET still misses microscopic disease. Further studies are necessary to evaluate the usefulness of PET/computed tomography (CT) for the accuracy of the disease extension and the cost-effectiveness of MRI, PET or PET/CT in patients with cervical cancer. PMID- 15886268 TI - Predictor of response to salvage radiotherapy in patients with PSA recurrence after radical prostatectomy: the usefulness of PSA doubling time. AB - BACKGROUND: We assessed predictors of response to salvage radiotherapy (sRT) in patients with prostate-specific antigen (PSA) recurrence after radical prostatectomy. METHODS: A total of 21 patients receiving sRT for PSA recurrence without systemic progression after radical prostatectomy had medical records available for retrospective review. We defined sRT as external beam radiotherapy for patients with a continuous increase in PSA level > or =0.2 ng/ml after radical prostatectomy. Response was defined as achievement of a PSA nadir of < or =0.1 ng/ml. Various pre-treatment parameters were evaluated retrospectively. RESULTS: The median follow-up period after sRT was 38 months. Of the 21 patients, 15 were good responders (71%). The only predictive factor was PSA doubling time (PSADT). Age and PSA level at diagnosis, Gleason score and surgical margin status were not significant predictors of response. The median PSADT in responders was 6.2 months versus 1.9 months in non-responders (P = 0.019). The patients with a PSADT of > or =5 months were all responders. CONCLUSION: PSADT appears to be a good predictor of response to sRT. sRT was especially effective when PSADT was > or =5 months. PMID- 15886269 TI - Phase II study of paclitaxel and carboplatin in advanced gastric cancer previously treated with 5-fluorouracil and platinum. AB - BACKGROUND: The combination of paclitaxel and carboplatin has been used to treat patients with many types of tumor, including gastric cancer. We evaluated the efficacy and safety of this combination in advanced gastric cancer patients previously treated with 5-fluorouracil and platinum. METHODS: Patients who had pathologically been proven to have measurable lesions were treated with paclitaxel (200 mg/m(2) for 3 h) and carboplatin [area under the concentration time curve (AUC = 6)] on day 1 and in 21 day cycles. RESULTS: A partial response was achieved in 10 of 45 patients [22%, 95% confidence interval (CI), 10-34]. Of the 32 patients previously treated with cisplatin, four (13%) achieved partial response, whereas, of the 13 patients previously treated with heptaplatin, six (46%) achieved partial response. In all patients, the median time to progression was 14 weeks (95% CI, 10-18), and the median overall survival was 32 weeks (95% CI, 26-38). The most common grade 3/4 adverse events were neutropenia (40% of patients) and neuropathy (2.2%). Two patients developed neutropenic fever. However, there were no treatment-related deaths. CONCLUSIONS: Combination chemotherapy with paclitaxel and carboplatin is feasible in patients with advanced gastric cancer who were previously treated with 5-fluorouracil and platinum. PMID- 15886270 TI - CD10-positive stromal cells in gastric carcinoma: correlation with invasion and metastasis. AB - BACKGROUND: CD10 is a cell surface metalloproteinase expressed by a variety of normal cell types, including lymphoid precursor cells, germinal center B lymphocytes and some epithelial cells. Although accumulating data indicate that CD10 expression by stromal cells is involved in colorectal carcinogenesis and it is a novel prognostic factor in breast carcinoma, CD10-positive stromal cells and their correlation with invasion and metastasis have not been studied in gastric carcinoma. The aim of this study was to immunohistochemically investigate the rate of CD10 production in the stromal cells in our gastric carcinoma collection and clarify its correlation with invasion and metastasis. METHODS: One hundred and sixteen cases of gastric carcinoma were analyzed immunohistochemically using a monoclonal CD10 antibody (clone 56C6). RESULTS: The expression of CD10 by stromal cells was significantly higher in the primary gastric carcinomas than in normal and dysplasia mucosas (P = 0.014). More frequent expression of CD10 by stromal cells was detected in differentiated carcinoma than in undifferentiated carcinoma (P < 0.001). CD10 expression by the stromal cells was associated with depth of invasion and lymph node metastasis (P < 0.05). Stromal CD10 expression was lower in gastric carcinoma without vessel invasion than in those with vessel invasion (P = 0.001). However, no association was observed between stromal CD10 expression and TNM stage. In differentiated carcinoma, stromal CD10 expression was associated with the depth of invasion, lymph node metastasis, vessel invasion and TNM stage (P < 0.01). CONCLUSION: These results indicate that stromal cells expressing CD10 may play an important role in gastric carcinogenesis. CD10 expression by stromal cells seems to promote invasion and metastasis of differentiated gastric carcinoma. PMID- 15886271 TI - An accelerated radiotherapy scheme using a concomitant boost technique for the treatment of unresectable stage III non-small cell lung cancer. AB - BACKGROUND: We designed a phase II trial for evaluation of the efficacy and tolerability of an accelerated concomitant boost radiotherapy scheme for the treatment of the patients with non-small cell lung cancer (NSCLC). METHODS: Thirty patients with unresectable stage IIIA/IIIB NSCLC were prospectively enrolled in this protocol. All patients were scheduled to receive 15 fractions of conventional radiotherapy in doses of 1.8 Gy, to a total of 27 Gy. For the last 10 treatment days, an accelerated concomitant boost schedule was started that was composed of 1.8 Gy/fraction/day, 5 days/week to the large field and 1.8 Gy/fraction/day to the boost field 6 h apart, to a total dose of 63 Gy/35 fractions/5 weeks. RESULTS: Median follow-up time was 13 months (range, 5-50 months; 3-year overall, disease-free, loco-regional disease-free and metastasis free survivals were 23%, 19%, 19% and 23%, respectively). The most common acute toxicity was esophagitis in 31% of patients with the Radiation Therapy Oncology Group and the European Organization for Research and Treatment of Cancer (RTOG/EORTC) criteria grade 1, and in 54% with grade 2. Radiation pneumonitis developed in 16% of patients with RTOG/EORTC grade 1. Three-year actuarial rate of late pulmonary and skin-subcutaneous toxicity were 12% and 16%, respectively. No late radiotherapy complications of spinal cord or esophagus were recorded. CONCLUSION: Overall survival, local control and freedom from local progression were comparable with the results reported with pure hyperfractionated radiotherapy. The overall rate of acute and late toxicity was acceptable. PMID- 15886272 TI - Changes in angiogenic growth factor levels after gefitinib treatment in non-small cell lung cancer. AB - BACKGROUND: To investigate the changes in angiogenic growth factor expression before and after gefitinib treatment, and the association between this expression and response to gefitinib treatment, we measured circulating levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), matrix metalloproteinase (MMP) -2 and -9, and tissue inhibitors of metalloproteinase (TIMP) -1 and -2 in patients with non-small cell lung cancer (NSCLC). METHODS: Serum and plasma samples were collected from 52 patients before and after gefitinib treatment. The levels of VEGF, bFGF, MMP-2, MMP-9, TIMP-1 and TIMP-2 were measured using a sandwich enzyme immunoassay kit. RESULTS: Of the 52 patients, 17 (32.7%) achieved a partial response, 19 (36.5%) had stable disease and 16 (30.8%) had progressive disease. The levels of VEGF, bFGF, MMP-2, MMP-9, TIMP-1 and TIMP-2 did not change significantly after gefitinib treatment, even in responders. The levels of VEGF in volunteers, responders and non-responders were 384 +/- 86.4, 404 +/- 94.3 and 719 +/- 99.8 pg/ml, respectively. The difference between volunteers and responders was not significant (P = 0.540), while the differences between volunteers and non-responders (P = 0.031), and responders and non-responders (P = 0.028) were significant. CONCLUSIONS: Although our results indicate that gefitinib treatment does not affect circulating levels of angiogenic growth factors even in patients who showed a response to gefitinib treatment, low levels of VEGF may predict response to gefitinib treatment in patients with NSCLC. PMID- 15886273 TI - Kallikrein gene transfer reduces renal fibrosis, hypertrophy, and proliferation in DOCA-salt hypertensive rats. AB - In DOCA-salt hypertension, renal kallikrein levels are increased and may play a protective role in renal injury. We investigated the effect of enhanced kallikrein levels on kidney remodeling of DOCA-salt hypertensive rats by systemic delivery of adenovirus containing human tissue kallikrein gene. Recombinant human kallikrein was detected in the urine and serum of rats after gene delivery. Kallikrein gene transfer significantly decreased DOCA- and salt-induced proteinuria, glomerular sclerosis, tubular dilatation, and luminal protein casts. Sirius red staining showed that kallikrein gene transfer reduced renal fibrosis, which was confirmed by decreased collagen I and fibronectin levels. Furthermore, kallikrein gene delivery diminished myofibroblast accumulation in the interstitium of the cortex and medulla, as well as transforming growth factor (TGF)-beta1 immunostaining in glomeruli. Western blot analysis and ELISA verified the decrease in immunoreactive TGF-beta1 levels. Kallikrein gene transfer also significantly reduced kidney weight, glomerular size, proliferating tubular epithelial cells, and macrophages/monocytes. Reduction of proliferation and hypertrophy was associated with reduced levels of the cyclin-dependent kinase inhibitor p27(Kip1), and the phosphorylation of c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). The protective effects of kallikrein were accompanied by increased urinary nitrate/nitrite and cGMP levels, and suppression of superoxide formation. These results indicate that kallikrein protects against mineralocorticoid-induced renal fibrosis glomerular hypertrophy, and renal cell proliferation via inhibition of oxidative stress, JNK/ERK activation, and p27(Kip1) and TGF-beta1 expression. PMID- 15886274 TI - Vasopressin increases urea permeability in the initial IMCD from diabetic rats. AB - In normal rats, vasopressin and hyperosmolality enhance urea permeability (P(urea)) in the terminal, but not in the initial inner medullary collecting duct (IMCD), a process thought to occur through the UT-A1 urea transporter. In the terminal IMCD, UT-A1 is detected as 97- and 117-kDa glycoproteins. However, in the initial IMCD, only the 97-kDa form is detected. During streptozotocin-induced diabetes mellitus, UT-A1 protein abundance is increased, and the 117-kDa UT-A1 glycoprotein appears in the initial IMCD. We hypothesize that the 117-kDa glycoprotein mediates the vasopressin- and osmolality-induced changes in P(urea). Thus, in the present study, we measured P(urea) in in vitro perfused initial IMCDs from diabetic rats by imposing a 5 mM bath-to-lumen urea gradient without any osmotic gradient. Basal P(urea) was similar in control vs. diabetic rats (3 +/- 1 vs. 5 +/- 1 x 10(-5) cm/s, n = 4, P = not significant). Vasopressin (10 nM) significantly increased P(urea) to 16 +/- 5 x 10(-5) cm/s (n = 4, P < 0.05) in diabetic but not in control rats. Forskolin (10 microM, adenylyl cyclase activator) also significantly increased P(urea) in diabetic rats. In contrast, increasing osmolality to 690 mosmol/kg H2O did not change P(urea) in diabetic rats. We conclude that initial IMCDs from diabetic rats have vasopressin- and forskolin-, but not hyperosmolality-stimulated P(urea). The appearance of vasopressin-stimulated P(urea) in initial IMCDs correlates with an increase in UT A1 protein abundance and the appearance of the 117-kDa UT-A1 glycoprotein in this region during diabetes. This suggests that the 117-kDa UT-A1 glycoprotein is necessary for vasopressin-stimulated urea transport. PMID- 15886275 TI - PPARgamma agonists exert antifibrotic effects in renal tubular cells exposed to high glucose. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) are ligand-activated transcription factors that regulate cell growth, inflammation, lipid metabolism, and insulin sensitivity. We recently demonstrated that PPARgamma agonists limit high glucose-induced inflammation in a model of proximal tubular cells (PTC; Panchapakesan U, Pollock CA, and Chen XM. Am J Physiol Renal Physiol 287: F528 F534, 2004). However, the role of PPARgamma in the excess extracellular matrix production is largely unknown. We evaluated the effect of 24- to 48-h 8 microM l 805645 or 10 microM pioglitazone on 25 mM D-glucose-induced markers of fibrosis in HK-2 cells. High D-glucose induced nuclear binding of activator protein-1 (AP 1) to 140.8 +/- 10.9% (P < 0.05), which was attenuated with L-805645 and pioglitazone to 82.3 +/- 14.4 (P < 0.01 vs. high D-glucose) and 99.3 +/- 12.2% (P < 0.05 vs. high D-glucose), respectively. High D-glucose increased total production of transforming growth factor (TGF)-beta(1) 139.6 +/- 6.5% (P < 0.05), which was reversed with L-805645 and pioglitazone to 68.73 +/- 5.7 (P < 0.01 vs. high D-glucose) and 112 +/- 13.6% (P < 0.05 vs. high D-glucose). L-805645 and pioglitazone reduced high d-glucose-induced fibronectin from 156.0 +/- 24.9 (P < 0.05) to 81.9 +/- 16.0 and 57.4 +/- 12.7%, respectively (both P < 0.01 vs. high D glucose). Collagen IV was not induced by high d-glucose. L-805645 and pioglitazone suppressed collagen IV to 68.0 +/- 14.5 (P < 0.05) and 46.5 +/- 11.6% (P < 0.01) vs. high D-glucose, respectively. High D-glucose increased the nuclear binding of NF-kappaB to 167 +/- 22.4% (P < 0.05), which was not modified with PPARgamma agonists. In conclusion, PPARgamma agonists exert antifibrotic effects in human PTC in high glucose by attenuating the increase in AP-1, TGF beta(1), and the downstream production of the extracellular matrix protein fibronectin. PMID- 15886276 TI - Enhanced sodium-dependent extrusion of magnesium in mutant cells established from a mouse renal tubular cell line. AB - To study the regulatory mechanisms of intracellular Mg(2+) concentration ([Mg(2+)](i)) in renal tubular cells as well as in other cell types, we established a mutant strain of mouse renal cortical tubular cells that can grow in culture media with very high extracellular Mg(2+) concentrations ([Mg(2+)](o) > 100 mM: 101Mg-tolerant cells). [Mg(2+)](i) was measured with a fluorescent indicator furaptra (mag-fura 2) in wild-type and 101Mg-tolerant cells. The average level of [Mg(2+)](i) in the 101Mg-tolerant cells was kept lower than that in the wild-type cells either at 51 mM or 1 mM [Mg(2+)](o). When [Mg(2+)](o) was lowered from 51 to 1 mM, the decrease in [Mg(2+)](i) was significantly faster in the 101Mg-tolerant cells than in the wild-type cells. These differences between the 101Mg-tolerant cells and the wild-type cells were abolished in the absence of extracellular Na(+) or in the presence of imipramine, a known inhibitor of Na(+)/Mg(2+) exchange. We conclude that Na(+)-dependent Mg(2+) transport activity is enhanced in the 101Mg-tolerant cells. The enhanced Mg(2+) extrusion may prevent [Mg(2+)](i) increase to higher levels and may be responsible for the Mg(2+) tolerance. PMID- 15886277 TI - Micropuncture gene delivery and intravital two-photon visualization of protein expression in rat kidney. AB - Understanding molecular mechanisms of pathophysiology and disease processes requires the development of new methods for studying proteins in animal tissues and organs. Here, we describe a method for adenoviral-mediated gene transfer into tubule or endothelial cells of the rat kidney. The left kidney of an anesthetized rat was exposed and the lumens of superficial proximal tubules or vascular welling points were microinfused, usually for 20 min. The microinfusion solution contained adenovirus with a cDNA construct of either 1) Xenopus laevis actin depolymerizing factor/cofilin [XAC; wt-green fluorescent protein (GFP)], 2) actin GFP, or 3) GFP. Sudan black-stained castor oil, injected into nearby tubules, allowed us to localize the microinfused structures for subsequent visualization. Two days later, the rat was anesthetized and the kidneys were fixed for tissue imaging or the left kidney was observed in vivo using two-photon microscopy. Expression of GFP and GFP-chimeric proteins was clearly seen in epithelial cells of the injected proximal tubules and the expressed proteins were localized similarly to their endogenously expressed counterparts. Only a minority of the cells in the virally exposed regions, however, expressed these proteins. Endothelial cells also expressed XAC-GFP after injection of the virus cDNA construct into vascular welling points. An advantage of the proximal tubule and vascular micropuncture approaches is that only minute amounts of virus are required to achieve protein expression in vivo. This micropuncture approach to gene transfer of the virus cDNA construct and intravital two-photon microscopy should be applicable to study of the behavior of any fluorescently tagged protein in the kidney and shows promise in studying renal physiology and pathophysiology. PMID- 15886278 TI - Knowledge-based computational search for genes associated with the metabolic syndrome. AB - MOTIVATION: A methodology to search for genes associated with multifactorial diseases by integrating the large amount of accumulated knowledge is seriously needed. A comprehensive understanding derived from a holistic view of gene relationship structures can be gained from our proposed analysis called the cross subspace analysis (CSA). In this analysis, gene objects are generated by machine learning using their term occurrence patterns in MEDLINE abstracts and the degree of relationship between gene objects is quantified by matching these patterns. RESULTS: Structuralization of relationships of a set of genes was performed using CSA, which were retrieved using the terms, 'obesity', 'diabetes', 'hypertriglyceridemia' and 'hypertension' that refer to diseases comprising metabolic syndrome, on a 2D plane inferring important biomedical concepts from the gene distribution. Then, we prioritized the significance of 6131 well annotated human genes in terms of the distance on the plane from the centroid of 'metabolic syndrome'-related genes distribution. The validity was confirmed by comparing the knowledge extracted by the ordering with existing medical knowledge. PMID- 15886279 TI - SCHIP: statistics for chromosome interphase positioning based on interchange data. AB - MOTIVATION: The position of chromosomes in the interphase nucleus is believed to be associated with a number of biological processes. Here, we present a web-based application that helps analyze the relative position of chromosomes during interphase in human cells, based on observed radiogenic chromosome aberrations. The inputs of the program are a table of yields of pairwise chromosome interchanges and a proposed chromosome geometric cluster. Each can either be uploaded or selected from provided datasets. The main outputs are P-values for the proposed chromosome clusters. SCHIP is designed to be used by a number of scientific communities interested in nuclear architecture, including cancer and cell biologists, radiation biologists and mathematical/computational biologists. PMID- 15886280 TI - Differential and trajectory methods for time course gene expression data. AB - MOTIVATION: The issue of high dimensionality in microarray data has been, and remains, a hot topic in statistical and computational analysis. Efficient gene filtering and differentiation approaches can reduce the dimensions of data, help to remove redundant genes and noises, and highlight the most relevant genes that are major players in the development of certain diseases or the effect of drug treatment. The purpose of this study is to investigate the efficiency of parametric (including Bayesian and non-Bayesian, linear and non-linear), non parametric and semi-parametric gene filtering methods through the application of time course microarray data from multiple sclerosis patients being treated with interferon-beta-1a. The analysis of variance with bootstrapping (parametric), class dispersion (semi-parametric) and Pareto (non-parametric) with permutation methods are presented and compared for filtering and finding differentially expressed genes. The Bayesian linear correlated model, the Bayesian non-linear model the and non-Bayesian mixed effects model with bootstrap were also developed to characterize the differential expression patterns. Furthermore, trajectory clustering approaches were developed in order to investigate the dynamic patterns and inter-dependency of drug treatment effects on gene expression. RESULTS: Results show that the presented methods performed significant differently but all were adequate in capturing a small number of the potentially relevant genes to the disease. The parametric method, such as the mixed model and two Bayesian approaches proved to be more conservative. This may because these methods are based on overall variation in expression across all time points. The semi parametric (class dispersion) and non-parametric (Pareto) methods were appropriate in capturing variation in expression from time point to time point, thereby making them more suitable for investigating significant monotonic changes and trajectories of changes in gene expressions in time course microarray data. Also, the non-linear Bayesian model proved to be less conservative than linear Bayesian correlated growth models to filter out the redundant genes, although the linear model showed better fit than non-linear model (smaller DIC). We also report the trajectories of significant genes-since we have been able to isolate trajectories of genes whose regulations appear to be inter-dependent. PMID- 15886281 TI - Evidence for an inflammatory process in age-related macular degeneration gains new support. PMID- 15886282 TI - Studies of yeast oligosaccharyl transferase subunits using the split-ubiquitin system: topological features and in vivo interactions. AB - Oligosaccharyl transferase (OT) catalyzes the cotranslational N-glycosylation of nascent polypeptides in the endoplasmic reticulum in all eukaryotic systems. Due to the inherent difficulty in characterizing this membrane protein complex, the mode of enzymatic action has not been resolved. Here, we used a membrane protein two-hybrid approach, the split-ubiquitin system, to address two aspects of the enzyme complex in yeast: the topological features, as well as the in vivo interactions of all of the components. We investigated the N- and C-terminal orientation of these proteins and the presence or the absence of a cleavable signal sequence at their N termini. We found that Ost2p and Stt3p have only their N terminus located in the cytosol, whereas Ost3p and Swp1p have only their C terminus oriented in the cytosol. In the case of Ost5p and Ost6p, both their N and C termini are present in the cytosol. These findings also suggested that Ost2p, Stt3p, Ost5p, and Ost6p do not have a cleavable N-terminal signal sequence. The pairwise analysis of in vivo interactions among all of the OT subunits demonstrated that OT subunits display specific interactions with each other in a functional complex. By comparing this interaction pattern with that detected in vitro in a nonfunctional complex, we proposed that a distinct conformation rearrangement takes place when the enzyme complex changes from the nonfunctional state to the activated functional state. This finding is consistent with earlier work by others indicating that OT exhibits allosteric properties. PMID- 15886283 TI - The success of animal invaders. PMID- 15886284 TI - Mice lacking multidrug resistance protein 3 show altered morphine pharmacokinetics and morphine-6-glucuronide antinociception. AB - Glucuronidation is a major detoxification pathway for endogenous and exogenous compounds in mammals that results in the intracellular formation of polar metabolites, requiring specialized transporters to cross biological membranes. By using morphine as a model aglycone, we demonstrate that multidrug resistance protein 3 (MRP3/ABCC3), a protein present in the basolateral membrane of polarized cells, transports morphine-3-glucuronide (M3G) and morphine-6 glucuronide in vitro. Mrp3(-/-) mice are unable to excrete M3G from the liver into the bloodstream, the major hepatic elimination route for this drug. This results in increased levels of M3G in liver and bile, a 50-fold reduction in the plasma levels of M3G, and in a major shift in the main disposition route for morphine and M3G, predominantly via the urine in WT mice but via the feces in Mrp3(-/-) mice. The pharamacokinetics of injected morphine-glucuronides are altered as well in the absence of Mrp3, and this results in a decreased antinociceptive potency of injected morphine-6-glucuronide. PMID- 15886285 TI - Predicting inhibitory drug-drug interactions and evaluating drug interaction reports using inhibition constants. AB - OBJECTIVE: To review the use of inhibitory constants (Ki) determined from in vitro experiments in the prediction of the significance of inhibitory drug-drug interactions (DDIs). DATA SOURCES: Searches of MEDLINE (1966-August 2004) and manual review of journals, conference proceedings, reference textbooks, and Web sites were performed using the key search terms cytochrome P450, drug-drug interaction, inhibition constant, and Ki. STUDY SELECTION AND DATA EXTRACTION: All articles identified from the data sources were evaluated, and information deemed relevant was included for this review. DATA SYNTHESIS: The cytochrome P450 isoenzymes factor prominently in the explanation of numerous DDIs. Although the regulation of these enzymes by one drug can affect the pharmacokinetics of other drugs, the consequences may not necessarily be significant either in terms of pharmacokinetic or clinical outcomes. Yet, many DDI monographs originate as unconfirmed case reports that implicate the influence of one drug on the CYP mediated metabolism of another, and these often uncorroborated mechanisms can eventually become regarded as dogma. One consequence of this process is the over prediction of potentially important DDIs. The pharmaceutical industry, Food and Drug Administration, and pharmaceutical scientists have developed a strategy for predicting the significance of inhibitory DDIs at the earliest possible stages of drug development based on a new chemical entity's Ki value, determined in vitro. CONCLUSIONS: We suggest that the use of Ki values of drugs purported to behave as CYP inhibitors be incorporated in the assessment of case reports that ascribe DDIs to inhibition of metabolism of one drug by another. PMID- 15886286 TI - Comparison of the effects of nitroglycerin and nitroprusside on transmitral Doppler flow parameters in patients with hypertensive urgency. AB - BACKGROUND: Sodium nitroprusside (NIP) and nitroglycerin (NIT) are frequently selected agents for acutely reducing blood pressure. However, it is not clear which agent is more efficacious in improving left ventricular filling pressure in hypertensive crises. OBJECTIVE: To compare the acute effects of nitroglycerin (NIT) and nitroprusside (NIP) on transmitral Doppler filling parameters in patients with hypertensive urgency. METHODS: We identified 37 patients from our emergency department with hypertensive urgency and left ventricular filling abnormalities. Hypertensive urgency was defined as a severe blood pressure elevation without evidence of progressive end-organ injury. Patients were randomized to receive an infusion of NIT or NIP. NIT was infused at a starting dose of 10 microg/min; NIP was infused at a starting dose of 0.25 microg/kg/min. The infusion rates were adjusted to decrease mean arterial pressure by 25%, and this reduction was obtained within 2 hours in all patients. Diastolic filling parameters were measured by using echocardiography before and after treatment. Pulsed-wave Doppler transmitral flow velocities were used. Early diastolic flow, atrial contraction signal, early diastolic flow/atrial contraction signal, deceleration time, and isovolumetric relaxation time (IVRT) were measured. RESULTS: There were no differences between groups in baseline demographic and echocardiographic parameters. Blood pressure decreased significantly in both treatment groups. In posttreatment echocardiographic examinations, atrial contraction signal, deceleration time, and IVRT were significantly decreased in both treatment groups. Early diastolic flow was significantly decreased in the NIT group. There were no significant differences between the groups in terms of posttreatment early diastolic flow, atrial contraction signal, deceleration time, and IVRT values. CONCLUSIONS: In hypertensive urgency with left ventricular filling abnormalities, reduction of blood pressure associated with NIT or NIP treatment may improve transmitral Doppler filling parameters. There were no differences demonstrated between the 2 agents. PMID- 15886287 TI - Achievements in public health: elimination of rubella and congenital rubella syndrome-US, 1969-2004. PMID- 15886288 TI - Influenza vaccine prebooking and distribution strategies for the 2005-2006 influenza season. PMID- 15886289 TI - Extensive prolongation of aPTT with argatroban in an elderly patient with improving renal function, normal hepatic enzymes, and metastatic lung cancer. AB - OBJECTIVE: To report a case of an elderly male with improving renal function and normal hepatic function who sustained an elevated activated partial thromboplastin time (aPTT) after an infusion of argatroban was discontinued. CASE SUMMARY: A 77-year-old white male with a history of heparin-induced thrombocytopenia (HIT) and metastatic lung disease was started on argatroban for treatment of a right upper-extremity deep vein thrombosis (DVT). The infusion was initiated at 2.0 microg/kg/min and was titrated to a goal aPTT of 60-80 seconds. Argatroban was discontinued due to an aPTT elevated to >100 seconds; the aPTT remained elevated for 130 hours after discontinuation of the infusion. DISCUSSION: Argatroban dose reductions in patients with impaired liver and renal function test values have been reported. Elderly subjects may have a prolonged clearance compared with young healthy subjects, although the duration of effect has not been established. As of April 18, 2005, the effect of liver metastasis on argatroban pharmacokinetics in the setting of normal liver function enzyme levels has not been reported. An objective causality assessment using the Naranjo probability scale showed that the prolonged aPTT was probably attributable to argatroban. CONCLUSIONS: Clinicians should exercise caution when initiating argatroban at a dose of 2.0 microg/kg/min in elderly patients with underlying comorbidities, such as metastatic disease and renal impairment, since this may lead to excessive and prolonged anticoagulation and increased risk of bleeding. PMID- 15886290 TI - Hypersensitivity reactions to oxaliplatin in two asian patients. AB - OBJECTIVE: To report 2 cases of hypersensitivity reactions associated with oxaliplatin treatment in Asian patients. CASE SUMMARIES: A 33-year-old Chinese woman received adjuvant oxaliplatin in combination with fluorouracil and leucovorin. Shortly during her sixth infusion, she developed a severe hypersensitivity reaction. Despite prophylactic measures, she developed another reaction of similar severity during her subsequent infusion and was not further rechallenged with oxaliplatin. The second case involved a 45-year-old Malay woman who received oxaliplatin, fluorouracil, and leucovorin for metastatic colorectal cancer. Shortly during her ninth infusion, she developed a mild hypersensitivity reaction. With prophylactic measures, she developed less marked reactions with her subsequent 3 infusions. DISCUSSION: Hypersensitivity reactions to oxaliplatin have been reported to be between 12% and 16% in the Western population. As of April 20, 2005, there are only 2 previously published reports of hypersensitivity reactions to oxaliplatin in 6 Asian patients. The incidence rates of such reactions in different Asian ethnic groups could vary. The cumulative dose, time of exposure to oxaliplatin, and clinical features were variable and unpredictable in all of the reported Asian patients who developed hypersensitivity reactions. An objective causality assessment using the Naranjo probability scale revealed that oxaliplatin was the highly probable cause of hypersensitivity in the 2 Asian patients reported by our center. CONCLUSIONS: Patients who develop mild to moderate reactions can be rechallenged with the drug administered as a slow infusion with prophylactic measures. Desensitization may allow patients who experience severe hypersensitivity reactions to oxaliplatin to further receive effective therapy for their colorectal cancer. PMID- 15886291 TI - The effect of creatine intake on renal function. AB - OBJECTIVE: To examine the effect of creatine supplementation on renal function and estimates of creatinine clearance. DATA SOURCES: A MEDLINE search was conducted (1966-September 2004) using the key terms creatine, creatinine, kidney function tests, drug toxicity, and exercise. Relevant articles were cross referenced to screen for additional information. DATA SYNTHESIS: Supplementation with creatine, an unregulated dietary substance, is increasingly common in young athletes. To date, few studies have evaluated the impact of creatine on renal function and estimates of creatinine clearance. Because creatine is converted to creatinine in the body, supplementation with large doses of creatine may falsely elevate creatinine concentrations. Five studies have reported measures of renal function after acute creatine ingestion and 4 after chronic ingestion. All of these studies were completed in young healthy populations. Following acute ingestion (4-5 days) of large amounts of creatine, creatinine concentrations increased slightly, but not to a clinically significant concentration. Creatinine is also only minimally affected by longer creatine supplementation (up to 5.6 y). CONCLUSIONS: Creatine supplementation minimally impacts creatinine concentrations and renal function in young healthy adults. Although creatinine concentrations may increase after long periods of creatine supplementation, the increase is extremely limited and unlikely to affect estimates of creatinine clearance and subsequent dosage adjustments. Further studies are required in the elderly and patients with renal insufficiency. PMID- 15886292 TI - Transporters and their impact on drug disposition. AB - OBJECTIVE: To review the recent advances in knowledge about human transporters and their effect on drug disposition. DATA SOURCES: A MEDLINE search (1996-March 2005) was performed to identify pertinent literature on human transporters and their impact on drug disposition. Additional articles were identified from a manual search of the references of retrieved articles. STUDY SELECTION AND DATA EXTRACTION: Based on the identified studies, data were extracted on the impact of transporters on drug absorption, distribution, and elimination. DATA SYNTHESIS: The pharmacokinetic disposition of drugs is known to be influenced by metabolic enzymes, kidney function, and transporters. Recent research on human transporters has greatly advanced our understanding of their diversity and importance in drug disposition. In particular, members of the multidrug resistance family of transporters (MDR, MRP) are present in organs and tissues throughout the body and are known to significantly affect the absorption, distribution, and elimination of commonly prescribed drugs. A growing number of studies now demonstrate that alterations in transporter function as a result of drug interactions or genetic polymorphisms may explain a significant portion of the variability in treatment response for certain drugs. CONCLUSIONS: Human transporters contribute significantly to the pharmacokinetic disposition of drugs. Knowledge of substrates, inducers, and inhibitors of these transporters is necessary to ensure optimal patient outcomes. PMID- 15886293 TI - Levofloxacin-induced fatal toxic epidermal necrolysis. PMID- 15886294 TI - Phenazopyridine-induced sulfhemoglobinemia. AB - OBJECTIVE: To report a case of sulfhemoglobinemia in a patient receiving phenazopyridine for a urinary tract infection. CASE SUMMARY: A 63-year-old white woman presented to the emergency department with complaints of fatigue and bluish discoloration of her body that had gradually progressed over the previous 6-8 weeks. About 4 months prior to presenting to the emergency department, she had started taking phenazopyridine, an over-the-counter medication for symptoms of dysuria. Because the cyanosis did not improve after the patient received oxygen and methylene blue, sulfhemoglobinemia was suspected and confirmed by spectrophotometer analysis. DISCUSSION: Sulfhemoglobin is a green-pigmented molecule containing a sulfur atom in one or more of the porphyrin rings. It is a rare cause of cyanosis, which is usually drug induced. Sulfhemoglobinemia is suspected when a cyanotic patient has normal to near-normal oxygen tension, laboratory reports of elevated methemoglobin, and does not respond to methylene blue therapy. Sulfhemoglobinemia is relatively rare, despite the widespread use of drugs that have been reported to cause it. Predisposing factors, such as chronic constipation, present in our patient, have been suggested as a source of hydrogen sulfide. CONCLUSIONS: This case of sulfhemoglobinemia, which occurred after the patient took phenazopyridine, is considered a probable adverse event according to the Naranjo probability scale. PMID- 15886295 TI - Low-dose warfarin for prevention of symptomatic thromboembolism after orthopedic surgery. AB - BACKGROUND: Warfarin dosing with a target international normalized ratio (INR) range of 1.5-2.5 has not been reported as adequate for venous thromboembolism (VTE) prophylaxis after total knee (TKR) and total hip replacement (THR) surgery. OBJECTIVE: To evaluate the rate of symptomatic VTE after TKR and THR surgery using a low-dose (INR 1.5-2.5) warfarin protocol started the evening before surgery compared with a literature cohort treated with enoxaparin. METHODS: TKR/THR patients treated with a 21-day low-dose warfarin protocol were followed via a consecutive observational design. Main outcome measures were symptomatic VTE and pulmonary embolism (PE), with major bleeds and death as secondary outcomes. Low-dose warfarin was compared with a literature cohort of patients treated with enoxaparin who received enoxaparin for a similar length of time and was evaluated for the same outcomes. Cohort event rates were derived as a weighted average using the DerSimonian model. RESULTS: VTE, PE, bleeds, and deaths in the low-dose warfarin group were 8 (1.04%), 4 (0.52%), 8 (1.04%), and 4 (0.52%), respectively. The cohort weighted average values were 35 (1.33%), 19 (0.72%), 65 (2.46%), and 18 (0.67%), respectively. Odds ratios for low-dose warfarin for VTE, PE, and VTE plus PE were 0.778 (95% CI 0.36 to 1.68), 0.717 (0.24 to 2.11), and 0.754 (0.41 to 1.42), respectively, all nonsignificant. Odds ratios for bleeds and death were 0.420 (0.20 to 0.87; p = 0.02) and 0.756 (0.26 to 2.24; NS), respectively. CONCLUSIONS: For this evaluation, low-dose warfarin was comparable to the enoxaparin cohort for development of VTE, PE, and VTE+PE. Incidences of bleeds in the enoxaparin cohort were significantly higher than in patients receiving low-dose warfarin. PMID- 15886296 TI - Repeated sequences in CASPASE-5 and FANCD2 but not NF1 are targets for mutation in microsatellite-unstable acute leukemia/myelodysplastic syndrome. AB - Microsatellite instability (MSI) in tumors is diagnostic for inactive DNA mismatch repair. It is widespread among some tumor types, such as colorectal or endometrial carcinoma, but is rarely found in leukemia. Therapy-related acute myeloid leukemia/myelodysplastic syndrome (tAML/MDS) is an exception, and MSI is frequent in tAML/MDS following cancer chemotherapy or organ transplantation. The development of MSI+ tumors is associated with an accumulation of insertion/deletion mutations in repetitive sequences. These events can cause inactivating frameshifts or loss of expression of key growth control proteins. We examined established MSI+ cell lines and tAML/MDS cases for frameshift-like mutations of repetitive sequences in several genes that have known, or suspected, relevance to leukemia. CASPASE-5, an acknowledged frameshift target in MSI+ gastrointestinal tract tumors, was frequently mutated in MSI+ cell lines (67%) and in tAML/MDS (29%). Frameshift-like mutations were also observed in the NF1 and FANCD2 genes that are associated with genetic conditions conferring a predisposition to leukemia. Both genes were frequent targets for mutation in MSI+ cell lines and colorectal carcinomas. FANCD2 mutations were also common in MSI+ tAML/MDS, although NF1 mutations were not observed. A novel FANCD2 polymorphism was also identified. PMID- 15886297 TI - Discovery of epigenetically masked tumor suppressor genes in endometrial cancer. AB - Realization that many tumor suppressor genes are silenced by epigenetic mechanisms has stimulated the discovery of novel tumor suppressor genes. We used a variety of research tools to search for genes that are epigenetically silenced in human endometrial cancers. Changes in global gene expression of the endometrial cancer cell line Ishikawa was analyzed after treatment with the demethylating agent 5-aza-2'-deoxycytidine combined with the histone deacetylase inhibitor suberoylanilide bishydroxamide. By screening over 22,000 genes, candidate tumor suppressor genes were identified. Additional microarray analysis and real-time reverse transcription-PCR of normal and cancerous endometrial samples and search for CpG islands further refined the list. Tazarotene-induced gene-1 (Tig1) and CCAAT/enhancer binding protein-alpha (C/ebpalpha) were chosen for further study. Expression of both genes was low in endometrial cancer cell lines and clinical samples but high in normal endometrial tissues. Bisulfite sequencing, restriction analysis, and/or methylation-specific PCR revealed aberrant methylation of the CpG island in the Tig1 gene of all 6 endometrial cancer cell lines examined and 4 of 18 clinical endometrial cancers, whereas the C/ebpalpha promoter remained unmethylated in endometrial cancers. Chromatin immunoprecipitation showed increased acetylated histone H3 bound to both Tig1 and C/ebpalpha genes after treatment with 5-aza-2'-deoxycytidine and/or suberoylanilide bishydroxamide. Forced expression of either TIG1 or C/EBPalpha led to significant growth reduction of Ishikawa cells. Our data suggest that C/ebpalpha and Tig1 function as tumor suppressor proteins in endometrial cancers and that their reexpression may be a therapeutic target. PMID- 15886298 TI - E1A specifically enhances sensitivity to topoisomerase IIalpha targeting anticancer drug by up-regulating the promoter activity. AB - DNA topoisomerases I and II (topo I and II) are nuclear enzymes involved in cellular replication and are targets for several anticancer drugs. We showed previously that E1A gene transfer enhanced the sensitivity of Ewing's sarcoma cells to the topo IIalpha targeting agents etoposide and Adriamycin in vitro and in vivo. To determine whether this effect was specific for topo IIalpha, we investigated the effect of E1A gene transfer on cell sensitivity to agents that target topo I and IIbeta. Transfecting TC71 human Ewing's sarcoma cells with an adenoviral vector containing the E1A gene enhanced their sensitivity to the topo IIalpha targeting agents etoposide (16-fold) and Adriamycin (8-fold). By contrast, E1A gene transfer did not affect cellular sensitivity to either amsacrine or camptothecin. Western blot analysis indicated that topo IIalpha protein levels increased 3.1-fold after E1A gene transfer, but topo I and IIbeta protein levels did not change. A plasmid containing topo IIalpha gene promoter with luciferase reporter gene was constructed to determine the effects of E1A gene transfer on the activity of the topo IIalpha promoter. E1A increased the activity of the topo IIalpha gene promoter by 3.5-fold relative to that of cells transfected with Ad-beta-gal. These results suggest that elevated topo IIalpha protein levels and enhanced sensitivity to topo IIalpha targeting agents were secondary to a direct effect of E1A on the topo IIalpha promoter. Combining E1A gene therapy with topo IIalpha targeting anticancer drugs may therefore have therapeutic benefit by increasing tumor cell sensitivity. PMID- 15886299 TI - Cisplatin sensitizes cancer cells to ionizing radiation via inhibition of nonhomologous end joining. AB - The combination of cisplatin and ionizing radiation (IR) treatment represents a common modality for treating a variety of cancers. These two agents provide considerable synergy during treatment, although the mechanism of this synergy remains largely undefined. We have investigated the mechanism of cisplatin sensitization to IR using a combination of in vitro and in vivo experiments. A clear synergistic interaction between cisplatin and IR is observed in cells proficient in nonhomologous end joining (NHEJ) catalyzed repair of DNA double strand breaks (DSB). In contrast, no interaction between cisplatin and IR is observed in NHEJ-deficient cells. Reconstituted in vitro NHEJ assays revealed that a site-specific cisplatin-DNA lesion near the terminus results in complete abrogation of NHEJ catalyzed repair of the DSB. These data show that the cisplatin-IR synergistic interaction requires the DNA-dependent protein kinase dependent NHEJ pathway for joining of DNA DSBs, and the presence of a cisplatin lesion on the DNA blocks this pathway. In the absence of a functional NHEJ pathway, although the cells are hypersensitive to IR, there is no synergistic interaction with cisplatin. PMID- 15886300 TI - Sphingosine-1-phosphate lyase regulates sensitivity of human cells to select chemotherapy drugs in a p38-dependent manner. AB - Resistance to cisplatin is a common problem that limits its usefulness in cancer therapy. Molecular genetic studies in the model organism Dictyostelium discoideum have established that modulation of sphingosine kinase or sphingosine-1-phosphate (S-1-P) lyase, by disruption or overexpression, results in altered cellular sensitivity to this widely used drug. Parallel changes in sensitivity were observed for the related compound carboplatin but not for other chemotherapy drugs tested. Sensitivity to cisplatin could also be potentiated pharmacologically with dimethylsphingosine, a sphingosine kinase inhibitor. We now have validated these studies in cultured human cell lines. HEK293 or A549 lung cancer cells expressing human S-1-P lyase (hSPL) show an increase in sensitivity to cisplatin and carboplatin as predicted from the earlier model studies. The hSPL-overexpressing cells were also more sensitive to doxorubicin but not to vincristine or chlorambucil. Studies using inhibitors to specific mitogen-activated protein kinases (MAPK) show that the increased cisplatin sensitivity in the hSPL-overexpressing cells is mediated by p38 and to a lesser extent by c-Jun NH2-terminal kinase MAPKs. p38 is not involved in vincristine or chlorambucil cytotoxicity. Measurements of MAPK phosphorylation and enzyme activity as well as small interfering RNA inhibition studies show that the response to the drug is accompanied by up-regulation of p38 and c-Jun NH2 terminal kinase and the lack of extracellular signal-regulated kinase up regulation. These studies confirm an earlier model proposing a mechanism for the drug specificity observed in the studies with D. discoideum and support the idea that the sphingosine kinases and S-1-P lyase are potential targets for improving the efficacy of cisplatin therapy for human tumors. PMID- 15886301 TI - Requirement of activated Cdc42-associated kinase for survival of v-Ras transformed mammalian cells. AB - Activated Cdc42-associated kinase (ACK) has been shown to be an important effector molecule for the small GTPase Cdc42. We have shown previously an essential role for Cdc42 in the transduction of Ras signals for the transformation of mammalian cells. In this report, we show that the ACK-1 isoform of ACK plays a critical role in transducing Ras-Cdc42 signals in the NIH 3T3 cells. Overexpression of a dominant-negative (K214R) mutant of ACK-1 inhibits Ras induced up-regulation of c-fos and inhibits the growth of v-Ras-transformed NIH 3T3 cells. Using small interfering RNA, we knocked down the expression of ACK-1 in both v-Ha-Ras-transformed and parental NIH 3T3 cells and found that down regulation of ACK-1 inhibited cell growth by inducing apoptosis only in v-Ha-Ras transformed but not parental NIH 3T3 cells. In addition, we studied the effect of several tyrosine kinase inhibitors and found that PD158780 inhibits the kinase activity of ACK-1 in vitro. We also found that PD158780 inhibits the growth of v Ha-Ras-transformed NIH 3T3 cells. Taken together, our results suggest that ACK-1 kinase plays an important role in the survival of v-Ha-Ras-transformed cells, suggesting that ACK-1 is a novel target for therapies directed at Ras-induced cancer. PMID- 15886302 TI - Eosinophil granulocytes are activated during the remission phase of ulcerative colitis. AB - AIM: The aim of this study was to establish a method of investigating intestinal eosinophil and neutrophil granulocytes by flow cytometry, and to compare the distribution and activity of these cells in different stages of ulcerative colitis (UC). METHODS: Biopsy samples were taken from six locations of the entire colon and from the terminal ileum in 10 patients with active total UC, 10 patients with inactive total UC, eight patients with active distal UC, and 11 control subjects. Cell suspensions from biopsies and from peripheral blood were incubated with fluorophore conjugated monoclonal antibodies. The use of scatter plot-gating and specific antibodies was established in a flow cytometry assay. RESULTS: Eosinophils were more numerous and more active in patients with active UC than in controls. Interestingly, during inactive UC, the number of activated eosinophils was even larger. Eosinophil activity was high in the rectum of patients with distal colitis but was also slightly elevated in the proximal colon. Neutrophils were increased in number and activity during active but not inactive UC. In patients with distal colitis, activated neutrophils were only found in the sigmoid colon and rectum. CONCLUSION: With this method, we confirm that neutrophils participate in the inflammatory process during active UC, and that they express a resting phenotype during remission. The finding of activated eosinophils in inflamed intestine strengthens the view of these cells as proinflammatory and tissue damaging. Nevertheless, our new finding of high eosinophil activation during inactive UC suggests that eosinophils play a role in repair of injured epithelium. PMID- 15886303 TI - A targeted approach to reducing lung cancer mortality. PMID- 15886304 TI - Epidemiology of lung cancer: looking to the future. AB - In the United States, the 20th century witnessed the emergence of a lung cancer epidemic that peaked and began to decline by the century's end, a decline that continues today. However, lung cancer continues to be an unabating pandemic. In research carried out over the last half of the 20th century, many factors were causally associated with lung cancer and studies were implemented to identify determinants of susceptibility to these factors. Cigarette smoking was identified as the single most predominant cause of the lung cancer epidemic, but other causes were found, including workplace agents (eg, asbestos, arsenic, chromium, nickel, and radon) and other environmental factors (passive smoking, indoor radon, and air pollution). Contemporary epidemiologic research on lung cancer now focuses on a new set of issues, primarily related to susceptibility to the well identified causal factors, particularly smoking, and on the consequences of changes in tobacco products for risks to smokers. Diet and the possibility of reducing risk through chemoprevention remain a focus of research emphasis through experimental and observational approaches. Questions have also been raised about possible differences in susceptibility to lung cancer by sex and race. Population patterns in smoking prevalence will continue to be the most powerful predictor of the future occurrence of lung cancer. Evaluation of recent US patterns in smoking prevalence indicates that for the next approximately 10 to 15 years, lung cancer rates will decrease, but will then level off starting in approximately 2030. Unless further reductions in the prevalence of cigarette smoking are achieved over the next decade, lung cancer will remain as an all too common, but avoidable, disease. PMID- 15886305 TI - Advances in the biology of lung cancer chemoprevention. AB - The heavy burden of lung cancer, which includes the highest worldwide mortality of any cancer, and its resistance to standard approaches (smoking cessation, screening, and therapy), have motivated an intense interest in chemoprevention of this disease. Randomized controlled trials of agents (including retinoids, beta carotene, and vitamin E) to prevent lung cancer have produced only disappointing clinical results to date. New, molecular-targeted approaches are advancing rapidly, however, with many promising targets and interactive signaling pathways for developing novel agents and combinatorial approaches in this setting. This promise is illustrated by recent studies of 15-hydroxyprostaglandin dehydrogenase, which plays a critical role in polyunsaturated fatty acid metabolism and (like another important target, prostacyclin) is downstream of cyclooxygenase-2. 15-hydroxyprostaglandin dehydrogenase degrades prostaglandin E(2), appears to have tumor suppressor activity, and can be induced both by peroxisome proliferator-activated receptor-gamma ligands and an epidermal growth factor receptor inhibitor. Other important targets/pathways include the insulin like growth factor axis, phosphoinositide 3-kinase pathway, cyclin D and E family members, and epigenetic events. Defining highest lung cancer risk (eg, establishing molecular risk models through long-term analyses of high-risk cohorts) will facilitate the clinical development of molecular-targeted prevention that will potentially reduce the enormous burden of lung cancer. PMID- 15886306 TI - New developments in lung cancer screening. AB - Lung cancer is the most lethal cancer in our society. Late diagnosis of this disease is a major problem and so recent favorable reports with spiral computed tomography screening of high-risk populations have rekindled interest in improving early lung cancer detections. The process of lung cancer screening is a complicated process that involves many component activities. Interest to date has heavily focused on the initial case identification, but more recent reports have suggested that the issues with case work-up and surgical management also bear closer consideration. Given the dynamic nature of spiral computed tomography scan development and the remarkable improvements in imaging resolution over the last decade, there is an urgent need for research to establish optimal clinical management of early lung cancer detected in a screening setting. PMID- 15886307 TI - Functional imaging in lung cancer. AB - Accurate detection of the presence and extent of disease is vital in the management of non-small-cell lung cancer. While computed tomography and magnetic resonance imaging tend to be the routine diagnostic modalities used in the management of lung cancer, there have been significant advances in the field of functional and molecular imaging. In this article, we review the performance of the functional imaging techniques that are currently available for the evaluation of non-small-cell lung cancer. The techniques range from evaluation of glucose metabolism in tumors with fluorodeoxyglucose, to evaluation of proliferation with fluorothymidine and evaluation of tumor hypoxia with agents such as fluoromisonidazole. Magnetic resonance imaging with an emphasis on dynamic contrast enhancement of tumors as well as detecting of malignant lymph nodes with targeted contrast agents is discussed. Emerging technologies such as lung imaging fluorescence endoscopy are considered. The role of functional imaging in planning, predicting response to, and evaluating effects of, various therapies is explored. PMID- 15886308 TI - Lung cancer in women. AB - Lung cancer is the leading cause of cancer death in the United States and is responsible for 20,000 more deaths yearly in US women than breast cancer. Cigarette smoking is the major cause of lung cancer, and unfortunately, approximately 22 million US women smoke. Mounting evidence suggests that there are significant differences in lung cancer between the sexes. There is a difference in the histologic distribution of lung cancer, with glandular differentiation being more common in women. Genetic variation may account for differences in susceptibility, and hormonal and biologic factors may play a role in carcinogenesis. Lung cancer patients have few therapeutic options. A more thorough understanding of the heterogeneity of lung cancer across populations may lead to innovations in treatment and prevention strategies. PMID- 15886309 TI - Genomic and proteomic profiling of lung cancers: lung cancer classification in the age of targeted therapy. AB - Both proteomic and genomic methods offer promise for the classification of human lung carcinomas. This review summarizes the range of proteomic methods in development for lung cancer classification, and describes a number of recent analyses of messenger RNA expression in lung cancer. Multiple independent studies of mRNA expression profiles in lung adenocarcinoma have proven highly reproducible. Analyses of the relationship between expression profiles and tumor development and differentiation, the presence or absence of specific pathogenic mutations, patient prognosis and survival after surgical treatment, and specific histopathology all appear to be promising. PMID- 15886310 TI - Epidermal growth factor receptor mutations in non-small-cell lung cancer: implications for treatment and tumor biology. AB - The epidermal growth factor receptor (EGFR) has emerged as an attractive therapeutic target for patients with non-small-cell lung cancer (NSCLC). However, despite its almost universal presence in NSCLC tumors, therapeutic inhibition of EGFR has resulted in significant tumor regressions in only 10% to 20% of patients. Several investigations over the last 12 months have uncovered somatic mutations in EGFR that underlie the sensitivity to EGFR inhibitors. NSCLC tumors and cell lines with EGFR mutations are exquisitely sensitive to the EGFR tyrosine kinase inhibitors (TKIs), erlotinib and gefitinib, and are biologically distinct from other forms of NSCLC. Somatic mutations in EGFR are found more frequently in patients with adenocarcinomas, nonsmokers, patients of Asian ethnicity, and in females. EGFR mutation detection is now becoming clinically available and is being incorporated into clinical treatment decisions and into the design of future clinical trials. Mutations in K-ras, a mediator of EGFR signaling, are mutually exclusive with EGFR mutations, and are associated with resistance to EGFR TKIs. In addition, secondary mutations, conferring resistance to EGFR TKIs, in patients with primary EGFR mutations once sensitive to EGFR TKIs, are beginning to be identified. The frequency of EGFR mutations, their impact on NSCLC biology, clinical treatment, and clinical trial design, as well as methods and limitations for mutation detection, will be reviewed. PMID- 15886311 TI - Epidermal growth factor receptor inhibitors in the treatment of non-small-cell lung cancer. AB - Remarkable developments in the systemic treatment of advanced non-small-cell lung cancer have taken place in the past few years. Targeted therapies have been largely employed in patients with far advanced disease, and some of them have demonstrated consistent activity in this setting. Epidermal growth factor receptor (EGFR) inhibitors cause dramatic response in approximately 10% of white patients who had received prior chemotherapy. Responses are higher in Asians. These findings are at least partly caused by the substantially higher incidence of EGFR mutations in Asians compared with whites. Studies of EGFR inhibitors in combination chemotherapy in front-line therapy of advanced non-small-cell lung cancer have, however, failed to improve survival, and better understanding of interactions between chemotherapeutic agents and EGFR inhibitors will be essential in the development of more effective strategies. PMID- 15886312 TI - Angiogenesis and lung cancer: prognostic and therapeutic implications. AB - Lung cancer is the most common cause of cancer death worldwide, with most patients dying with metastatic disease. The prognosis for the majority of patients remains poor. It is evident that advances in the treatment of this and other tumor types will require new approaches, and recent research has focused on molecular-targeted therapies. A key therapeutic strategy is inhibition of specific processes essential for tumor vascular development (a concept known to be beneficial in colorectal cancer) and a range of such antiangiogenic agents are currently in development. The most promising of these target the proangiogenic vascular endothelial growth factor (VEGF), either by preventing VEGF-receptor binding or inhibiting downstream receptor signaling. However, other more direct approaches against tumor vasculature are also in development. Since antiangiogenic agents often exert an indirect, cytostatic effect, many are being evaluated in combination with conventional chemotherapies in order to optimize the anticancer effects of both strategies. Additionally, the combination of several antiangiogenic agents is also being explored. This has become possible given the large number of agents currently available. As part of this evaluation process, the assessment of surrogate markers of target inhibition and treatment effect is ongoing in the hope of identifying reliable surrogate markers to aid the development of this new generation of anticancer agents. PMID- 15886313 TI - Multimodality therapy for stage III non-small-cell lung cancer. AB - The treatment of stage III non-small-cell lung cancer has evolved over the last two decades, with combined-modality therapy the current standard of care. As a result, intermediate and long-term survival has improved for patients in this common stage category, compared to the poor outcomes achieved with the historical standard of once-daily radiation therapy alone. This review summarizes two decades of clinical research regarding bimodality and trimodality approaches for the heterogenous stage subsets within the stage III designation, discusses the rationale and status of prophylactic brain irradiation, and concludes with perspectives on progress and future directions. Chemotherapy plus radiotherapy given concurrently is the optimal treatment for the group of patients with advanced stage III disease. The potential role of a surgical resection following chemotherapy (with or without radiation) in this setting is still controversial. The only subsets for which trimodality treatments are clearly preferred include T4N0-1 disease and superior sulcus tumors. The other major stage III subgroup has a minimal disease burden with low tumor volume and/or microscopic N2 disease, thus technically could undergo a surgical resection upfront. Induction chemotherapy before surgery may yield a survival advantage, although the phase III trials in this area are not conclusive. Given the marked survival benefit from adjuvant chemotherapy after surgery in even earlier stages of non-small-cell lung cancer, the proper sequence of surgery and chemotherapy (before v after surgery) remains an important unresolved question in this subgroup. Furthermore, how to incorporate radiation therapy, as well as whether it should be given at all in this subset of patients, are other important issues actively under study in ongoing trials. PMID- 15886314 TI - Adjuvant chemotherapy in completely resected non-small-cell lung cancer. AB - Surgery alone has long been the standard treatment for patients with operable non small-cell lung cancer (NSCLC). However, despite complete resection, 5-year survival rates have been disappointing, with about 50% of patients eventually suffering relapse and death from disease. Randomized trials conducted in the 1980s hinted at a survival benefit for postoperative cisplatin-based regimens, but they were underpowered. A meta-analysis published in 1995 found a nonsignificant 13% reduction in the risk of death associated with cisplatin-based chemotherapy, with an increase of survival of 5% at 5 years. This led to renewed interest in adjuvant chemotherapy in resected NSCLC. Thousands of patients have been included in a new generation of randomized trials in the last 10 years. Most of these recent studies have now been reported and several have demonstrated a clear survival advantage for patients treated with platin-based adjuvant therapy. These results also suggest a greater benefit with modern two-drug regimens. In view of the most recent data, postoperative platin-based chemotherapy can now be considered the standard of care for completely resected NSCLC patients with good performance status. PMID- 15886315 TI - Evolving concepts in the pathology and computed tomography imaging of lung adenocarcinoma and bronchioloalveolar carcinoma. AB - PURPOSE: To review recent advances in pathology and computed tomography (CT) of lung adenocarcinoma and bronchioloalveolar carcinoma (BAC). METHODS: A pathology/CT review panel of pathologists and radiologists met during a November 2004 International Association for the Study of Lung Cancer/American Society of Clinical Oncology consensus workshop in New York. The purpose was to determine if existing data was sufficient to propose modification of criteria for adenocarcinoma and BAC as newly published in the 2004 WHO Classification of Lung Tumors, and to address the pathologic/radiologic concept of diffuse/multicentric BAC. RESULTS: Solitary small, peripheral BACs have an excellent prognosis. Most lung adenocarcinomas with a BAC pattern are not pure BAC, but rather adenocarcinoma, mixed subtype with invasive patterns. This applies to tumors presenting with a diffuse/multinodular as well as solitary nodule pattern. The percent of BAC versus invasive components in lung adenocarcinomas appears to be prognostically important. However, a consensus definition of "minimally invasive" BAC with a favorable prognosis could not be achieved. While recognition of a BAC component is possible, the diagnosis of BAC with exclusion of invasive adenocarcinoma cannot be made by small biopsy or cytology specimens. CONCLUSION: There is a need to work toward a mutual understanding and consensus between pathologists, clinicians, and researchers with the use of the term BAC versus adenocarcinoma. Future studies should make some attempt to quantitate these components and/or other features such as size of scar, size of invasive component, or pattern of invasion. Hopefully, this work will allow definition of a category of adenocarcinoma, mixed subtype with predominant BAC/minimal invasion and a favorable prognosis. PMID- 15886316 TI - Systemic therapy of advanced bronchioloalveolar cell carcinoma: challenges and opportunities. AB - Bronchioloalveolar cell carcinoma (BAC) has fascinated physicians with its unique epidemiology, pathology, clinical manifestations, and natural history when compared with other non-small-cell lung cancer (NSCLC) subtypes. However, the relative rarity of pure BAC as defined by the WHO, and the inconsistent definitions used in various series, has limited systematic study of this entity. Retrospective and prospective studies suggest that patients with BAC treated with cytotoxic chemotherapy have a longer median survival than those with other subtypes of NSCLC. However, the widely accepted view that BAC is less chemosensitive than other NSCLCs is not clearly supported by the small body of available literature. Antitumor activity of cytotoxic agents and the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors has been documented in phase II trials but no phase III trials have been conducted in this disease. The observation that profound responses to gefitinib and erlotinib often occurred in NSCLC patients with BAC, and that EGFR tyrosine kinase domain mutations were identified in large part by careful study of such patients, serves as a paradigm for translational research in this disease. The recognition that pure BAC and adenocarcinoma with BAC features behave similarly and as such represent a relatively common entity will facilitate accrual to BAC specific studies. Alternatively, stratification of these histologic subtypes in broader clinical trials in NSCLC is warranted. However, for either strategy to succeed in advancing our understanding of the molecular biology of BAC, it must be accompanied by central pathologic review with detailed classification of such, and of adequate tissue for measurement of known or putative targets of action of the agent under study. PMID- 15886317 TI - Comparison of genome profiles for identification of distinct subgroups of diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphoma (DLBCL) comprises molecularly distinct subgroups such as activated B-cell-like (ABC) and germinal center B-cell-like (GCB) DLBCLs. We previously reported that CD5(+) and CD5(-)CD10(+) DLBCL constitute clinically relevant subgroups. To determine whether these 2 subgroups are related to ABC and GCB DLBCLs, we analyzed the genomic imbalance of 99 cases (36 CD5(+), 19 CD5( )CD10(+), and 44 CD5(-)CD10(-)) using array-based comparative genomic hybridization (CGH). Forty-six of these cases (22 CD5(+), 7 CD5(-)CD10(+), and 17 CD5(-)CD10(-)) were subsequently subjected to gene-expression profiling, resulting in their division into 28 ABC (19 CD5(+) and 9 CD5(-)CD10(-)) and 18 GCB (3 CD5(+), 7 CD5(-)CD10(+), and 8 CD5(-)CD10(-)) types. A comparison of genome profiles of distinct subgroups of DLBCL demonstrated that (1) ABC DLBCL is characterized by gain of 3q, 18q, and 19q and loss of 6q and 9p21, and GCB DLBCL is characterized by gain of 1q, 2p, 7q, and 12q; (2) the genomic imbalances characteristic of the CD5(+) and CD5(-)CD10(+) groups were similar to those of the ABC and GCB types, respectively. These findings suggest that CD5(+) and CD5( )CD10(+) subgroups are included, respectively, in the ABC and GCB types. Finally, when searching for genomic imbalances that affect patients' prognosis, we found that 9p21 loss (p16(INK4a) locus) marks the most aggressive type of DLBCL. PMID- 15886318 TI - A SCID-hu in vivo model of human Waldenstrom macroglobulinemia. AB - The preclinical evaluation of investigational agents for Waldenstrom macroglobulinemia (WM) has been limited by the lack of in vivo models that enable the use of explanted patient cells. We describe here a novel in vivo model of human WM in severe combined immunodeficient (SCID) mice implanted with human fetal bone chips (SCID-hu mice) into which WM cells from patient bone marrow are engrafted directly into the human bone marrow (huBM) microenvironment. WM cells in SCID-hu mice produced human monoclonal paraprotein (immunoglobulin M [IgM] and/or kappa or lambda chain) detectable in mice sera. Immunohistochemical analysis of human bone retrieved from SCID-hu mice showed infiltration with CD20+, IgM+, and monotypic light chain+ lymphoplasmacytic cells. Mast cells were observed to be associated with the infiltrate in these sections. Treatment of SCID-hu mice bearing WM with rituximab induced tumor regression, associated with a decrease in serum paraprotein. This model, therefore, recapitulates the in vivo biology of WM and allows the study of novel investigational drugs targeting WM cells in the huBM milieu. PMID- 15886319 TI - Time-course analysis of hepcidin, serum iron, and plasma cytokine levels in humans injected with LPS. AB - Hepatic peptide hormone hepcidin is the key regulator of iron metabolism and the mediator of anemia of inflammation. Previous studies indicated that interleukin-6 (IL-6) mediates hepcidin increase and consequent hypoferremia during inflammation. Here we used an in vivo human endotoxemia model to analyze the effects of lipopolysaccharide (LPS) as a more upstream inflammation activator. The temporal associations between plasma cytokines, hepcidin levels, and serum iron parameters were studied in 10 healthy individuals after LPS injection. IL-6 was dramatically induced within 3 hours after injection, and urinary hepcidin peaked within 6 hours, followed by a significant decrease in serum iron. Serum prohepcidin showed no significant change within a 22-hour time frame. These in vivo human results confirm the importance of the IL-6-hepcidin axis in the development of hypoferremia in inflammation and highlight the rapid responsiveness of this iron regulatory system. PMID- 15886320 TI - Altered NKG2D function in NK cells induced by chronic exposure to NKG2D ligand expressing tumor cells. AB - NKG2D is an activation receptor that allows natural killer (NK) cells to detect diseased host cells. The engagement of NKG2D with corresponding ligand results in surface modulation of the receptor and reduced function upon subsequent receptor engagement. However, it is not clear whether in addition to modulation the NKG2D receptor complex and/or its signaling capacity is preserved. We show here that the prolonged encounter with tumor cell-bound, but not soluble, ligand can completely uncouple the NKG2D receptor from the intracellular mobilization of calcium and the exertion of cell-mediated cytolysis. However, cytolytic effector function is intact since NKG2D ligand-exposed NK cells can be activated via the Ly49D receptor. While NKG2D-dependent cytotoxicity is impaired, prolonged ligand exposure results in constitutive interferon gamma (IFNgamma) production, suggesting sustained signaling. The functional changes are associated with a reduced presence of the relevant signal transducing adaptors DNAX-activating protein of 10 kDa (DAP-10) and killer cell activating receptor-associated protein/DNAX-activating protein of 12 kDa (KARAP/DAP-12). That is likely the consequence of constitutive NKG2D engagement and signaling, since NKG2D function and adaptor expression is restored to normal when the stimulating tumor cells are removed. Thus, the chronic exposure to tumor cells expressing NKG2D ligand alters NKG2D signaling and may facilitate the evasion of tumor cells from NK cell reactions. PMID- 15886321 TI - Bombay phenotype is associated with reduced plasma-VWF levels and an increased susceptibility to ADAMTS13 proteolysis. AB - ABO blood group is an important determinant of plasma von Willebrand factor antigen (VWF:Ag) levels, with lower levels in group O. Previous reports have suggested that ABO(H) sugars affect the susceptibility of VWF to ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type-1 repeats-13) cleavage. To further test this hypothesis, we collected plasma from individuals with the rare Bombay blood group. VWF:Ag levels were significantly lower in Bombay patients (median, 0.69 IU/mL) than in groups AB, A, or B (P < .05) and lower than in group O individuals (median, 0.82 IU/mL). Susceptibility of purified VWF fractions to recombinant ADAMTS13 cleavage, assessed using VWF collagen-binding assay (VWF:CB), was increased in Bombays compared with either group O or AB. Increasing urea concentration (0.5 to 2 M) increased the cleavage rate for each blood group but eliminated the differences between groups. We conclude that reduction in the number of terminal sugars on N-linked glycan increases susceptibility of globular VWF to ADAMTS13 proteolysis and is associated with reduced plasma VWF:Ag and VWF:CB levels. PMID- 15886322 TI - Identification and functional characterization of 2 variant alleles of the telomerase RNA template gene (TERC) in a patient with dyskeratosis congenita. AB - Heterozygous mutations of the human telomerase RNA template gene (TERC) have been described in patients with acquired aplastic anemia and the autosomal dominant form of dyskeratosis congenita (DKC). Patients with mutations in both TERC alleles have not yet been reported. Here, we report a patient with DKC who inherited 2 distinct TERC sequence variants from her parents; a deletion (216_229del) in one and a point mutation (37A>G) in the other allele of the TERC gene. Her marrow was hypocellular and showed an abnormal clone [46, XX t(7;21)(q34;q22)]. The telomere lengths in leukocytes of the patient and her relatives were shorter than those of the age-matched controls and were progressively shorter in subsequent generations of family members with the 216_229del allele. Telomerase enzymatic levels in lymphocytes from the patient were approximately half of those measured in healthy controls. The 216_229del mutation failed to reconstitute telomerase activity in transfected cells, but, when coexpressed with the 37A>G variant, telomerase activity was only modestly suppressed. These clinical and laboratory findings support the concept that telomerase levels in human hematopoietic stem cells are tightly controlled as even moderately reduced levels result in accelerated telomere shortening and eventual marrow failure. PMID- 15886323 TI - Wnts induce migration and invasion of myeloma plasma cells. AB - Multiple myeloma is an incurable form of lymphoid cancer characterized by accumulation of neoplastic plasma cells in the bone marrow cavity. Little is known about the mechanisms regulating myeloma cell movement within the bone marrow and metastasis to secondary sites. Herein, we identify multiple members of the wingless/int (Wnt) family as promoters of myeloma cell migration/invasion. Wnt-mediated migration was associated with the Wnt/RhoA pathway and did not necessitate signaling through beta-catenin. Activation of both RhoA and members of the protein kinase C (PKC) family, including PKCalpha, PKCbeta, and PKCmu, were required for induction of migration. Activated RhoA and PKCalpha, PKCbeta, and PKCmu appear to assemble in macromolecular signaling complexes that are associated with the cell membrane. These results suggest that Wnt responsiveness of myeloma plasma cells may be a significant factor in disease progression. PMID- 15886324 TI - Lentiviral shRNA silencing of murine bone marrow cell CCR2 leads to persistent knockdown of CCR2 function in vivo. AB - A major barrier in hematopoietic gene function studies is posed by the laborious and time-consuming generation of knockout mice with an appropriate genetic background. Here we present a novel lentivirus-based strategy for the in situ generation of hematopoietic knockdowns. A short hairpin RNA (shRNA) was designed targeting murine CC-chemokine receptor 2 (CCR2), which was able to specifically blunt CCR2 expression at the mRNA, protein, and functional levels in vitro. Reconstitution of irradiated recipient mice with autologous bone marrow that had been ex vivo transduced with shRNA lentivirus led to persistent down-regulation of CCR2 expression, which translated into a 70% reduction in CCR2-dependent recruitment of macrophages to an inflamed peritoneal cavity without noticeable side effects on related chemokine receptors or general inflammation status. These findings clearly demonstrate the potential of shRNA lentivirus-infected bone marrow transplantation as a rapid and effective method to generate hematopoietic knockdowns for leukocyte gene function studies. PMID- 15886325 TI - A rapamycin derivative (everolimus) controls proliferation through down regulation of truncated CCAAT enhancer binding protein {beta} and NF-{kappa}B activity in Hodgkin and anaplastic large cell lymphomas. AB - The immunosuppressive macrolide rapamycin and its derivative everolimus (SDZ RAD, RAD) inhibit the mammalian target of rapamycin (mTOR) signaling pathway. In this study, we provide evidence that RAD has profound antiproliferative activity in vitro and in NOD/SCID mice in vivo against Hodgkin lymphoma (HL) and anaplastic large cell lymphoma (ALCL) cells. Moreover, we identified 2 molecular mechanisms that showed how RAD exerts antiproliferative effects in HL and ALCL cells. RAD down-regulated the truncated isoform of the transcription factor CCAAT enhancer binding protein beta (C/EBPbeta), which is known to disrupt terminal differentiation and induce a transformed phenotype. Furthermore, RAD inhibited constitutive nuclear factor kappaB (NF-kappaB) activity, which is a critical survival factor of HL cells. Pharmacologic inhibition of the mTOR pathway by RAD therefore interferes with essential proliferation and survival pathways in HL and ALCL cells and might serve as a novel treatment option. PMID- 15886326 TI - GPVI and alpha2beta1 play independent critical roles during platelet adhesion and aggregate formation to collagen under flow. AB - The roles of the 2 major platelet-collagen receptors, glycoprotein VI (GPVI) and integrin alpha2beta1, have been intensely investigated using a variety of methods over the past decade. In the present study, we have used pharmacologic and genetic approaches to study human and mouse platelet adhesion to collagen under flow conditions. Our studies demonstrate that both GPVI and integrin alpha2beta1 play significant roles for platelet adhesion to collagen under flow and that the loss of both receptors completely ablates this response. Intracellular signaling mediated by the cytoplasmic adaptor Src homology 2 domain-containing leukocyte protein of 76 kDa (SLP-76) but not by the transmembrane adaptor linker for activation of T cells (LAT) is critical for platelet adhesion to collagen under flow. In addition, reduced GPVI receptor density results in severe defects in platelet adhesion to collagen under flow. Defective adhesion to collagen under flow is associated with prolonged tail-bleeding times in mice lacking one or both collagen receptors. These studies establish platelet-collagen responses under physiologic flow as the consequence of a close partnership between 2 structurally distinct receptors and suggest that both receptors play significant hemostatic roles in vivo. PMID- 15886327 TI - Persistent expression of factor VIII in vivo following nonprimate lentiviral gene transfer. AB - Hemophilia A is a clinically important coagulation disorder caused by the lack or abnormality of plasma coagulation factor VIII (FVIII). Gene transfer of the FVIII cDNA to hepatocytes using lentiviral vectors is a potential therapeutic approach. We investigated the efficacy of feline immunodeficiency virus (FIV)-based vectors in targeting hepatocytes and correcting FVIII deficiency in a hemophilia A mouse model. Several viral envelope glycoproteins were screened for efficient FIV vector pseudotyping and hepatocyte transduction. The GP64 glycoprotein from baculovirus Autographa californica multinuclear polyhedrosis virus pseudo-typed FIV efficiently and showed excellent hepatocyte tropism. The GP64-pseudotyped vector was stable in the presence of human or mouse complement. Inclusion of a hybrid liver-specific promoter (murine albumin enhancer/human alpha1-antitrypsin promoter) further enhanced transgene expression in hepatocytes. We generated a GP64-pseudotyped FIV vector encoding the B domain-deleted human FVIII coding region driven by the liver-specific promoter, with 2 beneficial point mutations in the A1 domain. Intravenous vector administration conferred sustained FVIII expression in hemophilia A mice for several months without the generation of anti human FVIII antibodies and resulted in partial phenotypic correction. These findings demonstrate the utility of GP64-pseudotyped FIV lentiviral vectors for targeting hepatocytes to correct disorders associated with deficiencies of secreted proteins. PMID- 15886328 TI - Safety and efficacy of gemtuzumab ozogamicin in pediatric patients with advanced CD33+ acute myeloid leukemia. AB - This open-label, dose-escalation study evaluated the safety and efficacy of single-agent gemtuzumab ozogamicin, a humanized anti-CD33 antibody-targeted chemotherapeutic agent, for pediatric patients with multiple relapsed or primary refractory acute myeloid leukemia (AML). Twenty-nine children 1 to 16 years of age (relapsed disease, 19; refractory disease, 10) received gemtuzumab ozogamicin ranging from 6 to 9 mg/m2 per dose for 2 doses (separated by 2 weeks) infused over 2 hours. All patients had anticipated myelosuppression. Other toxicities included grade 3/4 hyperbilirubinemia (7%) and elevated hepatic transaminase levels (21%); the incidence of grade 3/4 mucositis (3%) or sepsis (24%) was relatively low. One patient treated at 9 mg/m2 developed veno-occlusive disease (VOD) of the liver and defined the dose-limiting toxicity. Thirteen patients underwent hematopoietic stem-cell transplantation less than 3.5 months after the last dose of gemtuzumab ozogamicin; 6 (40%) developed VOD. Eight of 29 (28%) patients achieved overall remission. Remissions were comparable in patients with refractory (30%) and relapsed (26%) disease. Mean multidrug resistance-protein mediated drug efflux was significantly lower in the leukemic blasts of patients achieving remission (P < .005). Gemtuzumab ozogamicin was relatively well tolerated at 6 mg/m2 for 2 doses and was equally effective in patients with refractory and relapsed disease. Further studies in combination with standard induction therapy for childhood AML are warranted. PMID- 15886329 TI - Neutrophils rapidly migrate via lymphatics after Mycobacterium bovis BCG intradermal vaccination and shuttle live bacilli to the draining lymph nodes. AB - The early innate response after Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccination is poorly characterized but probably decisive for subsequent protective immunity against tuberculosis. Therefore, we vaccinated mice with fluorescent BCG strains in the ear dorsum, as a surrogate of intradermal vaccination in humans. During the first 3 days, we tracked BCG host cells migrating out of the dermis to the auricular draining lymph nodes (ADLNs). Resident skin dendritic cells (DCs) or macrophages did not play a predominant role in early BCG capture and transport to ADLNs. The main BCG host cells rapidly recruited both in the dermis and ADLNs were neutrophils. Fluorescent green or red BCG strains injected into nonoverlapping sites were essentially sheltered by distinct neutrophils in the ADLN capsule, indicating that neutrophils had captured bacilli in peripheral tissue and transported them to the lymphoid organ. Strikingly, we observed BCG-infected neutrophils in the lumen of lymphatic vessels by confocal microscopy on ear dermis. Fluorescence-labeled neutrophils injected into the ears accumulated exclusively into the ipsilateral ADLN capsule after BCG vaccination. Thus, we provide in vivo evidence that neutrophils, like DCs or inflammatory monocytes, migrate via afferent lymphatics to lymphoid tissue and can shuttle live microorganisms. PMID- 15886330 TI - Muscles of mice deficient in alpha-sarcoglycan maintain large masses and near control force values throughout the life span. AB - alpha-Sarcoglycan-deficient (Sgca-null) mice provide potential for elucidating the pathogenesis of limb girdle muscular dystrophy type 2D (LGMD 2D) as well as for studying the effectiveness of therapeutic strategies. Skeletal muscles of Sgca-null mice demonstrate an early onset of extensive fiber necrosis, degeneration, and regeneration, but the progression of the pathology and the effects on muscle structure and function throughout the life span are not known. Thus the phenotypic accuracy of the Sgca-null mouse as a model of LGMD 2D has not been fully established. To investigate skeletal muscle structure and function in the absence of alpha-sarcoglycan throughout the life span, we analyzed extensor digitorum longus and soleus muscles of male and female Sgca-null and wild-type mice at 3, 6, 12, and 18 mo of age. Maximum isometric forces and powers were measured in vitro at 25 degrees C. Also determined were individual myofiber cross sectional areas and numbers, water content, and the proportion of the cross section occupied by connective tissue. Muscle masses were 40-100% larger for Sgca null compared with age- and gender-matched wild-type mice, with the majority of the increased muscle mass for Sgca-null mice attributable to greater connective tissue and water contents. Although the greater mass of muscles in Sgca-null mice was primarily noncontractile material, absolute forces and powers were maintained near control levels at all ages, indicating a successful adaptation to the deficiency in alpha-sarcoglycan not observed at any age in LGMD 2D patients. PMID- 15886331 TI - Uncoupling protein 1 in fish uncovers an ancient evolutionary history of mammalian nonshivering thermogenesis. AB - Uncoupling proteins (UCPs) increase proton leakage across the inner mitochondrial membrane. Thereby, UCP1 in brown adipose tissue dissipates proton motive force as heat. This mechanism of nonshivering thermogenesis is considered as a monophyletic trait of endothermic placental mammals that emerged about 140 million years ago and provided a crucial advantage for life in the cold. The paralogues UCP2 and UCP3 are probably not thermogenic proteins but convey mild uncoupling, which may serve to reduce the rate of mitochondrial reactive oxygen species production. Both are present in endotherms (mammals and birds), but so far only UCP2 has been identified in ectothermic vertebrates (fish and amphibia). The evolution of UCPs is of general interest in the search for the origin of mammalian UCP1-mediated nonshivering thermogenesis. We here show the presence of UCP1 and UCP3 in ectothermic teleost fish species using comparative genomics, phylogenetic inference, and gene expression analysis. In the common carp (Cyprinus carpio), UCP1 is predominantly expressed in the liver and strongly diminished in response to cold exposure, thus contrasting the cold-induced expression of mammalian UCP1 in brown adipose tissue. UCP3 mRNA is only found in carp skeletal muscle with expression levels increased fivefold in response to fasting. Our findings disprove the monophyletic nature of UCP1 in placental mammals and demonstrate that all three members of the core UCP family were already present before the divergence of ray-finned and lobe-finned vertebrate lineages about 420 million years ago. PMID- 15886332 TI - Gene expression profiling and phenotype analyses of S. cerevisiae in response to changing copper reveals six genes with new roles in copper and iron metabolism. AB - Exhaustive microarray time course analyses of Saccharomyces cerevisiae during copper starvation and copper excess reveal new aspects of metal-induced gene regulation. Aside from identifying targets of established copper- and iron responsive transcription factors, we find that genes encoding mitochondrial proteins are downregulated and that copper-independent iron transport genes are preferentially upregulated, both during prolonged copper deprivation. The experiments also suggest the presence of a small regulatory iron pool that links copper and iron responses. One hundred twenty-eight genes with putative roles in metal metabolism were further investigated by several systematic phenotype screens. Of the novel phenotypes uncovered, hsp12-Delta and arn1-Delta display increased sensitivity to copper, cyc1-Delta and crr1-Delta show resistance to high copper, vma13-Delta exhibits increased sensitivity to iron deprivation, and pep12-Delta results in reduced growth in high copper and low iron. Besides revealing new components of eukaryotic metal trafficking pathways, the results underscore the previously determined intimate links between iron and copper metabolism and mitochondrial and vacuolar function in metal trafficking. The analyses further suggest that copper starvation can specifically lead to downregulation of respiratory function to preserve iron and copper for other cellular processes. PMID- 15886333 TI - Genomic organization, expression, and function of bitter taste receptors (T2R) in mouse and rat. AB - Mammalian type 2 taste receptors (T2R) are a family of G protein-coupled receptors that mediate bitter signals in taste cells. In the present study, we compared the genomic organization of rodent T2R genes based on the recently completed mouse and rat genomes and examined tissue- and cell-specific expression of T2Rs. Both mouse and rat T2R families consist of 36 intact genes and at least 7 pseudogenes that are mapped to mouse chromosomes 15, 2, and 6 and to rat chromosomes 2, 3, and 4, respectively. All but two T2R genes are clustered on mouse chromosome 6 and rat chromosome 4 with virtually identical genomic organization. The orthologs of the first human T2R gene identified, mT2R119 and rT2R1, are located on mouse chromosome 15 and rat chromosome 2, whereas the novel rodent-specific T2R genes, mT2R134 and rT2R34, are located on mouse chromosome 2 and rat chromosome 3, respectively. Our results, using RT-PCR, demonstrate the presence of transcripts corresponding to the putative denatonium benzoate (DB) and phenylthiocarbamide (PTC) receptors in the antrum, fundus, and duodenum as well as in STC-1 and AR42J cells. The novel rodent-specific T2R gene (mT2R134 and rT2R34) was also expressed in these tissues and cell lines. The addition of DB, PTC, or cycloheximide to AR42J cells induced a rapid increase in the intracellular Ca(2+) concentration. The specificity of these effects is shown by the fact that these bitter stimuli did not induce any detectable Ca(2+) signaling in many other rodent or human cells that do not express receptors or G proteins implicated in bitter taste signaling. These results demonstrate that mouse and rat T2R genes are highly conserved in terms of genomic organization and tissue expression, suggesting that rodent T2Rs are evolved under similar dietary pressure and share bitter sensing functions in the lingual and gastrointestinal systems. PMID- 15886334 TI - Perchlorate in human milk: separating the science from sensationalism. PMID- 15886336 TI - Cycle of seeking, emotional outburst, and rest in newborns put to the breast. PMID- 15886338 TI - Breast fat and fallacies: more than 100 years of anatomical fantasy. AB - The authors studied the anatomy of 136 patients who underwent breast reduction surgery from 1998 to 2003 to determine the relationship of breast fat to the glandular tissue of the breast. Histological sections of freshly preserved breast tissue taken from representative patients were examined and compared to depictions of normal breast anatomy as portrayed in standard anatomical texts from the classic work of Sir Astley Cooper in 1845 to current publications such as Auberbach and Riordan's Breastfeeding and Human Lactation. Most texts portray little intermix of fat with the glandular tissue of the breast. Our studies confirm the texts that demonstrate the fat and glandular tissue to be inseparable and present in continuity with each other except in the subcutaneous plane where only fat is present. The implications of this anatomical fact as it relates to breast surgery and human lactation are discussed. PMID- 15886339 TI - Screening of virulence determinants in Enterococcus faecium strains isolated from breast milk. AB - In a previous study, the authors isolated lactic acid bacteria from breast milk of healthy mothers. Since some of the identified isolates belonged to the species Enterococcus faecium, the objective of this work was to evaluate their safety. The enterococcal strains were screened by polymerase chain reaction (PCR) and Southern hybridization for the presence of virulence determinants. The potential of the strains to acquire plasmids by conjugation was investigated by screening for genes involved in conjugation processes. Parallel, phenotypic assays were performed. Presence of genes conferring resistance to vancomycin was assessed by PCR. PCR amplifications and Southern hybridizations revealed that all the strains were clear of the majority of potential virulence determinants. None of the strains showed gelatinase activity, hemolysin production, or aggregation phenotype, and none carried the vanA or vanB genes. These findings suggest that milk of healthy mothers may be a source of avirulent E faecium isolates to the newborns. PMID- 15886340 TI - Primary and secondary mediators' influence on milk output in lactating mothers of preterm and term infants. AB - This study examined potential primary mediators, such as intended length to breastfeed, maternal education, income, and infant gestation, and secondary mediators, such as early frequency of breast stimulation, early milk output, and supplementation with artificial milks that may influence milk output in mothers of preterm and term infants the first 6 weeks postpartum. Analysis suggested that for mothers of a preterm infant (n = 95), the primary mediators, income and infant gestation, and the secondary mediators, early milk output/d and early frequency/d, accounted for 53.5% of the variance in milk output/d at week 6. For mothers of a term infant (n = 98), the primary mediator, income, and secondary mediators, early milk output/d and supplementation, accounted for 48.4% of the variance in milk output/d at week 6. Further research is needed to determine what early interventions may improve milk output in mothers at risk for lactation failure. PMID- 15886341 TI - The cup-versus-bottle debate: a theme from an ethnographic study of the supplementation of breastfed infants in hospital in the United kingdom. AB - This article reports 1 theme from an ethnographic study that aimed to describe the experiences, expectations, and beliefs of mothers and health care professionals concerning supplementation in a UK maternity unit. Observation was conducted on the postnatal ward and the newborn infant unit, and 30 mothers, 17 midwives, 4 neonatal nurses, 3 health care assistants, 3 senior house officers, and 3 senior pediatricians gave in-depth interviews during a 9-month period in 2002. One of the major themes that emerged was the cup-versus-bottle debate. There were 3 categories strongly linked to this theme: difficulties returning to the breast, ease of use, and necessary skills and knowledge. It appears there is an urgent need to determine which is the best method of giving supplementary feeds, so that full, accurate information can be given to mothers, appropriate policies be devised, and the necessary resources and staff training be provided. PMID- 15886342 TI - Breast pump adverse events: reports to the food and drug administration. AB - Breast pumps are medical devices used to express milk and maintain the milk supply. The purpose of this study was to characterize adverse events reported to the United States Food and Drug Administration (FDA) on breast pumps. Thirty seven adverse event reports on breast pumps were identified from the Manufacturer and User Facility Device Experience database between 1992 and 2003. Four additional reports were found in the Device Experience Network database from 1992 to 1996. The most commonly reported adverse events for electric breast pumps were pain, soreness, or discomfort; the need for medical intervention; and breast tissue damage. Most frequently reported problems for manual breast pumps were breast tissue damage and infection. Contamination of breast milk during pumping was also reported. Breast pump adverse events are likely underreported to the FDA. Reporting adverse events is important for improving the design and manufacture of breast pumps and subsequently decreasing adverse events. PMID- 15886343 TI - Extent, accuracy, and credibility of breastfeeding information on the Internet. AB - Our objective was to test and describe a model for evaluating Websites related to breastfeeding. Forty Websites most likely to be accessed by the public were evaluated for extent, accuracy, credibility, presentation, ease of use, and adherence to ethical and medical Internet publishing standards. Extent and accuracy of Website content were determined by a checklist of critical information. The majority of Websites reviewed provided accurate information and complied with the International Code of Marketing of Breast-milk Substitutes. Approximately half the Websites complied with standards of medical Internet publishing. While much information on breastfeeding on the Internet is accurate, there is wide variability in the extent of information, usability of Websites, and compliance with standards of medical Internet publishing. Results of this study may be helpful to health care professionals as a model for evaluating breastfeeding-related Websites and to highlight considerations when recommending or designing Websites. PMID- 15886344 TI - Manual expression of breast milk. PMID- 15886345 TI - Advocacy for breastfeeding: making a difference one community at a time. AB - In response to the need for health care professionals skilled in lactation management, a breastfeeding course was developed and taught at the University of Pennsylvania. Since 1995, Nursing 361 has been offered to undergraduate junior and senior students. The aim of this article is to discuss how through coursework, nursing students can provide breastfeeding advocacy and change the breastfeeding culture one community at a time. This article provides guidelines for others to conduct such projects, as well as exemplars to demonstrate how advocacy can change communities. Through development of an advocacy project during the course of a semester, a student learns about his or her identified community and is able to make an impact that often lasts years after his or her project is completed. PMID- 15886346 TI - Learning to lobby for probreastfeeding legislation: the story of a Texas bill to create a breastfeeding-friendly physician designation. AB - Legislation can reduce institutional barriers to breastfeeding. Lobbying is the process by which legislation is influenced by a special interest group. While generally thought of as an activity available only to the rich and powerful, lobbying by lactation activists can be an effective way to change public policy. A breastfeeding coalition in central Texas has been involved with lobbying efforts since 1995. Tactical lessons learned are shared to inspire and assist others seeking to pass probreastfeeding legislation. PMID- 15886347 TI - Managing your milk supply: going with the flow. PMID- 15886349 TI - Metabolism, excretion, and pharmacokinetics of (3-{[4-tert-butyl-benzyl) (pyridine-3-sulfonyl)-amino]-methyl}-phenoxy)-acetic acid, an EP2 receptor selective prostaglandin E2 agonist, in male and female Sprague-Dawley rats. AB - Metabolism, excretion, and pharmacokinetics of a highly selective EP2 agonist, CP 533,536 (3-{[4-tert-butyl-benzyl)-(pyridine-3-sulfonyl)-amino]-methyl}-phenoxy) acetic acid), were investigated in male and female Sprague-Dawley rats following an intravenous administration of a single 15 mg/kg dose of [(14)C]CP-533,536. At 144 h after the dose, the cumulative excretion of radioactivity averaged 98.2 +/- 3.44% and 97.0 +/- 4.82% in male and female rats, respectively. The radioactivity was predominantly excreted in feces, reaching 87% of the dose. Mean exposure [area under the concentration-time curve (AUC(0-infinity))] for both CP-533,536 and total radioactivity was higher in female rats than in male rats, whereas the plasma clearance of CP-533,536 and metabolites was lower in female rats compared to male rats. CP-533,536 was extensively metabolized in both male and female rats. The major oxidative pathway was due to the oxidation of the tert-butyl side chain to form the omega-hydroxy metabolite M4 (males, 19.7%; females, 6.5%). M4 was further oxidized to form the omega-carboxy metabolite M3 (males, 32.8%; females 1.66%) or conjugated via sulfation to form metabolite M6 (males 12.7%; females 36.2%). Other metabolites were due to N-oxidation of the pyridine ring (M5) and aromatic hydroxylation (M12), and conjugation with glucuronic acid. The secondary metabolites were due to N-dealkylation of the methyl-phenoxyacetic acid moiety and phase II conjugation. CP-536,536 accounted for about 63 and 72% of the AUC of the total radioactivity for male and female rats, respectively. Gender related differences in the metabolism and pharmacokinetics were observed. omega Carboxy metabolite M3 was the major metabolite in male rats, whereas M3-sulfate was identified as the major metabolite in female rats. PMID- 15886350 TI - Metabolism of verapamil in cultures of rat alveolar epithelial cells and pharmacokinetics after administration by intravenous and inhalation routes. AB - Administration of therapeutic entities by inhalation opens new possibilities for drug entry into systemic circulation, but this requires passage through the alveolar epithelium. Little is known about the pulmonary metabolism of verapamil. Specifically, this cardiovascular drug suffers from extensive first pass metabolism. We therefore evaluated the metabolism of verapamil in cultured alveolar epithelium and compared findings with results after administration by inhalation and intravenous routes. Specifically, cell culture of alveolar epithelium was characterized by gene expression of surfactant proteins A, B, C, and D, by immunohistochemistry of surfactant protein C, by staining for laminar bodies, and by gene expression of cytochrome P450 monooxygenases. During 6 days of culture expression, all cellular differentiation markers were obvious, albeit at different levels. With testosterone as substrate, we found alveolar epithelial cells to produce several stereo- and site-specific hydroxylation products. This provided evidence for metabolic competence of cultured alveolar epithelial cells. With verapamil as substrate, only limited production of metabolites was observed in cell culture assays, and similar results were recorded after administration by inhalation and intravenous routes. Likewise, elimination of verapamil from lung tissue and plasma was similar by both routes of administration. In conclusion, administration of verapamil by inhalation-abrogated extensive first pass metabolism frequently seen after oral application, and this may well be extended to the development of drugs with similar pharmacokinetic defects. PMID- 15886351 TI - Response of the ubiquitin-proteasome pathway to changes in muscle activity. AB - The ubiquitin-proteasome pathway plays a critical role in the adaptation of skeletal muscle to persistent decreases or increases in muscle activity. This article outlines the basics of pathway function and reviews what we know about pathway responses to altered muscle use. The ubiquitin-proteasome pathway regulates proteolysis in mammalian cells by attaching ubiquitin polymers to damaged proteins; this targets the protein for degradation via the 26S proteasome. The pathway is constitutively active in muscle and continually regulates protein turnover. Conditions of decreased muscle use, e.g., unloading, denervation, or immobilization, stimulate general pathway activity. This activity increase is caused by upregulation of regulatory components in the pathway and leads to accelerated proteolysis, resulting in net loss of muscle protein. Pathway activity is also increased in response to exercise, a two-phase response. An immediate increase in selective ubiquitin conjugation by constitutive pathway components contributes to exercise-stimulated signal transduction. Over hours-to days, exercise also stimulates a delayed increase in general ubiquitin conjugating activity by inducing expression of key components in the pathway. This increase mediates a late-phase rise in protein degradation that is required for muscle adaptation to exercise. Thus the ubiquitin-proteasome pathway functions as an essential mediator of muscle remodeling, both in atrophic states and exercise training. PMID- 15886352 TI - Adrenomedullin: angiogenesis and gene therapy. AB - Adrenomedullin (AM) is a potent, long-lasting vasodilator peptide that was originally isolated from human pheochromocytoma. AM signaling is of particular significance in endothelial cell biology since the peptide protects cells from apoptosis, promotes angiogenesis, and affects vascular tone and permeability. The angiogenic effect of AM is mediated by activation of Akt, mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2, and focal adhesion kinase in endothelial cells. Both AM and its receptor, calcitonin receptor-like receptor, are upregulated through a hypoxia-inducible factor-1-dependent pathway under hypoxic conditions. Thus AM signaling plays an important role in the regulation of angiogenesis in hypoxic conditions. Recently, we have developed a nonviral vector, gelatin. Positively charged gelatin holds negatively charged plasmid DNA in its lattice structure. DNA-gelatin complexes can delay gene degradation, leading to efficient gene transfer. Administration of AM DNA-gelatin complexes induces potent angiogenic effects in a rabbit model of hindlimb ischemia. Thus gelatin-mediated AM gene transfer may be a new therapeutic strategy for the treatment of tissue ischemia. Endothelial progenitor cells (EPCs) play an important role in endothelial regeneration. Interestingly, EPCs phagocytose ionically linked DNA-gelatin complexes in coculture, which allows nonviral gene transfer into EPCs. AM gene transfer into EPCs inhibits cell apoptosis and induces proliferation and migration, suggesting that AM gene transfer strengthens the therapeutic potential of EPCs. Intravenous administration of AM gene-modified EPCs regenerate pulmonary endothelium, resulting in improvement of pulmonary hypertension. These results suggest that in vivo and in vitro transfer of AM gene using gelatin may be applicable for intractable cardiovascular disease. PMID- 15886353 TI - Physiological regulation of food intake. PMID- 15886354 TI - A new way of looking at eating. PMID- 15886355 TI - A peripheral perspective of weight regain. PMID- 15886356 TI - Nandrolone decanoate modulates cell cycle regulation in functionally overloaded rat soleus muscle. AB - Functionally overloading rat soleus muscle by synergist ablation induces a rapid increase in mass. Muscle remodeling during the first week of overload is critical for the overload-induced growth. Anabolic steroid modulation of this overload induced remodeling response is not well understood. The purpose of this study was to determine whether pretreatment with nandrolone decanoate, a clinically administered anabolic steroid, alters muscle morphology and gene expression related to muscle growth during the initiation of functional overload in the rat soleus muscle. Adult (5 mo) male Fisher 344 x Brown Norway rats were randomly assigned to control (Sham), 3-day functional overload (OV), nandrolone decanoate administration (ND), or 3-day functional overload with nandrolone decanoate administration (OV+ND) treatment groups. Morphologically, OV increased the percentage of small (361%) and large (150%) fibers and expanded the ECM 50%. ND administration decreased the 3-day OV induction of small fibers 51% and nuclei associated with the ECM 20%. ND administration also attenuated the induction of cell cycle regulator p21 (64%) and myogenin (37%) mRNAs after 3 days of overload. These data demonstrate that nandrolone decanoate pretreatment can alter morphological and cell cycle regulator expression related to muscle growth at the onset of functional overload. PMID- 15886357 TI - Freezing tolerance of the European water frogs: the good, the bad, and the ugly. AB - Survival and some physiological responses to freezing were investigated in three European water frogs (Rana lessonae, Rana ridibunda, and their hybridogen Rana esculenta). The three species exhibited different survival times during freezing (from 10 h for R. lessonae to 20 h for R. ridibunda). The time courses of percent water frozen were similar; however, because of the huge differences in body mass among species (from 10 g for Rana lessonae to nearly 100 g for Rana ridibunda), the ice mass accumulation rate varied markedly (from 0.75 +/- 0.12 to 1.43 +/- 0.11 g ice/h, respectively) and was lowest in the terrestrial hibernator Rana lessonae. The hybrid Rana esculenta exhibited an intermediate response between the two parental species; furthermore, within-species correlation existed between body mass and ice mass accumulation rates, suggesting the occurrence of subpopulations in this species (0.84 +/- 0.08 g ice/h for small R. esculenta and 1.78 +/- 0.09 g ice/h for large ones). Biochemical analyses showed accumulation of blood glucose and lactate, liver glucose (originating from glycogen), and liver alanine in Rana lessonae and Rana esculenta but not in Rana ridibunda in response to freezing. The variation of freeze tolerance between these three closely related species could bring understanding to the physiological processes involved in the evolution of freeze tolerance in vertebrates. PMID- 15886358 TI - Exposure to chronic stress downregulates corticosterone responses to acute stressors. AB - We used captive European starlings (Sturnus vulgaris) to test whether corticosterone responses differed in birds held under normal laboratory conditions or conditions of chronic stress. Surprisingly, both basal corticosterone concentrations and corticosterone responses to acute stress were significantly reduced when birds were chronically stressed. To determine the mechanism underlying this reduced response, animals under both conditions were injected with lactated Ringer's solution (control), adrenocorticotropin (ACTH), arginine vasotocin (AVT), or dexamethasone (DEX). ACTH increased corticosterone concentrations above stress-induced levels in both cases, although maximum responses were lower in chronically stressed birds. AVT did not augment the corticosterone response under nonchronically stressed conditions, but it did under chronically stressed conditions. DEX reduced maximal corticosterone concentrations in both cases. Neither ovine nor rat corticotropin-releasing factor (CRF) altered normal stress responses. These data indicate that changes in responsiveness of the hypothalamic-pituitary-adrenal axis to ACTH and AVT serve to downregulate corticosterone responses during chronic stress. Furthermore, these data lead to the following hypothesis: ACTH output from the pituitary limits maximum corticosterone concentrations under normal conditions, but reduced AVT release from the hypothalamus regulates lower corticosterone concentrations under chronic stress conditions. PMID- 15886359 TI - N-acetyl-l-cysteine protects intestinal lymphocytes from apoptotic death after acute exercise in adrenalectomized mice. AB - Lymphocyte apoptosis has been observed after strenuous exercise. Both glucocorticoids (GC) and reactive oxygen species (ROS) have been suggested to contribute to exercise-induced lymphocyte apoptosis. The aims of this study were to 1) examine the direct contribution of GC during exercise-induced intestinal lymphocyte (IL) apoptosis and 2) determine the contribution of oxidative stress, in the absence of GC, to exercise-induced IL apoptosis. Mice were bilaterally adrenalectomized (ADX) and randomly assigned to receive saline (SAL) or N-acetyl l-cysteine (NAC) 30 min before treadmill exercise (EX). EX consisted of 90 min of continuous running at a 2 degrees slope (30 min at 22 m/min, 30 min at 25 m/min; and 30 min at 28 m/min), and then killed immediately (Imm) or 24 h (24 h) postexercise. Control mice were exposed to a nonexercised (NonEX) condition consisting of treadmill noise and vibration without running. ILs were isolated and measured for apoptotic (phosphatidylserine externalization, mitochondrial membrane depolarization, Bcl-2, caspase 3, and cytosolic cytochrome c) and oxidative stress (H(2)O(2) and glutathione) markers. Plasma was analyzed for corticosterone (CORT) by radioimmunoassay. ADX eliminated the exercise-induced elevation in CORT but did not prevent IL apoptosis and cell loss relative to NonEX mice. In contrast, administration of NAC to ADX mice protected ILs from apoptotic cell death and inhibited post-exercise cell loss. These findings suggest that GC are not responsible for exercise-induced apoptosis and cell loss of ILs. The protective effect provided by the antioxidant NAC strongly suggest that oxidative stress is the primary pathway for IL apoptosis and cell loss after strenuous exercise. PMID- 15886360 TI - Injection of neuropeptide W into paraventricular nucleus of hypothalamus increases food intake. AB - Neuropeptide W (NPW) is an endogenous ligand for G protein-coupled receptor 7 (GPR7). There are two forms of the peptide, designated as neuropeptide W-23 (NPW23) and neuropeptide W-30 (NPW30). In the current study we found that intracerebroventricular administration of NPW23 increased c-Fos immunoreactivity (IR) in a variety of brain sites, many of which are involved in the regulation of feeding. In particular, we noted that c-Fos IR levels were increased in hypocretin-expressing neurons in the perifornical region of the lateral hypothalamus (LH). We then studied whether injection of NPW23 into the paraventricular nucleus of the hypothalamus (PVN) and the LH increased food intake over a 24-h time period. Intra-PVN injection of NPW23 at doses ranging from 0.1 to 3 nmol increased feeding for up to 4 h, and doses ranging from 0.3 to 3 nmol increased feeding for up to 24 h. In contrast, only the 3-nmol dose of NPW23 increased feeding after administration into the LH. Together, these data suggest a modulatory role for NPW in the control of food intake. PMID- 15886361 TI - The cover. An Englishman in Moscow. PMID- 15886362 TI - A piece of my mind. Satisfaction? PMID- 15886363 TI - Gene variants explain patient differences in antiepileptic drug responses. PMID- 15886364 TI - The gut yields clues to obesity, therapies. PMID- 15886365 TI - Therapy takes wheeze out of cat allergies. PMID- 15886366 TI - In Europe, as in US, climate for embryo stem cell research is one of extremes. PMID- 15886367 TI - Officials track down pandemic flu samples. PMID- 15886371 TI - Consumption of vegetables and fruits and risk of breast cancer. PMID- 15886372 TI - Consumption of vegetables and fruits and risk of breast cancer. PMID- 15886373 TI - Fasting serum glucose level and cancer risk in Korean men and women. PMID- 15886374 TI - Pharmacological treatment of neuropsychiatric symptoms of dementia. PMID- 15886375 TI - Pharmacological treatment of neuropsychiatric symptoms of dementia. PMID- 15886376 TI - Antiretroviral treatment in pediatric HIV infection in the United States: from clinical trials to clinical practice. AB - CONTEXT: Antiretroviral therapy (ART) for pediatric human immunodeficiency virus (HIV) infection has evolved from simple nucleoside reverse transcriptase inhibitor (NRTI) regimens to complex combination therapies based largely on evidence from clinical trials. However, the integration of novel ART into the clinical care of pediatric HIV infection has not been examined. OBJECTIVES: To describe changes in the treatment of pediatric HIV infection in the United States from 1987-2003, to assess concordance of initial regimens with US pediatric guidelines, and to identify predictors of the first regimen switch. DESIGN, SETTING, AND PARTICIPANTS: The study population included 766 perinatally HIV infected children in the Pediatric AIDS Clinical Trials Group 219C cohort born before January 1, 2004, who had not participated in an ART clinical trial at 219C enrollment or during follow-up. MAIN OUTCOME MEASURES: Proportion of children receiving specific ART regimens, proportion of children initiating ART according to pediatric guidelines, and time to first regimen switch (risk of switching). RESULTS: Single and dual NRTI regimens were used most frequently through 1997. In 1998, 2 years after protease inhibitors were approved for adult HIV infection and at the time pediatric guidelines were issued, regimens of highly active antiretroviral therapy including a protease inhibitor became most frequently used. From 1998-2003, 22% of children initiated ART with a regimen not recommended by pediatric guidelines. In multivariate regression, the risk of switching decreased with age at ART initiation (hazard ratio [HR], 0.96; 95% confidence interval [CI], 0.94-0.99) and increased with year of initiation (HR, 1.28; 95% CI, 1.23-1.33). The risk of switching was higher in children who started with 1 NRTI (HR, 8.05; 95% CI, 5.80-11.18), 2 NRTIs (HR, 4.08; 95% CI, 3.08-5.40), or an unconventional regimen (HR, 6.23; 95% CI, 3.36-11.54) vs children who started with a protease inhibitor-containing regimen; and in children who initiated ART at CD4 T lymphocyte percentages less than 15 vs 15 or greater (HR, 2.90; 95% CI, 1.03-8.13). CONCLUSIONS: There was a short lag between the identification of novel ART and its adoption in the pediatric community. A variety of regimens were used, including some unorthodox therapies. Important predictors of first regimen switch were identified. PMID- 15886377 TI - Temporal trends in early clinical manifestations of perinatal HIV infection in a population-based cohort. AB - CONTEXT: The effect of early antiretroviral therapy (ART) on the early progression of perinatal human immunodeficiency virus (HIV) infection is not well defined. OBJECTIVE: To examine early disease progression and survival in a population-based cohort with perinatal HIV infection in relation to year of birth and use of ART. DESIGN, SETTING, AND PATIENTS: Retrospective study of temporal trends in early progression of perinatal HIV infection among 205 HIV-infected children in Northern California born between January 1, 1988, and December 31, 2001, and followed up through age 3 years. MAIN OUTCOME MEASURES: Prevalence of and age at progression to a first US Centers for Disease Control and Prevention category C diagnosis relative to year of birth, type of ART, and age at initiation of therapy. RESULTS: Of 205 children, 134 (65%) received ART and/or Pneumocystis jiroveci pneumonia prophylaxis. By age 3 years, 81 (40%) progressed to a category C diagnosis, 41 (51%) of whom died. Untreated children were significantly more likely to progress to a category C diagnosis (62% [44/71] untreated vs 28% [37/134] treated children, P<.001); none of 23 infants who received triple ART progressed to category C. However, even without triple ART, very early mono/dual ART (by age 2 months vs 3-4 months) was associated with delayed and decreased progression to category C (P = .02). Of 33 children born between January 1, 1996, and December 31, 2001, only 7 (21%) progressed to category C (P = .02 compared with 1988-1995), 6 of 7 of whom received no therapy. More recent year of birth and more advanced therapy were associated with improved survival. CONCLUSIONS: This population-based cohort demonstrated decreased early HIV progression and improved survival at age 3 years, associated with more advanced therapy. Although limited by small sample size, the findings suggest that very early treatment, even without triple ART, was associated with improved outcome. PMID- 15886378 TI - Follow-up testing among children with elevated screening blood lead levels. AB - CONTEXT: Follow-up testing after an abnormal screening blood lead level is a key component of lead poisoning prevention. OBJECTIVES: To measure the proportion of children with elevated screening lead levels who have follow-up testing and to determine factors associated with such care. DESIGN, SETTING, AND PARTICIPANTS: Retrospective, observational cohort study of 3682 Michigan Medicaid-enrolled children aged 6 years or younger who had a screening blood lead level of at least 10 microg/dL (0.48 micromol/L) between January 1, 2002, and June 30, 2003. MAIN OUTCOME MEASURE: Testing within 180 days of an elevated screening lead level. RESULTS: Follow-up testing was received by 53.9% (95% confidence interval [CI], 52.2%-55.5%) of the children. In multivariate analysis adjusting for age, screening blood lead level results, and local health department catchment area, the relative risk of follow-up testing was lower for Hispanic or nonwhite children than for white children (0.91; 95% CI, 0.87-0.94), for children living in urban compared with rural areas (0.92; 95% CI, 0.89-0.96), and for children living in high- compared with low-risk lead areas (0.94; 95% CI, 0.92-0.96). Among children who did not have follow-up testing, 58.6% (95% CI, 56.3%-61.0%) had at least 1 medical encounter in the 6-month period after the elevated screening blood lead level, including encounters for evaluation and management (39.3%; 95% CI, 36.9%-41.6%) or preventive care (13.2%; 95% CI, 11.6%-14.8%). CONCLUSIONS: The rate of follow-up testing after an abnormal screening blood lead level was low, and children with increased likelihood of lead poisoning were less likely to receive follow-up testing. At least half of the children had a missed opportunity for follow-up testing. The observed disparities of care may increase the burden of cognitive impairment among at-risk children. PMID- 15886379 TI - Association between phonocardiographic third and fourth heart sounds and objective measures of left ventricular function. AB - CONTEXT: The third (S3) and fourth (S4) heart sounds detected by phonocardiography are considered to represent the criterion standards of the gallop sounds, but their test characteristics have not been explored. OBJECTIVE: To determine the diagnostic test characteristics of the S3 and S4 for prediction of left ventricular dysfunction using a computerized heart sound detection algorithm. DESIGN, SETTING, AND PARTICIPANTS: Prospective study of 90 adult patients undergoing elective left-sided heart catheterization at a single US teaching hospital between August 2003 and June 2004. The mean age was 62 (SD, 13) years (range, 24-90 years) and 61 (68%) were male. Within a 4-hour period, participants underwent computerized heart sound phonocardiographic analysis, cardiac catheterization, transthoracic echocardiography, and blood sampling for assessment of an S3/S4, left ventricular end-diastolic pressure (LVEDP), left ventricular ejection fraction (LVEF), and B-type natriuretic peptide (BNP), respectively. MAIN OUTCOME MEASURES: Diagnostic test characteristics of the computerized phonocardiographic S3 and S4 using markers of left ventricular function as criterion standards. RESULTS: Mean (SD) LVEDP was significantly elevated (18.4 [6.9] mm Hg vs 12.1 [7.3] mm Hg; P<.001), mean (SD) LVEF was reduced (49.4% [20.2%] vs 63.6% [14.8%]; P<.001), and median (interquartile range) BNP was elevated (330 [98-1155] pg/mL vs 86 [41-192] pg/mL; P<.001) in those with an S3, S4, or both compared with patients without a diastolic heart sound. The sensitivities of these heart sounds to detect an elevated LVEDP, reduced LVEF, or elevated BNP were 41%, 52%, and 32% for an S3, and 46%, 43%, and 40% for an S4, respectively. For abnormal levels of the same markers of ventricular function, the specificities of the S3 were 92%, 87%, and 92%, while the specificities of the S4 were 80%, 72%, and 78%, respectively. CONCLUSIONS: Neither the phonocardiographic S3 nor the S4 is a sensitive marker of left ventricular dysfunction. The phonocardiographic S3 is specific for left ventricular dysfunction and appears to be superior to the moderate specificity of the phonocardiographic S4. PMID- 15886380 TI - Effects of a 5-lipoxygenase-activating protein inhibitor on biomarkers associated with risk of myocardial infarction: a randomized trial. AB - CONTEXT: Myocardial infarction (MI) is the leading cause of death in the world. Variants in the 5-lipoxygenase-activating protein (FLAP) gene are associated with risk of MI. OBJECTIVE: To determine the effect of an inhibitor of FLAP on levels of biomarkers associated with MI risk. DESIGN, SETTING, AND PATIENTS: A randomized, prospective, placebo-controlled, crossover trial of an inhibitor of FLAP (DG-031) in MI patients who carry at-risk variants in the FLAP gene or in the leukotriene A4 hydrolase gene. Of 268 patients screened, 191 were carriers of at-risk variants in FLAP (87%) or leukotriene A4 hydrolase (13%). Individuals were enrolled in April 2004 and were followed up by designated cardiologists from a university hospital in Iceland until September 2004. INTERVENTIONS: Patients were first randomized to receive 250 mg/d of DG-031, 500 mg/d of DG-031, 750 mg/d of DG-031, or placebo. After a 2-week washout period, patients received DG-031 if they had received placebo first or placebo if they had received DG-031 first. Treatment periods lasted for 4 weeks. MAIN OUTCOME MEASURES: Changes in levels of biomarkers associated with risk of MI. RESULTS: In response to 750 mg/d of DG 031, production of leukotriene B4 was significantly reduced by 26% (95% confidence interval [CI], 10%-39%; P = .003) and myeloperoxidase was significantly reduced by 12% (95% CI, 2%-21%; P = .02). The higher 2 doses of DG 031 produced a nonsignificant reduction in C-reactive protein (16%; 95% CI, -2% to 31%; P = .07) at 2 weeks. However, there was a more pronounced reduction (25%; 95% CI, 5%-40%; P = .02) in C-reactive protein at the end of the washout period that persisted for another 4 weeks thereafter. The FLAP inhibitor DG-031 was well tolerated and was not associated with any serious adverse events. CONCLUSION: In patients with specific at-risk variants of 2 genes in the leukotriene pathway, DG 031 led to significant and dose-dependent suppression of biomarkers that are associated with increased risk of MI events. PMID- 15886381 TI - Fracture prevention with vitamin D supplementation: a meta-analysis of randomized controlled trials. AB - CONTEXT: The role and dose of oral vitamin D supplementation in nonvertebral fracture prevention have not been well established. OBJECTIVE: To estimate the effectiveness of vitamin D supplementation in preventing hip and nonvertebral fractures in older persons. DATA SOURCES: A systematic review of English and non English articles using MEDLINE and the Cochrane Controlled Trials Register (1960 2005), and EMBASE (1991-2005). Additional studies were identified by contacting clinical experts and searching bibliographies and abstracts presented at the American Society for Bone and Mineral Research (1995-2004). Search terms included randomized controlled trial (RCT), controlled clinical trial, random allocation, double-blind method, cholecalciferol, ergocalciferol, 25-hydroxyvitamin D, fractures, humans, elderly, falls, and bone density. STUDY SELECTION: Only double blind RCTs of oral vitamin D supplementation (cholecalciferol, ergocalciferol) with or without calcium supplementation vs calcium supplementation or placebo in older persons (> or =60 years) that examined hip or nonvertebral fractures were included. DATA EXTRACTION: Independent extraction of articles by 2 authors using predefined data fields, including study quality indicators. DATA SYNTHESIS: All pooled analyses were based on random-effects models. Five RCTs for hip fracture (n = 9294) and 7 RCTs for nonvertebral fracture risk (n = 9820) met our inclusion criteria. All trials used cholecalciferol. Heterogeneity among studies for both hip and nonvertebral fracture prevention was observed, which disappeared after pooling RCTs with low-dose (400 IU/d) and higher-dose vitamin D (700-800 IU/d), separately. A vitamin D dose of 700 to 800 IU/d reduced the relative risk (RR) of hip fracture by 26% (3 RCTs with 5572 persons; pooled RR, 0.74; 95% confidence interval [CI], 0.61-0.88) and any nonvertebral fracture by 23% (5 RCTs with 6098 persons; pooled RR, 0.77; 95% CI, 0.68-0.87) vs calcium or placebo. No significant benefit was observed for RCTs with 400 IU/d vitamin D (2 RCTs with 3722 persons; pooled RR for hip fracture, 1.15; 95% CI, 0.88-1.50; and pooled RR for any nonvertebral fracture, 1.03; 95% CI, 0.86-1.24). CONCLUSIONS: Oral vitamin D supplementation between 700 to 800 IU/d appears to reduce the risk of hip and any nonvertebral fractures in ambulatory or institutionalized elderly persons. An oral vitamin D dose of 400 IU/d is not sufficient for fracture prevention. PMID- 15886382 TI - Palliative care in the final days of life: "they were expecting it at any time". AB - Providing care to actively dying patients presents unique challenges for the clinician. Patients in their final days require careful symptom management. Families need support and coaching as death approaches. Care does not end with the death of the patient but continues through death pronouncement, family notification of the death, discussion of autopsy, and immediate bereavement support. Skills in these and related areas are discussed in the context of one man's death, as illuminated by comments made after death by his daughter and his physician. This case also highlights the often differing perspectives of families and clinicians as they work to deal with a patient's death. A goal of mastering the palliative skills necessary to competently care for an actively dying patient is to enable a patient to die peacefully and relatively free of discomfort. Achieving such competency should also help to relieve clinician stress attendant to caring for dying patients. PMID- 15886383 TI - Balancing the upside and downside of antiretroviral therapy in children. PMID- 15886384 TI - Childhood lead poisoning prevention: too little, too late. PMID- 15886385 TI - Translating the Human Genome Project into prevention of myocardial infarction and stroke--getting close? PMID- 15886386 TI - JAMA patient page. Lead poisoning. PMID- 15886387 TI - Drosophila Wnt/Fz pathways. AB - Wnts [also known as Wingless (Wg)] are a family of conserved signaling molecules involved in a plethora of fundamental developmental and cell biological processes, such as cell proliferation, differentiation, and cell polarity. Dysregulation of the pathway can be detrimental, because several components are tumorigenic when mutated and are associated with hepatic, colorectal, breast, and skin cancers. First identified in the fruit fly Drosophila melanogaster as a gene family responsible for patterning the embryonic epidermis, the Wnt gene family, including Wg, encode secreted glycoproteins that activate receptor-mediated signaling pathways leading to numerous transcriptional and cellular responses. The main function of the canonical Wg pathway is to stabilize the cytoplasmic pool of a key mediator, beta-catenin [beta-catenin, known as Armadillo (Arm) in fruit flies], which is otherwise degraded by the proteasome pathway. Initially identified as a key player in stabilizing cell-cell adherens junctions, Arm is now known to also act as a transcription factor by forming a complex with the lymphoid enhancer factor (LEF)/T cell-specific transcription factor (TCF) family of high mobility group (HMG)-box transcription factors. Upon Wnt/Wg stimulation, stabilized Arm translocates to the nucleus, where, together with LEF/TCF transcription factors, it activates downstream target genes that regulate numerous cell biological processes. PMID- 15886388 TI - Text mining for metabolic pathways, signaling cascades, and protein networks. AB - The complexity of the information stored in databases and publications on metabolic and signaling pathways, the high throughput of experimental data, and the growing number of publications make it imperative to provide systems to help the researcher navigate through these interrelated information resources. Text mining methods have started to play a key role in the creation and maintenance of links between the information stored in biological databases and its original sources in the literature. These links will be extremely useful for database updating and curation, especially if a number of technical problems can be solved satisfactorily, including the identification of protein and gene names (entities in general) and the characterization of their types of interactions. The first generation of openly accessible text-mining systems, such as iHOP (Information Hyperlinked over Proteins), provides additional functions to facilitate the reconstruction of protein interaction networks, combine database and text information, and support the scientist in the formulation of novel hypotheses. The next challenge is the generation of comprehensive information regarding the general function of signaling pathways and protein interaction networks. PMID- 15886389 TI - A life science Semantic Web: are we there yet? AB - More than ever, life science researchers depend on information from multiple sources. The Semantic Web offers a powerful new strategy for consolidating both text and structured data into a comprehensive collections and views. In addition, these aggregates are readable by both humans and machines and could be the basis of information management and knowledge exchange. PMID- 15886390 TI - A mathematical and computational framework for quantitative comparison and integration of large-scale gene expression data. AB - Analysis of large-scale gene expression studies usually begins with gene clustering. A ubiquitous problem is that different algorithms applied to the same data inevitably give different results, and the differences are often substantial, involving a quarter or more of the genes analyzed. This raises a series of important but nettlesome questions: How are different clustering results related to each other and to the underlying data structure? Is one clustering objectively superior to another? Which differences, if any, are likely candidates to be biologically important? A systematic and quantitative way to address these questions is needed, together with an effective way to integrate and leverage expression results with other kinds of large-scale data and annotations. We developed a mathematical and computational framework to help quantify, compare, visualize and interactively mine clusterings. We show that by coupling confusion matrices with appropriate metrics (linear assignment and normalized mutual information scores), one can quantify and map differences between clusterings. A version of receiver operator characteristic analysis proved effective for quantifying and visualizing cluster quality and overlap. These methods, plus a flexible library of clustering algorithms, can be called from a new expandable set of software tools called CompClust 1.0 (http://woldlab.caltech.edu/compClust/). CompClust also makes it possible to relate expression clustering patterns to DNA sequence motif occurrences, protein DNA interaction measurements and various kinds of functional annotations. Test analyses used yeast cell cycle data and revealed data structure not obvious under all algorithms. These results were then integrated with transcription motif and global protein-DNA interaction data to identify G1 regulatory modules. PMID- 15886391 TI - An SOS-regulated operon involved in damage-inducible mutagenesis in Caulobacter crescentus. AB - DNA polymerases of the Y-family, such as Escherichia coli UmuC and DinB, are specialized enzymes induced by the SOS response, which bypass lesions allowing the continuation of DNA replication. umuDC orthologs are absent in Caulobacter crescentus and other bacteria, raising the question about the existence of SOS mutagenesis in these organisms. Here, we report that the C.crescentus dinB ortholog is not involved in damage-induced mutagenesis. However, an operon composed of two hypothetical genes and dnaE2, encoding a second copy of the catalytic subunit of Pol III, is damage inducible in a recA-dependent manner, and is responsible for most ultraviolet (UV) and mitomycin C-induced mutations in C.crescentus. The results demonstrate that the three genes are required for the error-prone processing of DNA lesions. The two hypothetical genes were named imuA and imuB, after inducible mutagenesis. ImuB is similar to proteins of the Y family of polymerases, and possibly cooperates with DnaE2 in lesion bypass. The mutations arising as a consequence of the activity of the imuAB dnaE2 operon are rather unusual for UV irradiation, including G:C to C:G transversions. PMID- 15886392 TI - Molecular haplotyping by linking emulsion PCR: analysis of paraoxonase 1 haplotypes and phenotypes. AB - Linking emulsion PCR (LE-PCR) enables formation of minichromosomes preserving phase information of two polymorphic loci, hence the haplotype. Emulsion PCR confines two amplicons of two linked polymorphic sites on a single template molecule to one aqueous-phase droplet. Linking PCR uses biotinylated, overlapping linking primers to connect these amplicons in the droplet. After LE-PCR, unlinked amplicons are removed on streptavidin-coated magnetic beads and single-stranded runoff products are capped by primer extension. Quantitative ASPCR can then be used to ascertain the haplotypes of the two polymorphic loci on the minichromosomes. Using LE-PCR, we determined the human paraoxonase-1 [PON1] molecular haplotypes at three loci (-909g>c, L55M, Q192R) in women who were compound heterozygotes for -909g>c/L55M (n = 89), -909g>c/Q192R (n = 77) and L55M/Q192R (n = 68). We observed a strong association between PON1 substrate specificity (paraoxon/phenylacetate substrate activity ratios) and -909g>c/Q192R haplotype. We have demonstrated here a powerful molecular haplotyping technology that can be applied in population studies. PMID- 15886393 TI - The highly conserved glutamic acid 791 of Rpb2 is involved in the binding of NTP and Mg(B) in the active center of human RNA polymerase II. AB - During transcription by RNA polymerase (RNAP) II, the incoming ribonucleoside triphosphate (NTP) enters the catalytic center in association with an Mg2+ ion, termed metal B [Mg(B)]. When bound to RNAP II, Mg(B) is coordinated by the beta and gamma phosphates of the NTP, Rpb1 residues D481 and D483 and Rpb2 residue D837. Rpb2 residue D837 is highly conserved across species. Notably, its neighboring residue, E836 (E791 in human RNAP II), is also highly conserved. To probe the role of E791 in transcription, we have affinity purified and characterized a human RNAP II mutant in which this residue was substituted for alanine. Our results indicate that the transcription activity of the Rpb2 E791A mutant is impaired at low NTP concentrations both in vitro and in vivo. They also revealed that both its NTP polymerization and transcript cleavage activities are decreased at low Mg concentrations. Because Rpb2 residue E791 appears to be located too far from the NTP-Mg(B) complex to make direct contact at either the entry (E) or addition (A) site, we propose alternative mechanisms by which this highly conserved residue participates in loading NTP-Mg(B) in the active site during transcription. PMID- 15886394 TI - Binding of pRNA to the N-terminal 14 amino acids of connector protein of bacteriophage phi29. AB - During assembly, bacterial virus phi29 utilizes a motor to insert genomic DNA into a preformed protein shell called the procapsid. The motor contains one twelve-subunit connector with a 3.6 nm central channel for DNA transportation, six viral-encoded RNA (packaging RNA or pRNA) and a protein, gp16, with unknown stoichiometry. Recent DNA-packaging models proposed that the 5-fold procapsid vertexes and 12-fold connector (or the hexameric pRNA ring) represented a symmetry mismatch enabling production of a force to drive a rotation motor to translocate and compress DNA. There was a discrepancy regarding the location of the foothold for the pRNA. One model [C. Chen and P. Guo (1997) J. Virol., 71, 3864-3871] suggested that the foothold for pRNA was the connector and that the pRNA-connector complex was part of the rotor. However, one other model suggested that the foothold for pRNA was the 5-fold vertex of the capsid protein and that pRNA was the stator. To elucidate the mechanism of phi29 DNA packaging, it is critical to confirm whether pRNA binds to the 5-fold vertex of the capsid protein or to the 12-fold symmetrical connector. Here, we used both purified connector and purified procapsid for binding studies with in vitro transcribed pRNA. Specific binding of pRNA to the connector in the procapsid was found by photoaffinity crosslinking. Removal of the N-terminal 14 amino acids of the gp10 protein by proteolytic cleavage resulted in undetectable binding of pRNA to either the connector or the procapsid, as investigated by agarose gel electrophoresis, SDS-PAGE, sucrose gradient sedimentation and N-terminal peptide sequencing. It is therefore concluded that pRNA bound to the 12-fold symmetrical connector to form a pRNA-connector complex and that the foothold for pRNA is the connector but not the capsid protein. PMID- 15886395 TI - The third helix of the homeodomain of paired class homeodomain proteins acts as a recognition helix both for DNA and protein interactions. AB - The transcription factor Pax6 is essential for the development of the eyes and the central nervous system of vertebrates and invertebrates. Pax6 contains two DNA-binding domains; an N-terminal paired domain and a centrally located homeodomain. We have previously shown that the vertebrate paired-less isoform of Pax6 (Pax6DeltaPD), and several other homeodomain proteins, interact with the full-length isoform of Pax6 enhancing Pax6-mediated transactivation from paired domain-DNA binding sites. By mutation analyses and molecular modeling we now demonstrate that, surprisingly, the recognition helix for specific DNA binding of the homeodomains of Pax6 and Chx10 interacts with the C-terminal RED subdomain of the paired domain of Pax6. Basic residues in the recognition helix and the N terminal arm of the homeodomain form an interaction surface that binds to an acidic patch involving residues in helices 1 and 2 of the RED subdomain. We used fluorescence resonance energy transfer assays to demonstrate such interactions between Pax6 molecules in the nuclei of living cells. Interestingly, two mutations in the homeodomain recognition helix, R57A and R58A, reduced protein protein interactions, but not DNA binding of Pax6DeltaPD. These findings suggest a critical role for the recognition helix and N-terminal arm of the paired class homeodomain in protein-protein interactions. PMID- 15886396 TI - DNA-tension dependence of restriction enzyme activity reveals mechanochemical properties of the reaction pathway. AB - Type II restriction endonucleases protect bacteria against phage infections by cleaving recognition sites on foreign double-stranded DNA (dsDNA) with extraordinary specificity. This capability arises primarily from large conformational changes in enzyme and/or DNA upon target sequence recognition. In order to elucidate the connection between the mechanics and the chemistry of DNA recognition and cleavage, we used a single-molecule approach to measure rate changes in the reaction pathway of EcoRV and BamHI as a function of DNA tension. We show that the induced-fit rate of EcoRV is strongly reduced by such tension. In contrast, BamHI is found to be insensitive, providing evidence that both substrate binding and hydrolysis are not influenced by this force. Based on these results, we propose a mechanochemical model of induced-fit reactions on DNA, allowing determination of induced-fit rates and DNA bend angles. Finally, for both enzymes a strongly decreased association rate is obtained on stretched DNA, presumably due to the absence of intradomain dissociation/re-association between non-specific sites (jumping). The obtained results should apply to many other DNA associated proteins. PMID- 15886397 TI - Genetic association, post-translational modification, and protein-protein interactions in Type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a complex disorder with a strong genetic component. Inherited complex disease susceptibility in humans is most commonly associated with single nucleotide polymorphisms. The mechanisms by which this occurs are still poorly understood. Here we focus on analyzing the effect of a set of disease-causing missense variations of the monogenetic form of Type 2 diabetes mellitus and a set of disease-associated nonsynonymous variations in comparison with that of nonsynonymous variations without any experimental evidence for association with any disease. Analysis of different properties such as evolutionary conservation status, solvent accessibility, secondary structure, etc. suggests that disease-causing variations are associated with extreme changes in the value of the parameters relating to evolutionary conservation and/or protein stability. Disease-associated variations are rather moderately conserved and have a milder effect on protein function and stability. The majority of the genes harboring these variations are clustered in or near the insulin signaling network. Most of these variations are identified as potential sites for post translational modifications; certain predictions have already reported experimental evidence. Overall our results indicate that Type 2 diabetes mellitus may result from a large number of single nucleotide polymorphisms that impair modular domain function and post-translational modifications involved in signaling. Our emphasis is more on conserved corresponding residues than the variation alone. We believe that the approach of considering a stretch of peptide sequence involving a polymorphism would be a better method of defining the role of the polymorphism in the manifestation of this disease. Because most of the variations associated with the disease are rare, we hypothesize that this disease is a "mosaic model" of interaction between a large number of rare alleles and a small number of common alleles along with the environment, which is little contrary to the existing common disease common variant model. PMID- 15886398 TI - Opposite effects of tumor necrosis factor and soluble fibronectin on junctional adhesion molecule-A in endothelial cells. AB - Junctional adhesion molecule-A (JAM-A) regulates key inflammatory responses, such as edema formation and leukocyte transmigration. Although it has been reported that the inflammatory cytokine tumor necrosis factor (TNF) causes the disassembly of JAM-A from the intercellular junctions, the mechanism has not been elucidated fully. Here, we report that TNF enhances the solubility of JAM-A in Triton X-100 and increases the amount of Triton-soluble JAM-A dimers at the cell surface but does not change the total levels of cellular JAM-A. Thus we hypothesized that TNF causes the redistribution of JAM-A from the junctions to the cell surface and that junction disassembly is sufficient to account for JAM-A redistribution. Intriguingly, however, even after complete disassembly of the junctions (with EDTA and trypsin), higher levels of JAM-A are detectable at the cell surface (by FACS analysis) in cells that had been previously incubated in the presence of TNF than in its absence. Thus we propose that TNF causes not only the disassembly of JAM-A from the junctions and its subsequent redistribution to the cell surface but also its dispersal in such a way that JAM-A becomes more easily accessible to the antibodies used for FACS analysis. Finally, we evaluated whether soluble fibronectin might attenuate the effects of TNF on JAM-A, as some inflammatory conditions are associated with the depletion of plasma fibronectin. We found that fibronectin reduces the effect of TNF on the disassembly of JAM-A, but not on its dispersal, thus further stressing that disassembly and dispersal can be functionally dissociated. PMID- 15886399 TI - Inhibition of surfactant activity by Pneumocystis carinii organisms and components in vitro. AB - This study examines the direct inhibitory effects of Pneumocystis carinii (Pc) organisms and chemical components on the surface activity and composition of whole calf lung surfactant (WLS) and calf lung surfactant extract (CLSE) in vitro. Incubation of WLS suspensions with intact Pc organisms (10(7) per milligram of surfactant phospholipid) did not significantly alter total phospholipid levels or surfactant protein A content. Incubation with intact Pc organisms also did not impair dynamic surface tension lowering in suspensions of WLS or centrifuged large surfactant aggregates on a bubble surfactometer (37 degrees C, 20 cycles/min, 0.5 and 2.5 mg phospholipid/ml). However, exposure of WLS or CLSE to disrupted (sonicated) Pc organisms led to severe detriments in activity, with minimum surface tensions of 17-19 mN/m vs. <1 mN/m for surfactants alone. Extracted hydrophobic chemical components from Pc (98.8% lipids, 0.1 mM) reduced the surface activity of WLS and CLSE similarly to sonicated Pc organisms, whereas extracted hydrophilic chemical components from Pc (primarily proteins) had only minor effects on surface tension lowering. These results indicate that in addition to surfactant dysfunction induced by inflammatory lung injury and edema-derived inhibitors in Pc pneumonia, disrupted Pc organisms in the alveolar lumen also have the potential to directly inhibit endogenous and exogenous lung surfactants in affected patients. PMID- 15886400 TI - Brain lateralization of emotional processing: historical roots and a future incorporating "dominance". AB - This article provides a review of research on the hemispheric specialization in emotional processing during the past 40 years and the theoretical models derived from the conceptual analysis of these results. The publications reviewed here were collected to better appreciate the cortical lateralization of emotional perception (visual and auditory), expression (facial and prosodic), and experience. Four major models of emotional processing are discussed--the Right Hemisphere, Valence, Approach-Withdrawal, and Behavioral Inhibition System Behavioral Activation System models. Observing the relative merits and limitations of these models, a new direction for exploration is offered. Specifically, to better appreciate the strength and direction (i.e., approach versus withdrawal) of experienced emotions, it is recommended that state "dominance" be evaluated in the context of asymmetrical activation of left frontal (dominance) versus right-frontal (submission) brain regions. PMID- 15886401 TI - The role of temporal structure in human vision. AB - Gestalt psychologists identified several stimulus properties thought to underlie visual grouping and figure/ground segmentation, and among those properties was common fate: the tendency to group together individual objects that move together in the same direction at the same speed. Recent years have witnessed an upsurge of interest in visual grouping based on other time-dependent sources of visual information, including synchronized changes in luminance, in motion direction, and in figure/ ground relations. These various sources of temporal grouping information can be subsumed under the rubric temporal structure. In this article, the authors review evidence bearing on the effectiveness of temporal structure in visual grouping. They start with an overview of evidence bearing on temporal acuity of human vision, covering studies dealing with temporal integration and temporal differentiation. They then summarize psychophysical studies dealing with figure/ground segregation based on temporal phase differences in deterministic and stochastic events. The authors conclude with a brief discussion of neurophysiological implications of these results. PMID- 15886402 TI - Effect of estrogen-serotonin interactions on mood and cognition. AB - Both the neurotransmitter serotonin and the ovarian steroid estrogen have been implicated in the modulation of mood and cognition. Although significant functional interactions between estrogen and serotonin are acknowledged, the nature of their relationship has not been fully elucidated. Research using ovariectomized animals has identified estrogen-induced changes in serotonin transmission, binding, and metabolism in brain regions implicated in the regulation of affect and cognition. Studies in humans, particularly of menopausal women undergoing estrogen treatment, have provided some support for these findings and identified instances in which change in mood or cognition is accompanied by alterations in serotonin function and hormonal status. However, it is apparent that further research is required to understand the neural processes involved in the interplay between estrogen and serotonin. By reviewing animal and human data regarding estrogen and serotonin's effects on mood and cognition, the authors aim to better define their relationship and highlight areas for further research. PMID- 15886404 TI - Lung function--clinical importance, problems, and new results. AB - This review tackles the usefulness of spirometry, a more than century old method of assessing pulmonary lung function. Variables measured with a spirometer, such as forced expiratory volume in 1 s, have long been the mainstays of the diagnosis and treatment of lung disorders. But there are problems with the reliability of spirometric measurements. The method depends on the cooperation of the investigated subject, which introduces a confounding subjective element and all too often results in test failure, and the results are evaluated against the predicted values that are based on a set of fixed factors, some of which, such as body height, are not in a straight proportion to the intrathoracic gas volume. Substantial spread of results arises, which makes a reliable assessment of lung function difficult. New methods, such as the resistance-volume curve, provide better information on airway behavior in different conditions. These new methods, which basically evolved from spirometry, show that the old idea of lung function analysis is still viable and may remain helpful for diagnosis and treatment of respiratory pathological states. PMID- 15886405 TI - Age-related response of the genioglossus muscle EMG-activity to hypoxia in humans. AB - The prevalence of snoring and obstructive sleep apnea syndrome increases with age. Upper airway obstruction during sleep occurs when the dilator muscles are unable to overcome the negative pharyngeal pressure. The very important muscle dilator of the upper airway is the genioglossus muscle (GG). Our previous study showed an increase in GG-EMG activity during progressive hypoxia. In the present study a hypoxic increase in GG-EMG activity in response to hypoxia was investigated in two groups of 10 healthy subjects each: aged 29.1 +/- and aged 53.2 +/-2.3. The older humans showed reduced response of GG to hypoxia. We suggest that the biological aging process may be a risk factor for the obstructive sleep apnea syndrome. PMID- 15886406 TI - Age-related peculiarities of breathing regulation and antioxidant enzymes under intermittent hypoxic training. AB - Aging is associated with changes in breathing regulation, particularly, in respiratory sensitivity to hypoxic stimuli. One theory of aging holds that reactive oxygen species play a key role in this process. These species have also been implicated in the carotid body O2 sensing. In the present study we investigated hypoxic ventilatory responses (HVR) and antioxidant enzymes activity in healthy young and elderly people under a 14-day adaptation to intermittent hypoxic training (IHT). The elderly demonstrated decreased HVR and blood catalase (CAT) activity on a background of strong negative correlation between the levels of end-tidal CO(2) tension and superoxide dismutase (SOD) activity. The adaptation to IHT resulted in increased HVR and SOD activity in both groups, and decreased CAT activity in young persons compared with its augmentation in the elderly. Increased SOD activity testifies to an overproduction of reactive oxygen species during IHT. We hypothesize that reactive oxygen species might potentiate a periodical augmentation of cytosolic Ca(2+) in the carotid body. This, in turn, might determine an increase in tyrosine hydroxylase gene expression and promote neurotransmitter synthesis/release, resulting in enhanced HVR. in the carotid body. This, in turn, might determine an increase in tyrosine hydroxylase gene expression and promote neurotransmitter synthesis/release, resulting in enhanced HVR. PMID- 15886407 TI - Cough sensitivity in localized scleroderma with no clinical symptoms from lower airways. AB - Cough sensitivity is increased in patients with atopic dermatitis, although they have no clinical symptoms from the lower airways. In the present study we examined the cough sensitivity to capsaicin in patients, who had no clinical respiratory symptoms, with sclerodermia localized to the skin. Cough sensitivity was defined as the lowest capsaicin concentration, which evokes 2 or more coughs. Twelve patients and 12 healthy matched volunteers, as a comparison group, inhaled deep breaths (2 L) of a capsaicin aerosol in doubled concentrations (from 0.02 to 200 micromol/L). Cough sensitivity, expressed as a geometric mean (95% CI) of capsaicin concentration, was 0.15 micromol/L (0.04 to 0.56) in the patients with localized sclerodermia and 4.96 micromol/L (2.50 to 9.85) in controls, which made a significant difference towards higher cough sensitivity in sclerodermia, respiratory symptom-free patients. Thus, disease processes localized outside the respiratory tract may have surreptitious pulmonary manifestation that is brought to light by the capsaicin cough test. PMID- 15886408 TI - Cortical activity during hypoxic hyperventilation. AB - This study seeks to determine the pattern of electroencephalogram changes during stimulatory ventilatory responses to acute progressive hypoxia. Electroencephalograms were recorded in the 10-20 electrode system during progressive poikilocapnic hypoxic tests based on the rebreathing routine. Healthy subjects were used for he study. A major finding was that hypoxia decreased the power spectra of the alpha activity. The decrease was surprisingly rapid and greater at mild hypoxic desaturation when pulmonary ventilation was about to pick up than during the maximum hypoxic hyperventilation. The possible relation of hypoxic decline in brain bioactivity to the manifestation of hypoxic hyperventilation remains to be elucidated in further studies. PMID- 15886409 TI - Intermittent hypoxia: mechanisms of action and some applications to bronchial asthma treatment. AB - Being essentially cut off from the global scientific community, Ukrainian and Russian scientists have developed a new concept for the beneficial use of adaptation to artificial intermittent hypoxia in treating of many diseases. The basic mechanisms underlying intermittent hypoxic training were elaborated mainly in three areas: regulation of respiration, free radical production and mitochondrial respiration. Twenty-year experience of the application of intermittent hypoxic therapy for the treatment of chronic obstructive bronchitis and bronchial asthma allows affirming that the adaptation to this kind of hypoxia causes a significant improvement of the clinical picture or even a complete recovery. The absence of negative side effects, typically observed during drug therapy, and the stimulation of organism's general, nonspecific resistance, makes the hypoxic therapy a treatment with a future. A special note is devoted to the use of intermittent hypoxic training in industrial health care for the purpose of prophylaxis and treatment of professional diseases. PMID- 15886410 TI - A pattern recognition method to distinguish gradual unilateral diaphragm paralysis in the cat. AB - This work deals with the application of a pattern recognition method to distinguish the degree of diaphragm paralysis after gradual unilateral sections of phrenic nerve rootlets in anesthetized, spontaneously breathing cats. The data set consisted of the features that characterize breathing pattern and of phrenic nerve amplitude. The method called for stratification of 6-dimensional vectors into three classes: intact, partial, and complete unilateral phrenicotomy, which offers the possibility to construe the classification rule on the basis of the information contained in a set of feature vectors with the known class membership. This method deals with the use of a distance function as a measure of similarity between two feature points. The results show that the degree of diaphragm paralysis could be recognized with the probability higher than 90%. Distinguishing the severity of diaphragmatic dysfunction and the compensatory strategies of the respiratory system, knowing only a handful of basic values describing breathing pattern, might have a practical meaning in respiratory emergencies. PMID- 15886411 TI - Role of nose breathing in genioglossus muscle response to hypoxia in older and younger subjects. AB - Several studies showed that nasal airway is an active component of the respiratory system. Clinical data suggest that nasal obstruction causes episodes of obstructive apnea or hypopnea. In the present study, a possible influence of breathing through the nose on genioglossus muscle (GG, dilator of upper airway) reactivity to hypoxic activation was studied. Two groups of 20-30 years and 41-55 years old, 35 healthy subjects each, were investigated. The GG-EMG-activity was recorded and analyzed during progressive normocapnic hypoxia. The subjects breathed through the nose (N) or mouth (M) alone and through the nose and mouth (N&M). Significantly smaller increases in the GG-EMG-activity in response to hypoxia were observed during M breathing, as compared with N and M&N breathing, in both groups. The older subjects also showed a reduced response of GG-muscle to hypoxia, which was most pronounced during M breathing. We suggest that breathing through the nose activates the dilator muscle of upper airways, preventing apnea events. PMID- 15886412 TI - Pharmacokinetics and metabolism of nicotine. AB - Nicotine (NIC), the major constituent of tobacco, is responsible for the compulsive use of tobacco. Advances in understanding of the pharmacokinetics and metabolism of NIC have been made rapidly over the past decade. The application of highly sensitive gas chromatography/mass spectrometry led to the identification and quantitation of new NIC metabolites as well as characterization of new pathways of NIC biotransformation. This review summarizes findings from human and animal studies concerning NIC kinetics and biotransformation as well as describes the factors that influence these processes. Recently, large individual, racial and species differences in the metabolism of NIC have been well documented. The differences in the metabolism of NIC may be a result of genetic, environmental, and developmental host influences. We review the scientific evidence from studies that supports a role for genetic mechanisms responsible for variability in the profile and the rate of the NIC metabolism. Actually, the majority of the genetic studies focus on the characterization of the CYP2A6 gene polymorphism, and on determining the relationship between the phenotype of NIC metabolism and the genotype of the CYP2A6 gene. There is good evidence that genetic polymorphisms associated with NIC metabolism are an important factor responsible for susceptibility to NIC dependence. It is anticipated that genetic findings can lead to the identification of individuals at a greater risk for tobacco addiction and will be used for more effective treatment and prevention strategies to reduce smoking. PMID- 15886413 TI - Stiripentol. A novel antiepileptic drug. AB - Epilepsy is one of the most widespread pathologies of human brain, affecting approximately 1% of world population. Despite the development of new methods of seizure control, chronic administration of antiepileptic drugs (AEDs) remains the treatment of choice. Nevertheless, pharmacotherapy is not always effective. In the case of single drug treatment, the number of non-responding patients is as high as 30%. Moreover, chronic medication with currently available AEDs may result in severe side-effects and undesired drug interactions. That is why in recent years intensive research has been carried out aiming at the development of new therapeutic strategies in epilepsy. The goal of this review is to assemble current literature data on stiripentol (STP), a novel anticonvulsant unrelated to any other AEDs. STP potentiates central gamma-aminobutyric acid (GABA) transmission and is characterized by nonlinear pharmacokinetics and inhibition of liver microsomal enzymes. STP has proved its anticonvulsant potency in different types of animal seizures, as well as in clinical trials. The drug seems a good candidate for adjunctive therapy in intractable epilepsy. PMID- 15886414 TI - Central effects of nafadotride, a dopamine D3 receptor antagonist, in rats. Comparison with haloperidol and clozapine. AB - The aim of this study was to examine behavioral and biochemical effects of nafadotride, the new dopamine D3 receptor antagonist, and to compare it with haloperidol (dopamine D2 receptor antagonist) and clozapine (predominate dopamine D4 receptor antagonist). Each drug was injected to adult male Wistar rats intraperitoneally, each at a single dose and for 14 consecutive days. Thirty minutes after single or last injection of the examined drugs, the following behavioral parameters were recorded: yawning, oral activity, locomotion, exploratory activity, catalepsy and coordination ability. By HPLC/ED methods, we determined the effects of the examined antagonists on the levels of biogenic amines in striatum and hippocampus: dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 3-methoxytyramine (3-MT), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) and noradrenaline (NA). Additionally, DA and 5-HT synthesis rate was determined in striatum and 5-HT in hippocampus. The results of the study indicate that nafadotride, the dopamine D3 receptor antagonist, has a behavioral and biochemical profile of action different from that of haloperidol but partially similar to that of clozapine. PMID- 15886415 TI - Anticonvulsant properties of N-(4-methylpiperazin-1-yl)- and N-[3-(4-methyl piperazin-1-yl)propyl] derivatives of 3-aryl- and 3-spirocycloalkyl-pyrrolidine 2,5-dione. AB - Two series of N-(4-methylpiperazin-1-yl)- and N-[3-(4-methylpiperazin-1-yl) propyl]-3-aryl- and 3-spirocycloalkyl-pyrrolidine-2,5-dione derivatives were synthesized and tested for anticonvulsant activity in the maximum electroshock (MES) seizure and pentetrazole (sc PTZ) seizure threshold tests. Compounds with an aromatic ring at position-3 of pyrrolidine-2,5-dione exhibited anticonvulsant activity in the MES test. For that series of compounds, ED50 values were determined. The most potent in the series were derivatives and with a chlorine atom at position-3 or 4 of the aromatic ring. Those compounds exhibited strong anticonvulsant activity, and their ED50 values ranged from 29 to 48 mg/kg. Introduction of the spirocycloalkyl ring into the position-3 of pyrrolidine-2,5 dione made those compounds inactive. PMID- 15886416 TI - Gingko biloba extract diminishes stress-induced memory deficits in rats. AB - Exposure to chronic restraint stress in rats and psychosocial stress in humans has been shown to alter cognitive functions such as learning and memory and has been linked to the pathophysiology of mood and anxiety disorders. Antianxiety or sedative agents used in the management of stress have several disadvantages and undesired effects. Therefore, in this study, we investigated efficacy of a natural medicine, the extract of Ginkgo biloba (EGB 761), in prevention and treatment of the post-stress memory dysfunctions. The results showed that chronic restraint stress (2 h for 21 days) or an 'equivalent' dose of exogenous corticosterone (5 mg/kg) impaired nonspatial memory as measured by an object recognition test. In control rats, EGB 761 improved spatial and nonspatial memory in Morris water maze and object recognition tests. Preventive doses of EGB 761 (100 mg/kg) normalized cognitive deficits, seen in rats chronically stressed or treated with corticosterone in object recognition test, and improved memory processes in these rats measured by Morris water maze test. There was no influence of our treatments on locomotor exploratory activity and anxiety measured in open field and elevated 'plus' maze tests, making a contribution of unspecific motor and emotional effects of the used drugs to their performance in the memory tests improbable. PMID- 15886417 TI - Histamine-induced cyclic AMP formation in the chick hypothalamus: interaction with vasoactive intestinal peptide. AB - Effects of histamine (HA) on cyclic AMP production and its action upon the effects evoked by vasoactive intestinal peptide (VIP) were studied in the chick hypothalamus. HA (0.1-1000 microM) potently stimulated cyclic AMP formation in the hypothalamic slices, reaching maximal effect (2.5-3.5-fold increase) at a 100 microM concentration, and displaying an EC50 value of approximately 6.5 microM/ The stimulatory action of HA was mimicked by agonists of HA receptors, with the following rank order of potency: HA>4-methylHA (H2)>or=Nalpha,Nalpha-dimethylHA (H3>>H1=H2)>or=2-methylHA (H1)>>amthamine (H2)>>dimaprit (H2) approximately tele methylHA. The HA (100 microM)-evoked increase in cyclic AMP production was concentration-dependently antagonized by selective H2-HA receptor blockers (aminopotentidine>>cimetidine>or=ranitidine>>zolantadine) and was not affected by mepyramine and thioperamide, a selective H1- and H3-HA receptor antagonist, respectively. The pharmacological profile of HA receptors linked to the cyclic AMP-generating system in the chick hypothalamus indicates that they represent either an avian-specific H2-like HA receptor or a novel subtype of HA receptors. Chicken VIP (cVIP; 0.1-3 microM) potently stimulated cyclic AMP synthesis in the chick hypothalamus in a concentration-dependent manner. A combination of cVIP with HA produced cyclic AMP response more than additive, and such a synergistic interaction was antagonized by ranitidine. It is suggested that in the avian brain HA and VIP may play in concert to regulate neuroendocrine processes. PMID- 15886418 TI - Immunosuppressive activity of an isoxazolo[5,4-e]triazepine--compound RM33. I. Effects on the humoral and cellular immune response in mice. AB - The aim of this investigation was to evaluate immunotropic properties of a new isoxazolotriazepine (compound RM33) in mice. We found that RM33 significantly inhibited induction of the humoral immune response to sheep red blood cells (SRBC), given 2 h prior to immunization. The development of the cellular immune response (delayed type hypersensitivity--DTH) to SRBC and to ovalbumin (OVA) was also suppressed, as well as the effector phase of the DTH to OVA. The compound was also effective when administered per os. The suppressive effects of RM33 on the immune response were comparable to those of cyclosporine A(CsA). We also showed that RM33 inhibited DTH to OVA if admixed with the sensitizing dose of an antigen and complete Freund's adjuvant (cFa) suggesting that the compound may affect initial events of antigen presentation. Such a hypothesis was supported by finding that RM33 significantly inhibited foot pad edema elicited by administration of cFa. The effects of RM33 on the activities of cytokines relevant to development and control of the immune response and inflammation such as: tumor necrosis factor alpha (TNF-alpha), interleukin 6 and 10 (IL-6 and IL 10) were also studied. The compound markedly (by 63%) inhibited lipopolysaccharide (LPS)-induced TNF-alpha serum level whereas IL-6 activity was lowered to a lesser extent (by 17%). The inducible IL-10 level in the splenocyte cultures was not affected at all. In summary, the presented results revealed immunosuppressive properties of RM33, which could be associated with its selective interference with co-stimulatory signals provided by adjuvant at initiation of the immune response. PMID- 15886419 TI - Effect of adminitration of etidronate and retinol on bone mechanical properties in ovariectomized rats. AB - The experiments were carried out on rats, divided into 7 groups: I-sham-operated control rats, II-ovariectomized (OVX) control rats, II- OVX+etidronate (10 mg/kg po), IV-OVX+retinol (700 IU/kg po), V-OVX+retinol (3,500 IU/kg po), VI OVX+etidronate (10 mg/kg po)+retinol (700 IU/kg po), VII-OVX+etidronate (10 mg/kg po)+retinol (3 500 IU/kg po). The drugs were administered once a day for 4 weeks. Bone mass, content of mineral substances and calcium were examined in the femur, tibia and L-4 vertebra. In the femur, mechanical properties of the whole bone supported on its epiphyses (the ultimate load, the breaking load and the deformation caused by the applied load) and of the femoral neck (the load causing the fracture) were studied. Bilateral ovariectomy induced unfavorable changes in mechanical properties of rat bones, which were partially prevented by administration of etidronate. Retinol at 3,500 IU/kg po caused intensification of the osteoporotic changes, especially it worsened the mechanical properties of bones. The results of the present study do not indicate the existence of any interaction between retinol and etidronate concerning mechanical properties of bones in OVX rats. PMID- 15886420 TI - Allyl disulfide as donor and cyanide as acceptor of sulfane sulfur in the mouse tissues. AB - Cytoplasm of mammalian glial cells was reported to contain Gomori-positive cytoplasmic granulation (GPCG), whose biological role is unknown. The present study attempted to discover conditions facilitating GPCG formation and to elucidate their relationship with sulfane sulfur metabolism. To address these problems, we investigated in vivo the effect of both allyl disulfide (DADS), occurring in garlic (sulfane sulfur donor) and cyanide (sulfane sulfur acceptor) on number of GPCG-containing glial cells in the mouse brain. In parallel, sulfane sulfur level and activity of rhodanese and 3-mercaptopyruvate sulfurtransferase (MpST) were determined in the mouse brain and liver. Cyanide caused a drop in GPCG number in the brain, while activity of sulfurtransferases and sulfane sulfur level remained unchanged. Slight but significant cyanide-induced rise in MpST activity was observed only in the liver, which indicates a possibility of enhancement of its detoxification in reaction with mercapropyruvate in this organ. DADS, a sulfur donor, increased GPCG number in the brain, whereas activity of sulfurtransferases and sulfane sulfur level did not change. However, in the liver, DADS elevated both sulfurtransferase activity and sulfane sulfur level. These observations suggest that DADS can constitute a source of sulfane sulfur for the liver, thereby activating anaerobic sulfur metabolism and sulfane sulfur transfer. Consequently, this leads to the increase in sulfane sulfur level in plasma, in which it is transported in the form of albumin hydropersulfides and can be used for cyanide detoxification or stored in glial cells as GPCG. Therefore, it is not excluded that GPCG observed in the brain of mice and other mammals can be a source and a store of sulfane sulfur in mammals. PMID- 15886421 TI - Antioxidant treatment normalizes renal Na+,K+-ATPase activity in leptin-treated rats. AB - Hyperleptinemia may be involved in the pathogenesis of obesity-associated hypertension, however, the mechanism of hypertensive effect of leptin is incompletely elucidated. Previously, we have demonstrated that chronic hyperleptinemia causes up-regulation of renal Na+,K+-ATPase and decreases urinary Na+ excretion. Herein, we investigated whether antioxidant treatment could correct these abnormalities. The study was performed on male Wistar rats. Leptin administered for 7 days (0.25 mg/kg twice daily sc) increased systolic blood pressure by 20.6%. Leptin had no effect on urine output and creatinine clearance but reduced sodium excretion by 40.1%. Na+,K+-ATPase activity in the renal cortex and medulla was higher in leptin-treated rats by 24.3% and 80.6%, respectively. In addition, hyperleptinemia was associated with an increase in plasma and urinary 8-isoprostanes and reduced urinary excretion of nitric oxide (NO) metabolites and cGMP. Co-treatment with a superoxide dismutase mimetic, tempol, or an NAD(P)H oxidase inhibitor, apocynin (2 mM in the drinking water), prevented leptin-induced blood pressure elevation, normalized plasma and urinary 8 isoprostanes, urinary excretion of sodium, NO metabolites and cGMP, as well as prevented up-regulation of renal Na+,K+-ATPase activity. These data suggest that hyperleptinemia increases renal Na+,K+-ATPase activity and reduces natriuresis by inducing oxidative stress-dependent NO deficiency. Antioxidant treatment is effective in leptin-induced hypertension and should be considered in controlling blood pressure in hyperleptinemic obese individuals. PMID- 15886422 TI - Synthesis and 5-HT1A/5-HT2A activity of some butyl analogs in the group of phenylpiperazine alkyl pyrimido[2,1-f]theophyllines. AB - New arylpiperazines with a four-methylene spacer containing a terminal pyrimido[2,1-f] theophylline fragment were synthesized, and their 5-HT1A and 5 HT2A receptor affinities were determined. All these compounds displayed a high affinity for 5-HT1A receptors (Ki=0.5-21.5 nM), and low affinity for 5-HT2A ones. The results of in vivo experiments showed that compounds revealed potential agonistic activity at presynaptic 5-HT1A receptors, whereas their functional activity at postsynaptic 5-HT1A sites was diversified. In fact, compounds and behaved like partial agonists, antagonists or agonists of postsynaptic 5-HT1A receptors, respectively. The pharmacological properties of the tested compounds were discussed in comparison with those of the three methylene-analogs described earlier. PMID- 15886423 TI - Isobolographic analysis of interactions among losigamone analog AO-620 and two conventional antiepileptic drugs. AB - In the present study, we investigated pharmacodynamic interactions among AO-620, a losigamone analog, and two conventional antiepileptic drugs, valproate (VPA) and phenobarbital (PB). Experiments were conducted in the maximal electroshock test in mice. Isobolographic analysis of the obtained data revealed pure additive interactions between AO-620 and PB applied at three dose ratios of 1:1, 1:3 and 3:1. Antagonism was observed when AO-620 was co-administered with VPAat the ratio of 3:1, while additive interactions were seen in two remaining proportions (1:3 and 1:1). Surprisingly, the interaction pattern of AO-620 appeared quite different from that of losigamone. PMID- 15886424 TI - MDR1 gene polymorphism in allogeenic kidney transplant patients with tremor. AB - P-glycoprotein (P-gp), an ATP-dependent efflux pump, is a membrane protein encoded by MDR1 gene, which demonstrates functional polymorphism. It is present in endothelial cells of the blood-brain barrier. P-gp pays a role in transmembrane transport of various xenobiotics, thus limiting their accumulation in the central nervous system. Cyclosporine A which is used as an immunosuppressive drug in patients with allogenic kidney grafts is a substrate for P-gp. Cyclosporine A may cause neurotoxic adverse effects, among them tremor. It was assumed that polymorphism of MDR1 gene which is associated with change in P-gp activity plays a role in induction of tremor in some patients with allogenic kidney graft treated with cyclosporine A. A total of 118 unrelated postransplant kidney patients were enrolled into the study. The tremor group included 23 cases and 95 randomly selected posttransplant individuals with no signs of tremor served as controls. No statistically significant correlation between MDR1 gene polymorphism C3435T and tremor was found. The tremor group and the control group were characterized by similar distribution of MDR1 genotypes, i.e. 3435CC, 3435CT, 3435TT. PMID- 15886426 TI - Evaluating the dynamic performance of a fibre optic pressure microsensor. AB - The dynamic performance of a new fibre optic sensor intended for measuring physiological fluid pressures is assessed in water. The sensor's sensitivity is evaluated at 23 degrees, 35 degrees and 37 degrees C against a Millar pressure catheter for sinusoidal pressure inputs with frequency ranging from 0.5 to 10 Hz. We found that sensitivity versus frequency is flat to 6 Hz and decreases slightly between 6 and 10 Hz. The sensitivity is slightly lower at 23 degrees C than at 37 degrees C. The reproducibility of measurements is excellent (two separate calibration tests in two consecutive days). The output of the fibre optic system used shows a constant time delay (0.13 s) for all frequencies tested. Experiments suggest that, with current sensor design, its immersion in degassed water prior to use ensures a reliable performance. PMID- 15886427 TI - Sodium fluorescein as a retinal pH indicator? AB - Retinal neovascularization is a symptom associated with various diseases revealing ocular fundus manifestation. Often, these neovascularizations originate from retinal hypoxia. A concomitant phenomenon of hypoxia is acidosis. To recognise this would permit the identification and treatment of hypoxic fundus areas long before first vascular modifications are seen. Thus, the goal of this investigation was to elucidate whether sodium fluorescein could be used as a retinal pH indicator. Sodium fluorescein solution was diluted in PBS (ratio: 1:150,000). The pH was varied from 6.5 to 8.6 by supplementation of HCl or NaOH, respectively. The fluorescence was excited by a pulsed diode laser (wavelength: 446 nm, pulse width: 100 ps) and detected by time-correlated single photon counting (TCSPC) technique. A least-squares fit of the measured fluorescence decay versus time by an exponential function results in the fluorescence lifetime. Ten measurements were taken at each pH for statistical analysis. The dependence of the fluorescence lifetime on the temperature and the concentration of sodium fluorescein was investigated in the same way. The fluorescence lifetime was found to rise from 3.775 ns to 4.11 ns with increasing pH (6.5 to 8.6). However, the gradient decreases with increasing pH. We found highly significant differences (Student's t-test, P<0.0005) of the fluorescence lifetimes for pH values with a mean difference of 0.125 at pH<7.65 whereas the differences were still significant (P7.65 and mean pH differences of 0.2. The fluorescence lifetime was independent of the temperature (22 degrees C to 37 degrees C) and the concentration of sodium fluorescein (dilution 1:150,000 to 1:2000). The fluorescence lifetime of sodium fluorescein depends on the pH but not on temperature and concentration. Thus, the discrimination of areas with retinal acidosis should be possible by combination of the TCSPC technique with scanning laser ophthalmoscopy. Further investigations have to clarify whether the accuracy of the measurement at the fundus in vivo is sufficient. PMID- 15886428 TI - Visualizing and manipulating individual protein molecules. AB - Single-molecule imaging and manipulation techniques have evolved in the past decade from mere jaw-dropping attractions to essential laboratory tools. By applying single-molecule methods important insights otherwise unavailable have been obtained on various biomolecular systems. Constantly improving single molecule imaging techniques keep expanding the scale of the explorable spatial detail, thereby providing possible solutions to getting around the debilitating diffraction limit present in physiological-condition structural investigations. In some areas, such as motor protein studies, single-molecule methods have become part of the routine and essential research toolkit. Entire research fields, such as single-molecule force spectroscopy, have been born. In the present review single-molecule visualization and manipulation methods are reviewed with a focus on proteins. Relevant signals and prominent applications are discussed along with experimental examples and recent important results. Finally, the perspectives of the single-molecule field are explored. PMID- 15886429 TI - Measurement for implantable rotary blood pumps. AB - Rotary blood pumps offer a cost-effective way to assist the failing heart. Relative to their pulsatile cousins, they can consist of remarkably few moving parts, with attendant advantages in reliability. These advantages are realized in full only if the entire assist system is kept maximally simple. Control of the pump must therefore be based on a minimum number of measurement devices. This paper reviews the measurements that are made in the wide range of implantable rotary blood pump designs that are in development for ventricular assist. In a number of these, fluid-mechanical variables are estimated indirectly from measurements of motor speed and current or power. The introduction explains the goals of rotary blood pump control by comparison to the innate properties of the natural heart. Then motor and fluid-mechanical variables that may be transduced are discussed. Methods of indirect estimation of pressure drop and flow-rate are dealt with, followed by ways of detecting unusual states such as inflow obstruction. It is found that detection of these alone can be the basis of an adequate control strategy. Some groups have estimated variables pertaining to the heart that is being assisted, and there has also been work on monitoring the ongoing health of the assist system itself. The review concludes with a brief look at the wider measurement context for the intensive-care facility that proposes to use such devices to provide circulatory support. PMID- 15886430 TI - Assessment of 1-lead and 2-lead electrode patterns in electrical impedance endotomography. AB - Electrical impedance endotomography (EIE) is a modality of impedance imaging where the electrodes are located on an insulating core placed at the centre of the region of interest. The absence of a physical limit to the medium surrounding the probe enables the use of remote electrodes. The present study compares the features of 2-lead measurements, where the two pairs of electrodes are located on the probe, to 1-lead measurements, where one of the two injection electrodes and one of the two sensing electrodes are located at a distance far away from the probe. The methodology was the characterization of the sensitivity matrix under the influence of electrode pattern, reconstruction radius and mesh construction. Three mesh constructions, three values of the reconstruction radius and five electrode patterns were compared. The study was carried out in 2D using calculated data. Measurement noise was simulated by an addition of 5% Gaussian white noise. The images were reconstructed using the Tikhonov method and L-curve technique. The results show that the reconstruction mesh and the radius of the reconstruction domain have less influence on the conditioning of the sensitivity matrix than the electrode pattern. Both 1-lead and 2-lead configurations enabled the reconstruction of images of relatively similar quality. Additional selection criteria are expected from hardware considerations. PMID- 15886431 TI - Double-wavelet approach to studying the modulation properties of nonstationary multimode dynamics. AB - On the basis of double-wavelet analysis, the paper proposes a method to study interactions in the form of frequency and amplitude modulation in nonstationary multimode data series. Special emphasis is given to the problem of quantifying the strength of modulation for a fast signal by a coexisting slower dynamics and to its physiological interpretation. Application of the approach is demonstrated for a number of model systems, including a model that generates chaotic dynamics. The approach is then applied to proximal tubular pressure data from rat nephrons in order to estimate the degree to which the myogenic dynamics of the afferent arteriole is modulated by the slower tubulo-glomerular dynamics. Our analysis reveals a significantly stronger interaction between the two mechanisms in spontaneously hypertensive rats than in normotensive rats. PMID- 15886432 TI - Leave-one-out prediction error of systolic arterial pressure time series under paced breathing. AB - In this paper, we consider systolic arterial pressure time series from healthy subjects and chronic heart failure patients, undergoing paced respiration, and show that different physiological states and pathological conditions may be characterized in terms of predictability of time series signals from the underlying biological system. We model time series by the regularized least squares approach and quantify predictability by the leave-one-out error. We find that the entrainment mechanism connected to paced breath, that renders the arterial blood pressure signal more regular and thus more predictable, is less effective in patients, and this effect correlates with the seriousness of the heart failure. Using a Gaussian kernel, so that all orders of nonlinearity are taken into account, the leave-one-out error separates controls from patients (probability less than 10(-7)), and alive patients from patients for whom cardiac death occurred (probability less than 0.01). PMID- 15886433 TI - Influence of data resolution and interpolation method on assessment of secondary brain insults in neurocritical care. AB - Continuous monitoring of physiologic vital signs is routine in neurocritical care. However, this patient information is usually only recorded intermittently (most often hourly) in the medical record. It is unclear whether this is sufficient to represent the occurrence of secondary brain insults (SBIs) or whether more frequent data collection will provide more comprehensive information for patient care. In 16 patients, physiologic data were acquired concurrently via two methods: per clinical routine, usually hourly, in the medical record (MR) and every minute via a custom data acquisition system (DA). SBIs were defined as a mean arterial pressure<90 mmHg, an intracranial pressure>20 mmHg or a temperature>37.5 degrees C. Number of events, cumulative duration of events and area under the curve (AUC) were compared between the two methods and 95% limits of agreement were assessed for various methods of MR data interpolation. For all three parameters, analysis of the DA and MR data frequently differed with regard to number of events, total duration of events and AUC. MR data tended to underestimate the number of total events. 95% limits of agreement were most narrow for trapezoidal interpolation of MR data, but even these limits were fairly broad. Assessment of secondary brain insults is highly dependent on (1) the temporal resolution of the method used to acquire patient data and on (2) the interpolation method if data are acquired intermittently. High frequency data acquisition may be necessary for more precise evaluation of secondary brain injury in neurocritical care. PMID- 15886434 TI - Three-block electrical model of renal impedance. AB - In this study, we measured the characteristic renal impedance profiles of Wistar rats and simulated the profiles using an electrical model with three series connected Windkessel blocks containing inductance. It is expected that a complete renal impedance profile ought to provide better physical properties information and have more diagnostic power than the pulsatility (PI) and resistive indices (RI) as a result of frequency dependency. A characteristic peak value at the third harmonic on the renal impedance amplitude curve was observed and the phase curve decreased with increasing harmonic numbers. From least mean square fitted parameters, the three blocks were given distinct physical properties and identified as: (1) the renal artery, (2) the small arteries plus the afferent arteriole and (3) the residual kidney (i.e., the efferent arteriole plus the post glomerular capillary structures). These allocations were made according to respective physical properties reported in previous research. These classifications were further confirmed when we compressed the kidney or infused Ang II. Variations in electrical parameters concurred with the likely affected blood vessels reported. This model describes renal impedance characteristics well; and it provides useful hints on the physical properties of the renal vascular system as well as allows for distinctions in possible physiologically affected locations during functional disturbance. It has potential for development as a clinical non-invasive diagnostic tool. PMID- 15886435 TI - A parametric model of the relationship between EIT and total lung volume. AB - Spirometry and electrical impedance tomography (EIT) data from 26 healthy subjects (14 males, 12 females) were used to develop a model linking contrast variations in EIT difference images to lung volume changes. Eight recordings, each 64 s long, were made for each subject in four postures (standing, sitting, reclining at 45 degrees, supine) and two breathing modes (quiet tidal and deep breathing). Age, gender and five anthropometric variables were recorded. The database was divided into four subsets. The first subset, data from 22 subjects (12 males, 10 females) recorded in deep breathing mode, was used to create the model. Validation was done with the other subsets: data recorded during quiet tidal breathing in the same 22 subjects, and data recorded in both breathing modes for the other four subjects. A quadratic equation in DeltaV(P) (lung volume changes recorded by the spirometer) provided a very good fit to total contrast changes in the EIT images. The model coefficients were found to depend on posture, gender, thoracic circumference and scapular skin fold. To validate the model, the quadratic equation was inverted to estimate lung volume changes from the EIT images. The estimated changes were then compared to the measured volume changes. Validations with each data subset yielded mean standard errors ranging from 9.3% to 12.4%. The proposed model is a first step in enabling inter individual comparisons of EIT images since: (1) it provides a framework for incorporating the effects of anthropometric variables, gender and posture, and (2) it references the images to a physical quantity (volume) verifiable by spirometry. PMID- 15886436 TI - Air mattress sensor system with balancing tube for unconstrained measurement of respiration and heart beat movements. AB - The cardio-respiratory signal is a fundamental vital sign used for assessment of a patient's status. Additionally, the cardio-respiratory signal provides a great deal of information to healthcare providers wishing to monitor healthy individuals. The air mattress sensor system allows the measurement of the respiration and heart beat movements without the use of a harness or sensor on the subject's body, which eliminates the difficulties these pose for long term measurements. In order to increase the sensitivity, a differential measurement technique between two air cells was used. The concept of a balancing tube between two air cells is suggested in order to increase the robustness against postural changes during the measurements. With this balancing tube, the meaningful frequency range could be selected using a pneumatic method. A mathematical model was constructed and validation experiments were performed for step and sinusoidal input signals. This technique was applied to measurements of respiration and heart beat movements in the supine posture on the bed, which showed potential for applications in sleep analysis, unconstrained healthcare monitoring and neonate monitoring. PMID- 15886437 TI - Cerebral microdialysis methodology--evaluation of 20 kDa and 100 kDa catheters. AB - Microdialysis monitoring of cerebral metabolism is now performed in several neuro intensive care units. Conventional microdialysis utilizes CMA70 catheters with 20 kDa molecular weight cut-off membranes enabling the measurement of small molecules such as glucose, lactate, pyruvate and glutamate. The CMA71 100 kDa molecular weight cut-off microdialysis catheter has recently been introduced to allow detection of larger molecules such as cytokines. The objective of this study was to perform in vitro and in vivo testing of the CMA71 microdialysis catheter, comparing its performance with the CMA70. In vitro comparison studies of three of each catheter using reference analyte solutions, demonstrated equivalent recovery for glucose, lactate, pyruvate and glutamate (range 94-97% for CMA70 and 88-103% for CMA71). In vivo comparison involved intracranial placement of paired CMA70 and CMA71 catheters (through the same cranial access device) in six patients with severe traumatic brain injury. Both catheters were perfused with CNS Perfusion Fluid without dextran at 0.3 microl min-1 with hourly sampling and bedside analysis on a CMA600 microdialysis analyser. The two catheters yielded equivalent results for glucose, lactate, pyruvate, glutamate and lactate/pyruvate ratio. CMA71 microdialysis catheters can, therefore, be used for routine clinical monitoring of extracellular substances, as well as for their intended research role of larger molecular weight protein sampling. PMID- 15886438 TI - Early recognition of regional cardiac ischemia using a 3-axis accelerometer sensor. AB - Perioperative mortality in coronary artery bypass grafting is usually caused by reduced left ventricular function due to regional myocardial ischemia or infarction. Post-operative graft occlusion is a well-known problem in coronary surgery. A sensitive tool to detect graft occlusion and monitor myocardial function may give the opportunity to revise malfunctioning grafts before departure from the hospital. This paper describes how a new method can detect cardiac ischemia using a 3-axis piezoelectric accelerometer. In three anesthetized pigs, a 3-axis piezoelectric accelerometer was sutured on the lateral free wall of the left ventricle. The left anterior descending (LAD) was occluded for different time periods and the accelerometer data were sampled with a PC. Short-time Fourier transform was calculated based on the accelerometer time series. The results were visualized using a 2D color-coded time-frequency plot. In the area of occlusion, a change to stronger power of higher harmonics was observed. Consequently, a difference value between the instant frequency pattern and a reference frequency pattern showed a rise in absolute value during the occlusion period. The preliminary results indicate that early recognition of regional cardiac ischemia is possible by analyzing accelerometer data acquired from the three animal trials using the prototype 3-axis accelerometer sensor. PMID- 15886439 TI - Low-cost oscillometric non-invasive blood pressure monitors: device repeatability and device differences. AB - Automated non-invasive blood pressure measuring devices based on the oscillometric technique are used widely for self-measurement and are often used in clinics in place of the manual, auscultatory method. Oscillometry was originally developed for monitoring purposes and there are questions over its suitability for making diagnostic measurements. This study measured the differences between automated devices, in the absence of physiological variability. We studied 19 low-cost, automated, non-invasive blood pressure devices, using a repeatable artificial arm simulator, and measured the within device repeatability and between-device differences. We found that the devices were repeatable (mean within-device difference 1 mmHg), but between-device differences were 4.4 mmHg (systolic pressure) and 3.6 mmHg (diastolic pressure), for normal and high-normal blood pressures. Individual devices are sufficiently repeatable for clinical trend use, but differences between devices are sufficiently large that they may be misinterpreted as clinically significant. PMID- 15886440 TI - Coordinated FA-MS and SIFT-MS analyses of breath following ingestion of D2O and ethanol: total body water, dispersal kinetics and ethanol metabolism. AB - A coordinated study of the dispersal of water between the various body compartments (stomach and gut, blood stream and tissue) and the similar dispersal kinetics of ethanol and its metabolism has been carried out involving two healthy volunteers using flowing afterglow mass spectrometry, FA-MS, and selected ion flow tube mass spectrometry, SIFT-MS. Thus, using these techniques, the variations of HDO and ethanol in breath, measured in successive single exhalations, were followed in real time after the ingestion of measured quantities of D2O and ethanol in proportion to the body weights of the subjects at the dose rates D2O approximately 0.283 g kg-1, ethanol approximately 0.067 g kg-1. During the FA-MS experimental periods (about 2 h), the dispersion of HDO into the body water and finally its equilibration in the total body water is observed from which total body water for each subject was determined. In the SIFT MS measurements, the dispersion of ethanol into the body water and its loss via metabolism was observed until the physiological (pre-dose) breath level of ethanol for each individual was restored. A simple linear transformation is used to derive the time variations of the blood levels of HDO and ethanol. This has allowed a comparison of the fractions of the ingested ethanol that are metabolized during first-pass metabolism for the two subjects. Thus, in one subject 30% and in the other subject 40% of the ingested alcohol is metabolized in the first 20 min following ingestion. The good time resolution allowed by non invasive breath analysis ensures that the rates of processes such as ethanol metabolism can be accurately measured. Simultaneous measurements of breath acetaldehyde (largely formed via the ethanol metabolism) and acetone were also performed during the SIFT-MS single breath exhalations. PMID- 15886441 TI - Automatic detection of cardiac waves on fetal magnetocardiographic signals. AB - Fetal magnetocardiography (fMCG) provides fetal cardiac traces useful for the prenatal monitoring of fetal heart function. In this paper, we describe an analytical model (ACWD) for the automatic detection of cardiac waves boundaries that works on fetal signals reconstructed from fMCG by means of independent component analysis. ACWD was validated for 45 healthy and 4 arrhythmic fetuses ranging from 22 to 37 weeks; ACWD outcomes were compared with the estimates of three independent investigators. Descriptive statistics were used to assess correspondence between the outcomes of the automatic and manual approaches. The parametric two-tailed Pearson correlation test (alpha=0.01) was employed to quantify, by means of the coefficients of determination, the amount of common variation between the sequences of intervals quantified automatically and manually. ACWD performances on short and long rhythm strips were investigated. ACWD demonstrated to be a robust tool providing dependable estimates of cardiac intervals and their variability during the third gestational trimester also in case of fetal arrhythmias. SNR and stability of fetal traces were the factors limiting ACWD performances. ACWD computation time, which was approximately 1:600 with respect to the manual procedure, was comparable with the time required for fCTI estimation on averaged beats. PMID- 15886442 TI - Characterization of volumetric flow rate waveforms in the normal internal carotid and vertebral arteries. AB - Knowledge of normal cerebrovascular volumetric flow rate (VFR) dynamics is of interest for establishing baselines, and for providing input data to cerebrovascular model studies. Retrospectively gated phase contrast magnetic resonance imaging was used to measure time-resolved VFR waveforms from the two internal carotid arteries (ICA) and two vertebral arteries (VA) of 17 young, normal volunteers (16M:1F) at rest in a supine posture. After normalizing each waveform to its respective cycle-averaged VFR, the timing and amplitude of feature points from the individual waveforms were averaged together to produce archetypal ICA and VA waveform shapes. Despite significant inter-individual differences in cycle-averaged VFR within the ICA compared to VA (275+/-52 versus 91+/-18 mL min-1), the respective waveform shapes were qualitatively similar overall. The VA waveform shape did, however, exhibit significantly higher amplitudes (e.g., peak:average VFR of 1.78+/-0.30 versus 1.66+/-0.16; p<0.05) and significantly higher variability both between and within subjects. A significant correlation was observed between peak and cycle-averaged VFR, suggesting that the representative waveform shapes presented here-when scaled by an individual's cycle-averaged VFR-may be used to characterize normal ICA and VA flow rate dynamics. This capability may be of particular utility for studies where cerebrovascular flow dynamics are required, but only average flow rates are available. PMID- 15886443 TI - Neural network based approach for anomaly detection in the lungs region by electrical impedance tomography. AB - In this paper, we have shown a simple procedure to detect anomalies in the lungs region by electrical impedance tomography. The main aim of the present study is to investigate the possibility of anomaly detection by using neural networks. Radial basis function neural networks are used as classifiers to classify the anomaly as belonging to the anterior or posterior region of the left lung or the right lung. The neural networks are trained and tested with the simulated data obtained by solving the mathematical model equation governing current flow through the simulated thoracic region. The equation solution and model simulation are done with FEMLAB. The effect of adding a higher number of neurons to the hidden layer can be clearly seen by the reduction in classification error. The study shows that there is interaction between the size (radius) and conductivity of anomalies and for some combination of these two factors the classification error of neural networks will be very small. PMID- 15886444 TI - Front-tracking image reconstruction algorithm for EIT-monitored cryosurgery using the boundary element method. AB - The effectiveness of cryosurgery, treatment of tumors by freezing, is highly dependent on knowledge of transient freezing extent, and therefore relies heavily on real-time imaging techniques for monitoring. Electrical impedance tomography (EIT) holds much promise for this application. In cryosurgery there is a three order of magnitude change in impedance across the freezing boundary and there is a priori knowledge of the freezing origin. Furthermore, an EIT image of the tissue can be done prior to the cryosurgery. In this study, we have developed an EIT front tracking reconstruction algorithm which takes advantage of these particular attributes of cryosurgery. The method tracks the freezing interface rather than the impedance distribution in the freezing tissue. In addition to drastically reducing the number of parameters needed to define the image, the computational complexity is further reduced by using the more appropriate boundary element method (BEM) for solution to the forward problem. The front tracking method was found to converge rapidly and accurately to a variety of simulated phantom images. PMID- 15886445 TI - Nonlinear characteristics of heart rate time series: influence of three recumbent positions in patients with mild or severe coronary artery disease. AB - The aim of this study was to investigate the nonlinear characteristics of heart rate variability (HRV) for three recumbent positions: the supine, left lateral and right lateral decubitus positions. Recently, using a linear analyses method (for time and frequency domains), the effect of the right lateral decubitus position on vagal modulation has been found to increase parasympathetic activity and decrease sympathetic modulation. Little is known about the nonlinear dynamics of HRV for the three recumbent positions. Therefore, we studied the correlation dimension (CD), the largest Lyapunov exponent (LLE), the sample entropy (SampEn), the approximate entropy (ApEn) and the exponent alpha of the 1/falpha power spectrum as nonlinear characteristics of HRV. In response to the right lateral decubitus position, the CD, LLE, SampEn and ApEn increased significantly in both coronary artery disease (CAD) and control groups. In the linear analyses, the normalized high-frequency power (nHF) increased in the right lateral decubitus position. The CD, LLE, ApEn and SampEn correlated positively to the nHF. The alpha exponent did not correlate to either linear measure or CD, but correlated negatively to LLE, ApEn and SampEn. Among the three recumbent positions, it was found that the right lateral decubitus position can increase the complexity of the human physiological system and the vagal modulation of the cardiac autonomic nervous system the most. PMID- 15886446 TI - Variability in time delay between two models of pulse oximeters for deriving the photoplethysmographic signals. AB - Pulse oximetry is commonly used as an arterial blood oxygen saturation (SaO2) measure. However, its other serial output, the photoplethysmography (PPG) signal, is not as well studied. Raw PPG signals can be used to estimate cardiovascular measures like pulse transit time (PTT) and possibly heart rate (HR). These timing related measurements are heavily dependent on the minimal variability in phase delay of the PPG signals. Masimo SET Rad-9 and Novametrix Oxypleth oximeters were investigated for their PPG phase characteristics on nine healthy adults. To facilitate comparison, PPG signals were acquired from fingers on the same hand in a random fashion. Results showed that mean PTT variations acquired from the Masimo oximeter (37.89 ms) were much greater than the Novametrix (5.66 ms). Documented evidence suggests that 1 ms variation in PTT is equivalent to 1 mmHg change in blood pressure. Moreover, the PTT trend derived from the Masimo oximeter can be mistaken as obstructive sleep apnoeas based on the known criteria. HR comparison was evaluated against estimates attained from an electrocardiogram (ECG). Novametrix differed from ECG by 0.71+/-0.58% (p<0.05) while Masimo differed by 4.51+/-3.66% (p>0.05). Modern oximeters can be attractive for their improved SaO2 measurement. However, using raw PPG signals obtained directly from these oximeters for timing-related measurements warrants further investigations. PMID- 15886447 TI - Gait information flow indicates complex motor dysfunction. AB - Gait-related back movements require coordination of multiple extremities including the flexible trunk. Ageing and chronic back pain influence these adjustments. These complex coordinations can advantageously be quantified by information theoretically based communication measures such as the gait information flow (GIF). Nine back pain patients (aged 61+/-10 yr) and 12 controls (aged 38+/-10 yr) were investigated during normal walking across a distance of 300 m. The back movements were measured as distances between characteristic points (cervical spine CS, thoracic spine TS, lumbar spine LS) by the sonoSens Monitor, a system for mobile motion analysis. Gait information flow and regularity indices (RI1: short prediction horizon of 100 ms, RI2: longer prediction horizon of walking period) were assessed as communication characteristics. All indices were non-parametrically tested for group differences. Sensitivity and specificity were assessed by bivariate logistic regression models. We found regularity indices systematically dependent on measurement points, information flow horizon and groups. In the patients RI1 was increased, but RI2 was decreased in comparison to the control group. These results quantitatively characterize the altered complex communication in the patients. We conclude that ageing and/or chronic back pain related dysfunctions of gait can advantageously be monitored by gait information flow characteristics of back movements measured as distances between characteristics points at the back surface. PMID- 15886448 TI - Springing the trap. PMID- 15886449 TI - Ion channels and D-amino acids. PMID- 15886450 TI - The economics of biotechnology. PMID- 15886451 TI - From mice to men: the evolution of the large, complex human brain. PMID- 15886452 TI - Living in a physical world II. The bio-ballistics of small projectiles. PMID- 15886453 TI - Ultraviolet irradiation initiates ectopic foot formation in regenerating hydra and promotes budding. AB - We have studied the effects of ultraviolet-C (UVC) and Ultraviolet-B (UVB) on growth and pattern formation in Pelmatohydra oligactis. UVC brings about a significant increase in budding in intact hydra while UVB does not exhibit such an effect. Excessive budding could be a response for survival at wavelengths that damage biological tissues. If the head or base piece of a bisected hydra is irradiated and recombined with the unirradiated missing part, regeneration proceeds normally indicating that exposure of a body part with either an intact head or foot to UVC does not influence pattern formation. Most significantly, in the middle piece, but not in the head or the base piece of a trisected hydra, UVC leads to initiation of ectopic feet formation in almost one third of the cases. Thus, UV irradiation interferes with pattern formation in regenerating hydra, possibly by changing positional values, and promotes budding in intact hydra. This is the first report on induction of ectopic feet formation by UV in regenerating hydra and opens up the possibility of using UV irradiation as a tool to understand pattern formation in the enigmatic hydra. PMID- 15886454 TI - In vitro translation of RNA to lactase during postnatal development of rat intestine. AB - mRNA levels encoding lactase were detected by Northern blot analysis using two different probes in developing rat intestine. Probe I and probe II corresponding to second half of prolactase gene showed a 6.8 kb mRNA transcript in 7, 14, 21 and 30 day old rat intestine. There was no change in quantity of lactase mRNA detected using probe II, but hybridization with probe I showed a progressive decrease in mRNA transcript encoding lactase with age. At day 7 and 14 of postnatal development, the lactase mRNA was quite high, but it reduced upon weaning. The in vitro translation products of RNA detected by Western blot analysis using brush border lactase antibodies showed several isoforms of lactase antigen with molecular weight ranging from 100-220 kDa. Analysed at days 7 and 30 of postnatal development, lactase isoforms of molecular weight 130 kDa and 220 kDa were similar to those found in purified brush border membranes. The translation of RNA to 220 kDa lactase protein was high in 7 and 14 day old pups, but it was markedly reduced in 30 day old animals. These findings support the contention that translation of mRNA to lactase is impaired in weaned animals, which may also be responsible for the maturational decline in lactase activity in adult rat intestine. PMID- 15886455 TI - Tissue localization and partial characterization of pheromone biosynthesis activating neuropeptide in Achaea janata. AB - Female sex pheromone production in certain moth species have been shown to be regulated by a cephalic endocrine peptidic factor: pheromone biosynthesis activating neuropeptide (PBAN), having 33 amino acid residues. Antisera against synthetic Heliothis zea-PBAN were developed. Using these polyclonals, immunoreactivity was mapped in the nervous system of Achaea janata. Three distinct groups of immunopositive secretory neurons were identified in the suboesophageal ganglion; and immunoreactivity was observed in the corpora cardiaca, thoracic and in the abdominal ganglia. From about 6000 brain sub oesophageal ganglion complexes, the neuropeptide was isolated; and purified sequentially by Sep-pak and reversed phase high performance liquid chromatographic methods. Identity of purified PBAN fraction was confirmed with polyclonal antibody by immunoblotting. Molecular mass of the isolated peptide was determined by matrix-assisted laser desorption/ionization mass spectrometry, and was found to be 3900 Da, same as that of known H. zea-PBAN. Radiochemical bioassay confirmed the pheromonotropic effect of the isolated neuropeptide in this insect. PMID- 15886456 TI - A non-polyene antifungal antibiotic from Streptomyces albidoflavus PU 23. AB - In all 312 actinomycete strains were isolated from water and soil samples from different regions. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising strain, Streptomyces albidoflavus PU 23 with strong antifungal activity against pathogenic fungi was selected for further studies. Antibiotic was extracted and purified from the isolate. Aspergillus spp. was most sensitive to the antibiotic followed by other molds and yeasts. The antibiotic was stable at different temperatures and pH tested and there was no significant loss of the antifungal activity after treatment with various detergents and enzymes. Synergistic effect was observed when the antibiotic was used in combination with hamycin. The antibiotic was fairly stable for a period of 12 months at 4 degree C. The mode of action of the antibiotic seems to be by binding to the ergosterol present in the fungal cell membrane resulting in the leakage of intracellular material and eventually death of the cell. The structure of the antibiotic was determined by elemental analysis and by ultraviolet (UV), Fourier transform infrared (FTIR), nuclear magnetic resonance (NMR) and liquid chromatography mass spectra (LCMS). The antibiotic was found to be a straight chain polyhydroxy, polyether, non-proteinic compound with a single double bond, indicating a nonpolyene antifungal antibiotic. PMID- 15886457 TI - cDNA cloning and characterization of a mannose-binding lectin from Zingiber officinale Roscoe (ginger) rhizomes. AB - Using RNA extracted from Zingiber officinale rhizomes and primers designed according to the conservative regions of monocot mannose-binding lectins, the full-length cDNA of Z. officinale agglutinin (ZOA) was cloned by rapid amplification of cDNA ends (RACE). The full-length cDNA of zoa was 746 bp and contained a 510 bp open reading frame (ORF) encoding a lectin precursor of 169 amino acids with a signal peptide. ZOA was a mannose-binding lectin with three typical mannose-binding sites (QDNY). Semi-quantitative RT-PCR analysis revealed that zoa expressed in all the tested tissues of Z. officinale including leaf, root and rhizome, suggesting it to be a constitutively expressing form. ZOA protein was successfully expressed in Escherichia coli with the molecular weight expected. To our knowledge, this is the first mannose-binding lectin cDNA cloned from the family Zingiberaceae. Our results demonstrate that monocot mannose binding lectins also occur within the family Zingiberaceae. PMID- 15886458 TI - Efficacy of lower doses of vanadium in restoring altered glucose metabolism and antioxidant status in diabetic rat lenses. AB - Vanadium compounds are potent in controlling elevated blood glucose levels in experimentally induced diabetes. However the toxicity associated with vanadium limits its role as therapeutic agent for diabetic treatment. A vanadium compound sodium orthovanadate (SOV) was given to alloxan-induced diabetic Wistar rats in lower doses in combination with Trigonella foenum graecum, a well-known hypoglycemic agent used in traditional Indian medicines. The effect of this combination was studied on lens morphology and glucose metabolism in diabetic rats. Lens, an insulin-independent tissue, was found severely affected in diabetes showing visual signs of cataract. Alterations in the activities of glucose metabolizing enzymes (hexokinase, aldose reductase, sorbitol dehydrogenase, glucose-6-phosphate dehydrogenase) and antioxidant enzymes (glutathione peroxidase, glutathione reductase) besides the levels of related metabolites, [sorbitol, fructose, glucose, thiobarbituric acid reactive species (TBARS) and reduced glutathione (GSH)] were observed in the lenses from diabetic rats and diabetic rats treated with insulin (2 IU/day), SOV (0.6 mg/ml), T. f. graecum seed powder (TSP, 5%) and TSP (5%) in combination with lowered dose of vanadium SOV (0.2 mg/ml), for a period of 3 weeks. The activity of the enzymes, hexokinase, aldose reductase and sorbitol dehydrogenase was significantly increased whereas the activity of glucose-6-phosphate dehydrogenase, glutathione peroxidase and glutathione reductase decreased significantly in lenses from 3 week diabetic rats. Significant increase in accumulation of metabolites, sorbitol, fructose, glucose was found in diabetic lenses. TBARS measure of peroxidation increased whereas the levels of antioxidant GSH decreased significantly in diabetic condition. Insulin restored the levels of altered enzyme activities and metabolites almost to control levels. Sodium orthovanadate (0.6 mg/ml) and Trigonella administered separately to diabetic animals could partially reverse the diabetic changes, metabolic and morphological, while vanadate in lowered dose in combination with Trigonella was found to be the most effective in restoring the altered lens metabolism and morphological appearance in diabetes. It may be concluded that vanadate at lowered doses administered in combination with Trigonella was the most effective in controlling the altered glucose metabolism and antioxidant status in diabetic lenses, these being significant factors involved in the development of diabetic complications, that reflects in the reduced lens opacity. PMID- 15886459 TI - Differential display-mediated identification of three drought-responsive expressed sequence tags in tea [Camellia sinensis (L.) O. Kuntze]. AB - There is no information on drought-modulated gene(s) in tea [Camellia sinensis (L.) O. Kuntze], a woody and perennial plant of commercial importance. Using differential display of mRNA, three drought-modulated expressed sequence tags (ESTs) were identified. Northern and BLAST analysis revealed that clone dr1 (drought-responsive), induced only by drought but not by ABA, showed significant scores with PR-5 (pathogenesis related) family of PR-protein gene. Another clone dr2, repressed by drought but not by ABA, had nucleotide repeats for polyasparate that are also present in chicken calsequestrin-like mRNA. Clone dr3, responded similarly to clone dr2 but did not show significant homology with the reported genes, hence appears to be novel. Identification of these ESTs is an initial step to clone the full length genes and their promoters. PMID- 15886460 TI - Differential cytotoxic effects of Annona squamosa seed extracts on human tumour cell lines: role of reactive oxygen species and glutathione. AB - Annonaceous acetogenins are a new class of compounds that have been reported to have potent pesticidal, parasiticidal, anti-microbial, cell growth inhibitory activities. In this study, organic and aqueous extracts from the defatted seeds of Annona squamosa (custard apple) were tested on different human tumour cell lines for antitumoural activity. While organic and aqueous extracts induced apoptosis in MCF-7 and K-562 cells, they failed to do so in COLO-205 cells. Treatment of MCF-7 and K-562 cells with organic and aqueous extracts resulted in nuclear condensation, DNA fragmentation, induction of reactive oxygen species (ROS) generation and reduced intracellular glutathione levels. In addition downregulation of Bcl-2 and PS externalization by Annexin-V staining suggested induction of apoptosis in MCF-7 and K-562 cells by both the extracts through oxidative stress. On the contrary, COLO-205 cells showed only PS externalization but no change in ROS and glutathione levels. These observations suggest that the induction of apoptosis by A. squamosa extracts can be selective for certain types of cancerous cells. PMID- 15886461 TI - Liver and kidney toxicity in chronic use of opioids: an experimental long term treatment model. AB - In this study, histopathological and biochemical changes due to chronic usage of morphine or tramadol in liver and kidney were assessed in rats. Thirty male Wistar rats (180-220 g) were included and divided into three groups. Normal saline (1 ml) was given intraperitoneally as placebo in the control group (n = 10). Morphine group (n = 10) received morphine intraperitoneally at a dose of 4, 8, 10 mg/kg/day in the first, second and the third ten days of the study, respectively. Tramadol group (n = 10), received the drug intraperitoneally at doses of 20, 40 and 80 mg/kg/day in the first, second and the third ten days of the study, respectively. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), creatinin, blood urea nitrogen (BUN) and malondialdehyde (MDA) levels were measured in the serum. Liver and kidney specimens were evaluated by light microscopy. Serum ALT, AST, LDH, BUN and creatinin levels were significantly higher in morphine group compared to the control group. Serum LDH, BUN and creatinin levels were significantly increased in the morphine group compared to the tramadol group. The mean MDA level was significantly higher in morphine group compared to the tramadol and control groups (P < 0.05). Light microscopy revealed severe centrolobular congestion and focal necrosis in the liver of morphine and tramadol groups, but perivenular necrosis was present only in the morphine group. The main histopathologic finding was vacuolization in tubular cells in morphine and tramadol groups. Our findings pointed out the risk of increased lipid peroxidation, hepatic and renal damage due to long term use of opioids, especially morphine. Although opioids are reported to be effective in pain management, their toxic effects should be kept in mind during chronic usage. PMID- 15886462 TI - Fingernails as biological indices of metal exposure. AB - Metal determination in human tissues is the most common application of biological monitoring for screening, diagnosis and assessment of metal exposures and their risks. Various biopsy-materials may be used. This paper deals with the quantitative determination of Cd, Pb, Cr, Mn, Fe, Ni, Cu, and Zn concentrations in nails of male subjects exposed to these metals along with their respective controls, while working in locomotive, carriage and road ways workshops, and lead battery factories. The levels of Cd, Pb, Cr, Mn, Fe, Ni, Cu and Zn in fingernails, assayed by atomic absorption spectrophotometry, were compared with their respective controls by student 't' test. All the obtained values were correlated to the personal and medical history of the subjects under study. Significantly high levels of Cd, Pb, Cr, Fe, Ni, Cu and Zn were present in smokers, compared to nonsmokers. The concentrations of Cd, Pb, Cr, Mn and Fe were not significantly high in vegetarian subjects. It was also observed that there is no contribution of liquor towards nail-metal concentration. Significant correlations were observed between skin disease and Cr, Mn, Fe, Cu; hypertension and Cd, Mn, Cu; mental stress and Cd, Pb, Mn, Ni, Cu, Zn; diabetes and Cr, Mn, Ni; chest pain and Pb; respiratory trouble and Cr, Mn, Fe, Ni, Zn; tuberculosis and Zn; acidity and Cd; and ophthalmic problems and Mn, Fe, Ni, and Zn. PMID- 15886463 TI - Evolutionary ecology in silico: Does mathematical modelling help in understanding 'generic' trends? AB - Motivated by the results of recent laboratory experiments, as well as many earlier field observations, that evolutionary changes can take place in ecosystems over relatively short ecological time scales, several 'unified' mathematical models of evolutionary ecology have been developed over the last few years with the aim of describing the statistical properties of data related to the evolution of ecosystems. Moreover, because of the availability of sufficiently fast computers, it has become possible to carry out detailed computer simulations of these models. For the sake of completeness and to put these recent developments in perspective, we begin with a brief summary of some older models of ecological phenomena and evolutionary processes. However, the main aim of this article is to review critically these 'unified' models, particularly those published in the physics literature, in simple language that makes the new theories accessible to a wider audience. PMID- 15886464 TI - Histone H1 modulates the antioxidant status in 9,10-dimethylbenz[a]anthracene induced experimental breast cancer. AB - AIM: Oxidative stress is implicated in the pathogenicity of cancer cells and contributes towards the response to antineoplastic agents. This study was aimed at assessing the antioxidant status in 9,10-dimethylbenz[a]anthracene-induced experimental animals treated with histone H1. METHODS: Histone H1 was injected and the antioxidant status was assessed in erythrocytes and tumour tissue of experimental animals. The antioxidant status was monitored from the levels of lipid peroxides as thiobarbituric acid reactants and conjugated dienes in the haemolysate, serum, glutathione, and plasma vitamins A, E, C and ceruloplasmin. In the haemolysate, activities of antioxidant enzymes, including superoxide dismutase, catalase, glutathione reductase, gamma-glutamyl transpeptidase, glutathione S-transferase, glutathione peroxidase and glucose-6-phosphate dehydrogenase, were assayed and erythrocyte fatty acid composition was analyzed by gas chromatography-mass spectrometry. All of the above parameters except fatty acid composition were assessed within the tumour tissue to assess the antioxidant status. RESULTS: Treatment with histone H1 enhanced the antioxidant in erythrocyte at the end of the 2nd and 4th week by significantly decreasing thiobarbituric acid-reactive substances and conjugated dienes, by increasing glutathione levels, activities of antioxidant enzymes and favourably altering the erythrocyte fatty acid composition. Within the tumour tissue, a significant atrophy with significant alteration in antioxidant and antioxidant enzyme status was evident in treatment. CONCLUSION: These results validate the role of histone H1 as an antitumour agent in breast cancer. PMID- 15886465 TI - Effects of cisplatin, 5-fluorouracil, and radiation on cell cycle regulation and apoptosis in the hypopharyngeal carcinoma cell line. AB - In head and neck cancer including hypopharyngeal cancer, cisplatin and 5 fluorouracil (5-FU) usually have been used as neoadjuvant chemotherapeutic agents. We investigated the effects of cisplatin, 5-FU and radiation on p53 protein expression and cell responses (cell cycle arrest and/or apoptosis) in the hypopharyngeal carcinoma cell line (PNUH-12; mutant type p53). PNUH-12 cells were treated with cisplatin, 5-FU and radiation. The changes in the cells were assessed by a cell cytotoxicity assay, Western blotting (p53 and p21(WAF1/CIP1) proteins), a DNA fragmentation assay, propidium iodide (PI) staining and DNA flow cytometry. The expression of p53 protein was increased after treatment with cisplatin and 5-FU, but not radiation. The expression of p21(WAF1/CIP1) protein was increased only after treatment with 5-FU, not cisplatin or radiation. With cisplatin and radiation, we observed apoptosis both by DNA fragmentation and PI staining and increased S phase in cisplatin and G2 phase in radiation by DNA flow cytometry. But, with 5-FU, we could not observe apoptosis by DNA fragmentation and PI stain but only an increased G1 phase by DNA flow cytometry. In PNUH-12, radiation induced p53-independent apoptosis and p21(WAF1/CIP1)-independent G2 phase cell cycle arrest. Cisplatin induced p53-dependent apoptosis and p21(WAF1/CIP1)-independent S-phase cell cycle arrest and 5-FU induced p53 and p21(WAF1/CIP1)-dependent G1-phase cell cycle arrest, not apoptosis. Cisplatin and 5-FU induced p53-dependent pathways, but radiation p53-independent pathway. The cell responses by cisplatin, 5-FU and radiation were all different pathways. PMID- 15886466 TI - Combined systemic chronotherapy and hepatic artery infusion for the treatment of metastatic colorectal cancer confined to the liver. AB - BACKGROUND: The optimal treatment of patients with metastatic colorectal cancer is still a clinical challenge. We describe the use of combined hepatic arterial infusion (HAI) of irinotecan (CPT-11) in conjunction with systemic chronotherapy infusion of 5-fluorouracil (5FU), folinic acid and carboplatin in patients with colorectal liver metastases. METHODS: Twenty-three patients with colorectal cancer and isolated liver metastases were enrolled in this trial. Intraoperative insertion of an intra-arterial catheter into the hepatic artery was accomplished during the colon operation (in cases of synchronous tumor) or as a separate procedure in colorectal cancer patients with newly diagnosed liver metastases. A systemic double-lumen double-chamber port was inserted via the subclavian vein as a separate procedure. The treatment plan included irinotecan given by intra arterial infusion at 150 mg/m2 for 1 h. After 2 weeks of rest chronomodulated 5FU (700 mg/m(2); peak delivery rate at 04:00 h), leucovorin (175 mg/m2; peak delivery rate at 04:00 h) and carboplatin (40 mg/m2; peak delivery rate at 16:00 h) for 4 days was followed by 10 days' rest and then given again. After 10 days' rest another HAI was introduced using the same method. Each cycle of therapy included 2 HAI courses and 2 chronotherapy courses in between. After 2 complete cycles, patients were evaluated for their response with weekly accessed toxicity recording. RESULTS: Seven women, 8 men, median age 61 years (range 46-72). Eight patients had synchronous colon and hepatic disease and 7 patients had metachronous disease. Ten patients had previously been treated with 5FU and leucovorin while 5 patients were chemonaive. The mean number of cycles were 11.6 per patient (range 8-19). Partial response was achieved in 6 patients (40%) and was followed by laparoscopic radiofrequency ablation in 5 patients (33%). Disease stabilization was observed in 2 patients (13%) and disease progression in 7 patients (47%) mainly after previous chemotherapy failure. Side effects were infrequent and mild including grade 2 GIT complaints (5 patients), RUQ pain during HAI (9 patients) and grade 2 hematological complaints in 2 patients. CONCLUSION: A combined chemotherapy protocol (HAI and chronotherapy) with irinotecan (CPT-11) together with chronomodulated infusion of 5FU, folinic acid and carboplatin can be used in metastatic colorectal patients with a high efficacy rate and minor side effects especially in pretreated patients. PMID- 15886467 TI - In vitro activity of linezolid against Staphylococcus aureus: a population study. AB - In our study, we aim to determine the existence of microorganisms that are heteroresistant to linezolid among Staphylococcus aureus clinical isolates in our setting between 1996 and 2002; during this period, linezolid was not used in clinical practice. There was no resistant subpopulation to 4 mg/l of linezolid in 99.4% of the strains. On the other hand, 16.46% of the strains exhibited resistant subpopulations to 4 microg/ml of vancomycin. However, the emergence of strains resistant to this drug has been described and the emergence of resistant strains should be monitored. PMID- 15886468 TI - Susceptibilities of Neisseria gonorrhoeae and Ureaplasma urealyticum isolates from male patients with urethritis to several antibiotics including telithromycin. AB - BACKGROUND: The minimal inhibitory concentrations (MICs) of erythromycin, azithromycin, clarithromycin, telithromycin, tetracycline, doxycycline, ciprofloxacin, ofloxacin, norfloxacin, levofloxacin, gemifloxacin and moxifloxacin against 78 Neisseria gonorrhoeae and 31 Ureaplasma urealyticum strains, which were isolated from patients with urethritis in Istanbul, were determined and compared. Additionally, the activities of penicillin and ceftriaxone against N. gonorrhoeae strains were explored. METHODS: MICs were determined by agar and broth dilution methods for N. gonorrhoeae and U. urealyticum, respectively. RESULTS: The susceptibility rates for penicillin and tetracycline in N. gonorrhoeae strains were 35.9 and 24.3%, respectively. All gonococcal strains were susceptible to ceftriaxone, with very low MICs (MIC90 0.008 microg/ml). Telithromycin was highly active against N. gonorrhoeae and U. urealyticum strains (MIC90 0.25 microg/ml for both). Ciprofloxacin was the most active quinolone against N. gonorrhoeae (MIC90 0.008 microg/ml) while quinolone resistance was detected in a single strain (1.3%). CONCLUSIONS: Tetracycline and penicillin should not be the option in empirical treatment of N. gonorrhoeae infections due to the very low susceptibility rates. Ceftriaxone continues to be the first choice antibiotic in the treatment of gonococcal urethritis. PMID- 15886469 TI - The irinotecan/5-fluorouracil combination induces apoptosis and enhances manganese superoxide dismutase activity in HT-29 human colon carcinoma cells. AB - BACKGROUND: We examined whether induction of apoptosis and Mn-superoxide dismutase (Mn-SOD) and Cu,Zn-superoxide dismutase (Cu,Zn-SOD) activities were involved in the greater cytotoxicity of the irinotecan (CPT-11)/5-fluorouracil (5 FU) combination for human colon cancer cells when compared to both drugs alone. METHODS: HT-29 and SNU-C4 human colon carcinoma cell lines were treated with 5-FU and CPT-11, then apoptosis was evaluated by flow cytometry and SOD activities were determined by polyacrylamide gel electrophoresis. RESULTS: Enhanced apoptosis of HT-29 cells was observed with all treatments containing 5-FU in SNU C4 cells; however, in HT-29 cells, apoptosis was enhanced only with the CPT-11/5 FU combination. In the SNU-C4 cell line, none of the treatments exerted a significant effect on Cu,Zn-SOD or Mn-SOD activity. However, in HT-29 cells, the CPT-11/5-FU combination enhanced Mn-SOD activity when compared to cells treated with CPT-11 alone. Nevertheless, the combined treatment did not interfere with Cu,Zn-SOD activity. CONCLUSION: Treatment with the CPT-11/5-FU combination may promote in HT-29 cell apoptosis by enhancing Mn-SOD activity. PMID- 15886470 TI - Tapering and discontinuation of glucocorticoid prophylaxis during prolonged weekly to biweekly paclitaxel administration. AB - BACKGROUND: Taxanes cause hypersensitivity reactions, averted by premedication with H1 blockers and high glucocorticoid (GC) doses. Prolonged weekly taxane administration may lead to GC toxicity. PURPOSE: To determine whether patients not hypersensitive to initial paclitaxel (PTX) infusion after high-dose GC premedication will tolerate subsequent, prolonged PTX treatment without GC prophylaxis. PATIENTS AND METHODS: In 115/122 breast cancer patients not hypersensitive to initial PTX treatment, 20 mg dexamethasone (DXM) doses were tapered by 2.0 mg/week, reaching 0 in those receiving 9 or more courses. After 4 PTX courses, diphenhydramine was administered orally, rather than intravenously. RESULTS: PTX was administered 143 times after 2.0-5.0 mg of DXM and 357 times without DXM. A total of 46 patients received 1-40 PTX courses without DXM. None of these 115 patients experienced hypersensitivity reactions. CONCLUSION: Patients unreactive to their first PTX infusions, after high-dose and tapering GC premedication, may not require GC prophylaxis for subsequent PTX therapy. PMID- 15886471 TI - A phase II study of docetaxel and infusional cisplatin in advanced non-small-cell lung cancer. AB - BACKGROUND: To evaluate the efficacy and safety of combination chemotherapy of cisplatin (5-day continuous infusion) and docetaxel for the treatment of previously untreated patients with advanced non-small-cell lung cancer (NSCLC). MATERIALS AND METHODS: Eligible patients had an ECOG performance status of 0-2 with measurable NSCLC. Patients received continuous infusion cisplatin 20 mg/m2/day on 5 days and bolus docetaxel 60 mg/m2/day (day 1; PiD therapy) at a 4 week interval. RESULTS: Forty-three patients were enrolled. The mean number of cycles administered per patient was 2, and ranged from 1 to 4. The response rate was 49% (95% confidence interval, 33.9-63.8%). The median survival time was 47 weeks and the 1-year survival rate was 47%. The major toxic effects were grade 3 or 4, neutropenia (88%), leukopenia (81%), thrombocytopenia (14%) and anemia (42%). There were no treatment-related deaths. CONCLUSION: PiD therapy was a well tolerated and active regimen for patients with advanced NSCLC. The major toxicity was neutropenia. PMID- 15886472 TI - Weekly paclitaxel in the treatment of advanced or metastatic breast cancer previously treated or not treated with docetaxel: a phase II study. AB - BACKGROUND: Paclitaxel has been approved for 3-weekly administration in Japan. Recent reports suggest that weekly paclitaxel can achieve a higher tumor response and lower toxicity. METHODS: This study was designed to investigate the usefulness and tolerability of weekly paclitaxel by 1-hour infusion in patients with metastatic breast cancer who were previously treated with docetaxel or other anticancer agents. RESULTS: Thirty-five patients were enrolled. The overall response rate was 41.2% (14/34, 95% confidence interval: 24.6-59.3%). The median time to progression and the median survival time were 218.5 and 624 days, respectively. One patient developed dyspnea, probably induced by a hypersensitivity reaction. The most common hematological toxicities were leukopenia and neutropenia, although no patients developed grade 4 leukopenia or neutropenia and G-CSF support was not required. CONCLUSIONS: Weekly paclitaxel could be safely administered and achieved a relatively high response rate. Weekly paclitaxel would be a good candidate second-line therapy for recurrent or advanced breast cancer. PMID- 15886473 TI - Phase I and pharmacokinetic evaluation of intravenous hyaluronic acid in combination with doxorubicin or 5-fluorouracil. AB - BACKGROUND: Pre-clinically, hyaluronan (HA) has been demonstrated to systemically target chemotherapeutic drugs to tumours while ameliorating treatment toxicities. This study is a preliminary clinical investigation to determine if HA could be safely used in combination with 5-fluorouracil (5-FU) and doxorubicin (DOX). METHODS: Thirty patients with metastatic cancer were intravenously administered 500 mg/m2 HA in combination with escalating doses of DOX (30-60 mg/m2) or 5-FU (cumulative dose of 1,350-2,250 mg/m2 per cycle). The effect of pre administration of 20 mg/m2 of folinic acid on HA/5-FU chemotherapy was also investigated. Patients were randomized to receive either HA/chemotherapy or chemotherapy alone in their first treatment cycle and vice versa for the second cycle. Patients received HA and chemotherapy in all subsequent cycles. RESULTS: Treatment was well tolerated, tumour responses were observed and the co administration of HA did not alter the pharmacokinetics of clinically relevant doses of 5-FU or DOX. CONCLUSION: High doses of intravenous high-molecular-weight HA can be safely co-administered with clinical doses of chemotherapy without significantly altering the toxicity or pharmacokinetics of the drugs or HA. PMID- 15886474 TI - A phase I study of prolonged ambulatory infusion of Ifosfamide with oral mesna. AB - BACKGROUND: Oral mesna allows investigation of ifosfamide as a prolonged ambulatory infusion for dose-intense out-patient use. METHODS: Cohorts of 3 patients received escalating doses of ifosfamide from 200 to 1,000 mg/m2/day as continuous ambulatory infusion with oral mesna at 30% of the ifosfamide dose every 6 h commencing 2 h prior to ifosfamide infusion as uroprotection on a 14 day schedule with cycles repeated every 28 days. RESULTS: Fifteen patients received a median of three cycles. Dose-limiting toxicities with cycle 1 were lethargy and hepatotoxicity at 1,000 mg/m2/14 days. Transient transaminase elevation was seen at all dose levels. The other grade 3 toxicities were single episodes of anaemia, granulocytopenia, nausea and hypotension. The best response was stable disease in a patient with thyroid cancer. CONCLUSION: Ambulatory infusion of 600 mg/m2 ifosfamide with 180 mg/m2 oral mesna was considered suitable for phase II trials and delivers dose-intense out-patient therapy without urotoxicity. PMID- 15886475 TI - Bacteremia in patients with hematological malignancies. AB - BACKGROUND: An appropriate regimen in the empirical therapy of neutropenic fever episodes must be individualized at each institution. Hospitals have different patterns of microbial isolates and antibiotic resistance that must be taken into account. The aim of this study was to investigate isolates of bacteremia and their antibiotic susceptibility in patients with hematological malignancies. METHODS: All positive blood cultures at a medical center in Taiwan between 1999 and 2002 from patients with hematological malignancies were evaluated. Eleven kinds of antibiotics were tested for antimicrobial activities. The risk factors for mortality were evaluated. RESULTS: Three hundred seventy-one episodes of bacteremia in 266 patients with hematological malignancies were recorded. Gram negative bacilli (GNB) were still predominant and accounted for 78.2% of isolates, followed by gram-positive cocci for 20.8% of isolates, and anaerobes for 1% of isolates. Escherichia coli was the most common isolated organism accounting for 27.5% of GNB isolates. Other isolates included Klebsiella pneumoniae (19.3%), Pseudomonas aeruginosa (11%), and Enterobacter cloacae (10.1%). The most isolated microorganisms were susceptible to cefoperazone/sulbactam, piperacillin/tazobactam, cefepime or imipenem. Age, GNB microorganism and inadequate empirical antibiotics were risk factors. CONCLUSIONS: We suggest that cefoperazone/sulbactam, piperacillin/tazobactam, cefepime or imipenem is an ideal empirical therapy. PMID- 15886476 TI - Phase I trial of intravenous cisplatin-topotecan chemotherapy for three consecutive days in patients with advanced solid tumors: parallel topotecan escalation in two fixed platinum dosing schemes. AB - PURPOSE: We performed a phase I study of two fixed dosing schemes of cisplatin, a DNA cross-linker, with intravenous escalating topotecan, a DNA-topoisomerase I inhibitor. EXPERIMENTAL DESIGN: 40 patients with advanced solid tumors received intravenous cisplatin at a fixed dose of either 25 mg/m2 (schedule A) or 20 mg/m2 (schedule B) daily for 3 days with standard hydration. Topotecan escalation proceeded in 0.75, 0.90, 1.0, 1.15 mg/m2 cohorts in schedule A and 1.0, 1.1, 1.2, 1.3 mg/m2 cohorts in schedule B, administered intravenously at the end of cisplatin infusion daily for 3 days, repeated every 3 weeks. Dose-limiting toxicity (DLT) consisted of protracted grade IV neutropenia, febrile neutropenia, grade IV thrombocytopenia and any grade III/IV non-hematological toxicity. Epoetin and granulocyte colony-stimulating factor support was allowed on severe myeloablation. Endpoints were the identification of maximal tolerated dose (MTD), DLT and other toxicity. RESULTS: The MTD was reached in cohort 25/1.15 mg/m2 in schedule A and 20/1.2 mg/m2 in schedule B. All DLT seen consisted of three episodes of febrile neutropenia and two of grade IV thrombocytopenia in schedule A, with three episodes of febrile neutropenia and one of protracted neutropenia in schedule B. Myelosuppression was substantial in all cohorts despite granulocyte colony-stimulating factor and epoetin support, peaked on the third week of treatment and resulted in administration of chemotherapy at a median of every 4 weeks. Non-hematologic toxicity was mild. The response rate was 51% with seven complete responses occurring in patients with ovarian cancer, small cell and non-small cell lung cancer and cancer of unknown primary. The recommended dose was 20/ 1.1 mg/m2 for cisplatin and topotecan on schedule B, as the number of responses and administered topotecan dose were higher in schedule B recommended dose with lower cisplatin dose, minimizing problems of nephrotoxicity and vomiting. CONCLUSIONS: The schedule B daily cisplatin-topotecan x 3 combination with secondary cytokine support is associated with promising activity and schedule convenience. However, substantial myelosuppression undermines its applicability in the palliative setting, stressing the need for less toxic regimens. PMID- 15886477 TI - Fosfomycin tromethamine in uncomplicated urinary tract infections: a clinical study. AB - The aim of our study was to verify if the empiric therapy with a single dose of 3 g fosfomycin tromethamine in patients with uncomplicated urinary tract infections (UTIs) was able to clinically resolveand to microbiologically eradicate the infection. A total of 387 out of the 400 patients (274 cases with acute and 113 cases with recurrent uncomplicated UTIs) were enrolled in the clinical study. Clinical and microbiological assessments were performed before and at 8-10 days after the administration. At follow-up high clinical recovery (88.9%) and bacteriological (94.9%) eradication rates were achieved. Gastrointestinal side effects were found in only 4.3% of patients. In conclusion, a single-dose administration regimen of fosfomycin tromethamine should be encouraged as a first choice of drug therapy for uncomplicated UTIs. PMID- 15886478 TI - Exacerbation of psoriasis due to peginterferon alpha-2b plus ribavirin treatment of chronic active hepatitis C. AB - A 45-year-old woman with chronic hepatitis C was treated with peginterferon alpha 2b in combination with ribavirin, which activated psoriasis. In the patient, psoriasis reoccurred after being inactive for 10 years. One week after institution of therapy, she presented with plaques of psoriasis at her injection site having been administered a first dose of peginterferon alpha-2b. Psoriatic lesions of the patient worsened dramatically during therapy. Cutaneous lesions appeared at the injection sites, in the face, the scalp hair, at the back of the ears, under the breasts and armpits, and in the regions of the anus and elbows. In the second week of treatment, transaminase levels of the patient regressed to normal. Detectable hepatitis C virus RNA in the serum was eliminated at week 14 of treatment. Because of the severity of psoriatic disease activity, therapy was discontinued at the end of 14 weeks after treatment onset. This side effect of peginterferon alpha-2b plus ribavirin treatment should be kept in mind in the treatment of patients with a history of psoriasis combined with active hepatitis C. PMID- 15886479 TI - Postnatal expression of the serotonin transporter in auditory brainstem neurons. AB - To investigate the putative role of serotonin (5-HT) in auditory brainstem development, the expression of the 5-HT transporter (5-HTT) was evaluated in the normal mouse brainstem at 6 different postnatal ages. The brains of C3H/HeJ mice at birth (P0) and P1, P8-P9, P13, P21-P22, P35-P36 and P48-P50 were collected and processed immunohistochemically with an antibody raised against the 5-HTT. 5-HTT immunoreactivity (5-HTT-IR) was first observed in P8 mice and was localized to cell bodies in the ventral cochlear nucleus (VCN) and principal nuclei of the superior olivary complex, including the medial nucleus of the trapezoid body. Labeled neurons were found in similar regions in older mice except at P48-50, where labeled neurons were observed in the VCN only. 5-HTT-IR was especially prominent in VCN neurons at P21 and was observed in all of the brains examined at this age. These results indicate that auditory brainstem neurons of the normal inbred mouse express the 5-HTT postnatally. The presence of 5-HTT-IR in neurons located in the VCN indicates a regional expression of the 5-HTT that is related to the ascending auditory pathway. The timing of 5-HTT expression indicates that 5-HT may modulate developmental processes that rely on cochlear input. PMID- 15886480 TI - Peptide-mediated protection from ethanol-induced neural tube defects. AB - Ethanol inhibition of L1-mediated cell adhesion may contribute to the spectrum of neurological, behavioral and morphological abnormalities associated with prenatal ethanol exposure. We showed previously that the neuroprotective peptides NAPVSIPQ (NAP) and SALLRSIPA (SAL) antagonize ethanol inhibition of L1 adhesion and prevent ethanol-induced growth retardation in mouse whole embryo culture. Here we ask whether NAP and SAL also prevent ethanol-induced major malformations of the nervous system. Gestational day 8.0 (3-5 somites) C57BL/6J mouse embryos were grown for 6 h in control medium, 100 mM ethanol and 10(-10) M peptides and then maintained for an additional 20 h in control medium. At the end of the culture period, only embryos having 18-19 somite pairs were examined and compared for the degree of neural tube closure. Ethanol exposure resulted in neural tube defects (NTDs) consistent with total dysraphia and anencephaly. Co-incubation with ethanol and L-NAP (all L-amino acids), D-NAP (all D-amino acids) or SAL significantly increased the percentage of embryos that had begun to close their neural folds at the level of the forebrain/midbrain junction or that had progressed beyond this stage of closure. P7A-NAP (NAPVSIAQ), which lacks neuroprotective activity, but retains activity as an antagonist of ethanol inhibition of L1 adhesion, was effective in preventing ethanol-induced NTDs. In contrast, I6A-NAP (NAPVSAPQ), which shows reduced efficacy as an ethanol antagonist but retains its neuroprotective efficacy, did not significantly diminish the induction of NTDs by ethanol. These findings demonstrate the ability of NAP and SAL to prevent ethanol-induced NTDs and support the hypothesis that ethanol teratogenesis is caused in part by ethanol inhibition of L1-mediated cell adhesion. PMID- 15886481 TI - Co-transplantation of schwann cells promotes the survival and differentiation of neural stem cells transplanted into the injured spinal cord. AB - The present study investigates whether Schwann cells (SCs) could promote the survival and differentiation of neural stem cells in the injured spinal cord. Neural stem cells were dissociated and cloned from the hippocampal tissue of newborn rats. SCs were also dissociated and purified simultaneously from the sciatic nerves of 4-day-old rats. The results showed that the number of surviving neural stem cells and differentiated neuron-like cells was significantly increased in the co-grafted (SCs and neural stem cells) group compared with the control group (neural stem cells only). Neuron-like cells that developed axon like processes were observed more commonly in the co-grafted group. These results demonstrate that SCs can promote the survival and differentiation of transplanted neural stem cells in the injured spinal cord. PMID- 15886482 TI - Plp gene regulation in the developing murine optic nerve: correlation with oligodendroglial process alignment along the axons. AB - The factors regulating the expression and splicing of the major myelin gene, proteolipid protein (Plp), are unclear. The gene encodes two splice variants, Plp and Dm20. During active myelination, transcription of the Plp gene is markedly upregulated and the splice variant ratio becomes Plp-mRNA dominant. We hypothesised that these aspects of Plp gene regulation are linked to overt axonal contact. Using the developing optic nerve of mice, we demonstrate that alignment of oligodendroglial processes with the axon correlates with both the expression of Plp-mRNA and the transcriptional upregulation of the gene. We test the above hypothesis more extensively in a subsequent study. PMID- 15886483 TI - Inhibition of glial maturation by bone morphogenetic protein 2 in a neural crest derived cell line. AB - Bone morphogenetic proteins (BMPs) regulate developmental decisions in many neural and nonneural lineages. BMPs influence both CNS neuronal and glial development and promote neuronal differentiation in neural crest derivatives. We investigated the actions of BMP2 on glial differentiation in the peripheral nervous system using NCM1 cells, a neural crest-derived cell line with the properties of peripheral glial precursor cells. BMP2 prevented the acquisition of a mature Schwann cell-like morphology, blocking the expression of mature genes and maintaining expression of several early glial markers. We provide evidence that BMP2 activates the GFAP promoter and define signaling pathways underlying this regulation. Our results demonstrate a novel role for BMPs as inhibitors of glial differentiation in the peripheral nervous system and suggest that BMPs may regulate the developmental timing of glial maturation. PMID- 15886484 TI - Mesencephalic trigeminal nucleus development is dependent on Krox-20 expression. AB - Krox-20, a C2H2-type zinc-finger transcription factor, plays an important role in rhombomere development. This study reveals that the Krox-20 null mutation impacts the development of mesencephalic trigeminal (Me5) neurons, a cell group traditionally thought to emerge from the mesencephalon. Based on cell counting studies, we show that Krox-20 null mutants have twice as many Me5 neurons relative to wildtypes at E15, but by birth have half the number of Me5 cells as wildtypes. TUNEL studies reveal a period of increased apoptosis from E17-P0 in mutants. The mutation does not result in differences in Me5 cell size, morphology, gene expression or peripheral projection patterns between genotypes, as demonstrated by retrograde tracing and Brn3a immunohistochemistry. The data suggest that Krox-20 regulates the period and extent of Me5 apoptosis, impacting the final number of Me5 neurons. The loss of Me5 in Krox-20-/- mice may highlight species-specific differences in the origin of these cells. PMID- 15886485 TI - Pediatric cortical dysplasia: correlations between neuroimaging, electrophysiology and location of cytomegalic neurons and balloon cells and glutamate/GABA synaptic circuits. AB - Seizures in cortical dysplasia (CD) could be from cytomegalic neurons and balloon cells acting as epileptic 'pacemakers', or abnormal neurotransmission. This study examined these hypotheses using in vitro electrophysiological techniques to determine intrinsic membrane properties and spontaneous glutamatergic and GABAergic synaptic activity for normal-pyramidal neurons, cytomegalic neurons and balloon cells from 67 neocortical sites originating from 43 CD patients (ages 0.2 14 years). Magnetic resonance imaging (MRI), (18)fluoro-2-deoxyglucose positron emission tomography (FDG-PET) and electrocorticography graded cortical sample sites from least to worst CD abnormality. Results found that cytomegalic neurons and balloon cells were observed more frequently in areas of severe CD compared with mild or normal CD regions as assessed by FDG-PET/MRI. Cytomegalic neurons (but not balloon cells) correlated with the worst electrocorticography scores. Electrophysiological recordings demonstrated that cytomegalic and normal pyramidal neurons displayed similar firing properties without intrinsic bursting. By contrast, balloon cells were electrically silent. Normal-pyramidal and cytomegalic neurons displayed decreased spontaneous glutamatergic synaptic activity in areas of severe FDG-PET/MRI abnormalities compared with normal regions, while GABAergic activity was unaltered. In CD, these findings indicate that cytomegalic neurons (but not balloon cells) might contribute to epileptogenesis, but are not likely to be 'pacemaker' cells capable of spontaneous paroxysmal depolarizations. Furthermore, there was more GABA relative to glutamate synaptic neurotransmission in areas of severe CD. Thus, in CD tissue alternate mechanisms of epileptogenesis should be considered, and we suggest that GABAergic synaptic circuits interacting with cytomegalic and normal-pyramidal neurons with immature receptor properties might contribute to seizure generation. PMID- 15886486 TI - Gestational diabetes mellitus in women in the fourth decade--is treatment worthwhile? AB - AIM: To examine the influence of diet-treated gestational diabetes mellitus on the obstetric performance of mothers aged 40 and above. METHOD: We reviewed the delivery records of 205 mothers aged 40 and above who delivered over a 3-year period. A 75-gram oral glucose tolerance test was performed in all cases and 64 (31.2%) (18 primiparas and 46 multiparas) had gestational diabetes mellitus. This affected group of patients was compared with a group of age- and parity-matched controls to determine the impact of gestational diabetes mellitus on the obstetric outcome. RESULTS: There was no difference in the maternal anthropometric parameters, antenatal complications, or labor performance. While no statistically significant difference was found in the infant anthropometric parameters, the study group had a lower incidence (p = 0.043) of large-for gestational age infants. CONCLUSION: Our findings suggested the adverse effects of gestational diabetes mellitus on pregnancy outcome were confounded to a large extent by other factors such as age, parity, and obesity. Once compared with matched controls, gestational diabetes mellitus that can be successfully treated with diet therapy probably had minimal adverse effect on the obstetric outcome. Furthermore, diet treatment can probably reverse the effect of advanced maternal age on infant size in these women. PMID- 15886487 TI - Effects of vitamin K2 administration on calcium balance and bone mass in young rats fed normal or low calcium diet. AB - OBJECTIVE: The purpose of this study was to examine the effects of vitamin K2 administration on calcium balance and bone mass in young rats fed a normal or low calcium diet. METHODS: Forty female Sprague-Dawley rats, 6 weeks of age, were randomized by stratified weight method into four groups with 10 rats in each group: 0.5% (normal) calcium diet, 0.1% (low) calcium diet, 0.5% calcium diet + vitamin K2 (menatetrenone, 30 mg/100 g chow diet), and 0.1% calcium diet + vitamin K2. After 10 weeks of feeding, serum calcium and calciotropic hormone levels were measured, and intestinal calcium absorption and renal calcium reabsorption were evaluated. Bone histomorphometric analyses were performed on cortical bone of the tibial shaft and cancellous bone of the proximal tibia. RESULTS: Feeding a low calcium diet induced hypocalcemia, increased serum parathyroid hormone (PTH) and 1,25-dihydroxyvitamin D [1,25(OH)2D] levels with decreased serum 25-hydrovyvitamin D [25(OH)D] level, stimulated intestinal calcium absorption and renal calcium reabsorption, and reduced cortical bone mass as a result of decreased periosteal bone gain and enlarged marrow cavity, but did not significantly influence cancellous bone mass. Vitamin K2 administration in rats fed a low calcium diet stimulated renal calcium reabsorption, retarded the abnormal elevation of serum PTH level, increased cancellous bone mass, and retarded cortical bone loss, while vitamin K2 administration in rats fed a normal calcium diet stimulated intestinal calcium absorption by increasing serum 1,25(OH)2D level, and increased cortical bone mass. CONCLUSION: This study clearly shows the differential response of calcium balance and bone mass to vitamin K2 administration in rats fed a normal or low calcium diet. PMID- 15886488 TI - Pharmacodynamic considerations with recombinant human insulin-like growth factor I in children. AB - AIM: To report effects of weight-based recombinant human insulin-like growth factor-I (rhIGF-I) on IGF axis parameters in children with hyperinsulinism. METHODS: Open label trial with subcutaneous rhIGF-I (40 microg/kg/dose). Patients studied were children (1 month to 11 years) with diffuse hyperinsulinism (n = 7). Serial serum IGF and insulin-like growth factor binding protein (IGFBP) concentrations were measured by RIA and analyzed by linear Pearson regression. RESULTS: Following the initial rhIGF-I dose, total insulin-like growth factor-I (IGF-I) rose by 56% at 30 min (p < 0.01) and 85% at 120 min (p < 0.02). Serum IGF II, IGFBP-2, and IGFBP-3 levels did not change. Peak serum IGF-I levels within 12 h of the initial rhIGF-I dose were 167-700 mg/ml. The variable peak IGF-I response is attributable in part to IGFBP-3 differences across this pediatric age range. Models of rhIGF-I dosing based upon body surface area (BSA) or initial IGFBP-3 resulted in predictable peak serum IGF-I levels (r = 0.78; p < 0.03). Recalculating rhIGF-I dosing based upon the BSA . IGFBP-3 product correlated closely with peak IGF-I level (r = 0.85; p < 0.007). CONCLUSIONS: Weight-based IGF-I dosing in this cohort resulted in variable IGF-I levels. Considering BSA and serum IGFBP-3 concentration in children is appropriate for subcutaneous IGF-I administration. A combination of these values may yield predictable individualization of rhIGF-I dosing. PMID- 15886489 TI - Impact of tuberculosis on serum leptin levels in patients with HIV infection. AB - AIM: Tuberculosis (TB) and human immunodeficiency virus (HIV) are classical wasting diseases accompanied by immunosuppression. As leptin is involved in the weight regulation and cellular immunity, we investigated the role of leptin levels in the co-infection of HIV and TB (HIV-TB). METHODS: The study group consists of the patients with asymptomatic HIV infection (n = 20), patients with HIV-TB co-infection (n = 20) and healthy control subjects (n = 20). Serum leptin levels and the concentrations of IFN-gamma, TNF-alpha, IL-12 and IL-4 cytokines were measured by ELISA before the start of the treatment. CD4+ T-cell counts were determined in patients with HIV and HIV-TB by flow cytometry. Body mass index (BMI) of the study subjects was calculated. RESULTS: Serum leptin levels and BMI were significantly lower in the patients with HIV-TB than control and HIV subjects. Multivariate regression analysis showed that serum leptin concentration was significantly dependent on BMI and sex but not on age and the disease groups. The leptin levels did not correlate either with CD4+ T-cell counts or with any of the serum cytokines in HIV and HIV-TB patients. CONCLUSION: Thus our finding suggests that the leptin concentrations were strongly associated with BMI and gender but not with the disease state or with the circulating cytokine levels. PMID- 15886490 TI - Lipopolysaccharide-induced cytokine and chemokine expression in human carotid lesions. AB - The release of cytokines and chemokines from activated immune-competent cells plays a crucial role in determining the pathology of the atherogenic progress. We investigated the effect of bacterial lipopolysaccharide (LPS) on cytokine/chemokine expression in carotid lesions and normal renal arteries. The lesions or renal arteries were incubated for 6 h at 37 degrees C in serum-free media treated with or without LPS. After LPS treatment, increased protein levels of IL-1beta, IL-6, IL-8, IL-10, TNF-alpha and MCP-1 were observed in the culture medium from the lesions measured with cytometric bead array. We were able to detect the induction of IL-1beta, IL-6, IL-8, IL-10, TNF-alpha and MCP-1 mRNA in the lesions after stimulation with LPS using real-time PCR. In renal arteries, LPS also induces mRNA expression of all chemokines and cytokines investigated with the exception of IL-6. However, LPS induces significantly higher levels of TNF-alpha, IL-1beta and IL-10 mRNA in lesions compared to renal arteries. The results suggest that infectious agents are capable of enhancing the production of cytokines/chemokines in an already ongoing inflammatory process such as in atherosclerosis, and that low levels of circulating LPS may affect the levels of pro-inflammatory cytokines much more in atherosclerotic vessels than in normal vessels and may contribute to the development of the atherosclerotic lesion. PMID- 15886491 TI - Reciprocal association of plasma adiponectin and serum C-reactive protein concentration in haemodialysis patients with end-stage kidney disease--a follow up study. AB - AIM: Malnutrition and inflammation are involved in the pathogenesis of atherosclerosis and increased risk of cardiovascular diseases in end-stage renal disease. The aim of this study was to assess the relationship between concentrations of plasma adiponectin, serum C-reactive protein (CRP), carotid intima-media thickness (IMT) and duration of haemodialysis (HD) treatment in prevalent HD patients. METHODS: Plasma adiponectin and serum CRP concentrations were estimated in 80 HD patients and 22 healthy controls. Carotid IMT was measured by ultrasound technique. HD patients were followed up for 23 +/- 16 months. During this period, 24 of them died. RESULTS: In HD patients, plasma adiponectin concentration was over 3 times higher than in controls (29.0 +/- 2.1 vs. 8.7 +/- 2.6 microg/ml; p < 0.001). HD patients with serum CRP concentrations > or = 5 mg/l were characterized by a lower plasma adiponectin concentration than HD patients with the CRP < 5 mg/l (23.9 +/- 3.5 vs. 33.0 +/- 3.1 microg/ml; p = 0.03). Plasma adiponectin and serum CRP concentrations were inversely related in HD (tau = -0.181; p = 0.02). No relationship between adiponectinaemia and IMT was observed. Survival (Kaplan-Meier analysis) within the lowest plasma adiponectin tertile tended (p = 0.06) to be the worse. CONCLUSIONS: (1) Inflammatory processes are associated with an inadequate low plasma adiponetin concentration in HD patients, and (2) a lower plasma adiponectin concentration seems to be a new predictor of poor outcome in HD patients. PMID- 15886492 TI - Fabry disease in patients with end-stage renal failure: the potential benefits of screening. AB - BACKGROUND/AIMS: Alpha-galactosidase A (alpha-GLA) deficiency (Fabry disease) is an X-linked lysosomal storage disorder. The associated visceral complications are progressive and multiorgan; renal involvement is common, usually leading to end stage renal failure (ESRF). The reported benefits of specific enzyme replacement therapy (ERT) indicate the importance of screening for Fabry disease in high-risk populations, as this approach should make it possible to identify other family members with little or no clinical features of the disease, and for them to be considered for early preventive treatment. METHODS: We screened for Fabry disease in 106 patients on hemodialysis in our hospital-based hemodialysis unit. We did this by measuring alpha-GLA enzyme activity in blood leukocytes taken from each patient and we then carried out gene analysis when indicated. RESULTS: We were able to discover 1 patient with low residual alpha-GLA activity (a prevalence of 0.94%). Alpha-GLA gene analysis identified a point mutation within the coding region producing a N215S amino acid substitution in the protein. Among the relatives of this index case, molecular testing found 7 family members with the same N215S alpha-GLA mutation. Of these, 3 had reduced alpha-GLA activity and clinical features of Fabry disease, and for which ERT was subsequently given. CONCLUSION: Screening for Fabry disease is simple and although the yield is small, it is potentially significant and of possible benefit to the relatives of affected cases in this 'at-risk' ESRF population, many of who do not have a clear renal diagnosis. PMID- 15886493 TI - Bedside estimation of the glomerular filtration rate in hospitalized elderly patients. AB - AIMS: To evaluate the quality of bedside estimation of glomerular filtration rate (GFR) in hospitalized elderly patients. METHODS: We evaluated common estimators of GFR in 29 women and 32 men aged 60 and older hospitalized in a geriatric ward: creatinine clearance (CCR), the Cockcroft-Gault formula (CG), the modification of diet in renal disease formula (MDRD), Baracskay formula (BAR), and a newly developed formula derived recently by us (GCM). Inulin clearance (CINU) was used to assess GFR. Exclusion criteria were mental illness and urinary incontinence. RESULTS: According to Bland and Altman accuracy and precision of all estimators were low and there was an underestimation of actual GFR: CCR 38.9 ml/min; CG 39.7 ml/min; MDRD 19.8 ml/min; BAR 34.0 ml/min, and GCM: 24.7 ml/min. The accuracy and precision of all methods were even lower in patients with a GFR of >90 ml/min and in patients with diabetes. In receiver-operating characteristics (ROC analysis) all formulas were superior to serum creatinine and overall MDRD disclosed the best results in detecting both a GFR of <90 ml/min and <60 ml/min. CONCLUSIONS: In general, estimation errors are large in an acute care setting. However, formula estimation is clearly superior to serum creatinine and CCR. MDRD gave the best results but may be replaced by the more simple CG and GCM formulas, whereas BAR was inferior. PMID- 15886494 TI - Treatment with cyclophosphamide in elderly-onset nephrotic syndrome. AB - BACKGROUND: The best treatment of elderly-onset nephrotic syndrome has not been well defined. The use of corticosteroids or combination immunosuppressants may be associated with a significant incidence of side effects in the elderly. There is little data on the use of cyclophosphamide alone. METHODS: We retrospectively reviewed 30 patients with idiopathic elderly-onset nephrotic syndrome treated with cyclophosphamide. RESULTS: Male to female ratio was 2:1, mean age at diagnosis was 72.7 +/- 5.9 years and average length of follow-up was 41.4 +/- 21.3 months. Significant co-morbidities, including hypertension, were present in 57%. A raised serum creatinine level was found in 57%. Biopsy revealed 15 membranous nephropathy, 4 mesangial proliferative Gn, 5 IgA nephropathy, 3 minimal change nephropathy, 2 focal segmental glomerulosclerosis and 1 C1q nephropathy. Cyclophosphamide was given for 32.0 +/- 16.2 weeks with an averaged cumulative dose per patient 177 +/- 84 mg/kg BW. Remission (complete or partial) was attained by 40, 63, 80 and 87% of patients within 12, 24, 36 and 48 weeks of treatment, respectively. Eighteen patients attained complete remission and 9 partial remission after treatment. The mean interval to attain first remission (complete or partial) was 18.9 +/- 14.6 weeks. This was not affected by age (p = NS) or initial albumin level (p = NS). At the time of last follow-up, all but 2 patients were in complete or partial remission with raised serum creatinine levels in 40%. CONCLUSIONS: Cyclophosphamide was effective and well tolerated in the treatment of elderly-onset nephrotic syndrome, with sustained remission and preserved renal function. PMID- 15886495 TI - Risk factors for early failure of native arteriovenous fistulas. AB - INTRODUCTION: Current guidelines recommend native arteriovenous fistulas (AVF) as the vascular access of choice for hemodialysis on account of the lower incidence of complications. However, this kind of vascular access has a high rate of early failure (early thrombosis or non-maturation). Our aim was to examine whether clear risk factors for early AVF failure could be identified in our patients. SUBJECTS AND METHODS: Data of all patients who underwent creation of an AVF at the Geneva University Hospital from January 1998 to December 2002 were reviewed. Early failure was defined as a non-functioning fistula (thrombosis or absence of fistula maturation). RESULTS: 119 patients underwent the creation of 148 native AVF, 88 (59.5%) in the forearm and 60 (40.5%) in the upper arm. 48 (32.4%) fistulae were created in diabetic patients. In a multiple logistic regression analysis, significant predictive factors of early failure were a distal location (adjusted odds ratio (aOR) = 8.21, 95% CI = 2.63-25.63, p < 0.001), female gender (aOR = 4.04, 95% CI = 1.44-11.30, p = 0.008), level of surgical expertise (aOR = 3.97, 95% CI = 1.39-11.32, p = 0.010) and diabetes mellitus (aOR = 3.19, 95% CI = 1.17-8.71, p = 0.024). CONCLUSION: Early failure of AVF occurs mainly in forearm sites among women and diabetic patients. Surgical expertise has also a significant influence. These results suggest that selection of a distal site for a native AVF has to be rigorously made for women and diabetic patients and that surgeon's dedication is of primary importance to avoid early AVF failure occurrence. PMID- 15886496 TI - Creating a fistula that works: is it an art or is it science? PMID- 15886497 TI - Secretion of parathyroid hormone oscillates depending on the change in serum ionized calcium during hemodialysis and may affect bone metabolism. AB - BACKGROUND: The current study was undertaken to clarify the dynamic response of parathyroid hormone (PTH) during hemodialysis and to determine whether or not such dynamic change of PTH affects bone turnover. METHODS: Serum ionized calcium (iCa) and intact PTH (iPTH) were measured in 66 dialysis patients before (basal) and after each hemodialysis. The changes of iCa (DeltaiCa) and iPTH (DeltaiPTH) were defined as [postdialysis iCa -basal iCa] mM, and [(postdialysis iPTH - basal iPTH)/basal iPTH] x 100%, respectively. We also investigated the data of the patients divided into four groups based on their basal iPTH levels; group 1 (iPTH < 60 pg/ml, n = 17), 2 (> or = 60 to < 150, n = 20), 3 (> or = 150 to < 300, n = 15), and 4 (> or = 300, n = 14). RESULTS: While iCa and iPTH changed variably during each hemodialysis procedure, there was a highly significant inverse correlation between DeltaiCa and DeltaiPTH (r = -0.761, p < 0.0001). Regression coefficients between DeltaiPTH and DeltaiCa were -519.1, -311.2, -268.1, and 194.6%/mM in groups 1-4, respectively. The difference in the regression coefficient was statistically significant between group 1 and 2 (F = 3.69, p < 0.05, ANCOVA), group 1 and 3 (F = 5.599, p < 0.05), and group 1 and 4 (F = 10.853, p < 0.005). This suggested that patients with higher basal iPTH responded poorly to modulate serum PTH levels by sensing the change of iCa. However, after an intensified oral calcitriol therapy to reduce iPTH, the PTH response in group 4 patients was restored to levels comparable with those observed in patients having lower basal iPTH. We also demonstrate that the DeltaiPTH of patients of group 1 but not of other groups was significantly correlated with serum markers for bone metabolism; osteocalcin (r = 0.535, p < 0.05) and collagen type I C terminal telopeptide (r = -0.575, p < 0.05). CONCLUSIONS: Our findings suggest that secretion of PTH is dynamically regulated by DeltaiCa during hemodialysis and such oscillated PTH secretion may affect bone metabolism in a subset of dialysis patients. PMID- 15886498 TI - Effects of simvastatin on oxidative stress in streptozotocin-induced diabetic rats: a role for glomeruli protection. AB - AIMS: To study the effects of simvastatin on oxidative stress in rats with early stage diabetic nephropathy. METHODS: 60 male Sprague-Dawley rats were divided into three groups: control group (CN), streptozotocin (STZ)-induced diabetic rats group (DM) and STZ-induced diabetic rats group treated with simvastatin (DM+S). The following parameters were measured at weeks 6 and 12 in similar rats chosen randomly from each group: body and kidney weight, 24-hour urinary albumin excretion (UAE), biochemical indexes including blood glucose (GLU), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TG), serum creatinine (SCr), antioxidant enzymes including superoxide dismutase (SOD), glutathione S-transferase (GST), catalase (CAT) in plasma, lipid peroxidation production as malondialdehyde in plasma (MDAp) and erythrocytes (MDAe), morphology parameters such as glomerular volume (GV) and mesangial area/total glomerular area (M/T). RESULTS: At weeks 6 and 12, GLU and kidney weight to body weight ratio were notably increased in both of the diabetic groups compared with those in the CN group without significant differences between the two diabetic groups. There were no significant differences of SCr, LDL, HDL and TG among all groups within all the experimental time. MDAp and MDAe were significantly increased in both of the diabetic groups, especially at week 12, while SOD, GST and CAT were significantly decreased compared with those in the CN group. At week 12, GV, M/T and UAE were also increased in the two diabetic groups. However, in the DM+S group, changes of lipid peroxidation production, antioxidant enzymes, UAE and GV were less pronounced than those in the DM group. Pearson's correlation analysis and regression analysis shown that MDAp was increased while SOD, GST and CAT in plasma were decreased with elevation of UAE, GV and M/T. CONCLUSION: Increased lipid peroxidation and decreased antioxidant enzymes in plasma may play a role in the progression of diabetic nephropathy. Simvastatin may ameliorate these changes to protect kidney from oxidative lesion in diabetes even in the absence of lipid abnormalities. PMID- 15886499 TI - The N- and L-type calcium channel blocker cilnidipine suppresses renal injury in dahl rats fed a high-sucrose diet, an experimental model of metabolic syndrome. AB - BACKGROUND/AIMS: The L/N-type calcium channel blocker (CCB) cilnidipine has been demonstrated to suppress progressive renal disease in a variety of experimental models, but the characteristic effects of N-type calcium channel blocking action on renal injury have not been examined in detail. Therefore, we investigated the beneficial effects of cilnidipine on renal injury in Dahl salt-sensitive (Dahl S) rats fed a high-sucrose diet (HSD), which mimics metabolic syndrome, and compared them with the effects of an L-type CCB, amlodipine. METHODS: Male Dahl S rats were divided into groups with similar blood pressure at 8 weeks of age and fed an HSD. They received vehicle, cilnidipine or amlodipine for 27 weeks. At 35 weeks of age, urine and blood samples were collected for physiological analysis, and the kidneys were removed for histopathological evaluation. RESULTS: Cilnidipine reduced albuminuria, glomerular hypertrophy, glomerular expression of ICAM-1, ED 1-positive cell infiltration and interstitial fibrosis compared with vehicle treated rats. In contrast, amlodipine had no effect on these parameters. Urinary norepinephrine excretion, renal expression of renin mRNA and renal tissue levels of angiotensin II were increased only in the amlodipine-treated group. CONCLUSION: Cilnidipine provided superior protection against renal damage compared with amlodipine in Dahl S rats given an HSD. The different effects between these two drugs may be partly explained by their different actions on the renal sympathetic nerve activity and the renin-angiotensin system through the N type calcium channel blocking action. PMID- 15886500 TI - Papillary microcarcinomas of the thyroid with preoperatively detectable lymph node metastasis show significantly higher aggressive characteristics on immunohistochemical examination. AB - OBJECTIVES: We recently demonstrated that papillary microcarcinomas with preoperatively detectable node metastasis in the lateral compartment on ultrasonography (clinically apparent metastasis) show worse postoperative relapse free survival than those with no metastasis or metastasis that could not be detected preoperatively, but was confirmed by pathological examination after surgery (occult metastasis). In this study, we investigated difference in the aggressive characteristics of microcarcinoma of this type from various perspectives. MATERIALS AND METHODS: We immunohistochemically examined the expression of cell proliferating markers, Ki-67, cyclin D1, p27, and retinoblastoma gene product (pRb), apoptotic markers, single-strand DNA (ssDNA), and metastatic suppressor, kangai-1 (KAI-1) for 19 microcarcinoma patients with clinically apparent metastasis, 14 patents with occult metastasis, and 22 patients without metastasis. RESULTS: Cases of clinically apparent metastasis showed increased cyclin D1 expression together with decreased p27 expression and higher levels of pRb and Ki-67 expression. Furthermore, ssDNA expression was higher and bcl-2 expression was lower in these cases, while KAI-1 expression was significantly reduced. There was no significant difference in the expression of these proteins between cases demonstrating no and occult metastases. CONCLUSION: These findings suggest that cases of clinically apparent metastasis show significantly higher growth based on cell proliferating activity, apoptosis, and expression of metastatic suppressor than those demonstrating no or occult metastases. PMID- 15886501 TI - Syndecan-1 expression--a novel prognostic marker in pancreatic cancer. AB - OBJECTIVE: Syndecan-1 is a transmembrane receptor that participates in cell-cell and cell-matrix interactions, cell proliferation and cell migration. Expression of syndecan-1 is downregulated in many cancers, but in pancreatic ductal adenocarcinoma it is upregulated. METHOD: We studied the immunohistochemical expression of syndecan-1 in 144 pancreatic adenocarcinomas and evaluated the prognostic value of syndecan-1 expression. Formalin-fixed, paraffin-embedded specimens were stained with mouse monoclonal antibody B-B4 against human syndecan 1. The epithelial and stromal staining was separately evaluated and compared with patient survival, clinical stage and histological grade. RESULT: Epithelial immunoreactivity was observed in most of the pancreatic carcinoma samples: in 70 (49%) of the samples the epithelial staining was weak, in 48 (33%) moderate, in 18 (12%) strong and in only 8 (6%) of the samples the epithelial staining was negative. Stromal staining was weak in 24 (17%), moderate in 31 (22%), strong in 11 (8%) and negative in 78 (54%) of the pancreatic carcinoma samples. Lack of stromal expression predicted a better prognosis (p = 0.002; HR 1.7) and it was independent of stage (p = 0.01; HR = 1.5) and grade (p = 0.0004; HR 2.1) in multivariate analysis. Epithelial expression predicted better prognosis for patients that underwent surgery for cure (n = 94, p = 0.03). CONCLUSION: Stromal syndecan-1 expression is an independent prognostic marker in pancreatic cancer, whereas epithelial syndecan-1 expression predicts better prognosis only in resectable disease. PMID- 15886502 TI - Economic burdens and quality of life of family caregivers of cancer patients. AB - OBJECTIVES: We conducted this study to identify factors influencing the burdens cancer brings to a patient's family and to evaluate the association between the burdens and the caregiver's quality of life (QOL). METHODS: Participants were drawn from the primary family caregivers of cancer patients at 6 university hospitals and the National Cancer Center in Korea. Of the 738 eligible caregivers, 704 (95.4%) completed the questionnaire packets (Family Impact Questions and Caregiver's QOL-Cancer). RESULTS: Caregivers, who were poor (OR, 2.11; 95% CI, 1.44-3.10), whose health status was poor (OR, 1.87; 95% CI, 1.29 2.70), who were married (OR, 1.75; 95% CI, 1.12-2.72), who provided care for a long time (OR, 2.29; 95% CI, 1.59-3.28), who cared for patients with poor performance status (OR, 1.35; 95% CI, 1.00-1.82), and who paid high medical expenses (OR, 1.70; 95% CI, 1.21-2.40), were more likely to lose their family savings. In multiple regression analysis, most burden variables--including requiring caregiving assistance, major life change, inability to function normally, loss of savings, loss of income, and altered educational plans--were associated with caregiver QOL. Loss of family income, which was related to economic issues, was most strongly associated with it (16.0%). CONCLUSIONS: Our study suggests that to improve caregiver QOL, we should give priority to decreasing the economic burden that cancer places on patient's family. PMID- 15886503 TI - Is there an indication for high-dose chemotherapy in the treatment of bone and soft-tissue sarcoma? AB - Sarcomas represent a rare and heterogeneous disease, and the prognosis of patients with unresectable or advanced metastatic bone and especially soft-tissue sarcomas remains poor, with a disease-free survival of less than 10% at 5 years. Only few chemotherapeutic agents have been identified to be active, with reported response rates for doxorubicin, epirubicin and ifosfamide above 20%. Although combination therapy with different chemotherapeutic substances results in higher response rates, superiority against single-agent chemotherapy in terms of survival has not been demonstrated yet. Regarding anthracyclines, and ifosfamide in particular, a dose-response relationship has been shown, and high-dose chemotherapy with stem cell support has also been evaluated by several investigators. However, all studies comprised small patient cohorts and included very heterogeneous histological subtypes of soft-tissue sarcoma. Moreover, randomized trials are missing. Nevertheless, higher doses of chemotherapy result in higher response rates possibly correlating with longer survival. Finally, well designed randomized trials should be performed, preferably in younger patients and in the context of an interdisciplinary treatment approach. In this review, we provide an overview of the literature concerning high-dose chemotherapy with hematopoietic stem cell support in the treatment of bone and soft-tissue sarcomas. On the basis of our own data, we would like to emphasize the importance of high-dose chemotherapy in the treatment of sarcomas. PMID- 15886504 TI - Binding properties of dipropyltryptamine at the human 5-HT1a receptor. AB - Dipropyltryptamine (DPT) is a synthetic indolealkylamine first characterized in the 1960s. Largely forgotten since the discovery of multiple serotonin receptor subtypes, some of the properties of DPT at the cloned human 5-HT1a receptor are described here. When [3H]8-OH-DPAT is bound to the receptor, DPT inhibits the interaction with an IC50 of 0.1 micromol/l. This interaction is shown to be competitive when double-reciprocal plots of the DPT/agonist interaction are analyzed. DPT's effects in the signal transduction system are complex. While DPT alone (0.1-1,000 micromol/l) activates Gi when both cAMP and gamma-S-GTP incorporation are measured, in the presence of 5-HT (0.1-10 micromol/l), DPT blocks the agonist effect. In combination, the findings suggest that DPT is a moderate affinity partial agonist at the human 5-HT1a receptor. These results provide evidence that DPT has potential as a versatile experimental tool at 5 HT1a receptors. PMID- 15886505 TI - Synergic antinociceptive interaction between tramadol and gabapentin after local, spinal and systemic administration. AB - The possible interaction between tramadol and gabapentin on formalin-induced nociception in the rat was assessed. Tramadol, gabapentin or a fixed-dose ratio combination of gabapentin and tramadol were administered peripherally, spinally and orally to rats, and the antinociceptive effect was determined in the 1% formalin test. Isobolographic analyses were used to define the nature of the interactions between drugs. Tramadol, gabapentin and tramadol-gabapentin combinations produced a dose-dependent antinociceptive effect when administered locally, spinally or orally. ED30 values were estimated for the individual drugs and isobolograms were constructed. Theoretical ED30 values for the combination estimated from the isobolograms were 126.8 +/- 11.1 microg/paw, 23.1 +/- 2.6 microg/rat, and 2.23 +/- 0.32 mg/kg for the local, intrathecal and oral routes, respectively. These values were significantly higher than the actually observed ED30 values which were 13.3 +/- 2.1 microg/paw, 8.1 +/- 0.6 microg/rat and 0.71 +/- 0.10 mg/kg, indicating a synergistic interaction. Although efficacy was not improved, local peripheral administration resulted in the highest increase in potency, being about tenfold. Spinal and systemic administration increased potency threefold. Data indicate that low doses of the tramadol-gabapentin combination can interact synergistically to reverse formalin-induced nociception and may represent a therapeutic advantage for clinical treatment of inflammatory pain. PMID- 15886506 TI - Intracranial meningiomas of childhood and adolescence. AB - Meningiomas are rare intracranial neoplasms in childhood and adolescence, representing 0.4-4.1% of the pediatric-age tumors and 1.5-1.8% of all intracranial meningiomas. The goal of this study was to determine epidemiology, clinical and radiological features, and long-term outcome of childhood and adolescence meningiomas. Patients operated for intracranial meningiomas of childhood and adolescence between 1983 and 2003 at Gazi University School of Medicine, Department of Neurosurgery, were evaluated retrospectively. This study presents 11 cases (6 male, 5 female), ranging in age from 14 months to 17 years. Age and sex distribution, presenting symptoms, neurological examination results, location of meningiomas, radiological and histopathological findings, and prognosis were reviewed. The results were compared with those reported in the existing literature. Atypical and malignant meningiomas seem to be more common in childhood and adolescence with respect to adult meningiomas. Tumor location, completeness of tumor removal, and pathological grade are the most important prognostic factors. PMID- 15886507 TI - The gravity-assisted Paedi-Gav valve in the treatment of pediatric hydrocephalus. AB - OBJECTIVE: A single-center, prospective, nonrandomized pilot study was performed to assess the Paedi-Gav gravity-assisted valve for the treatment of pediatric patients with hydrocephalus. METHODS: Participants were pediatric patients (age <16 years) who were candidates for a hydrocephalus shunt system that required a valve insertion at the time of enrollment. The primary outcome event was shunt malfunction; subclassified into shunt obstruction, shunt overdrainage, loculated ventricles, or infection. The shunt obstructions were further subclassified according to site. A total of 32 patients were enrolled onto the study, with 2 undergoing first shunt insertion after failed ventriculostomy and 30 undergoing shunt revisions. On average, the patients had had 3.3 shunt procedures prior to insertion of a Paedi-Gav valve. RESULTS: During a follow-up interval of minimum 52 weeks and a median of 24 months after the first implantation on-study, shunt revisions were required in 17 (53.1%) of the 32 patients. The 12-month shunt survival rate without revision of any component was 53%, with a median shunt survival time of 388 days. The most common reasons for shunt revision were shunt obstructions (12/17) and overdrainage (3/17). Shunt obstructions were caused by valve-related failures (9/12) and distal obstructions (3/12). CONCLUSION: Although the small number of patients enrolled in this study warrants cautious conclusions, the overall results are comparable to those reported for primary shunt insertions with conventional valves in pediatric patients with hydrocephalus. Although this study provides a rationale for examining the Paedi Gav gravity-assisted shunt valve in a larger prospective randomized controlled trial, the shunt failure pattern, with a rather high frequency of valve-related failures, may indicate potential for further improvements in the valve design and/or manufacturing. PMID- 15886508 TI - Persistent cognitive dysfunction secondary to cerebellar injury in patients treated for posterior fossa tumors in childhood. AB - Traditionally, the cerebral hemispheres have been regarded as the region of the brain responsible for cognitive functions, while the cerebellum has been considered to be primarily involved in motor functions. Recent studies focus also on the possible involvement of the cerebellum in neurocognitive functions. The aim of this study was to determine the neuropsychological profile of young adults treated for a posterior fossa tumor in childhood and look for possible support for the presence of the so-called 'cerebellar cognitive affective syndrome' in these patients. Two groups of young adults were studied. The astrocytoma group (n = 12) had been treated for a low-grade cerebellar astrocytoma with surgery alone (mean age at surgery was 8.6 years and mean age at neuropsychological testing was 23.5 years). The medulloblastoma group (n = 11) had been treated with surgery followed by radiotherapy and chemotherapy (mean age at surgery was 6.1 years and mean age at neuropsychological testing was 23.1 years). The neuropsychological test battery comprised measures of intelligence, motor function, attention, psychomotor speed, verbal memory and visual memory. The medulloblastoma group performed poorer than the astocytoma group on all neuropsychological measures except one. Nonetheless, the astrocytoma group also had impaired scores compared with standard norms on measures of motor speed, attention and executive function. No significant correlation between age at time of treatment and grade of neuropsychological impairment was found in the astrocytoma group, though there was a tendency that young age at time of treatment correlated with better outcome on IQ measures. In the medulloblastoma group, age was significantly correlated with outcome, for both IQ and degree of neuropsychological impairment. For this group, young age at time of treatment indicated a worse outcome. CONCLUSIONS: Persistent cognitive dysfunction was detected in patients treated for posterior fossa medulloblastoma and cerebellar astrocytoma. The astrocytoma group was treated with surgery alone, indicating that a cerebellar lesion can result in cognitive dysfunction. Thus, this study gives support to the existence of the cerebellar cognitive affective syndrome. Follow-up of all patients treated for posterior fossa tumor in childhood should include an extensive neuropsychological testing at regular intervals. This may be of benefit for school planning and later work planning. PMID- 15886509 TI - Benign intradural extramedullary masses in children of northern India. AB - Twenty-five children with benign intradural extramedullary tumors/lesions were retrospectively analyzed to see the frequency of different histotypes in this location and their variation from the Western world. The duration of symptoms, clinical profile and surgical outcome of these cases were evaluated. The age of children ranged from 1.5 to 18 years, with a mean age of 7.5 years. Thirteen amongst them were male. Follow-up ranged from 3 months to 5 years with a mean of 19 months. McCormick functional grading was used retrospectively to find out the functional outcome in these children. Sixty-eight percent of these cases presented with functional grade IV or V. The mean duration from the appearance of the first symptoms to admission to our hospital in these children was 13 months. Improvement in the functional grade following surgical excision was noted amongst all, except one, who presented with schwannoma, and did not improve from grade III even after a 14-month follow-up. Reoperation/two-staged operation was required in 3 children. The following conclusions were drawn from this study: (1) approximately half (48%) of intradural extramedullary benign tumors are nerve sheath tumors, i.e. neurofibromas, neurilemomas and schwannomas. Giant neurofibromas with giant extraforaminal extension may rarely manifest with a large extraspinal mass; (2) meningioma is an infrequent tumor in this location; (3) developmental cystic dysraphic lesions (neurenteric cyst, arachnoid cysts), unassociated with the stigmata of spinal dysraphism, are significantly more frequent (28%) than reported in the literature; (4) the possibility of infective lesions (granuloma, abscess) causing compressive features cannot be denied even in intradural extramedullary tumors; (5) long-standing lesions, with a long segment involvement, also have a good outcome following a good microsurgical excision, and (6) more than half of the children (68%) present with a severe neurological compromise, either because of ignorance and poor education or due to the scanty diagnostic facilities available at the peripheral hospitals of the developing milieu. PMID- 15886510 TI - Hydrocephalus as the initial presentation of a spinal cord astrocytoma associated with leptomeningeal spread. AB - A 3-year-old boy presented with headaches, vomiting, lethargy and papilledema. Communicating hydrocephalus along with transependymal fluid absorption and meningeal contrast enhancement was identified on CT. The enhancement was initially thought to be the result of a partially treated meningitis (child was previously on oral antibiotics for a presumed mycoplasma pneumonia). A right ventricular-peritoneal shunt was placed. CSF studies procured during the procedure were all normal. In contrast, CSF from a lumbar puncture contained a high protein, and cytology was highly suspicious for malignancy. Spine MRI showed diffuse leptomeningeal enhancement and a 1.5-cm intramedullary lesion at T12-L1 associated with minimal edema. The lesion was subtotally resected (70%) and diagnosed as an astrocytoma (mostly Kernohan grade 2 but with areas of grade 3). Chemotherapy was administered and follow-up spine MRI at 2 months did not reveal any residual tumor, however, the leptomeningeal enhancement persisted. Sixteen months later, at the completion of the chemotherapy and radiation therapy, the spine MRI remained unchanged. Neurological examination has always been normal. This case illustrates how a spinal cord astrocytoma can metastasize via spinocranial dispersion and present early with hydrocephalus rather than myelopathy. PMID- 15886511 TI - Human tail with noncontiguous intraspinal lipoma and spinal cord tethering: case report and embryologic discussion. AB - Children born with a tail-like appendage have a rare malformation that is frequently associated with abnormalities of the spine and spinal cord. A contiguous fibrolipoma is usually seen extending from the subcutaneous portion of the tail into the inferior spinal cord, resulting in tethered cord syndrome. We present the case of a child born with a tail and intraspinal lipoma that were not contiguous with each other, and were separated by an intact layer of lumbosacral fascia. The tail and lipoma were removed and the spinal cord untethered, and the child is neurologically normal 2 years after surgery. The absence of a contiguous lipoma from the tail to the spinal cord suggests that this condition may be principally caused by a disorder of secondary neurulation and/or regression of the normal embryonic tail bud. The embryology of the lower spine is reviewed and possible etiologies discussed. PMID- 15886512 TI - Penetrating brain injuries from a dog bite in an infant. AB - Brain lesions due to dog bites are not frequent and mainly concern infants in the first years of their life because they are short in height, the size of the infant's head is relatively large compared to the body and the skull bones are thin. We report the case of an infant with bilateral skin, skull and brain injuries secondary to a dog bite, and review the relative reports in the literature. We stress the need to consider the possibility of penetration into the intracranial compartment occurring in cases of dog bites of the scalp, because these wounds on the scalp and the skull may appear relatively limited, small in size and slight, in spite of associated potentially dangerous deeper lesions involving the intracranial structures that may be missed on the first observation. Infection is the main possible complication, and may be the cause of permanent and serious neurological deficits. Prompt diagnosis and therapy are mandatory to avoid complications and to achieve good clinical results. PMID- 15886513 TI - Osteochondroma of the sacrum with a correlative radiographic and histological evaluation. AB - Primary benign sacral tumors are rare representing only 1-4% of solitary exostoses and constituting 4% of all solitary spinal tumor. The tumor usually comes to attention in childhood or adolescence. The treatment of symptomatic lesions is resection. We present here a case of a sacral osteochondroma with characteristic histological and radiological features in an adolescent presenting with radicular pain. PMID- 15886514 TI - Spontaneous posterior fossa extradural hematoma--a rare complication following cardiac surgery. AB - Posterior fossa extradural hematoma (PFEDH) is an infrequent condition. Though numerous neuropathological sequelae of cardiopulmonary bypass have been described, to the best of the authors' knowledge, PFEDH after cardiac surgery has not been reported in the English literature. We report a 4-year-old boy with tricuspid atresia, who underwent cardiac surgery under cardiopulmonary bypass. On the 2nd postoperative day, his neurological condition deteriorated, and a CT scan of the head revealed a large PFEDH with mass effect and hydrocephalus. He underwent a midline suboccipital craniectomy and evacuation of PFEDH. There was diffuse bleeding from the dura; however, no specific site of bleeding was identified. The child started following commands on the 1st postoperative day, had an uneventful recovery and was discharged on the 10th postoperative day. On follow-up, 1 year after the surgery, he was doing well. PMID- 15886515 TI - Remote cerebellar hemorrhage after craniotomy. PMID- 15886516 TI - Subgaleal dermoid tumors at the anterior fontanelle. PMID- 15886517 TI - Occult tight filum terminale syndrome: results of surgical untethering. PMID- 15886519 TI - Analysis of the factors affecting the accuracy of detection for single base alterations by oligonucleotide microarray. AB - The oligonucleotide microarray, a high-throughput polymorphism detection technology, holds great promise for the characterization of complex genetic variance. To achieve greater sensitivity and specificity for it to be an effective platform technology we present results and discuss some of the factors influencing signal intensities and single-mismatch discrimination in array-based mutation/SNP detection. Probes with a series of concentrations were spotted onto the slide in order to find the optimal concentration with the identifiable satisfying signals and the stable ratios between matched and mismatched probes. It was found that under our experimental conditions, when the initial probe concentration is higher than the maximum immobilization capability of the slide (7.5 microM), the hybridization signal will be saturated and the ratio between matched and mismatched probes will be more stable than at a lower probe concentration. Considering the cost of probes and the systematic stability, a constant spotting concentration of 10 microM was selected. The stability of different types of mismatched oligo-DNA duplexes on the glass surface was also confirmed. The results show that the order of stability of mismatched oligo-DNA duplexes on a glass surface is in general agreement with previous reports conducted using liquid and polyacrylamide gel pads. This suggests that the influence of the mismatched base pair on the stability of the duplex in a solid hybridization system is similar to that in the solution hybridization environment. PMID- 15886520 TI - CD137 induces adhesion and cytokine production in human monocytic THP-1 cells. AB - CD137, which is expressed on activated T cells, plays a critical role in inflammatory responses. However, the exact role that CD137 plays in monocytes is not fully known. Here we studied the expression and function of CD137 in human monocytic THP-1 cells, which we found constitutively expresses CD137 at the mRNA and protein level. Cross-linking of CD137 increased the secretion of IL-8 and TNF alpha, promoted the expression of CD54 and CD11b, and increased adhesion to extracellular matrix (ECM) proteins. In particular CD137-induced adhesion of THP 1 cells was inhibited by an inhibitor of mitogen-activated protein kinase kinase (MEK), but not by a p38 kinase inhibitor. Taken together, these results show that the adhesion and cytokine production of THP-1 cells induced by CD137 occur via activation of MEK, which results in the activation of ERK-1/2 signaling pathways. Therefore, this study suggests that CD137 induces an activating and migrating signal during inflammatory processes. PMID- 15886521 TI - Obesity and genetic polymorphism of ERCC2 and ERCC4 as modifiers of risk of breast cancer. AB - To evaluate the relationship of genetic polymorphisms of ERCC2 and ERCC4 genes, both involved in nucleotide excision repair (NER), and the risk of breast cancer, a hospital-based case-control study was conducted in Korea. Histologically confirmed breast cancer cases (n = 574) and controls (n = 502) with no present or previous history of cancer were recruited from three teaching hospitals in Seoul during 1995-2001. Information on selected characteristics was collected by interviewed questionnaire. ERCC2 Asp(312)Asn (G>A) was genotyped by single-base extension assay and ERCC4 Ser(835)Ser (T>C) by dynamic allele-specific hybridization system. Although no significant association was observed between the genetic polymorphisms and the risk of breast cancer, women with both ERCC2 A allele- and ERCC4 C allele-containing genotypes showed a 2.6-fold risk (95% CI: 1.02-6.48) of breast cancer compared to women concurrently carrying the ERCC2 GG and ERCC4 TT genotypes. The breast cancer risk increased as the number of "at risk" genotypes increased with a borderline significance (P for trend = 0.07). Interactive effect was also observed between ERCC4 genotype and body mass idnex (BMI) for the breast cancer risk; the ERCC4 C allele containing genotypes posed a 1.7-fold (95% CI: 0.96-2.93) breast cancer risk in obese women (BMI>25 kg/m(2)) with a borderline significance. Our finding suggests that the combined effect of ERCC2 Asp(312)Asn and ERCC4 Ser(835)Ser genotypes might be associated with breast cancer risk in Korean women. PMID- 15886522 TI - Calcium sensing receptor forms complex with and is up-regulated by caveolin-1 in cultured human osteosarcoma (Saos-2) cells. AB - The calcium sensing receptor (CaSR) plays an important role for sensing local changes in the extracellular calcium concentration ([Ca(2+)](o)) in bone remodeling. Although the function of CaSR is known, the regulatory mechanism of CaSR remains controversial. We report here the regulatory effect of caveolin on CaSR function as a process of CaSR regulation by using the human osteosarcoma cell line (Saos-2). The intracellular calcium concentration ([Ca(2+)](i)) was increased by an increment of [Ca(2+)](o). This [Ca(2+)](i) increment was inhibited by the pretreatment with NPS 2390, an antagonist of CaSR. RT-PCR and Western blot analysis of Saos-2 cells revealed the presence of CaSR, caveolin (Cav)-1 and -2 in both mRNA and protein expressions, but there was no expression of Cav-3 mRNA and protein in the cells. In the isolated caveolae-rich membrane fraction from Saos-2 cells, the CaSR, Cav-1 and Cav-2 proteins were localized in same fractions (fraction number 4 and 5). The immuno-precipitation experiment using the respective antibodies showed complex formation between the CaSR and Cav 1, but no complex formation of CaSR and Cav-2. Confocal microscopy also supported the co-localization of CaSR and Cav-1 at the plasma membrane. Functionally, the [Ca(2+)](o)-induced [Ca(2+)](i) increment was attenuated by the introduction of Cav-1 antisense oligodeoxynucleotide (ODN). From these results, in Saos-2 cells, the function of CaSR might be regulated by binding with Cav-1. Considering the decrement of CaSR activity by antisense ODN, Cav-1 up-regulates the function of CaSR under normal physiological conditions, and it may play an important role in the diverse pathophysiological processes of bone remodeling or in the CaSR related disorders in the body. PMID- 15886523 TI - Identification and characterization of peroxisome proliferator response element in the mouse GLUT2 promoter. AB - In the present study, we show that the expression of type 2 glucose transporter isoform (GLUT2) could be regulated by PPAR-gamma in the liver. Rosiglitazone, PPAR-gamma agonist, activated the GLUT2 mRNA level in the primary cultured hepatocytes and Alexander cells, when these cells were transfected with PPAR gamma/RXR-alpha. We have localized the peroxisome proliferator response element in the mouse GLUT2 promoter by serial deletion studies and site-directed mutagenesis. Chromatin immunoprecipitation assay using ob/ob mice also showed that PPAR-gamma rather than PPAR-alpha binds to the -197/-184 region of GLUT2 promoter. Taken together, liver GLUT2 may be a direct target of PPAR-gamma ligand contributing to glucose transport into liver in a condition when PAPR-gamma expression is increased as in type 2 diabetes or in severe obesity. PMID- 15886524 TI - Antitumor activity of spinasterol isolated from Pueraria roots. AB - We purified phytoestrogens from Pueraria root (Pueraria mirifica from Thailand and Pueraria lobata from Korea), which is used as a rejuvenating folk medicine in Thailand and China. Dried, powdered plant material was extracted with 100% ethanol and further separated by concentration, filtration, and thin layer silica gel chromatography. Using the fractions obtained during separation, we first investigated their cytotoxicity in several cancer cell lines from various tissues. The ethanol-extracted components (PE1, PE4) had significant antiproliferative effects on breast cancer cell lines, including MCF-7, ZR-75-1, MDA-MB-231, SK-BR-3, and Hs578T. Second, we compared these results with the cytotoxic effects of known flavonoids, sterols, and coumarins from Pueraria root. The known compounds were not as effective, and occurred in a different polarity region on HPLC. Third, further separation resulted in the isolation of eight different components (Sub PE-A to -H). One of these, PE-D, affected the growth of some breast cancer cell lines (MCF-7, MDA- MB-231) in a dose- and time-dependent manner, as well as the growth of ovarian (2774) and cervical cancer cells (HeLa). Finally, a transfection assay showed that this component had an estrogenic effect similar to 17beta - estradiol, which activates both estrogen receptor alpha (ERalpha) and ERbeta. The NMR analysis determined that spinasterol (stigmasta-7, 22-dien-3beta-ol) is an active cytotoxic component of Pueraria root. PMID- 15886525 TI - In vivo characterization of the integrin beta3 as a receptor for Hantaan virus cellular entry. AB - Binding of viruses to cell surface molecules is an essential step in viral infection. In vitro studies suggested that the alpha(v)beta(3) integrin receptor is the epithelial cell receptor for Hantaan virus (HTNV). Whether beta(3) is in vivo the only or central cellular receptor for HTNV infection is not known. To investigate the role of beta(3) integrin for cellular entry of HTNV, we established an HTNV infection model in newborn murine pups. Infected pups died at an average age of 14.2 +/- 1.1 days with high levels of viral antigen detected in their brain, lung, and kidney. Pre-injection of blocking monoclonal antibodies (mAb) specific for either beta(3) or av prolonged survival significantly to a maximal average survival of 19.7 +/- 1.5 days (P <0.01) and 18.4 +/- 0.9 days (P < 0.01), respectively. XT-199, a chemical blocker of the alpha(v)beta(3) receptor also prolonged survival to 19.5 +/- 1.3 days (P < 0.01). In contrast to these receptor blockades, anti-HTNV antibody was not only able to prolong survival, but 20% of infected pups achieved long-term survival. An anti-murine beta(1) antibody comparatively prolonged survival (19.0 +/- 1.2 days), suggesting that HTNV infection is partly mediated through integrin beta(1) receptors as well as through beta(3) receptors in vivo. Our data demonstrate that the beta(3) receptor is important for HTNV infection in vivo, but also suggest that HTNV may utilize additional receptors beyond beta(3) for cellular entry within an organism. PMID- 15886526 TI - Common interleukin 10 polymorphism associated with decreased risk of tuberculosis. AB - Interleukin 10 (IL10) is a powerful TH2-cell cytokine that inhibits lymphocyte replication and secretion of inflammatory cytokines. The genetic associations of polymorphisms in IL10 with clinical manifestations of tuberculosis (TB) were examined in a large number of patients with clinical TB infection (n = 459) and normal controls (n = 871). One common promoter SNP (IL10 -592 A>C) was found to be significantly associated with decreased risk of TB manifestation. The frequency of the "C"-bearing genotype was higher in normal controls than in patients with clinical TB infection (P = 0.005, OR = 0.69). A summary of the genetic effect of IL10 -1082 A>G, the other nearby promoter SNP, in other ethnic groups is also presented. PMID- 15886527 TI - Cryptotanshinone but not tanshinone IIA inhibits angiogenesisin vitro. AB - In the course of screening of angiogenesis inhibitor from natural products, cryptotanshinone from Salvia miltiorrhiza was isolated as a potent small molecule inhibitor of angiogenesis. Cryptotanshinone inhibits bFGF-induced angiogenesis of BAECs at ten micromolar ranges in vitro without cytotoxicity. Tanshinone IIA, another tanshinone isolated from S. miltiorrhiza, which is structurally very similar to cryptotanshinone except C-15 position of dihydrofuran ring does not inhibit angiogenesis induced by bFGF. These results demonstrate that cryptotanshinone is a new anti-angiogenic agent and double bond at C-15 position of the dihydrofuran ring plays a crucial role in the activity. PMID- 15886528 TI - HCV core protein promotes liver fibrogenesis via up-regulation of CTGF with TGF beta1. AB - Liver cirrhosis is one of the major complications of hepatitis C virus (HCV) infection, but the mechanisms underlying HCV-related fibrogenesis are still not clear. Although the roles of HCV core protein remain poorly understood, it is supposed to play an important role in the regulation of cellular growth and hepatocarcinogenesis. The aim of this study was to examine the role of HCV core protein on the hepatic fibrogenesis. We established an in vitro co-culture system with primary hepatic stellate cell (HSC) isolated from rats, and a stable HepG2 HCV core cell line which had been transfected with HCV core gene. The expressions of fibrosis-related molecules transforming growth factor beta1 (TGF-beta1), transforming growth factor beta receptor II (TGFbetaRII), alpha-smooth muscle actin (alpha-SMA) and connective tissue growth factor (CTGF) were analyzed via histological or molecular methods. In addition, the expression levels of matrix metaloprotinase-2 (MMP-2) and collagen type I (Col I) from the co-cultured media were measured by zymogram and ELISA, respectively. The expressions of alpha-SMA, TGF-beta1, Col I, TGFbetaRII and MMP-2 were significantly increased in the co culture of stable HepG2-HCV core with HSC. Moreover, the significant increases of CTGF and TGF-beta1 in the HCV core-expressing cells were observed by either Northern or Western blot analysis. These results suggest that HCV core protein may contribute to the hepatic fibrogenesis via up-regulation of CTGF and TGF beta1. PMID- 15886529 TI - Single-pill therapy in the treatment of concomitant hypertension and dyslipidemia (the amlodipine/atorvastatin gemini study). AB - The Gemini Study was a 14-week, open-label, non-comparative, office-based, multicenter trial to evaluate single-pill therapy in the treatment of concomitant hypertension and dyslipidemia. In addition to recommending lifestyle modifications, eight dosage strengths of amlodipine/atorvastatin single pill (5/10, 5/20, 5/40, 5/80, 10/10, 10/20, 10/40, and 10/80 mg) were electively titrated to improve blood pressure and lipid control. A total of 1220 patients with uncontrolled hypertension at baseline received study medication. At baseline, mean blood pressure was 146.6/87.9 mm Hg and mean low-density lipoprotein cholesterol concentration was 152.9 mg/dL. At study end, 57.7% of patients had achieved both their blood pressure and low-density lipoprotein cholesterol goals (51.9% of patients with uncontrolled low-density lipoprotein cholesterol at baseline). The mean dose of study medication at end point was amlodipine component 7.1 mg and atorvastatin component 26.2 mg. Fifty-eight patients (4.8%) discontinued therapy due to adverse events. Single-pill therapy is effective in reducing both blood pressure and lipid levels and in helping patients achieve goals for both hypertension and dyslipidemia. PMID- 15886530 TI - Quality of life and antihypertensive drug therapy. AB - Quality of life on antihypertensive therapy is an important consideration because clinicians are asked to initiate drug therapy and follow mostly asymptomatic patients for long periods of time on agents that are fairly equivalent in both blood-pressure-lowering capacity and the reduction of adverse clinical events. There is, however, evidence to show that hypertension is not always an asymptomatic condition; therefore, the reduction of blood pressure makes people not previously knowledgeable of their hypertensive state feel better. Labeling a patient hypertensive may have negative quality-of-life consequences. Clinicians need to be well informed regarding side-effect profiles as well as anxiety conditions that may lead to subjective complaints that are blamed on medication. Additionally, medication information given to patients may have an important effect on adverse effect reporting. Specific intolerance profiles to the thiazides, angiotensin-converting enzyme inhibitors, beta blockers, calcium channel blockers, and angiotensin receptor blockers are discussed in this review. Medication compliance requires a multi-tiered strategy. Low-dose thiazide is well tolerated. PMID- 15886531 TI - Stroke in patients with diabetes and hypertension. AB - Stroke is major public health problem leading to increased morbidity and mortality. Modifiable risk factors for stroke include hypertension, diabetes, atrial fibrillation, dyslipidemia, smoking, and alcohol abuse. Among these risk factors, diabetes and hypertension are rapidly growing epidemics leading to a substantial increase in cardiovascular disease and stroke. In this review, the authors discuss the risk factors for stroke with emphasis on the diabetic and hypertensive population, highlighting the interventions that have been shown to decrease stroke risk in this patient population. PMID- 15886532 TI - Roundtable discussion: the role of Beta blockers in the management of hypertension. AB - Following a hypertension symposium in New York City on February 2, 2005, a roundtable was convened to discuss the use of beta blockers in the management of hypertension. Dr. Marvin Moser, Clinical Professor of Medicine at the Yale University School of Medicine, New Haven, CT, moderated the discussion. Participating in this roundtable were Dr. Thomas Pickering, Professor of Medicine at the Columbia University School of Medicine, New York, NY, and Dr. William Cushman, Professor of Preventive Medicine and Medicine at the University of Tennessee College of Medicine, Memphis, TN. PMID- 15886533 TI - Gender and blood pressure. AB - The prevalence, impact, and control of hypertension differ between the sexes in the US population. In addition, pregnancy, oral contraceptive use, and menopausal hormone therapy may influence blood pressure regulation in ways that have therapeutic implications for some women. Whether gender should be a significant consideration in the choice of individual antihypertensive drugs continues to be a topic of intense interest and debate. This brief review will discuss recent findings that bear on considerations of gender in the management of hypertension, particularly among older women. PMID- 15886534 TI - Why don't we pay more attention to the effects of stress in our patients? PMID- 15886537 TI - Assessment of age-related changes in left ventricular structure and function by freehand three-dimensional echocardiography. AB - To determine age-related changes in left ventricular (LV) structure and function, the authors used freehand three-dimensional echocardiography, a previously validated tomographic technique, to measure LV mass, volumes, and derived parameters in 94 sedentary, but ambulatory and clinically healthy, adult male and female volunteers aged 20-94. LV volumes and mass were significantly greater in men than in women (p<0.001) and remained greater after adjustment for body surface area and height (p<0.05). Declines in end-diastolic and stroke volumes with age were reduced or absent after accounting for body size. By multivariate analysis, age accounted for only about 6%-11% of the variance among LV volumes and mass, in comparison with body surface area and gender, which accounted jointly for about 46%-77% of the variance. In conclusion, changes occur in LV structure and function with normal aging, but these changes are relatively minor when body size and gender are taken into account. PMID- 15886538 TI - JNC 7 category and risk of cardiovascular death in men: are there differences by age? AB - The authors evaluated the seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) blood pressure classification scheme on the risk of cardiovascular disease death and whether the association varies by age. In a prospective cohort study, 53,163 apparently healthy participants aged 39-85 years were followed for 5.7 years. The authors calculated relative risks and 95% confidence intervals for cardiovascular disease death, adjusting for major risk factors, then stratified by age (39-49, 50-59, 60-69, and 70-84 years) and performed a test of heterogeneity. Compared with men with normal blood pressure, those with prehypertension had a multivariate-adjusted relative risk of 1.02 (95% confidence interval, 0.73-1.42) and those with hypertension had a relative risk of 1.54 (95% confidence interval, 1.08-2.19). There was no strong age-related effect (P(interaction), 0.22). In this large cohort, the JNC 7 categorization of prehypertension was not associated with a significantly increased risk of cardiovascular disease death, and there was no significant effect modification by age. PMID- 15886539 TI - Heart failure in the oldest patients: the impact of comorbid conditions. AB - Heart failure (HF) is the leading cause of hospitalization in older adults; it is also an important cause of death and chronic disability. HF in the elderly differs in many respects from HF occurring during middle age; in particular, the diagnosis and treatment of HF in the elderly are often complicated by the presence of multiple cardiac and noncardiac comorbid conditions, many of which have important implications for the care of the older HF patient. This article reviews the effects of common noncardiac comorbidities on the management of HF in older adults and discusses the impact of noncardiac comorbid conditions on clinical outcomes in the geriatric HF patient. PMID- 15886540 TI - Endovascular therapies for noncoronary atherosclerosis in the elderly: supra aortic vessels and thoracoabdominal aorta lesions. AB - Due to the aging of the population and the fact that people are living longer, there are increasing numbers of older patients with noncoronary atherosclerosis. Carotid and subclavian arteries are often the first involved vessels in atherosclerosis, and thoracoabdominal aortic involvement is becoming even more frequent. New techniques, such as intra-arterial thrombolysis, protection and thrombo-aspiration devices, and new stent graft designs are becoming available for percutaneous treatment of atherosclerosis in such vessels. In this review, the authors offer geriatric cardiologists an overview and an update of the most recent advances in techniques and results in the field of interventional treatments of atherosclerosis of supra-aortic vessels and thoracoabdominal aorta in the elderly. PMID- 15886541 TI - Ethical issues in the management of geriatric cardiac patients. AB - Ethical Issue: A hospital's Ethics Committee decides to not give analgesics to a terminally ill patient to relieve her pain. PMID- 15886542 TI - Outcomes of regional variation in invasive vs. medical management of patients with acute myocardial infarction. PMID- 15886543 TI - Alcohol--a cardiovascular drug? PMID- 15886544 TI - Saphenous vein graft aneurysm presenting as cardiac mass on echocardiography. PMID- 15886545 TI - Multifocal atrial arrhythmia. PMID- 15886547 TI - What's wrong with me? Women's coronary heart disease diagnostic experiences. AB - Most women are unaware that that they may experience atypical coronary heart disease (CHD) symptoms. Women's atypical presentation often results in women having difficulty being diagnosed with CHD or myocardial infarction. Investigating women's CHD diagnostic experiences may reveal vital areas amenable to intervention. This secondary analysis explored women's CHD diagnostic experiences. Forty women completed in-depth interviews in their homes that were audiotaped and lasted 2-3 hours. Using content analysis and constant comparison, five themes emerged: awareness, seeking treatment, frustration, treatment decisions, and anger. Despite numerous symptoms and visits with clinicians, most women were not diagnosed with CHD before myocardial infarction. During the infarction, women with typical symptoms were easily diagnosed while those with atypical symptoms received a delayed diagnosis. Those who repeatedly sought treatment were angry about not being diagnosed earlier. Further research is needed to promote early symptom recognition, timely diagnosis, and efficacious treatment-keys to improving women's CHD outcomes and to preventing similar negative diagnostic experiences. PMID- 15886548 TI - Gender differences in recovery after coronary artery bypass graft surgery. AB - This study examined the effect of gender on symptom evaluation, symptom response, and physical functioning following coronary artery bypass graft surgery. Symptom evaluation and symptom response were measured at baseline and 2, 4, and 6 weeks and 3 months following surgery, and physical functioning was measured at baseline, 6 weeks, and 3 months. The sample included 46 men and 19 women randomly assigned to the routine care group of a larger study examining symptom management intervention influence on recovery outcomes in elderly coronary artery bypass graft patients. No significant differences were found in symptom evaluation, symptom response, or physical functioning by gender over time. Women reported consistently higher symptom evaluation scores for shortness of breath, fatigue, depression, sleep disturbance, swelling, and anxiety and lower mean physical and vitality subscale scores than men at every time point. Gender-specific symptom management strategies should be developed and tested to address the differences in the reporting of symptoms by men and women. PMID- 15886549 TI - Cardiovascular effects of coffee: is it a risk factor? AB - Intake of coffee, one of the most common beverages worldwide, is often reported as a cardiovascular risk factor; however, definitive data are lacking. Acute intake of coffee or beverages containing caffeine can increase blood pressure, heart minute volumes, and cardiac index, as well as activate the sympathetic nervous system in nonhabitual coffee drinkers. Interestingly, this is not observed in habitual coffee drinkers. Restriction of coffee or caffeinated beverages is no longer indicated in the seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) guidelines for the treatment of hypertension. In fact, no clear association between coffee and the risk of hypertension, myocardial infarction, or other cardiovascular diseases has been demonstrated. In contrast to early studies, recent research indicates that habitual moderate coffee intake does not represent a health hazard and may even be associated with beneficial effects on cardiovascular health. PMID- 15886550 TI - The Mediterranean diet: is it cardioprotective? AB - Coronary heart disease is one of the leading causes of morbidity and mortality in the United States. Dietary interventions are first-line therapy for coronary heart disease prevention and treatment. Increasing scientific evidence suggests that the traditional Mediterranean diet may reduce the risk of cardiovascular disease. The cardiovascular benefits of this whole-diet approach may outweigh those of typically prescribed low-fat diets. The burden of coronary heart disease is enormous, and nutritional approaches that optimize cardiovascular health are essential. Clinical trial evidence supporting the role of the Mediterranean diet in cardiovascular health is presented with an emphasis on the physiological effects of omega-3 fatty acids. Implications for clinical practice and future research are also discussed. PMID- 15886551 TI - Cardiovascular nursing in Spain. PMID- 15886552 TI - Practical challenges in the management of oral anticoagulation. PMID- 15886553 TI - Infection: an Achilles heel of destination left ventricular assist device therapy. PMID- 15886555 TI - Is there an output failure? PMID- 15886556 TI - [What is tropical dermatology? Rare disorders are exceptional and common disorders... are common]. PMID- 15886557 TI - [The prospective multicenter study on standard photopatch tests by the French Society of Photodermatology from 1991-2001]. AB - INTRODUCTION: To develop a standard panel of photopatch tests, the French Society of Photodermatology conducted a prospective study from 1991 to 2001 on the frequency of photoallergens encountered in France and on the relevance of the choice of the various photoallergens. PATIENTS AND METHODS: Thirteen photobiology centers participated in the study from 1991 to 1995, and ten centers from 1995 to 2001. A set of 3 samples of photopatch tests was applied on any patient suspected of photoallergy. On Day 2, two sets were irradiated with ultraviolet A (UVA) and total spectrum (DEM 0.75); with the third set being used as control. Readings were made on D3 and D4. RESULTS: Two thousand sixty-seven patients were tested. Eight hundred fifty-six, i.e., 41% exhibited one or several positive tests. In the majority of cases it was a photoallergy (39.7 to 60% of cases) and eczema (29.5 to 45.6%). Photoaggravation was infrequent (7.9 to 10.3%). Cases of phototoxicity were rare. Sesquiterpenic lactones constantly provoked photoallergy, with 12 cases in 10 years. Although phenothiazines were the most photosensitizing allergens up until 1995, they were then overridden by ketoprofen in 1996 with 107 cases of UVA photosensitive reactions (75 cases) and total spectrum (32 cases). These were followed closely by sun screens, benzophenone (notably Eusolex 4360 with 54 pertinent cases of photoallergy) and dibenzoylmethane (with 31 cases due to Eusolex 8020). UVB filters were all potentially photosensitizing but to a lesser degree from 1 to 5 cases). DISCUSSION: Our results differ from those of Anglo-Saxon teams in the appearance of a new photoallergen, ketoprofen, which provoked numerous photosensitivities in both UVA and UVB. This justifies the systematic addition of this substance in our prospective set. Photoallergy was relatively rare, with around 100 cases reported within 10 years. Total spectrum irradiation of the photopatch tests revealed photoallergies that would not have been found with UVA alone. CONCLUSION: Study of photopatch tests has permitted the uniformization of the methodology in France, an overview of the frequency of the photoallergens tested and the development of a new standard set. PMID- 15886558 TI - [Quality of life and pemphigus]. AB - INTRODUCTION: Pemphigus is a frequent chronic bullous dermatosis in Morocco that predominantly affects young women. It requires serious therapeutic management and often repeated hospitalizations. The purpose of this study was to assess the particular impact of pemphigus on quality of life by comparing the quality of life in patients suffering from pemphigus with that of the general population using a generic instrument of measure of health related quality of life: the SF 36, in association with a questionnaire exploring the impact of the disease on self perception, social relationship and behaviour. PARTICIPANTS AND METHODS: A French version of the SF-36 underwent 2 translations from French to Moroccan dialect, then two retrograde translations. After a pre-test, the questionnaire survey was administered by the same investigator to 30 patients who suffered from pemphigus, followed-up in the Ibn Rochd UHC in Casablanca and to 60 healthy adults. The two samples were paired according to age and sex. RESULTS: In the pemphigus group, there was a significant decrease of mean scores of all the SF-36 dimensions, except for physical pain and alteration in general status of health. The greatest alteration concerned the impact on physical and emotional status (p<0.00001) and the smallest alteration concerned the perception of general health (p=0.02). The factors influencing this impact were: profession, face involvement and extent of lesions. Other results concerning the psychosocial impact of pemphigus are presented. CONCLUSION: Our study confirmed that pemphigus is responsible for great alterations in health related quality of life. Therefore, the management of this disease must take into account its impact on various fields of life of the patients. PMID- 15886559 TI - [Ophthalmologic prevention of chloroquine and hydroxychloroquine induced retinopathy]. AB - INTRODUCTION: Antimalarial drugs induce severe retinal toxicity. The aims of this study were to evaluate the strategy of screening clinical and preclinical intoxication due to antimalarial agents in two centres of reference and to describe the results of ophthalmologic examination. PATIENTS AND METHODS: Patients referred for ophthalmologic evaluation in connection with antimalarial agents therapy in the Centre Hospitalier National d'Ophtalmologie des Quinze Vingts from October 1999 to December 2000 and in the Hopital Lariboisiere from January 1995 to December 1998 were investigated. A retrospective review of results of ophthalmologic examination, electroretinogram, electro-oculogram, colour vision test and central visual field was conducted to assess retinal intoxication. RESULTS: Among 705 patients recruited in the Centre des Quinze Vingts, 10 out of 133 who were never treated had an electrophysiological contra indication to the treatment. Among the 572 other patients, 31 presented with preclinical intoxication (5.4 p. 100) and 8 other patients presented with clinical intoxication. Among 925 patients recruited in the Hopital Lariboisiere, 37 presented with preclinical intoxication (4 p. 100) and four patients presented with clinical intoxication. DISCUSSION: The antimalarial drugs clinical intoxication is rare but nevertheless real. Screening for preclinical intoxication can prevent the evolution toward irreversible retinal intoxication. Diagnosis of preclinical intoxication is established through the confrontation of results of different tests and their evolution. The multifocal electroretinogram remains to be evaluated. CONCLUSION: Ophthalmologic monitoring including funduscopy, should be recommended at least once a year. Visual field seems to become interesting in the screening. Electroretinogram and electro-oculogram remain useful quantitative and obvious tests. A prospective study to assess the optimal way to prevent retinal intoxication is mandatory. PMID- 15886560 TI - [Unusual presentation of infantile myofibromatosis with an ulcered plaque]. AB - INTRODUCTION: Infantile myofibromatosis is a rare fibrovascular-like, isolated or multicentric tumor, occasionally of the bone or an organ and appearing before the age of 2. We report a case of infantile myofibromatosis in a child in an atypical form with a single, ulcerated plaque and having developed after the onset of clusters of papular nodules. OBSERVATION: An infant was seen in consultation because of asymptomatic papules that had developed on the back. The histological examination of a partial biopsy revealed a histiocytofibromatus aspect and led to the diagnosis of clusters of multiple histiocytofibromatous. One year later, the papular nodules had converged, forming a large plaque with ulcerated center. The progressive extension and the absence of healing prompted surgical exeresis and the final diagnosis of myofibromatosis. DISCUSSION: Diagnosis of infantile myofibromatosis is difficult histologically and clinically and relies on a clear anatomoclinical confrontation. The clinical aspects are varied. To our knowledge, myofibromatosis with a single ulcerated plaque has never been reported in the literature before. PMID- 15886561 TI - [Diffuse sebaceous hyperplasia of the face induced by cyclosporine]. AB - INTRODUCTION: Many cutaneous complications have been reported in patients treated with cyclosporine. Alterations of the pilosebaceous follicle are particularly frequent. Hypertrichosis, follicular keratosis, acne and folliculitis are very common. Nevertheless, the occurrence of sebaceous hyperplasia is exceptional. OBSERVATION: A 27 year-old man consulted in February 2003 for a papulous eruption of the face. He was treated by cyclosporine and prednisone since his renal transplantation in 1993. The lesions flowed together on the cheeks, forehead and temples. The histological analysis confirmed the diagnosis of sebaceous hyperplasia. There was a perceptible improvement of the cutaneous state after one month of isotretinoin treatment. DISCUSSION: Sebaceous hyperplasia appears in about 10 p. 100 of patients treated with cyclosporine. This side effect occurs only in men of a mean age of 40 years. An increase in sebaceous gland size is often described, but profuse forms are uncommon. Our case report is exceptional because of the young age of the patient, and the occurrence of diffuse sebaceous hyperplasia that appeared a long time after the introduction of cyclosporine. PMID- 15886562 TI - [Treatment of cutaneous metastases of a squamous cell carcinoma of the leg with topical miltefosine]. AB - INTRODUCTION: Miltefosine (Hexadecylphosphocholine) is a chemotherapy, which when applied locally, has demonstrated efficacy in the treatment of cutaneous metastases of breast cancer. CASE REPORT: A 79 year-old woman developed recurrent in transit cutaneous metastases of a squamous cell carcinoma on her left leg. All lesions completely resolved after topical applications of miltefosine. Tolerance was excellent and the patient remained disease-free 24 months after the end of the treatment. DISCUSSION: Miltefosine, which might prove to be a simple and effective alternative for the usually heavy treatments proposed, warrants further assessment in this context. PMID- 15886563 TI - [Worsening of subacute lupus erythematosus induced by infliximab]. AB - BACKGROUND: Infliximab (Remicade) is an anti-TNF alpha indicated in the treatment of chronic inflammatory rheumatism, notably rheumatoid arthritis. CASE-REPORT: We report the case of a 56 year-old woman who developed severe worsening of an SSA positive subacute lupus erythematosus on initiation of treatment with infliximab for rheumatoid arthritis. DISCUSSION: A review of the literature found 30 cases of drug-induced lupus and listed the autoimmune modifications induced by anti-TNF alpha. This first case of subacute lupus erythematosus, existing before the introduction of treatment and worsening during the latter, emphasizes the risk of developing a severe flare of an autoimmune disease during treatment with anti-TNF alpha. It raises the question of the relative contraindications of anti-TNF alpha in patients with lupus erythematosus. PMID- 15886564 TI - [Chronic meningococcemia]. AB - INTRODUCTION: Chronic meningococcemia is a rare clinical form of invasive Neisseria meningitidis infection. We report 2 cases. OBSERVATIONS: A 39 year-old man and a 42 year-old woman had developed a widespread, fleeting and painful maculopapular cutaneous eruption over the past few weeks, associated with intermittent fever and inflammatory arthralgia. In both cases blood cultures isolated a serogroup B meningococcus that confirmed the diagnosis. Cutaneous histology revealed a non-specific image of leukocytoclastic vasculitis. Treatment with beta lactamin antibiotics was successful after respectively 3 weeks and 12 days. DISCUSSION: This septicemia is characterized by the clinical triad of cutaneous eruption, fever and arthralgia. It must not be mistaken for connectivitis because inappropriate corticosteroid prescription may provoke severe complications. Confirmation of the diagnosis is provided by the blood cultures, which should be repeated. In the case of strong clinical suspicion, the rapid improvement with antibiotics confirms the diagnosis. PMID- 15886565 TI - [Cutaneous CD8+ squamous T-cell bullous lymphoma]. AB - INTRODUCTION: Bullous forms of cutaneous T-cell lymphomas are rare. A new group of cutaneous T-cell lymphomas has recently been identified as a distinct clinicopathological and immunophenotype entity. These cutaneous T-cell lymphomas express a CD8+ phenotype, rarely expressed in other cutaneous T-cell lymphomas. CASE REPORT: We describe a cutaneous CD8+ squamous T-cell lymphoma with polymorphic clinical features, strongly epidermotropic lymphoid infiltrate and spongiosis, classical for this type of lymphoma. DISCUSSION: Bullous lesions in cutaneous T-cell lymphoma should evoke the possibility of a cutaneous CD8+ T-cell lymphoma, once other bullous diseases have been excluded. Spongiosis, rare in other types of T-cell lymphoma, and strongly epidermotropic pleomorphic lymphoid infiltrate are classical histological features. The association of polymorphic lesions, bullas and atypical CD8+ epidermotropic phenotype should evoke this diagnosis even at the early stage. Treatment is difficult and classical chemotherapy often fails. Prognosis is poor with a mean overall survival of 32 months. PMID- 15886566 TI - [Advances in skin aging/photoaging: theoretical and practical implications]. PMID- 15886567 TI - [Lingual tuberculosis associated with disseminated tuberculosis]. PMID- 15886568 TI - [Psoriasis worsened by propafenone]. PMID- 15886569 TI - [Use of non prescription drugs for acute dermatoses. A prospective study of 111 hospitalized patients]. PMID- 15886570 TI - [Peri anal tumor]. PMID- 15886571 TI - [Epidemiology of basal cell carcinomas]. PMID- 15886572 TI - [Keloid and hypertrophic scar]. PMID- 15886573 TI - [To each localization its own technique: tunneling preauricular flap]. PMID- 15886574 TI - [Non ablative laser and pulsed light remodeling for aging skin]. PMID- 15886575 TI - [Impact factor of reviews]. PMID- 15886576 TI - [Verrucous carcinoma]. PMID- 15886577 TI - [Noonan syndrome and Leopard syndrome linked to mutation of the gene PTPN11]. PMID- 15886578 TI - [Trichoblastoma of the scalp]. PMID- 15886579 TI - [Urticaria and angiodema due to itraconazole]. PMID- 15886580 TI - [Fixed drug eruption and carbocysteine]. PMID- 15886581 TI - [Do we have to stop treating acne in summertime?]. PMID- 15886583 TI - Non-invasive ventilation as a weaning tool. AB - Patients with chronic airflow obstruction and difficult weaning from mechanical ventilation are at increased risk for intubation-associated complications and mortality because of prolonged invasive mechanical ventilation. Non-invasive positive-pressure ventilation (NPPV) may revert most of the pathophysiologic mechanisms associated with weaning failure in these patients. Several randomised controlled trials have shown that the use of NPPV in order to advance extubation in difficult to wean patients can result in reduced periods of endotracheal intubation, complication rates and survival. The published data with outcome as a primary variable is nearly exclusively from patients who had pre-existing lung disease. In addition the patients were haemodynamically stable, with a normal level of consciousness, no fever and a preserved cough reflex. It remains to be seen whether NPPV has a role in other patients' groups and situations. The technique is however a usual addition to the therapeutic armamentarium for a group of patients who pose a significant clinical and economic challenge. PMID- 15886584 TI - NIV treatment for acute exacerbation in COPD patients. AB - The authors review the clinical applications of non invasive ventilation (NIV) in patients with chronic obstructive pulmonary disease (COPD), mostly focusing on NIV issues and possible solutions. After briefly reviewing the respiratory mechanics modification during an episode of COPD exacerbation, the authors describe the two crucial technical aspects (choice of the interface and leaks control) that are mostly involved in patient-ventilator interaction. Finally, they briefly review the most important clinical trials on NIV. PMID- 15886585 TI - Ventilator or physician-induced lung injury? AB - One of the most challenging problems in critical care medicine is the acute respiratory distress syndrome (ARDS), the most severe form of acute lung injury (ALI). Evidence from experimental studies suggests that mechanical ventilation can cause or aggravate lung injury. Referred to as ventilator-induced lung injury (VILI), this condition resembles ALI and ARDS, and is difficult to identify in humans because its appearance overlaps the underlying disease, supporting the assumption that mechanical ventilation can extend the severity of pre-existent lung injury. There is increasing laboratory evidence that ventilating ARDS models with relatively low tidal volumes and high levels of positive end-expiratory pressure (PEEP) is clinically beneficial. In 2000, the ARDS Network published reported a reduced mortality (from 40% to 31%) in a mixed population of patients with ALI and ARDS ventilated with half the tidal volume of the control group. However, almost forty years after the first description of ARDS, many investigators and experts in the field still apply essentially the same ventilatory strategy (tidal volume greater than 10 mL/kg body weight and PEEP levels less than 10 cmH2O) as in the original description of ARDS. PMID- 15886586 TI - The Medical Emergency Team: 21st century critical care. AB - In-hospital cardiac arrests, intensive care unit (ICU) admissions and unexpected deaths are commonly preceded by warning signs up to 24 hours prior to the event. As a result, some of these critical events are potentially preventable. Critical care physicians are increasingly familiar with patient care systems; trauma systems have become well established in most health services, and the chain of survival provides a system response to out of hospital cardiac arrests. We now need to build on experience with systems to extend critical care services to all hospital patients at risk, whatever their location and on a continuous basis to prevent these critical events from occurring. In fact, if critical care medicine is to take up the challenge and move forward into the 21st century, we need to engage in a re-orientation from individual to system thinking. We know that the majority of in-hospital cardiac arrests occurring on the general wards represent failures in the system. These events are not the fault of one or two individual practitioners that failed to provide adequate care, but a consequence of organisational factors that result in failures in recognition and response involving more than one department, professional group or area of the hospital. There is also potential to reduce morbidity. Morbidity caused by failure to adequately treat hypoxemia and hypovolemia on the wards, results in preventable cases of renal and respiratory failure, requiring prolonged, uncomfortable and expensive admissions to intensive care, along with the invasive therapy that ICU admission entails. The Medical Emergency Team (MET) system provides a potential solution. PMID- 15886587 TI - The use of positive end-expiratory pressure in the management of the acute respiratory distress syndrome. AB - Clinical and experimental research on the effects of positive end-expiratory pressure (PEEP) has produced a plethora of information during the last two decades. The application of PEEP is expected to increase PaO2; however, it is generally agreed that simply using increased PaO2 as the end point is inappropriate. Four mechanisms have been proposed to explain the improved pulmonary function and gas exchange with PEEP: 1) increased functional residual capacity; 2) alveolar recruitment; 3) redistribution of extravascular lung water; and 4) improved ventilation-perfusion matching. The optimal method of applying PEEP is still controversial. The main effect of augmenting PEEP is maintain recruitment of alveolar units that were previously collapsed. Thus, since tidal volume is distributed to more alveoli, peak airway pressure is reduced and compliance is increased. During acute lung injury, and depending on the severity of lung disease, PEEP can markedly alter the compliance of the lung by alveolar recruitment. The greater the alveolar collapse and pulmonary edema, the more the compliance curve of the respiratory system shifts downward and to the right. As PEEP is applied and alveoli recruited, the pressure-volume curve shifts upward and to the left. Despite its intuitive benefit, there were very few controlled studies of the effects of PEEP on ARDS outcome and no prospective randomised controlled trial of PEEP has been ever carried out in patients with acute lung injury and/ or ARDS to evaluate its efficacy until recently. PMID- 15886588 TI - Performance determinants and flexible ICU organisation. AB - We faced some of the most important aspects of the problem of the appropriateness of ICU resources use, that are the relationship between volume of activity and mortality, the analysis of cost-effectiveness in intensive care medicine, and the monitoring of the human resource use in ICU. For this aim three different surveys were utilized: one at European level, the second at country level and, third, a regional survey. After developing a new measure of volume called ''high-risk volume'', we explored the relationship between outcome and volume, founding that such association was very strong (from 3 to 1719% decrease in ICU/hospital mortality every five extra high-risk patients treated per bed per year), and that an occupancy rate larger than 80% was associated with higher mortality. Therefore, patients in all levels of risk are better treated in high-risk volume ICUs with a reasonable occupancy rate. Analysing cost-effectiveness in intensive care medicine using a national case-mix categorized in different diagnostic groups, we identified brain haemorrhage, ALI/ARDS and surgical unscheduled patients as users a high volume of monetary resources less efficiently, while the scheduled abdominal surgery patients admitted to receive intensive care and patients on the ICU for minor organ support made the best use of the fewer resources spent. Finally, we designed a new approach to measure the rate and appropriateness of nursing resource use in ICU on a daily basis. Testing this approach on a group of general non-specialist ICUs, we found that the method was powerful enough to adequately distinguish between ''over'' and ''under utilization'' and to identify all the theoretical scenarios of nurse/resource utilization. PMID- 15886589 TI - Therapeutic approaches in trauma-induced coagulopathy. AB - Coagulopathy is a phenomenon which is a life threatening complication in the trauma patient who has sustained significant injuries and blood loss. With our increasing understanding of the mechanisms which drive the coagulopathy and the availability of new treatment options, most notably recombinant factor VIIa (rFVIIa), we are now able to treat those patients who have had a massive traumatic haemorrhage with greater efficacy. This paper reviews the current considerations in dealing with patients with trauma-induced coagulopathy and offers a strategy for their management. PMID- 15886590 TI - Thrombolytics in CPR. Current advantages in cardiopulmonary resuscitation. AB - Cardiac arrest carries a very poor prognosis. More than 70% of cardiac arrests are caused by acute myocardial infarction (AMI) or massive pulmonary embolism (PE). Thrombolysis during CPR has two major effects: first, it causally treats the condition that caused cardiac arrest and second, it has been shown to have beneficial effects on the microcirculatory cerebral reperfusion after cardiac arrest. However, this treatment has been widely withheld mainly because of the fear of severe bleeding complications. We reviewed the currently available in- and out-of-hospital studies on thrombolysis during CPR. Most studies found that thrombolytic therapy during CPR improves the chance for a restoration of spontaneous circulation in patients suffering from cardiac arrest and may even result in a better outcome. In addition, the neurological condition of surviving patients may be markedly improved by thrombolysis. Although thrombolytic therapy is associated with a risk of bleeding complications, currently available data do not suggest an increase of bleeding complications if thrombolysis is administered during CPR. Recently, a large randomized multicentre study has started to assess the efficacy and safety of thrombolysis during prehospital CPR. PMID- 15886591 TI - Management of prehospital thrombolytic therapy in ST-segment elevation acute coronary syndrome (<12 hours). AB - Acute myocardial infarction (AMI) is the prototype of a real emergency, and both efficacy and speed are necessary for effective management. The advent of thrombolysis therapy has transformed the care of these patients. In fact, the most frequent complication of AMI is sudden death which still occurs within the first hour after symptom onset. Thrombolytic therapy has been shown to reduce early and long term mortality about 20%. The mortality gain is dependent on the delay time of early reperfusion. A large number of studies have shown that this relationship is best described as exponential: in the first 1 to 2 hours after the onset of chest pain, the benefit of thrombolysis is greater. Reducing the time to thrombolysis must therefore be the main objective of prehospital treatment of AMI. In the last 10 years, a large number of strategies to reduce the time to reperfusion have been evaluated, including initiation of thrombolytic therapy prior to arrival to hospital. In France, prehospital emergency medicine is an integral part of the medical care system. The SAMU is a hospital department whose function is to centralize emergency medical calls and organise an appropriate response with the intention of ensuring the shortest delay between the initial call and the appropriate treatment. In the event of an emergency medical call concerning chest pain, the medical dispatcher of SAMU may decide to send a MICU (mobile intensive care unit). If a diagnosis of AMI is confirmed, clinical ECG criteria, prehospital thrombolysis is currently seen as the best treatment strategy. The SAMU experience has proven that prehospital thrombolysis is both safe and effective. During the last ten years to fifteen years the field of reperfusion during acute myocardial infarction was a real battlefield between the proponents of thrombolysis and those of primary percutaneous interventions. Nowadays there is a growing number of physicians who will consider that the best way forward is not to oppose these two effective methods but to find the most appropriate niche for each or even better to combine them to achieve reperfusion. In this respect, the concept of facilitated percutaneous intervention is a very attractive one which shows promising results. A large number of studies are now ongoing to demonstrate its efficacy and to help us to choosing the ideal combination of anti-thrombotic agents to be used. That is one of the main interests of the CAPTIM study. French trial comparing prehospital thrombolysis to primary angioplasty. There is no difference between the two strategies in term of primary end points. That could be the real life for acute myocardial infarction. We have to consider in this study the fact than 33% of the patients had a pre hospital thrombolysis followed by a fast angioplasty. The results are impressing: the 30 day mortality in the pre hospital thrombolysis arm is only 3.8%. But if the delay between pain to pre hospital thrombolysis is under 2 hours this 30 day mortality fall down to 2.2%. This is better RESULTS: Than il all the recent trials published comparing on site thrombolysis to primary angioplasty (DANAM II, C Port, PRAGUE II). These good results in the CAPTIM study when the delay pain to treatment is less than 2 hours include also the occurrence of cardiogenic shock in favour of pre hospital thrombolysis (1.3%). The good strategy in a next future could be the association of pre hospital thrombolysis and angioplasty. In a recent French register (USIC 2000) including all the patients arriving in CICU during a month and regarding the one month mortality this strategy seems to be the best (3.6%). The arrival of TNK-tPA is now changing the general management of prehospital AMI by reducing the time to treatment. This is clearly now the new standard of prehospital treatment. The reduction of UHF dose is recommended and the LWMH is considered as the next step as recently demonstrated in the ASSENT 3 and 3+ trials. Several recent registries have shown than we offer reperfusion to only half of the patients and even more important, when we do not offer it, this is unjustified in nearly half of the cases and these patients , forgotten for reperfusion have all a very poor prognosis. The other major problem is that patients are treated too late mainly because the call the emergency system too late. The are several ways to improve the time to treatment : information of the patients , shortening of the intra-hospital delays by better organisation and finally and perhaps more importantly , pre hospital triage and treatment. The efficacy and safety of the pre hospital strategy is now recognised worldwide. The best strategy for acute myocardial infarction should involve emergency physicians and cardiologist in a real local task-force to join and coordinate their efforts. That is the way to open more arteries earlier, that is to say save myocardium and more lives. PMID- 15886592 TI - Volumetric monitoring: principles of application. AB - The transpulmonary thermodilution indicator (TPID) technique has been recently introduced and diffuse in clinical practice. This "less-invasive" device measures intermittent cardiac output and, based on pulse contour method, continuous cardiac output, that agree with cardiac output obtained with pulmonary artery catheter in different clinical setting. Moreover it allowed stroke volume variation and pulse pressure variation experimental and clinically validate fluid responsiveness index in controlled mechanically ventilated patients. The TPID technique allowed an estimations of preload index such as intrathoracic blood volume and "lung edema" index as extra vascular lung water. We reviewed the principle of clinical application based on the current literature now available from this device. Cardiac output monitoring based on TPID technique is safe and accurate, as well as fluid responsiveness indicator (SVV and PPV). Intrathoracic blood volume seems to be a good preload index but the results reported in literature are not homogeneous in all its applications. Extra vascular lung water index is a very interesting parameter particularly in critically ill setting but its clinical application is not yet widely documented. PMID- 15886593 TI - Right ventricular failure: physiology and assessment. AB - Right ventricular function can be altered in several disease states involving lungs and heart. Severe right ventricular dysfunction is a major determinant of outcome in such situations, and may strongly influence clinical management. The complex geometry of the right ventricle and the different physiology with respect to the left ventricle make the right ventricular failure difficult to define and assess. The response to increased afterload is the main determinant of right ventricle physiology in pathologic conditions. This consists of right ventricular hypertrophy and enlargement, with reduced coronary blood flow to the right ventricular wall, dilation of tricuspid annulus and displacement of interventricular septum. This latter change involves the left ventricular diastolic function, which is reduced by leftward septal shifting. In right ventricle myocardial ischemia and infarction the primum movens of altered right ventricular function is not an increase in afterload, but the ischemic involvement of the right ventricle, more often in the setting of an inferior acute myocardial infarction. The assessment of right ventricular failure is based on thermodilution by pulmonary artery catheter, contrast and radionuclide ventriculography, echocardiography, and magnetic resonance. Among these techniques, thermodilution and echocardiography play a relevant role in clinical scenarios, being readily available and feasible bedside. PMID- 15886594 TI - Nitric oxide in thoracic surgery. AB - Inhalation of nitric oxide has been reported to alter pulmonary blood flow in animal and human studies. This effect is related to the relaxant action of nitric oxide on arterial vascular smooth muscle cells. When nitric oxide is administered by inhalation, this effect is limited to the pulmonary vasculature as it is rapidly inactivated by hemoglobin as soon as it enters the blood stream. The effect of inhaled nitric oxide is more pronounced in well ventilated areas of the lung, where it promotes redistribution of pulmonary blood flow to regions with high ventilation-perfusion ratio decreasing pulmonary hypertension and improving oxygenation. Nitric oxide has been used to treat pulmonary hypertension and hypoxemia that occurred in thoracic surgery during one lung ventilation, postpneumonectomy pulmonary edema and lung transplantation. Inhaled nitric oxide may be a useful tool in patients with a low PaO2/FiO2 ratio during one lung ventilation. Further powered studies are still required to define the dose and timing of inhaled nitric oxide in patients who do have ischemia-reperfusion injury after lung transplantation. PMID- 15886595 TI - Pulmonary perfusion during anesthesia and mechanical ventilation. AB - Cardiac output and the pulmonary perfusion can be affected by anesthesia and by mechanical ventilation. The changes contribute to impeded oxygenation of the blood. The major determinant of perfusion distribution in the lung is the relation between alveolar and pulmonary capillary pressures. Perfusion increases down the lung, due to hydrostatic forces. Since atelectasis is located in dependent lung regions, perfusion of non-ventilated lung parenchyma is common, producing shunt of around 8-10% of cardiac output. In addition, non-gravitational inhomogeneity of perfusion, that can be greater than the gravitational inhomogeneity, adds to impeded oxygenation of blood. Essentially all anaesthetics exert some, although mild, cardiodepressant action with one exception, ketamine. Ketamine may also increase pulmonary artery pressure, whereas other agents have little effect on pulmonary vascular tone. Mechanical ventilation impedes venous return and pushes blood flow downwards to dependent lung regions, and the effect may be striking with higher levels of PEEP. During one-lung anesthesia, there is shunt blood flow both in the non-ventilated and the ventilated lung, and shunt can be much larger in the ventilated lung than thought of. Recruitment manoeuvres shall be directed to the ventilated lung and other physical and pharmacological measures can be taken to manipulate blood flow in one lung anesthesia. PMID- 15886596 TI - Morbidity and mortality related to anesthesia outside the operating room. AB - Morbidity and mortality related to sedation or anesthesia outside the operating room has not been investigated so far, but it is assumed to be a relevant problem because the increasing needs for sedation/analgesia in remote locations for a wide range of diagnostic and operative procedures (endoscopy, radiology, magnetic resonance...) and the lack of monitoring, inadequate training of personnel,insufficient staffing. Many complications could occur to patients, like anaphylactic shock,accidental hypothermia,difficult airway maintenance, aspiration,nausea and vomiting, and anesthesiologists, like exposure to pollution, radiation, electromagnetic fields, falls and trauma. Recent guidelines and personal experience are presented and discussed. PMID- 15886597 TI - Target controlled infusion: TCI. AB - Progress in computing technology has allowed the development of target controlled infusion devices, with drugs delivered to achieve specific predicted target blood drug concentrations. Target controlled infusion (TCI) system has been developed as a standardised infusion system for the administration of opioids, propofol and other anaesthetics by target controlled infusion. A set of pharmacokinetic parameters has been selected using computer simulation of a known infusion scheme. The selected model is incorporated into a computer-compatible infusion pump. Clinical trials with such systems have provided appropriate target concentrations for the administration of target controlled infusion of anaesthetic drugs. The technique of TCI strongly influences the development of intravenous anaesthesia and opens a scenario of new and exciting applications in peri-operative anaesthetic management. The launch of ''Diprifusor'' as the first commercially available TCI system for propofol was the cornerstone of a successful research period within the last decade, which evaluated the pharmacokinetic foundations of computer assisted intravenous drug delivery. Nowadays TCI technology is becoming a part of routine anaesthesia technique for the practitioner rather than a research tool for specialists and those who are enthusiasts of intravenous anaesthesia. Besides clinical application in anaesthesia, target controlled systems will play a significant role as research tools in the evaluation of drug interactions in anaesthesia and in the development of new control techniques for the administration of sedative and analgesic drugs in the peri-operative period. PMID- 15886598 TI - Central blocks with levobupivacaina in children. AB - Regional anesthesia has become a routine practice in paediatric anesthesia and local anaesthetics are now widely used in infants and children. Although local anaesthetics are generally quite safe and effective, they may produce systemic toxic reactions affecting the heart and brain. Because postoperative analgesia is often the primary justification for regional anesthesia in infants and children, bupivacaine, a long-acting local anaesthetic, is the most commonly used local anaesthetic for paediatric regional anesthesia. Levobupiva-caine has been used in children by caudal injection, by lumbar epidural route for anesthesia during operation, by continuous epidural infusion for pain control after operation and for spinal anesthesia. Levobupivacaine had shown comparable clinical profiles to that of bupivacaine but produced lower incidence of residual motor blockade. Efforts to minimize the risk of complications during caudal anesthesia must be directed towards measures that reduce accidental intravenous and intraosseous injections, reduce the total amount of local anaesthetic used and use drugs with lower toxic potential. In patients under general anesthesia, when using a large amount of local anaesthetic, in case of accidental intravenous infusion, patients receiving levobupivacaine may tolerate larger doses before manifestation of toxicity compared with those receiving bupivacaine. There are clinical situations including prolonged local anaesthetic infusions, use in neonates or small babies, and caudal block, where replacement of bupivacaine with levobupivacaine appears to be safer. PMID- 15886599 TI - Use of levobupivacaine for the treatment of postoperative pain after thoracotomies. AB - Continuous thoracic epidural analgesia with an opiod-local anaesthetic mixture is the most appropriate strategy to control postoperative pain in thoracic surgery. Levobupivacaine, the pure S(-) enantiomer of racemic bupivacaine, has less cardiotoxic and neurotoxic potential but similar anaesthetic properties of its native agent. There are no studies in thoracic surgery that had established the minimal efficient concentration of this anaesthetic when used with an epidural opioid. The advantages of administering opioids in addition to local anaesthetics in the epidural space are the possibility to decrease dose and consequently side effects of each drug and to exploit the documented synergy between these different categories of drugs in producing segmental epidural analgesia. In our departmental study (unpublished data), 2 different concentration of levobupivacaine (Group A: 0.125% and Group B: 0.0625%) combined with sufentanil (1 mg/mL) were administered in continuous epidural post-thoracotomy infusion to investigate quality of analgesia, motor block and side-effects. An intravenous PCA system has been used in the postoperative period to evaluate rescue morphine consumption. Preliminary results showed that patients of each group reported similar VAS at rest although a better pain control during cough resulted in group A. Patients receiving levobupivacaine at 0.125% presented low incidence of nausea, vomiting and pruritus probably because of the smaller amount of rescue morphine administered. At the concentration of 0.125% epidural levobupivacaine in combination with sufentanil allowed to obtain a good pain control with no adverse effects and motor block at all. PMID- 15886600 TI - Neurologic deficit after peripheral nerve block: what to do? AB - Regional nerve injury is a recognized complication of plexus anesthesia. Nerve damage after regional anesthesia is the most frightening complication for the patient, the anesthetist and the surgeon too. In order to limit possible medico legal issues, the anesthetist has to follow an algorithm. The indication and information obtained from the ultrasonography, the sudomotor test, the electromyogram, the elctroneuromyogram and the evoked potentials are discussed. The principles to avoid anesthetic-related nerve damage during regional anesthesia are given. Finally treatment and surveillance, after nerve damage has occurred, are explained. PMID- 15886601 TI - Local anesthetics in pediatric anesthesia: an update. AB - For several decades, bupivacaine has enjoyed great clinical popularity as the first choice local anesthetic in pediatrics. However, introduction of ropivacaine and levobupivacaine, the two newer and presumably safer (thought to be less cardio- and neurotoxic than bupivacaine) local anesthetics, has challenged the position of bupivacaine as a gold standard for prolonged duration of analgesia. This review article summarized the metabolism, the criteria of choice and the safety use of new agents. PMID- 15886602 TI - Anesthesia for MRI in the paediatric patient. AB - The magnetic resonance imaging suite is a challenging environment for the anaesthesiologists, and carries inherent risks. Several factors account for this, including the remote location, the unique features of the magnetic resonance imaging scanner and patient-related factors. A systematic approach, similar to that of anesthesia provided in the operating room (i.e. proper fasting, informed consent, focused airway examination, medical and surgical history, family history, previous sedation experiences) is mandatory. Understanding the implications of the magnetic resonance imaging environment will facilitate ensuring the safety of the patient. A well-equipped anesthesia machine, standard monitoring (electrocardiogram, oxygen saturation and non-invasive blood pressure), trained personnel and adequate planning should be standard for all out of the operating room procedures. Finally, rigorous discharge criteria are recommended to detect residual sedation. PMID- 15886603 TI - Anesthesiologic preoperative evaluation of drug addicted patient. AB - Even if there are few scientific data about preoperative management of drug addict patients, a careful anesthesiology evaluation facilitates the prevention of interactions between drugs and anaesthetics and avoids intraoperative and postoperative complications. Moreover it is important to earn the confidence of patient in order to know the abuse substances consumed and the frequency of consumption. This knowledge is necessary to the anaesthetist in order to manage possible withdrawal syndromes or overdose, which are the two greatest dangers for a drug addict patient during the hospital stay. PMID- 15886604 TI - Use of protein C concentrate in pediatric patients with sepsis. AB - AIM: Protein C (PC) is a plasma glycoprotein implicated in modulating coagulation and inflammation. Its levels decrease in sepsis and related diseases, where it has also proved to be a prognostic indicator of outcome. Infusion of exogenous PC, although not able to decrease mortality in severe sepsis and septic shock, can safely resolve the coagulation imbalances related to these pathological states. METHODS: A retrospective study was performed about utilisation of PC in severe sepsis and septic shock patients in three italian PICUs during a one-year period. Data from 29 patients were analysed. Age, PIM 2, mortality and length of stay were compared between treated and non treated patients. Treated patients were also analysed for PC dosage received, length of treatment, and modification of hemocoagulation parameters, before PC infusion and every 24 hours. RESULTS: In treated patients, the activity of PC, PT and PTT activity and fibrinogen improved significantly from basal to day 5 (p<0.05). Diminution of d-dimer was not quite significant (p=0.0514). Rise in platelets count and antithrombin III activity was not significant. No adverse reactions related to Protein C concentrate were observed. No difference in mortality was observed between the two groups. CONCLUSIONS: Although PC is included in guidelines for management of severe sepsis and septic shock, only 38%, of observed patients received PC treatment. Even in the treated group, patients received a lower dosage of PC, and for a shorter period, than recommended. In accordance to previous studies, we did not observe differences in mortality between treated and untreated patients. Our results showed a significant increase in plasma PC activity, following infusion of PC concentrate. This increase in PC appeared sufficient to restore some, but not all, of the abnormalities in the coagulation system. A large randomized, phase 3, placebo-controlled trial in children with severe sepsis and septic shock is advisable to establish effective role of therapy with PC in reducing mortality of these patients. PMID- 15886605 TI - Thermal regulation and intraoperative hypothermia. AB - Thermal disturbances are very common perioperatively, especially if paediatric patients are involved. This article consider some aspects of hypothermia during and after surgery under anaesthesia in paediatric patients and how to prevent such disturbances. PMID- 15886606 TI - Infectious diseases of childhood and their anesthetic implications. AB - In western world, infectious diseases in childhood have dramatically decreased in recent years. The first reason is related to the better socio-economic conditions but the highly efficiency of immunizations programs cannot be forgotten. Nevertheless children can be still exposed to infections, as vaccines are not able to completely protect all treated patients. Anesthesiologists should be aware of the basic mechanism of immunization as it is well known that anesthesia and surgery themselves reduce human immune response. In fact, nitrous oxide depresses bone marrow function, while halothane, nitrous oxide again and isoflurane reduce neutrophil biocidal activity. On the contrary, all the anesthesia techniques (peripheral anesthesia) which inhibit stress responses have beneficial effect on the immune system function. Not urgent procedures requiring anesthesia should be deferred for three weeks after vaccination when all the related symptoms will be over. Also important is the knowledge of the incubation period that is the time from contact with a person affected by the infectious illness until the onset of the typical disease. During these periods elective anesthesia must not be performed. On the other hand, if anesthesia is delivered, the patient's ability to react can be compromised and it is possible to have florid disease exacerbation including related complications. PMID- 15886612 TI - Three-dimensional echo: transition from theory to real-time, a technology now ready for prime time. PMID- 15886613 TI - Endoscopic ultrasound for thoracic malignancy: a review. AB - Accurate cancer staging is critical in providing the most appropriate therapy for patients with lung cancer. The decision to attempt a curative surgery or avoid an unnecessary surgery is dependent on accurate staging. In the case of non-small cell lung cancer (NSCLC), the most important parameters for optimal treatment and prognosis are the presence of cancer spread to the lymph nodes within the mediastinum and to distant organs. Endoscopic ultrasound (EUS) has become an important tool for the assessment of mediastinal lymph nodes and in some cases, distant organ metastases, because of its minimally invasive access to these sites through a transesophageal, transgastric, or transduodenal approach. The capability of performing fine needle aspiration (FNA) has greatly improved the accuracy and acceptability of EUS for lung cancer staging. This review will outline the basic principals of EUS-guided lung cancer staging and EUS-FNA techniques and outline the indications and contraindications to EUS staging of thoracic malignancy. PMID- 15886614 TI - Intermediate cystic renal masses: radiology-pathology correlation. PMID- 15886615 TI - Consortium of otolaryngology-head and neck surgery journals to collaborate in maintenance of high ethical standards. PMID- 15886616 TI - Seldinger-assisted videotelescopic intubation (SAVI): a common sense approach to the difficult pediatric airway. AB - OBJECTIVES: To describe the Seldinger-assisted videotelescopic intubation (SAVI) technique, a complementary method for aiding in difficult pediatric intubations that uses common equipment available to the practicing otolaryngologist. STUDY DESIGN: Technique description. METHODS: Detailed description of technique for use of a pediatric laryngoscope with video-assisted endotracheal tube (ETT) covered rigid tracheoscopy controlled intubation in difficult pediatric airways. RESULTS: In our practice, SAVI is vital in establishing a secure airway in the difficult to-intubate child. After insertion of a laryngoscope, an appropriate sized endotracheal tube is delivered through the glottis under direct video visualization from a rigid telescope using a variation of the well-established Seldinger technique. The telescope serves as the stable ETT stylet that also provides panoramic visualization. The ETT slides over the telescope to provide a secure airway directly visible to all in the operating room. Benefits of the SAVI technique to previously described video-assisted intubations with flexible or specially designed endoscope devices include decreased cost, employment of previously existing endoscopy skills, the benefit of rigid delivery of the endotracheal tube as well as innate versatility to a multitude of clinical situations. CONCLUSIONS: The SAVI technique offers an additional practical clinical solution to the difficult pediatric airway. Although ultimately establishing the airway depends on the skills of the operator, the SAVI technique has saved multiple lives by using common equipment through a common-sense approach. EBM RATING: D. PMID- 15886617 TI - Outcome of adenotonsillectomy for obstructive sleep apnea in children under 3 years. AB - OBJECTIVE: To study the outcome of adenotonsillectomy for obstructive sleep apnea (OSA) in children less than 3 years of age. DESIGN AND SETTING: Prospective study at the University of New Mexico Children's Hospital. Children with OSA underwent pre- and postoperative full-night polysomnography (PSG). Scores were compared using a paired t test. A P -value <0.05 was considered significant. RESULTS: The study population included 20 children. Fifteen (75 %) were male. The mean age was 2.2 years (range, 1.1 to 3.0). Sixteen (80%) children had medical comorbidities. Over 25% of children had postoperative complications including laryngospasm and marked desaturations. The mean preoperative respiratory distress index (RDI) was 34.1 and the mean postoperative RDI was 12.2 ( P < 0.0001). After surgery, 7 (35%) children had an RDI < 5. Thirteen (65%) had a postoperative RDI > or = 5 indicating persistent OSA. CONCLUSION AND SIGNIFICANCE: Children under 3 years show significant improvement in RDI after adenotonsillectomy for OSA, but they may develop complications after surgery. Postoperative PSG is recommended for children under 3 years of age to monitor the severity of persistent OSA. EBM RATING: B-2. PMID- 15886618 TI - Long-term histological outcomes of injected autologous fat into human vocal folds after secondary laryngectomy. AB - OBJECTIVE: To histologically evaluate the long-term outcomes of autologous fat grafts after injection laryngoplasty in the human larynx. STUDY DESIGN AND SETTING: We injected liposuctioned fat for vocal fold augmentation in patients with vocal fold paralysis. We suctioned autologous fat from the low abdomen with an 18-G disposable needle and a 20-mL disposable syringe under negative pressure. This is different from the conventional liposuction technique and avoids the use of special equipment. In this article, we report the histological evaluation of 2 patients (patient 1: 12 months, patient 2: 41 months) who required total laryngectomy after autologous fat injection into the vocal folds. RESULTS: Histological examination revealed normal-appearing viable adipocytes with minimal inflammatory response in both patients. CONCLUSIONS: Our liposuctioned autologous fat injection histologically offered long-term improvement in patients with impaired glottal closure from vocal fold paralysis. PMID- 15886619 TI - Adult acute epiglottitis: experiences in a Taiwanese setting. AB - OBJECTIVE: Acute epiglottitis is a potentially disastrous disease seen occasionally in otolaryngology emergency service. This study reports our experiences in managing adult patients with acute epiglottitis in a Taiwanese setting. STUDY DESIGN: Admission medical records from 46 adult patients with acute epiglottitis over a period of 8 years in a tertiary referral otolaryngology service were retrospectively reviewed. RESULTS: No mortality was reported; 4 (8.7%) patients were supported by artificial airway. A total of 16 (34.8%) patients had comorbid conditions; hypertension and diabetes mellitus were the most common systemic diseases. Patients who needed artificial airway support tended to have more systemic comorbidities ( P = 0.001) and a higher level of hyperglycemia ( P = 0.004) than those who did not need airway support. The incidences of drooling, stridor/dyspnea, and muffled voice were significantly higher in the airway-supported group ( P < 0.05). Stridor/dyspnea is a reliable clinical predictor of airway compromise (odds ratio 2.94; P = 0.0277). CONCLUSION: Dyspnea is a warning sign of impending airway collapse during an episode of acute epiglottitis. Adult patients with more systemic comorbidities might bear higher risk of airway compromise; the role of diabetes mellitus can't be overlooked. PMID- 15886620 TI - Antigen challenge induces a supraglottic but not a subglottic edema in the rat larynx. AB - OBJECTIVE: To examine the reaction of mast cells in the larynx of sensitized rats after allergen challenge and determine whether a type I hypersensitivity reaction can elicit a supraglottic and subglottic edema. STUDY DESIGN: Rats were sensitized with ovalbumin (OA) and challenged 3 weeks later by intravenous injection of either OA or NaCl and perfused with glutaraldehyde. The larynges were removed and processed for light and electron microscopy. Sections from the epiglottis and subglottic region were examined. RESULTS: In challenged animals, the connective tissue mast cells (CTMC) in the epiglottic submucosa, but not the mucosal mast cells (MMC) in epithelium, were significantly reduced compared with the case of controls, and the connective tissue was edematous. In the subglottic region, the number of both MMC and CTMC was reduced (albeit not significantly), and there was no edema in the lamina propria. CONCLUSIONS: Allergen challenge induces a marked edema in the epiglottis but not in the subglottic area. SIGNIFICANCE: The results indicate that the supraglottic and subglottic regions react differently to allergen challenge. PMID- 15886621 TI - Intrinsic laryngeal muscle reinnervation with nerve-muscle pedicle. AB - OBJECTIVE: To test the application of the nerve-muscle pedicle (NMP) technique for selective reinnervation of previously denervated posterior cricoarytenoid (PCA) muscle. METHODS: The left recurrent laryngeal nerve (RLN) was severed in 5 mongrel dogs, and an ansa cervicalis-sternohyoid muscle pedicle was sutured to the left PCA muscle. Three dogs underwent a sham operation. Videolaryngoscopy was performed, and electromyographic data were collected after 1 year on average. Finally, histologic analysis of the NMP was performed. RESULTS: The video records showed the return of mobility of the PCA muscle reinnervated by the NMP. EMG data as to show evoked polyphasic potentials showed also evidence of reinnervation of the PCA muscle. With immunohistochemical reaction (antineurofilament antibody+biotin) we could show neurofilaments and motor endplates in both sides in all 5 animals. CONCLUSIONS: The NMP technique could eliminate the need for arytenoidectomy and laterofixation in patients with unilateral or bilateral vocal fold paralysis. The quality of life and voice may be improved. PMID- 15886622 TI - Inpatient management of epistaxis: outcomes and cost. AB - OBJECTIVE: Evaluate treatments for epistaxis. STUDY DESIGN AND SETTING: Retrospective review of Nationwide Inpatient Sample (1998-2000). RESULTS: A total of 9778 admissions with admitting diagnosis "epistaxis" were identified. Among admissions involving 1 treatment, 454 (9.6%) received arterial ligation, 94 (2.0%) embolization, and 4188 (88.4%) nasal packing. There were no differences in length of stay, transfusions, complications, or deaths between groups (all P > 0.05). Mean total hospital charges were USD 6,282 for the packing group, USD 12,805 for the ligation group, and USD 17,517 for the embolization group; differences between ligation and packing groups, and embolization and packing groups demonstrated significance ( P < 0.05). CONCLUSIONS: Nasal packing is used commonly for epistaxis that requires inpatient management. Although embolization and arterial ligation are associated with higher hospital charges, complications, transfusion rates, and lengths of stay are similar. Further studies are needed to quantify other outcome measures, such as recurrence rates and patient quality of life. SIGNIFICANCE: Nasal packing is associated with lower hospital charges and similar complication rates as arterial ligation or embolization. PMID- 15886623 TI - Nitric oxide level in the nasal and sinus mucosa after exposure to electromagnetic field. AB - OBJECTIVE: The purpose of this study was to examine the changes in nitric oxide (NO) level in the nasal and paranasal sinus mucosa after exposure radiofrequency electromagnetic fields (EMF). STUDY DESIGN AND SETTING: Thirty male Sprague Dawley rats were randomly grouped as follows: EMF group (group I; n, 10), EMF group in which melatonin received (group II; n, 10) and the control (sham operated) group (group III; n, 10). Groups I and II were exposed to a 900 MHz. Oral melatonin was given in group II. Control rats (group III) were also placed in the tube as the exposure groups, but without exposure to EMF. At the end of 2 weeks, the rats were sacrificed, and the nasal and paranasal sinus mucosa dissected. NO was measured in nasal and paranasal mucosa. RESULTS: The nasal and paranasal sinus mucosa NO levels of group I were significantly higher than those of the control group (group III) ( P < 0.05). However, there was no statistically significant difference between group II and the control group (group III) regarding NO output ( P > 0.05). CONCLUSION: Exposure to EMF released by mobile phones (900 MHz) increase NO levels in the sinus and nasal mucosa. SIGNIFICANCE: Increased NO levels may act as a defense mechanism and presumably related to tissue damage. In addition, melatonin may have beneficial effect to prevent these changes in the mucosa. PMID- 15886624 TI - Oropharyngoesophageal scintigraphy in the diagnostic algorithm of laryngopharyngeal reflux disease: a useful exam? AB - OBJECTIVE: To evaluate the role of oropharyngoesophageal scintigraphy in the diagnostic approach to patients with laryngopharyngeal reflux (LPR). STUDY DESIGN: Forty-one patients with chronic laryngopharyngeal symptoms and a control group of 15 healthy volunteers were examined. All subjects underwent standard oropharyngoesophageal scintigraphy. The following parameters were considered: activity-time curves, presence of double peaks and reduced slope in oropharyngeal phase, presence of accelerated or slowed esophagogastric transit, persistence of radioactive material on the pharyngeal or esophageal mucosa, and presence of gastroesophageal reflux (GER) under exercise. RESULTS: There were objective signs of laryngeal reflux in 80.5% of the patients. In 82.9%, scintigraphy documented several associated morphofunctional pathologic patterns: positive GER test (61.0%), presence of double peaks (36.6%), indirect signs of pharyngoesophageal inflammation (31.7%), hypotonic lower esophageal sphincter (17.1%), and slowed esophageal clearance (9.8%). CONCLUSIONS: Oropharyngoesophageal scintigraphy may be used as preliminary examination in outpatients with signs and symptoms of LPR. Cases of classic gastroesophageal reflux disease could be separated from those of LPR disease and therefore submitted to more invasive instrumental examinations. PMID- 15886625 TI - Trends in the diagnosis and the management of Meniere's disease: results of a survey. AB - OBJECTIVE: To determine the practices of the American Neurotology Society (ANS) membership in the evaluation and treatment of the Meniere's patient. STUDY DESIGN: Prospective. INTERVENTION: Questionnaire. MAIN OUTCOME MEASURE: Respondents' response to questions pertaining to the diagnostic and therapeutic practices in the management of Meniere's disease. RESULTS: Three hundred members of ANS were mailed a 15-item questionnaire. Two hundred three responded, for a 67.7% response rate. For the diagnosis of Meniere's disease, 1 in 3 practitioners relied solely on history, physical exam, and audiometry, whereas 2 in 3 relied in part on adjunctive tests, such as electrocochleography (ECOG) and electronystagmography (ENG). Two in 3 practitioners pursued retrocochlear studies on initial evaluation, with the overwhelming majority using MRI. In treating Meniere's disease, conservative medical management was preferred. For medically recalcitrant Meniere's disease, endolymphatic sac surgery (ESS) was the most commonly employed initial intervention (50%), followed by transtympanic gentamicin (38%). Currently, <10% routinely recommend the Meniett device. Eighty three percent include ESS as a therapeutic option for medically recalcitrant Meniere's disease. The vast majority continue to perform surgical labyrinthectomies and vestibular nerve sections for Meniere's disease. CONCLUSIONS: Meniere's disease continues to pose a difficult diagnostic and therapeutic problem, resulting in heterogeneous approaches to both evaluation and treatment. Despite the 1995 American Academy of Otolaryngology guidelines in the diagnosis of Meniere's disease, most clinicians rely in part on ENG or ECOG in diagnosing Meniere's disease. Furthermore, despite the passing of 20 years since the publications claiming a purely placebo effect, ESS is the most commonly employed initial surgical treatment for Meniere's disease. PMID- 15886626 TI - Use of Mimix hydroxyapatite bone cement for difficult ossicular reconstruction. AB - OBJECTIVE: To describe the advantages of using Mimix hydroxyapatite (HA) bone cement in reconstructing a variety of ossicular chain abnormalities. STUDY DESIGN AND SETTING: Case series at a tertiary medical center. RESULTS: Twenty-five cases of HA reconstruction are included in this series (ages 23-74; mean, 47 years). The examples presented include (1) HA as the sole reconstructive material for incus erosion, (2) HA for securing a total or partial ossicular replacement prosthesis, (3) incus augmentation after crimping for revision stapedotomy with incus erosion, (4) HA in primary stapedotomy to fix the crimped prosthesis to an intact incus, and (5) other unique situations. Preoperative and postoperative audiograms were evaluated for 4-tone pure tone average (PTA), speech reception thresholds, word recognition scores, and air-bone gaps (AB gaps). Mean follow-up was 11 months (range 2 to 22 months). The mean PTA improved from 57 dB to 37 dB, whereas the mean AB gaps decreased from 33 dB to 16 dB. There were no cases of infection or extrusion. CONCLUSIONS: Hydroxyapatite bone cement is an excellent adjunct or alternative to ossiculoplasty with preformed prostheses. Easily malleable, rapidly setting, and rapidly hardening, Mimix is particularly well suited for middle ear work. SIGNIFICANCE: Definitive fixation with bone cements during difficult ossicular chain reconstruction may ensure a more enduring successful outcome. PMID- 15886627 TI - Cochlear implantation in patients with osteogenesis imperfecta. AB - OBJECTIVE: Hearing loss has been shown to occur in 42% to 58% of patients with osteogenesis imperfecta (OI), with deafness arising in 25% to 60% of the patients. Implantation in patients with OI is relatively rare, with only 4 prior single case reports published in the English-language literature. The goal of this study was to evaluate the feasibility and functional outcome of cochlear implantation in 2 patients with OI tarda type I with profound sensorineural hearing loss. STUDY DESIGN: Case series. SETTING: The implantations were performed in a tertiary academic referral center (Johns Hopkins University). RESULTS: Though promontory vascularity was encountered, full insertion of a normal cochlear implant array could be achieved in both cases. One-year postimplant scores demonstrated 20 to 40 dB hearing thresholds, Consonant-Nucleus Consonant Test word scores of 54% and 70%, Consonant-Nucleus-Consonant Test phoneme scores of 75% and 83%, Hearing in Noise Test scores of 76% and 99%, and Central Institute of the Deaf Sentence Score sentence scores of 99% and 100%, for patients 1 and 2, respectively. CONCLUSIONS: Cochlear implantation in patients with OI is not only technically possible but the results are similar to implant outcomes for patients with sensorineural hearing loss from a variety of other causes. EBM RATING: C. PMID- 15886628 TI - Benign positional vertigo after cochlear implantation. AB - OBJECTIVE: To identify patients who underwent cochlear implantation (CI) and who subsequently developed benign positional vertigo (BPV) after the procedure and to identify any contributing factors. STUDY DESIGN AND SETTING: Academic tertiary referral center. Cochlear implant recipients' medical records were retrospectively reviewed to identify patients with both vertigo and, more specifically, BPV. Preoperative, intraoperative, and postoperative factors were studied vis-a-vis the development of BPV. RESULTS: BPV was newly diagnosed in 12 patients after CI. The etiology of hearing loss included presbycusis (16.6%), autoimmune inner ear disease (16.6%), congenital hearing loss (41.6%), Meniere's disease (8.3%), prematurity (8.3%), and idiopathic factors (8.3%). The onset of BPV varied after the procedure (mean +/- SD, 292 +/- 309 days). BPV symptoms did not affect implant performance. All patients were treated for BPV by Epley's maneuver and vestibular exercises. Symptoms disappeared in 11 patients and persisted in 1. CONCLUSIONS: BPV is an uncommon development after CI, although it occurs more frequently than in the general population. Two theories are proposed: the introduction of bone dust into the labyrinth and the dislodging of otoconia during surgery. The diagnosis, treatment, and prognosis of BPV after CI do not differ from those for non-CI-associated BPV. SIGNIFICANCE: Dizziness after CI usually develops as a result of vestibular hypofunction. BPV, which is a hyperfunctioning form of vestibular dysfunction, should be recognized as a possible sequelae of CI. PMID- 15886629 TI - Reliability of cochlear implants. AB - BACKGROUND: The long-term reliability of cochlear implants over time is an important issue for patients and cochlear implant teams. The calculation of cumulative survival rates including all hard failures of cochlear implants is suitable to report objectively about cochlear implant reliability. METHODS: This is a report of 192 cochlear implants from different manufacturers in adults (n = 58) and children (n = 134). RESULTS: The overall cumulative implant survival rate was 91.7% for a period of 11 years. The main reasons for hard failures were design errors of the products and direct or indirect trauma to the cochlea implant site (especially in children) with consecutive breaks of the implant body or electrodes. CONCLUSIONS: To improve our knowledge about reliability of cochlear implants more studies on cumulative long time survival of cochlear implants are needed, where functional failures and complications for whatever reason (design, mechanical, electronic, medical) are included. Cochlear implant reliability data should be considered during the choice of an implant for each individual patient. PMID- 15886630 TI - The "pull-back" technique for Nucleus 24 perimodiolar electrode insertion. AB - OBJECTIVE: To observe the influence of electrode pull-back after cochlear implant insertion of Nucleus 24 perimodiolar electrodes. STUDY DESIGN: In a prospective intraoperative study, we analyzed the impedances, neural response telemetry responses, and the spread of excitation after cochlear implant electrode insertion and compared these data to those obtained after a subsequent, controlled pull-back of the electrode. Postoperative depth of electrode insertion was controlled by x-ray. SETTING: Tertiary referral center. SUBJECTS: Six patients (4 male, 2 female; 18 to 69 years) were implanted with a Nucleus 24 (RCA) cochlear implant with a perimodiolar electrode. RESULTS: After a controlled pull-back, a significant decrease of the spread of excitation at the stimuli electrodes 5, 10, 15, and a nonsignificant decrease at stimuli electrode 20 compared to the recordings after the primary normal insertion procedure was found. The mean electric compound action potential amplitude was increased with an apical-to-basal tendency. Impedances remained unchanged by the pull-back. Mean insertion depth at the postoperative x-ray control was 372 degrees (+/- 10.2). CONCLUSION: Controlled cochlear implant electrode pull-back is a novel technique that optimizes objective intraoperative electrophysiological recordings in patients implanted with a Nucleus 24 perimodiolar cochlear implant by a greater approximation of the electrode to the modiolus. PMID- 15886631 TI - Tumor-dendritic cell fusion as a basis for cancer immunotherapy. AB - OBJECTIVE: To establish the basis for use of allogeneic dendritic-tumor fusion cells. STUDY DESIGN: Fusion cells were created by electrofusion. We used 2 allogeneic murine tumor lines (D5 and 4T1) that were virally transduced to express the antigen (beta-galactosidase) as a surrogate tumor marker. RESULTS: Cross-immunization was achieved with irradiated allogenic tumor cells. Successful electrofusion of dendritic cells and tumor cells was confirmed by using fluorescence-activated cell sorting and cytospin. Significant responses were shown in immunized mice against tumor challenge and established 3-day pulmonary metastasis with fusion cells. CONCLUSIONS: Allogeneic tumor sharing a common tumor antigen can immunize against syngeneic tumor challenge. Fusion cells showed successful immunization against tumor challenge and showed regression of 3-day established pulmonary metastasis. SIGNIFICANCE: These preclinical studies provide evidence that an allogenic tumor-dendritic cell fusion vaccine is a valid approach for head and neck cancer immunotherapy. PMID- 15886632 TI - Evaluation of tumor oxygenation by color duplex sonography: a new approach. AB - OBJECTIVE: Description of a new noninvasive method for the evaluation of tissue oxygenation in head and neck cancer. STUDY DESIGN AND SETTING: Prospective nonrandomized controlled study in an academic medical center on 20 patients with neck metastases of head and neck cancer. Metastases were investigated using color duplex sonography and pO2 histography. The vascularization in sonography was quantitatively evaluated by color pixel density and compared to the pO2 values of the same nodes. RESULTS: The correlation between vascularization and flow velocity was 0.71. For the mean/median pO2 -values and for the pO2 readings <10.0 mmHg correlations were r = 0.65 / 0.76 and 0.71. CONCLUSION: This sonographic method allows a safe and reliable evaluation of oxygenation in metastases of head and neck cancer. SIGNIFICANCE: The new approach is an alternative to pO2 histography and may play a future role in the planning of radiotherapy in the neck. PMID- 15886633 TI - Tracheal wall for the reconstruction of extended hemilaryngectomy in the rabbit model. AB - OBJECTIVE: To evaluate reconstruction of the surgical wound of extended hemilaryngectomy using the tracheal wall. STUDY DESIGN AND SETTING: Prospective, nonrandomized experimental study in a rabbit model in which cricothyroid hemilaryngectomy was performed in 10 rabbits. The proximal trachea was tailored to cover the surgical wound and sutured to it. The ipsilateral lobe of the thyroid gland and the medial thyroid vascular pedicle were conserved so as to assure vascular supply to the tracheal flap. After 4 weeks the animals were sacrificed and pathology analysis conducted. RESULTS: Eight of the 10 rabbits survived without respiratory or deglutitory alterations. Pathologic analysis showed well-healed suture lines together with normal mucosa and cartilage rings in the tracheal flap. CONCLUSIONS: The tracheal wall appears to be the ideal donor tissue for repair of the larynx. SIGNIFICANCE: This simple, 1-session surgical procedure demonstrates an effective reconstruction of an extended hemilaryngectomy wound. PMID- 15886634 TI - Virtual endoscopy of laryngeal carcinoma: is it useful? AB - OBJECTIVE: To compare virtual endoscopy (VE) with flexible endoscopy in patients with cancer of the larynx. STUDY DESIGN AND SETTINGS: This prospective study includes 24 patients with proven cancer of the larynx. Patients underwent spiral CT according to our standard protocol for upper airway imaging. This same set of axial scans was transferred to a dedicated workstation to obtain VE images. Results of VE were compared with the findings of flexible endoscopy. RESULTS: Quality of the examination was good in 96% of the patients. VE identified all exophytic lesions. Two small flat lesions could be observed as slightly enhanced plaques only on the axial scans. Subglottic extension was correctly demonstrated in all cases by associating VE to the axial scans. CONCLUSION: VE shows high sensitivity in the identification of exophytic lesions of the larynx and can establish relationships between cancer and nearby structures. It can be performed in the presence of severe stenosis and does not require sedation and additional scanning. On the other hand, VE show limits in the identification of flat lesions and does not allow biopsies and functional imaging to be performed. SIGNIFICANCE: VE is a useful tool for staging and presurgery treatment of cancer of the larynx. PMID- 15886635 TI - Oncologic and functional results of supracricoid partial laryngectomy with cricohyoidopexy. AB - OBJECTIVE: Supracricoid partial laryngectomy with cricohyoidopexy (SPL-CHP) is an alternative technique described for extensive tumors of the larynx that are beyond the limits of classical conservation partial laryngectomy and otherwise would be treated by total laryngectomy. STUDY DESIGN AND SETTING: Forty-six patients with carcinoma of the larynx underwent SPL-CHP between 1991 and 2003. The median age was 54 (range, 37 to 72). In 28 cases both arytenoids were spared; in 17 cases, 1 arytenoid was spared; and in 1 case, 2 arytenoids were resected. Bilateral elective neck dissections were performed in supraglottic carcinomas. In glottic carcinomas, neck dissection was performed in the presence of clinically positive lymph nodes. None of the patients were treated with postoperative radiation therapy. RESULTS: Forty-five patients were successfully decannulated; 1 patient with 2 arytenoids resected could not tolerate decannulation. The mean time for decannulation was 20 days (range, 9 to 60 days) when both arytenoids were spared with SPL-CHP, and 41 days (range, 13 to 150 days) for SPL-CHP when 1 arytenoid was spared. The average time for removal of the feeding tube was 21 days (range, 9 to 60 days) when both arytenoids were spared, and 40 days (range, 16 to 127 days) when 1 arytenoid was spared. The removal time of the feeding tube of the patient with 2 arytenoids resected was postoperative day 63. In 2 patients, aspiration pneumonia occurred as a result of swallowing impairment. In none of the patients temporary or permanent gastrostomy was needed. Two patients had local recurrence and were treated with a total laryngectomy; they are still alive. In 2 patients, secondary primary tumors were detected. The 3-year overall and cause-specific actuarial survival rates were 95.7 % and 87.5 % , respectively. CONCLUSIONS: Although prolonged hospitalization and delaying physiological functions can be termed as disadvantages of SPL-CHP, the operation is a reliable and oncologically valid procedure in selected cases of cancer of the larynx who would otherwise be operated by total laryngectomy. PMID- 15886636 TI - Four cases of simultaneous triple primary cancers of the hypopharynx, esophagus, and stomach. AB - OBJECTIVE: To study the efficacy of intensive chemotherapy for simultaneous triple primary cancers of the hypopharynx, esophagus, and stomach. STUDY DESIGN: Retrospective case study. METHODS: From April 2000 to March 2002, we treated 4 patients who had simultaneous triple primary cancers, including hypopharyngeal, esophageal, and gastric carcinomas. These 4 patients were designated as the objects for analysis, and the treatment modality for simultaneous multiple primary cancer cases was examined. RESULTS: In 3 of 4 patients, all 3 primary cancers could be controlled by intensive induction chemotherapy and concurrent chemoradiotherapy for hypopharyngeal cancer and by endoscopic mucosal resection or argon plasma coagulation for esophageal and gastric carcinomas. CONCLUSIONS: In the treatment modality for simultaneous multiple primary cancers, including head and neck cancer, it is important to select intensive systemic chemotherapy and decide the order for treating each primary lesion in consideration of each patient's curability and prognosis. PMID- 15886637 TI - Obstructive sleep apnea and head and neck neoplasms. AB - OBJECTIVE: To elaborate the relationship between obstructive sleep apnea (OSA) and head and neck neoplasms (HNN). STUDY DESIGN AND SETTING: A systematic review of the MEDLINE literature. RESULTS: Thirty of 34 articles indexed under OSA and HNN were about neoplasms that presented first as OSA. Four of the articles were about treatment of HNN causing OSA and gave incidences varying from 8% to 92%. Quality of life surveys confirm that patients with malignant HNN have sleep problems. Xerostomia may contribute to these sleep problems, and the role of salivary mucins deserves consideration. CONCLUSIONS: This review of the literature raises several new research questions about the clinical and biological relationships between OSA and HNN. PMID- 15886638 TI - A case of mucocutaneous leishmaniasis. PMID- 15886639 TI - Association between Kaposi's sarcoma and squamous cell carcinoma of the larynx. PMID- 15886640 TI - Simplifying video endoscopy: inexpensive analog and digital video endoscopy in otolaryngology. PMID- 15886641 TI - Hashimoto's thyroiditis presenting as an enlarging submandibular mass in a patient with a lingual thyroid. PMID- 15886642 TI - Cochlear implantation in a patient with bilateral temporal bone fractures. PMID- 15886643 TI - Inflammatory pseudotumor of the neck: a long-term result without surgical approach. PMID- 15886644 TI - Removal of a nasal leech: a safe and effective method. PMID- 15886645 TI - Frontal sinus tumor as the first sign of adenocarcinoma of the lung. PMID- 15886653 TI - Cost-effectiveness of different screening strategies for abdominal aortic aneurysm. AB - OBJECTIVE: The primary objective of this study was to develop a simulation model to assess the cost-effectiveness of different screening strategies for abdominal aortic aneurysms (AAAs) in men. METHODS: A systematic review of the literature was conducted for different screening strategies in terms of age (60, 65, or 70 years) and risk profiles (all men or specific high-risk groups) of the screened population, and rescreening after 5 or 10 years. These data were analyzed in a Markov simulation cohort model. RESULTS: The cost per life year gained for different screening strategies ranged from US 8,309 dollars to US 14,084 dollars and was estimated at US 10,474 dollars when 65-year-old men were screened once. Screening 60-year-old men was equally cost-effective, with the advantage of more life years gained. We demonstrated a trade-off between high prevalence of AAA and lower life expectancy, eliminating the expected benefits of screening high-risk groups such as smokers (US 10,695 dollars) or cardiovascular patients (US 10,392 dollars). Assuming general population utility resulted in a cost per quality adjusted life year (QALY) gained of US 13,900 dollars, whereas a hypothetical 5% reduction in utility among men with a screening-detected AAA raised the cost per QALY gained to US 75,100 dollars. CONCLUSION: This Markov model, which was based on a systematic review of the literature, supplied information on the estimated cost-effectiveness of different screening strategies. Screening men for AAA may be cost-effective in the long-term. Different screening strategies and quality-of life effects related to screening for AAA need to be evaluated in future clinical studies. PMID- 15886655 TI - Midterm outcome of endovascular repair of ruptured abdominal aortic aneurysms. AB - PURPOSE: We sought to analyze the clinical and morphologic outcomes of bifurcated stent grafts in patients with ruptured aortoiliac aneurysms at midterm follow-up. METHODS: Thirty-seven patients (4 women; mean age, 73 years; mean abdominal aortic aneurysm [AAA] diameter, 77 mm) underwent endovascular abdominal aneurysm repair between June 1997 and July 2003 for ruptured AAA. Devices inserted were as follows: Vanguard (Boston Scientific, Natick, Mass; n = 7), Excluder (W.L. Gore, Flagstaff, Ariz; n = 25), Talent (Medtronic Vascular, Santa Rosa, Calif; n = 2), and Zenith (Cook Inc, Bloomington, Ind; n = 3). Except for the adjunct postimplantation computed tomographic scanning, the imaging follow-up was the same as for nonruptured AAAs. RESULTS: The mean follow-up period was 24 months (range, 1-59) months. Thirty-day mortality was 10.8%. Three patients died during the follow-up of non-AAA-related causes. One patient was converted early for presumed renal overstenting. The late conversion rate was 9% because of stent graft migration (n = 2) or infection (n = 1). Freedom from endoleak was 57% +/- 8.5% and 48.8% +/- 9% at 2 and 4 years, respectively. Seventeen secondary interventions were performed during the follow-up period, 41% of these within 1 month of stent graft placement. Endoleaks, primary or secondary, were responsible for 58.8% of these interventions. The cumulative risk of a secondary intervention was 35.3% +/- 9% at 2 years and 44.6% +/- 11% at 3 years. Aneurysmal sac shrinkage was observed in 30.8% +/- 9.1% and sac enlargement was observed in 15.3% +/- 10.8% at 2 years. CONCLUSION: Endoluminal devices are able to convert the acute life-threatening situation of ruptured AAA to a controlled situation that results in good patient survival at midterm follow-up. PMID- 15886656 TI - Functional outcome after open repair of ruptured abdominal aortic aneurysm. AB - BACKGROUND: Outcome after operative repair of ruptured abdominal aortic aneurysm (AAA) has traditionally been assessed in terms of survival. This study examines the functional outcome of patients who survive operation. METHODS: Consecutive patients who survived open repair over an 18-month period were entered into a prospective case-control study. Age- and sex-matched controls were identified from patients undergoing elective AAA repair. The Short Form-36 health survey was administered to both groups of patients at 6 months after operation. Results were compared with the expected scores for an age- and sex-matched normal UK population. RESULTS: Fifty-seven patients underwent open repair of a ruptured AAA, and 30 survived; no patient was lost to follow-up. There were no significant differences in quality of life between patients who had an emergency repair and those who had an elective repair. Both of these groups had poorer health-related quality of life outcomes than the matched normal population. Surprisingly, compared with the normal population, patients after elective repair had poorer outcomes in more health domains than patients who survived emergency operation. CONCLUSIONS: Survivors of ruptured AAA repair have a good functional outcome within 6 months of operation. PMID- 15886657 TI - Determinants of carotid endarterectomy anatomic durability: effects of serum lipids and lipid-lowering drugs. AB - OBJECTIVES: Carotid endarterectomy (CEA) remains the gold standard for treatment of carotid stenosis. With inevitable comparisons of catheter-based therapy to all aspects of CEA, this study of a large contemporary series was undertaken to evaluate the determinants of anatomic durability of CEA. METHODS: During the interval (1989 through 1999), 2,127 primary, isolated CEAs with selective patching (50.2%) were performed in 1,853 patients (61.8% male, 36.1% symptomatic). End points included patient longevity and perioperative morbidity as well as evidence of CEA anatomic durability as defined by duplex evaluation: CEA restenosis (moderate, >50%, or greater recurrent stenosis), which included CEA anatomic failure (severe, >70%, restenosis/carotid occlusion). The incidence of CEA recurrent stenosis was temporally assessed early (<2 years) and late (>2 years) after operation. Clinical and surgical variables potentially associated with the study endpoints were analyzed by univariate and multivariate methods. RESULTS: The perioperative stroke and death rate was 1.4% and the 2-year and 10 year survival was 88.1% and 44.9%, respectively. Anatomic failure after CEA developed in 3.9% at 2 years and in 8.5% at 5 years; only 3.2% of CEA patients underwent reoperation during a mean follow-up of 73.4 months. Early (<2 years) analysis revealed 12.2% restenosis, whereas late (>2 years) results identified 9.8% progression of carotid stenosis and a 5.8% rate of anatomic failure. Multivariate analysis determined elevated creatinine (odds ratio [OR], 1.719, P < .001) and female gender (OR, 1.564; P < .02) correlated with early restenosis. Surgical patch closure and lipid-lowering drugs were protective for both early restenosis, with ORs of 0.543 (P < .0.001) and 0.601 (P < .007) and early anatomic failure ORs of 0.469 (P < .02) and 0.517 (P < .03), respectively. Although only elevated serum cholesterol (OR, 1.009; P < .03) correlated with late anatomic failure, only lipid-lowering drugs were protective for both late freedom from progression of disease (OR, 0.202; P < .0002) or late CEA anatomic failure (OR, 0.128; P < .0003). CONCLUSIONS: The association of female gender and elevated cholesterol with recurrent carotid stenosis is confirmed, elevated creatinine is introduced as a risk factor, and surgical patch repair is protective for early CEA recurrent carotid stenosis. The unique findings of the significant, beneficial effects of lipid-lowering drugs on both early and late CEA anatomic durability and patient survival indicate that such therapy should be instituted in most patients after CEA. PMID- 15886658 TI - Cardiac troponin I assessment and late cardiac complications after carotid stenting or endarterectomy. AB - PURPOSE: When compared with carotid endarterectomy (CEA), percutaneous carotid angioplasty with stent replacement (CAS) is a less invasive technique in the treatment of carotid stenosis. However, periprocedural hemodynamic instability still remains a challenge. This instability might lead to myocardial damage, which is now measured accurately by using cardiac troponin I (CTnI). METHODS: This study was designed to compare the periprocedural variation of CTnI in 150 consecutive patients scheduled to undergo CEA (n = 75) or CAS (n = 75). The levels of CTnI were measured until the third postoperative day in all patients. Short-term (1 month) and long-term (up to 5 years) postoperative cardiac outcome were assessed by means of chart review, regular follow-ups, and telephone calls. RESULTS: There was not any statistically significant difference between the 2 groups regarding the demographic characteristics and preprocedural medical status. The incidence of increase of CTnI (>0.5 ng/mL) was significantly higher in the CEA group (13%) compared with that in the CAS group (1%; P = .001). During the acute postprocedural period, the CAS group was significantly more prone to hypotension, requiring vasopressor therapy, whereas the CEA group had more hypertension, necessitating hypotensive medications (P < .001). At 5 years, the overall incidence of major cardiac complications (nonfatal myocardial infarction and death related to cardiac origin) was significantly more frequent in the CEA group (20% vs 5%, P < .01). CONCLUSION: The results of our study suggest that CAS yielded less myocardial damage in the short and long term when compared with CEA. Larger randomized multicenter trials with long-term outcomes are necessary to confirm our findings. PMID- 15886659 TI - Does carotid endarterectomy improve cognitive functioning? AB - BACKGROUND: Carotid endarterectomy (CEA) might improve cognitive functioning, but studies thus far have produced mixed results. The aim of the present study was to examine the effect of CEA on cognitive functions in a methodologically more strict design, first by testing the presumption of preoperative cognitive impairment and second through a better control for the possible influence of the nonspecific effects of practice and surgery. METHODS: Preoperative performance on a neuropsychologic test battery of 56 patients with severe occlusive disease of the carotid artery but without history of major stroke was compared with the performance of 46 healthy control subjects and 23 patients before endarterectomy of the superficial femoral artery (remote endarterectomy). The degree of cognitive change in the 2 patient groups was compared at 3 and 12 months postoperatively. We assessed mood to control for possible momentary affective influences on cognition. RESULTS: Before CEA, patients showed reduced functioning compared with that seen in healthy control subjects in terms of attention, verbal and visual memory, planning of motor behavior, psychomotor skills, and executive function. Performance of patients before remote endarterectomy was reduced as well. Improvements in several cognitive functions were observed after both types of surgical interventions and were attributed to psychologic relief from uncomplicated surgery and to practice. CONCLUSIONS: No specific restorative effect of CEA on cognitive functioning was observed. The preoperative impairment in several cognitive domains might be caused by factors that patients with various types of vascular disease might have in common, such as small-vessel disease or other undetected abnormalities within the brain. PMID- 15886661 TI - Pseudoaneurysm formation in a subset of patients with small intestinal submucosa biologic patches after carotid endarterectomy. AB - BACKGROUND: The carotid artery is frequently patched after carotid endarterectomy (CEA) to minimize the risks of early postoperative thrombosis and late recurrent stenosis. The small intestinal submucosa (SIS) patch is a biologic vascular patch derived from porcine small intestine. It is composed primarily of cell-free collagen and other extracellular matrix constituents that act as a scaffold for host cell deposition. METHODS: In May 2001, we began an investigational trial of SIS patches in 76 patients undergoing patch angioplasty of the carotid artery after CEA. RESULTS: No adverse events related to the patches were observed in the first 69 patients implanted with an SIS patch. However, in late 2002, seven patients were found to have asymptomatic pseudoaneurysms (PSA) by duplex imaging < or =10 weeks after their CEAs. The trial was immediately suspended. The PSAs were treated by surgical resection with vein grafting in two patients and placement of covered endoluminal stents in four patients. One patient is being followed as the PSA is small and has remained stable. Histopathologic examination of the SIS patch explanted from one of the surgically treated patients demonstrated the presence of actin-positive myofibroblasts or smooth muscle cells. Extensive mechanical testing of the SIS material from the two material lots associated with PSAs demonstrated thinner and more variable physical characteristics compared with control device lots. CONCLUSIONS: Biologic patches that undergo active remodeling in the carotid artery require greater thickness than was anticipated to decrease wall stress and suture hole elongation. Patches exceeding this minimum thickness will be required to ensure the safety of new SIS patch designs for vascular operations. PMID- 15886662 TI - Prospective study of carotid endarterectomy with modified polytetrafluoroethylene (ACUSEAL) patching: early and late results. AB - OBJECTIVE: Carotid endarterectomies (CEAs) with standard polytetrafluoroethylene (PTFE) patching have been shown to have results comparable with those of autogenous vein patching; however, prolonged bleeding through needle holes in PTFE is a commonly recognized problem. This is the first study of CEA using a new hemostatic modified PTFE patch (GORE-TEX) analyzing the early and late outcomes. METHODS: Two hundred consecutive CEAs were entered into this protocol. All patients had an immediate postoperative carotid duplex ultrasound scan that was repeated at 1 month and every 6 to 12 months thereafter. A Kaplan-Meier analysis was used to estimate the stroke-free survival and the risk of restenosis. The mean follow-up was 21 months (range, 1 to 48 months). RESULTS: The perioperative stroke rate was 1.5% (1% ipsilateral and 0.5% contralateral, two minor strokes and one major stroke) with no perioperative mortality or perioperative carotid thrombosis. The incidence of perioperative transient ischemic attacks was 3.5% (2.5% ipsilateral and 1% contralateral). The mean hemostasis time after completion of the patching was 3 minutes, in contrast to 14 minutes for conventional PTFE (in a previous study). The rates of freedom from ipsilateral strokes at 1, 2, 3, and 4 years were 99%, 99%, 99%, and 99%, respectively. The cumulative stroke-free survival rates at 1, 2, 3, and 4 years were 98%, 96%, 93%, and 93%, respectively. The rates of freedom from > or =70% restenosis at 1, 2, 3, and 4 years were 97%, 97%, 94%, and 94%, respectively. CONCLUSIONS: CEAs with a new modified PTFE patch are safe, have low perioperative stroke rates, are durable, and have an acceptable hemostasis time. PMID- 15886663 TI - Rapid foot and calf compression increases walking distance in patients with intermittent claudication: results of a randomized study. AB - OBJECTIVE: The aim of our pilot study was to determine the usefulness of rapid, high-pressure, intermittent pneumatic calf and foot compression (IPCFC) in patients with stable intermittent claudication, with reference to the end points of improvement in initial claudication distance (ICD) (distance at which patient feels pain or discomfort in the legs), and improvement in absolute claudication distance (ACD) (distance at which patient stops walking because the pain or discomfort becomes severe). METHODS: Thirty male patients presenting with stable, intermittent claudication (ACD between 50 and 150 meters on treadmill testing at 3.8 km/h, 10 degrees gradient) were recruited into this pilot study from a single center. Fifteen patients were randomized to treatment with IPCFC (applied for 1 hour twice daily in the sitting position) and were also advised to have daily exercise, and 15 patients served as controls, who were advised exercise alone. All patients received aspirin and had resting and postexercise ankle/brachial index (ABI) measured at enrollment along with ICD and ACD on treadmill testing (3.8 km/h, 10 degrees gradient). The mean age, baseline ICD, and ACD of the treatment and control groups were 70.4 +/- 7 years and 70.7 +/- 9 years, 55.8 +/- 15 meters and 68.4 +/- 17 meters, and 86.7 +/- 19 meters and 103.9 +/- 27 meters, respectively. Both groups were equally matched for risk factors, including smoking, type II diabetes mellitus, and hypercholesterolemia. IPCFC was applied. The study protocol included follow-up visits at 1, 2, 3, 4, 6, and 12 months with the ABI, ICD and ACD being measured at every visit. RESULTS: The percent change from baseline for ICD and ACD for each patient visit and the mean +/- standard deviation (SD), standard error (SE), and median were calculated for the control and treatment groups. The percent change from baseline measurements (mean +/- SD) for ICD and ACD in the control group at 4, 6, and 12 months were 2.2 +/- 18 and 2.3 +/- 18, 2.9 +/- 17 and 5.2 +/- 20, and 3.6 +/- 18 and 5.8 +/- 20, respectively. In contrast, the changes in ICD and ACD at 4, 6, and 12 months in the treatment group were 137.1 +/- 128 (P < .01) and 84.3 +/- 82 (P < .01), 140.6 +/- 127 (P < .01) and 96.4 +/- 106 (P = .01), and 150.8 +/- 124 (P <0.01) and 101.2 +/- 104 (P <0.01), respectively. Although the ABI showed a slight increase in the treatment group, these differences were not statistically significant. CONCLUSIONS: The results of this pilot study show that IPCFC improves walking distance in patients with stable intermittent claudication. A significant increase in ICD and ACD was seen at 4 and 6 months of treatment, respectively, and the improvement was sustained at 1 year. The combination of IPCFC with other treatment such as risk-factor modification and daily exercise may prove useful in patients with peripheral arterial occlusive disease. It may be a useful first line of therapy in patients with disabling claudication who are unfit for major reconstructive surgery. Improved walking on long-term follow-up and experience from different centers may establish a role for this treatment modality in the future. PMID- 15886664 TI - Skeletal muscle phenotype is associated with exercise tolerance in patients with peripheral arterial disease. AB - OBJECTIVE: To better understand the association between skeletal muscle and exercise intolerance in peripheral arterial disease (PAD), we assessed treadmill walking performance and gastrocnemius muscle phenotype in healthy control subjects and in patients with PAD. We hypothesized that gastrocnemius muscle characteristics would be altered in PAD compared with control subjects and that exercise tolerance in patients PAD would be related to muscle phenotype. METHODS: Sixteen patients with PAD and intermittent claudication and 13 healthy controls of the same age participated. Each subject completed a graded treadmill-walking test and underwent a resting muscle biopsy. Muscle biopsy samples were obtained from the medial gastrocnemius muscle of the most ischemic limb in PAD and a limb chosen at random in controls. Samples were analyzed for fiber type and cross sectional area, capillary-to-fiber ratio, the number of capillaries in contact with each fiber type, and the optical density of glycogen within each fiber by using histochemical procedures. Total muscle glycogen content was determined biochemically. RESULTS: Exercise capacity measured on the incremental walking test in the PAD group was only 30% to 40% of that observed in controls. The PAD group had a lower proportion of type I muscle fibers (P < .05), fewer capillaries per muscle fiber (P < .05), and tended to have smaller fiber areas (P = .08). The relative area of type I fibers, the capillary-to-fiber ratio, capillary contacts with type I and IIa fibers, and the optical density of glycogen in type I fibers were all positively correlated with exercise tolerance in the PAD group (P < .05) but not controls. CONCLUSIONS: These data suggest that muscle phenotype is altered in PAD and that such alterations are associated with the exercise intolerance in these patients. In light of these findings, therapies such as resistance training or electrical stimulation that target skeletal muscle in PAD may prove beneficial, and further investigation of such therapies is warranted. PMID- 15886665 TI - Factor V Leiden, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T mutations are not associated with chronic limb ischemia: the Linz Peripheral Arterial Disease (LIPAD) study. AB - OBJECTIVE: Factor V G1691A (Leiden), prothrombin G20210A, and methylenetetrahydrofolate reductase (MTHFR) C677T mutations are considered risk factors for venous thromboembolism. It remains to be characterized whether the presence of these relatively common mutations poses a risk for peripheral arterial disease (PAD). Therefore, we intended to test, by conducting a case control study, the hypothesis that PAD was associated with an increased prevalence of factor V G1691A, prothrombin G20210A, and MTHFR C677T mutations. METHODS: The study comprised 433 patients admitted for inpatient diagnostics and treatment of PAD in patients with chronic limb ischemia. Patients with acute ischemia or malignancy were excluded. A total of 433 control subjects matched to the patients with PAD in a 1:1 design by sex, age (+/-2 years), and diabetes mellitus status were recruited. Factor V G1691A, prothrombin G20210A, and MTHFR C677T genotypes were assessed by polymerase chain reaction. RESULTS: For the factor V G1691A polymorphism, the genotype frequencies in PAD patients were 92.8% GG (normal homozygotes = wild type) and 7.2% GA (mutant heterozygotes), and in control subjects they were 94.0% GG and 6.0% GA (chi 2 test; P = .493). The distribution of the prothrombin G20210A genotypes was 96.3% GG (normal homozygotes = wild type) and 3.7% GA (mutant heterozygotes) in PAD patients and was 97.2% GG and 2.8% GA in control subjects (chi 2 test; P = .442). Genotype frequencies for the MTHFR C677T polymorphism were 47.8% CC (normal homozygotes = wild type), 43.4% CT (mutant heterozygotes), and 8.8% TT (mutant homozygotes) in PAD patients, compared with 47.1% CC, 44.1% CT, and 8.8% TT in control subjects (chi 2 test; P = .977). Accordingly, as determined by logistic regression analysis, no significant odds ratios for heterozygous or homozygous genotypes of the three polymorphisms could be observed. CONCLUSIONS: PAD was not associated with an increased prevalence of factor V G1691A, prothrombin G20210A, and MTHFR C677T mutations in the population studied. Thus, there is no indication that of one of these mutations may be a risk factor for chronic limb ischemia. However, the role of these mutations in acute limb ischemia remains to be clarified. PMID- 15886666 TI - Atherosclerotic risk factor control in patients with peripheral arterial disease. AB - BACKGROUND: The presence of peripheral arterial disease (PAD), even in the absence of overt coronary artery disease (CAD), confers the same relative risk of death from a cardiovascular cause as in patients with a previous cardiovascular event. Current guidelines recommend atherosclerotic risk factor-reduction strategies in PAD patients identical to those in patients with a recent coronary event. The purpose of this study was to determine the status of atherosclerotic risk factor control in patients with PAD. METHODS: We analyzed the records of patients treated at 2 regional clinics serving 92,940 individuals. Full examination, laboratory, and pharmacy data were available for all patients. Pharmacy data were analyzed to determine prescriptions for beta-blocker therapy, angiotensin-converting enzyme inhibitors, and lipid-lowering agents. Lipid control was assessed through fasting lipid data. Glycemic control in diabetics was evaluated by using hemoglobin A 1c levels. RESULTS: We administratively identified 2839 patients with a diagnosis of PAD. The exclusion of 1106 patients with a diagnosis of CAD or validated not to have PAD resulted in a cohort of 1733 patients. Of these, 33.1% (574/1733) were currently receiving beta-blockers, 28.9% (500/1733) were receiving an angiotensin-converting enzyme inhibitor, and 31.3% (543/1733) were receiving a statin. Most patients (92%; 1594/1733) had a recent blood pressure recorded. However, 56% (893/1594) had a systolic blood pressure of 130 mm Hg or higher, 45.5% (726/1594) had a diastolic blood pressure of 80 mm Hg or higher, and 13.6% (217/1594) had a diastolic blood pressure of 90 mm Hg or higher. Screening fasting lipid profiles were found in 62.6% (1085/1733) of patients, 56% (508/912) had a low-density lipoprotein of 100 mg/dL or higher, and 21% (187/912) had a value of more than 130 mg/dL. In patients with diabetes, a hemoglobin A 1c level of 7.0% or higher was found in 54.2% (198/365) of patients. CONCLUSIONS: Despite national consensus of PAD as a CAD equivalent, patients are currently undertreated with regard to atherosclerotic risk factor modification. Until broader recognition of this disease process exists, vascular surgeons must continue to champion medical as well as surgical treatments for these patients. PMID- 15886667 TI - Racial differences in primary and repeat lower extremity amputation: results from a multihospital study. AB - OBJECTIVE: African Americans have a much higher risk of major (above- or below knee) lower extremity amputation and a lower rate of limb-preserving vascular surgery or angioplasty than white patients. This article analyzes two potential pathways for racial disparities: primary amputation, defined as a major amputation performed without any prior attempt at revascularization, and repeat amputation, defined as a major amputation subsequent to a previous through-foot or major amputation. METHODS: Randomly selected medical records were reviewed for 248 African American, 30 Hispanic, and 235 white or other-race patients undergoing above- or below-knee amputation between 1995 and 2003 at three Chicago teaching hospitals. Chronic disease prevalence and severity, preadmission functional status, clinical presentation, and vascular history were used to test the risk-adjusted effect of race and ethnicity on rates of primary and repeat amputation. RESULTS: Controlling for demographic, functional, chronic disease, and clinical characteristics, African American patients were 1.7 times more likely to have undergone both primary (P = .01) and repeat (P = .03) amputation than white or other-race amputees. Race remained a significant independent risk factor even after controlling for the higher severity of illness, greater disability, and more complex presentation of African American amputees. CONCLUSIONS: Higher rates of primary and repeat amputation for African American patients at study hospitals, which all have significant vascular surgery capacity and an aggressive policy of limb salvage, suggest that these rates may be even higher at less well equipped institutions. Improving access to primary and preventive care for lower-income patients could reduce amputation rates among African Americans. PMID- 15886668 TI - The role of epidural spinal cord stimulation in the treatment of Buerger's disease. AB - PURPOSE: This clinical, retrospective study is evaluated the effect of epidural spinal cord stimulation (SCS) in the treatment of Buerger's disease. METHODS: The clinical criteria of Shionoya were used to diagnose 29 patients (22 men, 7 women; mean age 33.7 years) with Buerger's disease. The patients underwent SCS. Complete physical examination and vascular laboratory data were available and recorded for all patients. Questions regarding the improvement of symptoms, in lifestyle, and in physical activities were asked direct interview or by telephone during mean follow-up of 4 years. RESULTS: The regional perfusion index (RPI), the ratio between the foot and chest transcutaneous oxygen pressure at baseline (before SCS treatment) was 0.27 +/- 0.25. Three months after SCS implantation the RPI increased to 0.41 +/- 0.22. During the follow-up period, a sustained improvement in microcirculation was recorded: the RPI at 1-year follow-up was 0.49 +/- 0.34 and at 3-year follow-up was 0.52 +/- 0.21. The most pronounced improvement in the RPI values was found in the subgroup of 13 patients with trophic lesions. In this group, the RPI increased significantly from 0.17 +/- 0.21 to 0.4 +/- 0.18 (P < .023) after a mean follow-up of 5.7 years. Two patients underwent major amputation of the lower target limb. The limb survival rate was 93.1%. During the follow-up period of 4 years, 21 of the 29 patients continued to smoke, and only five patients stopped nicotine exposure. CONCLUSIONS: We recorded a significant benefit in the microcirculation, a good limb survival rate, and the absence of new trophic lesions. During the follow-up period, no severe complications related to the implanted devices occurred. Because of the diffuse, distal, segmental nature of the disease, SCS should be considered as an alternative treatment modality in patients with Buerger's disease. PMID- 15886669 TI - Effectiveness of an extracellular matrix graft (OASIS Wound Matrix) in the treatment of chronic leg ulcers: a randomized clinical trial. AB - BACKGROUND: Venous leg ulcers are a major cause of morbidity, economic loss, and decreased quality of life in affected patients. Recently, biomaterials derived from natural tissue sources have been used to stimulate wound closure. One such biomaterial obtained from porcine small-intestine submucosa (SIS) has shown promise as an effective treatment to manage full-thickness wounds. Our objective was to compare the effectiveness of SIS wound matrix with compression vs compression alone in healing chronic leg ulcers within 12 weeks. METHODS: This was a prospective, randomized, controlled multicenter trial. Patients were 120 patients with at least 1 chronic leg ulcer. Patients were randomly assigned to receive either weekly topical treatment of SIS plus compression therapy (n = 62) or compression therapy alone (n = 58). Ulcer size was determined at enrollment and weekly throughout the treatment. Healing was assessed weekly for up to 12 weeks. Recurrence after 6 months was recorded. The primary outcome measure was the proportion of ulcers healed in each group at 12 weeks. RESULTS: After 12 weeks of treatment, 55% of the wounds in the SIS group were healed, as compared with 34% in the standard-care group (P = .0196). None of the healed patients treated with SIS wound matrix and seen for the 6-month follow-up experienced ulcer recurrence. CONCLUSIONS: The SIS wound matrix, as an adjunct therapy, significantly improves healing of chronic leg ulcers over compression therapy alone. PMID- 15886670 TI - Comparison of cell-type-specific vs transmural aortic gene expression in experimental aneurysms. AB - OBJECTIVE: Abdominal aortic aneurysm (AAA) progression and disease resistance are related to mural cellularity; adventitial macrophages and neocapillaries predominate in larger, advanced aneurysms, whereas smaller AAAs have fewer macrophages and retain more medial smooth muscle cells (SMCs). Expression analysis of mRNA derived from the entire aorta may mask the role that specific cell types play in modulating disease progression. We used laser capture microdissection (LCM) to isolate SMC and macrophage-predominant mural cell populations for gene expression analysis in variable-flow AAA. METHODS: Rat AAAs were created via porcine pancreatic elastase (PPE) infusion. Aortic flow was increased via femoral arteriovenous fistula creation (HF-AAA) or reduced via unilateral iliac ligation (LF-AAA) in selected cohorts. SMC and macrophage predominant cell populations were isolated via LCM and analyzed for expression of pro-inflammatory transcription factors and chemokines, cytokines, and proteolytic enzymes via real-time polymerase chain reaction. RESULTS: Aortic PPE infusion precipitated endothelial cell (EC) denudation, SMC apoptosis, and elastic lamellar degeneration. Increased aortic flow (HF > NF > LF) stimulated restorative EC and SMC proliferation (45.8 +/- 6.6 > 30.5 +/- 2.1 > 21 +/- 3.6 and 212.2 +/- 9.8 > 136.5 +/- 8.9 > 110 +/- 13.5, respectively, for both cell types; P < .05) at 5 days after PPE infusion, while simultaneously reducing medial SMC apoptosis and transmural macrophage infiltration. Expression of nuclear factor kappa B (NF-kappab), granulocyte macrophage-colony stimulating factor (GM-CSF), macrophage migration inhibitory (MIF), heparin-binding EGF-like factor (HB-EGF) and inducible nitric oxide synthase (iNOS) varied between cell types and flow conditions at all time points examined. Gelatinolytic protease expression varied by cell type in response to flow loading (eg, increased in SMCs, decreased in macrophages), consistent with observed patterns of elastolysis and SMC proliferation reported in prior experiments. CONCLUSIONS: Flow differentially regulates cell-specific AAA gene expression. Whole-organ analysis of AAA tissue lysates obscures important cellular responses to inflammation and flow, and may explain previous seemingly contradictory observations regarding proteolysis and cell proliferation. Cell-type specific expression and functional analyses may substantially clarify the pathophysiology of AAA disease. CLINICAL RELEVANCE: Understanding aneurysmal aortic degeneration at the most fundamental level is a critical precursor to the development of next-generation therapies such as drug-eluting endografts and/or medical therapies to limit expansion of preclinical AAA in high-risk or elderly patients. Although animal modeling is necessary to gain insight into the early initiating events of AAA disease, the methods used in such analyses have critical bearing on the conclusions drawn regarding pathogenesis and potential therapeutic derivations. By analyzing cell type-specific gene expression rather than whole-organ tissue lysates, the precise roles of important mediators such as metalloproteinases can be placed in the appropriate context. Further refinement of these techniques may allow cell specific therapies to be applied at defined time points in disease progression with improved patient outcome and reduced procedural morbidity. PMID- 15886671 TI - Identification of a homocysteine receptor in the peripheral endothelium and its role in proliferation. AB - BACKGROUND: Homocysteine, a risk factor for atherosclerosis, increases intimal hyperplasia after carotid endarterectomy with associated smooth muscle cell proliferation and modulation of cytokines. The N-methyl-D-aspartate receptor (NMDAr), a glutamate-gated ion channel receptor, is associated with homocysteine induced cerebrovascular injury; however, the receptor has not been identified in peripheral vascular cells, nor has any interaction with homocysteine been clarified. Our objectives were first, to identify NMDAr in rat carotid artery and rat aorta endothelial cells (RAEC); and second, to determine whether homocysteine activates NMDAr in the endothelium. METHODS: NR1 and NR2A, two NMDAr subunits, were probed in rat carotid arteries by immunohistochemistry. RNA was isolated from RAECs, and expression of all NMDAr subunits (NR1, 2A, 2B, 2C, and 2D) were examined by RT-PCR and sequencing. For receptor protein expression, RAEC were incubated with different homocysteine concentrations and incubation times and also were treated with 50 microM homocysteine and/or preincubated with 50 microM dizocilpine MK-801, an NMDAr inhibitor. RESULTS: Both NR1 and NR2A were expressed in rat carotid arteries. All NMDAr subunits were expressed in the RAECs, and there was 92% to 100% similarity compared with rat NMDAr from the National Center for Biotechnology Information (NCBI) GenBank. Homocysteine upregulated NR1 expression and increased cell proliferation. RAEC pretreatment with MK-801 reduced homocysteine-mediated cell proliferation. CONCLUSION: This study is the first to show that NMDAr exists in the peripheral vasculature, and that homocysteine may act via NMDAr to increase intimal hyperplasia. CLINICAL RELEVANCE: Our objectives included the identification of a homocysteine receptor in the peripheral vasculature. The possible inhibition of a homocysteine receptor to prevent intimal hyperplasia rather than treat established stenosis would make a significant clinical impact. This will open further avenues of study in determining the role of homocysteine in the pathogenesis of intimal hyperplasia. PMID- 15886672 TI - Ginsenoside Rb1 blocks homocysteine-induced endothelial dysfunction in porcine coronary arteries. AB - OBJECTIVE: Homocysteine (Hcy) is an independent risk factor for atherosclerosis. This study investigates the effects of ginsenoside Rb1, a major constituent of ginseng, on Hcy-induced endothelial dysfunction and molecular changes in porcine coronary arteries. METHODS: The coronary arteries were harvested from pig hearts and cut into 5-mm ring segments, which were then divided into six groups, including control, Hcy alone (50 microM), low-dose (1 microM) or high-dose (10 microM) Rb1 alone, and Hcy plus low-dose or high-dose Rb1. After 24-hour incubation, the rings were analyzed for vasomotor function in response to thromboxane A2 analog U46619, bradykinin, and sodium nitroprusside (SNP), respectively. In addition, superoxide anion was assessed by lucigenin-enhanced chemiluminescence analysis. Endothelial nitric oxide synthase (eNOS) was studied using real-time polymerase chain reaction and Western blot. RESULTS: Endothelium dependent relaxation (bradykinin) was significantly reduced in rings treated with Hcy alone as compared with the control (49.80% vs 71.77%, n = 8, P < .05), whereas neither high-dose nor low-dose Rb1 alone affected the endothelium dependent relaxation. The low-dose Rb1-Hcy combined group had a partially improved endothelium-dependent relaxation (54.44%), whereas the high-dose Rb1-Hcy combined group showed a complete recovery of endothelium-dependent relaxation (72.89%). There was no substantial difference in maximal contraction induced by U46619 or endothelium-independent relaxation by SNP among all groups (P > .05). Furthermore, superoxide anion was markedly increased by 137% in the Hcy-treated group as compared with the control, but there were no statistically significant changes from the control in all other groups (P > .05). Lastly, eNOS mRNA and protein levels were substantially reduced in the Hcy-treated group, but not in the Rb1-Hcy combined groups. CONCLUSIONS: This is the first study to show that ginsenoside Rb1 can effectively block Hcy-induced endothelial dysfunction and superoxide anion production as well as eNOS downregulation in porcine coronary arteries. This study suggests that ginseng and its active constituents may have potential clinical applications in controlling Hcy-associated vascular injuries. CLINICAL RELEVANCE: Homocysteine (Hcy) is an independent risk factor for atherosclerosis and other vascular lesions. It causes endothelial dysfunction and oxidative stress. Ginseng compounds have effects of vasorelaxation and antioxidation. The purpose of this study was to determine the effect of ginsenoside Rb1, a major constituent of ginseng, on Hcy-induced endothelial dysfunction and molecular changes in porcine coronary arteries. Our results showed that ginsenoside Rb1 can effectively block Hcy-induced dysfunction of endothelium-dependent vasorelaxation as well as superoxide anion production and eNOS downregulation. This study suggests that ginseng compounds may have potential clinical applications in controlling Hcy-associated vascular diseases and other vascular lesions. PMID- 15886673 TI - Differential effects of orbital and laminar shear stress on endothelial cells. AB - OBJECTIVE: Laminar shear stress is atheroprotective for endothelial cells (ECs), whereas nonlaminar, disturbed, or oscillatory shear stress correlates with development of atherosclerosis and neointimal hyperplasia. The effects of orbital and laminar shear stress on EC morphology, proliferation, and apoptosis were compared. METHODS: ECs were exposed to orbital shear stress with an orbital shaker (210 rpm) or laminar shear stress (14 dyne/cm 2) with a parallel plate. Shear stress in the orbital shaker was measured with optical velocimetry. Cell proliferation was assessed with direct counting and proliferating cell nuclear antigen staining; apoptosis was assessed with transferase-mediated deoxyuridine triphosphate nick end labeling staining. Cell surface E-selectin and intercellular adhesion molecule expression were assessed with fluorescence activated cell sorting. Akt phosphorylation was assessed with Western blotting. RESULTS: Orbital shear stress increased EC proliferation by 29% and 3 [H]thymidine incorporation two-fold compared to 16% and 38% decreases, respectively, in ECs treated with laminar shear stress (P < .0001 and P = .03, analysis of variance). Cells in the periphery of the culture well aligned to the direction of shear stress similar to the shape change seen with laminar shear stress, whereas ECs in the center of the well appeared unaligned similar to ECs not exposed to shear stress. Shear stress at the bottom surface of the culture well was reduced in the center of the well (5 dyne/cm 2) compared to the periphery (11 dyne/cm 2); the Reynolds' number was 2066. ECs were seeded differentially in the center and periphery of the wells. ECs in the center of the well had increased proliferation, increased apoptosis, reduced Akt phosphorylation, increased intercelluar adhesion molecule expression, and reduced E-selectin down-regulation, compared with ECs in the periphery of the well. CONCLUSION: Although the orbital shaker does not apply uniform shear stress throughout the culture well, arterial magnitudes of shear stress are present in the periphery of the well. ECs cultured in the center of the well exposed to low magnitudes of orbital shear stress might be a model of the "activated" EC phenotype. CLINICAL RELEVANCE: The perfect in vitro model to study and assess treatments for atherosclerosis and neointimal hyperplasia does not exists. An extensive body of literature describing effects of laminar shear stress on endothelial cells has contributed to our understanding of the interactions between shear stress and blood vessels. Laminar shear stress is atheroprotective, whereas oscillatory or disturbed shear stress correlates with areas of atherosclerosis and neointimal hyperplasia in vivo. This study describes the orbital shear stress model, its effects on endothelial cell proliferation and apoptosis, and suggests that activation of the intracellular Akt pathway is associated with these differing effects of laminar and orbital shear stress on endothelial cells. PMID- 15886674 TI - Combining open and endovascular surgery for the treatment of infrarenal abdominal aortic aneurysm: a case report using a hybrid vascular graft. AB - Migration and endoleaks after endovascular exclusion of an infrarenal abdominal aortic aneurysm may lead to long-term failure of the stent graft. We report a successful case of a novel technique that combined open and endovascular surgery to address the issues of migration and endoleak in the repair of an abdominal aortic aneurysm. The hybrid graft, consisting of a proximal, conventional Dacron graft and two distal endoprosthesis limbs, was designed to reduce aortic cross clamp time in aortic procedures. This is the first reported clinical experience with this new hybrid vascular graft for the treatment of an abdominal aortic aneurysm. PMID- 15886675 TI - Totally laparoscopic explantation of migrated stent graft after endovascular aneurysm repair: a report of two cases. AB - This report describes the removal of two migrated stent grafts and the repair of abdominal aortic aneurysms by laparoscopic technique. In these two cases, endovascular treatment was not indicated because of device migration into the aneurysm and the presence of thrombus within the endografts. Operative times were 245 and 230 minutes, with aortic clamp times of 95 and 66 minutes. The patients were extubated immediately after the procedure, resumed a normal diet on postoperative day 2, and were discharged home on postoperative days 5 and 6. We believe these are the first reported cases of laparoscopic explantation of migrated aortic stent grafts in the literature. PMID- 15886676 TI - Selective use of electrolytic detachable and fibered coils to embolize a wide neck giant splenic artery pseudoaneurysm. AB - We report on a 55-year-old woman with recurrent chronic pancreatitis complicated by a 7-cm splenic artery pseudoaneurysm. A therapeutic endovascular treatment was chosen because of the patient's high surgical risk. Initial embolization with fibered coils succeeded in thrombosing the aneurysm lumen, but the aneurysm neck remained patent. Further embolization was achieved with two detachable Gugliemi Detachable Coils (GDCs) deployed across the aneurysm neck, successfully thrombosing the residual aneurysm. GDCs complement the more thrombogenic traditional fibered coils and should be used advantageously in critical locations to embolize difficult splenic artery pseudoaneurysms. PMID- 15886677 TI - Idiopathic renal artery and infrarenal aortic aneurysms in a 6-year-old child: case report and literature review. AB - Multiple arterial aneurysms are distinctly rare in the pediatric population. Most arterial aneurysms in children are secondary to infections, trauma, arteritides, collagen vascular diseases, and other causes. True idiopathic aneurysms are the least common and less than a dozen reports of multiple idiopathic aneurysms have been published. We present a case of an idiopathic, symptomatic renal artery aneurysm with fistulization to the renal vein and a concomitant abdominal aortic aneurysm in a 6-year-old boy. The diagnostic workup, surgical treatment, pathologic findings, and a review of the literature are presented. PMID- 15886678 TI - Stent-graft repair of a splenic artery aneurysm. PMID- 15886679 TI - Cutting balloon angioplasty of resistant renal artery stenosis caused by fibromuscular dysplasia. AB - Balloon angioplasty is an established intervention to treat renovascular hypertension. Conventional balloon angioplasty is usually effective in cases of renovascular hypertension caused by fibromuscular dysplasia. In the present report, we describe two cases of renovascular hypertension caused by fibromuscular dysplasia in which stenotic lesions were resistant to conventional balloon angioplasty but were successfully managed by a cutting balloon. After cutting balloon angioplasty, systemic blood pressure was normalized in both patients, without the use of antihypertensive agents. Angioplasty by using a cutting balloon is a new therapeutic choice against renal artery stenosis that is resistant to dilation by a conventional balloon. PMID- 15886680 TI - Totally laparoscopic aortic repair: a new device for direct transperitoneal approach. AB - On the basis of our experience with more than 71 cases of totally laparoscopic aortic surgery by the retrocolic approach, we have developed a new technique by a simple transperitoneal approach. The purpose of this report is to describe that technique and the novel laparoscopic bowel retractor used to ensure stable exposure of the aorta. PMID- 15886682 TI - Who should protect the public against bad doctors? PMID- 15886683 TI - Regarding "Aortobifemoral graft infection with Mycobacterium tuberculosis: Treatment with abscess drainage, debridement, and long-term administration of antibiotic agents". PMID- 15886685 TI - Regarding "No evidence of a gender difference in carotid blood velocities across stenoses". PMID- 15886686 TI - Regarding "Ultrasound findings after radiofrequency ablation of the great saphenous vein". PMID- 15886688 TI - Regarding "Deep vein thrombosis after radiofrequency ablation of greater saphenous vein: A word of caution". PMID- 15886693 TI - Viral metagenomics. AB - Viruses, most of which infect microorganisms, are the most abundant biological entities on the planet. Identifying and measuring the community dynamics of viruses in the environment is complicated because less than one percent of microbial hosts have been cultivated. Also, there is no single gene that is common to all viral genomes, so total uncultured viral diversity cannot be monitored using approaches analogous to ribosomal DNA profiling. Metagenomic analyses of uncultured viral communities circumvent these limitations and can provide insights into the composition and structure of environmental viral communities. PMID- 15886694 TI - Stable isotope probing - linking microbial identity to function. AB - Stable isotope probing (SIP) is a technique that is used to identify the microorganisms in environmental samples that use a particular growth substrate. The method relies on the incorporation of a substrate that is highly enriched in a stable isotope, such as (13)C, and the identification of active microorganisms by the selective recovery and analysis of isotope-enriched cellular components. DNA and rRNA are the most informative taxonomic biomarkers and (13)C-labelled molecules can be purified from unlabelled nucleic acid by density-gradient centrifugation. The future holds great promise for SIP, particularly when combined with other emerging technologies such as microarrays and metagenomics. PMID- 15886695 TI - Microbial community genomics in the ocean. AB - Marine microbial communities were among the first microbial communities to be studied using cultivation-independent genomic approaches. Ocean-going genomic studies are now providing a more comprehensive description of the organisms and processes that shape microbial community structure, function and dynamics in the sea. Through the lens of microbial community genomics, a more comprehensive view of uncultivated microbial species, gene and biochemical pathway distributions, and naturally occurring genomic variability is being brought into sharper focus. Besides providing new perspectives on oceanic microbial communities, these new studies are now poised to reveal the fundamental principles that drive microbial ecological and evolutionary processes. PMID- 15886696 TI - Type-II diabetes and pancreatic cancer: a meta-analysis of 36 studies. AB - Pancreatic cancer is the eighth major form of cancer-related death worldwide, causing 227 000 deaths annually. Type-II diabetes is widely considered to be associated with pancreatic cancer, but whether this represents a causal or consequential association is unclear. We conducted a meta-analysis to examine this association. A computer-based literature search from 1966 to 2005 yielded 17 case-control and 19 cohort or nested case-control studies with information on 9220 individuals with pancreatic cancer. The age and sex-adjusted odds ratio (OR) for pancreatic cancer associated with type-II diabetes was obtained from each study. The combined summary odds ratio was 1.82 (95% confidence interval (95% CI) 1.66-1.89), with evidence of heterogeneity across the studies (P=0.002 for case control and P=0.05 for cohort studies) that was explained, in part, by higher risks being reported by smaller studies and studies that reported before 2000. Individuals in whom diabetes had only recently been diagnosed (< 4 years) had a 50% greater risk of the malignancy compared with individuals who had diabetes for > or =5 years (OR 2.1 vs 1.5; P=0.005). These results support a modest causal association between type-II diabetes and pancreatic cancer. PMID- 15886697 TI - Guidelines for confidentiality and cancer registration. PMID- 15886698 TI - The polyAT, intronic IVS11-6 and Lys939Gln XPC polymorphisms are not associated with transitional cell carcinoma of the bladder. AB - Chemical carcinogens from cigarette smoking and occupational exposure are risk factors for bladder transitional cell carcinoma (TCC). The Xeroderma Pigmentosum Group C (XPC) gene is essential for repair of bulky adducts from carcinogens. The Xeroderma Pigmentosum Group C gene polymorphisms may alter DNA repair capacity (DRC), thus giving rise to genetic predisposition to bladder cancer. Recent studies have demonstrated linkage disequilibrium between three polymorphisms in the XPC gene (polyAT, IVS11-6 and Lys939Gln) and these have been shown to influence the DRC, as well as to be associated with bladder cancer risk. We analysed all three XPC polymorphisms in 547 bladder TCC patients and 579 cancer free controls to investigate the association between these polymorphisms and bladder cancer susceptibility, and we also attempted to assess gene-environmental interactions. We confirmed strong linkage disequilibrium among the polymorphisms (Lewontin's D' > 0.99). Using logistic regression adjusting for smoking, occupational and family history, neither the heterozygote nor the homozygote variants of these polymorphisms were associated with increased bladder cancer risk (adjusted odds ratio [95% confidence interval] for heterozygote 0.82 [0.63 1.07], 0.82 [0.63-1.08] and 0.83 [0.63-1.08] for PolyAT, IVS11-6 and Lys939Gln, respectively and homozygote variant, 0.98 [0.68-1.42], 0.99 [0.69-1.43] and 1.01 [0.70-1.46]). Moreover, we did not find any significant interaction between these XPC polymorphisms and environmental exposure to cigarette smoking and occupational carcinogens. PMID- 15886699 TI - The DNA mismatch repair gene hMSH2 is a potent coactivator of oestrogen receptor alpha. AB - The DNA mismatch repair gene is a key regulator in the elimination of base-base mismatches and insertion/deletion loops (IDLs). Human MutS homologue 2 (hMSH2), originally identified as a human homologue of the bacterial MutS, is a tumour suppressor gene frequently mutated in hereditary non-polyposis colorectal cancer. Hereditary non-polyposis colorectal cancer is characterised by the early onset of colorectal cancer and the development of extracolonic cancers such as endometrial, ovarian, and urological cancers. Oestrogen receptor (ER) alpha and beta are members of a nuclear receptor (NR) superfamily. Ligand-dependent transcription of ER is regulated by the p160 steroid receptor coactivator family, the thyroid hormone receptor-associated proteins/the vitamin D receptor interacting proteins (TRAP/DRIP) mediator complex, and the TATA box-binding protein (TBP)-free TBP associated factor complex (TFTC) type histone acetyltransferase complex. Here, we report the interaction between ER alpha/beta and hMSH2. Immunoprecipitation and glutathione-S-transferase pull-down assay revealed that ER alpha and hMSH2 interacted in a ligand-dependent manner, whereas ER beta and hMSH2 interacted in a ligand-independent manner. Oestrogen receptor alpha/beta bound to hMSH2 through the hMSH3/hMSH6 interaction domain of hMSH2. In a transient expression assay, hMSH2 potentiated the transactivation function of liganded ER alpha, but not that of ER beta. These results suggest that hMSH2 may play an important role as a putative coactivator in ER alpha dependent gene expression. PMID- 15886700 TI - Cancer incidence and mortality in patients with insulin-treated diabetes: a UK cohort study. AB - Raised risks of several cancers have been found in patients with type II diabetes, but there are few data on cancer risk in type I diabetes. We conducted a cohort study of 28 900 UK patients with insulin-treated diabetes followed for 520 517 person-years, and compared their cancer incidence and mortality with national expectations. To analyse by diabetes type, we examined risks separately in 23 834 patients diagnosed with diabetes under the age of 30 years, who will almost all have had type I diabetes, and 5066 patients diagnosed at ages 30-49 years, who probably mainly had type II. Relative risks of cancer overall were close to unity, but ovarian cancer risk was highly significantly raised in patients with diabetes diagnosed under age 30 years (standardised incidence ratio (SIR)=2.14; 95% confidence interval (CI) 1.22-3.48; standardised mortality ratio (SMR)=2.90; 95% CI 1.45-5.19), with greatest risks for those with diabetes diagnosed at ages 10-19 years. Risks of cancer at other major sites were not substantially raised for type I patients. The excesses of obesity- and alcohol related cancers in type II diabetes may be due to confounding rather than diabetes per se. PMID- 15886701 TI - A case-control study in Shanghai of fruit and vegetable intake and endometrial cancer. AB - In a population-based case-control study of 832 incident endometrial cancer cases and 846 frequency-matched controls among Chinese women in Shanghai, using a validated food-frequency questionnaire, dietary habits were estimated by in person interviews. Total vegetable consumption was inversely associated with endometrial cancer risk (highest quartile vs lowest: OR=0.69, 95% CI 0.50-0.96). The risk was reduced with increasing intake of dark green/dark yellow vegetables (trend test, P=0.02), fresh legumes (trend test, P<0.01), and allium vegetables (trend test, P=0.04). Fruit consumption was unrelated to risk. These results suggest that high consumption of certain vegetables may reduce the risk of endometrial cancer. PMID- 15886702 TI - Alcohol dehydrogenase 1B (ADH1B) genotype, alcohol consumption and breast cancer risk by age 50 years in a German case-control study. AB - In a population-based study of 613 cases and 1082 controls, alcohol dehydrogenase 1B (ADH1B) genotype was not an independent risk factor for breast cancer, although the possibility was raised that it modifies risk associated with high levels of alcohol consumption (OR 1.1, 95% confidence interval (CI) 0.8-1.6 for ADH1B*1/*1 genotype vs 0.2, 95% CI 0.1-1.0 for ADH1B*2 carriers). PMID- 15886703 TI - Urbanisation and incidence of acute lymphocytic leukaemia among United States children aged 0-4. AB - Acute lymphocytic leukaemia (ALL) incidence among children under 5 years of age was examined, utilising data from 24 United States cancer registries. County based incidence rates among white children were compared across four levels of urbanisation: large and small metropolitan counties, and adjacent and nonadjacent rural counties. In metropolitan areas, the incidence of ALL was lower among blacks (rate ratio (RR)=0.38, confidence interval (CI)=0.33-0.44) and among Asians/Pacific Islanders (RR=0.78, CI=0.63-0.97) than among whites. Among white children, the incidence of ALL decreased across the four strata of urbanisation, from 67 to 62 to 65 to 54 cases per million person-years at-risk (two-sided trend P=0.009), such that rates were significantly lower in the most remote rural counties than in the most populous metropolitan counties (RR=0.80, 95% CI=0.70 0.91). PMID- 15886704 TI - Enhanced photodynamic destruction of a transplantable fibrosarcoma using photochemical internalisation of gelonin. AB - Photochemical internalisation (PCI) is a technique for releasing biologically active macromolecules from endocytic vesicles by light activation of a photosensitiser localised in the same vesicles of targeted cells. This study investigated the PCI of the toxin gelonin as a way of enhancing the effect of photodynamic therapy (PDT) on a human malignant fibrous histiocytoma transplanted into nude mice using the photosensitiser disulphonated aluminium phthalocyanine (AlPcS(2a)). Pharmacokinetic studies after intraperitoneal administration showed that the serum level of AlPcS(2a) fitted a biexponential model (half-lives of 1.8 and 26.7 h). The tumour concentration was roughly constant up to 48 h, although fluorescence microscopy showed that the drug location was initially mainly vascular, but became intracellular by 48 h. To compare PDT with PCI, 48 h after intraperitoneal injection of 10 mg kg(-1) AlPcS(2a), and 6 h after direct intratumour injection of 50 microg gelonin (PCI) or a similar volume of phosphate buffered saline (PDT controls), tumour-bearing animals were exposed to red light (150 J cm(-2)). Complete response was observed for more than 100 days in 50% of the PCI tumours but only 10% of the PDT tumours (P<0.01). In tumours examined histologically 4 days after light delivery, the depth of necrosis was 3-4 mm after PDT, but 7 mm after PCI. The deeper effect after PCI demonstrates that the light fluence needed to kill tumour is less than with PDT. We conclude that PCI with gelonin can markedly enhance the effect of PDT on this type of tumour and may have a role clinically as an adjunct to surgery to control localised disease. PMID- 15886705 TI - Expression of Ksp-cadherin during kidney development and in renal cell carcinoma. AB - Cadherins are a large family of cell-cell adhesion molecules acting in a homotypic, homophilic manner that play an important role in the maintenance of tissue integrity. In the human kidney, several members of the cadherin family (including E- and N-cadherin, cadherin-6, -8 and -11) are expressed in a controlled spatiotemporal pattern. Cadherin-16, also called kidney-specific (Ksp ) cadherin, is exclusively expressed in epithelial cells of the adult kidney. In renal cell carcinomas (RCCs), which are considered to originate from epithelial kidney tubular cells, a complex pattern of cadherin expression can be observed, but Ksp-cadherin expression has not been analysed so far. In the present study, we show that the expression of Ksp-cadherin is completely abrogated in RCCs. Whereas Ksp-cadherin can be detected at later stages of tubulogenesis during human renal development and in the distal tubules of adult kidneys, no expression was found by immunohistochemistry or Western blot analysis in RCC tumour tissues and several RCC cell lines. However, despite the lack of protein expression, mRNA synthesis of Ksp-cadherin could be detected by reverse transcriptase-polymerase chain reaction analysis in all RCC tissues and most of the RCC cell lines studied, although at a reduced level. The loss of Ksp-cadherin protein was only observed in the malignant part of the tumour kidneys, whereas in the normal part of the affected kidneys Ksp-cadherin expression was clearly detected. These results indicate a downregulation of Ksp-cadherin in RCC and suggest a role for this cell adhesion molecule in tumour suppression. PMID- 15886706 TI - Phase I trial combining gemcitabine and treosulfan in advanced cutaneous and uveal melanoma patients. AB - Gemcitabine and treosulfan are DNA-damaging agents. Preclinical studies suggest that synergism exists when melanoma cells are exposed to both drugs concurrently. We conducted a phase I trial in advanced melanoma patients to determine the optimal dose of gemcitabine to be combined with treosulfan. Cohorts of three patients received increasing doses of gemcitabine, commencing at 0.5 g m(-2), followed by a fixed dose of 5.0 g m(-2) treosulfan on day one of a 21-day cycle. Patients alternately received a first cycle of single-agent gemcitabine or treosulfan before subsequent cycles of both drugs. Peripheral blood lymphocytes were collected in cycles 1 and 2 at various time points until 48 h post treatment. The single-cell gel electrophoresis (Comet) assay was used to measure chemotherapy-induced DNA damage. A total of 27 patients were enrolled, no objective responses were observed, but two uveal melanoma patients had minor responses. Dose-limiting myelosuppression was reached at 3.0 g m(-2) gemcitabine. DNA single-strand breaks were detected 4 h post-gemcitabine, repaired by 24 h. DNA interstrand crosslinks were detected 4 h post-treosulfan, fully removed by 48 h. Following combination chemotherapy, treosulfan-induced DNA crosslinks persisted, still being detectable 48 h post-treatment, supporting the hypothesis that gemcitabine potentiates treosulfan-induced cytotoxicity. The recommended regimen for further study is 2.5 g m(-2) gemcitabine combined with 5.0 g m(-2) treosulfan. PMID- 15886707 TI - Social and geographic disparities in access to reference care site for patients with colorectal cancer in France. AB - The aim of this study was to investigate the relationship between social and geographic characteristics and the type of care centre for initial colorectal surgery in France. Patients living far from a reference cancer site were less frequently treated in a reference cancer site than those who were living near a reference cancer site OR(a)=(0.50 (0.33-0.76)). As for topography and emergency presentation, place of residence (urban/rural), occupation and marital status were not associated with the type of the care centre. Improvements in diagnosis and treatment and of clinical practice guidelines are therefore crucial to ensure equality in health care in France. PMID- 15886708 TI - Plucked human hair as a tissue in which to assess pharmacodynamic end points during drug development studies. AB - We have demonstrated the feasibility of detecting and quantifying six cell-cycle related nuclear markers (Ki67, pRb, p27, phospho-p27 (phosphorylated p27), phospho-pRb (phosphorylated pRb), phospho-HH3 (phosphorylated histone H3)) in plucked human scalp and eyebrow hair. Estimates of the proportion of plucked hairs that are lost or damaged during processing plus the intra- and intersubject variability of each nuclear marker with these techniques are provided to inform sizing decisions for intervention studies with drugs potentially impacting on these markers in the future. PMID- 15886709 TI - Bioinformatics: computers or clinicians for complex disease risk assessment? PMID- 15886710 TI - The population history of the Croatian linguistic minority of Molise (southern Italy): a maternal view. AB - This study examines the mitochondrial DNA (mtDNA) diversity of the Croatian speaking minority of Molise and evaluates its potential genetic relatedness to the neighbouring Italian groups and the Croatian parental population. Intermatch, genetic distance, and admixture analyses highlighted the genetic similarity between the Croatians of Molise and the neighbouring Italian populations and demonstrated that the Croatian-Italian ethnic minority presents features lying between Croatians and Italians. This finding was confirmed by a phylogeographic approach, which revealed both the prevalence of Croatian and the penetrance of Italian maternal lineages in the Croatian community of Molise. These results suggest that there was no reproductive isolation between the two geographically proximate, yet culturally distinct populations living in Italy. The gene flow between the Croatian-Italians and the surrounding Italian populations indicate, therefore, that ethnic consciousness has not created reproductive barriers and that the Croatian-speaking minority of Molise does not represent a reproductively isolated entity. PMID- 15886711 TI - Mitochondrial DNA and ACTN3 genotypes in Finnish elite endurance and sprint athletes. AB - Differences in ACTN3 (alpha-actinin 3) genotypes have been reported among endurance and power athletes. Elite athletic performance in endurance sports should also depend on mitochondrial oxidative phosphorylation (OXPHOS) that produces ATP for muscle metabolism. We determined mitochondrial DNA (mtDNA) and ACTN3 genotypes in Finnish elite endurance (n = 52) and sprint (n = 89) athletes, and found that the frequencies of mtDNA haplogroups differed significantly between the two groups. Most notably, none of the endurance athletes belonged to haplogroup K or subhaplogroup J2, both of which have previously been associated with longevity. The frequency of ACTN3 XX genotype was higher and that of RR was lower among Finnish endurance athletes, and, in addition, none of the top Finnish sprinters had the XX genotype. Lack of mtDNA haplogroup K and subhaplogroup J2 among elite endurance athletes suggests that these haplogroups are 'uncoupling genomes'. Such genomes should not be beneficial to endurance-type athletic performance but should be beneficial to longevity, since uncoupling of OXPHOS reduces the production of ATP, reduces the release of reactive oxygen species and generates heat. PMID- 15886712 TI - Hutterite brothers both affected with two forms of limb girdle muscular dystrophy: LGMD2H and LGMD2I. AB - Limb girdle muscular dystrophy (LGMD) is very common in the Hutterite population of the North American Prairies. We have recently reported the homozygous c.1459G>A mutation in TRIM32 associated with LGMD2H. We have also identified Hutterite patients with LGMD2I, homozygous for the common c.826C>A mutation in FKRP. To date, all Hutterites with LGMD have been shown to be homozygous for either the TRIM32 or FKRP mutation. We now report a Hutterite family in which both parents and five sons were all found to be homozygous for the TRIM32 mutation. The father had slowly progressive proximal muscle weakness, whereas three sons and their mother, all currently asymptomatic, had normal physical examinations. The remaining two sons (7 and 10 years old), presented with mild decrease in stamina, had normal neuromuscular examinations and were found to be homozygous for the FKRP mutation in addition to the TRIM32 mutation. These two boys do not differ in age at or mode of presentation, physical findings, or serum CK levels compared to age-matched individuals affected with LGMD2I alone. This suggests that the effects of these two mutations are not acting synergistically at this time. It remains to be seen whether there will be signs of interaction between these two mutations as the patients get older. PMID- 15886713 TI - Short tandem repeats haplotyping of the HLA region in preimplantation HLA matching. AB - Recently, preimplantation genetic diagnosis (PGD) has been considered for several indications beyond its original purpose, not only to test embryos for genetic disease but also to select embryos for a nondisease trait, such as specific human leukocyte antigen (HLA) genotypes, related to immune compatibility with an existing affected child in need of a haematopoetic stem cell (HSC) transplant. We have optimized an indirect single-cell HLA typing protocol based on a multiplex fluorescent polymerase chain reaction (PCR) of short tandem repeat (STR) markers scattered throughout the HLA complex. The assay was clinically applied in 60 cycles from 45 couples. A conclusive HLA-matching diagnosis was achieved in 483/530 (91.1%) of the embryos tested. In total, 74 (15.3%) embryos revealed an HLA match with the affected siblings, 55 (11.4%) of which resulted unaffected and 46 (9.5%) have been transferred to the patients. Nine pregnancies were achieved, five healthy HLA-matched children have already been delivered and cord blood HSCs, were transplanted to three affected siblings, resulting in a successful haematopoietic reconstruction. PMID- 15886714 TI - Safety in numbers. PMID- 15886715 TI - RAR-related orphan receptor A isoform 1 (RORa1) is disrupted by a balanced translocation t(4;15)(q22.3;q21.3) associated with severe obesity. AB - We have identified a family comprising a mother and two children with idiopathic and profound obesity body mass index (BMI) 41-49 kg/m(2). The three family members carry a balanced reciprocal chromosome translocation t(4;15). We present here the clinical features of the affected individuals as well as the physical mapping and cloning of the chromosomal breakpoints. A detailed characterisation of the chromosomal breakpoints at chromosomes 4 and 15 revealed that the translocation is almost perfectly balanced with a very short insertion/deletion. The chromosome 15 breakpoint is positioned in intron 1 of the RAR-related orphan receptor A isoform 1 (RORa1) and the chromosome 4 breakpoint is positioned 133 kb telomeric to the transcriptional start of the unc-5 homolog B (UNC5C) and 154 kb centromeric of the transcriptional start of the pyruvate dehydrogenase (lipoamide) alpha 2 (PDHA2). The rearrangement creates a fusion gene, which includes the RORa1 exon 1 and UNC5C that is expressed in frame in adipocytes from the affected patients. We also show that this transcript is translated into a protein. From previous reports, it is shown that RORa1 is implicated in the regulation of adipogenesis and lipoprotein metabolism. We hypothesise that the obesity in this family is caused by (i) haploinsufficiency for RORa1 or, (ii) a gain of function mechanism mediated by the RORa1-UNC5C fusion gene. PMID- 15886716 TI - Cost/variance optimization for human exposure assessment studies. AB - The National Human Exposure Assessment Survey (NHEXAS) field study in EPA Region V (one of three NHEXAS field studies) provides extensive exposure data on a representative sample of 249 residents of the Great Lakes states. Concentration data were obtained for both metals and volatile organic compounds (VOCs) from multiple environmental media and from human biomarkers. A variance model for the logarithms of concentration measurements is used to define intraclass correlations between observations within primary sampling units (PSUs) (nominally counties) and within secondary sampling units (SSUs) (nominally Census blocks). A model for the total cost of the study is developed in terms of fixed costs and variable costs per PSU, SSU, and participant. Intraclass correlations are estimated for media and analytes with sufficient sample sizes. We demonstrate how the intraclass correlations and variable cost components can be used to determine the sample allocation that minimizes cost while achieving pre-specified precision constraints for future studies that monitor environmental concentrations and human exposures for metals and VOCs. PMID- 15886717 TI - The selective 5-HT2A receptor antagonist M100907 enhances antidepressant-like behavioral effects of the SSRI fluoxetine. AB - The addition of low doses of atypical antipsychotic drugs, which saturate 5 HT(2A) receptors, enhances the therapeutic effect of selective serotonin (5 hydroxytryptamine; 5-HT) reuptake inhibitors (SSRIs) in patients with major depression as well as treatment-refractory obsessive-compulsive disorder. The purpose of the present studies was to test the effects of combined treatment with a low dose of a highly selective 5-HT(2A) receptor antagonist (M100907; formerly MDL 100,907) and low doses of a SSRI using a behavioral screen in rodents (the differential-reinforcement-of low rate 72-s schedule of reinforcement; DRL 72-s) which previously has been shown to be sensitive both to 5-HT(2) antagonists and SSRIs. M100907 has a approximately 100-fold or greater selectivity at 5-HT(2A) receptors vs other 5-HT receptor subtypes, and would not be expected to appreciably occupy non-5-HT(2A) receptors at doses below 100 microg/kg. M100907 increased the reinforcement rate, decreased the response rate, and shifted the inter-response time distributions to the right in a pattern characteristic of antidepressant drugs. In addition, a positive synergistic interaction occurred when testing low doses of the 5-HT(2A) receptor antagonist (6.25-12.5 microg/kg) with clinically relevant doses of the SSRI fluoxetine (2.5-5 mg/kg), which both exerted minimal antidepressant-like effects by themselves. In vivo microdialysis study revealed that a low dose of M100907 (12.5 microg/kg) did not elevate extracellular 5-HT levels in the prefrontal cortex over those observed with fluoxetine alone (5 mg/kg). These results will be discussed in the context that the combined blockade of 5-HT(2A) receptors and serotonin transporters (SERT) may result in greater efficacy in treating neuropsychiatric syndromes than blocking either site alone. PMID- 15886718 TI - Reinstatement of morphine-conditioned reward is blocked by memantine. AB - Protection of abstinent individuals from relapse is the main goal of drug dependence treatment. Relapse is frequently precipitated by exposure to small doses of the drug of abuse or exposure to the environment that was previously associated with the drug. Mice exposed to morphine (10 mg/kg) in a unique test box environment display a conditioned place preference for this environment. Such preference can be extinguished by subsequent pairing of physiological saline administration with the same environment. Once extinguished, the original place preference can be reinstated after a priming dose (1-2.5 mg/kg) of morphine is given. However, mice treated with 7.5 (but not 3.75) mg/kg of memantine (the glutamate/NMDA receptor antagonist) during the extinction phase were insensitive to morphine's ability to reinstate the place preference 2 days after extinction conditionings. Effect of memantine was also observed when priming dose of morphine was given 21 days after extinction conditionings. In contrast, morphine's ability to reinstate conditioned response was not affected by treatment with 10 mg/kg of chlordiazepoxide, 0.5 mg/kg of LSD-25, or 1 mg/kg of morphine given during extinction conditionings. A separate experiment demonstrated that memantine (7.5 mg/kg) treatment did not affect learning. We show for the first time that memantine treatment during extinction conditionings may abolish the ability of drug-related cues to evoke reinstatement, suggesting that this NMDA receptor antagonist can be useful in preventing relapse in opioid dependent individuals. PMID- 15886719 TI - Cortical glutamatergic markers in schizophrenia. AB - Post-mortem studies have yet to produce consistent findings on cortical glutamatergic markers in schizophrenia; therefore, it is not possible to fully understand the role of abnormal glutamatergic function in the pathology of the disorder. To better understand the changes in cortical glutamatergic markers in schizophrenia, we measured the binding of radioligands to the ionotropic glutamate receptors (N-methyl D-aspartate, [3H]CGP39653, [3H]MK-801), amino-3 hydroxy-5-methyl-4-isoxazole ([3H]AMPA), kainate ([3H]kainate), and the high affinity glutamate uptake site ([3H]aspartate) using in situ radioligand binding with autoradiography and levels of mRNA for kainate receptors using in situ hybridization in the dorsolateral prefrontal cortex from 20 subjects with schizophrenia and 20 controls matched for age and sex. Levels of [3H]kainate binding were significantly decreased in cortical laminae I-II (p = 0.01), III-IV (p < 0.05), and V-VI (p < 0.01) from subjects with schizophrenia. By contrast, levels of [3H]MK-801, [3H]AMPA, [3H]aspartate, or [3H]CGP39653 binding did not differ between the diagnostic cohorts. Levels of mRNA for the GluR5 subunit were decreased overall (p < 0.05), with no changes in levels of mRNA for GluR6, GluR7, KA1, or KA2 in tissue from subjects with schizophrenia. These data indicate that the decreased number of kainate receptors in the dorsolateral prefrontal cortex in schizophrenia may result, in part, from reduced expression of the GluR5 receptor subunits. PMID- 15886720 TI - Olanzapine attenuates the okadaic acid-induced spatial memory impairment and hippocampal cell death in rats. AB - It is hypothesized that olanzapine, an atypical antipsychotic drug, has beneficial effects on cognitive impairment and neuropathological changes in treating neurodegenerative diseases. In the present study, we investigated the effects of chronic administration of olanzapine on the spatial memory impairment and hippocampal cell death induced by the direct injection of okadaic acid (OA), a potent neurotoxin, into the rat hippocampus. After being pretreated with olanzapine (0.5 or 2 mg/kg/day, i.p.) for 2 weeks, the rats were unilaterally microinjected with OA (100 ng) into the hippocampus, and then were continuously administrated with olanzapine for an additional week The rats were trained on a spatial memory task in an eight-arm radial maze before OA administration, and tested on the same task 18 h after the last olanzapine injection. After the behavioral test, the rats were killed for Nissl staining and terminal deoxynucleutidyl transferase-mediated biotinylated UTP nick end labeling staining. OA significantly induced spatial memory impairment, and caused pyramidal cell loss in the CAI and apoptotic cell death in the hippocampus. Olanzapine significantly attenuated OA-induced spatial memory impairment and the OA-induced neuropathological changes in the hippocampus. These findings suggest that olanzapine may have therapeutic effects in treatment of cognitive impairment and neuropathological changes of schizophrenia and other neurodegenerative diseases. PMID- 15886721 TI - Cortisol and cytokines in chronic and treatment-resistant patients with schizophrenia: association with psychopathology and response to antipsychotics. AB - The bilateral communication between the immune and neuroendocrine systems plays an essential role in modulating the adequate response of the hypothalamic pituitary-adrenal (HPA) axis to the stimulatory influence of cytokines and stress related mediators. Growing evidence suggests that neuro-immune-endocrine crosstalk may be impaired in schizophrenia. We determined the relationship between cortisol, cytokines interleukin-2 (IL-2) and interleukin-6 (IL-6), and symptoms in schizophrenia during treatment with typical and atypical antipsychotic drugs. Subjects included 30 healthy controls (HC) and 78 schizophrenic (SCH) in-patients. SCH were randomly assigned to 12-week treatment with 6 mg/day of risperidone or 20 mg/day of haloperidol using a double-blind design. Clinical efficacy was determined using the Positive and Negative Syndrome Scale (PANSS). Serum cortisol and IL-2 levels were assayed by radioimmunometric assay, and serum IL-6 levels by quantitative enzyme-linked immunosorbent assay. Following a 2-week washout period, serum levels of cortisol, IL-2, and IL-6 were increased in patients with schizophrenia compared to HC. Elevations in cortisol were associated with increase in both IL-2 and IL-6 in SCH. Moreover, elevations in cortisol were associated with negative symptoms and IL-2 with positive symptoms. In all, 12 weeks of risperidone treatment significantly decreased elevated cortisol and improved negative symptoms, but produced similar effects on IL-2 and IL-6 as well as on positive symptoms compared to haloperidol. The improvement of negative symptoms was related to the change in cortisol. Our results suggest that the imbalance in the HPA axis and cytokine system in patients with SCH is implicated in clinical symptoms, and is improved with atypical antipsychotic treatment. PMID- 15886722 TI - Platelet serotonergic markers as endophenotypes for obsessive-compulsive disorder. AB - Although compelling evidence has shown that obsessive-compulsive disorder (OCD) has a strong genetic component, its genetic basis remains to be elucidated. Identifying biological abnormalities in nonaffected relatives is one of the strategies advocated to isolate genetic vulnerability factors in complex disorders. Since peripheral serotonergic disturbances are frequently observed in OCD patients, the aim of this study was to investigate if they could represent endophenotypes, by searching for similar abnormalities in the unaffected parents of OCD patients. We assessed whole blood serotonin (5-HT) concentration, platelet 5-HT transporter (5-HTT) and 5-HT2A receptor-binding characteristics, and platelet inositol trisphosphate (IP3) content in a sample of OCD probands (n = 48) and their unaffected parents (n = 65), and compared them with sex- and age matched controls (n = 113). Lower whole blood 5-HT concentration, fewer platelet 5-HTT-binding sites, and higher platelet IP3 content were found in OCD probands and their unaffected parents compared to controls. Whole blood 5-HT concentration showed a strong correlation within families (p < 0.001). The only parameter that appeared to discriminate affected and unaffected subjects was 5-HT2A receptor binding characteristics, with increased receptor number and affinity in parents and no change in OCD probands. The presence of peripheral serotonergic abnormalities in OCD patients and their unaffected parents supports a familial origin of these disturbances. These alterations may serve as endophenotypic markers in OCD, and could contribute to the study of the biological mechanisms and genetic underpinnings of the disorder. PMID- 15886723 TI - A comparison of brain and serum pharmacokinetics of R-fluoxetine and racemic fluoxetine: A 19-F MRS study. AB - Racemic fluoxetine consists of R- and S-fluoxetine, which are metabolized to R- and S-norfluoxetine, respectively. This study was designed to compare brain levels achieved with R-fluoxetine to those achieved with racemic fluoxetine in healthy subjects using fluorine-19 (19-F) magnetic resonance spectroscopy (MRS). In all, 13 healthy volunteers received study drug for 5 weeks using a dosing schedule designed to achieve steady state for 20 mg/day racemic fluoxetine, 80 mg/day R-fluoxetine, or 120 mg/day R-fluoxetine. The resulting brain drug levels were measured using 19-F MRS. At 5 weeks, the racemate, 80 and 120 mg/day R fluoxetine groups had mean brain levels of 25.5, 34.9, and 41.4 microM, respectively. In the serum, R-norfluoxetine, which is thought to be an inactive metabolite, accounted for 17, 71, and 63% of the fluoxetine/norfluoxetine concentration, respectively. When the relative proportion of active to total species in serum are taken into account, the data suggest that doses of R fluoxetine greater than 120 mg/day would be needed to achieve brain levels of active drug comparable to 20 mg/day of racemate. The 120 mg/day R-fluoxetine group experienced a mean increase in QTc interval of 44 ms, with one individual having an increase of 89 ms, which suggests that higher doses may not be tolerable. While these data support the use of MRS to aid in defining the therapeutic dose range for drug development, they also highlight the need for additional studies with concurrent animal models to establish the validity of using serum drug/metabolite ratios to interpret MRS determined brain drug levels. PMID- 15886724 TI - Restraint increases dopaminergic burst firing in awake rats. AB - In anesthetized animals, dopamine neurons fire in tonic and phasic firing modes hypothesized to be regulated by dissociable circuit mechanisms. Salient events critical to learning, reward processing, and attentional selection elicit transient phasic bursts. It is unclear, however, how burst activity contributes to sustained firing patterns in awake animals and if behavioral conditions known to affect dopaminergic neurotransmission change impulse activity levels. Acute stress is known to increase extracellular dopamine in the striatum and the prefrontal cortex. In this study, we have used multiunit recording to define and follow activity patterns in single dopaminergic neurons across days and to determine how restraint, a model of acute stress, changes tonic and phasic firing patterns. Long-term recording shows that a population of 23 putative dopamine neurons has heterogeneous firing profiles under baseline conditions. In all, 62% showed significant burst activity under resting conditions, while others showed predominantly regular (17%) or random (21%) activity patterns. Restraint increased mean firing rate in all dopamine neurons, but preferentially increased burst firing in neurons with higher burst rates under resting conditions. Finally, we show that increased burst firing can persist 24 h after a single exposure to stress. These data indicate that subsets of dopamine neurons may be sensitive to circuit mechanisms activated by stress and that persistent changes in burst firing may be evidence of synaptic plasticity. Furthermore, increased burst firing may be a mechanism through which stress augments extracellular dopamine in selected terminal regions. PMID- 15886725 TI - Knowledge and attitudes about influenza vaccination amongst general practitioners, practice nurses, and people aged 65 and over. AB - AIMS: To identify knowledge, attitudes, and beliefs influencing influenza immunisation coverage in people aged 65 years and over in New Zealand METHODS: A postal survey of general practitioners (GPs) and practice nurses (PNs) was carried out during 2001-2002 in four regions of New Zealand (Northland, Waikato, Bay of Plenty, and Christchurch) with low or high influenza-immunisation coverage, based on 2000 data. A telephone survey of people aged 65 and over was also carried out in each region. Both surveys assessed knowledge and attitudes about influenza and influenza vaccination, including barriers to vaccination and personal vaccination status. RESULTS: GPs, PNs, and people aged 65 and over were generally well-informed about influenza, its complications, and the effectiveness of influenza immunisation. Some misinformation, however, is still prevalent in people 65 and over, and these beliefs discourage some older people from being immunised. CONCLUSIONS: Influenza vaccination coverage among high-risk groups in New Zealand is suboptimal. Overseas studies have shown that patient attitudes and beliefs influence influenza vaccination uptake, and our findings support this. While levels of awareness about influenza and the role of vaccination in its control are high, this study suggests a need for information specifically targeted towards younger age groups about the benefits of vaccination in healthy older adults. PMID- 15886726 TI - Twelve-month experience of acute stroke thrombolysis in Christchurch, New Zealand: emergency department screening and acute stroke service treatment. AB - AIMS: To determine the safety and efficiency of an acute stroke thrombolysis service in a New Zealand public hospital setting. METHODS: A 12-month audit of patients referred to the Christchurch Hospital Stroke Thrombolysis Service (STS) between 1 April 2002 and 31 March 2003 was undertaken. RESULTS: Sixty-one patients were referred to the STS during the study period, of whom 16 were treated with tissue plasminogen activator (t-PA). For treated patients, the median time from stroke onset to hospital presentation was 60 minutes, 'door-to CT' time was 60 minutes, and the 'door-to-needle' time was 99 minutes. Minor protocol violations were recorded in two patients, but did not influence outcome. No patient was treated after 3 hours of stroke onset. Intracerebral haemorrhage occurred in two patients: one patient was significantly improved compared with pre-treatment status; a minor temporary deterioration occurred in the other patient. Eight of 16 patients had improved by 4 or more points on the NIH Stroke Scale Score at 24 hours. CONCLUSIONS: Acute stroke thrombolysis can be delivered safely and in accordance with internationally accepted guidelines using the Christchurch Hospital STS model of emergency department screening and acute stroke service treatment. Further improvements in performance of the STS remain possible. PMID- 15886727 TI - Cassandra calling? PMID- 15886728 TI - Tissue plasminogen activator approved for stroke. PMID- 15886729 TI - Is PHARMAC's sole-supply tendering policy harming the health of New Zealanders? PMID- 15886730 TI - Guideline recommendations for the management of patients admitted with acute stroke: implications of a local audit. AB - AIM: To assess the feasibility of implementing the New Zealand guideline for management of stroke by auditing the gap between recommended care and that provided in Hawke's Bay Hospital (HBH). METHODS: Fifty randomly selected records of patients discharged with acute stroke between 1 June and 30 November 2003 were retrospectively reviewed using an audit tool developed from the guideline. RESULTS: Eight patients (16%) with non stroke syndromes were incorrectly diagnosed or coded as stroke on the basis of computed tomography (CT) scan reports. Brain imaging compliance was 100%; 86% within first 48 hours. Aspirin use or documented contraindications occurred in 62% patients within 48 hours, although the delay after imaging averaged 12.55 hours. At discharge, aspirin compliance was 100% in the surviving 42 patients. Twenty-nine (69%) patients were discharged on antihypertensive therapy, but two patients were readmitted with hypotension. Compliance with recommended multidisciplinary assessments within 48 hours included swallowing (88%), mobility (88%), communication (78%), and self care ability (60%). Patients not admitted directly to the stroke unit (19, 38%) were less likely to receive recommended interventions. Compliance with recommended predischarge assessments included suitability of accommodation (100%) and home supports (92%), but only 19% received documented advice about driving. CONCLUSIONS: Management of inpatients with acute stroke in HBH is close to most of the guideline recommendations. The identified deficiencies in patient care are potentially easily rectifiable and full implementation of the guidelines is feasible. PMID- 15886731 TI - New Zealand's psychiatry of old age services. Revisiting 'the view from the bottom of the cliff'--have we made any progress since 1998? AB - AIM: In 1998, the New Zealand Branch of the Faculty of Psychiatry of Old Age (FPOA) surveyed psychiatry of old age (POA) services in New Zealand for their resource levels. At that time, they were low by international and Mental Health Commission 'Blueprint' levels. In 2003, POA services in New Zealand were resurveyed to determine how much progress had been made in the ensuing 5 years. METHOD: All POA services in New Zealand who had Faculty members were surveyed for information on their service's administrative structure; access to acute, long stay and day hospital places; staffing levels of the different disciplines; the range of services they provided, and what they considered were strengths, gaps and potential improvements. RESULTS: Thirteen services responded, providing services for 387,000 elderly people. Since 1998, acute bed numbers and day hospital places had proportionately decreased nationwide, with disparity of access across the country. Inpatient nursing staff had decreased by 34%, with community nurse numbers barely increasing. There were modest increases in the number of psychiatrists, junior doctors, clinical psychologists, and social workers. CONCLUSIONS: There has been little progress towards achieving 'Mental Health Commission Blueprint' benchmark levels of service of 1998, and nationally there has been a decrease in access to acute beds, without a concomitant increase in staff to support older people in community settings. PMID- 15886732 TI - Quality of residential care for older people: does education for healthcare assistants make a difference? AB - AIM: To determine the impact of a healthcare assistant education programme on the quality of care for older people living in a residential home in Auckland, New Zealand. METHOD: A pre- and post-intervention evaluation study was undertaken within a residential home for older people. Quality of care was established by two periods of non-participant time-sampling observation of residents, separated by a programme of 10 interactive teaching sessions for healthcare assistants. Informed consent was gained from all observed residents and staff. RESULTS: Using the Quality Assessment Project (QAP) quality measure, the non-participant time sampling observation data identified a 12.5% increase in resident care that was considered appropriate and adequate (chi-squared=12.05) and an 11.53% decrease in resident care that was considered inappropriate and inadequate (chi squared=11.43). The intermediate grades did not alter significantly. Residents with low functional activity scores (Barthel Index) received better care after the education intervention (chi-squared=32.99), as did residents with moderate cognitive impairment (Abbreviated Mental Test Score [AMTS]<8/10). CONCLUSION: Healthcare assistant education can positively impact on the quality of care given to older people in residential care. PMID- 15886733 TI - Shared care between geriatricians and orthopaedic surgeons as a model of care for older patients with hip fractures. AB - AIMS: To evaluate the effect of shared care between geriatricians and orthopaedic surgeons as a model of care for older patients with hip fractures. METHODS: All patients over the age of 65 years are under the shared care of an orthopaedic surgeon and geriatrician (the Ortho-Medicine Service) when they are admitted to the Orthopaedic Service, Christchurch Hospital, New Zealand. This retrospective case records audit includes all patients over the age of 65 years with hip fracture admitted to this service over a 6-month period from December 2002 to June 2003. RESULTS: There were 150 patients. The median age was 83 years (range 66-99 years). Median total length of stay was 23 days. Median time delay until theatre was 43.5 hours. Inpatient mortality was 0.7%. Of 97 patients admitted from home, 86(88.6%) returned home, 6 (6.2%) went to rest home care, and 5 (5.2 %) went to hospital level care. Of 43 patients admitted from rest home care, 40 (93%) returned to rest home care, and 3 (7.0 %) were discharged to hospital level care. Three patients admitted from rest home dementia care and six patients admitted from hospital level care were discharged back to their pre-morbid place of domicile. At discharge, 86.8% of patients were on Vitamin D supplementation and over 80% were on calcium. Only 10.6% were discharged on bisphosphonates. CONCLUSIONS: Shared care between geriatricians and orthopaedic surgeons for older people with hip fractures is associated with a low in-patient mortality, with the majority returning to their pre-morbid place of domicile. Length of stay has increased. Most patients are discharged on treatment for osteoporosis. PMID- 15886734 TI - Dinner Bed and Breakfast for Older People: early experiences of a short-term service to manage acute hospital demand. AB - AIMS: The paper describes a service (Dinner Bed and Breakfast [DBB]) to provide short-term rest home care to acutely unwell elderly people as an alternative to acute hospital admission. This was part of a larger project to manage acute general hospital demand. Service changes were introduced after an initial audit cycle and key outcomes monitored. METHOD: Retrospective audit of patient data for three audit periods. RESULTS: The interim outcomes (at conclusion of DBB funding) in the initial audit were 46% of patients able to return home, 32% remained in the rest home, and 22% were admitted to hospital. The proportion of patients returning home from DBB increased to 68% over the study period. At final outcome (at end of index illness), the return home rate increased from 73% to 85% and the number requiring permanent rest home care decreased from 22% to 14%. CONCLUSIONS: Short-term rest home care may be a viable alternative to acute hospital care, but the service needs to include appropriate patient selection, multidisciplinary care, and ongoing monitoring of patient outcomes. One of the risks of this service is patients staying on in rest home care. Short-term enhanced home care may be preferable to rest home care to avoid this risk. The shift of care (from acute hospital to community) caused strain on specialist geriatric services, which was not planned or funded. PMID- 15886735 TI - An evaluation of two respite models for older people and their informal caregivers. AB - AIM: To evaluate two case-management models of respite relief care at Waitemata District Health Board. METHOD: The evaluation consisted of semi-structured interviews and postal surveys for clients utilising respite care and staff members involved in both the North/West and Rodney models of respite care in Auckland, New Zealand. Across the two regions, a total of 2 older people and their informal caregivers, 2 respite coordinators, and the Needs Assessment Service Coordination (NASC) Manager were interviewed. In addition, postal surveys were received from 21 older people, 36 informal caregivers, 11 NASC workers, and 3 allied health professionals. RESULTS: Findings revealed there was generally high satisfaction with both the respite models. Caregivers believed the respite service did give them a break, although it was insufficient. Caregivers reported concerns regarding how respite facilities could improve and the older person's deterioration post respite. Staff identified improvements for each of the models. CONCLUSIONS: The respite models are flexible and provide control in decision making for informal caregivers leading to an increase in choice for home-based respite. Case-managers need to form strong partnerships with caregivers to ensure that the needs of both the informal caregiver and older person are met. Short term placement in residential respite facilities for respite care may be linked to worsening functional ability of the older person. PMID- 15886736 TI - Salamol asthma inhaler fails roadside alcohol breath testing. PMID- 15886737 TI - Colo-colonic intussusception secondary to a colonic lipoma diagnosed with preoperative CT scan. PMID- 15886738 TI - Valuing prevention: discounting health benefits and costs in New Zealand. PMID- 15886739 TI - Faecal calprotectin: the case for a novel non-invasive way of assessing intestinal inflammation. PMID- 15886740 TI - Residential care workers and residents: the New Zealand story. AB - AIMS: To describe the nature and size of long-term residential care homes in New Zealand; funding of facilities; and the ethnic and gender composition of residents and residential care workers nationwide. METHODS: A postal, fax, and email survey of all long-term residential care homes in New Zealand. RESULTS: Completed surveys were received from an eligible 845 facilities (response rate: 55%). The majority of these (54%) facilities housed less than 30 residents. Of the 438 (94%) facilities completing the questions about residents' ethnicity, 432 (99%) housed residents from New Zealand European (Pakeha) descent, 156 (33%) housed at least 1 Maori resident, 71 (15%) at least 1 Pacific (Islands) resident, and 61 (13%) housed at least 1 Asian resident. Facilities employed a range of ethnically diverse staff, with 66% reporting Maori staff. Less than half of all facilities employed Pacific staff (43%) and Asian staff (33%). Registered nursing staff were mainly between 46 and 60 years (47%), and healthcare assistant staff were mostly between 25 and 45 years old (52%). Wide regional variation in the ethnic make up of staff was reported. About half of all staff were reported to have moved within the previous 2 years. CONCLUSIONS: The age and turnover of the residential care workforce suggests the industry continues to be under threat from staffing shortages. While few ethnic minority residents live in long-term care facilities, staff come from diverse backgrounds, especially in certain regions. PMID- 15886741 TI - Raloxifene for hormone replacement therapy (HRT). PMID- 15886742 TI - Single embryo transfer in IVF treatment. PMID- 15886743 TI - Is the Internet as essential as a stethoscope for patient care? PMID- 15886744 TI - [Alzheimer's dementia, sleep disorders and nuclear medicine]. AB - In case new diagnostic procedures for Alzheimer's dementia (AD) appear, Nuclear Medicine (NM) would like to be aware of them in order to evaluate its own contribution to diagnose AD by SPET and PET brain studies. Recently, sleep disturbances were studied in AD and tend to be diagnostic for early AD. In AD the actual time of night sleep was found to be 5.7 h, while awakeness time for the same night sleep increased to 2.7 h. Also in AD, the REM and the slow wave stage (SWS) during sleep are shorter and hypopnea and apnea phases are abundant. Internal body temperature during night sleep is only slightly increased in AD, while in temporofrontal dementia and in normal individuals this increase is significant. The circadian rhythm of melatonin is disturbed in AD. The normal duration of inspiration and expiration during daytime which is reversed during normal night sleep, has not been studied in patients with AD. However, this reverse condition favoring inspiration is expected to provide more oxygen to the brain. Chronic but not acute stress causes memory loss and is currently being studied by us as a possible causative factor for memory loss in AD. Tomographic SPET and PET brain studies can locate the site of brain damage in AD. This is important since memory has recently been classified into four categories, namely episodic, semantic, procedural and working memory. In early AD only procedural memory remains intact. This means that these patients may drive a car, do computer word processing and play some games at home or/and in the field. This memory is located in specific nuclei in the cerebellum and the occipital frontal area which do not relate to sites of other kinds of memory. This difference could be well identified by tomographic SPET or PET studies. Thus NM may also diagnose the early stage of AD. Another issue refers to the indications that the unified Medicare and Medicaid system in the USA has issued on September 15, 2004 for performing a PET (18)F-FDG study for AD. These indications are fully described in this editorial. PMID- 15886745 TI - [Angiogenesis in cancer and its detection with radiolabeled biomolecules]. AB - The present article refers to the mechanism of angiogenesis, a phenomenon which in certain cases is normal, however it usually accompanies the formation of solid cancerous tumors. During the process of angiogenesis, migration, differentiation and proliferation of endothelial cells occurs. In some pathological cases, the most important of which is cancer, the mechanism of angiogenesis is reinforced. It has been observed that a tumor cannot grow without the formation of new blood vessels in the surrounding tissues, which account for blood supply to the tumor. The process of angiogenesis is regulated by chemical signals of the organism, which function as an "angiogenesis switch" regulating the formation of new vasculature. A variety of anti-angiogenic agents which lead to angiogenesis inhibition are found in the clinical trial phase. Among these agents are: i) molecules which inhibit the action of Vascular Endothelial Growth Factors, VEGF, ii) molecules which obstruct migration, differentiation and proliferation of endothelial cells, via their binding to receptors of the alpha(nu)beta(3) integrins and iii) inhibitors of metalloproteinases (MMP). Certain molecules of the above mentioned categories, labeled with radionuclides which emit gamma- radiation or beta- particles or positrons, have been proposed and are being evaluated as possible radiopharmaceuticals, for the visualization of receptors involved in the mechanism of angiogenesis, aiming at the scintigraphic detection of primary or metastatic cancer at an early stage and possibly, aiming at its inhibition/treatment. Regarding the study of angiogenesis the following have been described: a. antibodies targeting VEGF, labeled with radionuclides emitting beta and/or gamma-radiation, which can be applied for the diagnosis and possibly, for the treatment of cancer, b. peptide derivatives which contain the amino-acid sequence RGD (Arg-Gly-Asp) and compete for the alpha(nu)beta(3) integrins, with the proteins of the stroma. It has been found that these radiolabeled derivatives localize in tumors and can be used for the visualization and, possibly, for the tumor eradication of primary and metastatic cancer. PMID- 15886746 TI - [Current trends in infection imaging]. AB - Infection imaging is a field of intense interest in medicine because, despite the great advances in our understanding of microorganisms and the development of new antimicrobial agents, infection remains a major cause of patient morbidity and mortality. Nuclear Medicine for many years has an important role in this field. The significance of this role varies in different clinical situations, in some cases being complementary to other imaging modalities as in postoperative patients, and in some cases being the method of choice as in orthopaedic infections after implants have been placed. Today there are many agents in Nuclear Medicine capable in imaging infection. Each of them has its own advantages and drawbacks. In recent years there is an increased research in the field with new methods tested. Among them are labelled antibacterial agents, labelled antimicrobial peptides, monoclonal antibodies for leukocyte labelling, labelled liposomes, and (18)F-FDG-PET. In this short article there is an effort to briefly review these agents and their possible role in infection imaging. PMID- 15886747 TI - [Evaluation of a quantitative diagnostic sacroiliac bone scan index in cases of chronic low back pain in young male adults]. AB - Quantitative bone scan of the sacroiliac joints has long been an established diagnostic method in cases of chronic low back pain (LBP), though its value has been questioned due to the significant overlap of the numerical values between patients with inflammatory sacroiliitis and healthy controls. In an effort to solve this dissent, 133 young male adults aged 18-36 years were studied. We thus aimed to have a relatively homogenous population sample that would include the age that many sacroiliac diseases appear. Thirty-two of our patients had chronic, inflammatory disease of non-infectious origin, as tested by clinicolaboratory procedures (Group A), 29 had mechanical (non-inflammatory type) LBP (Group B), and 72 had been scintiscanned for reasons irrelevant to spine disease (Group C). The members of each group were also classified in three subgroups (a, b and c) according to age. The protocol of planar static multispot bone scan was applied to all three Groups. Three regions of interest (ROI), two on the sacroiliac joints plus one over the L4 vertebra were drawn and finally a non-dimensional numerical parameter called "lambda" was extracted by the equation lambda=total counts/total pixels of the "hottest" of the two sacroiliac joints area divided by the counts/pixels corresponding to the L4 values (lambda=SI/L4). The statistical analysis showed negative correlation of lambda with age in all three groups (P~0.04 for Group A, P~0.012 for Group B and P~0.05 for Group C). When all Groups were examined regardless of age, lambda appeared significantly different (P<0.0005) between Groups A and C, as well as between Groups A and B (P~0.002) but there was no difference between Groups B and C (P~0.12). When the members of each group were analyzed according to age, the paired difference of lambda stirred with remarkable vagueness along the whole spectrum of statistical significance. Conclusively, lambda seems to decrease with ageing at ages ranging from 18 to 36 years (P/=50% stenosis or occlusion of at least one coronary artery, who were scheduled to undergo PTCA. A gamma-camera gated single photon emission tomography myocardial perfusion scintiscan was performed as a baseline study with (99m)Tc-sestamibi (GSPET-B) and another similar scintiscan after nitrate augmentation (GSPET-N) before PTCA and two to six months after PTCA (GSPET-R), to assess the extent of perfusion defects, contraction abnormalities and myocardial viability. Cedars QGS software was used for semi quantitative assessment and sum perfusion scores were calculated for each study. According to our results from the 174 hypoperfused segments studied by GSPET-B, 137 segments had tracer activity <50%. From the 137 segments with tracer activity of <50% only 51 (37%) remained unaltered after PTCA. Twenty-six of them (51%) were described as nonviable after the GSPET-N study and the remaining 25 were defined as viable. Our study demonstrated significant perfusion improvement after nitrate augmentation (mean sum perfusion score decreased from 34+/-9 in the baseline study (SBS) to 23+/-11 in the GSPECT N study (SNS), P=0.04. There was no significant difference between SNS and mean sum perfusion score after revascularisation (SRS), 23+/-11 and 24+/-13 respectively (P=0.833). The specificity, sensitivity and accuracy of the perfusion improvement after PTCA, were calculated as: 52%, 85% and 76% respectively. The low specificity in our study could be due to performing GSPET-R in some patients, six months after PTCA; during this time restenoses may occur. Two of our patients whose perfusion and myocardial wall motion kinetics had worsened and also had clinical symptoms of pain and fatigue, were considered to have developed restenosis. In the present study, myocardial wall motion kinetics showed non-significant improvement of global ejection fraction (EF). EF increased from 43.9%+/-3.3% to 48.9% after nitrate augmentation (P=0.262) and to 47.2%+/ 6.4% after revascularization (P=0.091). Myocardial wall motion hypokinesia showed significant improvement of severity scores in the GSPET-N study, as well as after PTCA revascularization (P<0.01). It is concluded that GSPET-N (99m)Tc-sestamibi imaging significantly improves the detection of defect reversibility. On the basis of our results, it appears appropriate to recommend GSPET-N (99m)Tc sestamibi imaging only in patients with perfusion defects and tracer activity of <50%. In such cases it is recommended to perform GSPET not only for perfusion but also for a myocardial wall motion kinetic study. The follow up study to evaluate the result of PTCA is recommended to be performed within 2 months after PTCA, before restenosis may occur and 6 months after PTCA if restenosis is suspected. PMID- 15886755 TI - Cytokinesis-block micronucleus test in patients undergoing radioiodine therapy for differentiated thyroid carcinoma. AB - Cytokinesis-blocked micronucleus assay (MNT) was applied in the peripheral blood lymphocytes of patients undergoing radioiodine-131 ((131)I) therapy for differentiated thyroid carcinoma (DTC) after thyroidectomy to assess the genotoxic risk of this therapy. The amount of administered (131)I activity varied from 3,330 to 4,030 MBq. Blood samples were taken immediately before (131)I administration and one month later, along with routine blood tests. Twenty-six patients underwent radioiodine ablation (RIA) or radioiodine therapy (RIT) after radical thyroidectomy. The aim of this therapy was to ablate residual thyroid tissue, to treat lymph node metastases and/or distant lung metastases. The amount of orally administered activity of (131)I ranged between 3,330 and 4,030 MBq according to the kind of therapy (RIA or RIT). In five patients the cytogenetic analysis was performed immediately before and one month after the second therapy which was given to them 6 months to 1 year after the first. Three patients were male and 23 female. The age of the patients ranged between 23 and 76 years (mean age: 48.6 years). Results show that after radioiodine therapy there is a significant increase in the frequency of micronuclei. Comparing the average frequency of micronuclei in the patients studied before and after (131)I, a more than doubling increase was found. Mean values +/- SD of the patients before and after (131)I therapy were 10.72 per thousand +/- 5.84 per thousandand 25.28 per thousand +/- 12.6 per thousand respectively. These findings indicate a genotoxic activity of (131)I therapy estimated after a period of one month. PMID- 15886756 TI - Cutaneous radiation necrosis as a complication of yttrium-90 synovectomy. AB - Radiation synovectomy is an effective method of treatment of chronic synovitis in patients with conditions such as rheumatoid arthritis, haemophilic synovitis and pigmented villonodular synovitis. This case demonstrates one of the potential complications of this procedure in a 46 year old man treated with bilateral radiation synovectomy for haemophilic synovitis. Cutaneous radiation necrosis is a known but rare complication of this procedure and this case is reported to demonstrate this known complication and to highlight that appropriate technique is required to avoid this. PMID- 15886757 TI - Subspecialities in nuclear medicine; a requirement. PMID- 15886758 TI - The future of nuclear medicine. PMID- 15886759 TI - Prospectives in nuclear medicine. PMID- 15886760 TI - Combination of orthogonal supramolecular interactions in polymeric architectures. AB - Supramolecular polymers represent a highly interesting approach towards new "smart materials". A recent strategy includes the combination of different "orthogonal" non-covalent binding sites within one polymer system. Different functionalities can be introduced in a highly defined way by controlled self assembly processes. This feature article presents highlights in the supramolecular polymer chemistry of multiple hydrogen-bonding, metal complexation (especially of bi- and terpyridines) and host-guest interactions as well as recent advances in combining these interactions in novel polymers. PMID- 15886761 TI - Synthesis and crystal structure of a push-pull quinoidal porphyrin: a nanoporous framework assembled from cyclic trimer aggregates. AB - A quinoidal porphyrin has been synthesised with such a curved pi-system that pi pi stacking leads to the formation of cyclic trimer aggregates in the crystal, which pack to generate cylindrical channels with an internal diameter of 1.0 nm. PMID- 15886762 TI - Porous lanthanide-organic framework with zeolite-like topology. AB - Two novel isomorphous porous frameworks of Ln(III), {[Ln2(imidc)2(H2O)3](H2O)}n, [Ln = Gd(III) (1) and Er(III) (2)] have been synthesized hydrothermally using multifunctional 4,5-imidazoledicarboxylic acid (imidc) as a connector; they have a zeolite-like network topology which can provide another form upon dehydration, and exhibit selective adsorption properties for H2O over N2, CO2 and MeOH. PMID- 15886763 TI - Sorting of polymorphs based on mechanical properties. Trimorphs of 6-chloro-2,4 dinitroaniline. AB - Mechanical shearing of crystals of a polymorph with a layered structure may be used to separate them from harder crystals of a visually indistinguishable polymorph that resist such shearing. PMID- 15886764 TI - Head-to-tail self-assembly of a calix[4]arene inclusion polymer controlled by a pendant arm. AB - A calix[4]arene functionalized at one phenolic group with a pendant ethoxy acetate group, forms an inclusion complex that is stable even in the presence of other potential guest molecules. PMID- 15886765 TI - A novel self-assembled organic tubular structure. AB - A novel organic tubular structure with the walls consisting of aromatic rings was constructed by the self-assembly of a two-dimensional sheet-like molecule in the solid state. PMID- 15886766 TI - Microfabrication of encoded microparticle array for multiplexed DNA hybridization detection. AB - A strategy for the high-sensitivity, high-selectivity, and multiplexed detection of oligonucleotide hybridizations has been developed with an encoded Ni microparticle random array that was manufactured by a "top-down" approach using micromachining and microfabrication techniques. PMID- 15886767 TI - The step-wise assembly of an undecanuclear heterotrimetallic cyanide cluster. AB - The step-wise assembly of the high nuclearity cluster, {[Ni(II)(H2O)5]6[Co(III)(tmphen)2]3[Fe(II)(CN)6]2}13+, is achieved by treating {[Co(tmphen)2]3[Fe(CN)6]2} with six equivalents of Ni(ClO4)2 in aqueous MeOH. PMID- 15886768 TI - Nanotubes with the TiO2-B structure. AB - Here we report the first synthesis of TiO2-B nanotubes by a simple hydrothermal route; lithium may be intercalated up to a composition of Li(0.98)TiO2 compared with Li(0.91)TiO2 for the corresponding nanowires. PMID- 15886769 TI - Thermal solid state synthesis of coordination complexes from hydrogen bonded precursors. AB - Thermal dehydrochlorination of crystalline 4-picolinium salts of [PtCl4]2- and [PdCl4]2- leads to formation of trans-[MCl2(4-picoline)2](M = Pt, Pd). PMID- 15886770 TI - Nickel-catalyzed cocyclotrimerization of arynes with diynes; a novel method for synthesis of naphthalene derivatives. AB - The NiBr2(dppe)-Zn system effectively catalyzes the [2 + 2 + 2] cocyclotrimerization of arynes with diynes, leading to substituted naphthalene derivatives in moderate to good yields. This cocyclotrimerization reaction shows excellent tolerance of functional groups and leads to products of 5- to 7 membered fused-ring sizes. PMID- 15886771 TI - Benzene C-H activation by platinum(II) complexes of bis(2 diphenylphosphinophenyl)amide. AB - The amido diphosphine complexes [PNP]PtMe and [PNP]PtOTf, where [PNP]- is bis(2 diphenylphosphinophenyl)amide, effectively activate the benzene C-H bond in the presence of an appropriate Lewis acid or base, leading to the formation of [PNP]PtPh quantitatively. PMID- 15886772 TI - A new synthesis of dienone lactones using a combination of hypervalent iodine(III) reagent and heteropoly acid. AB - The oxidation of non-phenolic alkanoic acid derivatives to oxygen heterocycles was investigated; a new oxidative route to dienone lactones has been developed using a combination of hypervalent iodine(III) reagent, phenyliodine(III) bis(trifluoroacetate)(PIFA), and heteropoly acid (HPA). PMID- 15886773 TI - Model systems for flavoenzyme activity: a tuneable intramolecularly hydrogen bonded flavin-diamidopyridine complex. AB - We report the electrochemically tuneable intramolecular hydrogen bonding interactions between a covalently linked flavin-diamidopyridine unit. PMID- 15886774 TI - Supramolecular chirality control by solvent changes. Solvodichroic effect on chiral porphyrin aggregation. AB - The mode of aggregation of amphiphilised porphyrin derivatives bearing a chiral functionality is strongly affected by solvent composition; this results in the tuning of the supramolecular chirality of the porphyrin aggregates. PMID- 15886775 TI - Prediction of the 13C NMR chemical shifts of organic species adsorbed on H-ZSM-5 zeolite by the ONIOM-GIAO method. AB - The ONIOM-GIAO method has been used to accurately predict 13C NMR chemical shifts for a series of organic species adsorbed on H-ZSM-5 zeolite. This is useful for the spectroscopic identification of complicated catalytic systems. PMID- 15886776 TI - A synthesis of multisubstituted vinylsilanes via ynolates: stereoselective formation of beta-silyl-beta-lactones followed by decarboxylation. AB - (Z)-Selective synthesis of multisubstituted vinylsilanes was achieved by stereoselective protonation or alkylation of beta-silyl-beta-lactone enolates, prepared by cycloadditions of acylsilanes with ynolates, followed by decarboxylation. PMID- 15886777 TI - Cyclic boronium and borenium cations derived from borabenzene-pyridine complexes. AB - A new class of boronium and borenium cations are prepared by reaction of pyridinium hydrochloride with pyridine stabilized borabenzene derivatives. PMID- 15886778 TI - Diethylene glycol ether-linked 3,4,5-trihydroxybenzamides as triply branched dendritic anchors to CdSe/ZnS core/shell type nanoparticles: potential hydrophilic fluorescent probes. AB - Functionalized N-2-mercaptoethyl-gallamides bearing three or five hydroxyl units that are tethered with diethylene glycol ether(s) allow for transferring hydrophobically pyridine-capped CdSe/ZnS core/shell nanoparticles from an organic to an aqueous layer with intact fluorescent profiles. PMID- 15886779 TI - Correlation between molecular dipole moment and centrosymmetry in some crystalline diphenyl ethers. AB - The presence of a large molecular dipole moment in diphenyl ethers leads unequivocally to a centrosymmetric crystal structure. PMID- 15886780 TI - Extended wet-spinning can modify spider silk properties. AB - Contrary to expectation, we demonstrate that spider dragline silk spun experimentally under water displays greater stiffness and higher resilience compared to silk spun "naturally" into air. We suggest that this consequence of extended wet-spinning is due to increased molecular orientation resulting from extension of the mobile phase. PMID- 15886781 TI - Alkenylation of allylic alcohols using alkenylboron dihalides: a formal transition-metal free Suzuki reaction. AB - Carbon-carbon bond formation via substitution of an allylic hydroxide with stereodefined alkenyl groups using alkenylboron dihalides in the absence of transition metals. PMID- 15886782 TI - Double carbonylation of zirconocene-alkyne complexes. AB - Zirconocene-alkyne complexes prepared from Cp2ZrBu2, phosphines and alkynes reacted with CO to give double carbonylation products, 4-hydroxycyclobuten-1-one derivatives after hydrolysis. PMID- 15886783 TI - Synthesis and characterization of Rh(III) corroles: unusual reactivity patterns observed during metalation reactions. AB - A new approach to the synthesis of Rh(III) corrole complexes is developed and an unusual activation of C-C and C-N bonds is disclosed. PMID- 15886784 TI - Zeolite filled polydimethylsiloxane (PDMS) as an improved membrane for solvent resistant nanofiltration (SRNF). AB - Due to the shape-selectivity of the pores and the induced polymer crosslinking, zeolite filled elastomers are excellent solvent-resistant nanofiltration membranes with enhanced fluxes and retentions compared to commercial membranes, allowing use in non-polar solvents and at high temperatures. PMID- 15886785 TI - Highly fluorescent photochromic diarylethene in the closed-ring form. AB - A highly fluorescent diarylethene in the closed-ring form was synthesized by the oxidation of 1,2-bis(2-methyl-1-benzothiophene-3-yl)perfluorocyclopentene (BTF6). PMID- 15886786 TI - Immobilized metal ion-containing ionic liquids: preparation, structure and catalytic performance in Kharasch addition reaction. AB - Immobilized metal ion-containing ionic liquid catalysts were prepared by the reaction between silyl-functionalized imidazolium ionic molecules and surface silanol groups of silica, followed by addition of MnCl2, FeCl2, CoCl2, NiCl2, CuCl2, or PdCl2; only the immobilized copper catalyst, which has a sandwiched CuCl4(2-) moiety, was very active for the Kharasch reaction between styrene and CCl4. PMID- 15886787 TI - DNA mismatch detection using a pyrene-excimer-forming probe. AB - A pyrene-excimer-forming probe allowed the easy and sensitive detection of a single base mismatch in target DNA. This was due to the faster strand exchange rate compared to a fully-matched target. PMID- 15886788 TI - Catalytic enantioselective arylation of aldehydes: boronic acids as a suitable source of transferable aryl groups. AB - The catalytic enantioselective arylation of several aldehydes using boronic acids as the source of transferable aryl groups is described; the reaction is found to proceed in excellent yields and high enantioselectivities (up to 97% ee) in the presence of a chiral amino alcohol. PMID- 15886789 TI - Enantiospecific inclusion of chiral 1,2-dichloroethane rotamers in the crystal lattice of chiral square-pyramidal Cu(II) complexes with perfectly polar alignment of guest and host molecules. AB - The inclusion compounds, [CuL(1)2(H2O)].(P)-C2H4Cl2 and [CuL(2)2(H2O)].(M) C2H4Cl2(HL1 = N-(2-hydroxy-5-nitrobenzyl)-(R)-alpha-methylbenzylamine and HL2 = N (2-hydroxy-5-nitrobenzyl)-(S)-alpha-methylbenzylamine), crystallise in the non centrosymmetric space group C2; intermolecular hydrogen bonding leads to a perfectly polar alignment of both host and guest molecules with enantioselectivity. PMID- 15886790 TI - Vitamin K intake and stability of oral anticoagulant treatment. PMID- 15886791 TI - The role of neovascularisation in the resolution of venous thrombus. AB - Deep vein thrombosis (DVT) can give rise to chronic debilitating complications, which are expensive to treat. Anticoagulation, the standard therapy for DVT, prevents propagation, but does not remove the existing thrombus, which undergoes slow natural resolution. Alternative forms of treatment that accelerate resolution may arise from a better understanding of the cellular and molecular pathways that regulate the natural resolution of thrombi. This review will outline our current understanding of the mechanisms of thrombus resolution and the role of neovascularisation in this process. Novel experimental treatments that may one day find clinical use are also discussed. The process of thrombus resolution resembles wound healing. The mainly monocytic inflammatory infiltrate, which develops, is associated with the appearance of vascular channels. These cells may drive resolution by encouraging angiogenesis, which contributes to restoration of the vein lumen. Significant numbers of bone marrow-derived progenitor cells have also been found in naturally resolving thrombi, but their precise phenotype and their role in thrombus recanalisation is unclear. Enhanced thrombus neovascularisation and rapid vein recanalisation have been achieved in experimental models with proangiogenic agents. Recent reports of the role of bone marrow-derived progenitor cells in the revascularisation of ischaemic tissues suggest that it may be possible to obtain the same effect by delivering pluripotent or lineage specific stem cells into thrombus. These cells could contribute to thrombus recanalisation by expressing a variety of proangiogenic cytokines or by lining the new vessels that appear within the thrombus. PMID- 15886792 TI - Thiazolidinediones inhibit apoptosis and heat shock protein 60 expression in human vascular endothelial cells. AB - This study evaluated direct effects of peroxisome proliferatoractivated receptor gamma(PPARgamma) agonists, including thiazolidinediones (TZDs), on vascular cell apoptosis and related protein expression to test the hypothesis that these effects are dependent on i) the respective agent's structure and ii) endothelial cells' vascular origin. Exposure (48 h) of human umbilical vein endothelial cells (HUVECs, n=6) to up to 10 microM troglitazone (TRO), rosiglitazone, pioglitazone, and to up to 50 microM RWJ241947=MCC-555 (RWJ) inhibited (p<0.05) apoptosis by 8 25%, whereas 15-deoxy-Delta(12-14)-prostaglandin J(2) (PGJ(2)) triggered (50 microM: + 400%, p<0.05) endothelial cell death versus control (=100%). Moreover, RWJ (50 microM) completely abrogated TNF-alpha(2000 U/ml) and stearic acid (200 microM) induced apoptosis in HUVECs . Similar results were obtained in human adult (saphenous) vein- and aortic endothelial cells, the latter showing no anti apoptotic response to TRO. In HUVECs, TZDs' anti-apoptotic effects inversely correlated (r=-0.95, p<0.01) with increased (p<0.05) expression of the apoptosis inhibitor bcl-2, whereas PGJ(2)-induced apoptosis was associated with upregulation of c-myc (+447%) and E2F-1 (+339%). Additionally, TZDs (by 25-39%) and PGJ(2) (-70%) reduced (p<0.05) expression of heat shock protein 60 (hsp60) showing no correlation with apoptosis (r=0.14, n.s.). Modulation of apoptosis by PPARgammaagonists differs in endothelial cells dependent on their vascular origin and the agonists' structure. Thiazolidinediones' ability to reduce both, endothelial apoptosis and hsp60 expression could well add to beneficial vascular effects attributed to these oral antidiabetic drugs. PMID- 15886793 TI - Effect of plasminogen activator inhibitor-1 deficiency on nutritionally-induced obesity in mice. AB - Plasminogen activator inhibitor-1 (PAI-1) is the main physiological inhibitor of tissue-type (t-PA) and urokinase-type (u-PA) plasminogen activator. Recent studies in murine models have yielded apparently conflicting data on a potential role of PAI-1 in adipose tissue development and obesity. To reinvestigate this issue, we have rederived PAI-1 deficient (PAI-1(-/-)) and wild-type (WT) mice and generated true littermates in a 81.25% C57Bl/6: 18.75% 129 SV genetic background. Male 5-week-old PAI-1(-/-) and WT mice were kept on a high fat diet (20.1 kJ/g) for 15 weeks. Body weight gain was comparable for both genotypes, and at the time of sacrifice total body weights (39+/-1.1 versus 41 +/- 1.2 g) as well as the weights of subcutaneous (SC, 1,520 +/- 110 versus 1,480 +/- 110 mg) adipose tissue were not significantly different. In contrast, the gonadal (GON, 1,900 +/- 43 versus 1,510 +/- 86 mg, p < 0.005) tissue mass was larger in PAI-1(-/-) mice. Plasma levels of insulin, leptin, glucose, triglycerides, total, HDL and LDL cholesterol were comparable for both genotypes. Immunohistochemical analysis of SC and GON adipose tissues did not reveal differences in adipocyte size or number between both genotypes, whereas blood vessel density was also comparable for GON fat but lower in SC fat of WT mice. Thus, this study in littermate mice on high fat diet did not reveal an effect of PAI-1 deficiency on body weight, and a differential effect on SC and GON adipose tissue. PMID- 15886794 TI - Impact of adjunctive tirofiban administration on myocardial perfusion and mortality in patients undergoing primary angioplasty for ST-segment elevation myocardial infarction. AB - Several studies have shown that suboptimal myocardial perfusion may be observed despite optimal epicardial recanalisation in patients undergoing primary angioplasty for ST-segment elevation myocardial infarction (STEMI), resulting in unfavourable outcome. The aim of the current study was to evaluate the benefits in myocardial perfusion and mortality from adjunctive tirofiban administration in patients undergoing primary angioplasty for (STEMI). Atotal of 1,969 patients with STEMI treated by primary angioplasty represent the population of the current study. All clinical, angiographic and follow-up data were prospectively collected. Tirofiban was administrated in 481 patients (24.4%) (all before angioplasty). Tirofiban was associated with less distal embolisation (11.7% vs 16.1%, p = 0.048), better postprocedural MBG 3 (50.9% vs 39.7%, adjusted p < 0.0001) and a significant reduction in 1-year mortality (3% vs 6.4%, adjusted p = 0.045). The benefits in mortality were confirmed in all subgroups identified according to the quartiles of the propensity score. This study shows that, when compared to control group, adjunctive tirofiban before primary angioplasty for STEMI is associated with better myocardial perfusion and a reduction in 1-year mortality. PMID- 15886795 TI - B-domain deleted factor VIII is aggregated and degraded through proteasomal and lysosomal pathways. AB - Factor VIII (FVIII) processing within mammalian cells is demonstrated to be much less efficient than proteins of similar size. The deletion of the B-domain from FVIII improves the level of production, due partly to the increase in mRNA synthesis. We aimed to characterise the cellular fate and the intracellular processing of the FVIII molecule devoid of B-domain. A B-domain deleted factor VIII (BDD-FVIII) possessing a furin consensus cleavage site in the connecting segment between the heavy and the light chain, was produced in CHO cell line. In such cells, FVIII was retained as two single chain products from which a majority was aggregated. The two species were located in Triton X-100 soluble (for 60-80%) and insoluble fractions (for 20-40%). The incubation of the expressing cells with tunicamycin (5 mug/ml) and the treatment of the intracellular species with a mixture of Neuraminidase and N-glycosidase-F revealed that both intracellular species were N-glycosylated. Furin over-expression neither diminished the intracellular FVIII contents nor improved its extracellular production. Intracellular FVIII was degraded through both lysosomal and proteasomal pathways as evidenced by inhibitor treatments (e.g. NH(4)Cl, leupeptin, clasto-Lactacystin beta-lactone and MG-132), pulse-chase analysis and confocal observations. This study demonstrates that a BDD-FVIII expressed in CHO cells is inefficiently processed consecutively to intracellular aggregation, proteasomal degradation, and routage to lysosomes. PMID- 15886796 TI - Phospholipid vesicles interfere with the binding of antibody fragments to the light chain of factor VIII. AB - Factor VIII binds to phospholipid membranes through the C2 domain (S2173-Y2332). Residues M2199, F2200, L2251, L2252, V2223, W2313 and V2314 at the tips of beta hairpins and loops are thought to contribute to phospholipid membrane binding. Similarly, residues in the C2 domain of the homologous protein factor V forma phospholipid binding site, but residues in the A3 and C1 domains are also thought to contribute to membrane binding. Phage display technology was previously used to isolate factor VIII light chain specific single-chain variable domain fragments (scFv) from patients with factor VIII inhibitors. Phospholipid vesicles inhibited the binding of factor VIII to scFvs WR1 and WR16 (epitope : E2181 M2199) with half saturation values of 23 and 47 muM respectively. The single point mutant F2200A factor VIII light chain bound to WR1 and WR16 with a much lower affinity than wild type protein suggesting that residue F2200 is also included in the epitopes of these scFvs. Binding of factor VIII to C2-specific scFvs WR13 and EL14 (epitope : K2207-M2321) was not inhibited by phospholipid vesicles. Consistent with this, F2200A factor VIII light chain bound to these scFvs with the same affinity as the wild type protein. However, phospholipid vesicles also inhibited the binding of factor VIII to the A3-C1-specific scFvs KM36 (epitope : Q1778-D1840) and KM38 (epitope : S1690-N1777 and/or V1841-N2172) with half saturation values of 84 and 165 microM, respectively, suggesting that the A3 and/or C1 domains may contribute to membrane binding of the cofactor. PMID- 15886797 TI - Elevated factor VIII is a risk factor for idiopathic venous thromboembolism in Canada - is it necessary to define a new upper reference range for factor VIII? AB - Previous studies suggest elevated factor VIII is a common, independent risk factor for venous thromboembolism (VTE); however, these studies included secondary and idiopathic VTE. We sought to explore the association between elevated factor VIII and VTE in Canadian patients with idiopathic thrombosis, and confirm the current upper factor VIII reference range was appropriate. We enrolled 300 consecutive patients with idiopathic VTE who were matched to friend controls by age, sex and ethnicity. Factor VIII levels were measured and compared between cases and controls. The optimal cut-off value to designate factor VIII levels as elevated was determined using a variety of methods. The optimal upper cut-off value for factor VIII levels was 270 IU/dl. In the logistic regression analysis, cases were more likely to have elevated factor VIII levels (OR:8.76), as were females (OR:1.93) and older subjects (OR: 1.05). Factor VIII cutoffs for a 95% specificity by age were 238 IU/dl for subjects <40 years, 248 IU/dl age 40 55 years, 261 IU/dl age 56-70 years, and 313 IU/dl age >70. Our findings confirm that elevated factor VIII is associated with an increased risk of idiopathic VTE. In our patients and matched controls, the current upper limit of normal (150 IU/dl) for factor VIII is not of clinical use. We propose that the upper limit be increased to 270 IU/dl or individual labs should establish their upper limit if they wish their assays to be discriminatory in patients with VTE. Age specific cut-offs may be clinically relevant. PMID- 15886798 TI - Does human semen contain a functional haemostatic system? A possible role for Tissue Factor Pathway Inhibitor in fertility through semen liquefaction. AB - There is already evidence that a few components of the haemostatic system exist in semen. If these comprise a functional system, they may have a role in seminal clotting and liquefaction processes and ultimately may influence fertility. What might be expected in semen as collected from fertility clinics i.e., after having both coagulated and subsequently liquefied is uncertain. It does however still contain significant amounts of Tissue Factor (TF) although its effect on semen quality remains poorly understood. The present study analyses semen for Tissue Factor Pathway Inhibitor (TFPI). Measurements were made in seminal plasma, swim up sperm and prostasomes and its relationship with conventional fertility parameters assessed. TFPI antigen levels in seminal plasma were measured in a total of 176 subjects using an Enzyme-linked immunosorbent assay (ELISA). These include sub-fertile (n=37), normally fertile (n=40), fertile sperm donor (n=34), vasectomized subjects (n=65) and in a further group defined by normality in several parameters derived from the World Health Organization (WHO) fertility criteria and termed "pooled normal semen parameters" (PNSP). For characterization studies, both TFPI activity and antigen were measured on whole semen, swim-up sperm and prostasome-rich fraction (n=5). TFPI levels were significantly higher in normal men as compared to sub-fertile (P<0.01) or vasectomized subjects (P<0.001). TFPI levels were even higher in the donor quality semen and the PNSP group. TFPI levels also correlated with semen liquefaction time, normal semen viscosity, sperm progression, percentage of motile sperm and sperm counts (density). In conclusion, the present finding substantiates the concept of an active clotting system in human semen. TFPI could regulate the activity of abundant TF, as it does elsewhere. Given a functional set of coagulation factors in semen, the TF/TFPI balance might impinge on its liquefaction and hence on global fertility. PMID- 15886799 TI - Effect of oral contraceptives on the anticoagulant activity of protein S in plasma. AB - We determined anticoagulant parameters that depend on protein S function in plasma, i.e. the APC-independent anticoagulant activity of protein S (expressed as pSR) and APC resistance determined with thrombin generation-based tests (expressed as APCsr) as well as plasma levels of total and free protein S and prothrombin in men, women not using oral contraceptives (OC), and in women using second or third generation OC. Thrombin generation in the APC resistance assays was initiated either with factor Xa (Xa-APCsr) or tissue factor (TF-APCsr). The APC-independent anticoagulant activity of protein S was highest in men (pSR=1.69) and gradually decreased from women not using OC (pSR=1.49) via women using second generation (pSR=1.35) to women using third generation OC (pSR=1.27). The pSR correlated inversely with nAPCsr determined with the tissue factor-based APC resistance test (TF-APCsr) but not with nAPCsr determined with the factor Xa based assay (Xa-APCsr). Multiple linear regression analysis in which sex, OC use, and protein S and prothrombin levels were included as independent variables and the pSR, TF-APCsr or Xa-APCsr as dependent variables indicated that plasma protein S levels poorly predict the pSR and the TF-APCsr, but are the main determinant of the Xa-APCsr. This indicates that OC use alters the expression of protein S activity. This phenomenon can be caused by differences in modulation of the activity of protein S by other plasma proteins that change during OC use or by OC-induced changes in the protein S molecule that impair its anticoagulant activity. Functional impairment of protein S as a result of hormonal influence may, at least in part, contribute to the thrombotic risk of OC users. PMID- 15886800 TI - Fatal vascular outcomes following major orthopedic surgery. AB - Major orthopedic surgery is known to be associated with potentially serious arterial and venous vascular complications, although uncertainty exists about current event rates. Using electronic databases and investigator contact, we identified randomized and cohort studies reporting overall mortality and fatal vascular events. Where possible, studies reporting high autopsy rates (>60%) were examined. Pooled incidences were calculated from eligible studies. For Autopsy studies: Pooled overall mortality and fatal pulmonary embolism for patients undergoing elective hip and knee replacement without prophylaxis could not be calculated, while with prophylaxis they were 0.44% (95% confidence interval 0.02 to 0.87%) and 0.43% (0.01 to 0.85%). For patients undergoing hip fracture surgery, the corresponding rates without prophylaxis were 15.9% (14.5 to 17.3%) and 1.9% (1.4 to 2.4%). With prophylaxis, mortality and fatal pulmonary embolismrates were 8.5% (7.3 to 9.7%) and 1.0% (0.6 to 1.5%). Among Cohort studies: Pooled overall mortality and fatal pulmonary embolism for patients undergoing elective hip and knee replacement without prophylaxis were 0.93% (0.57 to 1.29%) and 0.36% (0.14 to 0.59%). For patients receiving prophylaxis (7 to 14 days), mortality and fatal pulmonary embolism were 0.57% (0.51 to 0.62%) and 0.18% (0.14 to 0.21%). Patients undergoing hip fracture surgery receiving prophylaxis hadmortality and fatal pulmonary embolismrates of 3.2% (2.8 to 3.6%) and 0.30% (0 to 0.61%). Vascular events contributed towards approximately 50% of all deaths with similar proportions due to ischemic heart disease, cardiac failure and pulmonary embolism. In conclusion, although prophylaxis results in a reduction in overall mortality and fatal pulmonary embolism, vascular events continue to be a common cause of mortality. PMID- 15886801 TI - Lipoprotein (a) and other prothrombotic risk factors in Caucasian women with unexplained recurrent miscarriage. Results of a multicentre case-control study. AB - From 1998 to 2003, 133 Caucasian women aged 17-40 years (median 29 years) suffering from unexplained recurrent miscarriage (uRM) were consecutively enrolled. In patients and 133 age-matched healthy controls prothrombotic risk factors (factor V (FV) G1691A, factor II (FII) G20210A, MTHFR T677T, 4G/5G plasminogen activator inhibitor (PAI)-1, lipoprotein (Lp) (a), protein C (PC), protein S (PS), antithrombin (AT), antiphospholipid/anticardiolipin (APA/ACA) antibodies) as well as associated environmental conditions (smoking and obesity) were investigated. 70 (52.6%) of the patients had at least one prothrombotic risk factor compared with 26 control women (19.5%; p<0.0001). Body mass index (BMI; p=0.78) and smoking habits (p=0.44) did not differ significantly between the groups investigated. Upon univariate analysis the heterozygous FV mutation, Lp(a) > 30 mg/dL, increased APA/ACA and BMI > 25 kg/m(2) in combination with a prothrombotic risk factor were found to be significantly associated with uRM. In multivariate analysis, increased Lp(a) (odds ratio (OR): 4.7/95% confidence interval (CI): 2.0-10.7), the FV mutation (OR:3.8/CI:1.4-10.7), and increased APA/ACA (OR: 4.5/CI: 1.1-17.7) had independent associations with uRM. PMID- 15886802 TI - Patients with unstable control have a poorer dietary intake of vitamin K compared to patients with stable control of anticoagulation. AB - Evidence suggests that alterations in the dietary intake of vitamin K can affect anticoagulation response to warfarin. It is possible that a low and erratic intake of dietary vitamin K is at least partly responsible for the variable response to warfarin in patients with unstable control of anticoagulation. Twenty six patients with unstable and twenty-six with stable control of anticoagulation completed dietary records of all foods and drinks consumed on a daily basis for two consecutive weeks. The mean daily intake of vitamin K in unstable patients was considerably lower than that for stable patients during the study period (29+/-17 microg v . 76+/-40 microg). The logarithm of vitamin K intake was consistently and significantly lower in the unstable patients than the stable patients over the two week period (5.9+/-0.4 microg v. 6.9+/-0.5 microg; p<0.001; 95% CI: 0.7-1.2). Changes in vitamin K intake between weeks 1 and 2 of the study were negatively correlated with changes in International Normalised Ratio (INR) amongst the unstable patients, however this failed to reach significance (r= 0.25; p=0.22). Daily supplementation with oral vitamin K in unstable patients could lead to a more stable anticoagulation response to warfarin. PMID- 15886803 TI - Prevalence of venous thromboembolism at a teaching hospital in Okinawa, Japan. AB - Limited data suggest that Asian Americans may have a lower risk of venous thromboembolism (VTE) than Caucasians. However, the actual prevalence of VTE among Asians remains controversial, and has not been described in Japan. We studied all 131,060 patients hospitalized at a single medical centre in Japan (January 1987 - December 1999). Patients with VTE were identified through discharge diagnoses. Hospital records were reviewed for information on patient demographics, risk factors, and diagnostic modalities. VTE occurred in 0.11% of admissions (n=141, 95%CI 0.09-0.13%). Mean age (+/-SD) was 64+/-17 years, 70% were women, 91% had deep vein thrombosis, and 29% pulmonary embolism. Among hospitalized patients 50-69 years old, VTE was significantly more common among women than men (0.31% vs. 0.08%; OR 3.88; 95%CI 1.45-6.31). We found a low prevalence of VTE in Japan compared to that reported in the US. Future studies are needed to clarify the reasons for our findings. PMID- 15886804 TI - P2Y12 ADP receptor-dependent tyrosine phosphorylation of proteins of 27 and 31 kDa in thrombin-stimulated human platelets. AB - In thrombin-stimulated human platelets several proteins undergo rapid and transient changes in tyrosine phosphorylation. We demonstrate that a set of proteins of 27, 29, 31, 34, and 39 kDa is affected by released ADP and P2Y12 receptor signaling during platelet activation. AR-C69931MX, an antagonist of the Gi(2)-coupled P2Y12 ADP receptor, inhibits initial tyrosine phosphorylation of p27 and p31 and prevents subsequent dephosphorylation of p29, p34, and p39. Antagonists of the Gq-coupled P2Y1 ADP receptor have no effect. Precluding integrin alpha(IIb)beta(3) outside-in signaling with RGDS or S1197 does not affect the increase in tyrosine phosphorylation of the set of proteins but inhibits their subsequent dephosphorylation. Besides the ADP analogue 2-MeS-ADP, other platelet agonists such as collagen and the TXA(2)-mimetic U46619 also induce p27 and p31 tyrosine phosphorylation in a P2Y12 receptor-dependent manner. Tyrosine phosphorylation of p27 and p31 in response to collagen, but not thrombin, is prevented by aspirin and the TXA(2) receptor antagonist SQ29548, indicating that the effect of collagen strongly relies on TXA(2) signaling. Furthermore, epinephrine, acting via inhibitory Gz-coupled alpha(2A) adrenoceptors, bypasses the inhibitory effect of AR-C69931MX on thrombin-induced p27 and p31 tyrosine phosphorylation. Finally, we demonstrate that tyrosine phosphorylation of p27 and p31 downstream of P2Y12 receptors is due to the inhibition of adenylyl cyclase but not phosphoinositide 3-kinase (PI 3-K) activation. Elevating cAMP levels with PGI(2) or forskolin precludes thrombin induced p27 and p31 tyrosine phosphorylation. Moreover, direct inhibition of adenylyl cyclase by SQ22536 reverses the effect of AR-C69931MX. Our data indicate that the observed changes in tyrosine phosphorylation are the result of both primary Gq signaling, initiating the release of ADP, as well as subsequent P2Y12 receptor-mediated Gi coupling. PMID- 15886805 TI - Characterization of a myeloma patient with a life-threatening hemorrhagic diathesis: presence of a lambda dimer protein inhibiting shear-induced platelet aggregation by binding to the A1 domain of von Willebrand factor. AB - We have identified a patient with IgD lambda-type multiple myeloma who was characterized by a severe bleeding tendency, especially after puncture of arterial vessels. Both the bleeding time (>25 min) and activated partial thromboplastin time (APTT) were prolonged. To clarify the underlying pathogenesis, we purified the APTT-prolonging activity from the patient's serum. The purified protein was a highly negatively-charged homodimer of the lambda light chain. The lambda dimer protein (M-protein) inhibited ristocetinand high shear-induced platelet aggregation, dependent on platelet glycoprotein Ibalpha (GPIbalpha), but not epinephrine-, collagen-, ADP-, thrombin-, or botrocetin induced platelet aggregation. The lambda dimer protein inhibited the binding of platelets to immobilized or ristocetin-treated von Willebrand factor (VWF). Furthermore, a 39/34 kD fragment of VWF encompassing the A1 domain specifically bound to the immobilized lambda dimer protein in the presence of ristocetin, suggesting that the lambda dimer protein directly binds to the A1 domain of VWF. To help elucidate the binding site within the A1 domain, binding of ristocetin treated VWF to the immobilized lambda dimer protein was assayed in the presence of various anti-A1 domain monoclonal antibodies. Based on these data, we conclude that the lambda dimer protein binds to the region of the A1 domain composed of helices alpha3 and alpha4 and thus interferes with VWF-GPIbalpha interaction. The existence of a protein that inhibits high shear-induced platelet aggregation in acquired von Willebrand disease (VWD) has only rarely been reported. The results suggest that the hemostatic function in arteries with high shear force is profoundly disrupted if the binding of GPIbalpha to VWF is abrogated, supporting the relevance of shear-induced VWF interaction with GPIbalpha in the initiation of the hemostatic process. PMID- 15886806 TI - Type I Glanzmann thrombasthenia caused by an apparently silent beta3 mutation that results in aberrant splicing and reduced beta3 mRNA. AB - We report a novel genetic defect in a patient with type I Glanzmann thrombasthenia. Flow cytometry analysis revealed undetectable levels of platelet glycoproteins alphaIIb and beta3, although residual amounts of both proteins were detectable in immunoblotting analysis. Sequence analysis of reversely transcribed platelet beta3 mRNA showed a 100-base pair deletion in the 3'-boundary of exon 11, that results in a frame shift and appearance of a premature STOP codon. Analysis of the corresponding genomic DNA fragment revealed the presence of a homozygous C1815T transition in exon 11. The mutation does not change the amino acid residue but it creates an ectopic consensus splice donor site that is used preferentially, causing splicing out of part of exon 11. The parents of the proband, heterozygous for this mutation, were asymptomatic and had reduced platelet content of alphaIIbbeta3. PCR-based relative quantification of beta3 mRNA failed to detect the mutant transcript in the parents and showed a marked reduction in the patient. The results suggest that the thrombasthenic phenotype is, mainly, the result of the reduced availability of beta3-mRNA, most probably due to activation of the nonsense-mediated mRNA decay mechanism. They also show the convenience of analyzing both genomic DNA and mRNA, in order to ascertain the functional consequences of single nucleotide substitutions. PMID- 15886807 TI - A novel homozygous mutation (1619delC) in GPIIb gene associated with Glanzmann thrombasthenia, the decay of GPIIb-mRNA and the synthesis of a truncated GPIIb unable to form complex with GPIIIa. AB - The absence of agonist-induced platelet aggregation and the lack of fibrinogen receptor (GPIIb/IIIa) on the platelet surface demonstrated that the severe hemorrhagic complications of a child of Romany descent were caused by Glanzmann thrombasthenia. DNA sequencing revealed a novel homozygous deletion of a cytosine (1619delC) in the GPIIb gene causing a frameshift and predicting a novel stop codon at position 533 following 24 altered amino acids. Both parents possessed the same deletion in heterozygous form. The amount of GPIIb mRNA in the patient's platelets was 0.06% of the amount measured in control platelets. Neither GPIIb nor its truncated form could be detected in the platelets of the patient by Western blotting, while a small amount of GPIIIa was demonstrated. Quantitative flowcytometric analysis showed an elevated number of vitronectin receptors, a component of which is GPIIIa, on the patient's platelets. The surface expression of vitronectin receptor on thrombasthenic, but not on normal platelets was further increased by activation with thrombin receptor agonist peptide. BHK cells transfected with wild type GPIIIa andmutated GPIIb failed to express any mature GPIIb or pro-GPIIb. Immunoprecipitation with a polyclonal antibody recognizing both GPIIb and GPIIIa recovered a 60 kDa truncated form of GPIIb. This band was absent when immunoprecipitation was carried out with an antibody recognizing GPIIIa, suggesting that the truncated protein, lacking calf-1, calf-2 domains and major part of the thigh domain, is unable to form complex with GPIIIa. PMID- 15886808 TI - Non-invasive imaging of glycoprotein VI binding to injured arterial lesions. AB - Glycoprotein VI (GPVI) is the major platelet collagen receptor and plays a critical role in the process of thrombosis at sites of atherosclerotic lesions. This study evaluates the feasibility of radiolabeled soluble GPVI to identify injured arterial lesions. Radiolabeling was carried out using the iodogen method and resulted in the radioiodinated GPVI in radiochemical yields between 97-100%. The biodistribution of [(125)I]GPVI was determined in normal mice and demonstrated a blood clearance halftime of approximately 5.5 hours. Vascular lesions were induced in the carotid artery in wild type and ApoE(-/-)mice. Immediately after injury radioiodinated GPVI was injected intravenously. Binding of [(123)I]GPVI to carotid lesions was assessed by szintigraphic in vivo imaging. Carotid arteries were explanted for ex vivo autoradiography and histological characterization of the lesion. In vivo and ex vivo imaging revealed substantial accumulation of radioiodinated GPVI in the injured artery wall, with a ratio of lesion to control vessel of 3:1 and 7:1, respectively. Because GPVI is the critical collagen receptor that mediates platelet adhesion to vascular lesions, soluble radiolabeled GPVI may be an agent for non-invasive imaging of thrombogenic thus, vulnerable atherosclerotic plaques. PMID- 15886809 TI - Recombinant albumins containing additional peptide sequences smaller than barbourin retain the ability of barbourin-albumin to inhibit platelet aggregation. AB - The previously described fusion protein BLAH(6) (Marques JA et al.,Thromb Haemost 2001; 86: 902-8) is a recombinant protein that combines the small disintegrin barbourin with hexahistidine-tagged rabbit serumalbumin (RSA) produced in Pichia pastoris yeast. We sought to determine: (1) if BLAH(6) was immunogenic; and (2) if its barbourin domain could be productively replaced with smaller peptides. Purified BLAH(6) was injected into rabbits, and anti-barbourin antibodies were universally detected in plasma 28 days later; BLAH(6) was, however, equally effective in reducing platelet aggregation in both naive and pre-treated rabbits. Thrombocytopenia was not observed, and complexing BLAH(6) to alpha(IIb)beta(3) had no effect on antibody detection. The barbourin moiety of BLAH(6) was replaced with each of four sequences: Pep I (VCKGDWPC); PepII (VCRGDWPC); PepIII (bar bourin 41-54); and PepIV (LPSPGDWR). The corresponding fusion proteins were tested for their ability to inhibit ADP-induced platelet aggregation. PepIII LAH(6) inhibited neither rabbit nor human platelets. PepI-LAH(6) and PepIV-LAH(6) inhibited rabbit platelet aggregation as effectively as BLAH(6), but PepIV-LAH(6) did not inhibit human platelet aggregation. PepI-LAH(6) and PepIILAH(6) inhibited human platelet aggregation with IC(50)s 10- and 20-fold higher than BLAH(6). Cross-immunoprecipitation assays with human platelet lysates confirmed that all proteins and peptides interacted with the platelet integrin alpha(IIb)beta(3), but with greatly varying affinities. Our results suggest that the antiplatelet activity of BLAH(6) can be retained in albumin fusion proteins in which smaller peptides replace the barbourin domain; these proteins may be less immunogenic than BLAH(6). PMID- 15886810 TI - Contrast medium attenuates platelet activation and platelet-leukocyte cross-talk. AB - The influence of ionic and non-ionic contrast media (CM) on platelet and leukocyte activation and platelet-leukocyte crosstalk was investigated in hirudinized whole blood. The blood was incubated with and without the ionic CM ioxaglate and the nonionic CM iodixanol at 37 degrees C for 5 min, without or with stirring. Platelet and leukocyte activation and platelet-leukocyte aggregation were measured using whole blood flow cytometry. When blood samples were pre-incubated in the presence of 2%, 5%, and 10% of CM without stirring, both ioxaglate and iodixanol had little effect on unstimulated samples, but dose independently decreased 1 microM ADP-induced platelet P-selectin expression and fibrinogen binding, and thus platelet-leukocyte aggregate formation. Ioxaglate had little effect on leukocyte CD11b expression, whilst iodixanol slightly enhanced resting and N-formyl-methionyl-leucyl-phenylalanine (fMLP; 0.1 microM) stimulated leukocyte CD11b expression. Blood samples were also incubated with stirring to investigate the impact of CM (5% of ioxaglate or iodixanol) on platelet-leukocyte cross-talk. Collagen induced marked platelet activation and platelet-leukocyte aggregation, and subsequently elevated leukocyte CD11b expression. The latter was attenuated by ioxaglate and iodixanol, and was accompanied by reduced platelet-leukocyte aggregation. In conclusion, the CM ioxaglate and iodixanol attenuate platelet activation and platelet-leukocyte cross-talk. Inhibitory effects of the contrast agents on this cross-talk are apparently exerted by reducing heterotypic conjugation, and may be beneficial in connection with PCI. PMID- 15886811 TI - Extracellular fibrinogen binding protein, Efb, from Staphylococcus aureus as an antiplatelet agent in vivo. AB - Staphylococcus aureus produces and secretes a protein, extracellular fibrinogen binding protein (Efb), which contributes to virulence in wound infection. We have previously shown that Efb is a potent inhibitor of platelet function in vitro. We confirm here that this is also the case in vivo. Pre-treatment with Efb resulted in a significant prolongation of bleeding time in a mouse model. Furthermore, Efb was capable of rescuing animals from death caused by the administration of potent platelet agonists. This antiplatelet effect may explain the retardation of wound healing associated with Efb in S. aureus wound infections. These results are important not only in terms of understanding S. aureus pathogenesis, and consequently identifying new treatment strategies, but also with regard to the development of potential, novel antiplatelet agents for the prevention of thrombosis. PMID- 15886812 TI - Streptococcus sanguis-induced platelet activation involves two waves of tyrosine phosphorylation mediated by FcgammaRIIA and alphaIIbbeta3. AB - The low-affinity IgG receptor, FcgammaRIIA, has been implicated in Streptococcus sanguis-induced platelet aggregation. Therefore, it is likely that signal transduction is at least partly mediated by FcgammaRIIA activation and a tyrosine kinase-dependent pathway. In this study the signal transduction mechanisms associated with platelet activation in response to the oral bacterium, S. sanguis were characterised. In the presence of IgG, S. sanguis strain 2017-78 caused the tyrosine phosphorylation of FcgammaRIIA 30s following stimulation, which led to the phosphorylation of Syk, LAT, and PLCgamma2. These early events were dependent on Src family kinases but independent of either TxA(2) or the engagement of the alpha(IIb)beta(3) integrin. During the lag phase prior to platelet aggregation, FcgammaRIIA, Syk, LAT, and PLCgamma2 were each dephosphorylated, but were re phosphorylated as aggregation occurred. Platelet stimulation by 2017-78 also induced the tyrosine phosphorylation of PECAM-1, an ITIM-containing receptor that recruits protein tyrosine phosphatases. PECAM-1 co-precipitated with the protein tyrosine phosphatase SHP-1 in the lag phase. SHP-1 was also maximally tyrosine phosphorylated during this phase, suggesting a possible role for SHP-1 in the observed dephosphorylation events. As aggregation occurred, SHP-1 was dephosphorylated, while FcgammaRIIA, Syk, LAT, and PLCgamma2 were rephosphorylated in an RGDS-sensitive, and therefore alpha(IIb)beta(3)-dependent, manner. Additionally, TxA(2) release, 5-hydroxytryptamine secretion and phosphatidic acid formation were all blocked by RGDS. Aspirin also abolished these events, but only partially inhibited alpha(IIb)beta(3) -mediated re phosphorylation. Therefore, S. sanguis -bound IgG cross links FcgammaRIIA and initiates a signaling pathway that is down-regulated by PECAM-1-bound SHP-1. Subsequent engagement of alpha(IIb)beta(3) leads to SHP-1 dephosphorylation permiting a second wave of signaling leading to TxA(2) release and consequent platelet aggregation. PMID- 15886813 TI - A potential role of YC-1 on the inhibition of cytokine release in peripheral blood mononuclear leukocytes and endotoxemic mouse models. AB - To evaluate the anti-sepsis potential of YC-1, we have examined the effect of YC 1 on the regulation of cytokine production in human leukocytes and endotoxemic mice. The data demonstrated that YC-1 showed a preferential inhibition on proinflammatory cytokine production without inhibition of cell growth or induction of cytotoxicity in human leukocytes. On the other hand, in the septic mouse model, treatment with an intraperitoneal application of LPS caused a cumulative death within 27 hours. The post-treatment administration of YC-1 significantly increased the survival rate in endotoxemic mice. Furthermore, several mediators were detected and the data showed that YC-1 profoundly blocked LPS-induced NO as well as TNF-alpha production, and prevented lung damage by histological examination. Samples from the animal model showed that LPS-induced NF-kappaB/DNA binding activity and consequent up-regulation of iNOS expression in tissues were abolished by post-administration of YC-1. Furthermore, YC-1, by itself, did not modify cGMP content while significantly inhibit LPS-induced cGMP formation, suggesting that YC-1-mediated effect was not through a cGMP-elevating pathway. Taken together, it is evident that the post-treatment administration of YC-1 after LPS application significantly inhibits NF-kappaB activation, iNOS expression, NO over-production, and cytokine release reaction resulting in an improved survival rate in endotoxemic mice. It is suggested that YC-1 may be a potential agent for the therapeutic treatment of sepsis. PMID- 15886814 TI - Opposite effects of CX3CR1 receptor polymorphisms V249I and T280M on the development of acute coronary syndrome. A possible implication of fractalkine in inflammatory activation. AB - Several lines of evidence suggest that the chemokine fractalkine (FKN) and its receptor CX3CR1 contribute to the accumulation of leukocytes in the atherosclerotic plaque. The M280 allele of the CX3CR1T280M polymorphism modulates leukocyte recruitment and is associated with lower prevalence of cardiovascular disease. The influence of V249I, another CX3CR1 polymorphism, is discussed controversially. We investigated the association of the alleles M280 and I249 of CX3CR1 with coronary artery disease (CAD) and with acute coronary syndrome (ACS). Additionally, we assessed their association with the soluble ligand FKN and inflammatory activation measured by high sensitivity C-reactive protein (hsCRP). The genotypes of the V249I and T280M polymorphisms were determined in 1152 patients with suspected CAD.720 (62.5%) individuals showed significant CAD with an ACS prevalence of 59.3%. Using multivariate regression, we found a harmful influence of I249 (adjusted OR=1.8, P<0.03) and a protective effect of M280 (adjusted OR=0.6, P<0.04) on the occurrence of ACS in patients with CAD. Correspondingly, patients with I249 but without M280 (17%) were at elevated risk of ACS (OR=1.6, P<0.04). During ACS these patients (carrying only I249) had significantly higher circulating concentrations of FKN and high sensitivity C reactive protein (1.9- and 1.6-fold). We found no association of the I249 or the M280 allele with the occurrence of CAD. In conclusion, I249 and M280 have opposite effects on the occurrence of ACS. The presence of I249 not "balanced" by M280 confers an elevated risk of ACS. A FKN-mediated enhanced inflammatory activation might explain this increased risk. PMID- 15886815 TI - Associations of fibrinogen and C-reactive protein with prevalent and incident coronary heart disease are attenuated by adjustment for confounding factors. British Women's Heart and Health Study. AB - A cross sectional and prospective analysis of 3,745 British women aged 60-79 years at baseline was undertaken. Among these women there were 570 prevalent cases of coronary heart disease (CHD) and 151 new cases among 12,641 person-years of follow up of women who were free of CHD at baseline. Both fibrinogen and CRP were associated with indicators of socioeconomic position in childhood and adulthood and there was a cumulative effect of socioeconomic position from across the life course. The age-adjusted odds ratio (95% confidence interval) of prevalent CHD for a 1 unit (1 g/L) increase in fibrinogen was 1.29 (1.12, 1.49); with full adjustment for all potential confounding factors this attenuated to 1.09 (0.93, 1.28). The hazards ratio for incident CHD among those free of disease at baseline was 1.28 (1.00, 1.64); with full adjustment for all potential confounding factors this attenuated to 1.09 (0.84, 1.44). Similar effects of adjustment for confounding factors were seen for the associations between CRP and both prevalent and incident CHD. By contrast, the strong positive association between smoking (an established causal risk factor for CHD) and CHD was not attenuated by adjustment for life course socioeconomic position or other risk factors. We conclude that fibrinogen and CRP predict CHD but may not be causally related to it. PMID- 15886816 TI - Epidermal growth factor modulates prostate cancer cell invasiveness regulating urokinase-type plasminogen activator activity. EGF-receptor inhibition may prevent tumor cell dissemination. AB - Urokinase-type plasminogen activator receptor (uPAR) and Epidermal Growth Factor Receptor (EGFR) are ubiquitous receptors involved in the control of a variety of cellular processes frequently found altered in cancer cells. The EGFR has been recently described to play a transduction role of uPAR stimuli, mediating uPA induced proliferation in highly malignant cells that overexpress uPAR. We compared the uPA production, the presence of uPAR, AR, EGFR and Her2 with the chemotaxis and the Matrigel invasion in ten human PCa cell lines and observed that: (1) the levels of Her2, but not of EGFR, as well as the uPA secretion, cell motility and Matrigel invasion were statistically higher in AR negative than in AR positive PCa cells; (2) the uPA secretion and uPA Rexpression were positively related to Matrigel invasion; (3) the EGF was able to stimulate chemotaxis and Matrigel invasion in a dose-dependent manner; (4) the EGF-induced cell migration was statistically higher inAR negative than in AR positive cells with a similar increase with respect to basal value (about 2.6 fold); (5) the Matrigel invasion was statistically higher in AR negative than in AR positive PCa cells also if the increment of Matrigel invasion after EGF treatment was statistically higher in AR positive respect to AR negative cells; (6) the EGF induced uPA secretion and its membrane uptake through the increment of uPAR; and (7) these effects were blocked by EGFR/Her2 tyrosine kinase inhibitors with IC(50) lower than those needed to inhibit cell proliferation and required PI3K/Akt, MAPK and PI-PLC activities as verified by inhibition experiments. These enzymatic activities were regulated in different manners in PTEN positive and negative cells. In fact, the inhibition of PI3K blocked the EGF-induced invasiveness in PTEN positive cells but not in PTEN negative cells, in which PI3K activity was not influenced by EGFR/Her2 activation, whereas the inhibition of MAPK was able to block the invasive phenomena in both cell types. Taken together, our data suggest that the blockade of EGFR could attenuate the invasive potential of PCa cells. In addition, considering that the EGFR expression is related to higher Gleason grade of PCa and that EGFR levels are increased after anti androgenic therapy, this therapeutic approach could slow down the metastasis formation which represents the most dramatic event of PCa progression. PMID- 15886817 TI - Von Willebrand gene tracking by single-tube automated fluorescent analysis of four short tandem repeat polymorphisms. AB - Molecular diagnosis of von Willebrand disease (VWD) has been hampered by the large size and complex genomic characteristics of the gene involved. For this reason, indirect methods using intragenic polymorphic markers described along the von Willebrand factor (VWF) gene are valuable tools for gene monitoring and linkage analysis. Several studies have demonstrated the four commonly utilized short tandem repeats (STRs), three located in intron 40 and one in the promoter region of the VWF gene, to be highly informative for this task. Our objective was t o develop a rapid, automated method to simultaneously analyze these four STRs for VWF gene tracking. Amplification of the four loci is achieved in a single multiplex fluorescent PCR which is then analyzed in the same run by capillary electrophoresis. Data processing with GeneScan and Genotyper software has simplified management and tabulation of the resulting haplotypes. Analysis of the VWF gene in DNA from 102 individuals (204 chromosomes) revealed that the three STRs within intron 40 showed significant linkage disequilibrium against each other but not against the VWP locus. Moreover, the combination of the four markers offers a high heterozygosity rate (>99%) that improves tracing VWF gene inheritance. In conclusion, the automated fluorescent capillary electrophoresis method presented here is an extremely rapid, simple and highly informative technique for association studies between VWD and the VWF gene in addition to genetic counseling and prenatal diagnosis by precise linkage analysis in VWD affected families. PMID- 15886818 TI - Diagnostic management of pulmonary embolism using clinical assessment, plasma D dimer assay, complete lower limb venous ultrasound and helical computed tomography of pulmonary arteries. A multicentre clinical outcome study. AB - The objective of the study was to assess the clinical validity of a non-invasive diagnostic strategy for acute pulmonary embolism using clinical assessment combined with both ELISA D-dimer and complete lower limb ultrasound (US) examination of proximal and distal veins, before single-detector helical computed tomography (CT) of pulmonary arteries. We expected the strategy to have a high diagnostic exclusion power and to safely decrease the number of CT scans. This prospective, multicenter outcome study included 274 consecutive outpatients. All underwent a priori clinical probability, D-dimer and bilateral complete lower limb US assessments. Only patients with a high clinical probability and both tests negative, or positive D-dimer and negative US assessments, underwent CT. This was deemed necessary in 114 patients (42%). At baseline, venous thromboembolism (VTE) was detected in 110 patients (40%), either by US showing proximal (n=65) or distal (n=36) thrombosis, or by CT (n=9). Anticoagulant was withheld in the remaining patients with negative results in both D-dimer and US but a non-high clinical probability (n=59), or in both US and CT (n=90), or with negative US (n=6) and inadequate CT (n=9). All patients underwent a three-month clinical follow-up. VTE occurred in one patient with inadequate CT, yielding an incidence of 0.6% [95% confidence interval: 0.1-3.4]. No patient died from VTE or had major bleeding. Using clinical probability, ELISA D-dimer and complete US before helical CT is a safe strategy resulting in a substantial reduction in CT scans. PMID- 15886819 TI - Accelerated exposure of phosphatidylserine on lymphocyte populations from patients with systemic lupus erythematosus or rheumatoid arthritis. PMID- 15886820 TI - Value of ventilation/perfusion SPECT detecting extensive pulmonary embolism in a patient with pneumonia. PMID- 15886821 TI - A child with multifocal arterial thrombosis. PMID- 15886822 TI - Complexity of the genetic contribution of different thrombotic risk factors in a Spanish thrombophilic family. PMID- 15886823 TI - Successful treatment of heparin induced thrombocytopenia (HIT) with fondaparinux. PMID- 15886824 TI - Rituximab in the treatment of factor XIII inhibitor possibly caused by Ciprofloxacin. PMID- 15886825 TI - [Try to improve journal quality by improving standards and editing process]. AB - The quality of medical journals depends on several factors involving 3 groups of people with their independent but relevant roles: the authors, the reviewers and the editors. Peer review and editing is the key factor to improve the quality of medical publications and journals. Shanghai Journal of Stomatology (SJS) has been regarded as a leading journal for publishing high-quality work in the field of stomatology in China. In October 2003, it was accepted by the National Library of Medicine, USA, to be indexed and included in Index Medicus and MEDLINE. To further improve the journal's overall quality, the Editorial Agency led by Professor Zheng Jia-wei has made great efforts to formulate its essential requirements in paper style, bilingual abstract writing and statistical analysis for the manuscripts submitted for possible publication. Strict independent peer review system has been adopted to assess the quality of the manuscripts received since it was founded in 1992. The reviewer is required to address detailed aspects of the paper under review and to resend his or her opinion on the paper. The editorial management is a crucial part of the publishing process. The editors begin action with the receipt of the manuscript, direct the various steps of evaluation, correction and re-submission, until a decision is made to accept or reject the paper at the regular meeting of Decision Making Group on Manuscripts of SJS led by the Editor-in-Chief. Once a paper is accepted and carefully revised, the editors will make necessary text and layout editing. Due consideration is given to the statistical, bilingual and ethical aspects as well as to the overall uniformity of the terminology, nomenclatures and style throughout the volume as a whole in the promotion of standards. The journal has not been cited by Science Citation Index (SCI) till now, further steps should be taken to make this journal better known throughout the World, to improve the quality of the publications and reduce the delay between the initial receipt and the final publication of manuscripts. PMID- 15886826 TI - [Tooth tissue engineering: from cells to organ, an odyssey far from finished]. AB - Tooth tissue engineering is an emerging biotechnique that will provide replacemental teeth for patients suffering from different diseases causing tooth loss. Although some attempts have been tried to generate whole tooth both in vivo and in vitro, the lack of the knowledge for tooth initiation and development, as well as for tooth shape controlling mechanisms greatly impede the progress of this technique. This article reviewed and discussed some recent findings in tooth tissue engineering related to the cell resource, the concept of reconstruction and regeneration, the application of artificial scaffolds, together with the methods of organ culture and implantation. PMID- 15886827 TI - [ITI dental implants immediately placed into vascularized iliac bone grafts for reconstruction of mandibular defect: analysis of 12 consecutive cases]. AB - PURPOSE: Vascularized iliac bone graft combined implantation has become a routine procedure in the functional reconstruction of mandibular defects. The aim of this study was to assess the clinical outcomes of the implants immediately placed into the vascularized iliac bone grafts. METHODS: In this study, 12 consecutive patients with 36 ITI dental implants who had reconstruction of mandibular defect between 2000 and 2004 were presented. All implants were inserted immediately after bone grafting and loaded after 3 to 5 months of submerged healing. RESULTS: In general, a primary stability for implants placed in vascularized iliac bone grafts were achieved. After 1 year of observation the mean vertical bone loss was less than 1mm. There was no implant failure in the observation period. CONCLUSION: The vascularized iliac bone grafts are safe implant-bearing areas and allows a reliable and predictable restoration with dental implants. Implant supported bridges and overdentures can be used in this procedure. PMID- 15886828 TI - [Clinical analysis of 13 cases of hemangioma and vascular malformation associated with thrombopenia]. AB - PURPOSE: To explore the diagnosis and treatment of hemangioma and vascular malformation associated with thrombopenia (Kasabach-Merritt syndrome, KMS). METHODS: From October 1997 to December 2003, 13 cases of KMS were treated in our hospital. Among the 13 patients, 4 were located in the maxillofacial region, 3 were located in the trunk, 6 were located in the lower limb. The size of the lesion of all patients exceeded 8 cm; 10 were hemangioma, 1 was arteriovenous malformation (AVM), 1 was venous malformation (VM), 1 was Klippel-Trenaunay Syndrome (KTS). The platelet count was all lower than 70 x 10(9)/L, the lowest was 10 x 10(9)/L, the average was 41 x 10(9)/L. The clinical characteristics and course of treatment were analyzed. RESULTS: 9 patients were cured, 1 improved, 1 had no response, and 2 died. The treatment of choice for KMS was steroids, but the response rate was not high (23.08% in this series). If patients had no response to steroids, they also had no response to interferon. CONCLUSIONS: If a proper treatment was taken in early stage, most patients could get a good result, but for patients with an extensive vascular malformation that can't be removed, the prognosis was poor. For lesions in the limbs and trunk, pneumatic compression therapy has certain curative effect, fewer side effects, therefore worthy of popularization. PMID- 15886829 TI - [Application of temporal anterior hairline incision in internal fixation of zygomatic complex fracture]. AB - PURPOSE: To investigate the methods, merits and effects of temporal anterior hairline incision in internal fixation of zygomatic complex fracture (ZCF). METHODS: 99 cases of ZCF were treated via vertical incision in temporal anterior hairline. During dissection, the zygomatic branch and the frontal branch of the facial nerve, which crossed over the zygomatic arch, were protected. The deep fascia superficial to the zygomatic arch was horizontally incised, the periosteum was elevated at the fracture sites. Cases of type A1 were treated through the above approach; while minor periorbital and oral vestibule incisions were also used in cases of type B and C. The fracture segments were reduced accurately and fixed with titanium microplates. Coronal incision was not applied in all cases. RESULTS: The clinical results were good in 99 cases (86.0%). All the temporal anterior hairline incisions healed primarily. The rate of temporary facial paralysis, due to injury of the zygomatic and frontal branch of the facial nerve, was 24.3%; but all recovered six months later, without any permanent paralysis. CONCLUSION: The temporal anterior hairline incision has many merits including clear field of operation, convenient manipulation, minimal trauma, less bleeding, high accuracy of reduction and inconspicuous scar. PMID- 15886830 TI - [Reconstruction of total lower lip or corner of mouth defects with forearm free flap suspending over fascia: report of 10 cases]. AB - PURPOSE: To improve the contour and function of the lip and its commissure with forearm free flap suspending over fascia. METHODS: Since 2001,this method was used in 10 cases. Forearm free flap was designed according to the lip or buccal defects, then the defects were reconstructed with the flap through vascular anastomosis. The skin island for reconstruction of the intraoral lining and the skin defect was folded over the palmaris longus tendon or fascia lata. RESULTS: All the patients were followed-up for 2 years, the survival rate of the free flap was 100%, 2 patients had oral incompetence, the others had good oral function and sufficient oral competence. The flaps were helpful to expression of motion. CONCLUSION: Reconstruction of total lower lip or corner of mouth defects with forearm free flap suspending over fascia was helpful for rehabilitation of contour and oral function. PMID- 15886831 TI - [Application of Branemark system's multi-unit abutment: report of 37 consecutive cases]. AB - PURPOSE: To evaluate the short-term clinical effect and advantages of Branemark system's multi-unit abutment used for standard or MK III implants supported fixed prosthodontics. METHODS: Routine clinical examinations and preparations, including panoramic tomography, periapical radiograph and surgical guide plate, were performed in 37 cases with multiple lost teeth. A total of 117 Branemark system's implants were placed using a two-stage surgical approach. Multi-unit abutment connection was performed 3-6 months after implant installation. All superstructure prosthetic appliances were porcelain-fused-to-golden metal bridges. RESULTS: The follow-up period for the implants was 12 to 24 months. The total survival rate was 95.7%. Two implants were lost at second-stage surgery (the survival rate was 98.29% for first-stage), and 3 implants were lost after loading (the survival rate was 97.43% for second-stage). The other 112 implants function uneventfully. There were no bone loss around implants, no abutment and gold cylinder screw loosen. CONCLUSIONS: The multi-unit abutment, on basis of collecting all the merits of the traditional abutment, was further designed in a simplified way, which not only expands its clinical application, operate easily, but also enhance its whole superstructure. It is more suitable for implant supported fixed prosthodontics with high success rate and more advantages than the traditional abutment. PMID- 15886832 TI - [A serious complication-obstructive sleep apnea hypopnea syndrome after velopharyngeal ring ligation procedure: report of 6 cases]. AB - PURPOSE: To discuss the occurrence and characteristics of obstructive sleep apnea hypopnea syndrome (OSAHS) after velopharyngeal ring ligation procedure (VRLP). METHODS: Seven hundred and eight cleft palate patients underwent velopharyngeal ring ligation procedure from 1997 to 2003. Nine cases were found having some suspicious symptoms of OSAHS after operation and 6 of them were confirmed as OSAHS by polysomnography. Six VRLP patients without OSAHS as controls were examined with polysomnography. Chi-square test and t test were used to determine the statistical differences between the groups. RESULTS: Six cases of 708 patients met the criterion of severe OSAHS and the incidence of OSAHS after VRLP in children below 12 years was 1.8%. Compared with controls, more arousals and longer awakening were found in OSAHS group, the ratio of III and IV phase of NREM and REM in total sleep was significantly decreased (P<0.01). CONCLUSION: OSAHS was likely to occur in VRLP patients below 12 years. Many feature of this kind of OSAHS were different from the ordinary pediatric OSAHS. It showed that OSAHS after VRLP was characterized by more apneas, less partial upper airway obstruction and associated with distinguished staged desaturation. PMID- 15886833 TI - [CT evaluation of malignant tumors of upper cervical lymph nodes invading the carotid sheath]. AB - PURPOSE: The aim of this article is to evaluate the CT findings of malignant tumors of upper cervical lymph nodes invading the carotid sheath. METHODS: Seventy-seven patients (83 lesions) with malignant tumors in the upper cervical lymph nodes shown on CT scans were collected. All lesions were certificated with pathological examination by operation or biopsy. RESULTS: The CT manifestations of malignant tumors of upper cervical lymph nodes invading the carotid sheath included :(1) compression and deformation of vessels in the carotid sheath in 52 lesions (ICA, one lesions; ICV, 51 lesions); (2) disappearance of vessels in the carotid sheath (ICV, 13 lesions); (3) displacement of vessels of the carotid sheath in 67 lesions (ICA, 22 lesions; ICV, 50 lesions); and (4) malignant tumors encompassing the carotid sheath in 15 lesions. CONCLUSIONS: On CT images, the most common form of malignant lymph node lesions invading the carotid sheath is displacement of ICA and ICV. It is concluded that all forms of affecting carotid sheath demonstrating on the CT images are important in diagnosing the malignant invasion of the ICA and ICV. PMID- 15886834 TI - [Changes of circulating IFN-gamma, IL-4 in patients with chronic periodontitis before and after periodontal initial therapy]. AB - PURPOSE: To investigate the effect of periodontal initial therapy on circulating blood serum interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in patients with chronic periodontitis. METHODS: 15 patients with chronic periodontitis and 15 healthy people were selected. Circulating blood serum IFN-gamma and IL-4 were measured using double antibody sandwich ELISA; Student's t test was used for comparing sulcus bleeding index (SBI), probing depth (PD), and circulating IFN gamma, IL-4 in patients with chronic periodontitis that were assessed respectively before and 4 to 6 weeks after periodontal initial therapy. RESULTS: The level of blood serum IFN-gamma, IL-4 in patients with chronic periodontitis was significantly higher than the control group (P<0.01); After periodontal initial therapy, SBI, PD, and circulating IFN-gamma concentration were reduced significantly (P<0.01); IL-4 concentration had no significant difference (P>0.05). CONCLUSIONS: Blood serum IFN-gamma, IL-4 were associated with chronic periodontitis; The control of gingival infection may reduce blood serum IFN-gamma concentration; IL-4 concentration was not reduced with the control of gingival infection. Periodontal initial therapy. PMID- 15886835 TI - [Comparison of dentomaxillary pantomography and periapical radiographs with horizontal tube shift in localizing the impacted teeth]. AB - PURPOSE: To compare the clinic value between dentomaxillary pantomography and periapical radiographs in localization of the impacted teeth. METHODS: 43 impacted teeth were localized with both dentomaxillary pantomography technique and periapical radiographs with horizontal tube shift which is clinically widely used. And a comparison between the two methods was carried out using Chi square test. RESULTS: Both dentomaxillary pantomography and periapical radiographs with horizontal tube shift can relatively precisely demonstrate the position of the impacted teeth. The percentage of the cases which the image and the result of surgery was consistent in the two methods was 93.02% and 95.35% (P>0.05) respectively. There was no statistical difference between the two groups. CONCLUSIONS: Dentomaxillary pantomography can precisely localize the impacted teeth. PMID- 15886836 TI - [Clinical study of the effect of preventing dentine hypersensitiveness by using Fluor Protector and Green Or on the prepared vital pulp abutment teeth]. AB - PURPOSE: To study and evaluate the effect preventing dentine hypersensitiveness by using Fluor Protector or Green Or on the prepared vital pulp abutment teeth of PFM bridges. METHODS: 118 cases, 246 prepared vital pulp abutment teeth, were randomly divided into three groups: Experimental Group A--treated with the Fluor Protector and temporary crown; Experimental Group B--treated with the Green Or and temporary crown, and Control Group--only using temporary crown. The results of desensitization in 3 groups were evaluated. F test was used for analysis (DSPV6.01). RESULTS: Significant differences were found between experimental Group A, B and the control group after 1 week (when cementing the PFM bridges); and also after 1 month (P<0.05). But no significant difference was found between experimental Group A and B (P>0.05). CONCLUSIONS: The effect of preventing dental hypersensitiveness by using Fluor Protector or Green Or on the prepared vital pulp abutment teeth of PFM bridges is ideal. It is easy to use and worth being widely applied. PMID- 15886837 TI - [The effect of regular scaling on periodontal health in fixed orthodontic treatment]. AB - PURPOSE: The purpose of this study was to investigate the effect of scaling and teeth brushing with YAHAO toothpaste (eugenol and borax cream) on periodontal health in fixed orthodontic treatment. METHODS: 48 patients who needed fixed orthodontics were chosen to enroll a single-blind randomized study. The patients were randomly divided into two groups: the control group and the treatment group. Patients in the treatment group were treated with scaling and teeth brushing with YAHAO toothpaste. The changes of GI, PLI and SBI were recorded at the beginning of the treatment and after 3 months. SPSS 10.0 software package was used for Student's t test and Chi-square test. RESULTS: There was no significant difference in the periodontal indexes between the two groups before treatment. But there was a statistically significant difference in the incidence of gingivitis (P<0.01) between the two groups after treatment. CONCLUSION: The increase in the periodontal index and the rate of gingivitis in the treatment group were statistically significantly lower than that in the control group, which means that using scaling and teeth brushing with YAHAO toothpaste can effectively reduce the rate of gingivitis. PMID- 15886838 TI - [Role of Smad signaling in transcription of Smad7 gene mediated by TGF-beta1 in odontoblast cell line MDPC-23]. AB - PURPOSE: To investigate the role of Smad signaling in transcription of Smad7 gene mediated by TGF-beta1 in odontoblast cell line MDPC-23, and to explore the molecular mechanism of Smad7 gene expression mediated by TGF-beta1 at the transcriptional level. METHODS: Smad function and its role in transcription of Smad7 were investigated in cotransfection experiments using Smad7 promoter luciferase reporter construct containing the sequence between -408 bp and +112 bp of mouse Smad7 gene. The data were analysed by one-way ANOVA. RESULTS: When the Smad7 promoter-luciferase reporter gene construct was expressed in MDPC-23 cells, its transcriptional activity was significantly induced by TGF-beta1 treatment, whereas not by BMP-2 treatment. Overexpression of Smad1, 2, 4, or 5 had no effect on transcriptional activity of Smad7 promoter. Overexpression of Smad3 markedly promoted transcriptional activity of Smad7 promoter, whereas co-transfection of Smad3 and Smad4 doubled the effect of Smad3. Overexpression of Smad3 dominant negative mutant or Smad3 antisense cDNA (AS-Smad3) significantly inhibited transcriptional activity of Smad7 promoter induced by TGF-beta1. CONCLUSION: TGF beta1 regulated transcription of Smad7 gene through association of Smad3 and Smad4 in MDPC-23 cells. PMID- 15886839 TI - [Influence of storage methods on microtensile bond strength of dentin adhesive system]. AB - PURPOSE: To compare the effects of teeth stored with different methods and different durations of storage on the microtensile bond strength of the dentin adhesive system Single Bond. METHODS: 30 human first premolars were stored immediately after extraction in one of five commonly used methods respectively: 0.02% distilled water and thymol, 10% formalin, 1% chloramine, distilled water at 4 degrees C and were refrigerated at -20 degrees C. After 10 days, 90 days, the dentin adhesive system Single Bond and composite Z250 were applied to superficial occlusal flat dentin according to the manufacturers' instructions and microtensile bond strength measurements were evaluated, freshly extracted teeth being used as control. Stero-microscope and scanning electronic microscope were used to evaluate the fracture modes of the microtensile bond strength specimens. Two-way ANOVA was used for analysis of the microtensile bond strengths among different groups. RESULTS: The results showed that there was significant difference among five tooth storage methods on the microtensile bond strength of dentin bonding agent (P=0.01). Compared with freshly extracted teeth, two of five storage methods/media, 0.02% distilled water and thymol at 4 degrees C (P=0.008) and distilled water at 4 degrees C (P=0.024), resulted in significantly lower microtensile bond strengths. The duration of teeth storage had no effect on the microtensile bond strength of dentin bonding agent (P=0.279). The interaction of two factors was significant (P=0.000). Stero-microscope and SEM examination indicated that all fracture modes were adhesive failures. CONCLUSIONS: From this study, it can be concluded that the teeth storage methods/media can influence the microtensile bond strength of Single Bond adhesive system. If sufficient numbers of freshly extracted teeth are not available for bond strength test, the freezing teeth at -20 degrees C and teeth stored in 1% chloramine at 4 degrees C are preferred . PMID- 15886840 TI - [Generation of dspp-LacZ transgenic mouse founders]. AB - PURPOSE: To establish a transgenic mouse founders in which the expression of LacZ was directed by a dentin sialophosphoprotein-specific promoter. METHODS: The DSPP specific promoter was obtained by PCR and confirmed by sequencing, and the transgenic plasmid, pTN-DPM-LacZ, was constructed by subcloning the DSPP-specific promoter and LacZ-encoding sequence into one vector. The linearized transgenic plasmid was microinjected into the male pronucleus of the zygotes, and the microinjected zygotes were implanted to recipient pseudopregnant mice. The tail DNA of 4 week-pups was tested by PCR. RESULTS: 503 embryos were implanted to 20 recipient pseudopregnant mice, 12 of the 89 pups carrying the transgene. The establishment of the dspp-LacZ transgenic mouse line is under progress. CONCLUSION: 12 founders of the dspp-LacZ transgenic mice, in which the expression profile of LacZ should be the same as that of DSPP, were obtained by microinjection successfully, and the mouse line which is being established could be used as a good tool to investigate the exact expression profile of DSPP in the future. PMID- 15886841 TI - [The expression of Fas and bcl-2 in hamster buccal carcinogenesis]. AB - PURPOSE: To study the expression of Fas and bcl-2 in oral carcinogenesis. METHODS: Using immunohistochemical method,we studied the expression of Fas and bcl-2 in DMBA induced hamster baccal carcinogenesis including negative control (14), hyperkeratosis (7), epithelial dysplasia (mild 12, moderate 7, severe 9), oral squamous cell carcinoma(OSCC) (25). Non-parametric test was used for statistical analysis. RESULTS: Fas was expressed in negative control consistently except stratum basale,and there was down-regulation in oral dysplasia and OSCC (chi(2)=45.576, P<0.05).bcl-2 was rarely expressed in negative control, and there was up-regulation in oral dysplasia and OSCC (chi(2)=19.433, P<0.05). CONCLUSIONS: Fas and bcl-2 may play an important role in inhibiting apoptosis coordinately, resulting in the imbalance of cell proliferation and apoptosis, which may contribute to oral carcinogenesis. PMID- 15886842 TI - [The expression of PTEN and PKB in salivary adenoid cystic carcinoma and its clinical significance]. AB - PURPOSE: To investigate the expression of PTEN and PKB in salivary adenoid cystic carcinoma (SACC) and its clinical significance. METHODS: The expression of PTEN and PKB in 63 cases SACC were evaluated by SP immunohistochemistry. SPSS 11.5 software was used for chi-square test. RESULTS: In 38 cases, both non-cancer tissue near by cancer and cancer tissue, 21 (55.26%) cases cancer tissue PTEN expression positive were found, 32 (84.21%) cases the non-cancer tissue near by cancer PTEN expression positive were found. There was significant difference in cancer tissue and non-cancer tissue near by cancer (P<0.05). 31 (81.58%) cases cancer tissue PKB expression positive were found, 20 (52.63%) cases the non cancer tissue near by cancer PKB expression positive were found. There was significant difference in cancer tissue and non-cancer tissue near by cancer (P<0.01). In 25 cases with metastasis, 9 (36%) cases PTEN expression positive were found; in 38 cases with non-metastasis, 26 (68.42%) cases PTEN expression positive were found; there was significant difference in cases of metastasis and non-metastasis (P<0.05). In 25 cases with metastasis, 24 (96%) cases PKB expression positive were found; in 38 cases with non-metastasis, 27 (71.05%) cases PKB expression positive were found; There was significant difference in cases of metastasis and non-metastasis (P<0.05). PTEN expression and PKB expression were no significant difference between the different sex, age and sorts of pathology. PTEN and PKB were negatively correlated (P<0.01). CONCLUSIONS: The expression of PTEN and PKB may be related to the occurrence and progress of salivary adenoid cystic carcinoma,PTEN and PKB may have effect on metastasis. PMID- 15886843 TI - [A study of frictional resistance of archwires and ligating methods]. AB - PURPOSE: To study the effect of 4 archwires and 2 ligating methods on the frictional resistance. METHODS: The static and dynamic friction of different combinations of 4 archwires and 6 preadjusted brackets and 2 ligating methods in the buccal segments were tested in the dry state. The friction was tested by the load cell in the LJ-500 testing machine. Orthogonal experiment design was performed in this study. The data were analysed by analysis of variance and regression analysis using the statistical analysis system. RESULTS: The smallest frictional resistance in 4 different archwires was produced by combination of 0.018 x 0.025 inch stainless rectangular wire and all preadjusted brackets. The biggest frictional resistance in 4 different archwires was produced by combination of 0.019 x 0.025 inch stainless rectangular wire and all preadjusted brackets. The static friction of combination of 0.020 inch stainless round wire and all preadjusted brackets were higher than 0.018 inch stainless round wire. The dynamic friction of combination of 0.018 inch stainless round wire and all preadjusted brackets were higher than 0.020 inch stainless round wire. Among 4 kinds of stainless wires, the ratio of dynamic to static friction of 0.018 inch round wire appeared the highest, followed by 0.018 x 0.025 inch rectangular wire, 0.020 inch round wire and 0.019 x 0.025 inch rectangular wire. Elastomeric ring ligature produced higher friction and the ratio of dynamic to static friction than stainless steel ligature. CONCLUSIONS: 0.018 inch stainless steel round wire was not suitable for sliding mechanics in this study. One should pay attention to anchorage control in using 0.019 x 0.025 inch rectangular wire. Elastomeric ring ligature was not advantageous for sliding of brackets and wires in the dry state. PMID- 15886844 TI - [The influence on expression of cyclin D1 by anti-sense oligonucleotide expression vector]. AB - PURPOSE: To construct eurokaratic expression vectors of pcDNA3.1-cyclin D1 and pcDNA 3.1-Anti-cyclin D1, and evaluate their influence on the expression of cyclin D1 in stable transfected Tca8113/CDDP cells. METHODS: The full length of cyclin D1 and its antisense fragments were acquired from Tca8113/CDDP cells by RT PCR. These cDNA fragments were inserted into pUCm-T vector and the full length sequenced. Induced by IPTG, the vector could express cyclin D1 protein in vitro. After double digested by Hind III and EcoRI at the two ends of these cDNA fragments, these cDNA fragments were inserted into pcDNA3.1 vector and transfected into Tca8113/CDDP cells with the help of Lipfectamin 2000. Stable cell lines were acquired after continually selected with G418 for 4 weeks. The expression of cyclin D1 was detected with RT-PCR and immunohistochemical staining. RESULTS: pcDNA3.1-cyclin D1, pcDNA3.1-Anti-5' and pcDNA3.1-Anti-3' vectors were constructed and named as pcDNA3.1(CO), pcDNA3.1-Anti-cyclin D1 5'(C5'), pcDNA3.1-Anti-cyclin D1 3'(C3') and pcDNA3.1-cyclin D1(CD1), respectively. The expression of cyclin D1 in Tca8113/CDDP cells transfected with C3' was significantly inhibited and the inhibition rate was 58.8%. While C5' could not inhibit the expression of cyclin D1. The protein level of cyclin D1 was strongly positive in Tca8113/CDDP-C0, Tca8113/CDDP-CD1 and Tca8113/CDDP-C5' cells. It was obviously repressed in Tca8113/CDDP-3' cells. CONCLUSION: We successfully constructed pDNA3.1-cyclin D1 vector and pcDNA3.1-Anti-cyclin D1 vectors. Detected with RT-PCR and immunohistochemical staining, the mRNA level and protein expression of cyclin D1 were significantly inhibited in C3' transfected Tca8113/CDDP cells, which may provide a good experimental tool for further studying in reversing the multidrug resistance of Tca8113/CDDP cells. PMID- 15886845 TI - [The expressions of cyclooxygenase-2 (Cox-2),VEGF in oral squamous cell carcinoma and precancerous lesions and their significances]. AB - PURPOSE: To determine expression of Cox-2 in oral squamous cell carcinoma (SCC) and oral leukoplakia and the relation to VEGF. METHODS: Immunohistochemical staining was used to quantify Cox-2 and VEGF expressions in specimens from 48 patients with oral SCC (including 20 well differentiated cases, 16 moderately differentiated cases and 12 low differentiated cases), 35 patients with oral leukoplakia (epithelial dysplasia), and 20 patients with normal oral mucosa. All specimens were analyzed by conformation quantitative assay system, their stain strength was calculated. Statistical analysis of ANOVA and linear correlation was performed using SAS statistical software. RESULTS: The expressions of Cox-2 and VEGF increased significantly as the progression of oral mucosa from normal to oral leukoplakia and oral SCC. The expression of Cox-2 reached peak in well differentiated SCC (0.167+/-0.004), and its expression decreased sharply as differentiation became poor. On the other hand, the expression of VEGF reached peak in well differentiated carcinoma (0.143+/-0.014). The expressions of Cox-2 and VEGF had negative correlation in differently differentiated SCC tissues (r=0.649, P<0.001). CONCLUSION: The expressions of Cox-2 and VEGF were significantly up-regulated in the early carcinogenesis of oral mucosa, and they may be influenced each other. Thus, they were important in the carcinogenesis and progression of oral SCC. PMID- 15886846 TI - [Curve analysis and geometric measurement of root canal curvatures]. AB - PURPOSE: To study the anatomical features of curved root canals. METHODS: According to Nagy's classification,the root canals were divided into 4 types:type I (straight), type C (continuously curved), type J (apically curved) and type S (multicurved). Each type had a sample selected from extracted permanent teeth. Digital radiographs were taken at buccolingual and mesiodistal directions with a K-file inserted into the root canal. The images were treated by Photoshop7.0 and CorelTRACE10 softwares, after that a CAD/CAM software UG NX V1.0 was applied to extract the 2D curves of the root canal axis from the images,then the 3D curves of the root canal axis were built up accordingly. The curvature of the curves was analysed and the geometric parameters were measured by UG. RESULTS: The curvature values of type I root canal were very low; the curvature of type C root canal reached a peak on the middle part,it decreased slowly towards the two ends; type J root canal had a curvature peak near the root apex, and the middle 1/3 and neck 1/3 parts of the root canal were "curvature flat zones". Type S root canal had a curvature inflection,and beside it were 2 curvature peaks at opposite directions. The geometric parameter values of the same root canal's 2D and 3D axis were different. CONCLUSION: Each point on the root canal axis had a curvature value and a vector of tangent, different root canal forms had different rules of curvature distribution, just 1 or 2 parameters couldn't describe a curved root canal completely. PMID- 15886847 TI - Cell biology in orthodontic tooth movement: the known and the unknown. AB - This review is aimed at providing, at the cellular level, a concise and complete overview of the important knowledge on the mechanism of orthodontic tooth movement to orthodontists and postgraduates who are involved or interested in basic research. The construction of this article was oriented to the following key questions: Where an osteoclast starts to its first resorption site? When the cascade of a resorption cycle starts? What are the factors involved in bone remodeling and how they orchestrate? What happens before and after the formation of a resorption pit? Major findings in these aspects were summarized and discussed. In addition, related biological phenomenon such as orthodontically induced root resorption was intensively reviewed. By means of an updated and systematic review, the author intended to introduce more biological evidence to orthodontic intervention and to encourage evidence-based treatment in daily orthodontic practice. PMID- 15886848 TI - [Timing of treatment with bite-jumping appliance in orthodontics]. AB - Bite-jumping appliance is a rational appliance for patients with severe overjet and retrusive mandible. In the pubertal growth spurt of patient, the appliance has good effect on skeletal or dental Angle class II malocclusions. After functional treatment, the probability of orthodontic treatment and orthopedic surgery in adult can be reduced. However, the most suitable time to use the bite jumping appliance has been discussed continuously in various studies. According to many studies related, this paper summarized the treatment time of the fixed and the removable bite-jumping appliances, and compared the advantages and disadvantages between them. PMID- 15886849 TI - [Nursing of root canal therapy with nickel titanium mechanical instruments]. AB - PURPOSE: To discuss the related problems of nursing during root canal preparation with nickel titanium mechanical instruments. METHODS: 206 cases were nursed who underwent root canal preparation, after which the dentists and patients were surveyed on their satisfactory extents, in order to assess the effect of nursing. RESULTS: The results of root canal preparation of 206 cases were good without any accidents such as break of instruments, while 100% dentists and 97.58% patients were satisfied with the nursing. CONCLUSION: Proper and skilled nursing could shorten treatment time, enhance patients' compliableness and decrease the unnecessary injuries. Thus it raised the efficiency and therapeutic effect. PMID- 15886850 TI - [Oral lymphoepithelial cyst: report of 3 cases]. AB - Three cases with oral lymphoepithelial cyst were reported in this paper. The most frequent site of the cyst was mouth floor, and the cyst was often misdiagnosed as mucous cyst and other diseases of cyst. Histopathologically, the central cystic lesion was lined with stratified squamous epithelium. Simultaneously, there were lymphoid cells located at the surrounding of the cystic fibrous connective tissue. Oral lymphoepithelial cyst was regarded to originate from secretory duct of the sublingual gland and the small salivary gland epithelium. With chronic stimuli, the epithelium of secretory duct was squamous metaplasia. The deciduous cells in the inner side of the duct induced the infiltration of the surrounding lymphocytes. The relevant literatures were reviewed regarding to the histological origin and the pathogenesis of the disease to provide some references for diagnosis and treatment of this disease. PMID- 15886851 TI - [Resection of carotid body tumor under interrupted block of common carotid artery]. AB - 7 patients with carotid body tumor (CBT) were evaluated by DSA preoperatively. Carotid artery (CA) blocking test was carried out under local anesthesia before operation. With CA blocked temporarily, tumors were resected under general anesthesia after the test was normal. Meanwhile some cases underwent restoration with CA repair, end-to-end anastomosis or artificial vascular grafting. No case underwent internal bypass. CA was blocked 2 to 4 times in operation. The average blocking time of CA was 22 minutes and the longest blocking time of that reached 28 minutes. The average blood loss of cases without embolization preoperatively was 1100 ml, cases with embolization was 480 ml. 3 cases had nerve complications. DSA is of important value in the diagnosis and treatments of CBT. Temporary blocking artery method can help to judge when to use internal bypass, make operation easy, decrease blood loss and nerve complications. PMID- 15886852 TI - [Use of Z plasty for ankyloglossia: report of 38 cases]. AB - Z plasty was used in 38 children (male 22, female 16) aged from 0.5 to 6 years with ankyloglossia. There were no infection, dehiscence and tongue adhesion. The wound healed more quickly, and tongue protrusion improved from 13.5 mm preoperatively to 24.6 mm postoperatively. Similarly, tongue elevation improved from 4.8 to 20.6 mm. PMID- 15886853 TI - The lethal impacts of Roundup and predatory stress on six species of North American tadpoles. AB - The decline in amphibians across the globe has sparked a search for the causes, and recent evidence suggests a connection with pesticides. However, for most pesticides, tests on amphibians are rare and conducted only for short durations (1 to 4 days) and without natural stressors. Recent studies have discovered that the stress of predator cues in the water can make insecticides much more lethal to larval amphibians, but it is unknown whether this phenomenon can be generalized to other types of pesticides. Using six species of North American amphibian larvae (Rana sylvatica, R. pipiens, R. clamitans, R. catesbeiana, Bufo americanus, and Hyla versicolor), I examined the impact of a globally common herbicide (Roundup) on the survival of tadpoles for 16 days with and without the chemical cues emitted by predatory newts (Notophthalmus viridescens). LC50(16-d) estimates varied from 0.55 to 2.52 mg of active ingredient (AI)/L, which was considerably lower than the few previous studies using Roundup (1.5 to 15.5 mg AI/L). Moreover, in one of the six species tested (R. sylvatica), the addition of predatory stress made Roundup twice as lethal. This discovery suggests that synergistic interactions between predatory stress and pesticides may indeed be a generalizable phenomenon in amphibians that occurs with a wide variety of pesticides. PMID- 15886854 TI - Toxicity of a phytosterol mixture to grayling (Thymallus thymallus) during early developmental stages. AB - The study concerns the toxicity of a phytosterol mixture, ultrasitosterol, consisting mainly of beta-sitosterol 75.7% and beta-sitostanol 13.0%, to grayling (Thymallus thymallus) embryos. Eyed eggs were exposed to three concentrations (1 microg/l, 10 microg/l, and 50 microg/l) of ultrasitosterol for 4 weeks. Embryos and later on hatched fry were taken for triiodothyronine (T3), thyroxine (T4), and histopathological analyses after 7, 14, 21, and 28 days of exposure. Most of the eggs (>95%) hatched during the first week of exposure, and ultrasitosterol treatment shortened hatching time significantly (p < or = 0.0001) at all exposure concentrations in comparison to the control. Ultrasitosterol did not have any significant effect on T3 or T4 levels in the embryo extracts. However, an interesting observation was that T3 levels increased in all treatments and in the control near the time of hatching. In conclusion, ultrasitosterol showed potential to affect the development of grayling embryos and fry, but further long term exposure experiments are needed to verify these changes in more detail. PMID- 15886855 TI - [Shock wave treatment for tennis elbow]. AB - Randomized controlled trials were evaluated to assess the effectiveness of extracorporeal shock wave treatment in the management of tennis elbow. Five trials had a mediocre methodology and four trials had a high-quality design. Well designed randomized control trials have provided evidence of the effectiveness of shock wave intervention for tennis elbow. PMID- 15886856 TI - [A simplified technique for repair of quadriceps tendon rupture by transpatellar PDS-cord]. AB - Quadriceps tendon ruptures are relatively unusual injuries caused by direct or more frequently indirect trauma. Since complete ruptures lead to loss of active extension of the knee joint, operative treatment is usually indicated. Several techniques are described in the literature. However, relatively little is known about the functional outcome after operative treatment of acute quadriceps tendon ruptures. We present a new operative technique using a 1.3-mm PDS cord passed through a transverse drill hole in the proximal pole of the patella. We operated ten consecutive cases of complete quadriceps tendon ruptures with the technique described between January 2000 and June 2003. Eight of ten patients were evaluated after a mean follow-up time of 38 months by physical examination, IKDC Subjective score, Lysholm and Tegner score as well as an isokinetic test of the quadriceps strength. No complications were noted in this period. The average postoperative scores were 87 (IKDC), 98 (Lysholm), and 4.5 (Tegner). Isokinetic testing showed an average of 25% quadriceps strength deficit. The operative treatment of complete quadriceps tendon ruptures using a PDS cord through a drill hole in the patella is a safe and effective technique permitting functional postoperative treatment. PMID- 15886857 TI - Silica structure in the spicules of the sponge Suberites domuncula. AB - Accumulation of silica in marine organisms such as diatoms and sponges has been widely reported. The proteins depositing silica in these organisms have been identified and its structure has also been described. The ultrastructure of silica has not been studied in detail, however. Herein we describe the structure of silica in the spicules of the sponge Suberites domuncula. Peroxide treatment was performed to remove the organic compounds, thereby enabling a better study of the silica. Methods used for the study included scanning and transmission electron microscopy. Electron diffraction enabled structural comparison with silica glass at the atomic level. Small-angle X-ray scattering (SAXS) of the spicules was also conducted and structure correlation between these methods attempted. At a lower magnification, spicule needles with a smooth outer surface were visible. Diffraction results suggested a network-like structure in the spicules. Silica particles of 3 nm diameter could be measured by SAXS. PMID- 15886858 TI - Determination of hexavalent chromium by using speciated isotope-dilution mass spectrometry after microwave speciated extraction of environmental and other solid materials. AB - Precise and accurate determination of hexavalent chromium in different types of solid environmental sample is regarded as a technical challenge with significant potential error if historically accepted methods are used. Microwave-assisted alkaline extraction (0.5 mol L(-1) NaOH + 0.28 mol L(-1) Na2CO3) followed by anion-exchange chromatographic separation and inductively coupled plasma mass spectrophotometric detection has been shown to provide accurate and precise results. To obtain a better understanding of potential species conversion during and/or after extraction steps, speciated isotope-dilution mass spectrometry (SIDMS) (EPA Method 6800) metrology has been successfully applied as a diagnostic tool with the modified accompanying extraction version of EPA Method 3060A. In our study, aggregate materials distributed over a large area of a major western US state were found to contain a high concentration of total chromium (195 +/- 13 to 709 +/- 19 microg g(-1)) and significant amounts of Cr6+ (141 +/- 6 to 341 +/- 29 microg g(-1)) which are at least three orders of magnitude higher than the US EPA threshold limit (0.5 microg g(-1)). Sediment samples from a major western US state, studied independently, were found to contain less (1.77 +/- 0.34 microg g( 1)) or no Cr6+ in the presence of significant total chromium. PMID- 15886860 TI - Short-term effects on bone turnover of replacing milk with cola beverages: a 10 day interventional study in young men. AB - In the Western world, increased consumption of carbonated soft drinks combined with a decreasing intake of milk may increase the risk of osteoporosis. This study was designed to reflect the trend of replacing milk with carbonated beverages in a group of young men on a low-calcium diet and studies the effects of this replacement on calcium homeostasis and bone turnover. This controlled crossover intervention study included 11 healthy men (22-29 years) who were given a low-calcium basic diet in two 10-day intervention periods with an intervening 10-day washout. During one period, they drank 2.5 l of Coca Cola per day and during the other period 2.5 l of semi-skimmed milk. Serum concentrations of calcium, phosphate, 25-hydroxycholecalciferol, 1,25-dihydroxycholecalciferol (1,25(OH)2D), osteocalcin, bone-specific alkaline phosphatase (B-ALP) and cross linked C-telopeptides (CTX), plasma intact parathyroid hormone (PTH) and urinary cross-linked N-telopeptides (NTX) were determined at baseline and endpoint of each intervention period. An increase in serum phosphate (P<0.001), 1,25(OH)2D (P<0.001), PTH (P=0.046) and osteocalcin (P<0.001) was observed in the cola period compared to the milk period. Also, bone resorption was significantly increased following the cola period, seen as increased serum CTX (P<0.001) and urinary NTX (P<0.001) compared to the milk period. No changes were observed in serum concentrations of calcium or B-ALP. This study demonstrates that over a 10 day period high intake of cola with a low-calcium diet induces increased bone turnover compared to a high intake of milk with a low-calcium diet. Thus, the trend towards a replacement of milk with cola and other soft drinks, which results in a low calcium intake, may negatively affect bone health as indicated by this short-term study. PMID- 15886861 TI - High-dose glucocorticoid treatment induces rapid loss of trabecular bone mineral density and lean body mass. AB - A recent large-scale study revealed that glucocorticoid treatment increased fracture risk, which occurred at a far smaller dose and by a shorter duration than previously thought. To study the underlying mechanism for the increased risk of fracture, we studied the early changes in bone mineral density (BMD) and body composition by dual energy X-ray absorptiometry (DXA) after initiating high-dose glucocorticoid treatment. High-dose glucocorticoid treatment was arbitrarily defined as daily doses of >or=40 mg of a predonisolone equivalent. The 33 patients enrolled in this study had not received glucocorticoid treatment before. Only 2 months of treatment resulted in substantial BMD loss, most markedly in the lumbar spine, followed by the femoral neck and total body, which suggests the preferential trabecular bone loss. Body composition was also greatly affected. Thus, 2-month treatment with glucocorticoid significantly reduced bone mineral content (BMC), lean body mass (LBM) and increased fat mass (FAT). Our results are likely to have some clinical relevance. First, BMD loss occurs quite rapidly after starting glucocorticoid treatment, and patients receiving glucocorticoid treatment should be more carefully monitored for their BMD. Second, LBM, which mainly represents muscle volume, decreases rapidly after initiating glucocorticoid treatment. Decreased LBM might be also responsible for the increased risk of fracture, since falling is a well-known risk factor for fracture, and patients receiving glucocorticoid treatment should also be evaluated for their body composition. PMID- 15886862 TI - Bone mineral density measures in longitudinal studies: the choice of phantom is crucial for quality assessment. The Tromso study, a population-based study. AB - Determination of change in bone mineral density (BMD) requires high-precision densitometry techniques. The purpose of the study is to investigate to what degree different densitometer phantoms reflect observed changes in human BMD and to investigate to what degree fluctuations in densitometers' measurement level influence bone loss estimates. Densitometer influence was assessed using the aluminum forearm phantom (AFP) provided by the manufacturer, the European forearm phantom (EFP) of semi-anthropomorphic calcium-hydroxyapatite, and repeated population measurements on different densitometer combinations. The mean follow up time was 6.4 years (SD 0.6). Measured population bone loss varied from 4.6%/year to 3.2%/year, depending on densitometer combinations. These variations could not be explained by differences in sex, age, height, weight and baseline BMD. They were predicted by EFP measurements, but not AFP measurements. The EFP measurements indicate that X-ray tube replacement changed the densitometers' measurement level in one of three instances, whereas "wear and tear" did not. We used the EFP data for adjustment of the densitometers' measurement levels. After adjustment, the overall crude bone loss was reduced from 4.14% to 3.92%. Mean annual loss was reduced from 0.64% or 0.61%. We conclude that densitometer performance might influence the accuracy of bone loss estimates. Changes in performance are not detected by aluminum phantoms. Quality control of BMD measurements in longitudinal studies should be performed with anthropomorphic calcium-hydroxyapatite phantoms in order to detect possible differences between the participating densitometers' measurement levels. PMID- 15886863 TI - The (CA)n polymorphism of the TNFR2 gene is associated with peak bone density in Chinese nuclear families. AB - Low peak bone density (PBD) in adulthood is an important determinant of osteoporotic fracture (OF) in the elderly. The tumor necrosis factor receptor 2 (TNFR2) gene has been considered as an important candidate gene for PBD due to its important role in bone turnover. In this study, we recruited a total of 1,263 subjects from 402 Chinese nuclear families composed of both parents and at least one daughter, and tested the association of the (CA)(n) polymorphism in intron 4 of the TNFR2 gene with PBD using a more contemporary quantitative transmission disequilibrium test (QTDT). Significant within-family association was detected between the CA16 allele and bone mineral density (BMD) at the lumbar spine with the P-value of 0.005 after permutations, which is still significant after correction for multiple testing. Some evidence of total-family association between the CA16 allele and lumbar spine BMD was found (P=0.021), although the significant level did not reach the empirical threshold (P< or =0.007). About 3.14% of lumbar spine BMD variation can be explained by the CA16 allele. In summary, our results suggest that the TNFR2 gene may play an important role in determining lumbar spine BMD variation in Chinese women. PMID- 15886864 TI - Food-exchange with humans in brown capuchin monkeys. AB - To assess how brown capuchin monkeys (Cebus apella) delay gratification and maximize payoff, we carried out four experiments in which six subjects could exchange food pieces with a human experimenter. The pieces differed either in quality or quantity. In qualitative exchanges, all subjects gave a piece of food to receive another of higher value. When the difference of value between the rewards to be returned and those expected was higher, subjects performed better. Only two subjects refrained from nibbling the piece of food before returning it. All subjects performed two or three qualitative exchanges in succession to obtain a given reward. In quantitative exchanges, three subjects returned a food item to obtain a bigger one, but two of them nibbled the item before returning it. Individual differences were marked. Subjects had some difficulties when the food to be returned was similar or equal in quality to that expected. PMID- 15886865 TI - Elastic properties of polymer networks with sliding junctions. AB - This paper proposes simple models of polymer networks with sliding junctions for molecular simulation and reports the main results obtained by Brownian dynamics on the elastic properties of networks with tri-functional sliding junctions. The stress-strain relation for isotropic swelling and uniaxial deformation are obtained and compared with those of the conventional chemical gels. We find that mobility and distribution of sliding junctions along the polymer chains drastically change with deformation, and lead to new profiles of the stress. We also find that sliding junctions aggregate by deformation, resulting in the decrease in the number of elastically effective chains. PMID- 15886866 TI - Parental recall of pre-school behavior related to ADHD and disruptive behavior disorder. AB - The aim of this study was to examine the contribution of Age of Onset Criterion (AOC) to the diagnosis of Attention Deficit Hyperactivity Disorder (ADHD) and disruptive behavior disorder. For this purpose, a 10-item Likert-type Parent Assessment of Pre-school Behavior Scale (PARPS), developed by the experimenters, was used to examine the presence of ADHD related pre-school behaviors in a sample of 246 children. Factor analysis and correlational analysis imply the continuity of ADHD and disruptive behavior disorder from pre-school to the elementary school years. Discriminant analysis was used to examine the diagnostic capabilities of PARPS, both by itself, and in conjunction with a battery of widely used diagnostic and clinical ADHD and disruptive behavior scales. The results of this study suggest that PARPS is a short and user-friendly scale that can contribute to the examination of the presence of AOC in ADHD and disruptive behavior disorder. PMID- 15886867 TI - Anxiety sensitivity in children of panic disorder patients. AB - Anxiety sensitivity (AS), which refers to the tendency to interpret anxiety related bodily sensations as having potentially harmful somatic, psychological or social consequences, has been proposed as a vulnerability factor for the development of panic disorder (PD). The current study examined the anxiety sensitivity levels in children of parents with panic disorder. Children of panic disorder patients (n = 68) and children of healthy parents (n = 68) filled out the Childhood Anxiety Sensitivity Index, while parents completed the Anxiety Sensitivity Index. Children of parents with panic disorder did not display higher levels of anxiety sensitivity than children of healthy parents. Furthermore, no association between anxiety sensitivity levels of parents with panic disorder and their children was found. Anxiety sensitivity is not clearly manifest in children of parents with panic disorder and might be a developing vulnerability factor that may increase towards late adolescence or early adulthood. PMID- 15886868 TI - Gender differences in delinquent behavior among Korean adolescents. AB - The present study examined gender differences in the rate, type, and relevant variables underlying delinquent behavior among South Korean adolescents. Although female delinquency is increasing and becoming more violent in South Korea, the rate of delinquent behavior was found to be much lower among female than among male adolescents and female adolescents were much less involved in antisocial, aggressive, and psychopathic delinquent behavior compared to male adolescents. Moreover, compared to female delinquent adolescents, male delinquent adolescents were found to have greater tendencies towards antisocial personality, sociability, being sexually abused, and alcohol and drug use. In contrast, female delinquent adolescents had a greater tendency toward depression than male delinquent adolescents. No gender differences were found in the association between family dynamics and delinquent behaviors. Age and antisocial personality had the most significant total effects on male delinquent behavior. In contrast, alcohol and drug abuse was the strongest contributing factors in female delinquent behavior, although the level of alcohol and drug abuse was much higher among male adolescents than among female adolescents. PMID- 15886869 TI - Behavioral and developmental characteristics of children with inversion of chromosome 9 in Korea: a preliminary study. AB - The purpose of this study is to examine the behavioral and developmental characteristics of children with inv(9). This case control study included 12 inv(9) subjects and 45 normal students. All of the subjects, together with their parents, underwent a psychiatric interview and parent questionnaire consisting of a Child Behavior Checklist (CBCL). The mean scores with regard to the social problems and total problems profiles of the CBCL were significantly higher in the inv(9) group than in the normal control group. The inv(9) group exhibited language and motor developmental delay. These findings suggest that there is a possibility of inv(9) being associated with child developmental or behavioral problems. PMID- 15886870 TI - Gender appropriateness of symptom criteria for attention-deficit/hyperactivity disorder, oppositional-defiant disorder, and conduct disorder. AB - We examined the gender appropriateness of the DSM-IV symptoms of attention deficit/hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), and conduct disorder (CD). In Study 1, 100 mothers (35 of children with and 65 of children without ADHD) rated how gender-typical and problematic they saw DSM-IV symptoms of ADHD, ODD, and CD; feminine descriptions of ADHD, ODD, and CD behaviors that we created; and relationally and overtly aggressive behaviors. Mothers rated the DSM-IV symptoms and overt aggression as boy-descriptive, and the feminine items that we created and relational aggression as girl-descriptive. Mothers saw the feminine items as less problematic than the masculine items. In Study 2, for 80 girls (40 with and 40 without ADHD), mothers' ratings on the feminine items were related to the corresponding DSM-IV symptoms, and to general psychopathology and impairment. Most correlations were significant and support the construct validity of the feminine items. PMID- 15886871 TI - How young children treat objects and people: an empirical study of the first year of life in autism. AB - OBJECTIVE: To figure out features of autism before the age of one and to explore the pathways of early social and nonsocial attention in autism through home movies. METHOD: Home movies of 15 children later diagnosed with autism, are compared with home movies of 13 typical children. The films of the two groups have been mixed and rated by blind observers through a Grid composed of social and nonsocial item and applied to two age ranges: 0-6 months (T1) and 7-12 months (T2). Two MANOVAs, an ANOVA and discriminant analyses were applied. RESULTS: Significant differences between the two groups were found only for the item in the Social area at T1 but not at T2, when groups did not differ in either social or nonsocial areas. At T2 children with autism had significantly higher scores in the nonsocial area while normal children did not show significant differences between areas. Discriminant analyses revealed that social attention can distinguish the two groups at T1 but not at T2. CONCLUSIONS: The fundamental impairment of joint attention in autism could be considered a consequence of the early atypical developmental gap and of a later disconnection between attention to people and objects. Abnormal developmental trajectories for social and nonsocial attention could help us in the future to understand relationships between adaptive capacities and symptoms, and set the stage for appropriate early screening instruments. PMID- 15886872 TI - Nitric oxide modulates hepatocyte growth factor/scatter factor-induced angiogenesis. AB - OBJECTIVE: There is limited knowledge about potential therapeutic targets in Hepatocyte growth factor/scatter factor (HGF)-induced pathophysiological angiogenesis. Recent candidates have included phosphatidylinositol-3-kinase, which is an upstream activator for endothelial nitric oxide (NO) synthase (NOS III). The current study is the first to evaluate the possible involvement of NOS NO cascade in HGF-induced angiogenesis. METHODS AND RESULTS: NOS III inhibitors blocked the HGF-induced functional neovascularization in vivo, as quantified using vessel counts, 133Xe-clearance, and immunohistology. This was reversed by L arginine. Western blot analysis of HGF-treated cells also revealed a temporal increase in HGF-induced phosphorylation. In a deconstructional approach, HGF induced the proliferation and chemokinesis of human endothelial cells. These phenotypic effects were inhibited by NOS inhibitors, L-NAME and L-NIO, and the NO scavenger, carboxy PTIO, but unaltered by 1400W, a NOS II inhibitor. This inhibition was reversed by spermine NONOate, a NO donor, which independently exerted a biphasic effect on endothelial cell proliferation. The modulation of NO did not alter HGF-induced chemoinvasion of endothelial cells, while spermine NONOate destabilized HGF-induced tubulogenesis, suggesting that a single assay is not sufficient for predicting the final phenotypic outcome on angiogenesis. CONCLUSIONS: The study is the first to demonstrate that the NOS III nitric oxide is a key signal cascade in HGF-induced angiogenesis, and represents a promising target for the clinical management of pathological conditions characterized by overt HGF signaling. PMID- 15886873 TI - Endothelial network formed with human dermal microvascular endothelial cells in autologous multicellular skin substitutes. AB - A human skin equivalent from a single skin biopsy harboring keratinocytes and melanocytes in the epidermal compartment, and fibroblasts and microvascular dermal endothelial cells in the dermal compartment was developed. The results of the study revealed that the nature of the extracellular matrix of the dermal compartments plays an important role in establishment of endothelial network in vitro. With rat-tail type I collagen matrices only lateral but not vertical expansion of endothelial networks was observed. In contrast, the presence of extracellular matrix of entirely human origin facilitated proper spatial organization of the endothelial network. Namely, when human dermal fibroblasts and microvascular endothelial cells were seeded on the bottom of an inert filter and subsequently epidermal cells were seeded on top of it, fibroblasts produced extracellular matrix throughout which numerous branched tubes were spreading three-dimensionally. Fibroblasts also facilitated the formation of basement membrane at the epidermal/matrix interface. Under all culture conditions, fully differentiated epidermis was formed with numerous melanocytes present in the basal epidermal cell layer. The results of the competitive RT-PCR revealed that both keratinocytes and fibroblasts expressed VEGF-A, -B, -C, aFGF and bFGF mRNA, whereas fibroblasts also expressed VEGF-D mRNA. At protein level, keratinocytes produced 10 times higher amounts of VEGF-A than fibroblasts did. The generation of multicellular skin equivalent from a single human skin biopsy will stimulate further developments for its application in the treatment of full-thickness skin defects. The potential development of biodegradable, biocompatible material suitable for these purposes is a great challenge for future research. PMID- 15886874 TI - Targeting neovascular pericytes in neurofibromatosis type 1. AB - Apart from tumor-driven neovascularization, a less-appreciated consequence of neurofibromatosis type 1 (NF1) is the hyperproliferation of vascular mural cells (pericytes). This study aims at establishing a role for pericytes in NF1, and determining whether interference with the function of a key pericyte component (NG2 proteoglycan) inhibits NF1 tumor neovascularization. Neovascularization in NF1 was studied in Nf+/+(control), Nf1+/-, and Nf1-/-embryos at E-10, ischemia induced retinal angiogenesis model in 24 eyes of Nf1+/-, Nf1+/+mice, and in malignant peripheral nerve sheath tumors (MPNSTs) derived from NF1 patients (ST88 14, NMS-2PC) orthotopically grown in nude mice (Crl: nu/nu). The anti-angiogenic effect of intracorneal polymer pellets containing anti-NG2 neutralizing antibody was quantified in the nude-mouse corneal angiogenesis model in which angiogenesis was induced by xenografting NMS-2PC tumor into the corneal stroma of 22 eyes. By using confocal microscopy, immunohistochemistry, and BrdU proliferation assay, the pericyte/endothelium ratios and proliferation rates were measured. Activated pericytes were present at the leading tip of the angiogenic sprouts. Pericytes showed continuous investment of endothelium in both NMS-2PC and ST88-14 MPNST tumor xenografts. Mean corneal angiogenesis induced by NMS-2PC tumor grafts in NG2-antibody treated eyes was 1.491 and 3.186 mm2 in isotype-matched non immunoglobulin treated eyes (control) (P=0.0002). A total of 193.8 vascular nuclei (a measure of ischemia-induced retinal angiogenesis) was present in angiogenic retinal tufts in Nf1+/- mice compared to 89.23 in Nf1+/+ mice (control) (P<0.0001). Mean pericyte/endothelium investment ratios were 1.015, 1.380, and 2.084 in control, Nf1+/-, and Nf1-/-embryos, respectively. Pericytes were 23% (control), 49% (Nf1+/-), and 69% (Nf1-/-) BrdU-positive. Endothelial cells from the same embryos were 29% (control), 47% (Nf1+/-), and 62% (Nf1-/-) BrdU-positive. Angiogenesis is accelerated in NF1 due to hyperproliferation of pericytes and endothelial cells. Mitotically activated NG2-positive pericytes, and endothelial cells may serve as potential therapeutic targets in NF1. PMID- 15886875 TI - VEGF treatment induces signaling pathways that regulate both actin polymerization and depolymerization. AB - The angiogenic growth factor vascular endothelial growth factor (VEGF) enhances endothelial cell migration through the activation of multiple signaling transduction pathways. Actin reorganization is an important component in VEGF induced migration, yet the signaling pathways mediating this process remain unclear. Actin reorganization involves both actin polymerization and depolymerization, and in this study we demonstrate that VEGF-treatment regulates both of these activities. With respect to actin polymerization, our results indicate that the actin nucleation promoting factors (NPF) neural Wiskott-Aldrich syndrome protein (N-WASP) binds the SH2- plus SH3-domain containing adaptor protein Nck in both control and VEGF-treated cells. We had previously showed that VEGF treatment leads to the recruitment of Nck to activated receptor, and our current results indicate a VEGF-dependent redistribution of N-WASP to the cell surface. A Nck dominant-negative blocked Nck recruitment to receptor, blocked N WASP cellular redistribution and attenuated actin stress fiber formation. With respect to actin depolymerization, VEGF-treatment led to the rapid phosphorylation of the actin depolymerization factor cofilin, and its upstream regulator, LIM-kinase (LIMK). Unlike what is observed in certain other cell types, the p21-activated kinase (PAK), a Nck binding protein, does not mediate VEGF-induced LIMK phosphorylation, as a PAK dominant-negative had no effect on this activity. The PAK dominant-negative also did not affect VEGF-induced actin reorganization. Pharmacological inhibitors of phosphoinositide-3 kinase (PI3-K) and the rho-activated kinase (ROCK) attenuated VEGF-induced LIMK phosphorylation, indicating a role for (PI3-K) and ROCK in the signaling pathways leading to regulation of LIMK activity. PMID- 15886876 TI - Anti-migratory and anti-angiogenic effect of p16: a novel localization at membrane ruffles and lamellipodia in endothelial cells. AB - Recent evidence has established different functions for the tumor suppressor protein, p16(INK4A) aside from controlling the cell cycle. Here we report that cdk4/6 inhibition blocked both human umbilical vein endothelial cells (HUVEC) spreading on a vitronectin matrix and HUVEC migration on vitronectin. p16 can also act as an anti-angiogenic molecule in vitro since HUVEC and HMEC cells transfected with Ad-p16 or treated with Antennapedia p16 peptides are unable to differentiate on a Matrigel matrix. Both, p16, cyclin D1, cdk4 and cdk6 were immuno-colocalized with Ezrin, Rac, Vinculin, alphav-integrin, and FAK proteins in the ruffles and lamellipodia of migratory cells. Our results indicate that p16 is a key component of a new cytoplasmic pathway controlling angiogenesis of endothelial cells via the alphavbeta3-integrin-mediated migration. PMID- 15886877 TI - Biological activity of bevacizumab, a humanized anti-VEGF antibody in vitro. AB - Bevacizumab (Avastin, Genentech) is a humanized monoclonal antibody targeting vascular endothelial growth factor (VEGF), a critical angiogenic factor involved in both physiological and pathological conditions. It has been recently approved by the US FDA as a first-line therapy for widespread metastatic colorectal cancer. This report is a detailed biological characterization of bevacizumab in a variety of in vitro models. It is shown that bevacizumab potently neutralizes VEGF and blocks its signal transduction through both the VEGFR-1 and VEGFR-2 receptors, as demonstrated by the inhibition of VEGF-induced cell proliferation, survival, permeability, nitric oxide production, as well as migration and tissue factor production. Although bevacizumab retains the ability to bind to human Fcgamma receptors and complement protein C1q, it does not demonstrate cell or complement-mediated cytotoxicity in either VEGF producing or targeting cells. Thus the mechanism of anti-tumor activity of bevacizumab is most likely due to its anti-angiogenesis effect through binding and neutralization of secreted VEGF. PMID- 15886878 TI - The VEGF receptor tyrosine kinase inhibitor, ZD6474, inhibits angiogenesis and affects microvascular architecture within an orthotopically implanted renal cell carcinoma. AB - The proangiogenic vascular endothelial growth factor-A (VEGF) is essential for the development of new tumor vessels. ZD6474 is a novel inhibitor of VEGF receptor-2 (VEGFR-2) tyrosine kinase activity, which also has additional activity against epidermal growth factor (EGF) receptor tyrosine kinase. The antitumor activity of different schedules of ZD6474 in a clinically relevant, metastasizing, murine renal cell carcinoma (RENCA) model was evaluated in this study. RENCA cells were inoculated into the left kidney of 24 mice (day 0). Daily ZD6474 (50 mg/kg p.o.) treatment was initiated 1 day or 10 days after tumor cell inoculation and continued until day 21. Following treatment, kidney weight and volume were assessed and blood vessel density determined by CD31 staining. Visible metastases in the lungs, spleen, and lymph nodes were quantified using a dissection microscope. In an additional study, animals were treated according to the same regimen and quantitative three-dimensional microvascular corrosion casting was performed to enable detailed assessment of the tumor vascular architecture. Therapy initiated on day 1 or day 10 resulted in a 79% and 59% reduction in primary tumor volume, a 79% and 60% reduction in the number of lung metastases, and a 58% and 59% reduction in vessel density of primary tumors compared with the control group, respectively. Corrosion casting proved a 5.4- and 3.2-fold lower vascular volume compared with untreated tumors, observations that paralleled with significant architectural alterations. In this RENCA model, ZD6474 was a highly active inhibitor of tumor angiogenesis, primary tumor growth and tumor metastasis. PMID- 15886881 TI - Stem cells and tissue homeostasis in mammary glands. PMID- 15886882 TI - Bovine mammary progenitor cells: current concepts and future directions. AB - Although cell number is positively correlated with milk production, much remains to be learned about the bovine mammary stem cell and progenitor cells. Bovine mammary development is driven by many of the same classic mammogenic hormones studied in murine models, yet histologic features of bovine mammary development differ from that of rodent models. Most notably, terminal end buds, as they have been described for murine models, do not exist in the bovine mammary gland. However, among the most important common features of mammary development in disparate species is the involvement of histologically distinct, lightly staining epithelial cells as putative stem and progenitor cells. Although stem cell research has often focused on mammary development, mammary stem cells seemingly provide the basis for mammary growth and cell turnover in the mature animal. These cells provide an obvious focus for research aimed at increasing the efficiency of milk production. This review addresses recent findings concerning the histology and molecular physiology of putative bovine mammary stem and progenitor cell populations, areas where more study is critically needed, and areas where studies of bovine mammary physiology may present a unique opportunity to better understand mammary physiology in many species. PMID- 15886883 TI - Stem/progenitor cells in mouse mammary gland development and breast cancer. AB - Breast cancer is a genetically and clinically heterogeneous disease. It is unclear whether different target cells contribute to this heterogeneity and which cell types are most susceptible to oncogenesis. Stem cells are speculated to be the cellular origin of at least a subset of human breast cancers. To begin to address these issues, we have isolated and characterized cell populations enriched in normal mammary stem/progenitors and have studied the expression of putative stem/progenitor markers in tumors derived from genetically engineered mice. Specifically, transgenic activation of Wnt signaling in the mammary gland induces tumors comprised of epithelial and myoepithelial cells harboring the same genetic defect implying that the tumor arose from transformation of a bipotent progenitor cell. On the other hand, transgenic activation of Neu signaling induces tumors comprising cells of more limited lineage capacity. Thus, the heterogeneity of different breast cancers may reflect the activation of different oncogenic pathways, different cellular targets in which these genetic changes occur, or both. PMID- 15886884 TI - Pregnancy and stem cell behavior. AB - The identification of cancer-initiating epithelial subtypes (i.e. cancer stem cells) is important for gaining a more comprehensive understanding of the process of neoplastic transformation and tumorigenesis. Since reproductive history has a major impact on breast tumorigenesis, it is reasonable to assume that pregnancy and lactation have enduring effects on the cancer susceptibility of multipotent progenitors. Using the Cre-lox technology as a tool to genetically label pregnancy-hormone-responsive cells, we identified a mammary epithelial subtype that is abundant in parous females. These pregnancy-induced mammary epithelial cells (PI-MECs) originate from differentiating cells during the first pregnancy and lactation cycle. They do not undergo apoptosis during postlactational remodeling, and they persist throughout the remainder of a female's life. In this review, we discuss the biological relevance of PI-MECs in multiparous females and their important stem cell-like features, such as self renewal, as well as their ability to produce progeny with diverse cellular fates. Using appropriate animal models, we further demonstrate that PI-MECs are cellular targets for pregnancy enhanced mammary tumorigenesis. PMID- 15886885 TI - The mammary gland "side population": a putative stem/progenitor cell marker? AB - Hematopoietic Stem Cells have been isolated by their ability to pump out Hoechst 33342 dye and form a distinct population definable by flow cytometry--the Side Population (SP). The membrane pump Bcrp has been identified as the molecular determinant of the SP phenotype. An SP population with Bcrp activity has been defined in a number of tissues, including mouse mammary and human breast epithelium, and it has been proposed that the SP phenotype is a universal stem cell marker. Studies of mouse and human breast SP suggest that the population is undifferentiated but capable of differentiating into epithelial structures of both luminal and myoepithelial lineages both in vitro and in vivo. However, evidence that the SP is enriched for stem cells is, at the moment, only correlative, and there are potentially confounding technical issues. We still await formal proof that the SP contains a stem cell population. PMID- 15886886 TI - Epithelial progenitors in the normal human mammary gland. AB - The human mammary gland is organized developmentally as a hierarchy of progenitor cells that become progressively restricted in their proliferative abilities and lineage options. Three types of human mammary epithelial cell progenitors are now identified. The first is thought to be a luminal-restricted progenitor; in vitro under conditions that support both luminal and myoepithelial cell differentiation, this cell produces clones of differentiating daughter cells that are exclusively positive for markers characteristic of luminal cells produced in vivo (i.e., keratins 8/18 and 19, epithelial cell adhesion molecule [EpCAM] and MUC1). The second type is a bipotent progenitor. It is identified by its ability to produce "mixed" colonies in single cell assays. These colonies contain a central core of cells expressing luminal markers surrounded by cells with a morphology and markers (e.g., keratin 14(+)) characteristic of myoepithelial cells. Serial passage in vitro of an enriched population of bipotent progenitors promotes the expansion of a third type of progenitor that is thought to be myoepithelial-restricted because it only produces cells with myoepithelial features. Luminal-restricted and bipotent progenitors can prospectively be isolated as distinct subpopulations from freshly dissociated suspensions of normal human mammary cells. Both are distinguished from many other cell types in mammary tissue by their expression of EpCAM and CD49f (alpha6 integrin). They are distinguished from each other by their differential expression of MUC1, which is expressed at much higher levels on the luminal progenitors. To relate the role of these progenitors to the generation of the three-dimensional tubuloalveolar structure of the mammary tree produced in vivo, we propose a model in which the commitment to the luminal versus the myoepithelial lineage may play a determining role in the generation of alveoli and ducts. PMID- 15886887 TI - Maintenance of cell type diversification in the human breast. AB - Recent genome-wide expression analysis of breast cancer has brought new life to the classical idea of tumors as caricatures of the process of tissue renewal as envisioned by Pierce and Speers (Cancer Res 1988;48:1996-2004) more than a decade ago. The search for a cancer founder cell or different cancer founder cells is only possible if a hierarchy of differentiation has been established for the particular tissue in question. In the human breast, the luminal epithelial and myoepithelial lineages have been characterized extensively in situ by increasingly elaborate panel of markers, and methods to isolate, culture, and clone different subpopulations have improved dramatically. Comparisons have been made with the mouse mammary gland in physiological three-dimensional culture assays of morphogenesis, and the plasticity of breast epithelial cells has been challenged by immortalization and transformation. As a result of these efforts, several candidate progenitor cells have been proposed independently of each other, and some of their features have been compared. This research has all been done to better understand breast tissue homeostasis, cell-type diversification in general and breast cancer evolution in particular. The present review discusses the current approaches to address these issues and the measures taken to unravel and maintain cell type diversification for further investigation. PMID- 15886888 TI - Survival of mammary stem cells in suspension culture: implications for stem cell biology and neoplasia. AB - There is increasing evidence that a variety of neoplasms including breast cancer may result from transformation of normal stem and progenitor cells. In the past, isolation and characterization of mammary stem cells has been limited by the lack of suitable culture systems able to maintain these cells in an undifferentiated state in vitro. We have recently described a culture system in which human mammary stem and progenitor cells are able to survive in suspension and produce spherical colonies composed of both stem and progenitor cells. Recent observation that adult stem cells from other tissues may also retain the capacity for growth under anchorage independent conditions suggests a common underlying mechanism. We propose that this mechanism involves the interaction between the canonical Wnt signal pathway and E-cadherin. The Wnt pathway has been implicated in normal stem cell self-renewal in vivo. Furthermore, there is evidence that deregulation of this pathway in the mammary gland and other organs may play a key role in carcinogenesis. Thus, the development of in vitro suspension culture systems not only provides an important new tool for the study of mammary cell biology, but also may have important implications for understanding key molecular pathways in both normal and neoplastic stem cells. PMID- 15886889 TI - Dietary factors modifying breast cancer risk and relation to time of intake. AB - Multiple factors contribute to the development of human breast cancer. However, environmental factors, especially dietary factors, appear to have the greatest effects. Evidence obtained in epidemiological studies has been corroborated by laboratory findings. Dietary components strongly associated with breast cancer include fat and phytochemicals. A diet high in n-3 polyunsaturated fatty acid (PUFA) or monounsaturated fatty acid (MUFA) and low in n-6 PUFA is protective against breast cancer. Some phytochemicals present in fruits and vegetables are also protective. Time of intake appears to be important: lifetime protection may be achieved if one is exposed to a dietary factor that lowers breast cancer risk early in life. Synergistic and antisynergistic interactions between dietary factors can modify breast cancer risk. The available evidence suggests that breast cancer risk can be reduced by early dietary intervention. PMID- 15886890 TI - Measures of resting immune function and related physiology in juvenile rainbow trout exposed to a pulp mill effluent. AB - Juvenile rainbow trout (Oncorhynchus mykiss) were exposed to 0, 10, 30, and 70% (v/v) of a secondary-treated, integrated thermomechanical-bleached kraft pulp and paper mill effluent. The immunological parameters oxidative burst and phagocytosis of head kidney macrophages and total and differential circulating leukocytes were measured after 21 d of exposure. General parameters of stress and exposure including erythrocyte counts, numbers of degenerating erythrocytes, splenic pigmented macrophage aggregates (PMAs), spleen size, bile chemistry, and hepatic EROD activity were also assessed. Contrary to parallel chronic studies on the same effluent, EROD induction did not occur. Analyses of bile indicated uptake and accumulation of resin acids and some sterols. There was no measurable macrophage-related immunological dysfunction. However, circulating leukocytes, specifically lymphocytes, were reduced. The density of splenic PMAs increased over the exposure period, possibly in association with degenerating blood cells. There were statistical differences between staggered days of sampling in head kidney oxidative burst, white and mature red blood cell counts, and spleen size, indicating that relatively minor capture and handling stress could result in rapid changes in some parameters. Overall, it was concluded that the observed minor, indirect alterations in the immune response were likely the result predominantly of a nonspecific mechanism such as a cortisol-mediated stress response. PMID- 15886891 TI - Residues of organic chlorinated pesticides in agricultural soils of Beijing, China. AB - Concentration of organic chlorinated pesticides (OCPs) were measured in topsoils of a selected farm (NK farm) in Beijing in 1993 and 2003. The results indicated that OCPs, mainly 1,2,3,4,5,6-hexachlorocyclohexane (HCH) and 1,1,1,-trichloro 2,2-bis(p-chlorophenyl) ethane (DDT), degraded greatly in the 20 years after the prohibition of their use. DDT was the major contributors of pollution on the farm with 92.23% and 81.28% contributions of total OCP load in soils in 2003 and 1993 respectively. The levels of total DDT and HCH in the old orchard group, in which cultivation began in 1962, were significantly higher (p < 0.05) than those in the new orchard group in which cultivation began in 1991, and all data were higher than the level of barren land (p < 0.05). Studying the isomeric and parent substance metabolite ratios indicated there was more application and accumulation of DDT in the old orchard and that DDT in the new orchard had undergone a different degradation time period or perhaps had been applied more recently, but no new input of HCH was detected. Pollution potential was assessed on the basis of China Soil Environmental Quality Standard. PMID- 15886892 TI - Urinary 8-hydroxydeoxyguanosine in Belarussian children relates to urban living rather than radiation dose after the chernobyl accident: a pilot study. AB - As a result of the Chernobyl accident in 1986, exposure to radioactive cesium is still a concern in the contaminated regions of Belarus. We tested the hypothesis that long-term radiation exposure from the Chernobyl accident might increase the urinary excretion of the oxidative stress marker, 8-hydroxydeoxyguanosine (8 OHdG), in Belarussian children. Urinary 8-OHdG was determined in two groups of children (-n = 31 and n = 46) -living in contaminated and uncontaminated areas of Belarus, respectively (the majority of the unexposed children lived in the capital Minsk). The children from the contaminated areas had a significantly higher annual summary effective dose but significantly lower urinary 8-OHdG levels than the children from the uncontaminated areas. Unexpectedly, children living in uncontaminated urban areas had significantly higher urinary 8-OHdG levels than children living in uncontaminated rural areas. There was no statistically significant effect of sex or body mass index on urinary 8-OHdG, but there was a weak significant inverse correlation to age as well as to the annual summary effective dose. These findings suggest that radiation from the Chernobyl accident is now a less important contributor to oxidative stress in Belarussian children than urban living. PMID- 15886893 TI - Survey of human exposure to PCDDs, PCDFs, and coplanar PCBs using hair as an indicator. AB - Our environment is polluted with toxic compounds including polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (Co-PCBs). We investigated the suitability of hair analysis for testing human exposure to toxic compounds. Hair analysis revealed that municipal solid-waste (MSW) incineration workers were exposed to PCDDs, PCDFs, and Co-PCBs at high levels. The total 2,3,7,8-TCDD toxic equivalent concentration in MSW incineration workers was 2.5 times higher than in the general population, thus indicating that the indoor environment of the incineration facility was more polluted than the general environment. There were also characteristic patterns of occurrence of these dioxin-like compounds in the hair of smokers from the general population. We confirmed that hair analysis is useful for evaluating human exposure to PCDDs, PCDFs, and Co-PCBs in the atmosphere. PMID- 15886894 TI - Antioxidant enzymes and tissue regeneration in Eurythoe complanata (Polychaeta: Amphinomidae) exposed to used vehicle crankcase oil. AB - Polychaetes, Eurythoe complanata, from the Gulf of Cariaco,Venezuela, were exposed to 0.3, 1.6, and 3.3% water-soluble fraction (WSF) of used crankcase oil during 15 and 21 days. The antioxidant enzymes glutathione peroxidase (GPX), glutathione reductase (GR), catalase (CAT), and glutathione-S-transferase (GST) were assayed in the body wall tissue. Furthermore, after chemical exposure, the polychaetes were cut into equal halves; then wound healing and the number of regenerated body segments were recorded periodically. GST activity was affected by all the experimental treatments, with activity increasing with WSF concentrations. GPx activity was altered for the contamination period. GR and CAT activities rose in response to increasing WSF concentrations, and were higher for long-term than for short-term exposures. The wound healing of the transected body regions was retarded by WSF exposure. WSF affected the tissue regeneration, which was almost abolished at 3.3% WSF. The exposure period did not affect the tissue repairing responses. Alteration of GST in contaminated organisms suggested equivalent changes in detoxication of bioaccumulated organic contaminants. The variation of GR and CAT suggests induction of oxidative stress that could reduce the ability of WSF-exposed worms to repair damaged tissue. PMID- 15886895 TI - Differences in DDT and PCB residues between common and striped dolphins from the southwestern Mediterranean. AB - Organochlorine concentrations (OCs) and stable isotopes were investigated in the blubber of common dolphins (Delphinus delphis) and striped dolphins (Stenella coeruleoalba) from the southwestern Mediterranean. Samples were obtained from dolphins entangled in fishing nets during the 1992-1994 fishing season and from biopsies taken in 1992. Intrapopulation variations were studied, but because most of the dolphins were juveniles or calves (90%), no significant differences were found on the basis of reproductive condition or sex. Only mature male common dolphins showed significantly higher levels of most of the compounds studied than immature individuals did. There were quantitative and qualitative interspecific differences in organochlorine compounds profile. As compared to common dolphins, striped dolphins carried higher concentrations of organochlorine concentrations (OCs), their %DDE/tDDT and PCB/tDDT ratios were significantly higher, and recalcitrant PCB congeners were more abundant. Distribution and information on composition of stomach contents would in principle support a higher exposure to OCs in common dolphins as compared to striped dolphins, thus apparently contradicting the observed results. However, stable isotopes showed that striped dolphins exploit a higher trophic level, thus explaining observed differences. Interspecific dissimilarities in metabolic capacity to handle OCs may be an added factor. Although in the two species OC concentrations exceeded levels considered to be ineffective in marine mammals, pollution-induced effects on populations could not be properly assessed. PMID- 15886896 TI - Toxic effects of bisphenol A on sexual and asexual reproduction in Hydra oligactis. AB - Hydra oligactis, an evolutionarily primitive invertebrate, produced eggs or testes (sexual reproduction) when starved at 10 degrees C, and produced buds (asexual reproduction) when fed at 20 degrees C. Bisphenol A (BPA) at 2-4 mg/L given to male or female hydra had adverse effects on both sexual and asexual reproduction. Despite the estrogenic nature of BPA, testis formation and egg formation were similarly affected. The doses causing these acute toxicities were comparable to those reported earlier in aquatic invertebrates and were much higher than environmentally detected doses, at which the disruption of the endocrine system has been reported in fishes. All these facts indicate that the adverse effects are the results of general toxicity and may not be due to the estrogenic function of the compound. On the other hand, we found that BPA at 1 mg/L (a dose still much higher than environmental doses) stimulated asexual reproduction. No such stimulation of sexual reproduction was seen. When male hydras were fed at 10 degrees C, they produced both buds and testes simultaneously. BPA at 0.5 and 1 mg/L under this condition also stimulated asexual reproduction, whereas it suppressed sexual reproduction more severely than BPA at 2-3 mg/L. There may be some interaction between processes involved in sexual and asexual reproduction under this condition, and the stimulation of asexual reproduction by BPA may cause suppression of sexual reproduction. PMID- 15886897 TI - Using transgenic Caenorhabditis elegans in soil toxicity testing. AB - Soil bioassays are important tools for evaluating toxicological effects within the terrestrial environment. The American Society for Testing and Materials E2172 01 Standard Guide outlines a method for conducting laboratory soil toxicity tests using the nematode Caenorhabditis elegans. This method is an efficient tool for extracting C. elegans from soil samples and can be carried out after a 24-h exposure period using relatively small amounts of soil. Drawbacks of this method include problems with (1) recovery of nematodes from soils containing a high percentage of organic matter, and (2) distinguishing indigenous nematode species from nematodes added for the laboratory test. Due in part to these issues, C. elegans has not been extensively accepted for use in soil testing. To address these concerns and improve upon the American Society for Testing and Materials method, this project focused on using transgenic strains of C. elegans carrying a GFP-expressing element. Lethality and behavior tests revealed that the transgenic nematodes respond similarly to the wild-type N2 strain, indicating that they can be used in the same manner in soil testing. The GFP marker is easily identifiable not only within soils containing a large amount of organic matter, but also in field-collected soils containing indigenous nematodes. These results support the use of transgenic GFP C. elegans in soil bioassays as a tool to further the reliability of laboratory toxicity tests. PMID- 15886898 TI - Analysis of UNEP priority POPs using HRGC-HRMS and their contamination profiles in livers and eggs of great cormorants (Phalacrocorax carbo) from Japan. AB - The present investigation demonstrates establishment of United Nations Environment Programme (UNEP) priority Persistent Organic Pollutants (POPs) using high-resolution gas chromatography-high-resolution mass spectrometry. Particularly, POPs analytical methods were established using native and (13)C labeled internal standards of HCHs, HCB, cyclodienes, chlordanes, DDTs, mirex, dioxin-like PCBs, PCDDs, and PCDFs by isotope dilution technique. The relative response factor for 6-point calibration curve native standards (18 replicate analysis) were in the ranges of 0.93-1.43 with relative standard deviation ranges from 1.68 to 4.96%. Instrument detection limit and instrument quantification limit was established for various POPs at femtograms. Concentrations of UNEP-POPs were measured in liver (n = 10) and egg (n = 10) of great cormorants and their major diet, gizzard shad (n = 2), collected in and around Tokyo, Japan. DDTs (ranges in liver and egg, respectively) were predominant accumulants (9800 310,000 and 9600-73,000) followed by dioxin-like PCBs (4500-69,000 and 7900 150,000), chlordanes (2600-16,000 and 700-4,800), cyclodienes (650-4600 and <1 1000), HCB (680-2800 and 180-590), HCHs (230-1800 and 120-490), PCDD/DFs (3.2-27 and 1.7-5.7) on nanogram per gram lipid basis. Concentrations (ranges) of POPs in gizzard shad were in the following order: DDTs (3900-16,000), chlordanes (3400 14,000), cyclodienes (340-1300), HCB (110-480), and HCHs (140-360) on nanogram per gram lipid basis. PMID- 15886899 TI - Metal kinetics and respiration rates in F1 generation of carabid beetles (Pterostichus oblongopunctatus F.) originating from metal-contaminated and reference areas. AB - We investigated resistance to metals in carabid beetles inhabiting metal-polluted and reference areas. Chronic multigeneration exposure to toxic metal concentrations may potentially result in adaptation through decreased metal uptake rate and/or increased excretion rate. The cost of resistance to pollution could be associated with increased metabolic rate. To test these predictions, laboratory cultured F(1)-generation beetles originating from metal-polluted and reference sites were exposed to food contaminated with zinc and/or cadmium for 10 weeks. After that, uncontaminated food was offered to the animals for another 3 weeks. During the experiment, internal concentrations of Cd and Zn were measured as were respiration rates of the animals. The results obtained show no significant differences in metal accumulation and excretion patterns or respiration rates between the populations. This may suggest that adaptation has not occurred in the beetles chronically exposed to toxic metal concentrations. The possible explanations for the lack of differences between the populations are discussed. PMID- 15886900 TI - Estrogenic and dioxin-like activities and cytotoxicity of sediments and biota from Hong Kong mudflats. AB - Persistent organic pollutants, such as organochlorine insecticides and polychlorinated biphenyls (PCBs), were measured in several environmental matrices including aerial deposition, seawater, sediment, and biota in two important coastal wetlands of Hong Kong, China. Specifically, samples were collected from within the Mai Po Marshes Nature Reserve (Mai Po), an internationally acclaimed wetland situated in the northwestern part of the New Territories of Hong Kong, and A Chau in Starling Inlet, a relatively remote island on the eastern side of Hong Kong. Hexachlorobenzene, dichlorodiphenyltrichloroethanes, and hexachlorocyclohexanes were detected in all samples collected from Mai Po. Environmental endocrine disruptors (including dioxin-like compounds and estrogenic chemicals), measured by the use of cell-based chemical activated luciferase expression assays, were found to occur at concentrations that might pose a risk to the ecologic systems in Mai Po. Dioxin-like PCBs were detected at small concentrations in some of the samples. Concentrations of 2,3,7,8 tetrachlorodibenzo-p-dioxin equivalents (TEQs) were primarily related to the relatively great concentrations (>100 ng/g dry weight) of high molecular-weight polycyclic aromatic hydrocarbons in sediments, whereas the relative proportion of TEQs contributed by nonortho-substituted PCBs was small. Polar compounds primarily contributed estrogen equivalents, which were measured in sediments. Significant concentrations of cytotoxic compounds were detected in fish samples collected from the Mai Po but not in fish collected from A Chau. PMID- 15886901 TI - A motion for cautious optimism in the treatment of patients with ocular melanoma: the ayes have it. PMID- 15886902 TI - Selecting the highly selective. PMID- 15886903 TI - The role of liver resections for noncolorectal, nonneuroendocrine metastases: experience with 142 observed cases. AB - BACKGROUND: To evaluate the role of liver resection for noncolorectal, nonneuroendocrine metastases, indications and results were retrospectively reviewed in 142 observed patients. METHODS: A curative liver resection was performed in 83 cases (58.5%), and the remaining 59 patients received palliative treatments. The primary tumor site was gastrointestinal in 18, breast in 21, genitourinary in 15, leiomyosarcoma in 10, and other in 19. The mean number of metastases was 1.4. The mean diameter of the nodules was 5.7 cm. Liver metastases were synchronous in 11 (13.3%) cases and metachronous in the remaining 72 (86.7%). RESULTS: There was no operative mortality. Postoperative morbidity was 20.5%. The median postoperative stay was 9.5 days. The 3- and 5-year actuarial survival rate was 49.5% and 34.3% in resected cases, respectively, whereas there were almost no survivors 3 years after diagnosis in unresected cases (P < .05). The 3- and 5-year disease-free survival was 41.4% and 23.8%, respectively. Among the 83 resected cases, the 3- and 5-year actuarial survival was 17.3% and 8.6% for metastases from gastrointestinal tumors, 53.9% and 24.6% from breast cancer, 63.7% and 36.4% from leiomyosarcoma, 50.4% and 37.8% from genitourinary neoplasms, and 55.6% and 42.4% from other sites, respectively. Fifteen patients (18.1%) survived longer than 5 years. CONCLUSIONS: Liver resection is an effective treatment for noncolorectal, nonneuroendocrine metastases; it allows satisfactory long-term survival with an acceptable operative risk in selected patients. Hepatic metastases from gastrointestinal carcinoma have the worst prognosis; those from genitourinary tumors show a better outcome. Patient selection is the key to achieving encouraging results. PMID- 15886904 TI - Treatment of liver metastases from uveal melanoma. AB - BACKGROUND: Uveal melanoma patients with liver metastases have a poor prognosis. The effect of screening and multimodality treatment (including surgery) should be evaluated. METHODS: A total of 602 patients treated for uveal melanoma during a 14-year period had abdominal ultrasonography screening every 6 months. Sixty three developed liver metastases as the first extraocular metastatic site. When possible, liver surgery and intra-arterial catheter implantation were performed. The influence on survival of demographics, uveal tumor characteristics, liver metastasis presentation, and treatment was studied. RESULTS: The median time to liver metastasis was 29 months. Twenty-eight patients (44%) were operated on: 14 (22%) had R0 liver surgery, and 14 with diffuse liver involvement had R2 liver surgery (there were no significant surgical complications). Thirty-five patients with diffuse liver involvement received systemic chemotherapy or best supportive care only. The median overall survival was 15 months (range, 3-110 months): 25 months for the 14 patients with R0 surgery, 16 months for the 14 with R2 surgery, and 11 months for the 35 with chemotherapy or supportive care. By univariate analysis, age (< or =70 years), number of metastases (< or =10), and quality of operation (R0) were predictive of a better prognosis. CONCLUSIONS: In the case of liver metastases from uveal melanoma, aggressive treatment permitting tumor eradication seems to offer a chance of long-term survival to selected patients. Nevertheless, neither ultrasound screening nor quality of operation had an effect on the outcome of most patients (78%). Better screening tests and more effective multimodality treatments are required to improve survival in uveal melanoma patients with hepatic metastases. PMID- 15886905 TI - Outcome in 846 cutaneous melanoma patients from a single center after a negative sentinel node biopsy. AB - BACKGROUND: A negative sentinel node biopsy (SNB) implies a good prognosis for melanoma patients. The purpose of this study was to determine the long-term outcome for melanoma patients with a negative SNB. METHODS: Survival and prognostic factors were analyzed for 836 SNB-negative patients. All patients with a node field recurrence were reviewed, and sentinel node (SN) tissue was reexamined. RESULTS: The median tumor thickness was 1.7 mm, and 23.8% were ulcerated. The median follow-up was 42.1 months. Melanoma specific survival at 5 years was 90%, compared with 56% for SN-positive patients (P < .001). On multivariate analysis, only thickness and ulceration retained significance for disease-free and disease-specific survival. Five-year survival for patients with nonulcerated lesions was 94% vs. 78% with ulceration. Eighty-three patients (9.9%) had a recurrence. Twenty-seven patients developed recurrence in the regional node field, and in 22 of these, it was the first recurrence site. Six developed local recurrence, 17 an in-transit metastasis, and 58 distant disease. The false-negative rate was 13.2%. SN slides and tissue blocks were further examined in 18 patients with recurrence in the node field, and metastatic disease was found in 3 of them. CONCLUSIONS: This large, single-center study confirms that patients with a negative SNB have a significantly better prognosis than those with positive SNs. In those with a negative SNB, primary tumor thickness and ulceration are independent predictors of survival. Incorrect pathologic diagnosis contributed to only a minority of the false-negative results in this study. PMID- 15886906 TI - Sentinel nodes are identifiable in formalin-fixed specimens after surgeon performed ex vivo sentinel lymph node mapping in colorectal cancer. AB - BACKGROUND: In recent years, the technique of sentinel lymph node (SLN) mapping has been applied to colorectal cancer. One aim was to ultrastage patients who were deemed node negative by routine pathologic processing but who went on to develop systemic disease. Such a group may benefit from adjuvant chemotherapy. METHODS: With fully informed consent and ethical approval, 37 patients with primary colorectal cancer and 3 patients with large adenomas were prospectively mapped. Isosulfan blue dye (1 to 2 mL) was injected around tumors within 5 to 10 minutes of resection. After gentle massage to recreate in vivo lymph flow, specimens were placed directly into formalin. During routine pathologic analysis, all nodes were bivalved, and blue-staining nodes were noted. These later underwent multilevel step sectioning with hematoxylin and eosin and cytokeratin staining. RESULTS: SLNs were found in 39 of 40 patients (98% sensitivity), with an average of 4.1 SLNs per patient (range, 1-8). In 14 of 16 (88% specificity) patients with nodal metastases on routine reporting, SLN status was in accordance. Focused examination of SLNs identified occult tumor deposits in 6 (29%) of 21 node-negative patients. No metastatic cells were found in SLNs draining the three adenomas. CONCLUSIONS: The ability to identify SLNs after formalin fixation increases the ease and applicability of SLN mapping in colorectal cancer. Furthermore, the sensitivity and specificity of this simple ex vivo method for establishing regional lymph node status were directly comparable to those in previously published reports. PMID- 15886907 TI - Quality of life and functional long-term outcome after partial pancreatoduodenectomy: pancreatogastrostomy versus pancreatojejunostomy. AB - BACKGROUND: To determine the effects of pancreatogastrostomy (PG) versus pancreatojejunostomy (PJ) as types of reconstruction after partial pancreatoduodenectomy on postoperative quality of life and long-term gastrointestinal morbidity, the outcomes of 104 patients (PG, n = 63; PJ, n = 41) were evaluated. METHODS: To compare the two groups, the European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire (QLQ-PAN 26) standard and an additional self-developed questionnaire were used. The mean time after surgery was 6.4 +/- 3.4 years. RESULTS: In the PG group, there was a significant reduction of gastric acid reflux, gastroduodenal ulcers, and pain compared with before surgery. However, a significant increase in steatorrhea, intolerance toward larger meals, and aversion against certain foods were observed. In the PJ group, no significant change of preoperative symptoms was present except for jaundice. The incidence of diabetes mellitus and the need for pancreatic enzyme substitution had increased significantly but similarly in both groups. The global quality of life was identical in both groups of patients. CONCLUSIONS: This analysis demonstrates that the global quality of life was not affected by the type of reconstruction after partial pancreatoduodenectomy. Patients who underwent PG had a significant reduction of gastric reflux, pain, and abdominal discomfort compared with before surgery. Patients in both groups showed an impaired exocrine and endocrine pancreatic function of a similar extent. PMID- 15886908 TI - Constitutive expression of small heat shock protein in an htpG disruptant of the Cyanobacterium Synechococcus sp. PCC 7942. AB - In cyanobacteria, a disruptant of hspA encoding a small heat shock protein homologue, shows decreased cell growth rates at moderately high temperatures, and loss of both basal and acquired thermo-tolerances, which resemble the phenotype of an htpG disruptant. In vitro studies have shown that both small heat shock protein and Hsp90 can bind and keep non-native proteins in a refolding-competent state under denaturing conditions. The aim of the present study is to elucidate whether constitutive expression of HspA can functionally replace HtpG, a prokaryotic homolog of Hsp90, in the cyanobacterium Synechococcus sp. PCC 7942. HspA did not improve the viability of the htpG disruptant at a lethal temperature, although it did that of the wild type. It did not improve an iron starved phenotype of the mutant under normal growth conditions, a novel phenotype found in the present study. These results suggest that cellular function of HtpG may differ significantly from that of HspA. PMID- 15886909 TI - Virulence of Pseudomonas aeruginosa cells surviving photodynamic treatment with toluidine blue. AB - We report results of a study carried out to investigate photodynamic effect of Touildine blue (TBO) on virulent factors secreted by Pseudomonas aeruginosa. Viability of MCF-7 cells incubated with culture supernatants of bacteria photosensitized with TBO (experimental) was found to be higher than that for MCF 7 cells incubated with culture supernatants of bacterial cells treated either with TBO in dark (control II) or not receiving any treatment (neither TBO nor light (control I)). Furthermore, whereas MCF-7 cells incubated with supernatants of bacteria, control I and II, showed significant morphological alterations, no such changes were observed in MCF-7 cells incubated with supernatant of experimental cells. The activities of phospholipase C and proteases, virulent factors of P. aeruginosa, were also found to decrease on photosensitization. These results suggest that the potency of virulent factors is reduced in cells surviving photodynamic treatment with TBO and this may have implication in treatment of infections caused by P. aeruginosa. PMID- 15886910 TI - Characterization of a novel Cr6+ reducing Pseudomonas sp. with plant growth promoting potential. AB - The isolate RNP4 obtained from a long-term tannery waste contaminated soil was characterized and presumptively identified as Pseudomonas sp. The strain RNP4 tolerated concentrations up to 450 mg Cr(6+)/L on a Luria-Bartani (LB) agar medium and reduced a substantial amount of Cr(6+) to Cr(3+) in the LB liquid medium. The ability of performing multifarious activities in tandem suggested the uniqueness of isolate RNP4. The strain produced a substantial amount of indole acetic acid (IAA) in tryptophan-supplemented medium. The strain also exhibited the production of siderophore and solubilization of phosphorus in mineral salt medium and SRS1 medium, respectively. Concurrent production of IAA and siderophore and the solubilization of phosphorus revealed its plant growth promotion potential. Furthermore, the strain was able to promote the growth of black gram, Indian mustard, and pearl millet in the presence of Cr(6+). Thus, the innate capability of this novel isolate for parallel bioremediation and plant growth promotion has significance in the management of environmental and agricultural problems. PMID- 15886911 TI - Identification, cloning, and expression of Pseudomonas aeruginosa phosphorylcholine phosphatase gene. AB - Pseudomonas aeruginosa phosphorylcholine phosphatase (PChP) is a periplasmic enzyme produced simultaneously with the hemolytic phospholipase C (PLc-H) when the bacteria are grown in the presence of choline, betaine, dimethylglycine or carnitine. Molecular analysis of the P. aeruginosa mutant JUF8-00, after Tn5-751 mutagenesis, revealed that the PA5292 gene in the P. aeruginosa PAO1 genome was responsible for the synthesis of PChP. The enzyme expressed in E. coli, rPChP-Ec, purified by a chitin-binding column (IMPACT-CN system, New England BioLabs) was homogeneous after SDS-PAGE analysis. PChP was also expressed in P. aeruginosa PAO1-LAC, rPChP-Pa. Both recombinant enzymes exhibited a molecular mass of approximately 40 kDa, as expected for the size of the PA5292 gene, and catalyzed the hydrolysis of phosphorylcholine, phosphorylethanolamine, and p nitrophenylphosphate. The saturation curve of rPChP-Ec and rPChP-Pa by phosphorylcholine revealed that these recombinant enzymes, like the purified native PChP, also contained the high- and low-affinity sites for phosphorylcholine and that the enzyme activity was inhibited by high substrate concentration. PMID- 15886912 TI - Boophilus microplus infection by Beauveria amorpha and Beauveria bassiana: SEM analysis and regulation of subtilisin-like proteases and chitinases. AB - Beauveria bassiana is a well-known broad-range arthropod pathogen which has been used in biological control of several pest insects and ticks such as Boophilus microplus. Beauveria amorpha has both endophytic and entomopathogenic characteristics, but its capacity for biological control has still not been studied. During the processes of host infection, B. bassiana and B. amorpha produce several hydrolytic extracellular enzymes, including proteases and chitinases, which probably degrade the host cuticle and are suggested to be pathogenicity determinants. To access the role of these enzymes during infection in the tick B. microplus, we analyzed their secretion during fungus growth in single and combined carbon sources, compared to complex substrates such as chitin and B. microplus cuticle. Chitin and tick cuticle-induced chitinase in both fungus and protease was induced only by tick cuticle. SEM analysis of B. amorpha and B. bassiana infecting B. microplus showed apressorium formation during penetration on cattle tick cuticle. PMID- 15886913 TI - Isolation and functional characterization of siderophore-producing lead- and cadmium-resistant Pseudomonas putida KNP9. AB - Heavy metals, being phytotoxic, cause growth inhibition and even plant death. Siderophore-producing bacterial strain KNP9 is growth promoting and has been isolated from Panki Power Plant, Kanpur, India. It simulated significant (p > 5%) root and shoot growth of mung bean to the extent of 16.48% and 28.80%, respectively in the presence of CdCl(2) (110 microM: ). However, the increase in root and shoot growth was 20% and 19.5%, respectively, in the presence of (CH(3)COO)(2)Pb (660 microM: ). Moreover, concentration of accumulated lead and cadmium in root and shoot was also reduced in the presence of this isolate ranging from 37.5 to 93.19%. A moderate reduction in chlorophyll content (39.14%) in the presence of 110 microM: CdCl(2) was rescued by bioinoculant KNP9. However, the 19.58% decrease in chlorophyll content in the case of lead acetate remained unchanged even in the presence of KNP9. Nevertheless, 16S ribosomal DNA (rDNA) sequencing identified KNP9 as a strain of Pseudomonas putida. PMID- 15886916 TI - Genetic variation in clones of Pseudomonas pseudoalcaligenes after ten months of selection in different thermal environments in the laboratory. AB - The random amplification of polymorphic DNA (RAPD) method was used to examine genetic variation in experimental clones of Pseudomonas pseudoalcaligenes in two experimental groups, as well as their common ancestor. Six clones derived from a single colony of P. pseudoalcaligenes were cultured in two different thermal regimes for 10 months. Three clones in the Control group were cultured at constant temperature of 35 degrees C and another three clones in the High Temperature (HT) group were propagated at incremental temperature ranging from 41 to 47 degrees C for 10 months. A total of 45 RAPD primers generated 146 polymorphic markers. Analysis of molecular variance (AMOVA) revealed mild (11%) but significant (P < 0.001) genetic difference between the Control and the HT clones. Phylogenetic analysis based on pairwise genetic distances showed that the HT clones were more divergent from the ancestor and from each other than the Control clones, implying that the HT clones of P. pseudoalcaligenes may have evolved faster than the Control clones. PMID- 15886918 TI - A ferric dicitrate uptake system is required for the full virulence of Bacillus cereus. AB - Bacillus cereus is an opportunistic human pathogen of increasing prevalence. Analysis of the Bacillus cereus genome sequence identified a potential ferric dicitrate uptake system. The three-gene operon was confirmed to be negatively regulated by the ferric uptake repressor (Fur). The Fec operon was genetically silenced using the integration suicide vector pMUTIN4. The mutant strain displayed no growth defect under iron-limited conditions but was unable to grow on ferric citrate as a sole iron source. The virulence of the mutant strain was attenuated in a lepidopteran infection model, highlighting the importance of iron uptake systems to the virulence of B. cereus and the potential of these systems to act as targets for novel antimicrobial agents. PMID- 15886921 TI - Coniferyl alcohol, a lignin precursor, stimulates Rhizobium rhizogenes A4 virulence. AB - Rhizobium rhizogenes, a soil bacterium, is the causative agent of the neoplastic disease hairy root. Upon incubation of Rhizobium rhizogenes A4 with coniferyl alcohol, a lignin precursor, bacterial virulence on cotton cotyledon slices was stimulated. This was observed both in numbers of root hairs produced and in their length. Stimulation was maximized after exposure of bacteria to 150 microg/mL of coniferyl alcohol for 4 h. This was shown to be at the early log phase of bacterial growth. PMID- 15886926 TI - Successful endovascular treatment of a left common carotid artery aneurysm following failed surgery of a right common carotid artery aneurysm. AB - Aneurysm of the common carotid artery is a rare and serious disease requiring prompt treatment in order to avoid neurologic complications. A 39-year-old man presented with voice impairment and a pulsatile mass at the right side of his neck and was found by color Doppler examination to have bilateral common carotid artery aneurysms of unknown origin. The right-sided large aneurysm was treated with placement of an 8 mm interposition Gore-Tex graft between the right common and internal carotid arteries. The surgical graft thrombosed 7 days after the surgery but the left-sided aneurysm was successfully treated by a Jostent peripheral stent-graft. Color Doppler examination showed a patent stent and no filling of the aneurysm on his first and sixth-month follow-up. Bilateral common carotid artery aneurysm is an exceptionally unusual condition and endovascular treatment of carotid artery aneurysms with covered stents may become an effective treatment alternative for these lesions. PMID- 15886928 TI - Palliation of painful perineal metastasis treated with radiofrequency thermal ablation. AB - We report a case of painful perineal metastasis from urinary bladder carcinoma in a 73-years-old woman, treated with CT-guided radiofrequency ablation (RFA). The pain was immediately relieved and follow-up at 1 and 6 months showed total necrosis of the mass. One year later, the patient has no pain and her quality of life is improved. PMID- 15886929 TI - Rupture of the renal artery after cutting balloon angioplasty in a young woman with fibromuscular dysplasia. AB - A 24-year-old woman with uncontrollable high blood pressure for 3 months had significant stenosis of the left renal artery caused by fibromuscular dysplasia (FMD). The lesion was resistant to percutaneous transluminal angioplasty at 18 atm with a semicompliant balloon. Angioplasy with a 6 x 10 mm cutting balloon (CB) caused rupture of the artery. Low-pressure balloon inflation decreased but did not stop the leak. An attempt to place a stent-graft (Jostent; Jomed, Rangendingen, Germany) failed, and a bare, 6-mm balloon-expandable stent (Express SD; Boston Scientific, MN) was deployed to seal the leak, which had decreased considerably after long-duration balloon inflation. The bleeding continued, and the patient underwent emergent surgical revascularization of the renal artery with successful placement of a 6-mm polytetrafluoroethylene bypass graft. CBs should be used very carefully in the treatment of renal artery stenosis, particularly in patients with FMD. PMID- 15886930 TI - Interventional radiology in liver transplantation. AB - Radiology is a key specialty within a liver transplant program. Interventional techniques not only contribute to graft and recipient survival but also allow appropriate patient selection and ensure that recipients with severe liver decompensation, hepatocellular carcinoma or portal hypertension are transplanted with the best chance of prolonged survival. Equally inappropriate selection for these techniques may adversely affect survival. Liver transplantation is a dynamic field of innovative surgical techniques with a requirement for interventional radiology to parallel these developments. This paper reviews the current practice within a major European center for adult and pediatric transplantation. PMID- 15886931 TI - Quality improvement guidelines for placement of esophageal stents. AB - Esophageal cancer is now the sixth leading cause of death from cancer worldwide. During the past three decades, important changes have occurred in the epidemiologic patterns associated with this disease. Due to the distensible characteristics of the esophagus, patients may not recognize any symptoms until 50% of the luminal diameter is compromised, explaining why cancer of the esophagus is generally associated with late presentation and poor prognosis. Esophageal cancer has a poor outcome, with an overall 5 year survival rate of less than 10%, and fewer than 50% of patients are suitable for resection at presentation. As a result palliation is the best option in this group of patients. The aims of palliation are maintenance of oral intake, minimizing hospital stay, relief of pain, elimination of reflux and regurgitation, and prevention of aspiration. For palliative care, current treatment options include thermal ablation, photodynamic therapy, radiotherapy, chemotherapy, chemical injection therapy, argon beam or bipolar electrocoagulation therapy, enteral feeding (nasogastric tube/percutaneous endoscopic gastrostomy), and intubation (self-expanding metal stents (SEMS) or semi-rigid prosthetic tubes) with different success and complications rates. PMID- 15886932 TI - Interrupted aortic arch associated with absence of left common carotid artery: imaging with MDCT. AB - Interrupted aortic arch (IAA) is a rare severe congenital heart defect defined as complete luminal and anatomic discontinuity between ascending and descending aorta. Although its association with various congenital heart defects has been reported, absence of left common carotid artery (CCA) in patients with IAA has not been reported previously. We report a case of IAA associated with the absence of left CCA which was clearly shown on multidetector-row spiral CT. PMID- 15886933 TI - Lymphatic leak complicating central venous catheter insertion. AB - Many of the risks associated with central venous access are well recognized. We report a case of inadvertent lymphatic disruption during the insertion of a tunneled central venous catheter in a patient with raised left and right atrial pressures and severe pulmonary hypertension, which led to significant hemodynamic instability. To our knowledge, this rare complication is previously unreported. PMID- 15886934 TI - Memokath metallic stent in the treatment of transplant kidney ureter stenosis or occlusion. AB - PURPOSE: To determine the efficacy of the Memokath 051 stent (Engineers and Doctors, Hornbaek, Denmark) in the treatment of recurrent ureteral stenosis or occlusion in transplant kidneys. METHODS: From October 1985 through January 2004, 1,131 renal transplantations were performed at our center. Four patients who developed recurrent renal transplant ureter obstruction had nephrostomy catheters placed. Antegrade pyelography showed ureteral stenosis in three cases and complete occlusion in one patient. In each case, a Memokath 051 stent was inserted via an antegrade approach. Mean follow-up was 20 months (range 18-21 months). Creatinine levels were measured and ultrasonography was performed during follow-up. RESULTS: All stent procedures were technically successful. During follow-up, one stent migrated within 10 days after stent insertion and was removed cystoscopically. Another stent had to be removed in the 14th month due to resistant infection, and was replaced with a new Memokath 051 stent which remained patent for another 8 months. The other two stents were fully patent at the 18th and 21st month of follow-up, respectively. CONCLUSION: Placement of a Memokath 051 stent appears to be a promising treatment alternative to balloon dilation, double-J stents and open surgical intervention for ureteral stenosis or occlusion in kidney transplant recipients. Further study of larger series is necessary. PMID- 15886935 TI - MRSA-infected external iliac artery pseudoaneurysm treated with endovascular stenting. AB - A 48-year-old woman with severe juvenile-onset rheumatoid arthritis presented with a bleeding cutaneous sinus distal to her right total hip replacement scar. Methicillin resistant Staphylococcus aureus (MRSA) was isolated on culture. She had previously undergone bilateral total hip and knee replacements at aged 23 and six years later had the right knee prosthesis removed for infection, with subsequent osteomyelitis of the femoral shaft and right total hip prosthesis disruption. Peripheral arteriography was performed in view of persistent bleeding from the sinus, which revealed a 6 cm false aneurysm filling from and compressing the right external iliac artery (EIA). A PTFE-covered, balloon expandable JOSTENT was deployed in the right EIA, successfully excluding the false aneurysm and preventing further bleeding from the sinus. No graft infection was reported at 12 months. This case illustrates the potential use of endovascular stent-grafting in the treatment of an infected pseudoaneurysm. PMID- 15886937 TI - An unusual complication of a colonic wallstent. PMID- 15886938 TI - Chronic mesenteric ischemia treated by isolated angioplasty of the inferior mesenteric artery. PMID- 15886939 TI - Percutaneous stent placement as treatment of renal vein obstruction due to inferior vena caval thrombosis. AB - A patient who had undergone his third orthotopic liver transplantation nearly 9 years prior to presentation developed worsening hepatic and renal function, as well as severe bilateral lower extremity edema. Magnetic resonance imaging demonstrated vena caval thrombosis from the suprahepatic venous anastomosis to the infrarenal inferior vena cava, obstructing the renal veins. This was treated by percutaneous placement of metallic stents from the renal veins to the right atrium. At 16 months clinical follow-up, the patient continues to do well. PMID- 15886940 TI - Detachable balloon embolization of an arterioportal fistula following liver biopsy in a liver transplant recipient: a case report and review of literature. AB - We report a case of an intrahepatic arterioportal fistula in a 61-year-old female liver transplant recipient. The patient presented with massive ascites 7 months after a percutaneous liver biopsy. A large fistula between the right hepatic artery and the right portal vein was diagnosed on color Doppler ultrasound and confirmed on arteriography. The fistula was successfully embolized with the detachable balloon technique and the ascites resolved. Symptomatic intrahepatic arterioportal fistula in a liver transplant recipient following percutaneous biopsy is rare. Clinical manifestations, surgical or endovascular therapy, and outcome are discussed. The literature on this subject is reviewed. PMID- 15886941 TI - Endovascular treatment of proximal bilateral iliac limb dislocation and kinking following endovascular abdominal aortic aneurysm repair. AB - We report the case of a 69-year-old man with a late type 1b endoleak due to proximal migration of both iliac limbs 5 years after endovascular repair of an abdominal aortic aneurysm. The endovascular method used to correct bilaterally this condition is described. Final angiographic control shows patency of the stent-graft without signs of endoleak. PMID- 15886942 TI - Radiofrequency ablated lesion in the normal porcine lung: long-term follow-up with MRI and pathology. AB - PURPOSE: To evaluate the effectiveness of magnetic resonance imaging (MRI) in reflecting histopathologic changes after radiofrequency ablation (RFA) to the lung and to assess accurately the extent of tissue necrosis for evaluating untreated lesions. METHODS: Percutaneous RFA was performed on 72 lung regions in 12 pigs under computed tomographic guidance. After performing MRI, the animals were divided into three experimental phases: in the acute phase, 4 pigs were killed immediately after the procedure; in the subacute phase, 4 pigs were killed at 1 week; and in the chronic phases, 2 pigs were killed at 4 and 8 weeks after the procedure, respectively. MRI-histopathologic correlation was performed. RESULTS: In the acute phase, the inner zone showed hypointensity on T2-weighted images and isointensity on T1-weighted images, with a lack of enhancement on contrast-enhanced T1-weighted images, representing early changes in coagulative necrosis on histopathologic examination. The outer zone showed hyperintensity on T2-weighted images and isointensity on T1-weighted images, with ring-like enhancement on contrast-enhanced T1-weighted images. The histopathologic section showed alveolar fluid collections and congestion. In the subacute phase the MR zone pattern was essentially similar to that of the acute phase, but the ablated lesion showed extensive coagulative necrosis with a fibrovascular rim on histopathologic examination. In the chronic phase, there was no change in the zone pattern on MRI. The lesions showed gradual resorption of coagulative necrosis. The area of coagulative necrosis correlated closely with the histopathologic size (p < 0.05). CONCLUSIONS: MRI effectively visualized the histopathologic changes after RFA and accurately determined the extent of the necrotic lesion. MRI is potentially a useful modality for evaluating therapeutic efficacy after RFA to the lung. PMID- 15886943 TI - Arterial embolization of unresectable hepatocellular carcinoma with use of microspheres, lipiodol, and cyanoacrylate. AB - We performed a retrospective analysis of 46 patients with histologically confirmed hepatocellular carcinoma (HCC) who were treated with transarterial embolization (TAE) of the hepatic arteries. To induce permanent embolization, microspheres (Embosphere; 100 to 700 micron) and a mixture of ethiodized oil (Lipiodol Ultrafluide) with cyanoacrylate (Glubran) was injected. A total of 106 TAE procedures were performed. Cumulative survival rates were calculated. No patient died during embolization or within the first 24 hours. Severe procedure related complications were observed in 2 patients. At the time of analysis, 38 of 46 patients were alive. The 180-, 360-, 520-, and 700-day cumulative survival rates for the total study population were 80.6%, 70.7%, 70.7%, and 47.1%, respectively, with a median survival of 666 days. TAE with the use of microspheres and Lipiodol and cyanoacrylate for unresectable HCC is a feasible treatment modality. Bland embolization with the use of microspheres can be used in patients for whom chemoembolization is not desired. PMID- 15886944 TI - MELD score as a predictor of early death in patients undergoing elective transjugular intrahepatic portosystemic shunt (TIPS) procedures. AB - PURPOSE: To Evaluate the MELD score as a predictor of 30-day mortality in patients undergoing elective TIPS procedures. METHODS: This was a retrospective, IRB-approved study. The medical records of all patients who underwent a TIPS procedure between May 1, 1999 and June 1, 2003 in a single institution were reviewed. Patients who underwent elective TIPS were selected. Elective TIPS was performed in 119 patients with a mean age of 55.1 (+/- 9.6) years. The MELD and Child-Pugh scores before TIPS, etiology of cirrhosis, portosystemic gradients before and after TIPS, procedure time, and procedural complications were obtained from the medical records. The MELD and Child-Pugh scores before TIPS were compared between the survivor group (SG) and the early death (EDG) group. The early death rate was calculated for MELD score subgroups (1-10, 11-17, 18-24, and >24). Data were analyzed using the Fisher exact test, chi-square test and independent-sample t-test. A p value of less than 0.05 was considered significant. RESULTS: Technical success rate was 100%. The early death rate was 10.9% (13/119). The mean MELD scores before TIPS were 19.4 (+/- 5.9) (EDG) and 14 (+/- 4.2) (SG) (p = 0.025). The early death rate was highest in the pre-TIPS MELD > 24 subgroup. The Child-Pugh scores were 9.0 (+/- 1.6) (SG) and 9.8 +/- 1.06 (EDG) (p = 0.08). The mean portosystemic gradients before TIPS were 20.5 (+/- 7.7) mmHg (EDG) and 22.7 (+/- 7.3) (SG) (p > 1) and the mean portosystemic gradients after TIPS were 6.5 (+/- 3.5) (EDG) and 6.9 (+/- 2.4) (SG) (p > 1). The mean procedural times were 95.6 (+/- 8.4) min (EDG) and 89.2 (+/- 7.5) min (SG) (p > 1). No early death was attributed to a fatal complication during TIPS. CONCLUSION: The MELD score is useful in identifying patients at a higher risk of early death after an elective TIPS. On th basis of our results, we do not endorse elective TIPS in patients with MELD scores > 24. PMID- 15886945 TI - Arteriomegaly. PMID- 15886946 TI - Puncture-site metastasis in a radiologically inserted gastrostomy tube: case report and literature review. AB - Gastrostomy-site metastases from head and neck cancer have been reported numerous times following endoscopic insertion, with direct implantation being implicated. We present the first reported case of gastrostomy-site metastasis following radiological insertion, and discuss the mechanisms by which this may have occurred. These include: direct implantation, hematogenous dissemination, or the natural shedding of tumor cells into the gastrointestinal tract. PMID- 15886947 TI - Balloon dilation of the superior vena cava (SVC) resulting in SVC rupture and pericardial tamponade: a case report and brief review. AB - Stent placement is an accepted primary treatment for SVC syndrome. Balloon dilation is frequently performed prior to stent placement. Although various stent related hemorrhagic complications have been reported, as well as reports of iatrogenic catheter and guidewire perforations, there has been only one previous report of balloon dilation-related SVC rupture. We report a second case, including the clinical scenario, in the hope that should this complication occur, it might be recognized quickly and treated successfully. PMID- 15886948 TI - Predicting infected bile among patients undergoing percutaneous cholecystostomy. AB - PURPOSE: Patients may not achieve a clinical benefit after percutaneous cholecystostomy due to the inherent difficulty in identifying patients who truly have infected gallbladders. We attempted to identify imaging and biochemical parameters which would help to predict which patients have infected gallbladders. METHODS: A retrospective review was performed of 52 patients undergoing percutaneous cholecystostomy for clinical suspicion of acute cholecystitis in whom bile culture results were available. Multiple imaging and biochemical variables were examined alone and in combination as predictors of infected bile, using logistic regression. RESULTS: Of the 52 patients, 25 (48%) had infected bile. Organisms cultured included Enterococcus, Enterobacter, Klebsiella, Pseudomonas, E. coli, Citrobacter and Candida. No biochemical parameters were significantly predictive of infected bile; white blood cell count >15,000 was weakly associated with greater odds of infected bile (odds ratio 2.0, p = NS). The presence of gallstones, sludge, gallbladder wall thickening and pericholecystic fluid by ultrasound or CT were not predictive of infected bile, alone or in combination, although a trend was observed among patients with CT findings of acute cholecystitis toward a higher 30-day mortality. Radionuclide scans were performed in 31% of patients; all were positive and 66% of these patients had infected bile. Since no patient who underwent a radionuclide scan had a negative study, this variable could not be entered into the regression model due to collinearity. CONCLUSION: No single CT or ultrasound imaging variable was predictive of infected bile, and only a weak association of white blood cell count with infected bile was seen. No other biochemical parameters had any association with infected bile. The ability of radionuclide scanning to predict infected bile was higher than that of ultrasound or CT. This study illustrates the continued challenge to identify bacterial cholecystitis among patients referred for percutaneous cholecystostomy. PMID- 15886949 TI - Local administration of NF-kappa B decoy oligonucleotides to prevent restenosis after balloon angioplasty: an experimental study in New Zealand white rabbits. AB - PURPOSE: To evaluate the efficacy of NF-kappa B oligonucleotides (ODN) administered by local administration with the channeled balloon catheter to prevent restenosis after balloon angioplasty in restenotic iliac arteries of New Zealand white rabbits. MATERIALS AND METHODS: In vitro, 8000 rabbit vascular smooth muscle cells (rVSMC) where transfected with a liposomal carrier (TfX50) with 100 ng of decoy and scrambled ODN. Inhibition of proliferation was measured using a MTT assay after 24 hours in comparison to control. In vivo, 22 male New Zealand White rabbits were fed a 1% cholesterol diet and received denudation of both common iliac arteries with a 3 mm balloon catheter to induce an arterial stenosis. Four weeks after stenosis induction, local application of NF-kappa B in two different concentrations (1 mug: n = 14; 10 mug: n = 8) was performed randomly on one common iliac artery. Scrambled oligonucleotides without specific binding capacities were injected into the contralateral side. The channeled balloon catheter allows simultaneous balloon dilation (8 atm) of the stenosis and local application of a drug solution (2 atm). Four weeks after local drug delivery the animals were killed and the vessels were excised and computerized morphometric measurements were performed. RESULTS: NF-kappa B decoy ODN but not scrambled ODN inhibited proliferation of rVSMC in vitro. Following local ODN application in the animals, no acute vascular complications were seen. NF-kappa B ODN resulted in a statistically non significant reduction of neointimal area compared to the control group. The neointimal area was 0.97 mm(2) using 1 mug NF kappa B ODN compared to 0.98 mm(2) in the control group. The higher dose resulted in a neointimal area of 0.97 mm(2) compared to 1.07 mm(2) at the control side. CONCLUSIONS: Local drug delivery of NF-kappa B ODN using the "channeled balloon" catheter could not reduce neointimal hyperplasia in stenostic rabbit iliac arteries. Application modalities have to be improved to enhance the effect of the local application to prevent restenosis after balloon angioplasty. PMID- 15886950 TI - Calculating colon transit time with radionuclide-filled capsules in constipated patients: a new method for colon transit study. AB - BACKGROUND: Colon motility disorders require reliable methods for calculating segmental colonic transit time. This study evaluated bowel transit time by means of a safe, easy, cheap, non-digestive, and non-disintegrating radionuclide-filled capsule that provided accurate and clear images. METHODS: Radionuclide-filled mini-containers (MCs) were prepared from infusion sets by an apparatus used for sealing blood bags or plasmapheresis sets. In vitro stability studies were performed by immersing 5% methylene blue dye-filled MCs in buffers of variable pH and enzymes simulating the conditions in the stomach and the small bowel. Colon transit scintigraphy was performed with MCs filled with iodine 131 (n = 5) and thallium 201 (n = 8) that were placed in a commercially available capsule. RESULTS: By in vitro acid, base, and intestinal enzyme resistance tests, no methylene blue leakage was determined visually and by spectrophotometric analysis. Accurate and clear images were obtained for colon transit study in constipated patients. After excretion of MCs in the feces, abdominal, myocardial, thyroid, and urinary bladder region counts were found to show the same activity as the background. Radionuclide leakage from MCs was not determined in vivo by gamma camera. CONCLUSIONS: This is a suitable, safe, easy, and cheap method to provide accurate and clear images for colon transit study in constipated patients. PMID- 15886951 TI - Endometrial cancer: magnetic resonance imaging. AB - Carcinoma of the endometrium is the most common invasive gynecologic malignancy of the female genital tract. Clinically, patients with endometrial carcinoma present with abnormal uterine bleeding. The role of magnetic resonance imaging (MRI) in endometrial carcinoma is disease staging and treatment planning. MRI has been shown to be the most valuable imaging mod-ality in this task, compared with endovaginal ultrasound and computed tomography, because of its intrinsic contrast resolution and multiplanar capability. MRI protocol includes axial T1-weighted images; axial, sagittal, and coronal T2-weighted images; and dynamic gadolinium enhanced T1-weighted imaging. MR examination is usually performed in the supine position with a phased array multicoil using a four-coil configuration. Endometrial carcinoma is isointense with the normal endometrium and myometrium on noncontrast T1-weighted images and has a variable appearance on T2-weighted images demonstrating heterogeneous signal intensity. The appearance of noninvasive endometrial carcinoma on MRI is characterized by a normal or thickened endometrium, with an intact junctional zone and a sharp tumor myometrium interface. Invasive endometrial carcinoma is characterized disruption or irregularity of the junctional zone by intermediate signal intensity mass on T2-weighted images. Invasion of the cervical stroma is diagnosed when the low signal intensity cervical stroma is disrupted by the higher signal intensity endometrial carcinoma. MRI in endometrial carcinoma performs better than other imaging modalities in disease staging and treatment planning. Further, the accuracy and the cost of MRI are equivalent to those of surgical staging. PMID- 15886952 TI - Ulcerative colitis: value of MR imaging. AB - Recent technologic advances have greatly improved the quality of abdominal magnetic resonance imaging (MRI) by allowing the identification of abnormalities in inflammatory bowel disease. Thus far, the role of MRI has been extensively investigated in Crohn disease (CD) and, to a minor extent, in ulcerative colitis (UC), likely due to intrinsic differences between these two diseases. In UC the inflammatory lesions, unlike CD, are confined to the colon, have a predictable spreading, and affect only the inner wall layer; thus endoscopy alone can assess the extent and severity of disease in most cases. However, preliminary studies have demonstrated that MRI also can be a reliable diagnostic tool for UC because it is useful for integrating clinical and endoscopic data. MRI can be valuable in distinguishing CD from UC in uncertain cases by assessing the sparing of the distal ileum and the continuity of colonic involvement. Moreover, MRI can provide important information if endoscopy is incomplete, e.g., due to tight strictures, or contraindicated, e.g., in severely acute disease, due to a high risk of perforation. MRI can detect most of the typical findings of the diseases, such as wall thickening, mural stratification, loss of haustrations, and several complications including fibrotic or neoplastic strictures. In addition, MRI can be extremely valuable in assessing disease activity by monitoring the degree of wall gadolinium enhancement and T2 signal at the level of the affected bowel segments, thus influencing pharmacologic and surgical planning. In the next few years, MRI will likely become the imaging modality of choice in the clinical management of this disease. PMID- 15886953 TI - A single-use site selection technique, using GIS, for aquaculture planning: choosing locations for mangrove oyster raft culture in Margarita Island, Venezuela. AB - Oyster culture has a potential to generate income for coastal communities and to lessen pressure on natural overexploited populations. A project to transfer mangrove oyster Crassostrea rhizophorae raft culture technology to selected coastal communities in Margarita Island, Venezuela is being developed, and an optimum location selection technique was devised. To pick the variables or factors that determine site suitability, a bibliographic database was made, aspects of interest chosen, and the most comprehensive ones singled out, eliminating redundant ones. Twenty variables were grouped in criteria based on the way they influence the project. Variables were classified as intrinsic environmental, environmental extrinsic, logistic, and socioeconomic criteria. Thirty-five experts were asked to evaluate the factors and to score each according to their suitability weight. Logistic criterion received the highest values, followed by environmental extrinsic issues. A Geographic Information System using a base map compiled from 1:25,000 scale maps was developed. A thematic map for each factor was completed, dividing graphically the 3896-km2 study area into polygons of equal weight for each factor. The Multi-Criteria Evaluation (MCE) was used to combine the variables. Resultant vectors in thematic maps were added to obtain smaller polygons with the same value sum. Finally, MCE was used to generate a final output: the optimum sites for oyster aquaculture resulting from the added values of over 3000 polygons in the maps, for the 20 criteria. Higher scores were reached in 13 areas covering 4.1 km2, those places having the optimum conditions for oyster raft aquaculture in the region. Additional locations meeting 75% to 70% of the demanded criteria for a final suitable selection cover 137 sites encompassing 37.5 km2. PMID- 15886954 TI - Toward sustainable management of national parks in Japan: securing local community and stakeholder participation. AB - Japan's national park system constitutes a potentially viable mechanism for securing local community participation and building stakeholder consensus for sustainable park management, although the potential of this system is yet to be fully maximized. This article gives an overview of the system of protecting natural resources in Japan, focusing on the national park system. Parks are managed by zoning and regulation, which is unique in that land is not "set aside" for nature conservation, but designated as national park wherever the need to preserve "scenic beauty" has been recognized, regardless of land ownership or land use. Although resource conservation under this system has been problematic, it has advantages, especially in terms of community participation. This article demonstrates that in order to reach the system's potential, the park authority must act as coordinator of stakeholders and facilitator of bottom-up approaches to decision-making. In order to do this, steps that must be taken include the following: identifying the various stakeholders in park management and defining the "local community"; clarifying the roles and responsibilities of each stakeholder; and supporting consensus-building among stakeholders regarding the objectives and long-term vision of each park. By taking these steps, it would be possible to build a park management system that overrides government boundaries and involves local communities. This will enable the park authority to address the challenges facing Japan's complex system of conserving natural resources, and move towards sustainable management of natural resources in Japan. PMID- 15886955 TI - Science, policy, and stakeholders: developing a consensus science plan for Amchitka Island, Aleutians, Alaska. AB - With the ending of the Cold War, the US Department of Energy is responsible for the remediation of radioactive waste and disposal of land no longer needed for nuclear material production or related national security missions. The task of characterizing the hazards and risks from radionuclides is necessary for assuring the protection of health of humans and the environment. This is a particularly daunting task for those sites that had underground testing of nuclear weapons, where the radioactive contamination is currently inaccessible. Herein we report on the development of a Science Plan to characterize the physical and biological marine environment around Amchitka Island in the Aleutian chain of Alaska, where three underground nuclear tests were conducted (1965-1971). Information on the ecology, geology, and current radionuclide levels in biota, water, and sediment is necessary for evaluating possible current contamination and to serve as a baseline for developing a plan to ensure human and ecosystem health in perpetuity. Other information required includes identifying the location of the salt water/fresh water interface where migration to the ocean might occur in the future and determining groundwater recharge balances, as well as assessing other physical/geological features of Amchitka near the test sites. The Science Plan is needed to address the confusing and conflicting information available to the public about radionuclide risks from underground nuclear blasts in the late 1960s and early 1970s, as well as the potential for volcanic or seismic activity to disrupt shot cavities or accelerate migration of radionuclides into the sea. Developing a Science Plan involved agreement among regulators and other stakeholders, assignment of the task to the Consortium for Risk Evaluation with Stakeholder Participation, and development of a consensus Science Plan that dealt with contentious scientific issues. Involvement of the regulators (State of Alaska), resource trustees (U S Fish and Wildlife Service), representatives of the Aleut and Pribilof Island communities, and other stakeholders was essential for plan development and approval, although this created tensions because of the different objectives of each group. The complicated process of developing a Science Plan involved iterations and interactions with multiple agencies and organizations, scientists in several disciplines, regulators, and the participation of Aleut people in their home communities, as well as the general public. The importance of including all parties in all phases of the development of the Science Plan was critical to its acceptance by a broad range of regulators, agencies, resource trustees, Aleutian/Pribilof communities, and other stakeholders. PMID- 15886956 TI - Establishing a context for river rehabilitation, North Fork Gunnison River, Colorado. AB - Initial river rehabilitation efforts along the North Fork Gunnison River in Colorado focused on the use of in-stream structures and channel stabilization to create a single-thread channel with pools along a braided river. These efforts were based on the assumption that the river's braided planform results primarily from land use during the past century. In order to establish a context for further rehabilitation, we evaluated the possibility that the river might be braided as a result of processes independent of land use. We estimated volume, grain-size distribution, and lithology of sediment sources along the river corridor and evaluated the planform stability of the river during the past century using historical sources, aerial photographs covering 1939-1997, and comparison of bankfull discharge and gradient in the study area to values published for braided and meandering rivers. Our results indicate that the North Fork Gunnison River has been primarily braided in its lower reaches during the past few hundred years, although the channel planform tends toward a single thread channel during decades of lower precipitation and discharge. Although land use is not the primary cause of braiding along the North Fork Gunnison River, it has decreased channel stability, and rehabilitation efforts should be designed to reduce these effects. Our results illustrate the importance of planning river rehabilitation measures within a historical context that accounts for both catchment-scale and reach-scale controls on channel processes and planform. PMID- 15886960 TI - Three-dimensional muscular architecture of the human tongue determined in vivo with diffusion tensor magnetic resonance imaging. AB - The myoarchitecture of the tongue is believed to consist of a complex network of interwoven fibers, which function together to produce a near limitless array of functional deformations. These deformations contribute mechanically to speech production and to oral cavity food handling during swallowing. We have previously imaged the 3D myoarchitecture of the mammalian tongue in excised tissue with diffusion tensor MRI, a technique which derives the 3D orientation of intramural fibers as a function of the extent to which a direction-specific MR signal attenuates under diffusion-encoding magnetic gradients. The resulting 3D diffusion tensor defines the relative orientations of the myofiber populations within a region of tissue. In this study, we have extended the use of this method to assess lingual myoarchitecture in normal human subjects in vivo. Subjects were imaged using a diffusion-sensitive stimulated-echo pulse sequence with single shot echo-planar spatial encoding in the midsagittal plane. Differences in lingual fiber orientation were manifested by graduated changes in fiber direction throughout the tissue, without clear anatomical demarcations between regions of the tissue. The anterior tissue was composed generally of orthogonally oriented fibers surrounded by an axially oriented ring of tissue, whereas the posterior portion of the tissue was composed mostly of fibers projecting in the superior and posterior directions. The bulk of the tissue displayed a highly homogeneous, vertically oriented set of fibers, including the anteroinferior region of the tissue and extending nearly to the superior surface. Further analysis of the tissue in terms of diffusion anisotropy demonstrated that the tissue could be represented by varying degrees of anisotropy, with a tendency toward high anisotropy in the dorsal and anteroventral periphery and low anisotropy in the central region of the tissue. These findings demonstrate that the muscular anatomy of the tongue can be displayed as a continuous array of structural units, or tensors, representing fibers of varying orientations throughout the tissue. PMID- 15886961 TI - An anatomical protrusion exists on the posterior hypopharyngeal wall in some elderly cadavers. AB - Dysfunction of the cricopharyngeus is a common cause for dysphagia in the elderly and results in the appearance of the cricopharyngeal bar on a barium swallow radiological examination. However, manometrically normal relaxation of the cricopharyngeus has been observed in patients with a permanent cricopharyngeal bar, which implies that some structural changes may occur in the pharyngoesophageal wall. The aim of this study was to examine the macro microscopic morphological features in the posterior hypopharyngeal wall of elderly human cadavers. Using dissection, histology, and E12 sheet plastination methods, we examined 31 human cadavers (15 females and 16 males; age 60-97 years with a mean of 77 years). We found that about one-third of the cadavers (29%, 9 out of 31) had an anatomical cricopharyngeal protrusion on the posterior hypopharyngeal wall. The protrusion presented two different appearances: a transverse ridge (2 females and 4 males, age 60-91 years) and a tongue-like fold (1 female and 2 males, age 71-86 years). The existence of the anatomical protrusion in the elderly cadaver suggests that such a structural change may become a physical barrier to affect the normal deglutition in the living elderly and should be carefully considered when interpreting radiological and manometrical examinations and with the management of dysphagia. PMID- 15886962 TI - Preswallowing dystonia. AB - A patient is described with some dystonic features related to the preswallowing period of oropharyngeal swallowing in the muscles of deglutition sequentially. This case may be called a "preswallowing dystonia." PMID- 15886963 TI - Reversal of laryngotracheal separation: a detailed case report with long-term followup. AB - Chronic aspiration is a difficult and potentially lethal problem. Patients who have persistent soilage of the upper respiratory tract despite discontinuing oral intake may be offered surgical intervention to avoid life-threatening pulmonary infections. The Lindeman procedures (tracheoesophageal diversion and laryngotracheal separation) have gained popularity as surgical treatments for intractable aspiration because of their efficacy in preventing aspiration and their technical simplicity. A major downside of these procedures is the necessity for a tracheostoma and the loss of speech following surgery. Rarely, patients recover from the neurologic deficits which led to their intractable aspiration and desire reversal of their Lindeman procedure. While few "successful" reversals have been reported, detailed accounts of the long-term results of such patients are lacking. We describe a patient who underwent a laryngotracheal separation for intractable aspiration following a brainstem stroke. In the following six months he experienced significant neurologic recovery and, after careful evaluation, underwent surgical restoration of laryngotracheal continuity. Five years later he speaks fluently and has no dietary restrictions. Videofluooroscopic examination and quantitative voice analysis reveal near-normal laryngeal function. PMID- 15886964 TI - Findings of videofluoroscopic swallowing studies are associated with tube feeding dependency at discharge in stroke patients with dysphagia. AB - The factors associated with tube feeding dependency in stroke patients with dysphagia have received little attention. This study aimed to identify factors independently associated with tube feeding dependency at discharge in stroke patients with dysphagia using videofluoroscopic swallowing studies (VSS) and clinical findings. One hundred eighty-one consecutive tube-fed inpatients with stroke-related dysphagia referred for VSS were enrolled in the study. Of them, 47 patients continued to require tube feeding at discharge. In the final logistic regression analysis model, advanced age, recurrent stroke, confinement to a wheelchair at discharge, long duration from stroke onset to VSS, and stasis in valleculae or pyriform sinuses and aspiration on VSS were independently associated with tube feeding dependency at discharge. This study confirms the association between findings of VSS and tube feeding dependency at discharge in stroke patients with dysphagia. Based on the final regression model, a preliminary equation was proposed to help clinicians better predict the tube feeding dependency at discharge in stroke patients with dysphagia. PMID- 15886965 TI - Change in palatoglossus muscle activity in relation to swallowing volume during the transition from the oral phase to the pharyngeal phase. AB - The purpose of this study was to examine whether the palatoglossus (PG) muscle is involved in the regulation of function during the transition from the oral to the pharyngeal phase. Seven normal adults participated in the study. Smoothed electromyography (EMG) signals of the PG muscle and levator veli palatini (LVP) muscle were collected. Each subject swallowed water at five different volumes: 12.5%, 25%, 50%, 100%, and 150% (or 200%) of his/her optimum swallowing volume. PG muscle waveform showed two patterns of activity: one of a single peak and the other of two peaks. There was no significant difference (p < 0.01) in the timing of emergence between the single peak and the second peak of the two-peak pattern. There were two patterns of PG muscle activity in response to a change in swallowing volume, i.e., one was a pattern in which the activity was correlated to the change in swallowing volume, the other was a pattern in which the activity was not changed but almost at the maximum activity level, irrespective of swallowing volume. We conclude that the PG muscle could be involved in the regulation of swallowing from the oral to the pharyngeal phase. The activity could be influenced by swallowing volume. PMID- 15886966 TI - Clinical, radiographic, and manometric evolution of esophageal involvement by Chagas' disease. AB - The complete manifestation of esophageal Chagas' disease includes nonperistaltic contractions in the esophageal body, absent lower esophageal sphincter (LES) relaxation, and dilatation of the organ. However, some patients have a minor degree of esophageal denervation and esophageal motility that does not imply a diagnosis of achalasia. Our objective was to evaluate the evolution of esophageal involvement by Chagas' disease in 28 patients with dysphagia for solids and liquids and a positive serologic test for the disease, 14 with complete LES relaxation, 4 with partial (incomplete) LES relaxation, and 10 with absent LES relaxation; only 2 of them had mild dilatation. The patients (21 women), aged 43 74 years (median 60 years), were evaluated by clinical, radiographic, and manometric methods that were repeated 3-14 years (median 7 years) later. Dysphagia improved in 13 (46.4%) patients, was worse in 5 (17.9%), and did not change in 10 (35.7%). The radiographic examination did not change in 24 (85.7%) and was worse in 3 (10.7%). Esophageal manometry revealed a change from peristaltic to simultaneous contractions in 2 patients (7.1%), LES relaxation changed from complete to partial in 5 (17.9%), and from partial to absent in 2 (7.1%). There was no further clinical, radiographic, or manometric impairment in 15 (53.6%) patients. The symptom duration was longer and the age when they were evaluated was older in patients with no progression of the disease. We conclude that a conspicuous part of this group of patients with esophageal Chagas' disease and dysphagia had no progression of esophageal disease after 3-14 years. This possibility should be considered when making therapeutic decisions. PMID- 15886967 TI - Tube feeding in infancy: implications for the development of normal eating and drinking skills. AB - Tube feeding is commonly used as a method of giving children nutrition while they are being treated for disease. While this is an effective way of ensuring a child thrives and grows, research studies and clinical experience have shown that long term oral feeding difficulties often arise when the child no longer requires tube feeding. This article gives a critical review of the literature on tube feeding and its effect on normal eating and drinking skills. While few studies have followed a rigorous research design, there is enough literature to identify a number of factors which may be implicated in later feeding difficulties and which therefore need further exploration in research studies. These factors include age at which oral feeding commences, medical complications, exposure to taste and textures during sensitive periods, aversive experiences, and different methods of delivering tube feeds. PMID- 15886970 TI - Ventricular restoration--a surgical approach to reverse ventricular remodeling. AB - Congestive heart failure is most often caused by scar from coronary occlusion. The transition from occluded vessel to scar to dilation results in a remodeled ventricle that changes shape from ellipse to sphere. This shape change following an index event is called remodeling and a surgical approach for restoration (bring back to normal) will be described that uses the patient's own tissue, rather than employing heart replacement by mechanical devices or transplantation. The surgical restoration approach was taken by the RESTORE group that comprises an international medical and surgical team that will report (a) the remodeling infrastructure, role of compensatory remote muscle, and factors underlying surgical restoration decisions, (b) structural basis for ventricular geometric changes and surgical background for restoration, (c) individual rebuilding experience in 1150 patients over 20 years from one center, (d) integrated 5 year results from the RESTORE team in 1198 patients, (e) electrical aspects of restoration in 382 patients with only one AICD used, (f) how restoration improves mechanical synchrony without electrical devices, (g) geometric reasons for secondary mitral insufficiency and impact of adding mitral repair during SVR procedures, and (h) importance of defining site specific scar in no ischemic disease to identify a similar trigger lesion in non ischemic cardiomyopathy. The importance of a team approach by the RESTORE group may set the benchmark for collaborative world wide groups, and thereby depart from traditional focal approaches by individual disciplines. PMID- 15886972 TI - Ventricular structure and surgical history. AB - The Reviews report will establish guidelines that underlie why the "restoration concept" may develop a paradigm shift in thinking, by addressing the geometric underpinnings of heart failure and their evolution. This presentation will (a) show that the underlying structure of the failing dilated heart involves a cardiac architectural change from the normal elliptical shape toward a dilated spherical form, (b) define the anatomic framework of this shape change, (c) convey the functional characteristics of heart function that result from this architectural underpinning, (d) describe the pattern of CHF development, (e) indicate imaging measurement guidelines to follow as heart form adversely changes from ellipse to sphere, (f) identify how such architectural changes alter prognosis, and (g) develop a historical evolution of surgical approaches to alter form to improve function to create the background for subsequent RESTORE team reports of current restoration to treat CHF and its complications. PMID- 15886971 TI - Heart failure following anterior myocardial infarction: an indication for ventricular restoration, a surgical method to reverse post-infarction remodeling. AB - Anterior myocardial infarction produces abrupt left ventricular (LV) dysynergy and global systolic dysfunction. Rapid intense neurohumoral activation, infarct expansion, and early ventricular chamber dilatation all contribute to restoring a normal stroke volume despite a persistently depressed ejection fraction. Continued neurohumoral activation provokes late remodeling of the remote non infarcted myocardium, characterized by an abnormal progressively increasing LV volume/mass ratio that leads to further LV remodeling. Heart failure is a progressive disorder of LV remodeling. Heart failure from post-infarction remodeling is unique because of the persistent non-functioning scar that self- perpetuates abnormal loading conditions and neurohumoral activation. Medical therapy attenuates remodeling and improves survival but does not change the size of the scar. Surgical ventricular restoration to exclude the non-functioning infarct from the ventricular cavity decreases ventricular volumes, increases global ejection fraction, attenuates neurohumoral activation and yields an excellent 5-year survival. Combined medical and surgical therapy is recommended in this patient population. PMID- 15886973 TI - Endoventricular patch reconstruction of ischemic failing ventricle. a single center with 20 years experience. advantages of magnetic resonance imaging assessment. AB - The left ventricular reconstruction (LVR) with endoventricular circular patch plasty (EVCPP) was reported in 1984 as a surgical method to rebuild left ventricular aneurysm or asynergy after myocardial infarction. Scarred LV wall can be dyskinetic or akinetic according to the type of infarction (transmural or not), and the progressive dilatation of LV (remodeling) depends on the size of the asynergic scar. Assessment of this extension and of LV volume and performances, is easy and reliable by magnetic resonance (CMR). The surgical technique is based on the insertion inside the ventricle on contractile myocardium, of a circular patch restoring curvature and physiological volume, and allowing exclusion of asynergic non resectable regions. The ventricular reconstruction method also has other components that include coronary revascularization (almost always), mitral repair (if needed) and endocardectomy when spontaneous or inducible ventricular tachycardia (VT) are present. The experience of the authors (> 1100 cases) and results obtained by other Centers, allows proposal of this technique as a way to treat the ischemic failing ventricle. PMID- 15886974 TI - Surgical ventricular restoration: the RESTORE Group experience. AB - Congestive heart failure may be caused by late left ventricular (LV) dilation following anterior infarction. Early reperfusion prevents transmural necrosis, and makes the infarcted segment akinetic rather than dyskinetic. Surgical ventricular restoration (SVR) reduces LV volume and creates a more elliptical chamber by excluding scar in either akinetic or dyskinetic segments. The international RESTORE group applied SVR in a registry of 1198 post-infarction patients between 1998 and 2003. Early and late outcomes were examined and risk factors identified.Concomitant procedures included coronary artery bypass grafting in 95%, mitral valve repair in 22%, and mitral valve replacement in 1%. Overall 30-day mortality after SVR was 5.3% (8.7% with mitral repair vs. 4.0% without repair, p < .001) Perioperative mechanical support was uncommon (< 9%). Global systolic function improved postoperatively, as ejection fraction increased from 29.6 +/- 11.0% to 39.5 +/- 12.3% (p < .001) and left ventricular end systolic volume index decreased from 80.4 +/- 51.4 ml/m(2) to 56.6 +/- 34.3 ml/m(2) (p < .001). Overall 5-year survival was 68.6 +/- 2.8%, Logistic regression analysis identified EF < or = 30%, LVESVI > o = 80 ml/m(2), advanced NYHA functional class, and age > or =75 years as risk factors for death. Five year freedom from hospital readmission for CHF was 78%. Preoperatively, 67% of patients were class III or IV, and postoperatively 85% were class I or II.SVR improves ventricular function and is highly effective therapy in the treatment of ischemic cardiomyopathy with excellent 5-year outcome. PMID- 15886975 TI - Ventricular arrhythmias after LV remodelling: surgical ventricular restoration or ICD? AB - Ventricular arrhythmias cause ~50% of deaths in remodeled ventricles after myocardial infarction, and the Multicenter Automatic Defibrillator Implantation Trial (MADIT II) showed that the Implantable Cardioverter Defibrillator (ICD) saved lives in high risk coronary patients with advanced left ventricular dysfunction. We studied 382 patients with remodeled hearts by preoperative Ventricular stimulation (PVS) to evaluate surgical ventricular restoration (SVR) that excludes scar and lower ventricular volume alters the early and late arrhythmia process without ICD utilization. METHODS: Clinical and hemodynamic results before and after SVR in post-infarction patients, are compared to contrast spontaneous and/or inducible ventricular tachycardia to patients without arrhythmias. Study arrhythmia groups included: Spontaneous in 87 patients with clinical documented ventricular arrhythmias and inducible or not inducible ventricular tachycardia: Inducible in 105 patients without clinical ventricular arrhythmias but PVS inducible ventricular tachycardia; and No arrhythmias in 190 patients without spontaneous or PVS inducible ventricular tachycardia. RESULTS: Preoperative LV end systolic volume index helped define preoperative arrythmia potential: Spontaneous > 120/m(2), inducible > 100 ml/m(2), and none < 100ml/m(2). Overall operative mortality rate was 7.6% (29/382). Sudden cardiac death rate was 2.5% causing 18.7% of all deaths. Surgical management reduced inducible ventricular tachycardia, from 41% preoperatively (144/352) to 8% (26/307) at early study, and 8% (14/177) one year later. Cardiac mortality was low at 5-years and not different between groups, despite use of only one late ICD device. CONCLUSIONS: Favorable electrical success rate and low mortality always included volume reduction to interrupt functional re-entry circuits, but also added endocardiectomy, cryoablation, CABG and mitral repair when needed. Overall SVR findings show volume and shape alteration limits ventricular arrhythmias that impair prognosis, and suggests ICD devices are not needed. PMID- 15886976 TI - Mechanical synchrony: role of surgical ventricular restoration in correcting LV dyssynchrony during chamber rebuilding. AB - Cardiac failure is frequently complicated by intra and or interventricular conduction delay that results in dyssynchronized cardiac contraction and relaxation. In contrast to an electrical intervention by biventricular pacing, this study tests the capacity of geometric rebuilding by surgical ventricular restoration (SVR) to restore a more synchronous contractile pattern through mechanical reconstruction without exogenous pacing input. Thirty patients (58 +/- 8 years) undergoing SVR at the Cardiothoracic Center of Monaco were prospectively evaluated with a protocol which uses simultaneous measurements of ventricular volumes and pressure to construct pressure/volume (P/V) and pressure/length (P/L) loops. Mean QRS duration was within normal limits (100 +/- 17 ms) preoperatively. Preoperative LV contraction was highly asynchronous. Endocardial time motion was either early or delayed at the end-systolic phase, yielding P/L loops with abnormal in size, shape, and orientation. Postoperatively, SVR resulted in leftward shifting of P/V loops and increased area; endocardial time motion and P/L loops almost normalized. The hemodynamic consequences of SVR included improved ejection fraction; reduced end-diastolic and end-systolic volume index; more rapid peak filling rate; peak ejection rate and mechanical efficiency resulting in mechanical intraventricular resynchronization that improves LV performance. PMID- 15886977 TI - Functional ischemic mitral regurgitation in anterior ventricular remodeling: results of surgical ventricular restoration with and without mitral repair. AB - Ischemic functional mitral regurgitation following ischemic cardiomyopathy is a secondary phenomenon to ventricular dilation, and therapeutic approaches to this complication are not uniform. Solutions to improve mitral function include either mitral repair or observing the effects of coronary revascularization and/or ventricular rebuilding during surgical ventricular restoration (SVR). The present study of 108 patients (comprising 18% of our 588 SVR population) reports the effects of mitral repair following SVR and CABG by comparing geometric, functional, hemodynamic and outcome changes to SVR patients without mitral repair. The degree of mitral regurgitation went from 2.9 +/- 1.2 before to 0.7 +/ 0.7 after SVR and mitral repair. SVR improved EF from 29 +/- 7% to 34 +/- 10% p 0.001; reduced end diastolic volume from 243 +/- 74 to 163 +/- 53 ml and end systolic volume from 170 +/- 63 to 107 +/- 41 ml, p 0.000. Ventricular size and shape geometric measurements improved in all patients, either with and without mitral repair. SVR improved tenting and papillary muscle width between muscle heads in all patients, but alterations in mitral annular size improved only following mitral repair. Preoperative mitral regurgitation occurred in patients with larger ventricular volume and lower ejection fraction and was an independent predictor of operative mortality risk. PMID- 15886978 TI - Role of site selection for left ventriculoplasty to treat idiopathic dilated cardiomyopathy. AB - Ventriculoplasty was introduced to treat patients with chronic progressive heart failure from end-stage non ischemic dilated cardiomyopathy, which was presumed to be a homogeneous disease. However ventriculoplasty is not commonly used today, because variable results follow using only lateral ventriculoplasty as the treatment tool. This report traces our evolution in surgical management, defines that the homogeneous disease concept is not correct, and centers upon the importance of site selection, appropriate timing, and methods of patient selection. Left ventriculoplasty by either partial left ventriculoplasty (PLV) or septal anterior ventricular exclusion (SAVE or Pacopexy) was performed in 96 high risk (44% NYHA Class III, and 56% Class IV) patients with idiopathic dilated cardiomyopathy over the past 8 years. Overall hospital mortality was 8% in elective operations and 57% in emergency operations. Hemodynamic and functional improvement was evident from ejection fraction rising from 20% to 31%, and improved NYHA Class 3.6 to 1.8. The global series showed 1, 5 and 7 year survival rate was 66.4, 44.7 and 41.3%, respectively. However surgical results improved as experience allowed development of a strategy for timing, and defining proper exclusion site selection. Non homogeneous ventricular disease was identified, and 69% 4 year survival resulted from our intraoperative definition of the most diseased segment (septum or lateral wall), and then exclusion of this site. This evolving experience indicates that left ventriculoplasty is effective treatment for patients with end-stage cardiomyopathy, provided proper patient selection, appropriate timing of the operation, and choice of procedure are used as keys to a successful outcome. PMID- 15886979 TI - Neurohormonal intervention to reduce sudden cardiac death in heart failure: what is the optimal pharmacologic strategy? AB - Sudden cardiac death (SCD) accounts for up to 50% of deaths in patients with heart failure (HF), depending on severity of symptomatic impairment and left ventricular dysfunction. Neurohormonal therapy directed at the renin-angiotensin aldosterone system may reduce the propensity to SCD through improved hemodynamic responsiveness, reduced sympathetic tone in the myocardium and inhibition of cardiac remodelling. Angiotensin converting enzyme (ACE) inhibitors reduce overall mortality in chronic HF, the greatest benefit appearing to arises from reduction of HF progression rather than SCD. In HF patients who experience myocardial infarction (MI) reduced incidence in SCD may make a more marked contribution to the mortality benefits of ACE inhibition. Addition of beta blocker therapy to ACE inhibition has consistently resulted in a reduction in SCD in patients with either mild-to-moderate or severe HF, and in the presence or absence of MI; the reduction in SCD is of the order of one-third versus placebo. Aldosterone blockade reduces the risk of SCD in advanced chronic heart failure (when added to ACE inhibitor) and in HF associated with acute MI (when given in addition to both ACE inhibitor and beta blocker). The evidence base suggests that for maximal SCD risk reduction in HF, beta-blocker therapy is advisable in combination with standard ACE inhibitor therapy, with addition of aldosterone blockade to this regimen for particular groups of heart failure patients. PMID- 15886980 TI - Sudden cardiac death after heart failure; symptomatic versus disease treatment? An editorial comment. PMID- 15886981 TI - Visibility of the origin and proximal course of coronary arteries on non-ECG gated heart CT in patients with congenital heart disease. AB - BACKGROUND: There is little information on the ability of non-ECG-gated cardiac CT to demonstrate the coronary arteries of children. OBJECTIVE: To evaluate the visibility of the origin and proximal course of coronary arteries on non-ECG gated cardiac CT, in which the coronary artery was not of primary diagnostic concern, in children with congenital heart disease. MATERIALS AND METHODS: From December 2002 to March 2004, 126 cardiac CT examinations from 104 children (median age 11 months; age range 1 day to 15 years) were evaluated. All patients had ventriculo-arterial concordance and no malformations of the great arteries; those with coronary artery anomalies were excluded. Contrast-enhanced 16-slice spiral CT was performed without ECG-gating and multiplanar images for coronary arteries were obtained. The visibility of coronary artery origins was graded on a three-point scale, while nine segments of the arteries were graded on a four point scale. CT images in which it was possible to trace the coronary arteries were considered diagnostic. The visibility of each whole coronary artery and the origins and proximal four segments of coronary arteries were calculated. The visibility of coronary arteries was also correlated with patient age. RESULTS: The percentage of CT images of diagnostic quality was 49.3% for the whole coronary artery and 81.7% for the origins and proximal four segments. There was a significant positive correlation between the visibility of coronary arteries and age. CONCLUSIONS: Non-ECG-gated cardiac CT, in which the coronary artery is not of primary diagnostic concern, is frequently able to visualize the origin and proximal course of coronary arteries and may be helpful in detecting coronary artery anomalies in children with congenital heart disease. PMID- 15886982 TI - Rare localization of osteoid osteoma in the patella. PMID- 15886983 TI - Childhood atypical meningioma with perineural spread: MR findings. AB - Meningiomas are uncommon in children. When they occur, they are frequently associated with neurofibromatosis type 2. Childhood meningiomas are generally large and commonly associated with cyst formation and an unusual location. Perineural tumor spread, occasionally associated with head and neck malignancies, is very rare in meningiomas. We present the MR findings of an atypical meningioma with perineural spread in a 4.5-year-old girl. PMID- 15886984 TI - Compensatory hypertrophy of renal parenchyma presenting as a mass lesion. PMID- 15886985 TI - Elastic vibrations in seamless microtubules. AB - Parameters characterizing elastic properties of microtubules, measured in several recent experiments, reflect an anisotropic character. We describe the microscopic dynamical properties of microtubules using a discrete model based on an appropriate lattice of dimers. Adopting a harmonic approximation for the dimer dimer interactions and estimating the lattice elastic constants, we make predictions regarding vibration dispersion relations and vibration propagation velocities. Vibration frequencies and velocities are expressed as functions of the elastic constants and of the geometrical characteristics of the microtubules. We show that vibrations which propagate along the protofilament do so significantly faster than those along the helix. PMID- 15886986 TI - The influence of a cellular size distribution on NMR diffusion measurements. AB - In the magnetic resonance community, the usefulness of diffusion-weighted imaging is undisputed. However, the correct interpretation of multicomponent diffusion is not widely agreed upon, and progress in this direction is impeded by several confounding experimental results. It is thus of great importance to resolve possible interfering factors. The objective of the present study is to examine the influence of a cellular size distribution. Using the Karger equations, numerical calculations show that even substantial cellular size variations induce only small changes in the estimated compartmental diffusion constants and volume fractions. PMID- 15886987 TI - [Comparison of several viral vectors for gene therapy of corneal endothelial cells]. AB - AIM: In this paper we compare the transduction efficiency, toxicity, and safety of retroviral vectors [equine infectious anemia virus (EIAV), human immunodeficiency virus-1 (HIV-1), human foamy virus (PFV] and adenovirus (Ad) for potential use in gene therapy of corneal endothelial cells. METHOD: Murine corneal endothelial cells were transduced with EIAV, HIV-1, PFV, and Ad, resulting in the overexpression of a green fluorescent protein (eGFP) transgene marker. The transduction efficiency was assessed by flow cytometry, while cytotoxicity and apoptosis rate were detected by annexin V/propidium iodide (PI) stain. RESULTS: Ad had the highest transduction efficiency with 99% of the cells expressing the transgene, followed by EIAV (95%), HIV-1 (75%), and PFV (43%). However, the high transduction efficiency of Ad also resulted in the highest apoptosis rate (25%) in the corneal endothelial cells. There was no detectable difference in the toxicity between PFV and HIV-1 (10%). EIAV transduction had the lowest cytotoxicity, with only 3% of the cells being annexin V/PI positive. CONCLUSION: Compared to other vectors EIAV exhibited high transduction efficiency combined with low toxicity to corneal endothelial cells. Therefore, it is a powerful tool for gene therapy applications in selected corneal endothelial diseases. PMID- 15886988 TI - [Implantation of lid weights for therapy of lagophthalmos]. AB - BACKGROUND: The aim of this study was to compare the clinical outcome and complication rate of rigid gold implants and flexible platinum chains in 96 patients treated for lagophthalmos. PATIENTS AND METHODS: We treated 50 patients with peripheral facial palsy and lagophthalmos using rigid gold implants and 46 using a flexible platinum chain. Besides subjective assessment by the patients, reduction of lagophthalmos, improvement of visual acuity, and pre- and postoperative grade of keratopathy were evaluated. RESULTS: Both groups showed a statistically significant reduction of lagophthalmos and keratopathy and increase of visual acuity. Postoperative complications were observed in both groups. Two extrusions occurred in the group receiving gold implants. The incidence of corneal astigmatism and bulging of the implant was statistically significantly lower in the platinum chain group. CONCLUSIONS: The use of flexible platinum chains instead of rigid gold implants for therapy of lagophthalmos leads to a reduction of the complication rate, thus improving the clinical outcome. PMID- 15886989 TI - [Cataract patients with pseudoexfoliation (PEX) syndrome among a population with high prevalence of PEX. Clinical findings and morphological and surgical characteristics]. AB - BACKGROUND: Cataracts are the most frequent treatable cause of noninfectious blindness in Oman. Therefore, in 2002 a study was performed at our department of ophthalmology to evaluate cataract patients for pseudoexfoliation. METHODS: In a prospective comparative cohort study, 370 age-related cataract eyes out of an evaluable population of 498 cataracts were evaluated by biomicroscopy, confocal corneal analysis, and electron microscopy and followed up after 6 months. RESULTS: Of 370 age-related cataracts without glaucomas 171 had PEX. Preoperative manifestations were 89 subluxations, 15 vitreous prolapses, 6 ciliolenticular blocks, and 7 intravitreal dislocations. In 40 Omani cataract eyes without PEX the endothelial cell count was normal compared to the age-matched US norm and corneal thickness was less than Caucasians, but more than Afro-Americans. CONCLUSIONS: This study reports on a high incidence and frequent preoperative complications (68%) of the surgical cataract cases in an Arabic population. The development of serious complications and blindness is related to the duration of the disease and the lack of timely surgery. PMID- 15886990 TI - Magnetic orientation and magnetoreception in birds and other animals. AB - Animals use the geomagnetic field in many ways: the magnetic vector provides a compass; magnetic intensity and/or inclination play a role as a component of the navigational 'map', and magnetic conditions of certain regions act as 'sign posts' or triggers, eliciting specific responses. A magnetic compass is widespread among animals, magnetic navigation is indicated e.g. in birds, marine turtles and spiny lobsters and the use of magnetic 'sign posts' has been described for birds and marine turtles. For magnetoreception, two hypotheses are currently discussed, one proposing a chemical compass based on a radical pair mechanism, the other postulating processes involving magnetite particles. The available evidence suggests that birds use both mechanisms, with the radical pair mechanism in the right eye providing directional information and a magnetite based mechanism in the upper beak providing information on position as component of the 'map'. Behavioral data from other animals indicate a light-dependent compass probably based on a radical pair mechanism in amphibians and a possibly magnetite-based mechanism in mammals. Histological and electrophysiological data suggest a magnetite-based mechanism in the nasal cavities of salmonid fish. Little is known about the parts of the brain where the respective information is processed. PMID- 15886991 TI - Optical quality changes of the ocular lens during induced parr-to-smolt metamorphosis in Rainbow Trout (Oncorhynchus mykiss). Ocular lens optical quality during induced salmonid metamorphosis. AB - The effect of an induced salmonid parr-to-smolt metamorphosis ('smoltification') on the optical quality of the ocular lens was studied. In two separate experiments, rainbow trout (Oncorhynchus mykiss) parr were fed thyroxine in their diet to induce the metamorphosis. Lenses were excised at regular samplings during the treatment period and optically scanned using a custom scanning laser monitor. Radioimmunoassay was used to measure serum titers of thyroxine and 3,5,3'-triiodo L: -thyronine. It was found that lens optical quality was consistently negatively correlated with 3,5,3'-triiodo-L: -thyronine levels, but not with thyroxine levels. To test if thyroid hormones are directly responsible for the change in optical quality, rainbow trout lenses were cultured for 72 h in a medium containing 3,5,3'-triiodo-L: -thyronine, but no effect was observed. The significance of these findings in the contexts of the fishes' visual capabilities and smolting physiology is discussed. PMID- 15886992 TI - Sensory ecology of predator-prey interactions: responses of the AN2 interneuron in the field cricket, Teleogryllus oceanicus to the echolocation calls of sympatric bats. AB - We observed the responses of the AN2 interneuron in the Pacific field cricket, Teleogryllus oceanicus, a cell implicated in eliciting avoidance flight away from bats, to acoustic stimuli representing the echolocation calls of bats as well as field recordings of search and gleaning attack calls of six species of insectivorous sympatric bats (West Australia, Australia: Tadarida australis, Chalinolobus goudii, Nyctophilus geoffroyi; Queensland, Australia: Vespadelus pumilus, Myotis adversus; Kaua'i, Hawai'i: Lasiurus cinereus). The broad frequency sensitivity of the AN2 cell indicates that T. oceanicus has evolved to detect a wide range of echolocation call frequencies. The reduced sensitivity of this cell at frequencies higher than 70 kHz suggests that some bats (e.g., the gleaning species, N. geoffroyi) may circumvent this insect's auditory defences by using frequency-mismatched (allotonic) calls. The calls of the freetail bat, T. australis evoked the strongest response in the AN2 cell but, ironically, this may allow this bat to prey upon T. oceanicus as previous studies report that under certain conditions, flying crickets exhibit ambiguous directional responses towards frequencies similar to those emitted by this bat. Short duration calls (1 -2 ms) are sufficient to evoke AN2 responses with instantaneous spike periods capable of causing defensive flight behaviours; most bats tested emit calls of durations greater than this. The short calls of N. geoffroyi produced during gleaning attacks may reduce this species' acoustic conspicuousness to this cricket. PMID- 15886993 TI - Differential stress responses in fish from areas of high- and low-predation pressure. AB - We subjected fish from regions of high and low levels of predation pressure in four independent streams to a mild stressor and recorded their opercular beat rates. Fish from low-predation areas showed higher maximum, minimum and mean opercular beat frequencies than fish from high-predation regions. The change in opercular beat frequency (scope) was also significantly greater in fish from low- than in fish from high-predation regions. Under normal activity levels, however, low predation fish showed a reduced opercular beat frequency, which may be indicative of reduced activity levels or metabolic rate. Opercular beat frequency was negatively correlated with standard length as one would expect based on higher metabolic rates in smaller fish. We suggest that these contrasting stress responses are most likely the result of differential exposure to predators in fish from high- and low-predation areas. We argue that reduced stress responses in high-predation areas evolved to prevent excessive energy expenditure by modulating the fright response. PMID- 15886994 TI - Stereological studies of capillary length density in the frontal cortex of schizophrenics. AB - The presence of microvasculature abnormalities in the prefrontal cortex of schizophrenics was proposed in a recent study of molecular signatures of schizophrenia [Prabakaran et al (2004) Mol Psychiat 9:684-697]. To assess this possibility further, we investigated capillary length densities in prefrontal cortex area 9 and anterior cingulate cortex area 24 in postmortem brains from 13 schizophrenics and 13 age- and sex-matched controls. To check that our sample of brains shared cardinal neuropathological features of schizophrenia with previously reported case studies, we also measured cortical gray matter volumes and cortical thickness in areas 9 and 24. The mean cortical gray matter volume was significantly reduced in brains from schizophrenics compared to controls. Mean cortical thickness was significantly reduced in area 24, but not in area 9, in schizophrenics. There were no differences in mean capillary length densities in either area 9 or 24 between the two groups. Thus, alterations in capillary length density in the prefrontal cortex cannot be considered a general feature of schizophrenia. Compromised brain metabolism and occurrence of oxidative stress in the brain of schizophrenics are likely caused by other mechanisms. PMID- 15886995 TI - Molecular characterization of Blastocystis isolates in the Philippines by riboprinting. AB - Extensive genomic polymorphism has been demonstrated among morphologically identical Blastocystis isolates. To this end, 32 Blastocystis isolates from the Philippines (12 from humans, 12 from pigs and 8 from chickens) were analyzed genetically by riboprinting or restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR) amplified small subunit rDNA. Three distinct riboprint patterns were observed from the HinfI digestion, while four patterns resulted from the RsaI digestion of Blastocystis SSU rDNA. Restriction fragment profiles between Blastocystis isolates from different hosts were generally different from each other. However, Blastocystis isolates within each host group were practically the same. Cluster analysis of the riboprint patterns revealed seven distinct groups of the Blastocystis isolates, including a zoonotic strain. These results demonstrate the genetic heterogeneity of Blastocystis in the Philippines and a support to the idea of the organism's zoonotic potential. PMID- 15886996 TI - The development of human genetics in Germany; a personal view. AB - A personal account is given of the reconstruction and development of human genetics in Germany during the years following World War 2. An important stimulus was funding, as a result of the recognition of the genetic hazards of atomic radiation. Starting from 1960, human genetics institutes were progressively established throughout West Germany; comparable development was later in East Germany because of political factors. The first genetic counselling units were formed in 1972, but molecular biology only became an integral part of human genetics institutes at a relatively late stage. Close international links have characterised post-war human genetics in Germany from the outset and a tradition of close links with developing countries has also been established. PMID- 15886997 TI - Localisation of merosin-positive congenital muscular dystrophy to chromosome 4p16.3. AB - The congenital muscular dystrophies (CMD) are a heterogeneous group of autosomal recessive disorders, which present within the first 6 months of life with hypotonia, muscle weakness and contractures, associated with dystrophic changes on skeletal muscle biopsy. We have previously reported a large consanguineous family segregating merosin-positive congenital muscular dystrophy, in which involvement of known CMD loci was excluded. A genome-wide linkage search of the family conducted using microsatellite markers spaced at 10-Mb intervals failed to identify a disease locus. A second scan using a high-density SNP array, however, permitted a novel CMD locus on 4p16.3 to be identified (multipoint LOD score 3.4). Four additional consanguineous CMD families with a similar phenotype were evaluated for linkage to a 4.14-Mb interval on 4p16.3; however, none showed any evidence of linkage to the region. Our findings further illustrate the utility of highly informative SNP arrays compared with standard panels of microsatellite markers for the mapping of recessive disease loci. PMID- 15886998 TI - Evolution versus constitution: differences in chromosomal inversion. AB - We compared the chromosomal breakpoints of evolutionary conserved and constitutional inversions. Multicolor banding and human-specific bacterial artificial chromosomes were applied to map the breakpoints of constitutional pericentric inversions on human chromosomes 2 and 9. For the first time, we present a high-resolution analysis of the breakpoint regions, which are characterized by gene destitution, co-localization with fragile sites, multitude repeats as well as pseudogenes and, remarkably, a large sequence homology to the opposite breakpoint. In contrast, evolutionary inversion breakpoints lack such extensive cross-hybridizing regions and are often associated with fragile sites of the genome and low-copy repeats. These molecular characteristics gave evidence for different types of inversion formation and indicate that evolutionary inversions cannot originate from constitutional inversions like those of chromosomes 2 and 9. Finally, the constitutional inversion breakpoints were investigated on three different great ape species and on four test persons each bearing the same cytogenetically determined inversion on chromosomes 2 and 9, respectively. Our data indicate the existence of different molecular breakpoints for the two variant chromosomes. PMID- 15886999 TI - A century later Farabee has his mutation. PMID- 15887000 TI - A case of autism and uniparental disomy of chromosome 1. AB - We report a male child with autism found to have maternal uniparental disomy (UPD) of chromosome 1. The child met diagnostic criteria for the three symptom domains of autism: language impairment, deficient social communication and excessively rigid and repetitive behaviours. He also had a variety of features often associated with autism, including mild mental retardation, small head circumference, hyperactivity, poor fine motor skills, slightly dysmorphic facial features and a heightened interest in olfactory stimulation. His brother, who did not have chromosome 1 UPD, was also autistic. The mother, but not the father, had a history of psychiatric illness and a number of personality and social traits similar to the core features of autism. The discovery of the cytogenetic abnormality was made during the course of a genome-wide linkage screen, wherein genotypes at 6 out of 17 chromosome 1 markers were non-Mendelian and all transmissions were consistent with UPD. Further genotyping (a total of 54 markers) revealed alternating regions of heterodisomy and isodisomy. Whereas chromosome 1 UPD has not been shown to cause disease by effects on imprinting, numerous reports exist of the abnormality unmasking recessive disease-causing mutations. In agreement with this, one of the regions of isodisomy overlaps an emerging chromosome 1 region of interest in autism located at 150-160 Mb. PMID- 15887002 TI - Evaluation of primary production in Lake Erie by multiple proxies. AB - Direct measurements of rates of primary production in Lake Erie are few and uncertainties surround rate measurements based on radiocarbon and the light-dark bottle incubation methods. For these reasons, we conducted a series of simultaneous primary productivity measurements in Lake Erie in July and August of 2003, based on incubation with [14C]-NaHCO3, the light-dark bottle method, and incubation with (18)O enriched water. Significant differences in the rates of primary production obtained by incubations with [(18)O]-H2O (0.19-34.60 mmol-O2 m(-3) h(-1)), [14C]-NaHCO3 (0.03-90.50 mmol-C m(-3) h(-1)), and light-dark bottles (0.06-60.78 mmol-O2 m(-3) h(-1)) were evident in six out of nine comparisons. Within the epilimnion, [(18)O]-H2O rates of primary production were significantly different from rates based on [14C]-NaHCO3 and light-dark bottles in all four comparisons and lower rates were obtained in three out of four comparisons. Eutrophic conditions in Sandusky Bay, Lake Erie were evident from the high primary production rates of 20.50-34.60 mmol-O2 m(-3) h(-1) ([(18)O] H2O), 34.39-90.50 mmol-C m(-3) h(-1) ([14C]-NaHCO3), and 46.66-60.78 mmol-O2 m( 3) h(-1) (light-dark bottle). The photosynthetic quotient (PQ), or ratio of O2 production to CO2 consumption during photosynthesis, averaged 0.64+/-0.33 and 1.93+/-1.93, respectively, based on a comparison of [(18)O]-H2O to [14C]-NaHCO3 rates or light-dark bottle to [14C]-NaHCO3 production rates, respectively, demonstrating that photosynthesis in Lake Erie communities primarily follows expected stochiometric trends. The average of the ratio of production rates based on incubation with [(18)O]-H2O relative to those obtained by the light-dark incubation method was 0.66+/-0.33, indicating a tendency for the [(18)O]-H2O method to provide slightly lower estimates of production in Lake Erie. Lower estimates of primary production based on [(18)O]-H2O incubation relative to the other two approaches is most likely a consequence of consumption of labeled O2 within the cell or dilution of label by the release of O2 from supersaturated cells. This latter effect may be particularly characteristic of eutrophic environments. PMID- 15887001 TI - [Treatment and prophylaxis for cluster headaches and other trigeminal autonomic headaches. Revised recommendations of the German Migraine and Headache Society]. AB - Following the new IHS classification, cluster headache, paroxysmal hemicrania, and short-lasting unilateral neuralgiform headache with conjunctival injection and tearing (SUNCT syndrome) are included in the classification as trigeminal autonomic cephalgias (TAC). The similarities of these syndromes suggest a considerable shared pathophysiology. These syndromes have in common that they involve activation of trigeminovascular nociceptive pathways with reflex cranial autonomic activation. Clinically, this physiology predicts pain with some combination of lacrimation, conjunctival injection, nasal congestion, or eyelid edema. Broadly the management of TAC comprises acute and prophylactic treatment. Some types of trigeminal autonomic headaches such as paroxysmal hemicrania and hemicrania continua have, unlike cluster headaches, a very robust response to indomethacin, leading to a consideration of indomethacin-sensitive headaches. This review covers the clinical picture and therapeutic options. Although studies following the criteria of evidence-based medicine (EBM) are rare, most patients can be treated sufficiently. PMID- 15887003 TI - [Cervicogenic head and neck pain]. AB - It is discussed controversially whether cervicogenic pain in the head and/or neck is a pathogenic entity. The good results obtained with manual therapy in patients with head and neck pain contradict the refusal of the majority of the neurologists to accept the diagnosis "cervicogenic headache." Complaints about headache are frequently encountered in the general ENT clinic. In many cases, the diagnosis of the different types of headaches can be based on the anamnesis. It is difficult to define a tension headache, because it is not a sharply defined syndrome and the disturbance of the neck represents only one of many factors. The versatile picture of the cervicogenic headache is caused by the complex neural connections in the region of the upper cervical spine. The differential diagnosis of the cervicogenic headache is described. PMID- 15887004 TI - Effect of dopamine D3 receptor blockade on renal function and glomerular size in diabetic rats. AB - Dopamine D2-like receptors, including D2, D3, and D4 receptors, are involved in the regulation of glomerular hyperfiltration due to diabetes mellitus. These hemodynamic alterations represent a risk factor for the later development of diabetic nephropathy. The aim of the present study was to determine whether the D3 receptor subtype modulates the diabetes-induced increase in glomerular filtration rate (GFR) in rats. Renal function was studied in Sprague-Dawley rats 14 days after induction of a moderate diabetes mellitus (DM) by streptozotocin and in non-diabetic controls (CON). Rats were orally treated either with the peripherally acting, selective dopamine D3 receptor antagonist BSF 135170 (BSF, 10 mg/kg per day for 2 weeks) or with vehicle (VHC). Perfusion-fixed kidneys were used for estimation of glomerular volume. In conscious rats, which were treated with BSF, the DM-induced increase in fluid intake, urinary output, and renal sodium excretion was significantly less pronounced than in the vehicle group (DM VHC). In the clearance experiments, GFR in CON was about 0.84+/-0.04 ml/min per 100 g body weight. The DM-VHC group presented a significant glomerular hyperfiltration (1.09+/-0.04 ml/min per 100 g body weight). Treatment with BSF significantly lowered GFR towards levels of CON. The estimated glomerular volume was 0.73+/-0.03 x 10(6) microm3 in the CON-VHC group and 0.86+/-0.04 x 10(6) microm3 in the DM-VHC animals. Interestingly, treatment with BSF decreased the glomerular volume in both groups. Irrespective of BSF treatment, kidney wet weight related to body weight was about 36% higher in DM animals compared with CON animals. We conclude that dopamine D3 receptors represent a target for the modulation of diabetes-induced glomerular hyperfiltration. Therefore, the results encourage the testing of the possible beneficial effects of long-term D3 receptor blockade on the development of diabetic nephropathy. PMID- 15887005 TI - The timing and intensity of transcranial magnetic stimulation, and the scalp site stimulated, as variables influencing motor sequence performance in healthy subjects. AB - OBJECTIVE: The increasing therapeutic use of transcranial magnetic stimulation (TMS) in disorders of cortical excitability raises the need for reliable stimulus variables. Stimulation of cortical motor areas influences motor programming and execution. We investigated the effects of TMS delivered over various cortical motor areas during the reaction time (RT) on the execution of sequential rapid arm movements in healthy subjects. METHODS: Subjects performed a five-submovement (S1-S5) motor sequence mainly involving upper limb proximal muscles. RT and movement time (MT) were measured. We delivered late (close to movement onset) and early (close to the go signal) TMS over the primary motor area (M1-FDI hot-spot for the first dorsal interosseus, M1-D hot-spot for the deltoid muscle), the premotor (PM) area, and the supplementary motor area (SMA), using subthreshold and suprathreshold intensity, single and triple pulses. RESULTS: The motor sequence showed a characteristic pattern of submovement duration, S2-S3-S4 being faster than S1 and S5. Late TMS prolonged RT only when high-intensity pulses were delivered over M1-FDI. Single- and triple-pulse TMS over M1-D or M1-FDI significantly prolonged MT with a dose-related effect. Suprathreshold triple pulse TMS over the PM-but not over the SMA-also lengthened the MT but did not change RT. Early triple-pulse TMS reduced the RT independently from the stimulus intensity and scalp site. SMA and PM-but not M1-D-stimulation also reduced the MT. Single-pulse TMS over the SMA, despite being delivered through a double-cone coil, did not change RT or MT. CONCLUSIONS: TMS-induced changes in the kinematics of a sequential arm movement depend closely on the timing of TMS interference, the scalp site stimulated, and the intensity (and number) of stimuli delivered. Late TMS interference inhibits, whereas early interference facilitates, motor performance. The cortical motor region most sensitive to TMS-induced inhibition is that below the scalp site for M1-FDI. In contrast, TMS-induced facilitation has no strict topographic organization. Particularly for MT (although inhibitory and facilitatory effects both depend on stimulation at high intensities) intensity is less crucial than timing of interference and scalp site. PMID- 15887006 TI - Identification of c-Fos immunoreactive brainstem neurons activated during fictive mastication in the rabbit. AB - In the present study we used the expression of the c-Fos-like protein as a "functional marker" to map populations of brainstem neurons involved in the generation of mastication. Experiments were conducted on urethane-anesthetized and paralyzed rabbits. In five animals (experimental group), rhythmical bouts of fictive masticatory-like motoneuron activity (cumulative duration 60-130 min) were induced by electrical stimulation of the left cortical "masticatory area" and recorded from the right digastric motoneuron pool. A control group of five animals (non-masticatory) were treated in the same way as the experimental animals with regard to surgical procedures, anesthesia, paralysis, and survival time. To detect the c-Fos-like protein, the animals were perfused, and the brainstems were cryosectioned and processed immunocytochemically. In the experimental group, the number of c-Fos-like immunoreactive neurons increased significantly in several brainstem areas. In rostral and lateral areas, increments occurred bilaterally in the borderzones surrounding the trigeminal motor nucleus (Regio h); the rostrodorsomedial half of the trigeminal main sensory nucleus; subnucleus oralis-gamma of the spinal trigeminal tract; nuclei reticularis parvocellularis pars alpha and nucleus reticularis pontis caudalis (RPc) pars alpha. Further caudally-enhanced labeling occurred bilaterally in nucleus reticularis parvocellularis and nucleus reticularis gigantocellularis (Rgc) including its pars-alpha. Our results provide a detailed anatomical record of neuronal populations that are correlated with the generation of the masticatory motor behavior. PMID- 15887007 TI - The use of motion cues in the haptic sense of circularity. AB - We tested the hypothesis that speed cues are used to haptically identify changes in the curvature of the hand's trajectory. Subjects grasped the handle of a robotically-controlled manipulandum that was moved in the horizontal plane along various elliptical arcs following one of three different speed profiles. In one profile, a circular arc was traced at a constant speed whereas in the other two speed was constant for ellipses whose aspect ratios differed from unity. A two alternative forced choice procedure was used to identify the ellipse that was sensed to be circular in each of the three experimental conditions. In unconstrained movements, speed varies with the radius of curvature. If speed cues are used to identify curvature during passive movements, one would expect that subjects' responses should be biased towards the ellipse traced at a constant speed. The results did not support this hypothesis, indicating that speed cues are not a major contributor in the haptic sensing of shape. PMID- 15887008 TI - Perceptual coupling in rhythmic movement coordination: stable perception leads to stable action. AB - Rhythmic movement coordination exhibits characteristic patterns of stability, specifically that movements at 0 degrees mean relative phase are maximally stable, 180 degrees is stable but less so than 0 degrees, and other coordinations are unstable without training. Recent research has demonstrated a role for perception in creating this pattern; perceptual variability judgments covary with movement variability results. This suggests that the movement results could be due in part to differential perceptual resolution of the target movement coordinations. The current study used a paradigm that enabled simultaneous access to both perception (between-trial) and movement (within-trial) stability measures. A visually specified 0 degrees target mean relative phase enabled participants to produce stable movements when the movements were at a non-0 degrees relationship to the target being tracked. Strong relationships were found between within-trial stability (the traditional movement measure) and between trial stability (the traditional perceptual judgment measure), suggestive of a role for perception in producing coordination stability phenomena. The stabilization was incomplete, however, indicating that visual perception was not the sole determinant of movement stability. Rhythmic movement coordination is intrinsically a perception/action system. PMID- 15887009 TI - The effect of mild and moderate hepatic impairment on the pharmacokinetics of valdecoxib, a selective COX-2 inhibitor. AB - PURPOSE: This study evaluated the effects of varying degrees of hepatic impairment on the pharmacokinetics and safety of valdecoxib following single and multiple dosing. METHODS: This was an open-label, randomised, parallel group study in 12 subjects with mild hepatic impairment (Child-Pugh Class A) and in 13 with moderate hepatic impairment (Child-Pugh Class B) matched for age, weight, sex, and smoking status; there were two control groups of 12 healthy volunteers, one for each study group. All subjects received a single dose of valdecoxib 20 mg on day 1 and valdecoxib 20 mg twice daily on days 4-7, followed by a single morning dose on day 8. Plasma concentrations of free (unbound) and total valdecoxib and its active hydroxylated metabolite (SC-66905) were measured following single and multiple dosing (day 1 and day 8). Additionally, all subjects received a single intravenous dose of lidocaine 60 mg during the pretreatment period to determine plasma concentrations of monoethylglycinexylidide (MEGX) as a marker of hepatic CYP3A4 activity. RESULTS: The mean apparent oral clearance of free valdecoxib in plasma at steady state decreased by 22-25% in those with mild to moderate impairment (corresponding to a 28-33% increase in the AUC of free valdecoxib and a 19-23% decrease in the AUC of total SC-66905). The mean free fraction of valdecoxib in plasma increased by 9 38%, resulting in a mean decrease in apparent oral clearance of total valdecoxib in plasma of 0-15% (corresponding to a 0-17% increase in the AUC of total valdecoxib). Individual AUCs for free valdecoxib and total SC-66905 did not correlate well with AUCs for MEGX, indicating that decreases in intrinsic clearance of valdecoxib in those with hepatic impairment could not solely be explained by decreased CYP3A4 expression in hepatic impairment. CONCLUSIONS: In our small study sample, mild and moderate hepatic impairment appeared to have only a modest effect on valdecoxib and SC-66905 pharmacokinetics. The adjustment of valdecoxib dose or dosing regimen does not appear mandatory in subjects with mild or moderate hepatic impairment, although caution is necessary during treatment of these patients with valdecoxib. PMID- 15887010 TI - Wave-like appearance of diffuse pachymeningeal enhancement associated with intracranial hypotension. AB - Diffuse pachymeningeal enhancement on magnetic resonance (MR) imaging is important in identifying spontaneous and secondary intracranial hypotension (IH) [cerebrospinal fluid (CSF) hypovolemia] in patients with postural headache, because CSF pressure at lumbar puncture is variable. We examined the pachymeningeal enhancement pattern in patients with IH. MR imaging findings of pachymeningeal enhancement were examined before and after treatment in seven consecutive patients with spontaneous IH and one patient with IH after lumbar puncture. Diffuse non-nodular dural enhancement was observed in all patients. Characteristic thick, uninterrupted, enhancement was observed, mainly in the dura of the frontal, temporal, and retroclival regions, and the tentorium. Thin and uninterrupted, or partially interrupted, enhancement was observed, mainly in the parieto-occipital region and cerebellar convexity. Curved linear enhancement was observed along the calvarium of all patients. A wave-like appearance, a clear pattern of dural unevenness parallel to the brain, was detected in the frontal and temporal regions, near the base, in all patients. A wave-like appearance, especially in the frontal and temporal base, may be a characteristic MR imaging indicator of IH. PMID- 15887011 TI - Prevalence of hydrocephalus in 157 patients with vestibular schwannoma. AB - The purpose of this study was to determine the prevalence of hydrocephalus in patients with vestibular schwannoma. A second objective was to investigate possible etiologies for hydrocephalus in this population by attempting to correlate the incidence and severity of hydrocephalus with tumor volume and extent of fourth ventricular compression. The MRI examinations of 157 adult patients with vestibular schwannoma were retrospectively reviewed. Tumor size was quantified, and the presence of accompanying hydrocephalus was assessed, categorized as communicating type or non-communicating type and then rated as mild, moderate or severe (grades 1-3). Next, the degree of fourth ventricular distortion caused by tumor mass effect was evaluated and categorized as mild, moderate or severe (grades 1-3). Spearman's rank correlation coefficient was used to test the relationships between tumor volume and (1) the extent of fourth ventricular effacement and (2) severity of hydrocephalus. Hydrocephalus was present in 28/157 (18%) cases and was categorized as mild in 11/28 (39%), moderate in 15/28 (54%) and severe in 2/28 (7%). Communicating-type hydrocephalus was present in 17/28 (61%) and non-communicating type in 11/28 (39%). There was a positive correlation between the grade of non-communicating hydrocephalus and tumor volume (r=0.38; P<0.001) and between the severity of fourth ventricular compression and extent of hydrocephalus in this group(r=0.43; P<0.001). In patients who were classified as having communicating hydrocephalus, the correlation between tumor volume and the severity of hydrocephalus was poor (r=0.19; P=0.02) as was the correlation between the extent of fourth ventricular distortion and the severity of hydrocephalus (r=0.21; P<0.01). There is a high prevalence of hydrocephalus in patients with vestibular schwannoma. In a minority of cases non-communicating type hydrocephalus is present and the severity of hydrocephalus can be attributed to the affect of tumor volume on fourth ventricular compression. More commonly, however, communicating-type hydrocephalus exists and the correlation between the severity of fourth ventricular compression and extent of hydrocephalus is poor. Therefore, other etiologies for hydrocephalus, such as tumor protein sloughing, are likely relevant. PMID- 15887012 TI - Endovascular management for intracranial ruptured aneurysms in elderly patients: outcome and technical aspects. AB - The definition of an elderly person is debateable; however, age is a recognised negative prognostic factor for outcome after subarachnoid haemmorrhage, and the age cut-off of 60 years is accepted to define a high risk population. The goal of this article is to access the outcome in this precise population of patients that underwent endovascular treatment (EVT) after aneurysm rupture. Forty-two patients (mean age = 70.24) had 40 aneurysms located at the anterior circulation and nine at the posterior circulation. Thirty-seven (87.9%) patients had Fisher III or IV. Forty-six (93.8%) aneurysms were smaller than 15 mm. Twenty-eight (66.7%) patients were in good neurological state on admission (Hunt and Hess I-III) and 14 (33.3%) in poor state. Satisfactory occlusion rate (total occlusion or neck flow) was achieved in 75% of patients. Follow-up was available in 19 (43.18%) out of the 44 aneurysms treated. Aneurysm recanalization was disclosed in three cases. Satisfactory outcome was achieved on: 60.7% of good grades, 21.4% of poor grades, 43.7% of patients with and 57.6% of patents without comorbidites. Fisher grade (P = 0.0346), comorbidities (P = 0.525) and risk factors (P = 0.515) were not associated with clinical outcome. No age cut-off (65,70 and 75) for favourable outcome could be established, P-values were 0.723, 0.741 and 0.738, respectively. Advancing of age was not associated with an increase number of unfavourable outcome (P = 0.125). Poor neurological status on admission was the only variable associated with unfavourable outcome (P = 0.02). Mortality and morbidity rate related to the procedure were 4.8% and 9.5% respectively. Age should not be taken alone for precluding treatment in ruptured aneurysms, EVT can be considered as a first therapeutic option for elderly persons, since an overall favourable outcome could be achieved in most cases, mainly in non-comatose patients. PMID- 15887013 TI - Chaperone-rich cell lysates, immune activation and tumor vaccination. AB - We have utilized a free-solution-isoelectric focusing technique (FS-IEF) to obtain chaperone-rich cell lysates (CRCL) fractions from clarified tumor homogenates. The FS-IEF technique for enriching multiple chaperones from tumor lysate is relatively easy and rapid, yielding sufficient immunogenic material for clinical use. We have shown that tumor-derived CRCL carry antigenic peptides. Dendritic cells (DCs) uptake CRCL and cross-present the chaperoned peptides to T cells. Tumor-derived CRCL induce protective immune responses against a diverse range of murine tumor types in different genetic backgrounds. When compared to purified heat shock protein 70 (HSP70), single antigenic peptide or unfractionated lysate, CRCL have superior ability to activate/mature DCs and are able to induce potent, long lasting and tumor specific T-cell-mediated immunity. While CRCL vaccines were effective as stand-alone therapies, the enhanced immunogenicity arising from CRCL-pulsed DC as a vaccine indicates that CRCL could be the antigen source of choice for DC-based anti-cancer immunotherapies. The nature of CRCL's enhanced immunogenicity may lie in the broader antigenic peptide repertoire as well as the superior immune activation capacity of CRCL. Exongenous CRCL also supply danger signals in the context of apoptotic tumor cells and enhance the immunogenicity of apoptotic tumor cells, leading to tumor-specific T cell dependent long-term immunity. Moreover, CRCL based vaccines can be effectively combined with chemotherapy to treat cancer. Our findings indicate that CRCL have prominent adjuvant effects and are effective sources of tumor antigens for pulsing DCs. Tumor-derived CRCL are promising anti-cancer vaccines that warrant clinical research and development. PMID- 15887014 TI - T cell-mediated graft-versus-leukemia reactions after allogeneic stem cell transplantation. AB - Allogeneic hematopoietic stem cell transplantation represents the only curative approach for many hematological malignancies. During the last years the impact of the conditioning regimen has been re-assessed. With the advent of reduced intensity conditioning the paradigm has changed from cytoreduction executed by high-dose radio-chemotherapy to immunological surveillance of leukemia by donor cells. Distinct subsets of T cells and NK cells contribute to graft-versus leukemia reactions. So far, cytotoxic T lymphocytes are the mainstay of allogeneic immunotherapy. Here, we summarise the current knowledge of T cell mediated graft-versus-leukemia reactions and present results from pre-clinical and clinical studies of T cell-based adoptive immunotherapy. We address the issues of feasibility and specificity of adoptive immunotransfer from a clinical point of view and discuss the prerequisites for successful clinical applications. Finally, the prospects for immunological research that have evolved with the increasing use of reduced-intensity conditioning and allogeneic stem cell transplantation are highlighted. PMID- 15887015 TI - Analysis of NK cells and chemokine receptors in tumor infiltrating CD4 T lymphocytes in human renal carcinomas. AB - Recent data suggest that chemokines and chemokine receptors mediate leukocyte recruitment of all components of the antitumor response. This study aimed to phenotypically characterize the immune lymphocyte infiltrate in human renal cell carcinomas RCCs and at the invasive margin (tumor-host interface) and to define the association of these findings with established prognostic indicators. Tumor infiltrating lymphocytes TILs were obtained from 24 patients with RCC undergoing radical nephrectomy. Peripheral blood cells from 37 patients were also obtained before surgery. Our findings are consistent with the preferential recruitment of CD4+ Th1-polarized effector memory cells that express CXCR3/CCR5. These cells were the main component of TILs and expressed as CXCR3, CCR5, CD45RO, and CD95. Natural killer (NK) cells were found in significantly higher proportions in TILs of RCCs than in peripheral blood lymphocytes (PBLs) or in other tumors studied (colorectal and breast cancers), where these cells were found in small proportions. No differences in nuclear grade or other studied parameters were observed between the TILs and the lymphocytes present at the invasive margin, which showed a similar composition. However, differences were found according to the tumor stage. First, significantly fewer NK cells were observed in PBLs from metastatic patients. Second, a significantly lower proportion of CCR5/CXCR3/CD4+ cells and a higher proportion of CCR4/CD4+ cells were observed in metastatic patients, suggesting that preferential Th1-polarization may gradually change during the progression of renal cancer cells. Finally, the frequency of CD25/CD4+ cells was higher in metastatic patients. Although the sample of patients with metastasis was small, the overall results suggest a change in composition of the TILs that may potentially confer a selective advantage for tumor growth and may account for the suppression of an effective cytotoxic response. PMID- 15887016 TI - Forphenicinol enhances the antitumor effects of cyclophosphamide in a model of squamous cell carcinoma. AB - We examined the interaction between forphenicinol (FPL) and cyclophosphamide (CPA) or ionizing radiation (IR) on the growth of murine squamous cell carcinoma tumors SCCVII. Primary tumors were established in C3H mice by injecting SCCVII tumor cells subcutaneously into the right hind limb. FPL (100 mg/kg for 8 days) and/or CPA (25 mg/kg twice) were administered by intraperitoneal injection. Tumors were irradiated to a total dose of 40 Gy (eight 5-Gy fractions). SCCVII tumor growth was inhibited by FPL (P=0.054), IR (P=0.003) and CPA (P<0.001) compared with control. The combination of FPL and CPA inhibited tumor growth additively compared with either treatment alone in both small- and large-volume tumors. FPL did not significantly enhance the antitumor effects of IR, however, when CPA+FPL were combined with IR, significant tumor growth inhibition was observed compared with FPL alone (P<0.001), CPA alone (P=0.002) and IR alone (P=0.002). Due to its low toxicity profile, FPL may be combined with CPA, IR and other cytotoxic therapies to potentially enhance the therapeutic ratio. PMID- 15887017 TI - Oxaliplatin causes selective atrophy of a subpopulation of dorsal root ganglion neurons without inducing cell loss. AB - Peripheral neuropathy is induced by multiple doses of oxaliplatin and interferes with the clinical utility of the drug in patients with colorectal cancer. In this study, we sought to determine whether cell loss or selective neuronal damage was the basis for the peripheral neuropathy caused by oxaliplatin. Adult female rats were given 1.85 mg/kg oxaliplatin twice per week for 8 weeks. Nerve conduction and L5 dorsal root ganglia (DRG) were studied 1 week after the completion of all treatment. No mortality occurred during oxaliplatin treatment, but the rate of body weight gain was reduced compared to age-matched vehicle-treated controls. Oxaliplatin slowed conduction velocity and delayed conduction times in peripheral sensory nerves, without affecting central or motor nerve conduction. In L5 DRG, total numbers of neurons were unchanged by oxaliplatin, but there were significant reductions in neuronal size distribution, ganglion volume, average cell size and the relative frequency of large cells. In addition, the relative frequency of small DRG cells was increased by oxaliplatin. Oxaliplatin significantly altered the size distribution and average cell body area of the predominantly large parvalbumin-immunoreactive DRG neurons without affecting the frequency of parvalbumin staining. On the contrary, neither the staining frequency nor the size distribution of the predominantly small substance P immunoreactive DRG neurons was changed by oxaliplatin. In conclusion, oxaliplatin causes selective atrophy of a subpopulation of DRG neurons with predominantly large parvalbumin-expressing cells without inducing neuronal loss. Because DRG cell body size and axonal conduction velocity are positively correlated, neuronal atrophy may be the morphological basis for the development of decreased sensory nerve conduction velocity that characterizes oxaliplatin-induced peripheral neuropathy. PMID- 15887018 TI - Dermal exposure to cyclophosphamide in hospitals during preparation, nursing and cleaning activities. AB - OBJECTIVES: To determine levels of potential and actual dermal exposure to cyclophosphamide (CP) during performance of oncology-related tasks in hospitals and to investigate the relationship with potential sources and surface contamination levels of CP. METHODS: Dermal exposure to CP was determined for tasks with potential exposure to CP: preparation of CP, decanting of patients' urine, washing of the patient, removal of bed sheets of treated patients and cleaning of patients' toilets on oncology wards. Exposure was assessed by the collection of nitrile and latex protective medical gloves (potential exposure), washing of hands (actual exposure), from cotton pads attached to (un)covered forearms (potential or actual exposure) and a wipe sample of the forehead (actual exposure). Bulk samples (i.e. application fluids and patients' excreta) and possible contact surfaces were monitored to assess the amount of CP available for dermal exposure. RESULTS: Pharmacy technicians, oncology nurses and cleaning personnel showed actual and potential dermal exposure to CP during performance of their daily duties. Exposure occurred predominantly on the hands and sporadically on the forehead and forearms. Although all nurses used gloves during handling of patients' urine and sometimes during the other nursing tasks, skin underneath gloves was repeatedly contaminated. Results of tests on bulk and surface contamination samples confirmed that patients intravenously treated with CP excrete the unmetabolised drug, which could subsequently lead to dermal exposure of hospital personnel. A clear relationship was found between dermal exposure levels and direct sources of exposure for all tasks, except for handling patients' urine. CONCLUSIONS: We demonstrated for the first time that actual dermal exposure to CP is common among oncology nurses working with patients treated with this anti-neoplastic drug. Pharmacy technicians and cleaning personnel, on the other hand, are potentially exposed to CP, and protection provided by gloves seemed to be sufficient. PMID- 15887019 TI - Effectiveness of a nationwide interdisciplinary preventive programme for latex allergy. AB - OBJECTIVES: The increasing prevalence of latex allergy among healthcare workers is a large socio-economical problem for the society and affected individuals. The objective of this study was to describe and evaluate a nationwide, interdisciplinary campaign by the institution for the German workers' compensation scheme for non-public healthcare providers that targeted the reduction of exposure to powdered high-protein latex gloves. METHODS: The effectiveness of the prevention programme is described and evaluated with a before-and-after design comparing data on compensation claims for latex-related skin and airway diseases of the German statutory compensation scheme for work related diseases in non-public health services. A survey on change in glove use was conducted after the programme. RESULTS: The main feature of the campaign among healthcare workers (budget 340,000 Euro ) was to increase awareness by means of educational components on the aetiology of latex hypersensitivity for healthcare workers and their providers, and professional trainers. The number of reported compensation claims for latex-induced skin diseases increased from a pre measure of 664 in 1996 to 884 during the programme in 1998 and decreased after the programme (post-measure n = 567 in 1999 and n = 204 in 2002). Doctors' assistants in practices and nurses/nurse assistants/midwives filed most claims. Similar decreases were observed for confirmed claims and latex-related respiratory diseases, while two other prevalent occupational diseases in healthcare workers (low-back disorders, infections) increased during this time period. The survey demonstrated a considerable concurrent drop in the use of powdered latex gloves (hospitals 76% vs 37% for unsterile gloves and 86% vs 62% for sterile gloves). CONCLUSIONS: The campaign against latex allergy in connection with concurrent corresponding recommendations, activities of State authorities for worker protection, and regulations, is an example for the successful incorporation of research results into preventive measures that directly affect the frequency of a work-related disease. This effective programme can be regarded as a model for the reduction of other occupational diseases such as bakers' or isocyanate asthma. PMID- 15887020 TI - The effects of exercise-induced muscle damage on maximal intensity intermittent exercise performance. AB - Exercise-induced muscle damage (EIMD) is a common occurrence following activities with a high eccentric component. Alterations to the torque-velocity relationship following EIMD would appear to have serious implications for athletic performance, particularly as they relate to impairment of maximal intensity exercise. However, this has been studied infrequently. The purpose of this study was to assess the effects of EIMD on maximal intermittent sprint performance. Ten male participants (age 22.4 +/- 3.2 years, height 178.6 +/- 5.2 cm, mass 80.6 +/- 10.7 kg) performed 10 x 6 s cycle ergometer sprints, interspersed with 24 s recovery against a load corresponding to 0.10 kp/kg and 10 x 10 m sprints from a standing start, each with 12 s active (walking) recovery. All variables were measured immediately before and at 30 min, 24, 48 and 72 h following a plyometric exercise protocol comprising of 10 x 10 maximal counter movement jumps. Repeated measures ANOVA showed significant changes over time (all P<0.05) for perceived soreness, plasma creatine kinase activity (CK), peak power output (PPO), sprint time and rate of fatigue. Soreness was significantly higher (P<0.01) than baseline values at all time intervals (3.1, 4.9, 5.5 and 3.2 at 30 min, 24, 48 and 72 h, respectively). CK was significantly elevated (P<0.05) at 24 h (239 IU/l) and 48 h (245 IU/l) compared to baseline (151 IU/l). PPO was significantly lower (P<0.05) than baseline (1,054 W) at all time intervals (888, 946, 852 and 895 W, at 30 min, 24, 48 and 72 h, respectively). The rate of fatigue over the ten cycling sprints was reduced compared to baseline, with the greatest reduction of 48% occurring at 48 h (P<0.01). This was largely attributed to the lower PPO in the initial repetitions, resulting in a lower starting point for the rate of fatigue. Values returned to normal at 72 h. Sprint times over 10 m were higher (P<0.05) at 30 min, 24 h and 48 h compared to baseline (1.96 s) with values corresponding to 2.01, 2.02 and 2.01 at 30 min, 24 h and 48 h, respectively. Values returned to baseline by 72 h. The results provide further evidence that, following a plyometric, muscle-damaging exercise protocol, the ability of the muscle to generate power is reduced for at least 3 days. This is also manifested by a small, but statistically significant reduction in very short-term (approximately 2 s) intermittent sprint running performance. These findings have implications for appropriate training strategies in multiple sprint sports. PMID- 15887021 TI - Changes in force, surface and motor unit EMG during post-exercise development of low frequency fatigue in vastus lateralis muscle. AB - We investigated the effects of low frequency fatigue (LFF) on post-exercise changes in rectified surface EMG (rsEMG) and single motor unit EMG (smuEMG) in vastus lateralis muscle (n = 9). On two experimental days the knee extensors were fatigued with a 60-s-isometric contraction (exercise) at 50% maximal force capacity (MFC). On the first day post-exercise (15 s, 3, 9, 15, 21 and 27 min) rsEMG and electrically-induced (surface stimulation) forces were investigated. SmuEMG was obtained on day two. During short ramp and hold (5 s) contractions at 50% MFC, motor unit discharges of the same units were followed over time. Post exercise MFC and tetanic force (100 Hz stimulation) recovered to about 90% of the pre-exercise values, but recovery with 20 Hz stimulation was less complete: the 20-100 Hz force ratio (mean +/- SD) decreased from 0.65+/-0.06 (pre-exercise) to 0.56+/-0.04 at 27 min post-exercise (P<0.05), indicative of LFF. At 50% MFC, pre exercise rsEMG (% pre-exercise maximum) and motor unit discharge rate were 51.1 +/- 12.7% and 14.1 +/- 3.7 (pulses per second; pps) respectively, 15 s post exercise the respective values were 61.4 +/- 15.4% (P<0.05) and 13.2 +/- 5.6 pps (P>0.05). Thereafter, rsEMG (at 50% MFC) remained stable but motor unit discharge rate significantly increased to 17.7 +/- 3.9 pps 27 min post-exercise. The recruitment threshold decreased (P<0.05) from 27.7 +/- 6.6% MFC before exercise to 25.2 +/- 6.7% 27 min post-exercise. The increase in discharge rate was significantly greater than could be expected from the decrease in recruitment threshold. Thus, post-exercise LFF was compensated by increased motor unit discharge rates which could only partly be accounted for by the small decrease in motor unit recruitment threshold. PMID- 15887022 TI - The reproducibility of an endurance performance test in adolescent cyclists. AB - The purpose of the study was to measure the reproducibility of a performance test in well-trained adolescent cyclists. Eight male and one female cyclist [mean age 15.7 (0.7) y] participated in the study. Lactate threshold (LT) and peak VO2 were assessed. The performance test was repeated on three separate days and consisted of 30 min of steady state (SS) cycling at 80% of individual LT. Immediately after the SS cycling a time trial (TT) started with the cyclists having to complete a fixed amount of work as fast as possible. Reliability was assessed for the TT with the coefficient of variation (CV) as the (SD/mean)*100 for each participant, intraclass correlation coefficients (ICC) and typical error (SD of the difference in mean /radical2). The group mean (SD) times for the TT were TT1 1889 (306), TT2 1857 (283) and TT3 1953 (279) s respectively. Individual CV varied from 0.25% to 10%. The ICC for TT1/2 and 2/3 were r = 0.78 and 0.93 (P<0.05). The typical errors, expressed as a CV% on the log transformed performance times, were 7.3 and 3.7% for TT1/2 and TT2/3 respectively. The largest individual CVs were observed between TT1 and TT2. The differences in CV and SD among the three TTs indicate that trial two and three were more reliable than TT1, suggesting a habituation trial is needed. It is concluded that the present performance test is reliable in adolescent cyclists with lower variation between trials 2 and 3. PMID- 15887024 TI - Supplemental ascorbate and exercise-induced IL-6 metabolism: focus on Fenton chemistry and redox-regulation of vascular homeostasis. PMID- 15887025 TI - Energy cost of swimming of elite long-distance swimmers. AB - The aim of this study was: (1) to assess the energy cost of swimming (C(s), kJ km(-1)) in a group of male (n = 5) and female (n = 5) elite swimmers specialised in long-distance competitions; (2) to evaluate the possible effect of a 2-km trial on the absolute value of C(s). C(s) was assessed during three consecutive 400-m trials covered in a 50-m pool at increasing speeds (v1, v2, v3). After these experiments the subjects swam a 2-km trial at the 10-km race speed (v2km) after which the three 400-m trials were repeated at the same speed as before (v5 = v1, v6 = v2, v7 = v3). C(s) was calculated by dividing the net oxygen uptake at steady state VO2ss by the corresponding average speed (v, m s(-1)). VO2ss was estimated by using back extrapolation technique from breath-to-breath VO2 recorded during the first 30 s of recovery after each test. C(s) increased (from 0.69 kJ m(-1) to 1.27 kJ m(-1)) as a function of v (from 1.29 m s(-1) to 1.50 m s(-1)), its values being comparable to those measured in elite short distance swimmers at similar speeds. In both groups of subjects the speed maintained during the 2-km trial (v2km) was on the average only 1.2% faster than of v2 and v6 (P>0.05), whereas C(s) assessed at the end of the 2-km trial (v2km) turned out to be 21 +/- 26% larger than that assessed at v2 and v6 (P<0.05); the average stroke frequency (SF, cycles min(-1)) during the 2-km trial turned to be about 6% (P<0.05) faster than that assessed at v2 and v6. At v5, C(s) turned out to be 19 +/- 9% (P<0.05) and 22 +/- 27% (0.1 < P = 0.05) larger than at v1 in male and female subjects (respectively). SF was significantly faster (P<0.05, in male subjects) and the distance per stroke (Ds = v/SF) significantly shorter (P<0.05) in female subjects at v5 and v6 than at v1 and v2. These data suggest that the increase of C(s) found after the 2-km trial was likely related to a decrease in propelling efficiency, since the latter is related to the distance per stroke. PMID- 15887026 TI - Distribution of pectic epitopes in cell walls of the sugar beet root. AB - Immunolabelling techniques with antibodies specific to partially methyl esterified homogalacturonan (JIM5: unesterified residues flanked by methylesterified residues. JIM7: methyl-esterified residues flanked by unesterified residues), a blockwise de-esterified homogalacturonan (2F4), 1,4 galactan (LM5) and 1,5-arabinan (LM6) were used to map the distribution of pectin motifs in cell walls of sugar beet root (Beta vulgaris). PME and alkali treatments of sections were used in conjunction with JIM5-7 and 2F4. The JIM7 epitope was abundant and equally distributed in all cells. In storage parenchyma, the JIM5 epitope was restricted to some cell junctions and the lining of intercellular spaces while in vascular tissues it occurred at cell junctions in some phloem walls and in xylem derivatives. After secondary wall formation, the JIM5 epitope was restricted to inner cell wall regions between secondary thickenings. The 2F4 epitope was not detected without de-esterification treatment. PME treatments prior to the use of 2F4 indicated that HG at cell corners was not acetylated. The LM5 epitope was mainly present in the cambial zone and when present in storage parenchyma, it was restricted to the wall region closest to the plasma membrane. The LM6 epitope was widely distributed throughout primary walls but was more abundant in bundles than in medullar ray tissue and storage parenchyma. These data show that the occurrence of oligosaccharide motifs of pectic polysaccharides are spatially regulated in sugar beet root cell walls and that the spatial patterns vary between cell types suggesting that structural variants of pectic polymers are involved in the modulation of cell wall properties. PMID- 15887027 TI - Isolated tumor cells in the bone marrow (ITC-BM) of breast cancer patients before and after anthracyclin based therapy: influenced by the HER2- and Topoisomerase IIalpha-status of the primary tumor? AB - PURPOSE: The presence of isolated tumor cells in the bone marrow (ITC-BM) is an independent prognostic factor in all stages of breast cancer. Both the expression/amplification of human epithelial growth factor receptor 2 (HER2) and Topoisomerase IIalpha (TOP IIa), a key enzyme of DNA replication and main target of anthracyclins, in breast cancer tissue seem to have predictive value regarding the effectiveness of systemic therapies. METHODS: To investigate the correlation between these factors and their influence on clinical outcome, tumor tissue of 54 patients who were screened for ITC-BM before and after anthracyclin-based chemotherapy (abCTX) was examined for HER2 and TOP IIa by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). RESULTS: By IHC, 31% of the tumors showed positive for HER2 (2+/3+), 14.6% were amplified in FISH. TOP IIa expression (>50%) was found in 13/53 patients (25%), FISH was positive in 5/47 cases (11%). TOP IIa amplification was not seen in cases without HER2 amplification, five of the seven HER2 amplified cases also were amplified for TOP IIa (71% co-amplification). Forty-three patients had adjuvant, seven neo adjuvant, four palliative abCTX. ITC-BM were present in 24% of patients before and 31% after CTX. Patients with HER2 (IHC, P = 0.29) and TOP IIa (FISH, P = 0.16) positive tumors tended to stay or become negative in BM status after abCTX and vice versa. After a median follow-up of 44 months (6-127), none of the factors reached significance for overall survival. Yet, patients with HER2 (P = 0.16) and TOP IIa (P = 0.09) positive tumors showed a trend towards prolonged disease-free survival. Remarkably, none of the TOP IIa FISH-positive patients developed distant metastases (P = 0.099) or died (P = 0.19) after CTX so far. CONCLUSIONS: HER2- and TOP IIa positivity seem to improve the effect of abCTX. The combination of the prognostic value of ITC-BM and the predictive capacity of HER2 and TOP IIa could help to stratify patients for certain therapies. The direct examination of those factors on ITC-BM is the focus of ongoing studies. PMID- 15887028 TI - 70 Gy or more: which dose for which prostate cancer? AB - INTRODUCTION: Radical prostatectomy and radiotherapy are currently accepted treatment modalities for localized prostate cancer. Regarding radiotherapy, current evidence suggests that favorable treatment outcome critically depends on adequate radiation doses. However, the exact role of dose in relation to the individual risk profile is complex. In order to evaluate available data on radiation dose response relationships, in prostate cancer, a thorough and critical literature analysis was performed. MATERIAL AND METHODS: Studies on dose response relationships from randomized trials, dose escalation trials, retrospective subgroup analyses and pooled data were identified by Pubmed and ISI web of sciences searches and were critically reviewed. RESULTS AND CONCLUSION: All available data suggest a clear dose response relationship for radiotherapy for localized prostate cancer. In low risk cases, most studies suggest that doses of 70-72 Gy are adequate. Dose escalations up to 78-80 Gy seem to be beneficial for intermediate risk patients. Due to confounding variables, the dose response curves for high-risk patients are less steep. The integration of dose escalation into a more comprehensive treatment protocol is difficult, since trials on the relative impact of either hormonal ablation or inclusion of adjuvant nodal regions on dose escalation are missing. PMID- 15887029 TI - Experimental radioimmunoguided surgery for peritoneal metastases of gastric cancer using anticarcinoembryonic antigen-specific T84.66 F(ab')2. AB - PURPOSE: In gastric cancer, peritoneal dissemination is the most frequent cause of the noncurative resection and recurrence after curative resection. We therefore evaluated the feasibility of radioimmunoguided surgery (RIGS) in the treatment of peritoneal metastases of gastric cancer and the use of anti-CEA specific T84.66 F(ab')2 as an efficient immune agent. METHODS: Two human gastric cancer cell lines, MKN45 and RF48, were intraperitoneally xenografted into nude mice, which were later injected with 125I-labeled T84.66 F(ab')2. Peritoneal tumors were localized by RIGS 5 days after antibody injection. The minimum number of cells detected by a gamma probe was assayed by in vitro tumor cell localization. RESULTS: We observed 37 peritoneal metastases: 8 invisible (long diameter, <1 mm), 6 small (1- < 5 mm), and 23 large (> or =5 mm) tumors. The accuracy, sensitivity and specificity of RIGS in detecting peritoneal metastasis were 82% (69/84), 76% (28/37), and 87% (41/47), respectively. RIGS accuracy did not differ with respect to tumor diameter. Mean labeling indices over minimal and maximal normal counts were 6.1+/-1.2 (mean +/- SEM) and 4.7+/-1, respectively. Mean scores of CEA immunostaining and silver grains in tumors were significantly higher than those in the nontumor-bearing peritoneum (P < 0.001). There was a close correlation among radioactivity, immunostaining and microautoradiography (P < 0.001-0.005). We observed six false-positive and nine false-negatives which may have been due to high blood background and negative radioimmune reactivity, respectively. CONCLUSIONS: 125I-labeled T84.66 F(ab')2 efficiently targeted peritoneally disseminated gastric cancer cells, suggesting that RIGS using this immune agent may accurately detect occult peritoneal metastases in patients with gastric cancer. PMID- 15887030 TI - Degradation of Saccharomyces cervisiae Rck2 upon exposure of cells to high levels of zinc is dependent on Pep4. AB - In undisturbed cells, the MAPK-activated protein kinase Rck2 of Saccharomyces cerevisiae is a stable protein with a turnover time exceeding 60 min. However, we have found that Rck2 is subject to intracellular degradation after exposure of cells to Zn2+ concentrations of 5 mM or more. In high-zinc medium, most of the Rck2 pool is degraded within 5 min. This degradation is blocked by inhibiting the vacuolar proteolytic pathway with the protease inhibitor phenyl methyl sulphonyl fluoride or by mutation of the PEP4 gene. By contrast, blocking the proteasomal pathway with the inhibitor MG132 does not prevent Rck2 degradation upon addition of Zn2+, nor is degradation inhibited in the proteasomal mutations pre1 pre2, cim3, or cim5. The stability of Rck2 is not affected by any of the other stress conditions examined, or by growth rate. Possible mechanisms of the degradation of Rck2 under high zinc conditions, and its physiological significance, are discussed. PMID- 15887031 TI - Comparative phylogenetic analysis of cystatin gene families from arabidopsis, rice and barley. AB - The plant cystatins or phytocystatins comprise a family of specific inhibitors of cysteine proteinases. Such inhibitors are thought to be involved in the regulation of several endogenous processes and in defence against pests and pathogens. Extensive searches in the complete rice and Arabidopsis genomes and in barley EST collections have allowed us to predict the presence of twelve different cystatin genes in rice, seven in Arabidopsis, and at least seven in barley. Structural comparisons based on alignments of all the protein sequences using the CLUSTALW program and searches for conserved motifs using the MEME program have revealed broad conservation of the main motifs characteristic of the plant cystatins. Phylogenetic analyses based on their deduced amino acid sequences have allowed us to identify groups of orthologous cystatins, and to establish homologies and define examples of gene duplications mainly among the rice and barley cystatin genes. Moreover, the absence of a counterpart between the two monocots, as well as strong variations in the motifs that interact with the cysteine proteinases, may be related to a species-specific evolutionary process. This cystatin classification should facilitate the assignment of proteinase specificities and functions to other cystatins as new information is obtained. PMID- 15887032 TI - Functional analysis of the GAF domain of NifA in Azospirillum brasilense: effects of Tyr-->Phe mutations on NifA and its interaction with GlnB. AB - Regulation of NifA activity in Azospirillum brasilense depends on GlnB (a PII protein), and it was previously reported that the target of GlnB activity is the N-terminal domain of NifA. Furthermore, mutation of the Tyr residue at position 18 in the N-terminal domain resulted in a NifA protein that did not require GlnB for activity under nitrogen fixation conditions. We report here that a NifA double mutant in which the Tyr residues at positions 18 and 53 of NifA N-were simultaneously replaced by Phe (NifA-Y1853F) displays high nitrogenase activity, which is still regulatable by ammonia, but not by GlnB. The yeast two-hybrid technique was used to investigate whether GlnB can physically interact with wild type and mutant NifA proteins. GlnB was found to interact directly with the N terminal GAF domain of wild-type NifA, but not with its central or C-terminal domain. GlnB could still bind to the single NifA mutants Y18F and Y53F. In contrast, no interaction was detected between GlnB and the double mutant NifA Y18/53F or between GlnB and NifA-Y43. PMID- 15887033 TI - Differential gene expression during teliospore germination in Ustilago maydis. AB - Ustilago maydis is a model fungal pathogen that induces the formation of tumors in maize. The tumor provides an environment for hyphal differentiation, leading to the formation of thick-walled, diploid teliospores. Such spores serve as a dispersal agent for smut and rust fungi, and their germination leads to new rounds of infection. The morphological changes that occur during teliospore germination in U. maydis have been described in detail. However, the specific molecular events that facilitate this process have not been identified. Through the construction and hybridization of microarrays containing a set of 3918 non redundant cDNAs, we have identified genes that are differentially regulated during teliospore germination. Teliospores induced to germinate for 4 and 11 h were selected for comparison with dormant teliospores. Genes identified as differentially expressed included many that are presumably involved in as yet undescribed molecular events during teliospore germination, as well as characterized genes previously shown to be required for the process. This study represents the first large-scale investigation of changes in gene expression during teliospore germination. PMID- 15887034 TI - A cerebral palsy assessment tool using anatomically based geometries and free form deformation. AB - A geometrical analysis tool for investigating muscle length change in cerebral palsy (CP) patients is presented. A subset of anatomically based geometries from the International Union of Physiological Sciences (IUPS) Physiome Project is used, which is derived from the visible human (VH) data set with muscle attachment information, and customised using volume-preserving free-form deformation (FFD), the 'host-mesh' technique. The model's intended use is to provide pre- and post-surgery assessment for muscle lengthening, a surgery performed to help slacken tight muscles and improve gait. The model is illustrated using healthy patient data from motion capture as a validation followed by three CP case studies to highlight its use. The methodology is presented in three stages, (1) a FFD of the complete lower limb, (2) a focused geometric study on the semimembranosus (SM) and gastrocnemius (GT) muscles, and (3) an improved hybrid mechanics-FFD approach as an improvement for future analysis, with differentiation between muscle and tendon lengthening, and contact detection between sliding muscles. Finally, the issues, limitations, in particular with the marker system, and model improvements are discussed. PMID- 15887035 TI - Impact of carbon and nitrogen nutrition on the quality, yield and composition of blastospores of the bioinsecticidal fungus Paecilomyces fumosoroseus. AB - The impact of growing cultures of Paecilomyces fumosoroseus in liquid media containing four combinations of glucose and casamino acids (8 g l(-1) or 80 g l( 1) glucose, 1.32 g l(-1) or 13.2 g l(-1) casamino acids) was evaluated, based on blastospore production, germination rate, viability after freeze-drying and short term storage stability. When blastospores were produced using a high casamino acid concentration, blastospore yields and germination rates were significantly higher (13.2-18.5 x 10(7) blastospores ml(-1), 50-60% germination after 4 h), compared to cultures grown in media containing lower casamino acid concentrations (0.4-2.3 x 10(7) blastospores ml(-1), 10-20% germination after 4 h). Chemical analyses of blastospore composition showed that accelerated blastospore germination may be related to increased proteinaceous reserves rather than to glycogen or lipid accumulation. Tolerance to freeze-drying by blastospores suspended in spent medium was enhanced by a high initial casamino acid concentration in the culture medium (75% survival) and by the residual glucose concentrations in the spent medium. Under the conditions of this study, the storage stability of blastospores of P. fumosoroseus was unaffected by the nutritional condition in which they were produced. PMID- 15887036 TI - Wavelet-based noise reduction for improved deconvolution of time-series data in dynamic susceptibility-contrast MRI. AB - Dynamic susceptibility-contrast (DSC) MRI requires deconvolution to retrieve the tissue residue function R(t) and the cerebral blood flow (CBF). In this study, deconvolution of time-series data was performed by wavelet-transform-based denoising combined with the Fourier transform (FT). Traditional FT-based deconvolution of noisy data requires frequency-domain filtering, often leading to excessive smoothing of the recovered signal. In the present approach, only a low degree of regularisation was employed while the major noise reduction was accomplished by wavelet transformation of data and Wiener-like filtering in the wavelet space. After inverse wavelet transform, the estimate of CBF.R(t) was obtained. DSC-MRI signal-versus-time curves (signal-to-noise ratios 40 and 100) were simulated, corresponding to CBF values in the range 10-60 ml/(min 100 g). Three shapes of the tissue residue function were investigated. The technique was also applied to six volunteers. Simulations showed CBF estimates with acceptable accuracy and precision, as well as independence of any time shift between the arterial input function and the tissue concentration curve. The grey-matter to white-matter CBF ratio in volunteers was 2.4+/-0.2. The proposed wavelet/FT deconvolution is robust and can be implemented into existing perfusion software. CBF maps from healthy volunteers showed high quality. PMID- 15887037 TI - RGA- and RAPD-derived SCAR markers for a Brassica B-genome introgression conferring resistance to blackleg in oilseed rape. AB - An introgression derived from the B genome of Brassica juncea in spring-type oilseed rape (B. napus) conferring recessively inherited cotyledon resistance against several pathotypes of the blackleg fungus Leptosphaeria maculans was mapped using PCR-based molecular markers. Resistance-associated B-genome-specific randomly amplified (RAPD) and resistance gene analog (RGA) DNA polymorphisms were converted into three sequence-specific markers (SCARs; B5-1520, C5-1000, RGALm). The flanking sequence of the RGALm locus was determined by genomic walking, leading to a 1,610-bp EcoRV fragment which showed extensive homology to known and putative resistance genes of a cluster on Arabidopsis chromosome 5. Partial sequence analysis of the genomic RAPD segment OPC-05-1700 revealed strong homology to the gibberellin 2-oxidase gene of Arabidopsis. The SCAR markers were analyzed in two segregating populations and were found to be linked in coupling to each other, and in repulsion to the resistance locus. In both populations, markers deviated significantly from a monogenic 3:1 segregation ratio, with plants lacking the markers being more frequent than expected. Although the mode of introgression is yet unknown, the recombinant individuals observed among susceptible progeny suggest homeology between the B-genome-specific segment and its B. napus counterpart. This would offer prospects for reducing the size of the introgression and further fine mapping of the resistance locus. PMID- 15887038 TI - SSR allelic diversity changes in 480 European bread wheat varieties released from 1840 to 2000. AB - A sample of 480 bread wheat varieties originating from 15 European geographical areas and released from 1840 to 2000 were analysed with a set of 39 microsatellite markers. The total number of alleles ranged from 4 to 40, with an average of 16.4 alleles per locus. When seven successive periods of release were considered, the total number of alleles was quite stable until the 1960s, from which time it regularly decreased. Clustering analysis on Nei's distance matrix between these seven temporal groups showed a clear separation between groups of varieties registered before and after 1970. Analysis of qualitative variation over time in allelic composition of the accessions indicated that, on average, the more recent the European varieties, the more similar they were to each other. However, European accessions appear to be more differentiated as a function of their geographical origin than of their registration period. On average, western European countries (France, The Netherlands, Great Britain, Belgium) displayed a lower number of alleles than southeastern European countries (former Yugoslavia, Greece, Bulgaria, Romania, Hungary) and than the Mediterranean area (Italy, Spain and Portugal), which had a higher number. A hierarchical tree on Nei's distance matrix between the 15 geographical groups of accessions exhibited clear opposition between the geographical areas north and south of the arc formed by the Alps and the Carpathian mountains. These results suggest that diversity in European wheat accessions is not randomly distributed but can be explained both by temporal and geographical variation trends linked to breeding practices and agriculture policies in different countries. PMID- 15887039 TI - Designing a microarray experiment to estimate dominance in maize (Zea mays L.). AB - Experiments using cDNA microarrays for the identification of genes with certain expression patterns require a thoughtfully planned design. This study was conducted to determine an optimal design for a microarray experiment to estimate differential gene expression between hybrids and their parental inbred lines in maize (i.e. dominance). It has two features: the contrasts of interest contain more than two genotypes and the procedure may be customised to other microarray experiments where different effects may influence hybridisation signals. A mixed model was used to include all important effects. Impacts during growth of the plant material were taken into consideration as well as those occurring during hybridisation. The results of a preliminary experiment were used to determine which effects were to be included in the model, and data from another microarray experiment were used to estimate variance components. In order to select good designs, an optimality criterion adapted to the problem of differential gene expression between hybrids and their parental inbred lines was defined. Two approaches were used to determine an optimal design: the first one simplifies the problem by dividing it into several subproblems, whereas the second is more sophisticated and uses a simulated annealing (SA) algorithm. We found that the first approach constitutes a useful means for designing microarray experiments to study this problem. Using the more sophisticated SA approach the design can be further improved. PMID- 15887040 TI - Genetic relationships between resistance to stalk-tunneling by the European corn borer and cell-wall components in maize population B73xB52. AB - The objective of this study was to assess the relationships among quantitative trait loci (QTL) detected for European corn borer (ECB) tunneling and cell-wall components (CWC) neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL) content in leaf-sheath and stalk tissues in a maize recombinant inbred line population derived from inbred lines B73 and B52. Most of the QTL for ECB resistance (10/13) were at QTL positions for one or more CWC. Of the 12 QTL for NDF and ADF in leaf-sheaths, five for each trait were at or near QTL for ECB tunneling. Four of these five QTL for NDF and ADF mapped to common locations. Four of the eight leaf-sheath ADL QTL were detected in the same genomic regions as ECB QTL. For stalk tissue, four regions contained common/overlapping QTL for ECB tunneling, NDF, and ADF. Six such regions were observed for stalk ADL and ECB tunneling. Seven of the ten QTL associated with both CWC and ECB tunneling contributed to the negative correlations observed between these traits, while relatively few QTL effects were positively correlated. This suggests that while CWC contribute to ECB resistance in this population, other mechanisms and other genes also are involved. Several QTL contributing to the negative correlations between ECB tunneling and CWC in the leaf-sheaths mapped to similar positions as QTL detected in tropical maize populations for resistance to leaf-feeding by Diatraea grandiosella Dyar and Diatraea saccharalis Fabricus. These regions may contain genes involved in the synthesis of cellulose, hemicellulose, and lignin in the leaf-blades and leaf sheaths of maize. PMID- 15887041 TI - Variant MoFe proteins of Azotobacter vinelandii: effects of carbon monoxide on electron paramagnetic resonance spectra generated during enzyme turnover. AB - The resting state of wild-type nitrogenase MoFe protein exhibits an S=3/2 electron paramagnetic resonance (EPR) signal originating from the FeMo cofactor, the enzyme's active site. When nitrogenase turns over under CO, this signal disappears and one (sometimes two) of three new EPR signals, which also arise from the FeMo cofactor, appears, depending on the CO concentration. The appearance and properties of these CO-inducible EPR signals, which were also generated with variant MoFe proteins (alphaR96Q, alphaR96K, alphaQ191K, alphaR359K, alphaR96K/alphaR359K, alphaR277C, alphaR277H, and DeltanifV) that are impacted around the FeMo cofactor, have been investigated. No new CO-induced EPR signals arise from any variant, suggesting that no new CO-binding sites are produced by the substitutions. All variant proteins, except alphaR277H, produce the lo-CO signal; all, except alphaQ191K, produce the hi(5)-CO signal; but only two (alphaR96Q and DeltanifV) exhibit the hi-CO signal. FeMo cofactor's environment clearly dictates which CO-induced EPR signals are generated; however, none of these EPR signals correlate with CO inhibition of H(2) evolution observed with some of these variants. CO inhibition of H(2) evolution is, therefore, due to CO binding to a different site(s) from those responsible for the CO-induced EPR signals. Some resting-state variants have overlapping S=3/2 EPR signals, whose intensities simultaneously decrease under turnover conditions, indicating that all FeMo cofactor conformations are catalytically active. Moreover, these variants produce a similar number of hi(5)-CO signals after turnover under CO to the number of resting-state S=3/2 signals. The FeMo cofactor associated with the hi(5)-CO signal likely contains two bridging CO molecules. PMID- 15887042 TI - Angiotensin-cleaving catalysts: conversion of N-terminal aspartate to pyruvate through oxidative decarboxylation catalyzed by Co(III)cyclen. AB - To provide a firm basis for the new paradigm of drug discovery based on peptide cleaving catalysts, oligopeptide-cleaving catalysts were searched for by using human angiotensin I (Ang-I) and angiotensin II (Ang-II) as the substrates. Catalyst candidates containing the Co(III) complex of cyclen as the catalytic center were prepared by multicomponent condensation reactions. From two types of chemical libraries containing about 3,600 catalyst candidates, two compounds [SS Co(III)X and S-Co(III)Y] were selected as the most active catalysts. On incubation with SS-Co(III)X and S-Co(III)Y, both Ang-I and Ang-II were cleaved by oxidative decarboxylation instead of peptide hydrolysis: the N-terminal Asp residues of Ang-I and Ang-II were converted to pyruvate residues. Catalysts for oxidative decarboxylation of the N-terminal Asp residue contained in an oligopeptide are unprecedented in both biological and chemical systems. Detailed kinetics analysis suggested that Ang-I and Ang-II can be cleaved with half-lives much less than 1 h if the structures of the chelating ligands of the catalysts are further improved. The results indicated that the concept of the peptide cleaving catalysts can be expanded to include oligopeptides as the targets and nonhydrolytic reactions as the means for cleavage. PMID- 15887043 TI - Rosiglitazone up-regulates lipoprotein lipase, hormone-sensitive lipase and uncoupling protein-1, and down-regulates insulin-induced fatty acid synthase gene expression in brown adipocytes of Wistar rats. AB - AIMS/HYPOTHESIS: Although thiazolidinediones are now widely used to treat type 2 diabetes, their mechanism of action remains largely unknown. They are agonists for the transcription factor PPARgamma, and in addition to their insulin sensitising effects, they can promote adipogenesis and control gene expression in adipose tissues. We have explored the effect of rosiglitazone on insulin-mediated induction of pivotal genes involved in lipid metabolism and thermogenesis in brown fat. The genes studied were: (1) lipoprotein lipase (lpl), which is involved in lipid uptake; (2) hormone-sensitive lipase (hsl), which mobilises fatty acids from stored triglycerides; (3) fatty acid synthase (fas), which regulates de novo lipogenesis; and (4) the uncoupling proteins (ucp) 1 and 3, which control thermogenesis. METHODS: We used fetal rat primary brown adipocytes cultured with insulin, rosiglitazone or both combined. Then, we studied gene expression by northern and western blotting, as well as 'run-on' and gel-shift assays to identify binding of potential transcription factors to the fas promoter. RESULTS: Exposure to rosiglitazone for 24 h induced ucp-1, lpl and hsl gene expression and when rosiglitazone was combined with insulin a synergistic effect on lpl and ucp-3 mRNA expression was produced. These effects were consistent with increased LPL and HSL activities as well as respiration rates, mainly in response to exogenous palmitate. In contrast, treatment with rosiglitazone did not alter FAS mRNA basal levels but prevented the induction elicited by insulin in a time- and dose-dependent manner. Correspondingly diminished FAS protein levels and activity, as well as cellular lipid content, were observed, indicating an antilipogenic action of rosiglitazone in brown adipocytes. Furthermore, rosiglitazone impaired insulin increase in the FAS transcription rate by antagonising insulin-induced binding of upstream stimulatory factors to the E-box consensus sequence in the FAS promoter and insulin-induced binding of activating protein-1. CONCLUSIONS/INTERPRETATION: Rosiglitazone prevents insulin-induced up-regulation of the main lipogenic enzyme but increases the expression of those enzymes involved in lipid uptake and mobilisation, favouring fatty acid utilisation through uncoupled respiration. PMID- 15887044 TI - Correlation of different bone markers with bone density in patients with rheumatic diseases on glucocorticoid therapy. AB - Osteoporosis is a common concomitant disease in patients with rheumatic diseases on glucocorticoid (GC) therapy. Bone status is usually evaluated by determination of bone density in combination with clinical examinations and laboratory tests. However, the strength of individual biochemical bone makers in GC-induced osteoporosis has yet to be fully clarified. For this reason, different bone markers were investigated in correlation with bone density in patients with rheumatic diseases. Approximately 238 patients (212 women, 26 men) with a rheumatic disease and under GC therapy were examined consecutively for the first time with regard to bone density (BMD) and bone markers [osteocalcin, bone specific alkaline phosphatase (precipitation method/tandem-MP ostase), crosslinks [pyridinoline (PYD), deoxypyridinoline (DPX), N-terminal telopeptide (NTX)]]. The daily glucocorticoid dose was 10 mg prednisone equivalent (median), and the cumulative dose was 12 g prednisone equivalent (median). None of the patients had previously taken medication for osteoporosis. Osteoporosis was demonstrated in 35.3% of the patients, osteopenia in 47.5%, and a normal BMD in 17.2%. The results of tandem-MP ostase correlated with the BMD of the lumbar spine and of the femoral neck. The values for N-terminal telopeptide and pyridinoline correlated only with the bone density of the femoral neck. All results were statistically significant, although the correlation coefficients were low. After classification of the patients according to their BMD values (osteoporosis, osteopenia and normal BMD), there were significantly more patients with bone markers above the norm in the osteoporosis group and in the osteopenia group than in the group with normal bone density. All bone markers recorded behaved similarly in relation to the bone density values. The same analysis was also undertaken for the different disease groups. In these subgroups there was also a correlation between ostase/crosslinks with BMD, but the correlation coefficients were low. A general recommendation for the routine use of a specific bone marker in patients with rheumatic diseases on glucocorticoid therapy cannot be made from a cost-benefit point of view mainly because of limited predictive power (low correlation coefficients, incomplete correlation with different sites of BMD measurement). PMID- 15887045 TI - Expression and function of Bapx1 during chick limb development. AB - The homeobox-containing transcription factor Bapx1 (also known as Nkx3.2) is crucial for development of the axial skeleton and parts of the chondrocranium. Here we describe the detailed expression of Bapx1 during chick limb development and show that in contrast to its expression in the axial skeleton, Bapx1 is expressed after the commitment to chondrogenesis. Bapx1 is initially expressed throughout the developing skeletal elements prior to the overt differentiation of the distinct chondrogenic layers. Once distinct layers (proliferating, prehypertrophic and hypertrophic) have formed, Bapx1 expression is restricted to the proliferating chondrocytes. Bapx1 transcripts are excluded from the articular cartilage. A second homeobox-containing transcription factor, Barx1, is expressed in a complementary fashion in the developing joint and articular cartilage. Interestingly, in vitro functional analyses showed that Bapx1 overexpression in micromass cultures increased both matrix production and nodule number suggesting that Bapx1 is sufficient to promote chondrogenesis in the limb. In contrast, Barx1 had the opposite effect on nodule number suggesting that it has an inhibitory effect on chondrogenic initiation consistent with its expression in the developing joint. A slight increase in matrix levels was also observed consistent with its expression in the articular chondrocytes. Finally, we show that Bapx1 is also expressed in the soft tissues such as the developing tendons, muscle sheaths and surrounding mesenchyme, and therefore may have additional as yet uncharacterized roles in limb morphogenesis. PMID- 15887046 TI - The autonomic nervous system of the human heart with special reference to its origin, course, and peripheral distribution. AB - A submacroscopic anatomical investigation of the entire autonomic cardiac nervous system, from origin to peripheral distribution, was performed by examining 36 sides of 18 adult human cadavers under a stereomicroscope. The following new results and points of discussion were obtained: (1) The superior cervical, the middle cervical, the vertebral, and the cervicothoracic (stellate) ganglia, composed of the inferior cervical and 1st thoracic ganglia, were mostly consistent among the specimens. (2) The superior, middle, and inferior cardiac nerves innervated the heart by simply following the descent of the great arteries. In contrast, the thoracic cardiac nerve in the posterior mediastinum followed a complex course because of the long distance to the middle mediastinum. (3) The actual course of the right thoracic cardiac nerve differed from that of the previous descriptions in that it ascended obliquely or ran transversely to the vertebrae, regardless of the intercostal vessels. Regarding the right thoracic cardiac nerve, two descending courses were observed: the descent of the right thoracic cardiac nerve via the azygos vein and right venous porta, and the descent of the recurrent right thoracic cardiac nerve via the aorta. (4) The cranial cardiac nerve and branch tended to distribute into the heart medially, and the caudal cardiac nerve and branch tended to distribute into the heart laterally. (5) The mixing positions (cardiac plexus) of the sympathetic cardiac nerve and the vagal cardiac branch, as well as the definitive morphology of brachial arteries with the recurrent laryngeal nerves, tended to differ on both sides. These new and detailed anatomical descriptions of the human autonomic cardiac nervous system may provide important clues regarding the morphogenesis of autonomic cardiac nerves in addition to contributing to the improvement of cardiac surgery. PMID- 15887047 TI - Delayed postnatal settlement of cerebellar Purkinje cells in vermal lobules VI and VII of the mouse. AB - The postnatal development of the ganglionic (Purkinje) layer was studied in the mouse cerebellum from P0 to young adulthood with special emphasis to vermal lobules VI-VII (oculomotor vermis) in the mouse. In order to visualize Purkinje cells (PCs), toluidine blue staining of resin-embedded semithin sections and calbindin immunohistochemistry were utilized. The number of PCs in the whole cerebellum was 199,080+/-2966 at postnatal day eight (P8), 222,000+/-2979 at P20 and nearly the same, 225,800+/-7549 in young adults; i.e., there was an approximately 13.4% increase of PCs between P8 and adults. The number of PC somata aligned into a rostrocaudal stripe along the developing ganglionic layer increased by about 24% in vermal cerebellar lobule III but much more markedly (i.e., by 49%) in VI+VII between P6 and young adulthood. Between P6 and P16, the increase of the number of PCs in the ganglionic layer of lobules VI and VII resulted in the (delayed) completion of PC layer, caused by the (late) alignment of rostrocaudally dispersed PCs, although late postnatal migration of a smaller population of these cells cannot be excluded either. It is concluded that the oculomotor vermis belongs to the latest developing cerebellar cortical structures, which could be the reason for its frequent involvement in developmentally related disturbances and disorders. PMID- 15887048 TI - [The NMDA receptor system: genetic risk factor for alcoholism]. AB - Alcohol dependence is one of the most common addictive diseases and known to be in part genetically transmitted, based on an oligogenic background in which each gene involved contributes only little to the resulting phenotype. Besides influencing other signal transduction mechanisms, alcohol specifically inhibits the NMDA signaling cascade, which mediates the excitatory effects of glutamate in the brain. Target molecules, sensitive to ethanol, include the NMDA receptors as well as downstream molecules of the glutamatergic system, glutamate transporters, and associated regulatory proteins. Adaptive processes of the glutamatergic system during chronic alcohol consumption may play a major role for later development of reward symptoms. Candidate gene studies, including association studies and animal models, are powerful and sensitive for detecting oligogenic effects and thus important to alcoholism research. PMID- 15887049 TI - [Extraordinary cause of a self-induced focal epilepsy]. AB - Epilepsies and psychiatric disorders such as psychoses are known to have reciprocal influence. There are problems in patient compliance and pharmacological interactions between the drugs used for treatment. We report an unusual, self-induced, focal, symptomatic epilepsy in a hitherto unrevealed schizophrenic psychosis. The interdisciplinary treatment of three different departments was necessary. The initial symptom of a diffuse frontal headache and the patient's penetration of his own skull in an attempt at self-treatment led to the development of a frontal abscess around a foreign body and the described focal epilepsy. PMID- 15887050 TI - [Treatment utilisation of a psychoeducational group for relatives of patients with affective disorders]. AB - The relevance of family interactions in the course of affective disorders has been well described. In contrast to the situation regarding schizophrenic disorders, there are few systematic concepts for involvement of the relatives of patients with affective disorders in treatment. The goal of this study was the development and evaluation of a standardised psychoeducational treatment programme. We determined the number and characteristics of relatives accepting the offer of such a group. Relatives of almost half of 55 patients with major depression and a bipolar disorder participated in the group. Relatives of male patients were more likely to take part than relatives of female patients. Relatives of patients with a bipolar disorder were more likely to take part than relatives of patients with unipolar depression. The patients whose relatives attended the group showed a more favourable understanding of the illness and more knowledge about affective disorders, but on the other hand, felt themselves to be more strongly criticised by their relatives and had less social support than the other patients. These results emphasise the importance of differential family focused treatment modalities in affective disorders. PMID- 15887051 TI - [Sentinel node biopsy. What are the facts?]. AB - Sentinel node biopsy (SNB) is increasingly being used as a minimally invasive staging procedure in patients with malignant melanoma. For decades elective lymph node dissection (ELND) was performed in many centers on patients at risk for lymph node metastasis but without clinically detectable lymph node involvement. Today, selective lymph node dissection (SLND) is offered only to patients with histologically proven metastasis in a SN (10-29%). A positive SN is one of the most important prognostic parameters. Ten years after the introduction of the technique, the role of SNB in the treatment of cutaneous melanoma still remains controversial. Issues include the usefulness of highly sensitive evaluation of SN using molecular biology or cytology techniques, as well as the therapeutic impact of the SNB per se and the associated combined surgical or medical adjuvant therapies. PMID- 15887052 TI - [Modern aspects of varicose vein surgery]. AB - In Germany almost every third adult suffers from varicose veins requiring treatment. Conventional varicose vein surgery by high ligation and stripping is widely accepted as standard therapy for saphenous vein insufficiency, although associated with a high frequency of recurrent varicosities. Innovative endovascular procedures laying claim to be minimally invasive have been implemented over the last five years: endovenous radiofrequency obliteration, endovenous laser treatment and ultrasound-guided sclerotherapy with foam. The early treatment outcomes are promising in regard to recurrent varicose veins, cosmetic results and convalescence. Evidence-based prospective trials with large numbers of participants comparing the interventional procedures with high ligation and stripping are still missing. This report delineates current developments in varicose vein surgery and provides information on principles, effectiveness and side effect profiles of endovascular therapy procedures. PMID- 15887053 TI - [Ambulatory dermatologic surgery]. AB - Outpatient surgery in dermatology is likely to become increasingly important in future, for a number of reasons. Current social legislation favors outpatient surgery, because hospitals must justify performing any procedure in the outpatient catalogue on an inpatient basis. The range of conditions being treated is also broadening, for example due to the growing popularity of aesthetic enhancement procedures. Epidemiological trends in skin cancer will also lead to considerable growth in demand for outpatient dermatological surgery. This article analyses current trends in ambulatory dermato-surgery, taking into account social, legislative, hygienic, legal and economic aspects. Measures to promote quality assurance are also described. The difficult cost situation facing outpatient surgery is creating a need for new forms of cooperation between primary care physicians and hospitals. PMID- 15887055 TI - Effects of caffeine and caffeine withdrawal on mood and cognitive performance degraded by sleep restriction. AB - RATIONALE: It has been suggested that caffeine is most likely to benefit mood and performance when alertness is low. OBJECTIVES: To measure the effects of caffeine on psychomotor and cognitive performance, mood, blood pressure and heart rate in sleep-restricted participants. To do this in a group of participants who had also been previously deprived of caffeine for 3 weeks, thereby potentially removing the confounding effects of acute caffeine withdrawal. METHODS: Participants were moderate to moderate-high caffeine consumers who were provided with either decaffeinated tea and/or coffee for 3 weeks (LTW) or regular tea and/or coffee for 3 weeks (overnight caffeine-withdrawn participants, ONW). Then, following overnight caffeine abstinence, they were tested on a battery of tasks assessing mood, cognitive performance, etc. before and after receiving caffeine (1.2 mg/kg) or on another day after receiving placebo. RESULTS: Final analyses were based on 17 long-term caffeine-withdrawn participants (LTW) and 17 ONW participants whose salivary caffeine levels on each test day confirmed probable compliance with the instructions concerning restrictions on consumption of caffeine-containing drinks. Acute caffeine withdrawal (ONW) had a number of negative effects, including impairment of cognitive performance, increased headache, and reduced alertness and clear-headedness. Caffeine (versus placebo) did not significantly improve cognitive performance in LTW participants, although it prevented further deterioration of performance in ONW participants. Caffeine increased tapping speed (but tended to impair hand steadiness), increased blood pressure, and had some effects on mood in both groups. CONCLUSIONS: The findings provide strong support for the withdrawal reversal hypothesis. In particular, cognitive performance was found to be affected adversely by acute caffeine withdrawal and, even in the context of alertness lowered by sleep restriction, cognitive performance was not improved by caffeine in the absence of these withdrawal effects. Different patterns of effects (or lack of effects) of caffeine and caffeine withdrawal were found for other variables, but overall these results also suggest that there is little benefit to be gained from caffeine consumption. PMID- 15887056 TI - Tryptophan depletion reduces right inferior prefrontal activation during response inhibition in fast, event-related fMRI. AB - RATIONALE AND OBJECTIVE: In animal and human studies, the neurotransmitter serotonin (5-hydroxytryptamine; 5-HT) has been implicated in mediating impulsiveness and aggression. To test the hypothesis that 5-HT modulates neuro cognitive brain activation during inhibitory control, we examined the effect of acute tryptophan depletion (ATD), a dietary challenge, which has been shown to decrease 5-HT synthesis in the brain, on functional brain activation during a go/no-go task. METHODS: Nine healthy, right-handed volunteers performed a rapid, event-related go/no-go task in two functional magnetic resonance imaging (fMRI) scanning sessions, 5 h after either a tryptophan-free or a balanced amino acid drink in a double-blind, sham depletion-controlled, counterbalanced, crossover design. The task required subjects to selectively execute or inhibit a motor response. Tryptophan depletion significantly lowered total plasma tryptophan concentration by 80%, but did not significantly alter inhibitory performance or mood ratings. RESULTS: ATD significantly reduced right orbito-inferior prefrontal activation during the no-go condition, and increased activation in superior and medial temporal cortices. CONCLUSIONS: These findings provide neuro-functional evidence of a serotonergic modulation of right inferior prefrontal during inhibitory motor control. The increased engagement of temporal brain regions may reflect compensatory mechanisms. PMID- 15887057 TI - Can a subgroup of OCD patients with motor abnormalities and poor therapeutic response be identified? AB - RATIONALE: In a subgroup of patients with obsessive-compulsive disorder (OCD), motor soft signs, tics and other movement disorders can be observed, indicating a special pathogenetic involvement of basal ganglia. OBJECTIVES: The main objective of this study was to verify the hypothesis that such motor dysfunction characterises a subgroup of OCD patients with poor treatment response. For assessing even subtle motor dysfunction, a new method for kinematical analysis of hand movements has been applied. METHODS: We examined the performance of 45 in patients who met the DSM-IV criteria for OCD before and under therapy (sertraline and behaviour therapy) using a digitising tablet and kinematical analysis of simple handwriting and drawing movements. All subjects wrote a sentence, their signature and letter sequences. Moreover, they drew circles under different conditions. Three kinematical parameters (stroke duration, variation coefficient of peak velocity, stroke length) were calculated to quantify hand-motor performance. RESULTS: Prior to therapy, non-responders wrote with significantly smaller amplitudes than responders. Additionally, non-responders drew significantly larger circles with the non-dominant hand at baseline, as compared to responders. Disturbances of handwriting were more frequent in non-responders than in responders. CONCLUSIONS: Kinematical analysis of handwriting movements seems to be interesting for the prediction of poor response to treatments in OCD patients. PMID- 15887058 TI - [Clinical trials]. PMID- 15887059 TI - [The geriatric patient -- medical entitlements and reality]. PMID- 15887060 TI - [Clinical trials. An overview]. AB - Since the mid-1950s, the basis for evaluating medical therapies has shifted from transferred knowledge, subjective impression and personal experience towards clinical trials, first, in reaction to side effect tragedies, in drug licensing, then to a growing extent in other parts of medicine such as the assessment of medical devices or surgical procedures. Clinical trials are prospective studies of the efficacy and safety of therapies by repeated application. For this purpose, therapies are usually compared to controls, based on statistical reasoning. Initial comparability of study groups is best guaranteed by randomisation while equality of observation and of concomitant treatment is best guaranteed by blinding. Clinical trials do not result in clear-cut yes/no answers, but reduce the range of reasonable interpretations regarding efficacy and safety of a therapy. Thus, they allow decisions to be made on solid objective grounds. In this way, clinical trials have contributed substantially to medical progress over the last 50 years and are indispensable for today's medical research. PMID- 15887061 TI - [Clinical trials for medicinal products. New European and national legal rules]. AB - With the 12th Amendment of the German Drug Law, in addition to other changes, transformation requirements resulting from the Directive 2001/20/EU of the European Parliament and the Council on clinical trials for medicinal products for humans have been fulfilled. Resulting changes in this context for competent federal authorities and other institutions involved are briefly presented and their relevance discussed. These new legal regulations provide a number of opportunities to plan clinical trials more effectively and to perform them in a more structured and better harmonized way, thus leading to increased safety for patients and more reliable results. PMID- 15887062 TI - [Clinical trials for children]. AB - The 12th AMG amendment and the EU Directive 2001/20/EC set down clear legal guidelines on clinical studies of drug products in children. As a consequence there are new prospects for an increasing involvement of the pharmaceutical industry in developing drugs for use in children. BfArM has acted on this development by setting up a committee of experts on drug approval for use in children-similar to committees at the EMEA and FDA working on the same issue. Another useful step consists in the strengthening of pharmaco-vigilance measures. Treatment standards have been developed on the basis of the research instrument "clinical therapy optimization studies", frequently conducted as co-operative national or international studies joining many trial sites, and may definitely benefit from the new requirements. Clearly defined terms and regulations, such as "sponsor responsibility", "trial approval", "investigational medical product", "insurance", and "monitoring" describe problems that can or at times cannot be solved by such research groups. The sponsor's responsibilities in an international trial e. g. cannot usually be shouldered by a single scientist or treatment center. When in the future practically every single research question on drug treatment is governed by those regulations, costs will soar and fewer questions will be asked in clinical studies. So far, the AMG amendment has not given an assessment of benefit and risk regarding treatment quality and drug safety in children. Such an assessment depends on the willingness of all concerned to explore the scope of the legal regulations in order to let children participate in scientific progress where it is needed and where it makes sense. PMID- 15887063 TI - [Gender-related differences in pharmacokinetics and pharmacodynamics]. AB - Following the negative experience with thalidomide, women were excluded from participation in clinical trials with new pharmaceutical agents as far as possible, especially from phase I studies. However, in the early 1990s a body of evidence accumulated suggesting clinically relevant gender-related differences in the efficacy and safety of drugs. Gender-related differences have been shown for the metabolism and the effects of drugs. Gender differences have been described especially for the enzymes of the cytochrome P 450 family, but also for phase II reactions and most recently for P-glycoprotein. Most of these differences observed are only of minor clinical relevance, however may result in an increased rate of adverse drug reactions. Further differences may be based on different receptor/target sensitivities, e. g. the increased sensitivity for drug-induced torsade de pointes arrhythmia in women. In addition, in complex diseases such as heart failure, men and women may develop different mechanisms of counter- regulation requiring different therapeutic approaches. Population-based approaches demonstrate gender differences in the incidence of adverse drug reactions. PMID- 15887064 TI - [Clinical trials using antidepressants and antipsychotics. The pros and cons of placebo control]. AB - Controlled, randomized, double-blind, parallel group clinical trials are needed to establish efficacy and safety of new psychopharmacological drugs. Usually this is done by clinical trials including placebo control. These placebo-controlled trials have been criticized as being unethical in clinical situations where effective and acceptable treatment options are available. It has been argued that studies with potential new antidepressants or antipsychotics should employ only a comparator-controlled design, whereby new drugs have to be non-inferior or superior to existing treatment. In general actively controlled designs require the inclusion of larger patient populations. However, sole acceptance of "superior" compounds will hinder the development of more efficacious and better tolerated drugs. Moreover, acceptance of non-inferiority designs in psychopharmacology will be associated with the risk of ineffective compounds being approved. Due to these methodological reasons placebo-controlled studies of new antidepressants and antipsychotics are still justified and necessary, both ethically and scientifically. Therefore, from a regulatory point of view three arm studies including standard, new drug and placebo represent the preferred study design, which allows the most comprehensive interpretation of study results. However, measures should ensure that patients are not harmed by the act of forgoing established treatment options and that they provide fully informed consent. The article reviews the methodological and ethical rationale for conducting placebo-controlled trials with antidepressants and antipsychotics. PMID- 15887065 TI - [New guidelines for the assessment of bioavailability and bioequivalence]. AB - Bioavailability and bioequivalence studies are essential in the clinical development of medicinal products and the optimization of pharmaceutical forms. Bioavailability means the rate and extent to which the active substance or active moiety is absorbed from a pharmaceutical form and becomes available at the site of action. In practice, drug concentration-time courses are measured in the systemic circulation, and the area under the curve (AUC) as well as the observed maximum concentration (C(max)) are determined. Products are considered bioequivalent if their bioavailabilities after administration of the same molar doses are similar to such a degree that their effects, with respect to both efficacy and safety, will be essentially the same and thus, there are no relevant differences in terms of AUC and C(max). In 2002 a revised version of the 'Note for Guidance on the Investigation of Bioavailability and Bioequivalence' came into effect (CPMP/EWP/QWP/1401/98). Relevant changes in comparison to the previous version are: request for GLP-compliant bioanalytical measurements; for long half-life drugs a truncated AUC is acceptable; acceptance criteria for bioequivalence assessment and requirements for a waiver of bioequivalence studies were further specified. In this context the Biopharmaceutics Classification System (BCS) seems appropriate to decide whether in special cases of rapidly dissolving solid oral dosage forms a biowaiver may be granted or not. Products not considered critical in this matter are medicinal products for which the formulation does not affect the rate and extent of absorption, i. e. bioavailability, of the active moiety. Highly soluble (and highly permeable) drugs (BCS class I) are such candidates. Comprehensive state-of-the-art guidance on the design, conduct and analysis of bioavailability and bioequivalence studies is given in the current European guideline. PMID- 15887066 TI - [Regulatory aspects of clinical trials for medical devices and in vitro diagnostics]. AB - The European Medical Device Directives and In Vitro Diagnostic Directive were transposed into the German Medical Device Law (MPG) in 2002. Clinical trials in Germany are to be conducted in accordance with the MPG, Harmonized European Standards, professional codes and laws and for medical and scientific reasons according to ICH GCP. PMID- 15887067 TI - [Biometrical methods for the proof of efficacy in regulatory submissions. Non inferiority in clinical studies]. AB - The increasing number of approved efficacious therapies for various indications raises the question of whether the inclusion of a placebo group is still justified. In addition, pharmaceutical companies and regulatory agencies acknowledge that it may be sufficient to prove that a new therapy is comparable to an approved reference therapy regarding efficacy and safety in some situations. This becomes especially striking for the approval to market a generic drug. Another, perhaps even more important example is the increasing resistance of bacteria which calls for new antibiotics based on new therapeutic principles without having the claim for better efficacy. In these situations, a comparable efficacy would constitute progress. In the present paper, we discuss the numerous methodological challenges and approaches to overcome these problems that occur when it is not possible or even not wanted to use the classic approach of a randomized placebo-controlled superiority trial. Here, the field of medical biometry, which has proved in the last 25 years to be an integral part of the development of new drugs, demonstrates its suitability as a flexible and scientifically based means to fulfil the requirements resulting from clinical practice. Starting from the fact that statistical methods are not able to prove "equality" of two treatments, "shifted" hypotheses are considered and their importance for the different study designs is discussed. We show how the classic hypotheses known from placebo-controlled clinical trials can be embedded in this concept. The implications of this approach for the analysis and interpretation of study results is further discussed. The relevant guidelines of the European and US regulatory agencies are taken into account. PMID- 15887068 TI - Biostatistical methods for demonstrating efficacy in the regulatory setting. An epistemological approach to adaptive designs. AB - Adaptive designs are one of the most promising developments in statistics with applications to clinical trials. Obviously the knowledge at the beginning of a clinical trial will always be limited. In consequence it may be considered necessary that the knowledge from accumulating information should be used to optimize the design of the trial. This paper discusses conditions under which this may be possible in phase III clinical trials where the principle aim is confirmation of hypotheses that have been developed in earlier stages of drug development. PMID- 15887069 TI - [Post-marketing surveillance studies]. AB - Approval of a drug on the market is typically considered as the successful endpoint of a long development phase. This often leads to reduced interest in further research for the indication approved, while in contrast the patient's as well as physician's interest in further research increases. This paper discusses the main areas of effectiveness, safety and post-marketing practice regarding sensible study types for the specific area and the potential gain of knowledge. PMID- 15887070 TI - [The remarkable rise in life expectancy and how it will affect medicine]. AB - Life expectancy has increased at a steady pace in industrialized countries over the last 160 years. A slowdown is not evident: Since 1950 the number of people celebrating their 100th birthdays has at least doubled each decade. This increase in survival is the result of economic developments, social improvements and advances in medicine. Although the belief that old-age mortality is intractable remains widespread, life expectancy is not approaching a limit. Rather, the evidence suggests that ageing is plastic and that survival can be extended by various genetic changes and non-genetic interactions. Increases in life expectancy are largely attributed to improvements in old-age survival. It is a reasonable scenario that life expectancy will rise further in coming decades, supported by advances in the prevention, diagnosis and treatment of age-related diseases. If the trend continues, life expectancy in Germany will rise to over 90 years in the first half of this century. Many official forecasts, however, have assumed lower figures which can have severe consequences both for public and private decision making. PMID- 15887071 TI - [Important aspects of geriatric pharmacotherapy]. AB - Drug prescription for geriatric patients is one of the most prevalent yet also one of the most difficult interventions in geriatrics. Although this article focuses solely on specific important aspects of geriatric pharmacotherapy, medication in clinical practice is only one type of geriatric intervention that needs to be embedded in a well-coordinated bundle of multidimensional interventions addressing functional and psychosocial aspects alike. Not every older patient is per se a geriatric patient, nor is each geriatric patient per se of very old age. Therefore, this article initially proposes a clinical definition of the geriatric patient before summarizing the most significant physiological age changes affecting pharmacokinetics and pharmacodynamics as well as summarizing the most prevalent risks associated with the age-correlated increase in multiple drug use in order to describe the background from which general principles of geriatric pharmacotherapy have evolved. The quality of geriatric pharmacotherapy must not only be determined by avoidance of any kind of over medication, but also by the avoidance of under medication as well as the avoidance of inappropriate drugs for elderly patients. Treatment failures do not only result from mere polypharmacy, but can also be due to under utilization of drugs as well as due to the prescription of inappropriate medications. In fact, the problem of under medication in certain age-correlated diseases (e. g. depression, dementia, pain) has rarely been addressed systematically in clinical or epidemiological studies until now. In order to improve quality and safety of geriatric pharmacotherapy, a group of experts in the field developed a set of explicit age-adjusted criteria for potentially inappropriate drugs in 1991. These criteria have been regularly updated thereafter until 2003 and focus on age related aspects of specific drugs and dosages as well as on co-morbid states and co-medications. As long as geriatric patients are rarely included in clinical trials on drug effectiveness and drug safety, these criteria should be applied and used in clinical practice at least to identify any demand for a detailed explicit justification of a therapeutic decision not in line with the explicit criteria in order to enhance safety of drug treatment in geriatric patients. PMID- 15887072 TI - [Essentials of prevention policy in Germany]. AB - Prevention has become a major task of German health policy. In light of decreasing birth rates and low ages for starting retirement, prevention can contribute to stabilizing the social system by helping to prevent the outburst of diseases, ensure early diagnosis, and facilitate better coping strategies. It can also help to postpone or prevent early retirement and nursing care. To utilize preventive potentials German health policy has initiated the German Forum on Prevention and Health Promotion and a Law on Prevention, which is being negotiated in the German Parliament at present. In order to strengthen prevention, a system will be created that is financed by health, pension, accident and nursing insurance providers together. There will be campaigns, initiation of behavioural changes and setting activities (as required and described by WHO). The system will be controlled by common goals and targets, quality standards and a reporting duty to monitor the efficiency. On the federal level there will be a foundation on prevention and health promotion by the social insurance for the purpose of coordination. The German Forum has existed for 3 years now being a voluntary joint venture of relevant actors in prevention. The aim is to work together and create synergies. At present it is working on four topics: healthy kindergarden and schools, health promotion in firms, healthy aging and organisation and law. PMID- 15887073 TI - [The new human mode of being demented]. AB - Today, for the first time in the history of mankind, the number of demented people is so large that we are compelled to speak of a particular population group and a characteristic human mode of being. Ten perspectives describe which personal and societal changes will enable us to adjust to this new situation. Particular consideration is given to a new, more complete image of man. PMID- 15887074 TI - [The EU Bathing Water Directive. Risk assessment and standards]. AB - In an attempt to reduce the risk of infection in natural bathing waters the European Union is in the process of improving the Bathing Water Directive 76/160/EWG, which regulates the safety of such waters. The proposal contains several positive innovations which will improve the protection of the bathers: (1) health-related indicators, (2) harmonized detection methods, (3) requirements for active bathing water management, and (4) stricter standards for coastal waters. One of the most salient features of the current draft is the introduction of bacterial standards that are more stringent for coastal than for fresh waters. This decision on different standards seems unjustified: it was taken solely on the grounds that in two epidemiological studies-one carried out in coastal, the other in fresh waters-the maximum excess rate of gastroenteritis among bathers in coastal waters was higher than among bathers in fresh waters. However, it was not taken into account that the concentrations of bacterial indicators at which the gastroenteritis rate began to increase was nearly identical in both studies. The ratio between the standard concentrations of E. coli and intestinal enterococci in the draft was set at 2.5. This value does not correspond to the ratio found in German surface waters with low pollution levels, with ratios ranging from 2.7 to 4.0, and to the even higher ratios found in raw and treated sewage effluents. As a consequence in a majority of cases the non-compliance of bathing waters in Germany would be caused exclusively by a violation of E. coli standards. In assessing risks of infection it must also be taken into account that the adequacy of E. coli and intestinal enterococci for signaling the presence of viruses in water is far from optimal. The decay of viruses in water-estimated by the decay of bacteriophages-was found to be substantially slower than the die-off of indicator bacteria. PMID- 15887075 TI - [Standardization of substance contents in urine--creatinine. Statement of the Commission "Human Biomonitoring" of the Environmental Agency]. PMID- 15887077 TI - Changes in gene expression following high-temperature adaptation in experimentally evolved populations of E. coli. AB - Transcription profiling (quantitative analysis of RNA abundance) can provide a genome-wide picture of gene expression changes that accompany organismal adaptation to a new environment. Here, we used DNA microarrays to characterize genome-wide changes in transcript abundance in three replicate lines of the bacterium E. coli grown for 2,000 generations at a stressful high temperature (41.5 degrees C). Across these lines, 12% of genes significantly changed expression during high-temperature adaptation; the majority of these changes (55%) were less than twofold increments or decrements. Thirty-nine genes, four times the number expected by chance alone, exhibited moderately or highly replicated expression changes across lines. Expression changes within a priori defined functional categories showed an even greater level of replication than did individual genes. Expression changes in the phenotypically defined stress genes and adaptation functional categories were important in evolutionary adaptation to high temperature. PMID- 15887078 TI - Increased capacity for sustained locomotion at low temperature in parthenogenetic geckos of hybrid origin. AB - The evolution of parthenogenesis is typically associated with hybridization and polyploidy. These correlates of parthenogenesis may have important physiological consequences that need be taken into account in understanding the relative merits of sexual and parthenogenetic reproduction. We compared the thermal sensitivity of aerobically sustained locomotion in hybrid/triploid parthenogenetic races of the gecko Heteronotia binoei and their diploid sexual progenitors. Endurance times at low temperature (10 degrees , 12.5 degrees , and 15 degrees C, 0.05 km h(-1)) were significantly greater in parthenogenetic females than in sexual females. Comparison of oxygen consumption rates during sustained locomotion at increasing speeds (0.05, 0.10, 0.15, 0.20, 0.25, and 0.30 km h(-1), 25 degrees C) indicated that parthenogenetic lizards have higher maximum oxygen consumption rates and maximum aerobic speeds than do female sexual geckos. In addition, parthenogenetic geckos showed greater levels of voluntary activity at 15 degrees C than did sexual geckos, although this pattern appears strongest in comparison to male sexual forms. Parthenogenetic lineages of Heteronotia thus have an advantage over sexual lineages in being capable of greater aerobic activity. This result is opposite of that found in prior studies of parthenogenetic teiid lizards (genus Cnemidophorus) and highlights the idiosyncratic nature of phenotypic evolution in parthenogens of hybrid origin. PMID- 15887079 TI - Circulating catecholamines and cardiorespiratory responses in hypoxic lungfish (Protopterus dolloi): a comparison of aquatic and aerial hypoxia. AB - Circulating catecholamine levels and a variety of cardiorespiratory variables were monitored in cannulated bimodally breathing African lungfish (Protopterus dolloi) exposed to aquatic or aerial hypoxia. Owing to the purported absence of external branchial chemoreceptors in lungfish and the minor role played by the gill in O2 uptake, it was hypothesized that plasma catecholamine levels would increase only during exposure of fish to aerial hypoxia. The rapid induction of aquatic hypoxia (final PWo2 = 25.9+/-1.6 mmHg) did not affect the levels of adrenaline (A) or noradrenaline (NA) within the plasma. Similarly, none of the measured cardiorespiratory variables--including heart rate (fH), blood pressure, air-breathing frequency (fV), O2 consumption (Mo2), CO2 excretion (Mco2), or blood gases--were influenced by acute aquatic hypoxia. In contrast, however, the rapid induction of aerial hypoxia (inspired Po2=46.6+/-3.3 mmHg) caused a marked increase in the circulating levels of A (from 7.9+/-2.0 to 18.8+/-6.1 nmol L(-1)) and NA (from 7.7+/-2.2 to 19.7+/-6.3 nmol L(-1)) that was accompanied by significant decreases in Mo2, arterial Po2 (Pao2), and arterial O2 concentration (Cao2). Air-breathing frequency was increased (by approximately five breaths per hour) during aerial hypoxia and presumably contributed to the observed doubling of pulmonary Mco2 (from 0.25+/-0.04 to 0.49+/-0.07 mmol kg(-1) h(-1)); fH and blood pressure were unaffected by aerial hypoxia. An in situ perfused heart preparation was used to test the possibility that catecholamine secretion from cardiac chromaffin cells was being activated by a direct localized effect of hypoxia. Catecholamine secretion from the chromaffin cells of the heart, while clearly responsive to a depolarizing concentration of KCl (60 mmol L(-1)), was unaffected by the O2 status of the perfusion fluid. The results of this study demonstrate that P. dolloi is able to mobilize stored catecholamines and increase f(V) during exposure to aerial hypoxia while remaining unresponsive to aquatic hypoxia. Thus, unlike in exclusively water-breathing teleosts, P. dolloi would appear to rely solely on internal/airway O2 chemoreceptors for initiating catecholamine secretion and cardiorespiratory responses. PMID- 15887080 TI - High swimming and metabolic activity in the deep-sea eel Synaphobranchus kaupii revealed by integrated in situ and in vitro measurements. AB - Several complementary studies were undertaken on a single species of deep-sea fish (the eel Synaphobranchus kaupii) within a small temporal and spatial range. In situ experiments on swimming and foraging behaviour, muscle performance, and metabolic rate were performed in the Porcupine Seabight, northeast Atlantic, alongside measurements of temperature and current regime. Deep-water trawling was used to collect eels for studies of animal distribution and for anatomical and biochemical analyses, including white muscle citrate synthase (CS), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), and pyruvate kinase (PK) activities. Synaphobranchus kaupii demonstrated whole-animal swimming speeds similar to those of other active deep-sea fish such as Antimora rostrata. Metabolic rates were an order of magnitude higher (31.6 mL kg(-1) h(-1)) than those recorded in other deep-sea scavenging fish. Activities of CS, LDH, MDH, and PK were higher than expected, and all scaled negatively with body mass, indicating a general decrease in muscle energy supply with fish growth. Despite this apparent constraint, observed in situ burst or routine swimming performances scaled in a similar fashion to other studied species. The higher-than-expected metabolic rates and activity levels, and the unusual scaling relationships of both aerobic and anaerobic metabolism enzymes in white muscle, probably reflect the changes in habitat and feeding ecology experienced during ontogeny in this bathyal species. PMID- 15887081 TI - Factorial aerobic scope is independent of temperature and primarily modulated by heart rate in exercising Murray cod (Maccullochella peelii peelii). AB - Several previous reports, often from studies utilising heavily instrumented animals, have indicated that for teleosts, the increase in cardiac output (Vb) during exercise is mainly the result of an increase in cardiac stroke volume (V(S)) rather than in heart rate (fH). More recently, this contention has been questioned following studies on animals carrying less instrumentation, though the debate continues. In an attempt to shed more light on the situation, we examined the heart rates and oxygen consumption rates (Mo2; normalised to a mass of 1 kg, given as Mo2kg) of six Murray cod (Maccullochella peelii peelii; mean mass+/-SE = 1.81+/-0.14 kg) equipped with implanted fH and body temperature data loggers. Data were determined during exposure to varying temperatures and swimming speeds to encompass the majority of the biological scope of this species. An increase in body temperature (Tb) from 14 degrees C to 29 degrees C resulted in linear increases in Mo2kg (26.67-41.78 micromol min(-1) kg(-1)) and fH (22.3-60.8 beats min(-1)) during routine exercise but a decrease in the oxygen pulse (the amount of oxygen extracted per heartbeat; 1.28-0.74 micromol beat(-1) kg(-1)). During maximum exercise, the factorial increase in Mo2kg was calculated to be 3.7 at all temperatures and was the result of temperature-independent 2.2- and 1.7-fold increases in fH and oxygen pulse, respectively. The constant factorial increases in fH and oxygen pulse suggest that the cardiovascular variables of the Murray cod have temperature-independent maximum gains that contribute to maximal oxygen transport during exercise. At the expense of a larger factorial aerobic scope at an optimal temperature, as has been reported for species of salmon and trout, it is possible that the Murray cod has evolved a lower, but temperature-independent, factorial aerobic scope as an adaptation to the largely fluctuating and unpredictable thermal climate of southeastern Australia. PMID- 15887082 TI - Survival and physiological responses of hatchling blanding's turtles (Emydoidea blandingii) to submergence in normoxic and hypoxic water under simulated winter conditions. AB - Overwintering habits of hatchling Blanding's turtles (Emydoidea blandingii) are unknown. To determine whether these turtles are able to survive winter in aquatic habitats, we submerged hatchlings in normoxic (155 mmHg Po2) and hypoxic (6 mmHg Po2) water at 4 degrees C, recording survival times and measuring changes in key physiological variables. For comparison, we simultaneously studied hatchling softshell (Apalone spinifera) and snapping (Chelydra serpentina) turtles, which are known to overwinter in aquatic habitats. In normoxic water, C. serpentina and A. spinifera survived to the termination of the experiment (76 and 77 d, respectively). Approximately one-third of the E. blandingii died during 75 d of normoxic submergence, but the cause of mortality was unclear. In hypoxic water, average survival times were 6 d for A. spinifera, 13 d for E. blandingii, and 19 d for C. serpentina. Mortality during hypoxic submergence was probably caused by metabolic acidosis, which resulted from accumulated lactate. Unlike the case with adult turtles, our hatchlings did not increase plasma calcium and magnesium, nor did they sequester lactate within the shell. Our results suggest that hatchling E. blandingii are not particularly well suited to hibernation in hypoxic aquatic habitats. PMID- 15887083 TI - Steroid hormone metabolism by the chorioallantoic placenta of the mountain spiny lizard Sceloporus jarrovi as a possible mechanism for buffering maternal-fetal hormone exchange. AB - The placenta provides a maternal-fetal exchange interface that maximizes the diffusion of gases, nutrients, and wastes. However, the placenta also may permit diffusion of lipid-soluble steroid hormones that influence processes such as sex specific fetal development and maternal pregnancy maintenance. In mammals, placental steroid metabolism contributes to regulation of maternal and fetal hormone levels. Such mechanisms may be less highly developed in species that have recently evolved placentation, such as many reptiles. We therefore chose to investigate placental metabolism of steroids in the viviparous lizard Sceloporus jarrovi. In vitro tissue incubations tested the abilities of the chorioallantoic placenta to clear progesterone and corticosterone by converting them to other metabolites and to synthesize progesterone. Placental tissue rapidly cleared progesterone and corticosterone added to the incubation media, indicating that the tissue had converted the steroids to other products. Placental tissue also synthesized substantial concentrations of progesterone from the prohormone pregnenolone. Thus, even in a species with a simple, recently evolved placenta, steroid metabolism appears to be highly developed and could be critical for regulation of maternal and fetal hormone levels. This finding suggests that placental hormone metabolism may be critical to the successful evolution of placentation. PMID- 15887084 TI - Extracellular and intracellular acid-base status with regard to the energy metabolism in the oyster Crassostrea gigas during exposure to air. AB - The acid-base status of extra- and intracellular fluids was studied in relation to the anaerobic energy metabolism in the adductor muscle, mantle, gills, and heart of the marine bivalve Crassostrea gigas after exposure to air for periods of 2, 4, 8, 12, 24, and 48 h. Such exposure was found to cause a significant reduction in the pH in the hemolymph (pH(e)) within the first 4 h. The decrease in the pHe was accompanied by elevated Pco2 values, causing [HCO3-] to rise (respiratory acidosis). Thereafter, the pHe fell at a lower rate, and this fall was partially compensated for by a further increase in [HCO3-] in the hemolymph. The increase in the [Ca] levels in the hemolymph indicates a mobilization of Ca2+ from CaCO3 and the involvement of bicarbonates in the buffering of pHe. The main anaerobic end-products that accumulated in the tissues during the first stages of anaerobiosis were alanine and succinate, at a ratio of about 2 : 1. Later on, propionate and acetate were also accumulated at significant rates. In contrast to the adductor muscle, gills, and mantle, opine production in the heart was significant after 12-24 h of exposure to air. Determination of intracellular pH (pHi) revealed that there is a close relationship between the rate of anaerobic end-product accumulation and the extent of intracellular acidosis in the adductor muscle, mantle, and gills. On the contrary, accumulation of anaerobic end products in the heart did not cause any significant change in its pHi. The intracellular nonbicarbonate, nonphosphate buffering value (beta (NB,NPi)) was determined to be higher in the heart than in the other three tissues and thus probably plays a crucial role in stabilizing heart pHi during exposure to air. PMID- 15887085 TI - Oxygen preference of Daphnia magna is influenced by Po2 acclimation and biotic interactions. AB - Oxygen preference of the planktonic crustacean Daphnia magna was observed within an oxygen gradient. Long-term acclimation to specific oxygen conditions determined the preferred oxygen tension. Animals from hypoxic acclimation stayed in oxygen-poor water at first. They chose oxygen tensions above the critical partial pressure, thus maintaining aerobic energy supply, but they avoided higher oxygen concentrations. However, they gradually took more oxygen-rich positions within 2 d. Animals from severe hypoxic and hyperoxic acclimation showed stenoxic behavior. Specimens raised in moderately hypoxic or normoxic medium spread out more widely in the oxygen gradient (euryoxic behavior). The possible role of hemoglobin and antioxidant defense capacity limiting the animals' distribution at low or high oxygen concentrations, respectively, is discussed. In tests with mixed groups of D. magna from opposing oxygen acclimation conditions, the animals clustered at intermediate oxygen conditions. However, the acclimation condition of the major group was found to determine the position taken within the gradient to a greater extent. Selected oxygen tensions were closer to the preference values of the larger group, yet the influence of the minority prevented the majority in heterogeneous groups from taking their originally determined preference position. Thus, aggregation behavior interfered with the acclimation dependent oxygen preference behavior. PMID- 15887086 TI - Cutaneous blood flow and water absorption by dehydrated toads. AB - Blood cell flux (BCF) in ventral pelvic skin capillaries was measured in toads, Bufo woodhouseii and Bufo punctatus, using a chamber that allowed hydration behavior and water absorption to be observed concurrently in unrestrained animals. Dehydrated B. woodhouseii and B. punctatus placed on a rehydration solution significantly increased BCF relative to that on a dry surface in less than 2 min. Skin contact with a rehydration solution rather than dehydration alone is the primary stimulus for increased seat patch blood flow. In B. woodhouseii, the water absorption response was initiated after the increase in BCF had started but before maximum BCF was reached. BCF and water uptake across the ventral skin of both species placed on deionized water were not different from those of toads placed on 50 mM NaCl. Similarly, no significant correlation between BCF and rate of water uptake could be observed in dehydrated toads of either species. Angiotensin II (AII) injection in hydrated B. punctatus had no effect on BCF, suggesting that factors other than AII are responsible for the increase in blood flow upon water contact in dehydrated toads. PMID- 15887087 TI - Humic substances influence sodium metabolism in the freshwater crustacean Daphnia magna. AB - Humic substances are ubiquitous components of natural waters with important roles in alleviating metal toxicity to aquatic organisms. Recent literature reports suggest that humic substances may also exert direct influences on biota. This study investigated the influence of two commercially available humic substances on sodium metabolism in Daphnia magna, a hyperregulating freshwater crustacean. Environmentally realistic levels of Suwannee River natural organic matter (SRN) and Aldrich humic acid (AHA) significantly enhanced sodium transport. This effect was described as an uncompetitive stimulation of sodium influx, as characterised by an increased maximal sodium transport rate (Jmax), accompanied by a decreased uptake affinity (increased Km). SRN exposure also significantly promoted the unidirectional loss of sodium from the daphnids to the water, an effect not observed in the presence of AHA. A 24-h preexposure to AHA before influx measurement had no effect on AHA-induced stimulation of sodium influx. Conversely, 24-h preexposure to SRN resulted in influx values that returned to control (humic-free) levels. Whole-body sodium levels reduced by SRN exposure were also restored to control levels following 24-h SRN preexposure. The significance and potential mechanisms of these actions are discussed, and the toxicological implications of these findings are assessed. PMID- 15887088 TI - The energetics of trading nuptial gifts for copulations in katydids. AB - During copulation, male Isophya kraussi transfer a large nuptial gift to females. In this study, we hypothesized that the energy content of spermatophores should meet the energy requirements of both body maintenance and egg production of females. We measured the field metabolic rate of male and female I. kraussi using the doubly labeled water method and the energy content of spermatophores and male bodies with microbomb calorimetry. The energy content of male nuptial gifts averaged 0.66+/-0.09 kJ, approximately 20% of the total body energy content of male I. kraussi (3.24+/-0.26 kJ). Field metabolic rates averaged 0.41+/-0.17 kJ d(-1) (n = 8) for males and 0.30+/-0.15 kJ d(-1) (n = 5) for female I. kraussi. Thus, the energy content of spermatophores exceeded the daily energy requirements of existence in male I. kraussi. A single nuptial gift provides for all energy requirements of females for 1 or 2 d, depending on their activity, egg production, and ambient temperature. Because the shortest known remating interval of female katydids varies between 1 and 3 d, female I. kraussi could theoretically exist exclusively on spermatophores to meet their nutritional requirements. PMID- 15887089 TI - A cholesterol-enriched diet enhances egg production and egg viability without altering cholesterol Content of biological membranes in the copepod Acartia hudsonica. AB - Copepods may lack the capacity for de novo synthesis of cholesterol, while at the same time their dietary levels of sterol vary. We tested the hypothesis that copepods maintain the cholesterol contents of their biological membranes despite varying dietary levels of cholesterol. Acartia hudsonica were acclimated for 5 d to phytoplankton alone or phytoplankton supplemented with cholesterol, at a level sufficient to induce a maximal response on egg production rates. Biological membranes were prepared from the copepods and cholesterol contents assayed. Egg production and hatch rates were measured (the former to confirm that supplemented cholesterol was being assimilated). Analyses of marker enzymes indicate that the majority of membrane-associated cholesterol in the copepod resides in the plasma membrane. In membranes fractions, cholesterol normalized to protein or activity of Na+/K+-ATPase is not significantly different for supplemented and unsupplemented groups (29 and 33 mu g cholesterol mg(-1) protein, respectively; 0.24 and 0.25 mg cholesterol U(-1) Na+/K+-ATPase, respectively). At the same time, acclimating animals to a diet enriched with cholesterol enhances egg production by up to 1.8-fold and egg viability by 1.5-fold. We conclude that a cholesterol-enriched diet stimulates both egg production and hatching rates without altering cholesterol contents of plasma membranes in the copepod A. hudsonica. PMID- 15887090 TI - Digestive organ sizes and enzyme activities of refueling western sandpipers (Calidris mauri): contrasting effects of season and age. AB - We examined seasonal and age-related variation in digestive organ sizes and enzyme activities in female western sandpipers (Calidris mauri) refueling at a coastal stopover site in southern British Columbia. Adult sandpipers exhibited seasonal variation in pancreatic and intestinal enzyme activities but not in digestive system or organ sizes. Spring migrants had 22% higher total and 67% higher standardized pancreatic lipase activities but 37% lower total pancreatic amylase activity than fall migrants, which suggests that the spring diet was enriched with lipids but low in glycogen. Spring migrants also had 47% higher total intestinal maltase activity as well as 56% higher standardized maltase and 13% higher standardized aminopeptidase-N activities. Spring migrants had higher total enzymic capacity than fall migrants, due primarily to higher total lipase and maltase activities. During fall migration, the juvenile's digestive system was 10% larger than the adult's, and it was composed differently: juveniles had a 16% larger small intestine but a 27% smaller proventriculus. The juvenile's larger digestive system was associated with lower total enzymic capacity than the adult's due to 20% lower total chitinase and 23% lower total lipase activities. These results suggest that juvenile western sandpipers may process food differently from adults and/or have a lower-quality diet. PMID- 15887091 TI - Redistribution of extracellular water and sodium may contribute to saline tolerance in wild ducks. AB - The compartmentalization of body fluids was measured in three species of ducks that differ in saline tolerance. Half of the birds of each species drank freshwater, while the other half drank saline (300 mM NaCl). Among ducks that drank freshwater, total body water (TBW) was similar among all species, but Barrow's goldeneyes (Bucephala islandica), the most marine species, had larger extracellular fluid volume (ECFV) than freshwater mallards (Anas platyrhynchos) or estuarine canvasbacks (Aythya valisineria). When acclimated to saline, only goldeneyes shifted extracellular water and Na+ into the intracellular compartment. ECFV was correlated with plasma aldosterone concentration in goldeneyes, but not in canvasbacks (aldosterone was not measured in mallards). Data summarized from the literature showed that TBW does not differ among terrestrial, freshwater, or marine species, but marine species have a larger part of their TBW in the extracellular compartment. Saline induced movement of extracellular water and Na+ into the cells only in goldeneyes. ECFV and redistribution of extracellular water and Na+ into the cells may be important components in saline tolerance of marine birds. PMID- 15887092 TI - Nitrogen requirements and urea recycling in an omnivorous marsupial, the northern brown bandicoot Isoodon macrourus. AB - The maintenance nitrogen (N) requirement of the northern brown bandicoot, Isoodon macrourus, was determined by giving captive animals diets based on a commercial small carnivore mix and shredded sweet potato, with 20% sand included in the moist diet to simulate the soil ingested by bandicoots during natural foraging. Linear regression of N balance on N intake yielded maintenance N requirements of 438 mg kg(-0.75) d(-1) on a dietary basis and 413 mg kg(-0.75) d(-1) on a truly digestible basis. These values are high when compared with the range reported for other marsupial omnivores (46-146 mg kg(-0.75) d(-1)) and the omnivorous eutherian mouse (265 mg kg(-0.75) d(-1)). Possible reasons include an energy deficiency on the low and medium N diets that could have resulted in an overestimate of up to 45%, relative deficiencies of methionine and isoleucine in all diets (but the effect of these is difficult to quantify), high metabolic fecal nitrogen (MFN) due to the abrasive effect of sand on the digestive tract mucosa (but MFN was no lower without sand in the diets), or high rates of N loss in both feces and urine as seen in eutherian carnivores. The northern brown bandicoot is perhaps the most insectivorous of all peramelid marsupials, suggesting that the high maintenance requirements for N are an intrinsic characteristic of the species. Limited recycling of endogenously produced urea to the gut, only 28% on the low N diet, is consistent with this conclusion. PMID- 15887093 TI - Mapping of a single locus capable of complementing the defective heterochromatin phenotype of Roberts syndrome cells. AB - Roberts syndrome (RS) is a developmental disorder characterized by tetraphocomelia and a broad spectrum of additional clinical features. Most patients with RS exhibit characteristic cytogenetic phenotypes, which include an abnormal appearance of pericentromeric heterochromatin on metaphase chromosomes, referred to as "heterochromatic repulsion." In the present study, we use complementation of this abnormal cytogenetic phenotype as a means to identify a specific region of the normal human genome capable of rendering phenotypic correction. We screened the entire human genome, using a transient chromosome transfer assay, and demonstrated complementation exclusively after the transfer of proximal chromosome 8p, a result subsequently confirmed by stable microcell mediated chromosome transfer. Additionally, homozygosity mapping was used to refine the interval of this complementing locus to 8p21. The results are consistent with the notion that the single gene defect responsible for heterochromatic splaying and developmental abnormalities maps to chromosome 8p21. PMID- 15887094 TI - An in vivo microfabricated scaffold for tendon repair. AB - A new type of in vivo tissue engineering system for tendon repair in situ after cut or crush of a flexor tendon is described. The system is based on the topographical reaction, alignment, migration and perhaps proliferation of tendon cells on micrometrically grooved substrates made in a biodegradable polymer. Macrophage trapping in the structure may also help to prevent inflammation. Tendon damage including crush and section injury is a fairly frequent occurrence. The conventional treatment is surgical repair, however frequently this leads, especially in hand wounds, to attachment of the tendon surface to the surrounding synovium, which is very undesirable. We present an approach based on using a biodegradable device to ensure that the healing of severed or crushed flexor tendons is aided, synovial adhesion prevented and the final result anatomically correct. The biodegradable sheath carries microgrooves fabricated into the polymer by embossing that orient and guide the cells towards each other from either side of the region of damage. After six weeks an apparently normal functional tendon is reformed. PMID- 15887097 TI - Quality of care in gastroenterology: beyond the bell curve. PMID- 15887098 TI - Image of the month. Choledochocele type 3 choledochal cyst. PMID- 15887099 TI - Immunohistochemical analysis reveals high frequency of PMS2 defects in colorectal cancer. AB - BACKGROUND & AIMS: Germline mutations in the DNA mismatch repair (MMR) genes MSH2, MSH6, or MLH1 predispose to colorectal cancer (CRC) with an autosomal dominant inheritance pattern. The protein encoded by PMS2 is also essential for MMR; however, alterations in this gene have been documented only in extremely rare cases. We addressed this unexpected finding by analyzing a large series of CRCs. METHODS: Expression of MSH2, MSH6, MLH1, and PMS2 was studied by immunohistochemistry in 1048 unselected, consecutive CRCs. Where absence of MMR proteins was detected, microsatellite instability and cytosine methylation of the respective gene promoter were analyzed. The DNA of patients presenting with PMS2 deficient cancers was examined for germline and somatic alterations in the PMS2 gene. RESULTS: An aberrant pattern of MMR protein expression was detected in 13.2% of CRCs. Loss of expression of MSH2, MSH6, or MLH1 was found in 1.4%, 0.5%, and 9.8%, respectively. PMS2 deficiency accompanied by microsatellite instability was found in 16 cases (1.5%) with a weak family history of cancer. The PMS2 promoter was not hypermethylated in these cases. Despite interference of the PMS2 pseudogenes, we identified several heterozygous germline mutations in the PMS2 gene. CONCLUSIONS: PMS2 defects account for a small but significant proportion of CRCs and for a substantial fraction of tumors with microsatellite instability. However, the penetrance of heterozygous germline mutations in PMS2 is considerably lower than that of mutations in other MMR genes. The possible underlying causes of this unorthodox inheritance pattern are discussed. PMID- 15887101 TI - A quantitative analysis of NSAID-induced small bowel pathology by capsule enteroscopy. AB - BACKGROUND & AIMS: Conventional acidic nonsteroidal anti-inflammatory drugs frequently cause small bowel inflammation. Diagnosis is largely based on assay of surrogate markers of inflammation in stool, such as fecal calprotectin. However, stool markers are not widely available and the precise nature of this inflammation is uncertain. We used wireless capsule enteroscopy to quantitate and assess the nature of the small bowel damage caused by nonsteroidal anti inflammatory drugs when taken on a short-term basis. METHODS: Forty healthy volunteers underwent a baseline capsule enteroscopy and fecal calprotectin test. After taking diclofenac slow-release 75 mg twice a day (with omeprazole 20 mg twice a day for gastroprotection) for a total of 14 days, both investigations were repeated. RESULTS: After drug treatment, 30 subjects (75%) had increased repeat fecal calprotectin concentrations above the upper limit of normal. Capsule enteroscopy showed new pathology in 27 subjects (68%). The commonest lesions were mucosal breaks, seen in 16 (40%), which were seen to be bleeding in 2 (5%); reddened folds in 14 (35%); petechiae or red spots in 13 (33%); denuded mucosa in 8 (20%); and blood in the lumen without a visualized source in 3 (8%). Fifteen of the 27 subjects had more than one lesion concurrently. CONCLUSIONS: This study provides both biochemical and direct evidence of macroscopic injury to the small intestine in 68%-75% of volunteers resulting from 2 weeks' ingestion of slow release diclofenac. PMID- 15887102 TI - Asynchrony between the circular and the longitudinal muscle contraction in patients with nutcracker esophagus. AB - BACKGROUND & AIMS: The increases in intraluminal pressure and muscle cross sectional area (CSA) during esophageal contraction are markers of circular and longitudinal muscle contractions. The goal of our study was to determine temporal synchrony between circular and longitudinal muscle contraction in healthy subjects and patients with nutcracker esophagus. METHODS: Pressure and high frequency intraluminal ultrasound (HFIUS) images were recorded simultaneously in healthy subjects and patients with nutcracker esophagus at 2 and 10 cm above the lower esophageal sphincter during wet swallow. HFIUS images were digitized and analyzed for the muscle CSA. The time interval (delta-t) between the peak muscle CSA and the peak pressure was determined. RESULTS: In healthy subjects, a close temporal correlation existed between the peak contraction pressure and the peak muscle CSA with a maximum delta-t of 0.5 seconds at the 2- and 10-cm levels (0 0.5 seconds). On the other hand, the patient group had a median delta-t of 1.25 seconds (0.75-3.5 seconds) at the 2-cm level and 0.75 seconds (0-2.0 seconds) at the 10-cm level. Ninety-eight of 103 contractions in patients showed a delta-t >0.5 seconds. There was a significant correlation between delta-t and the amplitude of pressure wave, the duration of pressure wave, and the peak muscle CSA. The duration of pressure wave but not the duration of CSA wave was longer in patients with nutcracker esophagus as compared with healthy subjects. CONCLUSIONS: Patients with nutcracker esophagus show temporal asynchrony between the contractions of circular and longitudinal muscle layers. PMID- 15887103 TI - Precursor lesions in patients with multiple endocrine neoplasia type 1-associated duodenal gastrinomas. AB - BACKGROUND & AIMS: The identification of precursor lesions has a great impact on the understanding of tumorigenesis. Precursor lesions of endocrine tumors are known to occur in the setting of the MEN1 syndrome. The aim of this study was to test the hypothesis that MEN1-associated duodenal gastrinomas originate from diffuse preneoplastic gastrin cell changes. Precursor lesions may precede the development of duodenal gastrinomas because, in contrast to sporadic gastrinomas, these tumors are usually multiple. METHODS: The distribution of endocrine cells in the nontumorous duodenal tissue was analyzed qualitatively and quantitatively for 25 patients operated on for a duodenal gastrinoma. MEN1 status was assessed clinically and by polymerase chain reaction-based mutational analysis. RESULTS: Fourteen of 25 patients with gastrinoma had proliferative, hyperplastic lesions consisting of gastrin cells in the nontumorous duodenal mucosa, similar to the gastric enterochromaffin-like cell lesions observed in chronic atrophic gastritis. All patients with Zollinger-Ellison syndrome with proven MEN1 had such proliferative gastrin cell lesions, and all patients with Zollinger-Ellison syndrome without precursor lesions were MEN1 negative. CONCLUSIONS: Duodenal gastrinomas in MEN1, but not sporadic duodenal gastrinomas, are associated with proliferative gastrin cell changes within the nontumorous mucosa. It is likely that these lesions precede the development of MEN1-associated duodenal gastrinomas. PMID- 15887104 TI - Phenotypic variation in functional disorders of defecation. AB - BACKGROUND & AIMS: Although obstructed defecation is generally attributed to pelvic floor dyssynergia, clinical observations suggest a wider spectrum of anorectal disturbances. Our aim was to characterize phenotypic variability in constipated patients by anorectal assessments. METHODS: Anal pressures, rectal balloon expulsion, rectal sensation, and pelvic floor structure (by endoanal magnetic resonance imaging) and motion (by dynamic magnetic resonance imaging) were assessed in 52 constipated women and 41 age-matched asymptomatic women. Phenotypes were characterized in patients by principal components analysis of these measurements. RESULTS: Among patients, 16 had a hypertensive anal sphincter, 41 had an abnormal rectal balloon expulsion test, and 20 had abnormal rectal sensation. Forty-nine patients (94%) had abnormal pelvic floor motion during evacuation and/or squeeze. After correcting for age and body mass index, 3 principal components explained 71% of variance between patients. These factors were weighted most strongly by perineal descent during evacuation (factor 1), anorectal location at rest (factor 2), and anal resting pressure (factor 3). Factors 1 and 3 discriminated between controls and patients. Compared with patients with normal (n = 23) or reduced (n = 18) perineal descent, patients with increased (n = 11) descent were more likely (P < or = .01) to be obese, have an anal resting pressure >90 mm Hg, and have a normal rectal balloon expulsion test result. CONCLUSIONS: These observations demonstrate that functional defecation disorders comprise a heterogeneous entity that can be subcharacterized by perineal descent during defecation, perineal location at rest, and anal resting pressure. Further studies are needed to ascertain if the phenotypes reflect differences in the natural history of these disorders. PMID- 15887105 TI - Clinical, morphologic, and molecular features defining so-called telangiectatic focal nodular hyperplasias of the liver. AB - BACKGROUND & AIMS: Telangiectatic focal nodular hyperplasia (TFNH) of the liver is generally believed to belong to the focal nodular hyperplasia (FNH) family. The aim of this study was to use molecular markers, in addition to morphologic features, to better characterize TFNH. METHODS: Thirteen patients with TFNH were compared with 28 patients with FNH and 17 patients with hepatocellular adenoma. Full clinical and morphologic data were analyzed. Molecular markers included determination of clonality by examining the active X chromosome, genome-wide allelotyping, a search for hepatocyte nuclear factor 1alpha (HNF1alpha) mutations, and determination of ANGPT1/ANGPT2 transcript levels. RESULTS: No clinical differences were evident between patients with TFNH and adenoma; in particular, bleeding was observed in 77% and 53% of the cases, respectively. Patients with TFNH were more likely to experience nodule recurrence and the presence of multiple nodules than those with either FNH or adenoma. All TFNH and adenoma samples that were available for analysis were monoclonal, in contrast to 40% of the FNH samples. Chromosome losses confirmed monoclonality and were significantly less frequent in TFNH and FNH (22% and 26%) than in adenoma (53%). HNF1alpha mutations were found exclusively in half of the adenomas. ANGPT2 was overexpressed in TFNH and down-regulated in adenoma (P < .01) and FNH (P < .0005). CONCLUSIONS: TFNHs are monoclonal lesions frequently subject to bleeding that are similar to adenomas not carrying HNF1alpha mutations and require a similar type of treatment. However, morphologic and molecular data support the hypothesis that TFNH is a separate entity. PMID- 15887106 TI - Recombinant probiotics for treatment and prevention of enterotoxigenic Escherichia coli diarrhea. AB - BACKGROUND & AIMS: We have developed a therapeutic strategy for gastrointestinal infections that is based on molecular mimicry of host receptors for bacterial toxins on the surface of harmless gut bacteria. The aim of this study was to apply this to the development of a recombinant probiotic for treatment and prevention of diarrheal disease caused by enterotoxigenic Escherichia coli strains that produce heat-labile enterotoxin. METHODS: This was achieved by expressing glycosyltransferase genes from Neisseria meningitidis or Campylobacter jejuni in a harmless Escherichia coli strain (CWG308), resulting in the production of a chimeric lipopolysaccharide capable of binding heat-labile enterotoxin with high avidity. RESULTS: The strongest heat-labile enterotoxin binding was achieved with a construct (CWG308:pLNT) that expresses a mimic of lacto-N-neotetraose, which neutralized > or = 93.8% of the heat-labile enterotoxin activity in culture lysates of diverse enterotoxigenic Escherichia coli strains of both human and porcine origin. When tested with purified heat labile enterotoxin, it was capable of adsorbing approximately 5% of its own weight of toxin. Weaker toxin neutralization was achieved with a construct that mimicked the ganglioside GM2. Preabsorption with, or coadministration of, CWG308:pLNT also resulted in significant in vivo protection from heat-labile enterotoxin-induced fluid secretion in rabbit ligated ileal loops. CONCLUSIONS: Toxin-binding probiotics such as those described here have considerable potential for prophylaxis and treatment of enterotoxigenic Escherichia coli-induced travelers' diarrhea. PMID- 15887107 TI - Helicobacter pylori cag-type IV secretion system facilitates corpus colonization to induce precancerous conditions in Mongolian gerbils. AB - BACKGROUND & AIMS: Epidemiological studies suggest that atrophic corpus-dominant gastritis is an increased risk factor for gastric carcinogenesis. The role of the Helicobacter pylori type IV secretion system (T4SS) for pathogenesis in the Mongolian gerbil model was explored. METHODS: Mongolian gerbils were infected for 32 weeks either with H. pylori type I strain B128 or with isogenic mutant strain B128delta cytotoxin-associated gene (cagY) or B128delta cagA , defective in T4SS or in the production of its effector protein CagA, respectively. Quantitative H. pylori reisolation was performed from the gastric antrum and corpus separately, cytokines were measured by quantitative reverse-transcription polymerase chain reaction, and gastric pH and hormones were determined. RESULTS: B128-infected gerbils harbored high numbers of bacteria in the gastric antrum and corpus, whereas B128delta cagY and B128delta cagA colonized the antrum more densely than the corpus. All infected animals showed a strong antral inflammation and epithelial cell proliferation. B128-infected, rather than mutant-infected, gerbils presented a severe transmural inflammation with huge lymph aggregates, increased proliferation, significant atrophy, and mucous gland metaplasia in the corpus. Plasma gastrin levels and gastric pH values were significantly increased only in B128-infected gerbils. In all infected animals, the expression of the proinflammatory cytokines interleukin 1beta, interferon gamma, and growth regulated protein was considerably increased in the antrum, but only in wild type infected animals was an increase seen in the corpus mucosa. CONCLUSIONS: The presence of an intact T4SS allows H. pylori to colonize the gastric corpus. This results in atrophic corpus-dominant gastritis, a severe precancerous condition, thus highlighting T4SS and CagA as major risk factors for gastric cancer development. PMID- 15887108 TI - Different sensitivity of lamina propria T-cell subsets to nitric oxide-induced apoptosis explains immunomodulatory activity of a nitric oxide-releasing derivative of mesalamine in rodent colitis. AB - BACKGROUND & AIMS: Uncontrolled T-cell activation plays a critical role in the pathogenesis of inflammatory bowel diseases. Therefore, pharmacological strategies directed toward restoring the normal responsiveness of the immune system could be effective in the treatment of these pathologic conditions. The addition of a nitric oxide-releasing moiety to conventional drugs, such as aspirin and other anti-inflammatory analgesic drugs, results in new chemical entities with potent immunomodulatory activities. The aim of this study was to investigate the immunomodulatory activity of a nitric oxide-releasing derivative of mesalamine (NCX-456), as compared with standard mesalamine, in 2,4,6 trinitrobenzene sulfonic acid-induced colitis in mice. METHODS: Cells and tissues from mice with 2,4,6-trinitrobenzene sulfonic acid-induced colitis and from interleukin 10-deficient mice with spontaneous chronic colitis receiving treatment with several doses of NCX-456 or mesalamine were analyzed for morphology, cytokine production, and apoptosis. RESULTS: NCX-456, but not mesalamine, administration resulted in a marked reduction in clinical, histological, and immunologic signs of colitis in both models. NCX-456 inhibited the release of T-helper type 1-derived cytokines and increased the release of the regulatory T cell-derived cytokines interleukin 10 and transforming growth factor beta. In vitro analyses showed that NCX-456 inhibited proliferation and caused selective apoptosis of the subset of activated lamina propria T-helper type 1 cells, whereas it was ineffective for regulatory T-cell function and survival. CONCLUSIONS: Collectively, these data show that NCX-456 inhibits lamina propria T helper type 1 function and stimulates the activity of interleukin 10- and transforming growth factor beta-secreting cells, thus restoring mucosal immune homeostasis and suppressing intestinal inflammation. PMID- 15887109 TI - Interferon gamma induces translocation of commensal Escherichia coli across gut epithelial cells via a lipid raft-mediated process. AB - BACKGROUND & AIMS: The "leaky gut" hypothesis proposes that leakage of enteric bacteria into the body resulting from disruption of the epithelial barrier is a critical step in the pathophysiology of various disorders such as inflammatory bowel disease and sepsis. However, the pathways and underlying mechanisms by which commensal bacteria cross the epithelial barrier in inflammatory conditions remain unclear. This study investigated the mechanisms of interferon gamma mediated bacterial translocation across human colonic epithelial monolayers. METHODS: Caco-2 and T84 monolayers were exposed to interferon gamma. Barrier function was assessed by transepithelial electrical resistance and lucifer yellow permeability. Internalization and translocation of Escherichia coli strain C25 were measured by quantitative bacterial culture. Expression and distribution of junctional proteins were assessed by immunoblotting and confocal imaging. RESULTS: Minimal apical to basolateral translocation of C25 was observed in untreated T84 and Caco-2 monolayers. Interferon gamma caused a dramatic, dose dependent increase in C25 translocation, which was uncoupled from cytokine induced increases in paracellular permeability and disruption of tight junction proteins at low interferon gamma concentrations. These effects were associated with increased internalization of viable bacteria into, but not adherence to, Caco-2 cells. Interferon gamma-mediated bacterial translocation was abolished by pretreatment with the cholesterol-disrupting drugs filipin and methyl-beta cyclodextrin, whereas these agents had no effect on infection of Caco-2 by the enteric pathogen Shigella sonnei. CONCLUSIONS: Normally poorly invasive enteric bacteria may, in situations of inflammatory stress, exploit lipid raft-mediated transcytotic pathways to cross the intestinal epithelium, and these effects may precede cytokine-induced disruption of tight junctions. PMID- 15887110 TI - In vivo generation of oligoclonal colitic CD4+ T-cell lines expressing a distinct T-cell receptor Vbeta. AB - BACKGROUND & AIMS: Transplantation of wild-type (H-2k) bone marrow into tg epsilon26 mice (BM-->tg epsilon26) induces colitis, characterized by T-helper cell type 1 activation in the lamina propria. Here we determined whether pathogenic T-cell clones could be derived by serial adoptive transfers into healthy tg epsilon26 recipients, starting with the population of CD4+ cells in the mesenteric lymph nodes of BM-->tg epsilon26 mice. METHODS: CD4+ cells purified from the mesenteric lymph nodes of colitic BM-->tg epsilon26 mice were adoptively transferred into a second group of healthy tg epsilon26 recipients. Mesenteric lymph node CD4+ cells from the second group of mice were then used for consecutive transfers. Lamina propria CD4+ cells isolated from each mouse with colitis were analyzed for their cytokine profile and for their T-cell receptor Vbeta repertoire. RESULTS: CD4+ T cells maintained a dominant T-helper 1 phenotype after multiple transfers (< or = 8) into recipient tg epsilon26 mice. A single T-cell receptor Vbeta was enriched (as much as 90%) in 8 CD4+ T-cell lines: Vbeta8S3, Vbeta8S1/2, Vbeta10S1, or Vbeta14. Sequence analyses of the T cell receptor Vbetas showed clonality or the presence of a very restricted number of clones within each line. Adoptive transfers of the oligoclonal lines into either C3H x Rag-/- or severe combined immunodeficiency disease mice (H-2k) also induced colitis, whereas transfers into BALB/c x Rag-/- or severe combined immunodeficiency disease mice (H-2d) did not. CONCLUSIONS: Colitis-inducing CD4+ T-helper 1 cell clones can be obtained by enrichment through sequential adoptive transfers of CD4+ cells from mesenteric lymph nodes. Distinct dominant T-cell receptor Vbetas in each cell line responded to antigens presented by class II major histocompatibility complex. PMID- 15887111 TI - Tumor necrosis factor alpha blockade restores growth hormone signaling in murine colitis. AB - BACKGROUND & AIMS: Cytokines including tumor necrosis factor alpha (TNFalpha) may create a state of growth hormone (GH) resistance in Crohn's disease. Anabolic effects of GH are mediated via phosphorylation of the signal transducer and activator of transcription (STAT)5b transcription factor. Although GH resistance in other settings has been linked to a defect in janus kinase-STAT signaling, the molecular basis for GH resistance in colitis was not known. We hypothesized that the GH-induced phosphorylation of STAT5b would be impaired in colitis, and that TNFalpha blockade would restore GH signaling. METHODS: Growth, body composition, and molecular regulators of GH signaling were determined in interleukin-10 null mice with chronic colitis and wild-type controls, +/- treatment with an anti TNFalpha antibody. RESULTS: Interleukin-10 null mice exhibited significant alterations in growth, body composition, and feed efficiency. Liver insulin-like growth factor 1 expression was reduced in colitic mice. This was associated with down-regulation of GH receptor (GHR) expression and impaired GH-dependent STAT5b activation. Down-regulation of GHR expression was associated with reduced nuclear abundance and DNA binding of the GHR gene-promoter transactivator, Sp3. TNFalpha down-regulated GHR abundance and prevented GH-induced tyrosine phosphorylation of STAT5 in rat hepatocytes in culture. TNFalpha neutralization up-regulated liver GHR abundance and restored GH activation of STAT5 and serum insulin-like growth factor 1 levels in colitic mice; this preceded improvements in weight gain and disease activity. CONCLUSIONS: GH resistance in experimental colitis is caused by down-regulation of GHR expression, thereby reducing GH-dependent STAT5 activation. TNFalpha blockade restores liver GH signaling and improves anabolic metabolism in this setting. PMID- 15887112 TI - Evidence for repatterning of the gastric fundic epithelium associated with Menetrier's disease and TGFalpha overexpression. AB - BACKGROUND & AIMS: Increase of intramucosal transforming growth factor alpha (TGFalpha) levels in the gastric fundus leads to oxyntic atrophy and massive foveolar hyperplasia in both metallothionein (MT)-TGFalpha mice and patients with Menetrier's disease. We have evaluated the hypothesis that increased levels of TGFalpha in the fundus induces an antral pattern of cell differentiation in fundic glands by studying Pdx1, a transcription factor whose expression normally is confined to the gastric antrum. METHODS: Induction of Pdx1 expression was evaluated in Pdx1(lacZ/+)/MT-TGFalpha bigenic mice treated with zinc. The distribution of Pdx1 in MT-TGFalpha mice and Menetrier's disease patients was evaluated with anti-Pdx1 antibodies. Transcript levels were evaluated by quantitative polymerase chain reaction in mouse and human tissues and AGS cells. RESULTS: In Pdx1(lacZ/+) mice, Pdx1 was expressed in antral mucosal cells including gastrin cells and TFF2-expressing deep glandular mucous cells. Zinc treatment for 2 to 8 weeks in Pdx1(lacZ/+)/MT-TGFalpha transgenic mice resulted in expression of Pdx1 throughout the fundus. No ectopic fundic Pdx1 expression was observed in either H. felis-infected or DMP777-treated mice. In MT-TGFalpha mice, 8 weeks of zinc treatment elicited nuclear Pdx1 staining throughout the fundic mucosa. TGFalpha treatment in AGS cells led to increases in Pdx1 and gastrin messenger RNA expression. Fundic sections from Menetrier's disease patients showed nuclear Pdx1 staining throughout the fundic glands. Treatment of a Menetrier's disease patient with an anti-epidermal growth factor receptor monoclonal antibody reduced fundic expression of both Pdx1 and gastrin. CONCLUSIONS: Overexpression of TGFalpha in MT-TGFalpha mice and Menetrier's disease patients elicits ectopic expression in the fundus of Pdx1, consistent with the phenotype of antralization. PMID- 15887113 TI - The role of leukotriene B4 in Clostridium difficile toxin A-induced ileitis in rats. AB - BACKGROUND & AIMS: Clostridium difficile toxin A is a potent intestinal inflammatory agent that has been shown to act at least partially by neurogenic mechanisms involving activation of the transient receptor potential vanilloid 1 (TRPV1) (capsaicin) receptor. We tested the hypothesis that leukotriene B4 (LTB4) mediates the effects of toxin A via activation of the TRPV1 receptor. METHODS: Isolated rat ileal segments were pretreated with pharmacologic agents before intraluminal injection of toxin A or LTB4. After 3 hours, the treated segments were removed and inflammation was assessed by luminal fluid accumulation, myeloperoxidase activity, and histology. RESULTS: LTB4 caused ileitis similar to that caused by toxin A and antagonism of TRPV1 receptors but not LTB4 receptors inhibited LTB4-induced inflammation. LTB4 also stimulated TRPV1-mediated substance P release and pretreatment with a specific substance P-receptor antagonist blocked LTB4-induced substance P action and ileitis. Inhibition of the LTB4 biosynthetic enzyme 5-lipoxygenase inhibited toxin A-induced increases in ileal LTB4 levels and toxin A- but not LTB4-induced ileitis. CONCLUSIONS: LTB4 mediates the inflammatory effects of toxin A via activation of TRPV1 receptors. PMID- 15887114 TI - Serotonin excites neurons in the human submucous plexus via 5-HT3 receptors. AB - BACKGROUND & AIMS: Serotonin (5-hydroxytryptamine [5-HT]) is a key signaling molecule in the gut. Recently, the neural 5-HT3 receptor received a lot of attention as a possible target in functional bowel diseases. Yet, the 5-HT3 receptor-mediated changes in properties of human enteric neurons is unknown. METHODS: We used a fast imaging technique in combination with the potentiometric dye 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl]vinyl]pyridinium betaine to monitor directly the membrane potential changes in neurons of human submucous plexus from surgical specimens of 21 patients. An Ussing chamber technique was used to study 5-HT3 receptor involvement in chloride secretion. RESULTS: Local microejection of 5-HT directly onto ganglion cells resulted in a transient excitation of enteric neurons characterized by increased spike discharge. This response was mimicked by the 5-HT3 receptor agonist, 2-methyl-5 HT, and blocked by the 5-HT3 receptor antagonist, tropisetron. The proportions of 5-HT-responsive nerve cells per ganglion ranged from 25.5% +/- 18.4% in the duodenum to 54.2% +/- 46.9% in the colon. Interestingly, 2-methyl-5-HT did not evoke chloride secretion in the human intestine but it did in the guinea-pig intestine. Specific 5-HT3A and 5-HT3B receptor subunit immunoreactivity as well as 5-HT3A and 5-HT3B receptor-specific messenger RNA were detected in the tissue samples. Based on co-labeling with the pan-neuronal marker HuC/D we conclude that submucous nerve cells potentially express heteromeric 5-HT3A/B receptors. CONCLUSIONS: We show that 5-HT excited human enteric neurons via 5-HT3 receptors, which may comprise both 5-HT3A and 5-HT3B receptor subunits. PMID- 15887115 TI - A secreted low-molecular-weight protein from Helicobacter pylori induces cell cycle arrest of T cells. AB - BACKGROUND & AIMS: Although Helicobacter pylori is recognized by the human immune system, the bacteria are not eliminated and lead to a chronic inflammation of the gastric mucosa. METHODS: We investigated the interaction of H. pylori with human lymphocytes. T and B lymphocytes were isolated from H. pylori-infected patients and stimulated with anti-CD3/CD28 or interleukin-6. RESULTS: Proliferation of lymphocytes was abolished on co-incubation with different H. pylori strains (1-5 bacteria/cell) or with protein extracts of culture supernatants. Inhibition of proliferation was independent of known virulence factors. The factor is a protein or protein complex with an apparent molecular weight between 30 and 60 kilodaltons, clearly distinct from VacA. Although antigen-specific activation of T cells (as shown by nuclear factor of activated T cells [NFAT]-activation, interferon-gamma production, and CD25 or CD69 up-regulation) remained intact, cell-cycle analysis showed that S-phase entry of T cells was inhibited completely by H. pylori. Consequently, stimulated T cells arrested in the G1 phase of the cell cycle. Western blot analysis showed markedly reduced phosphorylation of the retinoblastoma protein (pRb), suggesting inhibition of G1 cyclin-dependent kinase activity. In line with this, activities of cyclin D3 and cyclin E were down regulated, and levels of the cyclin-dependent kinase inhibitor p27Kip1 were increased. Mouse embryonic fibroblasts deficient in p27 showed a decrease in H. pylori-induced inhibition of cell proliferation, suggesting a central role for p27 in mediating H. pylori-induced G1 arrest. CONCLUSIONS: Induction of cell cycle arrest in lymphocytes may be of major significance for the chronic persistence of bacteria in the human stomach. PMID- 15887116 TI - Mucosal adaptation to enteral nutrients is dependent on the physiologic actions of glucagon-like peptide-2 in mice. AB - BACKGROUND & AIMS: Our understanding of the intestinotropic actions of glucagon like peptide-2 (GLP-2)(1-33) is based on pharmacologic studies involving exogenous administration. However, the physiologic role of GLP-2 in mucosal growth and adaptation to nutritional stimulation remains poorly understood. METHODS: The properties of GLP-2(3-33), a GLP-2(1-33) metabolite, were determined in baby-hamster kidney cells transfected with the mouse GLP-2 receptor complementary DNA and in isolated murine intestinal muscle strips. To investigate the role of endogenous GLP-2(1-33) in gut adaptation, GLP-2(3-33) was administered to mice that were re-fed for 24 hours after 24 hours of fasting, and the small intestine was analyzed. GLP-2(3-33) also was injected into rats for analysis of circulating GLP-2(1-33) levels. RESULTS: GLP-2(3-33) antagonized the actions of GLP-2(1-33) in vitro and ex vivo. Fasting mice exhibited small intestinal atrophy (37% +/- 1% decrease in small intestinal weight, 19% +/- 2% decrease in crypt-villus height, and 99% +/- 35% increase in villus apoptosis, P < .05-.01). Adaptive growth in re-fed mice restored all these parameters, as well as crypt-cell proliferation, to normal control levels (P < .05 vs. fasting); these adaptive changes were prevented partially or completely by co administration of GLP-2(3-33) to refeeding mice (by 32% +/- 19% to 103% +/- 15%, P < .05-.01 vs re-fed mice). Exogenous GLP-2(3-33) did not affect endogenous GLP 2(1-33) levels. CONCLUSIONS: These data show that endogenous GLP-2 regulates the intestinotropic response in re-fed mice through modulation of crypt-cell proliferation and villus apoptosis. GLP-2 is therefore a physiologic regulator of the dynamic adaptation of the gut mucosal epithelium in response to luminal nutrients. PMID- 15887117 TI - Inducible nitric oxide synthase up-regulates Notch-1 in mouse cholangiocytes: implications for carcinogenesis. AB - BACKGROUND & AIMS: Inflammatory mediators and cell fate genes, such as the Notch gene family, both have been implicated in cancer biology. Because cholangiocarcinomas arise in a background of inflammation and express the inflammatory mediator inducible nitric oxide synthase (iNOS), we aimed to determine whether iNOS expression alters Notch expression and signaling. METHODS: Notch receptor and ligand expression in human liver was evaluated by immunohistochemistry. The effect of iNOS and NO on Notch-1 expression was examined in cell lines. RESULTS: Notch-1, but not other Notch receptors, were up regulated by cholangiocytes in primary sclerosing cholangitis and cholangiocarcinoma. The colocalization of Notch-1 and iNOS also was observed in large bile ducts from the hilar region of primary sclerosing cholangitis patients. Notch-1 expression in murine cholangiocytes was iNOS dependent. iNOS expression also facilitated Notch signaling by inducing the nuclear translocation of its intracellular domain and the expression of a transcriptional target, hairy and enhancer of split (Hes)-1. The gamma-secretase inhibitor N-[N-(3,5 Difluorophenacetyl-L-alanyl)-S-phenylglycine]-t-butyl ester, which blocks Notch signaling, enhanced tumor necrosis factor-related apoptosis-inducing ligand induced apoptosis in cholangiocarcinoma cells. CONCLUSIONS: These data implicate a direct link between the inflammatory mediator iNOS and Notch signaling, and have implications for the development and progression of cholangiocarcinoma. PMID- 15887118 TI - Human homologue of maid is a useful marker protein in hepatocarcinogenesis. AB - BACKGROUND & AIMS: Human homologue of maid (HHM) is a helix-loop-helix (HLH) transcriptional regulatory protein that is involved in the hepatic stem cell development and differentiation. We analyzed the potential involvement of HHM in hepatocarcinogenesis. METHODS: We analyzed HHM expression in the choline deficient L-amino acid defined (CDAA) diet model of rat hepatocarcinogenesis and in human adenomatous hyperplasia (AH) and hepatocellular carcinoma (HCC) biopsy samples. We assessed the effects of HHM on cell proliferation. We screened proteins that bind to HHM protein using a yeast 2-hybrid screen. RESULTS: High HHM expression was seen in foci and HCC induced in the rat CDAA diet model. HHM protein was expressed in 23 of 32 AH samples (72%), 19 of 28 well-differentiated HCC samples (68%), and 9 of 18 poorly-moderately differentiated HCC samples (50%). Over-expressed HHM enhanced the S phase. HHM interference RNA significantly inhibited cell proliferation. A yeast 2-hybrid screen identified Jun activation domain-binding protein 1 (Jab1) as a binding partner for HHM. We confirmed HHM and Jab1 binding by immunoprecipitation and immunofluorescent histochemistry. The expression of Jab1 was found in human AH and HCC samples. We found an association between levels of expression of HHM and those of Jab1 in AH and HCC tissues examined (P = .027 by chi2 test). CONCLUSIONS: High-level HHM expression was found from the very early stages of hepatocarcinogenesis, suggesting that HHM may be a useful marker protein to detect. PMID- 15887119 TI - Mice heterozygous for a defect in mitochondrial trifunctional protein develop hepatic steatosis and insulin resistance. AB - BACKGROUND & AIMS: Little is known about the role of mitochondrial beta-oxidation in development of nonalcoholic fatty liver disease (NAFLD). Mitochondrial trifunctional protein (MTP) catalyzes long-chain fatty acid oxidation. Recently, we generated a mouse model for MTP deficiency and reported that homozygous (MTPa /-) mice suffer neonatal death. In this study, we investigated effects of heterozygosity for the MTP defect on hepatic oxidative stress, insulin resistance, and development of NAFLD in mice. METHODS: We evaluated liver histopathology, serum alanine aminotransferase (ALT), glucose, fatty acids, and insulin levels in MTPa+/- and MTPa+/+ littermates. Insulin resistance was evaluated using glucose tolerance test (GTT) and insulin tolerance test (ITT). Liver tissues were used to measure triglyceride and fatty acid content, activity of superoxide dismutases (SOD) and glutathione peroxidase (GPx), glutathione (GSH), and cytochrome P-450 2E1 expression. RESULTS: Aging but not young MTPa+/- mice developed hepatic steatosis with elevated ALT, basal hyperinsulinemia, and increased insulin area under curve (AUC) on GTT compared with MTPa+/+ littermates. In response to insulin challenge, aging MTPa+/- mice had slower rate of glucose disappearance and increased glucose AUC. Significant hepatic steatosis and insulin resistance developed concomitantly in the MTPa+/- mice at 9-10 months of age. Aging MTPa+/- mice had higher antioxidant activity of total SOD and GPx, lower GSH, and increased expression of cytochrome P-450 2E1, consistent with increased hepatic oxidative stress. CONCLUSIONS: Heterozygosity for beta oxidation defects predisposes to NAFLD and insulin resistance in aging mice. Impairment of mitochondrial beta-oxidation may play an important role in pathogenesis of NAFLD. PMID- 15887120 TI - Aging is associated with decreased pancreatic acinar cell regeneration and phosphatidylinositol 3-kinase/Akt activation. AB - BACKGROUND & AIMS: The effects of aging on pancreatic acinar cell proliferation have not been clearly defined. Phosphatidylinositol 3-kinase (PI3K)-mediated phosphorylation of Akt is a critical step for proliferation of various cell types and insulin secretion from pancreatic endocrine cells; however, its role in acinar cell proliferation is not known. The purpose of this study was to (1) delineate the effects of aging on pancreatic regeneration after partial pancreatectomy (Px) and (2) define the involvement of the PI3K/Akt pathway in pancreatic regeneration. METHODS: Following partial Px, pancreatic regeneration and activation of the PI3K pathway were compared in young and aged mice. Activation of the PI3K/Akt pathway was evaluated by Akt phosphorylation (pAkt). The role of the PI3K pathway in pancreatic regeneration after partial Px was assessed by effects of a pharmacologic PI3K inhibitor wortmannin or small interfering RNA (siRNA) to the p85alpha regulatory subunit. To confirm further the critical role of the PI3K/Akt pathway in pancreatic acinar cell proliferation, IGF-1-mediated cell proliferation was determined in cultured acinar cells pretreated with wortmannin or p85alpha siRNA. RESULTS: Pancreatic regeneration and pAkt expression after partial Px were significantly decreased with aging. Treatment with wortmannin or p85alpha siRNA reduced pancreatic regeneration after partial Px. The IGF-1-mediated cell proliferation in vitro was completely blocked by wortmannin or p85alpha siRNA but not by the MEK/ERK inhibitor PD98059. CONCLUSIONS: PI3K/Akt activation plays a critical role in the regeneration of pancreatic acini after resection. Furthermore, pancreatic regeneration is markedly attenuated in the aged pancreas most likely because of decreased PI3K/Akt activation. PMID- 15887121 TI - Cystathionine beta synthase deficiency promotes oxidative stress, fibrosis, and steatosis in mice liver. AB - BACKGROUND & AIMS: Cystathionine beta-synthase (CBS) deficiency causes severe hyperhomocysteinemia, which confers diverse clinical manifestations, notably liver disease. To investigate this aspect of hyperhomocysteinemia, we performed a thorough investigation of liver pathology in CBS-deficient mice, a murine model of severe hyperhomocysteinemia. METHODS: The degree of liver injury and inflammation was assessed by histologic examination, by measurements of products of lipid peroxidation, and by formation of carbonyl groups on protein as a measure for the occurrence of protein oxidation. Analysis of profibrogenic, proinflammatory factors and cell apoptosis was performed by Western blots, real time quantitative reverse-transcription polymerase chain reaction, caspase-3 activity, DNA laddering, and TUNEL assay. RESULTS: Histologic evaluation of liver specimens of 8- to 32-week-old CBS-deficient mice showed that CBS-deficient mice develop inflammation, fibrosis, and hepatic steatosis, concomitant with an enhanced expression of tissue inhibitor of metalloproteinase-1, alpha-smooth muscle actin, pro(alpha)1 collagen type I, transforming growth factor-beta1, and proinflammatory cytokines. Moreover, even if the proapoptotic protein Bax was dominantly expressed and Bcl-2 was down-regulated, caspase-3 was not activated, DNA laddering was not detected, and number of positive TUNEL cells was not increased in liver of CBS-deficient mice compared with wild-type mice. CONCLUSIONS: The results show that hyperhomocysteinemia in liver of CBS-deficient mice promotes oxidative stress, which may cause mitochondrial damage in association with activation of hepatic stellate cells, leading to liver injury. The absence of caspase-3 activation, DNA fragmentation, and TUNEL-positive cells shows that protective signals may counteract apoptotic signals in liver of CBS deficient mice. PMID- 15887122 TI - A novel immunocompetent rat model of HCV infection and hepatitis. AB - BACKGROUND & AIMS: Hepatitis C virus (HCV) infects millions of people worldwide. Therapy is limited, and treatment does not produce a sustained response in the majority of patients. Development of new agents has been hampered by the lack of a convenient animal model. The aim of this study was to determine whether an immunocompetent rat, tolerized and transplanted with a human hepatoma cell line (Huh 7 cells), could be used to sustain an HCV infection. METHODS: Fetal rats were tolerized in utero with 10(5) Huh 7 cells. One day after birth, rats were transplanted with 5 x 10(6) Huh 7 cells and, a week later, inoculated with HCV, genotype 1. RESULTS: In tolerized, transplanted, and HCV-infected rats, Huh 7 cells were found in the liver, and HCV viral replication was detected by the presence of negative strand HCV RNA. HCV levels in serum were measured at 11,000 copies/mL at week 4, peaked at 22,500 copies/mL by week 12. In tolerized, transplanted, inoculated rats, but not controls, serum alanine aminotransferase (ALT) values increased to 60 IU/L by week 4 and reached a peak of approximately 120 IU/L by week 13. Histology showed foci of mononuclear infiltrates in portal and central regions. CONCLUSIONS: HCV-inoculated immunocompetent rats tolerized and transplanted with Huh 7 cells support HCV gene expression, viral replication, and develop biochemical and histologic evidence of hepatitis. PMID- 15887123 TI - Obscure-overt gastrointestinal bleeding. PMID- 15887124 TI - Familial mutations in PMS2 can cause autosomal dominant hereditary nonpolyposis colorectal cancer. AB - BACKGROUND & AIMS: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant disorder caused by familial mutations in some of the genes responsible for DNA mismatch repair. Mutations in the MLH1, MSH2, and MSH6 genes have been documented in this disorder, but there have been limited and conflicting data about the role of another mismatch repair gene, PMS2. It has recently been suggested that mutations in the PMS2 gene do not cause an autosomal dominant disorder. In addition, mutations in each of these 4 mismatch repair genes have been shown to cause an autosomal recessive cancer syndrome in children that is distinct from hereditary nonpolyposis colorectal cancer. We reviewed a cohort of patients for evidence of an autosomal dominant disorder caused by a mutation in the PMS2 gene. METHODS: A cohort of tumor samples from patients with features suggestive of HNPCC was tested for microsatellite instability, the molecular hallmark of HNPCC, and exclusive loss of expression of the PMS2 gene in tumor tissue. RESULTS: A kindred was identified with autosomal dominant HNPCC due to a familial loss-of-function mutation in the PMS2 gene. This is the first description of such a kindred. CONCLUSION: Mutations in the PMS2 gene can, like mutations in other mismatch repair genes, cause both autosomal dominant HNPCC in adults and an autosomal recessive cancer syndrome in children. PMID- 15887125 TI - Hepatic gene expression discriminates responders and nonresponders in treatment of chronic hepatitis C viral infection. AB - BACKGROUND & AIMS: Pegylated interferon (IFN)-alpha plus ribavirin is the most effective treatment of chronic hepatitis C but has unpleasant side effects and high costs. A large proportion of patients do not respond to therapy for reasons that are unclear. We used gene expression profiling to investigate the molecular basis for treatment failure. METHODS: Expression profiling was performed on percutaneous needle liver biopsy specimens taken before therapy. Gene expression levels were compared among 15 nonresponder, 16 responder, and 20 normal liver biopsy specimens. Differential gene expression was confirmed using real-time polymerase chain reaction. RESULTS: We identified 18 genes whose expression differed significantly between all responders and all nonresponders (P < .005). Many of these 18 genes are IFN sensitive and 2 (ISG15/USP18) are linked in a novel IFN-regulatory pathway, suggesting a possible rationale for treatment resistance. Using a number of independent classifier analyses, an 8-gene subset accurately predicted treatment response for 30 of 31 patients. The classifier analyses were applicable to patients with genotype 1 infection and were not correlated with viral load, disease activity, or fibrosis. CONCLUSIONS: Hepatic gene expression profiling identified consistent differences in patients who subsequently fail treatment with pegylated IFN-alpha plus ribavirin: up regulation of a specific set of IFN-responsive genes predicts nonresponse to exogenous therapy. These data may be of use in predicting clinical responses to treatment. PMID- 15887126 TI - The role of prostaglandins and other eicosanoids in the gastrointestinal tract. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are generally prescribed to ameliorate symptoms associated with acute pain and chronic inflammatory diseases such as arthritis. Recent epidemiologic studies and clinical trials indicate that use of NSAIDs and cyclooxygenase (COX)-2 selective inhibitors are associated with a reduced risk of certain malignancies, especially gastrointestinal cancer. The cyclooxygenase enzymes are the best known targets of NSAIDs; this diverse class of compounds blocks conversion of arachidonic acid to prostanoids. Prostaglandins and other eicosanoids derived from COX-1 and COX-2 are involved in a variety of physiologic and pathologic processes in the gastrointestinal tract. Recent efforts to identify the molecular mechanisms by which COX-2-derived prostanoids exert their proneoplastic effects have provided a rationale for the possible use of NSAIDs alone or in a combination with conventional or experimental anticancer agents for the treatment or prevention of gastrointestinal cancers. PMID- 15887127 TI - Our new president-David a. Peura, MD. PMID- 15887128 TI - American Gastroenterological Association medical position statement: Role of the gastroenterologist in the management of esophageal carcinoma. PMID- 15887129 TI - American Gastroenterological Association technical review on the role of the gastroenterologist in the management of esophageal carcinoma. PMID- 15887130 TI - Germline PMS2 mutations: one hit or two? PMID- 15887131 TI - Genetically engineered probiotics: a new twist on an old remedy. PMID- 15887132 TI - The biology of cag in the Helicobacter pylori-human interaction. PMID- 15887133 TI - Combined modality therapy for rectal cancer. PMID- 15887134 TI - How good is stent therapy for esophagorespiratory fistula? PMID- 15887135 TI - No rust to judgment. PMID- 15887136 TI - New therapies for lamivudine-resistant hepatitis B infection. PMID- 15887138 TI - Intestinal electrical stimulation: a possible treatment for morbid obesity? PMID- 15887143 TI - Is somatosensory cortex activated during proximal stomach stimulation and the role of insula in visceral pain. PMID- 15887144 TI - Diagnosis of pancreatic cystic neoplasms: a report of the cooperative pancreatic cyst study. PMID- 15887146 TI - Apoptotic markers in assessing patients with chronic hepatitis C. PMID- 15887148 TI - The analysis of gastric volume measurement by SPECT: gastric structure vs. function. PMID- 15887150 TI - Gastrointestinal cancer imaging: deeper than the eye can see. PMID- 15887151 TI - Advances in Barrett's esophagus and esophageal adenocarcinoma. AB - Despite advances in diagnosis and therapy, esophageal adenocarcinoma remains an aggressive and usually lethal tumor. This review focuses on the epidemiology of esophageal adenocarcinoma and its presumed precursor lesion, Barrett's esophagus; the pathogenesis of the cancer; advances in treatment of adenocarcinoma and Barrett's esophagus; and strategies for cancer prevention. Emphasis is placed on recent literature. Although the absolute number of cases of adenocarcinoma in the United States is still small, the incidence of this cancer has increased dramatically in the last 40 years, and adenocarcinoma is now the predominant form of esophageal cancer in this country. Recent evidence suggests that Barrett's esophagus is more prevalent in asymptomatic individuals than previously appreciated. The pathogenesis of Barrett's esophagus is poorly understood. Given that some subjects will have repeated bouts of severe erosive esophagitis and never develop Barrett's esophagus, host factors must play an important role. The utility of neoadjuvant radiation and chemotherapy in those with adenocarcinoma, although they are widely practiced, is not of clear benefit, and some authorities recommend against it. Ablative therapies, as well as endoscopic mucosal resection, hold promise for those with superficial cancer or high-grade dysplasia. Most series using these modalities feature relatively short follow-up, and longer-term data will be necessary to better describe the effects of these therapies. The value of chemoprevention in subjects with dysplastic Barrett's esophagus by use of cyclooxygenase 2 inhibitors, nonsteroidal anti-inflammatory drugs, or proton pump inhibitors is unknown. Similarly, although endoscopic screening is widely practiced, its value in patients with chronic gastroesophageal reflux disease symptoms is of unproven value, and recommending bodies are divided as to its practice. PMID- 15887152 TI - Helicobacter pylori and gastric cancer: a new paradigm for inflammation associated epithelial cancers. AB - Although gastric cancer has been investigated for centuries, the association with Helicobacter pylori infection has been recognized for only the past few decades. Although the disease has been declining in most industrialized countries, it remains the second most common cause of cancer death worldwide and is, in theory, a largely preventable disease. We have gained many new insights and advances from studies of Helicobacter-infected mouse models. These models corroborate findings in human patients, in whom disease outcome is largely determined by the expression of host proinflammatory cytokines. Studies of the cellular origins of cancer in the Helicobacter-infected mouse model has led to the surprising insight that gastric cancer may originate from circulating bone marrow-derived stem cells (BMDC) and not from resident tissue stem cells as previously believed. It is likely that this new BMDC paradigm of epithelial cancer will prove useful in future investigations of gastrointestinal metaplasia and gastrointestinal cancers associated with chronic inflammation. PMID- 15887153 TI - Helicobacter pylori and MALT lymphoma. PMID- 15887154 TI - Pancreatic cancer: basic and clinical aspects. PMID- 15887155 TI - Surgery for pancreatic cancer: recent controversies and current practice. AB - Pancreatic duct carcinoma remains a common disease with a poor prognosis. More than 30,000 Americans will die of the disease in 2004, making it the fourth leading cause of cancer death. Despite significant advances in the treatment of many other human tumors, the 5-year survival rate for persons diagnosed with pancreatic cancer has not changed in decades and remains <5%. This is due both to the inherently aggressive biology of the disease and to its late diagnosis in most cases. Surgical resection of localized disease remains the only hope for cure of pancreatic cancer. Over the past 2 decades, significant advances in diagnostic imaging, staging, surgical technique, and perioperative care have led to marked improvement in the surgical management of pancreatic cancer patients. Operative mortality rates for pancreaticoduodenectomy are now <5% at major centers, and the average length of hospital stay has been reduced to <2 weeks. Improvements in patient outcome after pancreatic cancer surgery have made possible, for the first time, the design and conduct of large adjuvant therapy studies in pancreatic cancer. Such clinical trials are critical for improving outcomes for pancreatic cancer patients. PMID- 15887156 TI - Treatment for pancreatic cancer: current therapy and continued progress. AB - In the last decade, continued efforts in pancreas cancer research have led to the development of new, more effective therapies. Additionally, progress in understanding the molecular processes underlying the development and progression of this disease provides hope for the development of more effective treatment strategies. Recent clinical trials have provided reason for hope that novel chemotherapy combinations and molecularly targeted agents will lead to improved clinical outcomes for patients with this disease. This article will summarize the data that has led to the current standards of therapy for patients with resectable and advanced pancreatic cancer and review new treatment strategies for this disease. PMID- 15887157 TI - Cholangiocarcinoma. PMID- 15887158 TI - Enteroendocrine tumors other than carcinoid: a review of clinically significant advances. AB - Only relatively recently has there been an increased clinical recognition and characterization of the heterogeneous group of rare gastroenteropancreatic neuroendocrine neoplasms. Most have endocrine function and exhibit varying degrees of malignancy. This review summarizes the derivation of these tumors and the advances in their diagnosis and treatment over the past decade and a half. They are varied in their biological behavior and clinical courses and, depending on their cell type, can produce different hormones causing distinct clinical endocrine syndromes (insulinoma [hypoglycemia], gastrinoma [Zollinger-Ellison syndrome (ZES)], vasoactive intestinal peptideoma [VIPoma], watery diarrhea, hypokalemia-achlorhydria [WDHA], glucagonoma [glucagonoma syndrome], and so forth). In addition to surgery for cure or palliation (by excision and a variety of other cytoreductive techniques), they each are treated with anti-hormonal agents or drugs targeted to each tumor's specific product or its effects. The majority have benefited from the gut hormone-inhibiting action of somatostatin analogs. Because of their usual slow rate of growth it is recommended that, even when they are advanced and incurable, unlike in patients with common and more malignant cancers, patients with neuroendocrine tumors often can be palliated and appear to survive longer when managed with an active approach using sequential multimodality treatment. Advances in these various therapies are reviewed and the beneficial emergence of global self-help patient support groups is noted. PMID- 15887159 TI - Colon cancer screening in 2005: status and challenges. PMID- 15887160 TI - Genetic testing for inherited colon cancer. AB - The genes associated with each of the inherited syndromes of colon cancer have now been identified, and genetic testing is available for diagnosis. These syndromes include familial adenomatous polyposis, hereditary nonpolyposis colorectal cancer, Peutz-Jeghers syndrome, juvenile polyposis syndrome, and, possibly, Cowden's syndrome. Clinical genetic testing approaches have been developed for each of these syndromes and are now a part of accepted clinical care. Disease-causing mutations can be found in the majority of families affected with one of the inherited syndromes, and, most importantly, once a mutation is found in an index case of the family, relatives can be tested for the presence or absence of that mutation with near 100% accuracy. Cancer screening and management in syndrome families is then based on the results of genetic testing. For the physician to order and properly interpret genetic tests, a basic understanding of the types of mutations that lead to inherited disease and the methods for detecting them is vital. These issues will be presented. Additional clinical issues somewhat unique to genetic testing include genetic counseling and informed consent for genetic testing, both of which will also be reviewed. Often the most difficult aspect of genetic testing is deciding which patients and families should undergo the testing. Furthermore, this issue is quite specific for each of the syndromes. Thus, following presentation of general principles of selection for genetic testing, a detailed approach for identifying persons who should undergo testing for each of the individual syndromes will be given, together with relevant descriptions of the syndromes. Finally, the ongoing work to discover new and possibly more common but less penetrant colon cancer susceptibility genes that cause common familial colon cancer will be presented. PMID- 15887161 TI - Current status of gastrointestinal carcinoids. AB - Gastrointestinal (GI) carcinoids are ill-understood, enigmatic malignancies, which, although slow growing compared with adenocarcinomas, can behave aggressively. Carcinoids are classified based on organ site and cell of origin and occur most frequently in the GI (67%) where they are most common in small intestine (25%), appendix (12%), and rectum (14%). Local manifestations--mass, bleeding, obstruction, or perforation--reflect invasion or tumor-induced fibrosis and often result in incidental detection at emergency surgery. Symptoms are protean (flushing, sweating, diarrhea, bronchospasm), usually misdiagnosed, and reflect secretion of diverse amines and peptides. Biochemical diagnosis is established by elevation of plasma chromogranin A (CgA), serotonin, or urinary 5 hydroxyindoleacetic acid (5-HIAA), while topographic localization is by Octreoscan, computerized axial tomography (CAT) scan, or endoscopy/ultrasound. Histological identification is confirmed by CgA and synaptophysin immunohistochemistry. Primary therapy is surgical excision to avert local manifestations and decrease hormone secretion. Hepatic metastases may be amenable to cytoreduction, radiofrequency ablation, embolization alone, or with cytotoxics. Hepatic transplantation may rarely be beneficial. Chemotherapy and radiotherapy have minimal efficacy and substantially decrease quality of life. Intravenously administered receptor-targeted radiolabeled somatostatin analogs are of use in disseminated disease. Local endoscopic excision for gastric (type I and II) and rectal carcinoids may be adequate. Somatostatin analogues provide the most effective symptomatic therapy, although interferon has some utility. Overall 5-year survival for carcinoids of the appendix is 98%, gastric (types I/II) is 81%, rectum is 87%, small intestinal is 60%, colonic carcinoids is 62%, and gastric type III/IV is 33%. PMID- 15887162 TI - Liver transplantation for hepatocellular carcinoma. PMID- 15887179 TI - Gene products with evolutionary functions. AB - It is often tacitly assumed that all gene products serve the needs of life functions of the individual carrying the genome. However, a close look at the formation of genetic variations, which are the drivers of biological evolution, reveals a different view. While a majority of the products of genes, such as housekeeping genes and genes essential for each individual, when exposed to particular life conditions respond to the definition given above, other gene products clearly carry out evolutionary functions at the level of populations. Products of these evolution genes act as generators of genetic variations and/or as modulators of the frequency of genetic variation. This is most readily seen with bacterial populations. Many different mechanisms contribute to the occasional, overall formation of genetic variations. These mechanisms can be grouped into three mechanistically and qualitatively different strategies of generating genetic variations. In addition to the activities of evolution genes, specific properties of matter such as tautomery also contribute to the formation of genetic variations. The views that nature cares actively for biological evolution are documented by evidence taken mainly from microbial genetics. Essential elements of the theory of molecular evolution are discussed, as well as the relevance of this theory for higher organisms and its impact on our worldview. PMID- 15887180 TI - Structural analysis of a highly acetylated protein using a curved-field reflectron mass spectrometer. AB - Matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry (MS/MS) were used to determine the multiple acetylation sites in the histone acetyltransferase (HAT): p300-HAT. Partial cleavage of the peptides containing acetylated lysine residues by trypsin provided a set of nested sequences that enabled us to determine that multiple acetylation occurs on the same molecule. At the same time, cleavages resulting in a terminal unacetylated lysine suggested that not all of these sites are fully modified. Using MS and MS/MS, we were able to characterize both the unmodified and acetylated tryptic peptides covering more than 82% of the protein. PMID- 15887181 TI - Two-stage affinity purification for inducibly phosphorylated membrane proteins. AB - Characterisation of tyrosine phosphorylations induced in immune cells in response to inflammatory stimuli may help elucidate the molecular bases of the diversity of immune responses. We have used anti-phosphotyrosine antibodies in combination with cell surface biotinylation in a two-step affinity purification procedure to recover pervanadate-induced tyrosine phosphorylated proteins from sub-cellular compartments, including the cell surface, of murine T cells and macrophages prior to separation by solution-phase isoelectric focussing and one-dimensional gel electrophoresis and identification by tandem mass spectrometry. PMID- 15887182 TI - Proteomic analysis of cell surface proteins from Clostridium difficile. AB - Clostridium difficile is a bacterium that causes disease of the large intestine, particularly after treatment with antibiotics. The bacterium produces two toxins (A and B) that are responsible for the pathology of the disease. In addition, a number of bacterial virulence factors associated with adhesion to the gut have previously been identified, including the cell wall protein Cwp66, the high molecular weight surface layer protein (HMW-SLP) and the flagella. As the genome sequence predicts many other cell wall associated proteins, we have investigated the diversity of proteins in cell wall extracts, with the aim of identifying further virulence factors. We have used a number of methods to remove the proteins associated with the cell wall of C. difficile. Two of the resulting extracts, obtained using low pH glycine treatment and lysozyme digestion of the cell wall, have been analysed in detail by two-dimensional electrophoresis and mass spectrometry. One hundred and nineteen spots, comprising 49 different proteins, have been identified. The two surface layer proteins (SLPs) are the most abundant proteins, and we have also found components of the flagellum. Interestingly, we have also determined that a number of paralogs of the HMW-SLP are expressed, and these could represent targets for further investigation as virulence factors. PMID- 15887183 TI - Characterization of nitroproteome in neuron-like PC12 cells differentiated with nerve growth factor: identification of two nitration sites in alpha-tubulin. AB - Nitric oxide (NO) is a precursor of reactive nitrating species, peroxynitrite and nitrogen dioxide, which modify proteins to generate oxidized species such as 3 nitrotyrosine that has been used as a hallmark of peroxynitrite-mediated oxidative stress on proteins. In the last few years however, a growing body of evidence indicates that NO also regulates a myriad of physiologic responses by modifying tyrosine residues. Looking for the molecular event triggered by NO in nerve growth factor (NGF)-induced neuronal differentiation, we recently reported that in differentiating PC12 cells, the cytoskeleton becomes the main cellular fraction containing nitrotyrosinated proteins, and alpha-tubulin is the major target. In the present work, we focus on the investigation of the sites of tyrosine nitration in alpha-tubulin purified by two-dimensional gel electrophoresis following anti-alpha-tubulin immunoprecipitation of protein extract from NGF-treated PC12 cells. Using Western blotting and matrix-assisted laser desorption/ionization-time of flight analysis, we show for the first time, both in vivo and in vitro, that nitration can occur on alpha-tubulin at sites other than the C-terminus and we positively identify Tyr 161 and Tyr 357 as two specific amino acids endogenously nitrated. PMID- 15887184 TI - Quantitative analysis of protein expression using amine-specific isobaric tags in Escherichia coli cells expressing rhsA elements. AB - We describe the use of amine-specific isobaric tags for protein expression quantification to study the effect of rhsA element over-expression in Escherichia coli. The use of an isobaric tagging strategy facilitates a shotgun approach to proteomic analysis and enables quantitation of up to four samples in parallel, based on the reporter ion series using tandem mass spectrometry (MS/MS). Using a liquid chromatography matrix-assisted laser desorption/ionization approach, 23,139 MS/MS spectra were collected. Five thousand sixty-three peptides derived from 780 proteins were quantified including several lower abundance proteins, such as transcription factors, DnaB and DnaG. More than 65% of the proteins had at least two high confidence peptide matches per protein (p<0.05). Further, a statistical test based on the Grubb's and Rosner's tests was able to discriminate outlier data. The removal of outlier data had no significant effect on the functional categories of proteins that were represented in the study. PMID- 15887185 TI - Improvements in two-dimensional gel electrophoresis by utilizing a low cost "in house" neutral pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis system. AB - Two-dimensional gel electrophoresis (2-DE) is widely used for initial protein separation in proteomics. Commercial products using neutral pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis ((SDS-PAGE)/(Bis (2-hydroxyethyl) imino-tris (hydroxymethyl) methane-HCl, or Bis-Tris)) have greatly improved this technique, but cost and limited sizes restrict their applications. An "in-house" system is presented, resulting in better resolution, separation, and new spot visualization and improved resolution when compared to Tris-HCl gels. Their utility is demonstrated using albumin-depleted serum samples, rabbit heart left ventricle, and human immunodeficiency virus type 1 (HIV-1). PMID- 15887186 TI - Development of a high-throughput method for preparing human urine for two dimensional electrophoresis. AB - There is an increasing interest in analysing the human urinary proteome in the search for biomarkers. However, despite the ease of its collection, urine is a difficult fluid to analyse by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) because of its dilute protein content and high salt levels. Here, we describe a method for high-throughput processing of urine for 2-D PAGE. Urine is filtered and applied to solid phase extraction columns. After washing, the urinary proteins are eluted and freeze dried. The lyophilised powder can then be resuspended in an appropriate buffer for downstream proteomic analysis. PMID- 15887187 TI - Acrylamide-agarose copolymers: improved resolution of high molecular mass proteins in two-dimensional gel electrophoresis. AB - A method was developed in order to analyse high molecular mass proteins by two dimensional (2-D) electrophoresis using a copolymer of acrylamide and allyl agarose instead of Bis cross-linked polyacrylamide (PA) gels in sodium dodecyl sulphate-electrophoresis. In this work, the matrix composition was optimised to improve the resolution of proteins larger than 200 kDa. The new gel type does not entrap large proteins and protein complexes at the application site. Mechanical properties were investigated through rheological measurements, which suggested the formation of a highly entangled elastomeric soft gel. A high 2-D resolution of proteins, extracted from membranes of red blood cells, was obtained in these gels. An example of tryptic digestion, peptide extraction and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry was reported. The results demonstrate that the new gel is fully compatible with mass spectrometry protein analysis. PMID- 15887188 TI - A quantitative proteomic approach using two-dimensional gel electrophoresis and isotope-coded affinity tag labeling for studying human 20S proteasome heterogeneity. AB - Mammalian proteasomes are macromolecular complexes formed of a catalytic 20S core associated to two regulatory complexes. The 20S core complex consists of four stacked rings of seven alpha or beta subunits. Three beta subunits contain a catalytic site and can be replaced by three interferon gamma-inducible counterparts to form the immunoproteasome. Cells may constitutively possess a mixture of both 20S proteasome types leading to a heterogeneous proteasome population. Purified rat 20S proteasome has been separated in several chromatographic fractions indicating an even higher degree of complexity in 20S proteasome subunit composition. This complexity may arise from the presence of subunit isoforms, as previously detected in purified human erythrocyte 20S proteasome. In this study, we have used a quantitative proteomic approach based on two-dimensional gel electrophoresis and isotope-coded affinity tag (ICAT) labeling to quantify the variations in subunit composition, including subunit isoforms, of 20S proteasomes purified from different cells. The protocol has been adapted to the analysis of low quantities of 20S proteasome complexes. The strategy has then been validated using standard proteins and has been applied to the comparison of 20S proteasomes from erythrocytes and U937 cancer cells. The results obtained show that this approach represents a valuable tool for the study of 20S proteasome heterogeneity. PMID- 15887189 TI - Protein microarrays for antibody profiling: specificity and affinity determination on a chip. AB - Protein microarray technology facilitates the detection and quantification of hundreds of binding reactions in one reaction from a minute amount of sample. Proof-of-concept studies have shown that the set-up of sensitive assay systems based on protein arrays is possible, however, the lack of specific capture reagents limits their use. Therefore, the generation and characterisation of capture molecules is one of the key topics for the development of protein array based systems. Recombinant antibody technologies, such as HuCAL (human combinatorial antibody library; MorphoSys, Munich, Germany), allow the fast generation of highly specific binders to nearly any given target molecule. Although antibody libraries comprise billions of members, it is not the selection process, but the detailed characterisation of the pre-selected monoclonal antibodies that presents the bottleneck for the production of high numbers of specific binders. In order to obtain detailed information on the properties of such antibodies, a microarray-based method has been developed. We show that it is possible to define the specificity of recombinant Fab fragments by protein and peptide microarrays and that antibodies can be classified by binding patterns. Since the assay uses a miniaturised system for the detection of antibody-antigen interactions, the observed binding occurs under ambient analyte conditions as defined by Ekins (J. Pharm. Biomed. Anal. 1989, 7, 155-168). This allows the determination of a relative affinity value for each binding event, and a ranking according to affinity is possible. The new microarray based approach has an extraordinary potential to speed up the screening process for the generation of recombinant antibodies with pre-defined selection criteria, since it is intrinsically a high-throughput technology. PMID- 15887190 TI - Protein sequence tags: a novel solution for comparative proteomics. AB - Comparative proteome profiling using stable isotope peptide labelling and mass spectrometry has emerged as a promising strategy. Here, we show the broad potential of our proprietary protein sequence tag (PST) technology. A special feature of PST is its ability to detect a wide variety of proteins including the pharmaceutically relevant membrane and nuclear proteins. This procedure addresses a similar number of proteins, compared to the multidimensional protein identification technology approach, but offers additionally a quantitative analysis with its recently developed quantitative PST version. PMID- 15887191 TI - Degenerate two-photon absorption spectra in azoaromatic compounds. AB - A systematic study of the degenerate two-photon absorption (2PA) spectra of seven azoaromatic compounds in dimethyl sulfoxide solution is reported, which employed the Z-scan technique with femtosecond laser pulses from the bottom of the azo compound absorption bands up to 1100 nm. The 2PA peaks for pseudostilbene-type azo compounds (Disperse Orange (DO) 3, Disperse Red (DR) 13, DR1, DR19, and DR19 Cl) were observed at twice the peak wavelength of the linear absorption. However, such peaks were not observed for other azo compounds (PAMINO and DIAMINO) because of the symmetry of these molecules. A resonance enhancement of the 2PA cross section was observed for all compounds. The 2PA peak and the nonlinear resonance enhancement behavior could be adjusted with a model based on perturbation theory. Such knowledge can be a guideline to the understanding of the 2PA process in azoaromatic compounds. PMID- 15887192 TI - Microstorms in cellular polymers: a route to soft piezoelectric transducer materials with engineered macroscopic dipoles. AB - Cellular polymers can be internally charged by "microstorms" (silent or partial discharges) within the voids of the polymer foam. The resulting material, which carries positive and negative charges on the internal void surfaces, is called a ferroelectret. Ferroelectrets behave like typical ferroelectrics, hence they provide a novel class of ferroic materials. The soft foams are strongly piezoelectric and can be used, in a wide range of applications, as transducers for interconverting mechanical and electrical signals. Herein, an overview is provided on the preparation of cellular polymers by physical foaming (extrusion, biaxial stretching, and controlled inflation by pressure treatments), on their charging by "microstorms", on their piezo- and pyroelectricity, and on analogies to ferroelectrics. Finally, a survey of selected applications is presented. PMID- 15887193 TI - Energy transfer in single hydrogen-bonded water molecules. AB - We study the structure and dynamics of hydrogen-bonded complexes of H2O/HDO and acetone dissolved in carbon tetrachloride by probing the response of the O-H stretching vibrations with linear mid-infrared spectroscopy and femtosecond mid infrared pump-probe spectroscopy. We find that the hydrogen bonds in these complexes break and reform with a characteristic time scale of approximately 1 ps. These hydrogen-bond dynamics are observed to play an important role in the equilibration of vibrational energy over the two O-H groups of the H2O molecule. For both H2O and HDO, the O-H stretching vibrational excitation relaxes with a time constant of 6.3+/-0.3 ps, and the molecular reorientation has a time constant of 6+/-1 ps. PMID- 15887194 TI - The structure of poly(carbonsuboxide) on the atomic scale: a solid-state NMR study. AB - In this contribution we present a study of the structure of amorphous poly(carbonsuboxide) (C3O2)x by 13C solid-state NMR spectroscopy supported by infrared spectroscopy and chemical analysis. Poly(carbonsuboxide) was obtained by polymerization of carbonsuboxide C3O2, which in turn was synthesized from malonic acid bis(trimethylsilylester). Two different 13C labeling schemes were applied to probe inter- and intramonomeric bonds in the polymer by dipolar solid-state NMR methods and also to allow quantitative 13C MAS NMR spectra. Four types of carbon environments can be distinguished in the NMR spectra. Double-quantum and triple quantum 2D correlation experiments were used to assign the observed peaks using the through-space and through-bond dipolar coupling. In order to obtain distance constraints for the intermonomeric bonds, double-quantum constant-time experiments were performed. In these experiments an additional filter step was applied to suppress contributions from not directly bonded 13C,13C spin pairs. The 13C NMR intensities, chemical shifts, connectivities and distances gave constraints for both the polymerization mechanism and the short-range order of the polymer. The experimental results were complemented by bond lengths predicted by density functional theory methods for several previously suggested models. Based on the presented evidence we can unambiguously exclude models based on gamma-pyronic units and support models based on alpha-pyronic units. The possibility of planar ladder- and bracelet-like alpha-pyronic structures is discussed. PMID- 15887195 TI - Influence of pH on the photochemical and electrochemical reduction of the dinuclear ruthenium complex, [(phen)2Ru(tatpp)Ru(phen)2)Cl4, in water: proton coupled sequential and concerted multi-electron reduction. AB - The dinuclear ruthenium complex [(phen)2Ru(tatpp)Ru(phen)2]4+ (P; in which phen is 1,10-phenanthroline and tatpp is 9,11,20,22-tetraaza tetrapyrido[3,2-a:2'3' c:3'',2''-l:2''',3''']-pentacene) undergoes a photodriven two-electron reduction in aqueous solution, thus storing light energy as chemical potential within its structure. The mechanism of this reduction is strongly influenced by the pH, in that basic conditions favor a sequential process involving two one-electron reductions and neutral or slightly acidic conditions favor a proton-coupled, bielectronic process. In this complex, the central tatpp ligand is the site of electron storage and protonation of the central aza nitrogen atoms in the reduced products is observed as a function of the solution pH. The reduction mechanism and characterization of the rich array of products were determined by using a combination of cyclic and AC voltammetry along with UV-visible reflectance spectroelectrochemistry experiments. Both the reduction and protonation state of P could be followed as a function of pH and potential. From these data, estimates of the various reduced species' pKa values were obtained and the mechanism to form the doubly reduced, doubly protonated complex, [(phen)2Ru(H2tatpp)Ru(phen)2]4+ (H2P) at low pH (< or =7) could be shown to be a two-proton, two-electron process. Importantly, H2P is also formed in the photochemical reaction with sacrificial reducing agents, albeit at reduced yields relative to those at higher pH. PMID- 15887196 TI - Template-directed synthesis of multiply mechanically interlocked molecules under thermodynamic control. AB - The template-directed construction of crown-ether-like macrocycles around secondary dialkylammonium ions (R2NH2+) has been utilized for the expedient (one pot) and high-yielding synthesis of a diverse range of mechanically interlocked molecules. The clipping together of appropriately designed dialdehyde and diamine compounds around R2NH2+-containing dumbbell-shaped components proceeds through the formation, under thermodynamic control, of imine bonds. The reversible nature of this particular reaction confers the benefits of "error-checking" and "proof reading", which one usually associates with supramolecular chemistry and strict self-assembly processes, upon these wholly molecular systems. Furthermore, these dynamic covalent syntheses exploit the efficient templating effects that the R2NH2+ ions exert on the macrocyclization of the matched dialdehyde and diamine fragments, resulting not only in rapid rates of reaction, but also affording near quantitative conversion of starting materials into the desired interlocked products. Once assembled, these "dynamic" interlocked compounds can be "fixed" upon reduction of the reversible imine bonds (by using BH3.THF) to give kinetically stable species, a procedure that can be performed in the same reaction vessel as the inital thermodynamically controlled assembly. Isolation and purification of the mechanically interlocked products formed by using this protocol is relatively facile, as no column chromatography is required. Herein, we present the synthesis and characterization of 1) a [2]rotaxane, 2) a [3]rotaxane, 3) a branched [4]rotaxane, 4) a bis [2]rotaxane, and 5) a novel cyclic [4]rotaxane, demonstrating, in incrementally more complex systems, the efficacy of this one-pot strategy for the construction of interlocked molecules. PMID- 15887197 TI - Aqueous foams stabilized solely by silica nanoparticles. PMID- 15887198 TI - Template synthesis of a coordinated tetracarbene ligand with crown ether topology. PMID- 15887199 TI - Exploring polymorphism: the case of benzene. PMID- 15887200 TI - Tetraphosphacyclopentadienyl and triphosphaallyl ligands in iron complexes. PMID- 15887201 TI - {Sn[Zn4Sn4S17]}6-: a robust open framework based on metal-linked penta supertetrahedral [Zn4Sn4S17]10- clusters with ion-exchange properties. PMID- 15887202 TI - Terminal borylene complexes stabilized by a transition-metal base. PMID- 15887203 TI - Synthesis of cis,cis,cis,cis-[5.5.5.4]-1-azafenestrane with discovery of an unexpected dyotropic rearrangement. PMID- 15887204 TI - An electrochemically driven and electrochemically regenerated NOx trap. PMID- 15887205 TI - Ultrastable rhodamine with cucurbituril. PMID- 15887206 TI - The ball-shaped heteropolytungstates [{Sn(CH3)2(H2O)}24{Sn(CH3)2}12(A XW9O34)12]36-. PMID- 15887207 TI - Recent advances in catalytic intramolecular C-H aminations. PMID- 15887208 TI - Dicopper and disilver complexes of octaphyrin(1.1.1.1.1.1.1.1): reversible hydrolytic cleavage of the pyrrolic ring to a keto-imine. PMID- 15887209 TI - The wheel-shaped Cu20 tungstophosphate [Cu20Cl(OH)24(H2O)12(P8W48O184)]25- ion. PMID- 15887210 TI - Liquid inorganic-organic nanocomposites: novel electrolytes and ferrofluids. PMID- 15887211 TI - Gender differences in genetic damage induced by the tobacco-specific nitrosamine NNK and the influence of the Thr241Met polymorphism in the XRCC3 gene. AB - The rapid increase in adenocarcinoma of the lung and mortality amongst women strongly suggests that gender differences exist in sensitivity to certain tobacco carcinogens. In the current study, we performed the mutagen-sensitivity assay, with the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone (NNK), to test the hypothesis that women are more sensitive to the genotoxic effects of NNK than men. Chromosome aberration (CA) frequencies in peripheral blood lymphocytes (PBLs) from 99 patients were evaluated before and after in vitro exposure to NNK. Because the Thr241Met polymorphism in the DNA repair gene XRCC3 is associated with increased risk of tobacco-related cancers, especially among women, we also tested the hypothesis that individuals who inherit the homozygous variant 241Met allele are more sensitive to the genotoxic effects of NNK. CA frequency was significantly higher 1 hr after NNK treatment in women, compared with men (P = 0.02). When smoking and gender were considered together, a significant interaction was observed. PBLs from female smokers had significantly higher frequencies of NNK-induced CA, compared with female nonsmokers 1 hr after treatment (P = 0.02). We observed no overall effect of the Thr241Met polymorphism on NNK-induced CA in men, women, smokers, or nonsmokers. Overall, our data indicate that women are more sensitive to the genotoxic effects of NNK than men. Because in past years smoking among women has increased, and in view of the close correlation between NNK exposure and adenocarcinoma of the lung, our data provide a plausible explanation for the recent increase in the incidence of this cancer among women. PMID- 15887212 TI - Use of the Comet test and micronucleus assay on human white blood cells for in vitro assessment of genotoxicity induced by different drinking water disinfection protocols. AB - Surface water disinfection can lead to the formation of mutagenic/carcinogenic by products derived from reactions with naturally occurring inorganic compounds. We investigated the feasibility and potential usefulness of an integrated approach to genotoxicity analysis of drinking water. The approach employed the Comet and micronucleus (MN) assays to evaluate the DNA and chromosomal damage produced by water extracts in human blood cells. Surface water samples from Lago Trasimeno (Italy) were collected in different seasons (July 2000, October 2000, February 2001, and June 2001), and samples were disinfected with sodium hypochloride (NaClO), chlorine dioxide (ClO(2)), or peracetic acid (PAA). Extracts of untreated and treated water were incubated with primary human leukocytes. The Comet assay revealed both strong seasonal variations and differences between samples processed by the three disinfection protocols. The three disinfectants increased the genotoxicity of the water collected in July 2000 and October 2000, with PAA producing the greatest amount of DNA damage. Extracts of raw water collected in February 2001 produced so much DNA damage that the relative genotoxic potentials of the three disinfectants could not be evaluated. No increase in MN frequency was detected in any of the samples. The multi-endpoint MN assay indicated, however, that our study samples (especially the sample collected in the February 2001) were cytotoxic. We conclude that this integrated approach to genotoxicity assessment may be useful both for the quality control of raw drinking water and to help compare the potential health risks associated with alternative disinfection processes. PMID- 15887213 TI - In vitro studies of the genotoxicity of ionizing radiation in human G(0) T lymphocytes. AB - In an effort to mimic human in vivo exposures to ionizing irradiation, G(0) phase T lymphocytes from human peripheral blood samples were utilized for in vitro studies of the genotoxic effects of (137)Cs low-LET irradiation and (222)Rn high LET irradiation. Both types of radiation induced mutations in the HPRT gene in a dose-dependent manner, with a mutant frequency (MF) = 4.28 + 1.34x + 7.51x(2) for (137)Cs (R(2) = 0.95) and MF = 4.81 + 0.67x for (222)Rn (R(2) = 0.51). Post (137)Cs irradiation incubation in the presence of cytosine arabinoside, a reversible inhibitor of DNA repair, caused an increase in the MF over irradiation alone, consistent with a misrepair mechanism being involved in the mutagenicity of low-LET irradiation. The spectrum of (137)Cs irradiation-induced mutation displayed an increase in macro-deletions (in particular total gene deletions) and rearrangement events, some of which were further defined by either chromosome painting or direct DNA sequencing. The spectrum of (222)Rn irradiation-induced mutation was characterized by an increase in small alterations, especially multiple single base deletions/substitutions and micro-deletions. These studies define the specific response of human peripheral blood T cells to ionizing irradiation in vitro and form a basis for evaluating the genotoxic effects of human in vivo exposure. PMID- 15887214 TI - Ultrasonic glossectomy--simple and bloodless. AB - BACKGROUND: The aim of the study was the evaluation of the feasibility of glossectomy using an ultrasonic-activated surgical instrument. METHODS: This was a prospective study of 13 consecutive patients who underwent glossectomy (12 partial and one total) for carcinoma of tongue with the use of ultrasonic scissors. RESULTS: All 13 patients had glossectomy, with median blood loss of 0 mL. The glossectomies were done with an ultrasonic dissector alone. None of the glossectomies required diathermy, ligature, plication, or other methods for hemostasis. The lingual artery and veins of all 13 patients were controlled by use of the ultrasonic scissors alone. No operative complications occurred, including bleeding or wound healing problems. CONCLUSIONS: The excellent combination of coagulation and the cutting effect of ultrasonic scissors has made glossectomy a simple and bloodless procedure. It is a recommended surgical technique in our surgical armamentarium. PMID- 15887215 TI - Comparative analysis of the mutagenic activity of oxaliplatin and cisplatin in the Hprt gene of CHO cells. AB - Oxaliplatin is a platinum-derived antitumor drug that is active against cisplatin resistant tumors and has lower overall toxicity than does cisplatin. DNA adduct formation is believed to mediate the cytotoxic activity of both compounds; however, the adducts may also be responsible for mutagenic and secondary tumorigenic activities. In this study, we have compared the mutagenicity of oxaliplatin and cisplatin in the Hprt gene of CHO-K1 cells. Both drugs produced dose-related increases in mutant frequency. For 1-hr treatments, oxaliplatin was less mutagenic than cisplatin at equimolar doses, while similar mutant frequencies were induced at equitoxic doses. Sequencing of mutant Hprt genes indicated that the mutation spectra of both oxaliplatin and cisplatin were significantly different from the spontaneous mutation spectrum (P = 0.014 and P = 0.008, respectively). A significant difference was also observed between the spectra of oxaliplatin- and cisplatin-induced mutations (P = 0.033). Although G:C ->T:A transversion was the most common mutation produced by both compounds, oxaliplatin produced higher frequencies of A:T-->T:A transversion than did cisplatin, most commonly at nucleotide 307, and higher frequencies of small deletions/insertions. Also, cisplatin induced tandem base-pair substitutions, mainly at positions 135/136, and a higher frequency of G:C-->A:T transition than did oxaliplatin. These results provide the first evidence that oxaliplatin is mutagenic and that the profiles of cisplatin- and oxaliplatin-induced mutations display not only similarities but also distinctive features relating to the type and sequence-context preference for mutation. Environ. PMID- 15887216 TI - Expression of ets-1 transcription factor in human head and neck squamous cell carcinoma and effect of histamine on metastatic potential of invasive tumor through the regulation of expression of ets-1 and matrix metalloproteinase-3. AB - BACKGROUND: Ets-1 controls the expression of critical genes involved in matrix remodeling. The matrix metalloproteinase-3 (MMP-3) and urokinase type plasminogen activator (uPA) are typical ets-1 responsive genes. Recent studies have shown an increase in histamine synthesis and content in various human neoplasias. We hypothesized that the increased local histamine overproduction contributed to activation of matrix remodeling through the activation of MMP-3 expression of peritumoral fibroblasts by means of ets-1 regulation in head and neck squamous cell carcinomas (HNSCCs). METHODS: Paraffin-embedded sections of 30 HNSCCs were immunostained for ets-1. The presence of ets-1 and MMP-3 mRNA in tumor samples was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR). To simulate stromal reaction in vitro, cultured human mucosal fibroblast was used. The level of ets-1 and MMP-3 mRNA was compared by use of RT-PCR, as was their protein with flow-cytometry, in the presence or absence of basic fibroblast growth factor (bFGF) (10 ng/mL) and histamine (1 microM). RESULTS: Correlation between ets-1 expression and clinicopathologic background was not significant. In all cases, expression of ets-1 was seen in the stroma. In in vitro study, histamine upregulates production of ets-1 and MMP-3 in cultured fibroblast, and bFGF can stimulate histamine expression in fibroblast. Immunofluorescence staining supported the results of RT-PCR and flow cytometry. CONCLUSIONS: Ets-1 expression in HNSCCs has no prognostic value; however, ets-1 plays an important role in tumor-host interaction. Histamine may accelerate the spread of HNSCC through an ets-1-related mechanism. PMID- 15887217 TI - Endoscopic myotomy of the cricopharyngeal muscle with CO2 laser surgery. AB - BACKGROUND: Cricopharyngeal dysfunction may lead to severe dysphagia and aspiration. Several treatment modalities are available, such as external myotomy of the muscle, dilatation, and local infiltration with botulinum toxin. Recently, endoscopic transmucosal myotomies using a CO2 laser have been described. METHODS: Endoscopic cricopharyngeal myotomy using a CO2 laser was performed in 10 consecutive patients with dysphagia supposedly caused by cricopharyngeal muscle dysfunction without Zenker's diverticulum. All patients received prophylactic antibiotics and a feeding tube. Assessment was composed of clinical observation, a questionnaire, and a physical examination including flexible endoscopy and videofluoroscopy. RESULTS: In all the patients, the procedure was feasible and without complications. Improvement of the complaints occurred in most patients. One patient had recurrent dysphagia and required a second endoscopic procedure. CONCLUSIONS: Endoscopic laser surgery seems to be a safe and effective technique to treat cricopharyngeal dysfunction. PMID- 15887218 TI - Interobserver reliability of computed tomography-derived primary tumor volume measurement in patients with supraglottic carcinoma. AB - BACKGROUND: Prior studies have determined that macroscopic ("gross") tumor volume (GTV), as calculated from pretreatment computer tomography (CT), was capable of predicting local control in squamous cell carcinoma arising in different subsites in the head and neck in patients who were treated with nonsurgical organ preservation therapy. The majority of these studies were single-institution, retrospective investigations. Consequently, there has been concern that GTV measurements may not be reproducible by different readers at different institutions. The objective of the current study was to measure the interobserver reliability for GTV measurements for squamous cell carcinoma of the supraglottic larynx (SGSCCA) performed by different readers at different institutions. METHODS: Eight experienced readers (4 neuroradiologists and 4 radiation oncologists) from different institutions independently measured the pretreatment GTV of 20 patients with SGSCCA. The CT scans were obtained from patients entered into the definitive radiation therapy arm of Radiation Therapy Oncology Group protocol 91-11, who had supraglottic carcinoma and underwent pretreatment CT scans of the neck. Statistical analysis focused on interobserver reliability as measured by the intraclass correlation coefficient. RESULTS: The intraclass correlation coefficient was 0.81 (95% lower confidence bound, 0.71). This value was interpreted as "excellent." CONCLUSIONS: GTV measurements were reliable and reproducible when performed by neuroradiologists and radiation oncologists who were experienced in the interpretation of CT scans of the extracranial head and neck in patients with SGSCCA. The result implied that the correlation between GTV and local control should be reproducible across institutions. PMID- 15887220 TI - A phase II study of bortezomib in the treatment of metastatic malignant melanoma. AB - BACKGROUND: Bortezomib is a proteasome inhibitor with manageable clinical toxicity and laboratory evidence of anti-melanoma activity. Therefore, it was considered for clinical testing in patients with metastatic melanoma. METHODS: Patients with metastatic melanoma and adequate hematologic, renal, and hepatic function were treated with bortezomib (a 1.5-mg/m2 intravenous bolus twice weekly for 2 of every 3 weeks). Eligible patients were age > or = 18 years with an Eastern Cooperative Oncology Group performance status of 0-1. The primary goal of the current study was to evaluate the 18-week disease progression-free survival rate and tolerability of bortezomib in these patients. RESULTS: The current study was intended to treat 45 patients. It was closed at the planned interim analysis due to early evidence of insufficient clinical efficacy. Twenty-seven patients with a median age of 56 years (range, 32-77 years)were accrued. There were no major clinical responses to treatment. Only 6 patients (22%) achieved stable disease. Of these 6 patients, 4 were still stable after 4 cycles of treatment, but were removed from the study due to toxicity. The median time to disease progression was 1.5 months (95% confidence interval [95% CI], 1.4-1.6) with a median overall survival of 14.5 months (95% CI, 9-22). Having failed bortezomib, most patients proceeded to other clinical trials. Twenty-six patients were evaluable for toxicity. One patient was removed from the study for other reasons and could not return for the cycle evaluation and thus was never evaluated. Of the 26 patients, no Grade 4/5 treatment-related toxicities (using the National Cancer Institute Common Toxicity Criteria [version 2.0]) were reported. Eleven patients (42%) had Grade 3 toxicities (believed to be at least possibly related to treatment), including sensory neuropathy, thrombocytopenia, constipation, fatigue, ileus, abdominal pain, and infection without neutropenia. The median number of treatment cycles patients received was two (range, one to six treatment cycles). Two patients (7%) had 1 dose delay and 6 patients (22%) had dose reductions during 1 treatment cycle due to adverse events. CONCLUSIONS: Single agent bortezomib, administered twice weekly x 2 weeks, every 3 weeks at a dose of 1.5 mg/m2, was not found to be effective in the treatment of patients with metastatic melanoma. PMID- 15887221 TI - Exploring the charge space of protein-protein association: a proteomic study. AB - The rate of association of a protein complex is a function of an intrinsic basal rate and of the magnitude of electrostatic steering. In the present study we analyze the contribution of electrostatics towards the association rate of proteins in a database of 68 transient hetero-protein-protein complexes. Our calculations are based on an upgraded version of the computer algorithm PARE, which was shown to successfully predict the impact of mutations on k(on) by calculating the difference in Columbic energy of interaction of a pair of proteins. HyPare (http://bip.weizmann.ac.il/HyPare), automatically calculates the impact of mutations on a per-residue basis for all residues of a protein-protein interaction, achieving a precision similar to that of PARE. Our calculations show that electrostatics play a marginal role (<10 fold) in determining the rate of association for about half of the complexes in the database. Strong electrostatic steering, which results in an increase of over 100-fold in k(on), was calculated for about 25% of the complexes. Applying HyPare to all 68 complexes in the database shows that a small number of residues are hotspots for association. About 40% of the hotspots are calculated to increase the rate of association upon mutation, and thus increase binding affinity. This is a much higher ratio than found for hotspots for dissociation, where the large majority cause weaker binding. About 40% of the hotspots are located outside the physical boundary of the binding site, making them ideal candidates for protein engineering. Our data shows that a majority of protein-protein complexes are not optimized for fast association. Hotspots are not evenly distributed between all types of amino acids. About 75% of all hotspots are of charged residues. This is understandable, as a charge-reverse mutant changes the total charge by 2. The small number of hydrophobic residues that are hotspots upon mutation probably relates to their location and surrounding. For 18 out of the 68 complexes in the database, experimental values of k(on) are available. For these, a basal rate of association was calculated to be in the range of 10(4)M(-1)s(-1) to 10(7)M(-1)s( 1). Some of these rates were verified independently from experimental mutant data. The basal rates were correlated with the size of the proteins and the shape of the interface. PMID- 15887222 TI - Squamous cell carcinoma antigen-immunoglobulin M complexes as novel biomarkers for hepatocellular carcinoma. AB - BACKGROUND: Early detection of hepatocellular carcinoma (HCC), one of the most common and deadly tumors worldwide, still is difficult due to the lack of adequate biomarkers that show high sensitivity and specificity. The authors recently demonstrated that squamous cell carcinoma antigen (SCCA) variants were overexpressed remarkably in all surgically resected HCCs. METHODS: For the current study, the authors assessed the presence of SCCA, as a free form and complexed with immunoglobulins, in serum from patients with HCC, cirrhosis, and chronic hepatitis and from healthy control participants and compared SCCA measurement with the measurement of alpha-fetoprotein (AFP) levels. RESULTS: Circulating immune complexes (ICs) composed by SCCA and immunoglobulin M (IgM) IC (SCCA-IgM IC) were undetectable (< 120 arbitrary units [AU]/mL) in serum from a healthy control population (0 of 73 controls); however, 35 of 50 patients with HCC (70%) were reactive for SCCA-IgM IC independent of etiology (mean +/- standard deviation [SD], 2568.5 +/- 6797.3 AU/mL). No correlation was found with AFP levels, which were elevated significantly in only 21 of 50 patients with HCC (42%). By using an AFP cut-off value of 20 ng/mL, 96% of patients with HCC were positive for at least 1 marker. Among cirrhotic patients, the presence of circulating SCCA-IgM IC was displayed in 13 of 50 patients (26%), but at lower levels compared with the patients who had HCC (mean +/- SD, 147.5 +/- 348.3 AU/mL; P < 0.01; Student t test), whereas 9 of 50 patients with chronic hepatitis (18%) were reactive (mean +/- SD, 39.5 +/- 89.7 AU/mL). No significant presence of free SCCA, free anti-SCCA variants IgG or IgM, or SCCA-IgG IC was found. CONCLUSIONS: The study results indicated that SCCA-IgM ICs represent novel serologic biomarkers, which, alone or in combination with AFP, can increase the sensitivity for diagnosing HCC significantly. PMID- 15887223 TI - Similar long-term results of breast-conservation treatment for Stage I and II invasive lobular carcinoma compared with invasive ductal carcinoma of the breast: The University of Pennsylvania experience. AB - BACKGROUND: The objective of the current study was to determine the long-term results of breast-conservation treatment in women with early-stage, invasive lobular carcinoma of the breast. METHODS: Between 1977 and 1995, 1093 women with Stage I and II invasive ductal carcinoma of the breast and 55 women with invasive lobular carcinoma of the breast underwent lumpectomy, axillary lymph node dissection, and radiation treatment. Overall, 49% of the women received adjuvant systemic therapy (chemotherapy and/or hormones). RESULTS: The median age was 52 years for patients in the invasive ductal group and 54 years for patients in the invasive lobular group. The median follow-up was 8.7 years and 10.2 years for patients in the invasive ductal and invasive lobular groups, respectively. A comparison of patients who had invasive lobular carcinoma with patients who had invasive ductal carcinoma showed no difference in the 10-year actuarial rates of overall survival (85% vs. 79%, respectively; P = 0.73), cause-specific survival (93% vs. 84%, respectively; P = 0.85), or freedom from distant metastases (81% vs. 80%, respectively; P = 0.76). The 10-year rates of local failure were 18% for patients with invasive lobular carcinoma and 12% for patients with invasive ductal carcinoma (P = 0.24), and the 10-year rates of contralateral breast carcinoma development for the 2 groups were 12% and 8%, respectively (P = 0.40). CONCLUSIONS: Breast-conservation treatment yielded similar long-term results for women with early-stage, invasive lobular carcinoma and women with the more prevalent invasive ductal carcinoma. PMID- 15887224 TI - New method for protein secondary structure assignment based on a simple topological descriptor. AB - A simple, five-element descriptor, derived from the Delaunay tessellation of a protein structure in a single point per residue representation, can be assigned to each residue in the protein. The descriptor characterizes main-chain topology and connectivity in the neighborhood of the residue and does not explicitly depend on putative hydrogen bonds or any geometric parameter, including bond length, angles, and areas. Rules based on this descriptor can be used for accurate, robust, and computationally efficient secondary structure assignment that correlates well with the existing methods. PMID- 15887226 TI - A new relaxed state in horse methemoglobin characterized by crystallographic studies. AB - A new relaxed state has been characterized in the crystals of horse methemoglobin grown at neutral pH at low ionic concentration and their low humidity variants. The crystals provide an example for improvement in X-ray diffraction quality with reduced solvent content. Only the classical R state has been so far observed in liganded horse hemoglobin. The state characterized in the present study lies in between the R state and the R2 state characterized earlier in liganded human hemoglobin. The results presented here, along with those of earlier studies, suggest that relaxed and tense hemoglobin can access ensembles of states. PMID- 15887227 TI - Aberrant expression of TRAIL in B chronic lymphocytic leukemia (B-CLL) cells. AB - Analysis of peripheral blood (>85% CD19+/CD5+ B) lymphocytes, obtained from 44 patients affected by B chronic lymphoid leukemia (B-CLL), showed that surface TNF related apoptosis inducing ligand (TRAIL) was expressed in all samples and at higher levels with respect to unfractionated lymphocytes and purified CD19+ B cells, obtained from 15 normal blood donors. Of note, in a subset of B-CLL samples, the addition to B-CLL cultures of a TRAIL-R1-Fc chimera, which binds at high affinity to surface TRAIL, significantly decreased the percentage of viable cells with respect to untreated control B-CLL cells, suggesting that surface TRAIL may play an unexpected role in promoting B-CLL cell survival. In spite of the majority of B-CLL lymphocytes expressed variable surface levels of "death receptors" TRAIL-R1 and TRAIL-R2, the addition in culture of recombinant TRAIL increased (>20% vs. controls) the degree of spontaneous apoptosis in only 11/44 of the B-CLL samples, had no effect in 19/44, while it significantly increased leukemic cell survival in 14/44. Taken together, these findings suggest that an aberrant expression of TRAIL might contribute to the pathogenesis of B-CLL by promoting the survival in a subset of B-CLL cells. PMID- 15887228 TI - Essential versus complex autism: definition of fundamental prognostic subtypes. AB - Heterogeneity within the autism diagnosis obscures the genetic basis of the disorder and impedes our ability to develop effective treatments. We found that by using two readily available tests, autism can be divided into two subgroups, "essential autism" and "complex autism," with different outcomes and recurrence risks. Complex autism consists of individuals in whom there is evidence of some abnormality of early morphogenesis, manifested by either significant dysmorphology or microcephaly. The remainder have "essential autism." From 1995 to 2001, 260 individuals who met DSM-IV criteria for autistic disorder were examined. Five percent (13/260) were microcephalic and 16% (41/260) had significant physical anomalies. Individually, each trait predicted a poorer outcome. Together they define the "complex autism" subgroup, comprising 20% (46/233) of the total autism population. Individuals with complex autism have lower IQs (P=0.006), more seizures (P=0.0008), more abnormal EEGs (46% vs. 30%), more brain abnormalities by MRI (28% vs. 13%). Everyone with an identifiable syndrome was in the complex group. Essential autism defines the more heritable group with higher sib recurrence (4% vs. 0%), more relatives with autism (20% vs. 9%), and higher male to female ratio (6.5:1 vs. 3.2:1). Their outcome was better with higher IQs (P=0.02) and fewer seizures (P=0.0008). They were more apt to develop autism with a regressive onset (43% vs. 23%, P=0.02). Analysis of the features predictive of poor outcome (IQ<55, functionally non-verbal) showed that microcephaly was 100% specific but only 14% sensitive; the presence of physical anomalies was 86% specific and 34% sensitive. The two tests combined yielded 87% specificity, 47% sensitivity, and an odds ratio of 4.8:1 for poor outcome. Separating essential from complex autism should be the first diagnostic step for children with autism spectrum disorders as it allows better prognostication and counseling. Definition of more homogeneous populations should increase power of research analyses. PMID- 15887229 TI - Platelet lysates promote mesenchymal stem cell expansion: a safety substitute for animal serum in cell-based therapy applications. AB - Mesenchymal stem cells (MSCs) are considered as emergent "universal" cells and various tissue repair programs using MSCs are in development. In vitro expansion of MSCs is conventionally achieved in medium containing fetal calf serum (FCS) and is increased by addition of growth factors. However, for widespread clinical applications, contact of MSCs with FCS must be minimized since it is a putative source of prion or virus transmission. Therefore, because platelets are a natural source of growth factors, we sought to investigate in vitro MSC expansion in response to platelet lysates (PL) obtained from platelet-rich plasma. Human MSCs were expanded in FCS (+/-bFGF)- or PL-supplemented medium through a process of subculture. We demonstrated that PL-containing medium is enriched by growth factors (platelet-derived growth factors (PDGFs), basic fibroblast growth factor (bFGF), transforming growth factor (TGF-beta), insulin-like growth factor-1 (IGF 1) ...) and showed that PL is able to promote MSC expansion, to decrease the time required to reach confluence, and to increase CFU-F size, as compared to the FCS medium. Furthermore, we demonstrated that MSCs cultured in the presence of PL maintain their osteogenic, chondrogenic, and adipogenic differentiation properties and retain their immunosuppressive activity. Therefore, we propose that PL may be a powerful and safe substitute for FCS in development of tissue- and cellular-engineered products in clinical settings using MSCs. PMID- 15887230 TI - Inhibitory effects of digoxin and ouabain on aldosterone synthesis in human adrenocortical NCI-H295 cells. AB - The present study was to investigate the effects and action mechanisms of digoxin and ouabain on steroidogenesis in human adrenocortical NCI-H295 cells. Administration of digoxin or ouabain for 24 h decreased the basal and angiotensin II (Ang II)-stimulated release of aldosterone by NCI-H295 cells. The conversions of corticosterone (substrate of cytochrome P450 aldosterone synthase, P450c11AS) to aldosterone or deoxycortisol (substrate of cytochrome P450 11beta-hydroxylase, P450c11beta) to cortisol were reduced by digoxin or ouabain. The basal and 22 hydroxy-cholesterol (a membrane-permeable cholesterol, substrate of cytochrome P450 side-chain cleavage enzyme, P450scc)-stimulated pregnenolone release in mitochondria was inhibited by digoxin or ouabain. Digoxin or ouabain suppressed the basal and Ang II-stimulated protein expression of steroidogenic acute regulatory (StAR) protein and P450scc. Incubation of digoxin or ouabain for 24 h reduced P450c11AS mRNA expression in NCI-H295 cells. Digoxin or ouabain (10(-6) M, 24 h)-treated cells showed a lower resting intracellular Ca2+ concentration ([Ca2+]i) and an attenuated response of [Ca2+]i to Ang II. Since no significant cytotoxicity was observed at 10(-6) M digoxin or ouabain, the digoxin- or ouabain induced decrease of aldosterone or cortisol release was not associated with cytotoxicity. These results demonstrate that digoxin or ouabain inhibits the aldosterone or cortisol release via reduction of P450c11AS or P450c11beta and P450scc activities, inhibition of StAR and P450scc protein expression, suppression of P450c11AS mRNA expression, and attenuation of Ca2+ mobilization in NCI-H295 cells. PMID- 15887231 TI - Plasminogen activator inhibitor type 2 (PAI-2) is present in normal human conjunctiva. AB - The purpose was to characterize plasminogen activator inhibitor type 2 (PAI-2) expression in normal human conjunctiva in vivo and in vitro. PAI-2 antigen was assayed by immunostaining and immunoblotting of extracts from normal human conjunctival epithelial lysates and conditioned media (CM) of cultured human conjunctival keratinocytes. Immunostaining of normal human conjunctival epithelia revealed that PAI-2 was found consistently in the superficial keratinocytes and, in some biopsies, also in the lower keratinocyte layers. In all cases, PAI-2 was concentrated around the cell periphery. In extracts of conjunctival epithelia and cultured conjunctival keratinocytes, PAI-2 had an apparent molecular weight of 45 kDa, consistent with the non-glycosylated form. The majority of PAI-2, approximately 90%, was cell associated, however, a small percentage of PAI-2 was released into the CM in a linear manner with time. PAI-2 in the conditioned medium had a higher molecular weight, consistent with a glycosylated form. Conjunctival PAI-2 was active, as shown by its ability to complex with a target enzyme, urokinase plasminogen activator (uPA). Although PAI-2 was detectable both in monolayer (i.e., relatively undifferentiated) conjunctival keratinocyte cultures as well as in stratified (i.e., more differentiated) cultures, steady state levels of PAI-2 were greater in the latter. PAI-2 is constitutively expressed by normal human conjunctival epithelial cells. The expression of PAI-2 throughout all epithelial layers in some biopsies of conjunctiva in vivo contrasts with the previously established distribution of PAI-2 in corneal epithelia, where it is present exclusively in the most superficial (i.e. most highly differentiated) cells. The role of PAI-2 in either tissue is unclear. However, we speculate that its distinct distribution in conjunctival versus corneal epithelia underscores inherent differences between these tissues, and may reflect specific functions of this proteinase inhibitor in both conjunctival and corneal epithelial cells. PMID- 15887232 TI - Interplay between NFBP and NF-kappaB modulates tat activation of the LTR. AB - Interplay of the HIV-1 regulatory protein, Tat, with several cellular factors plays an important role in transcriptional regulation of the viral promoter, the long terminal repeat (LTR). Special attention has been paid to NF-kappaB, a family of inducible transcription factors, which interact with a specific DNA motif within the LTR. Here, we report on the physical and functional interaction of NFBP, a recently identified protein that interacts with the P65 subunit of NF kappaB, with HIV-1 Tat. NFBP colocalizes with Tat in the nucleus and nucleoli, recognizes the amino acid residues 37 to 48 of Tat, and its interaction is modulated by RNA molecules. The interaction of NFBP with Tat modulates the synergism between Tat and P65 in activating LTR transcription. In the absence of the kappaB-binding sites, NFBP augments the TAR-dependent activation by Tat, yet it interferes with the synergistic effect of P65 and Tat on LTR transcription. PMID- 15887233 TI - Effects of soluble laminin on organelle transport and neurite growth in cultured mouse dorsal root ganglion neurons: difference between primary neurites and branches. AB - Laminin, an extracellular matrix molecule, is known to promote neurite growth. In the present study, the effects of soluble laminin on organelle transport and their relation to neurite growth were investigated in cultured dissociated mouse dorsal root ganglion (DRG) neurons. Laminin added into the extracellular medium was deposited on the surface of DRG neurons. DRG neurons incubated with soluble laminin exhibited branched, long, and thin neurites. Time-lapse study demonstrated that many small-diameter branches were newly formed after the addition of laminin. Thus, the growths of large-diameter primary neuritis, arising from cell bodies and branches extended from growth cones of primary neuritis, were analyzed separately. Laminin decreased the growth rate of primary neurites but increased that of branches. In primary neurites, acute addition of laminin rapidly decreased organelle movement in the neurite shaft and growth cone, accompanied by slowing of the growth cone advance. Branching of primary neurites occurred in response to laminin in some growth cones. In these growth cones, organelles protruded into nascent branches. In branches, soluble laminin increased organelle movement in the growth cone and the distal portion of the shaft. These results suggest that laminin inhibits the elongation of primary neurites but promotes branching and elongation of branches, all of which seem to be closely related to organelle transport. PMID- 15887234 TI - Novel glycosaminoglycan mimetic (RGTA, RGD120) contributes to enhance skeletal muscle satellite cell fusion by increasing intracellular Ca2+ and calpain activity. AB - Glycosaminoglycans (GAG) are classes of molecules that play an important role in cellular processes. The use of GAG mimetics called regenerating agent (RGTA) represents a tool to investigate the effect of GAG moiety on cellular behavior. A first member of the RGTA family (RG1192), a dextran polymers with defined amounts of sulfate, carboxymethyl, as well as hydrophobic groups (benzylamide), was shown to stimulate skeletal muscle repair after damage and myoblast differentiation. To obtain a comprehensive insight into the mechanism of action of GAG mimetics, we investigated the effect on myoblast differentiation of a novel RGTA, named RGD120, which was devoid of hydrophobic substitution and had ionic charge similar to heparin. Myoblasts isolated from adult rat skeletal muscles and grown in primary cultures were used in this study. We found that chronic treatment with RGD120 increased the growth of adult myoblasts and induced their precocious fusion into myotubes in vitro. It also partially overcame the inhibitory effect of the calpain inhibitor N-acetyl-leu-leu-norleucinal (ALLN) on these events. Western blot and zymography analyses revealed that milli calpain was slightly increased by RGD120 chronic treatment. In addition, using fluorescent probes (Indo-1 and Boc-leu-met-MAC), we demonstrated that RGD120 added to prefusing myoblast cultures accelerates myoblast fusion into myotubes, induced an increase of cytosolic free calcium concentration, and concomitantly an increase of intracellular calpain protease activity. Altogether, these results suggested that the efficiency of RGD120 in stimulating myogenesis might be in part explained through its effect on calcium mobilization as well as on the calpain amount and activity. PMID- 15887235 TI - Developmental control via GATA factor interplay at chromatin domains. AB - Despite the extraordinary task of packaging mammalian DNA within the constraints of a cell nucleus, individual genes assemble into cell type-specific chromatin structures with high fidelity. This chromatin architecture is a crucial determinant of gene expression signatures that distinguish specific cell types. Whereas extensive progress has been made on defining biochemical and molecular mechanisms of chromatin modification and remodeling, many questions remain unanswered about how cell type-specific chromatin domains assemble and are regulated. This mini-review will discuss emerging studies on how interplay among members of the GATA family of transcription factors establishes and regulates chromatin domains. Dissecting mechanisms underlying the function of hematopoietic GATA factors has revealed fundamental insights into the control of blood cell development from hematopoietic stem cells and the etiology of pathological states in which hematopoiesis is perturbed. PMID- 15887236 TI - HER-2: a biomarker at the crossroads of breast cancer immunotherapy and molecular medicine. AB - The oncoprotein encoded by the HER-2 oncogene is a member of the HER family of receptor tyrosine kinases and is actually the first successfully exploited target molecule in new biomolecular therapies of solid tumors. The association of HER-2 overexpression with human tumors, its extracellular accessibility, as well as its involvement in tumor aggressiveness are all factors that make this receptor an appropriate target for tumor-specific therapy. In addition, HER-2 overexpression fosters its immunogenicity, as shown by the frequency of B and T cell-mediated responses against this oncoprotein in cancer patients, and it is being investigated as a promising molecule for either passive and active immunotherapy strategies. This review summarizes a number of immune intervention approaches that target HER-2 in breast cancer. PMID- 15887237 TI - Arachidonic acid mediates non-capacitative calcium entry evoked by CB1 cannabinoid receptor activation in DDT1 MF-2 smooth muscle cells. AB - Cannabinoid CB1-receptor stimulation in DDT1 MF-2 smooth muscle cells induces a rise in [Ca2+]i, which is dependent on extracellular Ca2+ and modulated by thapsigargin-sensitive stores, suggesting capacitative Ca2+ entry (CCE), and by MAP kinase. Non-capacitative Ca2+ entry (NCCE) stimulated by arachidonic acid (AA) partly mediates histamine H1-receptor-evoked increases in [Ca2+]i in DDT1 MF 2 cells. In the current study, both Ca2+ entry mechanisms and a possible link between MAP kinase activation and increasing [Ca2+]i were investigated. In the whole-cell patch clamp configuration, the CB-receptor agonist CP 55, 940 evoked a transient, Ca2+-dependent K+ current, which was not blocked by the inhibitors of CCE, 2-APB, and SKF 96365. AA, but not its metabolites, evoked a transient outward current and inhibited the response to CP 55,940 in a concentration dependent manner. CP 55,940 induced a concentration-dependent release of AA, which was inhibited by the CB1 antagonist SR 141716. The non-selective Ca2+ channel blockers La3+ and Gd3+ inhibited the CP 55,940-induced current at concentrations that had no effect on thapsigargin-evoked CCE. La3+ also inhibited the AA-induced current. CP 55,940-induced AA release was abolished by Gd3+ and by phospholipase A2 inhibition using quinacrine; this compound also inhibited the outward current. The CP 55,940-induced AA release was strongly reduced by the MAP kinase inhibitor PD 98059. The data suggest that in DDT1 MF-2 cells, AA is an integral component of the CB1 receptor signaling pathway, upstream of NCCE and, via PLA2, downstream of MAP kinase. PMID- 15887238 TI - Induction of heparin-binding EGF-like growth factor and activation of EGF receptor in imatinib mesylate-treated squamous carcinoma cells. AB - Imatinib mesylate is a tyrosine kinase inhibitor of the ABL, platelet-derived growth factor receptor (PDGFR), and c-kit kinases. Inhibition of BCR-ABL and c kit accounts for its clinical activity in leukemia and sarcoma, respectively. In this report, we describe other cellular targets for imatinib. Treatment of head and neck squamous carcinoma cells with clinically relevant concentrations of imatinib-induced changes in cell morphology and growth similar to changes associated with epidermal growth factor receptor (EGFR) activation. Imatinib induced changes were blocked with the EGFR antagonist cetuximab, which suggested direct involvement of EGFR in this process. Western blot analysis of cells incubated with imatinib demonstrated activation of EGFR and downstream signaling that was reduced by inhibition of mitogen-activated protein/extracellular signal regulated kinase kinase 1 (MEK1) and EGFR, but not Her2/ErbB2. An in vitro kinase assay showed that imatinib did not directly affect EGFR kinase activity, suggesting involvement of EGFR-activating molecules. Inhibitors and neutralizing antibodies against heparin-binding epidermal growth factor-like growth factor (HB EGF), and to a lesser extent transforming growth factor-alpha, reduced imatinib mediated mitogen activated protein kinase (MAPK) activation. Imatinib stimulated the rapid release of soluble HB-EGF and the subsequent induction of membrane bound HB-EGF, which correlated with biphasic MAPK activation. Together, these results suggested that imatinib affects EGFR activation and signaling pathways through rapid release and increased expression of endogenous EGFR-activating ligands. Although, imatinib primarily inhibits tyrosine kinases, it also stimulates the activity of EGFR tyrosine kinase in head and neck squamous tumors. This finding demonstrates the need for careful use of this drug in cancer patients. PMID- 15887239 TI - Vitamin C transiently arrests cancer cell cycle progression in S phase and G2/M boundary by modulating the kinetics of activation and the subcellular localization of Cdc25C phosphatase. AB - Regulation of cell cycle progression involves redox (oxidation-reduction) dependent modification of proteins including the mitosis-inducing phosphatase Cdc25C. The role of vitamin C (ascorbic acid, ASC), a known modulator of the cellular redox status, in regulating mitotic entry was investigated in this study. We demonstrated that vitamin C inhibits DNA synthesis in HeLa cells and, mainly the form of dehydroascorbic acid (DHA), delays the entry of p53-deficient synchronized HeLa and T98G cancer cells into mitosis. High concentrations of Vitamin C caused transient S and G2 arrest in both cell lines by delaying the activation of the M-phase promoting factor (MPF), Cdc2/cyclin-B complex. Although vitamin C did not inhibit the accumulation of cyclin-B1, it may have increased the level of Cdc2 inhibitory phosphorylation. This was achieved by transiently maintaining Cdc25C, the activator of Cdc2, both in low levels and in a phosphorylated on Ser216 inactive form that binds to 14-3-3 proteins contributing thus to the nuclear exclusion of Cdc25C. As expected, vitamin C prevented the nuclear accumulation of Cdc25C in both cell lines. In conclusion, it seems that vitamin C induces transient cell cycle arrest, at least in part, by delaying the accumulation and the activation of Cdc25C. PMID- 15887240 TI - Thermotolerance induced at a mild temperature of 40 degrees C protects cells against heat shock-induced apoptosis. AB - Apoptosis constitutes a response of organisms to various physiological or pathological stimuli, and to different stresses. The ability of thermotolerance induced at a mild temperature of 40 degrees C to protect against activation of the apoptotic cascade by heat shock was investigated. When Chinese hamster ovary and human adenocarcinoma cervical cells were pretreated at 40 degrees C for 3 h, they were resistant to subsequent lethal heat shock at 43 degrees C. Induction of thermotolerance at 40 degrees C led to increased expression of heat shock proteins 27, 32, 72, and 90. Heat shock induced apoptotic events at the mitochondrial level, involving a decrease in membrane potential, translocation of Bax to mitochondria, and liberation of cytochrome c into the cytosol. These events were diminished in thermotolerant cells. Heat shock (42-45 degrees C) caused activation of initiator caspase-9 and effector caspases-3, -6, and -7, relative to controls at 37 degrees C. Activation of caspases was decreased in thermotolerant cells. Heat shock caused fragmentation of the caspase substrate, inhibitor of caspase-activated DNase. Fragmentation was diminished in thermotolerant cells. Thermotolerance afforded protection against heat shock induced nuclear chromatin condensation, but not against necrosis. PMID- 15887241 TI - Stomodeal and neurohypophysial placodes in Ciona intestinalis: insights into the origin of the pituitary gland. AB - The ascidian larva has a central nervous system which shares basic characteristics with craniates, such as tripartite organisation and many developmental genes. One difference, at metamorphosis, is that this chordate-like nervous system regresses and the adult's neural complex, composed of the cerebral ganglion and associated neural gland, forms. It is known that neural complex differentiation involves two ectodermal structures, the neurohypophysial duct, derived from the embryonic neural tube, and the stomodeum, i.e. the rudiment of the oral siphon; nevertheless, their precise role remains to be clarified. We have shown that in Ciona intestinalis, the neural complex primordium is the neurohypophysial duct, which in the early larva is a short tube, blind anteriorly, with its lumen in continuity with that of the central nervous system, i.e. the sensory vesicle. The tube grows forwards and fuses with the posterior wall of the stomodeum, a dorsal ectodermal invagination of the larva. The duct then loses posterior communication with the sensory vesicle and begins to grow on the roof of the vesicle itself. The neurohypophysial duct differentiates into the neural gland rudiment; its dorsal wall begins to proliferate neuroblasts, which migrate and converge to build up the cerebral ganglion. The most anterior part of the neural gland organizes into the ciliated duct and funnel, whereas the most posterior part elongates and gives rise to the dorsal strand. The hypothesis that the neurohypophysial duct/stomodeum complex possesses cell populations homologous to the craniate olfactory and adenohypophysial placodes and hypothalamus is discussed. PMID- 15887242 TI - Early post-larval development of the endoparasitic platyhelminth Mesocestoides corti: trypsin provokes reversible tegumental damage leading to serum-induced cell proliferation and growth. AB - Mesocestoides corti is a suitable in vitro model for studying the development of human endoparasitic platyhelminthes. Treatment with trypsin, supplemented with fetal bovine serum (FBS), induces M. corti development from larvae (tetrathyridia) to segmented adult worm; however, the role of this protease and of FBS in post-larval development induction remains unknown. To characterize the participation of trypsin enzymatic activity and of FBS in the induction of tetrathyridia growth and development, both stimuli were added to the larvae either together or sequentially. Additionally, specific inhibition of trypsin activity was also monitored. Finally, the effect of the enzyme on the parasite tegument as well as the proliferative activity and location of proliferating cells after induction of tetrathyridia development were also studied. We conclude that trypsin-induced tetrathyridia development to adult worm is FBS-dependent and that the effect of serum factors is dependent upon a previous trypsin-induced reversible damage to the larva tegument. In dividing and non-dividing tetrathyridia, proliferative activity of cells is mainly located within the apical massif in the anterior region and nerve cords of larvae, respectively. In tetrathyridia stimulated to develop to adult worms, an intense proliferative activity is evident along the nerve cords. Our results suggest that in natural infections the tetrathyridia tegument is temporally made permeable to growth factors by proteolytic enzyme activity in the intestine juice of the definitive host, thus leading to development to adult worms. PMID- 15887243 TI - ETV6 gene rearrangements in invasive breast carcinoma. AB - The ETV6/TEL gene encodes a transcription factor frequently rearranged in several types of cancer. We looked for ETV6 rearrangements in invasive breast cancer using fluorescence in situ hybridization (FISH) of BAC probes on sections of tissue microarrays containing 632 tumor samples. Of these samples, signal of sufficient quality for screening by FISH was obtained for 356. Five cases (one lobular, one nontypical secretory, one mixed, and two ductal carcinomas) showed ETV6 rearrangement. PMID- 15887244 TI - Evolvability of cell specification mechanisms. AB - The architecture of gene action during development is relevant to phenotypic evolution as it links genotype to morphological phenotype. Analysis of development at the level of cell fate specification mechanisms illuminates some of the properties of developmental evolution. In this article, we first review examples of evolutionary change in mechanisms of cell fate specification, with an emphasis on evolution in the dependence on inductive signaling and on evolution of the mechanisms that result in spatial asymmetries. We then focus on properties of development that bias possible phenotypic change and present how the distribution of phenotypes that are available by mutational change of the starting genotype can be experimentally tested by systematic mutagenesis. We finally discuss ways in which selection pressures on phenotypes can be inferred from a comparison of the phenotypic spectrum found on mutation with that found in the wild. PMID- 15887245 TI - Curcumin prevents methylglyoxal-induced oxidative stress and apoptosis in mouse embryonic stem cells and blastocysts. AB - Methylglyoxal (MG) is a reactive dicarbonyl compound endogenously produced mainly from glycolytic intermediates. Elevated MG levels in diabetes patients are believed to contribute to diabetic complications. MG is cytotoxic through induction of apoptosis. Curcumin, the yellow pigment of Curcuma longa, is known to have antioxidant and anti-inflammatory properties. In the present study, we examined the effect of curcumin on apoptotic biochemical events caused by incubation of ESC-B5 cells with MG. Curcumin inhibited the MG-induced DNA fragmentation, caspase-3 activation, cleavage of PARP, mitochondrial cytochrome c release, and JNK activation. Importantly, curcumin also inhibited the MG stimulated increase of reactive oxygen species (ROS) in these cells. In addition, we demonstrated that curcumin prevented the MG-induced apoptosis of mouse blastocysts isolated from pregnant mice. Moreover, curcumin significantly reduced the MG-mediated impairment of blastocyst development from mouse morulas. The results support the hypothesis that curcumin inhibits MG-induced apoptosis in mouse ESC-B5 cells and blastocysts by blocking ROS formation and subsequent apoptotic biochemical events. PMID- 15887246 TI - Contribution of BRCA1 and BRCA2 germ-line mutations to the incidence of breast cancer in young women: results from a prospective population-based study in France. AB - The prevalence of BRCA1/2 germ-line mutations was assessed in a prospective population-based series of early-onset breast cancer (BC) patients in France, and the usefulness of a clinical assessment of hereditary BC risk, based on multiple criteria including pedigree structure, was evaluated. Through the Rhone region BC registry, 232 women diagnosed with BC before 46 years of age were included. They were tested for BRCA1/2 mutations an average of 10 months after diagnosis. All the women were classified according to their family history of cancer: high risk of hereditary breast cancer (HBC), low risk of HBC, isolated BC, and unknown HBC risk. Deleterious mutations were observed in 21 women (9.1%): 15 (6.5%) BRCA1 and 6 (2.6%) BRCA2. Mutations were more prevalent in women who developed BC before age 41 than in women who developed BC between ages 41 and 45 (12.8% versus 5.2%, respectively, P = 0.04). A high prevalence of BRCA1/2 mutations was found among women in the high-risk category with particular family features (i.e., small family size, predominantly male pedigree, specific cancers; 23.5%) and among women with isolated BC before age 41 and with five or fewer close adult female relatives (16.6%). According to the 10% probability level recommended by the American Society of Clinical Oncology guidelines for genetic testing of cancer, BRCA1/2 mutation screening should be considered for all women diagnosed before age 41, except for those with isolated BC in a large pedigree including multiple unaffected female relatives. The clinical assessment of HBC risk that we have developed should help in the decision to perform genetic testing. PMID- 15887247 TI - BRG1 loss in MiaPaCa2 cells induces an altered cellular morphology and disruption in the organization of the actin cytoskeleton. AB - BRG1 and Brahma are critical and mutually exclusive subunits of the multi constituent SWI/SNF chromatin remodeling complexes. These complexes play a key role in transcriptional regulation by dynamically altering chromatin architecture. Although the two proteins are very similar in structure, murine models demonstrate a clear dichotomy in BRG1/BRM function as heterozygous loss of BRG1 results in tumor development whereas homozygous loss of BRM does not. BRG1 and/or BRM protein is absent or disrupted in approximately 17% of all human adenocarcinomas. Concomitant loss is frequent in non-small cell lung carcinomas and incurs a negative prognosis. The mechanism(s) whereby loss of BRG1 (but apparently not BRM) may contribute to tumor development and/or progression is/are ill defined. In this study, we employ MiaPaCa2, a human pancreatic adenocarcinoma cell line that lacks BRM but retains BRG1 expression to evaluate the impact of BRG1 and BRM individually on growth and tumorigenicity. We show that the MiaPaca2 cell line can apparently tolerate only very low levels of BRM after restoration of stable expression. Reduction of expression of BRG1 via shRNAi in stable clones of MiaPaCa2 results in a marked change in morphology and alterations in actin cytoskeletal organization but does not appear to exert a significant effect on in vitro growth of the cell line. Our results implicate a role for the SWI/SNF complex in the regulation of cellular differentiation. PMID- 15887248 TI - Androgen receptor regulates Cdc6 in synchronized LNCaP cells progressing from G1 to S phase. AB - We have shown previously that androgen receptor (AR) activity is required for the progression of cells from G(1) to S phase. In an attempt to elucidate the mechanism of androgen- and androgen-receptor-mediated proliferation of prostate cancer cells, we studied the effect of anti-androgen bicalutamide (Casodex) on the expression of cell-cycle regulatory genes in synchronized LNCaP cells progressing from G(1) to S phase. LNCaP cells were synchronized by isoleucine deprivation. Expression of cell-cycle regulatory genes in S phase control cells versus Casodex-treated cells that fail to enter S phase was studied using a microarray containing cDNA probes for 111 cell-cycle specific genes. RT-PCR and Western-blots were used to validate microarray data. Casodex blocked synchronized LNCaP cells from entering S phase. Microarrays revealed downregulation of eight genes in cells prevented from entering into S phase by Casodex. Of these eight genes, only Cdc6, cyclin A, and cyclin B were downregulated at both the mRNA and protein level in Casodex treated cells as compared to control cells. The mRNA and protein levels of Cdc6 increased as synchronized LNCaP cells progressed from G(1) to S phase, and were attenuated in Casodex-treated cells failed to enter S phase. Cyclins A and B were detected when cells entered S phase, but not when they were in G(1) phase. Like Cdc6, the levels of both cyclins A and B were attenuated in Casodex-treated cells. AR may play an important role in the onset of DNA synthesis in prostate cancer cells by regulating the expression and stability of Cdc6, which is critically required for the assembly of the pre-replication complex(pre-RC). PMID- 15887249 TI - Akt-induced promotion of cell-cycle progression at G2/M phase involves upregulation of NF-Y binding activity in PC12 cells. AB - Akt is a key downstream effector of the PI3K signaling pathway and plays a role in cell growth and survival. Expression of a myristoylated constitutively active form of Akt (myr-Akt) in PC12 cells could override cell-growth arrest at G2/M phase and apoptosis that were induced by etoposide treatment. On the other hand, inactivation of Akt by expression of its dominant negative mutant form (km-Akt) inhibited cell proliferation by arresting the cells at G2/M phase. Expression of myr-Akt also led to an increase in the protein and mRNA levels of CDK1 and cyclin B1. Furthermore, EMSA data revealed that expression of myr-Akt promoted the binding of NF-Y to the consensus CCAAT promoter sequence, whereas expression of km-Akt almost completely abolished it. Moreover, the Akt activity was minimal in the cells that were arrested at G2/M phase by nocodazole treatment, but reached to a maximal level as the cells progressed to mitosis and G1 phase upon removal of the drug. Treatment with Akt inhibitors, but not with those of MEK or p70S6K, blocked the release of the cells from the nocodazole-induced G2/M arrest, further revealing that the Akt activity is required for G2/M phase transition. These results suggest that Akt facilitate cell-cycle progression at G2/M phase in PC12 cells and this Akt activity is correlated with upregulation of NF-Y DNA-binding activity and cyclin B1/CDK1 gene expression. PMID- 15887250 TI - Atypical PKC-zeta and PKC-iota mediate opposing effects on MCF-7 Na+/K+ATPase activity. AB - We demonstrated previously that in serum-starved MCF-7 breast cancer cell line, Ang II increased Na+/K+ATPase activity and activated the protein kinase C zeta (PKC-zeta) (Muscella et al., 2002 J Endocrinol 173:315-323; 2003 J Cell Physiol 197:61-68.). The aim of the present study was to investigate the modulation of the activity of the Na+/K+ATPase by PKC-zeta in MCF-7 cells. Here, using serum starved MCF-7 cells, we have demonstrated that the effect of Ang II on the Na+/K+ATPase activity was inhibited by a synthetic myristoylated peptide with sequences based on the endogenous PKC-zeta pseudosubstrate region (zeta-PS) and by high doses of GF109203X, inhibitor of PKCs. When MCF-7 cells, grown in 10% fetal bovine serum (FBS), were stimulated with Ang II a dose- and time-dependent inhibition of the Na+/K+ATPase activity was obtained. Under this growth condition we found that mRNAs for AT1, AT2, and for Na+/K+ATPase alpha1 and alpha3 subunits were unchanged; besides both the activity of the Na+/K+ATPase and the level of PKC-zeta also were unaffected by the serum. The atypical PKC-iota level (present in very low abundance in serum-starved MCF-7) was increased and Ang II provoked its translocation from the cytosol to plasma membrane. PKC-zeta was localized to the membrane, and upon Ang II treatment its cellular localization did not change. The Ang II-mediated decrease of the Na+/K+ATPase activity was inhibited by high doses of GF109203X but not by zeta-PS, thus indicating that such effect was not due to PKC-zeta activity. The treatment of cells with PKC-iota antisense oligodeoxynucleotides inhibited the effects of Ang II on the Na+/K+ATPase activity. Additionally, the effect of Ang II on Na+/K+ATPase activity was also blocked by the phosphatidylinositol 3-kinase (PI3K) inhibitors, wortmannin and LY294002, and by the actin depolymerizing agents, cytochalasin D. In conclusion, in MCF-7 cells Ang II modulates the Na+/K+ATPase activity by both atypical PKC zeta/-iota. The effects of Ang II are opposite depending upon the presence of the serum-sensitive PKC-iota, with the inhibitory effect possibly due to the redistribution of sodium pump from plasma membrane to the inactive intracellular pool. PMID- 15887251 TI - Perception of local DC and AC electric fields in humans. AB - The goal of this study was to address some of the factors that contribute to the human ability to detect the presence of weak electric fields generated by direct current (DC) and alternating current (AC) sources. An exposure chamber allowed us to expose a limited surface of the body (forearm and hand) to DC fields of up to 65 kV/m and AC fields up to a maximum of 35 kV/m (frequency 60 Hz). Perception was examined using a staircase procedure and a rating procedure derived from signal detection theory. Sixteen subjects participated in the experiments, and none detected the local DC fields. In contrast, 9/16 subjects were sensitive to local AC electric fields, although detection thresholds (index of sensitivity, d' = 1.0) were widely variable between subjects. When regional exposure was limited to the dorsal forearm, performance was similar to that seen when the forearm and hand were exposed. In contrast, subjects did not reliably detect the AC electric fields when exposure was limited to the hand (either hairy or glabrous skin), although a minority of subjects (3/9) showed some evidence of detecting fields presented to the glabrous palm. Subjects were unable to detect AC electric fields when the hair was removed from the forearm and hand, suggesting that the evoked sensation is mainly dependent on movement of hair located in the exposed region. PMID- 15887252 TI - Effects of GSM signals on auditory evoked responses. AB - The article presents a study of the influence of radio frequency (RF) fields emitted by mobile phones on human cerebral activity. Our work was based on the study of Auditory Evoked Potentials (AEPs) recorded on the scalp of healthy humans and epileptic patients. The protocol allowed us to compare AEPs recorded with or without exposure to RFs. To get a reference, a control session was also introduced. In this study, the correlation coefficients computed between AEPs, as well as the correlation coefficients between spectra of AEPs were investigated to detect a possible difference due to RFs. A difference in the correlation coefficients computed in control and experimental sessions was observed, but it was difficult to deduce the effect of RFs on human health. PMID- 15887253 TI - Localized effects of microwave radiation on the intact eye lens in culture conditions. AB - A novel experimental system was used to investigate the localized effects of microwave radiation on bovine eye lenses in culture for over 2 weeks. Using this setup, we found clear evidence that this radiation has a significant impact on the eye lens. At the macroscopic level, it is demonstrated that exposure to a few mW at 1 GHz for over 36 h affects the optical function of the lens. Most importantly, self-recovery occurs if the exposure is interrupted. At the microscopic level, close examination of the lens indicates that the interaction mechanism is completely different from the mechanism-causing cataract via temperature increase. Contrary to the latter's effect, that is particularly pronounced in the vicinity of the sutures and it is assumed to be a result of local friction between the edges of the fibers consisting the lens. Even if macroscopically the lens has recovered from the irradiation, microscopically the indicators of radiation impact remain. PMID- 15887254 TI - Influence of pulsed electromagnetic field with different pulse duty cycles on neurite outgrowth in PC12 rat pheochromocytoma cells. AB - The influence of low frequency (50 Hz repetition rate) pulsed electromagnetic field (EMF) on PC12 cell neurite outgrowth in vitro was investigated in this study. We studied the percentage of neurite bearing cells, average length of neurites, and directivity of neurite outgrowth in PC12 cells cultured for 96 h in the presence of nerve growth factor (NGF). PC12 cells were exposed in one incubator to pulsed EMF at 1.36 mT (peak value) generated by a pair of Helmholtz coils, and the control samples were placed in another identical incubator. We found that the pulse duty cycle had significant effect on neurite outgrowth. Low (10%) pulse on-time significantly inhibited the percentage of neurite bearing cells, but at the same time increased the average length of neurites, while 100% on-time (DC) had exactly the opposite effects. Furthermore, we found that neurites were prone to extend along the direction of pulsed EMF with 10% pulse on time. Our studies show that neurite outgrowth in PC12 cells is sensitive to the pulse duty and this sensitivity was associated with NGF concentration. PMID- 15887255 TI - Resting EEG effects during exposure to a pulsed ELF magnetic field. AB - Continuing evidence suggests that extremely low frequency magnetic fields (ELF MFs) can affect animal and human behavior. We have previously demonstrated that after a 15 min exposure to a pulsed ELF MF, with most power at frequencies between 0 and 500 Hz, human brain electrical activity is affected as measured by electroencephalography (EEG), specifically within the alpha frequency (8-13 Hz). Here, we report that a pulsed ELF MF affects the human EEG during the exposure period. Twenty subjects (10 males; 10 females) received both a magnetic field and a sham session of 15 min in a counterbalanced design. Analysis of variance (ANOVA) revealed that alpha activity was significantly lower over the occipital electrodes (O1, Oz, O2) [F(1,16) = 5.376, P < .01, eta2 = 0.418] after the first 5 min of magnetic field exposure and was found to be related to the order of exposure (MF-sham vs. sham-MF). This decrease in alpha activity was no longer significant in the 1st min post-exposure, compared to sham (P > .05). This study is among the first to assess EEG frequency changes during a weak (+/-200 microTpk), pulsed ELF MF exposure. PMID- 15887256 TI - Controversial cytogenetic observations in mammalian somatic cells exposed to extremely low frequency electromagnetic radiation: a review and future research recommendations. AB - During the years 1990-2003, a large number of investigations were conducted using animals, cultured rodent and human cells as well as freshly collected human blood lymphocytes to determine the genotoxic potential of exposure to nonionizing radiation emitted from extremely low frequency electromagnetic fields (EMF). Among the 63 peer reviewed scientific reports, the conclusions from 29 studies (46%) did not indicate increased damage to the genetic material, as assessed from DNA strand breaks, incidence of chromosomal aberrations (CA), micronuclei (MN), and sister chromatid exchanges (SCE), in EMF exposed cells as compared with sham exposed and/or unexposed cells, while those from 14 investigations (22%) have suggested an increase in such damage in EMF exposed cells. The observations from 20 other studies (32%) were inconclusive. This study reviews the investigations published in peer reviewed scientific journals during 1990-2003 and attempts to identify probable reason(s) for the conflicting results. Recommendations are made for future research to address some of the controversial observations. PMID- 15887257 TI - Optimization of pulsed electromagnetic field therapy for management of arthritis in rats. AB - Studies were undertaken to find out the effects of low frequency pulsed electromagnetic field (PEMF) in adjuvant induced arthritis (AIA) in rats, a widely used model for screening potential therapies for rheumatoid arthritis (RA). AIA was induced by an intradermal injection of a suspension of heat killed Mycobacterium tuberculosis (500 mug/0.1 ml) into the right hind paw of male Wistar rats. This resulted in swelling, loss of body weight, increase in paw volume as well as the activity of lysosomal enzymes viz., acid phosphatase, cathepsin D, and beta-glucuronidase and significant radiological and histological changes. PEMF therapy for arthritis involved optimization of three significant factors, viz., frequency, intensity, and duration; and the waveform used is sinusoidal. The use of factorial design in lieu of conventional method resulted in the development of an ideal combination of these factors. PEMF was applied using a Fransleau-Braunbeck coil system. A magnetic field of 5 Hz x 4 muT x 90 min was found to be optimal in lowering the paw edema volume and decreasing the activity of lysosomal enzymes. Soft tissue swelling was shown to be reduced as evidenced by radiology. Histological studies confirmed reduction in inflammatory cells infiltration, hyperplasia, and hypertrophy of cells lining synovial membrane. PEMF was also shown to have a membrane stabilizing action by significantly inhibiting the rate of release of beta-glucuronidase from lysosomal rich and sub-cellular fractions. The results indicated that PEMF could be developed as a potential therapy in the treatment of arthritis in humans. PMID- 15887258 TI - Effect of weak, interrupted sinusoidal low frequency magnetic field on neural regeneration in rats: functional evaluation. AB - A study of the effect of weak, interrupted sinusoidal low frequency magnetic field (ISMF) stimulation on regeneration of the rat sciatic nerve was carried out. In the experiment, 60 Wistar rats were used: 24 rats underwent unilateral sciatic nerve transection injury and immediate surgical nerve repair, 24 rats underwent unilateral sciatic nerve crush injury, and the remaining 12 rats underwent a sham surgery. Half of the animals (n = 12) with either sciatic nerve lesion were randomly chosen and exposed between a pair of Helmholtz coils for 3 weeks post-injury, 4 h/day, to an interrupted (active period to pause ratio = 1.4 s/0.8 s) sinusoidal 50 Hz magnetic field of 0.5 mT. The other half of the animals (n = 12) and six rats with sham surgery were used for two separate controls. Functional recovery was followed for 6 weeks for the crush injuries and 7(1/2) months for the transection injuries by video assisted footprint analysis in static conditions and quantified using a recently revised static sciatic index (SSI) formula. We ascertained that the magnetic field influence was weak, but certainly detectable in both injury models. The accuracy of ISMF influence detection, determined by the one-way repeated measures ANOVA test, was better for the crush injury model: F(1, 198) = 9.0144, P = .003, than for the transection injury model: F(1, 198) = 6.4826, P = .012. The Student-Newman-Keuls range test for each response day yielded significant differences (P < .05) between the exposed and control groups early in the beginning of functional recovery and later on from the points adjacent to the beginning of the plateau, or 95% of functional recovery, and the end of observation. These differences probably reflect the ISMF systemic effect on the neuron cell bodies and increased and more efficient reinnervation of the periphery. PMID- 15887259 TI - Complex chromosome rearrangement and recombinant balanced translocation in a mother and a daughter with the same phenotypic abnormalities. AB - We report on the diagnosis of a complex chromosome rearrangement in a mother and the transmission of a simplified translocation in her fetus. The mother had mental retardation, short stature, facial dysmorphism, and hydronephrosis, but was never investigated before she was pregnant. A blood sample was taken for karyotyping at the time of amniocentesis for advanced maternal age. The mother's karyotype revealed two translocations involving chromosome 5, chromosome 16 twice, and chromosome 20 as follow: 46,XX,t(5;16;20)(5pter-->5q11.2::16q12.1- >16q23::20p11.2-->20pter;16pter-->16q12.1::5q11.2-->5qter;16qter-->16q23::20p11.2 ->20qter). The amniocentesis revealed a female karyotype with an apparently balanced translocation: 46,XX,t(16;20)(q23;p11.2). The translocation of the fetus probably resulted from a meiotic recombination between the derived 5 and the normal 16 in the mother. The baby was born and presented the same facial dysmorphism and hydronephrosis. The simplification of a complex rearrangement through recombination into a balanced product has only been rarely described and it is to our knowledge the first time that both the carrier of the complex rearrangement and her descendant with a simplified rearrangement share phenotypic abnormalities. PMID- 15887260 TI - Thresholds for 60 Hz magnetic field stimulation of peripheral nerves in human subjects. AB - The goal of the research reported here is to narrow the range of uncertainty about peripheral nerve stimulation (PNS) thresholds associated with whole body magnetic field exposures at 50/60 Hz. This involved combining PNS thresholds measured in human subjects exposed to pulsed magnetic gradient fields with calculations of electric fields induced in detailed anatomical models of the body by that same exposure system. PNS thresholds at power frequencies (50/60 Hz) can be predicted from these data due to the wide range of pulse durations (70 mus to 1 ms), the length of the pulse trains (several tens of ms), and the exposure of a large part of the body to the magnetic field. These data together with the calculations of the rheobase electric field exceeded in 1% (E(1%)) of two anatomical body models, lead to a median PNS detection threshold of 47.9 +/- 4.4 mT for a uniform 60 Hz magnetic field exposure coronal to the body. The threshold for the most sensitive 1% of the population is about 27.8 mT. These values are lower than PNS thresholds produced by magnetic fields with sagittal and vertical orientations or nonuniform exposures. PMID- 15887261 TI - Mayer-Rokitansky-Kuster-Hauser anomaly and its associated malformations. AB - Mayer-Rokitansky-Kuster-Hauser (MRKHA) is a malformation complex comprising absent vagina and absent or rudimentary uterus. The aim of our study was to describe the type and frequency of anomalies associated with the MRKHA. Between 1975 and 2002, 25 patients with a diagnosis of MRKHA were identified at the Mayo Clinic. These charts were reviewed retrospectively. Two of the 25 patients were found to have absence of one ovary and two patients had unilateral oophorectomies performed for benign cysts. Four patients had minor anomalies and two had digital anomalies. The frequency of scoliosis (20%), unilateral renal agenesis (28%), non vertebral skeletal anomalies (16%) and of the MURCS association (Mullerian agenesis, renal agenesis/ectopia and cervical somite dysgenesis), 16%, was similar compared to that of other published studies. Vertebral abnormalities were found more frequently in our patients (44%). Four patients had cardiac defects, an anomaly not previously described, including truncus arteriosus, patent ductus arteriosus and patent foramen ovale, mitral valve prolapse, and mild mitral regurgitation. We document cardiac anomalies in 16% of our patients with MRKHA suggesting that a search for associated anomalies including cardiac defects is indicated in all such patients. PMID- 15887263 TI - Eponymous Jacobsen syndrome: mapping the breakpoints of the original family suggests an association between the distal 1.1 Mb of chromosome 21 and osteoporosis in Down syndrome. PMID- 15887264 TI - Xq chromosome duplication in males: clinical, cytogenetic and array CGH characterization of a new case and review. AB - Males with duplications within the long arm of the X chromosome are rare and most cases are inherited from a maternal heterozygote. We report a male with a de novo Xq duplication and review of the literature. The proband was ascertained prenatally after an abnormal expanded alpha-fetoprotein (AFP) screen and abnormal ultrasound findings. Chromosome analysis on amniocyte and subsequent peripheral blood lymphocyte cultures showed a male karyotype containing additional material on the long arm of the X chromosome. Fluorescence in situ hybridization with an X chromosome whole chromosome paint probe showed that the additional material was derived from the X chromosome, interpreted as a dup(X)(q13.3q24). Further characterization of the duplication by array CGH showed a duplication size between 30-44 Mb as determined by the map position of the flanking clones on the array, and refined the breakpoints of the duplicated region to Xq21.32 --> Xq25. At birth, the proband had multiple craniofacial abnormalities, musculoskeletal anomalies, bilateral cryptorchidism with scrotal hypoplasia, conductive hearing loss, and profound generalized hypotonia despite normal birthweight, length, and head circumference. Although data regarding Xq duplications in males are limited, a clear pattern of characteristic features can be discerned as illustrated in the present case and confirmed in our literature review. Mental, psychomotor and growth retardation, as well as, craniofacial anomalies, muscle hypotonia, hypoplastic genitalia, cryptorchidism, feeding difficulties, and endocrine dysfunction are all significant issues in these individuals. PMID- 15887265 TI - Conformation of gramicidin a in water: inference from analysis of hydrogen/deuterium exchange behavior by matrix assisted laser desorption ionization mass spectrometry. AB - Gramicidin A (the major component of gramicidin D) is a highly hydrophobic peptide with very little solubility in water. Hence, the conformation of this peptide has been extensively investigated in organic solvents and model membranes, but not in water. The peptide adopts a beta6.3-helical conformation in the monomeric and dimeric forms. We have investigated the conformation of gramicidin A in water by monitoring hydrogen-deuterium exchange by matrix assisted laser desorption ionization-time-of-flight mass spectrometry. Our results indicate that gramicidin A is monomeric and exists in a highly folded conformation. The metal ion bound forms are clearly discernible in the monomers. The presence of the dimeric form is not observed. It is unlikely this is due to the operating conditions or the method used, as both hetero- and homodimers in gramicidin D are detected when methanol is used as a solvent. The present study also establishes that the linear gramicidins retain a history of solvent environment when ions are generated by matrix-assisted laser desorption ionization and analyzed by time-of-flight. PMID- 15887266 TI - Cockroach homologs of praying mantis peripheral auditory system components. AB - This study identifies the cuticular metathoracic structures in earless cockroaches that are the homologs to the peripheral auditory components in their sister taxon, praying mantids, and defines the nature of the cuticular transition from earless to eared in the Dictyoptera. The single, midline ear of mantids comprises an auditory chamber with complex walls that contain the tympana and chordotonal transduction elements. The corresponding area in cockroaches, between the furcasternum and coxae, has many socketed hairs arranged in discrete fields and the Nerve 7 chordotonal organ, the homolog of the mantis tympanal organ. The Nerve 7 chordotonal organ attaches at the apex of the lateral ventropleurite (LVp), which has the same shape and general structure as an auditory chamber wall. High-speed video shows that when the coxa moves toward the midline, the LVp rotates medially to stimulate socketed hairs, and also moves like a triangular hinge giving the chordotonal organ maximal in-out stimulation. Formation of the mantis auditory chamber from the LVp and adjacent structures would involve only enlargement, a shift toward the midline, and a mild rotation. Almost all proprioceptive function would be lost, which may constitute the major cost of building and maintaining the mantis ear. Isolation from leg movement dictates the position of the mantis ear in the midline and the rigid frame, formed by the cuticular knobs, which protects the chordotonal organs. PMID- 15887267 TI - Progressive loss of the spongiotrophoblast layer of Birc6/Bruce mutants results in embryonic lethality. AB - We have generated a mouse line with a mutant allele of the mouse Bruce/Birc6 gene induced by gene trap mutagenesis. Based on its structural features, Bruce is a member of the family of apoptosis inhibitor proteins (IAPs). This mutation leads to a truncated transcript and protein and results in a complete loss of the wildtype Bruce protein. Bruce mutant mice die from a progressive loss of their placental spongiotrophoblast layer between day 11.5 and 14.5 of embryonic development. The cause of the Bruce homozygous mutant phenotype is a lack of proliferation of spongiotrophoblast cells in the developing placenta. In contrast to in vitro data, which indicate a function for Bruce in apoptosis inhibition, the in vivo results presented here suggest instead a role for Bruce in cell division. PMID- 15887268 TI - Unzipping DNA from the condensed globule state-effects of unraveling. AB - We have studied theoretically the unzipping of a double-stranded DNA from a condensed globule state by an external force. At constant force, we found that the double-stranded DNA unzips an at critical force Fc and the number of unzipped monomers M goes as M approximately (Fc - F)-3, for both the homogeneous and heterogeneous double-stranded DNA sequence. This is different from the case of unzipping from an extended coil state in which the number of unzipped monomers M goes as M approximately (Fc - F)-chi, where the exponent chi is either 1 or 2 depending on whether the double-stranded DNA sequence is homogeneous or heterogeneous, respectively. In the case of unzipping at constant extension, we found that for a double-stranded DNA with a very large number N of base pairs, the force remains almost constant as a function of the extension, before the unraveling transition, at which the force drops abruptly to zero. Right at the unraveling transition, the number of base pairs remaining in the condensed globule state is still very large and goes as N(3/4), in agreement with theoretical predictions of the unraveling transition of polymers stretched by an external force. PMID- 15887269 TI - Del(18)(q12.2q21.1) caused by a paternal sister chromatid rearrangement in a developmentally delayed girl. AB - Monosomy of 18q12.3 has been reported in only 16 cases, in one as a mosaic with a normal cell line. Abnormal behaviour, developmental delay, normal measurements, and minor facial anomalies including ptosis, bilateral epicanthus, strabismus, short and slightly down-slanting palpebral fissures, and full cheeks are characteristic manifestations. We report on a 26-month-old girl with del(18)(q12.2q21.1) and typical phenotype. Microsatellite mediated haplotype analysis showed approximately 12 Mb deletion and demonstrated that the deletion was most likely formed during paternal meiosis by a rearrangement between the grandpaternal sister chromatids. PMID- 15887271 TI - Tumor risk in Beckwith-Wiedemann syndrome: A review and meta-analysis. AB - Beckwith-Wiedemann syndrome (BWS) is an overgrowth syndrome associated with macroglossia, abdominal wall defects, ear anomalies, and an increased risk for embryonic tumors. Reported tumor risk estimates vary between 4% and 21%. It has been hypothesized that tumor predisposition in BWS is related to the imprinting status of the H19 and LIT1 genes on chromosome 11p15. A loss of imprinting (LOI) of H19 implies a higher tumor risk. However, a systematic analysis of available data is lacking. Therefore, we performed a review and meta-analysis of reported associations between the imprinting status of the LIT1 and H19 genes and the risk for tumor development in BWS. Five publications suitable for meta-analysis were identified by electronic database searches. Sufficient data were available for 402 out of 520 patients. Patients were divided into four groups based on the imprinting status of H19 and LIT1: group I with LOI of LIT1 (45%); group II with LOI of H19 (9%); group III with LOI of LIT1 and LOI of H19 (21%); and group IV with normal imprinting patterns (26%). Differences in tumor risk between groups were studied with random effects meta-analysis. Tumors occurred in 55 patients. The odds of tumor development was significantly lower in group I when compared to group II (OR=0.06; 95% CI: 0.02-0.21) and group III (OR=0.12; 95% CI: 0.04-0.37). Tumor risk did not differ significantly between groups II and III (OR=1.40; 95% CI: 0.56-3.50). Compared to group IV, tumor risk was significantly lower in group I (OR=0.33; 95% CI: 0.12-0.87) and higher in groups II (OR=4.0; 95% CI: 1.5-10.4) and III (OR=2.6; 95% CI: 1.2-5.7). Tumor incidence rate for group IV was 10.6% (95% CI: 3.6-17.7). Calculated absolute risks were 3% for group I, 43% for group II, and 28% for group III, respectively. No Wilms tumor was seen in group I. In total, other tumors were seen with comparable frequencies in groups I-III. The results show a strong association between a LOI of H19 and especially Wilms tumor development in BWS. PMID- 15887272 TI - Tel Hashomer camptodactyly syndrome: 12-year follow-up of a Hungarian patient and review. AB - Tel Hashomer camptodactyly syndrome (THCS) was diagnosed in a 4-month-old boy whom we have followed for 12 years. In addition to the characteristic clinical findings, he had preductal coarctation of the aorta, persistent ductus arteriosus, and multiple ventricular septal defects. The electron-microscopic evaluation of his muscle biopsy showed anomalies of the sarcoplasmic reticulum and mitochondria; the organization of the myofibrils was normal. The morphological findings suggested primary or secondary involvement of neuromuscular signal transduction and involvement of mitochondria in the development of the myopathy in this child. PMID- 15887273 TI - Autosomal dominant inheritance of spondyloenchondrodysplasia. AB - Spondyloenchondrodysplasia comprises generalized enchondromatosis with platyspondyly and is thought to be inherited as an autosomal recessive condition. A mother and son are reported with typical features of spondyloenchondrodysplasia. Their similar radiographic and MRI findings are presented. The radiologic appearance of the spine changed over time, illustrating the evolving phenotype of this condition. Transmission from mother to son suggests that dominant pattern of inheritance is possible. A classification of the enchondromatoses is discussed. PMID- 15887274 TI - Cerebral, cerebellar, and colobomatous anomalies in three related males: Sex linked inheritance in a newly recognized syndrome with features overlapping with Joubert syndrome. AB - We present a so far unrecognized X-linked mental retardation syndrome with features overlapping with Joubert syndrome (JBS). Two brothers showed hypotonia, mental retardation, ocular abnormalities with impaired vision and colobomas and a breathing pattern compatible with JBS. Neuroimaging revealed cerebellar vermis hypoplasia and ventriculomegaly. A tentative diagnosis of JBS was made, and autosomal recessive inheritance considered most likely. In a subsequent pregnancy that occurred after artificial donor insemination, ultrasound in the 22nd week revealed a Dandy-Walker malformation and hydrocephaly. At autopsy at 34 weeks of gestation, the male infant showed cerebellar vermis aplasia and abnormalities of the brainstem and cerebral cortex. He was considered to have the same disorder as his two half-brothers. This renders the pedigree highly suggestive of X-linked inheritance. The clinical symptoms of this syndrome resemble JBS. However, the absence of the molar tooth sign and the X-linked inheritance do not support JBS. We propose the name X-linked cerebral-cerebellar-coloboma syndrome to distinguish the two disorders. Differentiation of the two disorders is especially important in genetic counseling, where artificial donor insemination may be considered as a means of reducing the recurrence risk, or when female relatives of the patient are concerned. PMID- 15887275 TI - Are the betaine-homocysteine methyltransferase (BHMT and BHMT2) genes risk factors for spina bifida and orofacial clefts? AB - Abnormalities in folate and/or homocysteine metabolism may adversely influence embryonic development, leading to the birth of infants with a variety of congenital malformations, including neural tube defects (NTDs) and craniofacial abnormalities. Based upon suggestive evidence that periconceptional folic acid supplementation is effective in preventing a significant proportion of the aforementioned birth defects, genetic variation in the folate biosynthetic pathways may influence the infant's susceptibility to these birth defects. The goal of our study was to investigate sequence variations in the betaine homocysteine methyltransferase (BHMT) and betaine-homocysteine methyltransferase (BHMT2) genes as modifiers of risk of spina bifida, cleft palate, and cleft lip and palate. The results of this study indicated that individuals homozygous for the single nucleotide polymorphism R239Q in BHMT did not have elevated risks for spina bifida. Genotype frequencies for the BHMT2 rs626105 polymorphism also did not reveal any elevated risks for spina bifida, and only a modest, imprecise elevation of risk for orofacial clefts. The results of these experiments suggest that variants of the BHMT/BHMT2 genes in infants do not substantially contribute to the risk of spina bifida or orofacial clefts in our study population. PMID- 15887276 TI - No evidence of paternal transmission of fragile X syndrome. PMID- 15887277 TI - Novel mutation in DGUOK in hepatocerebral mitochondrial DNA depletion syndrome associated with cystathioninuria. AB - Mitochondrial depletion syndrome (MDS) refers to a heterogeneous group of mitochondrial disorders characterized by a reduction of the mtDNA copy number in affected tissues. Mutations in DGUOK encoding deoxyguanosine kinase (MIM 601465) cause the hepatocerebral form of MDS (MIM 251880). Cystathioninuria (MIM 219500) can result from mutations in CTH encoding cystathionine gamma lyase (MIM 607657) or can be a secondary finding in several diverse clinical conditions. We present three patients from two apparently unrelated old colony Mennonite families, each of whom had the hepatocerebral form of MDS together with cystathioninuria. Each affected child was homozygous for the novel DGUOK p.D255Y mutation, but had no CTH mutation, indicating that the hepatocerebral form of MDS might be associated with secondary cystathioninuria. PMID- 15887278 TI - Autosomal recessive omodysplasia: early prenatal diagnosis and a possible clue to the gene location. AB - Autosomal recessive omodysplasia (ARO), a rare congenital skeletal dysplasia, is characterized by micromelia and craniofacial anomalies. Upper and lower limbs are affected in contrast to the dominant form in which the lower limbs are normal. Radiographic features include shortening and distal tapering of the humerus and femur, proximal radioulnar diastasis, and anterolateral radial head dislocation. We present a recurrence of ARO in a family, detected on prenatal ultrasound at 13 weeks of gestation. Chromosome analysis of the products of conception and the affected sibling showed a paternally-inherited paracentric inversion of 15q13 to q21.3. Due to similarities in the clinical phenotype between diastrophic dysplasia and this condition, testing for DTDST mutation was performed with no mutation detected. PMID- 15887280 TI - The external ear: more attention to detail may aid syndrome diagnosis and contribute answers to embryological questions. AB - The human pinna has a complex shape and yet the basic components of normal structure are remarkably constant between individuals. It is precocious in its appearance during embryogenesis and it has been subject to many developmental and surgical studies, yet questions remain as to its primary embryogenesis and the causes of its malformations. Unfortunately, the vast majority of clinical reports of syndromes and of individuals with dysmorphic signs provide limited and inadequate description of the external ear. Given the intricate pattern of the pinna, and hence its potential for morphological variation, we think that more attention to the specific description of ear anomalies may lead to a better appreciation of the etiology and embryology of the malformations. Furthermore, in some cases correlation with specific syndromes may provide an aid to diagnosis. Towards those ends this paper reviews some of the controversy concerning the embryology of the pinna, and discusses a number of the better-defined anomalies of the external ear. Although it has been suggested that anomalies of the insertion and orientation of intrinsic muscles of the pinna may be responsible for variations in external ear morphology, we think it likely that in many cases the anomalous insertions may be secondary. PMID- 15887281 TI - Maternal uniparental disomy 14 in a 15-year-old boy with normal karyotype and no evidence of precocious puberty. PMID- 15887282 TI - Immune abnormalities are a frequent manifestation of Kabuki syndrome. AB - Kabuki syndrome (KS) is associated with multiple organ system involvement. Characteristic features include long palpebral fissures with everted lower lids, prominent ears, skeletal abnormalities, mental retardation, and short stature. An increased incidence of infection has been reported in KS, and a few patients have been noted to have immune defects. However, the frequency and severity of the immune deficiency has not been clearly defined. Immunologic evaluation of 19 consecutive individuals with KS was performed at The Children's Hospital of Philadelphia. Decreased IgA levels were noted in 15/19 individuals (79%), 2 of whom had undetectable levels. Eight patients (42%) also had low total IgG levels. Specific IgG subclass abnormalities were found in 6 of 13 patients evaluated. IgM levels were less frequently decreased. One patient failed to generate anti tetanus antibodies despite immunization. This study suggests that hypogammaglobulinemia is a frequent finding in children with KS. The pattern of antibody abnormalities seen in children with KS resembles common variable immune deficiency (CVID). Due to this increased susceptibility to infection, children with KS should have immunologic evaluations at the time of diagnosis in order to reduce preventable morbidity and mortality. PMID- 15887283 TI - A comparative study of endothelial cell markers expressed in chronically inflamed human tissues: MECA-79, Duffy antigen receptor for chemokines, von Willebrand factor, CD31, CD34, CD105 and CD146. AB - Endothelial cells play a central role in chronic inflammation: for example, they express adhesion molecules and present chemokines leading to enhanced leukocyte recruitment into tissues. Numerous markers of endothelial cells have been reported but there has been a lack of comparative data on their specificity. The present study compared the specificity of seven endothelial cell markers in the rheumatoid synovium and the colon of patients with Crohn's disease. These markers were: the sulphated epitope MECA-79, the Duffy antigen receptor for chemokines (DARC), von Willebrand factor, CD31 (PECAM-1), CD34, CD105 (endoglin) and CD146. MECA-79, DARC and von Willebrand factor showed a specific endothelial cell distribution. MECA-79, which recognizes sulphated ligands for leukocyte adhesion receptor L-selectin (CD62L), was selective for a subset of venules in highly inflamed tissue and was present in rheumatoid but not control osteoarthritic synovia. DARC was also specific for venules but had a more widespread distribution than MECA-79, and was present in rheumatoid and control synovia. The other markers all labelled endothelial cells in venules, arterioles and capillaries. However, they also localized to other cell types. For example, CD34 stained fibroblasts, CD146 was expressed by the pericytes and smooth muscle cells of vessel walls and CD31 and CD105 labelled a broad range of cell types. PMID- 15887284 TI - Genetic regulation of osteoarthritis: A QTL regulating cranial and caudal acetabular osteophyte formation in the hip joint of the dog (Canis familiaris). PMID- 15887285 TI - Extending logistic regression to model diffuse interactions. AB - In an observational study focussed on association between a health outcome and numerous explanatory variables, the question of interactions can be problematic. Commonly, logistic regression of the outcome on the explanatory variables might be employed. Such modelling often includes an attempt to select some pairwise product interaction terms, from amongst the many such possible pairs. For several reasons, however, this can be unsatisfying. Here we consider a different approach based on a parsimonious extension of a logistic regression model without interaction terms. This extension permits an overall synergism or antagonism in how the explanatory variables combine to associate with the outcome, without any attempt to identify specific variables which give rise to interactive behaviour. We call this diffuse interaction. We elucidate some simple properties of the diffuse interaction model, and give an example of its application to epidemiological data. We also consider asymptotic behaviour in a restricted case of the model, to gain some insight into how well this kind of interaction can be detected from data. PMID- 15887286 TI - Distinguishing Pacman dysplasia from mucolipidosis II: comment on Saul et al. [2005]. PMID- 15887287 TI - The transactivating isoforms of p63 are overexpressed in high-grade follicular lymphomas independent of the occurrence of p63 gene amplification. AB - p63 is a p53-related gene mapping to 3q28 that codes for multiple mRNA transcripts with (TA-p63) or without (DeltaN-p63) transactivating effects on genes that promote cell differentiation and apoptosis. We analysed p63 alterations by immunohistochemistry, quantitative real-time RT-PCR and FISH in a series of 45 follicular lymphomas (FL). None of the tumours showed immunoreactivity for the p40 antibody, which recognizes only the truncated isoforms of p63, or DeltaN-p63 mRNA expression. Immunoreactivity for the 4A4 antibody, which recognizes both the transactivating and the truncated p63 isoforms, was found in 5 +/- 5.5%, 6.85 +/- 4.88% and 33.2 +/- 22.31% of grade I, II and III FL cells, respectively (p < 0.0001). Quantitative RT-PCR analysis showed that all cases but one had TA-p63 mRNA levels higher than non-neoplastic lymphocytes, and that TA-p63 mRNA expression correlated significantly (r = 0.9194, p < 0.0001) with the prevalence of p63 immunoreactivity. FISH extra signals for the p63 gene were found in seven (23.3%) of the 30 cases analysed (0/6 grade I, 2/15 grade II and 5/9 grade III; p = 0.01937). Further hybridizations showed a pattern highly suggestive of chromosome 3 polysomy in six cases. One of these cases also bore extra copies of the p63 and bcl-6 genes. Co localization of p63 and IgH signals was found in one case. No association between the prevalence of p63 immunoreactivity and extra p63 gene signals was detectable when the cases were dichotomized according to a p63 immunoreactivity threshold of 10%. Our data suggest that TA-p63 is overexpressed in high-grade FL, possibly independent of the occurrence of gene abnormalities, and that it may be involved in the highly complex mechanism of regulation of apoptosis of FL cells. PMID- 15887288 TI - RASSF1A promoter methylation and 3p21.3 loss of heterozygosity are features of foregut, but not midgut and hindgut, malignant endocrine tumours. AB - The Ras-association domain family 1A (RASSF1A) tumour suppressor gene is inactivated in a variety of solid tumours, usually by epigenetic silencing of the promoter and/or allelic loss of its locus at 3p21.3. RASSF1A induces cell cycle arrest through inhibition of cyclin D1 accumulation. In this work, 62 endocrine tumours from different sites in the gut were investigated for methylation of the RASSF1A promoter using the polymerase chain reaction, the presence of 3p21.3 deletions by loss of heterozygosity analysis, and cyclin D1 expression by immunohistochemistry. Methylation was found in 20/62 (32%) cases and was restricted to foregut tumours; deletion at 3p21.3 was found in 15/58 (26%) informative cases and restricted to malignant foregut tumours; cyclin D1 hyper expression was found in 31/58 (53%) cases and correlated with RASSF1A methylation. Our data suggest that RASSF1A is involved in the development of endocrine tumours derived from the foregut only, and that the presence of both RASSF1A methylation and 3p21.3 deletion is associated with malignancy. These results may provide a rationale for foregut-targeted therapy for aggressive endocrine carcinomas entailing the use of demethylating agents. PMID- 15887289 TI - Prenatal mucolipidosis type II (I-cell disease) can present as Pacman dysplasia. AB - Pacman dysplasia has been previously reported to be a lethal skeletal dysplasia with epiphyseal stippling and osteoclastic overactivity. We report on a sibling of a fetus previously reported as Pacman dysplasia. This infant has a clinical course consistent with mucolipidosis type II (I-cell disease) along with confirmatory biochemical, cytologic, and radiographic evidence. This case expands the phenotypic spectrum of mucolipidosis type II. Having redefined the diagnosis in one of the original cases of Pacman dysplasia, we suggest that what is called Pacman dysplasia could very well be Mucolipidosis type II (ML-II) in other published reports. PMID- 15887291 TI - High numbers of tumour-infiltrating activated cytotoxic T lymphocytes, and frequent loss of HLA class I and II expression, are features of aggressive B cell lymphomas of the brain and testis. AB - Loss of both HLA class I and class II expression in B cell lymphomas is a mechanism of escape from a cytotoxic T lymphocyte (CTL) immune response and will therefore give a strong selective survival advantage in tumours expressing strong immunogenic antigens. We investigated loss of HLA expression using specific antibodies on tissue sections from 254 B cell lymphomas originating from nodal and different extranodal sites in relation to numbers of tumour-infiltrating T cells. Complete loss of HLA class I and II was observed in a minority of the nodal, stomach, and skin lymphomas but in the majority of the lymphomas originating from the testis and the CNS. Interestingly, relatively high percentages of activated CTLs were detected in both primary testicular and CNS lymphomas compared to lymphomas at other sites, with highest percentages in the testis (p < 0.0001). We conclude that loss of both HLA class I and II expression occurs very frequently in lymphomas originating from the testis and the CNS as compared to nodal and some other extranodal sites. The presence of high percentages of activated CTLs in the testicular and CNS lymphomas suggests that loss of HLA expression provides a strong growth advantage for lymphoma cells in these immune-privileged sites. PMID- 15887292 TI - IgV H mutations in blastoid mantle cell lymphoma characterize a subgroup with a tendency to more favourable clinical outcome. AB - Mantle cell lymphoma (MCL) is associated with a very unfavourable clinical course. This is particularly true for mantle cell lymphoma of the blastoid subtype (MCL-b). In order to define prognostic factors, we analysed the impact of immunoglobulin heavy chain variable (IgV H) gene somatic hypermutations on clinical outcome in a series of 21 cases of morphologically, phenotypically, and genotypically well-characterized MCL-b. Testing and estimation were performed using log-rank statistics and displayed on Kaplan-Meier graphs. Thirteen of 21 cases of MCL-b revealed a homology rate of > or = 99% compared to IgV H germ-line sequences in the databases and were scored as non-mutated. Eight of 21 cases (38%) of MCL-b were mutated. In MCL-b the mutation frequency was usually low and the mutation pattern was only rarely antigen-selected, in contrast to a control group of 11 cases with morphologically almost identical, but phenotypically and genotypically clearly distinguishable, diffuse large B cell lymphoma, derived, most likely, from germinal centre B cells. In our series of 21 MCL-b, positive IgV H mutational status, irrespective of varying homology thresholds, had no statistically significant prognostic impact on event-free or overall survival. However, mutated MCL-b tended to present more frequently at an earlier stage and without bone marrow involvement and to show lower rates of relapse and death, resulting in a more favourable clinical outcome. PMID- 15887293 TI - Polymorphisms in DNA repair genes as risk factors for spina bifida and orofacial clefts. AB - Repairing DNA damage is critical during embryogenesis because development involves sensitive periods of cell proliferation, and abnormal cell growth or death can result in malformations. Knockout mouse experiments have demonstrated that disruption of DNA repair genes results in embryolethality and structural defects. Studies using mid-organogenesis rat embryos showed that DNA repair genes were variably expressed. It is hypothesized that polymorphisms that alter the functionality of DNA repair enzymes may modify the risk of malformations. We conducted a case-control analysis to investigate the relationship between DNA repair gene polymorphisms and the risk of spina bifida and oral clefts. Newborn screening blood spot DNA was obtained for 250 cases (125 spina bifida, 125 oral clefts) identified by the California Birth Defects Monitoring Program, and 350 non-malformation controls identified from birth records. Six single nucleotide polymorphisms of five DNA repair genes representing three distinct repair pathways were interrogated including: XRCC1 (Arg399Gln), APE1 (Asp148Glu), XRCC3 (Thr241Met), hOGG1(Ser326Cys), XPD (Asp312Asn, Lys751Gln). Elevated or decreased odds ratios (OR, adjusted for race/ethnicity) for spina bifida were found for genotypes containing at least one copy of the variant allele for XPD [751Gln, OR = 1.62; 95% confidence interval (CI) = 1.05-2.50] and APE 148 (OR = 0.58; CI = 0.37-0.90). A decreased risk of oral clefts was found for XRCC3 (OR = 0.62; CI = 0.39-0.99) and hOGG1 (326 Cys/Cys, OR = 0.22; CI = 0.06-0.78). This study suggested that polymorphisms of DNA repair genes, representing different major repair pathways, may affect risk of two major birth defects. Future, larger studies, examining additional repair genes, birth defects, and interaction with exposures are recommended. PMID- 15887294 TI - c-Kit immunophenotyping and metalloproteinase expression profiles of mast cells in interstitial lung diseases. AB - Diverse interstitial lung diseases (ILD) demonstrate mesenchymal infiltration by an abundance of activated mast cells whose role in parenchymal fibrogenesis remains unclear. Since mast cells differentiate in a dynamic, tissue-specific manner via signals transduced by c-Kit receptor, we examined the effect of ILD microenvironments on c-Kit expression and metalloproteinase phenotypes of mesenchymal mast cell populations. Immunohistochemical and flow cytometric analyses characterized surface expression of c-Kit on mast cells in tissues obtained from patients with idiopathic pulmonary fibrosis, systemic sclerosis, sarcoidosis, and lymphangioleiomyomatosis, thus identifying a unique immunophenotype not shared by normal lung mast cells. Isolation of c Kit+/FcepsilonRI+/CD34- mast cells via immunocytometric sorting of heterogeneous cell populations from mechanically disaggregated lung tissues permitted analysis of gene expression patterns by two-step real-time polymerase chain reaction. Transcriptional profiling identified expression of c-Kit and the neutral serine proteases, tryptase and chymase, establishing the identity of sorted populations as mature mast cells. Mast cells harvested from ILD tissues demonstrated characteristic metalloproteinase phenotypes which included expression of matrix metalloproteinase (MMP)-1 and a disintegrin and metalloproteinase (ADAM)-9, -10, and -17. Immunohistochemical co-localization guided by gene profiling data confirmed expression of chymase, MMP-1, and ADAM-17 protein in subpopulations of mast cells in remodelling interstitium. Gene profiling of harvested mast cells also showed increased transcript copy numbers for TNFalpha and CC chemokine receptor 2, which play critical roles in lung injury. We conclude that ILD microenvironments induce unique c-Kit receptor and metalloproteinase mast cell phenotypes. PMID- 15887295 TI - SAC-ing mitotic errors: how the spindle assembly checkpoint (SAC) plays defense against chromosome mis-segregation. PMID- 15887296 TI - Meta-analysis of genetic studies using Mendelian randomization--a multivariate approach. AB - In traditional epidemiological studies the association between phenotype (risk factor) and disease is often biased by confounding and reverse causation. As a person's genotype is assigned by a seemingly random process, genes are potentially useful instrumental variables for adjusting for such bias. This type of adjustment combines information on the genotype-disease association and the genotype-phenotype association to estimate the phenotype-disease association and has become known as Mendelian randomization. The information on genotype-disease and genotype-phenotype may well come from a meta-analysis. In such a synthesis, a multivariate approach needs to be used whenever some studies provide evidence on both the genotype-phenotype and genotype-disease associations. This paper presents two multivariate meta-analytical models, which differ in their treatment of the heterogeneities (between-study variances). Heterogeneities on the genotype phenotype and genotype-disease associations may be highly correlated, but a multivariate model that parameterizes the heterogeneity directly is difficult to fit because that correlation is poorly estimated. We advocate an alternative model that treats the heterogeneities on genotype-phenotype and phenotype-disease as being independent. This model fits readily and implicitly defines the correlation between the heterogeneities on genotype-phenotype and genotype disease. We show how either maximum likelihood or a Bayesian approach with vague prior distributions can be used to fit the alternative model. PMID- 15887297 TI - Analysis of force generation during flagellar assembly through optical trapping of free-swimming Chlamydomonas reinhardtii. AB - Many studies have used velocity measurements, waveform analyses, and theoretical flagella models to investigate the establishment, maintenance, and function of flagella of the biflagellate green algae Chlamydomonas reinhardtii. We report the first direct measurement of Chlamydomonas flagellar swimming force. Using an optical trap ("optical tweezers") we detect a 75% decrease in swimming force between wild type (CC124) cells and mutants lacking outer flagellar dynein arms (oda1). This difference is consistent with previous estimates and validates the force measurement approach. To examine mechanisms underlying flagella organization and function, we deflagellated cells and examined force generation during flagellar regeneration. As expected, fully regenerated flagella are functionally equivalent to flagella of untreated wild type cells. However, analysis of swimming force vs. flagella length and the increase in force over regeneration time reveals intriguing patterns where increases in force do not always correspond with increases in length. These investigations of flagellar force, therefore, contribute to the understanding of Chlamydomonas motility, describe phenomena surrounding flagella regeneration, and demonstrate the advantages of the optical trapping technique in studies of cell motility. PMID- 15887299 TI - Omphalocele, advanced maternal age, and fetal morbidity outcomes. AB - In this study we wanted to determine if the risk for adverse neonatal outcome among omphalocele-affected fetuses is increased among older gravidas. This was a retrospective cohort study on live-born infants with omphalocele delivered in New York State from 1983 through 1999. We compared infants of older (>or=35 years) with those of younger (<35 years) mothers with respect to the following fetal morbidity indices: low birth weight and very low birth weight, preterm and very preterm, and small for gestational age. We used adjusted odds ratios to approximate relative risks. Data on a total of 1,010 infants with omphalocele were analyzed. Mean gestational age and birth weight were similar in both maternal age categories: mean+/-standard deviation (SD) for infants with omphalocele born to older mothers=37.4 weeks+/-3.9 versus 38.0 weeks+/-5.1 for those of younger mothers (P=0.2); mean birth weights+/-SD for infants with omphalocele born to older mothers=2,813+/-871.1 versus 2,958+/-809.9 for those of younger mothers (P=0.08). Also, the two maternal age sub-groups did not differ with respect to the fetal morbidity outcome: low birth weight (OR=0.95; 95% CI=0.60-1.51), very low birth weight (OR=0.78; 95% CI=0.36-1.69), preterm (OR=0.95; 95% CI=0.58-1.57), very preterm (OR=0.73; 95% CI=0.34-1.58), and SGA (OR=1.00; 95% CI=0.44-2.27). Thus, advanced maternal age does not appear to be a risk factor for fetal morbidity outcomes among omphalocele-affected fetuses. This information is potentially useful in counseling affected parents. PMID- 15887300 TI - Cockayne syndrome: the developing phenotype. AB - Cockayne syndrome is a rare autosomal recessive condition comprising microcephaly, "cachectic dwarfism" and progressive neurological degeneration. We present a 21-year-old woman who was not diagnosed with Cockayne syndrome type I until she was 21 years old. Family photographs demonstrated that the phenotype of Cockayne syndrome did not become evident until she was 8 years old. She had severe microcephaly, micrognathia, protruding ears, dental overcrowding with caries, progressive spastic quadriparesis, and severe developmental regression. Her head computed tomography (CT) showed bilateral calcification of the globus pallidus and global atrophy. The purpose of this clinical report is to alert clinicians to the fact that the phenotypic features of Cockayne syndrome may be very subtle early in the course of the disease. PMID- 15887301 TI - Sensorineural deafness, abnormal genitalia, synostosis of metacarpals and metatarsals 4 and 5, and mental retardation: description of a second patient and exclusion of HOXD13. AB - In 1988 Pfeiffer and Kapferer reported on a patient with sensorineural deafness, psychomotor delay, hypospadias, cerebral manifestations, and bilateral synostosis of the 4th and 5th metacarpals and metatarsals. Synostosis of the 4th and 5th metacarpals and metatarsals is a very rare defect that has been described as an isolated Mendelian defect, as part of multiple congenital anomaly (MCA) patterns, and in different syndromes. Among a total of 2,023,155 liveborn infants in the Spanish Collaborative Study of Congenital Malformations (ECEMC), we observed only two cases with this type of metacarpal fusion, for a frequency of 1/1,011,577. One had the isolated defect, and the other one that we are describing here, had an MCA pattern similar to that described by Pfeiffer and Kapferer [1988]. We tested HOXD13 but did not find any mutations in exons and intron-exon boundaries. To our knowledge this case is the second one reported with this syndrome. PMID- 15887302 TI - EMX2-independent familial schizencephaly: clinical and genetic analyses. AB - Schizencephaly is a human brain malformation distinguished by full-thickness unilateral or bilateral clefts through the neocortex. Heterozygous mutations in the EMX2 locus are reported to give rise to schizencephaly. However, the comprehensive identification of causative genetic loci is precluded by a lack of large pedigrees and genome-wide linkage analyses. We present here a large Turkish pedigree with three individuals with schizencephaly. The similarity of clinical signs in affected individuals strongly suggests an underlying genetic cause; however, genome-wide linkage analysis rules out EMX2 linkage and instead suggests additional candidate loci. These results indicate that genetic forms of schizencephaly are likely to be heterogeneous. PMID- 15887303 TI - OEIS complex with del(3)(q12.2q13.2). PMID- 15887305 TI - Efficiency comparisons of rank and permutation tests. PMID- 15887309 TI - Spline-based non-parametric regression for periodic functions and its application to directional tuning of neurons. AB - The activity of neurons in the brain often varies systematically with some quantitative feature of a stimulus or action. A well-known example is the tendency of the firing rates of neurons in the primary motor cortex to vary with the direction of a subject's arm or wrist movement. When this movement is constrained to vary in only two dimensions, the direction of movement may be characterized by an angle, and the neuronal firing rate can be written as a function of this angle. The firing rate function has traditionally been fit with a cosine, but recent evidence suggests that departures from cosine tuning occur frequently. We report here a new non-parametric regression method for fitting periodic functions and demonstrate its application to the fitting of neuronal data. The method is an extension of Bayesian adaptive regression splines (BARS) and applies both to normal and non-normal data, including Poisson data, which commonly arise in neuronal applications. We compare the new method to a periodic version of smoothing splines and some parametric alternatives and find the new method to be especially valuable when the smoothness of the periodic function varies unevenly across its domain. PMID- 15887310 TI - On the use of discrete choice models for causal inference. AB - Methodology for causal inference based on propensity scores has been developed and popularized in the last two decades. However, the majority of the methodology has concentrated on binary treatments. Only recently have these methods been extended to settings with multi-valued treatments. We propose a number of discrete choice models for estimating the propensity scores. The models differ in terms of flexibility with respect to potential correlation between treatments, and, in turn, the accuracy of the estimated propensity scores. We present the effects of discrete choice models used on performance of the causal estimators through a Monte Carlo study. We also illustrate the use of discrete choice models to estimate the effect of antipsychotic drug use on the risk of diabetes in a cohort of adults with schizophrenia. PMID- 15887312 TI - Very large container molecules. PMID- 15887313 TI - No increase in peak systolic velocity of the pulmonary veins over time: a potential marker for congenital lobar emphysema. PMID- 15887314 TI - New treatment may eradicate virulent lymphoma. A single course of infections kills cancers, with minimal side-effects; positive responses immediate. PMID- 15887315 TI - Endometrial cancer may need additional treatment. PMID- 15887316 TI - Statins also lengthen life of dialysis patients. PMID- 15887317 TI - Abstracts of the 46th Spring Meeting of the German Society for Experimental and Clinical Pharmacology and Toxicology. March 15-17, 2005, Mainz, Germany. PMID- 15887318 TI - Integrating problem-based learning to interprofessional education curricula using the Corbin and Strauss' illness trajectory model: student evaluations and faculty experiences. PMID- 15887319 TI - Interprofessional care training program pilot project. PMID- 15887320 TI - Interprofessional education for undergraduate health care students throughout their training. PMID- 15887321 TI - Shaping the future for primary care education and training: a collaborative experience. PMID- 15887322 TI - Establishing indicators for biodiversity. PMID- 15887323 TI - Educational programs for deaf students. Schools and programs in the U.S. PMID- 15887325 TI - Educational programs for deaf students. Postsecondary programs. PMID- 15887324 TI - Educational programs for deaf students. Schools and programs in Canada. PMID- 15887326 TI - University and college programs for personnel in deafness. PMID- 15887327 TI - Programs for deaf-blind children and adults. PMID- 15887328 TI - Advocacy, professional, support, and rehabilitation programs. PMID- 15887329 TI - Candida, still number one--what do we know and where are we going from there? AB - Candida species are considered as the most important fungal human pathogens, causing a variety of clinical entities, ranging from superficial, cutaneous mucosal to deep-seated and disseminated infections. A vast body of scientific literature, has been accumulated on these pathogens. A review of the literature and topics for further research are discussed. PMID- 15887330 TI - [Blood pressure variability]. AB - Blood pressure variability is a physiological phenomenon influenced by many internal and external factors. This variability could be also influenced by pathological conditions such as arterial hypertension. Two forms must be mainly distinguished: the blood pressure variability at long but also short-term. The latter could only be studied by continuous recordings. From the initial invasive intraarterial approach, it can nowadays be explored by a non invasive system of beat to beat recordings using the infrared photo plethysmography (the FINAPRES system). In this paper, some important questions will be treated such as the interest of measuring blood pressure variability, its cardiovascular prognosis and how therapeutic tools can be applied when it is increased? PMID- 15887331 TI - An unanswered question. PMID- 15887332 TI - Teratomas as an ANT standard. PMID- 15887333 TI - The burden of proof. PMID- 15887334 TI - Producing non-embryonic organisms for stem cells. PMID- 15887335 TI - The many perils that lie ahead. PMID- 15887336 TI - The substantive issues raised by altered nuclear transfer. PMID- 15887337 TI - Washington insider. The elections and Congress: a continued statement on embryo research? PMID- 15887338 TI - On the use of condoms to protect acquired immunodeficiency syndrome. PMID- 15887339 TI - On "rescuing" frozen embryos: why the decision to do so is moral. PMID- 15887340 TI - The transfer of abandoned frozen embryos--identifying the object of the act. PMID- 15887341 TI - What is chosen in the act of embryo adoption? PMID- 15887342 TI - Practical and moral caveats on heterologous embryo transfer. PMID- 15887343 TI - In defense of transferring heterologous embryos. PMID- 15887344 TI - Can one "rescue" a human embryo? The moral object of the acting woman. PMID- 15887345 TI - Altered nuclear transfer as a morally acceptable means for the procurement of human embryonic stem cells. PMID- 15887346 TI - To the participants in the 19th International Conference of the Pontifical Council for Pastoral Health Care. PMID- 15887347 TI - LSUSD research strengthened with $10.7 million grant from NIH. PMID- 15887348 TI - The new buzzword in dentistry, lasers. PMID- 15887349 TI - Why the tripartite? PMID- 15887351 TI - New practice management software feature makes increasing case acceptance even easier. PMID- 15887350 TI - Dentists play vital role in protecting Louisiana's children. PMID- 15887352 TI - Human or not? A forensic dental case. PMID- 15887353 TI - Special issue: space radiation biology. PMID- 15887354 TI - Proceedings of the 18th annual meeting of Japanese Society for Biological Sciences in Space. PMID- 15887355 TI - Evaluating a Web-based graduate level nursing ethics course: thumbs up or thumbs down? AB - BACKGROUND: This article discusses the course design, experiential findings, and evaluation of a Web-based course for graduate nurses examining professional health care practice and ethical issues. METHOD: Using course input and evaluative data, the article addresses student and teacher perceptions of use of a Web-based delivery mode for graduate level courses and the appropriateness of this ethics course for graduate level students. RESULTS: Evaluative data supported existing knowledge concerning pros and cons of Web-based education. New insights concerning the Web's ability to foster critical thinking skills and supports for continuing ethics education for nurses are reported. All students assessed this ethics course as being of considerable value to their professional practice. CONCLUSION: Students evaluated the pros of Web use for the course as far outweighing the cons. The students' substantial interest in and critical thinking about appropriate knowledge bases to underpin effective ethical decision making strongly supports the need for continuing ethics education for practicing nurses. PMID- 15887356 TI - Lessons learned in developing and delivering Web-based graduate courses: a faculty perspective. AB - The increased demand for graduate distance education by RNs create many challenges for nurse educators. This article describes the trials and tribulations of developing and delivering two Web-based graduate nursing courses at a western Canadian university. The authors describe the background and rationale for developing Web-based courses, the relevant literature, and the lessons learned in planning and delivering these courses. PMID- 15887357 TI - Competence, continuing education, and computers. AB - A survey of RNs in South Dakota was performed to determine their perceived level of competence, the extent to which their continuing nursing education (CNE) needs are being met, and their use of computers for CNE. Nationally certified nurses rated themselves significantly more competent than nurses who are not nationally certified. Fewer than half of the RNs reported their CNE needs were being met despite geographic access to CNE and programs available in their specialty. Three fourths of nurses had computers at home while 76% had computers at work, yet fewer than 20% of nurses used these computers for CNE. PMID- 15887358 TI - Web-based professional education for advanced practice nursing: a consumer guide for program selection. AB - As Web-based education becomes increasingly available, there are more educational opportunities to pursue an advanced practice degree. Despite the increasing number of schools delivering Web-based education courses and programs, evaluation of these programs is primarily in beginning stages. The purposes of this article are first to present a scenario of how a Midwestern nursing school developed, implemented, and continually evaluates Web-based education in its master's program, and second to present a consumer guide for program selection. Elements in the guide are derived from program accrediting agencies, the literature, and distance learning experiences. The consumer's guide is presented for nurses interested in exploring or continuing the completion of an advanced practice degree. PMID- 15887359 TI - A comparison of computer-based and instructor-led training for long-term care staff. AB - Nursing facility administrators must find new and innovative ways to meet the training needs of their staff. The authors faced the challenge of teaching nursing facility staff about fire safety using computer-based training that had been adapted from a print-based/instructor-led (IL) program. This article discusses the effectiveness of the computer-based (CB) training as compared to the effectiveness of the traditional IL format. The CB and IL versions of a fire safety training program were presented to staff of a nursing facility. The 141 participants were randomly assigned to the CB, IL, or control groups. The control group completed a pretest and posttest, but received no training until the study was completed. Both the CB and IL groups significantly increased their knowledge scores from pretest to posttest and both significantly outperformed the control group. How the participants responded to the CB training was also important. Staff reported they enjoyed the CB training and had no difficulty using the computers. Computer-based training can be an effective alternative training technique for this population. PMID- 15887360 TI - Spatial and temporal variation of heavy metals in atmospheric aerosol of Delhi. AB - The levels of Suspended Particulate Matter (SPM) and heavy metals viz. Pb, Cd, Cr, Ni and Fe were measured. Aerosol samples from four different locations in Delhi were collected by High-volume samplers for a period of one year from July 1997 June 1998. Metal concentration was determined by Atomic Absorption Spectrometry. The annual average concentration of SPM in Delhi was found to be 416.34 +/- 223 microg m(-3). The atmospheric aerosol samples were highly enriched with elements viz. Pb and Cd, which originate from various human activities like transportation and industrial processes. Principal Component Analysis (PCA) showed vehicular traffic and industrial emission as the major contributors of metals. The annual average concentration of Pb did not exceed the national standard of 0.75 microg m(-3). PMID- 15887361 TI - Evaluation of potential molecular markers for urban stormwater runoff. AB - Source specificity and persistence of several sulfur-polycyclic aromatic hydrocarbons (S-PAHs), nitro-PAHs (N-PAHs), and triphenylene were examined via analyses of stormwater runoff and wastewater effluent samples and spiked samples upon exposure to sunlight. Samples were collected during the 1997/1998 wet weather season from two major storm channels and four major wastewater treatment plants in southern California. Among the target compounds examined, 2-(4 morpholinyl)benzothiazole, dibenzothiophene, and triphenylene/chrysene were detected in storm runoff only. However, 2-(4-morpholinyl)benzothiazole appeared to degrade rapidly in seawater and sediment after sunlight exposure, which might impede its use as a runoff indicator. Dibenzothiophene and triphenylene also degraded quickly in sunlight-exposed seawater samples, but remained fairly abundant in sediments after six months of exposure to sunlight. They are by far the most promising candidates of urban runoff markers based on the criteria of abundance, source specificity, and persistence, although more research efforts are needed to ensure that no other sources would also contribute significantly to their presence in the aquatic environment. PMID- 15887362 TI - Low levels of sodium and potassium in the water from wetlands in Minnesota that contained malformed frogs affect the rate of Xenopus development. AB - Water samples were collected between 1999 and 2000 from wetlands in Minnesota that contained malformed frogs. The water samples were analyzed for 14 minerals/ions and screened for the presence of biologically active compounds using Xenopus laevis. Results indicated that water from two sites, CWB and ROI2, induced severe retardation with embryo lengths reduced 20% after 96 hr of development. The developmental delay observed with water from ROI2 was alleviated by supplementation with sodium, while both sodium and potassium alleviated the developmental delay observed with water whose mineral content mimicked that of CWB. Seasonal fluctuations in the sodium and potassium content at ROI2 and NEY correlated with changes in the rates of Xenopus development. Xenopus embryos reared on water from ROI2 for 120 hr displayed gut malformations not present in embryos reared on a synthetic media designed to mimic the mineral content of the water from ROI2. Embryos reared on water from ROI2 supplemented with minerals at levels comparable to that routinely employed in the rearing of Xenopus were neither retarded nor malformed. It is proposed that climate driven hydrology may influence the mineral composition at selected wetlands and delay development which may alter window(s) of susceptibility towards biologically active agents and the occurrence of malformed frogs. PMID- 15887363 TI - Monitoring of pesticidal contamination of farmgate vegetables from Hisar. AB - Analysis of 84 farm gate samples of seasonal vegetables carried out on GC-ECD and GC-NPD systems equipped with capillary columns by using a multi residue analytical technique revealed that 26% samples contained residues above MRL values. The contamination was mainly with organophosphates followed by synthetic pyrethroids and organochlorines. Among organophosphates, residues of monocrotophos, quinalphos and chlorpyriphos exceeded the MRL value in 23% samples. Residues of monocrotophos were higher than MRL value in 3 samples of brinjal and one sample each of okra, cauliflower and smooth gourd, of chlorpyriphos in 3 samples of cauliflower and 8 of cabbage and of quinalphos in one sample each of okra and cauliflower. Among synthetic pyrethroids, cypermethrin was the major contaminant and its residue exceeded the MRL value in one sample each of brinjal, okra and cucumber. Only aldicarb among the carbamates was detected in potato. The residues of some organochlorines, i.e. HCH, DDT and endosulfan were found in all the samples but did not exceed the tolerence limit. PMID- 15887364 TI - Gold concentrations in abiotic materials, plants, and animals: a synoptic review. AB - Gold (Au) is ubiquitous in the environment and mined commercially at numerous locations worldwide. It is also an allergen that induces dermatitis in sensitive individuals. Gold concentrations were comparatively elevated in samples collected near gold mining and processing facilities, although no data were found for birds and non-human mammals. Maximum gold concentrations reported in abiotic materials were 0.001 microg L(-1) in rainwater; 0.0015 microg L(-1) in seawater near hydrothermal vents vs. < 0.00004-0.0007 microg L(-1) elsewhere; 5.0 microg kg(-1) dry weight (DW) in the Earth's crust; 19.0 microg L(-1) in a freshwater stream near a gold mining site; 440 microg kg(-1) DW in atmospheric dust near a high traffic road; 843 microg kg(-1) DW in alluvial soil near a Nevada gold mine vs. < 29 microg kg(-1) DW premining; 2.53 mg kg(-1) DW in snow near a Russian smelter vs. < 0.35 mg kg(-1) DW at a reference site; 4.5 mg kg(-1) DW in sewage sludge; 28.7 mg kg(-1) DW in polymetallic sulfides from the ocean floor; and 256.0 mg kg( 1) DW in freshwater sediments near a gold mine tailings pile vs. < 5 microg kg( 1) DW prior to mining. In plants, elevated concentrations of 19 microg Au kg(-1) DW were reported in terrestrial vegetation near gold mining operations vs. < 4 microg kg(-1) DW at a reference site; 37 microg kg(-1) DW in aquatic bryophytes downstream from a gold mine; 150 microg Au kg(-1) DW in leaves of beans grown in soil containing 170 microg kg(-1) DW; up to 1.06 mg kg(-1) DW in algal mats of rivers receiving gold mine wastes; and 0.1-100 mg kg(-1) DW in selected gold accumulator plants. Fish and aquatic invertebrates contained 0.1-38.0 microg Au kg(-1) DW. In humans, gold concentrations up to 1.1 microg L(-1) were documented in urine of dental technicians vs. 0.002-0.85 microg L(-1) in reference populations; 2.1 microg L(-1) in breast milk, attributed to gold dental fillings and jewelry of mothers; 1.4 mg kg(-1) DW in hair of goldsmiths vs. a normal range of 6-880 microg kg(-1) DW; 2.39 mg L(-1) in whole blood of rheumatoid arthritis patients receiving gold thiol drugs to reduce inflammation (chrysotherapy) vs. a normal range of 0.2-2.0 microg L(-1); and 60.0 to 233.0 mg kg(-1) fresh weight (FW) in kidneys of rheumatoid arthritis patients undergoing active chrysotherapy vs. < 42.0 mg kg(-1) FW kidney 140 months posttreatment. PMID- 15887365 TI - Evaluation of coastal waters receiving fish processing waste: Lota Bay as a case study. AB - Liquid wastes from the fish meal and oil processing industries produce serious environmental impacts in coastal embayments on the coasts of Chile and Peru. This article presents an analysis of an environmental monitoring program at Lota Bay, a shallow coastal indentation in central Chile (37 degrees S) exposed to industrial fishing activity. The study of the environmental impact produced by waste effluents permitted making an evaluation of the bay's capacity for seasonal recovery from this impact. Seasonal cruises were carried out during 1994 and in 1996, 1997, and 1998. Variables analyzed included salinity, temperature, dissolved oxygen, ammonium concentration and surface oil and grease. The hydrographic regime of Lota Bay follows a seasonal pattern, where, typical of most SE pacific embayments, waters from subsuperficial oxygen minimum zones moved into the bay. The percentages of dissolved oxygen were critical in the area of organic waste discharge. The impact of wastewater is related to the type and status of the fishery, including: (i) overloads of plant production lines, (ii) maintenance and cleaning of installations, and (iii) degree of shipboard fishing conservation. Major alterations were observed in summer, when the highest discharge of organic load occurred. In winter, an improvement in the re-aeration conditions reduced the impact. Remedial measures implemented beginning in 1997 arose from the monitoring program and had to be separated into two recovery factors including (a) internal management of plants and (b) treatment of plant effluents. PMID- 15887366 TI - Multiple-trait analysis of fluctuating asymmetry levels in anthropogenically and naturally stressed sites: a case study using Chironomus riparius Meigen, 1804 larvae. AB - Fluctuating asymmetry levels were measured in fourth-instar Chironomus riparius larvae collected from various sampling sites in Galicia (northwest Spain) subjected to different types and degrees of stress. Specifically, we selected sites with contamination of anthropogenic origin, ranging from urban and industrial sewages to organic wastes, as well as two sites without chemical contamination, but with marked diurnal variations in their physicochemical conditions. Fluctuating asymmetry levels were determined for various structures of the head capsule (antennal segment I length, antennal segment II length, mentum width, and number of pecten epipharyngis teeth). The results obtained group the sampling sites in a similar way to other measures of stress. In particular, grouping resulted to be similar to that obtained after analysis of head capsule deformities. Moreover, results of our work are consistent with the hypothesis that some characters may possibly be useful indicators of specific types of stress. PMID- 15887367 TI - Augmenting groundwater monitoring networks near landfills with slurry cutoff walls. AB - This study investigated the use of slurry cutoff walls in conjunction with monitoring wells to detect contaminant releases from a solid waste landfill. The 50 m wide by 75 m long landfill was oriented oblique to regional groundwater flow in a shallow sand aquifer. Computer models calculated flow fields and the detection capability of six monitoring networks, four including a 1 m wide by 50 m long cutoff wall at various positions along the landfill's downgradient boundaries and upgradient of the landfill. Wells were positioned to take advantage of convergent flow induced downgradient of the cutoff walls. A five well network with no cutoff wall detected 81% of contaminant plumes originating within the landfill's footprint before they reached a buffer zone boundary located 50 m from the landfill's downgradient corner. By comparison, detection efficiencies of networks augmented with cutoff walls ranged from 81 to 100%. The most efficient network detected 100% of contaminant releases with four wells, with a centrally located, downgradient cutoff wall. In general, cutoff walls increased detection efficiency by delaying transport of contaminant plumes to the buffer zone boundary, thereby allowing them to increase in size, and by inducing convergent flow at downgradient areas, thereby funneling contaminant plumes toward monitoring wells. However, increases in detection efficiency were too small to offset construction costs for cutoff walls. A 100% detection efficiency was also attained by an eight-well network with no cutoff wall, at approximately one-third the cost of the most efficient wall-augmented network. PMID- 15887368 TI - Brazilian LTER: ecosystem and biodiversity information in support of decision making. AB - Brazil officially joined the International Long Term Ecological Research (ILTER) network in January 2000, when nine research sites were created and funded by the Brazilian Council for Science and Technology (CNPq). Two-years later some positive signs already emerge of the scientific, social and political achievements of the Brazilian LTER program. We discuss examples of how ecosystem and biodiversity information gathered within a long-term research approach are currently subsidizing decision-making as regards biodiversity conservation and watershed management at local and regional scales. Success in this respect has often been related to satisfactory communication between scientists, private companies, government and local citizens. Environmental education programs in the LTER sites are playing an important role in social and political integration. Most examples of integration of ecological research to decision-making in Brazil derive from case studies at local or regional scale. Despite the predominance of a bottom-up integrative pathway (from case studies to models; from local to national scale), some top-down initiatives are also in order, such as the construction of a model to estimate the inpact of different macroeconomic policies and growth trajectories on land use. We believe science and society in Brazil will benefit of the coexistence of bottom-up and top-down integrative approaches. PMID- 15887369 TI - Metal leachability and anthropogenic signal in roadside soils estimated from sequential extraction and stable lead isotopes. AB - Several roadside soil samples were collected at two field sites in Sweden. They were analysed for total elemental content (using both ICP-MS and XRF) and stable lead isotopes. Extraction with deicing salt solution and sequential extraction were performed in order to elucidate the potential mobility due to the use of deicing agents. The total concentrations of elements, especially lead, have decreased and lead is presently almost at background concentrations (15-51 ppm for surface samples). However, the isotopic signature indicates that old gasoline lead still is left at the site constructed prior to 1975. The field site constructed in 1992 showed, however, no 206Pb/207Pb ratio below 1.14. Only minor amounts were leached using deicing salt solutions; for lead only 0.29%, on average, was extracted indicating that the mobile fraction already was released. Sequential extraction indicated that lead mainly was associated with reducible (34.4%) and oxidisable (35.4%) fractions. Exchangable and acid soluble fractions contained 20.3% while 10.0% was found in the residual fraction. The salt extraction released, however, very low concentrations indicating that most in fraction 1 is acid soluble (e.g. carbonates). Tungsten was also found at high concentrations indicating a possible impact from studded tires. For tungsten the following composition was obtained: residual (48.0%) > oxidisable (47.6%) > reducible (3.3%) > exchangeable/acid soluble (1.1%). From the isotopic studies it was also suggested that the order for incorporating anthropogenic lead into soils is exchangeable/carbonates > (hydr)oxides > organic matter > residual. The multivariate technique principal component analysis (PCA) seems promising for evaluating large sequential extraction datasets. PMID- 15887370 TI - Environmental degradation and environmental threats in China. AB - The article presents a review of environmental degradation and its threats in China. Air pollution, water pollution, deforestation, soil degradation, sand depositing in dams, decaying urban infrastructure, and more and more hazards such as floods, landslides and soil erosion are major consequences of environmental degradation and are making tremendous loss both in life and property. Through investigation, the author found that poor air quality in the large cities; water pollution in the downstream of many rivers; the multiple problems of many mining areas; lack of access to fresh water; decaying sewage systems; and the disastrous impact of these environmental degradations on public health and agricultural products in many provinces is rather serious. Relationship of environmental degradation and natural hazards is close; more attention should be put in environmental degradation that may surpass economy progress if the trend continues. It is therefore imperative that Chinese government undertake a series of prudent actions now that will enable to be in the best possible position when the current environmental crisis ultimately passes. PMID- 15887371 TI - Multi-objective environmental management in constructed wetlands. AB - We examined multi-objective environmental management as applied to pursuing concurrent goals of water treatment, biodiversity and promotion of recreation in constructed wetlands. A case study of a wetland established to treat landfill leachate, increase biodiversity, and promote recreation was evaluated. The study showed that attempts to combine pollution management with activities promoting biodiversity or recreation are problematic in constructed wetlands. This could be because the typical single-objective focus of scientific research leads to contradictions when planning, implementing and assessing the multi-objective use of wetlands. In the specific case of wetland filters for landfill leachate treatment, biodiversity, and recreation, there is a need for further research that meet practical needs to secure positive outcomes. PMID- 15887372 TI - Metal levels in tissues of Florida gar (Lepisosteus platyrhincus) from Lake Okeechobee. AB - There is considerable concern about the effects of contaminants on organisms at higher trophic levels, as well as interest in developing bioindicators of ecological health. In this article we examine the levels of arsenic, cadmium, chromium, lead, manganese, mercury, and selenium in the gonads, liver and muscle of Florida gar (Lepisosteus platyrhincus) from Lake Okeechobee in south-central Florida. Gar are top-level predators in freshwater ecosystems. We expected that there would be differences in metal levels as a function of tissue, and predicted differences as a function of sex, size, and location around the lake. There were significant differences among tissues for all metals, but there were few differences as a function of size and gender, which may relate to the generally low levels of metals. Mercury levels were significantly higher in gar collected from the north, and arsenic levels were significantly higher in the gar collected from the south. All metal levels in the muscle of gar collected from Lake Okeechobee were similar to, or lower than those generally reported from other areas of the United States. Thus, although the levels of mercury from the Everglades to the south of Lake Okeechobee are sufficiently high to result in human fish consumption advisories, there are none for Lake Okeechobee. The mercury levels in fish muscle from Lake Okeechobee are lower than from many waters in the U.S. and do not pose a problem for humans. However, liver levels average over the 0.5 microg g(-1) level considered safe for human consumption, and might pose a problem for non-human consumers that eat the fish whole. PMID- 15887373 TI - Identification of characteristic regions and representative stations: a study of water quality variables in the Kattegat. AB - Gradients in nutrient distributions and the effects of eutrophication are common features in most coastal marine areas. These structures occur in aquatic systems due to spatial differences in hydrography, nutrient loading and key biogeochemical processes. Two statistical methods, cluster analysis and probability mapping, have been used in the present study to determine characteristics and patterns in water quality variables. Combined, these two methods provide a useful tool to statistically determine spatial homogeneity and representativity of areas and stations. A case study is presented here in which water quality variables (salinity, dissolved inorganic nitrogen, dissolved inorganic phosphorus and chlorophyll) in surface waters of the Kattegat are analysed for the time period 1993-1997. It was found that morphology, the proximity to sources of nutrient loading from land, nutrient uptake and the infrequent water exchange between the Baltic Sea in the south and the Skagerrak in the north all contribute to create distinct regions of water quality. Regions with concentrations significantly different from the overall mean are identified. In addition to identification of regions with similar characteristics, representative stations (as well as not representative stations) for the respective regions were made. This type of information can be used to design new or revise old monitoring programs. PMID- 15887374 TI - Evaluation of water quality projects in the Lake Tahoe basin. AB - Lake Tahoe is a large sub alpine lake located in the Sierra Nevada Range in the states of California and Nevada. The Lake Tahoe watershed is relatively small (800 km(20) and is made up of soils with a very low nutrient content and when combined with the Lake's enormous volume (156 km(3)) produces water of unparalleled clarity. However, urbanization around the Lake during the past 50 yr has greatly increased nutrient flux into the Lake resulting in increased algae production and rapidly declining water clarity. Lake transition from nitrogen limiting to phosphorous limiting during the last 30 yr suggests the onset of cultural eutrophication of Lake Tahoe. Protecting Lake Tahoe's water quality has become a major public concern and much time, effort, and money has been, and will be, spent on this undertaking. The effectiveness of remedial actions is the subject of some debate. Local regulatory agencies have mandated implementation of best management practices (BMPs) to mitigate the effects of development, sometimes at great additional expense for developers and homeowners who question their effectiveness. Conclusive studies on the BMP effectiveness are also expensive and can be difficult to accomplish such that very few such studies have been completed. However, several project evaluations have been completed and more are underway. Such study usually demonstrates support of the project's effectiveness in decreasing nutrient flux to Lake Tahoe. Here, we review the existing state of knowledge of nutrient loading to the Lake and to highlight the need for further evaluative investigations of BMPs in order to improve their performance in present and future regulatory actions. PMID- 15887375 TI - Enhanced wet deposition estimates using modeled precipitation inputs. AB - Effective assessments of linkages between atmospheric deposition and sensitive aquatic and terrestrial ecosystems require a knowledge of spatial patterns in deposition of greater resolution than currently available using point estimates and two-dimensional surface algorithms. A high resolution wet deposition model was developed for the Eastern U.S.A. by incorporating daily precipitation measurements from National Oceanic and Atmospheric Administration (NOAA) monitoring sites located within the Eastern U.S.A.; topographic variables, including elevation, slope, and aspect that affect the amount and distribution of precipitation across the region; and precipitation chemistry data from National Atmospheric Deposition Program (NADP) sites within and adjacent to the region. Model performance, which was tested against independent measurements from three physiographic regions of the eastern U.S.A., revealed a statistically significant reduction in interpolation error compared to traditional two-dimensional surface algorithms. PMID- 15887376 TI - Oceanographic studies off Beypore Port, west coast of India to locate a dredge dumping site. AB - This study presents the salient results of the oceanographic investigations carried out along the west coast of India to locate a dumping site for the dredged material generated from the capital of the proposed development of the all weather port. Based on the results of movement of disposed material obtained from a 2 dimensional coastal circulation model and considering the possibility of having a navigational channel later on in line with the port, a dumping site is recommended at the 15 m water depth contour with coordinates: Latitude 11 degrees 07.00'; Longitude 75 degrees 45.60', where the environmental impacts on the ecosystem due to dumping operations is considered to be minimum. PMID- 15887377 TI - Concentrations of Arsenic, Cadmium, Copper, Lead, Selenium, and zinc in fish from the Mississippi River basin, 1995. AB - Fish were collected in late 1995 from 34 National Contaminant Biomonitoring Program (NCBP) stations and 12 National Water Quality Assessment Program (NAWQA) stations in the Mississippi River basin (MRB), and in late 1996 from a reference site in West Virginia. The NCBP sites represented key points (dams, tributaries, etc.) in the largest rivers of the MRB. The NAWQA sites were typically on smaller rivers and were selected to represent dominant land uses in their watersheds. The West Virginia site, which is in an Eastern U.S. watershed adjacent to the MRB, was selected to document elemental concentrations in fish used for other aspects of a larger study and to provide additional contemporaneous data on background elemental concentrations. At each site four samples, each comprising (nominally) 10 adult common carp (Cyprinus carpio, 'carp') or black bass (Micropterus spp., 'bass') of the same sex, were collected. The whole fish were composited by station, species, and gender for analysis of arsenic (As), lead (Pb), and selenium (Se) by atomic absorption spectroscopy and for cadmium (Cd), copper (Cu), and zinc (Zn) by inductively-coupled plasma emission spectroscopy. Concentrations of most of the elements examined were lower in both carp and bass from the reference site, a small impoundment located in a rural area, than from the NCBP and NAWQA sites on rivers and larger impoundments. In contrast, there were few overall differences between NCBP sites NAWQA sites. The 1995 results generally confirmed the continued weathering and re-distribution of these elemental contaminants in the MRB; concentrations declined or were unchanged from 1984-1986 to 1995 at most NCBP sites, thus continuing two-decade trends. Exceptions were Se at Station 77 (Arkansas R. at John Martin Reservoir, CO), where concentrations have been elevated historically and increased slightly (to 3.8-4.7 microg g-(1) in bass and carp); and Pb, Cd, and Zn at Station 67 (Allegheny R. at Natrona, PA), where levels of these metals were high in the past and increased from 1986 to 1995. PMID- 15887378 TI - Devils advocates. Tasmanian Devil Facial Tumor project. PMID- 15887379 TI - Animal welfare--public policy perspectives. PMID- 15887380 TI - Recent veterinary graduates. Changes in veterinarians' initial career experiences over the last five decades: an abridged report. PMID- 15887381 TI - Over award payments--getting them right and protecting your business from costly claims. PMID- 15887382 TI - A retrospective report of 90 dogs with suspected cane toad (Bufo marinus) toxicity. AB - OBJECTIVE: To report the clinical characteristics of toad toxicity in domestic dogs in Brisbane. DESIGN: A retrospective analysis of clinical cases. PROCEDURE: All cases of toad poisoning which presented to a northern suburbs emergency clinic in Brisbane over a 30-month period beginning in April 1999 were reviewed. RESULTS: A total of 90 canine cases of suspected toad poisoning were reviewed. Small breed dogs accounted for 76% of cases. Jack Russell, Silky, and Fox Terriers were the most represented breeds. Cases were reported year round, with fewest cases over the winter months. The most common clinical signs were increased salivation (78% of cases), and red oral mucous membranes (63% cases). Seizures occurred in 31% of cases. Generally the outcome was excellent with 96% survival. PMID- 15887383 TI - Clostridium septicum arthritis in three foals. PMID- 15887384 TI - Angiotrophic intravascular lymphosarcoma presenting as bi-cavity effusion in a dog. AB - A 9-year-old German Shepherd cross was presented with a history of dyspnoea, decreased activity, episodic collapse and abdominal distention. Cytological examination of thoracic and abdominal fluid demonstrated neoplastic cells, either round or mesenchymal in nature. The dog was found to have a mass in the heart base region. Exploratory thoracotomy was performed to identify the exact nature of the mass histologically, and to determine if it was the source of neoplastic cells seen in the effusions. Histological diagnosis of the mass was mesothelioma. Postoperatively the dog developed thrombocytopenia, probably immune-mediated, which was treated with prednisone therapy. The dog developed signs of respiratory distress shortly before discharge and, due to its deteriorating condition, was euthanased. A necropsy was performed and tissues were collected for histological examination. The histological diagnosis was angiotropic intravascular lymphosarcoma. Diagnosis of mesothelioma was not confirmed. PMID- 15887385 TI - Acute necrotising pancreatitis following grain overload in a donkey. AB - A donkey developed abdominal discomfort and distension together with cardiovascular collapse and gastric reflux 24 to 36 hours after ingesting a large amount of poultry feed. Rectal findings prompted a laparotomy that identified extensive gastric dilation, an empty, atonic small intestine, dry colonic content and an easily corrected caecal displacement. These findings were not consistent with the severity of the signs, which were attributed to endotoxaemia. The donkey was euthanased during recovery. Necropsy revealed acute necrotising pancreatitis with massive gastric dilation and right dorsal colon impaction. The clinicopathological features of acute equine pancreatitis associated with grain overload are discussed. PMID- 15887386 TI - Melioidosis in an alpaca (Lama pacos). PMID- 15887387 TI - Fracture repair in a draught horse. PMID- 15887388 TI - A comparative field trial of cephalonium and cloxacillin for dry cow therapy for mastitis in Australian dairy cows. AB - OBJECTIVE: To investigate and compare the therapeutic efficacy of dry cow agents containing either cephalonium or cloxacillin within Australian dairy herds. DESIGN: A treatment-control trial. METHODS: Milk from infected quarters of cows with high somatic cell counts in milk on eight Australian dairy farms was cultured to identify bacterial pathogens. Cows were randomly assigned to treatment groups and one group was treated with cephalonium at drying off and the other group was treated with cloxacillin at drying off. Milk samples from infected quarters were collected immediately after calving and were cultured for pathogens. The effect of treatment on bacteriological cure was examined and somatic cell counts from infected cows from the first two herd tests after calving were examined for a treatment effect. On four farms, milk samples were collected for culture from all cases of clinical mastitis identified within the first 7 days after calving. The effect of treatment upon incidence of clinical mastitis after calving was examined. RESULTS: There was no significant difference between treatments on quarter cure rates for new infections, for chronic infections and for infections with Staphylococcus aureus, Streptococcus agalactiae and Streptococcus uberis. Infected quarters treated with cephalonium had a significantly higher cure rate than quarters treated with cloxacillin when Corynebacterium bovis and Staphylococcus epidermids were included as pathogens combined (80.3% versus 70.7%). There was no significant difference between the treatments on somatic cell counts of infected cows at the first two herd tests after calving. There was no difference between treatments on the incidence of clinical mastitis in the first 7 days after calving. PMID- 15887389 TI - Nitrate-nitrite toxicity in cattle and sheep grazing Dactyloctenium radulans (button grass) in stockyards. AB - Hungry cattle and sheep introduced to stockyards containing a dominant or pure growth of Dactyloctenium radulans (button grass) suffered acute nitrate-nitrite toxicity in four incidents in inland Queensland between 1993 and 2001. Deaths ranged from 16 to 44%. Methaemoglobinaemia was noted at necropsies in all incidents. An aqueous humour sample from one dead steer contained 75 mg nitrate/L and from one dead sheep contained 100 mg nitrate and 50 mg nitrite/L (normal = ca 5 mg nitrate/L). Both lush and dry button grass were toxic. The nitrate content of button grass from within the stockyards ranged from 4.0 to 12.9% as potassium nitrate equivalent in dry matter and from outside the stockyards ranged from <0.2 to 0.4%. These data suggest that urine and faeces in stockyard soil may boost the nitrate content of button grass to a concentration hazardous to hungry ruminants. PMID- 15887390 TI - Aspergillus clavatus tremorgenic neurotoxicosis in cattle fed sprouted grains. AB - Beef and dairy cattle from four different herds in southern and central Queensland fed hydroponically-produced sprouted barley or wheat grain heavily infested with Aspergillus clavatus developed posterior ataxia with knuckling of fetlocks, muscular tremors and recumbency, but maintained appetite. A few animals variously had reduced milk production, hyperaesthesia, drooling of saliva, hypermetria of hind limbs or muscle spasms. Degeneration of large neurones was seen in the brain stem and spinal cord grey matter. The syndrome was consistent with A clavatus tremorgenic mycotoxicosis of ruminants. The cases are the earliest known to be associated with this fungus in Australia. They highlight a potential hazard of hydroponic fodder production systems, which appear to favour A clavatus growth on sprouted grain, exacerbated in some cases by equipment malfunctions that increase operating temperatures. PMID- 15887391 TI - Mycotoxic nervous disease in cattle fed sprouted barley contaminated with Aspergillus clavatus. PMID- 15887392 TI - Tick paralysis of dogs in Victoria due to Ixodes cornuatus. PMID- 15887393 TI - The role of wild aquatic birds in the epidemiology of avian influenza in Australia. PMID- 15887394 TI - [Contemporary prospects of prevention of type 2 diabetes mellitus]. AB - Prevalence of the Type 2 diabetes mellitus (DM2) has been rising in the whole word. It is assumed that before DM2 develops, patients undergo a stadium of impaired glucose tolerance (IGT) or they have impaired fasting glycaemia (IFG). The confirmed IFG or IGT represent strong predictors of DM2 manifestation and at the same time they are related with high cardiovascular risk, namely with IGT. Other significant risk factor (RF) of DM2 is the obesity and metabolic syndrome. Recent clinical studies have shown that some metabolic abnormalities, which precede development of DM2 can be positively influenced by the lifestyle changes, including improvement of the diet and increasing the physical activity. Such measures can prevent or at least to delay the development of type 2 diabetes mellitus and thus the development of cardiovascular diseases. Positive effect has also the administration of some drugs, already tested in clinical studies, namely glitasons, metromin, inhibitor of ACE, sartans and other. PMID- 15887395 TI - [Conservative therapy of female urinary incontinence--potential and effect]. AB - Non-surgical treatment of female stress urinary incontinence is not as effective as surgical methods but it is very successful in indicated cases. Rehabilitation of the pelvic floor muscles (Kegel exercises, vaginal cones, and electrostimulation of the pelvic floor muscles), drug treatment (alfa-mimetics, tricyclic antidepressives, estrogens, duloxetin), pessarotherapy and uretral obturator devices represent possibilities of conservative therapy of the stress incontinence. Conservative therapy is the method of choice in the treatment of urge incontinence. The most successful are anticholinergic drugs but they have very frequent serious side effects (dryness of the mucous membranes, accommodation disorders, constipation). Spasmolytics, estrogens and tricyclic antidepressives are the other popular used drugs. Life style modification, bladder training and electrostimulation represent very important parts of the conservative treatment. Effectiveness of the non-surgical treatment of both urge and stress urinary incontinence can not reach 100 percent but it helps very much in the quality of life improvement of incontinent women. PMID- 15887396 TI - [Contemporary potential of the mini-invasive surgical treatment of the female stress urinary incontinence]. AB - In 1994 DeLancey published the hammock hypothesis. According to this theory the increase of the urethral closure pressure depends on the urethral compression by the suburethral supportive tissue and the intraabdominal position of the proximal urethra and the bladder neck are not the main factors ensuring the continence. In connection with this new theory Ulmsten published (in 1996) results of the stress incontinence treatment with the tension-free polypropylene vaginal tape (TVT). This operation represents a revolution in the antiincontinent surgery. The success rate of this procedure is about 90% and it is the same in obese and in non obese women. Concomitant TVT and transvaginal procedures for prolapse do not decrease the effectiveness and do not increase the perioperative and postoperative morbidity. The procedure is connected with some peroperative (bladder perforation, retropubic haematoma), early postoperative (urinary retention) and late postoperative complications (urge incontinence, vaginal protrusion of the tape). In 2003 newly developed tape procedure--TOT (transobturator tape) was an evolutionary step in the improvement of tape procedures. The evaluation of the efficacy and complications rate of this modification is premature up to this time but hitherto positive results predetermine this surgical modality to successful expand. PMID- 15887397 TI - [Contemporary view on the plasma natriuretic peptide assessment in the clinical practice]. AB - Plasma natriuretic peptide assessment became a part of the routine clinical practice because of the laboratory methods development. After a brief historical overview of natriuretic peptides discovery, the article focuses on the brain natriuretic peptide (BNP). BNP plays an important role in the pathophysiology of many diseases of cardiovascular system due to its effects on the circulation (natriuresis, diuresis, inhibition of renin-angiotensin system, etc). The authors describe process of BNP synthesis, factors leading to its release into circulation. The main emphasis is done on the clinical significance of BNP and NT proBNP assessment for the-diagnosis in cardiovascular diseases, risk stratification and monitoring of the treatment. PMID- 15887398 TI - [Possibilities of the laboratory diagnostic of metabolic skeletal changes]. AB - Skeletal disorders represent ever-rising problem. Such diseases are of the highly heterogenous origin and for their verification the laboratory diagnostic of metabolic skeletal changes cannot be omitted. Increasing knowledge on the physiology and pathophysiology of the bone tissue metabolism and the use of new laboratory methods can improve the diagnosis and treatment of such critical states. The article gives an overview of contemporary possibilities of the laboratory diagnostic of metabolic skeletal changes. The significance, advantages, disadvantages and possible risks in evaluation of individual osteomarkers are presented. PMID- 15887399 TI - [Foetal ECG in the prediction of intrapartum hypoxia]. AB - Intrapartum foetal hypoxia represents one of the most frequent causes of the hypoxia-ischemia CNS injury in newborns and it can result in the development of a permanent handicap. It often results from the underestimation of the development of the delivery by the obstetrician who conducts delivery and who is responsible for it. That is why the contemporary obstetrics is using new instruments, enabling to evaluate objectively the development of the intrapartum foetal hypoxia. The praxis consequently introduced cardiotocography (CTG), foetal pulse oximetry (FpO2) and recently new methods for evaluation of ST interval in foetal ECG- STAN. The last method has the highest specificity for prediction of the foetal hypoxia and it properly signalises the development of the metabolic foetal acidosis, which threatens the foetus during delivery and which can impair the vital organs. Foetal myocardium sensitively responds to the release of stress hormones, to the development of anaerobic metabolism and to the increase of potassium levels. The development of hypoxia manifests in ECG as a subsequent rise of T wave, elevation of T/QRS segment and as a significantly biphasic ST interval. The last sign indicates serious state of the foetus in utero accompanied with metabolic acidosis. PMID- 15887400 TI - [Promise of immunotherapy of the chronic myeloid leukaemia]. AB - Because of the presence of unique antigens, chronic myeloid leukaemia (CML) represents an appealing target for immunotherapy. The progress achieved in the fields of gene therapy, tumour immunology and vaccinology offers a wide spectrum of methods that could be utilized for the development of therapeutic vaccines against CML. Experience obtained in several clinical studies with peptide-based vaccines have made it clear that it is possible to induce specific immune reactivity; however, its clinical efficacy has been low if any. Studies in mouse systems, which are under way, should be helpful in defining the optimal strategy for immunizing human subjects against bcr-abl positive cells. The author adduces some advantages, but also the limitations, of animal models for this purpose. He also comments on the possibility that the bcr-abl-based therapeutic vaccines might be found ineffective and proposes procedures how to deal with the problem. PMID- 15887401 TI - [Endoscopic investigations in children under 3 years of age at the clinic of paediatric surgery]. AB - BACKGROUND: Retrospective study of gastroscopic investigations in children under 3 years of age. METHODS AND RESULTS: Since 1.1.2002 till 31.10.2004 we performed 119 endoscopic investigations in children of this age. 21 patients were excluded because of missed documentation. The average weight of our children was 11.8 kg. (min. 2.8, max 20 kg). In 55 cases suspicion for the oesophageal or gastric corrosion was the reason of investigations. in 24 cases swallowed alien bodies, in 10 patients we performed dilatation of oesophageal stenosis in repeated sessions in 1-6 moths periods. In 4 children we introduced percutaneous endoscopic gastrostomy (PEG), in 3 children ERCP was performed because of cystic dilatation of the bile duct. In I child the supposed hematemesis was the reason of gastroscopy (negative) and in one patient oesophageal varicula were assumed (negative). CONCLUSIONS: Oesophageal or gastric corrosions were assured in 12 cases (Grade 1.-2.:20.2%). 5 alien bodies couldn't be removed because of peristalsis, 1 object because of its dimensions (25% failures). Our dilations were performed in oesophageal stenosis with bougies or balloon dilator. 9 children were first operated for oesophageal atresia (Vogt IIIb), 1 child was after an operation for oesophageal corrosion with perforation. PMID- 15887402 TI - [Myocardial ischaemia resulting from compression of coronary artery caused by herniation of abdominal organs into thoracic cavity]. AB - A case of 40-year-old patient with compression of coronary artery and myocardial ischaemia is presented. Compression resulted from the herniation of abdominal organs into the thoracic cavity, which developed many years after the car accident trauma. PMID- 15887403 TI - [Shaken baby syndrome]. AB - Shaken baby syndrome represents a specific form of the Abused child syndrome. Injury usually concerns the baby's head and the brain and it is caused by thoughtless treatment accompanied by harsh shaking movements of the head and neck. It can cause a contusion of the cervical spinal hord, a rupture of the bridging veins, intracranial bleeding and a brain tissue impairment either due to the direct axonal damage, or namely by hypoxic-ischaemic insult. The development of the lesion can be fatal, or it can result in permanent impairment of the motor system, in mental or sensory deficits. Occurrence of the syndrome in the Czech Republic is not known, foreign data give 25 cases per 100000 children below one year of age. Injured babies represent over 1% of those hospitalised at paediatric units of intensive care and more than 10% of the death rate at those departments. Proved abuse has forensic consequences: however, convictive evidence can be difficult to obtain. Article gives a concrete case of a boy with the diagnose Shaken baby syndrome. PMID- 15887404 TI - [Psychosomatics is "expensive"]. AB - Experience shoves that number of diseases where recognition and treatment is without limits of classical medicine is rising, however it is fully within the competence of psychosomatic approach. It does not concern the classical classification into organic and functional defects, but it concerns the possibility of complex approach. The theorem of "diagnosis per exclusionem" is still valid, as well as it is true that the means of medicine end at its biological limitations. We consider stressing in our article that psychosomatic diseases or psychosomatic patients do not exist and psychosomatics is not and independent specialization. Psychosomatics is, as inseparable unity of psychic and somatic activities, each human being. Complex biopsychosocial (psychosomatic) approach is the way of thinking and work, which considers human in unrepeatable oneness and context of his life. It does not mean to underestimate objective biological findings and results of instrumentally investigation, but their implanting into the complex network of consequences of the patient's life in order to choose the most appropriate methods of the care of the individual in healthiness and disease. PMID- 15887405 TI - [Patients treated for addictive diseases in the Psychiatric Centre Bohnice in years 1994 to 2003]. AB - During 1994 to 2003 the number of patients treated in the Psychiatric Centre Bohnice who had within the first three dismissory diagnosis alcohol related disorders increased by 77% and disorders caused by other psychoactive drugs increased more than four times. An increase was observed also in pathological gambling (37%). In alcohol related problems the highest increase was seen in drug cessation (about four times). In other drug related problems the highest increase was marked in addiction treatment (almost five times). Such trend probably reflects the situation in the society together with other effects described in the study. PMID- 15887406 TI - [Language and style of texts in the journal of Czech physicians]. AB - This article describes inclination of Czech professional texts in medicine to Anglo-Saxon tradition. Two basic types of articles published in The Journal of Czech Physicians in the period 1990-2002 have been analysed. PMID- 15887407 TI - [Necessity for expert discussion on the state and development of health system- report on the seminar: "Health and Money"]. PMID- 15887408 TI - [Need for health system reform]. PMID- 15887409 TI - [Anticoncept concept based on the patient and insurance company--an essay on the text of the Vice Prime Minister M. Jahn "Health and Money"]. PMID- 15887410 TI - [Clinical neurosciences for efficient treatment--2nd live issue of Clinical Neuroscience/Ideggyogyaszati Szemle]. PMID- 15887411 TI - [Clinico-pathology and differential diagnosis of Binswanger's disease]. AB - Pathologically, Binswanger's disease is subcortical periventricular leucoencephalopathy sparing the U fibers. Clinically it is characterised by executive dysfunction, gait problems, urinary incontinence, pseudobulbar palsy, mood disturbances and dementia. The pathomechanism of Binswanger's disease is unclear. It is hypothesized that it results from an ischemic-hypoxic injury of the periventricular white matter, which, in turn, can be caused by a sclerotic elongation of the medullary arteries, widening of the perivascular spaces or decreased brain perfusion due to hypotension or heart disease. The symptoms of Binswanger's disease frequently overlap with those of normal pressure hydrocephalus, vascular parkinsonism and Alzheimer's disease. A diagnostic criterion of Binswanger's disease is radiologically demonstrated leukoaraiosis, which, on the other hand, is not equivalent with Binswanger's disease. A good clinical response after lumbar puncture or shunt implantation might lead to confusion with normal pressure hydrocephalus, which further complicates the clinical diagnosis. It is likely that among the above mentioned disorders there are a number of transitional forms and overlaps, which might be explained by the common pathomechanism of disturbance in cerebrospinal fluid circulation. PMID- 15887412 TI - [Supplementary sensory-motor seizures--symptomatology, etiology, and surgical management with illustrative case reports]. AB - In the past decade, owing to the advance of epilepsy surgery, growing knowledge has accumulated on the role of the supplementary motor area, described by Penfield and coworkers in the early fifties, in movement regulation and on the characteristics of seizures involving this area. In the Hungarian neurological literature this topic--despite its neurophysiological and practical clinical importance--has been hardly touched. The authors, based on their own experience obtained from surgeries performed within the framework of the "Co-operative Epilepsy Surgery Program", describe the electrophysiological features of this area, its role in movement regulation and the symptoms of epileptic seizures stemmed from or spread onto this area. Using cases as illustrations, they demonstrate the reasoning and various algorithms of the multidisciplinary examination necessary to explore the seizure onset zone and the pathways of seizure spread. Details of the surgical solution are also described. PMID- 15887413 TI - [Primary prevention program of the Hungarian Spine Society--part I. Scientific background of the posture correction exercise scheme]. AB - The primary prevention program of the Hungarian Spine Society aims to increase awareness of the need to develop and automatically maintain a biomechanically correct posture for all school children. The biomechanically correct posture is a dynamic balance based on a correct middle position of the pelvis and on muscle balance. In this position three important anatomical points--the left and right anterior superior iliac spines and the upper medial point of the pubic bone--form one frontal plane. From side-view the imaginary weight median of the body crosses the 2nd to 5th lumbar and the 2nd to 5th cervical vertebral bodies. When the muscles involved in posture are in balance, their strength and flexibility are just appropriate for the almost continuous work required against gravity. In case of static and/or dynamic under- or overload tonic muscles become shortened, and phasic muscles become stretched, and are no longer able to work optimally. Since many muscles and muscle parts that are involved in normal posture maintenance are not satisfactorily challenged in regular physical exercises and sport activities, the preventive exercise scheme of the Hungarian Spine Society aimed to involve these rarely used muscles in special strengthening and stretching exercises. The scheme is based on 12 test exercises that assess the strength and flexibility of postural muscles. A person who is able to do all test exercises correctly has no problem with his or her muscle balance. In order to counteract the harm caused by sedentary lifestyle already in childhood, regular use of this posture correction scheme in physical education starting from preschool throughout the school-years is recommended for all children. PMID- 15887414 TI - [Application of the Multiple Sclerosis Functional Composite in Debrecen]. AB - INTRODUCTION: The Multiple Sclerosis Functional Composite (MSFC) has been recommended by the National Multiple Sclerosis Society as a new clinical outcome measure. It is based on measurements in three clinical dimensions: leg function/ambulation (timed 25-foot walk), arm function (9-hole peg test), and cognitive function (paced auditory serial addition test). Scores on component measures are converted to standard scores (Z-scores) that reflect patient performance. This method has not yet been introduced into routine clinical practice. PATIENTS AND METHOD: MSFC calculation was applied to 17 patients with relapsing-remitting multiple sclerosis (age mean: 37.4+/-10.8 years; duration of the disease: 5.5+/-4.9 years, EDSS: 2.7+/-1.4) seen at the neuroimmunological outpatient clinic to evaluate its usefulness and its correlation with the traditionally applied Expanded Disability Status Scale (EDSS) and with patient reported quality of life. Fifteen patients received immunomodulatory treatment (interferon beta and glatiramer acetate). MSFC and EDSS were measured at 0, 3, 6, 9, 12, 18 months, and questionnaires on quality of life were filled in by the patients at 0, 6, 12, 18 months of follow-up. RESULTS: The prospective study confirmed a strong correlation between EDSS and MSFC (Spearman correlation test, p=0.03, 0.004, 0.002, 0.004, 0.0008, 0.002; R=-0.54, -0.66, 0.68, -0.65, -0.73, 0.69) in multiple sclerosis. The MSFC was more sensitive to clinical changes than EDSS. The extent of changes on the two scales correlated only after 18 months (p<0.005, R=-0.65). The arm/hand function was the most sensitive measure for disease progression. There was no correlation between the quality of life and either of the two other clinical parameters. CONCLUSION: MSFC is a simple method, suitable for follow-up of multiple sclerosis patients in everyday clinical practice. PMID- 15887415 TI - [Volumetric changes following 125I interstitial brachytherapy of gliomas]. AB - BACKGROUND: Image fusion permits quantitative analysis of the consequences of 125 Iodine interstitial irradiation of brain tumors. The volume of tumor necrosis, reactive zone and edema can be compared to the dosimetric data. PATIENTS AND METHOD: Nineteen patients with low grade glioma were analyzed on the average 14.5 months following 125 Iodine interstitial irradiation. Dose planning and image fusion were performed with the Target 1.19 (BrainLab) software. The CT/MR images showing the so called "triple ring" (necrosis, reactive ring and edema) developing after the interstitial irradiation of brain tumors were fused with the planning images and the isodose curves. The volume of the three regions was measured. Values at the intersections of isodose curves and necrosis borders were averaged and used for calculation of tumor necrosis. The volume of normal brain tissue irradiated by given dose values, as well as homogeneity and conformality indices were also determined. RESULTS: The relative volumes of the different parts of the "triple-ring" compared to the reference dose volume were the following: necrosis 54.9%, reactive zone 59.7%, and edema 445.3%. Tumor necrosis developed at 71.9 Gy dose. At the irradiation of an average size glioma with a volume of 12.7 cm3, 5 to 7 cm3 normal brain tissue around the tumor received 60 70 Gy dose. The average homogeneity and conformality indices were 0.24 and 0.57, respectively. CONCLUSION: The analysis of changes in the volume of edema, reactive ring and necrosis caused by interstitial irradiation, and their correlation with the dosimetric data using the image fusion method provide useful information for patient follow-up, clinical management and further therapeutic decisions. PMID- 15887416 TI - [Cognitive theory of psychotrauma and the medical psychotherapy]. PMID- 15887417 TI - ASSA presidential address. No man is an island. ASSA-SAGES Congress, Cape Town, 7 August 2004. PMID- 15887418 TI - The Department of Surgery: University of Pretoria. PMID- 15887419 TI - Modern trends in oesophageal surgery. PMID- 15887420 TI - Management of sigmoid volvulus in Polokwane-Mankweng Hospital. AB - OBJECTIVE: To evaluate the outcome of treatment of patients with sigmoid volvulus in the Polokwane-Mankweng Hospital and to identify the best management options for these patients. METHODS: A retrospective study was undertaken of 85 patients with sigmoid volvulus treated in Polokwane-Mankweng Hospital during the period July 1997-May 2004. RESULTS: In total, 85 patients were evaluated (77 males and 8 females, male/female ratio 9:1). The age range was 7-80 years (mean 42 years). Sigmoidoscopic derotation was attempted in 17 patients, and was successful in 10 patients. Laparotomy was done in 84 patients, viz. 75 emergencies and 9 electives. During laparotomy, gangrenous sigmoid colon was found in 30 patients and viable sigmoid in 54 patients. Resection with primary anastomosis was done in 44 patients. Hartmann's procedure was performed in 33 patients. Sigmoidopexy was done in 7 patients. Total hospital mortality was 6% (5 deaths). Mortality in the 84 operated cases was 5% (4 deaths). CONCLUSIONS: There was no mortality in patients undergoing elective resection and primary anastomosis after successful preoperative deflation and in patients with viable sigmoid volvulus who underwent an emergency Hartmann's procedure. There was low mortality in those patients with resection and primary anastomosis on viable sigmoid (3%, 1:39). The highest mortality (1:5) occurred in cases of resection and primary anastomosis of gangrenous sigmoid colon. PMID- 15887421 TI - Blunt injury of the infrarenal inferior vena cava--imaging and conservative management. AB - Isolated rupture of the infrarenal segment of the inferior vena cava due to blunt trauma is relatively rare. It may be missed clinically and even diagnostic peritoneal lavage may prove negative. The mainstay of diagnosis remains a high degree of clinical suspicion together with sectional imaging. If the patient is haemodynamically stable after initial resuscitation and there is no other indication for exploratory surgery, these lesions can be managed conservatively with good outcome as shown in the case described in this report. PMID- 15887422 TI - Fluctuating pulse deficits associated with intimal arterial injury following gunshot wounds of the extremity--a sign not to be missed. AB - Gunshot wounds may result in intimal arterial injury without breach of the arterial wall integrity. Haemorrhage, haematoma and a pulse deficit are therefore not always found. We report on two cases of lower extremity gunshot wounds with temporal variations in the clinical and radiological assessment of the pedal pulses. In both cases surgical exploration revealed intact arterial vessel walls but significant intimal injury with overlying thrombus. We propose that the pulse deficits were due to distal thromboembolism. Subsequent clot lysis led to a return of the original pulse deficit. Variation in the distal pulses in patients with gunshot wounds of the extremities should alert one of the possibility of an intimal arterial injury; imaging of the vessels is therefore advised. PMID- 15887423 TI - Frederick banting--Nobel laureate for discovery of insulin. PMID- 15887424 TI - Getting there: statin plus ezetimibe for low-density lipoprotein cholesterol goals. PMID- 15887425 TI - A community-based, randomized trial of ezetimibe added to statin therapy to attain NCEP ATP III goals for LDL cholesterol in hypercholesterolemic patients: the ezetimibe add-on to statin for effectiveness (EASE) trial. AB - OBJECTIVE: To determine the extent of reduction in low-density lipoprotein cholesterol (LDL-C) level and improvement in National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) LDL-C goal attainment when ezetimibe was added to ongoing statin therapy in a diverse population of community-based patients. PATIENTS AND METHODS: In this multicenter, double blind, placebo-controlled trial (from January 2003 to August 2003), hypercholesterolemic patients (from 299 US primary care and specialty practices) with LDL-C levels exceeding NCEP ATP III goals were randomized (2:1) to receive ezetimibe (10 mg/d) or placebo in addition to their ongoing statin therapy for 6 weeks. RESULTS: In a study of 3030 randomized patients, ezetimibe added to statin therapy significantly reduced the LDL-C level by an additional 25.8% in the total population, compared with an additional 2.7% reduction with placebo plus statin (treatment difference, -23.1%; P<.001); the treatment difference ranged from 19.9% to -24.0% (P<.001) in each NCEP ATP III risk category subgroup. Significantly (P<.001) more patients (71.0%) treated with ezetimibe added to statin reached their NCEP ATP III target LDL-C level compared with those treated with placebo plus statin (20.6%). The addition of ezetimibe also resulted in improvement in other lipid parameters and high-sensitivity C-reactive protein levels. These benefits were consistent across sex, race, age, statin brand, and dose subgroups. Ezetimibe plus statin therapy was well tolerated, with a safety profile similar to placebo plus statin. CONCLUSION: Across multiple subgroups, ezetimibe added to statin therapy consistently produced significant additional improvements in LDL-C levels and goal attainment, as well as in other lipoproteins, compared with addition of placebo. The addition of ezetimibe to statin therapy should be considered for patients not achieving their NCEP ATP III LDL-C goals while receiving statin therapy alone. PMID- 15887426 TI - Pathogenesis of unexplained drowning: new insights from a molecular autopsy. AB - OBJECTIVE: To perform a molecular autopsy involving the RyR2-encoded cardiac ryanodine receptor/calcium release channel to determine whether mutations responsible for catecholaminergic polymorphic ventricular tachycardia (CPVT) represent a novel pathogenic basis for unexplained drownings. METHODS: A cardiac channel molecular autopsy was performed on 2 individuals who died of unexplained drowning and whose cases were referred to the Sudden Death Genomics Laboratory at the Mayo Clinic in Rochester, Minn. Comprehensive mutational analysis of all 60 protein-encoded exons of the 5 long QT syndrome-causing cardiac channel genes and a targeted analysis of 18 RyR2 exons known to host RyR2-mediated CPVT-causing mutations (CPVT1) was performed using polymerase chain reaction, denaturing high performance liquid chromatography, and DNA sequencing. RESULTS: Both individuals harbored novel mutations in RyR2. Postmortem mutational analysis revealed a familial missense mutation in exon 14, R414C, in a 16-year-old girl. A 9-year-old boy possessed a sporadic missense mutation in exon 49, V2475F. Both amino acid positions involve highly conserved residues that localize to critical functional domains in the calcium release channel. Neither substitution was present in 1000 reference alleles. CONCLUSIONS: This molecular autopsy study provides proof of principle that RyR2 mutations can underlie some unexplained drownings. A population-based genetic epidemiology study that involves molecular autopsies of individuals who die of unexplained drowning is needed to determine the prevalence and spectrum of KCNQ1 and now RyR2 mutations as potential pathogenic mechanisms for drowning. PMID- 15887427 TI - Immunogenicity and reactogenicity of a novel vaccine for human papillomavirus 16: a 2-year randomized controlled clinical trial. AB - OBJECTIVE: To evaluate the immunogenicity, reactogenicity, and tolerability of a prototype human papillomavirus (HPV) 16 viruslike particle (VLP) vaccine directed against the L1 capsid protein. SUBJECTS AND METHODS: We enrolled healthy nonpregnant women aged 18 to 26 years into a 2-year, double-blind, dose-ranging multicenter trial (October 12, 1998, to September 30, 2001). Subjects were assigned to study groups to receive a 3-dose regimen (day 0, month 2, and month 6) of 1 of 4 vaccine doses: 10 microg, 20 microg, 40 microg, or 80 microg or placebo. Serum anti-HPV 16 L1 antibody (sL1Ab) geometric mean titers (GMTs) were measured at day 0, at month 3, at month 7, and every 6 months for a total of 2 years using a radioimmunoassay. The primary immunogenicity analyses evaluated GMTs at month 7 in L1Ab-seronegative subjects at baseline. Vaccine tolerability was also assessed. RESULTS: A total of 480 subjects were randomized to receive placebo (n=52) or 10 microg (n=112), 20 microg (n=105), 40 microg (n=104), or 80 microg (n=107) of HPV 16 L1 VLP vaccine. At baseline, 75% of subjects were L1Ab seronegative. All vaccine doses produced a statistically significant sL1Ab response vs placebo (P<.001). At the completion of the vaccination regimen, sL1Ab GMTs in baseline-seronegative subjects were 36- to 78-fold higher than the sL1Ab GMT at day 0 observed in subjects who had mounted an immune response to HPV 16 infection before enrollment. Serum L1Ab GMTs remained high throughout the 1.5 year postvaccination period. Postvaccination sL1Ab GMTs were 1.1- to 2.4-fold higher in women who had detectable sL1Ab levels at enrollment compared with those in baseline-seronegative subjects, particularly in the persistence phase. The vaccine was generally well tolerated with no statistically significant differences in injection site or systemic adverse experiences among treatment groups. CONCLUSION: Immunization with this novel HPV 16 L1 VLP vaccine was well tolerated and produced an immunogenic response that persisted for at least 1.5 years after the final dose. PMID- 15887428 TI - Postural pseudoanemia: posture-dependent change in hematocrit. AB - OBJECTIVE: To determine the magnitude of posture-related changes in blood components. SUBJECTS AND METHODS: Twenty-eight healthy subjects were studied between 1995 and 2004 at the Vanderbilt Autonomic Dysfunction Center, Nashville, Tenn. Lying and standing plasma volume (PV) and hematocrit (Hct) values were determined for each subject. RESULTS: Individual PV decreases on standing ranged from 6% to 25%. The absolute mean +/- SD PV shift was 417+/-137 mL (range, 149 717 mL). The mean +/- SD change in Hct was from 37.7%+/-2.8% while supine to 41.8%+/-3.2% within 30 minutes of standing. This absolute increase in Hct of 4.1%+/-1.3% represents a relative increase of 11.0%+/-3.6% from lying to standing. CONCLUSIONS: Changes in posture can lead to substantial changes in Hct, which may be attributed mistakenly to blood loss or acute anemia and result in a cascade of unnecessary diagnostic costs. In reality, these changes represent postural pseudoanemia, a normal physiological response to a change in position from standing to lying (and vice versa). PMID- 15887429 TI - Disclosure of natural product use to primary care physicians: a cross-sectional survey of naturopathic clinic attendees. AB - OBJECTIVE: To determine factors that predict disclosure of natural product use by patients to their primary care physicians. METHODS: An 18-item survey about health care use and communication was distributed to patients who presented to the Canadian College of Naturopathic Medicine in March 2003. We examined correlations between responses and created a multivariable logistic regression model to explore which factors were associated with patients' discussion of natural product use with their primary care physicians. RESULTS: The survey was given to 198 patients, 174 of whom responded (response rate, 87.9%). Most respondents (161 [92.5%] of 174) made use of natural products, and many (61 [41.5%] of 147) reported that they did not discuss natural product use with their primary care physician. The most significant factor predictive of patients having discussed natural product use with their primary care physician was having their primary care physician ask about natural product use (odds ratio, 18.77; 95% confidence interval, 5.06-69.62). Most respondents (107 [69.9%] of 153) indicated that their primary care physician did not ask about natural product use. CONCLUSIONS: An important number of patients who visit a naturopathic clinic do not disclose natural product use to their primary care physicians. Our findings suggest that disclosure rates could be improved by physicians taking a more active role in asking their patients about natural product use. PMID- 15887430 TI - Laparoscopy for the definitive diagnosis of intra-abdominal lymphoma. AB - OBJECTIVE: To assess the efficacy of laparoscopy in the diagnosis of intra abdominal lymphoma. PATIENTS AND METHODS: The medical records of patients with suspected primary or recurrent lymphoma who underwent laparoscopy between March 1991 and March 2003 were reviewed. Demographic, clinical, operative, and pathologic data were collected. The feasibility, safety, and effectiveness of the laparoscopic procedure were assessed. RESULTS: Laparoscopic biopsy was attempted in 94 patients. In 78 patients (83%), the procedure was completed laparoscopically. Conversion to laparotomy was undertaken in 16 patients (17%), most commonly because of inadequate exposure, insufficient tissue, or postoperative adhesions. Among the 69 cases of lymphoma, 55 (80%) were diagnosed via laparoscopy only, 9 (13%) via laparotomy, and 5 (7%) with later procedures. Of the remaining 25 patients, 7 had nonlymphoma disease (4 occult carcinomas, 1 multiple myeloma, 1 epithelioid leiomyosarcoma, and 1 neuroblastoma), and 18 had benign lymphadenopathy (no evidence of lymphoma with a mean follow-up of 53 months). The laparoscopic procedure resulted in false-negative results in 6 patients (6%). The mean hospital stay for patients having a laparotomy was 6 days (range, 3-10 days); the remaining patients were all outpatients. The only Intraoperative laparoscopic complication was hemorrhage that required laparotomy. This event occurred in a patient with a previously undiagnosed neuroblastoma. CONCLUSION: Laparoscopic lymph node biopsy safely provides adequate tissue for full histological evaluation on an outpatient basis in most patients with intra abdominal lymphoma. PMID- 15887431 TI - Fifty years of open heart surgery at the Mayo Clinic. PMID- 15887432 TI - Heredofamilial brain calcinosis syndrome. AB - Brain calcinosis syndrome (BCS) usually is defined as bilateral calcium accumulation in the brain parenchyma, primarily in the basal ganglia. More than 50 reported clinical conditions have been associated with BCS. We reviewed clinical, radiological, and genetic features of heredofamilial BCS accompanying all conditions associated with calcium accumulation in the brain reported in English between 1962 and 2003 in MEDLINE. The location, extent, and degree of calcification in the brain show diversity not only among the various disorders but also among patients sharing the same condition. The pathogenesis of BCS is uncertain. More complicated mechanisms may be Involved when brain calcinosis is present but calcium, phosphorus, and parathyroid hormone metabolism abnormalities are absent. We review conditions associated with heredofamilial BCS in which brain calcinosis is nearly uniformly present because such information may be Important to the clinician pursuing an investigative strategy. PMID- 15887433 TI - Nicotine replacement therapy and cardiovascular disease. AB - Smoking is a well-established and important risk factor for cardiovascular disease. Cessation of smoking clearly decreases the chances of a first or subsequent cardiovascular event. Nicotine replacement therapy (NRT) is a proven adJunctive therapy to increase the probability of quitting smoking. Anecdotal reports of adverse events in patients using NRT have led some to question its safety. Is nicotine, whether in tobacco products or in NRT, the cause of the cardiovascular consequences associated with tobacco use? Is using NRT to assist with smoking cessation safer than smoking? Should health care professionals avoid recommending NRT for patients with established cardiovascular disease? This article summarizes the mechanisms of harm associated with smoking and reviews the safety of NRT in both the general population and the population with cardiovascular disease. Recommendations for NRT use are offered. PMID- 15887434 TI - Hyperhidrosis: evolving therapies for a well-established phenomenon. AB - The socially embarrassing disorder of excessive sweating, or hyperhidrosis, and its treatment options are gaining widespread attention. In order of frequency, palmar-plantar, palmar-axillary, Isolated axillary, and cranlofacial hyperhidrosis are distinct disorders of sudomotor regulation. A common link among these disorders is an excessive, nonthermoregulatory sweat response often to emotional stimuli in body regions influenced by the anterior cingulate cortex as opposed to the thermoregulatory sweat response regulated by the preoptic-anterior hypothalamus. Diagnosis of these mechanistically ambiguous disorders is primarily from patient history and physical examination, whereas results of laboratory studies performed with indicator powder reveal the distribution and severity of resting hyperhidrosis and document the integrity of thermoregulatory sweating. Treatment options lie on a continuum based on the severity of hyperhidrosis and the risks and benefits of therapy. In general, therapy begins with antiperspirants or anticholinergics. Iontophoresis is available for palmar plantar and axillary hyperhidrosis. Botulinum toxin type A or local excision/curettage is effective for isolated axillary hyperhidrosis not responsive to topical application of aluminum chloride. Endoscopic thoracic sympathectomy may be used for severe cases of palmar-plantar and palmar-axillary hyperhidrosis. No sole therapy of choice has emerged for craniofacial sweating. The long-term sequelae of hyperhidrosis and its treatment also are discussed. PMID- 15887435 TI - 36-Year-old woman with loss of consciousness, fever, and tachycardia. PMID- 15887436 TI - Cocaine-induced pseudovasculitis. AB - Pseudovasculitis is a disease process that mimics the presentation and possibly the laboratory findings of true vasculitis. However, biopsy specimens do not reveal the typical histopathologic findings expected in vasculitis. One often overlooked cause of pseudovasculitis is cocaine use, which has been described in case reports to cause aggressive nasal destruction and various skin lesions and thus has been confused with Wegener granulomatosis or leukocytoclastic vasculitis. Unfortunately, serologic tests such as antinuclear antibody or antineutrophil cytoplasmic antibody cannot reliably differentiate between these entities. We describe a patient who presented with what was believed to be Wegener granulomatosis affecting the skin and upper airway. However, findings from repeated biopsies did not support this diagnosis, and the only unifying diagnosis was cocaine-induced pseudovasculitis. The ability to recognize and differentiate between true vasculitis and pseudovasculitis is essential for the clinician because treatment options are radically disparate. PMID- 15887437 TI - Cardiomyopathy associated with celiac disease. AB - Celiac disease or celiac sprue is predominantly a disease of the small intestine characterized by chronic malabsorption in genetically susceptible individuals who ingest grains containing gluten, such as wheat, barley, and rye. Although previously believed to be uncommon, celiac disease may be present in up to 1% of the general population. Celiac disease is associated frequently with iron deficiency anemia, dermatitis herpetiformis, selective IgA deficiency, thyroid disorders, diabetes mellitus, and various connective tissue disorders but is rarely associated with cardiomyopathy. We describe a patient with celiac disease associated with cardiomyopathy whose cardiac function improved substantially after treatment with a gluten-free diet. Cardiomyopathy associated with celiac disease is a serious and potentially lethal condition. However, with early diagnosis and treatment with a gluten-free diet, cardiomyopathy in patients with celiac disease may be completely reversible. PMID- 15887438 TI - Catamenial pneumothorax. AB - Catamenial pneumothorax is defined as spontaneous pneumothorax occurring within 72 hours before or after onset of menstruation. Although catamenial pneumothorax is the most common clinical manifestation of intrathoracic endometriosis, this latter condition is not universally identified in women with catamenial pneumothorax and cannot fully explain the recurrent and cyclical episodes of pneumothorax. Therefore, the etiology of this syndrome is unknown, although many theories have been proposed to explain it. We describe a 37-year-old woman with recurrent episodes of spontaneous right-sided pneumothorax and chest pain that occurred close to her menstrual periods. The patient's condition did not abate after initial surgical exploration with abrasive mechanical pleurodesis or after hormonal suppressive therapy at an institution elsewhere. The patient was referred to our institution for further evaluation. A second surgical inspection of the pleural cavity and diaphragm disclosed the presence of multiple diaphragmatic fenestrations that were closed surgically at that time. Postoperatively, the patient discontinued hormonal suppressive therapy, and menstrual cycles became regular. Six months after surgery, the patient remains asymptomatic with no evidence of recurrence of pneumothorax. This case supports recent reports that diaphragmatic defects are often present in patients with catamenial pneumothorax. Surgical exploration to inspect the diaphragm and to close all Identified defects should be performed in patients who continue to experience pneumothorax despite effective hormonal suppression. PMID- 15887439 TI - Genetic testing in the myelodysplastic syndromes: molecular insights into hematologic diversity. AB - The myelodysplastic syndromes (MDS) are associated with a diverse set of acquired somatic genetic abnormalities. Bone marrow karyotyping provides important diagnostic and prognostic information and should be attempted in all patients who are suspected of having MDS. Fluorescent in situ hybridization (FISH) studies on blood or marrow may also be valuable in selected cases, such as patients who may have 5q- syndrome or those who have undergone hematopoletic stem cell transplantation. The MDS-associated cytogenetic abnormalities that have been defined by karyotyping and FISH studies have already contributed substantially to our current understanding of the biology of malignant myeloid disorders, but the pathobiological meaning of common, recurrent chromosomal lesions such as del(5q), del(20q), and monosomy 7 is still unknown. The great diversity of the cytogenetic findings described in MDS highlights the molecular heterogeneity of this cluster of diseases. We review the common and pathophysiologically interesting genetic abnormalities associated with MDS, focusing on the clinical utility of conventional cytogenetic assays and selected FISH studies. In addition, we discuss a series of well-defined MDS-associated point mutations and outline the potential for further insights from newer techniques such as global gene expression profiling and array-based comparative genomic hybridization. PMID- 15887440 TI - Gallstone ileus. PMID- 15887441 TI - Acute sin nombre hantavirus infection complicated by renal failure requiring hemodialysis. PMID- 15887442 TI - Use of the Internet for medication purchases and health information: can we make it safer? PMID- 15887443 TI - Prevention of systemic embolization associated with treatment of gastric fundal varices. PMID- 15887444 TI - Should asymptomatic bacterial vaginosis be treated? PMID- 15887445 TI - Marjolin's ulcer in chronic hidradenitis suppurativa. PMID- 15887446 TI - Hyperglycemic crises: improving prevention and management. PMID- 15887447 TI - Anticonvulsant medications for migraine prevention. PMID- 15887448 TI - Is pharmacotherapy useful in social phobia? PMID- 15887449 TI - Diabetic ketoacidosis. AB - A diagnosis of diabetic ketoacidosis requires the patient's plasma glucose concentration to be above 250 mg per dL (although it usually is much higher), the pH level to be less than 7.30, and the bicarbonate level to be 18 mEq per L or less. Beta-hydroxybutyrate is a better measurement of the degree of ketosis than serum ketones. Intravenous insulin and fluid replacement are the mainstays of therapy, with careful monitoring of potassium levels. Phosphorous and magnesium also may need to be replaced. Bicarbonate therapy rarely is needed. Infection, insulin omission, and other problems that may have precipitated ketoacidosis should be treated. Myocardial infarction is a precipitating cause of diabetic ketoacidosis that is especially important to look for in older patients with diabetes. Cerebral edema is a major complication that occurs primarily in children. Education to prevent recurrence should be offered to all patients, including how to manage sick days and when to call a physician. PMID- 15887450 TI - Diabetic ketoacidosis: what it is and how to prevent it. PMID- 15887451 TI - Hyperosmolar hyperglycemic state. AB - Hyperosmolar hyperglycemic state is a life-threatening emergency manifested by marked elevation of blood glucose, hyperosmolarity, and little or no ketosis. With the dramatic increase in the prevalence of type 2 diabetes and the aging population, this condition may be encountered more frequently by family physicians in the future. Although the precipitating causes are numerous, underlying infections are the most common. Other causes include certain medications, non-compliance, undiagnosed diabetes, substance abuse, and coexisting disease. Physical findings of hyperosmolar hyperglycemic state include those associated with profound dehydration and various neurologic symptoms such as coma. The first step of treatment involves careful monitoring of the patient and laboratory values. Vigorous correction of dehydration with the use of normal saline is critical, requiring an average of 9 L in 48 hours. After urine output has been established, potassium replacement should begin. Once fluid replacement has been initiated, insulin should be given as an initial bolus of 0.15 U per kg intravenously, followed by a drip of 0.1 U per kg per hour until the blood glucose level falls to between 250 and 300 mg per dL. Identification and treatment of the underlying and precipitating causes are necessary. It is important to monitor the patient for complications such as vascular occlusions (e.g., mesenteric artery occlusion, myocardial infarction, low-flow syndrome, and disseminated intravascular coagulopathy) and rhabdomyolysis. Finally, physicians should focus on preventing future episodes using patient education and instruction in self-monitoring. PMID- 15887452 TI - Evaluation of palpable breast masses. AB - Palpable breast masses are common and usually benign, but efficient evaluation and prompt diagnosis are necessary to rule out malignancy. A thorough clinical breast examination, imaging, and tissue sampling are needed for a definitive diagnosis. Fine-needle aspiration is fast, inexpensive, and accurate, and it can differentiate solid and cystic masses. However, physicians must have adequate training to perform this procedure. Mammography screens for occult malignancy in the same and contralateral breast and can detect malignant lesions in older women; it is less sensitive in women younger than 40 years. Ultrasonography can detect cystic masses, which are common, and may be used to guide biopsy techniques. Tissue specimens obtained with core-needle biopsy allow histologic diagnosis, hormone-receptor testing, and differentiation between in situ and invasive disease. Core-needle biopsy is more invasive than fine-needle aspiration, requires more training and experience, and frequently requires imaging guidance. After the clinical breast examination is performed, the evaluation depends largely on the patient's age and examination characteristics, and the physician's experience in performing fine-needle aspiration. PMID- 15887453 TI - Initial evaluation of the patient with suspected dementia. AB - Dementia is a common disorder among older persons, and projections indicate that the number of patients with dementia in the United States will continue to grow. Alzheimer's disease and vascular dementia account for the majority of cases of dementia. After a thorough history and physical examination, including a discussion with other family members, a baseline measurement of cognitive function should be obtained. The Mini-Mental State Examination is the most commonly used instrument to document cognitive impairment. Initial laboratory evaluation includes tests for thyroid-stimulating hormone and vitamin B12 levels. Structural neuroimaging with noncontrast computed tomography or magnetic resonance imaging also is recommended. Other testing should be guided by the history and physical examination. Neuropsychologic testing can help determine the extent of cognitive impairment, but it is not recommended on a routine basis. Neuropsychologic testing may be most helpful in situations where screening tests are normal or equivocal, but there remains a high level of concern that the person may be cognitively impaired. PMID- 15887454 TI - Fibroids (uterine myomatosis, leiomyomas). PMID- 15887455 TI - Management of subclinical hypothyroidism. PMID- 15887456 TI - "My rings won't fit anymore". Ectopic growth hormone-secreting tumor. PMID- 15887457 TI - HIV testing on demand. PMID- 15887458 TI - The treatment of children. Are we prepared to meet tomorrow's challenges? PMID- 15887459 TI - Billing errors can be costly. PMID- 15887460 TI - Radiograph mix-up. PMID- 15887461 TI - What a dilemma. PMID- 15887462 TI - Give 'til it hurts. PMID- 15887463 TI - The damn millimeter. PMID- 15887464 TI - Who's in charge here? PMID- 15887465 TI - Dental caries: an infectious and transmissible disease where have we been and where are we going? AB - Dental caries is often referred to as an infectious disease, which means different things to different people. Pediatric dentists may be tempted to liken caries to other childhood diseases they encounter yet treat caries from mostly a mechanical or surgical approach. On the other end of the spectrum are clinicians who view the oral cavity as a site teaming with bacteria and think that reducing the overall microbial mass using antimicrobial agents will somehow affect this infectious disease. Here, we argue for another view, that is, to combine the most effective portion of each therapy to yield an effective outcome: the treatment and prevention of dental caries. Because primary teeth are eventually shed, pediatric dentists may be in an advantageous position to implement this approach since restorations to primary teeth need not be as durable as those made to permanent teeth. Moreover, pediatric dentists use sealants extensively and are used to working, for example, with rubber dams, especially in young patients. It may turn out that after more research is completed on the ontogeny of the oral microbial flora, changes or ecological shifts made in childhood become lifelong. More specifically, drastic reductions or eliminations of cariogenic bacteria, along with their sites for colonization in the primary dentition, may prevent or delay their transfer to the permanent dentition. Similarly, reductions in a mother's levels of cariogenic bacteria may delay or interrupt the transfer of cariogenic bacteria to her infant. Here, the goal is to discuss several concepts before their final inception into practices and to stimulate discussion of ideas, reframing the design to meet the demands of everyday practice. Ultimately, the proof will come from a large clinical trial conducted in many practices and patient populations. PMID- 15887466 TI - Oral candidiasis in pediatric HIV patients. AB - Oral candidiasis can be an early sign of illness or disease progression in HIV/AIDS and other immuno-compromised states. Oral lesions associated with fungal infections present in a variety of forms, including a few of previously unknown etiology. Diagnosing these variants of disease can be challenging because of their atypical clinical presentation. Moreover, the emergence of new candida species, drug resistance and immature immune systems add to the complexity of this condition, especially in children. PMID- 15887467 TI - Contemporary space maintenance for the pediatric patient. AB - Primary teeth play a critical role in the growth and development of a child. In addition to their role in esthetics, eating, speech, and to encourage normal function and resultant expected growth, the other main function of a primary tooth is to hold space for the permanent successor until it is ready to erupt. Space maintenance forms an integral part of preventive and interceptive orthodontics. After the premature loss of a tooth, not only do space maintainers maintain function and preserve arch length, they also maintain esthetics and eliminate any potential psychological damage a child could face as a result of the premature loss of teeth. The space maintainer also allows the permanent tooth to erupt unhindered into proper alignment and occlusion. PMID- 15887468 TI - Root fracture. Report of case supporting decreased splint times. AB - This is a case presentation of a root fracture that was treated and followed intermittently for 23 years. The treatment outcome supports the newly evolving protocol that root fractures are most successfully treated using a flexible splint over a relatively short duration. PMID- 15887469 TI - Cardiovascular function and basics of physiology in microgravity. AB - Space exploration is a dream of mankind. However, this intriguing environment is not without risks. Life, and the human body, has developed all over evolution in the constant presence of gravity, especially from the moment on when living creatures left the ocean. When this gravitational force is no longer acting on the body, drastic changes occur. Some of these changes occur immediately, others progress only slowly. In the past 40 years of human space flight (first orbital flight by Yuri Gagarin on 12 April, 1961) several hazards for the human body have been identified. Bone mineral density is lost, muscle atrophy and cardiovascular deconditioning occur; pulmonary function, fluid regulating systems of the body, the sensory and the balance system are all disturbed by the lack of gravity. These changes in human physiology have to be reversed again when astronauts return to earth. This can cause adaptation problems, especially after long duration space flights. Also the reaction of human physiology to radiation in space poses a huge risk at this moment. In this review the accent will be on cardiovascular function in space: how normal function is modified to reach a new equilibrium in space after short- and long-duration exposure to microgravity. In order to make long-duration space flight possible the mechanisms of this physiological adaptation must be understood to full extent. Only with this knowledge, effective countermeasures can be developed. PMID- 15887470 TI - Angiotensin-converting enzyme polymorphism in healthy young subjects: relationship to left ventricular mass and functions. AB - OBJECTIVES: Angiotensin-converting enzyme (ACE) is a key enzyme in the production of angiotensin II and thus may participate in the modulation of cardiac growth. The cloning of the ACE gene has made it possible to identify a deletion (D) insertion (I) polymorphism that appears to affect the level of serum ACE activity. The aim of the study is to analyse the ACE gene I/D polymorphisms in healthy young subjects and to evaluate its relationship to left ventricular mass and functions. METHODS: 38 women and 40 men (mean age 21.1 +/- 1.7 and 21.4 +/- 1.7 years) were studied. They underwent complete echocardiographic assessment and analysis of ACE insertion (I) and deletion (D) allele frequencies in peripheral blood by polymerase chain reaction. Thickness of interventricular septum (IVS) and posterior wall (LVPW) and left ventricular mass (LVM) and LVM index (LVMI) were measured by M-mode. Left ventricular ejection fraction (LVEF) was calculated by Simpson's method. RESULTS: There was no statistically significant difference among the DD, DI and II genotypes, concerning age, body mass index, heart rate, systolic and diastolic blood pressures.The thickness of IVS (9.5 mm), LVPW (9.0 mm), LVM (204.5 g) and LVMI (105.5 g/m2) in DD genotypes were higher than both DI (8.3 mm; 8.1 mm; 168.1 g; 90.9 g/m2) and II genotypes (8.2 mm; 7.0 mm; 141.7 g; 77.8 g/m2) in men, but not in women. LVEF among the 3 genotypes were not statistically different. CONCLUSION: Our findings suggest that left ventricular hypertrophy is partially determined by genetic disposition especially in men but not in women. PMID- 15887471 TI - Simvastatin: pharmacological response in experimental hyperfibrinogenaemias. AB - Through a disorder in the endothelial haemostatic balance, hyperfibrinogenaemia could generate endothelial dysfunction. Statins would have antiinflammatory effects on injured endothelium. OBJECTIVE: Simvastatin pharmacological response in rats with hyperfibrinogenaemias induced by laparotomies was studied. METHODS AND RESULTS: Rats were subjected to multiple injuries (MI) for 30 days (1 laparotomy/week) and for 60 days (1 laparotomy/2 weeks). Simvastatin (0.035 mg/kg) was administered orally to the 30-day multiple injuries group after the third injury for a period of 10 days. A similar dose was administered to the 60 day multiple injuries group after the second injury for a period of 45 days. Blood samples of all the groups were obtained 72 hours after the last injury. In the 30 and 60-day multiple injuries groups, a statistically significant fibrinogen increase was observed (336.6 +/- 7.5 and 358.7 +/- 9.9, respectively) compared with the control group (207.0 +/- 3.0) (p < 0.001). There were no significant differences in the plasmatic fibrinogen (PF) levels between the control and simvastatin treated groups (224.9 +/- 1.4 and 216.3 +/- 4.3, respectively). There were significant differences between the 30 or 60-day MI untreated groups compared with the 30 or 60-day multiple injuries + simvastatin treated group (p < 0.001). Endothelial denudation and intima widening were observed in the untreated injured groups, whereas in the 60 day multiple injuries group + simvastatin, a regression of histopathological lesions was observed. CONCLUSIONS: The decrease of the inflammatory component that would accompany early atherogenesis processes and the regression of the histopathological lesions after treatment could be attributed to the decreased plasmatic fibrinogen. PMID- 15887472 TI - Significance of serum troponin I elevation in patients with acute aortic dissection of the ascending aorta. AB - OBJECTIVE: This study evaluated the prevalence of increased cardiac troponin I (cTnI) in patients with acute aortic dissection of the ascending aorta (type A). METHODS AND RESULTS: In 119 consecutive patients with type A acute aortic dissection, serum cardiac troponin I was measured along with clinical, haemodynamic, electrocardiographic and echocardiographic variables obtained on admission. Cardiac troponin I was positive in 28 patients (23.5%; mean +/- SD: 6.1 +/- 14.7 ng/ml) and above the myocardial infarction threshold (1.5 ng/ml) in 12 (10%). Catecholamine infusion (17.9% vs. 4.4%; p = 0.03) and higher value of creatinine (35.7% vs. 15.4%; p = 0.03) were more frequent in patients with elevated troponin. Total mortality was 29.7% (n = 35) and surgical mortality was 16.8% (n = 17). An increased troponin was discriminatory with respect to mortality (OR: 4.1 (1.6-9.9); p = 0.002) in univariate analysis. However, this association was lost when other markers of death (age, stroke, ST-segment elevation, tamponade, catecholamine infusion, renal failure) were added in a multivariate model (OR: 2.2 (0.7-7.4); p = 0.19) indicating that the myocardial loss associated with troponin increase is not in itself a factor of mortality. CONCLUSIONS: Cardiac troponin I elevation is frequent in patients with type A aortic dissection. It might reflect a higher haemodynamic stress but does not necessarily reflect a negative prognosis. PMID- 15887473 TI - Percutaneous transcatheter closure of atrial septal defects: initial single centre experience and follow-up results. Initial experience with three dimensional echocardiography. AB - OBJECTIVE: Registry to report our single-centre experience in closing defects of the atrial septum. METHODS: Between 1996 and 2001, transcatheter device closure of significant atrial septal defects was performed in 32 adults (mean age 45.1 years, range from 15 to 76 years), using different device types. Six patients had a patent foramen ovale (PFO) and 26 patients had a secundum atrial septal defect (ASD II). Defects were selected by means of two-dimensional transoesophageal echocardiography (2D TOE). For 12 defects additional three-dimensional transoesophageal echocardiography (3D TOE) was performed. RESULTS: Mean balloon stretched diameter was 19.3 mm (range from 11 to 24 mm). A device was successfully placed in all PFO patients (100%) without complications at follow up. Successful immediate device placement occurred in 24 out of 26 ASD patients. Two immediate placement failures, 1 device embolisation and 1 device non fixation, occurred and required urgent surgery. There was 1 patient with pericardial effusion after the intervention, requiring pericardiocentesis. Three late placement failures occurred: 2 device embolisations and 1 device dislocation. Clinical success after 3 to 6 months follow-up, defined as the absence of shunt or small shunt, was achieved in all 6 PFO patients and in 21 out of 23 ASD patients (1 device embolisation, requiring surgery, occurred before follow-up was completed). Two patients with significant shunt, 1 late device dislocation and 1 late device embolisation, underwent elective surgery. For patient selection, 3D TOE proved useful in the evaluation of large and/or complex defects. CONCLUSION: Transcatheter device closure of carefully selected atrial septal defects is an alternative to surgery. Although the results are promising, surgical back-up is often needed. PMID- 15887474 TI - Urgent need to reorganize heart failure management: from paradoxes to heart failure clinics. AB - Despite the decreasing incidence of ischaemic heart disease and despite major medical advances in heart failure, the prevalence and mortality of chronic heart failure in the population is rising and the prognosis remains grim. Chronic heart failure is a complex disease, which is characterized by its progressive nature. In this paper, we approach the complexity of heart failure from four paradoxes: epidemiology, diagnosis, therapy and economical impact respectively. Taking these paradoxes into account, we formulate a number of essential components of alternative heart failure management programmes. Combating chronic heart failure requires the organization of centres for continuous care--as opposed to the traditional crisis intervention centres--preferably with a multidisciplinary structure to provide a "holistic approach" adapted to each patient's unique set of medical, psychosocial, physical and financial conditions. Patients taken care of in these novel multidisciplinary heart failure clinics have shown improved clinical status, decreased hospitalization rates, increased quality of life, longer life and lower costs. PMID- 15887475 TI - Immediate and six months clinical and angiographic results of intracoronary Ephesos stent implantation: the "Ephesos-1" study. AB - BACKGROUND: This study was conducted to evaluate the clinical and angiographic results of the Ephesos stent implantation in patients with symptomatic coronary artery disease. The Ephesos stent is a new balloon-expandable, stainless steel, tubular stent with multicellular design. METHODS: One hundred ten patients with native coronary artery disease were included in the study. The Ephesos stents were implanted in 163 de novo lesions detected in these patients. Immediate and long-term clinical and angiographic follow-up results were evaluated. RESULTS: Most of the patients had unstable angina 63.6%, and 36.7% of the lesions were type B and C. Mean lesion length was 12.7 +/- 4.7. In 62% of the patients the reference lumen diameter was < 3 mm. One Q-wave and one non-Q-wave myocardial infarction (MI) occurred due to acute thrombotic occlusion during hospital stay. The 6-month event-free survival rate was 77.3%. No patients died in the six-month follow-up period, but 2 patients had non-Q wave MI and 1 patient experienced Q wave MI within this period. Control angiographic data was collected from 110 patients (100% of patients and a total of 163 lesions). Angiographic restenosis rate was 18.1%. Twenty-two patients with restenosis had repeated target lesion balloon dilatation. CONCLUSION: The results of the present study showed that the Ephesos stent is a safe and effective choice with a low incidence of major adverse cardiac events and restenosis rate within six months of follow-up. PMID- 15887476 TI - Relation of insulin resistance and left ventricular function and structure in non diabetic patients with essential hypertension. AB - OBJECTIVE: Both left ventricular hypertrophy and insulin resistance (IR) have often been demonstrated in patients with essential hypertension (EH). Insulin may exert a direct growth-promoting effect on cardiomyocytes. The purpose of this study was to examine the relationship between left ventricular structure, function and IR in patients with EH. METHODS: We enrolled 73 patients (21 men, mean age 51.7 +/- 9.2 years) with untreated hypertension (BP > 140 and/or 90 mm Hg, fasting glycaemia < 110 mg/dl) and 64 healthy subjects without diabetes mellitus and hypertension (21 men, mean age 48.9 +/- 10.6 years) constituted the control group. In all subjects, transthoracic echocardiography was performed and blood samples were taken. Homeostasis model assessment (HOMA) was calculated by the formula: HOMA-index = fasting blood glucose (mg/dl) * immunoreactive insulin (microU/ml)/405 for the assessment of IR. Hypertensive patients were divided in two groups by mean HOMA index values. Each subject was examined for LV end diastolic diameter, septal and posterior wall thickness, LV mass index (LVMI), fractional shortening (FS), mitral inflow velocity pattern, atrial filling fraction (AFF), left ventricular outflow velocity pattern and the total ejection isovolume index (TEI index). RESULTS: The HOMA index (p < 0.001), LVMI (p < 0.001), AFF (p < 0.0001), peak A velocity (p < 0.028), septal (p < 0.0001) and posterior (p < 0.0001) wall thickness were significantly higher and FS (p < 0.001), E/A ratio (p < 0.0001) were significantly lower in hypertensive patients than healthy controls. LVMI (p < 0.01) and septal wall thickness (p < 0.001) were significantly higher in those hypertensive patients with a higher HOMA index. The HOMA-index was univariately related to the TEI index (r = 0.27, p = 0.01) and septal wall thickness (IVS) (r = 0.29, p = 0.01) by Pearson correlation analysis in hypertensive patients. LVMI, FS and mitral inflow velocity pattern were not related to the HOMA index. The TEI index (R2 = 0.20, p = 0.0001) and IVS (R2 = 0.12, p = 0.002) were significantly related to the HOMA-index as an independent variable by stepwise regression analysis. CONCLUSIONS: These results demonstrated that hypertensive patients had both abnormal cardiac structure and function and higher IR index. In our study group, the effect of hypertension on cardiac structure and function was correlated with IR. Our results suggested that IR might be an important factor causing left ventricular dysfunction and wall thickness in non-diabetic patients with EH. PMID- 15887477 TI - Randomised evaluation of a specific training of general practitioners in cardiovascular prevention. AB - OBJECTIVE: A lack of global cardiovascular (CV) risk approach is often observed in the clinical practice. Having conceived a clinical strategy aimed at helping general practitioners (GPs) to assess and manage global CV risk in the daily practice, we wanted to evaluate the impact of a training promoting this strategy. METHODS: Randomised controlled trial involving GPs registered in a continual medical education (CME) group. The intervention and control arms included 7 (123 GPs) and 13 (220 GPs) groups, respectively. The intervention was a training in CV prevention provided during a CME meeting. GPs answered a questionnaire four months after the training. The main outcome was the use of a global CV risk assessment tool. Secondary outcomes were the GPs' awareness of the CV diseases burden, knowledge of the CV risk factors, and CV therapeutic attitudes. RESULTS: More trained GPs (76% vs. 52%) used a global CV risk assessment tool (p = 0.003). Significant differences were also observed for secondary outcomes: (a) awareness of CV diseases burden: more trained GPs were aware of the lethality from myocardial infarction (37% vs. 21%, p = 0.047) and considered it as the main cause of sudden death in adults (78% vs. 59%, p = 0.018); (b) knowledge of risk factors: more trained GPs considered familial CV history (89% vs. 73%, p = 0.021) and HDL-cholesterol (82% vs. 62%, p = 0.013) as important CV risk factors; (c) therapeutic attitudes: fewer trained GPs used to prescribe a lipid-lowering drug based on a total cholesterol value without considering the other risk factors (44% vs. 65%, p = 0.013). CONCLUSIONS: The global CV risk approach in the general practice was favourably influenced by this training provided during a CME meeting. The impact of the training on patients' outcomes should be addressed in a prospective trial. PMID- 15887478 TI - Long-term safety of intracoronary haemodynamic assessment for deferral of angioplasty in intermediate coronary stenoses: a 5-year follow-up. AB - BACKGROUND: Intracoronary flow and pressure measurements can be used for evaluating intermediate lesions. Studies focussing on short- and medium-term results demonstrated its safety. Long-term results are, however, not available. OBJECTIVE: The goal of this study was to assess the long-term safety and clinical implications of decision making for intermediate coronary stenosis based on intra coronary haemodynamic measurements. METHODS AND RESULTS: In this prospective study, 61 patients with an intermediate coronary stenosis were included between January 1994 and December 1998. In these patients either coronary flow reserve or fractional flow reserve was measured. Death, target vessel revascularization, myocardial infarction, unstable angina and cerebrovascular accident were considered as major adverse cardiac events. The patients were followed during 5.5 (1.8) years for the occurrence of major adverse cardiac events (MACE). Although many patients presented with complaints, only 19.7% experienced a MACE in the follow-up period. CONCLUSION: Intracoronary measurements of CFR and FFR can be routinely used for objective clinical decision making in intermediate coronary stenoses. The low 5-year event rate supports conservative treatment strategy when cut-off values are implemented. PMID- 15887479 TI - Postinfarction ventricular septal rupture: surgical intervention and risk factors influencing hospital mortality. AB - Postinfarction rupture of the interventricular septum is usually fatal without surgical intervention and requires urgent closure. Between 1989 and 2003 twenty consecutive patients (15 male, 5 female), underwent postinfarction ventricular septal rupture (VSR) repair. Mean age of the patients was 62.05 +/- 7.51 years. Fifteen patients were operated within 48 hours after myocardial infarction. Patch reconstruction was performed in all patients. Infarct locations were anterior in 65%, posterior in 35%. Coronary artery surgery was performed in 14 patients (70%). Hospital mortality was 30% (6 patients). Four patients were presented for surgical therapy with frank cardiogenic shock or low cardiac output syndrome. A residual shunt was detected in 4 patients and three of these patients were reoperated. One of them, who has been reoperated on the first day of the postoperative period, did not survive. The statistical analysis of the patients' records demonstrated that time period between MI and surgery, applied additional CABG procedure, the sex of the patients and the site of the rupture are significant factors influencing in-hospital mortality. Preoperative condition, age of the patients and the number of the affected coronary vessels do not have an important effect on the mortality. Postinfarction ventricular septal rupture is a fatal complication of the myocardial infarction and must be treated surgically. The time interval between septal rupture independent from the preoperative haemodynamic condition, the location of the defect and additional myocardial revascularization procedure are the factors influencing the early outcome. PMID- 15887480 TI - Lacrimo-auriculo-dento-digital syndrome with QT prolongation. PMID- 15887481 TI - Acute migraine attack, angina-like chest pain with documented ST-segment elevation and slow coronary flow. AB - Slow flow of dye in epicardial coronary arteries is not an infrequent finding in patients during routine coronary angiography. The coronary slow flow phenomenon is an angiographic finding characterized by delayed distal vessel opacification in the absence of significant epicardial coronary artery disease. It is speculated that coronary slow flow is a new disease characterized by acute but recurrent perturbations of microvascular function. There are many theories concerning the pathogenesis of migraine. The clinical effectiveness of vasoactive drugs and many investigations on the cerebral blood flow in patients with migraine, strongly support a vascular theory. The relationship between migraine and cardiopathy has not been sufficiently established and controversy exists concerning its favouring role in coronary artery disease. We report a case of an acute migraine attack in a patient who uses triptans (5-HT(1B/1D) receptor agonists). The attack was accompanied by angina-like chest pain with documented ST-segment elevation and slow coronary flow in the absence of any significant obstructive coronary artery disease and no evidence of any major epicardial coronary arterial spasm. PMID- 15887482 TI - Successful percutaneous coronary intervention for acute myocardial infarction caused by simultaneous occlusion of two major coronary arteries in patients with diabetes mellitus. A report of two cases. AB - Percutaneous coronary intervention (PCI) for acute myocardial infarction (AMI) is a common therapeutic method. Although two or more culprit lesions are rarely observed simultaneously in AMI patients, we present two cases of AMI caused by simultaneous occlusion of two major coronary arteries, the left anterior descending and right coronary arteries. In both cases, emergency PCI for the two major vessels was successful. Both patients had type 2 diabetes mellitus, which might have contributed to simultaneous occlusion of the two coronary arteries. PMID- 15887483 TI - Right atrial thrombosis in a heart transplant recipient after initiation of sirolimus. PMID- 15887484 TI - Fluorescence probe assisted post-column detection for lipid analysis in microbore LC. AB - A general approach, still few exploited so far and never associated with microbore-LC, consisting of detection of various lipid classes (i.e. phospholipids, triglycerides, ceramides and glycosphingolipids) by non-covalent association with 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence probe is developed. This mode of detection was coupled with non-aqueous reversed-phase microbore-LC (C18) by using classical post-column fluorescence detection. The classical LC system was first adapted to microbore-chromatography (internal diameter 1 mm) without apparatus miniaturization of the solvent delivery system and the detection cell. For this purpose, the detection parameters (probe concentration, post-column flow rate, post-column reactor length and post-column system temperature) were optimized by a central composite design (CCD) using a mixture of phosphatidylcholine (PC) species as a lipid model and DPH (lambda(ex) = 350 nm, lambda(em) = 430 nm) as a fluorescence probe. The optimal conditions of detection for the various molecular species of PC were determined for a DPH concentration of 3.35 micromol/L, a post-column flow rate of 0.5 mL/min, a reactor length of 1.4 m and a temperature of 35 degrees C. The fluorescence response was linear over a wide range of PC species from 5 microg/mL to 100 microg/mL and the lower limit of detection (signal/noise = 3) was about 1 microg/mL, that is equivalent to evaporative light scattering detection (ELSD). Others molecular species of various classes of lipids, i.e. triglycerides, ceramides and glycosphingolipids were also easily detected. Thus, this study demonstrated the versatility of the proposed system of detection which was shown to be sensitive, easy to perform, non-destructive and allowed, in contrast to ELSD, for a linear response with various polarity lipid classes. PMID- 15887485 TI - Modeling Caco-2 permeability of drugs using immobilized artificial membrane chromatography and physicochemical descriptors. AB - This study evaluates the potential of immobilized artificial membrane (IAM) chromatography, in combination with other physicochemical descriptors for high throughput absorption profiling during lead optimization. An IAM chromatographic method was developed and validated. Absorption profiles of 32 structurally diverse compounds (acidic, basic, neutral and amphoteric) were then evaluated based on their IAM retention factor (log k'IAM), molecular weight (MW), calculated log P (C log P), polar surface area (PSA), hydrogen bonding capacity (HBD and HBA) and calculated Caco-2 permeability (QPCaco). Using regression and stepwise regression analysis, experimental Caco-2 permeability was correlated against log k'IAM and a combination of various physicochemical variables for quantitative structural-permeability relationship (QSPR) study. For the 32 structurally diverse compounds, log k'IAM correlated poorly with Caco-2 permeability values (R2 = 0.227). Stepwise regression analysis confirmed that Clog, PSA, HBD and HBA parameters are not statistically significant and can be eliminated. Correlation between Caco-2 cell uptake and log k'IAM was enhanced when molecular size factor (MW) was included (R2 = 0.555). The exclusion of 11 compounds (paracellularly and actively transported, Pgp substrates and blocker, and molecules with MW lesser than 200 and greater than 800) improved the correlation between Caco-2 permeability, IAM and MW factors to R2 value of 0.84. The results showed that IAM chromatography can only profile the passive absorption of drug molecules. Finally, it was confirmed in this study that the IAM model can accurately identify the Caco-2 permeability of nontransported Pgp substrates, such as verapamil and ketoconazole, through passive permeation because of their high permeability. IAM chromatography, combined with molecular size factor (MW), is useful for elucidating biopartitioning mechanism of drugs. PMID- 15887486 TI - Role of ion pairing in anionic additive effects on the separation of cationic drugs in reversed-phase liquid chromatography. AB - Mobile phase additives can significantly affect the separation of cationic drugs in reversed-phase liquid chromatography (RPLC). Although there are many applications for anionic additives in RPLC separations, the retention mechanism of basic drugs in the presence of inorganic and highly hydrophilic anionic species in the mobile phase is not at all well understood. Two major retention mechanisms by which anionic additives can influence the retention of cations are: (1) ion pair formation in the mobile phase with subsequent retention of the neutral ion pair; (2) pre-sorption of anionic additives on the stationary phase followed by "dynamic ion-exchange" or "electrostatic interaction" with the analytes. Because the use of ion pair chromatography in the separation of proteins, peptides, and basic drugs is rapidly increasing, understanding the retention mechanism involved is becoming more important, especially for the smaller commonly used hydrophilic anionic additives (e.g., formate HCOO, chloride Cl-, trifluoroacetate CF3COO-, perchlorate ClO4-, and hexafluorophosphate PF6-). In this work, we compared various anionic additives in light of their effects on the retention of basic drugs. As did many others we found that the addition of anionic additives (Cl-, CF3COO-, ClO4-, PF6-) profoundly influences the retention of basic drugs. In order to explain the data and differentiate the mechanisms by which the anionic additives perturb the chromatography, we used ion pair formation constants independently measured by capillary electrophoresis (CE) under the mobile phase conditions (pH, solvent composition) identical to those used in chromatography. Agreement between the predicted and experimental chromatographic data under various conditions was evaluated. Under specific circumstances (e.g., pH, stationary phase, and nature of anionic additive), we conclude that the ion pair mechanism is more important than the dynamic ion exchange and at other conditions it remains a significant contribution. PMID- 15887487 TI - Simultaneous quantification of differently glycosylated, acetylated, and 2,3 dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one-conjugated soyasaponins using reversed-phase high-performance liquid chromatography with evaporative light scattering detection. AB - A novel method utilizing high-performance liquid chromatography (HPLC) with evaporative light scattering detection (ELSD) and electrospray ionisation mass spectrometry (ESI-MS) was developed for the analysis of soyasaponins, a divers group of triterpenic compounds with one or two sugar side chains, occurring in soy. Group A soyasaponins in different degrees of acetylation, as well as group B soyasaponins in both their 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)-conjugated and non-conjugated forms could be separated and quantified using authentic soyasaponin standards, in one single run. The method was tested by the determination of the soyasaponin content and composition of eight soygerm samples of different origin. Differences in the composition and the degree of acetylation of the group A soyasaponins were observed among these samples. The group B soyasaponins showed much less variability and they were mainly present in their DDMP-conjugated form. PMID- 15887488 TI - Isolation and purification of coumarin compounds from Cortex fraxinus by high speed counter-current chromatography. AB - High-speed counter-current chromatography (HSCCC) was successfully used for the isolation and purification of coumarin compounds from Cortex fraxinus, the Chinese herbal drug. n-Butanol-methanol-0.5% acetic acid (5:1.5:5, v/v) was used as the two-phase solvent system. 14.3 mg of fraxin, 26.5 mg of aesculin, 5.8 mg of fraxetin and 32.4 mg of aesculetin with the purity of 97.6, 99.5, 97.2 and 98.7%, respectively were obtained from 150 mg of crude extracts of C. fraxinus in a single run. The structures of the isolated compounds were identified by 1H NMR and 13C NMR. PMID- 15887489 TI - Use of lipophilic ion adsorption isotherms to determine the surface area and the monolayer capacity of a chromatographic packing, as well as the thermodynamic equilibrium constant for its adsorption. AB - A method that champions the approaches of two independent research groups, to quantitate the chromatographic stationary phase surface available for lipophilic ion adsorption, is presented. For the first time the non-approximated expression of the electrostatically modified Langmuir adsorption isotherm was used. The non approximated Gouy-Chapman (G-C) theory equation was used to give the rigorous surface potential. The method helps model makers, interested in ionic interactions, determine whether the potential modified Langmuir isotherm can be linearized, and, accordingly, whether simplified retention equations can be properly used. The theory cultivated here allows the estimates not only of the chromatographically accessible surface area, but also of the thermodynamic equilibrium constant for the adsorption of the amphiphile, the standard free energy of its adsorption, and the monolayer capacity of the packing. In addition, it establishes the limit between a theoretical and an empirical use of the Freundlich isotherm to determine the surface area. Estimates of the parameters characterising the chromatographic system are reliable from the physical point of view, and this greatly validates the present comprehensive approach. PMID- 15887490 TI - Use of temperature programming to improve resolution of inorganic anions, haloacetic acids and oxyhalides in drinking water by suppressed ion chromatography. AB - Temperature programming was used to improve selectivity in the suppressed ion chromatographic (IC) separation of inorganic anions, haloacetic acids and oxyhalides in drinking water samples when using NaOH gradient elution. The programme exploited varying responses of these anions to changes in temperature. Heats of adsorption (deltaH, kJ/mol) for 17 anionic species were calculated from van't Hoff plots. For haloacetic acids, both the degree of substitution and log P (log of n-octanol-waterpartition coefficient) values correlated well with the magnitude of the temperature effect, with monochloro- and monobromoacetic acids showing the largest effect (deltaH= -10.4 to -10.7 kJ/mol), dichloro- and dibromoacetic acids showing a reduced effect (deltaH= -6.8 to -8.4 kJ/mol) and trichloro-, bromodichloro- and chlorodibromoacetic acids showing the least effect (deltaH= -4.7 to -2.4 kJ/mol). The effect of temperature on oxyhalides ranged from deltaH= 8.4 kJ/mol for perchlorate to deltaH= -9.1 kJ/mol for iodate. The effectiveness of two commercial column ovens was investigated for the application of temperature gradients during chromatographic runs, with the best system applied to improve the resolution of closely retained species at the start, middle and end of the separation obtained using a previously optimised hydroxide gradient, in a real drinking water sample matrix. Retention time reproducibility of the final method ranged from 0.62 to 3.18% RSD (n = 30) showing temperature programming is indeed a practically important parameter to manipulate resolution. PMID- 15887491 TI - Large-scale isolation of flavan-3-ol phloroglucinol adducts by high-speed counter current chromatography. AB - Flavan-3-ol phloroglucinol adducts were synthesised through acid catalysed degradation of a procyanidins-rich grape seed extract in the presence of phloroglucinol. The reaction mixture (3.3 g) was fractionated without further sample preparation using the all-liquid chromatographic technique of high-speed counter-current chromatography (HSCCC). Selected solvent systems were hexane ethyl acetate-methanol-water (0.1:5:0.1:5, v/v/v/v) and (1.5:10:1.5:10, v/v/v/v). The fractions obtained were found to contain almost pure compounds, in some cases final purification was achieved by preparative HPLC. The so-obtained pure standards of (+)catechin-(4alpha-->2)-phloroglucinol, (-)epicatechin-(4beta-->2) phloroglucinol, (+)catechine, (-)epicatechin-3-O-galloyl-(4beta-->2) phloroglucinol, (-)epicatechin, and (-)epicatechin gallate are required for quantification of acid-catalysed phloroglucinol degradation products of procyanidins. PMID- 15887492 TI - Large-bore particle-entrapped monolithic precolumns prepared by a sol-gel method for on-line peptides trapping and preconcentration in multidimensional liquid chromatography system for proteome analysis. AB - The present report describes the preparation and characterization of large-bore particle-entrapped monolithic precolumns, which are suitable for incorporation into a two-dimensional liquid chromatography (2D-LC) system for proteome analysis. The fritless precolumns with different inner diameter (i.d.) (320 and 530 microm) were rapidly and successfully prepared by entrapping octadecylsilica (ODS) particles (5 microm, 300 A) prepacked into fused silica capillaries with a sol-gel network, which was formed by hydrolysis and polycondensation of methyltriethoxysilane (MTES). By optimizing the composition of the sol solution, the resulting large-bore monolithic precolumns of 5 mm length allow a flow rate of 20 microL/min loading buffer at a reasonable low back pressure of 25 bar or less and are capable of withstanding up to 300 bar inlet pressure. Scanning electron micrograms of the precolumns profile showed that the evolving sol-gel network joined particles to each other and onto the column wall, and no cracking or shrinkage of the column bed was observed even in 530 microm-i.d. capillary. The performance of the particle-entrapped monolithic precolumns used for preconcentration and desalting of proteolytic digest was evaluated by on-line coupling the large-bore precolumns with a capillary reversed-phase liquid chromatographic (RPLC) column followed by UV detection. The laboratory-made monolithic precolumns with 320 and 530 microm i.d. were characterized by using BSA tryptic digest or peptide standards as the analytes with respect to sample loading capacity, linearity, recovery and reproducibility, etc. The results indicate that the large-bore and short precolumns (5 mm x 320 microm i.d. or 5 mm x 530 microm i.d.) allow sample fast loading at a flow rate of 30 or 60 microL/min. The precolumns also have a mass loading capacity for BSA peptides of about 70 microg and for standard peptides of about 80 microg. Good linear calibration curves (R2 > 0.99) were obtained and the limits of detection (signal to-noise ratio, S/N = 3) were improved by more than 60-fold and were between 0.53 and 1.32 ng/microL even with a UV absorbance detector. The total recovery was found to be approximately 90-100% for BSA digest and standard peptides. The day to-day relative standard deviation (RSD) values for recoveries of BSA peptides on a single precolumn ranged from 4.66 to 7.56% and 2.68 to 3.05% for precolumn back pressure, while the column-to-column RSD values were 3.51-6.13% and 1.22-1.26% for recoveries of BSA peptides and precolumn back pressure, respectively. With good precolumn reproducibility, no significant degradation or decrease in precolumn performance was showed even after approximately 150 preconcentration/desorption cycles. The precolumns also proved to be resistant to salt buffer with high concentration and low-pH mobile phase. The large-bore particle-entrapped monolithic precolumns will be further used in a high throughput 2D-LC array system coupled with tandem matrix assisted laser desorption/ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF TOF-MS) detection for proteome analysis. PMID- 15887493 TI - Development and validation of methods for the trace determination of phthalates in sludge and vegetables. AB - A routine method which is simple, quick and precise has been set up and validated for phthalate analysis in environmental samples (tomato plants and sewage sludges). Six phthalates have been studied simultaneously: dimethylphthalate, diethylphthalate, di-n-butylphthalate, n-butylbenzylphthalate, di-2-ethyl-hexyl phthalate (DEHP) and di-n-octylphthalate. Optimization of sample, solvent extraction uses a Soxtec apparatus and extract purification with an a solid-phase extraction cartridge allows between 90 and 110% recovery of phthalates. Precise, sensitive and selective identification and quantifying of analytes is by GC-MS in the single ion monitoring mode. This protocol allows analytes with concentrations as low as 10 microg/kg dry matter (DM) to be determined from small (1-2 g DM) samples. This analytical method has been applied to the phthalate transfer study for agricultural recycling of sludges, where phthalate bioavailability has been studied in aquiculture using two types of experiments. Tomatoes have been grown in containers where the trace organics have been directly introduced as pure substances, and in a second experiment under the same growth conditions, sewage sludge has replaced the pure substances. Transfer of these trace organics has been followed into the various parts of the tomato plant and in general only the DEHP is worthy of note although its percentage transfer remains very low even in an experiment designed to maximize this. PMID- 15887494 TI - Microtrap modulated flame ionization detector for on-line monitoring of methane. AB - A microtrap for on-line analysis of trace methane was developed. Silicosteel tubing, 15 cm long, 1.02 mm i.d. and 1.59 mm o.d. was packed with Carbosphere 80/100 mesh and placed between the standard gas source and the flame ionization detector (FID) to be used as a preconcentrator and an injector. The sample stream was passed continuously through the microtrap that was heated at a fixed interval by an electric pulse. To achieve the highest adsorption and desorption efficiency, parameters affecting the response of the system were optimized, and adsorption temperature in the range of 25 to -50 degrees C was studied. The response from the microtrap as compared to that from direct-flame ionization detector was enhanced by as much as 260 times. The on-line microtrap system showed good linearity (r2 > 0.99), low detection limit (28.3 ppbv) and good long term stability (relative standard deviation, RSD of less than 5.0%). Therefore, this simple device is suitable for on-line analysis of trace methane and similar small molecules in the environment as in high purity gases. PMID- 15887495 TI - Enantiomeric determination, validation and robustness studies of racemic citalopram in pharmaceutical formulations by capillary electrophoresis. AB - A chiral capillary electrophoresis (CE) method has been developed allowing the enantiomeric separation of racemic citalopram (R-(-) and S-(+) citalopram) using as chiral selector carboxymethyl-gamma-cyclodrextrin (CM-gamma-CD). The influence of chemical and instrumental parameters on the separation such as cyclodextrin (CD) and buffer concentrations, buffer pH, voltage, injection pressure, ..., was investigated. Good chiral separation of the racemic mixture was achieved in less than 4 min using a fused-silica capillary and as background electrolyte (BGE) a phosphate buffer solution (20 mM, pH 7) containing 0.15% (w/v) of CM-gamma-CD as chiral selector. The separation was driven in normal polarity mode at 15 degrees C, 30 kV and hydrodynamic injection. In order to validate the method, the stability of the solutions, precision (repeatability, reproducibility and F Snedecor test), linearity (Lack of Fit and ANOVA tests) accuracy (98-101%), detection and quantitation limits (0.06 and 0.2 mg L(-1), respectively), on a selected analytical placebo, were examined. Besides, a robustness test was performed using the Plackett-Burman fractional factorial experimental design using a matrix of 15 experiments for seven factors (internal parameters) with a statistical treatment suggested by Youden and Steinner. The proposed method is fast, sensitive, inexpensive and, besides, it has been evaluated by means of an extensive validation study and an exhaustive robustness test. The scope of this validated and robust method has been proved in the analysis of four pharmaceutical formulations; two of them (recently available in Spain), which just contained S-(+)-citalopram (escitalopram) as active principle. Recoveries between 101 and 103%, with regard to their nominal contents were obtained. In the other two pharmaceutical ones, the method provided the separation and quantification of both chiral isomers in the existing racemic mixture. PMID- 15887496 TI - Method for on-line derivatization and separation of aspartic acid enantiomer in pharmaceuticals application by the coupling of flow injection with micellar electrokinetic chromatography. AB - A novel, easy and accurate capillary electrophoresis (CE) coupled with flow injection (FI) method for the separation and determination of aspartic acid (Asp) enantiomers by on-line derivatization had been developed, and it had been applied to the real sample for the first time. The derivatization reagents were o phthalaldehyde (OPA) and mercaptoethanol (ME), which were obtained easily, the chiral selector was beta-cyclodextrin (beta-CD), the micellar chemical was sodium dodecyl sulfate (SDS), and the modifier was methanol. By on-line derivatization, aspartic acid enantiomers were automatically and reproducibly converted to the ultraviolet (UV)-absorbing diastereoisomer derivates, which were separated by micellar electrokinetic chromatography (MEKC). According to the factors affecting the separation and sensitivity of aspartic acid enantiomer and other amino acids in the real sample, the pH value and concentration of the buffer, the concentration of beta-CD and SDS, the volume percentage of the methanol (v/v) in the buffer, the applied voltage and the conversion time were selected as the investigating variates. Under the investigated separation conditions, D-aspartic acid (D-Asp), L-aspartic acid (L-Asp) and other four amino acids achieved the baseline separation in not only the standard mixture of amino acids but also the real sample (Compound Amino Acid Injection (6AA)). The repeatability (defined as relative standard deviation (RSD), n = 5) was 4.0% and 4.0% with peak area evaluation, and 4.2% and 3.7% with peak height evaluation for D-Asp and L-Asp in the real sample. Recovery at added standard levels of 1.0, 3.0 and 6.0 mM was 92%, 104% and 109%, respectively. PMID- 15887497 TI - Determination of rate constants and activation energy of 3-chloro-1,2-propanediol hydrolysis by capillary electrophoresis with electrochemical detection. AB - A method based on capillary electrophoresis with electrochemical detection (CE ED) to calculate the rate constants and activation energy of 3-chloro-1,2 propanediol (3-MCPD) hydrolysis was described. Effects of several factors, such as the pH value and the concentration of the running buffer, separation voltage, injection time and the potential applied to the working electrode, were investigated to find the optimum conditions. With a 50 cm length of 25 microm diameter fused-silica capillary at a separation of 10 kV, well-defined separation of 3-chloro-1,2-propanediol from glycerol was achieved in 30 mmol/l borax (pH 9.24) within 13 min. Operated in a wall-jet configuration, a 328 microm copper disk electrode used as the working electrode exhibits good response at 0.65 V (versus SCE) for 3-MCPD and glycerol. The rate constants of 3-MCPD hydrolysis at different temperatures were determined by monitoring the concentration changes of 3-MCPD. At 80, 85 and 90 degrees C, the measured rate constants of 3-MCPD hydrolysis were 3.8 x 10(-3) min(-1), 7.1 x 10(-3) min(-1) and 11.5 x 10(-3) min( 1), respectively. The activation energy for 3-MCPD hydrolysis was calculated to be 118.1 kJ/mol, which is in good agreement with the value in the literature. PMID- 15887498 TI - Indirect fluorescence detection of amino sugars with the use of copper complexes of tryptophan and its analogues following high-performance liquid chromatographic separation. AB - A simple, indirect fluorescence detection method has been developed for detecting specific mono-amino sugars (D-glucosamine, D-galactosamine, D-mannosamine) following chromatographic separation. The eluting amino sugars release L tryptophan (L-Trp) from a copper-tryptophan complex which is introduced postcolumn. Analyte detection is based on measuring the increase in L-Trp fluorescence, which is quenched when complexed with copper. Two tryptophan analogues, 5-hydroxy-L-tryptophan (5-HTP) and DL-5-methoxytryptophan (5-MTP), were also evaluated as postcolumn reagents. 5-MTP was found to be a suitable alternative to L-Trp for the detection of these mono-amino sugars. Detection limits for D-glucosamine, D-galactosamine, and D-mannosamine are in the range of 0.15-0.30 nmol injected. PMID- 15887499 TI - Enantioseparation of 2-O-beta-D-glucopyranosyl-2H-1,4-benzoxazin-3(4H)-one and its 7-chloro-derivative by capillary zone electrophoresis using native and substituted beta-cyclodextrins as chiral additives. AB - Efficient, rapid and inexpensive methods were established for the chiral separation of two glucopyranosyl compounds from plant extracts, by capillary zone electrophoresis (CZE). Baseline separation was achieved for both compounds. Several native cyclodextrins and their derivatives were tried as chiral selectors. CM-beta-CD and HP-beta-CD (with addition of acetonitrile in the buffer) gave rise to optimal chiral separation for the two compounds, respectively, each within a few minutes. The effects of several parameters on the chiral separation were studied. PMID- 15887500 TI - The dentist of tomorrow. PMID- 15887501 TI - Effect of residual caries-disclosing solutions on microleakage of a dental adhesive system. AB - OBJECTIVE: The purpose of this in vitro study was to determine the effect of residual caries-disclosing dyes on a dentinal adhesive system and accompanying resin composite, using microleakage as the quantifying criterion. METHOD AND MATERIALS: The dye pretreatment groups (10 teeth in each group, n = 60) were as follows: Group 1, Caries Detector; group 2, Cari-d-tect; group 3, Seek; group 4, Snoop; group 5, To Dye For; and group 6, control (no dye pretreatment). Circular preparations were cut on the facial or lingual surface of each tooth. Caries disclosing dyes were placed on the preparations; this was followed by treatment with 35% phosphoric acid etchant, application of a Single-Bond adhesive, and insertion of Z-100 hybrid resin composite. Following thermocycling, the teeth were sealed with nail varnish, placed in methylene blue dye, invested in acrylic resin, labeled, and sectioned. Leakage was examined (20x magnification) by dye penetration at the occlusal and gingival surface positions of the specimen blocks using an ordinal ranking system. The data were analyzed using two-way analysis of variance with P < .05 as the significance level. RESULTS: Results of the study showed that, at the occlusal surface position of the restorations, significantly greater leakage was exhibited by To Dye For compared to Snoop. At the gingival surface position, significantly greater leakage was exhibited by: (1) Caries Detector, Cari-d-tect, Seek, and To Dye For compared to the controls; and (2) To Dye For compared to Snoop. When comparing the same dye pretreatment groups with respect to occlusal versus gingival surface positions, significantly greater leakage was exhibited at the gingival surface positions, except for the control group. CONCLUSION: Our data showed that all dye groups leaked (except Snoop at the occlusal surface position), and significantly greater leakage was exhibited at the gingival surface position of all groups except the control. Snoop revealed the least leakage of all the dyes tested. PMID- 15887502 TI - A microleakage study of single-bottle adhesives applied to enamel and cementum and aged by both occlusal loading and thermocycling. AB - OBJECTIVE: To compare enamel and dentin microleakage of three single-bottle dentin-bonding agents (DBA) (One-Step, Single Bond, and Dentamed P&B) on teeth that were then subjected to thermocycling and occlusal loading. METHOD AND MATERIALS: Class 5 cavities were prepared on the buccal and lingual surfaces of 30 extracted human molars and divided into three groups. The occlusal margins were in enamel, and the cervical margins were in cementum. DBA were applied strictly according to the manufacturers' recommendations, and the preparations were restored with resin composite. Specimens were subjected to 2,000 intermittent occlusal loads and 3,000 thermocycles and then immersed in 0.5% basic fuchsin dye for 72 hours. Four consecutive sections were cut for each specimen and examined under a stereomicroscope. The extent of dye penetration was measured and recorded using a nonparametric scale from 0 to 4. RESULTS: The Mann Whitney U test showed a significant difference at cementum margins between One Step and Dentamed P&B, as well as between Single Bond and Dentamed P&B, and an insignificant difference between Single Bond and One-Step. There was no significant difference in dye penetration at enamel margins between the three DBA. CONCLUSION: Under combined use of occlusal loads and thermocycling, the sealing ability of single-bottle DBA at dentin margins ranged from good for One Step to moderate for Single Bond to poor for Dentamed P&B. At enamel margins, all materials performed equally well. PMID- 15887503 TI - Use of laser fluorescence in monitoring the durability and cariostatic effects of fluoride and chlorhexidine varnishes on occlusal caries: a clinical study. AB - OBJECTIVE: The aim of this study was to compare the effectiveness and caries preventive effects of a fluoride varnish (Fluor Protector; 0.1% fluoride) and a chlorhexidine varnish (Cervitec; 1% chlorhexidine and 1% thymol) on initial caries of occlusal fissures and to monitor the durability of the varnishes by the use of a fluorescence system, DIAGNOdent (Dd). METHOD AND MATERIALS: Nineteen subjects underwent professional tooth cleaning at the beginning of the study. Afterward, fluorescence measurements (baseline) were obtained from 56 test sites and 28 control sites on the central fossae of molars. Each subject had Fluor Protector and Cervitec application on randomly assigned experimental teeth, and applications were followed by Dd measurements. Measurements were repeated after 1 and 6 months. RESULTS: There was an increase in fluorescence values immediately after application of varnish in the two test groups. One month after varnish application, Dd scores for group Fluor Protector were not significantly different from Dd scores obtained immediately after application (P = .108), whereas Dd scores for group Cervitec decreased significantly (P = .007). For both test groups, there was a significant decrease between the values obtained immediately after application and those acquired after 6 months. There was also a significant decrease between 1 month and 6 months for test groups and a slight but significant increase for the control group (P = .011). No significant difference was found between the Fluor Protector and Cervitec groups after 6 months. CONCLUSION: It was concluded that the Dd system may be used to monitor the existence of these two varnishes. PMID- 15887504 TI - Porcelain veneers as an alternative for the esthetic treatment of stained anterior teeth: clinical report. AB - The esthetic treatment of anterior teeth with porcelain veneers is a conservative and lasting treatment option. When the tooth is severely discolored, the clinician must choose a ceramic system very carefully, because translucent materials, such as hot-pressed ceramic, require excessive reduction of the dental structure to mask the discoloration. This article describes a technique in which discolored teeth were masked in a conservative and esthetically pleasing way with feldspathic porcelain veneers. PMID- 15887505 TI - Onset and duration periods of articaine and lidocaine on maxillary infiltration. AB - OBJECTIVES: The aim of this study was to compare the onset and duration of pulpal anesthesia by maxillary infiltration using 2% lidocaine with 1:100,000 epinephrine, 4% articaine with 1:200,000 epinephrine, and 4% articaine with 1:100,000 epinephrine. METHOD AND MATERIALS: Twenty healthy patients randomly received 1.8 mL of one of the three local anesthetics during operative dentistry procedures of low complexity on three maxillary posterior teeth. Onset and duration were determined using an electric pulp tester. RESULTS: The mean values for pulpal onset were 2.8, 1.6, and 1.4 minutes and for pulpal duration were 39.2, 56.7, and 66.3 minutes, respectively, for 2% lidocaine with 1:100,000 epinephrine, 4% articaine with 1:200,000 epinephrine, and 4% articaine with 1:100,000 epinephrine. Statistical analysis by the Kruskal-Wallis nonparametric test showed significant differences with better results (shorter onset and longer duration periods) for both articaine solutions compared with the lidocaine solution. Although 4% articaine with 1:100,000 epinephrine clinically presented the shortest onset and the longest duration periods, there was no statistically significant difference between the articaine solutions. CONCLUSION: Both articaine solutions produced shorter onset and longer duration of pulpal anesthesia by maxillary infiltration than the lidocaine solution did. Statistical analysis did not confirm better clinical results of 4% articaine with 1:100,000 epinephrine than with 4% articaine with 1:200,000 epinephrine. PMID- 15887506 TI - Distribution of different morphologic types of subgingival calculus on proximal root surfaces. AB - OBJECTIVES: To evaluate the distribution of morphologic types of subgingival calculus at different parts of the proximal root surface. METHOD AND MATERIALS: Ninety extracted teeth from 29 chronic periodontitis patients were collected. The mean pocket depths and periodontal attachment levels of the extracted teeth were 5.93 +/- 1.51 mm and 7.82 +/- 1.75 mm, respectively. The proximal root surfaces below the gingival margin were divided into three parts in an apicocoronal direction, and each of these portions was further divided into three parts in a buccolingual direction. Subgingival calculus present was classified as: 1 = crusty, spiny, or nodular; 2 = ledge or ring; 3 = thin, smooth veneers; 4 = finger- or fernlike; 5 = individual calculus islands/spots; or 6 = supramarginal upon submarginal deposits. The distribution of different morphologic types of subgingival calculus on each division of the mesial and distal proximal root surfaces was evaluated with a magnifier. RESULTS: Regardless of the morphologic type, calculus deposits were observed at around 30% of proximal root surfaces. The coronal thirds of the root surfaces were found to have significantly more calculus deposits than the middle thirds (P < .05). In general, it was observed that most of the deposits were of the thin, smooth veneer type on all root surfaces. CONCLUSION: Within the limits of this study, the distribution of various calculus types was similar on different parts of the root surface; however, calculus was found more frequently on the coronal thirds than on the more apical regions. PMID- 15887507 TI - Risk stratification and dental management of the patient with cardiovascular diseases. Part II: Oral disease burden and principles of dental management. AB - Cardiovascular diseases are the leading cause of death in the United States and most other Western countries. In the United States alone, more than 1 million annual deaths and as many as three times that number of serious consequences can be attributed to these conditions. To provide care to patients with cardiovascular disease, oral health care providers must understand the disease, its treatment, and its impact on the patient's ability to undergo and respond to dental care. PMID- 15887508 TI - Periodontal status and sulcular Candida albicans colonization in patients with primary Sjogren's Syndrome. AB - OBJECTIVE: The purpose of this study was to compare the periodontal status and prevalence of sulcular Candida albicans between subjects with primary Sjogren's Syndrome (SS-1) and healthy control subjects. MATERIAL AND METHODS: Ten SS-1 subjects and 10 age- and sex-matched control subjects were recruited. All subjects met the comprehensive European Community Criteria for primary Sjogren's Syndrome. Periodontal probing depth (PD), clinical attachment loss (CAL), plaque and gingivitis were scored on index teeth in each subject. Gingival crevicular fluid (GCF) volume was measured from the same teeth using Periopaper strips. Candida albicans presence was determined by plating paper strips directly into culture media tubes (Oricult kit). RESULTS: Despite having similar mean PD measures and gingivitis and plaque scores, SS-1 subjects who had Sjogren's Syndrome for a mean of 8.8 years had significantly more CAL (5.4 mm vs 2.7 mm; P < .01) and GCF (101.3 +/- 4.25 microL vs 33.0 +/- 1.91 microL; P < .001) than healthy control subjects. Candida albicans was detected in the sulci of only one SS-1 subject and in none of the control subjects. CONCLUSION: These data indicate that the cohort of the SS-1 patients in this particular study have significantly more gingival recession and GCF than do control subjects. The increased attachment loss in SS-1 patients is not attributed to an increase in colonization of the gingival sulci by Candida albicans organisms. PMID- 15887509 TI - An intrusion injury as an example of interdisciplinary aspects in dental traumatology: a case report. AB - Traumatic injury to teeth presents a considerable challenge for the practitioner. Very often a variety of oral tissues are involved, such as enamel, dentin, periodontal ligament, alveolar bone, and mucosa. Extraoral lesions are often another focus of attention for the patient and the practitioner. To cover all these various demands and necessities in a proper and adequate manner, a sound knowledge and experience of many different dental fields is needed. This case report presents the treatment of an 8-year-old girl during which surgical, restorative, endodontic, and orthodontic practices cooperated to achieve an adequate rehabilitation of the patient. PMID- 15887510 TI - Mental foramen projected over the apex of the mandibular right second premolar. PMID- 15887511 TI - [Single channel recording with patch clamp method]. PMID- 15887512 TI - [Aging and psychological assessment of motor vehicle driving capability]. AB - BACKGROUND: The aim of the present project was to determine: (a) at what age the psychomotor functions begin to decline, which would justify routine testing to assess them, and (b) at what level the declining psychomotor functions can be recognized as a contraindication against driving motor vehicles. MATERIALS AND METHODS: The subjects were 186 motor vehicle drivers, aged 26-64 years. The parameters examined included eye-hand coordination and reaction time. RESULTS: The analysis of the results revealed that a significantly decreased level of psychomotor function could be noted at the age of 45 years. CONCLUSIONS: It was assumed that the results of psychological tests performed by persons with 0.05% BAC can be the empirical criterion for a decline that would significantly impair the driver's work capability. The findings of our study made it possible to develop a proposal for modifying the testing procedure and the principles of certifying work capability of motor vehicle drivers. PMID- 15887513 TI - [Effect of psychotherapy on autonomic balance in VDT-operators with somatoform disorders]. AB - BACKGROUND: The omnipresence of stress-inducing factors that stems from the permanent development of our civilization results in a constantly growing number of people with neurotic disorders. Computer technology, a very new labor technique, has become a stressful factor. Stressful factors initially affect the central nervous system, which modifies vegetative functions. The autonomic balance can be a measurable indicator of disease severity. MATERIALS AND METHODS: The study group consisted of 50 VDT-operators with neurotic disorders. Twenty five healthy VDT-operators formed the control group. The autonomic balance was investigated with Holter monitoring. Each of the 50 neurotic VDT-operators were qualified for a 3-month psychotherapy. RESULTS: The analysis of Holter recordings proved that the applied model of psychotherapy significantly reduced the number of sympathicotonics in favor of normotonics in the study group (p < 0.01). CONCLUSIONS: The Holter analysis can be considered as a reliable indicator of the effect of psychotherapy. PMID- 15887514 TI - [The level of anxiety, depression and aggression in nurses and their life and job satisfaction]. AB - BACKGROUND: In professions aimed at helping other people such as nursing, an increased level of anxiety, depression and aggression caused by extremely stressful work environment is always possible. It can be hypothesized that these emotions influence nurses' life and job satisfaction, through its decreasing or increasing. MATERIALS AND METHODS: In all, 102 nurses, employed in hospitals, outpatient clinics, hospices and old people's homes, were administered three questionnaires: 1) the Hospital Anxiety and Depression Scale-Modified; 2) the Life Satisfaction Scale; and 3) the job Satisfaction Scale. The survey was a closing part of the course on psychology carried out under the postgraduate education program. RESULTS: The statistical analysis of the results showed the increased level of anxiety in the youngest group of nurses, whereas the level of depression and aggression ranged from medium to low in the whole sample. The nurses' level of job satisfaction was higher than that of their life satisfaction and the differences were statistically significant in each of the three seniority categories. The evaluation of the relationship between life and job satisfaction and anxiety, depression, aggression and job seniority showed: (a) the effect of anxiety and depression, as self-contained factors, on life satisfaction, regardless of job seniority; (b) the effect of aggression interrelated with job seniority on job satisfaction. CONCLUSIONS: Anxiety is the major factor affecting life satisfaction; job satisfaction is less prone to the influence of negative emotions than life satisfaction. Bearing in mind a tendency shown by nurses to suppress their emotions as well as their stressful work environment, they should receive psychological support in its broadest sense. PMID- 15887515 TI - [Economic evaluation as a component in the evaluation of a prevention program]. AB - BACKGROUND: A growing gap between the required and available funds forces relevant bodies to use more effectively available financial means without decreasing the health care efficiency. Health policy makers have to take decisions on the kind, timing, and standard of medical care as well as on the mode and place of health service delivery. Moreover, they have to decide who should be provided with medical care under the National Health Fund. The economic evaluation techniques, primarily cost-benefit and cost-effect analyses, are essential tools supporting decision-making processes. MATERIALS AND METHODS: Under the LEONARDO DA VINCI (2000-2003) EUROPHAMILI project, an economic evaluation method has been developed and used to evaluate the antismoking program. In developing the method, the guidelines of the World Health Organization (WHO) were applied. Actual data on the cost of the program carried out in Lodz were used, and the benefits attained due to its implementation were estimated adopting the study assumptions based on actual data on the program efficiency and the literature data. RESULTS: In view of the expanded calculation model, only some examples of synthesized calculations are given. The results of the analysis show that the program financial profits exceed the costs incurred even if the assumed calculation parameters are changed. The performed calculations indicate that the shorter period of the prevention program implementation, the more advantageous cost-benefit relation. CONCLUSIONS: The performed economic evaluation helped to formulate prerequisites, which have to be met to facilitate the economic assessment of efficiency of the selected prevention programs. PMID- 15887516 TI - [Contact allergy to latex in health care workers--study of case reports]. AB - BACKGROUND: Contact allergy to latex is a relatively new problem that gives rise to much controversy among researchers. The aim of the study was to develop a study model of contact allergy induced by latex proteins. MATERIALS AND METHODS: The study covered 60 health care workers who had reported decreased tolerance of rubber gloves. They were subjected to routine dermatological examinations, patch tests to a standard set of allergens (Chemotechnique Diagnosis, Sweden), expanded to include high ammoniac latex, and prick tests with latex and common inhalatory allergens. In addition, total and latex specific IgE antibodies were determined. RESULTS: Of the total group, contact allergy to latex was found in 5 persons, including 3 persons with concurrent immediate reaction to latex, and 2 persons with allergy of the isolated nature. Only in 1 person coexistent late allergy to latex and rubber accelerator of the thiurams group were observed. CONCLUSIONS: Path tests with 30% ammoniac latex seem to be a useful diagnostic method in the detection of contact allergy to latex. In case of doubtful reactions, the use of lower allergen concentrations is recommended. PMID- 15887517 TI - [Determination of fumes and their elements from flux cored arc welding]. AB - BACKGROUND: The aim of this work was to assay the concentration levels and composition of welding fumes, released during flux cored arc welding, to assess exposure of welders. MATERIALS AND METHODS: Concentrations and welding fume components, such as iron, manganese, chromium (including the soluble and chromium VI), nickel, copper, calcium, aluminium, barium, and fluorides (including hydrogen fluoride) were determined in the air of six industrial plants (shipyards, mechanical engineering plants and a power station) at the breathing zones of the welders who used 10 types of wires during flux cored arc welding. The following determination methods were used: gravimetry (fumes), AAS (metals), and spectrophotometry (chromium VI, fluorides--including hydrogen fluoride). RESULTS: The results made it possible to determine the relationship between concentrations of welding fume and its elements, and to assess worker's exposure. Time weighted average concentrations of the welding fumes and its elements at the worker's breathing zone were: mg/m3: dust 0.2-24.3; Fe 0.2-6.7; Mn 0.01-1.8; Cr 0.004-0.5 (mainly Cr III); Ca 0.004-2.5; Ni < or = 0.004; Cu < 0.002-0.05; Al < 0.14-0.4; Ba < 0.14; F- 0.07-0.43. CONCLUSIONS: The welders using some types of flux cored welding wires worked in conditions harmful to their health owing to the considerably exceeded TLV value for fume and MAC values for manganese, and occasional slightly excessive MAC values for calcium and iron. PMID- 15887518 TI - [Justification for screening laboratory tests not related to occupational exposure ordered by occupational medicine physicians: an analytical study]. AB - The aim of the study was to analyze whether laboratory tests not related with occupational exposure ordered by occupational medicine physicians are justified. In Poland, prophylactic standards in occupational medicine are determined by law and the minimum range of laboratory tests and consultations by specialists is laid down. The extension of its range to include screening in all workers is disputable. The authors analyze the usefulness of the most common laboratory tests performed in screening (blood count, erythrocyte sedimentation rate, lipid profile, glucose in blood, urine). In part of those examinations, the benefit to a patient, e.g., the detection of a pathology, is questionable if they are not supported by a thorough analysis of the patient's state of health and risk factors. PMID- 15887519 TI - [Toxic effect of dust and fumes of aluminium and its compounds on workers' respiratory tract]. AB - Based on the literature review, the authors discuss problems concerning differentiated exposure of workers to dust and fumes of aluminum and its compounds and describe the observed toxic effect on the respiratory tract. Long- term occupational exposure to the above factors leads to changes in lungs of the pneumoconiotic nature. Other disorders presented in the literature include: pulmonary fibrosis, pulmonary alveolitis and alveolar proteinosis, asthma, chronic bronchitis, and chronic pneumonia. The respiratory effect depends to some extent on the form of aluminum or the stage of processing in which exposure occurs. Numerous studies of workers occupationally exposed to aluminum dust and fumes have demonstrated the increase in the incidence of pulmonary fibrosis, depending on the air concentration of respirable fraction of dust. PMID- 15887520 TI - [Glutathione: its biosynthesis, induction agents and concentrations in selected diseases]. AB - Glutathione (GSH) is in a constant state of metabolic turnover. Because it is actively synthesized, it also must be degraded. In the first step of GSH synthesis, an amide linkage is formed between cysteine and glutamate catalyzed by gamma-glutamylcysteine synthetase. GSH synthetase catalyzes the reaction between amine residue of glycine and the cysteine carboxyl from gamma-glutamylcysteine dipeptide to form GSH. GSH is transported out of the cell and degraded by the membrane-bound enzyme gammaGT, which removes the gamma-glutamyl moiety, and by dipeptidases, which remove the glycine moiety. Glutathione is present in most of the plants and animals' tissues that constitute human diet. Thiol redox cycles play central roles in the antioxidant defense network. Lipoate and vitamins and other reducing factors affect the increase in glutathione concentrations in cells by the rise of the concentrations of reduced cysteine. The level of GSH in humans may be increased by taking different glutathione monoester (drug) or factors reducing cystyne to cysteine and increasing availability of this amino acid to GSH synthesis. GSH plays a critical role in cellular mechanisms that lead to cell death. The cancer cells resistant to apoptosis have higher intracellular GSH levels. The fact that numerous diseases are induced by RFT (that cause glutathione depletion) it seems that an in-depth study of the dietetic and pharmacological manners of manipulation of the GSH amount and availability may become in future a tool of great importance in the prevention of many illnesses. PMID- 15887521 TI - [Problems of diagnostic tests for sanitary and epidemiologic purposes]. PMID- 15887522 TI - [The MATRA project on "The redefinition of the responsibility of the Minister of Health for health care of working people in Poland"]. PMID- 15887523 TI - [Summary report on an international scientific conference "Towards multi dimensional approach in occupational health service: scientific evidence, social consensus, human values", Modena, Italy, on 13-16 October 2004]. PMID- 15887524 TI - [Summary report on the 10th international seminar on ergonomics and work safety and hygiene in agriculture "Contamination of working and living environments in agricultural regions", Lublin, Poland, on 18-20 October 2004]. PMID- 15887525 TI - Estimation of the cortical connectivity by high-resolution EEG and structural equation modeling: simulations and application to finger tapping data. AB - Today, the concept of brain connectivity plays a central role in the neuroscience. While functional connectivity is defined as the temporal coherence between the activities of different brain areas, the effective connectivity is defined as the simplest brain circuit that would produce the same temporal relationship as observed experimentally between cortical sites. The most used method to estimate effective connectivity in neuroscience is the structural equation modeling (SEM), typically used on data related to the brain hemodynamic behavior. However, the use of hemodynamic measures limits the temporal resolution on which the brain process can be followed. The present research proposes the use of the SEM approach on the cortical waveforms estimated from the high-resolution EEG data, which exhibits a good spatial resolution and a higher temporal resolution than hemodynamic measures. We performed a simulation study, in which different main factors were systematically manipulated in the generation of test signals, and the errors in the estimated connectivity were evaluated by the analysis of variance (ANOVA). Such factors were the signal-to-noise ratio and the duration of the simulated cortical activity. Since SEM technique is based on the use of a model formulated on the basis of anatomical and physiological constraints, different experimental conditions were analyzed, in order to evaluate the effect of errors made in the a priori model formulation on its performances. The feasibility of the proposed approach has been shown in a human study using high-resolution EEG recordings related to finger tapping movements. PMID- 15887526 TI - A mathematical analysis of SFAP convolutional models. AB - In this paper we compare, from a mathematical point of view, two well-recognized single fiber action potential (SFAP) convolutional models: the Nandedkar-Stalberg (N-S) model and the Dimitrov-Dimitrova (D-D) model. Junction waves appear in N-S SFAPs due to the onset and extinction of the monopoles whereas in D-D SFAPs these waves appear only when the dipoles reach the fiber/tendon junctions. D-D junction waves model more accurately the out-of-the-main-spike waveforms that appear in experimental SFAPs. The origin of junction waves lies in the discontinuities of the impulse responses. There are two kinds of these waves caused by the two types of existing discontinuities (in the impulse response function and in its derivative). We model each kind of discontinuity with a different mathematical function. Using these functions, the N-S and D-D impulse responses can be split and, therefore, the junction waves can be separated from the spike component of the SFAP. The expansion of the impulse response helps us to understand the differences between the N-S and D-D junction waves. PMID- 15887527 TI - Dynamic HIV/AIDS parameter estimation with application to a vaccine readiness study in southern Africa. AB - This paper proposes a procedure of parameter estimation for all parameters of the three-dimensional HIV model. The least square based procedure uses standard optimization routines to allow parameter extraction for individual patients. It is shown how additional information from outside a measurement dataset can be included in the estimation routine to increase the reliability and accuracy of parameter estimates. A dataset from 44 patients of Southern Africa is analyzed to find the set point and the time until set point for these patients together with an estimate of the model parameters with confidence intervals for the cohort. The procedure is also applied to a long-term dataset of the HIV/AIDS progression to find possible variations in parameters. PMID- 15887528 TI - A network of electronic neural oscillators reproduces the dynamics of the periodically forced pyloric pacemaker group. AB - Low-dimensional oscillators are a valuable model for the neuronal activity of isolated neurons. When coupled, the self-sustained oscillations of individual free oscillators are replaced by a collective network dynamics. Here, dynamical features of such a network, consisting of three electronic implementations of the Hindmarsh-Rose mathematical model of bursting neurons, are compared to those of a biological neural motor system, specifically the pyloric CPG of the crustacean stomatogastric nervous system. We demonstrate that the network of electronic neurons exhibits realistic synchronized bursting behavior comparable to the biological system. Dynamical properties were analyzed by injecting sinusoidal currents into one of the oscillators. The temporal bursting structure of the electronic neurons in response to periodic stimulation is shown to bear a remarkable resemblance to that observed in the corresponding biological network. These findings provide strong evidence that coupled nonlinear oscillators realistically reproduce the network dynamics experimentally observed in assemblies of several neurons. PMID- 15887529 TI - Simulation of ST segment changes during subendocardial ischemia using a realistic 3-D cardiac geometry. AB - The mechanisms underlying the ST segment shifts associated with subendocardial ischemia remain unclear. The aim of this paper is to shed further light on the subject through numerical simulations of these shifts. A realistic three dimensional model of the ventricles, including fiber rotation and anisotropy, is embedded in a nonhomogeneous torso model. A simplification of the bidomain model is used to calculate only the ST segment shift, assuming known values of the transmembrane potential during the plateau and rest phases. A similar simulation is performed in two dimensions. The simulation results suggest that subendocardial ischemia can be located by ST segment shift on the epicardial and torso surfaces. It is shown that ST elevation is associated with the transmural ischemic boundary, while ST depression is associated with the lateral ischemic boundaries. PMID- 15887530 TI - Estimating the growth kinetics of experimental tumors from as few as two determinations of tumor size: implications for clinical oncology. AB - Clinical information on tumor growth is often limited to a few determinations of the size of the tumor burden taken at variable time. As a consequence, fitting of growth equations to clinical data is hampered by the small number of available data. On the other hand, characterising the tumor growth kinetics in terms of clinically relevant parameters, such as the doubling time of the tumors, is increasingly required to optimize and personalise treatments. A computational method is presented which can estimate the growth kinetics of tumors from as few as two determinations of its size taken at two successive time points, provided the size at which tumor growth saturates is known. The method is studied by using experimental data obtained in vitro with multicell tumor spheroids and in vivo with tumors grown in mice, and its outputs are compared to those obtained by fitting of experimental data with the Gompertz growth equation. Under certain assumptions and limitations the method provides comparable estimates of the doubling time of tumors with respect to the classical nonlinear fitting approach. The method is then tested against simulated tumor growth trajectories spanning the range of tumor sizes observed in the clinics. The simulations show that a relative classification of tumors on the basis of their growth kinetics can be obtained even if the size at which tumor growth saturates is not known. This result opens the possibility to classify patients bearing fast or slow growing tumors and, hence, to adapt therapeutic regimens under a more rationale basis. PMID- 15887531 TI - Sequential finite element model of tissue electropermeabilization. AB - Permeabilization, when observed on a tissue level, is a dynamic process resulting from changes in membrane permeability when exposing biological cells to external electric field (E). In this paper we present a sequential finite element model of E distribution in tissue which considers local changes in tissue conductivity due to permeabilization. These changes affect the pattern of the field distribution during the high voltage pulse application. The presented model consists of a sequence of static models (steps), which describe E distribution at discrete time intervals during tissue permeabilization and in this way present the dynamics of electropermeabilization. The tissue conductivity for each static model in a sequence is determined based on E distribution from the previous step by considering a sigmoid dependency between specific conductivity and E intensity. Such a dependency was determined by parameter estimation on a set of current measurements, obtained by in vivo experiments. Another set of measurements was used for model validation. All experiments were performed on rabbit liver tissue with inserted needle electrodes. Model validation was carried out in four different ways: 1) by comparing reversibly permeabilized tissue computed by the model and the reversibly permeabilized area of tissue as obtained in the experiments; 2) by comparing the area of irreversibly permeabilized tissue computed by the model and the area where tissue necrosis was observed in experiments; 3) through the comparison of total current at the end of pulse and computed current in the last step of sequential electropermeabilization model; 4) by comparing total current during the first pulse and current computed in consecutive steps of a modeling sequence. The presented permeabilization model presents the first approach of describing the course of permeabilization on tissue level. Despite some approximations (ohmic tissue behavior) the model can predict the permeabilized volume of tissue, when exposed to electrical treatment. Therefore, the most important contribution and novelty of the model is its potentiality to be used as a tool for determining parameters for effective tissue permeabilization. PMID- 15887532 TI - Support vector machines for automated gait classification. AB - Ageing influences gait patterns causing constant threats to control of locomotor balance. Automated recognition of gait changes has many advantages including, early identification of at-risk gait and monitoring the progress of treatment outcomes. In this paper, we apply an artificial intelligence technique [support vector machines (SVM)] for the automatic recognition of young-old gait types from their respective gait-patterns. Minimum foot clearance (MFC) data of 30 young and 28 elderly participants were analyzed using a PEAK-2D motion analysis system during a 20-min continuous walk on a treadmill at self-selected walking speed. Gait features extracted from individual MFC histogram-plot and Poincare-plot images were used to train the SVM. Cross-validation test results indicate that the generalization performance of the SVM was on average 83.3% (+/-2.9) to recognize young and elderly gait patterns, compared to a neural network's accuracy of 75.0+/-5.0%. A "hill-climbing" feature selection algorithm demonstrated that a small subset (3-5) of gait features extracted from MFC plots could differentiate the gait patterns with 90% accuracy. Performance of the gait classifier was evaluated using areas under the receiver operating characteristic plots. Improved performance of the classifier was evident when trained with reduced number of selected good features and with radial basis function kernel. These results suggest that SVMs can function as an efficient gait classifier for recognition of young and elderly gait patterns, and has the potential for wider applications in gait identification for falls-risk minimization in the elderly. PMID- 15887533 TI - Line-source modeling and estimation with magnetoencephalography. AB - We propose a number of source models that are spatially distributed on a line for magnetoencephalography (MEG) using both a spherical head with radial sensors for more efficient computation and a realistic head model for more accurate results. We develop these models with increasing degrees of freedom, derive forward solutions, maximum-likelihood (ML) estimates, and Cramer-Rao bound (CRB) expressions for the unknown source parameters. A model selection method is applied to select the most appropriate model. We also present numerical examples to compare the performances and computational costs of the different models, to determine the regions where better estimates are possible and when it is possible to distinguish between line and focal sources. We demonstrate the usefulness of the proposed line-source models over the previously available focal source model in certain distributed source cases. Finally, we apply our methods to real MEG data, the N2O response after electric stimulation of the median nerve known to be an extended source. PMID- 15887534 TI - Estimation of coherence between blood flow and spontaneous EEG activity in neonates. AB - Blood flow to the brain responds to changes in neuronal activity and, thus, metabolic demand. In earlier work, we observed correlation between cerebral blood flow and spontaneous electroencephalogram (EEG) activity in neonates. Using coherence, we now found that during Trace Alternant EEG activity in quiet sleep of normal term neonates, this correlation is strongest at frequencies around 0.1 Hz, reaching statistical significance (p < 0.05) in six of the nine subjects studied (p < 0.07 in eight subjects). Due to noise, artifact, and spontaneous changes in the subjects' EEG patterns, the signals investigated included epochs of missing samples. We, therefore, developed a novel algorithm for the estimation of coherence in such data and applied a Monte Carlo (surrogate data) method for its statistical analysis. This process provides a test for the statistical significance of the maximum coherence within a selected frequency band. In addition to permitting further insight into the mechanisms of cerebral blood flow control, these algorithms are potentially of great benefit in a wide range of biomedical applications, where interrupted (gapped) recordings are often a problem. PMID- 15887535 TI - Application of cross time-frequency analysis to postural sway behavior: the effects of aging and visual systems. AB - In this paper, the effects of visual feedback and aging on postural sway systems and signals are investigated by analyzing the transient phase difference between "input" and "output" which correspond to center of pressure (COP) and center of mass (COM), respectively. In order to analyze the transient phase difference characteristics of COP and COM, a relatively new cross time-frequency analysis technique that provides time- and frequency-localized phase difference information is utilized. The feedback control process in the postural sway is interpreted in terms of a feedback compensator which is characterized in terms of a phase difference. Using the experimental results of the transient phase difference obtained from the cross time-frequency distribution, it is demonstrated that the postural control of young persons are more stable and rely more on visual sensory feedback to stabilize postural control compared to that of the elderly persons. PMID- 15887536 TI - Estimation of M-wave scale factor during sustained contractions at high stimulation rate. AB - In this paper, we propose a time-domain index to assess M-wave widening during high-frequency stimulation, as an objective parameter for quantifying muscle fatigue. At high stimulation frequencies, signal truncation, due to the delivery of the electrical stimulus before the M-wave generated by the previous stimulus extinguishes, biases the spectral frequency variables usually computed to estimate M-wave widening. Thus, we propose an estimator of the scale factor between two truncated M-waves. The estimator is derived from the Scale Transforms of the two signals, with an efficient implementation that avoids limits of resolution. The method was tested on both simulated and experimental signals. The simulations showed that the proposed technique is significantly less affected by signal truncation than previous approaches. The experimental recordings were collected from 11 subjects at stimulation frequencies of 20, 40, and 60 Hz. The scale factor estimation assessed M-wave widening in the three conditions, differentiating between the different rates of change of signal widening. The method proved to be significantly superior to M-wave spectral analysis. The technique can be applied to investigate myoelectric manifestations of muscle fatigue at stimulation rates that could not be analyzed in the past and, thus, opens new perspectives in the evaluation of electrical stimulation for training and rehabilitation protocols. PMID- 15887537 TI - Orthonormal-basis partitioning and time-frequency representation of cardiac rhythm dynamics. AB - Although a number of time-frequency representations have been proposed for the estimation of time-dependent spectra, the time-frequency analysis of multicomponent physiological signals, such as beat-to-beat variations of cardiac rhythm or heart rate variability (HRV), is difficult. We thus propose a simple method for 1) detecting both abrupt and slow changes in the structure of the HRV signal, 2) segmenting the nonstationary signal into the less nonstationary portions, and 3) exposing characteristic patterns of the changes in the time frequency plane. The method, referred to as orthonormal-basis partitioning and time-frequency representation (OPTR), is validated using simulated signals and actual HRV data. Here we show that OPTR can be applied to long multicomponent ambulatory signals to obtain the signal representation along with its time varying spectrum. PMID- 15887538 TI - Wavelet-based compression of M-FISH images. AB - Multiplex fluorescence in situ hybridization (M-FISH) is a recently developed technology that enables multi-color chromosome karyotyping for molecular cytogenetic analysis. Each M-FISH image set consists of a number of aligned images of the same chromosome specimen captured at different optical wavelength. This paper presents embedded M-FISH image coding (EMIC), where the foreground objects/chromosomes and the background objects/images are coded separately. We first apply critically sampled integer wavelet transforms to both the foreground and the background. We then use object-based bit-plane coding to compress each object and generate separate embedded bitstreams that allow continuous lossy-to lossless compression of the foreground and the background. For efficient arithmetic coding of bit planes, we propose a method of designing an optimal context model that specifically exploits the statistical characteristics of M FISH images in the wavelet domain. Our experiments show that EMIC achieves nearly twice as much compression as Lempel-Ziv-Welch coding. EMIC also performs much better than JPEG-LS and JPEG-2000 for lossless coding. The lossy performance of EMIC is significantly better than that of coding each M-FISH image with JPEG 2000. PMID- 15887539 TI - Efficient electromagnetic source imaging with adaptive standardized LORETA/FOCUSS. AB - Functional brain imaging and source localization based on the scalp's potential field require a solution to an ill-posed inverse problem with many solutions. This makes it necessary to incorporate a priori knowledge in order to select a particular solution. A computational challenge for some subject-specific head models is that many inverse algorithms require a comprehensive sampling of the candidate source space at the desired resolution. In this study, we present an algorithm that can accurately reconstruct details of localized source activity from a sparse sampling of the candidate source space. Forward computations are minimized through an adaptive procedure that increases source resolution as the spatial extent is reduced. With this algorithm, we were able to compute inverses using only 6% to 11% of the full resolution lead-field, with a localization accuracy that was not significantly different than an exhaustive search through a fully-sampled source space. The technique is, therefore, applicable for use with anatomically-realistic, subject-specific forward models for applications with spatially concentrated source activity. PMID- 15887540 TI - Hollow metal microneedles for insulin delivery to diabetic rats. AB - The goal of this study was to design, fabricate, and test arrays of hollow microneedles for minimally invasive and continuous delivery of insulin in vivo. As a simple, robust fabrication method suitable for inexpensive mass production, we developed a modified-LIGA process to micromachine molds out of polyethylene terephthalate using an ultraviolet laser, coated those molds with nickel by electrodepostion onto a sputter-deposited seed layer, and released the resulting metal microneedle arrays by selectively etching the polymer mold. Mechanical testing showed that these microneedles were sufficiently strong to pierce living skin without breaking. Arrays containing 16 microneedles measuring 500 microm in length with a 75 microm tip diameter were then inserted into the skin of anesthetized, diabetic, hairless rats. Insulin delivery through microneedles caused blood glucose levels to drop steadily to 47% of pretreatment values over a 4-h insulin delivery period and were then approximately constant over a 4-h postdelivery monitoring period. Direct measurement of plasma insulin levels showed a peak value of 0.43 ng/ml. Together, these data suggest that microneedles can be fabricated and used for in vivo insulin delivery. PMID- 15887541 TI - Stochastic frequency signature for chemical sensing using noninvasive neuronelectronic interface. AB - The detection of chemical agents is important in many areas including environmental pollutants, toxins, biological and chemical pollutants. As "smart" cells, with strong information encoding ability, neurons can be treated as independent sensing elements. A hybrid circuit of a semiconductor chip with dissociated neurons formed both sensors and transducers. Stochastic frequency spectrum was used to differentiate a mixture of chemical agents with effect on the opening of different ion channels. The frequency of spike trains revealed the concentration of the chemical agent, where the characteristic tuning curve revealed the identity. "Fatigue" experiment was performed to explore the "refreshing" ability and "memory" effect of neurons by cyclic and cascaded sensing. "Neuronelectronic noses" such as this should have wide potential applications, most notably in environmental and medical monitoring. PMID- 15887542 TI - Multielectrode microprobes for deep-brain stimulation fabricated with a customizable 3-D electroplating process. AB - Although deep-brain stimulation (DBS) can be used to improve some of the severe symptoms of Parkinson's disease (e.g., Bradykinesia, rigidity, and tremors), the mechanisms by which the symptoms are eliminated are not well understood. Moreover, DBS does not prevent neurodegeneration that leads to dementia or death. In order to fully investigate DBS and to optimize its use, a comprehensive long term stimulation study in an animal model is needed. However, since the brain region that must be stimulated, known as the subthalamic nucleus (STN), is extremely small (500 microm x 500 microm x 1 mm) and deep within the rat brain (10 mm), the stimulating probe must have geometric and mechanical properties that allow accurate positioning in the brain, while minimizing tissue damage. We have designed, fabricated, and tested a novel micromachined probe that is able to accurately stimulate the STN. The probe is designed to minimize damage to the surrounding tissue. The probe shank is coated with gold and the electrode interconnects are insulated with silicon nitride for biocompatibility. The probe has four platinum electrodes to provide a variety of spatially distributed stimuli, and is formed in a novel 3-D plating process that results in a microwire like geometry (i.e., smoothly tapering diameter) with a corresponding mechanically stable shank. PMID- 15887543 TI - Motion artifact cancellation in NIR spectroscopy using Wiener filtering. AB - We present a Wiener filtering based algorithm for the elimination of motion artifacts present in Near Infrared (NIR) spectroscopy measurements. Until now, adaptive filtering was the only technique used in the noise cancellation in NIR studies. The results in this preliminary study revealed that the proposed method gives better estimates than the classical adaptive filtering approach without the need for additional sensor measurements. Moreover, this novel technique has the potential to filter out motion artifacts in functional near infrared (fNIR) signals, too. PMID- 15887544 TI - A 1.48-mW low-phase-noise analog frequency modulator for wireless biotelemetry. AB - This paper presents a low-phase-noise, hybrid LC-tank, analog frequency modulator for wireless biotelemetry employing on-chip NMOS varactors in the inversion region as the frequency tuning element. We demonstrate that a correct estimate for the destination signal-to-noise ratio, which quantifies the quality of the wirelessly received signal in a frequency-modulated biotelemetry system, is only achieved after taking into account the large-signal oscillation effect on the tank varactor. A prototype chip is fabricated using AMI 1.5-microm double-poly double-metal n-well CMOS process, and exhibits a measured gain factor of 1.21 MHz/V in the mid-range of the tuning voltage and a phase noise of -88.6 dBc/Hz at 10-kHz offset from the 95.1-MHz carrier while dissipating 1.48 mW from a 3 V power supply leading to a figure of merit (FOM) of -166.5 dBc/Hz. The VCO is successfully interfaced with a penetrating silicon microelectrode with 700 microm2 iridium recording sites for wireless in vitro recording of a 50 Hz simulated normal sinus rhythm signal from saline over a distance of approximately 0.25 m. Given a typical gain of approximately 40 dB for fully integrated front end bioamplifiers, a wireless recording microsystem employing this VCO would be capable of detecting input biopotentials down to the submilivolt range. PMID- 15887545 TI - Performance evaluation of functional medical imaging compression via optimal sampling schedule designs and cluster analysis. AB - In previous work we have described a technique for the compression of positron emission tomography (PET) image data in the spatial and temporal domains based on optimal sampling schedule designs (OSS) and cluster analysis. It can potentially achieve a high data compression ratio greater than 80:1. However, the number of distinguishable cluster groups in dynamic PET image data is a critical issue for this algorithm that has not been experimentally analyzed on clinical data. In this paper, the problem of experimentally determining the ideal cluster number for the algorithm for PET brain data is addressed. PMID- 15887546 TI - An input-delay neural-network-based approach for piecewise ECG signal compression. AB - We propose an input delay neural network (IDNN) based time series prediction algorithm for compressing electrocardiogram (ECG) signals. Our algorithm has been tested and successfully compared vis-a-vis other popular techniques for its compression efficiency and reconstruction capability. PMID- 15887547 TI - Modeling and characterization of a valved glaucoma drainage device with implications for enhanced therapeutic efficacy. AB - We report on modeling and bench test results targeted at better understanding of valved glaucoma drainage devices (GDDs), a common current surgical treatment for glaucoma. A simple equivalent circuit is described to model fluid mechanical behavior of the aqueous humor in an eye with glaucoma, both before and after implantation of a valved GDD. Finite element method simulations (FEM), based on the lubrication-von Karman model, are then performed to analyze the valve's mechanical and fluidic performance. Using nanoporous membranes to mimic the in vivo fibrous capsule, we have developed a microfluidic bench test to simulate the aqueous humor flow and the post-implantation fibrous tissue encapsulation around the GDD back plate. Our numerical and bench test results show that, contrary to the prevailing belief, the valve significantly contributes to the total pressure drop even after fibrous capsule formation. Furthermore, we show that bypassing the valve through a simple polyimide tube insertion will dramatically lower the intraocular pressure (IOP) after fibrous capsule formation. This may offer a new treatment option in some patients with advanced glaucoma. PMID- 15887548 TI - Easy-to-prepare assembly array of Tungsten microelectrodes. AB - This paper provides a detailed process flow for fabricating an easy-to-prepare, inexpensive, dense array of tungsten microelectrodes. We designed the process flow to minimize routine tasks by separating an initial preparation of a master mold from a routine preparation of substrate replication, array assembly and tip processing. Sandblast processing first produced a glass mold with a pattern of a series of protruding lines at a requested interval of a few hundred micrometers. Copying the groove pattern onto polystyrene mass-produced a replica substrate. Tungsten probes were then aligned on the substrate, and the tips of probes were finely processed in the block. PMID- 15887549 TI - Multiple time-varying dynamic analysis using multiple sets of basis functions. AB - We extend a recently developed algorithm that expands the time-varying parameters onto a single set of basis functions, to multiple sets of basis functions. This feature allows the capability to capture many different dynamics that may be inherent in the system. A single set of basis functions that has its own unique characteristics can best capture dynamics of the system that have similar features. Therefore, for systems that have multiple dynamics, the use of a single set of basis functions may not be adequate. Computer simulation examples do indeed show the benefit of using multiple sets of basis functions over the single set of basis functions for cases with many switching dynamics. Moreover, the proposed method remains accurate even under significant noise contamination. Application of the proposed approach to blood pressure data likewise indicate better tracking capability of the two sets of basis function than the recursive least squares or a single set of basis functions. PMID- 15887550 TI - 3-D deformable image registration: a topology preservation scheme based on hierarchical deformation models and interval analysis optimization. AB - This paper deals with topology preservation in three-dimensional (3-D) deformable image registration. This work is a nontrivial extension of, which addresses the case of two-dimensional (2-D) topology preserving mappings. In both cases, the deformation map is modeled as a hierarchical displacement field, decomposed on a multiresolution B-spline basis. Topology preservation is enforced by controlling the Jacobian of the transformation. Finding the optimal displacement parameters amounts to solving a constrained optimization problem: The residual energy between the target image and the deformed source image is minimized under constraints on the Jacobian. Unlike the 2-D case, in which simple linear constraints are derived, the 3-D B-spline-based deformable mapping yields a difficult (until now, unsolved) optimization problem. In this paper, we tackle the problem by resorting to interval analysis optimization techniques. Care is taken to keep the computational burden as low as possible. Results on multipatient 3-D MRI registration illustrate the ability of the method to preserve topology on the continuous image domain. PMID- 15887551 TI - Specification of the observation model for regularized image up-sampling. AB - Regularization is one of the most promising methods for image up-sampling, which is an ill-posed inverse problem. A key element of such a regularization approach is the observation model relating the observed lower resolution (LR) image to the desired higher resolution (HR) up-sampled image, used in the data-fidelity term of the regularization cost function. This paper presents an algorithm to determine this observation model based on a model of the physical acquisition process for the LR image, and the ideal acquisition process for the desired HR image, both from the same underlying continuous image. The method is illustrated with typical scenarios corresponding to LR and HR cameras modeled by either Gaussian or rectangular apertures. Experiments with some regularized image up samplers demonstrate the importance of using the correct, adapted observation model as determined by our algorithm. Index Terms-Camera aperture, data fidelity, image up-sampling, interpolation, multidimensional signal processing, observation model, power spectral density (PSD), super-resolution. PMID- 15887552 TI - Multiframe selective information fusion from robust error estimation theory. AB - A dynamic procedure for selective information fusion from multiple image frames is derived from robust error estimation theory. The fusion rate is driven by the anisotropic gain function, defined to be the difference between the Gaussian smoothed-edge maps of a given input frame and of an evolving synthetic output frame. The gain function achieves both selection and rapid fusion of relatively sharper features from each input frame compared to the synthetic frame. Effective applications are demonstrated for image sharpening in imaging through atmospheric turbulence, for multispectral fusion of the RGB spectral components of a scene, for removal of blurred visual obstructions from in front of a distant focused scene, and for high-resolution two-dimensional display of three-dimensional objects in microscopy. PMID- 15887553 TI - Adaptive estimation of normals and surface area for discrete 3-D objects: application to snow binary data from X-ray tomography. AB - Estimating the normal vector field on the boundary of discrete three-dimensional objects is essential for rendering and image measurement problems. Most of the existing algorithms do not provide an accurate determination of the normal vector field for shapes that present edges. Here, we propose a new and simple computational method in order to obtain accurate results on all types of shapes, whatever their local convexity degree. The presented method is based on the gradient vector field analysis of the object distance map. This vector field is adaptively filtered around each surface voxel using angle and symmetry criteria so that as many relevant contributions as possible are accounted for. This optimizes the smoothing of digitization effects while preserving relevant details of the processed numerical object. Thanks to the precise normal field obtained, a projection method can be proposed to immediately derive the surface area from a raw discrete object. An empirical justification of the validity of such an algorithm in the continuous limit is also provided. Some results on simulated data and snow images from X-ray tomography are presented, compared to the Marching Cubes and Convex Hull results, and discussed. PMID- 15887554 TI - New adaptive partial distortion search using clustered pixel matching error characteristic. AB - In order to reduce the computation load, many conventional fast block-matching algorithms have been developed to reduce the set of possible searching points in the search window. All of these algorithms produce some quality degradation of a predicted image. Alternatively, another kind of fast block-matching algorithms which do not introduce any prediction error as compared with the full-search algorithm is to reduce the number of necessary matching evaluations for every searching point in the search window. The partial distortion search (PDS) is a well-known technique of the second kind of algorithms. In the literature, many researches tried to improve both lossy and lossless block-matching algorithms by making use of an assumption that pixels with larger gradient magnitudes have larger matching errors on average. Based on a simple analysis, it is found that, on average, pixel matching errors with similar magnitudes tend to appear in clusters for natural video sequences. By using this clustering characteristic, we propose an adaptive PDS algorithm which significantly improves the computation efficiency of the original PDS. This approach is much better than other algorithms which make use of the pixel gradients. Furthermore, the proposed algorithm is most suitable for motion estimation of both opaque and boundary macroblocks of an arbitrary-shaped object in MPEG-4 coding. PMID- 15887555 TI - Variational optical flow computation in real time. AB - This paper investigates the usefulness of bidirectional multigrid methods for variational optical flow computations. Although these numerical schemes are among the fastest methods for solving equation systems, they are rarely applied in the field of computer vision. We demonstrate how to employ those numerical methods for the treatment of variational optical flow formulations and show that the efficiency of this approach even allows for real-time performance on standard PCs. As a representative for variational optic flow methods, we consider the recently introduced combined local-global method. It can be considered as a noise robust generalization of the Horn and Schunck technique. We present a decoupled, as well as a coupled, version of the classical Gauss-Seidel solver, and we develop several multgrid implementations based on a discretization coarse grid approximation. In contrast, with standard bidirectional multigrid algorithms, we take advantage of intergrid transfer operators that allow for nondyadic grid hierarchies. As a consequence, no restrictions concerning the image size or the number of traversed levels have to be imposed. In the experimental section, we juxtapose the developed multigrid schemes and demonstrate their superior performance when compared to unidirectional multgrid methods and nonhierachical solvers. For the well-known 316 x 252 Yosemite sequence, we succeeded in computing the complete set of dense flow fields in three quarters of a second on a 3.06-GHz Pentium4 PC. This corresponds to a frame rate of 18 flow fields per second which outperforms the widely-used Gauss-Seidel method by almost three orders of magnitude. PMID- 15887556 TI - An optimal nonlinear extension of linear filters based on distributed arithmetic. AB - Distributed arithmetic (DA)-based implementation of linear filters relies on the linear nature of this operation and has been suggested as a multiplication free solution. In this work, we introduce a nonlinear extension of linear filters optimizing under mean-square error criterion the memory function [(MF) multivariate Boolean function with not only binary output] which is in the core of DA based implementation. Such an extension will improve the filtering of noise which may contain non-Gaussian components without increasing the complexity of implementation. Experiments on real images have shown the superiority of the proposed filters over the optimal linear filters. Different versions of these filters are also considered for an impulsive noise removal, faster processing, and filtering using large input data windows. PMID- 15887557 TI - A soft double regularization approach to parametric blind image deconvolution. AB - This paper proposes a blind image deconvolution scheme based on soft integration of parametric blur structures. Conventional blind image deconvolution methods encounter a difficult dilemma of either imposing stringent and inflexible preconditions on the problem formulation or experiencing poor restoration results due to lack of information. This paper attempts to address this issue by assessing the relevance of parametric blur information, and incorporating the knowledge into the parametric double regularization (PDR) scheme. The PDR method assumes that the actual blur satisfies up to a certain degree of parametric structure, as there are many well-known parametric blurs in practical applications. Further, it can be tailored flexibly to include other blur types if some prior parametric knowledge of the blur is available. A manifold soft parametric modeling technique is proposed to generate the blur manifolds, and estimate the fuzzy blur structure. The PDR scheme involves the development of the meaningful cost function, the estimation of blur support and structure, and the optimization of the cost function. Experimental results show that it is effective in restoring degraded images under different environments. PMID- 15887558 TI - Landcover classification in MRF context using Dempster-Shafer fusion for multisensor imagery. AB - This work deals with multisensor data fusion to obtain landcover classification. The role of feature-level fusion using the Dempster-Shafer rule and that of data level fusion in the MRF context is studied in this paper to obtain an optimally segmented image. Subsequently, segments are validated and classification accuracy for the test data is evaluated. Two examples of data fusion of optical images and a synthetic aperture radar image are presented, each set having been acquired on different dates. Classification accuracies of the technique proposed are compared with those of some recent techniques in literature for the same image data. PMID- 15887559 TI - Forensic analysis of nonlinear collusion attacks for multimedia fingerprinting. AB - Digital fingerprinting is a technology for tracing the distribution of multimedia content and protecting them from unauthorized redistribution. Unique identification information is embedded into each distributed copy of multimedia signal and serves as a digital fingerprint. Collusion attack is a cost-effective attack against digital fingerprinting, where colluders combine several copies with the same content but different fingerprints to remove or attenuate the original fingerprints. In this paper, we investigate the average collusion attack and several basic nonlinear collusions on independent Gaussian fingerprints, and study their effectiveness and the impact on the perceptual quality. With unbounded Gaussian fingerprints, perceivable distortion may exist in the fingerprinted copies as well as the copies after the collusion attacks. In order to remove this perceptual distortion, we introduce bounded Gaussian-like fingerprints and study their performance under collusion attacks. We also study several commonly used detection statistics and analyze their performance under collusion attacks. We further propose a preprocessing technique of the extracted fingerprints specifically for collusion scenarios to improve the detection performance. PMID- 15887560 TI - Video content classification based on 3-D eigen analysis. AB - To achieve video understanding, it is of utmost practical importance to classify videos according to its spatial and temporal features in an efficient and effective manner. It still remains, however, largely an elusive task. In still image analysis, thanks to the great efforts made by many researchers, a broad spectrum of methods have been developed with great success, especially the ones based on eigen analysis due to its efficacy. In this paper, inspired by the impressive performance achieved by this framework, we will develop a content based video classification method based on three-dimensional (3-D) eigen analysis. Unlike most other video understanding schemes where the spatial and temporal contents play different roles in the processing, this new method treats a video as a solid within a 3-D Euclidean space and can, thus, naturally take advantage of the spatial and temporal contents existing in videos. After computing the eigen values and corresponding eigen vectors of the autocorrelation matrix for each small 3-D macroblock, different labels are assigned regarding its spatial/temporal natures based on the behavioral properties of the eigen values and eigen vectors. Extensive empirical studies have suggested encouraging performance for the use of this eigen analysis-based video classification method. PMID- 15887561 TI - Inkjet printer model-based halftoning. AB - The quality of halftone prints produced by inkjet (IJ) printers can be limited by random dot-placement errors. While a large literature addresses model-based halftoning for electrophotographic printers, little work has been done on model based halftoning for IJ printers. In this paper, we propose model-based approaches to both iterative least-squares halftoning and tone-dependent error diffusion (TDED). The particular approach to iterative least-squares halftoning that we use is direct binary search (DBS). For DBS, we use a stochastic model for the equivalent gray-scale image, based on measured dot statistics of printed IJ halftone patterns. For TDED, we train the tone-dependent weights and thresholds to mimic the spectrum of halftone textures generated by model-based DBS. We do this under a metric that enforces both the correct radially averaged spectral profile and angular symmetry at each radial frequency. Experimental results generated with simulated printers and a real printer show that both IJ model based DBS and IJ model-based TDED very effectively suppress IJ printer-induced artifacts. PMID- 15887562 TI - A computational model for color naming and describing color composition of images. AB - The extraction of high-level color descriptors is an increasingly important problem, as these descriptions often provide links to image content. When combined with image segmentation, color naming can be used to select objects by color, describe the appearance of the image, and generate semantic annotations. This paper presents a computational model for color categorization and naming and extraction of color composition. In this paper, we start from the National Bureau of Standards' recommendation for color names, and through subjective experiments, we develop our color vocabulary and syntax. To assign a color name from the vocabulary to an arbitrary input color, we then design a perceptually based color naming metric. The proposed algorithm follows relevant neurophysiological findings and studies on human color categorization. Finally, we extend the algorithm and develop a scheme for extracting the color composition of a complex image. According to our results, the proposed method identifies known color regions in different color spaces accurately, the color names assigned to randomly selected colors agree with human judgments, and the description of the color composition of complex scenes is consistent with human observations. PMID- 15887563 TI - Development of a method for the concentration and recovery of microsporidia from tap water. AB - Microsporidia are obligate intracellular parasites. Microsporidian spores infect a wide variety of hosts, including humans. The spores may be found in infected hosts' urine and feces, thus waterborne transmission is possible. This study details method development for the detection of microsporidia in tap water. In this study, filtration, centrifugation, purification, and detection parameters were optimized for the detection of microsporidia. The Pall-Gelman Envirocheck sampling capsule (Pall Gelman, Ann Arbor, MI) was chosen as the filter element. Optimal centrifugal force for spore recovery was 1500 x g. Additionally, it was determined that eluting microsporidia spores in a detergent elution buffer solution had a detrimental effect on spore recovery. A direct examination of the concentrate resulted in a greater recovery with less variability than subjecting the sample concentrate to a Percoll-sucrose density gradient purification step. The staining method employed for the detection spores was Calcofluor white (Sigma, St. Louis, MO). Percent recoveries for 10 L tap water samples (n = 5) using the Envirocheck sampling capsule without a density gradient purification step were 26.1+/-13.4 compared to 25+/-13.8 for samples subjected to a density gradient purification step. PMID- 15887564 TI - Differential effects of fluoranthene and benzo[a]pyrene in MCF-7 cells. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants, which are suspected carcinogens and may affect the reproductive system as potential endocrine disruptors. Therefore, we tested fluoranthene (FL) and benzo[a]pyrene (BaP) on human breast cancer cell line (MCF-7 cells) to determine possible toxic effects. The cells were incubated in the presence of medium, medium containing 0.1% dimethylsulfoxide (DMSO) as vehicle, or in the presence of FL (10, 50, and 100 microg/ml), BaP (10, 50, and 100 microg/ml), 17beta-estradiol (E2; 5 microg/ml and 500 ng/ml), or tamoxifen (Tx; 5 microg/ml and 500 ng/ml). After 24 h, FL (100 microg/ml), BaP (100 microg/ml), or Tx (5 microg/ml) killed significant numbers of cells. After 72 h, FL (50 and 100 microg/ml), BaP (100 microg/ml), E2 (5 microg/ml), or Tx (5 microg/ml and 500 ng/ml) decreased MCF-7 cell viability significantly as demonstrated by the MTT assay. Measurement of DNA synthesis was conducted using 3H-thymidine incorporation into MCF-7 cell DNA for 72 h. After 72 h, BaP (10, 50, and 100 microg/ml) and Tx (5 microg/ml and 500 ng/ml) significantly decreased DNA synthesis in MCF-7 cells. FL did not significantly alter 3H-thymidine incorporation into the cells. While higher concentration of E2 (5 microg/ml) decreased 3H-thymidine incorporation, the lower concentration of E2 (500 ng/ml) increased cell proliferation. Apoptotic response was tested by in situ fluorescence staining of cells incubated for 72 h in media containing 0.1% DMSO, or vehicle containing FL (10 microg/ml), BaP (10 microg/ml), E2 (500 ng/ml), or Tx (500 ng/ml). Microscopic examination demonstrated presence of apoptosis with BaP (10 microg/ml) and Tx (500 ng/ml), but not with FL (10 microg/ml) and E2 (500 ng/ml). The cell cycle analysis using flow cytometry demonstrated that E2 (500 ng/ml) did not significantly change the progression of MCF-7 cells after 72 h of incubation. However, FL (10 microg/ml) only suppressed G2/M phase. Tx (500 ng/ml) blocked G0/G1, S, and G2/M phases, and BaP (10 microg/ml) suppressed the G0/G1 phase. These data suggest that BaP on MCF-7 cells is growth inhibitory and apoptotic, whereas the toxic effects of FL are not exerted through apoptosis. PMID- 15887565 TI - Comparative analysis of nitrifying bacteria in full-scale oxidation ditch and aerated nitrification biofilter by using fluorescent in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE). AB - In this study, nitrification performances and composition of nitrifying populations in a full-scale oxidation ditch and a high-rate submerged media nitrification biofilter were comparatively analyzed. In addition to different reactor configurations, effects of differing operational conditions on the nitrification efficiency and bacterial diversity were also explored and evaluated thoroughly. In microbial analysis of sludge samples fluorescent in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) techniques were used complementary to each other. The extended aeration oxidation ditch subjected to the study is operated as a nitrogen and phosphorus removal system consisting of anaerobic, anoxic, and aerobic zones. The high-rate submerged media aerated filter is operated as nitrification step following the conventional activated sludge unit and the nitrified wastewater is discharged to the sea without complete nitrogen removal. In situ hybridization results have indicated that Nitrosomonas-like ammonia oxidizing and Nitrospira-related nitrite oxidizing bacteria were intensively present in vigorous flocs in nitrification biofilter while carbonaceous bacteria belong to beta subclass of Proteobacteria were considerably dominant in oxidation ditch. Low quantities of nitrifiers in oxidation ditch were also confirmed by the dissimilarity in intensive bands between two systems obtained with DGGE analysis. PMID- 15887566 TI - Cytotoxicity and mutagenicity study of waste and purified water samples from electroplating industries prepared by use of ferrous sulfate and wood fly ash. AB - Toxicological safety of the new purification method for electroplating wastewaters (EWW) has been assessed. Method utilizes waste by-product ferrous sulfate and wood fly ash to scavenge heavy metals from EWW. Energy dispersive X ray fluorescence (EDXRF) analysis of all samples met the law requirements for safe depositing of waste into the environment. The toxicity study consisted of determination of frequency of bacterial and human cell lines survival and the Ames assay (plate incorporation assay and the preincubation assay). Unexpectedly, data obtained indicate cytotoxicity and slight mutagenic potential of purified water. Ames assay showed that combination of alkalis and heavy metals present in the water purified with original fly ash and eluates of fly and waste ash were metabolically activated and conjugated with glutathione, resulting in new metabolites active and toxic for the cell. In order to reduce this effect pretreatment of fly ash (partial removal of highly soluble compounds) are necessary prior to its usage for neutralization /coagulation/flocculation processes. PMID- 15887567 TI - Temporal and spatial assessment of water quality, physical habitat, and benthic communities in an impaired agricultural stream in California's San Joaquin Valley. AB - The goal of this study was to characterize and discuss the relationships among water quality, physical habitat, and benthic community data collected annually over a three-year period (2000--2002) in an impaired agricultural stream (Orestimba Creek) in California's San Joaquin River watershed. Conductivity, pH, and turbidity were the most important water quality conditions influencing the various benthic metrics. Significantly higher flow conditions and lower dissolved oxygen values were reported in Orestimba Creek in 2001; increased turbidity conditions were reported in 2002. Channel alteration, riparian buffer, sediment deposition, and channel flow were the most important physical habitat metrics influencing the various benthic metrics. Higher total physical habitat scores were reported in 2001 when compared with 2002. The most dominant benthic taxa collected during all three years of sampling were oligochaetes and chironomids. Oligochaetes are found in stressful environments while chironomids can be either sensitive or tolerant to environmental stressors depending on the species. Populations of both daphnids and the exotic clam Corbicula were reported to increase over time. Both of these taxa are generally tolerant to most types of environmental degradation. The exception is that daphnids are highly sensitive to organophosphate insecticides. The % filterers increased over time, which suggests an increase in environmental disturbance. The % collectors decreased from 2000 to 2002, which suggests an improvement in environmental conditions. The presence of approximately 100 taxa in Orestimba Creek during each of the three years of sampling implies that benthic communities in this stream are fairly diverse, considering their ephemeral environment, but without a clear definition of benthic community expectations based on established referenc conditions it is unknown if this water body is actually impaired. PMID- 15887568 TI - A-role of alkaline elements in formation of PCDDs, PCDFs, and coplanar PCBs during combustion of various paper samples. AB - Combustion of industrial and domestic wastes produces significant amounts of dioxins (PCDDs, PCDFs, and coplanar PCBs). These wastes contain numerous kinds of materials, including inorganic elements, which may play an important role in dioxin formation upon combustion. Six paper samples--impregnated with NaHCO3, NaCl + fly ash, polyvinyl chloride (PVC), and PVC + NaHCO3--were combusted in a well-controlled incinerator at 512-760 degrees C and exhaust gases were analyzed for dioxins (PCDDs, PCDFs, and coplanar PCBs). Addition of fly ash, which contained high levels of Ca and Cl, to NaCl impregnated-newspaper increased total dioxin formation from 80.8 ngg(-1) to 139 ngg(-1) during combustion. Addition of NaHCO3 to PVC-impregnated newspaper increased total dioxin formation from 220 ngg(-1) to 288 ngg(-1). Formation of PCDDs from NaHCO3/PVC-impregnated newspaper were significantly less than that from newspaper impregnated with PVC alone. On the other hand, more PCDFs formed from NaHCO3/PVC-impregnated newspaper than from newspaper impregnated with PVC alone. Addition of NaHCO3 to a PVC-impregnated newspaper changed the production pattern of dioxins. PVC- impregnated newspaper produced the greatest amount of coplanar PCBs (12.3 ngg(-1)), which may be accounted for by the relatively high level of TEQ value (3.31 ng-TEQg(-1)) of this sample. Generally, addition of fly ash and alkali metals, such as Na, to paper samples increased the total dioxin formation in exhaust gases during combustion. The results suggest that alkali metals increase Cl concentrations at a reaction site by trapping chloride ions and/or chloride radicals and consequently increasing dioxin formation. PMID- 15887569 TI - Photocatalytic treatment of linear alkylbenzene sulfonate (LAS) in water. AB - The photocatalytic degradation of aqueous linear alkylbenzene sulfonate (LAS) was studied. Two different photocatalysts, Degussa P25 TiO2 and Hombikat UV 100 TiO2, were used to degrade aqueous linear alkylbenzene sulfonate in slurry batch photoreactors. For a 100 mg/L LAS solution based on first-order rate constants, the optimum photocatalyst loading for Degussa P25 TiO2 was 4.0 g/L, while for Hombikat UV 100 TiO2 it was 2.0 g/L. The photoactivity of Degussa P25 TiO2 it was higher than that of Hombikat UV 100 TiO2 for the treatment of LAS. A mixture of both photocatalysts did not improve the LAS degradation rates in batch experiments. Combination of Degussa P25 TiO2 and 600 mg/L H2O2 along with irradiation with UV light at either 254 or 365 nm did not improve the LAS degradation rates over the photocatalytic or photolytic processes individually. PMID- 15887570 TI - Effect of additives on photodegradation of 1,3,5-trichlorobenzene in aqueous surfactant solutions. AB - Photolysis of 1,3,5-trichlorobenzene (TCB) in moderate concentration (0.5 mM) in the presence of several additives was examined in 10 mM of cationic and nonionic surfactant solutions. Additions of small amounts of hydrophobic additives, n dodecanethiol (1 mM), n-dodecyldimethylamine (<2.5 mM), and N-(n-dodecyl)-N methylaniline (C12An: <0.5 mM), were effective for photodechlorination of TCB, while the formation of by-products could not be inhibited perfectly. In contrast, exclusive and efficient dechlorination of TCB in cetyltrimethylammonium chloride solution was achieved in the presence of sodium borohydride (<5 mM), which was due to the enhanced local concentration of borohydride anions in cationic micelle surfaces. PMID- 15887571 TI - Wastewater treatment using fibrist and saprist peat: a comparative study. AB - Two different peat types (fibrist and saprist) were compared to examine their respective wastewater treatment capacity in peat beds under different hydraulic loadings (1.8, 1.2, 0.9, and 0.6 m3/m2 d) and pollutant loadings (dependent on influent quality). The comparative study was carried out on a pilot scale using urban wastewater which had undergone preliminary treatment. The systems were observed to retain a higher quantity of suspended solids when the load was higher, regardless of the hydraulic loading and the type of peat employed. System performance with respect to BOD was similar to that observed for COD, and it was noted that the capacity to retain organic matter decreased when the hydraulic loading or pollutant loading was increased. This effect was observed in both peats but to different degrees, being lower in the case of saprist peat. The results indicate that it would be necessary to work with low hydraulic loadings in order to comply with legislative requirements on effluent. Peat beds are therefore suitable for implementation in small population centers, where the system's efficiency partly depends on the type of peat employed. In view of the system's performance with respect to suspended solids, this technology may be considered a good primary treatment, and could be usefully combined with other processes aimed at eliminating dissolved organic matter. PMID- 15887572 TI - Oxidative degradation and toxicity reduction of trichloroethylene (TCE) in water using TiO2/solar light: comparative study of TiO2 slurry and immobilized systems. AB - A solar-driven, photocatalyzed degradation system using TiO2 slurry and immobilized systems was constructed and applied to the degradation of trichloroethylene (TCE) contaminated water using TiO2 with solar light. The experiments were carried out under constant weather conditions on a sunny day. Solar photocatalytic treatment efficiency of the solar light/TiO2 slurry system was compared with that of the solar light/TiO2 immobilized system. The operation of the solar light/TiO2 slurry and immobilized systems showed 100% (TiO2 slurry system), 80% (TiO2 immobilized system) degradation of the TCE after 6 h, with a chloride production yield of approximately 89% (TiO2 slurry system), 72% (TiO2 immobilized system). The oxidants such as H2O2 and S2O8(2-) in the TiO2 slurry and immobilized systems increased TCE degradation rate by suppressing the electron/hole recombination process. The degradation rate and relative toxicity reduction of TCE followed the order of solar light/TiO2 slurry + S2O8(2-) > solar light/TiO2 slurry + H2O2 > solar light/TiO2 immobilized + S2O8(2-) > solar light/TiO2 slurry > solar light/TiO2 immobilized + H2O2 > solar light/TiO2 immobilized. Finally, following to the toxicity result, the acute toxicity was reduced by below toxicity endpoint (EC50 concentration) following the treatment. It means that many of the metabolites of TCE reduction are less toxic to Vibrio fischeri than the parent compound. Based on these results, TCE can be efficiently and safely treated in a solar-driven, photocatalyzed degradation system. PMID- 15887573 TI - Separation of Tl(I) and Tl(III) from environmental water samples by flotation method coupled with Zeeman ETAAS determination. AB - An accurate, fast, and simple method for determination of thallium in environmental water samples with Zeeman electrothermal atomic absorption spectrometry (ZETAAS) by previous flotation preconcentration is given. The possibility of using cobalt(III) hexamethylenedithiocarbamate as a colloidal precipitate collector for simultaneous flotation separation of Tl(I) and Tl(III) from water matrices was studied. All experimental parameters influencing a proper flotation of both thallium ions were ascertained. The developed procedure with Co(HMDTC)3 is applied for preconcentration and separation of total thallium in water matrix before ZETAAS. The results of ZETAAS analysis are compared with those obtained by inductively coupled plasma-atomic emission spectrometry. The ZETAAS limit of detection of thallium preconcentrated by Co(HMDTC)3 is 0.031 microg/L. PMID- 15887574 TI - Zirconia pillared montmorillonite for removal of arsenate from water. AB - Zirconia pillared montmorillonite, a clean adsorbent with increased specific areas of 40.118 m2/g and high basal spacing of 2.20 nm, was prepared for the removal of arsenate from water. Zirconia pillared montmorillonite is effective for the removal of arsenate. Adsorption is favored under acid conditions. Ca2+ and Mg2+ in the solution slightly enhance the adsorption. Over 95% removal was observed under natural pH conditions from 20 mg/L arsenate solutions containing 5.0 mg/L Ca2+ or Mg2+. High adsorption capacity of over 120 mg/g was observed from arsenate solutions with different Ca2+ concentrations. The Freundlich model can describe the adsorption equilibrium data well. The column test shows that the column allows the passage of the feeding solution for approximately 225 times the bed volume if the removal percentage of arsenate was monitored not to be lower than 98%. The adsorbed arsenate can be desorbed with NaOH and the desorption efficiency reaches 85% and 88% when the concentration of NaOH reaches 0.2 and 0.5 mol/L. Zirconia pillared montmorillonite can be regenerated and the regenerated adsorbents still have good adsorption capacities. PMID- 15887575 TI - Orthogonal signal correction used for noise elimination of open path Fourier transform infrared spectra. AB - Open path Fourier transform infrared (OP-FTIR) spectroscopy is qualified in detecting mixtures by multivariate calibration methods such as partial least squares (PLS); however, its applications are still restricted by background noise, which is unavoidable for OP-FTIR spectra and cannot be resolved solely by multivariate calibration methods. Hence OP-FTIR spectra are often pretreated before the data are subjected to the multivariate calibration model. A new preprocessing technique, orthogonal signal correction (OSC), was presented in this paper. The principle of OSC is to remove the part in X orthogonal to Y, and it is implemented based on PLS and nonlinear iterative partial least squares (NIPALS) algorithm. The approach was applied to three different data sets of PLS model. It performed much better than classical PLS when handling data with noise but comparably when processing on the simulated data. Moreover, OSC could reduce the complexity of model, which would facilitate the interpretation of the models. The results reveal that the proposed method gives much better prediction than the classical PLS and is very promising for the wide use of OP-FTIR. The preprocessing technique, auto-scaling, and second-order derivatives (SOD) were also considered. PMID- 15887576 TI - Root responses and metal accumulation in two contrasting ecotypes of Sedum alfredii Hance under lead and zinc toxic stress. AB - Sedum alfredii Hance has been reported to be a Zn-hyperaccumulator plant species. In this study, root morphological and physiological response of the hyperaccumulating ecotype of S. alfredii H. (HE) from the mined area and the non hyperaccumulating ecotype of S. alfredii H. (NHE) from the agricultural area to supplied levels of Zn and Pb were investigated. The results showed that Zn concentrations in the leaves and the stems of the HE were 34 and 41 times higher, whereas lead concentrations were 1.9 and 2.4 times greater, respectively, than those of the NHE when grown at 1224 microM Zn and/or 200 microM Pb. At combined supply of 1224 microM Zn with 200 microM Pb, however, zinc concentrations in the stems and leaves of the, HE decreased, while lead concentrations in the stems increased significantly, as compared with those of single metal treatment. Lead uptake of the HE was enhanced by Zn addition. Root activity of the HE decreased by Pb treatment in the first two days, but recovered afterward and close to the control at day 10 of the treatment. However, root activity of the NHE decreased by each metal treatment, and was not recovered with the advance of treatment time. Root length, root surface area, and root volumes increased obviously due to Zn and/or Pb/Zn combined treatments for the HE, but significantly decreased due to Pb, Zn, or Pb/Zn combined treatment for the NHE. Zinc and Pb concentrations in both ecotypes of S. alfredii H. were positively correlated with root length, root surface area, and root volumes. Root exudates of the HE, especially treated with Zn, increased the extractability of Pb and Zn from the mined soil. At the Zn supply level of 1224microM, the extractability of root exudates on soil Pb was 3 12 times greater for the HE than for the NHE. These results imply that the tolerance and hyperaccumulation of the hyperaccumulating ecotype of S. alfredii H. to Zn and Pb appear to be closely related to its high adaptation of root growth, morphology, and physiology to Pb and Zn toxicity, and through its root excretion of some special substances that can activate Pb and Zn in the mined soil, thus increasing their mobilization and bioavailability. PMID- 15887577 TI - Seminal concentration of trace elements in fox and relationships to spermatozoa quality. AB - Concentrations of copper, zinc, iron, cadmium, lead, and nickel in the semen of foxes (Vulpes vulpes, n = 10), microscopic analysis of occurrence of pathological spermatozoa, and correlations of these elements with pathological forms were studied. Samples were analyzed by using an atomic absorption spectrophotometer. For analysis of pathological spermatozoa semen samples fixed with Hancock's solution and stained with Giemsa were prepared. For each fox at least 1000 spermatozoa were evaluated. The concentrations of copper, zinc, and iron in semen of foxes were found to be 2.16+/-0.53 mg/kg, 13.09+/-5.22 mg/kg, and 33.16+/ 24.36 mg/kg, respectively, on wet weight basis. Concentration of cadmium was low (0.07+/-0.05 mg/kg). The levels of lead and nickel in the semen of foxes were 0.08+/-0.06 mg/kg and 0.35+/-0.24 mg/kg, respectively. The total percentage of pathological spermatozoa was 7.76+/-1.33% with predominancy of knob twisted flagellum, separated flagellum, and broken flagellum. In relation to trace elements the analysis showed significant (p < 0.05) correlation between copper and lead (r = -0.85), copper and other forms of pathological spermatozoa (r = 0.72), zinc and broken flagellum (r = -0.69), iron and retention of cytoplasmic drop (r = 0.87), cadmium and separated flagellum (r = -0.68), and between cadmium retention of cytoplasmic drop (r = 0.87). PMID- 15887578 TI - Radiologic case study. First metarsophalangeal plantar plate injury (turf toe). AB - Turf toe and related injuries to the first metatarsophalangeal joint are common injuries that occur in several sports. Although often a clinical diagnosis, advanced imaging can help grade severity of sprain and evaluate for associated or unsuspected injuries. Without proper rest and conservative management, a treatable injury can have chronic sequelae and morbidity. Operative management is uncommon, but successful in returning high-level competitive athletes to their sport. PMID- 15887579 TI - Practice management. PMID- 15887580 TI - Extending the width of an operating table using armboards. PMID- 15887581 TI - Cincinnati incision combined with medial rotation fasciocutaneous flap for clubfeet with pathologic soft tissues. PMID- 15887582 TI - Don't get burned: beware of drug-induced photosensitivity reactions! PMID- 15887583 TI - Duthie's biological repair of ruptured Achilles tendons. PMID- 15887584 TI - Evidence-based approaches to minimizing malpractice risk in orthopedic surgery. AB - The malpractice crisis continues unabated in the United States. However, there are many strategies with foundations in scientific study that physicians can use to reduce their malpractice risk. Studies have shown technical error has little relationship with malpractice claims. Patients with higher education levels and socioeconomic status are more likely to file lawsuits. Surgeons should be aware of how to respond in the event a claim is filed. Improving physician-patient communication is the cornerstone of any strategy to reduce malpractice claims. PMID- 15887585 TI - Management of irreparable rotator cuff tears and glenohumeral arthritis. AB - Glenohumeral arthritis with irreparable rotator cuff tears remain a difficult entity to treat. Varied causes include rotator cuff tear arthropathy, osteoarthritis, or rheumatoid arthritis with irreparable cuff tear. Common symptoms are progressive pain and dysfunction. Physical examination may reveal pain, crepitance, rotator cuff weakness, and loss of motion and function. Radiographs may reveal varying degrees of osteophyte formation, sclerotic bone, superior humeral head migration, and bony erosion. Additional imaging modalities may reveal cuff tear size, retraction, atrophy, and fatty infiltration. Failure of nonoperative management may lead to operative intervention. Rotator cuff repair or reconstruction may help prevent progression of tears and future arthritic changes. In patients with moderate to severe glenohumeral arthritis and irreparable rotator cuff tears, hemiarthroplasty is currently the procedure of choice. For patients with severe cuff dysfunction or loss of coracoacromial arch, or for patients who require revision, the reverse shoulder prosthesis may offer a treatment option. Future management continues to be defined with additional study. PMID- 15887586 TI - The effect of radiofrequency energy on nonweight-bearing areas of bone following shoulder and knee arthroscopy. AB - This prospective randomized clinical trial evaluated whether the use of radiofrequency energy (RFE) devices for soft-tissue ablation and coagulation cause thermal injury to bone. Fifty patients underwent one of three treatment modalities: electrocautery, monopolar RFE, or bipolar RFE. Preoperative and postoperative magnetic resonance imaging was compared to evaluate for evidence of osteonecrosis. Postoperative MRI of all patients did not reveal any osteonecrosis or subchondral edema. These findings indicate electrocautery, monopolar RFE, and bipolar RFE devices can be used safely for soft-tissue blation and hemostasis. PMID- 15887587 TI - The effect of drill-hole length on the primary stability of osteochondral grafts in mosaicplasty. AB - Osteochondral grafts were transplanted from the trochlea of porcine femurs into drill holes that were 20, 15, and 12 mm in length on the lateral femoral condyle. Grafts initially were pushed in flush with the surrounding cartilage, and then a testing machine pushed the grafts 3 mm deeper. For the 20-, 15-, and 12-mm drill holes, mean forces for pushing the graft flush were 36.58, 43.33, and 118.13 N, respectively, while mean forces for pushing the graft 3 mm deeper were 122.50, 249.33, and 377.25 N, respectively. These results suggest primary stability is better when grafts and drill holes are the same length, but if the recipient hole is shorter, excessive force must be exerted on the cartilage cup during insertion. PMID- 15887588 TI - Dysfunction of the pisotriquetral joint: degenerative arthritis treated by excision of the pisiform. AB - From 1995 to 2000, 21 patients (14 women and 7 men) with a mean age of 42 were treated with excision of the pisiform for a dysfunction of the pisotriquetral joint. Follow-up ranged from 6 to 36 months (average: 30 months). The diagnoses included degenerative arthritis of the pisotriquetral joint (15 patients), degenerative arthritis associated with a ganglion (3 patients), and calcifications caused by flexor carpi ulnaris tendinopathy (3 patients). All patients had pain secondary to direct pressure on the pisiform. Side-to-side passive motion of the pisiform occasionally led to pain and crepitus. Degenerative arthritis and calcifications in the pisotriquetral joint were confirmed by a wrist radiograph (lateral view in 30 degrees supination). In five patients, local injection with anesthetic temporarily resolved the symptoms. Excision of the pisiform resulted in complete relief of pain without functional deficit. PMID- 15887589 TI - Ipsilateral stress fracture of the proximal tibia after primary hybrid total hip arthroplasty. PMID- 15887590 TI - Systemic lead poisoning due to an intra-articular bullet. PMID- 15887591 TI - Negative-pressure pulmonary edema in the arthroscopic patient. PMID- 15887592 TI - Substance use among young adolescents in HIV-affected families: resiliency, peer deviance, and family functioning. AB - This study examines the association of risk and protective factors with substance use among 77 early adolescents (11-15 years old) with an HIV-infected parent who were interviewed in 2000-2001 in the South Bronx, a HIV high-prevalence area of New York City. The subjects were 49%female, 53% African American, and 30% Hispanic; mean age was 13 years old. A face-to-face interview was used to administer a battery of instruments representing community, family, peer, and resiliency factors. Forty percent reported ever using tobacco, alcohol or drugs; 71% were aware of their parent's HIV seropositivity. An age-adjusted path analytic model was constructed which showed: 1) family functioning predicted resiliency (a composite measure of psychological adjustment and personal competencies); 2)positive community factors and resiliency predicted less affiliation with deviant peers; and 3) poorer family functioning and affiliation with deviant peers predicted substance use. These results underscore the need for interventions that address social influence factors among vulnerable early adolescents with HIV-positive parents. PMID- 15887593 TI - Assessing attachment cognitions and their associations with depression in youth with eating or drug misuse disorders. AB - The study investigates associations between attachment cognitions and depression symptoms in 71 15-25-year-olds, 26 of whom have eating disorders, and 20 of whom are drug misusers. Attachment cognitions were measured with the CaMir Q-sort, which provides indexes for secure, avoidant, and preoccupied attachment, as well as scores on 13 dimensions. The BDI-13 was used to measure depressive symptomatology. Consistent with the literature, BDI scores were associated with cognitions of preoccupied attachment. They were also related to cognitions of avoidant attachment, confirming Bowlby's theory on defensive exclusion. For participants with eating disorders, depressive symptomatology was related to preoccupation and parental interference, whereas for drug misusers, it was negatively related to security, preoccupation, parental support, and lack of parental concern. These findings help understand how attachment cognitions may participate in depressive symptomatology, namely in youth whose behavior problems may be associated with specific attachment experiences. PMID- 15887594 TI - Attitudes of teenagers towards cigarettes and smoking initiation. AB - This paper contributes to the empirical knowledge of determinants of smoking initiation in adolescents. The instrument we used was a structural equation model, which is a powerful tool to analyze causal relationships in nonexperimental studies. We used a school-based sample of 1198 teenagers from Spain. We measured the attitude of the adolescents towards tobacco. Attitudes and smoking status are related. Attitudes become more favorable to smoking, as experimentation with cigarettes progresses and pupils become older. Teenagers who have tried at least one cigarette are on average more favorable to smoking than those who had never smoked. The study's limitations were noted. PMID- 15887595 TI - Changes in reasons given for adolescent smoking, 1984-1999. AB - Adolescent smoking has been widely studied, but surprisingly little research has been done about the reasons given for smoking by adolescents themselves. This study examines the reasons given by Finnish adolescents for their own smoking and the reasons that they perceive for smoking by others. It reports on how these reasons have changed over a period of 15 years. In 1984, a questionnaire about reasons for smoking was administered to a sample of adolescents aged 14-16 (N = 396). The questionnaire was administered again to a similar sample (N = 488) in 1999, when Finland adopted strict new tobacco legislation. It was found that the reasons given (i.e., attributions) had changed considerably, and that the attributions for the adolescents' own behavior were quite different from the attributions for smoking by others. The attributions were only weakly influenced by the participants' gender or by their smoking habits, either in 1984 or 1999. In relation to participants' own smoking, the later questionnaire elicited inner subjective experiences involving "good feelings." In relation to the perceived reasons for other people's smoking, it elicited more responses connected with the notion of "belonging." The limitations of the study and suggestions for further research are discussed. PMID- 15887596 TI - Reliability and concurrent validity of the CAGE screening questions: A comparison of lesbians and heterosexual women. AB - Although lesbians are believed to be at heightened risk for alcohol abuse and alcohol dependency, the reliability and validity of currently available screening measures have yet to be explored in this population group. In this paper, we report the psychometric properties of the CAGE in a diverse sample of 63 lesbians and comparisons with a control group of 57 heterosexual women. Data were collected in Chicago during 1997-1998 using face-to-face interviews. Findings suggest that the CAGE has good reliability and concurrent validity among lesbians. One CAGE item, concerned with drinking in the morning, however, was found to be only weakly associated with the other items among lesbians. Although this pilot study is limited by the nonprobability sample, we conclude that evaluating the appropriateness of other alcohol and drug abuse measures in high risk populations, such as lesbians, should be a research priority. PMID- 15887597 TI - An examination of drug activity: effects of neighborhood social organization on the development of drug distribution systems. AB - OBJECTIVE: The ability to determine the geographic locations of illicit drug markets is central to the development of preventive interventions that address access to drugs and associated problems, such as violence and crime. METHOD: The current study examined individual self-reports of drug activities and demographic information obtained from two waves of a telephone survey of 1704 individuals aged 15 to 29 conducted in 1999 and 2001 across 95 census tracts in a Northern California city and measures of neighborhood characteristics derived from Census 2000 measures. RESULTS: The results of the study showed that, at the individual level, younger people and male respondents reported most drug activities. At the aggregate level, neighborhood poverty was directly related to higher rates of drug activity. Residential stability was found to moderate reports of drug activity observed by African-Americans and young people. CONCLUSION: Social processes reflected in neighborhood characteristics of census tracts influence rates of self-reports of individuals' exposures to drug activities. PMID- 15887598 TI - Smoking and drinking patterns among residents of Li ethnic minority villages in Hainan, China. AB - BACKGROUND: In China, approximately 67% of male adults are smokers and 64% of them are drinkers. Tobacco and alcohol use are more prevalent among many of China's 55 ethnic minority populations, but specific data are lacking for them, including the Li population. METHODS: Cross-sectional data from 379 residents sampled from 12 Li villages in Hainan Province were used for this analysis. Data were collected in 2003 using a questionnaire adopted from the U.S. National Household Survey on Drug Abuse. Both lifetime and current use of tobacco and alcohol products, including prevalence, frequency and amount of use, as well as age patterns of onset of use were assessed. FINDINGS: Eighty-five percent of the sample were Li minority and 43% were female with a mean age of 34.7 years (SD = 14.5). While fewer female subjects of the sample reported smoking (1.9% of lifetime smoking) and drinking (21.0% lifetime drinking), 84.1% of the male sample were lifetime smokers and 92.5% were lifetime drinkers; 78.1% were 30-day smokers and 88.4% were 30-day drinkers; and 58.6% were daily smokers and 15.8% were daily drinkers. In addition, 79.1% male villagers were binge drinkers, and 21.4% were intoxicated in the past month. The risk of onset of smoking and drinking increased after 10 years of age, peaked at 18-19 years (hazard = 16.0% for smoking and 19.5% for drinking), then declined until leveling off at 25 years of age. Eighty-five to 97% of the males simultaneously used alcohol and tobacco. CONCLUSIONS: The prevalence rates of tobacco and alcohol use in Li male villagers are higher than those of the national average and other ethnic minorities in China. Indepth studies are needed to obtain more detailed information regarding issues such as detailed types of tobacco and alcohol products, causes of use, as well as willingness to participate in tobacco and alcohol control interventions in order to develop and deliver substantial intervention programs. PMID- 15887599 TI - Sexual behavior patterns of methamphetamine-using gay and bisexual men. AB - We assessed the sexual behaviors of 49 gay and bisexual methamphetamine users in New York City and the relationship between their use of the substance and their sexual practices as part of a larger investigation of the behavioral aspects of methamphetamine use. Participants were assessed on their use of methamphetamine and their sexual behaviors in conjunction with use of the substance. Participants reported equivalent rates of unprotected anal insertive and receptive behaviors when comparing their sexual acts while high on methamphetamine, high on other drugs, and sober. In addition, equivalent rates of "extreme" sex acts were found for 10 of the 12 behaviors examined while high on methamphetamine and while sober. While wefound few differences in terms of rates of sexual behaviors, our analysis revealed more frequent risky sexual behaviors among HIV positive men when compared with HIV negative men. Our results suggest that methamphetamine attracts a hypersexual risk-taking group of men who engage in unprotected sexual behaviors regardless of their methamphetamine use. Treatment implications are discussed. PMID- 15887600 TI - Juvenile drug use and delinquency: youths' accounts of their trajectories. AB - This study focuses mainly on youths' opinions about their delinquency and drug use trajectories. A qualitative approach was taken for this phenomenological research. The life account method was used to conduct a total of 62 interviews: 36 boys and 26 girls recruited in youth detention and addiction treatment centers, youth centers, high schools, and on the streets of three cities in Quebec between 1996 and 2002. Thematic analysis was used, and the deviant trajectories of the youths are broken down into five stages. Our study provides an incomplete view of these youths' more or less deviant trajectories. It would be interesting to do a follow-up study. PMID- 15887601 TI - Bio-electric approaches for youths; treating post-stroke depression. PMID- 15887602 TI - A story of transformation: an agency fully embraces recovery. PMID- 15887603 TI - Managing to hold down Medicaid drug costs. PMID- 15887604 TI - A smoother transition from inpatient to outpatient. PMID- 15887605 TI - Automating financial and clinical operations: tough questions and good advice. PMID- 15887606 TI - Whackamole? The role of key performance indicators in health and human services. PMID- 15887607 TI - Crisis response for business: more than an intervention. PMID- 15887608 TI - Another nation's approach to system-building. PMID- 15887609 TI - Physicians' liability in obstetric and gynecology practice. AB - PURPOSE: Litigation in obstetric and gynecologic practice has arisen as an important issue. This article aims to highlight issues related to malpractice in obstetrics and gynecology in order to point out several important aspects of medical practice, which may offer adequate professional protection. METHODS AND MATERIAL: Search of the current relevant literature. RESULTS: The four most common clinical causes of medical litigation in obstetrics and gynecology are cases involving fetal distress, uterine rupture after a vaginal birth in a woman with a previous Cesarean section, shoulder dystocia, and misdiagnosis of breast cancer. Litigation raised in most of the cases is driven by bad outcomes and not by malpractice. Additionally, both juries and the public often expect perfection as the natural result of pregnancy and any deviation from this expectation has to be the result of someone's negligence. For this reason it has become mandatory for doctors in hospitals and private practices to acquire a sufficient knowledge relating to modem management, to document carefully in writing their procedural and management plans and to become familiar with the legal aspects of medical practice. Adequate communication and a written patient's consent form are of paramount importance. CONCLUSION: Maintenance of high standards in daily practice with continuous training, clear communication and a signed Patient's Informed Consent Form along with the appropriate documentation of any procedure carried out may provide professional safety to practicing obstetricians and gynecologists in case of medical litigation. PMID- 15887610 TI - Asserting the right to life--a challenge to medical guidance on withdrawing treatment. AB - This paper analyses the English High Court decision of R (on the application of Burke) v General Medical Council and considers its implications for the future. Mr. Burke, relying on common law and the European Convention on Human Rights, succeeded in a challenge to the General Medical Council's Guidance on Withholding and Withdrawing Life-prolonging Treatment. He asserted his right to choose to be given treatment so as to live and condemned the Guidance as centred on the right to refuse treatment. PMID- 15887611 TI - Coercion in the treatment of anorexia nervosa: clinical, ethical and legal implications. AB - Because of its high mortality and treatment resistence, clinicians sometimes invoke the law in aid of retaining their most acutely ill-patients in treatment or re-feeding programs. Depending on the jurisdiction, various laws, including mental health and adult guardianship laws, have been invoked to achieve this objective (Carney, Tait, Saunders, Touyz & Beumont, 2003). Until recently, little was known about the therapeutic impact of coercion on patients (Saunders, 2001, Carney & Saunders 2003), or the relative advantages of different avenues of coercion (Carney, Saunders, Tait, Touyz & Ingvarson 2004). Most obscure of all, however, has been our understanding of the factors influencing clinical decisions within specialist anorexia treatment units regarding which in-patients will be selected for coerced treatment. This paper reports legal and ethical implications of findings from analysis of data gathered from a major Australian specialist anorexia treatment facility over nearly 5 years. PMID- 15887612 TI - SUDEP--whether to tell and when? PMID- 15887613 TI - Protecting patients' rights in New Zealand. AB - For 30 years, medical malpractice litigation has effectively been barred in New Zealand by accident compensation laws. Victims of medical error are rehabilitated and compensated through a no-fault, state-funded compensation scheme. Patients' rights (and providers' duties) are set out in a Code of Consumers' Rights, which applies to all providers of health and disability services, ranging from surgeons to traditional Maori healers and unpaid caregivers. Organisational providers such as hospitals are also covered. Complaints alleging a breach of the Code are resolved at the lowest appropriate level by a statutory Ombudsman, the Health and Disability Commissioner. Resolution techniques include advocacy, mediation and investigation, which may result in a finding of breach of the Code. The emphasis is on "resolution, not retribution" and "learning, not lynching". The Commissioner makes remedial recommendations, but does not award damages. Competence concerns are referred to professional registration boards, and serious breaches are referred for disciplinary proceedings. The New Zealand model of legislating patients' rights and enforcing them via an independent Commissioner is an interesting variant of professional self-regulation, and seeks to promote resolution of individual complaints and systemic improvements in health care quality. Investigation reports are widely distributed to promote learning. Despite initial professional resistance, the Office is becoming recognised for its key role in a regulatory system where medical malpractice claims are barred. A case study involving misdiagnosis of a fatal heart attack illustrates the flexibility of the New Zealand response to medical errors. PMID- 15887614 TI - Method of questioning and the accuracy of eyewitness testimony. AB - System variables are integrally part of factors that can be controlled by the legal system to enhance the accuracy of testimony by eyewitnesses. Apart from examining the relationship between questioning as system variable and the accuracy of testimony, the present study furthermore explores the relationship between type of questioning and certain biographical variables (occupation, age, gender and race). To achieve the aim of the study, 412 respondents consisting of 11 to 14-year-olds, university students, the public and Police College students participated and were exposed to open-ended or closed-ended questions. It was found that the participants who responded to the closed-ended questions were significantly more accurate than those who answered the open-ended questions. All the biographical groups, except the public, were more accurate in responding to the closed-ended questions. The scholars obtained the lowest scores (although not always significant) for both the open-ended and closed-ended questions. With respect to age the 18 to 25-year-olds obtained significantly higher scores than the other groups for the closed-ended questions. Whites performed significantly better than blacks in response to the open-ended and closed-ended questions. PMID- 15887615 TI - Psychotherapy ethics with violence victims. AB - There are many special issues that therapists will face while providing psychotherapy services for victims of violence. The primary goal of such intervention must be to reempower the victim so that she perceives herself as the survivor she must become. To do this, she has to deal with the trauma, integrate it into her past, and then, get on with her life. Various problems could occur during custody evaluations, forensic consultations and media exposure. Monitoring confidentiality issues when working with this kind of victims may be crucial to prevent placing them in any further danger. Given the special vulnerability of violence victims, it is essential for the therapist to act in an ethical manner at all times. PMID- 15887616 TI - Involuntary outpatient commitment: a naturalistic study of its use and a consumer survey at one community mental health center in Israel. AB - The following study assessed the efficacy of involuntary outpatient commitment in promoting treatment adherence and preventing hospitalization during and after a period of compulsory treatment in one community mental health center in Israel and evaluated the attitudes of patients and psychiatrists. All patients referred for involuntary outpatient commitment over a two year period (N = 26) were followed up for 3-5 years. After the termination of the commitment, the patient and psychiatrist were interviewed. Although only three patients had been in regular outpatient treatment before the commitment, fourteen remained in regular care during the commitment's duration and ten remained for a further two years. Comparing the follow-up period with the identical duration before the commitment, the number of hospitalizations and number of days hospitalized were significantly reduced. Fifteen were rehospitalized during follow-up. Thirteen patients were interviewed at the termination of the commitment, and nearly all perceived the commitment in negative terms, although eight did not think it had a negative effect on their relationship with the psychiatrist and six thought their opinions were respected. Six of the psychiatrists considered they were not giving optimum treatment. Despite the presence of the commitment, psychiatrists were often sensitive to patients' wishes and the involuntary aspect of the care was often not pursued. While half of the sample were lost to community care, half remained for several years. The study reflects the delicate nature of involuntary care in the community, so that in practice it is often not involuntary, not for outpatients and not a commitment. PMID- 15887617 TI - The forensic nurse practitioner role (emergency nursing)--potential response to changing health needs in New Zealand. AB - There is little existing policy related to Forensic Evidence collection in the Emergency Department environment within New Zealand (NZ). A case study based on the Christchurch Hospital Emergency Department (ChCh ED) environment is presented, which outlines the need to develop a specialised nursing role, that of Forensic Nurse Practitioner (emergency care). The role of nursing input in the field of Forensic Medicine in NZ is essentially limited to the psychiatric focus. While roles such as those of the Forensic Psychiatric Nurse are relatively well established, there is an apparent absence of other 'forensic' functions, in particular those related to victims or perpetrators of crime. It is an accepted feature of emergency care that serious injuries and deaths do occur despite intervention. In addition, there are significant numbers of patients who present as a result of assault or in suspicious circumstances. Awareness of the importance of forensic evidence collection, appropriate storage and disposal of material is growing. Legal implications have significance for nurses, in particular with the movement towards Advanced Nursing Practice with its focus on increasing autonomy, accountability and independent practice. In order to achieve holistic healthcare and to provide appropriate and effective interventions, forensic emergency nursing skills need to be developed. PMID- 15887618 TI - Nursing ethics. PMID- 15887619 TI - Abortion legislation in Eritrea: an overview of law and practice. AB - This article discusses legal issues related to the abortion provisions of the Transitional Penal Code of Eritrea. As is the case in many African countries, the current abortion law of Eritrea mainly was adopted from continental Europe four decades ago, reflecting the reality of the time. Despite the advancement in science and technology, which significantly determines the very definition and concept of abortion and contraception, the abortion law remains the same, save for minor amendments taken place in 1991. Due to the background of the abortion law and the shortcomings occurred during the amendment process, the law manifests legal gaps and limitations resulting in discrepancies between law and practice. The article, therefore, identifies and analyses the gaps of the abortion law in light of principles of criminal law, existing medical technology related to abortion, and experience of other countries. PMID- 15887620 TI - Medically-assisted procreation in Italy. AB - In February 2004, a law was approved in Italy on medically assisted procreation. The authors attempt to discuss the main provisions of the law. In its 18 articles, the law provides access to approved assisted procreation methods for adult couples of different sex, either married or cohabiting, of a potentially fertile age, both living, and establishes that the couple must be offered the chance of reverting to adoption or foster care procedures pursuant to the law in force. The law contains numerous prohibitions and restrictions: heterologous insemination, cloning, the cryo-preservation of embryos and research on human embryos are forbidden; during an assisted reproduction cycle, the methods used must not produce a number of embryos greater than those strictly necessary for a single and simultaneous implant, and in any case never more than three. The law calls for the woman and her husband or partner to be properly informed and for their approval to be given in writing. Fairly severe sanctions have been established for persons who break the law; health workers are however entitled to object on conscientious grounds to performing certain procedures. PMID- 15887621 TI - Organ transplantation in Malaysia: a need for a comprehensive legal regime. AB - Organ transplantation has become increasingly routine as a means of saving and improving the quality of lives of thousands of people each year. However, transplant activity is increasingly constrained by the shortage of organs. The major impediment in procuring organs for transplant in Malaysia is the lack of cadaveric donors. The lack of cadaveric donors has encouraged patients to go to countries like India and China to purchase organs especially kidneys for transplantation. The inadequacies of the existing Malaysian Human Tissues Act 1974 has also contributed to this problem. For instance, the word 'tissue" is not defined under the Act. This raises complex and ethical questions as to the scope of the definition for "tissue". There is also no definition of "the person lawfully in possession of the body." This is significant as he is the person who is empowered by the Act to authorise removal of tissue. Further, there is also no articulation of a hierarchy of relatives who are deemed the next of kin. In a situation involving a large number of relatives, asserting different opinions, this may pose a problem. The articulation of a priority list is particularly difficult in Malaysia as it is a multi-cultural society where the hierarchy of relatives with the right to claim decision-making powers may vary in different cultures. Furthermore, there is also a pressing need for a legislation to ensure that the rights of potential live donors are protected. At the moment, the Human Tissues Act 1974 only relates to cadaveric donors whereas live donors fall within the purview of the common law. The system of "opting out" should be considered in Malaysia whereby every individual is presumed to be a donor unless he or she registers an objection. But this system can only be fair if every person in the community is given notice of the law and understands its implications. For the system to work, there must also exist a simple and effective way of registering objections. There is a need for continuous intensive public education and counselling. A nationally co-ordinated mechanism must be in place to ensure effectiveness of identifying potential donors and recipients. PMID- 15887622 TI - Commercialization of human organs for transplantation: a view from South Africa. AB - The Human Tissue Act 65 of 1983 regulates all aspects regarding organ transplants. This Act was last amended in 1989. Since then medical science has developed tremendously and to such an extent that organ transplants today are almost routine operations in many hospitals. Unfortunately the current methods of procuring human organs are not supplying the demand. A new approach, the commercialization of human organs for transplantation is a possibility with the potential to supply one hundred per cent of the demand for organs. There are however many arguments against the commercialization of human organs, but not one of these arguments is without criticism. Ethical aspects concerning commercialization of human organs also need to be investigated, in order to reach a conclusion that it is not unethical and is worth being investigated. PMID- 15887623 TI - Not just euthanasia: recognizing a legal positive right to palliative care. AB - As jurists, doctors, and ethicists are increasingly engaged in the debate as to whether and how to legalize euthanasia as a matter of public policy, less debate takes place on what it is submitted ought naturally to come before, i.e. the affirmation of a legal right to palliative care. In Israel, the draft law The Terminally Ill Patient (2001) includes a palliative care provision. It is submitted that such provision, laudable as it is, appears within a euthanasia oriented framework and is given too limited a scope. Hence the suggestion that palliative care be given an autonomous role, and be recognized as a positive legal right. PMID- 15887624 TI - Strain variability in Zucker rats affects their response to oral oleoyl-estrone. AB - There is a considerable variability in the responses of Zucker fa/fa rats in metabolic studies, which could not be solely attributed to the leprfa mutation. In order to fathom the extent of this variability, we compared the response to oleoyl-estrone (OE), a powerful lipid-mobilising agent, of two strains of Zucker lean and obese rats: Harlan (H) and Charles River (CR). Rats were given an oral gavage of 10 micromol/day/kg of OE in sunflower oil, and were compared with oil receiving controls. Body composition, energy and water balances, and plasma parameters were studied after 10 days of treatment. H rats showed a higher water turnover than CR rats; OE treatment reduced water intake, partly compensated by metabolic water, and decreased stool water. H rats accrued more cholesterol than CR animals, which showed higher cholesterolaemia. OE facilitated cholesterol disposal in lean (CR and H) and H obese rats. CR rats had higher body and liver lipids than H animals. No differences in energy balance were found. Insulin decrease following OE treatment was greater in lean CR than in H rats, but this trend was reversed in the obese rats, lacking effective responses to leptin. The red cell glucose compartment was smaller in H than in CR rats; the higher insulin levels in H rats may be partly responsible for that difference. Obese H maintained glycaemia (and liver glycogen) with higher insulin levels than CR animals. The extent to which the leprfa mutation affects the responses of Zucker fa/fa rats could not be singled out unless the metabolic environment of the batch used is known. This variability must be taken into account when developing a metabolic or hormonal study in which this model of obesity is used. PMID- 15887625 TI - Microalbuminuria, brain vasomotor reactivity, carotid and kidney arterial flow in Type 2 diabetes mellitus. AB - Diabetic microangiopathy produces widespread small vessel impairment which particularly affects renal glomeruli functions. Microalbuminuria is the earliest marker of microangiopathic kidney disease and has also recently been recognised as a marker of macroangiopathic cardiovascular involvement. To determine correlations between daily microalbuminuria, local microangiopathic kidney damage, systemic macroangiopathic involvement and functional brain microcirculation, 70 Type 2 diabetic subjects who were diagnosed more than 5 years ago underwent carotid (to determine index of macro- and microangiopathy) and interlobar kidney artery color Doppler (to determine microangiopathic involvement), transcranial Doppler (to determine alterations in brain vasomotor reserve), and evaluation of daily albumin excretion rate. All the indices of microcirculatory involvement in the kidney, brain and small vessels downstream from the carotid arteries were closely related (for all p<0.001) but never correlated with the macroangiopathy index. Daily microalbuminuria correlated with all the micro- (p<0.0001) and macroangiopathic (p<0.005) Doppler indices. These findings confirm that microangiopathy is the main cause of the diabetic increase in the albumin excretion rate and support the view that microalbuminuria can be considered a powerful biomarker of widespread macroangiopathy. Our results suggest microalbuminuria may also identify cerebrovascular diabetic involvement, as it predicts both macroangiopathic carotid alteration and microvascular brain impairment. PMID- 15887626 TI - The relationship between levels of alpha1-acid glycoprotein and metabolic parameters of diabetes mellitus. AB - The aim of this study was to investigate the relationship of alpha1-acid glycoprotein (AGP, orosomucoid), which is an acute-phase reactant, in patients with Type 1 (T1DM) and Type 2 diabetes mellitus (T2DM), with the metabolic parameters and complications of diabetes mellitus. A total of 119 diabetic patients (89 T2DM, 30 T1DM) and 33 healthy controls were included in the study. The mean AGP level in the diabetic group was not different from the control group (82.4 +/- 28.9 and 81.6 +/- 16.6 mg/dl, respectively), but it was higher in T2DM than in T1DM subjects (86.6 +/- 28.3 and 69.7 +/- 26.9, respectively). AGP plasma levels showed a significant correlation with age and body mass index (r = +0.348 and r = +0.296, respectively). AGP plasma levels resulted higher in obese diabetic patients (97.7 +/- 28.0) than in non-obese diabetic patients (77.6 +/- 28.7 mg/dl) and controls (81.6 +/- 16.6), and also higher in diabetic subjects with poor glycaemic control (85.1 +/- 33.3 mg/dl) than in diabetic subjects with optimal glycaemic control (79.5 +/- 23.1 mg/dl). A relationship between AGP and macro- and microvascular complications of diabetes mellitus was not found. In conclusion, inflammatory findings were more prominent in obese diabetic patients. PMID- 15887627 TI - Effects of metformin on glucagon-like peptide-1 levels in obese patients with and without Type 2 diabetes. AB - Metformin has been shown to increase glucagon-like peptide-1 (GLP-1) levels after an oral glucose load in obese non-diabetic subjects. In order to verify if this effect of the drug was also present in obese Type 2 diabetic patients who have never been treated with hypoglycemic drugs, 22 Type 2 diabetic and 12 matched non diabetic obese patients were studied. GLP-1 was measured before and after a 100 g glucose load at baseline, after a single oral dose of 850 mg of metformin, and after 4 weeks of treatment with metformin 850 mg three times daily. Post-load GLP 1 levels were significantly lower in diabetic patients. A single dose of metformin did not modify GLP-1 levels. After 4 weeks of treatment, fasting GLP-1 increased in diabetic patients (3.8 vs 4.9 pmol/l; p<0.05), while the incremental area under the curve of GLP-1 significantly increased in both diabetic [93.6 (45.6-163.2) vs 151.2 (36.0-300.5) pmol x min/l; p<0.05] and non-diabetic [187.2 (149.4-571.8) vs 324.0 (238.2-744.0) pmol x min/l; p<0.05] subjects. In conclusion, GLP-1 levels after an oral glucose load in obese type 2 diabetic patients were increased by 4 weeks of metformin treatment in a similar fashion as in obese subjects with normal glucose tolerance. PMID- 15887628 TI - Comparative in vitro uptake of zinc by erythrocytes of normal vs Type 2 diabetic individuals and the associated factors. AB - Diabetes is an oxidative stress-related disorder in which erythrocyte zinc uptake may vary as compared to healthy individuals. Since zinc is one of the important antioxidant trace metals, some functional indices of erythrocyte zinc status, ie in vitro zinc uptake, osmotic fragility and glucose uptake, were compared in Type 2 diabetic subjects (n=43) and healthy controls (n=22). The associations of these indices with plasma levels of antioxidants and micronutrients were examined. The trace metals were analyzed by atomic absorption spectrophotometer. Vitamins were estimated using spectrophotometric and spectroflourometric methods. In vitro zinc uptakes of healthy subjects were 17 to 52% higher (p<0.01) than those for diabetic subjects. The osmotic fragility for diabetic cells was 2.2 to 1.5 times higher than the healthy cells in 0.85-0.5% NaCl solutions (p<0.05). Percent hemolysis at 0.75, 0.65 and 0.55% NaCl had significant negative correlations (p<0.05) with in vitro zinc uptakes and that at 0.50% NaCl had a positive correlation with HbA1c levels (p<0.05). The in vitro zinc uptakes of erythrocytes in healthy subjects showed a strong negative correlation (p<0.01) with percent hemolysis at 0.75, 0.65 and 0.55% NaCl, a positive correlation with plasma zinc (r=0.33, p<0.05) and a strong negative correlation with plasma selenium and iron, hemoglobin and serum ceruloplasmin indicating antagonistic behavior of copper, iron and selenium with zinc uptake (p<0.01). Furthermore, erythrocyte super oxide dismutase (SOD), plasma ascorbic acid and status of riboflavin and thiamine were negatively correlated with in vitro zinc uptakes of erythrocytes in healthy subjects (p<0.01). These associations in the diabetic subjects were weaker than normal. Erythrocyte zinc uptake and osmotic fragility could be biomarkers of long term zinc status and decrease of zinc uptake may be one of the features of diabetic patients. PMID- 15887629 TI - Effects of two different glibenclamide dose-strengths in the fixed combination with metformin in patients with poorly controlled T2DM: a double blind, prospective, randomised, cross-over clinical trial. AB - A double-blind, prospective, randomised, cross-over clinical trial was performed comparing a glibenclamide (G) 5.0 mg/metformin (M) 400 mg combination with a G 2.5 mg/M 400 mg formulation to evaluate whether a higher dose of glibenclamide was able to improve glycaemia in poorly controlled Type 2 diabetic patients. One hundred and ninety-eight patients with poorly controlled Type 2 diabetes mellitus were randomised to receive one of the two trial drugs for a first 3-month period, and were then assigned to the alternative combination for further 3 months. The starting dose (2 tablets/day, 30 min before breakfast and dinner) was to be up titrated to 3 tablets/day when required. A standard dietary regimen was kept constant for the total trial duration. Fasting plasma glucose, HbA1c, C-peptide, insulin and lactate levels, haematology and blood chemistry were measured at the start/end of each cycle. Patients' self-assessment of the glycaemic profile (at fasting and 2 hr after the main meals) was performed weekly. Patients were constantly monitored for adverse events and episodes of hypoglycaemia, and all events were recorded. Decrease of mean fasting glucose levels measured in the first cycle was more pronounced in the group treated with G 5.0 mg/M 400 (p<0.01) compared to baseline, although the difference was not significant--no changes were observed in the second 3-month period. Results of patients' self-assessment of the glycaemic profile in the overall 6-month period show that the two trial drugs produced similar effects on fasting glucose, but the decrease of post prandial glycaemic levels was markedly higher with G 5 mg/M 400 mg than with G 2.5 mg/M 400 mg at both main meals. A similar significant decrease (p<0.01) of HbA1c was observed in both sequence groups at the end of the first 3-month treatment period, and mean levels remained unchanged at 6 months. Drug-related adverse events were observed in 2 patients during treatment with G 2.5 mg/M 400 mg and in 5 with G 5 mg/M 400 mg, while 14 and 22 episodes of hypoglycaemia occurred with the two trial drugs, respectively (p=NS between treatments). Metformin-induced increases of lactate levels were similar in the two sequence groups. No differences between groups were found either in the number of up titrated patients or in all the other laboratory parameters. In conclusion, the new combination containing 5-mg glibenclamide produced a greater improvement in post-prandial glycaemic control compared with the standard fixed doses, and resulted equally safe and well tolerated. PMID- 15887630 TI - Epidemiology of diabetes in pregnancy: a review of Italian data. AB - The prevalence of diabetes and impaired glucose tolerance in women of childbearing age is increasing worldwide, particularly among minority youth. As a consequence, in many parts of the world the number of pregnancies complicated by Type 2 diabetes is actually exceeding those complicated by Type 1 diabetes. Moreover, also the prevalence and incidence of gestational diabetes mellitus have been rising over recent years. Although diabetes complicating pregnancy is a cause of maternal and foetal complications, its exact prevalence in pregnancy is yet not well defined and large population studies are still lacking. This article reviews the existing epidemiologic studies on diabetes in pregnancy performed in Italy. PMID- 15887631 TI - Comparision of interleukin-6 and malondialdehyde levels in diabetic patients in Taiwan. AB - The purpose of this study was to compare the levels of interleukin-6 (IL-6) and malondialdehyde (MDA) in healthy controls and diabetic patients without coronary heart disease (CHD). Fasting serum IL-6 and MDA were determined in 30 healthy controls and 52 (20 Type 1 and 32 Type 2) diabetic patients without clinical evidence of CHD. MDA was calculated as oxidative stress. The IL-6 concentration was used to evaluate the cytokine function. Results showed that the serum IL-6 and MDA concentrations are significantly higher in Type 2 diabetic patients (p<0.05). In conclusion, we demonstrated that Type 2 diabetic patients are more influenced by oxidative stress than Type 1 diabetic patients and healthy controls, because of the action of cytokine. PMID- 15887632 TI - Serum sialic acid and carbohydrate-deficient transferrin concentration in Type 2 diabetes mellitus with and without macrovascular complications. PMID- 15887633 TI - Italian Society of Diabetology Mentor Award. The stages of Type 1A diabetes: retrospective and prospective. AB - In 1986, enough information was available based on studies by investigators from four continents to propose a model of the pathogenesis of Type 1A diabetes. The model divided pathogenesis into a series of stages with the proposal that insulin dependent diabetes was a chronic autoimmune disorder. Long-term studies of non diabetic identical twins of patients with Type 1A diabetes indicated that chronic loss of the ability to secrete insulin preceded the onset of diabetes in the presence of both genetic susceptibility and expression of anti-islet autoantibodies. It is now clear that the great majority of individuals who develop Type 1A diabetes show both immunologic and metabolic abnormalities before the development of overt diabetes. A combination of detection of anti-islet autoantibodies, genetic characteristics and metabolic abnormalities can now be used to predict Type 1A diabetes as well as approximate timing of diabetes onset. This knowledge has led to clinical trials on the prevention of Type 1A diabetes that hope to harness a remarkable increase in basic immunologic knowledge and a new generation of immunomodulatory therapies. Despite this increase in knowledge of the natural history of Type 1A diabetes and pathogenesis in animal models, much remains to be defined to allow efficient and successful disease prevention. PMID- 15887634 TI - Information systems for the materials management department: stand-alone and enterprise resource planning systems. AB - Materials management information systems (MMISs) incorporate information tools that hospitals can use to automate certain business processes, increase staff compliance with these processes, and identify opportunities for cost savings. Recently, there has been a push by hospital administration to purchase enterprise resource planning (ERP) systems, information systems that promise to integrate many more facets of healthcare business. We offer this article to help materials managers, administrators, and others involved with information system selection understand the changes that have taken place in materials management information systems, decide whether they need a new system and, if so, whether a stand-alone MMIS or an ERP system will be the best choice. PMID- 15887635 TI - Hazard report: unreliable alarm notification by StatView paging system could be life threatening. PMID- 15887636 TI - Hazard report: don't get burned: fire risk with liquid ingress into OR tables. PMID- 15887637 TI - Facility design and safety issues for staff and obese patients. PMID- 15887638 TI - [Risks of unsafe (homo)sexuality. Alarming news about a supervirus justify increased preparedness]. PMID- 15887639 TI - [Consequences of violence--a public health issue. Strong connection between violence/threat and illness in both women and men]. AB - Earlier studies have indicated that patients with chronic pain often have experienced physical abuse, but only a small number of population studies have demonstrated an association between abuse and chronic pain. During 1997 a population study was made in northern Sweden. Close to 15,000 people from 18-85 years of age answered a questionnaire about public health issues, including four questions about physical abuse and threats. In the present study the association between answers to these four questions and the other health questions were analyzed for men and women from 18-44 years. Almost 8% of the women and 10% of the men had been subjected to violence or threats during the past year, with higher figures for younger men and women. A strong association between violence/threats and most of the other health issues was found for both men and women. The association remained after controlling for various socioeconomic variables as well as smoking, although with somewhat lower odds ratios. PMID- 15887640 TI - [More violent assaults reported to the police and increasing gang violence. A survey of violence-related personal injuries in Umea]. PMID- 15887641 TI - [Temporal arteritis]. PMID- 15887642 TI - [New investigations in suspected Creutzfeldt-Jakob disease. Analysis of 14-3-3 protein and T-tau in cerebrospinal fluid for safer diagnosis]. AB - Creutzfeldt-Jakob disease (CJD) is a prion disease characterized by rapid neurodegeneration that leads to the death of the patient within months to a few years. Since the disease is transmissible, there is an obligation in Sweden to report possible CJD cases to the Swedish Institute for Infectious Disease Control. To make a diagnosis of CJD is difficult, especially early in the course of the disease when the clinical features may be very vague and heterogeneous. Hence, accurate biological markers both for confirming and excluding CJD would be of great value. The currently recommended investigation of a patient with possible CJD comprises clinical evaluation. electroencephalography, computed tomography or magnetic resonance imaging of the brain and test for 14-3-3 protein in the cerebrospinal fluid (CSF). Recent studies suggest that analysis of total tau (T-tau) and phospho-tau (P-tau) in CSF is a valuable complement to this set of investigations. Here, we review how CSF T-tau and P-tau may aid in the diagnosis of CJD and illustrate this by presenting cases from routine clinical practice. PMID- 15887643 TI - [New European rules for clinical trials competition-promoting for Sweden...can we keep the advantage?]. PMID- 15887644 TI - [National contact tracing of tuberculosis should be introduced]. PMID- 15887645 TI - [Protect the personal integrity!]. PMID- 15887646 TI - [HSAN must have high demands on itself]. PMID- 15887647 TI - [Clinical preceptorship for medical students must be improved]. PMID- 15887648 TI - [Does Jarhult mean that case volume is not significant?]. PMID- 15887649 TI - [Working routines in connection with sick listing must be renewed]. PMID- 15887650 TI - Inhibition of compound action potentials following administration of dental self etching primer/adhesives. AB - Self-etching dentin adhesive systems, a new generation of dental-bonding agents, were investigated for their ability to block nerve conductance in the rat sciatic nerve model. Clearfil SE Bond, Mac Bond and FL Bond were used as test materials. Isolated rat sciatic nerves obtained from female albino rats were placed between two electrodes in a bath containing Tyrode solution. The bonding agents were brought into contact with the nerves and the evoked compound action potentials were recorded versus time before and after contact with the materials. Statistical analysis was performed using the Friedman and Wilcoxon Signed-Ranks tests (P = 0.01) to compare the mean percentage of inhibition between test materials versus time. All test materials were able to block nerve transmission rapidly (3.3 +/- 0.5, 5.1 +/- 2.3 and 8.0 +/- 1.9 min for SE Bond, Mac Bond and FL Bond, respectively). Significant differences were found between FL Bond and SE and Mac Bond (P < 0.01). These results indicated that the bonded systems tested, intended primarily as dentinal tubule occluders, may also exert some direct desensitizing effect on intratubular or pulpal nerve terminals in extremely deep cavities or pulpal microexposures, contributing to decreased postoperative pain. PMID- 15887651 TI - LF-NMR and multivariate data analysis: compression of data to classify hydrogel contact lenses. AB - Hydrogel contact lenses swollen in viscoelastic artificial tears solution have been studied, measuring transversal relaxation times of water molecules using LF NMR techniques. Data were processed by classical multiexponential fitting, by principal component analysis and by SLICING, a multi-way analysis method. The reason for using multivariate data analysis was not to obtain a better fitting, but rather more effective data description. The single-sample relaxation curves were projected in a space spanned by the loading curves, and in this space it was simpler to compare data. In particular, it contributed to the description of the variability of motion characteristics of the water molecule 'families' contained in the studied samples. Applying multivariate techniques, we were able to group lenses with different Equilibrium Water Content (EWC) and with the same water content but different compositions. Accordingly, we were able to point out that, if the lenses are swollen first in physiological solution and then in viscoelastic artificial tears solution, hydration characteristics remained unchanged in all the studied samples, except for 38% EWC lenses. PMID- 15887652 TI - Passive adsorption of human antirrabic immunoglobulin onto a polystyrene surface. AB - The latex agglutination immunoassay technique uses polymer colloids as carriers of adsorbed proteins to enhance the antigen-antibody reaction. The aim of the present work is to study the adsorption of Human Antirrabic Immunoglobulin (HA IgG) on polystyrene latex (PS). The physical adsorption of HA-IgG on PS latex was investigated as a function of pH at 2 mM ionic strength. The amount of HA-IgG adsorbed onto PS latex greatly depends on pH; its value showed a maximum in the neighborhood of the IEP of HA-IgG. The electrophoretic method was applied to characterize latex particles. The influence of the amount of HA-IgG adsorbed (J(ads)) on the electrophoretic mobility and zeta-potential values was also studied. PMID- 15887653 TI - Physical and biological properties of a novel siloxane adhesive for soft tissue applications. AB - The aim of this study was to investigate the adhesive properties of an in-house aminopropyltrimethoxysilane-methylenebisacrylamide (APTMS-MBA) siloxane system and compare them with a commercially available adhesive, n-butyl cyanoacrylate (nBCA). The ability of the material to perform as a soft tissue adhesive was established by measuring the physical (bond strength, curing time) and biological (cytotoxicity) properties of the adhesives on cartilage. Complementary physical techniques, X-ray photoelectron spectroscopy, Raman and infrared imaging, enabled the mode of action of the adhesive to the cartilage surface to be determined. Adhesion strength to cartilage was measured using a simple butt joint test after storage in phosphate-buffered saline solution at 37 degrees C for periods up to 1 month. The adhesives were also characterised using two in vitro biological techniques. A live/dead stain assay enabled a measure of the viability of chondrocytes attached to the two adhesives to be made. A water-soluble tetrazolium assay was carried out using two different cell types, human dermal fibroblasts and ovine meniscal chondrocytes, in order to measure material cytotoxicity as a function of both supernatant concentration and time. IR imaging of the surface of cartilage treated with APTMS-MBA siloxane adhesive indicated that the adhesive penetrated the tissue surface marginally compared to nBCA which showed a greater depth of penetration. The curing time and adhesion strength values for APTMS-MBA siloxane and nBCA adhesives were measured to be 60 s/0.23 MPa and 38 min/0.62 MPa, respectively. These materials were found to be significantly stronger than either commercially available fibrin (0.02 MPa) or gelatin resorcinol formaldehyde (GRF) adhesives (0.1 MPa) (P < 0.01). Cell culture experiments revealed that APTMS-MBA siloxane adhesive induced 2% cell death compared to 95% for the nBCA adhesive, which extended to a depth of approximately 100-150 microm into the cartilage surface. The WST-1 assay demonstrated that APTMS-MBA siloxane was significantly less cytotoxic than nBCA adhesive as an undiluted conditioned supernatant (P < 0.001). These results suggest that the APTMS-MBA siloxane may be a useful adhesive for medical applications. PMID- 15887654 TI - Characterization and degradation of functionalized chitosan with glycidyl methacrylate. AB - The synthesis, characterization and degradation of a hybrid chitosan (CTS)/glycidyl methacrylate (GMA) material are reported. These versatile materials (natural-synthetic materials) are potential candidates for dental restoratives. All materials were characterized by infrared spectroscopy (FT-IR), X-ray diffraction and thermal (DSC) analysis. Particular attention was paid to the thermal stability and chemical resistance of the hybrid CTS materials. From dynamical rheological tests, it was concluded that CTS-GMA solutions behave as physical hydrogels. These pH-sensitive gels are an example of stimuli-responsive polymers, also known as 'smart polymers'. PMID- 15887655 TI - The effect of protein-free versus protein-containing medium on the mechanical properties and uptake of ions of PVA/PVP hydrogels. AB - The effect of two simulated biological environments (protein-free and protein containing) on ion uptake and physical properties of PVA/PVP hydrogels were explored in this work. It was found that over the immersion period in both media, wet mass of the hydrogels decreased and compressive moduli increased, likely due to increased polymer content with water loss as the hydrogels equilibrated with water. These changes were independent of polymer content and immersion medium. However, dry mass of the hydrogels increased dramatically when immersed in protein-free medium, changing only moderately in protein-containing medium. The increase in dry mass was attributed to ion uptake from immersion medium, as confirmed by EDXA. We postulate that differences between ion uptake in protein free versus protein-containing medium is likely the result of serum proteins in the protein-containing medium adsorbing to the surface, inhibiting transport of ions into the hydrogel. PMID- 15887656 TI - Immobilization of glucose oxidase onto gelatin for biosensor construction. AB - The properties of a glucose biosensor made by immobilization of glucose oxidase onto gelatin in a layer of electrochemically deposited polyaniline have been investigated. Glucose oxidase was immobilized within gelatin cross-links with chromium(III) acetate. The glucose oxidase biosensor was developed by forming a polyaniline-deposited electrode surface as support for the immobilized enzyme gel, in order to increase its durability. The polyaniline/gelatin/glucose oxidase biosensor has been characterized using chemical and electrochemical methods. Temperature, pH, cross-linking agent concentration, enzyme concentration, kinetic properties, reusability and the effect of electro-active compounds were among the parameters studied. The response time of the glucose oxidase biosensor is 90 s, the detection limit is below 1 mmol/dm3 and the sensor can be used 20 times within a 2-month period without losing its stability. PMID- 15887657 TI - The influence of cellular seeding density in the microencapsulation of hybridoma cells. AB - The aim of this study was to assess the influence of different seeding densities on the function of hybridoma cells (clone 1B5, IgG 2alpha) producing an anti angiogenic monoclonal antibody (mAb), microencapsulated using a high-voltage electrostatic field. Viable cells were microencapsulated in alginate/poly-L lysine/alginate (APA) capsules and maintained in tissue culture. Cellular growth rates, production and release of mAb from the capsules were assessed. This study shows that hybridoma cells survive, proliferate and remain functionally competent for over one month in vitro after microencapsulation in APA capsules generated in an electrostatic field. However, the cell seeding density had to be at least 10(7) cells/ml for the microencapsulated cells to be viable and to produce and release mAb through the capsule membrane. The maximum monoclonal antibody concentration in this culture was 29.1 microg/ml by day 17, with a tendency to increase, but capsule breakage impeded the follow-up of this determination. PMID- 15887658 TI - Surface properties of PEO-silicone composites: reducing protein adsorption. AB - Silicone-based polymers with reduced protein adsorption were successfully prepared by incorporating mono- or bifunctional poly(ethylene oxide) (PEO) derivatives, respectively, into PDMS during rubber formation using classic room temperature vulcanization chemistry. Characterization of the films by water contact-angle measurements and XPS showed that the PEO was present on the film surface, with greater amounts of PEO at the interface modified with monofunctional PEO. Scanning electron microscopy showed the PEO domains segregated into regular zigzag patterns on the PEO-modified surfaces. Significant reductions in the adsorption of fibrinogen, albumin and lysozyme were observed on both PEO-modified surfaces, although the monofunctional PEO surfaces performed much better in this regard. The reductions in protein adsorption were comparable for all three proteins on both surfaces, suggesting that molecular mass of the protein is not a significant factor in determining the magnitude of protein deposition. Western blot studies showed that the adsorption of proteins from plasma to the monofunctional PEO-modified surfaces was also significantly reduced and surprisingly selective, with very few bands noted relative to the control surfaces and those modified with bifunctional PEO. PMID- 15887659 TI - 2004 innaugural address. PMID- 15887660 TI - ECG of the month. Wide-QRS tachycardia in a 58-year-old man with lung cancer. Right ventricular tachycardia. PMID- 15887661 TI - Radiology case of the month. What is the cause of this patient's knee pain? Dorsal defect of the patella. PMID- 15887662 TI - Young woman with jaundice and itching. Bilary cystadenoma. PMID- 15887663 TI - A middle-aged woman with diarrhea. PMID- 15887664 TI - Stent treatment of spontaneous coronary arterial dissection in patients with known coronary arterial disease. AB - We describe spontaneous coronary arterial dissection (SCAD) in 2 diabetic women, aged 55 and 61 years, with known coronary arterial disease and prior myocardial infarcts. In each, the SCAD caused a fresh myocardial infarct, and balloon angioplasty with stenting successfully restored the true lumen of the dissected artery. Delineating the respective roles of stenting, open operative repair, and conservative management in patients with SCAD is a work in progress. PMID- 15887665 TI - Compliance and outcomes: do African Americans with advanced breast cancer fare worse? AB - Among women with advanced breast cancer, African Americans are thought to have a higher mortality rate than Caucasian. A number of factors, including intrinsic tumor biology, have been postulated for this survival difference. If similar treatment were offered, and actually completed, is there a survival difference? The objectives of this study were to determine 1) if the treatment regimens offered to African Americans and Caucasian were the same, 2) when treatment was offered, was compliance a factor, and 3) if the patients were compliant, was the outcome different? A retrospective review of 72 charts of patients with advanced stage breast cancer from 1988-present at our institution and an affiliated hospital was performed. Data collected include 1) stage of disease at diagnosis, 2) treatment offered, 3) compliance, and 4) outcome (disease-free survival and overall survival). African Americans were defined as those patients who described themselves as African American on their hospital face sheets. The treatment regimens offered were the same for both populations. There was not a significant difference in compliance between the African-American and the Caucasian women. There was no significant difference in disease-free survival or overall survival between the African-American and the Caucasian groups. PMID- 15887666 TI - The epidemiology, syndromic diagnosis, management, and prevention of spider bites in the South. AB - Unlike other arthropods, spiders rarely transmit communicable diseases, and thus play a critical role in the ecosystem by consuming other insects that frequently transmit human diseases, such as mosquitoes, ticks, and fleas. There are more than 30,000 species of spiders worldwide, most of which are venomous, but cannot inflict serious bites due to delicate mouthparts and short fangs. Other insect bites, skin infections, and chemical- or physical-agent exposures are often misdiagnosed as spider bites. Approximately 40 species from 4 major genera of spiders in Louisiana and throughout the South, however, can cause severe human envenomings, with dermonecrosis, systemic toxicity, and, rarely, death. Spider bites can usually be prevented by simple personal and domestic measures. Early species identification and specific management may help prevent serious sequelae of spider bites. PMID- 15887667 TI - Diffuse back pain and urological symptoms: recognizing tethered cord syndrome early. AB - Tethered cord syndrome (TCS) is a rare entity in adults, with only about 100 cases being reported in the literature. It is a syndrome in which patients have neurologic, urologic, and orthopedic signs and symptoms. We present a case of a 40-year-old man with back pain and urinary incontinence who was diagnosed with tethered cord syndrome. After the cord de-tethering in the operating room, his pain improved. TCS is an important condition to consider in the differential diagnosis of an adult who presents with back pain and urological symptoms. Imaging studies should be done, and operative treatment may be required to alleviate the symptoms. TCS should be in the differential diagnosis of any patient who presents with back and lower extremity complaints with superimposed urological problems. If TCS is confirmed radiologically by MRI, surgery can be performed with reasonable results provided that the disease process has not progressed. PMID- 15887668 TI - One-year sequelae in patients with West Nile Virus encephalitis and meningitis in Louisiana. AB - West Nile Virus (WNV) infection can be severe and complicated by neuroinvasive disease, such as meningitis and encephalitis. Not much is known about the one year sequelae following a complicated WNV infection. From July to December 2002, the first large outbreak of WNV in the Southern United States occurred in Louisiana. This epidemic resulted in 329 cases of WNV infection including 125 cases of West Nile fever and 204 cases complicated by meningitis and encephalitis. One year later, during the fall of 2003, a telephone survey was conducted to understand the one-year effects of severe WNV infections. WNV case fatality rate was 19.6%. Death certificates of all the deceased were reviewed to verify the mortality rate from WNV infection. The telephone survey revealed that one-year sequelae from severe WNV infections are common and can affect the body and mind. In addition, it can change a person's perception of the state of their health. PMID- 15887669 TI - A case report associating peripheral arterial disease with highly active antiretroviral therapy. PMID- 15887670 TI - [Your patient wants to go to the mountains. You protect him from mountain sickness and altitude edema]. PMID- 15887671 TI - [Hypothermia and frostbite. First measures for victims of cold]. PMID- 15887672 TI - [With diabetes into the mountains. The metabolism derails faster]. PMID- 15887673 TI - [Hepatopathy, pancreatitis, thrombocytopenia in an alcoholic. Then suddenly fail the kidneys]. PMID- 15887674 TI - [This decisive question requires much intuition. Young, athletic, figure conscious -- or having eating disorder?]. PMID- 15887675 TI - [Lipid therapy in risk patients. Do we concentrate too much on LDL cholesterol?]. PMID- 15887676 TI - [Medicine, how it stands in the textbook. Diagnosis perfect -- patient dead]. PMID- 15887677 TI - [Physicians often treat themselves other than their patients. No trust in their own medicine?]. PMID- 15887678 TI - [Pros and cons of early detection of cancer. Recommend the need for preventive measures, but do not pressure the patient]. PMID- 15887679 TI - [Early detection of colorectal carcinoma. Pros and cons]. AB - Apart from a healthy diet and adequate exercise, early detection measures are applied in the prevention of colorectal. The hemoccult test continues to serve as a screening method. It it proves to be positive, a colonoscopic study should always be performed. Using this approach, 20-30% of polyps in the bowel and early cancer stages can be detected and removed as a prophylactic measure. The hereditary nonpolyposis colorectal cancer syndrome (HNPCC) and familial adenomatous polyposis (FAP) yare the most important risk factors for the development of colorectal carcinoma. However, patients with Crohn's disease or ulcerative colitis also have a greatly increased risk and require more intensive bowel cancer prophylaxis. PMID- 15887680 TI - [Pros and cons of early cancer detection in breast carcinoma]. AB - Mammography is an appropriate method for the detection of early forms of cancer of the breast, and for reducing mortality. Whether this actually succeeds depends upon the quality of the early detection strategies employed. Currently the data on reduction of mortality by mammographic screening remain ambivalent. PMID- 15887681 TI - [Early detection of cancer of the prostate. Pros and cons]. AB - Since the introduction of PSA determination for the detection of cancer of the prostate, there have been discussions of the pros and cons of PSA screening for early detection of this cancer. The PSA test used in combination with digital rectal examination has proven to be a superior diagnostic instrument for the diagnosis of malignant prostate disease at an early stage. The German Guideline Catalog for the early detection of carcinoma of the prostate recommends measurement of the PSA from the age of 45 onwards in patients who have first been comprehensively informed about the further proceedings in the event of a positive result. To date, PSA screening has not been shown to lower the disease-specific mortality rate. Two ongoing randomized, multicentre studies (ERSPC and PLCO) are expected to throw more light on the matter. PMID- 15887682 TI - [The incidence of sudden hearing loss is greater than previously assumed]. AB - The global incidence of sudden hearing loss is quoted to be 5-20 new cases/100,000 inhabitants/year. In the opinion of the present authors, the data on which these figures are based, are too old, methodologically questionable and non-transferable. An analysis of data from the compulsory health insurance carriers in Baden-Wurttemberg and the Nordrhein district suggests that an appreciably higher incidence must be assumed for Germany. Statistical processing of the numerical data of all cost carriers is necessary if we are to obtain an accurate incidence for Germany. PMID- 15887683 TI - [General practitioner covers practices of health insurance. How will the physicians be treated concerning compensations?]. PMID- 15887684 TI - [Cost lump sum in Euro and Cent. What is still not included in the EBM book]. PMID- 15887685 TI - [Calcium gaps in osteoporosis patients close. Blister pack simplifies comedication of calcium and bisphosphonates]. PMID- 15887686 TI - [Excellent adjustment with greater action. To pay attention to improving compliance]. PMID- 15887687 TI - [Diagnosis-quiz. bladder emphysema]. PMID- 15887688 TI - [The 10-minute office visit. Untreated acromegaly shortens life expectancy. You must be able to state the suspected diagnosis]. PMID- 15887689 TI - [Look diagnosis. Overnight to muscleman?]. PMID- 15887690 TI - [So that the patients.would be able to move for a long time after first diagnosis of rheumatism. Approach as an emergency!]. PMID- 15887691 TI - [When to begin rheumatism treatment? Initial combination therapy helps fastest]. PMID- 15887692 TI - [Early therapy -- later retirement]. PMID- 15887693 TI - [Three joints swollen, blood sedimentation increased. Methotrexate to be given immediately?]. PMID- 15887694 TI - [Every tenth person has hay fever complaints. To whom do you advise hyposensitization?]. PMID- 15887695 TI - [Specific brief immunotherapy receives MMW Drug Prize 2004. Four pricks expel pollen allergy]. PMID- 15887696 TI - [Instead of not allowing and "cancer diets" for tumor patients. With baby foods, bitter chocolate and supplement nutrition to prevent cachexia]. PMID- 15887697 TI - [To positively exclude organic disorders. How much diagnosis needs irritable colon?]. PMID- 15887698 TI - [Heart failure--yes or no? The third heart sound and markers point the way]. PMID- 15887699 TI - [Diagnosis and therapy of chronic heart failure]. AB - Incidence and prevalence of chronic heart failure are continuously increasing. Today, heart failure of different etiologies represents the most frequent cause of death in industrialized countries. Improvements in heart failure diagnosis and treatment have decreased lethality. In this context, a valid diagnosis, causative therapy of treatable heart failure etiologies, and efficient pharmacological as well as non-pharmacological management strategies are of equal importance. The principal goals of therapy are prevention, maintenance or even improvement of quality of life and the decrease of heart-failure-related morbidity and mortality. PMID- 15887700 TI - [The value of BNP determination in general practice]. AB - The natriuretic peptides BNP and NT-proBNP are synthesized and released dependent of wall tension in the atria and ventricles. The determination of these peptides in a rapid test makes two markers available that closely correlate with the severity of heart failure. Systolic as well as diastolic left ventricular dysfunction are recorded, but a clear differentiation cannot be made by marker determination. Particularly important is the high negative predictive value and the certainty with which heart failure can be excluded when the plasma level is normal. Besides the diagnostic assistance, natriuretic peptides have a high and above all independent value to assess the prognosis of heart failure, acute coronary syndromes and partially of atrial fibrillation. It is possibile to optimize the therapy of heart failure according to the plasma level. PMID- 15887701 TI - [Disease associated with a pronounced subjective pleasure factor. What fun to behave like a maniac now and then]. PMID- 15887702 TI - [Diabetes mellitus--fitness to drive private and commercial vehicles]. AB - For diabetic drivers of private or commercial vehicles and with impaired driving fitness, it is particularly important to avoid accidents. They must be aware of and know how to avoid potential diabetes-related driving problems. This especially applies to hypoglycemia awareness training, recognition of their hypoglycemic symptoms and intensive self-monitoring of blood glucose levels. There are new EU guidelines for driver's licenses. To determine the suitability of a diabetic person for road traffic, a medical evaluation is necessary. Insulin treated diabetics should receive and observe the "Advice for Motorists under Insulin Treatment." PMID- 15887703 TI - [Fee reduction greater than received practice charge. What to do against double fee loss?]. PMID- 15887704 TI - [Side effect of an old health care reform. Your practice is again worth more]. PMID- 15887705 TI - [Thus the portal vein will be freed from high pressure]. PMID- 15887706 TI - [Look diagnosis. Two slit ears]. PMID- 15887707 TI - Is continuous research necessary in management of post cataract surgery endophthalmitis? PMID- 15887708 TI - Unwrapping the human genome. PMID- 15887709 TI - Carotid artery disease and ocular vascular disorders. AB - The carotid artery supplies blood to the structures of the eye; therefore a variety of ocular vascular disturbances can occur secondary to abnormalities in the parent vessels. The ocular manifestations of chronic carotid artery abnormalities may often be subtle and overlooked or they may be acute, where the contribution of the carotid arteries in the aetiology may not be apparent on routine clinical evaluation. Ophthalmologists and neurologists working independently may not recognise the effects of carotid artery disturbances on the eye and missed diagnoses are not infrequent. In our experience of almost a decade in a tertiarycare referral hospital, we have not recorded any patient referred to us from a primary referring physician (either ophthlamologist or neurologist) as a case of ocular manifestation of a carotid artery disease. This brief review highlights the important clinical features, diagnostic modalities and therapeutic options in this group of disorders. PMID- 15887710 TI - Bacterial adherence to polymethylmethacrylate posterior chamber intra ocular lenses. AB - PURPOSE: Bacterial adherence to intraocular lenses (IOLs) has been incriminated in the pathogenesis of postoperative endophthalmitis. Staphylococcus epidermidis is the most common organism isolated. We studied the in-vitro adhesion of Staphylococcus epidermidis to Polymethylmethacrylate (PMMA) IOLs and the effect of duration of exposure to adherence. METHODS: Two groups of 10 IOLs each were incubated in Staphylococcus epidermidis suspension for 2 minutes and 20 minutes respectively. Adhesion of bacterial cells was determined by counting the number of viable bacteria attached to IOLs. RESULTS: The mean bacterial adherence with 2 minutes incubation was 12,889 +/- 7,150 bacteria/ IOL and with 20 minutes incubation was 84,226 +/- 35,024 bacteria/IOL (P< 0.01). CONCLUSION: Our results show that Staphylococcus epidermidis adheres to PMMA IOLs in vitro and the degree of adherence is less for shorter duration of exposure. We conclude that viable bacteria irreversibly adherent to IOLs may play a role in the pathogenesis of postoperative endophthalmitis. Shorter duration of operative manipulation and exposure to contaminating sources may decrease the chances of postoperative endophthalmitis. PMID- 15887711 TI - Planimetric optic disc parameters in normal eyes: a population-based study in South India. AB - PURPOSE: To study the optic disc parameters of normal eyes in a population-based south Indian study. METHODS: One hundred and fifty three subjects from a population-based sample of 1060 included in the Andhra Pradesh Eye Disease Survey (APEDS) were enrolled in the optic disc study. The male-female ratio, the refractive error and distribution of other ocular parameters in the disc study subjects and the APEDS were not significantly different. Magnification corrected morphometry of optic disc photographs obtained by Zeiss telocentric fundus camera was carried out in one randomly chosen eye of each of these 143 subjects. RESULTS: The mean optic disc parameters with the 95% confidence intervals for the distribution were: disc area 3.37 mm2 (2.04 - 4.7), vertical disc diameter 2.12 mm (1.67 - 2.57), vertical cup to disc ratio 0.37 (0.19 -0.55) and neuroretinal rim area 2.8 mm2 (1.76 - 3.84). The disc area, the vertical cup to disc ratio and the rim area showed a normal distribution. The cup to disc ratio correlated with the vertical disc diameter but the association was not strong. CONCLUSION: The disc area, the vertical cup to disc ratio and the neuroretinal rim area are normally distributed in the South Indian population. The normal optic disc parameters would form a basis for future comparisons in different forms of glaucoma. PMID- 15887712 TI - Profile of glaucoma in a major eye hospital in north India. AB - PURPOSE: To study the clinical profile and distribution of various subtypes of glaucoma in a referral practice in North India. METHOD: A retrospective analysis was done of 2425 patients who attended the glaucoma clinic in a tertiary eye-care centre for five years from January 1995 to December 1999. A detailed history was obtained and a thorough examination was performed, including gonioscopy, disc assessment, applanation tonometry and automated perimetry. Diurnal variation of IOP and provocative tests for glaucoma were done where applicable. RESULT: Primary angle closure glaucoma (PACG) was the most common glaucoma subtype. The primary open angle glaucoma (POAG) to the PACG ratio was 37:63. Chronic angle closure glaucoma (CACG) was the most common PACG subtype. The majority of CACG cases were relatively asymptomatic. Male dominance was seen for POAG, juvenile open angle glaucoma (JOAG), CACG, normal tension glaucoma (NTG) and secondary glaucomas. Female dominance was seen for ocular hypertension (OHT), acute or intermittent ACG and developmental glaucomas. The mean age in years at presentation was POAG:60.54 years (males 61.54 years, females 59.01 years) and PACG: 55.13 years (males 57.25 years, females 53.60). The three common secondary glaucomas were: glaucoma secondary to adherent leucoma, aphakic and pseudophakic glaucomas and traumatic glaucomas. Advanced glaucoma was detected in 42 to 53% of patients and bilateral blindness in 8 to 14% of patients in various subtypes. CONCLUSION: Compared to Caucasians, glaucoma patients in North India seem to present nearly a decade earlier and the disease is more advanced at presentation. While PACG is the most commonly encountered glaucoma, NTG and exfoliative glaucoma are relatively rare. PMID- 15887713 TI - Postoperative astigmatism after intracapsular cataract surgery: results of a randomised controlled trial in Nepal. AB - PURPOSE: Postoperative astigmatism following intracapsular cataract extraction with or without anterior chamber intraocular lens implantation is reported as an outcome from a randomised controlled trial. METHODS: Five hundred and two of 1002 eyes randomised to intracapsular cataract extraction with anterior chamber intraocular lens (ICCE/AC IOL) and 417 of 998 eyes to intracapsular cataract extraction with aphakic spectacles (ICCE/AS) were seen for objective refraction one year after surgery. The prevalence and axis of astigmatism were evaluated using univariate analysis. Logistic regression was used to compare the postoperative astigmatism between the groups. RESULTS: Acceptable astigmatism ( 0.5 to 0.0 DCyl) in the AC IOL group was found in 60 (12.0%) patients (95% CI 9.1%-14.9%) and in the aphakic spectacles group (AS) in 69 (16.5%) patients (95% CI 12.9%-20.1%), moderate astigmatism (-1.0 to-1.5 DCyl) was found in 153 (30.4%) patients (95% CI 26.4-34.6%) in ACIOL group and in 288 (69.1%) patients (95% CI 64.6%-73.6%) in AS group; and large astigmatism (-2.0 to - 8.0 D Cyl) was found in 289 (57.6%) patients (95% CI 53.1%-61.6%) in ACIOL group and in 60 (14.4%) patients (95% CI 11.0% 17.8%) in AS group. Large astigmatism was approximately four times more common in the ICCE/AC IOL group compared to ICCE/AS group. In both groups, most patients had "against-the-rule" astigmatism, 446 (88.8%) (95%CI 86.0%-91.6%) in AC IOL group and 348 (83.5%) (95%CI 79.9%-87.1%) in AS group. CONCLUSION: Astigmatism is common after intracapsular cataract extraction. Insertion of an anterior chamber IOL increases the risk of astigmatism. PMID- 15887714 TI - Genetic profile of 81 retinoblastoma patients from a referral hospital in southern India. AB - PURPOSE: To determine chromosomal abnormalities and inheritance pattern in patients with retinoblastoma from a referral hospital in southern India. MATERIALS AND METHODS: Eighty-one retinoblastoma patients from 78 families were included in this study. Peripheral venous blood was taken for chromosomal analysis and pedigree was ascertained for segregation analysis. RESULTS: Male to female ratio was 1.7:1, 55.56% were bilateral retinoblastoma, the mean age of onset was 12.37 months in bilateral and 33.07 months in unilateral cases (p=0.048). Majority (90.12%) had sporadic inheritance and 6.17% had autosomal dominant inheritance. In chromosomal abnormalities, 8.33% had 13q14 deletion, three cases had de novo balanced translocations. CONCLUSION: The age of onset of the disease was much earlier in the bilateral cases compared to unilateral cases. Sporadic inheritance was predominant while only a small percentage of patients had autosomal dominant inheritance. The percentage of patients with 13q14 deletion was higher than reported in the literature and three novel chromosomal translocations were observed. This is one of the largest series of cases reported from India. PMID- 15887715 TI - Occupational open globe injuries. AB - PURPOSE: Occupational ocular trauma is an important cause of acquired monocular blindness in a rapidly industrialising country like India. Knowledge of the epidemiology of occupational eye injuries is essential to formulate viable industrial safety measures. METHOD: We retrospectively reviewed all patients with occupational open globe injuries between 1994 and 1998. We documented the circumstances of the injuries, their clinical findings and the use of appropriate protective eyewear at the time of the injury. The visual acuity 6 months after the injury was the final outcome measure. RESULTS: In this study period we examined 43 patients with open globe injuries sustained at the work place. Thirty four (79.1%) patients were young males. The iron and steel industry accounted for 19 (44.2%) cases while 8 (18.6%) patients each were from the agricultural, mining and other small scale industrial sectors. At the time of the injury, 33 (76.7%) were not wearing the recommended protective eyewear and 6 (13.9%) were under the influence of alcohol. The injuries were mild in 6 (13.9%), moderate in 18 (41.9%) and severe in 19 (44.2%) patients. At the end of 6 months, 2 (4.7%) patients had a visual acuity of 6/12 or better, 4 (9.3%) had a visual acuity of 6/18 to 6/60 and 29 (67.4.%) had a vision of <6/60. Eight (18.6%) patients were not available for follow up. CONCLUSIONS: Occupational open globe injuries are usually severe and are associated with a poor visual outcome. Mandatory use of protective eyewear and alcohol-free environment at the work place is likely to reduce the incidence of severe occupational open globe injuries. PMID- 15887716 TI - Panophthalmitis due to rhizopus in an AIDS patient: a clinicopathological study. AB - Various opportunistic infections in the eye have been reported earlier in AIDS. We report a case of panophthalmitis in an AIDS patient where the eviscerated tissue on histopathologic and microbiologic examination showed the fungus Rhizopus. PMID- 15887717 TI - Malignant melanoma of the choroid with vitiligo. AB - An unusual case of an Asian patient with malignant melanoma of the choroid with vitiligo is reported. Other uncommon features of this case are metastasis to an extrahepatic site without liver metastasis and infiltration of the optic nerve. PMID- 15887718 TI - Vogt-Koyanagi-Harada syndrome following injury-induced progressive vitiligo. AB - We report a case of Vogt-Koyanagi-Harada (VKH), Syndrome wherein the patient developed ocular symptoms following injury-induced progressive vitiligo with immunologic evidence from the skin biopsy specimen of the vitiligo. This case supports the hypothesis that the cell-mediated immune process against a common melanocyte antigen plays an important role in the development and progression of the disease. PMID- 15887719 TI - Frosted retinal branch angiitis in an immunocompetent adult due to herpes simplex virus. AB - A case of acute severe loss of vision due to perivasculitis typical of frosted retinal branch angiitis due to Herpes simplex virus in an otherwise healthy young Omani lady is presented. PMID- 15887720 TI - Prevention of postoperative infections in ophthalmic surgery. AB - Postoperative endophthalmitis is a serious, vision-threatening complication of intraocular surgery. Better instrumentation, surgical techniques, prophylactic antibiotics and better understanding of asepsis have significantly reduced the incidence of this complication. Postoperative endophthalmitis may occur as an isolated event or as a cluster infection. Topical antibiotics, preoperative periocular preparation with povidone-iodine combined with a sterile operating room protocol significantly reduce the incidence of isolated postoperative endophthalmitis. The role of antibiotics in the irrigating fluid and subconjunctival antibiotics remains controversial. Cluster infections on the other hand are more likely to occur due to the use of contaminated fluids/viscoelastics or a breach in operating room asepsis. Prevention of postoperative endophthalmitis requires strict adherence to operating room norms, with all involved personnel discharging their assigned roles faithfully. PMID- 15887721 TI - Locating the retinal break(s) in rhegmatogenous retinal detachment: the first step in successful management. PMID- 15887722 TI - Ocular surface failure is treatable. PMID- 15887723 TI - New perspectives in ocular surface disorders. An integrated approach for diagnosis and management. AB - The cornea, conjuctiva and the limbus comprise the tissues at the ocular surface. All of them are covered by stratified, squamous, non-keratinizing epithelium and a stable tear film. The ocular surface health is ensured by intimate relationship between ocular surface epithelia and the preocular team film. There are two types of ocular surface failure. The first one is characterized by squamous metaplasia and loss of goblet cells and mucin expression. This is consistent with unstable tear film which is the hallmark of various dry-eye disorders. The second type of ocular surface failure is characterized by the replacement of the normal corneal epithelium in a process called limbal stem cell deficiency. It is essential to establish accurate diagnosis for appropriate management of complex ocular surface disorders. There has been considerable advancement in the understanding of the pathophysiology of ocular surface disease. Management has improved with introduction of the limbal stem cell concept and use of amniotic membrane transplantation. PMID- 15887724 TI - Comparison of external dacryocystorhinostomy and 5-fluorouracil augmented endonasal laser dacryocystorhinostomy. A retrospective review. AB - PURPOSE: To compare the success rates of external dacryocystorhinostomy (EXT-DCR) with 5-fluorouracil (5-FU) augmented endonasal laser dacryocystorhinostomy (ENL DCR) and to record the complications associated with 5-FU augmented ENL-DCR MATERIALS AND METHODS: This was a retrospective non-randomised study. Forty-one patients with primary acquired nasolacrimal duct obstruction underwent an EXT-DCR (19 patients) or an ENL-DCR (22 patients) over a 3-year period. A Holmium YAG laser (Ho:YAG) was used in the latter group of patients. Silicone tubes intubated in all patients were removed at three months. 5-FU was applied intraoperatively at the site of the ostium in the ENL-DCR patients. The median follow-up was 12 months (range 3-24 months) for the ENL-DCR group and 22 months (range 6-28 months) for the EXT-DCR group. The patency of the lacrimal system and the severity of epiphora were assessed at a final-review. RESULTS: The median age of the EXT-DCR group was 77 years (range 53-87) and that of the ENL-DCR group was 71 years (range 23 to 84). There were 12 female patients in the former group and 19 in the latter. The percentage of success in the EXT-DCR group was 94.7% (95% confidence interval (CI) = 75.4-99.1) = ), and 63.6% in the ENL-DCR group (95% CI= 43.0-80.3). The confidence interval for the difference of 31.1% was 5.6-52.2. There was a statistically significant difference between the two groups, p=0.024 (Fisher exact test). CONCLUSIONS: These data suggest that EXT-DCR provides better results than 5-FU augmented ENL-DCR. However, ENL-DCR is the procedure of choice in certain circumstances such as in elderly, frail or medically unfit patients. Our results of 5-FU augmented ENL-DCR compare favourably with other published series. PMID- 15887725 TI - Efficacy of topical and systemic itraconazole as a broad-spectrum antifungal agent in mycotic corneal ulcer. A preliminary study. AB - PURPOSE: To evaluate the efficacy of topical (1%) and systemic itraconazole against common fungi such as Aspergillus and other filamentous fungi that cause mycotic corneal ulcer. METHODS: A prospective randomised, controlled study was done in 54 clinically suspected cases of fungal keratitis of which 44 were culture proven. Half the cases (n=27) with superficial involvement were treated with only topical itraconazole (1%) and the other half were treated with both topical and systemic itraconazole. RESULTS: Aspergillus, Penicillium and Fusarium were the most common fungi isolated. The ulcer resolved in 42 eyes (77%) and 12 eyes (23%) did not respond well to treatment. Four of 12 non-responding eyes were caused by Fusarium species. CONCLUSION: Itraconazole, given either topically or systemically, is effective in treating mycotic corneal ulcers. PMID- 15887726 TI - Anterior capsular staining with trypan blue for capsulorhexis in mature and hypermature cataracts. A preliminary study. AB - PURPOSE: To study the efficacy and safety of 0.1% Trypan Blue dye to stain the anterior capsule for capsulorhexis in mature and hypermature cataracts. METHODS: This preliminary study included 25 eyes of 25 patients with a unilateral mature or hypermature cataract, including one case of traumatic mature cataract. In all these cases 0.2 ml of 0.1% trypan blue dye was used to stain the anterior capsule. The efficacy and safety of the dye was evaluated on the basis of intraoperative and postoperative observations. RESULTS: In all 25 eyes the capsulorhexis was completed. There was peripheral extension of the capsulorhexis in the eye with traumatic cataract and the stained edge of the anterior capsule helped identification and redirection of the capsulorhexis. Successful phacoemulsification with intraocular lens implantation was performed in all eyes. Adverse reactions related to the dye such as raised intraocular pressure, anterior chamber inflammation and endothelial damage were not observed in the immediate postoperative period or at the end of mean follow-up of 3 months. CONCLUSION: Trypan blue dye staining of the anterior capsule appears to be a very useful and safe technique that simplifies capsulorhexis in mature and hypermature cataracts. PMID- 15887728 TI - Magnitude of the problem of retinopathy of prematurity. experience in a large maternity unit with a medium size level-3 nursery. AB - This report describes the extent and severity of retinopathy of prematurity (ROP) in a large maternity unit. The screening of 79 preterm babies showed that ophthalmic examinations should become an important part of neonatal care. PMID- 15887727 TI - Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis. AB - PURPOSE: We describe a simple procedure of Immunoperoxidase (IP) technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. METHODS: A high titre polyclonal antibody against a local clinical isolate (axenic) of Acanthamoeba species (trophozoite lysate antigen) was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1) and patients showing no cysts in similar scrapings (pool-2), were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison) with calcofluor white staining and culture. RESULTS: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%). CONCLUSION: The immunoperoxidase technique is a simple and useful test in the diagnosis of Acanthamoeba keratitis. This can supplement the culture results. PMID- 15887729 TI - Ocular fireworks injuries. Clinical features and visual outcome. AB - In a prospective analysis of ocular fireworks injuries over three consecutive years (1997 to 1999) during the Indian festival of Diwali, 42 patients presented with various types of fireworks-related ocular injuries. Twenty two (52%) patients had to be admitted for the management of their ocular injuries. PMID- 15887730 TI - Suprachoroidal haemorrhage. Secondary management. AB - We report a case of successful management of expulsive suprachoroidal haemorrhage following cataract surgery. This case study highlights the strategy of adequate management. PMID- 15887731 TI - Retinitis pigmentosa patients with sickle cell disease and dextrocardia and situs inversus syndrome. AB - Two cases of retinitis pigmentosa (RP) with associated sickle cell disease in one patient, and situs inversus totalis in the other are reported. To our best knowledge, these associations have never been reported in RP. PMID- 15887732 TI - Subconjunctival adult bancroftian filarial worm. AB - We report a case of live adult Bancroftian Filarial nematode removed from the sub conjunctival tissue of a 58-year-old woman from the temporal limbus of her left eye. PMID- 15887733 TI - An unusual case of a retained metallic arrowhead in the orbit and sphenoidal sinus. AB - A case of retained metallic arrowhead in orbit and sphenoidal sinus through an unusual route is reported. The eyeball was removed because of a possible risk of sympathetic ophthalmia. PMID- 15887734 TI - Finding the retinal break in rhegmatogenous retinal detachment. AB - The development of subretinal fluid is governed by a limited number of anatomical factors and gravity. As a result, rhegmatogenous retinal detachments form in a predictable manner around the retinal break of their origin. The shape of the detachments points to the position of the break. The purpose of this review is to describe the characterstic contours of subretinal fluid in rhegmatogenous retinal detachments, and to highlight some rules and methodology which can help in the detection of the retinal break in phakic, psuedophakic and recurrent retinal detachments. PMID- 15887735 TI - Postoperative astigmatism after intracapsular cataract surgery: results of a randomised controlled trial in Nepal. PMID- 15887736 TI - Postoperative astigmatism after intracapsular cataract surgery: results of a randomised controlled trial in Nepal. PMID- 15887737 TI - A prospective study of 413 cases of lens-induced glaucoma in Nepal. PMID- 15887738 TI - ["I accuse"]. PMID- 15887739 TI - [Debate on clinical research without distortion of proportions]. PMID- 15887740 TI - [Independent clinical trials are imperative]. PMID- 15887741 TI - [Multiple sclerosis and physical training]. PMID- 15887742 TI - [Multiple sclerosis: its change in prevalence in Denmark, 1950-1996, and its total prevalence in Danish counties in 1996]. PMID- 15887743 TI - [Excimer laser treatment of recurrent erosion of the cornea]. PMID- 15887744 TI - [New treatment of cystic hygromas/lymphangiomas]. PMID- 15887745 TI - [Documentation of informed consent in medical records--in hospitals and general practice]. PMID- 15887746 TI - [A dose-response study of tranexamic acid. The effect on nausea and blood loss after total knee replacement]. PMID- 15887747 TI - [Self-evaluated health status after cervical cancer surgery]. PMID- 15887748 TI - [Better reporting of harms in randomized trials: an extension of the CONSORT statement]. PMID- 15887749 TI - [End-of-life practices in European intensive care units]. PMID- 15887750 TI - [Klatskin's tumors]. PMID- 15887751 TI - [Schwannoma of the facial nerve--a rare disease, the choice of diagnostic imaging is very important]. PMID- 15887752 TI - [Malingering. The doctor's dilemma]. PMID- 15887753 TI - [Traumatic dissection of the internal carotid artery]. PMID- 15887754 TI - [2004 myoses]. PMID- 15887755 TI - [Photodynamic therapy of senile macular degeneration]. PMID- 15887756 TI - [Mannan binding lectin and spontaneous abortion]. PMID- 15887757 TI - [HbA1 and type 2 diabetes--what is optimal?]. PMID- 15887758 TI - [Guidelines or health]. PMID- 15887759 TI - [The electronic medical record can promote health sector ethics]. PMID- 15887760 TI - [Danish pharmacies have a safe and quick electronic service]. PMID- 15887762 TI - Rural health research: building capacity and influencing policy in the United States and Canada. PMID- 15887763 TI - Registered nurses working alone in rural and remote Canada. AB - This paper describes the demographics of Registered Nurses (RNs) who work alone in rural and remote Canada, their workplaces, and the benefits and challenges of this unique nursing employment situation. Data presented are from a national survey, one of 4 principal approaches used in conducting the project The Nature of Nursing Practice in Rural and Remote Canada. Of the total survey sample, 412 nurses (11.5%) were employed as the only RN in their work setting. Variables of interest included level of education, employment setting, and regional distribution of workplaces. An exploration of predictors of work satisfaction confirmed previous research findings with respect to the importance of continuing education and face-to-face contact with colleagues. Findings from this analysis may inform policy decisions regarding the employment of RNs in rural and remote Canada. PMID- 15887764 TI - Rural diabetes education: does it make a difference? AB - This cross-sectional study using a mailed survey evaluated outcomes of diabetes education, care, and support provided at 2 clinics in rural Manitoba, Canada. Seventy-eight respondents with type 2 diabetes, including 51 rural clinic attenders and 27 non-attenders (mean age = 68.5), were compared regarding demographic characteristics; diabetes education and knowledge; diabetes self management efficacy, attitudes, and behaviours; quality of life; satisfaction with care; and health-care utilization. Attenders had more diabetes education (P < .001), higher self-management efficacy scores (P < .051), and greater satisfaction with diabetes care (P < .001) than non attenders, but more education did not translate into greater diabetes-related knowledge, attitudes, or behaviours. While clinic attendance and supportive interaction with health professionals and peers may help equip people with type 2 diabetes with the confidence and alternative strategies to handle diabetes-related health problems, a more standard approach to diabetes education and care could improve the achievement of desired outcomes. PMID- 15887765 TI - Lives of rural women after myocardial infarction. AB - This study examines the influences of rurality on the lives of women post myocardial infarction (MI). Using a critical ethnographic approach, the researchers analyze in-depth interviews with 12 women from southwestern Ontario, Canada, for the ways in which their experiences were related to social, political, and cultural forces associated with rurality. Data analysis revealed 4 themes: reticence, characterized by a tendency to minimize worry and accept one's life post-MI; referral games, or the challenges associated with accessing tertiary care; resourcefulness in managing one's recovery; and relationships, with rural health professionals and institutions being highly valued. The findings have relevance for nurses in both rural and urban settings who care for women post-MI and form a basis for supporting and building culturally specific post-MI care. PMID- 15887766 TI - Rethinking emancipation and empowerment in action research: lessons from small rural hospitals. AB - A primary goal of action research is social change that is driven largely by the research participants. A major assumption is that through the research process, participants are enabled to take knowledgeable action in their personal, work, or community environments, and that through this action they experience empowerment. Another is that action becomes possible as a result of enlightenment and emancipation through participation in the research. These assumptions were called into question during the course of an interpretive action research study conducted with nurses employed in 3 small rural hospitals in northern British Columbia, Canada. Examination of the issues that emerged during the study illuminates the ways in which nurses' professional and community lives are intertwined. This interconnection provoked a re-examination of how empowerment and emancipation can be realized by nurses in small rural hospitals. PMID- 15887767 TI - The effects of nursing turnover on continuity of care in isolated First Nation communities. AB - Many of Canada's northern First Nation communities experience difficulty recruiting and retaining appropriate nursing staff and must rely on relief nurses for short-term coverage. The latter often are not adequately prepared for the demanding nature of the practice. This study examined the consequences of nursing turnover on the continuity of care provided to residents of three Ojibway communities in northern Ontario. The findings are based on a review of 135 charts of oncology, diabetes, and mental health clients, and on interviews with 30 professional and paraprofessional health-care providers who served the communities. Nursing turnover is shown to detrimentally affect communications, medications management, and the range of services offered; it also results in compromised follow-up, client disengagement, illness exacerbation, and an added burden of care for family and community members. PMID- 15887768 TI - For work and for school: internal migration of Canada's rural nurses. AB - Using data from the Registered Nurses Database and a recently conducted national survey, this study examined the internal migration patterns of Canadian-educated rural RNs. Inter-provincial migration rates, ranging from 11% to 27% depending on the database used, mask much wider variations in sub-provincial movement rates, which are particularly relevant when considering the provision of nursing services in rural and remote communities. Rural RNs are more likely to migrate if they are female, older, working in nursing stations, and living in remote communities. A majority of RNs whose migration is associated with going to school after their initial nursing education do not return to the jurisdiction where they were first registered. Targeted migration studies are needed to fully understand both the detailed patterns and the predictors of such movements in order to better assess recruitment and retention policies and to enhance our overall health human resources planning models. PMID- 15887769 TI - A profile of registered nurses in rural and remote Canada. AB - Research on nursing practice issues in rural and remote areas of Canada is very limited. This report describes the method and initial results of a comprehensive survey of registered nurses (RNs) practising outside the commuting zones of large urban centres, designed to determine: who practises nursing in rural and remote Canada; the nature and scope of their nursing practice; and their satisfaction with their work, community, and practice supports. Using a mailed questionnaire with persistent follow-up, the data-collection frame included a stratified random sample of rural RNs and the full population of RNs who worked in the northern territories and outpost ("remote") settings. The analyses focus on regional comparisons of demographics and primary work settings and on provincial comparisons of satisfaction levels related to work and community. The survey is part of a larger multi-method project intended to inform policy on rural nursing practice in Canada. PMID- 15887770 TI - Innovations and issues in the delivery of continuing education to nurse practitioners in rural and northern communities. AB - This paper addresses the need to provide rural nurse practitioners (NPs) with the distance education that is considered vital to the upgrading of their professional skills. The method of delivering the courses is a critical aspect of their success. The authors trace and describe the innovative delivery of the Rural Ontario Nurse Practitioner Continuing Education Initiative, from the initial needs assessment study through to the implementation and evaluation study. In each study, a multi-method action research model was used. The respondents showed a preference for face-to-face modalities that were perceived to be constrained by barriers. These barriers were subsequently addressed by the pilot project. Those living in rural areas recognized the benefits of information technologies. Implementation was effectively weighted on multiple modes of online course delivery and the use of constructivist pedagogy. The findings suggest that the delivery of continuing education to rural and remote NPs is still wrought with challenges. PMID- 15887771 TI - So, what do we mean by "rural," "remote" and "northern"? PMID- 15887772 TI - Community partnerships: translating research for community development. PMID- 15887773 TI - Dementia care in rural and remote areas: the first year of a CIHR new emerging team. PMID- 15887774 TI - Medicare Advantage gaining traction for 2005 and beyond. PMID- 15887775 TI - Researchers cite five major challenges facing Medicare Advantage plans. PMID- 15887776 TI - Report illustrates detailed analysis used to calculate TennCare per capita costs. PMID- 15887777 TI - Care, disease management will drive HMO profitability, especially for seniors. PMID- 15887778 TI - [Reliability of % vol. declarations on labels of wine bottles]. AB - The Council Regulation (EC) no. 1493/1999 of 17 May 1999 on the common organisation of the market in wine (Abl. L 179 dated 14/7/1999) and the GMO Wine 2000 (Annex VII A) stipulates that the labels of wine bottles have to indicate, among others, information on the sales designation of the product, the nominal volume and the alcoholic strength. The latter must not differ by more than 0.5% vol. from the alcoholic strength as established by analysis. Only when quality wines are stored in bottles for more than three years, the accepted tolerance limits are +/- 0.8% vol. The presented investigation results show that deviations have to be taken into account which may be highly relevant for forensic practice. PMID- 15887779 TI - [Differential diagnosis of traumatic skull findings in infants: residues after vacuum extraction]. AB - If autopsy findings in an infant show traumatic changes on the skullcap, these are not always the result of a postnatal trauma due to child abuse, a fall or another accidental event. With regard to differential diagnosis a birth trauma should also be kept in mind, so that the history of the delivery is important. The spectrum of possible residues after vacuum extraction (circular fracture and/or elevation of the outer table of skull bones, subperiostal and intraossal haematoma, extradural and subdural haemorrhage) is demonstrated by means of three examples from the forensic autopsy material. PMID- 15887780 TI - [Measuring the distance of the wisdom teeth from the occlusal plane as forensic odontological method for chronological age determination]. AB - As part of an epidemiological study on tooth eruption it was attempted to define easily accessible criteria for age estimation in living human beings based on the distance of the wisdom teeth from the occlusal plane. 253 cast models of 149 female and 104 male patients were used for evaluation. Due to the great variability observed in the eruption of the wisdom teeth, only a small fraction of the total number of 10,924 cast models was suitable for investigation. Only teeth with the axis being perpendicular to the plane of occlusion during eruption according to radiological and clinical findings were included. After defining these inclusion criteria, the distance of the wisdom teeth from the plane of occlusion was determined and correlated to the age of the subjects. The results confirmed the known large variability concerning wisdom tooth eruption and did not show any reliable relation between age and distance values. Nevertheless, a remarkable finding that may be relevant for forensic odontostomatology was seen in one case where the lower wisdom teeth were taller than the occlusal plane, although the female patient was younger than 14 years. The upper molars were retained and root development was incomplete. PMID- 15887781 TI - [Homicide by stab to the neck with subsequent attempted suicide by cut to the neck]. AB - A 37-year-old Turkish woman was fatally injured by her 65-year-old Turkish husband with a stab to the cervical region. After her death he inflicted a deep cut to his own neck, which he survived thanks to immediate medical assistance. To the authors' knowledge this combination of a homicidal stab to the neck and a suicidal cut to the neck has not been described before. Kosher butchering, which is common in the couple's traditional cultural environment, is discussed as possible reason for the choice of the neck as site for applying sharp violence. It may have induced the perpetrator to choose this anatomical region. PMID- 15887782 TI - [Complex suicide by two gunshots to the chest and consecutive hanging: circumstances and aspects of reconstruction]. AB - In cases of suicidal gunshots, the capacity to act may be preserved for a certain period of time, so that the suicide may be completed with another method. In the presented case a 74-year-old man was found hanged on the first floor of his house with two gunshot wounds in the chest. Both on the ground and the upper floor traces of suicidal acts were detected. As shots to the precordial region suggest rapid incapacitation, a suicide extending over two floors seemed almost impossible without knowledge of the cause of death and evaluation of the ability to act. The autopsy findings confirmed vital signs of hanging. Moreover, a through-and-through gunshot wound of the chest and abdomen affecting the spleen and a shot lodged in the body without injuring the lung were found. The injury findings thus sufficiently explained the preserved ability to act. The presented case shows characteristics of a complex suicide not yet described so far, but could be clearly classified as suicide in congruity with the pertinent literature. PMID- 15887783 TI - [Determination of butane in respiratory gases by means of GC/MS and GC/MS-MS]. AB - Butane is inhaled in order to achieve a pleasurable state of intoxication. An overdose can lead to death. In two deaths from our own investigation material the circumstances were suspicious for the inhalation of liquid gas, and the presence of butane should be demonstrated in the respiratory gases. For detection, a method of ion trap gas chromatography/mass spectrometry (GC/MS) and tandem-mass spectrometry (GC/MS-MS) was developed, whereby the gas samples from the lung tissue were directly injected into the GC. The GC/MS tests revealed the presence of butane. Moreover, it was found that during the MS-MS tests reaction products appeared which had formed in the ion trap. Systematic investigations of these reaction products showed that these appeared regularly and could be used as additional backup for the proof of butane. Thus phenomena in the ion trap were used which would not have been expected to occur in normal mass spectrometry or tandem-MS and had not been described in the forensic literature so far. The detected amount of butane could be quantified by means of serial dilutions with nitrogen and room air. The described method shows that small molecules or gases can be demonstrated with the ion trap mass spectrometer. PMID- 15887784 TI - Dissatisfaction: how it has grown. PMID- 15887785 TI - Patients in pain: who should be responsible? PMID- 15887786 TI - Pain management. PMID- 15887787 TI - Snowmobile trauma, alcohol and the law: a reply. PMID- 15887788 TI - Primary total knee arthroplasty in patients receiving workers' compensation benefits. AB - OBJECTIVE: To determine the influence of Ontario Workplace Safety and Insurance Board (WSIB) benefits on short-term clinical outcomes of primary unilateral total knee arthroplasty (TKA). METHODS: In a retrospective matched-cohort study at a single tertiary-care arthroplasty centre in Ontario, we compared a study cohort of 38 successive primary TKA patients receiving WSIB benefits from 1998 to 2002 to 38 controls, a matched cohort of non-WSIB patients, comparing Oxford Knee Score and Knee Society Score (both clinical and functional components) as well as flexion and pain variables, preoperatively and at postoperative intervals of 6 weeks, 6 months and 1 year. At least 1 year after their surgery, all patients were asked to complete a non-validated patient satisfaction survey. The number of clinic visits related to the operation was also compared, by means of Ontario Hospital Insurance Plan billing codes for each individual. RESULTS: Preoperative measurements showed the 2 groups to be similar. At follow-up, WSIB patients had significantly higher pain scores, poorer self-perceived functional outcomes and a lower range of knee flexion than the control group. WSIB patients also required more postoperative clinic visits and were more reluctant to answer questions about functional outcome. CONCLUSIONS: Short-term outcomes of primary TKA in patients receiving WSIB benefits are inferior to those of non-WSIB patients. WSIB patients are seen more frequently for postoperative follow-up, which we would attribute to the persistence of subjective complaints after TKA. PMID- 15887789 TI - Multiple epiphyseal dysplasia in children: beware of overtreatment! AB - PURPOSE: To determine the various musculoskeletal manifestations of multiple epiphyseal dysplasia in children, and the course of this disease in childhood. METHODS: Ten children were diagnosed and treated at the Children's Hospital of Eastern Ontario (CHEO) for multiple epiphyseal dysplasia, 1976-2001. We reviewed the clinical, pathological and radiographic records for these cases to determine cause of presentation and progress of the musculoskeletal pathology during the course of this disease in this age group. RESULTS: Average age at time of first presentation was 6 years and 4 months (range 2 wk to 13 yr). Mean follow-up was 6 years and 2 months (range 1-144 mo). Only 2 children required surgery. Two children were diagnosed at birth; 2 were referred as cases of bilateral Legg Calve-Perthes disease. Two children presented with asymmetrical genu valgum; 1 with knee pain, genu valgum, loose bodies, and early degenerative joint disease of both knees; and 2 with limp. CONCLUSION: Although multiple epiphyseal dysplasia is a disease of childhood, it is seldom severe enough to require operative intervention in the initial 2 decades of life. PMID- 15887790 TI - Insulin responses to glucose and isoproterenol decrease with age. AB - BACKGROUND: Older trauma patients are less tolerant of glucose loads than are young patients, in part as a result of diminished insulin response. We hypothesized that diminished insulin responses result from reduced pancreatic sensitivity to beta-adrenergic stimulation. METHODS: We studied healthy subjects prospectively at a clinical investigation unit, to compare young (n = 6, mean age 21 [standard deviation {SD} 2] yr) and older people (n = 6, mean 63 [SD 2] yr). Paired studies of isoproterenol infusion, alone and in conjunction with a 2-hour hyperglycemic glucose clamp, were conducted in each subject. Heart rate, serum insulin concentration and other hemodynamic, biochemical, and physiologic variables were measured. RESULTS: Heart rate increased less markedly in response to isoproterenol in older than in young subjects (p < 0.01). Serum insulin during infusion accompanied by hyperglycemic clamp was also lower (467 [SD 135] pmol/L in older v. 755 [SD 284] pmol/L in young subjects, p < 0.05), despite similar hyperglycemia. Whole-body disposal of exogenous glucose was lower in older (5.8 [SD 1.7] mg/kg/min) than in young subjects (9.6 [SD 3.9], p < 0.05). CONCLUSIONS: Heart rate and serum insulin responses to combined isoproterenol infusion and glucose loading were both lower in healthy older subjects than in the young. Diminished tissue sensitivity to beta-adrenergic signals may contribute to the impaired insulin response and exaggerated glucose intolerance displayed by older patients after injury and during acute illness. PMID- 15887791 TI - An observational study of duplicate presentation rates between two national orthopedic meetings. AB - BACKGROUND: National meetings such as those of the American Academy of Orthopaedic Surgeons (AAOS) and the Canadian Orthopaedic Association (COA) are invaluable in the dissemination of new research findings. Given the limits of meeting agendas, investigators who present the same paper at multiple meetings prevent other presentations on potentially important original research. To determine the incidence of duplicate presentation of research between recent COA and AAOS meetings and between national meetings (AAOS and subspecialty), we conducted an observational study. METHODS: We hand-searched all podium papers and posters from the 2001 COA annual meeting for duplicate presentation at the 2001 and 2002 AAOS annual meetings and subspecialty meetings held in the USA. We evaluated summary data abstracted from the duplicate presentations for consistency. RESULTS: Of 148 presentations at the 2001 COA meeting, 29 presentations (paper and poster) were duplicated at the 2001 or 2002 AAOS meeting: effectively 1 paper in 5 (19.5%). Canadian investigators were significantly more likely to present the same paper at both meetings than Americans (79% v. 13%, respectively; p < 0.01). Those who presented papers at COA altered their AAOS presentations in a variety of ways: by changing the wording in the title of their paper (24% of the time), adding or removing authors (38%), changing authorship order (34%) and changing the sample size (31%). Duplicate presentation rates between AAOS and other orthopedic subspecialty meetings averaged 11.4% (range 3.4%-26.4%). CONCLUSIONS: We identified a 20% duplicate presentation rate between the COA and AAOS annual meetings, and an 11% rate between the AAOS and subspecialty meetings. Stricter enforcement of guidelines and improved dissemination of research findings at both national meetings may limit this practice. PMID- 15887792 TI - Hand-assisted laparoscopic versus open nephrectomies in living donors. AB - Shortages of cadaveric kidneys for transplant into rising numbers of patients with end-stage renal failure have increased the demand for kidneys from live donors. The morbidity associated with traditional open donor nephrectomies (ODN) may discourage many candidates. The newer laparoscopic technique has been promoted as having less morbidity. OBJECTIVES: To evaluate outcomes of hand assisted laparoscopic nephrectomies (HALN) and prospectively compare HALN and ODN. METHODS: After retrospectively reviewing donor and recipient outcomes in 33 HALN (December through August, 2000), we prospectively compared another 47 with 30 ODN performed from September 2000 through April 2001. RESULTS: All 80 HALN were successful, with no requirement to convert to an open procedure. Four donors experienced surgery-related complications: wound infection, retroperitoneal hematoma, prolonged ileus and early small-bowel obstruction, respectively. Two recipients had ureteral complications (1 stricture, 1 leak); 5 experienced delayed graft function, 2 requiring dialysis; and 2 kidneys were lost from infarction. The prospective comparison showed the operative time for HALN (mean 184 min, standard deviation [SD] 39 min) was significantly longer (143 [SD 27] min, p < 0.01), but resulted in less blood loss (p < 0.05). Lengths of time to warm ischemia/early graft function, resumption of oral intake/first bowel movement, and hospital discharge were similar. The abdominal-wall laxity and loss of cutaneous sensation from the flank incision experienced by many ODN patients after was uncommon in the HALN group. Three months after nephrectomy, donor complaints of incisional pain were less common after HALN (p < 0.01). CONCLUSIONS: HALN had good outcomes for donors and recipients, with quicker, more complete recoveries 3 months afterward. PMID- 15887793 TI - Intestinal and peritoneal tuberculosis: changing trends over 10 years and a review of 80 patients. AB - BACKGROUND: To establish the efficiency of minimally invasive procedures in the diagnosis and management of abdominal (intestinal and peritoneal) tuberculosis (AT), I retrospectively and then prospectively evaluated clinical, physical and laboratory findings in patients with AT at a university general-surgery clinic. Diagnostic and therapeutic options were also considered. METHODS: Data from the case records of 43 patients diagnosed with AT via laparotomy were collected and analyzed. On the basis of findings from that early analysis, a further 37 patients were studied prospectively. In the latter group, percutaneous aspiration, endoscopy and laparoscopy were used for diagnosis, except in cases with acute abdominal findings or failure of these diagnostic procedures. Morbidity and mortality rates and mean hospitalization time were evaluated for both groups. RESULTS: In the early group of 43 patients, the morbidity rate was 30%; mortality, 12%; and median hospital stay, 18.3 days (standard deviation [SD] 6.2 d). The diagnosis was confirmed by laparotomy in 18 patients in the prospective group: laparoscopy or endoscopy in 10 and percutaneous drainage of an intra-abdominal abscess in 9. Changes in our approach to diagnostic procedures in the prospective group led to decreased morbidity (11%), mortality (0) and median hospital stay (9.3 [SD 4.7] d). CONCLUSIONS: Minimally invasive procedures such as laparoscopic, endoscopic and percutaneous biopsy should be used for diagnosis of IPTB as a first step in diagnosis. Laparotomy should be performed only when complications develop or diagnosis remains unclear in spite of these diagnostic modalities. PMID- 15887794 TI - Using administrative databases to measure waiting times for patients undergoing major cancer surgery in Ontario, 1993-2000. AB - PURPOSE: To determine how long patients in Ontario waited for major breast, colorectal, lung or prostate cancer surgery in the years 1993-2000. METHODS: "Surgical waiting time" was defined as the interval from date of preoperative surgeon consult to date of hospital admission for surgery. We created patient cohorts by linking appropriate diagnosis and procedure codes from Canadian Institutes of Health Information data. Scrambled unique surgeon identifiers were obtained from Ontario Health Insurance Plan data. Changes in median surgical waiting times were assessed with univariate time-trend analyses and multilevel models. Models were controlled for year of surgery and other patient (age, gender, comorbid conditions, income level, area of residence) and hospital level characteristics (teaching status, procedure volume status). RESULTS: Compared with 1993, median surgical waiting times in the year 2000 increased 36% for patients with breast cancer (to 19 d), 46% with colorectal (to 19 d), 36% with lung (to 34 d) and 4% with prostate cancer (to 83 d). Multilevel models confirmed significant increases in waiting times for all procedures. There were no concerning or consistent differences in waiting times among the categories of hospitals and patients examined. DISCUSSION: There were significant increases in surgical waiting times among patients undergoing breast, colorectal, lung or prostate cancer surgery in Ontario over years 1993-2000. Administrative databases can be used to efficiently measure such waits. PMID- 15887795 TI - Traumatic growth arrest of the distal tibia: a clinical and radiographic review. AB - BACKGROUND: Injury to the physis of the distal tibia in children can lead to subsequent growth arrest. This can result in physeal bars, leg-length discrepancies and angular deformities. METHOD: The cases of 12 children with distal tibial growth arrest due to ankle trauma were reviewed. All were treated at a tertiary care institution between 1990 and 2002. RESULTS: The average age at initial injury was 9.7 years (range 5-13 yr). Salter-Harris classifications (SH) of their injuries were SH2 in 4 children, SH3 in 1, SH4 in 6 and SH5 in 1 child. Four involved open fractures; 5 were high-energy injuries. Six of the injuries resulted in simple physeal bars, 4 caused pure angular deformities and 2 resulted in leg-length discrepancies > 2 cm. Eight of the physeal arrests were treated either with bar excision, selective epiphysiodesis or osteotomy for angular correction. CONCLUSION: This series reinforces the importance of frequent follow up of distal tibial physeal injuries in order to detect growth arrest early, thus facilitating corrective surgery. PMID- 15887796 TI - Users' guide to the surgical literature. Case-control studies in surgical journals. AB - INTRODUCTION: Some articles in surgical journals identify themselves as case control studies, but their methods differ substantially from conventional epidemiologic case-control study (ECC) designs. Most of these studies appear instead to be retrospective cohort studies or comparisons of case series. METHODS: We identified all self-identified "case-control" studies published between 1995 and 2000 in 6 surgical journals, to determine the proportion that were true ECCs and to identify study characteristics associated with being true ECCs. RESULTS: Only 19 out of 55 articles (35%) described true ECCs. More likely to be ECCs were those articles that reported "odds ratios" (ORs) (the OR for being an ECC if a study reported "ORs" compared with those reporting no "ORs" 15.3; 95% confidence interval [CI] 2.8-82.6) and whose methods included logistic regression analysis (OR 3.6, CI 1.0-12.9). Studies that focused on the evaluation of a surgical procedure were less likely to be ECCs (OR 0.2, CI 0.1-0.7) than other types of studies, such as those focusing on risk factors for disease. CONCLUSIONS: The term "case-control study" is frequently misused in the surgical literature. PMID- 15887797 TI - Surgical images: musculoskeletal. Costal hemangioma presenting as rib pain after pneumonia. PMID- 15887798 TI - Surgical Education and Self-Assessment Program (SESAP). Category 1, item 13. PMID- 15887799 TI - Benign granular-cell tumour of the breast. PMID- 15887800 TI - Pure invasive micropapillary carcinoma of the male breast: report of a rare case. PMID- 15887801 TI - Synchondrosis fracture in a pediatric patient. PMID- 15887802 TI - Melanoma erysipeloides. PMID- 15887803 TI - Small-bowel perforation: a consequence of feeding jejunostomy. PMID- 15887804 TI - Implantation metastasis from adenocarcinoma of the colon into a fistula-in-ano: a case report. PMID- 15887805 TI - Patient ethics and evidence-based medicine--the good healthcare citizen. PMID- 15887806 TI - The loyal patient at the end of life: a Roycean argument for assisted suicide. PMID- 15887807 TI - The importance of trust for ethics, law, and public policy. PMID- 15887808 TI - Patients: the Rosetta Stone in the crisis of medicine. PMID- 15887809 TI - Responsibility and health. PMID- 15887810 TI - Special section: patient ethics. PMID- 15887811 TI - Motivation, risk, and benefit in living organ donation: a reply to Aaron Spital. PMID- 15887812 TI - Precaution and solidarity. PMID- 15887813 TI - Everyday disasters. PMID- 15887814 TI - Stem cell science and politics: a talk with Elizabeth Blackburn. PMID- 15887815 TI - Human rights and their role in global bioethics. PMID- 15887816 TI - On personal responsibility and the human right to healthcare. PMID- 15887817 TI - The perils of hope. PMID- 15887818 TI - Fight against depression. PMID- 15887819 TI - Bengali adaptation of brief patient health questionnaire for screening depression at primary care. AB - Depression is a common mental disorder quite prevalent at the primary care level and needs proper identification and prompt treatment. Bengali adaptation and validation of such an instrument (BPHQ) is aimed to provide a sensitive screening instrument that can help primary care physicians in the identification of depression correctly and easily. PMID- 15887820 TI - Bacterial vaginosis and vaginal anaerobes in preterm labour. AB - Maternal genital infections, particularly bacterial vaginosis has been implicated as a cause for preterm labour and adverse pregnancy outcomes. This prospective study aimed to study the association of bacterial vaginosis with preterm labour. The prevalence of bacterial vaginosis was studied in 60 women in preterm labour who had no recognisable cause for prematurity and in 60 term labour controls. Demographic factors, pregnancy outcome and reproductive history were also studied. Vaginal specimens for Gram-stain and culture were collected from posterior vaginal fomix and bacterial vaginosis was defined by evaluation of Gram stained smear by Spiegel criteria. Bacterial vaginosis was diagnosed in 31.6% of women in preterm labour and in 15% of term labour controls ( p<0.05). In preterm labour group, preterm delivery occurred in 48 women (80%) out of which 18 women had bacterial vaginosis and term delivery occurred in only one woman with the condition. Anaerobes were significantly associated with bacterial vaginosis ( p<0.01) and were more common in women with preterm labour ( b>0.05). The results indicates that bacterial vaginosis has a significant association with preterm labour and adverse pregnancy outcome. PMID- 15887821 TI - Comparative evaluation of diabetic and non-diabetic stroke--effect of glycaemia on outcome. AB - Though there are extensive data on diabetic retinopathy, neuropathy and peripheral vascular disease, there are not enough data on diabetic stroke. Present study was carried out to evaluate distinctive feature of diabetic and non diabetic stroke and to compare early mortality between two groups. Four hundred and fifty consecutive stroke cases categorised into group I of 171 known diabetic and 279 non-diabetics. Reactive hyperglycaemia seen in cases with HbA1c below 7.0% were put into group II. Patients were evaluated clinically. Evidence of peripheral vascular disease, retinopathy and other target organ damage were assessed. Glycaemic parameters monitored include fasting plasma glucose (FPG), random plasma glucose (RPG) and HbA1c. Total serum cholesterol was taken as the main risk factor in both groups though lipid profile was done in all cases. CT scan was done in all cases and a repeat one when needed. Out of 450 cases 171 were diabetic and 279 cases were non-diabetic. There was female preponderance in diabetic. Average age of diabetic stroke was lower than non-diabetic, 51.2 years as against 67 years. History of previous stroke was higher in diabetic (15.8%) than non-diabetic (5.7%). There was higher incidence of transient ischaemic attack in diabetic (22.8%) than non-diabetic (7.5%), which was highly significant (p<0.001). Hypertension and hypercholesterolaemia (> 175 mg/dl) were two important risk factors in 70.9% and 30.9% respectively in group I compared to 47.6% and 21.1 % respectively in group II, both of which are significant (p<0.001). Ischaemic stroke were higher in group I(69%) as compared to group II (45.8%) which was significant (p<0.001). Lacunar infarct were more in group I (73.7%) than group II (61.7%). Haemorrhagic stroke was higher in group II (52.7%) than in group I (30.4%). Apart from recent event CT scan showed evidence of old lacunar infarct in 36.8% cases of group I compared to 21.1% of group II. During follow-up mortality within 4 weeks was higher in haemorrhagic stroke of group I (55.8%) compared to 49.6% in group II which is significant (p<0.05). For ischaemic stroke mortality in group I was 26.3% compared to 14.8% in group II which is very significant (p<0.001). Out of total mortality in group I, 35.08% had high HbA1c. Increased risk of stroke in diabetic is probably related to hypertension and lipid abnormalities. Increased mortality from abnormal glycaemic control needs to be evaluated further. PMID- 15887822 TI - Depression and diabetes: a risky comorbidity. AB - Diabetes is a common medical condition and requires careful management. Depression is a prevalent and recurrent condition among both type 1 and type 2 diabetes. Depression complicates diabetes by promoting poor glycaemic control and increasing risk of other complications. Prompt treatment of depression has significant favourable effects on mood and quality of life and beneficial effects on glycaemic control among the diabetic patients. PMID- 15887823 TI - Depression in general clinical practice. AB - Worldwide currently about 340 million people suffer from depression. If not treated, it leads to increased morbidity and mortality. Risk factors include female sex, young age, lower socio-economic study, separated or divorced, positive family history, stressful life events, certain medical illness and so on. Aetiology lies on genetic factors, biochemical abnormality and personality and environmental factors. Clinical features lie on cardinal and other common symptoms. Depression usually starts before 40 years of age, average duration of one episode being 3-9 months. Treatment consists of a variety of psychotherapeutic approaches, pharmacotherapy and electroconvulsive therapy. A clinician may start therapy with low dose SSRIs which is considered as 1st line drugs. To educate patients about anti-depressants is very important. PMID- 15887824 TI - Depression in older people: a point to remember in all specialties. AB - Depression is a common disorder in older people. It is usually undiagnosed in elder patients due to atypical symptoms, masked depressive state, mixed with agitation, psychotic delusions and worsening of physical symptoms already present or multiple pains in extremities. It is a very common associated comorbidity with patients of all disciplines--as in post CVA state or postmyocardial infarction, postoperative state, posthysterectomy or in recovery state of various medical disorders, even in a viral influenza, or in a metabolic medical disorder like diabetes mellitus. Author has studied and analysed 120 patients in different wards of Midnapore Medical College & Hospital and some others usually referred by other physicians at OPD and have found high incidence of depression and a marked improvement of symptoms after a short period of treatment. PMID- 15887825 TI - Key-hole open appendicectomy. AB - Five hundred appendicectomies were done through a small transverse incision designed by the author and it has advantages of both minimal invasive surgery and open surgery along with cost effectiveness. Histopathological picture showed that the maximum number of cases (50%) was of catarrhal type. Retrograde appendicectomy was done in 20% cases. Extension of incision was needed in 4% cases and only 4% had wound infection. All the patients are well after 5 years of follow-up. This method with the incision can be employed by any surgeon and does not require any specialised set-up; it is also very economical. PMID- 15887826 TI - Predictive efficacy of Framingham's risk score in Indian scenario--a retrospective case-control study. AB - To assess the predictive ability of Framingham's risk score in primary prevention in our population, 252 cases and 212 age and sex matched controls were taken up for study. Those patients, who were presenting for the first time with acute coronarysyndrome (ACS) and who did not have any prior manifestations of coronary artery disease (CAD) and whose medical records were available formed the patient group. Framingham's risk score was calculated and the corresponding 10 years risk was assessed in each of them. The patients and controls were divided into two groups--diabetic and non-diabetic. Depending on the 10 years risk, they were further grouped into high risk (10 years risk > 20%), moderately high risk (10 years risk 10 to 20%) and low risk (10 years risk less than 10%). Results were compared and statistically analysed. In the diabetic patients with ACS 14% would have qualified as high risk, 33% as moderately high risk and 53% as low risk whereas in diabetic patients without any manifestation of CAD the distribution was 4% in the high risk, 54% in the moderately high risk and 42% in the low risk. In the non-diabetic subjects, amongst the patients of ACS, 20% would have been in high risk, 39% in moderately high risk and 41% in the low risk. The corresponding figures in the non-diabetic control subjects were 10% in high risk, 22% in the moderately high risk and 68% in the low risk. In the non-diabetic subjects, the mean risk was significantly more in patients than in controls (14.15% versus 8.61%, p <0.01). However, in the diabetic patients there was no significant difference in the mean projected risk between patients with ACS and patients without any manifestation of CAD (11.37% versus 10.41%, p>0.05). PMID- 15887827 TI - Insulin therapy--role beyond glucose control. AB - Larger studies had shown improved patient outcome and lower probability of coronary artery disease in insulin treated groups. The classical lipid abnormalities associated with type 2 diabetes are low HDL-cholesterol concentration and high triglyceride concentration. Insulin usage leads to a decrease in triglyceride concentration, primarily by its effect on the enzyme adipose tissue lipoprotein lipase. Insulin suppresses the enzyme, thereby controlling lipolysis in uncontrolled diabetes. Insulins therapy also improves the endothelial dysfunction especially in people with evident macrovascular complications. Though insulin is noted to increase adrenergic tone and may cause elevation of blood pressure, still patients with insulinoma do not have high blood pressure. Some studies suggest weight gain with insulin therapy, others contradict it. One study suggests that insulin does not affect treatment satisfaction. Insulin is known to improve the glycaemic scenario and also the insulin secretory pattern by reducing the glucotoxicity. PMID- 15887828 TI - A case of renal cell carcinoma in juvenile age group with unusual rim calcification causing diagnostic dilemma. AB - A case of right-sided abdominal lump in a 15-year male was presented. Radiological investigations revealed a solid rounded lesion affecting the upper pole of the right kidney with peripheral rim calcification. After radical nephrectomy, histopathological examination uncovered renal cell carcinoma (clear cell variety) with well capsulated rim calcification. There was no tumour extension/involvement of renal vein and ureter. Postoperative period was uneventful. PMID- 15887829 TI - The current practice of percutaneous transluminal coronary angioplasty (PTCA) and stent deployment and their long-term results in unstable and stable angina. PMID- 15887830 TI - Reviparin sodium clivarine: a review of its therapeutic use. AB - Reviparin sodium (clivarine) is a second generation LMWH, developed with the aim of maximising the antithrombotic action while minimising the risk of haemorrhage. Clivarine has been extensively studied in acute coronary syndrome. Various clinical studies in unstable angina and acute coronary syndrome have proved that clivarine in a dosage of 3436anti-Xa units twice daily is an effective antithrombotic agent. Clivarine has been shown to be as effective as unfractionated heparin (UFH) in thromboprophylaxis and it has less incidence of local haematoma at injection site. At a daily dose of 1432 IU anti-Xa it was found to be as effective as UFH in preventing deep vein thrombosis (DVT) in moderate risk surgery (general and abdominal) and reducing to a significant extent DVT in patients with brace immobilisation of the legs. At a daily dose of 3436 IU anti-Xa reviparin was as effective as UFH or enoxaparin in preventing DVT in high risk orthopaedic surgery and as effective as UFH in prevention of DVT and/or pulmonary embolism (PE) and/or mortality in high risk orthopaedic surgery. In patients with acute venous thrombo-embolism (VTE), reviparin was more effective than UFH in thrombus reduction and at least as effective as UFH in the prevention of clinical recurrence of DVT and/or PE. The use of reviparin is associated with a similar or lower incidence of bleeding complications than UFH. The benefits of reviparin sodium have been demonstrated in a number of clinical trials. PMID- 15887831 TI - An uncommon cause of shoulder pain in acute ischaemic stroke. PMID- 15887832 TI - Criminal negligence in medical practice. PMID- 15887833 TI - Is brain SPECT a suitable modality for evaluation of postradiotherapy posterior fossa brain tumours? A comparative evaluation with contrast enhanced computed tomography. AB - Single photon emission computed tomography (SPECT) of the brain is exquisitely sensitive in detecting viable tumour tissue in the supratentorial region, but its efficacy has not been properly evaluated till date in case of infratentorial posterior fossa tumours. Twenty-four patients with primary posterior fossa brain tumour were included in this study. In each case brain SPECT with 99mTc glucoheptonate (GHA) was performed for the evaluation of disease status. Contrast enhanced computerised tomography of brain was also performed in all the patients. Brain SPECT was positive in four patients with recurrence of tumour as compared to fifteen cases with computed tomography with a mean GHA retention index 5.26 +/ 1.64. Patients with postradiation gliosis (n=9) showed lower GHA retention index of 1.24 +/- 0.27. This study demonstrates that brain SPECT is not sensitive in detecting recurrence of tumour tissue in infratentorial region, as it is in the supratentorial region, with a sensitivity of 20%, accuracy of 45.83% and negative predictive value of 40% and the chance of any single study coming as false negative is about 80%. PMID- 15887834 TI - Guillain-Barre syndrome in children and adolescents--a retrospective analysis. AB - In this study, 55 cases of Guillain-Barre syndrome (GBS) in children and adolescents of 2-18 years of age were analysed retrospectively to study the clinical profile and to evaluate the prognostic value of reduced compound muscle action potential (CMAP) on the need for ventilation and functional outcome. Of the 28 boys and 27 girls 87.3% were bed-bound at peak deficit. Other features were as follows: Bifacial weakness-75%, bulbar weakness-56.4%, need for assisted ventilation-41.8% and albuminocytological dissociation-65.9%. In the ventilated and non-ventilated group no difference was noted in the incidence of reduced CMAP amplitude (p-value > 0.5). At 3 months 83.3% and at 6 months 80.8% cases were ambulant with support. Reduced CMAP amplitude of less than 20% of the lower limit of the normal in at least 2 nerves did not predict the need for ventilation or the chance of independent walking at 3 or 6 months. PMID- 15887835 TI - A study to determine the pharmacokinetics of gatifloxacin following a single oral dose. AB - Gatifloxacin is a broad spectrum fluoroquinolone that offers enhanced Gram positive activity and anaerobic coverage to other fluoroquinolones. The pharmacokinetic parameters (Cmax, AUCo-t, tmax) of this drug have been evaluated to compare the single dose (400mg) bioavailability of gatifloxacin with the reference formulation. High performance liquid chromatography (HPLC) coupled with U-V detector set at 290 nm has been used to determine plasma concentration of 12 human volunteers as per DCGI (Drug Controller General of India) guidelines. The method has been validated over a linear range of 0.25 to 8 microg/ml from plasma. The minimum quantifiable concentration has been set at 0.25 microg/ml (% CV < 10%). The pharmacokinetic parameters are: Cmax = 4.366 +/- 0.44 microg/ml at tmax = 1.83 +/- 0.44 hour, AUCO0-t = 25.26 +/- 2.91 microg hour/ml, AUCo-inf = 33.68 +/- 4.31 microg hour/ml, Kel = 0.094 +/- 0.024/hour and t1/2 = 8.0 +/- 1.92 hour. PMID- 15887836 TI - Role of dynamic external fixator in the management of fractures of distal end of radius. AB - The intra-articular fracture of radius has been an enigma for orthopaedic surgeons since time immemorial. It is a dilemma as to when to use the fixator, when to mobilise after fixator or simply treat it by conservative means. Numerous fixators are in vogue depending on the preference of the surgeon--in the present prospective study the results of dynamic external fixator--which, based on the principle of ligamentotaxis, were evaluated over a 3-year period and the results showed that although excellent radiological results were seen in 90% of the cases yet only 75% of the cases had a excellent functional result. PMID- 15887837 TI - Pneumomediastinum and pneumopericardium--a clinical condition of multispeciality interest. AB - Pneumomediastinum with its usual associations like pneumopericardium has been described in detail including review of literature, aetiopathogenesis, symptoms and signs, complications, necessary investigations and management protocol by the authors in this article of multispeciality interest. PMID- 15887838 TI - Pregnancy complicated by paraplegia due to Pott's spine. AB - Pott's paraplegia associated with pregnancy is a serious problem and is difficult to treat, as not much literature is available regarding its management. Such a case, where the multidisciplinary team approach gave good neonatal and maternal outcome, is reported here. PMID- 15887839 TI - Botryoid rhabdomyosarcoma of vagina--two case reports. AB - Botryoid rhabdomyosarcoma, previously considered a type of embryonal rhabdomyosarcoma, has clinicopathological features distinctive enough to warrant its classification as a separate entity. It almost always occurs in children under 5 years of age. Two cases of botryoid rhabdomyosarcoma are reported here because of their relative rarity. PMID- 15887840 TI - Gastric and intestinal trichobezoar--a case report. AB - Trichobezoars can occur in young females usually with a history of some underlying behaviour disorder. An eighteen-year-old female presented with a fixed right upper quadrant mass. On investigation, a huge mass consisting of hair was found in the stomach, extending into the small intestine. The mass was removed in toto by a standard gastrotomy incision. PMID- 15887841 TI - Subclavian artery thrombosis--management with thrombolysis and angioplasty. AB - A case of acute thrombotic occlusion is presented in a chronically stenosed subclavian artery with subclavian steal phenomenon noted on colour Doppler study and angiography. Because of economic constraints, thrombolysis with angioplasty was done instead of stenting. The patient is on follow-up for the past one year showing considerable improvement. The present case report confirms the efficacy of angioplasty as a suitable option in patients, who cannot afford to have a stent. PMID- 15887842 TI - An unusual presentation of tuberculous lymphadenitis. AB - A 43 years male presented with recurrent epistaxis and had generalised lymphadenopathy on examination. No haematological disorder could be established even after bone marrow aspiration and biopsy but the patient was found to have tuberculosis of the lymph node on histopathology, with severe thrombocytopenia in the peripheral blood and increased platelet precursor in the marrow suggesting peripheral platelet destruction. Anti-tuberculous therapy was started but the patient died due to subarachnoid haemorrhage. PMID- 15887843 TI - Medicolegal aspects of today's practice. PMID- 15887844 TI - Itraconazole: better choice to treat recurrent vulvovaginal candidoses. PMID- 15887845 TI - Assessment of efficacy, safety and tolerability of fixed dose combination of telmisartan 40mg and hydrochlorothiazide 12.5mg in adult Indian patients with mild to moderate hypertension. AB - The objective of the study was to assess the efficacy, safety and tolerability of a fixed dose combination of telmisartan 40 mg and hydrochlorothiazide 12.5 mg in adult Indian patients with mild to moderate hypertension. A prospective, multicentric, open-label, non-comparative, phase IV study was conducted. A total of 353 patients of either sex, between 18- 65 years of age with supine blood pressure (BP) levels of systolic BP (SBP) of 140-200 mmHg and diastolic BP (DBP) of 95-114 mmHg were included. After a placebo run-in period of 2 weeks, each patient received a fixed dose combination of telmisartan/hydrochlorothiazide (40mg/12.5mg) once daily, for 8 weeks. Supine BP was assessed at the end of every 2 weeks. Tolerability and safety were assessed by physical examination, laboratory parameters and evaluation of adverse events. A total of 339 patients completed the study with 14 drop-out cases because of loss to follow-up. There was a significant fall (p<0.05) in both the SBP and DBP starting from the second week as compared to the baseline. Mean SBP had a significant reduction of 23.55 mmHg (15.0%) and 27.79 mmHg (18%) at the end of 6th and 8th week respectively, compared to baseline values. Mean DBP had also had a significant reduction of 12.51 mmHg (12.6%) and 15.17 mmHg (15.3%) at the end of 6th and 8th week respectively, compared to baseline values. This combination was well tolerated with only 3.9% of the total cases reporting mild adverse events like fatigue, dizziness, nausea, diarrhoea etc. The laboratory values were within normal limits. Fixed dose combination of telmisartan/hydrochlorothiazide (40 mg/12.5 mg) once daily has a significant therapeutic effect and a good tolerability profile in adult Indian patients with mild to moderate hypertension. PMID- 15887846 TI - 2005 Winston Health Policy Lecture. PMID- 15887847 TI - Computer skills for the next generation of healthcare executives. AB - Students beginning a career in healthcare administration must possess an array of professional and management skills in addition to a strong fundamental understanding of the field of healthcare administration. Proficient computer skills are a prime example of an essential management tool for healthcare administrators. However, it is unclear which computer skills are absolutely necessary for healthcare administrators and the extent of congruency between the computer skills possessed by new graduates and the needs of senior healthcare professionals. Our objectives in this research are to assess which computer skills are the most important to senior healthcare executives and recent healthcare administration graduates and examine the level of agreement between the two groups. Based on a survey of senior healthcare executives and graduate healthcare administration students, we identify a comprehensive and pragmatic array of computer skills and categorize them into four groups, according to their importance, for making recent health administration graduates valuable in the healthcare administration workplace. Traditional parametric hypothesis tests are used to assess congruency between responses of senior executives and of recent healthcare administration graduates. For each skill, responses of the two groups are averaged to create an overall ranking of the computer skills. Not surprisingly, both groups agreed on the importance of computer skills for recent healthcare administration graduates. In particular, computer skills such as word processing, graphics and presentation, using operating systems, creating and editing databases, spreadsheet analysis, using imported data, e-mail, using electronic bulletin boards, and downloading information were among the highest ranked computer skills necessary for recent graduates. However, there were statistically significant differences in perceptions between senior executives and healthcare administration students as to the extent of computer skills required in areas such as word processing, graphics and presentation, spreadsheet analysis, using imported data, and working with local area networks (LANs). PMID- 15887848 TI - Developing an on-line course in health economics: if I had known then what I know now... AB - Health economics is an important component of professional education in health services administration, public health, and related fields. This article describes the development of an on-line course in health economics for web-based programs in health services administration and pharmacy. The on-line environment offered solutions for some of the problems often encountered in teaching this subject, but raised a set of new issues. The author explains 13 lessons learned in the process of course development and course revision. The course remains a work-in-progress, with some issues, especially in student assessment and course evaluation, yet to be resolved. PMID- 15887849 TI - Politically connected: increasing the political competency of healthcare executives in health policy. AB - In this article we provide a teaching frame of reference for healthcare management students, to more effectively engage in the health policy-making process. We show how future executives can direct or transfer their management skills and abilities to the health policy arena. In doing so, healthcare executives attain a proactive stance in the political process, which is essential to the overall strategic management of healthcare organizations. Using organization theory as a pedagogical tool, we outline the similarities and differences in the management of hospitals versus state legislatures. We identify areas of common ground, and suggest components to frame political advocacy strategy for healthcare executives that strengthen their political competency. Thus, the article provides a teaching framework for health services management students about key intersections between management and policymaking. PMID- 15887850 TI - Next generation leadership: a profile of self-rated competencies among administrative resident and fellows. AB - Healthcare executives and program faculty have voiced concerns that early careerists lack needed competencies for future leadership in the increasingly complex healthcare industry. However, empirical studies of early careerists' competency levels are limited. We sought to describe administrative fellows' and residents' (n = 78, response rate 73.6%) self-rated competency in several key areas and assess how these ratings differed by individuals' gender, age, prior work experience, year of graduate training, and type of degree program. Respondents rated their competence particularly high (41.7% of respondents rated themselves "A") in the domain of interpersonal and emotional intelligence, which included being an effective team leader and member, coaching and developing others, self-awareness, and self-regulation. Lower ratings were in the domains of facilities management and in development and fundraising. Compared to males, females rated their competency in the financial skills domain lower (P-value = 0.04). Age, prior work experience, year of graduate training, and type of degree program were not significantly associated with self-rated competency in any area. These results provide early evidence that may help program faculty and preceptors consider pedagogical approaches that reflect students' vocalized needs and may help to design strategies that effectively cultivate next generation leadership. PMID- 15887851 TI - A civic engagement paradigm for reforming health administration education and recreating the community. AB - The education of students of health administration has traditionally combined both the theoretical and practical to enhance and balance the learning experience. Classroom exposure to the principles of management, law, organizations, and finance is coupled with problem solving, practicum, internship, and administrative residency experiences. However, just as recent years have seen the developmentof courses from managed care and alternative delivery systems to total quality management and continuous quality improvement, there is also emerging an awareness of the need to enhance the practical side of the learning equation. Perhaps this need is finding expression in curricular opportunities for students to learn from a participatory model known as civic engagement (CE). CE is a way of integrating academic study and community service to strengthen learning while promoting civic and personal responsibility to strengthen communities. Based on experiences with graduate and undergraduate students spanning the last ten years at Texas State University--San Marcos (Texas State), it is suggested that a CE paradigm has been developed within the Department of Health Administration that merits consideration by other programs of health administration. As a model for change, it has the potential for reforming both health administration education and most other higher education disciplines as well. PMID- 15887852 TI - Gender differences in career progression and career satisfaction among graduates of a midwestern M.H.S.A. program. AB - This article compares factors influencing career success among male and female health services manager alumni. A sample of 833 M.H.S.A. graduates received a mail out 27 item questionnaire. Response rate was 48 percent. Factor analysis using Varimax rotation indicated three variables defined as effort, environment and perception, each accounting for 18.18 percent, 16.23 percent, and 10.95 percent of the variance respectively. Independent sample t-tests comparing male versus female scores for factors effort, environment, and perception indicated no statistically significant difference for effort; however there were statistically significant differences for environment and perception. Using a list-wise selection procedure, a sub-sample of 166 cases was further analyzed. Factor scores for effort, environment, and perception were calculated and entered into a regression model to predict career satisfaction. All three factors entered the model at a significance level .05 or less. The authors indicate that because males and females see the influence of these factors differently, there are implications for academic programs and the profession. Academic programs need to discuss concerns about the environment and perceptions and their effect on career progression. Likewise leaders of healthcare organizations should take note of the role that workplace environment and perceptions have in one's career progression. PMID- 15887853 TI - Dehydroepiandrosterone sulfate and bone resorption rates as reflected by serum levels of C-terminal telopeptide of type I collagen: a study in healthy men. AB - Sex steroid hormones contribute to the physiological regulation of bone turnover in males. To address this issue, we investigated serum estradiol (E2), total testosterone (T), and DHEAS concentrations, along with serum levels of carboxy terminal telopeptide of type I collagen (sCTx), in a sample of 76 healthy men aged 23 to 87. The concentration of sCTx declined with age. Both T and DHEAS, at variance with E2, showed a significant age-related decline. T, DHEAS and sCTx significantly (p<0.01) correlated with each other. DHEAS and T were significantly associated after correcting for age (r=0.35, p=0.002) or body mass index (r=0.65, p<0.0001). DHEAS, but not T, significantly correlated with sCTx after correcting for age (r=0.26, p=0.026, and r=0.20, p=0.08, respectively). Stepwise multiple regression analysis showed that only DHEAS (but not T or E2) was a significant independent predictor of sCTx (p=0.0001). Our results show that adrenal androgens play a crucial role in regulating bone resorption in aging men. PMID- 15887854 TI - Human galectin-3 immunoexpression in thyroid follicular adenomas with cell atypia. AB - Human galectin-3 (hgal-3) is a beta-galactoside binding protein involved in a number of physiological and pathological processes. Increasing hgal-3 immunoexpression has been reported in several human tumors, including thyroid carcinomas, but not in benign thyroid lesions. We analyzed the immunolocalization of hgal-3 in cell compartments of benign and malignant thyroid lesions. Hgal-3 immunoperoxidase reaction was carried out on 133 thyroid tissue samples obtained from 113 patients; 20 of these were normal (NT), 85 were benign thyroid lesions [20 colloid nodules (CN), 21 nodular hyperplasias (NH), 7 focal lymphocytic thyroiditis (FLT), 15 Hashimoto's thyroiditis (HT), 22 follicular adenomas (FA)], 25 differentiated carcinomas [15 papillary carcinomas (PC), 6 follicular carcinomas (FC) and 4 Hurthle cell carcinomas (HC)] and 3 anaplastic carcinomas (AC). Among the malignant thyroid lesions, hgal-3 was detected in 12/15 (80%) PC, 3/4 (75%) HC and in 4/6 (66.6%) FC, but in none of the 3 AC. Conversely, hgal-3 immunoexpression was absent in NT and in all benign thyroid lesions, but 1/15 HT and 10/22 (45.4%) FA. In the latter, hgal-3 was mostly expressed in microfollicular areas and in five of the six atypical FA. Hgal-3 cytoplasmic perinuclear immunolocalization was observed in the majority of thyroid carcinomas and in more than half of the FA, theoretically suggesting an involvement of this protein in thyroid tumorigenesis throughout an antiapoptotic activity. Moreover, hgal-3 expression in FA might anticipate the likelihood of evolution of these benign lesions towards malignancy. PMID- 15887855 TI - Growth hormone secretion in primary and secondary hyperparathyroidism. AB - Primary hyperparathyroidism (PHP) is associated with impaired GH secretion. Whether this effect is due to hypercalcemia or to increased serum PTH concentration is unclear. However, patients with familial hypocalciuric hypercalcemia (FHH), who have normal PTH and increased serum calcium concentrations, also have an impaired GH secretion, suggesting that calcium rather than PTH is responsable for this effect on GH secretion. To further investigate this issue, 10 consecutive patients with secondary hyperparathyroidism (SHP) due to vitamin D deficiency were evaluated by the GH response to GHRH+arginine (Arg) test. A group of 60 consecutive untreated PHP patients served as controls. Mean GH response to GHRH+Arg test was 15.8+/-14 microg/l and 37.5+/-16 microg/l (p<0.001) in PHP and in SHP patients, respectively. Forty-two out of 60 (70%) PHP patients had a suppressed or blunted GH response, whereas all SHP patients had normal GH response. The results of the present study confirm and extend our previous observations that PHP is associated with an impaired GH secretion in the majority of cases, and indicate that SHP patients have no abnormality of GH secretion. Thus, hypercalcemia rather than increased serum PTH is responsible for the abnormality of GH secretion. PMID- 15887856 TI - Association between vitamin D receptor FokI. Polymorphism and serum parathyroid hormone level in patients with chronic renal failure. AB - We investigated the relationship between vitamin D receptor (VDR) start codon polymorphism and serum levels of PTH, calcidiol, and calcium in 64 Spanish patients with chronic renal failure (CRF). An exon 2 fragment of the VDR gene was amplified by PCR, and cleaved with the restriction enzyme FokI. The alleles were identified according to the digestion pattern obtained as F (absence of restriction site) and f (presence of restriction site). Genotype frequencies in the patient population were 54.7% FF, 28.1% Ff and 17.2% ff, vs 46.7% FF, 43.3% Ff and 10% ff in a healthy control population. The difference between the two populations was statistically significant (p<0.01). Within the patient population, mean serum PTH level in the FF group was significantly higher (159.77+/-25.69 pg/ml) than in both the Ff and ff groups (106.67+/-19.07 and 77.55+/-15.85 pg/ml, respectively; p<0.05). However there were no significant differences in serum levels of calcidiol or calcium among genotypes. These results suggest that FokI polymorphisms of the VDR gene may determine parathyroid response in CRF patients. PMID- 15887857 TI - Risk factors associated to kidney stones in primary hyperparathyroidism. AB - Nephrolithiasis is the most important clinical manifestation of primary hyperparathyroidism (PHPT), although nowadays this disorder is often asymptomatic. Clinical or biochemical differences between PHPT patients with and without nephrolithiasis have not been clearly identified in most of the previous studies. The aim of the study was to investigate clinical and biochemical parameters in kidney stone former (SF) and non-stone former (NSF) patients with PHPT in order to identify potential risk factors. Serum and plasma samples from 55 consecutive patients (43 females, 12 males) with PHPT were collected after overnight fasting; 24-h urine collection and a fresh sample of urine for sediment analysis were obtained from all patients. Clinical data were recorded in all. Out of 55 patients, 22 had kidney stones, which were symptomatic in 73%. SFs showed circulating PTH, total and ionized calcium, 1,25 dihydroxyvitamin D3, urinary calcium excretion and 24-h urine oxalate levels significantly higher than NSFs. Hypercalciuria was often concomitant with massive quantities of calcium oxalate crystals in urine sediment. Hypercalciuria and relatively high oxaluria were associated with stone formation with an odds ratio (OR) of 4.0 and 7.0, respectively, which rose to 33.5 when they coexisted. Hypomagnesuria and hypocitraturia were common in at least one third of all PHPT patients, but they were not associated to an increased OR. As expected, they were positively correlated with urine calcium excretion, suggesting that calcium, magnesium and citrate are commonly regulated at renal level. In conclusion, hypercalciuria, higher oxalate excretion and severe PHPT are associated with kidney stones in PHPT. PMID- 15887858 TI - Effect of the peroxisome proliferator-activated receptor-gamma C161T polymorphism on lipid profile in Brazilian patients with Type 2 diabetes mellitus. AB - AIM: The aim of the present study was to examine the effects of the C161T polymorphism of the peroxisome proliferator-activated receptor gamma (PPARgamma) gene in Brazilian subjects with Type 2 diabetes mellitus (T2DM) and controls residing in Sao Paulo City, Brazil. METHODS: Genomic DNA was obtained from 207 patients with T2DM and 170 unrelated normoglycemic individuals (CG). Anthropometric data included: body mass index, waist, hip, waist-to-hip ratio; biochemical parameters: fasting plasma glucose, total cholesterol, HDL- and LDL cholesterol, triglycerides, glycated hemoglobin and insulin. Systolic and diastolic blood pressure was also measured. Screening for mutations in the entire coding region of the PPARgamma gene was carried out by PCR, single strand conformational polymorphism analysis (SSCP) and sequencing. C161T polymorphism was analyzed by PCR-RFLP. RESULTS: The C161T polymorphism was the only variant found in exon 6 of the PPARgamma gene. The frequency of the 161T allele in T2DM (0.10) was similar to that found in CG (0.07, p=0.210). Serum triglycerides (p=0.040), VLDL-cholesterol (p=0.040) and Atherogenic Index of Plasma (AIP; p=0.003) were significantly lower in 161T allele carriers than non-carriers in women of the T2DM group. CONCLUSIONS: Our results show that the C161T polymorphism in the PPARgamma gene is not associated with variables related to T2DM or insulin resistance in the Brazilian population. However, a reduction of serum triglycerides and AIP was observed in women with 161T allele of the C161T polymorphism of the PPARgamma gene. PMID- 15887859 TI - Proteomic analysis of human thyroid cell lines reveals reduced nuclear localization of Mn-SOD in poorly differentiated thyroid cancer cells. AB - Differential protein arrays between nuclear extracts of human thyroid cell lines obtained from tumors with different degree of differentiation were exploited to define molecular alterations occurring during thyroid tumor progression. Nuclear extracts from the well differentiated TPC-1 (from papillary carcinoma) and the poorly differentiated ARO (from anaplastic carcinoma) cells showed an overall similar pattern of protein expression as revealed by two-dimensional gel electrophoresis analysis. However, manganese-superoxide dismutase (Mn-SOD) was clearly identified by mass spectrometry procedures as significantly less expressed in ARO compared to TPC-1 cells. A reduced expression of Mn-SOD in the nuclear compartment was confirmed by Western blot and immunofluorescence analysis. A similar expression pattern of nuclear Mn-SOD was detected by immunohistochemistry in human thyroid tumors, with the lowest or absent detection in anaplastic carcinomas. Moreover, the levels of nuclear Mn-SOD in tumor cells were lower than in the normal thyrocytes. These data indicate that an altered nuclear expression of Mn-SOD parallels, together with changes in other elements of the antioxidant protective system, the loss of differentiation occurring during the progression of thyroid tumors. PMID- 15887860 TI - Is serum C-reactive protein concentration correlated with HbA1c and insulin resistance in Type 2 diabetic men with or without coronary heart disease? AB - BACKGROUND AND AIMS: C-reactive protein (CRP) is an inflammatory marker that predicts coronary heart disease (CHD) risk. Diabetes mellitus (DM) counts as a CHD risk equivalent. We aimed to compare serum high sensitivity CRP (hs-CRP) levels in Type 2 diabetic (T2DM) men without CHD, non-diabetic CHD patients and T2DM patients with CHD. SUBJECTS AND METHODS: Four groups were formed; Group 1 [DM(+), CHD(-), no.=25], Group 2 [DM(-), CHD(+) no.=25], Group 3 [DM(+), CHD(+), no.=25], and Group 4 (controls, no.=30). Serum hs-CRP, insulin, glucose, total, HDL-, LDL- and VLDL-cholesterol, triglyceride levels and homeostasis model assessment for insulin resistance (HOMA-IR) index were determined. RESULTS: Mean hs-CRP level of Group 1 (0.6+/-0.29) was not different statistically from Group 2 (1.44+/-0.97). Mean hs-CRP levels were higher in men with CHD, whether they were diabetic (Group 3; 3.83+/-2.01 mg/dl) or non-diabetic (Group 4), than in control subjects (0.16+/-0.15; p=0.0001 and p<0.004, respectively). Mean hs-CRP level of Group 3 was also higher than Group 2 (p=0.0001). There was a positive correlation between serum hs-CRP and glycated hemoglobin (HbA1c; r=0.277, p<0.01), fasting insulin (r=0.336, p<0.02) and HOMA-IR (r=0.348, p<0.02) in T2DM men with or without CHD. CONCLUSIONS: T2DM men without CHD had similar CRP levels with non diabetic CHD patients, whereas CRP levels of T2DM men with CHD were higher than non-diabetic men with CHD. Because of a positive correlation between serum hs-CRP and HbA1c, fasting insulin and HOMA-IR, inflammation, insulin resistance and hyperglycemia jointly contribute to the cardiovascular risk in T2DM men. PMID- 15887861 TI - Symptomatic hyponatremia as a presenting sign of hypothalamic-pituitary disease: a syndrome of inappropriate secretion of antidiuretic hormone (SIADH)-like glucocorticosteroid responsive condition. AB - Hyponatremia associated with high urine osmolality is usually caused by inappropriate secretion of antidiuretic hormone. However, secondary hypoadrenalism is also accompanied by hyponatremia and with features indistinguishable from the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). As secondary hypoadrenalism requires a specific treatment, a high index of suspicion and appropriate hormonal testing are required to differentiate between these two entities. We retrospectively studied 10 patients with a previously undiagnosed hypothalamic-pituitary disease who presented with an acute symptomatic hyponatremia. Mean age (+/-SD) was 65.1+/-8.4 yr. Mean serum sodium was 120.7+/-2.9 nmol/l and urinary osmolality, 453.9+/-74 mosmol/kg. Serum creatinine, urea and uric acid were low. Mean serum morning cortisol was low, 104.0+/-55.2 nmol/l. High-dose ACTH test showed adequate increment of serum cortisol in 3 out of 7 patients tested. Two of these 3 patients did not respond adequately to the low-dose ACTH test. Endocrine evaluation disclosed partial or complete hypopituitarism in all 10 patients. Six patients had pituitary macroadenomas, one had a craniopharyngioma, one patient had a large aneurysm of the internal carotid with sellar destruction and two others had empty sella. Treatment by fluid restriction did not affect serum sodium levels significantly. In contrast, all patients achieved normal sodium when treated by glucocorticosteroid. Central hypoadrenalism should be considered in any patient presenting with hyponatremia with high urine osmolality. Low-dose ACTH test should be performed and followed by appropriate endocrine and imaging studies. Hyponatremia in these patients is promptly corrected by glucocorticosteroid replacement. PMID- 15887862 TI - Characteristics of adult patients with growth hormone deficiency who underwent neurosurgery for functioning and non-functioning pituitary adenomas and craniopharyngiomas. AB - The aim of the present study was to evaluate the characteristics of GH deficiency (GHD) in adult patients after neurosurgery for pituitary adenomas and craniopharingiomas. One hundred and one GHD patients, (42 F/59 M), aged 47.58+/ 14.4 yr (mean+/-SD; range 21-78), body mass index (BMI) 28.6+/-0.6, with a history of adult-onset hypothalamic-pituitary disease, were recruited for the study. The whole group included: 45 non-functioning pituitary adenomas, 23 craniopharyngiomas, 16 PRLomas, 8 GHomas, 7 ACTHomas and 2 FSHomas; in particular 51 were macroadenomas and 27 microadenomas. At study entry, GHD diagnosis was carried out by assessing GH secretion after GHRH+arginine. All patients were submitted to the study at least 12 months after neurosurgery and, where needed, subjects were replaced with an appropriate treatment. GHD was mild in 3/101 (3%) and severe in 98/101 patients (97%). Other hormone deficiencies associated with GHD were considered: TSH, ACTH, FSH/LH, ADH. The distribution of peak GH among all patients, according to the type of disease before neurosurgery, showed that patients with Cushing disease were characterized by the presence of higher peak GH. According to the number of additional hormone deficits, the distribution of peak GH among all patients was as follows: GHD was isolated in 4/101 subjects (4%; group A), while it was associated with 1 (14/101, 14%; group B), 2 (22/101, 22%; group C), 3 (44/101, 43%; group D) and 4 hormone deficits (17/101, 16%; group E). GHD was severe in all patients in the panhypopituitaric group. Total IGF-I plasma levels in the whole group of GHD patients were 95.2+/-4.2 microg/l. In all groups of patients IGF-I was lower in subjects with severe GHD than in those with mild GHD (93.6+/-4.1 vs 148.6+/-33.6 microg/l, p<0.03). In particular, according to the type of disease presented before neurosurgery, patients with Cushing disease were characterized by the presence of higher IGF-I plasma levels compared to the other. According to the number of additional deficits, the distribution of IGF-I plasma levels was characterized by higher values when GHD was isolated than when it was associated with multiple hormone deficiencies. IGF I plasma levels were positively associated to peak GH during GHRH+arginine (r=0.4, p<0.0005). We conclude that patients after neurosurgery approach for sellar and parasellar neoplasia, within an appropriate clinical context, and both the presence of additional pituitary hormone deficiency and low levels of IGF-I can be considered a clear GHD condition, and therefore do not require provocative tests evaluating GH secretion. PMID- 15887863 TI - Acromegaly due to a growth hormone-releasing hormone-secreting intracranial gangliocytoma. AB - In more than 95% of cases acromegaly is due to GH hypersecretion by a pituitary adenoma. GHRH hypersecretion accounts for about 0.5% of cases of acromegaly. Intracranial GHRH-secreting tumors are extremely rare and only a few well documented cases have been reported. The clinical features of acromegaly due to intracranial GHRH-secreting tumor are indistinguishable from those of other patients with "classical acromegaly". In cases of intrasellar gangliocytomas, not even radiological findings help to make the correct diagnosis, which can only be made with the hystological study. We present the case of a woman with acromegaly; the magnetic resonance demonstrated a 2x1.8x1.2 cm mass in the jugum sphenoidalis region, associated with a partial empty sella. There was a partial response to high-dose lanreotide therapy, so surgical treatment was decided, although only part of the tumor could be removed. Histopathological diagnosis was consistent with gangliocytoma, and immunostaining in the ganglionic cells was positive for GHRH. After surgery, hormone hypersecretion persisted, so medical treatment was reintroduced. In summary, we report a well-documented case of an intracranial GHRH-secreting gangliocytoma, an exceedingly rare cause of acromegaly. Clinical and biochemical data did not allow to make the correct diagnosis, which was only made on the pathological study. This case underscores that acromegaly can be due to causes other than a GH-secreting adenoma, and underlines that finding an image not typical of a pituitary adenoma should raise the suspicion that an unusual cause subsides the acromegaly. PMID- 15887864 TI - Pituitary tumor disappearance in a patient with newly diagnosed acromegaly primarily treated with octreotide LAR. AB - We describe the case of an acromegalic patient primarily treated with octreotide LAR in whom the pituitary tumor disappeared after 18 months of treatment. A 68-yr old woman, with clinical suspicion of acromegaly, was admitted to our Unit with the ultrasonographical evidence of cardiac hypertrophy, arrhythmias, right branch block and interatrial septum aneurism. She referred hands and feet enlargement since the age of 30 and facial disfigurements since the age of 50. At the age of 45 she underwent surgery for carpal tunnel syndrome and at the age of 61 an euthyroid nodular goiter was diagnosed. Hormonal evaluation showed elevated circulating GH levels (25+/-3.2 ng/ml), not suppressible after oral glucose load, and elevated IGF-I levels (646 ng/ml), whereas the remaining pituitary function was normal. Visual perimetry was normal, whereas magnetic resonance imaging (MRI) showed an intrasellar pituitary adenoma with maximal diameter of 9 mm. In order to improve cardiovascular function before surgery, the patient started octreotide LAR 20 mg every 4 weeks for 3 months. Then based on IGF-I values, the dose was adjusted to 30 mg. After 6 months a second MRI showed significant tumor reduction (>50% of baseline maximal diameter), GH and IGF-I were within the normal range and the patient continued the treatment. After one-year therapy, an improvement of cardiac alterations was recorded and the patient was referred to the neurosurgeon. However, she refused the operation. At 18-month follow-up, MRI showed the complete disappearance of direct and indirect signs of pituitary adenoma. To our knowledge, this is the first case of complete radiological remission of pituitary tumor during octreotide LAR treatment in acromegaly. PMID- 15887865 TI - Diagnostic difficulties in long-standing insulinoma with near-normal plasma insulin levels. AB - Insulinoma tumors are often difficult to detect, as the symptoms largely precede occurrence of a visualized tumor. We report the case of an insulinoma patient with long delayed diagnosis and marked adaptation to extreme hypoglycemia. The patient with a 7-yr history of seizures was found to have plasma glucose concentration during a starvation test as low as 16 mg/dl, with no clinically significant symptoms and concomitant normal plasma insulin levels: 10-30 microIU/ml. All attempts to localize a tumor with repeated abdominal ultrasound examinations or computed tomography scanning were unsuccessful. The patient did not tolerate the introduced oral treatment with diazoxide. Once it had become technically available, endoscopic ultrasonography of the pancreas was performed. It revealed a 10 mm tumor in the pancreatic head. The tumor was subsequently removed surgically. During the operation plasma insulin concentration rose almost 15-fold, which confirmed the insulin-secreting character of the growth. Microscopic examination revealed benign insulinoma, with partially trabecular structure. One month after the operation the patient had normal plasma glucose values of 60-120 mg/dl, but she constantly complained of excessive thirst, which occurred soon after the operation and slowly subsided in the following weeks. In conclusion, the present report demonstrates that insulinoma should be considered and searched for in every case of hypoglycemia associated with normal insulin levels. It also confirms the essential role of endoscopic ultrasonography in the diagnosis of insulin-secreting tumors. PMID- 15887866 TI - Intrahepatic cholestasis as a paraneoplastic syndrome associated with pheochromocytoma. AB - Pheochromocytoma is a rare tumor of chromaffin cells that secrete catecholamines and several cytokines. The clinical manifestations are protean and may include hypertension, weight loss, sweating, palpitation, headache, anxiety, tremor, nausea, vomiting, and hypercalcemia. The tumor can mimic many unrelated diseases, leading to significant delay and difficulty in diagnosis. We report a case of a 37-yr-old male admitted with jaundice, dark urine, fever, and signs of a systemic inflammatory response. Abdominal computed tomography revealed a heterogeneously enhancing tumor between the pancreatic tail and left kidney. There was no evidence of obstruction to bile flow, neoplastic involvement of the liver or bile ducts, or infectious etiology. The tumor was removed and found to be a pheochromocytoma. Immunohistochemical analysis revealed the presence of interleukin-1beta in the tumor cells. After surgery, the jaundice resolved without further treatment, leading us to the conclusion that it was a paraneoplastic phenomenon possibly related to interleukin-1beta production. We suggest that occult pheochromocytoma should be added to the differential diagnosis of unexplained intrahepatic cholestasis. PMID- 15887867 TI - Serum 25-hydroxyvitamin D and parathyroid hormone exhibit threshold behavior. PMID- 15887868 TI - There is no lower threshold level for parathyroid hormone as 25-hydroxyvitamin D concentrations increase. PMID- 15887869 TI - 21-Hydroxylase deficiency: from molecular genetics to clinical presentation. AB - Congenital adrenal hyperplasia due to deficiency of the enzyme 21-hydroxylase (21 OH), a cytochrome P450 enzyme located in the endoplasmic reticulum and which catalyzes the conversion of 17-hydroxyprogesterone to 11-deoxycortisol and progestene to deoxycorticosterone, is distinguished in its classical and non classical form and is also one of the most common autosomal recessive inherited diseases in humans. The classical form appears in a rate between 1:5000 and 1:15,000 among the live neonates of North America and Europe, while the non classical form occurs in approximately 0.2% of the general white population. This rate is especially high between the Ahskenazi Jews and a part (ie Italians, Hispanics) of the Mediterranean populations. Three alleles are associated with the 21-OH locus and can be combined in several ways in individuals who are either unaffected, heterozygote carriers, or affected with classical or non-classical disease. Variable signs and symptoms of hyperandrogenism, such as hirsutism, acne, virilization of the external genitalia and/or the body, short stature, menstrual irregularities, are common to both types of the disorder. Among the genes responsible for the synthesis of the enzyme 21-OH and the antigens of HLA system, exist both a proven genetic linkage and a proven genetic linkage disequilibrium. HLA-Bw47, HLAB5 and HLA-B35 are the most common haplotypes usually met in the classical form, while the haplotype HLA-B14DR1 is the most recurrent in the non-classical form of the disease. The significant advances in molecular biology and gene analysis over the past two decades have led to the development of novel sensitive methods of DNA analysis and study, such as polymerase chain reaction and southern blot analysis. Thus, it has been revealed that the synthesis of enzyme 21-OH is controlled by two genes, the active CYP21B gene and the CYP21A pseudogene. All three forms of the disease have a known sequence of gene changes owing to mutations in isolated proteins or whole series of genes due to translocations or deletions of genetic material. PMID- 15887870 TI - Growth hormone treatment in non-growth hormone-deficient short children. AB - The unlimited availability of GH obtained by recombinant DNA technology has allowed optimization of treatment in GH-deficient (GHD) children. At the same time it has prompted a number of studies in conditions not characterized by GHD such as Turner syndrome, intrauterine growth retardation, chronic renal failure and other chromosomal and genetic abnormalities associated with short stature. Several controlled and uncontrolled studies have now reported the adult height of patients with short stature and normal GH secretion. Critical reviewing of the data shows that some short non-GHD children may benefit from a prolonged treatment with GH. However, further studies are needed in order to be able to identify the subjects for whom treatment is really beneficial. PMID- 15887871 TI - "La Monstrua Vestida", a case of Prader-Willi syndrome. PMID- 15887872 TI - A shortened psychophysical task to quantify irritability: the Reactive Irritability Scale II (RIS-II). AB - The Reactive Irritability Scale (RIS) uses magnitude estimation to measure reactive irritability. Respondents rate target sounds in comparison to a neutral reference sound. The RIS proved more sensitive than self-report measures to detect irritability associated with withdrawal from cigarette smoking and with exposure to a stressful environment, but was too long (30 min) for routine use. We report here on a shortened version (13 min)--RIS-II. The RIS-II exhibited robust test-retest reliability and correlated strongly with the original RIS (Study 1). In Study 2, the RIS-II was administered to subjects who experienced psychological stress and then were exposed to progressive relaxation, music, cognitive tasks, or silence; the Progressive Relaxation group was the most irritable. In Study 3, the RIS-II was administered to chronic pain patients. Pain patients found the sounds less irritating than did controls with the exception of the reference sound; repeated presentation of the reference sound increased irritability. These studies indicate that the RIS-II is a reliable instrument that may have utility for the measurement of irritability in laboratory and clinical settings. In addition, these studies indicate that the RIS-II is understandable by individuals of different ages who are from educationally- and culturally-diverse backgrounds and individuals who are healthy as well as individuals suffering from chronic medical conditions who are on multiple medications. PMID- 15887873 TI - Health events and the smoking cessation of middle aged Americans. AB - This study investigates the effect of serious health events including new diagnoses of heart attacks, strokes, cancers, chronic lung disease, chronic heart failure, diabetes, and heart disease on future smoking status up to 6 years postevent. Data come from the Health and Retirement Study, a nationally representative longitudinal survey of Americans aged 51-61 in 1991, followed every 2 years from 1992 to 1998. Smoking status is evaluated at each of three follow-ups, (1994, 1996, and 1998) as a function of health events between each of the four waves. Acute and chronic health events are associated with much lower likelihood of smoking both in the wave immediately following the event and up to 6 years later. However, future events do not retrospectively predict past cessation. In sum, serious health events have substantial impacts on cessation rates of older smokers. Notably, these effects persist for as much as 6 years after a health event. PMID- 15887874 TI - The validity of Beck Depression Inventory and Hamilton Rating Scale for Depression items in the assessment of depression among patients with multiple sclerosis. AB - Measuring depression in patients with chronic illnesses such as multiple sclerosis (MS) is potentially complicated by the fact that several somatic symptoms of depression are also common in chronic illnesses. Whether standard assessment measures such as the Beck Depression Inventory (BDI) and Hamilton Rating Scale for Depression (HRSD) should exclude certain somatic symptoms when used in MS has been examined previously, but there is no clear consensus on this issue. The present study evaluated the utility of individual BDI and HRSD items for assessing depression in MS patients by examining how individual items responded to depression treatment in 42 (29 female) depressed MS patients. All 21 BDI items and 12 of 17 HRSD items decreased significantly with treatment, suggesting that all BDI items tap depression, as do 12 of 17 HRSD items. Thus, the present data support the inclusion of all BDI items when measuring depression in MS. Decisions on whether or not to use all HRSD items or only the 12 shown here to capture depression may depend on the study purpose and design. PMID- 15887875 TI - Patient strategies to cope with high prescription medication costs: who is cutting back on necessities, increasing debt, or underusing medications? AB - Many chronically ill adults in the United States face high prescription medication costs, yet little is known about the strategies patients adopt to cope with these costs. Through a national survey of 4,055 adults taking prescription medications for one of five chronic diseases, we compared whether respondents cut back on necessities such as food or heat to pay for medications, increased debt, or underused medications because of cost. We also examined the sociodemographic and clinical correlates and differential use by different sub-groups of these three strategies. Overall, 31% of respondents reported pursuing at least one of the strategies over the prior 12 months. Twenty-two percent had cut back on necessities, 16% had increased their debt burden, and 18% had underused prescription drugs. Among patients who underused their medication, 67% also had cut necessities or increased debt. Although we found significant differences in the way patients with varying socio-demographic characteristics responded to medication cost pressures, use of all these strategies was especially common among patients who were low-income, in poor health, and taking multiple medications. PMID- 15887876 TI - Mediation and modification of the association between hopelessness, hostility, and progression of carotid atherosclerosis. AB - Hopelessness and hostility are linked to progression of carotid atherosclerosis (PCA). The purpose of this study was to replicate such relations and to evaluate the role of biological pathways involving hyperactivation of the sympathetic nervous system (SNS) and the hypothalamic-pituitary-adrenocortical (HPA) axis. PCA was evaluated by 4-year change in three ultrasound measures of intima-media thickness (IMT) in 1027 men aged 42-60 years at baseline. Effect modification and mediation of relationships between psychosocial factors and PCA were examined for the measures systolic blood pressure (SBP), fibrinogen, and waist-to-hip ratio (WHR), levels of which are indicative of activity along these pathways. Hopelessness and hostility were associated with PCA. Fibrinogen mediated to a moderate extent the association between hopelessness and PCA. SBP significantly modified the relation between hostility and PCA in participants of moderate hostility. The above biological pathways are implicated in the mechanisms connecting hopelessness, hostility, and PCA. PMID- 15887877 TI - Distress associated with prenatal screening for fetal abnormality. AB - A theoretically-based, multivariate approach was used to identify factors associated with emotional distress for pregnant women undergoing maternal serum alpha fetoprotein (MSAFP or AFP) testing, used to detect abnormalities of the fetal brain and spinal cord. Participants were those who received normal results (N = 87). Study results supported the hypothesis that different factors would predict distress before and after testing. Satisfaction with information about testing predicted lower emotional distress early in the testing process; concerns about the child having other medical conditions and low-dispositional optimism predicted distress later. Study findings indicate that even in women who receive normal test results, AFP testing is associated with a modest degree of emotional disturbance which declines, but does not completely abate, after testing. PMID- 15887878 TI - Psychosocial characteristics associated with sun protection practices among parents of young children. AB - Despite the importance of sun protection during childhood, many parents fail to adequately protect their children from the sun. The current study assessed sun related knowledge, stage of change, and psychosocial characteristics of parents of young children. We examined how knowledge and psychosocial characteristics differed by stage of change, and tested a model of the relationship among demographics, knowledge, psychosocial variables, and child sun protection. Surveys were completed by 391 parents of preschool-aged children. Parent knowledge about skin cancer was relatively high and did not vary by stage of change. Parents in earlier stages of change perceived more barriers to child sun protection, had more positive beliefs about children's sun exposure, perceived their child to be less susceptible to sun damage, and had lower self-efficacy for child sun protection than did parents in later stages of change. In a path model, the strongest predictors of child sun protection were parents' own sun protection habits, perceived barriers, and self-efficacy. Results highlight the importance of parent psychosocial characteristics in predicting child sun protection. PMID- 15887879 TI - Benefit-finding among patients with rheumatoid arthritis: positive effects on interpersonal relationships. AB - This longitudinal study of patients with rheumatoid arthritis used mixed methods to identify and describe the positive effects of illness on relationships, examine correlates of benefit-finding, and test the relationship between benefit finding and adjustment outcomes. When asked about interpersonal benefits of their illness, 71.3% of the respondents described interpersonal benefits, whereas 16.2% reported another type of benefit, and 12.5% reported no benefits. The most frequently described benefit was appreciation of support received from loved ones. Less pain, lower psychological distress, and perceiving fewer social constraints were related to finding interpersonal benefits in the illness experience. Interpersonal benefit-finding predicted lower levels of disability at a 12-month follow-up. Findings are discussed with regard to conceptual issues, methodological recommendations, and implications for interventions. PMID- 15887880 TI - Initiation and discontinuation of hormone therapy for menopausal symptoms: results from a community sample. AB - Factors related to initiation and discontinuation of hormonal therapy (HT) for treatment of menopausal symptoms were examined in a community sample of 533 peri/postmenopausal women aged 45-54 by telephone three times. Over 40 variables including: sociodemographic characteristics, clinical reasons to start HT use, HT contraindications, HT attitudes/knowledge, and health behaviors were examined in logistic regression models comparing women who discontinued HT to women who continued using HT over 9 months (discontinuers) and women who initiated HT versus women who never used HT over 9 months (initiators). Increased understanding of HT, confidence, mental health symptoms, perception that menopause is natural, and having gynecological surgery were related to decreased likelihood of HT discontinuation. Increased understanding about risks of HT, vasomotor symptoms, mood symptoms, and having gynecological surgery were related to increased likelihood of HT Initiation. These findings highlight the importance of physicians discussing HT with their patients, particularly because of recent clinical trial developments. PMID- 15887881 TI - Methylmercury, fish consumption, and the precautionary principle. AB - This paper considers several broad issues in the context of probabilistic assessment of the benefits of curtailing mercury (Hg) emissions from U.S. coal fired power plants, based on information developed from recent literature and epidemiology studies of health effects of methylmercury. Exposure of the U.S. population is considered on the national scale, in large part because of recent questions arising from survey and experimental data about the relative importance of local deposition of airborne Hg. Although epidemiological studies have provided useful information, safe levels of Hg exposure remain uncertain, in part because of other dietary considerations in the populations that were studied. For example, much of the seafood consumed in one of the major studies was also contaminated with polychlorinated biphenyls, as are fish taken from some U.S. fresh waters. The primary epidemiological approach involves cross-study comparisons in relation to mean exposures, rather than detailed critiques of individual effects reported in each study. U.S. exposures are seen to be well below the levels at which adverse health effects are reported. This analysis supports the conclusion that unilateral reduction of Hg emissions from U.S. coal fired power plants alone is unlikely to realize significant public health benefits. PMID- 15887882 TI - Source contributions to the mutagenicity of urban particulate air pollution. AB - Using organic compounds as tracers, a chemical mass balance model was employed to investigate the relationship between the mutagenicity of the urban organic aerosol sources and the mutagenicity of the atmospheric samples. The fine particle organic mass concentration present in the 1993 annual average Los Angeles-area composite sample was apportioned among eight emission source types. The largest source contributions to fine particulate organic compound mass concentration were identified as smoke from meat cooking, diesel-powered vehicle exhaust, wood smoke, and paved road dust. However, the largest source contributions to the mutagenicity of the atmospheric sample were natural gas combustion and diesel-powered vehicles. In both the human cell and bacterial assay systems, the combined mutagenicity of the composite of primary source effluents predicted to be present in the atmosphere was statistically indistinguishable from the mutagenicity of the actual atmospheric sample composite. Known primary emissions sources appear to be capable of emitting mutagenic organic matter to the urban atmosphere in amounts sufficient to account for the observed mutagenicity of the ambient samples. The error bounds on this analysis, however, are wide enough to admit to the possible importance of additional mutagenic organics that are formed by atmospheric reaction (e.g., 2 nitrofluoranthene has been identified as an important human cell mutagen in the atmospheric composite studied here, accounting for approximately 1% of the total sample mutagenic potency). PMID- 15887883 TI - Molecular study of thermal immobilization of chromium(VI) with clay. AB - Clay that contains kaolinite has been used extensively as a raw material for manufacturing of bricks and china at 900-1100 degrees C. This study used clay to stabilize the contaminant chromium(VI) [Cr(VI)] through a heating process at 500 1100 degrees C. X-ray absorption spectroscopic results indicated that the 500-900 degrees C heating process transformed hazardous Cr(VI) to nontoxic Cr(III); Cr2O3 was the species detected as most abundant. The 1100 degrees C heating process caused the formation of Cr2SiO5, which was not detected in the samples heated at 500-900 degrees C. Fourier transformed extended X-ray absorption fine structure spectra were fitted by use of WinXAS software. Phase shifts and backscatter(ing) amplitudes for specific atom pairs, based on the crystallographic data for CrO3 and Cr2O3, were theoretically calculated with the FEFF software. The processed XAS data show that the first shell coordination numbers were similar to each other as the temperature was increased from 500 to 900 degrees C and 1100 degrees C, implying that their Cr(III) crystallite size was relatively similar. The interatomic distance between the target center element and the first shell for the 500-1100 degrees C samples was 1.98 A. The Debye-Waller factor for the 1100 degrees C sample was increased compared with the 500 and 900 degrees C samples and probably indicates the formation of Cr2SiO5. PMID- 15887884 TI - Sulfur toxicity and media capacity for H2S removal in biofilters packed with a natural or a commercial granular medium. AB - Two types of media, a natural medium (wood chips) and a commercially engineered medium, were evaluated for sulfur inhibition and capacity for removal of hydrogen sulfide (H2S). Sulfate was added artificially (40, 65, and 100 mg of S/g of medium) to test its effect on removal efficiency and the media. A humidified gas stream of 50 ppm by volume H2S was passed through the media-packed columns, and effluent readings for H2S at the outlet were measured continuously. The overall H2S baseline removal efficiencies of the column packed with natural medium remained >95% over a 2-day period even with the accumulated sulfur species. Added sulfate at a concentration high enough to saturate the biofilter moisture phase did not appear to affect the H2S removal process efficiency. The results of additional experiments with a commercial granular medium also demonstrated that the accumulation of amounts of sulfate sufficient enough to saturate the moisture phase of the medium did not have a significant effect on H2S removal. When the pH of the biofilter medium was lowered to 4, H2S removal efficiency did drop to 36%. This work suggests that sulfate mass transfer through the moisture phase to the biofilm phase does not appear to inhibit H2S removal rates in biofilters. Thus, performance degradation for odor-removing biofilters or H2S breakthrough in field applications is probably caused by other consequences of high H2S loading, such as sulfur precipitation. PMID- 15887885 TI - Enhancing bioremediation of diesel oil and gasoline in soil amended with an agroindustry sludge. AB - This paper presents a study of the bioremediation of diesel oil and gasoline by a series of controlled laboratory tests. Sludge from an agroindustry was used to enhance bioremediation of both gasoline and diesel oil mixed with a soil mass to compare its efficiency with that of a mineral fertilizer. Effects of soil microbiology and soil mixtures were investigated by means of evolution of CO2, microorganism populations at 90 days, pH at 65 and 95 days, mineral nitrogen, and gas chromatographic analysis of the benzene, toluene, methyl tertiary butyl ether, C8, and C9+total aromatics at the end of the experiments. Treatments containing sludge showed better soil conditions after 170 days of treatment (inorganic nitrogen and microbiota activity) compared with gasoline and diesel oil without amendments. Samples had no detectable traces of the measured hydrocarbons at 170 days of treatment. PMID- 15887886 TI - SynGas production from organic waste using non-thermal-pulsed discharge. AB - The purpose of this study was to develop a technology that can convert biogas to synthesis gas (SynGas), a low-emission substituted energy, using a non-thermal pulsed plasma method. To investigate the characteristics of SynGas production from simulated biogas, the reforming characteristics in relation to variations in pulse frequency, biogas component ratio (C3H8/CO2), vapor flow ratio (H2O/total flow rate [TFR]), biogas velocity, and pulse power were studied. A maximum conversion rate of 49.1% was achieved for the biogas when the above parameters were 500 Hz, 1.5, 0.52, 0.32 m/sec, and 657 W, respectively. Under the above conditions, the dry basis mole fractions of the SynGas were as follows: H2 = 0.645, CH4 = 0.081, C2H2 = 0.067, C3H6 = 0.049, CO = 0.008 and C2H4 = 0.004. The ratio of hydrogen to the other intermediates in the SynGas (H2/ITMs) was 3.1. PMID- 15887887 TI - Simulation of the evolution of particle size distributions in a vehicle exhaust plume with unconfined dilution by ambient air. AB - Over the past several years, numerous studies have linked ambient concentrations of particulate matter (PM) to adverse health effects, and more recent studies have identified PM size and surface area as important factors in determining the health effects of PM. This study contributes to a better understanding of the evolution of particle size distributions in exhaust plumes with unconfined dilution by ambient air. It combines computational fluid dynamics (CFD) with an aerosol dynamics model to examine the effects of different streamlines in an exhaust plume, ambient particle size distributions, and vehicle and wind speed on the particle size distribution in an exhaust plume. CFD was used to calculate the flow field and gas mixing for unconfined dilution of a vehicle exhaust plume, and the calculated dilution ratios were then used as input to the aerosol dynamics simulation. The results of the study show that vehicle speed affected the particle size distribution of an exhaust plume because increasing vehicle speed caused more rapid dilution and inhibited coagulation. Ambient particle size distributions had an effect on the smaller sized particles (approximately 10 nm range under some conditions) and larger sized particles (>2 microm) of the particle size distribution. The ambient air particle size distribution affects the larger sizes of the exhaust plume because vehicle exhaust typically contains few particles larger than 2 microm. Finally, the location of a streamline in the exhaust plume had little effect on the particle size distribution; the particle size distribution along any streamline at a distance x differed by less than 5% from the particle size distributions along any other streamline at distance x. PMID- 15887888 TI - Characterization of model error in a simulation of fine particulate matter exposure distributions of the working age population in Helsinki, Finland. AB - Exposure models are needed for comparison of scenarios resulting from alternative policy options. The reliability of models used for such purposes should be quantified by comparing model outputs in a real situation with the corresponding observed exposures. Measurement errors affect the observations, but if the distribution of these errors for single observations is known, the bias caused for the population statistics can be corrected. The current paper does this and calculates model errors for a probabilistic simulation of 48-hr fine particulate matter (PM2.5) exposures. Direct and nested microenvironment-based models are compared. The direct model requires knowledge on the distribution of the indoor concentrations, whereas the nested model calculates indoor concentrations from ambient levels, using infiltration factors and indoor sources. The model error in the mean exposure level was <0.5 microg m(-3) for both models. Relative errors in the estimated population mean were +1% and -5% for the direct and nested models, respectively. Relative errors in the estimated SD were -9% and -23%, respectively. The magnitude of these errors and the errors calculated for population percentiles indicate that the model errors would not drive general conclusions derived from these models, supporting the use of the models as a tool for evaluation of potential exposure reductions in alternative policy scenarios. PMID- 15887889 TI - Optimal fleetwide emissions reductions for passenger ferries: an application of a mixed-integer nonlinear programming model for the New York-New Jersey Harbor. AB - Emissions from passenger ferries operating in urban harbors may contribute significantly to emissions inventories and commuter exposure to air pollution. In particular, ferries are problematic because of high emissions of oxides of nitrogen (NOx) and particulate matter (PM) from primarily unregulated diesel engines. This paper explores technical solutions to reduce pollution from passenger ferries operating in the New York-New Jersey Harbor. The paper discusses and demonstrates a mixed-integer, non-linear programming model used to identify optimal control strategies for meeting NOx and PM reduction targets for 45 privately owned commuter ferries in the harbor. Results from the model can be used by policy-makers to craft programs aimed at achieving least-cost reduction targets. PMID- 15887890 TI - The Steubenville comprehensive air monitoring program (SCAMP): overview and statistical considerations. AB - Average concentrations of particulate matter with an aerodynamic diameter less than or equal to 2.5 microm (PM2.5) in Steubenville, OH, have decreased by more than 10 microg/m3 since the landmark Harvard Six Cities Study associated the city's elevated PM2.5 concentrations with adverse health effects in the 1980s. Given the promulgation of a new National Ambient Air Quality Standard (NAAQS) for PM2.5 in 1997, a current assessment of PM2.5 in the Steubenville region is warranted. The Steubenville Comprehensive Air Monitoring Program (SCAMP) was conducted from 2000 through 2002 to provide such an assessment. The program included both an outdoor ambient air monitoring component and an indoor and personal air sampling component. This paper, which is the first in a series of four that will present results from the outdoor portion of SCAMP, provides an overview of the outdoor ambient air monitoring program and addresses statistical issues, most notably autocorrelation, that have been overlooked by many PM2.5 data analyses. The average PM2.5 concentration measured in Steubenville during SCAMP (18.4 microg/m3) was 3.4 microg/m3 above the annual PM2.5 NAAQS. On average, sulfate and organic material accounted for approximately 31% and 25%, respectively, of the total PM2.5 mass. Local sources contributed an estimated 4.6 microg/m3 to Steubenville's mean PM2.5 concentration. PM2.5 and each of its major ionic components were significantly correlated in space across all pairs of monitoring sites in the region, suggesting the influence of meteorology and long range transport on regional PM2.5 concentrations. Statistically significant autocorrelation was observed among time series of PM2.5 and component data collected at daily and 1-in-4-day frequencies during SCAMP. Results of spatial analyses that accounted for autocorrelation were generally consistent with findings from previous studies that did not consider autocorrelation; however, these analyses also indicated that failure to account for autocorrelation can lead to incorrect conclusions about statistical significance. PMID- 15887891 TI - The Steubenville comprehensive air monitoring program (SCAMP): associations among fine particulate matter, co-pollutants, and meteorological conditions. AB - We determined 24-hr average ambient concentrations of PM2.5 and its ionic and carbonaceous components in Steubenville, OH, between May 2000 and May 2002. We also determined daily average gaseous co-pollutant concentrations, meteorological conditions, and pollen and mold spore counts. Data were analyzed graphically and by linear regression and time series models. Multiple-day episodes of elevated fine particulate matter (PM2.5) concentrations often occurred during periods of locally high temperature (especially during summer), high pressure, or low wind speed (especially during winter) and generally ended with the passage of a frontal system. After removing autocorrelation, we observed statistically significant positive associations between concentrations of PM2.5 and concentrations of CO, NOx, and SO2. Associations with NOx and CO exhibited significant seasonal dependencies, with the strongest correlations during fall and winter. NOx, CO, SO2, O3, temperature, relative humidity, and wind speed were all significant predictors of PM2.5 concentration in a time-series model with external regressors, which successfully accounted for 79% of the variance in log transformed daily PM2.5 concentrations. Coefficient estimates for NOx and temperature varied significantly by season. The results provide insight that may be useful in the development of future PM2.5 reduction strategies for Steubenville. Additionally, they demonstrate the need for PM epidemiology studies in Steubenville (and elsewhere) to carefully consider the potential confounding effects of gaseous co-pollutants, such as CO and NOx, and their seasonally dependent associations with PM2.5. PMID- 15887892 TI - Evaluation of compliance with national legislation on emissions in Portugal. AB - More than 13 years after publication of the first air quality laws in Portugal and more than 10 years after the publication of the respective emission limits, it seems appropriate to analyze the degree of compliance by the Portuguese manufacturing industry. Using the data from emission measurements made regularly by the Instituto de Soldadura e Qualidade, the only officially accredited laboratory according to standard ISO 17025, I analyzed a set of approximately 400 sources in terms of compliance with the emission limits regarding total suspended particulates, sulfur dioxide, nitrogen oxides, and volatile organic compounds. I evaluated compliance through a nondimensional parameter and plotted it versus the emission flow rate to derive conclusions: the results indicate that emission limits are generally met regarding sulfur dioxide and nitrogen oxides but not for the other pollutants considered in this study. However, noncompliance occurs mainly for very low emission flow rates, which suggests some alterations in the emission limits, which are being revised at the moment. These alterations will include the exemption of measurements in minor sources. PMID- 15887893 TI - Mass-size distributions of particulate sulfate, nitrate, and ammonium in a particulate matter nonattainment region in southern Taiwan. AB - Concentrations and distributions of three major water-soluble ion species (sulfate, nitrate, and ammonium) contained in ambient particles were measured at three sampling sites in the Kao-ping ambient air quality basin, Taiwan. Ambient particulate matter (PM) samples were collected in a Micro-orifice Uniform Deposit Impactor from February to July 2003 and were analyzed for water-soluble ion species with an ion chromatograph. The PM1/ PM2.5 and PM1/PM10 concentration ratios at the emission source site were 0.73 and 0.53 and were higher than those (0.68 and 0.48) at the background site because there are more combustion sources (i.e., industrial boilers and traffic) around the emission source site. Mass-size distributions of PM NO3- were found in both the fine and coarse modes. SO4(2-)and NH4+ were found in the fine particle mode (PM2.5), with significant fractions of submicron particles (PM1). The source site had higher PM1/PM10(79, 42, and 90%) and PM1/PM2.5 concentration ratios (90, 58, and 93%) for the three major inorganic secondary aerosol components (SO4(2-), NO3-, and NH4+) than the receptor site (65, 27, and 65% for PM1/PM10, 69, 51, and 70% for PM1/PM2.5. Results obtained in this study indicate that the PM1 (submicron aerosol particles) fraction plays an important role in the ambient atmosphere at both emission source and receptor sites. Further studies regarding the origin and formation of ambient secondary aerosols are planned. PMID- 15887894 TI - Preliminary economic analysis of poultry litter gasification option with a simple transportation model. AB - Several environmental issues are related to the disposal of poultry litter. In an effort to provide a more environmentally friendly alternative than landfill disposal or spreading as a soil amendment, work has been carried out previously at the University of Tennessee Space Institute (UTSI). This past UTSI work was concentrated on developing a catalytic steam gasification concept to produce energy from poultry litter. In the past UTSI studies, preliminary design and economics for a stationary, centralized gasification plant capable of processing approximately 100 ton/day of poultry litter were developed. However, in this preliminary design the economic impact of transporting litter to a centralized gasification plant location was not addressed. To determine the preliminary impact of transporting the poultry litter on the overall economics of this energy conversion plant design, a simple transportation model was developed. This model was used in conjunction with the earlier plant design prepared at UTSI to determine the economic feasibility of a centralized, stationary poultry litter gasification plant. To do so, major variables such as traveling distance, plant feed rate (or capacity), fluctuations in the sales price of the product gas (that means value of the energy), population density of poultry farms, impact of tipping fees, and cost of litter were varied. The study showed that for plant with a capacity of 1000 ton/day to be able to withstand several changes in economic conditions and sustain itself, the poultry farm density would need to be approximately 0.3 houses/mi2. Smaller plants would need either a higher energy price or some kind of subsidy to be economically feasible. PMID- 15887895 TI - Analysis of air quality observations with the aid of the source-receptor relationship approach. AB - In this study, an attempt was made to analyze time series of air quality measurements (O3, SO2, SO4(2-), NOx) conducted at a remote place in the eastern Mediterranean (Finokalia at Crete Island in 1999) to obtain concrete information on potential contributions from emission sources. For the definition of a source receptor relationship, advanced meteorological and dispersion models appropriate to identify "areas of influence" have been used. The model tools used are the Regional Atmospheric Modeling System and the Lagrangian-type particle dispersion model (forward and backward in time), with capabilities to derive influence functions and definition of "areas of influence." When high levels of pollutants have been measured at the remote location of Finokalia, particles are released from this location (receptor) and traced backward in time. The influence function derived from particle distributions characterizes dispersion conditions in the atmosphere and also provides information on potential contributions from emission sources within the modeling domain to this high concentration. As was shown in the simulation results, the experimental site of Finokalia in Crete is influenced during the selected case studies, primarily by pollutants emitted from the urban conglomerate of Athens. Secondarily, it is influenced by polluted air masses arriving from Italy and/or the Black Sea Region. For some specific cases, air pollutants monitored at Finokalia were possibly related to war activities in the West Balkan Region (Kosovo). PMID- 15887896 TI - Methods for injury surveillance in international cricket. AB - BACKGROUND: The varying methods of cricket injury surveillance projects have made direct comparison of published studies in this field impossible. METHODS: A consensus regarding definitions and methods to calculate injury rates in cricket was sought between researchers in this field. This was arrived at through a variety of face-to-face meetings, email communication and draft reviews between researchers from six of the major cricket-playing nations. RESULTS: It is recommended that a cricket injury is defined as any injury or other medical condition that either: (a) prevents a player from being fully available for selection for a major match or (b) during a major match, causes a player to be unable to bat, bowl or keep wicket when required by either the rules or the team's captain. Recommended definitions for injury incidence (for matches, training sessions and seasons) and injury prevalence are also provided. It is proposed that match injury incidence is calculated using a denominator based on a standard time estimated for player exposure in matches, for the purposes of simplicity. This will allow all injury surveillance systems, including those with limited resources, to make calculations according to a standard definition. CONCLUSION: The consensus statement presented provides a standard which, if followed, allows meaningful comparison of injury surveillance data from different countries and time periods, which will assist in the possible identification of risk factors for injury in cricket. PMID- 15887897 TI - Physiological and performance benefits of halftime cooling. AB - This study examined the effect of a 10-min, halftime cooling application on physiological and psychological parameters known to affect performance. Fourteen volunteers (10 male, 4 female) completed two randomised trials 48 hr to 7 days apart. Trials consisted of a 1-hr cycling protocol: 30 min at 75% VO2max followed by 10 min cooling (application of a cooling jacket) or passive recovery (control), and a second 30-min exercise bout consisting of 20 min at 75% VO2max, immediately followed by a 10-min maximal effort, where work was measured as energy expended (kJ). Performance of the 10-min maximal intensity phase tended to improve (171.5 +/- 30.4 kJ vs 165.4 +/- 29.2 kJ, p = 0.087) following the cooling trial. Heart rate during the 5th min of the maximal effort, (183 +/- 9 beats.min( 1) vs 180 +/- 7 beats.min(-1), p = 0.024), blood lactate concentration at 6 min post-exercise (9.3 +/- 3.1 mmolxL(-1) vs 7.9 +/- 3.2 mmolxL(-1), p = 0.007), rating of perceived exertion at the 20th min post-halftime recovery (15 +/- 2 vs 16 +/- 2, p = 0.042), and subjective rating of feelings and emotions differed between the cooling and control conditions. Sweat loss, core and mean skin temperature and rating of thermal sensation failed to differ significantly between conditions. Halftime cooling tended to result in greater aerobic performance. Psychological assessment revealed a dramatic placebo effect from the cooling application confounding these results. Furthermore, the cooling intervention failed to induce any significant thermoregulatory effects. PMID- 15887898 TI - Are athletes able to self-select their optimal warm up? AB - This study aimed to examine whether athletes are able to self-select their optimal warm up and to propose a methodological approach in investigating the effects of warm up on performance. Nine male subjects underwent a free field warm up (FWU) at a self-selected intensity and duration during which heart rate (HR) and rectal temperature (Tre) were monitored. The intensity of this warm up was subsequently estimated from the HR obtained during an incremental test to determine maximal power (Pmax). Performance (cycle time to exhaustion at Pmax), HR and Tre, were then examined following either: NWU (no warm up); RWU (reference warm up based on FWU); RWU-10 (warm up intensity diminished by 10% compared to RWU); and RWU+10 (warm up intensity increased by 10% compared to RWU). Results showed no significant difference in HR (P = 0.37) and Tre increase (P= 0.77) between FWU and RWU. Performance improvement after warm up conditions gave RWU (56%; ie, 5/9 subjects) >RWU-10 (33%; ie, 3/9) >RWU+10 (11%; ie, 1/9) >NWU with significant differences between RWU and NWU (P < 0.01); RWU and RWU+10 (P < 0.01); RWU-10 and NWU (P < 0.01). A warm up intensity ranging from 54-72% Pmax, and inducing an increase in heart rate to 80 +/- 6% HRmax, was found to be optimal. While most athletes were able to self-determine the intensity of their optimal warm up, for others there is still a need for control. PMID- 15887899 TI - The physiology of falling: assessment and prevention strategies for older people. AB - Balance calls upon contributions from vision, peripheral sensation, vestibular sense, muscle strength, neuromuscular control and reaction time. With increased age, there is a progressive loss of functioning of these systems and an increased likelihood of falls. Falls can mark the beginning of a decline in function and independence and are the leading cause of injury-related hospitalisation in older people. By using simple tests of vision, leg sensation, muscle strength, reaction time and standing balance, it is possible to identify accurately older people at risk of falls and assess intervention outcomes. This approach overcomes the limitations associated with traditional methods of assessing falls risk via medical diagnoses, including varied severity between individuals. Using a physiological approach provides information at the impairment and functional capacity levels to assist in understanding falls and developing and evaluating optimal falls prevention strategies for older people. PMID- 15887900 TI - Upper body aerobic fitness comparison between two groups of competitive surfboard riders. AB - The aim of the present study was to evaluate and compare the upper-body aerobic fitness characteristics in 2 groups of competitive surfers with different performance levels. Thirteen male competitive surfers performed an incremental dry-land board paddling test to determine specific peak oxygen uptake (VO2peak), peak power output (Wpeak) and the exercise intensity (%VO2peak) that elicits a blood lactate concentration of 4 mmol x L(-1) (LT4). As a measure of surfing performance, surfers were ranked according to their competitive season performance (RANK) and divided into 2 groups based on their performance level; European top-level competitive surfers (ELS) (n = 7) and regional level competitive surfers (RLS) (n = 6). ELS reached significantly higher values than RLS for Wpeak (154.71 +/- 36.82 W vs. 117.70 +/- 27.14 W: P = 0.04) and LT4 (95.18 +/- 3.42 %VO2peak vs. 88.89 +/- 5.01 %VO2peak; P = 0.02) but not VO2peak (3.34 +/- 0.31 L x min(-1) vs. 3.40 +/- 0.37 L x min(-1); P = 0.77). Spearman rank order correlation analysis revealed that RANK was inversely correlated with Wpeak (r = -0.65, P = 0.01) and LT4 (r = -0.58, P = 0.03). These findings identify that better surfers have higher upper body aerobic fitness scores. PMID- 15887901 TI - The development of a test of reactive agility for netball: a new methodology. AB - The purpose of this study was to present a new methodology for the measurement of agility for netball that is considered more ecologically valid than previous agility tests. Specifically, the agility performance of highly-skilled (n = 12), moderately-skilled (n = 12) and lesser-skilled players (n = 8) when responding to a life-size, interactive video display of a netball player initiating a pass was compared to a traditional, pre-planned agility movement where no external stimulus was present. The total movement times and decision times of the players were the primary dependent measures of interest. A second purpose of the research was to determine the test-retest reliability of the testing approach. Results revealed significant differences existed between the 2 test conditions demonstrating that they were measuring different types of agility. The highly skilled group was significantly faster in both the reactive and planned test conditions relative to the lesser-skilled group, while the moderately-skilled group was significantly faster than the lesser-skilled group in the reactive test condition. The decision time component within the reactive test condition revealed that the highly-skilled players made significantly faster decisions than the lesser-skilled players. It is reasoned that it is this decision-making component of reactive agility that contributes to the significant differences between the two test conditions. The testing approach was shown to have good test retest reliability with an intra-class correlation of r = .83. PMID- 15887902 TI - Reliability and validity of physical fitness field tests for adults aged 55 to 70 years. AB - The aim of this study was to examine the reliability and validity of field tests for assessing physical function in mid-aged and young-old people (55-70 y). Tests were selected that required minimal space and equipment and could be implemented in multiple field settings such as a general practitioner's office. Nineteen participants completed 2 field and 1 laboratory testing sessions. Intra-class correlations showed good reliability for the tests of upper body strength (lift and reach, R= .66), lower body strength (sit to stand, R = .80) and functional capacity (Canadian Step Test, R= .92), but not for leg power (single timed chair rise. R = .28). There was also good reliability for the balance test during 3 stances: parallel (94.7% agreement), semi-tandem (73.7%), and tandem (52.6%). Comparison of field test results with objective laboratory measures found good validity for the sit to stand (cf 1RM leg press, Pearson r= .68, p < .05), and for the step test (cf PWC140, r = -.60, p < .001), but not for the lift and reach (cf 1RM bench press, r = .43, p > .05), balance (r = -.13, -.18, .23) and rate of force development tests (r = -.28). It was concluded that the lower body strength and cardiovascular function tests were appropriate for use in field settings with mid-aged and young-old adults. PMID- 15887903 TI - A decrease in the association between the physical activity patterns of Australian parents and their children; 1985-1997. AB - The present study investigated the interactions between parents' and children's physical activity levels by examining whether or not parents who exercise have children who participate in sport. Of primary interest was an investigation of trends in these interactions over time. Information was collected from 10-13 y old children in 1985 (n = 2463) and then again in 1997-99 (n = 1469), about their sports participation and their perceptions of parents' exercise habits. Boys' participation in at least one sport declined from 87% in 1985 to 76% in 1997/1999 while, among girls, participation fell from 80% to 71%. According to their children's perceptions, mothers exercising regularly fell from 36% to 31% between surveys, while fathers exercising regularly fell from 39% to 32%. Interactions between parents' and children's exercise and sports behaviours were examined employing chi-square analysis techniques. Results showed gender-specific relationships for the 1985 sample, such that active fathers were associated with increased participation in sports by boys, and inactive mothers were associated with less participation in sports by girls. These interactions seemed to diminish over time. It is possible that changes in social structures during this time may be affecting familial behaviour relationships, such as the role modelling of active behaviours. PMID- 15887904 TI - Hip joint range of motion reduction in sports-related chronic groin injury diagnosed as pubic bone stress injury. AB - Traumatic osteitis pubis is a non-specific entity that relates to chronic groin injury and has recently been described as being akin to a pubic bone stress injury. It is uncertain whether or not reduction of hip joint range of motion occurs in traumatic osteitis pubis. The purpose of this study was to establish whether there is a reduction of hip range of motion in athletes who have chronic groin injury diagnosed as pubic bone stress injury. A case-control study was performed whereby 89 Australian Rules footballers underwent, with clinical history unknown, clinical and MRI examination of the groin region. Clinical criteria (pain with tenderness) and MR-criteria (pubic bone marrow oedema) were used for diagnosis of pubic bone stress injury. End-range internal and external rotation hip motion was measured using a goniometer. Athletes with and without symptoms were compared, as were athletes with current symptoms with athletes who had recovered from their groin pain episode. Chronic groin injury was diagnosed in 47 athletes with 37 having pubic bone stress injury. Thirteen athletes had previous groin injury. A reduction of internal and external hip range of motion was demonstrated in athletes with pubic bone stress injury (p < 0.05) and in athletes who had current symptoms compared to those who had recovered from their groin pain episode (p < 0.05). A reduction in hip range of motion was evident in athletes with chronic groin injury diagnosed as pubic bone stress injury. There may be a role for increasing hip range of motion in rehabilitation. PMID- 15887905 TI - Does a balance deficit persist in Australian Football players with previous lower limb ligament injury? AB - A history of lower limb ligament injury is a commonly-cited risk factor for another similar injury. During the acute phase of injury, there is a balancing skill deficit in the injured limb. It has been unclear as to whether this deficit persists in the medium-to-long term for previously injured Australian footballers, contributing to the risk of re-injury. This study compared the balance ability of footballers with and without previous lower limb ligament injury and, for previously injured players, the balance ability of the previously injured limb to the opposite uninjured limb. A total of 216 players from 6 teams from the Australian Football League were tested. The balance task comprised stepping on to a foam mat on top of a force plate and maintaining one-legged balance. The subjects were divided into 4 groups based on their injury history: all ankle injuries to only one limb, recent ankle injuries to only one limb (within the last 12 months), knee ligament injury only to one limb, and no previous ankle or knee ligament injury. Statistical analysis revealed that there was no significant difference between the balance scores of any of the previously injured players and those with no previous lower limb ligament injury. There was no significant difference between the balance score of the previously injured limb with the opposite uninjured limb. It appears that a balance deficit does not persist in Australian Football players with previous lower limb ligament injury. PMID- 15887906 TI - Unprotected eyes in squash: not seeing the risk of injury. AB - The use of appropriate eyewear in squash can protect the eyes against injury. However, few adult squash players adequately protect their eyes against potential severe injuries. We describe the characteristics of non-users of protective eyewear and examine predictors of appropriate eyewear use. Self-report surveys of adult players were conducted in metropolitan Melbourne, Australia. Information on players' knowledge, behaviours and attitudes associated with protective eyewear use was collected, in addition to player demographic data. Appropriate eyewear was defined as Standards-approved polycarbonate lens eyewear. The majority 92.2% of players did not adequately protect their eyes while playing squash. Significant predictors of reported eyewear use were: previous eye injury: playing squash on average more than 2 hr per wk: having played for more than 20 y; and having more favourable attitudes towards eye safety in squash. The significant predictors of appropriate eyewear use were: being female; previous eye injury; playing squash on average more than 2 hr per week; and having more favourable attitudes towards eye safety in squash. Understanding the characteristics of both users and non-users of appropriate eye protection in squash is essential for informing future prevention strategies. PMID- 15887907 TI - The fair sex? Foul play among female rugby players. AB - Rugby, a full contact sport, exposes participants to a high risk of injury. While several studies have explored injuries among male rugby players, few have investigated injuries among females. We conducted a cross-sectional study of United States of America (USA) female rugby players to assess the players' perception of foul play and the referee response to foul play and to evaluate the association between players' perception of foul play and injury. An anonymous, self-administered questionnaire reporting injury status, history of player perceived foul play and referee response was administered to 258 players recruited at a women's rugby tournament. The overall rate of injury was 4.4 injuries/100 matches, 0.2 injuries/100 practices and 1.4 injuries/100 total rugby exposures (matches and practices), with 107 (41.5%) players classified as injured. While 16.5% of players admitted to perpetrating foul play without an assessed penalty and 13.8% to being penalised for foul play, a smaller proportion reported being sent to the 'sin bin' (temporarily removed from play) or being ejected from a match (3.3% and 1.3% respectively). Of the 107 injured, 24.3% believed they had been injured as a result of foul play. Among all 258 players, self-perception of having been hurt due to unpenalised foul play was associated with study-defined injury (OR = 2.4, 95% CI = 1.0-5.9, p = 0.046). To make the sport safer, efforts should be made to minimise foul play. Suggested preventive methods include educating referees, coaches and players about the prevalence of foul play in women's rugby and the association between foul play and injury. PMID- 15887908 TI - Influence of the limited interchange rule on injury rates in sub-elite Rugby League players. AB - This study investigated the incidence of injury in sub-elite Rugby League players before and after the introduction of the limited interchange rule. The incidence of injury was prospectively studied in one sub-elite Rugby League club over three competitive seasons. Two seasons were played under the unlimited interchange rule, while the third season was played under the limited interchange rule. Players participated in 91 matches under the unlimited interchange rule and 44 matches under the limited interchange rule. Injury was defined as any pain, disability or injury that occurred as a result of a competition game that caused the player to miss a subsequent game. The relative risk (RR) of injury significantly decreased (RR = 0.70 [0.65 to 0.75], P<0.05) following the introduction of the limited interchange rule, with the incidence of injury decreasing from 72.5 (58.2 to 86.8) per 1000 playing hr under the unlimited interchange rule to 51.0 (33.8 to 68.1) per 1000 playing hr under the limited interchange rule. The risk of sustaining thigh and calf injuries (RR= 0.27 [0.12 to 0.59], P < 0.05), muscular strains (RR= 0.23 10.17 to 0.31]., P<0.05), and high intensity running injuries (RR= 0.16 [0.04 to 0.691, P<0.05) was significantly reduced following the introduction of the limited interchange rule. These findings demonstrate that the risk of injury in sub-elite Rugby League players is significantly reduced following the introduction of the limited interchange rule. The reduced injury risk may reflect a fatigue-induced reduction in match speed and impact forces associated with physical collisions and tackles. PMID- 15887909 TI - [Systemic thrombolysis after cerebral infarct: every minute counts]. PMID- 15887910 TI - [Early mobilization of patients with increased intracranial pressure: kinesthetics for the benefit of patients and nurses]. PMID- 15887911 TI - [Influence of behavior on motoricity after a stroke: learning mobility anew]. PMID- 15887912 TI - [Logopedics in neurologic rehabilitation: properly supporting patients in "home work"]. PMID- 15887913 TI - [Ergotherapy--a "routine" therapy? Practice does not end in the treatment room]. PMID- 15887914 TI - [Patients with pusher symptoms: finding the objective perpendicular line]. PMID- 15887915 TI - [Nursing patients in coma vigilans in community residences: stimulating all senses]. PMID- 15887916 TI - [Neuro Development Treatment under discussion: "Patients should concentrate more on everyday skills" (interview by Katrin Balzer)]. PMID- 15887917 TI - [Neuro Development Treatment under discussion: not throwing the child out with the bath water]. PMID- 15887918 TI - [Stroke units: critical reflection on a success concept]. PMID- 15887919 TI - [Respite for family caregivers: "I am already planning to do this a while longer"]. PMID- 15887920 TI - [Implementing the nursing care legislation in Lower Saxony: first steps on a long path]. PMID- 15887921 TI - [Clinical treatment pathways--2: The path is the goal--and an optimized organizational methodology]. PMID- 15887922 TI - [Promoting mental health at the work site: support program can improve industry climate]. PMID- 15887923 TI - [Coaching for residential facility management--1: An innovate form of personal development?]. PMID- 15887924 TI - [Increased pressure]. PMID- 15887925 TI - [Testing guideline oriented education in North Rhine Westphalia. Results from Recklinghausen Central Nursing School of the Knappschaft Hospital]. PMID- 15887926 TI - Economics 101 for the nonbelievers. PMID- 15887927 TI - Rheumatoid arthritis: an update. AB - Rheumatoid arthritis is a chronic multi-system disorder of unknown etiology with most common symptoms being persistent inflammatory synovitis, usually involving peripheral joints in a symmetrical pattern. Improved understanding of the molecular mechanisms involved in the pathogenesis of rheumatoid arthritis has led to long awaited, highly effective treatments for this disorder. PMID- 15887928 TI - Advances in biotechnology: radiology. PMID- 15887929 TI - We shall fight them on the beaches. PMID- 15887931 TI - Management of inflammatory skin conditions in Australian general practice. PMID- 15887932 TI - Eczema--practical management issues. AB - BACKGROUND: Eczema is a common, and at times challenging, condition to manage. It often involves an irritable child, concerned parents and numerous return visits. A variety of mainstream and alternative treatments can confuse both the clinician and patient. OBJECTIVE: This article aims to refresh readers by reviewing evidenced based treatment protocols, and exploring some of the new and evolving treatments in eczema. DISCUSSION: Managing eczema is a multifocal task involving a variety of medicinal and practical approaches. Education of parents and carers is critical. Having a clear plan that can be adapted to each patient's needs is helpful for all involved, with management focussing on control rather than cure. Simple emollients and preventive measures are used to reduce heat, dryness and prickling of the skin. Topical corticosteroids remain the gold standard in the treatment of the inflammation of eczema. However, combining topical steroids with the new calcineurin inhibitors, wet dressings and behavioural modification should increase the time interval between exacerbations. PMID- 15887933 TI - Occupational dermatoses. AB - BACKGROUND: Occupational contact dermatitis is a common condition often assumed to be 'part of the job'. OBJECTIVE: This article explores common causes of occupational contact dermatitis and details high risk occupations utilising local data. Tips for improved understanding and management of occupational contact dermatitis are also presented. DISCUSSION: General practitioners have an important part to play in the recognition and management of occupational contact dermatitis. Early diagnosis and treatment can improve the outcome in patients with occupational contact dermatitis. PMID- 15887934 TI - Cutaneous manifestations of systemic disease. AB - BACKGROUND: While most patients who present with a rash have no associated systemic illness, many systemic illnesses have skin manifestations at some stage. OBJECTIVE: This article uses case vignettes to illustrate a problem oriented approach to five commonly presented skin conditions that have common and varied systemic associations. A logical sequence of management for each case is provided. DISCUSSION: Often the skin manifestations of systemic disease are vague and nonspecific such as the toxic erythema that might follow a viral infection or a drug eruption. Sometimes it is the systemic manifestations that are vague and nonspecific, while the skin manifestations are highly specific and define the illness. There is currently no useful classification for cutaneous manifestations of systemic disease. PMID- 15887935 TI - Patient education. Management of infantile eczema. PMID- 15887936 TI - Fatal adverse drug reaction to trimethoprim. AB - BACKGROUND: Trimethoprim is the antibiotic of choice for treating uncomplicated community acquired urinary tract infections. However, before prescribing any drug it is vital to obtain a detailed drug history to exclude possible drug allergy. OBJECTIVE: We present the case of a fatal adverse drug reaction due to trimethoprim. We discuss some of the problems that led to the patient's death, and consider how to prevent similar events. DISCUSSION: Re-exposing a patient to a drug suspected of causing an adverse reaction is associated with considerable risk of morbidity and mortality. A detailed history is vital to correctly diagnose drug allergies but, even so, it is not always possible to identify the causative agent. Following an adverse drug reaction, health professionals must take appropriate steps to avoid inadvertently re-prescribing the culprit drug. This is essential to avoid the tragic outcome highlighted in this case. PMID- 15887937 TI - Procedures in primary care dermatology. AB - BACKGROUND: General practitioners can provide a range of diagnostic and treatment procedures for patients with dermatological problems. OBJECTIVE: This article reviews the use of Wood's light, diascopy, skin scrapings for microscopy, skin biopsy, cauterisation, doppler measurement of ankle-brachial pressure index, and multi-layer compression bandages for venous ulcers. DISCUSSION: Adequate training and proper equipment is essential for commonly neglected areas including informed written consent, good communication with the histopathologist, a system for handling specimens and reports, action plans for laboratory results, and adequate training and protection for auxiliary staff. PMID- 15887938 TI - Management of benign breast conditions. Part 3--Other breast problems. AB - This is the third article in a series of breast disorders with an emphasis on diagnosis and management in the general practice setting. This article discusses conditions that, although less frequently seen in general practice, pose challenges in diagnosis and management. PMID- 15887939 TI - Patient education. Nipple discharge. PMID- 15887940 TI - Assessing fitness to drive--part 1. AB - BACKGROUND: The requirement for general practitioners to write reports about their patients' fitness to drive will increase as the population ages and licensing criteria change. It is important that GPs understand the medical and legal issues involved in this important area of public health. OBJECTIVE: This two part article discusses the rationale behind assessing fitness to drive and briefly summarises several medical conditions that commonly give rise to problems. This information will help GPs understand the decision making process regarding this sensitive issue, and improve the quality of medical reports. Adequate assessment and reporting can help patients avoid becoming involved in traffic crashes, and the doctor from becoming involved in court appearances. DISCUSSION: Specific medical conditions discussed in part one are epilepsy, diabetes, and cardiovascular disease. Guidelines are based on currently available evidence regarding the effects of medical conditions on driving and are subject to regular review as new information becomes available. PMID- 15887941 TI - Meeting the needs of patients with cancer--a GP guide to support services. AB - BACKGROUND: A diagnosis of cancer places huge psychological and physical demands on individuals. Early stage cancer patients will frequently experience depression and elevated levels of anxiety. OBJECTIVE: This article summarises the available cancer services, categorised to address patient needs, and suggests suitable times to provide this information to patients. DISCUSSION: Cancer patients report a moderate to high need for help regarding their ability to cope with their illness and its treatment, emotional support, information provision and relations with staff at treatment centres. General practitioners are involved in cancer care, from initial screening to palliative care. Therefore it is essential that GPs are up-to-date with the available cancer services to assist in meeting patient needs in cancer care. PMID- 15887942 TI - The hazards of horse riding. PMID- 15887943 TI - Standards and performance--attainment and maintenance of professional capabilities. AB - The nature of The Royal Australian College of General Practitioners (RACGP) examination came under scrutiny in a recent debate among RACGP members, some of who suggested exploring an alternative pathway for assessment linked to continuing medical education. This article outlines key issues underpinning the examination that is part of the requirements for attaining Fellowship of the RACGP (FRACGP). It provides an overview of the theory and practice of assessment for general practice. The RACGP examination has an international reputation for quality, validity and reliability, a reason why the RACGP has been asked to assist many others in establishing and/or reviewing their own examination processes. PMID- 15887944 TI - Consent to medical treatment. AB - Obtaining patient consent is good medical practice and a legal necessity. This article examines the duty of general practitioners to obtain consent from patients for medical interventions and outlines the process of obtaining consent. PMID- 15887945 TI - Government involvement in general practice. PMID- 15887946 TI - General practitioner, specialist providers case conferences in palliative care- lessons learned from 56 case conferences. AB - OBJECTIVE: To describe the utility and acceptability to general practitioners and palliative care staff of case conferences in palliative care. METHOD: Research focussed on case conferences conducted between GPs and staff of three specialist palliative care units (in an inner urban, outer metropolitan and regional setting), at the time of referral of patients to the service. Telephone interviews were conducted with all GPs who participated in a case conference, and focus groups were conducted with palliative care staff. RESULTS: For most GPs, case conferences by teleconference were a time effective and immediate means of information transfer. The best instances for a conference were at time of patient referral, time of discharge to the community, or where the case was complex. General practitioners appreciated access to multiple professionals simultaneously. Workload pressures were a drawback of participation for both GPs and specialists. Palliative care team members thought case conferences gave GPs an appreciation of a team approach, and reduced professional isolation. The usefulness of the case conferences depended on the willingness of the GP to participate. General practitioners would participate again provided they did not have to organise the case conference. Specialist staff were concerned by the financial cost of organising case conferences. DISCUSSION: Case conferences provide useful information exchange between GPs and specialist staff, and are acceptable to both parties. Much depends on the individual GPs attitude toward participation, as well as the timing of the conferences in the course of the patient's illness. Organisation needs to be a task of the specialist units, who would need administrative support to organise them. PMID- 15887947 TI - Australia's Cancer Helpline--an audit of utility and caller profile. AB - BACKGROUND: The Cancer Helpline is a telephone information and support service operated throughout Australia. METHODS: Data from all calls received during the months of July, August and September from 2001 to 2003 were collected and analysed. RESULTS: Over 76,000 contacts were made, mostly from women (79%). Almost half of all calls were from people with general inquiries (rather than cancer patients, their families, or health professionals). The most common cancer type discussed was breast cancer, (26% of all calls, 42% of calls from people with cancer). Diagnosed cancer patients most frequently called about treatment/management and practical issues. People with general inquiries called to obtain information about the prevention and early detection of cancer. DISCUSSION: The helpline can complement the role of other information and support providers for people living with or concerned about cancer. PMID- 15887948 TI - Management of impacted ear wax--an observational report. PMID- 15887949 TI - Patients' views on chronic illness and its care in general practice. AB - BACKGROUND: General practitioners have described barriers to the uptake of incentives such as the Enhanced Primary Care items introduced to support care of chronic illness. However patients' attitudes toward chronic illness and planned care are not known. METHODS: A qualitative study of adult patients with chronic illnesses using semi-structured interviews and a focus group, examining their experience of chronic illness and their attitudes toward health care. RESULTS: Sixteen patients were interviewed; five participated in the focus group. Patients believed that their illnesses were permanent and progressive, and that GPs had little influence over their prognosis. They preferred to access GPs for acute problems rather than anticipatory care, and most could not see a need for care planning. DISCUSSION: Patient preferences for care delivery may be in conflict with the current reform agenda focussing on anticipatory care provision. Efforts to reform chronic illness care may falter unless this is addressed. PMID- 15887950 TI - Computer use in general practice consultations. PMID- 15887951 TI - The next generation of phosphodiesterase inhibitors: structural clues to ligand and substrate selectivity of phosphodiesterases. PMID- 15887952 TI - Structure-based virtual screening for plant-based ERbeta-selective ligands as potential preventative therapy against age-related neurodegenerative diseases. AB - ERbeta has been associated with estrogen-induced promotion of memory function and neuronal survival. Based on the optimized complex structure of human ERbeta LBD bound with genistein, computer-aided structure-based virtual screening against a natural source chemical database was conducted to determine the occurrence of plant-based ERbeta-selective ligands. Twelve representative hits derived from database screening were assessed for their binding profiles to both ERs, three of which displayed over 100-fold binding selectivity to ERbeta over ERalpha. PMID- 15887953 TI - Synthesis and biological evaluation of benzodioxanylpiperazine derivatives as potent serotonin 5-HT(1A) antagonists: the discovery of Lecozotan. AB - A series of benzodioxanylpiperazine derivatives possessing a 4-aryl amide substituent was prepared and evaluated for 5-HT(1A) affinity and functional antagonist activity in vitro and in vivo. All of the compounds in this series possessed high affinity for the human 5-HT(1A) receptor and many displayed potent antagonist activity in vitro and varying degrees of intrinsic activity in vivo. Compound 11c (Lecozotan) was selected for further development and is currently in clinical trials. PMID- 15887954 TI - Oxazine 170 induces DNA:RNA:DNA triplex formation. AB - The oxazine dye (Oxazine 170) was found to induce formation of a hybrid triplex structure, poly rA:(poly dT)(2), under solution conditions in which the triplex would not otherwise form. Formation of the hybrid triplex is driven by the structural selective binding of oxazine 170 to poly rA:(poly dT)(2). Oxazine 170 serves as a lead compound for the design of new compounds that can modulate triplex formation. PMID- 15887955 TI - Varenicline: an alpha4beta2 nicotinic receptor partial agonist for smoking cessation. AB - Herein we describe a novel series of compounds from which varenicline (1, 6,7,8,9 tetrahydro-6,10-methano-6H-pyrazino[2,3-h][3]benzazepine) has been identified for smoking cessation. Neuronal nicotinic acetylcholine receptors (nAChRs) mediate the dependence-producing effects of nicotine. We have pursued alpha4beta2 nicotinic receptor partial agonists to inhibit dopaminergic activation produced by smoking while simultaneously providing relief from the craving and withdrawal syndrome that accompanies cessation attempts. Varenicline displays high alpha4beta2 nAChR affinity and the desired in vivo dopaminergic profile. PMID- 15887956 TI - Discovery of the first potent, selective 5-hydroxytryptamine1D receptor antagonist. AB - A series of 5-(piperidinylethyloxy)quinoline 5-hydroxytryptamine(1D) (5-HT(1D)) receptor antagonists have been discovered from elaboration of the series of dual 5-hydroxytryptamine(1)-selective serotonin reuptake inhibitors (5HT(1)-SSRIs) reported previously. This is the first report of highly potent, selective antagonists for the 5-HT(1D) receptor, which represents an extremely useful set of pharmacological tools for further understanding the roles of the 5-HT(1) receptor subtypes. PMID- 15887957 TI - Synthesis and structure-activity-relationships of 1H-imidazo[4,5-c]quinolines that induce interferon production. AB - 1H-Imidazo-[4,5-c]quinolines were prepared while investigating novel nucleoside analogues as potential antiviral agents. While these compounds showed no direct antiviral activity when tested in a number of cell culture systems, some demonstrated potent inhibition of virus lesion development in an intravaginal guinea pig herpes simplex virus-2 assay. We have determined that the in vivo antiviral activity can be attributed to the ability of these molecules to induce the production of cytokines, especially interferon (IFN), in this model. Subsequently, we found that the compounds also induce in vitro production of IFN in human peripheral blood mononuclear cells (hPBMCs). The in vitro results reported herein and the in vivo results reported previously led to the discovery of imiquimod, 26, which was developed as a topical agent and has been approved for the treatment of genital warts, actinic keratosis, and superficial basal cell carcinoma. PMID- 15887958 TI - Effect on K(ATP) channel activation properties and tissue selectivity of the nature of the substituent in the 7- and the 3-position of 4H-1,2,4 benzothiadiazine 1,1-dioxides. AB - The present work explored 3-alkylamino-4H-1,2,4-benzothiadiazine 1,1-dioxides diversely substituted in the 7-position. Those compounds, structurally related to previously described potassium channel openers such as the benzothiadiazine dioxide BPDZ 73, were tested as putative K(ATP) channel activators on the pancreatic endocrine tissue and on the vascular smooth muscle tissue. The nature of the substituent introduced in the 7-position as well as the nature of the alkylamino side chain in the 3-position strongly affected both potency and tissue selectivity of 4H-1,2,4-benzothiadiazine 1,1-dioxides. Thus, compounds bearing in the 7-position a methyl or a methoxy group or devoid of a substituent in this position, and bearing an ethyl, an isopropyl, or a cyclobutylamino group in the 3 position were found to be potent and selective inhibitors of insulin release from rat pancreatic B-cells (i.e. 10a, 10b, 12b, 12d, 22c). In contrast, 3-alkylamino 7-trifluoromethyl- (20a-c) and 3-alkylamino-7-pentyl-4H-1,2,4-benzothiadiazine 1,1-dioxides (11a,b) expressed a marked myorelaxant activity on rat aorta ring. Among the latter compounds, the 3-alkylamino-7-pentyl derivative (11a) showed a clear selectivity for the vascular smooth muscle tissue. The present work gives new insights into the role of the substituent in both the 7- and the 3-position for the design of 4H-1,2,4-benzothiadiazine 1,1-dioxide potassium channel openers exhibiting different tissue selectivity profiles. PMID- 15887959 TI - Application of phosphoramidate pronucleotide technology to abacavir leads to a significant enhancement of antiviral potency. AB - We report the first application of pronucleotide (ProTide) technology to the antiviral agent abacavir (Ziagen), used for the treatment of HIV infection. The phenylmethoxyalaninyl phosphoramidate of abacavir was prepared in good yield in one step. Also prepared was the corresponding phosphoramidate of the guanine nucleoside analogue "carbovir". The antiviral profile of each of the parent nucleosides was compared to that of the phosphoramidate ProTides. A significant (28- to 60-fold) increase in anti-HIV potency was noted for the ProTide of abacavir but not for that of carbovir. These findings were in agreement with the markedly higher (ca. 37-fold) levels of carbovir triphosphate that are formed in CEM cells upon response to the abacavir ProTide compared with the parent abacavir compound. In contrast the anti-HBV potency of both abacavir and carbovir were improved (10- and 20-fold, respectively) by ProTide formation. As in CEM cells, the abacavir ProTide provided significantly enhanced carbovir triphosphate levels in HepG2 2.2.15 cells over that of the parent nucleoside. On the basis of these data, a series of phosphoramidate analogues with structural variation in the ester and amino acid regions were prepared and their antiviral profiles described. In addition, the pharmacokinetic disposition of the abacavir phenylethoxyalaninyl phosphoramidate was evaluated in Cynomolgus monkeys. PMID- 15887960 TI - Novel synthesis, cytotoxic evaluation, and structure-activity relationship studies of a series of alpha-alkylidene-gamma-lactones and lactams. AB - 5-Alkyl- and 5-arylalkyl-3-methylenedihydrofuran-2-ones 13a-e, 3 alkylidenedihydrofuran-2-ones 18a-c, and 3-methylenepyrrolidin-2-ones 16a-e were synthesized utilizing ethyl 2-diethoxyphosphoryl-4-nitroalkanoates 9a-e as common intermediates. All obtained compounds were tested against L-1210, HL-60, and NALM 6 leukemia cells. The highest cytotoxic activity was observed for 3 methylenefuranones 13d,e bearing benzyl or 3,4-dimethoxyphenylmethyl substituents at position 5, with IC(50) values of 5.4 and 6.0 microM, respectively. Contrary to the literature reports, no enhancement in activity due to the presence of a hydroxy group was found when the cytotoxicity of furanones 13a,b,d and 5-(1' hydroxyalkyl)-3-methylenedihydrofuran-2-ones 6a,b,d was compared. The anticancer activity of pyrrolidinones 16a-e and 3-alkylidenefuranones 18a-c was much weaker than that of furanones 13a-e. PMID- 15887961 TI - Design, synthesis, and antipicornavirus activity of 1-[5-(4-arylphenoxy)alkyl]-3 pyridin-4-ylimidazolidin-2-one derivatives. AB - A series of pyridylimidazolidinone derivatives was synthesized and tested in vitro against enterovirus 71 (EV71). On the basis of compound 33 (DBPR103), introduction of a methyl group at the 2- or 3-position of the linker between the imidazolidinone and the biphenyl resulted in markedly improved antiviral activity toward EV71 with IC(50) values of 5.0 nM (24b) and 9.3 nM (14a), respectively. Increasing the branched chain to propyl resulted in a progressive decrease in activity, while inserting different heteroatoms entirely rendered the compound only weakly active. The introduction of a bulky group (cyclohexyl, phenyl, or benzyl) led to loss of activity against EV71. The 4-chlorophenyl moiety in 14a was replaced with bioisosteric groups such as oxadiazole (28a-d) or tetrazole (32a,b), dramatically improving anti-EV71 activity and selectivity indices. Compounds 14a, 24b, 28b, 28d, and 32a exhibited a strong activity against lethal EV71, and no apparent cellular toxicity was observed. Three of the more potent imidazolidinone compounds, 14a, 28b, and 32b, were subjected to a large group of picornaviruses to determine their spectrum of antiviral activity. PMID- 15887962 TI - Early absorption and distribution analysis of antitumor and anti-AIDS drugs: lipid membrane and plasma protein interactions. AB - The interactions of a set of compounds of potential importance for anticancer and AIDS chemotherapy with lipid membranes and plasma proteins were studied with a surface plasmon resonance (SPR) based optical biosensor, giving valuable information on the absorption and distribution of the compounds. The technique allowed both effective screening of compounds and more detailed kinetic and mechanistic analysis of specific interactions. The interaction with two different types of lipid membranes could be reliably measured at a drug concentration as low as 20 microM, allowing analysis of poorly soluble compounds. Distribution was evaluated by investigation of the interactions with two human plasma proteins, human serum albumin (HSA) and alpha(1)-acid glycoprotein (AGP). Two apparent binding sites were clearly defined for HSA: one with rapid and one with slow association and dissociation rates. The sites appear to differ in accessibility and recognition characteristics rather than in their capacities to form strong complexes with drugs. PMID- 15887963 TI - AFMoC enhances predictivity of 3D QSAR: a case study with DOXP-reductoisomerase. AB - We present structure-activity relationships for 43 inhibitors of 1-deoxyxylulose 5-phosphate (DOXP)-reductoisomerase, derived from protein-based docking, ligand based 3D QSAR, and a combination of both approaches as realized by AFMoC (adaptation of fields for molecular comparison). DOXP-reductoisomerase (DXR) is a key enzyme of the non-mevalonate pathway for isoprenoid building blocks. This target has been characterized as having potential in the treatment of malaria with fosmidomycin, an established DXR inhibitor, presently in clinical trials. As part of an effort to optimize the properties of fosmidomycin, analogues have been synthesized and tested to gain further insights into the primary determinants of structural affinity. These data have been used to create a predictive model for DXR inhibition applying data taken from several DXR X-ray structures. These structures still leave the active fosmidomycin conformation and detailed reaction mechanism undetermined. This together with the small inhibitor data set provides a major challenge for presently available docking programs and 3D QSAR tools. To overcome these difficulties we have applied the AFMoC protocol. AFMoC makes more efficient use of available modeling data by tailoring DrugScore knowledge-based potentials specifically toward a given protein using inhibitor potency data. While 3D QSAR methods achieved valid models which lack predictivity, AFMoC was found to provide superior performance, based both on cross-validation runs as well as for inhibitors not considered in the training set. In particular, AFMoC's ability to gradually transform between generally applicable unadapted interaction fields to case specifically adapted ones proved to be of major importance. Using 50% tailored fields was found to permit the precise prediction of binding affinities for related ligands without losing the capability to estimate the affinities of structurally distinct inhibitors. PMID- 15887964 TI - Further studies on the interaction of the 5-hydroxytryptamine3 (5-HT3) receptor with arylpiperazine ligands. development of a new 5-HT3 receptor ligand showing potent acetylcholinesterase inhibitory properties. AB - Novel arylpiperazine derivatives bearing lipophilic probes were designed, synthesized, and evaluated for their potential ability to interact with the 5 hydroxytryptamine(3) (5-HT(3)) receptor. Most of the new compounds show subnanomolar 5-HT(3) receptor affinity. Ester 6bc showing a picomolar K(i) value is one of the most potent 5-HT(3) receptor ligands so far synthesized. The structure-affinity relationship study suggests the existence of a certain degree of conformational freedom of the amino acid residues interacting with the substituents in positions 3 and 4 of the quipazine quinoline nucleus. Thus, the tacrine-related heterobivalent ligand 6o was designed in an attempt to capitalize on the evidence of such a steric tolerance. Compound 6o shows a nanomolar potency for both the 5-HT(3) receptor and the human AChE and represents the first example of a rationally designed high-affinity 5-HT(3) receptor ligand showing nanomolar AChE inhibitory activity. Finally, the computational analysis performed on compound 6o allowed the rationalization of the structure-energy determinants for AChE versus BuChE selectivity and revealed the existence of a subsite at the boundary of the 5-HT(3) receptor extracellular domain, which could represent a "peripheral" site similar to that evidenced in the AChE gorge. PMID- 15887965 TI - Structure-based design: synthesis and biological evaluation of a series of novel cycloamide-derived HIV-1 protease inhibitors. AB - The structure-based design and synthesis of a series of novel nonpeptide HIV protease inhibitors are described. The inhibitors were designed based upon the X ray crystal structure of inhibitor 1 (UIC-94017)-bound HIV-1 protease. The inhibitors incorporated 3-hydroxysalicyclic acid-derived acyclic and cyclic P2 ligand into the (R)-(hydroxyethylamino)sulfonamide isostere. The inhibitors contain only two chiral centers and are readily synthesized in optically active form utilizing Sharpless asymmetric epoxidation, regioselective epoxide opening, and ring-closing olefin metathesis using Grubbs' catalyst as the key steps. We have synthesized 13-15-membered cycloamides and evaluated their HIV-1 protease enzyme inhibitory and antiviral activities in MT-2 cells. Interestingly, all cycloamide-derived inhibitors are noticeably more potent than the corresponding acyclic compounds. The ring size and substituent effects were investigated. It turned out that the 14-membered saturated ring is preferred by the S(1)-S(2) active sites of HIV-1 protease. Macrocycle 26 showed excellent enzyme inhibitory potency with a K(i) value of 0.7 nM and an antiviral IC(50) value of 0.3 microM. In view of their structural simplicity and preliminary interesting results, further optimization of these inhibitors is underway. PMID- 15887966 TI - Synthesis and structure-activity relationships of novel selective factor Xa inhibitors with a tetrahydroisoquinoline ring. AB - A series of novel 2,7-disubstituted tetrahydroisoquinoline derivatives were designed and synthesized. Among these derivatives, compounds 1 and 2 exhibited potent inhibitory activity against factor Xa (FXa) and good selectivity with respect to other serine proteases (thrombin, plasmin, and trypsin). In addition, compound 2 exhibited potent anti-FXa activity after intravenous and oral administration to cynomolgus monkeys, showed a dose-dependent antithrombotic effect at 0.1, 0.3, and 1 mg kg(-1) h(-1) in a rat model of venous thrombosis, and significantly reduced the size of brain infarction in a middle cerebral artery occlusion model at a dose of 0.1 mg kg(-1) h(-1). These results suggest that compound 2 (JTV-803) is likely to be useful as both a venous and arterial antithrombotic agent. PMID- 15887967 TI - Methyl substitution of 2-aminobicyclo[3.1.0]hexane 2,6-dicarboxylate (LY354740) determines functional activity at metabotropic glutamate receptors: identification of a subtype selective mGlu2 receptor agonist. AB - LY354740 (1) is a highly potent and selective agonist of metabotropic glutamate (mGlu) receptors 2 and 3. In the present study, we have prepared C3- and C4 methyl-substituted variants of rac-1, compounds 5, 9, and 13. Each of these racemic methyl-substituted analogues displaced specific binding of the mGlu2/3 receptor antagonist (3)H-2S-2-amino-2-(1S,2S-2-carboxycycloprop-1-yl)-3-(xanth-9 yl)propanoic acid ((3)H-LY341495) from membranes expressing mGlu2 or mGlu3 receptor subtypes. Evaluation of the functional effects of this series on second messenger responses in cells expressing human mGlu2 or mGlu3 receptors revealed C3beta-methyl analogue 5 to possess antagonist properties at both mGlu2 and mGlu3 receptors while C4beta-methyl analogue 9 acts as a full agonist at each of these targets. Unexpectedly, we found that incorporation of a methyl substituent at the C4alpha-position as in analogue 13 results in a mixed mGlu2 agonist/mGlu3 antagonist pharmacological profile. All of the mGlu2 agonist and mGlu3 antagonist activity of rac-13 was found to reside in its resolved (+)-isomer. PMID- 15887968 TI - Stereoselective binding of indomethacin ethanolamide derivatives to cyclooxygenase-1. AB - We have used molecular modeling studies and molecular dynamics simulations to generate three-dimensional models for cyclooxygenase-1 complexes with a series of indomethacin ethanolamide derivatives. These studies provide a plausible explanation for the stereoselective ligand binding preferences observed experimentally for these inhibitors and predict the general binding mode as well as specific structural details for the ligand-enzyme complexes. These studies provide insight into the nature of cyclooxygenase-1 interactions with a series of novel inhibitors and should help increase our understanding of key structural determinants for cyclooxygenase isozyme-selective inhibitor binding. PMID- 15887969 TI - Optimization of amide-based inhibitors of soluble epoxide hydrolase with improved water solubility. AB - Soluble epoxide hydrolase (sEH) plays an important role in the metabolism of endogenous chemical mediators involved in the regulation of blood pressure and inflammation. 1,3-Disubstituted ureas with a polar group located on the fifth atom from the carbonyl group of urea function are active inhibitors of sEH both in vitro and in vivo. However, their limited solubility in water and relatively high melting point lead to difficulties in formulating the compounds and poor in vivo efficacy. To improve these physical properties, the effect of structural modification of the urea pharmacophore on the inhibition potencies, water solubilities, octanol/water partition coefficients (log P), and melting points of a series of compounds was evaluated. For murine sEH, no loss of inhibition potency was observed when the urea pharmacophore was modified to an amide function, while for human sEH 2.5-fold decreased inhibition was obtained in the amide compounds. In addition, a NH group on the right side of carbonyl group of the amide pharmacophore substituted with an adamantyl group (such as compound 14) and a methylene carbon present between the adamantyl and amide groups were essential to produce potent inhibition of sEH. The resulting amide inhibitors have 10-30-fold better solubility and lower melting point than the corresponding urea compounds. These findings will facilitate synthesis of sEH inhibitors that are easier to formulate and more bioavailable. PMID- 15887970 TI - N-methylsansalvamide a peptide analogues. Potent new antitumor agents. AB - Sansalvamide A, a cyclic depsipeptide isolated from a marine fungus of the genus Fusarium, is composed of four hydrophobic amino acids (Phe, two Leu, Val) and one hydroxy acid ((S)-2-hydroxy-4-methylpentanoic acid; O-Leu) with five stereogenic centers all having S-stereochemistry. We have recently synthesized the corresponding cyclic peptide (Gu, W.; Liu, S.; Silverman, R. B. Organic Lett. 2002, 4, 4171-4174) and found that it too has antitumor activity. N-Methylation can enhance potency and selectivity for peptides. Consequently, here we synthesize 12 different N-methylated sansalvamide A peptide analogues and show that for several different tumor cell lines three of these analogues are more potent than the natural product; in pancreatic cells, sansalvamide A shows little activity, but the N-methylsansalvamide peptides are potent cytotoxic agents. PMID- 15887971 TI - Mono- and bis-thiazolium salts have potent antimalarial activity. AB - Three new series comprising 24 novel cationic choline analogues and consisting of mono- or bis (N or C-5-duplicated) thiazolium salts have been synthesized. Bis thiazolium salts showed potent antimalarial activity (much superior to monothiazoliums). Among them, bis-thiazolium salts 12 and 13 exhibited IC(50) values of 2.25 nM and 0.65 nM, respectively, against P. falciparum in vitro. These compounds also demonstrated good in vivo activity (ED(50) 18 years old) diabetics living in the Bella Coola Valley. Prevalence rates for history of alcohol issues, retinopathy, coronary artery disease, cerebrovascular disease, peripheral vascular disease, peripheral neuropathy, hypertension, hypercholesterolemia, and nephropathy were 44%, 14%, 19%, 8%, 7%, 10%, 54%, 47%, and 7% respectively. For the 1597 non-diabetics living in the Bella Coola Valley, respective prevalence rates for these same co morbidities were 20%, 0.3%, 2%, 1.5%, 1%, 1%, 10%, 6%, and 0.6%. The study did not demonstrate that Aboriginal people living in the Bella Coola Valley have an increased prevalence of diabetes associated co-morbidities over and above that found in the non-Aboriginal diabetic population. This was despite the fact the smoking rate was higher in the Aboriginal population. CONCLUSIONS: The development of diabetes in both Aboriginal and non-Aboriginal people living in the Bella Coola Valley was clearly associated with the presence of multiple co morbidities, including hypertension, hypercholesterolemia, coronary artery disease, cerebrovascular disease, and neuropathy. Rates of diabetes associated co morbidities were similar for both Aboriginal and non-Aboriginal diabetic populations. The authors speculate that a diet rich in fish oils (omega-3 fatty acids) accounted for the lower than expected rates of cardiovascular disease among this Aboriginal population. PMID- 15887993 TI - Can medical students learn specialist disciplines based in rural practice: lessons from students' self reported experience and competence. AB - INTRODUCTION: Medical schools in Australia are being funded by the Commonwealth Government Department of Health and Aging to move a considerable amount of undergraduate clinical education into rural and remote settings. There are concerns that these students may be disadvantaged in terms of exposure to appropriate clinical learning opportunities. This study compared learning opportunities for students undertaking an entire clinical year based in a rural primary care setting, a remote secondary hospital, or a traditional urban tertiary teaching hospital. METHODS: Twenty-nine students, six from rural primary care, eight from the remote secondary hospital, and 15 from the urban tertiary teaching hospital, completed a retrospective survey of their experience and perceived competence to manage 78 common procedural skills and 62 common conditions. RESULTS: Students in rural primary care reported a pattern of increased clinical exposure to common clinical conditions and procedures in comparison with their hospital-based peers. In comparing the two hospital-based programs, the students in the remote secondary care hospital reported increased exposure to common conditions and no significant difference in the opportunity to undertake common procedures. The data also demonstrated that there was a positive correlation between reported experience and self-perceived competence, and that this was greater for procedural skills than competence in managing common conditions. CONCLUSIONS: This study provides further evidence that rural primary care is an excellent setting for high quality clinical and educational experiences. These findings should serve to encourage students and staff involved with the new Rural Clinical School programs. PMID- 15887995 TI - 2004: The end of the beginning for Rural and Remote Health. PMID- 15887994 TI - Preliminary evidence from Queensland that rural clinical schools have a positive impact on rural intern choices. AB - INTRODUCTION: The unequal and inequitable distribution of the medical workforce between rural and urban parts of Australia has been well documented. Commonwealth and state governments have introduced several significant initiatives in an attempt to address this imbalance, including recruitment of many overseas trained doctors. One longer-term initiative is the funding of university departments of rural health and rural clinical schools in medical schools. OBJECTIVE: To determine the impact of the rural clinical division of the School of Medicine at the University of Queensland (UQ), Australia, on the intern workforce in central and southern Queensland, Australia. METHODS: Time series analysis of first preferences for intern allocation among UQ graduates and source of interns (UQ, interstate and overseas) from 2001-2005, and comparison of trends between Rockhampton and Toowoomba (UQ student placements since 2003) with Mackay (no placements). RESULTS: First preferences for Rockhampton increased from six in 2001 to 10 in 2005, and for Toowoomba from five in 2002 to 12 in 2005, while for Mackay preferences were stable at two. At Rockhampton while two interns came from overseas in 2001 and three were from interstate in 2002, UQ provided all interns in 2004 or 2005. UQ has provided 12/13 interns in 2004 and 13/14 in 2005 for Toowoomba. Mackay continues to source interns from interstate and overseas with UQ providing only 3/5 interns in 2004 and 2005. At Rockhampton, among non-bonded UQ graduates the number of interns choosing to work there increased from zero in 2001 to six in 2005. For Toowoomba, numbers were seven and 10 respectively, while for Mackay it was zero. CONCLUSIONS: UQ's rural clinical division is having a positive impact on the intern workforce in the regional hospitals most closely allied with it. PMID- 15888020 TI - Laparoscopic kidney donation: looking more than skin deep. PMID- 15888019 TI - Assessing renal function by GFR prediction equations in kidney transplantation. PMID- 15888021 TI - New insights into the pathogenesis and the therapy of recurrent focal glomerulosclerosis. AB - Recurrent focal glomerulosclerosis (FSGS) in renal allografts has remained a frustrating and enigmatic disease. Recent studies on gene mutations encoding podocin and other components of the slit-diaphragm in patients with native kidney nephrotic syndrome have underscored the heterogenecity of the idiopathic form of FSGS. While familial FSGS rarely recurs following transplantation, the sporadic variety of FSGS is associated with a 30% recurrence rate. The patients with the sporadic variety of FSGS who have homozygous or complex heterozygous podocin mutations have a low recurrence rate. In the other patients with sporadic FSGS, a more complex and likely multifactorial etiology accounts for the recurrence of FSGS. The role of CD80 expression on podocytes is intriguing but requires confirmation in kidney biopsies of patients with recurrent FSGS. Recent findings on podocin genomics, the permeability factor and CD80 expression may ultimately lead to a better understanding of recurrent FSGS as well as a more effective approach to its prevention and treatment. PMID- 15888022 TI - Effects of combined T- and B-cell deficiency on murine ischemia reperfusion injury. AB - B and T cells have been implicated in the pathogenesis of renal ischemia reperfusion injury (IRI); however, it is unknown if B and T cells interact in early injury responses, as seen in adaptive immune responses. Recent evidence has shown that B-cell deficient and T-cell deficient mice are partially protected from renal IRI. Renal IRI was induced in recombinase activating gene (RAG)-1 deficient mice, which lack both B and T cells. RAG-1 deficient mice from two different background strains were not protected from renal IRI. Adoptive transfer of either B or T cells into RAG-1 deficient mice led to a significant protection of renal injury, which was independent of effects on neutrophil trafficking. Neutrophil depletion in RAG-1 deficient mice did not protect from IRI. While deficiency of either B or T cells reduced IRI, combined lack of both is not protective. These results demonstrate that complex interactions between B and T cells are likely occurring in kidney IRI. PMID- 15888023 TI - Adenovirus-mediated bcl-2 gene transfer inhibits renal ischemia/reperfusion induced tubular oxidative stress and apoptosis. AB - Ischemia/reperfusion induces oxidative injury to proximal and distal renal tubular cells. We hypothesize that Bcl-2 protein augmentation with adenovirus vector mediated bcl-2 (Adv-bcl-2) gene transfer may improve ischemia/reperfusion induced renal proximal and distal tubular apoptosis through the mitochondrial control of Bax and cytochrome C translocation. Twenty-four hours of Adv-bcl-2 transfection to proximal and distal tubular cells in vitro upregulated Bcl-2/Bax ratio and inhibited hypoxia/reoxygenation induced cytochrome C translocation, O(2) (-) production and tubular apoptosis. Intra-renal arterial Adv-bcl-2 administration with renal venous clamping augmented Bcl-2 protein of rat kidney in vivo in a time-dependent manner. The maximal Bcl-2 protein expression appeared at 7 days after Adv-bcl-2 administration and the primary location of Bcl-2 augmentation was in proximal and distal tubules, but not in glomeruli. With a real-time monitoring O(2) (-) production and apoptosis analysis of rat kidneys, ischemia/reperfusion increased renal O(2) (-) level, potentiated proapoptotic mechanisms, including decrease in Bcl-2/Bax ratio, increases in caspase 3 expression and poly-(ADP-ribose)-polymerase fragments and subsequent proximal and distal tubular apoptosis. However, Adv-bcl-2 administration significantly enhanced Bcl-2/Bax ratio, decreased ischemia/reperfusion induced O(2) (-) amount, inhibited proximal and distal tubular apoptosis and improved renal function. Our results suggest that Adv-bcl-2 gene transfer significantly reduces ischemia/reperfusion induced oxidative injury in the kidney. PMID- 15888024 TI - Low-dose FK506 blocks collar-induced atherosclerotic plaque development and stabilizes plaques in ApoE-/- mice. AB - Since atherosclerosis is a chronic inflammatory disease, we tested the hypothesis that the immunosuppressive drug FK506 would attenuate the development of atherosclerosis using a mouse model of collar-induced atherosclerosis. ApoE-/- mice were treated for 4 weeks with the immunosuppressive drug FK506 (0.05 mg/kg/day), yielding sustained blood levels (approximately 0.2 ng/mL) without systemic side effects. Atherosclerotic plaque development of FK506-treated mice was significantly reduced (63%) while plaque cell density was increased (52%) compared to controls. Importantly, FK506 also blocked progression of pre-existing atherosclerotic plaques. Plaque area of pre-existing plaques was 35% reduced by FK506. Cell density (35%) and collagen content (51%) were significantly increased, whereas necrotic core content was decreased (42%), indicating a more stable plaque morphology. Similar results were found during spontaneous atherosclerotic plaque development in ApoE-/- mice (treatment 17-25 weeks of age). Flow-cytometric analysis showed no peripheral effects on blood cell count or T-cell activation after FK506-treatment. In vitro, FK506 decreased vascular smooth muscle cell (VSMC) apoptosis and inhibited nuclear factor of activated T cells (NFAT)-luciferase reporter activity at concentrations in the range of the in vivo concentration. Low-dose FK506 inhibits collar-induced atherosclerotic plaque development and progression and induces more stable plaque phenotypes in ApoE-/- mice without any peripheral side effects. PMID- 15888025 TI - In vivo molecular imaging characterizes pulmonary gene expression during experimental lung transplantation. AB - Experimental gene therapy is a promising strategy to prevent ischemia-reperfusion (I/R) injury and allograft rejection after lung transplantation, and methods will eventually be needed to characterize pulmonary transgene expression in vivo in humans. Therefore, we studied positron emission tomography (PET) as a means of performing in vivo molecular imaging in rodent models of lung transplantation. Rats were transfected endotracheally with adenovirus encoding a fusion gene of a mutant Herpes simplex virus-1 thymidine kinase and the green fluorescent protein gene (the former serving as an imaging reporter gene). Twenty-four hours after transfection, lungs were transplanted in groups representing normal transplantation, I/R injury and acute allograft rejection. Imaging was obtained either 24 h after transplantation to study reperfusion injury or 4 days after transplantation to study graft rejection. After imaging, lungs were excised and analyzed for thymidine kinase activity. Imaging detected transgene expression in transplanted lungs even in the presence of acute rejection or I/R injury. The PET imaging signal correlated with in vitro lung tissue assays of thymidine kinase activity (r(2) = 0.534). Thus, noninvasive molecular imaging with PET is a feasible, sensitive and quantitative method for characterizing pulmonary transgene expression in experimental lung transplantation. PMID- 15888026 TI - Recruitment of CXCR3+ and CCR5+ T cells and production of interferon-gamma inducible chemokines in rejecting human arteries. AB - Chemokine receptors preferentially expressed by Th1 cells and their IFN-gamma inducible ligands predominate in experimental and clinical allograft rejection. Previous chemokine-related transplantation studies have focused on parenchymal and microvascular inflammation which are of importance in acute rejection, but are not necessarily relevant in immune-mediated injury of conduit arteries. We have recently described a model of progressive human T cell-mediated infiltration and injury of allogeneic coronary artery segments using immunodeficient mouse hosts. In the present study, we investigated if recruitment of allogeneic T cells to different vascular compartments correlated with the expression of chemokines and their receptors. Transcripts were quantified by laser capture microdissection/real-time RT-PCR and their distribution was correlated to the corresponding protein expression detected by immunohistochemistry. Infiltrating T cells, confined to the adventitia and intima, expressed CXCR3 and CCR5, but were not recruited into the media despite production by vascular smooth muscle cells of IP-10, Mig, I-TAC, RANTES and MIP-1beta. Chemokine mRNA was detected primarily in vascular cells, although chemokine protein largely localized to infiltrating leukocytes which uniquely expressed their cognate receptors. These data explain the recruitment of IFN-gamma-secreting T cells to the vessel wall, and reinforce the suggestion that the arterial media may be a site of immunological privilege. PMID- 15888027 TI - The role of non-deletional tolerance mechanisms in a murine model of mixed chimerism with costimulation blockade. AB - Peripheral and central clonal deletion are important tolerance mechanisms in models using bone marrow transplantation (BMT) with costimulation blockade (CB). However, since tolerance can be found before peripheral deletion is complete and since elimination of recipient CD4(+) cells at the time of BMT prevents tolerance induction, we investigated the potential roles of regulation and anergy in such a murine model. We found that transient elimination of CD25(+) cells or neutralization of IL2 immediately after BMT and CB prevented the induction of skin graft tolerance. Cotransfer into SCID mice of CD4(+) cells taken from chimeras early after BMT, together with naive recipient-type CD4(+) cells significantly prolonged donor skin graft survival. In contrast, cotransfer of CD4(+) cells harvested from chimeras late after BMT did not prolong donor skin graft survival. Besides, depletion of CD25(+) cells in established chimeras several months post-BMT did not break tolerance. In vivo administration of recombinant IL2 inhibited chimerism and tolerance neither early nor late post BMT, arguing against a decisive role for classical anergy. Thus, CD4 cell mediated regulation contributes significantly to tolerance induction early after BMT, but appears to have no critical role in the maintenance of tolerance. PMID- 15888028 TI - Expression of human ecto-5'-nucleotidase in pig endothelium increases adenosine production and protects from NK cell-mediated lysis. AB - Ecto-5'-nucleotidase (E5'N) is an endothelial surface enzyme that controls conversion of extracellular nucleotides into immunosuppressive adenosine. We evaluated whether expression of human E5'N on pig endothelial cells (EC) attenuates human NK cell-mediated cytotoxicity. A pig EC line was stably transfected with human E5'N and human NK cell adhesion and cytotoxicity toward pig EC cultures was measured by flow cytometry and intracellular enzyme release. E5'N activity in pig EC lysates increased from 0.68 +/- 0.07 to 1013 +/- 293 nmol/min/mg protein, whilst the rate of AMP to adenosine metabolism by intact cells increased from 0.37 +/- 0.05 to >300 nmol/min/mg protein in non-transfected and transfected cells, respectively. The rate of adenosine production in transfected cells increased also with ATP as the extracellular substrate. Cytotoxicity of human NK cells was reduced from 10.7 +/- 0.4% and 11.1 +/- 1.1% with non-transfected pig EC to 5.2 +/- 0.2% and 5.0 +/- 0.2% in transfected cells with 50 microM and 250 microM AMP, respectively. Reduction of cytotoxicity in E5'N-transfected EC was abolished by the E5'N inhibitor and was mimicked in non transfected EC by the addition of adenosine, demonstrating the key role of adenosine produced by E5'N in inhibiting NK cell cytotoxicity. We suggest that overexpression of E5'N in EC of transgenic pigs is a possible strategy to ameliorate rejection after xenotransplantation. PMID- 15888029 TI - Pirfenidone inhibits lung allograft fibrosis through L-arginine-arginase pathway. AB - Transplant-related lung fibrosis is characterized by excessive fibro-collagenous deposition. Induction of arginase, an enzyme that metabolizes L-arginine to urea and L-ornithine, is vital for collagen synthesis. Pirfenidone is an investigational anti-fibrotic agent shown to be effective in blocking pulmonary fibrosis. The purpose of this study was to determine if pirfenidone was protective against the development of fibro-collagenous injury in rat lung orthotopic transplants through altering L-arginine-arginase metabolic pathways. Lung transplants were performed using Lewis donors and Sprague-Dawley recipients (allografts) or the same strain (isografts). Recipients were given pirfenidone (0.5% chow) 1-21-day post-transplantation. A significantly increased peak airway pressure (PawP) with excessive collagen deposition was found in untreated lung allografts. Pirfenidone treatment decreased PawP and collagen content in lung allografts. The beneficial effects were associated with downregulation of arginase protein expression and activity. In addition, pirfenidone decreased endogenous transforming growth factor (TGF)-beta level in lung allografts, and TGF-beta stimulated arginase activity in a dose-dependent manner in both lung tissue and fibroblasts. These results suggest that pirfenidone inhibits local arginase activity possibly through suppression of endogenous TGF-beta, hence, limiting the development of fibrosis in lung allografts. PMID- 15888030 TI - The influence of phosphatidylinositol 3-kinase/Akt pathway on the ischemic injury during rat liver graft preservation. AB - We aimed to investigate the role of phosphatidylinositol 3 (PI3)-kinase/Akt pathway on ischemic injury. Rat liver grafts were preserved in UW solution with different treatments and were compared by 1-week survival rates and morphological changes with those of the control group. PI3-kinase/Akt was significantly activated at the sites of Thr 308 and Ser 473 in the preserved grafts. Downstream target proteins, glycogen synthase kinase-3beta (GSK-3beta) and caspase-9, were inactivated. However, survival signal transduction from Akt to Bad was blocked by calcium release after activation of PI3-kinase/Akt. Significant activation of caspase-12, -3 and -7 contributed to cell apoptosis and severe ischemic injury was shown after 7 h of preservation by UW solution with insulin. Downregulation of phospho-Akt at Thr 308 and Ser 473 was due to partial inhibition of PI3 kinase/Akt pathway by LY294002. Activation of GSK-3beta and inactivation of caspase-12 and Bad could be found in the LY294002 groups in which the liver grafts showed less ischemic injury. Higher 1-week survival rates in the heparin, LY294002, and glucagon groups confirmed the dysregulation of the pathway. In conclusion, PI3-kinase/Akt pathway was dysregulated and contributed to ischemic injury during preservation. Heparin and LY294002 could improve graft viability by maintaining calcium homeostasis during preservation. PMID- 15888031 TI - Acute rejection after rat renal transplantation leads to downregulation of NA+ and water channels in the collecting duct. AB - Renal transplantation is associated with alterations of tubular functions and of the renin-angiotensin-aldosterone system. The underlying cellular and molecular mechanisms are unclear. We used an allogeneic rat renal transplantation model of acute rejection with and without immunosuppression by cyclosporine A (CsA) and a syngeneic model as control. Uninephrectomized Lewis or Lewis-Brown-Norway (LBN) rats received a kidney from LBN-rats. Renal transporters and receptors were analyzed by immunohistochemistry, semiquantitative RT-PCR and Western-blot analysis. Intracellular Na(+) was analyzed microfluorimetrically in isolated cortical collecting ducts. mRNA expression and function of the epithelial Na(+) channel (ENaC) and mRNA and protein expression of the water-channel AQP2 were downregulated in transplanted kidneys undergoing rejection. Expression of the serum- and glucocorticoid-kinase (Sgk1) was decreased and that of the ubiquitin protein ligase Nedd4-2 was increased. These changes were absent under CsA-therapy and in syngeneic model. Expression and function of the Na(+)-K(+)-ATPase, expression of the secretory K(+)-channel and of the mineralocorticoid receptor remained unchanged. Reduced ENaC function is likely due to decreased Sgk1- and increased Nedd4-2 mRNA expression leading to reduced ENaC expression in the membrane. These acute downregulations of ENaC and AQP2 may be triggered to reduce energy consumption in the distal nephron to protect the kidney immediately after transplantation. PMID- 15888032 TI - Progression of alloresponse and tissue-specific immunity during graft coronary artery disease. AB - Chronic rejection remains the major obstacle for long-term transplant survival. Both indirect alloresponse and tissue-specific autoimmunity have been implicated in its pathogenesis. The interrelationship between these two types of host anti graft response remains poorly understood. We have developed an immunosuppression free mouse model of graft coronary artery disease (GCAD), in which all FVB (H 2(q)) cardiac allografts placed into minor Ag (mHC)-mismatched DBA/1 (H-2(q)) hosts survived more than 112 days, and developed GCAD. We then examined the kinetics of both anti-mHC alloresponse and host autoimmunity against heart specific antigen, cardiac myosin (CM). At 8 days post-transplantation, recipient mice showed minimal intragraft inflammation and apoptosis, and limited expansion of allo-specific T cells. In addition, we observed early production of anti myosin IgG1 autoantibodies, which occurred in the absence of activated CM specific T lymphocytes. By day 56, GCAD indices, the numbers of mHC- and CM reactive T cells, and the levels of circulating allo- and CM-specific antibodies were all significantly increased. While host alloresponse was exhausted at 112 days post-transplant, T-cell reactivity against CM persisted. Our data suggest that both allo- and tissue-specific immunity might contribute to the induction of GCAD. They indicate that continual autoimmune response against graft tissue antigens may provide for GCAD sustenance. PMID- 15888033 TI - XIAP overexpression in islet beta-cells enhances engraftment and minimizes hypoxia-reperfusion injury. AB - Recent advances in clinical islet transplantation have allowed patients with type 1 diabetes to become insulin independent, but the procedure is limited since islets from two donors per recipient are typically required. This limitation arises because within a few days of the islets being embolized into the portal circulation, at least half of the transplanted beta-cells have undergone apoptotic cell death triggered by hypoxic and chemokine/cytokine-mediated stress. We hypothesized that the survival of beta-cells in the early post-transplant period would be enhanced if naturally occurring inhibitor of apoptosis proteins (IAPs) were transiently overexpressed in the grafts. In the present study, we used a growth-regulatable beta-cell line (betaTC-Tet) as a model for beta-cells within islets, and examined whether adenovirally delivered XIAP (X-linked IAP-a highly potent IAP) could enhance beta-cell survival. In vitro, XIAP-expressing betaTC-Tet cells were markedly resistant to apoptosis in an ischemia-reperfusion injury model system and following exposure to cytokines. When Ad-XIAP transduced betaTC-Tet cells were transplanted subcutaneously into immunodeficient mice, the grafts were able to reverse diabetes in 3 days, vs. 21 days for Ad-betaGal transduced cells. This approach may allow more efficient use of the limited existing supply of human islets. PMID- 15888034 TI - MDRD equations for estimation of GFR in renal transplant recipients. AB - After renal transplantation monitoring and detection of slight-to-moderate changes in GFR is a prerequisite for an optimal patient management. Recently, several equations to estimate GFR were developed and verified in the MDRD study cohort. However, little is known about the application of the MDRD formulas in the setting of renal transplantation. We prospectively conducted a study of the GFR estimates of the Cockcroft and Gault (C&G), MDRD6-, MDRD7 and the abbreviated MDRD (aMDRD) with the true GFR as measured by (99m)Tc-DTPA clearance in 95 consecutive patients 6.5, 5.3-7.7 years (mean, 95% CI) after renal transplantation. On average the DTPA clearance was 37.4, 34.4-40.4 mL/min/1.73 m(2), which differed significantly from estimates of GFR by C&G (52.6, 48.3-56.9 mL/min/1.73 m(2)), MDRD7 (44.8, 40.7-49.0 mL/min/1.73 m(2)), MDRD6 (43.8, 39.9 47.7 mL/min/1.73 m(2)) and aMDRD (46.6, 42.4-50.9 mL/min/1.73 m(2)). Bias was lowest for MDRD6 (6.4 mL/min/1.73 m(2)) and highest for C&G (15.2 mL/min/1.73 m(2)). Precision was similar for MDRD7 and aMDRD (10.6 and 11.1 mL/min/1.73 m(2)) but significantly better for MDRD6 (8.6 mL/min/1.73 m(2); p < 0.035). Accuracy within 50% of real GFR was 55.8% for C&G, 83.2% for aMDRD, 87.4% for MDRD7 and 90.5% for MDRD6. MDRD equations perform significantly better than the commonly used C&G formula. Moreover, the MDRD6 equation provides the best diagnostic performance, and should therefore be preferred in renal transplant recipients. PMID- 15888035 TI - Laparoscopic procurement of kidneys with multiple renal arteries is associated with increased ureteral complications in the recipient. AB - This study investigates the effect of renal artery multiplicity on donor and recipient outcomes after laparoscopic donor nephrectomy. Three-hundred and sixty one sequential procedures were performed over a 4-year period. Forty-nine involved accessory renal arteries; of these, 36 required revascularization and 13 were small polar vessels and ligated. The 312 remaining kidneys with single arteries served as controls. Study variables included operative times, blood loss, hospital stay, graft function and donor and recipient complications. Kidneys with multiple revascularized arteries had a longer mean warm ischemia time (35.3 vs. 29.2 min, p = 0.0003), and more ureteral complications (6/36 vs. 10/312, p = 0.0013) than single-artery controls. In contrast, ligation of a small superior accessory artery had no significant effect on donor operative time, blood loss, or complication rate while providing similar recipient graft function compared to single-artery controls. Renal artery number is important in selecting the appropriate kidney for laparoscopic procurement. Given the current excellent results with right-sided donor nephrectomy, kidneys with single arteries should be preferentially procured, irrespective of side. PMID- 15888036 TI - Determinants of transplant surgeons' willingness to provide organs to patients infected with HBV, HCV or HIV. AB - The common provision of organs to patients infected with hepatitis B virus (HBV) and hepatitis C virus (HCV), but not to those infected with human immunodeficiency virus (HIV), has been attributed to perceived or real differences in transplantation efficacy among these populations. However, other explanations remain possible. We surveyed all active U.S. transplant surgeons to identify determinants of their views of the propriety of transplantation among HBV-, HCV-, and HIV-infected patients. The 347 surgeons (56.1%) returning completed questionnaires believed that HCV- and HIV-infected patients have similar post-transplant survival (p = 0.9), but that both groups fare worse than HBV-infected patients (p < 0.00001 for both comparisons). Most transplant surgeons considered HBV- and HCV-infected patients to be appropriate transplantation candidates (p = 1.0 for this comparison), whereas one-third considered HIV-infected patients to be appropriate candidates (p < 0.00001 when compared with HBV- or HCV-infected patients). That surgeons are generally willing to transplant HCV-infected patients but not HIV-infected patients, and yet believe these groups will have similar post-transplant survival, suggests that survival estimates alone do not explain surgeons' choices. HIV-infected patients should have equal access to organs unless or until evidence emerges that they fare substantially worse than other potential recipients. PMID- 15888037 TI - Attitudes toward transplantation in U.K. Muslim Indo-Asians in west London. AB - The worldwide shortage of organs for transplantation makes it important to understand why some oppose donation. Attitudes vary with religion and ethnicity. Accordingly, we undertook a qualitative study of the attitudes of 141 U.K. Muslim Indo-Asians to organ donation. Participants were observed, focus group discussions held and in-depth individual interviews conducted. We identified a high level of alienation from the health care system in general. With respect to organ donation in particular, its importance was generally discounted, often in deference to authority figures within the community who appeared negatively disposed. The culture-specific issues arguing against donation included a sense of the sacredness of the body, a fatalistic approach to illness, a belief that organs took on an independent role as 'witnesses' to an individual's life on Judgement Day and an anxiety that the donor would have no control of the probity of the recipient of an organ. We believe these data suggest a need to improve in a culturally sensitive fashion the provision of health information provided to this community. PMID- 15888038 TI - Steroid avoidance versus steroid withdrawal after simultaneous pancreas-kidney transplantation. AB - Two steroid-sparing immunosuppressive regimens were prospectively compared in recipients of simultaneous pancreas-kidney transplants, one did not include steroids at all and the other included steroids for the first 3 months following transplantation. All patients received rabbit anti-thymocyte globulin, mycophenolate mofetil (MMF) and cyclosporine. Fifty patients were randomised in an open-label, single center and prospective study. The incidence of biopsy proven acute rejection during the first 12 months after transplantation was the primary endpoint of the study. The incidence of biopsy-proven acute rejection was 4% in both groups. No statistically significant difference in patient (96 and 100%), kidney (96 and 100%) or pancreas (84 and 92%) survival was observed 1 year after transplantation in the steroid avoidance and steroid withdrawal groups, respectively. The total number of adverse events (including severe ones), length of hospitalization and infectious episodes did not differ between groups. Blood glucose and insulin levels, lipid profile and hemoglobin A1C levels did not differ statistically between the two groups. However, the 1-year serum creatinine level was significantly higher in the steroid avoidance group (132 vs. 114 micromol/L; p = 0.02). Steroid avoidance and steroid withdrawal 3 months after transplantation are safe and effective regimens for diabetic patients with pancreas-kidney transplants. PMID- 15888039 TI - Impact of right lobe with middle hepatic vein graft in living-donor liver transplantation. AB - Technical improvements in adult-to-adult living-donor liver transplantation (LDLT) have led to the use of right-lobe grafts to overcome the problems encountered with 'small-for-size grafts'. The major controversy remains that the venous drainage from anterior segment substantially depends on tributaries of the middle hepatic vein (MHV), and deprivation of such tributaries may critically influence the postoperative graft function. Right-lobe grafts with MHV could resolve the potential problem of congestion in anterior segment. From December 2000 to January 2004, we performed 217 right-lobe LDLTs for adult patients. Of these, 40 patients received a right lobe with MHV graft (18.4%). The overall cumulative 3-year graft survival rate of a right lobe with (n = 40) and without MHV (n = 177) was 86.2% and 74.8% (p = NS). The proximal side of the MHV and the drainage vein of segment IV to the MHV (the left medial superior vein) were preserved in 24 patients. All of them needed venous interposition graft for anastomosis. All patients had a patent right hepatic vein (RHV) and MHV anastomosis during the follow-up period. We adopted the right lobe with MHV graft in 40 LDLT cases. Vein graft is essential for safe MHV anastomosis in cases which preserve proximal side of the MHV. PMID- 15888040 TI - Alemtuzumab (CAMPATH 1H) induction therapy in cadaveric kidney transplantation- efficacy and safety at five years. AB - Alemtuzumab is a powerful lymphocyte depleting antibody currently being evaluated in solid organ transplantation. This paper describes 5-year results of a single center study of alemtuzumab as induction in renal transplantation. Thirty-three renal transplant recipients received 20 mg alemtuzumab on day 0 and 1, followed by half-dose cyclosporin monotherapy (trough concentration 75-125 ng/mL) from day 3. They were compared in a retrospective contemporaneous-controlled manner with 66 kidney transplant recipients transplanted in the same period and center who received conventional immunosuppression with cyclosporin, azathioprine and prednisolone. In the alemtuzumab group 12% of recipients died compared to 17% in the control group (p = 0.48); likewise graft loss was similar in both groups (21% vs. 26%, respectively, p = 0.58). Incidence of acute rejection was also comparable at 5 years (31.5% vs. 33.6%), although the pattern of rejection was different with 14% patients in the alemtuzumab group experiencing rejection over 1 year post-transplant compared to none in the control group. There was no significant difference between groups in terms of infection or serious adverse events. While acknowledging the limitations of a relatively small single-center study, results suggest that alemtuzumab induction allowed satisfactory long-term patient and graft survival equivalent to that seen with standard triple immunosuppression, while avoiding steroid therapy. PMID- 15888041 TI - Clinical significance of an early protocol biopsy in living-donor renal transplantation: ten-year experience at a single center. AB - We report here our 10-year experience of a biopsy performed at day 14 after transplantation in 304 patients with stable graft function. The factors that may have influenced subclinical rejection were analyzed according to histology. The incidence of subclinical rejection was 13.2%. Addition of mycophenolate mofetile (MMF) as a primary immunosuppressant significantly decreased the incidence of subclinical rejection compared with patients without such treatment (odds ratio, 0.23; p < 0.05). On the other hand, HLA-DR antigen mismatch (odds ratio, 2.39) and unrelated donor (odds ratio, 2.10) were also significantly associated with decreased subclinical rejection (p < 0.05). The incidence of acute rejection in patients with normal findings was lower than in those with borderline changes or subclinical rejection (0.23 +/- 0.05 vs. 0.48 +/- 0.07 and 0.60 +/- 0.11, respectively; p < 0.05). The graft survival rates in patients with subclinical rejection were lower than in patients with normal or borderline changes at 1 (88.4% vs. 97.9% and 99.1%; p < 0.05), 5 (77.8% vs. 96.2% and 95.9%; p < 0.05) and 10 (62.3% vs. 96.2% and 93.7%; p < 0.05) years. Thus, a protocol biopsy performed on day 14 after transplantation is useful for predicting graft survival. Triple therapy including MMF, related donor and HLA-DR antigen match are important factors for reducing subclinical rejection in living-donor renal transplantation. PMID- 15888042 TI - Association between mannose-binding lectin levels and graft survival in kidney transplantation. AB - The mannose-binding lectin (MBL) pathway of complement is activated by pattern recognition. Genetic MBL variants are frequent and associated with low MBL serum levels. Higher MBL levels may be associated with more complement-mediated damage resulting in inferior graft survival. Pre-transplant serum samples from 266 consecutive deceased donor kidney transplant recipients were analyzed for MBL concentration by ELISA. Subsequently the cohort was analyzed for transplant related outcome. There was no significant difference in incidence of delayed graft function in recipients with a low MBL level (< or =400 ng/mL) compared to those with a higher MBL level (>400 ng/mL) (37.1 vs. 34.9%). At 10 years, death censored graft survival was 89.9% in patients with an MBL level below 400 ng/mL compared with 78.8% at a higher MBL level (p < 0.02). Multivariate analysis including traditional risk factors for graft loss showed an independent risk of 2.7 (95% CI 1.2-6.3) for death-censored graft loss if pre-transplant MBL levels were above 400 ng/mL. This difference was almost entirely explained by rejection associated graft loss (2.4 vs. 12.4%, p < 0.01). Higher MBL levels seem to be associated with a more severe form of rejection leading to treatment failure and graft loss. If these data can be confirmed, pre-transplant MBL levels may provide additional information for risk stratification prior to kidney transplantation. PMID- 15888043 TI - Epithelial to mesenchymal transition during late deterioration of human kidney transplants: the role of tubular cells in fibrogenesis. AB - The hallmark of failing renal transplants is tubular atrophy and interstitial fibrosis (TA/IF). Injury to tubular epithelial cells (TEC) could contribute to fibrogenesis via epithelial-mesenchymal transition (EMT). We examined the features of EMT in renal transplants that developed TA/IF. Biopsies from 10 allograft kidneys with impaired function and TA/IF and 10 biopsies from transplants with stable function were compared to their implantation biopsies. Relative to implantation biopsies, TEC in TA/IF kidneys showed loss of epithelial markers (E-cadherin, cytokeratin) with altered distribution. Some TEC also showed new cytoplasmic expression of mesenchymal markers vimentin, S100A4, and alpha smooth muscle actin (alpha-SMA) and collagen synthesis marker heat shock protein (HSP-47), both in deteriorating and atrophic tubules. Double immunostaining showed coexpression of cytokeratin and vimentin, S100A4 and HSP-47, indicating intermediate stages of EMT in TA/IF. These changes were absent or much less in transplants with stable function. EMT features in the TA/IF group correlated with serum creatinine (vimentin, S100A4, HSP-47), history of T-cell-mediated rejection (cytokeratin, S100A4) and proteinuria (cytokeratin). These findings support a model in which the TEC damage induces loss of epithelial features and expression of fibroblast features, as a common pathway of deterioration by either immunologic or nonimmunologic processes. PMID- 15888044 TI - Increased expression of senescence-associated cell cycle inhibitor p16INK4a in deteriorating renal transplants and diseased native kidney. AB - Some features of kidney transplants with dysfunction overlap the lesions of aging, such as tubular atrophy and interstitial fibrosis (TA/IF) without major glomerular abnormalities. Somatic cell limitations could contribute to deterioration in aging and disease states. Since expression of p16(INK4a), a cell cycle inhibitor associated with somatic cell senescence in vitro, is induced in aged kidney, we studied whether kidneys with dysfunction and TA/IF manifested increased p16(INK4a) expression. We performed p16(INK4a) immunostaining on transplanted kidneys and native kidneys with chronic renal diseases. At implantation, transplants manifested little TA/IF, and nuclear p16(INK4a) immunostaining was consistent with age. However, transplants biopsied for abnormal function displaying TA/IF showed strong nuclear and cytoplasmic p16(INK4a) staining, beyond the amount predicted for age. Both atrophic and non atrophic nephrons displayed increased p16(INK4a), suggesting that it was not simply a feature of atrophy. Epithelial p16(INK4a) staining was not increased in transplants with good function, but was increased in diseased native kidneys. The finding of increased p16(INK4a) expression in renal transplants and diseased kidneys with TA/IF and impaired function supports the concept that some cell senescence changes that accompany aging are also induced by injury and disease stresses. Thus, aging, injury and disease may share common pathways involving somatic cell senescence. PMID- 15888045 TI - Cytochrome P450 3A4 and P-glycoprotein activity and assimilation of tacrolimus in transplant patients with persistent diarrhea. AB - Renal transplant recipients suffering from persistent diarrhea have been repeatedly reported to have increased tacrolimus (Tac) trough levels. This study aimed to explore this phenomenon in detail in 15 renal transplant recipients with diarrhea, whose immunosuppression consisted of corticosteroids, mofetil mycophenolate and Tac. Both hepatic and intestinal CYP3A4 and PGP activity, important determinants of Tac bioavailability, were assessed, together with global CYP activity and investigations for gastrointestinal infection, function and morphology. Global CYP, CYP3A4, PGP and trough/dose levels of Tac were compared with diarrhea-free controls. In addition, a pharmacokinetic study of Tac was performed in 11 patients affected by diarrhea versus 9 controls. As expected, diarrhea was associated with increased Tac trough levels. An even stronger, significant increase of dose-normalized Tac levels was observed between 90 and 360 min after Tac intake. Time to peak concentration and drug half-life, however, were not altered. In addition, a concomitant decrease (+/-50%) of intestinal PGP activity was noticed in patients with diarrhea. For global CYP, CYP3A4 and hepatic PGP activity no such differences were noted. This pattern was not influenced by the specific cause of diarrhea. These data strongly suggest that persistent diarrhea is associated with an increased oral bioavailability of Tac. PMID- 15888046 TI - BOOP is common in cardiac transplant recipients switched from a calcineurin inhibitor to sirolimus. AB - While bronchiolitis obliterans organizing pneumonia (BOOP) has been associated with the use of sirolimus (SIR), the incidence in a consecutive group of patients given SIR to replace a calcineurin-inhibitor (CI) is unknown. Twenty-nine consecutive cardiac transplant recipients were switched from a CI to SIR to ameliorate CI-associated nephropathy or coronary graft atherosclerosis. Seven patients (24%) developed BOOP. The clinical characteristics and biopsy results of these patients are presented. The clinical course and response to withdrawal of SIR in all and steroids in four of seven patients suggested the diagnosis of BOOP. Chest X-rays and CT scans showed typical findings of BOOP in all seven patients. Infection was excluded in all patients. Biopsy results were characteristic of BOOP in six of seven patients. Six patients recovered and one died. BOOP is a common and potentially serious adverse event in cardiac transplant patients switched from a CI to SIR, especially when SIR is started late post-transplantation. PMID- 15888047 TI - Effects of hemi-portocaval shunts for inflow modulation on the outcome of small for-size grafts in living donor liver transplantation. AB - Graft hyperperfusion in small-for-size grafts (SFSG) is considered the main causal factor of small-for-size syndrome (SFSS). We compared SFSG with a graft-to recipient body ratio < or =0.8, with and without graft inflow modulation (GIM) by means of a hemi-portocaval shunt (HPCS). Thirteen patients underwent adult-to adult living donor liver transplantation (AALDLT): G1, n = 5 [4 right livers (RL) and 1 left liver (LL)] without GIM, and G2, n = 8 (4 RL and 4 LL) with GIM. In G2 patients, portal vein flow (PVF) was significantly reduced by HPCS: 190 +/- 70 mL/min/100 g liver in G2 vs. 401 +/- 225 ml/min in G1 (p = 0.002). One- and 6 month post-transplantation graft volume/standard liver volume (GV/SLV) ratio was of 72% and 79.5% in G1; 80% and 101% in G2 (p = ns). SFSS was observed in three G1 recipients (who were retransplanted), but in none of the G2 patients. At 1 year, patient and graft survival was respectively of 40% and 20% in G1, 87.5% and 75% in G2 (p = 0.024 and 0.03). It is concluded that drastic reduction of PVF by means of HPCS improves overall patient and graft survival by averting the occurrence of SFSS. Graft inflow modulation through HPCS reduces the risk of complications when transplanting SFSG in adult recipients. PMID- 15888048 TI - Long-term deterioration of kidney allograft function. AB - Although long-term survival after kidney transplantation is critically dependent on maintaining stable allograft function, few studies have examined renal allograft function over time. Using pooled data from 10 278 consecutive transplants at five centers, we calculated slopes of estimated glomerular filtration rates (eGFR) measured after 1, 6 and 12 months in 9515, 8861 and 7359 patients surviving > or =1, > or =6 and > or =12 months, respectively. Slopes of eGFR progressively diminished for patients transplanted during 1984-1989, 1990 1993, 1994-1998 and 1999-2002 (analysis of variance p < 0.0001 and p = 0.1245 for slopes measured after 1 and 6 months, respectively). Slopes measured after 12 months were less in the most recent era: -2.2 +/- 7.2 mL/min/1.73 m(2)/year, -2.3 +/- 6.6 mL/min/1.73 m(2)/year, -2.4 +/- 7.4 mL/min/1.73 m(2)/year and -1.4 +/- 10.9 mL/min/1.73 m(2)/year, respectively, p = 0.0058. Slopes measured after 1, 6 and 12 months each were less for transplantations during 1999-2002, after adjusting for multiple transplantation characteristics (p < 0.0001). Similarly, in Cox proportional hazards analysis, the risk (95% CI) for a 25% reduction in eGFR was 0.92 (0.85-1.01), p = 0.0736 during 1990-1994; 0.94 (0.82-1.08), p = 0.4111 during 1995-1998 and 0.78 (0.64-0.95), p = 0.0110 during 1999-2002 (compared to 1984-1989). We conclude that the rate of decline in allograft function after kidney transplantation has improved, suggesting that stable, long term function may be achievable. PMID- 15888049 TI - Treatment of EBV-related post-renal transplant lymphoproliferative disease with a tailored regimen including EBV-specific T cells. AB - The treatment of EBV-associated post-transplant lymphoproliferative disease (PTLD) poses a considerable challenge. Efforts have been made to define regimens based on combination of the available therapeutic agents, chosen and tailored on a patient-by-patient basis, with the aim of augmenting event-free patient and graft survival. Recently, autologous EBV-specific cytotoxic T-lymphocytes (CTL) have proved effective in enhancing EBV-specific immune responses and reducing viral load in organ transplant recipients with active infection. We investigated the use of a tailored combined approach including autologous EBV-specific CTL for the treatment of EBV-related PTLD developing after pediatric kidney transplantation. Five patients with disseminated monoclonal (n = 3) or localized polyclonal (n = 2) PTLD unresponsive to reduction of immunosuppression were enrolled. The patients with disseminated PTLD received 4-5 courses of reduced dosage polychemotherapy, accompanied by rituximab on the first day of each course, while localized disease was removed surgically. At treatment completion, autologous EBV-specific CTL were infused. All patients showed a complete response to treatment, without therapy-related toxicity or rejection, and persist in remission with good renal function at a median follow-up of 31 months. These preliminary results suggest that a combined chemoimmunotherapy regimen including virus-specific T-cells is well tolerated and potentially effective as first-line treatment of EBV-related PTLD. PMID- 15888050 TI - Reduction of CMV disease with steroid-free immunosuppresssion in simultaneous pancreas-kidney transplant recipients. AB - The impact of a prednisone-free immunosuppressive regimen was evaluated in simultaneous pancreas-kidney (SPK) recipients. Patient and graft survivals, rejection rates and the incidence of CMV disease were determined. Two hundred consecutive SPK transplant recipients received tacrolimus-based immunosuppression with (n = 100) or without (n = 100) chronic prednisone therapy. Patients were induced with lymphocyte depleting antibodies or IL-2 receptor blockers and received prophylactic antiviral therapy. Patient and graft survivals and rejection rates were not statistically significantly different between treatment groups. Two-year cumulative incidence of CMV in recipients in the prednisone-free protocol was reduced (7.2% vs. 16%; p = 0.15). Considering only recipients at highest risk (D+/R- or D+R+), incidence of CMV disease in the prednisone-free group (n = 61) compared to the steroid-treated group (n = 48) was reduced from 36% to 18% (p < 0.05). Multivariate analysis confirmed the independent effect of prednisone treatment on the incidence of CMV (RR 2.3; p = 0.04). In the prednisone-free protocol, incidence of CMV was less frequent in recipients receiving induction with Campath versus rabbit antilymphocyte globulin (2.4% vs. 12.6%; p = 0.14). Eliminating prednisone immunotherapy did not adversely affect outcomes and was associated with a reduced rate of CMV in SPK recipients of organs from sero-positive donors. PMID- 15888051 TI - Intestinal transplantation under tacrolimus monotherapy after perioperative lymphoid depletion with rabbit anti-thymocyte globulin (thymoglobulin). AB - Modifications in the timing and dosage of immunosuppression can ameliorate the morbidity and mortality that has prevented widespread use of intestinal transplantation (ITx) in children. Thirty-six patients receiving ITx, aged 5 months to 20 years were given 2-3 mg(kg of rabbit anti-thymocyte globulin (rATG, thymoglobulin) just before ITx, and 2-3 mg(kg postoperatively (total 5 mg(kg). Twice daily doses of tacrolimus (TAC) were begun enterally within 24 h after graft reperfusion with reduction of dose quantity or frequency after 3 months. Prednisone or other agents were given to treat breakthrough rejection. After 8-28 months follow-up (mean 15.8 +/- 5.3), 1- and 2-year patient and graft survival is 100% and 94%, respectively. Despite a 44% incidence of acute rejection in the first month, 16 of the 34 (47%) survivors are on TAC (n = 14) or sirolimus (n = 2) monotherapy; 15 receive TAC plus low dose prednisone; one each receive TAC plus sirolimus, TAC plus azathioprine and TAC plus sirolimus and prednisone. There was a low incidence of immunosuppression-related complications. This strategy of immunosuppression minimized maintenance TAC exposure, facilitated the long-term control of rejection, decreased the incidence of opportunistic infections, and resulted in a high rate of patient and graft survival. PMID- 15888052 TI - Effectiveness of prophylactic Anti-HBV therapy in allogeneic hematopoietic stem cell transplantation with HBsAg positive donors. AB - Use of hepatitis B surface antigen (HBsAg) positive donors for allogeneic hematopoietic stem cell transplantation (HSCT) causes serious hepatitis B virus (HBV)-related liver morbidity and mortality in the recipient. We compared the effectiveness of anti-HBV therapy in 29 recipients who underwent HSCT using HBsAg positive marrow (group I) against a historical control group of 25 patients who received HBsAg positive marrow without pre-HSCT prophylaxis (group II). Anti-HBV therapy consisted of lamivudine for HBsAg-positive donors and all recipients (n = 29) as well as HBV vaccination to all HBsAg-negative recipients (n = 10) before HSCT. After transplantation, HBV-related hepatitis was significantly higher in group II than group I recipients [12 of 25 recipients (48%) vs. 2 of 29 recipients (6.9%), p = 0.002] and in recipients whose donors had detectable serum HBV DNA by Digene Hybrid Capture II assay [8 of 14 recipients (57.1%) vs. 6 of 40 recipients (15.0%), p = 0.02]. Six recipients in group II and none in group I died of HBV-related hepatic failure (24.0% vs. 0%, p = 0.01). By multivariate Cox analysis, anti-HBV therapy effectively reduces post-HSCT HBV-related hepatitis (p = 0.01, adjusted hazards ratio 7.27, 95%CI 1.62-32.58). Our data support the use of prophylactic therapy in preventing HBV-related hepatitis after allogeneic HSCT from HBsAg-positive donor. PMID- 15888053 TI - Everolimus alters the bronchoalveolar lavage and endobronchial biopsy immunologic profile post-human lung transplantation. AB - Everolimus has recently shown promise in terms of short- and long-term clinical lung transplant outcomes. This study aims to determine the altered lung allograft cellular and cytokine mileau when everolimus is substituted for azathioprine (AZA). Twenty-three stable lung transplantation (LTx) recipients were randomized in a double-blinded study to receive everolimus (13) or AZA (10) plus standard cyclosporine/prednisolone. Bronchoalveolar lavage (BAL) and endobronchial biopsies (EBB) were performed on three occasions (T(0)-T(2)) to elucidate cellular and cytokine profiles via immunocytochemistry, immunohistology and enzyme-linked immunosorbent assay (ELISA) techniques. There were no group differences for demographics or clinical events throughout the study nor baseline cellular/cytokine differences. BAL lymphocyte percentage fell in the AZA group by T(2) (p = 0.05). BAL and EBB CD4 measures significantly declined in the everolimus group by T(2) (p < 0.05). EBB neutrophils rose significantly in the AZA group, with a fall in the everolimus group resulting in a significant difference at T(2) (p = 0.01). In conclusion, everolimus has contributed to potentially important differences in BAL and EBB cellular profiles. PMID- 15888054 TI - Hepatitis C virus antibody status and survival after renal transplantation: meta analysis of observational studies. AB - The natural history of hepatitis C virus (HCV) among patients after renal transplantation (RT) remains incompletely defined. We conducted a systematic review of the published medical literature on the impact of hepatitis C antibody status on survival of patients who received RT. We used the random effects model of DerSimonian and Laird to generate a summary estimate of the relative risk (RR) for mortality and graft loss with HCV seropositivity across the published studies. We identified eight clinical trials (6365 unique patients); six (75%) were cohort studies and two (2/8 = 25%) controlled trials, respectively. Pooling of study results demonstrated that presence of anti-HCV antibody was an independent and significant risk factor for death and graft failure after RT; the summary estimate for RR was 1.79 (95% CI, 1.57-2.03; homogeneity test, p = 0.0427) and 1.56 (95% CI, 1.35-1.80; homogeneity test, p = 0.0192), respectively. As a cause of death, hepatocellular carcinoma (HCC) and liver cirrhosis were significantly more frequent among anti-HCV positive than anti-HCV negative RT patients. This meta-analysis demonstrates that RT recipients with anti-HCV antibody have an increased risk of mortality and graft failure compared with HCV antibody negative patients. PMID- 15888055 TI - A trial of valganciclovir prophylaxis for cytomegalovirus prevention in lung transplant recipients. AB - Cytomegalovirus (CMV) infection is common after lung transplantation. We performed a prospective trial of valganciclovir prophylaxis in lung recipients with outcomes compared to matched historical controls. The valganciclovir group (n = 40) (including D+/R- and R+ patients) was prospectively enrolled, and received oral valganciclovir 900 mg once daily for 12 weeks. Historical controls (n = 40) received 12 weeks of daily intravenous ganciclovir if D+/R- or 12 weeks of oral ganciclovir if R+. CMV viral load testing was done at two-week intervals until 6 months posttransplant. Baseline demographics and immunosuppression were comparable in the two groups. The incidence of CMV viremia was 16/40 (40.0%) in the valganciclovir arm versus 18/40 (45%) in the ganciclovir arm (p = NS). The incidence of symptomatic CMV disease was 8/40 (20%) versus 7/40 (17.5%), respectively (p = NS). In both groups viremia, while on prophylaxis, was uncommon (valganciclovir: 0/40 and ganciclovir: 2/40). Peak viral load and time to viremia were similar in the two arms. High rates of viremia and symptomatic disease occurred in the D+/R- patients after discontinuation of prophylaxis. Genotypic CMV sequence analysis demonstrated low rates of ganciclovir resistance in both groups. Valganciclovir prophylaxis had similar efficacy to either intravenous ganciclovir (D+/R- patients), or oral ganciclovir (R+ patients) in lung recipients. PMID- 15888056 TI - Acute renal failure after lung transplantation: incidence, predictors and impact on perioperative morbidity and mortality. AB - The incidence, predictors and clinical significance of acute renal failure (ARF) after lung transplantation are not well described. We retrospectively collected data on 296 patients transplanted at our center between April 1992 and December 2000; follow-up was extended until December 2002. Patients were initially divided into two groups: ARF (doubling of baseline creatinine within 2 weeks after surgery) and NoARF. The ARF group was subdivided into ARFD (dialyzed) and ARFnD (not dialyzed). The incidence of ARF was 56% (166/296), but most cases were ARFnD (n = 143). Independent predictors of ARFD (n = 23) were: baseline GFR (OR 0.98, CI 0.96-0.99, p = 0.012), pulmonary diagnosis other than COPD (OR 6.80, CI 1.5 30.89, p = 0.013), mechanical ventilation > 1 d (OR 6.16, CI 1.70-22.24, p = 0.006) and parenteral amphotericin B use (OR 3.04, CI 1.03-8.98, p = 0.045). Both ARFnD and ARFD were associated with longer duration of mechanical ventilation, increased hospital stay and increased early mortality. One-year patient survival was 92.3%, 81.8% and 21.7% in the NoARF, ARFnD and ARFD groups, respectively (p < 0.0001). After controlling for important covariates, ARFD remained associated with an increased hazard of dying (HR 6.77, CI 4.00-11.44, p < 0.0001). In conclusion, ARF occurs commonly after lung transplantation and affects important clinical outcomes, especially when dialysis is required. PMID- 15888057 TI - Tacrolimus pharmacokinetics and dose monitoring after lung transplantation for cystic fibrosis and other conditions. AB - In cystic fibrosis (CF), absorption of tacrolimus through the gastrointestinal tract may be impaired due to fat malabsorption. The aim of this pilot study was to compare tacrolimus pharmacokinetics and inter- and intrasubject variability of exposure in stable lung transplant recipients with and without CF, and to determine the best single-time predictors of exposure. The study included 11 lung transplant recipients with CF and 11 without CF who received tacrolimus twice daily. Blood samples were obtained predose and at 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 8 and 12 h postdose on 3 separate days within 1 week. Tacrolimus pharmacokinetics and inter- and intrasubject variability of exposure were similar in the two groups, though exposure-per-milligram-dose was approximately 50% lower in CF patients. Tacrolimus trough concentration did not accurately predict the area under the concentration curve (AUC(0-12)), but the concentration measured 3 h postdose (C(3)) was tightly correlated with the AUC(0-12) in both CF (r(2)= 0.86) and non-CF (r(2)= 0.92) patients. In summary, patients with CF have a higher tacrolimus oral clearance, but nonsignificant differences in short-term inter- and intrasubject variability of exposure compared to patients without CF. C(3) is tightly correlated with AUC(0-12) in lung transplant recipients with and without CF. PMID- 15888058 TI - Differences in CMV-specific T-cell levels and long-term susceptibility to CMV infection after kidney, heart and lung transplantation. AB - Patients after kidney, heart and lung transplantation differ in their immunosuppressive drug regimens and in susceptibility to infectious complications with cytomegalovirus (CMV). In this study, CMV-specific T-cell responses were characterized in long-term transplant recipients and associated with the frequency of infectious complications. CMV-reactive CD4 T cells from 50 healthy controls, 68 renal, 14 heart and 24 lung transplant recipients were flow cytometrically quantified by the induction of cytokines after specific stimulation. Moreover, the immunosuppressive effect of calcineurin inhibitors on specific T-cell reactivity was quantified in vitro and compared with responses in vivo. Median CMV-specific T-cell frequencies in long-term renal (1.48%; range 0.06-17.26%) and heart transplant recipients (0.90%; 0.13-12.49%) did not differ from controls (1.82%; 0.26-21.00%). In contrast, CMV-specific T-cell levels were significantly lower in lung transplant recipients (0.50%; <0.05-4.98%) and showed a significant correlation with the frequency of infectious episodes (r =-0.57, p = 0.005). The differences within the groups were associated with increasing dosages of immunosuppressive drugs, as exemplified for calcineurin inhibitors that dose dependently reduced specific T-cell reactivity in vitro. In conclusion, monitoring CMV-specific CD4 T cells may serve as a measure for long-term disease susceptibility and may contribute to an improved management of CMV complications after lung transplantation. PMID- 15888059 TI - Local complement C3 expression is upregulated in humoral and cellular rejection of renal allografts. AB - Evidence on the role of the complement system in transplantation pathology has been accelerated by the discovery of C4d as an in situ marker of antibody mediated rejection. However, a local or systemic source of complement expression during acute rejection is under discussion. Thus, we quantitatively analyzed local RNA expression of complement component C3 as a pivotal molecule in active humoral and cellular rejection of renal allografts. After laser microdissection, real-time RT-PCR was performed for C3 using RNA extracted from tubuli and glomeruli of 68 paraffin-embedded renal allograft biopsies. Protocol and indication biopsies with signs of humoral and/or cellular rejection were investigated. Quantitative expression analysis of cytokines (IFN gamma, MCP-1, IL2, IL8) potentially influencing local C3 expression was performed. We observed a significant increase in median expression level of C3 mRNA in tubuli of C4d positive indication biopsies, and in tubuli from indication biopsies with signs of T-cell-mediated cellular rejection. Highest expression levels were found in C4d-positive indication biopsies with signs of cellular rejection. Biopsies with upregulated C3 showed increased IFN gamma expression, suggesting allograft infiltrating T-cells as potential stimulus for local C3 expression. Therefore, locally synthesized complement component C3 contributes to both humoral and cellular rejection, with tubular epithelial cells being a major source. PMID- 15888060 TI - Chimerism in kidneys, livers and hearts of normal women: implications for transplantation studies. AB - Tissue chimerism was recently described in transplanted organs from female donors into male recipients, by demonstration of the Y-chromosome in tissue-derived cells. It was claimed that these Y-chromosome positive cells were recipient derived. To find out whether the chimeric cells, derived from pregnancies of sons or blood transfusions, could have been present in the solid organs before transplantation, we performed the following study. In situ hybridization for the Y-chromosome was performed on the normal organs (51 kidneys, 51 livers, 69 hearts) from 75 women of the normal population, whose child and blood transfusion status were known. Chimeric cells were found in 13 kidneys, 10 livers and 4 hearts, of 23 women. There was no relation between the child status or the blood transfusion history with the presence of Y-chromosome positive cells. We have for the first time demonstrated that male cells are present in normal kidneys, livers and hearts. Theoretically, these organs could have been used for the transplantation. Therefore, our findings demonstrate that the chimeric cells thus far described in transplantation studies, are not necessarily donor derived, and could have been present in the organs before the transplantation. PMID- 15888061 TI - Preferences, knowledge, communication and patient-physician discussion of living kidney transplantation in African American families. AB - It is unknown whether patient-physician discussion about live kidney transplantation (LT) among African Americans (AA) is affected by preferences, knowledge and family discussions regarding LT. We recruited 182 AA dialysis patients and their families and assessed the relation of preferences, knowledge and family discussions regarding LT to the occurrence of patient-physician discussion using multivariable logistic regression. Most patients (76%) desired a transplant, and most patients (93%), spouses (91%) and children (88%) had knowledge of LT. Nearly half of the families discussed transplantation. Only 68% of patients and less than half of their spouses (41%) and children (31%) had discussed transplantation with physicians. Patient-physician discussion was more common among patients: whose spouses acknowledged their interest in transplantation (adjusted odds ratio (AOR) (95% CI):3.5 (1.61-7.8)); who discussed transplantation with spouses (AOR(95% CI):5.25 (2.22-12.41)); whose spouses agreed that they discussed transplantation with patients (AOR (95%CI):5.20 (1.76-15.37)) and whose children discussed transplantation with patients' physicians (AOR (95%CI):7.4 (1.3-40.0)). Universal patient-physician discussion of LT does not occur despite patient preferences. Rates of family physician discussions are low, and rates of family discussions vary. Early family physician discussion of LT, use of allied health professionals to promote family discussion of LT and the institution of culturally appropriate programs to enhance discussion of LT in AA families could improve rates of discussion and enhance patients' access to LT. PMID- 15888062 TI - En-bloc kidney transplantation in the United states: an analysis of united network of organ sharing (UNOS) data from 1987 to 2003. AB - With increasing donor organ shortages, en-bloc kidney (EBK) transplantation is an alternative to utilize very young or very old donor age cadaver kidneys for transplantation. Several single-center series have reported excellent graft survival (GS). We sought to determine national level registry-based patterns for GS and determine adjusted hazard ratios (AHR) for graft loss after EBK versus single kidney (SK) cadaver transplants. Data reported to UNOS from 1987 to 2003 were analyzed using PHREG (SAS version 8.1) statistical procedures. Proportional hazards models were constructed that included multiple donor, recipient and surgical variables. Of the 2160 EBK transplants reported, 77% were from donors < 5 years of age. EBK transplants had superior GS to SK transplants, when donor age was restricted to < 5 years (AHR 0.708, p < 0.001). GS at 1, 3 and 5 years post transplant was superior with EBK (85%, 76% and 71%) versus SK (81%, 68%, 63% and p < 0.001 at all time points). EBK transplants from very young donors were associated with a significantly lower rate of delayed graft function than SK transplants (17.9% versus 23.4%, p < 0.001). National registry data suggest that EBK transplants present a viable option for transplantation of very young donor kidneys. PMID- 15888063 TI - Pre-donation assessment of kidneys by magnetic resonance angiography and venography: accuracy and impact on outcomes. AB - Reports on the accuracy of magnetic resonance angiography (MRA) and magnetic resonance venography (MRV) in evaluating living donor renovasculature employ few patients or omit the consequences of inaccurate scans. We retrospectively compared intraoperative findings to MRA/MRV scans in 146 donor-recipient pairs. For detecting accessory arteries and early branching, MRA sensitivity was 57.6%, specificity 96.5%, false positive rate 3.5%, false negative rate 42.4%, positive predictive value 82.6%, negative predictive value 88.6% and overall accuracy 87.7%. By excluding clinically inconsequential accessory arteries, MRA sensitivity rose to 73.1%, specificity to 96.7% and overall accuracy to 92.5%. For MRVs, sensitivity was 56.2%, specificity 99%, false positive rate 1%, false negative rate 43.8%, positive predictive value 90%, negative predictive value 94.8% and accuracy 94.5%. Inaccurate scans were associated with prolonged donor and recipient operations and more frequently reconstructed arteries, but did not affect clinical outcomes. Because most missed accessory arteries are inconsequential, MRA is a useful, less invasive method for defining donor renovascular anatomy. PMID- 15888064 TI - A prospective randomized trial of steroid-free maintenance regimens in kidney transplant recipients--an interim analysis. AB - We compared three maintenance immunosuppressive regimens in a rapid discontinuation of prednisone protocol. From March 1, 2001, through December 31, 2003, 239 first and second kidney transplant recipients (166 LD; 73 DD) were randomized. All recipients were treated with Thymoglobulin; all received steroids intraoperatively and for 5 days postoperatively. Randomization was to cyclosporine-mycophenolate mofetil (n = 85); high-level tacrolimus (TAC) (8-12 ng/mL)-low-level sirolimus (SRL) (3-7 ng/mL) (n = 72); or low-level TAC (3-7 ng/mL)-high-level SRL (8-12 ng/mL) (n = 82). We found no difference at 24 months between groups in patient, graft, death-censored graft, or acute rejection-free graft survival, or in kidney function. Wound complications were more common in SRL-treated recipients (p = 0.02); we found no other differences between groups in complication rates. Our data suggest that excellent patient and graft survival and low rejection rates can be obtained using a variety of maintenance protocols without prednisone. PMID- 15888065 TI - Heparin-induced thrombocytopenia in a renal transplant recipient. AB - Heparin-induced thrombocytopenia (HIT) type II is an immunologically mediated reduction in platelets that increases the risk of arterial or venous thrombosis. It has been reported in up to 5% of patients receiving unfractionated heparin. Unlike other thrombocytopenic coagulopathies, HIT is associated with a high risk of thromboembolic events if not treated with an appropriate anticoagulant alternative. Diagnosis is dependent on assessment of platelet reduction, identification of previous heparin exposure, detection of thrombotic complications and evaluation of laboratory assays. HIT has been well described in surgical patient populations; however, the abdominal organ transplant population is an exception. HIT should be included in the differential diagnosis of patients presenting with thrombocytopenia after transplantation in order to prevent or treat thrombotic complications that can pose a risk to patient or graft survival. PMID- 15888066 TI - A tissue-engineered artificial bile duct grown to resemble the native bile duct. AB - The aim of this study was to fabricate an artificial bile duct for the development of a new treatment for biliary diseases. Eighteen hybrid pigs were implanted with a bile duct organoid unit (BDOU) made of a bioabsorbable polymer. Twelve of the transplanted BDOUs had been seeded with autologous bone marrow cells (BMCs) in advance. Six animals, the controls, were grafted with the scaffold alone with no BMCs seeded. The common bile duct was cut, the hepatic cut end of the native common bile duct was anastomosed to the BDOU and the other end was anastomosed to the duodenum. The controls underwent a similar operation. The neo-bile duct was removed at pre-determined time points and investigated histologically. All 18 recipient pigs survived until their sacrifice at 6 weeks, 10 weeks or 6 months. Histological examination revealed incomplete epithelialization of the neo-bile duct at 6 weeks and 10 weeks after transplantation. At 6 months, the organoid exhibited a morphology almost identical to that of the native common bile duct. No differences were found between the controls and BMC-seeded pigs. These results show that the artificial bile duct thus fabricated can serve as a substitute for the native bile duct. PMID- 15888067 TI - Broncho-alveolar lavage matrix metalloproteases as a sensitive measure of bronchiolitis obliterans. AB - Bronchiolitis obliterans (BO) is a survival-limiting factor in lung transplantation. There are no common BO markers in use. Since BO is associated with extracellular matrix remodeling, we asked whether matrix metalloproteases (MMPs) and their tissue inhibitors (TIMPs) could serve as BO markers. In 72 lung transplant patients (34 BO syndrome (BOS) 0, 15 BOS 0-p, and 23 BOS 1) serum and broncho-alveolar lavage (BAL) MMP and TIMP levels were examined by ELISA. The BAL cell counts were additionally analyzed. The serum MMP-2, MMP-8, MMP-9 and TIMP-2 levels were not different in all groups. In contrast, the BAL MMP-8, -9 and TIMP 1 levels were significantly elevated in BOS 0-p (p = 0.003; p = 0.007; p = 0.0003, respectively) and BOS 1 (p = 0.003; p = 0.001; p = 0.0004, respectively) as compared to BOS 0 patients. The BAL MMP-8, -9 and TIMP-1 levels were significant predictors of BOS 0-p (p = 0.01; p = 0.01; p = 0.01, respectively) and BOS-1 (p = 0.007; p = 0.01; p = 0.006, respectively) in receiver operating characteristic analysis. Except for BAL macrophages that were significantly decreased in BOS 0-p versus BOS 0 patients; other cell counts were not different between the groups. BAL MMP-8, -9 and TIMP-1 might be useful markers to detect BO in lung transplant patients. PMID- 15888068 TI - The economic implications of noninvasive molecular testing for cardiac allograft rejection. AB - Endomyocardial biopsy is the mainstay for monitoring cardiac allograft rejection. A noninvasive strategy--peripheral blood gene expression profiling of circulating leukocytes--is an alternative with proven benefits, but unclear economic implications. Financial data were obtained from five cardiac transplant centers. An economic evaluation was conducted to compare the costs of outpatient biopsy with those of a noninvasive approach to monitoring cardiac allograft rejection. Hospital outpatient biopsy costs averaged 3297 US dollars, excluding reimbursement for professional fees. Costs to Medicare and private payers averaged 3581 US dollars and 4140 US dollars, respectively. A noninvasive monitoring test can reduce biopsy utilization. The savings to health care payers in the United States can be conservatively estimated at approximately 12.0 million US dollars annually. Molecular testing using gene expression profiling of peripheral circulating leukocytes is a new technology that offers physicians a noninvasive, less expensive alternative to endomyocardial biopsy for monitoring allograft rejection in cardiac transplant patients. PMID- 15888069 TI - Extended survival by urgent liver retransplantation after using a first graft with metastasis from initially unrecognized donor sarcoma. AB - A 58-year-old man underwent orthotopic liver transplantation for polycystic liver disease. Shortly after the procedure, it was discovered that the donor harbored a sarcoma of the aortic arch that had metastasized to the spleen, and bilateral renal cell carcinomas. The two sole organ recipients, our liver recipient and a lung recipient at another institution, were both listed for urgent retransplantation, which they received from the same second donor. The liver explant contained metastatic sarcoma. Twenty-four months survival following lung retransplantation has been previously reported. We report the 76-month disease free survival in the liver recipient. PMID- 15888070 TI - BK virus associated nephropathy in native kidneys of a heart allograft recipient. AB - Polyomavirus-mediated nephropathy is an increasingly recognized complication in renal transplant recipients, but data on the status of viral activity in the native kidneys of non-renal solid organ recipients are limited. Thirteen native kidney biopsies of heart transplant recipients with significant renal impairment were evaluated for the evidence of polyomavirus reactivation by immunohistochemistry and PCR. One case of BK virus-mediated nephropathy in a cardiac transplant recipient exposed to high levels of immunosuppressive drugs was identified. Clinical and histopathological findings of this patient progressing to terminal renal failure are discussed in detail. In conclusion, polyomavirus reactivation in native kidneys of heart transplant recipients can cause significant renal impairment and should be considered in the differential diagnosis in this patient cohort. PMID- 15888071 TI - Campath-1H use in pediatric renal transplantation. AB - Campath-1H is a humanized, monoclonal antibody directed against CD52 determinants on the surface of human B- and T-cells and monocytes. Reports of Campath-1H use as induction in adult renal transplantation have been encouraging with low rejection rates and minimal adverse events. We report four high risk pediatric kidney transplant patients who received Campath-1H for unique indications with variable results. Children ranged in age from 20 months to 16 years. Immunosuppression regimens varied. Three of four patients experienced acute rejection, two of which were C4d positive. Serial flow cytometry was performed on all four patients. The patient who received only Campath-1H has an absolute lymphocyte count that remains less than 50% of baseline at 12-months post transplant. In addition, in this patient CD3, CD4, CD8 and CD20 remain less than 50% of baseline. From this initial experience using Campath-1H in pediatric renal transplantation we conclude that; (1) the use of Campath-1H does not prevent recurrence of FSGS, (2) as seen in adults, lack of calcineurin inhibition when using Campath-1H may increase the risk of antibody-mediated rejection and (3) prolonged lymphocyte depletion remains even after a single dose of Campath-1H in children. PMID- 15888072 TI - Clinical practice guidelines for managing dyslipidemias in kidney transplant patients: lessons to be learnt from the assessment of Lescol in renal transplantation (ALERT) trial. PMID- 15888073 TI - Clinical practice guidelines for managing dyslipidemias in kidney transplant patients. PMID- 15888074 TI - Response to 'viral hepatitis guidelines for transplant recipients'. PMID- 15888075 TI - Use of anti-HCV-positive donor organs. PMID- 15888077 TI - Abstracts of the American Transplant Congress 2005. Seattle, Washington, USA. PMID- 15888076 TI - Composite tissue allografts: a challenge for transplantologists. PMID- 15888078 TI - Caenorhabditis elegans as a model for innate immunity to pathogens. AB - The amenability of the nematode Caenorhabditis elegans for genetic analysis and other experimentation provides a powerful tool for studying host-pathogen interactions. Our current understanding of how C. elegans responds to pathogen challenges is in its infancy, but the discovery that the worm has inducible defence responses, which to some extent parallel those of other organisms, demonstrates the potential of this model organism for the study of innate immunity. Most progress in dissecting the C. elegans antimicrobial response has focused around signal transduction pathways and the expression of genes activated by the worm in response to microbial infections. PMID- 15888079 TI - Erythrocyte variants and the nature of their malaria protective effect. AB - The malaria threat to global health is exacerbated by widespread drug resistance in the Plasmodium parasite and its insect vector, and the lack of an efficacious vaccine. Infection with Plasmodium parasites can cause a wide spectrum of pathologies, from a transient mild form of anaemia to a severe and rapidly fatal cerebral disease. Epidemiological studies in humans and experiments in animal models have shown that genetic factors play a key role in the onset, progression, type of disease developed and ultimate outcome of malaria. The protective effect of polymorphic variants in erythrocyte-specific structural proteins or metabolic enzymes against the blood-stage of the disease is one of the clearest illustrations of this genetic modulation, and has suggested co-evolution of the Plasmodium parasite with its human host in areas of endemic disease. Here, we present a brief overview of erythrocyte polymorphisms with biological relevance to malaria pathogenesis, and current work on the mechanism(s) by which these mediate their protective effect. The recent addition of erythrocyte pyruvate kinase to this group of protective genes will also be discussed. PMID- 15888080 TI - Autophagy is an immediate macrophage response to Legionella pneumophila. AB - After ingestion by macrophages, Legionella pneumophila enter spacious vacuoles that are quickly enveloped by endoplasmic reticulum (ER), then slowly transferred to lysosomes. Here we demonstrate that the macrophage autophagy machinery recognizes the pathogen phagosome as cargo for lysosome delivery. The autophagy conjugation enzyme Atg7 immediately translocated to phagosomes harbouring virulent Legionella. Subsequently, Atg8, a second autophagy enzyme, and monodansyl-cadaverine (MDC), a dye that accumulates in acidic autophagosomes, decorated the pathogen vacuoles. The autophagy machinery responded to 10-30 kDa species released into culture supernatants by Type IV secretion-competent Legionella, as judged by the macrophages' processing of Atg8 and formation of vacuoles that sequentially acquired Atg7, Atg8 and MDC. When compared with autophagosomes stimulated by rapamycin, Legionella vacuoles acquired Atg7, Atg8 and MDC more slowly, and Atg8 processing was also delayed. Moreover, compared with autophagosomes of Legionella-permissive naip5 mutant A/J macrophages, those of resistant C57BL/6 J macrophages matured quickly, preventing efficient Legionella replication. Accordingly, we discuss a model in which macrophages elevate autophagy as a barrier to infection, a decision influenced by regulatory interactions between Naip proteins and caspases. PMID- 15888081 TI - Effects of the Escherichia coli toxin cytolysin A on mucosal immunostimulation via epithelial Ca2+ signalling and Toll-like receptor 4. AB - Epithelial cells are vital to sense the presence of bacteria, thereby initiating a proper innate immune response. This occurs via different mechanisms, e.g. recognition by pattern recognition receptors (TLR), or alteration of the cellular Ca2+ homeostasis. The Escherichia coli toxin cytolysin A (ClyA) is naturally delivered to target cells as active pore assemblies within outer membrane vesicles (OMVs), and we here investigate a possible role of ClyA-containing OMVs (ClyA+ (OMV)) for induction of proinflammatory responses via the above-mentioned mechanisms. We report that low, sublytic concentrations of ClyA+ (OMV) affect the Ca2+ homeostasis in epithelial cells by induction of slow, intracellular Ca2+ oscillations, while increased concentrations act cytolytically. Thus, ClyA belongs to the novel group of pore-forming toxins shown to elicit such biphasic responses. Ca2+ waves in the minute range have been shown to regulate gene transcription of, e.g. interleukin (IL)-6 and -8. While the periodicity of ClyA+ (OMV)-induced Ca2+ waves (22.9 +/- 0.9 min) fail to induce an IL-8 response, our data fit to the general concept of frequency-specific gene expression. Molecular investigations of the signal transduction pathway reveals that ClyA+ (OMV) utilize a different one as compared with those previously reported for other toxins causing Ca2+ waves. The ClyA protein per se and ClyA pore assemblies are non-immunogenic, while lipopolysaccharide present on the OMVs induces a TLR4 dependent proinflammatory response as expected. Additional membrane components of the OMV, e.g. OmpW, was also found to elicit proinflammatory responses that was independent of TLR4 and Ca2+ signalling. PMID- 15888082 TI - Engagement of Toll-like receptors by mycoplasmal superantigen: downregulation of TLR2 by MAM/TLR4 interaction. AB - Mycoplasma arthritidis mitogen (MAM) is a superantigen (SAg) from M. arthritidis, an agent of murine toxic shock syndrome and arthritis. We previously demonstrated that C3H/HeJ and C3H/HeSnJ mice that differ in expression of TLR4 differed in immune reactivity to MAM. We show here that MAM directly interacts with TLR2 and TLR4 by using monoclonal antibodies to TLR2 and TLR4 which inhibit cytokine responses of THP-1 cells to MAM. Also, using macrophages from C3H substrains and TLR2-deficient mice, we confirmed that both TLR2 and TLR4 are used by MAM. Levels of IL-6 in supernatants of MAM-challenged macrophages were higher in mice which expressed only TLR2, lesser with both TLR2 and TLR4, and absent in mice lacking both TLR2 and TLR4. In addition, expression of TLR2 and TLR4 was moderately upregulated in wild-type cells but cells lacking TLR4 showed a fivefold increase in TLR2 expression. Further, blockade of TLR4 on macrophages of C3H/HeN mice with antibody greatly increased expression of TLR2 and release of IL-12p40 in response to MAM. These results indicate that the SAg, MAM, interacts with both TLR2 and TLR4 and that TLR4 signalling might downregulate the MAM/TLR2 inflammatory response. PMID- 15888083 TI - Suppression of Drosophila cellular immunity by directed expression of the ExoS toxin GAP domain of Pseudomonas aeruginosa. AB - We show here that transgenic Drosophila can be used to decipher the effect of a bacterial toxin on innate immunity and demonstrate the contribution of blood cells in fly resistance to bacterial infection. ExoS is a Pseudomonas aeruginosa exotoxin directly translocated into the host cell cytoplasm through the type III secretion system found in many Gram-negative bacteria. It contains a N-terminal GTPase activating (GAP) domain that prevents cytoskeleton reorganization by Rho family of GTPases in cell culture. Directed expression of the ExoS GAP domain (ExoSGAP) during fly eye morphogenesis inhibited Rac1-, Cdc42- and Rho-dependent signalling, demonstrating for the first time its activity on RhoGTPases in a whole organism. We further showed that fly resistance to P. aeruginosa infections was altered when ExoSGAP was expressed either ubiquitously or in haemocytes, but not when expressed into the fat body, the major source of NF-(kappa)B-dependent anti-microbial peptide synthesis. Fly sensitivity to infection was also observed with Gram-positive Staphylococcus aureus strain and was associated to a reduced phagocytosis capacity of ExoSGAP-expressing haemocytes. Our results highlight the major contribution of cellular immunity during the first hours after Drosophila infection by P. aeruginosa, an opportunist pathogen affecting patients with pathologies associated to a reduced leukocyte number. PMID- 15888084 TI - Distinct transcriptional profiles characterize oral epithelium-microbiota interactions. AB - Transcriptional profiling, bioinformatics, statistical and ontology tools were used to uncover and dissect genes and pathways of human gingival epithelial cells that are modulated upon interaction with the periodontal pathogens Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Consistent with their biological and clinical differences, the common core transcriptional response of epithelial cells to both organisms was very limited, and organism-specific responses predominated. A large number of differentially regulated genes linked to the P53 apoptotic network were found with both organisms, which was consistent with the pro-apoptotic phenotype observed with A. actinomycetemcomitans and anti apoptotic phenotype of P. gingivalis. Furthermore, with A. actinomycetemcomitans, the induction of apoptosis did not appear to be Fas- or TNF(alpha)-mediated. Linkage of specific bacterial components to host pathways and networks provided additional insight into the pathogenic process. Comparison of the transcriptional responses of epithelial cells challenged with parental P. gingivalis or with a mutant of P. gingivalis deficient in production of major fimbriae, which are required for optimal invasion, showed major expression differences that reverberated throughout the host cell transcriptome. In contrast, gene ORF859 in A. actinomycetemcomitans, which may play a role in intracellular homeostasis, had a more subtle effect on the transcriptome. These studies help unravel the complex and dynamic interactions between host epithelial cells and endogenous bacteria that can cause opportunistic infections. PMID- 15888085 TI - Secretion of Cpn0796 from Chlamydia pneumoniae into the host cell cytoplasm by an autotransporter mechanism. AB - By comparison of proteome profiles of purified Chlamydia pneumoniae and whole lysates of C. pneumoniae infected HEp-2 cells, an N-terminal fragment of the previously uncharacterized chlamydial protein Cpn0796 was identified as a secreted protein. A 38 kDa cleavage product of Cpn0796 was present in infected cells, whereas only the 65 kDa full-length Cpn0796 could be detected in purified Chlamydia. Confocal immunofluorescence microscopy demonstrated that Cpn0796 was localized in the Chlamydia membrane in young inclusions. However, at 36 h post infection and later Cpn0796 was detected in the cytoplasm of C. pneumoniae infected HEp-2 and BHK cells. Furthermore, Cpn0796 was detected in the cytoplasm of infected cells in the lungs of C. pneumoniae infected C57Bl mice. When cleavage was inhibited, Cpn0796 was retained in the chlamydiae. We propose that Cpn0796 is an autotransporter the N-terminal of which is translocated to the host cell cytoplasm. This is the first example of secretion of a Chlamydia autotransporter passenger domain into the host cell cytoplasm. Cpn0796 is specific for C. pneumoniae, where five homologous proteins are encoded by clustered genes. None of these five proteins were found to be secreted. PMID- 15888086 TI - Dissemination of Toxoplasma gondii to immunoprivileged organs and role of Toll/interleukin-1 receptor signalling for host resistance assessed by in vivo bioluminescence imaging. AB - Toxoplasma gondii infection can lead to life-threatening systemic disease in the immunocompromised individual and in the developing fetus. Despite intensive investigation in animal models of toxoplasmosis, the processes leading to systemic dissemination remain poorly characterized. In the present study, in vivo bioluminescence imaging (BLI) was applied to the Toxoplasma mouse model to study the dynamics of infection in real time. Photon emission analyses revealed rapid dissemination of parasites in the organism and dissemination to immunoprivileged organs (brain, eyes and testes). Spatio-temporal analysis by BLI in individual mice showed that the virulent RH strain (type I) and the non-virulent ME49/PTG strain (type II) disseminate widely, but the virulent RH strain (type I) exhibits a more dramatic expansion of parasite biomass. Assessment by BLI of the Toll/interleukin-1 receptor (TIR) signalling pathway in host resistance to T. gondii revealed that signal transduction to the adaptor protein MyD88 is probably mediated by Toll-like receptor(s) rather than by IL-1R or IL-18R signalling. However, TLR1(-/-), TLR2(-/-), TLR4(-/-), TLR6(-/-) and TLR9(-/-) animals did not exhibit increased susceptibility to infection. These results suggest that intricate mechanisms regulate TIR-mediated responses during Toxoplasma infection. PMID- 15888087 TI - Host cell lipids control cholesteryl ester synthesis and storage in intracellular Toxoplasma. AB - The intracellular protozoan Toxoplasma gondii lacks a de novo mechanism for cholesterol synthesis and therefore must scavenge this essential lipid from the host environment. In this study, we demonstrated that T. gondii diverts cholesterol from low-density lipoproteins for cholesteryl ester synthesis and storage in lipid bodies. We identified and characterized two isoforms of acyl CoA:cholesterol acyltransferase (ACAT)-related enzymes, designated TgACAT1alpha and TgACAT1beta in T. gondii. Both proteins are coexpressed in the parasite, localized to the endoplasmic reticulum and participate in cholesteryl ester synthesis. In contrast to mammalian ACAT, TgACAT1alpha and TgACAT1beta preferentially incorporate palmitate into cholesteryl esters and present a broad sterol substrate affinity. Mammalian ACAT-deficient cells transfected with either TgACAT1alpha or TgACAT1beta are restored in their capability of cholesterol esterification. TgACAT1alpha produces steryl esters and forms lipid bodies after transformation in a Saccharomyces cerevisiae mutant strain lacking neutral lipids. In addition to their role as ACAT substrates, host fatty acids and low density lipoproteins directly serve as Toxoplasma ACAT activators by stimulating cholesteryl ester synthesis and lipid droplet biogenesis. Free fatty acids significantly increase TgACAT1alpha mRNA levels. Selected cholesterol esterification inhibitors impair parasite growth by rapid disruption of plasma membrane. Altogether, these studies indicate that host lipids govern neutral lipid synthesis in Toxoplasma and that interference with mechanisms of host lipid storage is detrimental to parasite survival in mammalian cells. PMID- 15888088 TI - Lipooligosaccharide-independent alteration of cellular homeostasis in Neisseria meningitidis-infected epithelial cells. AB - Neisseria meningitidis (MC) is an important cause of meningitis and septic shock. Primary loose attachment of MC to host epithelial cells is mediated by type IV pili. Lipooligosaccharide (LOS), opacity (Opa) proteins and glycolipid adhesins facilitate subsequent tight attachment. MC infection causes numerous changes in host epithelial cell homeostasis. These include cortical plaque formation, increased expression of proinflammatory cytokines and alterations in host iron homeostasis. Using both biochemical and genetic approaches, we examined the role of LOS in mediating these events. We first examined specific cellular iron homeostasis changes that occur following addition of purified MC LOS to epithelial cells. Using an MC mutant that completely lacks LOS (MC lps tbp), we examined pili-mediated attachment and cortical plaque formation in human endocervical epithelial cells (A431). We also tested whether the lack of LOS alters cellular homeostasis, including changes in the levels of host stress response factors and proinflammatory cytokines. MC lps tbp elicited the formation of cortical plaques in A431 cells. However, the plaques were less pronounced than those formed by the MC parent. Surprisingly, the proinflammatory cytokine TNF(alpha) was upregulated during infection in MC lps tbp-infected cells. Furthermore, alterations in iron homeostasis, including lower transferrin receptor 1 (TfR-1) levels, altered TfR-1 trafficking, an 'iron-starvation' gene expression profile and low iron regulatory protein (IRP) binding activity are independent of LOS. Our results demonstrate that LOS is partially involved in both the attachment to host cells and formation of cortical plaques. However, TNFalpha induction and changes in iron homeostasis observed in MC-infected epithelial cells are independent of LOS. PMID- 15888089 TI - Trichomonas vaginalis adherence mediates differential gene expression in human vaginal epithelial cells. AB - Trichomonas vaginalis, an ancient protist, colonizes the vaginal mucosa causing trichomonosis, a vaginitis that sometimes leads to severe health complications. Preparatory to colonization of the vagina is the adhesion to vaginal epithelial cells (VECs) by trichomonads. We hypothesized that VECs alter the gene expression to form a complex signalling cascade in response to trichomonal adherence. In order to identify the genes that are upregulated, we constructed a subtraction cDNA library after contact with parasites that is enriched for differentially expressed genes from the immortalized MS-74 VECs. Sixty cDNA clones were sequenced and to our knowledge for the first time, differentially regulated genes were identified in response to early trichomonal infection. The identified genes were found to encode functional proteins with specific functions associated with cell structure maintenance and extracellular matrix components, proinflammatory molecules and apoptosis. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) confirmed expression of selected genes. Further, cyclooxygenase 2 (COX-2) protein expression was analysed using Western blot and immunofluorescence assays. Data suggest that p38 mitogen-activated protein (MAP) kinase and tyrosine kinases play a role in COX-2 induction. Finally, T. vaginalis and Tritrichomonas foetus but not Pentatrichomonas hominis induce expression of COX-2. This is a first attempt at elucidating the basis of interaction of trichomonads with host cells and the corresponding host responses triggered by the parasites. PMID- 15888091 TI - Evaluation of simple non-invasive techniques for assessment of lower extremity arterial disease. AB - OBJECTIVE: To evaluate the reproducibility and precision of three, simple, non invasive methods to measure blood pressure (BP) in the lower extremities by comparing reproducibility and sensitivity in finding abnormally low BP between ankle blood pressure (ABP) and toe blood pressure (TBP), by studying the concordance between TBP in toe 1 and 2 and evaluating the pole-pox method in patients with diabetes and lower extremity arterial disease (LEAD). SUBJECTS AND METHODS: The BP was measured twice, 1 week apart, in arms and legs in 13 controls and 12 patients with diabetes. ABP was assessed by using a Doppler pen for pulse registration. TBP was obtained by using a small cuff and a pulse oximetry sensor at toe 1 and 2. In eleven patients with diabetes and previously known LEAD ABP was obtained through the pole-pox method. RESULTS: No significant difference in reproducibility between absolute BPs and indices (coefficients of variation <9%) was found. A non-significant improvement with 4-8% in the sensitivity in detecting LEAD was seen when BP indices were used instead of absolute BP. A significant correlation in the variation over time for systemic and TBP (r = 0.34, P = 0.015) and a strong correlation was found between TBP measured at toe 1 and 2, respectively (r = 0.99, P < 0.001) was found. TBP measured with pole-pox method were significantly correlated with measurements made by the ordinary cuff technique (r = 0.75, P < 0.001). CONCLUSIONS: The use of TBP and ABP indices instead of absolute BP does not improve the reproducibility but may improve the sensitivity with respect to detection of LEAD, especially in patients with diabetes. The pole-pox method may be used as an alternative screening method in patients with diabetes and LEAD. PMID- 15888092 TI - Evaluation of left ventricular volumes and ejection fraction by automated gated myocardial SPECT versus cardiovascular magnetic resonance. AB - BACKGROUND: Electrocardiogram-gated myocardial single-photon emission computed tomography (SPECT) with (99m)Tc-tetrofosmin allows simultaneous evaluation of myocardial perfusion and function. In this study, left ventricular volumes, ejection fraction (LVEF), and left ventricular wall volume (LVWV) derived from gated SPECT were compared with measurements from cardiovascular magnetic resonance (CMR), performed within a few hours. METHODS: The study population included 55 patients with known or suspected coronary artery disease, including 13 patients with recent acute myocardial infarction. End-diastolic (EDV) and end systolic (ESV) volumes, LVEF and LVWV were derived automatically from gated SPECT using commercially available software (QGS). In the CMR studies, manually delineated endocardial and epicardial borders on short-axis slices were used to calculate the volumes. RESULTS: Gated SPECT underestimated EDV by 35 +/- 14 ml (mean +/- SD) (P < 0.001), ESV by 10 +/- 13 ml (P < 0.001), and LVEF by 4 +/- 7 percentage points (P < 0.001). There were no systematic difference in EDV, ESV or LVEF between the methods. SPECT underestimated LVWV by 49 +/- 30 ml (P < 0.001), with a trend towards increasing underestimation by SPECT for larger wall volumes. CONCLUSION: These findings show that gated SPECT slightly underestimates EDV, ESV and LVEF compared with CMR. This underestimation is systematic, however, indicating that ventricular volumes derived from gated SPECT are robust enough to guide clinical management. Estimates of LVWV in patients with large wall volumes are less accurate. PMID- 15888093 TI - Effects of ventilation on cardiac output determined by inert gas rebreathing. AB - One of the most important methodological problems of the foreign gas rebreathing technique is that outcome of the measurements depends on procedural variables such as rebreathing frequency (RF), rebreathing bag volume (V(reb)), lung volume at start of rebreathing and intervals between measurements. Therefore, in 10 healthy males we investigated the effects of changes in ventilation pattern on cardiac output (CO) estimated by an N(2)O-rebreathing technique. Reducing the rebreathing volume (V(reb)) from 1.5 to 1.0 l diminished CO by 0.5 +/- 0.2 l min( 1), whereas an increase in V(reb) from 1.5 to 2.5 l had no effects. CO was 1.0 +/ 0.2 l min(-1) higher when, rebreathing was performed after a forced expiration than following a normal tidal expiration. Serial determinations of CO required a 3-min interval between the measurements to avoid effects of recirculation of N(2)O. Changing RF from 15 to 30 breaths min(-1) or adding serial dead space by up to 600 ml did not affect the determination of CO. In conclusion, the rebreathing procedure for determination of CO at rest should be performed following a normal tidal expiration with a rebreathing bag volume of between 1.5 and 2.5 l and with manoeuvres separated by at least 3-5 min. Variations in RF within the physiological range from 15 to 30 breaths min(-1) do not affect outcome of the measurements. PMID- 15888094 TI - A short period of apnoea causes a marked increase in coronary flow velocity: a transthoracic pulsed wave Doppler study. AB - Coronary artery flow velocity during a short period of apnoea was investigated by transthoracic Doppler recording in 10 healthy men, aged 24-52. During breath holding for 29 +/- 6 s the oxygen saturation in the finger, measured by pulse oxymetry, decreased from 97.5 +/- 0.8 to 90.0 +/- 3.2% (P < 0.001). The maximal coronary blood flow velocity in the left anterior descending artery increased by 62% from 0.26 +/- 0.09 to 0.42 +/- 0.10 m s(-1) and the mean diastolic flow velocity by 47% from 0.19 +/- 0.04 to 0.28 +/- 0.08 m s(-1) (P < 0.001). In most subjects the increase of velocity started already after a few seconds of apnoea. Besides theoretical implications the results show that it is very important to be aware of the higher velocity during apnoea in order to avoid misinterpretation when using breath holding during magnetic resonance imaging or Doppler recording of coronary flow. PMID- 15888095 TI - Effects of lung volume reduction surgery on distribution of ventilation and perfusion. AB - Lung volume reduction surgery (LVRS) results in functional improvements in most patients. The mechanisms behind the improvements are not clear. We hypothesized that reduced inequalities in ventilation to perfusion ratio (V/Q) may be a contributing explanation. Nine patients who underwent LVRS were investigated by ventilation and perfusion scintigrams before and after surgery. In addition, 8 healthy subjects were investigated once. The relative ventilation, perfusion and V/Q were calculated in 1 x 1 cm lung elements. Normal range of the element count rate was determined by the corresponding results in the normal subjects. Results of this small study show a significant effect of LVRS on V/Q, with reduction of shunt-like elements. We conclude that the functional improvement after LVRS to some extent may be explained by decreased V/Q inequality. PMID- 15888096 TI - Regional blood flow during hyperoxic haemodilution. AB - BACKGROUND: Ventilation with pure oxygen (hyperoxic ventilation, HV) increases arterial oxygen content (CaO(2)). However HV induces arteriolar constriction and thus potentially affects O(2) supply. We therefore investigated the effects of HV on regional blood flow (RBF) and O(2) supply of different vital organs during moderate normovolaemic anaemia. METHODS: Twenty-two anaesthetized dogs were haemodiluted under normoxia (i.e. FiO(2) = 0.21) to a target haemoglobin concentration (Hb) of 7 g dl(-1) and were subsequently ventilated with pure O(2). RBF was determined by use of the radioactive microspheres method in the myocardium, kidney, skeletal muscle, liver, intestine, stomach, and pancreas at Hb = 7 g dl(-1) and after subsequent initiation of HV. RBF in proportion to cardiac output (RBF(relative)), the variation coefficient of RBF (VC) and regional O(2) supply (rDO(2)) were calculated. RESULTS: Initiation of HV at Hb = 7.0 +/- 0.3 g dl(-1) reduced cardiac index (-17%) as well as RBF within the myocardium (-21%), pancreas (-25%), and skeletal muscle (-25%), whereas renal, hepatic, and intestinal RBF remained unchanged. Consequently RBF(relative) of the latter organs increased. Heterogeneity of RBF was marginally affected by HV. CONCLUSION: The initiation of HV during moderate normovolaemic anaemia (Hb =7 g dl(-1)) was accompanied by RBF redistribution with preference for renal, hepatic and intestinal O(2) supply. Cardiac, pancreatic and muscular O(2) supply decreased, however without any critical restriction of organ function. PMID- 15888097 TI - Parathyroid scintigraphy during hypocalcaemia in primary hyperparathyroidism. AB - BACKGROUND: Minimally invasive parathyroid surgery in patients with primary hyperparathyroidism (PHPT) demands high imaging accuracy. By increasing blood flow to the parathyroid adenoma before injection of a perfusion marker, we intended to improve the parathyroid scintigraphy. We have named the technique stimulated parathyroid scintigraphy (SPS). METHODS: Twenty minutes after injection of 100 MBq (99m)Tc-pertechnetate a thyroid scintigram was performed in 25 patients with PHPT. During the thyroid scintigraphy sodium citrate was infused which lowered plasma calcium by a mean of 14 +/- 1.3%. Then 700 MBq (99m)Tc sestamibi was injected and another scintigram of the neck was obtained. Perchlorate was given at the end of the sestamibi scintigram to increase the wash out of (99m)Tc-pertechnetate from the thyroid gland, and after 2 h a delayed scintigram was obtained. A subtraction of the thyroid scintigram from the initial sestamibi scintigram was performed. The results of SPS and a conventional (99m)Tc sestamibi dual-phase parathyroid scintigraphy were compared with the operative findings. In nine patients the parathyroid adenoma was also localized with ultrasound and the flow pattern before and after citrate infusion was visualized with Doppler technique. RESULTS: Eighty-eight per cent of the adenomas were localized correctly with the SPS technique compared with 62% at the conventional parathyroid scintigraphy. Tissue perfusion of the nine adenomas increased after citrate infusion. CONCLUSIONS: SPS has a high accuracy and it is easy to perform. If only subtraction SPS is performed the whole examination can be completed within an hour, which is acceptable for same day surgery. PMID- 15888098 TI - Left ventricular involvement in arrhythmogenic right ventricular cardiomyopathy - a scintigraphic and echocardiographic study. AB - BACKGROUND: Left ventricular involvement in arrhythmogenic right ventricular cardiomyopathy (ARVC) is a common finding in autopsy studies. In clinical studies using myocardial scintigraphy, MRI and echocardiography, contradictory results have been reported. In this study, we therefore investigated a group of 15 patients with ARVC using thallium-201 (Tl) single-photon emission tomography (SPECT) and echocardiography including assessment of mitral annular motion with M mode and pulsed tissue Doppler. METHODS: Exercise and rest Tl-201 SPECT were performed in 15 patients with ARVC. The time from diagnosis of the disease varied from less than 1-16 years. All patients fulfilled the established diagnostic criteria for ARVC. An echocardiographic examination, including assessment of left and right ventricular motion and measurements of the mitral annulus motion with M mode and pulsed tissue Doppler was performed in the patients and in 25 normal subjects. RESULTS: Tl-201 uptake defects in the left ventricular myocardium were present in all except one patient (93%). The uptake defects were predominantly located to the anteroseptal and basal posterior segments. Wall motion abnormalities were seen in the same segments, and in addition to this, in the septal area. In line with this, the total amplitude and the peak systolic velocity of mitral annular motion at the septal point were significantly decreased in the patients compared with the control group. CONCLUSIONS: Our data show that left ventricular involvement is common in ARVC. Tl-201 SPECT and echocardiographic abnormalities were seen not only in patients with long-lasting symptoms but also in asymptomatic patients and in those with short duration of symptoms. PMID- 15888099 TI - Comparison between maximal early diastolic velocity in long-axis direction obtained by M-mode echocardiography and by tissue Doppler in the assessment of right ventricular diastolic function. AB - Recently the maximal early diastolic velocity in long-axis direction of the right ventricle (RV) obtained by tissue Doppler imaging (MDV TDI) has been introduced in the assessment of RV diastolic function (RVDF). There are reasons to think that also the maximal early diastolic velocity in long-axis direction of the RV obtained using M-mode echocardiography (MDV TAM) could be used to assess RVDF. Therefore, 29 patients were examined with echocardiography and MDV TAM and MDV TDI were measured and compared. A good correlation (r = 0.76, P < 0.001) was found between MDV TAM and MDV TDI indicating that MDV TAM might be used in the assessment of RVDF. However, the velocities obtained by MDV TDI (126.7 +/- 38.9 mm s(-1)) were significantly (P < 0.001) higher than the velocities obtained by MDV TAM (78.3 +/- 27.8 mm s(-1)) and the agreement between MDV TAM and MDV TDI was rather poor probably mainly due to differences in the measuring technique. This means that reference values cannot be used interchangeably between MDV TAM and MDV TDI. If MDV TAM is going to be used in the assessment of RVDF new reference values have to be produced if today's technique and recommendations to measure MDV TAM and MDV TDI are used. However, as most new echocardiographs are equipped with PW-TDI technology it seems preferable to use this technique and compare obtained values with already established reference values. PMID- 15888100 TI - Detection of chronic rejection by quantitative ventilation scintigrams in lung transplanted patients: a pilot study. AB - The suspicion of chronic rejection [bronchiolithis obliterans syndrome (BOS)] is usually based on deteriorating forced expired volume in 1 s. It is however, desirable to develop more sensitive methods as increased anti-inflammatory therapy is thought to stop progression of the rejection. The aim of the present study was to develop quantitative tools based on ventilation scintigrams, to diagnose BOS. Sixteen double-lung-transplanted patients participated, six developing BOS and 10 who did not develop BOS. They were investigated with planar posterior-anterior (99m)Tc-Technegas (Tetley Manufacturing Ltd, Sydney, Australia) ventilation scintigraphy at baseline, 6 months to 1 year post transplantation, and at a follow-up examination 3-4-year post-transplant or in the BOS patients close to the time of the diagnosis. An automatic region of interest (ROI) was drawn on each lung in the scintigraphic image at baseline and also applied to the follow-up investigation. The area inside the ROI was subdivided into stripes 10.8 mm high and squares 10.8 x 10.8 mm wide. Corresponding stripes and squares in baseline and follow-up were analysed regarding differences in relative retention. The results show that the square analysis is superior. Applying chosen cut-off values for square element differences, 6/6 right and 5/6 left BOS lungs were identified and one left and one right lung of patients not developing BOS were misclassified. We conclude that the square element difference appears to be a promising method to diagnose BOS. PMID- 15888101 TI - Number II. Pemphigus vulgaris. AB - Pemphigus is a group of potentially life-threatening autoimmune diseases characterized by cutaneous and/or mucosal blistering. Pemphigus vulgaris (PV), the most common variant, is characterized by circulating IgG antibodies directed against desmoglein 3 (Dsg3), with about half the patients also having Dsg1 autoantibodies. There is a fairly strong genetic background to pemphigus with linkage to HLA class II alleles and ethnic groups such as Ashkenazi Jews and those of Mediterranean and Indian origin, are especially liable. Oral lesions are initially vesiculobullous but readily rupture, new bullae developing as the older ones rupture and ulcerate. Biopsy of perilesional tissue, with histological and immunostaining examination are essential to the diagnosis. Serum autoantibodies to either Dsg1 or Dsg3 are best detected using both normal human skin and monkey oesophagus or by enzyme-linked immunosorbent assay. Before the introduction of corticosteroids, PV was typically fatal mainly from dehydration or secondary systemic infections. Current treatment is largely based on systemic immunosuppression using corticosteroids, with azathioprine or other adjuvants or alternatives but newer therapies with potentially fewer adverse effects, also appear promising. PMID- 15888102 TI - Bacteriotherapy and probiotics' role on oral health. AB - Oral infections constitute some of the most common and costly forms of infections in humans. The concept of microbial ecological change as a mechanism for preventing dental disease is an important one while altered microbial ecology may lead to dental disease. New methods such as probiotic approaches (i.e. whole bacteria replacement therapy) to eliminate pathogenic members of the microbiota can be investigated. Bacteriotherapy is an alternative and promising way to combat infections by using harmless bacteria to displace pathogenic microorganisms. Probiotics are one of these new agents which are widely used for their therapeutic action. Limited research is available showing that some probiotic cultures may help dental improvement. Present paper focuses on possible oral benefits of probiotics. PMID- 15888103 TI - Oral sarcoidosis: a review of literature. AB - Sarcoidosis is a common systemic granulomatous disease affecting multiple organs. Oral involvement is relatively rare and, to our knowledge, there have been only 64 cases reported in the English literature. Most cases of oral sarcoidosis present with mobility of the teeth due to rapid alveolar bone loss. Other oral manifestations include asymptomatic swelling of the involved mucosa, gingivitis and ulcers. Diagnosis of sarcoidosis is by exclusion as no specific test is available. Radiographic, biochemical and histological findings are non-specific, but helpful. All cases of sarcoidosis do not require treatment. Corticosteroids are the treatment of choice in patients requiring treatment. Other drugs such as chloroquine, methotrexate, infliximab and thalidomide are also used in the treatment of sarcoidosis. In most of the oral cases reported, systemic steroids and surgery were the preferred treatment. PMID- 15888104 TI - Chlorhexidine digluconate induces mitotic recombination in diploid cells of Aspergillus nidulans. AB - Chlorhexidine digluconate (1,1'-hexamethylene-bis[(5-p-clorophenyl)-biguanide]) is a bisbiguanidine antiseptic, used to decrease plaque formation and to control periodontal diseases. The determination of the frequency of mitotic crossing-over constitutes a very important method for detecting carcinogenic agents. OBJECTIVE: The recombinogenic potential of chlorhexidine digluconate was evaluated on Aspergillus nidulans by the production of cells homozygous for the following nutritional markers: riboA1, pabaA124, biA1, methA17 and pyroA4. METHOD: A. nidulans was exposed to three concentrations of chlorhexidine digluconate (1, 5, and 10 microM). RESULTS: Inhibition of colony development, conidiophore morphological alteration (cytotoxic effect), and the recombinogenic effect, indicated by homozygotization index (HI) values higher than 2.0, were observed for all concentrations of chlorhexidine digluconate. A homozygous pyro+//pyro+ diploid strain and a diploid homozygous for the recessive w gene were isolated from UT448//A757 diploid treated with chlorhexidine digluconate, emphasazing its recombinogenic potential. CONCLUSION: Although, beneficial effects of chlorhexidine, as an antiseptic agent, are reported in the literature, our results revealed that chlorhexidine digluconate, at less levels lowered those used clinically, caused toxic and recombinogenic effects on diploid A. nidulans strain. PMID- 15888105 TI - Streptococcus anginosus infection in oral cancer and its infection route. AB - OBJECTIVE: To elucidate a possible involvement of Streptococcus anginosus in oral cancer, we assessed the frequency of S. anginosus infection in oral cancer tissues, and investigated its infection route. MATERIALS AND METHOD: The tissue specimens were obtained from 46 oral cancer and three precancerous leukoplakia subjects. Frequency of S. anginosus infection was assessed by a species-specific polymerase chain reaction (PCR) assay. The genotype of the clinical isolates taken from cancer tissue and dental plaque samples was analyzed using pulsed field gel electrophoresis (PFGE). RESULTS: S. anginosus DNA was frequently detected in squamous cell carcinoma (19/42), but not in other types of cancer (lymphoma and rhabdomyosarcoma) or leukoplakia samples. A subject-based analysis revealed that S. anginosus was solely detected in dental plaque and not in saliva from all 19 S. anginosus-positive squamous cell carcinoma cases. Further, the genotype of S. anginosus isolated from cancer tissue was identical to that from dental plaque of the same patients. CONCLUSION: Infection of S. anginosus could occur frequently in oral squamous cell carcinoma and that dental plaque could be a dominant reservoir of the S. anginosus. PMID- 15888106 TI - Prostaglandin E2 (PGE2) downregulates interleukin (IL)-1alpha-induced IL-6 production via EP2/EP4 subtypes of PGE2 receptors in human periodontal ligament cells. AB - OBJECTIVES: Prostaglandin E2 (PGE2) exerts its biological actions via EP receptors, which are divided into four subtypes, EP1, EP2, EP3 and EP4. In the present study, we examined whether PGE2 regulated interleukin (IL)-1alpha-induced IL-6 production in human periodontal ligament (PDL) cells and if so, which subtypes of PGE2 receptors were involved. METHODS: PDL cells were stimulated with vehicle or IL-1alpha in the presence or absence of indomethacin (a cylooxygenase inhibitor), PGE2 or various EP agonists. IL-6 and PGE2 levels were measured by enzyme-linked immunosorbent assay. EP receptor mRNA expression was examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Indomethacin significantly enhanced IL-1alpha-induced IL-6 production by PDL cells, although it completely inhibited IL-1alpha-induced PGE2 production. Exogenous PGE2 significantly suppressed IL-1alpha-induced IL-6 production. Butaprost, a selective EP2 agonist, and ONO-AE1-329, a selective EP4 agonist, significantly inhibited IL-1alpha-induced IL-6 production, although 17-phenyl-omega-trinor PGE2, an EP1 agonist, and ONO-AP-324, an EP3 agonist, did not affect it. RT-PCR analysis showed that EP2 and EP4 mRNA was expressed in PDL cells. CONCLUSIONS: We suggest that PGE2 downregulates IL-1alpha-induced IL-6 production via EP2/EP4 receptors in human PDL cells. PMID- 15888107 TI - Improved oral hygiene and Candida species colonization level in geriatric patients. AB - OBJECTIVES: This work consists in improving oral hygiene (OH) for elderly dependent people in long-term hospital care, in order to decrease the degree of colonization and the associated risk of developing oral candidiasis. As this population frequently suffers from such colonization and because it is difficult to install and practice OH care, a study protocol was designed at the request of geriatricians. The objective of the present study was to set up a programme of OH, applied by the care staff, and to monitor oral colonization of by Candida spp. BASIC RESEARCH DESIGN: We compared the levels of hygiene and Candida spp. colonization for a group of 110 long-term patients in geriatric departments at T1, when clinical data were collected and oral mycological samples taken before the OH protocol was applied, and at T2, during the postprotocol phase after 3 months of application, when the clinical data and sample collection were repeated. RESULTS: During these 3 months 11 patients died. These patients were excluded from the results, which are presented for matched series of the 99 patients still present at T2. Statistical analysis comparing the clinical and biological parameters at T1 and T2 established that there had been an improvement in OH: the 'adequate' level was reached for 72.4% of patients at T2 compared with 41.8% at T1 (P < 0.001) and the 'very inadequate' level was observed for 9.2% at T2 compared with 27.9% at T1 (P < 0.01). A reduction was observed in the number of patients showing the highest degree of C. albicans and C. glabrata colonization (> 50 colony forming units) from 41.9% at T1 to 24.9% at T2 (P < 0.05) and from 56.4% at T1 to 13.0% at T2 (P < 0.05) respectively. The number of patients with candidiasis fell significantly from 43.2% at T1 to 10.2% at T2. CONCLUSIONS: The OH protocol led to an overall decrease in Candida spp. colonization, a significant reduction in the number of candidiasis and an improvement in the level of oral and denture hygiene but vigilance is still necessary concerning OH care and the initial training of staff in specific care of the mouth. PMID- 15888108 TI - Ankyloglossia: a morphofunctional investigation in children. AB - OBJECTIVES: To provide diagnostic criteria for ankyloglossia in children by anatomical measurements; to investigate the correlation between severity of ankyloglossia and a series of morphofunctional findings; to evaluate the potential mismatch between a clinical suspect of ankyloglossia and the authentic anatomical diagnosis. DESIGN: Two different techniques of anatomical measurements and a clinical evaluation of a series of morphofunctional findings were performed. SUBJECTS AND METHODS: In 200 children referred for evaluation and treatment of tongue-tie, the length of the frenulum and the interincisal distance were measured in maximum opening of the mouth and with the tip of the tongue touching the palatal papilla. Occlusion, type of bite, tongue resting position, swallowing mechanism, oral floor mobility, frenulum insertion modality and speech were investigated. Any correlation between these morphofunctional findings and anatomical measures was investigated. RESULTS: Children with a frenulum length more than 2 cm and an interincisal distance of more than 2.3 cm were normal. In both measurements, significant correlations among mean values and other variables were observed. Moreover, three levels -- mild, moderate and severe -- of ankyloglossia were assessed. CONCLUSIONS: Length of frenulum and interincisal distance allow an assessment of severity of ankyloglossia in children. Ankyloglossia was not associated with infantile swallowing. PMID- 15888109 TI - Adenoid cystic carcinoma and polymorphous low-grade adenocarcinoma of minor salivary glands: a comparative immunohistochemical study using the epithelial membrane and carcinoembryonic antibodies. AB - OBJECTIVE: The purpose of this study was to investigate immunohistochemically the expression of epithelial membrane antigen (EMA) and carcinoembryonic antigen (CEA) in adenoid cystic carcinoma (AdCC) and polymorphous low-grade adenocarcinoma (PLGA) in an attempt to assess the ability of these markers to distinguish AdCC from PLGA when the histological features on routine hematoxylin and eosin are equivocal. MATERIALS AND METHODS: Fourteen specimens of AdCC, 10 PLGA, and five normal minor salivary glands fixed in 10% formalin and embedded in paraffin, were retrieved from the files of our department and were retrospectively studied with the streptavidin-biotin complex method using the epithelial membrane and carcinoembryonic antibodies. RESULTS: The immunoreactivities and the expression patterns of EMA and CEA in AdCC and PLGA were similar. CONCLUSIONS: The results of this study suggest that the immunostaining of AdCC and PLGA with EMA and CEA could not offer an adjunctive aid in differential diagnosis between these two tumors. PMID- 15888110 TI - HPV and other risk factors of oral cavity/oropharyngeal cancer in the Czech Republic. AB - OBJECTIVE: An association between high-risk human papillomavirus (HR HPV) infection and a risk of development of a subgroup of head and neck cancers has been proposed recently. The main risk factors of oral and oropharyngal cancer observed in our population are smoking and alcohol consumption. The incidence of oral/oropharyngeal tumours in the Czech Republic is relatively high and there are no data available about the prevalence of HPV DNA presence in these tumours. MATERIALS AND METHODS: Eighty patients with a primary oropharyngeal cancer were enrolled. The presence of HPV DNA has been evaluated by polymerase chain reaction in 68 cases from which the tumour tissue and demographical and clinical data were available. The typing of HPV was performed by nucleotide DNA sequencing. RESULTS: The HPV DNA was detected in 51.5% of samples tested. Among the HPV DNA positive tumours, 80% contained HPV16. In the analysed group there were 54 men and 14 women. The prevalence of HPV DNA was lower in oral (25%) than in oropharyngeal (57%) tumours, and higher in never smokers (100%) and never drinkers (68.8%). HPV DNA presence was not related to gender, age, number of lifetime sexual partners or practice of oral-genital sex, size of tumour or presence of regional metastases. CONCLUSIONS: The difference in the prevalence of HPV DNA positive tumours between cases of oral cavity and oropharyngeal carcinoma exposed and not exposed to tobacco or alcohol support the theory that HPV DNA positive tumours form an aetiologically distinct subgroup of head and neck tumours. PMID- 15888111 TI - Trismus-pseudocamptodactyly syndrome (Hecht-Beals' syndrome): case report and literature review. AB - The trismus-pseudocamptodactyly syndrome (TPS) is a rare condition inherited as an autosomal dominant trait with variable expressivity. Clinically the syndrome is characterized by decreased ability to open the mouth and curvature of the fingers at the level of interphalangic joints while attempting dorsiflexion of the wrist (pseudocamptodactyly). A 6-year-old male patient from Guatemala with this syndrome is presented. The surgical treatment consisted of bilateral coronoid amputation to resolve the pseudotrismus. The procedure was successful and the patient achieved normal function. PMID- 15888112 TI - Congenital heart block associated with undiagnosed maternal Primary Sjogren's Syndrome -- a case report and discussion. AB - Congenital heart block (CHB) has been linked with Sjogren's Syndrome. This paper reports a case of previously undiagnosed maternal Primary Sjogren's Syndrome (1 degrees SS) that was only discovered following the birth of the patient's first child with CHB. The possible pathophysiological mechanisms underlying CHB associated with 1 degrees SS are discussed. PMID- 15888114 TI - In reply: Pleomorphic adenoma with extensive necrosis report of two cases. Oral Diseases 2004; 10: 54-59. PMID- 15888116 TI - How best to 'time' education. PMID- 15888117 TI - Stem cells in craniofacial and dental tissue engineering. AB - Mesenchymal stem cells (MSC) have been identified in a variety of adult tissues as a population of pluripotential self-renewing cells. Based on their adherence and colony forming properties, a small number of MSC can be isolated from most mesenchymal tissues as well as bone marrow. In the presence of one or more growth factors, these cells commit to lineages that lead to the formation of bone, cartilage, muscle, tendon and adipose tissue; recent studies indicate that stem cells for cementum, dentine and the periodontal ligament also exist. All of these cells can be expanded in vitro, and, embedded in a scaffold, inserted into defects to promote healing and tissue replacement. Increased understanding of the molecular mechanism directing lineage specification and morphogenesis is providing a rational approach for the regeneration of craniofacial tissues and oral structures. PMID- 15888118 TI - Herbal therapy: a review of potential health risks and medicinal interactions. PMID- 15888119 TI - Tolerance of deviations in eye and mouth position. AB - OBJECTIVE: To develop and refine a technique for measuring the tolerance for deviations in facial appearance. DESIGN: A psychophysical method was administered using photocopies of altered facial photographs to measure tolerances for deviations in eye position and mouth angulation and judges' reaction time. METHODS: Stimulus photos were displayed as Kodachrome slides to 76 individuals grouped by their familiarity with craniofacial anomalies, i.e. dental professionals, orthodontic and craniofacial patients, also a group of normal patients. RESULTS: (1) Tolerance for deviation of facial appearance varies inversely with the magnitude of the physical deviations from normal; (2) tolerance varies directly as a function of assumed familiarity with deviation; (3) response and reaction time varied inversely with the tolerance for facial deviation. CONCLUSION: Significant differences in tolerance and reaction time were found among the groups and depended on whether an isolated feature was judged or if the feature was judged in the context of the whole face. PMID- 15888120 TI - The investigation of the changing facial appearance of identical twins employing a three-dimensional laser imaging system. AB - OBJECTIVE: An investigation to determine the changing facial appearance of identical twins. DESIGN: Clinical study. MATERIALS AND METHODS: Two Minolta Vivid 900 3D optical laser scanners were placed as a stereo pair to capture the soft tissues of a pair of identical twins. Each scan took approximately 2.5 s. The scanned whole faces were superimposed to determine changes in facial morphologies at different time intervals. OUTCOME MEASURES: The shell deviations between left and right scans of each patient were recorded and analysed for differences. Furthermore, final merged faces were overlaid to determine the changes in facial morphology over time. RESULTS: The results showed that changes in height and weight correlated with changes in facial morphology. CONCLUSION: The 3D laser scanning device is a clinically useful tool in the study of facial growth and facial morphology in a pair of twins. PMID- 15888121 TI - Short hard palate in prenatal trisomy 21. AB - OBJECTIVE: The aim of the present study was for the first time to examine on postmortal material the total midpalatal length of the hard palate and the length of its two components (the maxillary and palatine parts) in trisomy 21 fetuses, and to compare the results to normal standards. DESIGN: Material from 31 human fetuses with genetically verified trisomy 21 was studied. The fetuses were derived from legally induced or spontaneous abortions. Palates were, after sectioning, radiographed in lateral projection (Grenz Ray radiographic apparatus). Cephalometric measurements were performed with a digital caliper. Statistically, the length measurements for the two groups were compared, adjusting for crown rump length (CRL) through linear regression. At two specific ages (150 and 170 mm CRL), the length of the palatal components in trisomy 21 was compared to normal standards. RESULTS: For CRL 150 mm and CRL 170 mm it appears that all three palatal lengths, total length, maxillary length, and palatinal length are significantly shorter in fetuses with trisomy 21. CONCLUSION: The main conclusion of our study is that the total palatal length in prenatal trisomy 21 is shorter than normal and that this is due both to a shortness of the maxillary and the palatine components of the hard palate. PMID- 15888122 TI - In vitro friction of stainless steel arch wire-bracket combinations in air and different aqueous solutions. AB - OBJECTIVES: To investigate the in vitro coefficient of friction of stainless steel arch wire-bracket combinations under fretting contact test conditions performed in air and in different aqueous solutions, like Ringer solution, Ringer with addition of a buffer, Ringer with addition of glucose, and Coca Cola. METHODS: The fretting test set-up used allowed to control on-line the contact configuration and the positioning of the contacting parts. A specific positioning method was used to achieve a parallel alignment of arch wire and bracket slot. The effect of arch wire size, roughness, and test environment were investigated. RESULTS: It was found that the aqueous solutions act as a lubricant compared to air. Friction was affected by the arch wire width while the roughness was found to have a limited effect. Stainless steel 0.018'' x 0.025'' arch wires exhibited higher frictional forces than stainless steel 0.017'' x 0.025'' arch wires on sliding against stainless steel 0.018'' x 0.025'' brackets in the selected test environments when tested under identical fretting test conditions. The wear damage on the arch wire after these in-vitro fretting tests was investigated. It revealed that these in-vitro tests are governed by a competition between oxidational wear and abrasive wear taking place at contact areas between brackets and arch wires. CONCLUSIONS: For all aqueous solutions a lower coefficient of friction was found compared to tests performed in ambient air. PMID- 15888123 TI - Temporary loss of plasma membrane integrity in orthodontic tooth movement. AB - In these studies, a rat model of orthodontic tooth movement was used to support the premise that periodontal ligament (PDL) cells experience plasma membrane disruption and resealing events upon application of mechanical stress. Immunoelectron microscopy, showed albumin in the cytoplasm of PDL and bone lining cells in the tension side of moved molars. The intracellular localization of this large molecule (60 KDa) suggests that these cells have undergone plasma membrane disruption and resealing. To further assess these and previous findings, fluorescent dyes (FITC-dextran and rhodamine-dextran) were delivered into the vascular system followed by application of 50 g of static load. These large dextran molecules (10 KDa) were preferentially taken up by PDL cells of the buccal (tension side) of moved molars. These cells were determined to be viable since dead cells do not retain these diffusible tracers. These studies provide evidence of a novel cellular mechanism for uptake and release of molecules and suggest a potential role for plasma membrane disruption in the mechanotransduction of orthodontic tooth movement. PMID- 15888124 TI - Identification of temporal pattern of mandibular condylar growth: a molecular and biochemical experiment. AB - OBJECTIVES: Based on the phenomenon that expression of type X collagen and capillary endothelium correlates with endochondral ossification, the prime aim of this study was to establish the temporal pattern of condylar growth in Sprague Dawley rats by biochemically identifying the expression of these two factors. DESIGN: Sprague-Dawley rats were divided into five groups representing five different stages during somatic pubertal growth. In situ hybridization and immunoperoxidase were performed to examine expression of type X collagen in hypertrophic zone and capillary endothelium in erosive zone of condylar cartilage. Computer-assisted imaging analyses were conducted to allow for a quantitative assessment of the expression of these two factors, from which the temporal pattern of condylar growth was inferred. RESULTS: (1) Synthesis of type X collagen and emergence of capillary endothelium were critical factors during the transition of condylar cartilage from chondrogenesis into osteogenesis, a biological pathway that leads to endochondral bone formation, the mode through which the condyle grows. (2) Quantitative analyses revealed the temporal pattern of the expression of these two factors, indicating that the thrust of natural growth of the condyle in the rats occurred in concomitance with somatic pubertal growth, featured by an acceleration starting from day 38, a maximum growth rate on day 56, followed by a decrease afterwards. CONCLUSION: It is suggested that the biochemical examination of growth markers, such as type X collagen, might be a new approach to accurately depict temporal pattern of condylar growth which is too delicate to be reflected by gross measurement not only in Sprague-Dawley rats but potentially also in other species. PMID- 15888125 TI - Interaction between Smad 3 and Dishevelled in murine embryonic craniofacial mesenchymal cells. AB - OBJECTIVES: To determine the in vivo interaction between Smad 3 and Dishevelled 1. DESIGN: Cell culture transfection followed by immunoprecipitation with specific antibodies. SETTING AND SAMPLE POPULATION: The Department of Molecular, Cellular, and Craniofacial Biology, Birth Defects Center, University of Louisville. EXPERIMENTAL VARIABLE: Overexpression of myc-Smad 3. OUTCOME MEASURE: Western blotting of anti-Dishevelled immunoprecipitates for Smad 3. RESULTS: Smad 3 and Dishevelled isoforms-1, -2, and -3 all bind Smad 3 in glutathione-S transferase (GST) pull-down assays and Smad 3 binds to Dishevelled-1 in vivo. Stimulation of the transforming growth factor beta (TGFbeta) pathway leads to increased binding of Smad 3 and Dishevelled-1 in vivo. CONCLUSION: Smad 3 binds all three known isoforms of Dishevelled and binds Dishevelled 1 in vivo. TGFbeta signaling modulates the interaction between Smad 3 and Dishevelled-1. PMID- 15888126 TI - A new light on Niels Finsen, a century after his Nobel Prize. PMID- 15888127 TI - How Finsen's light cured lupus vulgaris. AB - In 1903, Niels Ryberg Finsen was awarded the Nobel Prize for his invention of light therapy for skin tuberculosis (lupus vulgaris). The mechanism of action has not been shown; thus, we wanted to elucidate the mechanism of Finsen's light therapy. We measured radiation that could be transmitted through his lens systems and absorption of the stain solution filters in the lamps, and related the obtained results to the possible biological effects on Mycobacterium tuberculosis. Judged from transmission characteristics all tested lens systems were glass lenses (absorbing wavelength < 340 nm). The tested filters likewise absorbed wavelengths < 340 nm. The methylene blue solution used to absorb heat, blocked out wavelengths below 340 nm and between 550 and 700 nm. Furthermore, fluorescence of M. tuberculosis indicated the presence of porphyrins and HPLC analysis of sonicated M. marinum showed that coproporphyrin III was present, which highly justified that porphyrins were present in M. tuberculosis. Production of singlet oxygen through radiation of porphyrins with light of e.g. 400 nm seems to be a most plausible explanation why Finsen's therapy worked in spite of the lack of shortwave ultraviolet radiation, which Finsen believed was the most effective radiation for treating skin tuberculosis. PMID- 15888128 TI - Therapeutic and prophylactic effects of PUVA photochemotherapy on atopic dermatitis-like lesions in NC/Nga mice. AB - BACKGROUND: Psoralens and ultraviolet A radiation (PUVA) photochemotherapy has been used for severe cases of atopic dermatitis (AD). To understand the mechanisms of action is important for the choice of treatments. AD-like lesions can be induced experimentally in NC/Nga mice. OBJECTIVES: To evaluate clinically and histologically the therapeutic and prophylactic effects of PUVA on AD-like dermatitis using NC/Nga mice. METHODS: PUVA therapy was performed with intraperitoneal injection of 4 mg/kg of 8-methoxypsoralen (8-MOP) and 4 J/cm(2) UVA irradiation before and after development of AD-like lesions in NC/Nga mice which had been maintained in a conventional room (Conv-NC/Nga mice). Clinical skin conditions were evaluated periodically by a clinical severity score defined. Lesions were histologically examined in haematoxylin-eosin or toluidine blue stained sections. Plasma levels of total IgE were measured at various time points. RESULTS: In Conv-NC/Nga mice infested with mite, AD-like lesions started to develop at 8 week of age and thereafter increased in severity score. PUVA therapy at lower does than minimal phototoxic dose suppressed the development of dermatitis and was also therapeutically effective against established lesions. Proliferation of dermal mast cells in AD-like lesions was suppressed, but IgE hyperproduction was not changed after PUVA. CONCLUSIONS: These observations suggest that PUVA photochemotherapy reveals not only therapeutic but also prophylactic effects on human AD. PMID- 15888129 TI - A study of Q-switched Nd:YAG laser irradiation and paracrine function in human skin cells. AB - BACKGROUND AND OBJECTIVES: This preliminary laboratory-based study looks at the paracrine release from human skin cells subject to sublethal Q-switched Nd:YAG 532 nm laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: Human dermal fibroblast and keratinocyte cultures were exposed to sublethal energy using the Nd:YAG 532 nm laser. Altered gene expression was then screened using RT-PCR for a range of paracrine factors known to affect melanogenesis, basic fibroblast growth factor (b-FGF), hepatocyte growth factor (HGF), stem cell factor (SCF), melanocyte stimulating hormone (MSH), endothelin-1 (ET-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and protease-activated receptor-2 (PAR 2). Enzyme-linked immunosorbent assay (ELISA) was used to confirm protein production. Conditioned medium was used to assess altered melanogenesis in a melanoma cell line. RESULTS: Fibroblasts exposed to sublethal radiation showed upregulation of b-FGF, HGF and SCF. This contrasts with keratinocytes which showed upregulation of IL-6. Elevated protein levels of b-FGF and SCF were confirmed by ELISA assay. Conditioned fibroblast medium was shown to stimulate melanogenesis in a melanoma cell line. CONCLUSIONS: This preliminary laboratory study reports, for the first time, specific gene upregulation using the Q switched Nd:YAG 532 nm laser. PMID- 15888130 TI - Evaluation of vaseline oil applied prior to UVB TL01 phototherapy in the treatment of psoriasis. AB - BACKGROUND: Use of emollient prior to phototherapy could enhance UV transmission through psoriasis plaques on the condition that the emollient is not photoprotective. Emollient pretreatment with narrow-band phototherapy (313 nm) has not been studied extensively. We conducted a study to assess if vaseline oil prior to UVB TL01, in chronic psoriasis plaques could accelerate psoriasis clearance. METHODS: Fifteen patients with chronic psoriasis plaques were enrolled in a prospective, single-blind, controlled study. For each patient, one to three symmetrical pairs of plaque were selected and scored initially and after every six exposures. RESULTS: On the vaseline oil pretreated side, significantly more plaques were cleared, especially in severe psoriasis. Scaling and infiltration were significantly improved. Application of vaseline oil was more interesting in thick and scaly psoriasis probably because the oil penetrates the intercellular space allowing an optical matching effect which increases the UV transmission. CONCLUSION: We strongly recommend vaseline oil pretreatment with UVB TL01 phototherapy in psoriasis, especially in severe psoriasis. PMID- 15888131 TI - Photodynamic therapy in dermatology--an update. AB - Topical photodynamic therapy (PDT) is a well-established treatment modality which has mainly shown to be effective for dermatooncologic conditions like actinic keratoses (AK), Bowen's disease, in situ squamous cell carcinoma and superficial basal cell carcinoma (BCC). However, a therapeutical benefit of PDT is also evident for inflammatory dermatoses like localized scleroderma, acne vulgaris and granuloma annulare. Recent work has been focused on the development and evaluation of topical photosensitizers like the heme precursor 5-aminolevulinic acid (5-ALA) or its methyl ester (methyl aminolevulinate) inducing photosensitizing porphyrins. These drugs do not induce strong generalized cutaneous photosensitization like the systemically applied porphyrins or their derivatives. For dermatological purposes, incoherent lamps or light-emitting diode arrays can be used for light activation. Depending on the applied light dose and the concentration of the photosensitizer either cytotoxic effects resulting in tumor destruction or immunomodulatory effects improving the inflammatory conditions occur. Treating superficial oncologic lesions (tumor thickness <2-3 mm) cure rates achieved by PDT are equal to the cure rates of the respective standard therapeutic procedure. The benefits of PDT are the low level of invasiveness and the excellent cosmetic results after treatment. PMID- 15888132 TI - The course of anticoagulation after extracorporeal photochemotherapy. AB - During extracorporeal photochemotherapy (ECP) heparin is added to the extracorporeal circulation to avoid clotting. It has not been investigated whether and to what extend this procedure influences coagulation in patients after ECP. In this study activated partial thrombaplastin time (aPTT) and anti factor Xa were monitored for up to 4 h after ECP in 10 patients. We found that anticoagulation followed the typical course of intravenously applied heparin: immediately after reinfusion anti-Xa was high (>1 IU/ml) and aPTT was prolonged to more than 180 s followed by a rapid decline and reaching normal values within 3-4 h in all patients. We conclude that anticoagulation and increased risk of bleeding after ECP is confined to a short period immediately following treatment. Physicians using ECP should be aware of the course of anticoagulation associated with this treatment. PMID- 15888133 TI - Exacerbation of presumed chronic actinic dermatitis by cockpit visible light in an airline pilot with atopic eczema. AB - Chronic actinic dermatitis (CAD) is a persistent ultraviolet radiation- or visible light-induced eczema of predominantly the exposed areas of usually elderly people. We now present the case of a young pilot with atopic eczema who developed CAD, regularly exacerbated by exposure to visible light through his aircraft cockpit window. PMID- 15888134 TI - Refractory livedoid vasculitis responding to PUVA: a report of four cases. AB - Livedoid vasculitis is a chronic disease characterized by recurrent painful irregularly shaped ulcers, which heal with scars, most commonly located on feet or lower extremities. This condition is often resistant to the therapy. We report four cases with refractory livedoid vasculitis that responded to systemic psoralens and ultraviolet A radiation therapy. PMID- 15888135 TI - Phototherapy and photochemotherapy of sclerosing skin diseases. AB - The treatment of sclerosing skin diseases [systemic sclerosis, localized scleroderma, lichen sclerosus et atrophicus, sclerodermoid graft-vs.-host disease, scleredema adultorum (Buschke), scleromyxedema and necrobiosis lipoidica] is difficult and remains a great challenge. Numerous treatments, some with potentially hazardous side effects, are currently used with only limited success. The introduction of phototherapy and photochemotherapy for sclerosing skin diseases has considerably enriched the therapeutic panel and proven useful in a number of sclerosing skin diseases especially in localized scleroderma. Two phototherapeutic modalitites are used for the treatment of sclerosing skin diseases, long-wave ultraviolet A and psoralen plus ultraviolet A (PUVA). This article reviews current knowledge about the application of phototherapy and photochemotherapy to various sclerosing skin disorders. PMID- 15888137 TI - Compulsory registration of clinical trials. PMID- 15888138 TI - Current severe psoriasis and the rule of tens. AB - This review addresses the problems of defining severity of psoriasis. Concepts of severity depend on the timescale perspective from which judgement is made. Measurement needs to include assessment of signs, impact on the patient's life and the history of the disease. The concept of severity in relationship to quality of life measurement scores has been defined, so it is now possible to postulate a standard, easily remembered concept to help define 'severe psoriasis' in the clinic. The proposed Rule of Tens for current severe psoriasis from the clinician's viewpoint is: 'Current Severe Psoriasis = Body Surface Area involved > 10% or Psoriasis Area and Severity Index score > 10 or Dermatology Life Quality Index score > 10'. PMID- 15888139 TI - PTCH codon 1315 polymorphism and risk for nonmelanoma skin cancer. AB - BACKGROUND: The PTCH tumour suppressor gene is involved in the development of nearly all basal cell carcinomas (BCCs) of the skin and a fraction of squamous cell carcinomas (SCCs). A nonconservative Pro/Leu nucleotide polymorphism within PTCH exon 23 at codon 1315 was recently reported to be potentially important for the development of breast epithelial cell cancers. Objectives Accordingly, the status of PTCH codon 1315 was analysed for a possible association with the development of nonmelanoma skin cancers (NMSCs) in a pilot study. Because skin cancer risk is affected by specific population-dependent phenotypes such as skin and hair colour, codon 1315 was also analysed for normal allele frequency variation in human populations having differing extents of eumelanin vs. phaeomelanin. METHODS: The single nucleotide polymorphism in codon 1315 of the human PTCH gene was analysed in genomic DNA from six different populations comprising 472 blood samples and from 170 patients in four different categories with NMSC. Polymerase chain reaction and pyrosequencing were used to determine the allele frequencies. Allelic loss was furthermore determined in tumours following microdissection. RESULTS: The Pro/Pro genotype frequency ranged from 30% to 65% between populations, with a significant trend for a reduced frequency of the Pro/Pro genotype in populations having lighter pigmentation (P = 0.020). Pro/Pro frequency showed an increasing trend with increasing tumour case severity (P = 0.027). In 260 samples from 180 Swedish patients with NMSC and a control group of 96 healthy ethnically matched volunteers, no statistically significant pairwise differences between groups were detected in the PTCH codon 1315 allelic distribution, neither was a difference seen for multiple or early onset cases of BCC in the Swedish population. In Swedish patients with single tumours, allelic loss (loss of heterozygosity) was observed in 20 of 30 (67%) patients with BCC and four of 22 (18%) patients with SCC, with no preference in the allele lost. In contrast, the Pro/Pro genotype was frequent in seven U.S. patients having multiple independent BCCs. One of these patients was heterozygous, enabling allelic loss studies. Of 20 independent tumours, 11 had lost an allele; 10 of the 11 had lost Leu, suggesting nonrandom loss that favoured retention of Pro (P = 0.0059). CONCLUSIONS: Our results indicate an association between the eumelanin to-phaeomelanin shift and a shift from the Pro/Pro genotype to Leu-containing genotypes. Failure to lose Pro during the shift to phaeomelanin may be associated with an increased population risk for BCC and increased individual risk for multiple BCC. During development of a tumour, the effect of Pro may be magnified by loss of the Leu allele. PMID- 15888140 TI - Refined localization of a punctate palmoplantar keratoderma gene to a 5.06-cM region at 15q22.2-15q22.31. AB - BACKGROUND: Punctate palmoplantar keratoderma (PPK) is a rare autosomal dominant cutaneous disorder characterized by numerous hyperkeratotic papules distributed on the palms and soles. Two loci for punctate PPK were recently found to be located on 8q24.13-8q24.21 and 15q22-15q24. However, no genes for this disease have been identified to date. Objectives To refine the previously mapped regions and to identify the disease gene locus in a four-generation Chinese family with punctate PPK. METHODS: Genetic linkage analysis was carried out in this family using microsatellite markers on chromosomes 8q and 15q. Two-point linkage analysis was performed using Linkage programs version 5.10 and the haplotype was constructed using Cyrillic version 2.02 software. RESULTS: We failed to confirm our previous locus at 8q24.13-8q24.21, but significant evidence for linkage was observed in the region of 15q with a maximum two-point LOD score of 5.38 at D15S153 (theta = 0.00). Haplotype analysis localized the punctate PPK locus within the region defined by D15S651 and D15S988. This region overlaps by 5.06 cM with the previously reported punctate PPK region. CONCLUSIONS: This study refines a disease gene causing punctate PPK to a 5.06-cM interval at 15q22.2-15q22.31. PMID- 15888141 TI - High frequency of the 425A-->G splice-site mutation and novel mutations of the COL7A1 gene in central Europe: significance for future mutation detection strategies in dystrophic epidermolysis bullosa. AB - BACKGROUND: Mutations in the type VII collagen gene (COL7A1) are responsible for dominant and recessive forms of dystrophic epidermolysis bullosa (DEB). These mutations are usually specific for individual families; only a few cases of recurring mutations have been identified. OBJECTIVES: Forty-three unrelated Hungarian and German patients with different DEB phenotypes were screened for novel and recurrent COL7A1 mutations. METHODS: All patients were classified based on clinical and genetic findings, skin immunofluorescent antigen mapping, and electron microscopic studies. Mutation analysis was performed by amplification of genomic DNA with polymerase chain reaction using COL7A1-specific primers, heteroduplex analysis, and direct nucleotide sequencing. Restriction endonuclease digestion was used for family screening and mutation verification. Results In this group of patients, the splice-site mutation 425A-->G was observed frequently, in 11 of 86 alleles (12.8%), once in homozygous form and in nine cases in heterozygous form. One of 100 control alleles from clinically unaffected individuals also carried the mutation. We also identified three novel mutations: the 976-3C-->A splice-site mutation, and the 4929delT and 8441-15del20 deletions. CONCLUSIONS: High recurrence of the splice-site mutation 425A-->G in central European patients with DEB should be taken into account when designing COL7A1 mutation detection strategies. Reporting of three novel COL7A1 mutations in this study further emphasizes the molecular heterogeneity of DEB and provides more information for studies on genotype-phenotype correlations in different DEB subtypes. PMID- 15888142 TI - Dapsone suppresses human neutrophil superoxide production and elastase release in a calcium-dependent manner. AB - BACKGROUND: Dapsone (4,4'-diaminodiphenyl sulphone) is a powerful therapeutic tool in many skin diseases including neutrophilic dermatoses. The drug has an outstanding therapeutic efficacy against many skin diseases characterized by neutrophil-rich infiltrates; however, mechanisms of its action are poorly understood. OBJECTIVES: We investigated the effects of dapsone on respiratory and secretory functions of human neutrophils triggered by the chemotactic peptide N formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), the physiological agonist C5a, and phorbol myristate acetate (PMA). METHODS: Human neutrophils were isolated from venous blood obtained from healthy donors. We detected extracellular production of superoxide (O(2) (-)) by cytochrome C reduction assay, and intracellular production of O(2) (-) by flow cytometry. Neutrophil elastase release was measured by the cleavage of the specific elastase substrate N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide. Measurement of cytosolic free calcium concentration was performed using the calcium-reactive fluorescence probe, Fluo-3. RESULTS: Dapsone suppressed intra- and extracellular production of O(2) (-) and elastase release triggered by fMLP and C5a, but not by PMA. Both fMLP and C5a signalled the above pathways by inducing calcium influx, but PMA functions bypassed calcium influx. Dapsone was capable of antagonizing the induction of calcium influx. CONCLUSIONS: These findings suggest that one mechanism of the anti-inflammatory action of dapsone is inhibition of calcium dependent functions of neutrophils including release of tissue-damaging oxidants and proteases in the affected skin. PMID- 15888143 TI - Single genetic mutations can account for melanocytic naevi. AB - BACKGROUND: The nature of melanocytic naevi is unknown notwithstanding their considerable significance for clinician and pathologist and despite the wealth of existing knowledge about melanocyte biology. OBJECTIVES: To investigate how far a simple mutational model can explain the clinical and pathological features of melanocytic naevi, in particular their pattern of onset and frequency. METHODS: I have constructed a model of the development of the adult melanocyte population from a single stem cell. The total cutaneous melanocyte population in a human adult is already known, as well as the range of spontaneous mutation rates at a given gene site. For each cycle of mitosis during the post stem-cell expansion of the melanocyte population, I calculate the accumulated number of cells likely to be mutated at a particular (although unknown) gene site. The results are interpreted in the light of a hypothesis that each of these mutant melanocytes will go on to form a melanocytic naevus. Comparisons are made with neurofibromas, occurring in type 1 neurofibromatosis and as sporadic lesions. RESULTS: A single genetic mutation in melanocyte precursors is found to be sufficient to explain the clinical and pathological features of melanocytic naevi. CONCLUSIONS: I propose that melanocytic naevi are a consequence of single spontaneous genetic mutations which inevitably occur during the development of the adult population of cutaneous melanocytes. PMID- 15888144 TI - Acquired symmetrical dermal melanocytosis (naevus of Hori) developing after aggravated atopic dermatitis. AB - BACKGROUND: Acquired symmetrical dermal melanocytosis (ASDM) is a pigmentary disorder characterized by blue-brown macules most frequently seen on the face of young and middle-aged Asian women. ASDM developing after other skin diseases has not been previously reported. OBJECTIVES: To characterize the clinical and histopathological features of ASDM associated with atopic dermatitis (AD) and to elucidate the differences between AD-associated ASDM and idiopathic ASDM. METHODS: Sixteen patients with ASDM associated with AD were examined clinically and histopathologically and were compared with 69 patients with idiopathic ASDM. RESULTS: The AD patients associated with ASDM consisted of four men and 12 women with a mean age of 32.8 +/- 13.1 years. Most patients remembered that the pigmented macules appeared in places where refractory eczema had existed for a long time. The marked preponderance in females and the appearance in the early reproductive period were common features of AD-associated ASDM and idiopathic ASDM. AD-associated ASDM was most commonly observed on the forehead (68.8%) and on the backs of the hands (50.0%), whereas 89.9% of idiopathic ASDM was seen on the cheeks. There was no significant difference in the number of dermal melanocytes between AD-associated ASDM and idiopathic ASDM. Electron microscopic studies demonstrated many mature melanocytes and smaller numbers of immature melanocytes in the dermis. Some melanocytes were seen adjacent to mast cells. CONCLUSIONS: AD-associated ASDM does not appear to be rare in Japan. ASDM may be triggered in AD patients by sunlight exposure, some alterations in sex hormones and/or persistent cutaneous inflammation. Histamine and stem cell factor produced by mast cells may play crucial roles in the pathogenesis of AD-associated ASDM. PMID- 15888145 TI - Presence of high-risk mucosal human papillomavirus genotypes in primary melanoma and in acquired dysplastic melanocytic naevi. AB - BACKGROUND: Some studies have shown that cutaneous and mucosal melanoma biopsy specimens harbour human papillomavirus (HPV), suggesting that this virus may play a role in development and progression of the tumour. OBJECTIVES: To investigate the presence of HPV DNA and the prevalence of different high-risk mucosal HPV genotypes in primary melanoma (PM) and in acquired dysplastic melanocytic naevi (ADMN). METHODS: Fifty-one PMs from 18 men and 33 women (median age 55.5 years), 33 ADMN from 15 men and 18 women (median age 35.1 years) and 20 control skin samples from nine men and 11 women (median age 43.5 years) were studied. All diagnoses were made after histological analysis. HPV DNA analysis was made using two different polymerase chain reaction-enzyme-linked immunosorbent assay (PCR ELISA) methods, namely MY-PCR and GP-PCR. RESULTS: Using GP-PCR, mucosal HPVs were detected in 14 PMs (27%; P = 0.0166) and eight ADMN (24%; P = 0.0367), while with MY-PCR, mucosal HPVs were found in 11 PMs (22%; P = 0.04) and five ADMN (15%; P not significant). All control skin samples were negative for mucosal HPVs with both DNA amplification procedures. CONCLUSIONS: Using our PCR-ELISA methods, the detection of mucosal high-risk HPV genotypes in 24% of precursor lesions (ADMN) and in 27% of PMs adds to the body of evidence indicating a colocalization of mucosal HPV and tumoral melanocytic pathologies. PMID- 15888146 TI - Overexpression of tumour necrosis factor-alpha-converting enzyme in psoriasis. AB - BACKGROUND: Tumour necrosis factor (TNF)-alpha-converting enzyme (TACE) is a metalloproteinase-disintegrin that releases soluble TNF-alpha from cells by cleaving within the extracellular domain of membrane-bound pro-TNF-alpha. It was proposed that TNF-alpha is involved in the pathogenesis of psoriasis, and it is therefore suggested that TACE has important roles in psoriasis. However, it is unclear whether TACE is expressed in psoriatic tissue. OBJECTIVES: To clarify whether TACE is expressed in psoriatic lesions and whether expression levels of TACE mRNA are increased in lesional compared with nonlesional psoriatic skin. METHODS: Skin biopsies were obtained from patients with psoriasis. We examined the expression of TACE in psoriatic tissues using a novel real-time quantitative reverse transcriptase-polymerase chain reaction method and immunohistochemical analysis. RESULTS: There was a significant rise in the level of TACE mRNA expression in lesional psoriatic skin compared with nonlesional skin in all patients. There was a statistically significant rise in the level of TACE mRNA expression in lesional psoriatic skin compared with nonlesional skin (mean +/- SD TACE/glyceraldehyde-3-phosphate dehydrogenase ratio 0.031 +/- 0.012 vs. 0.009 +/- 0.002, P < 0.05). In lesional psoriatic skin, immunostaining with anti-TACE antibody was present throughout all layers of the epidermis. TACE immunostaining was found in the cytoplasm of keratinocytes. There was staining associated with blood vessels in the papillary dermis and perivascular inflammatory cells. In particular, mast cells showed strong staining. They contained numerous granules that were stained for TACE in the cytoplasm. CONCLUSIONS: The findings of the present study suggest that elevation of TACE mRNA in psoriatic lesions is due to many cells, particularly mast cells, that function in lesional psoriatic skin as an important source of TNF-alpha and other cytokines. PMID- 15888147 TI - Effects of drugs and anticytokine antibodies on expression of ATP2A2 and ATP2C1 in cultured normal human keratinocytes. AB - BACKGROUND: Darier's disease (DD) and Hailey-Hailey disease (HHD) are skin disorders arising, respectively, from autosomal dominant mutations in ATP2A2, encoding the sacro/endoplasmic reticulum calcium ATPase, and ATP2C1, encoding the Golgi apparatus calcium ATPase. Exposure to ultraviolet (UV) B irradiation exacerbates the skin lesions, which can be treated with corticosteroids and retinoids. OBJECTIVES: To investigate the molecular basis for DD and HHD. METHODS: We used quantitative reverse transcriptase-polymerase chain reactions to examine the effects of UVB irradiation on ATP2A2 and ATP2C1 mRNA levels in cultured normal keratinocytes. RESULTS: We observed that UVB irradiation reduced ATP2A2 and ATP2C mRNA levels. The addition of retinoids or corticosteroids to the cell culture inhibited the UVB-induced suppression of both ATP2A2 and ATP2C1 mRNA levels, and UVB-induced suppression of ATP2C1 mRNA was also inhibited by the addition of ciclosporin, tacrolimus and vitamin D(3). The addition of anti interleukin (IL)-6 antibody to the cell culture prevented the UVB-induced suppression of ATP2A2 and ATP2C1 mRNA; in contrast, the addition of anti-IL-8 antibody slightly accelerated the suppression. CONCLUSIONS: These results suggest that drugs effective for DD and HHD act by modulating ATP2A2 and ATP2C1 mRNA expression, respectively, and that the proinflammatory cytokines IL-6 and IL-8 play important roles in the regulation of ATPA2 and ATP2C1 expression in homeostasis and/or inflammation of the skin. PMID- 15888148 TI - Microarray analysis of phosphatase gene expression in human melanoma. AB - BACKGROUND: Tyrosine phosphate is abnormally elevated in malignant melanoma, and this has been interpreted to reflect the activity of oncogenic protein tyrosine kinases. However, elevation may also arise due to decreased protein tyrosine phosphatase (PTP) expression. OBJECTIVES: To survey phosphatase gene expression in melanoma cell lines, a benign naevus and normal melanocytes: we searched for downregulation of phosphatase gene expression in malignant cells that may indicate a role as melanoma suppressor genes. METHODS: Microarray analysis was used to compare gene expression for 133 phosphatase genes, comprising 39 PTPs, 16 dual-specificity phosphatases (DSPs), 47 serine/threonine phosphatases and 31 acid/alkaline and lipid-based phosphatases. Northern blotting analysis was used to study gene expression in human melanoma biopsies. RESULTS: There was decreased expression of four DSP genes (including PTEN); eight receptor PTP genes were downregulated in melanoma, among which were PTP-KAPPA and PTP-PI (consistent with our previous data). In addition, PTP-RF/LAR was downregulated in 13 of 22 metastatic melanomas. CONCLUSIONS: The expression of multiple PTP receptors is decreased in melanoma; this may be a mechanism which stimulates autonomous growth in advanced melanoma. PMID- 15888149 TI - Incidence of antiretinal antibodies in melanoma: screening of 77 serum samples from 51 patients with American Joint Committee on Cancer stage I-IV. AB - BACKGROUND: Patients with melanoma-associated retinopathy (MAR) experience different visual symptoms caused by the production of antitumoral antibodies that cross-react with retinal epitopes. Immunofluorescence assays of serum from patients with MAR on sectioned monkey or human retina characteristically reveal antibody activity located within the inner nuclear layer, with a focus of activity upon the membranes of bipolar cells. OBJECTIVES: We inquired into the association with disease of this serological abnormality by evaluating sera from patients with melanoma with no MAR-like signs or symptoms. METHODS: Groups of patients were selected with different stages of melanoma (American Joint Committee on Cancer stages I-IV). Seventy-seven serum samples from 51 patients with melanoma were examined by indirect immunohistochemical assay on sections of human retina. RESULTS: Of the 77 serum samples, 53 were found to contain antibodies reactive with various components of retina. Eight were from 17 sera from patients in stage I or II, 14 were from 23 sera from patients in stage III, and 31 were from 37 sera from patients in stage IV. Statistical analysis revealed a correlation between antibody activity and the stage of disease, with a higher percentage of antibody activity in advanced stages (P = 0.002). CONCLUSIONS: The presence of antiretinal antibodies in patients with melanoma without ocular symptoms appears to be more common than previously suspected. Antibody activity similar to that ascribed to the MAR syndrome appears in some patients with melanoma who have no MAR-like retinopathy. Follow-up studies will determine if patients with antiretinal antibodies go on to develop MAR and if staining intensity and staining patterns change over the course of the disease. PMID- 15888150 TI - Imiquimod 5% cream for the treatment of superficial basal cell carcinoma: results from a randomized vehicle-controlled phase III study in Europe. AB - BACKGROUND: Imiquimod is an immune response modifier that acts through toll-like receptor 7 to induce cytokine production and a subsequent innate and adaptive cell-mediated immune response. Clinical studies have demonstrated clinical and histological clearance of superficial basal cell carcinoma (sBCC) after treatment with imiquimod 5% cream. OBJECTIVES: To evaluate the safety and clinical efficacy of imiquimod (Aldaratrade mark; 3M Pharmaceuticals, St Paul, MN, U.S.A.) 5% cream for the treatment of sBCC in a multicentre, randomized, parallel, vehicle controlled, double-blind, phase III clinical study conducted at 26 centres in Europe. METHODS: Subjects who had at least one histologically confirmed sBCC tumour were randomized to apply imiquimod or vehicle cream to the target tumour once daily, seven times per week (7 x/week) for 6 weeks. The target tumour location was identified with an indelible ink mark before treatment initiation. The treated tumour site was clinically assessed for treatment response at 12 weeks post-treatment and was then excised for histological evaluation. Efficacy assessments included the composite response rates (proportion of subjects with clinical and histological clearance) and response rates solely based on histology (proportion of subjects with histological clearance). Safety assessments, which included adverse events and scoring of local skin reactions (LSRs), were carried out throughout the study. RESULTS: In total, 166 subjects were enrolled in this study. For the intent-to-treat dataset, there was a statistically significant difference between imiquimod and vehicle groups for both composite clearance rates (clinical and histological assessments) and histological clearance rates. Composite clearance was demonstrated in 77% and 6% of subjects treated with imiquimod and vehicle cream, respectively. Histological clearance was demonstrated in 80% and 6% of subjects treated with imiquimod and vehicle cream, respectively. The most frequently reported safety findings were investigator assessed LSRs and spontaneous reports by subjects of application site reactions, which occurred more frequently in the imiquimod group than in the vehicle group. CONCLUSIONS: Imiquimod 5% cream administered 7 x/week for 6 weeks is a safe and effective treatment for sBCC when compared with vehicle cream. PMID- 15888151 TI - High plasma proteasome levels are detected in patients with metastatic malignant melanoma. AB - BACKGROUND: Proteasomes, nonlysosomal proteolytic structures, are implicated in cell growth and differentiation. An abnormal expression has been described in haematopoietic malignancies and in some solid tumours. OBJECTIVES: To study the plasma proteasome levels in patients with malignant melanoma (MM) using an enzyme linked immunosorbent assay (ELISA) technique, and to compare them with the values obtained in a normal population and in patients with severe psoriasis or chronic idiopathic urticaria (CIU). METHODS: Plasma proteasome level was measured using a sandwich ELISA test in normal donors (n = 14), and in patients with stage I/II (n = 13), stage III (n = 6) and stage IV (n = 10) MM, severe psoriasis (n = 13) and CIU (n = 6). Tissue proteasome expression was also detected by immunohistology using a monoclonal antibody in paraffin-embedded samples of normal tissue, psoriasis skin and MM. RESULTS: In normal donors, mean +/- SEM plasma proteasome concentration was 2138 +/- 221 ng mL(-1). Patients with stages III and IV MM exhibited a significantly higher value (3373 +/- 470 ng mL(-1) and 8931 +/- 1232 ng mL(-1), respectively). Values in patients with stage I/II MM and CIU were not significantly different from those in normal volunteers. Patients with severe psoriasis also exhibited increased values (3398 +/- 374 ng mL(-1)) but to a lesser extent than in patients with stage IV MM. There was a significant correlation of proteasome levels with serum lactate dehydrogenase in the MM group. Tissue expression as demonstrated by immunohistochemistry paralleled these findings. The strongest expression was seen on MM slides and to a lesser extent in psoriasis samples, the weakest expression being observed in normal skin. CONCLUSIONS: Proteasomes are strongly expressed in cutaneous MM; high levels of circulating proteasomes are detected in patients with metastatic MM with a high melanoma burden, and at a lesser extent in psoriatic patients, which suggests proteasomes represent a marker more of nonspecific inflammation than of early cancer. PMID- 15888152 TI - Infliximab treatment results in significant improvement in the quality of life of patients with severe psoriasis: a double-blind placebo-controlled trial. AB - BACKGROUND: Psoriasis is a chronic disease that significantly diminishes the health-related quality of life (HRQOL). Infliximab is a chimeric, tumour necrosis factor alpha monoclonal antibody that has been shown to improve the signs and symptoms of plaque psoriasis. OBJECTIVES: The objective of this study was to evaluate the effect of infliximab induction therapy on the HRQOL of patients with severe plaque psoriasis. METHODS: In this double-blind, placebo-controlled trial, 249 patients were randomly assigned to receive intravenous infusions of 3 or 5 mg kg(-1) of infliximab or placebo and were treated at weeks 0, 2 and 6. Patients completed the Dermatology Life Quality Index (DLQI) at baseline and week 10. RESULTS: Infliximab induction therapy resulted in a substantial improvement in HRQOL. At week 10, patients in the infliximab 3- and 5-mg kg(-1) groups showed a median percentage improvement in DLQI scores of 84.0% and 91.0%, respectively, compared with 0% in the placebo group (P < 0.001). The median decrease from baseline in DLQI score at week 10 was 8.0 and 10.0 for the 3 and 5 mg kg(-1) infliximab groups, respectively, compared with 0 in the placebo group (P < 0.001). Thirty-three per cent and 40% of patients in the 3 and 5 mg kg(-1) infliximab groups, respectively, had a DLQI score of 0 at week 10, compared with 2% in the placebo group (P < 0.001). There was a strong correlation between the percentage change from baseline at week 10 in Psoriasis Area and Severity Index (PASI) scores and the percentage change in DLQI scores during the same period (Spearman's correlation, 0.61, P < 0.001). When the infliximab and placebo treatment groups were combined, patients with at least 75% improvement in PASI scores between baseline and week 10 had a greater mean improvement in DLQI scores (81%) than those with 50-75% improvement in PASI during the same period (60%). CONCLUSIONS: Infliximab induction therapy resulted in significant improvement in HRQOL in patients with severe psoriasis. PMID- 15888153 TI - Human papillomavirus-associated plantar epidermoid cyst related to epidermoid metaplasia of the eccrine duct epithelium: a combined histological, immunohistochemical, DNA-DNA in situ hybridization and three-dimensional reconstruction analysis. AB - BACKGROUND: We recently proposed that certain palmoplantar epidermoid cysts may be related to eccrine ducts and that human papillomavirus (HPV) 60 may play a role in their pathomechanism. However, the origin of palmoplantar epidermoid cysts is still controversial. OBJECTIVES: To examine the contribution of eccrine ducts and HPV 60 in the development of epidermoid cysts. METHODS: Five epidermoid cysts and four ridged warts that had developed on the soles of a patient were studied histologically, immunohistochemically and by DNA-DNA in situ hybridization. Using serial sections obtained from its entire body, a three dimensional reconstruction (3DR) analysis was performed on the smallest cyst to analyse the relationship between the epidermoid cyst, eccrine duct and the overlying epidermis. RESULTS: Histological and DNA-DNA in situ hybridization analyses demonstrated both homogeneous intracytoplasmic inclusion bodies pathognomonic for HPV 60 infection and HPV 60 DNA sequences not only in all of the epidermoid cysts and ridged warts but also in the acrosyringeal portion of an eccrine duct, with the dermal portion of which the smallest cyst had been revealed to connect by 3DR analysis. However, immunohistochemical analyses using antibodies against human carcinoembryonic antigen (CEA), involucrin and several cytokeratins (CKs) revealed that the immunoreactivity of the cyst was not identical to that of the eccrine dermal duct but was identical to that of suprabasal layers of the epidermis. CONCLUSIONS: It was clearly demonstrated that an HPV 60-associated epidermoid cyst with immunoreactivities for CEA, involucrin and CKs which were identical to those of the epidermis connected with the eccrine dermal duct, supporting the idea that certain palmoplantar epidermoid cysts may develop following the epidermoid metaplasia of eccrine ducts with HPV 60 infection. PMID- 15888154 TI - The usefulness of skin tests to prove drug hypersensitivity. AB - BACKGROUND: Suspected drug hypersensitivity is common. Only a minority of cutaneous adverse drug reactions (CADRs) are allergic in origin and will reappear after the next exposure. Methods to confirm suspected CADRs are needed and skin testing could serve as one possibility. OBJECTIVES: To analyse the usefulness of skin tests in revealing drug allergy. The relevance of skin test results was evaluated with drug provocation studies. METHODS: During 1989-2001, 947 patients with a history of suspected CADR were examined with skin tests including patch tests (PTs) (826 patients), skin prick tests (SPTs) (935 patients) and photopatch tests (12 patients). The occurrence of positive and negative test reactions to different drugs was correlated with clinical history. Drug provocation was carried out in 246 patients. RESULTS: Antimicrobial drugs were suspected and tested most often. A positive PT reaction to one or more drug was seen in 89 of 826 (10.8%), most often to beta-lactams, clindamycin and trimethoprim. A positive SPT reaction was seen in 10 of 935 (1.1%) patients. Challenge was carried out in 17 patients with positive skin test results. Thirteen of 16 (81.2%) PT positives developed exanthema, three remained negative and one SPT-positive patient developed urticaria. Among skin test negatives, 207 of 229 (90.4%) challenges were negative and 22 of 229 (9.6%) were positive, 12 with exanthema, three with fixed drug eruptions and seven with urticaria. CONCLUSIONS: Skin testing, especially the PT, was a useful screening method to find a cause of CADR if the reaction was exanthema and if antimicrobial, cardiovascular or antiepileptic drugs were suspected. The SPT detected occasional positives with antimicrobials. In cases of fixed drug eruption, PTs performed at the earlier reaction site were useful. When skin tests are negative or dubious, oral challenge should be carried out to confirm the association. PMID- 15888155 TI - Fifteen-year follow-up of hand eczema: persistence and consequences. AB - BACKGROUND: Hand eczema is a skin disease often with a long-lasting and relapsing course. The long-term prognosis in the general population is unknown. OBJECTIVES: The aims were to examine the extent to which hand eczema had persisted and the medicosocial consequences of the disease. METHODS: In a 15-year follow-up of hand eczema, patients diagnosed in a previous population-based study were sent a questionnaire with 20 questions concerning the persistence and course of the disease, and its occupational and medicosocial consequences. RESULTS: Addresses were available for 1115 persons, of whom 868 answered the questionnaire. Sixty six per cent of the respondents reported periods of hand eczema and 44% reported symptoms during the previous year, with no sex difference. Twelve per cent reported continuous eczema. However, 74% of those reporting symptoms considered that their hand eczema had improved; of these more were women than men (78% vs. 66%, P < 0.01). Twenty people, 3% of those who were gainfully employed in 1983, reported a change to another occupation because of their hand eczema, 15 of these reporting improvement after the job change. A considerable need for medical consultation was reported, as was the influence on psychosocial functions among those who had eczema the previous year, e.g. sleep disturbances (36%) and hampered leisure activities (72%). Job changes related to hand eczema and psychosocial impairment were also reported by individuals who had not sought medical help for their hand eczema. CONCLUSIONS: This study demonstrates a variable and poor long-term prognosis for hand eczema in the general population. One-third sought medical care during follow-up, while the vast majority with ongoing hand eczema experienced negative psychosocial consequences. For about 5%, the hand eczema gave far-reaching consequences including long sick-leave periods, sick pension and changes of occupation. PMID- 15888156 TI - Optimal weekly frequency of 308-nm excimer laser treatment in vitiligo patients. AB - BACKGROUND: Recently the beneficial effect of excimer laser treatment has been reported for patients with vitiligo. The influence of treatment frequency on this effect is not clear. OBJECTIVES: To determine the optimal frequency of 308-nm excimer laser therapy for vitiligo. METHODS: In this prospective, university based hospital study over 12 weeks we enrolled 14 patients. Each had at least three stable vitiligo lesions in the same body area. The three stable vitiligo lesions in each subject were randomly assigned to receive excimer laser treatment once (1 x), twice (2 x) and three times (3 x) weekly, respectively. The initial ultraviolet (UV) dose was 50 mJ cm(-2) less than the 308-nm minimal erythematous dose in vitiligo skin. The UV dose was increased at each treatment session according to the erythematous response to the previous treatment. RESULTS: Thirteen subjects were treated for at least 6 weeks; seven were treated for all 12 weeks. At 6 weeks, the repigmentation rates for treated lesions were 8% (1/13) after 1 x weekly treatment, 23% (3/13) after 2 x weekly treatment and 62% (8/13) after 3 x weekly treatment (P = 0.0134; 3 x vs. 1 x weekly); at 12 weeks, these rates were 46% (6/13), 62% (8/13) and 69% (9/13), respectively (P = NS; 3 x vs. 1 x weekly). Repigmentation initiation correlated with treatment number, regardless of frequency (P = NS). As shown by Kaplan-Meier analysis, repigmentation occurred earliest in the most frequently treated lesions (P = 0.0336). At 12 weeks, the projected repigmentation rates for 1 x, 2 x and 3 x weekly treatment approached each other (60%, 79% and 82%, respectively); the mean repigmentation grades (on a scale of 0-5) for 1 x, 2 x and 3 x weekly treatment were 1.7, 2.4 and 3.3, respectively (P = 0.018; 3 x vs. 1 x weekly). Laser-induced repigmentation persisted in most cases over the entire follow-up of 12 months after the end of treatment. CONCLUSIONS: 308-nm excimer laser therapy is effective against vitiligo. Although repigmentation occurs fastest with 3 x weekly treatment, the ultimate repigmentation initiation seems to depend entirely on the total number of treatments, not their frequency. However, treatment periods of more than 12 weeks may be necessary to obtain a satisfactory clinical repigmentation, particularly when vitiligo lesions are treated only 1 x or 2 x compared with 3 x weekly. PMID- 15888157 TI - A randomized controlled trial of laser treatment among hirsute women with polycystic ovary syndrome. AB - BACKGROUND: Facial hirsutism is one of the characteristic features of polycystic ovary syndrome (PCOS), and this can lead to high levels of depression and anxiety. OBJECTIVES: To evaluate the impact of laser treatment on the severity of facial hirsutism and on psychological morbidity in women with PCOS. METHODS: A randomized controlled trial of five high-fluence treatments (intervention) vs. five low-fluence treatments (control) was performed over 6 months in a National Health Service teaching hospital. Subjects were 88 women with facial hirsutism due to PCOS recruited from hospital outpatient clinics and a patient support group in 2001-2002. The main outcomes were self-reported severity of facial hair (measured on a scale of 1-10), depression, anxiety (measured on the Hospital Anxiety and Depression Scale) and quality of life (measured on the WHOQOL-BREF). RESULTS: Self-reported severity of facial hair in the intervention group (n = 51) fell from 7.3 to 3.6 over the 6-month study period; for the control group (n = 37) the corresponding scores were 7.1 and 6.1. The change was significantly greater in the intervention group [ancova F((1,83)) = 24.5, P < 0.05]. Self reported time spent on hair removal declined from 112 to 21 min per week in the intervention group and from 92 to 56 min in the control group [F((1,80)) = 10.2, P or=0.1%) was virucidal against cell-free and cell-associated HIV-1 in breast milk. SDS could be substantially removed from breast milk, without recovery of viral infectivity. Viral load in artificially infected milk was reduced to undetectable levels after treatment with 0.1% SDS. SDS was virucidal against HIV 1 in human milk and could be removed from breast milk if necessary. Milk was not infectious after SDS removal. CONCLUSION: The proposed treatment concentrations are within reported safe limits for ingestion of SDS by children of 1 g/kg/day. Therefore, use of alkyl sulfate microbicides, such as SDS, to treat HIV1-infected breast milk may be a novel alternative to help prevent/reduce transmission of HIV 1 through breastfeeding. PMID- 15888211 TI - Effects of naloxone on respiratory sensation before and after a removal of severe respiratory stress. AB - Severe respiratory stress causes dyspnea, and a sudden release of this stress frequently accompanies a euphoric sensation. We hypothesized that acute severe respiratory stress may result in an elaboration of endogenous opioids within the central nervous system, and that these opioids may play significant roles in relieving dyspnea and generating euphoric sensation after a sudden removal of the stress. To test this hypothesis, we examined the effects of naloxone (0.04 mg/kg, I.V.) and the placebo (normal saline) on changes in respiratory sensation before and after the release of severe respiratory stress in a double-blind, randomized, crossover study in 14 healthy adults. Acute severe respiratory stress was induced by loaded breathing with a combination of resistive loading and hypercapnia. The subjects rated their changes in sensation by using a bidirectional visual analogue scale. Naloxone pretreatment affected neither the ventilation nor the development of dyspneic sensation during loaded breathing. Naloxone pretreatment only slightly attentuated the euphoric sensation developed after the release of severe respiratory stress. These findings suggest a small role of opioids in relieving dyspnea and in generating euphoria before and after a sudden removal of stress. PMID- 15888212 TI - Peas and primroses. PMID- 15888213 TI - Developing academic-practice partnerships to enhance the integration of genomics into public health. AB - The sequencing of the human genome has provided tools to gain a better understanding of the role of genes and their interaction with environmental factors in the development of disease. However, much work remains in translating discoveries into new opportunities for disease prevention and health promotion. Both public health academia and practice have important roles to play in bridging the gap between the growth in knowledge stemming from the Human Genome Project and its application in public health. Recognizing this, the Centers for Disease Control and Prevention, through the Association of Schools of Public Health, established Centers for Genomics and Public Health at three schools of public health in 2001: the University of Michigan, the University of North Carolina, and the University of Washington. This paper describes the experience of the University of Washington Center for Genomics and Public Health in forging partnerships with public health practitioners to translate genomic advances into public health practice. PMID- 15888214 TI - Using evidence-based community and behavioral interventions to prevent skin cancer: opportunities and challenges for public health practice. PMID- 15888215 TI - Blazing a trail: a public health research agenda in genomics and chronic disease. PMID- 15888216 TI - Genomics and the prevention and control of common chronic diseases: emerging priorities for public health action. PMID- 15888217 TI - How do we ensure the quality of the public health workforce? PMID- 15888219 TI - Review of state Comprehensive Cancer Control plans for genomics content. AB - INTRODUCTION: The goals of this study were to determine U.S. states with Comprehensive Cancer Control plans that include genomics in some capacity and to review successes with and barriers to implementation of genomics-related cancer control initiatives. METHODS: This study was conducted in two phases. Phase one included a content analysis of written state Comprehensive Cancer Control plans (n = 30) for terms related to genomics, or "genomic components" (n = 18). The second phase involved telephone interviews with the Comprehensive Cancer Control plan coordinators in states with plans that contained genomic components (n = 16). The interview was designed to gather more detailed information about the genomics-related initiatives within the state's Comprehensive Cancer Control plan and the successes with and barriers to plan implementation, as defined by each state. RESULTS: Eighteen of the 30 Comprehensive Cancer Control plans analyzed contained genomics components. We noted a large variability among these 18 plans in the types of genomics components included. Nine (56%) of the 16 states interviewed had begun to implement the genomics components in their plan. Most states emphasized educating health care providers and the public about the role of genomics in cancer control. Many states consider awareness of family history to be an important aspect of their Comprehensive Cancer Control plan. Approximately 67% of states with family history components in their plans had begun to implement these goals. Virtually all states reported they would benefit from additional training in cancer genetics and general public health genomics. CONCLUSION: The number of states incorporating genomics into their Comprehensive Cancer Control plans is increasing. Family history is a public health application of genomics that could be implemented more fully into Comprehensive Cancer Control plans. PMID- 15888220 TI - Adherence to oral hypoglycemic agents in Hawaii. AB - INTRODUCTION: Adherence to oral hypoglycemic agents is essential to reducing the poor health outcomes of populations at high risk for developing diabetes and its chronic complications. The goal of this study was to identify characteristics of patients in Hawaii least likely to adhere to oral hypoglycemic agents. METHODS: This retrospective administrative data analysis included prescription refill claims for oral hypoglycemic agents from January 1, 1999, through June 30, 2003 (n = 20,685). Multivariate logistic regression analysis was used to examine the relationship between adherence and patient characteristics. RESULTS: Adherence was found to be strongly associated with age and ethnicity. Relative to the age subset 55 to 64 years, adherence increased as age increased, reaching a peak at age 74 (odds ratio [OR] 1.1; 95% confidence interval [CI], 1.0-1.20). Past the age of 85, adherence declined (OR 0.90; 95% CI, 0.82-0.98). Relative to white patients, the odds ratio of adherence was highest for Japanese patients (OR 1.20; 95% CI, 1.0-1.30) and lowest for Filipino patients (OR 0.78; 95% CI, 0.68-0.90). Gender was not associated with adherence. CONCLUSION: Differences in adherence to oral hypoglycemic agents were found to be related to ethnicity and age. Adherence was found to be lowest in younger patients and Filipino patients. This is a significant finding considering that younger diabetic patients have been shown to have the poorest glycemic control and worst health outcomes. Although the literature on adherence to oral hypoglycemic agents and health outcomes in Filipino patients is limited, studies support an increased risk for developing diabetes in this group. This information can be used to target younger patients and Filipino patients to improve their adherence to oral hypoglycemic agents. PMID- 15888221 TI - Using the State Plan Index to evaluate the quality of state plans to prevent obesity and other chronic diseases. AB - INTRODUCTION: Implicit in public health planning models is the assumption that good public health plans lead to good programs, and good programs lead to desired health outcomes. Despite considerable resources that are devoted to developing plans, public health agencies and organizations have lacked a tool for evaluating the finished product of their planning efforts -- the written plan itself -- as an important indicator of progress. To address the need for an instrument to assess the quality of state plans designed to prevent and control chronic diseases, we created and tested the State Plan Index and used it to evaluate the quality of nine state plans aimed at preventing and reducing obesity. METHODS: The State Plan Index was developed under the auspices of the Centers for Disease Control and Prevention (CDC) in collaboration with public health experts in federal, state, and academic settings. The State Plan Index included 55 items related to plan quality arranged into nine components. Each item was rated on a Likert scale from 0 to 5, with 5 being the highest rating. Each plan also received a separate overall plan quality score using the same scale. Each state plan was evaluated by four or five raters using the State Plan Index. For each plan, the 55 items were averaged to calculate an item average score, and a subscore was calculated for each State Plan Index component. Finally, five states also self-rated their own plans (self score). RESULTS: The mean item average score for all plans was 2.4 out of 5.0. The range of item average scores was 1.0 to 3.0. The component of the State Plan Index with the highest mean component score (3.3) was Presentation of Epidemiologic Data on Disease Burden. The components with the lowest component scores were Resources for Plan Implementation (0.7); Integration of Obesity Efforts with Other Chronic Disease Efforts (1.7); and Program Evaluation (2.0). Plan quality was rated higher when based on the single overall plan quality score assigned by raters. In addition, self scores were consistently and substantially higher than rater-assigned scores. CONCLUSION: Evaluation of plans early in the life of programs can be used to strengthen existing programs and to guide programs newly engaged in chronic disease prevention planning. The CDC has used the State Plan Index evaluation results to guide technical assistance, plan training sessions, and enhance communication with state staff about plan content, quality, and public health approach. Some state program directors self-evaluated their obesity draft plan and used the evaluation results to strengthen their planning process and to guide plan revisions. Other states have adapted the State Plan Index as a framework for new planning efforts to prevent obesity as well as other chronic diseases. PMID- 15888222 TI - Development of a brief survey on colon cancer screening knowledge and attitudes among veterans. AB - INTRODUCTION: Poor knowledge of and negative attitudes toward available screening tests may account in part for colorectal cancer screening rates being the lowest among 17 quality measures reported for the Department of Veterans Affairs health care system, the largest integrated health system in the United States. The purpose of this study was to develop a brief assessment tool to evaluate knowledge and attitudes among veterans toward colorectal cancer screening options. METHODS: A 44-item questionnaire was developed to assess knowledge, attitudes, and beliefs about colorectal cancer and screening and was then administered as part of an ongoing randomized controlled trial among 388 veterans receiving care in a general medicine clinic. Sixteen candidate items on colorectal cancer knowledge, attitudes, and beliefs were selected for further evaluation using principal components analysis. Two sets of items were then further analyzed. RESULTS: Because the Cronbach alpha for beliefs was low (alpha = 0.06), the beliefs subscale was deleted from further consideration. The final scale consisted of seven items: a four-item attitude subscale (alpha = 0.73) and a three-item knowledge subscale (alpha = 0.59). Twelve-month follow-up data were used to evaluate predictive validity; improved knowledge and attitudes were significantly associated with completion of flexible sigmoidoscopy (P = .004) and completion of either flexible sigmoidoscopy or colonoscopy (P = .02). CONCLUSION: The two-factor scale offers a parsimonious and reliable measure of colorectal cancer screening knowledge and attitudes among veterans. This colorectal Cancer Screening Survey (CSS) may especially be useful as an evaluative tool in developing and testing of interventions designed to improve screening rates within this population. PMID- 15888223 TI - Secular trends in age at menarche, smoking, and oral contraceptive use among Israeli girls. AB - INTRODUCTION: The improved nutrition and socioeconomic status of the population in industrialized countries has resulted in a decrease in the mean age at menarche. This trend raises the question of whether cigarette smoking and the use of oral contraceptives, health behaviors often adopted during adolescence, may also be starting at a younger age. Cigarette smoking and use of oral contraceptives are a public health concern because they pose an increased risk for development of chronic diseases, particularly in combination. This study was designed to identify secular trends in age at menarche, at first cigarette, and at first use of oral contraceptives among a large population-based sample of young Israeli women and to assess whether these trends are associated with sociodemographic factors. METHODS: A systematic, population-based survey used data obtained from female recruits to the Israel Defense Force from 1986 to 2000. During the study period, 11,392 questionnaires were collected from Jewish women aged 18 to 19 years. Participants were interviewed concerning geographic origin and level of education, father's geographic origin and level of education, current smoking status, use of oral contraceptives, and recalled age at first menstruation, first cigarette, and first use of oral contraceptives. RESULTS: Reported mean age (-/+ D) at menarche showed a monotonic trend of decreasing over time, from 13.41 (-/+ .30) years for women born before 1970 to 13.03 (-/+ 1.28) years for those born after 1978 (P < .001). Women born after 1978 were twice as likely to experience menarche by the age of 11 as those born prior to 1970 (odds ratio 2.0; 95% confidence interval, 1.41-2.82). Significant trends toward younger age at first use were observed for cigarettes and oral contraceptives. CONCLUSION: The trends of earlier age at menarche, first cigarette, and first use of oral contraceptives suggest health behaviors among young women that may herald increased chronic disease morbidity in the future. These trends indicate the need for further investigation and preventive measures aimed at this population. PMID- 15888224 TI - Knowledge and perceptions of diabetes in an Appalachian population. AB - INTRODUCTION: Qualitative research on knowledge and perceptions of diabetes is limited in the Appalachian region, where social, economic, and behavioral risk factors put many individuals at high risk for diabetes. The aim of this study was to gain a culturally informed understanding of diabetes in the Appalachian region by 1) determining cultural knowledge, beliefs, and attitudes of diabetes among those who live in the region; 2) identifying concerns and barriers to care for those with diabetes; and 3) determining the barriers and facilitators to developing interventions for the prevention and early detection of diabetes in Appalachia. METHODS: Thirteen focus groups were conducted in 16 counties in West Virginia in 1999. Seven of the groups were composed of persons with diabetes (n = 61), and six were composed of community members without diabetes (n = 40). Participants included 73 women and 28 men (n = 101). RESULTS: Findings show that among this population there is lack of knowledge about diabetes before and after diagnosis and little perception that a risk of diabetes exists (unless there is a family history of diabetes). Social interactions are negatively affected by having diabetes, and cultural and economic barriers to early detection and care create obstacles to the early detection of diabetes and education of those diagnosed. CONCLUSION: Public health education and community-level interventions for primary prevention of diabetes in addition to behavior change to improve the management of diabetes are needed to reduce the health disparities related to diabetes in West Virginia. PMID- 15888225 TI - Rural community knowledge of stroke warning signs and risk factors. AB - INTRODUCTION: Rapid identification and treatment of ischemic stroke can lead to improved patient outcomes. Public education campaigns in selected communities have helped to increase knowledge about stroke, but most data represent large metropolitan centers working with academic institutions. Much less is known about knowledge of stroke among residents in rural communities. METHODS: In 2004, 800 adults aged 45 years and older from two Montana counties participated in a telephone survey using unaided questions to assess awareness of stroke warning signs and risk factors. The survey also asked respondents if they had a history of atrial fibrillation, diabetes, high blood pressure, high cholesterol, smoking, heart disease, or stroke. RESULTS: More than 70% of survey participants were able to correctly report two or more warning signs for stroke: numbness on any side of the face/body (45%) and speech difficulties (38%) were reported most frequently. More than 45% were able to correctly report two or more stroke risk factors: smoking (50%) and high blood pressure (44%) were reported most frequently. Respondents aged 45 to 64 years (odds ratio [OR] 2.44; 95% confidence interval [CI], 1.78-3.46), women (OR 2.02; 95% CI, 1.46-2.80), those with 12 or more years of education (OR 1.96; 95% CI, 1.08-3.56), and those with high cholesterol (OR 1.68; 95% CI, 1.17-2.42) were more likely to correctly identify two or more warning signs compared with respondents without these characteristics. Women (OR 1.48; 95% CI, 1.07-2.05) and respondents aged 45 to 64 years (OR 1.35; 95% CI, 1.01-1.81) were also more likely to correctly identify two or more stroke risk factors compared with men and older respondents. CONCLUSION: Residents of two rural counties were generally aware of stroke warning signs, but their knowledge of stroke risk factors was limited. PMID- 15888226 TI - State Plan Index: a tool for assessing the quality of state public health plans. AB - INTRODUCTION: The State Plan Index is an evaluation instrument that uses a Likert scale to assess 60 indicators of the quality of state public health plans. The State Plan Index was needed to enable evaluation of plans that were developed using a variety of public health planning models. METHODS: Federal, state, and academic partners participated in developing and testing the instrument. The authors conducted a literature review, interviews with experts, and several rounds of formative evaluation to assess item inclusion, coverage, weighting, organization of items, and content validity. In two rounds of field testing, public health practitioners at the federal and state levels rated 10 state public health plans for obesity prevention. RESULTS: Field-test raters took an average of two hours to rate a plan and indicated that the State Plan Index was "easy to use," "comprehensive," and "fair." Mean Cronbach alpha for components of the State Plan Index was 0.88 (median 0.93). Component scores among the 10 plans rated ranged from 0.2 to 4.8, indicating that raters made distinctions in quality among the components and the plans they rated. Correlations between component scores and overall scores were statistically significant (P < .001), except for one component. CONCLUSION: Public health professionals at the federal and state levels found the State Plan Index to be a useful tool for evaluating public health plans that were developed by states using various planning approaches. After the field tests, state staff reported adapting the State Plan Index for use as a planning tool, an evaluation tool for local plans, and a self-assessment tool for drafts of state plans. In addition, the State Plan Index can be revised easily for use in other chronic disease areas. PMID- 15888227 TI - Anthropometric changes using a walking intervention in African American breast cancer survivors: a pilot study. AB - INTRODUCTION: African American women exhibit a higher mortality rate from breast cancer than do white women. African American women are more likely to gain weight at diagnosis, which may increase their risk of cancer recurrence and comorbidities. Physical activity has been shown to decrease body mass index and improve quality of life in cancer survivors. This study was designed to evaluate the feasibility and impact of a community-based exercise intervention in African American breast cancer survivors. METHODS: A theory-based eight-week community intervention using pedometers with scheduling, goal setting, and self-assessment was tested in a convenience sample of African American breast cancer survivors (n = 24). Data were collected at three time points to examine changes in steps walked per day, body mass index, and other anthropometric measures, attitudes, and demographic variables. RESULTS: Statistically significant increases in steps walked per day and attitude toward exercise as well as significant decreases in body mass index, body weight, percentage of body fat, and waist, hip, and forearm circumferences, as well as blood pressure, were reported from baseline to immediate post-intervention. Positive changes were retained or improved further at three-month follow-up except for attitude toward exercise. Participant retention rate during eight-week intervention was 92%. CONCLUSION: Increasing walking for exercise, without making other changes, can improve body mass index, anthropometric measures, and attitudes, which are associated with improved quality of life and reduced risk of cancer recurrence. The high participant retention rate, along with significant study outcomes, demonstrate that among this sample of African American breast cancer survivors, participants were motivated to improve their exercise habits. PMID- 15888228 TI - Prevalence of physical activity in the United States: Behavioral Risk Factor Surveillance System, 2001. AB - INTRODUCTION: The health benefits of regular cardiovascular exercise are well known. Such exercise, however, has traditionally been defined as vigorous physical activity, such as jogging, swimming, or aerobic dance. Exercise of moderate intensity also promotes health, and many U.S. adults may be experiencing the health benefits of exercise through lifestyle activities of moderate intensity, such as yard work, housework, or walking for transportation. Until recently, public health surveillance systems have not included assessments of this type of physical activity, focusing on exercise of vigorous intensity. We used an enhanced surveillance tool to describe the prevalence and amount of both moderate-intensity and vigorous-intensity physical activity among U.S. adults. METHODS: We analyzed data from the 2001 Behavioral Risk Factor Surveillance System, a state-based, random-digit-dialed telephone survey administered to U.S. adults aged 18 years and older (n = 82,834 men and 120,286 women). Physical activity behavior was assessed using questions designed to quantify the frequency of participation in moderate- or vigorous-intensity physical activities performed during leisure time or for household chores and transportation. RESULTS: Overall, 45% of adults (48% of men and 43% of women) were active at recommended levels during nonworking hours (at least 30 minutes five or more days per week in moderate-intensity activities, equivalent to brisk walking, or at least 20 minutes three or more days per week in vigorous activities, equivalent to running, heavy yard work, or aerobic dance). Less than 16% of adults (15% of men and 17% of women) reported no moderate or vigorous activity in a usual week. CONCLUSION: Integrating surveillance of lifestyle activities into national systems is possible, and doing so may provide a more accurate representation of the prevalence of recommended levels of physical activity. These results, however, suggest that the majority of U.S. adults are not active at levels associated with the promotion and maintenance of health. PMID- 15888229 TI - Adherence to heart-healthy behaviors in a sample of the U.S. population. AB - INTRODUCTION: Following national recommendations for physical activity, diet, and nonsmoking can reduce both incident and recurrent coronary heart disease. Prevalence data about combinations of behaviors are lacking. This study describes the prevalence of full adherence to national recommendations for physical activity, fruit and vegetable consumption, and nonsmoking among individuals with and without coronary heart disease and examines characteristics associated with full adherence. METHODS: We performed a cross-sectional analysis of data from the 2000 Behavioral Risk Factor Surveillance System, a national population-based survey. We included respondents to the cardiovascular disease module and excluded individuals with poor physical health or activity limitations. RESULTS: Subjects were most adherent to smoking recommendations (approximately 80%) and less adherent to fruit and vegetable consumption and physical activity (approximately 20% for both). Only 5% of those without coronary heart disease and 7% of those with coronary heart disease were adherent to all three behaviors (P < .01). Among those without a history of coronary heart disease, female sex (odds ratio [OR] 1.47; 95% confidence interval [CI], 1.23-1.76), highest age quintile (OR 1.67; 95% CI, 1.28-2.19), more education (OR 2.48; 95% CI, 1.69-3.64), and more income (OR 1.19; 95% CI, 1.04-1.36) were associated with full adherence. Among those with coronary heart disease, mid-age quintile (OR 3.79; 95% CI, 1.35-10.68), good general health (OR 2.05; 95% CI, 1.07-3.94), and more income (OR 1.51; 95% CI, 1.06-2.16) were associated with full adherence. CONCLUSION: These data demonstrate the lack of a heart-healthy lifestyle among a sample of U.S. adults with and without coronary heart disease. Full adherence to combined behaviors is far below adherence to any of the individual behaviors. PMID- 15888230 TI - Family history, diabetes, and other demographic and risk factors among participants of the National Health and Nutrition Examination Survey 1999-2002. AB - INTRODUCTION: Family history of diabetes has been recognized as an important risk factor of the disease. Family medical history represents valuable genomic information because it characterizes the combined interactions between environmental, behavioral, and genetic factors. This study examined the strength and effect of having a family history of diabetes on the prevalence of self reported, previously diagnosed diabetes among adult participants of the National Health and Nutrition Examination Survey 199-2002. METHODS: The study population included data from 10,283 participants aged 20 years and older. Gender, age, race/ethnicity, poverty income ratio, education level, body mass index, and family history of diabetes were examined in relation to diabetes status. Diabetes prevalence estimates and odds ratios of diabetes were calculated based on family history and other factors. RESULTS: The prevalence of diabetes among individuals who have a first-degree relative with diabetes (14.3%) was significantly higher than that of individuals without a family history (3.2%), corresponding to a crude odds ratio of five. Both prevalence and odds ratio estimates significantly increased with the number of relatives affected with diabetes. Family history was also associated with several demographic and risk factors. CONCLUSION: Family history of diabetes was shown to be a significant predictor of diabetes prevalence in the adult U.S. population. We advocate the inclusion of family history assessment in public health prevention and screening programs as an inexpensive and valuable source of genomic information and measure of diabetes risk. PMID- 15888231 TI - Childhood obesity -- what we can learn from existing data on societal trends, part 2. AB - The number of overweight and obese youth has increased in recent decades, yet few data assess how the lives of children have changed during the "obesity epidemic." Part 1 of this two-part study discussed trends in time use, studying at home, and media use. Part 2 focuses on transportation, physical education, and diet. Walking or biking for transportation can expend a large amount of energy, but active transportation is not a major source of physical activity for youth, averaging eight minutes a day in 2001, with little change over the past few decades. For adolescents, there was no clear trend in physical education during the past decade, but there are no data for after-school and day-care programs, which have become more important as children spend more time away from home. For younger children, time spent in organized sports and outdoor activities increased by 73 minutes per week between 1981 and 1997. Eating as a primary activity declined, suggesting a shift toward snacking or eating as a secondary activity. Statistically significant trends exist for carbohydrate intake, especially for chips/crackers/popcorn/pretzels (intake tripled from the mid-1970s to the mid 1990s) and soft drinks (intake doubled during the same period). Price and income data suggest possible economic reasons for these changes. The percentage of disposable income spent on food has declined continuously, and almost all of the decline has been represented by food consumed at home, yet today's disposable income buys more calories than it has in the past. Relative prices have encouraged shifts across food types. From a baseline of 100 during 1982-84, the price index for fresh fruit and vegetables increased to 258 by 2002 (far exceeding general inflation), whereas the price index for soft drinks increased only to 126 by 2002 (below general inflation). PMID- 15888233 TI - Capturing change in a community--university partnership: Si Se Puede! Project. AB - BACKGROUND: Community health interventions are increasingly employing partnerships combined with multilevel intervention models to achieve their objectives. Resources and methods for project evaluation are often limited to changes in population health status or health behaviors, while broader contextual questions that may illuminate mechanisms for change across ecological levels and project sustainability may not be addressed. CONTEXT: This paper describes a project to prevent and control diabetes in a Latino community and presents practical methods for addressing some challenges to evaluation, using data sources that often may be overlooked. METHODS: A case study method was used to examine approaches to capture data that can help explain changes across ecological levels. An ecological framework was used to organize sources of data. Data sources and findings are related to project timelines and goals. CONSEQUENCES: Although not a direct focus of the original research, substantial changes in community capacity were observed and measured over the course of the five-year project. Documentation on community change was found in routine project reports, logs, the news media, meeting minutes, and community documents. INTERPRETATION: A logical progression of community change across ecological levels became evident. A modest post hoc evaluation was feasible, using data routinely available from project and target community sources. Specific questions for future research on how community change occurs and how such changes may relate to population health and sustainability are suggested. PMID- 15888234 TI - Blood Pressure Sunday: introducing genomics to the community through family history. AB - BACKGROUND: Family history of a chronic disease, such as high blood pressure, is an important predictor of future disease. The integration of genomics information into public health activities offers the opportunity to help raise awareness among populations at high risk for high blood pressure. CONTEXT: The prevalence of high blood pressure in blacks at any age is about twice that of whites. Detroit is second among major U.S. cities in the percentage of residents who are black (81.6%). According to data from the Behavioral Risk Factor Surveillance System 1998-2002, the perceived health status of Detroit respondents was one of the worst in Michigan; 17.4% of Detroit respondents reported no health care coverage; 69.6% reported being obese or overweight; and 33.1% reported no physical activity. METHODS: The Michigan Department of Community Health and the University of Michigan's Center for Genomics and Public Health collaborated on a pilot program to develop a worksheet emphasizing the importance of personal family history of high blood pressure. The handout was distributed to individuals at primarily black, Detroit-area churches during an annual screening event for high blood pressure and stroke. CONSEQUENCES: Approximately 500 handouts were distributed; a collaborative effort was achieved; genomics information was integrated into an existing program; the ability to reach churches in a predominantly black community was demonstrated; consumers reported interest in the subject matter; and an appropriate literacy level for the handout was attained. INTERPRETATION: The strengths of this pilot program and suggested modifications may serve to guide others in genomics and/or chronic disease programs in future endeavors. PMID- 15888235 TI - Utah's Family High Risk Program: bridging the gap between genomics and public health. AB - BACKGROUND: Family history is a simple yet powerful genomic tool that can identify individuals and entire populations at risk for diseases such as heart disease, cancer, and diabetes. Despite its use for predicting disease, family history has traditionally been underused in the public health setting. CONTEXT: A program for identifying families at risk for a variety of chronic diseases was implemented in Utah. Utah has population characteristics that are unique among the United States. Although the land area is large, most residents live within a relatively small geographic area. The religion of 70% of the residents encourages the recording of detailed family histories, and many families have access to records dating back to the 1800s. METHODS: From 1983 through 1999, the Utah Department of Health, local health departments, school districts, the University of Utah, and the Baylor College of Medicine implemented and conducted the Family High Risk Program, which identified families at risk for chronic diseases using the Health Family Tree Questionnaire in Utah high schools. CONSEQUENCES: The collection of family history is a cost-effective method for identifying and intervening with high-risk populations. More than 80% of eligible families consented to fully participate in the program. A total of 80,611 usable trees were collected. Of the 151,188 Utah families who participated, 8546 families identified as high-risk for disease(s) were offered follow-up interventions. INTERPRETATION: The program was revolutionary in design and demonstrated that family history can bridge the gap between genetic advances and public health practice. PMID- 15888236 TI - Genomics and public health: development of Web-based training tools for increasing genomic awareness. AB - In 2001, the Centers for Disease Control and Prevention funded three Centers for Genomics and Public Health to develop training tools for increasing genomic awareness. Over the past three years, the centers, working together with the Centers for Disease Control and Prevention's Office of Genomics and Disease Prevention, have developed tools to increase awareness of the impact genomics will have on public health practice, to provide a foundation for understanding basic genomic advances, and to translate the relevance of that information to public health practitioners' own work. These training tools serve to communicate genomic advances and their potential for integration into public heath practice. This paper highlights two of these training tools: 1) Genomics for Public Health Practitioners: The Practical Application of Genomics in Public Health Practice, a Web-based introduction to genomics, and 2) Six Weeks to Genomic Awareness, an in depth training module on public health genomics. This paper focuses on the processes and collaborative efforts by which these live presentations were developed and delivered as Web-based training sessions. PMID- 15888237 TI - Prevention Research Centers: contributions to updating the public health workforce through training. AB - Because public health is a continually evolving field, it is essential to provide ample training opportunities for public health professionals. As a natural outgrowth of the Centers for Disease Control and Prevention's Prevention Research Centers Program, training courses of many types have been developed for public health practitioners working in the field. This article describes three of the Prevention Research Center training program offerings: Evidence-Based Public Health, Physical Activity and Public Health for Practitioners, and Social Marketing. These courses illustrate the commitment of the Prevention Research Centers Program to helping create a better trained public health workforce, thereby enhancing the likelihood of improving public health. PMID- 15888238 TI - The asthma consultative process: a collaborative approach to integrating genomics into public health practice. AB - Genomics research findings on asthma are reported with increasing frequency. As these findings are incorporated into existing knowledge of disease etiology and pathogenesis, the implications for public health practice need to be considered. In 2003, the University of Washington Center for Genomics and Public Health initiated a project to synthesize information about asthma genomics, to examine its relevance to public health research and practice, and to communicate findings to a public health practice audience. This goal was achieved through review of the scientific literature, formation of a working group, and consultations with professionals and community organizations. This paper describes the methods used to conduct these professional and community consultations, referred to as the asthma consultative process, and discusses the lessons learned from this activity. PMID- 15888239 TI - A note on "A catalog of biases in questionnaires". PMID- 15888241 TI - Expression of the high-affinity laminin receptor (67 kDa) in normal human skin and appendages. AB - The interaction of cells with extracellular matrix components plays a significant role in the regulation of cell biology. Laminin is a large glycoprotein involved in fundamental interactions between cells and the basement membrane. Several cell surface receptors are responsible for cell-matrix interactions. The 67 kDa high affinity laminin receptor, 67LR, is involved in the adhesion of normal cells to the laminin network and is also associated with the metastatic phenotype of some tumoral cells. We have investigated the expression of laminin and of the 67LR in normal human skin using immunoperoxidase staining. Twenty samples of skin were analyzed. Antibody against laminin reacted in a continuous linear band at the dermal-epidermal junction, as well as basement membranes of hair follicles, sebaceous and eccrine sweat glands, and dermal blood vessels. The epidermis and the follicular epithelium were negative for laminin. The 67LR seemed not to be expressed on the basal surface of basal keratinocytes. The major expression of this receptor may be detected in the upper half of the spinous layer and in the granular layer. The cells of the outer root sheath in hair follicle showed the same immunohistochemical pattern described for epidermis. In sebaceous glands and in eccrine sweat glands the secreting epithelium was positive. Endothelial cells of dermal blood vessels were routinely positive for 67LR. We observed that the expression of the 67LR in normal human skin is mostly located in epidermal areas in which the keratinizing process was particularly advanced. PMID- 15888242 TI - Interleukin 27 (IL-27): A novel pleiotropic cytokine involved in T cell differentiation and T cell response modulation. PMID- 15888243 TI - Mesenteric traction syndrome or gut in distress. AB - The mesenteric traction syndrome (MTS) remains, seventeen years after its first designation as a distinct pathologic entity, an aspect of surgical practice that has received limited attention. Apart from its symptomatology, there is mostly speculation about its exact causes and pathophysiologic mechanisms. Even though full-blown MTS is rather rare, the advent of laparoscopic surgery has indicated that there are different stages of stress associated with open abdominal surgery. Some evidence points to the involvement of mast cell derived vasoactive mediators and suggests possible pharmacologic management. PMID- 15888244 TI - The regulation of feeding: a cross talk between peripheral and central signalling. AB - Feeding and energy expenditures are modulated by the interplay of hormones and neurotransmitters in the central nervous system (CNS), where the hypothalamus plays a pivotal role in the transduction of peripheral afferents into satiety and feeding signals. Aminergic neurotransmitters such as dopamine (DA), norepinephrine (NE) and serotonin (5-hydroxytryptamine, 5-HT) are historically considered to play a key role, but a number of peptides are involved in finely tuning feeding regulation. This review summarizes the current understanding of the CNS mechanisms of orexigenic peptides, such as neuropeptide Y, orexins, and ghrelin, as well as anorectic peptides, such as leptin, neurotensin (NT), cocaine and amphetamine regulated transcript (CART) peptide, thyrotropin-releasing hormone (TRH), corticotropin-releasing hormone (CRH), urocortin, amylin. PMID- 15888245 TI - Morphological and cytofluorimetric analysis of adult mesenchymal stem cells expanded ex vivo from periodontal ligament. AB - Many adult tissues contain a population of stem cells that have the ability of regeneration after trauma, disease or aging. Recently, there has been great interest in mesenchymal stem cells and their roles in maintaining the physiological structure of tissues, and their studies have been considered very important and intriguing, after having shown that this cell population can be expanded ex vivo to regenerate tissues not only of the mesenchymal lineage, such as intervertebral disc cartilage, bone, tooth-associated tissue, cardiomyocytes, but also to differentiate into cells derived from other embryonic layers, including neurons. Currently, different efforts have been focused on the identification of odontogenic progenitors from oral tissues. In this study we isolated and characterized a population of homogeneous human mesenchymal stem cells proliferating in culture with an attached well-spread morphology derived from periodontal ligament, a tissue of ectomesenchymal origin, with the ability to form a specialized joint between alveolar bone and tooth. The adherent cells were harvested and expanded ex vivo under specific conditions and analysed by FACScan flow cytometer and morphological analysis was carried out by light, scanning and transmission electron microscopy. Our results displayed highly evident cells with a fibroblast-like morphology and a secretory apparatus, probably indicating that the enhanced function of the secretory apparatus of the mesenchymal stem cells may be associated with the secretion of molecules that are required to survive and proliferate. Moreover, the presence in periodontal ligament of CD90, CD29, CD44,CD166, CD 105, CD13 positive cells, antigens that are also identified as stromal precursors of the bone marrow, indicate that the periodontal ligament may turn out to be a new efficient source of the cells with intrinsic capacity to self-renewal, high ability to proliferate and differentiate, that can be utilized for a new approach to regenerative medicine and tissue engineering. PMID- 15888246 TI - Xaliproden (SR57746A) induces 5-HT1A receptor-mediated MAP kinase activation in PC12 cells. AB - Neurotrophic growth factors are involved in cell survival. However, natural growth factors have a very limited therapeutic use because of their short half life. In the present study, we investigated the mechanism of action of a non peptidic neurotrophic drug, Xaliproden, a potential molecule for the treatment of motoneuron diseases, since the transduction pathways of this synthetic 5-HT1A agonist are very poorly understood. Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the ERK1 and ERK2 isoforms of MAP kinase, which then rapidly decrease to the basal level. We demonstrate that isoforms of the SHC adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced MAP kinases activation. The inhibitor of Ras farnesylation, FPT-1, and the protein kinase C inhibitors, GF 109203X and chelerythrine, inhibited the Xaliproden-induced MAP kinase activation, suggesting p21Ras and PKC involvement. Moreover, the observations that the 5-HT1A antagonist, pindobind, and pertussis toxin abolished the Xaliproden-induced ERK stimulation suggested that Xaliproden activates the MAP kinase pathways by stimulating the G protein-coupled receptor, 5-HT1A. These results demonstrate clearly that the non-peptidic compound, Xaliproden, exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins. These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by MAP kinase pathway by a pertussis toxin-sensitive mechanism involving 5-HT1A receptors, p21 Ras and MEK-1 and by PKC and Akt pathways. PMID- 15888247 TI - In vitro effects of an immunostimulating bacterial lysate on human lymphocyte function. AB - MLBL is an oral immunostimulating vaccine consisting of bacterial standardized lysates obtained by mechanical lysis of different strains of Gram-positive and Gram-negative bacteria that can cause acute and chronic infections of the respiratory tract. Previous studies suggested a stimulating effect of MLBL both on humoral and cellular immune responses. In the present study, the in vitro effects of MLBL on human lymphocyte effector functions and its mechanisms of action were evaluated. The results show that the most remarkable effects of MLBL on the immune system are: i) activation of the IL-2 receptor (IL-2Ralpha) on different lymphocyte subsets (B, CD4+ T and CD8+ T cells) involved both in humoral and cellular immune responses; ii) induction of cytokine synthesis (IL-2, IL-10, IL-12, IFNgamma) in the immune competent cells that induce and regulate immune responses; iii) generation of CD4+ and CD8+ effector T cells. Overall, these results suggest that the therapeutic effect of MLBL on acute and recurrent infections of the respiratory tract is related to its ability to activate the responses of different subsets of immune competent cells both for humoral and cellular immunity. Moreover, these effects can be induced either by direct immune cell activation or through the generation and activation of immune effector cells. PMID- 15888248 TI - P2Y1 and cysteinyl leukotriene receptors mediate purine and cysteinyl leukotriene co-release in primary cultures of rat microglia. AB - Inflammation is widely recognized as contributing to the pathology of acute and chronic neurodegenerative conditions. Microglial cells are pathologic sensors in the brain and activated microglia have been viewed as detrimental. Leukotriene, including cysteinyl leukotrienes (CysLTs) are suggested to be involved in brain inflammation and neurological diseases and ATP, by its receptors is a candidate for microglia activation. A23187 (10 microM) stimulated microglia to co-release CysLTs and [3H] adenine based purines ([3H] ABPs), mainly ATP. The biosynthetic production of CysLTs was abolished by 10 microM MK-886, an inhibitor of 5 lipoxygenase-activating protein activity. RT-PCR analysis showed that microglia expressed both CysLT1 / CysLT2 receptors, P2Y1ATP receptors and several members of the ATP binding cassette (ABC) transporters including MRP1, MRP4 and Pgp. The increase in [Ca2+]i elicited by LTD4 (0.1 microM) and 2MeSATP (100 microM), agonists for CysLT- and P2Y1-receptors, was abolished by the respective antagonists, BAYu9773 (0.5 microM) and suramin (50 microM). The stimulation of both receptor subtypes, induced a concomitant increase in the release of both [3H] ABPs and CysLTs that was blocked by the antagonists and significantly reduced by a cocktail of ABC transporter inhibitors, BAPTA/AM (intracellular Ca2+ chelator) and staurosporine (0.1 microM, PKC blocker). P2Y antagonist was unable to antagonise the effects of LTD4 and BAYu9773 did not reduce the effects of 2MeSATP. These data suggest that: i) the efflux of purines and cysteinyl leukotrienes is specifically and independently controlled by the two receptor types, ii) calcium, PKC and the ABC transporter system can reasonably be considered common mechanisms underlying the release of ABPs and CysLTs from microglia. The blockade of P2Y1 or CysLT1/CysLT2 receptors by specific antagonists that abolished the raise in [Ca2+]i and drastically reduced the concomitant efflux of both compounds, as well as the effects of BAPTA and staurosporine support this hypothesis. In conclusion, the data of the present study suggest a cross talk between the purine and leukotriene systems in a possible autocrine/paracrine control of the microglia-mediated initiation and progression of an inflammatory response. PMID- 15888249 TI - HLA-E up-regulation induced by HIV infection may directly contribute to CD94 mediated impairment of NK cells. AB - Alterations in NK cell numbers and function have been repeatedly shown during HIV infection. In this study, NK cell number and MHC class I expression on CD4+ T cells were studied in HIV patients at different stages of disease progression. An increased expression of HLA-E was seen on CD4+ T cells. In parallel, a reduced number of CD94+ NK cells was observed in advanced disease stages. Moreover, a decline in CD94 expression on NK cells was observed at the HIV replication peak in patients undergoing antiretroviral treatment interruption, suggesting a role of viral replication on NK cells alterations. In vitro HIV infection induced a rapid down-regulation of HLA-A,B,C expression, paralleled by an increased expression of HLA-E surface molecules, the formal ligands of CD94 NK receptors. HIV-infected HLA-E expressing cells were able to inhibit NK cell cytotoxicity through HLA-E expression, since cytotoxicity was restored by antibody masking experiments. These data indicate that the CD94/HLA-E interaction may contribute to NK cell dysfunction in HIV infection, suggesting a role of HIV replication in this process. PMID- 15888250 TI - A pilot study of feasibility of ultra-rush (20-25 minutes) sublingual-swallow immunotherapy in 679 patients (699 sessions) with allergic rhinitis and/or asthma. AB - Few studies have evaluated the occurrence of immediate adverse reactions in allergic patients after an ultra-rush regimen of different commercial allergen extracts for sublingual immunotherapy (SLIT) Methods: 679 patients took part in trials of specific ultra-rush SLIT for the treatment of IgE-mediated rhinitis and/or IgE-mediated asthma. 14 patients received two different sublingual allergen vaccines during two distinct SLIT sessions. On the whole, 699 SLIT sessions were performed. The build up ultrarush phase involved the administration every five minutes of increasing doses of either different allergen extracts. The cumulative allergen extract solution after half an hour was several times the dose administered at the start of subcutaneous immunotherapy (range 4.7-525 microg of major allergens). All patients tolerated the treatment very well. 122 (17.96%) had mild local symptoms (pruritus of the buccal cavity) that spontaneously disappeared with increasing dose. Two patients allergic to Parietaria had urticaria about three hours after the last sublingual Parietaria extract intake. A subject allergic to Artemisia vulgaris pollen had urticaria and rhinitis two hours later than the last dose of vaccine. As reported in our previous study, no immediate severe adverse reactions were observed after that rapidly increasing doses of allergen extract were administered in a very short period to a large number of patients, showing the excellent safety profile of ultra-rush SLIT. PMID- 15888251 TI - Serum evaluation of angiogenic cytokine basic fibroblast growth factor, hepatocyte growth factor and TNF-alpha in patients with myelodysplastic syndromes: correlation with bone marrow microvascular density. AB - Recent studies have documented that angiogenesis plays a significant role in haematological malignancies, including mylodysplastic syndromes (MDS). Basic fibroblast growth factor (b-FGF), Hepatocyte growth factor (HGF) and Tumor necrosis factor-alpha (TNF-alpha) are multifunctional cytokines that potently stimulate angiogenesis. The aim of the present study was to evaluate the microvascular density (MVD) and the serum levels of these angiogenic factors in patients with myelodysplastic syndromes (MDS). In 61 patients with MDS, MVD was measured in bone marrow biopsies and b-FGF, HGF and TNF-alpha were determined in the serum of the same patients by enzyme-linked immunosorbent assay (ELISA). Serum levels of b-FGF, HGF and TNF-alpha as well as MVD in the bone marrow were increased in MDS patients compared to healthy controls (p<0.0001). Levels of b FGF, HGF and TNF-alpha were also significantly higher in high-risk for leukemic transformation MDS than in low-risk (p<0.0001). Significant differences were also found regarding MVD in high and low risk patients (p<0.001). Both b-FGF and HGF levels were significant predictors of survival (p<0.0005, log-rank test). The present study showed that serum levels of b-FGF, HGF and TNF-alpha are significantly increased and dependent on the severity of MDS suggesting that the determination of these parameters may offer considerable information regarding disease progression and prognosis. PMID- 15888252 TI - Amelioration of neutrophil membrane function underlies granulocyte-colony stimulating factor action in glycogen storage disease 1b. AB - Glycogen storage disease (GSD) 1b is a metabolic disorder characterized by a deficiency of glucose 6-phosphate transporter and neutrophil alterations, which are reduced in number and functionally impaired. The present study aimed at investigating neutrophil dysfunction correlating submembrane and cytoskeletal changes at different ages with or without granulocyte-colony stimulating factor (G-CSF) treatment. GSD1b neutrophils showed reduced expression and diffused localization of focal adhesion kinase (FAK) and actin. No abnormalities were observed in GSD1a patient neutrophils. Gelsolin was also slightly reduced in neutrophils of GSD1b patients. When patients were treated for at least 3 months with G-CSF, the neutrophil number and the expression of FAK and actin were significantly increased. Granulocyte colony-stimulating factor treatment was similarly effective when performed in 1 year old patients. FAK auto- and IL-8 mediated phosphorylations were already affected as early as 1 year of age. G-CSF treatment also improved this alteration. Our data suggest that neutrophil dysfunction in GSD1b patients might be related to functional impairment and disorganization of proteins of the sub-membrane apparatus, and that G-CSF treatment counteracts neutropenia and prevents the progressive alterations of neutrophil sub-membrane proteins. PMID- 15888253 TI - BKV infection and hemorrhagic cystitis after allogeneic bone marrow transplant. AB - Hemorrhagic cystitis (HC) is a well-known complication after allogeneic bone marrow transplant (BMT) and can be related to adenovirus or human polyomavirus BK (BKV) infections. In this study a group of 20 patients after allogeneic BMT has been examined. BMT urine samples were analysed for the presence of Adenovirus and BKV DNAby means of polymerase chain reaction (PCR). 5/20 BMT patients developed HC after BMT. The presence of BKV DNA in urine samples was evident in 3/15 patients without HC and in 5/5 patients with HC. In 2/5 HC-patients the BKV DNA was not found after therapy with Cidofovir and Ribavirin. The search for adenovirus DNA in all samples was negative. The analysis of BKV non-coding control region (NCCR) isolated from urine samples revealed a structure very similar to the archetype in all samples. The RFLP (Restriction Fragment Length Polymorphism assay) showed the presence of BKV subtypes I and IV, with the prevalence of subtype I (4/5). This study supports the hypothesis that HC is mainly related to BKV rather than to adenovirus infection in BMT patients. Moreover, since BKV subtype I was predominant, it is reasonable to hypothesize that a specific BKV subtype could be associated with the development of HC. PMID- 15888254 TI - Apoptotic death of Listeria monocytogenes-infected human macrophages induced by lactoferricin B, a bovine lactoferrin-derived peptide. AB - Listeria monocytogenes, an intracellular facultative food-borne pathogen, was reported to induce apoptosis in vitro and in vivo in a variety of cell types with the exception of murine macrophages. These cells represent the predominant compartment of bacterial multiplication and die as a result of necrosis. In this study we showed that human non-activated and IFN-gamma-activated macrophagic-like (THP-1) cells infected with L. monocytogenes, mainly die by necrosis rather than by an apoptotic process. Two natural products derived from bovine milk, lactoferrin and its derivative peptide lactoferricin B, are capable of regulating the fate of infected human macrophages. Bovine lactoferrin treatment of macrophages protects them from L. monocytogenes-induced death whereas lactoferricin B, its derivative peptide, determines a shifting of the equilibrium from necrosis to apoptosis. PMID- 15888255 TI - CD40-dependent activation of phosphatidylinositol 3-kinase/AKT pathway inhibits apoptosis of human cultured mesangial cells induced by oxidized LDL. AB - Deposition of atherogenic lipoproteins is associated with various glomerular diseases. In particular, oxidized LDL (oxLDL) may affect mesangial cells and favour the development of glomerulosclerosis. The aim of the present study was to investigate on cultured human mesangial cells (HMC) whether oxLDL induces apoptosis by a mechanism dependent on the inhibition of Akt survival pathway, and whether the engagement of mesangial CD40 by its ligand CD154 inhibits the apoptotic effect of oxLDL. Tunel assays demonstrated that incubation of HMC for 24 h with oxLDL, but not with unmodified LDL, induced a dose-dependent increase in apoptosis of HMC associated with a decrease in Akt phosphorylation. Enzymatic kinase assay showed that also the Akt activity was reduced in a dose-dependent manner by treatment with oxLDL. Stimulation of mesangial CD40 with sCD154 rescued HMC from oxLDL-dependent apoptosis, while two unrelated pharmacological inhibitors of PI3K LY294002 and wortmannin abrogated this anti-apoptotic effect, suggesting an involvement of the PI3K/Akt pathway. Moreover CD40 stimulation maintained an elevated phosphorylation of Akt and preserved its enzymatic activity in the presence of oxLDL. Indeed, CD154 induced a rapid enhancement in Akt enzymatic activity, that was temporarily correlated with the association of CD40 with TRAF3, TRAF6, c-Cbl and the p85 subunit of PI3K. In conclusion, these results suggest that CD40 stimulation protects HMC from toxic effects of oxLDL by promoting PI3K/Akt-dependent cell survival. PMID- 15888256 TI - IL-10 enhances CCL2 release and chemotaxis induced by CCL16 in human monocytes. AB - CCL16 is a CC chemokine originally identified as a liver-expressed chemokine. Its expression has been detected in activated monocytes where it is up-regulated by stimulation with IL-10. This is in contrast with IL-10's inhibition of the expression of most chemokines. CCL16 is chemotactic for monocytes, lymphocyte and dendritic cells. We investigated whether CCL16 displays biological activities other than chemotaxis and whether IL-10 affects monocyte response to CCL16. We show that CCL16 induces the expression of CCL2 at the mRNA and protein level, but does not affect that of CCL5, CCL18 and proinflammatory cytokines. This effect was prevented by treatment with pertussis toxin and may thus be mediated by G protein-coupled receptors. IL-10 markedly increased CCL2 production induced by CCL16, but suppressed that of CXCL8. It also enhanced the chemotactic response to CCL16. Addition of antibodies blocking CCR1, but not CCR8, prevented this enhanced chemotactic response and suggested that CCR1 is primarily involved. We propose that IL-10 modulates the effects of CCL16 on monocytes by increasing their CCR1-dependent response. The coordinated secretion of CCL16 and IL-10 may thus enhance monocyte infiltration. PMID- 15888257 TI - A limited autoimmunity to p185neu elicited by DNA and allogeneic cell vaccine hampers the progression of preneoplastic lesions in HER-2/NEU transgenic mice. AB - Prevention of the progression of precancerous lesions by vaccines is virtually uncharted territory. Their potential, however, is being assessed in transgenic mice which develop autochthonous tumors with defined stages of progression. In this paper we show that the DNA micro-array technology significantly helps assessment of the preventive efficacy of a combined DNA and cell vaccine. All female rat Her-2/neu transgenic BALB/c (BALB-neuT) mice develop an invasive carcinoma in each of their mammary glands within 25 weeks of age. This is elicited by the activated transforming rat Her-2/neu oncogene embedded in their genome. We have previously shown that vaccination of mice bearing multiple in situ carcinomas with DNA plasmids which code for the extracellular and transmembrane domain of rat p185neu, the product of the rat Her-2/neu oncogene, followed by a boost with rat p185neu+ allogeneic cells engineered to secrete interferon-gamma, keeps 48% of mice tumor free until week 32. We have now extended our follow-up until mice reach one year of age and show that protection vanishes as time progresses. This observation suggests that the accuracy of the results studying immunotherapy against life-threatening tumors is a function of the length of the follow-up. The application of microarrays, and the concordance of morphologic and gene expression data led us to identify antibody as the main mechanism induced by vaccination. Protection is associated with a break of tolerance and a limited autoimmunity against the endogenous mouse p185neu. PMID- 15888258 TI - Excessively high magnetic resonance signal in preterm infants and neuropsychobehavioural follow-up at 2 years. AB - The diffuse excessive high-signal intensity (DEHSI) findings in the T2 weighted scans of white matter (WM), besides the corresponding low signal in the T1 weighted images, are usually more evident around the periventricular regions. It is not clear whether the DEHSI should be considered as a diffuse WM injury rather than a sign of delayed maturation of the WM. Eighty nine preterm infants at the full-term equivalent age (FEA) were studied using conventional Magnetic Resonance (MR) imaging of the brain. Based on the MR findings, the infants studied were divided into three groups: the control group presenting normal WM, the DEHSI group and the group with other WM lesions. Ten newborns were not included in the statistical analysis because they presented evidence of precedent germinal matrix hemorrhage (GMH-IVH) which cannot be considered as WM lesions. Seventy nine infants were enrolled in a program of neuropsychobehavioural study follow-up until 24 months of age. Each infant was evaluated for those variables which mostly affect the occurrence of neuropsychomotor disability. In the DEHSI infant group, significantly lower mean pH and mean base excess (BE) values were found in comparison to controls, while the mean birth weight (BW) was significantly higher. No significant difference was observed between the mean 1st minute Apgar Score, mean birth gestational age (GA) and assisted ventilation mean duration of controls and DEHSI groups. Finally, no significant difference between the parameters studied was found by comparing the WM lesion infants group to the DEHSI infants one. Our observations, together with follow-up studies, even up to school age, confirm that DEHSI has a clinical significance and cannot be considered as a simple indicator of delayed WM maturation. PMID- 15888259 TI - Effects of interferon beta, cyclophosphamide and azathioprine on cytokine profile in patients with multiple sclerosis. AB - We assessed the in vitro effects of interferon beta-1b (IFNbeta-1b), cyclophosphamide (CY), and azathioprine (AZA) alone and of the combination of IFNbeta-1b with CY or AZA on the production of Th1 and Th2 cytokines in 10 patients with multiple sclerosis. Cytokine levels were determined at baseline and after stimulation with IFNbeta-1b, CY, and AZA alone or with the combination of IFNbeta-1b with CY or AZA. The combination of IFNbeta-1b with CY resulted in a statistically significant decrease in the production of interleukin-2 (IL-2) (P=0.003) and tumor necrosis factor alpha (TNF-alpha) (P=0.03). An additive effect on the production of interferon gamma (IFN-gamma) (P=0.2) and interleukin 10 (IL-10) (P=0.6), and a positive interaction on the production of interleukin-4 (IL-4) (P=0.08) were observed although the findings were not statistically significant. The combination of IFNbeta-1b with AZA resulted in a significant negative effect on the production of IL-2 (P=0.006), whereas TNF-alpha (P=0.02), IFN-gamma (P=0.03), IL-4 (P=0.2), and IL-10 (P=0.3) were not statistically impacted. Our data show that CY was able to improve the effects of IFNbeta-1b on the ratio of Th1/Th2 cytokines. PMID- 15888261 TI - [Informed consent: more than research authorization]. PMID- 15888260 TI - Acute polyradiculoneuropathy occurring after hymenoptera stings: a clinical case study. AB - Hymenoptera stings may be responsible for both local and systemic reactions; these can be immediate or delayed, depending on the time between the sting and the development of signs or symptoms. Delayed clinical reactions have been reported, although unusual, due to serum sickness and/or affecting organs or systems generally not involved in the immediate reaction, such as heart, kidneys, central and peripheral nervous systems. This paper describes the clinical and immunological findings in a 51-year-old subject, who, after two stings of paper wasps, the second one after the third venom immunotherapy (VIT) injection, presented immediate large local and systemic allergic reactions which quickly improved after e.v. methylprednisolone administration. About 40 hours later, he developed acute polyradiculoneuropathy with muscle weakness, paresthesia, difficulties in standing up and walking. Skin tests and specific IgE determination showed allergy to paper wasp. The activation, by wasp venom, of peripheral blood mononuclear cells in primary culture, evaluated by tritiated thymidine incorporation proliferation assay, showed an important hypersensitivity to wasp venom. Therefore our results suggest the hypothesis that the polyradiculoneuritis causative etiopathogenetic mechanism might be a delayed immunological response to wasp antigens followed by an allergy-triggered autoimmune reaction, as previously suggested by other authors; they found lymphocytic infiltrates in demyelinization areas and at perivascular levels, by histologic examination of autoptical and bioptical material of patients with nervous system lesions after hymenoptera stings. PMID- 15888262 TI - [Thyroidectomy associated with laryngectomy in laryngeal cancer treatment. Is it routinely necessary?]. AB - INTRODUCTION: In patients with laryngeal invasive epidermoid carcinoma who are candidates for total laryngectomy, it is recommended to resect en bloc at least half of the thyroid gland on the same side as the laryngeal tumor with the objective of decreasing local recurrence associated with thyroid infiltration. Nevertheless, in the histopathologic analysis of the specimen, a minority of thyroid glands show tumor infiltration. The fact that in these patients even partial thyroid resection is associated with hypothyroidism increased by postoperative radiotherapy is well known. The study was undertaken to determine the frequency of thyroid gland invasion in patients who have undergone total laryngectomy due to laryngeal cancer and its associated factors. MATERIAL AND METHODS: We evaluated the histological results of glands resected en bloc with laryngectomy in patients who underwent total laryngectomy due to laryngeal cancer; the clinical stage was compared to the laryngeal subsite tumor origin. The glandular histological condition was compared to the local recurrence presence (peristomal). RESULTS: Ninety two patients were included, 11 (12%) showed glandular infiltration due to epidermoid carcinoma, all with tumors clinically typified as T3 and T4, 8/11 were transglottic and only 3 (27%) showed subglottic invasion. During follow-up (5-year mean) 17/92 showed peristomal recurrence (18%), only 3 (3%) showed thyroid invasion. Among patients with glandular invasion the peristoma recurrence rate was 27% (3/11). DISCUSSION: The best laryngeal cancer thyroid tumor invasion predictor is the evidence of extralaryngeal extension. Thyroid resection en bloc should not be routinely advised due to the low frequency of glandular infiltration. PMID- 15888263 TI - Delphian lymph node in glottic carcinoma subjected to supracricoid partial laryngectomy with cricohyoidoepiglottopexy. AB - INTRODUCTION: Metastases to Delphian lymph node is rarely present in laryngeal malignancy. This report describes its frequency in patients with glottic cancer undergoing supracricoid partial laryngectomy (SCPL) with cricohyoidoepiglottopexy (CHEP). MATERIAL AND METHODS: Fourteen patients (13 male and 1 female) with a mean age of 58 years underwent a SCPL with CHEP and functional bilateral neck dissection (levels II-V) searching for the Delphian lymph node. Four patients were in stage I, five in stage II, four in stage III, and one in stage IV. Surgical margins and Delphian lymph nodes were searched for in each partial laryngectomy sample. RESULTS: The mean follow-up was 9 months. The right-side dissection yielded an average of 18 lymph nodes and the left-side dissection yielded an average of 22 lymph nodes, with no metastatic disease on the ultimate examination. Only one patient (7%) revealed a carcinoma-positive Delphian lymph node, and 6 months later a metastatic lymph node was found on the same side as the primary tumor. The patient underwent standard radiation therapy (66 Gy) targeted to larynx and lymph node areas, as well as to the supraclavicular region (20 Gy). DISCUSSION: Bilateral functional dissection is indicated in the presence of Delphian lymph node metastatic spread. Post-operative radiation therapy may occasionally be used as an adjuvant treatment in cases with positive Delphian lymph node in spite of a negative functional dissection in partial laryngectomies due to other possible node spread routes. Bilateral functional dissection should be chosen based on tumor site and size. PMID- 15888264 TI - [Factors related to the olive size in patients with hypertrophic pyloric stenosis]. AB - INTRODUCTION: The pyloric "olive" (PO) is the result of the anomalous growth of the pyloric muscle among patients with pyloric stenosis (PS). It frequently is unexpectedly large, or some other times surprisingly small, and those variations in size have been difficult to explain. MATERIALS AND METHODS: We measured the PO in 145 consecutive patients with PS during the operation, and then we classified them as small if their length was less than 20 mm, medium if 20 to 30 mm, or large if more than 30 mm; several variables were analyzed by mean of the chi square or Spearman rho tests. RESULTS: Six cases were excluded due to an unclear total length record of the PO. The PO size was classified as small in 19 (13.7%), medium in 71 (51%) and large in 49 (35.3%). The PO size did not associate with gender, way of birth, the presence of jaundice, constipation or any specific blood group or Rh factor, and it did not correlate with birth weight or month and gestational order either. Medium and large PO were more frequently palpated than smaller (94-100% vs. 83%, p = 0.009); PO size correlated with the duration of the history of vomiting (CQ 0.267, p = 0.002), child's age (CQ 0.243, p = 0.005) and weight at operation (CQ 0.190, p = 0.048). A daily weight loss surpassing 5 g was more commonly found among small PO (p = 0.038). CONCLUSIONS: In more than a third of the PS patients, PO is unexpectedly large, and in one of every seven it is surprisingly small. The bigger PO size associates with a longer disease, and with older and heavier patients, which probably is explained by a slighter clinical course. A small PO is more difficult to palpate during clinical evaluation. PMID- 15888265 TI - [Safety of laparoscopic cholecystectomy in complicated vesicular disease]. AB - OBJECTIVE: We assessed the experience and safety of cholecystectomy through laparoscopic approach in patients with complicated biliary gallbladder disease who were attended at the Hospital Espanol, Veracruz, Mexico, during a 10-year period. MATERIAL AND METHODS: A prospective, longitudinal, comparative study with a control group was designed. We studied a group of patients with complicated cholelithiasis disease (Group I) and compared them with patients without complicated diseases. We analyzed the following variables: age, gender, risk factors, associated trans-operative pathology and accidents, surgical time, rate of conversion to open procedure, length of hospital stay, complications and evolution. RESULTS: During the 10-year period, 733 cholecystectomies were performed, 245 (33.42%) to treat complicated cholelithiasis and 488 (66.58%) uncomplicated. There were no differences regarding gender, age, risk factors, hospital stay, complications, morbidity and mortality, and iatrogenic lesions of the biliary tree, postoperative morbidity and mortality. In all patients of Group I (245), we found complications of cholelithiasis (acute cholecystitis, choledochal gallstones, vesicular adherences, and cholecystocolonic fistula). Surgical time was longer in Group I and surgical accidents made the surgical procedure more difficult. CONCLUSIONS: The overall results established that laparoscopic cholecystectomy in our institution is a safe procedure in patients with complicated gallbladder disease and can be performed by experienced surgeons. PMID- 15888266 TI - [Epidemiology of xanthogranulomatous cholecystitis]. AB - OBJECTIVE: In order to study patients with a diagnosis of xanthogranulomatous cholecystitis (XGC), we analyzed their demographics, epidemiology and clinical data. MATERIAL AND METHODS: We analyzed the clinical records of XGC during a period of 6 years, obtaining demographic, epidemiologic and clinical data. RESULTS: Of a total of 1425 cholecystectomies performed between January 1991 and December 1996, we found 35 cases of XGC (2.4%). Twenty six (74%) were women (median age: 44 years), 60% were from a low socioeconomic group, 34% has a history of alcoholism and smoking, and 25 patients (71%) had a blood type of O positive. Thirteen patients (37%) presented obstructive jaundice, 11 had dilatation of the choledocus and were treated with ERCP. Of the 35 cholecystectomies, 15 were urgent and 20 elective. Eight were operated laparoscopically and two were converted because of firm adhesions. We had 5 transoperative complications. DISCUSSION: Pre-operative XGC diagnosis is difficult, often mistaken for gall bladder cancer. The incidence in our study (2.4%) is higher than reports in industrialized countries (0.7-1.8%), with a female predominance. The most frequent clinical presentation is that of chronic cholecystitis, but we found a high percentage of patients with obstructive jaundice. We had 0% mortality and 26% morbidity, and no association was found between XGC and gallbladder cancer. PMID- 15888267 TI - [Cost/benefit of preoperative examinations for routine elective surgery]. AB - INTRODUCTION: Preoperative exams are a common practice for surgeons; however, their use is not based on a scientific or medical basis. In the best of scenarios, it is an institutional policy. MATERIAL AND METHODS: We wanted to determine the utility and cost-effectiveness ratio of of preoperative routine tests (PRT) and we analyzed retrospectively the frequency of complications, hospital stay, and cost-effectiveness of PRT, in subjects with no concomitant disease, who were having elective surgery performed by the general surgery service. We included 141 male and female subjects, ages from 18 to 40 years, during 2002. RESULTS: There was no increase in complication frequency in patients with PRT abnormalities (6.2% versus 5.1%, OR 0.82, p = 0.78), or hospital stay time (2.37 versus 2.76, p = 0.55). PRT were repeated in 19.1%, expiration being the most frequent reason (46%). Urinalysis was the most frequent abnormal test (21.4%). We found only five clinically relevant abnormalities (3.5%). Only three patients required therapeutic intervention prior to surgery. The detection cost for PRT abnormalities requiring preoperative medical intervention was 22,732 Mexican pesos (approximately USD 2,022). CONCLUSIONS: The use of PRT in young clinically healthy subjects is an expensive and inefficient practice. PMID- 15888268 TI - [Bursting pressure in normal and ischemic colonic anastomoses in rats; using biological and synthetic adhesives]. AB - OBJECTIVE: We wanted to determine bursting pressure in normal and ischemic colonic anastomoses in rats as well as the frequency and type of complications with the use of biological and synthetic adhesives. MATERIAL AND METHODS: We designed an experimental study using 80 Sprague-Dawley rats, weighing between 200 and 250 g, divided into four groups: one control group and three study groups. Anastomoses were realized using one layer of interrupted 6-0 polypropylene sutures. Each group was divided into two categories: normal colon (n = 10) and ischemic colon, induced by dividing mesenteric blood vessels (n = 10). Group 2 used octyl-cyanoacrylate, group 3 used N-2-butyl-cyanoacrylate, and group 4 used 40 mg/ml fibrinogen with 1000 u/ml of thrombin. Bursting pressure was measured with a manometer creating pressure in the anastomotic bowel using an infusion pump at 1 ml/min of NaCl 0.9%. Statistical analysis was performed with Student's t test, one-way ANOVA, chi square test or Fisher's exact test. RESULTS: Group 1: (control) normal 127.8 +/- 16.21 versus 109 +/- 17 with ischemia (p < 0.05); group 2: normal 145.5 +/- 89 versus ischemic colon 97.6 +/- 40 (p = 0.136); group 3: normal 145.7 +/- 34 versus 130.8 +/- 15.33 with ischemia (p = 0.22); group 4: normal 239 +/- 26.4 versus 196.5 +/- 14.3 with ischemia (0.000). Bursting pressure was statistically significant in group 4, showing greater pressures (p < 0.001). Bursting segment was shown to be higher outside the anastomoses. Complications such as adhesions and intestinal obstruction were seen more frequently in both cyanoacrylate groups. CONCLUSIONS: An increased bursting pressure was shown in the fibrinogen groups, having a greater tensile strength of the anastomoses. Pressures similar when anastomoses were treated with any of the other two synthetic adhesives. PMID- 15888269 TI - [Intussusception in adults]. AB - Intestinal intussusception in adults is a very rare entity whose etiology differs greatly from its pediatric counterparts. Three adult patients with intestinal intussusception presented with intestinal obstruction and required surgery for its resolution. In each case a pathologic cause was found. They had intestinal polyps, two benign (lipoma and hamartomatous polyps) and one malignant (metastasis). Intestinal intussusception in adults usually has a pathologic cause. About half have a malignant etiology. Intestinal resection without reduction of the intussusception is the preferred surgical procedure. PMID- 15888270 TI - [Abdominal actinomycosis: report of three cases]. AB - We present three cases of abdominal actinomycosis in females, one presenting with an abdominal mass and the two others underwent emergency surgery because of acute abdomen with a diagnosis of complicated acute appendicitis. The first patient (age 36 years) presented with an abdominal mass in the left lower quadrant arising from the colon as observed by abdominal computed tomography (CT). The patient was brought to the operating room and tumoral resection was done. The second and third patients (37 years and 39 years, respectively) were brought to the emergency room because of acute abdominal pain with leucocytosis. Exploratory laparotomy was performed, finding in the second patient a bilateral ovarian abscess and uterine perforation. Hysterectomy and salpingo-oophorectomy were done. In the third patient, the findings were a sigmoid mass and a bilateral tubo ovarian abscess and these organs were resected. Samples were sent for pathologic analysis. Microscopic analysis of the specimens sent revealed the presence of "sulfur granules," and a diagnosis of actinomycosis was made. Abdominal actinomycosis is a rare disease and preoperative diagnosis is uncommon. It is necessary to complete the full course of antibiotic therapy in order to completely eradicate the disease. PMID- 15888271 TI - [Breast cancer associated with pregnancy]. AB - Invasive breast carcinoma diagnosed during pregnancy and even one year after delivery is known as breast cancer associated with pregnancy. There are many controversies surrounding this entity concerning diagnosis and therapeutics as well as to the behavior regarding the fetus. There is broad information in the existing literature. We can find authors who do not change the diagnostic therapeutic approach to the neoplasia and those who even advise pregnancy interruption and consider greater aggressiveness in treatment, in comparison to the same cancer in a non-pregnant woman. The prognosis of breast cancer associated with pregnancy has also been a controversial issue. Historically, it has been considered to have the worst evolution. Nevertheless, evidence recently has shown that the known prognosis factors in breast cancer (stage, nodular status, receptors, etc.) are those associated with the prognosis and not the pregnancy itself. The objective of this article is to analyze, based on the existing evidence in the literature, which is the best diagnostic-therapeutic route for breast cancer associated with pregnancy, which procedures must be followed with the mother and the fetus and which is the ideal therapeutic sequence, analyzing at the same time the possibilities of axillary preservation by means of lymphatic mapping and sentinel node biopsy in these patients. PMID- 15888272 TI - [Preventive vaccines and immunotherapy clinical trials against cervical cancer]. AB - Cervical cancer (CC) is a public health problem among women worldwide, especially in emerging nations. To improve CC control, new adjuvant therapeutic strategies are required. Advances in immunology, genomics and proteomics have accelerated our understanding of the genetic and cellular basis of many cancer types. CC is a member of virus-related neoplasms and its initiation and promotion is associated with persistent infection of oncogenic human papillomavirus. During viral infection and associated-transforming developing lesions, the HPVs co-express non structural and structural proteins. These early or late proteins are the antigenic target of the immune response. The intervention to stimulate the humoral or cellular immune anti-HPV response is the objective of the immunoprevention and immunotherapy against CC. Recently in a controlled phase III trial of HPV type 16 vaccine using virus-like particles of L1 capsid of HPV-16, the incidence was reduced of both HPV-16 infection and HPV-16-related cervical intraepithelial neoplasia. Although preliminary results of immunotherapy clinical trials against CC did not modify the clinical status, they occasionally show improvement of lymphocyte response against HPV. A recent immunotherapy trial using dendritic cells pulsed with HPV-18 E7 oncoprotein as adjuvant resulted in temporal remission and improved performance status in a patient with metastatic CC. New and different vaccine preventive trials against HPV are being put into practice and clinically tested. It is hoped that in the future it may be possible to eradicate cervical cancer. The success of immunotherapy anti-HPV clinical trials in CC patients will be determined at a future time. The scientific basis for the development of papillomavirus prophylactic and therapeutic vaccines against persistent infection and preinvasive-invasive associated cervical lesions along with the present status of immunopreventive and immunotherapy clinical trials against cervical cancer are commented on in this paper. PMID- 15888276 TI - Does a diet rich in dietary fibre really reduce the risk of colon cancer? PMID- 15888277 TI - Present and future of endoscopic ultrasonography. AB - Endoscopic ultrasonography and endoscopic ultrasonography-guided fine needle aspiration are well-established techniques, encompassing a variety of diagnostic and therapeutic applications. Along with traditional indications that constitute everyday clinical practice in all endoscopic ultrasonography centres, new indications are emerging that resemble the continuing research carried on in this field. Some of these are innovative applications, developed by highly experienced endosonographers and with a putative role for clinical practice in the near future. Others are merely experimental applications, carried out on in animal models or in highly selected groups of patients, opening up new fascinating areas of research but not for imminent introduction in clinical practice. The purpose of this review, after summarising the present indications of endoscopic ultrasonography, is to focus on the future applications and try to establish their possible advent, either in the near or in the far future. PMID- 15888278 TI - Co-existence of Helicobacter pylori infection in patients with Familial Mediterranean Fever (FMF) and the effect of Helicobacter pylori on the frequency and severity of FMF attacks. AB - BACKGROUND: The inflammatory reactions both in Familial Mediterranean Fever and in Helicobacter pylori infection have similarities. Whether there is interactions in case of co-existence of both diseases has not been evaluated. AIM.: To evaluate, if there is a significant relation between H. pylori infection and Familial Mediterranean Fever; if H. pylori has an effect on the frequency and severity of Familial Mediterranean Fever attacks; and if eradication treatment has any affects. METHODS: Thirty-two Familial Mediterranean Fever patients were tested for H. pylori infection. Acute phase responses were evaluated and attack frequency and severity were determined in both H. pylori-positive and H. pylori negative groups. Same determinations were done after the eradication treatment in H. pylori-positive patients. Levels of acute phase determinants as well as frequency and severity of attacks were compared in H. pylori-positive and negative groups. RESULTS: C-reactive protein, erythrocyte sedimentation rate, white blood count and fibrinogen levels were significantly (p<0.01) higher during the attacks than before the attacks in all patients. However, there was no difference between the groups. H. pylori-positive patients have a higher frequency and a longer duration of attacks when compared to H. pylori-negative patients before treatment (p<0.05). The frequency was also significantly lower and duration was shorter in patients whose infections were eradicated (p<0.05). CONCLUSION: H. pylori infection was not significantly frequent in our group of Familial Mediterranean Fever patients. H. pylori can decrease both the frequency and the duration of the attacks. Studies that will evaluate the relationship of H. pylori and MEFV gene along with the roles of yet unknown cytokines, which can presumably play a role in the pathogenesis of both diseases, are needed to reach better conclusions. PMID- 15888279 TI - The G2964A polymorphism of the STAT6 gene in inflammatory bowel disease. AB - BACKGROUND AND AIMS: Linkage of inflammatory bowel diseases to chromosome 12p13.2 q24.1 (IBD2) has been confirmed in several genome wide screens. The STAT6 gene is located within this chromosomal region. The transcription factor STAT6 is involved in the regulation of the TH1/TH2 immune response. Increased production of TH1 cytokines is crucial in the pathogenesis of Crohn's disease. PATIENTS AND METHODS: Therefore, we genotyped a single nucleotide polymorphism in the 3' untranslated region of the STAT6 gene (G2964A) in 243 patients with Crohn's disease, 100 patients with ulcerative colitis and 548 healthy controls. RESULTS: In comparison to controls, the G allele and the GG genotype frequencies were significantly increased only in Crohn's disease patients without a variation in the CARD15 gene (p<0.03 and p<0.02, respectively). CONCLUSIONS: Alterations in the STAT6 pathway may play a crucial role in the pathogenesis of distinct subgroups of patients with Crohn's disease. PMID- 15888280 TI - Tumour angiogenesis and mast cell density in the prognostic assessment of colorectal carcinomas. AB - BACKGROUND: Tumour angiogenesis is essential for the growth, invasion and metastasis of solid tumours. There are several lines of evidence that the mast cells play an important role in tumour angiogenesis. AIMS: The study focused to determine the correlation between the microvessel and mast cell densities, and to evaluate whether tumour angiogenesis and mast cell density could predict recurrence following curative surgery in patients with colorectal carcinomas. PATIENTS: Microvessel and mast cell densities were investigated in tumour specimens from 60 patients with colorectal carcinoma. METHODS: Microvessels were stained by immunohistochemical method using a monoclonal antibody anti-CD34. The routine Giemsa blue staining method was used to assess the mast cells. Microvessels and mast cells were counted in a x400 field. RESULTS: The mean microvessel and mast cell counts were higher in patients with recurrence compared with those patients who were disease-free for at least 24 months (p<0.001). The Spearman's correlation coefficient revealed a significant correlation between mast cell and microvessel counts in colorectal carcinomas (r=0.684; p<0.001). Kaplan-Meier plots of survival showed that the high microvessel (>28) and mast cell (>6) counts correlated with a shorter disease-free survival (p=0.0003 and p=0.0013, respectively). Multivariate analysis showed that the depth of penetration (T4 versus T2) (p=0.004), liver metastasis (p=0.04) and microvessel density (p=0.003) were independent predictors of recurrence. In multivariate analysis, mast cell density did not reach significance. CONCLUSIONS: Our results suggest that the microvessel density of the primary tumour may be an important independent predictor of tumour recurrence and time to recurrence in colorectal carcinomas. The significant correlation between mast cell and microvessel counts suggest that the mast cells may have a role in tumour progression via promoting angiogenesis. PMID- 15888281 TI - CTLA4 gene polymorphism in Italian patients with colorectal adenoma and cancer. AB - BACKGROUND AND AIMS: Colorectal cancer is a major health problem. Colonoscopic colorectal cancer screening is cumbersome and expensive. Identification of genetic risk of colorectal cancer may help to select the subjects who could benefit from colonoscopy. The immune system plays a fundamental role in the human environment interaction, and the carcinogenic effects of many environmental factors are mediated by the chronic activation of the immune system in a genetic controlled fashion. Cytotoxic T lymphocyte associated antigen 4 (CTLA4) plays an inhibitory role in regulating lymphocyte functions. The loss of CTLA4 function is responsible for loss of mucosal lymphocyte tolerance. The G allele at position +49 of exon 1 of the CTLA4 gene affects the CTLA4 function. We evaluated in an association study the role of CTLA4 A+49G polymorphism as a risk factor for colorectal neoplasm. PATIENTS AND METHODS: Five hundred and fifty-six patients (male 295; female 261) who underwent colonoscopy at our Centre were enrolled in the study and divided into three groups: Colorectal cancer (132 patients, M/F 68/64, mean age 66+/-11 years); Colorectal adenoma (186 patients, M/F 110/76, mean age 65+/-11 years); Healthy controls (238 patients, M/F 117/121, mean age 63+/-10 years). DNA was extracted from peripheral blood, CTLA4 gene was amplified by using specific primers, and A+49G polymorphism was analysed by restriction enzyme digestion. RESULTS: No statistically significant differences in the genotype distribution among Control and Adenoma groups (p=0.93), Control and Carcinoma groups (p=0.52), and Adenoma and Carcinoma groups (p=0.53) were observed. CONCLUSION: There is no significant correlation between CTLA4 A+49G polymorphism and the risk of colorectal neoplasm among Italian Caucasians. PMID- 15888282 TI - Quantitative treatment of the hyponatremia of cirrhosis. AB - BACKGROUND: Hyponatremia represents a frequent complication of liver cirrhosis, associated with adverse events and death. It is caused either by excessive water retention or solute depletion, or a combination of both. AIMS: To determine the cause of hyponatremia clinically and to examine the usefulness of quantitative calculations of water excess and Na deficit to guide treatment. METHODS: We studied 23 patients with liver cirrhosis and PNa < or =131 meq/L to determine the cause of hyponatremia and results of quantitative treatment. RESULTS: The most frequent cause of hyponatremia was diuretic-induced Na depletion, which occurred in 14 out of 23 instances, while four patients had water excess. Hyponatremia was corrected after a quantitative estimate of the Na deficit or relative water excess by algebraic formulas. The former was quantitatively replenished as 3% NaCl, the latter was excreted with the technique of furosemide-induced diuresis and re-infusion of solute, but not water, losses. After quantitative replacement, there was a significant correlation (R=0.98, P< 0.001) between the Na concentration predicted mathematically and that actually measured. CONCLUSIONS: The hyponatremia of cirrhosis is frequently caused by diuretics. Its treatment can be effectively guided with the aid of quantitative estimates of Na deficit and/or water excess in all instances, although the methods of correction described are indicated in severe clinical conditions. PMID- 15888283 TI - Quality of life in patients with chronic pancreatitis. AB - INTRODUCTION: Health-related quality of life is becoming a major issue in the evaluation of any therapeutic intervention in patients with chronic or hard to cure diseases. AIMS: To assess the quality of life in patients with chronic pancreatitis, the majority of whom have had the disease for a long time, and to evaluate which factors linked to the disease are able to influence the quality of life. SUBJECTS AND METHODS: A total of 190 consecutive patients (157 males, 33 females; mean age 58.6+/-12.7 years, range 18-92 years) with proven chronic pancreatitis were enrolled. The SF-36 questionnaire was used for assessing the health-related quality of life. RESULTS: The z-scores of the eight domains of the patients with chronic pancreatitis were significantly negative indicating an overall impairment of the quality of life when compared to the Italian normative sample. Pancreatic pain was the unique clinical variable able to significantly impair all eight domains of the SF-36, while Wirsung dilation and diabetes were negatively related to some physical and mental domains. The body mass index was the unique variable positively related with some SF-36 domains. CONCLUSIONS: Pain may be considered the most important factor affecting the quality of life of chronic pancreatitis patients; moreover, alimentary and metabolic factors deserve more attention in improving the quality of life of these subjects. PMID- 15888284 TI - Urine diacetylspermine as a novel tumour maker for pancreatobiliary carcinomas. AB - BACKGROUND: Serum carcinoembryonic antigen (highly specific) and carbohydrate antigen 19-9 (highly sensitive) have been used as tumour markers for pancreatobiliary cancers. A novel urine tumour marker, diacetylspermine, was compared with the two conventional serum tumour markers in 125 patients with pancreatobiliary diseases. RESULTS: When the diagnosis of benign or malignant condition was examined, the sensitivity of urine diacetylspermine (75%) was higher than that of serum carcinoembryonic antigen (44%; P=0.048) and the same as that of serum carbohydrate antigen 19-9 (75%). The specificity of urine diacetylspermine (81%) was lower than that of serum CEA (92%) and as high as that of serum carbohydrate antigen 19-9 (80%). The efficiency of urine diacetylspermine (79%) was higher than that of serum carcinoembryonic antigen (74%) and the same as that of serum carbohydrate antigen 19-9 (79%). CONCLUSION: These results suggest that urine diacetylspermine is a marker for pancreatobiliary carcinoma, which is as highly sensitive and specific as serum carbohydrate antigen 19-9. PMID- 15888285 TI - The role of upper endoscopy in identifying oesophageal involvement in patients with oral pemphigus vulgaris. AB - BACKGROUND AND AIMS: The involvement of oesophagus in pemphigus vulgaris is still debated. The aims of this study were to evaluate the prevalence of oesophageal involvement and the gastro-duodenal mucosa appearance before and after high-dose corticosteroid therapy in a group of patients with oral pemphigus vulgaris. METHODS: We prospectively studied 28 consecutive patients with oral pemphigus by oesophageal symptom standardised questionnaire, upper gastro-intestinal endoscopy, exfoliative cytology and histological biopsy. After clinical remission, all patients underwent new endoscopy. RESULTS: The prevalence of oesophageal symptoms was 57.1%. Endoscopic examination revealed oesophageal involvement with different degrees of severity in 67.8% of patients. After corticosteroid therapy, endoscopy showed normal oesophageal-gastro-duodenal mucosa. No examination-related exacerbations of the oesophageal lesions were seen. CONCLUSIONS: The upper gastro-intestinal endoscopic examination, in oral pemphigus vulgaris patients with oesophageal symptoms, is safe in skilled hands technique and a useful diagnostic tool prior to starting therapy. PMID- 15888286 TI - Hepatocellular adenoma: imaging findings and pathological correlation. AB - A 45-year-old man presented with a 1-week history of gastrooesophageal reflux, epigastric discomfort and abdominal pain especially in the right hypochondrium. Abdominal ultrasonography, contrast-enhanced spiral computed tomography and magnetic resonance imaging of the abdomen were performed. Eventually, imaging findings were correlated with histopathological analysis, which confirmed the diagnosis of a 18 cm x 16 cm hepatocellular adenoma. PMID- 15888287 TI - An adult case of acute lymphoblastic leukaemia presenting as hepatic dysfunction. AB - Acute hepatic dysfunction is a rare and often fatal presentation of haematological malignancies. We describe an adult case of acute lymphoblastic leukaemia presenting as an acute hepatitis. Due to the elevation in the patient's transaminases and bilirubin, standard acute lymphoblastic leukaemia induction therapy could not be used. Instead the combination of prednisone and asaparaginase were used to successfully induce remission. PMID- 15888288 TI - Autoimmune pancreatitis. AB - Autoimmune pancreatitis is a type of idiopathic pancreatitis. It is also referred to as sclerosing pancreatitis, lymphoplasmatocytic sclerosing pancreatitis, chronic pancreatitis with irregular stenosis of the main pancreatic duct and as sclerosing pancreatocholangitis. Clinical characteristics of autoimmune pancreatitis are jaundice, abdominal pain, weight loss and diabetes mellitus. Radiologically, there is diffuse enlargement of pancreas with stenosis of pancreatic duct without calcifications in the pancreatic parenchyma. In autoimmune pancreatitis, antibodies against lactoferrin and carbonic anhydrase have been detected, but they are not specific because they are present in some other autoimmune diseases too. Also in autoimmune pancreatitis, there are increased levels of gammaglobulins and characteristically high titres of IgG4, which are a subtype of IgG. Autoimmune pancreatitis is usually treated successfully by prednisolone. PMID- 15888289 TI - Giardia lamblia: expression of alkaline phosphatase activity in infected rat intestine. AB - Alkaline phosphatase (IAP) is a marker of intestinal microvillus membrane. Changes in IAP activity have been studied as a function of Giardia lamblia (G. lamblia) infection using rat as the experimental model. At day 11 and 15 post infection, enzyme activity was reduced (p<0.01) compared to controls. The enzyme levels were essentially similar to control values by day 30 post-infection. Analysis of the enzyme activity in cell fractions across crypt-villus axis revealed a marked decrease in enzyme activity in the villus tip and mid villus regions but a considerable increase (p<0.01) in enzyme activity in the crypt base of 11 day post-infected animals compared to that in controls. The observed changes in IAP activity were confirmed by assaying the enzyme activity in acrylamide gels using bromo-chloro-indolyl phosphate staining under non denaturing conditions. These findings indicate differential changes across the crypt-villus axis, but impaired alkaline phosphatase levels in G. lamblia infected rat intestine. PMID- 15888290 TI - Trypanosoma cruzi: effects of repetitive stress during the development of experimental infection. AB - Activation of the hypothalamus-pituitary-adrenal axis plays a major role in the suppression of the immune system. We have investigated the effects of repetitive stress on Wistar rats infected with the Y strain of Trypanosoma cruzi and a control group that underwent stressor stimuli by exposure to ether vapor for one minute twice a day. Repetitive stress resulted in an elevated number of circulating parasites accompanies by deep tissue disorganization, and cardiac histopathological alterations. The infected and stressed group displayed a decrease in body weight, and an increased parasite burden in heart tissue, and adrenal glands. Histological analysis of the heart also showed a moderate to severe diffused mononuclear inflammatory process. These results suggest that repetitive stress could be considered an important factor during development of experimental Chagas' disease, enhancing pathogenesis through disturbance of the host's immune system. PMID- 15888291 TI - Leishmania chagasi: the alpha-tubulin intercoding region results in constant levels of mRNA abundance despite protein synthesis inhibition and growth state. AB - The intercoding regions of many Leishmania sp. genes have been implicated in the regulation of mRNA processing, stability, and translation. Herein we show that the intercoding region of the Leishmania chagasi alpha-tubulin gene (alpha-TUB) confers stable beta-galactosidase (beta-GAL) reporter mRNA levels during promastigote growth and development in vitro and during protein synthesis inhibition. The abundance of both endogenous alpha-TUB mRNA and beta-GAL mRNA from a beta-GAL coding region situated upstream of the alpha-TUB intercoding region did not change significantly as promastigotes grew from logarithmic to stationary phase in vitro and the half-life of the beta-GAL mRNA remained constant. The abundance of both the endogenous alpha-TUB and the beta-GAL mRNA increased by less than 2-fold after protein synthesis inhibition corresponding to a moderate increase in mRNA half-life. These data suggest that the alpha-TUB intercoding region is an excellent control for the study of the regulation of other differentially expressed genes. PMID- 15888292 TI - Trichomonas vaginalis: intrastrain polymorphisms within the ribosomal intergenic spacer do not correlate with clinical presentation. AB - Trichomoniasis presents a broad spectrum of clinical patterns ranging from asymptomatic to severe vaginitis and cervicitis. Despite its importance, very little is known about the genetic relatedness of its causative agent, Trichomonas vaginalis, and the clinical phenotypes. To address this question, analysis of restriction length polymorphism (RFLP) within the intergenic spacer of the ribosomal DNA (IGS) from 60 clinically defined isolates of T. vaginalis was performed. This is the first description of the IGS polymorphism of T. vaginalis. As expected, a considerable number of patients were asymptomatic (28%) while only 12% presented both leukorrhea and macular colpitis, the most evident symptoms of trichomoniasis. The IGS-RFLP with the use of eight restriction enzymes showed absence of correlation between the genetic relatedness of the isolates and symptomatology. Further studies are necessary to evaluate the importance of the IGS polymorphism to the parasite virulence and clinical phenotype. PMID- 15888293 TI - Acanthamoeba healyi: molecular cloning and characterization of a coronin homologue, an actin-related protein. AB - Coronin, described in organisms from yeasts to humans, has been found to be involved in various actin-associated activities. It has yet to be described in Acanthamoeba, medically significant as the causative agent of granulomatous amebic encephalitis and amoebic keratitis and used extensively in actin-related studies. We isolated and characterized a cDNA encoding a coronin-like protein in A. healyi by sequence analysis and demonstrated intracellular localization of the gene product by transient transfection. Named Ahcoronin, the gene is composed of 454 amino acids which contain the characteristic WD repeats of coronin and coronin-like proteins. The C-terminal region of the gene was also predicted to have a high tendency of forming a coiled-coil, another structural characteristic of coronin. The gene showed a 50% homology to coronins. Ahcoronin occurs as a single copy and expressed as a transcript of approximately 1.4kb in A. healyi. Results of transfection showed that Ahcoronin was localized in the cell's periphery and in the leading edge consistent to that of actin. The fusion protein has also been observed to localize around phagocytic cups but was disassembled later during phagocytosis. Sequence analysis of Ahcoronin homologue of A. healyi showed numerous potential for further studies and is sure to contribute in the growing interest toward the properties and functions of coronin and coronin-like proteins. PMID- 15888294 TI - Isolation and characterization of a cDNA encoding a serine proteinase from the root-knot nematode Meloidogyne incognita. AB - This report describes the first serine proteinase gene isolated from the sedentary nematode Meloidogyne incognita. Using degenerate primers, a 1372bp cDNA encoding a chymotrypsin-like serine proteinase (Mi-ser1) was amplified from total RNA of adult females by RT-PCR and 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of Mi-ser1 encoded a putative signal peptide and a prodomain of 22 and 33 amino acids, respectively, and a mature proteinase of 341 amino acids with a predicted molecular mass of 37,680Da. Sequence identity with the top serine proteinases matches from the databases ranged from 23 to 27%, including sequences from insects, mammals, and other nematodes. Southern blot analysis suggested that Mi-ser1 is encoded by a single or few gene copies. The pattern of developmental expression analyzed by Northern blot and RT-PCR indicated that Mi-ser1 was transcribed mainly in females. The domain architecture composed of a single chymotrypsin-like catalytic domain and the detection of a putative signal peptide suggested a digestive role for Mi-ser1. PMID- 15888295 TI - Toxocara canis: search for a potential drug amongst beta-carboline alkaloids--in vitro and mouse studies. AB - The goal of this study was to search for new treatments for Toxocara canis using both in vitro and in vivo experiments. We specifically looked for a treatment for T. canis larva migrans, and examined beta-carboline alkaloids (17 compounds) with various structural modifications, both in in vitro and in vivo experiments. In the in vitro experiments, screening for nematocidal activity on the T. canis second stage larvae, cytotoxic activity, and immune activity in the host were undertaken. Compound 17 was selected, as it exhibited nematocidal activity for T. canis larvae and did not have any cytotoxic or immunosuppressive activity in the host. The effectiveness of compound 17 was then examined using T. canis larvae infected mice in in vivo experiments. To evaluate the anthelmintic effect, the relative mobility value for the larvae was examined in addition to the number of larvae in the brain, skeletal muscle, and liver. Compound 17 was also examined in both free and liposome-entrapped (LE) forms. Polyethylene glycol (PEG)-LE compound 17 showed an anthelmintic effect in which the number of larvae in the brain was decreased compared free albendazole. PEG-LE compound 17 also effectively suppressed the mobility of the larva in brain and skeletal muscle. The experimental procedure employed assisted in the discovery of this potential candidate and is a promising approach for finding alternative therapeutic regimens for T. canis larva migrans. PMID- 15888296 TI - Gnathostoma binucleatum: excretion-secretion antigen analysis obtained from advanced third-stage larvae in in vitro culture. AB - The paper describes an introductory characterization of antigenic stimulation of excretion-secretion products (ESP) of Gnathostoma binucleatum advanced third stage larvae cultured in vitro and proteinases present in this products. Excretory and secretory proteins were obtained after 10 larvae were maintained in 5% CO(2) RPMI medium. The supernatant was collected each week for two months. The proteins were dialyzed, concentrated, and separated in 10% SDS-PAGE gels under reducing conditions and transferred to nitrocellulose paper for immunoblot analyses. G. binucleatum immunized mice serum was used to determine protein antigenicity. Four proteins of 40, 80, 120, and 208 kDa persisted for two months and three proteins, 80, 120, and 208 kDa were recognized for antibodies of mice. In SDS-PAGE gelatin substrate gels ESP resolved as two proteins with molecular weight of 80 and 208 kDa that were sensitive to a metalloproteinase inhibitor, and thus it may be inferred that they might be used for diagnosis of gnathostomiasis. PMID- 15888297 TI - Morphological characterization of the ovary and oocytes vitellogenesis of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae). AB - This study presents the morphology of the ovary, as well as the process of the vitellogenesis in oocytes of the tick Rhipicephalus sanguineus. The ovary of these individuals is of the panoistic type; therefore, it lacks nurse cells. This organ consists of a single tubular structure, continuous, and composed of a wall formed by small epithelial cells with rounded nuclei which delimit the lumen. The oocytes in the different developmental stages in this tick species were classified into five stages (I-V). They remain attached to the ovary during vitellogenesis by a cellular pedicel and afterwards the mature oocytes (stage V) are released into the ovary lumen. PMID- 15888298 TI - Schistosoma mansoni: ferredoxin-NADP(H) oxidoreductase and the metabolism of reactive oxygen species. AB - Mitochondrial-type ferredoxin-NADP(H) oxidoreductases (FNR) catalyze the electron transport between NADPH and substrates such as ferredoxins. Even though enzymes belonging to this family are present in several organisms, including prokaryotes, their biological function is not clearly understood. In a previous work, we reported the existence of a mitochondrial-type FNR in the trematode Schistosoma mansoni (SmFNR). This enzyme conferred tolerance to oxidative stress conditions when tested in an heterologous system. In this work, we demonstrate that the SmFNR can be imported to mitochondria in mammal cells and show that its expression is induced in parasite cultures by reactive oxygen species (ROS). The results reported herein give further support to the involvement of SmFNR in ROS metabolism. PMID- 15888299 TI - Effect of a juvenile hormone analogue on Leishmania amazonensis and Leishmania braziliensis. PMID- 15888300 TI - Toxoplasma gondii binds human lactoferrin but not transferrin. PMID- 15888301 TI - Persistence and coexistence of engineered baculoviruses. AB - Baculoviruses, and in particular, the nucleopolyhedroviruses infect a wide range of arthropod hosts and have the potential to be used as biopesticides. However, one of the major drawbacks with these pathogens as biocontrol agents is that they have a slow response time. Alterations to the speed of kill and pathogen life history characteristics can influence the competitive outcome and persistence between wildtype and modified strains. Here, we explore, theoretically, how life history modifications of pathogens can affect the epidemiology and ecology of strain coexistence. In particular, we show how under simple mass action disease transmission, life-history difference between strains are insufficient to allow coexistence. Additional heterogeneities in transmission are shown to be necessary to facilitate coexistence of wildtype and modified pathogen strains. We also illustrate how the patterns of infectivity of wildtype and modified strains can also affect long-term coexistence, and argue that appropriate assessment of genetic modifications must be presented in terms of relevant ecological theory. PMID- 15888302 TI - Measuring diversity from dissimilarities with Rao's quadratic entropy: are any dissimilarities suitable? AB - Rao has developed quadratic entropy to measure diversity in a set of entities divided up among a fixed set of categories. This index depends on a chosen matrix of dissimilarities among categories and a frequency distribution of these categories. With certain choices of dissimilarities, this index could be maximized over all frequency distributions by eliminating several categories. This unexpected result is radically opposite to those obtained with usual diversity indices. We demonstrate that the elimination of categories to maximize the quadratic entropy depends on mathematical properties of the chosen dissimilarities. In particular, when quadratic entropy is applied to ultrametric dissimilarities, all categories are retained in order to reach its maximal value. Three examples, varying from simple one-dimensional to ultrametric dissimilarity matrices, are provided. We conclude that, as far as diversity measurement is concerned, quadratic entropy is most relevant when applied to ultrametric dissimilarities. PMID- 15888303 TI - Families of discrete kernels for modeling dispersal. AB - Integer lattices are important theoretical landscapes for studying the consequences of dispersal and spatial population structure, but convenient dispersal kernels able to represent important features of dispersal in nature have been lacking for lattices. Because leptokurtic (centrally peaked and long tailed) kernels are common in nature and have important effects in models, of particular interest are families of dispersal kernels in which the degree of leptokurtosis can be varied parametrically. Here we develop families of kernels on integer lattices with several important properties. The degree of leptokurtosis can be varied parametrically from near 0 (the Gaussian value) to infinity. These kernels are all asymptotically radially symmetric. (Exact radial symmetry is impossible on lattices except in one dimension.) They have separate parameters for shape and scale, and their lower order moments and Fourier transforms are given by simple formulae. In most cases, the kernel families that we develop are closed under convolution so that multiple steps of a kernel remain within the same family. Included in these families are kernels with asymptotic power function tails, which have provided good fits to some observations from nature. These kernel families are constructed by randomizing convolutions of stepping-stone kernels and have interpretations in terms of population heterogeneity and heterogeneous physical processes. PMID- 15888304 TI - The dynamics of adaptation: an illuminating example and a Hamilton-Jacobi approach. AB - Our starting point is a selection-mutation equation describing the adaptive dynamics of a quantitative trait under the influence of an ecological feedback loop. Based on the assumption of small (but frequent) mutations we employ asymptotic analysis to derive a Hamilton-Jacobi equation. Well-established and powerful numerical tools for solving the Hamilton-Jacobi equations then allow us to easily compute the evolution of the trait in a monomorphic population when this evolution is continuous but also when the trait exhibits a jump. By adapting the numerical method we can, at the expense of a significantly increased computing time, also capture the branching event in which a monomorphic population turns dimorphic and subsequently follow the evolution of the two traits in the dimorphic population. From the beginning we concentrate on a caricatural yet interesting model for competition for two resources. This provides the perhaps simplest example of branching and has the great advantage that it can be analyzed and understood in detail. PMID- 15888305 TI - Threshold policies control for predator-prey systems using a control Liapunov function approach. AB - The stability of predator-prey models, in the context of exploitation of renewable resources, subject to threshold policies (TP) is studied in this paper using the idea of backstepping and control Liapunov functions (CLF) well known in control theory, as well as the concept of virtual equilibria. TPs are defined and analysed for different types of one and two species predator-prey models. The models studied are the single species Noy-Meir herbivore-vegetation model, in a grazing management context, as well as the Rosenzweig-MacArthur two species predator-prey model, in a fishery management context. TPs are shown to be versatile and useful in managing renewable resources, being simple to design and implement, and also yielding advantages in situations of overexploitation. PMID- 15888307 TI - Oxidative cascades: a facile biosynthetic strategy for the assembly of complex molecules. AB - Electron-rich aromatic compounds undergo a facile tandem reaction sequence involving an iterative two-electron oxidation/aromatization. This review will describe the application of this motif to the synthesis of dimethylbenzimidazole, pyoverdine, actinomycin, cystodytin, pyrroloquinoline quinone, and the cataract pigment. PMID- 15888308 TI - Phosphoryl transfer by aminoglycoside 3'-phosphotransferases and manifestation of antibiotic resistance. AB - Transfer of the gamma-phosphoryl group from ATP to aminoglycoside antibiotics by aminoglycoside 3'-phosphotransferases is one of the most important reactions for manifestation of bacterial resistance to this class of antibiotics. This review article surveys the latest structural and mechanistic findings with these enzymes. PMID- 15888309 TI - Structure and mechanism of tryptophylquinone enzymes. AB - Tryptophylquinone cofactors are formed by posttranslational modifications that result in the incorporation of two oxygens into a tryptophan side chain, and the covalent cross-linking of that side chain to another amino acid residue. Tryptophylquinone enzymes catalyze the oxidative deamination of primary amines, and utilize other redox proteins as electron acceptors. Mechanistic and structural studies of these enzymes are providing insight into how these enzymes utilize these highly reactive protein-derived quinones in a controlled manner to facilitate biologically important catalytic and electron transfer reactions. PMID- 15888310 TI - Mechanism and applications of phosphite dehydrogenase. AB - Phosphite dehydrogenase catalyzes the NAD+-dependent oxidation of hydrogen phosphonate (common name phosphite) to phosphate in what amounts to a formal phosphoryl transfer reaction from hydride to hydroxide. This review places the enzyme in the context of phosphorus redox metabolism in nature and discusses the results of mechanistic investigations into its reaction mechanism. The potential of the enzyme as a NAD(P)H cofactor regeneration system is discussed as well as efforts to engineer the cofactor specificity of the protein. PMID- 15888311 TI - Experimental observation of thiamin diphosphate-bound intermediates on enzymes and mechanistic information derived from these observations. AB - Thiamin diphosphate (ThDP), the vitamin B1 coenzyme, is an excellent representative of coenzymes, which carry out electrophilic catalysis by forming a covalent complex with their substrates. The function of ThDP is to greatly increase the acidity of two carbon acids by stabilizing their conjugate bases, the ylide/C2-carbanion of the thiazolium ring and the C2alpha-carbanion (or enamine) once the substrate binds to ThDP. In recent years, several ThDP-bound intermediates on such pathways have been characterized by both solution and solid state (X-ray) methods. Prominent among these advances are X-ray crystallographic results identifying both oxidative and non-oxidative intermediates, rapid chemical quench followed by NMR detection of a several intermediates which are stable under acidic conditions, and circular dichroism detection of the 1',4' imino tautomer of ThDP in some of the intermediates. Some of these methods also enable the investigator to determine the rate-limiting step in the complex series of steps. PMID- 15888312 TI - The enzymes of sialic acid biosynthesis. AB - The sialic acids are a family of nine carbon alpha-keto acids that play a wide variety of biological roles in nature. In mammals, they are found at the distal ends of cell surface glycoconjugates, and thus are major determinants of cellular recognition and adhesion events. In certain strains of pathogenic bacteria, they are found in capsular polysaccharides that mask the organism from the immune system by mimicking the exterior of a mammalian cell. This review outlines recent developments in the understanding of the two main enzymes responsible for the biosynthesis of the sialic acid, N-acetylneuraminic acid. The first, a hydrolyzing UDP-N-acetylglucosamine 2-epimerase, generates N-acetylmannosamine and UDP from UDP-N-acetylglucosamine. The second, sialic acid synthase, generates either N-acetylneuraminic acid (bacteria) or N-acetylneuraminic acid 9-phosphate (mammals) in a condensation reaction with phosphoenolpyruvate. An emphasis is placed on an understanding of the mechanistic and structural features of these enzymes. PMID- 15888313 TI - Transglycosylation: a mechanism for RNA modification (and editing?). AB - The vast majority of the ca. 100 chemically distinct modified nucleosides in RNA appear to arise via the chemical transformation of a genetically encoded nucleoside. Two notable exceptions are queuosine and pseudouridine, which are incorporated into tRNA via transglycosylation. Transglycosylation is an extremely efficient process for incorporating highly modified bases such as queuine into RNA. Transglycosylation is also a requisite process for "isomerizing" an N nucleoside into a C-nucleoside as is the case for pseudouridine formation. Finally, transglycosylation is an attractive possibility for certain RNA editing events (e.g., pyrimidine to purine conversions) that cannot occur via the known, more straightforward enzymatic reactions (e.g., deaminations). This review discusses what is known about the mechanisms of transglycosylation for the queuine and pseudouridine RNA modifications and will speculate about a potential role for transglycosylation in certain RNA editing events. PMID- 15888314 TI - Making ends meet in old age: DSB repair and aging. AB - Accumulation of somatic mutations has long been considered as a major cause of aging and age-related diseases such as cancer. Genomic rearrangements, which arise from aberrant repair of DNA breaks, are the most characteristic component of the mutation spectra in aging cells and tissues. The studies conducted in the past few years provide further support for the role of DNA double-strand break (DSB) repair in aging and cellular senescence. Evidence was obtained that in addition to accumulation of mutations the efficiency and fidelity of repair declines with age. We propose that DNA damage and age-related decline of DNA repair form a vicious cycle leading to amplification of damage and progression of aging, and discuss a hypothesis on how the interplay between the two pathways of DSB repair, homologous recombination and nonhomologous end joining, may contribute to the aging process. PMID- 15888315 TI - Increased expression of 14-3-3varepsilon protein in intrinsically aged and photoaged human skin in vivo. AB - Skin aging is a complicated process associated with the passage of time and environmental exposure, especially to UV light. This aging phenomenon is related to alterations in various cellular mechanisms, such as changes in apoptosis, perturbations to cellular signaling, and an increased genetic instability. In this study, we investigated changes of proteins involved in intrinsic aging by the proteomic analysis of human sun-protected (upper inner arm) young and aged dermis. One of the proteins upregulated in aged dermis was identified as 14-3 3epsilon. This protein is an isoform of 14-3-3 protein, which is involved in cellular processes like signal transduction, cell cycle arrest, and apoptosis. 14 3-3epsilon is consistently found to be upregulated in the sun-protected dermis of aged skin, by Western blotting and immunohistochemical staining. In addition, we demonstrate that the expression of 14-3-3epsilon is further upregulated in the sun-exposed (photodamaged) dermis, and that the UV irradiation of young skin significantly upregulates 14-3-3epsilon in vivo. Our results suggest the possibility that the cellular processes related to 14-3-3epsilon protein play an important role in the photoaging and intrinsic aging of human skin. PMID- 15888316 TI - Effect of oxidative stress on translocation of DAF-16 in oxygen-sensitive mutants, mev-1 and gas-1 of Caenorhabditis elegans. AB - Mutations in the mev-1 and gas-1 genes of the nematode Caenorhabditis elegans render animals hypersensitive to oxygen and paraquat, and lead to premature aging. We show that both mutants overproduce superoxide anion in isolated sub mitochondrial particles, which probably explains their hypersensitivity to oxidative stress. The daf-16 gene encodes a fork-head transcription factor that is negatively regulated by an insulin-signaling pathway. In wild-type animals, the DAF-16 protein normally resides in the cytoplasm and only becomes translocated to nuclei upon activating stimuli such as oxidative stress. Conversely, DAF-16 resides constitutively in the nuclei of mev-1 and gas-1 mutants even under normal growth conditions. Supplementation of the antioxidant coenzyme Q(10) reversed this nuclear translocation of DAF-16. Since both gas-1 and mev-1 encode subunits of electron transport chain complexes, these data illustrate how mitochondrial perturbations can impact signal transduction pathways. PMID- 15888317 TI - The p38 signal transduction pathway participates in the oxidative stress-mediated translocation of DAF-16 to Caenorhabditis elegans nuclei. AB - Much attention has focused on the insulin-like signaling pathway in Caenorhabditis elegans because of its pivotal role in life-span determination and oxidative stress resistance. The daf-16 gene encodes a fork-head transcription factor that is negatively regulated by this insulin-signaling pathway. The DAF-16 protein is translocated to the nucleus when animals were subjected to oxidative stress in the form of paraquat. This oxidative stress-mediated translocation was blocked by mutation of the p38-related sek-1 (MAPKK) mutant and DAF-16 instead remained cytoplasmic. The fact that DAF-16 translocation by oxidative stress is epistatic to sek-1 suggests that oxidative stress mediates regulation of DAF-16 through activating the p38 signal transduction pathway upstream of daf-16 so as to mobilize DAF-16 to the nucleus and activate transcription. PMID- 15888318 TI - Dietary restriction counteracts age-related changes in cholesterol metabolism in the rat. AB - The effects of ageing on the metabolism of cholesterol were examined in three different organs (liver, aorta and brain) of 6-, 12- and 24-month-old male Sprague-Dawley rats. Ageing was associated with a significant increase in intracellular cholesterol esters in all three organs. Steady state mRNA levels of multidrug resistance protein (MDR) and acylCoA:cholesterol acyl transferase (ACAT), enzymes involved in cholesterol import and esterification, were also increased. By contrast, expression of mRNA for neutral cholesterol ester hydrolase (nCEH) and caveolin-1, proteins involved in cholesterol ester hydrolysis and export, were significantly reduced. Dietary restriction is the only intervention shown to extend lifespan and retard age-related declines in function in mammals. To further explore the possible correlation between changes in cholesterol esterification and ageing, we analysed cholesterol metabolism in liver, aorta, and brain of aged rats exposed to two dietary restriction regimens: intermittent (alternate-day) fasting (IF) and food intake restriction (60% of ad libitum feeding). Both dietary regimens attenuated the age-related changes in cholesterol esters and in the expression of genes involved in cholesterol metabolism. These results provide evidence that distinctive age-associated changes in intracellular cholesterol metabolism occur in rats. Furthermore, these modifications can be partially reversed by dietary restriction, a condition known to affect the ageing process. Age-related changes in cholesterol metabolism may play a role in triggering and/or aggravating senescence-related disorders characterized by altered cholesterol homeostasis. PMID- 15888319 TI - Effect of treatment with dexamethasone on protein intake in adult and old Lou/c/jall rats. AB - A deleterious decrease of protein intake had been evidenced in Lou/c/jall rats during ageing. This result could be induced by an impaired regulation of feeding behaviour. Glucocorticoids inducing specific amino-acid needs for gluconeogenesis and for the synthesis of inflammatory proteins by the liver, we investigated the age-related effect of a 4-days treatment with dexamethasone (DEX) on caloric and protein intake. Males and females aged 7, 19, 25 and 31 months received 573.6 +/- 65.6 microg/(kg day) of dexamethasone via the drinking water. Body weight (BW), caloric and macronutrients intakes were monitored during treatment and during 10 days after the treatment. A strong hypophagia was seen during treatment in all groups, which was mainly due to a decrease in fat intake. In the same time, rats maintained their protein intake so that protein became the main macronutrient of the diet in most of the groups. However, older males showed a lesser efficiency in adjusting their diet. These results are in agreement with previous data obtained in a protein deprivation study. They lead to the conclusion that the loss of appetite for protein in old age probably does not reflect a loss of ability to choose the needed amount of protein. We can hypothesise that the decrease of protein intake in old rats could be due to some inadequacy of casein to the metabolic requirement of aged animals. PMID- 15888320 TI - Analysis of UV-induced damage and repair in young and senescent human dermal fibroblasts using the comet assay. AB - A major cause of ageing is thought to be the accumulation of damage to macromolecules. Accumulation to DNA damage in cells therefore presupposes that aged cells are unable to repair this damage. We have used the in vitro model of cellular ageing to test the idea that senescent cells are deficient in some aspect of DNA repair. Using the alkaline single cell gel electrophoresis assay (comet assay), we have determined the responses of young and senescent human dermal fibroblasts to DNA damage caused by exposure to UVC light. At low doses of UVC, senescent cells generate smaller comets than young cells whilst at medium doses the situation is reversed. At high doses, young and senescent cells respond similarly to one another. Time course experiments revealing repair of DNA damage show that senescent cells generate larger comets than young cells at early stages of repair suggesting that either senescent cells bear more damage per genome than do young cells or that senescent cells are more efficient at excising bulky adducts from DNA. Cells maintained in low levels of serum irrespective of age are less able to repair DNA damage compared with cells maintained in high levels of serum, and furthermore young and senescent cells maintained in high levels of serum are equally able to repair DNA damage. Our data, therefore, reveal both age dependent and age-independent responses to UV-induced DNA damage. Use of the comet assay highlights the heterogeneity of cellular responses to genotoxic stress. PMID- 15888321 TI - Effects of age on immunohistochemical changes in the mouse hippocampus. AB - We investigated the age-related changes in neuronal cell death and synaptophysin of the hippocampal CA1 sector in mice using immunohistochemistry. Microtubule associated protein 2a, b (MAP2) and synaptophysin immunoreactivity was measured in 2-, 8-, 18-, 42- and 59-week-old mice. MAP2 immunoreactivity was unchanged in the hippocampal CA1 sector up to 42 weeks after birth. In 59-week-old mice, however, a significant decrease in MAP2 immunoreactivity was observed in the hippocampal CA1 sector. Total number of synaptophysin-positive boutons was also unchanged in the hippocampal CA1 sector up to 42 weeks of birth. In 59-week-old mice, however, a significant increase in synaptophysin-positive boutons was observed in the hippocampal CA1 sector. These results demonstrate that dendrites and axons in the hippocampal CA1 neurons are particularly susceptible to ageing processes. In contrast, a marked increase in synaptophysin-positive boutons was found in the hippocampal CA1 sector of aged mice. These findings suggest that increase in synaptophysin-positive boutons may play a role in the maintenance of the structural components in the hippocampal CA1 sector of aged mice although most postsynaptic CA1 pyramidal neurons are generated. Thus, our findings provide further valuable information on age-related neurodegeneration and deficits in hippocampus-dependent memory and synaptic plasticity. PMID- 15888322 TI - Age and gender effects on apoptosis in the human coronary arterial wall. AB - The relationship between aging and apoptosis remains unclear. We wondered whether apoptosis could be enhanced in arterial aging in the absence of overt or advanced arterial disease. Apoptosis-related proteins were investigated using three methods: TdT-mediated dUTP digoxigenin nick end labeling (TUNEL) technique, active cysteine-dependant aspartate specific proteases (caspase)-3 and poly ADP ribose polymerase (PARP) in coronary arteries of human subjects ranging from 25 to 92 years. We found no significant correlation between age and the apoptotic index using the three methods. The percentage of active caspase-3 positive cells was found to be significantly higher in men than in women (9.11 +/- 12.3 cells/mm(2) versus 2.01 +/- 4.55 cells/mm(2), respectively, p = 0.017). These sex related differences did not reach statistical significance using TUNEL (9.93 +/- 17 and 2.61 +/- 4.58 cells/mm(2), p = 0.32) and PARP methods (3.42 +/- 7.74 and 0.86 +/- 0.95 cells/mm(2), p < 0.49). This is the first report of detection of apoptotic cells in the human arterial wall in adult subjects free from arterial diseases. Apoptosis is an attractive hypothesis to account for organ aging, but our study suggests that apoptosis is not a key factor in aging of the arterial wall. PMID- 15888323 TI - Lagomorphs (rabbits, pikas and hares) do not use telomere-directed replicative aging in vitro. AB - Telomere shortening is used for replicative aging in primates and ungulates but not rodents. We examined telomere biology in rabbits to expand the comparative biology of telomere-directed replicative senescence within mammals. The order Lagomorpha consists of two families; Leporidae and Ochotonidae. We examined telomere biology in species representing three leporid genera (European White Rabbit, Black-tailed Jack Rabbit, and Swamp Rabbit) and the monotypic ochotonid genus (North American Pika). Of the leporids one species was a laboratory strain and the others were wild caught. The leporids neither exhibited cellular senescence after sustained periods in culture nor displayed detectable telomerase activity. Continued culture was possible because of their extremely long telomeric arrays. Immunofluorescence showed robust telomere signals at chromosome ends and significant internal chromosomal staining in some instances. Pika was unique in displaying endogenous telomerase activity throughout time in culture. These results show that it is unlikely that lagomorphs use telomere shortening and replicative senescence as a tumor protective mechanism. PMID- 15888324 TI - Hepatic gene and protein expression of primary components of the IGF-I axis in long lived Snell dwarf mice. AB - Recent evidence indicates that the GH/IGF-I axis plays a key role in the control of aging and longevity. To better understand this biological relationship we examined the mRNA and corresponding protein levels of primary IGF-I axis genes in the livers of young and aged long-lived Snell dwarf mice relative to their age matched controls. We demonstrated that the level of IGF-I and ALS mRNAs is dramatically decreased in both young and aged dwarf livers, transcripts encoding IGF-IR and IGFBP-I are elevated in young dwarfs, but normalize to control levels in aged dwarf livers while transcripts encoding IGFBP-3 are elevated only in aged controls. Interestingly, regulation at the protein level of several IGF-I axis components in the Snell dwarf appears to involve both altered gene expression and post-translational regulation. In this study, we reveal both concordant and discordant relationships between mRNA and protein levels for particular components of the IGF-I axis, illustrating that some of these gene products are not solely regulated by transcriptional mechanisms. These results are consistent with a delay in the molecular maturation of the IGF-I axis in dwarf livers, suggesting the preservation of some neonatal characteristics in young adult and aged dwarf livers. Our studies provide gene expression and protein abundance profiles for components of IGF-I axis that are distinguishing characteristics of both young and aged dwarf mice, and suggest that delayed development of the IGF-I axis in the young adult Pit1(dw/dwJ) dwarf liver may play an important role in the endocrine regulation of mammalian longevity. PMID- 15888325 TI - Longevity-associated NADH dehydrogenase subunit-2 polymorphism and serum electrolyte levels in middle-aged obese Japanese men. AB - Mitochondrial DNA 5178 cytosine/adenine polymorphism, which is also called NADH dehydrogenase subunit-2 237 leucine/methionine (ND2-237 Leu/Met) polymorphism is associated with Japanese longevity. This polymorphism is widely associated with blood pressure, serum lipid levels, hematological parameters, intraocular pressure, and serum protein fraction levels. However, there have been no reports on the association between ND2-237 Leu/Met polymorphism and serum electrolyte levels. To investigate this relationship, we performed an association study in 321 healthy middle-aged Japanese men. Crude data showed that serum sodium levels and serum chloride levels were significantly lower in men with ND2-237 Met than in those with ND2-237 Leu (P = 0.021 and 0.003, respectively). Cigarette consumption and body mass index were significantly and positively associated with serum chloride levels (P = 0.002 and 0.008, respectively) and hemoglobin levels were significantly and negatively associated with them (P = 0.007) in ND2-237 Leu genotypic men. In men with ND2-237 Met, only hemoglobin levels were significantly and negatively associated with serum chloride levels (P = 0.025). After adjusting for covariates, only in male obese (body mass index> or =25) subjects, serum sodium and chloride levels remained significantly lower, and serum calcium levels appeared to be significantly higher in ND2-237 Met than in ND2-237 Leu (P = 0.013, <0.001, and 0.046, respectively). Longevity-associated NADH dehydrogenase subunit-2 polymorphism may influence serum electrolyte levels in middle-aged obese Japanese men. PMID- 15888326 TI - Ageing alters the production of nitric oxide and prostanoids after IL-1beta exposure in mesenteric resistance arteries. AB - We aimed to analyse age influence on the production of inflammatory mediators from inducible isoforms of nitric oxide synthase (iNOS) and cyclooxygenase (COX 2) in rat mesenteric resistance arteries (MRA). The second and/or third branches of MRA from young (3-month-old) and old (22-month-old) male Sprague-Dawley rats were incubated in culture medium with or without interleukin-1ss (IL-1ss; 10 ng/ml, 14 h). IL-1ss did not modify endothelial NOS (eNOS) expression or endothelial cell distribution. However, IL-1ss increased nitrite production and iNOS expression in endothelial and smooth muscle cells more in arteries from young than from old rats. IL-1ss also increased PGI(2) levels and COX-2 expression in the three layers of the vascular wall. Ageing did not affect COX-2 expression but did increase TXA(2) and PGF(2alpha) levels. The maximum contraction to phenylephrine was increased in arteries from old rats after IL-1ss treatment. Inhibition of iNOS and COX-2 with 1400 W and NS398, respectively, abolished the differences in phenylephrine contraction. In conclusion, IL-1ss induced an inflammatory response in MRA with associated increases in iNOS and COX 2 expression. The lower increase in nitrite production from iNOS together with a greater contractile prostanoid production in the old rats would be responsible for the increase observed in their contraction to phenylephrine after IL-1 ss treatment. PMID- 15888327 TI - Reciprocal age related change in natural killer cell receptors for MHC class I. AB - Natural killer (NK) cells are essential for healthy aging. NK cell activation is controlled by MHC class I-specific CD94/NKG2 receptors and killer immunoglobulin like receptors (KIR). To assess NK cytotoxic function in isolation from MHC receptor engagement, we measured the ability of purified NK cells to kill mouse P815 target cells in the presence of anti-CD16 mAb. CD16-mediated cytotoxicity did not change with age, indicating that NK activation and cytotoxic granule release remained functional. We then investigated MHC class I receptor expression on NK cells. There was an age related decrease in CD94 and NKG2A expression and a reciprocal age related increase in KIR expression. NKG2A expression also declined with age on CD56(+) T cells. CD94/NKG2A receptor function was proportional to expression, indicating that NK cell inhibitory signaling pathways were intact. NKG2A and KIR expression were complementary, suggesting that CD94/NKG2A function could substitute for inhibitory KIR function during polyclonal NK cell development in both young and elderly adults. The distinct roles of CD94/NKG2A and KIR receptors suggest that shifting MHC class I receptor expression patterns reflect age related changes in NK cell and CD56(+) T cell turnover and function in vivo. PMID- 15888328 TI - Hematopoietic malignancies associated with increased Stat5 and Bcl-x(L) expressions in Ink4a/Arf-deficient mice. AB - The INK4a/ARF locus, which encodes the two distinct proteins p16(INK4a) and p14(ARF), is frequently altered in various hematological malignancies as well as in other types of cancers in humans. In this study, we surveyed tumors that had spontaneously developed in Ink4a/Arf-deficient mice with an inbred FVB/NJ genetic background. We found that an Ink4a/Arf-deficiency exerted more severe effects on the induction of hematopoietic malignancies in mice with an inbred FVB/NJ genetic background than in mice with a mixed genetic background. We also provided the evidence that this prevalence of hematopoietic malignancies in Ink4a/Arf deficient mice is associated with the upregulated expressions of Stat5 and its transcriptional target, Bcl-x(L), both of which are involved in the regulation of hematopoiesis. These results suggest a possible implication of the Ink4a/Arf locus in the control of hematopoietic pathways by negatively regulating the Stat5 signalling pathways. PMID- 15888329 TI - Age-related alteration in hepatic acyl-CoA: cholesterol acyltransferase and its relation to LDL receptor and MAPK. AB - The aim of this study was to evaluate changes in the regulation of lipid metabolism and mitogen-activated protein kinases (MAPK) in the liver of C57BL/6 mice as they age. This was done by assessing the status of total cholesterol content and its enzyme, acyl-CoA: cholesterol acyltransferase (ACAT), in liver microsomal preparations and the low-density lipoprotein receptor (LDLr) mRNA expression in the livers of 4-24-month-old C57B/6 mice, without exogenous cholesterol feeding. With aging, there was an increase in cholesterol content and ACAT activity in liver microsomes. Northern blot analysis and real-time quantitative polymerase chain reaction data showed that ACAT-2 mRNA increased with age as well. LDLr expression decreased significantly in an age-dependent manner. In addition, we studied the basal and activated forms of MAPK, e.g. extracellular regulatory kinase (ERK-1/2), c-jun NH2-terminal kinase (JNK-1/2) and p38 MAPK. During aging, there was a considerable decrease in phosphorylated ERK-1/2 level while JNK-1/2 and p38 MAPK levels increased with age. Our studies showed an altered LDLr expression and altered phosphorylated MAPK in the liver of C57BL/6 mice during aging. These alterations might contribute to the development of atherosclerosis, hypercholesterolemia and other cholesterol-related conditions. PMID- 15888330 TI - A new mutation in the timing of autogamy in Paramecium tetraurelia. AB - We have isolated a new type of Paramecium tetraurelia mutant, named rie-2, that has a long immaturity period until autogamy. We previously isolated such an autogamy mutant, designated rie-1. These two mutants had some additional common features such as dependence of the occurrence of autogamy on the temperature, involvement of a single recessive gene, lower fission rate and shorter clonal life span. However, rie-2 was considered a new type mutant distinguishable from rie-1 because of their different natures of temperature sensitivity. First, the temperature at which they resembled the wild-type phenotype was low (19 degrees C) in rie-2, although it was high (32 degrees C) in rie-1. Second, the clonal life span of rie-2 at 25 degrees C was similar to that of the wild-type, but it was extremely shorter at 32 degrees C than at 25 degrees C, although it was similarly shorter at both temperatures in rie-1. Third, the difference of the fission rate between mutant and wild-type was greater at 32 degrees C than at 25 degrees C in rie-2, although it was similar at both temperatures in rie-1. This report shows that a gene mutation to elongate the period until sexual maturation does not necessarily assure the long life span. PMID- 15888331 TI - Proteasomal activity in brain differs between species and brain regions and changes with age. AB - Age-related increase in protein oxidation in brain coupled to an impairment of proteasomal activity may underline neuronal loss but differences in susceptibility between species and brain regions remain unexplained. We now investigate differences in proteasomal activity, measured as chymotrypsin-, trypsin- and peptidylglutamyl-like hydrolysing activities between brain regions in rats, mice and common marmosets. In aged rats and mice, proteasomal activity was decreased in the cortex, striatum, cerebellum, globus pallidus and substantia nigra overall when compared to young animals. However, in the aged brain only chymotrypsin-like activity was decreased in the cortex and the globus pallidus while only trypsin-like activity was reduced in the cerebellum. In contrast, in the striatum, both chymotrypsin-like and trypsin-like activities were reduced and in the substantia nigra, all the three catalytic activities of proteasome were significantly impaired. Chymotrypsin-like and trypsin-like activities were significantly higher in all the brain regions of marmosets compared to those of mice and rats. Peptidylglutamyl-like activity was only significantly higher in the cerebellum and striatum of marmoset compared to rodents. The data suggest that there is higher proteasome activity in common marmoset brain compared to rat and mouse and that the basal ganglia are more prone to an age-related decrease in proteasomal activity. This may explain the involvement of altered ubiquitin proteasome system activity in Parkinson's disease and the relationship to ageing. PMID- 15888332 TI - Natural ageing in the rat liver correlates with progressive stabilisation of DNA nuclear matrix interactions and withdrawal of genes from the nuclear substructure. AB - In the interphase nucleus, the DNA of higher eukaryotes is organised in supercoiled loops anchored to a nuclear matrix (NM). Replication, transcription and splicing seem to occur at macromolecular complexes organised upon the NM. Thus, the topological relationship between genes located in the loops and the NM appears to be very important for nuclear physiology. Here, we report that natural ageing in the rat liver correlates with a progressive strengthening of the NM framework and the stabilisation of the DNA loop-NM interactions, as well as with a progressive increase in the relative distance of genes to the NM. Both phenomena correlate with the gradual loss of proliferating potential and progression towards terminal differentiation in the hepatocytes, suggesting that wholesale modifications in the topological relationships within the cell nucleus are markers of tissue ageing and senescence, at least in the mammalian liver. We discuss the possible functional implications of such structural modifications that may underlie both terminal hepatocyte differentiation and their eventual replicative senescence. PMID- 15888333 TI - Energy expenditure of calorically restricted rats is higher than predicted from their altered body composition. AB - Debate exists over the impact of caloric restriction (CR) on the level of energy expenditure. At the whole animal level, CR decreases metabolic rates but in parallel body mass also declines. The question arises whether the reduction in metabolism is greater, smaller or not different from the expectation based on body mass change alone. Answers to this question depend on how metabolic rate is normalized and it has recently been suggested that this issue can only be resolved through detailed morphological investigation. Added to this issue is the problem of how appropriate the resting energy expenditure is to characterize metabolic events relating to aging phenomena. We measured the daily energy demands of young and old rats under ad libitum (AD) food intake or 40% CR, using the doubly labeled water (DLW) method and made detailed morphological examination of individuals, including 21 different body components. Whole body energy demands of CR rats were lower than AD rats, but the extent of this difference was much less than expected from the degree of caloric restriction, consistent with other studies using the DLW method on CR animals. Using multiple regression and multivariate data reduction methods we built two empirical predictive models of the association between daily energy demands and body composition using the ad lib animals. We then predicted the expected energy expenditures of the CR animals based on their altered morphology and compared these predictions to the observed daily energy demands. Independent of how we constructed the prediction, young and old rats under CR expended 30 and 50% more energy, respectively, than the prediction from their altered body composition. This effect is consistent with recent intra-specific observations of positive associations between energy metabolism and lifespan and theoretical ideas about mechanisms underpinning the relationship between oxygen consumption and reactive oxygen species production in mitochondria. PMID- 15888334 TI - Maximum longevities of chemically protected and non-protected fishes, reptiles, and amphibians support evolutionary hypotheses of aging. AB - Evolutionary hypotheses of aging predict that species with low rates of mortality from extrinsic sources, such as predation, should senesce more slowly and have longer maximum life spans than related species with higher rates of extrinsic mortality. We tested this prediction by synthesizing information on maximum body lengths and life spans in captivity of 1193 species of chemically protected (venomous or poisonous) and non-chemically protected fishes, snakes, caudatans (salamanders and newts), and anurans (frogs and toads). In every phylogenetic group maximum longevity was positively correlated with body size and, when size was controlled for statistically, chemically protected species and genera usually had longer maximum life spans than non-protected species. These results reemphasize the importance of life history traits, particularly protection from predation, in the evolution of senescence. PMID- 15888335 TI - Programming of hepatic antioxidant capacity and oxidative injury in the ageing rat. AB - Exposure to undernutrition in fetal life increases the susceptibility of humans and animals to major disease states in adult life. Studies of rodents exposed to protein restriction during fetal life indicate that lifespan is reduced, a finding that is the opposite of the well-documented improvement in longevity noted with post-weaning caloric restriction. The maternal low-protein (MLP) model of rat pregnancy therefore provides a suitable vehicle to consider how nutrition in fetal life may programme mechanisms of ageing. Pregnant rats were fed control (n = 7) or MLP diet (n = 7) throughout pregnancy. At birth, increased oxidation of hepatic proteins was noted, and this appeared related to activity of glutathione reductase. Older offspring were studied at 4, 16, 30 and 44 weeks of age and analyses revealed that MLP exposure in utero produced sex-specific differences in oxidant:antioxidant balance. In male offspring, MLP increased protein carbonyl concentrations in the liver (P = 0.013) and increased glutathione peroxidase activity (P = 0.018). These programmed changes were absent in the female offspring. The data support the hypothesis that reduced lifespan in rats subject to prenatal protein restriction is a consequence of enhanced oxidative processes promoting apoptosis and loss of tissue function. Mechanisms of ageing appear to be subject to nutritional programming during early development. PMID- 15888336 TI - Morpho-dynamic changes of mitochondria during ageing of human endothelial cells. AB - Mitochondrial morphology is regulated in many cultured eukaryotic cells by fusion and fission of mitochondria. A tightly controlled balance between fission and fusion events is required to ensure normal mitochondrial and cellular functions. During ageing, mitochondria are undergoing significant changes on the functional and morphological level. The effect of ageing on fusion and fission of mitochondria and consequences of altered fission and fusion activity are still unknown although theoretical models on ageing consider the significance of these processes. Human umbilical vein endothelial cells (HUVECs) have been established as a cell culture model to follow mitochondrial activity and dysfunction during the ageing process. Mitochondria of old and postmitotic HUVECs showed distinct alterations in overall morphology and fine structure, and furthermore, loss of mitochondrial membrane potential. In parallel, a decrease of intact mitochondrial DNA (mtDNA) was observed. Fission and fusion activity of mitochondria were quantified in living cells. Mitochondria of old HUVECs showed a significant and equal decrease of both fusion and fission activity indicating that these processes are sensitive to ageing and could contribute to the accumulation of damaged mitochondria during ageing. PMID- 15888337 TI - Cholesteryl ester transfer protein (CETP) I405V polymorphism and longevity in Italian centenarians. AB - A common polymorphism (I405V) in exon 14 of the cholesteryl ester transfer protein (CETP) gene has been recently associated to healthy aging in Ashkenazi Jewish. In order to study this genetic effect in long-lived individuals with a different ethnicity, we analyzed the allele and genotype distributions of the CETP polymorphism a sample of Italian centenarians. Our result does not confirm the association between the I405V CETP variation and the healthy aging phenotype described in the Ashkenazi Jewish population and suggests that other gene environment interactions contribute to longevity. PMID- 15888338 TI - IgA levels are predictors of mortality in Finnish nonagenarians. AB - It has been demonstrated that in obviously healthy, very old people increased levels of inflammatory markers as well as some defects in T lymphocyte populations are strong predictors of mortality. Very little is known about the role of possible functional defects in antibody formation. To examine this, we now measured IgM, IgG and IgA concentrations in a cohort of 285 nonagenarians (67 males, 218 females). IgG and IgA levels were significantly higher than those of healthy middle-aged controls. The analyzed serum samples were taken at the age of 90-91 years. After 4 years, 20 males and 94 females had survived. To analyze the role in predicting mortality, the immunoglobulin data (as well as the measured CRP and IL-6 concentrations) were stratified according to this survival data. IgA levels (and CRP and IL-6 levels) were clearly higher in the nonsurvivors than in the survivors. These data imply that elevated serum IgA level, i.e. indicator of intestinal inflammation and/or defect in mucosal defence, is a strong mortality predicting factor. PMID- 15888339 TI - A new formalization of a meta-game using the lambda calculus. AB - This paper presents a new game system formalism. The system describes both strategies and a game master (who computes scores in a given game system) in terms of lambda-calculus. This formalism revisits the prisoner's dilemma game, to discuss how meta-strategies emerge in this classical game, even without repetition. We have also examined the evolution of meta-strategies in lambda formalism. PMID- 15888340 TI - "Live memory" of the cell, the other hereditary memory of living systems. AB - We propose to designate by the term "live memory" of the cell, the cytoplasmic memory. This phenomenon consists of non-genetic memory, but nevertheless includes transmission function, which may be "hereditary" via the ovum, from mother cell to daughter cell, or simply within the same cell from instant t to instant t+1. To understand this notion of "live memory", its role and interactions with DNA must be resituated; indeed, operational information belongs as much to the cell body and to its cytoplasmic regulatory protein components and other endogenous or exogenous ligands as it does to the DNA database. We will see in Section 2, using examples from recent experiments in biology, the principal roles of "live memory" in relation to the four aspects of cellular identity, memory of form, hereditary transmission and also working memory. PMID- 15888341 TI - Indispensable role of quantum theory in the brain dynamics. AB - Recently, Tegmark pointed out that the superposition of ion states involved in the superposition of firing and resting states of a neuron quickly decohere. It undoubtedly indicates that neural networks cannot work as quantum computers, or computers taking advantage of coherent states. Does it also mean that the brain can be modeled as a neural network obeying classical physics? Here we show that it does not mean that the brain can be modeled as a neural network obeying classical physics. A brand new perspective in research of neural networks from quantum theoretical aspect is presented. PMID- 15888342 TI - A hybrid systems framework for cellular processes. AB - With the availability of technologies that allow us to obtain stimulus-response time series data for modeling and system identification, there is going to be an increasing need for conceptual frameworks in which to formulate and test hypotheses about intra- and inter-cellular dynamics, in general and not just dependent on a particular cell line, cell type, organism, or technology. While the semantics can be quite different, biologists and systems scientists use in many cases a similar language (notion of feedback, regulation, etc.). A more abstract system-theoretic framework for signals, systems, and control could provide the biologist with an interface between the domains. Apart from recent examples to identify functional elements and describing them in engineering terms, there have been various more abstract developments to describe dynamics at the cell level in the past. This includes Rosen's (M,R)-systems. This paper presents an abstract and general compact mathematical framework of intracellular dynamics, regulation and regime switching inspired by (M,R)-theory and based on hybrid automata. PMID- 15888343 TI - UML as a cell and biochemistry modeling language. AB - The systems biology community is building increasingly complex models and simulations of cells and other biological entities, and are beginning to look at alternatives to traditional representations such as those provided by ordinary differential equations (ODE). The lessons learned over the years by the software development community in designing and building increasingly complex telecommunication and other commercial real-time reactive systems, can be advantageously applied to the problems of modeling in the biology domain. Making use of the object-oriented (OO) paradigm, the unified modeling language (UML) and Real-Time Object-Oriented Modeling (ROOM) visual formalisms, and the Rational Rose RealTime (RRT) visual modeling tool, we describe a multi-step process we have used to construct top-down models of cells and cell aggregates. The simple example model described in this paper includes membranes with lipid bilayers, multiple compartments including a variable number of mitochondria, substrate molecules, enzymes with reaction rules, and metabolic pathways. We demonstrate the relevance of abstraction, reuse, objects, classes, component and inheritance hierarchies, multiplicity, visual modeling, and other current software development best practices. We show how it is possible to start with a direct diagrammatic representation of a biological structure such as a cell, using terminology familiar to biologists, and by following a process of gradually adding more and more detail, arrive at a system with structure and behavior of arbitrary complexity that can run and be observed on a computer. We discuss our CellAK (Cell Assembly Kit) approach in terms of features found in SBML, CellML, E CELL, Gepasi, Jarnac, StochSim, Virtual Cell, and membrane computing systems. PMID- 15888345 TI - Transgenic mouse models for the prevention of breast cancer. AB - Breast cancer prevention research has made remarkable progress in the past decade. Much of this progress has come from clinical trials. However, in the future to test the many promising agents that are now available, pre-clinical models of breast cancer are needed. Such models are now available. Useful models include rat and mouse models, particularly, the genetically engineered mice (GEM). Many transgenic mouse models have been generated by manipulating growth factors and their receptors, cell cycle regulators, signal transduction pathways, cellular differentiation, oncogenes and tumor suppressor genes. The transgenes are induced to express in the mouse mammary glands under the control of various transgenic promoters, which have respective characteristics in expression pattern and other biological attributes. These models are providing invaluable insight on the molecular mechanisms of breast tumorigenesis. In this review, we discuss the relative relevance of the most commonly used transgenic mouse models for breast cancer prevention studies, and provide examples of how these transgenic models can be used to conduct cancer prevention research. Due to the multi-factor, multi step nature of breast cancer, many factors should be incorporated into a valid prevention study. However, many barriers to progress must be overcome, including access to and availability of new cancer preventive drugs, and difficulties in conducting studies of combinations of preventive agents. PMID- 15888344 TI - Mouse models expressing human carcinoembryonic antigen (CEA) as a transgene: evaluation of CEA-based cancer vaccines. AB - In recent years, investigators have carried out several studies designed to evaluate whether human tumor-associated antigens might be exploited as targets for active specific immunotherapy, specifically human cancer vaccines. Not too long ago such an approach would have been met with considerable skepticism because the immune system was believed to be a rigid discriminator between self and non-self which, in turn, protected the host from a variety of pathogens. That viewpoint has been challenged in recent years by a series of studies indicating that antigenic determinants of self have not induced absolute host immune tolerance. Moreover, under specific conditions that evoke danger signals, peptides from self-antigen can be processed by the antigen-presenting cellular machinery, loaded onto the major histocompatibility antigen groove to serve as targets for immune intervention. Those findings provide the rationale to investigate a wide range of tumor-associated antigens, including differentiation antigens, oncogenes, and tumor suppressor genes as possible immune-based targets. One of those tumor-associated antigens is the carcinoembryonic antigen (CEA). Described almost 40 years ago, CEA is a M(r) 180-200,000 oncofetal antigen that is one of the more widely studied human tumor-associated antigens. This review will provide: (i) a brief overview of the CEA gene family, (ii) a summary of early preclinical findings on overcoming immune tolerance to CEA, and (iii) the rationale to develop mouse models which spontaneously develop gastrointestinal tumors and express the CEA transgene. Those models have been used extensively in the study of overcoming host immune tolerance to CEA, a self, tumor-associated antigen, and the experimental findings have served as the rationale for the design of early clinical trials to evaluate CEA-based cancer vaccines. PMID- 15888346 TI - Novel p38 MAP kinase inhibitor R-130823 suppresses IL-6, IL-8 and MMP-13 production in spheroid culture of human synovial sarcoma cell line SW 982. AB - Synovial hyperplasia is a hallmark of rheumatoid arthritis (RA) and is regarded as a major destructive element of articular bone and cartilage. This pathological process is accompanied by the production of proinflammatory cytokines, prostaglandin E(2) (PGE(2)), and matrix metalloproteinases (MMPs) in synoviocytes. We studied the spontaneous production of these substances in RA synoviocytes in spheroid culture. Synovial sarcoma cell line SW 982 formed a single spheroid in non-adherent culture plates. It produced interleukin (IL) 1beta, IL-6, IL-8, PGE(2), MMP-2 and MMP-13. Neither the addition of integrin antagonizing oligopeptide (GRGDSP) nor that of vitronectin receptor inhibitor SB 265123 to the culture inhibited any production. Phosphorylation of p38 mitogen activated protein (MAP) kinase was observed during the culture. A novel p38 MAP kinase inhibitor, R-130823, inhibited the release of IL-6, IL-8 and MMP-13 in a concentration-dependent manner, but not that of IL-1beta or MMP-2. Real-time RT PCR analysis demonstrated that IL-6, IL-8 and MMP-13 were inhibited at the transcriptional level. R-130823 did not affect the production of PGE(2) in spheroid culture, while the addition of R-130823 suppressed IL-1beta-induced PGE(2) synthesis in monolayer culture of SW 982 cells. The results suggest that spheroid culture induced proinflammatory factors and MMPs in signaling pathways both dependent and independent of p38 MAP kinase. PMID- 15888347 TI - Effect of recombinant human BMP-2 on the repair of cricoid cartilage defects in young and adult rabbits: a comparative study. AB - OBJECTIVE: The study evaluated the possible differences in the repair of cricoid cartilage defects treated with recombinant human BMP-2 in young and adult rabbits. METHODS: A cricoid defect rabbit model was used. Thirty rabbits were randomly divided into eight groups. Two groups of young rabbits and two groups of adult rabbits were treated with rhBMP-2 delivered on an absorbable collagen sponge, while the other two groups of young rabbits and two groups of adult rabbits were used as controls. The rabbits were killed at 1 week or 4 weeks after surgery. A histomorphometric analysis and an evaluation of the expression of collagen types I, II, and X, and proliferating cell nuclear antigen as well as a study of distribution of calcified matrix, were performed. RESULTS: rhBMP-2 induced a marked chondrogenesis in both experimental age groups. However, in young rabbits the newly formed cartilage appeared more elongate, and the length of perichondrium involved was greater. The host cricoid cartilage of adult rabbits was calcified in large areas and displayed a strong matrix expression of collagen type X as well as collagen type I in the perichondrium, compared to the cricoid of young rabbits. In spite of these differences no immunohistochemical differences were found in the newly formed cartilage of both age groups treated with rhBMP-2. The cricoid cartilage defect was filled with new bone at 4 weeks in both age groups treated with rhBMP-2. New bone tissue had a well-defined trabecular structure. CONCLUSIONS: rhBMP-2 triggers appositional cartilage growth from the cricoid perichondrium of young rabbits more easily than from that of adult rabbits. The new bone induced by rhBMP-2 showed a similar immunohistochemical and morphological pattern in both age groups of rabbits. PMID- 15888348 TI - The Phanerochaete chrysosporium secretome: database predictions and initial mass spectrometry peptide identifications in cellulose-grown medium. AB - The white rot basidiomycete, Phanerochaete chrysosporium, employs an array of extracellular enzymes to completely degrade the major polymers of wood: cellulose, hemicellulose and lignin. Towards the identification of participating enzymes, 268 likely secreted proteins were predicted using SignalP and TargetP algorithms. To assess the reliability of secretome predictions and to evaluate the usefulness of the current database, we performed shotgun LC-MS/MS on cultures grown on standard cellulose-containing medium. A total of 182 unique peptide sequences were matched to 50 specific genes, of which 24 were among the secretome subset. Underscoring the rich genetic diversity of P. chrysosporium, identifications included 32 glycosyl hydrolases. Functionally interconnected enzyme groups were recognized. For example, the multiple endoglucanases and processive exocellobiohydrolases observed quite probably attack cellulose in a synergistic manner. In addition, a hemicellulolytic system included endoxylanases, alpha-galactosidase, acetyl xylan esterase, and alpha-l arabinofuranosidase. Glucose and cellobiose metabolism likely involves cellobiose dehydrogenase, glucose oxidase, and various inverting glycoside hydrolases, all perhaps enhanced by an epimerase. To evaluate the completeness of the current database, mass spectroscopy analysis was performed on a larger and more inclusive dataset containing all possible ORFs. This allowed identification of a previously undetected hypothetical protein and a putative acid phosphatase. The expression of several genes was supported by RT-PCR amplification of their cDNAs. PMID- 15888349 TI - Dynamic optimization of bioprocesses: efficient and robust numerical strategies. AB - The dynamic optimization (open loop optimal control) of non-linear bioprocesses is considered in this contribution. These processes can be described by sets of non-linear differential and algebraic equations (DAEs), usually subject to constraints in the state and control variables. A review of the available solution techniques for this class of problems is presented, highlighting the numerical difficulties arising from the non-linear, constrained and often discontinuous nature of these systems. In order to surmount these difficulties, we present several alternative stochastic and hybrid techniques based on the control vector parameterization (CVP) approach. The CVP approach is a direct method which transforms the original problem into a non-linear programming (NLP) problem, which must be solved by a suitable (efficient and robust) solver. In particular, a hybrid technique uses a first global optimization phase followed by a fast second phase based on a local deterministic method, so it can handle the nonconvexity of many of these NLPs. The efficiency and robustness of these techniques is illustrated by solving several challenging case studies regarding the optimal control of fed-batch bioreactors and other bioprocesses. In order to fairly evaluate their advantages, a careful and critical comparison with several other direct approaches is provided. The results indicate that the two-phase hybrid approach presents the best compromise between robustness and efficiency. PMID- 15888350 TI - An optimal code for patient identifiers. AB - How to distinguish 1 billion individuals by an identifier consisting of eight characters, allowing a reasonable amount of error detection or even error correction? Our solution of this problem is an optimal code over a 32-character alphabet that detects up to two errors and corrects one error as well as a transposition of two adjacent characters. The corresponding encoding and error checking algorithms are available for free; they are also embedded as components of the pseudonymisation service that is used in the TMF-the German telematics platform for health research networks. PMID- 15888351 TI - Apoptosis regulation and drug resistance in malignant pleural mesothelioma. PMID- 15888352 TI - Clinical outcome and technical aspects of 263 radial forearm free flaps used in reconstruction of the oral cavity. AB - BACKGROUND: The aim of this study was to report the applications, complications, and limitations of the radial forearm flap when used for reconstruction after excision of cancers in the mouth. PATIENTS AND METHODS: We reviewed records in our database from October 1987 to December 2002; a total of 505 patients had surgical defects of the head and neck reconstructed with a radial forearm flap: 258 patients had oral cancers, of whom 173 were men and 85 women, median age was 70 years. Five patients had two reconstructions (total 263), of which 247 were fasciocutaneous and 16 osseofasciocutaneous flaps. Mucosal squamous carcinoma accounted for 97% of the primary cancers. RESULTS: There were nine failures of the flap (3.4%) and four episodes of partial necrosis. Four patients (2%) died within 30 days of operation. Orocutaneous fistulas appeared in nine patients (3.4%), 10 patients had wound infections, and nine had haematomas. The nine failed flaps were replaced by a second free flap in one patient, a pectoralis major flap in three, a buccinator myomucosal flap in one, a skin graft in one, and the remaining three defects healed by secondary intention. There was one failure and one partial failure among the 16 osseofasciocutaneous flaps. PMID- 15888353 TI - The composite radial forearm free flap: an anatomical guide to harvesting the radius. AB - We made an anatomical study of 100 dried cadaveric radii to establish the dimensions of the maximum possible safe harvest of bone for the composite radial forearm flap and how this may be assessed during the operation. We found that the diameter of the radius from its medial to lateral border is consistently 1-3mm less than 40% of the measured minimum circumference of the radius. PMID- 15888354 TI - INTERESTING CASE: cordless phone aerial in maxillary sinus. PMID- 15888355 TI - Free vascularised iliac crest graft: an audit of 26 consecutive cases. AB - Over an 18 month period 26 vascularised myo-osseus, and one myo-osseus-cutaneous iliac crest flaps were used to reconstruct 18 mandibular and eight maxillary defects. During the operation one flap failed to reperfuse and was removed. Postoperatively three patients were returned to theatre for appraisal and reanastomoses, which in two cases involved thrombolysis. In all three salvage was deemed successful but in all there was partial muscle necrosis. There were 10 cases of anaesthesia of the skin supplied by the lateral cutaneous nerve of the thigh, three incisional hernias, one wound dehiscence in the thigh, two oronasal fistulas, one wound dehiscence in the neck, one infection by methicillin resistant Staphylococcus aureus in the thigh, and two long-term and one medium term disturbances of gait. Complications were graded as severe (4%), intermediate (27%), and minor (46%). PMID- 15888356 TI - Thromboelastographic study of the effect of manipulation of central veins on coagulability of venous blood. AB - AIM: To measure whole blood coagulability in vitro using thromboelastography before and after direct and indirect manipulation of the internal jugular and subclavian veins. METHODS: We studied 10 patients (American Society of Anaesthesiologists (ASA) grades I or II) with malignant disease of the head and neck who were having selective neck dissections. We excluded patients with diabetes, those who were being given any anticoagulant, those who were actively infected, and those with known coagulopathy. We took 3 ml samples of venous blood during insertion of a peripheral cannula, after insertion of a central venous cannula, and after dissection of the internal jugular vein. Thromboelastography was done on recalcified citrated samples within 1h of sampling. The variables r, k, alpha angle, and maximum amplitude were recorded at each point and the values before and after manipulation were compared using t-tests. RESULTS: The only change was seen in the variable r, which showed an insignificant increase in coagulability. PMID- 15888357 TI - Survey of the use of hyperbaric oxygen by maxillofacial oncologists in the UK. AB - Oral and maxillofacial surgeons often use hyperbaric oxygen (HBO). Our aim was to find out the referral pattern of these surgeons for HBO. We contacted oral and maxillofacial units in England, Wales, and Scotland and identified 125 consultants who are involved in the management of patients with cancers of the head and neck. We sent these surgeons a postal questionnaire and 91 (73%) replied. Eighty-five of these consultants (93%) saw patients with osteoradionecrosis and only five of these never referred patients for HBO. About half the respondents (57%) saw patients for the insertion of osseointegrated implants after radiotherapy to the jaw, and seven of these never referred patients for HBO. All the respondents saw patients who required mandibular molar extractions after radiotherapy and 30 (33%) never referred these patients for HBO. Most consultants were unaware of the method of delivery of HBO. This survey suggests that most surgeons consider HBO to be part of the management of osteoradionecrosis, but their knowledge about delivery is weak and protocols vary. PMID- 15888358 TI - Orofacial tumours in suburban Nigerian children and adolescents. AB - We report the type and distribution of orofacial tumours in south-western Nigerian children and adolescents. The 512 records of patients with oral and maxillofacial tumours in the Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, from 1991 to 2001, were searched and the 146 records (28%) of patients aged 19 years and less were reviewed. Their mean age was 10 years (female:male ratio 1:1.4). Of the 146 tumours 74 (51%) were malignant and 72 (49%) were benign. Of the latter 31 were from soft tissue and 41 were from the jaw (20 odontogenic and 21 non-odontogenic). The most common benign soft tissue and jaw tumours were gingival epulis and ameloblastoma respectively. Of the malignant tumours 67 were lymphomas, 5 sarcomas and 2 carcinomas. PMID- 15888359 TI - Objective evaluation of iatrogenic lingual nerve injuries using the jaw-opening reflex. AB - The extent of reflex inhibition of masseteric electromyographic activity, after an electrical stimulus applied to lingual mucosa, was used as a test of the ability of the lingual nerve to conduct nerve impulses and this was compared with the results of standard clinical tests. Two groups of subjects were assessed: healthy subjects (n=10) and patients with lingual nerve injuries (n=17). The patients were tested 8-9 weeks after their injury and retested 6 months later when they were retrospectively allocated to either a temporary injury or a permanent injury group. The group measure of reflex inhibition after stimulation of the tongue on the opposite side to the injury was no different from the same measure in controls, whereas two-point discrimination did differ. Group measures of inhibition and of subjective function after stimulation on the side of the injury were significantly different from controls whereas light touch and two point discrimination were not. There was good agreement between quantified masseteric inhibition and subjective function, but it was not possible at 8-9 weeks after the injury to differentiate between those that would recover and those that would be permanent. PMID- 15888360 TI - New method for the objective evaluation of injury to the lingual nerve after operation on third molars. AB - Existing tests of function of the lingual nerve are either subjective or, when they elicit the jaw-opening reflex, are dependent on the cooperation of the subject. We report a study in 12 healthy volunteers and 12 patients with iatrogenic injury to the lingual nerve. A bite block (containing stimulating electrodes) was held between the teeth and the tongue was held on to the electrodes by suction. When the lingual nerve was intact, an electrical stimulus elicited brief inhibition of masseteric electromyographic activity. Local analgesia and iatrogenic injury to the lingual nerve altered nerve conduction and caused a reduction in reflex inhibition. Two methods, compatible with limited numbers of applications of the stimulus, were used to quantify responses. One used an indirect measurement of intervals between action potentials of muscle and the other used a measurement of rectified signals falling below the mean amplitude before and after the stimulus. Both methods gave values that correlated with subjective sensations. The first gave an estimate of the probability of defining major malfunction of the nerve objectively; the second gave a linear measurement that allowed recovery of the nerve to be followed. PMID- 15888361 TI - Ultrasound-guided basket retrieval of salivary stones: a new technique. AB - We describe the successful use of ultrasound-guided retrieval of a submandibular stone with a basket in two patients. PMID- 15888362 TI - Angiolymphoid hyperplasia with eosinophilia masquerading as Kimura disease. AB - Angiolymphoid hyperplasia with eosinophilia is a rare benign vasoproliferative disease of unknown cause, which can present as tumour-like nodules in the head and neck. We treated a patient who showed some of the features of Kimura disease (KD), which is an important differential diagnosis, because of the latter's association with renal disease. PMID- 15888363 TI - Advice given in community pharmacies to patients with possible oral carcinoma. AB - We compared advice given by community pharmacy staff in Worcestershire to a proxy consulter on behalf of a patient with a longstanding oral ulcer, presumed to be an oral cancer. If patients seek advice in this way the quality of the advice given is dependent on whether the respondent is a pharmacist or a community pharmacy assistant. PMID- 15888364 TI - Improved position for the flexible laryngeal mask. PMID- 15888365 TI - Minimally invasive open reduction of a displaced condylar fracture in a child. AB - We treated successfully a severely displaced condylar fracture in a child 9 years of age by endoscopically assisted open operation. PMID- 15888366 TI - Osteomyelitis of the mandible during pregnancy. AB - A 21-year-old woman who was 9 weeks pregnant presented with osteomyelitis of the mandible that resolved spontaneously after delivery. To our knowledge a link between osteomyelitis of the jaws and pregnancy has not been reported previously. PMID- 15888367 TI - View of the epiglottis during examination of the oral cavity. PMID- 15888368 TI - Sentiments expressed both at the recent BAOMS Medical Students Group/Junior Trainees Conference and in the Presidential Newsletter of October 2003. PMID- 15888369 TI - Re: Gibbones AJ, Moss C, Robertson JM, Consley R. Fast tracking of referrals for orthodontic oral surgery. Br J Oral Maxillofac Surg 2004;42:277-8. PMID- 15888370 TI - Dental anaesthesia in a patient with Noonan syndrome. PMID- 15888371 TI - Trainee conference debate on the effect of fellowships on maxillofacial training in the United Kingdom. PMID- 15888372 TI - Re: Martin TJM, Sharp I, Oral mucosal pigmentation secondary to treatment with mepacrine, with sparing of the denture bearing area (Br J Oral Maxillofac Surg 2004;42:351-3). PMID- 15888373 TI - Classification and treatment of grossly oblique mandibular fractures. PMID- 15888374 TI - Re: Kurian A, Ward-Booth P, Blood transfusion and orthognathic surgery--a thing of the past? PMID- 15888375 TI - Morphological alteration in oro-facial CGRP containing motoneurons due to congenital thyroid hypofunction. AB - Under congenital thyroid hypofunction, the oro-facial large and small calcitonin gene-related peptide (CGRP) immunoreactive motoneurons were classified into strong, moderate, weak and negative intensity in offspring weaned rats. While 50% of neurons in the trigeminal motor nucleus (Mo5) were of the large type, this value dropped to 30% in hypothyroid pups. Hypothyroid trigeminal accessory nucleus (Mo5-AC) contained 10% large motoneurons versus about 45% in normal pups. Normal facial nucleus (Mo7) had 20% large motoneurons in contrast with 10% in hypothyroid pups. These values are significant in comparison with the normal pattern of oro-facial CGRP positive immunoreactive motoneurons as well as those devoid of immunostaining. PMID- 15888376 TI - Response of brain amino acid metabolism to ketosis. AB - Our objective was to study brain amino acid metabolism in response to ketosis. The underlying hypothesis is that ketosis is associated with a fundamental change of brain amino acid handling and that this alteration is a factor in the anti epileptic effect of the ketogenic diet. Specifically, we hypothesize that brain converts ketone bodies to acetyl-CoA and that this results in increased flux through the citrate synthetase reaction. As a result, oxaloacetate is consumed and is less available to the aspartate aminotransferase reaction; therefore, less glutamate is converted to aspartate and relatively more glutamate becomes available to the glutamine synthetase and glutamate decarboxylase reactions. We found in a mouse model of ketosis that the concentration of forebrain aspartate was diminished but the concentration of acetyl-CoA was increased. Studies of the incorporation of 13C into glutamate and glutamine with either [1-(13)C]glucose or [2-(13)C]acetate as precursor showed that ketotic brain metabolized relatively less glucose and relatively more acetate. When the ketotic mice were administered both acetate and a nitrogen donor, such as alanine or leucine, they manifested an increased forebrain concentration of glutamine and GABA. These findings supported the hypothesis that in ketosis there is greater production of acetyl-CoA and a consequent alteration in the equilibrium of the aspartate aminotransferase reaction that results in diminished aspartate production and potentially enhanced synthesis of glutamine and GABA. PMID- 15888377 TI - Pdx1-related homeodomain transcription factors are distinctly expressed in mouse adult pancreatic islets. AB - Complex gene networks are responsible for the proper operation of the endocrine pancreas. A central member of such networks, the homeodomain transcription factor Pdx1, belongs to the ParaHox gene cluster, an array of Hox-like homeobox genes. With a combination of mRNA in situ hybridisation and immunodetection, we have found that the rest of ParaHox cluster genes, Cdx1, Cdx2/3, and Cdx4, and Gsh1 and Gsh2, are all expressed in specific islet cell types of the endocrine pancreas. To our knowledge, this is the first report that locates ParaHox genes other than Pdx1 and Cdx2/3 in a place as to be involved in the pancreatic transcriptional regulatory networks, potentially regulating glucagon-insulin homeostasis. PMID- 15888378 TI - Cytotoxicity of plants from Malaysia and Thailand used traditionally to treat cancer. AB - The SRB cytotoxicity assay was used to screen extracts and isolated constituents of some traditional medicinal plants from Malaysia and Thailand against two human cancer cell lines, COR L23 lung cancer cell line and MCF7 breast cancer cell line and the non-cancer MCF5 cell line. Five out of the seven species tested, i.e. Thai Alpinia galanga, Alpinia officinarum, Cayratia japonica, Physalis minima, Tabernaemontana divaricata, exhibited interesting cytotoxicity activity and this is the first report of cytotoxicity from any Cayratia species. Following bioassay guided fractionation, 1'-acetoxychavicol acetate (48h exposure against COR L23 cells, IC(50) 7.8 microM against MCF7 cells, IC(50) 23.9 microM) was isolated as the major cytotoxic component of the Alpinia species, physalin F as the major cytotoxic component of Physalis minima (48 h exposure against COR L23 cells IC(50) 0.4 microM against MCF7 cells, IC(50) 0.59 microM). The Malaysian Alpinia galanga showed weak activity compared with the Thai sample and this was shown to be due to the relatively high amounts of 1'-acetoxychavicol acetate present in the Thai sample. PMID- 15888379 TI - Blood flow: a key regulatory component of corpus luteum function in the cow. AB - Prostaglandin F2alpha (PGF2alpha) is the primary luteolysin in the cow. During the early luteal phase, the corpus luteum (CL) is resistant to the luteolytic effect of PGF2alpha. Once mature, the CL becomes responsive to PGF2alpha and undergoes luteal regression. These actions of PGF2alpha coincide with changes in luteal blood flow (BF): PGF2alpha has no effect on BF in the early CL, but acutely increases BF in the peripheral vasculature of the mature CL within 30 min of PGF2alpha injection. During spontaneous luteolysis, luteal BF increases on Days 17-18 of the estrous cycle, prior to any decrease in plasma progesterone (P). The increase in luteal BF is synchronous with an increase in plasma PGFM levels, suggesting that pulsatile release of PGF2alpha from uterus stimulates the increase in luteal BF. Serial biopsies of these CL showed that mRNA expression for endothelial nitric oxide synthase (eNOS) together with endothelin-1 (ET-1) and angiotensin converting enzyme (ACE) increases on Days 17-18 when the luteal BF is elevated. On Day 19 when plasma P level firstly decreases, eNOS mRNA returns to the basal level whereas ET-1 and ACE mRNA remains elevated. Cyclooxygenase-2 (COX-2) mRNA expression increases on Day 19. In support of these data, an in vivo microdialysis study revealed that luteal ET-1 and angiotensin II (Ang II) secretion increases and precedes PGF2alpha secretion during spontaneous luteolysis. In conclusion, we show for the first time that an acute increase of BF occurs in the peripheral vasculature of the mature CL together with increases in eNOS expression and ET-1 and Ang II secretion in the CL during the early stages of luteolysis in the cow. We propose that the increase in luteal BF may be induced by NO from large arterioles surrounding the CL, and simultaneously uterine or exogenous PGF2alpha directly increases ET-1 and Ang II secretion from endothelial cells of microcapillary vessels within the CL, thereby suppressing P secretion by luteal cells. Taken together, our results indicate that an acute increase in luteal BF occurs as a first step of luteolysis in response to PGF2alpha. Therefore, local BF plays a key role to initiate luteal regression in the cow. PMID- 15888380 TI - Exon-3 polymorphism of CTLA-4 gene in Turkish patients with vitiligo. PMID- 15888381 TI - Expression of the gonadal p450 aromatase gene of Xenopus and characterization of the 5'-flanking region of the aromatase gene. AB - Investigation by RQ-RT-PCR revealed that transcription of the p450 aromatase gene is activated at stage 50, when sex determination of the female begins, and that aromatase gene expression is also activated by exogenously administrated estradiol. In order to determine the molecular basis underlying the specific activation of aromatase gene expression during sex differentiation and in response to exogenous estradiol, we isolated the 5'-flanking fragment of the gene and characterized the promoter sequence. We demonstrated binding sequences to a specific trans-activating factor upstream of the p450 aromatase promoter II, the cAMP response element binding protein/activating transcription factor family, and steroidogenic factor-1. An estrogen response element half-site sequence that recognize an estrogen receptors, was also found. PMID- 15888382 TI - Potentiality of combined hepatocellular and intrahepatic cholangiocellular carcinoma originating from a hepatic precursor cell: Immunohistochemical evidence. AB - Hypothesizing that combined hepatocellular and intrahepatic cholangiocellular carcinoma originates from hepatic stem cells, we conducted a series of immunohistochemical studies using hepatic stem cell markers to better understand the origin of the tumor. Specially, we investigated 15 combined tumors confirmed to be immunoreactive to anti-hepatocyte paraffin 1, and anti-cytokeratin 7 and 19 antibodies. Macroscopic investigation revealed cancer cells morphologically intermediate between neoplastic hepatocytes and cholangiocytes (combined tumor with intermediate morphology) in 11 of 15 tumors. The remaining four tumors lacking these cells were considered ordinary combined tumors. We used 20 hepatocellular carcinomas and 10 cholangiocarcinomas as controls. Immunohistochemical analysis was performed using stem cell markers (c-kit and CD34). Of the 15 combined tumors, 8 stained for c-kit (2 of the 4 ordinary tumors and 6 of the 11 intermediate morphologic tumors). Both of the tumor cell types in both of the ordinary combined tumors stained for c-kit. The c-kit staining was present in cells of all three morphologic types, including the intermediate cells, in all six of the intermediate combined tumors. None of the cells in the combined tumors showed immunoreactivity for CD34. None of the 30 control tumors expressed CD 34 or c-kit. The present study suggests that combined hepatocellular and intrahepatic cholangiocellular carcinomas originate not from a hepatic stem cell, but from a hepatic precursor cell, such as the canal of Hering cell. Alternatively, these tumors might show up-regulation of c-kit in relation to angiogenesis. PMID- 15888383 TI - A practical overview of aromatase inhibitors in postmenopausal women with hormone receptor-positive breast cancer. AB - Tamoxifen has been the standard adjuvant therapy for patients with breast cancer for the last several decades. Several recently completed adjuvant trials using aromatase inhibitors have shown the superiority of these agents over Tamoxifen or placebo, when used in postmenopausal women. The results from these trials present a challenging situation for practicing oncologists with regards to the choice, duration, sequence of therapy, follow up and side effects of aromatase inhibitors. We have discussed various management issues from a practical angle for oncologists to effectively use these agents, with an evidence-based approach. PMID- 15888384 TI - [From intellectual property to commercial property]. PMID- 15888386 TI - [Histological classification of human gliomas: state of art and controversies]. AB - The histological classification of human gliomas remains in 2005 a challenge. The aim is to define the histological type of glioma (astrocytic, oligodendrocytic or mixed) and the grade in order to classify the patients and give them an accurate treatment. Although the standard remains the WHO classification, this classification suffered from lack of reproducibility among pathologists. In particular this classification does not take into account the intrinsic morphological heterogeneity of infiltrative gliomas and does not discriminate the tumour cells from the residual brain parenchyma. According to the WHO classification, infiltrative gliomas encompass astrocytic gliomas (diffuse astrocytomas grade II, anaplastic astrocytomas grade III and glioblastomas grade IV), oligodendroglial tumours (oligodendrogliomas grade II, anaplastic oligodendrogliomas grade III) and mixed gliomas (oligoastrocytomas grade II and anaplastic oligoastrocytomas grade III). Circumscribed gliomas mainly corresponds to pilocytic astrocytomas (grade I). In contrast, the Sainte Anne classification takes into account the macroscopic informations provided by imaging techniques and the tumour growth patterns. Three distinct tumour growth patterns may be seen in gliomas, type I: tumor tissue only, type II: tumour tissue and isolated tumor cells permeating the brain parenchyma (ITC) and type III: ITCs only and no tumor tissue. According to the Sainte Anne classification, gliomas are divided into astrocytic gliomas (pilocytic astrocytomas, structure type I, glioblastomas structure type II) and oligodendrogliomas and mixed oligoastrocytomas (grade A: lack of contrast enhancement and lack of endothelial hyperplasia, structure type III; and grade B: contrast enhancement or endothelial hyperplasia, structure type II and III). In the future the glioma classification has to be unique and should take into account clinical data, neuroradiological and histological features and results of molecular biology. PMID- 15888387 TI - [Towards a molecular classification of gliomas]. AB - Several molecular genetic alterations have been characterized in gliomas in the past years. Molecular profiles have been associated with specific histologic and prognostic tumor subgroups, contributing to improve the classification of gliomas. At least two alternative molecular pathways have been suggested in the astrocytoma progression involving TP53 inactivation (secondary glioblastomas) and EGFR amplification (de novo glioblastomas) respectively. Oligodendroglial tumors have demonstrated recurrent combined loss of chromosome 1p/19q, which represent a favorable prognosis marker and probably a predictor of a good chemosensitivity of the tumor. This review discusses recent molecular advances and clinical implications with special focus on oligodendroglial tumors. PMID- 15888388 TI - [Role of adrenomedullin in glioblastomas growth]. AB - Glioblastoma multiforme is the most malignant of the primary brain tumors and is almost always fatal. The treatment strategies for this disease have not changed appreciably for many years and most are based on a limited understanding of the biology of the disease. Growth factors are potential targets for therapeutic strategies because they are essential for tumor growth and progression. Adrenomedullin (AM) is a multifunctional regulatory peptide with mitogenic and angiogenic capabilities among others. Real-time quantitative reverse transcriptase-polymerase chain reaction analysis showed that AM mRNA was correlated to the tumor type and grade, with high expression in all glioblastomas analysed, whereas a low expression was found in anaplastic astrocytomas and barely detectable levels in low-grade astrocytomas and oligodendriogliomas. The correlation of AM expression to the grade of glioma support the hypothesis that AM may participate in the progression of gliomas. We demonstrate that AM may function as an autocrine/paracrine growth factor for glioblastoma cells. The data demonstrated that the anti-AM antibody significantly suppress the growth of established glioblastoma xenografts. The action of AM is specific and is mediated by the calcitonin receptor-like receptor/receptor activity-modifying protein-2 and -3 (CRLR/RAMP2, CRLR/RAMP3). Furthermore, the proangiogenic action of AM on cultured endothelial cells via CRLR/RAMP2 and CRLR/RAMP3 receptors may translate in vivo into enhanced neovascularization and therefore identify AM and its receptors acting as potential new targets for antiangiogenic therapies. PMID- 15888389 TI - [Magnetic resonance spectroscopy in gliomas]. AB - Magnetic resonance spectroscopy (MRS), that may be added to conventional magnetic resonance imaging (MRI) exam exhibit an increasing role in the management of brain tumors. These technique allow quantitative analysis of metabolites, either cell specific, either reflecting physiological and/or pathological process. With a rigorous approach, MRS explore brain metabolism that may improve MRI data in clinical practice in neuro-oncology. Positive diagnosis of brain tumor, differential diagnosis between infiltrative glioma (grade II) and gliomatosis, determination of the limits of tumor infiltration, and distinction between tumor and post-therapeutic images are some of the potential applications of MRS. Ongoing and future studies may also precise the place of MRS in the differential diagnosis between high grade glioma, metastasis and CNS lymphoma, as well as in monitoring therapy in glioma. PMID- 15888390 TI - [Use of the functional imaging modalities in radiation therapy treatment planning in patients with glioblastoma]. AB - Glioblastoma multiforme still remains, at present, the most frequent and deadly primary malignant glioma in adult. Despite safer and larger neurosurgical resections, patients almost always relapse very close or inside the tumor bed. Since more than 20 years, radiation therapy (RT) continue delivering the same dose of 60 Gy in 6 weeks, more precisely guided with CT-scanner and magnetic resonance imaging (MRI) in the treatment position. If morbidity has decreased with "non whole-brain" volumes, RT is nearly always failing locally, as surgery. Until now, all the series evaluating escalating doses (up to 80-90 Gy) in limited volumes have failed. One can really question : is the good dose delivered in the adequate volume? Main goal of new imaging techniques is to better visualize microscopic extension of malignant glioma cells. As based on metabolic principles, areas of abnormalities visualized with functional imaging have a different meaning, often complementary from conventional data. The four evaluated techniques are : magnetic resonance spectroscoy (MRS), functional MRI (fMRI), 18FDG or methionine PET, IMT (123iodine-alpha-methyl-thyrosine) SPECT. Each technique has potential interests and limits, MRS and fMRI appearing the most promising : they have both acceptable spatial resolution and can be executed just after conventional MRI acquisition. Areas of functional abnormalities are only partially including areas of hyperintensity in T1, T2 weighted MRI. It is therefore highly possible that, using it complementary to conventional CT and MRI for RT treatment planning, they add some precious informations; consequently, the very limited efficacy/toxicity ratio could be increased. This hypothesis will only be confirmed by prospective studies registering in parallel both functional and morphological abnormalities, linking them with sites of local recurrence. Once "targeted" the real microscopically invaded areas, one can speculate on new escalating dose studies, delivering RT in "adequate" volumes, combining it with new "targeted" drugs, as already recently demonstrated in head and neck cancers. PMID- 15888391 TI - [New place of the chemotherapy in gliomas]. AB - During these last 25 years, despite numerous phases III studies, standard of treatment in glioblastoma multiforme (GBM) consisted of surgery and post operative radiotherapy, while benefit of chemotherapy was a matter of debate. A phase III study, conducted by EORTC and NCI Canada and involving 573 patients, concluded clearly to the benefit of adding temozolomide during and after radiotherapy as adjuvant treatment. Using this schedule, median survival increase from 12,1 to 14,6 months, and 2 year survival rate from 8 to 26%. In the same time, for anaplastic oligodendroglioma (AO) the phase III study conducted by Cairncross, that compared radiotherapy to a schedule that deliver a chemotherapy with PCV followed by radiotherapy, failed to determine a significant benefit on overall survival, despite the particular chemosensitivity of theses tumors. Moreover, these two studies did underlined impact of biological markers (methylation of the promoter of O6 methylguanine DNA transferase (MGMT) gene for GBM and chromosomes 1p and 19q deletion for AO), that may become decisional markers in the future. We review here the new place of chemotherapy in the adjuvant treatment of GBM and anaplastic gliomas, as well as the impact of these pivotal studies on second lines therapies, and future clinical research. PMID- 15888392 TI - [New therapeutic approaches in glioblastomas]. AB - New therapeutic approaches based on the understanding of brain tumour biology are emerging. Signal transduction inhibitors (mainly targeted against the EGF receptor, the PI3K/Akt or the Ras/Raf pathways), proteases inhibitors and antiangiogenic agents are currently under investigations in clinical trials. The recent development of convection-enhanced delivery technique allows the administration of drugs which do not cross the blood-brain-barrier, such as selective toxins or immunostimulating oligonucleotides. This article reviews these emerging therapies currently under clinical trials in glioblastomas. PMID- 15888393 TI - [Angiogenesis and anti-angiogenic strategies for glioblastoma]. AB - The poor prognosis of patients with glioblastoma multiforme in spite of aggressive conventional anticancer therapies has led to the search for new therapeutic strategies. As glioblastomas are highly vascularized and their growth is angiogenesis-dependent, the inhibition of the sprouting of new capillaries from preexisting blood vessels is one of the most promising therapeutic approaches. Different anti-angiogenic strategies have been developed: inhibition of pro-angiogenic factors and/or receptors and/or downstream cell signaling, inactivation of endothelial cells, inhibition of cellular adhesion molecules and/or extracellular matrix remodeling. Inhibitors of angiogenesis are separated into endogenous inhibitors such as angiostatin, trombospondin or alpha interferon and natural or synthetic inhibitors such as thalidomide, antibodies against angiogenic growth factors or inhibitors of tyrosine kinase receptors. In this review, the majority of experimental studies in glioblastoma models in vivo are summarized and clinical perspectives are discussed. PMID- 15888394 TI - [Participation in the national experiment of cancer diagnosis disclosure in neuro oncology: implications and the perception of the health care team]. AB - Announcement of cancer diagnosis and first contact with medical team in charge of treatment is critical for the patient as well as for the quality of care. The French Cancer Plan include a special focus on these announcement which should be performed in adequate conditions and which should include the presentation of a personalised treatment plan, that reflect multidisciplinary discussion and decision performed before. In order to generalized a formal announcement consultation all over the country in 2005, an experimentation has been performed in 58 centers grouped in 37 projects. Neuro-Oncology Unit of the Assistance Publique-Hopitaux de Marseille did participate to that experimentation. Evaluation of these experiment focus on physician, nurse, and patient perception as well as cost impact and exploration of language during that particular time. Implication of a referent nurse in that time was a strong option of our center. We analyzed the result in 80 patients referred in our center for initial diagnosis (78%) or announcement of recurrence (14%) in a 5 months time. Implication of the nurse appear to be critical for patient confidence and accessibility to information for the all medical team. Organization of a particular time devoted to cancer announcement appear to have a favourable impact on quality of care and patient perception, and constitute a first step to a personalized management of patients with cancer. PMID- 15888395 TI - [Clinical Practice Guidelines 2004. Standards, Options and Recommendations for the management of patient with adenocarcinoma of the stomach: radiotherapy (therapeutic evaluation)]. AB - CONTEXT: The "Standards, Options and Recommendations" (SOR) project, started in 1993, is a collaboration between the Federation of French Cancer Centers (FNCLCC), the 20 French regional cancer centers, and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and the outcome of cancer patients. OBJECTIVES: To elaborate clinical practice guidelines for patients with stomach adenocarcinoma. These recommendations cover the diagnosis, treatment and follow-up of these tumors. METHODS: The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts, with feedback from specialists in cancer care delivery. The Standards, Options and Recommendations are thus based on the best available evidence and expert agreement. RESULTS: This guidelines presents the synthesis of the data concerning the evaluation of the therapeutic ones. The main questions concern the type of gastrectomy to realize (Total Gastrectomy or gastrectomy subtotal), the extent of the lymphadenectomy (D2, D3 versus D1, D3, D2 versus D4) and the role of postoperative chemotherapy and adjuvant concomitant chemoradiotherapy. PMID- 15888396 TI - [Revocation of the BRCA1 patents by the European Office of Patents: a victory over a rights abuse]. PMID- 15888397 TI - Sexual practices, risk perception and knowledge of sexually transmitted disease risk among lesbian and bisexual women. AB - CONTEXT: Sexually transmitted diseases (STDs) can be spread between female sex partners, probably through the exchange of cervicovaginal fluid and direct mucosal contact. Additionally, lesbians have a high prevalence of bacterial vaginosis, which may represent an STD in this population. However, few data on sexual practices or perceived STD risk among lesbians are available to guide development of interventions aimed at reducing the risk. METHODS: To inform the development of a safer-sex intervention for women who have sex with women, focus group discussions were conducted with 23 lesbian and bisexual women aged 18-29. Topics included sexual practices, STD transmission and prevention, and knowledge about bacterial vaginosis. RESULTS: Although six participants had had bacterial vaginosis and three an STD, women reported little use of preventive measures with female partners (washing hands, using rubber gloves and cleaning sex toys). Participants said that vaginal penetrative practices using sex toys and fingers or hands are common, and that partners frequently share sex toys during a sexual encounter, generally without condoms. Knowledge of potential for STD transmission between women, and of bacterial vaginosis, was limited. Participants viewed use of barrier methods (gloves or condoms) as acceptable, provided that there is a reason (usually STD-focused) to use them and that they are promoted in the context of sexual health and pleasure. CONCLUSIONS: Safer-sex messages aimed at lesbian and bisexual women should emphasize the plausibility of STD transmission between women, personal responsibility and care for partners' well-being; should target common sexual practices; and should promote healthy sexuality. PMID- 15888398 TI - Sexual intentions of black preadolescents: associations with risk and adaptive behaviors. AB - CONTEXT: Adolescent sexual activity in the United states is prevalent and occurring increasingly early, particularly among minority groups. Other risk behaviors (e.g., alcohol consumption) often co-occur with sexual behavior. By examining the association of risk and adaptive behaviors with precursors of sexual behavior--specifically, sexual intentions--it may be possible to identify preadolescents who are at increased risk for early sexual initiation. METHODS: Data from 1,090 black fourth and fifth graders and their parents from the Parents Matter! Program were used in logistic regression analyses to assess covariation between preadolescents' risk and adaptive behaviors, and their intentions to initiate sexual intercourse in the next year. RESULTS: Risk and adaptive behaviors, as reported by both preadolescents and parents, were associated with sexual intentions; the findings were not qualified by youth's gender. Alcohol consumption and having been in trouble with the police were the primary youth reported risk behaviors associated with the odds of intending to have intercourse (odds ratios, 2.3 and 1.8); the preadolescent's being in trouble at home was the primary parent-reported risk behavior (2.1). In both sets of reports, performing well on schoolwork was associated with reduced odds of intending to engage in sex (0.5 - 0.6). CONCLUSIONS: Risk and adaptive behaviors are markers of sexual intentions among black preadolescents. Prevention programs can use these behaviors to identify black youth who may be at high risk for early sexual initiation. PMID- 15888399 TI - Emergency contraceptive pills: dispensing practices, knowledge and attitudes of South Dakota pharmacists. AB - CONTEXT: Despite a decision by the Food and Drug Administration to deny over-the counter status to emergency contraceptive pills, pharmacists play a crucial role in a woman's access to this medication, especially in areas with large rural populations. Pharmacists' knowledge about and attitudes toward emergency contraceptive pills may affect whether pharmacies carry the medication and whether individual pharmacists dispense it. METHODS: In October 2003, all registered pharmacists living and working in South Dakota were mailed a survey to assess their dispensing practices for, knowledge about, and attitudes toward emergency contraceptive pills. Data for 501 respondents were analyzed through chi square testing and multivariate logistic regression. RESULTS: Fifty-four percent of respondents worked in pharmacies that carried emergency contraceptive pills. Of these, 67% had dispensed the medication in 2003, and 24% were not comfortable providing customer counseling about the method. Thirty-seven percent of all pharmacists did not understand its mechanism of action; 43% and 21%, respectively, incorrectly answered questions about the medication's link to birth defects and health risks. Only 5% correctly answered all three questions. Eighty four percent of surveyed pharmacists believed that the medication should not be made available over the counter. Multivariate analysis showed that knowledge of emergency contraception and support for over-the-counter status were relatively low among pharmacists working in small communities. CONCLUSIONS: The education of pharmacists about emergency contraceptive pills must be strengthened to ensure that women receive accurate medical information and access to all contraceptive services. PMID- 15888400 TI - Youth assets and sexual risk behavior: the importance of assets for youth residing in one-parent households. AB - CONTEXT: Youth assets are associated with a reduction in sexual risk behavior; however, little is known about this association among youth at high risk of engaging in unsafe behavior, such as those in one-parent households. METHODS: In home interviews were used to collect data from 1,253 inner-city teenagers and their parents. Multivariate logistic regressions were conducted separately for youth from one- and two-parent households to assess relationships between youth assets and four behaviors related to sexual risk: never having had sexual intercourse, not being currently sexually active, having delayed intercourse until age 17, and having used birth control at last intercourse. RESULTS: Among youth living in one-parent households, those with the aspirations for the future, good health practices (exercise/nutrition), peer role models and family communication assets had significantly elevated odds of reporting one of the behaviors examined (odds ratios, 1.8-7.3). The peer role models asset also interacted with parental education to significantly predict an absence of current sexual activity for youth in one-parent households (21.2). Among youth living in two-parent households, community involvement was linked to increased odds of never having had sex (1.9), but no other significant relationships were found. Youths' total number of assets significantly predicted three of the four behaviors among youth in one-parent households (1.2-1.8), but predicted only sexual inexperience among those in two-parent households (1.4). CONCLUSION: Future research should more fully investigate the role of family structure in relationships between youth assets and risk behaviors. Certain youth assets may be particularly effective in reducing sexual risk behavior among youth in one parent households. PMID- 15888401 TI - Correlates of partner-specific condom use intentions among incarcerated women in Rhode Island. AB - CONTEXT: Few studies of incarcerated women have examined potential associations between risky sexual behavior and relationship context factors; thus, little is known about the correlates of intentions to use condoms with main and casual partners in this underserved population. METHODS: A sample of 221 women incarcerated in a Rhode Island Department of Corrections facility in 2002-2003 were interviewed. Multiple linear regression analysis was performed to assess associations between selected demographic, psychosocial and behavioral variables and participants' reported intentions to use condoms with main and casual sexual partners in the first six months after their release. RESULTS: Condom use at last sex with a main partner, sexually transmitted disease (STD) history, no strong desire to currently be pregnant, belief that others influence one's health and perceived STD risk were positively associated with women's intention to use condoms with main partners. Pregnancy history was negatively associated with intention to use condoms with a main partner. Condom use at last sex with a casual partner was positively associated with intention to use condoms with casual partners, whereas binge drinking and believing in the role of chance in determining one's health were negatively associated with intention to use condoms with casual partners. CONCLUSIONS: Whether incarcerated women define a partner as main or casual may influence their decisions about the need to protect themselves by using condoms. Programs that focus on the importance of condom use with all partners could greatly benefit incarcerated women and the communities to which they return. PMID- 15888402 TI - Opportunities for action: addressing Latina sexual and reproductive health. PMID- 15888403 TI - Cultural sensitivity and research involving sexual minorities. PMID- 15888404 TI - Confronting the 'sugar daddy' stereotype: age and economic asymmetries and risky sexual behavior in urban Kenya. AB - CONTEXT: "Sugar daddy" relationships, which are characterized by large age and economic asymmetries between partners, are believed to be a major factor in the spread of HIV in Sub-Saharan Africa. Information is needed about sugar daddy partnerships-and about age and economic asymmetries more generally-to determine how common they are and whether they are related to unsafe sexual behavior. METHODS: The sample comprised 1,052 men aged 21-45 who were surveyed in Kisumu, Kenya, in 2001. Data on these men and their 1,614 recent non-marital partnerships were analyzed to calculate the prevalence of sugar daddies and sugar daddy relationships, as well as a range of age and economic disparities within non marital partnerships. Logistic regression models were constructed to assess relationships between condom use at last sexual intercourse and various measures of age and economic asymmetry. RESULTS: The mean age difference between non marital sexual partners was 5.5 years, and 47% of men's female partners were adolescents. Fourteen percent of partnerships involved an age difference of at least 10 years, and 23% involved more than the mean amount of male-to-female material assistance. Men who reported at least one partnership with both these characteristics were defined as sugar daddies and made up 5% of the sample; sugar daddy relationships accounted for 4% of partnerships. Sugar daddy partnerships and the largest age and economic asymmetries we constructed were associated with decreased odds of condom use. CONCLUSIONS: Although sugar daddy relationships are not as pervasive as generally assumed, age and economic asymmetries in non marital partnerships are relatively common. All these types of asymmetries are associated with nonuse of condoms. Increasing women's power within asymmetric sexual relationships could improve their ability to negotiate safer sexual behaviors, such as condom use. PMID- 15888405 TI - The impact of menstrual side effects on contraceptive discontinuation: findings from a longitudinal study in Cairo, Egypt. AB - CONTEXT: Although many research studies have documented the relationship between menstrual side effects of contraceptives and discontinuation of use, few have sought to identify factors that predispose women to discontinue because of changes in bleeding patterns. Such information is important to enable family planning providers to better help women and couples choose appropriate methods and use them successfully. METHODS: Forty-eight women participating in six focus group discussions described their experiences using the IUD, the hormonal implant or the three-month injectable. Subsequently, 259 women using one of these methods for the first time were followed for up to 18 months to determine patterns of menstrual bleeding and perceptions of menstrual cycle change over time. Multivariable analytical methods were used to examine the associations between selected measures and method discontinuation. RESULTS: Contraceptive discontinuation differed by method: Nearly 70% of injectable users had stopped using their chosen method after one year, compared with 34% of IUD users and 10% of implant users. Before initiating a method, women reported an average of five bleeding days per cycle. During the first six months of use, IUD users reported an average of six days of bleeding per cycle; injectable and implant users reported 11-12. In multivariable models, each additional day of bleeding was significantly associated with a 2-4% increase in discontinuation, depending on method type. Among IUD users, women whose husbands knew that they had visited a clinic to initiate a method were less likely than others to discontinue method use (hazard ratio, -1.9). Age was significantly associated with decreased discontinuation among implant users. CONCLUSION: Counseling about bleeding and other side effects should be tailored to women's personal contexts and contraceptive experiences. PMID- 15888406 TI - Risk perception and condom use among married or cohabiting couples in KwaZulu Natal, South Africa. AB - CONTEXT: Most HIV prevention efforts focus on premarital and extramarital sexual behavior, but in areas with high HIV prevalence the protective needs of married and cohabiting couples are just as great and often go unmet. Condom use by these couples is generally low, with resistance from men and cultural norms commonly cited as barriers to increased use. METHODS: A household survey was conducted in an urban and a rural area in KwaZulu-Natal, South Africa, in 1999-2000. From this survey, matched partners in 238 marital or cohabiting relationships were independently interviewed about condom use and attitudes toward condoms, knowledge of AIDS/HIV risk and self-efficacy in preventing HIV infection. Logistic regression analysis was used to assess relationships between condom use and selected demographic and HIV prevention characteristics. RESULTS: Although couples' knowledge of condoms and where to obtain them was very high, only 15% of men and 18% of women reported consistent or occasional use. The level of use was 8% and 11% among men and women, respectively, in rural, less educated couples, and 29% and 34% among men and women in urban, more educated couples. A majority of urban women had favorable attitudes toward condoms, and they also reported higher self-efficacy regarding HIV prevention than did rural women. A woman's perceived risk of HIV infection from her partner was the most powerful predictor of condom use (odds ratio, 4.0). CONCLUSIONS: The common belief that men's resistance to condom use within stable relationships cannot be overcome may be exaggerated. HIV prevention programs should address the reproductive health needs of these couples. PMID- 15888407 TI - Factors associated with use of the female condom in Zimbabwe. AB - CONTEXT: Because women can initiate use of the female condom, the method is believed to make it easier for women to protect themselves against sexually transmitted infections (STIs), including HIV infection. Evidence is lacking about factors associated with trying the female condom and using it consistently. METHODS: A sample of 1,740 sexually active consumers visiting retail outlets in urban Zimbabwe that sell male or female condoms were surveyed in 1998, one year after a social marketing campaign had begun. Logistic regression analyses were conducted to assess factors associated with ever-use of the female condom and consistent use (always or often) with marital and regular non-marital partners. RESULTS: Perceived ease of use and affordability of the product and prior use of the male condom were associated with men's and women's ever-use. Consistent use with marital partners was negatively associated with reporting multiple partners in the past year (odds ratio, 0.3) and positively associated with using the device for pregnancy prevention (5.4) and previously using the male condom (8.0). Consistent use with regular non-marital partners was associated with numerous variables, including perceived ease of use (1.9) and effectiveness for STI prevention (3.8), low HIV risk perception (2.4), and use for pregnancy (2.9) and STI (2.3) prevention. CONCLUSIONS: Perceived affordability and ease of use may encourage couples to try the female condom but may not lead to consistent use. Because the reasons for use can vary between marital and non-marital relationships, the female condom may need to be positioned differently for different target populations. PMID- 15888408 TI - Does 'CNN' (condoms, needles and negotiation) work better than 'ABC' (abstinence, being faithful and condom use) in attacking the AIDS epidemic? PMID- 15888409 TI - Molecular preservation in Late Cretaceous sauropod dinosaur eggshells. AB - Exceptionally preserved sauropod eggshells discovered in Upper Cretaceous (Campanian) deposits in Patagonia, Argentina, contain skeletal remains and soft tissues of embryonic Titanosaurid dinosaurs. To preserve these labile embryonic remains, the rate of mineral precipitation must have superseded post-mortem degradative processes, resulting in virtually instantaneous mineralization of soft tissues. If so, mineralization may also have been rapid enough to retain fragments of original biomolecules in these specimens. To investigate preservation of biomolecular compounds in these well-preserved sauropod dinosaur eggshells, we applied multiple analytical techniques. Results demonstrate organic compounds and antigenic structures similar to those found in extant eggshells. PMID- 15888410 TI - Tracking butterfly flight paths across the landscape with harmonic radar. AB - For the first time, the flight paths of five butterfly species were successfully tracked using harmonic radar within an agricultural landscape. Until now, butterfly mobility has been predominantly studied using visual observations and mark-recapture experiments. Attachment of a light-weight radar transponder to the butterfly's thorax did not significantly affect behaviour or mobility. Tracks were analysed for straightness, duration, displacement, ground speed, foraging and the influence of linear landscape features on flight direction. Two main styles of track were identified: (A) fast linear flight and (B) slower nonlinear flights involving a period of foraging and/or looped sections of flight. These loops potentially perform an orientation function, and were often associated with areas of forage. In the absence of forage, linear features did not provide a guiding effect on flight direction, and only dense treelines were perceived as barriers. The results provide tentative support for non-random dispersal and a perceptual range of 100-200 m for these species. This study has demonstrated a methodology of significant value for future investigation of butterfly mobility and dispersal. PMID- 15888411 TI - Cone topography and spectral sensitivity in two potentially trichromatic marsupials, the quokka (Setonix brachyurus) and quenda (Isoodon obesulus). AB - The potential for trichromacy in mammals, thought to be unique to primates, was recently discovered in two Australian marsupials. Whether the presence of three cone types, sensitive to short- (SWS), medium- (MWS) and long- (LWS) wavelengths, occurs across all marsupials remains unknown. Here, we have investigated the presence, distribution and spectral sensitivity of cone types in two further species, the quokka (Setonix brachyurus) and quenda (Isoodon obesulus). Immunohistochemistry revealed that SWS cones in the quokka are concentrated in dorso-temporal retina, while in the quenda, two peaks were identified in naso ventral and dorso-temporal retina. In both species, MWS/LWS cone spatial distributions matched those of retinal ganglion cells. Microspectrophotometry (MSP) confirmed that MWS and LWS cones are spectrally distinct, with mean wavelengths of maximum absorbance at 502 and 538 nm in the quokka, and at 509 and 551 nm, in the quenda. Although small SWS cone outer segments precluded MSP measurements, molecular analysis identified substitutions at key sites, accounting for a spectral shift from ultraviolet in the quenda to violet in the quokka. The presence of three cone types, along with previous findings in the fat tailed dunnart and honey possum, suggests that three spectrally distinct cone types are a feature spanning the marsupials. PMID- 15888412 TI - Biosonar behaviour of free-ranging porpoises. AB - Detecting objects in their paths is a fundamental perceptional function of moving organisms. Potential risks and rewards, such as prey, predators, conspecifics or non-biological obstacles, must be detected so that an animal can modify its behaviour accordingly. However, to date few studies have considered how animals in the wild focus their attention. Dolphins and porpoises are known to actively use sonar or echolocation. A newly developed miniature data logger attached to a porpoise allows for individual recording of acoustical search efforts and inspection distance based on echolocation. In this study, we analysed the biosonar behaviour of eight free-ranging finless porpoises (Neophocaena phocaenoides) and demonstrated that these animals inspect the area ahead of them before swimming silently into it. The porpoises inspected distances up to 77 m, whereas their swimming distance without using sonar was less than 20 m. The inspection distance was long enough to ensure a wide safety margin before facing real risks or rewards. Once a potential prey item was detected, porpoises adjusted their inspection distance from the remote target throughout their approach. PMID- 15888413 TI - Avian uncoupling protein expressed in yeast mitochondria prevents endogenous free radical damage. AB - The longevity of birds is surprising since they exhibit high metabolic rates and elevated blood sugar levels compared with mammals of the same body size, which presumably expose them to higher rates of free oxygen radical production, which is implicated in accelerated senescence. Uncoupling proteins (UCPs) are transporters of the inner mitochondrial membrane and their physiological activity is still a subject of debate. Avian UCP was found in birds but data on its activity are scarce. Avian UCP (Gallus gallus) was overexpressed in yeast and we assessed its ability to prevent mitochondrial reactive oxygen species (ROS) production by measuring ROS damage (aconitase activity) and antioxidant defences (MnSOD activity). We show that avian UCP protects yeast mitochondria against the deleterious impact of ROS, but without stimulation of superoxide dismutase activity. Avian UCP protein was specifically immunodetected and retinoic acid, which belongs to the carotenoid family, was found to trigger its activity. These data show that avian UCP basal activity protects against ROS damage. However, when activated by retinoic acid, avian UCP can also operate as the mammalian thermogenic UCP1. The hypothesis that avian UCP activities are state- and species dependent is further discussed. PMID- 15888414 TI - Adaptive significance of avian beak morphology for ectoparasite control. AB - The beaks of Darwin's finches and other birds are among the best known examples of adaptive evolution. Beak morphology is usually interpreted in relation to its critical role in feeding. However, the beak also plays an important role in preening, which is the first line of defence against harmful ectoparasites such as feather lice, fleas, bugs, flies, ticks and feather mites. Here, we show a feature of the beak specifically adapted for ectoparasite control. Experimental trimming of the tiny (1-2 mm) maxillary overhang of rock pigeons (Columba livia) had no effect on feeding efficiency, yet triggered a dramatic increase in feather lice and the feather damage they cause. The overhang functions by generating a shearing force against the tip of the lower mandible, which moves forward remarkably quickly during preening, at up to 31 timesper second. This force damages parasite exoskeletons, significantly enhancing the efficiency of preening for parasite control. Overhangs longer than the natural mean of 1.6mm break significantly more often than short overhangs. Hence, stabilizing selection will favour overhangs of intermediate length. The adaptive radiation of beak morphology should be re-assessed with both feeding and preening in mind. PMID- 15888415 TI - Wave energy and swimming performance shape coral reef fish assemblages. AB - Physical factors often have an overriding influence on the distribution patterns of organisms, and can ultimately shape the long-term structure of communities. Although distribution patterns in sessile marine organisms have frequently been attributed to functional characteristics interacting with wave-induced water motion, similar evidence for mobile organisms is lacking. Links between fin morphology and swimming performance were examined in three diverse coral reef fish families from two major evolutionary lineages. Among-habitat variation in morphology and performance was directly compared with quantitative values of wave induced water motion from seven coral reef habitats of different depth and wave exposure on the Great Barrier Reef. Fin morphology was strongly correlated with both field and experimental swimming speeds in all three families. The range of observed swimming speeds coincided closely with the magnitude of water velocities commonly found on coral reefs. Distribution patterns in all three families displayed highly congruent relationships between fin morphology and wave-induced water motion. Our findings indicate a general functional relationship between fin morphology and swimming performance in labriform-swimming fishes, and provide quantitative evidence that wave energy may directly influence the assemblage structure of coral reef fishes through interactions with morphology and swimming performance. PMID- 15888416 TI - A comparative analysis of the evolutionary relationship between diet and enzyme targeting in bats, marsupials and other mammals. AB - The subcellular distribution of the enzyme alanine:glyoxylate aminotransferase (AGT) in the livers of different mammals appears to be related to their natural diets. Thus, AGT tends to be mitochondrial in carnivores, peroxisomal in herbivores, and both mitochondrial and peroxisomal in omnivores. To what extent this relationship is an incidental consequence of phylogenetic structure or an evolutionarily meaningful adaptive response to changes in dietary selection pressure is unknown. In order to distinguish between these two possibilities, we have determined the subcellular distribution of AGT in the livers of 22 new mammalian species, including members of three orders not studied before. In addition, we have analysed the statistical relationship between AGT distribution and diet in all 77 mammalian species, from 12 different orders, for which the distribution is currently known. Our analysis shows that there is a highly significant correlation between AGT distribution and diet, independent of phylogeny. This finding is compatible with the suggestion that the variable intracellular targeting of AGT is an adaptive response to episodic changes in dietary selection pressure. To our knowledge, this is the first example of such a response being manifested at the molecular and cellular levels across the breadth of Mammalia. PMID- 15888417 TI - Flexibility in assessment of prey cues: frog-eating bats and frog calls. AB - Predators use cues associated with their prey to assess prey quality and to avoid consuming poisonous prey. Considerable attention has been given to predators' use of aposematic cues to assess prey quality, but little is known about predators that eavesdrop on prey cues that are not intended for them. Here we investigate the prey-cue/prey-quality associations of a predator that eavesdrops on the sexual advertisement signals of its prey. Stability is expected in prey-cue/prey quality associations when mistakes in prey assessment are lethal. Conversely, flexibility is possible when mistakes are less costly. Predators that must respond to temporal and spatial fluctuations in prey availability should be more flexible in their assessment of prey quality. Given these predictions, we examined flexibility in the ability of wild-caught bats to reverse prey-cue/prey quality associations for a preferred prey and a poisonous one. We found that the predatory bat, Trachops cirrhosus, has a heretofore undescribed ability to reverse its evaluations of the cues that signal preferred prey. PMID- 15888418 TI - African endemics span the tree of songbirds (Passeri): molecular systematics of several evolutionary 'enigmas'. AB - The deep divergence between the African endemic passerines Picathartidae (rockfowl Picathartes and rockjumpers Chaetops, four species) and the Passerida (ca. 3500 species) suggests an older history of oscines on the African continent than has previously been assumed. In order to determine whether any additional, unexpectedly deep lineages occur in African endemic songbirds, 29 species- including 10 enigmatic focal taxa endemic to southern Africa--were added to a large nuclear sequence dataset gathered from oscine songbirds (Passeri). Phylogenetic analyses of these data resolve many long-standing questions about the affinities of these birds, not all of which were predicted by traditional approaches. The application of a molecular clock indicates that most basal divergences in Passerida occurred in the middle to late Eocene, with divergences between African and Australasian core corvoids occurring somewhat later in the early Miocene. Consistent with inferences for mammals, divergences between Malagasy endemic passerines and their mainland relatives suggests an asynchronous colonization history. This emerging phylogenetic picture reveals that relationships within Old World families are highly informative regarding the early dispersal and radiation of songbirds out of Gondwana. Future analyses will depend on improving resolution of higher-level phylogenetic relationships among these groups, and increasing the density of taxon sampling within them. PMID- 15888419 TI - Maternal rank and local resource competition do not predict birth sex ratios in wild baboons. AB - We test two models of adaptive adjustment of birth sex ratios that are expected to apply to Cercopithecine primate species. It has been predicted that when maternal investment differentially influences the reproductive success of male and female offspring, females in good condition will bias investment in favour of the sex that gains the greatest fitness returns from additional investment. This hypothesis was subsequently amended to take into account the effects of local resource competition on maternal investment strategies of primate females. This body of theory has been applied to primates with contradictory results, prompting some to question the conclusion that primate females facultatively adjust birth sex ratios in an adaptive manner. Here, we present a meta-analysis of the relationship between maternal rank, birth sex ratios and local resource competition in 36 groups of wild savannah baboons, Papio cynocephalus. The results do not support predictions derived from either model of facultative sex ratio adjustment, and we conclude that there is currently no evidence that baboon birth sex ratios are adjusted in an adaptive manner. PMID- 15888420 TI - The geological history of deep-sea colonization by echinoids: roles of surface productivity and deep-water ventilation. AB - The origins and geological history of the modern fauna of deep-sea echinoids is explored using a combination of palaeontological and molecular data. We demonstrate that, whereas generalist omnivores have migrated into the deep sea in low numbers over the past 200 Myr, there was a short time-interval between approximately 75 and 55 Myr when the majority of specialist detritivore clades independently migrated off-shelf. This coincides with a marked increase in seasonality, continental run-off and surface water productivity, and suggests that increasing organic carbon delivery into ocean basins was an important controlling factor. Oceanic anoxic events, by contrast, appear to have played a subsidiary role in controlling deep-sea diversity. PMID- 15888421 TI - Inferences about information flow and dispersal for spatially extended population systems using time-series data. AB - This work explores an information-theoretic approach to drawing inferences about coupling of spatially extended ecological populations based solely on time-series of abundances. The efficacy of the approach, time-delayed mutual information, was explored using a spatially extended predator-prey model system in which populations at different patches were coupled via diffusive movement. The approach identified the relative magnitude and direction of information flow resulting from animal movement between populations, the change in information flow as a function of distance separating populations, and the diffusive nature of the information flow. In addition, when the diffusive movement was eliminated from the model, mutual information correctly provided no evidence of information flow, even when population synchrony was generated by a common environmental driving function. Thus, for this model system, time-delayed mutual information was useful in discriminating between the Moran effect and animal movement as causes of population synchrony, as well as in characterizing dispersal in terms of direction, relative speed and diffusive nature. PMID- 15888422 TI - A summary of the FDA-NIMH-MATRICS workshop on clinical trial design for neurocognitive drugs for schizophrenia. AB - OBJECTIVE: On April 23, 2004, a joint meeting of the FDA, NIMH, MATRICS investigators, and experts from academia and the pharmaceutical industry was convened to develop guidelines for the design of clinical trials of cognitive enhancing drugs for neurocognitive impairments in patients with schizophrenia. METHOD: Experts were asked to address specific questions relating to clinical trial design of adjunctive/co-treatment and broad spectrum agents. At the workshop, experts reviewed relevant evidence before offering the discussion panel proposed guidelines for a given subset of questions. The discussion panel, which consisted of presenters and representatives from FDA, NIMH, academia, and industry, deliberated to reach consensus on suggested guidelines. When evidence was insufficient, suggested guidelines represent the opinion of a cross-section of the presenters and discussion panel. RESULTS: Guidelines were developed for inclusion criteria, the use of co-primary outcome measures, and statistical approaches for study design. Consensus was achieved regarding diagnostic and concomitant medication inclusion criteria and on the use of cognitive screening measures. A key guideline was to limit the trial to patients in the residual phase of their illness, who have a predefined level of positive, negative, and affective symptoms. The most difficult issues were the feasibility of including a co-primary measure of functional improvement and the choice of comparator agent for a trial of a broad spectrum agent (with antipsychotic and cognitive-enhancing effects). CONCLUSIONS: The suggested guidelines represent reasonable starting points for trial design of cognitive-enhancing drugs, with the understanding that new data, subsequent findings, or other methodological considerations may lead to future modifications. PMID- 15888423 TI - "Theory of mind" in schizophrenia: a review of the literature. AB - The term theory of mind (ToM) refers to the capacity to infer one's own and other persons' mental states. A substantial body of research has highlighted the evolution of ToM in nonhuman primates, its emergence during human ontogeny, and impaired ToM in a variety of neuropsychiatric disorders, including schizophrenia. There is good empirical evidence that ToM is specifically impaired in schizophrenia and that many psychotic symptoms-for instance, delusions of alien control and persecution, the presence of thought and language disorganization, and other behavioral symptoms-may best be understood in light of a disturbed capacity in patients to relate their own intentions to executing behavior, and to monitor others' intentions. However, it is still under debate how an impaired ToM in schizophrenia is associated with other aspects of cognition, how the impairment fluctuates with acuity or chronicity of the schizophrenic disorder, and how this affects the patients' use of language and social behavior. In addition to these potential research areas, future studies may also address whether patients could benefit from cognitive training in this domain. PMID- 15888424 TI - The schizophrenic experience: taken out of context? AB - A currently favored cognitive model of the abnormal behaviors and experiences characteristic of schizophrenia suggests that they may be linked to a disturbance in the effects of context. The present article reviews some of the relevant literature, noting the wide range of experimental paradigms that have been employed. This range of paradigms is both a strength and a potential weakness of the literature because it raises complex issues of definition and the need to distinguish the various ways that context may influence behaviors. This influence may be crucially dependent on specific task parameters. Despite this, a number of phenomena can be plausibly related to changes in the way that context operates: delusions, disorganization, hallucinations, and the loss of a sense of personal identity. Potential links with the neural bases of the disorder are indicated. PMID- 15888425 TI - Cognitive strategies versus self-management skills as adjunct to vocational rehabilitation. AB - Cognitive dysfunctions and negative symptoms are "rate-limiting factors" for community outcome and response to psychosocial intervention in people with schizophrenia. Therefore, two cognitive-behavioral group therapies were developed computer-assisted cognitive strategy training (CAST) and training of self management skills for negative symptoms (TSSN)-to target these barriers to rehabilitation readiness. One hundred thirty-eight DSM-IV schizophrenia inpatients on a rehabilitation ward were randomly assigned to CAST plus vocational rehabilitation, TSSN plus vocational rehabilitation, or vocational rehabilitation alone. CAST included computer-based training in coping strategies focusing on deficits in attention, verbal memory, and planning. TSSN focused on social withdrawal/social anhedonia, lack of drive/volition, and affect flattening using techniques such as time scheduling, mastery, and pleasure techniques. Treatment outcome was assessed at intake and at discharge after 8 weeks. Analyses of covariance controlling for basis-level functioning demonstrated that patients receiving CAST plus vocational rehabilitation showed greater improvement on attention and verbal memory but not on planning ability. Patients receiving TSSN plus vocational rehabilitation failed to demonstrate improvement in negative symptoms. CAST plus vocational rehabilitation was found to be associated with a higher rate of successful job placement at the 12-month followup interval. Hierarchical logistic regression analyses demonstrated that improvement in short- and long-term verbal memory predicted a higher proportion of variance of successful job placement in the followup than pretreatment history of employment alone. Cognitive training as an adjunct to inpatient vocational rehabilitation demonstrated cognitive improvement, which was found to be associated with successful job placement in the followup. TSSN's efficacy was less clear; reasons for this uncertainty are provided. PMID- 15888426 TI - Learning potential and the prediction of work skill acquisition in schizophrenia. AB - This study examined whether a measure of learning potential could predict work skill acquisition in schizophrenia beyond the prediction offered by a single administration assessment. Fifty-seven outpatients with schizophrenia or schizoaffective disorder completed a test-train-test version of the Wisconsin Card Sorting Test as a measure of their learning potential. The outpatients were randomly assigned to training by errorless learning or conventional instruction on two work skills (index card filing and toilet tank assembly). Work skills were assessed both immediately and 3 months after training. Generally, patients with high learning potential performed better on the work skill tasks. Learning potential explained an additional 15 percent of variance beyond single administration assessment in participants' accuracy immediately after work skill training and an additional 13 percent of variance in participants' accuracy 3 months after training. These findings indicate that measures of learning potential contribute to the prediction of work skill acquisition, going beyond the predictive power of single administration assessments. PMID- 15888427 TI - The micro-module learning tests: work-sample assessments of responsiveness to skills training. AB - While outcome of psychiatric rehabilitation has been successfully predicted by cognitive tests, efforts to design a measure to assess responsiveness to rehabilitation have been lacking. In this report, we describe the rationale for and development of a face-valid measure of responsivity to the three core components of skills training: responsiveness to verbal instruction, ability to learn from viewing the behavior of a model, and ability to demonstrate skills observed during a subsequent role-play. Seven alternate forms of the new measure, called the Micro-Module Learning Test (MMLT), demonstrated adequate internal consistency and alternate-form reliability. We also present results from four studies in which the MMLT was used to collect normative data as well as data on relationships with symptoms, cognitive tests, and treatment outcome. Results indicate that the MMLT is associated with cognitive factors found to predict treatment outcome in prior studies (e.g., verbal memory and fluency), as well as lesser investigated functions such as theory of mind ability. In addition, MMLT scores were correlated negatively with psychotic disorganization and positively with performance during a full-length skills training group. The MMLT appears to be a reliable and valid measure for rapidly assessing responsiveness to skills training procedures. PMID- 15888428 TI - Indicators of genetic liability to schizophrenia: a sibling study of neuropsychological performance. AB - Despite clear evidence of important genetic influences on schizophrenia, identifying the genes involved has been difficult because of the genetic complexity of the phenotype. The use of additional phenotypic measures that are more sensitive to the genetic liability than is the clinical diagnosis should enhance the power to detect small individual genetic effects. The present study assessed the neuropsychological performance of 30 male schizophrenia probands, 30 of their unaffected male siblings, and 20 well controls matched on age, sex, and education in order to identify measures that may be particularly sensitive to the genetic liability to schizophrenia and thus may be useful in gene mapping studies. Siblings showed impaired neuropsychological performance compared to controls on four out of the five measures used. Additional results suggested that Trails B was especially effective at discriminating index siblings from controls, thus supporting its potential utility as a candidate quantitative phenotype to aid in gene mapping studies of the disorder. PMID- 15888429 TI - Enhanced vividness of mental imagery as a trait marker of schizophrenia? AB - We assessed the vividness of mental imagery in schizophrenia patients in the context of psychopathology and cognitive abilities. A questionnaire on the vividness of mental imagery (Questionnaire Upon Mental Imagery [QMI]) and a hallucination scale were administered to 50 patients with paranoid schizophrenia. The related perceptual and cognitive skills, general intelligence level, and psychomotor speed were measured as covariates with a battery of performance tests. All measures were statistically compared to a group of 50 age- and sex matched healthy controls. The schizophrenia group obtained higher values both for vividness of imagery and occurrence of hallucinations. These differences were independent of general intelligence and psychomotor speed and did not correlate with individual psychopathology. The correlation between the hallucination and imagery scales themselves was very low. These results suggest that patients with schizophrenia experience a significantly greater vividness of mental imagery than healthy controls, which does not seem to be an effect of other group differences or individual psychopathology (e.g., frequency of hallucinations). Vividness of mental imagery might thus prove to be an independent trait marker of schizophrenia. PMID- 15888430 TI - Schizotypy and conditional reasoning. AB - This study investigated the role of reasoning biases in delusion formation and maintenance. Reasoning judgments have been shown to be influenced by prior knowledge, beliefs, and experience--that is, information stored in semantic memory. It was hypothesized that high schizotypes would exhibit a "jump to conclusions" style of reasoning as a result of not using implicit information concerned with cause-effect relationships. Research into human reasoning has traditionally adopted logic as a normative framework to assess human reasoning. Conditional inference tasks are direct tests of logical performance, and we employed an established design that depends upon the reasoner's ability to access and use implicit information. In an effort to negate some of the difficulties of research with schizophrenia patients, schizotypy measures were employed in a normal population. The results confirmed that high scorers on one dimension of schizotypy (Impulsive Nonconformity) failed to take account of the number of counterexamples that characterized the cause-effect conditional statement. These observations supported previous research demonstrating a jump to conclusions style of reasoning that it has been suggested plays a role in the formation and maintenance of delusions. Furthermore, these findings suggest a possible link between semantic memory and reasoning biases. PMID- 15888431 TI - Cortical cholinergic transmission and cortical information processing in schizophrenia. AB - Models of the neuronal mediation of psychotic symptoms traditionally have focused on aberrations in the regulation of mesolimbic dopaminergic neurons, via their telencephalic afferent connections, and on the impact of abnormal mesolimbic activity for functions of the ventral striatum and its pallidal-thalamic-cortical efferent circuitry. Repeated psychostimulant exposure models major aspects of the sensitized activity of ventral striatal dopaminergic transmission that is observed in patients exhibiting psychotic symptoms. Based on neuroanatomical, neurochemical, and behavioral data, the hypothesis that an abnormally reactive cortical cholinergic input system represents a necessary correlate of a sensitized mesolimbic dopaminergic system is discussed. Moreover, the abnormal cognitive mechanisms that contribute to the development of psychotic symptoms are attributed specifically to the aberrations in cortical cholinergic transmission and to its consequences on the top-down regulation of sensory and sensory associational input functions. Experimental evidence from studies demonstrating repeated amphetamine-induced sensitization of cortical cholinergic transmission and the ability of antipsychotic drugs to normalize the activity of cortical cholinergic inputs, and from experiments indicating the attentional consequences of manipulations that increase the excitability of cortical cholinergic inputs, supports this hypothesis. Relevant human neuropathological and psychopharmacological data are discussed, and the implications of an abnormally regulated cortical cholinergic input system for pharmacological treatment strategies are addressed. Given the role of cortical cholinergic inputs in gating cortical information processing, even subtle changes in the regulation of this cortexwide input system that represent a necessary transsynaptic consequence of sensitized mesolimbic dopaminergic transmission profoundly contribute to the neuronal mediation of psychotic symptoms. PMID- 15888432 TI - Construct validity of the animal latent inhibition model of selective attention deficits in schizophrenia. AB - Latent inhibition (LI) is demonstrated when a previously unattended/inconsequential stimulus is less effective in a new learning situation than a novel stimulus. In rats and humans, LI is reduced by dopamine agonists and increased by dopamine antagonists. In addition, LI is attenuated in actively psychotic schizophrenia patients, thus conferring strong predictive validity to the animal LI preparation for schizophrenia. However, the validity of the attentional construct in the LI model of schizophrenia dysfunction depends on confirming two assumptions: that animal and human LI share a common process, and that the process is related to selective attention. Evidence to support both assumptions is presented, followed by a description of a conditioned attention theory that emphasizes the role of initial levels of attention elicited by repeated relevant and irrelevant stimuli, and the differences between these levels in schizophrenia and normal groups. PMID- 15888433 TI - Various bilateral olfactory deficits in male patients with schizophrenia. AB - Olfactory identification deficits in schizophrenia patients are well documented. Less is known about the functioning of other olfactory domains and the possibility of lateralized dysfunctions. Thirty male schizophrenia patients and 30 male healthy controls underwent unirhinal assessment of various olfactory domains: detection threshold (dimethyl disulfide, phenyl ethanol), quality discrimination, and odor ratings (familiarity, pleasantness, edibility, intensity) of pure chemicals (Munich Olfaction Test), as well as familiarity and edibility judgments and identification of everyday odors. Aside from impaired identification, patients showed impaired familiarity and edibility judgments of everyday odors. With regard to odor ratings of pure chemicals, group differences were observed only in pleasantness ratings, with higher ratings in patients. Furthermore, patients had reduced sensitivity with dimethyl disulfide and reduced quality discrimination compared with controls. Further analyses showed that identification deficits were not attributable to reduced sensitivity but may be associated with impairments in quality discrimination. Olfactory dysfunctions were found across both nostrils. Results suggest specific dysfunctions in olfactory processing in schizophrenia patients, including early stages of the odor identification process. PMID- 15888434 TI - Symptoms versus neurocognitive test performance as predictors of psychosocial status in schizophrenia: a 1- and 4-year prospective study. AB - In recent years, a growing body of literature has highlighted the significance of neurocognitive deficits as markers for subsequent psychosocial deficits in patients with schizophrenia. Relatively few studies, however, have directly compared symptoms and neurocognitive test performance as predictors of psychosocial status in a prospective design. In two studies with schizophrenia patients, we investigated the relationship between symptom dimensions (psychomotor poverty, disorganization, reality distortion) and neurocognitive measures (problem solving, attention, verbal learning and memory) obtained at study entry, and psychosocial status measured at a 1- and 4-year followup. Results from the 1-year followup (n = 70) revealed that psychomotor poverty, symptoms of disorganization, and performance on measures of card-sorting and visual vigilance were related to psychosocial status. Results from the 4-year followup (n = 26) revealed a similar pattern of findings with the exception of verbal learning, which emerged as a predictor of psychosocial status only at the 4-year followup. Stepwise regression revealed that performance on measures of visual vigilance and psychomotor poverty symptoms explained the largest amount of variance in psychosocial status at both followup intervals. The significance of these findings for the development and assessment of novel treatment interventions for schizophrenia is discussed. PMID- 15888435 TI - Neurocognition, symptomatology, and functional skills in older alcohol-abusing schizophrenia patients. AB - Deficits in neurocognitive functioning are common to both schizophrenia and alcoholism. Recent studies suggest that neurocognitive functioning is the most significant predictor of social-adaptive functioning in schizophrenia. Cognitive impairment induced by alcoholism may result in more impaired functional outcome for comorbid patients. Past research examining alcohol-abusing schizophrenia patients has not examined correlates with functional outcome and has generally been limited to relatively younger patients. This study examined neurocognitive functioning and its correlates in alcohol-abusing schizophrenia patients between the ages of 40 and 80. Outpatients with schizophrenia (SZ; n = 17) or both schizophrenia and alcohol abuse or dependence (SZ + ETOH; n = 18) were tested on a neurocognitive battery, rated for symptomatology, and assessed for functional abilities. The results suggest that alcohol abuse in schizophrenia is associated with more impaired functioning across many domains, including memory impairment, negative and general psychopathology symptoms, and adaptive functions. The only significant predictor of impaired functional status in the overall sample and the SZ + ETOH group was neurocognitive functioning. PMID- 15888436 TI - Inhibition of cellular HIV-1 protease activity by lysyl-tRNA synthetase. AB - During early assembly of human immunodeficiency virus type 1 (HIV-1), an assembly complex is formed, the components of which include genomic RNA, Gag, GagPol, tRNA(Lys), and lysyl tRNA synthetase (LysRS). Directly increasing or decreasing cellular expression of LysRS results in corresponding changes in viral infectivity and in the viral concentrations of LysRS, tRNA(Lys), and, surprisingly, reverse transcriptase (RT). Since altering the cellular expression of LysRS does not lead to a change in the incorporation of the RT precursor protein, GagPol, in protease-negative HIV-1, we propose that the altered viral content of RT resulting from alterations in cellular LysRS concentration results from the ability of LysRS to inhibit premature activation of Gag-Pol viral protease within the complex. Supporting this hypothesis, we find that increases and decreases in cellular LysRS expression are accompanied by 5-8-fold increases and 5-fold decreases, respectively, in the cytoplasmic proteolysis of Gag and GagPol to mature viral proteins. Using a novel bioluminescence resonance energy transfer assay to directly measure HIV-1 protease activity in vivo also indicates that the overexpression of LysRS in the cell reduces viral protease activity. PMID- 15888437 TI - Dynamic interaction between the dual specificity phosphatase MKP7 and the JNK3 scaffold protein beta-arrestin 2. AB - JNK scaffold proteins bind JNK and upstream kinases to activate subsets of JNK and localize activated JNK to specific subcellular sites. We previously demonstrated that the dual specificity phosphatases (DSPs) MKP7 and M3/6 bind the scaffold JNK-interacting protein-1 (JIP-1) and inactivate the bound subset of JNK (1). The G protein-coupled receptor (GPCR) adaptor beta-arrestin 2 is also a JNK3 scaffold. It binds the upstream kinases ASK1 and MKK4 and couples stimulation of the angiotensin II receptor AT1aR to activation of a cytoplasmic pool of JNK3. Here we report that MKP7 also binds beta-arrestin 2 via amino acids 394-443 of MKP7, the same region that interacts with JIP-1. This region of MKP7 interacts with beta-arrestin 2 at a central region near the JNK binding domain. MKP7 dephosphorylates JNK3 bound to beta-arrestin 2, either following activation by ASK1 overexpression or following AT1aR stimulation. Initial AT1aR stimulation causes a rapid (within 5 min) dissociation of MKP7 from beta-arrestin 2. MKP7 then reassociates with beta-arrestin 2 on endocytic vesicles 30-60 min after initial receptor stimulation. This dynamic interaction between phosphatase and scaffold permits signal transduction through a module that binds both positive and negative regulators. PMID- 15888438 TI - Rhinoviruses infect human epithelial cells via ceramide-enriched membrane platforms. AB - The cell membrane contains very small distinct membrane domains enriched of sphingomyelin and cholesterol that are named rafts. We have shown that the formation of ceramide via activation of the acid sphingomyelinase transforms rafts into ceramide-enriched membrane platforms. These platforms are required for infection of mammalian cells with Pseudomonas aeruginosa, Staphylococcus aureus, or Neisseriae gonorrhoeae. In the present study we determined whether the acid sphingomyelinase, ceramide, and ceramide-enriched membrane platforms are also involved in the infection of human cells with pathogenic rhinoviruses. We demonstrate that infection of human epithelial cells with several rhinovirus strains triggers a rapid activation of the acid sphingomyelinase correlating with microtubules- and microfilament-mediated translocation of the enzyme from an intracellular compartment onto the extracellular leaflet of the cell membrane. The activity of the acid sphingomyelinase results in the formation of ceramide in the cell membrane and, finally, large ceramide-enriched membrane platforms. Rhinoviruses colocalize with ceramide-enriched membrane platforms during the infection. The significance of ceramide-enriched membrane platforms for rhinoviral uptake is demonstrated by the finding that genetic deficiency or pharmacological inhibition of the acid sphingomyelinase prevented infection of human epithelial cells by rhinoviruses. The data identify the acid sphingomyelinase and ceramide as key molecules for the infection of human cells with rhinoviruses. PMID- 15888439 TI - A double mutation in the extracellular Ca2+-sensing receptor's venus flytrap domain that selectively disables L-amino acid sensing. AB - The extracellular Ca(2+)-sensing receptor is activated allosterically by l-amino acids, and recent molecular analysis indicates that amino acids are likely to bind in the receptor's Venus flytrap domain. In the current study we set out to identify residues in the VFT domain that specifically support amino acid binding and/or amino acid-dependent receptor activation. Herein we describe two mutations of the Ca(2+)-sensing receptor (CaR) Venus Flytrap domain, T145A and S170T, that specifically impair amino acid sensing, leaving Ca2+ sensing intact, as determined by receptor-dependent activation of intracellular Ca2+ mobilization in fura-2-loaded HEK293 cells. With respect to the wild-type CaR, T145A and S170T exhibited reduced sensitivity to l-Phe, and T145A also exhibited markedly impaired l/d selectivity. When combined, the double mutant T145A/S170T exhibited normal or near-normal sensitivity to extracellular Ca2+ but was resistant to l Phe at concentrations up to 100 mm. We conclude that T145A/S170T selectively disables l-amino acid sensing and that the Ca2+ and l-amino acid-sensing functions of the CaR can be dissociated. PMID- 15888440 TI - Subunit assembly of N-methyl-d-aspartate receptors analyzed by fluorescence resonance energy transfer. AB - N-methyl-d-aspartate (NMDA) receptors play major roles in synaptic transmission and plasticity, as well as excitotoxicity. NMDA receptors are thought to be tetrameric complexes mainly composed of NMDA receptor (NR)1 and NR2 subunits. The NR1 subunits are required for the formation of functional NMDA receptor channels, whereas the NR2 subunits modify channel properties. Biochemical and functional studies indicate that subunits making up NMDA receptors are organized into a dimer of dimers, and the N termini of the subunits are major determinants for receptor assembling. Here we used a biophysical approach, fluorescence resonance energy transfer, to analyze the assembly of intact, functional NMDA receptors in living cells. The results showed that NR1, NR2A, and NR2B subunits could form homodimers when they were expressed alone in HEK293 cells. Subunit homodimers were also found existing in heteromeric NMDA receptors formed between NR1 and NR2 subunits. These findings are consistent with functional NMDA receptors being arranged as a dimer of dimers. In addition, our data indicated that the conformation of NR1 subunit homodimers was affected by the partner NR2 subunits during the formation of heteromeric receptor complexes, which might underlie the mechanism by which NR2 subunits modify NMDA receptor function. PMID- 15888441 TI - Base catalysis of chromophore formation in Arg96 and Glu222 variants of green fluorescent protein. AB - In green fluorescent protein (GFP), chromophore biosynthesis is initiated by a spontaneous main-chain condensation reaction. Nucleophilic addition of the Gly67 amide nitrogen to the Ser65 carbonyl carbon is catalyzed by the protein fold and leads to a heterocyclic intermediate. To investigate this mechanism, we substituted the highly conserved residues Arg96 and Glu222 in enhanced GFP (EGFP). In the R96M variant, the rate of chromophore formation is greatly reduced (time constant = 7.5 x 10(3) h, pH 7) and exhibits pH dependence. In the E222Q variant, the rate is also attenuated at physiological pH (32 h, pH 7) but is accelerated severalfold beyond that of EGFP at pH 9-10. In contrast, EGFP maturation is pH-independent and proceeds with a time constant of 1 h (pH 7-10). Mass spectrometric results for R96M and E222Q indicate accumulation of the pre cyclization state, consistent with rate-limiting backbone condensation. The pH rate profile implies that the Glu222 carboxylate titrates with an apparent pK(a) of 6.5 in R96M and that the Gly67 amide nitrogen titrates with an apparent pK(a) of 9.2 in E222Q. These data suggest a model for GFP chromophore synthesis in which the carboxylate of Glu222 plays the role of a general base, facilitating proton abstraction from the Gly67 amide nitrogen or the Tyr66 alpha-carbon. Arg96 fulfills the role of an electrophile by lowering the respective pK values and stabilizing the alpha-enolate. Modulating the base strength of the proton abstracting group may aid in the design of fast-maturing GFPs with improved characteristics for real-time monitoring of cellular events. PMID- 15888442 TI - Characterization of lysine 56 of histone H3 as an acetylation site in Saccharomyces cerevisiae. AB - Post-translational histone modifications abound and regulate multiple nuclear processes. Most modifications are targeted to the amino-terminal domains of histones. Here we report the identification and characterization of acetylation of lysine 56 within the core domain of histone H3. In the crystal structure of the nucleosome, lysine 56 contacts DNA. Phenotypic analysis suggests that lysine 56 is critical for histone function and that it modulates formamide resistance, ultraviolet radiation sensitivity, and sensitivity to hydroxyurea. We show that the acetylated form of histone H3 lysine 56 (H3-K56) is present during interphase, metaphase, and S phase. Finally, reverse genetic analysis indicates that none of the known histone acetyltransferases is solely responsible for H3 K56 acetylation in Saccharomyces cerevisiae. PMID- 15888443 TI - Cooperativity and pseudo-cooperativity in the glutathione S-transferase from Plasmodium falciparum. AB - Binding and catalytic properties of glutathione S-transferase from Plasmodium falciparum (PfGST) have been studied by means of fluorescence, steady state and pre-steady state kinetic experiments, and docking simulations. This enzyme displays a peculiar reversible low-high affinity transition, never observed in other GSTs, which involves the G-site and shifts the apparent K(D) for glutathione (GSH) from 200 to 0.18 mM. The transition toward the high affinity conformation is triggered by the simultaneous binding of two GSH molecules to the dimeric enzyme, and it is manifested as an uncorrected homotropic behavior, termed "pseudo-cooperativity." The high affinity enzyme is able to activate GSH, lowering its pK(a) value from 9.0 to 7.0, a behavior similar to that found in all known GSTs. Using 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, this enzyme reveals a potential optimized mechanism for the GSH conjugation but a low catalytic efficiency mainly due to a very low affinity for this co-substrate. Conversely, PfGST efficiently binds one molecule of hemin/monomer. The binding is highly cooperative (n(H) = 1.8) and occurs only when GSH is bound to the enzyme. The thiolate of GSH plays a crucial role in the intersubunit communication because no cooperativity is observed when S-methylglutathione replaces GSH. Docking simulations suggest that hemin binds to a pocket leaning into both the G-site and the H-site. The iron is coordinated by the amidic nitrogen of Asn-115, and the two carboxylate groups are in electrostatic interaction with the epsilon-amino group of Lys-15. Kinetic and structural data suggest that PfGST evolved by optimizing its binding property with the parasitotoxic hemin rather than its catalytic efficiency toward toxic electrophilic compounds. PMID- 15888444 TI - 7-Nitro-2,1,3-benzoxadiazole derivatives, a new class of suicide inhibitors for glutathione S-transferases. Mechanism of action of potential anticancer drugs. AB - Spectroscopic and rapid kinetic experiments were performed to detail the interaction of human glutathione S-transferases GSTA1-1, GSTM2-2, and GSTP1-1 with 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthio)hexanol (NBDHEX). This compound is a representative molecule of a new class of 7-nitro-2,1,3-benzoxadiazole (NBD) derivatives (non-GSH peptidomimetic compounds) that have been designed both to give strong GST inhibition and to accumulate in tumor cells avoiding the extrusion mechanisms mediated by the multidrug resistance protein pumps. We have recently shown that submicromolar amounts of NBDHEX trigger apoptosis in several human tumor cell lines through the dissociation of the JNK.GSTP1-1 complex (Turella, P., Cerella, C., Filomeni, G., Bullo, A., De Maria, F., Ghibelli, L., Ciriolo, M. R., Cianfriglia, M., Mattei, M., Federici, G., Ricci, G., and Caccuri, A. M. (2005) Cancer Res. 65, 3751-3761). Results reported in the present study indicated that NBDHEX behaves like a suicide inhibitor for GSTs. It bound to the H-site and was conjugated with GSH forming a sigma complex at the C-4 of the benzoxadiazole ring. This complex was tightly stabilized in the active site of GSTP1-1 and GSTM2-2, whereas in GSTA1-1 the release of the 6-mercapto-1 hexanol from the sigma complex was the favored event. Docking studies demonstrated the likely localization of the sigma complex in the GST active sites and provide a structural explanation for its strong stabilization. PMID- 15888445 TI - 5-Formyltetrahydrofolate is an inhibitory but well tolerated metabolite in Arabidopsis leaves. AB - 5-Formyltetrahydrofolate (5-CHO-THF) is formed via a second catalytic activity of serine hydroxymethyltransferase (SHMT) and strongly inhibits SHMT and other folate-dependent enzymes in vitro. The only enzyme known to metabolize 5-CHO-THF is 5-CHO-THF cycloligase (5-FCL), which catalyzes its conversion to 5,10 methenyltetrahydrofolate. Because 5-FCL is mitochondrial in plants and mitochondrial SHMT is central to photorespiration, we examined the impact of an insertional mutation in the Arabidopsis 5-FCL gene (At5g13050) under photorespiratory (30 and 370 micromol of CO2 mol(-1)) and non-photorespiratory (3200 micromol of CO2 mol(-1)) conditions. The mutation had only mild visible effects at 370 micromol of CO2 mol(-1), reducing growth rate by approximately 20% and delaying flowering by 1 week. However, the mutation doubled leaf 5-CHO-THF level under all conditions and, under photorespiratory conditions, quadrupled the pool of 10-formyl-/5,10-methenyltetrahydrofolates (which could not be distinguished analytically). At 370 micromol of CO2 mol(-1), the mitochondrial 5 CHO-THF pool was 8-fold larger in the mutant and contained most of the 5-CHO-THF in the leaf. In contrast, the buildup of 10-formyl-/5,10 methenyltetrahydrofolates was extramitochondrial. In photorespiratory conditions, leaf glycine levels were up to 46-fold higher in the mutant than in the wild type. Furthermore, when leaves were supplied with 5-CHO-THF, glycine accumulated in both wild type and mutant. These data establish that 5-CHO-THF can inhibit SHMT in vivo and thereby influence glycine pool size. However, the near-normal growth of the mutant shows that even exceptionally high 5-CHO-THF levels do not much affect fluxes through SHMT or any other folate-dependent reaction, i.e. that 5-CHO-THF is well tolerated in plants. PMID- 15888446 TI - Mutations in TFIIIA that increase stability of the TFIIIA-5 S rRNA gene complex: unusual effects on the kinetics of complex assembly and dissociation. AB - We have identified four mutations in Xenopus TFIIIA that increase the stability of TFIIIA-5 S rRNA gene complexes. In each case, the mutation has a relatively modest effect on equilibrium binding affinity. In three cases, these equilibrium binding effects can be ascribed primarily to decreases in the rate constant for protein-DNA complex dissociation. In the fourth case, however, a substitution of phenylalanine for the wild-type leucine at position 148 in TFIIIA results in much larger compensating changes in the kinetics of complex assembly and dissociation. The data support a model in which a relatively unstable population of complexes with multi-component dissociation kinetics forms rapidly; complexes then undergo a slow conformational change that results in very stable, kinetically homogeneous TFIIIA-DNA complexes. The L148F mutant protein acts as a particularly potent transcriptional activator when it is fused to the VP16 activation domain and expressed in yeast cells. Substitution of L148 to tyrosine or tryptophan produces an equally strong transcriptional activator. Substitution to histidine results in genetic and biochemical effects that are more modest than, but similar to, those observed with the L148F mutation. We propose that an amino acid with a planar side chain at position 148 can intercalate between adjacent base pairs in the intermediate element of the 5 S rRNA gene. Intercalation occurs slowly but results in a very stable DNA-protein complex. These results suggest that transcriptional activation by a cis-acting sequence element is largely dependent on the kinetic, rather than the thermodynamic, stability of the complex formed with an activator protein. Thus, transcriptional activation is dependent in large part on the lifetime of the activator-DNA complex rather than on binding site occupancy at steady state. Introduction of intercalating amino acids into zinc finger proteins may be a useful tool for producing artificial transcription factors with particularly high in vivo activity. PMID- 15888447 TI - cAMP-response elements in Aplysia creb1, creb2, and Ap-uch promoters: implications for feedback loops modulating long term memory. AB - The Aplysia genes encoding for cAMP-response element-binding protein 1 (CREB1), CREB2, and ubiquitin C-terminal hydrolase (Ap-uch) have been implicated in the formation of long term memory. However, nothing is known about the promoter regions of these genes or the transcription factors that regulate them. We cloned the promoter regions of creb1, creb2, and Ap-uch and identified a canonical cAMP response element (CRE) in the promoter region of creb1. Variants of the canonical CRE were identified in all three promoters. TATA boxes and C/EBP-binding motifs are also present in the promoter regions of these genes. Promoter immunoprecipitation assays and chromatin immunoprecipitation assays indicated that CREB1 and CREB2 bind to the promoter regions of creb1 and creb2, suggesting that feedback loops modulate the formation of long term memory. In a positive feedback loop, phosphorylated CREB1 might induce its own gene via CREs. In support of this suggestion, treatment with serotonin enhanced binding of CREB1 to its promoter region and increased mRNA levels of creb1. Levels of Ap-uch mRNA also increased in response to serotonin; however, binding of CREB1 or CREB2 to the promoter region of Ap-uch was not detected. The finding that the promoter region of creb2 has a CRE raises the intriguing possibility that its expression is regulated by CREB1 and/or CREB2. CREB2 may repress its own gene, forming a negative feedback loop, and CREB2 up-regulation via CREB1 may limit the activity of the CREB1-mediated positive feedback loop. PMID- 15888448 TI - The low density lipoprotein receptor regulates the level of central nervous system human and murine apolipoprotein E but does not modify amyloid plaque pathology in PDAPP mice. AB - Apolipoprotein E (apoE), a chaperone for the amyloid beta (Abeta) peptide, regulates the deposition and structure of Abeta that deposits in the brain in Alzheimer disease (AD). The primary apoE receptor that regulates levels of apoE in the brain is unknown. We report that the low density lipoprotein receptor (LDLR) regulates the cellular uptake and central nervous system levels of astrocyte-derived apoE. Cells lacking LDLR were unable to appreciably endocytose astrocyte-secreted apoE-containing lipoprotein particles. Moreover, cells overexpressing LDLR showed a dramatic increase in apoE endocytosis and degradation. We also found that LDLR knock-out (Ldlr-/-) mice had a significant, approximately 50% increase in the level of apoE in the cerebrospinal fluid and extracellular pools of the brain. However, when the PDAPP mouse model of AD was bred onto an Ldlr-/- background, we did not observe a significant change in brain Abeta levels either before or after the onset of Abeta deposition. Interestingly, human APOE3 or APOE4 (but not APOE2) knock-in mice bred on an Ldlr-/- background had a 210% and 380% increase, respectively, in the level of apoE in cerebrospinal fluid. These results demonstrate that central nervous system levels of both human and murine apoE are directly regulated by LDLR. Although the increase in murine apoE caused by LDLR deficiency was not sufficient to affect Abeta levels or deposition by 10 months of age in PDAPP mice, it remains a possibility that the increase in human apoE3 and apoE4 levels caused by LDLR deficiency will affect this process and could hold promise for therapeutic targets in AD. PMID- 15888449 TI - Ubiquitin-proteasome-mediated degradation, intracellular localization, and protein synthesis of MyoD and Id1 during muscle differentiation. AB - Mammalian skeletal myogenesis results in the differentiation of myoblasts to mature syncytial myotubes, a process regulated by an intricate genetic network of at least three protein families: muscle regulatory factors, E proteins, and Id proteins. MyoD, a key muscle regulatory factor, and its negative regulator Id1 have both been shown to be degraded by the ubiquitin-proteasome system. Using C2C12 cells and confocal fluorescence microscopy, we showed that MyoD and Id1 co localize within the nucleus in proliferating myoblasts. In mature myotubes, in contrast, they reside in distinctive subcellular compartments, with MyoD within the nucleus and Id1 exclusively in the cytoplasm. Cellular abundance of Id1 was markedly diminished from the very onset of muscle differentiation, whereas MyoD abundance was reduced to a much lesser extent and only at the later stages of differentiation. These reductions in MyoD and Id1 protein levels seem to result from a change in the rate of protein synthesis rather than the rate of degradation. In vivo protein stability studies revealed that the rates of ubiquitin-proteasome-mediated MyoD and Id1 degradation are independent of myogenic differentiation state. Id1 and MyoD were both rapidly degraded, each with a t 1/2 approximately = 1 h in myoblasts and in myotubes. Furthermore, relative protein synthesis rates for MyoD and Id1 were significantly diminished during myoblast to myotube differentiation. These results provide insight as to the interaction between MyoD and Id1 in the process of muscle differentiation and have implications for the involvement of the ubiquitin-proteasome-mediated protein degradation and protein synthesis in muscle differentiation and metabolism under abnormal and pathological conditions. PMID- 15888450 TI - Novelty of the pyruvate metabolic enzyme dihydrolipoamide dehydrogenase in spermatozoa: correlation of its localization, tyrosine phosphorylation, and activity during sperm capacitation. AB - Spermatozoa are cells distinctly different from other somatic cells of the body, capacitation being one of the unique phenomena manifested by this gamete. We have shown earlier that dihydrolipoamide dehydrogenase, a post-pyruvate metabolic enzyme, undergoes capacitation-dependent tyrosine phosphorylation, and the functioning of the enzyme is required for hyperactivation (enhanced motility) and acrosome reaction of hamster spermatozoa (Mitra, K., and Shivaji, S. (2004) Biol. Reprod. 70, 887-899). In this report we have investigated the localization of this mitochondrial enzyme in spermatozoa revealing non-canonical extra mitochondrial localization of the enzyme in mammalian spermatozoa. In hamster spermatozoa, dihydrolipoamide dehydrogenase along with its host complex, the pyruvate dehydrogenase complex, are localized in the acrosome and in the principal piece of the sperm flagella. The localization of dihydrolipoamide dehydrogenase, however, appears to be in the mitochondria in the spermatocytes, but in spermatids it appears to show a juxtanuclear localization (like Golgi). The capacitation-dependent time course of tyrosine phosphorylation of dihydrolipoamide dehydrogenase appears to be different in the principal piece of the flagella and the acrosome in hamster spermatozoa. Activity assays of this bi directional enzyme suggest a strong correlation between the tyrosine phosphorylation and the bi-directional enzyme activity. This is the first report of a direct correlation of the localization, tyrosine phosphorylation, and activity of the important metabolic enzyme, dihydrolipoamide dehydrogenase, implicating dual involvement and regulation of the enzyme during sperm capacitation. PMID- 15888451 TI - Reorganization of ErbB family and cell survival signaling after Knock-down of ErbB2 in colon cancer cells. AB - The role of the ErbB family in supporting the malignant phenotype was characterized by stable transfection of a single chain antibody (ScFv5R) against ErbB2 containing a KDEL endoplasmic reticulum retention sequence into GEO human colon carcinoma cells. The antibody traps ErbB2 in the endoplasmic reticulum, thereby down-regulating cell surface ErbB2. The transfected cells showed inactivation of ErbB2 tyrosine phosphorylation and reduced heterodimerization of ErbB2 and ErbB3. This resulted in greater sensitivity to apoptosis induced by growth deprivation and delayed tumorigenicity in vivo. Furthermore, decreased heterodimerization of ErbB2 and ErbB3 led to a reorganization in ErbB function in transfected cells as heterodimerization between epidermal growth factor receptor (EGFR) and ErbB3 increased, whereas ErbB3 activation remained almost the same. Importantly, elimination of ErbB2 signaling resulted in an increase in EGFR expression and activation in transfected cells. Increased EGFR activation contributed to the sustained cell survival in transfected cells. PMID- 15888452 TI - Cdc2-mediated inhibition of epidermal growth factor activation of the extracellular signal-regulated kinase pathway during mitosis. AB - Inhibition of general transcription and translation occurs during mitosis to preserve the high energy requirements needed for the dynamic structural changes that are occurring at this time of the cell cycle. Although the mitotic kinase Cdc2 appears to directly phosphorylate and inhibit key proteins directly involved in transcription and translation, the role of Cdc2 in regulating up-stream growth factor receptor-mediated signal transduction pathways is limited. In the present study, we examined mechanisms involved in uncoupling receptor-mediated activation of the extracellular signal-regulated (ERK) signaling pathway in mitotic cells. Treatment with epidermal growth factor (EGF) failed to activate the ERK pathway in mitotic cells, although partial activation of ERK could be achieved in mitotic cells treated with phorbol 12-myristate 13-acetate (PMA). The discrepancy between EGF and PMA-mediated ERK activation suggested that multiple events in the ERK pathway were regulated during mitosis. We show that Cdc2 inhibits EGF-mediated ERK activation through direct interaction and phosphorylation of several ERK pathway proteins, including the guanine nucleotide exchange factor, Sos-1, and Raf-1 kinase. Inhibition of Cdc2 activity with roscovitine in mitotic cells restored ERK activation by EGF and PMA. Similarly, mitotic inhibition of ERK activity in cells expressing active mutants of H-Ras and Raf-1 kinase could also be reversed following Cdc2 inhibition. In contrast, ERK activation in cells expressing active MEK1 was not inhibited during mitosis or affected by roscovitine. These data suggest that Cdc2 inhibits growth factor receptor mediated ERK activation during mitosis by primarily targeting signaling proteins that are upstream of MEK1. PMID- 15888453 TI - Physiologic and pharmacologic factors influencing glyceroneogenic contribution to triacylglyceride glycerol measured by mass isotopomer distribution analysis. AB - An imbalance between triacylglycerol synthesis and breakdown is necessary for the development of obesity. The direct precursor for triacylglycerol biosynthesis is alpha-glycerol phosphate, which can have glycolytic and glyceroneogenic origins. We present a technique for determining the relative glyceroneogenic contribution to triacylglyceride glycerol by labeling the glycerol moiety with 2H2O. The number of hydrogen atoms (n) incorporated from H2O into C-H bonds reflects the metabolic source of alpha-glycerol phosphate and can be calculated by combinatorial analysis of the distribution of mass isotopomers in triacylglyceride glycerol. Three physiological settings with potential effects on glyceroneogenesis and glycolysis were studied in rodents. Adipose tissue acylglyceride glycerol in mice fed a low carbohydrate diet had significantly higher values of n than in mice fed a high carbohydrate diet, suggesting an increased contribution from glyceroneogenesis of from 17 to 50% on the low carbohydrate diet. Similarly, mice administered rosiglitazone had a significant relative increase in glyceroneogenesis (from 17 to 53%), indicated by an increase in adipose acylglyceride glycerol n. Fructose infusion in overnight fasted rats rapidly lowered plasma triacylglyceride glycerol n, reflecting a decreased contribution from glyceroneogenesis (from 66 to 34%) presumably because of increased glycolytic input. In conclusion, we demonstrate that the number of C-H atoms derived from cellular H2O in triacylglyceride glycerol is an informative indicator of alpha-glycerol phosphate origin and, ultimately, triacylglycerol metabolism. Under certain physiological conditions, glyceroneogenesis can be up regulated in adipose (e.g. low carbohydrate diet) or down-regulated in liver (e.g. fructose infusion). Additionally, stimulation of glyceroneogenesis by rosiglitazone in adipose tissue may be an important factor in the antilipolytic actions of thiazolidinediones. PMID- 15888454 TI - Double-spanning plant viral movement protein integration into the endoplasmic reticulum membrane is signal recognition particle-dependent, translocon-mediated, and concerted. AB - The current model for cell-to-cell movement of plant viruses holds that transport requires virus-encoded movement proteins that intimately associate with endoplasmic reticulum membranes. We have examined the early stages of the integration into endoplasmic reticulum membranes of a double-spanning viral movement protein using photocross-linking. We have discovered that this process is cotranslational and proceeds in a signal recognition particle-dependent manner. In addition, nascent chain photocross-linking to Sec61alpha and translocating chain-associated membrane protein reveal that viral membrane protein insertion takes place via the translocon, as with most eukaryotic membrane proteins, but that the two transmembrane segments of the viral protein leave the translocon and enter the lipid bilayer together. PMID- 15888455 TI - Glutathionylation of two cysteine residues in paired domain regulates DNA binding activity of Pax-8. AB - We reported that the first two cysteine residues out of three present in paired domain (PD), a DNA-binding domain, are responsible for redox regulation of Pax-8 DNA binding activity. We show that glutathionylation of these cysteines has a regulatory role in PD binding. Wild-type PD and its mutants with substitution of cysteine to serine were synthesized and named CCC, CSS, SCS, SSC, and SSS according to the positions of substituted cysteines. They were incubated in a buffer containing various ratios of GSH/GSSG and subjected to gel shift assay. Binding of CCC, CSS, and SCS was impaired with decreasing GSH/GSSG ratio, whereas that of SSC and SSS was not affected. Because [3H]glutathione was incorporated into CCC, CSS, and SCS, but not into SSC and SSS, the binding impairment was ascribed to glutathionylation of the redox-reactive cysteines. This oxidative inactivation of PD binding was reversed by a reductant dithiothreitol and by redox factor (Ref)-1 in vitro. To explore the glutathionylation in cells, Chinese hamster ovary cells overexpressing CSS and SCS were labeled with [35S]cysteine in the presence of cycloheximide. Immunoprecipitation with an antibody against PD revealed that treatment of the cells with an oxidant diamide induced the 35S incorporation into both mutants, suggesting the PD glutathionylation in cells. Since the two cysteine residues in PD are conserved in all Pax members, this novel posttranslational modification of PD would provide a new insight into molecular basis for modulation of Pax function. PMID- 15888456 TI - Structural determinants of the agonist-independent association of human peroxisome proliferator-activated receptors with coactivators. AB - Lipid homeostasis is controlled by various nuclear receptors (NRs), including the peroxisome proliferator-activated receptors (PPARalpha, delta, and gamma), which sense lipid levels and regulate their metabolism. Here we demonstrate that human PPARs have a high basal activity and show ligand-independent coactivator (CoA) association comparable with the NR constitutive androstane receptor. Using PPARgamma as an example, we found that four different amino acid groups contribute to the ligand-independent stabilization of helix 12 of the PPAR ligand binding domain. These are: (i) Lys329 and Glu499, mediating a charge clamp-type stabilization of helix 12 via a CoA bridge; (ii) Glu352, Arg425, and Tyr505, directly stabilizing the helix via salt bridges and hydrogen bonds; (iii) Lys347 and Asp503, interacting with each other as well as contacting the CoA; and (iv) His351, Tyr(355), His477, and Tyr501, forming a hydrogen bond network. These amino acids are highly conserved within the PPAR subfamily, suggesting that the same mechanism may apply for all three PPARs. Phylogenetic trees of helix 12 amino acid and nucleotide sequences of all crystallized NRs and all human NRs, respectively, indicated a close relationship of PPARs with constitutive androstane receptor and other constitutive active members of the NR superfamily. Taking together, the ligand-independent tight control of the position of the PPAR helix 12 provides an effective alternative for establishing an interaction with CoA proteins. This leads to high basal activity of PPARs and provides an additional view on PPAR signaling. PMID- 15888457 TI - Role of Pseudomonas aeruginosa quorum-sensing systems in a mouse model of chronic respiratory infection. AB - The role of quorum-sensing systems in a mouse model of chronic Pseudomonas aeruginosa infection was studied. A chronic P. aeruginosa respiratory infection model was established by placement of a tube pre-coated with strain PAO1 (wild type) or a quorum-sensing mutant, namely PAO-JP1 (Delta lasI), PDO100 (Delta rhlI) or PAO-JP2 (Delta lasI/Delta rhlI), in the bronchus. At day 14 after infection, the numbers of viable bacteria in the quorum-sensing-mutant groups were lower than in the wild-type group. Histopathological examination showed milder inflammatory changes in the lungs infected with the mutant groups compared with the wild-type group. In the bronchoalveolar lavage fluid from the quorum sensing-system-mutant groups the proportion of neutrophils was lower than in wild type group. These findings indicate that the quorum-sensing system plays an important role in chronic P. aeruginosa respiratory infection. PMID- 15888458 TI - Genotypic and phenotypic characteristics of Pseudomonas aeruginosa isolates associated with ulcerative keratitis. AB - A collection of 63 isolates of Pseudomonas aeruginosa associated with ulcerative keratitis, collected from six centres in England, were typed using serotyping and random amplified polymorphic DNA-PCR, and screened for several variable virulence related genotypes and phenotypes. Sixty-one percent of the isolates were of either serotype O1 or serotype O11, but there was no evidence for a common clone. The majority of isolates (59%) were PCR-positive for exoU rather than for exoS (38%), and carried a-type fliC genes (76%) rather than b-type (24%). Isolates were PCR-positive for pyoverdine-receptor types at a prevalence of 38% for type I, 46 % for type II and 8 % for type III. All but one of the isolates exhibited twitching activity. There was a correlation between the presence of exoS and twitching activity (P = 0.04), suggesting that a combination of exoS genotype and good twitching activity may have a role to play in ExoU-independent corneal virulence. PMID- 15888459 TI - Reduced interleukin-18 secretion in Brucella abortus 2308-infected murine peritoneal macrophages and in spleen cells obtained from B. abortus 2308-infected mice. AB - Th1 immune responses in which gamma interferon (IFN-gamma) production predominates are associated with protective immunity against intracellular bacteria. Following infection, interleukin-18 (IL-18) may contribute, in association with IL-12, to optimal IFN-gamma production. In this study, the secretion of IL-18 following intracellular infection with virulent Brucella abortus 2308 in CD-1 cultured peritoneal macrophages and splenocyte cultures was investigated. The production of IL-18 was reduced in both CD-1 mouse peritoneal macrophages infected with B. abortus 2308 and splenocyte cultures obtained from B. abortus 2308-infected mice at 3, 6 and 10 days post-infection (p.i.). In contrast, splenocyte cultures obtained from B. abortus 2308-infected mice at 3 days p.i. secreted significant amounts of IFN-gamma. Stimulation of these cells with recombinant IL-18 (rIL-18) and/or rIL-12 did not significantly increase IFN gamma secretion at the splenocyte level. These data suggest that once the infection has been established, B. abortus 2308 selectively limits IL-18 secretion without affecting endogenous IFN-gamma production. PMID- 15888460 TI - Correlation of antigenic expression with progress in antibiotic therapy of acute human brucellosis. AB - Human brucellosis is a zoonotic disease which is endemic in Saudi Arabia. The aim of this study was to investigate the humoral immune responses and identify the target antigens that persist at different stages in human brucellosis during antibiotic therapy. To do this, an acute case of accidental nosocomial infection was studied experimentally. Blood was collected from the patient at the time of diagnosis, and at weekly intervals during therapy until remission. IgG and IgM immunoblotting was used to characterize specific antigenic determinants, and ELISA antibody titration was performed to quantify the circulating antibodies. Results indicated that protein bands of 12-13.5 kDa bound IgG in the patient's sera but did not bind IgM on immunoblots and are probably not specific for, or important in, early stage infections. However, an 18 kDa band persisted during infection through remission. The pivotal and most important findings were that the number of protein bands seen on immunoblots, the magnitude of ELISA antibody titres and the concomitant changes in the intensity of the polypeptide bands of 42-43 kDa were positively correlated with the stage of infection. High numbers of anti-IgG and -IgM immunoblot bands coupled with high ELISA antibody titres and a concomitant increase in intensity of the 42-43 kDa bands were positively correlated with acute and severe infection. Conversely, a reduction in the number of polypeptide bands as well as a decrease in the intensity, until the complete disappearance of the 42-43 kDa bands, coupled with low (baseline) ELISA antibody titration values indicated successful treatment and remission. The routine use of the methods described here to ascertain the stage of the disease, assess the progress of antimicrobial therapy and monitor cases of relapse in human brucellosis is suggested. PMID- 15888461 TI - Diagnosis of partially treated culture-negative bacterial meningitis using 16S rRNA universal primers and restriction endonuclease digestion. AB - Cerebrospinal fluid (CSF) obtained from patients with partially treated and culture-negative meningitis was subjected to PCR using 16S rDNA universal primers followed by restriction endonuclease digestion. In all, 43 patients and 7 controls were enrolled in this study. Twenty-one meningitic samples were positive by PCR. Mycobacterium tuberculosis was the causative agent in seven cases followed by Haemophilus influenzae (four), Streptococcus pneumoniae (two), Listeria monocytogenes (one), Escherichia coli (one), Pseudomonas aeruginosa (one) and Staphylococcus aureus (one). Only two meningitic CSF samples were culture-positive. In this study, PCR using bacterial 16S rDNA specific universal primers was found to be superior to conventional methods in the diagnosis of partially treated meningitis. PMID- 15888462 TI - Subtyping of Clostridium difficile PCR ribotype 001 by REP-PCR and PFGE. AB - The REP-PCR (repetitive sequence-based PCR using repetitive extragenic palindromic primers) typing method and a modified PFGE method were applied to isolates of Clostridium difficile PCR ribotype 001 with the aim of comparing their performance as methods of subtyping this organism. Of 200 isolates from 60 hospitals tested by REP-PCR, eight subtypes were identified and labelled as REP PCR subtypes 001-008. The predominant subtype, REP-PCR subtype 003, accounted for 47% of the total. Fifty-two of the 200 isolates were analysed by a modified PFGE method and seven subtypes were identified, labelled as PF-A-PF-G. There was excellent correlation between REP-PCR subtypes and PFGE subtypes with both methods displaying broadly similar discriminatory powers. However, REP-PCR subtyping proved to be a much easier, cheaper and more rapid method suitable for application for routine subtyping of C. difficile ribotype 001. Application of REP-PCR subtyping to UK isolates of C. difficile PCR ribotype 001 from 60 different centres revealed a wide distribution of REP-PCR subtype 003 throughout England and Wales, with a regional clustering of REP-PCR subtype 001 around Northwest England and North Wales. Analysis of isolates from a single hospital over a 4-year period revealed a change in predominant subtype over time. PMID- 15888463 TI - In vitro antifungal activity of 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1 methylethyl pentanoate, a dihydropyrrole derivative. AB - A novel compound 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate was isolated from the plant Datura metel L. The in vitro activity of this dihydropyrrole derivative against Aspergillus and Candida species was evaluated by using standard methods approved by the National Committee for Clinical Laboratory Standards. The compound was found to be active against all the species tested, namely Candida albicans, Candida tropicalis, Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. The MIC at which more than 90% of growth was inhibited (MIC(90)) by the compound ranged from 21.87 to 43.75 microg ml(-1) against various fungal species by microbroth dilution assay. Since the compound 2-(3,4-dimethyl-2,5-dihydro-1H-pyrrol-2-yl)-1-methylethyl pentanoate has antifungal activity it can be explored further to develop new antimycotic drugs. PMID- 15888464 TI - Long-term effects on the nasopharyngeal flora of children following antimicrobial therapy of acute otitis media with cefdinir or amoxycillin-clavulanate. AB - The effect on the nasopharyngeal bacterial flora of 10 days of amoxycillin clavulanate or cefdinir antimicrobial therapy was studied in 50 children with acute otitis media. Before therapy, 17 potential pathogens (Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis) were isolated from the nasopharynx of 14 (56%) of those treated with amoxycillin-clavulanate, and 20 potential pathogens were recovered from 15 (60%) of those treated with cefdinir. Following therapy, at days 12-15, the number of potential pathogens was reduced to a similar extent with both therapies, to three in those treated with amoxycillin-clavulanate and two in those treated with cefdinir. However, the number of potential pathogens rebounded faster in those treated with amoxycillin clavulanate as compared with cefdinir in the two subsequent specimens taken at days 30-35 and 60-65 (12 and 18 in the amoxycillin-clavulanate group, and six and nine in the cefdinir group, P < 0.01 and P < 0.001, respectively). Differences between the groups were also noted in the recovery of organisms with interfering capability. Immediately following amoxycillin-clavulanate therapy, the number of interfering organisms declined from 54 to 13, while following cefdinir treatment their number was reduced from 59 to 39 (P < 0.001). The differences between the two therapy groups persisted in the two later specimens taken at days 30-35 and 60-65 (25 and 38 in the amoxycillin-clavulanate group, and 52 and 51 in the cefdinir group, P < 0.001 and P < 0.05, respectively). This study illustrates the potential beneficial effect of using a narrow-spectrum antimicrobial that selectively spares the interfering organisms while eliminating pathogens. The benefit of such therapy is the prevention of reacquisition of pathogenic bacteria in the nasopharynx. In contrast, utilization of a broad-spectrum antimicrobial is associated with prolonged absence of inhibitory organisms and rapid recolonization with pathogens. PMID- 15888465 TI - Susceptibilities to antiseptic agents and distribution of antiseptic-resistance genes qacA/B and smr of methicillin-resistant Staphylococcus aureus isolated in Asia during 1998 and 1999. AB - Many antiseptic agents are used in hygienic handwashes in the prevention of nosocomial infections by methicillin-resistant Staphylococcus aureus (MRSA). The plasmid-borne genes qacA/B and smr confer resistance to cationic antiseptic agents in S. aureus. In this study, the susceptibilities for dyes and antiseptic agents (e.g. acriflavine, acrinol, benzalkonium chloride, benzethonium chloride, chlorhexidine digluconate and alkyldiaminoethylglycine hydrochloride) of 894 isolates of MRSA collected from 11 Asian countries (South Korea, China, the Philippines, Singapore, Vietnam, Thailand, Indonesia, India, Sri Lanka, Saudi Arabia and Japan) between 1998 and 1999 were examined. In addition, the distributions of the antiseptic-resistance genes qacA/B and smr were studied by PCR. Among the Asian MRSA isolates 57.7% (516/894) were acriflavine-resistant. The smr gene was detected in 31.6% (12/38) of MRSA isolates from India but only in 1.9% (16/856) of all the isolates from other Asian countries. MRSA with qacA/B comprised 41.6% (372/894) of the isolates across Asia. In addition, PFGE was performed to type the MRSA and grouped the tested 30 MRSA isolates with qacA/B into 21 PFGE types. The results indicated that qacA/B is functionally the most important gene mediating antiseptic resistance in the MRSA strains of Asia and that a specific MRSA with qacA/B was not prevalent in Asia but qacA/B were widely spread among MRSA of Asia, while the geographical distribution of smr is more limited. PMID- 15888466 TI - Helicobacter pylori antibiotic-resistance patterns and risk factors in adult dyspeptic patients from ethnically diverse populations in central and south London during 2000. AB - Surveillance of Helicobacter pylori antibiotic susceptibility from patients in London, the largest metropolitan area in the UK, is limited, despite resistance being a key factor in treatment failure. A two-centre survey was performed over 12 months (1999-2000) to determine antibiotic-resistance rates of isolates from dyspeptic patients attending endoscopy clinics serving two ethnically diverse central and south London communities. The in vitro antibiotic susceptibilities were determined from disc diffusion and epsilometer (E) tests on 101 H. pylori isolates. Overall resistance rates were 59% for metronidazole and 11% for clarithromycin, with 8 % resistance to both antibiotics. Corresponding primary resistance rates were 50% and 7%, respectively. High-level-resistance was a feature of 82% of the metronidazole (MIC > or = 256 mg l(-1)) -resistant and 55% of the clarithromycin (MIC > or = 32 mg l(-1)) -resistant strains. All isolates were susceptible to amoxycillin and tetracycline. No associations between resistance and either the gender or the age of the patients were detected. The main risk for resistance to metronidazole was non-UK birth as comparative rates were 68% for non-UK vs. 40% for UK-born individuals. Resistant isolates were genotypically diverse with respect to cagA/vacA type. Four 23S rDNA nucleotide polymorphisms were associated with clarithromycin resistance, mostly (9/11) at A2143G. In conclusion, the high overall metronidazole-resistance rate of 59% for H. pylori from inner London was twice the rate found in other UK-based studies and was attributed to the higher risk of resistant strains infecting individuals born outside the UK. The need for continued resistance surveillance is indicated to monitor the effects of the 'test and treat' strategy for H. pylori eradication, particularly of isolates from at-risk individuals. PMID- 15888467 TI - Origins and properties of Mycobacterium tuberculosis isolates in London. AB - Using similarities of IS6110 banding patterns, isolates of Mycobacterium tuberculosis from a population-based study in London were assigned to 12 large groups termed 'superfamilies' (sfams). Analysis of patient data showed a marked geographical association in the distribution of these sfams. In particular, isolates from patients born in Europe were from different sfams than those born elsewhere, indicating that there had been relatively little transmission of tuberculosis in London from immigrant communities into the endogenous population. Multivariate analysis showed that certain sfams were significantly associated with pulmonary rather than extrapulmonary disease, or with sputum smear negativity, independently of country of birth or ethnicity, suggesting that the properties of the infecting organism play a role in the nature of the disease process. PMID- 15888468 TI - Phospholipase B enzyme expression is not associated with other virulence attributes in Candida albicans isolates from patients with human immunodeficiency virus infection. AB - The extracellular phospholipases of Candida albicans are considered to play a significant role in the pathogenesis of human infections. Therefore 30 clinical isolates of C. albicans from human immunodeficiency virus (HIV)-infected individuals were screened for phospholipase production in vitro (using an egg yolk-agar medium). Two groups of six isolates with positive (group A) or deficient (group B) phospholipase activity were then analysed for phospholipase B1 (PLB1) gene expression both in egg-yolk-agar and yeast extract/peptone/dextrose (YPD) broth media. A total of four virulence attributes of these two groups were in turn characterized, namely their germ-tube formation, cell-surface-hydrophobicity (CSH), adhesion to buccal epithelial cells (ABEC) and haemolysin production, and these factors were subsequently correlated with PLB1 expression. In the phospholipase-producing isolates (group A) a positive correlation was demonstrated between phospholipase production and the degree of PLB1 expression in YPD medium (r = 0.96, P < 0.01). No such association was observed in group A isolates for PLB1 expression in egg-yolk-agar medium. Further, PLB1 expression in egg-yolk agar was less than that in YPD medium, although a positive correlation was seen between the expression levels on regression analysis (r = 0.86, P = 0.026). Surprisingly, however, no significant associations were observed in either growth media between PLB1 expression and any of the four pathogenic attributes examined (P < 0.001). A significant correlation was seen between CSH and ABEC (r = 0.74) in group A isolates. The phospholipase deficient group B, however, demonstrated a significant correlation between the latter parameters (r = +0.50) and also between germ-tube formation and ABEC (r = 0.59), and germ-tube formation and haemolysin production (r = +0.31). It appears that in oral C. albicans isolates in HIV infection there may be no significant association between the degree of PLB1 expression and other widely recognized major virulence attributes. PMID- 15888469 TI - Bacteriology of chronic maxillary sinusitis associated with nasal polyposis. AB - Aspirates from 48 chronically inflamed maxillary sinuses from patients who had nasal polyposis were processed for aerobic and anaerobic bacteria. Bacterial growth was present in 46 (96%) specimens. Aerobic or facultative bacteria were present in 6 (13%) specimens, anaerobic bacteria alone in 18 (39%), and mixed aerobic and anaerobic bacteria in 22 (48%). There were 110 bacterial isolates (2.4 per specimen). Thirty-nine of the isolates were aerobic or facultative organisms (0.85 per specimen). The predominant aerobic or facultative organisms were: Staphylococcus aureus, microaerophilic streptococci, Haemophilus influenzae and Moraxella catarrhalis. Seventy-one anaerobes were isolated (1.5 per specimen), Peptostreptococcus spp., Prevotella spp., Porphyromonas asaccharolytica and Fusobacterium spp. being predominant. These findings illustrate for the first time the presence of polymicrobial aerobic-anaerobic flora in patients with chronic maxillary sinusitis who had nasal polyposis. PMID- 15888470 TI - Frequency of four different mutacin genes in Streptococcus mutans genotypes isolated from caries-free and caries-active individuals. AB - The ability of Streptococcus mutans to produce mutacins, combined with the production of other virulence factors such as lactic acid, may contribute to the pathogenesis of this bacterium. In the present study, the detection of genes encoding mutacin types I/III, II and IV was performed by PCR with specific primers to each type in a total of 63 S. mutans genotypes isolated from caries active and caries-free individuals. In the caries-free group, PCR screening for mutacin IV revealed that 31.8% of strains were positive for this mutacin. PCR for the other three mutacins tested (I/III and II) did not yield amplicons in any S. mutans strains in this group. The PCR with primers of mutacin IV showed 68.3% positive genotypes in the caries-active group, on the other hand, the amplicons of mutacins I/III revealed 41.5% positive strains that carried these genes. The chi square test showed significant differences in the number of positive strains to mutacin IV when comparing the caries-free and caries-active genotypes of S. mutans (P = 0.01). All tested S. mutans strains were negative by PCR for mutacin II. The low frequencies of detection of some mutacin genes suggest the existence of high diversity and polymorphism in the production of genetic determinants of mutacin-like substances. In addition, the production of a wide spectrum of mutacins can play an important biological role in colonization by S. mutans strains, mainly in the niche of high-complexity microbial communities. PMID- 15888471 TI - Isolation of Escherichia coli O5 :H-, possessing genes for Shiga toxin 1, intimin beta and enterohaemolysin, from an intestinal biopsy from an adult case of bloody diarrhoea: evidence for two distinct O5 :H- pathotypes. AB - Two typical coliforms from an intestinal biopsy from an adult case of bloody diarrhoea carried genes encoding intimin-beta, stx(1) and ehxA, and produced verocytotoxin 1 and enterohaemolysin in culture. Both were biochemically typical Escherichia coli O5 :H(-), apart from producing urease. Such O5 isolates represent a human pathogenic E. coli lineage. PMID- 15888472 TI - Bilateral orbital cellulitis due to Neisseria gonorrhoeae and Staphylococcus aureus: a previously unreported case. AB - Cellulitis of the orbit is a common cause of proptosis in children, and also frequently arises in the elderly and the immunocompromised. The condition is characterized by infection and swelling of the soft tissues lining the eye socket, pushing the eye ball outwards and causing severe pain, redness, discharge of pus and some degree of blurred vision. There is a small risk of infection spreading to the meninges of the brain and causing meningitis. This paper reports the case of an adult in whom polymicrobial bilateral orbital cellulitis had developed due to Staphylococcus aureus and Neisseria gonorrhoeae infection. N. gonorrhoeae infections are acquired by sexual contact. Although the infection may disseminate to a variety of tissues, it usually affects the mucous membranes of the urethra in males and the endocervix and urethra in females. To the authors' knowledge this is the first report of polymicrobial bilateral orbital cellulitis due to S. aureus and N. gonorrhoeae in medical literature. PMID- 15888473 TI - Siderophores--potential candidate in the therapy of neonatal meningitis. PMID- 15888474 TI - Factors associated with mammographic decisions of Chinese-Australian women. AB - BreastScreen (a free breast cancer screening service) has been implemented in Australia since 1991. Surveys conducted overseas consistently report that women of Chinese ancestry have low participation rates in breast cancer screening. Although Chinese women's use of breast cancer screening services has been investigated abroad, to date there are few studies of mammographic screening behavior among Chinese-Australian women. The purpose of this study is to explore and investigate the factors associated with mammographic decisions of Chinese Australian women. Using a qualitative approach, in-depth interviews were conducted with 20 Chinese-Australian women. These were augmented by additional data from ethnographic observations. The findings show two facilitators: organizational factors (an invitation letter from BreastScreen and seniors' clubs arrangements) and the influence of 'significant others'. Barriers identified were fear perceptions of mammography, modesty and fear of stigmatization. This study provides a useful framework for designing and implementing mammographic screening services for Chinese-Australian women that may improve their participation rates. PMID- 15888475 TI - Gene expression in Huntington's disease skeletal muscle: a potential biomarker. AB - Huntington's disease (HD) is an incurable and fatal neurodegenerative disorder. Improvements in the objective measurement of HD will lead to more efficient clinical trials and earlier therapeutic intervention. We hypothesized that abnormalities seen in the R6/2 mouse, a greatly accelerated HD model, might highlight subtle phenotypes in other mouse models and human HD. In this paper, we identify common gene expression changes in skeletal muscle from R6/2 mice, Hdh(CAG(150)) homozygous knock-in mice and HD patients. This HD-triggered gene expression phenotype is consistent with the beginnings of a transition from fast twitch to slow-twitch muscle fiber types. Metabolic adaptations similar to those induced by diabetes or fasting are also present but neither metabolic disorder can explain the full phenotype of HD muscle. The HD-induced gene expression changes reflect disease progression. This raises the possibility that muscle gene expression may be used as an objective biomarker to complement clinical HD-rating systems. Furthermore, an understanding of the molecular basis of muscle dysfunction in HD should provide insight into mechanisms involved in neuronal abnormalities and neurodegeneration. PMID- 15888476 TI - Reduced proportion of Purkinje cells expressing paternally derived mutant Mecp2308 allele in female mouse cerebellum is not due to a skewed primary pattern of X-chromosome inactivation. AB - Rett syndrome (RTT) is an X-linked disorder caused by mutations in the methyl CpG binding protein 2 (MECP2) gene. The pattern of X-chromosome inactivation (XCI) is thought to play a role in phenotypic severity. In the present study, patterns of XCI were assessed by lacZ staining of embryos and adult brains of mice heterozygous for a X-linked Hmgcr-nls-lacZ transgene on a mutant mouse model of RTT. We found that there was no difference between the lacZ staining patterns in the brain of wild-type and heterozygous mutant embryos at embryonic day 9.5 (E9.5) suggesting that Mecp2 has no effect on the primary pattern of XCI. At 20 weeks of age, there was no significant difference between XCI patterns in the Purkinje cells in the cerebellum of heterozygous mutant and wild-type mice when the mutant allele was inherited from the mother. However, when the mutant allele was paternally inherited, a significant difference was detected. Thus, parental origin of the mutation may have a bearing on phenotype through XCI patterns. An estimation of the Purkinje cell precursor number based on XCI mosaicism revealed that, when the mutation was paternally inherited, the precursor number was less than that in the wild-type mice. Therefore, it is likely that the number of precursor cells allocated to the Purkinje cell lineage is affected by a paternally inherited mutation in Mecp2. We also observed that the pattern of XCI in cultured fibroblasts was significantly correlated with patterns in the Purkinje cells in mutant animals but not in wild-type mice. PMID- 15888477 TI - A mouse model of tuberous sclerosis 1 showing background specific early post natal mortality and metastatic renal cell carcinoma. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder caused by mutations in either the TSC1 or the TSC2 genes and characterized by the development of benign hamartomatous growths in multiple organ systems. We have inactivated Tsc1 in the mouse germ line by gene targeting in ES cells and confirmed that the mutant allele (Tsc1-) has a recessive embryonic lethal phenotype. We found that a significant number (approximately 27%) of heterozygous (Tsc1+/-) mice on the C57BL/6 background died before weaning (P = 0.014) and show that these mice die in the post-natal period (P = 0.033), normally at 1-2 days, from unknown causes. Forty-four percent (7/16) of Tsc1+/- mice on a C3H background developed macroscopically visible renal lesions as early as 3-6 months, increasing to 95% (37/39) by 15-18 months. Renal lesions progressed from cysts through cystadenomas to solid carcinomas. Eighty percent (16/20) of Tsc1+/- mice on a Balb/c background exhibited solid renal cell carcinomas (RCC) by 15-18 months and in 41%, RCCs were > or = 5 mm, resulting in grossly deformed kidneys. Some RCCs had a sarcomatoid morphology of spindle cells in whorled patterns and metastasized to the lungs. We detected loss of the wild-type Tsc1 allele and elevated levels of p-mTOR and p-S6 in lesions from Tsc1+/- mice. This new murine model of hamartin deficiency exhibits a more severe phenotype than existing models. PMID- 15888478 TI - Treatment with trkC agonist antibodies delays disease progression in neuromuscular degeneration (nmd) mice. AB - Spinal muscular atrophy with respiratory distress type 1 (SMARD1) is a fatal autosomal recessive disorder seen in infants. It is characterized by lower motor neuron degeneration, progressive muscle paralysis and respiratory failure, for which no effective treatment exists. The phenotype of neuromuscular degeneration (nmd) mice closely resembles the human SMARD1. The identification of the mutated mouse gene in nmd mice, Ighmbp2, led to the discovery of mutations of the homologous gene in humans with SMARD1. We have studied the nmd mouse model with in vivo electrophysiological techniques and evaluated the efficacy of Mab2256, a monoclonal antibody with agonist effect on the tyrosine kinase receptor C, trkC, on disease progression in nmd mice. Treatment with Mab2256 resulted in a significant but transient improvement of muscle strength in nmd mice, as well as normalization of the neuromuscular depression during high-frequency nerve stimulation. These results suggest the potential of using monoclonal agonist antibodies for neurotrophin receptors in lower motor neuron diseases such as SMARD1. PMID- 15888479 TI - Distinct pathogenetic mechanisms for PHOX2B associated polyalanine expansions and frameshift mutations in congenital central hypoventilation syndrome. AB - Congenital central hypoventilation syndrome (CCHS) is a rare neurocristopathy characterized by absence of adequate autonomic control of respiration with decreased sensitivity to hypoxia and hypercapnia. Frameshift mutations and polyalanine triplet expansions in the coding region of PHOX2B have been identified in the vast majority of CCHS patients and a correlation between length of the expanded region and severity of CCHS has been reported. In this work, we have undertaken in vitro analyses aimed at identifying the pathogenetic mechanisms which underlie the effects of PHOX2B mutations in CCHS. According to the known role of this gene, a transcription factor expressed during autonomic nervous system development, we have tested the transcriptional activity of WT and mutant PHOX2B expression constructs on the regulatory regions of two target genes, DbetaH and PHOX2A. We observed that the two sets of mutations play different roles in the transcriptional regulation of these genes, showing a correlation between the length of polyalanine expansions and the severity of reduced transcriptional activity. In particular, although reduced transactivation due to polyalanine expansions may be caused by retention of the mutated protein in the cytoplasm or in the nuclear aggregates, frameshift mutations did not impair the PHOX2B nuclear income, suggesting a different mechanism through which they would exert the observed effects on target promoters. Moreover, the frameshift due to deletion of a cytosine residue seems to cause sequestration of the corresponding mutant PHOX2B in the nucleolar compartment. PMID- 15888480 TI - Haplotypes of the WNK1 gene associate with blood pressure variation in a severely hypertensive population from the British Genetics of Hypertension study. AB - Mutations in the WNK1 gene cause Gordon's syndrome, a rare Mendelian form of hypertension. We assessed whether common WNK1 variants might also contribute to essential hypertension (EH), a multifactorial disorder affecting > 25% of the adult population worldwide. A panel of 19 single nucleotide polymorphisms (SNPs) spanning the gene was selected from public databases and was genotyped in 100 white European families to determine the pattern of linkage disequilibrium, haplotype structure and tagging SNPs for the WNK1 locus. Eight tagging SNPs were identified with 90% power to predict common WNK1 haplotypes and SNPs. Family based association tests were used to test for association with EH and severity of hypertension in 712 severely hypertensive families from the MRC British Genetics of Hypertension study resource. No association was found between WNK1 polymorphisms or haplotypes with hypertension; however, one SNP rs1468326, located 3 kb from the WNK1 promoter, was found to be nominally associated with severity of hypertension, with both systolic blood pressure (BP) (Z = +2.24, P = 0.025) and diastolic BP (Z = +1.99, P = 0.046). We also found nominal support for association of one common WNK1 haplotype with increased systolic BP (Z = +1.91, P = 0.053). This is the first study to perform haplotype association analysis of the WNK1 gene with EH. This finding of association between a SNP near the promoter region and the severity of hypertension suggests that increased expression of WNK1 might contribute to BP variability and susceptibility to EH similar to the mechanism of hypertension observed in Gordon's syndrome. PMID- 15888481 TI - Deletion of VCX-A due to NAHR plays a major role in the occurrence of mental retardation in patients with X-linked ichthyosis. AB - X-linked ichthyosis (XLI) is often associated with a recurrent microdeletion at Xp22.31 due to non-allelic homologous recombination between the CRI-S232 low-copy repeat regions flanking the STS gene. The clinical features of these patients may include mental retardation (MR) and the VCX-A gene has been proposed as the candidate MR gene. Analysis of DNA from four XLI patients with MR by array comparative genomic hybridization (array-CGH) on a 150 kb resolution X chromosome specific array revealed a 1.5 Mb interstitial microdeletion with breakpoints in the CRI-S232 repeat sequences, each of which harbors a VCX gene. We demonstrate that the recombination sites in all four cases are situated in the 1 kb repeat unit 2 region present at the 3' ends of the VCX-A and VCX-B genes thereby deleting VCX-A and VCX-B1 but not VCX-B and VCX-C. Array-CGH with DNA of an XLI patient with MR and an inherited t(X;Y)(p22.31;q11.2) showed an Xpter deletion of 8.0 Mb resulting in the deletion of all four VCX genes and duplication of both VCY homologs. These data confirm the role of VCX-A in the occurrence of MR in XLI patients. Moreover, we propose a VCX/Y teamwork-dependent mechanism for the incidence of mental impairment in XLI patients. PMID- 15888482 TI - Activation of the ALT pathway for telomere maintenance can affect other sequences in the human genome. AB - Immortal human cells maintain telomere length by the expression of telomerase or through the alternative lengthening of telomeres (ALT). The ALT mechanism involves a recombination-like process that allows the rapid elongation of shortened telomeres. However, it is not known whether activation of the ALT pathway affects other sequences in the genome. To address this we have investigated, in ALT-expressing cell lines and tumours, the stability of tandem repeat sequences known to mutate via homologous recombination in the human germline. We have shown extraordinary somatic instability in the human minisatellite MS32 (D1S8) in ALT-expressing (ALT+) but not in normal or telomerase-expressing cell lines. The MS32 mutation frequency varied across 15 ALT+ cell lines and was on average 55-fold greater than in ALT- cell lines. The MS32 minisatellite was also highly unstable in three of eight ALT+ soft tissue sarcomas, indicating that somatic destabilization occurs in vivo. The MS32 mutation rates estimated for two ALT+ cell lines were similar to that seen in the germline. However, the internal structures of ALT and germline mutant alleles are very different, indicating differences in the underlying mutation mechanisms. Five other hypervariable minisatellites did not show elevated instability in ALT expressing cell lines, indicating that minisatellite destabilization is not universal. The elevation of MS32 instability upon activation of the ALT pathway and telomere length maintenance suggests there is overlap between the underlying processes that may be tractable through analysis of the D1S8 locus. PMID- 15888483 TI - Mitochondrial DNA polymerase gamma is essential for mammalian embryogenesis. AB - Mitochondrial DNA (mtDNA) polymerase gamma (Polg) is a heterodimeric enzyme containing a Pol I-like catalytic core (PolgA) and an accessory subunit. Mutations in POLGA, affecting the stability of mtDNA, have been identified in several human pathologies such as progressive external ophthalmoplegia and Alpers' syndrome. Extensive literature shows mitochondrial toxicity effects nucleoside analogue reverse transcriptase inhibitors used in the treatment of HIV and chronic hepatitis B as a consequence of an inhibitory effect on Polg. We have previously shown that mice with an error-prone version of PolgA accumulate higher levels of somatic mtDNA mutations resulting in a premature aging phenotype. In the present paper, we demonstrate PolgA deficiency in mouse embryos causes an early developmental arrest between embryonic days 7.5 and 8.5 associated with severe mtDNA depletion. Heterozygous knockout mice have half the wild-type levels of PolgA transcripts and a slight reduction in mtDNA levels but develop normally. Surprisingly, amounts of PolgA transcripts in heterozygous knockout mice are increased in response to artificially elevated mtDNA copy number, revealing a possible regulatory link between mtDNA maintenance and PolgA expression. Our results show that Polg indeed is the only DNA polymerase capable of maintaining mtDNA in mammalian mitochondria. In addition, presence of Polg is absolutely essential for the organogenesis during mammalian embryonic development. PMID- 15888484 TI - Disruption of Abcc6 in the mouse: novel insight in the pathogenesis of pseudoxanthoma elasticum. AB - Pseudoxanthoma elasticum (PXE) is a heritable disorder of connective tissue, affecting mainly skin, eye and the cardiovascular system. PXE is characterized by dystrophic mineralization of elastic fibres. The condition is caused by loss of function mutations in ABCC6. We generated Abcc6 deficient mice (Abcc6-/-) by conventional gene targeting. As shown by light and electron microscopy Abcc6-/- mice spontaneously developed calcification of elastic fibres in blood vessel walls and in Bruch's membrane in the eye. No clear abnormalities were seen in the dermal extracellular matrix. Calcification of blood vessels was most prominent in small arteries in the cortex of the kidney, but in old mice, it occurred also in other organs and in the aorta and vena cava. Newly developed monoclonal antibodies against mouse Abcc6 localized the protein to the basolateral membranes of hepatocytes and the basal membrane in renal proximal tubules, but failed to show the protein at the pathogenic sites. Abcc6-/- mice developed a 25% reduction in plasma HDL cholesterol and an increase in plasma creatinine levels, which may be due to impaired kidney function. No changes in serum mineral balance were found. We conclude that the phenotype of the Abcc6-/- mouse shares calcification of elastic fibres with human PXE pathology, which makes this model a useful tool to further investigate the aetiology of PXE. Our data support the hypothesis that PXE is in fact a systemic disease. PMID- 15888485 TI - Genomic architecture of human 17q21 linked to frontotemporal dementia uncovers a highly homologous family of low-copy repeats in the tau region. AB - Familial frontotemporal dementia (FTD), characterized by tau-negative, ubiquitin positive inclusions at autopsy, is linked to a chromosomal region at 17q21 (FTDU 17), encompassing the gene encoding the microtubule associated protein tau, MAPT. Mutations in MAPT were previously identified in familial FTD with parkinsonism (FTDP-17); however, in FTDU-17 patients, no pathogenic mutations were found in exonic regions consistent with the lack of tauopathy in FTDU-17 brains. Here, we excluded mutations in MAPT by genomic sequencing of 138.5 kb in FTDU-17 patients. Next, to facilitate the identification of the actual underlying genetic defect, we assembled the 6.5 Mb FTDU-17 sequence. Annotation demonstrated that MAPT is surrounded by three highly homologous low-copy repeats (LCRs) in a region of 1.7 Mb. Using evolutionary studies, short tandem repeat-based linkage disequilibrium (LD) and macro-restriction mapping, we demonstrated that these LCRs are at the basis of a series of rearrangements in the MAPT genomic region. One is an inversion that occurred 3 million years ago and resulted in a common polymorphism in humans to date. This inversion plus flanking LCRs spanned approximately 1.3 Mb and was shown to underlie the extended LD and haplotypes H1 and H2 across MAPT. However, in the FTDU-17 families, we ascertained segregation analysis precluding a relationship between the FTDU-17 and the H1/H2 inversion. The presence of multiple homologous LCRs in the region predicts that other potentially more complex genomic rearrangements might be underlying FTDU-17. PMID- 15888486 TI - Cancer chemoprevention by the antioxidant tempol acts partially via the p53 tumor suppressor. AB - We previously demonstrated that the nitroxide antioxidant tempol (4-hydroxy 2,2,6,6-tetramethylpiperidine-N-oxyl) increased latency to tumorigenesis and doubled (100%) the lifespan of Atm-deficient mice, a mouse model of ataxia telangiectasia, which displays accelerated oxidative damage and stress. Tempol treatment of cancer-prone p53-deficient mice resulted in a small but significant (25%) increase in lifespan by prolonging latency to tumorigenesis, demonstrating that existing oxidative stress and damage are not necessary for the chemopreventative effects of tempol. However, the relatively small effect on latency in p53-deficient mice and the finding that tempol-mediated resistance to oxidative insult was p53-dependent suggested a more direct role of p53 in the chemopreventative effects of tempol. Surprisingly, tempol treatment specifically increased serine 18 phosphorylation of p53 (but not gamma-H2AX) and p21 expression in primary thymocytes in vitro in a p53-dependent fashion. Inhibition of phosphoinositide 3-kinase (PI3K) family members suggested that SMG-1 was responsible for the tempol-mediated enhancement of p53 serine 18 phosphorylation. These data suggest that the chemopreventative effect of tempol is not solely due to the reduction of oxidative stress and damage but may also be related to redox mediated signaling functions that include p53 pathway activation. PMID- 15888487 TI - A novel functional VKORC1 promoter polymorphism is associated with inter individual and inter-ethnic differences in warfarin sensitivity. AB - Warfarin, a commonly prescribed anticoagulant, exhibited large inter-individual and inter-ethnic differences in the dose required for its anticoagulation effect. Asian populations, including Chinese, require a much lower maintenance dose than Caucasians, for which the mechanisms still remain unknown. We determined DNA sequence variants in CYP2C9 and VKORC1 in 16 Chinese patients having warfarin sensitivity (< or = 1.5 mg/day, n = 11) or resistance (> or = 6.0 mg/day, n = 5), 104 randomly selected Chinese patients receiving warfarin, 95 normal Chinese controls and 92 normal Caucasians. We identified three CYP2C9 variants, CYP2C9*3, T299A and P382L, in four warfarin-sensitive patients. A novel VKORC1 promoter polymorphism (-1639 G > A) presented in the homozygous form (genotype AA) was found in all warfarin-sensitive patients. The resistant patients were either AG or GG. Among the 104 randomly selected Chinese patients receiving warfarin, AA genotype also had lower dose than the AG/GG genotype (P < 0.0001). Frequencies of AA, AG and GG genotypes were comparable in Chinese patients receiving warfarin (79.7, 17.6 and 2.7%) and normal Chinese controls (82, 18 and 0%), but differed significantly from Caucasians (14, 47 and 39%) (P < 0.0001). The promoter polymorphism abolished the E-box consensus sequences and dual luciferase assay revealed that VOKRC1 promoter with the G allele had a 44% increase of activity when compared with the A allele. The differences in allele frequencies of A/G allele and its levels of VKORC1 promoter activity may underscore the inter individual differences in warfarin dosage as well as inter-ethnic differences between Chinese and Caucasians. PMID- 15888488 TI - Zebrafish as a model for caveolin-associated muscle disease; caveolin-3 is required for myofibril organization and muscle cell patterning. AB - Caveolae are an abundant feature of many animal cells. However, the exact function of caveolae remains unclear. We have used the zebrafish, Danio rerio, as a system to understand caveolae function focusing on the muscle-specific caveolar protein, caveolin-3 (Cav3). We have identified caveolin-1 (alpha and beta), caveolin-2 and Cav3 in the zebrafish. Zebrafish Cav3 has 72% identity to human CAV3, and the amino acids altered in human muscle diseases are conserved in the zebrafish protein. During embryonic development, cav3 expression is apparent by early segmentation stages in the first differentiating muscle precursors, the adaxial cells and slightly later in the notochord. cav3 expression appears in the somites during mid-segmentation stages and then later in the pectoral fins and facial muscles. Cav3 and caveolae are located along the entire sarcolemma of late stage embryonic muscle fibers, whereas beta-dystroglycan is restricted to the muscle fiber ends. Down-regulation of Cav3 expression causes gross muscle abnormalities and uncoordinated movement. Ultrastructural analysis of isolated muscle fibers reveals defects in myoblast fusion and disorganized myofibril and membrane systems. Expression of the zebrafish equivalent to a human muscular dystrophy mutant, CAV3P104L, causes severe disruption of muscle differentiation. In addition, knockdown of Cav3 resulted in a dramatic up-regulation of eng1a expression resulting in an increase in the number of muscle pioneer-like cells adjacent to the notochord. These studies provide new insights into the role of Cav3 in muscle development and demonstrate its requirement for correct intracellular organization and myoblast fusion. PMID- 15888490 TI - Complicating the complexity of p53. AB - Recent studies have suggested that the straightforward role of p53 as a transcription factor that functions by inducing apoptotic target genes to eliminate developing tumor cells is only part of a much more complicated story. There is now a firm body of evidence supporting a transcriptionally independent activity of p53 as a functional, if not structural, homologue of the BH3-only proteins. Although this information adds another nuance to the mechanism by which p53 can induce apoptosis, further studies indicate that the apoptotic function of p53 represents only a part of its tumor suppressive activity. Although complicating our understanding of p53, these new insights may also provide some exciting new targets for the design of therapeutics that can reactivate p53 in cancers. PMID- 15888489 TI - Cell type-specific gene expression of midbrain dopaminergic neurons reveals molecules involved in their vulnerability and protection. AB - Molecular differences between dopamine (DA) neurons may explain why the mesostriatal DA neurons in the A9 region preferentially degenerate in Parkinson's disease (PD) and toxic models, whereas the adjacent A10 region mesolimbic and mesocortical DA neurons are relatively spared. To characterize innate physiological differences between A9 and A10 DA neurons, we determined gene expression profiles in these neurons in the adult mouse by laser capture microdissection, microarray analysis and real-time PCR. We found 42 genes relatively elevated in A9 DA neurons, whereas 61 genes were elevated in A10 DA neurons [> 2-fold; false discovery rate (FDR) < 1%]. Genes of interest for further functional analysis were selected by criteria of (i) fold differences in gene expression, (ii) real-time PCR validation and (iii) potential roles in neurotoxic or protective biochemical pathways. Three A9-elevated molecules [G protein coupled inwardly rectifying K channel 2 (GIRK2), adenine nucleotide translocator 2 (ANT-2) and the growth factor IGF-1] and three A10-elevated peptides (GRP, CGRP and PACAP) were further examined in both alpha-synuclein overexpressing PC12 (PC12-alphaSyn) cells and rat primary ventral mesencephalic (VM) cultures exposed to MPP+ neurotoxicity. GIRK2-positive DA neurons were more vulnerable to MPP+ toxicity and overexpression of GIRK2 increased the vulnerability of PC12-alphaSyn cells to the toxin. Blocking of ANT decreased vulnerability to MPP+ in both cell culture systems. Exposing cells to IGF-1, GRP and PACAP decreased vulnerability of both cell types to MPP+, whereas CGRP protected PC12-alphaSyn cells but not primary VM DA neurons. These results indicate that certain differentially expressed molecules in A9 and A10 DA neurons may play key roles in their relative vulnerability to toxins and PD. PMID- 15888491 TI - Regulation of AKT1 expression by beta-catenin/Tcf/Lef signaling in colorectal cancer cells. AB - The serine/threonine kinase AKT plays a critical role in controlling the balance between cell survival and apoptosis. Several reports implicated AKT in the molecular pathogenesis of different human malignancies and overexpression of AKT was recently demonstrated to be an early event in colorectal carcinogenesis. We report here the identification of nine putative Tcf/Lef-binding elements (TBEs) upstream to the ATG initiation site of the AKT1 gene. Four of these TBEs are located upstream of the transcriptional start, whereas five TBEs are situated in Exon 1 of the AKT1 gene. Accordingly, we hypothesized that AKT1 expression might be regulated by Wnt/beta-catenin signaling. To elucidate the regulation of AKT expression in colon cancer cells, we generated reporter constructs containing the luciferase gene under the control of different regions derived from the AKT1 promoter/enhancer. Transient expression of the constructs in colorectal cancer (CRC) cell lines resulted in significant activation of the reporter gene. Luciferase was stimulated 20- to 50-fold in SW480, SW948 and HCT116 CRC cells. In contrast, the AKT1 promoter/enhancer constructs showed only a weak response in 293 embryonic kidney cells. Coexpression of a constitutively active beta-catenin mutant in colon cancer cells further enhanced reporter gene activation from the AKT1 promoter/enhancer, whereas it was downregulated by introduction of either wild-type APC or dnTcf-4. In addition, immunohistochemical staining of tumor sections derived from CRC patients showed elevated expression levels of AKT1, correlating with enhanced cytoplasmic/nuclear expression of beta-catenin. In summary our data suggest that beta-catenin/Tcf contributes to the transcriptional regulation of the AKT1 gene. PMID- 15888492 TI - Opposing effects of prepubertal low- and high-fat n-3 polyunsaturated fatty acid diets on rat mammary tumorigenesis. AB - To determine whether dietary fat intake during childhood affects the later risk of developing breast cancer, we fed prepubertal rats between post-natal days 5 and 25 a low (16% energy) or high-fat (39% energy) diet composed mainly of n-6 or n-3 polyunsaturated fatty acids (PUFAs) originating either from corn oil or menhaden oil, respectively, in the ratios of 16-17:1 (n-6 PUFA diets) or 2-3:1 (n 3 PUFA diets). We also examined whether changes in risk are associated with perturbations in biological processes previously linked to fatty acid intake and breast cancer. Mammary tumorigenesis was induced by treating 50-day-old rats with the carcinogen 7,12-dimethylbenz[a]anthracene. When compared with the reference low-fat n-6 PUFA diet, prepubertal exposure to the low-fat n-3 PUFA diet decreased, whereas a high-fat n-3 PUFA diet increased mammary tumor incidence; the high-fat n-6 PUFA diet had no effect. Both the low and high-fat n-3 PUFA diets induced mammary epithelial differentiation by reducing the number of terminal end buds (TEBs) and increasing the presence of lobulo-alveolar structures. They also increased lipid peroxidation and reduced cyclooxygenase-2 activity. Prepubertal exposure to the low-fat n-3 PUFA diet increased apoptosis, determined using TUNEL assay, and reduced cell proliferation, determined using PCNA staining. In marked contrast, prepubertal exposure to the high-fat n-3 PUFA diet induced cell proliferation and inhibited apoptosis in the TEBs and lobular structures. The latter is consistent with the finding that pAkt, a survival factor that inhibits apoptosis, was elevated in their mammary glands. In summary, although prepubertal exposure to a low-fat n-3 PUFA diet reduced later mammary tumorigenesis in rats, high levels of this fatty acid can have adverse effects on the prepubertal mammary gland and increase subsequent breast cancer risk. PMID- 15888493 TI - Inhibition of chemically-induced neoplastic transformation by a novel tetrasodium diphosphate astaxanthin derivative. AB - Carotenoids have been implicated in numerous epidemiological studies as being protective against cancer at many sites, and their chemopreventive properties have been confirmed in laboratory studies. Astaxanthin (AST), primarily a carotenoid of marine origin, responsible for the pink coloration of salmon, shrimp and lobster, has received relatively little attention. As with other carotenoids, its highly lipophilic properties complicate delivery to model systems. To overcome this issue we have synthesized a novel tetrasodium diphosphate astaxanthin (pAST) derivative with aqueous dispersibility of 25.21 mg/ml. pAST was delivered to C3H/10T1/2 cells in an aqueous/ethanol solution and compared with non-esterified AST dissolved in tetrahydrofuran. We show pAST to (i) upregulate connexin 43 (Cx43) protein expression; (ii) increase the formation of Cx43 immunoreactive plaques; (iii) upregulate gap junctional intercellular communication (GJIC); and (iv) cause 100% inhibition of methylcholanthrene induced neoplastic transformation at 10(-6) M. In all these assays, pAST was superior to non-esterified AST itself; in fact, pAST exceeded the potency of all other previously tested carotenoids in this model system. Cleavage of pAST to non esterified (free) AST and uptake into cells was also verified by HPLC; however, levels of free AST were approximately 100-fold lower than in cells treated with AST itself, suggesting that pAST possesses intrinsic activity. The dual properties of water dispersibility (enabling parenteral administration in vivo) and increased potency should prove extremely useful in the future development of cancer chemopreventive agents. PMID- 15888494 TI - Quantification of DNA and hemoglobin adducts of 3,4-epoxy-1,2-butanediol in rodents exposed to 3-butene-1,2-diol. AB - 1,3-Butadiene (BD) is a confirmed rodent carcinogen and a suspect human carcinogen that forms mutagenic epoxide metabolites during biotransformation. Species differences in the roles of individual DNA reactive intermediates in BD mutagenicity and carcinogenicity are not completely understood. Evidence suggests that 1,2:3,4-diepoxybutane (DEB) is responsible for the mutagenic effect induced by exposures to low concentrations of BD in mice and that metabolites of 3-butene 1,2-diol (BD-diol) are involved in the mutagenicity at high exposures in both mice and rats. Two reactive metabolites, 3,4-epoxy-1,2-butanediol (EB-diol) and hydroxymethylvinyl ketone (HMVK), are formed during the biotransformation of BD diol and could potentially be involved in BD-diol associated mutagenicity. To examine the role of EB-diol in BD-diol mutagenicity we have evaluated the dosimetry of N7-(2,3,4-trihydroxybutyl)guanine (THB-Gua) and N-(2,3,4 trihydroxybutyl)valine (THB-Val) in female B6C3F1 mice and female F344 rats exposed by inhalation to 0, 6, 18 and 36 p.p.m. BD-diol for 4 weeks (6 h/day x 5 days/week). Results showed higher levels of both THB-Gua and THB-Val in mice than in rats. An evaluation of THB-Gua adducts showed virtually no differences between liver and lung for either species, suggesting that EB-diol is stable and is freely circulated. The data also indicated that THB adduct formation began to plateau around 18 p.p.m. in both species. Most importantly, the shape of the dose response curve for THB adduct formation mimicked the one observed for hypoxanthine-guanine phosphoribosyltransferase (Hprt) mutation frequency. This showed that THB adducts, which are not thought to be responsible for causing the mutations, are good quantitative indicators of mutagenicity in rodents exposed to BD-diol. Although the potential contribution of HMVK still needs to be evaluated, the data suggest that EB-diol is responsible, at least in part, for BD-diol associated mutagenicity in rodents. PMID- 15888495 TI - Effects of dietary folate on the development and progression of mammary tumors in rats. AB - Epidemiologic studies have suggested that dietary intake and blood levels of folate may be inversely related to the risk of breast cancer. However, epidemiologic evidence has not been consistent nor has it provided unequivocal support for this purported inverse relationship. Recent evidence has also raised a concern that folate supplementation may promote carcinogenesis if provided after neoplastic foci are established in the target organ. This study investigated the effect of dietary folate deficiency and supplementation on the development and progression of mammary tumors in the N-methyl-N-nitrosourea (MNU) rat model. Weanling, female Sprague-Dawley rats were fed diets containing 0, 2 (control) or 8 mg folic acid/kg diet during the initiation or the promotion phase of MNU-induced mammary tumorigenesis. At necropsy, all macroscopic mammary tumors were identified and histologically confirmed. Dietary folate deficiency and supplementation provided during the initiation phase did not significantly modulate the development of mammary tumors. In contrast, dietary folate deficiency provided during the promotion phase significantly inhibited the rate of appearance, incidence, mean volume and weight of adenocarcinomas compared with the control and supplemental diets. Folate supplementation provided during the promotion phase did not significantly modulate mammary tumorigenesis compared with the control group. These data indicate that moderate folate deficiency inhibits, whereas dietary folate supplementation at four times the basal dietary requirement does not promote, the progression of MNU-induced mammary neoplastic foci in this rat model. However, the limitations associated with the route and dose of MNU administration preclude a definitive conclusion concerning the effect of folate status on the initiation of MNU-induced mammary tumorigenesis. PMID- 15888496 TI - Clinical efficacy and survival with first-line inhaled iloprost therapy in patients with idiopathic pulmonary arterial hypertension. AB - AIMS: To describe the long-term clinical efficacy of inhaled iloprost as first line vasodilator mono-therapy in patients with idiopathic pulmonary arterial hypertension (IPAH). METHODS AND RESULTS: Seventy-six IPAH patients were prospectively identified and treated with inhaled iloprost. Clinical, haemodynamic, and exercise parameters were obtained at baseline, after 3 and 12 months of therapy and yearly thereafter. Four endpoints were prospectively defined as follows: (i) death, (ii) transplantation, (iii) switch to intravenous (i.v.) therapy, or (iv) addition of or switch to other active oral therapy. During follow-up (535+/-61 days), 11 patients died, six were transplanted, 25 were switched to i.v. prostanoids, 16 received additional or other oral therapy, and 12 patients discontinued iloprost inhalation for other reasons. Event-free survival at 3, 12, 24, 36, 48, and 60 months was 81, 53, 29, 20, 17 and 13%, respectively. Among haemodynamic and exercise parameters, mixed venous oxygen saturation (P<0.001), right atrial pressure (P<0.001), and peak oxygen uptake (P=0.002) were associated with event-free survival. CONCLUSION: In this study, only a minority of patients could be stabilized with inhaled iloprost mono therapy during a follow-up period of up to 5 years. In the presence of multiple treatment options, chronic iloprost inhalation as mono-therapy appears to have a limited role. PMID- 15888497 TI - Non-cardiac QTc-prolonging drugs and the risk of sudden cardiac death. AB - AIMS: To assess the association between the use of non-cardiac QTc-prolonging drugs and the risk of sudden cardiac death. METHODS AND RESULTS: A population based case-control study was performed in the Integrated Primary Care Information (IPCI) project, a longitudinal observational database with complete medical records from more than 500,000 persons. All deaths between 1 January 1995 and 1 September 2003 were reviewed. Sudden cardiac death was classified based on the time between onset of cardiovascular symptoms and death. For each case, up to 10 random controls were matched for age, gender, date of sudden death, and general practice. The exposure of interest was the use of non-cardiac QTc-prolonging drugs. Exposure at the index date was categorized into three mutually exclusive groups of current use, past use, and non-use. The study population comprised 775 cases of sudden cardiac death and 6297 matched controls. Current use of any non cardiac QTc-prolonging drug was associated with a significantly increased risk of sudden cardiac death (adjusted OR: 2.7; 95% CI: 1.6-4.7). The risk of death was highest in women and in recent starters. CONCLUSION: The use of non-cardiac QTc prolonging drugs in a general population is associated with an increased risk of sudden cardiac death. PMID- 15888498 TI - Non-invasive computed tomographic coronary angiography: the end of the beginning. PMID- 15888499 TI - Angiographic, intravascular ultrasound, and fractional flow reserve evaluation of direct stenting vs. conventional stenting using BeStent2 in a multicentre randomized trial. AB - AIMS: Direct stenting (DS) may not be as safe and effective as conventional stenting. The objective was to demonstrate equivalence of post-procedural mean luminal diameter (MLD) by angiography after BeStent2 placement between DS and pre dilatation (PD) strategy. METHODS AND RESULTS: Two hundred and two patients with a single de novo lesion (diameter >/=3.0 mm and length or = 100 microM in the immediate vicinity of the vesicle. This [Ca2+] is much higher than some of the recent estimates for the equilibrium dissociation constant of the Ca2+ sensor(s) that control neurotransmitter release, suggesting release should be close to saturation, yet it is well known that release is highly sensitive to changes in Ca2+ influx. We show that due to the brevity of the Ca2+ influx the binding kinetics of the Ca2+ sensor rather than its equilibrium affinity determine receptor occupancy. For physiologically relevant Ca2+ currents and forward Ca2+ binding rates, the effective affinity of the Ca2+ sensor can be several-fold lower than the equilibrium affinity. Using simple models, we show redundant copies of the binding sites increase effective affinity of the Ca2+ sensor for release. Our results predict that different levels of expression of Ca2+ binding sites could account for apparent differences in Ca2+ sensor affinities between synapses. Using Monte Carlo simulations of Ca2+ dynamics with nanometer resolution, we demonstrate that these kinetic constraints combined with vesicles acting as diffusion barriers can prevent saturation of the Ca2+-sensor(s) for neurotransmitter release. We further show the random positioning of the Ca2+-sensor molecules around the vesicle can result in the emergence of two distinct populations of the vesicles with low and high release probability. These considerations allow experimental evidence for the Ca2+ channel-vesicle colocalization to be reconciled with a high equilibrium affinity for the Ca2+ sensor of the release machinery. PMID- 15888527 TI - Duration selectivity of neurons in the inferior colliculus of the big brown bat: tolerance to changes in sound level. AB - At and above the level of the inferior colliculus (IC), some neurons respond maximally to a limited range of sound durations, with little or no excitatory response to durations outside of this range. Such neurons have been termed "duration tuned" or "duration selective." In this study we examined the effects of varying signal amplitude on best duration, width of tuning, and first spike latency of duration tuned neurons in the IC of the big brown bat, Eptesicus fuscus. Response areas as a function of stimulus duration and intensity took a variety of forms, including open (V-shaped), narrow and level tolerant (U shaped), or closed (O-shaped). The majority (82%) of duration tuned neurons had narrow U-shaped or O-shaped duration response areas. Those with narrow U-shaped response areas retained their duration tuning across a broad dynamic range, < or = 50 dB above threshold, whereas those with O-shaped response areas were narrowly tuned to both stimulus duration and amplitude. For about one-half (55%) of the neurons with either a U- or O-shaped response areas, best duration (BD) changed by <1 ms across the range of suprathreshold amplitudes tested. Changes in BD most often took the form of a shift to slightly shorter durations as stimulus level increased. For the majority (65%) of U- and O-shaped neurons, 50% width of duration tuning changed by <2 ms with increasing amplitude. Latency of response at BD remained stable across changes in sound level, suggesting that the relative strengths of excitatory and inhibitory inputs to duration tuned neurons remain in balance over a wide dynamic range of sound pressure levels. PMID- 15888528 TI - Postnatal development differentially affects voltage-activated calcium currents in respiratory rhythmic versus nonrhythmic neurons of the pre-Botzinger complex. AB - The mammalian respiratory network reorganizes during early postnatal life. We characterized the postnatal developmental changes of calcium currents in neurons of the pre-Botzinger complex (pBC), the presumed site for respiratory rhythm generation. The pBC contains not only respiratory rhythmic (R) but also nonrhythmic neurons (nR). Both types of neurons express low- and high-voltage activated (LVA and HVA) calcium currents. This raises the interesting issue: do calcium currents of the two co-localized neuron types have similar developmental profiles? To address this issue, we used the whole cell patch-clamp technique to compare in transverse slices of mice LVA and HVA calcium current amplitudes of the two neuron populations (R and nR) during the first and second postnatal week (P0-P16). The amplitude of HVA currents did not significantly change in R pBC neurons (P0-P16), but it significantly increased in nR pBC-neurons during P8-P16. The dehydropyridine (DHP)-sensitive current amplitudes did not significantly change during the early postnatal development, suggesting that the observed amplitude changes in nR pBC-neurons are caused by (DHP) insensitive calcium currents. The ratio between HVA calcium current amplitudes dramatically changed during early postnatal development: At P0-P3, current amplitudes were significantly larger in R pBC-neurons, whereas at P8-P16, current amplitudes were significantly larger in nR pBC-neurons. Our results suggest that calcium currents in pBC neurons are differentially altered during postnatal development and that R pBC-neurons have fully expressed calcium currents early during postnatal development. This may be critical for stable respiratory rhythm generation in the underlying rhythm generating network. PMID- 15888529 TI - Inhibition of perforant path input to the CA1 region by serotonin and noradrenaline. AB - Bath-applied monoamines-dopamine (DA), serotonin (5-HT), and noradrenaline (NE) strongly suppress the perforant path (PP) input to CA1 hippocampal region with very little effect on the Schaffer collaterals (SC) input. The effect of DA action on PP field excitatory postsynaptic potential (fEPSP) has been characterized in detail, but relatively little is known about the NE and 5-HT effects. Here we show that the maximal inhibition of the PP fEPSP by NE is approximately 55%, whereas 5-HT inhibition is weaker ( approximately 35%). The half-maximal inhibitory concentration of both 5-HT and NE is approximately 1 muM. Neither NE nor 5-HT affected paired-pulse facilitation, suggesting that the effect is not presynaptic. This is in contrast to DA, which does have a presynaptic effect. The NE effect was blocked by alpha2 antagonists, whereas the alpha1 antagonist corynanthine and beta-antagonist propranolol were ineffective. The effect of 5-HT was mimicked by the agonist, 5-carboxamidotryptamine maleate (5-CT), and not affected by adrenergic and dopaminergic antagonists. To determine the 5-HT receptors involved, we tested a number of 5-HT antagonists, but none produced a complete suppression of the 5-HT effect. Of these, only the 5-HT7 and 5-HT2 antagonists produced weak but significant inhibition of 5-HT effect. We conclude that NE inhibits the PP fEPSP through postsynaptic action on alpha2 adrenoceptors and that 5-HT7, 5-HT2, and some other receptor may be involved in 5 HT action in PP. PMID- 15888530 TI - Spatiotemporal patterns of dorsal root-evoked network activity in the neonatal rat spinal cord: optical and intracellular recordings. AB - Spatiotemporal patterns of dorsal root-evoked potentials were studied in transverse slices of the rat spinal cord by monitoring optical signals from a voltage-sensitive dye with multiple-photodiode optic camera. Typically, dorsal root stimulation generated two basic waveforms of voltage images: dual-component images consisting of fast, spike-like signal followed by a slow signal in the dorsal horn, and small, slow signals in the ventral horn. To qualitatively relate the optical signals to membrane potentials, whole cell recordings were combined with measurements of light absorption in the area around the soma. The slow optical signals correlated closely with subthreshold postsynaptic potentials in all regions of the cord. The spike-like component was not associated with postsynaptic action potentials, suggesting that the fast signal was generated by presynaptic action potentials. Firing in a single neuron could not be detected optically, implying that local voltage images originated from synchronously activated neuronal ensembles. Blocking glutamatergic synaptic transmission inhibited excitatory postsynaptic potentials (EPSPs) and significantly reduced the slow optical signals, indicating that they were mediated by glutamatergic synapses. Suppressing glycine-mediated inhibition increased the amplitude of both optical signals and EPSPs, while blocking GABA(A) receptor-mediated synapses, increased the amplitude and time course of EPSPs and prolonged the duration of voltage images in larger areas of the slice. The close correlation between evoked EPSPs and their respective local voltage images shows the advantage of the high temporal resolution optical system in measuring both the spatiotemporal dynamics of segmental network excitation and integrated potentials of neuronal ensembles at identified sites. PMID- 15888531 TI - Trajectory interpretation by supplementary eye field neurons during ocular baseball. AB - Good performance in the sport of baseball shows that humans can determine the trajectory of a moving object and act on it under the constraint of a rule. We report here on neuronal activity in the supplementary eye field (SEF) of monkeys performing an eye movement task inspired by baseball. In "ocular baseball," a pursuit eye movement to a target is executed or withheld based on the target's trajectory. We found that a subset of neurons in the SEF interpreted the trajectory according to the task rule. Other neurons specified at a later time the command to pursue the target with the eyes. The results suggest that the SEF can interpret sensory signals about target motion in the context of a rule to guide voluntary eye movement initiation. PMID- 15888532 TI - AP180 maintains the distribution of synaptic and vesicle proteins in the nerve terminal and indirectly regulates the efficacy of Ca2+-triggered exocytosis. AB - AP180 plays an important role in clathrin-mediated endocytosis of synaptic vesicles (SVs) and has also been implicated in retrieving SV proteins. In Drosophila, deletion of its homologue, Like-AP180 (LAP), has been shown to increase the size of SVs but decrease the number of SVs and transmitter release. However, it remains elusive whether a reduction in the total vesicle pool directly affects transmitter release. Further, it is unknown whether the lap mutation also affects vesicle protein retrieval and synaptic protein localization and, if so, how it might affect exocytosis. Using a combination of electrophysiology, optical imaging, electron microscopy, and immunocytochemistry, we have further characterized the lap mutant and hereby show that LAP plays additional roles in maintaining both normal synaptic transmission and protein distribution at synapses. While increasing the rate of spontaneous vesicle fusion, the lap mutation dramatically reduces impulse-evoked transmitter release at steps downstream of calcium entry and vesicle docking. Notably, lap mutations disrupt calcium coupling to exocytosis and reduce calcium cooperativity. These results suggest a primary defect in calcium sensors on the vesicles or on the release machinery. Consistent with this hypothesis, three vesicle proteins critical for calcium-mediated exocytosis, synaptotagmin I, cysteine-string protein, and neuronal synaptobrevin, are all mislocalized to the extrasynaptic axonal regions along with Dap160, an active zone marker (nc82), and glutamate receptors in the mutant. These results suggest that AP180 is required for either recycling vesicle proteins and/or maintaining the distribution of both vesicle and synaptic proteins in the nerve terminal. PMID- 15888533 TI - Dynamic circuitry for updating spatial representations. II. Physiological evidence for interhemispheric transfer in area LIP of the split-brain macaque. AB - With each eye movement, a new image impinges on the retina, yet we do not notice any shift in visual perception. This perceptual stability indicates that the brain must be able to update visual representations to take our eye movements into account. Neurons in the lateral intraparietal area (LIP) update visual representations when the eyes move. The circuitry that supports these updated representations remains unknown, however. In this experiment, we asked whether the forebrain commissures are necessary for updating in area LIP when stimulus representations must be updated from one visual hemifield to the other. We addressed this question by recording from LIP neurons in split-brain monkeys during two conditions: stimulus traces were updated either across or within hemifields. Our expectation was that across-hemifield updating activity in LIP would be reduced or abolished after transection of the forebrain commissures. Our principal finding is that LIP neurons can update stimulus traces from one hemifield to the other even in the absence of the forebrain commissures. This finding provides the first evidence that representations in parietal cortex can be updated without the use of direct cortico-cortical links. The second main finding is that updating activity in LIP is modified in the split-brain monkey: across-hemifield signals are reduced in magnitude and delayed in onset compared with within-hemifield signals, which indicates that the pathways for across hemifield updating are less effective in the absence of the forebrain commissures. Together these findings reveal a dynamic circuit that contributes to updating spatial representations. PMID- 15888534 TI - Dynamic circuitry for updating spatial representations. I. Behavioral evidence for interhemispheric transfer in the split-brain macaque. AB - Internal representations of the sensory world must be constantly adjusted to take movements into account. In the visual system, spatial updating provides a mechanism for maintaining a coherent map of salient locations as the eyes move. Little is known, however, about the pathways that produce updated spatial representations. In the present study, we asked whether direct cortico-cortical links are required for spatial updating. We addressed this question by investigating whether the forebrain commissures-the direct path between the two cortical hemispheres-are necessary for updating visual representations from one hemifield to the other. We assessed spatial updating in two split-brain monkeys using the double-step task, which involves saccades to two sequentially appearing targets. Accurate performance requires that the representation of the second target be updated to take the first saccade into account. We made two central discoveries regarding the pathways that underlie spatial updating. First, we found that split-brain monkeys exhibited a selective initial impairment on double step sequences that required updating across visual hemifields. Second, and most surprisingly, these impairments were neither universal nor permanent: the monkeys were ultimately able to perform the across-hemifield sequences and, in some cases, this ability emerged rapidly. These findings indicate that direct cortical links provide the main substrate for updating visual representations, but they are not the sole substrate. Rather, a unified and stable representation of visual space is supported by a redundant cortico-subcortical network with a striking capacity for reorganization. PMID- 15888535 TI - Adapting locomotion to different surface compliances: neuromuscular responses and changes in movement dynamics. AB - Knowledge of how the nervous system deals with surfaces with different physical properties such as compliance that challenge balance during locomotion is of importance as we are constantly faced with these situations every day. The purpose of this study was to examine the control of center of mass (COM) and lower limb dynamics and recovery response modulation of muscle activity during locomotion across an unexpected compliant surface and in particular, scaling behavior across different levels of compliance. Eight young adults walked along a walkway and stepped on an unexpected compliant surface in the middle of the travel path. There were three different levels of surface compliance, and participants experienced either no compliant surface or one of the three compliant surfaces during each trial that were presented in a blocked or random fashion. Whole body kinematics were collected along with surface electromyography (EMG) of selected bilateral lower limb and trunk muscles. The recovery response to the first compliant-surface trial demonstrated muscle onset latencies between 97 and 175 ms, and activity was modulated while on the compliant surface. Vertical COM trajectory was not preserved after contact with the compliant surface: peak vertical COM, while on the compliant surface was lower than when on stable ground. Perturbed-limb knee flexion after toe-off increased with increased surface compliance, which enabled toe clearance with the ground to be similar to control trials. The results suggest that stepping off of a compliant surface is actively modulated by the CNS and is geared toward maintaining dynamic stability. PMID- 15888536 TI - Glucose metabolism in early onset versus late onset Alzheimer's disease: an SPM analysis of 120 patients. AB - The aims of this cross-sectional study were (i) to compare the overall glucose metabolism between early onset and late onset Alzheimer's disease in a large sample of patients; and (ii) to investigate the pattern of glucose metabolism as a function of dementia severity in early onset versus late onset Alzheimer's disease, using a statistical parametric mapping (SPM) analysis. Subjects consisted of four groups: 74 patients with early onset Alzheimer's disease, 46 patients with late onset of the disease, and two control groups age matched to each patient group. All the subjects underwent 2-[(18)F]fluoro-2-deoxy-d-glucose (FDG)-PET under the same scanning conditions. Severity of dementia was rated with the Clincial Dementia Rating (CDR). Voxel-based SPM99 was used for statistical analyses. Overall glucose hypometabolism of early onset Alzheimer's disease patients was much greater in magnitude and extent than that of late onset patients, though both groups were similar in dementia severity: the early onset group showed more severe hypometabolism in parietal, frontal and subcortical (basal ganglia and thalamus) areas. When the decline of glucose metabolism was compared as a function of CDR stage, the slope was steeper in early onset than in late onset Alzheimer's disease. The rapid decline occurred at CDR 0.5-1 in the early onset group, whereas similar changes occurred at CDR 2-3 in the late onset group. The greater hypometabolism in early onset than in late onset patients is required to reach the same severity of dementia, probably reflecting greater functional reserve in younger than in older subjects. Alternatively, the metabolic decline curve suggests that the early onset patients may take a more rapid course in the reduction of glucose metabolism than the late onset patients. PMID- 15888537 TI - Physiological thresholds for irreversible tissue damage in contusional regions following traumatic brain injury. AB - Cerebral ischaemia appears to be an important mechanism of secondary neuronal injury in traumatic brain injury (TBI) and is an important predictor of outcome. To date, the thresholds of cerebral blood flow (CBF) and cerebral oxygen utilization (CMRO(2)) for irreversible tissue damage used in TBI studies have been adopted from experimental and clinical ischaemic stroke studies. Identification of irreversibly damaged tissue in the acute phase following TBI could have considerable therapeutic and prognostic implications. However, it is questionable whether stroke thresholds are applicable to TBI. Therefore, the aim of this study was to determine physiological thresholds for the development of irreversible tissue damage in contusional and pericontusional regions in TBI, and to determine the ability of such thresholds to accurately differentiate irreversibly damaged tissue. This study involved 14 patients with structural abnormalities on late-stage MRI, all of whom had been studied with (15)O PET within 72 h of TBI. Lesion regions of interest (ROI) and non-lesion ROIs were constructed on late-stage MRIs and applied to co-registered PET maps of CBF, CMRO(2) and oxygen extraction fraction (OEF). From the entire population of voxels in non-lesion ROIs, we determined thresholds for the development of irreversible tissue damage as the lower limit of the 95% confidence interval for CBF, CMRO(2) and OEF. To test the ability of a physiological variable to differentiate lesion and non-lesion tissue, we constructed probability curves, demonstrating the ability of a physiological variable to predict lesion and non lesion outcomes. The lower limits of the 95% confidence interval for CBF, CMRO(2) and OEF in non-lesion tissue were 15.0 ml/100 ml/min, 36.7 mumol/100 ml/min and 25.9% respectively. Voxels below these values were significantly more frequent in lesion tissue (all P < 0.005, Mann-Whitney U-test). However, a significant proportion of lesion voxels had values above these thresholds, so that definition of the full extent of irreversible tissue damage would not be possible based upon single physiological thresholds. We conclude that, in TBI, the threshold of CBF below which irreversible tissue damage consistently occurs differs from the classical CBF threshold for stroke (where similar methodology is used to define such thresholds). The CMRO(2) threshold is comparable to that reported in the stroke literature. At a voxel-based level, however (and in common with ischaemic stroke), the extent of irreversible tissue damage cannot be accurately predicted by early abnormalities of any single physiological variable. PMID- 15888538 TI - Treatment-responsive limbic encephalitis identified by neuropil antibodies: MRI and PET correlates. AB - We report seven patients, six from a single institution, who developed subacute limbic encephalitis initially considered of uncertain aetiology. Four patients presented with symptoms of hippocampal dysfunction (i.e. severe short-term memory loss) and three with extensive limbic dysfunction (i.e. confusion, seizures and suspected psychosis). Brain MRI and [(18)F]fluorodeoxyglucose (FDG)-PET complemented each other but did not overlap in 50% of the patients. Combining both tests, all patients had temporal lobe abnormalities, five with additional areas involved. In one patient, FDG hyperactivity in the brainstem that was normal on MRI correlated with central hypoventilation; in another case, hyperactivity in the cerebellum anticipated ataxia. All patients had abnormal CSF: six pleocytosis, six had increased protein concentration, and three of five examined had oligoclonal bands. A tumour was identified and removed in four patients (mediastinal teratoma, thymoma, thymic carcinoma and thyroid cancer) and not treated in one (ovarian teratoma). An immunohistochemical technique that facilitates the detection of antibodies to cell surface or synaptic proteins demonstrated that six patients had antibodies to the neuropil of hippocampus or cerebellum, and one to intraneuronal antigens. Only one of the neuropil antibodies corresponded to voltage-gated potassium channel (VGKC) antibodies; the other five (two with identical specificity) reacted with antigens concentrated in areas of high dendritic density or synaptic-enriched regions of the hippocampus or cerebellum. Preliminary characterization of these antigens indicates that they are diverse and expressed on the neuronal cell membrane and dendrites; they do not co-localize with VGKCs, but partially co-localize with spinophilin. A target autoantigen in one of the patients co-localizes with a cell surface protein involved in hippocampal dendritic development. All patients except the one with antibodies to intracellular antigens had dramatic clinical and neuroimaging responses to immunotherapy or tumour resection; two patients had neurological relapse and improved with immunotherapy. Overall, the phenotype associated with the novel neuropil antibodies includes dominant behavioural and psychiatric symptoms and seizures that often interfere with the evaluation of cognition and memory, and brain MRI or FDG-PET abnormalities less frequently restricted to the medial temporal lobes than in patients with classical paraneoplastic or VGKC antibodies. When compared with patients with VGKC antibodies, patients with these novel antibodies are more likely to have CSF inflammatory abnormalities and systemic tumours (teratoma and thymoma), and they do not develop SIADH-like hyponatraemia. Although most autoantigens await characterization, all share intense expression by the neuropil of hippocampus, with patterns of immunolabelling characteristic enough to suggest the diagnosis of these disorders and predict response to treatment. PMID- 15888539 TI - The clinical characteristics of headache in patients with pituitary tumours. AB - The clinical characteristics of 84 patients with pituitary tumour who had troublesome headache were investigated. The patients presented with chronic (46%) and episodic (30%) migraine, short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT; 5%), cluster headache (4%), hemicrania continua (1%) and primary stabbing headache (27%). It was not possible to classify the headache according to International Headache Society diagnostic criteria in six cases (7%). Cavernous sinus invasion was present in the minority of presentations (21%), but was present in two of three patients with cluster headache. SUNCT-like headache was only seen in patients with acromegaly and prolactinoma. Hypophysectomy improved headache in 49% and exacerbated headache in 15% of cases. Somatostatin analogues improved acromegaly associated headache in 64% of cases, although rebound headache was described in three patients. Dopamine agonists improved headache in 25% and exacerbated headache in 21% of cases. In certain cases, severe exacerbations in headache were observed with dopamine agonists. Headache appears to be a significant problem in pituitary disease and is associated with a range of headache phenotypes. The presenting phenotype is likely to be governed by a combination of factors, including tumour activity, relationship to the cavernous sinus and patient predisposition to headache. A proposed modification of the current classification of pituitary-associated headache is given. PMID- 15888540 TI - Failure to replicate previously reported genetic associations with sporadic temporal lobe epilepsy: where to from here? AB - Temporal lobe epilepsy (TLE), traditionally thought to develop largely due to environmental factors, has recently become the focus of association studies in an effort to determine genetic risk factors. Here we examine all previous claims of association of genetic polymorphisms with TLE by attempting replication in a cohort of 339 TLE patients of European origin. We also examine if these variants contribute to other types of epilepsy by examination in a larger cohort of 752 patients representing a range of different epilepsies. We fail to clearly replicate any of the previously reported associations and also fail to show a role for these variants in the development of other forms of epilepsy. Although our results cannot definitively rule out a role for these genes, they do suggest that most and perhaps all of the previous associations are false positives. As has been the experience with other diseases, these results highlight the importance of larger sample sizes and replication. In TLE, it appears that collaboration before publication is the best option to increase sample size sufficiently in the short term. These general principles are applicable to other studies undertaken for common complex diseases. PMID- 15888541 TI - Pathophysiological heterogeneity of conduction blocks in multifocal motor neuropathy. AB - The pathophysiological mechanisms responsible for conduction block in multifocal motor neuropathy (MMN) are still unclear. To clarify the physiological abnormalities at the site of the block, we tested the effects induced by polarizing direct currents on motor conduction along forearm nerves in 25 normal nerves (13 subjects), and at the site of conduction block in six nerves (five patients) with MMN. In healthy controls, whereas nerve depolarization failed to change the conditioned compound muscle action potential (CMAP), hyperpolarization elicited a significant, charge-dependent, decrease in the conditioned CMAP size. Hyperpolarization with 4 mC elicited CMAPs that were 86.76 +/- 5.22% (mean +/- SEM) of the control unconditioned response (P < 0.05). Analysis of individual MMN nerves showed that polarizing currents elicited markedly heterogeneous effects: depending on the nerve tested, depolarization or hyperpolarization in most cases significantly improved conduction along motor fibres across the conduction block. In three MMN nerves, pathophysiological abnormalities were consistent with a hyperpolarizing block, in two with a depolarizing block, and in one with a mixed block. Our observations indicate that the pathophysiological abnormalities at the site of conduction block in MMN may arise from depolarization or hyperpolarization, probably depending on the course of disease. PMID- 15888542 TI - Spindle pole organization in Drosophila S2 cells by dynein, abnormal spindle protein (Asp), and KLP10A. AB - Dynein is a critical mitotic motor whose inhibition causes defects in spindle pole organization and separation, chromosome congression or segregation, and anaphase spindle elongation, but results differ in different systems. We evaluated the functions of the dynein-dynactin complex by using RNA interference (RNAi)-mediated depletion of distinct subunits in Drosophila S2 cells. We observed a striking detachment of centrosomes from spindles, an increase in spindle length, and a loss of spindle pole focus. RNAi depletion of Ncd, another minus-end motor, produced disorganized spindles consisting of multiple disconnected mini-spindles, a different phenotype consistent with distinct pathways of spindle pole organization. Two candidate dynein-dependent spindle pole organizers also were investigated. RNAi depletion of the abnormal spindle protein, Asp, which localizes to focused poles of control spindles, produced a severe loss of spindle pole focus, whereas depletion of the pole-associated microtubule depolymerase KLP10A increased spindle microtubule density. Depletion of either protein produced long spindles. After RNAi depletion of dynein dynactin, we observed subtle but significant mislocalization of KLP10A and Asp, suggesting that dynein-dynactin, Asp, and KLP10A have complex interdependent functions in spindle pole focusing and centrosome attachment. These results extend recent findings from Xenopus extracts to Drosophila cultured cells and suggest that common pathways contribute to spindle pole organization and length determination. PMID- 15888543 TI - The mitotic cyclins Clb2p and Clb4p affect morphogenesis in Candida albicans. AB - The ability of Candida albicans to switch cellular morphologies is crucial for its ability to cause infection. Because the cell cycle machinery participates in Saccharomyces cerevisiae filamentous growth, we characterized in detail the two C. albicans B-type cyclins, CLB2 and CLB4, to better understand the molecular mechanisms that underlie the C. albicans morphogenic switch. Both Clb2p and Clb4p levels are cell cycle regulated, peaking at G2/M and declining before mitotic exit. On hyphal induction, the accumulation of the G1 cyclin Cln1p was prolonged, whereas the accumulation of both Clb proteins was delayed when compared with yeast form cells, indicating that CLB2 and CLB4 are differentially regulated in the two morphologies and that the dynamics of cyclin appearance differs between yeast and hyphal forms of growth. Clb2p-depleted cells were inviable and arrested with hyper-elongated projections containing two nuclei, suggesting that Clb2p is not required for entry into mitosis. Unlike Clb2p-depleted cells, Clb4p-depleted cells were viable and formed constitutive pseudohyphae. Clb proteins lacking destruction box domains blocked cell cycle progression resulting in the formation of long projections, indicating that both Clb2p and Clb4p must be degraded before mitotic exit. In addition, overexpression of either B-type cyclin reduced the extent of filamentous growth. Taken together, these data indicate that Clb2p and Clb4p regulate C. albicans morphogenesis by negatively regulating polarized growth. PMID- 15888544 TI - The Golgi-associated protein GRASP65 regulates spindle dynamics and is essential for cell division. AB - At the onset of mitosis, the pericentriolar Golgi apparatus of mammalian cells is converted into small fragments, which are dispersed throughout the cytosol. The Golgi-associated protein GRASP65 is involved in this process. To address the role of GRASP65 in mitotic Golgi fragmentation, we depleted the protein from HeLa cells by RNAi. In the absence of GRASP65, the number of cisternae per Golgi stack is reduced without affecting the overall organization of Golgi membranes and protein transport. GRASP65-depleted cells entered mitosis, but accumulated in metaphase with condensed chromatin and multiple aberrant spindles and eventually died. Although Centrin2 and g-tubulin were detected in two of the spindle poles, the other spindle poles contained g-tubulin, but not Centrin2. Furthermore, we provide evidence that the expression of the C-terminus of GRASP65 interferes with entry of cells into mitosis. Our results suggest the requirement for GRASP65 in the regulation of spindle dynamics rather than a direct role in the stacking of Golgi cisternae. This novel function is in addition to the previously established negative role of GRASP65 at the G2/M transition, which is mediated by its C terminus. PMID- 15888545 TI - Nuclear import of TFIIB is mediated by Kap114p, a karyopherin with multiple cargo binding domains. AB - Nuclear import and export is mediated by an evolutionarily conserved family of soluble transport factors, the karyopherins (referred to as importins and exportins). The yeast karyopherin Kap114p has previously been shown to import histones H2A and H2B, Nap1p, and a component of the preinitiation complex (PIC), TBP. Using a proteomic approach, we have identified several potentially new cargoes for Kap114p. These cargoes include another PIC component, the general transcription factor IIB or Sua7p, which interacted directly with Kap114p. Consistent with its role as a Sua7p import factor, deletion of KAP114 led to specific mislocalization of Sua7p to the cytoplasm. An interaction between Sua7p and TBP was also detected in cytosol, raising the possibility that both Sua7p and TBP can be coimported by Kap114p. We have also shown that Kap114p possesses multiple overlapping binding sites for its partners, Sua7p, Nap1p, and H2A and H2B, as well as RanGTP and nucleoporins. In addition, we have assembled an in vitro complex containing Sua7p, Nap1p, and histones H2A and H2B, suggesting that this Kap may import several proteins simultaneously. The import of more than one cargo at a time would increase the efficiency of each import cycle and may allow the regulation of coimported cargoes. PMID- 15888546 TI - Specific features of neuronal size and shape are regulated by tropomyosin isoforms. AB - Spatially distinct populations of microfilaments, characterized by different tropomyosin (Tm) isoforms, are present within a neuron. To investigate the impact of altered tropomyosin isoform expression on neuronal morphogenesis, embryonic cortical neurons from transgenic mice expressing the isoforms Tm3 and Tm5NM1, under the control of the beta-actin promoter, were cultured in vitro. Exogenously expressed Tm isoforms sorted to different subcellular compartments with Tm5NM1 enriched in filopodia and growth cones, whereas the Tm3 was more broadly localized. The Tm5NM1 neurons displayed significantly enlarged growth cones accompanied by an increase in the number of dendrites and axonal branching. In contrast, Tm3 neurons displayed inhibition of neurite outgrowth. Recruitment of Tm5a and myosin IIB was observed in the peripheral region of a significant number of Tm5NM1 growth cones. We propose that enrichment of myosin IIB increases filament stability, leading to the enlarged growth cones. Our observations support a role for different tropomyosin isoforms in regulating interactions with myosin and thereby regulating morphology in specific intracellular compartments. PMID- 15888547 TI - Role of SHPS-1 in the regulation of insulin-like growth factor I-stimulated Shc and mitogen-activated protein kinase activation in vascular smooth muscle cells. AB - Insulin-like growth factor I (IGF-I) stimulates smooth muscle cell (SMC) proliferation, and the mitogen-activated protein kinase (MAPK) pathway plays an important role in mediating IGF-I-induced mitogenic signaling. Our prior studies have shown that recruitment of Src homology 2 domain tyrosine phosphatase (SHP-2) to the membrane scaffolding protein Src homology 2 domain-containing protein tyrosine phosphatase substrate-1 (SHPS-1) is required for IGF-I-dependent MAPK activation. The current studies were undertaken to define the upstream signaling components that are required for IGF-I-stimulated MAPK activation and the role of SHPS-1 in regulating this process. The results show that IGF-I-induced Shc phosphorylation and its subsequent binding to Grb2 is required for sustained phosphorylation of MAPK and increased cell proliferation in SMCs. Furthermore, for Shc to be phosphorylated in response to IGF-I requires that Shc must associate with SHPS-1 and this association is mediated in part by SHP-2. Preincubation of cells with a peptide that contains a phospho-tyrosine binding motif sequence derived from SHPS-1 inhibited IGF-I-stimulated SHP-2 transfer to SHPS-1, the association of Shc with SHPS-1, and IGF-I-dependent Shc phosphorylation. Expression of an SHPS-1 mutant that did not bind to Shc or SHP-2 resulted in decreased Shc and MAPK phosphorylation in response to IGF-I. In addition, SMCs expressing a mutant form of the beta3 subunit of the alphaVbeta3, which results in impairment of SHP-2 transfer to SHPS-1, also showed attenuated IGF-I-dependent Shc and MAPK phosphorylation. Further analysis showed that Shc and SHP-2 can be coimmunoprecipitated after IGF-I stimulation. A cell-permeable peptide that contained a polyproline sequence from Shc selectively inhibited Shc/SHP-2 association and impaired Shc but not SHP-2 binding to SHPS-1. Exposure to this peptide also inhibited IGF-I-stimulated Shc and MAPK phosphorylation. Cells expressing a mutant form of Shc with the four prolines substituted with alanines showed no Shc/SHPS-1 association in response to IGF-I. We conclude that SHPS-1 functions as an anchor protein that recruits both Shc and SHP-2 and that their recruitment is necessary for IGF-I-dependent Shc phosphorylation, which is required for an optimal mitogenic response in SMCs. PMID- 15888548 TI - ELKS, a protein structurally related to the active zone-associated protein CAST, is expressed in pancreatic beta cells and functions in insulin exocytosis: interaction of ELKS with exocytotic machinery analyzed by total internal reflection fluorescence microscopy. AB - The cytomatrix at the active zone (CAZ) has been implicated in defining the site of Ca2+-dependent exocytosis of neurotransmitters. Here, we demonstrate the expression and function of ELKS, a protein structurally related to the CAZ protein CAST, in insulin exocytosis. The results of confocal and immunoelectron microscopic analysis showed that ELKS is present in pancreatic beta cells and is localized close to insulin granules docked on the plasma membrane-facing blood vessels. Total internal reflection fluorescence microscopy imaging in insulin producing clonal cells revealed that the ELKS clusters are less dense and unevenly distributed than syntaxin 1 clusters, which are enriched in the plasma membrane. Most of the ELKS clusters were on the docking sites of insulin granules that were colocalized with syntaxin 1 clusters. Total internal reflection fluorescence images of single-granule motion showed that the fusion events of insulin granules mostly occurred on the ELKS cluster, where repeated fusion was sometimes observed. When the Bassoon-binding region of ELKS was introduced into the cells, the docking and fusion of insulin granules were markedly reduced. Moreover, attenuation of ELKS expression by small interfering RNA reduced the glucose-evoked insulin release. These data suggest that the CAZ-related protein ELKS functions in insulin exocytosis from pancreatic beta cells. PMID- 15888549 TI - Regulation of IkappaB kinase and NF-kappaB in contracting adult rat skeletal muscle. AB - Nuclear factor-kappaB (NF-kappaB) is a transcription factor with important roles in regulating innate immune and inflammatory responses. NF-kappaB is activated through the phosphorylation of its inhibitor, IkappaB, by the IkappaB kinase (IKK) complex. Physical exercise elicits changes in skeletal muscle gene expression, yet signaling cascades and transcription factors involved remain largely unknown. To determine whether NF-kappaB signaling is regulated by exercise in vivo, rats were run on a motorized treadmill for 5-60 min. Exercise resulted in up to twofold increases in IKKalpha/beta phosphorylation in the soleus and red gastrocnemius muscles throughout the time course studied. In red gastrocnemius muscles, NF-kappaB activity increased 50% 1-3 h after 60 min of treadmill exercise, returning to baseline by 5 h. Contraction of isolated extensor digitorum longus muscles in vitro increased IKKalpha/beta phosphorylation sevenfold and this was accompanied by a parallel increase in IkappaBalpha phosphorylation. Additional kinases that are activated by exercise include p38, extracellular-signal regulated protein kinase (ERK), and AMP activated protein kinase (AMPK). Inhibitors of p38 (SB-203580) and ERK (U-0126) blunted contraction-mediated IKK phosphorylation by 39 +/- 4% (P = 0.06) and 35 +/- 10% (P = 0.09), respectively, and in combination by 76 +/- 5% (P < 0.05), suggesting that these kinases might influence the activation of IKK and NF-kappaB during exercise. In contrast, 5-aminoimidazole-4-carboxamide-1-beta-D ribofuranoside, an activator of AMPK, had no effect on either IKK or NF-kappaB activity. In conclusion, acute submaximal exercise transiently stimulates NF kappaB signaling in skeletal muscle. This activation is a local event because it can occur in the absence of exercise-derived systemic factors. PMID- 15888550 TI - Distinct and sequential upregulation of genes regulating cell growth and cell cycle progression during hepatic ischemia-reperfusion injury. AB - Ischemia-reperfusion injury (IRI) in liver and other organs is manifested as an injury phase followed by recovery and resolution. Control of cell growth and proliferation is essential for recovery from the injury. We examined the expression of three related regulators of cell cycle progression in liver IRI: spermidine/spermine N-acetyltransferase (SSAT), p21 (a cyclin-dependent kinase inhibitor), and stathmin. Mice were subjected to hepatic IRI, and liver tissues were harvested at timed intervals. The expression of SSAT, the rate-limiting enzyme in the polyamine catabolic pathway, had increased fivefold 6 h after IRI and correlated with increased putrescine levels in the liver, consistent with increased SSAT enzymatic activity in IRI. The expression of p21, which is transactivated by p53, was undetectable in sham-operated animals but was heavily induced at 12 and 24 h of reperfusion and declined to undetectable baseline levels at 72 h of reperfusion. The interaction of the polyamine pathway with the p53-p21 pathway was shown in vitro, where activation of SSAT with polyamine analog or the addition of putrescine to cultured hepatocytes induced the expression of p53 and p21 and decreased cell viability. The expression of stathmin, which is under negative transcriptional regulation by p21 and controls cell proliferation and progression through mitosis, remained undetectable at 6, 12, and 24 h of reperfusion and was progressively and heavily induced at 48 and 72 h of reperfusion. Double-immunofluorescence labeling with antibodies against stathmin and PCNA, a marker of cell proliferation, demonstrated colocalization of stathmin and PCNA at 48 and 72 h of reperfusion in hepatocytes, indicating the initiation of cell proliferation. The distinct and sequential upregulation of SSAT, p21, and stathmin, along with biochemical activation of the polyamine catabolic pathway in IRI in vivo and the demonstration of p53-p21 upregulation by SSAT and putrescine in vitro, points to the important role of regulators of cell growth and cell cycle progression in the pathophysiology and/or recovery in liver IRI. The data further suggest that SSAT may play a role in the initiation of injury, whereas p21 and stathmin may be involved in the resolution and recovery after liver IRI. PMID- 15888551 TI - Activation of NHE3 by dexamethasone requires phosphorylation of NHE3 at Ser663 by SGK1. AB - Glucocorticoids stimulate Na+ absorption by activation of the epithelial Na+/H+ exchanger NHE3 in the kidney and intestine. It has been thought that glucocorticoid-induced activation of NHE3 is solely dependent on transcriptional induction of the NHE3 gene. While the transcriptional regulation remains an essential part of the chronic effect of glucocorticoids, a previous study by us identified the serum- and glucocorticoid-inducible kinase 1 (SGK1) as an important component of the activation of NHE3 by glucocorticoids. In this work, we have demonstrated phosphorylation of NHE3 by SGK1 as the key mechanism for the stimulation of the transport activity by glucocorticoids. By using in vitro SGK1 kinase assay and site-directed mutagenesis, we have identified Ser663 of NHE3 to be the major site of phosphorylation by SGK1. Ser663 is invariantly conserved in all NHE3 proteins from several species, and the mutation of Ser663 to Ala blocks the effect of dexamethasone, demonstrating the importance of phosphorylation at Ser663. We also show that phosphorylation of NHE3 precedes the changes in NHE3 activity, and the increased activity is associated with an increased amount of NHE3 proteins in the surface membrane. These data reveal that dexamethasone activates NHE3 activity by phosphorylating the NHE3 protein, which initiates trafficking of the protein into the plasma membrane. PMID- 15888552 TI - Wnt/beta-catenin signaling activates growth-control genes during overload-induced skeletal muscle hypertrophy. AB - Beta-catenin is a transcriptional activator shown to regulate the embryonic, postnatal, and oncogenic growth of many tissues. In most research to date, beta catenin activation has been the unique downstream function of the Wnt signaling pathway. However, in the heart, a Wnt-independent mechanism involving Akt mediated phosphorylation of glycogen synthase kinase (GSK)-3beta was recently shown to activate beta-catenin and regulate cardiomyocyte growth. In this study, results have identified the activation of the Wnt/beta-catenin pathway during hypertrophy of mechanically overloaded skeletal muscle. Significant increases in beta-catenin were determined during skeletal muscle hypertrophy. In addition, the Wnt receptor, mFrizzled (mFzd)-1, the signaling mediator disheveled-1, and the transcriptional co-activator, lymphocyte enhancement factor (Lef)-1, are all increased during hypertrophy of the overloaded mouse plantaris muscle. Experiments also determined an increased association between GSK-3beta and the inhibitory frequently rearranged in advanced T cell-1 protein with no increase in GSK-3beta phosphorylation (Ser9). Finally, skeletal muscle overload resulted in increased nuclear beta-catenin/Lef-1 expression and induction of the transcriptional targets c-Myc, cyclin D1, and paired-like homeodomain transcription factor 2. Thus this study provides the first evidence that the Wnt signaling pathway induces beta-catenin/Lef-1 activation of growth-control genes during overload induced skeletal muscle hypertrophy. PMID- 15888553 TI - Effects of the JNK inhibitor anthra[1,9-cd]pyrazol-6(2H)-one (SP-600125) on soluble guanylyl cyclase alpha1 gene regulation and cGMP synthesis. AB - The decreased expression of the nitric oxide (NO) receptor, soluble guanylyl cyclase (sGC), occurs in response to multiple stimuli in vivo and in cell culture and correlates with various disease states such as hypertension, inflammation, and neurodegenerative disorders. The ability to understand and modulate sGC expression and cGMP levels in any of these conditions could be a valuable therapeutic tool. We demonstrate herein that the c-Jun NH2-terminal kinase JNK II inhibitor anthra[1,9-cd]pyrazol-6(2H)-one (SP-600125) completely blocked the decreased expression of sGCalpha1-subunit mRNA by nerve growth factor (NGF) in PC12 cells. Inhibitors of the ERK and p38 MAPK pathways, PD-98059 and SB-203580, had no effect. SP-600125 also inhibited the NGF-mediated decrease in the expression of sGCalpha1 protein as well as sGC activity in PC12 cells. Other experiments revealed that decreased sGCalpha1 mRNA expression through a cAMP mediated pathway, using forskolin, was not blocked by SP-600125. We also demonstrate that TNF-alpha/IL-1beta stimulation of rat fetal lung (RFL-6) fibroblast cells resulted in sGCalpha1 mRNA inhibition, which was blocked by SP 600125. Expression of a constitutively active JNKK2-JNK1 fusion protein in RFL-6 cells caused endogenous sGCalpha1 mRNA levels to decrease, while a constitutively active ERK2 protein had no effect. Collectively, these data demonstrate that SP 600125 may influence the intracellular levels of the sGCalpha1-subunit in certain cell types and may implicate a role for c-Jun kinase in the regulation of sGCalpha1 expression. PMID- 15888554 TI - Characteristics of new Staphylococcus aureus-RBC adhesion mechanism independent of fibrinogen and IgG under hydrodynamic shear conditions. AB - Staphylococcus aureus infection begins when bacterial cells circulating in blood adhere to components of the extracellular matrix or endothelial cells of the host and initiate colonization. S. aureus is known to exhibit extensive interactions with platelets. S. aureus is also known to bind to red blood cells (RBCs) in the presence of plasma proteins, such as fibrinogen and IgG. Herein we report a new binding mechanism of S. aureus to RBC independent of those plasma proteins. To characterize the new adhesion mechanism, we experimentally examine the binding kinetics and molecular constituents mediating the new adhesive interactions between S. aureus and RBCs under defined shear conditions. The results demonstrate that the receptors for fibrinogen (clumping factor A) and IgG (protein A) of S. aureus are not involved in the adhesion. S. aureus binds to RBCs with maximal adhesion at the shear rate 100 s(-1) and decreasing adhesion with increasing shear. The heteroaggregates formed after shear are stable when subjected to the shear rate 2,000 s(-1), indicating that intercellular contact time rather than shear forces controls the adhesion at high shear. S. aureus binding to RBC requires plasma, and 10% plasma is sufficient for maximal adhesion. Plasma proteins involved in the cell-cell adhesion, such as fibrinogen, fibronectin, von Willebrand factor, IgG, thrombospondin, laminin, and vitronectin are not involved in the observed adhesion. The extent of heteroaggregation is dramatically reduced on RBC treatment with trypsin, chymotrypsin, or neuraminidase, suggesting that the receptor(s) mediating the heteroaggregation process is a sialylated glycoprotein on RBC surface. Adhesion is divalent cation dependent and also blocked by heparin. This work demonstrates a new mechanism of S. aureus-RBC binding under hydrodynamic shear conditions via unknown RBC sialoglycoprotein(s). The binding requires plasma protein(s) other than fibrinogen or IgG and does not involve the S. aureus adhesins clumping factor A or protein A. PMID- 15888555 TI - The Poggendorff illusion explained by natural scene geometry. AB - One of the most intriguing of the many discrepancies between perceived spatial relationships and the physical structure of visual stimuli is the Poggendorff illusion, when an obliquely oriented line that is interrupted no longer appears collinear. Although many different theories have been proposed to explain this effect, there has been no consensus about its cause. Here, we use a database of range images (i.e., images that include the distance from the image plane of every pixel in the scene) to show that the probability distribution of the possible locations of line segments across an interval in natural environments can fully account for all of the behavior of this otherwise puzzling phenomenon. PMID- 15888556 TI - Cross-reactivity and conformational multiplicity of an anti-polycyclic aromatic hydrocarbon mAb. AB - Cross-reactivity and multispecific functionality of antibodies play a central role in the immune system. The Ab's promiscuity is attributed to structural flexibility and conformational multiplicity of its binding sites governed by the rearrangement of hydrogen bonding centers. However, antibodies whose recognition and binding rely on less directional hydrophobic interactions might follow different interaction pathways. We investigated interaction of anti-polycyclic aromatic hydrocarbon mAb with two biologically important cross-reactants, pyrene and benzo(a)pyrene. Complex formation was characterized by means of low temperature laser-induced fluorescence spectroscopy in both low- and high resolution fluorescence line-narrowing (FLN) modes. It is shown that the FLN spectroscopy can identify various haptens cross-reacted with an Ab, as well as simultaneously differentiate between free and immunocomplexed haptens. In addition, our results suggest an interesting case of an Ab binding a particular cross-reactant by adopting two distinct conformations of its binding sites. The existence of the multiple conformations for anti-polycyclic aromatic hydrocarbon mAb that are trapped at low temperature can be rationalized through the existing models for Ab binding. Finally, as revealed by FLN spectra of immunocomplexed chromophores, pi-pi interactions, rather than hydrogen bonding, play the central role in complex formation. PMID- 15888557 TI - American Society of Nephrology Renal Research Report. AB - In the spring of 2004, the Board of Advisors and the Council of the American Society of Nephrology believed it necessary to conduct a series of research retreats to steer priorities appropriately in an era of limited resources. In this regard, retreats were conducted by five working groups in areas that were identified to require distinct attention: acute renal failure, diabetic nephropathy, hypertension, transplantation, and uremic cardiovascular toxicity. The goal of each retreat was to join experts, both within and outside the renal community, to identify areas of basic science and clinical research that should receive highest priority in the next five years. The five retreat summaries with their individual listings of research priorities allow for the distillation of three overriding recommendations that strongly emanate from them: Continued support and expansion of investigator initiated research projects. In each of the five subjects on which this report is focused, there are areas of investigation that require the support of investigator-initiated projects if ultimately progress is to be made in the understanding of the basic mechanisms that underlie the diseases processes on which we want to have an impact in the next decade. It is recommended that there be an expansion of support for research in the areas highlighted in this report that lend themselves to this mechanism of funding by encouraging applications with appropriate program announcements and requests for proposals. In addition to vigorous support for RO1 grants, continued funding of Concept Development and R21/R33 grants is essential to support development of investigator-initiated clinical studies in these areas of high priority. Support for the development of a collaborative research infrastructure. The reader of this article cannot but be impressed by the common theme that independently emerged from each report regarding the urgent need to develop an infrastructure for kidney research. This infrastructure requires the development of core facilities for the centralized processing of biologic materials (genomics, proteomics, and metabolomics), in vivo imaging, development and distribution of antibodies and other molecular reagents, development and distribution and phenotyping of mouse models, and perhaps others. These need to be complemented with core bioinformatics centers that collect and analyze data and finally with a network of clinical study coordinating centers. Expansion of kidney research infrastructure can be achieved by vigorous funding of a program of kidney research core centers. Specifically, we propose that the number of kidney centers be increased with the goal of providing core facilities to support collaborative research on a local, regional, and national level. It should be emphasized that such a program of competitively reviewed kidney core centers would facilitate investigator-initiated research in both laboratory and patient-oriented investigation. This approach is also very much in line with the collaborative research enterprise conceived in the National Institutes of Health's Road Map. Support programs that have an impact on the understanding of the relationship between renal and cardiovascular disease (CVD). It is now widely recognized that chronic kidney dysfunction is an important risk factor for the development of CVD. It therefore is not surprising that essentially every one of the retreat reports emphasizes the urgency to examine this relationship. It is recommended that the National Institute of Diabetes and Digestive and Kidney Diseases and the National Heart, Lung, and Blood Institute (NHLBI) work cooperatively to support both basic and clinical science projects that will shed light on the pathogenesis of this relationship and to support the exploration of interventions that can decrease cardiovascular events in patients with chronic kidney disease. Thus, we specifically propose that the NHLBI support investigator-initiated research (RO1, Concept Development, and R21/R33) grants in areas of kidney research with a direct relationship to CVD. Similarly, the NHLBI should work collaboratively with the National Institute of Diabetes and Digestive and Kidney Diseases to support the proposed program of kidney core research centers. This subject provides an excellent opportunity to foment a collaboration between two institutes, along the lines of the present-day overall philosophy of the National Institutes of Health. PMID- 15888558 TI - On the intraoperative molecular status of renal allografts after vascular reperfusion and clinical outcomes. AB - Many hypothesize that subtle inflammation and immune activity detected in the intraoperative period are linked to adverse postkidney transplant clinical outcomes. To this end, renal allografts were analyzed for expression of pro inflammatory, inflammation-induced adhesion molecules, immune activation as well as anti-apoptotic genes expressed 15 min after vascular reperfusion (zero-hour) to determine whether this analysis can aid in predicting the occurrence of delayed graft function (DGF), acute rejection (AR), and the quality of graft function at 6 mo. Intraoperative biopsies were obtained from 75 consecutively performed renal allografts in which consent was obtained 15 min after vascular reperfusion. These biopsies were analyzed by quantitative real-time PCR for transcription of 15 select genes and by standard histopathology. Posttransplant clinical outcomes were also analyzed in respect to intraoperative transcriptional profiles and clinical parameters available at the time of transplantation. This study demonstrates that a limited and hypothesis-driven PCR-based transcriptional profile of the zero-hour kidney biopsy predicts posttransplant clinical outcomes including DGF, early AR, and the quality of renal function 6 mo posttransplantation. For some clinical endpoints, the combined use of molecular analysis and established clinical indicators available at the time of transplantation further enhances the quality of prognosis. The transcriptional profiling data provide absolutely essential data to the predictive models, particularly with respect to AR and renal function 6 mo posttransplantation. PMID- 15888559 TI - Recurrence of membranoproliferative glomerulonephritis type II in renal allografts: The North American Pediatric Renal Transplant Cooperative Study experience. AB - Membranoproliferative glomerulonephritis type II (MPGN II) is an uncommon form of complement-dependent acquired renal disease. Although it has been recognized since the 1970s that MPGN II recurs almost universally in renal transplants, data regarding the long-term consequences of disease recurrence are limited. Therefore, a retrospective comparative analysis of 75 patients with MPGN II contained in the North American Pediatric Renal Transplant Cooperative Study transplantation database was performed. Five-year graft survival for patients with MPGN II was significantly worse (50.0 +/- 7.5%) compared with the database as a whole (74.3 +/- 0.6%; P < 0.001). Living related donor organs had a significantly better 5-yr survival (65.9 +/- 10.7%) compared with cadaveric donor organs (34.1 +/- 9.8%; P = 0.004). The primary cause of graft failure in 11 (14.7%) patients was recurrent disease. Supplemental surveys were obtained on 29 (38%) of 75 patients. Analysis of these data indicated that recurrent disease occurred in 12 (67%) of the 18 patients with posttransplantation biopsies. Although there was no correlation between pretransplantation presentation, pre- or posttransplantation C3 levels, and either disease recurrence or graft failure, there was a strong association between heavy proteinuria and disease recurrence. The presence of glomerular crescents in allograft biopsies had a significant negative correlation with graft survival. At last follow-up, patients with recurrent disease had significantly higher serum creatinine and qualitatively more proteinuria than patients without biopsy-proven disease. These data indicate that recurrent MPGN II has a significant negative impact on renal allograft function and survival. PMID- 15888560 TI - Subcutaneous ghrelin enhances acute food intake in malnourished patients who receive maintenance peritoneal dialysis: a randomized, placebo-controlled trial. AB - Anorexia and malnutrition confer significant morbidity and mortality to patients with end-stage kidney disease but are resistant to therapy. The aim of this study was to determine whether subcutaneous administration of ghrelin, an appetite stimulating gut hormone, could enhance food intake in patients who are receiving maintenance peritoneal dialysis and have evidence of malnutrition. The principal outcome measure was energy intake during a measured study meal. Secondary outcome measures were BP and heart rate and 3-d food intake after intervention. Nine peritoneal dialysis patients with mild to moderate malnutrition (mean serum albumin 28.6 +/- 5.0 g/L, total cholesterol 4.4 +/- 0.6 mmol/L, subjective global assessment score of 5.7 +/- 1.7) were given subcutaneous ghrelin (3.6 nmol/kg) and saline placebo in a randomized, double-blind, crossover protocol. Administration of subcutaneous ghrelin significantly increased the group mean absolute energy intake, compared with placebo, during the study meal (690 +/- 190 versus 440 +/- 250 kcal; P = 0.0062). When expressed as proportional energy increase for each individual, ghrelin administration resulted in immediate doubling of energy intake (204 +/- 120 versus 100%; P = 0.0319). Administration of ghrelin maintained a nonsignificant increase in energy intake over 24 h after intervention (2009 +/- 669 versus 1579 +/- 330 kcal) and was not followed by subsequent underswing (1790 +/- 370 versus 1670 +/- 530 and 1880 +/- 390 versus 1830 +/- 530 kcal on days 2 and 3, respectively). Ghrelin administration resulted in a significant fall in mean arterial BP (P = 0.0030 by ANOVA). There were no significant adverse events during the study. Subcutaneous ghrelin administration enhances short-term food intake in dialysis patients with mild to moderate malnutrition. PMID- 15888561 TI - Chronic kidney disease and cognitive impairment in the elderly: the health, aging, and body composition study. AB - Previous studies suggest a link between chronic kidney disease (CKD) and cognitive impairment. Whether the longitudinal course of cognitive impairment differs among people with or without CKD is unknown. Data collected in 3034 elderly individuals who participated in the Health, Aging, and Body Composition study were analyzed. Cognitive function was assessed with the Modified Mini Mental State Exam (3MS) at baseline and then 2 and 4 yr after baseline. Cognitive impairment was defined as a 3MS score <80 or a decline in 3MS >5 points after 2 or 4 yr of follow-up among participants with baseline 3MS scores > or =80. Participants with CKD, defined as an estimated GFR (eGFR) <60 ml/min per 1.73 m2, were further divided into two eGFR strata. Unadjusted mean baseline 3MS scores and mean declines in 3MS scores over 4 yr were significantly more pronounced for participants with lower baseline eGFR. More advanced stages of CKD were associated with an increased risk for cognitive impairment: Odds ratio (OR) 1.32 (95% confidence interval [CI] 1.03 to 1.69) and OR 2.43 (95% CI, 1.38 to 4.29) for eGFR 45 to 59 ml/min per 1.73 m2 and <45 ml/min per 1.73 m2, respectively, adjusted for case mix, baseline 3MS scores, and other potential confounders. CKD is associated with an increased risk for cognitive impairment in the elderly that cannot be fully explained by other well-established risk factors. Studies aimed at understanding the mechanism(s) responsible for cognitive impairment in CKD and efforts to interrupt this decline are warranted. PMID- 15888562 TI - Vasopressin-V2 receptor stimulation reduces sodium excretion in healthy humans. AB - In addition to its effect on water permeability, vasopressin, through its V2 receptors (AVPR2), stimulates Na reabsorption in the collecting duct by increasing the activity of the amiloride-sensitive sodium channel ENaC. This study evaluated whether dDAVP (a potent AVPR2 agonist) reduces sodium excretion in healthy humans (n = 6) and in patients with central (C; n = 2) or nephrogenic (N) diabetes insipidus (DI) as a result of mutations of either the aquaporin 2 gene (AQP2; n = 3) or AVPR2 (n = 10). dDAVP was infused intravenously (0.3 microg/kg body wt in 20 min), and urine was collected for 60 min before (basal) and 150 min after the infusion. dDAVP markedly reduced both urine flow rate and sodium excretion in healthy individuals. A reduction in sodium excretion was also observed in CDI and NDI-AQP2 patients but not in NDI-AVPR2 patients. The magnitude of the fall in sodium excretion correlated with the rise in urine osmolality and the fall in urine output but not with the simultaneously observed fall in mean BP. These results suggest that the dDAVP-induced antinatriuresis is due to a direct V2 receptor-dependent stimulation of sodium reabsorption in the collecting duct and is not secondary to a hemodynamic effect. In conclusion, this study reveals a potent V2-dependent antinatriuretic effect of vasopressin in humans. The possibility that an inappropriate stimulation of ENaC by vasopressin might lead to significant sodium retention in chronic situations remains to be determined. PMID- 15888563 TI - IL-12p40 and IL-18 in crescentic glomerulonephritis: IL-12p40 is the key Th1 defining cytokine chain, whereas IL-18 promotes local inflammation and leukocyte recruitment. AB - Experimental crescentic glomerulonephritis (GN) is characterized by T helper 1 (Th1) directed nephritogenic immune responses and cell-mediated glomerular injury. IL-12p40, the common cytokine chain for both IL-12 and IL-23, is important in the generation and potentially the maintenance of Th1 responses, whereas IL-18 is a co-factor for Th1 responses that may have systemic and local proinflammatory effects. For testing the hypothesis that both endogenous IL-12p40 and endogenous IL-18 play pathogenetic roles in crescentic GN, accelerated anti glomerular basement membrane GN was induced in mice genetically deficient in IL 12p40 (IL-12p40-/-), IL-18 (IL-18-/-), or both IL-12p40 and IL-18 (IL-12p40-/-IL 18-/-). Compared with wild-type C57BL/6 mice, IL-12p40-/- mice failed to make a nephritogenic Th1 response and developed markedly reduced crescent formation and renal leukocytic infiltration, despite renal production of chemoattractants and adhesion molecules. IL-18-/- mice developed an intact antigen-specific systemic Th1 response, a similar degree of crescent formation, but fewer glomeruli affected by other severe histologic changes and fewer leukocytes in glomeruli and interstitium. IL-18 was expressed within diseased kidneys. Local production of TNF, IL-1beta, IFN-gamma, CCL3 (MIP-1alpha), and CCL4 (MIP-1beta) was reduced in IL-18-/- mice, demonstrating a local proinflammatory role for IL-18. Combined deletion of IL-12p40 and IL-18 did not result in synergistic effects. Consistent with the hypothesis that inflammation leads to fibrosis, all three groups of deficient mice expressed lower levels of intrarenal TGF-beta1 and/or alpha1(I) procollagen mRNA. These studies demonstrate that in severe experimental crescentic GN, IL-12p40 is the key Th1-defining cytokine chain, whereas IL-18 has local proinflammatory roles. PMID- 15888564 TI - Increased osteoblastic activity and expression of receptor activator of NF-kappaB ligand in nonuremic nephrotic syndrome. AB - Patients with nephrotic syndrome (NS), even with normal GFR, often display altered mineral homeostasis and abnormal bone histology. However, the latter, mostly osteomalacia and increased bone resorption, cannot be readily explained by the prevalent concentrations of parathyroid hormone and vitamin D metabolites. The transmembrane receptor activator of NF-kappaB ligand (RANKL) of osteoblasts is essential for osteoclast formation and differentiation. Osteoblasts activity and the expression of RANKL were tested in cultures of normal human osteoblasts with sera obtained from patients with NS and normal GFR (129 +/- 26 ml/min per 1.73 m2) during relapse and remission of their NS. Osteoblasts that were cultured in vitro with sera during relapse displayed elevated concentrations of alkaline phosphatase (AP) and increased expression of RANKL. By contrast, during remission, AP concentrations were significantly lower (P < 0.05) and RANKL expression notably attenuated or absent. AP correlated with the proteinuria (r = 0.5, P < 0.05) and was not significantly affected by the therapeutic administration of corticosteroids. Whereas parathyroid hormone levels were normal (35 +/- 21 pg/ml), the serum markers of bone formation (osteocalcin and bone specific alkaline phosphatase) were lower during relapse compared with remission. Thus, sera from patients with NS and normal GFR stimulate the activity of osteoblasts and upregulate their expression of RANKL. These alterations, more prominent during clinically active NS, are transient and reversible upon remission. These disturbances of bone biology may play an important pathogenic role in the abnormal bone histology observed in patients with NS even before a decline in GFR occurs. PMID- 15888565 TI - De novo Uroplakin IIIa heterozygous mutations cause human renal adysplasia leading to severe kidney failure. AB - Human renal adysplasia usually occurs sporadically, and bilateral disease is the most common cause of childhood end-stage renal failure, a condition that is lethal without intervention using dialysis or transplantation. De novo heterozygous mutations in Uroplakin IIIa (UPIIIa) are reported in four of 17 children with kidney failure caused by renal adysplasia in the absence of an overt urinary tract obstruction. One girl and one boy in unrelated kindreds had a missense mutation at a CpG dinucleotide in the cytoplasmic domain of UPIIIa (Pro273Leu), both of whom had severe vesicoureteric reflux, and the girl had persistent cloaca; two other patients had de novo mutations in the 3' UTR (963 T- >G; 1003 T-->C), and they had renal adysplasia in the absence of any other anomaly. The mutations were absent in all sets of parents and in siblings, none of whom had radiologic evidence of renal adysplasia, and mutations were absent in two panels of 192 ethnically matched control chromosomes. UPIIIa was expressed in nascent urothelia in ureter and renal pelvis of human embryos, and it is suggested that perturbed urothelial differentiation may generate human kidney malformations, perhaps by altering differentiation of adjacent smooth muscle cells such that the metanephros is exposed to a functional obstruction of urine flow. With advances in renal replacement therapy, children with renal failure, who would otherwise have died, are surviving to adulthood. Therefore, although the mechanisms of action of the UPIIIa mutations have yet to be determined, these findings have important implications regarding genetic counseling of affected individuals who reach reproductive age. PMID- 15888566 TI - Involvement of renal progenitor tubular cells in epithelial-to-mesenchymal transition in fibrotic rat kidneys. AB - Renal progenitor tubular cells (label-retaining cells [LRC]) were recently identified in normal kidneys by in vivo bromodeoxyuridine (BrdU) labeling. This study was conducted to examine the behavior of LRC in renal fibrosis. BrdU was injected intraperitoneally into normal rats daily for 7 d. After a 2-wk chase period, unilateral ureteral obstruction (UUO) was induced in these rats. In normal and contralateral kidneys, LRC were observed scattering among tubular epithelial cells. After UUO, the number of the LRC significantly increased, and most of them were positive for proliferating cell nuclear antigen (PCNA). In contrast, PCNA+ cells lacking BrdU label were rarely observed. It is interesting that LRC were detected not only in tubules but also in the interstitium after UUO. Laminin staining showed that a number of the LRC were adjacent to the destroyed tubular basement membrane. Some tubules, including LRC, lost the expression of E-cadherin after UUO. A large number of cell populations expressed vimentin, heat shock protein 47, or alpha-smooth muscle actin in the UUO kidneys, and each population contained LRC. None of the LRC was positive for these fibroblastic markers in contralateral kidneys. When renal tubules from BrdU treated rats were cultured in the gel, some cells protruded from the periphery of the tubules and migrated into the gel. Most of these cells were BrdU+. Neither the total content of BrdU in the kidneys nor the number of LRC in bone marrow significantly changed after UUO. Collectively, these results suggest that LRC is a cell population that proliferates, migrates, and transdifferentiates into fibroblast-like cells during renal fibrosis. PMID- 15888567 TI - Enhanced intrarenal angiotensinogen contributes to early renal injury in spontaneously hypertensive rats. AB - This study was performed to determine whether augmented intrarenal angiotensinogen may contribute to the enhanced renal angiotensin II (Ang II) and associated tissue injury in spontaneously hypertensive rats (SHR). SHR and Wistar Kyoto rats (WKY) were maintained on a normal diet and killed at either 7 or 14 wk of age. Two groups of SHR received either an Ang II type 1 receptor blocker (ARB; olmesartan, 5 mg/d) or a triple therapy (hydralazine 7.5 mg/d, reserpine 0.15 mg/d, and hydrochlorothiazide 3 mg/d [HRH]) during weeks 7 through 14. Systolic BP and renal Ang II were significantly increased in SHR-14 (n = 8) compared with WKY-7, WKY-14, and SHR-7 (n = 8 each), and ARB treatment prevented these increases (n = 8). However, whereas HRH treatment prevented the development of hypertension in SHR, this combination therapy failed to decrease renal Ang II (n = 8). With the use of urine samples or fixed renal sections, renal injuries in rats were quantified in a semiautomated manner by the following six parameters: (1) urinary excretion rate of total protein, (2) glomerular sclerosis, (3) interstitial expansion, (4) and (5) numbers of monocytes/macrophages in interstitium or glomeruli, and (6) arterial proliferation. Angiotensinogen mRNA and protein levels in kidney cortex, measured by real-time reverse transcriptase PCR and Western blot analysis, respectively, and all six parameters of renal damage were changed in parallel, and ARB treatment also prevented these increases. However, HRH treatment failed to prevent these increases. These results indicate that SHR have enhanced intrarenal angiotensinogen production that contributes to increased Ang II levels leading to the development of hypertension and renal injury in this strain. PMID- 15888568 TI - Calcium-sensing receptor expression and parathyroid hormone secretion in hyperplastic parathyroid glands from humans. AB - In uremic patients, severe parathyroid hyperplasia is associated with reduced parathyroid calcium-sensing receptor (CaR) expression. Thus, in these patients, a high serum Ca concentration may be required to inhibit parathyroid hormone (PTH) secretion. This study compares the magnitude of reduction in CaR expression and the degree of the abnormality in Ca-regulated PTH release in vitro. A total of 50 glands from 23 hemodialysis patients with refractory hyperparathyroidism were studied. Tissue slices were incubated in vitro to evaluate (1) the PTH secretory output in a normal Ca concentration (1.25 mM) and (2) the PTH secretory response to high (1.5 mM) and low (0.6 mM) Ca concentration. Tissue aliquots were processed for determination of CaRmRNA expression. The results showed that, corrected for DNA, parathyroid tissue with lowest CaR expression secreted more PTH than that with relatively high CaR expression (146 +/- 23 versus 60 +/- 2 pg/microg DNA; P < 0.01). Furthermore, glands with low CaR expression demonstrated a blunted PTH secretory response to both the inhibitory effect of high Ca and the stimulatory effect of low Ca. The study also showed that the larger the gland, the lower the CaRmRNA expression. Thus, large parathyroid glands produce a large amount of PTH not only as a result of the increased gland size but also because the parathyroid tissue secretory output is increased. These abnormalities in PTH regulation are related to low CaR expression. PMID- 15888569 TI - Androgen receptor pathway in rats with autosomal dominant polycystic kidney disease. AB - Androgens have been implicated in mediating disease escalation in autosomal dominant polycystic kidney disease (ADPKD). Dihydrotestosterone (DHT), an agonist, and flutamide (FLT), an antagonist, were administered to Han:SPRD rats with ADPKD, and the role of androgen receptor (AR) abundance and activation on the enlargement and function of cystic kidneys was evaluated. Renal AR abundance determined by immunoblots in 8- to 10-wk-old Cy/+ male rats was naturally increased four-fold above that of littermate +/+ controls. In male Cy/+, castration decreased AR abundance below control +/+ by -89.4%, and AR expression within cyst mural epithelial cells was strikingly decreased. Castration of Cy/+ male rats also reduced the usual increases in kidney weight by -49.7%, kidney cyst area by -34.0%, and serum urea nitrogen by -72.8%; these indices were restored to precastration levels by DHT. In Cy/+ male rats, FLT administration reduced the increase in kidney weight by -27.6% and serum urea nitrogen by -53.7% and decreased the increment in AR expression by -84.2% in comparison with untreated +/+ controls. There was no effect of FLT in female rats. Immunoblot expression of phospho-extracellular signal-regulated kinase 1/2 (P-ERK) and B Raf, key intermediates in the mitogen-activated protein kinase pathway that are abnormally elevated in Cy/+, was unaffected by castration and/or administration of DHT or FLT. AR was not expressed in renal epithelial cell nuclei of androgen deficient rats but was displayed in most tubule and mural cyst cell nuclei of androgen-replete rats. In androgen-deficient Cy/+, 80.6% of renal epithelial cells that had entered the cell cycle (proliferating cell nuclear antigen positive) also expressed P-ERK. In androgen-replete rats, proliferating cell nuclear antigen-positive cells co-expressed AR (12.7%), P-ERK (36.4%), and P-ERK + AR (45.0%); 5.9% were probably stimulated by other mitogenic mechanisms. It is concluded that androgens potentiate renal cell proliferation and cyst enlargement through ERK1/2-dependent and ERK1/2-independent signaling mechanisms in Han:SPRD. It is suggested that the basal rate of cell proliferation is determined by ERK1/2 signaling to a major extent and that androgens have additive effects. PMID- 15888570 TI - Functional genomic characterization of delipidation elicited by trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) in a polygenic obese line of mice. AB - Gene expression was measured during t10c12-CLA-induced body fat reduction in a polygenic obese line of mice. Adult mice (n = 185) were allotted to a 2 x 2 factorial experiment consisting of either nonobese (ICR-control) or obese (M16 selected) mice fed a 7% fat, purified diet containing either 1% linoleic acid (LA) or 1% t10c12-CLA. Body weight (BW) by day 14 was 12% lower in CLA- compared with LA-fed mice (P < 0.0001). By day 14, t10c12-CLA reduced weights of epididymal, mesenteric, and brown adipose tissues, as a percentage of BW, in both lines by 30, 27, and 58%, respectively, and increased liver weight/BW by 34% (P < 0.0001). Total RNA was isolated and pooled (4 pools per tissue per day) from epididymal adipose (days 5 and 14) of the obese mice to analyze gene expression profiles using Agilent mouse oligo microarray slides representing > 20,000 genes. Numbers of genes differentially expressed by greater than or equal to twofold in epididymal adipose (days 5 and 14) were 29 and 125, respectively. It was concluded that, in adipose tissue, CLA increased expression of uncoupling proteins (1 and 2), carnitine palmitoyltransferase system, tumor necrosis factor alpha (P < 0.05), and caspase-3 but decreased expression of peroxisome proliferator-activated receptor-gamma, glucose transporter-4, perilipin, caveolin 1, adiponectin, resistin, and Bcl-2 (P < 0.01). In conclusion, this experiment has revealed candidate genes that will be useful in elucidating mechanisms of adipose delipidation. PMID- 15888571 TI - Physiology without borders. PMID- 15888572 TI - Transgenic and knockout mice in diabetes research: novel insights into pathophysiology, limitations, and perspectives. AB - Insulin resistance and type 2 diabetes are serious public health threats. Although enormous research efforts have been focused on the pathogenesis of these diseases, the underlying mechanisms remain only partly understood. Here we review mouse phenotypes resulting from inactivation of molecules responsible for the control of glucose metabolism that have led to novel insights into insulin action and the development of insulin resistance. In addition, more sophisticated strategies to manipulate genes in mice in the future are presented. PMID- 15888573 TI - Identity deception: not a crime for a stem cell. AB - Stem cell transdifferentiation in the adult organism is the most common and questioned mechanism of growth and repair. Recent data suggest that adult stem cells are capable of generating mature cells beyond their own tissue boundaries, a process called developmental plasticity. To date, the most versatile cell discovered is the bone marrow progenitor cell. PMID- 15888574 TI - Intracellular signaling: spatial and temporal control. AB - Cells integrate many inputs through complex networks of interacting signaling pathways. Systems approaches as well as computer-aided reductionist approaches attempt to "untangle the wires" and gain an intimate understanding of cells. But "understanding" any system is just the way that the human mind gains the ability to predict behavior. Computer simulations are an alternative way to achieve this goal--quite possibly the only way for complex systems. We have new tools to probe large sets of unknown interactions, and we have amassed enough detailed information to quantitatively describe many functional modules. Cell physiology has passed the threshold: the time to begin modeling is now. PMID- 15888575 TI - Development of structures and transport functions in the mouse placenta. AB - The placenta is essential for sustaining the growth of the fetus during gestation, and defects in its function result in fetal growth restriction or, if more severe, fetal death. Several molecular pathways have been identified that are essential for development of the placenta, and mouse mutants offer new insights into the cell biology of placental development and physiology of nutrient transport. PMID- 15888576 TI - Physiological mechanisms of tumor-cell invasion and migration. AB - Recent advances in understanding the complex biology of the microenvironment that underlies tumor invasion and migration have revealed novel and promising therapeutic targets. Pharmacological blockade of intra- and extracellular signaling events that regulate migration and survival of multiple cell types may disrupt the host-tumor conspiracy that allows escape from normal developmental regulation. PMID- 15888577 TI - Synaptic depression as a timing device. AB - A depressing synapse transforms a time interval into a voltage amplitude. The effect of that transformation on the output of the neuron and network depends on the kinetics of synaptic depression and properties of the postsynaptic neuron and network. Using as examples neural circuits that incorporate depressing synapses, we show how short-term depression can contribute to a surprising variety of time dependent computational and behavioral tasks. PMID- 15888601 TI - Neurologic manifestations in welders with pallidal MRI T1 hyperintensity. AB - BACKGROUND: Neurologic symptoms have been attributed to manganese fumes generated during welding. Increased T1 MRI signal in the basal ganglia is a biologic marker of manganese accumulation. Recent studies have associated welding and parkinsonism, but generally without MRI corroboration. OBJECTIVE: To characterize the clinical and neuropsychological features of patients with MRI basal ganglia T1 hyperintensity, who were ultimately diagnosed with neurotoxicity from welding fumes. METHODS: The medical records of welders referred to the Department of Neurology with neurologic problems and basal ganglia T1 hyperintensity were reviewed. RESULTS: All eight patients were male career welders with increased T1 basal ganglia signal on MRI of the brain. Several different clinical syndromes were recognized: a parkinsonian syndrome (three patients), a syndrome of multifocal myoclonus and limited cognitive impairment (two patients), a mixed syndrome with vestibular-auditory dysfunction (two patients), and minor subjective cognitive impairment, anxiety, and sleep apnea (one patient). Neuropsychometric testing suggested subcortical or frontal involvement. Inadequate ventilation or lack of personal respiratory protection during welding was a common theme. CONCLUSIONS: Welding without proper protection was associated with syndromes of parkinsonism, multifocal myoclonus, mild cognitive impairment, and vestibular-auditory dysfunction. The MRI T1 hyperintensity in the basal ganglia suggests that these may have been caused by manganese neurotoxicity. PMID- 15888603 TI - Appearance and disappearance of neutralizing antibodies during interferon-beta therapy. AB - BACKGROUND: Neutralizing antibodies (NABs) occur frequently in patients receiving interferon (IFN)-beta for multiple sclerosis (MS), but it is unclear whether occurrence of NABs is predictive for the persistence of NABs during continued IFN beta therapy. METHODS: The authors used an antiviral neutralization bioassay to measure NABs blindly from 6 months up to 78 months in patients with MS who were followed for at least 24 months during treatment with IFN-beta. Patients were classified into three groups: 1) persistently NAB-negative patients, defined as patients without any positive samples at any time; 2) definitely NAB-positive patients, defined as patients who had at least two consecutive positive samples; and 3) patients with fluctuating NAB-positive and NAB-negative samples. RESULTS: A total of 455 patients were included in the study. Overall, 52.3% of the patients were persistently NAB-negative, 40.9% became definitely NAB-positive, and the remaining 6.8% were fluctuating. More patients treated with IFN-beta-1a (Avonex) remained NAB-negative (p < 0.0001), whereas there was no difference between IFN-beta-1b (Betaferon) and IFN-beta-1a (Rebif). Patients who have remained NAB-negative during the first 24 months of therapy rarely developed NABs. On the contrary, the majority of patients, who had been NAB-positive from 12 through 30 months after start of therapy, remained NAB-positive. CONCLUSIONS: NABs should be measured in all patients treated with IFN-beta. If patients have been persistently NAB-negative for 24 months, measurements can be discontinued. Patients who have been NAB-positive for a period of 18 months or more usually remain NAB-positive for a long time. PMID- 15888605 TI - Oscillatory EEG correlates of episodic trace decay. AB - Recent studies suggest that human theta oscillations appear to be functionally associated with memory processes. It is less clear, however, to what type of memory sub-processes theta is related. Using a continuous word recognition task with different repetition lags, we investigate whether theta reflects the strength of an episodic memory trace or general processing demands, such as task difficulty. The results favor the episodic trace decay hypothesis and show that during the access of an episodic trace in a time window of approximately 200-400 ms, theta power decreases with increasing lag (between the first and second presentation of an item). LORETA source localization of this early theta lag effect indicates that parietal regions are involved in episodic trace processing, whereas right frontal regions may guide the process of retrieval. We conclude that episodic encoding can be characterized by two different stages: traces are first processed at parietal sites at approximately 300 ms, then further processing takes place in regions of the medial temporal lobe at approximately 500 ms. Only the first stage is related to theta, whereas the second is reflected by a slow wave with a frequency of approximately 2.5 Hz. PMID- 15888606 TI - The human parietal operculum. II. Stereotaxic maps and correlation with functional imaging results. AB - In this study we describe the localization of the cytoarchitectonic subdivisions of the human parietal operculum in stereotaxic space and relate these anatomically defined cortical areas to the location of the functionally defined secondary somatosensory cortex (SII cortex) using a meta-analysis of functional imaging results. The human parietal operculum consists of four distinct cytoarchitectonic areas (OP 1-4) as shown in the preceding publication. The 10 cytoarchitectonically examined brains were 3-D-reconstructed and spatially normalized to the T1-weighted single-subject template of the Montreal Neurological Institute (MNI). A probabilistic map was calculated for each area in this standard stereotaxic space. A cytoarchitectonic summary map of the four cortical areas on the human parietal operculum which combines these probabilistic maps was subsequently computed for the comparison with a meta-analysis of functional locations of SII. The meta-analysis used the results from 57 fMRI and PET studies and allowed the comparison of the functionally defined SII region to the cytoarchitectonic map of the parietal operculum. The functional localization of SII showed a good match to the cytoarchitectonically defined region. Therefore the cytoarchitectonic maps of OP 1-4 of the human parietal operculum can be interpreted as an anatomical correlate of the (functionally defined) human SII region. Our results also suggest that the SII foci reported in functional imaging studies may actually reflect activations in either of its architectonic subregions. PMID- 15888607 TI - The human parietal operculum. I. Cytoarchitectonic mapping of subdivisions. AB - The human secondary somatosensory cortex (SII) is located on the parietal operculum, as shown by intraoperative stimulation and functional imaging studies. The position and extent of the anatomical correlates of this functionally defined region, however, are still unknown. We have therefore histologically mapped the putative anatomical correlates of the SII cortex in cell-body-stained histological sections of 10 human postmortem brains using quantitative cytoarchitectonic analysis. The gray level index (GLI), which is an indicator of the volume fraction of nerve cell bodies, was measured in the parietal operculum. GLI profiles as measures of the laminar pattern of the cortex were extracted perpendicular to cortical layers. Cytoarchitectonic borders were detected observer-independently by multivariate statistical analysis of the laminar profiles. Four cytoarchitectonic areas (termed OP 1-4) were identified. This cytoarchitectonic heterogeneity of the parietal operculum corresponds to results of functional imaging studies on the human SII cortex and data from non-human primates where multiple subregions within SII have been demonstrated by electrophysiological and connectivity studies. PMID- 15888608 TI - Spinal changes in patients with spondyloarthritis: comparison of MR imaging and radiographic appearances. AB - Since the advent of highly effective TNF-alpha inhibitors for treating spondyloarthritides, referring rheumatologists have been requesting the sensitive visualization of inflammatory changes not only of the sacroiliac joints but of the entire spine. Given that changes in spondyloarthritis may be very subtle, their visualization by means of magnetic resonance (MR) imaging relies critically on selecting the proper imaging protocol. Spinal changes associated with spondyloarthritis are florid anterior spondylitis (or Romanus lesion), florid diskitis (or Andersson lesion), ankylosis, insufficiency fractures of the ankylosed spine, syndesmophytes, arthritis of the apophyseal and costovertebral joints, and enthesitis of the interspinal ligaments. A comparison of MR imaging findings with those of conventional radiography in individual patients reveals strengths and weaknesses of both modalities. Results of this comparison suggest that syndesmophytes are depicted better with radiography; ankylosis, equally well with both imaging techniques; and all other lesions, better with MR imaging. Classification of the different findings based on the typical signal-intensity changes seen on MR images enables standardized reporting, and scoring the lesions may be helpful in clinical trials. PMID- 15888610 TI - Hamstring muscle complex: an imaging review. AB - Increasing activity in the general population and the high demands placed on athletes have resulted in injuries to the hamstring muscle complex (HMC) being commonplace in sports. Imaging of HMC injuries can form a considerable part of a sports medicine practice, with a wide spectrum of such injuries being reflected in their varied imaging appearances. Magnetic resonance (MR) imaging and ultrasonography (US) are the imaging modalities of choice in this setting. Both MR imaging and US provide exquisitely detailed information about the HMC with respect to localization and characterization of injury. Optimization of MR imaging involves the use of a surface coil and high-resolution techniques, allowing the musculoskeletal radiologist not only to diagnose injury and assess severity but also to provide the clinician with useful clues with respect to prognosis. The portability and availability of US make it an attractive modality for the diagnosis of acute hamstring injuries, although its effectiveness is dependent on operator experience. A thorough knowledge of the HMC anatomy and of the spectrum of imaging findings in HMC injury is crucial for providing optimal patient care and will enable the musculoskeletal radiologist to make an accurate and useful contribution to the treatment of athletes at all levels of participation. PMID- 15888611 TI - Normal variants and diseases of the peroneal tendons and superior peroneal retinaculum: MR imaging features. AB - Diseases of the peroneal tendons and superior peroneal retinaculum (SPR) are frequently underdiagnosed causes of lateral ankle pain and instability. Common diseases include tenosynovitis, rupture, and dislocation of the peroneal tendons as well as injuries to the SPR. Many anatomic variants, such as a flat or convex retromalleolar fibular groove, hypertrophy of the peroneal tubercle, an accessory peroneus quartus muscle, a low-lying peroneus brevis muscle belly, or an os peroneum, may be associated with or predispose to lateral ankle disease. Magnetic resonance (MR) imaging is excellent for detecting soft-tissue and bone variants and abnormalities related to the lateral ankle. Knowledge of the MR imaging appearances of these entities aids radiologists in making the precise diagnosis of disorders of the peroneal tendons and SPR. Pitfalls and normal variants of the peroneal tendons, including magic angle phenomenon, pseudosubluxation of the peroneus brevis tendon, a bifurcated or mildly crescentic peroneus brevis tendon, insertion of the peroneus quartus tendon into the peroneus brevis tendon, and the presence of an os peroneum are important to recognize. It is also useful to be familiar with the MR imaging appearances of SPR injuries, which can be an overlooked but treatable cause of lateral ankle pain and instability. PMID- 15888612 TI - Multidetector CT of musculoskeletal disease in the pediatric patient: principles, techniques, and clinical applications. AB - Computed tomography (CT) plays an important role in the evaluation of musculoskeletal disease in the pediatric patient. With the advent of high performance 16-section multidetector CT, images can be produced with subsecond gantry rotation times and with submillimeter acquisition, which yields true isotropic high-resolution volume data sets; these features are not attainable with older spiral CT technology. Such capabilities are particularly helpful in the evaluation of pediatric patients by virtually eliminating the need for sedation and minimizing dependence on patient cooperation. The role of three dimensional (3D) volume imaging in the evaluation of pediatric musculoskeletal disease continues to evolve, with this technique becoming increasingly important in detection and characterization of lesions as well as in decisions about patient care. Specific designs and protocols for multidetector CT studies can be selected to minimize radiation dose to the patient. Principal clinical applications of 3D CT in evaluation of the pediatric musculoskeletal system include developmental abnormalities, trauma, neoplasms, and postoperative imaging. PMID- 15888613 TI - Childhood X-linked adrenoleukodystrophy: clinical-pathologic overview and MR imaging manifestations at initial evaluation and follow-up. AB - X-linked adrenoleukodystrophy (ALD) is a rare metabolic disorder caused by peroxisomal enzyme failure. Several phenotypes can be distinguished on the basis of clinical onset and manifestations. Childhood cerebral X-linked ALD is the most severe phenotype, resulting in rapid neurologic deterioration and early death. Patients with this disease may be hospitalized with far-advanced central nervous system (CNS) lesions or may complain of symptoms similar to those of certain psychiatric disorders, possibly leading to a wrong diagnosis. Although the general prognosis for patients with childhood cerebral X-linked ALD is still poor, new treatment modalities have been introduced, some of which are helpful in relieving clinical symptoms and prolonging life. With the introduction of these new therapies and increased clinical detection of childhood cerebral X-linked ALD, brain magnetic resonance (MR) imaging has become an essential tool for initial and follow-up evaluation. MR imaging allows early detection of CNS lesions and helps differentiate childhood cerebral X-linked ALD from other disorders. The characteristic MR imaging features of childhood cerebral X-linked ALD have been well documented, although most radiologists have limited experience with serial follow-up MR imaging in this context. Familiarity with the clinical pathologic manifestations and progressive MR imaging features of childhood cerebral X-linked ALD will be helpful in evaluating affected patients. PMID- 15888614 TI - Intraabdominal fetal echogenic masses: a practical guide to diagnosis and management. AB - Intraabdominal calcifications and other echogenic masses are relatively common findings during fetal sonography. Many are associated with no additional risk for the fetus or neonate. They may arise from the liver, gallbladder, spleen, kidneys, adrenal glands, gastrointestinal tract, or peritoneal cavity. Detection of such lesions should prompt a detailed survey for additional findings and a review of the maternal history. In some cases, fetal karyotyping may be indicated. In most cases, the diagnosis, management, and outcome are determined according to a combination of specific ultrasound appearances and at least one additional maternal or fetal factor. In utero diagnosis can often be achieved with careful evaluation of the lesion echotexture, associated calcifications, additional findings, and evolution over time. In most cases, expectant management is sufficient, but some patients require transfer to a facility where early postnatal intervention is available. A systematic approach to the findings aids in differential diagnosis and management. PMID- 15888615 TI - Fetal schizencephaly: pre- and postnatal imaging with a review of the clinical manifestations. AB - Schizencephaly is a rare malformation of the central nervous system. Prenatal ultrasound (US) allows diagnosis of schizencephaly, although prenatal magnetic resonance (MR) imaging is even more specific in detection of gray matter lining the defect, communication with the ventricle, and other associated structural abnormalities. Six cases of schizencephaly were evaluated at one institution; prenatal US was performed in all cases, and fetal MR imaging was performed in three cases. As with many malformations, more severe cases of schizencephaly often manifest in utero. All of the cases studied were of the open-lip variety; three cases were bilateral, and three were unilateral. The cleft was not appreciated at initial US in only one case, which consisted of a small unilateral defect, with the diagnosis made at subsequent MR imaging. A survey of the clinical literature on schizencephaly shows that the severity of the motor and mental impairments is directly related to the extent of the anatomic defect. The differential diagnosis for a cerebrospinal fluid-containing abnormality of the fetal brain includes both developmental and destructive lesions. Prenatal detection of schizencephaly can assist in management of the pregnancy. PMID- 15888616 TI - Fibropolycystic liver disease: CT and MR imaging findings. AB - Fibropolycystic liver disease encompasses a spectrum of related lesions of the liver and biliary tract that are caused by abnormal embryologic development of the ductal plates. These lesions (congenital hepatic fibrosis, biliary hamartomas, autosomal dominant polycystic disease, Caroli disease, choledochal cysts) can be clinically silent or can cause signs and symptoms such as cholangitis, portal hypertension, gastrointestinal bleeding, infections, and space-occupying masses. The different types of fibropolycystic liver disease demonstrate characteristic findings at computed tomography (CT) and magnetic resonance (MR) imaging. Patients with congenital hepatic fibrosis typically have imaging evidence of liver morphologic abnormalities, varices, splenomegaly, renal lesions, and other associated ductal plate abnormalities. Biliary hamartomas usually manifest as multiple cysts that are nearly uniform in size and measure up to 15 mm in diameter. Autosomal dominant polycystic disease typically manifests as an enlarged and diffusely cystic liver. In Caroli disease, cystic or fusiform dilatation of the intrahepatic ducts is seen, as well as the "central dot sign," which corresponds to a portal vein branch protruding into the lumen of a dilated bile duct. Choledochal cyst manifests as a fusiform or cystic dilatation of the extrahepatic bile duct. Awareness of these CT and MR imaging features is essential in detecting and differentiating between various fibropolycystic liver diseases and can assist in proper management. PMID- 15888617 TI - Imaging diagnosis of cystic pancreatic lesions: pseudocyst versus nonpseudocyst. AB - Although the clinical, radiologic, and pathologic features of cystic pancreatic lesions are well known, preoperative diagnosis is difficult. Differentiation between a pancreatic pseudocyst and a cystic pancreatic neoplasm is crucial in determining the proper treatment. Careful evaluation of the patient's clinical history is important for accurate diagnosis of a pseudocyst. Clinical scenarios include a pseudocyst developing after acute pancreatitis and a pseudocyst superimposed on chronic pancreatitis. However, a pseudocyst in a patient with no clinical history of pancreatitis poses a diagnostic problem. The differential diagnosis of a neoplastic cystic lesion of the pancreas includes serous cystadenoma, mucinous cystic neoplasms, intraductal papillary mucinous tumor, and solid and papillary epithelial neoplasm. Definitive diagnosis is often possible when the lesion has a typical radiologic appearance, but in many cases characterization with imaging alone is impossible. Thin-section computed tomography with multiplanar reformation, magnetic resonance cholangiopancreatography, and endoscopic ultrasonography have emerged as modalities that can provide additional diagnostic information. Familiarity with the range of imaging appearances and awareness of the diagnostic strengths and limitations of each imaging modality are important for accurate diagnosis and management of cystic pancreatic lesions. PMID- 15888618 TI - Contrast-enhanced MR angiography for evaluation of vascular complications of the pancreatic transplant. AB - Vascular complications are a common cause of postoperative dysfunction in a pancreatic transplant. Coronal three-dimensional (3D) contrast material-enhanced magnetic resonance (MR) angiography performed with high spatial and temporal resolution is a safe and effective method of assessing these vascular complications. A study was performed of selected patients who had undergone MR imaging and MR angiography during the past 6 years for evaluation of graft dysfunction following pancreatic transplantation. Thrombosis within peripheral stump vessels involving either the arterial or venous segments was a commonly observed vascular complication. Isolated distal arterial stump thrombi are incidental findings that may not require treatment, whereas venous stump thrombi may become clinically significant in patients in whom clot propagates proximally to occlude draining pancreatic veins and are typically treated with anticoagulants or thrombectomy. Because it is difficult to predict which patients will experience clot propagation, patients with venous stump thrombi may be followed up with serial imaging regardless of treatment initiated at presentation. Although susceptibility artifacts can mimic anastomotic stenoses at MR imaging, careful attention to the multiple sequences used allows recognition of this potential pitfall. Contrast-enhanced 3D MR angiography is an accurate method of evaluating the vascular anatomy of pancreatic transplants and can help guide clinical management. PMID- 15888619 TI - Imaging of small bowel disease: comparison of capsule endoscopy, standard endoscopy, barium examination, and CT. AB - Capsule endoscopy is a revolutionary new diagnostic tool for the detection of small bowel disease. As the name implies, capsule endoscopy makes use of a swallowable video capsule; as such, it is the only technique that allows noninvasive endoscopic examination of the entire small bowel without sedation. Obscure gastrointestinal bleeding is the most common indication for capsule endoscopy, which commonly depicts arteriovenous malformations, small bowel tumors, and ulcers missed at standard endoscopy and imaging examinations. However, capsule endoscopy is not optimal for the localization of small bowel lesions. In addition, lesions can be missed due to poor bowel preparation, rapid or delayed small bowel transit, or orientation of the camera away from a lesion. Computed tomography and barium examinations are useful for detecting these missed lesions and for localizing lesions detected at capsule endoscopy. Other limitations of capsule endoscopy are the inability to treat lesions and its limited use in patients with small bowel strictures or obstruction. Nevertheless, this new technique is easy to perform, is well tolerated by patients, and, for the first time, allows noninvasive endoscopic evaluation of the entire small bowel. PMID- 15888621 TI - Unusual nonneoplastic peritoneal and subperitoneal conditions: CT findings. AB - Peritoneal disease can manifest at computed tomography (CT) as fluid accumulation within the peritoneal cavity (ascites) or soft-tissue infiltration of the various peritoneal ligaments and mesenteries. Beyond the commonly encountered cases of typical ascites and peritonitis, there is a wide spectrum of uncommon nonneoplastic conditions that may involve the peritoneal and subperitoneal spaces. For example, systemic or organ-based diseases that occasionally involve the peritoneum include eosinophilic gastroenteritis, amyloidosis, extramedullary hematopoiesis, Erdheim-Chester disease, sarcoidosis, and mesenteric cavitary lymph node syndrome. Tumorlike conditions that may affect the peritoneum include aggressive fibromatosis (desmoid), inflammatory pseudotumor, retractile mesenteritis, and Castleman disease. Atypical peritoneal infections include tuberculosis, actinomycosis, echinococcosis, Whipple disease, and mesenteric adenitis. Conditions involving the subperitoneal fat include epiploic appendagitis, mesenteric panniculitis, and segmental omental infarction, all of which have characteristic CT findings. CT is an excellent imaging modality for detection and characterization of peritoneal involvement from these unusual diseases. PMID- 15888622 TI - Color Doppler US of the postoperative penis: anatomy and surgical complications. AB - A number of surgical procedures that significantly change the penile anatomy and vasculature can be used to manage pathologic conditions of the penis (eg, congenital and acquired deformities, erectile dysfunction, priapism). Phallic reconstruction surgery can be used for sex reassignment and after penile amputation or for correction of congenital malformations. Color Doppler ultrasonography (US) clearly depicts the normal penile anatomy and postoperative changes (eg, changes of the tunica albuginea, extraalbugineal pathologic fluid collections, cavernosal tissue changes produced by scars and fibrosis). It is also effective in evaluating surgery-related complications and determining the causes of erectile dysfunction and other unsatisfactory long-term results. Moreover, color Doppler US of the penile vessels and vascular anastomoses following revascularization allows direct evaluation of flow characteristics, shunt patency, and venous engorgement. Color Doppler US is the imaging modality of choice in evaluating patients who have undergone penile surgery. PMID- 15888623 TI - Cross-sectional imaging of the female urethra: technique and results. AB - Clinical assessment of women with urethral symptoms is difficult, necessitating further evaluation with imaging. Urethrography provides limited information on luminal abnormalities of the urethra. Recent advances in ultrasound (US) and magnetic resonance (MR) imaging have dramatically improved evaluation of the female urethra, clarifying findings at physical examination and providing accurate road maps for surgeons. High-resolution transvaginal US, transperineal US, and transurethral US are reliable techniques for diagnosis and characterization of urethral abnormalities. High-resolution multiplanar MR imaging with phased-array pelvic and endovaginal coils demonstrates the urethral anatomy in greater detail. In women with urethral diverticula, US and MR imaging demonstrate the number of diverticula and the location, size, configuration, and possible contents of the sac. Most important, the position of the neck of the diverticulum may be identified for the surgeon. Imaging features do not allow differentiation between histologic subtypes of urethral carcinoma; the diagnosis is established with histopathologic examination. Periurethral cysts do not communicate with the urethra and therefore can often be differentiated from urethral diverticula at endocavitary MR imaging. High-resolution multiplanar US and MR imaging allow comprehensive evaluation of abnormalities of the female urethra. PMID- 15888624 TI - Imaging-guided percutaneous biopsy of mediastinal lesions: different approaches and anatomic considerations. AB - Percutaneous needle biopsy with imaging guidance allows access to lesions in virtually all mediastinal locations. A direct mediastinal approach, which enables extrapleural needle placement, is the preferred method to avoid the risk of pneumothorax. Techniques that allow extrapleural access include the parasternal, paravertebral, transsternal, and suprasternal approaches, which are performed with computed tomographic or ultrasonographic guidance. The parasternal approach is used for biopsy of anterior or middle mediastinal lesions when the lesion or intervening mediastinal fat extends to the anterior chest wall, lateral to the sternum; injury to the internal mammary vessels is a potential complication. The paravertebral approach is used for biopsy of subcarinal and other posterior mediastinal lesions; saline solution is often injected to widen the mediastinum. The transsternal approach, which involves needle placement through the sternum, is used for biopsy of anterior or middle mediastinal lesions that are not accessible with the parasternal approach. Biopsy of superior mediastinal lesions can be performed with a suprasternal approach. An alternative to these direct mediastinal approaches involves advancing the needle through a pleural space created by an existing pleural effusion or iatrogenic pneumothorax. Another alternative is the transpulmonary approach, which involves transgression of the lung and visceral pleura by the needle and is associated with a substantial risk of pneumothorax. PMID- 15888626 TI - Tree-in-bud pattern at thin-section CT of the lungs: radiologic-pathologic overview. AB - The tree-in-bud pattern is commonly seen at thin-section computed tomography (CT) of the lungs. It consists of small centrilobular nodules of soft-tissue attenuation connected to multiple branching linear structures of similar caliber that originate from a single stalk. Originally reported in cases of endobronchial spread of Mycobacterium tuberculosis, this pattern is now recognized as a CT manifestation of many diverse entities. These entities include peripheral airway diseases such as infection (bacterial, fungal, viral, or parasitic), congenital disorders, idiopathic disorders (obliterative bronchiolitis, panbronchiolitis), aspiration or inhalation of foreign substances, immunologic disorders, and connective tissue disorders and peripheral pulmonary vascular diseases such as neoplastic pulmonary emboli. Knowledge of the many causes of this pattern can be useful in preventing diagnostic errors. In addition, although the causes of this pattern are frequently indistinguishable at radiologic evaluation, the presence of additional radiologic findings, along with the history and clinical presentation, can often be useful in suggesting the appropriate diagnosis. PMID- 15888627 TI - From the archives of the AFIP: lymphangioleiomyomatosis: radiologic-pathologic correlation. AB - Lymphangioleiomyomatosis (LAM) is an uncommon interstitial lung disease that exclusively affects women, usually during their reproductive years. LAM is characterized pathologically by abnormal proliferation of LAM cells in the lungs and in thoracic and retroperitoneal lymphatics. Thirty-three cases of LAM were reviewed retrospectively for clinical and radiologic findings. Twenty-eight (85%) of 33 women (aged 21-62 years; mean, 37.5 years) were symptomatic. Radiographs (n = 32) demonstrated reticular opacities in 21 (66%) patients, large lung volumes in 17 (53%), pleural effusion in 14 (44%), and pneumothorax in 13 (41%). High resolution CT (n = 15) and conventional CT (n = 3) showed 2-5-mm bilateral thin walled cysts in all patients and cysts that were 6-12 mm or larger in patients with severe lung involvement. CT depicted diffuse lung involvement by cysts in nine (50%) patients, relative sparing of lung apices in seven (39%), and relative sparing of lung bases in two (11%). Pleural effusion and pneumothorax were seen at CT in four (22%) and three (17%) patients, respectively. Four cases of tuberous sclerosis complex-associated LAM (TSC-LAM) (women aged 27-50 years; mean, 35.7 years) were similarly reviewed. Three (75%) were symptomatic. Radiographs (n = 4) demonstrated reticular opacities in three (75%) and large lung volumes in two (50%). All high-resolution CT (n = 3) and conventional CT (n = 1) studies showed 2-5-mm bilateral thin-walled cysts and cysts that were 6-12 mm or larger in two patients with severe lung involvement. Pleural effusion and pneumothorax were demonstrated at CT in three (75%) and two (50%) patients, respectively. LAM and TSC-LAM affect symptomatic women who often exhibit reticular opacities and large lung volumes at radiography and bilateral uniform small thin-walled cysts at CT. Large (>12 mm) cysts occur in patients with severe cystic lung involvement. Pneumothorax and pleural effusion are common associated findings. PMID- 15888628 TI - Best cases from the AFIP: primary hyperparathyroidism due to parathyroid adenoma. PMID- 15888629 TI - Best cases from the AFIP: bilateral testicular tumors: seminoma and mixed germ cell tumor. PMID- 15888630 TI - Informatics in radiology (infoRAD): new tools for computer assistance in thoracic CT part 2. Therapy monitoring of pulmonary metastases. AB - Owing to the rapid development of scanner technology, thoracic computed tomography (CT) offers new possibilities but also faces enormous challenges with respect to the quality of computer-assisted diagnosis and therapy planning. In the framework of the Virtual Institute for Computer Assistance in Clinical Radiology cooperative research project, a software application was developed to assist the radiologist in the analysis of thoracic CT data for the purpose of evaluating the response to tumor therapy. The application provides follow-up support for monitoring of tumor therapy by means of volumetric quantification of tumors and temporal registration. In addition, anatomically adequate three dimensional visualization techniques for convenient examination of large data sets are included. With close cooperation between computer scientists and radiologists, the application was tested and optimized to achieve a high degree of usability. Several clinical studies were carried out, the results of which indicated that the application improves therapy monitoring with respect to accuracy and time required. PMID- 15888631 TI - Informatics in radiology (infoRAD): benefits of content-based visual data access in radiology. AB - The field of medicine is often cited as an area for which content-based visual retrieval holds considerable promise. To date, very few visual image retrieval systems have been used in clinical practice; the first applications of image retrieval systems in medicine are currently being developed to complement conventional text-based searches. An image retrieval system was developed and integrated into a radiology teaching file system, and the performance of the retrieval system was evaluated, with use of query topics that represent the teaching database well, against a standard of reference generated by a radiologist. The results of this evaluation indicate that content-based image retrieval has the potential to become an important technology for the field of radiology, not only in research, but in teaching and diagnostics as well. However, acceptance of this technology in the clinical domain will require identification and implementation of clinical applications that use content-based access mechanisms, necessitating close cooperation between medical practitioners and medical computer scientists. Nevertheless, content-based image retrieval has the potential to become an important technology for radiology practice. PMID- 15888632 TI - Gallery of medical devices: part 1: orthopedic devices for the extremities and pelvis. PMID- 15888633 TI - Identification of the hydrophobic glycoproteins of Caenorhabditis elegans. AB - Hydrophobic proteins such as integral membrane proteins are difficult to separate, and therefore to study, at a proteomics level. However, the Asn-linked (N-linked) carbohydrates (N-glycans) contained in membrane glycoproteins are important in differentiation, embryogenesis, inflammation, cancer and metastasis, and other vital cellular processes. Thus, the identification of these proteins and their sites of glycosylation in a well-characterized model organism is the first step toward understanding the mechanisms by which N-glycans and their associated proteins function in vivo. In this report, a proteomics method recently developed by our group was applied to identify 117 hydrophobic N glycosylated proteins of Caenorhabditis elegans extracts by analysis of 195 glycopeptides containing 199 Asn-linked oligosaccharides. Most of the proteins identified are involved in cell adhesion, metabolism, or the transport of small molecules. In addition, there are 18 proteins for which no function is known or predictable by sequence homologies and two proteins which were previously predicted to exist only on the basis of genomic sequences in the C. elegans database. Because N-glycosylation is initiated in the lumen of the endoplasmic reticulum (ER), our data can be used to reassess the previously predicted subcellular localizations of these proteins. As well, the identification of N glycosylation sites helps establish the membrane topology of the associated glycoproteins. Caenorhabditis elegans strains are presently available with mutations in 17 of the genes we have identified. The powerful genetic tools available for C. elegans can be used to make other strains with mutations in genes encoding N-glycosylated proteins and thereby determine N-glycan function. PMID- 15888634 TI - Banana lectin is unique in its recognition of the reducing unit of 3-O-beta glucosyl/mannosyl disaccharides: a calorimetric study. AB - The binding of banana lectin (BanLec) to laminaribiose (Glcbeta1,3Glc) and a series of novel synthetic analogues was measured by titration calorimetry to assess the contribution of the hydroxyl groups of the reducing glycosyl moiety and its 3-O-beta-substituent to binding. Key areas of interaction involved the 1, 2, and 6 positions of the reducing-terminal hexose unit. The alpha-anomeric configuration of the reducing hexose was strongly favored over the beta-anomer. The 2-hydroxyl in the axial position (mannose) also enhanced binding, whereas the 6-hydroxymethyl group was essential, because xylopyranose in the reducing position was inactive. The 3-O-beta-glucosyl unit of methyl alpha-laminaribioside could be replaced by any of its monodeoxy derivatives. However, the 4'-deoxy derivative or axial hydroxy (galactosyl) substitution was somewhat detrimental to binding. 3-O-substitution with the (S)tetrahydropyranyl ring or a benzyl group had similar effect as 4'-deoxyglucosyl substitution. Surprisingly, p-nitrobenzyl or beta-xylosyl 3-O-substitution greatly enhanced binding of the reducing glucosyl or mannosyl derivative. Chemical syntheses of a number of novel disaccharides and analogues prepared for this study are described. PMID- 15888635 TI - Community-based research partnerships: challenges and opportunities. AB - The complexity of many urban health problems often makes them ill suited to traditional research approaches and interventions. The resultant frustration, together with community calls for genuine partnership in the research process, has highlighted the importance of an alternative paradigm. Community-based participatory research (CBPR) is presented as a promising collaborative approach that combines systematic inquiry, participation, and action to address urban health problems. Following a brief review of its basic tenets and historical roots, key ways in which CBPR adds value to urban health research are introduced and illustrated. Case study examples from diverse international settings are used to illustrate some of the difficult ethical challenges that may arise in the course of CBPR partnership approaches. The concepts of partnership synergy and cultural humility, together with protocols such as Green et al.'s guidelines for appraising CBPR projects, are highlighted as useful tools for urban health researchers seeking to apply this collaborative approach and to deal effectively with the difficult ethical challenges it can present. PMID- 15888636 TI - Poverty-related stressors and HIV/AIDS transmission risks in two South African communities. AB - Community stress associated with poverty is related to health risks and poor health outcomes. Perceived community stress is specifically related to HIV transmission risk behaviors in the United States, but research has not examined these relationships in southern Africa, the region of the world with the highest rates of HIV infection and among the greatest poverty. Men (N=464) and women (N=531) living in impoverished adjacent communities distinguished by race (e.g., indigenous African and Coloured) completed anonymous surveys of perceptions of 10 poverty-related community stressors and measures of HIV risk-related behaviors. Indigenous African and Coloured communities differed in their perceptions of stressors, with Africans consistently viewing the 10 community stressors as more serious problems. In addition, perceived seriousness of lacking basic living resources was related to higher risk for HIV among Africans. Perceived community stress was also related to alcohol and drug use, but substance use did not mediate the association between perceived community stress and HIV risks. In the Coloured community, perceived community stressors were related to drug use, but perceived community stressors were not associated with HIV risks. These findings extend the findings of previous research to show that poverty-related stressors are associated with HIV transmission risks in some poverty-stricken communities and that these associations are not mediated by substance use. PMID- 15888637 TI - Mortality and socioeconomic deprivation in census tracts of an urban setting in southern Europe. AB - In southern European cities, research on deprivation and mortality inequalities using small-area analysis is recent. In many countries, the census tract (CT) is the smallest territorial unit for which population data are available. The aim of this study was to examine the association between mortality from all causes and socioeconomic deprivation in CTs in Barcelona (Spain). A cross-sectional ecologic study was carried out using mortality data for 1987-1995 and 1991 census variables. Mortality data were obtained from death certificates. Socioeconomic deprivation indicators were drawn from the census and included unemployment, inadequate education, and low social class. They were correlated, and a deprivation index was elaborated with them. The analysis was descriptive, and multivariate Poisson regression models were adjusted. The most deprived CTs tend to present higher mortality (49.7% of CT in the quartile associated with greatest deprivation were included in the top male mortality quartile and 40.4% in the top female mortality quartile), whereas the less deprived ones present lower mortality. For male mortality, the risk of dying among those in the quartile representing most deprivation is from 25 to 29% higher (depending on the indicator chosen) than the least deprived quartile, and for women, it is from 12 to 14% higher. We concluded that the mortality from all causes in the CT of a southern European city has shown a clear positive association with a variety of socioeconomic deprivation indicators drawn from the census. Studies of this nature may help to orient more specific studies in which CTs are grouped together as a function of particular population and/or health characteristics. PMID- 15888638 TI - Use of a patient navigator to increase colorectal cancer screening in an urban neighborhood health clinic. AB - Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. Racial disparities in CRC incidence and mortality have been well documented. In addition, lower rates of CRC screening among ethnic minorities have been reported. Therefore, we tested the effectiveness of a patient navigator (PN) in increasing compliance with CRC screening in a minority community health setting. Men and women aged 50 or older attending a primary care practice were enrolled if they had not had a fecal occult blood test within the past year, a sigmoidoscopy or barium enema within the past 3-5 years, or a colonoscopy within the past 10 years. Participants were randomly assigned either to receive navigator services (PN+) or not to receive navigator services (PN-). There were no demographic differences between the two groups. Within 6 months of physician recommendation, 15.8% in the PN+group had complied with an endoscopic examination, compared with only 5% in the PN - group (P=.019). The PN+group also demonstrated higher rates of fecal occult blood test completion (42.1% vs. 25%, P=.086). Thus, a PN system successfully increases CRC screening rates among a predominantly minority population of low socioeconomic status. PMID- 15888639 TI - Awareness of heart attack symptoms and lifesaving actions among New York City area residents. AB - The American Heart Association has a national network of community-based programs designed to reduce response times to cardiac emergencies by improving access to automatic external defibrillators (AEDs) among laypersons. Success of these Operation Heartbeat programs depends in part on the public's knowledge of the warning signs of a myocardial infarction (MI) and appropriate response to cardiac arrest victims. In May 2000, a 7-minute telephone survey was administered to a random sample of adults residing within the American Heart Association affiliate territories of New York, New Jersey, and Connecticut to determine the knowledge of MI symptoms, confidence in cardiopulmonary resuscitation (CPR) use, and the awareness of AEDs. Of the respondents, 60% were women (n=1,128), 83% were Caucasians (n=1,558), 15.2% were non-whites (African American, Asian, or Hispanic), and 38.5% had at least a college degree (n=724). Women were significantly more likely than men to know that sex differences exist in the warning signs for an MI (63% vs. 30.7%, respectively; P< .001). Whites had above average confidence in MI recognition compared with non-whites (39.2% vs. 27.4%, respectively; P< .001) and were more cognizant of the public availability of AEDs (54.5% vs. 33.2%, respectively; P< .001). Our findings suggest that racial/ethnic and sex disparities exist in the awareness of AEDs and in the knowledge of atypical MI symptoms in women, respectively. Innovative CPR outreach programs might be needed in New York area communities to increase CPR training among all adults, to increase AED awareness in vulnerable populations, and to improve knowledge and confidence in the recognition of acute MI symptoms. PMID- 15888640 TI - Disparities by race in heat-related mortality in four US cities: the role of air conditioning prevalence. AB - Daily mortality is typically higher on hot days in urban areas, and certain population groups experience disproportionate risk. Air conditioning (AC) has been recommended to mitigate heat-related illness and death. We examined whether AC prevalence explained differing heat-related mortality effects by race. Poisson regression was used to model daily mortality in Chicago, Detroit, Minneapolis, and Pittsburgh. Predictors included natural splines of time (to control seasonal patterns); mean daily apparent temperature on the day of death, and averaged over lags 1-3; barometric pressure; day of week; and a linear term for airborne particles. Separate, city-specific models were fit to death counts stratified by race (Black or White) to derive the percent change in mortality at 29 degrees C, relative to 15 degrees C (lag 0). Next, city-specific effects were regressed on city- and race-specific AC prevalence. Combined effect estimates across all cities were calculated using inverse variance-weighted averages. Prevalence of central AC among Black households was less than half that among White households in all four cities, and deaths among Blacks were more strongly associated with hot temperatures. Central AC prevalence explained some of the differences in heat effects by race, but room-unit AC did not. Efforts to reduce disparities in heat related mortality should consider access to AC. PMID- 15888641 TI - Public health priorities at subnational levels. PMID- 15888642 TI - Behavior problems associated with sleep disordered breathing in school-aged children--the Tucson children's assessment of sleep apnea study. AB - OBJECTIVE: The purpose of the current study was to examine prevalence of and relations between a commonly used measure of nighttime breathing problems, the Respiratory Disturbance Index (RDI), and a range of problem behaviors in community children. METHODS: Participants were 403 unreferred children aged 6-12 years. Recruitment was completed through public elementary schools. Overnight unattended in-home polysomnography was used to assess sleep and breathing. The RDI was used as the indicator of respiratory events during sleep. The Child Behavior Checklist and the Conners' Parent Rating Scales-Revised were used to assess behavior. RESULTS: Prevalence rates for Attention, Cognitive Problems, Aggression, Oppositional behavior, and Social Problems were greatest for subjects with high RDIs. Prevalence for Internalizing behaviors was not greater for those subjects with high RDIs. Hyperactivity was not strongly related to higher RDIs. CONCLUSIONS: Behavioral problems may exist in the presence of nocturnal breathing events in unreferred children. Specific patterns of behavioral morbidity have still not been established. Some behaviors, such as hyperactivity, may show differing sensitivity and specificity in relation to the RDI. PMID- 15888643 TI - Constructing a prospective model of psychosocial adaptation in young adolescents with spina bifida: an application of optimal data analysis. AB - OBJECTIVE: To examine how individual- and family-level predictors in late childhood and preadolescence relate to psychosocial adaptation (i.e., scholastic success, social acceptance, and positive self-worth) in early adolescence. METHOD: This prospective longitudinal study includes 68 families of children with spina bifida and 68 comparison families of healthy children. Multimethod, multiinformant data were evaluated via optimal data analysis (ODA) and classification tree analysis (CTA) techniques. RESULTS: Factors best predicting psychosocial adaptation in early adolescence included (a) intrinsic motivation, (b) estimated verbal IQ, (c) behavioral conduct, (d) coping style, and (e) physical appearance. There were no significant group (spina bifida vs. able bodied) effects. CONCLUSIONS: The final classification model correctly classified 77.8% of the total sample, indicating that this model had significant predictive capabilities. Results suggested that processes leading to psychosocial adaptation may be similar for youth with and without chronic illness. PMID- 15888644 TI - Cortical ensemble adaptation to represent velocity of an artificial actuator controlled by a brain-machine interface. AB - Monkeys can learn to directly control the movements of an artificial actuator by using a brain-machine interface (BMI) driven by the activity of a sample of cortical neurons. Eventually, they can do so without moving their limbs. Neuronal adaptations underlying the transition from control of the limb to control of the actuator are poorly understood. Here, we show that rapid modifications in neuronal representation of velocity of the hand and actuator occur in multiple cortical areas during the operation of a BMI. Initially, monkeys controlled the actuator by moving a hand-held pole. During this period, the BMI was trained to predict the actuator velocity. As the monkeys started using their cortical activity to control the actuator, the activity of individual neurons and neuronal populations became less representative of the animal's hand movements while representing the movements of the actuator. As a result of this adaptation, the animals could eventually stop moving their hands yet continue to control the actuator. These results show that, during BMI control, cortical ensembles represent behaviorally significant motor parameters, even if these are not associated with movements of the animal's own limb. PMID- 15888645 TI - Human embryonic stem cell-derived oligodendrocyte progenitor cell transplants remyelinate and restore locomotion after spinal cord injury. AB - Demyelination contributes to loss of function after spinal cord injury, and thus a potential therapeutic strategy involves replacing myelin-forming cells. Here, we show that transplantation of human embryonic stem cell (hESC)-derived oligodendrocyte progenitor cells (OPCs) into adult rat spinal cord injuries enhances remyelination and promotes improvement of motor function. OPCs were injected 7 d or 10 months after injury. In both cases, transplanted cells survived, redistributed over short distances, and differentiated into oligodendrocytes. Animals that received OPCs 7 d after injury exhibited enhanced remyelination and substantially improved locomotor ability. In contrast, when OPCs were transplanted 10 months after injury, there was no enhanced remyelination or locomotor recovery. These studies document the feasibility of predifferentiating hESCs into functional OPCs and demonstrate their therapeutic potential at early time points after spinal cord injury. PMID- 15888646 TI - The neurosteroid allopregnanolone promotes proliferation of rodent and human neural progenitor cells and regulates cell-cycle gene and protein expression. AB - Our previous research demonstrated that the neuroactive progesterone metabolite allopregnanolone (3alpha-hydroxy-5alpha-pregnan-20-one) rapidly induced hippocampal neuron neurite regression (Brinton, 1994). We hypothesized that allopregnanolone-induced neurite regression was a prelude to mitogenesis initiated by a rise in intracellular calcium. Supporting this hypothesis, the current data demonstrate that allopregnanolone, in a dose-dependent manner, induces a significant increase in proliferation of neuroprogenitor cells (NPCs) derived from the rat hippocampus and human neural stem cells (hNSCs) derived from the cerebral cortex. Proliferation was determined by incorporation of bromodeoxyuridine and [3H]thymidine, fluorescence-activated cell sorter analysis of murine leukemia virus-green fluorescent protein-labeled mitotic NPCs, and total cell number counting. Allopregnanolone-induced proliferation was isomer and steroid specific, in that the stereoisomer 3beta-hydroxy-5beta-pregnan-20-one and related steroids did not increase [3H]thymidine uptake. Immunofluorescent analyses for the NPC markers nestin and Tuj1 indicated that newly formed cells were of neuronal lineage. Furthermore, microarray analysis of cell-cycle genes and real-time reverse transcription-PCR and Western blot validation revealed that allopregnanolone increased the expression of genes that promote mitosis and inhibited the expression of genes that repress cell proliferation. Allopregnanolone-induced proliferation was antagonized by the voltage-gated L type calcium channel (VGLCC) blocker nifedipine, consistent with the finding that allopregnanolone induces a rapid increase in intracellular calcium in hippocampal neurons via a GABA type A receptor-activated VGLCC (Son et al., 2002). These data demonstrate that allopregnanolone significantly increased rat NPC and hNSC proliferation with concomitant regulation in mitotic cell-cycle genes via a VGLCC mechanism. The therapeutic potential of allopregnanolone as a neurogenic molecule is discussed. PMID- 15888647 TI - Mouse Period1 (mPER1) acts as a circadian adaptor to entrain the oscillator to environmental light/dark cycles by regulating mPER2 protein. AB - Mouse period1 (mPer1) and mPer2 are mammalian homologs of the Drosophila clock gene period that show robust oscillation in the suprachiasmatic nucleus, the mammalian master clock, and have been implicated as essential components of the core clock mechanism. Gene-targeting studies have demonstrated that mPer2 plays a dominant function in behavioral rhythm generation, although the role of mPer1 has not been fully clarified. Here, we report that prolongation of the lighting period (4-16 h) induces a larger-delay phase shift of the behavioral rhythm in mPer1-deficient (mPer1-/-) mice. During the light-elongation task, mPER2 protein decay in mPer1-/- mice is slower (approximately 4 h) than in wild-type mice, which thereby causes larger behavioral phase delay. mPer1-/- mice could not adapt to environmental light/dark cycles in long complete photoperiods with dim light or in long skeleton photoperiods. These photoperiodic conditions mimic natural environmental changes present at high latitudes, indicating that mPer1 could operate in the adaptation of the circadian clock of nocturnal mice to large seasonal changes of environmental light/dark cycles. PMID- 15888648 TI - Pedunculopontine tegmental nucleus controls conditioned responses of midbrain dopamine neurons in behaving rats. AB - Midbrain dopamine (DA) neurons respond to sensory cues that predict reward. We tested the hypothesis that projections from the pedunculopontine tegmental nucleus (PPTg) are involved in driving this DA cell activity. First, the activity of PPTg and DA neurons was compared in a cued-reward associative learning paradigm. The majority of PPTg neurons showed phasic responses to the onset of sensory cues, at significantly shorter latency than DA cells, consistent with a PPTg-to-DA transmission of information. However, unlike DA cells, PPTg responses were almost entirely independent of whether signals were associated with rewards. Second, DA neuron responses to the cues were recorded in free-moving rats during reversible inactivation of the PPTg by microinfusion of local anesthetic. The results showed clear suppression of conditioned sensory responses of DA neurons after PPTg inactivation that was not seen after saline infusion or in non-DA cells. We propose that the PPTg relays information about the precise timing of attended sensory events, which is integrated with information about reward context by DA neurons. PMID- 15888649 TI - Central suppression of regenerated proprioceptive afferents. AB - Long after a cut peripheral nerve reinnervates muscle and restores force production in adult cats, the muscle does not respond reflexively to stretch. Motivated by the likelihood that stretch areflexia is related to problems with sensing and controlling limb position after peripheral neuropathies, we sought to determine the underlying mechanism. Electrophysiological and morphological measurements were made in anesthetized rats having one of the nerves to the triceps surae muscles either untreated or cut and immediately rejoined surgically many months earlier. First, it was established that reinnervated muscles failed to generate stretch reflexes, extending observations of areflexia to a second species. Next, multiple elements in the sensorimotor circuit of the stretch reflex were examined in both the PNS and CNS. Encoding of muscle stretch by regenerated proprioceptive afferents was remarkably similar to normal, although we observed some expected abnormalities, e.g., increased length threshold. However, the robust stretch-evoked sensory response that arrived concurrently at the CNS in multiple proprioceptive afferents produced synaptic responses that were either smaller than normal or undetectable. Muscle stretch failed to evoke detectable synaptic responses in 13 of 22 motoneurons, although electrical stimulation generated monosynaptic excitatory postsynaptic potentials that were indistinguishable from normal. The ineffectiveness of muscle stretch was not attributable therefore to dysfunction at synapses made between regenerated Ia afferents and motoneurons. Among multiple candidate mechanisms, we suggest that centrally controlled neural circuits may actively suppress the sensory information encoded by regenerated proprioceptive afferents to prevent recovery of the stretch reflex. PMID- 15888650 TI - Integration time in a subset of spinal lamina I neurons is lengthened by sodium and calcium currents acting synergistically to prolong subthreshold depolarization. AB - Lamina I of the spinal dorsal horn plays an important role in processing and relaying nociceptive information to the brain. It comprises physiologically distinct cell types that process information in fundamentally different ways: tonic neurons fire repetitively during stimulation and display prolonged EPSPs, suggesting operation as integrators, whereas single-spike neurons act like coincidence detectors. Using whole-cell recordings from a rat spinal slice preparation, we set out to determine the basis for prolonged EPSPs in tonic cells and the implications for signal processing. Kinetics of synaptic currents could not explain differences in EPSP kinetics. Instead, tonic neurons were found to express a persistent sodium current, I(Na,P), that amplified and prolonged depolarization in response to brief stimulation. Tonic neurons also expressed a persistent calcium current, I(Ca,P), that contributed to prolongation but not to amplification. Simulations using NEURON software demonstrated that I(Na,P) was necessary and sufficient to explain amplification, whereas I(Na,P) and I(Ca,P) acted synergistically to prolong depolarization: initial activation of the slower current (I(Ca,P)) depended on the faster current (I(Na,P)) but maintained activation of the faster current likewise depended on the slower current. Additional investigation revealed that I(Na,P) and I(Ca,P) could dramatically increase integration time (>30x) and thereby encourage temporal summation but at the expense of spike time precision. Thus, by prolonging subthreshold depolarization, intrinsic inward currents allow tonic neurons in spinal lamina I to specialize as integrators that are optimally suited to encode stimulus intensity. PMID- 15888651 TI - A noncanonical release of GABA and glutamate modulates neuronal migration. AB - Immature neurons express GABA and glutamate receptors before synapse formation, and both transmitters are released at an early developmental stage. We have now tested the hypothesis that the ongoing release of GABA and glutamate modulates neuronal migration. Using 5-bromo-2'-deoxyuridine labeling and cocultures of hippocampal slices obtained from naive and green fluorescent protein-transgenic mice, we report that migration is severely affected by GABA(A) or NMDA receptor antagonist treatments. These effects were also present in munc18-1 knock-out slices in which soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-dependent vesicular secretion of transmitters has been deleted. GABA(A) antagonists were more efficient than NMDA antagonists to reduce cell migration, in keeping with the earlier maturation of GABAergic mechanisms. We conclude that GABA and, to a lesser degree, glutamate released in a SNARE independent mechanism exert a paracrine action on neuronal migration. PMID- 15888652 TI - In vivo performance of genetically encoded indicators of neural activity in flies. AB - Genetically encoded fluorescent probes of neural activity represent new promising tools for systems neuroscience. Here, we present a comparative in vivo analysis of 10 different genetically encoded calcium indicators, as well as the pH sensitive synapto-pHluorin. We analyzed their fluorescence changes in presynaptic boutons of the Drosophila larval neuromuscular junction. Robust neural activity did not result in any or noteworthy fluorescence changes when Flash-Pericam, Camgaroo-1, and Camgaroo-2 were expressed. However, calculated on the raw data, fractional fluorescence changes up to 18% were reported by synapto-pHluorin, Yellow Cameleon 2.0, 2.3, and 3.3, Inverse-Pericam, GCaMP1.3, GCaMP1.6, and the troponin C-based calcium sensor TN-L15. The response characteristics of all of these indicators differed considerably from each other, with GCaMP1.6 reporting high rates of neural activity with the largest and fastest fluorescence changes. However, GCaMP1.6 suffered from photobleaching, whereas the fluorescence signals of the double-chromophore indicators were in general smaller but more photostable and reproducible, with TN-L15 showing the fastest rise of the signals at lower activity rates. We show for GCaMP1.3 and YC3.3 that an expanded range of neural activity evoked fairly linear fluorescence changes and a corresponding linear increase in the signal-to-noise ratio (SNR). The expression level of the indicator biased the signal kinetics and SNR, whereas the signal amplitude was independent. The presented data will be useful for in vivo experiments with respect to the selection of an appropriate indicator, as well as for the correct interpretation of the optical signals. PMID- 15888653 TI - Chromatic gain controls in visual cortical neurons. AB - Although the response of a neuron in the visual cortex generally grows nonlinearly with contrast, the spatial tuning of the cell remains stable. This is thought to reflect the activity of a contrast gain control ("normalization") that has very broad tuning on the relevant stimulus dimension. Contrast invariant tuning on a particular dimension is probably necessary for reliable representation of stimuli on that dimension. In the lateral geniculate nucleus (LGN), V1, and V2 of anesthetized macaque, we measured chromatic tuning of neurons at several contrasts to characterize the gain controls and identify cells that might be important for representing color. We estimated separately the chromatic signature of the linear receptive field and that of the gain control. In the LGN, we found normalization in magnocellular cells and cells receiving excitatory S-cone input but not in parvocellular cells or those receiving inhibitory S-cone input. We found normalization in all types of cortical neurons. Among cells that preferred achromatic modulation, or modulation along intermediate directions in color space (making them responsive to both achromatic and chromatic stimuli), normalization was driven by mechanisms tuned to a restricted range of directions in color space, close to achromatic. As a result, chromatic tuning varied with contrast. Among the relatively few cells that strongly preferred chromatic modulation, normalization was driven by mechanisms sensitive to modulation along all directions in color space, especially isoluminant. As a result, chromatic tuning changed little with contrast. To the extent that contrast invariant tuning is important in representing chromaticity, relatively few cortical neurons are involved. PMID- 15888654 TI - Inhibition of conditioned stimulus pathway phosphoprotein 24 expression blocks the reduction in A-type transient K+ current produced by one-trial in vitro conditioning of Hermissenda. AB - Long-term intrinsic enhanced excitability is a characteristic of cellular plasticity and learning-dependent modifications in the activity of neural networks. The regulation of voltage-dependent K+ channels by phosphorylation/dephosphorylation and their localization is proposed to be important in the control of cellular plasticity. One-trial conditioning in Hermissenda results in enhanced excitability in sensory neurons, type B photoreceptors, of the conditioned stimulus pathway. Conditioning also regulates the phosphorylation of conditioned stimulus pathway phosphoprotein 24 (Csp24), a cytoskeletal-related protein containing multiple beta-thymosin-like domains. Recently, it was shown that the downregulation of Csp24 expression mediated by an antisense oligonucleotide blocked the development of enhanced excitability in identified type B photoreceptors after one-trial conditioning without affecting short-term excitability. Here, we show using whole-cell patch recordings that one trial in vitro conditioning applied to isolated photoreceptors produces a significant reduction in the amplitude of the A-type transient K+ current (I(A)) detected 1.5-16 h after conditioning. One-trial conditioning produced a depolarized shift in the steady-state activation curve of I(A) without altering the inactivation curve. The conditioning-dependent reduction in I(A) was blocked by preincubation of the photoreceptors with Csp antisense oligonucleotide. These results provide an important link between Csp24, a cytoskeletal protein, and regulation of voltage-gated ion channels associated with intrinsic enhanced excitability underlying pavlovian conditioning. PMID- 15888655 TI - GABA type-A activity controls its own developmental polarity switch in the maturing retina. AB - In the developing CNS, GABA(A) responses switch from early excitation to late mature inhibition. The developmental factors that induce the polarity switch remain to be unraveled. Here, we bring the first experimental evidence in vivo in the retina that chronic activation of GABA(A) receptors is necessary for the switch to occur and for the chloride extrusion mechanism (through the K+/Cl- cotransporter KCC2) to develop. Using a turtle model and calcium imaging, we investigated how chronic blockade of GABA(A) receptors with bicuculline during the period of the GABAergic polarity switch (from 1 week before hatching until 4 weeks after hatching) influences developmental changes in the patterns of spontaneously generated electrical activity in the retinal ganglion cell (RGC) layer. During that period, spontaneous activity normally switches from propagating waves to stationary patches of coactive cells, until correlated activity completely disappears. These changes in activity patterns coincide with the switch of GABA(A) responses from excitation to inhibition. When GABA(A) receptors are chronically blocked, GABA(A) responses remain excitatory and spontaneous waves keep propagating across the RGC layer. Concomitantly, the developmental upregulation of KCC2 is inhibited on dendritic processes in the inner plexiform layer, suggesting that the intracellular concentration of chloride remains higher, as in younger cells. This study presents the first demonstration in vivo that GABA autoregulates its developmental polarity switch, emphasizing the importance of GABAergic activity in controlling activity patterns in the maturing retina. PMID- 15888656 TI - Distributed neural representation of expected value. AB - Anticipated reward magnitude and probability comprise dual components of expected value (EV), a cornerstone of economic and psychological theory. However, the neural mechanisms that compute EV have not been characterized. Using event related functional magnetic resonance imaging, we examined neural activation as subjects anticipated monetary gains and losses that varied in magnitude and probability. Group analyses indicated that, although the subcortical nucleus accumbens (NAcc) activated proportional to anticipated gain magnitude, the cortical mesial prefrontal cortex (MPFC) additionally activated according to anticipated gain probability. Individual difference analyses indicated that, although NAcc activation correlated with self-reported positive arousal, MPFC activation correlated with probability estimates. These findings suggest that mesolimbic brain regions support the computation of EV in an ascending and distributed manner: whereas subcortical regions represent an affective component, cortical regions also represent a probabilistic component, and, furthermore, may integrate the two. PMID- 15888657 TI - Intracellular calcium regulation by burst discharge determines bidirectional long term synaptic plasticity at the cerebellum input stage. AB - Variations in intracellular calcium concentration ([Ca2+]i) provide a critical signal for synaptic plasticity. In accordance with Hebb's postulate (Hebb, 1949), an increase in postsynaptic [Ca2+]i can induce bidirectional changes in synaptic strength depending on activation of specific biochemical pathways (Bienenstock et al., 1982; Lisman, 1989; Stanton and Sejnowski, 1989). Despite its strategic location for signal processing, spatiotemporal dynamics of [Ca2+]i changes and their relationship with synaptic plasticity at the cerebellar mossy fiber (mf) granule cell (GrC) relay were unknown. In this paper, we report the plasticity/[Ca2+]i relationship for GrCs, which are typically activated by mf bursts (Chadderton et al., 2004). Mf bursts caused a remarkable [Ca2+]i increase in GrC dendritic terminals through the activation of NMDA receptors, metabotropic glutamate receptors (probably acting through IP3-sensitive stores), voltage dependent calcium channels, and Ca2+-induced Ca2+ release. Although [Ca2+]i increased with the duration of mf bursts, long-term depression was found with a small [Ca2+]i increase (bursts <250 ms), and long-term potentiation (LTP) was found with a large [Ca2+]i increase (bursts >250 ms). LTP and [Ca2+]i saturated for bursts >500 ms and with theta-burst stimulation. Thus, bursting enabled a Ca2+-dependent bidirectional Bienenstock-Cooper-Munro-like learning mechanism providing the cellular basis for effective learning of burst patterns at the input stage of the cerebellum. PMID- 15888658 TI - Cyclin-dependent kinase 5 mediates neurotoxin-induced degradation of the transcription factor myocyte enhancer factor 2. AB - Regulation of the process of neuronal death plays a central role both during development of the CNS and in adult brain. The transcription factor myocyte enhancer factor 2 (MEF2) plays a critical role in neuronal survival. Cyclin dependent kinase 5 (Cdk5) mediates neurotoxic effects by phosphorylating and inhibiting MEF2. How Cdk5-dependent phosphorylation reduces MEF2 transactivation activity remained unknown. Here, we demonstrate a novel mechanism by which Cdk5, in conjunction with caspase, inhibits MEF2. Using primary cerebellar granule neuron as a model, our investigation reveals that neurotoxicity induces destabilization of MEF2s in neurons. Destabilization of MEF2 is caused by an increase in caspase-dependent cleavage of MEF2. This cleavage event requires nuclear activation of Cdk5 activity. Phosphorylation by Cdk5 alone is sufficient to promote degradation of MEF2A and MEF2D by caspase-3. In contrast to MEF2A and MEF2D, MEF2C is not phosphorylated by Cdk5 after glutamate exposure and, therefore, resistant to neurotoxin-induced caspase-dependent degradation. Consistently, blocking Cdk5 or enhancing MEF2 reduced toxin-induced apoptosis. These findings define an important regulatory mechanism that for the first time links prodeath activities of Cdk5 and caspase. The convergence of Cdk5 phosphorylation-dependent caspase-mediated degradation of nuclear survival factors exemplified by MEF2 may represent a general process applicable to the regulation of other survival factors under diverse neurotoxic conditions. PMID- 15888659 TI - Functional recovery from desensitization of vanilloid receptor TRPV1 requires resynthesis of phosphatidylinositol 4,5-bisphosphate. AB - Capsaicin and other naturally occurring pungent molecules have long been used as topical analgesics to treat a variety of chronic pain conditions. The analgesic effects of these compounds involve long-term desensitization of nociceptors after strong stimulation. To elucidate the underlying mechanisms, we studied the recovery from desensitization of the vanilloid receptor TRPV1. We showed that prolonged applications of capsaicin led to nearly complete desensitization of the channel and that its functional recovery from desensitization required a high concentration of intracellular ATP. Nonhydrolyzable ATP analogs did not substitute for ATP to promote recovery. Neither inhibition nor activation of protein kinases prevented recovery of the channel from desensitization. In contrast, blockade of lipid kinases, in particular phosphatidylinositol-4-kinase, abolished recovery, as did activation of membrane receptors that stimulate hydrolysis of phosphatidylinositol 4,5-biphosphate (PIP2). Additional experiments using the PIP2-sensitive inward rectifier potassium channel Kir2.1 as a biosensor showed a high degree of temporal correlation between the two channels on both functional suppression after capsaicin stimulation and subsequent recovery. These data suggest that depletion of PIP2 occurs concomitantly with activation of TRPV1 and its replenishment in the membrane determines recovery of the channel from desensitization. In addition to revealing a new role of phosphoinositide signaling in regulation of nociception, our results provide novel insight into the topical mechanisms of the analgesic effects of capsaicin and the strategies to improve its effectiveness. PMID- 15888660 TI - Functional characterization and neuronal modeling of the effects of childhood absence epilepsy variants of CACNA1H, a T-type calcium channel. AB - Sequencing of the T-type Ca2+ channel gene CACNA1H revealed 12 nonsynonymous single nucleotide polymorphisms (SNPs) that were found only in childhood absence epilepsy (CAE) patients. One SNP, G773D, was found in two patients. The present study reports the finding of a third patient with this SNP, as well as analysis of their parents. Because of the role of T-channels in determining the intrinsic firing patterns of neurons involved in absence seizures, it was suggested that these SNPs might alter channel function. The goal of the present study was to test this hypothesis by introducing these polymorphisms into a human Ca(v)3.2a cDNA and then study alterations in channel behavior using whole-cell patch-clamp recording. Eleven SNPs altered some aspect of channel gating. Computer simulations predict that seven of the SNPs would increase firing of neurons, with three of them inducing oscillations at similar frequencies, as observed during absence seizures. Three SNPs were predicted to decrease firing. Some CAE-specific SNPs (e.g., G773D) coexist with SNPs also found in controls (R788C); therefore, the effect of these polymorphisms were studied. The R788C SNP altered activity in a manner that would also lead to enhanced burst firing of neurons. The G773D R788C combination displayed different behavior than either single SNP. Therefore, common polymorphisms can alter the effect of CAE-specific SNPs, highlighting the importance of sequence background. These results suggest that CACNA1H is a susceptibility gene that contributes to the development of polygenic disorders characterized by thalamocortical dysrhythmia, such as CAE. PMID- 15888661 TI - Otx2 regulates subtype specification and neurogenesis in the midbrain. AB - The transcription factor Otx2 is required to determine mesencephalic versus metencephalic (cerebellum/pons) territory during embryogenesis. This function of Otx2 primarily involves positioning and maintaining the mid-hindbrain organizer at the border between midbrain and anterior hindbrain. Otx2 expression is maintained long after this organizer is established. We therefore generated conditional mutants of Otx2 using the Cre/loxP system to study later roles during rostral brain development. For inactivation of Otx2 in neuronal progenitor cells, we crossed Otx2(flox/flox) animals with Nestin-Cre transgenic animals. In Nestin Cre/+; Otx2(flox/flox) embryos, Otx2 activity was lost from the ventral midbrain starting at embryonic day 10.5 (E10.5). In these mutant embryos, the mid hindbrain organizer was properly positioned at E12.5, although Otx2 is absent from the midbrain. Hence, the Nestin-Cre/+; Otx2(flox/flox) animals represent a novel mouse model for studying the role of Otx2 in the midbrain, independently of abnormal development of the mid-hindbrain organizer. Our data demonstrate that Otx2 controls the development of several neuronal populations in the midbrain by regulating progenitor identity and neurogenesis. Dorsal midbrain progenitors ectopically expressed Math1 and generate an ectopic cerebellar-like structure. Similarly, Nkx2.2 ectopic expression ventrally into tegmentum progenitors is responsible for the formation of serotonergic neurons and hypoplasia of the red nucleus in the midbrain. In addition, we discovered a novel role for Otx2 in regulating neurogenesis of dopaminergic neurons. Altogether, these results demonstrate that Otx2 is required from E10.5 onward to regulate neuronal subtype identity and neurogenesis in the midbrain. PMID- 15888662 TI - trkA is expressed in nociceptive neurons and influences electrophysiological properties via Nav1.8 expression in rapidly conducting nociceptors. AB - To test the hypothesis that trkA (the high-affinity NGF receptor) is selectively expressed in nociceptive dorsal root ganglion (DRG) neurons, we examined the intensity of trkA immunoreactivity in single dye-injected rat DRG neurons, the sensory receptor properties of which were identified in vivo with mechanical and thermal stimuli. We provide the first evidence in single identified neurons that strong trkA expression in DRGs is restricted to nociceptive neurons, probably accounting for the profound influence of NGF on these neurons. Furthermore, we demonstrate that trkA expression is as high in rapidly conducting (Aalpha/beta) as in more slowly conducting (Adelta and C) nociceptors. All Aalpha/beta low threshold mechanoreceptors (LTMs) are trkA negative, although weak but detectable trkA is present in some C and Adelta LTMs. NGF can influence electrophysiological properties of DRG neurons, probably by binding to trkA. We found positive correlations for single identified Aalpha/beta (but not C or Adelta) nociceptors between trkA immunocytochemical intensity and electrophysiological properties typical of nociceptors, namely long action potential and afterhyperpolarization durations and large action potential amplitudes. Furthermore, for Aalpha/beta (notCorAdelta) nociceptors, trkA intensity is inversely correlated with conduction velocity. Similar relationships, again only in Aalpha/beta nociceptors, between electrophysiological properties and trkA expression exist for sodium channel Nav1.8 but not Nav1.9 immunoreactivities. These findings suggest that in Aalpha/beta nociceptors, influences of NGF on expression levels of Nav1.8 are related to, and perhaps limited by, expression levels of trkA. This view is supported by a positive correlation between immuno-intensities of trkA and Nav1.8 in A-fiber, but not C-fiber, nociceptors. PMID- 15888663 TI - Truncated prion protein and Doppel are myelinotoxic in the absence of oligodendrocytic PrPC. AB - The cellular prion protein PrP(C) confers susceptibility to transmissible spongiform encephalopathies, yet its normal function is unknown. Although PrP(C) deficient mice develop and live normally, expression of amino proximally truncated PrP(C) (DeltaPrP) or of its structural homolog Doppel (Dpl) causes cerebellar degeneration that is prevented by coexpression of full-length PrP(C). We now report that mice expressing DeltaPrP or Dpl suffer from widespread leukoencephalopathy. Oligodendrocyte-specific expression of full-length PrP(C) under control of the myelin basic protein (MBP) promoter repressed leukoencephalopathy and vastly extended survival but did not prevent cerebellar granule cell (CGC) degeneration. Conversely, neuron-specific PrP(C) expression under control of the neuron-specific enolase (NSE) promoter antagonized CGC degeneration but not leukoencephalopathy. PrP(C) was found in purified myelin and in cultured oligodendrocytes of both wild-type and MBP-PrP transgenic mice but not in NSE-PrP mice. These results identify white-matter damage as an extraneuronal PrP-associated pathology and suggest a previously unrecognized role of PrP(C) in myelin maintenance. PMID- 15888664 TI - Rapid uptake, metabolism, and elimination of inhaled sulfuryl fluoride fumigant by rats. AB - Sulfuryl fluoride (SO(2)F(2)) is a structural fumigant gas used to control drywood termites and wood-boring beetles. The pharmacokinetics and metabolism of inhaled SO(2)F(2) were evaluated in male Fischer-344 rats exposed to 30 or 300 ppm (35)S-labeled SO(2)F(2) for 4 h. Blood, urine and feces were collected during and after the exposures and analyzed for radioactivity, (35)S-labeled fluorosulfate and sulfate, and fluoride (urine and feces only). Selected tissues were collected 7 days post-exposure and analyzed for radioactivity. During and after unlabeled SO(2)F(2) exposures, blood, brain, and kidney were collected and analyzed for fluoride ion. SO(2)F(2) was rapidly absorbed, achieving maximum concentrations of radioactivity in both plasma and red blood cells (RBC) near the end of the 4-h exposure period. Radioactivity was rapidly excreted, mostly via the urine. Seven days post-exposure, small amounts of radioactivity were distributed among several tissues, with the highest concentration detected in respiratory tissues. Radioactivity associated with the RBC remained elevated 7 days post-exposure, and highly perfused tissues had higher levels of radioactivity than other non-respiratory tissues. Radioactivity cleared from plasma and RBC with initial half-lives of 2.5 h after 30 ppm and 1-2.5 h after 300 ppm exposures. The terminal half-life of radioactivity was 2.5-fold longer in RBC than plasma. Based on the radiochemical profiles, there was no evidence of parent (35)SO(2)F(2) in blood. Identification of fluorosulfate and sulfate in blood and urine suggests that SO(2)F(2) is hydrolyzed to fluorosulfate, with release of fluoride, followed by further hydrolysis to sulfate and release of the remaining fluoride. PMID- 15888665 TI - Blood acylpeptide hydrolase activity is a sensitive marker for exposure to some organophosphate toxicants. AB - Acylpeptide hydrolase (APH) unblocks N-acetyl peptides. It is a major serine hydrolase in rat blood, brain, and liver detected by derivatization with (3)H diisopropyl fluorophosphate (DFP) or a biotinylated fluorophosphonate. Although APH does not appear to be a primary target of acute poisoning by organophosphorus (OP) compounds, the inhibitor specificity of this secondary target is largely unknown. This study fills the gap and emphasizes blood APH as a potential marker of OP exposure. The most potent in vitro inhibitors for human erythrocyte and mouse brain APH are DFP (IC(50) 11-17 nM), chlorpyrifos oxon (IC(50) 21-71 nM), dichlorvos (IC(50) 230-560 nM), naled (IC(50) 370-870 nM), and their analogs with modified alkyl substituents. (3)H-diisopropyl fluorophosphate is a potent inhibitor of mouse blood and brain APH in vivo (ED(50) 0.09-0.2 mg/kg and 0.02 0.03 mg/l for ip and vapor exposure, respectively). Mouse blood and brain APH and blood butyrylcholinesterase (BChE) are of similar sensitivity to DFP in vitro and in vivo (ip and vapor exposure), but APH inhibition is much more persistent in vivo (still >80% inhibition after 4 days). The inhibitory potency of OP pesticides in vivo in mice varies from APH selective (dichlorvos, naled, and trichlorfon), to APH and BChE selective (profenofos and tribufos), to ChE selective or nonselective (many commercial insecticides). Sarin administered ip at a lethal dose to guinea pigs inhibits blood acetylcholinesterase and BChE completely but erythrocyte APH only partially. Blood APH activity is therefore a sensitive marker for exposure to some but not all OP pesticides and chemical warfare agents. PMID- 15888666 TI - Comparative microarray analysis of basal gene expression in mouse Hepa-1c1c7 wild type and mutant cell lines. AB - Hepa-1c1c7 wild-type and benzo[a]pyrene-resistant derived mutant cell lines have been used to elucidate pathways and mechanisms involving the aryl hydrocarbon receptor (AhR). However, there has been little focus on other biological processes which may differ between the isolated lines. In this study, mouse cDNA microarrays representing 4858 genes were used to examine differences in basal gene expression between mouse Hepa-1c1c7 wild-type and c1 (truncated Cyp1a1 protein), c4 (AhR nuclear translocator, ARNT, deficient), and c12 (low AhR levels) mutant cell lines. Surprisingly, c1 mutants exhibited the greatest number of gene expression changes compared to wild-type cells, followed by c4 and c12 lines, respectively. Differences in basal gene expression were consistent with cell line specific variations in morphology, mitochondrial activity, and proliferation rate. MTT and direct cell count assays indicate both c4 and c12 mutants exhibit increased proliferative activity when compared to wild-type cells, while the c1 mutants exhibited decreased activity. This study further characterizes Hepa-1c1c7 wild-type and mutant cells and identifies significant differences in biological processes that should be considered when conducting comparative mechanistic studies with these lines. PMID- 15888667 TI - Effects of organochlorine insecticides on MAP kinase pathways in human HaCaT keratinocytes: key role of reactive oxygen species. AB - Organochlorine pesticides (OCs) are reported as potential carcinogens in humans. The aim of this study was to investigate the effects of four OCs (dieldrin, endosulfan, heptachlor, and lindane) on mitogen-activated protein kinase (MAPK) cascades and more specifically to identify the mechanism underlying OC-induced ERK1/2 activation. Organochlorine pesticides increased phosphorylated Raf, MEK1/2, ERK1/2, and c-Jun in human HaCaT cells, but they had no effect on p38 MAPK activation. Moreover, blockade of Raf, MEK1/2, or PKC activation with geldanamycin, U0126, or calphostin C inhibited ERK1/2 phosphorylation, demonstrating a PKC-Raf-MEK1/2 pathway. We also showed that these insecticides induced the production of reactive oxygen species (ROS). Pre-treatment with the antioxidant molecule N-acetyl cysteine sharply decreased the level of phospho ERK1/2 and had no effect on Raf and MEK1/2 activation, suggesting a Raf independent mechanism. This study indicates that OCs strongly activate the ERK1/2 pathway, and it identifies a critical role of ROS in OC-induced ERK activation, probably by stabilizing its phosphorylation. PMID- 15888668 TI - Induction of hepatic peroxisome proliferation by 8-2 telomer alcohol feeding in mice: formation of perfluorooctanoic acid in the liver. AB - The effects of dietary administration of 1H, 1H, 2H, 2H-perfluorodecanol (8-2 telomer alcohol), on peroxisome proliferation in the liver of mice were studied. Male ddY mice were fed on a diet containing 8-2 telomer alcohol at concentrations of 0, 0.025, 0.05, 0.1, and 0.2% (w/w) for 7, 14, 21, and 28 days. These treatments with 8-2 telomer alcohol caused liver enlargement in a dose- and duration-dependent manner. Peroxisome proliferation in the liver of mice was confirmed by electron microscopic examination. Peroxisomal acyl-CoA oxidase was induced by these treatments with 8-2 telomer alcohol in a dose- and time dependent manner. The concentration of perfluorooctanoic acid (PFOA) and related compounds were determined in the liver and plasma, since PFOA had been shown to be a possible metabolite of 8-2 telomer alcohol and to cause significant peroxisome proliferation in rodents. Five metabolites, namely, perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), 2H, 2H-perfluorodecanoic acid (8-2 telomer acid), and two unidentified metabolites, were present in the liver and serum. PFOA was confirmed to be accumulated in the liver of mice following the administration of 8-2 telomer alcohol in a dose- and duration-dependent manner. A linear relationship was observed between the concentration of PFOA and the activity of peroxisomal acyl-CoA oxidase in the liver of mice. These results strongly suggest that PFOA, but not 8-2 telomer alcohol itself, caused peroxisome proliferation in the liver. The present study provided evidence that 8-2 telomer alcohol is converted into PFOA in vivo and that the PFOA formed produces biological effects in the liver of mice. PMID- 15888669 TI - Role of HIF signaling on tumorigenesis in response to chronic low-dose arsenic administration. AB - Trivalent inorganic arsenic (arsenite, arsenic trioxide, As(III)) is a primary contaminant of groundwater supplies worldwide. As(III), marketed as trisenox, is also an FDA-approved agent to treat cancer It has been previously shown by our laboratory that As(III) administered at doses lower than a therapeutic anticancer dose results in an increase in tumor formation and blood vessel density of tumors. In this work it was found that chronic administration of As(III) approaching the EPA action level of 10 ppb, given in the drinking water of mice 5 weeks prior to B16-F10 melanoma implantation, increased the growth rate of primary tumors and the number of metastases to the lung. Further, levels of arsenic in the tumor and lung were found to be much greater than those in the blood and similar to pro-angiogenic As(III) doses. Levels of hypoxia inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) surrounding the blood vessels in the tumors of the As(III)-treated mice were also found to be increased. Exposure of isolated B16-F10 tumor cells to chronic (3 or 7 day) but not acute (4 h) low-dose As(III) was found to increase HIF-1alpha expression and secretion of VEGF. Finally, coadministration of an inhibitor of HIF (YC-1) or a VEGFR-2 kinase inhibitor (SU5416) was found to antagonize the pro angiogenic effects of low-dose As(III). Together, these results suggest that chronic exposure to low-dose As(III) could stimulate growth of tumors through a HIF-dependent stimulation of angiogenesis. PMID- 15888670 TI - In utero and lactational 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure: effects on the prostate and its response to castration in senescent C57BL/6J mice. AB - In utero and lactational 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure inhibits ventral, dorsolateral, and anterior prostate development in C57BL/6 mice. To determine if prostatic abnormalities persist into senescence, mice born to dams given TCDD (5 mug/kg, po) or vehicle on gestation day 13 were examined at 100 and 510 days of age. Half the mice were castrated ten days prior to necropsy in order to assess androgen dependence, while the remaining mice were sham castrated. Effects of TCDD on the dorsolateral and anterior prostate of senescent sham-castrated mice were relatively subtle, whereas the ventral prostate was rudimentary or absent. Castration of vehicle-exposed mice caused far greater reductions in prostate lobe weights, epithelial cell height, and androgen dependent gene expression (MP25 and probasin) in young mice than in senescent ones, while cell proliferation was decreased by castration in young mice and increased in senescence. Responses to castration were similar at 100 days of age in vehicle- and TCDD-exposed mice. At 510 days, however, TCDD-exposed mice were substantially more responsive to castration by most indices than vehicle-exposed mice. These results demonstrate that prostatic androgen dependence in mice declines substantially with age in several key ways, and that in utero and lactational TCDD exposure protects against this decline. Surprisingly, TCDD increased the incidence of cribriform structures in dorsolateral prostate ducts, from 2-3% in vehicle-exposed senescent mice to 16% in sham-castrated and to 7% in castrated senescent mice. Collectively, these results demonstrate that effects of in utero and lactational TCDD exposure on the prostate persist into senescence, and suggest that in utero and lactational TCDD exposure retards the aging process in the prostate. However, because cribriform structures are often considered to be associated with prostate carcinogenesis, these results also suggest that TCDD exposure early in development may increase susceptibility to prostate cancer. PMID- 15888671 TI - Immunotoxic effects of short-term atrazine exposure in young male C57BL/6 mice. AB - The herbicide atrazine (ATR) is a very widely used pesticide; yet the immunotoxicological potential of ATR has not been studied extensively. Our objective was to examine the effect of ATR on selected immune parameters in juvenile mice. ATR (up to 250 mg/kg) was administered by oral gavage for 14 days to one-month-old male C57BL/6 mice. One day, one week, and seven weeks after the last ATR dose, mice were sacrificed, and blood, spleens, and thymuses were collected and processed for cell counting and flow cytometry. Thymus and spleen weights were decreased by ATR, with the thymus being more sensitive than the spleen; this effect was still present at seven days, but not at seven weeks after the last ATR dose. Similarly, organ cellularity was persistently decreased in the thymus and in the spleen, with the splenic, but not thymic cellularity still being depressed at seven weeks post ATR. Peripheral blood leukocyte counts were not affected by ATR. There were also alterations in the cell phenotypes in that ATR exposure decreased all phenotypes in the thymus, with the number of CD4(+)/CD8(+) being affected the least. At the higher doses, the decreases in the thymic T-cell populations were still present one week after the last ATR dose. In the spleen, the CD8(+) were increased and MHC-II(+) and CD19(+) cells were decreased one day after the last ATR dose. Also, ATR treatment decreased the number of splenic naive T helper and T cytotoxic cells, whereas it increased the percentage of highly activated cytotoxic/memory T cells. Interestingly, the proportion of mature splenic dendritic cells (DC; CD11c(high)), was also decreased and it persisted for at least one week, suggesting that ATR inhibited DC maturation. In the circulation, ATR exposure decreased CD4(+) lymphocytes at one day, whereas at seven days after the last ATR dose, in addition to the decrease in CD4(+) lymphocytes, the MHC-II(+) cells were also decreased at the 250 mg/kg dose. Thus, ATR exposure appears to be detrimental to the immune system of juvenile mice by decreasing cellularity and affecting lymphocyte distribution, with certain effects persisting long after exposure has been terminated. PMID- 15888672 TI - Biological databases for plant research. PMID- 15888673 TI - BioMOBY successfully integrates distributed heterogeneous bioinformatics Web Services. The PlaNet exemplar case. AB - The burden of non-interoperability between on-line genomic resources is increasingly the rate-limiting step in large-scale genomic analysis. BioMOBY is a biological Web Service interoperability initiative that began as a retreat of representatives from the model organism database community in September, 2001. Its long-term goal is to provide a simple, extensible platform through which the myriad of on-line biological databases and analytical tools can offer their information and analytical services in a fully automated and interoperable way. Of the two branches of the larger BioMOBY project, the Web Services branch (MOBY S) has now been deployed over several dozen data sources worldwide, revealing some significant observations about the nature of the integrative biology problem; in particular, that Web Service interoperability in the domain of bioinformatics is, unexpectedly, largely a syntactic rather than a semantic problem. That is to say, interoperability between bioinformatics Web Services can be largely achieved simply by specifying the data structures being passed between the services (syntax) even without rich specification of what those data structures mean (semantics). Thus, one barrier of the integrative problem has been overcome with a surprisingly simple solution. Here, we present a non technical overview of the critical components that give rise to the interoperable behaviors seen in MOBY-S and discuss an exemplar case, the PlaNet consortium, where MOBY-S has been deployed to integrate the on-line plant genome databases and analytical services provided by a European consortium of databases and data service providers. PMID- 15888674 TI - The institute for genomic research Osa1 rice genome annotation database. AB - We have developed a rice (Oryza sativa) genome annotation database (Osa1) that provides structural and functional annotation for this emerging model species. Using the sequence of O. sativa subsp. japonica cv Nipponbare from the International Rice Genome Sequencing Project, pseudomolecules, or virtual contigs, of the 12 rice chromosomes were constructed. Our most recent release, version 3, represents our third build of the pseudomolecules and is composed of 98% finished sequence. Genes were identified using a series of computational methods developed for Arabidopsis (Arabidopsis thaliana) that were modified for use with the rice genome. In release 3 of our annotation, we identified 57,915 genes, of which 14,196 are related to transposable elements. Of these 43,719 non transposable element-related genes, 18,545 (42.4%) were annotated with a putative function, 5,777 (13.2%) were annotated as encoding an expressed protein with no known function, and the remaining 19,397 (44.4%) were annotated as encoding a hypothetical protein. Multiple splice forms (5,873) were detected for 2,538 genes, resulting in a total of 61,250 gene models in the rice genome. We incorporated experimental evidence into 18,252 gene models to improve the quality of the structural annotation. A series of functional data types has been annotated for the rice genome that includes alignment with genetic markers, assignment of gene ontologies, identification of flanking sequence tags, alignment with homologs from related species, and syntenic mapping with other cereal species. All structural and functional annotation data are available through interactive search and display windows as well as through download of flat files. To integrate the data with other genome projects, the annotation data are available through a Distributed Annotation System and a Genome Browser. All data can be obtained through the project Web pages at http://rice.tigr.org. PMID- 15888675 TI - MetaCyc and AraCyc. Metabolic pathway databases for plant research. AB - MetaCyc (http://metacyc.org) contains experimentally determined biochemical pathways to be used as a reference database for metabolism. In conjunction with the Pathway Tools software, MetaCyc can be used to computationally predict the metabolic pathway complement of an annotated genome. To increase the breadth of pathways and enzymes, more than 60 plant-specific pathways have been added or updated in MetaCyc recently. In contrast to MetaCyc, which contains metabolic data for a wide range of organisms, AraCyc is a species-specific database containing only enzymes and pathways found in the model plant Arabidopsis (Arabidopsis thaliana). AraCyc (http://arabidopsis.org/tools/aracyc/) was the first computationally predicted plant metabolism database derived from MetaCyc. Since its initial computational build, AraCyc has been under continued curation to enhance data quality and to increase breadth of pathway coverage. Twenty-eight pathways have been manually curated from the literature recently. Pathway predictions in AraCyc have also been recently updated with the latest functional annotations of Arabidopsis genes that use controlled vocabulary and literature evidence. AraCyc currently features 1,418 unique genes mapped onto 204 pathways with 1,156 literature citations. The Omics Viewer, a user data visualization and analysis tool, allows a list of genes, enzymes, or metabolites with experimental values to be painted on a diagram of the full pathway map of AraCyc. Other recent enhancements to both MetaCyc and AraCyc include implementation of an evidence ontology, which has been used to provide information on data quality, expansion of the secondary metabolism node of the pathway ontology to accommodate curation of secondary metabolic pathways, and enhancement of the cellular component ontology for storing and displaying enzyme and pathway locations within subcellular compartments. PMID- 15888676 TI - Databases and information integration for the Medicago truncatula genome and transcriptome. AB - An international consortium is sequencing the euchromatic genespace of Medicago truncatula. Extensive bioinformatic and database resources support the marker anchored bacterial artificial chromosome (BAC) sequencing strategy. Existing physical and genetic maps and deep BAC-end sequencing help to guide the sequencing effort, while EST databases provide essential resources for genome annotation as well as transcriptome characterization and microarray design. Finished BAC sequences are joined into overlapping sequence assemblies and undergo an automated annotation process that integrates ab initio predictions with EST, protein, and other recognizable features. Because of the sequencing project's international and collaborative nature, data production, storage, and visualization tools are broadly distributed. This paper describes databases and Web resources for the project, which provide support for physical and genetic maps, genome sequence assembly, gene prediction, and integration of EST data. A central project Web site at medicago.org/genome provides access to genome viewers and other resources project-wide, including an Ensembl implementation at medicago.org, physical map and marker resources at mtgenome.ucdavis.edu, and genome viewers at the University of Oklahoma (www.genome.ou.edu), the Institute for Genomic Research (www.tigr.org), and Munich Information for Protein Sequences Center (mips.gsf.de). PMID- 15888678 TI - The maize genetics and genomics database. The community resource for access to diverse maize data. AB - The Maize Genetics and Genomics Database (MaizeGDB) serves the maize (Zea mays) research community by making a wealth of genetics and genomics data available through an intuitive Web-based interface. The goals of the MaizeGDB project are 3 fold: to provide a central repository for public maize information; to present the data through the MaizeGDB Web site in a way that recapitulates biological relationships; and to provide an array of computational tools that address biological questions in an easy-to-use manner at the site. In addition to these primary tasks, MaizeGDB team members also serve the community of maize geneticists by lending technical support for community activities, including the annual Maize Genetics Conference and various workshops, teaching researchers to use both the MaizeGDB Web site and Community Curation Tools, and engaging in collaboration with individual research groups to make their unique data types available through MaizeGDB. PMID- 15888677 TI - Genome cluster database. A sequence family analysis platform for Arabidopsis and rice. AB - The genome-wide protein sequences from Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa) spp. japonica were clustered into families using sequence similarity and domain-based clustering. The two fundamentally different methods resulted in separate cluster sets with complementary properties to compensate the limitations for accurate family analysis. Functional names for the identified families were assigned with an efficient computational approach that uses the description of the most common molecular function gene ontology node within each cluster. Subsequently, multiple alignments and phylogenetic trees were calculated for the assembled families. All clustering results and their underlying sequences were organized in the Web-accessible Genome Cluster Database (http://bioinfo.ucr.edu/projects/GCD) with rich interactive and user-friendly sequence family mining tools to facilitate the analysis of any given family of interest for the plant science community. An automated clustering pipeline ensures current information for future updates in the annotations of the two genomes and clustering improvements. The analysis allowed the first systematic identification of family and singlet proteins present in both organisms as well as those restricted to one of them. In addition, the established Web resources for mining these data provide a road map for future studies of the composition and structure of protein families between the two species. PMID- 15888679 TI - Plant protein annotation in the UniProt Knowledgebase. AB - The Swiss-Prot, TrEMBL, Protein Information Resource (PIR), and DNA Data Bank of Japan (DDBJ) protein database activities have united to form the Universal Protein Resource (UniProt) Consortium. UniProt presents three database layers: the UniProt Archive, the UniProt Knowledgebase (UniProtKB), and the UniProt Reference Clusters. The UniProtKB consists of two sections: UniProtKB/Swiss-Prot (fully manually curated entries) and UniProtKB/TrEMBL (automated annotation, classification and extensive cross-references). New releases are published fortnightly. A specific Plant Proteome Annotation Program (http://www.expasy.org/sprot/ppap/) was initiated to cope with the increasing amount of data produced by the complete sequencing of plant genomes. Through UniProt, our aim is to provide the scientific community with a single, centralized, authoritative resource for protein sequences and functional information that will allow the plant community to fully explore and utilize the wealth of information available for both plant and non-plant model organisms. PMID- 15888680 TI - Toward supportive data collection tools for plant metabolomics. AB - Over recent years, a number of initiatives have proposed standard reporting guidelines for functional genomics experiments. Associated with these are data models that may be used as the basis of the design of software tools that store and transmit experiment data in standard formats. Central to the success of such data handling tools is their usability. Successful data handling tools are expected to yield benefits in time saving and in quality assurance. Here, we describe the collection of datasets that conform to the recently proposed data model for plant metabolomics known as ArMet (architecture for metabolomics) and illustrate a number of approaches to robust data collection that have been developed in collaboration between software engineers and biologists. These examples also serve to validate ArMet from the data collection perspective by demonstrating that a range of software tools, supporting data recording and data upload to central databases, can be built using the data model as the basis of their design. PMID- 15888681 TI - Organ-specific expression of Arabidopsis genome during development. AB - The development of complex eukaryotic organisms can be viewed as the selective expression of distinct fractions of the genome in different organs or tissue types in response to developmental and environmental cues. Here, we generated a genome expression atlas of 18 organ or tissue types representing the life cycle of Arabidopsis (Arabidopsis thaliana). We showed that each organ or tissue type had a defining genome expression pattern and that the degree to which organs share expression profiles is highly correlated with the biological relationship of organ types. Further, distinct fractions of the genome exhibited expression changes in response to environmental light among the three seedling organs, despite the fact that they share the same photo-perception and transduction systems. A significant fraction of the genes in the Arabidopsis genome is organized into chromatin domains exhibiting coregulated expression patterns in response to developmental or environmental signals. The knowledge of organ specific expression patterns and their response to the changing environment provides a foundation for dissecting the molecular processes underlying development. PMID- 15888682 TI - Evolutionary divergence of monocot and dicot methyl-CpG-binding domain proteins. AB - The covalent modification of eukaryotic DNA by methylation of the 5' carbon of cytosine residues is frequently associated with transcriptional silencing. In mammals, a potential mechanism for transducing DNA methylation patterns into altered transcription levels occurs via binding of methyl-CpG-binding domain (MBD) proteins. Mammalian MBD-containing proteins bind specifically to methylated DNA and recruit chromatin-modifying complexes containing histone deacetylase activities. Sequence similarity searches reveal the presence of multiple proteins in plants containing a putative MBD. Outside of the MBD itself, there is no sequence relationship between plant and mammalian MBD proteins. The plant MBD proteins can be divided into eight classes based on sequence similarity and phylogenetic analyses of sequences obtained from two complete genomes (rice [Oryza sativa] and Arabidopsis [Arabidopsis thaliana]) and from maize (Zea mays). Two classes of MBD proteins are only represented in dicot species. The striking divergence of plant and animal MBD-containing proteins is in stark contrast to the amino acid conservation of DNA methyltransferases across plants, animals, and fungi. This observation suggests the possibility that while plants and mammals have retained similar mechanisms for the establishment and maintenance of DNA methylation patterns, they may have evolved distinct mechanisms for the interpretation of these patterns. PMID- 15888683 TI - Large-scale identification of expressed sequence tags involved in rice and rice blast fungus interaction. AB - To better understand the molecular basis of the defense response against the rice blast fungus (Magnaporthe grisea), a large-scale expressed sequence tag (EST) sequencing approach was used to identify genes involved in the early infection stages in rice (Oryza sativa). Six cDNA libraries were constructed using infected leaf tissues harvested from 6 conditions: resistant, partially resistant, and susceptible reactions at both 6 and 24 h after inoculation. Two additional libraries were constructed using uninoculated leaves and leaves from the lesion mimic mutant spl11. A total of 68,920 ESTs were generated from 8 libraries. Clustering and assembly analyses resulted in 13,570 unique sequences from 10,934 contigs and 2,636 singletons. Gene function classification showed that 42% of the ESTs were predicted to have putative gene function. Comparison of the pathogen challenged libraries with the uninoculated control library revealed an increase in the percentage of genes in the functional categories of defense and signal transduction mechanisms and cell cycle control, cell division, and chromosome partitioning. In addition, hierarchical clustering analysis grouped the eight libraries based on their disease reactions. A total of 7,748 new and unique ESTs were identified from our collection compared with the KOME full-length cDNA collection. Interestingly, we found that rice ESTs are more closely related to sorghum (Sorghum bicolor) ESTs than to barley (Hordeum vulgare), wheat (Triticum aestivum), and maize (Zea mays) ESTs. The large cataloged collection of rice ESTs in this study provides a solid foundation for further characterization of the rice defense response and is a useful public genomic resource for rice functional genomics studies. PMID- 15888685 TI - Abiotic stress and plant genome evolution. Search for new models. PMID- 15888684 TI - FPC Web tools for rice, maize, and distribution. AB - Many clone-based physical maps have been built with the FingerPrinted Contig (FPC) software, which is written in C and runs locally for fast and flexible analysis. If the maps were viewable only from FPC, they would not be as useful to the whole community since FPC must be installed on the user machine and the database downloaded. Hence, we have created a set of Web tools so users can easily view the FPC data and perform salient queries with standard browsers. This set includes the following four programs: WebFPC, a view of the contigs; WebChrom, the location of the contigs and genetic markers along the chromosome; WebBSS, locating user-supplied sequence on the map; and WebFCmp, comparing fingerprints. For additional FPC support, we have developed an FPC module for BioPerl and an FPC browser using the Generic Model Organism Project (GMOD) genome browser (GBrowse), where the FPC BioPerl module generates the data files for input into GBrowse. This provides an alternative to the WebChrom/WebFPC view. These tools are available to download along with documentation. The tools have been implemented for both the rice (Oryza sativa) and maize (Zea mays) FPC maps, which both contain the locations of clones, markers, genetic markers, and sequenced clone (along with links to sites that contain additional information). PMID- 15888687 TI - Trafficking of normal stem cells and metastasis of cancer stem cells involve similar mechanisms: pivotal role of the SDF-1-CXCR4 axis. AB - The alpha-chemokine stromal-derived factor (SDF)-1 and the G-protein-coupled seven-span transmembrane receptor CXCR4 axis regulates the trafficking of various cell types. In this review, we present the concept that the SDF-1-CXCR4 axis is a master regulator of trafficking of both normal and cancer stem cells. Supporting this is growing evidence that SDF-1 plays a pivotal role in the regulation of trafficking of normal hematopoietic stem cells (HSCs) and their homing/retention in bone marrow. Moreover, functional CXCR4 is also expressed on nonhematopoietic tissue-committed stem/progenitor cells (TCSCs); hence, the SDF-1-CXCR4 axis emerges as a pivotal regulator of trafficking of various types of stem cells in the body. Furthermore, because most if not all malignancies originate in the stem/progenitor cell compartment, cancer stem cells also express CXCR4 on their surface and, as a result, the SDF-1-CXCR4 axis is also involved in directing their trafficking/metastasis to organs that highly express SDF-1 (e.g., lymph nodes, lungs, liver, and bones). Hence, we postulate that the metastasis of cancer stem cells and trafficking of normal stem cells involve similar mechanisms, and we discuss here the common molecular mechanisms involved in these processes. Finally, the responsiveness of CXCR4+ normal and malignant stem cells to an SDF-1 gradient may be regulated positively/primed by several small molecules related to inflammation which enhance incorporation of CXCR4 into membrane lipid rafts, or may be inhibited/blocked by small CXCR4 antagonist peptides. Consequently, strategies aimed at modulating the SDF-1-CXCR4 axis could have important clinical applications both in regenerative medicine to deliver normal stem cells to the tissues/organs and in clinical hematology/oncology to inhibit metastasis of cancer stem cells. PMID- 15888686 TI - Iron-sulfur cluster biogenesis in chloroplasts. Involvement of the scaffold protein CpIscA. AB - The chloroplast contains many iron (Fe)-sulfur (S) proteins for the processes of photosynthesis and nitrogen and S assimilation. Although isolated chloroplasts are known to be able to synthesize their own Fe-S clusters, the machinery involved is largely unknown. Recently, a cysteine desulfurase was reported in Arabidopsis (Arabidopsis thaliana; AtCpNifS) that likely provides the S for Fe-S clusters. Here, we describe an additional putative component of the plastid Fe-S cluster assembly machinery in Arabidopsis: CpIscA, which has homology to bacterial IscA and SufA proteins that have a scaffold function during Fe-S cluster formation. CpIscA mRNA was shown to be expressed in all tissues tested, with higher expression level in green, photosynthetic tissues. The plastid localization of CpIscA was confirmed by green fluorescent protein fusions, in vitro import, and immunoblotting experiments. CpIscA was cloned and purified after expression in Escherichia coli. Addition of CpIscA significantly enhanced CpNifS-mediated in vitro reconstitution of the 2Fe-2S cluster in apo-ferredoxin. During incubation with CpNifS in a reconstitution mix, CpIscA was shown to acquire a transient Fe-S cluster. The Fe-S cluster could subsequently be transferred by CpIscA to apo-ferredoxin. We propose that the CpIscA protein serves as a scaffold in chloroplast Fe-S cluster assembly. PMID- 15888690 TI - Images in clinical medicine. Wernicke's encephalopathy. PMID- 15888688 TI - Human-serum matrix supports undifferentiated growth of human embryonic stem cells. AB - One of the most frequently used matrices for feeder-free growth of undifferentiated human embryonic stem cells (hESCs) is Matrigel, which supports attachment and growth of undifferentiated hESCs in the presence of mouse embryonic fibroblast-conditioned medium. Unfortunately, application of Matrigel or medium conditioned by mouse embryonic feeder cells is not ideal for potential medical application of hESCs because xenogeneic pathogens can be transmitted through culture conditions. We demonstrate here that human serum as matrix and medium conditioned by differentiated hESCs reduce exposure of hESCs to animal ingredients and provide a safer direction toward completely animal-free conditions for application, handling, and understanding of hESC biology. At the same time, hESCs grown under these conditions maintain all hESC features after prolonged culture, including the developmental potential to differentiate into representative tissues of all three embryonic germ layers, unlimited and undifferentiated proliferative ability, and maintenance of normal karyotype. PMID- 15888689 TI - Sequestration and synthesis: the source of insulin in cell clusters differentiated from murine embryonic stem cells. AB - The source of insulin released from insulin-releasing cell clusters (IRCCs) differentiated from embryonic stem cells remains unclear. Rajagopal et al. have suggested that IRCCs do not synthesize but secrete insulin that had been absorbed from media during the multistep protocol. We report here further data relevant to this controversy. No radioisotopic labeling of insulin was observed when IRCCs were incubated in a medium containing 35S-cysteine. Less than 1% of the extra cellular stoichiometric C-peptide equivalent to insulin was secreted during glucose stimulation. However, intracellular immunostaining and immunogold labeling were both positive for C-peptide. Finally, a mass balance calculation showed that simple equilibration of IRCCs by Fickian diffusion from media accounted for at most 4% of secreted insulin. These findings and further analysis of the results of others suggest that the mechanism of insulin secretion by IRCCs is a combination of sequestration and de novo synthesis. PMID- 15888691 TI - Notes to the class--first day. PMID- 15888692 TI - MSL--medicine as a second language. PMID- 15888693 TI - On needless words. PMID- 15888694 TI - Autoimmune folate deficiency and the rise and fall of "horror autotoxicus". PMID- 15888695 TI - Heart-rate profile during exercise as a predictor of sudden death. AB - BACKGROUND: Changes in heart rate during exercise and recovery from exercise are mediated by the balance between sympathetic and vagal activity. Since alterations in the neural control of cardiac function contribute to the risk of sudden death, we tested the hypothesis that among apparently healthy persons, sudden death is more likely to occur in the presence of abnormal heart-rate profiles during exercise and recovery. METHODS: A total of 5713 asymptomatic working men (between the ages of 42 and 53 years), none of whom had clinically detectable cardiovascular disease, underwent standardized graded exercise testing between 1967 and 1972. We examined data on the subjects' resting heart rates, the increase in rate from the resting level to the peak exercise level, and the decrease in rate from the peak exercise level to the level one minute after the termination of exercise. RESULTS: During a 23-year follow-up period, 81 subjects died suddenly. The risk of sudden death from myocardial infarction was increased in subjects with a resting heart rate that was more than 75 beats per minute (relative risk, 3.92; 95 percent confidence interval, 1.91 to 8.00); in subjects with an increase in heart rate during exercise that was less than 89 beats per minute (relative risk, 6.18; 95 percent confidence interval, 2.37 to 16.11); and in subjects with a decrease in heart rate of less than 25 beats per minute after the termination of exercise (relative risk, 2.20; 95 percent confidence interval, 1.02 to 4.74). After adjustment for potential confounding variables, these three factors remained strongly associated with an increased risk of sudden death, with a moderate but significantly increased risk of death from any cause but not of nonsudden death from myocardial infarction. CONCLUSIONS: The heart-rate profile during exercise and recovery is a predictor of sudden death. PMID- 15888696 TI - Circulating osteoblast-lineage cells in humans. AB - BACKGROUND: Although current evidence suggests that only a minuscule number of osteoblast-lineage cells are present in peripheral blood, we hypothesized that such cells circulate but that their concentration has been vastly underestimated owing to the use of assays that required adherence to plastic. We further reasoned that the concentration of these cells is elevated during times of increased bone formation, such as during pubertal growth. METHODS: We used flow cytometry with antibodies to bone-specific proteins to identify circulating osteoblast-lineage cells in 11 adolescent males and 11 adult males (mean [+/-SD] age, 14.5+/-0.7 vs. 37.7+/-7.6 years). Gene expression and in vitro and in vivo bone-forming assays were used to establish the osteoblastic lineage of sorted cells. RESULTS: Cells positive for osteocalcin and cells positive for bone specific alkaline phosphatase were detected in the peripheral blood of adult subjects (1 to 2 percent of mononuclear cells). There were more than five times as many cells positive for osteocalcin in the circulation of adolescent boys (whose markers of bone formation were clearly increased as a result of pubertal growth) as compared with adult subjects (P<0.001). The percentage of cells positive for osteocalcin correlated with markers of bone formation. Sorted osteocalcin-positive cells expressed osteoblastic genes, formed mineralized nodules in vitro, and formed bone in an in vivo transplantation assay. Increased values were also found in three adults with recent fractures. CONCLUSIONS: Osteoblast-lineage cells circulate in physiologically significant numbers, correlate with markers of bone formation, and are markedly higher during pubertal growth; therefore, they may represent a previously unrecognized circulatory component to the process of bone formation. PMID- 15888697 TI - Decreased histone deacetylase activity in chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterized by chronic airway inflammation that is greater in patients with advanced disease. We asked whether there is a link between the severity of disease and the reduction in histone deacetylase (HDAC) activity in the peripheral lung tissue of patients with COPD of varying severity. HDAC is a key molecule in the repression of production of proinflammatory cytokines in alveolar macrophages. METHODS: HDAC activity and histone acetyltransferase (HAT) activity were determined in nuclear extracts of specimens of surgically resected lung tissue from nonsmokers without COPD, patients with COPD of varying severity, and patients with pneumonia or cystic fibrosis. Alveolar macrophages from nonsmokers, smokers, and patients with COPD and bronchial-biopsy specimens from nonsmokers, healthy smokers, patients with COPD, and those with mild asthma were also examined. Total RNA extracted from lung tissue and macrophages was used for quantitative reverse-transcriptase polymerase-chain-reaction assay of HDAC1 through HDAC8 and interleukin-8. Expression of HDAC2 protein was quantified with the use of Western blotting. Histone-4 acetylation at the interleukin-8 promoter was evaluated with the use of a chromatin immunoprecipitation assay. RESULTS: Specimens of lung tissue obtained from patients with increasing clinical stages of COPD had graded reductions in HDAC activity and increases in interleukin-8 messenger RNA (mRNA) and histone-4 acetylation at the interleukin-8 promoter. The mRNA expression of HDAC2, HDAC5, and HDAC8 and expression of the HDAC2 protein were also lower in patients with increasing severity of disease. HDAC activity was decreased in patients with COPD, as compared with normal subjects, in both the macrophages and biopsy specimens, with no changes in HAT activity, whereas HAT activity was increased in biopsy specimens obtained from patients with asthma. Neither HAT activity nor HDAC activity was changed in lung tissue from patients with cystic fibrosis or pneumonia. CONCLUSIONS: Patients with COPD have a progressive reduction in total HDAC activity that reflects the severity of the disease. PMID- 15888698 TI - Radical prostatectomy versus watchful waiting in early prostate cancer. AB - BACKGROUND: In 2002, we reported the initial results of a trial comparing radical prostatectomy with watchful waiting in the management of early prostate cancer. After three more years of follow-up, we report estimated 10-year results. METHODS: From October 1989 through February 1999, 695 men with early prostate cancer (mean age, 64.7 years) were randomly assigned to radical prostatectomy (347 men) or watchful waiting (348 men). The follow-up was complete through 2003, with blinded evaluation of the causes of death. The primary end point was death due to prostate cancer; the secondary end points were death from any cause, metastasis, and local progression. RESULTS: During a median of 8.2 years of follow-up, 83 men in the surgery group and 106 men in the watchful-waiting group died (P=0.04). In 30 of the 347 men assigned to surgery (8.6 percent) and 50 of the 348 men assigned to watchful waiting (14.4 percent), death was due to prostate cancer. The difference in the cumulative incidence of death due to prostate cancer increased from 2.0 percentage points after 5 years to 5.3 percentage points after 10 years, for a relative risk of 0.56 (95 percent confidence interval, 0.36 to 0.88; P=0.01 by Gray's test). For distant metastasis, the corresponding increase was from 1.7 to 10.2 percentage points, for a relative risk in the surgery group of 0.60 (95 percent confidence interval, 0.42 to 0.86; P=0.004 by Gray's test), and for local progression, the increase was from 19.1 to 25.1 percentage points, for a relative risk of 0.33 (95 percent confidence interval, 0.25 to 0.44; P<0.001 by Gray's test). CONCLUSIONS: Radical prostatectomy reduces disease-specific mortality, overall mortality, and the risks of metastasis and local progression. The absolute reduction in the risk of death after 10 years is small, but the reductions in the risks of metastasis and local tumor progression are substantial. PMID- 15888699 TI - Autoantibodies to folate receptors in the cerebral folate deficiency syndrome. AB - In infantile-onset cerebral folate deficiency, 5-methyltetrahydrofolate (5MTHF) levels in the cerebrospinal fluid are low, but folate levels in the serum and erythrocytes are normal. We examined serum specimens from 28 children with cerebral folate deficiency, 5 of their mothers, 28 age-matched control subjects, and 41 patients with an unrelated neurologic disorder. Serum from 25 of the 28 patients and 0 of 28 control subjects contained high-affinity blocking autoantibodies against membrane-bound folate receptors that are present on the choroid plexus. Oral folinic acid normalized 5MTHF levels in the cerebrospinal fluid and led to clinical improvement. Cerebral folate deficiency is a disorder in which autoantibodies can prevent the transfer of folate from the plasma to the cerebrospinal fluid. PMID- 15888700 TI - Cystic fibrosis. PMID- 15888701 TI - Images in clinical medicine. Scrotal cancer. PMID- 15888702 TI - Case records of the Massachusetts General Hospital. Case 14-2005. A 38-year-old man with fever and blurred vision. PMID- 15888703 TI - The fate of circulating osteoblasts. PMID- 15888704 TI - COPD unwound. PMID- 15888705 TI - Matrix metalloproteases and tumor invasion. PMID- 15888706 TI - Implantable cardioverter-defibrillators. PMID- 15888707 TI - N-terminal pro-B-type natriuretic peptide and mortality in coronary heart disease. PMID- 15888708 TI - Expedited treatment of sex partners. PMID- 15888709 TI - Angiotensin II type 1-receptor activating antibodies in renal-allograft rejection. PMID- 15888710 TI - Bites of the brown recluse spider. PMID- 15888711 TI - Antibody response to aerosolized transgenic human alpha1-antitrypsin. PMID- 15888712 TI - A biomechanical comparison of posterior cruciate ligament reconstructions using single- and double-bundle tibial inlay techniques. AB - BACKGROUND: The efficacy of using a double-bundle versus single-bundle graft for posterior cruciate ligament reconstruction has not been demonstrated. HYPOTHESIS: A double-bundle graft restores knee kinematics better than a single-bundle graft does in tibial inlay PCL reconstructions. STUDY DESIGN: Controlled laboratory study. METHODS: Eight cadaveric knees were subjected to 6 cycles from a 40-N anterior reference point to a 100-N posterior translational force at 10 degrees , 30 degrees , 60 degrees , and 90 degrees of flexion. Testing was performed for the intact and posterior cruciate deficient knee as well as for both reconstructed conditions. Achilles tendons, divided into 2 equal sections, were prepared as both single-bundle and double-bundle grafts. Both grafts were employed in the same knee, and the order of graft reconstruction was randomized. RESULTS: There were no statistical differences in translation between the intact state and either of the reconstructions (P > .05) or between either of the reconstructions at any flexion angle (P > .05). CONCLUSION: No differences in translation between the 2 graft options were identified. CLINICAL RELEVANCE: The use of a double-bundle graft may not offer any advantages over a single-bundle graft for tibial inlay posterior cruciate reconstructions. PMID- 15888713 TI - Proximal rectus femoris avulsions in national football league kickers: a report of 2 cases. PMID- 15888714 TI - Biomechanical comparison of the bioabsorbable RetroScrew system, BioScrew XtraLok with stress equalization tensioner, and 35-mm Delta Screws for tibialis anterior graft-tibial tunnel fixation in porcine tibiae. AB - BACKGROUND: Achieving effective soft tissue graft-tibial tunnel fixation remains problematic. HYPOTHESIS: No differences would exist for tibialis anterior graft tibial tunnel fixation when comparing the RetroScrew System (20-mm retrograde screw, 17-mm antegrade screw), the 35-mm tapered Delta Screw (manual tensioning), and the 35-mm BioScrew XtraLok (applied using an instrumented tensioner). STUDY DESIGN: Controlled laboratory study. METHODS: Porcine tibiae (apparent bone mineral density, 1.3 g/cm(2)) and human tendon allografts were divided into 3 matched groups of 6 specimens each before cyclic (500 cycles, 50-250 N, 1 Hz) and load-to-failure (20 mm/min) tests. RESULTS: The BioScrew XtraLok (210.9 +/- 54.9 N/mm) and the 35-mm Delta Screw (224.3 +/- 43.7 N/mm) displayed superior stiffness to the RetroScrew System (114.1 +/- 23.3 N/mm) (P = .0004) during cyclic testing. The BioScrew XtraLok (1.0 +/- 0.2 mm) and the Delta Screw (0.9 +/ 0.2 mm) also displayed less displacement during cyclic testing than the RetroScrew System (1.8 +/- 0.5 mm) (P = .001). During load-to-failure testing, the BioScrew XtraLok withstood greater loads (1436.3 +/- 331.3 N) (P = .001) and displayed greater stiffness (323.6 +/- 56.8 N/mm) (P = .002) than the 35-mm Delta Screw (load, 1042.2 +/- 214.4 N; stiffness, 257.2 +/- 22.2 N/mm) and the RetroScrew System (load, 778.7 +/- 177.5 N; stiffness, 204.4 +/- 52.9 N/mm). CONCLUSION: The BioScrew XtraLok with instrumented tensioning displayed superior fixation to the RetroScrew System and the 35-mm Delta Screw applied with manual tensioning. CLINICAL RELEVANCE: The BioScrew XtraLok may provide superior soft tissue graft-tibial tunnel fixation. Further in vitro studies using human tissue and in vivo clinical studies are needed. PMID- 15888715 TI - Early screw fixation versus casting in the treatment of acute Jones fractures. AB - BACKGROUND: There is considerable variability in the literature concerning the optimal treatment of acute Jones fractures. HYPOTHESIS: Early surgical fixation of acute Jones fractures will result in shorter times to union and return to athletics compared with cast treatment. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 1. METHODS: Eighteen patients were randomized to cast treatment, and 19 patients were randomized to screw fixation. Success of treatment and the times to union and return to sports were calculated for each patient. RESULTS: Mean follow-up was 25.3 months (range, 15-42 months). Eight of 18 (44%) in the cast group were considered treatment failures: 5 nonunions, 1 delayed union, and 2 refractures. One of 19 patients in the surgery group was considered a treatment failure. For the surgery group, the median times to union and return to sports were 7.5 and 8.0 weeks, respectively. For the cast group, the median times were 14.5 and 15.0 weeks, respectively. The Mann-Whitney test showed a statistically significant difference between the groups in both parameters, with P < 001. CONCLUSION: There is a high incidence (44%) of failure after cast treatment of acute Jones fractures. Early screw fixation results in quicker times to union and return to sports compared with cast treatment. PMID- 15888716 TI - Effectiveness of a neuromuscular and proprioceptive training program in preventing anterior cruciate ligament injuries in female athletes: 2-year follow up. AB - BACKGROUND: Among female athletes it has not been established whether a neuromuscular and proprioceptive sports-specific training program will consistently reduce the incidence of anterior cruciate ligament injuries. PURPOSE: To determine whether a neuromuscular and proprioceptive performance program was effective in decreasing the incidence of anterior cruciate ligament injury within a select population of competitive female youth soccer players. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: In 2000, 1041 female subjects from 52 teams received a sports-specific training intervention in a prospective non-randomized trial. The control group consisted of the remaining 1905 female soccer players from 95 teams participating in the same league who were age and skill matched. In the 2001 season, 844 female athletes from 45 teams were enrolled in the study, with 1913 female athletes (from 112 teams) serving as the age- and skill-matched controls. All subjects were female soccer players between the ages of 14 and 18 and participated in either their traditional warm up or a sports-specific training intervention before athletic activity over a 2 year period. The intervention consisted of education, stretching, strengthening, plyometrics, and sports-specific agility drills designed to replace the traditional warm-up. RESULTS: During the 2000 season, there was an 88% decrease in anterior cruciate ligament injury in the enrolled subjects compared to the control group. In year 2, during the 2001 season, there was a 74% reduction in anterior cruciate ligament tears in the intervention group compared to the age- and skill-matched controls. CONCLUSION: Using a neuromuscular training program may have a direct benefit in decreasing the number of anterior cruciate ligament injuries in female soccer players. PMID- 15888717 TI - Long-term failure of thermal shrinkage for laxity of the anterior cruciate ligament. AB - BACKGROUND: Appropriate treatment for anterior cruciate ligament laxity, owing to partial tears of the native ligament or lax reconstruction, is unclear. Studies suggest that a significant percentage of these untreated tears may progress to complete tears or the patient may develop additional injuries to the meniscus or articular cartilage. Shrinkage of the ligament or graft using thermal energy has been proposed as a solution for this problem. PURPOSE: To evaluate the long-term results using thermal energy to shrink laxity of the anterior cruciate ligament. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Nineteen patients with partial tears of the anterior cruciate ligament (n = 14) or stretched anterior cruciate ligament grafts (n = 5) underwent thermal shrinkage treatment. Fourteen were available for follow-up at 1 and 5 years. At 1 year, there were 12 partial tears and 2 grafts available for evaluation. At 5 years, there were 10 partial tears and 4 grafts. Preoperative, postoperative, and intraoperative stability testing was performed using the KT-1000 arthrometer. Clinical results were evaluated using the Cincinnati and Lysholm scoring systems. RESULTS: Intraoperative shrinkage averaged 2.12 mm (17%, P < .0001). At 1 year, 12 of the 14 patients remained stable (86%) with a negative Lachman test result and mean KT 1000 arthrometer maximum side-to-side score of 1.29 mm (P < .001). The 2 failed shrinkages were partial tears of the native anterior cruciate ligament. For these 12 patients, Cincinnati scores improved from 53 to 89 (P < .0001), and Lysholm scores improved from 55 to 89 (P < .0003). At 5-year follow-up, 11 of 13 patients had gone on to complete failure (85%, P < .002; 8/9 partial tears and 3/4 grafts). CONCLUSION: Thermal shrinkage provides short-term benefit in the treatment of anterior cruciate ligament laxity but leads to catastrophic failure in the majority of patients at long-term follow-up. We can no longer recommend this procedure for the treatment of anterior cruciate ligament laxity. PMID- 15888718 TI - Revision surgery for exertional anterior compartment syndrome of the lower leg: technique, findings, and results. AB - BACKGROUND: Recurrent symptoms or failure after fasciotomy for exertional anterior compartment syndrome is not uncommon. HYPOTHESIS: Symptoms from high compartment pressures can be secondary to involvement of the entire compartment or to localized constrictions from postsurgical fibrosis, as well as to entrapment of the superficial peroneal nerve. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Eighteen patients who underwent revision surgery for exertional anterior compartment syndrome were available for follow-up. All were athletes who had either a failure or a recurrence of symptoms at a mean of 23.5 months (range, 8-54 months) after the index fasciotomy. Pressure measurements using a slit catheter at rest, at 1 minute postexercise, and at 5 minutes postexercise were performed in 2 places within the compartment: in the area of the previous incision and in the proximal muscle belly of the tibialis anterior. Surgical technique consisted of a 2-incision approach with partial fasciectomy, exploration and decompression of the superficial peroneal nerve, and excision of all fibrotic tissue. An objective examination and a comprehensive subjective questionnaire previously described were performed at a mean follow-up of 42 months (range, 22-67 months). RESULTS: Sixty percent of patients had abnormal pressures only in a localized area, whereas 40% had high pressures throughout the compartment. Eight of 18 (44%) patients had symptoms, signs, and surgical findings of entrapment of the superficial peroneal nerve. At follow-up, 72% of patients had a satisfactory outcome (5 excellent, 8 good), and 28% had an unsatisfactory outcome for intense running sports (4 fair, 1 poor), although 3 patients with the fair results reported improvement with low-level activity. All 8 patients with documented peroneal nerve entrapment had a satisfactory outcome. CONCLUSION: Symptoms from high pressures can be secondary to involvement of the entire compartment or localized to a certain area from postsurgical fibrosis. Pressure measurements should be performed in at least 2 separate areas. PMID- 15888719 TI - Chronic anterior midtibial stress fractures in athletes treated with reamed intramedullary nailing. AB - BACKGROUND: A chronic anterior midtibial stress fracture is a serious, difficult to-treat injury that can adversely affect an athlete's career. HYPOTHESIS: The use of a reamed intramedullary nail for a chronic anterior tibial stress fracture is a safe and effective treatment for an athlete. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Seven collegiate-level athletes with 11 chronic anterior midtibial stress fractures were treated with reamed intramedullary nailing between 1997 and 2000. These patients were followed for a mean duration of 17 months. RESULTS: The mean age of the patients at the time of stress fracture diagnosis was 17 years. Seven of the fractures occurred in male athletes, whereas 4 occurred in female athletes. All patients had failed nonoperative treatment, including rest, activity modification, use of an orthosis, and low-intensity ultrasound stimulation, for a minimum of 4 months. Patients had experienced symptoms for a mean duration of 12 months. Clinical and radiological union occurred at a mean of 2.7 and 3 months, respectively. The mean duration for return to sports after surgery was 4 months. At last follow-up, all patients had full range of motion at the knee and ankle joints and were satisfied with the results. One patient developed bursitis at the tibial nail insertion site that was resolved with a steroid injection. Another patient sustained a traumatic fracture of the distal tibia 1 year after intramedullary nailing of the tibial stress fracture. This fracture healed with nonoperative treatment. No other complications were observed. CONCLUSION: Intramedullary nailing of the tibia for chronic stress fracture has a high union rate, allows for a low complication rate, and allows for an early return to competitive sports. This procedure is an excellent alternative treatment for those fractures that have failed nonsurgical treatment. PMID- 15888720 TI - Neuromuscular and biomechanical adaptations of patients with isolated deficiency of the posterior cruciate ligament. AB - BACKGROUND: Functional adaptations of patients with posterior cruciate ligament deficiency (grade II) are largely unknown despite increased recognition of this injury. HYPOTHESIS: Posterior cruciate ligament-deficient subjects (grade II, 6- to 10-mm bilateral difference in posterior translation) will present with neuromuscular and biomechanical adaptations to overcome significant mechanical instability during gait and drop-landing tasks. STUDY DESIGN: Controlled laboratory study. METHODS: Bilateral comparisons were made among 10 posterior cruciate ligament-deficient subjects using radiographic, instrumented laxity, and range of motion examinations. Biomechanical and neuromuscular characteristics of the involved limb of the posterior cruciate ligament-deficient subjects were compared to their uninvolved limb and to 10 matched control subjects performing gait and drop-landing tasks. RESULTS: Radiographic (15.3 +/- 2.9 to 5.6 +/- 3.7 mm; P = .008) and instrumented laxity (6.3 +/- 2.0 to 1.4 +/- 0.5 mm; P < .001) examinations demonstrated significantly greater posterior displacement of the involved knee within the posterior cruciate ligament-deficient group. The posterior cruciate ligament-deficient group had a significantly decreased maximum knee valgus moment and greater vertical ground reaction force at midstance during gait compared to the control group. During vertical landings, the posterior cruciate ligament-deficient group demonstrated a significantly decreased vertical ground reaction force loading rate. All other analyses reported no significant differences within or between groups. CONCLUSION: Posterior cruciate ligament deficient subjects demonstrate minimal biomechanical and neuromuscular differences despite significant clinical laxity. CLINICAL RELEVANCE: The findings of this study indicate that individuals with grade II posterior cruciate ligament injuries are able to perform gait and drop-landing activities similar to a control group without surgical intervention. PMID- 15888721 TI - Effects of radiofrequency energy on human articular cartilage: an analysis of 5 systems. AB - BACKGROUND: Previous radiofrequency work has not rigidly controlled energy application to the articular cartilage, giving uncertain results published to date. HYPOTHESIS: At minimal settings, radiofrequency probes cause cell death in measurable areas when applied to human articular cartilage. STUDY DESIGN: Controlled laboratory study. METHODS: Simulating operating room conditions, 5 commercially available radiofrequency probes were attached to a customized jig to standardize a minimal contact pressure of each probe tip to 2.0 g. Keeping all variables the same, probes were placed on specific points of fresh grade II human cartilage with treatment times of 1 and 3 seconds at the manufacturer's recommended settings. Grade III cartilage was also tested with a treatment time of 3 seconds, and grade II cartilage was studied with the probe held 1 mm off the cartilage surface. Cartilage was blindly analyzed by confocal microscopy using a live/dead cell viability assay to determine the extent of cell death. RESULTS: Radiofrequency probes produced significant cellular death in the form of a half circle into the cartilage to variable depths. For treatment times of 1 and 3 seconds, cell death measurements ranged from 404 to 539 mum and 1034 to 1283 mum, respectively. One probe failed to show any effect, with minimal evidence of cell death or cartilage smoothing. When probes were kept a 1.0-mm distance above the cartilage, no cell death or cartilage smoothing was noted. Radiofrequency treatment of grade III cartilage penetrated to the subchondral bone. There was no statistically significant difference between the damage caused by monopolar and bipolar probes when tested under these rigidly controlled conditions. CONCLUSION: These results showed significant cellular death at these minimal conditions to the underlying chondrocytes with radiofrequency probes. Surgeons using this technology need to be aware of the power and dangerous potential these probes can have on articular cartilage. PMID- 15888723 TI - Failure mode of suture anchors as a function of insertion depth. AB - BACKGROUND: Surgeons can control not only the angle but also the depth of suture anchor placement during arthroscopic rotator cuff repair, although the tendency may be to place suture anchors on the deep side to avoid damage from prominent anchor eyelets. However, little information is available regarding possible effects of suture anchor depth on construct failure mechanisms. HYPOTHESIS: Anchor depth affects the mode of suture failure with physiologically relevant cyclic loads. STUDY DESIGN: Controlled laboratory study. METHODS: Metallic screw in suture anchors loaded with No. 2 braided polyester sutures were inserted into the bovine infra-spinatus footprint with the eyelet proud, standard, or deep. Sutures were hand tied to create a closed loop. Constructs were cyclically loaded from 10 to 90 N and, if still intact at 500 cycles, taken to ultimate failure (maximum load). RESULTS: When clinical failure was defined as greater than 3-mm construct elongation, anchors placed with the eyelet deep experienced statistically earlier clinical failure via cutting of the suture through the bone (P < .02). However, anchors placed at this level did not experience catastrophic failure during cyclic loading. The standard and proud anchors experienced 3 mm of elongation at a greater number of cycles, but the suture material degraded at the anchor eyelet, and a majority of these constructs broke during cyclic physiologic loading. At failure testing, the deep anchors had a significantly increased failure load (164 N) compared to standard (133 N) (P < .04) and proud (113 N) anchors (P < .005). CONCLUSION: Varying the depth of suture anchor insertion changes the mechanical properties and mode of failure of suture anchor constructs. CLINICAL RELEVANCE: Surgeons should be aware of the effects of suture anchor depth and abrasive eyelet wear on construct failure during arthroscopic rotator cuff repair. PMID- 15888722 TI - Characteristics of water skiing-related and wakeboarding-related injuries treated in emergency departments in the United States, 2001-2003. AB - BACKGROUND: Water skiing and wakeboarding are popular sports with high potential for injury due to rapid boat acceleration, lack of protective gear, and waterway obstacles. However, trends in water skiing- and wakeboarding-related injuries in the United States have not been described using national data. HYPOTHESIS: The number of injuries, injury diagnoses, and body regions injured vary by sport. STUDY DESIGN: Descriptive epidemiology study. METHODS: Data regarding water skiing- and wakeboarding-related injuries presenting to 98 hospital emergency departments in the United States between January 1, 2001, and December 31, 2003, were extracted from the National Electronic Injury Surveillance System. Data included demographics, injury diagnosis, and body region injured. RESULTS: Data were collected for 517 individuals with water skiing-related injuries and 95 individuals with wakeboarding-related injuries. These injuries represent an estimated 23 460 water skiing- and 4810 wakeboarding-related injuries treated in US emergency departments in 2001 to 2003. Head injuries represented the largest percentage of injuries for wakeboarders (28.8% of all injuries) and the smallest percentage for water skiers (4.3%) (P < .01; relative risk [95% confidence interval], 6.73 [3.89-11.66]). Analysis of injury diagnosis was consistent as wakeboarders had significantly more traumatic brain injuries (12.5% of all injuries) than did water skiers (2.4%) (P < .05; relative risk [95% confidence interval], 5.27 [2.21-12.60]). Strains or sprains were the leading injury diagnoses for water skiing (36.3% of all injuries), and the majority (55.7%) were to the lower extremity. Lacerations were the most common diagnoses for wakeboarders (31.1% of all injuries), and the majority (59.6%) were to the face. CONCLUSION: The analyses of water skiing- and wakeboarding-related injuries treated in US emergency departments in 2001 to 2003 highlight the differences in injury patterns for these 2 sports. The substantial number of head and facial injuries among wakeboarders underscores the need for research on the potential role of helmets or other protective gear to reduce these common injuries. PMID- 15888724 TI - The effects of low-intensity ultrasound on medial collateral ligament healing in the rabbit model. AB - BACKGROUND: Ruptured medial collateral ligaments are capable of healing over time, but biomechanical and biochemical properties remain inferior to normal tissue. Low-intensity ultrasound may improve healing. HYPOTHESIS: Medial collateral ligaments treated with ultrasound will demonstrate superior healing. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty-one late-adolescent male rabbits underwent bilateral ligament transection. One ligament from each rabbit received ultrasound treatment every other day for 6 total treatments. Contralateral ligaments received sham treatments. After 3 or 6 weeks, ligaments were evaluated biomechanically and assayed for collagen concentration and the relative proportions of types I and III collagen. RESULTS: Areas of sonicated specimens were significantly larger (10.6% +/- 4.90%) at 6 weeks. Ultimate load (39.5% +/- 17.0%), ultimate displacement (24.5% +/- 8.0%), and energy absorption (69.1% +/- 22.0%) were significantly higher for sonicated specimens at 6 weeks. No significant biomechanical differences were observed at 3 weeks. The relative proportion of type I collagen was significantly higher in sonicated ligaments at 3 weeks (8.61% +/- 4.0%) and 6 weeks (6.91% +/- 3.0%). No significant differences in collagen concentration were observed at either 3 or 6 weeks. CONCLUSION: Subtle improvement with ultrasound treatment may be apparent by 3 weeks after injury, suggested by increased proportion of type I collagen. Ultrasound appears to improve some structural properties and to modestly increase scar cross sectional area and type I collagen present at 6 weeks after injury in this model. CLINICAL RELEVANCE: Ultrasound treatments after ligament injury may facilitate earlier return to activities and decrease risk of reinjury. PMID- 15888725 TI - Fifty years of research on the intelligence of deaf and hard-of-hearing children: a review of literature and discussion of implications. AB - In 1965, McCay Vernon drove a stake through the heart of the long-established "truth" that deaf people were inferior to hearing people. Launched by Aristotle, emboldened by the 1880 Conference of Milan, and reiterated in the twentieth century through the biased research of many psychologists, this falsehood persisted until the publication of this classic review paper. Vernon succinctly spotlights biases in IQ assessment of deaf children resulting from improper testing methods, research participant sampling, even the experience level of the evaluators themselves. Brief and scholarly, the paper had enormous impact not only on future research regarding cognition and deaf people but on clinical practice as well. Within this paper, insights are evident which Vernon has continued to elucidate throughout his long career. He was arguably the first psychologist to view the deaf population as a heterogeneous one, noting how various hearing loss etiologies differentially affect cognition and other psychological characteristics. His later research, notably that which focused on rubella, deafblind, and deaf forensic populations, has been similarly pioneering. McCay Vernon's extensive professional impact stems not only from his prolific, readily applied research work but also from his generous and vigorous activities as a teacher, a mentor, and advisor. - Robert Q. Pollard, Jr. PMID- 15888726 TI - ZAC, LIT1 (KCNQ1OT1) and p57KIP2 (CDKN1C) are in an imprinted gene network that may play a role in Beckwith-Wiedemann syndrome. AB - Loss of genomic imprinting is involved in a number of developmental abnormalities and cancers. ZAC is an imprinted gene expressed from the paternal allele of chromosome 6q24 within a region known to harbor a tumor suppressor gene for several types of neoplasia. p57(KIP2) (CDKN1C) is a maternally expressed gene located on chromosome 11p15.5 which encodes a cyclin-dependent kinase inhibitor that may also act as a tumor suppressor gene. Mutations in ZAC and p57KIP2 have been implicated in transient neonatal diabetes mellitus (TNDB) and Beckwith Wiedemann syndrome, respectively. Patients with these diseases share many characteristics. Here we show that mouse Zac1 and p57Kip2 have a strikingly similar expression pattern. ZAC, a sequence-specific DNA-binding protein, binds within the CpG island of LIT1 (KCNQ1OT1), a paternally expressed, anti-sense RNA thought to negatively regulate p57(KIP2) in cis. ZAC induces LIT1 transcription in a methylation-dependent manner. Our data suggest that ZAC may regulate p57(KIP2) through LIT1, forming part of a novel signaling pathway regulating cell growth. Mutations in ZAC may, therefore, contribute to Beckwith-Wiedemann syndrome. Furthermore, we find changes in DNA methylation at the LIT1 putative imprinting control region in two patients with TNDB. PMID- 15888727 TI - Nucleoside diphosphate kinase from Mycobacterium tuberculosis cleaves single strand DNA within the human c-myc promoter in an enzyme-catalyzed reaction. AB - The reason for secretion of nucleoside diphosphate kinase (NdK), an enzyme involved in maintaining the cellular pool of nucleoside triphosphates in both prokaryotes and eukaryotes, by Mycobacterium tuberculosis is intriguing. We recently observed that NdK from M.tuberculosis (mNdK) localizes within nuclei of HeLa and COS-1 cells and also nicks chromosomal DNA in situ (A. K. Saini, K. Maithal, P. Chand, S. Chowdhury, R. Vohra, A. Goyal, G. P. Dubey, P. Chopra, R. Chandra, A. K. Tyagi, Y. Singh and V. Tandon (2004) J. Biol. Chem., 279, 50142 50149). In the current study, using a molecular beacon approach, we demonstrate that the mNdK catalyzes the cleavage of single strand DNA. It displays Michaelis Menten kinetics with a kcat/K(M) of 9.65 (+/-0.88) x 10(6) M(-1) s(-1). High affinity (K(d) approximately K(M) of approximately 66 nM) and sequence-specific binding to the sense strand of the nuclease hypersensitive region in the c-myc promoter was observed. This is the first study demonstrating that the cleavage reaction is also enzyme-catalyzed in addition to the enzymatic kinase activity of multifunctional NdK. Using our approach, we demonstrate that GDP competitively inhibits the nuclease activity with a K(I) of approximately 1.9 mM. Recent evidence implicates mNdK as a potent virulence factor in tuberculosis owing to its DNase-like activity. In this context, our results demonstrate a molecular mechanism that could be the basis for assessing in situ DNA damage by secretory mNdK. PMID- 15888728 TI - Regulation of ribonucleotide reductase M2 expression by the upstream AUGs. AB - Ribonucleotide reductase catalyzes a rate-limiting reaction in DNA synthesis by converting ribonucleotides to deoxyribonucleotides. It consists of two subunits and the small one, M2 (or R2), plays an essential role in regulating the enzyme activity and its expression is finely controlled. Changes in the M2 level influence the dNTP pool and, thus, DNA synthesis and cell proliferation. M2 gene has two promoters which produce two major mRNAs with 5'-untranslated regions (5' UTRs) of different lengths. In this study, we found that the M2 mRNAs with the short (63 nt) 5'-UTR can be translated with high efficiency whereas the mRNAs with the long (222 nt) one cannot. Examination of the long 5'-UTR revealed four upstream AUGs, which are in the same reading frame as the unique physiological translation initiation codon. Further analysis demonstrated that these upstream AUGs act as negative cis elements for initiation at the downstream translation initiation codon and their inhibitory effect on M2 translation is eIF4G dependent. Based on the findings of this study, we conclude that the expression of M2 is likely regulated by fine tuning the translation from the mRNA with a long 5'-UTR during viral infection and during the DNA replication phase of cell proliferation. PMID- 15888729 TI - Common patterns in type II restriction enzyme binding sites. AB - Restriction enzymes are among the best studied examples of DNA binding proteins. In order to find general patterns in DNA recognition sites, which may reflect important properties of protein-DNA interaction, we analyse the binding sites of all known type II restriction endonucleases. We find a significantly enhanced GC content and discuss three explanations for this phenomenon. Moreover, we study patterns of nucleotide order in recognition sites. Our analysis reveals a striking accumulation of adjacent purines (R) or pyrimidines (Y). We discuss three possible reasons: RR/YY dinucleotides are characterized by (i) stronger H bond donor and acceptor clusters, (ii) specific geometrical properties and (iii) a low stacking energy. These features make RR/YY steps particularly accessible for specific protein-DNA interactions. Finally, we show that the recognition sites of type II restriction enzymes are underrepresented in host genomes and in phage genomes. PMID- 15888730 TI - Cigarette smoke inhibits hamster oocyte pickup by increasing adhesion between the oocyte cumulus complex and oviductal cilia. AB - Previous research has shown that oocyte cumulus complex (OCC) pickup rate is inhibited in hamsters when oviducts and OCCs are simultaneously exposed to cigarette smoke solutions, independent of any effect on ciliary beat frequency. The purpose of this research was to determine whether smoke solutions caused a change in adhesion between the OCC and infundibulum of the oviduct and to determine whether a change in adhesion could account for decreased OCC pickup rate. OCC pickup rate and adhesion were measured before and after infundibula or OCCs recovered from acute in vitro exposures to mainstream and sidestream whole, gas, or particulate smoke solutions. Ciliary beat frequency was also measured on infundibula. Overall, smoke solutions decreased oocyte pickup rate 40% to 80% below control levels and increased adhesion 52% to 91% above control levels when infundibula were pretreated. A change in adhesion was observed in cases for which decreased OCC pickup rate could not be explained by a change in ciliary beat frequency. OCC pickup rate decreased 20% to 35% below control levels and adhesion increased 39% to 54% above control levels when OCCs were pretreated. These data show not only that cigarette smoke causes an increase in OCC adhesion to the oviduct, but also that there is a correlation between increased adhesion and decreased OCC pickup rate. The results also show that the both the OCC and oviduct are targets of cigarette smoke. The oviduct is more sensitive to the adverse effects of smoke; however, this may be caused by a combined impact on mechanisms involved in both adhesion and ciliary function. PMID- 15888731 TI - Transport of IgG across the blood-luminal barrier of the male reproductive tract of the rat and the effect of estradiol administration on reabsorption of fluid and IgG by the epididymal ducts. AB - In rats immunized systemically with tetanus toxoid the concentration of specific anti-tetanus-toxoid-specific IgG in fluid from the rete testis and cauda epididymidis were respectively 0.6% and 1.4% the concentration in blood serum. The extratesticular duct system reabsorbed 97% of the IgG and 99% of the fluid leaving the rete, but estradiol administration affected the site of reabsorption. In untreated rats, the ductuli efferentes reabsorbed 94% of the IgG and 96% of the fluid leaving the rete, whereas estradiol-treated rats reabsorbed 83% of the IgG and 86% of the fluid, and the ductus epididymidis fully compensated for these different effects of estradiol on the ductuli efferentes. The concentrations of IgG in secretions of the seminal vesicles and prostate gland were lower (0.1% and 0.3% respectively of the titers in blood serum) than in fluids from the extratesticular ducts, and were not affected by the administration of estradiol. RT-PCR showed that Fcgrt (neonatal Fc receptor, also known as FcRn) is expressed in the reproductive ducts, where IgG is probably transported across epithelium, being particularly strong in the ductuli efferentes (where most IgG was reabsorbed) and distal caput epididymidis. It is concluded that IgG enters the rete testis and is concentrated only 2.5-fold along the extratesticular duct system, unlike spermatozoa, which are concentrated 95-fold. Further, the ductus epididymidis can recognize and compensate for changes in function of the ductuli efferentes. PMID- 15888732 TI - Spermadhesin AQN1 is a candidate receptor molecule involved in the formation of the oviductal sperm reservoir in the pig. AB - Sperm are stored in the isthmic region of the oviduct under conditions that maintain viability and suppress early capacitation steps until ovulation occurs. The initial contact between sperm and oviductal epithelium is mediated by carbohydrate-protein interactions. In the pig, the carbohydrate recognition system has been shown to involve oligomannosyl structures. The spermadhesins AWN and AQN1 are the dominant porcine carbohydrate-binding sperm proteins. The objective of this study was to demonstrate that AQN1 contributes to sperm binding to the oviductal epithelium. AQN1 showed a broad carbohydrate-binding pattern as it recognizes both alpha- and beta-linked galactose as well as Manalpha1 3(Manalpha1-6)Man structures, whereas AWN bound only the galactose species. Binding of ejaculated sperm to oviductal epithelium was inhibited by addition of AQN1 but not by AWN. Mannose-binding sites were localized over the rostral region of the sperm head. Flow cytometry showed that, under capacitating conditions, the population of live sperm was shifted within 30 min toward an increase in the proportion of cells with low mannose- and high galactose-binding. The loss of mannose-binding sites was accompanied by the loss of AQN1 in sperm extracts and the significant reduction in the sperm-oviduct binding. The oviductal epithelium was shown by GNA-lectin histochemistry and by SDS-PAGE and lectin blotting of the apical membrane fraction to express mannose components that could be recognized by AQN1. These results demonstrate that the sperm lectin AQN1 fulfils the criteria for an oviduct receptor in the pig and may play a role in the formation of the oviductal sperm reservoir. PMID- 15888733 TI - Ovulation-inducing factor in the seminal plasma of alpacas and llamas. AB - Studies were conducted to document the existence of an ovulation-inducing factor in the seminal plasma of alpacas (experiment 1) and llamas (experiment 2) and to determine if the effect is mediated via the pituitary (experiment 3). In experiment 1, female alpacas (n = 14 per group) were given alpaca seminal plasma or saline intramuscularly or by intrauterine infusion. Only alpacas that were given seminal plasma i.m. ovulated (13/ 14, 93%; P < 0.01). In experiment 2, ovulation was detected in 9/10 (90%) llamas at a mean of 29.3 +/- 0.7 h after seminal plasma treatment. Plasma progesterone concentrations were maximal by Day 9 and were at nadir by Day 12 posttreatment. In experiment 3, female llamas were given llama seminal plasma, GnRH, or saline i.m., and ovulation was detected in 6/6, 5/ 6, and 0/6 llamas, respectively (P < 0.001). Treatment was followed by a surge (P < 0.01) in plasma LH concentration beginning 15 min and 75 min after treatment with GnRH and seminal plasma, respectively. Plasma LH remained elevated longer in the seminal plasma group (P < 0.05) and had not yet declined to pretreatment levels after 8 h. Compared with the GnRH group, corpus luteum tended to grow longer and to a greater diameter (P = 0.1) and plasma progesterone concentration was twice as high in the seminal plasma group (P < 0.01). Results document the existence of a potent factor in the seminal plasma of alpacas and llamas that elicited a surge in circulating concentrations of LH and induced an ovulatory and luteotropic response. PMID- 15888734 TI - A placenta-specific enhancer of the human syncytin gene. AB - The cis- and trans-acting factors that are critical for placenta-specific expression of the human syncytin gene are unknown. We identified a 146-base pair (bp) region of the 5'-flanking region of the human syncytin gene from nt-294 to 148 that is essential for basal gene expression in human BeWo and JEG3 choriocarcinoma cell lines but not in hepatoblastoma and kidney cell lines. Ligation of the 146-bp fragment to a SV40 promoter or a human beta-globin minimal promoter markedly enhanced promoter activity in the placenta cells but not in the liver and kidney cells. DNase I footprint assays indicated that nuclear extracts from BeWo cells but not HepG2 cells protected four regions (FP1-FP4) of the 146 bp fragment. Site-directed mutagenesis of an SP1-binding site in FP3 and a GATA binding site in FP4 significantly repressed promoter activity in the placenta cells. Overexpression of SP1 (Sp1 transcription factor) and GATA2 (GATA binding protein 2) and GATA3 induced syncytin promoter activity but had little or no effect on the activities of syncytin promoter fragments containing mutations in the SP1- and GATA-binding sites. GATA2 and -3 mRNA levels increased markedly during spontaneous in vitro differentiation of human cytotrophoblast cells when the cytotrophoblast cells fused to form a syncytium. These findings strongly suggest that the 146-bp region of the 5'-flanking region (nt-294/-148) of the human syncytin gene acts as a placenta-specific enhancer. Binding of SP1 and GATA family members to this enhancer is critical for cell-specific expression of the syncytin gene. PMID- 15888735 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of locally recurrent or metastatic breast cancer (MBC). PMID- 15888736 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of epithelial ovarian carcinoma. PMID- 15888737 TI - ESMO Minimum Clinical Recommendations for diagnosis, adjuvant treatment and follow-up of colon cancer. PMID- 15888738 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of advanced colorectal cancer. PMID- 15888739 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of rectal cancer. PMID- 15888740 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of gastric cancer. PMID- 15888741 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of pancreatic cancer. PMID- 15888742 TI - ESMO Minimal Clinical Recommendations for diagnosis, treatment and follow-up of esophageal cancer. PMID- 15888743 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of non-small-cell lung cancer (NSCLC). PMID- 15888744 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of small-cell lung cancer (SCLC). PMID- 15888745 TI - Minimum Clinical Recommendations for diagnosis, treatment and follow-up of malignant pleural mesothelioma. PMID- 15888746 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of prostate cancer. PMID- 15888747 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of mixed or non-seminomatous germ cell tumors (NSGCT). PMID- 15888748 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of testicular seminoma. PMID- 15888749 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of invasive bladder cancer. PMID- 15888750 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of multiple myeloma. PMID- 15888751 TI - ESMO Minimum Clinical Recommendations for the diagnosis, treatment and follow-up of acute myeloblastic leukemia (AML) in adult patients. PMID- 15888752 TI - ESMO Minimum Clinical Recommendations: assuring common standards of care. PMID- 15888753 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of chronic lymphocytic leukemia. PMID- 15888754 TI - ESMO Minimum Clinical Recommendations for the diagnosis, treatment and follow-up of chronic myelogenous leukemia (CML). PMID- 15888755 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of Hodgkin's disease. PMID- 15888756 TI - Minimum Clinical Recommendations for diagnosis, treatment and follow-up of newly diagnosed follicular lymphoma. PMID- 15888757 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of newly diagnosed large cell non-Hodgkin's lymphoma. PMID- 15888758 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of relapsed large cell non-Hodgkin's lymphoma. PMID- 15888759 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of squamous cell carcinoma of the head and neck (SCCHN). PMID- 15888760 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of malignant glioma. PMID- 15888761 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of cutaneous malignant melanoma. PMID- 15888762 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of soft tissue sarcomas. PMID- 15888763 TI - ESMO Minimum Clinical Recommendations for diagnosis, adjuvant treatment and follow-up of primary breast cancer. PMID- 15888764 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of osteosarcoma. PMID- 15888765 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of Ewing's sarcoma of bone. PMID- 15888766 TI - ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of cancers of unknown primary site (CUP). PMID- 15888767 TI - ESMO Minimum Clinical Recommendations for prophylaxis of chemotherapy-induced nausea and vomiting (NV). PMID- 15888768 TI - ESMO recommendations for the application of hematopoietic growth factors. PMID- 15888769 TI - ESMO Minimum Clinical Recommendations for the management of cancer pain. PMID- 15888771 TI - The time to eradicate gastric cancer is now. AB - Worldwide gastric cancer remains one of the most common cancers, killing upwards of one million people each year. While the molecular pathogenesis remains unclear, infection with the bacterium Helicobacter pylori is considered a "necessary but not sufficient" cause, not surprisingly as gastric cancer has long been known to be associated with atrophic gastritis. Eradication of H pylori is expected to virtually eliminate gastric cancer and H pylori associated peptic ulcer within approximately 40 years and thus reduce overall mortality. In the USA, the incidence of gastric cancer in the general population is low, reflecting the change in the pattern of gastritis from atrophic to non-atrophic and in the low and decreasing prevalence of H pylori infection in the middle and upper classes. However, the plan for eradication of this important pathogen must be considered within the context of the prevalence and outcome within specific populations. PMID- 15888770 TI - Guidelines for the management of patients with pancreatic cancer periampullary and ampullary carcinomas. PMID- 15888772 TI - Gastro-oesophageal reflux disease: symptoms, erosions, and Barrett's--what is the interplay? PMID- 15888773 TI - Screening for early gastric cancer. PMID- 15888774 TI - Creeping fat in Crohn's disease: travelling in a creeper lane of research? PMID- 15888775 TI - Are we studying the correct state of the stellate cell to elucidate mechanisms of chronic pancreatitis? PMID- 15888776 TI - Prognostic influence of Barrett's oesophagus and Helicobacter pylori infection on healing of erosive gastro-oesophageal reflux disease (GORD) and symptom resolution in non-erosive GORD: report from the ProGORD study. AB - BACKGROUND: Adequacy of acid suppression is a critical factor influencing healing in gastro-oesophageal reflux disease (GORD). The European prospective study ProGORD was set up to determine the endoscopic and symptomatic progression of GORD over five years under routine care, after initial acid suppression with esomeprazole. We report on factors influencing endoscopic healing and symptom resolution during the acute treatment phase. METHODS: Patients with symptoms suggestive of GORD underwent endoscopy and biopsies were obtained from the oesophagus for diagnosis of abnormalities, including Barrett's oesophagus (BO). Data from 6215 patients were included in the "intention to treat" analysis, 3245 diagnosed as having erosive reflux disease (ERD) and 2970 non-erosive reflux disease (NERD). ERD patients were treated with esomeprazole 40 mg for 4-8 weeks for endoscopic healing while NERD patients received 20 mg for 2-4 weeks for resolution of heartburn symptoms. RESULTS: Endoscopic healing occurred overall in 87.7% of ERD patients although healing was significantly lower in those with more severe oesophagitis (76.9%) and in those with BO (72.4%), particularly in Helicobacter pylori negative BO patients (70.1%). Age, sex, and body mass index appeared to have no significant impact on healing. Complete heartburn resolution was reported by 70.4% of ERD patients and by 64.8% of NERD patients at the last visit. Only H pylori infection had a significant influence on complete heartburn resolution in the NERD group (68.1% and 63.7% for H pylori positive and H pylori negative, respectively; p = 0.03). CONCLUSION: The presence of Barrett's mucosa, as well as severe mucosal damage, exerts a negative impact on healing. H pylori infection had a positive influence on healing in ERD patients with coexistent BO but no influence on those without BO. PMID- 15888777 TI - Long term failure of endoscopic gastroplication (EndoCinch). AB - INTRODUCTION: Endoluminal gastroplication (EndoCinch; Bard) has been introduced as an endoscopic treatment option in gastro-oesophageal reflux disease (GORD) patients with promising short term results. However, little is known about the long term efficacy of endoscopic suturing. The aim of this study was to evaluate prospectively the long term outcome after EndoCinch. PATIENTS AND METHODS: A total of 70 patients treated with EndoCinch at a single referral centre were studied prospectively. All patients were interviewed using a standardised questionnaire regarding their symptoms and medication prior to and 18 months after EndoCinch. In addition, follow up included endoscopy, 24 hour pH monitoring, and oesophageal manometry. RESULTS: The procedure was well tolerated without major short or long term complications. Eighteen months after EndoCinch, 56/70 patients (80%) were considered treatment failures as their heartburn symptoms did not improve or proton pump inhibitor medication exceeded 50% of the initial dose. Endoscopy showed all sutures in situ in 12/70 (17%) patients while no remaining sutures could be detected in 18/70 (26%). In 54 and 50 patients examined, respectively, no significant changes in 24 hour pH monitoring (median pH <4/24 hours, 9.1% v 8.5%; p = 0.82) or lower oesophageal sphincter (LOS) pressure (7.7 v 10.3 mm Hg; p = 0.051) were observed while median LOS length slightly increased (3.0 to 3.2 cm; p<0.05). CONCLUSION: Endoscopic gastroplication (EndoCinch) is a safe and minimally invasive endoscopic treatment for GORD with reasonable short term results. In contrast, long term outcome is disappointing, probably due to suture loss in the majority of patients. Therefore, technical improvements to ensure suture durability are mandatory before endoscopic suturing can evolve as a therapeutic option for GORD treatment. PMID- 15888778 TI - Rare presentation of a common illness. Campylobacter jejuni infection. PMID- 15888779 TI - Risk factors for oesophageal squamous dysplasia in adult inhabitants of a high risk region of China. AB - BACKGROUND: Oesophageal squamous cell carcinoma (OSCC) is a common cancer worldwide and has a very high mortality rate. Squamous dysplasia is the precursor lesion for OSCC and it can be seen during routine endoscopy with Lugol's iodine staining. We aimed to examine the risk factors for squamous dysplasia and determine if a risk model could be constructed which would be useful in selecting apparently healthy subjects for endoscopic screening in a high risk population in Linzhou, People's Republic of China. SUBJECTS AND METHODS: In this cross sectional study, 724 adult volunteers aged 40-65 years were enrolled. All subjects completed a questionnaire regarding potential environmental exposures, received physical and dental examinations, and underwent upper endoscopy with Lugol's iodine staining and biopsy. Subjects were categorised as having or not having histologically proven squamous dysplasia/early cancer. Risk factors for dysplasia were examined using univariate and multivariate logistic regression. The utility of the final multivariate model as a screening tool was assessed using a receiver operating characteristics curve. RESULTS: We found that 230 of 720 subjects (32%) with complete data had prevalent squamous dysplasia. In the final multivariate model, more household members (odds ratio (OR) 1.12/member (95% confidence interval (CI) 0.99, 1.25)), a family history of cancer (OR 1.57 (95% CI 1.13-2.18)), higher systolic blood pressure OR 1.11/10 mm Hg (95% CI 1.03 1.19)), heating the home without a chimney (OR 2.22 (95% CI 1.27-3.86)), and having lost more but not all of your teeth (OR 1.91 for 12-31 teeth lost (95% CI 1.17-3.15)) were associated with higher odds of having dysplasia. Higher household income (OR 0.96/100 RMB (95% CI 0.91-1.00)) was associated with a lower odds of having dysplasia. Although we found several statistically significant associations, the final model had little ability to accurately predict dysplasia status, with maximum simultaneous sensitivity and specificity values of 57% and 54%, respectively. CONCLUSIONS: We found that risk factors for dysplasia were similar to those previously identified as risk factors for OSCC in this population. The final model did a poor job of identifying subjects who had squamous dysplasia. Other methods will need to be developed to triage individuals to endoscopy in this high risk population. PMID- 15888780 TI - Predicting the development of gastric cancer from combining Helicobacter pylori antibodies and serum pepsinogen status: a prospective endoscopic cohort study. AB - BACKGROUND AND AIMS: Helicobacter pylori infection and gastric atrophy are both risk factors for gastric cancer. We aimed to elucidate the natural history of gastric cancer development according to H pylori infection and gastric atrophy status. SUBJECTS AND METHODS: A total of 9293 participants in a mass health appraisal programme were candidates for inclusion in the present prospective cohort study: 6983 subjects revisited the follow up programme. Subjects were classified into four groups according to serological status at initial endoscopy. Group A (n = 3324) had "normal" pepsinogen and were negative for H pylori antibody; group B (n = 2134) had "normal" pepsinogen and were positive for H pylori antibody; group C (n = 1082) had "atrophic" pepsinogen and were positive for H pylori antibody; and group D (n = 443) had "atrophic" pepsinogen and were negative for H pylori antibody. Incidence of gastric cancer was determined by annual endoscopic examination. RESULTS: Mean duration of follow up was 4.7 years and the average number of endoscopic examinations was 5.1. The annual incidence of gastric cancer was 0.04% (95% confidence interval (CI) 0.02-0.09), 0.06% (0.03 0.13), 0.35% (0.23-0.57), and 0.60% (0.34-1.05) in groups A, B, C, and D, respectively. Hazard ratios compared with group A were 1.1 (95% CI 0.4-3.4), 6.0 (2.4-14.5), and 8.2 (3.2-21.5) in groups B, C, and D, respectively. Age, sex, and "group" significantly served as independent valuables by multivariate analysis. CONCLUSIONS: The combination of serum pepsinogen and anti-H pylori antibody provides a good predictive marker for the development of gastric cancer. PMID- 15888781 TI - An unusual complication of Crohn's colitis. PMID- 15888782 TI - Gut mucosal granulocyte activation precedes nitric oxide production: studies in coeliac patients challenged with gluten and corn. AB - BACKGROUND AND AIMS: To elucidate the dynamics of nitric oxide (NO) production induced by rectal gluten challenge and the relation between NO production and mucosal granulocyte activation. SUBJECTS AND METHODS: Release of rectal NO was measured in 13 patients with coeliac disease and in 18 controls before and after rectal wheat gluten challenge. Rectal gas was collected with a rectal balloon using a newly developed instrument/technique, the "mucosal patch technique". The instrument allows simultaneous measurements of concentrations of granulocyte mediators in the rectal mucosa. We measured myeloperoxidase (MPO), eosinophil cationic protein (ECP), and histamine. For comparison, we made similar measurements after corn (maize) gluten challenge. RESULTS: In all coeliac patients rectal NO concentration increased after gluten challenge and reached a peak after 15 hours (mean 9464 (SEM 2393) parts per billion (ppb); range 250 24982). The maximum MPO and ECP increase occurred five hours after challenge. A correlation was found between mucosal MPO and NO production at 15 hours. Six of the patients showed an increase in NO production 15 hours after rectal corn gluten challenge but this was much smaller than after gluten challenge. No increases were seen in the control group after either challenge. CONCLUSION: Mucosal activation of neutrophils and eosinophils precedes pronounced enhancement of mucosal NO production after rectal wheat gluten challenge in patients with coeliac disease. Some of our coeliac patients displayed signs of an inflammatory reaction, as measured by NO and granulocyte markers, after rectal corn gluten challenge. PMID- 15888783 TI - An unusual cause of pyrexia. PMID- 15888784 TI - Reparative properties of a commercial fish protein hydrolysate preparation. AB - BACKGROUND: A partially hydrolysed and dried product of pacific whiting fish is currently marketed as a health food supplement to support "intestinal health". However, there has been only limited scientific study regarding its true biological activity. AIMS: We therefore tested its efficacy in a variety of models of epithelial injury and repair. METHODS: Effects on proliferation were determined using [(3)H] thymidine incorporation into epithelial rat intestinal RIE-1 and human colonic HT29 cells. Effects on restitution (cell migration) were analysed using wounded HT29 monolayers and its ability to influence gastric injury analysed using a rat indomethacin restraint model. Partial characterisation of bioactive agents was performed using mass spectroscopy, high pressure liquid chromatography, and gas chromatography. RESULTS: Both cell proliferation and cell migration were increased by about threefold when added at 1 mg/ml (p<0.01). Gastric injury was reduced by 59% when gavaged at 25 mg/ml (p<0.05), results similar to using the potent cytoprotective agent epidermal growth factor at 12.5 mug/ml. The vast majority of biological activity was soluble in ethanol, with glutamine in its single, di-, and tripeptide forms probably accounting for approximately 40% of the total bioactivity seen. Fatty acid constituents may also have contributed to cell migratory activity. CONCLUSIONS: Fish protein hydrolysate possesses biological activity when analysed in a variety of models of injury and repair and could provide a novel inexpensive approach for the prevention and treatment of the injurious effects of non steroidal anti-inflammatory drugs and other ulcerative conditions of the bowel. Further studies appear justified. PMID- 15888785 TI - Patient defined dichotomous end points for remission and clinical improvement in ulcerative colitis. AB - BACKGROUND AND AIMS: Ulcerative colitis disease activity indices offer good statistical power but small changes in these indices may not be clinically important. There are no validated definitions of remission or of significant improvement for these indices. The use of clinically important end points would strengthen the validity of study outcomes. Our aims were to identify objective end points in standard disease activity indices for remission and for improvement in ulcerative colitis. METHODS: Sixty six consecutive patients with ulcerative colitis provided information about remission status and their disease activity. At a return visit 1-14 months later, these patients provided information about the change in their disease activity, and non-invasive indices were measured. RESULTS: Specific objective end points for determining remission with four standard indices and a quality of life instrument were determined (St Mark's <3.5, ulcerative colitis disease activity index <2.5, simple clinical colitis activity index (SCCAI) <2.5, Seo <120, and inflammatory bowel disease quality of life index (IBDQ) >205). These cut offs also identified patients who met a regulatory definition of remission. Specific objective end points for clinical improvement in two non-invasive indices and a quality of life instrument were determined with good sensitivity and specificity (SCCAI decrease >1.5, Seo decrease >30, IBDQ increase >20). CONCLUSIONS: We found specific cut off values for disease activity indices that identify patients who have significantly improved or achieved remission in an objective, sensitive, and specific manner. These cut offs should help in the interpretation of the outcomes of clinical trials in ulcerative colitis. PMID- 15888786 TI - Production of adiponectin, an anti-inflammatory protein, in mesenteric adipose tissue in Crohn's disease. AB - BACKGROUND AND AIMS: A characteristic feature of Crohn's disease (CD) is mesenteric adipose tissue hypertrophy. Mesenteric adipocytes or specific proteins secreted by them may play a role in the pathogenesis of CD. We recently identified adiponectin as an adipocyte specific protein with anti-inflammatory properties. Here we report on expression of adiponectin in mesenteric adipose tissue of CD patients. METHODS AND RESULTS: Mesenteric adipose tissue specimens were obtained from patients with CD (n = 22), ulcerative colitis (UC) (n = 8) and, for controls, colon carcinoma patients (n = 28) who underwent intestinal resection. Adiponectin concentrations were determined by enzyme linked immunosorbent assay, and adiponectin mRNA levels were determined by real time quantitative reverse transcription-polymerase chain reaction. Tissue concentrations and release of adiponectin were significantly increased in hypertrophied mesenteric adipose tissue of CD patients compared with normal mesenteric adipose tissue of CD patients (p = 0.002, p = 0.040, respectively), UC patients (p = 0.002, p = 0.003), and controls (p<0.0001, p<0.0001). Adiponectin mRNA levels were significantly higher in hypertrophied mesenteric adipose tissue of CD patients than in paired normal mesenteric adipose tissue from the same subjects (p = 0.024). Adiponectin concentrations in hypertrophied mesenteric adipose tissue of CD patients with an internal fistula were significantly lower than those of CD patients without an internal fistula (p = 0.003). CONCLUSIONS: Our results suggest that adipocytes in hypertrophied mesenteric adipose tissue produce and secrete significant amounts of adiponectin, which could be involved in the regulation of intestinal inflammation associated with CD. PMID- 15888787 TI - O(6)-methylguanine methyltransferase in colorectal cancers: detection of mutations, loss of expression, and weak association with G:C>A:T transitions. AB - BACKGROUND AND AIMS: O(6)-methylguanine methyltransferase (MGMT) repairs inappropriately methylated guanine in DNA. MGMT mutations have not previously been reported in cancers, but in colorectal tumours MGMT promoter methylation is common and has been associated with increased G:C>A:T transitions, a high frequency of K-ras mutations, and low level microsatellite instability (MSI low). However, some have suggested that MGMT changes are background or secondary events, with little importance for tumorigenesis. METHODS: We have analysed fresh frozen colorectal cancers and colorectal cancer cell lines for MGMT changes: mutations, allelic loss, and protein expression. RESULTS: Six of 113 cancers harboured somatic missense MGMT mutations, at least three of which probably caused reduced MGMT function and were accompanied by silencing or loss of the wild-type allele. Cancers with pathogenic MGMT mutations tended to harbour G:C>A:T somatic mutations at other loci. Overall, MGMT expression was reduced or lost in more than half of the cancers. We found no association between MGMT expression and the somatic mutation spectrum at APC, beta-catenin, K-ras, or p53, but decreased MGMT expression was weakly associated with the presence of a G:C>A:T change at any one of these loci. Reduced MGMT expression was not however associated with an increased frequency of K-ras mutations or with MSI low. CONCLUSION: In summary, we found that mutation of MGMT contributes to decreased protein function. Our findings provide good evidence to show that MGMT changes, including methylation, are selected rather than background events, at least in some cases. Decreased MGMT expression or function probably has a weak or moderate effect on the mutation spectrum in colorectal cancers. PMID- 15888788 TI - Interval faecal occult blood testing in a colonoscopy based screening programme detects additional pathology. AB - BACKGROUND: Colonoscopic based surveillance is recommended for patients at increased risk of colorectal cancer. The appropriate interval between surveillance colonoscopies remains in debate, as is the "miss rate" for colorectal cancer within such screening programmes. AIMS: The main aim of this study was to determine whether a one-off interval faecal occult blood test (FOBT) facilitates the detection of significant neoplasia within a colonoscopic based surveillance programme. Secondary aims were to determine if invitees were interested in participating in interval screening, and to determine whether interval lesions were missed or whether they developed rapidly since the previous colonoscopy PATIENTS: Patients enrolled in a colonoscopic based screening programme due to a personal history of colorectal neoplasia or a significant family history. METHODS: Patients within the screening programme were invited to perform an immunochemical FOBT (Inform). A positive result was followed by colonoscopy; significant neoplasia was defined as colorectal cancer, adenomas either > or =10 mm or with a villous component, high grade dysplasia, or multiplicity (>/=3 adenomas). Participation rates were determined for age, sex, and socioeconomic subgroups. Colonoscopy recall databases were examined to determine the interval between previous colonoscopy and FOBT offer, and correlations between lesion characteristics and interval time were determined. RESULTS: A total of 785 of 1641 patients invited (47.8%) completed an Inform kit. A positive result was recorded for 57 (7.3%). Fifty two of the 57 test positive patients completed colonoscopy; 14 (1.8% of those completing the FOBT) had a significant neoplastic lesion. These consisted of six colorectal cancers and eight significant adenomas. CONCLUSIONS: A one off immunochemical faecal occult blood test within a colonoscopy based surveillance programme had a participation rate of nearly 50% and appeared to detect additional pathology, especially in patients with a past history of colonic neoplasia. PMID- 15888789 TI - Quality in the technical performance of screening flexible sigmoidoscopy: recommendations of an international multi-society task group. AB - BACKGROUND: Flexible sigmoidoscopy (FS) is a complex technical procedure performed in a variety of settings, by examiners with diverse professional backgrounds, training, and experience. Potential variation in technical quality may have a profound impact on the effectiveness of FS on the early detection and prevention of colorectal cancer. AIM: We propose a set of consensus and evidence based recommendations to assist the development of continuous quality improvement programmes around the delivery of FS for colorectal cancer screening. RECOMMENDATIONS: These recommendations address the intervals between FS examinations, documentation of results, training of endoscopists, decision making around referral for colonoscopy, policies for antibiotic prophylaxis and management of anticoagulation, insertion of the FS endoscope, bowel preparation, complications, the use of non-physicians as FS endoscopists, and FS endoscope reprocessing. For each of these areas, continuous quality improvement targets are recommended, and research questions are proposed. PMID- 15888790 TI - An upper abdominal swelling causing nausea and vomiting. PMID- 15888791 TI - Translocation of p21(Cip1/WAF1) from the nucleus to the cytoplasm correlates with pancreatic myofibroblast to fibroblast cell conversion. AB - BACKGROUND AND AIMS: In the pancreas, myofibroblasts (MFBs) were shown to play an important role in the cellular response during inflammation and injury. However, there is only fragmentary information concerning the fate of these cells in pancreas regeneration and fibrosis development. METHODS: Explant cultures of rat pancreatic tissue were used as a model to follow cellular dynamics and phenotype conversion of pancreatic MFBs in vitro. For detailed biochemical analyses a pancreatic fibroblast cell line (long culture fibroblast (LCF)) was generated from MFBs in a long term culture. Cerulein induced acute pancreatitis and dibutyltin dichloride induced pancreas fibrosis were used as experimental models for acute and chronic fibrogenic reactions, respectively. RESULTS: In the explant culture, pancreatic MFBs which derived from fat storing fibroblastic cells underwent apoptosis or converted again to fibroblasts. The phenotype switch to fibroblasts was associated with translocation of p21(Cip1/WAF1) from the nucleus into the cytoplasm. Molecular analyses in LCFs revealed subsequent binding to and inhibition of the activities of Rho kinase 2 and apoptosis signal regulating kinase 1. In the experimentally established pancreas fibrosis, fibroblasts with cytoplasmic expression of p21(Cip1/WAF1) were distributed throughout fibrotic bands whereas in experimental acute pancreatitis MFBs with nuclear expression of p21(Cip1/WAF1) dominated. CONCLUSIONS: The results indicate that pancreatic MFBs are transient and suggest that intracellular localisation of p21(Cip1/WAF1) can contribute to the phenotype conversion of these cells to fibroblasts in culture and experimental injury. PMID- 15888792 TI - Dietary carbohydrates and glycaemic load and the incidence of symptomatic gall stone disease in men. AB - BACKGROUND: Diets with a high glycaemic response exacerbate the metabolic consequences of the insulin resistance syndrome. Their effects on the incidence of gall stone disease are not clear, particularly in men. METHODS: Dietary information was collected as part of the Health Professionals Follow up Study starting in 1986 using a semiquantitative food frequency questionnaire with follow up until 1998. On biennial questionnaires participants reported new symptomatic gall stone disease, diagnosed by radiology, and whether they had undergone cholecystectomy. RESULTS: During 12 years of follow up, we documented 1810 new cases of symptomatic gall stones. After adjusting for age and other known or suspected risk factors in multivariate models, the relative risk (RR) for the highest compared with the lowest quintile of carbohydrate intake was 1.59 (95% confidence interval (CI) 1.25, 2.02; p for trend = 0.002). The RR for the highest compared with the lowest quintile of dietary glycaemic load was 1.50 (95% CI 1.20, 1.88; p for trend = 0.0008), and 1.18 for dietary glycaemic index (95% CI 1.01, 1.39; p for trend = 0.04). Independent positive associations were also seen for intakes of starch, sucrose, and fructose. CONCLUSIONS: Our findings suggest that a high intake of carbohydrate, glycaemic load, and glycaemic index increases the risk of symptomatic gall stone disease in men. These results add to the concern that low fat high carbohydrate diets may not be an optimal dietary recommendation. PMID- 15888793 TI - ATP8B1 mutations in British cases with intrahepatic cholestasis of pregnancy. AB - BACKGROUND: Intrahepatic cholestasis of pregnancy (ICP) affects approximately 0.7% of pregnancies in the UK and is associated with prematurity, fetal distress, and intrauterine death. Homozygous mutations in the ATP8B1 gene cause cholestasis with a normal serum gamma-glutamyl transpeptidase (gamma-GT), and have been reported in two forms of cholestasis: progressive familial intrahepatic cholestasis type 1 (PFIC1) and benign recurrent intrahepatic cholestasis (BRIC). AIMS: To establish whether mutations in ATP8B1 are associated with ICP in British cases PATIENTS: Sixteen well phenotyped women with ICP without raised gamma-GT were selected for sequence analysis. Subsequently, 182 patients and 120 controls were examined for the presence of the variants detected. METHODS: All coding exons were sequenced in 16 cases. Eight ICP cases, including two women carrying a mutation, were investigated using in vivo hepatic (31)P magnetic resonance spectroscopy (MRS) RESULTS: Two heterozygous ATP8B1 transitions (208G>A and 2599C>T) that resulted in amino acid substitutions were identified; 208G>A was identified in three cases. MRS revealed an increased phosphodiester signal (Mann Whitney U test, p = 0.03) and a decreased phosphomonoester/phosphodiester ratio (p = 0.04) in ICP cases compared with controls. CONCLUSIONS: We were able to demonstrate ATP8B1 mutations in ICP. MRS studies suggest that susceptibility to ICP is associated with a relative rise in biliary phospholipid. These data also suggest that MRS may be used for non-invasive assessment of the liver and biliary constituents in cholestasis. PMID- 15888794 TI - Nuclear factor {kappa}B inactivation in the rat liver ameliorates short term total warm ischaemia/reperfusion injury. AB - BACKGROUND: In hepatic ischaemia/reperfusion injury, activated liver macrophages (Kupffer cells) are dominantly regulated by a transcription factor, nuclear factor kappaB (NFkappaB), with respect to expression of inflammatory cytokines, acute phase response proteins, and cell adhesion molecules. AIMS: We assessed whether inactivation of NFkappaB in the liver could attenuate total hepatic warm ischaemia/reperfusion injury. METHODS: We studied rats with hepatic overexpression of inhibitor kappaBalpha super-repressor (IkappaBalpha SR) caused by a transgene introduced using an adenoviral vector. Hepatic ischaemia/reperfusion injury was induced under warm conditions by total occlusion of hepatoduodenal ligament structures for 20 minutes, followed by reperfusion. Controls included uninfected and control virus (AdLacZ) infected rats. RESULTS: IkappaBalpha SR was overexpressed in Kupffer cells as well as in hepatocytes, blocking nuclear translocation of NFkappaB (p65) into the nucleus after reperfusion. Gene transfection with IkappaBalpha SR, but not with LacZ, markedly attenuated ischaemia/reperfusion injury, suppressing inducible nitric oxide synthase and nitrotyrosine expression in the liver. Moreover, no remarkable hepatocyte apoptosis was detected under IkappaBalpha SR overexpression. CONCLUSIONS: Adenoviral transfer of the IkappaBalpha SR gene in the liver ameliorates short term warm ischaemia/reperfusion injury, possibly through attenuation of hepatic macrophage activation. PMID- 15888795 TI - Ephrin-A1 expression contributes to the malignant characteristics of {alpha} fetoprotein producing hepatocellular carcinoma. AB - BACKGROUND AND AIMS: alpha-Fetoprotein (AFP), a tumour marker for hepatocellular carcinoma (HCC), is associated with poor prognosis. Using cDNA microarray analysis, we previously found that ephrin-A1, an angiogenic factor, is the most differentially overexpressed gene in AFP producing hepatoma cell lines. In the present study, we investigated the significance of ephrin-A1 expression in HCC. METHODS: We examined ephrin-A1 expression and its effect on cell proliferation and gene expression in five AFP producing hepatoma cell lines, three AFP negative hepatoma cell lines, and 20 human HCC specimens. RESULTS: Ephrin-A1 expression levels were lowest in normal liver tissue, elevated in cirrhotic tissue, and further elevated in HCC specimens. Ephrin-A1 expression was strongly correlated with AFP expression (r = 0.866). We showed that ephrin-A1 induced expression of AFP. This finding implicates ephrin-A1 in the mechanism of AFP induction in HCC. Ephrin-A1 promoted the proliferation of ephrin-A1 underexpressing HLE cells, and an ephrin-A1 antisense oligonucleotide inhibited the proliferation of ephrin-A1 overexpressing Huh7 cells. Thus ephrin-A1 affects hepatoma cell growth. cDNA microarray analysis showed that ephrin-A1 induced expression of genes related to the cell cycle (p21), angiogenesis (angiopoietin 1 and thrombospondin 1), and cell-cell interactions (Rho, integrin, and matrix metalloproteinases) in cultured hepatoma cells. These ephrin-A1 induced genes are also activated in HCC tissues that overexpress AFP. CONCLUSION: These findings suggest that the poor prognosis of patients with AFP producing HCC is partially caused by ephrin-A1 expression, which induces expression of genes related to tumour cell growth, angiogenesis, invasion, and metastasis. PMID- 15888796 TI - Hepatitis C virus (HCV) genotypes in 373 Italian children with HCV infection: changing distribution and correlation with clinical features and outcome. AB - BACKGROUND AND AIMS: Little is known of hepatitis C virus (HCV) genotypes in HCV infected children. This retrospective, multicentre study investigated genotype distribution and correlation with clinical features and outcome in a large series of Italian children. METHODS: Between 1990 and 2002, 373 HCV RNA positive children, consecutively recruited in 15 centres, were assayed for genotypes by a commercial line probe assay. RESULTS: The following genotype distribution pattern was recorded: genotype 1b = 41%; 1a = 20%; 2 = 17%; 3 = 14.5%; 4 = 5%; other = 2.5%. The prevalence of genotypes 1b and 2 decreased significantly (p<0.001) among children born from 1990 onwards compared with older children (46% v 70%) while the rate of genotypes 3 and 4 increased significantly (from 8% to 30%). Children infected with genotype 3 had the highest alanine aminotransferase levels and the highest rate of spontaneous viraemia clearance within the first three years of life (32% v 3% in children with genotype 1; p<0.001). Of 96 children enrolled in interferon trials during the survey, 22% definitely lost HCV RNA, including 57% of those with genotypes 2 and 3. CONCLUSION: HCV genotypes 1 and 2 are still prevalent among infected adolescents and young adults in Italy but rates of infection with genotypes 3 and 4 are rapidly increasing among children. These changes could modify the clinical pattern of hepatitis C in forthcoming years as children infected with genotype 3 have the best chance of spontaneous viraemia clearance early in life, and respond to interferon in a high proportion of cases. PMID- 15888797 TI - Peginterferon {alpha}-2b and ribavirin therapy in chronic hepatitis C genotype 4: impact of treatment duration and viral kinetics on sustained virological response. AB - BACKGROUND: The response rates and duration of peginterferon alpha (PEG-IFN alpha) and ribavirin combination therapy in chronic hepatitis C genotype 4, the prevalent genotype in the Middle East and Africa, are poorly documented. AIMS: To compare the efficacy and safety of 24, 36, or 48 weeks of PEG-IFN-alpha-2b and ribavirin therapy in chronic hepatitis C genotype 4. METHODS: In this prospective, randomised, double blind study, 287 patients with chronic hepatitis C genotype 4 were randomly assigned to PEG-IFN-alpha-2b (1.5 mug/kg) once weekly plus daily ribavirin (1000-1200 mg) for 24 weeks (group A, n = 95), 36 weeks (group B, n = 96), or 48 weeks (group C, n = 96) and followed for 48 weeks after completion of treatment. Early viral kinetics and histopathological evaluation of pre- and post treatment liver biopsies were performed. The primary end point was viral clearance 48 weeks after completion of treatment. RESULTS: Sustained virological response was achieved in 29%, 66%, and 69% of patients treated with PEG-IFN-alpha-2b and ribavirin for 24, 36, and 48 weeks, respectively, by intention to treat analysis. No statistically significant difference in sustained virological response rates was detected between 36 and 48 weeks of therapy (p = 0.3). Subjects with sustained virological response showed greater antiviral efficacy (epsilon) and rapid viral load decline from baseline to treatment week 4 compared with non-responders and improvement in liver histology. The incidence of adverse events was higher in the group treated for 48 weeks. CONCLUSION: PEG-IFN alpha-2b and ribavirin for 36 or 48 weeks was more effective in the treatment of chronic hepatitis C genotype 4 than treatment for 24 weeks. Thirty six week therapy was well tolerated and produced sustained virological and histological response rates similar to the 48 week regimen. PMID- 15888798 TI - Clinical relevance of proteinase activated receptors (pars) in the gut. PMID- 15888800 TI - Acute toxic gastric mucosal damage induced by Lugol's iodine spray during chromoendoscopy. PMID- 15888799 TI - Endoscopic therapy for Barrett's oesophagus. PMID- 15888801 TI - Erosions or not in GORD? The potential role of oesophageal cell proliferation. PMID- 15888802 TI - Gastric ulceration due to chronic mesenteric ischaemia treated by stenting of the inferior mesenteric artery. PMID- 15888803 TI - Role of IL-10 promoter haplotypes in Helicobacter pylori associated gastric inflammation. PMID- 15888804 TI - Diagnostic radiation exposure and cancer risk. PMID- 15888805 TI - Can gastro-oesophageal reflux be predicted while advancing the endoscope through the laryngeal area? PMID- 15888806 TI - Adalimumab use in pregnancy. PMID- 15888807 TI - CD40 antisense based strategy for inflammatory bowel disease: shutting down multiple cellular communication systems. PMID- 15888808 TI - Is stool DNA multitarget testing an unreliable strategy for colorectal cancer screening? PMID- 15888810 TI - The NIH Public Access Policy: CHEST's position and concerns regarding the responsibilities of the NIH principle investigator. PMID- 15888811 TI - Evaluating cardiorespiratory fitness after stroke: does the best provide less? PMID- 15888809 TI - Guidelines for the management of gastroenteropancreatic neuroendocrine (including carcinoid) tumours. PMID- 15888812 TI - Role of endothelin-1 in acute myocardial infarction. PMID- 15888813 TI - The interaction of vascular stiffness and cardiovascular events in women: insights from The Heart and Estrogen/Progestin Replacement Study. PMID- 15888814 TI - Severe pulmonary hypertension in COPD: is it a distinct disease? PMID- 15888815 TI - Not all that comes out is hot air. PMID- 15888816 TI - Risk of surgical lung biopsy in idiopathic interstitial pneumonias. PMID- 15888817 TI - Heartburn: another danger in the night? PMID- 15888818 TI - Finding haystacks full of needles: from Opus to Osler. PMID- 15888819 TI - Prognostic value of circulating levels of endothelin-1 in patients after acute myocardial infarction undergoing primary coronary angioplasty. AB - BACKGROUND: The link between increased circulating level of endothelin (ET)-1 and adverse clinical outcomes after acute myocardial infarction (AMI) has been established. Current studies demonstrate that reperfusion therapy by either thrombolysis or primary percutaneous coronary intervention (PCI) can salvage myocardium, improving survival of AMI patients. However, whether reperfusion therapy by primary PCI can prevent the adverse effect of ET-1 on clinical outcomes in patients after AMI remains unclear. Therefore, this study examined the predictive value of circulating ET-1 levels on 30-day outcomes in ST-segment elevated AMI treated with primary PCI. METHODS AND RESULTS: We conducted a prospective cohort study of 186 consecutive patients with ST-segment elevated AMI of onset < 12 h who underwent primary PCI. Blood samples for plasma concentration of ET-1 were collected in the catheterization laboratory following vascular puncture. Patients were classified into a high group (group 1, ET-1 level >or= 0.632 pg/mL, n = 93) and a low group (group 2, ET-1 level < 0.632 pg/mL, n = 93) according to the median value of ET-1 after AMI. Univariate analysis demonstrated that the 30-day composite major adverse clinical outcomes (MACO) [advanced Killip score >or= 3], severe congestive heart failure (CHF) [New York Heart Association functional class 4], and 30-day mortality were strongly associated with high ET-1 level (>or= 0.632 pg/mL; p < 0.0001), unsuccessful reperfusion (final Thrombolysis in Myocardial Infarction flow 15 apneas and hypopneas per hour of sleep during polysomnography or an oxygen desaturation index of > 10 events per hour during oximetry. The pattern of desaturation/resaturation during oximetry was also examined to distinguish obstructive events from central events. Using a 2% fall in pulse oximetric saturation as the criterion for oxygen desaturation, home oximetry had a 85% sensitivity and a 93% specificity (p < 0.001) for detecting an SRBD. However, the desaturation/resaturation pattern did not accurately distinguish between obstructive events and central events (eg, 100% sensitivity, 17% specificity for identifying CSA). The interpretation of the oximetry recording was highly consistent between scorers (p < 0.001). CONCLUSIONS: Overnight home oximetry is a sensitive and specific tool for identifying SRBDs in patients with CHF, but not for distinguishing between obstructive and central events in such patients. PMID- 15888822 TI - Effect of a centralized clinical pharmacy anticoagulation service on the outcomes of anticoagulation therapy. AB - CONTEXT: A growing body of reports has documented the ability of anticoagulation management services to help patients receiving warfarin therapy achieve better outcomes compared to the care provided by their personal physicians (ie, usual care). OBJECTIVE: To compare clinical outcomes associated with anticoagulation therapy provided by a clinical pharmacy anticoagulation service (CPAS) to usual care. DESIGN: Retrospective, observational cohort study, 6 months in duration. SETTING: Large nonprofit, group-model health maintenance organization. PATIENTS: A total of 6,645 patients receiving warfarin therapy were included in the final analyses (intervention group, 3,323 patients; control group, 3,322 patients). INTERVENTION: Anticoagulation therapy for patients in the intervention group was managed by a centralized, telephonic CPAS. Therapy for patients in the control group was managed in the usual manner by their personal physicians. MAIN OUTCOME MEASURES: The primary outcome was the occurrence of anticoagulation therapy related complications. A secondary outcome was the proportion of time spent in the target international normalized ratio (INR) range for each patient. Cox proportional hazards regression analyses were used to examine the risk of complications in relation to the study group. RESULTS: Patients in the CPAS were 39% less likely to experience an anticoagulation therapy-related complication than were patients in the control group (hazard ratio, 0.61; 95% confidence interval, 0.42 to 0.88). The number of patients needed to treat to prevent an anticoagulation therapy complication was 52. Additional analyses revealed that improved outcomes associated with CPAS were mediated largely through improved therapeutic INR control. Patients in the CPAS group spent 63.5% of study period days within their target INR range compared to 55.2% in the control group (p < 0.001). CONCLUSIONS: A centralized, telephonic, pharmacist-managed anticoagulation monitoring service reduced the risk of anticoagulation therapy related complications compared to that with usual care. The cumulative evidence supporting the superior care associated with implementing a pharmacist-managed anticoagulation monitoring service was sufficient to recommend widespread implementation. PMID- 15888823 TI - Thalassemia heart disease: a comparative evaluation of thalassemia major and thalassemia intermedia. AB - BACKGROUND: Heart disease represents the main determinant of survival in beta thalassemia, but its particular features in the two clinical forms of the disease, thalassemia major (TM) and thalassemia intermedia (TI), are not completely clarified. METHODS: We compared clinical and echocardiographic global parameters in 131 TM patients who received regular chelation transfusions and were highly compliant with treatment (mean age, 28 +/- 6 years [+/- SD]), and 74 age-matched, TI patients who did not receive chelation transfusions. RESULTS: Congestive heart failure was encountered in five patients with TM (3.8%; age range, 25 to 29 years) and in two patients with TI (2.7%; age range, 37 to 40 years). Systolic left ventricular (LV) dysfunction (ejection fraction < 55% or shortening fraction < 35%) was only encountered in patients with TM (8.4%). Considerable pulmonary hypertension (systolic tricuspid gradient > 35 mm Hg) was only present in TI (23.0%). In the remaining patients without evident heart disease, cardiac dimensions, LV mass, LV shortening and ejection fractions, and cardiac output were significantly higher in patients with TI. LV afterload was higher in patients with TM. LV diastolic early transmitral diastolic peak flow velocity (E)/late transmitral diastolic peak flow velocity (A) ratio was also higher in TM. Systolic and mean pulmonary artery pressures and total pulmonary resistance were higher in both young and old TI patients. CONCLUSION: Regular lifelong transfusion and chelation therapy in TM prevented premature heart disease and pulmonary hypertension, although LV dysfunction still occurred and led to heart failure. The absence of regular therapy in TI, in contrast, preserved systolic LV function but allowed pulmonary hypertension development, which also led to heart failure, starting within the fourth decade of life, a decade later compared to TM. PMID- 15888824 TI - Pulmonary hemodynamics in advanced COPD candidates for lung volume reduction surgery or lung transplantation. AB - STUDY OBJECTIVES: To assess the pulmonary hemodynamic characteristics in COPD candidates for lung volume reduction surgery (LVRS) or lung transplantation (LT). DESIGN: Retrospective study. SETTING: One center in France. PATIENTS: Two hundred fifteen patients with severe COPD who underwent right-heart catheterization before LVRS or LT. RESULTS: Mean age was 54.6 years. Pulmonary function test results were as follows: FEV(1), 24.3% predicted; total lung capacity, 128.3% predicted; residual volume, 259.7% predicted. Mean pulmonary artery pressure (PAPm) was 26.9 mm Hg. Pulmonary hypertension (PAPm > 25 mm Hg) was present in 50.2% and was moderate (PAPm, 35 to 45 mm Hg) or severe (PAPm > 45 mm Hg) in 9.8% and in 3.7% of patients, respectively. Cardiac index was low normal. PAPm was related to Pao(2) and alveolar-arterial oxygen gradient in multivariate analysis. Cluster analysis identified a subgroup of atypical patients (n = 16, 7.4%) characterized by moderate impairment of the pulmonary mechanics (mean FEV(1), 48.5%) contrasting with high level of pulmonary artery pressure (PAPm, 39.8 mm Hg), and severe hypoxemia (mean Pao(2), 46.2 mm Hg). CONCLUSION: While pulmonary hypertension is observed in half of the COPD patients with advanced disease, moderate-to-severe pulmonary hypertension is not a rare event in these patients. We individualized a subgroup of patients presenting with a predominant vascular disease that could potentially benefit from vasodilators. PMID- 15888825 TI - Glutathione S-transferase P1 and lung function in patients with alpha1 antitrypsin deficiency and COPD. AB - BACKGROUND: The glutathione S-transferase P1 (GSTP1) gene is involved in detoxification of electrophilic substances of tobacco smoke. A polymorphism at nucleotide 315 of this gene alters its enzymatic activity. OBJECTIVE: We analyzed the association between the variability in the GSTP1 gene and impairment in lung function in smokers with and without alpha(1)-antitrypsin (AAT) deficiency and COPD. POPULATION AND METHOD: The study population consisted of 99 patients with smoking-related COPD and 69 patients with AAT deficiency; 198 healthy volunteers provided the frequency of the different polymorphisms in the general population. GSTP1 genotyping was performed by a real-time polymerase chain reaction amplification assay. RESULTS: The frequency (0.28) of the 105Val polymorphism was identical in COPD patients and the general population. However, the frequency was significantly increased (0.44) in patients with AAT deficiency (odds ratio [OR], 2.09; 95% confidence interval [CI], 1.17 to 3.72 compared to control subjects; and OR, 2.41; 95% CI, 1.27 to 4.59 compared to COPD). FEV(1) percentage of predicted was significantly impaired in AAT-deficient carriers of 105Val. This effect was not observed in COPD patients. CONCLUSIONS: These findings suggest that the frequency of the GSTP1 105Val polymorphism is increased in patients with AAT deficiency. Globally, GSTP1 genotypes, age, and tobacco smoking explained 41% of total FEV(1) percentage of predicted variability in patients with AAT deficiency. The modulatory role of GSTP1 in lung disease has only been observed in smokers lacking AAT. PMID- 15888826 TI - Ten-year cumulative incidence of COPD and risk factors for incident disease in a symptomatic cohort. AB - STUDY OBJECTIVES: To determine the 10-year cumulative incidence of COPD in a cohort of subjects with respiratory symptoms (Global Initiative for Chronic Obstructive Lung Disease [GOLD] stage 0) using the British Thoracic Society (BTS) and GOLD spirometric criteria. Furthermore, we sought to evaluate risk and gender factors for incident COPD. DESIGN AND SETTING: A postal questionnaire was administered in 1986 to all 6,610 subjects in eight areas of northern Sweden who had been born in 1919 to 1920 (group 1), 1934 to 1935 (group 2), and 1949 to 1950 (group 3). The response rate was 86%. All of the subjects reporting respiratory symptoms were invited to participate in a structured interview and pulmonary function test (PFT), and 1,506 (91%) participated. In 1996, 90% could be traced for follow-up, of whom 1,165 (86%) of the invited subjects participated and 1,109 subjects (534 women) were able to perform technically adequate PFTs in both 1986 and 1996. RESULTS: The 10-year cumulative incidence of COPD was estimated at 8.2% (using BTS criteria) and 13.5% (using GOLD criteria). Significant risk factors for incident COPD (using BTS and GOLD criteria) in a multivariate analysis were higher age (group 1 odds ratio [OR]: BTS criteria, 3.49; GOLD criteria, 3.37; group 2 OR: BTS criteria, 4.50; GOLD criteria, 5.70) and smoking (OR: BTS criteria, 5.37; GOLD criteria, 4.56), but not gender or heredity. Respiratory symptoms were significantly associated with incident COPD when added to the same model. In analogous analyses that were conducted separately for men and women, smoking yielded an OR of 8.52 among women (95% confidence interval [CI], 3.43 to 21.2) compared with 3.14 among men (95% CI, 1.26 to 7.84). The symptoms cough, sputum production, and chronic productive cough reached statistical significance in women, while dyspnea and wheeze did so in men. CONCLUSION: In this cohort, the 10-year cumulative incidence of COPD was 8.2% (using BTS criteria) and 13.5% (using GOLD criteria). Increasing age, smoking, and bronchitic symptoms, but not gender, were risk factors for incident COPD. GOLD stage 0 therefore appears to identify subjects who are at risk of COPD, but men and women presented different risk profiles. PMID- 15888827 TI - Leukotriene B4 in exhaled breath condensate and sputum supernatant in patients with COPD and asthma. AB - STUDY OBJECTIVES: Some patients with COPD present with significant reversibility of airflow limitation after receiving bronchodilation therapy. Leukotriene B(4) (LTB(4)) has been implicated in the pathophysiology of both COPD and asthma. We tested the hypothesis that COPD patients with airflow reversibility and asthmatic patients who smoke might have similar levels of LTB(4) in exhaled breath condensate (EBC) and sputum supernatant. The repeatability and stability of LTB(4) measurements were additionally studied. DESIGN: Prospective, cross sectional study. PATIENTS OR PARTICIPANTS: We studied 30 patients with COPD (15 smokers [FEV(1), 56% predicted; SD, 6% predicted]; 15 patients with significant reversibility in airway obstruction after bronchodilation [FEV(1), 14% predicted; SD, 2% predicted]). Fifteen asthmatic patients who smoked, with similar FEV(1) and reversibility were also studied. Ten healthy smokers served as control subjects. SETTING: A hospital research laboratory. INTERVENTIONS: Spirometry and reversibility testing were performed on the first visit. On the following day, EBC was collected for the measurement of LTB(4), and induced sputum was collected for differential cell counts and LTB(4) measurement in the sputum supernatant. MEASUREMENTS AND RESULTS: LTB(4) levels in EBC [mean (SD)] were increased in COPD patients (mean, 86.7 pg/mL; SD, 19 pg/mL) and asthmatic patients (mean, 97.5 pg/mL; SD, 15 pg/mL) compared to control subjects (mean, 32.3 pg/mL; SD, 10 pg/mL; p < 0.0001 for both groups). COPD patients with airflow reversibility (mean, 99.8 pg/mL; SD, 12 pg/mL) had values similar to those of asthmatic patients (mean, 97.5 pg/mL; SD, 15 pg/mL; p = 0.2) and higher than those of COPD patients without airflow reversibility (mean, 73.7 pg/mL; SD, 17 pg/mL; p = 0.002). Similar results were observed in the sputum supernatant. Measurements of LTB(4) in EBC and sputum were repeatable on two consecutive days, but measurements in the frozen samples of EBC and sputum were not stable after 3 weeks. CONCLUSIONS: Patients with asthma and reversible COPD presented with higher LTB(4) values compared to patients with nonreversible COPD and healthy smokers. This difference may be mainly attributed to the presence of reversibility in airway obstruction, probably as part of a common underlying inflammatory process. PMID- 15888828 TI - FEV1/FEV6 and FEV6 as an alternative for FEV1/FVC and FVC in the spirometric detection of airway obstruction and restriction. AB - STUDY OBJECTIVES: To evaluate the use of the FEV(1)/forced expiratory volume at 6 s of exhalation (FEV(6)) ratio and FEV(6) as an alternative for FEV(1)/FVC and FVC in the detection of airway obstruction and lung restriction, respectively. SETTING: Pulmonary function laboratory of the Academic Hospital of the Free University of Brussels. PARTICIPANTS: A total of 11,676 spirometric examinations were analyzed on subjects with the following characteristics: white race; 20 to 80 years of age; 7,010 men and 4,666 women; and able to exhale for at least 6 s. METHODS: Published reference equations were used to determine lower limits of normal (LLN) for FEV(6), FVC, FEV(1)/FEV(6), and FEV(1)/FVC. We considered a subject to have obstruction if FEV(1)/FVC was below its LLN. A restrictive spirometric pattern was defined as FVC below its LLN, in the absence of obstruction. From these data, sensitivity and specificity of FEV(1)/FEV(6) and FEV(6) were calculated. RESULTS: For the spirometric diagnosis of airway obstruction, FEV(1)/FEV(6) sensitivity was 94.0% and specificity was 93.1%; the positive predictive value (PPV) and negative predictive value (NPV) were 89.8% and 96.0%, respectively. The prevalence of obstruction in the entire study population was 39.5%. For the spirometric detection of a restrictive pattern, FEV(6) sensitivity was 83.2% and specificity was 99.6%; the PPVs and NPVs were 97.4% and 96.9%, respectively. The prevalence of a restrictive pattern was 15.7%. Similar results were obtained for male and female subjects. When diagnostic interpretation differed between the two indexes, measured values were close to the LLN. CONCLUSIONS: The FEV(1)/FEV(6) ratio can be used as a valid alternative for FEV(1)/FVC in the diagnosis of airway obstruction, especially for screening purposes in high-risk populations for COPD in primary care. In addition, FEV(6) is an acceptable surrogate for FVC in the detection of a spirometric restrictive pattern. Using FEV(6) instead of FVC has the advantage that the end of a spirometric examination is more explicitly defined and is easier to achieve. PMID- 15888829 TI - Effect of pet removal on pet allergic asthma. AB - BACKGROUND: Allergen avoidance has been recommended in the management of allergic asthma. Very few studies have assessed the effect of pet removal on pet allergic asthma. OBJECTIVE: We examined the effect of pet removal from homes on pulmonary function testing, airway hyperresponsiveness, and medication use. DESIGN: Prospective, nonrandomized, nonblinded observational study. PATIENTS AND METHODS: Subjects included 20 symptomatic patients with newly diagnosed pet allergic asthma who were keeping domestic animals, including hamsters, cats, dogs, and ferrets, and were sensitized to the animals. They were treated with inhaled corticosteroids or other medications according to recommendations by level of severity of the Global Initiative for Asthma. Methacholine inhalation tests were performed regularly before and after starting medication. Clinical features were compared between the patients who gave away their pets according to recommendations by the clinician (removal group) and the patients who refused to give away their pets (keeping group). RESULTS: There were 10 patients in both the removal group and the keeping group. After >or= 1 year of follow-up with or without pet removal, a 5.9-fold increase in the provocative concentration of methacholine causing a 20% fall in FEV(1) was observed in the removal group compared with a 2.3-fold increase in the keeping group (p = 0.04). There were no significant differences in the changes in FEV(1) and peak flow variability. Finally, no patient received inhaled corticosteroids in the removal group, whereas all but one of the patients needed beclomethasone dipropionate (mean dose, 600 mug/d) in the keeping group. CONCLUSION: This study indicates that removal of pets from homes reduces airway responsiveness in patients with pet allergic asthma more than optimal pharmacotherapy alone, thereby enabling a decrease in inhaled corticosteroid doses. PMID- 15888830 TI - Bronchoprotective effects of single doses of salmeterol combined with montelukast in thermally induced bronchospasm. AB - STUDY OBJECTIVES: Salmeterol (S) and montelukast (M) individually inhibit the obstructive consequences of thermal stimuli such as exercise and hyperventilation (HV), but there is no information on whether these drugs can interact positively. DESIGN: Randomized trial. SETTING: University teaching hospital. PARTICIPANTS: Atopic asthmatic patients with sensitivity to thermal provocations. INTERVENTIONS: Eleven asthmatic patients generated stimulus-response curves to isocapnic HV while breathing frigid air without any interventions and then after pretreatment with 42 mug of S, 10 mg of M, and the combination. The order of testing was randomly determined. MEASUREMENTS AND RESULTS: Minute ventilation (Ve) was increased in 20-L increments until FEV(1) fell >or= 15%. Measurements were obtained before and 1 h after drug administration, and then again 5 min after each bout of HV. In the nonintervention trial, the provocation commenced after the patients presented to the laboratory. In the control challenge, the mean (+/- SEM) FEV(1) decreased 24.6 +/- 1.7% from baseline. S and M both increased the mean prechallenge FEV(1) significantly (S, 10.4 +/- 1.7% [p < 0.01]; M, 4.1 +/- 1.3% [p = 0.02]; S + M, p = 0.01). The combination of S + M produced greater bronchodilatation (mean improvement, 12.4 +/- 2.3%) than M alone (p = 0.004), but not greater than S alone (p = 0.80). Both drugs blunted the obstructive response similarly (protection: M, 34.6 +/- 15.1%; S, 60 +/- 8.7%; p = 0.13). The benefits added arithmetically with the combined regimen (protection with S + M, 84.9 +/- 5.5%; p = 0.01 vs S alone; p = 0.003 vs M alone). CONCLUSION: These data indicate that the concurrent administration of single standard doses of S and M appears to provide greater protection against thermal stimuli than does either drug alone. Further experimentation will be required to ascertain whether the combination will provide additional clinical benefits to patients over those of the single agents. PMID- 15888831 TI - Asthma exacerbations in North American adults: who are the "frequent fliers" in the emergency department? AB - OBJECTIVE: To characterize adult asthma patients according to frequency of emergency department (ED) visits in the past year. DESIGN: Adults presenting with acute asthma to 83 US EDs underwent structured interviews in the ED and by telephone 2 weeks later. RESULTS: The 3,151 enrolled patients were classified into four groups: those reporting no ED visits in the past year (27%), one to two visits (27%), three to five visits (25%), and six or more visits (21%). The number of ED visits (NEDV) was associated with older age, nonwhite race, lower socioeconomic status, and several markers of chronic asthma severity (all p < 0.001). NEDV was strongly associated with Medicaid insurance (17% among those with no visits, 22% with one to two visits, 30% with three to five visits, 39% with six or more visits; p < 0.001). NEDV was unrelated to gender or having a primary care provider (PCP). In a multivariate model, independent predictors of high ED use (six or more visits a year) were nonwhite race, Medicaid, other public, and no insurance, and markers of chronic asthma severity. Patients with six or more ED visits accounted for 67% of all prior ED visits in the past year. CONCLUSIONS: High NEDV is associated with characteristics that may help with identification of "frequent fliers" in the ED. A better understanding of these characteristics may advance ongoing efforts to decrease asthma health-care disparities, including differential access to primary asthma care. National guidelines recommend specific ED treatments then referral to a PCP. Although longitudinal care is surely important, attempts to reduce frequent ED asthma visits may be better directed toward more specific preventive and educational needs. PMID- 15888832 TI - Muscular strength and function in patients with cystic fibrosis. AB - BACKGROUND: For 20 years, physical activity has been an important component in the treatment of cystic fibrosis (CF) patients in Sweden. Data concerning physical performance in terms of muscular strength in these patients are limited OBJECTIVE: To compare muscular strength and function in patients with CF with those aspects in a healthy control group (CG). DESIGN: Thirty-three patients with CF (16 women) aged 16 to 35 years and 20 healthy individuals matched for age and gender were included in the study. All participants had undertaken regular physical training two to three times per week. The following tests were performed: vertical jumping ability; hand-grip strength; abdominal strength; arm/shoulder strength; quadriceps muscle strength; and a functional test of leg muscle endurance. RESULTS: Patients with CF showed decreased muscle strength and function compared to control subjects (women: maximal hand-grip strength in the right [p = 0.02] and the left hand [p = 0.001]; sustained hand-grip strength in the left hand [p = 0.002]; and in leg muscle endurance [p = 0.02]; men: the number of sit-ups performed within 30 s [p = 0.03]; and left leg isokinetic quadriceps strength at 180 degrees per second [p = 0.02]). The differences were not related to pancreatic or pulmonary function. There was no significant difference between the CF group and the CG in any other test results. CONCLUSIONS: Our study showed few differences in muscular performance between patients with CF and healthy control subjects. Both groups had regular moderate to-high activity levels. Further studies are needed to evaluate whether the small but significant differences might be related to metabolic abnormalities in skeletal muscles in CF patients. PMID- 15888833 TI - Cystic fibrosis patients have poor sleep quality despite normal sleep latency and efficiency. AB - STUDY OBJECTIVES: Cystic fibrosis (CF) patients may be predisposed to poor sleep quality due to upper and lower airway abnormalities and impaired gas exchange. Previous sleep investigations of CF patients using single-night polysomnography have reported conflicting results. We hypothesized that sampling sleep for a prolonged period in a patient's normal environment may give a more representative assessment of sleep quality than a single-night polysomnogram, and that impaired sleep quality would correlate with pulmonary disease severity and self-assessed sleep quality. DESIGN: Using wrist actigraphy, we measured sleep quality in clinically stable CF patients and age-matched control subjects. In addition, each CF patient and control subject completed the following three questionnaires: the Epworth sleepiness scale; the Pittsburgh sleep quality index (PSQI); and the Medical Outcomes Study 36-item short form. RESULTS: Twenty CF patients and control subjects were enrolled in the study, and were well-matched for age, sex, and body mass index. The mean (+/- SD) FEV(1) for CF patients was 61.0 +/- 20.1% predicted. CF patients and control subjects had similar sleep duration, sleep latency, and sleep efficiency. However, CF patients had higher PSQI scores (6.45 vs 4.55, respectively; p = .04), a higher fragmentation index (FI) [31.72 vs 18.02, respectively; p < 0.001], and less immobile time (88.87 vs 91.89, respectively; p = 0.02). There was a significant correlation of FI with FEV(1) and PSQI scores. CONCLUSIONS: Stable CF patients have disrupted sleep, and sleep disruption may in part be related to the severity of pulmonary disease. In addition, the PSQI may be useful in detecting CF patients with poor sleep quality. PMID- 15888834 TI - Outcomes and safety of surgical lung biopsy for interstitial lung disease. AB - STUDY OBJECTIVES: To determine the safety of surgical lung biopsy (SLB) in patients with interstitial lung disease (ILD), and specifically in those with idiopathic pulmonary fibrosis (IPF). DESIGN: Retrospective cohort. SETTING: Tertiary care university-affiliated military medical center. PATIENTS: Individuals undergoing SLB for suspected ILD. MEASUREMENTS AND RESULTS: We examined outcomes for subjects with a clinical diagnosis of ILD who had been designated to undergo SLB. Mortality (assessed at 30 and 90 days) following SLB represented the primary end point. Morbidity resulting from complications from SLB served as a secondary end point. The cohort included 83 patients (mean [+/- SD] age, 57.3 +/- 14.2 years; men, 57.8%). IPF was eventually diagnosed in slightly more than half of the subjects. Overall, 30-day and 90-day mortality rates were low (4.8% and 6.0%, respectively). Subjects with IPF did well with SLB (30-day mortality rate, 7.1%) and did not face a higher risk of either death or complications relative to individuals with non-IPF forms of ILD. The only predictors of perioperative mortality were either the need for mechanical ventilation (MV) at the time of SLB or being immunosuppressed prior to undergoing SLB. Excluding persons who met either criterion yielded an overall 90-day post SLB mortality rate of 1.5% in persons with IPF. Approximately 40% of patients in whom IPF was eventually diagnosed were initially thought to have another form of ILD. CONCLUSIONS: Persons with IPF tolerate SLB well. Requiring MV or being immunosuppressed is associated with an increased risk for death following SLB. Safety concerns should not preclude referral for SLB in patients who are clinically suspected of having IPF. PMID- 15888835 TI - CT scan findings in chronic thromboembolic pulmonary hypertension: predictors of hemodynamic improvement after pulmonary thromboendarterectomy. AB - STUDY OBJECTIVES: The aim was to correlate CT scan findings with hemodynamic measurements in patients who had undergone pulmonary thromboendarterectomy (PTE) and to evaluate whether CT scan findings can help to predict surgical outcome. PATIENTS AND METHOD: Sixty patients who underwent PTE and preoperative helical CT scanning were included. Preoperative and postoperative hemodynamics were correlated with preoperative CT imaging features. RESULTS: The diameter of the main pulmonary artery (PA) and the ratio of the PA and the diameter of the ascending aorta correlated with preoperative mean pulmonary artery pressure (PAP) [r = 0.42; p < 0.001; and r = 0.48; p < 0.0001, respectively]. There was a significant correlation of subpleural densities with preoperative pulmonary vascular resistance (PVR) [r = 0.44; p < 0.001] and of the number of abnormal perfused lobes with preoperative PAP (r = 0.66; p < 0.0001) and PVR (r = 0.76; p < 0.0001). Postoperative PVR correlated negatively with the presence and extent of central thrombi (r = -0.36; p = 0.007) and dilated bronchial arteries (p = 0.03) seen on preoperative CT scans. Sixty percent of patients (3 of 5 patients) without visible central thromboembolic material on CT scans had an inadequate hemodynamic improvement in contrast to 4% of patients (2 of 51 patients) with central thrombi (p = 0.003). Preoperative PVR (r = 0.31; p = 0.018) and the extent of abnormal lung perfusion (r = 0.37; p = 0.007) and of subpleural densities (r = 0.32; p = 0.017) were positively correlated with postoperative PVR. CONCLUSIONS: In patients with thromboembolic pulmonary hypertension, CT scan findings can help to predict hemodynamic improvement after PTE. The absence of central thrombi is a significant risk factor for inadequate hemodynamic improvement. PMID- 15888836 TI - Variation in training for interventional pulmonary procedures among US pulmonary/critical care fellowships: a survey of fellowship directors. AB - STUDY OBJECTIVES: The American College of Chest Physicians has published guidelines recommending minimum competency requirements for 17 interventional pulmonary procedures. Our aim was to assess what procedures are offered to fellows in US pulmonary/critical care fellowships and to determine whether the recommended competency numbers are being met. METHODS: Surveys were mailed to 122 pulmonary/critical care fellowship directors in the United States, and fellowship demographics, the types of procedures offered, and the average number of procedures performed were recorded. The presence of a dedicated interventional pulmonologist (IP) was ascertained, and procedural offerings and volume were compared with programs that did not have an IP. RESULTS: The response rate of the survey was 77%. There was wide variation in the procedures offered by different programs. The presence of an IP was associated with an increased likelihood of advanced procedural training in brachytherapy (p < 0.05), electrocautery/argon plasma coagulation (p < 0.001), stents (p < 0.001), laser therapy (p < 0.01), rigid bronchoscopy (p < 0.001), and cryotherapy (p < 0.05). For only 3 of the 17 procedures did > 50% of the programs reach the targeted numbers to obtain competency. CONCLUSIONS: There is a large variation in the spectrum of pulmonary procedures offered to trainees. Programs with a dedicated IP are more likely to offer training in advanced therapeutic procedures. When interventional procedures are offered by fellowships, < 30% of programs meet the competency recommendations. These findings have implications for training, delivery of care, and research. An extra year of fellowship in interventional pulmonology might be desirable if one is to reach the desired competency numbers. An alternative to reaching the recommended numbers for select procedures would be to consider regionalizing care at centers that perform many procedures. Finally, to provide justification for the current competency recommendations, clinical outcomes should be correlated with physicians' procedural volume, as has been done in other subspecialties. PMID- 15888837 TI - Preoperative aspirin administration improves oxygenation in patients undergoing coronary artery bypass grafting. AB - OBJECTIVES: Release of thromboxane (Tx) A(2) by platelets may be one of multiple factors that are responsible for lung injury after cardiopulmonary bypass, leading to pulmonary vasoconstriction and impaired oxygenation. In experimental models, the inhibition of Tx receptor or its production improved lung function. The use of aspirin, which is used widely in the treatment of ischemic heart disease because of its antiplatelet activity, is usually discontinued a week before the patient undergoes the operation to restore normal platelet hemostatic function. The purpose of this study was to determine the relationship between the time of cessation of aspirin before coronary artery bypass surgery, and postoperative oxygenation and bleeding. DESIGN: A prospective clinical study comparing the effect of aspirin on postoperative oxygenation in patients who had been treated or had not been treated with aspirin. SETTING: Tx levels in the pericardial fluid, oxygenation, and bleeding were compared between the two groups. PATIENTS: Thirty-two patients with coronary artery disease who were undergoing coronary artery bypass grafting. Fourteen of these patients received aspirin until the day of the operation, whereas 18 patients stopped receiving aspirin at least 1 week before undergoing the operation. MAIN RESULTS: Mean (+/- SD) Tx levels in the pericardial fluid were significantly lower in the aspirin group (117 +/- 47 pg/mL) compared to those in the control group (1,306 +/- 2,048 pg/mL; p = 0.02). The duration of ventilation after the operation was significantly longer in the nonaspirin group (9.6 +/- 5.6 h vs 3.8 +/- 1.4 h, respectively; p = 0.0004). Po(2) reached a higher level while patients breathed 100% O(2) in the aspirin group (235 +/- 54 mm Hg vs 176 +/- 27 mm Hg, respectively; p = 0.001). The mean amount of bleeding during the first 24 h after surgery was increased in the aspirin group (710 +/- 202 mL) compared with the nonaspirin group (539 +/- 143 mL; p = 0.01), but these patients did not require more transfusions. CONCLUSIONS: The administration of aspirin until the operation may improve oxygenation with only a slight increase in bleeding. This improvement is probably mediated by antiplatelet activity and Tx inhibition by aspirin. PMID- 15888838 TI - Clinical gestalt and the diagnosis of pulmonary embolism: does experience matter? AB - STUDY OBJECTIVES: We sought to determine whether the accuracy of pretest assessment of the likelihood of pulmonary embolism (PE) was related to physician experience. We compared the accuracy of the subjective pretest probability assessment made by senior physicians (postgraduate year [PGY]-4+) to that of interns (PGY-1) and residents (PGY-2 and PGY-3) working in the emergency department of a large teaching hospital. DESIGN: Prospective observational study. SETTING: Urban, academic emergency department with an annual census of 48,000 patient visits. PATIENTS: Five hundred eighty-three adults evaluated for PE in the emergency department. INTERVENTIONS: Eligible patients had at least one diagnostic test ordered to workup PE. The physician treating the patient was asked whether he or she considered PE the most-likely diagnosis or whether an alternative diagnosis was most likely. This result was compared to the ultimate diagnosis. Physician experience was categorized by the number of years of training since medical school graduation. MEASUREMENTS AND RESULTS: There was a trend toward increasing accuracy with increasing experience, demonstrated by the frequency of true-positive assessments (17% vs 20% vs 25%), true-negative assessments (89% vs 94% vs 96%), and likelihood ratio (1.49 vs 2.34 vs 3.33), respectively. CONCLUSIONS: Accurate determination of the pretest probability of PE appears to increase with clinical experience. However, the difference in accuracy between inexperienced and experienced physicians is not sufficiently large to distinguish between the two when determining whether clinical gestalt or a clinical prediction rule should be used to determine the pretest probability of PE. PMID- 15888839 TI - Bed rest or ambulation in the initial treatment of patients with acute deep vein thrombosis or pulmonary embolism: findings from the RIETE registry. AB - BACKGROUND: Traditionally, many patients with acute deep vein thrombosis (DVT) are treated not only by anticoagulation therapy but additionally by strict bed rest, which is aimed at reducing the risk of pulmonary embolism (PE) events. However, this risk has not been subjected to empirical verification. PATIENTS AND METHODS: The Registro Informatizado de la Enfermedad TromboEmbolica is a Spanish registry of consecutively enrolled patients with objectively confirmed, symptomatic acute DVT or PE. In this analysis, the clinical characteristics, details of anticoagulant therapy, and clinical outcomes of enrolled patients with and without strict bed rest prescribed during the first 15 days were compared. Patients in whom ambulation was not possible were not included in this analysis. RESULTS: A total of 2,650 patients entered the study (DVT, 2,038 patients; PE, 612 patients). Of these patients, 1,050 DVT patients (52%) and 385 PE patients (63%) were prescribed strict bed rest. New events of symptomatic, objectively confirmed PE developed during the 15-day study period in 11 patients with DVT (0.5%) and 4 patients with PE (0.7%). Five of these 15 patients (33%) died as a result of their PE. Age < 65 years (odds ratio [OR], 3.1; 95% confidence interval [CI], 0.98 to 11) and cancer (OR, 3.0; 95% CI, 0.98 to 9.1) were associated with an increased rate of new PEs. There were not significant differences between bedridden and ambulant patients in terms of new PE events, fatal PE, or bleeding complications. CONCLUSIONS: Our findings confirm those from previous reports suggesting that bed rest has no influence on the risk of developing PE among patients with acute DVT of the lower limbs. In addition, our findings show for the first time the lack of influence of bed rest even in patients presenting with acute submassive PE. PMID- 15888840 TI - End-tidal PCO2 abnormality and exercise limitation in patients with primary pulmonary hypertension. AB - OBJECTIVES: Primary pulmonary hypertension (PPH) is a pulmonary vasculopathy resulting in exercise intolerance, usually due to dyspnea. We hypothesized that ventilation is increased during exercise in PPH relative to normal because the ventilated lung is underperfused, cardiac output increase is restricted, and arterial hypoxemia may develop. Our aim was to determine the size of the reduction in end-tidal Pco(2) (Petco(2)) as a reflection of the abnormality in ventilatory efficiency and ventilatory drive in PPH patients. METHODS: We performed cardiopulmonary exercise testing (CPET) in 52 PPH patients. All had hemodynamic measurements to confirm the diagnosis of PPH. A subgroup of 29 patients who underwent right-heart catheterization within 50 days of CPET were studied to compare their CPET responses to resting hemodynamics. Nine healthy volunteers matched for age and gender served as CPET control subjects. RESULTS: In PPH patients, the percentage of predicted peak oxygen uptake (Vo(2)) correlated significantly with mean pulmonary artery pressure (mPAP) [r = - 0.59, p = 0.0007, n = 29]. Petco(2) values at rest, anaerobic threshold (AT), and peak Vo(2) were proportionately reduced as percentage of predicted peak Vo(2) decreased (r = 0.66 to 0.72, p < 0.0001, n = 52). Petco(2) values at rest, AT, and peak Vo(2) were also reduced as mPAP increased (r = - 0.51 to - 0.53, p < 0.005, n = 29). In contrast to normal subjects in whom Petco(2) increased from rest to AT, Petco(2) decreased in PPH patients, except for two patients with mild PPH in whom there was no change. Also, Petco(2) increased rather than decreased further at the start of recovery, in contrast to normal. Although usually normal at rest, oxyhemoglobin saturation decreased during exercise in most PPH patients. CONCLUSIONS: In patients with PPH, Petco(2) at rest and exercise is significantly reduced in proportion to physiologic disease severity. The range of values is unusually low. Furthermore, the directional changes of Petco(2) during exercise and early recovery are in the opposite direction of normal. PMID- 15888841 TI - Hemodynamic effects of sildenafil in patients with congestive heart failure and pulmonary hypertension: combined administration with inhaled nitric oxide. AB - STUDY OBJECTIVES: In patients with pulmonary hypertension (PH) secondary to congestive heart failure, inhaled nitric oxide (NO) increases pulmonary vascular smooth-muscle intracellular cyclic guanosine monophosphate (cGMP) concentration, thereby decreasing pulmonary vascular resistance (PVR) and increasing cardiac index (CI). However, these beneficial effects of inhaled NO are limited in magnitude and duration, at least in part due to cGMP hydrolysis by the type 5 isoform of phosphodiesterase (PDE5). The goal of this study was to determine the acute pulmonary and systemic hemodynamic effects of the selective PDE5 inhibitor, sildenafil, administered alone or in combination with inhaled NO in patients with congestive heart failure and PH. DESIGN: Single center, case series, pharmacohemodynamic study. SETTING: Cardiac catheterization laboratory of a tertiary care academic teaching hospital. PATIENTS: We studied 11 patients with left ventricular systolic dysfunction due to coronary artery disease or idiopathic dilated cardiomyopathy who had PH. INTERVENTIONS: We administered oral sildenafil (50 mg), inhaled NO (80 ppm), and the combination of sildenafil and inhaled NO during right-heart and micromanometer left-heart catheterization. MEASUREMENTS AND RESULTS: Sildenafil administered alone decreased mean pulmonary artery pressure by 12 +/- 5%, PVR by 12 +/- 5%, systemic vascular resistance (SVR) by 13 +/- 6%, and pulmonary capillary wedge pressure by 12 +/- 7%, and increased CI by 14 +/- 5% (all p < 0.05) [+/- SEM]. The combination of inhaled NO and sildenafil decreased PVR by 50 +/- 4%, decreased SVR by 24 +/- 3%, and increased CI by 30 +/- 4% (all p < 0.01). These effects were greater than those observed with either agent alone (p < 0.05). In addition, sildenafil prolonged the pulmonary vasodilator effect of inhaled NO. Administration of sildenafil alone or in combination with inhaled NO did not change systemic arterial pressure or indexes of myocardial systolic or diastolic function. CONCLUSIONS: PDE5 inhibition with sildenafil improves cardiac output by balanced pulmonary and systemic vasodilation, and augments and prolongs the hemodynamic effects of inhaled NO in patients with chronic congestive heart failure and PH. PMID- 15888842 TI - Silent upper airway resistance syndrome: prevalence in a mixed military population. AB - STUDY OBJECTIVES: The upper airway resistance syndrome (UARS) is a recently described form of sleep-disordered breathing in which transient increases in upper airway resistance result in repetitive EEG arousals. UARS is not associated with apnea or diminished airflow, although snoring and excessive daytime somnolence (EDS) are common. This report describes a subset of patients with UARS diagnosed by polysomnography who do not manifest snoring, which we define as silent upper airway resistance syndrome (SUARS). DESIGN: A retrospective review of all polysomnographies performed at our sleep disorders center during 2000. SETTING: Sleep disorders center of a large, academic, military hospital. PATIENTS: Our center serves military personnel, military retirees, and their dependent families. INTERVENTIONS: Esophageal manometry during polysomnography was routinely performed on patients with hypersomnolence (Epworth sleepiness scale > 10) who demonstrated a total arousal index >or= 10/h and a respiratory disturbance index of < 5/h on prior polysomnography. UARS was definitely diagnosed in patients who demonstrated repetitive increased upper airway resistance (IUAR) associated with brief EEG arousals followed by normalization of esophageal pressure (Pes). IUAR was defined by a pattern of crescendo negative inspiratory Pes of or= 30). Urinary 8-OHdG excretion was significantly higher in the severe OSA group (p = 0.03). Furthermore, urinary 8-OHdG excretion was significantly correlated with parameters of sleep-disordered breathing, including AHI, the apnea index, the oxygen desaturation index (ODI), the duration of oxygen saturation < 90%, and the respiratory arousal index. However, only ODI was significantly correlated with urinary 8-OHdG excretion after adjustment for confounding factors that are considered to be related to oxidative stress. CONCLUSIONS: The severity of OSA is independently associated with oxidative stress. Among various sleep-disordered breathing parameters, ODI is most closely related to oxidative stress. PMID- 15888846 TI - Prevalence and reversibility of pulmonary dysfunction in refractory systemic lupus: improvement correlates with disease remission following hematopoietic stem cell transplantation. AB - AIM: To report the prevalence and reversibility of pulmonary function test (PFT) abnormalities among systemic lupus erythematosus (SLE) patients, refractory to therapy, undergoing hematopoietic stem cell transplantation (HSCT). METHODS: Thirty-four SLE patients received 200 mg/kg cyclophosphamide and 90 mg/kg equine antithymocyte globulin followed by HSCT. PFTs were performed prior to, at 6 months, and yearly following HSCT. RESULTS: The prevalence of significant PFT abnormalities was high (97%). Low FEV(1) and FVC occurred in 26 of 34 patients (76%). A significant abnormality in diffusion capacity of the lung for carbon monoxide (Dlco) occurred in 26 of 32 individuals able to complete Dlco testing (81%). Dlco 18 months after HSCT. Five of 28 patients had a normal entry FVC; for each, the FVC remains normal. Of the 23 patients with an abnormal baseline FVC, 18 have improved, 15 completely and 3 partially. Eight of these 18 patients also have improved Dlco. The two patients with a diagnosis of SLS and one patient with SLE-related pulmonary hypertension improved in both parameters. Only 5 of 23 patients with an abnormal FVC did not improve. Each of these five patients retained active lupus in spite of HSCT. CONCLUSION: The prevalence of lung impairment among SLE patients requiring long-term immune suppression is high. Following HSCT, pulmonary impairments can improve, which is sustained if disease control is sustained. PMID- 15888847 TI - "Ultrasound comet-tail images": a marker of pulmonary edema: a comparative study with wedge pressure and extravascular lung water. AB - BACKGROUND: Echographic examination of the lung surface may reveal multiple "comet-tail images" originating from water-thickened interlobular septa. These images could be useful for noninvasive assessment of interstitial pulmonary edema. STUDY OBJECTIVE: The purpose of this study was to assess the diagnostic accuracy of lung comet-tail images compared with chest radiography, wedge pressure, and extravascular lung water (EVLW) quantified by the indicator dilution method (PiCCO System, version 4.1; Pulsion Medical Systems; Munich, Germany). METHODS AND PATIENTS: We enrolled 20 patients (mean age, 62.6 +/- 11.5 years [+/- SD]). Patients were studied before, immediately after, and 24 h following cardiac surgery with chest ultrasound, chest radiography, pulmonary artery catheterization, and the PiCCO system. Performing echo scanning (right and left hemithorax, from second to fourth intercostal space, from parasternal to midaxillary line), an individual patient comet score was obtained by summing the number of comets in each scanned space. RESULTS: A total of 60 comparisons were obtained. Significant positive linear correlations were found between comet score and EVLW determined by the PiCCO System (r = 0.42, p = 0.001), between comet score and wedge pressure (r = 0.48, p = 0.01), and between comet score and radiologic lung water score (r = 0.60, p = 0.0001). CONCLUSIONS: The presence and the number of comet-tail images provide reliable information on interstitial pulmonary edema. Therefore, ultrasonography represent an attractive, easy-to-use, bedside diagnostic tool for assessing cardiac function and pulmonary congestion. PMID- 15888848 TI - Alveolar nitric oxide and effect of deep inspiration during methacholine challenge. AB - STUDY OBJECTIVES: To assess whether the dual anatomic origin of exhaled nitric oxide (NO), namely alveolar and bronchial, could explain the link between exhaled NO and airway responsiveness, and could participate in the bronchodilatory effect of deep inspiration (DI) that may be evidenced during methacholine challenge. DESIGN AND SETTING: Prospective study in a laboratory performing pulmonary function tests of an academic hospital. PATIENTS AND INTERVENTIONS: Patients underwent multiple flow analysis of exhaled NO, allowing calculation of total maximum airway NO flux (J'awno) and NO concentration of expansible compartment (CAno), and received a cumulative methacholine dose of 2,000 microg. DI effect was assessed by continuous measurement of the resistance of respiratory system using the forced oscillation technique before and after DI. RESULTS: In a first phase involving 23 patients, a positive correlation between log values of J'awno and CAno was demonstrated with the degree of airway responsiveness (percentage of FEV(1) decrease). In a second phase involving 38 patients, only log CAno was correlated with responsiveness, and no significant relationship was demonstrated between J'awno or CAno and the effect of DI. Patients with smaller airways and/or distal airflow limitation exhibited a constrictive response to DI. CONCLUSION: Airway responsiveness is mainly associated with an increase in distal origin of NO output, and no relationship between exhaled NO and the effect of DI was evidenced. PMID- 15888849 TI - Fade of pulmonary function during residual neuromuscular blockade. AB - OBJECTIVES: A decrement in evoked muscle force with repetitive nerve stimulation (fade) suggests impaired neuromuscular transmission. We tested the hypothesis that fade of pulmonary function, ie, a decrease in values of FVC with the second spirometric maneuver compared to the first maneuver, occurs during impaired neuromuscular transmission. DESIGN: Prospective study. PARTICIPANTS: Six healthy male volunteers. INTERVENTIONS: A series of three consecutive spirometric maneuvers was performed every 5 min in six awake healthy volunteers before, during, and after partial paralysis evoked by rocuronium (0.01 mg/kg IV plus 2 to 8 microg/kg/min). MEASUREMENTS AND RESULTS: We measured FVC, FEV(1), forced inspiratory volume in 1 s (FIV(1)), peak expiratory flow (PEF), and peak inspiratory flow (PIF) by spirometry, and force of adductor pollicis muscle by mechanomyography (train-of-four [TOF] stimulation). A statistically significant fade (reduction of the second maneuver from the first maneuver) of FVC, FEV(1), FIV(1), PEF, and PIF was observed during neuromuscular blockade. With peak relaxation (TOF ratio, 0.5) fade amounted to medians of 10% (interquartile range [IQR], 9 to 23%), 7% (IQR, 2 to 16%), 31 (IQR, 19 to 47%), 9% (IQR, 3 to 24%), and 30% (IQR, 5 to 43%), respectively. A fade of >or= 10% was always associated with a clinically relevant (>or= 10%) FVC reduction from baseline (ie, FVC before rocuronium administration). However, FVC reduction from baseline was still present in 23% of measurements without a relevant FVC fade. CONCLUSIONS: A clinically relevant fall (fade) in FVC from the first to the second value during or after neuromuscular blockade suggests impaired pulmonary function and may be due to muscle paralysis. For this reason, the first (best) FVC value may overestimate pulmonary function and expose the patient to an unidentified risk. PMID- 15888850 TI - Chronic idiopathic cough: a discrete clinical entity? AB - STUDY OBJECTIVES: Despite the success of specialist cough clinics, there is increasing recognition of a subgroup of chronic coughers in whom a diagnosis cannot be made even after thorough, systematic investigation. We call this condition chronic idiopathic cough (CIC). The aim of this study is to compare the clinical characteristics of CIC patients with those of coughers in whom a diagnosis has been established (non-CIC) to see if there is a recognizable clinical pattern that distinguishes CIC from non-CIC. DESIGN: Retrospective analysis of the medical records of chronic cough patients. SETTING: The Royal Brompton Hospital Chronic Cough Clinic, London. PATIENTS: One hundred patients with chronic cough referred to the Royal Brompton Hospital Cough Clinic between October 2000 and February 2004. RESULTS: Seventy-one percent of all patients were female. Median age was 57 years (range, 19 to 81 years), with a median duration of symptoms of 48 months (range, 2 to 384 months). The primary diagnoses were CIC (42%), postnasal drip syndromes (22%), gastroesophageal reflux disease (16%), asthma (7%), and others (13%). In CIC patients, the median age at referral, age at onset of cough, and proportion of females did not differ significantly from non-CIC patients. CIC patients had a longer median duration of cough (72 months vs 24 months, p = 0.002), were more likely to report an upper respiratory tract infection (URTI) as the initial trigger of their cough (48% vs 24%, p = 0.0014), and had a significantly lower cough threshold in response to capsaicin (log concentration of capsaicin required to induce five or more coughs, - 0.009 vs 0.592, p = 0.032) than non-CIC patients. CONCLUSIONS: Patients with CIC commonly describe a URTI that initiates their cough, which then lasts for many years, and they demonstrate an exquisitely sensitive cough reflex. We believe that CIC may be a distinct clinical entity with an as-yet unidentified underlying pathology. PMID- 15888851 TI - Noneosinophilic CD4 lymphocytic airway inflammation in menopausal women with chronic dry cough. AB - BACKGROUND: Chronic dry cough without dyspnea and wheezing is a well-known condition that is considered to be clinically overrepresented in women. The etiology and morphology remain unknown in many cases despite thorough investigations. DESIGN: To examine inflammatory cells and the lymphocyte profile in the lower airways and blood in women with chronic cough of unknown etiology. SETTING: University hospital department of respiratory medicine. PARTICIPANTS: Twenty-five otherwise healthy women with idiopathic cough and 11 age-matched healthy control women, all nonatopic nonsmokers. MEASUREMENTS: In order to characterize the cough, a careful standardized interview of the patients was made. Lung functions were tested. Cells were collected by BAL and analyzed for differential cell counts separate in the bronchial (first) wash and in the pooled peripheral washes (BAL fluid). The lymphocyte profile in BAL fluid and blood was characterized by dual-color flow cytometry. RESULTS: Eleven female patients formed a specific group with a history of a dry, nonproductive cough that always started in connection with an airway infection coinciding with menopause. Neither exercise, climate, nor seasonal change influenced the cough. BAL fluid contained an increased number of T (CD3+) lymphocytes: median. Seventy-three percent of T lymphocytes were T-helper lymphocytes (CD4+). A median of 57% of the BAL fluid T cells expressed HLA-DR activation marker compared with a median of 20% in the control subjects and in the other 14 included patients with chronic cough but with minor expectoration periodically (p < 0.001 and p < 0.0001, respectively). No differences between the groups were found in the blood. CONCLUSIONS: HLA-DR activated CD4+ lymphocytic airway inflammation with a low number of eosinophils was identified in a group of nonsmoking, nonatopic otherwise healthy women patients with dry cough of life-long character. The disease appeared exclusively in connection to menopause. PMID- 15888852 TI - Transfusion practice and blood stream infections in critically ill patients. AB - STUDY OBJECTIVE: To examine the relationship between packed RBC (pRBC) transfusion and the development of ICU-acquired bloodstream infections (BSIs) DESIGN: Secondary analysis of a large, prospective, observational study of transfusion practice in critically ill patients. SETTING: A total of 284 adult ICUs in the United States. PATIENTS: Critically ill adults who lacked BSIs both at ICU admission and 48 h after ICU admission. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: BSIs were prospectively tracked in this study, and diagnosis of a new BSI represented the primary end point. Transfusions administered in the ICU prior to development of a BSI were also prospectively recorded. Of 4,892 patients enrolled in this investigation, 3,502 patients lacked BSIs both at ICU admission and 48 h later. Among these individuals, 117 patients (3.3%) had an ICU-acquired BSI. In multivariate analysis adjusting for severity of illness, primary diagnosis, use of mechanical ventilation, placement of central venous catheters, and ICU length of stay, three variables were independently associated with diagnosis of a new BSI: baseline treatment with cephalosporins (odds ratio [OR], 1.84; 95% confidence interval [CI], 1.26 to 2.68), higher sequential organ failure assessment score measured on ICU days 3 to 4 (OR, 1.11; 95% CI, 1.06 to 1.16), and pRBC transfusion (OR, 2.23; 95% CI, 1.43 to 3.52). The relationship between pRBC transfusion and BSI was evident with both small transfusion volumes (OR with 1- to 2-U pRBC transfusion, 1.89; 95% CI, 1.10 to 3.23) and larger transfusion volumes (OR with > 4-U pRBC transfusion, 2.63; 95% CI, 1.52 to 4.53). CONCLUSIONS: pRBC transfusion is associated with subsequent ICU-acquired BSI. Avoiding unnecessary transfusions may decrease the incidence of BSIs. PMID- 15888853 TI - A multidisciplinary community hospital program for early and rapid resuscitation of shock in nontrauma patients. AB - OBJECTIVE: To determine the effect of a community hospital-wide program enabling nurses and prehospital personnel to mobilize institutional resources for the treatment of patients with nontraumatic shock. DESIGN: Historically controlled single-center study. SETTING: A 180-bed community hospital. PATIENTS: Patients in shock who were candidates for aggressive therapy. INTERVENTIONS: From January 1998 to May 31, 2000, patients in shock received standard therapy (control group). During the month of June 2000, intensive education of all health-care providers (ie, prehospital personnel, nurses, and physicians) took place. From July 1, 2000, through June 30, 2001, patients in shock (protocol group) were managed with a hospital-wide shock program. The program included early recognition of shock and the initiation of therapy by nonphysicians. Frontline personnel mobilized a shock team, which used goal-directed resuscitation protocols, early intensivist involvement, and rapid transfer to the ICU where protocols specific to shock etiology were implemented. MEASUREMENTS AND MAIN RESULTS: Eighty-six and 103 patients, respectively, were enrolled in the control and protocol groups. Baseline characteristics were similar. The protocol group had significant reductions in the median times to interventions, as follows: intensivist arrival, 2:00 h to 50 min (p < 0.002); ICU/operating room admission, 2 h 47 min to 1 h 30 min (p < 0.002); 2 L fluid infused, 3 h 52 min to 1 h 45 min (p < 0.0001); and pulmonary artery catheter placement, 3 h 50 min to 2 h 10 min (p 0.02). Good outcomes (ie, discharged to home or to a rehabilitation center) were more likely in the protocol group than in the control group (p = 0.02). The hospital mortality rate was 40.7% in the control group and 28.2% in the protocol group (p = 0.035). CONCLUSION: Similar to current practice in patients who have experienced trauma or cardiac arrest, the empowerment of nonphysician providers to mobilize hospital resources for the care of patients with shock is effective. A community hospital program incorporating the education of providers, the activation of a coordinated team response, and early goal-directed therapy expedited appropriate treatment and was temporally associated with improved outcomes. Randomized multicenter trials are needed to further assess the impact of the shock program on outcomes. PMID- 15888854 TI - Limitations of transcutaneous carbon dioxide measurements for assessing long-term mechanical ventilation. AB - STUDY OBJECTIVES: Transcutaneous CO(2) pressure (Ptcco(2)) and transcutaneous O(2) pressure (Ptco(2)) measurements are routinely used in pediatric ICUs in order to avoid serial arterial punctures. The aim of this study was to determine the value of Ptcco(2) assessment during the evaluation of home ventilation in 12 adult patients with COPD or restrictive respiratory failure in the stable state (mean [+/- SD] basal Paco(2), 48.8 +/- 8.3 mm Hg) who were treated by mask or tracheotomy-mediated ventilation. METHODS: After radial catheter insertion, patients were instructed to breathe spontaneously for 40 min and then to receive ventilation for 40 min according to their individual home ventilation modalities. An in vivo calibration was performed in the initial stage of the study in order to optimize the arterial Pco(2) and Ptcco(2) values. Every 5 min, transcutaneous measurements were performed and simultaneously compared with arterial values. MEASUREMENTS AND RESULTS: Ptcco(2) and Ptco(2) were correlated with arterial values (p < 0.0001) except for Paco(2) values of > 56 mm Hg and Pao(2) values of > 115 mm Hg. During ventilation, Paco(2) decreased >or= 4 mm Hg in seven patients. Ptcco(2) variations recorded during consecutive 5-min periods while the patient received mechanical ventilation were well correlated with the arterial variations (p = 0.0033), with a delay of < 5 min. CONCLUSION: Ptcco(2) values and variations accurately reflected Paco(2) values and variations during mechanical ventilation. However, the accuracy of these data seems to be restricted to patients with Paco(2) values of < 56 mm Hg. PMID- 15888855 TI - Agreement between bedside blood and plasma glucose measurement in the ICU setting. AB - STUDY OBJECTIVE: To assess the agreement between bedside glucose (bGlu) measurements and laboratory plasma glucose (pGlu) measurements in the ICU setting. DESIGN: Retrospective study. SETTING: ICU of a tertiary academic medical center. PATIENTS: One hundred ninety-seven patients admitted to the ICU from January through December 2002 who underwent 816 simultaneous bGlu and pGlu measurements. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: To calculate the agreement between the two methods of glucose measurement, the mean difference was obtained, and the limits of the agreement were calculated as the mean difference +/- 2 SDs. On 767 occasions, the mean bGlu was 159 mg/dL and the mean pGlu was 151 mg/dL (p < 0.001). The mean difference between the two techniques was 7.9 mg/dL (SD, 17.6 mg/dL), and the limits of agreement were + 43.1 and -27.2. On 31 occasions, the bGlu was reported as > 400 mg/dL, and in these cases the mean pGlu was 423 mg/dL (range, 300 to 900 mg/dL). On 18 occasions, the bGlu was reported as < 50 mg/dL, and in these cases the mean pGlu was 66.9 mg/dL (range, 13 to 198 mg/dL). CONCLUSIONS: On average, bGlu provides a reasonable estimate for pGlu. However, for the individual patient, bGlu gives an unreliable estimate for pGlu. All of those taking care of critically ill patients should be aware of the limitations of bedside glucometry. PMID- 15888856 TI - Management of community-acquired pneumonia in the home: an American College of Chest Physicians clinical position statement. AB - The number of patients with community-acquired pneumonia (CAP) who are being treated at home is increasing for a variety of reasons. These reasons include the increased availability and cost considerations of oral antibiotics that have been shown to be effective, as well as the consideration of patient and family preferences. However, there is still considerable variability in strategies for the management of patients with CAP. This American College of Chest Physicians position statement, which was cosponsored by the American Academy of Home Care Physicians, provides recommendations on the various aspects of home care for patients with this condition. Included are recommendations for evaluation and diagnosis in the home environment and the determination of the site of care, and an outline of an in-home management plan. The position statement also provides recommendations for issues related to patient and caregiver commitment to the plan, and for monitoring and follow-up. Recommendations are directed toward immunocompetent adult patients with CAP who are at home or in other unskilled residential facilities. These patients can include previously healthy individuals or chronically ill individuals who choose not to go to the hospital, or hospitalized patients who are completing a hospital discharge plan. The recommendations in this statement take into consideration the best course of action for the patient, as determined by incorporating the most recent evidence with clinician judgment and patient preferences. These recommendations also consider the available resources. Therefore, these recommendations may not apply to every patient, and interventions may need to be structured based on the individual. In addition to providing recommendations for the home care management of patients with CAP, we hope that this clinical policy statement will alert readers to the need for more scientific evidence related to the clinical and psychosocial issues associated with managing this condition. PMID- 15888857 TI - Outcomes of elderly survivors of intensive care: a review of the literature. AB - An increasing proportion of critically ill patients are elderly (ie, >or= 65 years of age). This poses complex challenges and choices for the management of elderly patients. Outcome following admission to the ICU has been traditionally concerned with mortality. Beyond mortality, outcomes such as functional status and health-related quality of life (HRQOL) have assumed greater importance. This article reviews the literature, published in English from 1990 to December 2003, pertaining to HRQOL and functional status outcomes of elderly patients. Functional status and HRQOL of elderly survivors of ICUs has been underinvestigated. There is no agreement as to the optimal instrument choice, and differences between studies preclude meaningful comparison or pooling of results. PMID- 15888858 TI - Quality of dying and death in two medical ICUs: perceptions of family and clinicians. AB - OBJECTIVE: We compared perceptions of the quality of dying and death in the ICU across nurses, resident physicians, attending physicians, and family members. The aim was to obtain a surrogate assessment of the quality of the dying process and examine differences in the perceptions of different types of raters. DESIGN: Cross-sectional survey of family members and ICU clinicians conducted following the death of enrolled patients. SETTING: Two medical ICUs at academic tertiary care medical centers. PATIENTS: Patients dying in the ICU (n = 68). MEASUREMENTS AND RESULTS: The previously validated Quality of Dying and Death (QODD) instrument was modified for use in the ICU. Within 48 h of the time of death, the nurse, resident, and attending physician caring for the patient were asked to complete the QODD. One month following the death, a designated family member was contacted and the QODD was administered on the telephone. Family members and attending physicians gave the most favorable ratings of death, while nurses and residents provided less favorable ratings. Significant differences between these groups were notable (p < 0.01) on items related to patient autonomy: maintaining dignity, being touched by loved ones, and the overall quality of death. CONCLUSIONS: The perception of dying and death in the ICU varies considerably between nurses, attending physicians, resident physicians, and family members. Further studies are needed to explain these differences and determine the utility of the ICU QODD instrument for assessing and improving the quality of end-of-life care in the ICU. PMID- 15888859 TI - Performance characteristics of 10 home mechanical ventilators in pressure-support mode: a comparative bench study. AB - OBJECTIVE: Inspiratory pressure (Pi) support delivered by a bilevel device has become the technique of choice for noninvasive home ventilation. Considerable progress has been made in the performance and functionality of these devices. The present bench study was designed to compare the various characteristics of 10 recently developed bilevel Pi devices under different conditions of respiratory mechanics. DESIGN: Bench model study. SETTING: Research laboratory, university hospital. MEASUREMENTS: Ventilators were connected to a lung model, the mechanics of which were set to normal, restrictive, and obstructive, that was driven by an ICU ventilator to mimic patient effort. Pressure support levels of 10 and 15 cm H(2)O, and maximum were tested, with "patient" inspiratory efforts of 5, 10, 15, 20, and 25 cm H(2)O. Tests were conducted in the absence and presence of leaks in the system. Trigger delay, trigger-associated inspiratory workload, pressurization capabilities, and cycling were analyzed. RESULTS: All devices had very short trigger delays and triggering workload. Pressurization capability varied widely among the machines, with some bilevel devices lagging behind when faced with a high inspiratory demand. Cycling was usually not synchronous with patient inspiratory time when the default settings were used, but was considerably improved by modifying cycling settings, when that option was available. CONCLUSIONS: A better knowledge of the technical performance of bilevel devices (ie, pressurization capabilities and cycling profile) may prove to be useful in choosing the machine that is best suited for a patient's respiratory mechanics and inspiratory demand. Clinical algorithms to help set cycling criteria for improving patient-ventilator synchrony and patient comfort should now be developed. PMID- 15888860 TI - Bone marrow-derived cells contribute to pulmonary vascular remodeling in hypoxia induced pulmonary hypertension. AB - STUDY OBJECTIVE: In these days, it was reported that bone marrow (BM) cells might take part in the remodeling of some systemic vascular diseases; however, it remains unknown whether the BM cells were involved in the vascular remodeling of pulmonary arteries and the progression of pulmonary hypertension (PH). The purpose of this study was to investigate whether BM-derived cells contribute to pulmonary vascular remodeling in hypoxia-induced PH. MATERIALS AND METHODS: To investigate the role of BM-derived cells, we transplanted the whole BM of enhanced green fluorescent protein (GFP)-transgenic mice to the lethally irradiated syngeneic mice (n = 30). After 8 weeks, chimera mice were exposed to consistent hypoxia using a hypoxic chamber (10% O(2)) for up to 4 or 8 weeks (10 mice per group). After hemodynamics and the ratio of right ventricular (RV) weight to left ventricle (LV) weight, RV/(LV + septum [S]), were measured, histologic and immunofluorescent staining were performed. RESULTS: BM transplanted mice showed a high chimerism (mean [+/- SEM], 91 +/- 2.3%). RV systolic pressure and the RV/(LV + S) ratio increased significantly with time in PH mice, indicating RV hypertrophy. Marked vascular remodeling including medial hypertrophy and adventitial proliferation was observed in the pulmonary arteries of PH mice. Strikingly, a number of GFP(+) cells were observed at the pulmonary arterial wall, including the adventitia, in hypoxia-induced PH mice, while very few cells were observed in the control mice. Metaspectrometer measurements using confocal laser scanning microscopy confirmed that this green fluorescence was produced by GFP, suggesting that these GFP(+) cells were mobilized from the BM. Most of them expressed alpha-smooth muscle actin, a smooth muscle cell, or myofibroblast phenotype, and contributed to the pulmonary vascular remodeling. A semiquantitative polymerase chain reaction of the GFP gene revealed that the BM derived GFP-positive cells in the PH group were observed more than eightfold as often compared with the control mice. CONCLUSION: The BM-derived cells mobilize to the hypertensive pulmonary arteries and contribute to the pulmonary vascular remodeling in hypoxia-induced PH mice. PMID- 15888862 TI - Direct visual guidance for chest tube placement through a single-port thoracoscopy: a novel technique. PMID- 15888861 TI - Sentinel lymph node mapping of the pleural space. AB - STUDY OBJECTIVES: Although the sentinel lymph node (SLN) concept has traditionally been applied to solid organs, we hypothesized that the pleural space might drain into a specific SLN group. The identification of such a nodal group could assist in the staging and treatment of pleural-based diseases, such as mesothelioma, or other lung cancers with visceral pleural invasion. The purpose of this study was to determine whether the pleural space has an SLN group. DESIGN: Sixteen rats underwent right or left pleural space injection of a novel lymph tracer, quantum dots (QDs), which have a hydrodynamic diameter of 15 nm and fluoresce in the near-infrared (NIR) spectrum. Nodal uptake of the entire thorax was imaged with a custom system that simultaneously acquired color video, NIR fluorescence of the QDs, and a merged picture of the two in real-time. Six pigs underwent right or left pleural space injection of QDs and similar imaging. MEASUREMENTS AND RESULTS: In the rat, the QDs drained solely to the highest superior mediastinal lymph node group, corresponding to lymph node station 1, according the regional lymph node classification for lung of the American Joint Committee on Cancer. In one rat, the injection of QDs in the left pleural space resulted in migration to the contralateral station 1 lymph node group. The injection of QDs in the right or left pleural space of the pig resulted in migration solely to the ipsilateral highest superior mediastinal lymph node group. CONCLUSIONS: NIR fluorescence imaging in two species demonstrated that the highest superior mediastinal lymph nodes of station 1 are the SLNs of the pleural space. This study also provides intraoperative feasibility and proof of the concept for identifying lymph nodes communicating with the pleural space on a patient-specific basis, in real-time, and with high sensitivity. PMID- 15888863 TI - Hypocapnia and asthma: a mechanism for breathing retraining? AB - There is some evidence that breathing retraining may be beneficial for patients with asthma, but the mechanism behind this benefit is still unknown. One hypothesis is that individuals can be trained to raise carbon dioxide levels and thereby reverse the bronchoconstrictive effects of hypocapnia and utilize the bronchodilatory effects of hypercapnia. This theory presupposes that individuals with asthma have lower carbon dioxide levels than the healthy population. This article reviews the available evidence supporting the hypothesis and concludes that although attractive, there is currently insufficient evidence to attribute the benefits of breathing retraining to this mechanism. PMID- 15888864 TI - Hyperventilation in head injury: a review. AB - The aim of this review was to consider the effects of induced hypocapnia both on systemic physiology and on the physiology of the intracranial system. Hyperventilation lowers intracranial pressure (ICP) by the induction of cerebral vasoconstriction with a subsequent decrease in cerebral blood volume. The downside of hyperventilation, however, is that cerebral vasoconstriction may decrease cerebral blood flow to ischemic levels. Considering the risk-benefit relation, it would appear to be clear that hyperventilation should only be considered in patients with raised ICP, in a tailored way and under specific monitoring. Controversy exists, for instance, on specific indications, timing, depth of hypocapnia, and duration. This review has specific reference to traumatic brain injury, and is based on an extensive evaluation of the literature and on expert opinion. PMID- 15888865 TI - Use of recombinant activated factor VII for bleeding following operations requiring cardiopulmonary bypass. AB - Postoperative bleeding is a common complication following cardiothoracic surgical procedures requiring cardiopulmonary bypass (CPB). Serious bleeding complications requiring the administration of blood products, hemostatic drugs, and even repeat surgery are associated with considerable morbidity, mortality, and resource consumption. Therapy with recombinant activated factor VII (rFVIIa) may be an effective treatment strategy for patients with refractory bleeding. We report the successful use of rFVIIa for the treatment of intractable postoperative bleeding following aortic aneurysm repair in two patients with Marfan syndrome. In both patients, surgical reexploration was avoided, and the patients' clinical status was stabilized after the administration of rFVIIa. In one patient, hemostasis was rapidly achieved within minutes, whereas hemostasis occurred gradually over several hours in the second patient. Including our personal experience with the two cases, the use of rFVIIa has been reported in 20 patients who required CPB for cardiothoracic surgical procedures. Hemostasis was achieved in all patients. In 14 patients (70%), rapid hemostasis was achieved following a single dose of rFVIIa (mean dose, 57 microg/kg). In the remaining six patients, gradual hemostasis was achieved after a mean of 3.4 doses (mean cumulative dose, 225 microg/kg). Two patients (10%) were believed to have experienced thromboembolic complications after the administration of rFVIIa (one was fatal), and, in another patient, intracoronary thrombosis was suspected but was not confirmed. In patients experiencing postoperative bleeding complications that are refractory to treatment with blood products, hemostatic agents, and/or repeat surgery, the use of rFVIIa may be considered. PMID- 15888866 TI - Pulmonary arterial hypertension: the key role of echocardiography. AB - Given the nonspecific nature of its early symptoms and signs, pulmonary arterial hypertension (PAH) is often diagnosed in its advanced stages. Although clinical assessment is essential when initially evaluating patients with suspected PAH, echocardiography is a key screening tool in the diagnostic algorithm. It not only provides an estimate of pulmonary pressure at rest and during exercise, but it may also help to exclude any secondary causes of pulmonary hypertension, predict the prognosis, monitor the efficacy of specific therapeutic interventions, and detect the preclinical stage of the disease. PMID- 15888867 TI - The insidious onset of dyspnea and right lung collapse in a 35-year-old man. PMID- 15888868 TI - Growing pulmonary nodule with increased 18-fluorodeoxyglucose uptake in a former smoker. PMID- 15888869 TI - Familial solitary fibrous tumor of the pleura: a case report. AB - This report describes the occurrence of solitary fibrous tumors of the pleura in a mother and her daughter. No other occurrence of this rare tumor in members of the same family has ever been reported. PMID- 15888870 TI - Silicone thorax due to a ruptured breast implant. AB - A woman with a history of bilateral mastectomy and silicone implants for fibrocystic disease and a history of atrial septal defect repair presented with pleural nodules on a chest radiograph. A thorascopic biopsy performed for possible mesothelioma demonstrated chronic inflammation and focal pleural fibrosis due to a foreign-body reaction secondary to silicone. This was confirmed using scanning electron microscopy and energy-dispersive radiograph elemental analysis. As the population ages, the increasing frequency of ruptured silicone implants and the need for heart surgery may result in a corresponding increase in the risk for fibrothorax secondary to inadvertent silicone introduction during surgery. PMID- 15888871 TI - Heparin-induced skin lesions and other unusual sequelae of the heparin-induced thrombocytopenia syndrome: a nested cohort study. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is caused by platelet activating, heparin-dependent IgG antibodies (HIT-IgG). Although HIT is known to predispose the patient to thrombosis, the relationship between the formation of HIT-IgG and various other unusual clinical sequelae putatively linked with the HIT syndrome, such as heparin-induced skin lesions and acute anaphylactoid reactions following treatment with an IV heparin bolus, is not clear. METHODS: We used data from a clinical trial of postoperative heparin prophylaxis to compare the frequency of one or more predefined unusual clinical sequelae developing in 20 patients who formed platelet-activating HIT-IgG with 80 control patients who did not form HIT-IgG (nested cohort study). RESULTS: Five of the 20 patients in whom HIT-IgG developed had one or more unusual clinical sequelae, compared with none of 80 control patients (25% vs 0%, respectively; odds ratio, infinity; 95% confidence interval, 4.3 to infinity; p < 0.001). The unusual complications included heparin-induced erythematous or necrotic skin lesions (n = 4), an anaphylactoid reaction following IV heparin bolus use (n = 1), and warfarin associated venous limb ischemia (n = 1). Thrombocytopenia, as it is conventionally defined (ie, platelet count fall to < 150 x 10(9) cells/L) developed in only one of these five patients. CONCLUSIONS: Certain unusual clinical sequelae, such as heparin-induced skin lesions, are strongly associated with the formation of HIT-IgG and should be considered as manifestations of the HIT syndrome, even in the absence of thrombocytopenia as conventionally defined. PMID- 15888872 TI - Poor quality of life in young smokers with smoking history of short duration. PMID- 15888873 TI - Missed opportunity to address the critical care medicine crisis. PMID- 15888874 TI - Yoga and pneumothorax. PMID- 15888875 TI - Treatment outcome in Mycobacterium avium pulmonary disease: a correction and comment. PMID- 15888876 TI - Chylothorax in hematologic malignancies. PMID- 15888877 TI - Negative pressure pulmonary edema as an unsuspected imitator of acute lung injury/ARDS. PMID- 15888878 TI - CO2 retention in acute severe asthma. PMID- 15888879 TI - Tumor necrosis factor-alpha in parapneumonic effusion. PMID- 15888880 TI - Auscultation and Don Quixote. PMID- 15888881 TI - The importance of clinical registries for pulmonary epithelioid hemangioendothelioma. PMID- 15888882 TI - Epoprostenol and nesiritide in pulmonary hypertension. PMID- 15888883 TI - Dairy farm effluent effects on urine patch nitrous oxide and carbon dioxide emissions. AB - Dairy farm effluent (DFE) comprises animal feces, urine, and wash-down water collected at the milking shed. This is collected daily during the milking season and sprayed onto grazed dairy pastures. Urine patches in grazed pastures make a significant contribution to anthropogenic N(2)O emissions. The DFE could potentially mitigate N(2)O emissions by influencing the N(2)O to dinitrogen (N(2)) ratio, since it contains water-soluble carbon (WSC). Alternatively, DFE may enhance N(2)O emissions from urine patches. The application of DFE may also provide a substrate for the production of CO(2) in pasture soils. The effects of DFE on the CO(2) and N(2)O emissions from urine patches are unknown. Thus a laboratory experiment was performed where repeated DFE applications were made to repacked soil cores. Dairy farm effluent was applied at 0, 7, or 14 d after urine deposition. The urine was applied once on Day 0. Urine contained (15)N-enriched urea. Measurements of N(2)O, N(2), and carbon dioxide (CO(2)) fluxes, soil pH, and soil inorganic N concentrations were made. After 43 d the DFE had not mitigated N(2)O fluxes from urine patches. A small increase in the N(2)O flux occurred from the urine-treated soils where DFE was applied 1 wk after urine deposition. The amount of WSC applied in the DFE proved to be insignificant compared with the amount of soil C released as CO(2) following urine application. The priming of soil C in urine patches has implications for the understanding of soil C processes in grazed pasture ecosystems and the budgeting of C within these ecosystems. PMID- 15888884 TI - Rapid manganese removal from mine waters using an aerated packed-bed bioreactor. AB - In the UK, the Environmental Quality Standard for manganese has recently been lowered to 30 microg/L (annual average), which is less than the UK Drinking Water Inspectorate's Maximum Permitted Concentration Value (50 microg/L). Current passive treatment systems for manganese removal operate as open-air gravel-bed filters, designed to maximize either influent light and/or dissolved oxygen. This requires large areas of land. A novel enhanced bioremediation treatment system for manganese removal has been developed that consists of a passively aerated subsurface gravel bed. The provision of air at depth and the use of catalytic substrates help overcome the slow kinetics usually associated with manganese oxidation. With a residence time of only 8 h and an influent manganese concentration of approximately 20 mg/L, >95% of the manganese was removed. The treatment system also operates successfully at temperatures as low as 4 degrees C and in total darkness. These observations have positive implications for manganese treatment using this technique in both colder climates and where large areas of land are unavailable. Furthermore, as the operation of this passive treatment system continually generates fresh manganese oxyhydroxide, which is a powerful sorbent for most pollutant metals, it potentially has major ancillary benefits as a removal process for other metals, such as zinc. PMID- 15888885 TI - Initial tree establishment on blocky quarry waste ameliorated with hydrogel or slate processing fines. AB - Pocket planting reclamation techniques developed in the 1970s for revegetating blocky quarrying waste have met with very limited success, often because the low water-holding capacity of the waste and limited root development within a small volume of planting pocket material result in severe drought mortality. We tested pocket planting approaches for waste tip reclamation at Europe's largest slate quarry, and compared materials for enhancing the continuity of water- and nutrient-holding down into the interior of the waste tip. When small compost filled pocket planting bags were placed above slate processing fines (SPF) or water absorbent cross-linked polyacrylamide gel ("hydrogel"), tree growth rates increased in comparison with pocket planting bags alone. The SPF significantly improved tree survival especially during severe drought, but survival was not enhanced by the use of hydrogel. The sorption characteristics of hydrogel indicated that its presence may help to reduce nutrient leaching, but that it may have a negative effect on nitrogen availability. A more likely explanation for the poor performance of pure hydrogel is that it did not maintain sufficient available water, because of discontinuities caused by shrinkage and movement of the hydrogel, and/or degradation of water-holding capacity with environmental exposure. However, the root growth observed in the hydrogel treatments suggests that this technique, if adapted to reduce the effects of hydrogel shrinkage by using finer-grade hydrogel, mixing it with other soil-forming material, and reducing its exposure to extremes of temperature or sunlight, might have the potential to improve the growth and survival of trees planted on sites where delivery of heavy materials such as SPF is impractical. Fine mineral processing waste is freely available at active quarries and should be seen as a key resource for reclamation schemes. PMID- 15888886 TI - The acetochlor registration partnership: prospective ground water monitoring program. AB - The Acetochlor Registration Partnership conducted a prospective ground water (PGW) monitoring program to investigate acetochlor [2-chloro-N-(ethoxymethyl)-N (2-ethyl-6-methylphenyl)-acetamide] transport to ground water at eight sites. The distribution of soil textures among these sites was weighted toward coarser soil types, while also including finer-textured soils that dominate most corn (Zea mays L.)-growing areas of the United States. Each site consisted of a 1.2-ha test plot adjacent to a 0.2-ha control plot. Suction lysimeters and monitoring wells were installed at multiple depths within each test and control plot to sample soil-pore water and near-surface ground water. Irrigation was applied to each site during the growing season to ensure water input of 110 to 200% of average historical rainfall. Acetochlor dissipated rapidly from surface soils at all sites with a DT(50) (time for 50% of the initial residues to dissipate) of only 3 to 9 d, but leaching was not an important loss mechanism, with only 0.25% of the 15,312 soil-pore water and ground water samples analyzed containing parent acetochlor at or above 0.05 microg L(-1). However, quantifiable residues of a soil degradation product, acetochlor ethanesulfonic acid, were more common, with approximately 16% of water samples containing concentrations at or above 1.0 microg L(-1). A second soil degradation product, acetochlor oxanilic acid, was present at concentrations at or above 1.0 microg L(-1) in only 0.15% of water samples analyzed. The acetochlor PGW program demonstrated that acetochlor lacks the potential to leach to ground water at detectable concentrations, and when applied in accordance with label restrictions, is unlikely to move to ground water at concentrations hazardous to human health. PMID- 15888887 TI - Sources and evolution of anthropogenic lead in dated sediments from Lake Clair, Quebec, Canada. AB - Two sediments cores were collected from the deepest part of Lake Clair (Quebec, Canada) to assess the historical sources of Pb additions to the lake. The cores were collected by divers by carefully inserting a Plexiglas tube into the sediments. To determine the stratigraphic ages of the sediments, (210)Pb and (137)Cs activities were counted by gamma-ray spectroscopy. Lead concentrations and isotopic ratios were performed by inductively coupled plasma-mass spectrometry (ICP-MS), following digestion of the samples with a mixture of HF, HNO(3), and HClO(4) acids and Pb separation by anion-exchange chromatography. Starting at the middle of the 19th century, Pb content of the sediments increased until 1975. The maximum Pb enrichment factor of 35 times (relative to the natural background) was found in sediments deposited in 1975. At this time, excess Pb flux was estimated to be about 0.03 g m(-2) yr(-1). Before 1872, the Pb isotopic ratios were relatively stable (mean (206)Pb/(207)Pb = 1.20 +/- 0.01), reflecting the natural Pb background. Between 1872 and 1894, the source of anthropogenic Pb was highly radiogenic as shown by the Pb isotopic signatures of the sediments (mean (206)Pb/(207)Pb = 1.22 +/- 0.01), possibly reflecting deforestation and agricultural developments in the St.-Lawrence Valley. Between 1894 and 1937, widespread use of industrial and domestic charcoals may explain the isotopic composition of Pb accumulated in the sediments (mean (206)Pb/(207)Pb = 1.19 +/- 0.01). From 1937 to 1975, Pb isotopic compositions became less radiogenic ((206)Pb/(207)Pb from 1.18 to 1.17) even though elemental Pb abundance reached extremely high values (623 mg kg(-1)). This isotopic shift reflects increased use of alkyl-lead in gasoline. For sediments accumulated between 1967 and 1996, the U.S. contribution to anthropogenic Pb accumulated in Lake Clair sediments amounted to between 30 and 63%. PMID- 15888888 TI - A regional-scale study on the crop uptake of cadmium from sandy soils: measurement and modeling. AB - Plant uptake is one of the major pathways by which cadmium (Cd) in soils enters the human food chain. This study was conducted to investigate the uptake of Cd by crops from soils within the wastewater irrigation area (WIA) of Braunschweig (Germany) and to develop a simple process-oriented model that is suited to predict Cd uptake at the regional scale. The sandy soils within the WIA (4300 ha) have received considerable loads of heavy metals by irrigation using municipal wastewater for up to 40 years. In 1998 and 1999, we sampled soil and plant material at 40 potato (Solanum tuberosum L.), 40 sugar beet (Beta vulgaris L.), and 32 winter wheat (Triticum aestivum L.) fields. In both years and for all three crops, we found close linear relationships between the Cd content of plant material and the Cd concentration in soil solution. For all three crops, we observed a trend of relatively increased Cd uptake in the year with the higher saturation deficit of the atmosphere. We interpret this to indicate that transpiration plays an important role in the Cd uptake of crops under the conditions of the WIA. In modeling the uptake of Cd by crops, we assume that uptake is proportional to mass flow, that is, the product of water transpired, Cd concentration in soil solution, and a plant-specific empirical parameter. The simulations agreed well with the observed Cd contents in crops. Our model explained between 66 and 87% of the observed variance. PMID- 15888889 TI - Green roof stormwater retention: effects of roof surface, slope, and media depth. AB - Urban areas generate considerably more stormwater runoff than natural areas of the same size due to a greater percentage of impervious surfaces that impede water infiltration. Roof surfaces account for a large portion of this impervious cover. Establishing vegetation on rooftops, known as green roofs, is one method of recovering lost green space that can aid in mitigating stormwater runoff. Two studies were performed using several roof platforms to quantify the effects of various treatments on stormwater retention. The first study used three different roof surface treatments to quantify differences in stormwater retention of a standard commercial roof with gravel ballast, an extensive green roof system without vegetation, and a typical extensive green roof with vegetation. Overall, mean percent rainfall retention ranged from 48.7% (gravel) to 82.8% (vegetated). The second study tested the influence of roof slope (2 and 6.5%) and green roof media depth (2.5, 4.0, and 6.0 cm) on stormwater retention. For all combined rain events, platforms at 2% slope with a 4-cm media depth had the greatest mean retention, 87%, although the difference from the other treatments was minimal. The combination of reduced slope and deeper media clearly reduced the total quantity of runoff. For both studies, vegetated green roof systems not only reduced the amount of stormwater runoff, they also extended its duration over a period of time beyond the actual rain event. PMID- 15888890 TI - Measuring sorption of hydrophilic organic compounds in soils by an unsaturated transient flow method. AB - Determination of sorption of hydrophilic, weakly sorbing organic compounds in soil by conventional batch methods using a slurried suspension is often prone to considerable errors because small changes in the solution concentration on equilibration must be accurately determined. This difficulty is exacerbated for compounds susceptible to degradation, which also decreases the solution concentration. The objective of this study was to determine sorption of hydrophilic pesticides by applying an unsaturated transient flow method, which enables determination of sorption at sufficiently small solution to soil ratios. The method makes use of piston-like displacement of the antecedent solution in equilibrium with sorbed phase when pesticide-free water is infiltrated into a soil column spiked with a pesticide. Pesticide sorption and the solution concentration are inferred from a plot of total pesticide content per unit mass of soil vs. water content in a region where the antecedent solution is accumulated. Thus, extraction of solution from relative dry soil is unnecessary. We tested this method for two hydrophilic pesticides, monocrotophos [dimethyl (E) 1-methyl-2-(methyl-carbamoyl) vinyl phosphate] and dichlorvos (2,2-dichlorovinyl dimethyl phosphate). The sorption coefficient, K(d), obtained for monocrotophos was slightly lower than that by batch method (K(d) = 0.10 vs. 0.19 L kg(-1)), whereas for dichlorvos, a compound highly susceptible to degradation, the unsaturated flow method yielded a much smaller K(d) (0.19 vs. 3.22 L kg(-1)). The K(d) values for both compounds were consistent with the observed retardation in the pesticide displacement in the columns. The proposed method is more representative of field conditions and particularly suitable for weakly sorbing organic compounds in soils. PMID- 15888891 TI - The influence of lipids on the energetics of uptake of polycyclic aromatic hydrocarbons by natural organic matter. AB - Although most of the organic carbon in soils and sediments may be composed of humic substances, their interaction with other compounds, especially their sorption interactions, may be significantly affected by the presence of small amounts of the other components of natural organic matter (NOM). In this investigation, the influence of the lipid fraction of NOM on the sorption thermodynamics of fluorene, phenanthrene, and pyrene to several geosorbent samples was examined before and after extraction of lipids. Batch experiments were performed at the same concentration for all polycyclic aromatic hydrocarbons (PAHs) (0.025 x their solubility in water) at different temperatures (10, 20, 30, and 40 degrees C), and the thermodynamic parameters were calculated. Removal of the lipids increases the sorption capacity of the samples as well as the exothermicity of the process. The free energy change was negative for all the samples and no significant differences were noticed on lipid removal. The entropy changes were small and positive for the whole geosorbent samples, but even smaller or more negative when the lipids were removed. This indicates that the interaction of PAHs with soils and sediments in the absence of extractable lipids is stronger and the mechanisms involved may be different, changing from a partitioning-like mechanism to specific adsorption. Because of the competition between lipids and PAHs for the same sorption sites, the lipids can be viewed as an "implicit sorbate." PMID- 15888892 TI - A thermodynamically based method to quantify true sorption hysteresis. AB - Sorption of organic chemicals to soils and sediments often shows true hysteresis (i.e., nonsingularity of the sorption-desorption isotherm not attributable to known experimental artifacts). Since true sorption hysteresis is fundamentally important to contaminant fate, a way to quantify it is desirable. Previously proposed indices of hysteresis are empirical and usually depend on the isotherm model. True sorption hysteresis to synthetic and natural organic solids has been attributed to irreversible alteration of the solid during the sorption-desorption cycle. Given this mechanism, we propose the Thermodynamic Index of Irreversibility (TII) for quantifying hysteresis in soils where natural organic matter dominates the sorption process. The TII is based on the difference in free energy between the real desorption state and the hypothetical fully reversible state. The index is 0 for completely reversible systems and approaches 1 as the process tends toward complete irreversibility. It does not require any assumptions about the physical properties or molecular composition of the solid, and it does not depend on a specific equilibrium model. A sensitivity analysis of measurement errors provides general recommendations for the setup of sorption desorption experiments. The TII was applied to sorption of 1,4-dichlorobenzene (DCB) to two high-organic soils, Pahokee peat (PP) and Amherst soil (AS), and a low-rank coal reference material, Beulah-Zap lignite (BZL). Common artificial causes of hysteresis were eliminated. Hysteresis was significant in the peat and the coal. The TII was clearly concentration dependent for both solids; it decreased with concentration for the peat, but increased with concentration for the coal. The TII allows quantification of hysteresis as a function of sorbate sorbent combination, concentration, time, and other variables. PMID- 15888893 TI - Effect of petroleum-containing wastewater irrigation on bacterial diversities and enzymatic activities in a paddy soil irrigation area. AB - Effects of petroleum contamination on bacterial diversities and enzymatic activities in paddy soils were investigated in the Shenfu irrigation area, the largest area irrigated by oil-containing wastewater for more than 50 yr in northeastern China. Bacterial diversities were determined by conventional colony morphology typing techniques and 16S rDNA polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE). Dehydrogenase, hydrogen peroxidase, polyphenol oxidase, urease, and substrate-induced respiration (SIR) were measured to evaluate the effects of petroleum-containing wastewater irrigation on soil biochemical characteristics. Results showed that paddy soil total petroleum hydrocarbon (TPH) concentration in the irrigation area varied from 277.11 to 5213.37 mg kg(-1) dry soil. Soil TPH concentration declined along the gradient of the irrigation channel from up- to downstream. At the current pollution level, the paddy soil TPH concentration was positively correlated with the colony forming units (CFU) of aerobic heterotrophic bacteria (AHB) (r = 0.928, p < 0.001) and the genetic diversity based on DGGE profiles (r = 0.655, p < 0.05). The bacterial diversities in the soils based on colony morphotypes of AHB also increased with TPH concentration (r = 0.598), but not significant statistically (p = 0.052). Analysis of soil enzyme activities indicated a significant positive correlation between soil TPH concentration and activities of dehydrogenases (r = 0.974, p < 0.001), hydrogen peroxidases (r = 0.957, p < 0.001), polyphenol oxidases (r = 0.886, p < 0.001), and SIR (r = 0.916, p < 0.001). On the contrary, the urease activity showed a negative correlation with paddy soil TPH concentration (r = -0.814, p = 0.002), and could be used as a sensitive indicator of petroleum contamination. PMID- 15888894 TI - Assessment of ambient ozone effects on vegetation using snap bean as a bioindicator species. AB - Tropospheric ozone is an air pollutant that is toxic to plants, causing visible injury to foliage and a reduction in growth and yield. The use of plant bioindicators is one approach to assess the ozone impacts in diverse geographical areas. The objective of this study was to evaluate snap bean (Phaseolus vulgaris L.) as a potential bioindicator species. Three snap bean genotypes known to exhibit a range of ozone sensitivity were grown in pots under charcoal-filtered (CF) or nonfiltered (NF) treatments in open-top chambers, or under ambient air (AA) conditions. Treatment effects on biomass were not significant at 56 days after planting (DAP), but midseason foliar injury increased in the NF and AA treatments relative to CF controls. An increase in ozone from 25 to 30 nL L(-1) in CF controls to approximately 50 nL L(-1) in the NF and AA treatments was found to suppress final pod dry weight per plant by 40 to 60% in the most sensitive genotype S156. The same treatments suppressed final pod dry weight by 20 to 30% in a moderately sensitive genotype Oregon-91, and by 10% or less in a tolerant genotype R123. An S156 to R123 yield ratio of approximately one was observed under CF conditions. The S156 to R123 yield ratio declined to 0.6 to 0.7 in the NF treatment and declined further to 0.4 to 0.5 in the AA treatment, suggesting that ozone impact was underestimated in the open-top chambers. The results suggest that a snap bean bioindicator system has the potential to detect ambient ozone effects at present-day ozone concentrations. PMID- 15888895 TI - Multivariate analysis of paired watershed data to evaluate agricultural best management practice effects on stream water phosphorus. AB - Quantification of the effects of management programs on water quality is critical to agencies responsible for water resource protection. This research documents reductions in stream water phosphorus (P) loads resulting from agricultural best management practices (BMPs) implemented as part of an effort to control eutrophication of Cannonsville Reservoir, a drinking water supply for New York City. Dairy farms in the upstate New York reservoir basin were the target of BMPs designed to reduce P losses. A paired watershed study was established on one of these farms in 1993 to evaluate changes in P loading attributable to implementation of BMPs that included manure management, rotational grazing, and improved infrastructure. Intensive stream water monitoring provided data to calculate P loads from the 160-ha farm watershed for all runoff events during a two-year pre-treatment period and a four-year post-treatment period. Statistical control for inter-annual climatic variability was provided by matched P loads from a nearby 86-ha forested watershed, and by several event flow variables measured at the farm. A sophisticated multivariate analysis of covariance (ANCOVA) provided estimates of both seasonal and overall load reductions. Statistical power and the minimum detectable treatment effect (MDTE) were also calculated. The results demonstrated overall event load reductions of 43% for total dissolved phosphorus (TDP) and 29% for particulate phosphorus (PP). Changes in farm management practices and physical infrastructure clearly produced decreases in event P losses measurable at the small watershed scale. PMID- 15888896 TI - Tillage and nutrient source effects on water quality and corn grain yield from a flat landscape. AB - Beneficial effects of leaving residue at the soil surface are well documented for steep lands, but not for flat lands that are drained with surface inlets and tile lines. This study quantified the effects of tillage and nutrient source on tile line and surface inlet water quality under continuous corn (Zea mays L.) from relatively flat lands (<3%). Tillage treatments were either fall chisel or moldboard plow. Nutrient sources were either fall injected liquid hog manure or spring incorporated urea. The experiment was on a Webster-Canisteo clay loam (Typic Endoaquolls) at Lamberton, MN. Surface inlet runoff was analyzed for flow, total solids, NO(3)-N, NH(4)-N, dissolved P, and total P. Tile line effluent was analyzed for flow, NO(3)-N, and NH(4)-N. In four years of rainstorm and snowmelt events there were few significant differences (p < 0.10) in water quality of surface inlet or tile drainage between treatments. Residue cover minimally reduced soil erosion during both snowmelt and rainfall runoff events. There was a slight reduction in mineral N losses via surface inlets from manure treatments. There was also a slight decrease (p = 0.025) in corn grain yield from chisel-plow plots (9.7 Mg ha(-1)) compared with moldboard-plow plots (10.1 Mg ha(-1)). Chisel plowing (approximately 30% residue cover) alone is not sufficient to reduce nonpoint source sediment pollution from these poorly drained flat lands to the extent (40% reduction) desired by regulatory agencies. PMID- 15888897 TI - A method for determining the phosphorus sorption capacity and amorphous aluminum of aluminum-based drinking water treatment residuals. AB - A high amorphous aluminum or iron oxide content in drinking water treatment residuals (WTRs) can result in a high phosphorus (P) sorption capacity. Therefore, WTR may be used beneficially to adsorb P and reduce P loss to surface or ground water. The strong relationship between acid ammonium oxalate extractable aluminum (Al(ox)) and Langmuir phosphorus adsorption maximum (P(max)) in WTR could provide a useful tool for determining P(max) without the onus of the multipoint batch equilibrations necessary for the Langmuir model. The objectives of this study were to evaluate and/or modify an acid ammonium oxalate extraction of Al(ox) and the experimental conditions used to generate P adsorption isotherms to strengthen the relationship between Al(ox) and P(max). The oxalate extraction solution to WTR ratio varied from 40:1, 100:1, and 200:1. Batch equilibration conditions were also varied. The WTR particle size was reduced from <2 mm to <150 microm, and batch equilibration was extended from 17 h to 6 d. Increasing the solution to WTR ratio to 100:1 extracted significantly greater Al(ox) at levels of >50 mg Al kg(-1). No additional increase was found at 200:1. Reducing WTR particle size from <2 mm to <150 microm increased P(max) 2.46-fold. Extending the equilibration time from 17 h to 6 d increased P(max) by a mean of 5.83-fold. The resulting empirical regression equation between the optimized Al(ox) and P(max) (r(2) = 0.91, significant at the 0.001 probability level) may provide a tool to estimate the P(max) of Al-based WTR simply by measuring Al(ox). The accurate determination of WTR P(max) and Al(ox) is essential in using WTR effectively to reduce P loss in runoff or to reduce the solubility of P in agricultural soils or organic waste materials (biosolids, manure). PMID- 15888898 TI - Management strategy impacts on ammonia volatilization from swine manure. AB - Ammonia emitted from manure can have detrimental effects on health, environmental quality, and fertilizer value. The objective of this study was to measure the potential for reduction in ammonia volatilization from swine (Sus scrofa domestica) manure by temperature control, stirring, addition of nitrogen binder (Mohave yucca, Yucca schidigera Roezl ex Ortgies) or urease inhibitor [N-(n butyl) thiophosphoric triamide (NBPT)], segregation of urine from feces, and pH modification. Swine manure [total solids (TS) = 7.6-11.2%, total Kjeldahl nitrogen (TKN) = 3.3-6.2 g/L, ammonium nitrogen NH(+)(4)-N = 1.0-3.3 g/L] was stored for 24, 48, 72, or 96 h in 2-L polyvinyl chloride vessels. The manure was analyzed to determine pre- and post-storage concentrations of TS and volatile solids (VS), TKN, and NH(+)(4)-N. The concentration of accumulated ammonia N in the vessel headspace (HSAN), post-storage, was measured using grab sample tubes. Headspace NH(3) concentrations were reduced 99.3% by segregation of urine from feces (P < 0.0001). Stirring and NBPT (152 microL/L) increased HSAN concentration (119 and 140%, respectively). Headspace NH(3) concentration increased by 2.7 mg/m(3) for every 1 degree C increase in temperature over 35 degrees C. Slurry NH(+)(4)-N concentrations were reduced by segregation (78.3%) and acidification to pH 5.3 (9.4%), and increased with stirring (4.8%) and increasing temperature (0.06 g/L per 1 degree C increase in temperature over 35 degrees C). Temperature control, urine-feces segregation, and acidification of swine manure are strategies with potential to reduce or slow NH(+)(4)-N formation and NH(3) volatilization. PMID- 15888899 TI - Proton binding by humic and fulvic acids from pig slurry and amended soils. AB - The knowledge of acid-base characteristics of humic acid (HA) and fulvic acid (FA) fractions of organic amendments and amended soils is of considerable importance for assessing their agronomic efficacy and environmental impact. In this work, the acid-base properties of HAs and FAs isolated from pig slurry, soils amended with either 90 or 150 m(3) ha(-1) yr(-1) of pig slurry for 3 yr, and the corresponding nonamended control soil were investigated by using a current potentiometric titration method. The nonideal competitive adsorption (NICA) model that describes proton binding by two classes of binding sites (carboxylic- and phenolic-type groups) was successfully fit to titration data. With respect to the control soil HA and FA, pig-slurry HA and FA were generally characterized by smaller carboxylic-type group contents, slightly smaller phenolic-type group contents, larger affinities for proton binding by the carboxylic-type groups, and much smaller, in the case of the HA fraction, or similar, in the case of the FA fraction, affinities for proton binding by the phenolic-type groups. Amendment with pig slurry determined a number of modifications in soil HAs and FAs, including decrease of acidic functional group contents, and slight increase of the proton affinity of the carboxylic-type groups. Further, a slight decrease of the affinities for proton binding by the phenolic-type groups of HAs was observed. These effects can have a large impact on the biological availability, mobilization, and transport of macro- and micronutrients, toxic metal ions, and xenobiotic organic cations in pig slurry amended soils. PMID- 15888900 TI - Postoperative care following pituitary surgery. AB - Patients undergoing surgery for pituitary tumors represent a heterogeneous population each with unique clinical, biochemical, radiologic, pathologic, neurologic, and/or ophthalmologic considerations. The postoperative management of patients following pituitary surgery often occurs in the context of a dynamic state of the hypothalamic-pituitary-end organ axis. Consequently, a significant component of the postoperative care of these patients focuses on vigilant screening and observation for neuroendocrinologic perturbations such as varying degrees of hypopituitarism and disorders of water balance (diabetes insipidus and the syndrome of inappropriate antidiuretic hormone). Additionally, one must be cognizant of other potential complications specific to the transsphenoidal approach for tumor removal including cerebrospinal fluid leakage and meningitis. This review addresses the postoperative management of patients undergoing pituitary surgery with an emphasis on careful screening and recognition of complications. PMID- 15888901 TI - Role of the interventional pulmonologist in the intensive care unit. AB - Interventional pulmonology is a new field within the pulmonary and critical care medicine specialty with a focus on invasive diagnostic and therapeutic modalities in airway and pleural disorders. The interventional pulmonologist is highly qualified to take a prominent role in the intensive care unit in a consultative fashion to provide assistance with pleural procedures, establishment and care of artificial airways, and management of patients with respiratory failure attributable to structural central airway disorders. The presence of a dedicated operator with advanced skills facilitates access to specialized procedures in an expeditious and safe manner. Clear communication between the interventional pulmonologist and intensivist is vital to ensure a collaborative effort that delivers optimal patient care. PMID- 15888902 TI - Outcome of morbid obesity in the intensive care unit. AB - This was a retrospective chart review of consecutive obese patients admitted to the medical intensive care unit. Patients were divided into 2 groups: mild to moderately obese (group 1, body mass index =30-40 kg/m(2)) and morbidly obese (group 2, body mass index >40 kg/m(2)). Acute Physiology and Chronic Health Evaluation II scores were not significantly different between the 2 groups. Morbidly obese patients (group 2) had higher rates of mortality and nursing home admission. They also showed higher rates of intensive care unit complications including sepsis, nosocomial pneumonia, acute respiratory distress syndrome, catheter infection, tracheostomy, and acute renal failure. Their median length of mechanical ventilation was longer (2 days, range 2-12 vs 9 days, range 1-37,P = .009). In a logistic regression analysis, morbid obesity remained a significant predictor of death or disposition to nursing home even after controlling for age (P = .019, odds ratio = 7.60, 95% confidence interval = 1.39-41.6). Morbidly obese patients (body mass index >40 kg/m(2)) admitted to intensive care units have higher rates of mortality, nursing home admission, and intensive care unit complications and have longer stays in the intensive care unit and time on mechanical ventilation. PMID- 15888903 TI - Refeeding syndrome and hypophosphatemia. AB - Hypophosphatemia is a potentially life-threatening complication of reinstating nutrition in a malnourished patient. Refeeding syndrome is a term that refers to various metabolic abnormalities that may complicate carbohydrate administration in subnourished patient populations. Hypophosphatemia is the most well-known, and perhaps most significant, element of the refeeding syndrome and may result in sudden death, rhabdomyolysis, red cell dysfunction, and respiratory insufficiency. This review briefly examines refeeding-induced hypophosphatemia in the hospitalized patient in hopes of making clinicians more aware of this common, but often overlooked, potentially dangerous problem. PMID- 15888904 TI - Fatal rapidly progressive interstitial pneumonitis associated with amyopathic dermatomyositis and CD8 T lymphocytes. AB - A patient with amyopathic dermatomyositis associated with fatal rapidly progressive interstitial pneumonitis resistant to therapy is described. Pathologic examination of a transbronchial lung biopsy specimen showed diffuse alveolar damage and nonspecific interstitial pneumonia-organizing pneumonia-like findings. Bronchoalveolar lavage fluid contained many CD8+ lymphocytes, considered to be cytotoxic T cells. Analysis of bronchoalveolar lavage fluid in this case may provide prognostically and pathogenetically important information. PMID- 15888905 TI - Management of delirium tremens. AB - Delirium tremens is recognized as a potentially fatal and debilitating complication of ethanol withdrawal. Research thus far has primarily focused on the prevention of delirium tremens. PMID- 15888906 TI - Refeeding syndrome and hypophosphatemia. PMID- 15888907 TI - Creutzfeldt-Jakob disease presenting as refractory nonconvulsive status epilepticus. PMID- 15888909 TI - Role of formyl peptide receptor-like 1 (FPRL1/FPR2) in mononuclear phagocyte responses in Alzheimer disease. AB - Alzheimer disease (AD) is the most common neurodegenerative disease, affecting approx 4 million people in the United States in the year 2000 alone. Amyloid beta (Abeta) deposition, activated glial cells, and neuritic degeneration are the characteristic features of AD. Although the precise cause of AD has yet to be determined, a bulk of evidence suggests that inflammatory responses elicited by elevated Abeta peptides play an important role in the pathogenic process of the disease. In AD brain, mononuclear phagocytes (microglia) accumulate at the sites of Abeta peptide deposition. In vitro, Abeta peptides activate mononuclear phagocytes to release neurotoxic mediators. A number of cell-surface molecules have been reported to act as putative receptors for Abeta peptides, among which the G protein-coupled formyl peptide receptor-like 1 (FPRL1) and its mouse homolog FPR2 have been shown to be expressed by activated microglial cells and mediate the chemotactic activity of the 42 amino acid form of Abeta (Abeta42). FPRL1 also participates in Abeta42 internalization in macrophages and its cytotoxicity for neuronal cells. Therefore, FPRL1 may be involved in the inflammatory aspects of AD. This review discusses recent findings relevant to the function and regulation of FPRL1/FPR2 in mononuclear phagocytes by pro- and antiinflammatory signals and its potential as a therapeutic target in AD. PMID- 15888910 TI - Role of regulatory invariant CD1d-restricted natural killer T-cells in protection against type 1 diabetes. AB - Invariant CD1d-restricted natural killer T (iNKT) cells function during innate and adaptive immune responses. A functional and numerical deficiency of iNKT cells is well documented in both nonobese diabetic (NOD) mice and humans with autoimmune type 1 diabetes (T1D). Restoring the numerical and/or functional deficiency of iNKT cells in NOD mice by either treatment with alpha galactosylceramide, transgenic induction of Valpha14-Jalpha18 expression, or transgenic expression of CD1d in NOD islets under the control of the human insulin promoter confers protection from T1D in these mice. Recently, considerable progress has been made in understanding the developmental and functional activities of iNKT cells. In this review, we discuss the role of iNKT cell deficiency and defective development in the onset of T1D in NOD mice and the different protective mechanisms known to restore these defects. PMID- 15888911 TI - Immunologic roles of hyaluronan. AB - Hyaluronan (HA), a large glycosaminoglycan composed of D-N-acetylglucosamine and D-glucuronic acid, is expressed in virtually all tissues and has long been considered to serve as a structural component or filling material in the tissue interstitium (Filler Theory). This idea was revised with the discovery of HA binding proteins that introduced the concept that HA may also serve as an adhesive substrate for cellular trafficking (Adhesion Theory). Most recently, it has been shown that HA fragments can deliver maturational signals to dendritic cells (DCs) and high molecular weight HA polymers can deliver costimulatory signals to T-cells (Signaling Theory). Thus, HA may represent an important component of the immune system. Recently, we have evaluated the impact of HA on Langerhans cell (LC) maturation and migration using a novel peptide inhibitor of HA function, termed Pep-1 (GAHWQFNALTVR). As skin-specific members of the DC family, LCs are crucial for the initiation of cutaneous immune responses. Local injections of Pep-1 prevented hapten-induced LC migration from the epidermis, providing the first experimental evidence that HA facilitates their emigration. Moreover, Pep-1 also significantly inhibited the hapten-induced maturation of LCs in vivo as assessed by cell morphology, costimulatory molecule expression, and their ability to induce proliferation of allogeneic T-cells. HA therefore has dual functionality to facilitate LC migration and maturation, the two critical events for the initiation of adaptive immune responses. Finally, we have observed that DC-dependent, antigen-specific T-cell proliferation and cytokine secretion is blocked by Pep-1. These results have revealed a previously unrecognized role for HA in antigen presentation. Thus, far from an inert structural biopolymer, HA represents a multifunctional carbohydrate mediator of immune processes. PMID- 15888912 TI - Threat matrix: low-molecular-weight hyaluronan (HA) as a danger signal. AB - Whether or not T cell receptor engagement leads to full activation or tolerance is determined by the context in which the antigen is encountered. Antigen presented by activated APCs in the presence of costimulation leads to full T cell activation, while antigen presented by resting APCs leads to tolerance. Pathogen associated molecular patterns in the form of toll-like receptor ligands play a critical role in activating APCs and promoting T cell activation. In this review we hypothesize that low-molecular-weight species of the extracellular matrix polymer hyaluronan also performs this function by acting as an endogenous danger signal. PMID- 15888913 TI - Discoidin domain receptor 1: a new class of receptor regulating leukocyte collagen interaction. AB - Previous studies demonstrated that type I collagen, a major component of the extracellular matrix, could influence the differentiation and function of leukocytes; however, it is not clear whether those effects of collagen were based on its interaction with the classic collagen receptors, alpha1beta1 and alpha2beta1 integrins. We recently detected significant upregulation of discoidin domain receptor 1 (DDR1), a new class of collagen receptor, in human leukocytes, including neutrophils, monocytes, and lymphocytes, in vitro, leading to the hypothesis that the leukocyte-activating effects of collagen might be owing to its interaction with DDR1. In this review, we summarize our recent findings demonstrating that DDR1-collagen interaction facilitates the adhesion, migration, differentiation/maturation, and cytokine/chemokine production of leukocytes. We also describe the intracellular signaling pathways activated by DDR1 interaction with collagen. PMID- 15888914 TI - STAT4: a critical regulator of inflammation in vivo. AB - Signal transducer and activator of transcription 4 (STAT4) is a central mediator in generating inflammation during protective immune responses and immune-mediated diseases. In the 8 yr since their first description, STAT4-deficient mice have defined the role of STAT4 in a variety of in vivo model systems. Despite the extensive study and use of these mice, the exact role of STAT4 in vivo is still unclear. In this review, I focus on describing the phenotypes of STAT4-deficient immune responses to pathogens and in diseases. Comparing the effects of STAT4 deficiency among numerous model systems will further enhance the development of a systemic model of STAT4 function in vivo. PMID- 15888915 TI - The proteasome: a central regulator of inflammation and macrophage function. AB - Proteasomes, multisubunit complexes that consist of a 20S proteasome and a 19S regulatory complex, are essential for several cellular processes. Our interest in the proteasome complex stems from our observations that a novel photoactivable lipopolysaccharide (LPS) probe binds to specific proteasome subunits, and that LPS enhances the chymotrypsin-like activity of the proteasome to degrade synthetic peptides in vitro. Experiments with proteasome inhibitors have shown that expression of many LPS-inducible genes, including TLR2, is inhibited in macrophages. More important, proteasome inhibitors such as lactacystin can prevent LPS-induced shock in mice. This article focuses on the role of the proteasome in the development of inflammatory processes, which may result in septic shock, hemorrhagic shock, atherosclerosis, and neurodegenerative disorders. Taken collectively, the results suggest a potentially important role of the proteasome in inflammation and other macrophage functions. PMID- 15888916 TI - Interaction between HSP73 and HLA-DRB1*0401: implications for the development of rheumatoid arthritis. AB - The amino acid motif QKRAA on HLA-DRB1*0401 carries susceptibility to develop rheumatoid arthritis through unknown mechanisms. We identified the original functions of this motif. In B-cells, HSP73, the constitutive 70-kDa heat-shock protein (HSP), associates with HLA-DRB1*0401. This interaction causes abnormal trafficking of HLA-DRB1*0401. Indeed, HSP73 targets HLA-DRB1*0401 from endoplasmic reticulum to lysosomes bypassing the normal route through the Golgi apparatus and endosomes. In this article, we propose mechanisms to explain how 70 kDa HSPs might contribute to rheumatoid arthritis. PMID- 15888917 TI - Focal adhesion kinase-related protein tyrosine kinase Pyk2 in T-cell activation and function. AB - Pyk2 is a protein tyrosine kinase expressed primarily in brain and hematopoietic cells. It becomes activated in response to stimulation through numerous receptors, including integrins, chemokine receptors, and antigen receptors, and is found in association with src-family kinases. Although this enzyme associates with many proteins known to be important for activation and has many characteristics of a scaffolding protein, its function remains elusive. A number of studies in non-T-cells suggest that Pyk2 is important for cell spreading, cell migration, and integrin function; however, a defined role in T-cells has not been established. Here, we discuss evidence that implicates Pyk2 in directionality of signaling, which is essential to establishment of the directional killing mediated by cytotoxic lymphocytes. PMID- 15888918 TI - Hypopituitarism as a late complication of hemorrhagic fever. AB - We report three patients who developed hypopituitarism as a late complication of hemorrhagic fever with renal syndrome (HFRS). Their past history, physical examination, and endocrine investigation confirmed hypopituitarism. Magnetic resonance imaging of the pituitary revealed atrophic pituitary gland with an empty sella. Hemorrhagic fever is endemic in certain regions of the Balkans, and this preliminary report suggests the importance of investigating the endocrine status in every patient who survived HFRS. PMID- 15888919 TI - Sarcosine1, glycine8 angiotensin II is a functional AT1 angiotensin receptor antagonist. AB - Sarcosine1, glycine8 angiotensin II (SG Ang II) displayed unusual characteristics in early pharmacological studies. It was a potent antagonist of the dipsogenic actions of intracerebroventricularly administered Ang II in the rat, but showed low affinity for bovine cerebellar Ang II receptors. It has recently been shown that SG Ang II binds preferentially to the AT1 receptor. To determine if SG Ang II is a functional antagonist of the AT1 receptor-mediated calcium signaling, CHO cells stably transfected with AT1 receptors were exposed to Ang II in the presence and absence of SG Ang II. At concentrations of 10-100 nM, SG Ang II completely inhibited Ang II-stimulated intracellular Ca2+ mobilization in AT1A and AT1B receptor-transfected cells. SG Ang II and 125I- SG Ang II bound to AT1A and AT1B receptor-transfected cells with KD and KI values of 2-4 nM. In contrast, SG Ang II bound to AT2 transfected cells with a KI value of 7.86 microM. These results demonstrate that SG Ang II is a selective and functional peptide antagonist of the AT1 angiotensin II receptor subtype. PMID- 15888920 TI - Gonadal steroid modulation of the limbic-hypothalamic- pituitary-adrenal (LHPA) axis is influenced by social status in female rhesus monkeys. AB - Chronic stress can have a deleterious effect on the re-productive axis that, for females, is manifested in an increased incidence of infertility. However, gonadal steroids may, in turn, affect a female's response to stress as measured by activity within the limbic-hypothalamic-pituitary-adrenal (LHPA) axis. What is not clear is whether a history of exposure to stress modifies the effect of gonadal steroids on LHPA responsivity. Rhesus monkeys present a unique opportunity to assess LHPA responsivity when housed socially in groups. Under these situations, monkeys exhibit a rich network of affiliation and have established social status hierarchies. Previous work indicates that socially subordinate macaque females are hypercortisolemic due to diminished gluco corticoid negative feedback. The present study tested the hypothesis that estradiol (E2) would decrease gluco-corticoid negative feedback, assessed from a dexamethasone (DEX) suppression test, and increase the response to corticotropin releasing factor (CRF) and that these effects would be attenuated by co-treatment with P4. In addition, we also determined whether E2 and P4 would differentially affect LHPA responsiveness to pharmacological challenge in socially dominant compared with subordinate females. Endogenous gonadal hormone secretion in female rhesus monkeys (n = 7) was suppressed by continuous treatment with a sustained release formulation of the GnRH analog leuprolide acetate (Lupron Depot). The response to a combined DEX suppression-CRF stimulation test was assessed using a counterbalanced design during a placebo (control) treatment condition and during E2, P4, and E2 + P4 re-placement therapy. Females who were members of a large breeding group of 140 adults and juveniles of both sexes, were classified as dominant (n = 4) or subordinate (n = 3) based on the relative social dominance positions within the group. Plasma levels of cortisol were significantly higher during E2 replacement compared to the other treatment conditions following DEX suppression and stimulation with CRF. PMID- 15888921 TI - Orexin a stimulates hypothalamic-pituitary-adrenal (HPA) axis function, but not food intake, in the absence of full hypothalamic NPY-ergic activity. AB - Neonatal monosodium L-glutamate (MSG) treatment destroys hypothalamic arcuate nucleus neuronal bodies, thus inducing several metabolic abnormalities. As a result, rats develop a phenotype characterized by hyperleptinemia and by impaired NPY but normal preproorexin hypothalamic mRNAs expression. Thus, our study was designed to explore whether hypothalamic effects of orexin A on food intake and glucocorticoid production develop in the absence of full hypothalamic NPY-ergic activity. For this purpose we evaluated, in control and MSG-treated rats, the consequences of intracerebroventricular (icv) orexin A administration on food intake and changes in circulating levels of ACTH and glucocorticoid. Our results indicate that orexin A icv treatment stimulated hypothalamic-pituitary-adrenal (HPA) axis activity in both MSG-damaged and normal animals, with this response even more pronounced in neurotoxin-damaged rats. Conversely, food intake was only enhanced by icv orexin A injection in normal rats. Our study further supports that acute hypothalamic effects of orexin A on food intake and glucocorticoid production are due to independent neuronal systems. While intact arcuate nucleus activity is needed for the orexinergic effect induced by icv orexin A administration, conversely, orexin A-stimulated HPA axis function takes place even in the absence of full NPY-ergic activity. PMID- 15888922 TI - Effects of an olive oil-enriched diet on plasma GLP-1 concentration and intestinal content, plasma insulin concentration, and glucose tolerance in normal rats. AB - The present study aims mainly at measuring, in normal rats, the GLP-1 response to oral intake of an olive oil-enriched diet (OO), and at assessing the long-term effects of such a diet on the GLP-1 content of the intestinal tract, as well as the plasma D-glucose, insulin, and GLP-1 pattern during an oral glucose tolerance test. In meal-trained rats, the mean increment in plasma GLP-1 concentration at min 10 and 20 was 1.39 +/- 0.23 ng/mL higher (p < 0.001) in the rats given access to the OO diet rather than control diet. Relative to the initial value (d 0), the gain in body weight at d 50 was also higher in the animals fed the OO rather than control diet. At d 50, the GLP-1 content of the jejunum, ileum, colon, and cecum were not significantly different in the two groups of rats. At d 19 and 36, the increment in both plasma insulin concentration and paired ratio between plasma insulin and D-glucose concentrations were again higher, during an oral glucose tolerance test conducted in overnight fasted animals, in the rats otherwise fed the OO, as distinct from control, diet. The intake of an olive oil-enriched diet thus increases, in normal rats, GLP-1 release, this coinciding during long-term exposure to the OO diet with higher body weight gain, increased secretory response of insulin-producing cells to oral glucose administration, and, after 36 d, improved glucose tolerance. PMID- 15888923 TI - Igf-I and postnatal growth of weaver mutant mice. AB - IGF-I is an anabolic growth factor essential for growth and development, both as a mediator of growth hormone (GH) action and as a local stimulator of cell proliferation and differentiation. Although the importance of IGF-I in postnatal growth has been studied for several decades, its functions in pathological states are not fully understood. The weaver (wv) mutant mouse is a commonly used model for studying hereditary cerebellar ataxia and provides us with an opportunity to study the function of IGF-I in postnatal growth during neurodegeneration. In prepubertal wv mice, we found a parallel decrease in body weight and serum IGF-I. This parallel relationship was maintained in females, but not in males, as wv mice entered puberty. Interestingly, we found an increase in the levels of circulating IGF-I and hepatic mRNA preceded the catch-up of body weight of pubertal male wv mice. The increase in IGF-I levels coincided with a surge of circulating androgen at the onset of male puberty, suggesting that androgen might trigger the increase in IGF-I production in the pubertal and adult male wv mice. Overall, our results support the concept that IGF-I plays an important role in postnatal growth during and after neurodegeneration of wv mice. In addition, IGF I's regulation of systemic growth during and after puberty is likely modulated by androgen in male wv mice. PMID- 15888924 TI - Views from within and beyond: narratives of cardiac contractile dysfunction under senescence. AB - Senescence is associated with enhanced risk of cardio-vascular diseases. It is generally considered that decline in growth hormones (such as insulin-like growth factor I), intrinsic myocardial and endothelial functions, as well as accumulation of reactive oxygen species with increased age may contribute to cardiovascular senescence. It is believed that heart function, especially cardiac reserve declines with advanced age. However, most experimental and clinical investigations on ventricular function only included young or adult subjects and failed to address this important age issue in heart pathophysiology. Although senescent but otherwise healthy hearts may possess normal pumping function at the resting or non-stressed state, some aging-associated factors such as accumulation of reactive oxygen species and activation of selective stress signaling path-ways may interact with certain risk factors and compromise overall cardiac function. The precise cause and progression of compromised cardiac function in the elderly remain controversial. This review will focus on senescence-related alterations in cardiac contractile function with a special emphasis on oxidative stress and activation of stress signaling. PMID- 15888925 TI - Suppression of body weight gain preserves acute insulin response to glucose in the portal vein of spontaneously type 2 diabetic rats with visceral obesity. AB - The age-related changes in acute insulin response after glucose loading and the influence of suppression of body weight gain were investigated by using blood samples from portal and peripheral veins. We placed indwelling catheters in the portal vein of 12- and 24- wk-old Otsuka Long-Evans Tokushima fatty (OLETF) rats (n = 8, 12), and age-matched control Long-Evans Tokushima Otsuka (LETO) rats (n = 8, 6). To suppress the body weight gain, 6 out of 12 OLETF rats were fed chow containing 50 ppm voglibose (VOG) from 8 until 24 wk of age. After fasting for 17 h, rats underwent 1 g/kg oral glucose tolerance test (OGTT). Peripheral glucose levels after glucose loading were significantly higher in 12- and 24-wk-old OLETF rats than in the age-matched LETO rats. Values for delta insulin 15 min/delta glucose 15 min (delta I15 min/delta G15 min) in portal blood were 0.029 +/- 0.011 and 0.009 +/- 0.009 (12 wk of age) and 0.03 +/- 0.03 and -0.01 +/- 0.01 (24 wk of age) in the LETO rats and OLETF rats. At the age of 24 wk, the body weights in VOG-treated OLETF rats were significantly lower than those in the OLETF rats. And there was significantly greater acute insulin response to glucose in VOG-treated OLETF rats than in the OLETF rats. Acute insulin response to glucose decreased with advancing age and the suppression of body weight gain preserved the response in spontaneously type 2 diabetic rats with visceral fat obesity. PMID- 15888926 TI - Neural activity between ovaries and the prevertebral celiac-superior mesenteric ganglia varies during the estrous cycle of the rat. AB - The ovaries' innervation arrives via the superior ovarian nerve, which originates from the celiac ganglion. Using True Blue as an antidromic marker, the present study analyzed the changes in the anatomical relation between each ovary and the prevertebral celiac-superior mesenteric ganglia during the estrous cycle. The number of labeled neurons increased from the day of diestrus 1 to the day of proestrus. The largest number of labeled cells was observed when tracer was injected into the left ovary on proestrus. The number of labeled cells was significantly higher when the tracer was injected into the left ovary on proestrus than when it was done in the right one. When tracer was injected into the left ovary, the average labeled area of cells increased significantly from diestrus 1 to proestrus, and declined at estrus. In contrast, when True Blue was injected into the right ovary, the average labeled area was similar in diestrus 1 and diestrus 2, and the values increased in proestrus and estrus. The results indicate an apparent asymmetry in the activity of neural connections between ovaries and the prevertebral celiac-superior mesenteric ganglia, and that the number of active neurons of these connections varies during the estrous cycle. PMID- 15888927 TI - Induction of uterine prostaglandin H synthase 2 by estradiol following fetal adrenalectomy. AB - In sheep, fetal cortisol stimulates the conversion of progesterone to estradiol in late gestation initiating labor. It is unclear whether an intact fetal hypothalamic-pituitary-adrenal (HPA) axis is required to induce the estradiol triggered subsequent endocrine changes including enhanced intrauterine prostaglandin (PG) synthesis associated with the onset of labor. We have shown that maternal estradiol administration stimulates PG H synthase (PGHS)-2 expressions in pregnant ovine intra-uterine tissues. The current study was undertaken to determine whether the fetal adrenal mediates estradiol's stimulation of the intrauterine PGHS-2 in pregnant sheep. Placenta, myometrium, and endometrium were collected from two groups of ewes at 123-127 d of gestational age (dGA) after fetal adrenalectomy and vehicle treatment (ADX; n = 5); or fetal ADX and maternal estradiol administration (5 mg twice a day for 2 d, ADX+E2, n = 5). PGHS-2 mRNA and protein were analyzed by Northern and Western Blot analyses in both groups and presented as the ratios to beta actin mRNA for Northern and G protein beta subunit for Western blot analysis. Fetal plasma cortisol was measured by radioimmunoassay. Data were analyzed by Student's t test. Fetal plasma cortisol levels were low in ADX and ADX+ E2 groups (<6 ng/mL). The cervix of all ADX+E2 treated ewes was dilated at necropsy. Three out of five ADX+ E2-treated ewes delivered within 48 h. The cervix was closed in all fetal ADX ewes at necropsy. PGHS-2 mRNA and protein increased (p < 0.05) in myometrium and endometrium, but not placenta in ADX+E2-treated ewes compared with ADX group. These data provide the first in vivo evidence for estradiol upregulation of intrauterine PGHS-2 in late gestation in the absence of an intact fetal HPA axis. Thus, the fetal adrenal is not required to mediate estradiol's stimulation of uterine PGHS-2 expression associated with the onset of labor. PMID- 15888928 TI - Neonatally administered tert-octylphenol affects onset of puberty and reproductive development in female rats. AB - There now is evidence that many of the synthetic chemicals released into the environment can impact on the function of the endocrine system of many organisms. One group of chemicals, the alkylphenols, used in paints, pesticides, herbicides, detergents, and plastics, has been found to have the ability to bind estrogen receptors. This estrogenic property makes these compounds potentially hazardous to the developing reproductive system and neuroendocrine brain. In this study we deter- mined the effects of exposure to the environmental toxins 4-nonylphenol (NP) and 4-tert-octylphenol (OP) and to synthetic estrogen diethylstilbesterol (DES) during the early postnatal period (d 0-10) on the development of reproductive function. The day of vaginal opening, ovulation, prepubertal LH levels, LH response to estradiol, estrous cyclicity, and ovarian histology were determined. In the OP- and DES-treated groups, the vaginal opening was observed to have occurred several days prior to that of the control group. The NP-treated group showed vaginal opening at ages similar to those of the control group. Treatment with OP prevented ovulation in a significant number of animals, as well as in all animals treated with DES, whereas the control and NP-treated animals ovulated normally. Animals treated with DES and OP had significantly lower ovarian weights and higher uterine weights than either control animals or NP treated animals. Higher basal LH levels, as well as the absence of the prepubertal LH surge, were observed in both DES- and OP-treated animals. A significant number of OP-treated animals showed no LH response to the estradiol 17beta challenge. NP-treated animals responded positively to the estradiol-17beta challenge. Persistent estrus was also apparent in both OP- and DES-treated animals. Upon histological examination, the ovaries in OP-treated animals were found to have a decreased number of corpora lutea and an increased number of preantral and atretic follicles. These data suggest that exposure to OP during the critical period of sexual brain differentiation affects the onset of puberty and reproductive development. PMID- 15888929 TI - Alterations in reproductive function in SRC tyrosine kinase knockout mice. AB - The role of Src tyrosine kinase in regulating reproductive processes in female mice was investigated using Src wild-type (+/+), heterozygous (+/-), and knockout (-/-) mice. Ovarian Src kinase activity transiently increased in Src +/+ mice following eCG administration. Src knockout mice were infertile. Estrous cycles and vaginal opening in Src knockouts were variable and altered compared with Src +/+ and +/- mice. Follicle development was compromised in Src knockout mice as evidenced by reduced numbers of large pre-antral and antral follicles compared to Src +/+ mice. Corpora lutea were not observed in the ovaries of Src knockout mice; however, administration of eCG and hCG did result in ovulation. Serum LH and FSH on d 40 and 52 of age did not differ between Src wild-type and knockout females. Results from these studies reveal that female Src knockout mice are infertile due to reduced follicle development and anovulation. PMID- 15888930 TI - Susceptibility testing methods of antifungal agents. AB - Several methods for testing antifungal susceptibility are currently utilized. Minimum inhibitory concentrations can be tested using standardized noncommercial or commercial tests. Fungicidal testing includes either in vitro methods, such as time-kill or minimum fungicidal testing methods, or animal models. This chapter provides background information for utilizing and evaluating results obtained from antifungal susceptibility testing methods. PMID- 15888931 TI - Deoxyribonucleic acid fingerprinting methods for Candida species. AB - A number of genetic fingerprinting methods have evolved to analyze the population structure and to perform epidemiological and etiological studies of infectious fungi. These methods include multilocus enzyme electrophoresis, restriction fragment-length polymorphism using complex probes, random amplification of polymorphic DNA, and multilocus sequence typing, which are described in this chapter. PMID- 15888932 TI - CARE-2 fingerprinting of Candida albicans isolates. AB - The emergence of resistance to antifungal drugs in medically important fungi has become a significant problem in recent years. Probably the best-studied example is the development of resistance to the widely used antifungal agent fluconazole in the yeast Candida albicans. The availability of matched series of clinical isolates representing the same strain in which drug resistance developed over time has provided opportunities to detect cellular alterations that are correlated with drug resistance. We describe a method for DNA fingerprinting of C. albicans isolates based on Southern hybridization of genomic DNA with the C. albicans-specific repetitive DNA element CARE-2. Molecular typing with CARE-2 permits highly reliable discrimination of unrelated strains to ascertain that serial isolates recovered from individual patients indeed represent the same C. albicans strain. PMID- 15888933 TI - Targeted gene deletion in Candida albicans wild-type strains by MPAR flipping. AB - Many genetic and phenotypic changes occur during the development of drug resistance in fungi. A straightforward approach to assess the contribution of a specific gene to drug resistance is to examine its inactivation in a resistant isolate and to analyze the effect of the mutation on the resistance phenotype. The generation of knockout mutants in the diploid yeast Candida albicans requires two rounds of gene replacement to inactivate both alleles of a target gene. Because auxotrophic markers are not useful for the genetic manipulation of wild type, clinical isolates, dominant selection markers are required. In this chapter, we describe the MPAR flipping method that combines dominant selection with recombinase-mediated marker recycling for targeted inactivation of specific genes in C. albicans wild-type strains. Using the MPAR flipper makes drug resistant clinical isolates amenable to genetic manipulation, a prerequisite for the study of causal relationships between specific genes and drug resistance. PMID- 15888934 TI - Application of deoxyribonucleic acid microarray analysis to the study of azole antifungal resistance in Candida albicans. AB - The near completion of sequencing the Candida albicans genome has made it possible to employ genomic technologies, such as microarray analysis, to aid in identifying key genes involved in such clinical problems as the acquisition of high-level resistance to azole antifungal agents. Here, we outline in detail the methodologies utilized in our laboratory to culture clinical isolates of C. albicans, isolate ribonucleic acid from such cultures, synthesize labeled complimentary deoxyribonucleic acid probes from the ribonucleic acid samples, hybridize the probes to microarray chips, analyze the data from such hybridizations, and validate results using reverse transcriptase-polymerase chain reactions. Microarray analysis gives researchers the ability to identify genes involved in processes such as acquisition of azole resistance and to use the data in a way that may lead to clinical approaches to inactivate these genes and improve patient outcomes. PMID- 15888935 TI - Application of proteomic analysis to the study of azole antifungal resistance in Candida albicans. AB - The sequencing of the Candida albicans genome and recent refinements in protein resolution and identification techniques have greatly enhanced the application of proteomics for the study of this fungal pathogen. Proteome analysis includes the separation and isolation of proteins by two-dimensional polyacrylamide gel electrophoresis and subsequent protein identification by peptide mass fingerprinting using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. This technique has been used for the study of the proteomes of Candida species in the context of virulence, drug response, and resistance. We describe here the application of this approach to the study of azole antifungal resistance in C. albicans. PMID- 15888936 TI - Techniques for antifungal susceptibility testing of Candida albicans biofilms. AB - Candida albicans is capable of forming biofilms on a variety of inert and biological surfaces. Cells in biofilms display phenotypic properties that are radically different from their free-floating planktonic counterparts, including their recalcitrance to antimicrobial agents. Consequently, Candida biofilm associated infections are difficult to treat and to fully eradicate with standard antifungal therapy. Here, we describe a simple, fast, inexpensive and highly reproducible microtiter plate-based assay for the antifungal susceptibility testing of C. albicans biofilms. Because of its simplicity, compatibility with widely available 96-well microplate platform, high throughput, and automation potential, this assay represents an important tool towards the standardization of in vitro antifungal susceptibility testing of fungal biofilms. PMID- 15888937 TI - Natural products and antifungal drug discovery. AB - The need for new antifungal agents continues, fueled by opportunistic infections in immune-compromised patients and by the development of resistance to existing agents. Natural products offer a virtually unlimited source of unique molecules and not only serve as a reservoir for new potential drugs and drug prototypes, but also for probes of fungal biology. In this chapter, whole-cell screening methods targeted for natural products are illustrated, including general microplate-based screening, bioautography, and mode of action studies, including the use of genetically altered fungal strains now available commercially. PMID- 15888938 TI - Use of an animal model of disseminated candidiasis in the evaluation of antifungal therapy. AB - Animal models have been helpful in assessing a drug's potential application to treatment of humans. These controlled experiments allow description of the impact of a wide range of important treatment variables, including drug dose or concentration, dosing interval, pathogen, and host immune state. Animal models of mycoses are designed to address particular therapeutic questions. Some models are designed to screen multiple compounds in a rapid manner. Other models are selected to more closely mimic clinical infections. The following chapter will be limited to the description of a murine model of disseminated candidiasis in which in vitro results, pharmacokinetic properties, and a microbiological in vivo end point is used in the assessment of a wide variety of antifungal compounds. PMID- 15888939 TI - Murine model of invasive aspergillosis. AB - The growing importance of infectious caused by Aspergillus species during the last decade has created a need for practical and reproducible animal models of invasive aspergillosis suitable for studying fungal virulence, infection pathogenesis, diagnostic markers, and testing of antifungal therapy. Murine models remain the most commonly used models for studying aspergillosis because of their ease of manipulation and the large number of reagents available for studying disease-host responses. This chapter provides describes a murine model of invasive aspergillosis suitable for basic and translational studies of invasive pulmonary aspergillosis and highlights experimental variables that affect the course and reproducibility of infection. PMID- 15888940 TI - Antifungal combinations. AB - The increase in fungal infections and the change in fungal epidemiology is caused by the extensive use of antifungal agents to treat fungal infections that are being diagnosed in severly immunocompromised hosts. In addition, opportunistic fungal infections resistant to antifungal drugs have become increasingly common, and the armamentarium for treatment remains limited. A possible approach to overcoming these problems is to combine antifungal drugs, especially if the mechanisms of action are different. The in vitro test is the first step to evaluate possible antifungal combinations. In this chapter, the three most frequently used metholodologies are described: checkerboard, E-test, and time kill curves. The description of each technique and intrepretaion of the results are addressed in detail. PMID- 15888941 TI - Postantifungal effect methods. AB - Postantifungal effect (PAFE) is the evaluation of antifungal activity after the suppression of fungal growth when the drug is removed from the fungal suspension. In vitro, this effect might simulate the in vivo situation when the concentration of the drug falls to less than the minimum inhibitory concentration values and could be another tool, together with the classic in vitro susceptibility tests, to optimize the interaction of drugs-fungi. In this chapter, two model methods to evaluate the PAFE of yeasts and filamentous fungi are described in which practical advices and tricks are given to help the worker to develop the techniques. The procedures outlined include preparation of stock solutions of the drugs, concentration medium, exposure time colony count determination, and interpretation of the results to quantify the PAFE. PMID- 15888942 TI - Evaluation of immunotherapy in invasive candidiasis: antifungal activity and cytokine expression assays. AB - Candida albicans and non-albicans Candida spp. can cause serious infections in hospitalized and immunocompromised patients. A large number of antifungal agents and immunomodulators have been developed and may interact with both polymorphonuclear and mononuclear phagocytes against blastoconidia and pseudohyphae of Candida spp. Blastoconidia are predominantly destroyed by phagocytosis, whereas pseudohyphal killing is initiated after the attachment of phagocytes to the cell wall of the organism and subsequent extracellular release of oxidative and nonoxidative metabolites. An array of methods to evaluate phagocytosis of blastoconidia, oxidative burst, expression of cytokines and chemokines in response to Candida spp., as well as candidacidal activity are described in this chapter. PMID- 15888943 TI - Interaction of Histoplasma capsulatum with human macrophages, dendritic cells, and neutrophils. AB - Histoplasma capsulatum (Hc) is a dimorphic fungal pathogen indigenous to the Ohio and Mississippi River Valleys in the United States. Infection is initiated by inhalation of microconidia or small mycelial fragments into the terminal bronchioles of the lung. The conidia are taken up by alveolar macrophages (Mphi), in which they convert to the pathogenic yeast phase. The yeasts replicate in the alveolar Mphi and other Mphi recruited to the lung as part of the inflammatory response. Thus, the yeasts are able to disseminate from the lung to other organs, such as the liver and spleen. As a facultative intracellular parasite, the interaction of Hc yeasts with Mphi is a critical component of the host response to infection. In addition, Hc yeasts have critical interactions with inflammatory neutrophils, and with dendritic cells (DCs) in the lung and other organs. Indeed, recent new evidence suggests that DCs may be the key antigen-presenting cells that initiate cell-mediated immunity. Thus, the methods described in this chapter cover quantitation of the binding, ingestion, and intracellular replication of Hc yeasts in human Mphi, DCs, and neutrophils. PMID- 15888944 TI - Immunology of infection caused by Cryptococcus neoformans. AB - Cryptococcus neoformans is an opportunistic fungal pathogen that may lead to life threatening meningoencephalitis and pulmonary infections in immunosuppressed hosts. The lack of an effective fungicidal regimen and the development of antifungal resistant strains suggest that continued investigation is necessary to devise immunotherapeutic strategies and/or drug targets to combat C. neoformans infections. Studies to date involve investigating the host-pathogen interaction of cryptococcal infections through the genetic manipulation of the yeast, as well as the characterization of the host immune response. Macrophage phagocytosis and killing assays have proven to be invaluable tools in evaluating the putative effects of the genetic manipulation of C. neoformans strains on the virulence composite of the yeast. In addition, the assay is used to assess the efficacy of various immunotherapeutic agents (i.e., antibodies and cytokines) to enhance this cell-based antifungal activity. The purpose of this chapter is to provide a brief overview on host immunity to C. neoformans infection and, in addition, describe a protocol for performing macrophage phagocytosis and killing assays with C. neoformans and its mutants. PMID- 15888945 TI - JCO survey of orthodontic staff job satisfaction. PMID- 15888947 TI - Transforce lingual appliances for arch development. PMID- 15888948 TI - Uprighting a horizontally impacted mandibular second molar. PMID- 15888949 TI - The eight components of a balanced smile. PMID- 15888950 TI - Temporary anchorage devices: a status report. PMID- 15888951 TI - JCO interviews. Bjorn U. Zachrisson, DDS, MSD, PhD, on current trends in adult treatment, part 1. PMID- 15888952 TI - Adult treatment in the 21st century. PMID- 15888953 TI - Combined orthodontic, orthognathic, and plastic surgical treatment of an adult Class II malocclusion. PMID- 15888954 TI - Adult treatment financing: an interview with Martin (Bud) Schulman. Interview by Ron Redmond. PMID- 15888955 TI - The readers' corner. What percentage of your active patients are adults? PMID- 15888956 TI - Surgical-orthodontic treatment of two skeletal Class III patients with Invisalign and fixed appliances. PMID- 15888957 TI - The Invisalign office. PMID- 15888958 TI - Effect of L-NAME on the synthesis of plasma fibrinogen in mice. AB - The effect of nitric oxide (NO) on plasma fibrinogen was investigated by treating mice with oral N(G)-nitro-L-arginine-methyl ester (L-NAME), a non-selective NO synthase (NOS) inhibitor. Treatment with L-NAME significantly increased the concentration of plasma fibrinogen and hepatic expression of the alpha- and beta chains of fibrinogen, whereas plasma NO(2)(-) level and hepatic expression of endothelial NOS (eNOS) mRNA were significantly decreased. These results suggest that L-NAME treatment can increase plasma fibrinogen levels via an increase in expression of fibrinogen mRNA in the liver, and NO may be involved in plasma fibrinogen increase. PMID- 15888959 TI - Differences in alpha 1-adrenoceptor subtype-mediated vasoconstriction by tyramine and nerve stimulation in canine splenic artery. AB - This study was designed to clarify the alpha(1)-adrenoceptor subtypes mediating the vasoconstrictor response to tyramine in isolated and perfused canine splenic artery. It was shown that tyramine potentiated the nerve stimulation-induced second peaked vasoconstriction that was readily suppressed by prazosin treatment. A bolus injection of tyramine (0.01-0.3 micromol) caused a vasoconstriction in a dose-related manner. The tyramine-induced vasoconstriction was inhibited by WB 4101 (10 and 100 nM), an alpha(1A)-and alpha(1D)-adrenoceptor antagonist, in a concentration-related manner. Neither BMY 7378 (100 nM), a selective alpha(1D) adrenoceptor antagonist, nor chloroethylclonidine (60 microM), an alpha(1B)- and alpha(1D)-adrenoceptor antagonist, affected the tyramine-induced response. The results indicate that the noradrenaline released by tyramine may diffuse to the extrajunctional cleft, and thus it activates the extrajunctional alpha(1A) adrenoceptors, because nerve stimulation-evoked second peaked vasoconstrictions were markedly inhibited by chloroethylclonidine but not by WB 4101. PMID- 15888960 TI - The cannabinoid receptor agonist WIN 55212-2 inhibits neurogenic inflammations in airway tissues. AB - We examined the effects of a cannabinoid receptor agonist, (R)-(+)-[2,3-dihydro-5 methyl-3-[(4-merpholino)methyl]pyrrolo-[1,2,3-de]-1,4-benzoxazin-6-yl](1 naphthyl)methanone (WIN 55212-2), on various respiratory reactions induced by the activation of capsaicin-sensitive afferent sensory nerves (C-fibers). WIN 55212-2 significantly inhibited capsaicin-induced guinea pig bronchoconstriction, but not the neurokinin A-induced reaction. Intravenous injection of WIN 55212-2 also blocked cigarette smoke-induced rat tracheal plasma extravasation. However, substance P-induced rat tracheal plasma extravasation was not affected by the administration of WIN 55212-2. A cannabinoid CB(2) receptor antagonist, {N-[(1S) endo-1,3,3-trimethylbicyclo[2.2.1] heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4 methylbenzyl)pyrazole-3-carboxamide} (SR 144528) reduced the inhibitory effects of WIN 55212-2, but not a cannabinoid CB(1) antagonist, [N-(piperidin-1-yl)-5-(4 chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3 carboxamidehydrochloride] (SR 141716A). A Maxi-K(+) channel opener, 1-(2'-hydroxy 5'-trifluoromethylphenyl)-5-trifluoromethyl-2(3H)benzimidazolone (NS 1619), specifically inhibited capsaicin-induced guinea pig bronchoconstriction and cigarette smoke-induced rat tracheal plasma extravasation. These findings suggest that WIN 55212-2 inhibits the activation of C-fibers via cannabinoid CB(2) receptors and Maxi-K(+) channels and reduces airway neurogenic inflammatory reactions in vivo. PMID- 15888961 TI - Detailed pharmacological characterization of 5-HT1A-receptor-mediated [35S]GTP gamma S binding in rat hippocampal membranes. AB - 5-HT-stimulated [(35)S]GTPgammaS binding to rat hippocampal membranes was pharmacologically characterized. Signal/noise ratio or percent increase over basal was optimized with 100 microM GDP, 2-10 mM MgCl(2), and 150-200 mM NaCl. However, we preferred the standard condition (20 microM GDP, 5 mM MgCl(2), and 100 mM NaCl: Condition I) to the alternative one (100 microM GDP, 5 mM MgCl(2), and 150 mM NaCl: Condition II) because 1) absolute values of basal and 5-HT sensitive bindings decreased with higher concentrations of GDP and NaCl; 2) EC(50) values determined under Condition II were 2 - 6 fold higher than those under Condition I; 3) some partial agonists had less intrinsic activities in the presence of higher concentrations of GDP; and 4) Inhibitory effects of WAY100635 were complete under Condition I, while incomplete under Condition II. Pharmacological profile of concentration-dependent stimulation by a series of 5 HT ligands and concentration-dependent inhibition of 5-HT-stimulated binding by several 5-HT-receptor antagonists clearly indicated that this response under Condition I was mediated solely through 5-HT(1A) receptors. Although caution should be paid especially to the apparent intrinsic activities susceptible to the assay conditions, this method appears useful to investigate functional coupling between 5-HT(1A) receptors and their coupled G proteins in native hippocampal membranes. PMID- 15888962 TI - Corymine potentiates NMDA-induced currents in Xenopus oocytes expressing NR1a/NR2B glutamate receptors. AB - Previous studies demonstrated that corymine, an indole alkaloid isolated from the leaves of Hunter zeylanica, dose-dependently inhibited strychnine-sensitive glycine-induced currents. However, it is unclear whether this alkaloid can modulate the function of the N-methyl-D-aspartate (NMDA) receptor on which glycine acts as a co-agonist via strychnine-insensitive glycine binding sites. This study aimed to evaluate the effects of corymine on NR1a/NR2B NMDA receptors expressed in Xenopus oocytes using the two-electrode voltage clamp technique. Corymine significantly potentitated the NMDA-induced currents recorded from oocytes on days 3 and 4 after cRNA injection but it showed no effect when the current was recorded on days 5 and 6. The potentiating effect of corymine on NMDA induced currents was induced in the presence of a low concentration of glycine (< or =0.1 microM). Spermine significantly enhanced the potentiating effect of corymine observed in the oocytes on days 3 and 4, while the NMDA-receptor antagonist 2-amino-5-phosphonopentanone (AP5) and the NMDA-channel blocker 5 methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801) reversed the effect of corymine. On the other hand, the nonspecific chloride channel blocker 4,4-di-isothiocyano stilbene-2,2-disulfonoc acid (DIDS) had no effect on the corymine potentiation of NMDA currents. There was no good correlation between corymine- and spermine-induced potentiation of the NMDA-current response in Xenopus oocytes. These results suggest that corymine potentiates the NMDA-induced currents by interacting with a site different from the spermine binding site. PMID- 15888963 TI - Chronic heavy ethanol consumption is associated with decreased platelet aggregation in rats. AB - Although moderate alcohol consumption seems to be protective against atherosclerosis, coronary artery disease rate increases with its higher doses. Platelet aggregation is an important process which contributes to the atherosclerosis. The aim of this study was to determine whether heavy ethanol consumption stimulates or inhibits platelet aggregation. Fourteen adult male Wistar rats were used. Ethanol (7.2%, v/v) in a modified liquid diet was given to eight rats for 21 days, which mimicked characteristics similar to human chronic alcoholism. Six rats constituted the control group. Adenosine diphophate (ADP) and collagen-induced platelet aggregation was measured in whole blood. We found reduced ADP-induced mean maximal aggregation in the alcoholic rat group compared to the control group at dose of 5 microM (p < 0.005). We also found decreased platelet aggregation responses to collagen in the alcoholic group (p < 0.006 for 2 microg/ml collagen, and p < 0.05 for 5 microg/ml collagen). In conclusion, chronic heavy ethanol consumption results in the decreased platelet aggregation in a rat model of alcoholism. Therefore, increased mortality from coronary artery disease in chronic alcoholism may be explained by other factors such as dietary imbalances and coexisting conditions, which include hypertension and depression. PMID- 15888964 TI - Assessment of acute phase proteins in acute ischemic stroke. AB - Acute phase proteins (APPs) have been implicated to play important roles during both acute and chronic inflammatory processes in different diseases including ischemic stroke. Though there are several studies showing the importance of APPs as inflammation markers in acute ischemic stroke (AIS), the time course of these proteins during acute phase of AIS is not well known. Thus, the aim of this study was to show the changes in plasma levels of six APPs (i.e., haptoglobin [Hp], ceruloplasmin [Cp], high-sensitive C-reactive protein [h-CRP], fibrinogen, complement 3 [C3] and complement 4 [C4]) during the first 10 days after acute stroke. The study group consisted of 34 female and 19 male patients (n = 53; mean age 65 +/- 12 years), who had first acute ischemic stroke (AIS). An age-matched control group (n = 53; 32 female and 21 male subjects, mean age 62 +/- 6 years) was also included. To evaluate the plasma levels of six APPs, the blood samples of patients with AIS were withdrawn on admission (day 1), and after 3, 5 and 10 days, whereas only one measurement was performed in the control group. In addition, several cerebrovascular risk factors were determined. The peak levels of APPs were higher in the AIS group than the control group (p < 0.0001). In serial measurements, the levels of h-CRP, Hp, C3 and C4 showed alterations during 10 days after AIS (p < 0.0001, p < 0.05, p < 0.0001, p < 0.0001, respectively). The alterations in levels of fibrinogen and Cp were not statistically significant (p > 0.05). After stroke, h-CRP, C3 and fibrinogen reached their highest values on the third day, Cp and C4 on the fifth day, and Hp on the tenth day. The plasma levels of h-CRP correlated positively with other five APPs studied (p < 0.05). These findings support the importance of inflammation processes after stroke. We suggest that the differences in levels of APPs could be used in predicting the outcome of stroke patients. PMID- 15888965 TI - Bowel habits before and during menses in Japanese women of climacteric age: a population based study. AB - A good deal of data are available on the bowel habits of pre-climacteric females during menstruation. Few studies have examined the same subject in females of climacteric age but who are still menstruating, so the present study was undertaken to examine the bowel habits in menstruating females in this age group. Subjects (n = 246) were residents of a city in northern Japan, aged from 45 to 55 years old and who were still menstruating. Their every-day state of defecation and fecal characteristics were assessed with regard to four parameters: bowel movement frequency, defecation state, fecal appearance and fecal consistency. Based on a perfect match to these four criteria, the subjects were assigned to two groups, the normal group and the constipation group, and changes were assessed by enquiring how their bowel habits differed immediately before and during menses compared with their usual state. Immediately before menstruation, in the constipation group feces became harder and looser in 22.1% and 13.7%, respectively, compared with 8.7% and 9.5% in the normal group. On the other hand, during menstruation in the constipation group, feces became harder and looser in 11.6% and 16.8%, respectively, compared with 5.7% and 8.9% in the normal group. In other words, the constipation group showed a greater change immediately before and during menstruation compared with the normal group, though the differences were not significant. Our data thus suggest that the changes in the bowel habits of women of climacteric age at menstruation are greater for those suffering from constipation than those who are not. PMID- 15888966 TI - No association of the CAG repeat length in exon 1 of the androgen receptor gene with idiopathic infertility in Turkish men: implications and literature review. AB - While the correlation between the CAG repeat length of the androgen receptor (AR) gene and idiopathic male infertility is still unclear, ethnic background of the population studied may play an important role in this association. The objective of this study was to determine whether changes in the CAG repeat length are associated with spermatogenic defects in Turkish infertile men. Reproductive hormone concentrations and the CAG repeat length in exon 1 of the AR gene in 47 idiopathic infertile men and 32 fertile controls were analyzed. The mean serum luteinising hormone (LH) and follicle stimulating hormone (FSH) levels were significantly higher in the infertile group than those of the control group (p < 0.0001 for both comparisons), whereas the mean serum testosterone level in the infertile group did not differ significantly from that in the control group (p = 0.16). The mean CAG repeat length of the AR gene in the infertile group did not differ significantly from that in the control group (22.28 +/- 0.37 vs 22.41 +/- 0.54, respectively; p = 0.84). In addition, the frequency of having a CAG repeat number (> or = 24) was also comparable between the infertile patients and fertile controls (31.9% vs 40.6%, respectively; p = 0.21). In conclusion, reproductive hormones with elevated LH and FSH, and normal or low testosterone levels were suggestive of partial impairment of testicular function. However, no statistically significant relationship between the length of the CAG repeat and idiopathic impaired sperm production was observed in the Turkish population studied. These results support the findings of previously published European studies, but are contrary to the findings from Caucasian and North American population studies. Thus, ethnicity and genetic backgrounds seem to be important in this association, and studies from a variety of different ethnic and genetic backgrounds using comparable patient subgroups are valuable to further evaluate this association. PMID- 15888967 TI - Social adaptation of children with congenital fecal dysfunction: from the viewpoint of the mother-child relationship. AB - Excretory dysfunction associated with congenital anal anomalies (a generic term that includes anal atresia and Hirschsprung's disease) is presumed to greatly affect the psychology of the affected children. In this study, we conducted a survey on the psychological status of children with excretory dysfunction, and investigated the relationship between the affected children and their families in addition to their social life. Four children with fecal dysfunction due to a congenital anal anomaly and their mothers were interviewed. The results of psychological tests in the children and mothers along with our findings in the interviews with the children and mothers, were included in the data analysis. We found that regardless of the degree of excretory dysfunction, the child's state of mind was influenced by whether the mother exhibited warmth or criticism towards her child and whether there was a support system for the mother and child. We suggest that psychiatric consultation is necessary for these children. PMID- 15888968 TI - Aural myiasis in children and literature review. AB - Myiasis is a disease caused by fly larvae. The term "myiasis" is derived from the Greek word "myia" meaning fly. Aural myiasis is a rare clinical state and occurs frequently in children. In this article, six children with aural myiasis, caused by the fly larvae, are reported because of unusual presentation. All of the children with aural myiasis were associated with chronic otitis media. In the treatment, a combination of suctioning and alligator forceps was used to remove maggots under the light microscopic field. Additionally, antibiotics were used in all children. Thus, aural myiasis is successfully treated by direct extraction of larvae and application of preventative methods. PMID- 15888969 TI - Beneficial effects of N-acetylcysteine on acetic acid-induced colitis in rats. AB - Ulcerative colitis is a chronic recurrent inflammatory bowel disease in which oxidative stress has been implicated. The aim of the present study was to evaluate possible protective effects of N-acetylcysteine against acetic acid induced colitis in a rat model. Rats were administered intrarectal saline (control group) or acetic acid (colitis model group). Rats with acetic acid induced colitis were treated by intraperitoneal or intrarectal administration of N-acetylcysteine (500 mg/kg) (treated group). Another series of rats were pre treated by intraperitoneal or intrarectal administration of N-acetylcysteine, then administered intrarectal acetic acid (pre-treated group). The degree of tissue injuries was assessed by macroscopical and histopathological scores of the colonic mucosa. Malondialdehyde, myeloperoxidase, reduced glutathione, superoxide dismutase, and catalase levels were measured in tissue extracts of the dissected colon. Administration of N-acetylcysteine intraperitoneally or intrarectally ameliorated macroscopic score alterations produced by acetic acid in treated groups. In addition, microscopical improvement was observed in all N acetylcysteine-treated rats compared to untreated animals with colitis. In the colonic tissues of the acetic acid-induced colitis, myeloperoxidase activity and malondialdehyde levels were elevated, while the reduced glutathione levels and superoxide dismutase and catalase activities were decreased. However, intraperitoneal or intrarectal treatment with N-acetylcysteine reversed these parameters, compared to the untreated colitis group. Notably, intrarectal administration of N-acetylcysteine elevated the reduced glutathione levels more markedly compared to the other treatment groups. Superoxide dismutase levels were increased in intraperitoneally or intrarectally N-acetylcysteine-treated groups significantly compared to the control, colitis and pre-treated groups. But there was no significant increase in catalase activity. In conclusion, N-acetylcysteine could be beneficial as a complementary agent in treatment of ulcerative colitis. PMID- 15888970 TI - Suppressive effects of red wine polyphenols on voltage-gated ion channels in dorsal root ganglionic neuronal cells. AB - Polyphenols are ubiquitous plant metabolites with multiple pharmacological properties. Using whole-cell patch-clamp current recording techniques, we studied the effects of polypnenols extracted from red wine (purity > 90% from Cabernet Sauvignon grape wine) on the activities of voltage-operated Na+-, K+-, and Ca2+ channel currents in mouse dorsal root ganglionic neuronal cells. The polyphenols suppressed all of the channel activities with half-effective concentrations of about 2.5, 4.0, and 0.8-1.5 micro g/ml, respectively. In contrast, they showed no noticeable effects on the ion channels in other types of cells, including large conductance K+-channels in mouse lacrimal acinar cells. Thus, the polyphenols suppress firings of the action potential in the neuronal cells and could show a sedative effect on the excitation. We expect that red wine can be used as a remedy for excessive sensory stimuli. PMID- 15888971 TI - Open-ended questions: are they really beneficial for gathering medical information from patients? AB - Open-ended questions, which allow patients to discuss their concerns freely, are widely considered an efficient method gathering medical information from patients during a medical interview. However, few studies have examined the relationship between the use of open-ended questions and the amount of information obtained from patients during the medical interview. This study examines this relationship using a relatively large sample size under more standardized conditions than in previous studies. The Japanese Group for Research on the Medical Interview undertook this research in 2002-2003. A total of 1,527 medical students conducted medical interviews with standardized patients, and 1,220 met the inclusion criteria for this study. The interview was limited to five minutes. Evaluators (medical school faculty physicians) evaluated the use of open-ended questions during the medical interview. The reliability of the evaluation sheet was also examined. The amount of information obtained was measured through the medical interview evaluation sheet. The use of open-ended questions was positively related to the amount of information elicited from the patients (F = 41.0, p < 0.0001). This study provides data to support the hypothesis regarding the favorable relationship between the use of the open-ended questions and the amount of information from the patients. PMID- 15888972 TI - Protective effects of vitamin C, alone or in combination with vitamin A, on endotoxin-induced oxidative renal tissue damage in rats. AB - This study was designed to investigate the protective effects of vitamin C and vitamin A on oxidative renal tissue damage. Male Wistar rats were given an intraperitoneal injection of 0.5 ml saline (control) or 0.5 ml solution of lipopolysaccharide (10 mg/kg), which caused endotoxemia. Immediately (within 5 min) after the endotoxin injection, the endotoxemic rats were untreated or treated with intraperitoneal injection of vitamin A (195 mg/kg bw), vitamin C (500 mg/kg bw) or their combination. After 24 hours, tissue and blood samples were obtained for histopathological and biochemical investigation. Endotoxin injection caused renal tissue damage and increased erythrocyte and tissue malondialdehyde (MDA) and serum nitric oxide (NO), urea and creatinine concentrations, but decreased the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities compared to the parameters of control animals. Treatment with vitamin C or with vitamins C and A significantly decreased the MDA levels and serum NO, urea and creatinine levels, recovered the antioxidant enzyme activities (SOD, GSH-Px and CAT), and prevented the renal tissue damage in endotoxemic rats. In contrast, vitamin A alone did not change the altered parameters except for creatinine levels. Notably, the better effects were observed when vitamins A and C given together. It is concluded that vitamin C treatment, alone or its combination with vitamin A, may be beneficial in preventing endotoxin-induced oxidative renal tissue damage and shows potential for clinical use. PMID- 15888973 TI - Nuclear factor-kappaB activation in diabetic rat kidney: evidence for involvement of P-selectin in diabetic nephropathy. AB - Activation of a transcription factor, nuclear factor-kappaB (NF-kappaB), is a key step in the pathogenesis of diabetic nephropathy. In this study, we investigated the role of P-selectin, a platelet-derived adhesion molecule, in diabetic nephropathy by examining the activation status of NF-kappaB in the renal cortex of streptozotocin (STZ)-treated rats. The STZ treatment induced pathogenetic parameters such as increased creatinine clearance, increased blood glucose and massive albuminuria in a time-dependent manner. Electrophoretic mobility shift assays (EMSAs) with a specific probe, representing the P-selectin gene promoter, revealed the activation status of NF-kappaB in the STZ-treated rats, as judged by the time-dependent increase in the formation of the specific protein-DNA complexes. This increase was associated with the increased pathogenetic parameters. Supershift assays with specific antibodies revealed that p50, but not p52, p65, Rel B, or c-Rel, may be involved in the activation of NF-kappaB, though the component primarily responsible for the increase could not be determined. Western blot analysis confirmed an increase in P-selectin in STZ-treated rats. Notably, treatment with ammonium pyrrolidinedithiocarbamate, an antioxidant and inhibitor of NF-kappaB, inhibited the activation of NF-kappaB in STZ-treated rats and decreased P-selectin in the renal cortical tissue. Our results indicate that expression of the P-selectin gene is induced through the activation of NF-kappaB and that P-selectin may be involved in the pathogenesis of diabetic nephropathy. PMID- 15888974 TI - A patient with clinical features of acute hepatitis E viral infection and autoimmune hepatitis. AB - Hepatitis E virus (HEV) is one of the major causative agents of acute hepatitis in many developing countries. Recent intensive examination has revealed the existence of non-imported cases in industrialized countries. The patient was a 25 year-old Japanese female with acute hepatitis. Laboratory test demonstrated positive anti-nuclear antibody (ANA), anti-smooth muscle antibody (ASMA) and high level of serum immunoglobulin G (IgG). The patient was negative for serum markers of hepatitis A, B or C virus infection. She demonstrated a clinical course similar to severe autoimmune hepatitis, including response to prednisolone therapy. After a few years, with the availability of tests for the serum antibodies to HEV, we examined the frozen stocked sera of the patient and found her exact diagnosis was acute hepatitis E. Although we could not detect HEV-RNA, which is positive only in limited period of acute phase, serum IgA and IgG antibodies to HEV were positive and the titer of IgA and IgG antibodies were declined with the time course. In conclusion, we must take into consideration of HEV infection for the diagnosis of acute cryptogenic hepatitis including autoimmune hepatitis. Further studies are feasible to understand the pathogenesis of liver injuries induced by HEV infections. PMID- 15888975 TI - Pleomorphic carcinoma of the lung associated with loss of heterozygosity of p53 gene. AB - We report a case with pleomorphic carcinoma of the lung in a 70-year-old man. Pleomorphic carcinoma is characterized by a heterogenous composition that includes epithelial and mesechymal malignancies. In the present case, the tumor was composed of a mixture of unequivocal squamous cell carcinoma and spindle cell components resembling sarcomatous overgrowth. The spindle component did not include a heterologous mesenchymal element characterized by overt differentiation for bone, cartilage, neuron or muscle tissue. To evaluate a state of differentiation of the spindle cell component, we immunohistochemically examined expression of the antigens including vimentin, cytokeratin, sarcomeric actin, alpha-smooth muscle actin, S-100 protein, CD34, Factor VIII, and CD68. The results showed sole expression of vimentin in the spindle cell component, suggesting an immature state of the mesenchymal lineage. Furthermore, the spindle cell component of this case was genetically characterized by loss of heterozygosity (LOH) at a codon 234 of exon 7 of the p53 gene. This mutation causes an amino-acid replacement (Tyr to Cys), which was previously proven to attenuate p53 function. The present case may suggest a relation between somatic alteration of the p53 gene and histogenesis of pleomorphic carcinoma. PMID- 15888976 TI - [Pulmonary hypertension and HIV: implementation of a Regional Registry]. AB - In the era of new, potent antiretroviral therapy, much more attention is being given to non-infectious complications of HIV diseases, such as cardiomyopathy, pericardial effusion and pulmonary hypertension (PH). PH diagnosis is based on a mean pulmonary artery pressure of more than 25 mmHg at rest, or more than 30 mmHg with exercise. The incidence of PH is about 0.1% per year among HIV-positive patients, while in the general population it is 1 to 2 cases per million people. The histopathology of HIV-associated PH (HAPH) is similar to that of idiopathic PH, although its pathogenesis is still unclear. In patients with HAPH secondary causes of PH must be ruled out, such as intravenous drug abuse, valvulopathy, congenital heart disease and previous tricuspid endocarditis. The treatment of HAPH is not substantially different from that of idiopathic PH and is essentially based on the use of vasodilators. The Regional Authority of Lazio (Italy) has instituted a Registry for PH in HIV-positive patients; its aims are to evaluate the real incidence and prevalence of primitive and secondary PH among patients with HIV infection, and optimise the management of patients with suspected PH through the definition of a diagnostic algorithm. PMID- 15888977 TI - [An attempt to improve classification of HCV-correlated chronic hepatitis]. AB - To verify the clinical efficacy of the Desmet classification of chronic hepatitis C we reviewed 801 liver biopsies from patients with HCV-chronic hepatitis (CH). The diagnosis of chronic hepatitis was assessed according to the Desmet classification based on the Knodell Histological Activity Index (HAI) (minimal CH=score 1-3; mild CH= 4-8; moderate CH= 9-12; severe CH= 13-18). Liver fibrosis was assessed according to the Scheuer scoring system. One hundred forty-eight patients had cirrhosis and 653 CH. Of these 653, according to the Desmet classification 145 patients showed minimal, 424 mild, 73 moderate and 11 severe chronic hepatitis. Since the classification underestimated the moderate and severe forms of HCV-related chronic hepatitis, we evaluated the possibility of improving the Desmet classification of chronic hepatitis C using our classification: minimal CH= score 1-3; mild CH= 4-6; moderate CH= 7-8; severe CH= 9-18. According to our classification 145 showed minimal CH, 363 mild CH, 61 moderate CH and 84 severe CH. All the 61 patients who crossed over from mild CH under the Desmet to moderate CH under our classification showed a periportal inflammation of grade 3, and all the 73 patients but 8 who crossed over from moderate to severe showed a grade of periportal inflammation higher than 3. The Desmet classification of HCV-related chronic hepatitis underestimated the severe forms of HCV-CH, while our classification seems to be suitable also for chronic hepatitis C. PMID- 15888978 TI - [Alterations of bone metabolism in patients with chronic C virus hepatitis]. AB - Patients affected with chronic hepatitis are prone to alterations in bone metabolism, osteoporosis and osteopoenia being the most common manifestations. Bone mineral densitometry is the method of choice for assessing bone mass; nevertheless, this is a static parameter whereas biochemical markers of bone remodelling reveal the dynamics of bone resorption and formation. With this study we used bone mineral densitometry and biochemical markers to evaluate bone metabolism in a group of male patients with chronic C virus hepatitis and in a group of healthy males. In the hepatitis group 56% of the patients proved osteopoenic or osteoporotic and bone depletion increased as the histological score of the disease increased. Crosslaps are a parameter of osteoclastic activity: their measurement showed alterations in all the age groups of the hepatitis patients studied, which goes to show that there is intense bone remodelling in these individuals due mainly to osteoclastic resorption. Hepatitis C is a risk factor for bone depletion: we believe that when this type of hepatitis is diagnosed it is useful to assess bone metabolism with bone mineral densitometry and with the crosslaps assay. PMID- 15888979 TI - Clinical and laboratory features of acute rheumatic fever: a 18-year experience. AB - We present the retrospective analysis of clinical manifestations and laboratory findings observed in 30 patients (M/F 13/17; age range 9-66 yrs) affected by acute rheumatic fever observed within the Infectious Disease Department along a period of 18 years (1986-2004). Diagnosis of carditis was stated on clinical and echocardiographical bases and occurred in 50% of patients. Such patients presented mild to moderate heart disease (30%) and severe carditis (20%). Therefore, our data stand to confirm that rheumatic cardiac disease could determine permanent and/or severe heart damage. All patients were observed during a 48-month period of follow-up without exitus. PMID- 15888980 TI - [Evolution in the hospitalization for infectious diseases among non-EU patients in Emilia Romagna]. AB - In the Emilia Romagna (ER) area, between 1996 and 2000, a progressive increment in hospitalization for TBC, malaria, AIDS and hepatitis in non-EU patients was observed. This study aims to determine whether this trend was confirmed in 2001 and in which cities the increase was most significant. The Hospital Discharge Cards (HDC) registered in ER for non-EU patients in the relevant period were examined. In 2001, of 20,980 hospitalization cases of non-EU patients, 394 (1.87%) were attributed to infectious diseases, amounting to an increase of 1.77% over 2000. Of the 394 patients 250 (63.45%) were male and 144 (36.55%) female. The most represented age group was 20-39 yrs. Male patients more frequently come from Morocco (54), Senegal (45), Brazil (43), females from Nigeria (36), Morocco (26) and Ghana (14). The towns and cities where hospitalization occurred were, in decreasing order: Modena (24.6%), Bologna (19.3%), Reggio Emilia (12.9%), Ravenna (10.4%), Rimini (8.6%), Parma (8.3%), Piacenza (7.3%), Forli (4.8%), Ferrara and Cesena (both 1.8%). The Hospital Departments primarily involved were: Infectious Diseases with 213 hospitalizations (54%), Pneumology 69 (17.5%), Medicine 44 (11.1%), and Paediatrics 39 (9.9 %). Hospitalization causes were, in order of frequency: TBC with 137 cases (34.8%), malaria 75 cases (19%), AIDS 72 cases (18.3%), viral hepatitis 56 cases (14.2%), septicaemia 22 cases (5.6%) and Salmonella spp. infections 18 cases (4.5%). PMID- 15888981 TI - A case of right-side infective endocarditis with ventricular septal defect. AB - A 28-year-old woman previously known to have a ventricular septal defect presented with fever, headache, abdominal pain and nausea. Positive blood culture of methicillin-sensitive Staphylococcus aureus and the detection of vegetation attached to the right ventricular wall near the ostium of the ventricular septal defect confirmed diagnosis of infective endocarditis. After four weeks' treatment with proper antibiotics the patient recovered. PMID- 15888982 TI - HCV infection and pericarditis: an extrahepatic manifestation? AB - The authors describe a clinical case in which they found the unusual combination of acute hepatitis caused by HCV and pericarditis in a young person, resulting in complete recovery from the pericarditis but in a deterioration of the chronic HCV. A close examination of the literature on this subject revealed that, although no similar case was recorded, an aetiological relationship between the hepatitis C virus and pericarditis cannot be excluded since an HCV infection often gives rise to extra-hepatic cardiac problems. PMID- 15888983 TI - Acute transverse myelitis and hepatitis C virus. AB - In the last few years, substantial evidence has been provided on peripheral nervous involvement in infection by hepatitis C virus (HCV), whilst central nervous involvement is rare. Here, we report a case of acute transverse myelitis in a woman developing 4 years after documented HCV seropositivity, associated with intrathecal anti-HCV protein IgG. Isoelectrofocusing of all CSF samples before and after immunoabsorption with recombinant structural HCV proteins revealed disappearance or marked decrease of some oligoclonal IgG bands suggesting binding to HCV proteins. To our knowledge, this is the first report of acute myelitis associated with intrathecal immune response against HCV proteins. This finding suggests that in acute myelitis of unknown aetiology, testing CSF for HCV RNA and related antibodies is warranted. PMID- 15888984 TI - [Tuberculosis in the 19th and 20th centuries: epidemiology and the role of urban health programmes in tuberculosis prevention in Bologna]. AB - The author reports the scenario in which the bacteriological revolution arose and developed during the 19th century. In this period medical research improved its theoretical basis and clinical practice, and aimed at becoming an exact science. Scientific discoveries in the field of microbiology and growing social and sanitary awareness conferred a very particular specificity on tubercular disease: this pathology was defined a work-induced infectious disease, as L. Devoto stated at the beginning of the twentieth century. After introductory statements, we present statistical and epidemiological research carried out in the 19th century among populations of workers, and studies that show the relationship between the diffusion of the tuberculosis epidemic and the high concentration of people in degraded urban areas. We also present statistics on tuberculosis mortality in the Kingdom of Italy, the region of Emilia Romagna, and in Bologna. Finally, we briefly describe the urban improvement measures performed in the Bologna area after the unification of Italy (1860), which allowed contagion to be reduced, as well as both the mortality and morbidity rate in this urban area. PMID- 15888985 TI - [EPH (Epidemiology and Public Health) and the Internet]. PMID- 15888986 TI - Trends in cannabis use and its determinants among teenagers in the French speaking community of Belgium. AB - BACKGROUND: Cannabis consumption among teenagers has undergone dramatic changes in Europe since the beginning of the 1990s. A number of behaviors associated with cannabis consumption, such as tobacco smoking, excessive drinking and truancy are developing too, each in their own way. METHODS: To assess the evolution over time of the various types of cannabis consumption (both ever and weekly consumption) in relation to these determinants (age, sex, studies chosen, truancy, tobacco smoking and recurrent intoxication), we have analyzed the cross-sectional study on Health Behaviour in School-Aged Children in the French-speaking Belgian Community (12-17 years) since 1994. We used logistic models to analyze the evolution of the various types of cannabis consumption and to identify the associated factors. Finally, in order to demonstrate time trends, we tested for each type of consumption in the interactions between the significant predictive variables in each model and the survey year (1994-1998-2000). RESULTS: Rates of ever use, past 30-day use and weekly use among the ever users have been increasing from 1994 to 2002 and reached, respectively, 22.0%, 11.6%, 6.8% and 32.9%. Cannabis ever use rose more noticeably among the general education students (adjusted OR (95%CI)): 3.08 (2.66-3.57) and among the truants: 4.57 (3.39-6.14). Weekly cannabis smoking rose most especially among the truants: 1.92 (1.34-2.78). CONCLUSION: Truants should constitute a priority target for the prevention of cannabis consumption, while the phenomenon of truancy must be moreover examined in depth in order to more thoroughly identify the appropriate prevention programs organized both in and outside of the school environment. PMID- 15888987 TI - [Variant Creutzfeldt-Jakob disease in France: estimating the number of cases related to travel to the United Kingdom between 1980 and 1995]. AB - BACKGROUND: The outbreak of variant Creutzfeldt-Jakob disease (vCJD) cases rose serious concerns about secondary transmission of the disease, particularly through blood transfusion. Protective measures leading to the exclusion of potentially infectious blood donors were settled: in France, donors who had stayed more than one year in the UK were excluded. In this work, which was part of a larger study aiming to estimate the French epidemic of vCJD, the number of vCJD cases who were infected during a trip to the UK was estimated. Those estimates may notably enable the assessment of such exclusion measures. METHODS: The particular age-related structure in vCJD cases is taken into account in our simulations considering birth cohorts in the population. The total French exposure is simulated assuming the main source of infection to be dietary through consumption of mechanically recovered meat (MRM) manufactured from British bovine carcasses. Then, using a "back calculation" algorithm, all infected individuals required to produce a consistent epidemic (6 vCJD cases in 2003) was simulated. This study was exclusively focused on the part of the exposure linked to trips (beef MRM consumed in the UK while traveling) and on cases resulting from this exposure. RESULTS: The influence of exposure linked to trips to the UK was greater in the youngest cohort (6.3% of the total exposure) while it only accounted for 3.3% and 1% in the 1939-69 and in the pre-1939 birth cohorts respectively. Overall, exposure resulting from trips in the UK can be neglected with regards to the exposure linked to the consumption of MRM produced in France from British bovine carcasses. Consequently, French vCJD cases that would have been infected in the UK are very unlikely to occur (median: 0 case, IC 95%: (0 2)). Nevertheless, if such cases occur, they would probably occur in subjects born after 1969 and their onset would take place before 2010. Thus, unlike the situation in BSE-free countries, the causal relationship between travel in the UK and occurrence of vCJD cases cannot be underlined in France, as trips only account for a small part of the French exposure. CONCLUSION: Since trips in the UK slightly contribute to the overall French exposure, excluding people who travelled in the UK from blood donation would not influence the risk of secondary transmission. PMID- 15888988 TI - [Knowledge of hepatitis B and C among underprivileged people]. AB - BACKGROUND: Prevalence for hepatitis B (HBV) and C (HCV) viruses infection are particularly high among underprivileged persons (UPP) related to the use of injecting drug, residence in high endemic countries or a low rate of condom use in case of multiple sexual partners. It is important to know whether these persons are well aware of prevention measures for these infections. The aim of our study was to evaluate the knowledge of preventive measures among UPP and, at the same time, among health- and social-care workers (H/S-CWs). METHODS: The study was conducted in 68 salvation settings. The H/S-CWs filled out once 9-item self-administered questionnaires on HBV and HCV, concerning the illnesses, and their transmission, prevention, and screening. A similar questionnaire was proposed by H/S-CWs, to UPP after each interview. All answers were grouped into 4 categories: true (one right item or more), false, unknown, no answer. RESULTS: One thousand nine hundred twenty one questionnaires were collected: 312 were from H/S-CWs and 1609 from UPP. The answer rate was 100% for H/S-CWs and 92.7% among the UPP population. Knowledge about HBV and HCV was better for H-CWs compared to S-CWs. For HBV, routes of transmission was the best known field (85.2% of right answers) and use of blood sample screening was the least known field (54.2%). Vaccination was advanced by 50.6% of H/S-CWs. For HCV, routes of transmission were also the best known field (65.9%) while preventive measures were frequently unknown (22.0%). Among the UPP, blood sample screening was the best known field for HBV (34.5% of right answers) as for HCV (27.5%). Knowledge about the routes of transmission and the populations at risk was very low for HBV (28.2% and 15.5%) and for HCV (9.4% and 5.1%). The analyses of each response showed that the rates of wrong answers were much higher for H/S-CWs compared to UPP. On the other hand the rates of "unknown" answer were higher in the UPP. This finding suggests that H/S-CWs had a false perception of knowing relevant information on HBV and HCV. The high proportion of no answer among UPP confirmed their ignorance of those diseases. CONCLUSION: The UPP needs appropriate information on HBV and HCV. This information could be delivered by H/S-CWs. However, preliminary training, particularly for S-CWs, might improve their insufficient knowledge at the present time. PMID- 15888989 TI - [Cluster sampling: consequences of data analysis on drawing conclusions]. AB - BACKGROUND: Cluster sampling is commonly used since it does not require a sampling frame which lists all the individual enumeration units. However, this sampling design is often less precise than simple random sampling due to frequent homogeneity of individuals within clusters. This note illustrates that the precision of parameters such as mean, prevalence and odds ratio can be biased when the data analysis ignores the sampling design, yielding to possibly erroneous conclusions. METHODS: Data from a cluster sampling among clandestine sex workers in Senegal were used. Two analyses were performed and their results were compared. The first analysis took into account the sampling design (design based analysis) while the second did not (naive analysis). RESULTS: The range of confidence intervals in design-based analysis differed from -43% to +84% with regard to those of naive analysis, and different conclusions could be drawn. For instance, the human immunodeficiency virus (HIV) infection in clandestine sex workers was associated with condoms use and perceived risk of HIV infection in design-based analysis but not in naive analysis. CONCLUSION: The data analysis must take into account the sampling design, and this is facilitated by the availability of statistical software with survey analysis capabilities. PMID- 15888990 TI - [Time-dependent covariates in the Cox proportional hazards model. Theory and practice]. AB - In survival analysis, exposure that appears or changes during the follow-up of subjects must be taken into account as a time-dependent covariate in the Cox proportional hazards model. Two types of time-dependent covariates are defined: covariates with unique change, and covariates with multiple changes. The way of taking into account such changes in the exposure is presented in theory and illustrated from a small sample of 5 subjects enrolled in a French HIV cohort. The problems raised by missing data as well as alternative but more sophisticated solutions are also evocated. Annexes include programs for SAS software, and results from the "Log" and "Results" windows. PMID- 15888991 TI - [Factors associated with cervical cancer screening in women covered by national health insurance]. AB - BACKGROUND: The aims of our study were to examine the factors associated with cervical screening in women whose tests are supported by a national health insurance scheme, the Mutuelle Generale de l'Education Nationale. METHODS: A self administered postal questionnaire was sent to a random sample of 10,000 adults aged 20 to 60 years old living in France and insured with the Mutuelle Generale de l'Education Nationale. Response rate was 66.5% (N=6,518). RESULTS: Of the 3,741 women aged 20 to 60 years old, 88.5% have had at least one cervical smear. Mean age for the first cervical smear was 29.3 years (95% CI: 29.0-29.6). Preventive practices (mammogram, cervical smear and fecal occult-blood testing) were strongly related. The predictive factors for cervical smear during the past three years included: age between 30 and 49 years, marital status (married, separated or divorced), the socioeconomic status (unskilled workers reported cervical smears less often than women from other socioeconomic status). Consulting a gynaecologist in the past twelve months increased 9-fold the probability of having a cervical smear and 2-fold when consulting a general practitioner compared to women who consulted neither a gynecologist nor a general practitioner. CONCLUSIONS: In a population whose tests were supported by a national health scheme, the socioeconomic status was one of the determinants of Pap smear. PMID- 15888992 TI - [First results from the ANRS-VESPA survey of persons living with HIV/AIDS]. PMID- 15888993 TI - Novel risk factors for heart disease. An interview with Gerald Gau, M.D. PMID- 15888994 TI - Mammogram myths and facts. The truth about breast cancer screening. PMID- 15888995 TI - Obesity and weight gain increase risk of kidney stones. PMID- 15888996 TI - Daytime naps may benefit some older adults. PMID- 15888998 TI - Common drug interactions. What to watch for. PMID- 15888997 TI - Long-term consumption of red meat increases colon cancer risk. PMID- 15889000 TI - Nausea and vomiting. When queasiness indicates problems. PMID- 15889001 TI - Core strengthening. Improve your balance and stability. PMID- 15889002 TI - Do fiber supplements affect absorption of medicines? PMID- 15889003 TI - Can you tell me more about topical treatments for skin cancer? PMID- 15889004 TI - Does wearing sunscreen prevent me from getting enough vitamin D? PMID- 15889005 TI - Summary of notifiable diseases--United States, 2003. AB - The Summary of Notifiable Diseases--United States, 2003 contains the official statistics, in tabular and graphic form, for the reported occurrence of nationally notifiable diseases in the United States for 2003. Unless otherwise noted, the data are final totals for 2003 reported as of June 30, 2004. These statistics are collected and compiled from reports sent by state health departments to the National Notifiable Diseases Surveillance System (NNDSS), which is operated by CDC in collaboration with the Council of State and Territorial Epidemiologists (CSTE). The Summary is available at http://www.cdc.gov/mmwr/summary.html. This site also includes publications from past years. PMID- 15889006 TI - Positive test results for acute hepatitis A virus infection among persons with no recent history of acute hepatitis--United States, 2002-2004. AB - Hepatitis A is a nationally reportable condition, and the surveillance case definition includes both clinical criteria and serologic confirmation. State health departments and CDC have investigated persons with positive serologic tests for acute hepatitis A virus (HAV) infection (i.e., IgM anti-HAV) whose illness was not consistent with the clinical criteria of the hepatitis A case definition. Test results indicating acute HAV infection among persons who do not have clinical or epidemiologic features consistent with hepatitis A are a concern for state and local health departments because of the need to assess whether contacts need postexposure immunoprophylaxis. This report summarizes results of three such investigations, which suggested that most of the positive tests did not represent recent acute HAV infections. To improve the predictive value of a positive IgM anti-HAV test, clinicians should limit laboratory testing for acute HAV infection to persons with clinical findings typical of hepatitis A or to persons who have been exposed to settings where HAV transmission is suspected. PMID- 15889007 TI - Assessment of epidemiologic capacity in state and territorial health departments- United States, 2004. AB - In November 2001, the Council of State and Territorial Epidemiologists (CSTE) conducted a survey of state and territorial health departments to assess their core epidemiologic capacity. The survey was completed just before distribution of approximately $1 billion in terrorism preparedness and emergency response funds in fiscal year 2002, intended to improve the U.S. public health infrastructure. Results of the 2001 survey, published in 2003, indicated inadequate capacity in six of eight key epidemiology program areas (all except infectious disease and chronic disease) to fully perform the essential public health services most dependent on epidemiology. In 2004, CSTE conducted a follow-up survey that assessed epidemiologic capacity in the United States and its territories in the same eight program areas, estimated the number of additional epidemiologists needed for full performance, and identified education and training needs. This report summarizes the results of that 2004 follow-up survey, which indicated a 26.9% increase from 2001 in the overall number of epidemiologists working in state and territorial health departments, increased capacity in two program areas (i.e., terrorism preparedness and emergency response; maternal and child health) and decreased capacity in six other program areas (i.e., infectious disease, chronic disease, environmental health, injury, occupational health, and oral health). Results also revealed that 28.5% of epidemiologists lacked any formal training or academic coursework in epidemiology. Creation of a strong public health infrastructure fully capable of performing essential services will require additional trained epidemiologists in state and territorial health departments. PMID- 15889008 TI - Terrorism and emergency preparedness in state and territorial public health departments--United States, 2004. AB - After the events of September 11, 2001, federal funding for state public health preparedness programs increased from $67 million in fiscal year (FY) 2001 to approximately $1 billion in FY 2002. These funds were intended to support preparedness for and response to terrorism, infectious disease outbreaks, and other public health threats and emergencies. The Council of State and Territorial Epidemiologists (CSTE) assessed the impact of funding on epidemiologic capacity, including terrorism preparedness and response, in state health departments in November 2001 and again in May 2004, after distribution of an additional $1 billion in FY 2003. This report describes the results of those assessments, which indicated that increased funding for terrorism preparedness and emergency response has rapidly increased the number of epidemiologists and increased capacity for preparedness at the state level. However, despite the increase in epidemiologists, state public health officials estimate that 192 additional epidemiologists, an increase of 45.3%, are needed nationwide to fully staff terrorism preparedness programs. PMID- 15889009 TI - Improvement in local public health preparedness and response capacity--Kansas, 2002-2003. AB - After the terrorist attacks of September 11, 2001, increased funding was provided to federal, state, and local health departments to improve their capacities for terrorism preparedness and emergency response. To evaluate the effect of this funding and to identify priority program areas in Kansas, the Kansas Association of Local Health Departments (KALHD) contracted with the Kansas Health Institute (KHI) to perform an independent assessment of local health department (LHD) preparedness capacity using a CDC assessment tool. This report summarizes the results of two surveys of LHDs and changes in preparedness capacity from 2002 to 2003. The findings indicated a substantial increase in local preparedness capacity, although increases among counties varied widely. Repeated assessments of preparedness using standardized tools can provide useful information to help guide federal, state, and local public health policies and investments. PMID- 15889010 TI - Visual impairment and use of eye-care services and protective eyewear among children--United States, 2002. AB - Visual impairment is an important cause of developmental disability among children. Ocular conditions, if undetected or untreated, can have substantial long-term implications for the quality of life of the child and the family and can place a burden on public health resources. For these reasons, the national health objectives for 2010 now include three vision objectives for children: 1) reducing visual impairment and blindness, 2) increasing the proportion of preschool children who receive vision screening, and 3) increasing the use of protective eyewear in recreational activities and hazardous situations around the home (objective nos. 28-2, 28-4, and 28-9a). When these objectives were published in November 2000, baseline data were available for only one objective, that of reducing visual impairment and blindness among children. To address all three childhood vision objectives, CDC analyzed data from the 2002 National Health Interview Survey (NHIS). This report summarizes the results of that analysis, which indicated that the prevalence of visual impairment and the use of eye-care services were significantly higher among children aged >6 years, compared with younger children, and varied by race/ethnicity and family income. As a result of that analysis, national baselines are now available for all three objectives. Collaborative efforts of all relevant agencies, health professionals, educators, and the public are needed to achieve the national childhood vision objectives for 2010. PMID- 15889011 TI - Outbreak of multidrug-resistant Salmonella typhimurium associated with rodents purchased at retail pet stores--United States, December 2003-October 2004. AB - During 2004, the Minnesota Department of Health (MDH) Public Health Laboratory notified CDC about the isolation of multidrug-resistant Salmonella enterica serotype Typhimurium from ill hamsters from a Minnesota pet distributor. This report describes two of the first identified human cases associated with this outbreak, summarizes the multistate investigation of human S. Typhimurium infections associated with exposure to rodents (e.g., hamsters, mice, and rats) purchased at pet stores, and highlights methods for reducing Salmonella transmission from pet rodents to their owners. This is the first documented salmonellosis outbreak associated with pet rodents. Findings demonstrate that the handling of pet rodents is a potential health risk, especially for children. Public health practitioners should consider pet rodents a potential source of salmonellosis. PMID- 15889012 TI - Social support and health-related quality of life among older adults--Missouri, 2000. AB - Overall health can be influenced by multiple factors, including a person's psychological, behavioral, and social well-being. Studies have demonstrated an association between increased levels of social support and reduced risk for physical disease, mental illness, and mortality. Social support includes real or perceived resources provided by others that enable a person to feel cared for, valued, and part of a network of communication and mutual obligation. Social support can be critical for those older adults who rely on family, friends, or organizations to assist them with daily activities, provide companionship, and care for their well-being. The 1965 Older Americans Act recognized the need for social support by requiring that agencies on aging provide in-home services and group meals to foster social interactions. To examine how social support is related to health-related quality of life (HRQOL), CDC analyzed data from the 2000 Missouri Older Adults Needs Assessment Survey (MOANAS) of adults aged > or =60 years. This report describes the results of that analysis, which indicated that visits with friends or relatives, having close friends for emotional support, and the perception of help being available if sick or disabled were associated with better HRQOL and particularly with better mental health among older adults. Implementing effective prevention programs for older adults and encouraging interventions by agencies on aging can help improve HRQOL among older adults who have little social support. PMID- 15889013 TI - Expression of dominant negative Rho-binding domain of Rho-kinase in organ cultured human eye anterior segments increases aqueous humor outflow. AB - PURPOSE: Based on pharmacological inhibition of activity, a role has been proposed for Rho-kinase in the modulation of aqueous outflow and intraocular pressure (IOP). This study employed a molecular approach to specifically address the role of Rho-kinase in the modulation of aqueous humor outflow. METHODS: Adenoviral vectors expressing green fluorescent protein alone (Ad-GFP) or the dominant negative Rho-binding domain of Rho-kinase and GFP (Ad-DNRK-GFP) were utilized in these experiments. Human and porcine primary trabecular meshwork (TM) cells were infected with 30 MOI (multiplicity of infection) of Ad-GFP alone or with Ad-DNRK-GFP. Changes in cell shape, actomyosin cytoskeletal integrity, and the status of myosin light chain (MLC) phosphorylation were evaluated. The aqueous outflow facility was determined in organ cultured anterior segments of human cadaver eyes infected with 10(7) pfu adenoviral vectors (Ad-GFP or Ad-DNRK GFP) using a constant flow perfusion system. RESULTS: Expression of DNRK resulted in changes in cell shape (cell rounding, cell-cell detachment) and decreased actin stress fiber and focal adhesion staining in TM cells. These cellular changes were associated with substantially reduced myosin light chain phosphorylation. Additionally, organ cultured human eye anterior segments infected with Ad-DNRK-GFP exhibited a significant increase in the outflow facility (80%, n=9) compared to control anterior segments infected with Ad-GFP (5%). CONCLUSIONS: This study demonstrated that specific inhibition of Rho-kinase activity in trabecular meshwork cells led to alterations in cell shape and presumed contractile properties, and we hypothesize that these morphological and contractile events underlie the observed effects of dominant negative Rho-kinase on the aqueous humor outflow facility. These data provide molecular evidence for the hypothesis of Rho-kinase being a potential cellular target involved in the regulation of aqueous humor outflow resistance, with implications for novel glaucoma therapy. PMID- 15889014 TI - Transgenic expression of leukemia inhibitory factor (LIF) blocks normal vascular development but not pathological neovascularization in the eye. AB - PURPOSE: Vascular development in the eye has been described as a complex process involving both vasculogenesis and angiogenesis. Multiple cell types are involved in the process including angioblasts, vascular endothelial cells, astrocytes, pericytes, and Muller glial cells. This suggests that multiple growth factors and cytokines are required to regulate retinal vascular development. Leukemia inhibitory factor (LIF) is a member of the interleukin 6 family of cytokines. LIF is expressed during inflammation and has been reported to affect vascular development in culture; however, its effects in vivo have not been demonstrated. The purpose of this study was to determine how LIF could regulate ocular vascular development. METHODS: We have analyzed ocular vascular development in transgenic mice that express LIF in the ocular lens from embryonic day 11. RESULTS: In transgenic mice, LIF reduced development of embryonic vasculature in the eye, and inhibited retinal vascular development. Inhibition in vivo was independent of vascular endothelial cell growth factor (VEGF) expression. In older transgenic mice, the absence of a retinal vasculature resulted in retinal ischemia and elevated VEGF levels. The upregulation of VEGF resulted in the proliferation of pathological vascular membranes in the vitreous and neovascularization penetrating the retina, which in turn resulted in tractional retinal detachment. CONCLUSIONS: LIF is a potent inhibitor of retinal vascular development. These transgenic mice will be useful as a model of persistent fetal vasculature and to study the mechanism for the development of neovascular membranes in the vitreous and could be used to develop inhibitors of tractional detachment. PMID- 15889015 TI - Regulation of Egr-1, VIP, and Shh mRNA and Egr-1 protein in the mouse retina by light and image quality. AB - PURPOSE: To analyze mRNA expression changes of Egr-1, VIP, and Shh under different light and treatment conditions in mice. The mRNA expression levels of the three genes and additionally the Egr-1 protein expression were compared in form deprived eyes and eyes with normal vision. Moreover, the influence of dark to light and light to dark transitions and of changes in retinal illumination on mRNA levels was investigated. METHODS: Form deprivation of mice was induced by fitting frosted diffusers over one eye and an attentuation matched neutral density (ND) filter over the other eye. To measure the effects of retinal illumination changes on mRNA expression, animals were bilaterally fitted with different ND filters. Semiquantitative real-time RT-PCR was used to measure the mRNA levels and immunohistochemistry was applied to localize and detect Egr-1 protein. RESULTS: The expression levels of both Egr-1 mRNA and protein were reduced in form deprived eyes compared to their fellow eyes after 30 min and 1 h, respectively. Egr-1 mRNA was strikingly upregulated both after dark to light and light to dark transitions, whereas minor changes in retinal illumination by covering the eyes with neutral density filters did not alter Egr-1 mRNA expression. In mice, the mRNA levels of VIP and Shh were not affected by form deprivation, but they were found to be regulated depending on the time of day. CONCLUSIONS: Both Egr-1 mRNA and protein expression levels were strongly regulated by light, especially by transitions between light and darkness. Image contrast may exert an additional influence on mRNA and protein expression of Egr 1, particularly in the cells in the ganglion cell layer and in bipolar cells. PMID- 15889016 TI - Interaction and biophysical properties of human lens Q155* betaB2-crystallin mutant. AB - PURPOSE: Missense mutations in crystallin genes have been identified in autosomal dominant congenital cataracts. A truncation in the CRYBB2 gene (Q155*) has been associated with cerulean cataract, however its effects on biophysical properties have not been reported. We sought to determine the changes in conformation and protein-protein interactions brought about by this mutation. METHODS: Site specific mutations were performed to obtain the Q155* betaB2-crystallin mutant. Protein-protein interactions were screened by a mammalian two-hybrid system assay. Conformational changes were studied with spectroscopy (circular dichroism and fluorescence) and FPLC chromatography. RESULTS: We detected a decrease in protein-protein interactions for the Q155* betaB2-crystallin mutant. The Q155* mutant shows decreased ordered structure and stability but the partially unfolded protein retains some dimer structure. CONCLUSIONS: The Q155* mutation in betaB2 crystallin causes changes in biophysical properties that might contribute to cataract formation. PMID- 15889017 TI - Expression of the p53 family of proteins in central and peripheral human corneal endothelial cells. AB - PURPOSE: To determine the protein and mRNA expression of p53, p63, and p73 in central and peripheral human corneal endothelial cells. Since these proteins are known to be involved in the regulation of cell division, this study seeks information about their influence in regulating cell proliferation in the human corneal endothelium. METHODS: Human donor corneas were separated into central and peripheral sections. The endothelial tissue from these samples was dissected and samples were analyzed for mRNA transcription of p53, transactivating p63 (TAp63), delta N p63 (DeltaNp63), transactivating p73 (TAp73), and delta N p73 (DeltaNp73) via the reverse transcriptase-polymerase chain reaction (RT-PCR). Additional samples were analyzed for p53, p63, and p73 protein expression via SDS-PAGE, western blotting, and immunodetection. Frozen corneal sections were immunostained for p53 and analyzed via fluorescence microscopy. RESULTS: p53 and TAp63 mRNA and protein expression were detected in central and peripheral human corneal endothelium. p53 and TAp63 protein expression were greater in central than in peripheral tissue. DeltaNp63 and all isoforms of p73 were not detected in either central or peripheral corneal endothelium. CONCLUSIONS: p53 is expressed in both peripheral and central human corneal endothelium, although it is more highly expressed in the central endothelium. Similarly, TAp63 is more highly expressed in central rather than in peripheral endothelium. This suggests that the peripheral endothelium may have more potential for cell division than the central endothelium. DeltaNp63, a stem cell marker, was not detected in the corneal endothelium. Neither the TAp73 nor the DeltaNp73 isoforms were detected in either central or peripheral human corneal endothelium. PMID- 15889018 TI - A novel PAX6 gene mutation in a Chinese family with aniridia. AB - PURPOSE: The PAX6 gene mutation in aniridia has been studied in various ethnic patients, but not well studied in the Chinese population. In the present study, we have investigated the PAX6 gene mutation in a Chinese family with congenital aniridia. METHODS: Total genomic DNA was isolated from peripheral blood of three aniridia patients (who also suffered from bilateral congenital cataracts) and two non-carriers in a Chinese family. Fourteen exons of the PAX6 gene were amplified by polymerase chain reaction (PCR). PCR products of each exon were analyzed by single strand conformational polymorphism (SSCP). The PCR products with abnormal SSCP patterns were subcloned and sequenced to identify the mutation. RESULTS: Abnormal SSCP patterns were found in all affected patients but not in non-carrier family members. A novel mutation (c.857delG) in exon 7 was detected by sequencing analysis. This frame shift mutation was predicted to lead to a pre-stop codon in exon 8, and generate a novel 40 amino acid peptide from codon 165. CONCLUSIONS: A novel PAX6 gene mutation was identified in a Chinese aniridia family. This mutation may also contribute to congenital cataracts in these aniridia patients. PMID- 15889019 TI - Overutilization of shoulder magnetic resonance imaging as a diagnostic screening tool in patients with chronic shoulder pain. AB - This was a retrospective study of 101 consecutive patients (104 shoulders) with chronic, atraumatic shoulder pain who were evaluated by a fellowship-trained shoulder specialist. The patients were assessed with a complete history, physical examination, plain radiographs, Simple Shoulder Test, and visual analog scores of pain, shoulder function, and quality of life. Forty-one percent of the shoulders presented with shoulder magnetic resonance imaging (MRI). Data analysis was performed to determine the effect of pre-evaluation MRI on patient treatment and outcome. There were no statistically significant differences in age, sex, affected shoulder, insurance status, mechanism of injury, comorbid conditions, range of motion, treatment, initial outcome parameter assessments, or improvement of outcomes between the patients who had pre-evaluation MRI and those who did not. The majority of the pre-evaluation MRI scans had no impact on the outcome. Routine pre-evaluation with MRI does not appear to have a significant effect on the treatment or outcome and should not be used as a screening tool for atraumatic shoulder pain before a comprehensive clinical evaluation of the shoulder. PMID- 15889020 TI - Arthroscopic tenotomy of the long head of the biceps in the treatment of rotator cuff tears: clinical and radiographic results of 307 cases. AB - The purpose of this study is to evaluate the objective, subjective, and radiographic results of arthroscopic biceps tenotomy in selected patients with rotator cuff tears. Three hundred seven arthroscopic biceps tenotomies were performed in patients with full- thickness rotator cuff tears. Patients were selected for arthroscopic tenotomy if the tear was thought to be irreparable or if the patient was older and not willing to participate in the rehabilitation required after rotator cuff repair. Patients were evaluated clinically and radiographically at a mean of 57 months' follow-up (range, 24-168 months). The mean Constant score increased from 48.4 points preoperatively to 67.6 points postoperatively ( P < .0001). Eighty-seven percent of patients were satisfied or very satisfied with the result. The acromiohumeral interval decreased by a mean of 1.3 mm during the follow-up period and was associated with a longer duration of follow-up ( P < .0001). Preoperatively, 38% of patients had glenohumeral arthritis; postoperatively, 67% of patients had glenohumeral arthritis. Concomitant acromioplasty was statistically associated with better subjective and objective results only in patients with an acromiohumeral distance greater than 6 mm. Fatty infiltration of the rotator cuff musculature had a negative influence on both the functional and radiographic results ( P < .0001). Arthroscopic biceps tenotomy in the treatment of rotator cuff tears in selected patients yields good objective improvement and a high degree of patient satisfaction. Despite these improvements, arthroscopic tenotomy does not appear to alter the progressive radiographic changes that occur with long-standing rotator cuff tears. PMID- 15889021 TI - Computerized assessment of Bankart lesions under tension with magnetic resonance arthrography. AB - Accurate anatomic depiction of Bankart lesions based on magnetic resonance imaging (MRI) is crucial for the treatment of posttraumatic recurrent dislocation of the glenohumeral joint. MR arthrography, the intraarticular injection of dilute gadolinium before MR imaging, improves sensitivity in the detection of shoulder pathology. Abduction and external rotation (ABER) of the shoulder places dynamic stress on the inferior capsular pouch and anterior labroligamentous complex, important structures for anterior shoulder stability. This study sought to determine whether MR arthrography, by use of computerized image analysis, can visualize Bankart lesions better with the shoulder in the neutral position or in ABER. We evaluated 12 shoulders after traumatic anterior dislocation. The MR images were analyzed with image-analyzing software. We compared 3 parameters at 5 levels of the glenoid in the neutral position and in ABER: detachment (the length of detachment between the anterior glenoid rim and the anterior periosteal attachment), displacement (the distance between the anterior glenoid rim and the tip of the displaced labrum), and Bankart area (the area bounded by the detachment line, the displacement line, and the anterior aspect of the Bankart lesion). MR images revealed that Bankart lesions were under tension in ABER and lax and redundant with the shoulder in the neutral position. All 3 parameters were greater in ABER than in the neutral position in all cases (P = .012, P = .0006, and P = .012). Computerized image assessment of MR arthrography with the shoulder in ABER provides excellent visualization and evaluation of Bankart lesions. PMID- 15889022 TI - Radiographic comparison of pegged and keeled glenoid components. AB - Glenoid loosening is one reason for failure of total shoulder arthroplasty. Several factors, including radiographic lucency, have been shown to be associated with glenoid loosening. The purpose of this study was to assess the correlation between glenoid design and immediate radiographic lucency in a prospective randomized clinical trial. Total shoulder arthroplasty was performed in 43 patients over a 2-year period. Twenty-three patients were randomized into the keel group and twenty patients into the pegged group. Postoperative radiographs obtained within 6 weeks of surgery were evaluated by 3 raters to determine glenoid lucency. On a scale from 0 (no lucency) to 5 (gross lucency and component loosening), the rate of lucency was 39% (9/23) in the keeled components, which was significantly higher than the rate of 5% (1/20) observed in the pegged components (P = .026). Patient age, gender, and glenoid size did not significantly affect glenoid component lucency (P > .05). The consistency reliability among raters (Cronbach alpha) was 0.87, and the intertester reliability was 0.87. Pegged glenoid components have less radiographic lucency when compared with keeled glenoid components in the immediate postoperative period. PMID- 15889023 TI - Humeral windows in revision shoulder arthroplasty. AB - Currently, there are no published results of patients in whom a window in the humeral cortex had been created to facilitate revision of a failed shoulder arthroplasty. Therefore, we reviewed the results of patients who underwent creation of a humeral window in the course of a revision shoulder arthroplasty to determine the complications and rate of healing. Twenty patients underwent humeral osteotomy to remove a well-fixed humeral component during the course of revision shoulder arthroplasty at our institution between January 1, 1987, and December 31, 2002. There were 17 cemented humeral components and 3 fully textured, press-fitted components. Of the shoulders, 16 had an anterior window, 3 had a medial window, and 1 had combined anterior and medial windows. There were 4 intraoperative fractures: 3 in the humeral shaft and 1 in the greater tuberosity. At the most recent radiographic follow-up at a mean of 3.3 years (range, 3-176 months), 17 of 20 windows had healed. Among 3 patients with limited radiographic follow-up (1, 3, and 4 months), 2 were healing and 1 did not demonstrate radiographic signs of healing. There were no cases of window malunion. No humeral components have developed clinical loosening. The data from this study suggest that humeral windows may facilitate controlled removal of well-fixed humeral components with a high rate of union. Further evolution of this technique may result in a lower fracture rate. PMID- 15889024 TI - Free vascularized corticoperiosteal bone graft for the treatment of persistent nonunion of the clavicle. AB - Radiation-induced necrosis or infection can result in a persistent nonunion of the clavicle. We describe 3 cases in which a free vascularized corticoperiosteal flap was harvested from the medial aspect of the femoral condyle and transferred to the fractured clavicle by use of microvascular anastomoses. Healing was confirmed by tomography in all 3 patients. The function of the upper extremity was remarkably improved at final follow-up. Vascularized free corticoperiosteal bone flaps are an option in clavicle fracture treatment. The technique of periosteal grafting is ideally suited to chronic nonunions with poor chances of healing on their own. Rapid subperiosteal new bone and improved local blood flow serve to correct many of those changes attributed to necrosis of bone. PMID- 15889025 TI - Electromyographic evaluation after primary repair of massive rotator cuff tears. AB - Anatomic dissections have suggested that mobilization of the supraspinatus and infraspinatus tendons greater than 1 cm may cause suprascapular nerve injury. This study evaluated rotator cuff function by use of electromyographic (EMG) analysis after primary repair of massive tears in 9 patients. All subjects had massive rotator cuff tears involving at least the entire supraspinatus and entire infraspinatus tendons. Rotator cuff advancement averaged 2.5 cm (range, 2.0-3.5 cm). No EMG evidence of suprascapular nerve injury was found postoperatively. The UCLA shoulder score improved from a preoperative mean of 11 to a postoperative mean of 28. There was 1 excellent, 6 good, 1 fair, and 1 poor result. The poor result was due to a documented failed repair. The EMG results suggest that mobilization and advancement of the rotator cuff up to 3.5 cm during primary repair may be performed safely without as much risk to the suprascapular nerve as anatomic studies would suggest. PMID- 15889026 TI - Intraobserver and interobserver reliability for the strength test in the Constant Murley shoulder assessment. AB - This study evaluates the standardized strength test in the Constant-Murley shoulder assessment of adults with healthy shoulders in a randomized, single blind design. The following questions were to be answered: (1) Can the spring balance and a digital dynamometer both yield the same result? (2) What is the intraobserver and interobserver reliability of the strength test? (3) Is the strength test sensitive to change in technique or affected by calculation with mean or maximum values? Ten subjects were included in a comparison of the Handyscale (digital dynamometer) and the mechanical spring balance for concurrent validity, resulting in intraclass correlation coefficient values ranging from 0.96 to 0.99. For intraobserver and interobserver reliability, 2 observers tested 20 subjects with the Handyscale and retested them after 2 weeks. Regardless of technique during testing, this resulted in almost perfect agreement (intraclass correlation coefficient range, 0.89-0.98). The digital dynamometer can replace the conventional spring balance. The standardized strength test in the Constant Murley shoulder assessment is reliable in young subjects with healthy shoulders, independent of technique or whether calculated with mean or maximum values. PMID- 15889027 TI - Normalization of the Constant score. AB - The strength of the normal shoulder may differ by gender and deteriorate with age. Thus, the Constant score may also decrease in absolute value while still reflecting a normal score. To account for age- and gender-related differences, normal results for this scale must be determined across a population of patients without shoulder disease. Patients presenting for evaluation of nonshoulder conditions participated. A subjective questionnaire was completed. Range of motion and strength were measured. This analysis includes the data of 441 patients. The mean Constant score for men was significantly greater than that for women in each age group ( P < .05). Significant age-related differences were noted in each group ( P < .05). Normative values for the Constant score based on age and gender were determined. The adjusted score represents the gender- and age matched function of the shoulder and is useful in the evaluation of shoulder outcomes. PMID- 15889028 TI - Measurement of baseline shoulder function in subjects receiving workers' compensation versus noncompensated subjects. AB - The purpose of this study was to ascertain whether a measurable difference existed in normative scores for the American Shoulder and Elbow Surgeons (ASES) questionnaire between subjects who had an active workers' compensation claim (WC) with no known shoulder injury and subjects without a compensation claim (non-WC). Subjects with non-shoulder-related orthopaedic injuries were recruited from a suburban orthopaedic sports medicine clinic and an urban occupational medicine clinic. They were asked to complete a composite questionnaire that consisted of demographic information and the ASES questionnaire. There were no significant differences in the ASES scores between subject groups. There were significant differences between subject groups with regard to work hours (P = .0001), work demands (P = .0001), and tobacco use (P = .0001). Subject group was also significantly associated with education level (P = .0001), marital status (P = .0001), work demands (P = .0001), gender (P = .0001), and sports participation (P = .0314). The ASES score was significantly affected by marital status (P = .0476), sports participation (P = .0008), and age (P = .0129). PMID- 15889029 TI - The effect of arthroscopic sectioning of the lateral ligament complex of the elbow on posterolateral rotatory stability. AB - This study evaluates the relative roles of the radial collateral ligament, the lateral ulnar collateral ligament, and the overlying musculature in posterolateral rotatory instability of the elbow. Fourteen cadaveric upper limbs underwent sequential arthroscopic sectioning of the lateral collateral ligament complex. After sectioning, arthroscopic and fluoroscopic evaluation of a lateral pivot shift test was done. Minimal instability was noted after the first section, but no difference between radial collateral or lateral ulnar collateral ligament sectioning was found. A greater degree of instability was seen between the first and second cut ( P = .0001), but no significant difference was seen between sectioning the 2 groups ( P = .61). Complete instability occurred only after sectioning the overlying musculature. On the basis of this study, injury to both the radial collateral and lateral ulnar collateral ligaments is necessary to cause significant posterolateral rotatory instability of the elbow. Furthermore, the overlying musculature plays an important role in overall stability. PMID- 15889030 TI - Patient-oriented functional outcome after repair of distal biceps tendon ruptures using a single-incision technique. AB - Using a patient-oriented outcome questionnaire, in addition to standard outcome measures, we sought to determine the outcome of patients who had repair of a complete rupture of the distal biceps tendon via a single anterior incision technique with suture anchors. We identified 62 patients who were treated operatively by a single surgeon over an 8-year period for a diagnosis of complete rupture of the distal biceps tendon. Of the patients, 9 could not be located for final follow-up, and 53 of 62 (85%) participated in the study. All patients were men, and their mean age was 42 years. All repairs were performed via a single anterior incision by use of 2 suture anchors in the bicipital tuberosity on an outpatient basis. There were 4 complications (4/53 [7.5%]): 1 wound infection, 2 transient paresthesias in the lateral cutaneous nerve distribution, and 1 posterior interosseous nerve palsy that resolved in 6 weeks (no reoperations). There were no reruptures, and no patient lost more than 5 degrees of elbow flexion- extension or forearm rotation. All patients completed the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire. The mean DASH score was 8.2 +/- 11.6 (95% CI, 5.2 to 11.9), which was similar to the mean DASH score in population controls of 6.2 (DASH User Manual). The strengths of our study include the consistent surgical technique by a single surgeon, the large number of patients, and the use of a patient-oriented outcome measure. Distal biceps tendon rupture repair via a single-incision technique with suture anchors was effective in restoring injured arms to normal, as measured by limb-specific patient oriented measures, with minimal morbidity and a low complication rate. PMID- 15889031 TI - Herbert screw fixation of capitellar fractures. AB - From 1993 to 2003, 6 patients with displaced fractures of the humeral capitellum were treated by open reduction and internal fixation of the capitellar fragments with Herbert bone screws. By use of the criteria of Grantham et al, there were 2 type II-A fractures, 1 type II-B fracture, 1 type II-C fracture, and 2 type III-A fractures. A lateral approach was used in 4 patients and a posterior approach with olecranon osteotomy in 2. The elbows were immobilized postoperatively for 4 to 28 days (mean, 13.5 days). We evaluated the range of motion, stability, and pain using the criteria of Grantham et al. The follow-up period ranged from 2.5 to 9.3 years (mean, 5.6 years). All patients had a stable, pain-free elbow with good range of motion at follow-up. All fractures healed, and there was no evidence of avascular necrosis or degenerative change. PMID- 15889032 TI - The drop sign, a radiographic warning sign of elbow instability. AB - Persistent instability or redislocation is uncommon but of significant concern in treating elbow dislocations. Finding an objective, static radiographic sign that might correlate with the presence of instability was the purpose of this study. Pre- and post-reduction radiographs of 10 consecutive simple and complete adult elbow dislocations were reviewed and compared with radiographs of 20 consecutive adult elbows without any trauma history. A statistically significant measured increase in static ulnohumeral distance was noted on the routine unstressed post reduction lateral radiographs of patients sustaining dislocation. We have termed this increased distance the drop sign. It differs from the radiographic ulnohumeral separation noted during O'Driscoll's test for posterolateral rotary instability, which is present only with axial compression. The drop sign becomes concerning only if persistent after the first reduction radiograph and may be a warning sign of the presence of instability. PMID- 15889033 TI - Biomechanical evaluation after five and ten millimeter anterior glenohumeral capsulorrhaphy using a novel shoulder model of increased laxity. AB - Anterior instability of the shoulder is classically treated with a capsulolabral repair, but in cases of capsular redundancy, shortening or shifting of the capsule is added. This study compared glenohumeral translations in intact shoulders after rotational stretching of the capsule and after progressive increasing of anterior-inferior capsular shifts. Seven cadaveric shoulders were mounted on a custom biomechanical testing apparatus. Rotational range of motion and glenohumeral translations were measured. To create laxity, the shoulders were rotationally stretched an additional 30% from the intact rotational range of motion about the axis of the humerus in external and internal rotation. Anterior inferior capsular shifts of 5 and 10 mm were performed. Rotational stretching of the shoulder capsule created anterior laxity. A 5 mm capsular shift was ineffective, but a 10 mm shift restored anterior and total anteroposterior translation to the intact condition. PMID- 15889034 TI - The arthroscopic view of the glenohumeral ligaments compared with anatomy: fold or fact? AB - In a morphologic cadaveric study with observational arthroscopy in living subjects, we tried to resolve the contradiction in the literature with regard to the nature of the glenohumeral ligaments and the difference in observation of the folds during arthroscopic and open surgery. Observation of morphology and functional anatomy of the glenohumeral capsule was performed in 200 non-embalmed cadavers through open dissection (100 specimens) and by arthroscopy (50 specimens) or both (50 specimens), as well as in 100 living subjects undergoing shoulder arthroscopy. In the resting arm position, folds and bands can be observed on the inside of the anteroinferior capsule. When the arm is moved into full abduction and external rotation, however, all bands progressively disappear from sight. The bands generally observed in the shoulder capsule during arthroscopy appear at the site of histologic reinforcements of the capsule but are not the capsular ligaments themselves, as they seem to disappear in certain positions of the humerus. Arthroscopically, it is, therefore, not possible to discern the exact limits of these ligaments. This may give rise to a certain amount of confusion when comparing clinical with anatomic and physiologic studies. On the other hand, their presence or absence in arthroscopic surgery might be of clinical relevance in evaluating capsular tension. PMID- 15889035 TI - Osteoid osteoma of the scapula associated with synovitis of the shoulder. PMID- 15889036 TI - Gonococcal osteomyelitis of the humeral head: a case report. PMID- 15889037 TI - Posttraumatic locked dislocation of congenital pseudarthrosis of the clavicle. PMID- 15889038 TI - Dislocation of the glenoid fossa. PMID- 15889039 TI - Isolated fracture of the trochlea: a case report. PMID- 15889041 TI - Lag signs. PMID- 15889042 TI - Epilepsy and apoptosis pathways. AB - Epilepsy is a common, chronic neurologic disorder characterized by recurrent unprovoked seizures. Experimental modeling and clinical neuroimaging of patients has shown that certain seizures are capable of causing neuronal death. Such brain injury may contribute to epileptogenesis, impairments in cognitive function or the epilepsy phenotype. Research into cell death after seizures has identified the induction of the molecular machinery of apoptosis. Here, the authors review the clinical and experimental evidence for apoptotic cell death pathway function in the wake of seizure activity. We summarize work showing intrinsic (mitochondrial) and extrinsic (death receptor) apoptotic pathway function after seizures, activation of the caspase and Bcl-2 families of cell death modulators and the acute and chronic neuropathologic impact of intervening in these molecular cascades. Finally, we describe evolving data on nonlethal roles for these proteins in neuronal restructuring and cell excitability that have implications for shaping the epilepsy phenotype. This review highlights the work to date on apoptosis pathway signaling during seizure-induced neuronal death and epileptogenesis, and speculates on how emerging roles in brain remodeling and excitability have enriched the number of therapeutic strategies for protection against seizure-damage and epileptogenesis. PMID- 15889043 TI - MR perfusion and diffusion in acute ischemic stroke: human gray and white matter have different thresholds for infarction. AB - It is thought that gray and white matter (GM and WM) have different perfusion and diffusion thresholds for cerebral infarction in humans. We sought to determine these thresholds with voxel-by-voxel, tissue-specific analysis of co-registered acute and follow-up magnetic resonance (MR) perfusion- and diffusion-weighted imaging. Quantitative cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT), and apparent diffusion coefficient (ADC) maps were analyzed from nine acute stroke patients (imaging acquired within 6 h of onset). The average values of each measure were calculated for GM and WM in normally perfused tissue, the region of recovered tissue and in the final infarct. Perfusion and diffusion thresholds for infarction were determined on a patient-by patient basis in GM and WM separately by selecting thresholds with equal sensitivities and specificities. Gray matter has higher thresholds for infarction than WM (P<0.009) for CBF (20.0 mL/100 g min in GM and 12.3 mL/100 g min in WM), CBV (2.4 mL/100 g in GM and 1.7 mL/100 g in WM), and ADC (786 x 10(-6) mm(2)/s in GM and 708 x 10(-6) mm(2)/s in WM). The MTT threshold for infarction in GM is lower (P=0.014) than for WM (6.8 secs in GM and 7.1 secs in WM). A single common threshold applied to both tissues overestimates tissue at risk in WM and underestimates tissue at risk in GM. This study suggests that tissue-specific analysis of perfusion and diffusion imaging is required to accurately predict tissue at risk of infarction in acute ischemic stroke. PMID- 15889044 TI - Delayed treatment with monoclonal antibody IN-1 1 week after stroke results in recovery of function and corticorubral plasticity in adult rats. AB - Neuronal death due to ischemic stroke results in permanent deficits in sensory, language, and motor functions. The growth-restrictive environment of the adult central nervous system (CNS) is an obstacle to functional recovery after stroke and other CNS injuries. In this regard, Nogo-A is a potent neurite growth inhibitory protein known to restrict neuronal plasticity in adults. Previously, we have found that treatment with monoclonal antibody (mAb) IN-1 to neutralize Nogo-A immediately after stroke enhanced motor cortico-efferent plasticity and recovery of skilled forelimb function in rats. However, immediate treatment for stroke is often not clinically feasible. Thus, the present study was undertaken to determine whether cortico-efferent plasticity and functional recovery would occur if treatment with mAb IN-1 was delayed 1 week after stroke. Adult rats were trained on a forelimb-reaching task, and the middle cerebral artery was occluded to induce focal cerebral ischemia to the forelimb sensorimotor cortex. After 1 week, animals received mAb IN-1 treatment, control antibody, or no treatment, and were tested for 9 more weeks. To assess cortico-efferent plasticity, the sensorimotor cortex opposite the stroke lesion was injected with an anterograde neuroanatomical tracer. Behavioral analysis demonstrated a recovery of skilled forelimb function, and anatomical studies revealed neuroplasticity at the level of the red nucleus in animals treated with mAb IN-1, thus demonstrating the efficacy of this treatment even if administered 1 week after stroke. PMID- 15889045 TI - A novel method to derive separate gray and white matter cerebral blood flow measures from MR imaging of acute ischemic stroke patients. AB - Perfusion-weighted imaging (PWI) measures can predict tissue outcome in acute ischemic stroke. Accuracy might be improved if differential tissue susceptibility to ischemia is considered. We present a novel voxel-by-voxel analysis to characterize cerebral blood flow (CBF) separately in gray (GM) and white matter (WM). Ten patients were scanned with inversion-recovery spin-echo EPI (IRSEPI), diffusion-weighted imaging (DWI), PWI<6 h from onset and fluid attenuated inversion-recovery (FLAIR) at 30 days. Image processing included coregistration to PWI, automatic segmentation of IRSEPI into GM, WM and CSF and semiautomatic segmentation of DWI/FLAIR to derive the acute and 30-day lesions. Five tissue compartments were defined: (1) 'Core' (abnormal acutely and at 30 days), (2) 'Growth' (or 'infarcted penumbra', abnormal only at 30 days), (3) 'Reversed' (abnormal acutely but normal at 30 days), (4) 'MTT-Delayed ' (tissue with delayed mean transit time but not part of the acute or 30-day lesion), and (5) 'Normal' brain. Cerebral blood flow in GM and WM of each compartment was obtained from quantitative maps. Gray matter and WM mean CBF in the growth region differed by 5.5 mL/100 g min (P=0.015). Mean CBF also differed significantly within normal and MTT-Delayed compartments. The difference in the reversed region approached statistical significance. In core, GM and WM CBF did not differ. The results suggest separate ischemic thresholds for GM and WM in stroke penumbra. PMID- 15889046 TI - Allele variations in the OCA2 gene (pink-eyed-dilution locus) are associated with genetic susceptibility to melanoma. AB - The occuloalbinism 2 (OCA2) gene, localized at 15q11, encodes a melanosomal transmembrane protein that is involved in the most common form of human occulo cutaneous albinism, a human genetic disorder characterized by fair pigmentation and susceptibility to skin cancer. We wondered whether allele variations at this locus could influence susceptibility to malignant melanoma (MM). In all, 10 intragenic single-nucleotide polymorphisms (SNPs) were genotyped in 113 patients with melanomas and in 105 Caucasian control subjects with no personal or family history of skin cancer. By comparing allelic distribution between cases and controls, we show that MM and OCA2 are associated (p value=0.030 after correction for multiple testing). Then, a recently developed strategy, the 'combination test' enabled us to show that a combination formed by two SNPs was most strongly associated to MM, suggesting a possible interaction between intragenic SNPs. In addition, the role of OCA2 on MM risk was also detected using a logistic model taking into account the presence of variants of the melanocortin 1 receptor gene (MC1R, a key pigmentation gene) and all pigmentation characteristics as melanoma risk factors. Our data demonstrate that a second pigmentation gene, in addition to MC1R, is involved in genetic susceptibility to melanoma. PMID- 15889047 TI - Stargardt's disease and retinitis pigmentosa: different phenotypic presentations in the same family. PMID- 15889048 TI - Genome-wide analysis of human kinases in clathrin- and caveolae/raft-mediated endocytosis. AB - Endocytosis is a key cellular process, encompassing different entry routes and endocytic compartments. To what extent endocytosis is subjected to high-order regulation by the cellular signalling machinery remains unclear. Using high throughput RNA interference and automated image analysis, we explored the function of human kinases in two principal types of endocytosis: clathrin- and caveolae/raft-mediated endocytosis. We monitored this through infection of vesicular stomatitis virus, simian virus 40 and transferrin trafficking, and also through cell proliferation and apoptosis assays. Here we show that a high number of kinases are involved in endocytosis, and that each endocytic route is regulated by a specific kinase subset. Notably, one group of kinases exerted opposite effects on the two endocytic routes, suggesting coordinate regulation. Our analysis demonstrates that signalling functions such as those controlling cell adhesion, growth and proliferation, are built into the machinery of endocytosis to a much higher degree than previously recognized. PMID- 15889050 TI - Blair's failure. PMID- 15889051 TI - 'Refusal to share' leaves agency struggling to monitor bird flu. PMID- 15889052 TI - Nations spar over erosion of nuclear treaty. PMID- 15889053 TI - Polio fight falters as Yemen and Java report fresh cases. PMID- 15889054 TI - Early martian visitors are caught on camera. PMID- 15889055 TI - Competition boosts bid to find human genes. PMID- 15889056 TI - Wanted: scientists to shape Europe's future research policy. PMID- 15889057 TI - Cleaner skies leave global warming forecasts uncertain. PMID- 15889060 TI - Chinese clinical trials: consenting adults? Not necessarily... PMID- 15889061 TI - Spanish astronomy: rising star. PMID- 15889062 TI - Seeing clearly is not necessarily believing. PMID- 15889063 TI - Two-stage drug approval would reduce the risks. PMID- 15889064 TI - Universities should foster neglected-disease work. PMID- 15889070 TI - Natural symmetry. PMID- 15889071 TI - Immunology: insulin trigger for diabetes. PMID- 15889072 TI - High-energy physics: an emptier emptiness? PMID- 15889075 TI - Planetary science: magnetic impact craters. PMID- 15889074 TI - Developmental biology: asymmetrical threat averted. PMID- 15889076 TI - Sensory physiology: brainless eyes. PMID- 15889077 TI - Granular media: information propagation. PMID- 15889078 TI - Obituary: Stanley J. Korsmeyer (1950-2005). PMID- 15889080 TI - Robotics: self-reproducing machines. AB - Self-reproduction is central to biological life for long-term sustainability and evolutionary adaptation. Although these traits would also be desirable in many engineered systems, the principles of self-reproduction have not been exploited in machine design. Here we create simple machines that act as autonomous modular robots and are capable of physical self-reproduction using a set of cubes. PMID- 15889081 TI - Botany: a record-breaking pollen catapult. AB - The release of stored elastic energy often drives rapid movements in animal systems, and plant components employing this mechanism should be able to move with similar speed. Here we describe how the flower stamens of the bunchberry dogwood (Cornus canadensis) rely on this principle to catapult pollen into the air as the flower opens explosively. Our high-speed video observations show that the flower opens in less than 0.5 ms--to our knowledge, the fastest movement so far recorded in a plant. PMID- 15889082 TI - Retinoic acid signalling links left-right asymmetric patterning and bilaterally symmetric somitogenesis in the zebrafish embryo. AB - During embryogenesis, cells are spatially patterned as a result of highly coordinated and stereotyped morphogenetic events. In the vertebrate embryo, information on laterality is conveyed to the node, and subsequently to the lateral plate mesoderm, by a complex cascade of epigenetic and genetic events, eventually leading to a left-right asymmetric body plan. At the same time, the paraxial mesoderm is patterned along the anterior-posterior axis in metameric units, or somites, in a bilaterally symmetric fashion. Here we characterize a cascade of laterality information in the zebrafish embryo and show that blocking the early steps of this cascade (before it reaches the lateral plate mesoderm) results in random left-right asymmetric somitogenesis. We also uncover a mechanism mediated by retinoic acid signalling that is crucial in buffering the influence of the flow of laterality information on the left-right progression of somite formation, and thus in ensuring bilaterally symmetric somitogenesis. PMID- 15889083 TI - FGF-induced vesicular release of Sonic hedgehog and retinoic acid in leftward nodal flow is critical for left-right determination. AB - The precise specification of left-right asymmetry is an essential process for patterning internal organs in vertebrates. In mouse embryonic development, the symmetry-breaking process in left-right determination is initiated by a leftward extraembryonic fluid flow on the surface of the ventral node. However, it is not known whether the signal transduction mechanism of this flow is chemical or mechanical. Here we show that fibroblast growth factor (FGF) signalling triggers secretion of membrane-sheathed objects 0.3-5 microm in diameter termed 'nodal vesicular parcels' (NVPs) that carry Sonic hedgehog and retinoic acid. These NVPs are transported leftward by the fluid flow and eventually fragment close to the left wall of the ventral node. The silencing effects of the FGF-receptor inhibitor SU5402 on NVP secretion and on a downstream rise in Ca2+ were sufficiently reversed by exogenous Sonic hedgehog peptide or retinoic acid, suggesting that FGF-triggered surface accumulation of cargo morphogens may be essential for launching NVPs. Thus, we propose that NVP flow is a new mode of extracellular transport that forms a left-right gradient of morphogens. PMID- 15889084 TI - A link between prompt optical and prompt gamma-ray emission in gamma-ray bursts. AB - The prompt optical emission that arrives with the gamma-rays from a cosmic gamma ray burst (GRB) is a signature of the engine powering the burst, the properties of the ultra-relativistic ejecta of the explosion, and the ejecta's interactions with the surroundings. Until now, only GRB 990123 had been detected at optical wavelengths during the burst phase. Its prompt optical emission was variable and uncorrelated with the prompt gamma-ray emission, suggesting that the optical emission was generated by a reverse shock arising from the ejecta's collision with surrounding material. Here we report prompt optical emission from GRB 041219a. It is variable and correlated with the prompt gamma-rays, indicating a common origin for the optical light and the gamma-rays. Within the context of the standard fireball model of GRBs, we attribute this new optical component to internal shocks driven into the burst ejecta by variations of the inner engine. The correlated optical emission is a direct probe of the jet isolated from the medium. The timing of the uncorrelated optical emission is strongly dependent on the nature of the medium. PMID- 15889085 TI - An infrared flash contemporaneous with the gamma-rays of GRB 041219a. AB - The explosion that results in a cosmic gamma-ray burst (GRB) is thought to produce emission from two physical processes: the central engine gives rise to the high-energy emission of the burst through internal shocking, and the subsequent interaction of the flow with the external environment produces long wavelength afterglows. Although observations of afterglows continue to refine our understanding of GRB progenitors and relativistic shocks, gamma-ray observations alone have not yielded a clear picture of the origin of the prompt emission nor details of the central engine. Only one concurrent visible-light transient has been found and it was associated with emission from an external shock. Here we report the discovery of infrared emission contemporaneous with a GRB, beginning 7.2 minutes after the onset of GRB 041219a (ref. 8). We acquired 21 images during the active phase of the burst, yielding early multi-colour observations. Our analysis of the initial infrared pulse suggests an origin consistent with internal shocks. PMID- 15889086 TI - Albedo of the south pole on Mars determined by topographic forcing of atmosphere dynamics. AB - The nature of the martian south polar cap has remained enigmatic since the first spacecraft observations. In particular, the presence of a perennial carbon dioxide ice cap, the formation of a vast area of black 'slab ice' known as the Cryptic region and the asymmetric springtime retreat of the cap have eluded explanation. Here we present observations and climate modelling that indicate the south pole of Mars is characterized by two distinct regional climates that are the result of dynamical forcing by the largest southern impact basins, Argyre and Hellas. The style of surface frost deposition is controlled by these regional climates. In the cold and stormy conditions that exist poleward of 60 degrees S and extend 180 degrees in longitude west from the Mountains of Mitchel (approximately 30 degrees W), surface frost accumulation is dominated by precipitation. In the opposite hemisphere, the polar atmosphere is relatively warm and clear and frost accumulation is dominated by direct vapour deposition. It is the differences in these deposition styles that determine the cap albedo. PMID- 15889087 TI - Friction enhances elasticity in granular solids. AB - For years, engineers have used elastic and plastic models to describe the properties of granular solids, such as sand piles and grains in silos. However, there are theoretical and experimental results that challenge this approach. Specifically, it has been claimed that stress in granular solids propagates in a manner described by wave-like (hyperbolic) equations, rather than the elliptic equations of static elasticity. Here we report numerical simulations of the response of a two-dimensional granular slab to an external load, revealing that both approaches are valid--albeit on different length scales. For small systems that can be considered mesoscopic on the scale of the grains, a hyperbolic-like, strongly anisotropic response is expected. However, in large systems (those typically considered by engineers), the response is closer to that predicted by traditional isotropic elasticity models. Static friction, often ignored in simple models, plays a key role: it increases the elastic range and renders the response more isotropic, even beyond this range. PMID- 15889088 TI - Nonlinear elasticity in biological gels. AB - The mechanical properties of soft biological tissues are essential to their physiological function and cannot easily be duplicated by synthetic materials. Unlike simple polymer gels, many biological materials--including blood vessels, mesentery tissue, lung parenchyma, cornea and blood clots--stiffen as they are strained, thereby preventing large deformations that could threaten tissue integrity. The molecular structures and design principles responsible for this nonlinear elasticity are unknown. Here we report a molecular theory that accounts for strain-stiffening in a range of molecularly distinct gels formed from cytoskeletal and extracellular proteins and that reveals universal stress-strain relations at low to intermediate strains. The input to this theory is the force extension curve for individual semi-flexible filaments and the assumptions that biological networks composed of these filaments are homogeneous, isotropic, and that they strain uniformly. This theory shows that systems of filamentous proteins arranged in an open crosslinked mesh invariably stiffen at low strains without requiring a specific architecture or multiple elements with different intrinsic stiffness. PMID- 15889089 TI - Plate-wide stress relaxation explains European Palaeocene basin inversions. AB - During Late Cretaceous and Cenozoic times, many Palaeozoic and Mesozoic rifts and basin structures in the interior of the European continent underwent several phases of inversion (the process of shortening a previously extensional basin). The main phases occurred during the Late Cretaceous and Middle Palaeocene, and have been previously explained by pulses of compression, mainly from the Alpine orogen. Here we show that the main phases differed both in structural style and cause. The Cretaceous phase was characterized by narrow uplift zones, reverse activation of faults, crustal shortening, and the formation of asymmetric marginal troughs. In contrast, the Middle Palaeocene phase was characterized by dome-like uplift of a wider area with only mild fault movements, and formation of more distal and shallow marginal troughs. A simple flexural model explains how domal, secondary inversion follows inevitably from primary, convergence-related inversion on relaxation of the in-plane tectonic stress. The onset of relaxation inversions was plate-wide and simultaneous, and may have been triggered by stress changes caused by elevation of the North Atlantic lithosphere by the Iceland plume or the drop in the north-south convergence rate between Africa and Europe. PMID- 15889090 TI - Palaeomagnetism of the Vredefort meteorite crater and implications for craters on Mars. AB - Magnetic surveys of the martian surface have revealed significantly lower magnetic field intensities over the gigantic impact craters Hellas and Argyre than over surrounding regions. The reduced fields are commonly attributed to pressure demagnetization caused by shock waves generated during meteorite impact, in the absence of a significant ambient magnetic field. Lower than average magnetic field intensities are also observed above the Vredefort meteorite crater in South Africa, yet here we show that the rocks in this crater possess much higher magnetic intensities than equivalent lithologies found elsewhere on Earth. We find that palaeomagnetic directions of these strongly magnetized rocks are randomly oriented, with vector directions changing over centimetre length scales. Moreover, the magnetite grains contributing to the magnetic remanence crystallized during impact, which directly relates the randomization and intensification to the impact event. The strong and randomly oriented magnetization vectors effectively cancel out when summed over the whole crater. Seen from high altitudes, as for martian craters, the magnetic field appears much lower than that of neighbouring terranes, implying that magnetic anomalies of meteorite craters cannot be used as evidence for the absence of the planet's internally generated magnetic field at the time of impact. PMID- 15889091 TI - Advanced optics in a jellyfish eye. AB - Cubozoans, or box jellyfish, differ from all other cnidarians by an active fish like behaviour and an elaborate sensory apparatus. Each of the four sides of the animal carries a conspicuous sensory club (the rhopalium), which has evolved into a bizarre cluster of different eyes. Two of the eyes on each rhopalium have long been known to resemble eyes of higher animals, but the function and performance of these eyes have remained unknown. Here we show that box-jellyfish lenses contain a finely tuned refractive index gradient producing nearly aberration-free imaging. This demonstrates that even simple animals have been able to evolve the sophisticated visual optics previously known only from a few advanced bilaterian phyla. However, the position of the retina does not coincide with the sharp image, leading to very wide and complex receptive fields in individual photoreceptors. We argue that this may be useful in eyes serving a single visual task. The findings indicate that tailoring of complex receptive fields might have been one of the original driving forces in the evolution of animal lenses. PMID- 15889092 TI - The flight paths of honeybees recruited by the waggle dance. AB - In the 'dance language' of honeybees, the dancer generates a specific, coded message that describes the direction and distance from the hive of a new food source, and this message is displaced in both space and time from the dancer's discovery of that source. Karl von Frisch concluded that bees 'recruited' by this dance used the information encoded in it to guide them directly to the remote food source, and this Nobel Prize-winning discovery revealed the most sophisticated example of non-primate communication that we know of. In spite of some initial scepticism, almost all biologists are now convinced that von Frisch was correct, but what has hitherto been lacking is a quantitative description of how effectively recruits translate the code in the dance into flight to their destinations. Using harmonic radar to record the actual flight paths of recruited bees, we now provide that description. PMID- 15889093 TI - The origin of bursts and heavy tails in human dynamics. AB - The dynamics of many social, technological and economic phenomena are driven by individual human actions, turning the quantitative understanding of human behaviour into a central question of modern science. Current models of human dynamics, used from risk assessment to communications, assume that human actions are randomly distributed in time and thus well approximated by Poisson processes. In contrast, there is increasing evidence that the timing of many human activities, ranging from communication to entertainment and work patterns, follow non-Poisson statistics, characterized by bursts of rapidly occurring events separated by long periods of inactivity. Here I show that the bursty nature of human behaviour is a consequence of a decision-based queuing process: when individuals execute tasks based on some perceived priority, the timing of the tasks will be heavy tailed, with most tasks being rapidly executed, whereas a few experience very long waiting times. In contrast, random or priority blind execution is well approximated by uniform inter-event statistics. These finding have important implications, ranging from resource management to service allocation, in both communications and retail. PMID- 15889094 TI - Retinoic acid coordinates somitogenesis and left-right patterning in vertebrate embryos. AB - A striking feature of the body plan of a majority of animals is bilateral symmetry. Almost nothing is known about the mechanisms controlling the symmetrical arrangement of the left and right body sides during development. Here we report that blocking the production of retinoic acid (RA) in chicken embryos leads to a desynchronization of somite formation between the two embryonic sides, demonstrated by a shortened left segmented region. This defect is linked to a loss of coordination of the segmentation clock oscillations. The lateralization of this defect led us to investigate the relation between somitogenesis and the left-right asymmetry machinery in RA-deficient embryos. Reversal of the situs in chick or mouse embryos lacking RA results in a reversal of the somitogenesis laterality defect. Our data indicate that RA is important in buffering the lateralizing influence of the left-right machinery, thus permitting synchronization of the development of the two embryonic sides. PMID- 15889095 TI - Prime role for an insulin epitope in the development of type 1 diabetes in NOD mice. AB - A fundamental question about the pathogenesis of spontaneous autoimmune diabetes is whether there are primary autoantigens. For type 1 diabetes it is clear that multiple islet molecules are the target of autoimmunity in man and animal models. It is not clear whether any of the target molecules are essential for the destruction of islet beta cells. Here we show that the proinsulin/insulin molecules have a sequence that is a primary target of the autoimmunity that causes diabetes of the non-obese diabetic (NOD) mouse. We created insulin 1 and insulin 2 gene knockouts combined with a mutated proinsulin transgene (in which residue 16 on the B chain was changed to alanine) in NOD mice. This mutation abrogated the T-cell stimulation of a series of the major insulin autoreactive NOD T-cell clones. Female mice with only the altered insulin did not develop insulin autoantibodies, insulitis or autoimmune diabetes, in contrast with mice containing at least one copy of the native insulin gene. We suggest that proinsulin is a primary autoantigen of the NOD mouse, and speculate that organ restricted autoimmune disorders with marked major histocompatibility complex (MHC) restriction of disease are likely to have specific primary autoantigens. PMID- 15889096 TI - Expanded T cells from pancreatic lymph nodes of type 1 diabetic subjects recognize an insulin epitope. AB - In autoimmune type 1 diabetes, pathogenic T lymphocytes are associated with the specific destruction of insulin-producing beta-islet cells. Identification of the autoantigens involved in triggering this process is a central question. Here we examined T cells from pancreatic draining lymph nodes, the site of islet-cell specific self-antigen presentation. We cloned single T cells in a non-biased manner from pancreatic draining lymph nodes of subjects with type 1 diabetes and from non-diabetic controls. A high degree of T-cell clonal expansion was observed in pancreatic lymph nodes from long-term diabetic patients but not from control subjects. The oligoclonally expanded T cells from diabetic subjects with DR4, a susceptibility allele for type 1 diabetes, recognized the insulin A 1-15 epitope restricted by DR4. These results identify insulin-reactive, clonally expanded T cells from the site of autoinflammatory drainage in long-term type 1 diabetics, indicating that insulin may indeed be the target antigen causing autoimmune diabetes. PMID- 15889097 TI - Enhancement of cellular memory by reducing stochastic transitions. AB - On induction of cell differentiation, distinct cell phenotypes are encoded by complex genetic networks. These networks can prevent the reversion of established phenotypes even in the presence of significant fluctuations. Here we explore the key parameters that determine the stability of cellular memory by using the yeast galactose-signalling network as a model system. This network contains multiple nested feedback loops. Of the two positive feedback loops, only the loop mediated by the cytoplasmic signal transducer Gal3p is able to generate two stable expression states with a persistent memory of previous galactose consumption states. The parallel loop mediated by the galactose transporter Gal2p only increases the expression difference between the two states. A negative feedback through the inhibitor Gal80p reduces the strength of the core positive feedback. Despite this, a constitutive increase in the Gal80p concentration tunes the system from having destabilized memory to having persistent memory. A model reveals that fluctuations are trapped more efficiently at higher Gal80p concentrations. Indeed, the rate at which single cells randomly switch back and forth between expression states was reduced. These observations provide a quantitative understanding of the stability and reversibility of cellular differentiation states. PMID- 15889099 TI - Hot from the vent. PMID- 15889100 TI - PCR: replicating success. PMID- 15889101 TI - Amplifying the signal. PMID- 15889102 TI - Photocopiers for DNA. PMID- 15889103 TI - Simplifying the probe set. PMID- 15889105 TI - Generous advice. PMID- 15889106 TI - The staff dreams are made of. PMID- 15889108 TI - Scientists and societies. Postdoc power. PMID- 15889111 TI - Putting related individual studies into childhood obesity intervention studies. PMID- 15889112 TI - Development and modification of child food preferences and eating patterns: behavior genetics strategies. AB - Behavioral genetics (BG) designs can offer useful strategies for studying the development of child food preferences and eating patterns. This review summarizes BG designs that tested familial influences on child eating behavior and implicated both genetic and home environmental factors. A range of BG strategies, including family and pseudo-family designs, classic twins designs, discordant sibling designs, cotwin control designs, and high-risk designs, have provided information on child development that could not have been obtained from the analysis of singletons. BG designs can provide can powerful tools for testing environmental theories of child nutrition and, potentially, for better understanding between-child variability in response to dietary interventions for overweight. The term BG may misleadingly imply only the classic twin design or just heritability estimation; BG strategies can be adapted creatively to address a range of questions concerning the development of child appetite and eating regulation. PMID- 15889113 TI - BMI from 3-6 y of age is predicted by TV viewing and physical activity, not diet. AB - OBJECTIVE: To investigate whether, diet, physical activity, sedentary behavior or television (TV) viewing predicted body mass index (BMI) among 3-7-y-old children. DESIGN: A triethnic cohort of 3-4-y-old children was followed for 3 y from 1986 to 1989. MEASUREMENTS: BMI was assessed at the beginning and end of each measurement year. Heart rate monitoring and observation were used to assess physical activity. Diet (calories, % calories from fat and carbohydrate), sedentary behavior and TV viewing were assessed by direct observation in each year. A repeated measures regression analysis with year as a factor and BMI at the end of each year as dependent variables was run. Nonsignificant variables were removed in a stepwise backward deletion process and significant interactions graphed. RESULTS: The interactions between minutes of TV viewing per hour and study year and minutes of physical activity per hour and study year were significant (P<0.05). There were also significant main effects for TV viewing, physical activity and BMI from the beginning of the study. The model accounted for 65% of the variance in BMI across the three study years. Plotting the significant interactions demonstrated that physical activity was positively associated with BMI in year 1, and negatively associated in years 2 and 3 with a stronger negative relationship in year 3 than 2. TV viewing became positively associated with BMI during the third study year. CONCLUSION: Physical activity and TV viewing were the only significant predictors (other than baseline BMI) of BMI among a triethnic cohort of 3-4-y-old children followed for 3 y with both physical activity (negatively associated) and TV viewing (positively associated) becoming stronger predictors as the children aged. It appears that 6 or 7 y is a critical age when TV viewing and physical activity may affect BMI. Therefore, focusing on reducing time spent watching television and increasing time spent in physical activity may be successful means of preventing obesity among this age group. PMID- 15889114 TI - In situ lipolytic regulation in subjects born small for gestational age. AB - OBJECTIVE: Subjects born small for gestational age (SGA) who are prone to develop insulin resistance in adulthood display an abnormal development pattern of the adipose tissue during fetal and postnatal life. Since the lipolytic activity of the adipose tissue is critical in the development of insulin resistance, the purpose of this study was to investigate whether SGA itself might affect lipolysis regulation. STUDY DESIGN: We studied the effect of catecholamines, by local injection of isoproterenol, and the effect of insulin, using two-step infusion at 8 and 40 mU/m2/min, on the in situ lipolysis of the subcutaneous abdominal adipose tissue of 23 subjects born SGA and 23 born appropriate for gestational age (AGA), using the microdialysis technique. RESULTS: Under isoproterenol infusion, the increase in dialysate glycerol concentration was significantly 1.5-fold higher in the SGA than in the AGA group (P=0.02) and induced a 20% increase in the plasma FFA concentration (P=0.04), whereas no significant increase was observed in the AGA group. The antilipolytic action of insulin on dialysate glycerol concentration was similar in both groups throughout the insulin infusion. CONCLUSION: Subjects born SGA demonstrated a hyperlipolytic reactivity to catecholamines, which might be regarded as an additional deleterious component of the insulin resistance associated with SGA. In contrast, being born SGA does not directly affect the antilipolytic action of insulin, showing that it does not play a major role in causing the long-term metabolic complications associated with reduced fetal growth. PMID- 15889115 TI - Insulin resistance and impaired glucose tolerance in obese children and adolescents referred to a tertiary-care center in Israel. AB - OBJECTIVE: To establish the prevalence of insulin resistance and impaired glucose tolerance (IGT) and their determinants in a cohort of obese children and adolescents. METHODS: A retrospective design was used. The study group included 234 patients with a body mass index (BMI) greater than the 95th percentile for age and gender and 22 patients with a BMI between the 85th and 95th percentile for age and gender referred for evaluation to a major tertiary-care center in Israel. Ages ranged from 5 to 22 y. Estimates of insulin resistance (homeostatic model assessment (HOMA-IR)); insulin sensitivity (ratio of fasting glucose (GF) to fasting insulin (IF) (GF/IF), the quantitative insulin sensitivity check index (QUICKI)), and pancreatic beta-cell function (HOMA-derived beta-cell function (HOMA %B)) were derived from fasting measurements. An oral glucose tolerance test (OGTT) was performed in 192 patients to determine the presence of IGT. RESULTS: Insulin resistance was detected in 81.2% of the patients, IGT in 13.5%, and silent diabetes in one adolescent girl. Only two patients with IGT also had impaired fasting glucose (IFG). The prevalence of IGT was higher in adolescents than prepubertal children (14.7 vs 8.6%). GF/IF and QUICKI decreased significantly during puberty (P<0.005), whereas HOMA-IR and HOMA %B did not. Insulin resistance and insulin sensitivity indexes were not associated with ethnicity, presence of acanthosis nigricans or family history of type 2 diabetes. Patients with obesity complications had lower insulin sensitivity indexes than those without (P=0.05). Compared with subjects with normal glucose tolerance (NGT), patients with IGT had significantly higher fasting blood glucose (85.9+/ 6.5 vs 89.2+/-10.6 mg/dl, P<0.05), higher 2-h post-OGGT insulin levels (101.2+/ 74.0 vs 207.6+/-129.7 microU/ml, P<0.001), a lower QUICKI (0.323+/-0.031 vs 0.309+/-0.022, P<0.05), and higher fasting triglyceride levels (117.4+/-53.1 vs 156.9+/-68.9, P=0.002). However, several of the fasting indexes except QUICKI failed to predict IGT. There was no difference between the group with IGT and the group with NGT in fasting insulin, HOMA-IR, HOMA %B or the male-to-female ratio, age, BMI-SDS, presence of acanthosis nigricans, ethnicity, and family history of type 2 diabetes. CONCLUSIONS: Insulin resistance is highly prevalent in obese children and adolescents. The onset of IGT is associated with the development of severe hyperinsulinemia as there are no predictive cutpoint values of insulin resistance or insulin sensitivity indexes for IGT, and neither fasting blood glucose nor insulin levels nor HOMA-IR or HOMA %B are effective screening tools; an OGTT is required in all subjects at high risk. Longitudinal studies are needed to identify the metabolic precursors and the natural history of the development of type 2 diabetes in these patients. PMID- 15889116 TI - Ten-year trends for fatness in Northern Irish adolescents: the Young Hearts Projects--repeat cross-sectional study. AB - OBJECTIVE: To examine secular trends in measures of fatness over a 10-y period (the 1990s) in Northern Irish schoolchildren. DESIGN: Repeat cross-sectional surveys. SUBJECTS: In total, 12- and 15-y-old children randomly selected from post-primary schools. A total of 1015 children studied between 1989 and 1990, and 2017 studied between 1999 and 2001. MEASUREMENTS: The same study methods were applied in both surveys. Body mass index (BMI) was calculated from height and weight, and percentage body fat (%BF) was derived from skinfold measurements at four sites. Dietary intake was assessed using a diet history method and habitual physical activity (PA) was estimated from a self-report questionnaire. RESULTS: Increases were seen in both height and weight in all age-sex groups. The overall prevalence of overweight/obesity increased from 15.0 to 19.6%, but 12-y-old girls contributed most to this increase (15.9-26.3%), with a modest increase observed also in 15-y-old boys. Increases in mean BMI (19.2-20.3 kg/m2) and mean %BF (25.8 27.1%) were seen in 12-y-old girls, with no significant changes in any of the other subgroups. Mean energy intake increased in girls but not in boys, while mean PA score decreased in 12-y-old girls, but was unchanged in the other three groups. All age-sex groups showed substantial increases in the sugar intake while fat intake increased in girls and decreased in boys. CONCLUSION: Increases in indices of fatness were seen among school-age children in Northern Ireland during the 1990s. Trends differed between age-sex groups with the largest changes seen in 12-y-old girls. It remains to be seen whether the large increase in overweight/obesity in this group tracks into adulthood, with consequences for chronic disease incidence in women. PMID- 15889117 TI - Changes in physical activity explain paradoxical relationship between baseline physical activity and adiposity changes in adolescent girls: the FLVS II study. AB - OBJECTIVE: To investigate in adolescents relationships between habitual physical activity (PA) and changes in several indicators of adiposity. DESIGN: Longitudinal population-based study. The Fleurbaix-Laventie Ville Sante II (FLVS II) study in Northern France. SUBJECTS: A total of 222 boys and 214 girls aged 8 18 y, not obese at inclusion. MEASUREMENTS: PA (Modifiable Activity Questionnaire) and adiposity (body mass index (kg/m2), percent body fat by bioimpedance analysis, sum of four skinfolds, waist circumference) were assessed in 1999 and 2001. Adiposity indicators adjusted for age and pubertal stage were compared between groups of baseline PA and between groups of PA changes, separately by sex. RESULTS: At baseline, PA was not associated with adiposity indicators in both sexes. In girls only, being in the group with the highest age relative level of moderate PA at baseline predicted the highest adiposity gain from baseline to follow-up, for all indicators. After adjustment for baseline values, all adiposity indicators were higher at follow-up in girls who had decreased their relative level of moderate PA during follow-up. In boys, changes in adiposity during follow-up were not significantly different across groups of PA changes. However, the sum of skinfolds at follow-up, adjusted for baseline value, tended to be higher in those who decreased their vigorous PA level and lower in those who increased it. CONCLUSION: In adolescent girls, a decrease in PA rather than inactivity per se may result in increased adiposity over time. PMID- 15889118 TI - Percent body fat measured by BIA and DEXA in obese, African-American adolescent girls. AB - OBJECTIVE: To compare the estimation of body fat between bioelectrical impedance analysis (BIA) and dual energy X-ray absorptiometry (DEXA) in overweight, African American female adolescents. SUBJECTS: In total, 54 African-American adolescent female subjects were recruited for Study 1. Each adolescent's body mass index was greater than the 85th percentile and their average body fat was 45% according to DEXA. A total of 26 African-American adolescent female subjects were available for Study 2, and had an average body fat of 26% according to DEXA. MEASUREMENTS: Percent body fat was measured by DEXA and BIA. Seven different BIA equations were tested. Both sets of data were analyzed using Bland-Altman regression analyses, utilizing percent body fat measured by DEXA as the criterion. RESULTS: The Kushner equation provided estimates that were unaffected by body fat in both studies. Estimates were unbiased when applied to the exclusively overweight sample and biased when utilized with the separate sample of normal weight and obese girls. The remaining equations were biased, provided inconsistent estimates across body weight, or were biased and provided inconsistent estimates. Ethnicity specific and ethnicity-combined equations performed similarly in the obese sample, but became more disparate when applied to a sample encompassing a wider body weight range. The limits of agreement between all BIA equations and the DEXA estimates ranged from 6 to 9%. CONCLUSION: The study suggests that the Kushner BIA equation is appropriate for use with African-American female adolescents across the weight spectrum, while the majority of BIA equations underestimated percent body fat as body fat increased. PMID- 15889119 TI - Cutaneous temperature measurements in men with penile prostheses: a comparison study. AB - To evaluate and compare the cutaneous temperature of the penis in normal men, those with erectile dysfunction (ED), those with semirigid penile prostheses (SRPPs), and those with inflatable penile prostheses (IPPs), and those before and after trimix injection to create a penile erection. A total of 68 patients were evaluated. Five patient groups were identified, including men with normal erectile function, with ED, with SRPPs, with IPPs, and following intracavernosal injection of trimix solution. Cutaneous glans temperature increased significantly by more than 2.2 degrees C in the trimix-injected group compared with all other groups (P<0.001). Using cutaneous temperature measurements of the penis, patients with SRPPs had significantly lower cutaneous glans temperatures than normals (P<0.02), those in the ED group (P<0.04), and those in the IPP-deflated group (P<0.01). The mean temperature difference was 1.44+/-0.40 degrees C. Using cutaneous temperature measurements of the penis, men with SRPPs have a colder glans as compared with men with normal erectile function, ED, IPPs, and those who have received an injection of trimix. Men with normal erectile function, ED, and IPPs did not have significant cutaneous temperature differences. PMID- 15889120 TI - Secondhand tobacco smoke impairs neurogenic and endothelium-dependent relaxation of rabbit corpus cavernosum smooth muscle: improvement with chronic oral administration of L-arginine. AB - The first goal of this study was to examine the effect of secondhand smoking on neurogenic, endothelium- and cGMP-dependent relaxant responses of rabbit corpus cavernosum smooth muscle. Our second goal was to determine whether such an effect can be prevented by oral administration of L-arginine. Male New Zealand rabbits were divided into control, chronic passive cigarette smoking and L-arginine treatment groups. Relaxant or contractile responses in isolated corpus cavernosum smooth muscle strips were determined by using in vitro muscle technique. There was no significant difference in the relaxant response of the strips to papaverine, sodium nitroprusside and contractile response to KCl among the groups. Relaxant responses to acetylcholine and electrical field stimulation and contractile response to phenylephrine were significantly decreased in the strips of the smoking group than that of the control group. The impaired relaxations of strips were markedly improved by treatment of L-arginine, but the contractile responses to phenylephrine were not affected. These data indicate that secondhand smoking may impair both neurogenic and endothelium-dependent relaxation of corpus cavernosum smooth muscle, and may contribute to the etiology of impotence. Chronic dietary supplementation with L-arginine offsets the impairment of neurogenic and endothelial relaxation. Therefore, we suggest that secondhand smoking exposure to cigarette produces selective impairment of neurogenic and endothelium-dependent relaxation of corpus cavernosum smooth muscle via a mechanism related to the decreased production and/or availability of nitric oxide. PMID- 15889121 TI - Correlation between LUTS (AUA-SS) and erectile dysfunction (SHIM) in an age matched racially diverse male population: data from the Prostate Cancer Awareness Week (PCAW). AB - The relationship between lower urinary tract symptoms (LUTS) due to benign prostatic hyperplasia and sexual health in men participating in a national multicenter screening program was studied. A total of 12 679 men were screened for prostate cancer in the year 2003. Of these, 6641 men had completed both the American Urological Association Symptom Score (AUA-SS) and the Sexual Health Inventory for Men (SHIM) questionnaires. We assessed the apparent effect of comorbidities (ischemic heart disease, hypertension, hypercholesteremia and diabetes), smoking habits and testosterone level on the overall sexual health. Age and race were also assessed as factors affecting the SHIM score. We used a general linear multivariable regression analysis to express the effect of these variables on the sexual health in these men adjusting for the apparent effect of LUTS. The mean and median age of the population was 58.4 +/- 9.8 and 58 y, respectively. The median AUA-SS was 4/25 (mean=5.7 +/- 5.3) and SHIM score was 19/25 (mean=16.3 +/- 5.9). Of the men, 4948 (75%) were Caucasian and 1154 (17%) were from African-American racial origin. A high AUA-SS appears to have a negative effect on the overall sexual health (P<0.05) after adjusting for all other confounding factors. As expected, age showed a significant inverse correlation with SHIM score (P<0.05). Caucasian men on average appear to have a significantly higher SHIM score by 6.5 points when compared to African-American men after adjusting for age, comorbidities, smoking habits, and AUA-SS (P<0.05). However, with increasing age, the difference in SHIM score diminishes between the two groups. Further, smoking and comorbidities were strong predictors of poor sexual health performance. Interestingly, hypogonadism (testosterone <300 ng/dl) was not a significant risk factor (P=0.104) when adjusting for all other variables. Nonetheless, in a univariate analysis, testosterone levels significantly correlated with reported SHIM scores (P<0.05). The overall sexual health in aging men is substantially affected not only by age, but by the severity of their urinary symptoms after adjusting for the most common known risk factors, suggesting perhaps a common underlying pathophysiology. Moreover, race appears to constitute another neglected potential risk factor, which should be investigated further in future studies. PMID- 15889122 TI - Initial validation of a novel rat model of vasculogenic erectile dysfunction with generalized atherosclerosis. AB - Although rats have been widely used in evaluating various causes of vasculogenic erectile dysfunction (VED), the atherosclerotic rat model has seldom been tried probably due to its inherent tolerance to a cholesterol diet. To enhance endothelial sensitivity to cholesterol diet, we tested the effects of transient interruption of nitric oxide synthase on atherogenesis induced by cholesterol diet in a rat model. Rats with atherosclerosis (AS group) received 1% cholesterol diet for 6 weeks. During the initial 2 weeks, they drank water that contained N(G)-nitro-L-arginine methyl ester (L-NAME) (3 mg/ml). After 6 weeks, we carried out histologic and hemodynamic evaluation to confirm pelvic atherosclerosis and erectile dysfunction, respectively, and the results were compared with those of cholesterol only (Chol) group and normal control (C) group. Compared to the C or Chol group, the mean intima/media (I/M) of the internal pudendal artery, which contributes approximately 70% of the total resistance of the penile vasculature, was markedly increased by the treatment (1.82+/-0.25 vs 0.77+/-0.13, P<0.05). Correspondingly, significantly diminished erectile function was observed. Combined treatment for 2 weeks elicited early atherosclerotic changes in proximal arteries and erectile impairment and further 4 weeks of cholesterol diet spread overt atherosclerosis to the periphery. The Chol group showed no arterial pathology, although they showed mild VED. A correlation study showed that atherosclerosis of the distal artery was better correlated with erectile dysfunction than the proximal artery. Based on these results, our study demonstrates that combination treatment of cholesterol diet with L-NAME would be used as a rapid, effective protocol of developing atherosclerotic rat model of VED. PMID- 15889123 TI - Recovery of erection after pelvic urologic surgery: our experience. AB - The incidence of erectile dysfunction (ED) in patients undergoing pelvic urologic surgery, the efficacy and tolerability of vardenafil-based rehabilitative treatment as first option in these patients, the role of spontaneous erection (SE) as a possible positive predictive factor to erection recovery after such treatment, and the role of second-line therapies in those nonresponders are evaluated. All the patients undergoing pelvic urologic surgery at our Institution between November 2002 and December 2003 were considered. Preoperative erectile function (EF) was evaluated by using the abridged five-item version of the International Index of Erectile Function (IIEF5) questionnaire. Study population was divided into separate groups considering grade of preoperative EF, nerve sparing (NS) surgery and type of procedure (radical prostatectomy, radical cystectomy (RC) or nerve and seminal sparing cystectomy). In total, 86 patients were evaluated. After 6 months, an increase in mean IIEF5 score of 12.9 points was found in those who had undergone a bilateral NSRP after vardenafil therapy, of 8.0 points in those who had undergone unilateral NSRP, of 11.3 in those who had undergone NSRC and of 11.5 in nerve and seminal sparing cistectomies. A better vardenafil response was found in patients with SE+(P<0.001). Among those vardenafil notresponders, 13 were treated by using intracavernous injections, one by vacuum device and three with penile prosthesis implant. In conclusion, in our experience, vardenafil showed to be well tolerated and effective for recovery of EF in patients undergoing pelvic urologic surgery. This drug was particularly effective for those with a normal preoperative EF undergoing an NS procedure. Of course, it should be recognized that the absence of a control group in the study represents an important limitation. However, based on the data from the literature, there is a strong belief that such an approach will lead to an earlier recovery of EF than without rehabilitative treatment. PMID- 15889124 TI - Evaluation of a progressive treatment program for erectile dysfunction in patients with diabetes mellitus. AB - The aim was to evaluate the effectiveness of a progressive program, starting with simple methods and, when not effective, moving to more complex methods, to treat erectile dysfunction (ED) in patients with diabetes mellitus. A total of 284 diabetic patients with ED entered into a 6-phase program starting with sildenafil citrate (Viagra). Those with contraindications, side effects, or negative response (erection insufficient for vaginal penetration) were switched to the vacuum erection device (VED), and then progressively (for failures) to intracavernous injection (ICI), sildenafil citrate+ICI, ICI+VED, and penile prosthesis. Patients were followed for 2 y. Of the 284 patients 276 patients were eligible for sildenafil citrate and 147 (53.3%) responded positively, but 25 (9.1%) patients stopped it soon due to adverse effects. Of 162 patients (129 nonresponders, eight noneligible for the sildenafil and 25 patients who dropped out due to adverse effects), treated with VED, 114 (70.4%) responded well, however, only 19 (11.7%) patients agreed to continue its use. Of the remaining 143 patients (nonresponders, noneligible for the previously mentioned treatments and patients who dropped out due to adverse effects), 103/143 (72%) responded to ICI, 27/40 (67.5%) to sildenafil+ICI, and 9/13 (69.2%) to ICI+VED. Four patients received a penile implant. At the 2 y follow-up, 81 of 284 patients who entered the study (28.5%) were still responding to sildenafil, seven (2.5%) to VED, 113 (39.8%) to ICI, 24 (8.5%) to sildenafil+ICI, two (0.7%) to ICI+VED; 15 (5.3%) had a penile implant. In all 17 (6%) patients reported spontaneous erections, 11 (3.9%) stopped the treatment due to family reasons and 14 (4.9%) failed the treatment. In conclusion, the progressive treatment program for ED seems to be very effective for diabetic patients, yielded a complete response for short-term and 91.2% rate of success at the end of 2 y follow-up. PMID- 15889125 TI - Testosterone therapy in women: a review. AB - Female sexual dysfunction is a complex problem with multiple overlapping etiologies. Androgens play an important role in healthy female sexual function, especially in stimulating sexual interest and in maintaining desire. There are a multitude of reasons why women can have low androgen levels with the most common reasons being age, oophorectomy and the use of oral estrogens. Symptoms of androgen insufficiency include absent or greatly diminished sexual motivation and/or desire, that is, libido, persistent unexplainable fatigue or lack of energy, and a lack of sense of well being. Although there is no androgen preparation that has been specifically approved by the FDA for the treatment of Women's Sexual Interest/Desire Disorder or for the treatment of androgen insufficiency in women, androgen therapy has been used off-label to treat low libido and sexual dysfunction in women for over 40 y. Most clinical trials in postmenopausal women with loss of libido have demonstrated that the addition of testosterone to estrogen significantly improved multiple facets of sexual functioning including libido and sexual desire, arousal, frequency and satisfaction. In controlled clinical trials of up to 2 y duration of testosterone therapy, women receiving androgen therapy tolerated androgen administration well and demonstrated no serious side effects. The results of these trials suggest that testosterone therapy in the low-dose regimens is efficacious for the treatment of Women's Sexual Interest and Desire Disorder in postmenopausal women who are adequately estrogenized. Based on the evidence of current studies, it is reasonable to consider testosterone therapy for a symptomatic androgen-deficient woman with Women's Sexual Interest and Desire Disorder. PMID- 15889126 TI - Commentary on androgen deficiency in women and the FDA advisory board's recent decision to request more safety data. AB - Editor's note: The FDA's decision regarding Intrinsa has potential widespread implications for the medical treatment of female sexual dysfunction (FSD). Dr Andre Guay is a world-renowned authority on the topic of female sexuality and FSD. Accompanying Dr Spark's perspective in issue 2, this perspective will shed important light on the topic and offer insight into the past, present and future of FSD. PMID- 15889127 TI - Testosterone replacement therapy and the risk of prostate cancer: a perspective view. AB - The topic of testosterone (T) and prostate cancer is timely and provocative. The Institute of Medicine report on testosterone and aging suggested that knowledge regarding T-replacement therapy in aging men was inadequate, urging a cautious approach to T replacement in this population. The Food and Drug Administration appears to support this cautionary stance. Counter arguments by Rhoden and Morgentaler in the New England Journal of Medicine suggest that T-replacement therapy in aging men may be safe under controlled settings. This perspective continues the debate, and offers a unique perspective from the vantage point of a uro-oncologist. PMID- 15889128 TI - Family-based association analysis implicates IL-4 in susceptibility to Kawasaki disease. AB - Several compelling lines of evidence suggest an important influence of genetic variation in susceptibility to Kawasaki disease (KD), an acute vasculitis that causes coronary artery aneurysms in children. We performed a family-based genotyping study to test for association between KD and 58 genes involved in cardiovascular disease and inflammation. By analysis of a cohort of 209 KD trios using the transmission disequilibrium test, we documented the asymmetric transmission of five alleles including the interleukin-4 (IL-4) C(-589)T allele (P=0.03). Asymmetric transmission of the IL-4 C(-589)T was replicated in a second, independent cohort of 60 trios (P=0.05, combined P=0.002). Haplotypes of alleles in IL-4, colony-stimulating factor 2 (CSF2), IL-13, and transcription factor 7 (TCF7), all located in the interleukin gene cluster on 5q31, were also asymmetrically transmitted. The reported associations of KD with atopic dermatitis and allergy, elevated serum IgE levels, eosinophilia, and increased circulating numbers of monocyte/macrophages expressing the low-affinity IgE receptor (FCepsilonR2) may be related to effects of IL-4. Thus, the largest family-based genotyping study of KD patients to date suggests that genetic variation in the IL-4 gene, or regions linked to IL-4, plays an important role in KD pathogenesis and disease susceptibility. PMID- 15889129 TI - Possible relations between the polymorphisms of the cytokines IL-19, IL-20 and IL 24 and plaque-type psoriasis. AB - The aim of present study was to elucidate the role of the interleukin (IL)-24 gene in predicting risk for plaque-type psoriasis and to describe the linkage disequilibrium (LD) pattern emerging from the genes of IL-19, IL-20 and IL-24. Genes encoding IL-19, IL-20 and IL-24 locate in the region q32 of chromosome 1. The association between the single-nucleotide polymorphisms (SNPs) or haplotypes of the IL-24 gene and the susceptibility of psoriasis was not found. However, a significant protective effect of the combined haplotype CAAAC of IL-20 and IL-24 genes against plaque-type psoriasis was established (OR 0.154). Protective effect against psoriasis was also observed with haplotype TGGGT (OR 0.591) and haplotype CGAGT (OR 0.457). Performing a comprehensive analysis using the data regarding SNPs of IL-24 gene together with the previously published data regarding IL-19 and IL-20 SNPs, we identified two haplotype blocks within the region q32 of chromosome 1. The main result of the present study is that while the IL-19/IL-20 extended haplotype CACCGGAA is a significant susceptibility factor for psoriasis (previous study), IL-20/IL-24 haplotypes CAAAC, TGGGT and CGAGT have a significant protective effect. Nevertheless, family-based studies are required to confirm the impact of IL-19, IL-20 and IL-24 genes in the genetic predisposition for psoriasis. PMID- 15889130 TI - Evidence for natural selection in the HAVCR1 gene: high degree of amino-acid variability in the mucin domain of human HAVCR1 protein. AB - The family of genes encoding T-cell immunoglobulin and mucin-domain containing proteins (Tim), which are cell-surface molecules expressed in CD4(+) T helper cells, has important roles in the immune system. Here, we report three unusual patterns of genetic variation in the human hepatitis A virus cellular receptor 1 gene (HAVCR1) that are similar to patterns observed in major histocompatibility complex loci. First, levels of polymorphism in exon 4 of HAVCR1 were exceptionally high in humans (nucleotide diversity (pi)=45.45 x 10(-4)). Second, nonsynonymous substitutions and insertion/deletion variants were more frequent than synonymous substitutions in that exon (10 out of 12 variants). The rate of the mean number of nucleotide substitutions at nonsynonymous sites to synonymous sites at HAVCR1-exon 4 is >1 (P(A)/P(S)=1.92 and pi(A)/pi(S)=2.23). Third, levels of divergence among human, chimp, and gorilla sequences were unusually high in HAVCR1-exon 4 sequences. These features suggest that patterns of variation in HAVCR1 have been shaped by both positive and balancing natural selection in the course of primate evolution. Evidence that the effects of natural selection are largely restricted to the mucin domain of HAVCR1 suggests that this region may be of particular evolutionary and epidemiological interest. PMID- 15889131 TI - Perinatal care in Arizona 1950-2002: a study of the positive impact of technology, regionalization and the Arizona perinatal trust. PMID- 15889132 TI - Inhaled NO and markers of oxidant injury in infants with respiratory failure. AB - BACKGROUND: Inhaled nitric oxide (iNO) is an effective adjunct in the treatment of infants with respiratory failure. Although there are clear benefits to this therapy, potential toxicity could result from reactive nitrosylated species. OBJECTIVE: To evaluate whether iNO therapy is associated with increased serum markers of oxidative stress. DESIGN/METHOD: Multiple markers were prospectively evaluated in the serum of term infants with severe respiratory failure treated with iNO for 1 to 72 hours. These were compared to those of patients exposed to greater than 80% oxygen for more than 6 hours and room air controls. RESULTS: After 24 hours of exposure, the iNO-treated infants had increased serum lipid hydroperoxides (LPO), protein carbonyls and nitrotyrosine residues as well as increased serum total glutathione (GSH) content. The increase in LPO peaked at 24 hours and correlated with the cumulative dose of iNO whereas other markers did not. The presence of chronic lung disease (CLD) did not correlate with serum markers of oxidative injury. CONCLUSIONS: In term infants with respiratory failure, prolonged iNO exposure is associated with a transient increase in markers of oxidative stress, but this finding does not appear to predict the development of CLD. PMID- 15889133 TI - Interneonatal intensive care unit variation in growth rates and feeding practices in healthy moderately premature infants. AB - BACKGROUND: Variation in care and outcomes of very low birth weight infants (VLBW) in neonatal intensive care units (NICU) has been widely reported in the past decade. Less is known about care provided to healthy premature infants born between 30 and 35 weeks gestational age (GA). We have previously reported inter NICU variation in discharge (D/C) timing and achievement of maturational milestones in this population. OBJECTIVE: To compare inter-NICU growth outcomes and feeding practices in healthy, moderately premature infants. METHODS: Records of 450 infants, 30 to 35 weeks gestation, without medical or surgical complications, and consecutively discharged from 15 Massachusetts NICUs (nine Level II and six Level III) were reviewed. Final analyses included 382 infants with hospital length of stay >6 days (d). RESULTS: GA at birth and birth weight (BW) were 33.2 weeks (SD 1.2) and 2024 g (389). Mean Z-score decreased 0.67z (0.37) from birth to D/C. Weight loss from birth to 7 d averaged 4.0%. Mean growth velocity from 7 d to D/C was 13.3 g/k/d (5.2) with net growth velocity of 5.5 g/k/d (5.6). Mean net growth velocity ranged from 0.1 to 8.4 g/k/d (p<0.001) among study NICUs. Time of initiation, rate of advancement and caloric density of feedings also varied significantly between NICUs. CONCLUSION: Mean NICU growth velocity of healthy, moderately premature infants did not achieve in utero growth standards. There was significant inter-NICU variation in growth outcomes and feeding practices. Further study is needed to identify practices associated with better growth in this healthy moderately premature infant population. PMID- 15889134 TI - In vivo gene transfer to the rat retina using herpes simplex virus type 1 (HSV-1) based amplicon vectors. AB - The purpose of our study was to evaluate the transduction profiles of herpes simplex virus type 1 (HSV-1)-based amplicon vectors following subretinal injection in the rat. Two amplicon vectors were tested, pHy-CMVGFP and pHy RPEGFP, both carrying the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) ubiquitous promoter or the RPE65-specific promoter, respectively. For the two amplicon vectors, the GFP reporter gene was efficiently expressed in retinal pigment epithelial (RPE) cells but not in the adjacent photoreceptors. GFP expression was maximum as early as 2 days post-administration but decreased over time to become almost undetectable at 6 weeks postinjection. Super-transduction with a second amplicon vector, pHSVlac, reactivated expression of GFP in approximately 10% of the cells initially transduced at 2 days postinjection of pHy-CMVGFP or pHy-RPEGFP. Reactivation of transgene expression was transient, no GFP signal was detected 8 days after pHSVlac injection. In conclusion, HSV-1 amplicon vectors allow rapid and efficient, but transient, gene transfer in RPE cells following subretinal injection. PMID- 15889135 TI - Mechanism of efficient transfection of the nasal airway epithelium by hypotonic shock. AB - The main barrier to gene transfer in the airway epithelium is the low rate of apical endocytosis limiting naked DNA uptake. Deionized water is known to stimulate the exocytosis of numerous intracellular vesicles during hypotonic cell swelling, in order to expand plasma membrane and prevent cell lysis. This is followed by the phase of regulatory volume decrease (RVD), during which the excess plasma membrane is retrieved by intensive endocytosis. Here we show that the more hypotonic the DNA solution, the higher the transfection of the nasal tissue. P2 receptors are known to be involved in RVD and we demonstrate that some P2 agonists and a P2 antagonist impair transfection in a time-dependent manner. Our study strongly suggests that the nasal airway epithelial cells take up plasmid DNA in deionized water during RVD, within approximately half an hour. Our simple gene delivery system may constitute a promising method for respiratory tract gene therapy. PMID- 15889136 TI - Herpesvirus saimiri-based vector biodistribution using noninvasive optical imaging. AB - Herpesvirus saimiri (HVS) is capable of infecting a range of human cell types with high efficiency and the viral genome persists as high copy number, circular, nonintegrated episomes which segregate to progeny upon cell division. This allows the HVS-based vector to stably transduce a dividing cell population and provide sustained transgene expression for an extended period of time both in vitro and in vivo. Here we assess the dissemination of HVS-based vectors in vivo following intravenous and intraperitoneal administration. Bioluminescence imaging of an HVS based vector expressing luciferase demonstrates that the virus can infect and establish a persistent latent infection in a variety of mouse tissues. Moreover, the long-term in vivo maintenance of the HVS genome as a nonintegrated circular episome provided sustained expression of luciferase over a 10-week period. A particularly high level of transgene expression in the liver and the ability of HVS to infect and persist in hepatic stellate cells suggest that HVS-based vectors may have potential for the treatment of inherited and acquired liver diseases. PMID- 15889137 TI - Major role of local immune responses in antibody formation to factor IX in AAV gene transfer. AB - The risk of an immune response to the coagulation factor IX (F.IX) transgene product is a concern in gene therapy for the X-linked bleeding disorder hemophilia B. In order to investigate the mechanism of F.IX-specific lymphocyte activation in the context of adeno-associated viral (AAV) gene transfer to skeletal muscle, we injected AAV-2 vector expressing human F.IX (hF.IX) into outbred immune-competent mice. Systemic hF.IX levels were transiently detected in the circulation, but diminished concomitant with activation of CD4+ T and B cells. ELISPOT assays documented robust responses to hF.IX in the draining lymph nodes of injected muscle by day 14. Formation of inhibitory antibodies to hF.IX was observed over a wide range of vector doses, with increased doses causing stronger immune responses. A prolonged inflammatory reaction in muscle started at 1.5-2 months, but ultimately failed to eliminate transgene expression. By 1.5 months, hF.IX antigen re-emerged in circulation in approximately 70% of animals injected with high vector dose. Hepatic gene transfer elicited only infrequent and weaker immune responses, with higher vector doses causing a reduction in T cell responses to hF.IX. In summary, the data document substantial influence of target tissue, local antigen presentation, and antigen levels on lymphocyte responses to F.IX. PMID- 15889138 TI - Glycoprotein hormone receptors: link between receptor homodimerization and negative cooperativity. AB - The monomeric model of rhodopsin-like G protein-coupled receptors (GPCRs) has progressively yielded the floor to the concept of GPCRs being oligo(di)mers, but the functional correlates of dimerization remain unclear. In this report, dimers of glycoprotein hormone receptors were demonstrated in living cells, with a combination of biophysical (bioluminescence resonance energy transfer and homogenous time resolved fluorescence/fluorescence resonance energy transfer), functional and biochemical approaches. Thyrotropin (TSHr) and lutropin (LH/CGr) receptors form homo- and heterodimers, via interactions involving primarily their heptahelical domains. The large hormone-binding ectodomains were dispensable for dimerization but modulated protomer interaction. Dimerization was not affected by agonist binding. Observed functional complementation indicates that TSHr dimers may function as a single functional unit. Finally, heterologous binding competition studies, performed with heterodimers between TSHr and LH/CG-TSHr chimeras, demonstrated the unsuspected existence of strong negative cooperativity of hormone binding. Tracer desorption experiments indicated an allosteric behavior in TSHr and, to a lesser extent, in LH/CGr and FSHr homodimers. This study is the first report of homodimerization associated with negative cooperativity in rhodopsin-like GPCRs. As such, it may warrant revisitation of allosterism in the whole GPCR family. PMID- 15889139 TI - RMI1/NCE4, a suppressor of genome instability, encodes a member of the RecQ helicase/Topo III complex. AB - SGS1 encodes a DNA helicase whose homologues in human cells include the BLM, WRN, and RECQ4 genes, mutations in which lead to cancer-predisposition syndromes. Clustering of synthetic genetic interactions identified by large-scale genetic network analysis revealed that the genetic interaction profile of the gene RMI1 (RecQ-mediated genome instability, also known as NCE4 and YPL024W) was highly similar to that of SGS1 and TOP3, suggesting a functional relationship between Rmi1 and the Sgs1/Top3 complex. We show that Rmi1 physically interacts with Sgs1 and Top3 and is a third member of this complex. Cells lacking RMI1 activate the Rad53 checkpoint kinase, undergo a mitotic delay, and display increased relocalization of the recombination repair protein Rad52, indicating the presence of spontaneous DNA damage. Consistent with a role for RMI1 in maintaining genome integrity, rmi1Delta cells exhibit increased recombination frequency and increased frequency of gross chromosomal rearrangements. In addition, rmi1Delta strains fail to fully activate Rad53 upon exposure to DNA-damaging agents, suggesting that Rmi1 is also an important part of the Rad53-dependent DNA damage response. PMID- 15889140 TI - Oncogenic transcription factor Evi1 regulates hematopoietic stem cell proliferation through GATA-2 expression. AB - The ecotropic viral integration site-1 (Evi1) is an oncogenic transcription factor in murine and human myeloid leukemia. We herein show that Evi1 is predominantly expressed in hematopoietic stem cells (HSCs) in embryos and adult bone marrows, suggesting a physiological role of Evi1 in HSCs. We therefore investigate the role and authentic target genes of Evi1 in hematopoiesis using Evi1-/- mice, which die at embryonic day 10.5. HSCs in Evi1-/- embryos are markedly decreased in numbers in vivo with defective self-renewing proliferation and repopulating capacity. Notably, expression rate of GATA-2 mRNA, which is essential for proliferation of definitive HSCs, is profoundly reduced in HSCs of Evi1-/- embryos. Restoration of the Evi1 or GATA-2 expression in Evi1-/- HSCs could prevent the failure of in vitro maintenance and proliferation of HSC through upregulation of GATA-2 expression. An analysis of the GATA-2 promoter region revealed that Evi1 directly binds to GATA-2 promoter as an enhancer. Our results reveal that GATA-2 is presumably one of critical targets for Evi1 and that transcription factors regulate the HSC pool hierarchically. PMID- 15889141 TI - Intronic CA-repeat and CA-rich elements: a new class of regulators of mammalian alternative splicing. AB - We have recently identified an intronic polymorphic CA-repeat region in the human endothelial nitric oxide synthase (eNOS) gene as an important determinant of the splicing efficiency, requiring specific binding of hnRNP L. Here, we analyzed the position requirements of this CA-repeat element, which revealed its potential role in alternative splicing. In addition, we defined the RNA binding specificity of hnRNP L by SELEX: not only regular CA repeats are recognized with high affinity but also certain CA-rich clusters. Therefore, we have systematically searched the human genome databases for CA-repeat and CA-rich elements associated with alternative 5' splice sites (5'ss), followed by minigene transfection assays. Surprisingly, in several specific human genes that we tested, intronic CA RNA elements could function either as splicing enhancers or silencers, depending on their proximity to the alternative 5'ss. HnRNP L was detected specifically bound to these diverse CA elements. These data demonstrated that intronic CA sequences constitute novel and widespread regulatory elements of alternative splicing. PMID- 15889142 TI - Formation of a novel surface structure encoded by Salmonella Pathogenicity Island 2. AB - The type III secretion system (T3SS) encoded by Salmonella Pathogenicity Island 2 (SPI2) is essential for virulence and intracellular proliferation of Salmonella enterica. We have previously identified SPI2-encoded proteins that are secreted and function as a translocon for the injection of effector proteins. Here, we describe the formation of a novel SPI2-dependent appendage structure in vitro as well as on the surface of bacteria that reside inside a vacuole of infected host cells. In contrast to the T3SS of other pathogens, the translocon encoded by SPI2 is only present singly or in few copies at one pole of the bacterial cell. Under in vitro conditions, appendages are composed of a filamentous needle-like structure with a diameter of 10 nm that was sheathed with secreted protein. The formation of the appendage in vitro is dependent on acidic media conditions. We analyzed SPI2-encoded appendages in infected cells and observed that acidic vacuolar pH was not required for induction of SPI2 gene expression, but was essential for the assembly of these structures and their function as translocon for delivery of effector proteins. PMID- 15889143 TI - Beta1 integrins regulate mammary gland proliferation and maintain the integrity of mammary alveoli. AB - Integrin-extracellular matrix interactions play important roles in the coordinated integration of external and internal cues that are essential for proper development. To study the role of beta1 integrin in the mammary gland, Itgbeta1(flox/flox) mice were crossed with WAPiCre transgenic mice, which led to specific ablation of beta1 integrin in luminal alveolar epithelial cells. In the beta1 integrin mutant mammary gland, individual alveoli were disorganized resulting from alterations in cell-basement membrane associations. Activity of focal adhesion kinase (FAK) was also decreased in mutant mammary glands. Luminal cell proliferation was strongly inhibited in beta1 integrin mutant glands, which correlated with a specific increase of p21 Cip1 expression. In a p21 Cip1 null background, there was a partial rescue of BrdU incorporation, providing in vivo evidence linking p21 Cip1 to the proliferative defect observed in beta1 integrin mutant glands. A connection between p21 Cip1 and beta1 integrin as well as FAK was also established in primary mammary cells. These results point to the essential role of beta1 integrin signaling in mammary epithelial cell proliferation. PMID- 15889144 TI - Phosducin-like protein acts as a molecular chaperone for G protein betagamma dimer assembly. AB - Phosducin-like protein (PhLP) is a widely expressed binding partner of the G protein betagamma subunit dimer (Gbetagamma). However, its physiological role is poorly understood. To investigate PhLP function, its cellular expression was blocked using RNA interference, resulting in inhibition of Gbetagamma expression and G protein signaling. This inhibition was caused by an inability of nascent Gbetagamma to form dimers. Phosphorylation of PhLP at serines 18-20 by protein kinase CK2 was required for Gbetagamma formation, while a high-affinity interaction of PhLP with the cytosolic chaperonin complex appeared unnecessary. PhLP bound nascent Gbeta in the absence of Ggamma, and S18-20 phosphorylation was required for Ggamma to associate with the PhLP-Gbeta complex. Once Ggamma bound, PhLP was released. These results suggest a mechanism for Gbetagamma assembly in which PhLP stabilizes the nascent Gbeta polypeptide until Ggamma can associate, resulting in membrane binding of Gbetagamma and release of PhLP to catalyze another round of assembly. PMID- 15889145 TI - The yeast lipin Smp2 couples phospholipid biosynthesis to nuclear membrane growth. AB - Remodelling of the nuclear membrane is essential for the dynamic changes of nuclear architecture at different stages of the cell cycle and during cell differentiation. The molecular mechanism underlying the regulation of nuclear membrane biogenesis is not known. Here we show that Smp2, the yeast homologue of mammalian lipin, is a key regulator of nuclear membrane growth during the cell cycle. Smp2 is phosphorylated by Cdc28/Cdk1 and dephosphorylated by a nuclear/endoplasmic reticulum (ER) membrane-localized CPD phosphatase complex consisting of Nem1 and Spo7. Loss of either SMP2 or its dephosphorylated form causes transcriptional upregulation of key enzymes involved in lipid biosynthesis concurrent with a massive expansion of the nucleus. Conversely, constitutive dephosphorylation of Smp2 inhibits cell division. We show that Smp2 associates with the promoters of phospholipid biosynthetic enzymes in a Nem1-Spo7-dependent manner. Our data suggest that Smp2 is a critical factor in coordinating phospholipid biosynthesis at the nuclear/ER membrane with nuclear growth during the cell cycle. PMID- 15889146 TI - Replication fork blockage by RTS1 at an ectopic site promotes recombination in fission yeast. AB - Homologous recombination is believed to play important roles in processing stalled/blocked replication forks in eukaryotes. In accordance with this, recombination is induced by replication fork barriers (RFBs) within the rDNA locus. However, the rDNA locus is a specialised region of the genome, and therefore the action of recombinases at its RFBs may be atypical. We show here for the first time that direct repeat recombination, dependent on Rad22 and Rhp51, is induced by replication fork blockage at a site-specific RFB (RTS1) within a 'typical' genomic locus in fission yeast. Importantly, when the RFB is positioned between the direct repeat, conservative gene conversion events predominate over deletion events. This is consistent with recombination occurring without breakage of the blocked fork. In the absence of the RecQ family DNA helicase Rqh1, deletion events increase dramatically, which correlates with the detection of one-sided DNA double-strand breaks at or near RTS1. These data indicate that Rqh1 acts to prevent blocked replication forks from collapsing and thereby inducing deletion events. PMID- 15889147 TI - Urokinase-induced signaling in human vascular smooth muscle cells is mediated by PDGFR-beta. AB - Urokinase (uPA)-induced signaling in human vascular smooth muscle cells (VSMC) elicits important cellular functional responses, such as cell migration and proliferation. However, how intracellular signaling is linked to glycolipid anchored uPA receptor (uPAR) is unknown. We provide evidence that uPAR activation by uPA induces its association with platelet-derived growth factor receptor (PDGFR)-beta. The interaction results in PDGF-independent PDGFR-beta activation by phosphorylation of cytoplasmic tyrosine kinase domains and receptor dimerization. Association of the receptors as well as the tyrosine kinase activity of PDGFR-beta are decisive in mediating uPA-induced downstream signaling that regulates VSMC migration and proliferation. These findings provide a molecular basis for mechanisms VSMC use to induce uPAR- and PDGFR-directed signaling. The processes may be relevant to VSMC function and vascular remodeling. PMID- 15889148 TI - Sensing wetness: a new role for the bacterial flagellum. AB - We have uncovered a new role for the bacterial flagellum in sensing external wetness. An investigation into why mutants in the chemotaxis signaling pathway of Salmonella typhimurium exhibit fewer and shorter flagella than wild-type when propagated on a surface, first showed that the mutants downregulate only a small set of genes on swarm media--class 3 or 'late' motility genes, and genes associated with the pathogenicity island SPI-1 TTSS (type three secretion system). Based on observations that swarm colonies of the mutants appear less hydrated, we tested a model in which the flagellum itself is a sensor: suboptimal external hydration interferes with secretion of flagellin subunits, inhibiting filament growth and blocking normal export of the class 3 transcription inhibitor FlgM. We provide strong experimental support for the model. In addition, the data show that the flagellar and SPI-1 TTSS are coupled via regulatory proteins. These studies implicate the flagellum, a bacterial organ for motility, in sensing the external environment to modulate not only its own biogenesis but other physiological functions as well. PMID- 15889149 TI - Deficiency of LKB1 in skeletal muscle prevents AMPK activation and glucose uptake during contraction. AB - Recent studies indicate that the LKB1 tumour suppressor protein kinase is the major "upstream" activator of the energy sensor AMP-activated protein kinase (AMPK). We have used mice in which LKB1 is expressed at only approximately 10% of the normal levels in muscle and most other tissues, or that lack LKB1 entirely in skeletal muscle. Muscle expressing only 10% of the normal level of LKB1 had significantly reduced phosphorylation and activation of AMPKalpha2. In LKB1 lacking muscle, the basal activity of the AMPKalpha2 isoform was greatly reduced and was not increased by the AMP-mimetic agent, 5-aminoimidazole-4-carboxamide riboside (AICAR), by the antidiabetic drug phenformin, or by muscle contraction. Moreover, phosphorylation of acetyl CoA carboxylase-2, a downstream target of AMPK, was profoundly reduced. Glucose uptake stimulated by AICAR or muscle contraction, but not by insulin, was inhibited in the absence of LKB1. Contraction increased the AMP:ATP ratio to a greater extent in LKB1-deficient muscles than in LKB1-expressing muscles. These studies establish the importance of LKB1 in regulating AMPK activity and cellular energy levels in response to contraction and phenformin. PMID- 15889150 TI - Prostaglandin D2 induces nuclear import of the sex-determining factor SOX9 via its cAMP-PKA phosphorylation. AB - During mammalian gonadal development, nuclear import/export of the transcription factor SOX9 is a critical step of the Sry-initiated testis-determining cascade. In this study, we identify a molecular mechanism contributing to the SOX9 nuclear translocation in NT2/D1 cells, which is mediated by the prostaglandin D2 (PGD2) signalling pathway via stimulation of its adenylcyclase-coupled DP1 receptor. We find that activation of cAMP-dependent protein kinase A (PKA) induces phosphorylation of SOX9 on its two S64 and S181 PKA sites, and its nuclear localization by enhancing SOX9 binding to the nucleocytoplasmic transport protein importin beta. Moreover, in embryonic gonads, we detect a male-specific prostaglandin D synthase expression and an active PGD2 signal at the time and place of SOX9 expression. We thus propose a new step in the sex-determining cascade where PGD2 acts as an autocrine factor inducing SOX9 nuclear translocation and subsequent Sertoli cell differentiation. PMID- 15889151 TI - Developmental specificity of auxin response by pairs of ARF and Aux/IAA transcriptional regulators. AB - The plant hormone auxin elicits many specific context-dependent developmental responses. Auxin promotes degradation of Aux/IAA proteins that prevent transcription factors of the auxin response factor (ARF) family from regulating auxin-responsive target genes. Aux/IAAs and ARFs are represented by large gene families in Arabidopsis. Here we show that stabilization of BDL/IAA12 or its sister protein IAA13 prevents MP/ARF5-dependent embryonic root formation whereas stabilized SHY2/IAA3 interferes with seedling growth. Although both bdl and shy2 2 proteins inhibited MP/ARF5-dependent reporter gene activation, shy2-2 was much less efficient than bdl to interfere with embryonic root initiation when expressed from the BDL promoter. Similarly, MP was much more efficient than ARF16 in this process. When expressed from the SHY2 promoter, both shy2-2 and bdl inhibited cell elongation and auxin-induced gene expression in the seedling hypocotyl. By contrast, gravitropism and auxin-induced gene expression in the root, which were promoted by functionally redundant NPH4/ARF7 and ARF19 proteins, were inhibited by shy2-2, but not by bdl protein. Our results suggest that auxin signals are converted into specific responses by matching pairs of coexpressed ARF and Aux/IAA proteins. PMID- 15889152 TI - Sec17p and HOPS, in distinct SNARE complexes, mediate SNARE complex disruption or assembly for fusion. AB - SNARE functions during membrane docking and fusion are regulated by Sec1/Munc18 (SM) chaperones and Rab/Ypt GTPase effectors. These functions for yeast vacuole fusion are combined in the six-subunit HOPS complex. HOPS facilitates Ypt7p nucleotide exchange, is a Ypt7p effector, and contains an SM protein. We have dissected the associations and requirements for HOPS, Ypt7p, and Sec17/18p during SNARE complex assembly. Vacuole SNARE complexes bind either Sec17p or the HOPS complex, but not both. Sec17p and its co-chaperone Sec18p disassemble SNARE complexes. Ypt7p regulates the reassembly of unpaired SNAREs with each other and with HOPS, forming HOPS.SNARE complexes prior to fusion. After HOPS.SNARE assembly, lipid rearrangements are still required for vacuole content mixing. Thus, Sec17p and HOPS have mutually exclusive interactions with vacuole SNAREs to mediate disruption of SNARE complexes or their assembly for docking and fusion. Sec17p may displace HOPS from SNAREs to permit subsequent rounds of fusion. PMID- 15889153 TI - H662 is the linchpin of ATP hydrolysis in the nucleotide-binding domain of the ABC transporter HlyB. AB - The ABC transporter HlyB is a central element of the HlyA secretion machinery, a paradigm of Type I secretion. Here, we describe the crystal structure of the HlyB NBD (nucleotide-binding domain) with H662 replaced by Ala in complex with ATP/Mg2+. The dimer shows a composite architecture, in which two intact ATP molecules are bound at the interface of the Walker A motif and the C-loop, provided by the two monomers. ATPase measurements confirm that H662 is essential for activity. Based on these data, we propose a model in which E631 and H662, highly conserved among ABC transporters, form a catalytic dyad. Here, H662 acts as a 'linchpin', holding together all required parts of a complicated network of interactions between ATP, water molecules, Mg2+, and amino acids both in cis and trans, necessary for intermonomer communication. Based on biochemical experiments, we discuss the hypothesis that substrate-assisted catalysis, rather than general base catalysis might operate in ABC-ATPases. PMID- 15889154 TI - Impaired maturation of myeloid progenitors in mice lacking novel Polycomb group protein MBT-1. AB - Polycomb group (PcG) proteins participate in DNA-binding complexes with gene repressing activity, many of which have been highlighted for their involvement in hematopoiesis. We have identified a putative PcG protein, termed MBT-1, that is associated with Rnf2, an in vivo interactor of PcG proteins. MBT-1 structurally resembles the H-L(3)MBT protein, whose deletion is predicted to be responsible for myeloid hematopoietic malignancies. The human MBT-1 gene is located on chromosome 6q23, a region frequently deleted in leukemia cells, and shows a transient expression spike in response to maturation-inducing stimuli in myeloid leukemia cells. MBT-1(-/-) myeloid progenitor cells exhibit a maturational deficiency but maintain normal proliferative activities. This results in the accumulation of immature myeloid progenitors and hence, a marked decrease of mature myeloid blood cells, causing the MBT-1(-/-) mice to die of anemia during a late embryonic stage. Together, we conclude that MBT-1 specifically regulates the maturational advancement of myeloid progenitor cells during transitions between two developmental stages. We also show that MBT-1 appears to influence myelopoiesis by transiently enhancing p57(KIP2) expression levels. PMID- 15889155 TI - Leukemia-derived dendritic cells in acute myeloid leukemia exhibit potent migratory capacity. PMID- 15889156 TI - Prognostic implication of FLT3 and Ras gene mutations in patients with acute promyelocytic leukemia (APL): a retrospective study from the European APL Group. AB - Internal tandem duplications (ITDs) of the FLT3 gene have been observed in about 35% of APL cases. If FLT3-ITD is associated with a worse outcome in patients with acute myeloid leukemia (AML) in general, its prognostic value in acute promyelocytic leukemia (APL) is still a matter of debate. We investigated incidence, associated clinical features, and prognostic implication of FLT3-ITD, but also FLT3-D835 point mutation and N-Ras or K-Ras mutations in 119 APL patients, all prospectively enrolled in the two consecutive APL-93 and APL-2000 trials. Mutation incidences were 38, 20, and 4%, for FLT3-ITD, FLT3-D835, and Ras, respectively. The presence of FLT3-ITD was associated with high white blood cell count, high Sanz index, M3-variant subtype, and V/S PML-RAR alpha isoforms. Complete remission (CR), induction death, and death in CR rates were not affected by FLT3 or Ras mutations, as well as cumulative incidence of relapse. However, a trend for a shorter overall survival (P=0.09) was observed in FLT3-ITD patients, because of a very poor postrelapse survival (P=0.02). This feature, which has been also reported in patients with AML in general, is suggestive of an underlying genetic instability in FLT3-ITD patients, leading to the acquisition of additional unknown bad-prognosis gene mutations at relapse. PMID- 15889157 TI - Transcriptional regulation of parathyroid hormone-related protein promoter P3 by ETS-1 in adult T-cell leukemia/lymphoma. AB - Parathyroid hormone-related protein (PTHrP) plays a primary role in the development of humoral hypercalcemia of malignancy seen in the majority of adult T-cell leukemia/lymphoma (ATLL) patients with human T-cell lymphotropic virus type-1 (HTLV-1) infection. HTLV-1 Tax has been shown to complex with ETS-1 and SP1 to transactivate the PTHrP P3 promoter. Previously, we established a SCID/bg mouse model of human ATL with RV-ATL cells and showed that PTHrP expression was independent of Tax. In this study, we report an inverse correlation of PTHrP with tax/rex mRNA in multiple HTLV-1-positive cell lines and RV-ATL cells. Stimulation of Jurkat T cells with PMA/ionomycin upregulated the PTHrP P3 promoter by a previously characterized Ets binding site and also induced protein/DNA complex formation identical to that observed in RV-ATL cells. Further, we provide evidence that cotransfection with Ets-1 and constitutively active Mek-1 in HTLV-1 negative transformed T cells with stimulation by PMA/ionomycin not only resulted in a robust induction of PTHrP P3 but also formed a complex with ETS-1/P3 EBS similar to that in ATLL cells. Our data demonstrate that transcriptional regulation of PTHrP in ATLL cells can be controlled by T-cell receptor signaling and the ETS and MAPK ERK pathway in a Tax-independent manner. PMID- 15889158 TI - Lonafarnib reduces the resistance of primitive quiescent CML cells to imatinib mesylate in vitro. AB - Recent studies indicate that a rare population of primitive quiescent BCR-ABL(+) cells are innately insensitive to imatinib mesylate (IM) and persist after IM therapy of patients with chronic myeloid leukemia (CML). New approaches to the eradication of these cells are therefore likely to be crucial to the development of curative therapies for CML. We have now found that Ara-C, LY294002 (a PI-3 (phosphatidylinositol-3' kinase) kinase inhibitor), 17AAG (a heat-shock protein (HSP)-90 antagonist) and lonafarnib (a farnesyltransfease inhibitor) all enhance the toxicity of IM on K562 cells and on the total CD34(+) leukemic cell population from chronic phase CML patients. However, for quiescent CD34(+) leukemic cells, this was achieved only by concomitant exposure of the cells to lonafarnib. Ara-C or LY294002 alone blocked the proliferation of these cells but did not kill them, and Ara-C, LY294002 or 17AAG in combination with IM enhanced the cytostatic effect of IM but did not prevent the subsequent regrowth of the surviving leukemic cells. These studies demonstrate the importance of in vitro testing of novel agents on the subset of primary leukemic cells most likely to determine long-term treatment outcomes in vivo. PMID- 15889159 TI - Discrete serum protein signatures discriminate between human retrovirus associated hematologic and neurologic disease. AB - The human T-cell leukemia virus type I (HTLV-I) is the causative agent for adult T-cell leukemia (ATL) and HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Approximately 5% of infected individuals will develop either disease and currently there are no diagnostic tools for early detection or accurate assessment of disease state. We have employed high-throughput expression profiling of serum proteins using mass spectrometry to identify protein expression patterns that can discern between disease states of HTLV-I-infected individuals. Our study group consisted of 42 ATL, 50 HAM/TSP, and 38 normal controls. Spectral peaks corresponding to peptide ions were generated from MS-TOF data. We applied Classification and Regression Tree analysis to build a decision algorithm, which achieved 77% correct classification rate across the three groups. A second cohort of 10 ATL, 10 HAM and 10 control samples was used to validate this result. Linear discriminate analysis was performed to verify and visualize class separation. Affinity and sizing chromatography coupled with tandem mass spectrometry was used to identify three peaks specifically overexpressed in ATL: an 11.7 kDa fragment of alpha trypsin inhibitor, and two contiguous fragments (19.9 and 11.9 kDa) of haproglobin-2. To the best of our knowledge, this is the first application of protein profiling to distinguish between two disease states resulting from a single infectious agent. PMID- 15889160 TI - Catalytic asymmetric hydroamination of non-activated olefins. AB - Hydroamination is a highly atom-economical process in which an amine N-H functionality is added to an unsaturated carbon-carbon linkage. This potentially highly useful process gives access to various nitrogen-containing basic and fine chemicals as well as naturally occurring alkaloid skeletons. Asymmetric hydroamination reactions promoted by chiral catalysts are particularly attractive. This review highlights recent progress in the development of early transition metal catalysts for the asymmetric hydroamination of non-activated alkenes. PMID- 15889161 TI - Inhibition studies on salicylate synthase. AB - Analogues of chorismate and isochorismate were designed and tested as potential inhibitors in the first inhibition study against a salicylate synthase. PMID- 15889162 TI - Facile synthesis of multisubstituted buta-1,3-dienes via Suzuki-Miyaura and Kumada cross-coupling strategy of 2,4-diiodo-buta-1-enes with arylboronic acids and Grignard reagents. AB - One-pot Suzuki-Miyaura-type and Kumada-type cross-coupling reactions of 2,4 diiodo-buta-1-enes with arylboronic acids and alkyl/aryl magnesium bromides were carried out in the presence of accessibly simple catalysts under mild conditions. As a result, some 1,1,2-trisubstituted buta-1,3-dienes were obtained including the Tamoxifen-type, which have potential adjuvant therapy in women who have suffered from breast cancer and cyclooxygenase-2-type (COX-2-type) inhibitors, some of which have been proved to elicit efficient anti-inflammatory analgesic activities and less adverse gastrointestinal side effects and to be very useful in the prophylactic treatment of a wide variety of cancers and neurodegenerative disorders. PMID- 15889163 TI - Enhancement of the chemical semantic web through the use of InChI identifiers. AB - Molecules, as defined by connectivity specified via the International Chemical Identifier (InChI), are precisely indexed by major web search engines so that Internet tools can be transparently used for unique structure searches. PMID- 15889164 TI - Efficient solid phase synthesis of benzo[1,2,3]thiadiazoles and related structures. AB - The first solid-phase synthesis of benzo[1,2,3]thiadiazoles was achieved by starting from resin bound ortho bromo or iodo triazenes and using a functionalization on cleavage. PMID- 15889165 TI - Protonated nitro group: structure, energy and conjugation. AB - Structure of protonated nitro compounds was investigated by calculations at the levels MP2(FC)/6-311++G(2d,2p)//MP2(FC)/6-311++G(2d,2p)(nitromethane and reference compounds) or B3LYP/6-311+G(d,p)//B3LYP/6-311+G(d,p)(nitrobenzene and its 18 meta- and para-substituted derivatives). The group NO2H+ reveals many similarities with the isoelectronic group CO2H as the preferred conformation, conformational equilibrium, and stabilization by interaction (resonance) within the group quantified by means of isodesmic reactions. However, there is a difference in the interaction with donor groups (for instance in 4-nitroaniline) that is much stronger with NO2H+ than with CO2H. This interaction may be called resonance and may be described by standard resonance formulas, but these formulas predict only partially the geometry and cannot explain the great interaction energy. PMID- 15889166 TI - New hydroxy-pyrazoline intermediates, subtle regio-selectivity and relative reaction rate variations observed during acid catalyzed and neutral pyrazole cyclization. AB - Arylhydrazines 4 and 1,3-dicarbonyl compounds 5 react to form pyrazoles by loss of water via hydrazone isomer pairs 6 and 7 which give rise to two possible regio isomers. Occasionally, 3-hydroxy-3,4-dihydropyrazoles or hydroxy-pyrazolines 9 and 9 are observed as stable isolatable intermediates that can be fully characterized prior to loss of the second molecule of water that gives rise to pyrazoles 10 and 11. Fully characterized examples of intermediates of type 8 and 9 are relatively rare. We studied the reaction series where R = CH3, CHF2 and CF3 and Ar = Ph and 5-methanesulfonylpyridin-2-yl, (Scheme 2), and observed differences in properties between kinetic behavior and regio-isomerism depending on the degree of electron-withdrawing capability of the R and Ar substituents. The reaction conditions that caused cyclization to pyrazoles varied from direct condensation of the hydrazine and 1,3-dicarbonyl compounds, to reactions requiring catalytic quantities of sulfuric acid to sulfuric acid in excess. Unexpected regio-selectivity was observed in the case of R = CF3 that depended upon the reaction conditions. PMID- 15889167 TI - Prebiotic carbohydrate synthesis: zinc-proline catalyzes direct aqueous aldol reactions of alpha-hydroxy aldehydes and ketones. AB - Zn-proline catalyzed aldolisation of glycoladehyde gave mainly tetroses whereas in the cross-aldolisation of glycoladehyde and rac-glyceraldehyde, pentoses accounted for 60% of the sugars formed with 20% of ribose. PMID- 15889168 TI - Syntheses and copper(II)-dependent DNA photocleavage by acridine and anthracene 1,10-phenanthroline conjugate systems. AB - We report the syntheses and characterization of a series of compounds based on 1,10-phenanthroline covalently tethered, at the 2 and 9 positions, to either two benzene, naphthalene, acridine or anthracene chromophores. The acridine and anthracene derivatives are shown to efficiently cleave pUC19 plasmid DNA upon irradiation with ultraviolet light (pH = 7.0, 22 degrees C, 350 nm). Furthermore, photocleavage levels are markedly increased by the addition of Cu2+ to the DNA photolysis reactions. Interestingly, when the concentrations of the anthracene compounds are lowered from 35 microM to 0.25 microM, the reverse trend is observed. DNA photocleavage is markedly reduced in the presence of copper(II). PMID- 15889169 TI - Mutual induced fit in cyclodextrin-rocuronium complexes. AB - The binding of rocuronium bromide to 6-perdeoxy-6-per(4-carboxyphenyl)thio-gamma cyclodextrin sodium salt, displays biphasic behaviour characteristic of the formation of a binary and 2 : 1 ternary guest-host complex in aqueous solution. Thermodynamic and structural data on this sequential complexation process can be rationalised within a single model involving switching of the conformational equilibria of both the rocuronium bromide and cyclodextrin molecules. Isothermal titration calorimetry (ITC), NMR and fluorescence experiments in solution, together with X-ray crystallography and molecular modelling, suggest that in order to induce encapsulation both rocuronium bromide and the modified cyclodextrin undergo conformational changes. Ring A of rocuronium bromide 'switches' from the more sterically encumbered chair to the sterically less demanding twist-boat, whilst the modified cyclodextrin "opens" its cavity to allow the steroid to enter. The recognition and mutual induced fit between cyclodextrin and steroid represents a classic example of dynamic host-guest chemistry. PMID- 15889170 TI - The glucosinolate-myrosinase system. New insights into enzyme-substrate interactions by use of simplified inhibitors. AB - Myrosinase, a thioglucoside glucohydrolase, is the only enzyme able to hydrolyse glucosinolates, a unique family of molecules bearing an anomeric O-sulfated thiohydroximate function. Non-hydrolysable myrosinase inhibitors have been devised and studied for their biological interaction. Diverse modifications of the O-sulfate moiety did not result in a significant inhibitory effect, whereas replacing the D-glucopyrano residue by its carba-analogue allowed inhibition to take place. X-Ray experiments carried out after soaking allowed for the first time inclusion of a non-hydrolysable inhibitor inside the enzymatic pocket. Structural tuning of the aglycon part in its pocket is being used as a guide for the development of simplified and more potent inhibitors. PMID- 15889171 TI - An extensive study of bromination of cis,trans,trans-1,5,9-cyclododecatriene: product structures and conformations. AB - Bromine has been added to cis,trans,trans-1,5,9-cyclododecatriene under various reaction conditions. All expected direct addition products have been isolated, and their structures have been determined by microanalysis, NMR and X-ray crystallography. Advanced NMR techniques were used to determine solution conformations of several of the compounds, enabling comparison with the solid state conformations obtained by crystallography. PMID- 15889172 TI - Deprotonation-electrophile trapping of terminal epoxides. AB - Organolithium-induced deprotonation of terminal epoxides in the presence of appropriate diamine ligands allows trapping with a range of electrophiles, yielding functionalised di- and tri-substituted epoxides in good yields and with control of stereochemistry at the epoxide. PMID- 15889173 TI - Bioreduction activated prodrugs of camptothecin: molecular design, synthesis, activation mechanism and hypoxia selective cytotoxicity. AB - Several water-soluble derivatives (CPT3, CPT3a-d) of camptothecin (CPT) were synthesized, among which CPT3 bearing an N,N'-dimethyl-1-aminoethylcarbamate side chain was further conjugated with reductively eliminating structural units of indolequinone, 4-nitrobenzyl alcohol and 4-nitrofuryl alcohol to produce novel prodrugs of camptothecin (CPT4-6). All CPT derivatives were of lower cytotoxicity than their parent compound of CPT. In contrast, CPT4 and CPT6 showed higher hypoxia selectivity of cytotoxicity towards tumor cells than CPT. A mechanism by which a representative prodrug CPT4 is activated in the presence of DT-diaphorase to release CPT was also discussed. The bioreduction activated CPT prodrugs including CPT4 and CPT6 are identified to be promising for application to the hypoxia targeting tumor chemotherapy. PMID- 15889174 TI - Tuning the size of macrocyclic cavities in trianglimine macrocycles. AB - The synthesis of aromatic dicarboxaldehydes is described along with their reactivity in the [3 + 3] cyclocondensation reaction with (1R,2R) diaminocyclohexane to give trianglimine macrocycles. In particular, the scope and limitation of the reaction with regard to complete control of the cavity size of the macrocycles is discussed producing a total of 11 macrocycles with different cavity sizes ranging from 9 to 23 angstroms. PMID- 15889175 TI - Synthesis of a phenyl thio-beta-D-galactopyranoside library from 1,5-difluoro-2,4 dinitrobenzene: discovery of efficient and selective monosaccharide inhibitors of galectin-7. AB - The galectins are a family of [small beta]-galactoside-binding proteins that have been implicated in cancer and inflammation processes. Herein, we report the synthesis of a library of 28 compounds that was tested for binding to galectins 1, -3, -7, -8N and -9N. An aromatic nucleophilic substitution reaction between 1,5-difluoro-2,4-dinitrobenzene and a galacto thiol gave 5-fluoro-2,4 dinitrophenyl 2,3,4,6-tetra-O-acetyl-1-thio-beta-D-galactopyranoside. This versatile intermediate was then modified in a two dimensional manner: either by further substitution of the second fluoride by amines or thiols, or by reduction of the nitro groups and acylation of the resulting amines, or both. Deacetylation then gave a library of aromatic beta-galactosides that showed variable inhibitory activity against the different galectins, as shown by screening with a fluorescence-polarisation assay. Particularly efficient inhibitors were found against galectin-7, while less impressive enhancements of inhibitor affinity over methyl beta-D-galactopyranoside were found for galectin-1, -3, -8N and -9N. The best inhibitors against galectin-7 showed significantly higher affinity (K(d) as low as 140 microM) than both beta-methyl galactoside (K(d) 4.8 mM) and the unsubstituted beta-phenyl thiogalactoside (non-inhibitory). The best inhibitors against galectin-7 were poor against the other galectins and thus have potential as structurally simple and selective tools for dissecting biological functions of galectin-7. PMID- 15889176 TI - Sc(OTf)3-catalyzed efficient synthesis of beta,beta-bis(indolyl) ketones by the double indolylation of acetic acid 2-methylene-3-oxobutyl ester. AB - In the presence of 5 mol% Sc(OTf)3, double indolylation of acetic acid 2 methylene-3-oxo-butyl ester with differently substituted indoles readily afforded beta,beta-bis(indolyl) ketones. The reaction may proceed via a Sc(OTf)3-catalyzed SN2'-type substitution and subsequent conjugate addition. PMID- 15889177 TI - Model compounds for (6-4) photolyases: a comparative flavin induced cleavage study of oxetanes and thietanes. AB - Thietanes were used in the past as mimics for an unstable oxetane intermediate formed during the repair of mutagenic (6-4) lesions. The thietane derivatives were found to be not repaired, raising the question of how well thietanes are cleaved by single electron donation compared to oxetanes. We have prepared two flavin-containing oxetane and thietane model compounds for the (6-4) photolyase catalyzed repair process and we show that both are efficiently cleaved by a reduced and deprotonated flavin. Thietanes are therefore excellent models. The lack of their repair can be attributed to lack of binding. PMID- 15889178 TI - The synthesis of water soluble decalin-based thiols and S-nitrosothiols--model systems for studying the reactions of nitric oxide with protein thiols. AB - The syntheses of three decalin-based tert-thiols displaying varying degrees of solubility in aqueous milieu are described. The S-nitroso derivatives of these compounds have also been prepared and the structures of two of these determined by single crystal X-ray diffraction. These compounds have been designed for studying the interaction of nitric oxide (NO) with thiols under physiological conditions. PMID- 15889179 TI - Chemoenzymatic synthesis of carbasugars from iodobenzene. AB - The versatile enantiopure cis-dihydrodiol metabolite 1, formed by bacterial metabolism of iodobenzene, has been used for the synthesis of the pyranose carbasugars (pseudosugars) carba-beta-D-altropyranose 2, carba-alpha-L galactopyranose 3, carba-beta-D-idopyranose 4 and carba-beta-L-glucopyranose 5. Substitution of the iodine atom by a carbomethoxy group, stereoselective catalytic hydrogenation of an alpha,beta-unsaturated ester, and regioselective inversion of one or two allylic chiral centres are the key steps used in the synthesis of carbasugars 2-5. The relative and absolute configurations of compounds 2-5 were established by a combination of stereochemical correlation, X ray crystallography and 1H-NMR spectroscopy. PMID- 15889180 TI - Cyclization reactions of 1-[3'-hydroxy-2'-(hydroxymethyl)prop-1'-enyl]pyrimidine nucleobases: intramolecular Michael additions to the C(5)=C(6) bonds and intramolecular dehydrations. AB - The tendency of a series of acyclic nucleoside analogues 1a-f to undergo intramolecular cyclization reactions was investigated. All compounds, when treated with NaOD, were in equilibrium with the bicyclic compounds 2a-f, arising from Michael addition of a hydroxy group to the C(5)=C(6) bonds. Derivatives of 2,4-pyrimidinediones (1a,b) had the highest tendency to undergo intramolecular Michael addition when treated with triethylamine, whereas the cyclization of 4 amino-2-pyridones (1c-f) proceeded best with acid. The exocyclic double bond of was essential for the cyclization to occur. Commonly used N-protecting groups as the benzoyl- and the dibutylaminomethylene group enhanced cyclization. Under acidic anhydrous conditions 1b and 1e cyclized to the 2,4'-anhydro compounds 1b and 1e. PMID- 15889181 TI - Absolute configuration and predominant conformations of 1,1-dimethyl-2 phenylethyl phenyl sulfoxide. AB - The absolute configuration of the (+)-1,1-dimethyl-2-phenylethyl phenyl sulfoxide is determined to be (R), using three different chiroptical spectroscopic methods, namely vibrational circular dichroism (VCD), electronic circular dichroism (ECD) and specific rotation. Four solution conformations are identified for 1,1 dimethyl-2-phenylethyl phenyl sulfoxide. In each of the methods used, experimental data for the enantiomers of 1,1-dimethyl-2-phenylethyl phenyl sulfoxide were measured in the solution phase and concomitant quantum mechanical calculations of corresponding properties were carried out using density functional theory with B3LYP functional and 6-31G* and 6-31+G basis sets. Additional VCD and ECD calculations were also undertaken with 6-311G(2d,2p) basis set. A comparison of theoretically predicted data with the corresponding experimental data has allowed us to elucidate the absolute configuration and predominant conformations of (+)-1,1-dimethyl-2-phenylethyl phenyl sulfoxide. PMID- 15889182 TI - Rapid, iterative assembly of octyl alpha-1,6-oligomannosides and their 6-deoxy equivalents. AB - Mycobacterium tuberculosis is the cause of the deadly human disease tuberculosis. In studies over the last 40 years it has been revealed that this organism possesses a complex cell wall including glycophospholipids such as the phosphatidylinositiol mannosides (PIMs), lipomannan (LM) and lipoarabinomannan (LAM). These glycolipids all contain a common alpha-1,6-linked mannoside core, and the higher PIMs and LAM possess alpha-1,2-linked mannosyl residues. It has been shown that simple alpha-1,6-linked oligomannosides can act as substrates for alpha-1,6-mannosyltransferases in mycobacteria. Here we report a simple iterative synthesis of a series of hydrophobic octyl alpha-1,6-linked oligomannosides from mono- through to tetrasaccharides. We have utilized a single thioglycoside donor and alcohol acceptor. Further, we have developed conditions for the conversion of each of these compounds to the 6-deoxy congeners. Deoxygenation of the 6-position of the terminal mannosyl residue should prevent these compounds acting as substrates for the abundant alpha-1,6-mannosyltransferases in mycobacteria and should permit detection of the elusive alpha-1,2-mannosyltransferase activity responsible for elaboration of LM to mature LAM and the biosynthesis of the higher PIMs. PMID- 15889183 TI - Thiacalix[4]arene derivatives as radium ionophores: a study on the requirements for Ra2+ extraction. AB - The synthesis and NOE-based structural characterization is described of thiacalix[4]arene tricarboxylic acid (7), thiacalix[4]crown-5 and -6 monocarboxylic acids (2 and 5), and the bis(N-methylsulfonyl)thiacalix[4]crowns-5 and -6 (4a,b). The 226Ra2+ selectivity coefficients, log(K(Ra)ex/K(M)ex), of the new thiacalix[4]arene derivatives are compared directly with those of thiacalix[4]crown-5 and -6 (1a,b), thiacalix[4]crown-5 and -6 dicarboxylic acids (3a,b), and thiacalix[4]arene di- and tetracarboxylic acids (6 and 8). Thiacalix[4]arene dicarboxylic acid (6) already exhibits a high 226Ra2+ selectivity, but this is significantly improved in the case of 3b, having an additional crown-(6-)ether bridge. The covalent combination of a crown ether and carboxylic acid substituents as in the thiacalix[4]arenes 2,3a,b,4a,b, and 5 gives a better 226Ra2+ selectivity in the presence of Sr2+ or Ba2+ than mixtures of dibenzo-21-crown-7 and thiacalix[4]arene dicarboxylic acid (6) or of pentadecanoic acid and thiacalix[4]crown-6 (1b). PMID- 15889184 TI - Detoxification pathways of the phytoalexins brassilexin and sinalexin in Leptosphaeria maculans: isolation and synthesis of the elusive intermediate 3 formylindolyl-2-sulfonic acid. AB - Brassilexin and sinalexin are among the most potent antifungal phytoalexins produced by crucifer plants. Nonetheless, the crucifer pathogen Leptosphaeria maculans(Desm.) Ces. et de Not. (asexual stage Phoma lingam(Tode ex Fr.) Desm.) is able to detoxify these phytoalexins. To understand and control these reactions, the elusive brassilexin and sinalexin detoxification pathways in L. maculans were investigated and compared to those of the synthetic derivatives 8 methylbrassilexin and 8-acetylbrassilexin. The chemical structures, syntheses and antifungal activities of the metabolic intermediates, including the most polar metabolite identified thus far, 3-formylindolyl-2-sulfonic acid, were established. Our overall findings suggest that the first detoxification step of brassilexin involves its reductive bioconversion to 3-aminomethyleneindole-2 thione, followed by hydrolysis and oxidation to the water soluble metabolite 3 formylindolyl-2-sulfonic acid. PMID- 15889185 TI - Towards functionalized poly(terthiophenes): regioselective synthesis of oligoether-substituted bis(styryl)sexithiophenes. AB - A variety of new bis(oligo(oxyethylene)styryl)sexithiophenes have been prepared by chemical oxidation of ether-substituted styrylterthiophenes with FeCl3. In all cases dimers are formed in high yields, rather than the expected polymers. In addition, although three different regioisomers can potentially be formed from such an oxidation, the isolable products are shown to consist of only the head-to head regioisomer. Theoretical calculations on alkoxystyrylterthiophenes show that this can be understood in terms of an uneven electron spin density distribution at the two alpha-positions available for polymerization. Electron density calculations on the resulting head-to-head alkoxystyrylsexithiophenes show that the spin density is concentrated in the core of the molecule rather than at the alpha-positions, a result that explains the absence of significant amounts of higher oligomers. PMID- 15889186 TI - Design and synthesis of new bicyclic diketopiperazines as scaffolds for receptor probes of structurally diverse functionality. AB - Diketopiperazines (DKPs) are a common motif in various biologically active natural products, and hence they may be useful scaffolds for the rational design of receptor probes and therapeutic agents. We constructed a new bicyclic scaffold that combines a DKP bridged with a 10-membered ring. In this way we obtained a three-dimensional molecular skeleton, with several amendable sites that provide a starting point to design a new combinatorial library having diverse substituent groups. Structural variation is based upon the flexibility of alkylation of the nitrogen atoms of the DKP and on the side-chain olefin. We obtained a 10-membered secondary ring through a ring-closure metathesis reaction using the second generation Grubbs catalyst. Rings containing both O-ethers and S-ethers were compared. N-Alkyl or arylalkyl groups were introduced optionally at the two Nalpha-atoms. This is a general scheme that will allow us to test rings of varying sizes, linkages, and stereochemical parameters. The DKP derivatives were tested for activity in astrocytoma cells expressing receptors coupled to phospholipase C. Inhibitory effects were observed for signaling elicited by activation of human nucleotide P2Y receptors but not m3 muscarinic receptors. Compound 20 selectively inhibited calcium mobilization (IC50 value of 486 +/- 16 nM) and phosphoinositide turnover elicited by a selective P2Y1 receptor agonist, but this compound did not compete for binding of a radiolabeled nucleotide competitive receptor antagonist. Therefore, the new class of DKP derivatives shows utility as pharmacological tools for P2Y receptors. PMID- 15889187 TI - (S,S)-(+)-pseudoephedrine as chiral auxiliary in asymmetric acetate aldol reactions. AB - The asymmetric acetate-type aldol reaction using (S,S)-(+)-pseudoephedrine has been studied in detail. Experimental variables like the nature of the metal counterion of the enolate, the presence of additives and the structure of the aldehyde have been examined in order to reach to the highest possible yields and diastereoselectivities. PMID- 15889188 TI - Synthesis of a 4,5-epoxy-2-cyclohexen-1-one derivative via epoxide ring opening, 1,3-carbonyl transposition and epoxide ring regeneration: a synthetic study on a scyphostatin analogue. AB - A 6-alkyl-4,5-epoxy-6-hydroxy-2-cyclohexen-1-one derivative, a model compound for the hydrophilic moiety of scyphostatin, was stereoselectively synthesized from the Diels-Alder adduct. The key steps were the reductive cleavage of the 4,5 epoxide ring of the epoxidated adduct, the 1,3-carbonyl transposition of the 3 carbonyl group to the C1 position by a Wharton reaction and stereoselective bromination to provide a trans bromohydrin derivative, a precursor to the desired compound. Desilylation of the bromohydrin derivative with TBAF directly gave the target compound. PMID- 15889189 TI - Routine reoperative off-pump coronary artery bypass grafting via midline sternotomy: is it feasible? AB - Multivessel off-pump coronary artery bypass (OPCAB) has been conducted safely; however, the clinical outcomes of reoperative OPCAB have not been reported clearly. Currently OPCAB is being performed systematically in this institution including reoperative surgeries. Between September 1, 1999, and December 31, 2003, reoperative OPCAB via midline sternotomy was planned for nonemergent patients with normal left ventricular function without valvular disease. The perioperative results of patients who underwent reoperative OPCAB were analyzed. Thirty-five patients were scheduled for OPCAB. Of these, OPCAB was performed in 27 patients, giving OPCAB success rate of 77.1%. Failure of OPCAB was due to graft injury in 2, ischemic EKG change in 1, and dense adhesion and difficulty of dissection in 5 patients. The mean number of grafts with completed OPCAB was 3.0 +/- 1.5. There were no hospital deaths, strokes, or respiratory or renal failures. One patient (3.7%) developed perioperative myocardial infarction. Postoperative angiography was obtained in 12 patients (26 distal anastomoses), and all anastomoses were perfect except for 1 distal anastomosis stenosis, giving an overall stenosis-free patency rate of 95.1%. During a follow-up period (mean 2.5 +/- 0.6 years), all patients survived without cardiac event, except 1 patient who underwent catheter intervention. Cardiopulmonary bypass may be required during cardiac dissection to decompress the heart and to facilitate dissection. However, once lysis of the adhesion is completed, reoperative OPCAB can be performed without increasing surgical risks. Midterm results were satisfactory. PMID- 15889190 TI - Increased fasting plasma insulin concentrations are associated with the severity of angiographic coronary artery disease. AB - Fasting plasma insulin concentrations were obtained in 82 patients (51 men and 31 women), mean age 60 +/- 11 years, with a body mass index >25 kg/m(2) who had coronary angiography because of suspected symptomatic coronary artery disease (CAD). Obstructive CAD was diagnosed if there was >50% obstruction of > or =1 vessel. Of 82 patients, 37 (45%) had left main or 3-vessel CAD, 22 (27%) had 2 vessel CAD, 9 (11%) had 1-vessel CAD, and 14 (17%) had no obstructive CAD. Among the 4 groups, there was no significant difference in gender, age, dyslipidemia, and smoking. Hypertension (p = 0.0003), diabetes mellitus (p = 0.035), and increased fasting plasma insulin concentration (p < 0.0001) were significantly associated with the severity of CAD. Stepwise ordinal logistic regression analysis identified increased fasting plasma insulin concentrations in these obese subjects as a significant independent risk factor for the severity of angiographic CAD (p < 0.0001). PMID- 15889191 TI - Homocysteine, lipid profile, nitric oxide, vitamin B12, and folate values in patients with premature coronary artery disease and their children. AB - The plasma concentrations of homocysteine and lipoprotein A are independent risk factors for atherosclerotic vascular disease. Nitric oxide (NO) and folate values are also important in atherogenesis. The authors aimed to evaluate these parameters in patients having coronary artery bypass surgery (CABS) before 50 years of age and in their children. In 31 patients having CABS, 47 children of these patients, and 28 normal control subjects, homocysteine, NO, vitamin B12, folate, lipoprotein A, triglyceride, total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, apolipoprotein A1, and apolipoprotein B values were determined. Homocysteine values of the patients with premature coronary heart diseases and their children were significantly higher than those of controls (p < 0.031 and p < 0.006, respectively). Also, NO levels were significantly higher in both groups than in controls (p < 0.001 and p < 0.031, respectively). B12 values were significantly higher in both groups (p < 0.05 and p < 0.033, respectively). Lipoprotein A levels were higher in both groups but not significantly so. PMID- 15889192 TI - Significance of plasma nitric oxide/endothelial-1 ratio for prediction of coronary artery disease. AB - Vascular tone is regulated by vasodilators and vasoconstrictors. Endothelin-1 (ET 1) is the predominant vasoconstrictor peptide that constricts vascular smooth muscle, whereas nitric oxide (NO) is the primary vasodilator peptide that relaxes vascular smooth muscle. In this study, the authors examined whether NO/ET-1 ratio is a useful marker for detecting coronary artery disease (CAD), by comparison with evaluation based on vascular endothelial (VE) function. They measured plasma NOX and ET-1 by using ENO-200 and radioimmunoassay, in 38 subjects with normal (NL) coronary arteries (NL group; mean age, 60 +/-12 years) and 25 subjects with CAD (CAD group; mean age, 69 +/- 6 years). VE function (randomized endothelium dependent [D] and endothelium-independent [I] VE function) was assessed by measuring brachial artery (BA) diameter by using high-resolution ultrasound (7.5 MHz). Soon after these procedures, symptom-limited exercise testing was performed. There were no statistically significant differences in serum lipid concentrations or VED function between the groups. However, the CAD group had a significantly lower NO/ET-1 ratio (1.2 +/- 1.1 vs 2.7 +/- 2.2, p < 0.01) and BA diameter after sublingual nitroglycerin (VEID function: 6 +/- 7% vs 10 +/- 4%, p < 0.05). As expected, the ST segment and treadmill exercise duration were significantly lower in the CAD group. Sensitivity and specificity for detecting CAD by plasma NO/ET-1 ratio (> or =2 .0) were 90% and 85%, respectively; sensitivity and specificity for detecting CAD by ST depression (> or =1 mm) were 80% and 78%, respectively. The present results suggest that plasma NO/ET-1 ratio is a useful biological marker for predicting CAD. PMID- 15889193 TI - Exercise-induced QRS amplitude changes in patients with isolated myocardial bridging: a marker of myocardial ischemia. AB - Myocardial bridging (MB) of coronary arteries has been considered as an incidental angio-graphic finding; however, several reports suggest its association with angina pectoris, myocardial ischemia, and even infarction. In this study the authors aimed to assess exercise-induced QRS changes in patients with isolated MB and to compare those with coronary artery disease (CAD) and healthy subjects. The study population consisted of 17 patients with angiographically proven MB (group 1), 16 patients with left anterior descending (LAD) artery stenosis (group 2), and 14 healthy subjects (group 3). Each subject underwent treadmill exercise testing according to Bruce protocol. In each subject amplitude of the Q, R, and S waves in leads aVF and V(5) was measured manually before and immediately after exercise. The Athens QRS score was calculated by subtracting the Q-, R-, and S-wave differences in leads aVF and V(5). Baseline characteristics of each group were similar. There was no difference among the groups with respect to exercise testing parameters (peak heart rate, blood pressure, test duration, etc). In group 1, ST-segment depression ratio was found to be higher than that of group 3 but lower than that of group 2. In group 1, exercise QRS score was found to be lower than that of group 3 while it was higher than that of group 2 (2.9 +/- 2.3 vs 6.5 +/- 3.2 p = 0.001 and 2.9 +/- 2.3 vs 2.6 +/- 2.4 p = 0.001, respectively). In patients with MB exercise, QRS score was significantly lower than in those with normal coronary flow while it was higher in those with CAD. This may result from exercise-induced ischemia at the area perfused by the bridged artery. PMID- 15889194 TI - Increased levels of soluble adhesion molecules E-selectin and P-selectin in patients with cardiac syndrome X. AB - The role of endothelial dysfunction and platelet activation in patients with cardiac syndrome X is controversial. The aim of this study was to investigate the plasma levels of circulating E- and P-selectin molecules in patients with syndrome X. The study included 21 patients with cardiac syndrome X (11 men and 10 women, mean age = 56 +/- 5 years) and 20 patients with significant coronary artery disease who had stable angina pectoris (11 men and 9 women, mean age = 60 +/- 8 years). Twenty-two age- and sex-matched subjects (12 men and 10 women, mean age = 58 +/- 8 years) undergoing diagnosis of atypical chest pain in whom coronary arteries were found normal and exercise test had no signs of ischemia served as the control group. Syndrome X was defined as presence of typical chest pain on exertion or at rest with positive exercise test and angiographically normal epicardial coronary arteries with no evidence of coronary spasm after intracoronary infusion of ergonovine maleate. The mean plasma concentrations of P selectin were significantly elevated both in patients with coronary artery disease and syndrome X as compared with control subjects (49.15 +/-7.47 and 42.80 +/- 8.93 vs 22.63 +/-6.47 ng/mL, p < 0.001). Similarly, both patients with coronary artery disease and syndrome X had higher plasma concentrations of E selectin than the control group (78.85 +/- 16.69 and 68.38 +/- 15.30 vs 36.43 +/- 4.72 ng/mL, p < 0.001). In conclusion, patients with syndrome X had increased plasma concentrations of soluble adhesion molecules, E-selectin and P-selectin, reflecting an ongoing chronic inflammation involved with endothelial dysfunction and enhanced platelet activation/damage in this setting. PMID- 15889195 TI - Risk modification in patients with peripheral arterial disease: a retrospective survey. AB - Peripheral arterial disease (PAD) is underdiagnosed and undertreated. This is despite the high vascular morbidity and mortality rates associated with PAD. There is also evidence that quitting smoking, improving the lipid profile, lowering the blood pressure, and administering antiplatelet drugs reduce the risk of vascular events in these patients. Secondary prevention for patients with PAD is yet to meet the standard of care for those with ischemic heart disease. The authors surveyed 200 claudicants attending a vascular clinic with additional follow-up in a risk modification clinic. After a median follow-up of 28 months (range: 6-65) there was a significant (p = 0.001) improvement in walking distance; 34 patients (17%) had a vascular ischemic event. Of those, 11 patients (5.5%) had worsening intermittent claudication and 9 had a stroke/transient ischemic attack; 9 events (4.5%) were fatal. The lipid targets were met in 76% the patients. Half the smokers quit smoking and 94% of the patients were taking antiplatelet drugs or anticoagulants. Blood pressure reached the accepted target in 87% of the patients. Secondary prevention in patients with PAD may reduce the risk of vascular events. Aggressive risk modification is therefore recommended. PMID- 15889196 TI - Prevention of edema and flight microangiopathy with Venoruton (HR), (0-[beta hydroxyethyl]-rutosides) in patients with varicose veins. AB - The aim of this open study was the evaluation of the effects of HR (Venoruton) at a dose of 1 g/day on the prevention and control of flight microangiopathy and edema in subjects with varicose veins and moderate chronic venous insufficiency flying for more than 11 hours. Patients with varicose veins, edema, but without initial skin alterations or complications, were included. Measurements of skin laser Doppler (LDF) resting flux (RF) venoarteriolar response (VAR), ankle swelling (RAS), and edema were made within 12 hours before and within 3 hours after the flights. The resulting edema after the flights was evaluated with a composite edema score (analogue scale line). A group of 20 subjects was treated with HR (1 g/day, starting 2 days before the flight and 1 g for every 12 hours on day of travel). Another group of 18 subjects formed the control group. The length of the flights was between 11 and 13 hours; all seats were in coach class. Fifty patients were enrolled and 38 patients were evaluable at the end of the trial. The 2 groups (treatment and control) were comparable for age and sex distribution. The decrease in RF was significant in both groups with a higher flux at the end of the flight in the HR group (p < 0.05). The venoarteriolar response was decreased at the end of the flights; the decrease was lower in the HR group (p < 0.05). The increase in RAS and the edema score were significantly lower in the HR group. In conclusion HR is useful for reducing the level of microangiopathy and the increased capillary filtration and in controlling edema in patients with venous disease in long flights. The higher level of flux and VAR and the reduction in edema indicate a positive effect of HR on the microcirculation. This study confirms that HR prophylaxis is effective to control flight microangiopathy associated with edema. PMID- 15889197 TI - Aminophylline: exploring cardiovascular benefits versus medical malcontent. AB - Although aminophylline/theophylline has been relegated to third- and fourth-line status in the cardiopulmonary armamentarium, its use in specific pathophysiologic states, especially those of cardiac etiology, can be of significant benefit. The consulting clinician should maintain an awareness of its potential as adjunctive therapy in cases of atrioventricular block, cardiac arrest, heart failure, and bradyarrhythmias in particular. It should not yet be shelved as an archaic agent. PMID- 15889198 TI - Heparin-induced thrombocytopenia--a potential iatrogenic complication for patients with cardiac and vascular disease. AB - Physicians involved in the care of patients with cardiac and vascular problems regularly employ heparin in treatment regimens of inpatients and outpatients. Heparin-induced thrombocytopenia is an important complication that can result in significant morbidity and mortality. It is reasonable, therefore, to review the pathogenetic mechanisms involved, the identification of heparin-induced thrombocytopenia, and the management to minimize morbidity and mortality. PMID- 15889199 TI - Cardiovascular effects of type 1 diabetes mellitus in children. AB - Previous studies have shown that type 1 diabetes mellitus (DM) is associated with cardiovascular abnormalities. Early detection and treatment of these abnormalities may help to prevent the natural progression of the disease. The present study was undertaken to define early cardiovascular abnormalities in children with type 1 DM. Simultaneous evaluation of multiple cardiovascular parameters was performed in 14 children with type 1 DM and 14 age-and gender matched normal subjects. Measurements of carotid artery intima-media thickness (cIMT, echocardiography), carotid and aortic (ascending and abdominal) distensibility (echocardiography, brachial artery blood pressure), aortic pulse wave velocity (carotid to femoral artery, Doppler), and left ventricular dimensions, mass, and function (echocardiography) were performed. Diabetic children demonstrated a greater cIMT (0.36 +/- 0.04 mm vs 0.31 +/- 0.03 mm, p = 0.002) and decreased carotid artery distensibility (4.4 +/- 1.6 cm(2) . dynes(-1) . 10(-6) vs 6.0 +/- 1.9 cm(2) . dynes(-1) .10(-6), p < 0.01) compared to control. Aortic pulse wave velocity was increased in DM (6.70 +/- 0.39 vs 6.30 +/- 0.31, p = 0.02) compared to control. Left ventricular diameters, mass, and systolic and diastolic function did not differ between the 2 groups. Simultaneous assessment of multiple cardiovascular parameters in children with type 1 DM revealed impaired carotid artery structure and function, and decreased elastic properties of the aorta, before demonstrable changes in left ventricular structure and function could be detected. PMID- 15889200 TI - Left ventricular Doppler characteristics in first-degree relatives of patients with hypertrophic cardiomyopathy. AB - Left ventricular (LV) diastolic function may be affected early in patients with hypertrophic cardiomyopathy (HCM), regardless of the phenotypic expression of the disease. The aim of the present study was to detect whether LV diastolic performance, evaluated by conventional Doppler echocardiography, is impaired in first-degree relatives of patients with phenotypically expressed HCM, who had no clinical, electrocardiographic, or echocardiographic signs of the disease. Twenty two young adults having the previously described characteristics comprised the study population and 22 sex- and age-matched healthy individuals served as controls. The 2 groups were compared according to several echocardiographic parameters and the following diastolic function indices: peak velocity of E wave, representing early filling; peak velocity of A wave, representing late filling; ratio of peak early to peak late velocity (E/A); deceleration time of E wave; and LV isovolumic relaxation time. Slower deceleration time of transmitral early filling in first-degree relatives of patients with HCM (192 +/- 31 vs 149 +/- 31 msec, p < 0.001) was the only variable that significantly differentiated the 2 groups. This study shows that in healthy persons with a family history of HCM, Doppler-derived mitral filling pattern shifted toward that observed in HCM and the slower deceleration time may serve as an early sign of disease development. PMID- 15889201 TI - Feasibility of dual-chamber (DDD) pacing via a single-pass (VDD) pacing lead employing a floating atrial ring (dipole): case series, future considerations, and refinements. AB - The objective of this study was to assess the feasibility of DDD pacing from a standard single-pass VDD pacemaker system. Over the past 2 decades significant advances have been made in the development of single-pass VDD pacing systems. These have been shown in long-term prospective studies to effectively preserve atrioventricular (AV)synchrony in patients with AV block and normal sinus node function. What remains problematic is the development of a single-pass pacing system capable of DDD pacing. Such a lead configuration would be useful in those patients with peripheral venous anomalies and in younger patients with congenital anomalies, which may require lead revisions in the future. In addition, with the increased use of resynchronization (biventricular pacing) therapy, the availability of a reliable single-pass lead will minimize operative time, enhance patient safety, and minimize the amount of hardware within the heart. The feasibility of DDD pacing via a Medtronic Capsure VDD-2 (Model #5038) pacing lead was evaluated. Twenty patients who presented with AV block and normal sinus node function were recruited for this study. Atrial pacing thresholds and sensitivities were assessed intraoperatively in the supine position with various respiratory maneuvers. Five patients who agreed to participate in long-term follow-up received a dual-chamber generator and were evaluated periodically over a 12-month period. Mean atrial sensitivity was 2.35 +/- 0.83 mV at the time of implantation. Effective atrial stimulation was possible in all patients at the time of implantation (mean stimulation threshold 3.08 +/- 1.04 V at 0.5 ms [bipolar], 3.34 +/- 0.95 V at 0.5 ms [unipolar]). Five of the 20 patients received a Kappa KDR701 generator, and atrial electrical properties were followed up over a 1-year period. There was no significant change in atrial pacing threshold or incidence of phrenic nerve stimulation over the 1-year follow-up. A standard single-pass VDD pacing lead system was capable of DDD pacing intraoperatively and during long-term follow-up. Despite higher than usual thresholds via the atrial dipole, pacemaker telemetry revealed < 10% use of atrial pacing dipole over a 12-month period, which would minimally deplete the pacemaker's battery. In addition, the telemetry confirmed appropriate sensing and pacing of the atrial dipole throughout the study period. At this time such systems can serve as back-up DDD pacing systems with further refinements required to optimize atrial thresholds in all patients. PMID- 15889202 TI - Appearance of collateral circulation in the absence of total occlusive spastic coronary artery: a case report. AB - Development of coronary collateral vessels is described in a patient with coronary artery spasm associated with ST-segment depression. The collateral vessels disappeared after spasm was relieved and the ST segment was normalized, suggesting that the collateral vessels occurred during coronary artery spasm and were localized to the perfused ischemic area to prevent transmural myocardial ischemia. PMID- 15889203 TI - Unique spontaneous unhealed chronic multivessel coronary artery dissection in an elderly man: a case report and review of the literature. AB - Spontaneous coronary artery dissection (SCAD) is relatively uncommon. It often occurs in healthy, middle-aged women without overt risk factors for atherosclerosis and is associated with the peripartum period. The pattern and severity of presentation are variable. There are only 5 reports in the literature about multivessel involvement. The etiopathogenesis of SCAD is unclear. Treatment strategy is not standard but is usually tailored to clinical features. Long-term survival of patients with SCAD who have had no problem at the initial event is good. The authors report the first case of unhealed chronic multivessel SCAD in the literature. PMID- 15889204 TI - Angina-like chest pain and syncope as the clinical presentation of left ventricular endomyocardial fibrosis: a case report. AB - A 47-year-old woman complained of angina-like chest pain, near-syncope, and syncopal episodes of 17 years' duration. Physical examination was unremarkable. A 12-lead resting ECG showed symmetrically inverted T waves in the inferior and anterolateral leads. A graded treadmill exercise stress test precipitated angina like chest pain accompanied by a near-syncopal episode associated with a systemic arterial pressure of 60/40 mm Hg. Echocardiography disclosed left ventricular apical obliteration. Left ventriculogram showed a typical "ace of heart'' shadow as well as filling defects and apical obliteration. Endomyocardial biopsy of the left ventricle diagnosed left ventricular endomyocardial fibrosis. Thus, angina like chest pain and near-syncopal episodes should be added to the list of clinical manifestations of pure left ventricular endomyocardial fibrosis. PMID- 15889205 TI - Rare association of diffused coronary ectasia and anomalous origin of left circumflex coronary artery in a man with heterozygous familial hypercholesterolemia: a case report. AB - Coronary artery ectasia (CAE) is an uncommon form of coronary artery disease. It has been reported in association with a variety of pathological conditions, such as isolated congenital heart disease and Kawasaki disease. CAE is more relevant in young adults with multiple predisposing risk factors, especially familial hypercholesterolemia, and is usually considered a form of atherosclerotic coronary artery disease. A case of CAE is reported with familial hypercholesterolemia and diffuse ectasia of the coronary vessels in association with anomalous origin of the left circumflex coronary artery, which lacked ectatic segments. This combination has not been reported previously. PMID- 15889206 TI - Renal artery stenosis emerged after angiotensin-converting enzyme inhibitor treatment for myocardial infarction: a case report. AB - A 76-year-old woman with acute myocardial infarction underwent percutaneous coronary angioplasty followed by treatment with an angiotensin-converting enzyme (ACE) inhibitor, lisinopril. Her renal function deteriorated after the administration of lisinopril, so it was changed to another ACE inhibitor, temocapril. Renography suggested a complication of severe right renal artery stenosis, and renal angiography revealed bilateral renal artery stenoses. Her renal hemodynamics were assessed by (99m)Tc-Mercaptoacetyltriglycine ((99m)Tc MAG(3))-renography before and after withdrawal of temocapril. The authors concluded the patient had essential hypertension complicated by atherosclerotic renovascular disease. In the treatment of elderly patients with heart disease, hypertension, or both, with ACE inhibitor, the possibility of coexisting renal artery stenosis should be considered. Renography is recommended as a reliable tool for detecting renal artery stenosis. PMID- 15889207 TI - Profound thrombocytopenia associated with tirofiban: case report and review of literature. AB - Platelet glycoprotein (GP)IIb/IIIa inhibitors prevent fibrinogen binding and platelet aggregation. Inhibition of platelet activity at the injured coronary plaque is a target for novel therapeutic strategies. They decrease ischemic complications associated with non-ST-segment elevation acute coronary syndromes and percutaneous coronary intervention. Thrombocytopenia is a serious complication well described with the use of the prototype GP IIb/IIIa inhibitor abciximab. Its association with other agents of this class has been underemphasized. It is important to monitor platelet counts closely after initiation of GP IIb/IIIa inhibitor therapy, not only for abciximab, but also for small molecule inhibitors such as eptifibatide and tirofiban. Monitoring of platelet counts at 2 to 6 hours and 24 hours will detect most cases of acute thrombocytopenia. Adverse events may be prevented by prompt discontinuation of GP IIb/IIIa inhibitor therapy. The authors present a case of profound thrombocytopenia after the administration of tirofiban in the treatment of a patient with an acute coronary syndrome. PMID- 15889208 TI - Ascites and other extracardiac manifestations associated with right atrial myxoma: a case report. AB - Right atrial myxomas are rare intracardiac tumors that often pose difficulties in diagnosis. Right ventricular failure and ascites ensuing from tricuspid valve orifice obstruction are potentially dangerous complications. Early diagnosis of cardiac myxoma is important since surgical treatment leads to disappearance of all symptoms with a low rate of recurrence and good long-term survival. Nonspecific extracardiac symptoms, signs, complications, and laboratory findings may be the initial manifestations contributing to misdiagnosis of these rare but totally treatable atrial tumors. PMID- 15889209 TI - Transobturator SAFYRE sling is as effective as the transvaginal procedure. AB - INTRODUCTION: SAFYRE is a monofilament polypropylene mesh held between two self anchoring silicone columns that associates universal approach with readjustability. This unique feature allows for comparing two different approaches, i.e., transvaginal and transobturator, using the same sling. MATERIALS AND METHODS: A total of 226 patients with clinical and urodynamic diagnosis of SUI underwent SAFYRE sling procedure, which was performed either by transvaginal (group 1; 126 patients) or transobturator approach (group 2; 100 patients). The mean age was 63 years, in group 1 and 61 years in group 2. Physical examination, stress and pad test and urodynamic assessment were performed before the surgery. RESULTS: The average follow-up period was 18 months in group 1 and 14 months in group 2. There was no difference in cure rate in both groups. The mean operative time was longer (25 min) in group 1 than in group 2 (15 min) (P<0.05). Bladder injury was significative greater in transvaginal group [respectively, 12/126 (0%) versus 0/100 (0%)]. Postoperatively, 20.6% of the patients presented transient irritative voiding symptoms in group transvaginal group as opposed to 10% in transobturator group. DISCUSSION: SAFYRE sling performed by transobturator approach is as effective as the transvaginal procedure. Fewer complications and less operative time were additional advantages of the transobturator approach. PMID- 15889210 TI - Comparative study of surgical margins in oncoplastic surgery and quadrantectomy in breast cancer. AB - BACKGROUND: Oncoplastic surgery for breast cancer is a novel concept that combines a plastic surgical procedure with breast-conserving treatment to improve the final cosmetic results. The aim of this study was to evaluate the oncological safety of oncoplastic procedures by studying the status of the surgical margins of the excised tumor specimen in comparison with standard quadrantectomies. METHODS: Thirty consecutive breast cancer patients undergoing oncoplastic surgery (group 1) and 30 patients undergoing standard quadrantectomy (group 2) were prospectively studied with regard to the stage of breast cancer, the surgical procedures performed, the volume of breast tissue excised, and the histopathology of the tumor specimen, with specific details on surgical margins. RESULTS: Patients who underwent oncoplastic surgery (group 1) were younger (mean age, 48.73 years) than patients who had a classic quadrantectomy (group 2; mean age, 55.76 years; P = .022). The mean volume of the excised specimen in group 1 was 200.18 cm(3), compared with 117.55 cm(3) in group 2 (P = .016). Surgical margins were negative in 25 cases out of 30 in group 1 and 17 out of 30 in group 2 (P = .05). The average length of the surgical margin was 8.5 mm in group 1 and 6.5 mm in group 2, but the difference was not statistically significant (P = .074). CONCLUSIONS: Oncoplastic surgery adds to the oncological safety of breast conserving treatment because a larger volume of breast tissue can be excised and a wider negative margin can be obtained. It is especially indicated for large tumors, for which standard breast-conserving treatment has a high probability of leaving positive margins. PMID- 15889211 TI - Fusion image of positron emission tomography and computed tomography for the diagnosis of local recurrence of rectal cancer. AB - BACKGROUND: The aim of this study was to evaluate the clinical and therapeutic value of digital fusion image (FI) of positron emission tomography (PET) using (18)F-fluorodeoxy glucose and computed tomography (CT) in patients who were suspected of having a local recurrence of rectal cancer. METHODS: Forty-two patients (32 men and 10 women; mean age, 61.4 years, range, 40-79 years) with a suspicion of local recurrence after curative resection of rectal cancer were prospectively recruited and underwent (18)F-fluorodeoxy glucose-PET and CT. The FI was reconstructed with a commercially available digital software program, T-B Fusion. Wilcoxon signed rank test was used to compare FI with CT alone or PET alone. RESULTS: FI yielded a correct diagnosis in 39 (93%) of 42 patients, whereas CT alone and PET alone did so in 33 (79%) and 37 (88%) patients, respectively. FI had better diagnostic accuracy than CT alone (P = .0138) and PET alone (P = .0156). Overall, FI altered patient management in 11 (26.2%) patients on the basis of additional information, including differentiation of the tumor from the postoperative scar in 6 patients, exact anatomical location in 3 patients, and both in 2 patients. CONCLUSIONS: FI has a potential clinical value in the treatment of suspected local recurrence of rectal cancer. PMID- 15889212 TI - The impact of nonvisualization of sentinel nodes on lymphoscintigraphy in breast cancer. AB - BACKGROUND: This study aimed at evaluating the relationship between the nonvisualization of sentinel nodes (SNs) at lymphoscintigraphy and the intraoperative detection rate, radioactive counts in vivo, and histological status of SNs. METHODS: Two hundred eighty patients with infiltrating breast carcinoma (T0, T(1)/T(2)) underwent preoperative lymphoscintigraphy before gamma probe-guided SN biopsy. RESULTS: The surgical identification rate with a gamma probe was 84.6% (56 of 280) in lymphoscintigraphy-negative patients and 93.2% (224 of 280) in lymphoscintigraphy-positive patients (P < .05) after two subdermal periareolar injections. The average number of SNs per patient was 1.7 in lymphoscintigraphy-negative patients and 2.2 in lymphoscintigraphy-positive patients (P < .01), as assessed by gamma detection. The mean age of lymphoscintigraphy-negative patients was 62 +/- 10 years, versus 55 +/- 13 years for lymphoscintigraphy-positive patients (P < .001). The median radioactive count in dissected SNs identified by gamma detection was 204 cps (range, 4-618 cps) in lymphoscintigraphy-negative patients, versus 606 cps (range, 43-16,928 cps) in lymphoscintigraphy-positive patients (P < .001). The rate of macrometastatic SNs was 40% in lymphoscintigraphy-negative patients, versus 30% in lymphoscintigraphy positive patients (not significant), whereas the size of involved SNs was 16.6 mm in lymphoscintigraphy-negative patients, versus 13.1 in lymphoscintigraphy positive patients (P < .05). The micrometastasis detection rate in SNs from lymphoscintigraphy-negative patients was 6.25%, versus 23.3% in lymphoscintigraphy-positive patients (P < .01). CONCLUSIONS: Negative lymphoscintigraphy was observed in 20% of patients and was more frequent in elderly patients. Negative lymphoscintigraphy was predictive of a lower surgical identification rate and fewer detected SNs. These SNs had fewer micrometastases, were fairly large, and tended to harbor metastases. PMID- 15889213 TI - A tat fusion protein-based tumor vaccine for breast cancer. AB - BACKGROUND: We recently reported that dendritic cells (DCs) transduced with a fusion protein between Her2/neu and the protein transduction domain Tat (DC-Tat extracellular domain [ECD]) induced Her2/neu-specific CD8(+) T cells in vitro. This study tested the in vivo efficacy of DC-Tat-ECD in a murine breast cancer model. METHODS: FVB/N mice received one or two weekly intraperitoneal immunizations with syngeneic DC-Tat-ECD followed by a tumor challenge with syngeneic neu(+) breast cancer cells, and tumor development was monitored. To test for Her2/neu specificity, CD4(+) and CD8(+) cells were isolated through magnetic bead separation and analyzed for specific interferon gamma release. RESULTS: Intraperitoneally injected DCs migrated to secondary lymphoid organs, as evidenced by small-animal positron emission tomography studies. Immunized mice developed palpable tumors significantly later than control mice injected with DC Tat-empty (P = .001 and P < .05 for two immunizations and for one immunization, respectively) or mice that received no DCs (P = .001 and P < .05). Similarly, immunized mice had smaller resulting tumors than mice injected with DC-Tat-empty (P < .05 and P < .01) or untreated mice (P < .001 and P < .001). Significantly more tumor-specific CD8(+) splenocytes were found in twice-immunized mice than in untreated animals (P < .001). Similarly, a T-helper type 1 CD4(+) T-cell response was observed. CONCLUSIONS: Protein-transduced DCs may be effective vaccines for the treatment of cancer. PMID- 15889214 TI - Primary hyperparathyroidism: can parathyroid carcinoma be anticipated on clinical and biochemical grounds? Report of nine cases and review of the literature. AB - BACKGROUND: Parathyroid carcinoma (PC) mimics benign primary hyperparathyroidism (PHP), but the diagnosis of PC is seldom available at the time of the first operation. Because PC is plagued by recurrences usually beyond cure, one may wonder whether some of these could be prevented by more extensive resections initially, i.e., if the diagnosis of PC were available at that time. METHODS: Over a 25-year period, 311 consecutive patients with PHP underwent operation in our department: 302 had benign disease (adenomas or hyperplasias), and 9 had PC. Several clinical parameters, serum calcium and parathyroid hormone (PTH) levels, and the weight of the parathyroid tumor removed were compared in both groups. Receiver operating characteristic curves and logistical regression analyses were used to distinguish PC from benign PHP. RESULTS: Eight of 9 patients with PC had symptoms, versus 238 (79%) of 302 with benign PHP (not significant). In the PC subgroup, serum calcium and PTH levels and the tumor weights of the parathyroid glands removed were significantly higher than in the benign PHP cohort, even if these three parameters were regularly flawed by low positive predictive values (14%, 20%, and 15%, respectively). CONCLUSIONS: Serum calcium, PTH levels, and tumor weights were significantly greater in the PC subgroup, even if not invariably in a discriminatory way. However, when PTH is <4 times the upper limit of normal and tumor weight is <1.9 g, the probability of PC is nil. PMID- 15889215 TI - External jugular vein cutdown approach, as a useful alternative, supports the choice of the cephalic vein for totally implantable access device placement. AB - BACKGROUND: Cephalic vein (CV) cut down for totally implantable venous access device (TIVAD) placement has been accepted as an alternative to the percutaneous subclavian vein approach. The aim of this retrospective study was to validate the external jugular vein (EJV) cut-down approach when the CV is not feasible. METHODS: Patients receiving a TIVAD from January 1995 to December 2003 were included in this study. Age, sex, surgical technique, disease, device used, length of the procedure, and morbidity were considered. RESULTS: A total of 427 TIVADs were placed in 425 patients: 253 men (59.5%) and 172 women (40.5%) aged 31 to 79 years. Of 425 patients, 5 were excluded; 420 underwent a CV cut down on the first attempt, and 391 (93.1%) procedures were successful. Among the final 29 patients, 20 (68.96%) underwent a TIVAD placement through the ipsilateral EJV cut down approach. In the remaining nine patients (31.04%), TIVAD placement was performed through the ipsilateral internal jugular vein in four cases, via the ipsilateral axillary vein in three cases, and through the ipsilateral coracobrachial vein in the other cases. No immediate postoperative complications were detected in any of the patients. CONCLUSIONS: TIVAD placement by the CV cut down approach is safe and fast, and its success rate is very high. By avoiding the immediate complications associated with the percutaneous approach, the EJV cut down has to be considered a valid, safe, and suitable alternative when the CV is not feasible. PMID- 15889216 TI - Pathologic nipple discharge: surgery is imperative in postmenopausal women. AB - BACKGROUND: A total of 10% to 15% of pathologic nipple discharge in women is due to malignant lesions of the breast. The purpose of this study was to discover the rate of breast cancer in women who present with this symptom and undergo ductal excision, to evaluate the different diagnostic methods used before surgery, and to discover whether there are specific factors with regard to dignity. METHODS: We analyzed 118 ductal excisions in 116 patients performed at the women's hospital of the University of Rostock, Germany, between 1995 and 2002. The discharging duct was identified by preoperative galactography. RESULTS: The rate of cancer in these patients was 9.3% (n = 11). The most frequent benign lesion was intraductal papillomatous proliferation (36.4%; n = 43). Solitary papillomas were shown in 21.2% (n = 25), and other specific benign histologic findings were shown in 27.1% (n = 32). Women with malignancies were significantly older (P = .009) and were more often postmenopausal (P = .095) compared with patients with benign histology. Galactography was the method that reached the highest sensitivity (73%), and clinical examination showed the highest specificity (85%) in distinguishing between benign and malignant lesions. CONCLUSIONS: Because 94.1% of all cases presented with specific histological findings causing pathologic nipple discharge, ductal excision combined with preoperative galactography was proven to be a sufficient method for diagnosis and therapy. This procedure should be performed in all postmenopausal women with this symptom because of a cancer rate of 12.7% among this age group and the unsatisfactory quality of other diagnostic methods. PMID- 15889217 TI - The outcome of surgical resection versus assignment to the liver transplant waiting list for hepatocellular carcinoma. AB - BACKGROUND: Optimal management of patients with hepatocellular carcinoma (HCC) is controversial. This study was conducted to evaluate the outcome of tumor resection versus assignment to a liver transplant waiting list (WL) in patients with HCC. METHODS: Prospectively collected patient data from 1970 to 1997 on 313 patients with HCC were retrospectively analyzed by multivariate analysis to determine the effect of liver disease, method of treatment, and tumor-related factors on survival. RESULTS: A total of 199 patients underwent nonsurgical palliative care (PC), 81 underwent partial liver resection (LR), and 33 were assigned to a liver transplant WL, of which 22 received a donor liver. A total of 91%, 53%, and 91% of the patients had cirrhotic livers in the PC, LR, and WL groups, respectively (P < .001). In the LR group, the absence of a tumor capsule (P < .0001) and a poorly differentiated tumor (P = .027) were both adverse prognostic factors. In the WL group, hepatitis B (P = .02) and American Joint Committee on Cancer tumor stage III (P = .019) were adverse prognostic factors. The 3-year survival rates were 4%, 33%, and 38% for the PC, LR, and WL patients, respectively (P < .0001). The 3-year survival rate in the LR patients was 51% in patients without cirrhosis and 15% in patients with cirrhosis (P < .0001). CONCLUSIONS: Patients with locally unresectable tumors, distant disease, or both will continue to receive PC. Patients assigned to liver transplant WLs run the risk of not receiving a donor liver, in which case their survival is predicted to be poor. Survival after resection in a group of patients with advanced tumors is worse than that after transplantation; however, shortages of donor livers presently preclude transplantation in this population of patients. PMID- 15889218 TI - Prevention of surgical site infection after open prosthetic inguinal hernia repair: efficacy of parenteral versus oral prophylaxis with amoxicillin clavulanic acid in a randomized clinical trial. AB - The aim of this prospective study was to compare the efficacy of oral versus parenteral prophylactic amoxicillin-clavulanic acid for preventing surgical site infection after open prosthetic mesh repair of inguinal hernia. A total of 480 inguinal-hernia patients were randomly assigned to two groups. Group I (n = 240) received 1.313 g oral amoxicillin-clavulanic acid 2 hours before operation, and group II (n = 240) received 1.2 g of the same drug combination intravenously approximately 30 minutes before surgery. Patients were examined four times during 1 year of follow-up (at 7-10 days, 4-6 weeks, 6 months, and 12 months postoperation), and data related to surgical site infections were collected. Seventy-two patients were excluded due to confounding factors during and after the operation. There were no statistically significant differences between group I (final n = 208) and group II (final n = 200) with respect to age, sex distribution, body mass index, American Anesthesiology Association grade, frequencies of different hernia types, duration of surgery, and the experience levels of the principal surgeon in the operations. One of the 208 (0.5%) patients in group I and 3 of the 200 (1.5%) patients in group II developed superficial surgical site infections (p > 0.05). None of the infections required mesh removal. There were no deep surgical site infections in either group, and there was one case of hernia recurrence in each group. For patients undergoing open prosthetic repair of inguinal hernia, oral amoxicillin-clavulanic acid is safe, significantly less costly, and equally effective in preventing surgical site infection as the same dose given parenterally. PMID- 15889219 TI - Regional distribution of biogenic amines, amino acids and cholinergic markers in the CNS of the C57BL/6 strain. AB - A reliable extrapolation of neurochemical alterations from a mouse model to human metabolic brain disease requires knowledge of neurotransmitter levels and related compounds in control mouse brain. C57BL/6 is a widely used background strain for knockout and transgenic mouse models. A prerequisite for reliable extrapolation from mouse brain to the human condition is the existence of analogous distribution patterns of neurotransmitters and related compounds in control mouse and human brain. We analysed regional distribution patterns of biogenic amines, neurotransmitter and non-neurotransmitter amino acids, and cholinergic markers. Distribution patterns were compared with known neurotransmitter pathways in human brain. The present study provides a reference work for future analyses of neurotransmitters and related compounds in mouse models bred in a C57BL/6 background strain. PMID- 15889220 TI - Synthesis, characteristics and biological activity of pentacoordinated spirophosphoranes derived from amino acids. AB - The reactions of phosphorus trichloride with various amino acids afford the pentacoordinated spirophosphoranes. The reaction procedures were traced by (31)P NMR spectra techniques. A new crystal structure of alanine derivative was characterized, which is a slightly distorted TBP structure. Besides, this kind of spirophosphoranes are potent inhibitors to tyrosinase. PMID- 15889221 TI - Using pseudo amino acid composition to predict protein subcellular location: approached with Lyapunov index, Bessel function, and Chebyshev filter. AB - With the avalanche of new protein sequences we are facing in the post-genomic era, it is vitally important to develop an automated method for fast and accurately determining the subcellular location of uncharacterized proteins. In this article, based on the concept of pseudo amino acid composition (Chou, K.C. Proteins: Structure, Function, and Genetics, 2001, 43: 246-255), three pseudo amino acid components are introduced via Lyapunov index, Bessel function, Chebyshev filter that can be more efficiently used to deal with the chaos and complexity in protein sequences, leading to a higher success rate in predicting protein subcellular location. PMID- 15889224 TI - Abstracts of the 1st Conference of InSiGHT, the International Society for Gastrointestinal Hereditary Tumors, Newcastle upon Tyne, United Kingdom, 14-17 June 2005. PMID- 15889226 TI - 5th European Cytogenetics Conference, June 4-7, 2005, Madrid, Spain. Abstracts. PMID- 15889229 TI - [Dermato-oncological rehabilitation]. AB - National insurance companies in Germany support health cures for patients with malignant tumors (malignant melanoma, squamous cell carcinoma, Merkel cell tumor, malignant cutaneous lymphoma). The clinical requirements are an invasively growing tumor, problems of self-assurance, and dis-integration of the patient regarding his social and/or professional environment. The decision for a health cure is made by the treating dermatologist in the hospital. In this context, the following sociomedical criteria should be applied: impairment, disability, and handicap. Usually, rehabilitation starts after the patient is discharged from the hospital. The inpatient rehabilitation program should be performed at an institution capable of providing dermatological and psychological treatment. The dermatologist acts as a manager for the members of the rehabilitation team (psychologists, physiotherapists, social workers, and ergo-therapists). In conclusion, dermato-oncologic rehabilitation plays an important role in re integrating the patient into his professional life to avoid retirement. PMID- 15889230 TI - [Lichen sclerosus. New aspects of pathogenesis and treatment]. AB - Lichen sclerosus et atrophicus is a chronic inflammatory disease that mainly affects women in the 5th decade. Although lichen sclerosus most often affects the anogenital region, it may occur in other cutaneous or mucosal sites. Increased fibroblast activity causes cutaneous sclerosis. Recent studies have identified lichen sclerosus as an autoantibody-mediated chronic inflammatory dermatosis. Autoantibodies against the extracellular matrix protein-1 are present in up to 80% of affected patients. In addition to the well-accepted therapy with potent corticosteroids, promising results have been obtained using calcineurin antagonists in the treatment of lichen sclerosus. Interdisciplinary management with regular monitoring can improve the clinical manifestations and quality of life. PMID- 15889231 TI - Regulating adiponectin: of flax and flux. PMID- 15889232 TI - Dysregulation of energy homeostasis in mice overexpressing insulin-like growth factor-binding protein 6 in the brain. AB - AIMS/HYPOTHESIS: IGFs, IGF receptors and IGF binding proteins (IGFBPs) are widely expressed in the central nervous system. To investigate the physiological significance of IGFBP-6 in the brain we established two transgenic mouse lines overexpressing human (h)-IGFBP-6 under the control of glial fibrillary acidic protein promoter. Increasing evidence suggests that insulin/IGF signalling pathways could be implicated in the neuroendocrine regulation of energy homeostasis. We explored the impact of brain IGFBP-6 overexpression on the regulation of food intake and energy balance. METHODS: Transgenic mice were fed either a control diet or a high-fat diet for up to 3 months. Glucose and insulin tolerance tests were carried out before and after the diet period. Plasma parameters (insulin, leptin, glucose, NEFAs and triglycerides) were measured, and uncoupling protein 1 (UCP-1) expression was quantified in brown adipose tissue. Oxygen consumption was also measured in both groups. RESULTS: The transgenic mice fed a high-fat diet for 3 months developed obesity, showing increases in plasma leptin, glucose and insulin levels and mild insulin resistance. As compared with wild-type mice, no significant differences were found in the quantity of food intake. However, UCP-1 expression was down-regulated in the brown adipose tissue of the transgenic mice. CONCLUSIONS/INTERPRETATION: Our results show that brain IGFBP-6 has an impact on the regulation of energy homeostasis. These transgenic h IGFBP-6 mice may be considered a new tool for studies of the involvement of the brain IGF system in metabolism control and obesity. PMID- 15889233 TI - Potential impact of American Diabetes Association (2000) criteria for diagnosis of gestational diabetes mellitus in Spain. AB - AIMS/HYPOTHESIS: This study was carried out to determine the impact of American Diabetes Association (ADA) 2000 criteria for the diagnosis of gestational diabetes mellitus (GDM) in the Spanish population. METHODS: Pregnant women were assigned to one of four categories: negative screenees, false-positive screenees, ADA-only-GDM (untreated) and GDM according to National Diabetes Data Group (NDDG) criteria (treated). Fetal macrosomia and Caesarean section were defined as primary outcomes, with seven additional secondary outcomes. RESULTS: Of 9,270 pregnant women screened for GDM, 819 (8.8%) met NDDG criteria. If the threshold for defining GDM had been lowered to ADA criteria, an additional 2.8% of women would have been defined as having the condition (relative increase of 31.8%). Maternal characteristics of women with ADA-only-GDM were between those of false positive screenees and women with NDDG-GDM. The risk of diabetes-associated complications was slightly elevated in the individuals who would have been classified as abnormal only after the adoption of ADA criteria. In addition, the ADA-only-GDM contribution to morbidity was lower than that of other variables, especially BMI. CONCLUSIONS/INTERPRETATION: Use of the ADA criteria to identify GDM would result in a 31.8% increase in prevalence compared with NDDG criteria. However, as the contribution of these additionally diagnosed cases to adverse GDM outcomes is not substantial, a change in diagnostic criteria is not warranted in our setting. PMID- 15889234 TI - Long-term efficacy and tolerability of add-on pioglitazone therapy to failing monotherapy compared with addition of gliclazide or metformin in patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: The aim of this analysis was to examine the long-term effects of pioglitazone or gliclazide addition to failing metformin monotherapy and pioglitazone or metformin addition to failing sulphonylurea monotherapy in patients with type 2 diabetes. METHODS: Two 2-year, randomised, multicentre trials were performed in patients with inadequately controlled type 2 diabetes (HbA1c 7.5-11% inclusive), who were receiving either metformin or a sulphonylurea at > or = 50% of the maximum recommended dose or at the maximum tolerated dose. In the first study, patients on metformin received add-on therapy with pioglitazone (15-45 mg/day, n = 317) or gliclazide (80-320 mg/day, n = 313). In the second study, patients on sulphonylurea therapy were randomised to receive add-on therapy with either pioglitazone (15-45 mg/day, n = 319) or metformin (850 2,550 mg/day, n = 320). HbA(1)c, fasting plasma glucose, insulin and lipids were investigated. RESULTS: At week 104, the mean reduction from baseline in HbA(1)c was 0.89% for pioglitazone and 0.77% for gliclazide addition to metformin (p = 0.200). There was a statistically significant between-group difference for the change in mean fasting plasma glucose at week 104 (-1.8 mmol/l for pioglitazone vs -1.1 mmol/l for gliclazide, p < 0.001). There were no significant differences in changes from baseline in glycaemic parameters for pioglitazone compared with metformin addition to sulphonylurea therapy. Whether added to metformin or sulphonylurea, pioglitazone caused significantly greater decreases in triglycerides and significantly greater increases in HDL cholesterol than the comparator regimens (p < or = 0.001). There were decreases in LDL cholesterol in the comparator groups and these were significantly different from the small changes observed with pioglitazone (p < 0.001). All treatment regimens were well tolerated. There were weight increases of 2.5 kg and 3.7 kg in the pioglitazone and 1.2 kg in the gliclazide add-on groups, and there was a mean decrease of 1.7 kg in the metformin add-on group. CONCLUSIONS/INTERPRETATION: As add-on therapy to existing sulphonylurea or metformin therapy, pioglitazone improved glycaemic control and this improvement was sustained over 2 years. Furthermore, there were potential benefits in terms of improvements in specific lipid abnormalities. This could offer an advantage over the addition of other oral agents in the long-term treatment of diabetes. PMID- 15889235 TI - A dietary pattern protective against type 2 diabetes in the European Prospective Investigation into Cancer and Nutrition (EPIC)--Potsdam Study cohort. AB - AIMS/HYPOTHESIS: The aim of this study was to identify a dietary pattern associated with diabetes-related biomarkers and to investigate whether this pattern is associated with the incidence of type 2 diabetes. METHODS: A nested case-control study of 192 cases of incident type 2 diabetes and 382 control subjects matched for sex and age was conducted. All subjects were participants in the population-based European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam Study. Dietary pattern score was derived using intake data on 48 food groups as exposure variables and the biomarkers HbA1c, HDL cholesterol, C reactive protein and adiponectin as response variables in reduced rank regression. The association of the score with diabetes risk was estimated by conditional logistic regression analysis. RESULTS: A high score for the identified dietary pattern was characterised by a high intake of fresh fruit and a low intake of high-caloric soft drinks, beer, red meat, poultry, processed meat, legumes and bread (excluding wholegrain bread). Subjects with high scores had high plasma concentrations of HDL cholesterol and adiponectin and low plasma concentrations of HbA1c and C-reactive protein. After multivariate adjustment, the odds ratios for type 2 diabetes across increasing quintiles of the dietary pattern score were 1.0, 0.59, 0.51, 0.26 and 0.27, respectively (p = 0.0006 for trend). CONCLUSIONS/INTERPRETATION: A high score for the identified dietary pattern is associated with a more favourable biomarker profile and a substantially reduced incidence of type 2 diabetes. PMID- 15889236 TI - The uridine diphosphate glucuronosyltransferases: quantitative structure-activity relationships for hydroxyl polychlorinated biphenyl substrates. AB - Quantitative structure-activity relationships (QSARs), which relate the glucuronidation of hydroxyl polychlorinated biphenyls (OH-PCBs)-catalyzed by the uridine diphosphate glucuronosyltransferases (UGTs)-to their physicochemical properties and molecular structural parameters, can be used to predict the rate constants and interpret the mechanism of glucuronidation. In this study, QSARs have been developed that use 23 semi-empirical calculated quantum chemical descriptors to predict the logarithms of the constants 1/K(m) and V(max), related to enzyme kinetics. A partial least squares regression method was used to select the optimal set of descriptors to minimize the multicollinearity between the descriptors, as well as to maximize the cross-validated coefficient (Q(2) (cum)) values. The key descriptors affecting log(1/K(m)) were E(lumo)- E(homo) (the energy gap between the lowest unoccupied molecular orbital and the highest occupied molecular orbital) and q(C) (-) (the largest negative net atomic charge on a carbon atom), while the key descriptors affecting logV(max) were the polarizability alpha, the Connolly solvent-excluded volume (CSEV), and logP (the logarithm of the partition coefficient for octanol/water). From the results obtained it can be concluded that hydrophobic and electronic aspects of OH-PCBs are important in the glucuronidation of OH-PCBs. PMID- 15889237 TI - Indomethacin induces free radical-mediated changes in renal brush border membranes. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are used extensively in clinical medicine. One disadvantage of their use, however, is the occurrence of adverse effects in the kidneys. The side effects produced in this organ have been classically attributed to the inhibitory effect of these drugs on the activity of cyclooxygenase, a key enzyme in prostaglandin synthesis. Our earlier work with indomethacin, a commonly used NSAID, has shown that oxidative stress and mitochondrial dysfunction occur in the kidney in response to the drug. In view of this, this study looked into the effect of indomethacin on brush border membranes (BBM) from the kidney, as these biomembranes are prime targets of oxygen free radicals. Rats, fasted overnight, were dosed with indomethacin (20 mg/kg) by gavage and sacrificed 24 h later. BBM were isolated from the kidneys by polyethylene glycol precipitation. It was found that there was an increase in levels of products of peroxidation and a fall in the level of alpha-tocopherol in the BBM from indomethacin-dosed rats. These BBM also exhibited impaired glucose transport. The lipid composition of the membranes was also found to be altered. Alterations in lipids were associated with up-regulation of phospholipase A2. Pretreatment with L-arginine, a nitric oxide donor, protected against these effects of indomethacin. Thus, this study suggests that indomethacin induces impairment in structure and function of BBM in the kidney, with these effects possibly mediated by free radicals and activation of phospholipases. We postulate that such alterations may be important in the pathogenesis of NSAID-induced nephropathy. PMID- 15889238 TI - Effect of methanol on endogenous and exogenous carnitine levels in rat plasma. AB - The effect of methanol on the levels of endogenous carnitine and its derivatives was studied in male Sprague-Dawley rats aged three months. In addition, the effect of L-carnitine supplementation on metabolic disturbances caused by methanol intoxication was studied. The rats were randomized into six groups, including two control groups. Methanol was given at 1/4 LD(50) and 1/2 LD(50)/kg b.w. (or water in control) through an intragastric tube, and L-carnitine (or 0.9% NaCl in the control) was injected intraperitoneally. The levels of plasma L carnitine and its derivatives were measured at selected time points for four days. Following methanol administration, the rats exhibited dose-dependent increases in L-carnitine levels and altered ratios of L-carnitine and its derivatives. L-carnitine supplementation accelerated the normalization of metabolic disturbances, as indicated by the acylcarnitine to free carnitine ratio (AC/FC). The protective effect of L-carnitine is supported by the fact that 100% of the methanol-treated rats supplemented with carnitine survived, while 8/60 rats and 27/101 rats died at methanol doses of 1/4 LD(50) and 1/2 LD(50), respectively, in groups without L-carnitine supplementation. PMID- 15889239 TI - TRPC3: a versatile transducer molecule that serves integration and diversification of cellular signals. AB - Transient receptor potential (TRP) proteins have been recognized as sensors for a wide variety of external and internal signals involved in maintenance of cellular homeostasis and control of physiological functions. Evidence of a striking versatility in terms of signal integration and transduction has been reported for members of the canonical (or classical) TRP subfamily (TRPCs). TRPC species are cation channel subunits and emerge as multifunctional signal transduction molecules that are able to function as components of divergent signalplexes. Results obtained in heterologous expression systems suggest TRPC3 as a paradigm of multifunctional signal transduction by a cation channel protein. TRPC3 serves cellular Ca(2+) signaling by multiple mechanisms and may control a variety of distinct physiological functions. In this review, we summarize current knowledge on the properties and possible signaling partners of TRPC3, and discuss the role of TRPC3 channel proteins in cellular signaling networks. PMID- 15889240 TI - Structure-function analysis of TRPV channels. AB - In recent years many new members of the family of TRP ion channels have been identified. These channels are classified into several subgroups and participate in many sensory and physiological functions. TRPV channels are important for the perception of pain, temperature sensing, osmotic regulation, and maintenance of calcium homeostasis, and much recent research concerns the identification of protein domains involved in mediating specific channel functions. Recent literature on TRPV channel subunit composition, protein domains required for subunit assembly, trafficking, and regulation will be reviewed and discussed. PMID- 15889241 TI - Neuronal effects of acute citalopram detected by pharmacoMRI. AB - RATIONALE: Serotonin (5-hydroxytryptamine, 5-HT) is implicated in the aetiology and treatment of a variety of psychiatric disorders. A limitation of research has been the necessity to use indirect measures of 5-HT function. METHOD: We describe a method of analysing pharmacoMRI data using SPM and apply it to the direct i.v. infusion of selective 5-HT reuptake inhibitor, citalopram, in 12 healthy volunteers. Scanning took place on a 1.5-T Philips MRI scanner. RESULTS: Areas implicated in depression and its treatment were observed to have increasing signal with respect to time. These areas included the caudate, the amygdala, the hippocampus, the striatum and the thalamus. CONCLUSION: Direct pMRI using i.v. citalopram opens new ways of investigating 5-HT mechanism in depression and its treatment. PMID- 15889242 TI - Enhancing the ratio of molecular ions to non-covalent compounds in the electrospray interface of LC-MS in quantitative analysis. AB - A common problem encountered during the development of MS methods for the quantitation of small organic molecules by LC-MS is the formation of non covalently bound species or adducts in the electrospray interface. Often the population of the molecular ion is insignificant compared to those of all other forms of the analyte produced in the electrospray, making it difficult to obtain the sensitivity required for accurate quantitation. We have investigated the effects of the following variables: orifice potential, nebulizer gas flow, temperature, solvent composition and the sample pH on the relative distributions of ions of the types MH+, MNa+, MNH4+, and 2MNa+, where M represents a small organic molecule: BAY 11-7082 ((E)-3-[4-methylphenylsulfonyl]-2-propenenitrile). Orifice potential, solvent composition and the sample pH had the greatest influence on the relative distributions of these ions, making these parameters the most useful for optimizing methods for the quantitation of small molecules. PMID- 15889243 TI - Combined solid-phase extraction and 2D LC-MS for characterization of the neuropeptides in rat-brain tissue. AB - A comprehensive two-dimensional capillary liquid chromatographic (2D LC) method has been established for determination of neuropeptides in rat brain tissue. Rats were exposed to different levels of stress before sacrificing and the aim of this study was to design a powerful separation and detection technique capable of characterizing differences between cerebral neuropeptide expression as a function of stress level. Rat brain samples were homogenized and subjected to clean-up by solid-phase extraction (SPE) on both a reversed-phase (C(18)) and a weak cation exchange (CBA) cartridge. The samples were divided in two fractions (A and B) depending on retention on the CBA column. Subsequently, 50 microL of the sample were injected on to a strong cation exchanger (SCX) at a mobile phase pH of 3, which enabled preconcentration of positively charged compounds. The trapped compounds were eluted using step gradients of ammonium formate in water-ACN (90:10, v/v). Before enrichment in the second dimension, the eluate from the first dimension was diluted with water containing 0.1% TFA. The compounds eluting from the first dimension were trapped in the second dimension using a dual precolumn system consisting of two short capillary columns packed with Kromasil C(18), 10 microm particles. Subsequently, the trapped compounds were backflushed on to a 10 cm long, 320 microm I.D. analytical column packed with Kromasil C(18) 3.5 microm particles, on which they were efficiently separated. Detection was performed using an ion-trap mass spectrometer (ITMS) in both the MS and the MS-MS mode. Comparison of base-peak chromatograms (BPC) from MS analysis of stressed and non-stressed rats clearly revealed several differences in neuropeptide expression. The MS-MS data obtained combined with Mascot software were employed for peptide identification. PMID- 15889244 TI - Vestibular and non-vestibular contributions to eye movements that compensate for head rotations during viewing of near targets. AB - Geometry dictates that when subjects view a near target during head rotation the eyes must rotate more than the head. The relative contribution to this compensatory response by adjustment of the vestibulo-ocular reflex gain (Gvor), visual tracking mechanisms including prediction, and convergence is debated. We studied horizontal eye movements induced by sinusoidal 0.2-2.8 Hz, en-bloc yaw rotation as ten normal humans viewed a near target that was either earth-fixed (EFT) or head-fixed (HFT). For EFT, group median gain was 1.49 at 0.2 Hz declining to 1.08 at 2.8 Hz. For HFT, group median gain was 0.03 at 0.2 Hz increasing to 0.71 at 2.8 Hz. By applying transient head perturbations (peak acceleration >1,000 degrees s(-2)) during sinusoidal rotation, we determined that Gvor was similar during either EFT or HFT conditions, and contributed only approximately 75% to the compensatory response. We confirmed that retinal image slip contributed to the compensatory response by demonstrating reduced gain during EFT viewing under strobe illumination. Gain also declined during sum-of sines head rotations, confirming the contribution of predictive mechanisms. The gain of compensatory eye movements was similar during monocular or binocular viewing, although vergence angle was greater during binocular viewing. Comparison with previous studies indicates that mechanisms for generation of eye rotations during near viewing depend on head stimulus type (rotation or translation), waveform (transient or sinusoidal), and the species being tested. PMID- 15889245 TI - Magnetic resonance imaging findings of proliferating trichilemmal tumor. AB - We report the magnetic resonance (MR) imaging findings of proliferative trichilemmal tumor (PTT), a rare benign tumor of the scalp. MR imaging showed a well-margined mass with isointense signal on T(1)-weighted images and heterogeneous mixed intensity signal on T(2)-weighted images After i.v. administration of contrast material, the mass showed significant enhancement, with small portions remaining unenhanced. It was considered that these findings reflected the pathological characteristics of PTT, which consists of solid lobules and cystic cavities. PMID- 15889246 TI - Bone measurements of infants in the first 3 months of life by quantitative ultrasound: the influence of gestational age, season, and postnatal age. AB - BACKGROUND: There are a few quantitative ultrasound (QUS) studies of bone status for Chinese children. OBJECTIVE: To evaluate the clinical application and to investigate the bone status of neonates and young infants with QUS. MATERIALS AND METHODS: An ultrasound bone sonometer was used to measure the bone speed of sound (SOS) of the tibia in 542 neonates within 3 months of birth. RESULTS: At birth, no significant difference of SOS was found between boys and girls, but there was a significant difference of SOS between premature infants and full-term infants. The SOS in neonates born during spring and summer was significantly lower than those born during autumn and winter. There were significant correlations between SOS and gestational age, and between bone SOS and birth weight in appropriate for gestational age (AGA) infants. Multiple regression analysis found that gestational age and infant birth season were two important factors influencing SOS. During the first 3 months, there was no significant difference in SOS between sexes. The SOS of infants showed an inverse correlation with postnatal age, and the decrease of bone SOS with age in premature infants was more marked than in full-term infants. CONCLUSIONS: QUS is suitable for evaluating bone status in infants with high precision. The study offers some basic data for neonates and young infants. PMID- 15889247 TI - Desmoplastic fibroma of the mandible in an infant. AB - We report the CT and MR imaging findings of a case of a desmoplastic fibroma of the mandible in a 6-month-old girl who presented with a rapidly enlarging facial mass and who represents one of the youngest reported children with this rare tumor. Although imaging is non-specific and these lesions are best distinguished histologically, the desmoplastic fibroma should be considered when there is an occurrence of any bubbly or cystic expansile mandibular lesion presenting in childhood. PMID- 15889248 TI - Talar body fatigue stress fractures: three cases observed in elite female gymnasts. AB - OBJECTIVE: To introduce and emphasize the clinical and radiological findings of three talar body fatigue stress fractures in competitive athletes. DESIGN AND PATIENTS: Clinical and radiographic skeletal records of 24,562 athletes taken between 1962 and 2002 were retrospectively reviewed. Among these, 6851 files related to acute foot and ankle injuries or chronic post-traumatic sequelae were found. RESULTS: There were 925 (3.76%) stress fatigue fractures selected from the whole collection. Among these there were three cases (0.32%) of talar body stress fractures diagnosed in elite female gymnasts 15-17 years old. The negative first radiograph become positive 4-6 weeks later. Scintigraphy was positive at an early stage and consistent for the diagnosis. CT and MRI gave positive results 1-2 weeks after the beginning of symptoms which were always greatly diagnostic. CONCLUSIONS: The sports medicine literature lacks reports of talar body fatigue stress fractures. The poor initial sensitivity of radiography makes it problematic to establish an early diagnosis. A wise combination of scintigraphy, CT and MRI has therefore to be relied upon. Familiarity with this rare location for a stress fracture may prevent delayed diagnosis and long-lasting damage, both of which are important factors in competitive athletes. PMID- 15889249 TI - The interplay between innate and adaptive immunity regulates cancer development. AB - There is increasing clinical and experimental evidence that inflammation and cancer are causally linked. Much progress has been made in understanding how inflammatory cells contribute to cancer development; however, it is still largely unknown which molecular mechanisms are responsible for initiation and maintenance of chronic inflammation associated with developing neoplasms. This review will discuss how the adaptive and innate immune systems interact during physiological and chronic inflammation, with a focus on studies revealing new insights into the role of adaptive immune cells as important regulators of chronic inflammation associated carcinogenesis. We will speculate on whether current knowledge about the dysregulated interplay between adaptive and innate immunity during chronic inflammatory disorders might be useful in understanding and targeting the underlying mechanisms of chronic inflammation-associated neoplastic progression. PMID- 15889250 TI - Low-dose IL-2 induces cytokine cascade, eosinophilia, and a transient Th2 shift in melanoma patients. AB - PURPOSE: To assess changes in serum cytokine levels in patients treated concomitantly with or without systemic low-dose IL-2. Vaccination targeted CTL responses to peptide antigens, and IL-2 was coadministered to expand activated CTL. Paradoxically, CTL responses were diminished in patients after 2 weeks of IL 2. We hypothesized that changes in the cytokine milieu may have contributed to this result. EXPERIMENTAL DESIGN: Serum samples were studied from 37 patients enrolled in two clinical trials of a melanoma peptide vaccine administered with or without low-dose IL-2 therapy. Twenty-two patients enrolled in the MEL36 trial received six weekly vaccinations with the four-peptide mixture and were randomized to receive subcutaneous IL-2 (3 x 10(6) IU/m2/day) daily for 6 weeks beginning either at week 1 (upfront group) or at week 4 (delayed group) of vaccine therapy. Fifteen patients on the MEL39 trial were treated with the same vaccine without concurrent IL-2 administration. RESULTS: Circulating levels of IL 5 peaked 1 week after starting IL-2, followed 2 weeks later by a marked eosinophilia, correlating in magnitude with peak IL-5 serum levels. Levels of IFNgamma, GM-CSF, IL-4, IL-10, and IL-12 had no observed relationship to IL-2 administration. At the time of the IL-5 serum peak, PBL responses to mitogen suggested a transient shift to Th2-dominance. CONCLUSIONS: Low-dose IL-2 appears to have induced a transient Th2-dominant secondary cytokine cascade at the time of vaccination, for which eosinophilia is a surrogate marker. For future vaccine therapies targeting cytotoxic T-cell responses, delaying IL-2 until after initiation of immune responses may be more effective. PMID- 15889251 TI - Maturation of circulating dendritic cells and imbalance of T-cell subsets in patients with squamous cell carcinoma of the head and neck. AB - Interactions between dendritic cells (DCs) and T cells play a pivotal role in the regulation and maintenance of immune responses. In cancer patients, various immunological abnormalities have been observed in these immune cells. Here, we investigated proportions and the phenotype of DCs and the cytokine profile of T cell subsets in the peripheral blood of patients with squamous cell carcinoma of the head and neck (SCCHN), using multicolor flow cytometry. The percentage of myeloid (CD11c(+)), but not plasmacytoid (CD123(+)) DCs, was significantly lower (P<0.05) and expression of HLA-DR was significantly decreased in total and myeloid DCs of cancer patients compared to healthy donors. Simultaneous analyses of T-cell subsets in the patients' circulation showed significantly increased proportions of CD4(+) T cells expressing Th1 and Th2 cytokines after ex vivo stimulation without any skewing in the Th1/Th2 ratio. The relative level of HLA DR expression on myeloid or total DCs positively correlated with the Th1/Th2 ratio (P<0.01), and the proportion of total circulating DCs was inversely correlated with that of regulatory CD4(+)CD25+() T cells (P<0.01). These results suggest that the decreased proportion of circulating DCs and decreased HLA-DR expression in DCs may have a major impact on systemic immune responses in patients with SCCHN. PMID- 15889252 TI - Induction of anti B-cell malignance CTL response by subfamily-shared peptides derived from variable domain of immunoglobulin heavy chain. AB - The variable domain of immunoglobulin heavy chain (Ig HV) is well-characterized tumor associated antigen expressed in B-cell malignancies, which may function as a T-cell target. However, T-cell epitopes derived from shared framework regions (FRs) of each IgHV subfamily capable of inducing cytotoxic T lymphocytes (CTLs) against the B-cell malignancy, have not been identified. Using the specific PCR primers of seven IgHV gene subfamilies, we amplified the IgHV gene rearrangement for 108 cases of B-cell acute lymphoblastic leukemia (B-ALL) patients. The IgHV gene rearrangement fragments of B-ALL patients were directly sequenced then classified into seven different subfamilies. The T-cell epitopes encoded by the IgHV gene in the B-ALL patients were predicted by SYFPEITHI and BIMAS programs and compared with those from 56 representative germline IgHV sequences in the genebank. For the HLA-A*0201 locus, we found 1 or 2 top score shared epitopes from each subfamily and got 12 epitopes altogether. Results showed that ten of them were in the FRs. Using an antigen-specific T-cell expansion system, we generated the peptide-special CTLs in vitro, which were capable of killing B lymphoma cell lines that belonged to the same IgHV subfamily in a peptide specific and HLA-restricted manner. Furthermore, we proved that the cytotoxicity of CTLs was IgHV subfamily-specific. These data indicate possible immunotherapy approaches for B-cell malignances patients based on IgHV gene subfamilies. PMID- 15889253 TI - Anti-tumor immune responses following neoadjuvant immunotherapy with a recombinant adenovirus expressing HSP72 to rodent tumors. AB - Gene modification of tumor cells is commonly utilized in various strategies of immunotherapy preventive both as treatment and a means to modify tumor growth. Gene transfer prior to surgery as neoadjuvant therapy has not been studied systematically. We addressed, whether direct intra-tumoral injection of a recombinant adenovirus expressing the immunomodulatory molecule, heat shock protein 72 (ADHSP72), administered prior to surgery could result in sustainable anti-tumor immune responses capable of affecting tumor progression and survival in a number of different murine and rat tumor models. Using intra-dermal murine models of melanoma (B16), colorectal carcinoma (CT26), prostate cancer (TrampC2) and a rat model of glioblastoma (9L), tumors were treated with vehicle or GFP expressing adenovirus (ADGFP) or ADHSP72. Tumors were surgically excised after 72 h. Approximately 25-50% of animals in the ADHSP72 treatment group but not in control groups showed sustained resistance to subsequent tumor challenge. Tumor resistance was associated with development of anti-tumor cellular immune responses. Efficacy of ADHSP72 as neoadjuvant therapy was dependent on the size of the initial tumor with greater likelihood of immune response generation and tumor resistance associated with smaller tumor size at initial treatment. ADHSP72 neoadjuvant therapy resulted in prolonged survival of animals upon re-challenge with autologous tumor cells compared to ADGFP or vehicle control groups. To study the effects on tumor progression of distant metastases, a single tumor focus of animals with multifocal intra-dermal tumors was treated. ADHSP72 diminished progression of the secondary tumor focus and prolonged survival, but only when the secondary tumor focus was <50 mm3 . Our results indicate that gene modification of tumors prior to surgical intervention may be beneficial to prevent recurrence in specific circumstances. PMID- 15889254 TI - CD8 T cells expressing NK associated receptors are increased in melanoma patients and display an effector phenotype. AB - CD8+ T cells can express NK-associated receptors (NKRs) that may regulate their cytolytic function. We have characterized the expression of several NKRs on peripheral blood CD8+ T cells from melanoma patients and compared them to age matched healthy donors. The analysis performed includes HLA class I specific receptors (KIRs, LILRB1 and CD94/NKG2) and other NK receptors like CD57, CD56 and CD16. Melanoma patients showed a higher variability in the expression of NKRs on circulating CD8+ T cells than age-matched healthy donors. NKR expression on CD8+ T cells from melanoma patients showed a significant increase of KIR2DL2/L3/S2 (mAb gl183), CD244, CD57, CD56 and CD16. We have also found an increase of CD8+ CD28- CD27- T cells in melanoma patients. This subset represents terminally differentiated effector cells expressing CD244 and high levels of perforin. The expression of NKRs was also mainly restricted to this T cell subset. Altogether, circulating CD8+ T cells from melanoma patients display a distinct phenotype characterized by downregulation of costimulatory molecules and higher expression of NKRs. We suggest that the increased expression of NKRs on T cells may contribute to the final outcome of the immune response against melanoma both stimulating or inhibiting activation and differentiation to effector cells. Blocking inhibitory receptor function and enhancing activating receptors may represent new strategies with therapeutic potential against melanoma. PMID- 15889255 TI - Tumor counterattack: fact or fiction? AB - Cancer development relies on a variety of mechanisms that facilitate tumor growth despite the presence of a functioning immune system. Understanding these mechanisms may foster novel therapeutic approaches for oncology and organ transplantation. By expression of the apoptosis-inducing protein CD95L (FasL, APO 1L, CD178), tumors may eliminate tumor-infiltrating lymphocytes and suppress anti tumor immune responses, a phenomenon called "tumor counterattack". On the one hand, preliminary evidence of tumor counterattack in human tumors exists, and CD95L expression can prevent T-cell responses in vitro. On the other hand, CD95L expressing tumors are rapidly rejected and induce inflammation in mice. Here, we summarize and discuss the consequences of CD95L expression of tumor cells and its contribution to immune escape. PMID- 15889256 TI - Targeted immunotherapy in acute myeloblastic leukemia: from animals to humans. AB - Immunity against acute myeloid leukemia (AML) is demonstrated in humans by the graft-versus-leukemia effect in allogeneic hematopoietic stem cell transplantation. Specific leukemic antigens have progressively been discovered and circulating specific T lymphocytes against Wilms tumor antigen, proteinase peptide or fusion-proteins produced from aberrant oncogenic chromosomal translocations have been detected in leukemic patients. However, due to the fact that leukemic blasts develop various escape mechanisms, antileukemic specific immunity is not able to control leukemic cell proliferation. The aim of immunotherapy is to overcome tolerance and boost immunity to elicit an efficient immune response against leukemia. We review different immunotherapy strategies tested in preclinical animal models of AML and the human trials that spurred from encouraging results obtained in animal models, demonstrate the feasibility of immunotherapy in AML patients. PMID- 15889257 TI - Complete protection against melanoma in absence of autoimmune depigmentation after rejection of melanoma cells expressing alpha(1,3)galactosyl epitopes. AB - The major barrier for xenotransplantation in humans is the presence of alpha(1-3) Galactosyl epitopes (alphaGal) in xenogeneic tissue and the vast quantities of natural antibodies (Ab) produced by humans against this epitope. The binding of anti-alphaGal Ab to cells expressing alphaGal triggers a complement-mediated hyperacute rejection of target cells. The hyperacute rejection of whole cancer cells, modified to express alphaGal epitopes, could be exploited as a new cancer vaccine to treat human cancers. We tested this hypothesis in alphaGalactosyltransferase knockout (alphaGT KO) mice which, like humans, do not express alphaGal on their cell surfaces and can produce anti-alphaGal Ab. Forty five percent of mice with preexisting anti-alphaGal Ab rejected alphaGal positive melanoma cells (B16alphaGal). These mice remained tumor-free for more than 90 days. The majority of control mice injected with B16Null, alphaGal negative cells succumbed to melanoma. The rejection of B16alphaGal induced strong long-lasting antitumor immunity against B16Null measured by the expansion of cytotoxic T lymphocytes. In addition, mice rejecting B16alphaGal were protected against melanoma since they survived a second rechallenge with B16Null. Protected mice developed antitumor immunity in the absence of autoimmune depigmentation (vitiligo). These results show that rejection of alphaGal positive melanoma cells can efficiently boost the immune response to other tumor associated antigens present in alphaGal negative melanoma cells. This study supports the concept of a novel anticancer vaccine to treat human malignancies. PMID- 15889258 TI - Agrobacterium-mediated transformation as a tool for functional genomics in fungi. AB - In the era of functional genomics, the need for tools to perform large-scale targeted and random mutagenesis is increasing. A potential tool is Agrobacterium mediated fungal transformation. A. tumefaciens is able to transfer a part of its DNA (transferred DNA; T-DNA) to a wide variety of fungi and the number of fungi that can be transformed by Agrobacterium-mediated transformation (AMT) is still increasing. AMT has especially opened the field of molecular genetics for fungi that were difficult to transform with traditional methods or for which the traditional protocols failed to yield stable DNA integration. Because of the simplicity and efficiency of transformation via A. tumefaciens, it is relatively easy to generate a large number of stable transformants. In combination with the finding that the T-DNA integrates randomly and predominantly as a single copy, AMT is well suited to perform insertional mutagenesis in fungi. In addition, in various gene-targeting experiments, high homologous recombination frequencies were obtained, indicating that the T-DNA is also a useful substrate for targeted mutagenesis. In this review, we discuss the potential of the Agrobacterium DNA transfer system to be used as a tool for targeted and random mutagenesis in fungi. PMID- 15889259 TI - [New methods for the prevention of posterior capsule opacification]. AB - Even though tremendous advances have been made especially during the last 10-15 years in terms of surgical techniques and improvement of implant technology, posterior capsule opacification (PCO) still remains a serious long-term complication. New clinical and laboratory studies (especially of autopsy eyes) have improved our understanding of how IOL design and material influence PCO. Sharp edge optic designs of IOLs of various materials have been shown to significantly reduce secondary cataract. The application of pharmacological substances selectively into the capsular bag is now possible due to the development of the PerfectCapsule System for vacuum-sealed capsule irrigation. Major advances in other areas of biotechnology and immunology including gene therapeutic methods offer totally new approaches for the future in the elimination of lens epithelium cells from the capsular bag. This survey gives an update on current and future means and trends to reduce or prevent PCO formation. PMID- 15889260 TI - [Cranial imaging and serology tests are not helpful in tonic pupil. A retrospective study]. AB - BACKGROUND: Tonic pupil (TP) is a common disorder of parasympathetic innervation. In contrast to textbook recommendations, cranial imaging is still being performed in most of the patients with TP. The intention of the present study is to show that cranial imaging is of no benefit. PATIENTS AND METHODS: The medical records of 33 patients with TP were analyzed retrospectively. All patients had undergone a complete ophthalmological, orthoptic, and neurological investigation. Cranial imaging was performed by computed tomography or magnetic resonance imaging. Serology tests were carried out in some of the patients. RESULTS: Diagnostic imaging provided no additional data revealing the underlying cause of TP. CONCLUSIONS: Cranial imaging in isolated tonic pupil is not helpful. Because of therapeutic implications, diagnostic evaluation can be recommended only in patients older than 50 years to exclude giant cell arteritis and syphilis. PMID- 15889261 TI - Selective modulation of task performance by octopamine in honey bee (Apis mellifera) division of labour. AB - Octopamine treatment has previously been shown to increase honey bee foraging behaviour. We determined the effects of octopamine on other tasks to learn how octopamine affects division of labour in honey bee colonies. Octopamine treatment did not increase the rate of corpse removal from the hive, suggesting that elevated brain levels of octopamine do not act to increase the performance of all flight-related tasks. Octopamine treatment also did not increase attendance in the queen's retinue, suggesting that elevated brain levels of octopamine do not act to increase responsiveness to all olfactory stimuli. Consistent with these findings, octopamine treatment enhanced the foraging response to brood pheromone but not the cell capping response, a component of brood care. These results demonstrate a relatively specific form of neuromodulation by octopamine in the regulation of division of labour in honey bee colonies. PMID- 15889262 TI - Unusual features of gall bladder duplication cyst with review of the literature. AB - A 1-month-old female infant presented with a lump in the right hypochondrium extending into the right iliac fossa. Ultrasonography and computed tomography suggested an intestinal duplication cyst or a gall bladder cyst. Exploratory laparotomy revealed it to be a congenital gall bladder duplication cyst without associated complications. It was removed after confirming the presence of a normal gall bladder. The infant's postoperative course was uneventful. This unusual presentation of congenital gall bladder duplication cyst prompted us to report this case. PMID- 15889263 TI - The rare epidemiologic coincidence of diverticular disease and advanced colonic neoplasia. AB - BACKGROUND AND AIMS: In Western industrialised countries the prevalence of neoplastic colonic lesions and diverticular disease markedly increases with age. In contrast, the coincident occurrence of both diseases seems to fall below their individual epidemiologic estimates. Because directly comparing data are rare, this retrospective study evaluates the coincidence of neoplastic lesions and diverticular disease. PATIENT AND METHODS: A total of 1,838 patients from 1986 to 2000 were admitted to the study. For 1,326 patients-56% male (n=741), 44% female (n=585), mean age 64 (+/-11.83 SD)-with a resection due to colonic cancer, the documented findings of colonoscopy, colonic contrast enema, and/or histopathology were analysed with regard to the prevalence of colonic diverticulosis. In 512 patients--51% male (n=263), 49% female (n=249), mean age 60 (+/-12.59 SD)--with a colonic resection due to diverticulitis, the synchronous or metachronous occurrence of neoplastic colonic lesions was recorded using the database of the Tumour Centre, Aachen. To compare the observed results with published epidemiology, statistical analysis included age-referred binomial tests and an age-stratified analysis (Cochran-Mantel-Haenszel test). Odds ratios (OR) were also calculated. P<0.05 was considered to indicate locally statistical significance. RESULTS: In the cancer group, we found a statistically significant reduced rate of diverticula in nearly all age categories and the age-stratified analyses (corresponding OR 0.30-0.51). Consistently, the diverticulitis group revealed a statistically significant decreased rate of advanced colonic neoplastic lesion in nearly all age categories and all age-stratified analyses (corresponding OR 0.13-0.43). CONCLUSION: Our results indicate that patients with colonic neoplastic lesions or diverticular disease probably form heterogeneous groups. Because current results from molecular biology emphasize the impact of the extracellular matrix on the genesis of diverticulosis and colonic cancer, the observed heterogeneity could be an expression of a distinct composition of the local milieu. PMID- 15889264 TI - Narrowband-UVB irradiation decreases the production of pro-inflammatory cytokines by stimulated T cells. AB - Narrow-band ultraviolet B (UVB) phototherapy is an effective treatment for psoriasis. Owing to its limited penetration, the direct effects of UVB are mostly restricted to cells residing in the epidermis and papillary dermis, and are associated with epidermal depletion of Langerhans' cells (LC) and T cells. It has been argued that the depletion of the skin-resident T-cell population may be due to a combination of UVB-induced apoptosis and decreased recruitment from the blood due to lower expression of the required adhesion and homing molecules. We have previously demonstrated that UVB treatment can alter the expression of adhesion molecules by blood lymphocytes, and as these can be influenced by cytokines, the aim of this study was to investigate whether UVB irradiation can also influence the cytokine production of circulating T cells. Four patients with active chronic plaque psoriasis were treated daily with narrow-band 312 nm UVB irradiation and blood samples obtained before treatment and weekly thereafter for 2 weeks. Peripheral blood mononuclear cells (PBMCs) were isolated and cultured with a streptococcal superantigen or a conventional streptococcal antigen preparation, and cell culture supernatants were assayed for various cytokines. When stimulated with the superantigen, PBMCs from UVB-treated psoriasis patients secreted greater amounts of the anti-inflammatory cytokine IL-10, and showed markedly decreased production of IL-1beta, IL-2, IL-5 and IL-6 compared to the pre-treatment values; the production of IFN-gamma, IL-8 and IL-12p70 were also decreased but did not reach statistical significance. Thus, the combination of UVB-induced apoptosis, increased secretion of anti-inflammatory cytokines and decreased trafficking to the skin may help to explain the beneficial effects of UVB treatment on psoriasis and why disease remission can sometimes be sustained for a prolonged period. PMID- 15889265 TI - PTEN-mediated Akt activation in human neocortex during prenatal development. AB - Akt is a crucial factor for cell survival and migration. Phosphatase and tensin (PTEN) negatively regulates cell growth and survival by inhibiting PI3K-dependent signaling. PTEN also blocks Akt phosphorylation, a main downstream molecule of PI3K cascade. So far, no studies have shown PTEN expression and Akt phosphorylation levels in the developing human neocortex. Our hypothesis is that spatial and temporal expression of PTEN is likely to modulate developing human brain cortical modeling by regulating Akt activation. Therefore, our aim is to analyze the expression pattern of PTEN and phospho-Akt levels using immunohistochemistry, Western blot, and semiquantitative analysis in the developing human neocortex (n=13 fetuses from first, second, and third trimesters). PTEN expression was decreased parallel to development, but some cells revealed strong nuclear immunoreactivity in the developing neocortex while the active Akt level was increased. Double immunohistochemistry was performed for proliferating cell nuclear antigen (PCNA)-Tuj1 (as neuronal marker) and PCNA-GFAP (Glial marker) to the subsequent sections of PTEN and Akt-stained slides. PCNA (+) cells were mostly positive for glial fibrillary acidic protein (GFAP) and correlated with active-Akt immunoreactivity. Our results suggest that Akt mediated signaling plays an active role in cell migration, survival, and cerebral cortical modeling throughout prenatal life and that PTEN is the most likely protein to regulate this signaling. PMID- 15889266 TI - Characterisation of disseminated tumor cells in the bone marrow of breast cancer patients by the Thomsen-Friedenreich tumor antigen. AB - The detection of disseminated tumor cells in the bone marrow (DTC-BM) of breast cancer patients has proved prognostic significance in all stages of the disease. Further characterisation of those cells could help to improve the biological understanding of metastases, develop targeted therapies and define surface markers for enrichment techniques. The Thomsen-Friedenreich (TF) antigen has been shown to be a tumor specific antigen in breast cancer. The aim of this study was to investigate the expression of TF on DTC-BM in 25 patients. Bone marrow samples were first double-stained by a Cy3 conjugated cytokeratin (CK) antibody (ab) A45 B/B3 (IgG) and anti-TF ab Nemod 2 (IgM), followed by Cy2 conjugated goat anti mouse IgM ab. For further characterisation samples were also double-stained with anti-TF ab Nemod 2 (IgM), followed by Cy2 conjugated goat anti-mouse IgM ab, and anti MUC1 ab A76-A/C7 IgG, followed by Cy3 conjugated goat anti-mouse IgG. CK positive DTC-BM showed co-expression of TF antigen in 22/23 patients (96%) and 61 of 62 detected cells (98%). Mononuclear BM cells without CK expression were also negative for TF. All of the TF positive cells showed strong MUC1 expression. This is the first study showing co-expression of CK and TF as markers of DTC-BM. Double staining experiments of TF and MUC1 expression showed that MUC1 is the carrier protein of TF in these cells. As TF is a specific marker of DTC-BM, it could be used as a target for antibody based therapy and immunomagnetic enrichment techniques for the isolation of DTC-BM. PMID- 15889267 TI - Belt-like localisation of caveolin in deep caveolae and its re-distribution after cholesterol depletion. AB - Caveolae are specialised vesicular microdomains of the plasma membrane. Using freeze-fracture immunogold labelling and stereoscopic imaging, the distribution of labelled caveolin 1 in caveolae of 3T3-L1 mouse fibroblast cells was shown. Immunogold-labelled caveolin structures surrounded the basolateral region of deeply invaginated caveolae like a belt whereas in the apical region distal to the plasma membrane, the caveolin labelling was nearly absent. Shallow caveolar membranes showed a dispersed caveolin labelling. After membrane cholesterol reduction by methyl-beta-cyclodextrin treatment, a dynamic re-distribution of labelled caveolin 1 and a flattening of caveolar structures was found. The highly curved caveolar membrane got totally flat, and the initial belt-like caveolin labelling disintegrated to a ring-like structure and later to a dispersed order. Intramembrane particle-free domains were still observable after cholesterol depletion and caveolin re-distribution. These results indicate that cholesterol interacting with caveolin structures at the basolateral part of caveolae is necessary for the maintenance of the deeply invaginated caveolar membranes. PMID- 15889268 TI - Quantitative histomorphometric analysis of gonadal steroid receptor distribution in the normal human endometrium through the menstrual cycle. AB - The aim of this study was to test the hypothesis that the distribution of oestrogen receptor beta (ERbeta) and androgen receptor (AR) are related to cell proliferation or correlated with the expression of progesterone receptor (PR) or oestrogen receptor alpha (ERalpha) in the normal human endometrium. Immunohistochemical distribution of immunoreactive ERbeta in well-characterised menstrual cycle biopsy samples was lowest in proliferative endometrial glands, highest in early secretory phase glands and maintained at approximately 20% throughout the rest of the menstrual cycle and was closely correlated with stromal AR and stromal ERbeta expression. Stromal ERbeta was not significantly altered until the menstrual phase of the cycle and was not correlated with the expression of any other antigen in the stroma or endometrial glands except stromal AR. By contrast, glandular AR immunoreactivity was below 5% early in the cycle, increased during the secretory phase and showed strong expression just before menstruation. PR and Ki-67 expression showed strong positive correlations, indicating that PR may be a potent regulator of endometrial proliferation. These data suggest that glandular ERbeta expression is closely associated with a functional secretory role whereas glandular ERalpha and PR are associated with proliferation; glandular AR expression may be the switch required for menstruation. PMID- 15889269 TI - Isolation of adult murine thymic stromal cells that naturally express Notch ligands. AB - This communication summarizes the procedures that enabled isolation of adult murine thymic stromal cell lines that naturally express Notch ligands Jagged-1 or Delta-1. Histochemical characterization of these cell lines, in terms of ligand and cell type, revealed epithelial cells that bear an antigen characteristic of the thymic medulla and express either Jagged-1 or Delta-1. FACS sorting of stromal cells that naturally express these and other ligands is thus feasible, and such cells can be used to investigate the activity of each ligand in Notch mediated commitment to the T-lymphocyte pathway. PMID- 15889270 TI - Analysis of ABCC6 (MRP6) in normal human tissues. AB - To determine the tissue distribution of the ABC transporter ABCC6 in normal human tissues, we analyzed tissue arrays for the presence of ABCC6 mRNA by in situ hybridization and ABCC6 protein by immunohistochemistry using the polyclonal antibody HB-6. We detected ABCC6 mRNA and protein in various epithelial cells of exocrine and endocrine tissues, such as acinar cells in the pancreas, mucosal cells of the intestine and follicular epithelial cells of the thyroid. We obtained a very strong immunostaining for enteroendocrine G cells in the stomach. In addition, ABCC6 mRNA and protein were present in most neurons of the brain, in alveolar macrophages in the lungs and lymphocytes in the lymph node. Immunohistochemisty using the monoclonal antibody M6II-31 confirmed the widespread tissue distribution of ABCC6. The physiological substrate(s) of ABCC6 are yet unknown, but we suggest that ABCC6 fulfills multiple functions in different tissues. The strong immunostaining for ABCC6 in G cells suggests that it plays an important role in these endocrine cells. PMID- 15889271 TI - Characterisation of transverse slice culture preparations of postnatal rat spinal cord: preservation of defined neuronal populations. AB - Spinal cord injury induces degenerative and regenerative processes and complex interactions of neurons with non-neuronal cells. In order to develop an in vitro tool for the investigation of such processes, we prepared and characterised spinal cord slice cultures (SCSC) from Wistar rats (p0-12). SCSC were sustained in vitro up to 12 days and characterised by immunohistochemistry. Calbindin+ neurons, distributed across the entire gray matter, were visible also after longer culture periods. NeuN+ neurons were best preserved in the dorsal horn whereas large NeuN+ and choline acetyltransferase+ motoneurons in the ventral horn vanished after 3 days in vitro. Nestin immunoreactivity was found in animals of all age groups, either in cells interspersed in the ependymal lining around the central canal or in cells resembling protoplasmic astrocytes. Glial fibrillary acidic protein+ astrocytes, initially restricted to the white matter, invaded the gray matter of SCSC early during the culture period. Microglial cells, stained by Griffonia simplicifolia isolectin B4, were rapidly activated in the dorsal tract and in the gray matter but declined in number with time. SCSC derived from p0 or p3 animals showed a better preservation of the cytoarchitecture than cultures derived from older animals. In summary, SCSC undergo degenerative changes, but they contain defined neuronal populations, the cytoarchitecture is partially preserved and the glial reaction is limited. PMID- 15889272 TI - Crosstalk between ABA and auxin signaling pathways in roots of Arabidopsis thaliana (L.) Heynh. AB - Studies of abscisic acid (ABA) and auxin have revealed that these pathways impinge on each other. The Daucus carota (L.) Dc3 promoter: uidA (beta glucuronidase: GUS) chimaeric reporter (ProDc3:GUS) is induced by ABA, osmoticum, and the auxin indole-3-acetic acid (IAA) in vegetative tissues of transgenic Arabidopsis thaliana (L.) Heynh. Here, we describe the root tissue-specific expression of ProDc3:GUS in the ABA-insensitive-2 (abi2-1), auxin-insensitive-1 (aux1), auxin-resistant-4 (axr4), and rooty (rty1) mutants of Arabidopsis in response to ABA, IAA and synthetic auxins naphthalene acetic acid (NAA), and 2, 4 (dichlorophenoxy) acetic acid. Quantitative analysis of ProDc3:GUS expression showed that the abi2-1 mutant had reduced GUS activity in response to ABA, IAA, or 2, 4-D: , but not to NAA. Similarly, chromogenic staining of ProDc3:GUS activity showed that the aux1 and axr4 mutants gave predictable hypomorphic ProDc3:GUS expression phenotypes in roots treated with IAA or 2, 4-D: , but not the diffusible auxin NAA. Likewise the rty mutant, which accumulates auxin, showed elevated ProDc3:GUS expression in the absence or presence of hormones relative to wild type. Interestingly, the aux1 and axr4 mutants showed a hypomorphic effect on ABA-inducible ProDc3:GUS expression, demonstrating that ABA and IAA signaling pathways interact in roots. Possible mechanisms of crosstalk between ABA and auxin signaling are discussed. PMID- 15889273 TI - Motifs specific for the ADR1 NBS-LRR protein family in Arabidopsis are conserved among NBS-LRR sequences from both dicotyledonous and monocotyledonous plants. AB - The activated disease resistance (ADR) 1 gene encodes a protein that possesses an N-terminal coiled-coil (CC) motif, nucleotide-binding site (NBS) and C-terminal leucine-rich repeat (LRR) domains. ADR1 belongs to a small, atypical Arabidopsis thaliana sub-class containing four CC-NBS-LRR genes. The NBS region of most NBS LRR proteins possesses numerous conserved motifs. In contrast, the LRR domain, which is subject to positive selection, is highly variable. Surprisingly, sequence analysis revealed that the LRR domain of the ADR1 sub-class was more conserved than the NBS region. Sequence analysis identified two novel conserved motifs, termed TVS and PKAE, specific for this CC-NBS-LRR sub-class. The TVS motif is adjacent to the P-loop, whereas the PKAE motif corresponded to the inter domain region termed the NBS-LRR linker, which was conserved within the different CC-NBS-LRR classes but varied among classes. These ADR1-specific motifs were employed to identify putative ADR1 homologs in phylogenetically distant and agronomically important plant species. Putative ADR1 homologs were identified in 11 species including rice and in 3 further Poaceae species. The ADR1 sub-class of CC-NBS-LRR proteins is therefore conserved in both monocotyledonous and dicotyledonous plant species. PMID- 15889274 TI - Intermittent claudication due to right common femoral artery compression by a solitary neurofibroma. PMID- 15889275 TI - Low-energy radiofrequency catheter ablation as therapy for supraventricular tachycardia in a premature neonate. AB - A premature neonate with hydrops was born at 32 weeks of gestation after successful direct fetal amiodarone therapy via cordocentesis for incessant supraventricular tachycardia. After birth the tachycardia could not be controlled despite high doses of amiodarone and flecainide and the patient developed severe respiratory and circulatory failure. After 3 weeks, weighing 2 kg, he underwent successful and uncomplicated catheter ablation of a left free-wall accessory pathway using low-energy radiofrequency. CONCLUSION: radiofrequency catheter ablation is rarely used in neonates, but when used with caution may provide the optimal treatment. PMID- 15889276 TI - Fatal coronary artery anomaly presenting as bronchiolitis. AB - During winter outbreaks of respiratory syncytial virus bronchiolitis from 2002 to 2004, three infants presented with a presumptive diagnosis of lower respiratory tract infection and wheezing. The clinical condition in two cases was rapidly progressive and precipitated into intractable shock; clinical and instrumental examinations revealed a cardiac origin of their illness. A subacute presentation permitted a cardiological assessment and a proper treatment in the third infant. An abnormal origin of the left coronary artery from the pulmonary trunk was demonstrated in all cases. The concurrent acute airway infection had a catastrophic effect on the underlying cardiovascular anomaly leading to refractory cardiogenic shock and death. CONCLUSION: Admission chest X-ray film and arterial gas analysis can raise the suspicion of cardiac involvement when treating a severe wheezing episode in young infants. Paediatric cardiological evaluation with two-dimensional echocardiography may eventually reveal this rare condition, whereas cardiac catheterisation with aortography remains the standard means of diagnosis. PMID- 15889277 TI - An unusual combination of EEC syndrome and hypomelanosis Ito due to a p63 mutation. PMID- 15889278 TI - A novel mutation in the PTPN11 gene in a patient with Noonan syndrome and rapidly progressive hypertrophic cardiomyopathy. AB - A male infant with clinical features of Noonan syndrome and rapidly progressive hypertrophic cardiomyopathy is reported. He manifested severe heart failure and failure to thrive. Administration of propranolol and cibenzoline improved ventricular outflow tract obstruction, leading to catch-up growth. Genetic analysis of the patient revealed a novel missense mutation in the PTPN11 gene. CONCLUSION: This is the first description of a patient with a Gln510Glu mutation in the protein-tyrosine phosphatase, non-receptor type 11 gene. This specific mutation may be associated with a rapidly progressive hypertrophic cardiomyopathy. PMID- 15889279 TI - Familial X-linked cardiomyopathy (Danon disease): diagnostic confirmation by mutation analysis of the LAMP2gene. AB - A boy presented at age 2.5 years with mild left ventricular hypertrophy and mild myopathy. Hypertrophic cardiomyopathy progressed relentlessly, leading to death at age 16 years shortly before planned heart transplantation. During the course of the disease, his mother developed severe dilated cardiomyopathy and died of its complications at 46 years of age. The combination of myopathy and cardiomyopathy, the biochemical and electron microscopy findings in a muscle biopsy, and the pedigree suggested Danon disease (MIM 300257), an X-linked lysosomal storage disorder caused by deficiency of lysosome-associated membrane protein-2 (LAMP2). The diagnosis was confirmed by the identification of a novel mutation, G138A, in the LAMP2gene, leading to the premature stop codon W46X. CONCLUSION: Early diagnosis of Danon disease is important for genetic counselling and timely cardiac transplantation, the only effective therapeutic option. PMID- 15889280 TI - Mechanisms of renal damage owing to infection. AB - Urinary tract infection (UTI) is a common bacterial illness in children. It is known to be associated with an increased risk of permanent renal cell damage and scarring which may lead to generation of pathological conditions such as hypertension, pre-eclampsia during pregnancy, renal insufficiency, and end-stage kidney disease. The pathophysiology of renal scarring is still obscure, which makes the prevention of renal damage difficult. During acute infection, there are numerous factors that may contribute to tissue damage. Inflammatory responses are activated by host defense mechanisms as well as by specific bacterial virulence factors. Understanding of these complex mechanisms would be helpful to better identify children at high risk of developing renal scarring following UTI. PMID- 15889281 TI - Can azathioprine and steroids alter the progression of severe Henoch-Schonlein nephritis in children? AB - To evaluate the effect of azathioprine with steroids on the clinical course and histologic parameters of severe Henoch-Schonlein nephritis (HSN), 20 patients with a median age of 9.3 years (range 4.4-17 years) and a follow-up period of 4.8 years (range 1-14 years) were included in this study. The patients were divided into two groups. Ten patients received azathioprine with steroids for 8 months (group A), and the other ten received steroids alone (group B). All patients underwent renal biopsy at presentation, and ten of them who were treated with azathioprine underwent follow-up biopsy after therapy. Six patients in group A achieved clinical remission, and the remaining four showed minor urinary abnormalities at the latest observation. The histological grades of the International Study of Kidney Disease in Children (ISKDC) improved in four of the ten patients, and the activity index decreased significantly from 6.8+/-2.0 to 4.3+/-2.3 (p=0.016). In group B, however, four patients had normal urine and renal function; two had minor urinary abnormalities; one had active renal disease, and three had chronic renal insufficiency, at the latest observation. The combination treatment of azathioprine and steroids may be beneficial in ameliorating histopathological features and improving the clinical course of severe HSN. PMID- 15889282 TI - Complete remission of post-transplant FSGS recurrence by long-term plasmapheresis. AB - Focal segmental glomerulosclerosis (FSGS) is known to recur in approximately 30% of renal allografts with graft loss in about half of these cases. The exact etiology remains unclear, though a putative circulating permeability factor or loss of inhibitory substances is being discussed. Different therapeutic approaches have been used. We report on a 10-year-old Arabian boy with a recurrence of FSGS immediately after transplantation. In addition to intensifying immunosuppressive therapy with high-dose cyclosporin A and cyclophosphamide, plasmapheresis was initiated and remission was achieved after 8 months. Three weeks after cessation of plasmapheresis a relapse occurred. Plasmapheresis was resumed and remission was achieved again after four additional sessions. The interval between plasmapheresis treatments was then gradually increased and fourteen months after transplantation plasmapheresis was stopped again. Since then (1.5 years after cessation of treatment) the patient has been in complete remission without any further episode of proteinuria. In conclusion, complete and sustained remission with stable renal function was achieved in our patient by long-term plasmapheresis in combination with intensified immunosuppression. Therefore, continuation of plasmapheresis treatment should be considered even in the situation of initial non-response. PMID- 15889285 TI - Post-fire, seasonal and annual dynamics of the ectomycorrhizal community in a Quercus ilex L. forest over a 3-year period. AB - Two study plots, burned and control, were established in autumn 1998 in a Quercus ilex forest located in northern Spain, part of which had been affected by a low intensity fire in 1994. Soil samples for ectomycorrhizae (ECM) were taken over a 3-year period in each study plot in spring, summer, autumn and winter. ECM morphotypes were identified and the relative abundance of each morphotype in each soil sample calculated, along with species richness, Shannon diversity index and percentage of mycorrhization in each soil sample. The relative abundance of certain ECM morphotypes differed between burned and control plots, and the percentage of mycorrhizal tips was significantly lower in the burned than in the control plot. Nevertheless, there were no significant differences in the diversity, species richness or species composition of the ECM community in the burned and control plots. The dominant ECM morphotypes in both stands were Cenococcum geophilum and several thelephoroid fungi. Sphaerosporella brunnea and Pisolithus tinctorius thrived especially in the burned plot, whereas three ectomycorrhizal morphotypes assigned to the genus Hebeloma were especially abundant in the control plot. There was no significant variation in the relative abundance of the ECM morphotypes between seasons, but ECM community species richness was highest in autumn and lowest in summer. The percentage of mycorrhizal tips reached a maximum in winter, with its minimum in autumn. Collection of samples over the 3-year period also enabled us to detect a significant increase in percentage of ECM colonisation in the burned stand over time. PMID- 15889286 TI - EPR analysis of multiple forms of [4Fe-4S](3+) clusters in HiPIPs. AB - The electron paramagnetic resonance (EPR) spectrum from the [4Fe-4S](3+) cluster in several high-potential iron-sulfur proteins (HiPIPs) is complex: it is not the pattern of a single, isolated S=1/2 system. Multifrequency EPR from 9 to 130 GHz reveals that the apparent peak positions (g values) are frequency-independent: the spectrum is dominated by the Zeeman interaction plus g-strain broadening. The spectra taken at frequencies above the X-band are increasingly sensitive to rapid passage effects; therefore, the X-band data, which are slightly additionally broadened by dipolar interaction, were used for quantitative spectral analysis. For a single geometrical [4Fe-4S](3+) structure the (Fe-Fe)(5+) mixed-valence dimer can be assigned in six different ways to a pair of iron ions, and this defines six valence isomers. Systematic multicomponent g-strain simulation shows that the [4Fe-4S](3+) paramagnets in seven HiPIPs from different bacteria each consist of three to four discernible species, and these are assigned to valence isomers of the clusters. This interpretation builds on previous EPR analyzes of [4Fe-4S](3+) model compounds, and it constitutes a high-resolution extension of the current literature model, proposed from paramagnetic NMR studies. PMID- 15889287 TI - A hypothesis for GPCR activation. AB - Growing evidence that rhodopsin (RD) and related G protein-coupled receptors form functional dimers/oligomers, followed by direct proof (using atomic force microscopy) that in the retina disc membrane RD associates into a paracrystalline network of rows of dimers, need models of the RD-transducin (Gt) complex that would envision an optimal RD dimer/oligomer able to satisfy all well-documented interactions with Gt. Of the models proposed so far, only a few refer to RD dimers and only one of them proposes a complex of Gt with an RD oligomer (Filipek S, Krzysko KA, Fotiadis D, Liang Y, Saperstein DA, Engel, A, Palczewski K Photochem Photobiol Sci 3: 628-638, 2004). This paper puts forward a hypothesis on another arrangement of RD monomers into the reported network of rows of dimers. Arguments for the compatibility of this set-up with interactions and activation of RD in the complex with Gt, in particular, with the well-documented movement of transmembrane helix 6 and cytosolic loop 3, which is vital for RD activation, are provided and discussed. PMID- 15889288 TI - On the nature of bonding in HCOOH...Ar and HCOOH...Kr complexes. AB - The chemical interaction in HCOOH...Ng (Ng=Ar, Kr) complex was analyzed by topological analysis of the electron density based on Atoms-In-Molecules theory. For all computationally stable equilibrium structures of 1:1 HCOOH...Ng complexes, an intermolecular bond path with a "bond" critical point was found and perturbation of formic acid (FA) atomic basins and electron density was observed. The intermolecular interaction between the two complex subunits can be classified, according to its nature, as a closed-shell van der Waals type of interaction. However, one of the computed structures (complex II), pictures a noble gas atom attached linearly to the acidic O-H tail of FA. In this particular case, the electron density at the intermolecular bond critical point was found to resemble a hydrogen-bonded system and thus, may be termed a hydrogen-bond-like interaction. This change in the nature of the interaction is also shown by large perturbations of the FA properties found for this complex structure. The structural and vibrational perturbations are larger than for the other three structures and they increase for the Kr complexes compared to the Ar complex. [Figure: see text]. Electron density analysis of HCOOH...Ng (Ng=Ar,Kr) complex. PMID- 15889289 TI - Modeling of adsorption in nanopores. AB - Adsorption in nonporous materials has been studied using Grand Canonical Monte Carlo simulations. We discuss three types of materials: (a) a model of cylindrical pores with smooth walls, representing MCM-41 like materials, (b) a model of cylindrical pores with regular structured walls (model of carbon nanotubes) and (c) a material with crystalline wall structure (zeolites). Typical problems related to the stability of adsorbed layers have been analyzed. We have shown that the mechanism of adsorption is strongly dependent on the structure of the pore walls. In the case of amorphous walls it may lead to metastable configurations. In nanotubes, the ordered corrugation structure of walls determines the low temperature structure of the adsorbed system. In 3D ordered porous system, such as zeolites, the mechanism of adsorption is mostly determined by characteristic sites of adsorption. [Figure: see text]. Adsorbed atoms and energy fluctuations at the pressure of the first layer formation of krypton atoms: (a) instantaneous numbers of adsorbed atoms (per nm2 of the pore wall) as a function of the time of simulation (Monte Carlo steps) observed in a relatively long run, (b) the bimodal distribution of the energy fluctuations is a consequence of the behavior of the systems as shown in (a). PMID- 15889290 TI - An unusual feature of end-substituted model carbon (6,0) nanotubes. AB - We have examined the effects of substituents on the computed electrostatic potentials V(S)(r) and average local ionization energies I(S)(r) on the surfaces of model carbon nanotubes of the types (5,5), (6,1) and (6,0). For the (5,5) and the (6,1), the effects upon both V(S)(r) and I(S)(r) of substituting a hydroxyl group at one end are primarily localized to that part of the system. For the (6,0) tube, however, a remarkable change is observed over its entire length, with V(S)(r) showing a marked gradation from strongly positive at the substituted end to strongly negative at the other; I(S)(r) correspondingly goes from higher to lower values. Replacing OH by another resonance- donor, NH2, produces similar results in the (6,0) system, while the resonance withdrawing NO2 does the opposite, but in equally striking fashion. We explain these observations by noting that the arrangement of the C-C bonds in the (6,0) tube facilitates charge delocalization over the full length and entire surface of the tube. Substituting NH2 and NO2 at opposite ends of the (6,0) tube greatly strengthens the gradations in both V(S)(r) and I(S)(r). The first hyperpolarizability of this system was found to be nine times that of para-nitroaniline, suggesting possible nonlinear optical applications. [figure: see text]. HF/STO-5G electrostatic potential on outer surface of open (6,0) C72H10NH2NO2. The nitro group is at the right end of the tube, the amino group at the left. In eV: purple is less than 14, blue is between 14 and 15, green is between 15 and 16.5, yellow is between 16.5 and 17.5, and red is more than 17.5. PMID- 15889291 TI - Evaluating molecular similarity using reduced representations of the electron density. AB - A model system of four benzodiazepine-like ligands for the central benzodiazepine receptors (CBRs) and peripheral benzodiazepine receptors (PBRs)is examined using a genetic algorithm procedure (GAGS) designed for evaluating molecular similarity. The method is based on the alignment of reduced representations generated from the critical points of the electron density computed at medium crystallographic resolution. The results are further characterized by a comparison with alignments produced by MIMIC, a field-based superimposition method that matches both steric and electrostatic molecular fields. The alignments produced by the two methods are generally seen to be consistent. The relationships of the compounds' binding affinities for both CBRs and PBRs to the alignments determined by GAGS yield a set of structural features required for significant binding to benzodiazepine receptors. Benefits of using reduced representations for evaluating molecular similarities and for constructing pharmacophore models are discussed. PMID- 15889292 TI - Effect of Pt and Sn on the adsorption of n-heptane in gamma-Al2O3 catalyst models. AB - The Grand Canonical Monte-Carlo (GCMC) method has been used to carry out simulations of the adsorption of n-heptane in models of naphtha-reforming catalysts. Models used in the study differed in the number and distribution of metal atoms-Pt and Sn. The number of adsorbed n-heptane molecules grows linearly with increasing number of metal atoms. The effect of Pt content on the adsorption of n-heptane molecules is most distinct at approximately 100 kPa and within the lower range of the temperatures investigated. In the models of bimetallic catalysts, the effect of the two metals is additive. [Figure: see text]. Effect of Pt and Sn on number of n-heptane molecules adsorbed in Al2O3 catalyst in 773 K and 1000 kPa. PMID- 15889293 TI - A new visualization scheme of chemical energy density and bonds in molecules. AB - Covalent bond describes electron pairing in between a pair of atoms and molecules. The space is partitioned in mutually disjoint regions by using a new concept of the electronic drop region R(D), atmosphere region R(A), and the interface S (Tachibana in J Chem Phys 115:3497-3518, 2001). The covalent bond formation is then characterized by a new concept of the spindle structure. The spindle structure is a geometrical object of a region where principal electronic stress is positive along a line of principal axis of the electronic stress that connects a pair of the R(D)s of atoms and molecules. A new energy density partitioning scheme is obtained using the Rigged quantum electrodynamics (QED). The spindle structure of the stress tensor of chemical bond has been disclosed in the course of the covalent bond formation. The chemical energy density visualization scheme is applied to demonstrate the spindle structures of chemical bonds in H2, C2H6, C2H4 and C2H2 systems. [Figure: see text]. Field theory of the energy density. PMID- 15889294 TI - Structural and functional characterization of AtPTR3, a stress-induced peptide transporter of Arabidopsis. AB - A T-DNA tagged mutant line of Arabidopsis thaliana, produced with a promoter trap vector carrying a promoterless gus (uidA) as a reporter gene, showed GUS induction in response to mechanical wounding. Cloning of the chromosomal DNA flanking the T-DNA revealed that the insert had caused a knockout mutation in a PTR-type peptide transporter gene named At5g46050 in GenBank, here renamed AtPTR3. The gene and the deduced protein were characterized by molecular modelling and bioinformatics. Molecular modelling of the protein with fold recognition identified 12 transmembrane spanning regions and a large loop between the sixth and seventh helices. The structure of AtPTR3 resembled the other PTR type transporters of plants and transporters in the major facilitator superfamily. Computer analysis of the AtPTR3 promoter suggested its expression in roots, leaves and seeds, complex hormonal regulation and induction by abiotic and biotic stresses. The computer-based hypotheses were tested experimentally by exposing the mutant plants to amino acids and several stress treatments. The AtPTR3 gene was induced by the amino acids histidine, leucine and phenylalanine in cotyledons and lower leaves, whereas a strong induction was obtained in the whole plant upon exposure to salt. Furthermore, the germination frequency of the mutant line was reduced on salt-containing media, suggesting that the AtPTR3 protein is involved in stress tolerance in seeds during germination. PMID- 15889295 TI - Superconductivity: small steps towards the "grand unification". AB - The existence of various families of super conducting materials and their T(C) values are qualitatively rationalized within a simple model. Novel families of superconducting materials, particularly those based on fluoride and hydride anions, are predicted. [Figure: see text]. We predict that existing families of moderate- and high-T(C) superconductors should hopefully be enriched by novel compounds containing hardly polarizable anions (such as F(-)). Covalent chlorides and hydrides also merit careful exploration. PMID- 15889296 TI - From cluster calculations to molecular materials: a mixed pseudopotential approach to modeling mixed-valence systems. AB - In this paper we present a technique for finding an appropriate parameterization of ultrasoft pseudopotentials for modeling mixed-valence materials. For the example of hexacyanometallate molecular building blocks, we show how ionic cluster calculations can be used to determine a set of parameters for the metal centers. Pseudopotentials chosen in such a way are then shown to be suitable for periodic calculations of the corresponding mixed-valence materials (e.g., Prussian Blue). PMID- 15889297 TI - Towards a tunable molecular memory that fits into a (10 A)3 cube. AB - Using DFT calculations we predict a novel family of small amino acid-based memory exhibiting molecules, namely complexes of triply deprotonated cysteine (or its derivatives) with selected hexavalent transition metals. We show that the energy difference of the 0 and 1 electromers--corresponding to two chemically distinct oxidation states of a metal, M(VI) and M(IV)--can be tuned towards the thermodynamic equilibrium via deliberate chemical substitutions in the cysteine ligand. Although the thermal stability of both electromers is computed to be rather small at ambient temperatures (due to facile interchange of states 0 and 1 via a low-lying M(V) configuration), we hope to improve stability in related systems. [Figure: see text]. We propose novel kind of tunable molecular memory, based on intramolecular 2e- redox reaction. PMID- 15889298 TI - A case of Aspergillus mediastinitis after heart transplantation successfully treated with liposomal amphotericin B, caspofungin and voriconazole. AB - Reported here is a case of mediastinitis caused by Aspergillus fumigatus and Staphylococcus epidermidis following a heart transplantation that was successfully treated with amphotericin B in combination with new antifungal drugs (caspofungin and voriconazole), antibiotics and superficial wound drainage. A review of the literature revealed that Aspergillus as a cause of mediastinitis has been rarely described. In the few existing reports, evolution was generally fatal, especially in immunocompromised patients, despite treatment with antifungal drugs and surgery. PMID- 15889299 TI - Successful treatment of a disseminated Nocardia brasiliensis infection. PMID- 15889300 TI - Evaluation of pharmacokinetic interaction between cetirizine and ritonavir, an HIV-1 protease inhibitor, in healthy male volunteers. AB - BACKGROUND: Serious adverse effects have been observed with some non-sedative H1 antihistamines (terfenadine and astemizole) when they were associated with drugs known to inhibit their metabolism. However, this is not a class effect, and this interaction should be considered on a case-by-case basis. The aim of this study was to evaluate the potential of pharmacokinetic interaction between cetirizine and ritonavir, the most potent cytochrome P450 (CYP) inhibitor. METHODS: An open label, single-center, one-sequence crossover pharmacokinetic study was conducted in three running periods: cetirizine (CTZ) alone, ritonavir (RTV) alone and then CTZ plus RTV. For each period, steady-state pharmacokinetics were obtained. RTV and CTZ plasma concentrations were determined using validated liquid chromatography methods. The statistical method was based on a 90% confidence interval (CI) for the ratio of population geometric means (combination/drug alone) for each drug and for each parameter [area under the plasma concentration versus time curve (AUC(0-tau,ss)), value of maximum plasma concentration (C(max,ss))] and compared to bioequivalence ranges 80-125% and 70-143% for AUC(0 tau,ss) and C(max,ss), respectively. RESULTS: Among the 17 male subjects enrolled (26.4 +/- 8.6 years), 16 completed the study (1 withdrawal after the first period). The RTV pharmacokinetic parameter values were not affected by CTZ co treatment. With RTV, a 42% increase in the CTZ AUC(0-tau,ss) (3406 versus 4840 microgh/l, 90% CI of 128-158%), a 53% increase in the CTZ elimination half-life (7.8 h versus 11.9 h, P = 0.001), a slight increase (15%) in the CTZ apparent volume of distribution (V(d,ss)/f) (34.7 l versus 39.8 l, P = 0.035), a 29% decrease in the CTZ apparent total body clearance (49.9 ml/min versus 35.3 ml/min, P < 0.001) and bioequivalent C(max,ss) (374 microg/l versus 408 microg/l) were observed. No serious drug related adverse effects were notified. CONCLUSIONS: CTZ does not significantly affect the pharmacokinetic parameters of RTV, and the association does not, thus, require a modification of the dosage of the protease inhibitor. The increased extent of exposure to CTZ in healthy subjects, in the presence of RTV administered at high doses, remained in the same range as previously observed in the elderly or in mildly renally impaired subjects. PMID- 15889301 TI - Prolactinergic and dopaminergic mechanisms underlying sexual arousal and orgasm in humans. AB - Dopaminergic mechanisms play a major role in modulating sexual behavior in humans and animals. Animal data demonstrate important interactions between the dopaminergic and prolactinergic system. As recently demonstrated, dopamine agonists have facilitatory properties for penile erection but may also enhance sexual drive and orgasmic quality. In contrast, chronic elevations of prolactin inhibit appetitive as well as consummatory parameters of sexual behavior. Recent human studies show a marked increase in prolactin after orgasm in males and females. Concerning the biological relevance of acute prolactin alterations after orgasm, prolactin might serve as a neuroendocrine reproductive reflex for peripheral reproductive organs. Alternatively, prolactin may feedback to dopaminergic neurons in the central nervous system and thereby modulate sexual drive and satiation. Here, we provide a brief overview of the physiology of dopamine and prolactin in regulating sexual behavior. In addition, recent experimental and clinical evidence for a postulated feedback mechanism for prolactin and its implications for orgasmic disorders are discussed. PMID- 15889302 TI - Methyltetrahydrophthalic acid in urine as an indicator of occupational exposure to methyltetrahydrophthalic anhydride. AB - OBJECTIVE: To investigate whether methyltetrahydrophthalic acid (MTHP acid) in urine can be used as a biomarker for exposure to methyltetrahydrophthalic anhydride (MTHPA). METHODS: Workers occupationally exposed to MTHPA were studied in combination with one of the authors, who was experimentally exposed to MTHPA. Air levels of MTHPA were determined by personal sampling in the breathing zone. The MTHPA in air was sampled by silica gel and analyzed by gas chromatography (GC) with electron-capture detection. Urinary levels of MTHP acid, a metabolite of MTHPA, were determined in 15 subjects in total. Urine was collected from 14 workers immediately before the start of the work shift and then after 4 and 8 h, and from one of the authors at intervals during 24 h. MTHP acid in urine was analyzed by GC with mass spectrometric detection. RESULTS: The time-weighted average (TWA) air levels ranged from 1.0 microg to 200 microg MTHPA/m3 during 8 h work shifts. The urinary levels of MTHP acid increased during exposure and decayed after the end of exposure, with an estimated half-time of about 3 h. A close correlation was found between the TWA air levels of MTHPA and creatinine adjusted MTHP acid levels in urine collected at the end of the shift (r = 0.955; P < 0.0001). The current occupational exposure limit of 50 microg MTHPA/m3 (Japan Society for Occupational Health) corresponded to about 1300 microg MTHP acid/g creatinine, which was equivalent to about 900 nmol/mmol creatinine in the International System of Units (SI). CONCLUSIONS: These results indicate that the determination of MTHP acid in urine is suitable for use in the biological monitoring of MTHPA exposure. PMID- 15889303 TI - Osteoprotegerin and the receptor activator of NF-kappa B ligand in the serum and synovial fluid. A comparison of patients with longstanding rheumatoid arthritis and osteoarthritis. AB - We examined OPG and soluble RANKL in the serum (sOPG, sRANKL) and synovial fluid (synOPG, synRANKL) in patients with rheumatoid arthritis (RA) and osteoarthritis (OA). OPG and RANKL were measured in 85 patients (44 with RA, 41 patients with OA) in serum and synovial fluid as well. For measuring of OPG and RANKL ELISA tests were used. The results of OPG and RANKL were compared with clinical and radiological scores. We found a negative correlation for OPG and RANKL in synovial fluids: not only for the whole group of patients (P < 0.003, r = -0.32), but also for the subgroups (RA: P < 0.04, r = -0.28, OA: P < 0.002, r = -0.54). SRANKL and synRANKL were positively correlated in the whole group (P < 0.01, r = 0.25) and in the OA group (P < 0.02, r = 0.35); the RA group was showing a trend (P < 0.063, r = 0.24), however. Serum OPG was lower in RA, synOPG higher in OA. The difference between the two patient groups was only significant for synOPG (P < 0.03, r = 0.056), but not for sOPG (P < 0.09, r = 0.19), sRANKL (P < 0.43, r = 0.85) or synRANKL (P < 0.11, r = 0.22). The synOPG:synRANKL ratio was significantly correlated with the Larsen score (P < 0.004, r = 0.38). Synovial OPG is significantly decreased in rheumatoid joints, whereby synovial RANKL is increased. Lower synOPG could reflect a lower protective effect on bone, thus leading to an earlier and more pronounced bone destruction in RA. However, the effect of different mediators for joint destruction in RA and OA seems not to be important to the pathophysiological changes in the joints. The upregulation of serum OPG might be the result of the inflammation; in contrast, an upregulation of RANKL could not be found in the serum of patients with RA and OA. PMID- 15889304 TI - The use of PRIME-MD questionnaire in a rheumatology clinic. AB - OBJECTIVE: To determine whether the Primary Care Evaluation of Mental Disorders 1 page Brief Patient Health Questionnaire (PRIME-MD 1-page PHQ) can serve as: (1) a diagnostic test for fibromyalgia syndrome (FM), or (2) a questionnaire through which internists can be alerted to otherwise hidden mental disorders in patients attending internal medicine clinics. METHOD: Two hundred and thirteen consecutive patients attending a rheumatology clinic were given the PRIME-MD 1-page PHQ and seen by a rheumatologist who was blind to the PRIME-MD diagnosis. RESULTS: The PRIME-MD 1-page PHQ pointed to Major Depressive Disorder in 33.3% of FM patients, Other Depressive Disorder in 33.3% of FM patients, and Panic Disorder in 22.2% of FM patients (all of whom also had Major Depressive Disorder), as compared to 13.1, 13.1, and 3.0% respectively in patients with other rheumatic disorders. However, when used as a diagnostic test for FM, the PRIME-MD 1-page PHQ did not have adequate diagnostic value. When all the PRIME-MD 1-page PHQ diagnoses were compiled, however, a trend was observed. Compared to the rates of mental disorders in both the normal population and in primary care practices, the rates found in this rheumatology clinic were higher. CONCLUSIONS: The PRIME-MD 1-page PHQ is not an adequate diagnostic test for FM. Because FM is primarily a somatization disorder that draws its symptoms from other current diseases, it may in fact be impossible to diagnose FM based on specific symptoms alone. However, the PRIME-MD 1-page PHQ proved to be a useful diagnostic tool in a rheumatology clinic. It helped to alert the physician to the possibility of an elevated frequency of mental disorders that would otherwise have gone unnoticed and untreated. PMID- 15889305 TI - Clinical quiz: a pediatric case presenting with fever and diffuse myalgia. AB - Familial Mediterranean fever (FMF) is a multisystem disease characterized by recurrent polyserositis episodes seen in certain ethnic groups. In recent years the clinical picture of FMF has been expanded and severe myalgia is a frequently recognized component of the syndrome. Protracted febrile myalgia syndrome (PFMS), characterized by severe paralyzing myalgia, high fever, abdominal pain, diarrhea, arthritis/arthralgia, and transient vasculitic rashes mimicking Henoch-Schonlein purpura, was first described in patients with FMF in 1994. We describe an 11-year old Turkish girl with a second attack of PFMS before being diagnosed as having FMF, emphasizing the importance of myalgia for the diagnosis of FMF even in the absence of other symptoms. PMID- 15889306 TI - Knee arthritis as a rare inaugural manifestation of adult leukemia. PMID- 15889307 TI - Polycystins: polymodal receptor/ion-channel cellular sensors. AB - Transient receptor potential (TRP) channel proteins are divided into seven subgroups that are currently designated as TRPC (canonical), TRPV (vanilloid), TRPM (melastatin), TRPN (NOMP-C, from no mechanoreceptor potential-C), TRPA (ankyrin-like with transmembrane domains 1) and TRPP (polycystin). TRPC, TRPV and TRPM are related to canonical TRP proteins whereas TRPN, TRPA and TRPP (polycystin) are more divergent. Most TRP channels are linked to sensory stimuli, including phototransduction, thermosensation and mechanosensation. The TRPP subfamily was named after its founding member, polycystin kidney disease-2 (PKD2), a gene product mutated in many cases of autosomal dominant polycystic kidney disease (ADPKD). ADPKD is a major inherited nephropathy, affecting over 1:1,000 of the worldwide population, characterized by the progressive development of fluid-filled cysts from the tubules and collecting ducts of affected kidneys. Loss-of-function mutations in either polycystin-2, a non-selective cation channel, or polycystin-1 (PKD1), a large plasma membrane integral protein, give rise to ADPKD. PKD1 and PKD2 are thought to function together as part of a multiprotein receptor/ion-channel complex or independently and may be involved in transducing Ca(2+)-dependent mechanosensitive signals in response to cilia bending in renal epithelial cells and endodermally derived cells. Further information on the growing number and physiological properties of these TRP polycystins is the basis of this review. PMID- 15889308 TI - The rat Na+-sulfate cotransporter rNaS2: functional characterization, tissue distribution, and gene (slc13a4) structure. AB - Inorganic sulfate is essential for numerous functions in mammalian physiology. In the present study, we characterized the functional properties of the rat Na+ sulfate cotransporter NaS2 (rNaS2), determined its tissue distribution, and identified its gene (slc13a4) structure. Expression of rNaS2 protein in Xenopus oocytes led to a Na+-dependent transport of sulfate that was inhibited by phosphate, thiosulfate, tungstate, selenate, oxalate, and molybdate, but not by citrate, succinate, or DIDS. Transport kinetics of rNaS2 determined a K(M) for sulfate of 1.26 mM. Na+ kinetics determined a Hill coefficient of n=3.0+/-0.7, suggesting a Na+:SO4 (2-) stoichiometry of 3:1. rNaS2 mRNA was highly expressed in placenta, with lower levels found in the brain and liver. slc13a4 maps to rat chromosome 4 and contains 17 exons, spanning over 46 kb in length. This gene produces two alternatively spliced transcripts, of which the transcript lacking exon 2 is the most abundant form. Its 5' flanking region contains CAAT- and GC box motifs and a number of putative transcription factor binding sites, including GATA-1, SP1, and AP-2 consensus sequences. This is the first study to characterize rNaS2 transport kinetics, define its tissue distribution, and resolve its gene (slc13a4) structure and 5' flanking region. PMID- 15889309 TI - Post-traumatic early epilepsy in pediatric age group with emphasis on influential factors. AB - OBJECTIVE: Posttraumatic epilepsy in the pediatric age group is mostly seen within the first week. An acute posttraumatic epileptic fit, which may induce secondary insults, should be hindered. The aim of the study is to define the risk factors for posttraumatic early epilepsy (PTEE) and the indications for prophylactic therapy. METHODS: In this survey, a total of 1,785 pediatric patients--under the age of 16--are studied. The majority of the patients (1,655) were treated in Haydarpasa Numune Hospital within the years 1993-1999. The rest, which consists of 130 patients, were treated in Inonu University Turgut Ozal Medical Center between the years 2001 and 2003. The patients were categorized according to age, gender, neurological manifestations, type of trauma, cranial pathology, number and type of epileptic fits, the interval between trauma and convulsion, electroencephalogram findings, and antiepileptic therapy. All these factors were challenged due to their effect on the evolution of PTEE. RESULTS: Only 149 cases had PTEE (8.4%). There was no correlation between gender and the incidence of PTEE. The data showed that 11.7% of the patients at or under the age of 3 (p=0.00072), 30.8% of the patients with severe head injury (Glasgow Coma Scale=3-8; Children's Coma Scale = 3-8; p=0.00000), 19.3% of the patients with depressed skull fractures (p=0.00038), 13.7% of the patients with intraparenchymal hemorrhage (p=0.0000072), and 21.6% of the patients with cerebral edema (p=0.000008) had PTEE. Only 20% of the patients with PTEE had a Glasgow Outcome Scale (GOS) of 3 or less (p=0.0000075). CONCLUSION: Those patients at or under the age of 3, with severe head injury, cerebral edema, intraparenchymal hemorrhage, or depressed skull fracture, have a higher incidence of PTEE. Moreover, because the GOS of these patients are prone to be worse, antiepileptic therapy in acute stage may be effective in preventing the secondary brain damage. PMID- 15889310 TI - Functional capacity and body mass index in patients with sellar masses--cross sectional study on 403 patients diagnosed during childhood and adolescence. AB - RATIONALE: We analyzed the impact of tumour localization and histology on functional capacity (FC) and body mass index (BMI) in children with sellar masses. METHODS: FC was evaluated using the ability scale Fertigkeitenskala Munster-Heidelberg in 403 children and adolescents with sellar masses (276 craniopharyngioma, 14 germinoma, 21 optic/chiasmatic glioma, 40 hypothalamic glioma, 13 cysts of Rathke's cleft and 39 other sellar masses). Besides tumour localization, the influence of gender, irradiation and age at diagnosis and at evaluation on FC and BMI was analyzed. General linear models with explanatory influential variables were built. RESULTS: In multivariate analysis, only age at diagnosis (p<0.001) and hypothalamic involvement (p=0.005) had relevant impact on FC. The second model showed BMI at diagnosis (p<0,001), hypothalamic involvement (p<0.001) and craniopharyngioma (p=0,004) to influence BMI at the latest evaluation. CONCLUSION: We conclude that hypothalamic involvement and young age at diagnosis had major impact on FC and BMI and should be considered as risk factors for impaired rehabilitation. PMID- 15889311 TI - New developments in the understanding of the cation diffusion facilitator family. AB - Cation diffusion facilitator (CDF) proteins are a phylogenetically ubiquitous family of intermembrane transporters generally believed to play a role in the homeostasis of a wide range divalent metal cations. CDFs are found in a host of membranes, including the bacterial cell membrane, the vacuolar membrane of both plants and yeast, and the golgi apparatus of animals. As such, they are potentially useful in the engineering of hyperaccumulative phytoremediation systems. While not yet sufficient for reliable biotechnological manipulation, characterization of this family is proceeding briskly. Experimental data suggests that CDFs are generally homodimers that use proton antiport to drive substrate translocation across a membrane. This translocation of both substrate and protons is likely mediated by a combination of histidines, aspartates, and glutamates. Functional data has suggested that CDFs are not limited to metal homeostasis roles, as some appear to be determinants in the operation of high-volume metal resistance systems, and others may facilitate cation-donation as a means of signal transduction. This review seeks to give an overview of the data prompting these conclusions, while presenting additional data whose interpretation is still contentious. PMID- 15889312 TI - Effect of impact exercise on bone mineral density in elderly women with low BMD: a population-based randomized controlled 30-month intervention. AB - Evidence of the effect of exercise on bone loss comes mainly from studies in voluntary postmenopausal women, and no population-based, long-term interventions have been performed. The purpose of this population-based, randomized, controlled trial was to determine the effect of long-term impact exercise on bone mass at various skeletal sites in elderly women with low bone mineral density (BMD) at the radius and hip. Participants (n=160) were randomly assigned to 30 months either of supervised and home-based impact exercise training or of no intervention. The primary outcome measures were femoral neck, trochanter and total hip BMD, and the secondary outcomes were bone density measures at the radius and calcaneum. Outcomes were assessed at baseline, 12 months and 30 months using blinded operators. The analyses were performed on an intention-to-treat analysis. Mean femoral neck and trochanter BMD decreased in the control group [ 1.1%, 95% confidence interval (CI) -0.1% to -2.1% and -1.6%, 95% CI -0.4% to 2.7%], while no change occurred in the exercise group. Mean trochanter BMC decreased more in the control group (-7.7%, 95% CI -9.7% to -5.6% vs. -2.9%, 95% CI -5.3 to -0.9). There were six falls that resulted in fractures in the exercise group and 16 in the control group during the 30-month intervention (P=0.019). A significant bone loss occurred in both groups at the radius and calcaneum. In multivariate analysis, weight gain was associated with increased BMD and BMC at all femur sites both in the exercise group and in the pooled groups. In conclusion, impact exercise had no effect on BMD, while there was a positive effect on BMC at the trochanter. Exercise may prevent fall-related fractures in elderly women with low bone mass. PMID- 15889313 TI - Economic comparison of diagnostic approaches for evaluating osteoporosis in older women. AB - Of the technologies available, dual-energy X-ray absorptiometry of the hip or femoral neck (DXA-FN) is the best predictor of hip fractures. Diagnostic approaches utilizing measures of peripheral sites with office-based technology, such as calcaneal quantitative ultrasound (QUS), may reduce costs although clinical and economic outcomes have not been evaluated. The objective was to compare three approaches for diagnosing osteoporosis in older women. The design was a decision-analytic model using diagnostic measures and clinical and economic outcomes from the Study of Osteoporotic Fractures, a prospective cohort of older white women that measured BMD and QUS and assessed fracture outcomes. The setting and patients were a hypothetical cohort of older white women presenting for diagnosis of osteoporosis. For the diagnostic and treatment alternatives, three diagnostic approaches-DXA-FN alone, QUS alone and a sequential approach (first QUS, then DXA-FN for those with low values for QUS)-were compared to no diagnosis. The outcome measures were the number of women identified for treatment, number of hip fractures prevented following diagnosis and subsequent treatment, number of women needed to treat to prevent one hip fracture and total direct medical costs. The sequential approach identified fewer women to treat, prevented more hip fractures and incurred lower total costs than using DXA alone. Diagnosis with QUS alone identified more women to treat and incurred higher total costs than DXA alone under most conditions. Compared to other approaches for diagnosing osteoporosis, sequential use of QUS followed by DXA resulted in fewer women treated and lower total costs. PMID- 15889314 TI - Contribution of lean tissue mass to the urban-rural difference in bone mineral density. AB - While the urban-rural difference in bone mineral density (BMD) has been shown in some, but not all, Western populations, such a difference and the reason for the difference is largely unknown, particularly in developing countries. This cross sectional, epidemiologic study was designed to examine the hypothesis that differences in measures of body composition such as lean mass (LM) and fat mass (FM) contribute to the urban-rural difference in BMD. Lean mass, fat mass, lumbar spine and femoral neck BMD were measured by DXA (GE Lunar Corp, Wis.) in 411 urban (Bangkok city) and 436 rural (Khon Kaen province) Thai subjects, aged 20-84 years. Rural men and women had significantly higher LM and lower FM than their urban counterparts. In multiple linear regression analysis, age, LM, menopausal status (in women) and residence were independent determinants of BMD. After adjusting for age, menopause and LM, rural subjects were found to have significantly higher femoral neck BMD, but not lumbar spine BMD, than urban subjects. Furthermore, to alleviate the potential effect of multicolinearity of LM and FM, each rural subject was matched with each urban subject for FM and age, which resulted in 46 pairs of men and 91 pairs of women. In this matched-pair analysis, the femoral necks in rural men and women were, respectively, 7.3+/-2.1% (mean+/-SE; P<0.01) and 6.3+/-2.8% (P<0.02) higher than in urban men and women. The urban-rural difference in LM accounted for approximately 23 and 5% of the urban-rural difference in femoral neck BMD in men and women, respectively. These data are thus consistent with the hypothesis that the urban-rural difference in BMD at a weight-bearing site is in part associated with the urban-rural difference in lean mass. PMID- 15889315 TI - Changes in bone mineral density, body composition and biochemical markers of bone turnover during weight gain in adolescents with severe anorexia nervosa: a 1-year prospective study. AB - Osteoporosis is a serious complication of anorexia nervosa and in affected adolescents may result in a permanent deficit in bone mass. The pathophysiology of this bone disease has not been clearly defined. In this prospective study of 26 young women with anorexia nervosa aged 13-20 years (mean 16.5) we have measured changes in bone mineral density, total body composition and biochemical indices of bone turnover over 1 year. Over this period there was a mean weight gain of 10 kg and significant height gain with baseline and final values for body mass index of 14.2+/-1.7 and 17.6+/-2.3 kg/m2 (P<0.001). However, no significant changes were seen in bone mineral density in the spine or proximal femur during the study; total body bone mineral content was significantly higher than baseline at 3 months and 12 months (P=0.001 and P<0.0001), but total body bone mineral density at 3 months was significantly lower than baseline (P=0.003). Serum osteocalcin and bone-specific alkaline phosphatase values increased significantly and remained higher than baseline at all time points whereas urinary NTX/creatinine excretion showed a non-significant increase over the first 6 months of the study, but at 12 months, the mean value was significantly lower than baseline. Mean serum 25-hydroxyvitamin D levels showed a significant decrease at 6 months (P<0.05), but returned towards baseline thereafter. There was a significant increase in serum parathyroid hormone levels at all time points compared to baseline, these occurring within the normal range. These results indicate that although weight gain in young anorexics is associated with linear growth, bone mineral density does not increase. Whether this deficit can be corrected subsequently requires longer-term prospective studies. PMID- 15889318 TI - Trigeminocardiac reflex during skull base surgery: mechanism and management. AB - BACKGROUND: We study the occurrence and management of the trigeminocardiac reflex (TCR) during neurosurgical procedures for lesions of the skull base. METHOD: Two hundred patients underwent neurosurgical procedures for various skull base lesions and were evaluated retrospectively for the occurrence of the TCR during surgery. This phenomenon was defined as the onset of bradycardia lower than 60 beats/minute and hypotension with a drop in mean arterial blood pressure of 20% or more due to intra-operative manipulation or traction on the trigeminal nerve. FINDINGS: Sixteen patients (8%) had a TCR intra-operatively (7 vestibular schwannomas, 5 sphenoid wing meningiomas, 3 petroclival meningiomas, 1 intracavernous epidermoid cyst). In all 16 patients with a TCR the postoperative courses presented no complications that could be directly related to this intra operative phenomenon. CONCLUSIONS: Due to the intracranial course of the trigeminal nerve several surgical procedures at the anterior, middle and posterior skull base may elicit the trigeminocardiac reflex. Continuous monitoring of hemodynamic parameters allows the surgeon to interrupt surgical manoeuvres immediately upon the occurrence of the TCR. This technique is sufficient for the heart rate and the arterial blood pressure to return to normal levels without the necessity of additional anticholinergic medication. PMID- 15889316 TI - Factors associated with the lumbar spine and proximal femur bone mineral density in older men. AB - Bone mass is a major determinant of fracture, but there have been few comprehensive studies of the correlates of bone mineral density (BMD) in older men. The objective of the current cross-sectional analysis was to determine the factors associated with BMD of the lumbar spine and proximal femur in a large population-based sample of older men enrolled in The Osteoporotic Fractures in Men Study, "Mr.OS." We enrolled 5,995 men 65 years of age or older, 89% Caucasian, in Mr.OS at six US clinical centers. Demographic, medical and family history and lifestyle information was obtained by interview and physical function and anthropometric data by examination. Spine and hip BMD was measured using dual energy X-ray absorptimetry. The multivariable linear regression models predicted 19 and 10% of the overall variance in BMD of the femoral neck and spine, respectively. African-American men had 6 to 11% higher BMD than Caucasian men independent of multiple factors. Hip BMD declined with advancing age, while spine BMD increased. Body weight (per 10 kg) and self report of diabetes were each associated with 2 to 4% higher BMD, while history of a non-trauma fracture and current use of selective serotonin reuptake inhibitors, but not other antidepressants, were associated with at least 4% lower BMD. Both maternal and paternal histories of fracture were associated with 1.4-1.7% lower BMD. Osteoarthritis, physical activity, grip strength, alcohol intake, and dietary calcium were positively related to BMD, while a history of chronic lung disease, prostate cancer, and kidney stones was associated with lower BMD. Smoking, caffeine intake, and thiazide diuretics were not related to BMD in older men. A number of lifestyle and behavioral characteristics and medical conditions were associated with BMD in older men. Identification of these correlates could improve methods to identify men at risk for fracture and improve our understanding of fracture etiology. PMID- 15889319 TI - Brain oxygen monitoring: in-vitro accuracy, long-term drift and response-time of Licox- and Neurotrend sensors. AB - BACKGROUND: Oxygen tension sensors have been used to monitor tissue oxygenation in human brain for several years. The working principals of the most frequently used sensors, the Licox (LX) and Neurotrend (NT), are different, and they have never been validated independently for correct measurement in vitro. Therefore, we tried to clarify if the two currently available sensors provide sufficient accuracy and stability. METHOD: 12 LX oxygen tension sensors and NT sensors were placed into a liquid-filled tonometer chamber. The solution was kept at 37 +/- 0.2 degrees C and equilibrated with five calibration gases containing different O(2)- and CO(2)-concentrations. After equilibration, readings were taken for each gas concentration (accuracy test). Afterwards, the sensors were left in 3% O(2) and 9% CO(2) and readings were taken after 24, 48, 72, 96 and 120 hours (drift test). Thereafter, a 90% response time test was performed transferring sensors from 1% to 5% oxygen concentration and back, using pre-equilibrated tonometers. FINDINGS: All Licox oxygen probes [12] were used for this study. Two of 14 Neurotrend sensors did not calibrate, revealing a failure rate of 14% for NT. Oxygen tension during the accuracy test was measured as follows: 1% O(2) (7.1 mmHg): LX 6.5 +/- 0.4, NT 5.3 +/- 2.3 mmHg, 2% O(2) (14.2 mmHg): LX 12.9 +/- 0.6, NT 12.1 +/- 2.2 mmHg, 3% O(2) (21.4 mmHg): LX 19.8 +/- 0.7, NT 19.4 +/- 2.4 mmHg, 5% O(2) (35.8 mmHg): LX 33.4 +/- 1.0 mmHg, NT 33.5 +/- 2.9 mmHg, 8% O(2) (57.0 mmHg): 53.8 +/- 1.5, NT 53.6 +/- 3.3 mmHg. After 120 hours in 3% O(2) (21 mmHg), LX measured 19.8 +/- 1.9 mmHg, NT 17.9 +/- 4.7 mmHg. 90% response time from 1% to 5%/5% to 1% oxygen concentration was 129 +/- 27/174 +/- 26 sec for LX, 55 +/- 19/98 +/- 39 sec for NT. CONCLUSIONS: Both systems are measuring oxygen tension sufficiently, but more accurately with LX probes. NT sensors read significantly lower pO(2) in 1% O(2) and show an increasing deviation with higher oxygen concentrations which was due to two of twelve probes. A slight drift towards lower oxygen tension readings for both sensors but more pronounced for the NT does not impair long-term use. NT measures pCO(2) and pH very accurately. PMID- 15889320 TI - [Percutaneous endovascular aneurysm repair without arteriotomy (Perclose system)]. AB - BACKGROUND AND OBJECTIVE: It is common clinical practice to perform an arteriotomy for the endovascular treatment of infrarenal and thoracic aortic aneurysms. Instead we used the percutaneous endovascular Perclose device to perform the aneurysm repair without arterial cut-down. PATIENTS AND METHODS: The Perclose device contains four needles with two suture loops for closing the femoral artery access site. The sutures were deployed after the arterial puncture, before introduction of sheaths (diameter 12 - 27 F = 4 - 9 mm). After the procedure the sutures were used to close the puncture site. We attempted to achieve hemostasis with the Perclose system in 104 femoral arteries in 60 patients (7 females, mean age 69 +/- 12 years). The mean vessel diameter was 10 +/- 2 mm. RESULTS: The percutaneous graft implantation was successfully achieved in 58 of 60 patients. The graft could not be forwarded into the aorta in two cases because of calcified iliac arteries. The Perclose suture technique was successfully used in 97 femoral arteries. In one case a false aneurysm developed and in another case a secondary hemorrhage occurred. Seven patients needed additional manual compression to achieve complete hemostasis. A surgical repair was not necessary. The time to hospital discharge ranged from 4.5 hours to 32 days (median: 18 hours). 20 patients stayed longer than 24 h in the hospital, only 2 of them for reasons related to the puncture site. CONCLUSION: Closing the access site with the Perclose system is technically feasible and effective, even with large sheaths up to 27 F = 9 mm. This technique reduces the invasiveness of the endovascular repair of aortic aneurysms and decreases the length of hospital stay, i. e. it allows treatment in outpatients. PMID- 15889321 TI - [Intensified insulin therapy of type 2 diabetes mellitus: pre- or postprandial injection of aspart insulin?]. AB - BACKGROUND AND OBJECTIVE: Intensified insulin therapy using rapid acting insulin analogues is advocated in younger type 2 diabetic patients at risk of developing diabetic complications. Most patients prefer postprandial insulin injections. So far, however, there were no data on glycemic control by postprandial aspart insulin in patients with type 2 diabetes. PATIENTS AND METHODS: To compare blood glucose responsiveness to preprandial vs. postprandial aspart injections, a randomised open intraindividual cross-over trial was carried out. Blood glucose was measured before and one hour after the three main meals and at bedtime. 18 insulin-naive patients with type 2 diabetes (age, 60 +/- 3 years (mean +/- SEM), known duration of the disease, 7 +/- 2 years) participated at this study. RESULTS: Both with preprandial and postprandial injections of aspart insulin, the averages of the 7-point blood glucose profiles (8.27 +/- 0.50 vs. 8.5 +/- 0.61 mmol/l) were similar. With postprandial aspart insulin, however, 84 % of the blood glucose levels measured one hour after breakfast exceeded > 10 mmol/l in comparison to 38 % with preprandial aspart insulin (p < 0.05). Patients injected similar amounts of basal and aspart insulin on both experimental days (insulin glargin, 11 +/- 3 U, aspart insulin, 23 +/- 2 vs. 25 +/- 2 U/day, p = 0.4843). CONCLUSION: Both preprandial and postprandial insulin aspart can be allowed to well-controlled type 2 diabetic patients. However, patients will benefit from the recommendation to inject insulin aspart immediately before meal if food with a high glycemic index such as the continental breakfast is to be consumed. PMID- 15889322 TI - [Lethal sepsis and multi-organ failure after accidental injection of urine in a drug addict]. AB - HISTORY AND ADMISSION FINDINGS: A 33-year-old drug addict injected accidentally into the femoral vein 5 ml urine (kept in the refrigerator for 1 week for unanticipated drug-screening) which she mis-took for methadone. Soon after injection she was found confused with shivering. On admission she had a blood pressure of 90/60 mmHg, heart rate of 120/min and fever of 40 degrees C. INVESTIGATIONS: Laboratory tests showed consumption coagulopathy (DIC) as well as leukopenia. Blood cultures were positive for E.coli and Klebsiella pneumoniae. Infection with hepatitis C and HIV was excluded. Echocardiography revealed good ventricular function without signs of endocarditis. DIAGNOSIS AND TREATMENT: The patient developed a fulminant Gram-negative sepsis with DIC and multi-organ failure. She was intubated and mechanically ventilated. Acute renal failure required haemodialysis and ultrafiltration. Septic shock was treated with fluids and high doses of vasopressors. DIC was effectively treated with tranexamic acid, PPSB complex and fresh frozen plasma. The further course was complicated by septic emboli to the myocardium, liver, kidney and spleen. Laparotomy because of an acute abdomen revealed perforation of terminal ileum (mainly due to ischemic bowel lesions) requiring partial resection. The patient was weaned on day 18 with an initially uneventful further course. On day 25 the patient had a cardiac arrest with at first successful resuscitation but electromechanical dissociation proved fatal on day 26. CONCLUSION: Although serious systemic inflammation associated with i. v.-injection in drug addicts is rare, one has to be aware of severe complications with septic emboli. To our knowledge it is the first published case of i. v.-injection of urine followed by sepsis and multi-organ failure. Aggressive interdisciplinary treatment is required. PMID- 15889323 TI - [Leg ulcers: current aspects in diagnostic]. PMID- 15889324 TI - [Imaging techniques for showing the morphology and surface structure of extracranial internal carotid artery plaques]. AB - Methods of determining plaques in the extracranial internal carotid artery are ultrasound, computed tomography (CT) and magnetic resonance imaging (MRI). The goal of this overview is to introduce these methods and discuss their diagnostic and clinical significance. Ultrasound has the capability of showing the echogenity of the plaques. This method seems to demonstrate a relationship with the incidence of hemispheric symptoms. A histological association could only rarely be seen. The surface structure can only be clearly and correctly evaluated in individual cases under optimal circumstances. Computed tomography is capable of showing the calcium content and specific site of the plaques. However, lipid content and fibrous tissue cannot be deduced reliably. The currently available image resolutions limit a detailed imaging of surface characteristics. Magnetic resonance imaging provides, in vitro, a correct rendering of plaque composition and endoluminal surface structure. In vivo it is possible to recognize both the fibrous cap and necrotic core as well as intraplaque hemorrhage. Because of certain artefacts more detailed images are difficult to interpret. In conclusion, further clinical trials should reveal, whether these methods can be set in clinical routine to detect plaque surface and morphology for stratifying patients at risk of hemispheric symptoms during the spontaneous course as well as during carotid interventions. PMID- 15889325 TI - [Recycling of drugs in medical practice]. PMID- 15889326 TI - [Falling asleep disorders after taking coffee?]. PMID- 15889327 TI - [Curtailment of life expectancy by obesity]. PMID- 15889328 TI - [Therapy of amelanotic melanoma]. PMID- 15889331 TI - [Is fibromyalgia a disease? Regarding the contribution in DMW 23/2004]. PMID- 15889333 TI - [Patients with coronary disease and normal blood pressure: amlodipine vs. enalapril -- regarding the contribution in DMW 4/2005]. PMID- 15889335 TI - [Innovation instead of imitation -- news in drug therapy -- regarding the contribution in DMW 13/2005]. PMID- 15889336 TI - GMO, the last resort? PMID- 15889337 TI - [Prebiotic phosphate: a problem insoluble in water ? ]. AB - It is well-known that in water phosphate readily reacts with calcium, precipitating as insoluble apatite. How phosphorus could have been available for prebiotic reactions is still an open problem. We suggest that phosphorus containing compounds might have accumulated in a hydrophobic medium, since the absence of calcium ions would have prevented them from precipitating as apatite. Hydrophobic compounds may have been synthesized on the early Earth through the polymerization of methane or through Fischer-Tropsch-type reactions. Moreover, hydrophobic compounds would have been delivered to the early Earth by extraterrestrial infall. In previous articles (Morchio and Traverso [1999], Morchio et al. [2001]) we suggested that such hydrophobic material would have formed a hydrophobic layer on the surface of the sea, which would have provided an environment thermodynamically more suitable than water for the concentration and polymerization of organic molecules fundamental to life, particularly amino acids and (pyrimidine) bases. It may be hypothesized that elemental phosphorus or phosphorus-containing compounds (such as phosphite) deriving from volcanic eruptions would have ended up raining down into the hydrophobic layer, accumulating due to the absence of calcium ions, in an environment protected against hydrolysis. Phosphorus-containing compounds might have interacted with hydrophobic molecules in the layer giving rise to polymers. In particular, phosphite might have reacted with the hydrophobic amino acids, giving rise to phosphoamino acids, which, in turn, might have interacted with pyrimidine bases (relatively abundant in the layer) giving rise to peptides and oligonucleotide like polymers. Indeed, it has been experimentally shown (Zhou et al. [1996]) that, in an anhydrous organic medium (pyridine), dialkilphosphite reacts with amino acids to form phosphoamino acids, which interact with pyrimidine nucleosides to give nucleotides, short oligonucleotides and phosphoryl peptides. PMID- 15889338 TI - Origin of genetic instructions is presently unknown and unclear and requires scientific experimentation that is not readily possible. PMID- 15889339 TI - A new paradigma on the plant evolution: from a natural evolution to an artificial evolution? AB - After evidencing the great importance of plants for animals and humans in consequence of the photosynthesis, several considerations on plant evolution are made. One of the peculiar characteristics of the plant is the sessile property, due especially to the cell wall. This factor, principally, strengthened by the photosynthetic process, determined the particular developmental pattern of the plant, which is characterized by the continuous formation of new organs. The plant immobility, although negative for its survival, has been, in great part, overcome by the acquisition of the capacity of adaptation (plasticity) to the environmental stresses and changes, and the establishment of more adapted genotypes. This capacity to react to the external signals induced Trewavas to speak of "plant intelligence". The plant movement incapacity and the evolution of the sexual reproduction system were strongly correlated. In this context, the evolution of the flower in the Angiosperms has been particularly important to allow the male gamete to fertilize the immobile female gamete. Moreover, the formation of fruit and seed greatly improved the dispersal and conservation of the progeny in the environment. With the flower, mechanisms to favour the outcrossing among different individuals appeared, which are essential to increase the genetic variability and, then, the plant evolution itself. Although the Angiosperms seem highly evolved, the plant evolution is not surely finished, because many reported morpho-physiological processes may be still considered susceptible of further improvement. In the last years the relationships among humans, plants and environment are becoming closer and closer. This is due to the use of the DNA recombinant techniques with the aim to modify artificially plant characters. Therefore, the risk of a plant evolution strongly directed towards practical or commercial objectives, or "an artificial evolution", may be hypothesized. PMID- 15889340 TI - Darwinian criminality theory: a tragic chapter in history. AB - Darwinists once believed that individual human beings occasionally reverted, both physically and mentally, to a prehuman stage of evolutionary development. This person was called an atavistic criminal type. As a result of this belief, the focus in Darwinian criminology was on identifying the criminal type who should be imprisoned permanently to protect society, even if the particular offense committed was minor. Conversely, if a "non-criminal type" committed even a serious offense, it was an aberration, and therefore, they concluded, imprisonment would serve no purpose. Darwinian criminologists believed punishment must fit the criminal and not the crime. Criminologists widely adopted this theory to explain crime and, as a result, it influenced both public opinion and official policy. The "criminal physical type" stereotype is still very much with us, even though the theory of evolutionary throwbacks (atavisms) as a causative factor in criminality was empirically disproved decades ago. PMID- 15889341 TI - Do centrioles generate a polar ejection force? AB - A microtubule-dependent polar ejection force that pushes chromosomes away from spindle poles during prometaphase is observed in animal cells but not in the cells of higher plants. Elongating microtubules and kinesin-like motor molecules have been proposed as possible causes, but neither accounts for all the data. In the hypothesis proposed here a polar ejection force is generated by centrioles, which are found in animals but not in higher plants. Centrioles consist of nine microtubule triplets arranged like the blades of a tiny turbine. Instead of viewing centrioles through the spectacles of molecular reductionism and neo Darwinism, this hypothesis assumes that they are holistically designed to be turbines. Orthogonally oriented centriolar turbines could generate oscillations in spindle microtubules that resemble the motion produced by a laboratory vortexer. The result would be a microtubule-mediated ejection force tending to move chromosomes away from the spindle axis and the poles. A rise in intracellular calcium at the onset of anaphase could regulate the polar ejection force by shutting down the centriolar turbines, but defective regulation could result in an excessive force that contributes to the chromosomal instability characteristic of most cancer cells. PMID- 15889342 TI - Growth function of self-complementary circular codes. AB - In several papers Arques and Michel studied the maximal circular codes consisting of words of length 3 (or trinucleotides) on the genetic alphabet {A, C, G, T}. We present here some additional information on these codes. In particular, we study the growth function of the self-complementary circular codes and we prove that among them exactly 528 are maximal. PMID- 15889343 TI - From the ethology of unicellular eukaryotes to the locomotion of the living beings: meaning and evolution of the phenomenon. AB - A comparative reappraisal of the general problem of evolutionary trends and constraints of the locomotion phenomenon from prokaryotes to protozoa to metazoa was carried on. They elaborated different propulsive systems, different control systems of motion and different analysis systems of the stimuli. A general understanding of the locomotion phenomenon was reached and ciliate behaviour was positioned within the wider context of the evolution of biological displacement. PMID- 15889344 TI - The selfish karyotype. An analysis of the biological basis of morals. AB - An analysis is made of the kin-selection/group-selection debate on the issue of the biological basis of morals. The kin-selection view sees altruism, and morals in the case of humans, as resting solely on genetic factors; in fact, on this view, evolution itself is to take place only through genetic change -- a position which cannot be reconciled with our knowledge that species evolution involves karyotypic change. Morality is thought to stem from a particular gene which at one time was completely absent from the human population but which later entered and spread through it. On the generally accepted conception of kin selection, this 'altruistic' gene is to be responsible not only for apparently altruistic behaviour towards near relatives, but also for parental care. Thus this view, among other things, has the absurd consequence that there was a period directly after humans first came into existence during which we did not care for our young. In contrast, the group-selection view, as developed here, sees morality as stemming from the species' karyotype, and to be passed on karyotypically from species to species through evolution. As suggested by Darwin, morality is thus derived from our social instincts, and is fundamentally directed to members of one's tribe or community, not to members of one's family. In keeping with this perspective, it is suggested here that the biological basis of morals does not concern the continuation of each individual's gene line, but the continuing existence of the individual's community, and thus, indirectly, the survival of the human species, through supporting the continuing instantiation of its karyotype. PMID- 15889345 TI - A Prescribed Evolutionary Hypothesis. AB - I propose that phylogeny took place in a manner similar to that of ontogeny by the derepression of preformed genomic information which was expressed through release from latency (derepression) by the restructuring of existing chromosomal information (position effects). Both indirect and direct evidence is presented in support of the Prescribed Evolutionary Hypothesis. PMID- 15889346 TI - Antagonistic activity of a natural fungal population towards pathogenic bacteria. An in vitro study. AB - In the present work, we performed in vitro testing of 33 species of fungi of the subdivision Deuteromycotina isolated from water and sediment of the Kolubara River for antagonistic action towards 11 species of pathogenic bacteria. Of gram negative bacteria, the species most sensitive to metabolic fluid of the fungi were Proteus mirabilis, Salmonella enteritidis, and Shigella sonnei, while the most resistant were Klebsiella pneumoniae and Salmonella typhimurium. Of gram positive bacteria, the most sensitive species was Staphylococcus aureus, while the most resistant was Enterococcus faecalis. Of the tested fungi, Penicillium canescens, P. simplicissimum, P. thomii, Aspergillus fumigatus, A. ochraceus, and Fusarium culmorum exerted inhibitory action on the greatest number of species of pathogenic bacteria, while Verticillium lateritium, V. tenerum, Phoma humicola, and Botrytis cinerea had an inhibiting effect on the least number of species. PMID- 15889350 TI - Allan-Herndon-Dudley syndrome and the monocarboxylate transporter 8 (MCT8) gene. AB - Allan-Herndon-Dudley syndrome was among the first of the X-linked mental retardation syndromes to be described (in 1944) and among the first to be regionally mapped on the X chromosome (in 1990). Six large families with the syndrome have been identified, and linkage studies have placed the gene locus in Xq13.2. Mutations in the monocarboxylate transporter 8 gene (MCT8) have been found in each of the six families. One essential function of the protein encoded by this gene appears to be the transport of triiodothyronine into neurons. Abnormal transporter function is reflected in elevated free triiodothyronine and lowered free thyroxine levels in the blood. Infancy and childhood in the Allan Herndon-Dudley syndrome are marked by hypotonia, weakness, reduced muscle mass, and delay of developmental milestones. Facial manifestations are not distinctive, but the face tends to be elongated with bifrontal narrowing, and the ears are often simply formed or cupped. Some patients have myopathic facies. Generalized weakness is manifested by excessive drooling, forward positioning of the head and neck, failure to ambulate independently, or ataxia in those who do ambulate. Speech is dysarthric or absent altogether. Hypotonia gives way in adult life to spasticity. The hands exhibit dystonic and athetoid posturing and fisting. Cognitive development is severely impaired. No major malformations occur, intrauterine growth is not impaired, and head circumference and genital development are usually normal. Behavior tends to be passive, with little evidence of aggressive or disruptive behavior. Although clinical signs of thyroid dysfunction are usually absent in affected males, the disturbances in blood levels of thyroid hormones suggest the possibility of systematic detection through screening of high-risk populations. PMID- 15889351 TI - Infective dermatitis associated with the human T cell lymphotropic virus type I in Salvador, Bahia, Brazil. AB - BACKGROUND: Infective dermatitis associated with human T cell lymphotropic virus type I (HTLV-I) infection is a chronic, relapsing eczema of childhood. METHODS: Children, their mothers, and their siblings underwent serological testing for HTLV-I. Epidemiological data were collected from all seropositive children and their family members, and clinical and dermatological examinations were performed. Laboratory studies, including skin culture, and histopathological analyses were also performed. The diagnosis of infective dermatitis associated with HTLV-I (IDH) was made according to previously established criteria. RESULTS: All of the patients with cases that demonstrated clinical aspects of IDH were positive for HTLV-I. The median age of the children at the time of the first visit was 8.0 years (range, 2-14 years). The median duration of breastfeeding for 19 children was 22.5 months (range, 1-48 months). The lesions were erythematous, scaly, exudative, and crusted in all cases. The scalp, retroauricular areas, neck, and groin were the regions that were commonly affected. Cultures were positive for Staphylococcus aureus for 95% of the patients. The children were followed-up for a median of 3.0 years (range, 0.1-7 years), and 5 children developed HTLV-I-associated myelopathy/tropical spastic paraparesis. All of the children except 1 were treated with sulfamethoxazole-trimethoprim, and their lesions either improved greatly or completely disappeared. CONCLUSIONS: The present study demonstrates the severity of IDH in Bahia and confirms that its diagnosis is based almost exclusively on clinical aspects of the disease. Serological testing for HTLV-I and careful follow-up is recommended for all children with chronic, relapsing, severe eczema in regions where HTLV-I is endemic. PMID- 15889352 TI - Acute Plasmodium falciparum malaria following splenectomy for suspected lymphoma in 2 patients. AB - Two black African immigrants, with no history of recent travel outside France, received a diagnosis of a malignant lymphoproliferative disorder and splenomegaly, and they subsequently underwent splenectomy. A few weeks after surgery, both patients experienced an acute episode of Plasmodium falciparum malaria, so the initial diagnosis was corrected retrospectively and changed to hyperreactive malarial splenomegaly. These cases illustrate the difficulty in distinguishing hyperreactive malarial splenomegaly from malignant lymphoproliferative disorders and therefore underline the role of the spleen in the immune system's defense against malaria. PMID- 15889353 TI - Guidelines for the management of chronic kidney disease in HIV-infected patients: recommendations of the HIV Medicine Association of the Infectious Diseases Society of America. PMID- 15889354 TI - Relatively poor outcome after treatment of Clostridium difficile colitis with metronidazole. AB - BACKGROUND: Clostridium difficile is a frequent cause of serious nosocomial infection. Earlier reports have suggested that treatment with metronidazole cured nearly 90% of patients, with only a modest rate of recurrence of infection. In recent years, the rate of response to treatment with this drug has appeared to be much lower. METHODS: We undertook a prospective, observational study of 207 patients who were treated with metronidazole for C. difficile colitis. RESULTS: A total of 103 patients (50%) were cured by the initial course of therapy and had no recurrence of disease. Forty-six patients (22%) continued to have symptoms of colitis for > or = 10 days despite treatment, and 58 (28%) responded initially but had a recurrence within the ensuing 90 days. The mortality rate among patients who developed C. difficile colitis was 27%, and it was higher among patients who did not respond fully to an initial course of therapy, compared with those who did (33% vs. 21%; P < .05). CONCLUSIONS: Because of the relatively poor response to therapy, additional approaches to prevention and/or treatment of C. difficile colitis appear to be warranted. PMID- 15889355 TI - Increasing risk of relapse after treatment of Clostridium difficile colitis in Quebec, Canada. AB - BACKGROUND: Clinicians who treat patients with Clostridium difficile-associated diarrhea (CDAD) in Quebec, Canada, have noted an apparent increase in the proportion of patients who experience relapse. METHODS: To determine whether there was an increase in the frequency of treatment failure and of recurrence of CDAD after treatment, we reviewed data on cases that had been diagnosed in a hospital in the province of Quebec during the period 1991-2004. The frequency of recurrences within 60 days after the initial diagnosis was measured using Kaplan Meier analysis, and Cox regression was used for multivariate analysis. RESULTS: Among patients who had initially been treated with metronidazole, the proportion whose regimens were switched to vancomycin or for whom vancomycin was added because of a disappointing response did not vary between 1991 and 2002 (66 [9.6%] of 688 patients overall) but more than doubled in 2003-2004 (112 [25.7%] of 435; P < .001). Among 845 patients treated with metronidazole only, the 60-day probability of recurrence increased dramatically in 2003-2004 (47.2%), compared with 1991-2002 (20.8%; P < .001). During 1991-2002, the probabilities of recurrence were 20.0%, 13.8%, and 28.9% among individuals aged 0-17, 18-64, and > or = 65 years, respectively; during 2003-2004, the probabilities were 25.0%, 27.1%, and 58.4%, respectively. CONCLUSION: In 2003-2004, there was an increase in the proportion of patients with CDAD believed, by their attending physicians, to have experienced metronidazole treatment failure, as well as an increase in the frequency of post-metronidazole therapy recurrences, especially among elderly persons. PMID- 15889356 TI - Metronidazole for Clostridium difficile-associated disease: is it okay for Mom? PMID- 15889357 TI - Telavancin versus standard therapy for treatment of complicated skin and soft tissue infections due to gram-positive bacteria. AB - BACKGROUND: Telavancin, a novel lipoglycopeptide, exerts concentration-dependent, rapid bactericidal activity on account of its multiple mechanisms of action. Telavancin is highly active against gram-positive bacteria, including methicillin resistant Staphylococcus aureus (MRSA), vancomycin-intermediate, and vancomycin resistant strains. METHODS: We conducted a randomized, double-blind, controlled, phase-2 clinical trial. Patients > or = 18 years of age with a diagnosis of complicated skin and soft-tissue infection caused by suspected or confirmed gram positive organisms were randomized to receive either intravenously administered telavancin once daily or standard therapy (antistaphylococcal penicillin 4 times daily or vancomycin twice daily). RESULTS: For the study, 167 patients were randomized and received at least 1 dose of study medication. Success rates were similar in all analysis populations at the test-of-cure evaluation. Of patients with S. aureus infection at baseline (n = 102), 80% of the telavancin group were cured and 77% of the standard therapy group were cured. For patients with MRSA infection at baseline (n = 48), cure rates were 82% for the telavancin group and 69% for the standard therapy group. Microbiologic eradication in patients with MRSA infection was 84% for the telavancin group versus 74% for the standard therapy group. MIC90 values were lower for telavancin in all tested strains of S. aureus (< or = 0.25 ug/mL) compared with the MIC90 values for vancomycin and oxacillin. Similar proportions of patients discontinued therapy for adverse events in both treatment groups (approximately 5%). Fewer serious adverse events were reported in the telavancin group (4 events) than were for the standard therapy group (9). CONCLUSION: Clinical and microbiological results of this study support the further development of telavancin, especially for treatment of infection due to MRSA. PMID- 15889358 TI - Management of nonsevere pneumonia in military trainees with the urinary antigen test for Streptococcus pneumoniae: an innovative approach to targeted therapy. AB - BACKGROUND: The drug of choice for treatment of Streptococcus pneumoniae infection is generally a penicillin (including amoxicillin). Targeted therapy is, however, rarely used, because results of definitive diagnostic tests for pneumonia are not available for several days. Thus, broad-spectrum antibiotics are used for empirical treatment of pneumonia to cover both typical and atypical pathogens. Our purpose was to assess the usefulness of a strategy of targeted antimicrobial therapy based on the results of a rapid urinary antigen test for S. pneumoniae. METHODS: Military trainees with pneumonia were prospectively assigned to 2 groups: patients with positive urinary antigen test results who were treated with amoxicillin (1000 mg 3 times per day), and patients with negative urinary antigen test results who were treated with clarithromycin (500 mg 2 times per day). The duration of therapy was 5-10 days for both groups. RESULTS: A total of 219 evaluable patients were enrolled in the study. The most common causes of pneumonia were S. pneumoniae, Chlamydia pneumoniae, and Mycoplasma pneumoniae. Patients with positive urinary antigen test results had illness of greater severity at the time of study entry. Twenty-two percent of patients had positive urinary antigen test results (i.e., the amoxicillin group), and 78% had negative urinary antigen test results (i.e., the clarithromycin group). The clinical success rates were 94% for the clarithromycin group and 90% for the amoxicillin group (P = not significant). None of the patients who were classified as having treatment failure died. Resolution of clinical manifestations was slower for patients with pneumococcal pneumonia defined by a positive urinary antigen test result. CONCLUSIONS: The urine antigen test allowed targeted use of a penicillin (amoxicillin) for young immunocompetent individuals with nonsevere, community acquired pneumonia. Clarithromycin was highly effective against both S. pneumoniae pneumonia and pneumonia due to atypical pathogens. PMID- 15889359 TI - Low colonization prevalence of Staphylococcus aureus with reduced vancomycin susceptibility among patients undergoing hemodialysis in the San Francisco Bay area. AB - BACKGROUND: Staphylococcus aureus exhibits varying degrees of reduced vancomycin susceptibility, and strains with intermediate levels of resistance are thought to emerge by antibiotic selection of subpopulations in heterogeneously resistant precursor strains exposed to this antibiotic. We sought to determine the prevalence of and risk factors for carriage of potential heterogeneous vancomycin intermediate S. aureus (hVISA). METHODS: We prospectively observed a cohort of 211 patients undergoing hemodialysis and performed quarterly surveillance cultures for up to 2 years. We screened for reduced vancomycin susceptibility using brain-heart infusion agar with 4 microg/mL vancomycin. RESULTS: We identified 10 colonizing potential hVISA isolates recovered from 7 patients among both methicillin-susceptible and methicillin-resistant S. aureus strains. No confirmed hVISA isolates were identified; we can be 95% certain that the prevalence of confirmed hVISA carriage does not exceed 1.4%. When compared with noncolonized hemodialysis patients, neither vancomycin exposure, duration of hospitalization, nor any baseline clinical or demographic factor was found to predict colonization with potential hVISA on univariate analyses; increased number of months receiving hemodialysis was associated with potential hVISA colonization on multivariate analysis. CONCLUSIONS: Despite numerous published reports of S. aureus with reduced vancomycin susceptibility, carriage of these isolates remains a rare phenomenon. Given the unclear clinical significance of potential hVISA, it is not clear whether clinical laboratories should identify such strains, or how they should do so. PMID- 15889360 TI - Saccharomyces cerevisiae fungemia: an emerging infectious disease. AB - BACKGROUND: Saccharomyces cerevisiae is well known in the baking and brewing industry and is also used as a probiotic in humans. However, it is a very uncommon cause of infection in humans. METHODS: During the period of 15-30 April 2003, we found 3 patients with S. cerevisiae fungemia in an intensive care unit (ICU). An epidemiological study was performed, and the medical records for all patients who were in the unit during the second half of April were assessed. RESULTS: The only identified risk factor for S. cerevisiae infection was treatment with a probiotic containing Saccharomyces boulardii (Ultralevura; Bristol-Myers Squibb). This probiotic is used in Europe for the treatment and prevention of Clostridium difficile-associated diarrhea. The 3 patients received the product via nasograstric tube for a mean duration of 8.5 days before the culture result was positive, whereas only 2 of 41 control subjects had received it. Surveillance cultures for the control patients admitted at the same time did not reveal any carriers of the yeast. Strains from the probiotic capsules and the clinical isolates were identified as S. cerevisiae, with identical DNA fingerprinting. Discontinuation of use of the product in the unit stopped the outbreak of infection. A review of the literature identified another 57 cases of S. cerevisiae fungemia. Overall, 60% of these patients were in the ICU, and 71% were receiving enteral or parenteral nutrition. Use of probiotics was detected in 26 patients, and 17 patients died. CONCLUSIONS: Use of S. cerevisiae probiotics should be carefully reassessed, particularly in immunosuppressed or critically ill patients. PMID- 15889361 TI - Saccharomyces cerevisiae fungemia: an adverse effect of Saccharomyces boulardii probiotic administration. PMID- 15889362 TI - Electronic laboratory reporting for the infectious diseases physician and clinical microbiologist. AB - BACKGROUND: One important benefit of electronic health information is the improved interface between infectious diseases practice and public health. Electronic communicable disease reporting (CDR), given its legal mandate and clear public health importance, is a significant early step in the sifting and pooling of health data for purposes beyond patient care and billing. Over the next 5-10 years, almost all CDR will move to the internet. METHODS: This paper reviews the components of electronic laboratory reporting (ELR), including sifting through data in a laboratory information management system for reportable results, controlled "vocabularies" (e.g., LOINC, Logical Observation Identifiers Names and Codes [Regenstrief Institute], and SNOMED, Systematized Nomenclature of Medicine [College of American Pathologists]), the "syntax" of an electronic message (e.g., health level 7 [HL7]), the implications of the Health Insurance Portability and Accountability Act for ELR, and the obstacles to and potential benefits of ELR. RESULTS: There are several ways that infectious diseases physicians, infection control professionals, and microbiology laboratorians will participate in electronic CDR, including web-based case reporting and ELR, the direct, automated messaging of communicable disease reports from clinical lab information management systems to the appropriate public health jurisdiction's information system. CONCLUSIONS: ELR has the potential to make a large impact on the timeliness and the completeness of communicable disease reporting, but it does not replace the clinician's responsibility to submit a case report with important demographic and epidemiologic information. PMID- 15889363 TI - Destroying the life and career of a valued physician-scientist who tried to protect us from plague: was it really necessary? AB - Thomas Campbell Butler, at 63 years of age, is completing the first year of a 2 year sentence in federal prison, following an investigation and trial that was initiated after he voluntarily reported that he believed vials containing Yersinia pestis were missing from his laboratory at Texas Tech University. We take this opportunity to remind the infectious diseases community of the plight of our esteemed colleague, whose career and family have, as a result of his efforts to protect us from infection by this organism, paid a price from which they will never recover. PMID- 15889364 TI - Antiseptic "resistance": real or perceived threat? AB - Biocides (antiseptics, disinfectants, preservatives, and sterilants) are critical components of intervention strategies used in clinical medicine for preventing the dissemination of nosocomial diseases. Biocides are also used in community environments for personal hygiene and to prevent cross-contamination with foodborne pathogens. In vitro studies suggest that exposure to biocides results in reduced susceptibility to antibiotics and biocides by intrinsic or acquired mechanisms of resistance. In addition, microorganisms have adapted to biocide exposure by acquiring plasmids and transposons that confer biocide resistance, the same survival strategies to disseminate acquired mechanisms of resistance to biocides as they have for resistance to antibiotics. The scientific community must weigh the risks and benefits of using biocides in clinical and community environments, to determine whether additional precautions are needed to guide biocide development and use. At present, insufficient scientific evidence exists to weigh these risks, and additional research is needed to allow appropriate characterization of risks in clinical and community environments. PMID- 15889365 TI - Telithromycin: a ketolide antibiotic for treatment of respiratory tract infections. AB - Telithromycin, a recently approved ketolide antibiotic derived from 14-membered macrolides, is active against erythromycin-resistant pneumococci. Telithromycin has enhanced activity in vitro because it binds not only to domain V of ribosomal RNA (like macrolides do) but also to domain II. However, it is not active against streptococci and staphylococci with constitutive macrolide, lincosamide, and streptogramin B resistance. Telithromycin, available in an oral formulation, is approved by the US Food and Drug Administration for use in adults for treatment of (1) community-acquired pneumonia due to Streptococcus pneumoniae (including multidrug-resistant isolates), Haemophilus influenzae, Moraxella catarrhalis, Chlamydia pneumoniae, or Mycoplasma pneumoniae; (2) acute exacerbation of chronic bronchitis due to S. pneumoniae, H. influenzae, or M. catarrhalis; or (3) acute bacterial sinusitis due to S. pneumoniae, H. influenzae, M. catarrhalis, or methicillin- and erythromycin-susceptible Streptococcus aureus. It is not approved for treatment of tonsillitis, pharyngitis, or severe pneumococcal pneumonia. Unique visual adverse effects occurred in 0.27%-2.1% of patients receiving telithromycin therapy. Its enhanced activity against some common respiratory pathogens makes it a valuable addition to the available macrolides. PMID- 15889366 TI - Infected physicians and invasive procedures: safe practice management. AB - There is currently no public policy that provides guidance concerning whether and when physicians infected with hepatitis B virus (HBV), hepatitis C virus (HCV), and/or human immunodeficiency virus (HIV) can safely perform invasive procedures. A committee of experts in the fields of medicine, law, and biomedical ethics and 1 community member, aided by an advisory board, was established to produce recommendations for policy reform. An extensive literature review was conducted for these 3 infectious diseases, medicine, surgery, epidemiology, law, and bioethics to gather all relevant data. Special recommendations are made regarding the management of physicians who are infected with HIV, HBV, and/or HCV. This policy proposal includes a list of exposure-prone procedures and a decision chart that indicates under what conditions infected physicians can practice beyond the need for disclosure of their serological status. PMID- 15889367 TI - Seroprevalence of human T cell leukemia virus in HIV antibody-negative populations in rural Cameroon. AB - Seven hundred forty-seven serum samples collected from humans in 4 separate rural village areas in Cameroon were examined for antibody to human T cell leukemia viruses (HTLVs) by use of an enzyme immunoassay followed by a Western blot assay. Of the 88 serum samples that the enzyme immunoassay found to be repeatedly reactive, the HTLV status of 49 samples was confirmed by Western blot assay to be HTLV type I, and the status of 6 samples was confirmed to be HTLV type II. PMID- 15889368 TI - FcgammaRIIIb and complement component C7 codeficiency in a patient with recurrence of fulminant meningococcal septic shock. AB - Individuals with deficiencies of the late components of complement exhibit a susceptibility to the recurrence of meningococcal disease with a usually mild clinical presentation. We report the recurrence of fulminant meningococcal disease in a complement component C7-deficient patient. We found a total deficiency of FcgammaRIIIb on neutrophils, which could partially explain the unusually severe clinical presentation. PMID- 15889369 TI - Posaconazole as salvage therapy in patients with chronic granulomatous disease and invasive filamentous fungal infection. AB - Chronic granulomatous disease (CGD) is characterized by life-threatening bacterial and fungal infections. Treatment with posaconazole led to a complete response in 7 of 8 patients with CGD with invasive mold infections (7 proven cases and 1 possible case) after failure or intolerance of treatment with standard antifungal agents. In this preliminary study, salvage treatment with posaconazole was safe and effective. PMID- 15889370 TI - Treatment of multidrug-resistant tuberculosis during pregnancy: long-term follow up of 6 children with intrauterine exposure to second-line agents. AB - Treatment of gestational multidrug-resistant tuberculosis (MDR-TB) is controversial. We describe follow-up of 6 children exposed to second-line antituberculous agents in utero. Each child (average age, 3.7 years) underwent comprehensive clinical evaluation. One child had MDR-TB diagnosed. There was no evidence of significant late-presentation toxicity among the children. The results suggest that aggressive management of gestational MDR-TB may benefit both mother and child. PMID- 15889371 TI - A national survey of severe influenza-associated complications among children and adults, 2003-2004. AB - This report summarizes findings of a national survey conducted among infectious diseases consultants to assess complications associated with influenza during the 2003-2004 influenza season. The survey identified severe complications, including secondary infection with Staphylococcus aureus and deaths among children and adults, as well as perceived shortages in rapid diagnostic tests and influenza vaccine. PMID- 15889372 TI - Influenza: getting our attention. PMID- 15889374 TI - Choosing a study population for the evaluation of antifungal prophylaxis. PMID- 15889375 TI - Risk of congenital cytomegalovirus infection. PMID- 15889377 TI - Size of tuberculin skin test reaction, bacille Calmette-Guerin vaccination and tuberculosis in children. PMID- 15889379 TI - Classification of community-acquired endocarditis due to coagulase-negative staphylococci. PMID- 15889380 TI - Decreasing in vitro susceptibility of clinical Staphylococcus aureus isolates to vancomycin at the New York Hospital: quantitative testing redux. PMID- 15889381 TI - Lack of efficacy of voriconazole therapy in 2 patients with pulmonary fungus balls. PMID- 15889382 TI - CD4+ T lymphocytes and cryoglobulins in patients coinfected with HIV and hepatitis C virus: forgetting the hidden CD4+ cell subsets. PMID- 15889384 TI - Sepsis and meningitis due to Capnocytophaga cynodegmi after splenectomy. PMID- 15889385 TI - Neurotoxicity due to antimalarial therapy associated with misdiagnosis of malaria. PMID- 15889386 TI - Disentangling fetal and maternal susceptibility for pre-eclampsia: a British multicenter candidate-gene study. AB - The Genetics of Pre-Eclampsia (GOPEC) collaboration aims to identify genetic factors in U.K. families affected by pre-eclampsia. A number of genetic studies have reported associations with pre-eclampsia, but attempts to replicate these findings have yielded inconsistent results. We describe the results of extensive genotyping of seven candidate genes previously reported as conferring susceptibility to pre-eclampsia. Six hundred fifty-seven women affected by pre eclampsia and their families were genotyped at 28 single-nucleotide polymorphisms in the genes encoding angiotensinogen, the angiotensin receptors, factor V Leiden variant, methylene tetrahydrofolate reductase, nitric oxide synthase, and TNFalpha. Genotypes were analyzed by the transmission/disequilibrium test. Genotype risk ratios (GRRs) associated with maternal genotypes had a range of 0.70-1.16; GRRs associated with fetal genotypes had a range of 0.72-1.11. No GRR achieved the prespecified criteria for statistical significance (posterior probability >.05). We conclude that none of the genetic variants tested in this large study of strictly defined pre-eclamptic pregnancies confers a high risk of disease. The results emphasize the importance of conducting rigorously designed studies of adequate size to provide precise genetic risks with narrow confidence intervals, if overreporting of false-positive results is to be avoided. PMID- 15889387 TI - Novel alginate-poly(L-histidine) microcapsules as drug carriers: in vitro protein release and short term stability. AB - Spherical, smooth-surfaced and mechanically stable alginate-poly(L-histidine) (PLHis) microcapsules with narrow particle size distributions were prepared by incubating calcium alginate beads in aqueous solutions of PLHis. The in vitro release characteristics, drug loading and encapsulation efficiency of the microcapsules were investigated using bovine erythrocytes hemoglobin (Hb) as a model drug. The results showed that the concentration of Ca(2+) ions had a considerable effect on the drug loading, encapsulation efficiency and in vitro release behavior of the microcapsules. When the concentration of CaCl(2) in the PLHis solution was increased from 0 to 3.0% (w/v), the drug loading and encapsulation efficiency decreased significantly from 38.0 to 4.3% and from 92.9 to 8.0%, respectively, while the total cumulative release of Hb from microcapsules in phosphate buffered saline solution (PBS, pH 6.8) decreased from 96.2 to 72.8% in 24 h. No significant protein release was observed during 70 h of incubation in hydrochloric acid solution (pH 1.2). However, under neutral conditions (PBS, pH 6.8), the Hb was completely and stably released within 24-70 h. An explosion test showed that the stability of alginate-PLHis microcapsules depended strongly on the concentration of PLHis and the calcium ions in solution. [Diagram: see text] Microscopy photo of Hb-loaded alginate-PLHis microcapsules. PMID- 15889388 TI - pH effects on the complexation, miscibility and radiation-induced crosslinking in poly(acrylic acid)-poly(vinyl alcohol) blends. AB - The effect of pH on the complexation of poly(acrylic acid) with poly(vinyl alcohol) in aqueous solution, the miscibility of these polymers in the solid state and the possibility for crosslinking the blends using gamma radiation has been studied. It is demonstrated that the complexation ability of poly(vinyl alcohol) with respect to poly(acrylic acid) is relatively low in comparison with some other synthetic non-ionic polymers. The precipitation of interpolymer complexes was observed below the critical pH of complexation (pH(crit1)), which characterizes the transition between a compact hydrophobic polycomplex and an extended hydrophilic interpolymer associate. Films prepared by casting from aqueous solutions at different pH values exhibited a transition from miscibility to immiscibility at a certain critical pH, pH(crit2), above which hydrogen bonding is prevented. It is shown here that gamma radiation crosslinking of solid blends is efficient and only results in the formation of hydrogel films for blends prepared between pH(crit1) and pH(crit2). The yield of the gel fraction and the swelling properties of the films depended on the absorbed radiation dose and the polymer ratio. [Diagram: see text] SEM image of an equimolar PAA-PVA blend cast from a pH 4.6 solution. PMID- 15889389 TI - In vitro evaluation of biodegradable poly(butylene succinate) as a novel biomaterial. AB - Poly(butylene succinate) (PBSU) can be easily synthesized by condensation polymerization of the starting materials of succinic acid and butan-1,4-diol. It has good degradability and possesses excellent processability. Due to these advantages, PBSU was first evaluated in the present study for its potential application as a novel biomaterial. The in vitro biocompatibility of the PBSU was evaluated by monitoring proliferation and differentiation of osteoblasts cultured on the PBSU film substrates for different periods. The results showed that the PBSU was biocompatible as the osteoblasts could proliferate and differentiate on the PBSU plates. In addition, the hydrolytic degradation behavior of the PBSU films in the phosphate-buffered saline (PBS) was also investigated and the results suggested that the PBSU degraded in the PBS solution with the same behavior as that of the degradable poly(alpha-hydroxyesters). In addition to the biocompatibility and hydrolytic degradation, some physical properties, including hydrophilicity, and mechanical and thermal properties of the PBSU substrates, were also determined and the results revealed that the PBSU was hydrophilic and ductile with excellent processability. The biocompatibility of the PBSU, together with the advantages of hydrolytic degradability, hydrophilicity, and excellent processability, indicated that PBSU has the potential to be used as a biomaterial for tissue repair. [Diagram: see text] Alkaline phosphate activity of osteoblasts cultured on PBSU and TCPS substrates for different time periods. PMID- 15889390 TI - Biomolecular hybrid of a conducting polymer with DNA: morphology, structure, and doping behavior. AB - A poly(o-methoxyaniline) (POMA)/DNA [weight fraction of DNA (W(DNA)) = 0.45] hybrid was prepared by mixing their solutions in sterilized double distilled water. The solution turned green upon aging for a longer time, and the doping of POMA by DNA was complete after about 15 d of aging. The doping was confirmed from the UV-vis spectra where the 599 nm peak of POMA(EB) disappeared and a new peak for a pi to localized polaron band-transition appeared. With increasing aging time the new peak gradually shifted from 674 nm at 3 h to 820 nm at 15 d of mixing and thereafter it remained constant. The absence of a free carrier tail in the UV-vis spectra indicated a coiled structure of POMA in the complex. Circular dichroism spectra of the hybrid solution indicated that the DNA conformation (double helical structure) remained unchanged in the hybrid. The SEM micrograph of the freeze-dried hybrid showed a needle-like morphology of the DNA dispersed in a polymer matrix and it was completely different from the fibrillar network morphology of pure DNA in the solid state. The TEM micrograph indicated a homogeneous dispersion of DNA fibrils in the POMA matrix. The melting temperature of the POMA-DNA hybrid showed an increase compared to that of pure DNA by 5 degrees C, probably caused by an electrostatic interaction between the DNA anion and the POMA radical cation generated in the doping process. WAXS investigations revealed that the DNA crystal structure remained unchanged in the hybrid whereas the POMA crystal structure might be lost. An FT-IR study suggested that interaction occurred between the phosphoric acid group of DNA and a nitrogen atom of POMA through proton transfer from the OH group of the former. A schematic model of the POMA-DNA complex randomly anchoring POMA chains with the DNA molecule was proposed. The dc conductivity of the POMA-DNA complex was found to be ca. 10(-7) S . cm(-1). Hence, this work describes a procedure for making a DNA conducting polymer hybrid without changing the conformation and structure of DNA. [Diagram: see text] PMID- 15889391 TI - Microgel-based engineered nanostructures and their applicability with template directed layer-by-layer polyelectrolyte assembly in protein encapsulation. AB - A novel strategy for the fabrication of microcapsules is elaborated by employing biomacromolecules and a dissolvable template. Calcium carbonate (CaCO(3)) microparticles were used as sacrificial templates for the two-step deposition of polyelectrolyte coatings by surface controlled precipitation (SCP) followed by the layer-by-layer (LbL) adsorption technique to form capsule shells. When sodium alginate was used for inner shell assembly, template decomposition with an acid resulted in simultaneous formation of microgel-like structures due to calcium ion induced gelation. An extraction of the calcium after further LbL treatment resulted in microcapsules filled with the biopolymer. The hollow as well as the polymer-filled polyelectrolyte capsules were characterized using confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), and scanning force microscopy (SFM). The results demonstrated multiple functionalities of the CaCO(3) core - as supporting template, porous core for increased polymer accommodation/immobilization, and as a source of shell-hardening material. The LbL treatment of the core-inner shell assembly resulted in further surface stabilization of the capsule wall and supplementation of a nanostructured diffusion barrier for encapsulated material. The polymer forming the inner shell governs the chemistry of the capsule interior and could be engineered to obtain a matrix for protein/drug encapsulation or immobilization. The outer shell could be used to precisely tune the properties of the capsule wall and exterior. [Diagram: see text] Confocal laser scanning microscopy (CLSM) image of microcapsules (insert is after treating with rhodamine 6G to stain the capsule wall). PMID- 15889392 TI - A comparison of imputation methods in a longitudinal randomized clinical trial. AB - It is common for longitudinal clinical trials to face problems of item non response, unit non-response, and drop-out. In this paper, we compare two alternative methods of handling multivariate incomplete data across a baseline assessment and three follow-up time points in a multi-centre randomized controlled trial of a disease management programme for late-life depression. One approach combines hot-deck (HD) multiple imputation using a predictive mean matching method for item non-response and the approximate Bayesian bootstrap for unit non-response. A second method is based on a multivariate normal (MVN) model using PROC MI in SAS software V8.2. These two methods are contrasted with a last observation carried forward (LOCF) technique and available-case (AC) analysis in a simulation study where replicate analyses are performed on subsets of the originally complete cases. Missing-data patterns were simulated to be consistent with missing-data patterns found in the originally incomplete cases, and observed complete data means were taken to be the targets of estimation. Not surprisingly, the LOCF and AC methods had poor coverage properties for many of the variables evaluated. Multiple imputation under the MVN model performed well for most variables but produced less than nominal coverage for variables with highly skewed distributions. The HD method consistently produced close to nominal coverage, with interval widths that were roughly 7 per cent larger on average than those produced from the MVN model. PMID- 15889393 TI - Conformational changes of the amyloid beta-peptide (1-40) adsorbed on solid surfaces. AB - The conformational change of the 39-43 residues of the amyloid beta-peptide (Abeta) toward a beta-sheet enriched state promotes self-aggregation of the peptide molecules and constitutes the major peptide component of the amyloid plaques in Alzheimer patients. The crucial question behind the self-aggregation of Abeta is related to the different pathways the peptide may take after cleavage from the amyloid precursor proteins at cellular membranes. This work is aiming at determining the conformation of the Abeta (1-40) adsorbed on hydrophobic Teflon and hydrophilic silica particles, as model sorbent surfaces mimicking the apolar transmembrane environment and the polar, charged membrane surface, respectively. The mechanism by which the Abeta interacts with solid surfaces strongly depends on the hydrophobic/hydrophilic character of the particles. Hydrophobic and electrostatic interactions contribute differently in each case, causing a completely different conformational change of the adsorbed molecules on the two surfaces. When hydrophobic interactions between the peptide and the sorbent prevail, the adsorbed Abeta (1-40) mainly adopts an alpha-helix conformation due to H-bonding in the apolar part of the peptide that is oriented towards the surface. On the other hand, when the peptide adsorbs by electrostatic interactions beta-sheet formation is promoted due to intermolecular association between the apolar parts of the adsorbed peptide. Irrespective of the characteristics of the solid sorbent, crowding the surface results in intermolecular association between adsorbed molecules leading to a strong aggregation tendency of the Abeta (1-40). [Diagram: see text] CD spectra of Abeta (1-40) at pH 7: A) in solution ([Abeta]=0.2 mg.ml(-1)) freshly prepared (line) and after overnight incubation (symbols);B) on Teflon (Gamma=0.5 mg.m(-2)). PMID- 15889394 TI - Early atypical malignant transformation of a plexiform neurofibroma in a 4-year old boy with Neurofibromatosis 1. PMID- 15889396 TI - High rate treatment of terephthalic acid production wastewater in a two-stage anaerobic bioreactor. AB - The feasibility was studied of anaerobic treatment of wastewater generated during purified terephthalic acid (PTA) production in two-stage upflow anaerobic sludge blanket (UASB) reactor system. The artificial influent of the system contained the main organic substrates of PTA-wastewater: acetate, benzoate, and terephthalate. Three parallel operated reactors were used for the second stage, and seeded with a suspended terephthalate degrading culture, with and without additional methanogenic granular sludge (two different types). The first stage UASB-reactor was seeded with methanogenic granular sludge. Reactors were operated at 37 degrees C and pH 7. During the first 300 days of operation a clear distinction between the biomass grown in both reactor stages was obtained. In the first stage, acetate and benzoate were degraded at a volumetric loading rate of 40 g-COD/L . day at a COD-removal efficiency of 95% within the first 25 days of operation. No degradation of terephthalate was obtained in the first stage during the first 300 days of operation despite operation of the reactor at a decreased volumetric loading rate with acetate and benzoate of 9 g-COD/L . day from day 150. Batch incubation of biomass from the reactor with terephthalate showed that the lag-phase prior to terephthalate degradation remained largely unchanged, indicating that no net growth of terephthalate degrading biomass occurred in the first stage reactor. From day 300, however, terephthalate degradation was observed in the first stage, and the biomass in this reactor could successfully be enriched with terephthalate degrading biomass, resulting in terephthalate removal capacities of 15 g-COD/L . day. Even though no single reason could be identified why (suddenly) terephthalate degradation was obtained after such a long period of operation, it is suggested that the solid retention time as well the prevailing reactor concentrations acetate and benzoate may have played an important role. From day 1 of operation, terephthalate was degraded in the second stage. In presence of methanogenic granular biomass, high terephthalate removal capacities were obtained in these reactors (15 g-COD/L . day) after approximately 125 days of operation. From the results obtained it is concluded that terephthalate degradation is the bottleneck during anaerobic treatment of PTA wastewater. Pre-removal of acetate and benzoate in staged bioreactor reduces the lag-phase prior to terephthalate degradation in latter stages, and enables high rate treatment of PTA-wastewater. PMID- 15889397 TI - Sugar composition and FT-IR analysis of exopolysaccharides produced by microbial isolates from paper mill slime deposits. AB - Thirty exopolysaccharides (EPS) produced by bacteria isolated from biofilms or slimelayers from different paper and board mills in Finland, France and Spain were subjected to size exclusion chromatography and sugar compositional analysis. High performance size exclusion chromatography (HPSEC) analysis revealed that some samples were composed of several molecular weight populations. These samples were fractionated by size exclusion chromatography and pooled accordingly. Principal components analysis (PCA) of the sugar compositions of the different pools indicated the presence of glucans and mannans caused by insufficient removal of the carbon or nitrogen source (yeast extract) from the bacteria growth medium leading to an overestimation of the glucose and mannose level in the sample, respectively. From the point of view of slime problems the EPS populations are the most important for multivariate analysis. Four groups of EPSs have been recognized by PCA analysis: a group of EPSs produced by Enterobacter and related genera similar to the regularly reported colanic acid; a group of Methylobacterium EPSs having high galactose and pyruvate levels and two groups that showed less dense clusters produced by Bacillus and related genera, showing high mannose and/or glucose levels and Klebsiella EPSs that showed galactose with rhamnose as major characteristic sugar moieties. Fourier transform infrared spectroscopy (FT-IR) of the same samples followed by discriminant partial least squares regression (DPLS) and linear discriminant analysis (LDA) showed that, when used with a well-defined training set, FT-IR could be used clustering instead of time-consuming sugar composition analysis. The Enterobacter and Methylobacetrium EPS groups could be recognized clearly. However the fact that this could hardly be done for the other two groups in the dataset indicates the importance of a larger and well-defined training or calibration set. The potential to use FT-IR, as a tool for pattern recognition and clustering with respect to EPS structures produced by micro organisms isolated from a paper mill environment is discussed. PMID- 15889398 TI - Evaluation of a model for seeded isothermal batch protein crystallization. AB - Bulk protein crystallization, unlike small molecule crystallization, has found very limited use in biopharmaceutical manufacture. Most work in this area targets obtaining single large crystals for molecular structure determination by crystallography. Design and optimization of bulk crystallization for protein recovery and purification is much less common, and requires a mathematical model for analysis of laboratory data suitable for scale-up purposes. Traditionally, the crystal size distribution and method of moments is used to characterize the crystallization process. A simpler method is presented in this paper that utilizes the desupersaturation curve. The method uses an approach that does not require expensive instrumentation or characterization of the seed crystal size distribution. The method is extended to allow determination of both the mass deposition rate constant and the growth rate order from a single desuperaturation curve. Experimental data for the bulk crystallization of ovalbumin are used to validate the method. The rate constants and rate order obtained using the new method compare well with literature values. Scale-up is illustrated by prediction of the impact of changes in seed mass on protein crystallization. This new method offers a straightforward and low-cost alternative to traditional methods for the analysis and scale-up of protein crystallization data. PMID- 15889399 TI - Kinetic model of laccase-catalyzed oxidation of aqueous phenol. AB - Laccase from Trametes versicolor (EC 1.10.3.2) catalyzes the oxidation of aqueous phenol by oxygen and has demonstrated good potential for applications in various industrial and environmental processes. A kinetic model of this system has been developed to facilitate a better understanding of the mechanisms and rate limiting steps of enzyme-catalyzed transformation and to eventually assist in the choice and design of suitable reactor systems. A kinetic model was derived based on the differential and mass balance equations that describe the interactions of various forms of the enzyme with the aromatic substrate and oxygen. This model also incorporated an expression accounting for enzyme inactivation over time due to the reaction environment. The model was validated by comparing model predictions with experimental observations of phenol transformation and oxygen consumption over time at a variety of enzyme concentrations. Excellent agreement was found between experimental data and predictions of the kinetic model. Sensitivity analyses demonstrated that the reaction between oxidized-laccase and phenol was the rate-limiting step. PMID- 15889400 TI - Development of a novel adenovirus purification process utilizing selective precipitation of cellular DNA. AB - The use of recombinant adenoviral vectors for vaccination and gene therapy requires the development of purification processes that are cost-effective, scalable, and capable of robust host cell DNA clearance. An adenovirus purification process was developed which incorporates selective precipitation of host cell DNA, enabling a reduction in the use of costly nucleases and chromatographic resins while substantially improving DNA and protein clearance capabilities. In this work, three cationic detergents were evaluated for their capacity to selectively precipitate DNA from adenovirus-containing cell lysate. Parameters including pH, sodium chloride concentration, nonionic surfactant concentration, and cell density were investigated during development of the precipitation step. In a novel application, the cationic detergent domiphen bromide was found to have superior selectivity for host cell DNA. In addition, domiphen bromide-induced precipitation of adenovirus was shown to be reversible, which reduces the importance of mixing. Precipitation of DNA in the cell lysate coupled with primary clarification resulted in 3 logs of DNA clearance and improved impurity clearance in the subsequent ultrafiltration step. As a result, nuclease treatment and/or anion exchange chromatography can be eliminated, or included exclusively to improve process robustness. PMID- 15889401 TI - Lag-sensitive repetition suppression effects in the anterior parahippocampal gyrus. AB - Single-unit recording studies of monkeys have shown that neurons in perirhinal and entorhinal cortex exhibit activity reductions following stimulus repetition, and some have suggested that these "repetition suppression" effects may represent neural signals that support recognition memory. Critically, repetition suppression effects are most pronounced at short intervals between stimulus repetitions. Here, we used event-related functional magnetic resonance imaging (fMRI) to identify repetition suppression effects in the human medial temporal lobe and determine whether these effects are sensitive to the length of the interval between repetitions. Twenty-one participants were scanned while performing a continuous recognition memory task in which the interval between item repetitions was parametrically varied from 2 to 32 intervening items. We found evidence of repetition suppression in the anterior parahippocampal gyrus, but only when the repetition interval was relatively short. Moreover, bilateral hippocampal regions showed lag-sensitive repetition effects. Our results demonstrate that activity in the human medial temporal cortex, like that of monkeys, exhibits repetition suppression effects that are sensitive to the length of the interval between repetitions. PMID- 15889402 TI - Measurement of the severity of natural smooth surface (interproximal) caries lesions with polarization sensitive optical coherence tomography. AB - BACKGROUND AND OBJECTIVES: Previous studies have demonstrated that polarization sensitive optical coherence tomography (PS-OCT) can be used to image early dental caries. The purpose of this study was to compare the measured reflectivity of natural caries lesions found on smooth surfaces (interproximal lesions) with the mineral loss measured using digital microradiography (DM) in order to determine if PS-OCT can be used as a non-destructive method to measure the severity of dental decay and resolve the internal structure of caries lesions. MATERIALS AND METHODS: A PS-OCT system operating at 1,310 nm was used to acquire polarization resolved images of natural white spot lesions and pigmented lesions on the smooth surfaces of extracted teeth. The integrated reflectivity from lesion areas was compared to polarized light microscopic images (PLM) and to the integrated mineral loss from the same lesion area measured using high resolution DM. RESULTS: The frequently complex internal structure of caries lesions could be resolved with PS-OCT. Such structural information is potentially useful for determining whether or not the lesion is active and progressing or whether it has become arrested and remineralized and does not require intervention. There was a strong correlation between the integrated mineral loss of the caries lesions measured using DM and the integrated reflectivity in the perpendicular polarization axis of the PS-OCT system. CONCLUSIONS: The integrated reflectivity in the perpendicular polarization channel can be used to represent the severity of demineralization in natural early caries lesions. Therefore, PS-OCT has considerable potential as a non-destructive clinical probe of early caries lesions to assess their severity, monitor their progression over-time and potentially assess lesion activity. PMID- 15889403 TI - Processing content or location: distinct brain activation in a memory task. AB - Objects are defined by their content ("what") and by their location ("where"). In the visual system, processing of these two types of information is segregated into distinct anatomical and functional pathways. Using H2 (15)O positron emission tomography to measure cerebral blood flow, we examined the differences in processing of "what" when compared with "where" information in human memory. We found that the detection of deviations from a previously learned image sequence activates distinct brain regions depending on whether the image's content or its location has changed. When deviations of an image's content had to be detected, the left medial temporal lobe (MTL) activation increased. In contrast, detection of deviations from the learned locations of the objects induced increased activation in the right MTL and in the right parietal cortex. These data demonstrate distinct contributions of the left and right MTL to the processing of "what" vs. "where" in memory. PMID- 15889404 TI - Microstructural prototyping of ceramics by kinetic engineering: applications of spark plasma sintering. AB - The significance of kinetics on the development of microstructures in connection with sintering of ceramics is well recognized. In practice, however, it still remains a challenge to prepare designed microstructures via engineering the sintering kinetics because of an insufficient understanding of the different operative mechanisms that are in many cases overlapping. In this article the kinetic issues involved in sintering are described and discussed with respect to their potential for prototyping microstructures that yield desired properties. By exploiting and mastering the differences present in the kinetics of grain sliding, densification, chemical reactions, and grain growth, respectively, we have established processing principles for producing bulk ceramics with microstructures consisting of nano-sized grains, aligned grains, and/or non equilibrium-phase constitutions, and for achieving radically improved superplasticity in brittle ceramics. Although the studies quoted in this article were mainly carried out by spark plasma sintering, more general implications of them are expected, including efficient particle sliding, deformation-induced dynamic ripening, superplastic deformation-induced dynamic ripening, and non equilibrium integration. PMID- 15889405 TI - Chronic LPS exposure produces changes in intrinsic membrane properties and a sustained IL-beta-dependent increase in GABAergic inhibition in hippocampal CA1 pyramidal neurons. AB - Chronic inflammation has been reported to be a significant factor in the induction and progression of a number of chronic neurological disorders including Alzheimer's disease and Down syndrome. It is believed that inflammation may promote synaptic dysfunction, an effect that is mediated in part by pro inflammatory cytokines such as interleukin-1beta (IL-1beta). However, the role of IL-1beta and other cytokines in synaptic transmission is still poorly understood. In this study, we have investigated how synaptic transmission and neuronal excitability in hippocampal pyramidal neurons are affected by chronic inflammation induced by exposing organotypic slices to the bacterial cell-wall product lipopolysaccharide (LPS). We report that CA1 pyramidal neurons recorded in whole cell from slices previously exposed to LPS for 7 days had resting membrane potential and action potential properties similar to those of the controls. However, they had significantly lower membrane resistance and a more elevated action potential threshold, and displayed a slower frequency of action potential discharge. Moreover, the amplitude of pharmacologically isolated postsynaptic gamma-aminobutyric acid (GABA)ergic potentials, but not excitatory glutamatergic postsynaptic potentials, was significantly larger following chronic LPS exposure. Interestingly, co-incubation of the IL-1 receptor antagonist (IL 1Ra) concurrently with LPS prevented the increase in GABAergic transmission, but not the reduction in intrinsic neuronal excitability. Finally, we confirmed that LPS dramatically increased IL-1beta, and IL-1beta-dependent IL-6 levels in the culture medium for 2 days before returning to baseline. We conclude that CA1 pyramidal neurons in slices chronically exposed to LPS show a persistent decrease in excitability due to a combined decrease in intrinsic membrane excitability and an enhancement in synaptic GABAergic input, the latter being dependent on IL 1beta. Therefore, chronic inflammation in hippocampus produces IL-1beta-dependent and -independent effects in neuronal and synaptic function that could contribute significantly to cognitive disturbances. PMID- 15889406 TI - Assessment of patient radiation doses during transcatheter closure of ventricular and atrial septal defects with Amplatzer devices. AB - The purpose of this study was to estimate the radiation dose to which children are exposed during cardiac catheterizations for the treatment of ventricular and atrial septal defects. Radiation doses were estimated for 46 children aged 1-18 years. These children were treated for secundum atrial septal defects (ASD group) for perimembranous ventricular septal defects (VSD group) or underwent a routine diagnostic catheterization (diagnostic group). Thermoluminescent dosimeters (TLDs) were attached in locations, representing the lateral entrance dose, the posterior entrance dose, the thyroid dose, and the gonad dose, respectively. A dose area product (DAP) meter was also attached externally on the posterior anterior (PA) tube to give a direct value in cGy cm(2) for each procedure. The patient's entrance dose from the PA field ranged from 1.5 to 185.0 mGy for all patients, while the lateral entrance dose varied from 0.9 to 204 mGy. Radiation exposure to the thyroid and the gonads was found to vary from 0.4 to 8.3 and 0.1 to 2.1 mGy, respectively. The DAP meter recorded DAP values for the posterior tube, between 46 and 3,700 cGy cm(2). The mean effective dose was found to be 7.7, 16.2, and 33.3 mSv for the diagnostic, the ASD, and the VSD group, respectively. Very strong correlation was found between the DAP values and the entrance radiation dose measured with TLDs. The mean entrance dose received from therapeutic cardiac catheterizations using the Amplatzer devices was found approximately twice the dose received from a diagnostic one. Even for the most complex procedures, the maximum entrance dose was at least 10 times lower than the threshold, associated with skin erythema. PMID- 15889407 TI - Effects of ionic strength on lysozyme uptake rates in cation exchangers. I: Uptake in SP Sepharose FF. AB - Fluorescence scanning confocal microscopy was used in parallel with batch uptake and breakthrough measurements of transport rates to study the effect of ionic strength on the uptake of lysozyme into SP Sepharose FF. In all cases the adsorption isotherms were near-rectangular. As described previously, the intraparticle profiles changed from slow-moving self-sharpening fronts at low salt concentration, to fast-moving diffuse profiles at high salt concentration, and batch uptake rates correspondingly increased with increasing salt concentration. Shrinking core and homogeneous diffusion frameworks were used successfully to obtain effective diffusivities for the low salt and high salt conditions, respectively. The prediction of column breakthrough was generally good using these frameworks, except for low-salt uptake results. In those cases, the compressibility of the stationary phase coupled with the shrinking core behavior appears to reduce the mass transfer rates at particle-particle contacts, leading to shallower breakthrough curves. In contrast, the fast uptake rates at high ionic strength appear to reduce the importance of mass transfer limitations at the particle contacts, but the confocal results do show a flow rate dependence on the uptake profiles, suggesting that external mass transfer becomes more limiting at high ionic strength. These results show that the complexity of behavior observable at the microscopic scale is directly manifested at the column scale and provides a phenomenological basis to interpret and predict column breakthrough. In addition, the results provide heuristics for the optimization of chromatographic conditions. PMID- 15889408 TI - Monolayer-derivative functionalization of non-oxidized silicon surfaces. AB - This article describes a variety of monolayers anchored directly onto silicon surfaces without an oxide interlayer, their formation mechanisms, their technological applications, and our personal views on the future prospects for this field. The chemical modification of non-oxidized silicon surfaces utilizing monolayers was first reported in 1993. The basic finding that a non-oxidized silicon surface could be neutralized with alkyl chains through direct covalent linkage, i.e., silicon-carbon, has offered chemical scientists ease of handling even in an ambient environment and, thus, research has been predictably focused on forming anti-stiction coating films for nano- and micro-electromechanical systems (NEMS/MEMS). Such surface reforming has also been achieved by using other monolayers, which form interfacial bonds, e.g., silicon-nitrogen and silicon oxygen. The resultant monolayer surfaces are useful for silicon-based applications including molecular electron transfer films, monolayer templates, molecular insulators, capsulators, and bioderivatives. Such monolayers are applicable not only for surface modification, but also for manipulating individual nanomaterials. By modifying the terminal groups of monolayers with nanomaterials including nanocrystals and biomolecules, the nanomaterials can remarkably be immobilized directly onto non-oxidized silicon surfaces based on the formation mechanisms of the monolayer. Such immobilizations will revolutionize the analysis of the specific features and capabilities of individual nanomaterials. Furthermore, the path will be opened for the development of more advanced monolayer-derived chip technology. To achieve this goal, it is extremely important to thoroughly understand the functionalization processes on silicon, since the resultant internal structures and properties of monolayer-derivative silicon may strongly depend on their course of formation. PMID- 15889409 TI - The surface chemistry of heterogeneous catalysis: mechanisms, selectivity, and active sites. AB - The role of chemical kinetics in defining the requirements for the active sites of heterogeneous catalysts is discussed. A personal view is presented, with specific examples from our laboratory to illustrate the role of the chemical composition, structure, and electronic properties of specific surface sites in determining reaction activity and selectivity. Manipulation of catalytic behavior via the addition of chemical modifiers and by tuning of the reaction conditions is also introduced. PMID- 15889410 TI - Unstable Robertsonian translocations der(13;15)(q10;q10): heritable chromosome fission without phenotypic effect in two kindreds. AB - Robertsonian translocations (RTs) are amongst the most common chromosome abnormalities, but being essentially balanced are not usually associated with phenotypic abnormality. Despite being dicentric, RTs are almost always transmitted stably through cell division without chromosome breakage. We have investigated spontaneous fission of der(13;15)(q10;q10) chromosomes in eight individuals from two unrelated kindreds with a view to assessing clinical significance and to seek an explanation for the peculiar heritable instability displayed by these chromosomes. In Family 1, fission products were observed in five members in three generations. The instability was observed in cells derived from chorionic villus and lymphocytes. In Family 2, the same phenomenon was observed in amniocytes from two separate pregnancies and maternal blood lymphocytes. Detailed FISH analysis of these RTs showed them to be dicentric with an unremarkable pericentromeric structure. Notably, combined immunofluoresence and FISH analysis showed the presence of the centromere-specific proteins CENP-A and CENP-E, consistent with functional dicentricity in >75% of cells analyzed. The fission products are, therefore, presumed to be the result of sporadic, bipolar kinetochore attachment, anaphase bridging with resultant inter centromeric breakage in a small proportion of mitoses. None of the eight carriers shows phenotypic abnormality and therefore, for prenatal counseling purposes, there appears to be no increased specific risk associated with this phenomenon. PMID- 15889411 TI - Living with a hereditary disease: persons with muscular dystrophy and their next of kin. AB - This qualitative study describes conceptions and experiences of the hereditary aspect of muscular dystrophy (MD) from both the patients' and the next of kin's perspective. Different diagnoses of MD are included: dystrophia myotonica, myopathia distalis tarda hereditaria, Becker MD, facioscapulohumeral MD, limb girdle MD, Emery-Dreifuss and undetermined proximal MD (Duchenne MD is not included). Interviews were conducted with 46 persons with MD and 36 next of kin. The interviews were subjected to inductive content analysis. Only two in each group did not spontaneously mention anything related to the fact that MD is disease with dominant or recessive inheritance. It was found that heredity has a prominent place in the thoughts and feelings of the family. These thoughts were classified as Becoming aware of MD and its hereditary nature, looking into the pedigree, acquiring an understanding of MD, thoughts about genetic testing, interpreting the risk, whether to have children or not, feelings related to the future, and feelings of responsibility and guilt. Families with MD need medical information and the opportunity for genetic testing as well as support and counseling in coming to terms with living with a hereditary disease, whether or not that includes a decision to take a test. PMID- 15889412 TI - Dual substrate recognition of aminotransferases. AB - Pyridoxal 5'-phosphate-dependent aminotransferases reversibly catalyzes the transamination reaction in which the alpha-amino group of amino acid 1 is transferred to the 2-oxo acid of amino acid 2 (usually 2-oxoglutarate) to produce the 2-oxo acid of amino acid 1 and amino acid 2 (glutamate). An aminotransferase must thus be able to recognize and bind two kinds of amino acids (amino acids 1 and 2), the side chains of which are different in shape and properties, from among many other small molecules. The dual substrate recognition mechanism has been discovered based on three-dimensional structures of aromatic amino acids, histidinol phosphate, glutamine:phenylpyruvate, acetylornithine, and branched chain amino acid aminotransferases. There are two representative strategies for dual substrate recognition. An aromatic amino acid aminotransferase prepares charged and neutral pockets for acidic and aromatic side chains, respectively, at the same place by a large-scale rearrangement of the hydrogen-bond network caused by the induced fit. In a branched-chain aminotransferase, the same hydrophobic cavity implanted with hydrophilic sites accommodates both hydrophobic and acidic side chains without side-chain rearrangements of the active-site residues, which is reminiscent of the lock and key mechanism. Dual substrate recognition in other aminotransferases is attained by combining the two representative methods. PMID- 15889413 TI - FXTAS, SCA10, and SCA17 in American patients with movement disorders. PMID- 15889414 TI - Green tea: Health benefits as cancer preventive for humans. AB - Green tea is an acknowledged cancer preventive in Japan. The aim of this review article is to develop the concept of cancer prevention with green tea beverage for humans, which has largely been our exclusive research territory. This paper briefly reviews several topics, beginning with the introduction of our initial work on penta-O-galloyl-beta-D-glucose and (-)-epigallocatechin gallate (EGCG), the main constituent of green tea extract. The mechanisms of EGCG action, particularly the reduction of TNF-alpha are discussed, and we show how use of 3H EGCG revealed a wide range of target organs for cancer prevention. The results of an epidemiological study in Saitama Prefecture allowed us to determine the cancer preventive amount of green tea-10 Japanese-size cups per day, about 2.5 g green tea extract-which made it possible for us to introduce the two-stage strategy of cancer prevention with green tea. The first stage is the delay of cancer onset for the general population. The second stage is the prevention of recurrence of cancer for patients following cancer treatment. Combination cancer prevention with green tea and cancer preventive drugs is proving especially beneficial for Japanese, who drink green tea every day. And finally, the stimulating comments of Prof. Jim Watson have encouraged green tea scientists. PMID- 15889415 TI - Array comparative genomic hybridization analysis of a familial duplication of chromosome 13q: a recognizable syndrome. AB - We report on a family with six persons in three generations who have mild mental retardation, behavioral problems, seizures, hearing loss, strabismus, dental anomalies, hypermobility, juvenile hallux valgus, and mild dysmorphic features. Classical cytogenetic analysis showed a partial duplication of chromosome 13q, array comparative genomic hybridization showed the duplication to span approximately 21 Mb, ranging from chromosome band 13q21.31 to 13q31.1. The relatively mild presentation of this large duplication may be explained by the relative paucity of genes in the chromosome region involved. Genotype-phenotype correlations in patients with similar partial 13q duplications are inconsistent. Emerging cytogenetic techniques will allow more reliable genotype-phenotype correlations. PMID- 15889416 TI - Floating-Harbor syndrome complicated by tethered cord: a new association and potential contribution from growth hormone therapy. AB - Floating-Harbor syndrome is a rare syndrome with short stature, severely delayed bone age, typical facies and delay in expressive speech. Structural malformations are uncommon, and tethered cord or other forms of spinal dysraphism have not previously been reported. We report on a case of Floating-Harbor syndrome, complicated by tethered cord and discuss the possibility that growth hormone therapy may contribute to the development of symptoms of this malformation. PMID- 15889417 TI - Relationship between polymorphisms in genes involved in homocysteine metabolism and maternal risk for Down syndrome in Brazil. AB - Associations between specific alleles of genes encoding enzymes in the methionine/homocysteine pathway and plasma homocysteine levels have been examined in different populations. In the present study, we determined polymorphisms of MTHFR A222V (677C > T), MTHFR E429A (1298A > C), MTRR I22M (66A > G), MTR D919G (2756A > G), and CBS 844ins68 and total plasma homocysteine levels (tHcy) among 154 mothers of children with Down syndrome (DS) and 158 control mothers from Brazil. Homocysteine levels were higher among DS mothers compared to control groups (10.437 vs. 8.600 respectively, P = 0.002). Only the 677T allele was associated with altered levels of tHcy in the case group (F((2,153)) = 5.300; P = 0.006), primarily when homozygous. In the control group, the association of the TT genotype with higher levels of tHcy showed borderline significance (F((2,157)) = 2.974; P = 0.054). All genotype distributions were similar in the two groups (P > 0.05), but the frequency of the 677T allele in the case group was significantly higher (chi(2) = 3.862; DF = 1; P = 0.049; OR = 1.437 (1.001-2.062)). Although the 677T allele is associated with increased homocysteine levels, its presence has only a modest impact as an independent risk factor for DS. All the other polymorphisms did not show an association with risk for the syndrome, when evaluated separately (P > 0.05). However, when the presence of 677T, 1298C, 2756G, 66G, and 844ins68 alleles were evaluated together, the mothers of children with DS tend to have a higher number of uncommon alleles than the mothers with no previous affected child. PMID- 15889418 TI - Higher frequency of uncommon 1.5-2 Mb deletions found in familial cases of 22q11.2 deletion syndrome. AB - Familial 22q11.2 deletions have been reported as a 6%-28% of the total affected cases of 22q11.2 microdeletion syndrome (del22q11.2). Different deletion genotypes have been described for this disorder, with a predominant 3 Mb deletion present in 90% of the cases, a less common 1.5-2 Mb deletion in 8%, and atypical smaller deletions in 2%. We have studied 15 cases of del22q11.2 from 6 families (two of them three-generation families) that were previously diagnosed through FISH. We have sized the deleted region by allele genotyping of 12-16 polymorphic markers in all cases, and we have found three families affected with the 1.5-2 Mb deletion, two affected with the 3 Mb deletion, and one in which the deletion size could not be determined. This predominance of the smaller 1.5-2 Mb deletions in our familial cases differs from the minor frequency observed in sporadic cases of del22q11.2. This finding suggests that small deletions are more linked to familial inheritance than large ones, possibly due to psychosocial or biological factors associated with differences in the phenotype. Deletion sizing on routine diagnosis may help characterizing the inheritability of 22q11.2 microdeletion syndrome. PMID- 15889419 TI - Another case of interstitial del(12) involving the proposed cardio-facio cutaneous candidate region. AB - Recent reports of patients with interstitial deletions involving the long arm of chromosome 12 have led to the proposal of a candidate region for the cardio-facio cutaneous syndrome (CFCS) at (12)(q21.2q22). We now report a patient with an interstitial deletion, del(12)(q21.1q21.3) that overlaps the proposed critical region. The patient is an 11-year-old female with developmental delay. Her growth was normal but she is microcephalic with low set ears. In common with other patients with deletions in this region, she had fine, sparse head and eyebrow hair and a hyperkeratotic follicular rash, which involved her face and limbs. She does not have the diagnostic features of the CFC syndrome. PMID- 15889420 TI - Long-term follow-up in a patient with metatropic dysplasia. PMID- 15889421 TI - Tricalcium phosphate and glutaraldehyde crosslinked gelatin incorporating bone morphogenetic protein--a viable scaffold for bone tissue engineering. AB - Bone defects caused by various etiologies must be filled with suitable substances to promote bone repair. Autogenous iliac crest graft is most frequently used, but is often associated with morbidities. Several bone graft substitutes have been developed to provide osteoconductive matrices as well as to enhance osteoinductivity. A tricalcium phosphate and glutaraldehyde crosslinked gelatin (GTG) scaffold, incorporated with bone morphogenetic proteins (BMPs), was developed to provide an alternative mean of bone tissue engineering. This study investigated differences between GTG and BMP-4 immobilized GTG (GTG-BMP) scaffolds on neonatal rat calvaria osteoblast activities. The GTG scaffold possessed an average pore size of 200 microm and a porosity of 75%. HE staining revealed uniform cell distribution throughout the scaffold 24 h post cell seeding. Alkaline phosphatase (ALP) activity of the GTG samples increased initially and then stabilized at 3 weeks postseeding. ALP activity of the GTG-BMP samples was similar to that of the GTG samples in the second and third weeks, but it continued increasing and became significantly greater than that of the GTG samples by the fourth week. Gla-type osteocalcin (Gla-OC) activity of the GTG-BMP samples was initially lower, but also became significantly greater than that of the GTG samples by the fourth week. An HE stain revealed greater numbers of attached cells and a richer matrix deposits in the GTG-BMP samples. A von Kossa stain showed larger mineralizing nodules, in greater numbers, after 4 weeks of in vitro cultivation. These findings suggest that the GTG scaffold provides an excellent porous structure, conductive to greater cell attachment and osteoblast differentiation, and that utility can be significantly enhanced by the inclusion of BMPs. A GTG-BMP scaffold holds promise as a superior bioactive material for bone tissue engineering. PMID- 15889422 TI - Role of thyroid hormone in the initiation of EGF (epidermal growth factor) expression in the sublingual gland of the postnatal mouse. AB - The effect of triiodo-L-thyronine (T3) and propylthiouracil (PTU) on the initiation of epidermal growth factor (EGF) expression in the sublingual glands (SLGs) of postnatal mice was investigated by indirect enzyme-labeled and immunogold antibody methods for light and electron microscopy, respectively. In normal males, EGF immunoreactivity first appeared in a few scattered granular cells of striated ducts (SDs) at 5 weeks of age, and the immunoreactive cells had increased in number at 6 weeks of age. No EGF expression was observed in the glands of females at any ages examined. When T3 (1 mg/kg body weight) was given to males every other day for 2 weeks before examination, EGF expression began earlier; the immunoreactive granular cells were first detected at 4 weeks of age, and at later ages they were markedly increased in number compared to those of normal males. Moreover, T3 was capable of inducing EGF in the female glands. After T3 was administered to females in the same manner as in males, a few immunoreactive cells were first detected at 5 weeks of age, and increased numbers were detected at later ages. By contrast, when PTU (1 mg/kg body weight) was given to male mice every other day for 2 weeks before examination, the EGF immunoreactive cells were markedly decreased in number compared to those of normal males of the same age. Electron microscopy revealed that many SD cells contained secretory granules, and that these cells constituted the granular striated tubule (GST) in a portion of SDs, but they were undetectable by light microscopy, because their secretory granules were minimal in size and few in number. Gold-labeling of EGF was confined to the secretory granules of scattered granular cells, whose secretory granules were far larger in size and more abundant than those of the GST cells. These results suggest that thyroid hormone is essential to differentiation of the cellular phenotype of GST precursor cells into typical granular cells (detectable by light microscopy) that express EGF in the mouse SLG, showing a close resemblance to the submandibular granular convoluted tubule cells. PMID- 15889423 TI - High-resolution simultaneous three-photon fluorescence and third-harmonic generation microscopy. AB - In recent years, nonlinear laser scanning microscopy has gained much attention due to its unique ability of deep optical sectioning. Based on our previous studies, a 1,200-1,300-nm femtosecond laser can provide superior penetration capability with minimized photodamage possibility. However, with the longer wavelength excitation, three-photon-fluorescence (3PF) would be necessary for efficient use of intrinsic and extrinsic visible fluorophores. The three-photon process can provide much better spatial resolution than two-photon-fluorescence due to the cubic power dependency. On the other hand, third-harmonic-generation (THG), another intrinsic three-photon process, is interface-sensitive and can be used as a general structural imaging modality to show the exact location of cellular membranes. The virtual-transition characteristic of THG prevents any excess energy from releasing in bio-tissues and, thus, THG acts as a truly noninvasive imaging tool. Here we demonstrated the first combined 3PF and THG microscopy, which can provide three-dimensional high-resolution images with both functional molecule specificity and sub-micrometer structural mapping capability. The simultaneously acquired 3PF and THG images based on a 1,230-nm Cr:forsterite femtosecond laser are shown with a Hoechst-labeled hepatic cell sample. Strong 3PF around 450 nm from DNA-bounded Hoechst-33258 can be observed inside each nucleus while THG reveals the location of plasma membranes and other membrane based organelles such as mitochondria. Considering that the maximum-allowable laser power in common nonlinear laser microscopy is less than 10 mW at 800 nm, it is remarkable that even with a 100-mW 1,230-nm incident power, there is no observable photo damage on the cells, demonstrating the noninvasiveness of this novel microscopy technique. PMID- 15889424 TI - Quantitative measurement of the resolution and sensitivity of confocal microscopes using line-scanning fluorescence correlation spectroscopy. AB - Spatial resolution and the sensitivity to detect a fluorophore are the two most important optical parameters that characterize a confocal microscope. However, these are rather difficult to estimate quantitatively. We show that fluorescence correlation spectroscopy (FCS) provides an easy and reliable measure of these quantities. We modify existing schemes for performing FCS on a commercial confocal microscope to carry out these measurements, and provide an analysis routine that can yield the relevant quantities. Our method does not require any modification of the confocal microscope, yet it yields a robust measure of the resolution and sensitivity of the instrument. PMID- 15889425 TI - Detection of injected fluorescence-conjugated IgG in living mouse organs using "in vivo cryotechnique" with freeze-substitution. AB - In this experiment, we performed the "in vivo cryotechnique" in tandem with fluorescence microscopy. The fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit immunoglobulin (IgG) antibody (FITC-IgG) was directly injected into mouse livers or kidneys, which were then frozen in vivo by pouring an isopentane propane mixture (-193 degrees C) cooled in liquid nitrogen over these living organs. The organs were subsequently freeze-substituted in acetone containing paraformaldehyde at about -80 degrees C, then gradually brought up to a room temperature, infiltrated with 30% sucrose and refrozen. Some well-frozen areas 300-400 mum below the frozen tissue surface were cryocut into several slices. The slices were observed under the fluorescence microscope. By examining the distribution of FITC-IgG in the frozen livers, some aspects of functional blood circulation in the liver, such as the concept of the liver lobule, were reconfirmed. This also confirmed that the blood flow in the liver after the FITC IgG injection was normal. The subsequent preparation of the specimens with immunohistochemistry, using the tetramethylrhodamine (TRITC)-conjugated anti mouse IgG antibody, allowed us to visualize the localizations of both the original mouse IgG and the injected goat IgG in the cryosections with different color images. The experimental protocol presented demonstrates the in situ localization of the various proteins labeled with fluorescent probes, and it can, in conjunction with immunohistochemistry, localize proteins in cells and tissues. PMID- 15889426 TI - Occasional transsynaptic viral labeling in the central nervous system from the polycystic ovary induced by estradiol valerate. AB - Increased density of catecholaminergic nerves in the human polycystic ovary has been observed. The aim of the present study was to investigate the distribution of transsynaptically virus-labeled neurons in the central nervous system from the rat polycystic ovary to see whether is it different or not from that of cycling control rats. To induce a polycystic ovary, a single injection of estradiol valerate was given to adult female rats and 30 days later a neurotropic virus was injected into the right ovary. Rats were sacrificed 72 or 96 hours after viral infection. Weight of the ovaries of the estradiol valerate-treated rats was significantly lower compared to controls, and the histology of the ovaries of the treated rats displayed severely atretic large antral follicles. There was almost no viral labeling in the central nervous system from the ovaries showing precystic morphology, in spite of the fact that such altered organs are rich in nerve fibres. It is assumed that presently unidentified factors in the precystic ovary, presumably related to the link between the immune and the nervous system, might be involved in the infectivity of the virus, and thus be responsible for the lack of viral labeling from such an ovary. PMID- 15889427 TI - Imaging DNA molecules on mica surface by atomic force microscopy in air and in liquid. AB - DNA molecules immobilized on mica surface by various methods have been observed by atomic force microscopy both in air and in liquid. Divalent cations and 3 aminopropyltriethoxysilane (APTES) modified mica surface have been used to immobilize the DNA molecules. Optimal DNA and divalent cations concentration for AFM imaging are presented. Among the different methods of modifying mica surface with APTES, the water solution modifying method appears to get the best results. When using high DNA concentration for AFM imaging, DNA networks can be formed. A simple method to extend long DNA molecules is demonstrated. The optimal imaging conditions and AFM operating techniques are discussed. Different DNA immobilizing methods have been compared and evaluated. PMID- 15889428 TI - Videomicroscopy, image processing, and analysis of whole histologic sections of the human brain. AB - Serial histologic sections of a whole human brain may have extensions of up to 130 x 130 mm within the coronal plane around the temporal lobe. To date, however, technology has not provided a bright field microscope that is able to shift the object holder continuously in the x- and y-direction over such distances and still possess the same optical capabilities as comparable devices. We developed a new light microscope to continuously quantify such sections. We also developed the computing environment for controlling the device and for analyzing the data produced. In principle, we are now able to quantify each neuron of a human brain. The data ultimately will provide the most detailed structural information about the human brain ascertained thus far. Such detailed information of the spatial distribution of neurons is essential to develop realistic models for simulation of large-scale neuronal networks and to investigate the significance of neuronal arrangements with respect to neuronal signal processing in the CNS. After preprocessing of the data produced by the new microscope, we are able to detect lamination patterns in the spatial distribution of gravity centers of cells. Furthermore, morphological features like size of the projection area and mean staining intensity are visualized as a particle process. The particle process presents the sizes and staining intensity of perikaryons and allows a distinction of gray matter and white matter. These results provide evidence that the system works correctly and can be applied to a systematic analysis of a larger sequence of serial histologic sections. The objective of this study is to introduce the very large section analyzing microscope (VLSAM) and to present the initial data produced by the system. Moreover, we will discuss workload and future developments of the parallel image analysis system that are associated with the microscope. PMID- 15889429 TI - Maxillary sinus augmentation with Bio-Oss particles: a light, scanning, and transmission electron microscopy study in man. AB - Biological interactions occurring at the bone-biomaterial interface are critical for long-term clinical success. Bio-Oss is a deproteinized, sterilized bovine bone that has been extensively used in bone regeneration procedures. The aim of the present study was a comparative light, scanning, and electron microscopy evaluation of the interface between Bio-Oss and bone in specimens retrieved after sinus augmentation procedures. Under light microscopy, most of the particles were surrounded by newly formed bone, while in a few cases, at the interface of some particles it was possible to observe marrow spaces and biological fluids. Under scanning electron microscopy, in most cases, the particle perimeter appeared lined by bone that was tightly adherent to the biomaterial surface. Transmission electron microscopy showed that the bone tissue around the biomaterial showed all the phases of the bone healing process. In some areas, randomly organized collagen fibers were present, while in other areas, newly formed compact bone was present. In the first bone lamella collagen fibers contacting the Bio-Oss surface were oriented at 243.73 +/- 7.12 degrees (mean +/- SD), while in the rest of the lamella they were oriented at 288.05 +/- 4.86 degrees (mean +/- SD) with a statistically significant difference of 44.32 degrees (p < 0.001). In the same areas the intensity of gray value was 172.56 +/- 18.15 (mean +/- SD) near the biomaterial surface and 158.71 +/- 21.95 (mean +/- SD) in the other part of the lamella with an unstatistically significant difference of 13.79 (p = 0.071). At the bone-biomaterial interface there was also an electron-dense layer similar to cement lines. This layer had a variable morphology being, in some areas, a thin line, and in other areas, a thick irregular band. The analyses showed that Bio Oss particles do not interfere with the normal osseous healing process after sinus lift procedures and promote new bone formation. In conclusion, this study serves as a better understanding of the morphologic characteristics of Bio-Oss and its interaction with the surrounding tissues. PMID- 15889430 TI - Natriuretic peptides in ectopic myocardial tissues originating from mouse embryonic stem cells. AB - In a previous report we described the survival and contractile function of mouse embryonic stem cell-derived cardiomyocytes in the host retroperitoneum. To further understand the nature of embryonic stem cell-derived cardiomyocytes, the study assessed the synthesis of natriuretic peptides in ectopic myocardial tissues of embryonic stem cell origin. Cardiomyocytes formed in embryoid body outgrowths were transplanted into the retroperitoneum of adult nude mice, and the myocardial tissues that developed were characterized by RT-PCR and immunohistochemistry concerning atrial and brain natriuretic peptides (ANP, BNP). In the outgrowths of embryoid bodies in vitro, gene expression of ANP and BNP was detected by RT-PCR and granules positive for the peptides were identified in a few cardiomyocytes by light and electron microscopic immunocytochemistry. Seven days after transplantation the transplants exhibited multidifferentiated teratoma tissues. Developing chamber myocardial tissues positive for cardiac troponin I, cadherin, and connexin 43 were evident in the transplants, which contained ANP positive cardiomyocytes. Transplants with beating bundles were observed 30 days after transplantation, in which gene expression of both natriuretic peptides was detected. Myocardial tissues with abundant ANP-immunoreactivity, as well as with BNP-immunoreactivity to a lesser extent, were evident in the transplants. Also, myocardial tissues without immunoreactivity for natriuretic peptides were observed. Immunoelectron microscopy showed discernible secretory granules containing ANP and/or BNP in the cardiomyocytes. These results showed that part of the cardiomyocytes in embryonic stem cell-derived ectopic myocardial tissues are capable of producing natriuretic peptides, which suggests that they may be used as an endocrine source for cardiac hormones. PMID- 15889431 TI - No effect of a type I collagen gel coating in uncemented implant fixation. AB - Uncemented joint replacement with a variety of substrate materials, structures, and coatings are commonplace in arthroplasty. Even with specialized surgical preparation of bone, intimate contact between the implant and host bone may not always be achieved. This study evaluated the in vivo effect of fibrillar atelopeptide and PEG crosslinked collagens coatings placed directly into porous sintered bead structures on bone ingrowth using a skeletally mature bicortical, bilateral ovine tibia model. Bone ingrowth into the implants increased with time, although differences were not significant. At 4 weeks woven bone was present within the pores that remodeled with time. Significantly lower levels of ingrowth were observed in the intramedullary region of the implants when compared with the cortical region. Implant metal type did not affect ingrowth in both regions analyzed. Both fibrillar and crosslinked forms of dermal type I collagen did not significantly alter bone ingrowth. PMID- 15889432 TI - Incorporating hidden Markov models for identifying protein kinase-specific phosphorylation sites. AB - Protein phosphorylation, which is an important mechanism in posttranslational modification, affects essential cellular processes such as metabolism, cell signaling, differentiation, and membrane transportation. Proteins are phosphorylated by a variety of protein kinases. In this investigation, we develop a novel tool to computationally predict catalytic kinase-specific phosphorylation sites. The known phosphorylation sites from public domain data sources are categorized by their annotated protein kinases. Based on the concepts of profile Hidden Markov Models (HMM), computational models are trained from the kinase specific groups of phosphorylation sites. After evaluating the trained models, we select the model with highest accuracy in each kinase-specific group and provide a Web-based prediction tool for identifying protein phosphorylation sites. The main contribution here is that we have developed a kinase-specific phosphorylation site prediction tool with both high sensitivity and specificity. PMID- 15889433 TI - Elution study of unreacted Bis-GMA, TEGDMA, UDMA, and Bis-EMA from light-cured dental resins and resin composites using HPLC. AB - In the present work the elution of residual monomers from light-cured dental resins and resin composites into a 75% ethanol:water solution was studied using High-Performance Liquid Chromatography (HPLC). The resins studied were made by light-curing of bisphenol A glycol dimethacrylate (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA), urethane dimethacrylate (UDMA), ethoxylated bisphenol A glycol dimethacrylate [Bis-EMA(4)] and mixtures of these monomers. The resin composites were made from two commercial light-cured restorative materials (Z100 MP and Filtek Z250), the resin matrix of which is based on copolymers of these monomers. The effect of the curing time on the amount of monomers eluted was investigated. The concentration of the extractable monomers was determined at several immersion periods from 3 h to 30 days. For all the materials studied, it was observed that the chemical structure of the monomers used for the preparation of the resins, which defines the chemical and physical structure of the corresponding resin, directly affects the amount of eluted monomers, as well as the time needed for the elution of this amount. In the case of composites, it seems that the elution process it is not influenced by the presence of filler. PMID- 15889434 TI - Analysis of sterilization protocols for peptide-modified hydrogels. AB - Concerns about the efficacy of ethanol disinfection for implanted biomaterials prompted investigation of an alternative sterilization process, ultraviolet irradiation, for terminal sterilization of N-isopropylacrylamide-based hydrogels containing biomimetic peptides. Ultraviolet irradiation is more easily applied on a laboratory scale than gamma irradiation or electron beam, two commercially utilized methods; thus, UVC irradiation was investigated as a low-cost sterilization procedure that might be performed in laboratories prior to in vivo studies. UVC irradiation at 400 muW/cm(2) for up to 15 h did not prevent growth of Escherichia coli within the hydrogels, while ethanol disinfection did prevent growth for the duration of the experiment (120 h). Furthermore, UVC irradiation caused progressive degradation of peptides containing the Arg-Gly-Arg (RGD) domain. UVC irradiation cannot be used as a terminal sterilization process for peptide-modified materials. The system used in this study is not intended to be adequate for evaluating the sterility of medical devices in accordance with current Good Manufacturing Practice (cGMP); however, it remains a useful, low cost system for the preliminary evaluation of sterilization procedures in terms of their ability to eliminate pathogenic organisms while preserving the structure of biologically active molecules within in a laboratory setting. Ethanol treatment is still the preferred method for disinfection of bioactive materials containing peptides or UV-degradable groups. PMID- 15889435 TI - Use of the chicken lysozyme 5' matrix attachment region to generate high producer CHO cell lines. AB - Scaffold or matrix attachment region (S/MAR) genetic elements have previously been proposed to insulate transgenes from repressive effects linked to their site of integration within the host cell genome. We have evaluated their use in various stable transfection settings to increase the production of recombinant proteins such as monoclonal antibodies from Chinese hamster ovary (CHO) cell lines. Using the green fluorescent protein coding sequence, we show that S/MAR elements mediate a dual effect on the population of transfected cells. First, S/MAR elements almost fully abolish the occurrence of cell clones that express little transgene that may result from transgene integration in an unfavorable chromosomal environment. Second, they increase the overall expression of the transgene over the whole range of expression levels, allowing the detection of cells with significantly higher levels of transgene expression. An optimal setting was identified as the addition of a S/MAR element both in cis (on the transgene expression vector) and in trans (co-transfected on a separate plasmid). When used to express immunoglobulins, the S/MAR element enabled cell clones with high and stable levels of expression to be isolated following the analysis of a few cell lines generated without transgene amplification procedures. PMID- 15889436 TI - Bacterial colonization on different suture materials--a potential risk for intraoral dentoalveolar surgery. AB - In this in vivo and in vitro study on resorbable (Monocryl and nonresorbable (Deknalon) monofilament sutures used in intraoral dentoalveolar surgery the bacterial colonization was compared. For the in vivo study the sutures were applied in 11 patients during dental surgery. Eight days postoperative the sutures were removed and the adhered bacteria were isolated and identified by biochemistry, morphology, antibiotic susceptibility, and gas chromatography. The colonization was studied by scanning electron microscopy. Aerobic and anaerobic bacteria were isolated in nearly equal colony-forming units (cfu) on each suture. In comparison with Monocryl about 15% more aerobic and anaerobic strains were isolated on Deknalon. Regarding the pathogens only, about three times more anaerobic strains were isolated on both sutures in total. Additionally, more pathogens were found on Deknalon than on Monocryl (aerobic >40%, anaerobic >25%). The variety of bacteria correspond with purulent infections, not with normal oral flora. Intraindividual comparisons of cfu showed differences in dependence of the patient as described for subgingivale plaques. For the in vitro study the sutures were incubated with Streptococcus intermedius and Prevotella intermedia for 0.5 h. Scanning electron microscopy was performed to examine qualitatively the level of bacterial adherence. After 0.5 h the bacteria adhered very well. The colonization rate of Streptococcus intermedius on both sutures was similar. Coccoid bacteria within biofilms were seen. The growth of Prevotella intermedia was much better on Deknalon than on Monocryl. The risk of bacteremia at the time of suture removal is discussed. PMID- 15889437 TI - Collagen fiber orientation near dental implants in human bone: do their organization reflect differences in loading? AB - This study is the first to investigate the collagen fiber orientation in human bone around titanium dental implants after loading. Birefringence under circularly polarized light (CPL) and scanning electron microscopy (SEM) investigation were used to assess both collagen orientation and density. Twenty osseointegrated dental implants were retrieved from the mandible bone of 10 patients and were used in the present study. The implants were retrieved after 6 months of function. The birefringence measurements were performed on digitized images using both the peri-implant bone (PB) and the alveolar bone (AB) retrieved at the moment of implant placing. All images detected at 100x were measured using a software image analysis. The results showed that 32.96% (3.208 +/- 0.435 mm(2)) of the PB area was composed of transverse collagen fibers while only 19.70% (1.957 +/- 0.253 mm(2)) was composed of longitudinally collagen fibers. In the AB 26.99% (2.620 +/- 0.520 mm(2)) of the examined area was composed of transverse collagen fibers while 22.25% (2.160 +/- 0.320 mm(2)) was composed of longitudinally collagen fibers. The CPL measurements of the birefringence indicated that the difference in area between the two collagen fibers orientations was statistically significant (p = 0.03) for both AB and PB. In PB the transverse collagen fibers were significantly increased (p < 0.01) than in AB; also the longitudinal collagen fibers differ significantly in the two groups (p = 0.02). The transverse collagen fibers in PB are more present under the lower flank of the threads where the load acts with a compression vectors, while outside the tip of the implant threads the collagen fibers run more longitudinally due to the loads that act with a tensile vectors. SEM observations showed, in the first collagen layers facing the implant surface, a random direction of the collagen fibers, while in the areas away from the implant surface the collagen fibers were well oriented and run parallel. In conclusion, in these loaded dental implants, the bone adjacent to the first two implant threads shows a significant increase of the amount of transverse collagen fibers that were mainly associated with the lower flank of the threads where compressive load acts. The longitudinal collagen fibers also differs significantly from those found in the AB. PMID- 15889438 TI - Dose- and time-dependent effect of bioactive gel-glass ionic-dissolution products on human fetal osteoblast-specific gene expression. AB - Bioactive glasses dissolve upon immersion in culture medium, and release their constitutive ions into solution. There has been some evidence suggesting that these ionic-dissolution products influence osteoblast-specific processes. Here, the effect of 58S sol-gel-derived bioactive glass (60% SiO(2), 36% CaO, 4% P(2)O(5), in molar percentage) on primary osteoblasts derived from human fetal long bone explant cultures is investigated, and it is hypothesized that critical concentrations of sol-gel-dissolution products (consisting of a combination of simple inorganic ions) can enhance osteoblast phenotype in vitro by affecting the expression of a number of genes associated with the differentiation and extracellular matrix deposition processes. Cells were exposed to a range of 58S dosages continuously for a period of 4-14 days in monolayer cultures. Quantitative real-time RT-PCR analysis of a panel of osteoblast-specific markers showed a varied gene expression pattern in response to the material. The highest concentration of Ca and Si tested (96 and 50 ppm, respectively) promoted upregulation of gene expression for most markers (including alkaline phosphatase, osteocalcin, and osteopontin) at the latest time point, compared to non-58S treated control, although this observation was not statistically significant. The same 58S concentration produced higher ALP activity levels and increased proliferation throughout the culture period, compared to lower dosages tested; however, the results generated were again not statistically significant. The data overall suggest that no significant effect can be ascribed to the ionic products of 58S bioactive gel-glass dissolution tested here and their ability to stimulate osteoblastic marker gene expression. PMID- 15889440 TI - Current status of prosthetic bypass grafts: a review. AB - Polymers such as Dacron and polytetrafluoroethylene (PTFE) have been used in high flow states with relative success but with limited application at lower flow states. Newer polymers with greater compliance, biomimicry, and ability to evolve into hybrid prostheses, suitable as smaller vessels, are now being introduced. In view of the advances in tissue engineering, this makes possible the creation of an ideal off-the-shelf bypass graft. We present a broad overview of the current state of prosthetic bypass grafts. PMID- 15889446 TI - Teratology Society 45th Annual Meeting, June 25-30, 2005, St. Pete Beach, Florida, USA. Abstracts. PMID- 15889445 TI - Grafting reaction of poly(D,L)lactic acid with maleic anhydride and hexanediamine to introduce more reactive groups in its bulk. AB - Bioactivity of biomaterials was a new requirement, especially in tissue engineering and drug delivery. As a traditional used biomaterial, polylactide (PLA) had no bioactivity, of course, and it still had few reactive groups to introduce some bioactive molecules in its bulk. Here, we want to introduce carboxyl groups and amino groups in the side chain of PLA to get more reactive groups for incorporating bioactive molecular later and to maintain the structure of main chain to keep its biodegradability, and to settle the acidity of PLA during hydrolysis at the same time. It was performed as follows: first, maleic anhydride was covalently grafted onto the side chain of PLA by a free radical reaction at 100 degrees C for 20 h with BPO as the initiator. Then, by amidation with a maleic anhydride group on PLA at room temperature, hexanediamine was incorporated. The resulting polymers have been characterized via GPC, (13)C NMR, DSC, and TGA. The graft ratio was tested by titration. The pH changes during hydrolysis in 0.1 M PBS with pH 7.4 of PLA, MPLA, and HPLA were investigated. All the results showed that this research has grafted maleic anhydride and then hexanediamine in the bulk of PLA. The molecular weight degradation during reaction was less than 20%. The graft ratios of were 2.68, 2.36, and 1.86%, respectively in 5, 10, and 20% raw MA in MPLA; and the anhydride groups grafted in MPLA can completely react with hexanediamine at room temperature. The pH value of HPLA remained neutral within 12 weeks' hydrolysis compared with the resulted acidity of PLA and MPLA. PMID- 15889447 TI - Effect of myristoylated alanine-rich C kinase substrate (MARCKS) overexpression on hippocampus-dependent learning and hippocampal synaptic plasticity in MARCKS transgenic mice. AB - The myristoylated alanine-rich C kinase substrate (MARCKS) is a primary substrate of protein kinase C (PKC) thought to regulate membrane-filamentous actin cytoskeletal plasticity in response to PKC activity in the regulation of synaptic efficacy. We have recently reported that MARCKS expression is significantly elevated (45%) in the hippocampus of DBA/2J mice, which exhibit impaired hippocampus-dependent learning and hippocampal long-term potentiation (LTP), compared with C57BL/6J mice. The latter finding led us to hypothesize that elevations in MARCKS expression are detrimental to hippocampal plasticity and function. To assess this more directly, we examined hippocampal (CA1) paired pulse facilitation and LTP, and hippocampus-dependent learning in mice overexpressing MARCKS through the expression of a human MARCKS transgene (Tg+). The human MARCKS protein was confirmed to be expressed in the hippocampus of Tg+ mice but not in Tg- mice. Schaffer collateral paired-pulse facilitation, input output responses, and LTP did not differ between Tg+ and Tg- mice, indicating that neurotransmitter release, short-term, and long-term synaptic plasticity are not impaired by MARCKS overexpression. In the Morris water maze, Tg+ mice exhibited a mild but significant spatial learning impairment during initial acquisition, and a more severe impairment during reversal training. Tg+ did not exhibit impaired swim speed or visible platform performance relative to Tg- mice, indicating the absence of gross sensorimotor deficits. Fear conditioning to either context or cue was not impaired in Tg+ mice. Behavioral deficits could not be attributed to differences in hippocampal PKC isozyme (alpha beta(II), gamma, epsilon, zeta) or calmodulin expression, or alterations in hippocampal cytoarchitecture or infrapyramidal mossy fiber limb length. Collectively, these results indicate that elevations in MARCKS expression are detrimental to specific aspects of hippocampal function. PMID- 15889448 TI - Fifty years later: I. Michael Lerner's Genetic homeostasis (1954)--a valiant attempt to integrate genes, organisms and environment. PMID- 15889449 TI - Red cell mass and plasma volume measurements in polycythemia: evaluation of performance and practical utility. AB - BACKGROUND: Despite the absence of any systematic evidence for diagnostic utility, red cell mass (RCM) measurement has been endorsed as a major diagnostic criterion for polycythemia vera (PV) based on a set of eligibility criteria for a clinical trial formulated by an International PV Study Group in 1967. METHODS: A retrospective analysis was performed on a consecutive series of 105 patients who underwent blood volume measurements for evaluation of polycythemia. In a previous study, the authors had systematically compared RCM measurement by 51Cr-labeled erythrocytes and 125I-labeled human serum albumin and demonstrated equivalence between the two methods. In the current study, they used the latter method and applied the International Committee for Standardization in Haematology recommendations for result interpretation in order to evaluate test performance and practical utility. RESULTS: RCM exceeded the 98-99% limits of the reference range in 76%, 20%, 22%, and 57% of patients with PV (n = 25), secondary polycythemia (n = 35), spurious or apparent polycythemia (n = 38), and essential thrombocythemia (n = 7), respectively. Decreased plasma volume was rarely seen in any of the disease categories. In all instances of PV, the diagnosis was readily apparent, based on alternative clinical and laboratory tests, and did not require the additional information from blood volume measurement. Furthermore, alternative methods of PV diagnosis, based on disease-specific biological markers as well as bone marrow histology, are now available. CONCLUSIONS: The continued use of RCM and plasma volume measurements for the diagnosis of PV is no longer warranted. PMID- 15889450 TI - Socioeconomic status and the risk of cervical intraepithelial neoplasia grade 3 among oncogenic human papillomavirus DNA-positive women with equivocal or mildly abnormal cytology. AB - BACKGROUND: Low socioeconomic status (SES) is a reported risk factor for cervical carcinoma, but few studies have taken into account adequately the possibly confounding effects of oncogenic human papillomavirus (HPV) infection as well as access to screening and subsequent treatment. METHODS: Women (n = 5060 women) with a mean age of 27.5 years and with equivocal or mild cytologic cervical abnormalities were enrolled in the Atypical Squamous Cells of Undetermined Significance/Low-Grade Squamous Intraepithelial Lesion (ASCUS-LSIL) Triage Study (ALTS), a clinical trial that evaluated management strategies. The women were seen every 6 months for 2 years. The enrollment questionnaire assessed three indicators of SES: race/ethnicity, education, and source of payment for medical care. Multivariate logistic regression models were used to identify predictors of oncogenic HPV DNA positivity at enrollment and to assess associations between the SES indicators and risk of cervical intraepithelial neoplasia grade 3 (precancer) and carcinoma (> or = CIN3) identified throughout the study (n = 506 women) among oncogenic HPV-positive women (n = 3133 women). RESULTS: SES indicators were not associated significantly with oncogenic HPV infection after adjustment for age at enrollment, recent and lifetime number of sexual partners, study center, and smoking history. Among women with oncogenic HPV, the risk of > or = CIN3 increased with decreasing education (less than high school education: odds ratio [OR], 2.4; 95% confidence interval [95%CI], 1.5-3.7 vs. completed college). Black women (OR, 0.5; 95%CI, 0.4-0.7) and white/Hispanic women (OR, 0.4; 95%CI, 0.2 0.8) were at decreased risk for > or = CIN3 compared with white/non-Hispanic women. The source of payment for medical care was not associated with risk. CONCLUSIONS: Factors associated with lower SES, such as low education, may serve as a surrogate for unknown factors that influence progression to > or = CIN3 among women with oncogenic HPV infection. In this controlled setting with equalized follow-up and treatment, the decreased risk of > or = CIN3 associated with black and white/Hispanic race/ethnicity could be further examined. Ongoing efforts should emphasize methods for equalizing screening and follow-up among women of varying SES, regardless of race or ethnicity. PMID- 15889451 TI - Analysis of an interaction threshold in a mixture of drugs and/or chemicals. AB - Increasingly, humans are exposed to drug/chemical mixtures. These exposures can result from therapeutic interventions or environmental sources. Of interest is the interaction that may occur among the components of these mixtures. Since interaction can be dose-dependent, it is important to determine exposure levels to either exploit the benefits of the interaction in a therapeutic application or to avoid the effect of the interaction in the case of an environmental risk assessment. We propose generalized linear models that permit the estimation of interaction threshold boundaries. The methods developed are applied to the combination of ethanol and chloral hydrate. PMID- 15889452 TI - Significance testing for small microarray experiments. AB - Which significance test is carried out when the number of repeats is small in microarray experiments can dramatically influence the results. When in two sample comparisons both conditions have fewer than, say, five repeats traditional test statistics require extreme results, before a gene is considered statistically significant differentially expressed after a multiple comparisons correction. In the literature many approaches to circumvent this problem have been proposed. Some of these proposals use (empirical) Bayes arguments to moderate the variance estimates for individual genes. Other proposals try to stabilize these variance estimate by combining groups of genes or similar experiments. In this paper we compare several of these approaches, both on data sets where both experimental conditions are the same, and thus few statistically significant differentially expressed genes should be identified, and on experiments where both conditions do differ. This allows us to identify which approaches are most powerful without identifying many false positives. We conclude that after balancing the numbers of false positives and true positives an empirical Bayes approach and an approach which combines experiments perform best. Standard t-tests are inferior and offer almost no power when the sample size is small. PMID- 15889453 TI - A note on testing in Aalen's additive hazards regression models. AB - Aalen's additive hazards regression model is a useful alternative to the proportional hazards model for censored data regression. When used to compare treatments this approach leads to weighted comparisons of the crude estimate of the hazard rate of each group as compared to a baseline group. This is contrasted to the weighted log rank test from the proportional hazards model which compares each treatment's rate to the pooled rate. We show in this brief note that Aalen's suggestion for weights in this test leads to inconsistent tests in the sense that the test statistic depends on which group we pick for a baseline group. We show that 'consistent' tests are obtained by using common weight functions for all comparisons and we make some suggestions. PMID- 15889454 TI - Does it always help to adjust for misclassification of a binary outcome in logistic regression? AB - It is well known that in logistic regression, where the outcome is measured with error, a biased estimate of the association between the outcome and a risk factor may result if no proper adjustment is made. Hence, it seems tempting to always adjust for possible misclassification of the outcome. Here we show that it is not always beneficial to do so because, though the adjustment reduces the bias, it also inflates the variance, leading to a possibly larger mean squared error of the estimate. In the context of a data set on agricultural injuries, numerical evidence is provided through simulation studies. PMID- 15889456 TI - A case report of prenatal diagnosis of PentaX syndrome in association with isolated borderline ventriculomegaly. AB - We report of case of pentaX syndrome that was diagnosed prenatally. PentaX aneuploidy is very rare with only 25 cases reported so far. This is the third case of prenatal diagnosis of pentaX syndrome in 2nd trimester. This patients underwent an amniocentesis for ventriculomegaly which confirmed the diagnosis. PMID- 15889457 TI - Discontinuities in the DNA synthesized in an excision-defective strain of Escherichia coli following ultraviolet irradiation. 1968. PMID- 15889458 TI - Predicting the failure of 3 by 5. PMID- 15889459 TI - Prostitutes are people too. PMID- 15889460 TI - Ridding the world of infection. PMID- 15889461 TI - Doctors' group proposes one-child policy for India. PMID- 15889462 TI - The muddle of US electronic medical records. PMID- 15889463 TI - Guidelines set ethical bar for US stem cell research. PMID- 15889464 TI - Unethical clinical trials in Thailand: a community response. PMID- 15889465 TI - Zimbabwe's reversal of fortune. PMID- 15889466 TI - UN yields to pressure on needle-exchange programmes. PMID- 15889467 TI - Freeze-drying process design by manometric temperature measurement: design of a smart freeze-dryer. AB - PURPOSE: To develop a procedure based on manometric temperature measurement (MTM) and an expert system for good practices in freeze drying that will allow development of an optimized freeze-drying process during a single laboratory freeze-drying experiment. METHODS: Freeze drying was performed with a FTS Dura Stop/Dura-Top freeze dryer with the manometric temperature measurement software installed. Five percent solutions of glycine, sucrose, or mannitol with 2 ml to 4 ml fill in 5 ml vials were used, with all vials loaded on one shelf. Details of freezing, optimization of chamber pressure, target product temperature, and some aspects of secondary drying are determined by the expert system algorithms. MTM measurements were used to select the optimum shelf temperature, to determine drying end points, and to evaluate residual moisture content in real-time. MTM measurements were made at 1 hour or half-hour intervals during primary drying and secondary drying, with a data collection frequency of 4 points per second. The improved MTM equations were fit to pressure-time data generated by the MTM procedure using Microcal Origin software to obtain product temperature and dry layer resistance. Using heat and mass transfer theory, the MTM results were used to evaluate mass and heat transfer rates and to estimate the shelf temperature required to maintain the target product temperature. RESULTS: MTM product dry layer resistance is accurate until about two-thirds of total primary drying time is over, and the MTM product temperature is normally accurate almost to the end of primary drying provided that effective thermal shielding is used in the freeze drying process. The primary drying times can be accurately estimated from mass transfer rates calculated very early in the run, and we find the target product temperature can be achieved and maintained with only a few adjustments of shelf temperature. The freeze-dryer overload conditions can be estimated by calculation of heat/mass flow at the target product temperature. It was found that the MTM results serve as an excellent indicator of the end point of primary drying. Further, we find that the rate of water desorption during secondary drying may be accurately measured by a variation of the basic MTM procedure. Thus, both the end point of secondary drying and real-time residual moisture may be obtained during secondary drying. CONCLUSIONS: Manometric temperature measurement and the expert system for good practices in freeze drying does allow development of an optimized freeze-drying process during a single laboratory freeze-drying experiment. PMID- 15889468 TI - WHO: big gaps remain in global tuberculosis case detection. PMID- 15889469 TI - Is abstinence-only threatening Uganda's HIV success story? PMID- 15889470 TI - US Senate approves extra money for Global Fund. PMID- 15889471 TI - India's new patent bill threatens generic industry. PMID- 15889472 TI - Lymphogranuloma venereum spread linked to reporting delay. PMID- 15889473 TI - Avian influenza vaccine clinical trial begins in USA. PMID- 15889474 TI - Experiments shed light on early events in HIV infection. PMID- 15889475 TI - US response to anthrax scare "uncoordinated". PMID- 15889476 TI - New combination drug to treat malaria. PMID- 15889479 TI - Insulin independence after living-donor distal pancreatectomy and islet allotransplantation. AB - Rising demand for islet transplantation will lead to severe donor shortage in the near future, especially in countries where cadaveric organ donation is scarce. We undertook a successful transplantation of living-donor islets for unstable diabetes. The recipient was a 27-year-old woman who had had brittle, insulin dependent diabetes mellitus for 12 years. The donor, who was a healthy 56-year old woman and mother of the recipient, underwent a distal pancreatectomy. After isolation, 408 114 islet equivalents were transplanted immediately. The transplants functioned immediately and the recipient became insulin-independent 22 days after the operation. The donor had no complications and both women showed healthy glucose tolerance. Transplantation of living-donor islets from the distal pancreas can be sufficient to reverse brittle diabetes. PMID- 15889480 TI - [The 89th Congress of the Japanese Society of Legal Medicine. Kagawa, Japan. April 20-22, 2005. Abstracts]. PMID- 15889482 TI - Nursing science - working toward a healthier nation. Abstracts from the National Congress on the State of the Science Conference in Nursing Research. October 7-9, 2004. Washington, D.C., USA. PMID- 15889481 TI - [The 106th Congress of the Japanese Society of Otolaryngology. Osaka, Japan. May 19-21. 2005. Abstracts]. PMID- 15889483 TI - Mirror, mirror, on the wall . . . PMID- 15889485 TI - Exploring life by single-molecule fluorescence spectroscopy. Molecular characteristics hidden by ensemble experiments can be revealed by fluorescence. PMID- 15889486 TI - Medical image. Jaundice. PMID- 15889487 TI - Proceedings of the Institute of Electrical and Electronics Engineers (IEEE) Computer Society Bioinformatics Conference. August 14-16, 2002. Stanford, California, USA. PMID- 15889488 TI - Changing the look of voltammetry. Can FT revolutionize voltammetric techniques as it did for NMR? PMID- 15889489 TI - Systems biology: a boon for analytical chemists? PMID- 15889490 TI - 80 years of watching the evolutionary scenery. PMID- 15889491 TI - Proceedings of the annual conference of the European Macrophage and Dendritic Cell Society (EMDS). August 28-30, 2003. Leicester, United Kingdom. PMID- 15889492 TI - The use of injury surveillance databases to identify emerging injury hazards. AB - Among the many valuable uses of injury surveillance is the potential to alert health authorities and societies in general to emerging injury trends, facilitating earlier development of prevention measures. Other than road safety, to date, few attempts to forecast injury data have been made, although forecasts have been made of other public health issues. This may in part be due to the complex pattern of variance displayed by injury data. The profile of many injury types displays seasonality and diurnal variance, as well as stochastic variance. The authors undertook development of a simple model to forecast injury into the near term. In recognition of the large numbers of possible predictions, the variable nature of injury profiles and the diversity of dependent variables, it became apparent that manual forecasting was impractical. Therefore, it was decided to evaluate a commercially available forecasting software package for prediction accuracy against actual data for a set of predictions. Injury data for a 4-year period (1996 to 1999) were extracted from the Victorian Emergency Minimum Dataset and were used to develop forecasts for the year 2000, for which data was also held. The forecasts for 2000 were compared to the actual data for 2000 by independent t-tests, and the standard errors of the predictions were modelled by stepwise hierarchical multiple regression using the independent variables of the standard deviation, seasonality, mean monthly frequency and slope of the base data (R = 0.93, R(2) = 0.86, F(3, 27) = 55.2, p < 0.0001). Significant contributions to the model included the SD (beta = 1.60, p < 0.001), mean monthly frequency (beta = -0.72, p < 0.002), and the seasonality of the data (beta = 0.16, p < 0.02). It was concluded that injury data could be reliably forecast and that commercial software was adequate for the task. Variance in the data was found to be the most important determinant of prediction accuracy. Importantly, automated forecasting may provide a vehicle for identifying emerging trends. PMID- 15889493 TI - Abstracts of the 49th Meeting of the Italian Society for Biochemistry and Molecular Biology. Riccione, Italy, September 28-October 1, 2004. PMID- 15889494 TI - More negative results for vitamin E. PMID- 15889495 TI - Sirolimus does double duty after organ transplantation. PMID- 15889497 TI - Radiotherapy versus surgery for prostate cancer. PMID- 15889499 TI - Diffusion imaging to assess brain-cancer treatment. PMID- 15889500 TI - Is lymphangiogenesis as important as angiogenesis? PMID- 15889502 TI - Bevacizumab and chemotherapy improves survival in NSCLC. PMID- 15889503 TI - Malaria gene linked to prostate-cancer incidence. PMID- 15889504 TI - Abstracts of the 10th Annual Interventional Vascular Therapeutics Angioplasty Summit-Transcatheter Cardiovascular Therapeutics Asia Pacific Symposium. April 28 30, 2005, Seoul, Korea. PMID- 15889506 TI - Diabetes and Ageing. Abstracts of the 11th Annual International Conference of the UDOP, WHO/PAHO, CFNI. March 3-6, 2005, Kingston, Jamaica. PMID- 15889505 TI - Digestive Disease Week, American Society for Gastrointestinal Endoscopy meeting abstracts. May 2005, Chicago, Illinois, USA. PMID- 15889507 TI - Assisted living: a regulation dilemma. Improving assisted living is no easy job, lawmakers have to look at quality, economy and affordability. PMID- 15889508 TI - Asylum-seekers and national histories of detention. PMID- 15889509 TI - Sheep farming in Sutherland in the eighteenth century. PMID- 15889510 TI - Acupuncture for treatment of climacteric syndrome--a report of 35 cases. AB - OBJECTIVE: To evaluate the effects of acupuncture for treatment of climacteric syndrome. METHODS: The 65 cases in this series were randomly assigned to the acupuncture group (N=35) and the control group (N=30), with the former treated by acupuncture and the latter by medication respectively. The therapeutic effects were evaluated by means of the clinical outcome, improvement in the symptom scores and the results of radioimmunoassays. RESULTS: In the acupuncture group, 12 cases were cured, 16 cases markedly effective, and 6 cases improved, the total effective rate being as high as 97.14%. The decrease in the symptom scores, and especially the elevation of the decreased E2 level and the decrease of the increased FSH and LH levels, demonstrated that acupuncture therapy was superior to medication. CONCLUSION: Acupuncture for regulating the mental activities and reinforcing the kidney is an effective therapy for climacteric syndrome. PMID- 15889511 TI - An analysis on the therapeutic effects of auriculo-acupuncture in 38 obstinate hiccup cases of different races. PMID- 15889512 TI - Experience in the clinical application of Naokong (GB19). PMID- 15889513 TI - Electro-acupuncture treatment for the upper segment ureterolithiasis under B ultrasonography. AB - In order to explore the effects of the local strong stimulation generated by electro-acupuncture for treatment of the upper segment ureterolithiasis, a controlled study was carried out among the treatment group (electro-acupuncture with strong stimulation), the control group I (medication) and the control group II (conventional acupuncture). The results showed that the differences in the cure rate and the total effective rate between the treatment group and the two control groups were significant in statistical analysis (P<0.05 and P<0.01 respectively). This indicates that better therapeutic effects can be obtained by the local strong stimulation generated by electro-acupuncture for treatment of the upper segment ureterolithiasis. PMID- 15889514 TI - Treatment of chronic pelvic inflammation with acupuncture and TDP irradiation--a report of 23 cases. PMID- 15889515 TI - Chinese herbal medicine and acupuncture used for rehabilitation of the impaired urinary system in patients with paraplegia--a report of 52 cases. PMID- 15889516 TI - Point injection of injectio radici astragali for treatment of post-chemotherapy adverse reactions. PMID- 15889517 TI - Treatment of peptic ulcer with xiao jianzhong tang--a report of 80 cases. PMID- 15889518 TI - The therapeutic effects of ling gui zhu gan tang mixture in 50 psychotic patients with obesity induced by the psychoactive drugs. PMID- 15889519 TI - A clinical study on haunglian fire-purging mixture in treatment of 46 cases of primary hypertension. AB - In order to observe the therapeutic effects of Huanglian Fire-Purging Mixture ([symbol: see text]) on primary hypertension, 46 cases of primary hypertension in the treatment group were treated with Huanglian Fire-Purging Mixture to clear away heat from the liver, relieve mental stress, purge fire and remove toxin; and the other 26 cases of primary hypertension in the control group were treated with Niuhuang Bolus for Lowering Blood Pressure ([symbol: see text]). The effect in the treatment group was obviously superior to that in the control group (P<0.05). The Huanglian Fire-Purging Mixture shows noticeable effects 3-6 hours after medication. The mixture can improve the clinical symptoms, the left ventricular diastolic function and myocardial ischemia, correct dyslipoproteinemia and dysglycemia, and reduce blood viscosity. And it is safe and with no obvious adverse reactions. PMID- 15889520 TI - TCM treatment of thromboangiitis obliterans--a report of 64 cases. PMID- 15889521 TI - TCM differential treatment of IgA nephrosis. PMID- 15889522 TI - Dr. Zang Kuntang's experience in treating diabetes. PMID- 15889523 TI - TCM treatment for 63 cases of senile dyssomnia. AB - In order to study the therapeutic effects of the TCM drugs on senile dyssomnia, 121 such patients were randomly divided into a treatment group of 63 cases (given the TCM drugs) and a control group of 58 cases (given estazolam). The changes shown in the SDRS and HAMA scores and the other indexes were observed in both of the two groups to evaluate the therapeutic effects. The results showed that the effective rate was 76.3% in the treatment group, and it was 69.1% in the control group; and that the TCM drugs had better effects in improving such symptoms as lethargy, dry mouth, and rebound of insomnia. Therefore, it can be concluded that the effect of the TCM drugs isbetter for senile dyssomnia than that of the Western drug estazolam. PMID- 15889525 TI - Clinical records for acupuncture treatment of migraine. PMID- 15889524 TI - Effects of shenmai injection on blood SOD activity and MDA level in senile patients with coronary heart disease. AB - In order to approach the effects of Shenmai Injection on SOD activity and MDA level in the senile patients with coronary heart disease, 48 cases, who had stenosis of over 70% in more than one branches of the coronary arteries, were assigned randomly into a treatment group (given Shenmai Injection plus the routine treatment) and a control group (given the routine treatment only). The superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were determined before treatment and at the end of a 3-week treatment. The results showed that in the treatment group, the SOD activity was significantly increased (P<0.05) and the MDA level markedly decreased (P<0.01) in the treatment group. It can be concluded that Shenmai Injection may enhancethe antioxidant ability of the senile patients with coronary heart disease. PMID- 15889526 TI - Regulative actions of the Chinese drugs for tonifying the kidney on gene expression of the hypothalamic GnRH, pituitary FSH, LH and osteoblastic BGP. AB - It is found that the drugs for nourishing yin to reduce pathogenic fire can significantly down-regulate, and the drugs for tonifying the kidney to replenish essence can up-regulate mRNA expression of the hypothalamic GnRH, pituitary FSH, LH and osteoblastic BGP, indicating that the Chinese drugs for tonifying the kidney can regulate gene expression of the hypothalamic GnRH, pituitary FSH, LH, and osteoblastic BGP, which is possibly one of the main mechanisms of the Chinese drug for tonifying the kidney, regulating ephebic development process andimproving skeletal development in sexual precocity children. PMID- 15889527 TI - Effects of electroacupuncture on the contents of NO, ET and T-AOC in the brain tissues of the cerebral hemorrhage model rats. AB - In the cerebral hemorrhage model rats established by injection of collagenase and heparin into caudate nucleus, the effects of electroacupuncture (EA) on the contents of nitric oxide (NO) and endothelin (ET), and total anti-oxidation capability (T-AOC) in the brain tissues were investigated. It is found that the content of NO in the Shuigou EA group lowered, ET decreased and the T-AOC raised significantly in both the Fengfu EA group and the Shuigou EA group (all P<0.05) as compared with the model group, indicating that acupuncture can reduce the contents of ET and NO, and increase the T-AOC in the brain tissues of the rats with cerebral hemorrhage. PMID- 15889528 TI - Protective action of acupuncture and moxibustion on gastric mucosa in model rats with chronic atrophic gastritis. AB - OBJECTIVE: To probe the mechanism of acupuncture and moxibustion in atrophic gastritis so as to provide a basis for clinical treatment. METHOD: Observe the effects of acupuncture and moxibustion at the points of Zusanli, Zhongwan and Tianshu on gastric mucosa in model rats with chronic atrophic gastritis. RESULTS: Acupuncture and moxibustion can increase the contents of PGE2alpha, PGF2alpha and cAMP, and decrease the content of cGMP in the tissue of gastric mucosa. CONCLUSION: Acupuncture and moxibustion shows cytoprotection on gastric mucosa, so it is an effective method for treating chronic atrophic gastritis. PMID- 15889529 TI - Recent advances in TCM treatment of diabetic gangrene. PMID- 15889530 TI - Acupuncture treatment of menopausal syndrome. PMID- 15889531 TI - Host cell targets in HCV therapy: novel strategy or proven practice? AB - The development of novel antiviral drugs against hepatitis C is a challenging and competitive area of research. Progress of this research has been hampered due to the quasispecies nature of the hepatitis C virus, the absence of cellular infection models and the lack of easily accessible and highly representative animal models. The current combination therapy consisting of interferon-alpha and ribavirin mainly acts by supporting host cell defence. These therapeutics are the prototypic representatives of indirect antiviral agents as they act on cellular targets. However, the therapy is not a cure, when considered from the long-term perspective, for almost half of the chronically infected patients. This draws attention to the urgent need for more efficient treatments. Novel anti-hepatitis C treatments under study are directed against a number of so-called direct antiviral targets such as polymerases and proteases, which are encoded by the virus. Although such direct antiviral approaches have proven to be successful in several viral indications, there is a risk of resistant viruses developing. In order to avoid resistance, the development of indirect antiviral compounds has to be intensified. These act on host cell targets either by boosting the immune response or by blocking the virus host cell interaction. A particularly interesting approach is the development of inhibitors that interfere with signal transduction, such as protein kinase inhibitors. The purpose of this review is to stress the importance of developing indirect antiviral agents that act on host cell targets. In doing so, a large source of potential targets and mechanisms can be exploited, thus increasing the likelihood of success. Ultimately, combination therapies consisting of drugs against direct and indirect viral targets will most probably provide the solution to fighting and eradicating hepatitis C virus in patients. PMID- 15889532 TI - Oligonucleotides and polyribonucleotides: a review of antiviral activity. AB - Current antiviral therapies are insufficient for treating emerging, re-emerging and established viral diseases. In an effort to find new therapeutics, oligo- and polyribonucleotides are being studied for their antiviral capabilities. Studies have shown that uniquely modified single- and double-stranded nucleic acid constructs are effective in inhibiting viral proliferation by various mechanisms. This review gives a brief history and highlights the development of oligo- and polyribonucleotides as antiviral agents primarily in the fields of interferon induction, mRNA complementation and reverse transcriptase inhibition. PMID- 15889533 TI - HIV-1 subtype C drug-resistance background among ARV-naive adults in Botswana. AB - Current HIV-1 antiretroviral (ARV) drug resistance knowledge is limited to HIV-1 subtype B (HIV-1B). We addressed whether unique genetic and phenotypic properties of HIV-1 subtype C (HIV-1C), southern Africa's most prevalent subtype, may foment earlier and/or distinct resistance mutations. Population-level HIV-1C genotypes were evaluated with respect to drug resistance prevalence before Botswana's public ARV treatment programme began. Viruses were genotyped from 11 representative districts of northern and southern Botswana, and consensus sequences from these 71 individuals and 51 previously reported sequences from HIV positive blood donors were constructed. Phylogenetic analysis classified all 71 sequences but one, which exhibited pol gene mosaicism, as HIV-1C. The protease and reverse transcriptase coding region had no detectable known primary mutations associated with HIV-1B protease inhibitor (PI) drug resistance. Secondary mutations associated with PI drug resistance were found in all sequences. Several HIV-1C-specific polymorphic sites were found across the pol gene. Northern and southern Botswana viral sequences showed no significant differences from each other. Population genotyping shows that, without countrywide ARV treatment, HIV 1C-infected Batswana harbour virtually no primary mutations known to confer resistance to the three major HIV-1B ARV drug classes. Some secondary PI mutations and polymorphic sites in the protease enzyme necessitate continuous population monitoring, particularly after introduction of countrywide ARV treatment in Botswana. Although its PI resistance development rate and kinetics are not known, our data may suggest increased susceptibility and readiness of HIV 1C to develop resistance under drug pressure when the PI class of drugs is used. PMID- 15889534 TI - Comparative pharmacokinetics of Racivir, (+/-)-beta-2',3'-dideoxy-5-fluoro-3' thiacytidine in rats, rabbits, dogs, monkeys and HIV-infected humans. AB - Racivir is a 50:50 racemic mixture of the (-)- and (+)-beta-enantiomers of 2' deoxy-3'-thia-5-fluorocytosine (FTC), which is being developed for the treatment of HIV and hepatitis B virus (HBV). The (+)-enantiomer of FTC is approximately 10 20-fold less potent than (-)-FTC, but it selects for a different HIV mutation in human lymphocytes. Plasma concentrations from a group of 54 rats, 12 pregnant rabbits and 60 dogs enrolled in large toxicity studies using a wide variety of oral doses, were compared using non-compartment pharmacokinetic modelling versus dose, treatment duration, species and gender. The pharmacokinetics of Racivir were also compared with those of a previously published pharmacokinetic study in rhesus monkeys and with data from HIV-infected human male volunteers. The (+) FTC, but not the (-)-enantiomer, can be deaminated to the non-toxic inactive metabolite (+)-FTU. Therefore, the plasma exposure to (+)-FTU was also determined. The order of relative plasma exposure to (+)-FTU was rhesus monkeys > humans > pregnant rabbits > dogs > rats. Allometric scaling was performed to relate systemic clearance/fraction of drug absorbed (Cl/F) and terminal phase volume of distribution (Vbeta/F) versus species body weights. No individual animal species mimicked the Cl/F values in humans. However, allometric scaling using a combination of rats, pregnant rabbits and monkeys predicted the mean human Cl/F value better than a combination of rats and rabbits only (within 0.24 and SD of mean vs 0.81 SD of the observed mean value). Similarly, human Vbeta/F values were best predicted using a combination of rat and monkey data (within 0.64 SD of mean value). Species demonstrating greater deamination to (+)-FTU tended to have greater than predicted Cl/F values. The Cmax values of dogs were the closest to humans, but were statistically different. This study highlights the importance of selecting animal species that demonstrate similar cytidine deaminase activity to humans when performing preclinical dosing studies on Racivir and other antiviral agents that are substrates for mammalian cytidine deaminases. PMID- 15889535 TI - A throat lozenge containing amyl meta cresol and dichlorobenzyl alcohol has a direct virucidal effect on respiratory syncytial virus, influenza A and SARS-CoV. AB - A potent virucidal mixture containing amyl metacresol and dichlorobenzyl alcohol at low pH inactivated enveloped respiratory viruses influenza A, respiratory synctial virus (RSV) and severe acute respiratory syndrome coronavirus (SARS-CoV) but not viruses with icosahedral symmetry, such as adenoviruses or rhinoviruses. A titre of approximately 3.5 log10 TCID50 was reduced to below the level of detection within two minutes. Electron microscopy of purified influenza A virus showed extensive clumping and morphological changes in spike configuration after contact with the virucidal mixture, but no overt destruction of the viral membrane. We conclude that, formulated as a lozenge, the mixture could have significant effects in reducing the infectivity of certain infectious viruses in the throat and presumably in cough droplets, thus reducing, theoretically, opportunities for person-to-person transmission. PMID- 15889536 TI - Geldanamycin, a potent and specific inhibitor of Hsp90, inhibits gene expression and replication of human cytomegalovirus. AB - The effect of geldanamycin (GA), a specific inhibitor of heat shock protein 90 (Hsp90), on gene expression and replication of human cytomegalovirus (HCMV) was studied in human embryonic lung (HEL) fibroblasts. Kinetic analysis indicated that GA delayed synthesis of major immediate early (MIE), early and late viral proteins, and blocked a second tier of the synthesis of these proteins that occurred in untreated cells after 48 h post-infection (pi). Moreover, when HCMV infected HEL cells were maintained with medium containing 40 nM GA for 6 days, with medium changes at 2-day intervals, the virus yield was reduced to an undetectable level. On a molecular level, the cellular kinase Akt and the transcription factor NFkappaB were activated in HCMV-infected cells within 30 min pi. NFkappaB was shown to be essential for MIE gene expression. However, in GA treated cells, activation of both Akt and NFkappaB was greatly inhibited. Because LY294002, an inhibitor of cellular phosphatidylinositol 3-kinase (PI3-K), also prohibited HCMV-mediated activation of Akt and NFkappaB and synthesis of the MIE proteins, PI3-K signalling was necessary for expressing the MIE genes. These results suggest that the inhibitory effect of GA on HCMV replication is primarily caused by the disruption of the PI3-K signalling pathway, leading to the activation of NFkappaB, which plays a crucial role in expression of the critical MIE genes. PMID- 15889537 TI - [Immunomodulation therapy for autoimmune diseases. New hope for patients with autoimmune diseases]. AB - The number of patients suffering from autoimmune diseases is increasing and the prognosis of the patients is improving. The article gives an overview of the present status of immunomodulatory therapies. The author deals with the non specific and the selective forms of immunosuppression. He gives a short summary about biological treatments, the induction of tolerability and about anti cytokine therapies, In this review the modern treatment strategies of rheumatoid arthritis are also discussed, among them the early, the combined, the antigen specific and the stem cell treatment modalities are detailed. It is emphasised that the individual differences are important to take into consideration in the treatment of autoimmune diseases. PMID- 15889538 TI - [The role of aldosterone-antagonists in the treatment of congestive heart failure]. AB - The role of aldosterone-antagonists in the treatment of congestive heart failure. Despite the advances of the treatment of congestive heart failure, nearly half of the patients diagnosed with this disease five years ago are alive today. Experimental and human studies have demonstrated, that under special pathologic condition, the heart extracts aldosterone, and aldosterone extraction in the heart stimulates increased collagen turnover culminating in ventricular remodeling. Aldosterone blockade has been shown to be effective in reducing total mortality and hospitalization for heart failure in patients with systolic left ventricular dysfunction due to chronic heart failure (RALES study with spironolactone) and in patients with systolic left ventricular dysfunction post acute myocardial infarction (EPHESUS study with eplerenone). These clinical studies have shown that mineralocorticoid receptor activation remains important despite the use of angiotensin converting enzyme inhibitor or angiotensin receptor blocking agent and a beta blocker. In the ACC/AHA (and in the European and Hungarian) guidelines for the evolution and management of chronic heart failure, the indication of spironolactone was defined of Class Ila, Level of Evidence: B in CHF of stage C. The eplerenone (in US: INSPRA) was approved for the management of CHF patients after myocardial infarction with ejection fraction < 40%. Eplerenone, compared with spironolactone, is associated with a lower incidence of gynecomastia and other sex hormone-related adverse effect (breast pain, menstrual abnormalities). The spironolactone should not be used in patients with a creatinine above 220 mikromol/l. Despite the guidelines recommendation, spironolactone has been widely used in patients without consideration of their functional class or ejection fraction, without optimization of background treatment with ACE inhibitors and beta-blockers. PMID- 15889539 TI - [The plasma homocysteine, folic acid and vitamin B12 levels in young people with high risk for cardiovascular disease and its relation to metylentetrahydrofolate reductase (MTHFR) gene polymorphism]. AB - INTRODUCTION: Hyperhomocysteinemia is an independent risk factor for cardiovascular morbidity. AIM: The study was designed to evaluate the total homocysteine level and MTHFR C677T polymorphism frequency of 122, healthy, young adults who had increased risk for cardiovascular disease. The serum levels of folic acid and vitamin B12 were also measured. METHODS: Immunoassay, PCR-RFLP methods were used. The statistical analysis was performed by SPSS program. RESULTS: The frequency of the gene-polymorphism was not different significantly in the study group compared to a Hungarian neonatal sample: although in the increased risk group the frequency of homozygous 677TT polymorphism was higher (14.8%), and heterozygosity was smaller (41%). There was no association between MTHFR gene polymorphism and homocysteine levels. A significant negative correlation was found between the folic acid and homocysteine, and between the vitamin B12 and homocysteine levels correlating with the literature. The mean serum total homocysteine level of the group without vitamin supplementation (n: 86) was 9.8 +/- 3.3 micromol/l, while in the other group with vitamin uptake (n: 36) this level was 7.5 +/- 3.0 micromol/l. There was a significant difference between the homocysteine levels of men and women. CONCLUSION: The results of the study correlate with the literature. It would be useful to call the attention of the Hungarian population to the importance of vitamin supply. PMID- 15889540 TI - [Adult renal neoplasms in the material of the Pathology Department of the Szeged University]. AB - AIMS: The authors investigated the frequencies of the various histological types of adult renal tumours. METHODS: The slides of 469 nephrectomies performed in the Department of Urology, Szeged University between 1990 and 2003 were revised according to the 1997 Heidelberg and 2004 WHO classification schemes. RESULTS: 86.7% of all the tumours (n = 407) were malignant. Among the malignant tumours, the frequency of renal cell carcinomas was 91.1% (n = 371). 88.4% of the renal cell carcinomas (n = 328) were of conventional type, 5.6% (n = 21) were papillary and 4% (n = 15) were chromophobe. The authors observed 3 Bellini duct, 1 mucinous tubular and 3 non-classifiable carcinomas, with a combined incidence of 1.8%. 84.5% of the conventional carcinomas were clear cell (n = 277), 8.8% were eosinophilic granular (n = 29), 3.9% were multilocular cystic (n = 13) and 2.7% were sarcomatoid carcinomas (n = 9). The median age of the patients with conventional carcinoma was 60 (median, range: 25-84), in the papillary group it was 62 (43-78), and in the chromophobe group was 59 (17-77). The median age of patients affected by transitional cell carcinoma was 64 (range: 45-81). As far as benign tumours are concerned (13.2%, n = 62), oncocytomas (n = 37, 7.8% of all the tumours) affected mainly females, whereas angiomyolipomas (n = 21, 4.4% of all the tumours) occurred in females only. In 13 oncocytoma cases, the tumours were initially diagnosed as malignant. CONCLUSIONS: Adult malignant renal tumours affect mainly patients around the age of 60. The commonest diagnosis was clear cell carcinoma of conventional type. The incidence of clear cell carcinoma was 5% higher than that reported in the literature (84.5% vs 70-80%) whereas that of papillary carcinoma was 5% lower (5% vs 10-15%). In comparison with the literature data, oncocytomas were relatively common (8% instead of 3%), and not rarely, it was difficult to distinguish them from renal cell carcinomas. PMID- 15889541 TI - [Treatment of sleep disorders]. PMID- 15889542 TI - [Correspondence of Santiago Ramon y Cajal and Dezso Miskolczy between 1925 and 1936]. PMID- 15889543 TI - [Robert Martyn M.D. (1904-1999)]. PMID- 15889544 TI - Interplay between intensity standardization and inhomogeneity correction in MR image processing. AB - Image intensity standardization is a postprocessing method designed for correcting acquisition-to-acquisition signal intensity variations (nonstandardness) inherent in magnetic resonance (MR) images. Inhomogeneity correction is a process used to suppress the low frequency background nonuniformities (inhomogeneities) of the image domain that exist in MR images. Both these procedures have important implications in MR image analysis. The effects of these postprocessing operations on improvement of image quality in isolation has been well documented. However, the combined effects of these two processes on MR images and how the processes influence each other have not been studied thus far. In this paper, we evaluate the effect of inhomogeneity correction followed by standardization and vice-versa on MR images in order to determine the best sequence to follow for enhancing image quality. We conducted experiments on several clinical and phantom data sets (nearly 4000 three dimensional MR images were analyzed) corresponding to four different MRI protocols. Different levels of artificial nonstandardness, and different models and levels of artificial background inhomogeneity were used in these experiments. Our results indicate that improved standardization can be achieved by preceding it with inhomogeneity correction. There is no statistically significant difference in image quality obtained between the results of standardization followed by correction and that of correction followed by standardization from the perspective of inhomogeneity correction. The correction operation is found to bias the effect of standardization. We demonstrate this bias both qualitatively and quantitatively by using two different methods of inhomogeneity correction. We also show that this bias in standardization is independent of the specific inhomogeneity correction method used. The effect of this bias due to correction was also seen in magnetization transfer ratio (MTR) images, which are naturally endowed with the standardness property. Standardization, on the other hand, does not seem to influence the correction operation. It is also found that longer sequences of repeated correction and standardization operations do not considerably improve image quality. These results were found to hold for the clinical and the phantom data sets, for different MRI protocols, for different levels of artificial nonstandardness, for different models and levels of artificial inhomogeneity, for different correction methods, and for images that were endowed with inherent standardness as well as for those that were standardized by using the intensity standardization method. Overall, we conclude that inhomogeneity correction followed by intensity standardization is the best sequence to follow from the perspective of both image quality and computational efficiency. PMID- 15889545 TI - Improving the forward solver for the complete electrode model in EIT using algebraic multigrid. AB - Image reconstruction in electrical impedance tomography is an ill-posed nonlinear inverse problem. Linearization techniques are widely used and require the repeated solution of a linear forward problem. To account correctly for the presence of electrodes and contact impedances, the so-called complete electrode model is applied. Implementing a standard finite element method for this particular forward problem yields a linear system that is symmetric and positive definite and solvable via the conjugate gradient method. However, preconditioners are essential for efficient convergence. Preconditioners based on incomplete factorization methods are commonly used but their performance depends on user tuned parameters. To avoid this deficiency, we apply black-box algebraic multigrid, using standard commercial and freely available software. The suggested solution scheme dramatically reduces the time cost of solving the forward problem. Numerical results are presented using an anatomically detailed model of the human head. PMID- 15889546 TI - Automatic detection of red lesions in digital color fundus photographs. AB - The robust detection of red lesions in digital color fundus photographs is a critical step in the development of automated screening systems for diabetic retinopathy. In this paper, a novel red lesion detection method is presented based on a hybrid approach, combining prior works by Spencer et al. (1996) and Frame et al. (1998) with two important new contributions. The first contribution is a new red lesion candidate detection system based on pixel classification. Using this technique, vasculature and red lesions are separated from the background of the image. After removal of the connected vasculature the remaining objects are considered possible red lesions. Second, an extensive number of new features are added to those proposed by Spencer-Frame. The detected candidate objects are classified using all features and a k-nearest neighbor classifier. An extensive evaluation was performed on a test set composed of images representative of those normally found in a screening set. When determining whether an image contains red lesions the system achieves a sensitivity of 100% at a specificity of 87%. The method is compared with several different automatic systems and is shown to outperform them all. Performance is close to that of a human expert examining the images for the presence of red lesions. PMID- 15889547 TI - STACS: new active contour scheme for cardiac MR image segmentation. AB - The paper presents a novel stochastic active contour scheme (STACS) for automatic image segmentation designed to overcome some of the unique challenges in cardiac MR images such as problems with low contrast, papillary muscles, and turbulent blood flow. STACS minimizes an energy functional that combines stochastic region based and edge-based information with shape priors of the heart and local properties of the contour. The minimization algorithm solves, by the level set method, the Euler-Lagrange equation that describes the contour evolution. STACS includes an annealing schedule that balances dynamically the weight of the different terms in the energy functional. Three particularly attractive features of STACS are: 1) ability to segment images with low texture contrast by modeling stochastically the image textures; 2) robustness to initial contour and noise because of the utilization of both edge and region-based information; 3) ability to segment the heart from the chest wall and the undesired papillary muscles due to inclusion of heart shape priors. Application of STACS to a set of 48 real cardiac MR images shows that it can successfully segment the heart from its surroundings such as the chest wall and the heart structures (the left and right ventricles and the epicardium.) We compare STACS' automatically generated contours with manually-traced contours, or the "gold standard," using both area and edge similarity measures. This assessment demonstrates very good and consistent segmentation performance of STACS. PMID- 15889548 TI - Generalized likelihood ratio tests for complex fMRI data: a simulation study. AB - Statistical tests developed for the analysis of (intrinsically complex valued) functional magnetic resonance time series, are generally applied to the data's magnitude components. However, during the past five years, new tests were developed that incorporate the complex nature of fMRI data. In particular, a generalized likelihood ratio test (GLRT) was proposed based on a constant phase model. In this work, we evaluate the sensitivity of GLRTs for complex data to small misspecifications of the phase model by means of simulation experiments. It is argued that, in practical situations, GLRTs based on magnitude data are likely to perform better compared to GLRTs based on complex data in terms of detection rate and constant false alarm rate properties. PMID- 15889549 TI - Guide wire reconstruction and visualization in 3DRA using monoplane fluoroscopic imaging. AB - A method has been developed that, based on the guide wire position in monoplane fluoroscopic images, visualizes the approximate guide wire position in the three dimensional (3-D) vasculature, that is obtained prior to the intervention with 3 D rotational X-ray angiography (3DRA). The method assumes the position of the guide wire in the fluoroscopic images is known. A two-dimensional feature image is determined from the 3DRA data. In this feature image, the guide wire position is determined in a two-step approach: a mincost algorithm is used to determine a suitable position for the guide wire, and subsequently a snake optimization technique is applied to move the guide wire to a better position. The resulting guide wire can then be visualized in 3-D in combination with the 3DRA dataset. The reconstruction accuracy of the method has been evaluated using a 3DRA image of a vascular phantom filled with contrast, and monoplane fluoroscopic images of the same phantom without contrast and with a guide wire inserted. The evaluation has been performed for different projection angles, and with different parameters for the method. The final result does not appear to be very sensitive to the parameters of the method. The average mean error of the estimated 3-D guide wire position is 1.5 mm, and the average tip distance is 2.3 mm. The effect of inaccurate C-arm geometry information is also investigated. Small errors in geometry information (up to 1 degrees) will slightly decrease the 3-D reconstruction accuracies, with an error of at most 1 mm. The feasibility of this approach on clinical data is demonstrated. PMID- 15889550 TI - Precise robot-assisted guide positioning for distal locking of intramedullary nails. AB - This paper presents a novel image-guided robot-based system to assist orthopedic surgeons in performing distal locking of long bone intramedullary nails. The system consists of a bone-mounted miniature robot fitted with a drill guide that provides rigid mechanical guidance for hand-held drilling of the distal screws' pilot holes. The robot is automatically positioned so that the drill guide and nail distal locking axes coincide, using a single fluoroscopic X-ray image. Since the robot is rigidly attached to the intramedullary nail or bone, no leg immobilization or real-time tracking is required. We describe the system and protocol and present a method for accurate and robust drill guide and nail hole localization and registration. The in vitro system accuracy experiments for fronto-parallel viewing show a mean angular error of 1.3 degrees (std = 0.4 degrees ) between the computed drill guide axes and the actual locking holes axes, and a mean 3.0 mm error (std = 1.1 mm) in the entry and exit drill point, which is adequate for successfully locking the nail. PMID- 15889551 TI - Direct reconstruction of kinetic parameter images from dynamic PET data. AB - Our goal in this paper is the estimation of kinetic model parameters for each voxel corresponding to a dense three-dimensional (3-D) positron emission tomography (PET) image. Typically, the activity images are first reconstructed from PET sinogram frames at each measurement time, and then the kinetic parameters are estimated by fitting a model to the reconstructed time-activity response of each voxel. However, this "indirect" approach to kinetic parameter estimation tends to reduce signal-to-noise ratio (SNR) because of the requirement that the sinogram data be divided into individual time frames. In 1985, Carson and Lange proposed, but did not implement, a method based on the expectation maximization (EM) algorithm for direct parametric reconstruction. The approach is "direct" because it estimates the optimal kinetic parameters directly from the sinogram data, without an intermediate reconstruction step. However, direct voxel wise parametric reconstruction remained a challenge due to the unsolved complexities of inversion and spatial regularization. In this paper, we demonstrate and evaluate a new and efficient method for direct voxel-wise reconstruction of kinetic parameter images using all frames of the PET data. The direct parametric image reconstruction is formulated in a Bayesian framework, and uses the parametric iterative coordinate descent (PICD) algorithm to solve the resulting optimization problem. The PICD algorithm is computationally efficient and is implemented with spatial regularization in the domain of the physiologically relevant parameters. Our experimental simulations of a rat head imaged in a working small animal scanner indicate that direct parametric reconstruction can substantially reduce root-mean-squared error (RMSE) in the estimation of kinetic parameters, as compared to indirect methods, without appreciably increasing computation. PMID- 15889552 TI - Estimation of extraction fraction (EF) and glomerular filtration rate (GFR) using MRI: considerations derived from a new Gd-chelate biodistribution model simulation. AB - Previous reports have described the use of magnetic resonance imaging (MRI) to estimate single-kidney extraction fraction (EF) and glomerular filtration rate (GFR), by measuring the concentration difference of intravenously injected Gd chelate ([Gd]) in the renal artery and renal vein from measurements of blood T1. Problematic is the fact that [Gd] measurements in the renal artery are often inaccurate due to the small size, tortuousness and motion of the vessel. Consequently, the [Gd] in the inferior vena cava (IVC) below the renal vein ostia (i.e., the infrarenal IVC) has been used instead of the renal artery [Gd], based on the assumption that the [Gd] in the infrarenal IVC is the same as it is in the renal artery. However, this assumption has neither been theoretically nor experimentally investigated. Herein, we describe new difference and differential equation pharmacological models that can predict the biodistribution of Gd chelate throughout the extracellular space. Assuming known average normal blood flows and GFR, our models predict that the infrarenal IVC [Gd] is 3.2% to 4.7% greater than the renal artery [Gd], and that the EF estimate using this IVC measurement is overestimated by 14.2%-20.0%. To support these predictions, algebraic equations are derived which show that the infrarenal IVC must develop a relatively high [Gd] in order to satisfy Gd flux constraints within the vascular system. These results suggest that the infrarenal IVC [Gd] is not a valid substitute for the renal artery [Gd]. PMID- 15889553 TI - Comparison between MAP and postprocessed ML for image reconstruction in emission tomography when anatomical knowledge is available. AB - Previously, the noise characteristics obtained with penalized-likelihood reconstruction [or maximum a posteriori (MAP)] have been compared to those obtained with postsmoothed maximum-likelihood (ML) reconstruction, for emission tomography applications requiring uniform resolution. It was found that penalized likelihood reconstruction was not superior to postsmoothed ML. In this paper, a similar comparison is made, but now for applications where the noise suppression is tuned with anatomical information. It is assumed that limited but exact anatomical information is available. Two methods were compared. In the first method, the anatomical information is incorporated in the prior of a MAP algorithm and is, therefore, imposed during MAP-reconstruction. The second method starts from an unconstrained ML-reconstruction, and imposes the anatomical information in a postprocessing step. The theoretical analysis was verified with simulations: small lesions were inserted in two different objects, and noisy PET data were produced and reconstructed with both methods. The resulting images were analyzed with bias-noise curves, and by computing the detection performance of the nonprewhitening observer and a channelized Hotelling observer. Our analysis and simulations indicate that the postprocessing method is inferior, unless the noise correlations between neighboring pixels are taken into account. This can be done by applying a so-called prewhitening filter. However, because the prewhitening filter is shift variant and object dependent, it seems that MAP reconstruction is the more efficient method. PMID- 15889554 TI - Submillimeter measurement of cup migration in clinical standard radiographs. AB - Assessing the displacement of bony implants is an important topic in arthroplasty, particularly in total hip replacement (THR). The observation of the migration is supposed to provide an insight into the fixation of the implant. Diagnostic standard radiographs of the pelvis are an advantageous data source for this purpose. The previous methods based on these images, however, lack of a thorough consideration of their projective nature. They do, hence, not reach the desired precision, which should lie in the submillimeter range to allow a detection of migration in the first one or two years after implantation. The aim of the work presented here was, therefore, a method for measuring the distance of the artificial hip socket to the bone with an error of less than 0.5 mm. The approach has been on the one hand to define the bone-cup distance measured in the radiograph so that the variability of the intrinsic and extrinsic parameters at exposure has a minimal impact. On the other, specialized matching techniques are applied in order to optimize the localization of the necessary bony landmarks and the cup in the X-ray image. The coordinates of the bony landmarks are determined by means of a template matching algorithm. The position of the implant is estimated by intensity-based registration using the cup's CAD-model. The method was validated theoretically, experimentally, and clinically. In the clinical radiographs, the standard deviation of the migration measurements resulted to be 0.28 mm when using only natural bony landmarks. The implantation of a bony marker was found to increase the precision to a standard deviation of 0.20 mm. The interobserver variability in the two cases was estimated to be 0.11 mm and 0.04 mm. PMID- 15889555 TI - Distributed vector processing of a new local multiscale Fourier transform for medical imaging applications. AB - The recently developed S-transform (ST) combines features of the Fourier and Wavelet transforms; it reveals frequency variation over both space and time. It is a potentially powerful tool that can be applied to medical image processing including texture analysis and noise filtering. However, calculation of the ST is computationally intensive, making conventional implementations too slow for many medical applications. This problem was addressed by combining parallel and vector computations to provide a 25-fold reduction in computation time. This approach could help accelerate many medical image processing algorithms. PMID- 15889556 TI - Orthopaedic infections. PMID- 15889557 TI - Orthopaedic infections: organisms and antibiotic sensitivity. AB - Staphylococcus aureus infection remains the commonest organism causing musculoskeletal infection and antibiotic is the mainstay of treatment apart from adequate and appropriate surgical intervention. The exact figure of antibiotic resistance in orthopaedic practice is not known but it is expected to be higher than previously reported as the use of antibiotics is rampant. Its sensitivity to various antibiotics differs from one center to another making local surveillance necessary. From 66 patients with musculoskeletal infections studied in our centre, Staphylococcus aureus was cultured in 50-65% of patients, depending on the sample taken. Fifteen percent of this were methicillin resistant Staphylococcus aureus (MRSA). Staphylococcus aureus was found to be sensitive to cloxacillin in 95% of patients' sample. MRSA remained highly sensitive to vancomycin, clindamycin and fucidic acid. PMID- 15889558 TI - Comparison of nerve graft and artificial conduits for bridging nerve defects. AB - A study of nerve regeneration through a 1cm defect in the peroneal component of the sciatic nerve was performed on sixteen rabbits. Either silicone or polytetrafluoroethylene (PTFE) tubes or nerve graft were used to bridge the defect and the opposite limb was not operated upon. The rabbits that underwent nerve grafting had favourable findings. In the PTFE group, a nerve-like structure was seen at the former gap site and histology confirmed viable axons within the tubes and distal to the repair site. In the silicone tube group, there were no myelinated axons demonstrated. The axonal count for the grafted nerves and the nerves repaired with PTFE tube are on average 80.4% and 38.2% of that of the unoperated nerve, respectively. On average, the percentage anterior compartment muscle weight (expressed as a percentage of the unoperated limb) for the silicone, PTFE and nerve graft groups are 42.3%, 42.1%, and 72.7% respectively. The results show that although, PTFE conduits can bridge a nerve defect of 1cm, nerve grafting provides a superior and more predictable outcome. PMID- 15889559 TI - Oropharyngeal flora changes in patients with head and neck malignancy post radiotherapy. AB - Radiotherapy has been recognized as a valuable modality of treatment in the management of head and neck cancers. It can have a direct bactericidal effect on the normal flora of the oropharynx. The objective of this study is to determine the changes in the oropharyngeal flora after external beam radiation. This prospective non randomized control study was performed to aid in identification of organisms involved in sepsis, as well as aid in choosing appropriate antibiotics for surgical procedures in irradiated patient. Forty patients with various head and neck malignancy and thirty control patients were selected. Oropharyngeal swabs were taken prior to radiotherapy, at the end and one month after radiotherapy. A single swab was taken from the control group. A full bacteriological analysis was performed. There was a statistically significant decrease in Alpha Hemolytic Streptococci and Neisseria species post radiotherapy. B Proteus and Candida Albicans showed a statistical significant increase in patients with head and neck cancer post radiotherapy. These changes remained even one month after radiotherapy. PMID- 15889560 TI - Percutaneous dilational tracheostomy--a 3 year experience in a general hospital in Malaysia. AB - All percutaneous tracheostomies performed in the general intensive care unit (ICU), Hospital Sultanah Aminah, Johor Bahru, Malaysia, from July 1999 to June 2002 were studied. The tracheostomies were performed as an elective bedside technique in the ICU. A total of 352 percutaneous tracheostomies were performed. Eighty-eight percent of the tracheostomies were completed within 15 minutes. The most common complication was bleeding which occurred in 52 patients (14.7%). The rest of the complications encountered were:- transient hypoxia twelve (3.4%), inability/ difficulty to insert tracheostomy tube eight (2.3%), false passage four (1.1%), transient hypotension two (0.6%), pneumothorax two (0.6%), peristomal infection two (0.6%), subcutaneous emphysema one (0.3%), cuff rupture one (0.3%), oesophageal cannulation one (0.3%), and granuloma formation one (0.3%). Conversion to conventional tracheostomies were performed on 7 patients (2%). There was one unfortunate death related to percutaneous tracheostomy. In conclusion, percutaneous dilational tracheostomy can be used safely to manage the airway of critically ill patients. PMID- 15889561 TI - The effect of a user-guided request form for chest radiographs in a neonatal intensive care unit. AB - The objectives of this prospective, observational study were to determine the current indications of requesting chest radiograph in sick infants in a neonatal intensive care unit (NICU) and the effect of a user-guided request form for chest radiographs of sick infants. During the three-month study period, a total of 423 chest radiographs were requested on 159 sick newborn infants in the intensive area of this NICU. A majority (55.6%) of these chest radiographs were performed to verify positions tips of either central catheters (27.4%) or endotracheal tubes (28.1%). The number of chest radiographs done during the period when the user-guided request forms were utilised was significantly lower (1.24 per patient) than before (1.37 per patient) or after (1.58 per patients) the period when these forms were in use (p=0.01). The rate of radiological abnormalities detected in radiographs requested to verify position of tips of endotracheal tubes were significantly greater during the period when user-guided forms were used than when they were not (p=0.01). A significantly higher proportion of changes in management were instituted when the user-guided forms were in use than during the period when they were not used (p=0.03). We conclude that a user guided radiographic request form helps doctors in NICU to carry out their management more effectively. PMID- 15889562 TI - The correlation between cytopenia and esophageal varices in patients with liver cirrhosis. AB - This retrospective study analysed the case records of 200 patients in the Department of Gastroenterology, Singapore General Hospital from February 2000 to January 2001 who had liver cirrhosis and underwent gastroscopy for the detection of varices. The aim of this study was to determine any relationship between leucopenia, thrombocytopenia and the occurrence of esophageal varices in a cirrhotic population. Our results showed that the diagnostic yield of varices grade 2 and 3 was 6.3% if platelet count was > 150,000/mm3, 25% if platelet count was 100,000 to 150,000/mm3, 38.9% if platelet count was 50,000-99,000/mm3 and 100% if platelet count was <50,000/mm3. Similarly, the diagnostic yield of varices grade 2 and 3 was 19.4% if total white count was > 4,000/mm3, 66.7% if total white count was 3,000- 4,000/mm3 and 94.8% if total white count was < 3,000/mm5. We conclude that thrombocytopenia and leucopenia can be used to stratify risk for occurrence of esophageal varices in cirrhotic patients and gastroscopy will have a high yield for varices when platelet count is < 150,000/mm3 or total white is < 4,000/mm. PMID- 15889563 TI - Audits in general practice by medical students. AB - This paper reports the implementation, findings and feedback of the audit project of the general practice (GP) module carried out in the fifth year of the MBChB (Sheffield) twinning programme with the Perak College of Medicine. After training, each student with his/her GP tutor planned and conducted the audit. All 28 students (year 2002) satisfactorily completed their audit projects. Fifty percent did an audit of hypertension, 36% on diabetes, 7% on asthma and one each (3.5%) on upper respiratory tract infection and client satisfaction. It was the GP tutors first experience at audit in their clinics. The majority of indicators of care audited did not meet the set target standards. PMID- 15889564 TI - Clinical and endoscopic features of peptic ulcer bleeding in Malaysia. AB - The characteristics of patients and the endoscopic features of 196 patients with bleeding peptic ulcer in a multi-ethnic population were investigated. There was a male preponderance (M: F= 6.3: 1) and their mean age was 63.5 years. The prevalence of peptic ulcer bleeding in the Malays and Indians was similar to the ethnic distribution of population. However, the Chinese were over represented. Nearly 40% of patients studied had at least one co-existing medical illness. Hypertension and ischaemic heart disease were the most common diseases. History of non-steroidal anti-inflammatory drug usage was identified in 48% of the patients and it was the commonest risk factor associated with bleeding ulcers. More than 80% of bleeding ulcers were located in the duodenum and the pylorus. Endoscopic features of active bleeding or recent bleed were identified in more than 60% of the patients. The study notes that bleeding peptic ulcer is a serious and a potentially life threatening condition. It is a disease of the elderly and, with the steadily increasing elderly population in the country, the admissions rates of peptic ulcer bleeding is expected to rise. There is a need to plan for appropriate technical support, critical care facilities and expertise to avoid unacceptable outcomes. PMID- 15889565 TI - Vitamin E deficiency reduced lumbar bone calcium content in female rats. AB - Vitamin E deficiency has been found to impair bone calcification. This study was done to determine the effects of vitamin E deficiency and supplementation on parathyroid hormone, i.e. the hormone involved in bone regulation. Female Sprague Dawley rats were divided into 4 groups: 1) normal rat chow (RC), 2) vitamin E deficiency (VED), vitamin E deficient rats supplemented with 3) 60 mg/kg alpha tocotrienol (ATT) and 4) 60 mg/kg (alpha-tocopherol (ATF). Treatment was carried out for 3 months. Vitamin E deficiency caused hypocalcaemia during the first month of the treatment period, increased the parathyroid hormone level in the second month and decreased the bone calcium content in the 4th lumbar bone at the end of the treatment. Vitamin E supplementation (ATT and ATF) failed to improve these conditions. The bone formation marker, osteocalcin, and the bone resorption marker, deoxypyridinoline did not change throughout the study period. In conclusion vitamin E deficiency impaired bone calcium homeostasis with subsequent secondary hyperparathyroidism and vertebral bone loss. Replacing the vitamin E with pure ATF or pure ATT alone failed to correct the changes seen. PMID- 15889566 TI - The prognostic value of C-reactive protein (CRP) levels in patients with acute ischaemic stroke. AB - Increasing evidence suggests that inflammation plays an important role in the development of both cardiovascular and cerebrovascular events. Recently C reactive protein (CRP) levels have been reported to be a prognostic factor for cerebrovascular and cardiovascular events. The main objective of the study was to evaluate the prognostic value of CRP levels in a first ever ischaemic stroke at one month. All ischaemic stroke patients who were admitted to Hospital Universiti Kebangsaan Malaysia (HUKM) between May 2002 and July 2002 were eligible for the study. CRP levels were taken within 72 hours after an acute ischaemic stroke. The functional ability was assessed using the Barthel Index (BI) after one month of stroke. During the study period 84 patients were admitted to HUKM with the diagnosis of ischaemic stroke; 49 patients were enrolled and 35 were excluded. Twenty-nine patients (59.2%) had elevated CRP levels (median 1.64+/-3.07 mg/dL, range 0.06 to 16.21 mg/dL). Elevated CRP levels were found to be a predictor of severe functional disability (BI<5) and were also associated with larger infarcts. In conclusion, elevated CRP levels are associated with poorer functional outcome and predict a larger infarct size. PMID- 15889567 TI - Lupus band test in systemic lupus erythematosus. AB - The usefulness of the direct immunofluorescent antibody technique--lupus band test--for the diagnosis of systemic lupus erythematosus (SLE) has been well established. The aims of the study were to determine the prevalence of the LBT at various sites of the skin in a cross section of patients with SLE and its correlation with disease activity. The LBT was demonstrated in 64% of skin lesions, 63% in non-lesional sun-exposed (NLSE) skin and 25% in non-lesional sun protected (NLSP) skin. The prevalence of the LBT in lesional and NLSE groups was significantly different from the NLSP group (p = 0.03 and 0.005 respectively). There was a significant correlation between the presence of a positive LBT in NLSE skin with the presence of the LE cell phenomenon (p = 0.04) and anti - ds DNA antibody (0.02). In addition, there was a significant correlation between IgG LBT in the NLSE skin with serum hypocomplementaemia (p = 0.03) and anti - ds DNA antibody (p = 0.04). Other than these, no significant correlation was detected between the LBT from the 3 sites with overall clinical activity, renal disease, active skin lesions, or other laboratory indices of activity. These findings suggest that the LBT is mainly indicated as a diagnostic tool and has little role in assessing disease activity. PMID- 15889568 TI - Medication compliance among renal transplant patients: a Hospital Kuala Lumpur experience. AB - This survey aims to identify prevalence, reasons and predictors of noncompliance among renal transplant patients followed up in Hospital Kuala Lumpur (HKL). All adult renal transplant patients who were at least 6 months post transplant were recruited from 10/2001 till 5/2002. Patients who consented were interviewed by a medical doctor or research assistant based on questionnaire. Noncompliers were defined as those who missed or self adjusted any dose of immunosuppressant within the preceding 4 weeks. Inter-rater agreement was assessed prior by Kappa (K) scores and they were acceptable. Out of 304 patients, 246 patients volunteered; of whom 144 (58.5%) were males. Twenty-one (9.3%) were noncompliers. Reasons for noncompliance included forgetfulness (n=8), financial constraints (n=1), fear of rejection (n=1), side effects (n=9), decision not to take (n=6), difficulty in breaking medication into correct dosages (n=1). Significant predictors of noncompliance were longer duration of transplant noncompliance to other drugs, regular use of nonprescription drugs; the lack of symptoms of fat facial cheeks and infection. Surveillance for noncompliance should not be relaxed as its predictors are diverse and persistent, especially in those who are at high risks. PMID- 15889569 TI - Change in infection control practices and awareness of hospital medical staff in the aftermath of SARS. AB - Severe Acute Respiratory Syndrome (SARS) epidemic illustrated the crucial role of infection surveillance and control measures in the combat of any highly transmissible disease. We conducted an interview survey of 121 medical staff 145 doctors, 46 staff nurses and 30 medical assistants) in a state hospital in Malaysia three months after the end of SARS epidemic (from October to December 2003). Staff was grouped according to those directly involved in the care of suspected SARS patients [S+ group n=41] and those who were not [S- group; n=80]. On hand washing following sneezing, coughing and touching patients, the proportions of medical staff that reported an increase after the SARS crisis were 22.3%, 16.5% and 45.5% respectively. On wearing masks, gloves, and aprons when meeting potentially infectious patients, the proportions that reported an increase were 39.7%, 47.1% and 32.2% respectively. Significantly more staff in S+ than S- group reported these increases. Sixty percent of staff was aware of changes in hospital infection control policies after SARS; 93.4% was aware of notifying procedures, and 81.8% was aware of whom to notify in the hospital. Regarding infection isolation ward, Infectious Control Nurse and Infection Control Committee Chairman in the hospital, the proportions of staff that could correctly name them were 39.7%, 38.3% and 15.7% respectively. Significantly more in S+ than S- group could do so. However, more than half the staff claimed ignorance on the knowledge of infection isolation ward (56.2%), Infection Control Nurse (57.9%) and Chairman (65.3%). Our findings demonstrated that SARS crisis had some positive impact on the infection control practices and awareness of medical staff especially on those with direct SARS involvement. Implications for future control of infectious diseases are obvious. PMID- 15889570 TI - Outcome of infection following internal fixation of closed fractures. AB - A retrospective study was done in 30 patients with infected closed fractures treated initially by open reduction and internal fixation. Nineteen fractures involved the femur, 8 the tibia, 2 the radius and 1 the ulna. Twenty-five were diaphyseal and 5 metaphyseal. Twenty-two fractures had initially been treated with plating and 8 with intramedullary nailing. Thirteen patients presented in the acute phase and 17 in the subacute phase of infection. Twenty-two patients presented with low grade infection and 8 with florid infection. The commonest organism isolated was Staphylococcus aureus (80%), of which fifty four percent was resistant to Methicillin (MRSA). This study showed that 77% of infected fractures with a stable implant united even in the presence of infection. PMID- 15889571 TI - Hepatitis A outbreak in Hulu Langat District, Selangor State, Malaysia during April--October 2002. AB - A Hepatitis A outbreak occurred in Mukim Hulu Langat, Hulu Langat district from April 2002 to October 2002. Of the 51 cases notified, most were among students and the ethnic groups involved were Malays and the Orang Asli (local indigenous community). Epidemiological investigations revealed that the cases were localized along rivers used for recreational activities in this area. River water analysis indicated human faecal contamination and it was believed that the contamination was due to the Orang Asli community living upstream. This occurred due to lack of toilet facilities, water at point of use and the existing traditional practices of the Orang Asli community. Control measures instituted were intense health education to the Orang Asli to avoid using the rivers for defecation, multi agency efforts to provide sanitary toilets and adequate water to the villages affected. Future measures include conducting a sero- prevalence survey to determine the feasibility of Hepatitis A immunization to the susceptible population in this area. The outbreak that began in April 2002 was controlled by October 2002. PMID- 15889572 TI - Extra-corporeal shock-wave therapy in the treatment of non-unions. AB - It has been at least a decade since the introduction of extra-corporeal shock wave treatment (ESWT) for the treatment of non-unions. Despite conflicting opinions in the literature, it is recently experiencing a revival. This paper reports our experience with Ossa Tron, which produces shock-waves electro hydraulically, on 5 patients. The two successful cases experienced remarkable results of union at an average of 22 weeks after ESWT. The remaining three had disappointing results. A description of the study and a note on the essential issues are presented. PMID- 15889573 TI - Benign teratoma of the nasal cavity. AB - A teratoma is a true neoplasm composed of an assortment of tissue often alien to the site from it arises. Teratomas of the head and neck are exceedingly rare and usually present in the neonatal period. Most of these are found in the cervical region and nasopharynx. Though most tumors are benign, they are often malignant with regards to the location. Here we present a rare case of benign teratoma arising from the nasal septum and presenting for the first time in a young adult. Appropriate literature is reviewed. PMID- 15889574 TI - Actinomyces infection of the mastoid: a rare entity. AB - A rare case of mastoid infection caused by actinomyces israelii is presented. This patient underwent exploratory mastoidectomy followed by long term oral pencillin. She responded well to the treatment and has been asymptomatic on follow up to date. PMID- 15889575 TI - Multifocal choroiditis: ocular TB or side-effects of anti-TB therapy? AB - A patient with end-stage renal failure secondary to polycystic kidney disease was seen in the Eye Clinic for a corneal abrasion. Incidental fundal examination revealed bilateral multiple, small, raised, pale yellow sub-retinal nodules. Past medical history of inadequately treated pulmonary tuberculosis was obtained. Following initiation of anti-tuberculous therapy, the choroidal lesions resolved. PMID- 15889576 TI - Right internal carotid pseudoaneurysm mimicking a retropharyngeal abscess in a child. AB - A 7-year-old girl with tonsillar infection with antibiotics. Two weeks later, there was a right sided neck lump. Computed tomography scans demonstrated a predominantly hypodense right retropharyngeal area with peripheral enhancement and mass effect. There was intense enhancement within the postero-superior aspect of the lesion which was continuous with the right internal carotid artery. Ultrasound demonstrated tapering of the right internal carotid artery. Magnetic resonance imaging and magnetic resonance arteriography showed a right internal carotid artery pseudoaneurysm. Surgical exploration confirmed the finding. This case highlights an unusual presentation of an internal carotid pseudoaneurysm and how imaging provided the diagnosis. PMID- 15889577 TI - Nocardia infection of the mastoid in an immunocompromised patient. AB - A rare case of Nocardia infection of mastoid is presented in an immunocompromised patient. PMID- 15889578 TI - Atypical antipsychotic induced obsessive-compulsive symptoms. PMID- 15889579 TI - Wrong conclusion when the statistical test is incorrect. PMID- 15889580 TI - Gestational trophoblastic disease. AB - Gestational trophoblastic disease is a spectrum of pregnancy disorder arising from the placental trophoblastic tissues. It characterised by the secretion of a distinct tumour marker, the beta-HCG. This condition is highly curable even in the presence of metastasis. The incidence of this disease is higher in the Asian population. The major well-established risk factors for gestational trophoblastic disease are advanced maternal age and a past history of gestational trophoblastic disease. Common clinical presentations include vaginal bleeding in early trimester, uterus larger than gestational age, absence of fetal parts after 20 weeks of gestation. Ultrasonography is a reliable non-invasive tool for diagnosis of gestational trophoblastic disease in the clinical setting. All placental tissue following miscarriage or curettage should have histopathological evaluation to exclude gestational trophoblastic disease. Since this group of disorders is one of the highly curable neoplasms, early diagnosis and prompt treatment is necessary. PMID- 15889581 TI - Increasing individual upper alpha power by neurofeedback improves cognitive performance in human subjects. AB - The hypothesis was tested of whether neurofeedback training (NFT)--applied in order to increase upper alpha but decrease theta power--is capable of increasing cognitive performance. A mental rotation task was performed before and after upper alpha and theta NFT. Only those subjects who were able to increase their upper alpha power (responders) performed better on mental rotations after NFT. Training success (extent of NFT-induced increase in upper alpha power) was positively correlated with the improvement in cognitive performance. Furthermore, the EEG of NFT responders showed a significant increase in reference upper alpha power (i.e. in a time interval preceding mental rotation). This is in line with studies showing that increased upper alpha power in a prestimulus (reference) interval is related to good cognitive performance. PMID- 15889582 TI - Autonomic nervous system activity during actual and mentally simulated preparation for movement. AB - The aim of this study was to compare actual versus mentally simulated preparation for a complex motor skill. Two behavioral periods are observed during weightlifting: (i) an initial phase in which the subject standing behind the bar is thought to focus his attention on forthcoming execution and (ii) a second phase between hands/bar contact and execution during which the subject is thought to increase activation. Such mental processes accompanying behavioral sequences are correlated with autonomic nervous system activity, phasic responses corresponding to allocation of attentional resources, and tonic variations related to increasing general activation. To study mental processes during preparation for action, 12 subjects performed actual and imagined preparation phases of execution. Six autonomic variables were measured continuously. Skin potential (chi2 = 0.16), skin temperature amplitude (Z = -0.66) and duration (Z = -1.78), skin blood flow amplitude (Z = -0.56) and duration (Z = -1.51), respiratory frequency amplitude (Z = -0.14) and duration (Z = -0.13), and duration of heart rate response (Z = -1.25) were shown to be comparable (p > .05), whatever the modality of preparation. However, during mentally simulated preparation, skin resistance response was shorter than in actual preparation (Z = -2.12, p < .05), thus attesting to a weaker load, whereas lower decrease in heart rate was elicited (Z = -1.96, p < .05). This may be explained by this particular experimental condition because mental preparation would not lead to actual action. Such autonomic variables could be used as feedback to improve performance. PMID- 15889583 TI - A behavioral group treatment program for menopausal hot flashes: results of a pilot study. AB - In the present study, we tested the effectiveness of a cognitive-behavioral group treatment (CBGT) for hot flashes in menopausal women. Treatment was administered over 8, 90 min weekly sessions and consisted of education, relaxation training and cognitive restructuring. Nineteen women meeting STRAW staging criteria for the menopause transition (stages -1 to +1) were randomly assigned to immediate or delayed treatment (wait list) and were asked to monitor their hot flashes and night sweats prospectively. They also completed questionnaires, including the Women's Health Questionnaire and the Menopause Specific Quality of Life Questionnaire to determine psychosocial benefits of treatment. Results suggested that the CBGT was moderately successful in reducing the frequency of total vasomotor symptoms [F (1, 17) = 6.16, p < .01], as measured by daily symptom diaries. While there were arithmetic improvements in psychosocial functioning in this sample, these results were not significant. Despite the limitations of small sample size and possible placebo effect, this pilot study supports the notion that cognitive-behavioral interventions aimed at reducing vasomotor symptoms may be of value for menopausal hot flashes when administered in a small-group format. PMID- 15889584 TI - Calibration of ADHD assessments across studies: a meta-analysis tool. AB - When analyzed separately, data from small studies provide only limited information with limited clinical generalizability, due to small sample size, differing assessments, and limited scope. In this methodological paper we outline a theoretical framework for performing meta-analysis of data obtained from disparate studies using disparate tests, based on calibration of the data from such studies and tests into a unified probability scale. We apply this method to combine the data from five studies examining the diagnostic abilities of different assessments of Attention Deficit/Hyperactivity Disorder (ADHD), including behavioral rating scales and EEG assessments. The studies enrolled a total of 111 subjects, 56 ADHD and 55 controls. Each individual study had a small sample focused on a specific age/gender group, for example 8 boys ages 6-10, and generally had insufficient power to detect statistically significant differences. No gender, or age comparisons were possible within any single study. However, when calibrated and combined, the data resulted in a clear separation between ADHD versus non-ADHD groups in males below the age of 16 (p < 0.001), males above the age of 16, (p = 0.015), females below the age of 16, (p = 0.0014), and females above the age of 16, (p = 0.0022). We conclude that if data from various studies using various tests are made comparable, the resulting combined sample size and the increased diversity of the combined sample lead to increased significance of the statistical tests and allow for cross-sectional comparisons, which are not possible within each individual study. PMID- 15889585 TI - The effect of the definition of menstrually-related headache on the response to biofeedback treatment. AB - Twelve women with either pure migraine headache (HA) or a combination of migraine and tension-type HA monitored HA activity over two menstrual cycles while noting onset of menstruation and onset of ovulation. They then received 12 sessions of thermal biofeedback (TBF) with adjunctive autogenic training. Six women with only tension-type HA participated in similar monitoring before receiving 9 sessions of progressive relaxation training. Results for those with vascular HA showed a significant reduction in HA activity and a reduction in medication taken for HAs. Those with tension-type HA did not respond significantly to the relaxation training. Depending upon how one defined menstrually-related HAs among those with vascular HA, there either was, or was not, a differential effect of TBF on menstrual-cycle-related HA. PMID- 15889586 TI - Biofeedback and dance performance: a preliminary investigation. AB - Alpha-theta neurofeedback has been shown to produce professionally significant performance improvements in music students. The present study aimed to extend this work to a different performing art and compare alpha-theta neurofeedback with another form of biofeedback: heart rate variability (HRV) biofeedback. Twenty-four ballroom and Latin dancers were randomly allocated to three groups, one receiving neurofeedback, one HRV biofeedback and one no intervention. Dance was assessed before and after training. Performance improvements were found in the biofeedback groups but not in the control group. Neurofeedback and HRV biofeedback benefited performance in different ways. A replication with larger sample sizes is required. PMID- 15889587 TI - Hot flash, hot topic: conceptualizing menopausal symptoms from a cognitive behavioral perspective. AB - While most healthy women report that the menopausal transition is nondistressing, a subset of women does report that symptoms significantly interfere in their lives. The most common reason that women seek treatment during this time is for vasomotor symptoms, namely, hot flashes and night sweats. Research has suggested that reports of distress during flashing are only weakly related to more objective measures of the flash, including duration and frequency and that differences in treatment-seeking during the menopausal transition may be better accounted for by differences in symptom awareness mediated by a variety of personality and stress factors. This paper discusses hot flashes and night sweats from a cognitive-behavioral perspective, taking into account individual difference variables that may also affect the experience of menopausal symptoms. PMID- 15889588 TI - Biofeedback in the treatment of phantom limb pain: a time-series analysis. AB - Phantom limb pain (PLP) is a noxious, painful sensation that is perceived to occur in an amputated limb. It has been reported to occur in up to 85% of amputees. This pilot study examined the effectiveness of biofeedback in the treatment of nine individuals with PLP who received up to seven thermal/autogenic biofeedback sessions over the course of 4-6 weeks. Pain was assessed daily using the visual analog scale (VAS), the sum of the sensory descriptors, and the sum of the affective descriptors of the McGill short form. Interrupted time-series analytical models were created for each of the participants, allowing biofeedback sessions to be modeled as discrete interventions. Analyses of the VAS revealed that a 20% pain reduction was seen in five of the nine patients in the weeks after session 4, and that at least a 30% pain reduction (range: 25-66%) was seen in six of the seven patients in the weeks following session 6. Sensory descriptors of pain decreased more than the affective pain descriptors. These preliminary results provide some support for the use of biofeedback in the treatment of PLP and indicate the need for further, definitive study. PMID- 15889589 TI - [Effects of circumferential rigid wrist orthoses in rehabilitation of patients with radius fracture at typical site]. AB - BACKGROUND: The use of orthoses is a questionable rehabilitation method for patients with the distal radius fracture at typical site. The aim of this study was to compare the effects of the rehabilitation on patients with radius fracture at the typical site, who wore circumferential static wrist orthoses, with those who did not wear them. METHODS: Thirty patients were divided into 3 equal groups, 2 experimental groups, and 1 control group. The patients in the experimental groups were given the rehabilitation program of wearing serially manufactured (off-the-shelf), as well as custom-fit orthoses. Those in the control group did not wear wrist orthoses. Evaluation parameters were pain, edema, the range of the wrist motion, the quality of cylindrical, spherical, and pinch-spherical grasp, the strength of pinch and hand grasp, and patient's assessment of the effects of rehabilitation. RESULTS: No significant difference in the effects of rehabilitation on the patients in experimental groups as opposed to control group was found. Patients in the first experimental group, and in control group were more satisfied with the effects of rehabilitation, as opposed to the patients in the second experimental group (p<0.05). CONCLUSION: The effects of circumferential static wrist orthoses in the rehabilitation of patients with distal radius fracture at the typical site were not clinically significant. There was no significant difference between the custom and off-the-shelf orthoses. PMID- 15889590 TI - [Risk factors for the development of hospital infections in the intensive care units]. AB - BACKGROUND: Patients admitted to intensive-care units (ICU) are at a high risk of nosocomial infections (NI) due to susceptibility associated with severity of their condition, but also the invasive medical procedures they undergo. AIM: To determine the frequency of NI at the ICU of the General Hospital Uzice, and to identify the risk factors for their development. METHODS: A prospective surveillance study of NI, conducted between June 27. and December 31 2001, included 914 patients who spent at least 24 hours in the ICU (total of 2 615 days). The surveillance of NI in the ICU was carried out daily. Follow-up period covered the time from the ICU admission to 48 hours after the ICU discharge. To assess risk factors for NI, we performed a case-control study. The variables measuring of extrinsic and intrinsic risk factors for NI were collected. RESULTS: In a six-month prospective surveillance study, the incidence of NI was 16.7% or 58.5 per 1,000 patient-day, respectively. The most frequent were the infections of the surgery wounds (32.6%), urinary tract infections (23.5%), and infections of the blood (7.1%). The identified independent risk factors for NI were: surgical intervention (OR = 5.74; CI = 2.01-16.41), endotracheal tubes (OR = 3.40; CI = 1.07-10.89), cystoscopy (OR = 2.35; CI = 1.38-4.02), obesity (OR = 1.98; CI = 1.27-3.11), and the duration of the infusion (OR = 1.34; CI = 1.23 1.46). CONCLUSIONS: The most important risk factors for NI at ICU were surgical interventions and endotracheal tubes. PMID- 15889591 TI - Very early increase in nitric oxide formation and oxidative cell damage associated with the reduction of tissue oxygenation is a trait of blast casualties. AB - BACKGROUND: In our previous experimental studies, we found evidence for the increase of nitric oxide (NO) formation immediately after blast injury. In the present study we investigated whether NO overproduction was a trait for the period immediately after blast injury in humans. Concomitant metabolic disturbances were also studied, and compared to the alterations in other traumatized patients. METHODS: Blast casualties (group B, n = 13), surgical patients with the hip replacement or fractures, not exposed to blast effects (group S, n = 7) and healthy volunteers as controls (group C, n = 10), were examined. Both arterial and venous blood samples were taken within 6 hours, and 24 hours after blast injuries or surgical procedures, respectively. Plasma levels of nitrite/nitrate (NOx), superoxyde anion (O2-), sulfhydrils (SH), malondialdehyde (MDA) as well as acid-base status and other biochemical parameters (glucose, urea, creatinine, total proteins, albumin) were measured. RESULTS: Significant, but transient increase in plasma NOx levels occurred only in group B. It was associated with the significant increase of hemoglobin oxygen (sO2) saturation of the venous blood and the concomitant decrease of its arterial -venous difference. In group S the venous sO2 decreased, its arterial--venous difference increased, while NOx levels were within the control limits. In both groups, other parameters of arterial acid-base status were kept within the control limits throughout the examined period. The decrease of SH levels were similar in the examined groups, while the increase of O2- was greater in group B. CONCLUSION: Early NO overproduction was a trait of blast injuries in humans, contributing to the reduction of tissue the oxygenation and intensifying the oxidative cell damage that had to be considered in the therapy of casualties with blast injuries. These alterations were different from those observed in other surgical patients without blast injuries. PMID- 15889592 TI - Quantitative analysis of the nephron during human fetal kidney development. AB - BACKGROUND: The development of human kidney is a complex process. The number, shape, size, and distribution of nephrons as functional units in a kidney, provide some important information about the organization of the kidney. The aim of this study was to extend the knowledge of the developing human kidney by studying nephrons in the kidney's cortex during gestation. METHODS: Kidney tissue specimens of 32 human fetuses, the gestational age from IV lunar month (LM IV) to LM X, were analysed. Specimens were divided in ten groups based on gestational age. Stereological methods were used at the light microscopic level to estimate the volume densities of the corpuscular and tubular components of the nephron in the cortex of the developing human kidney. RESULTS: Nephron polymorphism was the main characteristic of the human fetal kidney during development. In younger fetuses, just below the renal capsule, there was a wide nephrogenic zone. It contained the condensed mesenchyme and terminal ends of the ureteric bud. Nephrons, in the different stages of development, were located around the ureteric bud which branched in the cortical nephrogenic zone and induced nephrogenesis. More mature nephrons were located in the deeper part of the cortex, close to the juxta-medullary junction. During gestation, nephrogenesis continually advanced, and the number of nephrons increased. Glomeruli changed their size and shape, while the tubules changed their length and convolution. Renal cortex became wider and contained the more mature glomeruli and the more convoluted tubules. The volume density of the tubular component of the nephron increased continually from 10.53% (LM IVa) to 27.7% (LM X). Renal corpuscles changed their volume density irregularly during gestation, increasing from 13% (LM IVa) to 15.5% (LM IVb). During the increase of gestational age, the volume density of corpuscular component of the nephron decreased to 11.7% (LM VIII), then went on increasing until the end of the intrauterine development (LM X) when corpuscles occupied 16.73% of the cortical volume. The volume density of the developing nephrons (corpuscular and tubular portion) showed the significant positive correlation (r = 0.85; p<0.01) with gestational age. CONCLUSION: The present study was one of few quantitative studies of the human developing nephron. Knowledge about the normal development of the human kidney should be important for the future medical practice. PMID- 15889593 TI - [Effects of orthodontic treatment after adenoidectomy on jaw relations in the sagittal plane]. AB - AIM: To evaluate the orthodontic treatment effects, performed immediately after adenoidectomy, on lips relation, overjet and class (Angle). METHODS: Three groups of patients, aged 6-17 years, were observed: group K--patients with oral respiration caused by adenoidal enlargement; group 1--patients with adenoidectomy performed five and more years previously; group 2--patients with one year of orthodontic treatment performed immediately after adenoidectomy. Lips and overjet status and class (Angle) was measured in the examined patients. RESULTS: In all of the three examined groups, there was a statistically highly significant increase (p<0.01) of competent lips. In the patients of group K, the overjet values ranged from 1 mm up to 12 mm where 90% of them had overjet > 4 mm and 6% < 1 mm. The front open bite, low overbite and reversed overjet was found in the patients with low overjet in this group, as well. Overjet values in the patients of group 1 ranged from -2 mm up to 12 mm where 86% had overjet > 4 mm and 10% < 1 mm. Significantly higher increase (p<0.01) of overjet values (1-4 mm in 94% of patients) was observed in the patients of group 2 in comparison to the values found in the patients from groups 0 and 1. Class II (Angle) was recorded in 81% of the patients in group 0, and 79% in group 1, while class I (Angle) was observed 100% only in the patients of group 2. CONCLUSION: Orthodontic treatment immediately after adenoidectomy was necessary for achieving the competition lips relation, normal overjet and class I (Angle). PMID- 15889594 TI - [Influence of manufacturing procedure on stability of Unguentum contra perniones preparations]. AB - BACKGROUND: Application of various technological procedures for the manufacture of officinal preparation ointment against chilblains (Unguentum contra perniones) produce essential variations in the quality and stability of the final product. Changing the sequence of admixing active substances into the ointment base indicated the presence of incompatibility between the active substances, as well as between the active substances and the ointment base components. The aim was to examine the influence of various technological manufacturing procedures on quality and stability of the preparation. METHODS: Changes in the samples of ointments and aqueous solutions of active substances were analysed microscopically. RESULTS: Microscopic analysis of hydrosoluble active substance solutions demonstrated destruction of ichthammol, induced by an acidic medium due to the dissolution of tannin and resorcinol, which is well known from the literature. It also demonstrated the destruction of tannin and resorcinol in aqueous solution, which had not been described in the literature. Application of the prescribed procedure for the manufacturing of ointment against chilblains, conceals the incompatibility reactions due to a slow dissolution of the tannin suspended in the officinal ointment base Unguentum cera lanae. Admixture of an ready-made aqueous solution of tannin caused an instant contact between ichthammol and the acidic medium, which caused the destruction or the complete absence of the formation of emulsion droplets. The problem of incompatibility reactions between lanolin alcohols in the ointment base and ichthammol and resorcinol, as well as the reaction between Peruvian balsam and sulfur from the ichthammol sulfate ion was observed. Numerous incompatibility reactions of ointment against chilblains, components indicated that it was necessary to introduce alterations in both the qualitative and quantitative composition of the preparation. CONCLUSION: Excipient preparation procedure, as well as the technological procedure for incorporation of active ingredients can cause the incompatibilities and therefore the formulation stability problems. PMID- 15889595 TI - [Quality of life in patients with allergic rhinitis and bronchial asthma]. PMID- 15889596 TI - [Gene therapy for atherosclerosis]. PMID- 15889597 TI - [Use of thoracic bioelectric impedance in the assessment of hemodynamic parameters of the cardiovascular system]. PMID- 15889598 TI - [Innominate artery war injury]. AB - AIM: A case is reported of successfully surgically treated explosive war injury to the innominate artery. CASE REPORT: A 26-year-old soldier was injured in combat by a fragment of mortar shell. In the field hospital, the wound gauze packing was applied, followed by orotracheal intubation and thoracic drainage. The soldier was admitted to MMA six hours later. Physical examination, on admission, revealed huge swelling of the neck, the absence of pulse in the right arm and the right common carotid artery. Chest x-ray revealed hemopneumothorax of the right side and the foreign metal body in the projection of the right sternoclavicular joint. Due to the suspicion of large vessel injury, a median sternotomy was immediately performed. Surgery revealed disrupted bifurcation of the right innominate artery, so the ligation was performed. Aortography was performed postoperatively, followed by the reconstruction of innominate bifurcation with synthetic grafts. Control aortography showed good graft patency, and the patient was discharged from the hospital in good general condition with palpable pulses and mild anisocoria as a sole neurological sequela. CONCLUSION: A rare and life-threatening injury was successfully managed, mainly due to the rational treatment carried out in the field hospital that helped the injured to survive and arrive to the institution capable of performing the most sophisticated diagnostic and therapeutic procedures. PMID- 15889599 TI - [Extended ankle and foot fasciotomy as an enhancement to the surgical treatment of patients with prolonged ischemia of the lower extremities]. AB - AIM: To present the technique and efficacy of extended ankle and foot fasciotomy, as a surgical limb-salvage procedure associated with the successful revascularization after the gold interval. METHOD: A retrospective review of six patients at the age of 16- 79 years (mean 39) in the period from 1996 to 2003, treated with the extended ankle and foot fasciotomy. There were four males undergoing the delayed treatment of war injuries to the lower extremity, and two females with atherosclerotic occlusive disease (a. illiacae dex and a. poplitealis dex.). The average ishemic time in the wounded patients was 22 hours (range, 14-30 hours), and ischemic time in occlusive disease (range, 48-72 hours). Clinical signs, indications, surgical access and the technique of extended ankle and foot fasciotomy was reported. RESULTS: The obtained results were classified as early and late. There was no cases with lethal outcome. Incisional wounds closed 15 days after the intervention, using secondary suture or skin transplant sec. Thiersch. The satisfactory functional results were achieved in 4 of the patients, 1 had a small edema, while amputation was performed in 1 patient. CONCLUSION: In cases of prolonged foot ischemia with edema and paresthesia in the toes, extended ankle and foot fasciotomy should be performed as a surgical limb-salvage procedure. PMID- 15889600 TI - [Dual chamber pacemaker in the treatment of paroxysmal atrial fibrillation]. AB - BACKGROUND: Atrial fibrillation is the most frequent cardiac dysrhythmia. The aim of this study was to show the role and the efficacy of a dual chamber pacemaker with the algorithm of atrial dynamic overdrive, in the suppression of paroxysmal atrial fibrillation. CASE REPORT: A woman with a classical bradycardia tachycardia syndrome, and frequent attacks of atrial fibrillation, underwent the implantation of a single chamber permanent pacemaker (WI). Pacemaker successfully treated the episodes of symptomatic bradycardia, but the patient had frequent attacks of atrial fibrillation, despite the use of different antiarrhythmic drugs, which she did not tolerate well. The decision was made to reimplant a permanent dual chamber pacemaker with the algorithm of atrial dynamic overdrive. The pacemaker was programmed to the basic rate of 75/min, while rate at rest was 55/min. In addition, sotalol was administered. After three months, the patient became asymptomatic with only 4 short-term episodes of atrial fibrillation, and a high level of atrial pacing (99%). CONCLUSION: In selected patients with bradycardia-tachycardia syndrome, atrial-based pacing seemed to be very effective in reducing the incidence of paroxysmal atrial fibrillation. PMID- 15889601 TI - [A model for therapeutic interventions in emotional responses to enforced geographical relocation of populations in Israel]. AB - BACKGROUND: The planned mandatory geographical relocation of the population inhabiting the Gush Katif settlements, planned to take place in July 2005, stands at the focus of public debate and has aroused virulent disagreements an ideological, political, national and security issues. AIMS: In order to prepare for professional assistance in coping with possible emotional problems arising from the forced relocation and in order to create support systems for professional interventions on the personal and family levels, the national health administration is seeking to design an adequate inclusive program, based at least partly on lessons learned from the relocation of populations from Sinai peninsula (Yamit and Rafiah areas) in April of 1982. However, little has been reported in the literature which can aid in designing a suitable intervention, tailored for an event such as this. Based on an analysis of the differences between combat- and terror-related trauma, and the forthcoming forced relocation of the civilian population of an entire geographical area, we propose a specifically adapted approach to professional intervention in the possible emotional consequences of the events. THE PROPOSED MODEL: The proposed principles are a mirror image of the accepted principles for combat reaction: distance from the arena as opposed to proximity, delayed intervention (after the acute phase) instead of immediacy, and expectation for actions related to adapting to a new existential reality rather than the expectation for speedy return to the arena. These principles seem to us far more suited to the demands of the complex situation ahead, in order to provide relief and assist in adaptation, and to prevent later complications of possible emotional sequelae. PMID- 15889602 TI - [Spontaneous umbilical cord hematoma]. AB - BACKGROUND: Spontaneous umbilical cord hematoma is rare and might be associated with fetal death. CASE: An emergency cesarean section was performed on a woman during the 41st week of gestation due to fetal distress. A stretched umbilical cord with hematoma was evident. The infant had tachypnea without any other physical or neurological damage. We speculate that traction of the cord led to a tear of an umbilical vessel, cord hematoma, compression of cord vessels and fetal compromise. CONCLUSION: In case of early prenatal diagnosis of spontaneous umbilical hematoma accompanied with signs of fetal distress, labor should be considered. PMID- 15889603 TI - [Eye injuries caused by rotating wire brushes]. AB - BACKGROUND: Blunt or penetrating eye injuries are quite common in industry. Workers with high risk of eye trauma do not always wear safety goggles. AIM: This is a report on three patients with wire brush eye injury who presented in our department over a 4 month period. RESULTS: In all 3 cases, the wire brush penetrated the cornea, the iris and the lens. In one case, an endophthalmitis developed, and despite appropriate treatment (including surgeries), the final visual outcome was poor. CONCLUSIONS: Eye injuries from rotating wire brushes should be treated with caution, as there is a definite risk of developing endophthalmitis. Safety goggles are recommended for the prevention of such injuries. PMID- 15889604 TI - [Transient lower limbs paralysis--a rare presenting symptom of Sjogren's syndrome]. AB - Sjogren's syndrome is commonly presented with eye and mouth dryness caused by an autoimmune destruction of lacrimal and salivary glands. Kidney involvement is not common but may appear in patients with long standing disease. We present a patient with Sjogren's syndrome whose first manifestation of the disease was related to type 1 renal tubular acidosis (RTA) causing severe hypokalemia with lower limb paralysis. The uneven distribution of limb weakness necessitated the exclusion of cord compression. PMID- 15889605 TI - [Delayed diagnosis of carcinoma of the oral cavity]. AB - Approximately 4% of cancers are located in the mouth and pharynx, 95% of which are squamous cell carcinoma (SCC). The case fatality rate of SCC is very high 50%, but early diagnosis significantly lowers the fatality rate. Diagnosis and treatment of oral lesions are shared by numerous specialists including dermatologists, otolaryngeologists, family physicians, general dentists and specialists in oral medicine and in oral surgery. The article describes a patient who was referred to a dermatologist because of a lesion of two months duration on the lateral aspects of the tongue. The dermatologist clinically diagnosed an apthous ulcer and treated it as such. When he returned after two and a half months after no clinical improvement he was diagnosed as suffering from candidiasis and treated accordingly. The unsatisfied patient turned to another dermatologist who referred him to a specialist in oral surgery. He immediately performed a biopsy. SSC was diagnosed, one third of the tongue was resected as well as the submandibular lymph nodes and salivary gland. An Israeli study has demonstrated that dentists performed better than physicians from general medical specialist fields in diagnosing early stage oral cancer. The authors suggest that patients suffering from oral lesions should be referred to specialists in oral medicine or in oral surgery for the diagnosis of oral lesions in order to diagnose oral cancer earlier. PMID- 15889606 TI - [Treatment of vaginismus with the Paula Garburg sphincter muscle exercises]. AB - OBJECTIVE: To prospectively assess and compare a novel approach to the treatment of vaginismus based on Paula Garburg's sphincter muscle exercises to the traditional approach based on Masters & Johnson. PATIENTS AND METHODS: The study population included 60 consecutive patients with vaginismus who were referred to our sex therapy clinic. The patients were randomly assigned into two groups. The study group (30 patients) underwent a step-by-step program consisting of the performance of the Paula Garburg sphincter muscle exercises to achieve pelvic muscle relaxation combined with the active introduction of Young vaginal dilators of graduated size. A control group (30 patients) underwent a behavioral treatment based on Masters and Johnson's desensitization approach that was combined with the active introduction of Young vaginal dilators of graduated size. RESULTS: All patients completed the program, all with a successful outcome, as defined by the achievement of full intercourse or the introduction of a number 6 Young vaginal dilator. The mean number of treatment sessions required for successful outcome was shorter in the study as compared to the control group (4.9 +/- 1.4 vs. 7.4 +/ 3.5, respectively. P < 0.001). Furthermore, while the percentage of patients undergoing up to 5 sessions until successful outcome was significantly higher in the study as compared to the control group (p < 0.05), the percentage of patients undergoing > 10 sessions was significantly lower. CONCLUSION: Paula Garburg's sphincter muscle exercises together with the use of Young vaginal dilators is a simple, easy and very effective treatment for vaginismus. The outcome matched or even surpassed that of the traditional behavioral approaches. PMID- 15889607 TI - [Depersonalization after withdrawal from cannabis usage]. AB - The phenomenon of depersonalization during cannabis usage (intoxication) is commonly known. However, its appearance after drug stoppage is relatively unknown. This article reviews the literature on depersonalization after cannabis withdrawal and discusses three representing cases demonstrating the severity of the problem. Clinical features are described as well as effects on functioning and the long-term nature of this disorder. The treatment approach in each case is also presented. PMID- 15889608 TI - [Post exposure prophylaxis (PEP) to prevent HIV after sexual exposure in Israel]. AB - In Israel, national guidelines exist for the management of non-occupational exposures to HIV, and post-exposure prophylaxis (PEP) is the standard of care when the risk for contracting the virus is significant. Since the efficacy, safety and cost-effectiveness of PEP after sexual exposure are not well established it is very difficult to calculate when the risk of contracting the virus is significant enough to begin therapy. Epidemiological data regarding the prevalence of HIV in specific sub-populations will help calculate this risk and facilitate decision making. Conflicting attitudes and practices exist among physicians in Israel and hopefully, the dissemination of the national guidelines will help to develop some consistency among physicians. PMID- 15889609 TI - [Antiviral prophylaxis following possible sexual exposure to HIV]. AB - In Israel, as in certain European countries, national health guidelines exist for preventive treatment after possible post-coital exposure to the HIV virus. Scientifically, this treatment is still regarded as experimental, though over the past years there is a mounting level of evidence of its efficacy. The administration of prophylaxis for HIV after unprotected coitus may be justified by the similar risk of HIV transmission for certain sexual and occupational exposures. Other considerations include: biological plausibility, the effectiveness of post-exposure prophylaxis in animal studies and occupational exposures in humans, efficacy in the prevention of vertical HIV transmission, and cost effectiveness studies. The preventive post-exposure treatment bears many side effects and possible complications, and therefore, each case should be cautiously analyzed before starting such treatment. This article reviews the scientific evidence on this topic, the considerations taken by the physician in initiating post coital HIV exposure treatment, and the guidelines available in Israel and throughout the world. PMID- 15889610 TI - [Cadherins in malignancies of the female genital tract]. AB - Cadherins are a superfamily of adhesion molecules that mediate Ca++ -dependent cell-cell adhesion necessary for normal morphogenesis and maintenance of tissue integrity. A classical cadherin molecule, such as E-cadherin, is a glycoprotein made up of three parts: an extracellular portion composed of five identical domains, a transmembrane portion composed of a single domain and a cytoplasmic portion composed of two domains. The cytoplasmic portion is anchored by means of cytoplasmic catenins to the cytoskeleton. The three amino acids sequence, histidine, alanine and valine (HAV motif) located at the most external domain of the extracellular portion, plays a key role in homophilic recognition between two cadherin molecules and cell-cell adhesion. Loss of cell-cell adhesion may be a prerequisite for malignant transformation and the invasive behavior of malignant tumors. Research of cadherin in malignancies has attracted much attention since cadherins may be proven to be reliable markers of biological behavior and prognosis The studies on cadherin in malignancies of the female genital tract have shown the following results: 1) in malignant transformation of the ovarian surface epithelium (OSE) and in epithelial ovarian carcinoma confined to the ovary (Stage I) there is a switch from N-cadherin expression to E-cadherin expression; 2) In advanced-stage epithelial ovarian carcinoma (Stages II-IV) the results are at odds: some investigators have shown a loss of E-cadherin expression most often because of hypermethylation of the promoter region of the gene, while others have demonstrated an increase in E-cadherin expression; 3) In endometrial carcinoma, E-cadherin expression is decreasing and P-cadherin expression is increasing with worsening of histologic type and differentiation, increased penetration into the myometrium, spread beyond the uterus and involvement of pelvic lymph nodes; 4) In squamous cell carcinoma of the uterine cervix E-cadherin expression is decreasing with tumor progression and in adenocarcinoma of the uterine cervix P-cadherin expression is increasing with tumor progression. It is hoped that the development of drugs that amend cell-cell adhesion will improve the prognosis of patients in whom tumor progression is associated with decrease or loss of cadherin expression. PMID- 15889611 TI - [Medical management in the chemical terrorism scene]. AB - The Tokyo subway sarin attack in March 1995 demonstrated the importance of preparedness toward a chemical terrorist attack. Emergency medical teams on the scene are valuable, beside the medical treatment of casualties, in the cognition of toxicant involvement and later in the recognition of the specific toxidrome involved. The chemical terrorism scene is a contaminated area; therefore, first responders have to be protected from both percutaneous and inhalational exposure to toxic materials. This protection is also against secondary evaporation (gas off) from contaminated casualty, hence the importance of disrobing casualties on the scene as soon as possible. Once the recognition of toxicological involvement have been made, the next crucial decision is whether the clinical toxidrome is of cholinergic toxicity (e.g. organophosphate or carbamate intoxication) in which there are automatic injectors for treatment available on the scene, or any other toxidrome (such as irritation or vesicants) in which, beside general measures, like oxygen delivery and airway support, there is not a specific antidotal treatment on the scene. The clinical detection and identification of the chemical toxidrome involved is of utmost importance since it promotes the antidotal treatment quickly and efficiently. The key to the medical management of such events is based on decisions that have to be taken as soon as possible according to the clinical judgment of medical teams on the scene. PMID- 15889612 TI - [Sexually transmitted diseases--an update]. AB - Sexually transmitted infections represent an worldwide challenge for the public health. According to WHO estimates, approximately 330 million people are infected annually by curable sexually transmitted infections (AIDS excluded). and their incidence has been increasing, particularly in high-risk populations. Like in other developed countries, the occurrence of venereal diseases in Israel has been increasing recently. In addition, a sharp rise has been observed in the resistance rate of gonococci to fluoroquinolones. The purpose of the present review is to update the information on the epidemiology, diagnosis, therapy and prevention of sexually transmitted diseases (STDs) in Israel. In response to the reemergence of these diseases in Israel, it was decided in the Ministry of Health to open for the first time ever STD clinics in the two cities with the highest disease burden, namely Tel Aviv and Haifa. These clinics are staffed with a multidisciplinary group of specialists, including gynecologists, dermatologists, epidemiologists, nurses and social workers. PMID- 15889613 TI - [Degenerative lumbar spinal stenosis--review of the current diagnosis and treatment]. AB - Degenerative lumbar spinal stenosis (LSS) is a common cause of disabling back and leg pain, and spinal claudication among older persons. As the population grows older this condition is encountered more frequently. The degeneration of the intervertebral discs and the facet joints results in the narrowing of the spinal canal and neural foramina. Non-operative management is often of limited value and includes physical therapy, anti-inflammatory medications, and the injection of epidural steroids. The condition of a substantial portion of the patients does not deteriorate and remains unchanged. If non-operative treatment fails, surgical treatment should be considered. Surgical success rates as high as 90% have been reported. Few prospective studies have shown an advantage of operative treatment over non-operative treatment. A controversy exists pertaining to whether results in patients who have had surgery after failure of non-operative treatment were less satisfactory than results in patients who were operated at baseline. More randomized prospective studies are needed to obtain clear-cut conclusions regarding this controversy, and other controversies such as the exact role of surgical treatment. PMID- 15889614 TI - [Pharmacotherapy in schizophrenia--comparison of second generation antipsychotic agents]. AB - Schizophrenia is a severely limiting chronic illness. The disease is characterized by periods of psychoses, remission, and sometimes even recovery. Effective pharmacotherapy for schizophrenia began with the development of antipsychotic neuroleptics in the 1950's, followed by the second generation of antipsychotic agents developed in the 1990's. Across time, treatment with some of these preparations revealed extrapyramidal and other side effects, including metabolic phenomenon, such as: an increase in the incidence of diabetes, and weight gain. When choosing the appropriate medication from among the second generation of antipsychotic agents, the following aspects should be considered; cost, efficacy, unique advantages, employment rehabilitation, relapse and metabolic side effects. Therapeutic algorithms and recommended guidelines are helpful when planning treatment strategies. Information regarding prior beneficial treatments, cost, and the patient's quality of life should be considered. Today, when patients' rights, economic and legal aspects are emphasized, the physician must take into account multiple factors when choosing the most appropriate treatment for the patient. PMID- 15889615 TI - [Medicine in the concentration camps of the Third Reich]. AB - Between 1942 and 1944 millions of prisoners were subjected to forced labor in concentration camps throughout the Third Reich, all the while being the victims of a systematic and "scientific" extermination policy. Though the policy was directed mainly against Jews, it was implemented against other "inferior races" as well. The prisoners, stripped of all rights, experienced constant humiliation, uncertain survival and terror. The harsh living condition, characterized by crowding, absent sanitation and poor personal hygiene led to considerable morbidity, mainly due to injuries, infectious diseases and famine, and to high mortality rates. Medical care in the camps was the responsibility of the S.S. Each camp had a chief S.S. physician accompanied by a number of assistants and orderlies. There was also a parallel system of "prisoner-physicians." There was a chief prisoner-physician in every camp, and each block was assigned a "block doctor" who was responsible for sanitation, the removal of corpses, setting up the sick- call and authorizing sick leave. Work teams were accompanied by "mobile doctors" (Streckenpfleger), who dispensed first aid for work injuries. Prisoner physicians were also charged with disinfecting the blocks and maintaining hygienic conditions in the camp. Every camp had one or more blocks, called "Reviers", that were used for treatment and hospitalization. In the larger camps a number of blocks were designated to function as a sort of hospital (Krankenbau). At times one camp out of a group of camps would be set aside as quarantine, primarily for patients with infectious diseases. Officially, the "Revier" was the responsibility of an S.S. physicians', assisted by a chief prisoner-physician, his assistants and, at times, nurses. But in actuality the Reviers were managed by prisoners (Capos) who did not have medical training but were authorized to make decisions in medical matters such as operations and, on occasion, even performed them. The Reviers suffered from shortage of everything: beds, manpower, diagnostic and therapeutic means, sterile supplies and drugs for anaesthesia. Crowding was the rule, and each cot was occupied simultaneously by several patients regardless of their condition or the nature of their disease. Life-saving procedures could not be performed and surgery, if done, was usually without anaesthesia. Mortality was high, and the Revier was commonly perceived by the prisoners as a place of death, not of healing. Occasionally there were visits, or inspections, by the S.S. physicians. The main purpose of those visits was "selection". Patients with no chance of recovery were sent to their death and the others were returned to work in spite of their condition. The prisoner physicians made every effort to prevent this "selection" in various ways. They were often punished or even executed for their efforts. PMID- 15889616 TI - [The history of blood transfusions]. AB - Numerous lives are being saved each passing day via blood transfusions. Merely a century ago, blood transfusions were considered dangerous. Yet, for physicians of the 21st century, the absence of blood transfusions is unimaginable. The history of blood transfusion travels through states, religions and discoverers--from a pope to a French charlatan and from an English doctor to an Austrian converted Jewish physician. This article attempts to shade light upon some of the central events concerning the history of blood transfusions. PMID- 15889617 TI - Expression of ubiquitin-related enzymes in the suprachiasmatic nucleus with special reference to ubiquitin carboxy-terminal hydrolase UchL1. AB - There is growing evidence that ubiquitin-proteasome system plays an important role for the generation of circadian rhythms in mice as in Drosophila. Here we examined the expression of ubiquitin-related enzymes (Ubce5, UbcM4, Ube2v, Ube2d2, UchL1, UchL3, Ubp41, UfdlL, beta-TrCP) in the suprachiasmatic nucleus (SCN). At mRNA level, the-expression of these enzymes were faint to moderate except ubiquitin carboxy-terminal hydrolase L1 (UchL1), a dominant deubiquitinating salvaging enzyme. Although strongly expressed in the SCN, UchL1 mRNA did not show the rhythm in the SCN in both light-dark and constant dark conditions. PMID- 15889618 TI - Bone formation in beta-tricalcium phosphate-filled bone defects of the rat femur: morphometric analysis and expression of bone related protein mRNA. AB - The purpose of the current study was to evaluate the bone formation when beta tricalcium phosphate (TCP) was implanted in bone defects of rat femurs. beta-TCP granules were applied to defects created in the femurs of 65 male rats who were sacrificed 3, 7, 10, 14 or 30 days later. Bone tissues were embedded in paraffin, serial sections were cut and then stained with hematoxylin-eosin. Histomorphometric analyses were also conducted. Furthermore, total mRNAs were extracted, homogenized, and reverse transcribed, after which quantitative PCR assays were conducted with a LightCycler using the double-stranded DNA dye Syber Green I with primers for either rat osteopontin or osteocalcin. Tissues in defects without beta-TCP were used as controls. The amount of newly formed bone tissue in the beta-TCP implanted group was significantly greater in both the side areas and the central area of defects than in the control group. Expressions of osteopontin and osteocalcin mRNAs of cells in the defects of the experimental group were up-regulated compared with the control group at all time periods. Taken together, these results prove that beta-TCP is an appropriate material for osteoconduction and promotes bone formation in bone defects. PMID- 15889619 TI - Effects of stretching stress on the muscle contraction proteins of skeletal muscle myoblasts. AB - Several studies have reported that growth and differentiation of cultured myoblasts can be facilitated by applying appropriate mechanical stimulus. However, the effects of mechanical stimulus on the characteristics of muscle fibers have not yet been fully elucidated. In this study, we gave mechanical stress to C2C12 cells, which were myoblasts derived from mice skeletal muscle. The following myosin heavy chain (MHC) isoforms were investigated in order to clarify muscle characteristics: MHC-2b, 2d and 2a, all of which are fast-twitch fibers. After inoculating cells on a silicone chamber, the chamber was mechanically stretched, and a LightCycler was used to measure the mRNA expression of each MHC isoform at several times. The results showed that, with mechanical stretching, the expression of MHC-2b was initially high. On the other hand, without stretching, the expression of MHC-2d increased over time, but with stretching, it was hardly seen. Furthermore, the expression of MHC-2a was significantly high in the stretching group. These results of this study suggest that, when intermittently stimulated, myoblasts express increased levels of MHC 2a isoform. Therefore, it is indicated that myocytes respond to environmental changes not only to facilitate growth and differentiation, but also to alter muscle function actively at the MHC isoform level. PMID- 15889620 TI - Periodontal regeneration in intrabony defects after application of enamel matrix proteins with guided tissue regeneration: an experimental study in dogs. AB - The aim of this study was to evaluate the effects of enamel matrix proteins (EMP) at the early stage of wound healing in the periodontal tissues by a combination treatment with guided tissue regeneration (GTR). Intrabony defects were produced surgically at the distal aspects of both mandibles in six beagle dogs. At 12 weeks following the surgery, the defects were exposed using a full thickness mucoperiosteal flap procedure. Subsequently, the defects were treated by the following treatments: a control group treated with GTR alone, and an experimental group treated with a combination of GTR and EMP. After one, two, four and eight weeks of the treatment, the animals were sacrificed, and sections of the tissue were stained and evaluated microscopically. After one and two weeks, the proliferating cell nuclear antigen (PCNA)-positive cell ratios of the experimental group were significantly greater than that of the control group. After 2 and 4 weeks, new bone and new cementum formation in the experimental group were significantly greater than those in the control group (P < 0.05). However, after 8 weeks, no statistical difference was found between the two groups in new bone or cementum formation. The study results suggest that a maturation of periodontal ligament cells might contribute, during the early stage of periodontal healing, to stimulate a proliferation of periodontal ligament cells. PMID- 15889621 TI - Exercise-related novel gene is involved in myoblast differentiation. AB - In this study we tried to identify new genes or proteins in skeletal muscle induced by exercise. We analyzed alterations of protein expression in mouse gastrocnemius muscles induced by swim-exercise using two dimensional gel electrophoresis and mass spectrometry. Nine spots were significantly altered between control and swim groups. One of the four protein spots whose expression was decreased was identified as functionally unknown "expressed sequence AI854635" gene. The AI854635 gene has C2H2 type zinc finger motif, and is considered to be a transcription factor. The mRNA of AI854635 gene was expressed in skeletal muscle, brain, kidney, and thymus. To elucidate the function of the AI854635 gene we analyzed mRNA expression levels during C2C12 myoblast differentiation. C2C12 myoblast began to form myotube around 20 h after the initiation of differentiation. The mRNA expression levels of AI854635 gene was significantly induced around 6 h and increased till 48 h, indicating a pivotal role in myoblast differentiation. Although the significance of decreased expression of AI854635 gene induced by swim-exercise is not clear, we found that this gene is involved in myoblast differentiation. PMID- 15889622 TI - Influence of prostaglandin A2 and 2-methoxyestradiol on Bax and Bcl-2 expression levels in cervical carcinoma cells. AB - Proteins of the Bcl-2 family are key regulators of apoptosis. Bax can be regarded as pro-apoptotic, whereas Bcl-2 is perceived as anti-apoptotic. It has been proposed that an increased ratio of pro-apoptotic Bax to anti-apoptotic Bcl-2 can be associated with apoptosis. Since prostaglandin A2 (PGA2) and 2 methoxyestradiol (2-ME) play an active role in the induction of apoptosis, the influence of 20 microg/ml PGA2 and 1 microM 2-ME was investigated on Bax and Bcl 2 expression levels in cervical carcinoma cells. Both PGA2 and 2-ME exposure led to statistically significant increases in Bax expression levels. Cells were shown to be more susceptible to the effects of 2-ME than to the effects caused by PGA2. In contrast, no statistically significant effects were observed on Bcl-2 expression levels after exposure to PGA2 and 2-ME. The Bax/Bcl-2 ratios for PGA2- and 2-ME-exposed cells were 2.06 and 1.87 respectively, normalised against Bcl-2 levels. Further investigation of the function and regulation of the Bcl-2 family will allow researchers to consider potential pathways of apoptosis signaling mechanisms for diseases where apoptosis can potentially be controlled. PMID- 15889623 TI - Individual, family, school, and community risk and protective factors for depressive symptoms in adolescents: a comparison of risk profiles for substance use and depressive symptoms. AB - This study examines the relationship between adolescent depressive symptoms and risk and protective factors identified for substance use. A questionnaire, developed to measure these factors in a young person's community, family, school, peer group, and individual characteristics for substance use, was used to assess associations with self-reported depressive symptoms. Data were provided by a representative sample of 8984 secondary school students in Victoria, Australia. The prevalence of depressive symptoms was 10.5% (95% CI 9.2,12.0) for males and 21.7% (95% CI 20.3,23.7) for females. Depressive symptoms were associated with factors in all domains, with the strongest associations in the family domain. Strong relationships were found between the number of elevated risk and protective factors and depressive symptoms, maintained after adjusting for substance use. Patterns of associations were similar for users and nonsubstance users. The findings indicate that prevention programs targeting factors for substance use have the potential to impact on depression. PMID- 15889624 TI - Effects of presenting heavy drinking norms on adolescents' prevalence estimates, evaluative judgments, and perceived standards. AB - Correcting normative information about the prevalence of heavy drinking is a key element in many prevention programs. To isolate the influence of normative information on older high school students' (n = 230) alcohol-related judgments, the effects of delivering normative information in different contexts (no normative information, normative information only, normative information plus a self-focusing comparison to one's drinking) and under different measurement conditions (public, private) were examined. First, relative to presenting no norms, presenting norms both with and without a self-focus reduced the underestimation of the percent of high school students who never drink heavily. Second, effects on both positive and negative evaluations of heavy drinking were examined independently. Heavy drinking students more strongly endorsed positive evaluations of heavy drinking than did non-heavy drinking students, but this self serving bias was limited to the normative information only condition. Normative information failed to impact negative evaluations of heavy drinking for students at all drinking levels. Third, in judging the acceptable number of heavy drinking days approved by others, presenting the normative information in both contexts (relative to presenting no norms) led to more conservative judgments. Yet, only the normative context that added a self-focus to the norm led students to adopt more conservative personal standards for the acceptable number of heavy drinking days. Finally, public versus private measurement did not affect any of the dependent variables. The findings are discussed as they relate to confrontational versus empathic styles in delivering interventions. PMID- 15889625 TI - Intraindividual variability of school bonding and adolescents' beliefs about the effect of substance use on future aspirations. AB - The study examines the dynamic relationship between school bonding, beliefs about the deleterious effects of substance use on future aspirations, and subsequent substance use among a sample of 1065 male and female middle school students. First, a mediation model was assessed. Adolescents' perceptions about the harmful effects of substance use on their future aspirations emerged as a salient mediator of the relationship between school bonding and subsequent substance use. Second, the intraindividual variability of school bonding and its effect on students' beliefs about the potential harm of substance use on future aspirations was assessed through random-coefficient models. Students who tended to be poorly bonded to school were less likely to perceive that substance use may impede the attainment of their future goals. Furthermore, a strong intraindividual effect of school bonding was observed, indicating that as a student became more or less bonded to school his/her belief that substance use could affect future aspirations similarly changed. PMID- 15889626 TI - Schools and homes in partnership (SHIP): long-term effects of a preventive intervention focused on social behavior and reading skill in early elementary school. AB - This paper reports a randomized controlled trial of the effects of behavioral parenting skills training, social skills training, and supplemental reading instruction on the social behavior of early elementary school children (K through 3). We selected children based on teacher-rated aggressive behavior or reading skill deficits, delivered the intervention over a 2-year period, and obtained follow-up data for two additional years. The intervention affected only two of eight measures of child functioning--parent daily reports of antisocial behavior and parent ratings of coercive behavior. There was evidence that parents of boys in the intervention condition displayed significantly greater declines in their rated use of coercive discipline than did parents of boys in the control condition. PMID- 15889627 TI - Ethnic differences in relationships between risk factors and adolescent binge drinking: a national study. AB - This study examines ethnic differences in relationships between a large number of risk factors and adolescent binge drinking with data collected from 14 to 17 year olds who participated in the 1999 National Household Survey on Drug Abuse (NHSDA). Logistic regression analyses were conducted to determine whether 39 risk factors in community, family, school, and peer-individual domains were differentially associated with past-30-day binge drinking among youth in ethnic minority groups (black, Hispanic and Asian) relative to whites. Forty-three percent (17) of the risk factors examined were differentially associated with binge drinking in at least one of the ethnic groups relative to whites. Most of these risk factors were more strongly associated with binge drinking among white adolescents than Hispanic and black youth, but not Asians. The direction of the relationships between these risk factors and binge drinking, however, was usually the same for whites and ethnic minority groups. A multivariate prediction model indicated that risk factors in the community, family, school, and peer-individual domains accounted for 27% of the variance in binge drinking for white adolescents, 22% for Hispanics, 10% for blacks, and 39% for Asians. These findings suggest that research is needed to identify additional risk factors that are associated with binge drinking among adolescents, particularly blacks, Hispanics and possibly other ethnic minority groups. PMID- 15889628 TI - Acculturation, income, education, safety belt use, and fatal motor vehicle crashes in California. AB - This paper investigates the role that acculturation, income, and education play in safety belt nonuse among Californian drivers involved in fatal Motor Vehicle Crashes (MVCs). To achieve this goal, measures of acculturation, income, and education were stochastically incorporated into the Fatality Analysis Reporting System (FARS). Using the 1990 California Tobacco Survey and U.S. Census data, we estimated the combination of zip-code-based measures that most accurately predicts an individual, language-based acculturation index for Hispanics and Asians. Logistic regression was used to investigate the role of these variables in safety belt nonuse in fatal MVCs. We found that acculturation has a positive direct effect on safety belt use among Hispanics. We hypothesize that this positive direct effect is caused by Hispanic immigrants learning the benefits of wearing safety belts. However, our study also suggests an indirect negative effect of acculturation on safety belt use through drinking and driving. Prevention programs aimed at increasing the safety of Hispanic drivers not only need to take acculturation differences into account, but also need to be comprehensive in their message, simultaneously targeting both seat belt nonuse and drinking-and-driving problems. PMID- 15889629 TI - Fallon, UMass partner to create regional EMR network. PMID- 15889630 TI - Capitated organizations should reassess their risk coverage, say experts. AB - Capitated organizations should reassess their risk coverage, say experts. With all signs pointing to a growth spurt in capitation during 2005, Charles Crispin, president of Evergreen Re, a Stuart, FL-based reinsurance brokerage and consulting company, says capitated physician organizations need to reassess their carve-out strategy and make sure they have enough reinsurance to avoid catastrophic loss. PMID- 15889631 TI - Study: hospitalizations increase when only generic drugs are used for Medicare Advantage members. AB - Hospitalizations increase when only generic drugs are used for Medicare Advantage members. A Medicare health plan in California that offered generic-only prescription drug coverage reduced its own per member per month costs for pharmacy, but members paid higher out-of-pocket costs and the number of hospital admissions rose. PMID- 15889632 TI - Integrated delivery systems focus on service delivery after capitation efforts stall. AB - Integrated delivery systems focus on service delivery after capitation efforts stall. Integrated delivery systems are going through changes that are focusing the provider organizations more on delivering care than managing risk, says Dean C. Coddington, one of the leading researchers into capitated organizations and a senior consultant with McManis Consulting in Denver. PMID- 15889633 TI - [Helicobacter pylori, mycoplasmas, and gastric diseases]. PMID- 15889634 TI - [Clarification regarding the article on diarrhea and type of feeding]. PMID- 15889635 TI - [The influenza pandemic: potential impact of aviary influenza]. PMID- 15889636 TI - Clinical follow up of mexican women with early onset of breast cancer and mutations in the BRCA1 and BRCA2 genes. AB - OBJECTIVE: This study describes the presence of mutations in BRCA1 and BRCA2 genes in a group of Mexican women and the clinical evolution of early onset breast cancer (EOBC). MATERIAL AND METHODS: A prospective hospital-based study was performed in a sample of 22 women with EOBC (7 in clinical stage IIA, 8 in IIB, and 7 in IIIA). The patients attended a tertiary care hospital in northeastern Mexico in 1997 and were followed up over a 5-year period. Molecular analysis included: 1) a mutation screening by heteroduplex analysis (HA) of BRCA1 and BRCA2 genes and 2) a sequence analysis. RESULTS: Of 22 patients, 14 (63.6%) showed a variant band detected by heteroduplex analysis of the BRCA1 and BRCA2 genes: 8 polymorphisms, 4 mutations of uncertain significance, and 2 novel truncated protein mutations, one in BRCAI (exon 11, 3587delT) and the other in the BRCA2 gene (exon 11, 2664InsA). CONCLUSIONS: These findings support future studies to determine the significance and impact of the genetic factor in this Mexican women population. PMID- 15889637 TI - [Heterogeneous distribution of the prevalence of anti-Trypanosoma cruzi antibodies among blood donors in the State of Puebla, Mexico]. AB - OBJECTIVE: To determine the seroprevalence and associated factors, of antibodies against Trypanosoma cruzi (T. cruzi Ab) among blood donors living in rural and suburban areas and risk regions. MATERIAL AND METHODS: A cross-sectional study was conducted from January to December 2003, in 2489 blood donors of seven regions of Puebla, who were evaluated for mandatory viral and T. cruzi serological tests using validated procedures. RESULTS: The seroprevalence for T. cruzi Ab was 1.24% (31/2489), similar to hepatitis C (HVC) (1.5%) and higher than human immunodeficiency virus (HIV) (0.4%) and hepatitis B (HVB) (0.3%). The highest seroprevalences were observed in the regions of Tehuacan-Sierra Negra and Mixteca, up to 2.6%, while in Sierra nororiental and Angelopolis no positive blood donors were identified. A positive association was observed between seropositivity and being older than forty years and being born and raised in Tehuacan-Sierra Negra and Mixteca. CONCLUSIONS: T. cruzi seroprevalence distribution is heterogeneous, from 0% to 2.6%, with higher seroprevalences in the regions of Tehuacan-Sierra Negra and Mixteca. PMID- 15889638 TI - [Effect of a School Breakfast Program on the prevalence of obesity and cardiovascular risk factors in children]. AB - OBJECTIVE: To determine the effect of a School Breakfast Program on obesity and some cardiovascular risk factors in 6 to 10 year old schoolchildren. MATERIAL AND METHODS: A quasi-experimental prospective study was conducted in 2002-2003, in 17 municipalities of Sonora State, Mexico. The intervention group consisted of 254 children participating in a School Breakfast Program (SBP group). The control group (NSBP group, n=106) included children who did not participate in the program. In both groups the body mass index for age (BMI/age) and body composition by electrical resistance using bioelectrical bioimpedance analysis (BIA) were estimated at the beginning and at the end of a 9-month period. In a subgroup of 264 school-children (SBP and NSBP children), serum cholesterol, triglycerides and fasting glucose were measured at the start and at the end of the program. RESULTS: The body mass index in the SBP and NSBP groups was not different at the start or at the end of the school period (p>0.05). The proportion of overweight and obese children and the percentage fat remained similar in both groups. However, the lean mass increased (p>0.05) at the end of the school period in both groups, and hence, cannot be attributed to the program. The biochemical parameters showed no change (p>0.05) in blood glucose, total serum cholesterol, and triglycerides in either group at the end of the school period. CONCLUSIONS: Study results showed no evidence of a negative effect of SBP in terms of risk factors for obesity and cardiovascular disease. PMID- 15889639 TI - [Gender based violence: knowledge and attitudes of health care providers in Nicaragua]. AB - OBJECTIVE: To assess the knowledge and attitudes of health care personnel towards the identification and referral of gender-based violence victims (GBV). Also, to identify barriers to identification and referral of GBV, and to assess the levels of knowledge about Norms and Procedures for Intra-Family Violence Care by the health care personnel of the Nicaraguan's Minister of Health (MINSA, for its initials in Spanish). MATERIAL AND METHODS: A cross-sectional study was conducted among healthcare professionals, including physicians, nurses and nursing technical aides (n=213), in 5 of the 17 Local Systems of Integral Attention (SILAIS) from the Integral Program of Attention for Women, Children and Adolescence (AIMNA) in the primary level of attention in MINSA, from April to June 2003. Attitude was measured with a Likert scale and an awareness index was created for intra-family violence care guidelines. The information was obtained using a self-administered instrument, based on the questionnaire of the study made among the personnel of the Mexican Institute for Social Security (IMSS, for its initials in Spanish), Morelos, Mexico. A logistic regression model was used to evaluate the association between attitude and several factors, as well as with the knowledge of care guidelines. RESULTS: In our population, 76.06% showed an attitude opposing GBV. In the multivariate analysis, the factors associated with opposition to GBV were: medical profession (OR 6.5, 95%CI 2.70-15.82), having a middle (OR 4.3, 95%CI 1.87-10.26) or high level (OR 3.3, 95%CI 1.03-10.75) of knowledge about intra-family violence guidelines and the closeness to relatives or friends who were victims of gender violence (OR 3.2, 95%CI 1.56-6.80). The lack of training on the subject (59.9%), fear of getting involved in legal issues (52.6%), and the concept that violence is a private affair and not a social one (50.7%), constituted the most important barriers to providing medical care. CONCLUSIONS: The health care personnel generally were observed to have high values in regard to an attitude of rejection towards GBV. However, we found barriers that show persisting traditional beliefs, such as considering violence to be a personal issue. Therefore, in order, to ensure a substantial improvement, better training about this subject is needed in medical school curricula with an emphasis on the gender perspective. The finding of the present study will allow improvements in health care reforms at the first level of care in the health sector in Nicaragua. PMID- 15889640 TI - [Emotional ailments, help seeking behaviors and care expectancies in a marginal urban community]. AB - OBJECTIVE: To obtain information about emotional ailments based on the experience of adults in a socially underserved urban community, their social support resources to cope with them, and their health care expectancies. MATERIAL AND METHODS: The study began in 2000, in the Magdalena Contreras District of Mexico City. Data collection was conducted in 2001-2003, using a multimethodological approach that included in depth interviews of female and male adults (n= 16) performed during 2000-2002. A semistructured questionnaire (n=204) was applied in the first semester of 2002 and data entering was performed in the second semester of that year. Quantitative analysis was performed in 2003. RESULTS: The community informants evidenced a lack of social support networks. For women, the expected support to cope with emotional disorders should come from the nuclear family, although in practice their partner represented a source of discontent. For men, the main expected source of support was their wives. CONCLUSIONS: Based on these results, some reflections are proposed around intervention programs considering both the perspective of the community members and the multidisciplinary work. PMID- 15889641 TI - [Immunogenicity and safety of the influenza vaccine, in a population older than 55-years in Mexico]. AB - OBJECTIVE: To confirm the immunogenicity and tolerance of the inactivated, fractionated, and purified influenza vaccine, in a Mexican adult population aged 55 and older, medically served at a Petroleos Mexicanos Hospital (Pemex, Mexican Oil Company). MATERIAL AND METHODS: The study was conducted between November and December, 2000, among ninety adult subjects aged 55 years and older who were seen at the Hospital Central Sur Pemex. The primary endpoints regarding immunogenicity were the percentage of individuals with protective antibodies targeting hemagglutinins higher than or equal to 1:40, and the percentage of subjects who seroconverted as measured by a four-fold increase in protective antibody production. Secondary endpoints included the frequency of local and systemic reactions to the vaccine. An additional criterion that was evaluated included antigen-antibody affinity assays to measure the polyclonal antibody response to the vaccine and the specific generation of high-affinity antibodies to viral proteins, before and after vaccination. RESULTS: The antibody protection rate was 95.6% against the HINI strain, 98.9% against the H3N2 strain, and a 100% against the B/Yamanashi strain. Seroconversion to the HINI strain was elicited in 74.4% of subjects, to the H3N2 strain in 88.9%, and to the B/Yamanashi strain in 82.2%. Eighteen (20%) subjects developed local reactions; 17 (18.8%) developed a systemic reaction post vaccination at day 5 and nine subjects (10%) at day 28. Local reactions consisted of pain in 10 (11.1%) subjects, redness in 8 (8.8%), and induration in 6 (6.6%). General malaise, headache, and fever were identified in 10, 8.8, and 0% of subjects, respectively, at day 5, and in 4.4, 6.6, and 0%, respectively, at day 28. CONCLUSIONS: Influenza vaccine was highly immunogenic in a healthy Mexican adult population aged 55 years and older. The generation of high-affinity antibodies to the virus after vaccination was also demonstrated. Local and systemic adverse reactions to the vaccine identified in our study were similar to those in previous reports. The results of this study can be extrapolated to other health institutions serving this adult population to increase influenza vaccine coverage rates. PMID- 15889642 TI - [Distribution of Aedes albopictus (Skuse) in Nuevo Leon, Mexico, 2001-2004]. AB - OBJECTIVE: To determine the distribution of Aedes albopictus (Skuse) in Nuevo Leon between 2001 and 2004. MATERIAL AND METHODS: Standardized methods were used to collect 6371 samples of mosquito's larvae from 167 sites. The proportions of samples positive for Ae. albopictus were obtained by municipality and year, using the software program Excel. RESULTS: There were 1179 samples positive for Ae. albopictus. This species was distributed over 10 municipalities, as follows: Five on the Gulf of Mexico Coastal Plains, which predominantly have a semi-warm, sub humid climate, and precipitations almost year round (Allende, Montemorelos, Linares, General Teran, and Santiago); two (Sabinas Hidalgo, and Cadereyta) localized on the North American Great Plains, which have a warm, semiarid climate, and scarce precipitations. The final three were Apodaca, San Nicolas, and Escobedo distributed over the metropolitan area of Monterrey City. CONCLUSIONS: Up to January 2004, the second most important epidemiological vector in the transmission cycle of Ae. albopictus worldwide was found in ten municipalities of Nuevo Leon State, Mexico. PMID- 15889643 TI - [The Social Protection Health System (Seguro Popular de Salud). Seven perspectives. An interview of Salud Publica de Mexico with Rogelio Gomez Hermosillo, National Coordinator of the "Oportunidades" program]. PMID- 15889644 TI - [The Social Protection Health System (Seguro Popular de Salud). Seven perspectives. An interview of Salud Publica de Mexico with Christian Baeza, World Bank executive]. PMID- 15889645 TI - [Mexico's mortality statistics. Deaths registered in 2003]. PMID- 15889646 TI - Thermal synthesis of nucleoside H-phosphonates under mild conditions. AB - Nucleosides react rapidly with ammonium phosphite ((NH4)2HPO3) at 60 degrees C to produce good yields of nucleoside-5'-phosphite monoesters within 24 h. Under the same conditions, ammonium phosphate is unreactive, producing low yields of nucleotide only after extended reactions. These results confirm earlier suggestions that nucleoside H-phosphonates and their possible condensation products may have been produced on the primitive earth more easily than nucleotides. PMID- 15889647 TI - Synthesis of nucleoside N-phosphoamino acids and peptide formation. AB - Nucleoside N-phosphoamino acids were synthesized through Atherton-Todd reaction of nucleoside H-phosphonate with amino acids, and their structures were confirmed by NMR and ESI-MS. After nucleoside N-phosphoamino acid was incubated in anhydrous methanol at 40 degrees C for 72 h, di- to tetra-peptide derivatives were detected by ESI-MS, and their structures were further identified by multistage mass spectrometry. These and previously published studies in aqueous solution suggest that nucleoside N-phosphoamino acids could have been prebiotic precursors of oligopeptides. PMID- 15889648 TI - Sites for phosphates and iron-sulfur thiolates in the first membranes: 3 to 6 residue anion-binding motifs (nests). AB - Nests are common three to six amino acid residue motifs in proteins where successive main chain NH groups bind anionic atoms or groups. On average 8% of residues in proteins belong to nests. Nests form a key part of a number of phosphate binding sites, notably the P-loop, which is the commonest of the binding sites for the phosphates of ATP and GTP. They also occur regularly in sites that bind [Fe2S2](RS)4 [Fe3S4](RS)3and [Fe4S4](RS)4 iron-sulfur centers, which are also anionic groups. Both phosphates and iron-sulfur complexes would have occurred in the precipitates within hydrothermal vents of moderate temperature as key components of the earliest metabolism and it is likely existing organisms emerging in this milieu would have benefited from evolving molecules binding such anions. The nest conformation is favored by high proportions of glycine residues and there is evidence for glycine being the commonest amino acid during the stage of evolution when proteins were evolving so it is likely nests would have been common features in peptides occupying the membranes at the dawn of life. PMID- 15889649 TI - UV circular polarisation in star formation regions: the origin of homochirality? AB - Ultraviolet circularly polarised light has been suggested as the initial cause of the homochirality of organic molecules in terrestrial organisms, via enantiomeric selection of prebiotic molecules by asymmetric photolysis. We present a theoretical investigation of mechanisms by which ultraviolet circular polarisation may be produced in star formation regions. In the scenarios considered here, light scattering produces only a small percentage of net circular polarisation at any point in space, due to the forward throwing nature of the phase function in the ultraviolet. By contrast, dichroic extinction can produce a fairly high percentage of net circular polarisation ( approximately 10%) and may therefore play a key role in producing an enantiomeric excess. PMID- 15889650 TI - Animal nutrition with feeds from genetically modified plants. AB - Plant breeders have made and will continue to make important contributions toward meeting the need for more and better feed and food. The use of new techniques to modify the genetic makeup of plants to improve their properties has led to a new generation of crops, grains and their by-products for feed. The use of ingredients and products from genetically modified plants (GMP) in animal nutrition properly raises many questions and issues, such as the role of a nutritional assessment of the modified feed or feed additive as part of safety assessment, the possible influence of genetically modified (GM) products on animal health and product quality and the persistence of the recombinant DNA and of the 'novel' protein in the digestive tract and tissues of food-producing animals. During the last few years many studies have determined the nutrient value of GM feeds compared to their conventional counterparts and some have additionally followed the fate of DNA and novel protein. The results available to date are reassuring and reveal no significant differences in the safety and nutritional value of feedstuffs containing material derived from the so-called 1st generation of genetically modified plants (those with unchanged gross composition) in comparison with non-GM varieties. In addition, no residues of recombinant DNA or novel proteins have been found in any organ or tissue samples obtained from animals fed with GMP. These results indicate that for compositionally equivalent GMP routine-feeding studies with target species generally add little to nutritional and safety assessment. However, the strategies devised for the nutritional and safety assessment of the 1st generation products will be much more difficult to apply to 2nd generation GMP in which significant changes in constituents have been deliberately introduced (e.g., increased fatty acids or amino acids content or a reduced concentration of undesirable constituents). It is suggested that studies made with animals will play a much more important role in insuring the safety of these 2nd generation constructs. PMID- 15889651 TI - Effects of dietary conjugated linoleic acid supplementation on performance and immune function of weaned pigs. AB - Two experiments were conducted to evaluate the effect of dietary conjugated linoleic acid (CLA) on performance and immune responses of weaned pigs. In Exp. I, 72 crossbred pigs weaned at 19 to 23 days of age and weighing 7.20 +/- 0.11 kg were randomly allotted to four diets supplemented with CLA at 0, 1, 2 or 3%. On day 14, pigs were injected with ovalbumin (1mg per kg BW) and blood samples were collected on day 7 and 14 after injection to test the specific OVA antibody. In Exp. II, 36 crossbred pigs weaned at 26 to 30 days of age and weighing 8.12 +/- 0.14 kg were randomly divided into two diets containing either 0 or 2% CLA. On day 14 and 28, blood samples were obtained to determine the lymphocyte proliferation and PGE2 levels in both trials, and CD4+, CD8+ T cells subsets and interleukin-1beta production were tested in Exp. II. In Exp. I both average daily gain and average daily feed intake of weaned pigs were improved quadratically and feed efficiency was increased linearly by CLA supplementation. Lymphocyte proliferation response to concanavalin A was increased quadratically as dietary CLA concentration increased on day 14 and 28. Ovalbumin antibody production levels were increased linearly on day 7 after injection of ovalbumin and increased quadratically on day 14 after injection, which follows the increased CLA levels, whereas CLA reduced linearly the production of prostaglandin E2 (PGE2). The results of Exp. II indicated that CLA improved performance, lymphocyte proliferation, and increased the CD8+ lymphocyte population, while reduced the production of PGE2 and interleukin-lbeta (IL- 1beta). These results suggest that the supplementation of CLA enhanced lymphocyte proliferation function, possibly by regulating the PGE2 production, and improved growth performance of pigs. Further studies are needed to determine the mechanism of CLA induced inhibition of IL-1beta production. PMID- 15889652 TI - Phosphorus equivalence of a Consensus phytase produced by Hansenula polymorpha in diets for young turkeys. AB - An experiment was conducted to determine the efficacy of a so-called consensus phytase preparation produced by Hansenula polymorpha on growth, tibia and toe ash and P retention of young turkeys. A total of 192 female turkeys (BUT 9 strain) were placed into 96 batteries at two per cage and assigned to one of eight diets: A negative control containing 2.5 g non-phytate P per kg feed (T-1); T-1 plus 125, 250, 500, 1000 or 10 000 phytase units (U) per kg feed, respectively, (T-2 to T-6); T-1 plus 0.5 or 1.0 g Pi/kg feed as dicalcium phosphate dihydrate (DCP), respectively, (T-7 and 8). The experiment lasted 32 d, and excreta were collected and weighed between 26 and 29 d of age. Feed was also weighed during this period in order to calculate P retention. Performance was calculated for the 0 to 32 d period. At the end of the experiment one bird per pen was killed for tibia and toe ash content determination, except for T-2, T-3, and T-7. Body weight, FCR, toe and tibia ash, and P retention responded to phytase or Pi supplementation. Using linear or quadratic models for comparing performance of the treatments containing supplemental Pi with phytase treatments, an equivalence between phytase and Pi was calculated. Body weight, toe ash, tibia ash and P retention showed a significant response to phytase supplementation. The values of equivalence for body weight, toe ash, tibia ash and P retention were 251, 597, 391 and 390 U to 1 g Pi/kg feed. At 10 000 U/kg feed there was a significant response in terms of weight gain and P retention, indicating that turkeys respond to levels greater than 1 000 U/kg feed. PMID- 15889653 TI - Ileal and total tract nutrient digestibility in blue foxes (Alopex lagopus) fed extruded diets containing different protein sources. AB - This study compared apparent ileal and total tract digestibility of macronutrients and amino acids in blue foxes (Alopex lagopus) fed dry extruded diets. The control diet contained fish meal as the main protein source, while in the other diets 50% of the fish meal protein was replaced by not de-hulled solvent-extracted soybean meal, meat meal or bacterial protein meal. Ileal digestibility was obtained with animals modified with ileorectal anastomosis (IRA method). There were no significant differences in the average digestibility of CP, Lys, Trp, Val, Ala or Glu between IRA modified and intact animals. Average ileal digestibilities of His, Thr, Asp, Cys, Gly, Hyp, Pro, Ser, starch and total carbohydrates (CHO) were significantly lower compared with total tract values. Average ileal digestibility of Arg, Ile, Leu, Met, Phe, Tyr and crude fat was significantly higher than total tract digestibility. Digestibility measured in IRA modified animals showed less variability compared with values from intact animals. There were significant differences among diets in ileal as well as total tract digestibility of orude protein, most amino acids, starch and CHO, and in ileal digestibility of crude fat. Ileal crude protein digestibility of the diets ranged from 81.0-86.4% and total tract digestibility from 82.5-86.4%. Ileal amino acid digestibilities ranged from 70.1 (Asp) to 93.3% (Arg) and total tract values ranged from 72.8 (Cys) to 92.2% (Arg). Both ileal and total tract digestibilities of crude protein and most amino acids were significantly lower for diets containing meat meal or bacterial protein meal compared with the control fish meal-based diet and the diet with soybean meal. Although ileal digestibility may be more accurate than total tract digestibility in estimating the protein and amino acid supply in blue foxes, total tract digestibility may be acceptable because of numerically small differences between ileal and total tract digestibilities for protein and most amino acids. It was further concluded that ileal digestibilities of starch and CHO in the blue fox are lower than corresponding total tract digestibilities. PMID- 15889654 TI - Effect of glyphosate contaminated feed on rumen fermentation parameters and in sacco degradation of grass hay and corn grain. AB - Abstract Four rumen fistulated wethers were used to investigate the effect of glyphosate contaminated feed on rumen fermentation. The rations were based on corn silage, urea and a vitamin-mineral premix, either in the absence or presence of 0.77 g glyphosate per kg DM. Furthermore, rations were fed either with or without aromatic amino acid supplementation. During four periods of 28 days, sheep received each of the four dietary treatments according to a Latin square. After 14 days of adaptation rumen fermentation parameters (pH, ammonia, volatile fatty acids) were measured on day 15 over a five-hour period after the morning feeding. The remaining 13 days served for in sacco degradation studies with grass hay and corn grain. Ammonia (NH3) and pH of rumen fluid were within the normal range for all dietary treatments (NH3: 9.1-32.3 mmol x l(- l), pH: 6.2-6.7). Neither rumen fermentation parameters nor in sacco DM and NDF degradation of incubated feedstuffs were significantly affected by glyphosate, with or without aromatic amino acid supplementation. Kinetic profiles of the in sacco dry matter and NDF degradation of grass hay were almost identical for the dietary treatments. PMID- 15889655 TI - On enduring visions: Peter Scharer's and the international college of prosthodontists. PMID- 15889656 TI - Patient satisfaction and preference with magnet, bar-clip, and ball-socket retained mandibular implant overdentures: a cross-over clinical trial. AB - PURPOSE: The purposes of this study were to: (1) determine patient satisfaction with implant-supported mandibular overdentures using magnet, bar-clip, and ball socket attachments; and (2) assess the relation between maximum bite force and patient satisfaction. MATERIALS AND METHODS: In a cross-over clinical trial, 18 edentulous patients with mandibular denture complaints received two mandibular implants and new mandibular and maxillary dentures. The mandibular denture was initially without any kind of attachment system, but it was fitted with one of the attachment types after 3 months. The attachments were changed 3 months thereafter, in random order. A questionnaire on denture complaints was administered at baseline (with the old denture), after 3 months of function with the new denture without attachments, and after 3 months of function with each of the attachments (within-subject comparison). In addition, patients were asked to express their overall appreciation of their dentures on a VAS. Patients' preferences were determined at the end of the experiment. Maximum bite forces were obtained from a previous study with the same population. Five scales of denture complaints were constructed. Mean scale and VAS scores at the five evaluation points were compared among the groups. Pearson correlation was calculated between maximum bite force and scale and VAS scores. RESULTS AND CONCLUSION: Mandibular implant-supported overdenture treatment reduced various denture complaints. The VAS score better reflected patients' preferences than did scale score. Patients strongly preferred bar-clip (10/18 subjects) and ball socket attachments (7/18 subjects) over magnet attachments (1/18 subjects). Patients' preferences could not be predicted on the basis of baseline observations. Maximum bite force was not correlated to scale or VAS score. Hence, patients with higher maximum bite forces were not necessarily more satisfied. PMID- 15889657 TI - Oral health impact in patients wearing removable prostheses: relations to somatization, pain sensitivity, and body consciousness. AB - PURPOSE: Previous studies investigating associations between patient personality traits and complaints related to wearing dental prostheses have been inconclusive. From the perspective of cognitive behavioral theory, the current study investigated whether pain sensitivity, body consciousness, and somatization affected the oral health of patients wearing removable dentures. MATERIALS AND METHODS: Eighty-eight patients were supplied with removable partial and complete dentures. The Oral Health Impact Profile (OHIP), with six subscales measuring oral health impairment and disability during daily living, the Pain Sensitivity Index, the Private Body Consciousness scale, and the Somatization Scale of the SCL-90-R, were used. RESULTS: The variables pain sensitivity, body consciousness, and somatization correlated significantly with all six OHIP subscales in removable denture wearers. In multiple hierarchic regression analyses, patient personality accounted for 38.0% of functional limitation and 41.5% of physical pain. CONCLUSION: Pain sensitivity and bodily preoccupation might be important factors in explaining the subjective oral health effects of removable denture wearing. PMID- 15889658 TI - Influence of self-etching primer preapplication on the dentin-titanium shear bond strength mediated by a resin-modified glass-lonomer cement. AB - PURPOSE: The aims of this study were to evaluate the in vitro shear bond strength of the dentin-titanium interface mediated by Fuji Plus, a resin-modified glass ionomer cement, applied in combination with several self-etching bonding systems, and to compare the values obtained with those of SuperBond, chosen as a reference luting agent. MATERIALS AND METHODS: The self-etching bonding systems used in combination with Fuji Plus were AdheSE, Xeno III, Clearfil SEBond, Unifil Bond, and an experimental self-etching bonding system comprising the sequential application of Xeno III and the bonding component of SE Bond. Two control groups were also tested: a negative control with Fuji Plus along with its conditioner (10% ferric chloride, 2% citric acid solution); and a positive control with the resin cement SuperBond. One hundred five human molars were used to prepare seven groups of fifteen samples each. The samples were tested in shear after 7-day storage at 100% relative humidity and 37 degrees C. RESULTS AND CONCLUSION: Three of the tested combinations of self-etching bonding systems with Fuji Plus significantly improved in comparison with the negative control group, from 9% to 44%. The self-etching bonding systems obtained using Fuji Plus in combination with Clearfil SEBond, Unifil Bond, and the experimental system were not different from those obtained with SuperBond. PMID- 15889659 TI - Long-term treatment costs associated with implant-supported mandibular prostheses in edentulous patients. AB - PURPOSE: The study's aim was to report long-term costs in edentulous patients treated with mandibular implant-supported prostheses. MATERIALS AND METHODS: Ninety patients were divided into four groups based on the type of implant prosthesis (fixed or overdenture) and treatment year. Records were obtained from dental charts, and an economic analysis from the patient's perspective was conducted. Clinical time associated with various procedures was measured and applied to the four groups. Salary rates by age, occupation, and gender were used to value patients' time. Direct clinical and time costs over 10 years were converted to 2002 Canadian dollars using the Consumer Price Index and discounted at a 3% rate. A sensitivity analysis at an equal salary rate was carried out to test the robustness of the time costs. RESULTS: Initial treatment and maintenance costs over the observation period were significantly higher for fixed compared to overdenture prostheses. A significant improvement in maintenance costs for the first patient group treated with fixed prostheses was observed over the follow-up period. Longer term (15 years) treatment costs for the initial two groups were significantly higher for the fixed group. The sensitivity analysis at an equal salary rate demonstrated the same trend: Time costs were significantly higher for the fixed groups. CONCLUSION: Long-term treatment costs indicated that the mandibular overdenture was a less expensive treatment compared to the fixed implant prosthesis. PMID- 15889660 TI - Problems reported by patients before and after prosthodontic treatment. AB - PURPOSE: This study sought to investigate problems reported by patients before and after prosthodontic treatment. MATERIALS AND METHODS: Patient-reported problems were studied using the item list contained in the German version of the Oral Health Impact Profile in a convenience sample of 107 prosthodontic patients before (T0), 1 month after (T1), and 6 to 12 months after treatment (T2). "Frequently reported" problems were defined as impacts experienced fairly often or very often. The prevalence of frequently reported problems was compared among treatment groups and across appointments. RESULTS: At baseline, the most prevalent frequently reported problems were "difficulty chewing" (31%), "take longer to complete a meal" (28%), "food catching" (26%), "uncomfortable to eat" (24%), and "unable to eat (because of dentures)" (23%). At T2, the most prevalent frequently reported problems were "sore spots" (5%), "painful gums" (4%), "discomfort (because of dentures)" (3%), and "sore jaw" (2%). The number of reported problems decreased from 18.0 (T0) to 7.5 (T1), and further to 4.5 (T2). The decrease was the fastest in fixed partial denture wearers and the slowest in removable partial denture wearers. Some problems emerged during or after prosthodontic treatment. The kind of pre- and posttreatment problems differed substantially. CONCLUSION: The number of problems decreased substantially after prosthodontic treatment. Fixed partial dentures more effectively influenced the problems reported before treatment than did removable partial or complete dentures. PMID- 15889661 TI - Early experience of implant-supported prostheses in patients with neurologic disabilities. AB - PURPOSE: The purpose of this prospective study was to provide a preliminary report of implant treatment in patients with neurologic disabilities. MATERIALS AND METHODS: Patients had been referred to the National Orofacial Resource Centre with different disabilities, specifically neurologic disorders causing various orofacial dysfunction problems, and were suitable for prosthodontic treatment with implants. Patients were treated with single implant-supported crowns, fixed partial dentures, or complete implant-supported dentures. Implants used were threaded titanium cylinders placed under general or local anesthesia. All surgical complications in the healing period or at second-stage surgery were noted, as were all prosthodontic complications. RESULTS: Fourteen patients were treated with 35 implants. Three implants were lost before or at second-stage surgery, and two implants were lost after loading. These complications were observed in two patients. Implant failures and other complications were observed in two patients in the mandible because of dehiscence of the mucoperiosteal flap and infection. Some patients had developed finger or oral habits, such as excessive tongue movements, that were probably responsible for these complications. One of the patients with Down syndrome possibly had reduced resistance to infections. No major complications were observed for the other 12 patients treated. CONCLUSION: Strict adherence to a surgical protocol is needed for the management of patients with neurologic disabilities. It is important to inform the patient's caregiver about maintenance of good oral hygiene and the increased risk of complications caused by finger or oral habits. PMID- 15889662 TI - Indications for removable partial dentures: a literature review. AB - PURPOSE: The purpose of this report was to evaluate indications for the use of removable partial dentures based on a comprehensive literature review. MATERIALS AND METHODS: Using a model similar to a Delphi process, the literature relating to the indications and contraindications for the prescription of removable partial dentures was discussed by seven experienced educators in a 2.5-day workshop. RESULTS: Evidence for indications and contraindications for the prescription of removable partial dentures is not clearly stated in the literature; however, some basic principles are defined. There appears to be a trend in favor of the use of the shortened dental arch concept or implant supported restorations instead of conventional removable partial dentures, given the evidence that the long-term use of removable dentures is associated with increased risks of caries and periodontitis and low patient acceptance. The presence of sound abutment teeth appears to encourage the use of removable partial dentures, as the fixed partial denture alternative requires sacrifice of healthy hard tissues. When economic factors influence the decision-making process, removable partial dentures are often chosen. CONCLUSION: While evidence based indications and contraindications for prescribing removable partial dentures are still lacking, major underlying principles for clinical decision making are identified. PMID- 15889663 TI - Influence of mental status on removable prosthesis compliance in institutionalized elderly persons. AB - PURPOSE: It is well-recognized that many institutionalized elderly persons with dementia do not wear dentures. The objective of this study was to evaluate the current status of denture use among elderly patients with dementia, in association with degree of mental impairment, to provide information relating to a decision-making process for optimal denture treatment. MATERIALS AND METHODS: From August 2001 to December 2002, 101 hospitalized elderly persons in a geriatric hospital dental clinic received removable denture treatment by two certified prosthodontists. Prior to denture treatment, patient mental status was evaluated using the Mini Mental Status Examination (MMSE). Basic activities of daily living, including mobility, feeding, toilet use, dressing, and bathing, were also evaluated. Denture acceptance was determined 6 months after denture delivery. RESULTS: Eight patients were excluded; 73 patients had accepted their dentures and 20 had not 6 months after denture delivery. The mean MMSE score for patients who did not accept denture delivery (11.7+/-7.0) was significantly lower than that of those who did accept and wear their dentures (16.0+/-6.8). CONCLUSION: The cognitive status of institutionalized elderly persons with dementia should be a criterion for clinical decision making relating to denture treatment. PMID- 15889664 TI - Comparison of CIE lab, CIEDE 2000, and DIN 99 color differences between various shades of resin composites. AB - PURPOSE: In addition to the CIE Lab color-difference formula (deltaE*ab), advanced formulas that include weighting functions have been introduced. The objectives of this study were to determine the correlations in color differences between different shade pairs of resin composites by different color formulas, and to determine whether the weighting functions included in the advanced formulas influenced the color-difference values. MATERIALS AND METHODS: Color was measured after polymerization of two resin composites in 26 shades, and color differences between shades were calculated by deltaE*ab, CIEDE 2000 (deltaE00), and DIN 99 (deltaE99) formulas. Regression analyses were performed between the color differences calculated by three formulas in each group divided by the differences in color parametric factors between the pairs compared. RESULTS: There were significant correlations between deltaEab and deltaE00, deltaEab and deltaE99, and deltaE99 and deltaE00 (r2 = .99, .89, and .90, respectively). The weighting functions in the CIEDE 2000 formula had influence on deltaE00 values when differences in chroma and hue between compared pairs were great. CONCLUSION: Differences in parametric factors between the pairs compared influenced the correlation between deltaE*ab and deltaE99 values (r2 = .25 to .97). As the CIEDE 2000 formula has been proven to be better matched to observer responses and showed significant involvement of weighting functions compared to CIE Lab color difference, this formula should be considered for evaluation of the color of resin composites. PMID- 15889665 TI - Evaluation of 24-hour shear bond strength of resin composite to porcelain according to surface treatment. AB - PURPOSE: The purpose of the present in vitro study was to examine shear bond strengths of resin composite to porcelain according to surface treatment. MATERIALS AND METHODS: One hundred eight feldspathic porcelain blocks were prepared. Specimens were divided into nine surface treatment groups: sandblasting with 50- or 110-microm Al2O3, acid etching with hydrofluoric acid, applying silane agent, and combinations of these treatments. Microhybrid resin composite was condensed and light cured for 40 seconds on the porcelain specimens, which were then stored in distilled water at 37 degrees C for 24 hours before mechanical testing. RESULTS: The bond strengths were significantly different according to one-way ANOVA. The lowest bond strength was observed in the silane group (4.09 MPa); the highest bond strengths were observed with acid etching + silane (11.97 MPa) and sandblasting with 50-microm Al2O3 + acid etching + silane (12.34 MPa) (no significant difference between groups). CONCLUSION: Acid etching with 9.6% hydrofluoric acid gel or sandblasting with 50- or 110-microm Al2O3 particles alone did not provide adequate bond strength. Silane agent was effective in increasing the shear bond strength of resin composite to porcelain after sandblasting or acid etching. Porcelain treatment with a combination of 50 microm Al2O3 air abrasion, 9.6% hydrofluoric acid, and silane agent provided higher bond strengths than treatment with any of these procedures alone. PMID- 15889666 TI - Fracture risk judgment and crown indication by teachers in a dental school: a pilot study. PMID- 15889667 TI - Prosthodontic and endodontic management of a patient with skeletal class III malocclusion: a clinical report. PMID- 15889668 TI - Comparison of external and internal hex implants' rotational freedom: a pilot study. PMID- 15889669 TI - [Hormone resistance and its modulation in breast cancer]. AB - Acquired hormone resistance is a reversible adaptative change of hormone sensitive breast tumors promoting survival by changes in the balance and communication between estrogen receptor and growth factor signaling. The mechanisms of hormone resistance induced by various hormone therapies are different, however, their common feature is the dominance of growth factor signaling with the consequences of enhanced proliferation and decreased apoptosis. In case of tamoxifen or selective estrogen receptor modulator resistance, the agents' enhanced agonistic activity occurs. The increased expression of certain estrogen receptor coactivators may play an important role. The essential of hormone resistance after estrogen deprivation is estrogen hypersensitivity, which is a consequence of the enhanced activity of the membrane associated estrogen receptor and its influence on the growth factor signaling. The integration of cell surface growth factor receptor or growth factor signal transduction blocking agents like tyrosine kinase, MAPK, mTOR, PI3K or farnesyl transferase inhibitors into hormone therapies may prevent or treat hormone resistance. The other possibility is to use the hormone therapies sequentially. A new promising agent is the pure antiestrogen fulvestrant which targets the estrogen receptor located in both the membrane or the nucleus. Also, estrogen therapy may revert hormone resistance. The use of predictive markers may promote treatment choice and indicate application of targeted therapies. PMID- 15889670 TI - [Significance of cytochrome P450 2C9 genotype for the bleeding complications in patients treated with acenocoumarol]. AB - INTRODUCTION: For the primary and secondary prevention of thromboembolic events are used the oral anticoagulants, the drugs having a low therapeutic index and frequent bleeding complication rate. Establishing the proper therapeutic dose of these drugs for different patients is complicated by a variety of conditions, such as the comorbidity, age, other drugs used, diet, and pharmacogenetic factors. One of the latters is the polymorphism of the cytochrome P450 CYP2C9 enzyme. AIM: The influence of CYP2C9 polymorphism on the effectiveness of the--in Hungary for oral anticoagulation exclusively used--acenocoumarol therapy and on the occurrence of bleeding complications was investigated. METHODS: Genotyping of 421 patients including 183 men and 238 women, (mean age 66.2 +/- 11.8 years) who took acenocoumarol (Syncumar) for at least 6 months was performed. Based on anamnestic and laboratory data, the correlation between the genotype and the acenocoumarol dose and bleeding complications were retrospectively analysed. RESULTS: The frequency-distribution for the CYP2C9*1, *2, and *3 alleles were found to be: 0.814, 0.110, and 0.076, respectively. In the 145 patients bearing the alleles with reduced activity (CYP2C9*2 and/or *3), the optimised dose of the acenocoumarol was significantly (p < 0.001) lower than in patients with the wild type allele (2.12 +/- 0.96 mg/day and 2.90 +/- 1.45 mg/day, respectively). Although the occurrence of minor bleeding complications in the former group was significantly (p < 0.005) higher [OR = 1.99 (CI: 1.20-3.33)], there was no difference in major bleeding complications. In patients taking an acenocoumarol dose lower than 2 mg/day, the occurrence of an INR value higher than 6 in the anamnesis was significantly (p < 0.05) more frequent. Evaluating separately the variant alleles we have concluded, that in the presence of allele *2 a lower acenocoumarol dose was required than in wild-type subjects, and even lower in the presence of allele *3. CONCLUSIONS: The frequency-distribution of the CYP2C9 alleles was as reported by others. In patients bearing alleles with reduced enzymatic activity, the occurrence of minor bleeding complications and the INR values higher than 6 were significantly more frequent. In patients with a lower acenocoumarol demand at the introduction of this therapy, a caution is required. In order to test the hypothesis that before the initiation of acenocoumarol therapy the determination of CYP2C9 polymorphism is cost-effective and could improve the optimization of anticoagulation and reduce the risk of bleeding complications a large prospective randomised trial is required. PMID- 15889671 TI - [Left ventricular systolic and diastolic function and dysfunction]. AB - The left ventricular systolic function--independently from the type of cardiac disease--determines the quality of life and the prognosis of the patients, so the examination of this parameter is indispensable. To measure it accurately is difficult, because the muscle fibers in the ventricular wall run partly circumferentially, partly longitudinally and tangentially and we have not any kind of method, which could have examined the function all of these differently contracting myocardial muscle fibers. To add these, the shape of the left ventricle is geometrically irregular, all ventricular segments move toward a "gravitational point" during the systole and backward during the diastole, and the heart also has a rotational motion in the chest during every cardiac cycle. Haemodynamic parameters (preload, afterload, heart rate and synchronous or asynchronous motion of the left ventricular walls) also can modify the left ventricular function. The ejection fraction is most frequently measured by echocardiography. M-mode, 2-dimensional, 3-dimensional echocardiography, Doppler methods, Doppler Myocardial Imaging (DMI), contrast echocardiography and color kinesis can be used to get more precise information about the left ventricular systolic function. The diastolic left ventricular function can be analysed using the mitral Doppler curve. The DMI, Strain Rate Imaging (SRI), contrast echocardiography and color kinesis improve the quality of imaging and so the accuracy of measurement. Using Doppler Myocardial Imaging and Strain Rate Imaging we can get information about the global and segmental left ventricular systolic and diastolic function. These parameters are well reproducible, and that is why they can be used in the assessment of the efficacy of pharmaceutical or other therapeutical procedures and following the changes in the clinical state of the patients. PMID- 15889672 TI - [Posterior cloaca with accessory urethra]. AB - The authors present a case of posterior cloaca in an 8-year-old girl. The diagnosis of this rare anomaly requires meticulous examination of the female genitalia and detailed radiological investigation. She underwent total urogenital mobilization (TUM) without opening the anal canal and the rectum. Eighteen months of follow up period she continues to have full bowel and urinary control with almost normal looking genitalia. PMID- 15889673 TI - [Augustine Schopf-Merei, medical historian]. PMID- 15889674 TI - [Contribution to the past of health care in Karpatalja (I.)]. PMID- 15889675 TI - [Dezso Velits, biographical data and appreciation of his work]. PMID- 15889676 TI - [Contribution to the article "Human alveolar echinococcosis"]. PMID- 15889677 TI - [Participation in the European Union Research Program. Aladdin is not from a fairy tale]. PMID- 15889678 TI - Proxy consent in neonatal care--goal-directed or procedure-specific? AB - The prescription of practice guidelines for consent in neonatal care that are appropriate for all interventions faces substantial problems. Current practice varies widely. Consent in neonatal care is compromised by postnatal constraints on information sharing and decision-making. Empirical research shows marked individual and cultural variation in the degree to which parents want to contribute to decision-making on behalf of their infants. Conflict between the parents' wishes and the infant's best interests could arise if consent for a recommended intervention were refused, and parental refusal of consent may have to be overridden. Consent to an appropriate package of care (such as special, intensive or palliative care) may be morally preferable to a universal requirement to seek consent for all individual interventions entailed by that package. PMID- 15889679 TI - Commercial agencies and surrogate motherhood: a transaction cost approach. AB - In this paper we investigate the legal arrangements involved in UK surrogate motherhood from a transaction-cost perspective. We outline the specific forms the transaction costs take and critically comment on the way in which the UK institutional and organisational arrangements at present adversely influence transaction costs. We then focus specifically on the potential role of surrogacy agencies and look at UK and US evidence on commercial and voluntary agencies. Policy implications follow. PMID- 15889680 TI - Clarifying efficiency-equity tradeoffs through explicit criteria, with a focus on developing countries. AB - Expenditures on health in many developing countries are being disproportionately spent on health services that have a low overall health impact, and that disproportionately benefit the rich. Without explicit consideration of priority setting, this situation is likely to remain unchanged: resource allocation is too often dictated by historical patterns, and maintains vested interests. This paper explores how prioritization between different health interventions can be rationalised by the use of clearly defined criteria. A number of key efficiency and equity criteria are examined, in particular analysing how potential tradeoffs could be incorporated into the decision making process. PMID- 15889681 TI - Priorities in the use of research into ageing. AB - This paper considers which applications of research into ageing should be supported. It assumes that both applications which enhance the quality of life for the elderly and applications which extend the life-span are desirable, and then considers which should be prioritised. It is argued that in the present state of our knowledge and under present social and medical conditions there are a number of reasons for favouring the improvement of the quality of life over increasing the life-span, and thinking that this is likely to do more good and for more people. PMID- 15889682 TI - Evidence-based medicine: why do opponents and proponents use the same arguments? AB - There is quite some ethical controversy on Evidence-based Medicine (EbM) with regard to issues of physician autonomy as well as its allocative implications. Yet, there are some shortcomings in the current debate. First of all, some of the arguments brought up against EbM are similarly defaults of "classical medicine" as well, for instance its negligence of social aspects of medicine. Second, it is often maintained that EbM is just a tool to attain cost containment. This argument is false in two regards for neither is there any idea of cutting costs in the roots of EbM nor does EbM once practiced necessarily lead to less costs as there can be underuse as well as overuse. Third, both opponents and proponents of EbM come up with the same arguments against each other. Both maintain that the other way of practicing medicine does not allow for physicians' autonomy and free judgment. Therefore, we are going to search for the different presuppositions on which these "reproaches" rely. In this way we can demonstrate that both opponents and proponents rely on different notions of autonomy and free judgment in their argument. Finally, we hope to show that some of the ethical criticism may be raised against classical medicine as well and that allocation in terms of costs is not primarily an aim of EbM. PMID- 15889683 TI - Use and abuse of bioethics: integrity and professional standing. AB - This paper sets out to examine the integrity and professional standing of "Bioethics." It argues that professions have certain responsibilities that start with setting criteria for and credentialing those that have met the criteria and goes on to ultimately have social responsibilities to the community. As it now stands we claim that Bioethics--while it certainly has achieved some progress in the way medicine has developed--has failed to become a profession and has to a large extent failed in its social responsibility. We feel that Bioethics has to define itself, set criteria for membership in the profession, police itself and- above all--meet its social responsibility to become a profession meriting that name. PMID- 15889684 TI - Sex selection: laissez faire or family balancing? AB - In a recent comment on the HFEA's public consultation on sex selection, Soren Holm claimed that proponents of family balancing are committed to embrace a laissez faire approach. Given that arguments in support of sex selection for family balancing also support sex selection for other social reasons, advocates of family balancing, he asserts, are simply inconsistent when calling for a limit on access to sex selection. In this paper, I argue that proponents of family balancing are in no way inconsistent. Provided their advocacy of family balancing is aimed at preventing a severe distortion of the natural sex ratio, they are entirely justified in insisting on restrictions to sex selection. The real question is whether a concern for the sex ratio does indeed call for a limit on sex selection. Based on a recent survey on gender preferences and data from several Gender Clinics, I argue that a restriction on sex selection to family balancing is unwarranted. In the absence of any evidence for a pending sex ratio distortion, we are actually required to adopt a laissez faire approach to sex selection. PMID- 15889685 TI - Patients reading about themselves: a stimulus to psychoanalytic work. AB - Patients' reading of their psychoanalysts' papers about them as a vehicle for psychoanalytic work is a relatively new phenomenon in the field. Over the past five years, reports of analysts' employment of their writing in this fashion have begun to appear in the analytic literature. This paper presents clinical illustrations of this specific use of analysts' writing. These illustrations were drawn from interviews with analysts who published clinical articles in Psychoanalytic Dialogues between 1995 and 2003. The author considers some of the clinical and scientific implications of this use of papers written for publication. PMID- 15889686 TI - The cognitive effects of trauma: reversal of alpha function and the formation of a beta screen. AB - Following a brief review of Freud's writings on trauma, the author discusses relevant theories of Bion, and in particular the concepts of the alpha function and the beta screen. A clinical example is presented in which the patient's relatively recent trauma in adulthood had become fused with prior related experiences, leading to a propensity for repeated enactments in analysis and a failure to learn from experience. Drawing on the analyst's alpha function, the patient was gradually able to use mentalization to transform her rigidly structured traumatic organization. The author highlights the roles of dreams/dream associations and of screen memories in the patient's analysis. PMID- 15889687 TI - Which reality in the psychoanalytic session? AB - The author discusses how the analyst's theory influences what happens in the psychoanalytic session, and, in particular, how it determines which of several possible realities become the focus of attention. An overview of Bionian thought and how it informs the author's way of working is provided. In elaborating his use of the narrative level in the session and other key concepts, the author presents three brief clinical vignettes. PMID- 15889688 TI - Clinical conversations between psychoanalysis and imaginative literature. AB - A literary form created by an imaginative writer captures something of the way the author shapes emotional experience and psychologically engages with it. The manner in which experience is created and contained in an imaginative literary text has much in common with the way experience is generated and worked with in the psychoanalytic situation. The author describes a clinical experience in which there was a collapse of the analytic (imaginative) space. He then discusses how he made use of a "conversation" he created with a short story and his own analytic experience to restore his imaginative capacities and to resume psychological work with the patient. PMID- 15889689 TI - A parapraxis in a dream. AB - When a parapraxis is put on display in a dream, one can only wonder what service the willful mistake is rendering to resourceful dream work. Freud taught us that anything that appears in the manifest content of a dream may well be a disguise or a distortion of a subject that originally made an anxiety-provoking, and hence short-lived, first appearance in latent dream thoughts. Dreams in dreams and jokes in dreams have been examined from this perspective (Mahon 2002a, 2002b), and this paper focuses on the appearance and meaning of a parapraxis in a dream, with the argument that seemingly casual "mistakes" are highlighted in the manifest display to cover up some latent, much more deliberate subject matter. PMID- 15889690 TI - Sanderian activation waves: a hypothesis of a nonsymbolic influence on moods. AB - This paper describes a hypothesis of Tronick's (2002) that positive and negative affect activation waves with changing amplitudes based on positive and negative lived experiences are the sources of persistent or regularly recurrent moods of feeling upbeat or downbeat. The characteristics of discrete affects, moods, and affect states are discussed in the light of a previous study based on clinical observation (Lichtenberg, Lachmann, and Fosshage 1996). The clinical experience theory underpinning that study is then compared with Tronick's Sanderian activation wave theory. A case example is presented, and the overall relevance of Tronick's theory to clinical work is discussed. PMID- 15889691 TI - On putting a cloud in a bottle: psychoanalytic perspectives on mysticism. AB - This essay addresses the psychoanalytic understanding of mystical phenomena and mystical ecstatic states. The study of mystical phenomena is a core area of interest in the ongoing dialogue between psychoanalysis and religion. Mystical phenomena and states are discussed in this essay, with particular reference to descriptions found in an anonymous medieval tract, The Cloud of Unknowing. Some common psychoanalytic views of mystical phenomena are discussed and criticized, and an effort is made to formulate an approach to the understanding of these extraordinary mental and spiritual states that is both more congruent with accounts of authentic mystics and more consistent with psychoanalytic principles. PMID- 15889692 TI - In a bid to restate the culture-psyche problematic: revisiting The Essential Writings of Suhir Kakar. AB - This book assembles a broad sweep of its author's writings in one volume. Sudhir Kakar is an erudite psychoanalyst who has written nearly fifteen previous books on various aspects of Indian psyche and culture. Most are concerned with alternative healing practices and traditions, aspects of Indian childhood, and the place of sexuality in Indian culture. Informed by psychohistory and influenced by Erikson's theory of identity and its psychosocial development within the life cycle of an individual in a given society, Kakar accurately articulates issues of identity and separation-individuation dynamics that operate within Indian psyche and culture. In discussing the subject book, the author of this essay applies a psychoanalytically focused approach, emphasizing the teachings of French psychoanalysis and the British object relations school in particular. PMID- 15889693 TI - Rewarding quality care: the advent of performance-based payment initiatives. PMID- 15889694 TI - Reproductive issues for women with spina bifida. AB - BACKGROUND/OBJECTIVES: Few studies have adequately examined the unique issues of women with spina bifida as they enter their reproductive years. Most studies are anecdotal, retrospective case studies that contribute little to our understanding of the physiological effects of the disability on the reproductive system and, conversely, the effects of the reproductive endocrine changes on the woman's disability. The purpose of this article was to review previously published reports on the reproductive issues facing female adolescents and women and to ascertain the current knowledge so that future research needs can be established. METHODS: Multiple MEDLINE searches were performed for publications from 1957 through the present pertaining to females/women with spina bifida and/or disabilities. Papers were then chosen if information was provided on menarche, menstruation and sexual maturation, gynecologic issues, sexual function, birth control, pregnancy outcomes, and menopause. These were then reviewed and classified according to the level of evidence (as defined by Ball et al). RESULTS: More than 150 titles and abstracts were reviewed for inclusion. Of these, 75 papers met the guidelines for the subject content. Studies were stratified by level of evidence and included 1 study at level 3, 71 studies at level 4, and 3 studies at level 5. CONCLUSION: Little research has examined the reproductive issues of women with spina bifida and women with other neurologic disabilities. Future controlled prospective research studies are needed to examine issues related to puberty and sexual and gynecologic issues throughout the life span and pregnancy in these populations. PMID- 15889695 TI - Epidemiology and demography of acute spinal cord injury in a large urban setting. AB - OBJECTIVE: In a large, single-center study of subjects with acute traumatic spinal cord injury (SCI), we describe the sample population with respect to gender, age, cause of injury, and severity of injury, to see whether these properties are similar to those of other large-scale studies of acute SCI conducted in the past. METHODS: As part of a study to examine the natural pattern of recovery after acute SCI (presented elsewhere), descriptive information was gathered in relation to subject population and injury properties. RESULTS: A total of 229 subjects were recruited. The study population had a higher percentage of women and a higher mean age of men and women than those of most other published studies of acute SCI. A greater percentage of incomplete subjects was also encountered. The incidence of gunshot injury as a cause of SCI was considerably lower in this study than had been the case 10 years previously in Dade County, Florida. CONCLUSIONS: The demography of acute SCI within a major urban center of South Florida suggests a trend toward less severe injury than in years past. These findings support the development of animal models for testing SCI treatment that include cohorts having mild to moderate injury severity, in order to achieve greater clinical relevance. PMID- 15889696 TI - Appearance of a novel prostacyclin receptor antibody and duration of spinal cord injury. AB - BACKGROUND: Cardiovascular disease (CVD) appears to be accelerated in individuals with chronic spinal cord injury (SCI). Previously, we have identified a novel circulating antibody (IgG) in persons with SCI that specifically blocks the high affinity prostacyclin (PGI2) receptors on the platelet surface without affecting the low-affinity PGI2 receptors. OBJECTIVE: In this study, the relationship between the time course after SCI to the development of IgG to the high-affinity PGI2 receptor was determined. METHODS: Blood samples were collected 1, 3, 5, 10, and > 10 (15 +/- 4) years after SCI (n = 36). Plasma samples (50 microg) were analyzed by polyacrylamide gel electrophoresis (PAGE) followed by densitometry. RESULTS: The optical density (OD) of the IgG (molecular weight 47,000) at 1 year after SCI was significantly higher than control (1.65 +/- 0.08 vs 1.33 +/- 0.04; P < 0.01). This anti-receptor IgG appears to increase for 5 years and then plateau. At 5 years, 6-10 years, and > 10 years of injury, the OD was 1.83 +/- 0.09, 1.83 +/- 0.10, and 1.87 +/- 0.08, respectively. With an increase in this specific IgG, there was a concomitant decrease in the binding of prostacyclin to its high-affinity receptors on SCI platelets, (non-SCI vs 1, 3, and 5 years after injury; n1 = 172 +/- 25 vs 153 +/- 15, 107 +/- 25, and 40 +/- 4 sites/platelet, respectively; P < 0.001), with no significant change in receptor affinity. CONCLUSIONS: The level of the high-affinity PGI2-receptor antibody determined in individuals with SCI was associated with the duration and not with the level of injury. Platelets from subjects with SCI had a reduction in numbers of high affinity receptors. PMID- 15889697 TI - Partial angiotensin-converting enzyme inhibition during acute orthostatic stress in persons with tetraplegia. AB - INTRODUCTION: Individuals with tetraplegia rely on the renin-angiotensin system for orthostatic blood pressure control. OBJECTIVES: To determine the effect of partial angiotensin-converting enzyme (ACE) inhibition on heart rate (HR), active plasma renin (PR), and mean arterial blood pressure (MAP) during acute orthostasis in subjects with tetraplegia (n = 7) and nondisabled persons (n = 8). METHODS: Subjects were instructed to avoid caffeine and alcohol for 24 hours before testing and to report to the laboratory between 10 AM and 1 PM. Progressive head-up tilt (15 degrees, 25 degrees, 35 degrees, and 45 degrees) was performed on 2 separate days; Day 1: without ACE inhibition; Day 2: after intravenous (IV) infusion of enalaprilat (0.625 mg). RESULTS: HR was reduced during orthostasis in the tetraplegia compared with the nondisabled group (P < 0.0001), and was unaffected by ACE inhibition in either group. PR was not increased with orthostasis in either group, but was increased after ACE inhibition in both groups (P < 0.001). MAP was not affected by orthostasis in either group, but was reduced with ACE inhibition in both groups (P < 0.01). In the tetraplegia group, MAP was initially reduced after ACE inhibition, but was maintained thereafter with increasing angles of tilt, and no subject complained of symptomatic orthostatic hypotension. CONCLUSION: Subjects with tetraplegia were tolerant of an acute bout of orthostatic stress after partial ACE inhibition. This may have clinical relevance because of the increased prevalence of type 2 diabetes mellitus in this population and the use of ACE inhibitors for the treatment of progressive renal and cardiovascular disease. PMID- 15889698 TI - Reliability of upper extremity anaerobic power assessment in persons with tetraplegia. AB - BACKGROUND/OBJECTIVE: Reliable assessment of upper extremity anaerobic power in persons with cervical spinal cord injury (SCI) may indicate the ability to successfully and safely perform many daily activities. PURPOSE: To examine test retest reliability of upper extremity Wingate anaerobic testing (WAnT) in persons with motor/sensory complete tetraplegia. METHODS: Forty-five persons with cervical-level SCI (15 individuals each at C5, C6, and C7 levels of injury) performed 2-arm WAnT bouts, with 2 to 4 days between bouts. Subjects performed the WAnT seated in their wheelchairs using a tabletop-mounted Monarch 834E ergometer. Resistance loads were applied relative to injury level, with 1%, 2%, and 3% of body mass applied to subjects with C5, C6, and C7 level injuries, respectively. All subjects were directed to crank the ergometer at maximal velocity for a 30-second period. Values of peak power (Ppeak) and mean power (Pmean) were determined using an SMI OptoSensor 2000 system. Ppeak and Pmean were compared between trials and between groups using 2-way analyses of variance for repeated measures. Coefficients of determination (r2) were calculated between trials. RESULTS: There were no significant differences in Ppeak or Pmean detected between the 2 trials of WAnT in the C5, C6, or C7 groups. Regression analyses revealed statistically significant associations between bouts for Pmean and Ppeak in each of the 3 groups (P < 0.05). CONCLUSIONS: Upper extremity WAnT is reliable for upper extremity anaerobic power assessment in persons with cervical SCI at or below the C5 level. PMID- 15889699 TI - Chronic instrumentation with model microstimulators in an animal model of the lower urinary tract. AB - BACKGROUND: Microstimulators are a new type of neuroprosthetic device that should be considered for applications such as micturition control after spinal cord injury (SCI). These devices are small (less than 25 mm by 5 mm) and the electrodes are located on the ends of the stimulator. The aim of the current study was to develop methods for chronic implantation of model microstimulators (M-Micro) on the bladder wall and pelvic plexus of female cats. A postmortem evaluation of the effects of 3 months of implantation is reported. METHODS: Techniques to produce the M-Micro are described. Four of these devices were implanted in 4 female cats and maintained after the initial instrumentation surgery and a second survival surgery for SCI (at T10). Using a single suture tied around the M-Micro, these devices were secured to the bladder wall or the fat pads adjacent to the pelvic plexus. Additional instrumentation was implanted, including 2 catheters in the bladder, 1 abdominal balloon, and electromyography electrodes in the urethral and anal sphincters. Postmortem observations of the location of the M-Micro on the bladder wall were conducted after fixation. RESULTS: The animals' conditions were good. One animal was sacrificed early because of a skin infection. A single suture was sufficient to anchor the M Micro. However, during the surgical implantation the pelvic plexus M-Micro ended up close to the bladder neck. Extensive fibrous connective tissue formed around the M-Micro and implanted catheters on the bladder wall. This appeared to result, in part, from multiple devices implanted on or near the bladder wall. CONCLUSIONS: These pilot studies showed that the M-Micro could be easily constructed and secured to the bladder wall or fat pads close to the pelvic plexus. There was a concern that the pelvic plexus location for the M-Micro ended near the bladder neck during the surgical implantation; however, these devices did not appear to migrate over this short, 3-month implantation period. The extensive connective tissue responses of the bladder wall to the tubes, wires, and M-Micro was a major concern. The M-Micro appears to be a good device to assess the potential of commercial microstimulators for use in micturition control. PMID- 15889700 TI - Fournier gangrene in spinal cord injury: a case report. AB - BACKGROUND: Fournier gangrene is a necrotizing fasciitis of the perineal and genital region resulting from polymicrobial infection in which infection spreads along fascial planes, causing soft-tissue necrosis. If surgical debridement and control of infection are delayed, the disease can progress and result in septic shock, multiorgan failure, and death. Initial symptoms are severe pain in the genital region followed by swelling and erythema. In patients with spinal cord injury (SCI), lack of pain sensation could cause delay in seeking medical attention. SCI patients are at higher risk for Fournier gangrene secondary to neurogenic bladder, neurogenic bowel, and impaired sensation. A literature search resulted in only 1 report of Fournier gangrene with localized necrosis of the scrotum in a patient with SCI. METHODS: Case report of a 47-year-old man with C4 tetraplegia. RESULTS: Patient presented with a necrotic ulceration on the ventral aspect of the penis and scrotum of 2 days duration and was diagnosed with fulminant Fournier gangrene. CONCLUSIONS: Patients with SCI are at higher risk for Fournier gangrene secondary to neurogenic bladder, neurogenic bowel, and impaired sensation. Mortality is high. Prevention and early diagnosis are essential. Prompt aggressive intervention is warranted to maximize outcomes. PMID- 15889701 TI - Overview of spasticity management in multiple sclerosis. Evidence-based management strategies for spasticity treatment in multiple sclerosis. PMID- 15889702 TI - Medical correspondence in early modern Europe. An introduction. PMID- 15889703 TI - The letter: private text or public place? The Mattioli-Gesner controversy about the aconitum primum. AB - From 1555 to 1565, Pietro Andrea Mattioli and Conrad Gesner were locked in controversy over the veracity of Mattioli's picture of aconitum primum. This dispute led to numerous vehement publications and to intensive exchanges of letters, not only between the protagonists but also within their own and sometimes inter-connected networks of correspondence. This dispute illustrates how 16th-century scholars played upon the ambiguous place of these letters between private and public space to deal with controversy in the Republic of Letters. PMID- 15889704 TI - [Postal communications: medicine in the correspondence of Johann Jakob Scheuchzer (1672--1733)]. AB - This article examines different aspects of the history of medicine in the correspondence of the Swiss physician and scientist Johann Jakob Scheuchzer (1672 -1733) from Zurich. The focus is on three different levels: persons, contents and functions. On all three levels we can observe that "medicine" takes in some way part of the res publica litteraria. The international correspondence network of Scheuchzer can be seen as a crucial "multiplicator" of communication within the scientific community. PMID- 15889705 TI - [The professor and his student. The correspondence between Lorenz Heister and Christoph Jacob Trew]. AB - Letters were the central medium of communication in the medical scientific community of the 18th century. Professional as well as personal relationships were established among the various correspondents. These relationships constituted the smallest units of communication which contributed to the regional and international scientific network of the Republic of Letters. A correspondence that grew out of a trusted teacher-student relationship could gain an especially intense character both intellectually and personally. This contribution offers an analysis of an example of just such a correspondence. Lorenz Heister (1683- 1758), medical professor at the universities of Altdorf and Helmstedt, and his disciple, Christoph Jacob Trew (1695--1769), who became a renowned physician and natural scientist in Nuremberg, communicated in letters to one another over a span of almost forty years. Their correspondence started as a rather asymmetrical dialogue. Over time, however, Heister and Trew came to meet as equals in almost every field of their competence. Their letters reveal a broad spectrum of scientific, organisational, professional, medical and personal issues which formed the basis of a stable and lasting learned correspondence in the age of Enlightenment. PMID- 15889706 TI - [Medical consultation by letter in the 18th century]. AB - Medical consultations by letter are especially abundant in the 18th century; recent research in the history of medicine has focused on this kind of archives, hoping to get a better idea of lay medical culture and medical practice, everyday life of the patient in the early modern period, private experience of suffering, relationships between popular knowledge and medical theories of illness, as well as the major factors of the doctor-patient relationship. However, to interpret them is not an easy nor an univocal task. This article suggests to analyse medical consultations by letter as an elaborate practice, starting from the communicational structure of the material in order to legitimate a two-scale approach, i.e. from the perspective of the healer and the person who is asking for a healing advice. In the first case, we analyse the correspondence of the Bernese physician Albrecht von Haller (1708--1777), and in the second case, the correspondence of the Vaudois physician Samuel-Auguste Tissot (1728--1797), with the aim of developing an approach of systematic comparative research. PMID- 15889707 TI - [The Republic of Letters and French physicians on the eve of the French Revolution: the case of Esprit Calvet]. AB - In the broad Republic of Letters of the second half of the eighteenth century, physicians played an important but singular role. The majority of them were forced to earn their daily bread, so only belonged to the Republic in their leisure hours. Inhabiting a double universe--the everyday world of their profession and the more refined world of their intellectual hobbies--they had to negotiate continually between the two. This liminal position of the citizen physician is recaptured in this article through the example of a physician of the Midi, Esprit Calvet of Avignon (1728--1810). Calvet left a huge correspondence, thanks to which this duality between the practising physician and the citizen of the Republic of Letters can be studied in detail. On the one hand, this is a correspondence between the physician and his patients, augmented by letters between the physician and other physicians on medical topics. On the other hand, it is a correspondence between the physician and other men of science on non medical subjects (archaeology, botany, bibliophily, poetry, etc.). PMID- 15889708 TI - Why, what and how? Editing early modern scientific letters in the 21st century. PMID- 15889709 TI - Transgenic corn for control of the European corn borer and corn rootworms: a survey of Midwestern farmers' practices and perceptions. AB - In 2001, a self-administered questionnaire was sent to 1000 corn, Zea mays L., farmers in each of five states (Illinois, Indiana, Iowa, Minnesota, and Nebraska) to evaluate their perceptions of transgenic corn designed to control the European corn borer, Ostrinia nubilalis (Hubner), and corn rootworms, Diabrotica spp. Respondents returned 1,313 surveys (26.2%). Farmers with small acreages planted a greater portion of their corn (54.5%) with transgenic corn for control of European corn borer than farmers with large farms (39.2%). The majority (75.2%) of farmers use crop rotation to control the corn rootworm. Nine insecticides comprised 92.2% of the commercial soil insecticides used for control of corn rootworm larvae. More than one-third of the farmers in Illinois (33.5%) and Indiana (39.4%) treated first-year corn for corn rootworm, primarily due to western corn rootworm egg laying in soybean, Glycine max (L.). When asked whether they would plant transgenic corn protected against the corn rootworm, 35.0% of farmers responded they would, whereas 40.5% said they were unsure. The two greatest farmer concerns about transgenic corn were the ability to sell harvested grain (59.3%) and additional technology fees (54.8%). Respondents indicated that less farmer exposure to insecticide (69.9%) and less insecticide in the environment (68.5%) were the primary benefits of transgenic corn. Farmers who had no concerns about transgenic corn for rootworm control were more likely to purchase the product (46.8%). The most common refuge-planting options farmers favored were adjacent fields (30.9%) and split fields (29.9%). Farmers (21.1%) observed a yield increase (23.7 bu/ha [9.6 bu/acre]) when using transgenic corn for European corn borer control compared with non-transgenic corn. These data can help in understanding farmers' knowledge and concerns regarding transgenic corn. This information may be of value to guide researchers, extension specialists, and policy makers in designing insect resistance management and integrated pest management programs. PMID- 15889710 TI - Development of genetic sexing strains in Lepidoptera: from traditional to transgenic approaches. AB - The sterile insect technique (SIT) is currently being used for the control of many agricultural pests, including some lepidopteran species. The SIT relies on the rearing and release of large numbers of genetically sterile insects into a wild population. The holokinetic chromosomes of Lepidoptera respond differently to radiation than do species where there is a localized centromere. This difference has enabled a variation of the SIT to be developed for Lepidoptera where a substerilizing dose of radiation is given to the insects before their release with the result that a certain level of sterility is inherited by the F1 offspring. The development of genetic sexing strains for fruit flies, enabling the release of males only, has resulted in enormous economic benefits in the mass rearing and has increased the efficiency of the field operations severalfold. This article outlines Mendelian approaches that are currently available to separate large numbers of males and females efficiently for different lepidopteran species and describes their difficulties and constraints. Successful transgenesis in several lepidopteran species opens up new possibilities to develop genetic sexing strains. The proposal to develop genetic sexing strains described in this article takes advantage of the fact that in Lepidoptera, the female is the heterogametic sex, with most species having aWZ sex chromosome pair, whereas the males are ZZ. This means that if a conditional lethal gene can be inserted into the W chromosome, then all females should die after the application of the restrictive condition. The assumptions made to accommodate this model are discussed, and the advantages to be gained for control programs are elucidated. PMID- 15889711 TI - Evaluation of the neotropical stingless bee Melipona quadrifasciata (Hymenoptera: Apidae) as pollinator of greenhouse tomatoes. AB - The Neotropical stingless bee Melipona quadrifasciata Lepeletier was evaluated for pollinating tomatoes (variety Rodas; long-life hybrid) in greenhouses under plastic and with a hydroponic system and "organic concepts" in Minas Gerais State, Brazil. Flowers not pollinated did not set any fruit. Pollination by bees plus manual pollination did not differ from either bee or manual pollination. Maximum fruit diameter, fruit height, and roundness (quotient between maximum fruit diameter and fruit height) were not significantly different between treatments, but fruit visited by M. quadrifasciata had 10.8% less seeds (dry mass) than manual pollination. This apparently low efficiency of M. quadrifasciata pollination was attributed to the overlap of only 30 min between highest bee foraging activity and highest flower stigma receptivity. Thus, it was concluded that M. quadrifasciata is a feasible pollinator of greenhouse tomatoes because of 1) the observed increase in fruit quality with lower mechanical injury than traditional manual pollination, 2) no significant decrease in fruit size, and 3) high price of such product in the market. Some considerations for sustainable use of M. quadrifasciata as greenhouse pollinator are presented. Although techniques for keeping captive colonies of M. quadrifasciata are currently available, the sole current method for acquiring new colonies is removing them from the forest, and if demand was created for large numbers of colonies for commercial use, techniques for captive rearing must be developed to prevent serious declines in wild populations. PMID- 15889712 TI - Formic acid-based treatments for control of Varroa destructor in a Mediterranean area. AB - Two formic acid autumnal treatments, gel packets (BeeVar formulation) and impregnated paperwick (Liebig-Dispenser), were tested in apiary to evaluate their effectiveness against Varroa destructor Anderson & Trueman and their residues in honey in a Mediterranean region (Sardinia, Italy). Both treatments were efficient in the apiary control of the varroosis, with values of percentage of mite mortality ranging between 93.6 and 100%, without statistical differences between them. The more gradual release of formic acid from the gel application allowed a longer action (2 wk for each treatment) compared with the Liebig-Dispenser (approximately 3d for each treatment). The rate of daily evaporation ranged between approximately 5 and 9 g/d from BeeVar and approximately 26 and 35 g/d from the Liebig-Dispenser, in the first and second treatment, respectively. The total amount of formic acid administered per hive during all the treatment period was approximately 200 g for either treatment. A significantly higher adult bee mortality was recorded in the Liebig-Dispenser-treated hives compared with the BeeVar-treated group. On the contrary, BeeVar treatment produced an interruption of brood reared, whereas the extension of the sealed brood area of the Liebig Dispenser-treated hives was not significantly different from that of the control hives. Neither queen mortality nor robbing activity was observed due to the treatments. Formic acid residues in honey collected in the nest were 3,855 +/- 2,061 and 3,030 +/- 1,624 mg/kg for the BeeVar- and the Liebig-Dispenser-treated hives, respectively. After 21 d from the end of the treatment, the residues fell to 1,261 +/- 1,054 and 794 +/- 518 mg/kg for the honey sampled from the BeeVar and Liebig-Dispenser groups, respectively. PMID- 15889713 TI - Hemolymph juvenile hormone titers in worker honey bees under normal and preswarming conditions. AB - Swarming is an important mechanism by which honey bee, Apis mellifera L., colonies reproduce, yet very little is known about the physiological changes in workers that are preparing to swarm. In this study, we determined the endocrine status of worker honey bees in preswarming colonies and in normal (nonswarming) colonies. Juvenile hormone (JH) titers in worker bees were similar in both groups before queen cells were present, but they became significantly lower in preswarming colonies compared with normal colonies when queen cells occurred in preswarming colonies. The lower JH titers in the preswarming colonies suggest that behavioral development is delayed in these colonies, consistent with previous reports that preswarming colonies have reduced foraging activities. Understanding the endocrine status of bees preparing for swarming will help us to better understand the biology of swarming. PMID- 15889714 TI - Association of beet leafhopper (Hemiptera: Cicadellidae) with a clover proliferation group phytoplasma in Columbia basin of Washington and Oregon. AB - At least 16 taxa of cicadellids and delphacids were tested by polymerase chain reaction (PCR) for the presence of a phytoplasma in the clover proliferation group, designated 16SrVI. Nucleic acid extracts from individual insects or groups of 5-10 were tested using PCR primers designed from the DNA sequence of 16S-23S rRNA or ribosomal protein genes of the pathogen. The beet leafhopper, Circulifer tenellus (Baker), was most often associated with the phytoplasma, with approximately 16% of the insects testing positive. The phytoplasma was occasionally found associated with Ceratagallia spp. Leafhopper species that were not associated with the phytoplasma included Macrosteles spp., Dikraneura spp., Colladonus montanus (Van Duzee), Circulifer geminatus (Van Duzee), Ballana spp., Amplysellus spp., Paraphlepsius spp., Texananus spp., Balclutha spp., Latalus spp., Erythroneura spp., Exitianus exitiosus (Uhler), and unidentified delphacids. The detected phytoplasma was similar to, or synonymous with, the beet leafhopper-transmitted virescence agent that is associated with the potato purple top disease in the Columbia Basin region of Washington and Oregon. This is in contrast to the phytoplasma associated with potato purple top disease in Mexico that is related to aster yellows (group 16SrI). The association of the group 16SrVI phytoplasma almost exclusively with the beet leafhopper suggests that this insect is the major vector of the phytoplasma in this region. PMID- 15889715 TI - Hot water immersion for surface disinfestation of Maconellicoccus hirsutus (Homoptera: Pseudococcidae). AB - Mealybug Maconellicoccus hirsutus (Green) adults, nymphs, crawlers, and eggs were tested for their susceptibility to hot water immersion at 47, 48, and 49 degrees C. Eggs inside ovisacs were found most tolerant with prolonged survival compared with other stages at all temperatures. Ovisacs required an average of 1.38, 1.46, and 1.62 times longer treatment duration than adults, nymphs, and crawlers, respectively, for 99.9% predicted mortality at 47, 48, and 49 degrees C. Lethal time estimations were calculated from inverse predictions of regressions derived from logit-transformed data as well as those created using a kinetic model. LT 99.9 estimations were 47.0, 21.2, and 11.9 min at 47, 48, and 49 degrees C, respectively, by using regressions with logit transformations. The kinetic model predictions were 43.9, 19.6, and 11.1 min at 47, 48, and 49 degrees C, respectively. During the study no emergence from eggs inside ovisacs was found after treatments of 52, 24, and 14 min at 47, 48, and 49 degrees C, respectively. Results from this study provide efficacious temperature-time treatments. PMID- 15889716 TI - Potential of temperature, controlled atmospheres, and ozone fumigation to control thrips and mealybugs on ornamental plants for export. AB - Ozone (O3) fumigation is a potential quarantine treatment alternative for controlling stored-product pests and surface insect pests on fresh agricultural commodities. We explored the effects of temperature, treatment time, controlled atmospheres, and vacuum in combination with O3 to control two important pests of ornamental crops: western flower thrips, Frankliniella occidentalis (Pergande), and longtailed mealybug, Pseudococcus longispinus Targioni Tozzetti. Treatment parameters tested were O3 concentrations from 0 to 3,800 ppm, treatment durations were from 30 to 120 min, vacuums were from 0 to 0.41 bar below ambient, temperatures were from 32.2 to 40.6 degrees C, and controlled atmospheres were composed primarily of nitrogen, carbon dioxide, or breathing air [BA]. Treatment efficacy was enhanced by higher O3 concentration and temperature, lower oxygen, and longer treatment times. Reduced pressure was not an important factor. Mealybugs were more difficult to kill than thrips. A 30-min treatment of O3 at approximately 200 ppm in 100% CO2 at 37.8 degrees C killed 47.9 and 98.0% of mealybugs and adult female thrips, respectively. All of the ornamentals tested were damaged to some degree by O3 treatments. However, crops with thick leaves such as orchids exhibited little damage, and the waxy portions of certain flowers were not damaged. The results suggest that O3 has potential as a quarantine treatment to control thrips and mealybugs on selected commodities. PMID- 15889717 TI - DNA diagnostics to identify internal feeders (Lepidoptera: Tortricidae) of pome fruits of quarantine importance. AB - A diagnostic polymerase chain reaction (PCR) method is presented for differentiating among the North American internal apple-feeding pests codling moth, Cydia pomonella (L.); oriental fruit moth, Grapholita molesta (Busck); lesser appleworm, Grapholita prunivora (Walsh); and cherry fruitworm, Grapholita packardi Zeller. An approximately 470-bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in three to six specimens of each species. Consistent and diagnostic differences were observed among the species in two regions of COI from which forward and reverse primers were designed to amplify a 112-116-bp segment of the gene. The primer sets were used to selectively amplify DNA from specimens of diverse geographic origin for each corresponding target species. Protocols were adapted for conventional and quantitative PCR, the latter being substantially faster. The method was validated as a decision-making tool for quarantine identifications for Mexico by representatives of their phytosanitary agency (Sanidad Vegetal). The method can facilitate identification of intercepted internal feeding Lepidoptera in apple and pear for many other importing nations. PMID- 15889718 TI - Competitive interactions among four pest species of earth mites (Acari: Penthaleidae). AB - Earth mites are major winter pests of a variety of crops and pastures in southern Australia. Competition between four earth mite species was investigated using field and shadehouse experiments. The influence of different plant hosts on the frequency and intensity of competitive interactions also were examined. This information is important, because control attempts that eradicate one species of mite could be directly followed by an increase in abundance of another earth mite species. There were strong effects of intraspecific competition on the reproductive rate of species, while interspecific interactions between Halotydeus destructor (Tucker) and Penthaleus species and between the three Penthaleus species also were detected. Competitive abilities were altered on the different plant types. On pasture, the competitive advantage swayed between Penthaleus major (Duges), H. destructor, and Penthaleus falcatus (Qin & Halliday). Penthaleus sp. x was the strongest competitor in a mixture of wheat, Triticum aestivum (L.), and oats, Avena sativa (L.), whereas on canola, Brassica napus (L.), and bristly ox-tongue, Picris echioides (L.), P. falcatus, and H. destructor were superior competitors. These results suggest that competition is a strong force influencing the abundance of earth mites in the field and that host plant factors are important in shaping the type of interactions. This highlights the importance of identifying mite species when considering control options and suggests that effective control recommendations need to be developed for each individual species. PMID- 15889719 TI - Male oriental fruit moth response to a combined pheromone-based attracticide formulation targeting both oriental fruit moth and codling moth (Lepidoptera: Tortricidae). AB - Combined attracticide formulations targeting Oriental fruit moth, Grapholita molesta (Busck), and codling moth, Cydia pomonella (L.), were tested in a field trapping experiment. Capture of male codling moths in traps baited with the combined formulation was reduced compared with traps baited with the codling moth formulation alone, whereas capture of male Oriental fruit moth was increased compared with traps baited with the Oriental fruit moth formulation alone. Subsequent wind tunnel experiments showed that a single locus of the mixed attracticide formulation or close parallel presentation of the two formulations enhanced source contact by male Oriental fruit moths but did not influence earlier behaviors. However, the two formulations presented in a serial arrangement to Oriental fruit moth males in the wind tunnel resulted in enhanced lock-on, upwind flight, and source contact behaviors. In addition, male Oriental fruit moths remained on mixed pheromone droplets of the paste matrix longer than on droplets of the Oriental fruit moth formulation alone. The increased time spent on the mixed droplet was correlated with a more rapid poisoning and a greater proportion of poisoned males compared with males exposed to the Oriental fruit moth attracticide alone. These results demonstrate that a combined attracticide formulation will have different effects on each of the targeted species. It is anticipated that, due to decreased attractiveness, a combined formulation would be less effective against the codling moth. However, a mixed formulation, due to increased attractiveness and toxicity, could be more effective against the Oriental fruit moth under field conditions. PMID- 15889720 TI - Plodia interpunctella (Lepidoptera: Pyralidae): spatial relationship between trap catch and distance from a source of emerging adults. AB - The Indianmeal moth, Plodia interpunctella (Hubner), was selected as a representative stored-product moth to test the validity of contour mapping of trap catch for pest monitoring in warehouses and retail stores. Three experiments, each replicated five times, were conducted in a 3.2 by 9.0-m aluminum shed. Each experiment involved placing pupae at a single release point (source) and recording the numbers of emerging adult males captured after 24, 48, and 72 h in each of four pheromone-baited sticky traps. The experiments differed only with respect to the point of release. Distribution of trap catch reflected the general distribution of moths in the shed; and consecutive contour maps tracked their dispersal from the source. As emergence and dispersal progressed, cumulative trap catch increased throughout the shed, but it remained highest near the source. The observed spatial patterns of trap catch relative to sources of infestation and the inverse relationship of trap catch to distance from a source support the validity of contour mapping as a means of monitoring stored-product moths and locating foci of infestation. The relationship between trap catch and distance from a source of infestation was well described by two-parameter exponential decay, both in P. interpunctella and in the previously studied Lasioderma serricorne (F.). Analysis of data from retail pet stores also showed exponential decline in trap catch of P. interpunctella with distance from centers of infestation. PMID- 15889721 TI - Effects of exposure to pheromone and insecticide constituents of an attracticide formulation on reproductive behavior of oriental fruit moth (Lepidoptera: Tortricidae). AB - The effect of exposure to both the pheromone and insecticide constituents of an attracticide formulation on subsequent pheromonal response of male oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), was tested in several wind tunnel bioassays. Male response to the attracticide formulation was significantly reduced in all behavioral categories, including source contact 1 h after sublethal exposure (both by voluntary contact in the wind tunnel and forced application in the laboratory) to the attracticide formulation containing inert ingredients, pheromone, and insecticide. Sublethal exposure to the attracticide formulation in the laboratory (forced application) 24 h before the bioassay resulted in a significantly lower proportion of males subsequently responding to attracticide droplets in the wind tunnel. However, voluntary contact of male moths with the toxic formulation in the wind tunnel had no effect on subsequent response 24 h later. Exposure of males to different constituents of the attracticide formulation demonstrated that both pheromone and insecticide exerted effects on subsequent male pheromonal response. Exposure to the formulation containing the inert ingredients plus the pheromone (no insecticide) significantly reduced male behavioral responses to an attracticide droplet in the wind tunnel 1 h but not 24 h after exposure, compared with males treated with inert ingredients alone. Response to attracticide droplets was further reduced by exposure to the entire attracticide formulation containing inert ingredients, pheromone and insecticide at both 1 and 24 h postexposure. Similarly, males exposed to inert ingredients plus pheromone were less likely to orient to female produced plumes 1 h but not 24 h after exposure than males treated with inert ingredients alone. Response to female-produced plumes was further reduced at 1 h but not at 24 h after exposure to the entire attracticide formulation. Mating success of males was significantly reduced by exposure to the entire attracticide formulation but not to the formulation without insecticide when placed with females 1 and 24 h postexposure. These findings suggest that sublethal poisoning of males exposed to the attracticide formulation will enhance the effectiveness of this formulation under field conditions. PMID- 15889722 TI - Mushroom host influence on Lycoriella mali (Diptera: Sciaridae) life cycle. AB - Lycoriella mali Fitch (Diptera: Sciaridae) infests mushroom crops early in the crop cycle. Recent observations in mushroom houses indicated a difference in emergence time and size of adult L. mali developing on various strains of commercial mushrooms. Samples of adult flies from isolated mushroom houses growing Portabella mushrooms were significantly heavier then those from oyster mushroom houses, whereas flies from shiitake mushroom houses were lightest in weight. Flies collected from isolated Portabella mushroom houses were reared on four strains and species of Agaricus and Pleurotus mushrooms. After the adults emerged, females were weighed, mated, and allowed to oviposit. The number of eggs laid increased as the weight of the female increased. Flies collected from isolated Portabella mushroom houses were reared on eight strains and species of mushrooms. Flies were reared for four generations on each host mushroom mycelium then switched to different host mushrooms. Overall, the hybrid strain of Agaricus bisporus (Lange) Imbach (Agaricales: Agaricomycetideae) was the most favorable host for L. mali, whereas the wild strain of A. bisporus was the least favorable host. Mushroom hosts influence developmental time, survivorship, weight, and reproduction of L. mali. PMID- 15889723 TI - Quantifying pepper maggot (Diptera: Tephritidae) oviposition and infestation over time. AB - In 1998 and 1999, pepper maggot flies, Zonosemata electa (Say), were excluded from pepper plants with row covers for 0-6 wk after initial oviposition was detected. The proportion of oviposition attempts (scars) and larval infestation that occurred each week and the cumulative oviposition and infestation over time were determined. Due to environmental factors, initial oviposition was delayed, and the numbers of oviposition scars on fruit and flies caught on traps were lower in 1998 than in 1999. The proportions of scarred or infested fruit were low (< 8% of total) during the first 7 d after the pest was detected each year. Oviposition peaked 8-14 d after beginning in 1998, but not until 21-28 d after the initial scar was detected in 1999. Because systemic insecticides tend to provide control of adults and most hatching larvae, it may be possible to delay the initial insecticide application for up to a week after oviposition commences and still minimize fruit damage. There was no association with either year between the proportion of flies captured on traps and the proportion of oviposition scars on fruit the same week or 1 and 2 wk later. PMID- 15889724 TI - Biochemical stress indicators of greater wax moth exposure to organophosphorus insecticides. AB - Although acetylcholinesterase (AChE) is the primary target of organophosphorus insecticides (OPs), increasing evidence regarding their secondary effects suggests that OPs disturb homeostasis of insects by generating free radical intermediates that trigger lipid peroxidation. We therefore investigated alterations in lipid peroxidation product, malondialdehyde (MDA) content, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, in conjunction with AChE activity as biochemical stress indicators in greater wax moth, Galleria mellonella (L.) larvae for OPs methyl parathion (MP) and ethyl parathion (EP). The effects of MP and EP were first investigated by rearing the young larvae on an artificial diet containing 0.01, 0.1, 1, 10, and 100 ppm of each insecticide. Second, the mature larvae were injected with 0.05, 0.5, 5, 50, and 500 ng of insecticides for determining the changes in biochemical stress responses. The diet with lowest level of MP significantly decreased the activities of all measured enzymes, whereas it increased MDA content. However ALT and AST were significantly higher in the larvae reared with the diet with high levels of MP than in control larvae. All tested levels of MP resulted in a decrease in AChE activity. The lowest level of EP in diet (0.01 ppm) significantly increased ALT activity, whereas it reduced that of AChE. This insecticide at 0.1 ppm resulted in reduced AST activity, but 1 ppm in diet elevated AST activity and MDA content. EP at 0.1 ppm and higher levels in the diet reduced ALT activity. All dietary EP levels significantly decreased AChE activity. ALT, AST, and AChE were lower in larvae fed with the diet containing 100 ppm ethyl parathion compared with larvae on control diet. MP at 50 ng per larva increased ALT and AST activities from 35.42 +/- 0.74 and 26.34 +/- 0.83 to 203.57 +/- 1.09, and 122.90 +/- 1.21 U/g, respectively, when the mature larvae were injected. All injected doses of EP dramatically reduced both ALT and AST activities, but only the lowest and highest levels of this insecticide decreased AChE activity. The lowest level of this insecticide also significantly increased MDA content in larvae. High levels of both insecticides increased MDA content. We observed a significant higher increase in MDA content in the larvae reared with 10 ppm EP (102.16 +/- 1.57 nmol/g protein) than the control group (30.28 +/- 1.42 nmol/g protein). These results suggest that OPs caused the metabolic and synaptic dysfunctions in greater wax moth and alter its biochemical physiology in response to oxidative stress. PMID- 15889725 TI - Influence of honey bee (Hymenoptera: Apidae) density on the production of canola (Crucifera: Brassicacae). AB - Pollination is an essential step in the seed production of canola, Brassica napus L. It is achieved with the assistance of various pollen vectors, but particularly by the honey bee, Apis mellifera L. Although the importance of pollination has been shown for the production of seed crops, the need to introduce bee hives in canola fields during flowering to increase oil seed yield has not yet been proven. With the purpose of showing this, hives of A. mellifera were grouped and placed in various canola fields in the Chaudiere-Appalaches and Capitale Nationale regions (nine fields; three blocks with three treatments; 0, 1.5, and 3 hives per hectare). A cage was used to exclude pollinators and bee visitations were observed in each field. After the harvest, yield analyses were done in relation to the bee density gradient created, by using pod set, number of seeds per plant, and weight of 1000 seeds. Results showed an improvement in seed yield of 46% in the presence of three honey bee hives per hectare, compared with the absence of hives. The introduction of honey bees contributed to production and consequently, these pollinators represented a beneficial and important pollen vector for the optimal yield of canola. PMID- 15889726 TI - Effect of aging on pheromone emission from a commercial beet armyworm (Lepidoptera: Noctuidae) lure and trap efficiency. AB - The effect of aging on a commercial pheromone-based lure for attractiveness to male adult beet armyworms, Spodoptera exigua (Hubner), was assessed in terms of trapping efficiency and volatile emissions of two key components: (Z,E)-9,12 tetradecadienyl acetate and (Z)-9-tetradecen-1-ol. In field assays conducted in the Lower Rio Grande Valley, Texas, for 9 wk (June-August 2000) and for 8 wk (March-May) in Tamaulipas, Mexico, 0-1-wk-old lures collected greater than four fold more male beet armyworm adults than lures that had been aged for 3-4 wk. Using solid phase microextraction and gas chromatographic analysis of the volatiles, mean (Z,E)-9,12-tetradecadienyl acetate emission declined by 32% after 5 wk of aging in a ventilated environmental chamber at 29.4 degrees C, 3.5% RH, to simulate subtropical summers. Mean (Z)-9-tetradecen-1-ol emission was reduced by 62% after 4 wk of aging. Under the tropical and subtropical conditions of this study, the capacity of the lure to attract moths into the trap declined after aging for 3 wk. The blend ratio (Z,E) -9,12-tetradecadienyl acetate: (Z)-9 tetradecen-1-ol declined 29% when the lure was aged for 5 wk. This study will assist in the development of a standard trapping system for assessing adult beet armyworm populations in the tropical and subtropical conditions to which the species is endemic. PMID- 15889727 TI - Characterization of the spatial dispersion of corn root injury by corn rootworms (Coleoptera: Chrysomelidae). AB - The dispersion of root injury to Zea mays L. by corn rootworm, Diabrotica spp., larvae was characterized using geostatistics to determine the appropriate sampling distance to obtain independent estimates of root injury. We also investigated the effect of the root injury levels on the spatial dependence of root injury at three different spatial scales: large-scale study with 25-m sampling distance in three, 8-ha fields; moderate-scale study with 0.2-m sampling distance in six, 2-ha fields; and small-scale study with exhaustive sampling in eight, 12-m(2) plots within cornfields. In the large-scale study, a nugget model best fit the semivariograms, indicating the dispersion of root injury exhibited a random pattern. In the moderate-scale study, all the semivariograms were best fit with an exponential model, and 68 to 96% of the variability was explained by spatial dependence, suggesting a high degree of spatial aggregation of corn root larval injury. The ranges of semivariograms fell within < 4 m, except one field that had the range of 14.7 m. In the small-scale study, the Gaussian, exponential, or spherical models fit the semivariograms best and 50 to 89% of the variability was explained by spatial dependence, suggesting spatial aggregation of root injury. The ranges of semivariograms were from 0.35 to 1.04 m. In the small-scale study, the degree of spatial dependence increased significantly (P < 0.05) as the average root injury increased within the range of root injury observed in the study. These results suggest that the minimum distance between root samples in small-plot insecticide trials should be > 1 m within a row to obtain unbiased estimates of root injury. PMID- 15889728 TI - Influence of maize/lablab intercropping on lepidopterous stem borer infestation in maize. AB - Lepidopterous stem borers seriously affect production of maize, Zea mays L., in sub-Saharan Africa. Intercropping maize with legumes such as lablab, Lablab purpurens (L.), is one of the effective systems to control stem borers. Sole culture maize and maize/lablab intercrop system of different lablab densities were planted at two locations to investigate the effects of intercrop system on incidence and severity of stem borers with particular reference to Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae). Stem borer infestation was found to be more severe in sole culture maize than maize in maize/lablab intercrop. There was a significantly negative relationship between lablab densities and maize grain yields, suggesting a possible competition for resources between the two crops. It was concluded that density of lablab and date of planting of lablab in maize/lablab intercropping have significant affects on stem borer populations and maize grain yields. PMID- 15889729 TI - Influence of three resistance sources in winter wheat derived from TAM 107 on yield response to Russian wheat aphid. AB - A study to determine yield response to the Russian wheat aphid, Diuraphis noxia (Mordvilko), was conducted during the 1997-1998 and 1998-1999 growing seasons at three eastern Colorado locations, Akron, Fort Collins, and Lamar, with three wheat lines containing either Russian wheat aphid-resistant Dn4 gene, Dn6 gene, or resistance derived from PI 222668, and TAM 107 as the susceptible control. Russian wheat aphids per tiller were greater on TAM 107 than the resistant wheat lines at the 10x infestation level at Fort Collins and Akron in 1999. Yield, seed weight, and number of seeds per spike for each wheat line were somewhat affected by Russian wheat aphid per tiller mainly at Fort Collins. The infested resistant wheat lines harbored fewer Russian wheat aphids and yielded more than the infested susceptible wheat lines. Wheat lines containing the Dn4, Dn6, and PI 222668 genes contain different levels of antibiosis or antixenosis and tolerance. Although differences existed among sites and resistance, there is a benefit to planting resistant wheat when there is a potential for Russian wheat aphid infestations. PMID- 15889730 TI - European corn borer (Lepidoptera: Crambidae) effects on qualitative traits of high oil content corn grown in Kentucky. AB - The European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae), is one of the most important pests of corn, Zea mays L., because it consistently causes high loss of yield. A study was conducted in 2000-2002 at field sites in central and western Kentucky to investigate whether infestation by O. nubilalis differentially affects the production of high-oil corn compared with traditional field corn. Statistical differences in grain weight and percentage of oil content between the five infestation levels were significant at both locations and for all years. Average grain yield was reduced by 0.40% and average oil concentration by 0.011% for each 1% of damaged plants, and there was a strong correlation (0.76) between leaf damage ratings (i.e., Guthrie scale) and yield reduction. In general, corn planted at the early planting date tended to have a higher yield (grain weight) and oil content. PMID- 15889731 TI - Assessment of cone-damaging insects in a Swedish spruce seed orchard and the efficacy of large-scale application of Bacillus thuringiensis variety aizawai x kurstaki against lepidopterans. AB - In a 2-yr study, we investigated the efficacy of large-scale application of the biological insecticide Bacillus thuringiensis variety aizawai x kurstaki (Btk) in a Swedish spruce, Picea abies L. (Karst.), seed orchard for controlling damage caused by four lepidopteran species: Dioryctria abietella Den et. Schiff. (Pyralidae), Eupithecia abietaria Gotze, E. analoga Djakonov (Geometridae), and Cydia strobilella (L.) (Tortricidae). The frequencies of these species, and Strobilomyia anthracina Czerny (Diptera: Anthomyiidae), were regularly monitored throughout the vegetative growth season to map their temporal distribution patterns and to quantify occurrences of species that may have been present in the cones at some stage during the season but migrated before the final sampling. This investigation revealed that E. abietaria occurred in similar numbers to D. abietella and has probably been overlooked as a potentially serious pest in spruce seed orchards in Sweden. To determine the number, timing, and rate of Btk required to control the lepidopterans, spraying was conducted at different phases of flowering and cone development, and three rates of Btk were applied. The Btk treatment reduced cone damage caused by D. abietella and Eupithecia spp. by one half in 2002, a year with an intermediate number of cones, but the effect was weaker in 2003, when the cone crop was low. Damage caused by C. strobilella was not affected by the treatment. The different rates of Btk application did not affect the results, and repeated spraying seemed to be effective during 2002 but not in 2003. PMID- 15889732 TI - Evaluation of biological and biorational control tactics for suppression of Nantucket pine tip moth damage in Virginia pine Christmas trees. AB - A study was performed to evaluate the potential of the egg parasitoid Trichogramma exiguum Pinto and Platner (Hymenoptera: Trichogrammatidae), the newly registered insect growth regulator (IGR) tebufenozide (Confirm), and a modified spray technique (top whorl only pesticide application) for suppression of Nantucket pine tip moth, Rhyacionia frustrana (Comstock) (Lepidoptera: Tortricidae), damage in Virginia pine, Pinus virginiana Mill., Christmas trees. Augmentative releases of T. exiguum failed to increase parasitism levels in release plots compared with controls, and significant reduction in tip moth damage did not occur. High predation levels on released T. exiguum may have contributed to the failure of parasitoid augmentations. Whole-tree and top whorl tebufenozide treatments provided significantly greater damage control than corresponding applications of acephate (Orthene), a commonly used pesticide in Christmas trees. Damage to trees receiving whole-tree chemical applications did not differ significantly from trees receiving top whorl treatments for most measurements of damage. Top whorl chemical treatments resulted in a 67% reduction in time required for application and a 70% reduction in pesticide used. PMID- 15889733 TI - Stage-specific control of grape berry moth, Endopiza viteana (Clemens) (Lepidoptera: Tortricidae), by selective and broad-spectrum insecticides. AB - The insect growth regulators (IGRs) tebufenozide and methoxyfenozide and the broad-spectrum insecticides azinphosmethyl, carbaryl, and fenpropathrin were compared for their activity against adult, egg, and larval stages of the grape berry moth, Endopiza viteana (Clemens) (Lepidoptera: Tortricidae), under laboratory and vineyard conditions. Adult mortality was not affected by exposure to field-equivalent rates of tebufenozide or methoxyfenozide on grape clusters, whereas all the broad-spectrum compounds significantly reduced adult survival, compared with the untreated controls. Surviving adult moths laid significantly more eggs on berries treated with the IGRs than on berries treated with any of the broad-spectrum insecticides. Survival of these eggs through to late larval and pupal stages was significantly lower on methoxyfenozide-treated grapes than on untreated grapes, and no pupae were found when grapes were treated with azinphosmethyl or fenpropathrin. Neither of the growth regulator insecticides limited egg eclosion or larval development by E. viteana when insecticides were applied before egg laying, whereas broad-spectrum insecticides were effective against both eggs and neonates at this timing. When applied after egg eclosion, all insecticide treatments significantly reduced survival of grape berry moth larvae. Under vineyard conditions, berries with 1-d-old residues of tebufenozide or methoxyfenozide received more E. viteana eggs than berries treated with broad spectrum compounds. After aging for 7 or 14 d, no significant effects on E. viteana survival were detected among treatments. Whereas broad-spectrum insecticides provide control of multiple life stages of E. viteana, integration of tebufenozide or methoxyfenozide into vineyard management programs for control of this pest will be most successful if applications are timed for egg hatch. PMID- 15889734 TI - Effect of a plant growth regulator prohexadione-calcium on insect pests of apple and pear. AB - The effect of prohexadione-calcium, a plant growth regulator that inhibits gibberellin metabolism, on Cacopsylla pyricoloa (Foerster) in pear trees, and Choristoneura rosaceana (Harris) and Aphis spireacola Patch, in apple trees was studied. C. pyricoloa and A. spireacola populations were significantly reduced in prohexadione-calcium-treated pear and apple, respectively. Insecticide control of both pests with imidacloprid was synergized in treatments with prohexadione calcium. In apples treated with prohexadione-calcium, there was a significant reduction in the number of C. rosaceana shelters per tree and amount of fruit injury at harvest attributable to the C. rosaceana. There was an additive effect when tebufenozide was used to control C. rosaceana in trees treated with prohexadione-calcium. Prohexadione-calcium significantly reduced vegetative growth in both pears and apples. Synergistic and additive treatment effects of prohexadione-calcium and pesticides used in this study may be due to better penetration and coverage of pesticides due to reduced foliar growth or to changes in the nutritional quality of the host plants. PMID- 15889735 TI - Protection of fruit against infestation by apple maggot and blueberry maggot (Diptera: Tephritidae) using compounds containing spinosad. AB - Two insecticide formulations containing the naturalyte insecticide spinosad, GF 120 Fruit Fly Bait and SpinTor 2 SC, were compared for control of apple maggot, Rhagoletis pomonella (Walsh), and blueberry maggot, Rhagoletis mendax Curran. In 2002 and 2003, larval infestation in blueberries and apples was significantly lower in plots treated with GF-120 (spinosad bait) or SpinTor than in untreated control plots. Fruit fly infestation in apples was reduced by 67% in 2002 after weekly application of GF-120 for 6 wk. Six weeks of GF-120 treatment reduced infestation in blueberries by 85% in 2002 and 98% in 2003. Plots treated weekly with the bait component of GF-120 for 6 wk had significantly higher infestation of blueberry maggot larvae compared with untreated plots in 2002. Observations of wild R. mendax flies revealed that similar numbers of flies landed on blueberry foliage treated with spinosad bait, the bait component alone, or water droplets. However, flies on spinosad bait and bait treated plants spent significantly more time within 5 cm of the treatment droplets compared with control (water) droplets. Overall, the results demonstrate a high degree of efficacy of baited spinosad formulations against these key pests of temperate fruit and suggest that GF-120 is an arrestant for foraging flies. PMID- 15889736 TI - Effects of neem and spinosad on Ceratothripoides claratris (Thysanoptera: Thripidae), an important vegetable pest in Thailand, under laboratory and greenhouse conditions. AB - Toxicity of three biopesticides, i.e., two neem products and spinosad, was determined on foliage-dwelling life stages of Ceratothripoides claratris (Shumsher) (Thysanoptera: Thripidae), a major thrips pest on tomatoes, Lycopersicon spp., in central Thailand. Direct and residual contact toxicities of NeemAzal-TS (1% azadirachtin) and systemic activity of NeemAzal-MD 5 (5% azadirachtin) affected the survival of first larval stage (L1) in a concentration dependent manner. However, neither second larval stage (L2) nor adult survival was influenced by both neem products. On the contrary, spinosad caused 100% mortality in both larval stages and adults of C. claratris regardless of the concentrations tested. No strong ovicidal effects were detected in three different age groups of eggs (i.e., 1, 2, and 3 d old) topically treated with both NeemAzal-TS and spinosad. Residual toxicity was highest with fresh residues of NeemAzal-TS compared with 1-, 3-, 5-, and 7-d-old residues and in general was higher under laboratory than greenhouse conditions. Irrespective of the age of the spray residues, spinosad always caused 100% mortality in larvae and adults. Strongest systemic effects were observed in L1 larvae 1 d after soil drenching with NeemAzal-MD 5 at the highest concentration tested. Foliar and soil applications of NeemAzal-TS and NeemAzal-MD 5, respectively, did not cause any oviposition deterrent effects. PMID- 15889737 TI - Hymenopteran parasitoids of diamondback moth (Lepidoptera: Ypeunomutidae) in northern Thailand. AB - Larvae of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Ypeunomutidae), cause severe economic damage to cabbage, Brassica oleracea L. variety capitata (Brassicaceae) and related vegetables in Thailand. Overuse of broad-spectrum insecticides for diamondback moth control is a serious problem and has obscured the contributions of indigenous parasitoids. Our objectives were to identify indigenous diamondback moth parasitoids in northern Thailand and to assess their potential for natural control. Six parasitoid species were reared from diamondback moth larvae and pupae collected in 1990 and in 2003-2004. These included the larval parasitoid Cotesia plutellae Kurdjumov (Braconidae), a larval pupal parasitoid Macromalon orientale Kerrich (Ichneumonidae), and pupal parasitoids Diadromus collaris Gravenhorst (Ichneumonidae) and Brachymeria excarinata Gahan (Chalcididae). Single specimens of Isotima sp. Forster (Ichneumonidae) and Brachymeria lasus Walker (Chalcididae) also were reared from diamondback moth hosts. C. plutellae was the dominant larval parasitoid and was often reared from host larvae collected from fields sprayed regularly with insecticides; parasitism ranged from 14 to 78%. Average parasitism by M. orientale was only 0.5-6%. Parasitism of host pupae by D. collaris ranged from 9 to 31%, whereas B. excarinata pupal parasitism ranged from 9 to 25%. An integrated pest management (IPM) protocol using simple presence-absence sampling for lepidopterous larvae and the exclusive use of Bacillus thuringiensis (Bt) or neem resulted in the highest yields of undamaged cabbage compared with a control or weekly sprays of cypermethrin (local farmer practice). IPM programs focused on conservation of local diamondback moth parasitoids and on greater implementation of biological control will help alleviate growing public concerns regarding the effects of pesticides on vegetable growers and consumers. PMID- 15889738 TI - Impact of twospotted spider mite (Acari: Tetranychidae) on growth and productivity of glasshouse cucumbers. AB - Experiments were conducted to assess the damage of the glasshouse cucumber by twospotted spider mite, Tetranychus urticae Koch, and to investigate when the economic yield begins to decrease after T. urticae infestations. To assess the damage, dry matter partitioning in the cucumber plant was quantified and plant growth analyses were conducted at four different T. urticae infestation levels. T. urticae infestations decreased leaf productivity by reducing the total number of leaves per plant. Approximately 14% reductions of total leaf areas could result in significant yield loss. The decreased leaf productivity by T. urticae feeding caused biomass reductions and altered the pattern of dry matter partitioning in the plant; damaged plants accumulated more dry matter in the leaf, and partitioning of dry matter to fruits was hindered. The economic yield of cucumber began to significantly decrease as early as 4 wk after heavy mite infestations. This study also showed the seasonal differences in T. urticae cucumber damage interactions among mite infestation levels. PMID- 15889739 TI - Effects of sublethal concentrations of Bacillus thuringiensis on larval development of Sesamia nonagrioides. AB - In the Mediterranean Basin, developed diapausing larvae of the borer Sesamia nonagrioides Lefebvre are found consistently during autumn in transgenic corn, Zea mays L., expressing Bacillus thuringiensis Berliner (Bt) toxins derived from event 176. These larvae can feed on sublethal concentrations of Bt toxins because the expression of this corn toxin declines after flowering. To evaluate the possible consequences of this phenomenon, the effect of sublethal concentrations of the natural toxin producer, B. thuringiensis variety kurstaki (Dipel DF) on the performance of S. nonagrioides in the laboratory was examined. Larvae treated with Dipel DF showed higher mortality, longer developmental time, extra molts, and higher sensitivity to critical daylength for diapause induction than the untreated larvae. Accordingly, diapausing larvae collected in October and February in a Bt cornfield exhibited a higher number of supernumerary molts before pupating and longer diapause development than larvae collected in a non-Bt field at the same time. These results demonstrate that deployment of Bt corn in the Mediterranean may affect both the borer performance and the development of Bt resistance in addition to direct mortality caused by the Bt toxin. PMID- 15889740 TI - Directional change in tunneling of subterranean termites (Isoptera: Rhinotermitidae) in response to decayed wood attractants. AB - Wood discs decayed with brown rot fungi and polymer discs impregnated with acetone extract of decayed wood were embedded in sand of a two-dimensional foraging arena to evaluate their attractant potential in directing termite tunnels toward them. Termites were released near one arena corner and were guided to follow the physical guideline of the arena edges. In the absence of the attractants, termites generally formed a relatively linear tunnel along the edges. When decayed wood discs or treated polymer discs were placed in wet sand near one side of the arena, termite tunnels departed from the arena edges and were oriented toward them. The attraction distance ranged from 12 to 18 cm. The attractant properties were most likely water soluble and permeated through wet sand to cause termites to change their orientation. The results demonstrated that when such attractants are placed near a bait station in the field, they may be used to direct termite foraging toward the station. PMID- 15889741 TI - Attraction of subterranean termites (Isoptera) to carbon dioxide. AB - Subterranean termites, Reticulitermes spp., were attracted to carbon dioxide (CO2) in laboratory and field tests. In behavioral bioassays, Reticulitermes flavipes (Kollar), Reticulitermes tibialis Banks, and Reticulitermes virginicus Banks were attracted to CO2 concentrations between 5 and 50 mmol/mol. In further bioassays, R. tibialis and R. virginicus were attracted to the headspace from polyisocyanurate construction foam that contained 10-12 mmol/mol CO2. In soil bioassays in the laboratory, more termites foraged in chambers containing CO2 generating formulations than in unbaited control chambers. In field tests, stations containing CO2-generating baits attracted R. tibialis away from wooden fence posts at rangeland sites in Colorado. For all of the CO2 formulations tested, termites foraged in significantly more bait stations at treatment fenceposts than in bait stations at the control fenceposts. By the end of the 8 wk study, the number of bait stations located by termites at treatment fenceposts ranged from 40 to 90%. At control fenceposts, termites foraged in only a single station and the one positive station was not located by termites until week 5 of the study. At treatment fenceposts, termites foraged equally in active stations (containing a CO2-generating bait) and passive stations (with no CO2-generating bait), indicating that bait stations may benefit passively from a proximal CO2 source in the soil. CO2 used as an attractant in current baiting systems could improve their effectiveness by allowing earlier exposure of termites to an insecticide. PMID- 15889742 TI - Efficacy of simulated barrier treatments against laboratory colonies of Pharaoh ant. AB - Five selected insecticides were applied to four substrates and evaluated in laboratory studies for repellency and toxicity against the Pharaoh ant, Monomorium pharaonis (L.). We tested both repellent and nonrepellent formulations on outdoor (concrete and mulch) and indoor (ceramic and vinyl) substrates. Repellency was evaluated using a behavioral bioassay in which colonies were given a choice to leave the treated zone and move into empty nests provided in the untreated zone. We used a novel experimental design whereby ants walked on a Slinky coil suspended from a metal support frame, thus permitting a long foraging distance with a minimum use of space and resources. Cypermethrin, a repellent pyrethroid insecticide, resulted in colony budding, although the response was delayed. Toxicity of insecticides was evaluated as worker, queen, and brood mortality. The most effective treatment was fipronil, which provided 100% reduction in pretreatment activity by 2 d posttreatment on both concrete and mulch. Chlorfenapyr was highly effective on both outdoor and indoor substrates. Significant substrate effects were observed with insecticides applied to nonabsorbent substrates (ceramic tile), which performed better than insecticides applied to absorbent substrates (vinyl tile). Other highly absorbent materials (mulch and concrete), however, did not reduce insecticide efficacy. This is because ants relocated nests into and/or under these attractive nesting materials, thus increasing their exposure to toxic insecticide residues. Our results demonstrate efficacy of nonrepellent liquid insecticides as indoor treatments for the control of Pharaoh ants and possibly as exterior perimeter treatments. PMID- 15889743 TI - Effects of interspecific competition between two urban ant species, Linepithema humile and Monomorium minimum, on toxic bait performance. AB - We evaluated the effects of interspecific competition on ant bait performance with two urban pest ants, the Argentine ant, Linepithema humile (Mayr), and the little black ant, Monomorium minimum (Buckley). In a laboratory study, the impact of a solid sulfluramid bait on M. minimum was diminished when L. humile were present, whereas the presence of M. minimum reduced the performance of a liquid fipronil bait against L. humile. Argentine ants were not adversely affected by sulfluramid bait at any time, whereas M. minimum was unaffected by fipronil bait until 14 d of exposure. In field studies measuring diel foraging activity, M. minimum seemed to delay L. humile foraging to food stations by approximately 30 min during summer 2001. However, L. humile subsequently recruited to food stations in very high numbers, thereby displacing M. minimum. L. humile visited food stations over an entire 24-h period, whereas M. minimum was only observed visiting food stations during daylight hours. Adjusting the timing of bait placement in the field may minimize any negative effects of interspecific competition between these two species on toxic bait performance. PMID- 15889744 TI - Effect of Summon Preferred Food Source on feeding, tunneling, and bait station discovery by the formosan subterranean termite (Isoptera: Rhinotermitidae). AB - This study evaluated the effect of Summon Preferred Food Source on feeding, tunneling, and bait station discovery by the Formosan subterranean termite, Coptotermes formosanus Shiraki. Bioassays were conducted to determine whether Summon disks affected the aggregation and feeding behavior of termites and to determine whether the presence of Summon disks caused increased recruitment of termites to wood blocks. When termites encountered the disk, they immediately clustered on top of the disk. Termites were observed aggregating on top of the disk throughout the experiment. Consumption of Summon disks was significantly greater than consumption of cardboard disks in paired choice tests. The presence of a Summon disk on top of a wood block caused a significant increase in consumption of the wood block. Bioassays also were conducted to determine whether water extracts of Summon disks affected termite behavior. Consumption of filter paper disks treated with a water extract of Summon disks was significantly greater than consumption of control filter paper disks. Termites tunneled through sand treated with a water extract of Summon disks faster than they tunneled through untreated sand. In a field test, the rate of infestation of monitoring stations with a Summon disk was 3 times greater than the rate of infestations of stations without a disk. PMID- 15889745 TI - Valuation of efficacy and nonrepellency of indoxacarb and fipronil-treated soil at various concentrations and thicknesses against two subterranean termites (Isoptera: Rhinotermitidae). AB - The efficacy and nonrepellency of indoxacarb (150 SC, 150 g [AI]/liter) and fipronil (Termidor SC, 9.1% [Al]) against field-collected eastern subterranean termite, Reticulitermes flavipes (Kollar), and the Formosan subterranean termite, Coptotermes formosanus Shiraki, were evaluated for mortality and penetration into treated soil in laboratory glass tube bioassays. Both insecticides were tested at five concentrations (0, 1, 10, 50, and 100 ppm) and two thicknesses (20 and 50 mm) of treated soil. Indoxacarb caused significantly greater mortality than controls at all treatment thicknesses of > or = 10 ppm, but not at 1 ppm. Concentration and treatment thickness of indoxacarb significantly affected termite mortality. Eastern subterranean termites were significantly more susceptible to indoxacarb than Formosan subterranean termites, but there were no intercolony differences in either species. Termites completely penetrated through all treatment thickness of indoxacarb-treated soil at all concentrations, except one of the six Formosan subterranean termite replicates of 50 mm at 50 ppm, when all termites were killed before tunneling through the treated soil. Fipronil resulted in significantly faster and greater termite mortality than indoxacarb at corresponding concentrations. Concentration and treatment thickness of fipronil also significantly affected termite mortality. There was no intercolony difference in susceptibility to either insecticide in either termite. Both termite species completely penetrated 20-mm treatments of all tested fipronil concentrations, as well as 50-mm soil treated with fipronil at < or = 10 ppm. At 50 and 100 ppm fipronil, termites tunneled only a mean of 87 +/- 0.21 and 47 +/- 0.18% deep into 50-mm treated soil, respectively, before death. Both insecticides demonstrated a delayed mode of activity and nonrepellency against the two termite species. PMID- 15889746 TI - Using a generational time-step model to simulate dynamics of adaptation to transgenic corn and crop rotation by western corn rootworm (Coleoptera: Chrysomelidae). AB - We expanded a simulation model of the population dynamics and genetics of the western corn rootworm for a landscape of corn, soybean, and other crops to study the simultaneous development of resistance to both crop rotation and transgenic corn. Transgenic corn effective against corn rootworm was recently approved in 2003 and may be a very effective new technology for control of western corn rootworm in areas with or without the rotation-resistant variant. In simulations of areas with rotation-resistant populations, planting transgenic corn to only rotated cornfields was a robust strategy to prevent resistance to both traits. In these areas, planting transgenic corn to only continuous fields was not an effective strategy for preventing adaptation to crop rotation or transgenic corn. In areas without rotation-resistant phenotypes, gene expression of the allele for resistance to transgenic corn was the most important factor affecting the development of resistance to transgenic corn. If the allele for resistance to transgenic corn is recessive, resistance can be delayed longer than 15 yr, but if the resistant allele is dominant then resistance usually developed within 15 yr. In a sensitivity analysis, among the parameters investigated, initial allele frequency and density dependence were the two most important factors affecting the evolution of resistance. We compared the results of this simulation model with a more complicated model and results between the two were similar. This indicates that results from a simpler model with a generational time-step can compare favorably with a more complex model with a daily time-step. PMID- 15889747 TI - Analysis of the dynamics of adaptation to transgenic corn and crop rotation by western corn rootworm (Coleoptera: Chrysomelidae) using a daily time-step model. AB - Western corn rootworm, Diabrotica virgifera virgifera LeConte, has overcome crop rotation in several areas of the north central United States. The effectiveness of crop rotation for management of corn rootworm has begun to fail in many areas of the midwestern United States, thus new management strategies need to be developed to control rotation-resistant populations. Transgenic corn, Zea mays L., effective against western corn rootworm, may be the most effective new technology for control of this pest in areas with or without populations adapted to crop rotation. We expanded a simulation model of the population dynamics and genetics of the western corn rootworm for a landscape of corn; soybean, Glycine max (L.); and other crops to study the simultaneous development of resistance to both crop rotation and transgenic corn. Results indicate that planting transgenic corn to first-year cornfields is a robust strategy to prevent resistance to both crop rotation and transgenic corn in areas where rotation-resistant populations are currently a problem or may be a problem in the future. In these areas, planting transgenic corn only in continuous cornfields is not an effective strategy to prevent resistance to either trait. In areas without rotation resistant populations, gene expression of the allele for resistance to transgenic corn, R, is the most important factor affecting the evolution of resistance. If R is recessive, resistance can be delayed longer than 15 yr. If R is dominant, resistance may be difficult to prevent. In a sensitivity analysis, results indicate that density dependence, rotational level in the landscape, and initial allele frequency are the three most important factors affecting the results. PMID- 15889748 TI - Carbaryl resistance in Mexican strains of the southern cattle tick (Acari: Ixodidae). AB - Susceptibility to carbaryl in six Mexican strains of the southern cattle tick, Boophilus microplus (Canestrini), was evaluated with the Food and Agricultural Organization larval packet test. Tick strains from the cattle fever tick quarantine zone in Texas were more susceptible to carbaryl than to coumaphos or diazinon. Compared with the susceptible reference (Gonzalez) strain, Mexican tick strains demonstrated 10.9-59.5-fold resistance to carbaryl. Significant cross resistance was found between carbaryl and the organophosphate acaricides coumaphos and diazinon. Bioassay results with synergists suggested that metabolic detoxification mechanisms did not play a major role in carbaryl resistance. Resistance to carbaryl was likely conferred by insensitive acetylcholinesterase. The implications of carbaryl resistance in tick eradication and control also are discussed. PMID- 15889749 TI - Impact of melon accessions resistant to aphids on the demographic potential of silverleaf whitefly. AB - Bemisia tabaci (Gennadius) and Aphis gossypii Glover are devastating melon, Cucumis melo L., pests. The geographic areas where they occur overlap, and the same chemicals are used to control both of them. Therefore, to reduce pesticide use, it would be necessary to breed melon lines that simultaneously express a resistance to both insects. Female survival; the time when reproduction starts, peaks, and ends; the number of female offspring at the reproductive peak; and total reproduction (S) were determined under semicontrolled conditions for B. tabaci kept in clip-cages on a susceptible melon genotype Vedrantais, and 12 potential resistant accessions, particularly genotypes expressing the Vat gene controlling resistance to A. gossypii. By using the Lewontin triangular reproductive function and Bootstrapping, the intrinsic rate of increase (r) and its variance were calculated. Statistical analysis showed that the parameter S was as relevant as r for discriminating between the melon accessions. Three genotypes were potential genitors of resistance to the whitefly: PI 161375, PI 414723, and PI 532841. Those possessing the Vat gene were either resistant (PI 161375 and PI 414723) or susceptible (Margot, IsoVat R, and AR 5). This demonstrated the ineffectiveness of Vat against B. tabaci. In this article, we propose a strategy to breed lines that express resistance to aphids and whiteflies on the short-term. PMID- 15889750 TI - Detached leaf assay to screen for host plant resistance to Helicoverpa armigera. AB - The noctuid Helicoverpa armigera (Hubner) is a major insect pest of chickpea Cicer arietinum L., pigeonpea Cajanus cajan (L.) Millsp., peanut Arachis hypogaea L., and cotton Gossypium spp., and host plant resistance is an important component for managing this pest in different crops. Because of variations in insect density and staggered flowering of the test material, it is difficult to identify cultivars with stable resistance to H. armigera across seasons and locations. To overcome these problems, we standardized the detached leaf assay to screen for resistance to this pest in chickpea, pigeonpea, peanut, and cotton under uniform insect pressure under laboratory conditions. Terminal branch (three to four fully expanded leaves) of chickpea, first fully expanded leaf of cotton, trifoliate of pigeonpea, or quadrifoliate of peanut, embedded in 3% agar-agar in a plastic cup/jar of appropriate size (250-500-ml capacity) infested with 10-20 neonate larvae can be used to screen for resistance to H. armigera. This technique keeps the leaves in a turgid condition for approximately 1 wk. The experiments can be terminated when the larvae have caused > 80% leaf damage in the susceptible check or when differences in leaf feeding between the resistant and susceptible checks are maximum. Detached leaf assay can be used as a rapid screening technique to evaluate germplasm, segregating breeding materials, and mapping populations for resistance to H. armigera in a short span of time with minimal cost, and under uniform insect infestation. It also provides useful information on antibiosis component of resistance to the target insect pest. PMID- 15889751 TI - Laboratory and field evaluations of transgenic soybean exhibiting high-dose expression of a synthetic Bacillus thuringiensis cry1A gene for control of Lepidoptera. AB - Transgenic lines of soybean, Glycine max (L.) Merrill, expressing a synthetic cry1A gene (tic107) from Bacillus thuringiensis (Bt), were evaluated in screenhouse and conventional field trials for efficacy against lepidopteran pests. In screenhouse trials, Bt soybean and negative checks (isogenic segregants and parental lines) were evaluated against Anticarsia gemmatalis Hubner and Pseudoplusia includens (Walker) in the United States and against A. gemmatalis, Epinotia aporema (Walsingham), Rachiplusia nu (Guenee), and Spilosoma virginica (F.) in Argentina. Bt soybean exhibited virtually complete efficacy against each of these pests, whereas negative checks suffered significant damage. Bt soybean and negative checks also were evaluated in conventional trials against native populations of A. gemmatalis and P. includens in the southeastern United States. Each of these insects caused significant damage to negative checks in one or more locations, whereas Bt soybean exhibited virtually complete efficacy against these pests. In the laboratory, lyophilized leaf tissues from Bt soybean incorporated in artificial diet at a concentration representing a 25-fold dilution of fresh tissue caused complete mortality of A. gemmatalis and near complete mortality of P. includens neonates after 11 d, whereas mortality on negative checks did not exceed 10% for either insect. Average TIC107 expression approached or exceeded 50 microg/g fresh weight at V3 stage of growth and 200 microg/g by R6 stage of growth. These results demonstrate that expression of TIC107 in soybean can not only achieve highly efficacious control of several lepidopterans under field conditions but also provide a high dose for effective insect resistance management. PMID- 15889752 TI - Yield response and categories of resistance to Russian wheat aphid in four Dn4 hard red winter wheat cultivars. AB - A field experiment was conducted to determine whether resistance to Russian wheat aphid, Diuraphis noxia (Mordvilko), conferred by the Dn4 gene is affected by genetic background. This was done by comparing the yield responses to Russian wheat aphid-resistant wheat containing Dn4, derived through the backcross method, to those of the corresponding recurrent parents. Infested resistant cultivars had fewer Russian wheat aphids per tiller than infested susceptible cultivars at the Lamar and Fort Collins, CO sites but not at the Akron, CO site. At the Lamar site, resistant cultivars yielded more than the susceptible cultivars. 'Prairie Red' and 'Yumar' were more resistant than 'Prowers', especially at the higher infestation level. Resistance in these cultivars was categorized in a laboratory experiment to confirm this differential expression of resistance. Resistance in Prairie Red, 'Halt', 'Prowers 99', and Yumar was categorized at three plant growth stages. Antibiosis was expressed as reductions in maximum number of nymphs produced per 24 h and intrinsic rate of increase. The maximum number of nymphs produced per 24 h was reduced in Halt and 'Lamar'. Averaged over cultivars, the intrinsic rate of increase was less at jointing than at the seedling or tillering growth stages. Tolerance was expressed in the resistant cultivars as reduced chlorosis and leaf rolling. Growth reductions in infested Prowers 99 plants was less than the other cultivars. This study confirms that some cultivars containing Dn4 may express antibiosis and tolerance, whereas others may not show the same categories. Thus, expression is affected by genetic background. PMID- 15889753 TI - Molecular mapping of sorghum genes expressing tolerance to damage by greenbug (Homoptera: Aphididae). AB - Genetic linkage maps are fundamental for the localization of genes conferring tolerance to greenbug, Schizaphis graminum (Rondani), feeding damage in sorghum, Sorghum bicolor (L.) Moench. Thirteen linkage groups (LGs) containing 60 simple sequence repeat (SSR) loci were mapped by using a set of sorghum recombinant inbred lines (RILs) obtained from the cross '96-4121' (greenbug-tolerant parent) x Redlan (greenbug-susceptible parent). The LG spanned a distance of 603.5 cM, with the number of loci per LG varying from 2 to 14. Seventeen additional SSR loci were unlinked at a log of odds value of 3.0. Based on chlorophyll loss occurring after greenbug feeding, visual damage ratings, and soil plant analysis development (SPAD), chlorophyll-loss indices were recorded for each RIL and for the parents used in the cross. Composite-interval mapping identified three quantitative trait loci (QTLs) associated with biotype I and five QTLs associated with biotype K. The amount of phenotypic variation explained by these QTLs ranged from 9 to 19.6%. The identification of QTLs that influence greenbug tolerance will not only facilitate the use of marker-assisted selection in sorghum breeding programs but also will provide a solid foundation for detailed characterization of individual loci implicated in greenbug tolerance in sorghum. PMID- 15889754 TI - Resistance to Rhopalosiphum padi (Homoptera: Aphididae) in three triticale accessions. AB - Experiments were conducted to identify and characterize host plant resistance to bird cherry-oat aphid, Rhopalosiphum padi (L.), in various wheat and wheat-grass hybrids. Initial tests screened for resistance to R. padi among 12 grass accessions (eight wheat [Triticum aestivum L.], three triticale [XTriticosecale Wittmack], and 1XElytricum [Elytrigia elongata [Host] Nevski x Triticum aestivum hybrid]). R. padi had less population growth on triticale accessions '8TA5L' (PI 611760) and 'Stniism 3' (PI 386156) than on other accessions, but nymphiposition by R. padi did not differ among the 12 accessions. Follow-up experiments were conducted to characterize antibiosis, antixenosis, and tolerance to R. padi in three wheat and three triticale accessions. In antibiosis experiments, Stniism 3 and triticale 'H7089-52' (PI 611811) prolonged time to reproduction by R. padi compared with that on wheat accessions 'Arapahoe' (PI 518591), 'KS92WGRC24' (PI 574479), and 'MV4' (PI 435095), whereas time to reproduction on 8TA5L was intermediate and did not differ from that on the other five accessions. Also, R. padi produced fewest progeny on Stniism 3, and fewer progeny on 8TA5L than on H7089-52, Arapahoe, KS92WGRC24, and MV4. Stniism 3 showed antixenosis, because fewer winged R. padi selected Stniism 3 than Arapahoe, H7089-52, or MV4 in choice tests. In tolerance experiments, a 300 aphid-day infestation of R. padi limited shoot length of Arapahoe and KS92WGRC24 plants. Shoot lengths did not differ between infested and noninfested seedlings of MV4, 8TA5L, H7089-52, and Stniism 3, indicating tolerance to R. padi in these accessions. Triticale accessions 8TA5L, H7089-52, and Stniism 3 and MV4 wheat may be meaningful sources of R. padi resistance for small-grain breeding programs, and Stniism 3 may be particularly valuable, given reports of its additional resistance to the Russian wheat aphid, Diuraphis noxia (Mordvilko). PMID- 15889755 TI - Whitefly Bemisia tabaci (Homoptera: Aleyrodidae) infestation on cassava genotypes grown at different ecozones in Nigeria. AB - Large-scale screening of cassava, Manihot esculenta Crantz, genotypes for resistance to infestation by whitefly Bemisia tabaci Gennadius, the vector of cassava mosaic geminiviruses, is limited. A range of new cassava elite clones were therefore assessed for the whitefly infestation in the 1999/2000 and 2000/2001 cropping seasons in experimental fields of International Institute of Tropical Agriculture, Ibadan, Nigeria. On each scoring day, between 0600 and 0800 hours when the whiteflies were relatively immobile, adult whitefly populations on the five topmost expanded leaves of cassava cultivars were counted. All through the 6-mo scoring period, there was a highly significant difference in whitefly infestation among the new cassava elite clones. Vector population buildup was observed in Ibadan (forest-savanna transition zone) and Onne (humid forest), 2 mo after planting (MAP). Mean infestation across cassava genotypes was significantly highest (16.6 whiteflies per plant) in Ibadan and lowest in Zaria (0.2). Generally, whitefly infestation was very low in all locations at 5 and 6 MAP. During this period, cassava genotypes 96/1439 and 91/02324 significantly supported higher infestations than other genotypes. Plants of 96/1089A and TMS 30572 supported the lowest whitefly infestation across cassava genotypes in all locations. The preferential whitefly visitation, the differences between locations in relation to whitefly population, cassava mosaic disease, and the fresh root yield of cassava genotypes are discussed. PMID- 15889756 TI - Relationship between supercooling point and mortality at low temperatures in Indianmeal moth (Lepidoptera: Pyralidae). AB - Indianmeal moth, Plodia interpunctella (Hubner), is classified as a freeze intolerant organism and one of the most cold-tolerant stored-product pests. The objective of this study was to determine the relationship between mortality at low temperatures after minimum exposure and the supercooling point (SCP) for laboratory-reared P. interpunctella at different stages of development. This relationship also was analyzed for field-collected, cold-acclimated fifth instars. Mean SCP of laboratory-reared larvae (i.e., feeding stage) was consistently above approximately -16 degrees C. Mean SCP of laboratory-reared pupae and adults (i.e., nonfeeding stages) and field-collected, cold-acclimated fifth instars was consistently below approximately -21 degrees CP seemed to be the boundary between survival and death for larvae. However, it seemed that a 1 min exposure was not sufficient to cause larval mortality at the SCP. Alternatively, for both pupae and adults, the SCP seemed not to play an important role in their survival at low temperatures, with significant mortality observed at temperatures higher than the mean SCP. Adults were the most susceptible to low temperatures with no survival occurring at -20 degrees C, > 3 degrees C above its mean SCP. Results of this investigation demonstrate that P. interpunctella has a different response to low temperatures depending on stage of development and cold acclimation. Classifying P. interpunctella only as a freeze-intolerant organism disregards the occurrence of prefreeze mortality in this species. Therefore, a reclassification of this species (e.g., chill tolerant or chill susceptible) based on the extent of prefreeze mortality and the temperature and time of exposure at which it occurs is suggested. PMID- 15889757 TI - Reproductive performance of Tribolium castaneum (Coleoptera: Tenebrionidae) exposed to the minimum heat treatment temperature as pupae and adults. AB - Managing stored-product insect pests by heating the ambient air of a food processing facility to high temperatures (50-60 degrees C), also referred to as heat treatment, is an effective technology that has been used since the early 1900s. The minimum temperature during heat treatment for effective disinfestation is 50 degrees C. The effect of sublethal exposures to 50 degrees C on the reproductive performance of stored-product insects associated with food processing facilities is unknown. The red flour beetle, Tribolium castaneum (Herbst), is a pest commonly found in food-processing facilities worldwide. The adverse effects on fecundity, egg-to-adult survival, and progeny production of T. castaneum exposed as 1-d-old pupae and 2-wk-old adults to 50 degrees C for 60 and 39 min, respectively, were determined in the laboratory. Pupae and adults exposed for the same time periods at 28 degrees C served as the control treatment. Four possible reciprocal crosses were carried out among adults from the heat-treated (50 degrees C) and control (28 degrees C) treatments. The number of eggs produced during the first 2 wk of adult life, survival of these eggs to adulthood, and adult progeny production after 2 and 8 wk of oviposition in treatments representing all four reciprocal crosses were determined. Fecundity, egg-to-adult survival, and adult progeny production decreased by 17-63, 52-63, and 66-78%, respectively, when males, females, and both males and females were exposed to 50 degrees C. These effects were relatively more pronounced in treatments in which pupae were exposed to the high temperature compared to adults, and in exposed females than in males. The impaired reproductive performance in T. castaneum pupae and adults surviving sublethal exposures to the minimum heat treatment temperature is valuable for understanding population rebound following a heat treatment intervention. PMID- 15889758 TI - Positive ratings of quality of care in Veterans Affairs facilities. PMID- 15889759 TI - Dose adjustment over time of etanercept and infliximab in patients with rheumatoid arthritis. AB - Dose escalation of biologics in patients with rheumatoid arthritis may affect the cost of care. Longitudinal claims data from a large U.S. health plan were analyzed retrospectively. A total of 4,426 health plan members had a medication claim for either etanercept (N = 690; mean age, 48.4 yr; 72% female) or infliximab (N = 424; mean age, 54.3 yr; 73% female) during the selection period. The study revealed that the mean dosage in patients receiving infliximab for rheumatoid arthritis symptoms increased by 29% from first to last dose. The mean weekly dosage for etanercept remained stable. Etanercept was associated with a significantly lower hazard of dose increase relative to infliximab. The estimated annual costs of infliximab and its administration varied by 31%, whereas the estimated costs of etanercept remained stable over time. This increased dose of infliximab may translate into increased costs for a payer over time. PMID- 15889760 TI - Managed care pharmacy 101: learning from the students. PMID- 15889761 TI - Effective coverage and reimbursement strategies for etanercept and infliximab in the treatment of rheumatoid arthritis. AB - Early and aggressive treatment of rheumatoid arthritis (RA) can provide cost savings through enhanced and sustained clinical outcomes. Antitumor necrosis factor-alpha (anti-TNF) agents, such as infliximab and etanercept, provide a superior level of clinical benefit, particularly in patients with moderate-to severe RA. Reimbursement of these agents falls under either a pharmacy or medical benefit, depending on their route of administration. However, inconsistencies in reimbursement strategies across plans potentially restrict clinician and patient access to these safe and effective therapies. Benefits should be designed to ensure that all eligible patients have access to anti-TNF agents, with the understanding that earlier treatment of RA with safe and effective agents provides significant cost savings to future insurers through enhanced and sustained outcomes. PMID- 15889762 TI - Economic effects of aspergillosis management in high-risk patients. AB - Aspergillosis mortality has risen exponentially over the last 2 decades, related primarily to the availability of medical treatments and procedures that prolong the survival of patients with other disorders who are at high risk because of immunologic compromise. In addition to excess mortality, aspergillosis results in increased costs for hospitalization and treatment, including management of adverse events associated with conventional amphotericin B treatment. Compared with amphotericin B, the triazole antifungal voriconazole has excellent efficacy and a favorable side-effect profile. Reliable oral bioavailability of voriconazole may permit switching to an oral formulation in the inpatient setting and offers the possibility of earlier hospital discharge and potentially substantial cost savings. PMID- 15889763 TI - Conf idence intervals P values. PMID- 15889764 TI - Discount Medical cards: leading the way to inexpensive care or potential fraud? AB - As the number of uninsured grow and health care costs continue to rise, discount medical card programs can be expected to become more popular. However, since discount medical card companies and products are not subject to insurance laws, a regulatory loophole has created opportunities for fraudulent cards. PMID- 15889765 TI - The match and the future of family medicine. PMID- 15889766 TI - The ethics of alternative medicine: an alternative standard? PMID- 15889767 TI - Observations on MOC. PMID- 15889768 TI - MOC signals end of the road. PMID- 15889769 TI - Bigger is better. PMID- 15889770 TI - Older is wiser. PMID- 15889771 TI - Is it OK to treat your kids? PMID- 15889772 TI - How to conduct a "welcome to Medicare" visit. PMID- 15889773 TI - Match Day 2005: family medicine gains positions, loses U.S. seniors. PMID- 15889774 TI - 10 steps to HIPAA security compliance. PMID- 15889775 TI - What a medical assistant can do for your practice. PMID- 15889776 TI - Encouraging news about family physician recruitment. PMID- 15889777 TI - Making every minute count: tools to improve office efficiency. PMID- 15889778 TI - Keys to improving your listening skills. PMID- 15889779 TI - Having it all, a little at a time. PMID- 15889780 TI - Framing the threat of resistance to quinolones in pharmacologic terms. PMID- 15889781 TI - Optimizing formulary choices of the fluoroquinolones. PMID- 15889782 TI - Health plan management and the disease burden of lower respiratory tract infections. PMID- 15889783 TI - Cure versus eradication. PMID- 15889784 TI - Antibiotic resistance: implications and strategies for the health plan. PMID- 15889785 TI - [Pregna-D'-pentaranes--progestins and antiprogestins: I. Differentiation of biological functions of steroid hormones]. AB - The synthesis, modification, structure, and biological activity in vivo of the 16alpha,17alpha-cycloalkanoprogesterone (pregna-D'-pentarane) analogues of progesterone are described. A possibility of separation of their biological functions has been demonstrated. A systematic synthesis of a set of uniform compounds that differ in a limited number of alterable parameters was developed. It resulted in an instrument useful for the investigation of pathways and mechanisms by which the steroid hormones fulfill their biological functions and for the probable discovery of new functions masked by the wide effects of native compounds. PMID- 15889786 TI - [Biological activity of cholecystokinin-(30-33) tetrapeptide analogs]. AB - Analogues of the endogenous peptide corresponding to the 30-33 sequence of cholecystokinin (Trp-Met-Asp-Phe-NH2) were synthesized, and their biological activity was studied. It was shown that, in rats, the N-succinylated Nle2 analogue of this tetrapeptide exhibits increased anxiolytic properties in the dark-bright chamber test and an enhanced alcohol intake by both the control animals and the long-time alcohol-dependent animals under the conditions of free choice. Introduction of an isopropyl residue into the C-terminal amide of the Nle2 analogue resulted in the appearance of anxiolytic and antialcohol activity and the ability to increase the morphine analgesic effect in the tail-flick test on rats. The two synthesized analogues retained an affinity to cholecystokinin receptors. PMID- 15889787 TI - [Tertiary structure of myelopeptides. II. Conformational analysis of Phe-Arg-Pro Arg-Ile-Met-Thr-Pro, Val-Val-Tyr-Pro-Asp, and Val-Asp-Pro-Pro]. AB - Theoretical conformational analysis was used to study the spatial structure and conformational properties of myelopeptides, bone marrow peptide mediators. The low-energy conformations of myelopeptides MP-4 (Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro), MP-5 (Val-Val-Tyr-Pro-Asp), and MP-6 (Val-Asp-Pro-Pro) were found; the values of dihedral angles of backbone and side chains of the amino acid residues were determined; and the energies of intra- and interresidual interactions were estimated. PMID- 15889788 TI - [Efficient synthesis of 3'-amino-3'-deoxythymidine derivatives]. AB - An efficient method of reduction of 3'-azido-3'-deoxythymidine and its 5' protected derivatives to 3'-aminothymidine derivatives on a palladium catalyst using ammonium formate as a source of hydrogen was suggested. PMID- 15889789 TI - [Determination of oligonucleotide molecular masses by MS-MALDI]. AB - MALDI mass spectrometry (MS) of 14- to 42-mer homogeneous oligonucleotides and their mixtures was carried out using a Vision 2000 instrument (Thermo BioAnalysis, Finnigan, United States). Conditions for the determination of oligonucleotide molecular masses were optimized by applying various matrices and operation modes. The most reproducible results with minimal uncontrolled decomposition of the oligonucleotides including their apurinization during the MALDI MS registration were obtained using 2,4,6-trihydroxyacetophenone as a matrix instead of 3-hydroxypicolinic acid, usually employed in the mass spectrometry of oligonucleotides. Our approach allows the determination of molecular masses of oligonucleotides obtained by chemical synthesis and the evaluation of their component composition and purity. It was applied to the mass spectrometric analysis of oligonucleotides containing a 3'-(methyl-C-phosphonate) group or a modified 1,N6-ethenodeoxyadenosine unit. PMID- 15889790 TI - [Effect of structural factors on the stability of duplexes formed by oligonucleotide conjugates with minor groove ligands]. AB - The effect of structural factors on the stability of duplexes formed by DNA minor groove binders conjugated with oligonucleotide mono- or diphosphoramidates of the general formula Oligo-MGBm (where Oligo is an oligonucleotide; m = 1 or 2; MGB is -L(Py)2R, L(Py)4R, -L(Im)4R, or -L(Py)4NH(CH2)3CO(Py)4R; Py is a 4-aminopyrrol-2 carboxylic acid residue, L is a gamma-aminobutyric acid or an epsilon aminocaproic acid residue, R = OEt, NH(CH2)6NEt2, or NH(CH2)6N+Me3) was studied by the method of thermal denaturation. The mode of binder interaction with minor groove depends on the conjugate structure; it may be of the parallel head to head type for bisphosphoramidates and of the antiparallel head to tail type for monophosphoramidates of a hair-pin structure. The effects of the duplexes with parallel orientation (bisphosphoramidates, MGB is L(Py)4R, m = 2) and those of the hairpin structure with the antiparallel orientation (monophosphoramidates, MGB is L(Py)4(CH2)3CO(Py)4R, m = 1) on Tm values were close. The influence of the linker (L) and substituent (R) structures upon Tm was more pronounced for monophosphoramidate (MGB is L(Py)nR, m = 1) than for bisphosphoramidate (MGB is L(Py)nR, m = 2). No more than two oligopyrrolcarboxamide residues (either in parallel or antiparallel orientations) can be incorporated into the duplex minor groove. Moreover, it was shown by the example of monophosphoramidates (Oligo L(Py)4R and Oligo-L(Py)4NH(CH2)3CO(Py)4R) that the addition of a second ligand capable of incorporation into the minor groove increased Tm of the corresponding duplex in comparison with the duplex formed by the starting monophosphoramidate. At the same time, the introduction of the ligand incapable of incorporating decreased the Tm value. The mode of interaction of the conjugated ligand with the oligonucleotide duplex is determined by its structure. For example, dipyrrolcarboxamide containing an ethoxy group at the ligand C-end stabilizes the duplex due to the stacking interaction with the terminal A*T pair, whereas tetrapyrrolcarboxamides stabilize the duplex by incorporation into the minor groove. PMID- 15889791 TI - [Optimization of transfection properties of DNA-lysine dendrimer complexes]. AB - We studied the possibility of optimizing the DNA transfection properties of carriers based on lysine dendrimers of the third and the fifth generation, including those containing a chloroacetyl or a lipophilic palmitoyl moiety at C end. The use of lysosome-destroying antibiotic chloroquine and an amphipathic polycationic nonadecapeptide JTS-1 was found to enhance the DNA transfecting properties of the lysine dendrimers. The triple complex including DNA, a lysine dendrimer of the third generation modified with lipophylic moieties of palmitic acid at its C-end, and JTS-1 was shown to be comparable in its transfecting activity to a complex containing Escort, a commercial cationic liposome carrier. PMID- 15889792 TI - [Alternative splicing of pre-mRNA encoding the Musca domestica latrophilin-like protein(LLP): primary structures of four spliced forms of mRNA and their protein products]. AB - An internal DNA fragment (approximately 2000 bp) homologous to the conserved regions of genes encoding latrophilin-like proteins (LLPs) was obtained by the PCR technique using degenerate primers to these gene regions. The gene-specific primers were synthesized based on the results of sequencing of the isolated fragment, and all overlapping cDNA fragments of the llp gene encoding the Musca domestica LLP were obtained by the rapid amplification of cDNA 5'- and 3'-ends (5'- and 3'-RACE). Four alternatively spliced mRNAs were found while sequencing the obtained cDNA fragments. Two long mRNAs (approximately 6000 nt) differ in the structures of both the sites encoding signal peptides and 5'-terminal untranslated regions. They encode large proteins (approximately 1800 aa), whose domain organization is similar to that of mammalian latrophilins. Each deduced protein contains a domain with seven transmembrane regions followed by an extended cytoplasmic C-terminal domain. Two other mRNA forms are derived from these long mRNAs; they encode proteins severly truncated at their C-termini (approximately 900 aa). They are composed of only three transmembrane regions and a short unique cytoplasmic C-terminal domain (23 aa). The limitations and drawbacks of the existing 3'-RACE techniques found during study of the long alternatively spliced cDNAs are analyzed, and ways for overcoming these difficulties are proposed. PMID- 15889793 TI - [A method for the preparation of normalized cDNA libraries enriched with full length sequences]. AB - We developed a new method for the preparation of normalized cDNA libraries enriched with full-length sequences. It is based on the properties of the recently characterized duplex-specific nuclease from the hepatopancreas of the Kamchatka crab. The duplex-specific nuclease is thermostable, it effectively cleaves double-stranded DNA and is inactive toward single-stranded DNA (Shagin et al., Genome Res., 2002, vol. 12, pp. 1935-1942). Our method enables the normalization of cDNA samples enriched with full-length sequences without use of laborious and ineffective stages of physical separation. The efficiency of the method was demonstrated in model experiments using cDNA samples from several human tissues. PMID- 15889794 TI - [The NOLA2 and RPS3A genes as highly informative markers for human squamous cell lung cancer]. AB - cDNA libraries enriched with sequences that are differentially transcribed in normal and tumor tissues were prepared using the subtractive hybridization of mixtures of cDNAs from ten patients with squamous cell lung cancer and the corresponding mixtures of cDNAs from normal tissues of the same patients. An analysis of the libraries revealed two genes, NOLA2 and RPS3A, whose expression in patients with squamous cell lung cancer increased by 70%. A high frequency of enhanced expression of these genes in the cancer makes them highly informative markers of squamous cell lung cancer, which, together with other markers, can be used for reliable diagnosis of the disease. PMID- 15889795 TI - [New syntheses of alpha-methyl- and alpha,alpha'-dimethylspermine]. AB - alpha-Methylspermine and alpha,alpha'-dimethylspermine were synthesized in high overall yields starting from N-(benzyloxycarbonyl)-3-aminobutanol in order to study polyamine biochemistry in vitro and in vivo. PMID- 15889796 TI - [New oxaanalogues of spermine]. AB - A new isosteric charge-deficient spermine analogue, 1,12-diamino-4,9-diaza-5 oxadodecan, and O-(7-amino-4-azaheptyl)oxime of 3-aminopropanal, a stable analogue of the Schiff base intermediate in the enzymatic oxidation of spermine, were synthesized. The possible use of these compounds for the inhibition of spermine oxidase is discussed. PMID- 15889797 TI - [Universal method for single nucleotide substitution identification]. AB - A new approach to the identification of point mutations by allele-specific PCR was proposed. The mutation R408W of the human phenylalanine hydroxylase gene was used as a model. A high specificity of the approach was achieved by the use of primers partially complementary to the genomic DNA. Polyethylene glycol covalently attached to one of the allele-specific primers provides for the differential identification of the PCR products due to a change in electrophoretic mobility. PMID- 15889798 TI - The power of partnering. PMID- 15889799 TI - Extreme nephrology nursing. PMID- 15889800 TI - The value of evidence-based practice in nephrology nursing. PMID- 15889801 TI - Water treatment for hemodialysis--updated to include the latest AAMI standards for dialysate (RD52: 2004) continuing. AB - While nurses may not routinely service the water treatment system or mix the dialysate, they are responsible for understanding all the clinical ramifications of water and dialysate for HD and helping to piece together the entire treatment picture. Although historically the water treatment system has been in the technicians' domain, knowing the technical aspects is important in order for the entire team to work together toward the patients' ultimate well being. This article describes the composition of water treatment systems for hemodialysis as well as the monitoring and testing necessary to assure that both water and dialysate are safe for patient use. PMID- 15889802 TI - Heart rate variability and mortality in patients with end stage renal disease. AB - Heart rate variability (HRV), a measure of autonomic function, is associated with mortality in non-end stage renal disease (ESRD). The purpose of this pilot study was to determine if HRV was predictive of mortality in patients on dialysis and to identify at-risk factors. Patients on chronic hemodialysis (n = 53) were assessed at baseline and again 24 months later. Baseline measures quantified 24 hour HRV, health, depression, and quality of life (QoL). Twenty-four-month data determined mortality. Participants were African American, 49% male, aged 47.8+/ 13.3 years, with 62.4+/-60 months of dialysis. Outcomes of 24-hour HRV measures were impaired for all groups. Factors including exercise and smoking were associated with diminished HRV. The low frequency-high frequency ratio was found to be the most influential HRV determinant of death. The ability to identify patients at-risk for death and to prescribe therapy to reduce risk could have significance for the care of patients with ESRD. PMID- 15889803 TI - Facilitating sleep for patients with end stage renal disease. AB - Patients with end-stage renal disease (ESRD) are confronted with many changes and problems; a lack of sleep may be one of the most prevailing difficulties they endure. Poor sleep is often the result of restless leg syndrome (RLS), periodic leg movements during sleep (PLMS), sleep apnea syndrome (SAS), or depression. Nurses who provide health care to these patients can use timely interventions to assist the patients to achieve better sleep. PMID- 15889804 TI - The benefits of IV iron therapy in treating anemia in patients with renal disease and comorbid cardiovascular disease. AB - It is important for nephrology nurses to understand the relationship that exists between renal disease, cardiac disease, and anemia. Even mild cases of chronic kidney disease (CKD) have been associated with an increase in adverse cardiovascular outcomes. And anemia, which can result from both CKD and congestive heart failure, has been shown to exacerbate the adverse consequences of these conditions. An early, aggressive correction of anemia in patients with CKD can be implemented to break this cycle and stop disease progression. Studies have shown that anemia correction improves both cardiac and renal function and can result in increased hemoglobin levels, decreased number of hospital days, and improved quality of life. An effective strategy for managing anemia in patients with renal disease and comorbid cardiovascular disease includes the administration of both recombinant human erythropoietin and intravenous iron. In addition, the nephrology nurse plays an integral role in managing anemia and improving outcomes in these patients. Therefore, the nephrology nurse should have an increased awareness of the link between anemia and renal/cardiac disease as well as available treatment options. PMID- 15889805 TI - Patient education and the nursing process: meeting the patient's needs. AB - The ultimate goal of patient educational programs is to achieve long-lasting changes in behavior by providing patients with the knowledge to allow them to make autonomous decisions to take ownership of their care as much as possible and improve their own outcomes. Patient education is an integral part of the nursing process, and nephrology nurses can use this process to assess, plan, implement, and evaluate an effective and individualized patient education program. PMID- 15889806 TI - Initiation of dialysis in a patient with diabetes and chronic kidney disease. PMID- 15889807 TI - National vascular access improvement initiative: "Fistula First". PMID- 15889808 TI - Retrograde arterial needle placement improves dialysis adequacy. PMID- 15889809 TI - The arterial needle can be placed in the direction of flow to achieve effective treatment. PMID- 15889810 TI - Let's empower patients with the choice of self-cannulation! PMID- 15889811 TI - Needle placement is paramount to achieving effective dialysis and preserving vascular accesses. PMID- 15889812 TI - The role of cinacalcet in treating secondary hyperparathyroidism. AB - Compelling evidence exists to use cinacalcet as a primary therapy for the treatment of secondary HPT in patients requiring dialysis. The benefits of directly lowering PTH in combination with improving calcium-phosphorus homeostasis provide a means to better achieve the K/DOQI bone guidelines. Cinacalcet is usually well tolerated and safe to use, making it possible to treat a wide range of patients with varying levels of disease severity. The ideal course of action for this novel therapy is for nephrology nurses and renal dietitians to use a team approach in the management of secondary HPT. PMID- 15889813 TI - Home dialysis, home dialysis central, and what you can do today. PMID- 15889814 TI - Caring for patients with ESRD in medical-surgical areas. PMID- 15889815 TI - [Characteristics of the brain support of verbal processes in children with difficulties in writing and reading]. PMID- 15889816 TI - [Brain mechanisms of speech process in stuttering]. PMID- 15889817 TI - [Subcortical and limbic structures and P300 in patients with schizophrenia]. PMID- 15889818 TI - [Sensorimotor and cognitive laterality profiles]. PMID- 15889819 TI - [Characteristics of interregional interactions of cortical fields at different stages of normal and hypnotic sleep (according to EEG data)]. PMID- 15889820 TI - [Effect of musical accompaniment on the operation performance of subjects with different anxiety levels]. PMID- 15889821 TI - [Effects of rotating shift work on central hemodynamic parameters and the constant potential of the brain]. PMID- 15889822 TI - [Parameters of thermohomeostasis and psychophysiological monitoring in the evaluation of the phasic structure of adaptation during 240-day isolation]. PMID- 15889823 TI - [Change in the power of EEG activity in the alpha range in response to tonic pain stimulation of the distal joint of the little finger]. PMID- 15889824 TI - [Dynamics of the states of heart rate regulation during 24-h monitoring]. PMID- 15889825 TI - [Characteristics of the cardiointervalogram in newborns during sleep]. PMID- 15889826 TI - [Low-volume discrete plasmapheresis in therapy of acute destructive lung and pleural diseases]. PMID- 15889827 TI - [Relationship between hormonal activity and trophoblastic beta1 glycoprotein in pregnancy]. PMID- 15889828 TI - [Age-related changes in lipid peroxidation in various regions of the central nervous system]. PMID- 15889829 TI - [A view on the cardiac rhythm control: intracardial regulation]. PMID- 15889830 TI - [An apparatus for mechanical ventilation of the lungs with the possibility of simultaneous control by a subject of the air supply rate and the inspiratory volume]. PMID- 15889831 TI - [Increased antibody titers to bifidobacteria as a marker of intestinal dysbacteriosis]. PMID- 15889832 TI - [Fluctuations of the antianxiety effect of valerian and grandaxin during daytime wakefulness in humans with different chronotypes]. PMID- 15889833 TI - [Lability of regulatory systems as a component of mobilizational readiness in sports]. PMID- 15889834 TI - [Characteristics of the physical state of schoolchildren before and after sanatorium rehabilitation]. PMID- 15889835 TI - Elective abdominal delivery--should mothers have the right to choose? PMID- 15889836 TI - Mbewu ducks AIDS deaths. PMID- 15889837 TI - Criteria for chronic dialysis. PMID- 15889838 TI - Caesarean section--claims and concerns. PMID- 15889839 TI - Blood on the racial walls. PMID- 15889840 TI - The insanity of a criminal justice system. PMID- 15889841 TI - As sweet by any other name? PMID- 15889842 TI - Lawmakers 'protected pharmacies'. PMID- 15889843 TI - Impact of the HIV/AIDS pandemic on non-communicable disease prevention. PMID- 15889844 TI - Hormone replacement therapy--differing guidelines. PMID- 15889845 TI - AIDS prevention through peer education. PMID- 15889846 TI - Dramatic decline in abortion mortality due to the Choice on Termination of Pregnancy Act. PMID- 15889847 TI - Telemetric antenatal fetal monitoring. PMID- 15889848 TI - Nevirapine toxicity--implications for management of South African patients. PMID- 15889849 TI - Private-sector caesarean sections in perspective. PMID- 15889850 TI - Evaluation of an HIV/AIDS peer education programme in a South African workplace. AB - OBJECTIVES: To evaluate a South African workplace HIV/AIDS peer-education programme running since 1997. METHODS: In 2001 a cross-sectional study was done of 900 retail-section employees in three geographical areas. The study measured HIV/AIDS knowledge, attitudes towards people living with HIV/AIDS, belief about self-risk of infection, and condom use as a practice indicator. The impact of an HIV/AIDS peer-education programme on these outcomes was examined. RESULTS: Training by peer educators had no significant impact on any outcome. Fifty-nine per cent of subjects had a good knowledge score, 62% had a positive attitude towards people with HIV/AIDS, 34% used condoms frequently, and the majority of participants (73%) believed they were at low risk of infection. Logistical regression showed that a very small proportion of the variance in the four outcomes was explained by potential determinants of interest (8% for knowledge, 6% for attitude, 7% for risk and 17% for condom use). CONCLUSIONS: The HIV peer education programme was found to be ineffective and may have involved an opportunity cost. The programme contrasts with more costly comprehensive care that includes antiretrovirals. The private sector appears to have been as tardy as the public sector in addressing the epidemic effectively. PMID- 15889851 TI - How well does South Africa's National Health Act regulate research involving children? AB - Currently there are no laws in South Africa regulating the rights of research participants. The National Health Act is the first attempt by the legislature to use the law to protect research participants, including children. This article describes the strengths and limitations of the provisions, implications for researchers and research ethics committees, and makes recommendations. Strengths of the Section include that it enables the Minister of Health to issue regulations detailing protections for research participants, it supplements existing law on consent, it introduces the concept of the 'best interests' of the child and it creates procedural safeguards. Limitations of the Section include that it does not set an independent age for consent to research, it focuses on informed consent and not other protections, it is inconsistent with existing or draft legislation and ethical guidelines, and it retains the contested distinction between 'therapeutic' and 'non-therapeutic' research. Poor drafting and inconsistencies also impede interpretation. The implications for researchers are that it facilitates so-called 'non-therapeutic' research on children. However, procedural burdens for obtaining consent are created. Research Ethics Committees (RECs) will have to work with the 'therapeutic' and 'non-therapeutic' distinction as well as new concepts such as 'best interests' of the child, and ensure that consent procedures comply with the Act. We conclude that while the Act is an important development in the law, it is flawed in places. We recommend that amendments be made and that capacity development be provided to stakeholders. PMID- 15889852 TI - The South African Medical Research Council's Guidelines on Ethics for Medical Research--implications for HIV-preventive vaccine trials with children. AB - Children are at risk of HIV infection, stand to benefit from the development of HIV preventive vaccines, and therefore should be enrolled in trials of HIV vaccines in order to generate relevant safety, immunogenicity and efficacy data. In South Africa, the national vaccine initiative is considering the future conduct of trials involving children; this requires an analysis of the current ethical framework, including elements that facilitate or constrain the conduct of such trials. In this article, we examine the Medical Research Council (MRC)'s Guidelines on Ethics for Medical Research: General Principles (Book 1), and their provisions on research involving children. We argue that this set of influential guidelines includes provisions on research with children that are conceptually problematic and may prohibit critical research with healthy (but at-risk) child participants, including trials of HIV-preventive vaccines. We recommend that Book 1 provisions should be redrafted to reflect a balance between protecting children from research-related risks and testing interventions critical to their health. PMID- 15889853 TI - Neoteric medicine revisited. PMID- 15889854 TI - Prescription for AEDs? PMID- 15889855 TI - The CAM controversy: should Tennessee embrace alternative medicine? PMID- 15889856 TI - State action doctrine: restoring the balance. PMID- 15889857 TI - Loss prevention case of the month. Acting within your own limits. PMID- 15889858 TI - Problem gambling. PMID- 15889859 TI - OIG opinions on malpractice subsidies and gainsharing: a legal analysis. PMID- 15889860 TI - Post-traumatic thoracic arteriovenous fistulas. AB - Post-traumatic arteriovenous fistulas (AVF) between vascular structures in the thorax rarely occur. We present an unusual case of the delayed diagnosis of two post-traumatic thoracic AVF in a 70-year-old male. He had abnormal communications between an intercostal artery and an intercostal vein as well as an intercostal artery and a pulmonary vein. He has remained asymptomatic and has not developed a complication from these abnormal vascular structures in a three-year follow up. PMID- 15889861 TI - Emergency department evaluation of febrile children after the introduction of Prevnar. AB - The Emergency Department work-up of febrile children is largely driven by the risk of occult bacteremia. This study was designed to determine if emergency medicine doctors had changed their work-up of febrile children after introduction of the pneumococcal conjugate vaccine (PCV) in 2000. We surveyed 411 licensed emergency doctors in Tennessee in 2001. Participants were presented with a hypothetical eight-month-old, well-appearing child with a temperature of 102.2 degrees F with no source of infection. They were asked about practice setting, years in practice, laboratory evaluation and whether their work-up of febrile children had changed in the past year. Of those surveyed, 238 (58%) of 411 completed a survey. Of these, 39 were excluded, leaving a study group of 199. Thirty-two (16%) of 196 respondents to the practice-setting question worked in university-affiliated hospitals, and 164 (84%) worked in community hospitals. Twenty-seven (14%) of 196 respondents had been in practice for five years or less, and 169 (86%) respondents had been in practice for greater than five years. One-hundred-and-thirty-eight (69%) of 199 respondents chose to order a complete blood count and 92 (46%) respondents ordered blood cultures. Overall, 22 (11%) respondents stated that they had changed their work-up in the past year. This survey of emergency doctors demonstrates that changes in the work-up of the febrile child were beginning to occur in the year after the introduction of PCV. Because of the dramatic decrease in invasive pneumococcal disease since introduction of the vaccine, future surveys will be needed to determine if the evaluation of febrile children has changed since this survey was conducted. PMID- 15889862 TI - Tennessee trauma care system plan, Part II. AB - Tennessee implemented a statewide trauma care system in 1988. This system serves the state of Tennessee and supports eight neighboring states. The demographics and geography of Tennessee have ensured that nearly all residents have rapid access to the trauma care system. However, since 1988, many changes have occurred in healthcare in general, and trauma care in particular, that point out problems and issues with the Tennessee trauma care system. Therefore, the Tennessee Trauma Care Advisory Council has developed this Trauma Care System Plan to look at needs and opportunities for the future of trauma care in Tennessee. This plan will be presented in four segments: History, Administrative Components, Operational Components, and Clinical Components. PMID- 15889863 TI - The impact of change: child custody and support today. PMID- 15889864 TI - The alcohol 'epidemic': what can be done? PMID- 15889865 TI - Is there a future for academic medicine in the UK? PMID- 15889866 TI - Evidence for inpatient rehabilitation as an effective intervention. AB - Rehabilitation in inpatient settings is expensive and staff-intensive. It is necessary for such services to demonstrate effectiveness to justify this. In contrast to popular notions, evidence for the effectiveness of inpatient rehabilitation does exist and is reviewed in this article. In particular, there is very good evidence for specialized inpatient stroke care and rehabilitation. PMID- 15889867 TI - Choosing appropriate measures in inpatient rehabilitation. AB - This article aims to assist inpatient rehabilitation clinicians to choose appropriate measures. Emphasis is given to measurement processes and interpretation. The authors provide examples of measures commonly reported for five rehabilitation diagnostic groups: stroke, traumatic brain injury, spinal cord injury, multiple sclerosis and cardio-respiratory rehabilitation. PMID- 15889868 TI - A new classification for outcomes in illness and injury. AB - Illness and injury can lead to complex problems for patients and clinicians. A new approach from the World Health Organization (WHO), the International Classification of Functioning, Disability and Health (ICF), may help manage such patients, and evaluate the success of the interventions used. The authors discuss the application of the ICF model. PMID- 15889869 TI - Stem cells as future therapy in cardiology. AB - This article focuses on the key studies relevant to the clinical application of stem-cell research in cardiovascular disease. The authors also discuss current and future directions in clinical cardiovascular stem-cell research, including the potential problems and pitfalls that must be addressed to ensure the safety, as well as the efficacy, of treatment regimens in this rapidly evolving therapeutic field. PMID- 15889870 TI - Aesthetic and functional rhinoplasty. AB - The authors stress that conservative correction should be the main goal in aesthetic and functional rhinoplasty. The surgeon controls the operative event and must be skilled at manipulating and controlling the dynamics of postoperative healing to achieve long-term functional and aesthetic results. PMID- 15889871 TI - The death of Nelson at Trafalgar. PMID- 15889872 TI - Cardiac valvulopathy. PMID- 15889873 TI - Management problems of spontaneous ICH. AB - The management of spontaneous intracerebral haemorrhage (ICH) can be challenging for hospital doctors. Although the management of ICH is covered in stroke guidelines, many difficult clinical questions remain. In this article the authors suggest approaches to ten common and difficult questions. PMID- 15889874 TI - Transforming learning: the challenge of interprofessional education. AB - Interprofessional education (IPE) is the foundation for collaborative practice. It is resource intensive because it requires a shift away from didactic teaching towards a model of facilitated small group learning. This article discusses the ways in which IPE is supported by educational theory and summarizes the increasing evidence for its effectiveness in transforming health-care organizations, leading to increased staff motivation and direct improvements in patient care. PMID- 15889875 TI - Benefits of interprofessional learning: an interprofessional MSc in child health. AB - This article addresses interprofessional education (IPE) using a case study evaluating a multidisciplinary MSc course in child health. The participants felt that the nature of the course increased their interprofessional working skills and professional confidence. They described benefits, including new insights, a balanced variety of views, development of respect and equality between professionals, improved communication and a holistic approach to child health. PMID- 15889876 TI - Forward thinking in orthopaedic surgery prophylaxis. PMID- 15889877 TI - Prostate cancer metastasizing to bone with normal PSA level: a lesson to learn. PMID- 15889878 TI - Intermittent gastric volvulus secondary to gall bladder adhesions: a rare cause of gastric outlet obstruction. PMID- 15889879 TI - A new approach in the treatment of faecaloma of the colon. PMID- 15889880 TI - The 'wandering' abdominal lump: intussusception up to splenic flexure of an ileocaecal adenocarcinoma. PMID- 15889881 TI - Emergency anaesthetic strategies for the bleeding upper airway. PMID- 15889882 TI - In the public's view... Terri Schiavo's 15-year death. PMID- 15889883 TI - End-of-life decisions can be complex, even when patients have a DNR. AB - Proactive communication with your patient and family members can clarify the decision-making process. Even if patient has a do-not-resuscitate (DNR) order, reconfirm patient's wishes when possible. Let family know if you do not believe a treatment will be efficacious. Initiate discussions with patients whom you think should consider having a DNR. PMID- 15889884 TI - Manufacturing techniques help turn ED around. AB - Borrowing quality tools from other industries can help give you a new perspective on the operations of your ED. Computer simulations can help you visualize impact of patient arrivals at peak hours. A flexible approach opens up new options for using existing space more efficiently. 'Manufacturing' approach enables you to break down large processes into more manageable subsets. PMID- 15889885 TI - Will staff restructuring improve psych diagnoses? AB - Shift philosophy and supplement your staff's expertise with specialists to improve psychiatric diagnosis rates. Broaden your trauma-based model to incorporate non-urgent mental health conditions. Increase access to substance abuse specialists, psychologists/psychiatrists, and social workers. Seek to have other departments bear financial burden of making these specialists available to the ED. PMID- 15889887 TI - Simulation competency course is a first. PMID- 15889886 TI - Training with mannequins improves safety, efficiency. AB - Mannequin training offers viable alternative to more standard training approaches. Staff has ability to respond to the same challenging scenarios over and over again. Simulation can be used for pure teaching or for competency examinations. Training also can be used to monitor compliance with patient safety accreditation standards. PMID- 15889888 TI - What are follow-up requirements for on-call physicians? PMID- 15889889 TI - Dissociation of ligand-receptor complexes using magnetic tweezers. AB - We present a new tool for measuring ligand-receptor complex bonds at the single molecule level using magnetic tweezers. Our apparatus allows massively parallel (100-1000) measurements on many single complexes perturbed by constant forces. Compared to other single-molecule techniques, our method is simple, inexpensive, robust, and widely compatible with other techniques. We immobilized specific receptor molecules on the surface of superparamagnetic beads and corresponding ligand molecules on a fixed surface. The beads were allowed to contact the surface so that ligand-receptor binding occurred. A permanent magnet then generated a constant force that pulled the receptors away from the ligands. The rates at which bound species separated at various forces allowed us to characterize the potential energy landscape of the bond and extrapolate bulk solution kinetic rates and transition-state distances. These values agreed with those obtained using bulk and single-molecule methods. PMID- 15889890 TI - Secondary reactions and strategies to improve quantitative protein footprinting. AB - Hydroxyl radical-mediated footprinting permits detailed examination of structure and dynamic processes of proteins and large biological assemblies, as changes in the rate of reaction of radicals with target peptides are governed by changes in the solvent accessibility of the side-chain probe residues. The precise and accurate determination of peptide reaction rates is essential to successfully probing protein structure using footprinting. In this study, we specifically examine the magnitude and mechanisms of secondary oxidation occurring after radiolytic exposure and prior to mass spectrometric analysis. Secondary oxidation results from hydrogen peroxide and other oxidative species generated during radiolysis, significantly impacting the oxidation of Met and Cys but not aromatic or other reactive residues. Secondary oxidation of Met with formation of sulfoxide degrades data reproducibility and inflates the perceived solvent accessibility of Met-containing peptides. It can be suppressed by adding trace amounts of catalase or millimolar Met-NH2 (or Met-OH) buffer immediately after irradiation; this leads to greatly improved adherence to first-order kinetics and more precise observed oxidation rates. The strategy is shown to suppress secondary oxidation in model peptides and improve data quality in examining the reactivity of peptides within the Arp2/3 protein complex. Cysteine is also subject to secondary oxidation generating disulfide as the principal product. The disulfides can be reduced before mass spectrometric analysis by reducing agents such as TCEP, while methionine sulfoxide is refractory to reduction by this reagent under typical reducing conditions. PMID- 15889891 TI - Development of a high-throughput format for solid-phase extraction of enantiomers using an immunosorbent in 384-well plates. AB - An antibody-based solid-phase extraction method for filtered 384-well plates was developed for a medical drug candidate having two enantiomeric forms in order to demonstrate the potential of the use of recombinant antibody fragments as specific and efficient immunosorbents. An immobilization method using a six histidine tag of the antibody fragment and mild oxidation was applied in order to immobilize antibody fragments in an oriented and kinetically stable way that ensured high capacity of the antibody support. Phosphate buffer or plasma spiked with enantiomers were used as samples. Selective solid-phase extraction was followed by liquid chromatography-mass spectrometry analysis. Average recoveries for buffer and plasma samples ranged from 79 to 122% and 80 to 108%, respectively. Good linearity was observed in the concentration range of 30-3000 ng/mL of the enantiomer. PMID- 15889892 TI - Determination of air-to-water partition coefficients using automated multiple headspace extractions. AB - Air-to-water partition coefficients are experimentally determined using a multiple headspace extraction procedure and an automated headspace cell. The approach is first validated with 2-butanone and then applied to a homologous series of methyl ketones. As adsorptions of the most hydrophobic compounds occurred in the sampling cell, technical improvements have been tested. This study represents the first attempt to overcome analyte adsorptions by studying and minimizing the effect of the cell's adsorption of hydrophobic analytes on the determination of their partition coefficients. The present method allows the measurement of several analytes at the same time, in the ppm range, without calibration, and with a limited manpower. PMID- 15889893 TI - Automated multiple headspace extraction procedure: adsorption modeling and determination of air-to-water partition coefficients. AB - The present work makes the first attempt to take into account adsorptions in the determination of partition coefficients by modeling the multiple headspace extraction (MHE) process. Modeling a six-step MHE procedure of a homologous series of methyl ketones revealed that their adsorption-desorption on the walls was the rate-limiting step. Moreover, a comparison between experimental and predicted MHE plots shows that only the last MHE points were affected by adsorption phenomena. Using cell materials with the lowest sorptive properties, partition coefficients were then accurately calculated from the first four MHE steps. This kinetic approach supports previous work in which adsorptions were lowered owing to the choice of sampling cell materials. It also justifies some reproducibility limitations of the MHE quantification procedure mentioned in the literature. PMID- 15889894 TI - Nonspecific protein-carbohydrate complexes produced by nanoelectrospray ionization. Factors influencing their formation and stability. AB - Factors influencing the formation of nonspecific protein-carbohydrate complexes during the nanoelectrospray (nanoES) process have been investigated. Protonated and deprotonated nonspecific complexes of ubiquitin (Ubq) and protonated complexes of carbonic anhydrase (CA) with carbohydrates, ranging in size from mono- to tetrasaccharide, were produced by nanoES and detected with a Fourier transform ion cyclotron resonance mass spectrometer. Both the fraction of protein engaged in nonspecific binding with the carbohydrates and the number of carbohydrates bound to the protein increase with increasing carbohydrate concentration. At a given concentration of protein and carbohydrate, nonspecific binding is favored for small (mono- and disaccharide) or hydrophilic carbohydrates over larger or more hydrophobic molecules, which tend to form gaseous monomer or cluster ions by nanoES. However, the extent of nonspecific binding is insensitive to the structure of the protein, with similar distributions of nonspecific complexes observed for both CA and Ubq. Nonspecific association is also insensitive to the charge state of the complex. A comparable degree of binding is observed for complexes in their protonated and deprotonated forms. Furthermore, the number of bound ligands can exceed significantly the charge state of the complex. Thermal dissociation experiments performed on the gaseous nonspecific complexes reveal that their kinetic stability is sensitive to both the structure of the carbohydrate (i.e., mono- < di- < tri- < tetrasaccharide) and the protein (Ubq < CA) and to the charge state, although no simple relationship between stability and charge state was identified. Taken together, the results of this study suggest that neutral protein-carbohydrate interactions (e.g., hydrogen bonds) contribute significantly and, perhaps, predominantly to the formation and stabilization of the nonspecific complexes. A strategy to minimize the formation of the nonspecific complexes, which is based on the enhancement of gaseous carbohydrate ion formation through the addition of metal salts (e.g., CaCl2) to the nanoES solution, is demonstrated. PMID- 15889895 TI - Multipole-storage-assisted dissociation for the characterization of large proteins and simple protein mixtures by ESI-FTICR-MS. AB - In Fourier transform ion cyclotron resonance mass spectrometry, collisionally activated dissociation (CAD) typically is accomplished within the analyzer ion cell. An alternative approach of multipole-storage-assisted dissociation (MSAD) has previously been demonstrated by inducing collisional fragmentation in the external multipole that is usually employed for ion accumulation. To explore the utility of MSAD for interrogating intact proteins and simple protein mixtures in a multiplexed manner, we have investigated the means of controlling the collisional energy and the fragmentation pattern for this experimental approach. With protein samples in the low micromolar concentration range, the two major experimental parameters affecting MSAD in the hexapole region were found to be the dc offset voltage and accumulation time. While low-energy MSAD of intact proteins yields fragment ions similar to sustained off resonance irradiation collision-activated dissociation (SORI-CAD), high-energy MSAD induces sequential fragmentation for intact proteins to yield a rich variety of singly charged ions in the m/z 600-1200 Da region. Each of the seven proteins (Mr range of 8.5-116 kDa) examined in this study exhibited their own characteristic MSAD fragmentation pattern, which could be used as a signature of the presence of a given protein, even in a mixture. In addition, any MSAD fragment can be isolated and dissociated further by SORI-CAD in an MS3-type experiment inside the FTICR analyzer cell. This presents a novel way to interrogate the identities of these fragment ions as well as obtain amino acid sequence tag information that can be used to identify proteins from mixtures. PMID- 15889896 TI - Liquid chromatography-tandem mass spectrometry determination of nonionic organophosphorus flame retardants and plasticizers in wastewater samples. AB - A comprehensive method for the determination of nonionic organophosphorus flame retardants/plasticizers in wastewater samples by solid-phase extraction (SPE) with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC ESI-MS/MS) is presented. It allows the determination of 11 organophosphorus compounds, trialkyl and trichloralkyl phosphates, triaryl phosphate, and biphosphates together with triphenylphosphine oxide (TPPO). Limits of quantification after SPE of 100 mL of water are between 3 and 80 ng/L, which are adequate for most aqueous samples. The sensitivity of the LC-ESI-MS approach allows direct injection of aqueous sample without preceding extraction for concentrations in the low microg/L range. The method was finally applied to municipal wastewater samples, showing the occurrence of six phosphoric acid triesters and TPPO in both raw and treated municipal wastewaters. PMID- 15889897 TI - Automated 20 kpsi RPLC-MS and MS/MS with chromatographic peak capacities of 1000 1500 and capabilities in proteomics and metabolomics. AB - Proteomics analysis based-on reversed-phase liquid chromatography (RPLC) is widely practiced; however, variations providing cutting-edge RPLC performance have generally not been adopted even though their benefits are well established. Here, we describe an automated format 20 kpsi RPLC system for proteomics and metabolomics that includes on-line coupling of micro-solid phase extraction for sample loading and allows electrospray ionization emitters to be readily replaced. The system uses 50 microm i.d. x 40-200 cm fused-silica capillaries packed with 1.4-3-microm porous C18-bonded silica particles to obtain chromatographic peak capacities of 1000-1500 for complex peptide and metabolite mixtures. This separation quality provided high-confidence identifications of >12 000 different tryptic peptides from >2000 distinct Shewanella oneidensis proteins (approximately 40% of the proteins predicted for the S. oneidensis proteome) in a single 12-h ion trap tandem mass spectrometry (MS/MS) analysis. The protein identification reproducibility approached 90% between replicate experiments. The average protein MS/MS identification rate exceeded 10 proteins/min, and 1207 proteins were identified in 120 min through assignment of 5944 different peptides. The proteomic analysis dynamic range of the 20 kpsi RPLC-ion trap MS/MS was approximately 10(6) based on analyses of a human blood plasma sample, for which 835 distinct proteins were identified with high confidence in a single 12-h run. A single run of the 20 kpsi RPLC-accurate mass MS detected >5000 different compounds from a metabolomics sample. PMID- 15889898 TI - Determination of ephedrine alkaloids in dietary supplement standard reference materials. AB - A suite of five ephedra-containing dietary supplement Standard Reference Materials (SRMs) has been issued by the National Institute of Standards and Technology (NIST) with certified values for ephedrine alkaloids, synephrine, caffeine, and selected toxic trace elements. The materials represent a variety of natural, extracted, and processed sample matrixes that provide different analytical challenges. The constituents have been determined by multiple independent methods with measurements performed by NIST and by three collaborating laboratories. The methods utilized different sample extraction and cleanup steps in addition to different instrumental analytical techniques and approaches to quantification. In addition, food-matrix proximates were determined by National Food Processor Association laboratories for one of the ephedra containing SRMs. The SRMs are primarily intended for method validation and for use as control materials to support the analysis of dietary supplements and related botanical materials. PMID- 15889899 TI - Adsorption equilibria of benzodiazepines on a hybrid polymeric chiral stationary phase. AB - The chromatographic behavior of a series of racemic benzodiazepines was evaluated under linear and nonlinear conditions on a new hybrid polymeric (DACH-ACR) chiral stationary phase (CSP). Differently substituted benzodiazepines were employed as probes to make hypotheses concerning possible molecular interaction mechanisms originating between target compounds and active sites on the CSP. Hydrogen bonds were found to be pivotal for chromatographic retention and chiral selectivity. The competitive effect from a mobile-phase (MP) modifier able to interact with the CSP through H-bonds was investigated. The performance of the polymeric DACH ACR CSP for preparative purposes was also evaluated. The competitive adsorption isotherms of two benzodiazepines, lorazepam and temazepam, were measured at different MP compositions through the so-called inverse method. The adsorption data were fitted with a competitive bi-Langmuir adsorption isotherm. Enantiomeric separations under nonlinear conditions were modeled by using the equilibrium dispersive (ED) model of chromatography. Theoretical overloaded band profiles (obtained by solving the system of partial differential equations described by the ED model) matched, in a significantly accurate way, the profiles experimentally measured. PMID- 15889900 TI - Quantitative 1H NMR with external standards: use in preparation of calibration solutions for algal toxins and other natural products. AB - We examine the use of external standards for quantitative measurement by 1H NMR of solution concentrations of natural products and other low molecular weight, hydrogen-containing compounds and show that precision and accuracy ca. 1% is obtainable with a commercial 11.7 T spectrometer when standards and analytes are contained in separate but identical sealed precision glass NMR tubes. Numerous factors contributing to the intensity of the NMR signals are evaluated. Precise measurements of 360 degrees pulse lengths for each sample provide direct corrections for variations in probe Q-factor that enable samples in different solvents to be compared, provided single-coil excitation and detection is used throughout. Samples need not be prepared in deuterated solvents if the 1H spectra of the solvents are simple enough for peak suppression by presaturation. The approach is particularly suitable for hazardous materials kept in sealed tubes and for the preparation of certified calibration solution reference materials for use with LC-MS and other techniques where deuterated solvents should be avoided. PMID- 15889901 TI - Multiple sampling in single-cell enzyme assays using CE-laser-induced fluorescence to monitor reaction progress. AB - A novel method for assaying enzymes from a single cell or small cell populations is described. The key advantage of this method is the ability to repeatedly sample a single cell enzyme reaction. Whereas multiple sampling has been achieved for larger cell types with a diameter of 1 mm, we report a technique by which single cell enzyme assays of small cells (15 microm in diameter) can be repeatedly carried out. Individual cells were isolated using an in-house-built micromanipulator and placed in nanoliter-scale reaction vessels. The cells were lysed with solution containing substrate, and enzyme activity was assayed by removing 5-nL aliquots with a recently developed nanopipettor. The reaction aliquot was then analyzed using capillary electrophoresis with laser-induced fluorescence detection to quantitate enzyme activity. Sf9 cells were assayed at the single cell level and found to be highly heterogeneous with respect to alpha glucosidase II activity. Since only 5 nL of the single cell reaction was removed, multiple sampling was possible, allowing triplicate analysis of enzyme activity for each individual cell. Multiple sampling also permitted a single cell reaction to be monitored over time. The sensitivity of this method was demonstrated in the analysis of a low-abundance enzyme, alpha1,3-N-acetylgalactosaminyltransferase, from single HT29 cells. Detecting the product of this enzyme reaction required minimizing the dilution of cellular contents. To demonstrate the potential applications of this methodology in small scale biochemical analyses, single Arabidopsis knf embryos lacking the alpha-glucosidase I encoding KNOPF gene were assayed. Mutant embryos demonstrated insignificant conversion of a triglucose substrate, as compared to wild type, confirming the deletion of alpha-glucosidase I. Embryos were simultaneously assayed for a second enzyme, beta-galactosidase, illustrating that the mutants were viable except for their lack of alpha glucosidase I activity. PMID- 15889902 TI - Stochastic theory of size exclusion chromatography: peak shape analysis on single columns. AB - The stochastic theory of size exclusion chromatography (SEC) was applied to analyze the peak shape of chromatograms obtained with a wide range of polystyrene standards on various columns. The columns were packed with stationary phases of different pore sizes. The stochastic-dispersive model of SEC results in a peak shape model that fits well the symmetrical and asymmetrical peaks observed in SEC. From the peak shape parameters obtained after nonlinear parameter estimation, information can be gained regarding the fundamental characteristics of the size exclusion process. When a series of polymer standards are analyzed on one column-in a manner similar to other methods of inverse chromatography-the stationary phase can be characterized. The dependence of the ingress and the egress processes on the relative size of the macromolecule to the pore size was determined. We found that for small molecules the selectivity in SEC arises from the ingress process, while when the size of the macromolecule is comparable to that of the pore-i.e., close to the exclusion limit-the egress process will also strongly affect the selectivity. PMID- 15889903 TI - Uptake of diet resveratrol into the human low-density lipoprotein. Identification and quantification of resveratrol metabolites by liquid chromatography coupled with tandem mass spectrometry. AB - In this paper, a sensitive, precise, and selective analytical method has been developed for the identification and quantification of resveratrol metabolites in human low-density lipoprotein (LDL) after moderate consumption of red wine, using high-performance liquid chromatography electrospray in tandem mass spectrometry (LC-ESI-MS/MS). From different extraction procedures tested, solid-phase extraction was selected to minimize matrix effects reaching the highest sensitivity. Standard calibration curves prepared in human LDL for trans resveratrol were linear over a range of 0.44-438.59 pmol/mL. The accuracy and interassay precision of this LC-MS/MS assay for resveratrol showed a coefficient of variation of <6.0%. The method allows detection and quantification limits for resveratrol in LDL at 0.15 and 0.44 pmol/mL, respectively. Results to date indicate that resveratrol metabolites were incorporated into LDL after a moderate intake of red wine. The metabolites identified in LDL were trans-resveratrol-3-O glucuronide, cis-resveratrol-3-O-glucuronide, and cis-resveratrol-3-O-glucoside, as well as free trans-resveratrol. To our knowledge, it is the first time that a polyphenol from red wine, specifically resveratrol, has been identified in human LDL after moderate intake of red wine. Furthermore, these findings suggest that these compounds may deliver their antioxidant effect to LDL. PMID- 15889904 TI - Continuous real-time analysis of products from the reaction of some monoterpenes with ozone using atmospheric sampling glow discharge ionization coupled to a quadrupole ion trap mass spectrometer. AB - An on-line technique has been demonstrated for the analysis of photochemical oxidation reaction products. The technique is based on the direct introduction of gas and particulate oxidation products into a custom-built atmospheric sampling glow discharge ionization source (ASGDI) coupled to a quadrupole ion trap mass spectrometer (QITMS). Operational parameters of the ASGDI system were investigated to determine their influence on the ion signal for the analysis of oxidation products in real time. These parameters include the discharge current, ion accumulation time, and type of reagent gas. Reference mass spectra from standards were generated for a variety of biogenic compounds and terpene reaction products containing keto, hydroxy, aldehyde, carboxylic acid, or epoxy groups to better understand the fragmentation that occurs in the glow discharge ion source. Results are presented for ozonolysis reactions of four biogenic monoterpenes (alpha-pinene, beta-pinene, D-limonene, Delta(3)-carene) monitored with the ASGDI quadrupole ion trap to demonstrate the ability to obtain real-time measurements. The reaction products identified with ASGDI-QITMS correspond to those products identified with other techniques, including on-line atmospheric pressure chemical ionization techniques. Efficient differentiation of multifunctional products including mono-/di-/hydroxy-/keto-carboxylic acid and keto-/hydroxy-aldehyde was possible by use of the MS/MS capability of the instrument. PMID- 15889905 TI - Discovery, biosynthesis, and structure elucidation of metabolites of a doping agent and a direct analogue, tetrahydrogestrinone and gestrinone, using human hepatocytes. AB - Tetrahydrogestrinone (18a-homo-pregna-4,9,11-trien-17beta-ol-3-one, THG) is an anabolic androgenic steroid sold to athletes as an undetectable performance enhancer. Being an unapproved substance, no legitimate in vivo human excretion studies could be performed to identify urinary markers of this doping agent. In vitro systems were used as an alternative approach to study the human metabolism of THG and the gestrinone analogue, which is a marketed drug. Incubations of both compounds in the presence of human hepatocytes led to formation of oxidative and glucuroconjugated metabolites. Microgram quantities of the major in vitro metabolites were biosynthesized using human hepatocytes, characterized by HPLC/MS/MS, and their structures elucidated by NMR. Due to high structure similarity, both THG and gestrinone had an analogous in vitro metabolic pathway leading to successive addition of a hydroxyl and a beta-glucuronic acid at C-18. This in vitro metabolite of gestrinone was consistent with a previously reported major but unknown human urinary metabolite. The structure of another metabolite of THG was proposed to be a glucuroconjugate of an oxidative product with a hydroxyl group most likely at C-16epsilon. In vitro information reported therein could significantly impact the identification of new urinary markers of THG for doping control purposes. PMID- 15889906 TI - Reagent anions for charge inversion of polypeptide/protein cations in the gas phase. AB - Various reagent anions capable of converting polypeptide cations to anions via ion/ion reactions have been investigated. The major charge inversion reaction channels include multiple proton transfer and adduct formation. Dianions composed of sulfonate groups as the negative charge carriers show essentially exclusive adduct formation in converting protonated peptides and proteins to anions. Dianions composed of carboxylate groups, on the other hand, show far more charge inversion via multiple proton transfer, with the degree of adduct formation dependent upon both the size of the polypeptide and the spacings between carboxylate groups in the dianion. More highly charged carboxylate-containing anions, such as those derived from carboxylate-terminated polyamidoamine half generation dendrimers show charge inversion to give anion charges as high in magnitude as -4, with the degree of adduct formation being inversely related to dendrimer generation. All observations can be interpreted on the basis of charge inversion taking place via a long-lived chemical complex. The lifetime of this complex is related to the strengths and numbers of the interactions of the reactants in the complex. Calculations with model systems are fully consistent with sulfonate groups giving rise to more stable complexes. The kinetic stability of the complex can also be affected by the presence of electrostatic repulsion if it is multiply charged. In general, this situation destabilizes the complex and reduces the likelihood for observation of adducts. The findings highlight the characteristics of multiply charged anions that play roles in determining the nature of charge inversion products associated with protonated peptides and proteins. PMID- 15889907 TI - Amperometric biosensors based on redox polymer-carbon nanotube-enzyme composites. AB - Based on their size and unique electrical properties, carbon nanotubes offer the exciting possibility of developing ultrasensitive, electrochemical biosensors. In this study, we describe the construction of amperometric biosensors based on the incorporation of single-walled carbon nanotubes modified with enzyme into redox polymer hydrogels. The composite films were constructed by first incubating an enzyme in a single-walled carbon nanotube (SWNTs) solution and then cross-linking within a poly[(vinylpyridine)Os(bipyridyl)(2)Cl(2+/3+)] polymer film. Incorporation of SWNTs, modified with glucose oxidase, into the redox polymer films resulted in a 2-10-fold increase in the oxidation and reduction peak currents during cyclic voltammetry, while the glucose electrooxidation current was increased 3-fold to approximately 1 mA/cm(2) for glucose sensors. Similar effects were also observed when SWNTs were modified with horseradish peroxidase prior to incorporation into redox hydrogels. PMID- 15889908 TI - Study of chemistry in droplets with net charge before and after Coulomb explosion: ion-induced nucleation in solution and implications for ion production in an electrospray. AB - Droplets with net charge are essential intermediaries in the production of gaseous ions in the electrospray (ES) ion source. There could be a wealth of knowledge regarding the chemistry that occurs in such droplets as a result of their violation of electroneutrality. Such information could lead to improved understanding of the ion generation process in an ES along with factors that affect it. The experiments performed involved the levitation of individually charged droplets that were, and were not, allowed to undergo Coulomb explosion while they remained levitated in an electrodynamic balance (EDB). Through examination of precipitates formed within the levitated droplets, it was observed that onset of NaCl precipitation was promoted in droplets (glycerol:water 9:1 v/v) that had mass-to-net-charge (m/z) ratio <-4.8 x 10(9) amu/e. This threshold m/z value is exceeded in essentially all droplets generated in an ES because it is above the calculated threshold for Coulomb explosion. This finding suggests that cluster formation in droplets having net charge could occur at solute concentrations lower than would be anticipated on the basis of homogeneous nucleation. The effect of large entities such as precipitates existing in the droplet on the dynamics of droplet Coulomb explosion was also examined. Using droplets whose initial size and magnitude of net charge were equivalent within experimental error but having different concentration of solutes, we showed that the m/z of their main residues following Coulomb explosion were different. Micrometer-size droplets containing 20 nm fluorescent beads that underwent Coulomb explosion resulted in main residues that had higher m/z for higher initial bead concentration in the starting solution (320 nM) when compared to main residues resultant from starting solutions having lower initial bead concentration (21 nM). PMID- 15889909 TI - Evaluating preparative isoelectric focusing of complex peptide mixtures for tandem mass spectrometry-based proteomics: a case study in profiling chromatin enriched subcellular fractions in Saccharomyces cerevisiae. AB - We have evaluated the use of free-flow electrophoresis, an emerging separation method for preparative isoelectric focusing of complex peptide mixtures, as a tool for high-throughput tandem mass spectrometry-based proteomic analysis. In this study, we investigated the ability of free-flow electrophoresis to resolve and fractionate complex peptide mixtures and also the effectiveness of using peptide isoelectric point in conjunction with peptide match probability scoring in sequence database searching. As a model system for this study, we analyzed a chromatin-enriched fraction from the yeast Saccharomyces cerevisiae. This mixture was fractionated using preparative isoelectric focusing by free-flow electrophoresis, followed by online capillary liquid chromatography electrospray tandem mass spectrometry and sequence database searching. Our results demonstrate that (1) FFE effectively resolves and fractionates complex peptide mixtures on the basis of peptide isoelectric point and (2) the introduction of peptide pI is effective in minimizing both false positive and false negative sequence matches in sequence database searching of tandem mass spectrometry data. PMID- 15889910 TI - Imaging of copper, zinc, and other elements in thin section of human brain samples (hippocampus) by laser ablation inductively coupled plasma mass spectrometry. AB - Laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) was used to produce images of element distribution in 20-microm thin sections of human brain tissue. The sample surface was scanned (raster area approximately 80 mm(2)) with a focused laser beam (wavelength 213 nm, diameter of laser crater 50 microm, and laser power density 3 x 10(9) W cm(-2)) in a cooled laser ablation chamber developed for these measurements. The laser ablation system was coupled to a double-focusing sector field ICPMS. Ion intensities of 31P+, 32S+, 56Fe+, 63Cu+, 64Zn+, 232Th+, and 238U+ were measured within the area of interest of the human brain tissue (hippocampus) by LA-ICPMS. The quantitative determination of copper, zinc, uranium, and thorium distribution in thin slices of the human hippocampus was performed using matrix-matched laboratory standards. In addition, a new arrangement in solution-based calibration using a micronebulizer, which was inserted directly into the laser ablation chamber, was applied for validation of synthetic laboratory standard. The mass spectrometric analysis yielded an inhomogeneous distribution (layered structure) for P, S, Cu, and Zn in thin brain sections of the hippocampus. In contrast, Th and U are more homogeneously distributed at a low-concentration level with detection limits in the low nanogram per gram range. The unique analytical capability and the limits of LA ICPMS will be demonstrated for the imaging of element distribution in thin cross sections of brain tissue from the hippocampus. LA-ICPMS provides new information on the spatial element distribution of the layered structure in thin sections of brain tissues from the hippocampus. PMID- 15889911 TI - Preparation of a selective receptor for ephedrine for the development of an optical spot test for the detection of ephedrine in human urine using stabilized in air lipid films with incorporated receptor. AB - The present technique describes the preparation of a selective receptor for ephedrine and a simple sensitive spot optical test for the rapid one-shot detection of ephedrine in human urine using lipid films with an incorporated receptor that are synthesized by a chemical reaction with a methacrylate polymer on a glass fiber filter. The selective receptor was synthesized using a resorcin[4]arene receptor and by transforming all the -OH groups into methoxy groups. The lipid films without this receptor provided fluorescence under a UV lamp. The use of the receptor in these films quenched this fluorescence, and the color became similar to that of the filters without the lipid films. A drop of aqueous solution of ephedrine provided a "switching on" of the fluorescence, which allows the rapid detection of this stimulant in human urine at the levels of 10(-8) M concentrations. The effect of potent interferences (i.e., proteins, lipids, ascorbic aid, glucose, leucine, glycine, tartrate, citrate, bicarbonate, and caffeine) was examined. The results showed no interferences from these compounds in concentration levels usually found in human urine samples. Dopamine was also investigated as a potent interfering agent, and the results have shown that the transformation of the hydroxy to methoxy groups has altered the selectivity of the receptor. This species does not cause interference at concentration levels lower than 10(-6) M. A drop of urine containing ephedrine provided also a "switching on" of the fluorescence, which allows the rapid detection of this stimulant in human urine at the levels of 10(-8) M concentrations. The reproducibility of the method was checked in approximately 100 samples, and all of them were found to provide similar results. Note that the colors of the filters remain stable for periods of more than 2 months. PMID- 15889912 TI - Cross-correlation of optical microcavity biosensor response with immobilized enzyme activity. Insights into biosensor sensitivity. AB - Porous silicon multilayer structures have remarkable optical and morphological properties that can be exploited for biosensing. In particular, a high internal surface area (>100 m(2)/cm(3)) and a linear response profile to changes in the dielectric environment enable fabrication of sensitive devices and a straightforward quantitation of the optical response. These essential operating characteristics are illustrated for p+ mesoporous silicon (pore diameter 15-20 nm) optical microcavities. A series of devices were prepared to permit the immobilization of glutathione-S-transferase ( approximately 50 kDa) within the porous matrix. Enzyme activity was exploited as an indirect means to quantitate the amount of protein immobilized. Activity was positively correlated with the optical sensor response. However, at high enzyme load the activity becomes nonlinear while the microcavity response remains linear. These data were used to determine the transduction limit (minimum amount of protein required to transduce an optical response), which is reported as areal mass sensitivity ranging between 50 and 250 pg/mm(2). This value is considered in context with the dynamic range of the bulk sensitivity, defined as the magnitude of the wavelength shift per refractive index unit, which was measured as a function of microcavity design parameters. This work has uncovered key parameters that can be tuned to improve the detection limit of this sensor modality. Because of the ever increasing number of emerging new biosensor technologies, defining sensor detection limits has become an ambiguous topic and a need exists to standardize measurements and sensitivity units. For chip-based devices, it seems appropriate to report sensitivity in terms of the minimum number of grams of bound target per surface area. PMID- 15889913 TI - Optical metal ion sensor based on diffusion followed by an immobilizing reaction. Quantitative analysis by a mesoporous monolith containing functional groups. AB - A new optical metal ion sensor based on diffusion followed by an immobilizing reaction has been developed. The current sensor is based on a model that unifies two fundamental processes which a metal analyte undergoes when it is exposed to a porous, ligand-grafted monolith: (a) diffusion of metal ions to the binding sites and (b) metal-ligand (ML(n)) complexation. A slow diffusion of the metal ions is followed by their fast immobilizing reaction with the ligands in the monolith to give a complex. Inside the region where the ligands have been saturated, the diffusion of the metal ions reaches a steady state with a constant external metal ion concentration (C(0)). If the complex ML(n) could be observed spectroscopically, the absorbance of the product A(p) follows: A(p) = Kt(1/2), K = 2epsilon(p)(L(0)C(0)D)(1/2). D = diffusion constant of the metal ions inside the porous solid; L(0) = concentration of the ligands grafted in the monolith; and t = time. This equation is straightforward to use, and the K vs C(0)(1/2) plot provides the correlations with the concentrations (C(0)) of the metal ions. This is a rare optical sensor for quantitative metal ion analysis. The use of the model in a mesoporous sol-gel monolith containing grafted amine ligands for quantitative Cu(2+) sensing is demonstrated. This model may also be used in other chemical sensors that depend on diffusion of analytes followed by immobilizing reactions in porous sensors containing grafted/encapsulated functional groups/molecules. PMID- 15889914 TI - Magnetic bead-based chemiluminescent metal immunoassay with a colloidal gold label. AB - A novel, sensitive chemiluminescent (CL) immunoassay has been developed by taking advantage of a magnetic separation/mixing process and the amplification feature of colloidal gold label. First, the sandwich-type complex is formed in this protocol by the primary antibody immobilized on the surface of magnetic beads, the antigen in the sample, and the second antibody labeled with colloidal gold. Second, a large number of Au3+ ions from each gold particle anchored on the surface of magnetic beads are released after oxidative gold metal dissolution and then quantitatively determined by a simple and sensitive Au3+-catalyzed luminol CL reaction. Third, this protocol is evaluated for a noncompetitive immunoassay of a human immunoglobulin G, and a concentration as low as 3.1 x 10(-12) M is determined, which is competitive with colloidal gold-based anodic stripping voltammetry (ASV), colorimetric ELISA, or immunoassays based on fluorescent europium chelate labels. The high performance of this protocol is related to the sensitive CL determination of Au3+ ion (detection limit of 2 x 10(-10) M), which is 25 times higher than that by ASV at a single-use carbon-based screen-printed electrode. From the analytical chemistry point of view, this protocol will be quite promising for numerous applications in immunoassay and DNA hybridization. PMID- 15889915 TI - Ligase detection reaction/hybridization assays using three-dimensional microfluidic networks for the detection of low-abundant DNA point mutations. AB - We have fabricated a flow-through biochip assembly that consisted of two different microchips: (1) a polycarbonate (PC) chip for performing an allele specific ligation detection reaction (LDR) and (2) a poly(methyl methacrylate) (PMMA) chip for the detection of the LDR products using an universal array platform. The operation of the device was demonstrated by detecting low-abundant DNA mutations in gene fragments (K-ras) that carry point mutations with high diagnostic value for colorectal cancers. The PC microchip was used for the LDR in a continuous-flow format, in which two primers (discriminating primer that carried the complement base to the mutation being interrogated and a common primer) that flanked the point mutation and were ligated only when the particular mutation was present in the genomic DNA. The miniaturized reactor architecture allowed enhanced reaction speed due to its high surface-to-volume ratio and efficient thermal management capabilities. A PMMA chip was employed as the microarray device, where zip code sequences (24-mers), which were complementary to sequences present on the target, were microprinted into fluidic channels embossed into the PMMA substrate. Microfluidic addressing of the array reduced the hybridization time significantly through enhanced mass transport to the surface-tethered zip code probes. The two microchips were assembled as a single integrated unit with a novel interconnect concept to produce the flow-through microfluidic biochip. A microgasket, fabricated from an elastomer poly(dimethylsiloxane) with a total volume of the interconnecting assembly of <200 nL, was used as the interconnect between the two chips to produce the three dimensional microfluidic network. We successfully demonstrated the ability to detect one mutant DNA in 100 normal sequences with the biochip assembly. The LDR/hybridization assay using the assembly performed the entire assay at a relatively fast processing speed: 6.5 min for on-chip LDR, 10 min for washing, and 2.6 min for fluorescence scanning (total processing time 19.1 min) and could screen multiple mutations simultaneously. PMID- 15889916 TI - Single DNA molecules as probes of chromatographic surfaces. AB - YOYO-I-labeled lambda-DNA was employed as a nanoprobe for different functionalized surfaces to elucidate adsorption in chromatography. While the negatively charged backbone is not adsorbed, the 12-base unpaired ends of this DNA provide exposed purine and pyrimidine groups for adsorption. Self-assembled monolayers (SAMs) formed on gold substrate provide a wide range of choices of surface with well-defined and well-organized functional groups. Patterns of amino terminated, carboxylic acid-terminated, and hydroxyl-terminated SAMs are generated by lithography. Patterns of metal oxides are generated spontaneously after deposition of metals. By recording the real-time dynamic motion of DNA molecules at the SAMs/aqueous interface, one can study the various parameters governing the retentivity of an analyte during chromatographic separation. Even subtle differences among adsorptive forces can be revealed. PMID- 15889917 TI - Surface-enhanced Raman spectroscopy of self-assembled monolayers: sandwich architecture and nanoparticle shape dependence. AB - Surface enhanced Raman scattering (SERS) spectra of 4-mercaptobenzoic acid (4 MBA) self-assembled monolayers (SAMs) on gold substrates is presented for SAMs onto which gold nanoparticles of various shapes have been electrostatically immobilized. SERS spectra of 4-MBA SAMs are enhanced in the presence of immobilized gold nanocrystals by a factor of 10(7)-10(9) relative to 4-MBA in solution. Large enhancement factors are a likely result of plasmon coupling between the nanoparticles (localized surface plasmon) and the smooth gold substrate (surface plasmon polariton), creating large localized electromagnetic fields at their interface, where 4-MBA molecules reside in this sandwich architecture. Moreover, enhancement factors depend on nanoparticle shape and vary by a factor of 10(2). This SERS geometry offers large surface enhancements for molecules adsorbed onto planar substrates and could be quite useful for determining chemical information for poor Raman scatterers from assays on 2-D substrates. PMID- 15889918 TI - Microelectrode biosensor for real-time measurement of ATP in biological tissue. AB - The purines ATP, ADP, and adenosine are important extracellular signaling agents. Analysis of purinergic signaling has been slowed by lack of direct methods for measurement of purine release in real-time during physiological activity. We have previously reported microelectrode biosensors for adenosine, but similar sensors for ATP have remained elusive. We now describe an ATP biosensor formed by coating a Pt microelectrode with an ultrathin biolayer containing glycerol kinase and glycerol-3-phosphate oxidase. It responds rapidly (10-90% rise time <10 s) and exhibits a linear response to ATP over the physiologically relevant concentrations of 200 nM-50 microM and is very sensitive approximately 250 mA.M( 1).cm(-2). By including phosphocreatine kinase in the biolayer, we can optionally amplify the ATP signal and also make the sensor sensitive to external ADP. We have used our sensors to make the first demonstration that ATP is released from spinal networks in vivo during locomotor activity. PMID- 15889919 TI - Laser-induced dissociation/high-energy collision-induced dissociation fragmentation using MALDI-TOF/TOF-MS instrumentation for the analysis of neutral and acidic oligosaccharides. AB - Producing detailed mass spectrometric fragmentation data of native oligosaccharides for the purpose of basic structure elucidation has become a readily accessible tool since the availability of enhanced technical equipment. In this report, high-energy collision-induced dissociation (heCID) in combination with MALDI-TOF/TOF technology for analysis of native neutral and acidic oligosaccharides is described. By providing complementary data, heCID-MALDI TOF/TOF adds a variety of valuable cross-ring fragmentation information to the information of glycosidic fragmentation obtained preferably by laser-induced dissociation (LID). We examined parameters influencing fragmentation behavior of both-acidic and neutral-compounds. Results show a dependency of the fragmentation pattern for the employed matrix as well as the laser intensity provided for the ionization of the analytes and the complexity of the analytes. Due to instrument specific settings, protonated glycosidic ion series within spectra of sodiated compounds could also be identified. Furthermore, acquired spectra could be readily used to identify compounds by comparison to existing glycan databases such as GlycoSuiteDB and GlycosciencesDB. The results show a better scoring of heCID data sets in comparison to LID-derived data. heCID-MALDI-TOF/TOF analysis in combination with database search algorithms is demonstrated to be suitable for an initial identification/classification of carbohydrates. PMID- 15889920 TI - Microchip immobilized enzyme reactors for hydrolysis of methyl cellulose. AB - Microchip immobilized enzyme reactors (microIMERs) with immobilized endoglucanases were applied for the hydrolysis of methyl cellulose (MC). MCs of various molecular weights were hydrolyzed using two microIMERs containing immobilized celloendoglucanase Cel 5A from Bacillus agaradhaerens (BaCel 5A) connected in series. Hydrolysis by the microIMER could be confirmed from the average molar masses and molar mass distributions measured by size exclusion chromatography (SEC) with online multiangle light scattering and refractive index detection. Methylated cellooligosaccharides with degrees of polymerization (DP) between 1 and 6 formed during hydrolysis were analyzed by direct infusion electrospray ionization ion-trap mass spectrometry (ESI-ITMS). Mass spectra of microIMER- and batch-hydrolyzed samples were compared and no significant differences were found, indicating that microIMER hydrolysis was as efficient as conventional batch hydrolysis. A fast and automated hydrolysis with online MS detection was achieved by connecting the microIMER to high-performance liquid chromatography and ESI-ITMS. This online separation reduced the relative intensities of interfering signals and increased the signal-to-noise ratios in MS. The microIMER hydrolysates were also subjected to SEC interfaced with matrix assisted laser desorption/ionization time-of-flight mass spectrometry. With this technique, oligomers with DP 3-30 could be detected. The hydrolysis by the microIMER was performed within 60 min, i.e. significantly faster compared with batch hydrolysis usually performed for at least 24 h. The microIMER also allowed hydrolysis after 10 days of continuous use. The method presented in this work offers new approaches for the analysis of derivatized cellulose and provides the possibility of convenient online, fast, and more versatile analysis compared with the traditional batch method. PMID- 15889921 TI - Analysis of poly(oxyethylene) and poly(oxypropylene) triblock copolymers by MALDI TOF mass spectrometry. AB - Triblock copolymers of ethylene oxide (EO) and propylene oxide (PO) are widely used in the chemical industry as nonionic surfactants. Triblock copolymers can be arranged in a EO-PO-EO or PO-EO-PO sequence. This arrangement results in an amphiphilic copolymer, in which the block sequence and block length determine the properties of the copolymer. MALDI-TOF MS was used to analyze various triblock copolyethers: EO-PO-EO (Mn =2000 g.mol(-1)), PO-EO-PO (Mn = 2000 g.mol(-1)), and a random copolymer EO/PO (Mn = 2500 g.mol(-1)). Data treatment was assisted by using a homemade software allowing a picture of monomer composition of oligomers from the mass spectra. MALDI-TOF mass spectra of EO/PO copolymers were shown to depend strongly on the number of laser shots, relative proportions of polymer/salt, and the nature of the matrix. An unsaturated byproduct was detected. Its presence was demonstrated by prefractionation of copolymers by SEC before MALDI-TOF analysis, and its content was estimated by 1H NMR. The formation of layers inside the MALDI deposit was evidenced by varying the number of laser shots. Lighter oligomers of the copolymer, unsaturated byproduct, or both would be in the core of the deposit, coated with heavier oligomer. The layer formation depends on the nature of the matrix and the quantity of added salt. DHB matrix with a relative high sodium salt content induces layer formation inside the deposit, whereas dithranol matrix or low salt content does not. Consequently, an optimization of experimental parameters in order to estimate the lighter oligomers or unsaturated byproduct content or to obtain the actual representation of the monomer contribution in the copolymers from the MS data only seems obviously critical. MALDI-TOF mass spectrometry is obviously a powerful technique to analyze copolymers, but a careful survey of the experimental parameters is required. The combination of MALDI-TOF MS with separations techniques and NMR brings precious complementary information. PMID- 15889922 TI - Preparation and evaluation of proline-based chiral columns. AB - Chiral stationary phases made of readily available proline peptides were prepared and evaluated for general chiral separation. With the proper structural elements, these columns demonstrated broad chiral selectivity. Among the 53 analytes tested, a tetraproline column resolved 31. The separations achieved for these analytes are comparable to those achieved on Whelk O2 column, while still inferior to those achieved on Daicel AD-H and OD-H columns. Number of proline units proves important for chiral recognition, because a control column made with a single proline unit is largely ineffective. PMID- 15889923 TI - Microbioassay system for antiallergic drug screening using suspension cells retaining in a poly(dimethylsiloxane) microfluidic device. AB - This article describes an antiallergic drug-screening system by the detection of histamine released from mast cells (suspension cells) on a multilayer microchip. In this study, the elastmeric material, poly(dimethylsiloxane) (PDMS), was employed to fabricate microchannels and microchambers. The microchip consists of two sections: a histamine-releasing one, which has a cell chamber, and a histamine-derivatizing one. Both were laminated to one microchip. Rat peritoneal mast cells were retained in the cell chamber (1.2 microL) with a filtering system using a cellulose nitrate membrane. This filtering system could easily retain suspension cells without cell damage. Mast cells were viable for a sufficient time to conduct the assay on the cell chamber. The cells were stimulated with a chemical release compound 48/80 (C48/80), and then histamine flowed into the lower layer, where it was derivatized to the fluorescent molecules with o phthalaldehyde and its fluorescence was detected on the microchip. This flow system could detect the time course of the histamine release, and this microchip system required only 20 min for the assay. By this integrated system, 51 pmol of histamine released from 500 cells was detected, and the number of cells required for the assay was reduced to 1% compared with conventional bulk systems. By comparing the released histamine levels with and without drugs, their effect could be evaluated. The inhibition ratio of C48/80 induced-histamine release using an antiallergic drug, disodium cromoglicate (DSCG), was related to the concentration of DSCG. This flow system was applicable for antiallergy drug screening by rapid measurement of the inhibition of histamine release from a very small amount of mast cells. PMID- 15889924 TI - Comprehensive assignment of mass spectral signatures from individual Bacillus atrophaeus spores in matrix-free laser desorption/ionization bioaerosol mass spectrometry. AB - We have fully characterized the mass spectral signatures of individual Bacillus atrophaeus spores obtained using matrix-free laser desorption/ionization bioaerosol mass spectrometry (BAMS). Mass spectra of spores grown in unlabeled, 13C-labeled, and 15N-labeled growth media were used to determine the number of carbon and nitrogen atoms associated with each mass peak observed in mass spectra from positive and negative ions. To determine the parent ion structure associated with fragment ion peaks, the fragmentation patterns of several chemical standards were independently determined. Our results confirm prior assignments of dipicolinic acid, amino acids, and calcium complex ions made in the spore mass spectra. The identities of several previously unidentified mass peaks, key to the recognition of Bacillus spores by BAMS, have also been revealed. Specifically, a set of fragment peaks in the negative polarity is shown to be consistent with the fragmentation pattern of purine nucleobase-containing compounds. The identity of m/z = +74, a marker peak that helps discriminate B. atrophaeus from Bacillus thuringiensis spores grown in rich media is [N1C4H12]+. A probable precursor molecule for the [N1C4H12]+ ion observed in spore spectra is trimethylglycine (+N(CH3)3CH2COOH), which produces a m/z = +74 peak when ionized in the presence of dipicolinic acid. A clear assignment of all the mass peaks in the spectra from bacterial spores, as presented in this work, establishes their relationship to the spore chemical composition and facilitates the evaluation of the robustness of "marker" peaks. This is especially relevant for peaks that have been used to discriminate Bacillus spore species, B. thuringiensis and B. atrophaeus, in our previous studies. PMID- 15889925 TI - Gold nanoparticle-catalyzed luminol chemiluminescence and its analytical applications. AB - Gold colloids with nanoparticles of different sizes were found to enhance the chemiluminescence (CL) of the luminol-H2O2 system, and the most intensive CL signals were obtained with 38-nm-diameter gold nanoparticles. UV-visible spectra, X-ray photoelectron spectra, and transmission electron microscopy studies were carried out before and after the CL reaction to investigate the CL enhancement mechanism. The CL enhancement by gold nanoparticles of the luminol-H2O2 system was supposed to originate from the catalysis of gold nanoparticles, which facilitated the radical generation and electron-transfer processes taking place on the surface of the gold nanoparticles. The effects of the reactant concentrations, the size of the gold nanoparticles. and some organic compounds were also investigated. Organic compounds containing OH, NH2, and SH groups were observed to inhibit the CL signal of the luminol-H2O2-gold colloids system, which made it applicable for the determination of such compounds. PMID- 15889926 TI - High-sensitivity ion mobility spectrometry/mass spectrometry using electrodynamic ion funnel interfaces. AB - The utility of ion mobility spectrometry (IMS) for separation of mixtures and structural characterization of ions has been demonstrated extensively, including in biological and nanoscience contexts. A major attraction of IMS is its speed, several orders of magnitude greater than that of condensed-phase separations. Nonetheless, IMS combined with mass spectrometry (MS) has remained a niche technique, substantially because of limited sensitivity resulting from ion losses at the IMS-MS junction. We have developed a new electrospray ionization (ESI)-IMS QTOF MS instrument that incorporates electrodynamic ion funnels at both front ESI IMS and rear IMS-QTOF interfaces. The front funnel is of the novel "hourglass" design that efficiently accumulates ions and pulses them into the IMS drift tube. Even for drift tubes of 2-m length, ion transmission through IMS and on to QTOF is essentially lossless across the range of ion masses relevant to most applications. The rf ion focusing at the IMS terminus does not degrade IMS resolving power, which exceeds 100 (for singly charged ions) and is close to the theoretical limit. The overall sensitivity of the present ESI-IMS-MS system is comparable to that of commercial ESI-MS, which should make IMS-MS suitable for analyses of complex mixtures with ultrahigh sensitivity and exceptional throughput. PMID- 15889927 TI - Capillary-scale frontal affinity chromatography/MALDI tandem mass spectrometry using protein-doped monolithic silica columns. AB - Frontal affinity chromatography (FAC) interfaced with electrospray mass spectrometry (ESI-MS) has been reported as a potential method for screening of compound mixtures against immobilized target proteins. However, the interfacing of bioaffinity columns to ESI-MS requires that the eluent that passes through the protein-loaded column have a relatively low ionic strength to produce a stable spray. Such low ionic strength solvents can cause serious problems with protein stability and may also affect binding constants and lead to high nonspecific binding to the column. Herein, we report on the interfacing of bioaffinity columns to matrix-assisted laser desorption/ionization (MALDI) MS/MS as a new platform for FAC/MS studies. Capillary columns containing a monolithic silica material with entrapped dihydrofolate reductase were used for frontal affinity chromatography of small-molecule mixtures. The output from the column was combined with a second stream containing alpha-cyano-hydoxycinnamic acid in methanol and was deposited using a nebulizer-assisted electrospray method onto a conventional MALDI plate that moved relative to the column via a computer controlled x-y stage, creating a semipermanent record of the FAC run. The use of MALDI MS/MS allowed for buffers with significantly higher ionic strength to be used for FAC studies, which reduced nonspecific binding of ionic compounds and allowed for better retention of protein activity over multiple runs. Following deposition, MALDI analysis required only a fraction of the chromatographic run time, and the deposited track could be rerun multiple times to optimize ionization parameters and allow signal averaging to improve the signal-to-noise ratio. Furthermore, high levels of potential inhibitors could be detected via MALDI with limited ion suppression effects. Both MALDI- and ESI-based analysis showed similar retention of inhibitors present in compound mixtures when using identical ionic strength conditions. The results show that FAC/MALDI-MS should provide advantages over FAC/ESI-MS for high-throughput screening of compound mixtures. PMID- 15889928 TI - DNA electrochemical sensor based on conducting polymer: dependence of the "signal on" detection on the probe sequence localization. AB - We show in this work that it is possible to make selective direct electrochemical hybridization detection of a target strand onto a probe strand immobilized on a conducting polymer modified with a quinone group, which presents cation-exchange properties. This leads to a "signal-on"detection, a unique behavior in comparison to similar systems described in the literature. It is shown that this system is efficient for various probe and target lengths (10-30 bp) and can discriminate a single mismatch. To go further in comprehension of the detection mechanism, a systematic study of the electrochemical response versus the probe sequence localization onto the immobilized strand is performed. For example, a 30-bp target strand is divided into three shorter 10-bp sequences (A-C, respectively), and we investigate the successive hybridization of these 1/3 strands onto the 30 bp probe strand. It is shown that one probe strand can be used to address several shorter targets. PMID- 15889929 TI - Determination of kinetic and thermodynamic parameters of surface confined species through ac voltammetry and a nonstationary signal processing technique: the Hilbert transform. AB - Data analysis of voltammetric responses has usually been done through application of the fast Fourier transform although it is widely accepted that electrochemical signals are intrinsically nonlinear and nonstationary. In this work, we present a time-series analysis based on the Hilbert transform (HT), a nonstationary signal processing technique, as an alternative tool that can overcome many of the difficulties associated with Fourier techniques. We use the HT to study the behavior of thin-film processes when the excitation perturbation is ac voltammetry. From the analysis of simulated data, we propose simple relations that enable species-specific kinetic and thermodynamic parameters to be estimated, without prior utilization of baseline subtraction even when double layer capacitance significantly influences the current response. We also propose a method to determine whether the characteristics of the applied voltage perturbation are adequate for the accurate estimation of these parameters. The methodology developed here will be applied to previously published experimental time series data (Guo, S. X.; Zhang, J.; Elton, D. M.; Bond, A. M. Anal. Chem. 2004, 76, 166-177.) obtained with ac voltammetry to show how a number of physical parameters can be directly extracted from the processed data. PMID- 15889930 TI - Method for analysis of polar volatile trace components in aqueous samples by gas chromatography. AB - A new method has been developed for direct analysis of volatile polar trace compounds in aqueous samples by gas chromatography. Water samples are injected onto a short packed precolumn containing anhydrous lithium chloride. A capillary column is coupled in series with the prefractionation column for final separation of the analytes. The enrichment principle of the salt precolumn is reverse to the principles employed in conventional methods such as SPE or SPME in which a sorbent or adsorbent is utilized to trap or concentrate the analytes. Such methods are not efficient for highly polar compounds. In the LiCl precolumn concept, the water matrix is strongly retained on the hygroscopic salt, whereas polar as well as nonpolar volatile organic compounds show very low retention and are eluted ahead of the water. After transfer of the analytes to the capillary column, the retained bulk water is removed by backflushing the precolumn at elevated temperature. For direct injections of 120 microL of aqueous samples, the combined time for injection and preseparation is only 3.5 min. With this procedure, direct repetitive automated analyses of highly volatile polar compounds such as methanol or tetrahydrofuran can be performed, and a limit of quantification in the low parts-per-billion region utilizing a flame ionization detector is demonstrated. PMID- 15889931 TI - N-terminal adducts of bovine hemoglobin with glutaraldehyde in a hemoglobin-based oxygen carrier. AB - Hemoglobin-based oxygen carriers (HBOCs) are being developed for the medical field, but because they could increase an athlete's performance, they are misapplied for doping purposes. We previously presented a screening method to detect Oxyglobin (Biopure Corp.) and PolyHeme (Northfield Laboratories Inc.) in serum samples using total acid hydrolysis followed by electrospray mass spectrometry analyses. An alternative mass spectrometric method involving enzymatic hydrolysis is here presented. Digestion of Oxyglobin by endoproteinase Glu-C and LC/MS analyses of the mixture allowed the detection of unique peptidic fragments in comparison with a bovine hemoglobin digest. Tandem mass spectrometry experiments of these peptide ions were performed, and two specific species were actually identified as the N-terminal enzymatic fragment of the beta chain carrying two different modifications. Sequential MS3 experiments using an ion trap mass spectrometer permitted us to locate the chemical modification by the glutaraldehyde on the NH2-terminal group and to propose a structure for the modified peptides. In another set of experiments, screening of these two diagnostic ions into Oxyglobin-spiked serums using precursor ion scan mode in a triple quadrupole instrument allowed the detection of this HBOC with a detection limit of 2 g L(-1). PMID- 15889932 TI - Distribution of gallium nanocrystals in Ga/MCM-41 mesocomposites by continuous flow hyperpolarized 129Xe NMR spectroscopy. AB - The distribution of gallium nanocrystals in mesoporous MCM-41 host were analyzed by continuous-flow hyperpolarized 129Xe NMR spectroscopy. In contrast to unclear TEM images for the high metal contents, laser-polarized 129Xe probe can detect the whole distribution of gallium in the MCM-41 host. It is found that gallium nanocrystals are included in the mesochannels of MCM-41; a part of them also remains in the interparticle voids. The distribution of gallium metal in MCM-41 is heterogeneous. Not all the mesochannels host metallic gallium even at a high gallium loading of 65.1 wt %. Variable temperature measurements can provide information on the xenon adsorption parameters. This approach opens a sensitive way to probe the distribution of high content species in porous host materials. PMID- 15889933 TI - Analysis of derivatized biogenic aldehydes by LC tandem mass spectrometry. AB - Lipid peroxidation has been linked to the etiology of several diseases, including Alzheimer's disease (AD). End products of this phenomenon include low molecular weight, water-soluble aldehydes, compounds that covalently modify proteins and nucleic acids, thereby altering function. Aliphatic aldehydes (C3-C10) are generated during lipid peroxidation, along with alpha,beta-unsaturated aldehydes, including acrolein and 4-hydroxynonenal (HNE). The Hantzsch reaction was used to produce heterocyclic aldehyde derivatives that can be conveniently analyzed with mass spectrometry. Liquid chromatographic analyses revealed increasing retention times from derivatized methanal to octanal. HNE derivatives were observed to elute between heptanal and octanal derivatives, while the acrolein derivatives had a retention time similar to the propanal derivative. Smaller aliphatic aldehyde derivatives fragmented in a similar manner to produce a base peak of m/z 273, while the larger derivatives yielded m/z 274 as the base peak. Acrolein and HNE derivatives fragmented in a slightly different manner compared to their aliphatic counterparts. Calibration plots of aliphatic and unsaturated aldehydes were linear (r2 >/= 0.99) in the concentration range explored (approximately 5 1500 pg on column). The LC-MS/MS methodology developed here will be used in subsequent studies to determine aldehyde concentrations for comparing age-matched controls to AD tissues from human subjects. PMID- 15889934 TI - Measurement of cosmogenic 35S activity in rainwater and lake water. AB - Although cosmogenic 35S (t(1/2) = 87 d) has been found to be a unique and excellent radioactive tracer for stable S in atmospheric and aqueous environments, its application has been very limited because an analytical method for its detection has not been well-documented. Here, we report a rapid and robust method for analyzing extremely low levels of 35S in rainwater and lake water samples. About 20-L water samples were preconcentrated using an anion exchange column. The purified 35S was precipitated as BaSO4, and the precipitates were collected using a GF/B filter. The 35S in precipitates on the filter was directly counted using a super-low-background liquid scintillation counter with cocktail. We successfully measured 35S in precipitation and lake water samples using this method, which promises future diverse applications of the 35S tracer to S cycling in the environment and to age determination of lake water and shallow groundwater. PMID- 15889935 TI - Measuring the mass of an electron by LC/TOF-MS: a study of "twin ions". AB - While investigating the in-source CID fragmentation of nonsteroidal antiinflammatory drugs (NSAIDs), it was noticed that the same fragment ion (nominal mass) formed in either positive or negative ion electrospray for a suite of NSAIDs. For example, ibuprofen with a molecular mass of 206, fragments to the m/z 161 ion in negative ion from its deprotonated molecule (m/z 205, [M - H]-) and fragments to the m/z 161 ion in positive ion from its protonated molecule (m/z 207, [M + H]+). This fragment ion was euphemistically called a "twin ion"because of the same nominal mass despite opposite charge. The CID fragmentation of the twin ions was confirmed also by LC/MS/MS ion trap. Accurate mass measurements in negative ion show that the loss was due to CO2 (measured loss of 43.9897 atomic mass units (u) versus calculated loss of 43.9898 u for N = 10) and in positive ion the loss is due to HCOOH (measured loss of 46.0048 u versus calculated loss of 46.0055 u, N = 10). It was realized that, in fact, the ions were not "identical mass twins of opposite charge" but separated in accurate mass by two electrons. The accurate mass measurement by liquid chromatography/time-of-flight-mass spectrometry (LC/TOF-MS) can distinguish between the two fragment ions of ibuprofen (161.13362 +/- 0.00019 and 161.13243 +/- 0.00014 for N = 20). This experiment was repeated for two other NSAIDs, and the mass of an electron was measured as the difference between the twin ions, which was 0.00062 u +/- 14.8% relative standard deviation (N = 20 analyses). Thus, the use of continuous calibration makes it possible to measure the mass of an electron within one significant figure using the NSAID solution. This result shows the importance of including electron mass in accurate mass measurements and the value of a benchmark test for LC/TOF-MS mass accuracy. PMID- 15889936 TI - Human low-density lipoprotein-coated capillaries in electrochromatography. AB - Low-density lipoprotein (LDL) particles were immobilized on the inner wall of a fused-silica capillary and used in a study of the interactions between LDL and neutral drugs in electrochromatography. The effect of coating parameters (pH, ionic strength of the coating solution, duration of the coating procedure) on the properties and stability of the coating was examined. The stability of the coating was highest when the pH of the coating solution was under the pI value of the LDL particles. Interactions of unmodified LDL coatings with drugs were compared with those of acetylated LDL coatings. Acetylation of LDL neutralizes the positive charge on the lysine residues of the protein component of LDL particles, and acetylated LDL was used as a reference to examine the effect of the positively charged amino acids in the unmodified coating. Under similar coating conditions, acetylated LDL coating yielded stronger EOF evidently due to the decreased number of positive charges on LDL particles. The interactions of the unmodified and acetylated LDL coatings with steroids aldosterone, testosterone, and progesterone were comparable, which indicates that the density of immobilized LDL particles is not appreciably altered by acetylation. As expected, the strength of the interactions between steroids and the LDL coating increased with hydrophobicity of the drug. PMID- 15889937 TI - Mass spectral analysis of chloropicrin under negative ion chemical ionization conditions. AB - A chemical ionization (CI) method is developed for the first time to obtain molecular weight information for chloropicrin (CP), which is used as a chemical warfare agent and as an insecticide. The study includes a detailed investigation on the behavior of CP under electron impact (EI) and CI. Reagent gases of different nature, i.e., methane, isobutane, ammonia, hydrogen, and carbon dioxide, were used for CI analysis. Negative ion mode is found more sensitive than positive ion mode for the EI/CI mass spectrometric analysis of CP, but none of the methods provided molecular weight information, except negative ion CI using ammonia as the reagent gas (NICI (NH3)). The NICI (NH3) showed formation of the quasi-molecular ion, [M + H]-, in addition to other adduct ions. The [M + H]- abundance critically depends on the source temperature, reagent gas pressure, and concentration of the analyte, and it can be 13% under optimized conditions by which CP can be confirmed unambiguously. This method meets the criteria used in official proficiency tests conducted by OPCW for confirming the molecular weight of the unknowns. PMID- 15889938 TI - Parallel ion parking: improving conversion of parents to first-generation products in electron transfer dissociation. AB - Electron-transfer dissociation (ETD) in a tandem mass spectrometer is an analytically useful ion/ion reaction technique for deriving polypeptide sequence information, but its utility can be limited by sequential reactions of the products. Sequential reactions lead to neutralization of some products, as well as to signals from products derived from multiple cleavages that can be difficult to interpret. A method of inhibiting sequential ETD fragmentation in a quadrupole ion trap is demonstrated here for the reaction of a triply protonated peptide with nitrobenzene anions. A tailored waveform (in this case, a filtered noise field) is applied during the ion/ion reaction time to accelerate simultaneously first-generation product ions and thereby inhibit their further reaction. This results in a approximately 50% gain in the relative yield of first-generation products and allows for the conversion of more than 90% of the original parent ions into first-generation products. Gains are expected to be even larger when higher charge-state cations are used, as the rates of sequential reaction become closer to the initial reaction rate. PMID- 15889939 TI - Funding, conflicts, and paradigms. PMID- 15889940 TI - A 4-fold risk of metabolic syndrome in patients with schizophrenia: the Northern Finland 1966 Birth Cohort study. AB - OBJECTIVE: Schizophrenia is associated with a shortened life expectancy and increased somatic comorbidity with, e.g., cardiovascular disorders. One major risk factor for these disorders is the metabolic syndrome, which has been reported to have a higher frequency in schizophrenic patients. Our objective was to study the prevalence of metabolic syndrome in a population-based birth cohort. METHOD: The study sample consisted of 5613 members of the Northern Finland 1966 Birth Cohort who participated in the field study from 1997 to 1998. Subjects were divided into 4 diagnostic categories (DSM-III-R): (1) schizophrenia (N = 31), (2) other functional psychoses (N = 22), (3) nonpsychotic disorders (N = 105), and (4) no psychiatric hospital treatment (N = 5455, comparison group). Subjects were assessed for the presence of metabolic syndrome according to the criteria of the National Cholesterol Education Program. RESULTS: The prevalence of metabolic syndrome was higher in subjects with schizophrenia compared with the comparison group (19% vs. 6%, p = .010). The prevalence of metabolic syndrome in subjects with other psychoses was 5%. After controlling for sex, the results of logistic regression analysis showed that the risk of metabolic syndrome in schizophrenia was 3.7 (95% CI = 1.5 to 9.0). CONCLUSIONS: The high prevalence of metabolic syndrome in schizophrenia even at such a relatively young age underscores the need to select antipsychotic medications with no or little capability to induce metabolic side effects. Also, developing comprehensive efforts directed at controlling weight and diet and improving physical activity are needed. PMID- 15889941 TI - Comorbidity of DSM-IV pathological gambling and other psychiatric disorders: results from the National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVE: To present nationally representative data on lifetime prevalence and comorbidity of pathological gambling with other psychiatric disorders and to evaluate sex differences in the strength of the comorbid associations. METHOD: Data were derived from a large national sample of the United States. Some 43,093 household and group quarters residents age 18 years and older participated in the 2001-2002 survey. Prevalence and associations of lifetime pathological gambling and other lifetime psychiatric disorders are presented. The diagnostic interview was the National Institute on Alcohol Abuse and Alcoholism Alcohol Use Disorder and Associated Disabilities Interview Schedule-DSM-IV Version. Fifteen symptom items operationalized the 10 pathological gambling criteria. RESULTS: The lifetime prevalence rate of pathological gambling was 0.42%. Almost three quarters (73.2%) of pathological gamblers had an alcohol use disorder, 38.1% had a drug use disorder, 60.4% had nicotine dependence, 49.6% had a mood disorder, 41.3% had an anxiety disorder, and 60.8% had a personality disorder. A large majority of the associations between pathological gambling and substance use, mood, anxiety, and personality disorders were overwhelmingly positive and significant (p < .05), even after controlling for sociodemographic and socioeconomic characteristics. Male sex, black race, divorced/separated/widowed marital status, middle age, and living in the West and Midwest were associated with increased risk for pathological gambling. Further, associations between alcohol dependence, any drug use disorder, drug abuse, nicotine dependence, major depressive episode, and generalized anxiety disorder and pathological gambling were stronger among women than men (p > .05). CONCLUSION: Pathological gambling is highly comorbid with substance use, mood, anxiety, and personality disorders, suggesting that treatment for one condition should involve assessment and possible concomitant treatment for comorbid conditions. PMID- 15889942 TI - An open-label study of amisulpride in the treatment of mania. AB - BACKGROUND: Amisulpride is a selective D(2)-D(3) antagonist that has been reported to be effective in the treatment of schizophrenia and major depressive disorder. However, no prospective study to date has assessed the effectiveness and tolerability of this compound in mania. METHOD: Twenty DSM-IV-defined acutely ill manic bipolar patients with a Young Mania Rating Scale (YMRS) score of 20 or more entered this open, prospective, 6-week study. Assessments included the YMRS, the Hamilton Rating Scale for Depression (HAM-D), the Clinical Global Impressions Scale for Bipolar Disorder, Modified (CGI-BP-M), and the systematic report of adverse events. Amisulpride was added to other medications, but other antipsychotics were not allowed. RESULTS: Fourteen patients (70%) completed the study. Using last-observation-carried-forward (LOCF) analyses, amisulpride produced significant improvements on the YMRS (p = .0001), the HAM-D (p < .0141), and the overall (p = .0003), mania (p = .0001), and depression (p = .0268) subscales of the CGI-BP-M. The most common side effect was sedation (N = 5, 25%), but there were also some extrapyramidal symptoms, galactorrhea, insomnia, and agitation. The mean amisulpride dose was 680 mg/day (LOCF) and 786 mg/day in completers. CONCLUSIONS: This first prospective study on amisulpride in the treatment of mania suggests that, despite the limitations of the open, observational design and small sample size, amisulpride may be effective and reasonably safe in the treatment of bipolar mania. D(2) and D(3) antagonism may be involved in the mechanisms of the therapeutic response to antipsychotics in mania. PMID- 15889943 TI - Risk factors for completed suicide in schizophrenia. AB - OBJECTIVE: Schizophrenic patients in Taiwan have lower comorbidity of substance use disorders than do those in Western countries, and most of them live with their families. This study investigated the risk factors for completed suicide in this population with inherently lower rates of the confounding variables of substance abuse and social isolation. METHOD: 4237 acute inpatients with DSM-III, DSM-III-R, or DSM-IV schizophrenia admitted from January 1, 1985, to December 31, 2000, were followed through 2001 by record linkage to the Death Certification System. Seventy-eight subjects who died from suicide during this period were matched with living controls randomly for age (+/- 5 years), sex, and the same year of index admission. Demographic and clinical variables were collected from medical records and formally confirmed at every admission and outpatient follow up. RESULTS: Among 78 case-control pairs, the lifetime prevalence of substance use disorders was 7.1%, and 93.6% of the subjects lived with their families. Approximately half of the completed suicides occurred within 4 years after the first admission. Conditional logistic regression analysis revealed a strong association with the following 3 variables: depressive syndrome in residual phase (adjusted odds ratio [OR] = 23.07, p < .005), higher suicide intensity (adjusted OR = 2.78, p < .05), and later age at onset (increase per year, adjusted OR = 1.07, p < .05). Fasting cholesterol level and clozapine use had no association with completed suicide. CONCLUSIONS: The peak period for completed suicide was early years after the first admission. The target population for additional measures to prevent suicide should include patients with depressive syndrome in residual phase, higher suicide intensity, and later onset of illness. PMID- 15889944 TI - Controlled double-blind trial of phenytoin vs. fluoxetine in major depressive disorder. AB - BACKGROUND: Phenytoin was the first non-sedative anticonvulsant introduced and is still the anticonvulsant most widely used worldwide in neurology. Given the efficacy of the anticonvulsant lamotrigine in the depressed phase of bipolar disorder, a critical theoretical question is whether other anticonvulsants used in treating bipolar disorder might be similarly effective. We therefore undertook a controlled trial of phenytoin versus fluoxetine in major depressive disorder. METHOD: Data were collected from July 2001 to July 2003. Thirty-three subjects entered the study. All patients met DSM-IV criteria for major depressive disorder and scored a minimum of 18 on the 24-item Hamilton Rating Scale for Depression (HAM-D) at baseline. After a 3-day washout of any previous medications, patients were randomly assigned to fluoxetine or phenytoin in identical capsules. Each capsule contained phenytoin 100 mg or fluoxetine 7 mg plus cornstarch. Patients started with 1 tablet daily and increased every other day until they were taking 1 tablet 3 times daily with meals. Blood phenytoin levels were taken after 1 week, 3 weeks, and 6 weeks, and dosage was adjusted to achieve blood levels of 10 to 20 microg/mL, to a maximum dose of 4 capsules per day or a minimum dose of 2 capsules per day. Fluoxetine patients were assigned dummy blood phenytoin levels by the control psychiatrist such that the treating physician would raise the number of capsules to at least 3 per day (20 mg of fluoxetine). RESULTS: Thirty three patients entered the study, and 28 (N = 14 in each treatment group) completed at least 3 weeks and were included in the data analysis. Patients who dropped out after week 3 (3 patients) were included in the study as last value carried forward. There was no difference between treatment groups in overall rate of response or speed of response. CONCLUSION: The absence of a placebo arm in our study allows for the possibility that neither treatment was more effective than placebo. However, the exclusion of past fluoxetine nonresponders and the minimum HAM-D score at baseline of 18 make this possibility unlikely. PMID- 15889945 TI - Pain predicts longer time to remission during treatment of recurrent depression. AB - INTRODUCTION: Pain and depression are mutually exacerbating. We know that both of these syndromes predict the future occurrence of the other. It has not been shown, however, whether the presence of pain slows the effect of treatment for depression. We hypothesized that greater pain and somatic scores prior to treatment with imipramine and interpersonal psychotherapy would predict a slowed time to remission from depression. METHOD: We performed secondary data analyses of an archived study. Subjects (N = 230) were between 21 and 65 years of age and were enrolled in a study of maintenance treatment for recurrent unipolar depression. Patients had to meet Research Diagnostic Criteria (RDC) for a major depressive episode and historical requirements for at least 3 prior episodes and clear remissions (according to RDC). Patients were also required to have a minimum Hamilton Rating Scale for Depression score of 15 and a minimum score of 7 on the Raskin Severity of Depression Scale. This report describes the acute treatment phase, during which all subjects received combination therapy consisting of imipramine hydrochloride (150 to 300 mg) and interpersonal psychotherapy. Pain and somatization were measured with the Hopkins Symptom Checklist. RESULTS: Higher levels of both pain and somatization predicted a longer time to remission. After controlling for baseline severity of depression, only pain was still significant in predicting a longer time to remission. Headache and muscle soreness were the 2 variables from the pain index whose presence independently predicted a slower remission. Both pain and somatization improved during acute treatment. Subjects with more pain and somatization, after controlling for severity of depression, reported more suicidality. Women reported more pain than men. CONCLUSIONS: Pain, but not somatization, predicted a longer time to remission and may be a marker of a more difficult-to-treat depression. Adults with recurrent depression should be screened for the presence of pain prior to treatment, as the presence of these symptoms may require more aggressive treatment or may be a marker for suicidality or the use of dual-mechanism antidepressants. PMID- 15889946 TI - An 8-week open-label trial of a 6-day course of mifepristone for the treatment of psychotic depression. AB - OBJECTIVE: Several investigations suggest that mifepristone leads to the rapid amelioration of psychotic depression. However, these studies were of short duration (1 week or less) and included subjects who were taking other psychotropic medications. The goals of this study were to extend these findings by conducting an 8-week trial of mifepristone for subjects with psychotic depression who were taking no concomitant psychiatric medications. METHOD: Twenty subjects with a DSM-IV major depressive episode with psychotic features (for convenience we use the term psychotic depression) taking no psychotropic medications were given a 6-day course of mifepristone and followed as inpatients for a total of 8 weeks. Nonblinded ratings using the Hamilton Rating Scale for Depression (HAM-D) and Clinical Global Impressions scale (CGI) were performed at baseline and at the end of weeks 1, 4, and 8. The Brief Psychiatric Rating Scale (BPRS) was also administered at baseline and after weeks 4 and 8. Subjects were recruited between February 2003 and December 2003. RESULTS: Significant improvements in HAM-D and CGI scores were shown after 1 week and between weeks 1 and 4 but not between weeks 4 and 8. BPRS scores improved significantly after week 4, while the improvement in BPRS scores between weeks 4 and 8 was of borderline significance. CONCLUSION: Mifepristone appears to be a useful intervention for psychotic depression, leading to significant improvements even after a 1-week course of administration. Issues related to its optimal dosing and to prediction of response are discussed, as are the implications of lack of a placebo group and the use of nonblinded ratings in the present study. PMID- 15889947 TI - Why isn't bupropion the most frequently prescribed antidepressant? AB - OBJECTIVE: Reviews of antidepressant medication efficacy suggest that all antidepressants are equally effective. Bupropion is less likely than other antidepressants to cause weight gain and sexual dysfunction, the 2 side effects that are of greatest concern to patients and that have the greatest impact on long-term compliance. If bupropion is as effective as other antidepressants, and it does not cause the side effects that are the most frequent causes of long-term noncompliance, then why isn't it the most frequently prescribed antidepressant medication? To understand psychiatrists' decision making at the time an antidepressant is chosen, we conducted the Rhode Island Factors Associated With Antidepressant Choice Survey (FAACS). METHOD: For 1137 DSM-IV-diagnosed depressed patients initiated on an antidepressant, the treating psychiatrist completed a 43 item questionnaire listing factors that might have influenced the choice of medication. The questionnaire was filled out immediately after the antidepressant was prescribed to treat a depressive disorder. This study was conducted from August 2001 to February 2002. RESULTS: Because the reasons for choosing a medication to augment an existing regimen might be different from those used in monotherapy, augmentation trials were excluded from the analysis, leaving a sample of 965 patients. Bupropion was rarely prescribed when the presence of comorbid anxiety disorders or symptoms reflecting central nervous system activation influenced antidepressant selection. When the desire to avoid side effects, especially sexual dysfunction and weight gain, were the basis of selection, then bupropion was significantly more often prescribed than other antidepressants (p < .001). CONCLUSIONS: Although there is little evidence that patient factors predict differential medication response, psychiatrists are strongly inclined to base antidepressant selection on clinical profiles and avoid prescribing bupropion for depressed patients with high anxiety. Possible reasons for the discrepancy between psychiatrists' prescribing habits and the results of empirical study are discussed. PMID- 15889948 TI - Analyses of treatment-emergent mania with olanzapine/fluoxetine combination in the treatment of bipolar depression. AB - BACKGROUND: Treatment-emergent mania is a potential risk when patients with bipolar disorder are treated with antidepressant agents. These subanalyses compare treatment-emergent mania rates in bipolar I depressed patients treated with olanzapine, placebo, or olanzapine/fluoxetine combination. METHOD: In this 8 week, double-blind investigation, patients with bipolar I depression (DSM-IV criteria) (N = 833, baseline Montgomery-Asberg Depression Rating Scale total score > or = 20) were randomly assigned to olanzapine (5-20 mg/day, N = 370), placebo (N = 377), or olanzapine/fluoxetine combination (6/25, 6/50, or 12/50 mg/day; N = 86). Treatment-emergent mania was evaluated with the Young Mania Rating Scale (YMRS), the Clinical Global Impressions-Bipolar Edition (CGI-BP) Severity of Mania scale, and adverse events records. RESULTS: Overall rates of study discontinuation due to mania were low and not significantly different among the therapy groups (p = .358). Incidence of treatment-emergent mania (defined as a YMRS score < 15 at baseline and > or = 15 at any subsequent visit) did not differ significantly among therapy groups (olanzapine 5.7%, placebo 6.7%, olanzapine/fluoxetine combination 6.4%; p = .861). Subjects receiving olanzapine or olanzapine/fluoxetine combination had greater mean decreases in YMRS scores than those receiving placebo (p < .001 for both). Subjects receiving olanzapine or olanzapine/fluoxetine combination also had greater mean decreases in CGI-BP scores than those receiving placebo (p = .040 and p = .003, respectively). CONCLUSION: These results suggest that olanzapine/fluoxetine combination does not present a greater risk of treatment-emergent mania compared to olanzapine or placebo over 8 weeks of acute treatment for bipolar I depression. Due to the cyclical nature of bipolar disorder, patients taking olanzapine/fluoxetine combination for bipolar depression should still be monitored for signs or symptoms of emerging mania. PMID- 15889949 TI - Open-label adjunctive zonisamide in the treatment of bipolar disorders: a prospective trial. AB - BACKGROUND: The response of 62 outpatients with DSM-IV bipolar disorders to open label adjunctive zonisamide was evaluated in a prospective 8-week acute trial, followed by a 48-week continuation trial, conducted from June 2001 through May 2002. METHOD: During the acute trial, response to zonisamide was assessed weekly for the first 4 weeks and every 2 weeks for the second 4 weeks with the Clinical Global Impressions scale modified for bipolar illness (CGI-BP), the Young Mania Rating Scale (YMRS), and the Inventory for Depressive Symptomatology (IDS). During the continuation trial, patients were assessed with these scales every 4 weeks. Patients' weights and side effects were also evaluated. Outcome measures were analyzed with repeated-measures analyses of variance. RESULTS: Patients with manic symptoms at study entry (N = 34) displayed significant reductions in CGI-BP Mania Severity and YMRS scores in the acute and continuation (N = 19) trials (p values < .0001 and < .001, respectively). Patients with depressive symptoms at study entry (N = 22) showed significant decreases in CGI-BP-Depression Severity and IDS scores in the acute trial (p values < .001 and < .05, respectively), but only 9 patients entered the continuation trial. Among these 9 patients, maintenance of anti-depressant response was mostly maintained. Initially euthymic patients (N = 6) showed no change in any rating scale scores acutely, but 2 of 4 patients who entered the continuation trial developed depressive symptoms. The 62 patients as a group showed significant weight loss in both trials (p values < .001). However, 20 patients (32%) discontinued zonisamide for worsening mood symptoms. CONCLUSION: Adjunctive zonisamide was associated with beneficial effects on mood and body weight in some patients with bipolar disorders, but was also associated with a high discontinuation rate due to worsening mood symptoms. Double-blind, placebo-controlled studies are necessary to determine zonisamide's thymoleptic properties, if any, in bipolar disorders. PMID- 15889950 TI - Prospective observational study of treatments for unexplained chronic fatigue. AB - BACKGROUND: Unexplained chronic fatigue is a frequent complaint in primary care. A prospective observational study design was used to evaluate whether certain commonly used therapies for unexplained chronic fatigue may be effective. METHOD: Subjects with unexplained chronic fatigue of unknown etiology for at least 6 months were recruited from the Wisconsin Chronic Fatigue Syndrome Association, primary care clinics, and community chronic fatigue syndrome presentations. The primary outcome measure was change in a 5-question fatigue score from 6 months to 2 years. Self-reported interventions tested included prescribed medications, non prescribed supplements and herbs, lifestyle changes, alternative therapies, and psychological support. Linear regression analysis was used to test the association of each therapy with the outcome measure after adjusting for statistically significant prognostic factors. RESULTS: 155 subjects provided information on fatigue and treatments at baseline and follow-up. Of these subjects, 87% were female and 79% were middle-aged. The median duration of fatigue was 6.7 years. The percentage of users who found a treatment helpful was greatest for coenzyme Q10 (69% of 13 subjects), dehydroepiandrosterone (DHEA) (65% of 17 subjects), and ginseng (56% of 18 subjects). Treatments at 6 months that predicted subsequent fatigue improvement were vitamins (p = .08), vigorous exercise (p = .09), and yoga (p = .002). Magnesium (p = .002) and support groups (p = .06) were strongly associated with fatigue worsening from 6 months to 2 years. Yoga appeared to be most effective for subjects who did not have unclear thinking associated with the fatigue. CONCLUSION: Certain alternative therapies for unexplained chronic fatigue, especially yoga, deserve testing in randomized controlled trials. PMID- 15889951 TI - Neuroleptic sensitivity in Parkinson's disease and parkinsonian dementias. AB - BACKGROUND: Severe sensitivity to neuroleptic agents is a major clinical problem in dementia with Lewy bodies (DLB), but has not been determined in Parkinson's disease (PD) and PD with dementia (PDD). METHOD: Severe neuroleptic sensitivity reactions (NSRs) were evaluated according to an operationalized definition blind to clinical and neuropathologic diagnoses in prospectively studied patients exposed to neuroleptics from 2 centers. The study was conducted from June 1995 to May 2003. RESULTS: Ninety-four patients were included (15 with DLB, 36 with PDD, 26 with PD, 17 with Alzheimer's disease, all diagnosed with various operational criteria). Severe NSR only occurred in patients with Lewy body disease: DLB (8 [53%]), PDD (14 [39%]), and PD (7 [27%]), but did not occur in Alzheimer's disease (p = .006). Severe NSR was not associated with other clinical or demographic features. In DLB, severe NSR was not associated with neuropathologic indices (Consortium to Establish a Registry for Alzheimer's Disease staging, Braak staging, or cortical distribution of Lewy bodies). CONCLUSIONS: An operationalized evaluation of severe NSR blind to diagnosis confirmed the high prevalence in DLB and identified high frequencies in Parkinson's disease and PDD with important implications for clinical practice. PMID- 15889952 TI - Long-term maintenance therapy with quetiapine versus haloperidol decanoate in patients with schizophrenia or schizoaffective disorder. AB - OBJECTIVE: To compare the long-term efficacy and tolerability of oral quetiapine with those of intramuscular haloperidol. METHOD: Patients with DSM-IV-diagnosed schizophrenia or schizoaffective disorder requiring long-term antipsychotic treatment were randomly assigned to open-label oral quetiapine or intramuscular haloperidol decanoate for 48 weeks. Clinicians were instructed to target dosing at 500 mg/day of quetiapine or 200 mg of haloperidol decanoate every 4 weeks. The Positive and Negative Syndrome Scale was used to assess efficacy; the Simpson Angus Scale and the Barnes Akathisia Scale were used to assess safety and tolerability. For statistical analyses, a general linear mixed-model repeated measures analysis of covariance was used, with change scores for dependent variables computed with the baseline score as covariate. Data were collected from 1998 to 2001. RESULTS: Thirty-five patients were enrolled, but 6 did not participate after being informed of their treatment assignment; 4 of the 6 withdrawals were assigned to haloperidol decanoate. Mean doses at week 48 were 493 mg/day of quetiapine (N = 16) and 170 mg/28 days of haloperidol decanoate (N = 9). Survival analysis showed no between-group differences in estimates of the number of patients remaining exacerbation-free over time. Both drugs were efficacious, but quetiapine was significantly better than haloperidol decanoate in controlling negative symptoms (p < .05). The incidence of extrapyramidal symptoms was low in both groups; patients receiving quetiapine showed significantly greater improvement in rigidity and akathisia (p < .05). CONCLUSION: Oral quetiapine was as efficacious as intramuscular haloperidol in preventing symptom exacerbation over 48 weeks in patients with schizophrenia or schizoaffective disorder, with fewer extrapyramidal symptoms, especially rigidity and akathisia. Quetiapine was more efficacious than haloperidol decanoate in treating negative symptoms. PMID- 15889953 TI - Dermatology precautions and slower titration yield low incidence of lamotrigine treatment-emergent rash. AB - OBJECTIVE: To assess treatment-emergent rash incidence when using dermatology precautions (limited antigen exposure) and slower titration during lamotrigine initiation. METHOD: We assessed rash incidence in 100 patients with DSM-IV bipolar disorder instructed, for their first 3 months taking lamotrigine, to avoid other new medicines and new foods, cosmetics, conditioners, deodorants, detergents, and fabric softeners, as well as sunburn and exposure to poison ivy/oak. Lamotrigine was not started within 2 weeks of a rash, viral syndrome, or vaccination. In addition, lamotrigine was titrated more slowly than in the prescribing information. Patients were monitored for rash and clinical phenomena using the Systematic Treatment Enhancement Program for Bipolar Disorder Clinical Monitoring Form. Descriptive statistics were compiled. RESULTS: No patient had serious rash. Benign rash occurred in 5 patients (5%) and resolved uneventfully in 3 patients discontinuing and 2 patients continuing lamotrigine. Two patients with rash were found to be not adherent to dermatology precautions. Therefore, among the remaining patients, only 3/98 (3.1%) had benign rashes. CONCLUSION: The observed rate of benign rash was lower than the 10% incidence in other clinical studies. The design of this study confounds efforts to determine the relative contributions of slower titration versus dermatology precautions to the low rate of rash. Systematic studies are needed to confirm these preliminary findings, which suggest that adhering to dermatology precautions with slower titration may yield a low incidence of rash with lamotrigine. PMID- 15889954 TI - Factors associated with medication adherence in African American and white patients with bipolar disorder. AB - BACKGROUND: African American patients may be less likely than white patients to adhere to maintenance pharmacotherapy for bipolar disorder. The purpose of this study was to examine rates of medication nonadherence, self-perceived reasons for nonadherence, and attitudes associated with non-adherence in these ethnic groups. METHOD: 20 African American and 30 white subjects with DSM-IV bipolar I disorder participated in this study. At a single follow-up visit with patients at least 4 months after their first hospitalization for acute mania, we assessed demographics, symptom severity, degrees of adherence, reasons for non-adherence, and self-perceptions regarding factors previously associated with nonadherence using a visual analog scale (VAS). The cross-sectional data that are the subject of this report were obtained from July 1, 2002, through June 30, 2004. RESULTS: Over 50% of participants in each group were currently either fully or partially nonadherent with medications. Greater than 20% of participants in each group denied having bipolar disorder and described physical side effects from medications as contributing to nonadherence. In principal components analysis of the VAS, 2 components were identified. The first component contained patient related factors associated with nonadherence, while the second contained a combination of illness- and medication-related factors. African American participants were more likely to endorse patient-related factors associated with nonadherence relative to white participants. Specifically, African Americans self endorsed a fear of becoming addicted to medications and feeling that medications were symbols of mental illness. CONCLUSION: Findings suggest that both African American and white patients with bipolar disorder demonstrate poor medication adherence that they attribute to illness/medication-related factors (denial of illness, physical side effects). However, patient-related factors (fear of addiction, medication as a symbol of illness) accounted for ethnic differences on self-perceived ratings of nonadherence factors. Differences in the reasons for nonadherence relative to culturally biased self-perceptions may help explain nonadherence behaviors in the African American community. PMID- 15889955 TI - Reading reports of clinical trial results. PMID- 15889956 TI - Use of olanzapine in anorexia nervosa. PMID- 15889958 TI - Injection site pain with long-acting risperidone. PMID- 15889959 TI - Do antihypertensives make tranylcypromine safer? Three case reports. PMID- 15889963 TI - The functional assessment of renal artery stenosis. PMID- 15889961 TI - Development of subtle psychotic symptoms with memantine: a case report. PMID- 15889965 TI - Telmisartan/hydrochlorothiazide: a new fixed dose combination. AB - Many patients with hypertension require two or more antihypertensive drugs with complementary mechanisms of action to lower their blood pressure and attain the therapeutic goals specified in internationally accepted guidelines. Yet, these latter guidelines offer the choice of fixed dose combinations as possible first step therapies. The angiotensin II type 1-receptor antagonist telmisartan and the diuretic hydrochlorothiazide are two antihypertensive agents that have a well recognized clinical efficacy. Their combination was shown in randomized, controlled trials to be more effective than each agent alone in lowering blood pressure, due to a dual and synergistic mechanism. Indeed, combining telmisartan with hydrochlorothiazide enhances the antihypertensive efficacy of telmisartan in almost two-thirds of hypertensive patients with mild-to-moderate hypertension. The pharmacokinetics and -dynamics of the combination are similar to monotherapy, and the addition of hydrochlorothiazide to telmisartan does not modify the excellent tolerability profile of the drug. PMID- 15889966 TI - Azimilide dihydrochloride. AB - Azimilide dihydrochloride is an antiarrhythmic drug with Vaughn Williams class III properties, which blocks both fast (IKr) and slow (IKs) components of the delayed rectifier cardiac potassium channel. The drug slows the heart rate slightly and, like other class III antiarrhythmic drugs, prolongs ventricular repolarization and thus, the QT interval. Unlike sotalol, another class III antiarrhythmic drug, azimilide does not exhibit reverse-use dependence, that is, its binding characteristics and effectiveness are not related to the heart rate. Azimilide is 85% bioavailable, reaches peak blood concentrations in 6-8 h and has a long elimination half-life of 114 h. Clinical trials have utilized once-daily dosing. These trials have tested the use of the drug for patients with supraventricular and ventricular arrhythmias. PMID- 15889967 TI - Adiposopathy, metabolic syndrome, quantum physics, general relativity, chaos and the Theory of Everything. AB - Excessive fat (adiposity) and dysfunctional fat (adiposopathy) constitute the most common worldwide epidemics of our time -- and perhaps of all time. Ongoing efforts to explain how the micro (adipocyte) and macro (body organ) biologic systems interact through function and dysfunction in promoting Type 2 diabetes mellitus, hypertension and dyslipidemia are not unlike the mechanistic and philosophical thinking processes involved in reconciling the micro (quantum physics) and macro (general relativity) theories in physics. Currently, the term metabolic syndrome refers to a constellation of consequences often associated with excess body fat and is an attempt to unify the associations known to exist between the four fundamental metabolic diseases of obesity, hyperglycemia (including Type 2 diabetes mellitus), hypertension and dyslipidemia. However, the association of adiposity with these metabolic disorders is not absolute and the metabolic syndrome does not describe underlying causality, nor does the metabolic syndrome necessarily reflect any reasonably related pathophysiologic process. Just as with quantum physics, general relativity and the four fundamental forces of the universe, the lack of an adequate unifying theory of micro causality and macro consequence is unsatisfying, and in medicine, impairs the development of agents that may globally improve both obesity and obesity-related metabolic disease. Emerging scientific and clinical evidence strongly supports the novel concept that it is not adiposity alone, but rather it is adiposopathy that is the underlying cause of most cases of Type 2 diabetes mellitus, hypertension and dyslipidemia. Adiposopathy is a plausible Theory of Everything for mankind's greatest metabolic epidemics. PMID- 15889968 TI - Acute stroke hypertension: current and future management. AB - The management of hypertension in acute stroke remains a hotly debated issue. Clinical practice varies widely between physicians, and both European and US guidelines reflect the uncertainty surrounding this question. Although there is a large amount of data that, on the whole, tends to support a connection between poststroke hypertension and hypotension and worse outcome, there have been few randomized controlled trials to clarify whether pharmacologic intervention is safe or beneficial. Data from secondary prevention trials convincingly demonstrate the benefits of controlling hypertension after a stroke but do not guide us as to how early to implement therapy. There is even less information from trials regarding the use of pressor agents in hypotensive stroke patients. This review discusses the dilemmas in the management of acute stroke hypertension and summarizes the available evidence from studies involving a variety of both depressor and pressor agents. The authors detail the ongoing studies that will help to answer some of the outstanding questions and summarizes the existing guidelines regarding indications for acute stroke blood pressure manipulation currently available to physicians. PMID- 15889969 TI - Renovascular hypertension. AB - Renal artery disease is the most common cause of potentially curable secondary hypertension, with atherosclerosis as the major cause of renal artery stenosis. Fibromuscular dysplasia is a less common cause of renal artery stenosis and is most frequently observed in premenopausal women. Renal artery stenosis is likely to be underappreciated and is more common in patients with other vascular disease (e.g., coronary or peripheral arterial disease). The diagnosis of renal artery stenosis requires a high clinical index of suspicion as well as an appropriate imaging strategy, with currently effective diagnostic modalities including magnetic resonance imaging, computed tomography and renal artery duplex ultrasonography. The current treatment of choice for atherosclerotic renal artery stenosis is balloon angioplasty and secondary stenting, whereas angioplasty alone is the treatment for renal artery stenosis secondary to fibromuscular dysplasia. Expected outcomes following revascularization include improved blood pressure control and possibly renal preservation. Ongoing studies will hopefully identify patient characteristics that will achieve the most benefit from percutaneous revascularization as well as the impact of percutaneous revascularization with drug-eluting stents and embolic protection devices. PMID- 15889970 TI - Acute aortic syndromes: aortic dissections, penetrating aortic ulcers and intramural aortic hematomas. AB - Acute aortic syndromes, including dissections, intramural hematomas and penetrating aortic ulcers, are a catastrophic clinical entity that are relatively uncommon. A high index of clinical suspicion along with proper imaging modalities are critical in making a prompt and accurate diagnosis for immediate management and to improve survival of the patient. PMID- 15889971 TI - Combined hypertension and orthostatic hypotension in older patients: a treatment dilemma for clinicians. AB - The combination of hypertension and orthostatic hypotension in older individuals is becoming increasingly recognized. Managing this combination of disorders presents a treatment dilemma -- how to lower blood pressure to provide cardiovascular risk protection without predisposing to syncope. At present, there is no specific evidence base available with regard to managing such patients. Some antihypertensive drug classes (e.g., alpha-blockers) appear more problematic in this regard than others. In the absence of controlled-trial evidence, use of antihypertensives with a more gradual onset of effect commenced at lower doses and use of lower-limb compression hosiery appears to be a reasonable approach. Abdominal compression devices and elevating the head of the bed may also help to combat orthostatic hypotenstion in older patients with hypertension and warrant future research. PMID- 15889973 TI - Dyslipidemia and lipid-lowering therapy in the elderly. AB - According to the current evidence, the fastest growing proportion of patient populations seeking healthcare is those over 65 years of age. Coronary artery disease and subclinical atherosclerosis are highly prevalent in this group of patients and are strongly linked to dyslipidemia, a well-established risk factor for atherosclerosis. Treating dyslipidemia in this group of patients requires specific knowledge and understanding of common dyslipidemias and the relative safety of various pharmacologic agents in the presence of possible multiple comorbidities. Lifestyle modification remains the first step in the treatment of dyslipidemia; however, it can be difficult to sustain and achieve acceptable compliance in the elderly and it is best used in combination with drug therapy. Statins are widely accepted as the first-line therapy. Several recent studies have demonstrated that statins are safe and effective in the elderly. However, it is important to note that there is very limited data regarding the effects of dyslipidemia treatment on morbidity and mortality in patients over 85 years of age. In summary, the clinicians must recognize that the presence of dyslipidemia in the elderly poses substantial risk of coronary events and stroke. The available evidence has demonstrated that in most elderly patients who are at increased risk for cardiovascular morbidity and mortality, treatment of dyslipidemia with appropriate therapy reduces the risk, and when used carefully with close monitoring for safety, the treatment is generally well tolerated. With increasing life expectancy, it is critical for physicians to recognize the importance of detection and treatment of dyslipidemia in the elderly. PMID- 15889972 TI - ROCKs as therapeutic targets in cardiovascular diseases. AB - There is growing evidence that Rho-kinases (ROCKs), the immediate downstream targets of the small guanosine triphosphate-binding protein Rho, may contribute to cardiovascular disease. ROCKs play a central role in diverse cellular functions such as smooth muscle contraction, stress fiber formation and cell migration and proliferation. Overactivity of ROCKs is observed in cerebral ischemia, coronary vasospasm, hypertension, vascular inflammation, arteriosclerosis and atherosclerosis. ROCKs, therefore, may be an important and still relatively unexplored therapeutic target in cardiovascular disease. Recent experimental and clinical studies using ROCK inhibitors such as Y-27632 and fasudil have revealed a critical role of ROCKs in embryonic development, inflammation and oncogenesis. This review will focus on the potential role of ROCKs in cellular functions and discuss the prospects of ROCK inhibitors as emerging therapy for cardiovascular diseases. PMID- 15889974 TI - Adiponectin: linking the metabolic syndrome to its cardiovascular consequences. AB - Obesity and its related disorders, glucose intolerance, hypertension and hyperlipidemia, collectively named the metabolic syndrome, result in substantial cardiovascular morbidity and mortality. Recent data point to several underlying regulatory mechanisms through which obesity links these various outcomes. Adipose tissue is now understood to function not merely as a passive energy storage depot but as an active endocrine organ, producing a variety of bioactive substances termed adipocytokines. Adiponectin, an adipocytokine first described as the most abundant protein produced by adipocytes, appears to serve as a central regulatory protein in many of the physiologic pathways controlling lipid and carbohydrate metabolism, and to mediate various vascular processes. Adiponectin displays both anti-inflammatory and antiatherogenic properties. Unlike other adipocytokines, its levels are paradoxically decreased in obesity and insulin-resistance states including metabolic syndrome and diabetes, as well as hypertension and coronary artery disease. This review will detail the relationship of adiponectin to various features of obesity and insulin-resistance syndromes, as well as its relationship to the cardiovascular complications of these disorders. PMID- 15889975 TI - Noninvasive cardiac imaging in the evaluation of suspected acute coronary syndromes. AB - Optimal management of patients presenting with chest pain to the emergency department is a major challenge, both in terms of a diagnostic dilemma and consumption of resources. The triage of such patients can be aided vastly by the appropriate use of noninvasive imaging. Noninvasive imaging modalities such as echocardiogram, radionuclide perfusion studies, positron emission tomography, cardiac magnetic resonance imaging and computed tomography have all been demonstrated to have favorable diagnostic and prognostic value, with an enhanced sensitivity to detect acute ischemia. A normal noninvasive evaluation in the appropriate clinical setting presents a strong argument against acute ischemia as an etiology of the chest pain. Randomized trials of both rest and stress imaging in the emergency department have confirmed a reduction in unnecessary hospitalizations and cost savings without compromising the safety of the patient. Cardiac magnetic resonance and computed tomography would provide an insight into subendocardial ischemia, the detection of which has previously been difficult, using single-photon emission tomography and echocardiography. In this review, novel hot-spot imaging modalities are discussed including infarct-avid imaging agents and ischemia-avid imaging agents, thus elucidating the pathophysiology of reperfusion-induced cell death. These agents represent work in evolution and are likely to be used routinely in the future as understanding of coronary syndromes and coronary artery disease becomes clearer. PMID- 15889976 TI - N-terminal proBNP: a novel biomarker for the diagnosis, risk stratification and management of congestive heart failure. AB - Congestive heart failure is a leading cause of morbidity and mortality throughout the world and is now the leading cause of hospitalizations in adults over 65 years of age with an estimated annual expenditure in excess of USD 20 billion. In addition, it is the only cardiovascular disorder that continues to increase in both incidence and prevalence, and as the population continues to age, it is expected that the prevalence of this disease will continue to rise. Ironically, the armamentarium of medications that decrease mortality due to congestive heart failure also continues to grow; however, the relative number of eligible patients with congestive heart failure (or at risk for congestive heart failure) that receive these important therapies remains low. Thus, better tools to aid the early diagnosis and management of this disease are needed. Testing for natriuretic peptide markers, such as B-type natriuretic peptide or its amino terminal fragment, has emerged as an important tool to assist in the optimal diagnosis and risk stratification of patients with congestive heart failure and may also play a valuable role in guiding therapy. PMID- 15889977 TI - Adverse cardiorenal effects of aldosterone: is aldosterone antagonism beneficial? AB - Aldosterone has recently been recognized as an important factor in the development and progression of cardiorenal disease. Animal and human data suggest that aldosterone contributes importantly to several disease states. These include congestive heart failure, coronary heart disease and progression of kidney disease. Recently, the discovery that aldosterone antagonists decrease pathologic injury in the kidneys and nonepithelial tissues, such as the myocardium and endothelium, has generated great controversy regarding the actual mechanisms of benefit of these agents. The available data is reviewed and conclusions drawn regarding the relative benefits of modulating aldosterone effects in the cardiovascular system and the kidney. In particular, the authors review their effects on reductions in cardiovascular events and progression of chronic kidney disease, as well as the safety and tolerability of these agents. PMID- 15889978 TI - Hepatocyte growth factor as potential cardiovascular therapy. AB - Hepatocyte growth factor is a mesenchyme-derived pleiotropic factor that regulates the growth, motility and morphogenesis of various types of cells, and is also a member of the angiogenic growth factors. Hepatocyte growth factor is secreted by vascular endothelial cells and smooth muscle cells, and the hepatocyte growth factor receptor, c-met, was also observed in these vascular cells. Treatment of human aortic endothelial cells with recombinant hepatocyte growth factor resulted in a significant increase in cell proliferation, accompanied by mitogen-activated protein kinase and Akt/protein kinase B phosphorylation. Recently, a novel therapeutic strategy for ischemic diseases using angiogenic growth factors to augment collateral artery development has been proposed. As preclinical study of gene therapy using hepatocyte growth factor to treat peripheral arterial disease, naked hepatocyte growth factor plasmid was intramuscularly injected into the ischemic hind limb of rabbits in order to evaluate its angiogenic activity. Intramuscular injection of hepatocyte growth factor plasmid once on day 10 following surgery, produced significant augmentation of collateral vessel development in the ischemic limb on day 30. In the clinical setting, the authors further investigated the safety and efficacy of hepatocyte growth factor plasmid DNA in patients with critical limb ischemia, in a prospective open-labeled trial. Intramuscular injection of naked plasmid DNA was performed in the ischemic limbs of six patients with critical limb ischemia with arteriosclerosis obliterans (n = 3) or Buerger disease (n = 3) graded as Fontaine III or IV. In the efficacy evaluation, a reduction of pain scale of more than 1 cm on a visual analog pain scale was observed in five out of six patients. An increase in ankle pressure index of more than 0.1 was observed in five out of five patients. The long diameter of eight out of 11 ischemic ulcers in four patients was reduced by more than 25%. Intramuscular injection of naked hepatocyte growth factor plasmid is safe, feasible and can achieve successful improvement of ischemic limbs. Although the present data were obtained to demonstrate safety in a Phase I/early Phase II trial, the initial clinical outcome with hepatocyte growth factor gene transfer seems to indicate its usefulness as sole therapy for critical limb ischemia. Randomized placebo controlled clinical trials of alternative dosing regimens of gene therapy will be required to define the efficiency of this therapy. PMID- 15889979 TI - Angiogenic therapy for coronary artery and peripheral arterial disease. AB - Neovascularization in chronically ischemic adult cardiac and skeletal muscle results from the processes of angiogenesis, arteriogenesis and vasculogenesis. Therapeutic angiogenesis describes an emerging field of cardiovascular medicine whereby new blood vessels are induced to grow to supply oxygen and nutrients to cardiac or skeletal muscle rendered ischemic as a result of progressive atherosclerosis. Various techniques have been utilized to promote new blood vessel growth in the heart and extremities, including mechanical means such as surgical or percutaneous myocardial laser revascularization, angiogenic growth factor therapies involving members of the vascular endothelial growth factor and fibroblast growth factor families, and more recently, cellular-based therapies using stem cells known as endothelial progenitor cells or angioblasts. The following review discusses each of these treatment strategies in detail including both preclinical and clinical data for their use in peripheral arterial and coronary artery disease. PMID- 15889981 TI - Smallpox vaccines: circling the wagons. PMID- 15889984 TI - 8th annual meeting of the Regional Cancer Center Consortium. 26-28 February, 2005. Penn State Cancer Center, Hershey, PA, USA. PMID- 15889985 TI - PEDIARIX: clinical trials. AB - PEDIARIX is the first pentavalent combination vaccine licensed for use in infants in the USA. This vaccine is indicated for the prevention of diphtheria, tetanus, pertussis, hepatitis B and poliovirus. This article reviews the available data regarding the vaccine's immunogenicity and safety. PMID- 15889986 TI - Paving the way for the introduction of new vaccines into developing countries. AB - The great advances in vaccine R&D over the last 20 years have not been paralleled by the regular introduction of new vaccines into the market thus far. This situation acquires more dramatic proportions in developing countries, where preventable infectious diseases still impose a major public health problem. Furthermore, most of the new developments are being conceived to target the market of industrialized countries and it is foreseeable that their introduction in low-to-middle income countries will be difficult if at any time possible. Strengthening their own capacities for R&D and production is likely to be the most reasonable avenue to ensure that new vaccines will become a sustainable reality for developing countries. Concerted efforts that draw together local capacities (industry and academy) with the experience of large global manufacturers, could have a major impact and provide a great example of an effective partnership to achieve this. PMID- 15889987 TI - Are we ready for pandemic influenza H5N1? AB - Influenza is an important annual respiratory pathogen with the potential to cause infrequent pandemics with devastating consequences. The establishment of highly pathogenic avian influenza H5N1 as an endemic virus within duck and poultry populations in Asia increases the possibility of adaptation to humans and the threat of an emerging pandemic. Vaccines are the mainstay of prophylaxis against influenza, but there are technical and safety issues that must be overcome in the development of vaccines in order to combat avian influenza. Pandemic preparedness plans have been developed by national and international authorities but may be compromised by a lack of readily available interventions. PMID- 15889990 TI - ELISA as a possible alternative to the neutralization test for evaluating the immune response to poliovirus vaccines. AB - This review describes several enzyme-linked immunosorbent assay (ELISA) techniques proposed to replace the neutralization test for detecting neutralization-relevant antibodies to polioviruses in recipients of inactivated poliovirus vaccine and oral poliovirus vaccine, and for seroepidemiologic studies. Comparisons of results from ELISA and the neutralization test suggest that ELISA variants, based on the principle of blocking or binding inhibition that emulate the neutralization test, might offer an alternative to the neutralization test. However, to replace the neutralization test with ELISA would first require extensive studies with very large numbers of serum samples, including sera having low titers of neutralizing antibodies, in order to obtain reliable and statistically sound validation. PMID- 15889991 TI - Acellular pertussis vaccines in Japan: past, present and future. AB - An antivaccine movement developed in Japan as a consequence of increasing numbers of adverse reactions to whole-cell pertussis vaccines in the mid-1970s. After two infants died within 24 h of the vaccination from 1974 to 1975, the Japanese government temporarily suspended vaccinations. Subsequently, the public and the government witnessed the re-emergence of whooping cough, with 41 deaths in 1979. This series of unfortunate events revealed to the public that the vaccine had, in fact, been beneficial. Furthermore, researchers and the Japanese government proceeded to develop safer pertussis vaccines. Japan now has the most experience worldwide with acellular pertussis vaccines, being the first country to have approved their use. This review describes the major events associated with the Japanese vaccination program. The Japanese experience should be valuable to other countries that are considering the development and use of such vaccines. PMID- 15889992 TI - Nanoparticles as carriers for nasal vaccine delivery. AB - At present, there is considerable excitement within the nanotechnology field with regard to the potential use of nanosystems as carriers for mucosal vaccine delivery. Indeed, many of the vaccines available, including protein antigens and DNA vaccines, are very unstable and need to be protected from degradation in the biologic environment. In addition, their efficacy is limited by their poor capacity to cross biologic barriers and reach the target sites. As a consequence, the design of appropriate antigen carriers that could help overcome these problems has become a significant challenge. The goal of the present article is to review the recent advances in the design of polymeric nanosystems intended to be used as carriers for nasal vaccine delivery. More specifically, the authors present nanocarriers that have been made of safe materials, such as biodegradable polyesters and polysaccharides. The information accumulated regarding the in vivo behavior of these nanocarriers indicates that they are able to facilitate the transport of the associated antigen across the nasal epithelium, thus leading to efficient antigen presentation to the immune system. Furthermore, the results suggest that not only the size and surface properties but also the polymer composition and the structural architecture of the nanosystems are critical for the optimization of these antigen carriers. In conclusion, future studies intended to provide increased knowledge regarding these properties and how they relate to the efficiency of the immune responses, will undoubtedly affect the design of new and more effective nasal vaccine delivery strategies. PMID- 15889993 TI - Cancer risk associated with receipt of vaccines contaminated with simian virus 40: epidemiologic research. AB - Simian virus (SV)40 was an accidental contaminant of poliovirus vaccines used widely in the USA and other countries in 1955-1962. Exposure to SV40 via contaminated vaccines has led to concern as SV40 causes cancer in laboratory animals. In addition, some laboratories, although not all, have detected SV40 DNA in human tumors including mesothelioma, certain brain tumors, osteosarcoma and non-Hodgkin's lymphoma. This article reviews the data regarding contamination of poliovirus vaccines with SV40 and summarizes the results from epidemiologic studies of vaccine recipients. Long-term follow-up studies have not revealed recipients of SV40-contaminated poliovirus vaccines to be at an increased risk for cancer. Thus, these studies are somewhat reassuring and indicate that either SV40 does not readily infect humans or, following infection, does not cause cancer. Recognizing that the history of SV40 contamination of vaccines highlights an inherent risk of contamination of vaccines with adventitious agents, the Institute of Medicine recently called for the development of a comprehensive US plan to prevent vaccine contamination and respond to potential contamination events when they arise. PMID- 15889994 TI - Vaccination against group B streptococcus. AB - Streptococcus agalactiae (Group B streptococcus) is an important cause of disease in infants, pregnant women, the elderly and in immunosuppressed adults. An effective vaccine is likely to prevent the majority of infant disease (both early and late onset), as well as Group B streptococcus-related stillbirths and prematurity, to avoid the current real and theoretical limitations of intrapartum antibiotic prophylaxis, and to be cost effective. The optimal time to administer such a vaccine would be in the third trimester of pregnancy. The main limitations on the production of a Group B streptococcus vaccine are not technical or scientific, but regulatory and legal. A number of candidates including capsular conjugate vaccines using traditional carrier proteins such as tetanus toxoid and mutant diphtheria toxin CRM197, as well as Group B streptococcus-specific proteins such as C5a peptidase, protein vaccines using one or more Group B streptococcus surface proteins and mucosal vaccines, have the potential to be successful vaccines. The capsular conjugate vaccines using tetanus and CRM197 carrier proteins are the most advanced candidates, having already completed Phase II human studies including use in the target population of pregnant women (tetanus toxoid conjugate), however, no definitive protein conjugates have yet been trialed. However, unless the regulatory environment is changed specifically to allow the development of a Group B streptococcus vaccine, it is unlikely that one will ever reach the market. PMID- 15889995 TI - Reimmunization after hematopoietic stem cell transplantation. AB - Hematopoietic stem cell transplant recipients lose immune memory of exposure to infectious agents and vaccines accumulated through a lifetime, and therefore need to be revaccinated. Reimmunization protocols vary greatly among hematopoietic stem cell transplant centers. Diphtheria and tetanus toxoids, pertussis vaccine, Haemophilus influenza type B conjugate, 23-valent pneumococcal polysaccharide, inactivated influenza and polio vaccine and live attenuated measles-mumps-rubella vaccine are the currently recommended vaccines to be included in a vaccination program after hematopoietic stem cell transplant. Other variables, such as stem cell source, new adjuvants, T-cell depleted transplants, nonmyeloablative conditioning and donor immunization have recently been introduced and a constant update of current recommendations are needed. Studies recently published, the use of other vaccines and the perspectives for different vaccination protocols are discussed in this review. PMID- 15889996 TI - Ricin: current understanding and prospects for an antiricin vaccine. AB - Ricin is a potent cytotoxin that can be rapidly internalized into mammalian cells leading to cell death. The ease in obtaining the toxin and its deadly nature combine to implicate ricin as a convenient agent for bioterrorism. Research into the mechanism of toxicity, as well as strategies for treatment and protection from the toxin has been widely undertaken for a number of years. This article reviews the current understanding of the mechanism of action of the toxin, the clinical effects of ricin intoxication and how these relate to current and continuing prospects for vaccine development. PMID- 15889997 TI - Oxidative modification of proteins in cell signaling. PMID- 15889998 TI - Hyperglycemia potentiates H(2)O(2) production in adipocytes and enhances insulin signal transduction: potential role for oxidative inhibition of thiol-sensitive protein-tyrosine phosphatases. AB - Insulin signal transduction in adipocytes is accompanied by a burst of cellular hydrogen peroxide (H(2)O(2)) that facilitates insulin signaling by inhibiting thiol-dependent protein-tyrosine phosphatases (PTPs) that are negative regulators of insulin action. As hyperglycemia is associated with increased cellular reactive oxygen species, we postulated that high glucose conditions might potentiate the H(2)O(2) generated by insulin and modulate insulin-stimulated protein phosphorylation. Basal H(2)O(2) generation was increased threefold in differentiated 3T3-L1 adipocytes by growth in 25 mM glucose versus 5 mM glucose. High glucose increased the sensitivity of the insulin-stimulated H(2)O(2) signal to lower concentrations of insulin. Basal endogenous total PTP activity and the activity of PTP1B, a PTP implicated in the negative regulation of insulin signaling, were reduced in high glucose conditions, and their further reduction by insulin stimulation was more enhanced in high versus low glucose medium. Phosphorylation of the insulin receptor, IRS-1, and Akt in response to insulin was also significantly enhanced in high glucose conditions, especially at submaximal insulin concentrations. In primary rat adipocytes, high glucose increased insulin-stimulated H(2)O(2) production and potentiated the oxidative inhibition of total PTP and PTP1B activity; however, insulin signaling was not enhanced in the primary cells in high glucose apparently due to cross-regulation of insulin-stimulated protein phosphorylation by activation of protein kinase C (PKC). These studies indicate that high glucose can enhance insulin stimulated H(2)O(2) generation and augment oxidative PTP inhibition in cultured and primary adipocytes, but the overall balance of insulin signal transduction is determined by additional signal effects in high glucose, including the activation of PKC. PMID- 15889999 TI - Oxidized and ubiquitinated proteins may predict recovery of postischemic cardiac function: essential role of the proteasome. AB - This study examined the hypothesis that postischemic levels of oxidized and/or ubiquitinated proteins may be predictive of functional recovery as they may be indicative of activity of the 20S and/or 26S proteasomes, respectively. Subjecting isolated rat hearts to 15 min of ischemia had no effect on 20S- and 26S-proteasome activities; however, both were significantly (p < 0.05) decreased by 70% and 54%, respectively, following 30 min of ischemia and 60 min of reperfusion, changes associated with increased levels of protein carbonyls and ubiquitinated proteins. Preischemic treatment of hearts with the proteasome inhibitor, MG132, resulted in dose-dependent decreases (p < 0.05) in recovery of postischemic function [MG132 (microM), heart rate x pressure product: 0, 11,158 +/- 2,423; 6, 11,400 +/- 3,009; 12, 5,513 +/- 2,225; 25, 2,325 +/- 992] and increased accumulation of ubiquitinated proteins. Preconditioning with repetitive ischemia (IP) or preischemic treatment with nicorandil (Nic) resulted in a significant increase in postischemic 20S-proteasome activity after 60 min of reperfusion (control, 95 +/- 4; IP, 301 +/- 65; Nic, 242 +/- 61 fluorescence units). Only Nic had similar effects on 26S-proteasome activity. These results support the conclusion that a correlation exists between eventual recovery of postischemic function and levels of oxidized and/or ubiquitinated proteins, a phenomenon that may be dependent on activity of the 20S and 26S proteasomes. PMID- 15890000 TI - Homocysteine and redox signaling. AB - Homocysteine is a thiol-containing amino acid that has gained notoriety because its elevation in the plasma is correlated with complex and multifactorial diseases, including cardiovascular diseases, neurodegenerative diseases, and neural tube defects. Homocysteine is redox-active, and its toxic effects have been frequently attributed to direct or indirect perturbation of redox homeostasis. Although the literature on the pathophysiology of elevated homocysteine is rather extensive, a very wide range of concentrations of this amino acid has been used in these studies ranging from normal to pathophysiological to unphysiological. It is clear that homocysteine induces varied responses that are specific to cell type and that cells, depending on their origin, display a wide range of sensitivity to homocysteine. In this review, we focus on the redox signaling pathways that have been connected to homocysteine in vascular (endothelial and smooth muscle) cells and in neuronal cells. We also discuss redox regulation of the key enzymes involved in homocysteine clearance: methionine synthase, betaine-homocysteine methyltranferase, and cystathionine beta-synthase. PMID- 15890001 TI - Functions and mechanisms of redox regulation of cysteine-based phosphatases. AB - Reactive oxygen species (ROS) have been implicated as mediators of cell-signaling responses, particularly in pathways involving protein tyrosine phosphorylation. One mechanism by which ROS are thought to exert their effects is through the reversible regulation of cysteine-based phosphatases (CBPs). The CBPs, which include protein tyrosine phosphatases (PTPs), dual-specificity phosphatases, low molecular-weight PTPs, and the lipid phosphatase PTEN, all contain a nucleophilic catalytic cysteine within a conserved motif that enables these enzymes to dephosphorylate phosphoproteins or phospholipids. In addition to enabling phosphatase activity, the nucleophilic catalytic cysteines of CBPs are also highly susceptible to oxidation, a property that permits redox regulation of this enzyme family. In this review, we discuss the evidence implicating ROS as mediators of CBP activity within signaling pathways and discuss how specificity of ROS-dependent signaling involving CBPs may be achieved. We also discuss the molecular mechanisms that facilitate the stabilization of a reversibly oxidized form of the catalytic cysteine. These mechanisms include the formation of disulfide bonds or the formation of a sulfenamide bond, a novel mechanism that was identified for PTP1B. Formation of either type of covalent bond may be accompanied by dramatic structural rearrangements that can affect downstream signaling events and allow for multitiered enzyme regulation. PMID- 15890002 TI - Anchorage-dependent cell growth: tyrosine kinases and phosphatases meet redox regulation. AB - Recent data have provided new insight concerning the regulation of nontransformed cell proliferation in response to both soluble growth factors and adhesive cues. Nontransformed cells are anchorage-dependent for the execution of the complete mitotic program and cannot avoid the concomitant signals starting from mitogenic molecules, as growth factors, and adhesive agents belonging to the extracellular matrix. Protein tyrosine kinases (PTKs) and phosphotyrosine phosphatases (PTPs) together with soluble small molecules have been included among intracellular signal transducers of growth factor and extracellular matrix receptors. Reactive oxygen species retain a key role during both growth factor and integrin receptor signaling, and these second messengers are recognized to be a synergistic point of confluence for anchorage-dependent growth signaling. Redox-regulated proteins include PTPs and PTKs, although with opposite regulation of enzymatic activity. Transient oxidation of PTPs leads to their inactivation, through the formation of an intramolecular S-S bridge. Conversely, oxidation of PTKs leads to their activation, either by direct SH modification or, indirectly, by concomitant inhibition of PTPs that leads to sustained activation of PTKs. This review will focus on the redox regulation of PTPs and PTKs during anchorage-dependent cell growth and its implications for tumor biology. PMID- 15890003 TI - Chemical considerations and biological selectivity of protein nitrosation: implications for NO-mediated signal transduction. AB - Nitric oxide (NO) is a diatomic free radical that plays an important role in the homeostatic regulation of the central nervous, immune, and cardiovascular systems. In addition to its interaction with guanylate cyclase, which results in the production of the second messenger cyclic GMP, there is now a large body of literature indicating that many of the effects associated with the production of NO are due to the nitrosation of cysteine residues in proteins. In this review, we outline the primary chemical pathways that may account for protein nitrosation in cells and tissues. The functional implications of protein nitrosation are discussed by using the p21(ras) subfamily of small monomeric GTPases and the cysteine-containing aspartate-specific proteases (caspases) as prototypical examples. Overall, in addition to the well characterized NO/O(2) reaction, there may exist multiple pathways accounting for protein nitrosation in cells. These include acid- and free radical-mediated mechanisms. Although protein nitrosation may not be limited to cysteine residues, there is now ample evidence that nitrosation reactions, in a fashion similar to oxidative modifications, may modulate the structure, activity, association, and localization of a specific subset of proteins in cells and tissues. PMID- 15890004 TI - Redox regulation and modification of proteins controlling chloroplast gene expression. AB - Chloroplasts are typical organelles of plant cells and represent the site of photosynthesis. As one very remarkable feature, they possess their own genome and a complete machinery to express the genetic information in it. The plastid gene expression machinery is a unique assembly of prokaryotic-, eukaryotic-, and phage like components because chloroplasts acquired a great number of regulatory proteins during evolution. Such proteins can be found at all levels of gene expression. They significantly expand the functional and especially the regulatory properties of the "old" gene expression system that chloroplasts inherited from their prokaryotic ancestors. Recent results show that photosynthesis has a strong regulatory effect on plastid gene expression. The redox states of electron transport components, redox-active molecules coupled to photosynthesis, and pools of reactive oxygen species act as redox signals. They provide a functional feedback control, which couples the expression of chloroplast genes to the actual function of photosynthesis and, by this means, helps to acclimate the photosynthetic process to environmental cues. The redox signals are mediated by various specific signaling pathways that involve many of the "new" regulatory proteins. Chloroplasts therefore are an ideal model to study redox-regulated mechanisms in gene expression control. Because of the multiple origins of the expression machinery, these observations are of great relevance for many other biological systems. PMID- 15890005 TI - Controlled elimination of intracellular H(2)O(2): regulation of peroxiredoxin, catalase, and glutathione peroxidase via post-translational modification. AB - The predominant enzymes responsible for elimination of hydrogen peroxide (H(2)O(2)) in cells are peroxiredoxins (Prxs), catalase, and glutathione peroxidases (GPxs). Evidence suggests that catalytic activities of certain isoforms of these H(2)O(2)-eliminating enzymes are extensively regulated via posttranslational modification. Prx I and Prx II become inactivated when phosphorylated on Thr(90) by cyclin B-dependent kinase Cdc2. In addition, the active-site cysteine of Prx I-IV undergoes a reversible sulfinylation (oxidation to cysteine sulfinic acid) in cells. Desulfinylation (reduction to cysteine) is achieved by a novel enzyme named sulfiredoxin. c-Abl and Arg nonreceptor protein tyrosine kinases associate with catalase in cells treated with H(2)O(2) by mechanisms involving the SH3 domains of the kinases and the Pro(293)PheAsnPro motif of catalase and activate catalase by phosphorylating it on Tyr(231) and Tyr(386). Similarily, GPx1 is activated by c-Abl- and Arg-mediated phosphorylation. The tyrosine phosphorylation is critical for ubiquitination dependent degradation of catalase. PMID- 15890006 TI - Oxidative damage in Parkinson's disease. PMID- 15890007 TI - The effect of endogenous dopamine in rotenone-induced toxicity in PC12 cells. AB - Deficiencies in Complex I have been observed in Parkinson's disease (PD) patients. Systemic exposure to rotenone, a Complex I inhibitor, has been shown to lead to selective dopaminergic cell death in vivo and toxicity in many in vitro models, including dopaminergic cell cultures. However, it remains unclear why rotenone seems to affect dopaminergic cells more adversely. Therefore, the role of dopamine (DA) in rotenone-induced PC12 cell toxicity was examined. Rotenone (1.0 muM) caused significant toxicity in differentiated PC12 cells, which was accompanied by decreases in ATP levels, changes in catechol levels, and increased DA oxidation. To determine whether endogenous DA makes PC12 cells more susceptible to rotenone, cells were treated with the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine (AMPT) to reduce DA levels prior to rotenone exposure, and then cell viability was measured. No changes in rotenone-induced toxicity were observed with or without AMPT treatment. However, a potentiation of toxicity was observed following coexposure of PC12 cells to rotenone and methamphetamine. To determine whether this effect was due to DA, PC12 cells were depleted of DA prior to methamphetamine and rotenone cotreatment, resulting in a large attenuation in toxicity. These findings suggest that DA plays a role in rotenone-induced toxicity and possibly the vulnerability of DA neurons in PD. PMID- 15890008 TI - Microarray expression profiling identifies early signaling transcripts associated with 6-OHDA-induced dopaminergic cell death. AB - The parkinsonian mimetic 6-hydroxydopamine (6-OHDA) has been shown to cause transcriptional changes associated with cellular stress and the unfolded protein response. As these cellular sequelae depend on upstream signaling events, the present study used functional genomics and proteomic approaches to aid in deciphering toxin-mediated regulatory pathways. Microarray analysis of RNA collected from multiple time points following 6-OHDA treatment was combined with data mining and clustering techniques to identify distinct functional subgroups of genes. Notably, stress-induced transcription factors such as ATF3, ATF4, CHOP, and C/EBP beta were robustly up-regulated, yet exhibited unique kinetic patterns. Genes involved in the synthesis and modification of proteins (various tRNA synthetases), protein degradation (e.g., ubiquitin, Herpud1, Sqstm1), and oxidative stress (Hmox1, Por) could be subgrouped into distinct kinetic profiles as well. Realtime PCR and/or two-dimensional electrophoresis combined with western blotting validated data derived from microarray analyses. Taken together, these data support the notion that oxidative stress and protein dysfunction play a role in Parkinson's disease, as well as provide a time course for many of the molecular events associated with 6-OHDA neurotoxicity. PMID- 15890009 TI - Toxicity of redox cycling pesticides in primary mesencephalic cultures. AB - A loss of nigrostriatal dopaminergic neurons is the primary neurodegenerative feature of Parkinson's disease. Paraquat, a known redox cycling herbicide, has recently been shown to kill selectively nigrostriatal dopaminergic cells in the mouse model. The purpose of this study was to test the ability of paraquat and other redox cycling pesticides to damage dopaminergic neurons in primary mesencephalic cultures. Addition of paraquat, diquat, or benzyl viologen to mesencephalic cultures induced morphological changes (e.g., dystrophic neuronal processes) consistent with dopaminergic cell injury. The three pesticides also caused cell death as assessed by a reduction of the number of tyrosine hydroxylase-immunoreactive neurons and a dose-dependent decrease in [(3)H]dopamine uptake. Quite interestingly, diquat and benzyl viologen were significantly more toxic than paraquat, probably reflecting their more pronounced ability to trigger redox cycling reactions. The data support a role of redox cycling as a mechanism of dopaminergic cell degeneration and suggest that the property of redox cycling should be taken into consideration when evaluating putative environmental risk factors for Parkinson's disease. PMID- 15890010 TI - The role of microglia in paraquat-induced dopaminergic neurotoxicity. AB - The herbicide paraquat (PQ) has been implicated as a potential risk factor for the development of Parkinson's disease. In this study, PQ (0.5-1 microM) was shown to be selectively toxic to dopaminergic (DA) neurons through the activation of microglial NADPH oxidase and the generation of superoxide. Neuron-glia cultures exposed to PQ exhibited a decrease in DA uptake and a decline in the number of tyrosine hydroxylase-immunoreactive cells. The selectivity of PQ for DA neurons was confirmed when PQ failed to alter gamma-aminobutyric acid uptake in neuron-glia cultures. Microglia-depleted cultures exposed to 1 microM PQ failed to demonstrate a reduction in DA uptake, identifying microglia as the critical cell type mediating PQ neurotoxicity. Neuron-glia cultures treated with PQ failed to generate tumor necrosis factor-alpha and nitric oxide. However, microglia enriched cultures exposed to PQ produced extracellular superoxide, supporting the notion that microglia are a source of PQ-derived oxidative stress. Neuron-glia cultures from NADPH oxidase-deficient (PHOX-/-) mice, which lack the functional catalytic subunit of NADPH oxidase and are unable to produce the respiratory burst, failed to show neurotoxicity in response to PQ, in contrast to PHOX+/+ mice. Here we report a novel mechanism of PQinduced oxidative stress, where at lower doses, the indirect insult generated from microglial NADPH oxidase is the essential factor mediating DA neurotoxicity. PMID- 15890011 TI - Energy status, ubiquitin proteasomal function, and oxidative stress during chronic and acute complex I inhibition with rotenone in mesencephalic cultures. AB - Complex I impairment with rotenone produces damage though a mechanism thought to be distinct from effects on mitochondrial respiration. The outcome of chronic rotenone on energy status in relation to toxicity, however, is unknown. To examine this, mesencephalic cultures were exposed to chronic, low-dose rotenone (5-100 nM, 8 days in vitro) or acute, high-dose rotenone (500 nM, 1-24 h), and ATP/ADP levels and toxicity were measured. Chronic exposure to 5-50 nM rotenone produced selective dopamine cell loss. High-dose rotenone produced nonselective damage at all exposure times. Chronic, low-dose rotenone (37.5 nM) decreased ATP/ADP gradually over several days to 40% of controls, whereas high-dose rotenone (500 nM, 1-6 h), collapsed ATP/ADP by 1 h of exposure. The ubiquitin proteasomal pathway, an ATP-dependent pathway, is implicated in Parkinson's disease and, thus, various rotenone exposures were examined for effects on ubiquitin proteasomal function. Chronic, low-dose rotenone (25-50 nM, 8 days), but not acute, high-dose rotenone (500 nM, 1-6 h), caused accumulation of ubiquitinated proteins, E1-ubiquitin activation, and increased proteasomal activities prior to toxicity even though both exposures increased free radical production. Findings show that selective dopamine cell loss and alterations in ubiquitin proteasomal function only occur with rotenone exposures that partially maintain ATP/ADP. High concentrations of rotenone that collapse energy status kill neurons in a nonselective manner independent of the ubiquitin proteasomal pathway. PMID- 15890012 TI - Role of oxidative damage in protein aggregation associated with Parkinson's disease and related disorders. AB - Parkinson's disease, the most common movement disorder, is characterized by the loss of brainstem neurons, specifically dopaminergic neurons in the substantia nigra, as well as the accumulation of neuronal cytoplasmic filamentous proteinaceous inclusions comprised of polymerized alpha-synuclein. It was reported recently that alpha-synuclein can induce the formation of filamentous tau inclusions, which are characteristic of disorders like Alzheimer's disease and Lewy body variant of Alzheimer's disease, suggesting that a similar mechanism may exist between alpha-synuclein fibrillogenesis and tau polymerization. Pathological brain inclusions comprised of alpha-synuclein or tau proteins are associated with a spectrum of neurodegenerative disorders, and oxidative and nitrative injury has been implicated in all of these diseases. However, the role of oxidative damage in alpha-synuclein and tau polymerization and pathological inclusion formation is complex. Differences in the level, type, and temporal sequence of the oxidative alterations appear to result in both inhibitory and stimulatory effects on the fibrillogenesis of these proteins. PMID- 15890013 TI - Reactive oxygen and nitrogen species: weapons of neuronal destruction in models of Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disease whose etiology and pathogenesis remain mainly unknown. To investigate its cause and, more particularly, its mechanism of neuronal death, numerous in vivo experimental models have been developed. Currently, both genetic and toxic models of PD are available, but the use of neurotoxins such as 6-hydroxydopamine, paraquat, 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine, and rotenone are still the most popular means for modeling the destruction of the nigrostriatal dopaminergic neurons seen in PD. These four neurotoxins, although distinct in their intimate cytotoxic mechanisms, kill dopaminergic neurons via a cascade of deleterious events that consistently involves oxidative stress. Herein, we review and compare the molecular mechanisms of 6-hydroxydopamine, paraquat, 1-methyl-4-phenyl 1,2,3,6- tetrahydropyridine, and rotenone, placing the emphasis of our discussion on how reactive oxygen and nitrogen species contribute to the neurotoxic properties of these four molecules. As the reader will discover, to achieve the above stated goal, we had to not only appraise recent findings, but also revisit earlier landmark studies to provide a comprehensive view on this topic. This approach also enabled us to describe how our understanding of the mechanism of actions of certain toxins has evolved over time, which is particularly striking in the case of the quatrogenarian neurotoxin, 6-hydroxydopamine. PMID- 15890014 TI - Antioxidants as therapy for Parkinson's disease. PMID- 15890015 TI - Redox control of the cell cycle: a radical encounter. PMID- 15890016 TI - Carbon monoxide signaling in promoting angiogenesis in human microvessel endothelial cells. AB - Heme oxygenase isoforms (HO-1/HO-2) catalyze the conversion of heme to carbon monoxide (CO) and bilirubin. In this study, HO-1-deficient endothelial cells were transduced with HO-1 in the antisense orientation to determine whether supplementation with CO or bilirubin would regulate cell proliferation and angiogenesis. Western blotting, enzyme activity, CO and prostaglandin E(2) (PGE(2)) production, and cell-cycle analysis were used to assess transgenic expression and functionality of the recombinant protein. A Matrigel matrix was used for assessment of in vitro capillary formation. Transduction with HO-1 antisense resulted in decreased capillary formation, cell proliferation, and cell cycle progression, and increased PGE(2) production compared with control. HO-1 deficiency was also associated with increased expression of p21 and p27, but had no significant effect on p16 and p53. We also compared two different CO donors for their ability to rescue angiogenesis. Compared with control, HO-1-deficient endothelial cells showed increased angiogenesis following tricarbonyldichlororuthenium( II) dimer ([Ru(CO)(3)Cl(2)](2)) (CORM-1) starting at 50 microM, whereas tricarbonylchloro(glycinato) ruthenium(II) (CORM-3), starting at 25 microM, was a potent enhancer of angiogenesis. The addition of bilirubin did not restore angiogenesis. These data suggest that HO-mediated angiogenesis and cell proliferation were dependent on HO-1- and not HO-2-derived CO. PMID- 15890017 TI - Differential susceptibility of nonmalignant human breast epithelial cells and breast cancer cells to thiol antioxidant-induced G(1)-delay. AB - Reactive oxygen species (ROS) and ROS signaling have been implicated in a variety of human pathophysiological conditions that involve aberrant cellular proliferation, particularly cancer. We hypothesize that intracellular redox state differentially affects cell-cycle progression in nonmalignant versus malignant cells. The thiol antioxidant, N-acetyl-L-cysteine (NAC), was used to alter intracellular redox state in nonmalignant human breast epithelial (MCF-10A) and breast cancer cells (MCF-7 and MDA-MB-231). Treatment of cells with NAC resulted in significant augmentation of intracellular small-molecular-weight thiols, glutathione and cysteine. In addition, NAC treatment decreased oxidation of a prooxidant-sensitive dye in MCF-10A cells, but not in MDA-MB-231 and MCF-7 cells. NAC-induced shifts in intracellular redox state toward a more reducing environment caused G(1) delays in MCF-10A cells without causing any significant changes in MCF-7 and MDA-MB-231 cell-cycle progression. NAC treatment of MCF-10A (but not MCF-7 and MDA-MB-231) was accompanied by a decrease in cyclin D1 and an increase in p27 protein levels, which correlated with increased retinoblastoma protein hypophosphorylation. These results show differential redox control of progression from G(1) to S in nonmalignant versus malignant cells and support the hypothesis that loss of a redox control of the cell cycle could contribute to aberrant proliferation seen in cancer cells. PMID- 15890018 TI - p21(Cip1/WAF1/Sdi1) does not affect expression of base excision DNA repair enzymes during chronic oxidative stress. AB - Exposure to chronic oxidative stress during elevated oxygen (hyperoxia) damages DNA and inhibits cell proliferation in G(1) through induction of the cyclin dependent kinase inhibitor p21. Cells that fail to express p21 growth-arrest in S phase. The observation that growth arrest in G(1) is associated with reduced DNA damage and enhanced survival suggests that p21 may affect expression of base excision repair (BER) enzymes used to repair oxidized DNA. This hypothesis was tested in p21 wild-type and p21-deficient mice and human lung adenocarcinoma H1299 cells with tetracycline-on regulated expression of p21. The mRNA levels of Ogg1, Tdg, Udg, Mpg, Nth1, and Mgmt remained constant during 3 days of hyperoxia. The expression of Ogg1, Nth1, and APE protein also remained unchanged. Although hyperoxia increased p21, its absence did not significantly affect expression of these repair enzymes. These findings reveal that hyperoxia induces p21 without significantly altering BER enzyme expression. This suggests that p21 may protect oxidized cells by affecting the activity of BER enzymes and/or through other mechanisms, such as apoptosis. PMID- 15890019 TI - Reactive species-mediated regulation of cell signaling and the cell cycle: the role of MAPK. AB - Cardiovascular disease development is significantly influenced by the effects of reactive species (RS). By virtue of their controlled production, regulation, and reactive nature, RS play important roles in the modulation of cellular signaling, growth, and death in the vasculature. Concentration gradients are important in determining the effects of RS. Low to moderate concentrations of RS act as mediators in signaling cascades and gene regulation, whereas high levels of RS cause cellular damage and death. Because a dual redox regulation state seems to exist in several signaling cascades, e.g., RS often induce upstream initiating events, whereas downstream events are reliant on reductive processes, alterations in cellular redox states influence the activation/inactivation of signaling events and transcription factors. In this review, the relationships between RS, specific signal transduction pathways, and aspects of cell-cycle control are discussed. PMID- 15890020 TI - Redox regulation of cell-cycle re-entry: cyclin D1 as a primary target for the mitogenic effects of reactive oxygen and nitrogen species. AB - Reactive oxygen and nitrogen species inhibit or promote cell proliferation by modulating the cell signaling pathways that dictate decisions between cell survival, proliferation, and death. In the growth factor-dependent pathways that regulate mitogenesis, numerous positive and negative effectors of signaling are influenced by physiological fluctuations of oxidants, including receptor tyrosine kinases, small GTPases, mitogen-activated protein kinases, protein phosphatases, and transcription factors. The same mitogenic pathways that are sensitive to oxidant levels also directly regulate the expression of cyclin D1, a labile factor required for progression through the G1 phase on the cell cycle. Because the transition from G0 to G1 is the only phase of the cell cycle that is not regulated by cyclin-dependent kinases, but rather by redox-dependent signaling pathways, expression of cyclin D1 represents a primary regulatory node for the dose-dependent effects of oxidants on the induction of cell growth. We suggest that expression of cyclin D1 represents a useful marker for assessing the integration of proliferative and growth inhibitory effects of oxidants on the redox-dependent signaling events that control reentry into the cell cycle. PMID- 15890021 TI - FOXO transcription factors in cell-cycle regulation and the response to oxidative stress. AB - Mammalian forkhead members of the class O (FOXO) transcription factors, including FOXO1, FOXO3a, and FOXO4, are implicated in the regulation of a variety of cellular processes, including the cell cycle, apoptosis, DNA repair, stress resistance, and metabolism. FOXO proteins are negatively regulated by the phosphatidylinositol 3-kinase-Akt signaling pathway, which is activated by growth factors and cytokines. Recent studies indicate that the activities of FOXO proteins are also regulated by oxidative stress, which induces their phosphorylation, translocation to the nucleus, and acetylation-deacetylation. Similar to the tumor suppressor p53, FOXO is activated by stress and induces the expression of genes that contribute to cell-cycle arrest, suggesting that it also functions as a tumor suppressor. PMID- 15890022 TI - Redox regulation of the Cdc25 phosphatases. AB - The Cdc25 phosphatases are essential for cell-cycle control in eukaryotes under normal conditions and in response to DNA damage via checkpoint controls. Recent evidence indicates direct control of the Cdc25s, and therefore the cell cycle, in response to changes in cellular redox status. These redox changes may originate intracellularly from mitochondrial leakage or in response to specific external triggers leading to production of reactive oxygen species (ROS). This review shows that the known chemistry and biology of the Cdc25s favor a direct role for these phosphatases in temporarily blocking cell-cycle progression until favorable reducing conditions are restored. First, the Cdc25s contain a highly reactive cysteine at the active site that can react directly with ROS, leading to enzyme inactivation. Second, the ROS-inactivated form of Cdc25 is expected to prevent cell-cycle progression based on precedent from cellular responses to DNA damage. Third, ROS-mediated oxidation of the Cdc25s leads to an intramolecular disulfide that is readily reversible by the cellular reductant thioredoxin. Finally, in vivo data supporting a direct role for the Cdc25s in redox regulation are considered. PMID- 15890024 TI - Hydrogen sulfide as a biological mediator. PMID- 15890023 TI - Peroxiredoxins, oxidative stress, and cell proliferation. AB - Peroxiredoxins (Prxs) are a family of multifunctional antioxidant thioredoxin dependent peroxidases that have been identified in a large variety of organisms. The major functions of Prxs comprise cellular protection against oxidative stress, modulation of intracellular signaling cascades that apply hydrogen peroxide as a second messenger molecule, and regulation of cell proliferation. In the present review, we discuss pertinent findings on the protein structure, the cell- and tissue-specific distribution, as well as the subcellular localization of Prxs. A particular emphasis is put on Prx I, which is the most abundant and ubiquitously distributed member of the mammalian Prxs. Major transcriptional and posttranslational regulatory mechanisms and signaling pathways that control Prx gene expression and activity are summarized. The interaction of Prx I with the oncogene products c-Abl and c-Myc and the regulatory role of Prx I for cell proliferation and apoptosis are highlighted. Recent findings on phenotypical alterations of mouse models with targeted disruptions of Prx genes are discussed, confirming the physiological functions of Prxs for antioxidant cell and tissue protection along with an important role as tumor suppressors. PMID- 15890025 TI - Cadmium exposure alters metabolomics of sulfur-containing amino acids in rat testes. AB - This study aimed to examine distribution of cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CSE), the hydrogen sulfide (H(2)S)-generating enzymes, and metabolomic alterations in sulfur-containing amino acids in rat testes exposed to stressors. Immunohistochemistry revealed distinct distribution of the two enzymes: CBS occurred mainly in Leydig cells and was also detectable in Sertoli cells and germ cells, whereas CSE was evident in Sertoli cells and immature germ cells involving spermatogonia. The amounts of CSE and CBS in testes did not alter in response to administration of cadmium chloride, an antispermatogenic stressor leading to apoptosis. Metabolome analyses assisted by liquid chromatography equipped with mass spectrometry revealed marked alterations in sulfur-containing amino acid metabolism: amounts of methionine and cysteine were significantly elevated concurrently with a decrease in the ratio between S adenosylhomocysteine and Sadenosylmethionine, suggesting expansion of the remethylation cycle and acceleration of methyl donation. Despite a marked increase in cysteine, amounts of H(2)S were unchanged, leading to a remarkable decline of the H(2)S/cysteine ratio in the cadmium-treated rats. Under such circumstances, oxidized glutathione (GSSG) was significantly reduced, whereas reduced glutathione (GSH) was well maintained, and the GSH/GSSG ratio was consequently elevated. These results collectively showed that cadmium induces metabolomic remodeling of sulfur-containing amino acids even when the protein expression of CBS or CSE is not evident. Although detailed mechanisms for such a remodeling event remain unknown, our study suggests that metabolomic analyses serve as a powerful tool to pinpoint a critical enzymatic reaction that regulates metabolic systems as a whole. PMID- 15890026 TI - Hydrogen sulfide as an endogenous modulator of biliary bicarbonate excretion in the rat liver. AB - Cystathionine gamma-lyase (CSE) is an enzyme catalyzing cystathionine and cysteine to yield cysteine and hydrogen sulfide (H(2)S), respectively. This study aimed to examine if H(2)S generated from the enzyme could serve as an endogenous regulator of hepatobiliary function. Gas chromatographic analyses indicated that, among rat organs herein examined, liver constituted one of the greatest components of H(2)S generation in the body, at 100 mumol/g of tissue, comparable to that in kidney and 1.5-fold greater than that in brain, where roles of the gas in the regulation of neurotransmission were reported previously. At least half of the gas amount in the liver appeared to be derived from CSE, because blockade of the enzyme by propargylglycine suppressed it by 50%. Immunohistochemistry revealed that CSE occurs not only in hepatocytes, but also in bile duct. In livers in vivo, as well as in those perfused ex vivo, treatment with the CSE inhibitor induced choleresis by stimulating the basal excretion of bicarbonate in bile samples. Transportal supplementation of NaHS at 30 mumol/L, but not that of N-acetylcysteine as a cysteine donor, abolished these changes elicited by the CSE inhibitor in the perfused liver. The changes elicited by the CSE blockade did not coincide with alterations in hepatic vascular resistance, showing little involvement of vasodilatory effects of the gas in these events, if any. These results first provided evidence that H(2)S generated through CSE modulates biliary bicarbonate excretion and is thus a determinant of bile salt-independent bile formation in the rat liver. PMID- 15890027 TI - Physiological roles of hydrogen sulfide: synaptic modulation, neuroprotection, and smooth muscle relaxation. AB - Nearly 300 years have passed since the first description of the toxicity of hydrogen sulfide (H(2)S) in 1713. Although many studies have been devoted to its toxicity, very little attention has been paid to understanding its normal physiological function. Relatively high concentrations of endogenous H(2)S, however, have recently been discovered in animal tissues, and its possible function as a biological messenger has been proposed. H(2)S enhances the activity of N-methyl-D-aspartate receptors and facilitates the induction of hippocampal longterm potentiation, a synaptic model for memory. H(2)S also increases intracellular concentrations of Ca(2+) in glia and induces Ca(2+) waves, which mediate glial signal transmission. Based on accumulating evidence for the reciprocal interactions between glia and neurons, it has been suggested that glia modulate synaptic transmission. Therefore, H(2)S may regulate synaptic activity by modulating the activity of both neurons and glia. In addition to a role in the signal transduction, H(2)S protects neurons from oxidative stress and in smooth muscle it may function as a relaxant. H(2)S, the toxic gas, may therefore be used as a multifunctional signaling mechanism under normal physiological conditions. PMID- 15890028 TI - Vascular actions of hydrogen sulfide in nonmammalian vertebrates. AB - Hydrogen sulfide (H(2)S) vasoactivity has been observed in isolated vessels from all vertebrate classes, and its effects, which include constriction, dilation, and multiphasic responses, are both species- and vessel-specific. H(2)S is synthesized by mammalian and fish vessels, and because plasma H(2)S titers are also vasoactive in vitro, it is likely that H(2)S is a tonic effector of cardiovascular homeostasis in many vertebrates. Mechanisms of H(2)S vasoactivity in nonmammalian vertebrates have been limited to the trout where the triphasic relaxation-contraction-relaxation includes endothelium-dependent and -independent components, ATP-dependent K(+) channels, and extracellular and intracellular Ca(2+), all independent of cyclic GMP production. The observation that at least some H(2)S constrictory activity has been observed in all vertebrates except sharks suggests that H(2)S may have been an ancestral pressor gasotransmitter. However, the ability of H(2)S to serve as either (or both) an endothelium independent constrictor or dilator, which is relatively unique among vasoregulatory molecules, is a feature that seems to have been exploited, for unknown reasons, by nearly all vertebrates. Aquatic vertebrates appear particularly vulnerable to H(2)S because of their intrinsically low blood pressure and the potential for increased H(2)S exposure from the environment. PMID- 15890029 TI - The role of cystathionine beta-synthase in homocysteine metabolism. AB - Cystathionine beta-synthase (CBS) is the first enzyme in the transsulfuration pathway, catalyzing the conversion of serine and homocysteine to cystathionine and water. The enzyme contains three functional domains. The middle domain contains the catalytic core, which is responsible for the pyridoxal phosphate catalyzed reaction. The C-terminal domain contains a negative regulatory region that is responsible for allosteric activation of the enzyme by S adenosylmethionine. The N-terminal domain contains heme, and this domain regulates the enzyme in response to redox conditions. Besides its canonical reaction, CBS can catalyze alternative reactions that produce hydrogen sulfide, a novel neuromodulator in the brain. Mutations in human CBS result in homocystinuria, an autosomal recessive disorder characterized by defects in a variety of different organ systems. The most common CBS allele is 833T>C (I278T), which is associated with pyridoxine-responsive homocystinuria. A complementation system in S. cerevisiae has been developed for analysis of human CBS mutations. Using this system, it has been discovered that deletion of the C-terminal domain of CBS can suppress the functional defects of many patient-derived mutations. This finding suggests it may be possible to develop drugs that interact with the C-terminal domain of CBS to treat elevated homocysteine in humans. PMID- 15890030 TI - Thioredoxin and its related molecules: update 2005. AB - Studies on thioredoxin (Trx) and its related molecules have expanded dramatically recently. Proteins that share the similar active-site sequence, -Cys-Xxx-Yyy-Cys , are called the Trx family, and the number of Trx family members is increasing. Trx reductase, which reduces oxidized Trx in cooperation with NADPH, has three isoforms, and peroxiredoxin, which is Trx-dependent peroxidase, has six isoforms. In addition to a role as an antioxidant, Trx and its related molecules play crucial roles in the redox regulation of signal transduction. The classical cytosolic Trx1 and truncated Trx80 are released from cells. Plasma/serum levels of Trx1 are good markers for oxidative stress. Exogenous Trx1 shows cytoprotective and antiinflammatory effects and has a good potential for clinical application. This is an update review on Trx and its related molecules. PMID- 15890031 TI - Redox control of growth factor signaling: recent advances in cardiovascular medicine. AB - Growth factors play vital roles in the regulation of various biologic processes, including those in cardiovascular and respiratory systems. Accumulating evidence suggests that reactive oxygen species mediate growth factor signal transduction. The discovery of reactive oxygen species production by angiotensin II in vascular smooth muscle cells via the activation of NAD(P)H oxidase promoted studies of redox control of growth factor signaling. In the past few years, there have been further advances in this field. In addition to established roles of reactive oxygen species in vascular smooth muscle growth, these species have been demonstrated to serve as second messengers for cardiac hypertrophy induced by angiotensin II. NAD(P)H oxidase also produces reactive oxygen species in response to endothelin-1 in vascular smooth muscle and cardiac muscle cells. These results suggest that inhibiting NAD(P)H oxidase might be a useful therapeutic strategy. In fact, adenovirus-mediated gene transfer appears to be an effective approach to prevent vascular hypertrophy in rodent models. Growth factors also induce survival signaling in cardiac and smooth muscle cells, and redox control may play a role in such events. It is likely that studies reporting the mechanisms of redox control of growth factor signaling will rapidly emerge in the next several years, and understanding of such regulation should help in the development of therapeutic strategies against heart and lung diseases. PMID- 15890032 TI - Oxidative protein folding: an update. PMID- 15890034 TI - Is anything changing for the better? PMID- 15890035 TI - The road to formal recognition: the end is in sight. PMID- 15890036 TI - Palliation of brain stem metastasis in patients with lung cancer by radiotherapy: how safe? PMID- 15890038 TI - Palliative care needs of cancer patients in U.S. nursing homes. AB - BACKGROUND: Increasingly, nursing homes are the place of care for older Americans with cancer. Yet, few studies has characterized the quality of care for this growing population. OBJECTIVE: Characterize the scope and quality of cancer care in U.S. nursing homes. DESIGN: Secondary analysis of the national repository of the Minimum Data Set (MDS) SETTING AND SUBJECTS: Nursing home residents noted to have cancer diagnosis on the MDS. RESULTS: Of the 190,769 New Hampshire residents (8.8%) with a cancer diagnosis, 1 in 4 had weight loss (23.4%), received intravenous medications (27.7%), or used oxygen (25.4%). Overall, 45.3% had a do not-resuscitate (DNR) order, with state variations ranging from 17.8% (New Jersey) to 70.5% (Wisconsin). More than 1 in 10 (12.0%) were defined as terminally ill, although only 29.3% of these received hospice services. Among patients with pain, half of those who survived to a second assessment had persistent, severe pain (51.3%), which also varied by state, ranging from 43.3% (Iowa) to 65.8% (Nevada). Active treatment was rare; less than 5% received chemotherapy or radiotherapy. However, 15.5% had parenteral and/or tube feedings for nutrition. Approximately, 1 in 10 New Hampshire residents had advanced cancer. CONCLUSION: Our findings suggest important opportunities to improve the quality of cancer care for older adults. PMID- 15890039 TI - Disclosure of prognosis to terminally ill patients: attitudes and practices among French physicians. AB - BACKGROUND: Disclosure of the prognosis to terminally ill patients is a strong prerequisite for integrating the physical, psychological, spiritual, and social aspects of end-of-life care. OBJECTIVE: This study aimed to assess French physicians' general attitudes and effective practices (with patients followed up to death recently) toward such disclosure. DESIGN/SUBJECTS: We used data from a cross-sectional survey conducted among a national sample of 917 French general practitioners, oncologists, and neurologists. RESULTS: A majority of respondents opted for prognosis disclosure only at patients' request, very few opted for systematic disclosure without patient's request, and a significant minority opted for systematic concealment. Concerning deceased patients described by respondents, 44.5% of competent patients were informed of prognosis by their physician, 27.3% were informed by someone else, 9.0% refused to be informed, and 19.2% were simply not informed. Concealment was more frequent for older patients, and physicians involved in a nongovernmental organization (NGO) for patients' support were more likely to disclose the prognosis, even without patient's request. CONCLUSIONS: Disclosure of the prognosis to terminally ill patients is still far from systematic in France. Further research is needed to better understand the motivations of French physicians' disclosure practices, which are probably culturally shaped. PMID- 15890040 TI - Opportunities and challenges to improving end-of-life care for seriously ill elderly patients: a qualitative study of generalist physicians. AB - BACKGROUND: End-of-life care quality deficiencies have in part been linked to inadequate physician skill in the technical and communication domains of palliative care. Yet few studies have examined physicians' perspectives regarding their experiences caring for patients approaching the end of life. OBJECTIVE: To understand generalist physicians' perspectives regarding their care of seriously ill elderly patients in order to identify challenges to improving end-of-life communication and decision-making. DESIGN: Qualitative study using in-depth ethnographic interviews. SETTING: Puget Sound region of Washington State. PARTICIPANTS: Thirteen community-based generalist physicians who routinely care for elderly patients and represent a range of practice styles and experiences. RESULTS: The physicians described a "revolving door syndrome" in which elderly patients are repeatedly hospitalized with chronic illness exacerbations. Three themes influenced physicians' interactions with "revolving door" patients: (1) physicians' use of decision-making heuristics, characterized as "internal gauges," to promote care consistent with their own values; (2) families' "unreasonable expectations" that patients would return to their previous health after treatment of an acute illness; and (3) families' reluctance to accept end of-life decision-making responsibility. CONCLUSIONS: Our findings suggest that physician values and physician-family interactions impact decision-making for chronically ill elderly patients. The influence of physicians' internal gauges on end-of-life care can facilitate or hinder use of palliative care as well as decision-making consistent with patients' preferences. Disparate physician and family expectations regarding their division of decision-making responsibility and patients' care outcomes may also affect decision-making. The use of communication strategies that promote alignment of these expectations may improve decision-making quality for incapacitated elderly patients. PMID- 15890041 TI - Improving end-of-life care in nursing facilities: the Community State Partnership To Improve End-of-Life Care--California. AB - BACKGROUND: The California Coalition for Compassionate Care formed in 1998 when activists and organizational leaders in several arenas sought to link their efforts for synergistic impact on end-of-life care and to obtain funding to sustain their forward momentum and collaboration. The Coalition focused on public engagement, professional education, and reforms in skilled nursing facilities. With skilled nursing facilities, the Coalition's work built on the efforts of the ECHO (Extreme Care, Human Options) Long Term Care Task Force, which served as a precursor to the Coalition. OBJECTIVE: The Coalition's objective was to assist committed facilities in devising processes of care that would operationalize basic end-of-life care principles in a manner specific to their particular facility. DESIGN: The Coalition recruited three-member leadership teams from nursing facilities throughout California to attend a 2-day training program, write an action plan, and receive 6 months of modest follow-up support. To assess its success, the group used posttraining evaluations, a follow-up evaluation, a focus group, and informal feedback over several years to assess the dynamics, achievements, and challenges of their efforts. RESULTS: In 2000-2002 the training reached 298 people representing 109 nursing facilities and each district office of the nursing facility surveyors. Response to the training was enthusiastic. Self-reported improvements in 27 care practices were best in the areas of pain assessment and management. Completion of nursing facilities' self-identified action plans varied widely. Participants generally perceived the commitment to improving end-of-life care as a vehicle for improving the overall care and quality management in nursing facilities. CONCLUSIONS: The specific challenges of organizational change in nursing facilities require sustained, focused leadership and hands-on guidance to overcome the inevitable barriers and setbacks. The Coalition's experience confirms that coalitions depend upon personal commitments and relationships, a focus on practical products, and a consistent infrastructure. PMID- 15890042 TI - Kokua Mau: a statewide effort to improve end-of-life care. AB - BACKGROUND: Many Americans die in pain, without hospice, and without regard to advance directives, suggesting a need to improve end-of-life (EOL) awareness and services. OBJECTIVE: This paper describes Kokua Mau, a community-state partnership to improve EOL in Hawaii funded by The Robert Wood Johnson Foundation (RWJF). Coalition activities were guided by innovation-diffusion theory, targeting "innovators" and "change agents" within communities and organizations willing to learn about and facilitate improvements to EOL care. DESIGN: Evaluation of a community-wide intervention to improve EOL care. SETTING/SUBJECTS: Honolulu, Hawaii. MEASUREMENTS: We tracked dissemination of campaign messages by counting numbers of coalition members (including innovators and change agents to carry on the work), individuals reached through awareness and educational offerings, and new EOL projects initiated during and after the initial 3-year RWJF funding. To measure change, we counted the number of legislative policies that were modified by the coalition as well as indicators of hospice utilization, advance directive (AD) completion, support for physician assisted death, and place of death. RESULTS: In the first 3 years of the project: coalition membership grew to 350 members; EOL care curricula were developed and offered to various target audiences; 17,000 individuals attended educational events; policy changes were facilitated; decreases were seen in proportions of residents supporting physician-assisted suicide; and increases were seen in advance directive completion rates and hospice utilization. Most importantly, after the grant period, coalition members went on to develop and implement new programs to improve care to the dying. CONCLUSIONS: Although it will take several years to effect comprehensive and sustained changes in the way death is perceived and the dying process is facilitated, findings suggest that programs based on innovation-diffusion theory can increase EOL awareness and help develop the change agents and role models needed to affect community-wide change over the long term. PMID- 15890043 TI - The development and outcomes of a statewide network of hospital-based palliative care teams. AB - BACKGROUND: Health care systems need to be developed that meet the palliative care needs of patients and their families. OBJECTIVE: The West Virginia Initiative to Improve End-of-Life Care charged its Palliative Care Delivery System Task Force with developing hospital-based palliative care teams in West Virginia. SETTING/SUBJECTS: A descriptive study of a state-community partnership to improve palliative care. MEASUREMENTS: Numbers of member hospitals, number and nature of palliative care consultations, number of patients referred to hospice programs. RESULTS: What began as a palliative care network of five hospitals has grown into a network of 16. Network members advise one another on financing, staffing, and obtaining administrative buy-in, and the network leaders provide educational programs to prepare physicians and nurses to serve as consultants on hospital-based palliative care teams. There was a 300% increase in the number of palliative care consultations between 2000 and 2003 in 49 of West Virginia's 55 counties. Analysis of the submitted data collections forms has allowed the network to promote improvement in palliative care consultations in member hospitals and to initiate interventions on a statewide basis to improve decision making with the appropriate legal agent, treatment of pain, and referral of patients for hospice care. CONCLUSION: Hospital participation in the Palliative Care Delivery System Task Force led to a sustained membership organization, the West Virginia Palliative Care Network that promotes hospital-based palliative care. PMID- 15890044 TI - Supportive-affective group experience for persons with life-threatening illness: reducing spiritual, psychological, and death-related distress in dying patients. AB - BACKGROUND: Attention to psycho-socio-spiritual needs is considered critical by patients with life-threatening illnesses and their caregivers. Palliative care interventions that address these needs--particularly spirituality--are lacking. OBJECTIVE: To evaluate the effects of an innovative program to address psycho socio-spiritual needs in patients with life-threatening illnesses. DESIGN: A group intervention entitled Life-Threatening Illness Supportive-Affective Group Experience (LTI-SAGE) was developed for reducing patient spiritual, emotional, and death-related distress. SETTING/SUBJECTS: African American and Caucasian patients (n = 69) from two hospitals in St. Louis, Missouri, with life threatening medical conditions (cancer; human immunodeficiency virus/acquired immune deficiency syndrome [HIV/AIDS]; geriatric frailty; liver, kidney, pulmonary, or cardiovascular disease) were randomly assigned to intervention or control groups. Intervention patients participated in a maximum of 12 LTI-SAGE groups over a 12-month period. Control patients received standard care. MEASUREMENTS: Outcome measures were depression symptoms, anxiety, spiritual well being, and death-related emotional distress. RESULTS: After attrition, 51 (73.9%) patients completed the trial. At the end of the trial, after factoring in compliance, intervention patients had significantly fewer depression symptoms and death-related feelings of meaninglessness and significantly better spiritual well being than did control patients. CONCLUSIONS: The use of the LTI-SAGE model for enhancing the end-of-life illness experience is promising. PMID- 15890045 TI - Pilot of a hospice-based elective to learn comfort with dying patients in undergraduate medical education. AB - BACKGROUND: A majority of medical students feel uncomfortable with dying patients. OBJECTIVE: We designed a pilot program to train medical students to be hospice volunteers to determine (1) the value of contact with dying patients in changing medical students' comfort level and (2) the mechanics of starting such a course in cooperation with a nonacademic agency. DESIGN: Students were required to undergo hospice training provided by the local hospice. Students were eligible for Hospice volunteer placement after the training was completed. This provided students with an opportunity to interact with dying patients in a nonclinical capacity. Quantitative data and qualitative data were collected about the students' experiences with patients as well as about the effectiveness of the rotation. SETTING/SUBJECTS: First- through fourth-year medical students were recruited to participate in this semester long project via a medical school-wide e-mail. MEASUREMENTS: (1) A self-rating questionnaire on emotions and attitudes about death and dying at three points in time: before training, after training, and after placement; (2) A Likert-type questionnaire on barriers to participation and the usefulness of the components of an end-of-life course; (3) The "Attitudes About End-of-Life Scale"; (4) An in-depth structured interview with students after completion of the course. RESULTS: Fourteen students participated in the evaluation phase of the program and reported increased comfort in interacting with dying patients after participating in the program. Students had less anxiety and fear of being around a dying person after placement as compared to before training. In-depth student interviews provided valuable qualitative data on the impact of the pilot, and insight into the strengths and areas for improvement in this type of elective. CONCLUSIONS: A Hospice based elective can be an effective model for facilitating learning about how to approach the patient with a terminal illness. PMID- 15890046 TI - An undergraduate course on palliative medicine and end-of-life issues. AB - BACKGROUND: The concept and development of this course were the joint efforts of the directors of the Susquehanna University Office of the Chaplain and the Geisinger Health System Palliative and Supportive Medicine Program (PMP). Both individuals perceived a need for increased awareness of students for issues related to advancing age and terminal illness. OBJECTIVES: Our objectives in formulating and presenting this course were to demystify aging, illness and death; increase awareness and tolerance for cultural and religious differences; enhance empathy and teach compassion; provide communication skills that could be used in many difficult situations; and provide a caring and mutually beneficial relationship for the elderly and students in our community. DESIGN: The one semester course consisted of 28 one-hour forty-five minute classes and a separate service-learning project. One half of the lectures, presented by the Susquehanna University chaplain (M.R.), predominantly involved religious and spiritual issues. The other 14 were organized by the PMP and were presented primarily by health care professionals. A required service-learning project was the creation of a "personal legacy." SETTINGS/SUBJECTS: The PMP group comprised 7 physicians, 3 nurses, 1 chaplain, and 2 individuals participating in volunteer community activities for the elderly and infirm. All volunteered their time for this endeavor. Each had autonomy concerning his class content and format. Eighteen students (2 seniors, 6 juniors, 7 sophomores and 3 freshmen) registered for and completed the course. MEASUREMENTS: In the final week, students were asked to evaluate the course with predominantly open-ended questions. They were informed that they would not be graded on their responses, and complete candor was encouraged. A less detailed evaluation of the overall experience by the course instructors was also solicited. These various comments are reviewed and discussed. RESULTS AND CONCLUSIONS: Overall, student and faculty comments about the course were quite positive. Every student believed that it was a very worthwhile course, but 4 of 18 would not sign up for it again and would not recommend it to a friend. We used the comments from the student and faculty participants to change the curriculum and presentation style for future courses. PMID- 15890047 TI - Assessing competence of residents to discuss end-of-life issues. AB - BACKGROUND: Residents are often responsible for eliciting patients' treatment preferences at the end of life (EOL), yet we have a limited understanding of their competence in this task. OBJECTIVE: To assess the competence of medical residents to discuss advance directives (AD) with patients using two measures: self-assessment (perceived competence) and self-reported behaviors (behavioral competence). To examine the relationship between educational experiences and these two measures of competence. DESIGN: Cross-sectional self-report questionnaire. SUBJECTS: Internal medicine residents from two university- and one community-based program. RESULTS: The 282 respondents (84% response) had an average of 6.2 EOL discussions per month. Few residents reported having received useful feedback from a resident (8%) or an attending (7%) about their ability to discuss ADs. Even fewer reported that work rounds (4%) or attending rounds (5%) were frequently forums for learning about EOL care. Mean perceived competence was 3.8 (range, 1-5). In multivariable analyses, greater perceived competence was significantly associated with higher postgraduate year (p < 0.001), having residents demonstrate exemplary AD discussions (p < 0.001), and less formal education (p < 0.01). Behavioral competence was significantly associated with reporting that work rounds were useful for learning about EOL care (p = 0.002), less formal education (p = 0.02) and a greater number of EOL discussions per ward month (p = 0.009). The correlation between perceived and behavioral competence (r = 0.25, p = 0.001) was modest but statistically significant. CONCLUSIONS: Many residents view themselves as competent to discuss ADs with patients but fail to engage in recommended behaviors for such discussions. Increasing experiential learning may be the most promising means of enhancing residents' abilities to discuss EOL issues with patients. PMID- 15890048 TI - Dying, death, and medical education: student voices. AB - BACKGROUND: Medical schools require time for end-of-life topic. However, there is very little medical literature that directly addresses how medical students and residents are to behave, manage emotion, and confront their own grieving process when patients die. OBJECTIVE: The purpose of this study was to understand how preclinical medical students describe feelings toward the death of a hypothetical patient in order to affect curricular change at our institution. DESIGN: Qualitative methods using narrative analysis of student papers to identify patterns, core constructs, and themes related to student's projected feelings on patient death. SETTING/SUBJECTS: Federal medical school with volunteer medical students from the class of 2005. RESULTS: Two thirds of the students (108/162) volunteered to participate. Five significant themes emerged including: (1) affective responses (guilt, fear, blame, impotence), (2) personal experience with death, (3) survivorship and professionalism, (4) the meaning of death, and (5) the affects of religion and spirituality. Many feared facing families and responding to grief. An active belief in an afterlife was mentioned as a coping strategy by 40% of the students. CONCLUSIONS: End-of-life curriculum is more than teaching about the clinical care of the patient and support of family. These medical students overwhelmingly identified the need for coping strategies when confronting the dying patient. Teaching students these coping strategies should be an integral part of an end-of-life curriculum. Writing exercises cannot only help students recognize and reflect upon their emotions and feelings, but also allow educators a window into curricular elements that need to be added to death and dying education. PMID- 15890049 TI - Healing the healer: poetry in palliative care. AB - BACKGROUND: Poetry plays an age-old role in the art of healing. Although medicine today seems distant from the world of poetic expression, there are surprising commonalities between the two. OBJECTIVES: In this essay we reflect on three aspects of healing that are fostered by poetry. OBSERVATIONS: Practicing medicine with too many facts and not enough poetry leads to dissatisfaction, disappointment, and impaired healing, especially in the care of the terminally ill. Likewise, poetry deficiency cuts off an important avenue for physician self awareness and reflectivity. Alternatively, three aspects of healing are fostered by poetry: the power of the word to heal (and also harm); the skill of "negative capability" that enhances physician effectiveness; and empathic connection, or compassionate presence, a relationship that heals without words. CONCLUSION: Reading and writing poetry can help physicians, especially those who care for dying patients, become more reflective, creative, and compassionate practitioners. PMID- 15890051 TI - Spiritual, religious, and existential aspects of palliative care. PMID- 15890052 TI - Series introduction: The profession of social work in end-of-life and palliative care. PMID- 15890053 TI - A national agenda for social work research in palliative and end-of-life care. AB - BACKGROUND: Social work practitioners have the potential to make meaningful contributions to improving palliative and end-of-life care because of their work in varied and divergent practice settings across the lifespan, their role in addressing mental health needs, grief and psychosocial aspects of well-being, and their commitment to promoting culturally competent, effective, and humane care, particularly for the most vulnerable and oppressed members of society. The federal government and several national and professional institutes have called for steps to increase the participation of social work researchers as well as to improve the quality, quantity, and dissemination of social work research. OBJECTIVE: This paper proposes a national agenda for social work research in palliative and end-of-life care. DESIGN: The agenda was developed by a multimethod effort that included an extensive analysis of the literature, discussions with a purposive national sample of social work leaders in palliative or end-of-life care, and consideration of established national research priorities. RESULTS: Eleven domains of palliative and end-of-life care and their related research objectives were identified. Their relevance to the mission and value base of the profession and to established national research priorities is highlighted. CONCLUSIONS: This research agenda should serve to stimulate social work research initiatives to improve palliative and end-of-life care, and ultimately inform direct practice, policy and professional education. PMID- 15890054 TI - An approach to develop effective health care decision making for women in prison. AB - BACKGROUND: Decision making about medical treatment, advance care planning and end-of-life care often is limited or influenced by a patient's capacity to read or comprehend crucial information. Ineffective communication between patient and physician, and the complex nature of serious illness and dying also affect these decisions. America's incarcerated have virtually no autonomy in decision making, especially with regard to medical care and treatment. For the nearly 100,000 incarcerated women in the United States, medical issues differ significantly from those of male prisoners. Many women enter prison with chronic illnesses or are diagnosed with such illnesses while in prison. In addition, America's prison population reflects our country's unsolved literacy problems with almost two thirds of aging inmates lacking basic literacy skills. Maintaining a balance between the Eighth Amendment rights of prisoners and their status as wards of the state is a concern for inmates and for those responsible for their care. OBJECTIVE: The purpose of this study was to identify informational barriers to people making medical care and treatment decisions, particularly those with low literacy. Findings were used to in the development of a tool to assist patients to initiate discussion and become active participants in their own care. DESIGN: Female inmates of diverse ethnicity and literacy levels were recruited through self-selection from the Central California Women's Facility near Sacramento to participate in focus group discussions. The focus groups were guided by a set of research questions and were facilitated by the first author to identify informational barriers to medical care needs. SETTING: The female prison setting was identified as an appropriate setting for the development of a new approach to stimulate effective decision making by introducing basic information on medical care and treatment, advance care planning, and end-of-life care. A total of 113 inmates participated in 16 focus group, each 2 hours in length. RESULTS: Findings were used to develop a handbook to assist low literacy populations in general, and female inmates in particular, to obtain appropriate medical care and treatment and to make advance care decisions regarding the end of life. CONCLUSION: Those who face chronic, potentially life-limiting illness cannot make meaningful decisions regarding medical care and treatment without having a basic foundation of health information. Acquiring knowledge and improving communication skills reduce stress and vulnerability, assuring individuals of some control over decision making. PMID- 15890055 TI - Describing the social environment of elderly persons at the time of death. AB - OBJECTIVES: To describe the social environment of elderly persons at the time of their death and its correlates. DESIGN: Telephone interviews with the next-of-kin of deceased elderly persons. PARTICIPANTS: Potential participants were the next of-kin of all Jewish elderly individuals who died during the course of one year in a major city in the northern part of Israel. The final sample included 139 participants (58% response rate; 70% cooperation rate). MEASUREMENTS: Information was collected regarding the decedents' social environment three days before their death and at the time of death. Eight types of informal sources of support and four sources of formal support were examined. The correlates examined included the decedents' demographic, health, social, and death characteristics. RESULTS: The majority of the decedents died while surrounded by at least one family member. Having a larger social network was associated with a larger number of persons present at the time of death. Being informed about the impending death was associated with a larger number of persons from the formal and informal networks present at the time of death. CONCLUSIONS: The common fear of dying alone and isolated was not confirmed by this study. Increased efforts should be invested in helping professionals to deal with the difficulties inherent to the process of diagnosing an upcoming death. PMID- 15890056 TI - Interventional options for malignant upper gastrointestinal obstruction. PMID- 15890057 TI - Diarrhea in palliative care. PMID- 15890059 TI - Panic, anxiety, and chronic dyspnea. PMID- 15890063 TI - JPM patient information. Introduction to palliative and hospice care. PMID- 15890065 TI - Cloning and characterization of the human glutathione synthetase 5'-flanking region. AB - GSH synthesis occurs through a two-step enzymatic reaction driven by GCL (glutamate-cysteine ligase; made up of catalytic and modifying subunits) and GSS (glutathione synthetase). In humans, oxidative stress regulates GCL expression in an antioxidant response element-dependent manner via Nrf2 [NFE (nuclear factor erythroid)-related factor 2]. In the rat, GSS and GCL are regulated co-ordinately by oxidative stress, and induction of GSS further increases GSH synthetic capacity. Transcriptional regulation of the human GSS has not been examined. To address this, we have cloned and characterized a 2.2 kb 5'-flanking region of the human GSS. The transcriptional start site is located 80 nt upstream of the translation start site. The human GSS promoter efficiently drove luciferase expression in Chang cells. Overexpression of either Nrf1 or Nrf2 induced the GSS promoter activity by 130 and 168% respectively. Two regions homologous to the NFE2 motif are demonstrated to be important for basal expression of human GSS, as mutation of these sites reduced the promoter activity by 66%. Nrf1, Nrf2 and c Jun binding to these NFE2 sites under basal conditions was demonstrated using chromatin immunoprecipitation assays. In summary, two NFE2 sites in the human GSS promoter play important roles in the basal expression of GSS and, similar to the GCL subunits, the human GSS gene expression is also regulated by Nrf2. PMID- 15890066 TI - Mutational analysis of the PITX2 coding region revealed no common cause for transposition of the great arteries (dTGA). AB - BACKGROUND: PITX2 is a bicoid-related homeodomain transcription factor that plays an important role in asymmetric cardiogenesis. Loss of function experiments in mice cause severe heart malformations, including transposition of the great arteries (TGA). TGA accounts for 5-7% of all congenital heart diseases affecting 0.2 per 1000 live births, thereby representing the most frequent cyanotic heart defect diagnosed in the neonatal period. METHODS: To address whether altered PITX2 function could also contribute to the formation of dTGA in humans, we screened 96 patients with dTGA by means of dHPLC and direct sequencing for mutations within the PITX2 gene. RESULTS: Several SNPs could be detected, but no stop or frame shift mutation. In particular, we found seven intronic and UTR variants, two silent mutations and two polymorphisms within the coding region. CONCLUSION: As most sequence variants were also found in controls we conclude that mutations in PITX2 are not a common cause of dTGA. PMID- 15890067 TI - Quality of life, functional outcome, and voice handicap index in partial laryngectomy patients for early glottic cancer. AB - BACKGROUND: In this study, we aim to gather information about the quality of life issues, functional outcomes and voice problems facing early glottic cancer patients treated with the surgical techniques such as laryngofissure cordectomy, fronto-lateral laryngectomy, or cricohyoidopexi. In particular, consistency of life and voice quality issues with the laryngeal tissue excised during surgery is examined. In addition, the effects of arytenoidectomy to the life and voice quality are also studied. METHODS: 29 male patients were enrolled voluntarily in the study. The average age was 53.9 years. Three out of 10 patients with laryngofissure cordectomy also had arytenoidectomy. 11 patients had fronto lateral laryngectomy with Tucker reconstruction, two of which also had arytenoidectomy. There were eight patients with cricohyoidopexi and bilateral functional neck dissection. Three of these patients also had arytenoidectomy. In bilateral functional neck dissection cases, spinal accessory nerve was preserved and level V of the neck was not dissected. None of the patients had neither radiotherapy nor voice therapy. Cordectomy patients never had a temporary tracheotomy or were connected to a feeding tube. Data was collected for 13 months for the cordectomy group, 14 months for fronto-lateral laryngectomy and cricohyoidopexi groups on average post-operatively. Statistical analysis in this study was carried out using the one-way analysis of variance, and the Post-Hoc group comparisons were made after Bonferroni and Scheffe-procedures.In order to determine the effects of arytenoidectomy, a regression analysis is carried out to see if there are statistical differences in answers given to the survey questions among patients who were arytenoidectomized during their surgeries. RESULTS: There was a statistically significant difference between cordectomy and cricohyoidopexi group in answers to the University of Washington- Quality of Life- Revised survey part 1. (p = 0). A statistically significant difference was also established between cordectomy and fronto-lateral laryngectomy groups, as well as between cordectomy and cricohyoidopexi groups in answers to the University of Washington- Quality of Life- Revised survey part 2. (p = 0,036 and p = 0.009, respectively). Cricohyoidopexi group has given the lowest scores and the cordectomy group has given the highest scores in three survey questions representing the quality of life, performances and new voices. These ranges are also consistent with the laryngeal tissue excised during surgery (cricohyoidopexi > fronto-lateral laryngectomy > cordectomy). There was no statistically significant difference between groups in Performance Status Scale for Head and Neck cancer patients instrument. The difference between the Voice Handicap Index and Voice Handicap Index (functional); Voice Handicap Index (physical) and Voice Handicap Index (emotional) scores in three patient groups was not significant either. All of the patients evaluated that their new voices have similar functional, physical and emotional impact on their life. Decanulation and oral feeding times of cricohyoidopexi and fronto-lateral laryngectomy patients are found to be significantly longer than cordectomy patients. Lastly, the removal of arytenoid does not have any significant adverse effects on the quality of life, the functional outcomes, or the quality of voice. CONCLUSION: In the present study, all patients with early glottic cancer, treated with different surgical technics reported fairly good quality of life outcomes, functional results and voice qualities. This study also finds that the removal of arytenoid does not have any adverse effects on the quality of life and voice from the patients' point of view. PMID- 15890068 TI - CAGER: classification analysis of gene expression regulation using multiple information sources. AB - BACKGROUND: Many classification approaches have been applied to analyzing transcriptional regulation of gene expressions. These methods build models that can explain a gene's expression level from the regulatory elements (features) on its promoter sequence. Different types of features, such as experimentally verified binding motifs, motifs discovered by computer programs, or transcription factor binding data measured with Chromatin Immunoprecipitation (ChIP) assays, have been used towards this goal. Each type of features has been shown successful in modeling gene transcriptional regulation under certain conditions. However, no comparison has been made to evaluate the relative merit of these features. Furthermore, most publicly available classification tools were not designed specifically for modeling transcriptional regulation, and do not allow the user to combine different types of features. RESULTS: In this study, we use a specific classification method, decision trees, to model transcriptional regulation in yeast with features based on predefined motifs, automatically identified motifs, ChlP-chip data, or their combinations. We compare the accuracies and stability of these models, and analyze their capabilities in identifying functionally related genes. Furthermore, we design and implement a user-friendly web server called CAGER (Classification Analysis of Gene Expression Regulation) that integrates several software components for automated analysis of transcriptional regulation using decision trees. Finally, we use CAGER to study the transcriptional regulation of Arabidopsis genes in response to abscisic acid, and report some interesting new results. CONCLUSION: Models built with ChlP-chip data suffer from low accuracies when the condition under which gene expressions are measured is significantly different from the condition under which the ChIP experiment is conducted. Models built with automatically identified motifs can sometimes discover new features, but their modeling accuracies may have been over-estimated in previous studies. Furthermore, models built with automatically identified motifs are not stable with respect to noises. A combination of ChlP-chip data and predefined motifs can substantially improve modeling accuracies, and is effective in identifying true regulons. The CAGER web server, which is freely available at http://cic.cs.wustl.edu/CAGER/, allows the user to select combinations of different feature types for building decision trees, and interact with the models graphically. We believe that it will be a useful tool to facilitate the discovery of gene transcriptional regulatory networks. PMID- 15890069 TI - Effects of mycophenolate mofetil on key pattern of coronary restenosis: a cascade of in vitro and ex vivo models. AB - BACKGROUND: Mycophenolate mofetil (MMF), the prodrug of mycophenolic acid (MPA), is a rationally designed immunosuppressive drug. The current study investigates the effect of MMF on key pattern of restenosis in a cascade of in vitro and ex vivo models. METHODS: Part I of the study investigated in northern blot and cytoflow studies the effect of MMF (50, 100, 150, 200, 250, and 300 microg/mL) on TNF-alpha induced expression of intercellular adhesion molecule 1 (ICAM-1) in human coronary endothelial cells (HCAEC) and human coronary medial smooth muscle cells (HCMSMC). Part II of the study applied a human coronary 3D model of leukocyte attack, the 3DLA-model. HCAEC and HCMSMC were cultured on both sides of a polycarbonate filters, mimicking the internal elastic membrane. Leukocyte attack (LA) was carried out by adding human monocytes (MC) on the endothelial side. The effect of MMF (50 microg/mL) on adhesion and chemotaxis (0.5, 1, 2, 3, 4, 6, and 24 h after LA) and the effect on proliferation of co-cultured HCMSMC (24 h after LA) was studied. In part III of the study a porcine coronary organ culture model of restenosis (POC-model) was used. After ex vivo ballooning MMF (50 microg/mL) was added to the cultures for a period of 1, 2, 3, 4, 5, 6, and 7 days. The effect on reactive cell proliferation and neointimal thickening was studied at day 7 and day 28 after ballooning. RESULTS: Expression of ICAM-1 in northern blot and cytoflow studies was neither clearly inhibited nor stimulated after administration of MMF in the clinical relevant concentration of 50 microg/mL. In the 3DLA-model 50 microg/mL of MMF caused a significant antiproliferative effect (p < 0.001) in co-cultured HCMSMC but had no effect on MC-adhesion and MC-chemotaxis. In the ex vivo POC-model neighter reactive cell proliferation at day 7 nor neointimal hyperplasia at day 28 were significantly inhibited by MMF (50 microg/mL). CONCLUSION: Thus, the data demonstrate a significant antiproliferative effect of clinical relevant levels of MMF (50 microg/mL) in the 3DLA-model. The antiproliferative effect was a direct antiproliferative effect that was not triggered via reduced expression of ICAM-1 or via an inhibition of MC-adhesion and chemotaxis. Probably due to technical limitations (as e.g. the missing of perfusion) the antiproliferative effect of MMF (50 microg/mL) could not be reproduced in the coronary organ culture model. A cascade of focused in vitro and ex vivo models may help to gather informations on drug effects before large experimental studies are initiated. PMID- 15890070 TI - Cost-analysis of different management policies for patients with mild hepatitis A virus infection in Kazakhstan. AB - OBJECTIVE: For patients with mild hepatitis A virus (HAV) infection, this study compared estimates of total costs associated with managing cases under a policy of mandatory hospitalization in the Republic of Kazakhstan and estimates of total costs associated with managing cases in outpatient settings. Costs were estimated both from the perspective of the Ministry of Health and from a broader societal perspective. METHODS: Data were collected by using a standardized structured questionnaire. For cases of mild HAV infection, medical records were obtained from 200 patients managed by hospitalization and from 251 patients managed in an outpatient setting. Personal interviews were also conducted to collect information on productivity losses and out-of-pocket expenses. RESULTS: Nationally, we estimated about 21,600 cases of mild HAV infection annually. The mean annual treatment costs in hospital for mild HAV infection was estimated at 3.39 million US dollars (2001 US dollars) (95% confidence interval [CI] = [3.26 million US dollars-3.52 million US dollars]). The total annual mild HAV infection cost to the society, including direct medical and nonmedical costs and productivity losses due to 721,440 lost work days, was estimated at 6.26 million US dollars (95% CI [6.05 million US dollars-6.47 million US dollars]). In sensitivity analyses, the total annual cost of mild HAV infection ranged from 4.37 million US dollars to 24.66 million US dollars. The survey results showed that a relatively minor change in the current policy of mandatory hospitalization could result in an estimated total annual savings of 4.62 million US dollars (2001 US dollars) in Kazakhstan. CONCLUSION: Adoption of an outpatient management policy for cases of mild HAV infection would generate substantial cost savings to the Ministry of Health and society. PMID- 15890071 TI - GPs' perspectives of type 2 diabetes patients' adherence to treatment: A qualitative analysis of barriers and solutions. AB - BACKGROUND: The problem of poor compliance/adherence to prescribed treatments is very complex. Health professionals are rarely being asked how they handle the patient's (poor) therapy compliance/adherence. In this study, we examine explicitly the physicians' expectations of their diabetes patients' compliance/adherence. The objectives of our study were: (1) to elicit problems physicians encounter with type 2 diabetes patients' adherence to treatment recommendations; (2) to search for solutions and (3) to discover escape mechanisms in case of frustration. METHODS: In a descriptive qualitative study, we explored the thoughts and feelings of general practitioners (GPs) on patients' compliance/adherence. Forty interested GPs could be recruited for focus group participation. Five open ended questions were derived on the one hand from a similar qualitative study on compliance/adherence in patients living with type 2 diabetes and on the other hand from the results of a comprehensive review of recent literature on compliance/adherence. A well-trained diabetes nurse guided the GPs through the focus group sessions while an observer was attentive for non verbal communication and interactions between participants. All focus groups were audio taped and transcribed for content analysis. Two researchers independently performed the initial coding. A first draft with results was sent to all participants for agreement on content and comprehensiveness. RESULTS: General practitioners experience problems with the patient's deficient knowledge and the fact they minimize the consequences of having and living with diabetes. It appears that great confidence in modern medical science does not stimulate many changes in life style. Doctors tend to be frustrated because their patients do not achieve the common Evidence Based Medicine (EBM) objectives, i.e. on health behavior and metabolic control. Relevant solutions, derived from qualitative studies, for better compliance/adherence seem to be communication, tailored and shared care. GPs felt that a structured consultation and follow-up in a multidisciplinary team might help to increase compliance/adherence. It was recognized that the GP's efforts do not always meet the patients' health expectations. This initiates GPs' frustration and leads to a paternalistic attitude, which may induce anxiety in the patient. GPs often assume that the best methods to increase compliance/adherence are shocking the patients, putting pressure on them and threatening to refer them to hospital. CONCLUSION: GPs identified a number of problems with compliance/adherence and suggested solutions to improve it. GPs need communication skills to cope with patients' expectations and evidence based goals in a tailored approach to diabetes care. PMID- 15890072 TI - Molecular mapping of periodontal tissues using infrared microspectroscopy. AB - BACKGROUND: Chronic periodontitis is an inflammatory disease of the supporting structures of the teeth. Infrared microspectroscopy has the potential to simultaneously monitor multiple disease markers, including cellular infiltration and collagen catabolism, and hence differentiate diseased and healthy tissues. Therefore, our aim was to establish an infrared microspectroscopy methodology with which to analyze and interpret molecular maps defining pathogenic processes in periodontal tissues. METHODS: Specific key cellular and connective tissue components were identified by infrared microspectroscopy and using a chemical imaging method. RESULTS: Higher densities of DNA, total protein and lipid were revealed in epithelial tissue, compared to the lower percentage of these components in connective tissue. Collagen-specific tissue mapping by infrared microspectroscopy revealed much higher levels of collagen deposition in the connective tissues compared to that in the epithelium, as would be expected. Thus inflammatory events such as cellular infiltration and collagen deposition and catabolism can be identified by infrared microspectroscopy. CONCLUSION: These results suggest that infrared microspectroscopy may represent a simple, reagent free, multi-dimensional tool with which to examine periodontal disease etiology using entirely unprocessed tissue sections. PMID- 15890073 TI - BCG directly induces cell cycle arrest in human transitional carcinoma cell lines as a consequence of integrin cross-linking. AB - BACKGROUND: Current models of the mechanism by which intravesical BCG induces an anti-tumor effect in urothelial carcinoma propose a secondary cellular immune response as principally responsible. Our group has demonstrated that BCG mediated cross-linking of alpha51 integrin receptors present on the tumor surface elicits a complex biologic response involving AP1 and NF-kappaB signaling as well as the transactivation of immediate early genes. This study evaluated the direct biologic effect of cross-linking alpha5beta1 integrin on cell cycle progression and apoptosis in two human urothelial carcinoma cell lines. METHODS: Two independent assays (MTT and Colony forming ability) were employed to measure the effect of alpha5beta1 cross-linking (antibody mediated or BCG) on cellular proliferation. Flow cytometry was employed to measure effect of BCG and alpha5beta1 antibody mediated cross-linking on cell cycle progression. Apoptosis was measured using assays for both DNA laddering and Caspase 3 activation. RESULTS: Results demonstrate that integrin cross-linking by BCG, or antibody mediated crosslinking of alpha5beta1 resulted in a decrease in proliferating cell number. BCG treatment or alpha5beta1 cross-linking increased the percentage of cells in G0/G1, in both 253J and T24 cell lines. Peptide mediated blockade of integrin binding site using RGDS reversed the effect BCG on both proliferation and cell cycle arrest. Apoptosis in response to BCG was not identified by either DNA laddering or Caspase 3 activation. CONCLUSION: These findings show that BCG exerts a direct cytostatic effect on human urothelial carcinoma cell lines. Cell cycle arrest at the G1/S interface is a mechanism by which BCG inhibits cellular proliferation. This effect is duplicated by antibody mediated cross-linking of alpha5beta1 and likely occurs as a consequence of crosslink-initiated signal transduction to cell cycle regulatory genes. PMID- 15890074 TI - The NMDA antagonist memantine affects training induced motor cortex plasticity--a study using transcranial magnetic stimulation. AB - BACKGROUND: Training of a repetitive synchronised movement of two limb muscles leads to short-term plastic changes in the primary motor cortex, which can be assessed by transcranial magnetic stimulation (TMS) mapping. We used this paradigm to study the effect of memantine, a NDMA antagonist, on short-term motor cortex plasticity in 20 healthy human subjects, and we were especially interested in possible differential effects of different treatment regimens. In a randomised double-blinded cross over study design we therefore administered placebo or memantine either as a single dosage or as an ascending dosage over 8 days. Before and after one hour of motor training, which consisted of a repetitive co contraction of the abductor pollicis brevis (APB) and the deltoid muscle, we assessed the motor output map of the APB muscle by TMS under the different conditions. RESULTS: We found a significant medial shift of the APB motor output map after training in the placebo condition, indicating training-induced short term plastic changes in the motor cortex. A single dosage of memantine had no significant effect on this training-induced plasticity, whereas memantine administered in an ascending dosage over 8 days was able to block the cortical effect of the motor training. The memantine serum levels after 8 days were markedly higher than the serum levels after a single dosage of memantine, but there was no individual correlation between the shift of the motor output map and the memantine serum level. Besides, repeated administration of a low memantine dosage also led to an effective blockade of training-induced cortical plasticity in spite of serum levels comparable to those reached after single dose administration, suggesting that the repeated administration was more important for the blocking effect than the memantine serum levels. CONCLUSION: We conclude that the NMDA-antagonist memantine is able to block training-induced motor cortex plasticity when administered over 8 days, but not after administration of a single dose. This differential effect might be mainly due to the prolonged action of memantine at the NMDA receptor. These findings must be considered if clinical studies are designed, which aim at evaluating the potency of memantine to prevent "maladaptive" plasticity, e.g. after limb amputation. PMID- 15890075 TI - The immunogenicity of colorectal cancers with high-degree microsatellite instability. AB - BACKGROUND: High-degree microsatellite instability (MSI-H) is a feature of approximately 15% of sporadic colorectal cancers. Patients with MSI-H cancers have been reported to have a better prognosis than those with non-MSI-H cancers. The MSI-H subset is also characterised by a dense infiltrate of intra-epithelial lymphocytes and the hypothesis that the latter represents an efficacious immune response contributing to improved outcome is very attractive. METHODS: Data for this review were identified by searches of MEDLINE, PubMed, and cross references from relevant articles using the search terms 'microsatellite instability', 'colorectal cancer' and 'immunology', 'immune response' or 'immunogenicity'. RESULTS: A total of 38 articles were identified by the search criteria and a further 95 articles by cross-referencing. The relevance of the articles to be interviewed was established by hand searching. Out of a total of 133 articles identified, 47 articles were rejected due to lack of relevance. A total of 86 articles were included in the review, pertaining to microsatellite instability in colorectal cancer, and immune mechanisms in colorectal cancer. CONCLUSION: It is suggested that this distinct group of colorectal cancers may have inherent immunogenic properties and that further elucidation of these may be invaluable to the development of successful immunotherapy. PMID- 15890076 TI - Relationship between maternal obesity and infant feeding-interactions. AB - BACKGROUND: There are no data regarding the relationship between maternal adiposity and interaction and feeding of infants and possible contribution to childhood obesity. In this study we determined the relationship between maternal body weight and composition and infant feeding patterns and maternal-infant interaction during 24-hour metabolic rate measurements in the Enhanced Metabolic Testing Activity Chamber (EMTAC). METHODS: The amount of time four obese (BMI = 33.5 +/- 5.3 kg/m2) and three normal weight (BMI = 23.1 +/- 0.6 kg/m2) biological mothers, spent feeding and interacting with their infants, along with what they ingested, was recorded during 24-hour metabolic rate measurements in the EMTAC. The seven infants were 4.9 +/- 0.7 months, 69 +/- 3 cm, 7.5 +/- 0.8 kg, 26 +/- 3 % fat and 29 +/- 25 percentile for weight for length. Energy and macronutrient intake (kcal/kg) were assessed. Maternal body composition was determined by air displacement plethysmorgraphy and that of the infants by skin-fold thicknesses. Pearson correlations and independent t-tests were utilized for statistical analysis (p < 0.05). RESULTS: Infants born to obese biological mothers consumed more energy (87.6 +/- 18.9 vs. 68.1 +/- 17.3) and energy as carbohydrate (25 +/- 6 vs.16 +/- 3; p < 0.05) than their normal weight counterparts. Most of the increased intake was due to complementary feedings. Twenty-four hour infant energy intake increased with both greater maternal body weight (r = 0.73;p < 0.06) and percent body fat. Furthermore, obese biological mothers spent less total time interacting (570 +/- 13 vs. 381 +/- 30 minutes) and feeding (298 +/- 32 vs.176 +/- 22 minutes) (p < 0.05) their infants than their normal weight counterparts. Twenty-four hour interaction time negatively correlated with both maternal body weight (r = -0.98; p < 0.01) and percent body fat (r = -0.92; p < 0.01). Moreover, infants of obese mothers slept more (783 +/- 38 vs. 682 +/- 32 minutes; p < 0.05) than their normal weight counterparts. However, there were no differences in total 24-hour energy expenditure, resting and sleeping metabolic rates (kcal/kg) for infants born to obese and normal weight biological mothers. CONCLUSION: Greater maternal body weight and percent body fat were associated with greater infant energy intakes. These infants were fed less frequently and consumed more carbohydrates in a shorter period of time as compared to infants from normal weight biological mothers. These variations in feeding patterns may predispose certain infants to obesity. PMID- 15890077 TI - Culture of skeletal myoblasts from human donors aged over 40 years: dynamics of cell growth and expression of differentiation markers. AB - BACKGROUND: Local myogenesis, neoangiogenesis and homing of progenitor cells from the bone marrow appear to contribute to repair of the infarcted myocardium. Implantation into heart tissues of autologous skeletal myoblasts has been associated with improved contractile function in animal models and in humans with acute myocardial ischemia. Since heart infarction is most prevalent in individuals of over 40 years of age, we tested whether culture methods available in our laboratory were adequate to obtain sufficient numbers of differentiated skeletal myoblasts from muscle biopsy specimens obtained from patients aged 41 to 91. METHODS AND RESULTS: No matter of donor age, differentiated skeletal muscle cells could be produced in vitro in amounts adequate for cellular therapy (>/=300 millions). Using desmin as a cytoplasmic marker, about 50% cultured cells were differentiated along myogenic lineages and expressed proteins proper of skeletal muscle (myosin type I and II, actin, actinin, spectrin and dystrophin). Cytogenetic alterations were not detected in cultured muscle cells that had undergone at least 10 population doublings. Molecular methods employed for the screening of persistent viral infections evidenced that HCV failed to replicate in muscle cells cultured from one patient with chronic HCV infection. CONCLUSION: The proposed culture methods appear to hold promise for aged patients not only in the field of cardiovascular medicine, but also in the urologic and orthopedic fields. PMID- 15890078 TI - Race and satisfaction in general OB/GYN clinics. AB - BACKGROUND: The purpose of this study was to test the hypothesis that racial differences in satisfaction can be found among OB/GYN patients on a US naval base. METHODS: Cross-sectional surveys assessing satisfaction with services were obtained from 838 patients who were seen in one of the two general OB/GYN clinics (455 in the base hospital clinic and 391 in a satellite clinic). Multiple logistic regression analysis was used to identify subgroups of patients who were not very satisfied with care received. RESULTS: When the patients seen in the base hospital were analyzed separately, Asian respondents had significantly lower odds of being very satisfied relative to non-Hispanic white respondents (AOR = .33, p < .01). CONCLUSION: Asian patients may be less satisfied than others when treated at a larger facility. Patients treated at a satellite clinic tended to be more satisfied than patients seen at the base hospital. PMID- 15890079 TI - Lifetime environmental tobacco smoke exposure and the risk of chronic obstructive pulmonary disease. AB - BACKGROUND: Exposure to environmental tobacco smoke (ETS), which contains potent respiratory irritants, may lead to chronic airway inflammation and obstruction. Although ETS exposure appears to cause asthma in children and adults, its role in causing COPD has received limited attention in epidemiologic studies. METHODS: Using data from a population-based sample of 2,113 U.S. adults aged 55 to 75 years, we examined the association between lifetime ETS exposure and the risk of developing COPD. Participants were recruited from all 48 contiguous U.S. states by random digit dialing. Lifetime ETS exposure was ascertained by structured telephone interview. We used a standard epidemiologic approach to define COPD based on a self-reported physician diagnosis of chronic bronchitis, emphysema, or COPD. RESULTS: Higher cumulative lifetime home and work exposure were associated with a greater risk of COPD. The highest quartile of lifetime home ETS exposure was associated with a greater risk of COPD, controlling for age, sex, race, personal smoking history, educational attainment, marital status, and occupational exposure to vapors, gas, dusts, or fumes during the longest held job (OR 1.55; 95% CI 1.09 to 2.21). The highest quartile of lifetime workplace ETS exposure was also related to a greater risk of COPD (OR 1.36; 95% CI 1.002 to 1.84). The population attributable fraction was 11% for the highest quartile of home ETS exposure and 7% for work exposure. CONCLUSION: ETS exposure may be an important cause of COPD. Consequently, public policies aimed at preventing public smoking may reduce the burden of COPD-related death and disability, both by reducing direct smoking and ETS exposure. PMID- 15890080 TI - Visualization methods for statistical analysis of microarray clusters. AB - BACKGROUND: The most common method of identifying groups of functionally related genes in microarray data is to apply a clustering algorithm. However, it is impossible to determine which clustering algorithm is most appropriate to apply, and it is difficult to verify the results of any algorithm due to the lack of a gold-standard. Appropriate data visualization tools can aid this analysis process, but existing visualization methods do not specifically address this issue. RESULTS: We present several visualization techniques that incorporate meaningful statistics that are noise-robust for the purpose of analyzing the results of clustering algorithms on microarray data. This includes a rank-based visualization method that is more robust to noise, a difference display method to aid assessments of cluster quality and detection of outliers, and a projection of high dimensional data into a three dimensional space in order to examine relationships between clusters. Our methods are interactive and are dynamically linked together for comprehensive analysis. Further, our approach applies to both protein and gene expression microarrays, and our architecture is scalable for use on both desktop/laptop screens and large-scale display devices. This methodology is implemented in GeneVAnD (Genomic Visual ANalysis of Datasets) and is available at http://function.princeton.edu/GeneVAnD. CONCLUSION: Incorporating relevant statistical information into data visualizations is key for analysis of large biological datasets, particularly because of high levels of noise and the lack of a gold-standard for comparisons. We developed several new visualization techniques and demonstrated their effectiveness for evaluating cluster quality and relationships between clusters. PMID- 15890081 TI - Development and characterization of positively selected brain-adapted SIV. AB - HIV is found in the brains of most infected individuals but only 30% develop neurological disease. Both viral and host factors are thought to contribute to the motor and cognitive disorders resulting from HIV infection. Here, using the SIV/rhesus monkey system, we characterize the salient characteristics of the virus from the brain of animals with neuropathological disorders. Nine unique molecular clones of SIV were derived from virus released by microglia cultured from the brains of two macaques with SIV encephalitis. Sequence analysis revealed a remarkably high level of similarity between their env and nef genes as well as their 3' LTR. As this genotype was found in the brains of two separate animals, and it encoded a set of distinct amino acid changes from the infecting virus, it demonstrates the convergent evolution of the virus to a unique brain-adapted genotype. This genotype was distinct from other macrophage-tropic and neurovirulent strains of SIV. Functional characterization of virus derived from representative clones showed a robust in vitro infection of 174xCEM cells, primary macrophages and primary microglia. The infectious phenotype of this virus is distinct from that shown by other strains of SIV, potentially reflecting the method by which the virus successfully infiltrates and infects the CNS. Positive in vivo selection of a brain-adapted strain of SIV resulted in a near-homogeneous strain of virus with distinct properties that may give clues to the viral basis of neuroAIDS. PMID- 15890082 TI - Predicting functional sites with an automated algorithm suitable for heterogeneous datasets. AB - BACKGROUND: In a previous report (La et al., Proteins, 2005), we have demonstrated that the identification of phylogenetic motifs, protein sequence fragments conserving the overall familial phylogeny, represent a promising approach for sequence/function annotation. Across a structurally and functionally heterogeneous dataset, phylogenetic motifs have been demonstrated to correspond to a wide variety of functional site archetypes, including those defined by surface loops, active site clefts, and less exposed regions. However, in our original demonstration of the technique, phylogenetic motif identification is dependent upon a manually determined similarity threshold, prohibiting large scale application of the technique. RESULTS: In this report, we present an algorithmic approach that determines thresholds without human subjectivity. The approach relies on significant raw data preprocessing to improve signal detection. Subsequently, Partition Around Medoids Clustering (PAMC) of the similarity scores assesses sequence fragments where functional annotation remains in question. The accuracy of the approach is confirmed through comparisons to our previous (manual) results and structural analyses. Triosephosphate isomerase and arginyl-tRNA synthetase are discussed as exemplar cases. A quantitative functional site prediction assessment algorithm indicates that the phylogenetic motif predictions, which require sequence information only, are nearly as good as those from evolutionary trace methods that do incorporate structure. CONCLUSION: The automated threshold detection algorithm has been incorporated into MINER, our web-based phylogenetic motif identification server. MINER is freely available on the web at http://www.pmap.csupomona.edu/MINER/. Pre-calculated functional site predictions of the COG database and an implementation of the threshold detection algorithm, in the R statistical language, can also be accessed at the website. PMID- 15890083 TI - Pathogenesis of Wegener's granulomatosis: current concepts. AB - Wegener's granulomatosis (WG) is a complex autoimmune syndrome that is characterised by upper/lower respiratory necrotising granulomatosis, glomerulonephritis and small-vessel vasculitis. Since Wegener's 1936 description, considerable advances in recognition and treatment have changed this disease from a rapidly and uniformly fatal illness to a chronic disease characterised by remissions and relapses. The serendipitous discovery of anti-neutrophil cytoplasmic antibodies (ANCAs) as a marker associated with WG focused attention on the potential pathogenic role of these antibodies and has recently led to the development of novel animal models that might facilitate our understanding of the disease pathogenesis. Future animal models of this disease will have to account for the role of both ANCA-mediated pathology and granulomatous inflammation to enable us to understand the chronic and persistent features of WG in humans. PMID- 15890084 TI - Does chronic experimental head-down tilt alter intramural innervation density of limb blood vessels? AB - Earlier, substantial increases in the intramural sympathetic innervation density of rat hind-limb blood vessels were found after 2 weeks of experimental orthostasis with tubular 45 degrees head-up tilt cages. In the present study, we presumed that chronic head-down tilting induces opposite changes in the innervation density. Tilted rats were kept 45 degrees head-down in long tubular cages for either 2 or 4 weeks (HDT2, HDT4), and the control animals were maintained in horizontal tilt cages for the same period (HOR2, HOR4). Segments of the saphenous and brachial veins and arteries were used for quantitative electron microscopic examinations. Intramural innervation density was defined by nerve terminal density (NTD) and synaptic microvesicle count (SVC) within the vascular adventitia. Neither HDT2 nor HDT4 resulted in a decrease of NTD or SVC of the saphenous and brachial veins or arteries; instead, a tendency to increase was observed in some cases. Thus in contrast to the large increases we found earlier in hind-limb vascular innervation density after 2 weeks of head-up tilting, head down tilting of the same duration-or even twice as long-did not decrease the adventitial innervation density in our model. We assume that the quasi-free locomotor exercise the tilted animals in the long tubular cages were allowed may counteract a possible suppressive effect of chronic head-down tilt on hind-limb vascular innervation density. PMID- 15890089 TI - [Correlation of E-cadherin polymorphisms to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma]. AB - BACKGROUND & OBJECTIVE: E-cadherin (CDH1) relates with invasion and metastasis of various cancers. Polymorphisms in the promoter region of E-cadherin gene may modify its transcriptional activity and protein level. This study was designed to investigate the correlation of CDH1 C-160A and G-347GA single nucleotide polymorphisms(SNPs) to susceptibilities and lymphatic metastases of esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in northern China population. METHODS: CDH1 promoter SNPs (C-160A and G-347GA) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 333 ESCC patients, 239 GCA patients, and 343 healthy controls. The combined effect of C-160A and G-347GA was analyzed by EH software. RESULTS: The overall genotype and allelotype distributions of C-160A and G-347GA in ESCC and GCA patients were not significantly different from those in healthy controls (P = 0.08). When stratified by smoking status, family history of upper gastrointestinal cancer, and lymph node metastasis state, CDH1 SNPs also did not significantly influence the development and lymphatic metastasis of ESCC and GCA. However, compared with individuals with G-347GA G/G genotype, individuals with GA allele (G/GA or GA/GA genotype) had significantly higher risk to develop GCA [age and gender adjusted odds ratio (OR) = 1.45, 95% confidence interval (CI) = 1.03 2.04]. The haplotype distribution of CDH1 in the 333 ESCC patients was significantly different from that in the 343 healthy controls (P = 0.008). Compared with -160C/-347G haplotype, -160A/-347GA haplotype significantly increased the risk of developing ESCC (age and gender adjusted OR = 24.26, 95% CI = 3.25-180.87). CONCLUSIONS: CDH1 C-160A SNP has no relation with susceptibility and lymphatic metastasis of ESCC and GCA. However, individuals with G-347GA GA allele have high risk of developing GCAu individuals with -160A/-347GA haplotype have high risk of developing ESCC. PMID- 15890090 TI - Inhibitory effect of anti-fas ribozyme on apoptosis of mouse T cells. AB - BACKGROUND & OBJECTIVE: Tumor cells can express FasL, and induce apoptosis of activated T cells expressing Fas, which is so called "Fas counterattack". This study was to investigate inhibitory effect of anti-Fas ribozyme on apoptosis of mouse T cells, and explore a new way to enhance antitumor ability of T cells. METHODS: A hammerhead ribozyme targeting Fas mRNA was synthesized, its in vitro cleavage reaction was performed. Anti-Fas ribozyme was transfected into mouse spleen T cells by electroperation (ribozyme-transfected group), empty control and mock-transfected groups were set up. Fas expression on T cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. After treatment of anti-Fas antibody (JO(2)), cell apoptosis was measured by flow cytometry, proliferation of T cells was measured by MTT assay. In vitro killing activity of cytotoxic T lymphocytes (CTLs) was detected by lactate dehydrogenase (LDH) releasing assay. RESULTS: Anti-Fas ribozyme cleaved Fas mRNA efficiently with a rate of 60%. mRNA level of Fas was significantly lower on ribozyme transfected cells than on control and mock-transfected cells (0.4 vs. 1.1, and 1.0, P < 0.01)u its protein level was according to this result. After treatment of JO(2), cell apoptosis rate was significantly lower in ribozyme-transfected group than in the rest 2 groups (35% vs. 86%, and 87%, P < 0.01), but cell proliferation rate was significantly higher in ribozyme-transfected group than in the rest 2 groups (208% vs. 100%, and 98%, P < 0.01). In vitro killing activity of CTLs was significantly stronger in ribozyme-transfected group than in the rest 2 groups (67% vs. 32%, and 31%, P < 0.01). CONCLUSION: Anti-Fas ribozyme can remarkably suppress Fas expression on mouse activated spleen T cells, and protect T cells from Fas-mediated apoptosis, which may enhance antitumor ability of T cells. PMID- 15890091 TI - [Inhibiting expression of human telomerase reverse transcriptase promotes degradation of survivin protein]. AB - BACKGROUND & OBJECTIVE: The expression of Survivin in cancer cells highly correlates with that of human telomerase reverse transcriptase (hTERT). Both of them are ideal targets for cancer gene therapy. This study aimed to clarify if they regulate each other in cancer cells. METHODS: The expressions of Survivin and hTERT in HeLa S3 cells were inhibited by antisense oligonucleotide respectively. Activity of telomerase was detected by telomerase repeat amplification (TRAP) assay. Protein and mRNA levels of Survivin were analyzed by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) respectively. Proliferation of HeLa S3 cells was analyzed by MTT assay. RESULTS: Inhibiting the expression of Survivin in HeLa S3 cells had no effects on telomerase activity. Inhibiting the expression of hTERT by antisense oligonucleotide No.14 decreased protein level of Survivin, which was negatively correlated with the concentration of No.14 (200-1 000 nmol/L), but didn't change mRNA level of survivin. The decrease of Survivin level was inhibited by proteasome inhibitor lactacystin and MG132. Furthermore, simultaneous inhibition of hTERT and survivin co-efficiently inhibited proliferation of HeLa S3 cells. CONCLUSION: Inhibiting the expression of hTERT in HeLa S3 cells promotes ubiquitin-proteasome degradation of Survivin. PMID- 15890092 TI - [Effect of silencing HIF-1alpha by RNA interference on expression of vascular endothelial growth factor in osteosarcoma cell line SaOS-2 under hypoxia]. AB - BACKGROUND & OBJECTIVE: Hypoxia-inducible factor-1 alpha (HIF-1alpha) is a key regulator for hypoxia tolerance and angiogenesis of tumor. This study was to investigate the expression of HIF-1alpha and vascular endothelial growth factor (VEGF) in human osteosarcoma cell line SaOS-2 under hypoxia, to explore the effect of HIF-1alpha on hypoxia-activated angiogenesis regulation pathway in osteosarcoma. METHODS: CoCl2 was used as chemical hypoxia-inducing reagent to mimic tumor hypoxic microenvironment. mRNA and protein levels of HIF-1alpha and VEGF at different hypoxic culture phases were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. Small hairpin RNAs (shRNAs) eukaryotic expression vector targeting HIF-1alpha was constructed, and transfected into SaOS-2 cells. Western blot was used to detect gene silencing effect on HIF-1alpha. RT-PCR and enzyme linked immunosorbent assay (ELISA) were used to observe the change of VEGF gene expression after HIF-1alpha gene silence. RESULTS: Under hypoxia, mRNA level of HIF-1alpha kept stable, while its protein level increased obviouslyu both mRNA and protein levels of VEGF were up-regulated. The shRNAs plasmid targeting HIF 1alpha gene was constructed successfully, and down-regulated HIF-1alpha gene in SaOS-2 cells efficiently followed by VEGF gene down-regulation. CONCLUSIONS: Hypoxia can increase protein level of HIF-1alpha in osteosarcoma. HIF-1alpha up regulates the gene expression of VEGF via transcription activation which promotes angiogenesis in osteosarcoma under hypoxic microenvironment. PMID- 15890093 TI - [Killing effect of Ad.TERT-TRAIL on tumor cell lines and its mechanism]. AB - BACKGROUND & OBJECTIVE: Oncolytic adenovirus Ad.TERT, a novel tumor-specific proliferating virus, has been constructed by replacing normal promoter of mild type adenovirus E1A with promoter of human telomerase reverse transcriptase (hTERT). In vitro and in vivo experiments confirmed that Ad.TERT has antitumor effect. This study was to construct Ad.TERT-TRAIL through inserting apoptosis gene trail into Ad.TERT, and explore its antitumor effect and mechanism. METHODS: Plasmid pZhTERT-trail and adenovirus packaging plasmid pBHGE3 were homologously recombined in HEK293 cells to construct Ad.TERT-TRAIL. Ad.TERT-TRAIL was identified by polymerase chain reaction (PCR), and confirmed by Western blot. Killing effects of Ad.TERT-TRAIL and Ad.TERT on 3 tumor cell lines, SW620, BEL 7404 and Bcap-37, and a normal cell line NHLF were detected by crystal violet dye method or MTT assay. Expression of Caspase-3 in Ad.TERT-TRAIL-, and Ad.TERT transfected SW620 cells was detected by Western blotu cell apoptosis was detected by flow cytometry (FCM). RESULTS: The 860 bp-length trail gene has been amplified by PCR. Western blot showed trail and E1A only expressed in tumor cells, which confirmed the successful construction of Ad.TERT-TRAIL. Killing effects of Ad.TERT-TRAIL on tumor cells were 10-100 times as strong as that of Ad.TERTu while both of them had little effects on normal cells. After 3 days infection (100 multiple of infection, MOI), survival rate of Ad.TERT-TRAIL-infected SW620 cells was 4%, but that of Ad.TERT-infected SW620 cells was 56%u both viruses had little effects on NHLF cells. Expression of Caspase-3 was higher in Ad.TERT-TRAIL infected SW620 cells than in Ad.TERT-infected SW620 cells. Apoptosis rate of Ad.TERT-TRAIL-infected SW620 cells was 4 times as high as that of Ad.TERT infected SW620 cells. CONCLUSIONS: Ad.TERT-TRAIL has much stronger antitumor effect than Ad.TERT. Its effect might relate with inducement effects of trail gene on expression of Caspase-3, and apoptosis of tumor cells. PMID- 15890094 TI - [Expression of band 3 protein on erythrocytes of malignant tumor patients and its impact on proliferation of K562 cells]. AB - BACKGROUND & OBJECTIVE: Membrane domain of band 3 protein (mdb3) mediates transmembrane exchange of chloride and bicarbonate, and regulates intracellular pH. It has been found recently that abnormality of CI(-)/HCO3(-) exchange, which mainly leads to change of intracellular pH, may be involved in cell proliferation and apoptosis. This study was to explore expression of band 3 protein on erythrocytes, and its impact on proliferation of K562 cells. METHODS: Anion transport activity of band 3 protein on erythrocytes of 8 malignant tumor patients was measured using SPQ fluorescent probe. Expression of band 3 protein was detected by Western blot. Plasmid pYD1-mdb3 was constructed, and transfected into K562 cells. Cl- transport activity and proliferation of K562 cells were detected after transfection. RESULTS: Of the 8 patients, 7 showed increase of anion transport activity on erythrocytes, 5 showed increase of band 3 protein expression. To some extent, expression of mdb3 enhanced proliferation of K562 cells. CONCLUSION: Expression of band 3 protein is enhanced on erythrocytes of some malignant tumors, and might be a candidate marker of malignant tumors. PMID- 15890095 TI - [Specific anti-glioma angiogenesis immune response induced by attenuated Salmonella typhimurium vaccine expressing vascular endothelial growth factor receptor-2]. AB - BACKGROUND & OBJECTIVE: Vascular endothelial growth factor (VEGF) and its receptor (VEGFR-2) play important roles in tumor angiogenesis. This study was to investigate anti-vasculature and anti-glioma effects of attenuated Salmonella typhimurium vaccine expressing VEGFR2 gene. METHODS: Plasmid pcDNA3.1-VEGFR2 was constructed, and electrotransfected into attenuated Salmonella typhimurium strain SL7207. C57BL/6J mice were immunized with gastrogavage of cDNA vaccine encoding VEGFR2 (vaccine group). C57BL/6J mice received gastrogavage of pcDNA3.1 or NaHCO3 were used as controls. Serum level of specific anti-VEGFR2-IgG antibody was detected by ELISA. Cytotoxic T lymphocytes (CTLs) activity was measured by MTT assay. Mouse models of intracranial Gl261 glioblastoma were treated with gastrogavage of attenuated Salmonella typhimurium expressing VEGFR2 gene. Tumor diameter was measuredu microvessel density (MVD) was detected by immunohistochemistryu tumor cell apoptosis was detected by TUNEL. RESULTS: All mice immunized with the vaccine developed high levels of anti-VEGFR2-IgG antibody, and showed strong CTLs activities against VEGFR2. The vaccine substantially inhibited glioblastoma growth. MVD was significantly lower in vaccine group than in pcDNA3.1 group, and NaHCO3 group (8.8+/-1.9 vs. 27.2+/-4.5, and 26.5+/-5.8, P < 0.01)u while apoptotic cell count per visual field was significantly higher in vaccine group than in the rest 2 groups (23.4+/-4.7 vs. 3.1+/-1.0, and 4.4+/-1.2, P < 0.01). CONCLUSION: Attenuated Salmonella typhimurium vaccine expressing VEGFR2 gene can break immunologic tolerance against self-VEGFR2 antigen, and specifically kill glioblastoma vascular endothelial cells by inducing specific anti-VEGFR2 immunoreaction. PMID- 15890096 TI - [Cell balance between apoptosis and proliferation in colon cancer and its correlation with prognosis]. AB - BACKGROUND & OBJECTIVE: In normal tissues and organs, cell apoptosis and proliferation maintain a homeostasis. Alterations of this physiologic balance may lead to malignant transformation of cells and tumorigenesis. This study was to investigate cell balance (ratio of apoptosis index to proliferation index, AI/PI) in colon cancer and its correlation with prognosis. METHODS: The apoptotic population and the proliferating population of colon cancer cells were quantitatively analyzed by Sub-G1 method and Ki-67/DNA bivariate analysis of flow cytometry. Cell proliferation was observed under confocal microscope. Kaplan Meier method and log-rank test were used to analyze patients' survival. RESULTS: AI/PI ratio of cells was significantly higher in normal colon tissue than in colon adenoma tissue, Dukes' A colon cancer, Dukes' B colon cancer, and Dukes' C and D colon cancer (0.45+/-0.19 vs. 0.30+/-0.07, 0.29+/-0.11, 0.28+/-0.10, and 0.26+/-0.07, respectively, P < 0.01). AI/PI ratio showed a down-regulating trend in variables tested including tumor size, pathologic type, differentiation grade, Dukes' stage, and nodal involvement. AI/PI ratio of peripheral lymphocytes was significantly higher in colon cancer patients than in healthy people (0.64+/-0.11 vs. 0.49+/-0.12, P < 0.01). The expression of Ki-67 was observed in normal colon tissue, colon adenoma, and colon cancer under confocal microscope. Survival rate of patients with AI/PI ratio of < 0.285 (using the median value as the cutoff) and >/= 0.285 was not significantly different (P> 0.05). CONCLUSIONS: Dysregulation (down-regulation or up-regulation) of cell balance between apoptosis and proliferation in colon cancer cells and lymphocytes might play an important role in tumorigenesis and tumor progression. AI/PI ratio can' t be used as a prognostic factor of colon cancer. PMID- 15890097 TI - [Anti-pancreatic cancer immune response induced by K-ras mutated peptide]. AB - BACKGROUND & OBJECTIVE: Since most tumor antigens are uncertain, the immunotherapy against tumors still lingers in clinical trials. Because k-ras proto-oncogene specifically expresses in pancreatic cancer cells with constant mutation site, it may be an ideal target for immunotherapy against pancreatic cancer. This study was to evaluate the feasibility of inducing immunotherapy on pancreatic cancer by K-ras mutated peptides, give experimental evidence to clinical individual therapy on pancreatic cancer. METHODS: k-ras oncogene was amplified by reverse transcription-polymerase chain reaction (RT-PCR)u its mutation type was detected by flow cytometry. T cells were induced by dendritic cells, which had been pulsed with synthesized mutated peptide. Killing effect of tumor specific T cells on pancreatic cancer cell line Patu8988 was assessed by MTT assay. RESULTS: There was a point mutation (GGT-->GTT) in the 12th codon in Patu8988 cellu the amino acid was mutated to valine. The mutation epitope was efficiently presented on dendritic cells'surface. The cytotoxic T cells induced by mutated peptide could kill Patu8988 cells efficiently. CONCLUSIONS: The mutated peptide can efficiently induce immunocytes to kill pancreatic cancer cells, which have k-ras mutation site. This finding provides experimental clue for immunotherapy against pancreatic cancer. PMID- 15890098 TI - [Risk factors of gastric cancer in Wuwei City---an endemic region of gastric cancer]. AB - BACKGROUND & OBJECTIVE: Researches show that epidemiologic factors of gastric cancer include living habit, eating moldy food and pickles, dystrophy, lack of microelements, and inherit, etc. This study was to explore universalities of these factors in Wuwei, a city in northwest China with high incidence of gastric cancer, and provide evidences for the first-prevention of gastric cancer. METHODS: Family histories of the residents in Wuwei City were investigated with case-control method. Separating ratio and heredity degree of gastric cancer were calculated with Li-Mantel-Cart method and Falconer's regression method. Chronic gastritis patients were followed-up by home-visit, gastroscopy, and pathology. Cancerigenic fungi and volatility N-nitrosate compounds in residents' meal, Helicobacter pylori (Hp) in gastric mucosa, and total content of vitamin C in 293 healthy adults' serum were detected by culture, authentication, and laboratory examinations. RESULTS: In Wuwei City, the separating ratio of gastric cancer was 0.077; the heredity degree of first-degree relatives was 22.91%u cancerization rate of year in person of atrophic gastritis crowd was 1.09%. Eight kinds of nitrosamine and 14 kinds of cancerigenic fungi were detected from residents' meal. Total content of vitamin C in serum of the 293 healthy adults in summer was (5.74+2.79) mg/L. Positive rate of Hp in gastric mucosa of the residents was 67%. CONCLUSION: The major extrinsic factors of gastric cancer in Wuwei City include various strong carcinogens existing in residents' meal and lack of vitamin Cu its intrinsic factors include infection of Hp, atrophic gastritis (especially atypical hyperplasia), and heritage susceptibility. PMID- 15890099 TI - [Correlation of expression of connexin to growth and progression of cervical carcinoma in situ]. AB - BACKGROUND & OBJECTIVE: Gap junction intercellular communication (GJIC), mediated by connexin (CX), is the unique type of intercellular communication in carcinoma in situ (CIS). Changes in expression of CXs and function of GJIC may play roles in carcinogenesis of cervical cancer and progression of CIS. This study was to investigate the expression of CXs in normal epithelium (NE), hyperplasia, CIS, and invasive carcinoma (IC) of cervix, to explore correlation of expression of CXs to pathogenesis and progression of cervical CIS. METHODS: Expression of CX43 (mRNA, protein), CX26 (mRNA), and Ki-67 (protein) in 30 specimens of cervical NE, 22 specimens of cervical hyperplasia, 30 specimens of cervical CIS, and 26 specimens of cervical IC were detected by immunohistochemistry and in situ hybridization. RESULTS: In cervical NE, hyperplasia, CIS, and IC, positive rates of CX43 protein were 83%, 55%, 50%, and 30%, respectively, and positive rates of CX43 mRNA were 97%, 64%, 58%, and 46%, respectively; positive rates of CX26 mRNA was 93%, 68%, 55%, and 42%, respectively. Positive rates of CX43 (mRNA, protein) and CX26 (mRNA) were gradually decreased. Positive rate of CX was significantly lower in CIS group than in NE group (P < 0.05). The expressions of CX26 and CX43 were weaker in CIS group than in NE groupu their expressions were obviously decreased, or even vanished in the area of CIS adjacent to NE. Proliferating index (PI), indicated by expression of Ki-67 protein, was increased gradually in NE (5%), hyperplasia (12%), CIS (30%), and IC (62%)u the differences were significant (P < 0.05) between CIS group and other groups. CONCLUSIONS: CX26 and CX43 may play important roles in carcinogenesis of cervical cancer. CXs may inhibit tumorigenesis and cell proliferation; it might be key factors in earlier stages when cellular malignant phenotypes have been established, or NE has been transformed into CIS. PMID- 15890100 TI - [Eukaryotic expression and intracellular distribution of cancer and embryo expression protein 65]. AB - BACKGROUND & OBJECTIVE: Previous studies show that cancer and embryo expression protein 65 (CEP65) only expresses in various cancer cells and embryo cells. This study was to detect intracellular distribution of CEP65, and make the foundation for further CEP65 function study. METHODS: Two kinds of eukaryotic expression plasmids, which can separately express full-length CEP65-GST and fluorescent fusion protein CEP65-Red, were constructed, and transfected into COS7 cells by DEAE-Dextran method. Plasmosin and nucleoprotein of COS7 cells were separated. Intracellular distribution of CEP65 was detected by Western blot, and observed under fluorescent microscope. RESULTS: Western blot confirmed that CEP65 distributed in cytoplasm and nuclei. Fluorescent protein localization showed diffuse distribution of fluorescence in the cells transfected with pcDNA3-Red, and collective fluorescent articles in cytoplasm and nuclei of the cells transfected with pcDNA3-Red-Cep65. CONCLUSIONS: CEP65 can express efficiently in COS7 cells, and locates in cytoplasm and nuclei. It may be a kind of nucleus associated protein. PMID- 15890101 TI - [Primary effect of submandibular salivary gland transfer in preventing radiation induced xerostomia of nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Xerostomia, an ubiquitous sequelae in nasopharyngeal carcinoma (NPC) patients after radiotherapy, seriously affects life quality of the patients. This study was to investigate clinical value of submandibular salivary gland transfer in preventing radiation-induced xerostomia of NPC patients. METHODS: A total of 70 eligible patients with NPC were recruited, and divided into test group (36 patients) and control group (34 patients). In test group, the submandibular glands were transferred to the submental spaces before conventional radiotherapy. The submental spaces were shielded during radiotherapy. Oral cavity mucous membrane reaction during radiotherapy, submandibular gland function and salivary fluid before and after radiotherapy, and questionnaire of xerostomia 3 months after radiotherapy were compared between test group and control group. RESULTS: Acute oral cavity mucous membrane reaction was severer in control group than in test group; the incidence of stage III reaction was significantly higher in control group than in test group ( 8/34 vs. 3/36, P = 0.007). Three months after radiotherapy, trapping and excretion functions of the transferred submandibular glands were significantly better than those of the untransferred submandibular glands in test group (P = 0.001 and P = 0.000); trapping and excretion functions of the submandibular glands were significantly better in test group than in control group (P = 0.004 and P = 0.000). The mean weight of salivary fluid after radiotherapy was heavier in test group than in control group (1.41 g vs. 0.71 g). Incidence of moderate to severe degree of xerostomia was significantly higher in test group than in control group (76.5% vs. 13.9%, P = 0.000). CONCLUSION: Submandibular gland transfer may be used to prevent radiation-induced xerostomia of NPC patients, and may improve life quality of the patients. PMID- 15890102 TI - [Phase I study of CM-Na combined with concurrent radiochemotherapy for advanced esophageal carcinoma]. AB - BACKGROUND & OBJECTIVE: Although concurrent radiochemotherapy is recommended as the standard treatment for advanced esophageal carcinoma, the local failure still reaches up to 44%-54%. This study aimed to explore maximum tolerance dose (MTD) of sodium glycididazole(CM-Na) in the combined planning as the recommended dose for phase II study. METHODS: Twenty-two patients with pathologically confirmed esophageal squamous carcinoma of stage III-IV were recruited according to the inclusion criteria. All patients were divided into 4 groups (at least 3 patients in a group) by turn as the dose of CM-Na escalated from 400 to 600, 700, and 800 mg x (m(2) x d)(-1) by Fibonacci's method, and treated according to the plan. All patients underwent the same concurrent radiochemotherapy. Conventional radiotherapy was performed with total dose of 60 Gy within 6 weeks. CM-Na was given 1 h before radiotherapy at Monday, Wednesday, and Friday every week, combined with 2 circles of continuous 5-day chemotherapy of 500 mg x (m(2) x d)( 1) of 5-fluoruracil (5-FU) and 20 mg x (m(2) x d)(-1) of cisplatin (DDP) at the first and the fifth week. RESULTS: Low-grade gastrointestinal adverse reactions were observed in the 4 groups during the period of chemotherapy, but no adverse reactions of nervous system, kidney, or heart were observed. Severe adverse reactions occurred in 800 mg x (m(2) x d) (-1) group, included 3 cases of grade III radioactive esophagitis, 2 cases of grade IV aminopherase risen, and 1 case of grade III thrombocytopenia. All the 19 patients in the rest 3 groups suffered grade I-II thrombocytopenia. The rates of complete remission and partial remission were 27% (6/22) and 68% (15/22) at the end of treating, and 46% (10/22) and 54% (12/22) 1 month after treating. CONCLUSION: Liver disfunction is the main dose-limited toxicity of the treatment schemeu 700 mg x (m(2) x d) (-1) of CM-Na is recommended to phase II clinical study. PMID- 15890103 TI - [Treating renal carcinoma with ablation or ablation combined with 125I seed implantation: a report of eleven cases]. AB - BACKGROUND & OBJECTIVE: Recently, multi-needles radiofrequency ablation and brachyradiotherapy have been used to treat some solid tumors, such as liver cancer and prostate cancer. This study was to explore the feasibility and efficacy of radiofrequency ablation or ablation combined with 125I seed implantation on treating renal carcinoma. METHODS: From Jun. 2000 to Feb. 2004, 11 patients with a total of 13 renal carcinoma lesions were treated with ablation or ablation combined with (125)I seed implantation, including 6 cases of renal carcinoma of solitary kidney, 2 cases of renal carcinoma combined with contralateral renal atrophy resulted from ureteric stone, 1 case of bilateral tumor, and 2 cases with general conditions contraindicated to surgery. Of the 13 lesions, 1 was resected by open operation, 6 were treated with multi-needles radiofrequency ablation, and 6 were treated with ablation and simultaneous (125)I seed implantation around tumor margin. RESULTS: The patients were followed-up for 6-46 months (mean 27 months). One patient died of cardiac muscle infarction 4 months after operation. Of the 10 alive patients, 2 with recurrent tumors 7 months and 13 months after operation were treated with ablation again, 1 with postoperative uremia was treated with intermittent peritoneal dialysis, the rest 7 had normal renal function. CONCLUSION: Multi-needles radiofrequency ablation or ablation combined with (125)I seed implantation would be an option in the treatment for patients with renal carcinoma of solitary kidney or bilateral kidney lesions, or for patients can't suffer from surgery. PMID- 15890104 TI - [Magnetic resonance imaging diagnosis of occupied lesions in cerebellopontine angle area:a report of 78 cases]. AB - BACKGROUND & OBJECTIVE: It is difficult to diagnose cerebellopontine angle area tumor because of many sorts and origins of the tumor. This study was to explore magnetic resonance imaging (MRI) diagnosis of occupied lesions in cerebellopontine angle area. METHODS: MRI records of 78 patients with pathologically confirmed occupied lesions in cerebellopontine angle area were analyzed. RESULTS: Of the 78 cases, 48 (61.5%) were unilateral acoustic neuroma, 5(6.4%) were bilateral acoustic neuroma, 12 (15.4%) were meningioma, 4 (5.1%) were trigeminal neuroma, 3 (3.8%) were hemangioblastoma, 3 (3.8%) were lipoma, 2 (2.6%) were melanoma, 1 (1.3%) was medulloblastoma. According to the anatomic site, tumor lesion character, and MRI signal character, the majority of cerebellopontine angle area tumors were diagnosed accurately. CONCLUSION: MRI plays an important role in diagnosis of occupied lesions in cerebellopontine angle area. PMID- 15890105 TI - [Multivariate prognostic analysis in gastric carcinoma patients after radical operation]. AB - BACKGROUND & OBJECTIVE: Whether received radical operation is an important prognostic factor of gastric carcinoma. But the long-term efficacies of radical operation on different patients are not the same. This study was to investigate prognostic factors of gastric carcinoma. METHODS: Clinical data of 405 patients with gastric carcinoma, received radical operation from Jan. 1985 to Dec. 1995 in Cancer Center of Sun Yat-sen University, were analyzed retrospectively. Life table method was used to analyze survival rate, Wilcoxon test was used for statistical comparison, and Cox regression model was used for multivariate analysis. RESULTS: The 5-year overall survival rate was 43.4%. The 5-year survival rates of patients in pathologic TNM (pTNM) stage I, II, III, and IV were 75.6%, 58.7%, 28.0%, and 18.4%, respectively (P < 0.01). The 5- year survival rates of patients with tumor sizes of less than 2.0 cm, 2.0-3.9 cm, 4.0-5.9 cm, 6.0-7.9 cm, and no less than 8.0 cm were 82.0%, 57.4%, 43.7%, 38.7%, and 26.9%, respectively (P < 0.05). In addition, the 5-year survival rate was higher in patients with perioperative chemotherapy than in patients without perioperative chemotherapy (47.2% vs. 37.8%, P < 0.05). Univariate analysis showed that perioperative chemotherapy, Borrmann type, tumor size, pathologic type, and pTNM stage were prognostic factors of gastric carcinoma. Multivariate analysis showed that pTNM stage, tumor size, and perioperative chemotherapy were independent prognostic factors of gastric carcinoma. CONCLUSIONS: pTNM stage, tumor size, and perioperative chemotherapy are the most significant factors influencing prognosis of gastric carcinoma patients after radical operation. Perioperative chemotherapy contributes to enhance survival rate of gastric carcinoma patients. PMID- 15890106 TI - [Meta-analysis of postoperative adjuvant chemotherapy for Dukes' B colorectal carcinoma]. AB - BACKGROUND & OBJECTIVE: The necessity of adjuvant chemotherapy after radical surgery for patients with Dukes' B (stage II) colorectal carcinoma remains controversial. This study was to evaluate effect of postoperative adjuvant chemotherapy on survival of Dukes' B patients with meta-analysis. METHODS: The results of literatures on postoperative adjuvant chemotherapy for Dukes' B patients from 1985 to 2003 were analyzed synthetically.The 5-year survival rates of postoperative adjuvant chemotherapy group and surgery alone group were compared. RESULTS: Eight published randomized controlled trails were eligible, and had 6 518 patients totally. The 5-year mortality was lower in postoperative adjuvant chemotherapy group than in surgery alone group with odds ratio (OR) of 0.79, and 95% confidence interval (CI) of 0.7-0.9, (P<0.05). CONCLUSION: Postoperative adjuvant chemotherapy could improve 5-year survival rate of Dukes' B patients. PMID- 15890107 TI - [Detecting bone marrow micrometastasis of gastric cancer by magnetic activated cell sorting combined with fluorescent activated cell sorting]. AB - BACKGROUND & OBJECTIVE: Several methods are used to detect bone marrow micrometastasis of gastric cancer with different accuracies. In breast cancer, tumor cells in blood can be detected sensitively and specifically by magnetic activated cell sorting (MACS) and fluorescent activated cell sorting (FACS). This study was to investigate the clinical value of this method in detecting bone marrow micrometastasis of gastric cancer. METHODS: Thirty-five patients, who received operation for gastric cancer from Dec. 2002 to Jun. 2003, were selected. Mononuclear cells were separated from their bone marrows. After marked by MACS minibeads conjugated with cytokeratin (CK) 7/8 antibodies, anti-CK-fluorescein isothiocyanate (FITC), and anti-CD45-perdinin chlorophyll protein (PerCP), tumor cells were enriched twice by MS+/RS+ positive separation column. FACS analysis was conducted on these samples before and after MACS enrichment. The results were compared with clinicopathologic parameters. RESULTS: Disseminated tumor cells were detected in bone marrow of 3 samples(8.6%) before MACS enrichment, and 25 samples (71.4%) after enrichment. The frequencies of tumor cells were 1.4 x 10( 8)-2.4 x 10(-5), 2.2 x 10(-7) -3.7 x 10(-5), and 4.0 x 10-(6)-8.6 x 10(-5) in patients with moderately differentiated, poorly differentiated, and undifferentiated carcinoma, respectively, with significant differences (P = 0.026). Bone marrow micrometastasis positively correlated with tumor TNM stage (P = 0.008), while had no correlation with tumor size, depth of wall invasion, and other clinicopathologic parameters. CONCLUSIONS: MACS combined with FACS may improve detection rate of bone marrow micrometastasis of gastric cancer. The patients with poor differentiation and in advanced TNM stage have more disseminated tumor cells in bone marrow. PMID- 15890108 TI - [Correlation of endothelin A receptor expression to prognosis of nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Endothelin A receptor (ET(A)R) activation contributes to tumor growth and metastasis, including cell proliferation, cell death, angiogenesis, and metastatic spread. In this study, we evaluated correlation of ET(A)R expression to clinical features of nasopharyngeal carcinoma (NPC), and explored its correlation to prognosis. METHODS: Expression of ET(A)R in 153 specimens of NPC was detected by SABC immunohistochemistry. RESULTS: Positive rate of ET(A)R was 73.9% in the 153 NPC specimens. No correlation was found between ET(A)R expression and gender, age, T stage, N stage, and TNM stage of the patients. The 3-year overall survival rate, relapse-free survival rate, and distant metastasis-free survival rate were significantly higher in ET(A)R negative patients than in ET(AR-positive patients (87.5% vs. 73.2%, P = 0.029; 80.0% vs. 57.3%, P = 0.009; and 89.9% vs. 70.4%, P = 0.012), except for local relapse-free survival rate (86.9% vs. 80.2%, P = 0.228). Cox multivariate analysis showed that ET(A)R expression, gender, age, T stage, and N stage were independent prognostic factors, which affected overall survival, relapse-free survival, and distant metastasis. CONCLUSION: ET(A)R expression is an important distant metastasis-related risk factor for patients with NPC; its overexpression indicates poor prognosis. PMID- 15890109 TI - [Expressions of Nm23, E-cadherin, and beta-catenin in non-small cell lung cancer and their correlations with metastasis and prognosis]. AB - BACKGROUND & OBJECTIVE: Recently, studies showed nm23, E-cadherin, and Catenins play important roles in cellular signal transduction, which enhance complexity of their functions in tumor metastasis and can partly explain the diversity of the results from different studies. This study was to investigate correlations of expressions of nm23, E-cadherin, and beta-Catenin in non-small cell lung cancer (NSCLC) to metastasis and prognosis, and their interrelations. METHODS: Expressions of nm23, E-cadherin, and beta-Catenin in 112 specimens of NSCLC and 30 specimens of benign pulmonary lesion were detected by SP immunohistochemistry. RESULTS: The expressions of nm23, E-cadherin and beta-Catenin were significantly weaker in NSCLC tissues than in adjacent non-cancerous tissues, and benign pulmonary tissues (53.0% vs. 64.8%, and 76.9%, P < 0.01; 53.1% vs. 79.7%, and 83.5%, P < 0.01; and 47.2% vs. 80.6%, and 85.6%, P < 0.01), significantly weaker in NSCLC tissues with lymph node metastasis than in those without metastasis (48.0% vs. 65.0%, P < 0.01; 47.3% vs. 60.5%, P < 0.01; and 41.8% vs. 60.3%, P < 0.01), and significantly weaker in NSCLC tissues of stage III-IV than in those of stage I-II (44.8% vs. 67.2%, P < 0.01; 46.6% vs. 64.3%, P < 0.01; 38.1% vs. 63.1%, P < 0.01). The 5-year survival rates of patients with weak expressions of nm23, E-cadherin, and beta-Catenin were significantly lower than those of patients with strong expressions (3.6% vs. 39.3%, P < 0.01; 6.8% vs. 35.8%, P < 0.01; and 3.8% vs. 37.3%, P < 0.01). CONCLUSIONS: Down-regulations of nm23, E cadherin, and beta-catenin closely relate to metastasis of NSCLC. Detection of nm23, E-cadherin, and beta-Catenin might be helpful to predict prognosis of NSCLC patients. PMID- 15890110 TI - [Correlation of NY-ESO-1 gene and protein expression to metastasis and clinicopathologic features of hepatocellular carcinoma]. AB - BACKGROUND & OBJECTIVE: NY-ESO-1 belongs to cancer-testis antigen family. It can inspire both cellular and humoral immune responses in tumor patients, and is regarded as the strongest tumor antigen. This study was to investigate the expression of NY-ESO-1 gene and its correlation with clinicopathologic features of hepatocellular carcinoma (HCC). METHODS: NY-ESO-1 mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) in 62 specimens of HCC and adjacent liver tissue. NY-ESO-1 protein expression and its distribution were detected by immunohistochemistry (IHC) in a tissue microarray contained 132 eligible cases of HCC. RESULTS: Positive rate of NY-ESO-1 mRNA was 27.4% in HCC; it was higher in HCC with tumor embolus of portal vein than in HCC without tumor embolism (40.0% vs. 18.9%). Positive rate of NY-ESO-1 protein was 18.9% in HCC tissue microarray; it was significantly higher in HCC with metastasis than in HCC without metastasis (29.6% vs. 11.5%, P < 0.05). NY-ESO-1 protein mainly located in cytoplasm of HCC cells. Positive rates of NY-ESO-1 mRNA and protein were 28.3% and 19.1% respectively in HBsAg positive HCC, and were 29.5% and 20.7% respectively in HCC with alpha fetoprotein (AFP) of > 20 ng/ml. Both NY-ESO-1 mRNA and protein were not detected in adjacent normal liver tissue. CONCLUSIONS: NY-ESO-1 gene specifically expresses in HCC, and may correlates with progress and metastasis of HCC. It may be a candidate target for antigen-specific immunotherapy for HCC with metastatic lesion. NY-ESO-1 expression has no correlation with HBsAg/AFP status of HCC. PMID- 15890111 TI - [Radiotherapy on neck for nasopharyngeal carcinoma patients with negative cervical lymph node]. AB - BACKGROUND & OBJECTIVE: Nasopharyngeal carcinoma (NPC) patients with negative cervical lymph node (N0-NPC) account for 20% -30% of all NPC patients. Prophylactic irradiation to the upper neck is a criterion of treatment for these patients. But some authors suggest to prophylactically irradiate the the whole neck. This study was to analyze and compare long-term outcomes of the N0-NPC patients treated by prophylactic irradiation to the upper neck and to the whole neck, and explore radiotherapy on neck for N0-NPC patients. METHODS: A total of 178 N0-NPC patients, proved by pathology, were treated in our hospital from Jul. 1998 to Dec. 1999. Of the 178 patients, 88 received prophylactic irradiation to the upper neck (upper neck group), 90 received prophylactic irradiation to the whole neck (whole neck group). Kaplan-Meier method and log-rank test were used to analyze and compare survival rate of the 2 groups. Cox model was used to study prognostic factors of N0-NPC. RESULTS: The 1-, 3-, and 5-year overall survival rates were 98.9%, 90.9%, and 80.7% in upper neck group, and 97.8%, 80.0%, and 71.1% in whole neck group (P = 0.224). The 1-, 3-, and 5-year disease-free survival rates were 93.2%, 80.7%, and 77.3% in upper neck group, and 85.6%, 68.9%, and 64.4% in whole neck group (P = 0.163). The incidence of cervical lymph nodes recurrence was 1.14% in upper neck group, and 1.09% in whole neck group (P> 0.05). Multivariate analysis showed that gender, T stage, and short-term outcome were independent prognostic factors of N0-NPC. CONCLUSION: Prophylactic irradiation to upper neck is recommended for N0-NPC patients. PMID- 15890112 TI - [Construction and preliminary identification of subtracted cDNA library of leukemia cell line K562]. AB - OBJECTIVE: Subtractive hybridization technology is a common method to screen and clone differentially expressed genes. This study was to construct subtracted cDNA library of leukemia cell line K562, and screen for differentially expressed genes. METHODS: cDNA fragments of K562 cells (tester), prepared by restriction display (RD), were subtracted with the Sau3A I-digested cDNA fragments of normal lymphocytes (driver). The subtracted cDNA fragments were re-amplified, and cloned into pMD18-T vectors. Positive clones were selected by blue-white screening. The inserts in plasmid were amplified by polymerase chain reaction (PCR), and some of which were sequenced. RESULTS: The subtracted library contained 360 positive clones with cDNA fragments distributed mainly from 200 to 800 bp. The 50 randomly sequenced clones were derived from 42 known genes. CONCLUSION: Specific subtracted cDNA library of K562 cells was successfully constructed with reliable quality, and may be used to further screen and clone differentially expressed genes of K562 cells. PMID- 15890113 TI - [Immunologic characteristics of tumor-derived vascular endothelial cells]. AB - Tumor vessel plays an important role in development of tumor, because they supply tumor not only nutrition but also a way of metastasis. Since tumor-derived vascular endothelial cells chronically emerge in tumor microenvironment, their phenotypes and functions have significantly changed, including some immunologic characteristics, such as down-regulated adhesion molecules, diminished leukocyte endothelium interactions, impaired antigen presentation, enhanced resistant activity to free radicals, and synthesis of a great amount of extracellular matrix. Because tumor-derived vascular endothelial cell is the first barrier for immune cells and immune drugs to enter tumor tissue, these changes of immunologic characteristics of tumor- derived vascular endothelial cells may be related to the mechanisms of tumor cells escaping from host's immune surveillance and immune killing. This review summarized the immunologic characteristics of tumor-derived vascular endothelial cells, and analyzed the relationship between these characteristics and tumor immune escape. PMID- 15890114 TI - Adenovirus type 7 peptide diversity during outbreak, Korea, 1995-2000. AB - To understand the molecular basis of observed regional shifts in the genome types of adenovirus type 7 (Ad7) isolated in Korea during nationwide outbreaks from 1995 to 2000, the genetic variabilities of Ad7d and Ad7l were studied by sequence analysis of hexon, fiber, E3, and E4 open reading frame (ORF) 6/7 peptides. One amino acid change in the receptor-binding domain of fiber and 6 amino acid variations in E4 ORF 6/7 were identified between 2 genome types, while no variations were found in hexon and E3. Phylogenetic trees based on hexon, fiber, and E4 suggested that the Ad7 epidemic was probably caused by the introduction of the Japanese Ad7d strains. Our data also provide evidence that the rapid divergence of Ad7d to a novel genome type Ad7l could have been due to viral strategies involving multiple sequence changes in E4. This result suggests fiber and E4 ORF 6/7 peptides participate in the evolution of Ad7. PMID- 15890115 TI - Risk factors for kala-azar in Bangladesh. AB - Since 1990, South Asia has experienced a resurgence of kala-azar (visceral leishmaniasis). To determine risk factors for kala-azar, we performed cross sectional surveys over a 3-year period in a Bangladeshi community. By history, active case detection, and serologic screening, 155 of 2,356 residents had kala azar with onset from 2000 to 2003. Risk was highest for persons 3-45 years of age, and no significant difference by sex was seen. In age-adjusted multivariable models, 3 factors were identified: proximity to a previous kala-azar patient (odds ratio [OR] 25.4, 95% confidence interval [CI] 15-44 within household; OR 3.2 95% CI 1.7-6.1 within 50 m), bed net use in summer (OR 0.7, 95% CI 0.53 0.93), and cattle per 1,000 m2 (OR 0.8, 95% CI 0.70-0.94]). No difference was seen by income, education, or occupation; land ownership or other assets; housing materials and condition; or keeping goats or chickens inside bedrooms. Our data confirm strong clustering and suggest that insecticide-treated nets could be effective in preventing kala-azar. PMID- 15890116 TI - Venezuelan equine encephalitis virus infection of spiny rats. AB - Enzootic strains of Venezuelan equine encephalitis virus (VEEV) circulate in forested habitats of Mexico, Central, and South America, and spiny rats (Proechimys spp.) are believed to be the principal reservoir hosts in several foci. To better understand the host-pathogen interactions and resistance to disease characteristic of many reservoir hosts, we performed experimental infections of F1 progeny from Proechimys chrysaeolus collected at a Colombian enzootic VEEV focus using sympatric and allopatric virus strains. All animals became viremic with a mean peak titer of 3.3 log10 PFU/mL, and all seroconverted with antibody titers from 1:20 to 1:640, which persisted up to 15 months. No signs of disease were observed, including after intracerebral injections. The lack of detectable disease and limited histopathologic lesions in these animals contrast dramatically with the severe disease and histopathologic findings observed in other laboratory rodents and humans, and support their role as reservoir hosts with a long-term coevolutionary relationship to VEEV. PMID- 15890117 TI - Assessing parents' perception of children's risk for recreational water illnesses. AB - Understanding people's risk perceptions and motivations to adopt preventive behavior is important in preventing the spread of recreational water illnesses (RWI) and other emerging infectious diseases. We developed a comprehensive scale measuring parents' perceived risk of their children contracting RWI. Parents (N = 263) completed a self-administered questionnaire with scale items based on 4 constructs of the Protection Motivation Theory: perceived vulnerability, perceived severity, response efficacy, and self-efficacy. Exploratory factor analysis identified 7 underlying factors, indicating 7 subscales of perceived risk for RWI. Cronbach alpha ranged from 0.60 to 0.81. The Precaution Adoption Process Model supported scale construct validity. This study provides the first perceived risk scale for exploring psychosocial factors that may predict or mediate the adoption of behaviors that prevent the spread of infectious diseases contracted by children while swimming. Findings from this study also provide implications for encouraging preventive behavior against other emerging infectious diseases. PMID- 15890118 TI - Avian influenza risk perception, Hong Kong. AB - A telephone survey of 986 Hong Kong households determined exposure and risk perception of avian influenza from live chicken sales. Householders bought 38,370,000 live chickens; 11% touched them when buying, generating 4,220,000 exposures annually; 36% (95% confidence interval [CI] 33%-39%) perceived this as risky, 9% (7%-11%) estimated >50% likelihood of resultant sickness, whereas 46% (43%-49%) said friends worried about such sickness. Recent China travel (adjusted odds ratio 0.35; CI 0.13-0.91), traditional beliefs (1.20, 1.06-1.13), willingness to change (0.29, 0.11-0.81) and believing cooking protects against avian influenza (8.66, 1.61-46.68) predicted buying. Birth in China (2.79, 1.43 5.44) or overseas (4.23, 1.43-12.53) and unemployment (3.87, 1.24-12.07) predicted touching. Age, avian influenza contagion worries, husbandry threat, avian influenza threat, and avian influenza anxiety predicted perceived sickness risk. High population exposures to live chickens and low perceived risk are potentially important health threats in avian influenza. PMID- 15890119 TI - Low diversity of Alkhurma hemorrhagic fever virus, Saudi Arabia, 1994-1999. AB - Alkhurma hemorrhagic fever virus (genus Flavivirus, AHFV) was recently identified as the agent of a viral hemorrhagic fever in Saudi Arabia and characterized serologically and genetically as a variant genotype of Kyasanur Forest disease virus (KFDV). Since viral diagnosis and vaccine development may be hindered by genetic diversity, this study was intended to address AHFV genetic heterogeneity. Eleven strains isolated from hospitalized patients from 1994 to 1999 in Saudi Arabia were sequenced in the envelope, NS3, and NS5 genes. Homologous sequences were compared and used to look for patterns reflecting specific evolution associated with spatiality, temporality, infection pathway, and disease prognosis. Genetic analyses showed low diversity, which suggests a slow microevolution. Evaluation of divergence times showed that AHFV and KFDV ancestral lineage diverged 66-177 years ago, and the diversity observed within the studied AHFV strains reflected a 4- to 72-year period of evolution. PMID- 15890120 TI - Osler and the infected letter. AB - The spread of infectious agents through the mail has concerned public health officials for 5 centuries. The dissemination of anthrax spores in the US mail in 2001 was a recent example. In 1901, two medical journals reported outbreaks of smallpox presumably introduced by letters contaminated with variola viruses. The stability and infectivity of the smallpox virus are reviewed from both a historical (anecdotal) perspective and modern virologic studies. Bubonic plague was the contagious disease that led to quarantines as early as the 14th century in port cities in southern Europe. Later, smallpox, cholera, typhus, and yellow fever were recognized as also warranting quarantine measures. Initially, attempts were made to decontaminate all goods imported from pestilential areas, particularly mail. Disinfection of mail was largely abandoned in the early 20th century with newer knowledge about the spread and stability of these 5 infectious agents. PMID- 15890121 TI - Salmonella Derby clonal spread from pork. AB - The genetic diversity of the Derby serotype of Salmonella enterica in Spain was examined by pulsed-field gel electrophoresis (PFGE). Out of 24 identified PFGE profiles, a major clone was detected in 19% of strains from humans, 52% from food, and 62% from swine. This clone (clone 1) was isolated from pork products, suggesting swine as its source. PMID- 15890122 TI - Probable tiger-to-tiger transmission of avian influenza H5N1. AB - During the second outbreak of avian influenza H5N1 in Thailand, probable horizontal transmission among tigers was demonstrated in the tiger zoo. Sequencing and phylogenetic analysis of those viruses showed no differences from the first isolate obtained in January 2004. This finding has implications for influenza virus epidemiology and pathogenicity in mammals. PMID- 15890123 TI - Highly pathogenic H5N1 influenza virus in smuggled Thai eagles, Belgium. AB - We report the isolation and characterization of a highly pathogenic avian influenza A/H5N1 virus from Crested Hawk-Eagles smuggled into Europe by air travel. A screening performed in human and avian contacts indicated no dissemination occurred. Illegal movements of birds are a major threat for the introduction of highly pathogenic avian influenza. PMID- 15890124 TI - Human T-cell leukemia virus type 1 molecular variants, Vanuatu, Melanesia. AB - Four of 391 Ni-Vanuatu women were infected with variants of human T-cell leukemia virus type 1 (HTLV-1) Melanesian subtype C. These strains had env nucleotide sequences approximately 99% similar to each other and diverging from the main molecular subtypes of HTLV-1 by 6% to 9%. These strains were likely introduced during ancient human population movements in Melanesia. PMID- 15890125 TI - Clonal comparison of Staphylococcus aureus isolates from healthy pig farmers, human controls, and pigs. AB - Pig farming is a risk factor for increased nasal Staphylococcus aureus colonization. Using sequence typing and phylogenetic comparisons, we showed that overcolonization of farmers was caused by a few bacterial strains that were not present in nonfarmers but often caused swine infections. This finding suggests a high rate of strain exchange between pigs and farmers. PMID- 15890126 TI - Human pythiosis, Brazil. AB - Pythiosis, caused by Pythium insidiosum, occurs in humans and animals and is acquired from aquatic environments that harbor the emerging pathogen. Diagnosis is difficult because clinical and histopathologic features are not pathognomonic. We report the first human case of pythiosis from Brazil, diagnosed by using culture and rDNA sequencing. PMID- 15890127 TI - Travel-associated rabies in Austrian man. AB - Rabies developed in an Austrian man after he was bitten by a dog in Agadir, Morocco. Diagnosis was confirmed by reverse transcription-polymerase chain reaction and immunohistochemistry. The patient's girlfriend was bitten by the same dog, but she did not become ill. PMID- 15890128 TI - Antibody testing and Lyme disease risk. AB - Lyme disease test results for >9,000 dogs were collected from participating veterinary clinics. Testing was conducted by using the IDEXX 3Dx kit, used widely by Maine veterinarians to screen clinically normal dogs during heartworm season. This study demonstrates how this test can be a valuable public health disease surveillance tool. PMID- 15890129 TI - Incident tuberculosis among recent US immigrants and exogenous reinfection. AB - Mathematical models and molecular epidemiologic investigation support the argument that exogenous reinfection plays an important role in tuberculosis transmission in high-incidence regions. We offer additional data from tuberculosis cases among recent US immigrants which strengthen the claim that reinfection in areas of intense transmission is common. PMID- 15890130 TI - Emergency survey methods in acute cryptosporidiosis outbreak. AB - In August 2003, a communitywide outbreak of cryptosporidiosis occurred in Kansas. We conducted a case-control study to assess risk factors associated with Cryptosporidium infection by using the telephone survey infrastructure of the Behavioral Risk Factor Surveillance System. Using existing state-based infrastructure provides an innovative means for investigating acute outbreaks. PMID- 15890131 TI - Media effects on students during SARS outbreak. AB - A few months after the 2003 severe acute respiratory syndrome (SARS) outbreak, a sample of Canadian undergraduate university students completed a questionnaire that showed that, despite believing media coverage of the outbreak was excessive, they had little anxiety about acquiring SARS. Additionally, 69% of participants failed a SARS-specific knowledge section of the questionnaire. PMID- 15890132 TI - Dengue fever, Hawaii, 2001-2002. AB - Autochthonous dengue infections were last reported in Hawaii in 1944. In September 2001, the Hawaii Department of Health was notified of an unusual febrile illness in a resident with no travel history; dengue fever was confirmed. During the investigation, 1,644 persons with locally acquired denguelike illness were evaluated, and 122 (7%) laboratory-positive dengue infections were identified; dengue virus serotype 1 was isolated from 15 patients. No cases of dengue hemorrhagic fever or shock syndrome were reported. In 3 instances autochthonous infections were linked to a person who reported denguelike illness after travel to French Polynesia. Phylogenetic analyses showed the Hawaiian isolates were closely associated with contemporaneous isolates from Tahiti. Aedes albopictus was present in all communities surveyed on Oahu, Maui, Molokai, and Kauai; no Ae. aegypti were found. This outbreak underscores the importance of maintaining surveillance and control of potential disease vectors even in the absence of an imminent disease threat. PMID- 15890133 TI - Dengue risk among visitors to Hawaii during an outbreak. AB - Despite the high rates of dengue in many tropical destinations frequented by tourists, limited information is available on the risk for infection among short term visitors. We retrospectively surveyed 4,000 persons who arrived in Hawaii during the peak of the 2001-2002 dengue outbreak and collected follow-up serologic test results for those reporting denguelike illness. Of 3,064 visitors who responded, 94 (3%) experienced a denguelike illness either during their trip or within 14 days of departure; 34 of these persons were seen by a physician, and 2 were hospitalized. Twenty-seven visitors with denguelike illness provided a serum specimen; all specimens were negative for anti-dengue immunoglobulin G antibodies. The point estimate of dengue incidence was zero infections per 358 person-days of exposure with an upper 95% confidence limit of 3.0 cases per person-year. Thus, the risk for dengue infection for visitors to Hawaii during the outbreak was low. PMID- 15890134 TI - Dengue type 3 virus, Saint Martin, 2003-2004. AB - We describe the spread of a dengue virus during an outbreak in Saint Martin island (French West Indies) during winter 2003-2004. Dengue type 3 viruses were isolated from 6 patients exhibiting clinical symptoms. This serotype had not been detected on the island during the preceding 3 years. Genome sequence determinations and analyses showed a common origin with dengue type 3 viruses isolated in Martinique 2 years earlier. PMID- 15890135 TI - Dengue antibody prevalence in German travelers. AB - We studied 2,259 German citizens after they returned from dengue-endemic countries from 1996 to 2004. Serotype-specific dengue antibodies indicated acute infections in 51 (4.7%) travelers with recent fever and 13 (1.1%) travelers with no recent fever, depending largely on destination and epidemic activity in the countries visited. PMID- 15890136 TI - Leptospirosis during dengue outbreak, Bangladesh. AB - We collected acute-phase serum samples from febrile patients at 2 major hospitals in Dhaka, Bangladesh, during an outbreak of dengue fever in 2001. A total of 18% of dengue-negative patients tested positive for leptospirosis. The case-fatality rate among leptospirosis patients (5%) was higher than among dengue fever patients (1.2%). PMID- 15890137 TI - Retinal hemorrhages in 4 patients with dengue fever. AB - We report 4 patients with retinal hemorrhages that developed during hospitalization for dengue fever. Onset of symptoms coincided with resolution of fever and the nadir of thrombocytopenia. Retinal hemorrhages may reflect the rising incidence of dengue in Singapore or may be caused by changes in the predominant serotype of the dengue virus. PMID- 15890138 TI - A randomised controlled comparison of alternative strategies in stroke care. AB - OBJECTIVES: To compare outcomes between stroke patients managed on the stroke unit, on general wards with stroke team support or at home by specialist domiciliary team and to derive prognostic variables that will identify patients most suitable for management by each strategy. To describe organisational aspects of individual strategies of stroke care and to evaluate cost-effectiveness of each strategy and its acceptability to patients, carers and professionals. DESIGN: Prospective single-blind randomised controlled trial undertaken in patients recruited from a community-based stroke register. SETTING: Suburban district in south-east England. PARTICIPANTS: Patients with disabling stroke who could be supported at home. INTERVENTIONS: The stroke unit gave 24-hour care provided by specialist multidisciplinary team based on clear guidelines for acute care, prevention of complications, rehabilitation and secondary prevention. The stroke team provided management on general wards with specialist team support. The team undertook stroke assessments and advised ward-based nursing and therapy staff on acute care, secondary prevention and rehabilitation aspects. Domiciliary care involved management at home under the supervision of a GP and stroke specialist with support from specialist team and community services. Support was provided for a maximum of 3 months. MAIN OUTCOME MEASURES: The primary measure was death or institutionalisation at 1 year. Secondary measures were dependence, functional abilities, mood, quality of life, resource use, length of hospital stay, and patient, carer and professional satisfaction. RESULTS: Of the 457 patients randomised, 152 patients were allocated to the stroke unit; 152 patients to stroke team and 153 patients to domiciliary stroke care (average age 76 years, 48% women). The groups were well matched for baseline characteristics, stroke type and severity, level of impairment and initial disability. Fifty-one (34%) patients in the domiciliary group were admitted to hospital after randomisation. Mortality and institutionalisation at 1 year were lower on stroke unit compared with stroke team or domiciliary care. Significantly fewer patients on the stroke unit died compared with those managed by the stroke team. The proportion of patients alive without severe disability at 1 year was also significantly higher on the stroke unit compared with stroke team or domiciliary care. These differences were present at 3 and 6 months after stroke. Stroke survivors managed on the stroke unit showed greater improvement on basic activities of daily living compared with other strategies. Achievement of higher levels of function was not influenced by strategy of care. Quality of life at 3 months was significantly better in stroke unit and domiciliary care patients. There was greater dissatisfaction with care on general wards compared with stroke unit or domiciliary care. Poor outcome with domiciliary care and on general wards was associated with Barthel Index <5, incontinence and, on general wards, age over 75 years. The total costs of stroke per patient over the 12-month period were pound 11,450 for stroke unit, pound 9527 for stroke team and pound 6840 for home care. However, the mean costs per day alive for the stroke unit were significantly less than those for the specialist stroke team patients, but no different from domiciliary care patients. Costs for the domiciliary group were significantly less than for those managed by the specialist stroke team on general wards. CONCLUSIONS: Stroke units were found to be more effective than a specialist stroke team or specialist domiciliary care in reducing mortality, institutionalisation and dependence after stroke. A role for specialist domiciliary services for acute stroke is not supported by this study. Management of patients with strokes on general medical wards, even with specialist team input, is not supported by this study. The stroke unit intervention was less costly per patient day alive and more effective than the stroke team intervention. The stroke unit was more effective and of equivalent cost when compared to home care. Hence, the stroke unit is a more cost-effective intervention than either stroke team or home care. Further research is needed to understand processes contributing to the reduction in mortality on stroke units and to determine the generalisability of these results and the factors that will influence the implementation of the findings of this study in clinical practice. PMID- 15890139 TI - The investigation and analysis of critical incidents and adverse events in healthcare. AB - OBJECTIVES: To carry out a review of published and unpublished work on the analysis on methods of accident investigation in high-risk industries, and of critical incidents in healthcare. To develop and pilot guidelines for the analysis of critical incidents in healthcare for the hospital sector, mental health and primary care. DATA SOURCES: Literature already available in the Clinical Risk Unit, University College London. Work by known experts in the field of accident investigation and analysis. Electronic databases including PsycINFO and MEDLINE. Websites for accident investigation reports. REVIEW METHODS: Twelve techniques from other high-risk industries were reviewed in detail using criteria developed for the purpose. This review provided a conceptual framework for the healthcare review and appraisal process, as well as providing a critical assessment of the industry techniques. Rigorous searching and screening identified 138 papers for formal appraisal and a further 114 were designated as providing potentially useful background information. A formal appraisal instrument was designed, piloted and modified until acceptable reliability was achieved. From the 138 papers, six techniques were identified as representing clearly definable approaches to incident investigation and analysis. All relevant papers were reviewed for each of the six techniques: Australian Incident Monitoring System, the Critical Incident Technique, Significant Event Auditing, Root Cause Analysis, Organisational Accident Causation Model and Comparison with Standards approach. RESULTS: All healthcare techniques had the potential of being applied in any specialty or discipline related to healthcare. While a few studies looked solely at death as an outcome, most used a variety of outcomes including near misses. Most techniques used interviewing and primary document review to investigate incidents. All techniques included papers that identified clinical issues and some attempt to assess underlying errors, causes and contributory factors. However the extent and sophistication of the various attempts varied widely. Only a third of papers referred to an established model of accident causation. In most studies examined there was little or no information on the training of investigators, how the data was extracted or any information on quality assurance for data collection and analysis. There was some variation in the level of expertise and training required but to undertake the investigation to an acceptable depth all required some expertise. In most papers there was little or no discussion of implementation of any changes as a result of the investigations. A quarter of publications gave some description of the implementation of changes, though few addressed evaluation of changes. CONCLUSIONS: The reviews demonstrate that, while much valuable work has been accomplished, there is considerable potential for further development of techniques, the utilisation of a wider range of techniques and a need for validation and evaluation of existing methods which would make incident investigation more versatile and use limited resources more effectively. Further exploration of techniques used in high-risk industries, with interviews and observation of actual investigations should prove valuable. Existing healthcare techniques would benefit from formal evaluation of their outcomes and effectiveness. Studies should examine depth of investigation and analysis, adequacy and feasibility of recommendations and cost effectiveness. Examining implementation of recommendations is a key issue. PMID- 15890140 TI - Aspects of internal fixation of fractures in porotic bone. Principles, technologies and procedures using locked plate screws. AB - Fractures of the bones of elderly people occur more often and have a more important effect because of a generally diminished ability to coordinate stance and walking. These fractures occur at a lower level of load because of lack of strength of the porotic bone. Prompt recovery of skeletal support function is essential to avoid respiratory and circulatory complications in the elderly. To prevent elderly people from the risks of being bedridden, demanding internal fixation of fractures is required. The weak porotic bone and the high level of uncontrolled loading after internal fixation pose complex problems. A combination of several technical elements of design, application and aftercare in internal fixation are proposed. Internal fixators with locked screws improve the biology and the mechanics of internal fixation. When such fixators are used as elevated splints they may stimulate early callus formation because of their flexibility, the limit of flexibility being set by the demands of resistance and function of the limb. Our own studies of triangulation of locked screws have demonstrated their beneficial effects and unexpected limitations. PMID- 15890141 TI - [Long-term results following fracture of the femoral neck in children]. AB - PURPOSE OF THE STUDY: This retrospective study was designed to evaluate the severity and nature of long-term sequelae of femoral neck fractures in children in relation to the strategy and technique of therapy. MATERIAL: The study included 15 patients with a fracture of the femoral neck who, at the time of injury, had an opened proximal physis. The average age at the time of injury was 11.5 years (range, 4 to 16.3 years). There were eight boys and seven girls. Twelve children suffered injury due to a fall varying in gravity, two were knocked down by a vehicle, and one was injured as a co-driver in a car accident. The group involved no type I fracture, six type II, seven type III and two type IV fractures, as classified by the Delbet and Colonna system. METHODS: All children were operated on within an average of 4.1 days after injury. The delay was caused by a late referral from an outside hospital or was due to associated complications.However, the majority of fractures were treated within 24 hours of injury. Surgery was carried out from the anterolateral approach. Miniarthrotomy was performed in 11 patients to remove hematoma and decompress the intra articular space. The methods of stabilization included Kirschner's wires in four children, compressive osteosynthesis using lag screws inserted extraphyseally in 10 children and the combination of both methods in one child. No plaster of Paris spica or traction was applied after surgery. RESULTS: The long-term results were evaluated at a minimum of 5 years after injury. The average follow-up was 9 years and 11 months, with the range from 5 years and 1 month to 15 years and 5 months, and 12 patients were involved. Six had a type II and six had a type III fracture. Ten patients were treated by arthrotomy. Stabilization of the fracture was performed with lag screws in nine children and Kirschner's wires in three patients. Both subjective and objective findings were evaluated by the modified Rattlif criteria. Concerning pain, eight (67 %) children had excellent outcomes, three (25 %) reported good outcomes and only one (8 %) had a poor outcome. The activity following injury was subjectively evaluated as excellent by 11 (92 %) patients and as good by one (8 %) patient. None of the patients was noticeably limited in their activity, as compared with the pre-injury state. Objective findings were based on X-ray images and the range of hip motion. No or minimal radiographic changes were found in five (42 %) patients and were assessed as excellent outcomes. A good outcome, i. e., a spherical head with a moderate neck deformity, was achieved in five (42 %) patients. A poor outcome, i. e., avascular necrosis free of revitalization, but with collapse, on X-ray images was recorded in two (16 %) patients. The values for the range of motion and limb-length discrepancy were excellent in eight (67 %), good in two (16.5 %) and poor in two (16.5 %) patients. Poor objective and subjective findings were recorded in the patients who had not undergone miniarthrotomy. DISCUSSION: A comparison with the literature data showed that the occurrence of each fracture type was in agreement with the reports of other authors. The finding of a higher frequency of type I fractures can be explained by a pathological slip due to hormonal changes. The opinions on treatment of these fractures have developed to the view that surgery is necessary in the early post-injury period, preferably with the use of compressive osteosynthesis or Kirschner's wires. However, subsequent immobilization in a plaster cast spica is not necessary. The subjective and objective findings were not exactly correlated due to differences in patients' age and different intervals between injury and treatment. CONCLUSIONS: The authors recommend early surgery and stabilization by compressive osteosynthesis or Kirschner's wires, together with miniarthrotomy in order to decompress the articular space. PMID- 15890142 TI - [Combined atlantoaxial fractures]. AB - PURPOSE OF THE STUDY: Combined fractures of the atlas and epistropheus account for 3 % of all acute injuries to the cervical spine. In relation to all C1 and C2 injuries this is 43 % and 16 %, respectively. The aim of this study is to evaluate a group of patients with combined C1-C2 fractures and to suggest an effective therapeutic procedure. MATERIAL: In the years 1996 to 2003, a total of 16 patients with trauma to the atlantoaxial complex were treated at the Orthopedic Department of the Third Faculty of Medicine, Charles University, Prague (1996-2001) and the Department of Spinal Surgery of the University Hospital in Motol, Prague (2001-2003). These injuries included a combined fracture of the dens (Anderson and D'Alonzo type II) and of the atlas posterior arch in six patients, a type II dens fracture combined with Jefferson fracture in two patients, a type III fracture of the dens with a lateral mass fracture in two patients, hangman's fracture with posterior arch fracture in three patients, a type II fracture of the dens with anterior arch fracture in one patient, a fracture of the C2 body with Jefferson fracture in one patient and a fracture of the C2 body with fracture of the lateral mass in one patient. Two patients were treated conservatively and 14 underwent surgery. On admission neurological deficit was found in five patients. METHODS: Fourteen patients were operated on. Direct osteosynthesis of the dens, with motion in the atlantoaxial complex preserved, was performed in five patients. Seven patients underwent C1-C2 fixation that, in one, involved the C1-C3 segments; five patients were treated by Harms fixation with polyaxial screws from the posterior approach, two by the Magerl or Gallie techniques and one patient required occipito-cervical fixation of C0-C2. The patient with a hangman's fracture combined with fracture of the atlas posterior arch was treated by discectomy of C2-C3, tricortical graft from the iliac crest and plate application. The patients used Philadelphia collars for 6 to 12 weeks according to the type of injury and their bone quality. RESULTS: Three patients (two undergoing direct osteosynthesis of the dens and one with occipito-cervical fixation) reported intermittent upper neck pain that required taking analgesics. The patient treated by occipito-cervical fixation repeatedly complained of restriction of rotational head movement by about 50 %. Radiograms of the cervical spine in both flexion and extension taken at 12- to 14-week follow-up all showed stable C0-C1 and C1-C2 segments. In the five patients undergoing direct osteosynthesis of the dens, complete bony union was found on X ray and CT examination by 6 to 24 weeks postoperatively. Similarly, full instrumented fusion was achieved by 12 to 24 weeks postoperatively in the seven patients treated by dorsal fixation. The patient with anterior C2-C3 fixation showed, on X-ray images, a completely remodeled segment at 24 weeks after surgery. There was one intraoperative complication involving management of profuse bleeding from the venous plexus along the greater occipital nerve. No other complications related to the surgical procedure were recorded and no injury to the spinal cord, nerve roots or the vertebral artery was observed. None of the patients experienced any deterioration of neurological findings during the early postoperative period. One patient had to undergo resuturing of the operative wound from the posterior approach, because of subcutaneous necrosis that had failed to heal. No instrumentation failure or infection, regarded as late complications, were recorded. DISCUSSION: At our Department we prefer early operative treatment involving spondylodesis in the shortest segment possible, with special emphasis on preserving rotational C1-C2 movement. Therefore, in some cases, we use only temporary stabilization with removal of instrumentation after 3 to 4 months. In this group the most frequent fractures were those combined with type II fractures of the dens. In such cases we always prefer direct osteosynthesis of the dens or, if this is not possible, the Harms technique of C1 C2 fixation, possibly only temporary. We believe, in agreement with Guilot and Fesser, that a potential failure of conservative therapy may result in a longer convalescent period and that patients should always be informed about these issues. In contrast to Guilot and Fesser we treat combined hangman's fractures from the anterior approach, by discectomy, tricortical graft and plate application. CONCLUSIONS: Combined atlantoaxial fractures are serious, life threatening injuries which, because of their diversity, require an individual approach to each patient. Early surgery is recommended with increasing frequency, particularly in the cases with persisting dislocation or instability. At the same time it is necessary to ensure that motion restriction of the cervical spine be minimal. PMID- 15890143 TI - [Arthrodesis of interphalangeal joints by means of external frame fixation]. AB - PURPOSE OF THE STUDY: The indications for arthrodesis of interphalangeal joints include pain, instability, deformity or irreparable damage to the relevant flexor or extensor tendon. The general principle of arthrodesis is to resect the affected joint ends in the flexion desired so that a highest possible surface of contact may be provided. Subsequently, retention by means of internal or external fixation is performed. The aim of this study is to evaluate our method of interphalangeal joint arthrodesis, using an external frame fixator, to present the results and to point out the advantages of this method, thus showing its applicability for relevant surgical indication. MATERIAL: Arthrodesis by external fixation was carried out in 37 patients. The group included 21 men and 16 women at an average age of 49.6 years (range, 30 to 67 years). The method was used in the patients who differed in the etiology of lesions of interphalangeal joints. Twenty-two patients had previously experienced purulent arthritis, six had post traumatic arthritis and joint instability, five had rheumatoid arthritis, two had primary arthritis and two had an inveterate rupture of the extensor aponeurosis. METHODS: We used a simple frame fixator whose stability was based on two to four Kirschner's wires inserted and maintained in clamps on rods, 4 cm or 7.5 cm long, each having a double anti-clockwise thread that facilitates compression, distraction and correction in one plane. The surgical procedure was carried out under axillary block with the use of a tourniquet. It involved making an S-shaped incision dorsally, severing the extensor apparatus, resecting articular surfaces in the desired flexion position, inserting parallel wires and fixing them in clamps on rods, achieving compression, checking the position of articular surfaces and suturing. Included in the group assessment were the gender and age of the patients, etiology of articular disease, number of the digits and joints affected, limb laterality, wound healing, stability of fixation, maintenance of a correct arthrodesis position and signs of healing on X-ray images. Time required for bony union and the shortening of the digit due to surgery were also included in the evaluation. RESULTS: The 35 followed-up patients showed healing of the wound and arthrodesis, with firm bony union being achieved at an average of 6.7 weeks. In one patient after removal of the fixator, septic pseudoarthrosis developed at the site of the resected distal interphalangeal joint. Another patient developed ischemia of the distal phalanx of the thumb treated, which required release and subsequent removal of the external fixator, with vasodilatation therapy. After a prolonged topical therapy, painless fibrous ankylosis developed in that interphalangeal joint and the thumb was salvaged. DISCUSSION: The aim of arthrodesis is to achieve firm and painless bony union in a correct functional position at a reasonable time. Arthrodesis in our patients healed at an average time of 6.7 weeks. The high effectivity of the compression technique in achieving firm bony fusion is the factor emphasized in the literature, particularly in situations where there is a reduced contact surface, poor coverage by soft tissue, infection or the presence of a systemic disease, such as rheumatoid arthritis or diabetes mellitus. This all is in agreement with the results of this study. CONCLUSIONS: This study shows advantages of the compression technique of arthrodesis by means of external frame fixation based on insertion of wires beyond the site of inflammation. Compression and stability result in rapid osseous union, immobilization in plaster cast is not necessary, free joints of the hand can be exercised, the fixator is removed in an outpatient department and the minimal presence of metal material does not interfere with good healing of soft tissues. Therefore this method has all merits to be used for surgery in a terrain affected by rheumatic, inflammatory or potentially inflammatory lesions. PMID- 15890144 TI - Tribological considerations for a new hip system. AB - PURPOSE OF THE STUDY: This paper was initiated to evaluate a new total hip system based on cementless fixation and focusing options for different articulations within a single hip system. MATERIAL AND METHODS: 100 patients provided with the cementless Variall hip system were evaluated clinically and radiographically. The 59 female and 41 male patients were followed for a minimum of 3 years postoperative. They were clinically evaluated using the Harris Hip Score. Radiographic analysis was based on plain x-rays at the latest follow-up evaluation. RESULTS: The evaluated patients achieved an excellent clinical result with an average Harris Hip Score of 94,3 points after an average follow-up of 42 months. The radiographic evaluation of the threaded cup did not exhibit any migration of the implant nor could we find radiolucencies. All tapered stems were radiographically stable. Due to the modified proximal design and the macrostructure an improved stem bone interface was found during the follow-up period. DISCUSSION The new cementless cup with its cylindrical threads and the spherical floor achieves an anatomic fit within the acetabulum and provides excellent primary stability. The design of the gamma inlay offers the fixation of ceramic and metal articulations without polyethylene interface. Additionally, options for conventional and cross-linked polyethylenes are available. These facts make the cup a cost-effective device. The design of the new uncemented stem guarantees a high degree of primary stability and excellent rotational stability due its rectangular cross section. This pressfit is further enhanced by the proximal macrostructure of the implant, again ensuring an excellent bond between prosthesis and bone. CONCLUSION: Thanks to the versatility of all the components this comprehensive new total hip prosthesis offers many options for a long-term successful implant. Additionally, it is a cost-effective solution - an important fact of nowadays strained financial situation in public health care. PMID- 15890145 TI - [Forefoot surgery under regional anesthesia]. AB - PURPOSE OF THE STUDY: The aim of this study is to present the method of lower limb peripheral nerve block for forefoot surgery and, in comparison with other methods, to evaluate the results in terms of postoperative analgesia and postoperative complications. MATERIAL: Our group included 164 patients, 13 men and 151 women in the age range from 16 to 79 years, who underwent surgery for forefoot deformities during the years 1998-99. Of these, 77 (46 %) were operated on under general anesthesia, 54 (33 %) under infiltration anesthesia and 33 (21 %) under peripheral nerve block. METHODS: When peripheral nerve block was used, the anesthetic Marcain was administered about 1 hour before surgery at the following sites: along the deep branch of the peroneal nerve on the dorsal side of the foot, along the tibial nerve behind the inner ankle, and along the sural nerve in the outer ankle region. After surgery, the duration of requirement for analgesia in all three types of anesthesia was recorded. Complications associated with wound healing were also recorded. Subjective satisfaction of the patients was evaluated as a separate issue. RESULTS: No intraoperative complications were found in the patients operated on under peripheral nerve block. In three patients, regional anesthesia was not sufficient and had to be completed with topical anesthetics administered during the operation. The patients treated under regional anesthesia asked for analgesics after an average of 9.5 hours; those operated on under infiltration and general anesthesia required analgesics at an average of 2.5 hours (range, 2 to 4 hours) and within 30 min of waking up, respectively. Complications associated with wound healing were recorded in 5 %, 11 % and 6 % of the patients treated under general, infiltration and regional anesthesia, respectively. Of the 33 patients operated on under regional anesthesia, 28 would prefer this anesthesia for any other similar surgery and five would rather have general anesthesia. DISCUSSION: Regional anesthesia is associated with fewer complications of wound healing than topical anesthesia, which is apparently due to a lower degree of ischemia produced in the treated limb by peripheral nerve block. Healing after regional anesthesia is only a little worse than after general anesthesia, but with the absence of risks related to general anesthesia. Patients operated on under general anesthesia feel pain immediately after surgery, those receiving infiltration anesthesia ask for painkillers at an average of 2.5 hours and, with well-introduced regional anesthesia, patients are free from pain for 9.5 hour on the average. The duration of postoperative pain absence is also related to the anesthetic used. The use of 1 % Mesocain results in a fast onset of anesthesia but its effect is shorter, while 0.5 % Marcain has a slower onset but a longer effect. CONCLUSIONS: Regional anesthesia for forefoot surgery, if the anesthetic is well administered, is one of the options associated with minimal risks. It apparently reduces complications of wound healing, when compared with infiltration anesthesia. It can be used in situations where general anesthesia would put the patient at risk. It also has a pronounced analgesic effect that persists well after surgery and provides better postoperative comfort for the patient. Key words: lower limb, peripheral nerve block, forefoot surgery. PMID- 15890146 TI - [Unusual infection complication of total hip arthroplasty]. AB - Authors present the case history of a 66-year old patient after repeated reimplantations of the THA with a deep infect caused by a rare aetiological agent (Serratia marcescens) associated with a pyogenic sinus. They describe the disease history, therapeutic procedure, complications associated with the surgery as well as postoperative course after the reimplantation of a customized total hip replacement. In the conclusion they state that in case of an infected total hip arthroplasty the treatment is focused on the salvage of the infection process and preservation of the function of the affected limb. Of essential importance is surgical revision with a radical removal of necrotic tissues and hardware in combination with an intensive parenteral antibiotic administration. PMID- 15890147 TI - [Primary surgical treatment of amputated fingers and indications for digital replantation]. AB - The amputation of a single finger, or its part, or more fingers results in functional and esthetic changes in the patient's life. Until 1965 when the first thumb ever was replanted, the treatment of amputated digits had been limited by technical facilities of the medical science. Since 1970s, the development of fine suture materials, microsurgical instruments and the operating microscope has made it possible that replantations have become routine procedures in hand surgery. Both surgical procedures and indication schemes have also evolved. The primary surgical treatment has been standardized to involve the wrapping of amputated parts in dressing material saturated with isotonic solution and cooling at 4 to 10 degrees C during transport. The first enthusiasm for replantation of everything that had been amputated was replaced, owing to long-term post operative results, by a more selective approach. Even an absolute indication for digital replantation, such as amputation of a thumb, two or more fingers, amputation in the palm and all amputations in children, must be put aside when life-threatening injuries or serious diseases are present. The benefit of replantation should always outweigh the trauma of any operative procedure because this must not harm the patient. PMID- 15890148 TI - Prognostic factors in neuroblastoma: the need for evidence-based decisions in prospective studies. PMID- 15890149 TI - A panel discussion of controversies and challenges in the adjuvant treatment of colon cancer. AB - Current issues of adjuvant therapy for colon cancer concern the introduction of drugs other than fluorouracil-5/leucovorin (5-FU/LV), the benefits for stage II patients, the use of new primary endpoints and the influence of age on treatment benefits. These issues were addressed in a panel discussion and the conclusions were the following: FOLFOX4 is the first regimen that shows superiority over 5 FU/LV. The use of 3-year disease-free survival as primary endpoint could encourage the quicker adoption of improved therapeutic strategies into clinical practice. Available data suggest that there are some benefits for stage II patients, and the decision needs to be individualised for each patient. Further, therapeutic decisions based solely on the patient's age are inappropriate, and geriatric assessment tools will help in making this decision. This information would improve patient and physician understanding of the recent data regarding the potential benefits of adjuvant therapy. PMID- 15890150 TI - Prognostic factors including neoadjuvant chemotherapy in Mexican children with neuroblastoma. AB - INTRODUCTION: There are several prognostic factors in children with neuroblastoma that have been outlined in the international literature. MATERIAL AND METHODS: A retrospective study was carried out analysing the medical records of patients with the pathological diagnosis of neuroblastoma seen at the Department of Oncology from the Instituto Nacional de Pediatria (Mexico) between January 1984 to January 1997. A total of 32 clinical prognostic factors were assess in our population. RESULTS: Fifty five patients whose age ranged from 1 to 168 months old, mean of 35 months were included. Out of 32 prognostic factors only 6 including sex (p= 0.0039), metastatic disease to bone (p= 0.003), bone marrow involvement (p= 0.0027), staging system (p= 0.000015), surgical treatment (p 0,0022) and neoadjuvant chemotherapy (p.005) were the most significant. CONCLUSIONS: It was concluded that besides the prognostic factors outlined, neoadjuvant chemotherapy is of utmost importance. It decreases tumor volume and allows surgery to be more successful, therefore believing that this variable represents a specific prognostic factor in cases of advanced neuroblastoma. PMID- 15890151 TI - Predictors of positive axillary lymph nodes in breast cancer patients with metastatic sentinel lymph node. AB - INTRODUCTION: Breast cancer with metastatic sentinel lymph nodes (SLN) may have clinico-pathologic factors associated with the presence of positive non-sentinel axillary nodes (NSLN). The aim of the present study was to determine factors that predict involvement of NSLN in breast cancer patients with positive SLN. MATERIAL AND METHODS: A prospective database search identified 80 patients who underwent SLN biopsy for invasive breast cancer between January 1999 and August 2002. Clinico-pathologic data was analyzed to determine factors that predicted additional positive axillary nodes. RESULTS: A total of 23 patients had positive SLN and underwent conventional axillary lymph node dissection. Statistical analysis revealed that lympho-vascular invasion (p~0.00000), SLN metastasis >2 mm (p=0.002), and the presence of extra-nodal involvement (p=0.002), were positive predictors of the metastatic involvement of NSLN. CONCLUSIONS: The likelihood of positive NSLN correlates with pathologic parameters such as the presence of lympho-vascular invasion, size of the SLN metastasis, and extra-nodal involvement. These data may be helpful with the regard to the decision to undertake axillary dissection in breast cancer patients with metastatic sentinel lymph nodes. PMID- 15890152 TI - Carboplatin and tegafur-uracil concomitant with standard radiotherapy in the management of locally advanced head and neck cancer. AB - INTRODUCTION: We undertook a prospective study to determine the feasibility, toxicity, response and survival rate of simultaneous chemotherapy (CT) and radiotherapy (RT) for locally-advanced head and neck cancer. MATERIAL AND METHODS: Fifty eight patients were treated with carboplatin (i.e. 100 mg/m(2)) weekly, tegafur-uracil (UFT) (oral 400 mg/m(2)) daily and simultaneous treatment with a cobalt-60 source of radiation (total dose 65-70 Gy). RESULTS: Forty six patients (79%) received the total dose of RT while CT was delayed or reduced in 31 patients (53%). Grade 3-4 toxicity observed was mucositis in 27 (47%), leukopenia in 10 (17%), anaemia in 5 (9%), and diarrhoea in 4 (7%) patients. The objective response rate was 74%; 24 complete response (41%) and 19 partial response (33%). Overall, there are 11 patients (19%) disease-free, 7 (12%) alive with disease, 35 have died of progressive disease (60%) and 3 (5%) from other causes. There were 2 toxic deaths (3%). Median time to progression was 10 months and median survival was 18.4 months. CONCLUSIONS: The use of carboplatin and UFT concomitant with radiotherapy has, in our study, a slightly lower activity than other chemo-radiotherapy protocols, especially with respect to complete responses, but with no significant differences in overall survival or disease free survival rates. PMID- 15890153 TI - Subacute dementia as presenting feature of carcinomatous leptomeningeal metastases. PMID- 15890154 TI - Chromoendoscopic mucosal resection of a gastric dysplasia. PMID- 15890155 TI - Paediatric oncology: embryonic tumours. PMID- 15890157 TI - [Open letter to the Sociedad Espanola de Oncologia Quirurgica]. PMID- 15890156 TI - Rhabdomyosarcoma: present and future perspectives in diagnosis and treatment. PMID- 15890158 TI - Structural Approaches to Sequence Evolution: Molecules, Networks and Populations. Proceedings of a workshop, 5-10 July 2004, Dresden, Germany. PMID- 15890159 TI - The repeatability of corticomotor threshold measurements. AB - Threshold (Th) is a neurophysiological parameter frequently used in TMS studies. The present study was designed to investigate the repeatability of the Th measurements by reexamining healthy subjects over various time points. Overall, 82 subjects (median age: 19 years, range: 12-65) entered the study. Following a baseline examination, there were six retest sessions: S0 (n = 8 hemispheres reexamined, mean interval x = 19 min), S1 (n = 34 hemispheres reexamined, mean interval x = 4 days), S2 (n = 32 hemispheres, x = 29 days), S3 (n = 30 hemispheres, x = 106 days), S4 (n = 30 hemispheres, x = 183 days) and S5 (n = 30 hemispheres, x = 1867 days). Stimulation was performed with a figure of eight coil and Th was defined at 1% steps. At baseline, controls had an MT of 41.1 +/- 8. Mean difference of MT from baseline was 0.62 on S0 (95% confidence interval (CI) of the difference: -1.04 to +2.29), 0.13 on S1 (95% CI: -1.2 to +1.5), -0.03 on S2 (95% CI: -1.1 to +1.06), -2.07 on S3 (95% CI: -4.33 to +0.19), 0.15 on S4 (95% CI: -0.98 to +1.28) and 0.87 on S5 (95% CI: -0.49 to +2.23). None of these differences were statistically significant (repeated measures ANOVA, P > 0.05). The upper limit of MT difference that an individual subject might have with a probability of 95% (measurement error) was 8. The repeatability of the method was found to be independent from the age of the subjects, the magnitude of threshold or the test-retest interval. The topography of corticomotor threshold was also investigated. Minimal threshold values were obtained from a restricted area of scalp sites that always included the fixed stimulation point of the current protocol. Therefore, using a fixed stimulation point is an adequate technique for measuring threshold. In conclusion, threshold is a stable parameter on an individual and group basis. These data quantify the repeatability of the method and may prove useful in the interpretation of findings during longitudinal studies. PMID- 15890160 TI - Paradigm design of sensory-motor and language tests in clinical fMRI. AB - Functional magnetic resonance imaging (fMRI) paradigms on sensory-motor and language functions are reviewed from a clinical user's perspective. The objective was to identify special requirements regarding the design of fMRI paradigms for clinical applications. A wide range of methods for setting up fMRI examinations were found in the literature. It was concluded that there is a need for standardised procedures adapted for clinical settings. Sensory-motor activation patterns do not vary much at different hand motion tasks. Nevertheless it is one of the most important clinical tests. In contrast, the language system is much more complex. In several studies it has been observed that word production tasks are preferable in determination of language lateralisation. Broca's area is activated by most tasks, whereas sentence processing and semantic decision also involve activation in temporoparietal and frontal areas. However, combined task analysis (CTA) of several different tasks has been found to be more robust and reliable for clinical fMRI compared to separate task analysis. PMID- 15890161 TI - [Methods and clinical value of peripheral nerve refractory period measurement in man]. AB - Immediately after action potential occurrence, owing to transient sodium channel inactivation, axon excitability is reduced for a short period of time, including the absolute refractory period, a first period of total inexcitability, followed by the relative refractory period. There are basically two different stimulation protocols to estimate axonal refractoriness, i.e. "paired-pulse" and "collision" techniques. Refractory period has been assessed in various conditions and appeared to depend on several physiological or methodological factors, featuring the type of nerve or the characteristics of the subject, but also the technique of stimulation or the method of data analysis. In addition, refractory periods can be altered by pathological conditions. Several studies showed prolonged refractory periods in patients suffering from alcoholic, diabetic or toxic neuropathies. Refractory period abnormality is a sensitive marker of axonal dysfunction as observed in Guillain-Barre syndrome, carpal tunnel syndrome or multiple sclerosis. Thus, the measurement of the refractory periods is a valuable tool to study the pathophysiology of peripheral nerves, complementary to standard nerve conduction studies. However, the application of these techniques in the routine practice of clinical neurophysiology remains limited. PMID- 15890162 TI - What is the clinical significance of periodic limb movements during sleep? AB - To understand better the clinical impact of periodic limb movements during sleep (PLMS) we analysed data from 51 patients who, following an adaptation night, presented a PLMS index > 5 during two consecutive nocturnal polysomnographic recordings. In the morning following each recording patients completed a questionnaire including five visual analogic scales (VAS): (1) I did not sleep well/I slept very well. (2) I feel very sleepy/I do not feel sleepy at all. (3) I feel very tired/I feel very dynamic. (4) Physically, I do not feel fit/physically, I feel fit. (5) Psychologically, I do not feel fit/psychologically, I feel fit. We compared the responses to these questions with the PLMS index, first inter-individually, then intra-individually between nights. RESULTS: The inter-individual analysis did not show correlations between the PLMS index and the questions (1) and (2). We found a significant correlation between the PLMS index and the questions (3) (r = -0.29; P < 0.05), (4) (r = 0.30; P < 0.05) and (5) (r = -0.39; P < 0.01). For the intra-individual analysis, we did not find correlations between the PLMS index and questions (1)-(3), but found a significant correlation with questions (4) (r = -0.28; P < 0.05) and (5) (r = -0.36; P < 0.01). CONCLUSION: PLMS per se, or the sleep changes induced by them, seem to be associated with decreased physical and psychological fitness on awakening. PMID- 15890163 TI - Eating disorder NOS (EDNOS): an example of the troublesome "not otherwise specified" (NOS) category in DSM-IV. AB - The "Not Otherwise Specified" (NOS) category within DSM-IV is designed for disorders of clinical severity that are not specified within broad diagnostic classes. "NOS" diagnoses are intended to be residual categories and they tend to be neglected by researchers. This can be inappropriate. The problems associated with certain NOS diagnoses are well illustrated by "Eating Disorder NOS" (sometimes termed EDNOS), which is the most common category of eating disorder encountered in routine clinical practice yet it has barely been studied. Indeed, there has been no research on its treatment. Interim and longer-term conceptual and practical solutions to the anomalous status of eating disorder NOS are proposed including the creation of a new diagnosis termed "mixed eating disorder". Several of these solutions are of relevance to NOS categories in general. All the solutions should fulfil criteria for clinical utility. PMID- 15890164 TI - Sudden gains in cognitive behavioral treatment for depression: when do they occur and do they matter? AB - Despite the use of efficacious treatments for depression, individuals differ in both the degree to which they recover and the rate at which recovery occurs. Tang and colleagues found that depressed patients who had sudden improvements in their symptomatology not only maintained these gains, but also enjoyed more improvement and higher rates of recovery than those without sudden gains (J. Consulting Clin. Psychol. 67(6) (1999) 894; J. Consulting Clin. Psychol. 70(2) (2002) 444). Our study examined the role of sudden gains in a cognitive-behavioral group treatment for depression. Results indicated that 41.9% of patients experienced sudden gains. Furthermore, sudden gains occurring in the first third of treatment appear to have special importance. Participants enjoying early sudden gains had significantly larger changes in depressive symptom scores over the course of treatment than those without sudden gains and were marginally more likely to be treatment responders compared to those without early sudden gains. In contrast to Tang and DeRubeis (J. Consulting Clin. Psychol. 67(6) (1999) 894), however, sudden gains were not associated with cognitive changes. PMID- 15890165 TI - Generalizability of carry-over effects in the emotional Stroop task. AB - The emotional Stroop task has been the most widely used task to examine attentional bias to emotionally salient stimuli. In one format of this task, words are presented to participants in a mixed randomized or quasi-randomized sequence. Using a mixed smoking Stroop task, we have previously demonstrated that smokers are slower to respond to words which follow smoking-related words than words which follow neutral words. Here we show that this carry-over effect is present in heroin addicts-but not control subjects-in a heroin Stroop task, and in normal subjects in a stress Stroop task. Thus, the effect generalizes to other populations. In addition, an examination of the studies that have collected data from both mixed and blocked formats provides converging evidence for the presence of carry-over effects. We discuss the implications of the carry-over effect for research using the emotional Stroop task. PMID- 15890166 TI - A psychometric evaluation of the social skills rating system in children with attention deficit hyperactivity disorder. AB - The social skills rating system (SSRS) was developed to assess social skills of children, as observed by multiple raters (teacher, parent, child). Studies of the SSRS have been conducted with handicapped, mentally retarded and learning disabled children. No studies have reported the psychometric properties of the SSRS in a clinical ADHD sample. This is important, because deficient social functioning is associated with ADHD. The present study assesses the psychometric properties of the teacher, parent and child versions of the SSRS in children with ADHD (n = 123), and normal controls (n = 239). Also, the social skills of children with ADHD, as rated on the SSRS were examined. Results support the factor structure and internal consistency of the original SSRS-teacher version. Moreover, support was found for 3 out of 4 scales of the SSRS-parent version. The factor structure of the SSRS-child version could not be replicated. An explanatory factor analysis on the SSRS-child version yielded two factors. Evidence was found for discriminative ability of the SSRS between normal controls and children with ADHD. Finally, informant agreement between raters was found to be poor. PMID- 15890167 TI - Do blushing phobics overestimate the undesirable communicative effects of their blushing? AB - Previous research indicated that blushing has socially threatening revealing effects in ambiguous situations. To explain blushing phobics' fearful preoccupation with blushing, we tested the hypothesis that blushing fearful individuals overestimate its revealing effects. High (n = 20) and low (n = 20) blushing fearful individuals read vignettes describing prototypical mishaps and ambiguous social events. Participants were prompted in the perspective of the actor, and were asked to indicate their expectations of the observers' judgments (meta-perceptions). Blushing fearful individuals overestimated the probability and the costs of undesirable outcomes. However, this judgmental bias was not inflated by displaying a blush. Thus, the results provide no evidence to support the idea that fear of blushing is fuelled by a biased conception of its communicative effects. PMID- 15890168 TI - Social interpretation bias and generalized social phobia: the influence of developmental experiences. AB - Patients with generalized social phobia (N = 42) and non-phobic community controls (N = 42) engaged in a social interaction with an experimental assistant whose behavior was used to create either a positive or an ambiguous social environment. Participants then rated their own performance and their partner's behavior. As a group, social phobic patients displayed negatively biased self judgments, but failed to display biased social interpretations. Among the social phobia group, a social developmental history marked by parental hostility was associated with negative interpretations of partner behavior and a history of parental overprotection was associated with less sensitivity to partner behavior. The results supported cognitive models of social phobia, which implicate negative learning experiences in the development of information processing biases. PMID- 15890169 TI - Enduring consequences of experimentally induced biases in interpretation. AB - Previous research has demonstrated that it is possible to induce biases in the interpretation of ambiguous text passages by training. Participants consistently trained to interpret emotionally ambiguous passages in either a negative or positive direction show training-congruent effects when presented with new ambiguous material. These training effects are demonstrated by participants' subsequent recognition ratings for disambiguating sentences, which represent both possible meanings of the novel ambiguous test passages. In this series of experiments, we investigated the durability of these training effects over time spans of up to 1 day and found them to be robust. The findings encourage us to believe that induced biases may serve as a useful analogue to those observed clinically. PMID- 15890170 TI - A cross-sectional evaluation of the factorial invariance of anxiety sensitivity in adolescents and young adults. AB - This study examined the cross-sectional factorial invariance of anxiety sensitivity in an ethnically diverse sample of adolescents (n = 173; mean age 15.5 years) and young adults (n = 291; mean age 20.1 years). Research in adult and youth samples suggests that anxiety sensitivity is best understood as a hierarchical construct with several lower-order factors. Factor models based on previous research using both adult and youth samples were compared and a hierarchical model with three lower-order factors provided the best fit to the data. Results supported the hypothesis that the factor structure of the Anxiety Sensitivity Index was invariant across age and gender. The factor scores also demonstrated differential correlations with symptoms of anxiety and depression. Results are discussed with regard to construct validation and understanding the structure of anxiety sensitivity in youth. PMID- 15890171 TI - Experimental modification of beliefs in obsessive-compulsive disorder: a test of the metacognitive model. AB - Metacognitive beliefs about the threatening meaning and significance of intrusions are fundamental to the development and maintenance of obsessive compulsive disorder (OCD) in the metacognitive model (Cognitive therapy of anxiety disorders: a practice manual and conceptual guide, Chichester, UK, Wiley, 1997; Emotional disorders and metacognition: innovative cognitive therapy, Chichester, UK, Wiley, 2000). This predicts that reductions in metacognitive beliefs about intrusions will result in decreased anxiety and compulsions. It was hypothesized that brief (5min) exposure and response prevention configured as a behavioural experiment to challenge metacognitive beliefs would lead to reductions in anxiety, thought fusion beliefs and the urge to neutralize. Furthermore, these reductions would be greater than the effects observed in the same exposure and response prevention task, accompanied by a habituation rationale. The results were consistent with the hypotheses. The clinical implications for the treatment of OCD are discussed. PMID- 15890172 TI - Global processing speed as a mediator of developmental changes in children's auditory memory span. AB - This study examined the role of global processing speed in mediating age increases in auditory memory span in 5- to 13-year-olds. Children were tested on measures of memory span, processing speed, single-word speech rate, phonological sensitivity, and vocabulary. Structural equation modeling supported a model in which age-associated increases in processing speed predicted the availability of long-term memory phonological representations for redintegration processes. The availability of long-term phonological representations, in turn, explained variance in memory span. Maximum speech rate did not predict independent variance in memory span. PMID- 15890173 TI - The cognitive foundations of reading and arithmetic skills in 7- to 10-year-olds. AB - A range of possible predictors of arithmetic and reading were assessed in a large sample (N=162) of children between ages 7 years 5 months and 10 years 4 months. A confirmatory factor analysis of the predictors revealed a good fit to a model consisting of four latent variables (verbal ability, nonverbal ability, search speed, and phonological memory) and two manifest variables (digit comparison and phoneme deletion). A path analysis showed that digit comparison and verbal ability were unique predictors of variations in arithmetic skills, whereas phoneme deletion and verbal ability were unique predictors of variations in reading skills. These results confirm earlier findings that phoneme deletion ability appears to be a critical foundation for learning to read (decode). In addition, variations in the speed of accessing numerical quantity information appear to be a critical foundation for the development of arithmetic skills. PMID- 15890174 TI - Representation and working memory in early arithmetic. AB - Working memory has been implicated in the early acquisition of arithmetic skill, but the relations among different components of working memory, performance on different types of arithmetic problems, and development have not been explored. Preschool and Grade 1 children completed measures of phonological, visual spatial, and central executive working memory, as well as nonverbal and verbal arithmetic problems, some of which included irrelevant information. For preschool children, accuracy was higher on nonverbal problems than on verbal problems, and the best and only unique predictor of performance on the standard nonverbal problems was visual-spatial working memory. This finding is consistent with the view that most preschoolers use a mental model for arithmetic that requires visual-spatial working memory. For Grade 1 children, performance was equivalent on nonverbal and verbal problems, and phonological working memory was the best predictor of performance on standard verbal problems. For both age groups, problems with added irrelevant information were substantially more difficult than standard problems, and in some cases measures of the central executive predicted performance. Assessing performance on different components of working memory in conjunction with different types of arithmetic problems provided new insights into the developing relations between working memory and how children do arithmetic. PMID- 15890175 TI - Age changes in the missing-letter effect revisited. AB - When participants search for a target letter while reading, they make more omissions if the target letter is embedded in frequent function words than in less frequent content words. Reflecting developmental changes in component language and literacy skills, the size of this effect increases with age. With adults, the missing-letter effect is due to both word function and word frequency. With children, it is unclear whether the growing size of the missing letter effect across development is due to a larger effect of word function, word frequency, or both because previous studies with children seeking to isolate the influence of word frequency and word function suffer from important methodological limitations. With these methodological limitations eliminated (Experiments 1 and 2), performance in a letter detection task was assessed for children in Grades 1, 2, 3, 4, and 7 as well as for undergraduate students. The results revealed that the influence of word function increases with age, whereas the effect of frequency is fairly stable across ages. Furthermore, normative predictability data collected in Experiment 3 revealed that third graders and undergraduate students were equally good at predicting function slots in a sentence. PMID- 15890176 TI - Recent progress in juvenile hormone research. Proceedings of the Eighth International Conference on Juvenile Hormones. June 20-24, 2004. King's Beach, California, USA. PMID- 15890177 TI - Immunocytochemical localization of an allatotropin in developmental stages of Heliothis virescens and Apis mellifera. AB - Juvenile hormone biosynthesis by the corpora allata is regulated by stimulatory neuropeptides called allatotropins and inhibitory neuropeptides called allatostatins. This study localized Manduca sexta allatotropin-like material in developmental stages of the noctuid moth Heliothis virescens and the honeybee Apis mellifera. Immunocytochemical methods using both fluorescence-tagged antibodies and enzyme-coupled antibodies were used to stain the central nervous tissue of both species. H. virescens contains M. sexta allatotropin (Manse-AT) like material consistently throughout larval development. The distribution patterns of Manse-AT immunoreactive cell bodies in the CNS persisted from one larval instar to the next. It will be discussed how larval Manse-AT distribution patterns differed from those in adults. The total number of AT-containing cells in brain and subesophageal ganglion gradually increased during larval development, whereas in the thoracic and abdominal ganglia, the number of AT containing neurons remained constant. In the honeybee A. mellifera, Manse-AT immunoreactive cells were only found in a few brains from late last instar larvae (prepupae). Manse-AT-like material was present in a group of 6-8 cells in the pars intercerebralis. However, we did not find any Manse-AT-like material in brains of early last instar larvae, whose corpora allata (CA) are more sensitive to in vitro stimulation by Manse-AT than prepupal CA. PMID- 15890178 TI - Phe-Gly-Leu-amide allatostatin in the termite Reticulitermes flavipes: content in brain and corpus allatum and effect on juvenile hormone synthesis. AB - In the subterranean termite Reticulitermes flavipes, allatostatins (ASTs) with the C-terminus Phe-Gly Leu-amide were localized by immunocytochemistry with antibody against a cockroach AST, Dippu AST-7. AST-immunoreactivity occurred in the corpus cardiacum and corpus allatum and in the lateral and medial neurosecretory cells of the brain that innervate these organs as well as in many other nerve cells of the brain. This was observed in workers, nymphs, soldiers and secondary reproductives. A radioimmunoassay, using anti-Dippu AST-11, demonstrated about 40 fmole equivalents of AST in brains of soldiers and secondary reproductives. The product of the corpora allata in this species was determined to be juvenile hormone III. Its synthesis by corpora allata of secondary reproductives, determined by in vitro radiochemical assay, was inhibited in a dose-dependent fashion by two cockroach allatostatins, Dippu AST-7 and Dippu AST-11. Thus, as in cockroaches and crickets, allatostatin-containing nerves innervate the corpora allata of this termite species and their production of juvenile hormone is inhibited by these neuropeptides. PMID- 15890179 TI - Regulation of methyl farnesoate production by mandibular organs in the crayfish, Procambarus clarkii: a possible role for allatostatins. AB - Decapod crustaceans do not appear to produce juvenile hormone, but rather its immediate precursor, methyl farnesoate (MF). Both MF and its immediate precursor, farnesoic acid (FA) are produced by the mandibular organs (MO) in crustaceans. The MO are homologous to the insect corpora allata (CA), the site of juvenile hormone biosynthesis. However, the FGLamide allatostatin (ASTs) peptides, of which there are about 60 distinct forms reported from crustaceans, have previously been found to have no effect on MO activity in crustaceans. We have identified by immunocytochemistry the presence of FGLamide-like AST immunoreactivity in neurosecretory cells throughout the CNS as well as in neurohaemal structures such as the sinus gland and pericardial organs. The ASTs are likely delivered to the MO hormonally and/or by local neurohaemal release. Using MO from adult males, we have found wide variability between animals in the in vitro rates of MF and FA biosynthesis. Treatment with Dippu-ASTs has a statistically significant stimulatory effect on MF synthesis, but only in MO that are initially producing MF at lower rates. No effect on FA production was observed, suggesting that the FGLamide ASTs exert their effect on the o-methyl transferase, the enzyme responsible for the conversion of FA to MF. PMID- 15890180 TI - Methyl farnesoate controls adult male morphogenesis in the crayfish, Procambarus clarkii. AB - Adult male crayfish Procambarus clarkii exist in two morphotypes. They continue to molt as adults, switching between Form Is and Form IIs. Form Is are primary reproductive types, with large chelae and spines on the ischiopodites of the third and fourth pair of walking legs. Form IIs are non-reproductive types with smaller chelae and no spines on the ischiopodites. We investigated the hormonal control of these transitions in two ways, by eyestalk ablation and by methyl farnesoate (MF) treatments. Eyestalk ablation accelerates molting and increases MF levels in the blood. MF is a hormone that regulates both reproduction and morphogenesis. MF concentrations were determined in two ways. The hemolymph samples were extracted first, then purified, using normal phase HPLC. The fractions containing MF were collected and analyzed for MF concentration, utilizing both internal and external standards by GC/MS. The other hemolymph samples were analyzed from individual animals by HPLC. The concentrations of ecdysteroids were determined by radioimmunoassay. In the control animals, 4 out of 4 untreated Form I males molted into Form II, while 6 out of 7 Form IIs molted into Form Is. Eight of 8 ablated Form Is molted into Form IIs as expected, while 5 of 5 ablated Form IIs molted into Form IIs, instead of Form Is. MF treatment of intact animals resulted in 6 of 7 Form Is becoming Form IIs and 5 of 6 Form IIs becoming Form IIs. These results were highly significant in comparison of Form I and IIs in each treatment (eyestalk intact, eyestalk ablated and eyestalk intact with MF) by a chi square analysis, P = 0.006, P < 0.0005, and P = 0.013, respectively. MF premolt blood levels suggested that Form IIs were produced in the presence of 1.3 ng/ml MF, while Form Is result from MF levels less than 0.5 ng/ml. Since both eyestalk ablation and MF treatment resulted in the failure of Form IIs becoming Form Is, it was concluded that the control of morphogenesis of primary reproductives (Form Is) depends on a low level of MF prior to the molt, while Form IIs are formed in the presence of increased levels of MF. PMID- 15890181 TI - Regulation of juvenile hormone titers by soldiers in the Formosan subterranean termite, Coptotermes formosanus. AB - In field collections of the Formosan subterranean termite Coptotermes formosanus, soldiers averaged less than 10%. The proportion of soldiers increased to about 25% or higher when termites were kept in the laboratory, as did the juvenile hormone III (JH III) titers for both workers and soldiers. In laboratory experiments with a proportion of soldiers 25% or higher, very few new pre soldiers were formed and the JH titer in existing soldiers remained constant. On the contrary, workers from groups containing less than 25% soldiers formed more pre-soldiers. In such cases, both workers and soldiers showed higher JH titers. Newly formed soldiers also had higher JH titers than older soldiers. It is speculated that populations containing higher soldier proportions inhibit further soldier differentiation, eventually holding the JH titers under the threshold or suppressing the activity of corpora allata resulting in lower JH titers in workers. PMID- 15890182 TI - Developmental and hormonal regulation of juvenile hormone esterase gene in Drosophila melanogaster. AB - Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was used to study developmental expression and hormonal regulation of the juvenile hormone esterase gene (DmJhe) in the fruit fly, Drosophila melanogaster. The levels of DmJhe mRNA were low during the embryonic stage. A peak of Dmjhe mRNA was detected in the first, second and third instar larvae. The Dmjhe mRNA levels also increased soon after pupal ecdysis. The Dmjhe mRNA was detected in both male and female adult flies. The peaks of Dmjhe mRNA observed in the larvae coincided with the peaks of juvenile hormone (JH). In contrast, the mRNA for ecdysone-induced transcription factor, Drosophila hormone receptor 3 (DHR3) showed peaks of expression that coincided with the ecdysteroid peaks in embryo, larva and pupa. JH III induced Dmjhe mRNA but not DHR3 mRNA in explanted tissues cultured in Grace's medium. 20-hydroxyecdysone induced DHR3 mRNA and suppressed JH III induction of DmJhe mRNA. These studies show that the expression of jhe in D. melanogaster is regulated by both JH and 20E. PMID- 15890183 TI - The effect of the juvenile hormone analog, fenoxycarb on the PBAN-receptor and pheromone production in adults of the moth Helicoverpa armigera: an "aging" hormone in adult females? AB - In a previous study we showed that juvenile hormone (JH) or its analog, fenoxycarb (FX), is involved in the up-regulation of pheromone biosynthesis activating neuropeptide (PBAN) competence. JH causes induction of binding to a putative PBAN-receptor (PBAN-R) and the subsequent pheromone production by pheromone glands of pharate females. The present study demonstrates that pheromone production by the adult female is age-dependent. The pheromonotropic response increased to reach a maximum at 4 days, after which a decreased response was observed. Binding of the PBAN-R was also age-dependent. Treatment with FX inhibited both binding of PBAN to the PBAN-R and the pheromonotropic response as reflected by the production of the main pheromone component, Z-11-hexadecenal. Thus, in contrast to its up-regulatory role in pharate females, FX treatment of adult females causes down-regulation of both pheromone production and specific binding to the PBAN-R. In addition, behavioural observations showed that calling behaviour, mating success and subsequent egg-fertility are affected by treating females with FX. PMID- 15890184 TI - Effect of precocene II on fatty acid metabolism in the pea aphid, Acyrthosiphon pisum, under cold stress. AB - Pea aphids, Acyrthosiphon pisum (Harris), reared at 10 degrees C contain higher levels of fatty acids than those reared at 25 degrees C. This is primarily the result of an accumulation of triacylglycerols containing myristic acid. When aphids reared at 25 degrees C were transferred to 10 degrees C there was a gradual increase in triacylglycerol content that reached a maximum at 16 days post-transfer. Treatment of aphids with precocene II prior to transfer to 10 degrees C blocked the accumulation of fatty acids including myristic acid. A single application of 2 microg precocene II/aphid or two applications of 0.5 microg precocene II/ aphid administered on consecutive days resulted in aphids moved to 10 degrees C maintaining the same fatty acid profile as aphids maintained at 25 degrees C. Aphids that were treated with precocene II and maintained at 25 degrees C did not show changes in fatty acid profiles. Rearing aphids at 10 degrees C resulted in lower rates of reproduction and lower total numbers of progeny with longer longevity. Treatment with precocene II significantly decreased the total number of progeny produced at both temperatures. Precocene II did not reduce life span of aphids reared at 25 degrees C, however, the life span of treated aphids reared at 10 degrees C was decreased. The mechanism by which precocene II prevents the accumulation of myristic acid in aphids reared at 10 degrees C remains to be determined. PMID- 15890185 TI - Juvenile hormone, 20-hydroxyecdysone and dopamine interaction in Drosophila virilis reproduction under normal and nutritional stress conditions. AB - To elucidate the role of the juvenile hormone (JH) in the control of Drosophila reproduction under stress, JH degradation, dopamine (DA) content and reproduction were studied upon 20E treatment in Drosophila virilis females of wild type (wt) and a mutant, with increased 20E level and decreased fertility, under normal and nutritional stress conditions. 20E treatment of wt flies for 7 days results in an increase of DA content in young females, but a decrease in mature females, a decrease of JH degradation in both young and mature females, an 1-day delay in onset of oviposition and a decrease of fecundity to the level typical of mutant flies. One day of 20E treatment in 7-day-old fed and starved flies results in a small decrease of JH degradation in the fed females and a great decrease in the starved ones. Fecundity decreases in the fed flies to the levels of the starved untreated flies in both wt and mutant strains. An oviposition arrest is observed in the treated and the untreated starved, but not in the treated fed, females of both strains. The data obtained suggest ecdysone control of JH metabolism mediated via DA. PMID- 15890186 TI - JH III production, titers and degradation in relation to reproduction in male and female Anthonomus grandis. AB - Juvenile hormone (JH) is necessary for the production of vitellogenin (Vg) in the boll weevil, Anthonomus grandis. Occurrence of Vg in this species is typically restricted to reproductively competent females, and is not detected in untreated males. However, the JH analog, methoprene stimulates Vg production in intact males and in the isolated abdomens of both male and female boll weevils (where in each case no Vg is detected without treatment), suggesting that males are competent to produce Vg but are normally not stimulated to do so. Preliminary work indicating that male boll weevil corpora allata (CA) produced little or no JH in vitro suggested that failure of males to produce Vg might be due to very low JH levels compared to females. This study re-examines the question of JH in male boll weevils by determining in vitro production of JH III by male CA during the first 10 days after adult emergence, determining hemolymph JH esterase activity during this same time period and hemolymph JH III titers in adults of both sexes. We also re-examine the ability of isolated male abdomens to produce Vg in response to hormonal stimulation, analyzing the effect of a wide range of methoprene and JH III dosages. Results indicate that male A. grandis have circulating JH titers and JH production similar to females. JH esterase activity is slightly but significantly higher in males than females. Vg production by isolated abdomens of both sexes after stimulation with methoprene or JH III was confirmed. Dose response studies indicated that high levels of methoprene were less effective than intermediate doses in stimulating Vg synthesis in both sexes. We conclude that the sexually dimorphic effect of JH on Vg synthesis is not due to differences in JH production or differences in JH titer between the sexes. PMID- 15890187 TI - Starvation affects vitellogenin production but not vitellogenin mRNA levels in the lubber grasshopper, Romalea microptera. AB - The interaction of juvenile hormone (JH) and nutrition was studied during the oviposition cycle of the Eastern Lubber grasshopper (Romalea microptera). Starvation of females early or in the middle of the cycle inhibited oocyte growth. Starvation for 4 days also reduced hemolymph levels of JH III and vitellogenesis (Vg) to 25% and 15%, respectively, of the levels in fed animals. Likewise, Vg production by fat body fragments incubated in vitro was reduced to 2% of the levels in fed animals and total protein synthesis was reduced to 25%, suggesting that starvation had a stronger effect on Vg synthesis than on protein synthesis. These effects were reversed when starved animals were fed again. However, fat body levels of Vg-mRNA were similar in fed and starved animals, indicating that starvation did not affect transcript levels. We tested whether the decline in JH levels mediated the other starvation effects by infusing animals with JH III or vehicle for 2 days at the onset of starvation. Infusion of JH elevated JH and Vg-mRNA levels 670% and 103%, respectively, above the levels in vehicle-infused animals. However, Vg production and hemolymph levels of Vg were similar to the levels in vehicle-infused animals. These data suggest that JH alone is insufficient to stimulate Vg production. PMID- 15890188 TI - Pharmacological analysis of ovarial patency in Heliothis virescens. AB - The insect oocyte sequesters nutritive proteins during patency, which is facilitated as a result of intercellular spaces occurring between follicular epithelial cells under the influence of juvenile hormone (JH). Patency was analyzed in the moth, Heliothis virescens, using a pharmacological approach, in which we used different JH homologues and chemicals that specifically target elements of two second-messenger pathways in vertebrates, the cAMP-dependent and inositol triphosphate/diacylglycerol signaling pathways. JH I and JH III evoked dose-dependent patency in H. virescens oocyte follicles, which was suppressed by the Na/K-ATPase inhibitor, ouabain. Patency was observed in follicular epithelial cells treated with either protein kinase C activator, PDBu, or protein kinase A activator, 8-Br-cAMP, by itself. The protein kinase C inhibitor, H-7, preferentially suppressed patency evoked by JH III, whereas the protein kinase A inhibitor, H89, preferentially suppressed that evoked by JH I. Additionally, patency was triggered by the adenylate cyclase activator, NKH 477, or peptide Gs protein activator, cholera toxin, alone. Patency evoked by JH I was suppressed by the adenylate cyclase inhibitor, SQ 22,536, and GPAnt-2, a peptide antagonistic to Gs proteins that stimulates adenylate cyclase. Neither of these latter inhibitors, however, affected JH III-evoked patency. These results suggest that, in the process of patency in H. virescens ovarial follicles, JH I predominantly signals via the cAMP-dependent second messenger system, whereas JH III acts via the inositol triphosphate/diacylglycerol signaling pathway. Moreover, stimulation of patency by cholera toxin alone and inhibition of JH I-evoked patency by GPAnt 2, strongly suggest that JH I acts on the follicular epithelial cells via activation of G-protein, and-possibly-via G(s)-protein coupled receptor. PMID- 15890189 TI - Insulin signaling is necessary for vitellogenesis in Drosophila melanogaster independent of the roles of juvenile hormone and ecdysteroids: female sterility of the chico1 insulin signaling mutation is autonomous to the ovary. AB - It has been suggested that insulin signaling mutations of Drosophila melanogaster are sterile and long-lived because of juvenile hormone (JH) and ecdysteroid deficiency. However, female sterility of an insulin/IGF-like signaling mutant (chico(1)) of D. melanogaster is not mediated by downstream systemic signaling in terms of major alterations in JH or ecdysteroid levels. chico(1) is a null mutation in the insulin substrate protein (CHICO) gene of D. melanogaster. Homozygous chico(1) females are sterile and their oocytes do not mature beyond the last previtellogenic stage. Homozygous chico(1) females exhibit approximately wild-type rates of JH biosynthesis, ovarian release of ecdysteroids and haemolymph ecdysteroid levels, suggesting that these two major hormone systems play no role in producing the sterility. Previtellogenic wild-type ovaries transplanted into homozygous chico(1) females underwent vitellogenesis, showing that systemic factors present in mutant females are sufficient to support normal vitellogenesis. chico(1) ovaries transplanted into wild-type females did not undergo vitellogenesis indicating that CHICO is necessary in the ovary for vitellogenic maturation. The ovary transplant experiments corroborate the endocrine results and demonstrate that insulin/insulin-like signaling (IIS) is necessary for vitellogenesis even when sufficient levels of JH, ecdysteroids or other factors are present. PMID- 15890190 TI - The cockroach Leucophaea maderae needs more than juvenile hormone, vitellogenin and reserves to make a yolky egg. AB - For the cockroach Leucophaea maderae the developmental profile of lipophorin (Lp) concentrations in the hemolymph was determined through the entire vitellogenic period. At mid-vitellogenesis the concentrations of Lp had risen to 6 times the level at emergence and then declined to 2/3 of such high values at ovulation. The racemic 10R,10S-JH-III bound to Lp with an affinity of K(d) = 5.76 nM and the natural enantiomer 10R-JH-III with a K(d) = 1.60 nM. Injections of anti-Lp into mated females caused a significantly reduced rate of oocyte growth and a substantial degree of oosorption. Injections of gamma-globulin did not significantly reduce oocyte growth and caused only a small number of oocytes to resorb. Starvation after mating had similar effects as treatment with anti-Lp. Because of the high affinity of JH to Lp and since Lp occurs in micromolar concentrations during vitellogenesis one can assume that practically all JH is bound and not available for hydrolysis by the JH esterases. Lp appears to function as an inhibitor of JH metabolism by the JHEs through substrate depletion. One may conclude that a normal rate of egg growth is only achieved when titers of Lp exceed those of JH and remove major portions of this substrate from degradation by the JHEs. PMID- 15890191 TI - Presence and titer of methyl palmitate in the Medfly (Ceratitis capitata) during reproductive maturation. AB - The relative amounts of methyl palmitate (MP) during the first 10 days post eclosion were determined in whole-body extracts of adult female Ceratitis capitata by SIM monitoring of the 74 m/z fragment. MP peaks in receptive 3-day old virgin females coincide with previously reported production of Juvenile Hormone (JH) by the corpus allatum (CA). Mating in the Medfly induces female non receptivity. Indirect evidence suggests that the mevalonate pathway to sesquiterpene biosynthesis is underdeveloped in newly eclosed females. We propose that the pathway leading to synthesis of JH is markedly diverted in non-receptive virgin females to fatty acid synthesis, and partly so-in non-receptive mated females, leading to production of palmitic acid, presumably methylated thereafter. MP is depressed and remains marginal thereafter for the 7 days examined in the virgin female but goes through an apparent second cycle in the mated female. This contrasts with the consistent increase of allatal biosynthesis of MP of virgin and mated females previously reported and suggests additional control mechanisms in vivo. During the period of reduced receptivity following the first mating a second apparent peak of MP is observed. MP is a metabolic default metabolite of reproductively immature females whose putative role in reproductive physiology remains to be defined. PMID- 15890192 TI - L1-mediated retrotransposition of murine B1 and B2 SINEs recapitulated in cultured cells. AB - SINEs are short interspersed nucleotide elements with transpositional activity, present at a high copy number (up to a million) in mammalian genomes. They are 80 400 bp long, non-coding sequences which derive either from the 7SL RNA (e.g. human Alus, murine B1s) or tRNA (e.g. murine B2s) polymerase III-driven genes. We have previously demonstrated that Alus very efficiently divert the enzymatic machinery of the autonomous L1 LINE (long interspersed nucleotide element) retrotransposons to transpose at a high rate. Here we show, using an ex vivo assay for transposition, that both B1 and B2 SINEs can be mobilized by murine LINEs, with the hallmarks of a bona fide retrotransposition process, including target site duplications of varying lengths and integrations into A-rich sequences. Despite different phylogenetic origins, transposition of the tRNA derived B2 sequences is as efficient as that of the human Alus, whereas that of B1s is 20-100-fold lower despite a similar high copy number of these elements in the mouse genome. We provide evidence, via an appropriate nucleotide substitution within the B1 sequence in a domain essential for its intracellular targeting, that the current B1 SINEs are not optimal for transposition, a feature most probably selected for the host sake in the course of evolution. PMID- 15890193 TI - The human peroxisome proliferator-activated receptor delta gene is a primary target of 1alpha,25-dihydroxyvitamin D3 and its nuclear receptor. AB - Peroxisome proliferator-activated receptor (PPAR) delta is the most widely expressed member of the PPAR family of nuclear receptor fatty acid sensors. Real time PCR analysis of breast and prostate cancer cell lines demonstrated that PPARdelta expression was increased 1.5 to 3.2-fold after three hours stimulation with the natural vitamin D receptor (VDR) agonist, 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3). In silico analysis of the 20 kb of the human PPARdelta promoter revealed a DR3-type 1alpha,25(OH)2D3 response element approximately 350 bp upstream of the transcription start site, which was able to bind VDR-retinoid X receptor (RXR) heterodimers and mediate a 1alpha,25(OH)2D3-dependent upregulation of reporter gene activity. Chromatin immuno-precipitation assays demonstrated that a number of proteins representative for 1alpha,25(OH)2D3 mediated gene activation, such as VDR, RXR and RNA polymerase II, displayed a 1alpha,25(OH)2D3-dependent association with a region of the proximal PPARdelta promoter that contained the putative DR3-type VDRE. This was also true for other proteins that are involved in or are the subject of chromatin modification, such as the histone acetyltransferase CBP and histone 4, which displayed ligand dependent association and acetylation, respectively. Finally, real-time PCR analysis demonstrated that 1alpha,25(OH)2D3 and the synthetic PPARdelta ligand L783483 show a cell and time-dependent interference in each other's effects on VDR mRNA expression, so that their combined application shows complex effects on the induction of VDR target genes, such as CYP24. Taken together, we conclude that PPARdelta is a primary 1alpha,25(OH)2D3-responding gene and that VDR and PPARdelta signaling pathways are interconnected at the level of cross-regulation of their respective transcription factor mRNA levels. PMID- 15890194 TI - Spontaneous ribosome bypassing in growing cells. AB - Translating ribosomes can pass through a stretch of messenger RNA without translating and resume protein chain elongation after the bypassed region. We previously investigated the stimulation of bypassing when the codon in the ribosome [corrected] A-site called for an aminoacyl-tRNA species in short supply. Here, we investigate bypassing in unstarved, growing cells. A collection of lacZ bypass reporters was constructed with nearly all the sense codons as the "takeoff site", each with its matched landing site 16 nucleotides downstream in the beta galactosidase reading frame. Beta-galactosidase [corrected] synthesis in unstarved cells carrying these reporters was found to vary over a large range. The takeoff sites UUU and AGG yielded unusually high enzyme activities, sufficient for protein sequence analysis; in these cases, sequencing (by Edman degradation or by mass spectrometry) confirmed that the synthesis of lacZ protein occurred through the 16 nt bypass from takeoff to landing site. Thus, bypassing occurs spontaneously under normal conditions, and is not limited to the pathology of amino acid starvation. Indirect evidence suggests that most of the lower enzyme activities of the rest of the collection also reflects bypassing. Another collection of reporters was made with [corrected] various triplets in the A-site [corrected] the codon immediately following a UUC [corrected] takeoff triplet. Spontaneous bypassing in representatives of this collection varied roughly inversely with the abundance of the tRNA encoded at the A-site. For two A-site codons tested, introduction of additional copies of the relevant tRNA gene on a second plasmid reduced spontaneous bypassing. We conclude that any pause with the A-site empty stimulates bypassing. From the P-site and A-site effects on bypassing, we estimated the average frequency of ribosome takeoff; from this, we calculate that the probability that a ribosome will succeed in translating the entire lacZ coding sequence is about 0.73, in agreement with earlier, independent estimates. PMID- 15890195 TI - Structural transitions induced by the interaction between tRNA(Gly) and the Bacillus subtilis glyQS T box leader RNA. AB - The T box system regulates expression of amino acid-related genes in Gram positive bacteria through premature termination of transcription. Synthesis of the full-length mRNA requires stabilization of an antiterminator element in the 5' untranslated leader RNA by the cognate uncharged tRNA. tRNA(Gly)-dependent antitermination of the Bacillus subtilis glyQS gene (encoding glycyl-tRNA synthetase) can be reproduced in a purified in vitro transcription system, indicating that the nascent transcript is sufficient for interaction with the tRNA. Genetic analyses previously demonstrated base pairing of a single codon in the leader RNA with the tRNA anticodon, and between the antiterminator and the tRNA acceptor end. In this study, we established conditions for specific binding of tRNA(Gly) to glyQS leader RNA generated by phage T7 RNA polymerase. Structural mapping studies revealed tRNA(Gly)-induced protection in the glyQS leader RNA at the two known sites of interaction with the tRNA, as well as at other regions between these sites. The proposed tRNA-dependent structural switch between the competing terminator and antiterminator forms of the leader RNA was demonstrated directly. Changes in tRNA(Gly) upon binding to glyQS leader RNA were detected in the anticodon loop, consistent with pairing with the specifier sequence, and in the highly conserved G19 in the D-loop, similar to effects induced by codon anticodon interaction in the ribosome. This study provides biochemical evidence for direct interaction of tRNA(Gly) with full-length in vitro transcribed glyQS leader RNA, and an initial view of structural modulations of both RNA partners within the complex. PMID- 15890196 TI - Non-bridging phosphate oxygen atoms within the tRNA anticodon stem-loop are essential for ribosomal A site binding and translocation. AB - The conformation of the anticodon stem-loop of tRNAs required for correct decoding by the ribosome depends on intramolecular and intermolecular interactions that are independent of the tRNA nucleotide sequence. Non-bridging phosphate oxygen atoms have been shown to be critical for the structure and function of several RNAs. However, little is known about the role they play in ribosomal A site binding and translocation of tRNA to the P site. Here, we show that non-bridging phosphate oxygen atoms within the tRNA anticodon stem-loop at positions 33, 35, and 37 are important for A site binding. Those at positions 34 and 36 are not necessary for binding, but are essential for translocation. Our results correlate with structural data, indicating that position 34 interacts with the highly conserved 16S rRNA base G966 and position 36 interacts with the universally conserved tRNA base U33 during translocation to the P site. PMID- 15890197 TI - Flexibility in repression and cooperativity by KorB of broad host range IncP-1 plasmid RK2. AB - KorB, encoded by plasmid RK2, belongs to the ParB family of active partitioning proteins. It binds to 12 operators on the RK2 genome and was previously known to repress promoters immediately adjacent to operators O(B)1, O(B)10 and O(B)12 (proximal) or up to 154 bp away (distal) from O(B)2, O(B)9 and O(B)11. To achieve strong repression, KorB requires a cooperative interaction with one of two other plasmid-encoded repressors, KorA or TrbA. Reporter gene assays were used in this study to test whether the additional KorB operators may influence transcription and to test how KorB acts at a distance. The distance between O(B)9 and trbBp could be increased to 1.6kb with little reduction in repression or cooperativity with TrbA. KorB was also able to repress the promoter and cooperate with TrbA when the O(B) site was placed downstream of trbBp. This suggested a potential regulatory role for O(B) sites located a long way from any known promoter on RK2. O(B)4, 1.9kb upstream of traGp, was shown to mediate TrbA-potentiated KorB repression of this promoter, but no effect on traJp upstream of O(B)4 was observed, which may be due to the roadblocking or topological influence of the nucleoprotein complex formed at the adjacent transfer origin, oriT. Repression and cooperativity were alleviated significantly when a lac operator was inserted between O(B)9 and trbBp in the context of a LacI+ host, a standard test for spreading of a DNA-binding protein. On the other hand, a standard test for DNA looping, movement of the operator to the opposite face of the DNA helix from the natural binding site, did not significantly affect KorB repression or cooperativity with TrbA and KorA over relatively short distances. While these results are more consistent with spreading as the mechanism by which KorB reaches its target, previous estimates of KorB molecules per cell are not consistent with there being enough to spread up to 1kb from each O(B). A plausible model is therefore that KorB can do both, spreading over relatively short distances and looping over longer distances. PMID- 15890198 TI - Salt dependent binding of T4 gene 32 protein to single and double-stranded DNA: single molecule force spectroscopy measurements. AB - Bacteriophage T4 gene 32 protein (gp32) is a well-studied representative of the large family of single-stranded DNA (ssDNA) binding proteins, which are essential for DNA replication, recombination and repair. Surprisingly, gp32 has not previously been observed to melt natural dsDNA. At the same time, *I, a truncated version of gp32 lacking its C-terminal domain (CTD), was shown to decrease the melting temperature of natural DNA by about 50 deg. C. This profound difference in the duplex destabilizing ability of gp32 and *I is especially puzzling given that the previously measured binding of both proteins to ssDNA was similar. Here, we resolve this apparent contradiction by studying the effect of gp32 and *I on the thermodynamics and kinetics of duplex DNA melting. We use a previously developed single molecule technique for measuring the non-cooperative association constants (K(ds)) to double-stranded DNA to determine K(ds) as a function of salt concentration for gp32 and *I. We then develop a new single molecule method for measuring K(ss), the association constant of these proteins to ssDNA. Comparing our measured binding constants to ssDNA for gp32 and *I we see that while they are very similar in high salt, they strongly diverge at [Na+] < 0.2 M. These results suggest that intact protein must undergo a conformational rearrangement involving the CTD that is in pre-equilibrium to its non-cooperative binding to both dsDNA and ssDNA. This lowers the effective concentration of protein available for binding, which in turn lowers the rate at which it can destabilize dsDNA. For the first time, we quantify the free energy of this CTD unfolding, and show it to be strongly salt dependent and associated with sodium counter-ion condensation on the CTD. PMID- 15890199 TI - Synapsis in phage Bxb1 integration: selection mechanism for the correct pair of recombination sites. AB - Recombination by site-specific recombinases is a highly concerted process that requires synapsis of the correct pair of DNA substrates. Phage-encoded serine integrases are unusual among the serine-recombinase family, which includes transposon resolvases and DNA invertases, in that they utilize two simple but different DNA substrates (attB and attP) and do not require accessory sites, additional proteins, or DNA supercoiling. Synapsis must therefore be directed solely by integrase-DNA interactions. We show here that the Bxb1 serine-integrase binds as a dimer to its two DNA substrates (attB, attP) and recombinant products (attL, attR) with similar affinities. However, synapsis occurs only between attP and attB, and not between any of the other nine possible site combinations. The Bxb1 integrase domain structure, the unusual DNA-binding properties of the integrase, and the characterization of a mutant protein with altered site discrimination, are consistent with synaptic selectivity being derived from DNA sequence-induced changes in the conformations of integrase-DNA complexes. PMID- 15890200 TI - The Ankrd2, Cdkn1c and calcyclin genes are under the control of MyoD during myogenic differentiation. AB - Skeletal muscle development requires the coordinated expression of numerous transcription factors to control the specification of the muscle fate in mesodermal cells and the differentiation of the committed myoblasts into functional contractile fibers. The bHLH transcription factor MyoD plays a key role in these processes, since its forced expression is sufficient to induce the myogenesis in a variety of non-muscle cells in culture. Consistent with this observation, the majority of skeletal muscle genes require MyoD to activate their own transcription. In order to identify novel MyoD-target genes we generated C2C12 MyoD-silenced clones, and used a muscle-specific cDNA microarray to study the induced modifications of the transcriptional profile. Gene expression was analyzed at three different stages in differentiating MyoD(-)C2C12 myoblasts. These microarray data sets identified many additional uncharacterized downstream MyoD transcripts that may play important functions in muscle cell differentiation. Among these genes, we concentrated our study on the cell cycle regulators Cdkn1c and calcyclin and on the muscle-specific putative myogenic regulator Ankrd2. Bioinformatic and functional studies on the promoters of these genes clarified their dependence on MyoD activity. Clues of other regulatory mechanisms that might interact with the principal bHLH transcription factor have been revealed by the unexpected up-regulation in MyoD(-) cells of these novel (and other) target transcripts, at the differentiation stage in which MyoD became normally down-regulated. PMID- 15890201 TI - Myomesin is a molecular spring with adaptable elasticity. AB - The M-band is a transverse structure in the center of the sarcomere, which is thought to stabilize the thick filament lattice. It was shown recently that the constitutive vertebrate M-band component myomesin can form antiparallel dimers, which might cross-link the neighboring thick filaments. Myomesin consists mainly of immunoglobulin-like (Ig) and fibronectin type III (Fn) domains, while several muscle types express the EH-myomesin splice isoform, generated by the inclusion of the unique EH-segment of about 100 amino acid residues (aa) in the center of the molecule. Here we use atomic force microscopy (AFM), transmission electron microscopy (TEM) and circular dichroism (CD) spectroscopy for the biophysical characterization of myomesin. The AFM identifies the "mechanical fingerprints" of the modules constituting the myomesin molecule. Stretching of homomeric polyproteins, constructed of Ig and Fn domains of human myomesin, produces a typical saw-tooth pattern in the force-extension curve. The domains readily refold after relaxation. In contrast, stretching of a heterogeneous polyprotein, containing several repeats of the My6-EH fragment reveals a long initial plateau corresponding to the sum of EH-segment contour lengths, followed by several My6 unfolding peaks. According to this, the EH-segment is characterized as an entropic chain with a persistence length of about 0.3nm. In TEM pictures, the EH domain appears as a gap in the molecule, indicating a random coil conformation similar to the PEVK region of titin. CD spectroscopy measurements support this result, demonstrating a mostly non-folded conformation for the EH-segment. We suggest that similarly to titin, myomesin is a molecular spring, whose elasticity is modulated by alternative splicing. The Ig and Fn domains might function as reversible "shock absorbers" by sequential unfolding in the case of extremely high or long sustained stretching forces. These complex visco-elastic properties of myomesin might be crucial for the stability of the sarcomere. PMID- 15890202 TI - Molecular-scale topographic cues induce the orientation and directional movement of fibroblasts on two-dimensional collagen surfaces. AB - Collagen fibres within the extracellular matrix lend tensile strength to tissues and form a functional scaffold for cells. Cells can move directionally along the axis of fibrous structures, in a process important in wound healing and cell migration. The precise nature of the structural cues within the collagen fibrils that can direct cell movement are not known. We have investigated the structural features of collagen that are required for directional motility of mouse dermal fibroblasts, by analysing cell movement on two-dimensional collagen surfaces. The surfaces were prepared with aligned fibrils of collagen type I, oriented in a predefined direction. These collagen-coated surfaces were generated with or without the characteristic 67 nm D-periodic banding. Quantitative analysis of cell morphodynamics showed a strong correlation of cell elongation and motional directionality with the orientation of D-periodic collagen microfibrils. Neither directed motility, nor cell body alignment, was observed on aligned collagen lacking D-periodicity, or on D-periodic collagen in the presence of peptide containing an RGD motif. The directional motility of fibroblast cells on aligned collagen type I fibrils cannot be attributed to contact guidance, but requires additional structural information. This allows us to postulate a physiological function for the 67 nm periodicity. PMID- 15890203 TI - VceR regulates the vceCAB drug efflux pump operon of Vibrio cholerae by alternating between mutually exclusive conformations that bind either drugs or promoter DNA. AB - VceR, a member of the TetR family of transcriptional regulators, is a repressor of the vceCAB operon, which encodes a multidrug efflux pump in Vibrio cholerae. VceR binds to a 28 bp inverted-repeat within the vceR-vceC intergenic region and is dissociated from this site with CCCP, a pump substrate. The rate of the CCCP induced conformational change in VceR was determined by stopped-flow fluorescence spectroscopy, revealing a highly co-operative process that occurs with a Hill coefficient of approximately 4. The apparent affinity for CCCP decreased in a linear manner with increasing concentrations of DNA, indicative of competition between the CCCP and DNA for binding to VceR. These data are consistent with an equilibrium between mutually exclusive conformations that are supported by the binding of DNA and CCCP to the N and C termini of VceR, respectively. Size exclusion chromatography and dynamic light-scattering studies indicate that VceR exists predominantly as a dimer; however, a pair of dimers binds to the DNA. In order to account for the fact that VceR is a dimer in the absence of DNA but binds CCCP with a Hill co-efficient of 4, implying that it has at least four binding-sites, we propose that the VceR monomer possesses a pair of binding sites that can be simultaneously occupied by CCCP. Using a gene-reporter system and stopped-flow spectroscopy, we established that the equilibrium between free VceR and VceR-CCCP plays a critical role in controlling expression of the pump. The co operative transition between these states allows the repressor to respond to relatively small changes in drug concentration. Thus, repression and induction can be readily switched about a critical drug concentration which will prove toxic to the cell. PMID- 15890204 TI - Kinetic analysis of the interactions of human papillomavirus E6 oncoproteins with the ubiquitin ligase E6AP using surface plasmon resonance. AB - Cervical cancers evolve from lesions generated by genital human papillomaviruses (HPV). "Low-risk" genital HPVs cause benign proliferations whereas "high-risk" types have the potential to progress into cancer. High-risk HPV E6 oncoproteins interact with the ubiquitin ligase E6AP and target several cellular proteins, including p53 and proteins of the MAGI family, towards ubiquitin-mediated degradation. E6AP, like other E6 binding proteins such as E6BP, IRF-3 and paxillin, interacts with E6 via a consensus leucine-charged motif. Here we have investigated the kinetics of the interactions of a 15-mer peptide containing the LxxvarphiLsh motif of E6AP with E6. For this we have developed a Biacore assay based on antibody-capture on the sensor surface of GST- and/or MBP-E6AP peptide constructs followed by E6 protein injection. Our experiments show that E6 oncoproteins from four major high-risk (16, 18, 33 and 58) HPV types bind to E6AP with equilibrium dissociation constants in the low micromolar range. The kinetic dissociation parameters of these interactions are remarkably similar. On the other hand, low-risk HPV 11 E6 does not interact with E6AP even at relatively high concentrations. We also show that the two zinc-binding domains of E6 are required for E6AP recognition. Finally, we have analysed the binding properties of site-directed mutants of the E6AP-derived peptide. We demonstrate the importance for binding of conserved aliphatic side-chains and the moderate role of the global negative charge of the peptide. This work provides the first quantitative data on an HPV E6-mediated interaction, which support the current models of E6AP-mediated degradation. PMID- 15890206 TI - Nucleation and the transition state of the SH3 domain. AB - We present a verified computational model of the SH3 domain transition state (TS) ensemble. This model was built for three separate SH3 domains using experimental phi-values as structural constraints in all-atom protein folding simulations. While averaging over all conformations incorrectly considers non-TS conformations as transition states, quantifying structures as pre-TS, TS, and post-TS by measurement of their transmission coefficient ("probability to fold", or p(fold)) allows for rigorous conclusions regarding the structure of the folding nucleus and a full mechanistic analysis of the folding process. Through analysis of the TS, we observe a highly polarized nucleus in which many residues are solvent exposed. Mechanistic analysis suggests the hydrophobic core forms largely after an early nucleation step. SH3 presents an ideal system for studying the nucleation-condensation mechanism and highlights the synergistic relationship between experiment and simulation in the study of protein folding. PMID- 15890205 TI - A single amino acid substitution in the active site of Escherichia coli aspartate transcarbamoylase prevents the allosteric transition. AB - Modeling of the tetrahedral intermediate within the active site of Escherichia coli aspartate transcarbamoylase revealed a specific interaction with the side chain of Gln137, an interaction not previously observed in the structure of the X ray enzyme in the presence of N-phosphonacetyl-L-aspartate (PALA). Previous site specific mutagenesis experiments showed that when Gln137 was replaced by alanine, the resulting mutant enzyme (Q137A) exhibited approximately 50-fold less activity than the wild-type enzyme, exhibited no homotropic cooperativity, and the binding of both carbamoyl phosphate and aspartate were extremely compromised. To elucidate the structural alterations in the mutant enzyme that might lead to such pronounced changes in kinetic and binding properties, the Q137A enzyme was studied by time-resolved, small-angle X-ray scattering and its structure was determined in the presence of PALA to 2.7 angstroms resolution. Time-resolved, small-angle X-ray scattering established that the natural substrates, carbamoyl phosphate and L-aspartate, do not induce in the Q137A enzyme the same conformational changes as observed for the wild-type enzyme, although the scattering pattern of the Q137A and wild-type enzymes in the presence of PALA were identical. The overall structure of the Q137A enzyme is similar to that of the R-state structure of wild-type enzyme with PALA bound. However, there are differences in the manner by which the Q137A enzyme coordinates PALA, especially in the side-chain positions of Arg105 and His134. The replacement of Gln137 by Ala also has a dramatic effect on the electrostatics of the active site. These data taken together suggest that the side-chain of Gln137 in the wild-type enzyme is required for the binding of carbamoyl phosphate in the proper orientation so as to induce conformational changes required for the creation of the high affinity aspartate-binding site. The inability of carbamoyl phosphate to create the high-affinity binding site in the Q137A enzyme results in an enzyme locked in the low-activity low-affinity T state. These results emphasize the absolute requirement of the binding of carbamoyl phosphate for the creation of the high affinity aspartate-binding site and for inducing the homotropic cooperativity in aspartate transcarbamoylase. PMID- 15890207 TI - Prediction of multimolecular assemblies by multiple docking. AB - The majority of proteins function when associated in multimolecular assemblies. Yet, prediction of the structures of multimolecular complexes has largely not been addressed, probably due to the magnitude of the combinatorial complexity of the problem. Docking applications have traditionally been used to predict pairwise interactions between molecules. We have developed an algorithm that extends the application of docking to multimolecular assemblies. We apply it to predict quaternary structures of both oligomers and multi-protein complexes. The algorithm predicted well a near-native arrangement of the input subunits for all cases in our data set, where the number of the subunits of the different target complexes varied from three to ten. In order to simulate a more realistic scenario, unbound cases were tested. In these cases the input conformations of the subunits are either unbound conformations of the subunits or a model obtained by a homology modeling technique. The successful predictions of the unbound cases, where the input conformations of the subunits are different from their conformations within the target complex, suggest that the algorithm is robust. We expect that this type of algorithm should be particularly useful to predict the structures of large macromolecular assemblies, which are difficult to solve by experimental structure determination. PMID- 15890208 TI - Benefits of intense glucose control in critically ill patients. PMID- 15890209 TI - Iatrogenic peripheral nerve injuries. PMID- 15890210 TI - Peritoneal carcinomatosis. PMID- 15890211 TI - Chronic rejection and its multifactorial causes. PMID- 15890212 TI - Living donor liver transplantation. PMID- 15890213 TI - Transplantation of the pancreas and pancreatic islets: has it lived up to its promise? PMID- 15890214 TI - An update on the surgical management of colon cancer. PMID- 15890215 TI - Laparoscopic colorectal surgery: indications and techniques. PMID- 15890216 TI - Diarrhea and weight loss in a recent emigre. PMID- 15890217 TI - Cameron Haight: pioneer in the treatment of esophageal atresia. PMID- 15890218 TI - Establishing learning curves for surgical residents using Cumulative Summation (CUSUM) Analysis. AB - BACKGROUND: The assessment of technical proficiency is of paramount importance in the training of surgical residents. The fact that technical proficiency is underrepresented in the context of the ACGME outcomes project is evidenced in that proficiency skills comprise less than 5% of all assessments that evaluate residents. In this study, we use Cumulative Summation Analysis (CUSUM) as a visual objective analytic tool to determine performance accuracy and establish learning curves for PGY-1s in surgery. METHODS: From April 2001 to May 2002, 11 surgical residents completed a 1-month anesthesia rotation. Each resident was asked to complete a preoperative airway assessment followed by endotracheal intubation with induction of anesthesia. Airway assessment was performed independently by a resident and a licensed anesthesiologist or certified anesthetist with the modified Mallampati Score. Data were sequentially collected and plotted for summated successes and failures. RESULTS: The average intern required approximately 19 intubation attempts to complete the learning curve experience. There was no learning curve for airway assessment. CONCLUSIONS: The CUSUM analysis is an effective objective tool to define learning curves for technical skills. Vital information is provided for surgical programs that place residents in positions to manage airways, and limitless potential for defining the learning curves for technical skills is provided. PMID- 15890219 TI - Extremely indirect dermal lymphatic channels: value of areolar-cutaneous junction injection technique for breast lymphoscintigraphy. AB - This case describes a patient who initially had peri-excisional injections of radiocolloid around a lumpectomy site that failed to visualize sentinel nodes during lymphoscintigraphy. After additional areolar-cutaneous junction injections, extremely indirect dermal lymphatic pathways were evident with resultant drainage to two sentinel nodes, both of which contained disease. Hybrid combination injections of radiotracer during sentinel lymph node biopsy is gaining in popularity. This case illustrates the validity of the hybrid injection techniques and the demonstrated accuracy despite seemingly disconnected and extremely tortuous pathways as demonstrated in the figure. PMID- 15890220 TI - Perforated solitary diverticulitis of the ascending colon. PMID- 15890221 TI - Primary pulmonary spindle cell neoplasm. PMID- 15890222 TI - Surgery resident learning styles and academic achievement. AB - OBJECTIVE: To determine if surgical residents share a preferred learning style as measured by Kolb's Learning Style Inventory (LSI) and if a relationship exists between resident learning style and achievement as measured by a standardized examination (AME). Also, core faculty learning styles were assessed to determine if faculty and residents share a preferred learning style. DESIGN: Kolb's LSI, Version 3, was administered to 16 surgical residents and the residency program's core faculty of 6 attending physicians. To measure academic achievement, the American Medical Education (AME) examination was administered to residents. SETTING: The Hospital of Saint Raphael, General Surgery Residency Program, New Haven, Connecticut. Both instruments were administered to residents during protected core curriculum time. Core faculty were administered the LSI on an individual basis. PARTICIPANTS: Surgical residents of the Hospital of Saint Raphael's General Surgery Residency Program and 6 core faculty members RESULTS: Analysis of resident learning style preference revealed Converging as the most commonly occurring style for residents (7) followed by Accommodating (5), Assimilating (3), and Diverging (1). The predominant learning style for core faculty was also Converging (4) with 2 Divergers. The average score for the Convergers on the AME was 62.6 compared with 42 for the next most frequently occurring learning style, Accommodators. CONCLUSIONS: In this surgical residency program, a preferred learning style for residents seems to exist (Converging), which confirms what previous studies have found. Additionally, residents with this learning style attained a higher average achievement score as measured by the AME. Also, core faculty share the same preferential learning style as this subset of residents. PMID- 15890223 TI - Transaxillary thoracoscopically assisted sympathectomy for nonhealing ulcers in scleroderma: a case report of successful treatment. AB - We describe the successful treatment of a patient with chronic nonhealing ulcers caused by scleroderma refractory to medical therapy. This patient underwent a successful, transaxillary, thoracoscopically assisted, limited sympathectomy that resulted in healing of her ulcers and resolution of pain in the affected hand. PMID- 15890224 TI - Code. PMID- 15890225 TI - A look at objective resident evaluation. PMID- 15890226 TI - A multifaceted approach to the evaluation of surgical residents: a resident's view. PMID- 15890227 TI - Quality improvement in surgical education through objective resident evaluation. PMID- 15890228 TI - Complications of low-molecular-weight heparin in surgical patients with impaired renal function. PMID- 15890229 TI - Laparoscopic resection of infarcted appendices epiploicae of the colon. PMID- 15890230 TI - Advanced optical imaging requiring no contrast agents--a new armamentarium for medicine and surgery. PMID- 15890231 TI - Laminin-induced signaling in tumor cells. AB - Laminin is the main non-collagenous glycoprotein found in the basement membrane. The various laminin isoforms are involved in many physiological and pathological processes, including cancer dissemination. The interaction of cancer cells with laminin was identified as a key event in tumor invasion and metastasis. Laminin effects are mediated by laminin receptors that are divided into two groups: integrin and non-integrin receptors. Activation of a specific signal transduction pathway in the cell depends on various factors and may be altered when normal tissue becomes neoplastic. Laminin signals via multiple signal transduction pathways involving various components such as G-proteins, intracellular calcium, phospholipase D, mitogen activated protein kinases, phosphatases, focal adhesion kinase, small GTPases of the Rho family, and cytoskeleton components. This review focuses on the role of laminin in tumor progression, its signaling via the non integrin 67kDa laminin receptor and via integrins and the reciprocal relations between these receptors in certain tumors. PMID- 15890232 TI - Expression of peroxisome proliferator-activated receptor delta in human gastric cancer and its response to specific COX-2 inhibitor. AB - Peroxisome proliferator-activated receptor delta (PPARdelta) is ligand-activated transcription factor of the nuclear receptor superfamily which is recently implicated in carcinogenesis. We examined the expression profiles of PPARdelta in human gastric cancer, normal gastric mucosa and gastric cancer cell lines by RT PCR, Western blot and immunohistochemistry. PPARdelta mRNA and protein was found to be ubiquitously expressed in all 5 gastric cancer cell lines, 40 gastric cancer samples and 10 normal gastric mucosa from non-cancer patients. Positive immunoreactivity was detected in the nuclei of normal and malignant gastric epithelium. Treatment of gastric cell line MKN45 that overexpressed cyclooxygenase-2 (COX-2) with specific COX-2 inhibitor NS398 resulted in a time- and dose-dependent suppression of PPARdelta expression. In contrast, there was no suppression of PPARdelta in MKN28 gastric cell line with low COX-2 expression. Our results demonstrated the ubiquitous expression of PPARdelta in normal and cancer gastric epithelium. Suppression of PPARdelta may be one of the mechanisms underlying the chemopreventive effects of COX-2 inhibitor. PMID- 15890233 TI - Cytotoxicity of phenolic acid phenethyl esters on oral cancer cells. AB - Many phenolic acid phenethyl esters possess diverse biological effects including anti-cancer activity. A series of 14 derivatives were synthesized for the evaluation of their cytotoxic effect on oral cancer cells. These derivatives were tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric and trypan blue dye exclusion assay on the growth of oral squamous cell carcinoma (SAS), oral epidermoid carcinoma-Meng 1 (OEC-M1), and normal human oral fibroblast (NHOF) cells, respectively. Caffeic acid phenethyl esters, 3a (CAPE), and 3b, 3c, and 3d showed cytotoxic effects on the SAS and OEC-M1 cell lines, but not the NHOF cell line at a 5-100 microM dose range. Flow cytometric analysis showed that 3c caused OEC-M1 cell arrest at G2/M phase. Such differential effects on representative cancer and normal cells suggested these compounds might be useful in oral cancer chemotherapy. PMID- 15890234 TI - Mouse skin tumor promotion by sodium arsenate is associated with enhanced PCNA expression. AB - Drinking water contamination by arsenicals remains a major public health problem in many parts of the world more particularly in India and Bangladesh. Despite arsenic being a health hazard and implicated in human carcinogenesis, the experimental evidence available is much limited even now and the mechanisms involved during carcinogenesis and tumor promotions are not clear. Accordingly, in this study, we have studied the tumor promoter effects of sodium arsenate on mouse skin tumor promoter model system using 9,10-dimethyl-1,2-benzanthracene (DMBA) as a initiating carcinogen. Our studies showed development of papillomas on mice skin treated with only DMBA. However, mice treated with DMBA on skin and administered arsenate (As) in drinking water showed development of well differentiated squamous cell carcinomas. Further, both by immunohistochemistry and western blotting analysis studies higher levels of proliferating cell nuclear antigen (PCNA) was observed in mice treated with DMBA plus arsenate compared to only DMBA treated group. PCNA is known to be associated with S phase and DNA replication of the cell cycle. The plain controls and arsenate controls did not show significant difference either in tumor development or in PCNA levels. The present study demonstrates mouse skin tumor promoting effect of arsenate which seems to be associated with abnormal cell proliferation as indicated by higher levels of PCNA expression. PMID- 15890235 TI - Lack of prepubertal administration of ethinyl estradiol on susceptibility to multiple organ carcinogenesis in rats exposed to 7,12-dimethylbenz[a]anthracene and N-bis(2-hydroxypropyl)nitrosamine during adolescence. AB - Estrogen exposure during the adult period is widely known to promote tumor development in the female genital system, as well as in the mammary gland in experimental animals, but its carcinogenic potential with exposure at the prepubertal stage, for 6 weeks after birth, is not completely understood. In the present study, we therefore evaluated the modifying effects of prepubertal ethinyl estradiol (EE) treatment on susceptibility to multiple organ carcinogenesis with subsequent carcinogen exposure in F344 rats. Dams during the lactation period and their weaned offspring until postnatal-week 6 were fed diet containing 0, 0.2 or 1.0 ppm EE. The offsprings were then administered 7,12 dimethylbenz[a]anthracene (DMBA, 50mg/kg body weight) by gavage for mammary tumor induction in week 7 and given free access to drinking water containing N-bis (2 hydroxypropyl)nitrosamine (DHPN, 0.2%) for wide spectrum tumor induction in organs such as the thyroid, liver, lung and kidney from weeks 6-14. Male and female offspring were euthanized at weeks 27 and 36, respectively, for histopathological examination. While the incidence and multiplicity of mammary tumors showed a tendency for increase in females of the 0.2 and 1.0 ppm EE groups, this was without statistical significance. Furthermore, prepubertal EE exposure did not affect tumor induction in the thyroid, liver, lung, kidney, esophagus, ovary and lymphoid tissue in either sex. The present results thus indicate a lack of influence of estrogen early in life on carcinogenic susceptibility, although the possible impact on mammary carcinogenesis requires further examination. PMID- 15890236 TI - Growth inhibitory effect of alk(en)yl thiosulfates derived from onion and garlic in human immortalized and tumor cell lines. AB - Two alk(en)yl thiosulfates, sodium n-propyl thiosulfate (NPTS) and sodium 2 propenyl thiosulfate (2PTS), are natural constituents of onion and garlic, respectively, which were identified originally as causative agents of onion- and garlic-induced hemolytic anemia in dogs. As a continuation of our studies on the beneficial functions of NPTS and 2PTS, in the present study, we investigated the antitumor effects of these compounds. They were shown to inhibit the in vitro proliferation of three human tumorigenic cell lines, WiDr, 293 and HL-60, in a dose-dependent manner. Overall, NPTS seemed to have weak activity for inhibiting cell growth compared with 2PTS, though not in WiDr cells, which were sensitive to both compounds. NPTS and 2PTS caused oxidative damage to HL-60 cells and induced apoptosis. The extent of apoptosis was approximately proportional to that of the oxidative damage and also to that of the cytotoxicity caused by these compounds. These results suggest that the alk(en)yl thiosulfates have an antitumor effect through the induction of apoptosis initiated by oxidative stress. PMID- 15890237 TI - Cytotoxicity determination without photochemical artifacts. AB - A study was performed to improve cytotoxicity determinations by eliminating flavin-mediated photosensitization from tests with KB cells, NCI-H69 cells, P glycoprotein expressing KBC5-8 cells, MRP1-expressing H69AR cells, and A240286S human lung adenocarcinoma cells. Growth inhibition by cis-platin, doxorubicin, etoposide, gemcitabine, taxol, vincristine, vinblastine, and vinorelbine was determined under flavin-protecting conditions using flavin-free culture media with fetal bovine serum as the only source of flavins. As compared to conventional tests, the IC50 values determined under flavin-protecting conditions reflected increased apparent drug cytotoxicities, and were flawlessly reproducible. Flavin-mediated photosensitization should, therefore, be strictly eliminated from in vitro experiments involving cytotoxic and other drugs. PMID- 15890238 TI - Silencing of the p53R2 gene by RNA interference inhibits growth and enhances 5 fluorouracil sensitivity of oral cancer cells. AB - The p53R2 gene encodes the ribonucleotide reductase (RR) small subunit 2 homologue, and is induced by several stress signals activating p53, such as DNA damaging agents. The p53R2 gene product causes an increase in the deoxynucleotide triphosphate (dNTP) pool in the nucleus, which facilitates DNA repair and synthesis. We hypothesized that p53R2 would be a good molecular target for cancer gene therapy. In this study, three human oral cancer cell lines (SAS, HSC-4 and Ca9-22), a human breast cancer cell line MCF-7, and a normal human fibroblast cell line NHDF were tested. We silenced the expression of p53R2 with the highly specific post-transcriptional suppression of RNA interference (RNAi). We investigated p53R2 expression with the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The sensitivity to anticancer agents was evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of p53R2 showed no association with the mutational status of p53. The cancer cell lines with higher p53R2 expression were more resistant to 5-FU. RNAi-mediated p53R2 reduction selectivity inhibited growth and enhanced chemosensitivity in cancer cell lines but not in normal fibroblasts. These results suggest that basal transcription of p53R2 could be associated with the sensitivity to anticancer agents. Moreover, we assessed the possibility that p53R2 would be a good molecular target, and report that RNAi targeting of p53R2 could be useful for oral cancer gene therapy. PMID- 15890239 TI - Regulation of activity of cathepsins B, L, and D in murine lymphosarcoma model at a combined treatment with cyclophosphamide and yeast polysaccharide. AB - Changes in the activity of cysteine (cathepsins B and L) and aspartyl (cathepsin D) proteases were investigated at the development of susceptible and resistant variants of murine lymphosarcoma (LS). It has been demonstrated that the variant resistant to the cyclophosphamide treatment is characterized by a lower activity of all three cathepsins in the tumor tissue. Application of a higher dose of cyclophosphamide led to a more pronounced increase of the studied enzymatic activity in mice with a resistant variant of LS, than in those with a susceptible one. Administration of a yeast polysaccharide derivative - sulfoethyl glucan - enhanced therapeutic effect of cyclophosphamide in mice with both variants of LS, while the most efficient dose was found to be that of 10mg/kg body mass. In the intact mice, usage of both cyclophosphamide and sulfoethyl glucan led to a similar increase of the cathepsins activity in liver and spleen. PMID- 15890240 TI - Associated nutritional risk of breast and colon cancers: a population-based case control study in Montreal, Canada. AB - To investigate whether particular patterns of food consumption are related to the occurrence of breast and colon cancer, two simultaneous case-control studies of French-Canadians with 616 cases (414 breast cancer, 202 colon cancer) and 429 population-based controls were carried out. Food consumption data were obtained via a validated food frequency questionnaire. Three dietary patterns were identified. The 'chocolate-cereal' pattern was typified by a high intake of chocolate-based products, breakfast cereals, water, and fruits. The 'pork and processed meat' pattern was characterized by a high intake of pork, white bread and processed meat products, with the 'drinker' pattern depicted by elevated consumption of wine, liquor, and beer. After adjustment for determinants of breast and colon cancers, no significant association was apparent between the three dietary patterns and breast or colon cancer. These results suggest that the observed breast and colon cancer relationship may not be a result of food patterns in French-Canadian women. PMID- 15890241 TI - NAT2 fast acetylator genotype is associated with an increased risk of lung cancer among never-smoking women in Taiwan. AB - The correlation between cooking oil fumes, containing relatively higher amounts of heterocyclic amines, and female lung cancer has been revealed. The association of genetic polymorphisms of CYP1A2 and NAT2, two major enzymes responsible for the metabolism of heterocyclic amines, with lung cancer has been investigated with inconclusive results. In this study targeted on never-smoking population with 162 lung cancer patients and 208 non-cancer controls, while the distributions of CYP1A2 phenotypes in lung cancer patients were comparable to that in controls, NAT2 fast acetylators had an OR of 2.44 (95% CI 1.40-4.23, P=0.002) and 2.56 (95% CI 1.37-4.80, P=0.003) for lung cancer in overall and female cases, respectively, but not in males. These results suggested never smoking females with NAT2 fast acetylator were more prone to lung cancer and reflected the possibility that exposure to heterocyclic amines may contribute to the female lung cancer development in Taiwan. PMID- 15890242 TI - Thymidylate synthase and dihydropyrimidine dehydrogenase gene expression in breast cancer predicts 5-FU sensitivity by a histocultural drug sensitivity test. AB - Thymidylate synthase (TS), Dihydropyrimidine dehydrogenase (DPD) and Thymidine Phosphorylase (TP) gene expressions are reported to be predictive markers for 5 fluorouracil (5-FU) sensitivity in gastrointestinal cancer. However, in breast cancer, it is still controversial whether those molecular markers predict 5-FU sensitivity or not. One possible reason for the difficulty may be the histological heterogeneity in breast cancer specimens. In this study, TS, DPD and TP mRNA expression in 40 breast cancer tumors were semi-quantified separately in cancer cells (Ca), cancerous stroma (Str) and normal glands (Nor) using laser capture microdissection and real time RT-PCR (LCM+RT-PCR). The histoculture drug response assay (HDRA) for 5-FU sensitivity was performed for 22 tumors. TS and TP mRNA expressions were higher in Ca than Str, although DPD gene expression was lower in Ca than Str. The group of high TS and high DPD gene expression in Ca was resistant to 5-FU, and the group of low TS and low DPD gene expression in Ca was sensitive to 5-FU (P=0.048 chi-square test). TS and DPD mRNA expressions measured using LCM+RT-PCR might be useful predictive markers for 5-FU sensitivity in human breast cancer. PMID- 15890243 TI - Polymorphism in the genes of alpha and beta tumor necrosis factors (TNF-alpha and TNF-beta) and gamma interferon (IFN-gamma) among Iranian women with breast cancer. AB - The association of the polymorphism in the genes of TNF-alpha, TNF-beta and IFN gamma with breast cancer was investigated in the present study. There were no differences in the TNF-alpha and TNF-beta alleles and genotypes frequencies between breast cancer patients (n=223) and control subjects (n=267). The correlations between TNFA or TNFB alleles or genotypes and clinicopathological indices were also insignificant. However, the frequency of IFN-gamma+874 T/T genotype was significantly higher in breast cancer patients compared to those of controls (P<0.002; OR=2.03, 95% CI=1.28-3.2) which indicates that Iranian women carrying the IFN-gamma+874 T/T genotype may be exposed to an increased risk of breast cancer development. PMID- 15890244 TI - Prostate cancer mediates osteoclastogenesis through two different pathways. AB - The present study was undertaken to test the effects of prostate cancer cell lines (LNCaP, DU145, PC3, and MDA PCa 2b) on osteoclastogenesis. Crude conditioned medium (CM) from all four prostate cancer cell lines enhanced expression of the mRNA for receptor activator of NF-kappaB ligand (RANKL) in a mouse osteoblast cell line, MC3T3-E1; however, CM had no effect on expression of osteoprotegerin (OPG) mRNA. Coculture of MC3T3-E1 with prostate cancer cells yielded similar results. The number of mature osteoclasts induced by soluble RANKL increased significantly when osteoclast precursor cells were cultured with CM from LNCaP and DU145 cells. CM from LNCaP and DU145 cells also induced maturation from precursor in the absence of soluble RANKL, and this effect was not blocked by OPG. Addition of CM from DU145 cells increased expression of MMP-9 mRNA by osteoclast precursors. Our findings indicate that prostate cancer mediates osteoclastogenesis through induction of RANKL expression by osteoblasts and through direct actions on osteoclast precursors mediated by some factors other than RANKL. PMID- 15890245 TI - Gastric cancer cell detection in peritoneal lavage: RT-PCR for carcinoembryonic antigen transcripts versus the combined cytology with peritoneal carcinoembryonic antigen levels. AB - Free cancer cells exfoliated from the serosal surfaces of primary cancers are considered to be responsible for peritoneal dissemination, which are both the most frequent pattern of treatment failure and the most important cause of death in gastric cancer patients. Detection of free cancer cells in the peritoneal cavity at the time of surgery, therefore, is thought to be of great value in predicting peritoneal recurrence and accordingly the prognosis of gastric cancer patients. This study was designed to determine whether free cancer cells in peritoneal lavage fluid from gastric cancer patients could be identified by a reverse-transcriptase polymerase chain reaction (RT-PCR) method specific to carcinoembryonic antigen (CEA) mRNA. Simultaneously, the results from conventional cytological examination were evaluated and the levels of CEA in peritoneal lavage were determined. Of the 40 gastric cancer patients enrolling in this investigation, 11 (27.5%) were positive for CEA mRNA in their peritoneal lavage, whereas only 6 (15%) and 8 (20%) were shown to be positive by cytological examination and peritoneal CEA (pCEA) assay, respectively. Furthermore, RT-PCR positive for CEA mRNA was correlated with the depth of tumor invasion (P<0.001), lymph node metastases (P=0.004), the TNM stage (P<0.001) and peritoneal recurrence (P<0.001). The technique of RT-PCR was more sensitive than conventional cytological examination and pCEA levels in the detection of peritoneal free cancer cells as well as in the prediction of peritoneal recurrence. In addition, CEA RT-PCR had a high concordance rate (82.5%) with the combination of cytology with pCEA levels. These observations suggest that it is feasible to identify free cancer cells in peritoneal lavage by using a CEA mRNA specific RT-PCR method, and this assay can be a promising diagnostic modality for evaluating the risk of peritoneal dissemination in gastric cancer patients following operations. PMID- 15890246 TI - Alterations in the tissue inhibitor of metalloproteinase-3 (TIMP-3) are found frequently in human colorectal tumours displaying either microsatellite stability (MSS) or instability (MSI). AB - Methylation of promoter regions and frameshift mutations in microsatellites of the coding sequence (CDS) of genes are frequently associated with loss of expression in microsatellite instable (MSI) colorectal carcinoma. In a panel of 40 MSI and 24 microsatellite stable (MSS) colorectal tumours as well as six cultured colorectal carcinoma cell lines hypermethylation of the TIMP3-promoter was found in 28% of MSI and 25% of MSS tumours, respectively. Additionally, three MSI tumours and one cell line displayed instability of a C7-repeat located in the CDS of the TIMP-3 gene. TIMP-3 fulfils all important criteria for being a target gene in the mutator pathway. Thus, TIMP-3 might be a factor of general importance for colorectal carcinogenesis. PMID- 15890247 TI - Roles of mismatch repair proteins hMSH2 and hMLH1 in the development of sporadic breast cancer. AB - Defects in mismatch repair (MMR) genes, particularly the hMSH2 and hMLH1 genes, are associated with a variety of cancers including sporadic breast cancer. However, whether or not patient clinical background, e.g. age, progesterone receptor (PR), estrogen receptor (ER), tumor progression and stage, chemotherapy history, and menopausal status, influences MMR status is not understood. To address these issues, 83 archival breast cancer specimens were examined for expression of hMSH2 and hMLH1 by immunohistochemistry and the relationship between MMR protein expression and patient clinical background was analyzed. We detected lack of or reduced expression of hMSH2 and hMLH1 in 23 (27.7%) and 26 cases (31.3%), respectively, and hypermethylation of the hMLH1 promoter accounted for the majority of the cases with reduced expression of hMLH1. Statistical analysis revealed that (i) reduced expression of hMLH1 and hMSH2 seemed to confer advantage for the progression of breast tumors to more advanced stages; (ii) attenuated expression of hMLH1 correlated with history of chemotherapy, but not with age, menopause, or the status of PR and ER; (iii) hypermethylation of the hMLH1 promoter was linked with clinical stage and lymphatic metastasis. These analyses indicate that defective expression of MMR genes is closely associated with the development of sporadic breast cancer. PMID- 15890248 TI - Inhibition of lipoxygenase pathway in macrophages co-cultivated with tumor cells. AB - Although there is a great deal of interest in the role played by tumor-associated macrophages in tumor progression, the knowledge of the biological mediators involved in the interplay between macrophages and tumor cells is still limited. In the present study, we investigated whether the lipoxygenase pathway in resident murine peritoneal macrophages is affected by contact with tumor cells of a different origin, e.g. murine B16 melanoma and L929 fibrosarcoma cells, and human Hs294T melanoma and HT1080 fibrosarcoma cells. Our experiments have been carried out by using macrophages co-cultivated with tumor cells at different ratios, in order to simulate the relative proportions between macrophages and tumor cells during the in vivo development of a tumor. Reverse phase HPLC analyses of the lipoxygenase products of resident peritoneal macrophages revealed a rather complex profile characterized by a high level of 12(S) hydroxyeicosatetraenoic acid and 15(S)-hydroxyeicosatetraenoic acid followed by leukotriene B(4), 5(S)-hydroxyeicosatetraenoic acid, and lipoxins. Macrophages co cultivated with tumor cells, both murine and human, showed a marked reduction of lipoxygenase products, mainly in the co-cultures where tumor cells prevailed over macrophages. The characteristic profile of macrophage lipoxygenase products was re-established after removal of tumor cells from the co-cultures. The inhibitory effect on lipoxygenase pathways exerted by tumor cells, was not seen when macrophages were co-cultivated with normal primary murine and human fibroblasts. PMID- 15890249 TI - Forced expression of deltaN-TCF-1B in colon cancer derived cell lines is accompanied by the induction of CEACAM5/6 and mesothelin. AB - The colon cancer cell lines HT29 and SW480 were transfected with an N-terminal beta-catenin binding site-deficient high mobility group (HMG)-box T-cell factor 1 (deltaN-TCF-1) construct to identify differentially expressed genes. Oligonucleotide HG-U133A microarray expression profiling revealed increased mRNA levels of carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 5, 6 and mesothelin in transfectants positive for nuclear deltaN-TCF-1B. Increased amounts of CEACAM5 (CEA) were detectable in membrane-associated compartments, particularly in cholesterol-enriched microdomains. Similarly, mesothelin was demonstrated as an uncleaved membrane-bound constituent. The identified markers were examined in specimens of 46 colorectal carcinomas (CRC) by immunohistochemistry. Patchy areas of increased CEACAM5/6 staining were seen at the tumour-host front in all samples studied. Twenty-eight (58%) of these cases showed over-expression of mesothelin in a small fraction of tumor cells displaying dedifferentiation and dissemination at the invasion front. We conclude that forced expression of deltaN-TCF-1B in HT29 and SW480 is associated with up regulation of GPI-anchored adhesion molecules, which were assigned to the tumour host front in CRC patients. PMID- 15890250 TI - Chromosomal imbalances in sporadic neuroendocrine tumours of the thymus. AB - Neuroendocrine (carcinoid) tumours of the thymus are rare neoplasms characterized by a highly malignant clinical behavior. Some of these tumors are associated with MEN1. In this study we evaluated 10 cases of sporadic thymic neuroendocrine tumours using immunohistochemistry and comparative genomic hybridization (CGH). All tumours showed a diffuse expression of neuron specific enolase (NSE) and synaptophysin. Chromosomal imbalances were detected in 8/10 cases, the most frequent gains were seen on chromosome Xp (3/10 cases), 7p, 7q, 11q, 12q, and 20q (2/10 each), losses were most frequently detected at 6q (5/10 each), 6p (3/10 each), 4q (3/10 each), 3p, 10q, 11q and 13 q (2/10 each). These CGH data show a degree of overlap with chromosomal imbalances commonly observed in advanced thymomas. PMID- 15890251 TI - Rigid overlay of volume sonography and MR image data of the female pelvic floor using a fiducial based alignment--feasibility due to a case series. AB - The visual combination of different medical image acquisition techniques (modalities) can lead to new modalities with enhanced informative content. In this paper, we present an overlay technique of magnetic resonance (MR) and 3D US image data sets of the female anal canal (internal and external sphincter) as a base for a new diagnostic modality. It is a new field of the application of the overlay technique. Three corresponding MR and US volume data sets from the female pelvic floor region were filtered using adaptive filtering techniques and overlayed (=registered rigidly) with a landmark based alignment method. PMID- 15890252 TI - Fast colon centreline calculation using optimised 3D topological thinning. AB - Topological thinning can be used to accurately identify the central path through a computer model of the colon generated using computed tomography colonography. The central path can subsequently be used to simplify the task of navigation within the colon model. Unfortunately standard topological thinning is an extremely inefficient process. We present an optimised version of topological thinning that significantly improves the performance of centreline calculation without compromising the accuracy of the result. This is achieved by using lookup tables to reduce the computational burden associated with the thinning process. PMID- 15890253 TI - Evaluation of osteoporosis in X-ray CT examination: a preliminary study for an automatic recognition algorithm for the central part of a vertebral body using abdominal X-ray CT images. AB - We have developed an algorithm that can distinguish the central part of the vertebral body from abdominal X-ray CT images to determine whether it is possible to aid a diagnosis of osteoporosis. We classified three measures for the principal component analysis and linear discriminant function. When we judged whether patients had osteoporosis or not, the ratio usable for diagnosing osteoporosis (sensitivity) was 1.00 (15/15), and for diagnosing as normal (specificity) was 0.64 (7/11). We believe that this algorithm can be used to aid in diagnosing osteoporosis, utilizing the measures obtained from the CT images. PMID- 15890254 TI - MRI diffusion-based filtering: a note on performance characterisation. AB - Frequently MRI data is characterised by a relatively low signal to noise ratio (SNR) or contrast to noise ratio (CNR). When developing automated Computer Assisted Diagnostic (CAD) techniques the errors introduced by the image noise are not acceptable. Thus, to limit these errors, a solution is to filter the data in order to increase the SNR. More importantly, the image filtering technique should be able to reduce the level of noise, but not at the expense of feature preservation. In this paper we detail the implementation of a number of 3D diffusion-based filtering techniques and we analyse their performance when they are applied to a large collection of MR datasets of varying type and quality. PMID- 15890255 TI - Fluid-attenuated inversion-recovery MR imaging in assessment of intracranial oligodendrogliomas. AB - This retrospective study consisted of 17 consecutive patients with oligodendrogliomas. We qualitatively and quantitatively assessed the diagnostic value of fluid-attenuated inversion-recovery (FLAIR) images compared with T2 weighted fast spin-echo (FSE) images for evaluating intracranial oligodendrogliomas. Qualitative evaluations of signal intensity, tumor conspicuity, definition of tumor margin, distinction between solid and cystic like parts within tumor, and calcification were performed. Quantitative criteria comparing FLAIR to T2-weighted FSE images included tumor-to-background contrast and contrast-to-noise ratio (CNR) and tumor-to-cerebrospinal fluid (CSF) contrast and CNR. Our results demonstrate that the FLAIR sequence can replace the T2 weighted FSE sequence for evaluating oligodendrogliomas. PMID- 15890256 TI - Multi-spectral image analysis and classification of melanoma using fuzzy membership based partitions. AB - The sensitivity and specificity of melanoma diagnosis can be improved by adding the lesion depth and structure information obtained from the multi-spectral, trans-illumination images to the surface characteristic information obtained from the epi-illumination images. Wavelet transform based bi-modal channel energy features obtained from the images are used in the analysis. Methods using both crisp and fuzzy membership based partitioning of the feature space are evaluated. For this purpose, the ADWAT classification method that uses crisp partitioning is extended to handle multi-spectral image data. Also, multi-dimensional fuzzy membership functions with Gaussian and Bell profiles are proposed for classification. Results show that the fuzzy membership functions with Bell profile are more effective than the extended ADWAT method in discriminating melanoma from dysplastic nevus. PMID- 15890257 TI - Development of wavelet de-noising technique for PET images. AB - Positron emission tomography (PET) imaging provides the functional information and precise physiological uptake of radioactivity in a patient's body. But the shortcoming of PET is low signal to noise ratio (SNR) due to photon noise. The noise may influence image quality, and cause the mistake of clinical interpretation. The purpose of this research is to develop a wavelet de-noising technique to reduce the noise of PET images. By processing the image through the optimum wavelet parameters we selected, we keep the resolution and contrast but reduce almost half of coefficient of variation in the region of interest of PET images. PMID- 15890258 TI - Progressive lossless compression of volumetric data using small memory load. AB - Nowadays, applications dealing with volumetric datasets, Medical applications being a typical representative, have become possible even on low cost computers due to a rapid increase of computer memory and processing power. However, even today, dealing with volumetric datasets creates two considerable problems: slow visualization and large file sizes. While recently, due to significant progress in graphics hardware, real-time or near real-time volume visualization has become possible, volume compression still remains a problematic issue. This paper introduces a new method for lossless compression of volumetric datasets. It is based on quadtree encoding. The method consists of three steps: during initialization, so-called division quadtree is built. The smallest unit of the division quadtree is called basic macro-block. During the processing phase, Boolean intersection is built on pairs of quadtrees, and the differences are stored. In the last phase, the variable length encoding is applied to reduce the entropy among the differences. Proposed method supports progressive visualization, what is especially important when a transfer trough the internet is needed. To test the efficiency of this method it was compared to popular octree encoding scheme. The results proved that data coherence is exploited more sufficiently using proposed quadtree approach. Additional advantage of this approach is that the algorithm does not need a lot of memory space. Only two quadtrees of two consecutive slices need be loaded in the memory at the same time. This feature makes this algorithm extremely attractive for possible hardware implementation. This paper introduces a new method for the compression of volumetric datasets. It is based on quadtree encoding. This method consists of three steps: during initialization, a so-called division quadtree is built. The smallest, unit of the division quadtree is called a basic macro-block. A Boolean intersection is built on pairs of quadtrees during the processing phase and the differences are stored. In the last phase, variable length encoding is applied to reduce entropy among the differences. This method has been compared with the popular octree-based method and gives, in general, better compression results. In addition, this method can be realized using small on-board memory. PMID- 15890260 TI - Integrin evolution: insights from ascidian and teleost fish genomes. AB - Integrins are a family of alphabeta heterodimeric receptors essential to cell adhesion in all metazoans. In humans, the family consists of 18 alpha and 8 beta subunits that combine to form 24 dimers. Here, we present phylogenetic reconstructions for the alpha and beta integrin subunits based on sequences from 24 invertebrate and vertebrate species, including the fully sequenced genomes of the ascidian Ciona intestinalis (a urochordate) and the pufferfish Takifugu rubripes (a teleost). Both genomes contain integrin alpha subunits that have the inserted alphaI domain. As for the one alphaI domain containing integrin alpha subunit discovered earlier from the ascidian Halocynthia roretzi, the Ciona alphaI domains are missing the distinctive characteristics of mammalian collagen receptors and segregate from all vertebrate alphaI domain integrins in a phylogenetic tree, forming a new subgroup of alpha subunits with alphaI domains. Each of the pufferfish alphaI domain sequences does have characteristics of the collagen receptor alphaI domains, but no leukocyte-specific alphaI domains were found in pufferfish. Comparative protein modeling suggests that several of these fish alphaI domains are structurally compatible with binding to a GFOGER sequence in a collagen triple helix. PMID- 15890261 TI - Dissection of human tropoelastin: supramolecular organization of polypeptide sequences coded by particular exons. AB - Polypeptide sequences encoded by some exons of the human tropoelastin gene (EDP, elastin-derived peptide) have been analysed for their ability to coacervate and to self-assembly. The great majority of them were shown to form organized structures, but only a few were indeed able to coacervate. Negative staining and rotary shadowing transmission electron microscopy showed the polypeptides to adopt a variety of supramolecular organization, from filaments, as those typical of tropoelastin, to amyloid-like fibers. The results obtained gave significant insight to the possible roles played by specific polypeptide sequences of tropoelastin. PMID- 15890262 TI - Endogenous thrombospondin-1 is not necessary for proliferation but is permissive for vascular smooth muscle cell responses to platelet-derived growth factor. AB - We have reexamined the role of endogenous thrombospondin-1 (TSP1) in growth and motility of vascular smooth muscle cells (SMCs). Based on the ability of aortic derived SMCs isolated from TSP1 null mice and grown in the absence of exogenous TSP1 to grow at comparable rates and to a slightly higher density than equivalent cells from wild-type mice, TSP1 is not necessary for their growth. Low concentrations of exogenous TSP1 stimulate growth of TSP1 null SMCs, but higher doses of TSP1 or its C-terminal domain are inhibitory. However, SMCs from TSP1 null mice are selectively deficient in chemotactic and proliferative responses to platelet-derived growth factor and in outgrowth in three-dimensional cultures. Recombinant portions of the N- and C-terminal domains of TSP1 stimulate SMC chemotaxis through different integrin receptors. Based on these data, the relative deficiency in SMC outgrowth during an ex vivo angiogenic response of muscle tissue from TSP1 null mice is probably due to restriction of platelet derived growth factor dependent SMC migration and/or proliferation. PMID- 15890263 TI - Expression pattern of matrilins and other extracellular matrix proteins characterize distinct stages of cell differentiation during antler development. AB - Deer antler regeneration is a uniquely intense and complex process, which involves chondrogenic and intramembranous ossification. Cell differentiation in the developing antler of red deer, Cervus elaphus, was characterized with extracellular matrix markers. Expression of the four matrilin genes was monitored by immunohistochemistry and in situ hybridization and compared to cartilage markers collagen II and cartilage link protein, the bone component collagen I, and the endothelial basement membrane constituent laminin. The mesenchyme layer at the very tip of the velvet antler was enriched in link protein, indicative of the role of hyaluronan in apical morphogenesis. Matrilin-2, formerly described as a component of hard and soft connective tissue matrices, was identified here also as a marker of cells with high differentiation potential: it is expressed predominantly by mesenchyme cells, prechondrocytes and preosteoblasts. In addition to matrilin-3, documented as a component of the bony extracellular matrix, expression of the other three matrilin genes was observed in osteoprogenitor cells and osteoblasts. A layer of presumed osteoprogenitor cells, which surrounded the perivascular channels, expressed all four matrilins and collagen I. As a consequence, all four matrilins, including matrilin-1, previously detected in the skeleton only in cartilage, were found associated to collagen I-rich structures in a thin layer bordering the columns of hypertrophic chondrocytes. Cells with similar morphology and expression pattern were identified in the periosteum. Altogether all cell types of the chondrogenic and osteogenic lineage that expressed the four matrilins were in a separate study [Faucheux, C., Nicholls, B.M., Allen, S., Danks, J.A, Horton, M.A., Price, J.S., 2004. Recapitulation of the parathyroid hormone-related peptide-Indian hedgehog pathway in the regenerating deer antler. Dev. Dyn. 231, 88-97] positive for parathyroid hormone-related peptide and its receptor. PMID- 15890264 TI - Synthesis of prolyl 4-hydroxylase alpha subunit and type IV collagen in hemocytic granular cells of silkworm, Bombyx mori: Involvement of type IV collagen in self defense reaction and metamorphosis. AB - The present study shows that hemocytic granular cells synthesize and secrete type IV collagen (ColIV) in the silkworm Bombyx mori (B. mori) and suggests that these cells play roles in the formation of basement membrane, the encapsulation of foreign bodies, and the metamorphic remodeling of the gut. The full- and partial length cDNA of B. mori prolyl 4-hydroxylase alpha subunit (BmP4Halpha) and B. mori ColIV (BmColIV) were cloned, respectively. In situ hybridization and immunocytochemistry on larval tissues and cells identified hemocytic granular cells as the cells that express mRNAs and proteins of both BmP4Halpha and BmColIV. Immunohistochemistry and immunocytochemistry demonstrated that BmColIV was present in the basement membrane and in the secretory granules of granular cells, respectively. Granular cells in culture secreted BmColIV without accompanying the degranulation and discharged it from the granules when the cells were degranulated. Nylon threads were inserted into the hemocoel of larvae. Granular cells concentrated around the nylon threads and encapsulated them as a self-defense reaction. BmColIV was found to be a component of the capsules. Furthermore, the present study showed that actively BmColIV-expressing granular cells accumulated around the midgut epithelium and formed BmColIV-rich thick basal lamina-like structures there in larval to pupal metamorphosis. PMID- 15890265 TI - Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth. AB - The dental follicle is an ectomesenchymal tissue surrounding the developing tooth germ. It is believed that this tissue contains stem cells and lineage committed progenitor cells or precursor cells (PCs) for cementoblasts, periodontal ligament cells, and osteoblasts. In this study, we report the isolation of PCs derived from dental follicle of human third molar teeth. These fibroblast-like, colony forming and plastic adherent cells expressed putative stem cell markers Notch-1 and Nestin. We compared gene expressions of PCs, human mesenchymal stem cells (hMSCs), periodontal ligament cells (PDL-cells) and osteoblasts (MG63) for delimitation of PCs. Interestingly, PCs expressed higher amounts of insulin-like growth factor-2 (IGF-2) transcripts than hMSCs. Differentiation capacity was demonstrated under in vitro conditions for PCs. Long-term cultures with dexamethasone produced compact calcified nodules or appeared as plain membrane structures of different dimensions consisting of a connective tissue like matrix encapsulated by a mesothelium-like cellular structure. PCs differentially express osteocalcin (OCN) and bone sialoprotein (BS) after transplantation in immunocompromised mice but without any sign of cementum or bone formation. Therefore, our results demonstrate that cultured PCs are unique undifferentiated lineage committed cells residing in the periodontium prior or during tooth eruption. PMID- 15890266 TI - Hyaluronan content of Wharton's jelly in healthy and Down syndrome fetuses. AB - The mechanisms by which the excess genetic material of chromosome 21 results in the dysmorphologic features of Down syndrome (DS) are largely unknown. It has been found that the extracellular matrix of nuchal skin of DS fetuses exhibits an higher content of hyaluronan (HA) compared to that of euploid fetuses. Since HA plays a central role in many morphogenetic processes during embryogenesis, an alteration in its metabolism could be involved in the pathogenesis of several structural defects of DS. The extracellular matrix of umbilical cord (UC) is the mammalian tissue with one of the highest content of HA. Therefore we sought to explore the quantitative HA modifications during gestation, tissue distribution and HA metabolism in euploid and DS UCs. Euploid UCs (n=28) and UCs from DS fetuses (n=13) were obtained after termination of pregnancy, spontaneous abortion, or at delivery. Quantitative and molecular size analysis were performed using HPLC and FPLC. Tissue distribution was visualized by immunohistochemistry. Gene expression for HA synthases (HAS) and hyaluronidases (HYAL) were quantified by real-time PCR techniques and HYAL activity was detected by zymography. In euploid UC only HA of a molecular weight of 1700 kDA was present while in DS UC an additional lower weight HA molecule of 1100 kDA was found. Immunohistochemistry showed a larger amount of Wharton's jelly HA in DS UCs than in euploid UC. Real-time PCR analysis showed that HAS 2 and HYAL 2 were expressed at significant levels in all specimens. A higher expression of HAS 2 and a lower expression of HYAL 2 was found in the Wharton's jelly of DS fetuses compared to that of euploid fetuses at 14 weeks of gestation. On the contrary, at term HYAL 2 expression was higher in DS specimens than in those from euploid fetuses. Zymographic studies showed a similar behavior with a lower HYAL activity at early gestation and a higher HYAL activity at term gestation in DS UCs compared to euploid specimens. Therefore we can conclude that HA is more represented in DS UCs than in euploid UCs. A complex alteration of the HA metabolism characterized by an increased synthesis of lower weight HA molecules is a peculiarity of DS UCs. PMID- 15890267 TI - Decision making process in oncology practice: is the information available and what should it consist of? AB - In terms of systemic chemotherapy, the oncologist is often faced with the difficulty to discriminate between several options, at least according to evidence-based medicine. The aim of the present report was to assess to what extent the efficacy-related parameters required for the decision making process are reported in clinical trials. The analysis was restricted to lung, breast, colorectal and ovarian cancers. It included 135 phase II trials published in 1999, and 79 phase III trials published between 1991 and 2000. Response duration was mentioned in one-half and one-fourth of phases II and III trials, respectively. Only one-half of the trials reported time to progression (TTP). Finally, 28% of phase II and 44% of phase III trials reported RR, TTP and progression rates. The study indicates that the information available from reported clinical trials needs to be upgraded and homogenised in order to improve the decision making process in oncology. PMID- 15890268 TI - Predicting drug response and toxicity based on gene polymorphisms. AB - The sequencing of the human genome has allowed the identification of thousands of gene polymorphisms, most often single nucleotide polymorphims (SNP), which may play an important role in the expression level and activity of the corresponding proteins. When these polymorphisms occur at the level of drug metabolising enzymes or transporters, the disposition of the drug may be altered and, consequently, its efficacy may be compromised or its toxicity enhanced. Polymorphisms can also occur at the level of proteins directly involved in drug action, either when the protein is the target of the drug or when the protein is involved in the repair of drug-induced lesions. There again, these polymorphisms may lead to alterations in drug efficacy and/or toxicity. The identification of functional polymorphisms in patients undergoing chemotherapy may help the clinician prescribe the optimal drug combination or schedule and predict with more accuracy the response to these prescriptions. We have recorded in this review the polymorphisms that have been identified up till now in genes involved in anticancer drug activity. Some of them appear especially important in predicting drug toxicity and should be determined in routine before drug administration; this is the case of the most common variations of thiopurine methyltransferase for 6-mercaptopurine and of dihydropyrimidine dehydrogenase for fluorouracil. Other appear determinant for drug response, such as the common SNPs found in glutathione S-transferase P1 or xereoderma pigmentosum group D enzyme for the activity of oxaliplatin. However, confusion factors may exist between the role of gene polymorphisms in cancer risk or overall prognosis and their role in drug response. PMID- 15890269 TI - Chemotherapy in the primary treatment of cervical carcinoma. AB - Two major treatment modalities in cervical cancer include radiotherapy and surgery. In an attempt to improve the outcome, these modalities have been increasingly supplemented by chemotherapy. Chemotherapy can be combined with local therapies in various sequences. Of the two possible strategies using chemotherapy and radiotherapy (sequential or concomitant), the latter seems to be more effective. Platinum-based regimens applied concurrently with both definitive and post-operative radiation therapy were demonstrated to provide survival benefit in five of the six recently published randomised trials. The positive impact of chemotherapy added to radiotherapy has also been shown in a meta analysis including 1894 patients in 19 randomised studies. This strategy, however, is accompanied by increased early toxicity. The benefit of chemotherapy applied prior to surgery remains debatable. The role of new cytotoxic and biological substances, as well as agents combating tumour hypoxia, warrants further clinical investigation. PMID- 15890270 TI - Gastric cancer. AB - Gastric cancer is one of the most common cancers and one of the most frequent causes of cancer-related deaths. The incidence, diagnostic studies, and therapeutic options have undergone important changes in the last decades, but the prognosis for gastric cancer patients remains poor, especially in more advanced stages. Surgery is the mainstay of treatment of this disease, even if it is associated with a high rate of locoregional and distant recurrence. There is ongoing debate regarding the role of adjuvant treatment In advanced disease, palliation of symptoms, rather than cure, is the primary goal of patient management. Several combination therapies have been developed and have been examined in phase III trials; however, in most cases, they have failed to demonstrate a survival advantage over the reference arm. This review summarizes the most important recommendations for the management of patients with gastric cancer. PMID- 15890271 TI - Cancer of unknown primary (CUP). AB - Carcinoma of unknown primary (CUP) is one of the 10 most frequent cancers worldwide. It constitutes 3-5% of all human malignancies. Patients with CUP present with metastases without an established primary site. CUP manifests as an heterogenous group of mainly epithelial cancers recognised by distinct clinicopathological entities. The diagnostic work-up includes extensive histopathology investigations and modern imaging technology. Nevertheless, the primary tumour remains undetected most of the time. Certain clinicopathological CUP entities are considered as favourable subsets responding to systemic platinum based chemotherapy or managed by locoregional treatment. These subsets are: the poorly differentiated carcinomas involving the mediastinal-retroperitoneal nodes, peritoneal papillary serous adenocarcinomatosis in females, poorly differentiated neuroendocrine carcinomas, isolated axillary node adenocarcinomas in females or cervical nodal involvement by a squamous cell carcinoma. Patients who belong to the non-favourable subsets have a worse prognosis. PMID- 15890272 TI - Predicting oligomeric assemblies: N-mers a primer. AB - Multi-protein complexes play key roles in many biological processes. However, since the structures of these assemblies are hard to resolve experimentally, the detailed mechanism of how they work cooperatively in the cell has remained elusive. Similarly, recent advances on in silico prediction of protein-protein interactions have so far avoided this difficult problem. In this paper, we present a general algorithm to predict molecular assemblies of homo-oligomers. Given the number of N-mers and the 3D structure of one monomer, the method samples all the possible symmetries that N-mers can be assembled. Based on a scoring function that clusters the low free energy structures at each binding interface, the algorithm predicts the complex structure as well as the symmetry of the protein assembly. The method is quite general and does not involve any free parameters. The algorithm has been implemented as a public server and integrated to the protein-protein complex prediction server ClusPro. Using this application, we validated predictions for trimers, tetramers (discriminating between dimer of dimers and 4-fold symmetry structures), pentamers and hexamers (discriminating between trimer of dimers, dimer of trimers, and 6-fold symmetry structures), for a total of 107 assemblies. For 85% of the multimers, the server predicts the complex structure within an average rms deviation of 2A from the full crystal. For complexes that involve more than one binding interface, the cluster size at each surface provides a strong indication as to which interface forms first. With improving scoring functions and computer power, our multimer docking approach could be used as a framework to address the more general problem of multi-protein assemblies. PMID- 15890273 TI - Elongating modified conserved peptides eliminates their immunogenicity and protective efficacy against P. falciparum malaria. AB - Plasmodium falciparum malaria protein peptides were synthesised in the search for more effective routes for inducing a protective immune response against this deadly parasite and this information has been associated with such molecules' three-dimensional structure. These peptides had high red blood cell binding activity and their carboxy- and amino-terminal extremes were elongated for determining their immunogenic and protection-inducing activity against this disease in the Aotus monkey experimental model. 1H-NMR was used for analysing their three-dimensional structure; FAST ELISA, immunofluorescence antibody test, and Western blot were used for identifying their antibody inducing capacity and these previously immunised Aotus were inoculated with a highly infective P. falciparum strain to determine whether these elongated peptides were able to induce protection. This was aimed at establishing an association or correlation between long peptides' three-dimensional structure and their immunogenic and protection-inducing response in these monkeys. Peptides 20026 (25 residue), 20028 (30 residue), and 20030 (35 residues) were synthesised based on elongating the amino-terminal region of the 10022 highly immunogenic and protection-inducing modified peptide. 1H-NMR studies revealed that the first three had Classical type III beta-turn structures, different from the 20-amino acid long modified peptide 10022 which had a distorted type III beta-turn. Humoral immune response analysis showed that even when some antibodies could be generated against the parasite, none of the immunised Aotus could be protected with elongated peptides suggesting that elongating them eliminated modified peptide 10022 immunogenic and protection inducing capacity. PMID- 15890274 TI - Electron tomography reveals diverse conformations of integrin alphaIIbbeta3 in the active state. AB - We used electron tomography to determine the three-dimensional (3D) structure of integrin alphaIIbbeta3 in the active state. We found that we obtained better density maps when we reconstructed a 3D volume for each individual particle in the tilt series rather than to extract the particle-containing subvolumes from a 3D reconstruction of the entire specimen area. The 3D tomographic reconstructions of 100 particles revealed that activated alphaIIbbeta3 adopts many different conformations. An average of all the individual 3D reconstructions nicely accommodated the crystal structure of the alphaVbeta3 headpiece, confirming the locations assigned to the alpha- and beta-subunit in the density map. The most striking finding of our study is the structural flexibility of the lower leg of the beta-subunit as opposed to the conformational stability of the leg of the alpha-subunit. The good fit of the atomic structure of the betaI domain and the hybrid domain in the active state showed that the hybrid domain swings out, and most particles used for tomography are in the active state. Multivariate statistical analysis and classification applied to the set of 3D reconstructions revealed that more than 90% reconstructions are grouped into the classes that show the active state. Our results demonstrate that electron tomography can be used to classify complexes with a flexible structure such as integrins. PMID- 15890275 TI - Investigation of the morphology of intermediate filaments adsorbed to different solid supports. AB - Morphologically, glutaraldehyde-fixed and -dried intermediate filaments (IFs) appear flexible, and with a width of 8-12 nm when observed by electron microscopy. Sometimes, the filaments are even unraveled on the carbon-coated grid and reveal a protofilamentous architecture. In this study, we have used atomic force microscopy to further investigate the morphology of IFs in a more physiological environment. First, we have imaged hydrated glutaraldehyde-fixed IFs adsorbed to a graphite support. In such conditions, human vimentin and desmin IFs appeared compact with a height of 5-8 nm and revealed either a beading repeat or a helical morphology. Second, we have analyzed the architecture of hydrated vimentin, desmin, and neurofilament IFs adsorbed to mica, graphite, and hydrophilic glass without the presence of fixative. On mica, vimentin IFs had a height of only 3-5 nm, whereas desmin IFs appeared as 8-10 nm height filaments with a helical twist. Neurofilaments were 10-12 nm in height with a pronounced 30 50 nm beading along their length. On graphite, the different IFs were either not adsorbing properly or their architecture was modified yielding, for example, broad, flattened filaments. Finally, hydrophilic glass was the surface which seemed to best preserve the architecture of the three IFs, even if, in some cases, unraveled vimentin filaments were observed on this support. These results are straightening the idea that mature IFs are dynamic polymers in vitro and that IFs can be distinguished from each others by their physicochemical properties. PMID- 15890276 TI - Structural changes of DNA induced by mono- and binuclear cancer drugs. AB - The structural features of the drug-DNA adducts resulted from treatment of DNA with the platinum based mononuclear drug cisplatin and the binuclear drug [{trans PtCl(NH3)2}2H2N(CH2)4NH2]Cl2 or bis(platin) have been investigated by atomic force microscopy (AFM). Reduction in the contour length of the DNA fragments has been observed after cisplatin treatment while, compaction and aggregation are found to be the primary structural modifications following treatment with the binuclear drug. The intermolecular interaction upon bis(platin) treatment leads to observation of highly condense aggregates without a distinct sight of single isolated DNA molecule. These differences in drug binding indicate that unlike the mononuclear drug cisplatin, bis(platin) causes extensive interhelical/intermolecular cross-linking through its multiple linking sites. To our knowledge, this is the first report of a comparative AFM study to monitor the effects of a mono- and a binuclear platinum anti-cancer drug on DNA structure. These observations should provide clues towards explaining the distinct biological activities of the two drugs. PMID- 15890277 TI - A cylinder-shaped double ribbon structure formed by an amyloid hairpin peptide derived from the beta-sheet of murine PrP: an X-ray and molecular dynamics simulation study. AB - A structural model of the murine PrP small beta-sheet was obtained by synthesizing the RGYMLGSADPNGNQVYYRG peptide comprising the two beta-strands 127 133 and 159-164 linked by a four-residue sequence of high turn propensity. The DPNG turn sequence is a "short circuit" replacing the original protein sequence between the two strands. This 19-residue peptide spontaneously forms very long single fibrils as observed by electron microscopy. The X-ray diffraction patterns of a partially oriented sample reveals an average arrangement of the hairpin peptides into a structure which can be geometrically approximated by an empty core cylinder. The hairpins are oriented perpendicular to the cylinder axis and a 130 A helix period is observed. Based on X-ray diffraction constraints and on more indirect general protein structure considerations, a precise and consistent fibril model was built. The structure consists of two beta-sheet ribbons wound around a cylinder and assembled into a single fibril with a hairpin orientation perpendicular to the fibril axis. Subsequent implicit and explicit solvent molecular dynamics simulations provided the final structure at atomic resolution and further insights into the stabilizing interactions. Particularly important are the zipper-like network of polar interactions between the edges of the two ribbons, including the partially buried water molecules. The hydrophobic core is not optimally compact explaining the low density of this region seen by X-ray diffraction. The present findings provide also a simple model for further investigating the sequence-stability relationship using a mutational approach with a quasi-independent consideration of the polar and apolar interactions. PMID- 15890278 TI - Structure and function of the potent cyclic and linear melanocortin analogues. AB - The MC3R and MC4R proteins comprise two melanocortin receptor subtypes that are involved in obesity, with each protein displaying a unique mechanism of action. To enable the design of a selective drug candidate, the solution structures of four peptidyl analogues of the melanocyte stimulating hormones, NDP-MSH, NDP MSH(4-10) and two cyclic forms ([C5,C10]NDP-MSH(5-10), [C5,C10]NDP-MSH(5-11)), were characterized by two-dimensional nuclear magnetic resonance (NMR) spectroscopy and simulated annealing calculations. Using data from c-AMP assays in combination with structural analysis of melanocortin receptor/ligand models, we conclude that a lysine residue at the C-terminus of the His-Phe-Arg-Trp core sequence of melanocortin hormone is an important determinant for receptor selectivity in the both cyclic and linear MSH analogues. Our results suggest that side-chain orientation and charge-charge interactions with the ligand molecule play critical roles in receptor selectivity, whereas the overall backbone conformation or turn type contributes mainly to receptor binding. PMID- 15890279 TI - Crystal structure of nucleoside diphosphate kinase required for coleoptile elongation in rice (Oryza sativa L.). AB - Nucleoside diphosphate kinase (NDK) is a ubiquitous enzyme found in all organisms and cell types, and catalyzes the transfer of the phosphoryl group from a nucleoside triphosphate to a nucleoside diphosphate. The enzyme is involved in and required for coleoptile elongation in rice as the level of the rice NDK (rNDK) changes during seed germination and the early stages of seedling growth. The expression of rice NDK gene is up-regulated in the growing coleoptiles when the anaerobic stress persists. The rNDK structure determined at 2.5 A resolution consists of a four-stranded anti-parallel beta-sheet, of which the surfaces are partially covered with six alpha-helices; its overall and active site structures are similar to those of homologous enzymes except the major conformation variations of residue 132-138 regions, involving significant structural contacts. The model contains 148 residues of 149 residues in total and averaged 19 water molecules per monomer for 12 molecules in an asymmetric unit. A mold of 12 superimposed molecules shows that the alphaA-alpha2 area has greater variations and higher temperature factors, indicating the flexibility for a substrate entrance. Hexameric molecular packing in both crystal and solution implies that rNDK functions as hexamers. This rNDK structure, which is the first NDK structure from a higher plant system, provides the structural information essential to understand the functional significance of this enzyme during growth and development in both rice and other plants. PMID- 15890280 TI - The two-step mode of growth in the scleractinian coral skeletons from the micrometre to the overall scale. AB - It has been known since the 19th century that coral skeletons are built of aragonite crystals with taxonomy-linked arrangements, but the way by which each coral species controls this crystallization process remains an unsolved question. The problem became still more intriguing when it was shown that isotopic compositions of coral aragonite were subject to taxonomy-linked influences (the "vital effect"). On the other hand, presence of an organic component in coral skeletons is also long known, but localization of these compounds is admittedly restricted to particular structures called "centres of calcifications." Fibres, the largely predominant part of the coral skeletons, are usually considered as purely mineral units. In this paper, it is shown that in both "centres of calcification" and fibres, organic compounds are associated with the mineral material at a deep structural level. A series of variously scaled observations and localized measurements allow recognition of the presence of an organic component at the nanometre scale. Far from being a freely operating process, crystallization of coral fibres is thus permanently controlled by the polyp basal ectoderm through a cyclic two-step process acting at the micrometre-scale. The biomineralization cycle begins by secretion of a proteoglycan matrix. As the composition of these sugars-proteins assemblages has been shown taxonomy dependent, the hypothesis can be made that multiple and long recognized specificities of coral skeletons are linked to this biochemically driven crystallization process. Additionally, this new concept of the biomineralization process in coral skeletons provides us with an access to the long term evolution of the Scleractinia. Remarkably, results of a skeleton-based approach using microstructural criteria (i.e., the spatial relationships of "centres of calcification" and the three-dimensional arrangements of fibres), are consistent with a molecular phylogenetic analysis carried out on the same species. Clearly, at the overall ontogenic level, the two-step growth mode of coral skeletons is also a valuable tool to reconstruct the evolutionary history of Scleractinia. PMID- 15890281 TI - Classification and three-dimensional reconstruction of unevenly distributed or symmetry mismatched features of icosahedral particles. AB - Methods for the three-dimensional reconstruction of icosahedral particles, such as spherical viruses, from electron micrographs are well established. These methods take advantage of the 60-fold symmetry of the icosahedral group. Several features within these particles, however, may deviate from icosahedral symmetry. Examples include viral genomes, symmetry mismatched vertex proteins, unique DNA packaging vertices, flexible proteins, and proteins that are present at less than 100% occupancy. Such asymmetrically distributed features are smeared in the final density map when icosahedral symmetry is applied. Here, we describe a novel approach to classifying, analysing, and obtaining three-dimensional reconstructions of such features. The approach uses the orientation information derived from the icosahedral orientation search to facilitate multivariate statistical analysis and to limit the orientational degrees of freedom for reconstruction. We demonstrate the application of this approach to images of Kelp fly Virus. In this case, each virion may have two different types of fivefold vertex. We use our approach to produce independent reconstructions of the two types of vertex. PMID- 15890283 TI - Ten myths about decision-making capacity. AB - As a matter of practical reality, what role patients will play in decisions about their health care is determined by whether their clinicians judge them to have decision-making capacity. Because so much hinges on assessments of capacity, clinicians who work with patients have an ethical obligation to understand this concept. This article, based on a report prepared by the National Ethics Committee (NEC) of the Veterans Health Administration (VHA), seeks to provide clinicians with practical information about decision-making capacity and how it is assessed. A study of clinicians and ethics committee chairs carried out under the auspices of the NEC identified the following 10 common myths clinicians hold about decision-making capacity: (1) decision-making capacity and competency are the same; (2) lack of decision-making capacity can be presumed when patients go against medical advice; (3) there is no need to assess decision-making capacity unless patients go against medical advice; (4) decision-making capacity is an "all or nothing" phenomenon; (5) cognitive impairment equals lack of decision making capacity; (6) lack of decision-making capacity is a permanent condition; (7) patients who have not been given relevant and consistent information about their treatment lack decision-making capacity; (8) all patients with certain psychiatric disorders lack decision-making capacity; (9) patients who are involuntarily committed lack decision-making capacity; and (10) only mental health experts can assess decision-making capacity. By describing and debunking these common misconceptions, this article attempts to prevent potential errors in the clinical assessment of decision-making capacity, thereby supporting patients' right to make choices about their own health care. PMID- 15890286 TI - Barriers to managing pain in the nursing home: findings from a statewide survey. AB - OBJECTIVES: The objective of this study was to identify pain management demographics, perceived resources, and perceived barriers to adequately manage pain in the nursing home setting. DESIGN: Mailed survey. SETTING: All licensed Connecticut nursing homes. PARTICIPANTS: Directors of Nursing (DONs). MEASUREMENTS: Survey eliciting pain management demographics, perceived resources, and perceived barriers to adequately manage pain in respondents' nursing home. RESULTS: A total of 113 of 260 DONs (43%) responded to the survey. Respondents believed pain was suboptimally managed, particularly for residents with malignant and nonmalignant chronic pain. Perceived barriers to providing adequate pain management included lack of knowledge about pain management among nurses and physicians, lack of a standardized approach to treating pain, physicians' personal attitudes toward treating pain (eg, fear of addiction or overdose), lack of diagnostic precision in treating pain, and difficulty in choosing the right analgesic. Other barriers are also discussed, including low hospice enrollment of nursing home residents. CONCLUSION: Improving pain management in nursing homes requires improving provider knowledge and attitudes, enhancing diagnostic precision, standardizing pain treatment, and achieving an institutional commitment. Although responding DONs seemed aware of the need for improved pain management outcomes at their facilities, the required institutional commitment to accomplish this was not evidenced by these findings. PMID- 15890289 TI - End-of-life care in U.S. nursing homes: a review of the evidence. AB - OBJECTIVES: The purpose of this study was to systematically review the empiric evidence on end-of-life care in nursing homes in the United States The guiding research question for this review was what is the state of research evidence in end-of-life care in long-term care? DESIGN: We conducted a systematic review of the literature. DATA: The review was limited to published and indexed research in peer-reviewed journals in five major databases between 1995 and October 2002. RESULTS: The initial search yielded a total of 395 articles. The search was narrowed, focusing on nursing homes in the United States and empiric research. The result was 43 articles related to research in end-of-life care in American nursing homes. It was categorized into eight foci: prognosis, pain, hospice, hospitalization, advanced care planning, communication, family perceptions, and miscellaneous. CONCLUSION: There is a dearth of research published in end-of-life care in the nursing home setting. What is available is primarily descriptive. The empiric research only documents poor end-of-life care in U.S. nursing homes. Empiric evidence has grown in this area, but there is now a need for research of creative and innovative solutions aimed at improving the quality of end-of-life care in this setting. PMID- 15890292 TI - Management of constipation in residents with dementia: sorbitol effectiveness and cost. AB - OBJECTIVE: The objective of this report is to describe a cost-effective strategy for management of constipation in nursing home residents with dementia. DESIGN: We conducted a prospective observational quality improvement study of 41 residents with chronic constipation and receiving an osmotic laxative. Sorbitol was substituted for lactulose. SETTING: The study was conducted at a dementia special care unit at a Veterans Administration hospital. MEASUREMENT: We measured the number and amount of laxative use over a period of 4 weeks that were required to maintain regular bowel function. RESULTS: There was no difference in efficacy of lactulose and sorbitol. Use of additional laxatives was infrequent: Milk of Magnesia on approximately 10% of days/patient, bisacodyl suppository on 2% to 4% of days/patient, and Fleet enema only on 3 occasions. The cost of constipation management using routine administration of sorbitol and as-needed use of other laxatives was 27% to 55% lower than the cost of other constipation management strategies reported in the literature. CONCLUSION: Substitution of sorbitol for lactulose does not change efficacy of the treatment and decreases cost. Regular use of an osmotic laxative avoids the costs and discomforts of rectal laxatives. PMID- 15890294 TI - Quality indicators for the management of medical conditions in nursing home residents. AB - PURPOSE: The purpose of this study was to develop a set of specific care processes associated with better outcomes for general medical conditions identified as quality improvement targets for institutionalized vulnerable elders. METHODS: A national panel of nursing home experts used a modified-Delphi process to rate the validity (process linked to improved outcomes) and feasibility (of implementation and measurement) of candidate measures for depression, diabetes, hearing impairment, heart failure, hypertension, ischemic heart disease, osteoarthritis, osteoporosis, pneumonia, stroke, and vision impairment. Each quality indicator was written as an "if" statement, describing persons to whom the quality indicator applied followed by a "then" statement identifying the care process to be provided. A separate clinical committee reviewed the resulting set of indicators. RESULTS: One hundred fourteen quality indicators were identified across the 11 medical conditions. The quality indicators capture a broad range of medical care addressing assessment, management, and follow up. Fifty-five indicators (48%) were identical to quality measures for community-dwelling vulnerable elders. A limited number were rated as questionably feasible to implement or measure (6 and 2, respectively). Thirty eight (33%) would not be applied to measures of care quality for persons with advanced dementia or poor prognosis. CONCLUSIONS: Explicit care processes linked to improved nursing home outcomes for general medical conditions can be identified. Most of these care processes can be measured by medical records or interview. Nursing home quality measures for medical conditions must account for exclusions related to poor prognosis and advanced dementia. PMID- 15890297 TI - The effect of megestrol acetate on oral food and fluid intake in nursing home residents: a pilot study. AB - OBJECTIVES: The objective of this study was to evaluate the effect of megestrol acetate (Megace OS; Bristol-Myers Squibb, Princeton, NJ) on the oral food and fluid intake of nursing home (NH) residents under two conditions: usual NH care and optimal mealtime feeding assistance. DESIGN AND SETTING: We conducted a prospective, preliminary trial in four NHs. PARTICIPANTS: Participants (n = 17) were recruited from a larger study designed to assess nutritional care quality. Eligibility for the Megace OS trial required participants to consistently eat less than 75% of most meals under both usual NH care and optimal feeding assistance conditions at baseline. INTERVENTION: Megace OS, an oral liquid suspension of megestrol acetate, was given daily in a 400-mg dose for 63 days. MEASUREMENTS: Each participant's oral food and fluid intake was monitored weekly for 1 day (three meals) during which research staff conducted direct observations of usual NH care (weeks 1, 3, and 5 and day 63) or provided optimal feeding assistance (weeks 2, 4, and 6). Average total percent intake was compared from baseline across the assessment weeks of the trial under the two mealtime care conditions. RESULTS: Megace OS had a significant effect on oral food and fluid intake only under the optimal mealtime feeding assistance condition, in which average total percent eaten increased from 50% (+/-15%) at baseline to 63% (+/ 14%) post-63 days of the trial. There was no change in participants' oral food and fluid intake under the usual NH care condition (average total percent intake at baseline 43% +/- 12% vs. 43% +/- 20% post-63 days). CONCLUSION: The results of this preliminary study suggest that Megace OS is not an effective nutritional intervention to increase oral intake under usual NH care conditions, which is often characterized by inadequate feeding assistance. However, Megace OS in combination with optimal mealtime feeding assistance does significantly increase oral intake in a frail NH sample at high risk for weight loss. PMID- 15890298 TI - Feasibility of quality indicators for the management of geriatric syndromes in nursing home residents. AB - PURPOSE: The assessment and management of dementia, falls and mobility disorders, malnutrition, end-of-life issues, pressure ulcers, and urinary incontinence have been identified as important quality improvement targets for vulnerable elders residing in nursing homes. This study aimed to identify valid and feasible measures of specific care processes associated with improved outcomes for these conditions. METHODS: Nine experts in nursing home (NH) care participated in a modified Delphi process to evaluate potential quality indicators (QIs) for care in NHs. Panelists met and discussed potential indicators before completing confidential ballots rating validity (process associated with improved outcomes), feasibility of measurement (with charts or interviews), feasibility of implementation (given staffing resources in average community NHs), and importance (expected benefit and prevalence in NHs). The NH panel's median votes were used to identify a final set of QIs that were subsequently reviewed by a clinical oversight committee. RESULTS: Sixty-eight geriatric syndrome QIs were identified as valid and important in NH populations. Panelists assessed 12 (18%) of these QIs as having questionable feasibility to implement in average community nursing homes trying to provide quality care. Nine (13%) would not be included in systems assessing quality of care for persons with advanced dementia or poor prognosis. CONCLUSIONS: Steps of care critical to the assessment and management of geriatric syndromes in NHs were identified. Feasibility is an important issue for a significant number of these, indicating that much remains to be done to design systems that efficiently and reliably implement these care processes. PMID- 15890300 TI - Barriers to providing osteoporosis care in skilled nursing facilities: perceptions of medical directors and directors of nursing. AB - OBJECTIVES: The objectives of this study were to identify the barriers to osteoporosis clinical practice guideline use perceived by Medical Directors (MED DIR) and Directors of Nursing (DON) in skilled nursing facilities; and to describe differences in the perceptions of MED DIRs and DONs. DESIGN: The authors conducted a cross-sectional national survey. PARTICIPANTS: This study consisted of a random national sample of MED DIRs (n = 1300) and DONs (n = 1300) belonging to the American Medical Directors Association or the National Association of Directors of Nursing Administration in Long-term Care. MEASUREMENTS: A 24-item survey using a five-point Likert scale was developed. The survey measured agreement to questions in four domains (provider factors, guideline characteristics, patient factors, environmental factors) and 10 content areas (problem acknowledgment, patient/family concern, patient/family compliance, testing availability, safety, reimbursement, regulatory oversight, staff knowledge/time/ability, belief in guidelines, and malpractice liability). Response distributions to each item were plotted and differences between MED DIRs and DONs were tested. RESULTS: Survey response rates were 40% for MED DIRs and 48% for DONs. Respondents strongly agreed that fractures are a problem in their facilities and that osteoporosis guidelines are useful and cost-beneficial (mean responses > or = 4.0). A large proportion of respondents (at least 40% of the sample) identified multiple patient comorbidities, reimbursement issues, length of stay, and regulatory oversight as barriers to providing osteoporosis care. Respondents did not believe that patient and family acceptance, testing availability, staff time, staff self-efficacy, or concerns about bisphosphonate safety were barriers to osteoporosis care. DONs were more likely than MED DIRs to believe that patients and families are concerned about fractures, whereas MED DIRs were more likely to endorse length of stay, staffing issues, and regulatory oversight as influencing treatment decisions. Years of practice and facility size, but not formal geriatrics training, significantly influenced responses. CONCLUSION: Perceived barriers to implementing osteoporosis guidelines differ between facilities and between MED DIRs and DONs. Identification of these barriers could facilitate quality improvement initiatives and improve the quality of osteoporosis care. PMID- 15890302 TI - Corticosteroids and chronic obstructive pulmonary disease in the nursing home. PMID- 15890304 TI - Can we prevent aspiration pneumonia in the nursing home? AB - Aspiration pneumonia is a significant cause of morbidity, hospitalization, and mortality in the nursing home population. Patients who aspirate have three times higher mortality than patients who do not aspirate. We discuss the factors known to increase the risk of aspiration and its consequences, and recognize some of the preventive measures for aspiration pneumonia. We suggest approaches to decrease the risk of this very prevalent syndrome. PMID- 15890306 TI - Falls in the nursing home: Are they preventable? AB - INTRODUCTION: Falls are prevalent in elderly patients residing in nursing homes, with approximately 1.5 falls occurring per nursing home bed-years. Although most are benign and injury-free, 10% to 25% result in hospital admission and/or fractures. Primary care providers for nursing home residents must therefore aim to reduce both the fall rate as well as the rate of fallrelated morbidity in the long-term care setting. Interventions have been demonstrated to be successful in reducing falls in community-dwelling elderly patients. However, less evidence supports the efficacy of fall prevention in nursing home residents. METHODS: The authors conducted a Medline search using the key words Falls and Nursing Homes. RESULTS: Several studies examined the efficacy of multifaceted intervention programs on reducing falls in nursing homes with varied results. Components of these intervention programs include: environmental assessment, assistive device evaluation and modification, medication changes, gait assessment and training, staff education, exercise programs, hip protector use, and blood pressure evaluation. Current literature supports the use of environmental assessment and intervention in reducing falls in nursing homes, and demonstrates an association between certain medications and falls. However, there are no studies that examine the effect of medication adjustments on fall rates. Also, the literature does not strongly suggest that exercise programs are effective in fall reduction. Although not effective in reducing fall rates, the use of hip protectors appears to result in less fall-related morbidity. CONCLUSION: More studies must be done to clarify the effects of high-risk medication reduction, the optimal nature and intensity of exercise programs, and patient targeting criteria to maximize the effectiveness of nursing home fall prevention programs. Based on the current literature, an effective multifaceted fall prevention program for nursing home residents should include risk factor assessment and modification, staff education, gait assessment and intervention, assistive device assessment and optimization, as well as environmental assessment and modification. Although there is no association between the use of hip protectors and fall rates, their use should be encouraged because the ultimate goal of any fall prevention program is to prevent fall-related morbidity. PMID- 15890308 TI - Quality palliative care in long-term care settings. AB - It is paramount that physicians and midlevel practitioners who care for residents in long-term care facilities be able to provide high-quality comfort care to their patients, the majority of whom are frail and suffering from chronic and progressive diseases. Physicians must be knowledgeable in the assessment, prevention, and relief of patients' physical, emotional, and spiritual distress, as well as develop appropriate attitudes, knowledge, and skills to care for patients who are in the last years of life. The provision of high-quality palliative care is the essence of long-term care medicine. PMID- 15890310 TI - [Gastric carcinoid tumours. Is there a place for antrectomy?]. AB - Gastric carcinoid tumours are classified in 3 types depending on whether they are sporadic (type III), or they are associated with a chronic atrophic gastritis (type I) or a multiple endocrine neoplasia type I-associated Zollinger-Ellison syndrome (type II). For type I tumours, the role of antrectomy, which aims to suppress the causative hypergastrinemia, has not been determined. AIM: To determine, from literature review the role of antrectomy in the management of type I gastric carcinoid tumours. METHODS: Bibliographic study searching for published observations of antrectomy for type I gastric carcinoid tumours. Data regarding postoperative evolution of gastrinemia and carcinoid tumours were collected. RESULTS: Thirty-eight published cases were identified. Preoperative gastrinemia was elevated in the 32 patients in whom it was measured. It came to normal ranges in the 19 patients in whom it was postoperatively assessed. With a mean follow-up of 34 months (1 to 120), disappearance of carcinoid tumours was observed in 27 of 38 patients (71%), the 11 others having tumour recurrence or persistence. When postoperatively assessed, hyperplasia of fundic enterochromaffine-like cells persisted in 7 patients, regressed in 4 and disappeared in the 6 others. No antrectomy-related complication was reported. CONCLUSION: Antrectomy can be considered as a worthwhile alternative for the treatment of gastric carcinoid tumours related to chronic atrophic gastritis and hypergastrinemia. PMID- 15890311 TI - The effect of a hyposmotic shock and purinergic agonists on K+(Rb+) efflux from cultured human breast cancer cells. AB - The effect of a hyposmotic shock and extracellular ATP on the efflux of K(+)(Rb(+)) from human breast cancer cell lines (MDA-MB-231 and MCF-7) has been examined. A hyposmotic shock increased the fractional efflux of K(+)(Rb(+)) from MDA-MB-231 cells via a pathway which was unaffected by Cl(-) replacement. Apamin, charybdotoxin or removing extracellular Ca(2+) had no effect on volume-activated K(+)(Rb(+)) efflux MDA-MB-231 cells. An osmotic shock also stimulated K(+)(Rb(+)) efflux from MCF-7 cells but to a much lesser extent than found with MDA-MB-231 cells. ATP-stimulated K(+)(Rb(+)) efflux from MDA-MB-231 cells in a dose dependent fashion but had little effect on K(+)(Rb(+)) release from MCF-7 cells. ATP-stimulated K(+)(Rb(+)) efflux was only inhibited slightly by replacing Cl(-) with NO(3)(-). Removal of external Ca(2+) during treatment with ATP reduced the fractional efflux of K(+)(Rb(+)) in a manner suggesting a role for cellular Ca(2+) stores. Charybdotoxin, but neither apamin nor iberiotoxin, inhibited ATP stimulated K(+)(Rb(+)) release from MDA-MB-231 cells. Suramin inhibited the ATP activated efflux of K(+)(Rb(+)). UTP also stimulated K(+)(Rb(+)) efflux from MDA MB-231 cells whereas ADP, AMP and adenosine were without effect. A combination of an osmotic shock and ATP increased the fractional efflux of K(+)(Rb(+)) to a level greater than the sum of the individual treatments. It appears that the hyposmotically-activated and ATP-stimulated K(+) efflux pathways are separate entities. However, there may be a degree of 'crosstalk' between the two pathways. PMID- 15890312 TI - Giant liposomes in physiological buffer using electroformation in a flow chamber. AB - We describe a method to obtain giant liposomes (diameter 10-100 microm) in solutions of high ionic strength to perform a membrane-binding assay under physiological conditions. Using electroformation on ITO electrodes, we formed surface-attached giant liposomes in solutions of glycerol in a flow chamber and then introduced solutions of high ionic strength (up to 2 M KCl) into this chamber. The ionic solution exchanged with the isoosmolar glycerol solution inside and outside the liposomes. An initial mismatch in index of refraction between the inside and outside of liposomes allowed for the observation of solution replacement. Ions and small polar molecules exchanged into and out of surface-attached liposomes within minutes. In contrast, liposomes formed in solutions of macromolecules retained molecules larger than 4 kDa, allowing for encapsulation of these molecules for hours or days even if the solution outside the liposomes was exchanged. We propose that solutes entered liposomes through lipid tubules that attach liposomes to the film of lipids on the surface of the ITO electrode. The method presented here makes it straightforward to perform flow through binding assays on giant liposomes under conditions of physiological ionic strength. We performed a membrane-binding assay for annexin V, a calcium dependent protein that binds to phosphatidylserine (PS). The binding of annexin V depended on the concentration of PS and decreased as ionic strength increased to physiological levels. PMID- 15890313 TI - Repeated checking really does cause memory distrust. AB - Memory phenomena associated with obsessive-compulsive disorder (OCD) have received increased attention in the recent literature. Some debate remains about whether OCD is characterized by deficits in memory per se, or by poor memory confidence. Following from a recent study that demonstrated memory distrust results from repeated checking of a virtual computerized stove, we asked 50 undergraduate students to repeatedly turn on, turn off and check either a real kitchen stove (relevant checking) or a real kitchen faucet (irrelevant checking) in a standardized, ritualized manner. All participants completed a final check of the stove following these 19 checking trials. Results indicated that following repeated relevant checking, participants reported significantly reduced memory confidence, vividness and detail. Repeated irrelevant checking did not produce these decreases. Results are discussed in terms of cognitive-behavioural formulations of OCD and in terms of the effects of repetition on memory and metamemory. PMID- 15890314 TI - Improving academic performance and mental health through a stress management intervention: outcomes and mediators of change. AB - Two hundred and nine pupils were randomly allocated to either a cognitive behaviourally based stress management intervention (SMI) group, or a non intervention control group. Mood and motivation measures were administered pre and post intervention. Standardized examinations were taken 8-10 weeks later. As hypothesized, results indicated that an increase in the functionality of pupils' cognitions served as the mechanism by which mental health improved in the SMI group. In contrast, the control group demonstrated no such improvements. Also, as predicted, an increase in motivation accounted for the SMI group's significantly better performance on the standardized, academic assessments that comprise the United Kingdom's General Certificate of Secondary Education. Indeed, the magnitude of this enhanced performance was, on average, one-letter grade. Discussion focuses on the theoretical and practical implications of these findings. PMID- 15890315 TI - A part of cholinergic fibers in mouse superior cervical ganglia contain GABA or glutamate. AB - The localizations and functions of glutamate and GABA, the major amino acid neurotransmitters in the central nervous system, are still unclear in the peripheral nervous system. We immunohistochemically double-stained mouse superior cervical ganglia with antibodies for the vesicular acetylcholine transporter (VAchT), GAD65, the vesicular glutamate transporters 1-3 (VGluTs1-3), the marker of the sympathetic preganglionic neuron (SPN), GABAergic, and glutamatergic terminals, respectively. All GAD65-positive terminals showed VAchT immunoreactivity, indicating that GABAergic fibers originate from SPNs. VGluT2 immunoreactive terminals showing colocalization with VAchT were observed, but VGluT1 and 3 immunoreactive terminals were not. Colocalization of GAD65 and VGluT2 was rarely found. All VGluT2-immunopositive terminals were also immunopositive for neuronal nitric oxide synthase (nNOS), a marker for the subpopulation of the SPNs, while about half of the GABA-immunopositive fibers were immunopositive for nNOS. The origin of these fibers was discussed. PMID- 15890316 TI - The NMDA receptor NR2B subunit contributes to epileptogenesis in human cortical dysplasia. AB - Cortical dysplasia (CD) is often associated with pharmacoresistant epilepsy. Previous studies showed increased expression of the NMDA receptor subunit NR2B in dysplastic and epileptic human neocortex. We tested the hypothesis that differential increase of NR2B constitutes an epileptogenic mechanism in humans. Dysplastic neocortex and lateral temporal lobe regions resected for treatment of pharmacoresistant seizures were processed for electrophysiological, histological, and immunocytochemical studies. Assignment to the "dysplastic" (n = 8) and "non dysplastic" (n = 8) groups was based on histology. Neurons in "dysplastic" samples differentially stained for NR2B. Western blot (n = 6) showed an immunoreactive band for NR2B in three out of four "dysplastic" samples. Epileptiform field potentials (EFP) were elicited in vitro by omission of magnesium from the bath. EFP in "dysplastic" slices were characterized by multiple afterdischarges, occurring at a significantly higher repetition rate than EFP in non-dysplastic slices. The NR2B-specific NMDA receptor inhibitor ifenprodil (10muM) suppressed EFP in dysplastic slices. In non-dysplastic slices, burst repetition rate did not change with ifenprodil application. In both dysplastic and non-dysplastic slices, EFP were suppressed by a non-specific NMDAR antagonist (APV) or AMPA receptor antagonist (CNQX). These results provide additional evidence that the differential expression of NR2B in dysplastic human neocortex may play a role in the expression of in-situ epileptogenesis in human CD. NR2B may constitute a target for new diagnostic and pharmacotherapeutic approaches. PMID- 15890317 TI - Endothelial nitric oxide synthase upregulation in the guinea pig organ of Corti after acute noise trauma. AB - Endothelial nitric oxide synthase (eNOS) upregulation was identified 60 h after acute noise trauma in morphologically intact cells of the reticular lamina in the organ of Corti of the guinea pig in the second turn of the cochlea. Using gold coupled anti-eNOS antibodies and electron microscopy, it was shown that eNOS expression was upregulated in all cell areas and cell types except inner hair cells. Furthermore, eNOS was found in the organelle-free cytoplasm and in mitochondria of various cell types. The density of eNOS in mitochondria was considerably higher compared with the surrounding cytoplasm. Since eNOS activity is regulated by calcium, the eNOS detection was combined with calcium precipitation, a method for visualizing intracellular Ca2+ distribution. After acute noise trauma, intracellular Ca2+ was increased in all cell types and cell areas except in outer hair cells. Comparing the distribution patterns of eNOS and calcium, significantly elevated levels (P < 0.0001) of eNOS were detected within a 100 nm radius near calcium precipitates in all cuticular structures as well as microtubule-rich regions and Deiters' cells near Hensen cells. The observed colocalization lends support to the postulated mechanism of eNOS activation by Ca2+. eNOS upregulation after acute noise trauma might therefore be part of an induced stress response. The eNOS upregulation in cell areas with numerous microtubule- and actin-rich structures is discussed with respect to possible cytoskeleton-dependent processes in eNOS regulation. PMID- 15890318 TI - Signaling pathways in the nitric oxide and iron-induced dopamine release in the striatum of freely moving rats: role of extracellular Ca2+ and L-type Ca2+ channels. AB - We showed previously that exogenous iron potentiated nitric oxide (NO) donor induced release of striatal dopamine (DA) in freely moving rats, using microdialysis. In this study, the increase in dialysate DA induced by intrastriatal infusion of the NO-donor 3-morpholinosydnonimine (SIN-1, 1.0 mM for 180 min) was scarcely affected by Ca2+ omission. N-methyl-d-glucamine dithiocarbamate (MGD) is a thiol compound whose NO trapping activity is potentiated by iron(II). Intrastriatal co-infusion of MGD either alone or associated with iron(II), however, potentiated SIN-1-induced increases in dialysate DA. In contrast, co-infusion of the NO trapper 4-(carboxyphenyl) 4,4,5,5-tetramethylimidazole-1-oxyl 3-oxide (carboxy-PTIO) significantly attenuated the increase in dialysate DA induced by SIN-1 (5.0 mM for 180 min). SIN-1+MGD+iron(II)-induced increases in dialysate DA were inhibited by Ca2+ omission or co-infusion of either deferoxamine or the L-type (Ca(v) 1.1-1.3) Ca2+ channel inhibitor nifedipine; in contrast, the increase was scarcely affected by co-infusion of the N-type (Ca(v) 2.2) Ca2+ channel inhibitor omega-conotoxin GVIA. These results demonstrate that exogenous NO-induced release of striatal DA is independent on extracellular Ca2+; however, in presence of the NO trapper MGD, NO may preferentially react with either endogenous or exogenous iron to form a complex which releases striatal DA with an extracellular Ca2+-dependent and nifedipine-sensitive mechanism. PMID- 15890319 TI - Central injection of hypertonic saline activates angiotensin II-sensitive neurons in the anterior hypothalamic area of rats. AB - We have previously reported that microinjection of angiotensin II into the anterior hypothalamic area (AHA) produces pressor responses and that angiotensin II-sensitive neurons in the AHA are tonically activated by endogenous angiotensins in rats. Central injection of hypertonic saline causes pressor responses via release of angiotensins in brain. In this study, we examined whether angiotensin II-sensitive neurons in the AHA are responsive to intracerebroventricular injection of hypertonic saline and whether endogenous angiotensins in the AHA are involved in the central hypertonic saline-induced pressor response. Male Wistar rats were anesthetized and artificially ventilated. Extracellular potentials were recorded from single neurons in the AHA. Intraventricular injection of hypertonic saline increased the neural activity of angiotensin II-sensitive neurons, whereas pressure application of hypertonic saline onto angiotensin II-sensitive neurons themselves did not affect their neural activities. The intraventricular hypertonic saline-induced increase of unit activity of AHA neurons was inhibited by pressure application of the angiotensin AT1 receptor antagonist losartan onto the same neurons. The hypertonic saline-induced increase of unit firing was also blocked by intraventricular injection of the amiloride-sensitive sodium channel blocker benzamil. In conscious rats, intraventricular injection of hypertonic saline produced pressor responses, and the hypertonic saline-induced pressor response was inhibited by bilateral microinjection of losartan into the AHA. Repeated intraventricular injection of hypertonic saline caused an increase in the release of angiotensins in the AHA of anesthetized rats. These findings indicate that intracerebroventricular injection of hypertonic saline increases neural activity of angiotensin II-sensitive neurons trans-synaptically via endogenous angiotensins in the AHA. In addition, these findings also indicate that the intracerebroventricular injection of hypertonic saline produces a pressor response at least partly via release of angiotensins in the AHA. PMID- 15890320 TI - Structure of a lactic acid ether-containing and glycerol phosphate-containing O polysaccharide from Proteus mirabilis O40. AB - An O-polysaccharide was isolated by mild acid hydrolysis from the lipopolysaccharide of Proteus mirabilis O40 and studied by NMR spectroscopy, including 2D 1H, 1H COSY, TOCSY, ROESY, and 1H, 13C HMQC experiments, along with chemical methods. The polysaccharide was found to contain an ether of GlcNAc with lactic acid and glycerol phosphate in the main chain and to have the following structure: --> 3)-beta-D-GlcpNAc4(R-Lac)-(1 --> 3)-alpha-D-Galp-(1 --> 3)-D-Gro-1 P-(O --> 3)-beta-D-GlcpNAc-(1 --> where D-GlcpNAc4(R-Lac) stands for 2-acetamido 4-O-[(R)-1-carboxyethyl]-2-deoxy-D-glucose. This structure is unique among the known structures of the Proteus O-polysaccharides, which is in agreement with the classification of the strain studied into a separate O-serogroup. A serological relatedness of P. mirabilis O40 with some other Proteus strains was revealed and discussed in view of the O-polysaccharide structures. PMID- 15890321 TI - Selective deprotection of terminal isopropylidene acetals and trityl ethers using HClO4 supported on silica gel. AB - Terminal isopropylidene acetals are selectively cleaved to the corresponding 1,2 diols in good to excellent yields in 6-24 h at room temperature by using the 'HClO4.SiO2' reagent system. Likewise, trityl ethers are readily cleaved to the corresponding alcohols in good to excellent yields within 2-3 h at room temperature. Work-up involves merely filtration of the reagent followed by purification of the crude product. PMID- 15890322 TI - Short QT syndrome. AB - The short QT syndrome constitutes a new clinical entity that is associated with a high incidence of sudden cardiac death, syncope, and/or atrial fibrillation even in young patients and newborns. Patients with this congenital electrical abnormality are characterized by rate-corrected QT intervals<320 ms. Missense mutations in KCNH2 (HERG) linked to a gain-of-function of the rapidly activating delayed-rectifier current I(Kr) have been identified in the first two reported families with familial sudden cardiac death. Recently, two further gain-of function mutations in the KCNQ1 gene encoding the alpha-subunit of the KvLQT1 (I(Ks)) channel and in the KCNJ2 gene encoding the strong inwardly rectifying channel protein Kir2.1 confirmed a genetically heterogeneous disease. The possible substrate for the development of ventricular tachyarrhythmias may be a significant transmural dispersion of the repolarisation due to a heterogeneous abbreviation of the action potential duration. The implantable cardioverter defibrillator is the therapy of choice in patients with syncope and a positive family history of sudden cardiac death. However, ICD therapy in patients with a short QT syndrome has an increased risk for inappropriate shock therapies due to possible T wave oversensing. The impact of sotalol, ibutilide, flecainide, and quinidine on QT prolongation has been evaluated, but only quinidine effectively suppressed gain-of-function in I(Kr) with prolongation of the QT interval. In patients with a mutation in HERG, it rendered ventricular tachycardias/ventricular fibrillation non-inducible and restored the QT interval/heart rate relationship towards a normal range. It may serve as an adjunct to ICD therapy or as a possible alternative treatment, especially for children and newborns. PMID- 15890323 TI - Brugada syndrome and fever: genetic and molecular characterization of patients carrying SCN5A mutations. AB - OBJECTIVE: Brugada syndrome (BrS) is characterized by ventricular tachyarrhythmias leading to sudden cardiac death and is caused, in part, by mutations in the SCN5A gene encoding the sodium channel Na(v)1.5. Fever can trigger or exacerbate the clinical manifestations of BrS. The aim of this work was to characterize the genetic and molecular determinants of fever-dependent BrS. METHODS: Four male patients with typical BrS ST-segment elevation in V1-V3 or ventricular arrhythmias during fever were screened for mutations in the SCN5A gene. Wild-type (WT) and mutant Na(v)1.5 channels were expressed in HEK293 cells. The sodium currents (I(Na)) were analysed using the whole-cell patch clamp technique at various temperatures. Protein expression of WT and mutant channels was studied by Western blot experiments. RESULTS: Two mutations in SCN5A, L325R and R535X, were identified. Expression of the two mutant Na(v)1.5 channels in HEK293 cells revealed in each case a severe loss-of-function. Upon the increase of temperature up to 42 degrees C, we observed a pronounced acceleration of Na(v)1.5 activation and fast inactivation kinetics. Cardiac action potential modelling experiments suggest that in patients with reduced I(Na), fever could prematurely shorten the action potential by virtue of its effect on WT channels. Further experiments revealed that L325R channels are likely misfolded, since their function could be partially rescued by mexiletine or curcumin. In co expression experiments, L325R channels interfered with the proper function of WT channels, suggesting that a dominant negative phenomenon may underlie BrS triggered by fever. CONCLUSIONS: The genetic background of BrS patients sensitive to fever is heterogeneous. Our experimental data suggest that the clinical manifestations of fever-exacerbated BrS may not be mutation specific. PMID- 15890324 TI - Enhanced expression of cyclooxygenase-2 and prostaglandin E2 in response to endotoxin after trauma is dependent on MAPK and NF-kappaB mechanisms. AB - Macrophage prostaglandin E2 (PGE2) production is important in cellular immune suppression and in affecting the potential development of sepsis after trauma. We hypothesized that macrophage PGE2 production after trauma is regulated by mitogen activated protein kinase (MAPK) and nuclear factor kappa B (NF-kappaB). Mice were subjected to trauma and splenic macrophages isolated 7 days later. Macrophages from traumatized mice showed increased cyclooxygenase-2 (COX-2) mRNA, protein expression, and PGE2 production compared with controls. Increased phosphorylation of extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), and p38 kinase was observed in macrophages from traumatized mice. Pharmacologic inhibition of MAPK blocked trauma-induced COX-2 expression, and PGE2 production. Trauma macrophages showed increased IkappaBalpha phosphorylation and NF-kappaB binding to DNA. Inhibiting IkappaBalpha blocked trauma-induced NF-kappaB activity, COX-2 expression and PGE2 production. This suggests that trauma-induced PGE2 production is mediated through MAPK and NF-kappaB activation and offers potential for modifying the macrophages' responses following injury. PMID- 15890325 TI - Nanotechnology in clinical laboratory diagnostics. AB - Nanotechnology-the creation and utilization of materials, devices, and systems through the control of matter on the nanometer-has been applied to molecular diagnostics. This article reviews nanobiotechnologies that are clinically relevant and have the potential to be incorporated in clinical laboratory diagnosis. Nanotechnologies enable the diagnosis at single cell and molecule level and some of these can be incorporated in the current molecular diagnostics such as biochips. Nanoparticles, such as gold nanoparticles and quantum dots, are the most widely used but various other nanotechnologies for manipulation at nanoscale as well as nanobiosensors are reviewed. These technologies will extend the limits of current molecular diagnostics and enable point-of-care diagnosis as well as the development of personalized medicine. Although the potential diagnostic applications are unlimited, most important current applications are foreseen in the areas of biomarker research, cancer diagnosis and detection of infectious microorganisms. PMID- 15890326 TI - Utilization of artificial neural networks and autoregressive modeling in diagnosing mitral valve stenosis. AB - This research is concentrated on the diagnosis of mitral heart valve stenosis through the analysis of Doppler Signals' AR power spectral density graphic with the help of ANN. Multilayer feedforward ANN trained with a Levenberg Marquart backpropagation algorithm was implemented in the MATLAB environment. Correct classification of 94% was achieved, whereas 4 false classifications have been observed for the test group of 68 subjects in total. The designed classification structure has about 97.3% sensitivity, 90.3% specifity and positive prediction is calculated to be 92.3%. The stated results show that the proposed method can make an effective interpretation. PMID- 15890327 TI - Discontinuous doppler signals simulating respiratory misregistration: effect on autoregressive frequency spectra. AB - In this study, we have produced discontinuous Doppler signals of carotid artery and internal jugular vein, simulating respiratory misregistration. The aim of the study is to observe the effect of signal discontinuity and its duration on power spectral density vs. frequency graphs obtained by Autoregressive Modeling. The signals were recorded from ten male volunteers. Signal interruption was performed by moving the sampling volume in and out of the vessel bidirectionally. To estimate the effect of on-line recording time and signal discontinuity on frequency spectra, we have worked on a control data of 30s with continuous signal, and three sets of data with artificially interrupted signals of 30, 60 and 90s duration. Maximum power spectral density, area under the power spectral density, and frequency level corresponding to maximum power spectral density were calculated on frequency spectra. The frequency level corresponding to maximum power spectral density provides the most statistically stable finding in our preliminary data. The signal duration of the signal had no significant effect on the statistical stability of the frequency level. PMID- 15890328 TI - 15-zinc finger protein Bloody Fingers is required for zebrafish morphogenetic movements during neurulation. AB - A novel zebrafish gene bloody fingers (blf) encoding a 478 amino acid protein containing fifteen C(2)H(2) type zinc fingers was identified by expression screening. As determined by in situ hybridization, blf RNA displays strong ubiquitous early zygotic expression, while during late gastrulation and early somitogenesis, blf expression becomes transiently restricted to the posterior dorsal and lateral mesoderm. During later somitogenesis, blf expression appears only in hematopoietic cells. It is completely eliminated in cloche, moonshine but not in vlad tepes (gata1) mutant embryos. Morpholino (MO) knockdown of the Blf protein results in the defects of morphogenetic movements. Blf-MO-injected embryos (morphants) display shortened and widened axial tissues due to defective convergent extension. Unlike other convergent extension mutants, blf morphants display a split neural tube, resulting in a phenotype similar to the human open neural tube defect spina bifida. In addition, dorsal ectodermal cells delaminate in blf morphants during late somitogenesis. We propose a model explaining the role of blf in convergent extension and neurulation. We conclude that blf plays an important role in regulating morphogenetic movements during gastrulation and neurulation while its role in hematopoiesis may be redundant. PMID- 15890329 TI - Gli function is essential for motor neuron induction in zebrafish. AB - The Gli family of zinc-finger transcription factors mediates Hedgehog (Hh) signaling in all vertebrates. However, their roles in ventral neural tube patterning, in particular motor neuron induction, appear to have diverged across species. For instance, cranial motor neurons are essentially lost in zebrafish detour (gli1(-)) mutants, whereas motor neuron development is unaffected in mouse single gli and some double gli knockouts. Interestingly, the expression of some Hh-regulated genes (ptc1, net1a, gli1) is mostly unaffected in the detour mutant hindbrain, suggesting that other Gli transcriptional activators may be involved. To better define the roles of the zebrafish gli genes in motor neuron induction and in Hh-regulated gene expression, we examined these processes in you-too (yot) mutants, which encode dominant repressor forms of Gli2 (Gli2(DR)), and following morpholino-mediated knockdown of gli1, gli2, and gli3 function. Motor neuron induction at all axial levels was reduced in yot (gli2(DR)) mutant embryos. In addition, Hh target gene expression at all axial levels except in rhombomere 4 was also reduced, suggesting an interference with the function of other Glis. Indeed, morpholino-mediated knockdown of Gli2(DR) protein in yot mutants led to a suppression of the defective motor neuron phenotype. However, gli2 knockdown in wild-type embryos generated no discernable motor neuron phenotype, while gli3 knockdown reduced motor neuron induction in the hindbrain and spinal cord. Significantly, gli2 or gli3 knockdown in detour (gli1(-)) mutants revealed roles for Gli2 and Gli3 activator functions in ptc1 expression and spinal motor neuron induction. Similarly, gli1 or gli3 knockdown in yot (gli2(DR)) mutants resulted in severe or complete loss of motor neurons, and of ptc1 and net1a expression, in the hindbrain and spinal cord. In addition, gli1 expression was greatly reduced in yot mutants following gli3, but not gli1, knockdown, suggesting that Gli3 activator function is specifically required for gli1 expression. These observations demonstrate that Gli activator function (encoded by gli1, gli2, and gli3) is essential for motor neuron induction and Hh-regulated gene expression in zebrafish. PMID- 15890330 TI - The role of GDNF in patterning the excretory system. AB - Mesenchymal-epithelial interactions are an important source of information for pattern formation during organogenesis. In the developing excretory system, one of the secreted mesenchymal factors thought to play a critical role in patterning the growth and branching of the epithelial ureteric bud is GDNF. We have tested the requirement for GDNF as a paracrine chemoattractive factor by altering its site of expression during excretory system development. Normally, GDNF is secreted by the metanephric mesenchyme and acts via receptors on the Wolffian duct and ureteric bud epithelium. Misexpression of GDNF in the Wolffian duct and ureteric buds resulted in formation of multiple, ectopic buds, which branched independently of the metanephric mesenchyme. This confirmed the ability of GDNF to induce ureter outgrowth and epithelial branching in vivo. However, in mutant mice lacking endogenous GDNF, kidney development was rescued to a substantial degree by GDNF supplied only by the Wolffian duct and ureteric bud. These results indicate that mesenchymal GDNF is not required as a chemoattractive factor to pattern the growth of the ureteric bud within the developing kidney, and that any positional information provided by the mesenchymal expression of GDNF may provide for renal branching morphogenesis is redundant with other signals. PMID- 15890331 TI - bloodthirsty, an RBCC/TRIM gene required for erythropoiesis in zebrafish. AB - The Antarctic icefishes (family Channichthyidae, suborder Notothenioidei) constitute the only vertebrate taxon that fails to produce red blood cells. These fishes can be paired with closely related, but erythrocyte-producing, notothenioids to discover erythropoietic genes via representational difference analysis. Using a B30.2-domain-encoding DNA probe so derived from the hematopoietic kidney (pronephros) of a red-blooded Antarctic rockcod, Notothenia coriiceps, we discovered a related, novel gene, bloodthirsty (bty), that encoded a 547-residue protein that contains sequential RING finger, B Box, coiled-coil, and B30.2 domains. bty mRNA was expressed by the pronephric kidney of N. coriiceps at a steady-state level 10-fold greater than that found in the kidney of the icefish Chaenocephalus aceratus. To test the function of bty, we cloned the orthologous zebrafish gene from a kidney cDNA library. Whole-mount in situ hybridization of zebrafish embryos showed that bty mRNA was present throughout development and, after the mid-blastula transition, was expressed in the head and in or near the site of primitive erythropoiesis in the tail just prior to red cell production. One- to four-cell embryos injected with two distinct antisense morpholino oligonucleotides (MOs) targeted to the 5'-end of the bty mRNA failed to develop red cells, whereas embryos injected with 4- and 5-bp mismatch control MOs produced wild-type quantities of erythrocytes. The morphant phenotype was rescued by co-injection of synthetic bty mRNA containing an artificial 5' untranslated region (UTR) with the antisense MO that bound the 5'-UTR of the wild type bty transcript. Furthermore, the expression of genes that mark terminal erythroid differentiation was greatly reduced in the antisense-MO-treated embryos. We conclude that bty is likely to play a role in differentiation of the committed red cell progenitor. PMID- 15890332 TI - Stage-specific changes in gene expression in acutely isolated mouse CNS progenitor cells. AB - Neural progenitor cells can be derived from a variety of developmental stages when they are preferentially proliferating, undergoing neurogenesis or undergoing gliogenesis. We used FACS sorting and the LeX surface marker to enrich neural progenitor cells from different embryonic stages and adult and compared their gene expression profiles using Affymetrix Microarrays. Our results show that, while there are common genes expressed in the progenitor cell population from all stages, there are also significant differences in gene expression patterns that correlate with stage-related behaviors. These data indicate that progenitor cells change during development and that adult and embryonic neural progenitor cells are intrinsically different. PMID- 15890333 TI - Transcription-dependent and transcription-independent functions of the classical progesterone receptor in Xenopus ovaries. AB - Two forms of the classical progesterone receptors (PR), XPR-1 and XPR-2, have been cloned in Xenopus laevis. Their relative roles in mediating progesterone action in the ovaries are not clear. Using antibodies generated against the cloned XPR-2, we demonstrated here that the somatic follicle cells expressed an 80-kDa PR protein, termed XPR-1. Treatment of follicle cells with progesterone resulted in disappearance of this protein, consistent with proteosome-mediated XPR-1 protein degradation. A smaller (approximately 70 kDa) PR protein, termed XPR-2, was expressed in the oocytes, but not in follicle cells. XPR-2 underwent progesterone-induced phosphorylation but not protein degradation. Treating isolated ovarian fragments with progesterone caused oocyte maturation and the release of the mature oocytes from the ovarian tissues ("ovulation"). Inhibition of transcription, with actinomycin D, did not interfere with progesterone-induced oocyte maturation but blocked "ovulation" so that mature oocytes were trapped in the follicles. These results suggest that the dual functions of progesterone, transcription-dependent follicle rupture and transcription-independent oocyte maturation, are mediated by the two forms of PR proteins differentially expressed in the follicle cells and the oocytes, respectively. PMID- 15890334 TI - SMA-1 spectrin has essential roles in epithelial cell sheet morphogenesis in C. elegans. AB - During Caenorhabditis elegans development, the embryo acquires its vermiform shape due to changes in the shape of epithelial cells, a process that requires an apically localized actin cytoskeleton. We show that SMA-1, an ortholog of beta(H) spectrin required for normal morphogenesis, localizes to the apical membrane of epithelial cells when these cells are rapidly elongating. In spc-1 alpha-spectrin mutants, SMA-1 localizes to the apical membrane but its organization is altered, consistent with the hypothesis these proteins act together to form an apically localized spectrin-based membrane skeleton (SBMS). SMA-1 is required to maintain the association between actin and the apical membrane; sma-1 mutant embryos fail to elongate because actin, which provides the driving force for cell shape change, dissociates from the apical membrane skeleton during morphogenesis. Analysis of sma-1 expression constructs and mutant strains indicates SMA-1 maintains the association between actin and the apical membrane via interactions at its N-terminus and this activity is independent of alpha-spectrin. SMA-1 also preserves dynamic changes in the organization of the apical membrane skeleton. Taken together, our results show the SMA-1 SBMS plays a dynamic role in converting changes in actin organization into changes in epithelial cell shape during C. elegans embryogenesis. PMID- 15890335 TI - Regulation of vertebrate myotome development by the p38 MAP kinase-MEF2 signaling pathway. AB - Biochemical and cell culture studies have characterized the myocyte enhancer factor 2 (MEF2) transcriptional regulatory proteins as obligatory partners for the myogenic regulatory factors (MRFs) in the differentiation of myogenic cells in culture. However, the role of MEF2 activation in somitic myogenesis has not been fully characterized. Here, we report a critical interaction between the p38 mitogen-activated protein kinase (p38 MAPK) and MEF2 in the developing somite myotome. We document expression of MEF2A and p38 MAPK proteins in the somite of 9.5 dpc mouse embryos concurrent with Myf 5 protein expression. We also observed that abrogation of p38 MAPK signaling blocks MEF2 activation using a MEF2 transgenic 'sensor' mouse. Inhibition of p38 MAPK signaling concurrently inhibited myogenic differentiation in somite cultures and in embryos in vivo using transplacental injection of a p38 inhibitor (SB203580). Finally, we document that commitment to the myogenic lineage is not appreciably affected by p38 MAPK inhibition since the activation of an early marker of myogenic commitment (Myf 5) occurs normally when p38 MAPK signaling is inhibited. Thus, we present novel evidence indicating a crucial role for p38 MAPK signaling to the MEF2 transcriptional regulators during early mammalian somite development and myotome formation. PMID- 15890336 TI - Comparison of the gastroprokinetic effects of ghrelin, GHRP-6 and motilin in rats in vivo and in vitro. AB - Ghrelin and motilin form a new family of structurally related peptides. We compared the gastroprokinetic effects of ghrelin, the ghrelin receptor agonist, growth hormone releasing peptide 6 (GHRP-6), and motilin in rats in vivo and in vitro. METHODS: Ghrelin, GHRP-6 or motilin (10-150 microg/kg) were injected i.p. and the effects on gastric emptying and transit were measured after intragastric application of Evans blue. In antral and fundic strips the effect of motilin, ghrelin or GHRP-6 was studied during electrical field stimulation (EFS) in the absence and presence of N(G)-nitro-l-arginine methyl ester hydrochloride (l-NAME) (300 microM). RESULTS: Ghrelin and GHRP-6 but not motilin accelerated gastric emptying and transit in rats. Ghrelin was more potent than GHRP-6 and the dose response relationship for ghrelin but not for GHRP-6 was bell-shaped. In fundic or antral strips, neural responses to EFS consisted of an on-relaxation that was reversed into a cholinergically mediated contraction by addition of the nitric oxide (NO)-synthase blocker, l-NAME. The post-stimulus off-contraction was cholinergically mediated. Under normal conditions, the ghrelin agonists reduced the on-relaxations in fundic strips and increased the cholinergic off contractions in antral and fundic strips. The concentration response curves in muscle strips of the fundus were bell-shaped with maximal effects for ghrelin at 1.2 microM (on-responses) and 0.66 microM (off-responses) and for GHRP-6 at 0.50 microM (on-responses) and 0.26 microM (off-responses). No effects were observed with motilin between 1 nM and 0.1 microM. Studies in the presence of l-NAME confirmed the effect of the ghrelin agonists on cholinergic excitatory motor responses. No effects were observed with motilin under the different experimental conditions. The presence of growth hormone secretagogue receptor 1a transcripts in the strip preparations was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). CONCLUSION: Ghrelin and GHRP-6 but not motilin accelerate gastric emptying and transit by activating cholinergic excitatory pathways in the enteric nervous system in addition to the known vagal pathways. PMID- 15890337 TI - CSK negatively regulates nerve growth factor induced neural differentiation and augments AKT kinase activity. AB - Src family kinases are involved in transducing growth factor signals for cellular differentiation and proliferation in a variety of cell types. The activity of all Src family kinases (SFKs) is controlled by phosphorylation at their C-terminal 527-tyrosine residue by C-terminal SRC kinase, CSK. There is a paucity of information regarding the role of CSK and/or specific Src family kinases in neuronal differentiation. Pretreatment of PC12 cells with the Src family kinase inhibitor, PP1, blocked NGF-induced activation of SFKs and obliterated neurite outgrowth. To confirm a role for CSK and specific isoforms of SFKs in neuronal differentiation, we overexpressed active and catalytically dead CSK in the rat pheochromocytoma cell line, PC12. CSK overexpression caused a profound inhibition of NGF-induced activation of FYN, YES, RAS, and ERK and inhibited neurite outgrowth, NGF-stimulated integrin-directed migration and blocked the NGF-induced conversion of GDP-RAC to its GTP-bound active state. CSK overexpression markedly augmented the activation state of AKT following NGF stimulation. In contrast, kinase-dead CSK augmented the activation of FYN, RAS, and ERK and increased neurite outgrowth. These data suggest a distinct requirement for CSK in the regulation of NGF/TrkA activation of RAS, RAC, ERK, and AKT via the differential control of SFKs in the orchestration of neuronal differentiation. PMID- 15890338 TI - Repeated 2 Hz peripheral electrical stimulations suppress morphine-induced CPP and improve spatial memory ability in rats. AB - Our previous studies have shown that 2 Hz peripheral electrical stimulation (PES) can suppress morphine-induced conditioned place preference (CPP) in the rat, although the mechanisms remain unclear. Since CPP involves the mechanism of learning and memory, it is rational to ask whether the suppressive effect of repeated 2 Hz PES on morphine-induced CPP is due to an impairment of the function of spatial learning and memory. Rats were trained with 4 mg/kg morphine, i.p. for 4 days to establish the CPP. Twenty-four hours after the CPP testing, they were given PES at 2 Hz once a day for 1, 3 or 5 days, followed by another CPP testing. The results showed that (1) the morphine-induced CPP was significantly inhibited by 3 or 5 consecutive sessions, but not by single session of 2 Hz PES. (2) A test of spatial leaning and memory ability using the Morris water maze task revealed that 2 Hz PES per se exhibited a promoting, rather than a deteriorating effect on the ability of spatial memory. (3) 2 Hz PES by itself produced a moderate yet significant CPP. The results imply that (a) a low frequency PES can produce a rewarding effect as revealed by the CPP testing, which may account, at least in part, for its suppressive effect on morphine induced CPP, (b) the suppressive effect of PES on morphine induced CPP is not due to a deteriorating effect on the ability of spatial memory. PMID- 15890339 TI - Schwann cell behavior in three-dimensional collagen gels: evidence for differential mechano-transduction and the influence of TGF-beta 1 in morphological polarization and differentiation. AB - Schwann cells (SCs) cultured on and within magnetically aligned collagen gels were examined for their abilities to spread and exhibit contact guidance, two functions that are relevant to their potential enhancement of neurite migration and regeneration in entubulation repair of transection-type nerve injuries. Cells seeded at or near the surfaces of gels abandoned their initially spherical shapes, adopting spread morphologies rapidly compared to cells within the gels. Those few cells within the gels that did spread exhibited marked contact guidance responses, aligning strongly with the aligned collagen fibrils. Spreading of cells in gels could not be induced by varied cell concentration, collagen density, mitogen presence, inclusion of soluble laminin, or use of fibrin gel in lieu of collagen. However, cells that settled at the interface between collagen gel layers during gellation of the top layer above a preformed bottom layer were highly spread. This suggests that a differential mechanical interaction across the cell at an interface, where at least one surface presents constituents of the basal lamina, permits the Schwann cell to rapidly revert to a spread, differentiated phenotype. Unlike other reagents, TGF-beta1 was able to induce significant SC spreading as early as 4 h post-seeding. Consistent with the differential-mechanical cue mechanism, TGF-beta1 appears to facilitate this response, at least in part, by upregulating beta1 integrin expression, thereby enabling the SC to more acutely detect these local cues in the mechanical environment. PMID- 15890340 TI - Comparison of effects of methylprednisolone and anti-CD11d antibody treatments on autonomic dysreflexia after spinal cord injury. AB - Autonomic dysreflexia is a condition of episodic hypertension that develops after spinal cord injury (SCI). We previously showed that a two-day anti-inflammatory treatment with an anti-CD11d integrin monoclonal antibody (mAb), soon after SCI in rats, reduced the magnitude of dysreflexia for at least 6 weeks. Effects of methylprednisolone (MP), a commonly used neuroprotective treatment for SCI, on dysreflexia have never been examined. We compared the effects of a 2-day MP treatment and/or the anti-CD11d mAb on autonomic dysreflexia, elicited by colon distension, after clip-compression SCI at the 4th thoracic segment (T4) in rats. We assessed the effects of each treatment on the size of the calcitonin gene related peptide (CGRP)-immunoreactive afferent arbour in the dorsal horn, as changes in this arbour can correlate with the development of dysreflexia. MP reduced autonomic dysreflexia by approximately 50% at 2 weeks after SCI, but this effect was lost by 6 weeks. At 2 weeks, the combined effects of MP and the mAb were not additive, reducing dysreflexia by approximately 50%. Neither MP nor the mAb treatment altered the area of CGRP-immunoreactive fibres in the lumbar cord, the crucial input region for dysreflexia initiated by colon distension. However, both treatments led to increased fibre areas in the T9 segment, correlated with greater tissue integrity and smaller lesions, delineated by inflammatory cells. In summary, MP only temporarily decreases autonomic dysreflexia after SCI. The early beneficial effects of both treatments on dysreflexia do not relate to changes in the CGRP-immunoreactive afferent arbour but may correlate with decreased lesion progression. PMID- 15890341 TI - Proteasome plasticity. AB - The 26S proteasome is responsible for regulated proteolysis of most intracellular proteins yet the focus of intense regulatory action itself. Proteasome abundance is responsive to cell needs or stress conditions, and dynamically localized to concentrations of substrates. Proteasomes are continually assembled and disassembled, and their subunits subject to a variety of posttranslational modifications. Furthermore, as robust and multi-tasking as this complex is, it does not function alone. A spattering of closely associating proteins enhances complex stability, fine-tunes activity, assists in substrate-binding, recycling of ubiquitin, and more. HEAT repeat caps activate proteasomes, yet share remarkable features with nuclear importins. Fascinating cross talk even occurs with ribosomes through common maturation factors. The dynamics of proteasome configurations and how they relate to diverse activities is the topic of this review. PMID- 15890342 TI - Thermodynamics of phosphorylcholine and lysophosphatidylcholine binding to the major protein of bovine seminal plasma, PDC-109. AB - PDC-109 binds to sperm plasma membranes by specific interaction with choline phospholipids and induces cholesterol efflux, a necessary event before capacitation - and subsequent fertilization - can occur. The binding of phosphorylcholine (PrC) and lysophosphatidylcholine (Lyso-PC) with PDC-109 was investigated by monitoring the ligand-induced changes in the absorption spectrum of PDC-109. At 20 degrees C, the association constants (K(a)), for PrC and Lyso PC were obtained as 81.4M(-1) and 2.02 x 10(4) M(-1), respectively, indicating that the binding of Lyso-PC to PDC-109 is 250-fold stronger than that of PrC. From the temperature dependence of the K(a) values, enthalpy of binding (DeltaH(0)) and entropy of binding (DeltaS(0)), were obtained as -79.7 and -237.1 J mol(-1)K(-1) for PrC and -73.0 kJ mol(-1) and -167.3 J mol(-1)K(-1) for Lyso PC, respectively. These results demonstrate that although the binding of these two ligands is driven by enthalpic forces, smaller negative entropy of binding associated with Lyso-PC results in its significantly stronger binding. PMID- 15890343 TI - Functional analysis of transcriptional repressor Otx3/Dmbx1. AB - Otx3/Dmbx1 is a member of paired class homeodomain transcription factors. In this study, we found that Otx3/Dmbx1 represses the Otx2-mediated transactivation by forming heterodimer with Otx2 on the P3C (TAATCCGATTA) sequence in vitro. The 156 amino acid region (residues 1-156) of Otx3/Dmbx1 is required for its repressor activity, and interacts directly with Otx2. Co-localization of Otx3/Dmbx1 and Otx2 in brain sections was confirmed by in situ hybridization. These data suggest that Otx3/Dmbx1 represses Otx2-mediated transcription in the developing brain. We also identified the consensus binding sequence [TAATCCGATTA and TAATCC(N2 4)TAATCC] of Otx3/Dmbx1. PMID- 15890344 TI - Induction of ornithine decarboxylase in T/C-28a2 chondrocytes by lysophosphatidic acid: signaling pathway and inhibition of cell proliferation. AB - Among several extracellular messengers tested, lysophosphatidic acid (LPA) was able to cause the most marked induction of ornithine decarboxylase (ODC) in serum starved human T/C-28a2 chondrocytes. LPA also induced the activation/phosphorylation of Src, Akt and p44/42 MAPK, and the translocation of PKC-delta from cytosol to membrane coupled to its tyrosine phosphorylation. Experiments with selective signaling inhibitors indicate that LPA leads to Src activation through Gi protein-coupled receptors. In turn Src can activate PI3K and PKC-delta, and all these signaling proteins are required for ODC induction. In conclusion these results show that chondrocytes may be a novel target for LPA action. However, although LPA is considered a mitogen for several cell types and ODC induction is generally correlated to cell growth, LPA was not able to stimulate chondrocyte growth, but rather exerted an anti-proliferative effect. PMID- 15890345 TI - Abeta(31-35) and Abeta(25-35) fragments of amyloid beta-protein induce cellular death through apoptotic signals: Role of the redox state of methionine-35. AB - In order to clarify the basis of neuronal toxicity exerted by the shortest active peptides of amyloid beta-protein (Abeta), the toxic effects of Abeta(31-35) and Abeta(25-35) peptides on isolated rat brain mitochondria were investigated. The results show that exposure of isolated rat brain mitochondria to Abeta(31-35) and Abeta(25-35) peptides determines: (i) release of cytochrome c; (ii) mitochondrial swelling and (iii) a significant reduction in mitochondrial oxygen consumption. In contrast, the amplitude of these events resulted attenuated in isolated brain mitochondria exposed to the Abeta(31-35)Met35(OX) in which methionine-35 was oxidized to methionine sulfoxide. The Abeta peptide derivative with norleucine substituting Met-35, i.e., Abeta(31-35)Nle-35, had not effect on any of the biochemical parameters tested. We have further characterized the action of Abeta(31-35) and Abeta(25-35) peptides on neuronal cells. Taken together our result indicate that Abeta(31-35) and Abeta(25-35) peptides in non-aggregated form, i.e., predominantly monomeric, are strongly neurotoxic, having the ability to enter within the cells, determining mitochondrial damage with an evident trigger of apoptotic signals. Such a mechanism of toxicity seems to be dependent by the redox state of methionine-35. PMID- 15890346 TI - Blocking the cleavage at midportion between gamma- and epsilon-sites remarkably suppresses the generation of amyloid beta-protein. AB - To examine how gamma- and epsilon-cleavages of beta-amyloid precursor protein (APP) are related, each cleavage site was replaced with a stretch of Trp that cannot be cleaved by gamma-secretase. Replacement of the gamma- or epsilon-site significantly suppressed secretion of amyloid beta-protein (Abeta), and produced longer Abeta or longer APP intracellular domain, respectively. This cleavage at the midportion between gamma- and epsilon-sites was also gamma-secretase dependent. Blocking this cleavage with a Trp stretch remarkably suppressed Abeta generation, indicating that the midportion cleavage is required for the generation of Abeta. PMID- 15890347 TI - ICAM-3, a ligand for DC-SIGN, was duplicated from ICAM-1 in mammalian evolution, but was lost in the rodent genome. AB - ICAM-3 is a DC-SIGN ligand that is constitutively expressed on resting leukocytes, and is thus an important molecule for the first immune response. But, ICAM-3 has not been isolated form rodents. Thus, we compare the ICAM gene clusters in human, dog, mouse, and rat. ICAM-1, -4, -5 and -3 are located close to one another on the same chromosome and show genomic synteny in human and dog. Almost the same ICAM gene clusters were found in rodent genome, but only the ICAM 3 was not present. A phylogenetic tree plotting the cDNAs of human, dog, mouse, rat, and bovine suggested that ICAM-3 was made from a duplication of ICAM-1. Thus, ICAM-3 arose from ICAM-1 in the mammalian evolution, but was lost in the rodent's genome. Our study suggests the different immune response in the rodents in comparison with other mammals. PMID- 15890348 TI - Spotlight on... Irmgard Sinning. PMID- 15890349 TI - Conductimetric determination of phenylpropanolamine HCl, ranitidine HCl, hyoscyamine HBr and betaine HCl in their pure state and pharmaceutical preparations. AB - Sodium tetraphenylborate and phosphotungstic acid were used as titrants for the conductimetric determination of phenylpropanolamine HCl (PPA.Cl), ranitidine HCl (Ra.Cl), hyoscyamine HBr (Hy.Br) and betaine HCl (Be.Cl) through ion-associate complex formation. The molar combining ratio and the solubility products of the formed ion-associates were studied and calculated. The suggested method has been applied to the determination of the mentioned drugs in their pure state and pharmaceutical preparations with mean recovery values of 97.71-102.97% and relative standard deviations 0.25-0.85%. The accuracy of the method is indicated by excellent recovery and low standard deviation. The results are compared with the pharmacopoeial or the official methods. PMID- 15890350 TI - Testosterone and estradiol produce different effects on cognitive performance in male rats. AB - The effects of castration and hormone treatment on cognitive performance were evaluated in male rats. Castrated animals received either testosterone or estradiol and were compared with gonadally intact animals and with castrated controls. Results revealed a dissociation between the effects of testosterone and estradiol on cognitive performance in male rats. Specifically, estradiol enhanced acquisition of a delayed matching-to-position spatial task, similar to previously published observations in females. In contrast, neither castration nor testosterone treatment had any significant effect on acquisition of the delayed matching-to-position task, but did appear to affect delay-dependent working memory. None of the treatments had any significant effect on acquisition of a configural association negative patterning task, suggesting that effects on the delayed matching-to-position task were not due to effects on motivational factors. These data demonstrate that, as in females, gonadal hormones influence cognitive performance in males and suggest that estradiol and testosterone affect distinct cognitive domains. PMID- 15890351 TI - Efficacy of a DNA vaccine delivered in attenuated Salmonella typhimurium against Eimeria tenella infection in chickens. AB - The efficacy of an oral DNA vaccine carrying the Eimeria tenella 5401 antigen gene delivered by attenuated Salmonella typhimurium was examined in an experimental challenge study. The DNA vaccine preparation was made by transforming the recombinant plasmid pcDNA3-5401 into the attenuated S. typhimurium strain (Dam(-) and PhoP(-)) (designated hereafter as ZJ111/pcDNA3 5401). The chickens were randomly divided into six groups, 50 per group. Group A were given PBS as control. Chickens in group B were fed with 10(8) colony forming units (CFU) of attenuated S. typhimurium carrying pcDNA3. Group C were immunised with 100 microg of the recombinant 5401 protein via intramuscular injection. Groups D to F orally received ZJ111/pcDNA3-5401 at doses of 10(7), 10(8) and 10(9)CFU per chicken, respectively. All immunisations were boosted 2 weeks later. The immunised chickens were challenged with 6x10(4) homologous sporulated oocysts 14 days after the second immunisation. No significant differences in body weight were detected between the groups before immunisation and at week 4 after the booster immunisation. The ZJ111/pcDNA3-5401 was eventually eliminated from the spleen and liver on week 6 post-immunisation. The plasmid pcDNA3-5401 was stably maintained in over 80% of the attenuated S. typhimurium population after 100 generations of growth in antibiotic-free media. Oral immunisation of chickens with ZJ111/pcDNA3-5401 elicited specific humoral responses and stimulated proliferation of peripheral blood lymphocytes. The lymphocyte proliferation response was significantly higher in all vaccinated groups than in the control chickens. Antibody response was significantly lower in group C than in groups immunised with strain ZJ111/pcDNA3-5401. Vaccination with the strain ZJ111/pcDNA3 5401 at 10(8) (group E) and 10(9) (group F) CFU per chicken provided 55.0 and 57.5% protection against E. tenella challenge, respectively. These results have important implications for the development of DNA vaccines against avian coccidiosis by bacteria-vectored oral delivery system. PMID- 15890352 TI - Psychophysical design influences frequency discrimination performance in young children. AB - We investigated how different psychophysical procedures affect frequency discrimination performance in children. Four studies used a design in which listeners heard two tone pairs and had to identify whether the first or second pair contained a higher frequency target tone. Thresholds for 6- and 7-year-olds were higher than those for 8- and 9-year-olds and adults. Two manipulations led to lower (better) thresholds in young children: (a) moving the standard comparison tone before the target tone and (b) using three target comparison tone pairs. It is suggested that young children benefit from designs that help cue them to when they need to attend to a target tone. The two-interval, forced choice procedure that is widely used in studies of developmental disorders led to variable performance even in adults and did not give a realistic picture of the perceptual capabilities of children under 8 years of age. PMID- 15890353 TI - Detection of circulating autoantibodies directed against the asialoglycoprotein receptor using recombinant receptor subunit H1. AB - BACKGROUND/AIMS: The asialoglycoprotein-receptor (ASGPR) is a major liver specific target autoantigen in autoimmune hepatitis. ASGPR heteromers of two subunits H1 and H2 provide clearance of circulating asialoglycoproteins by receptor-mediated endocytosis. The aim of this study was to establish whether a recombinantly expressed subunit H1 presenting conformational epitopes is capable of detecting autoantibodies against ASGPR in patients with inflammatory liver disease. METHODS: The major subunit H1 was expressed in human-embryo-kidney 293 cells and prepared by ligand-affinity-chromatography similar to the complete receptor from normal liver. Reactivities of anti-ASGPR positive sera from 219 patients with both recombinant H1 and natural receptor were compared using enzyme immunoassay (EIA). RESULTS: 194 of 219 sera (88.6%) showed absorbance values on 293-H1 within a range of +/-15% compared to the natural receptor. 145 of 149 sera (97.3%) positive on ASGPR were also positive on recombinant H1. Titers of 61/62 sera (98.4%) revealed no deviation of more than one dilution step. ASGPR reactivity could be inhibited in 29 sera with up to 50 ng/microl of 293-H1. CONCLUSIONS: These results indicate that the antigenic sites of the human ASGPR are mainly located on the mammalian-expressed subunit H1. Therefore, 293-H1 is a powerful tool for the detection of autoantibodies against ASGPR. PMID- 15890354 TI - Cryopreserved dendritic cells for intratumoral immunotherapy do not require re culture prior to human vaccination. AB - Dendritic cell (DC) immunotherapy for cancer has shown great promise so far. The ability to deliver dendritic cells directly into tumours where they are capable of acquiring tumour antigens prior to stimulating specific T cell responses has been demonstrated both in animal models and human patients. Clinical grade DCs can be grown from peripheral blood monocytes in the absence of foetal calf serum (FCS) and cryopreserved to generate plentiful identical aliquots thus avoiding repeated venesection. However, the approach is still limited by the necessity to return thawed DCs to culture prior to injection. It would be more advantageous to directly inject the DCs whilst still in the freezing medium and thus prevent the need for further manipulation. Whilst several reports have shown that cryopreserved DCs can survive for over 72 h when returned to culture, there is no information regarding the longevity of cells maintained in the freezing medium after thawing. In this report we have shown that DCs may remain in freezing medium for up to 1 h without affecting their survival, phenotype or function. This period of time is sufficient to allow for any delays incurred between the preparation of the DCs and time taken to be administered within a standard clinical setting. This study demonstrates that clinical grade DCs can be cryopreserved and thawed whilst retaining the ability to acquire exogenous antigenic material required for intratumoural immunotherapy. The survival of these cells within the freezing medium without the requirement for re-culture expands their availability for administration directly to the tumours of patients in non-specialist centres that do not have the appropriate facilities for DC re culture. PMID- 15890355 TI - IgG antibodies against bovine serum albumin in humans--their prevalence and response to exposure to bovine serum albumin. AB - Human exposure to bovine serum albumin (BSA) is very common and occurs through dietary and medicinal routes. Although great effort has been made to reduce exposure to BSA in pharmaceuticals to eliminate the threat of bovine spongiform encephalopathy, less attention has been given to assessing the human immune response after exposure to BSA. A sensitive quantitative radioimmunoassay was therefore developed to measure anti-BSA IgG antibodies in healthy subjects and in cancer patients participating in a randomized, placebo controlled clinical trial where they were exposed to BSA as an intrathoracic surgical sealant during pneumonectomy. Anti-BSA antibodies were detected in 55% of 60 healthy blood donors and 51% of 83 patients before lung cancer resection. The median antibody levels were the same in both cohorts; 0.086 microg/mL (range 0.016-19.5 microg/mL) for health blood donors and 0.062 microg/mL (range 0.009-44 microg/mL) for cancer patients. Six months after exposure of the cancer patients to BSA, the percentage of patients with anti-BSA antibody rose to 96% and the median antibody level rose to 19 microg/mL (range 0.009-258 microg/mL). Placebo-treated cancer patients showed no significant increase in the percentage of patients with anti BSA antibody (41%) or the median antibody level (0.047 microg/mL; range 0.008 1.58) over 6 months. Western blot analysis confirmed the presence of anti-BSA antibody. Elevated levels of anti-BSA antibody were not associated with any detectable clinical events in either the healthy blood donors or the cancer patients. PMID- 15890356 TI - Morphological and enzymatic analysis of the midgut of Anopheles darlingi during blood digestion. AB - The midgut of adult female Anopheles darlingi is comprised of narrow anterior and dilated posterior regions, with a single layered epithelium composed by cuboidal digestive cells. Densely packed apical microvilli and an intricate basal labyrinth characterize each cell pole. Before blood feeding, apical cytoplasm contains numerous round granules and whorled profiles of rough endoplasmic reticulum. Engorgement causes a great distension of midgut. This provokes the flattening of digestive cells and their nuclei. Simultaneously, apical granules disappear, the whorls of endoplasmic reticulum disassemble and 3h post bloodmeal (PBM), nucleoli enlarge manyfold. An intense absorptive process takes place during the first 24 h PBM, with the formation of large glycogen inclusions, which persist after the end of the digestive process. Endoproteases activities are induced after bloodmeal and attain their maximum values between 10 and 36 h PBM. At least two different aminopeptidases seem to participate in the digestive process, with their maximum activity values at 36 and 48 h PBM, respectively. Coarse electrondense aggregates, possibly debris from digested erythrocytes, begin to appear on the luminal face of the peritrophic membrane from 18 h PBM and persist during all the digestive process, and are excreted at its end. We suggest that these aggregates could contain some kind of insoluble form of haem, in order of neutralize its toxicity. PMID- 15890357 TI - The gp130 ligand oncostatin M regulates tissue inhibitor of metalloproteinases-1 through ERK1/2 and p38 in human adult cardiac myocytes and in human adult cardiac fibroblasts: a possible role for the gp130/gp130 ligand system in the modulation of extracellular matrix degradation in the human heart. AB - There is ample evidence supporting the view that alterations in the balance between matrix deposition and matrix degradation brought about by changes in the respective activities of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) contribute significantly to cardiac dysfunction and disease. Here we show that TIMP-1 was upregulated up to threefold after treatment with the inflammatory mediator and gp130 ligand oncostatin M (OSM) in human adult cardiac myocytes and fibroblasts. The Erk1/2 inhibitor PD98059 and the p38 inhibitor SD202190 abolished the effect of OSM on TIMP-1 production in both cell types. Human cardiac myocytes and human cardiac fibroblasts also express MMP-1, 2, 3 and 9, and TIMP-2 constitutively. OSM, however, did not affect the expression of these proteins. In addition also the other gp130 ligands tested, cardiotrophin-1 (CT-1), interleukin-6 (IL-6) and leukemia inhibitory factor (LIF) had no effect on the expression of TIMPs and MMPs studied. We speculate that OSM by inducing TIMP-1 expression counteracts excessive proteolysis and unrestricted matrix degradation during inflammatory processes in the heart. The notion that OSM favors matrix stabilization in the human heart is further supported by our earlier observation that OSM also upregulates PAI-1, the physiological inhibitor of the protease urokinase-type PA (u-PA), which in turn is essential for extracellular proteolysis. Therefore we propose a role for the gp130 ligand OSM in the modulation of cardiac remodeling and repair processes. PMID- 15890358 TI - L-Thyroxine activates Akt signaling in the heart. AB - Hyperthyroidism causes physiological cardiac hypertrophy and enhanced function. Many of these effects have been traditionally attributed to changes in gene expression. However, the role of signal transduction pathways in the effects mediated by thyroid hormone (TH) have recently gained a significant amount of attention in non-cardiovascular cells and tissue. Whether signal transduction pathways are involved in the cardiac effects of TH is unknown. In this study, we treated Sprague Dawley rats with L-thyroxine (T4) or propylthiouracil (PTU) to determine whether there was modulation of signal transduction pathways in the left ventricle. Predictably, T4 increased heart weight, left ventricular systolic pressure, and dP/dT. T4 and PTU also had typical effects on expression of thyroid responsive genes such as alpha and beta myosin heavy chain. T4 treatment caused phosphorylation of Akt and downstream signaling components such as GSK-3beta, mTOR, and S6 kinase. In conclusion, activation of the Akt signaling pathway may contribute to the effects of TH on the heart. While this pathway is clearly activated, further work is needed to determine whether this is via a genomic or non-genomic mechanism. PMID- 15890359 TI - Llama single domain antibodies as a tool for molecular mimicry. AB - In camelids, a subset of the immunoglobulins consists of heavy-chain homodimers devoid of light chains, and are thus called heavy-chain IgGs (hcIgGs). Their variable region (VHH) is the smallest antigen-binding fragment possible, and being just one polypeptide chain it is especially suitable for engineering. In particular, camelid single domain antibodies might be very useful for molecular mimicry and anti-idiotypic vaccination. In the present work, we show that llamas immunized with an anti-DNA mouse mAb develop an important anti-Id response. Selection of VHHs by phage display, with specific elution of bound phages with the external antigenic DNA, shows that selected private anti-Id VHHs compete for binding to the external antigen and bear a functional mimicry of the DNA. These results indicate that llama anti-Id single domain antibodies would be an excellent tool for molecular mimicry studies. PMID- 15890360 TI - Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. AB - The cyclin-dependent kinase inhibitors (CKIs) bind to and directly regulate the catalytic activity of cyclin-dependent kinase (Cdk)/cyclin complexes involved in cell cycle control and do not regulate other, closely related Cdks. We showed previously that the CKI, p27, binds to Cdk2/cyclin A though a sequential mechanism that involves folding-on-binding. The first step in the kinetic mechanism is interaction of a small, highly dynamic domain of p27 (domain 1) with the cyclin subunit of the Cdk2/cyclin A complex, followed by much slower binding of a more lengthy and less flexible domain (domain 2) to Cdk2. The second step requires folding of domain 2 into the kinase inhibitory conformation. Rapid binding of p27 domain 1 to cyclin A tethers the inhibitor to the binary Cdk2/cyclin A complex, which reduces the entropic barrier associated with slow binding of domain 2 to the catalytic subunit. We show here that p27/cyclin interactions are an important determinant of p27 specificity towards cell cycle Cdks. We used surface plasmon resonance, limited proteolysis, mass spectrometry, and NMR spectroscopy to study the interaction of p27 with Cdk2/cyclin A, and with another Cdk complex, Cdk5/p25, that is involved in neurodegeneration. Importantly, Cdk5/p35 (the parent complex of Cdk5/p25) is not regulated by p27 in neurons. Our results show that p27 binds to Cdk5 and Cdk2 with similar, slow kinetics. However, p27 fails to interact with p25 within the Cdk5/p25 complex, which we believe prevents formation of a kinetically trapped, inhibited p27/Cdk5/p25 complex in vivo. The helical topology of p25 is very similar to that of cyclin A. However, p25 lacks the MRAIL sequence in one helix that, in the cell cycle cyclins, mediates specific interactions with domain 1 of p21 and p27. Our results strongly suggest that p21 and p27, related Cdk inhibitors, select their cell cycle regulatory Cdk targets by binding specifically to the cyclin subunit of these Cdk/cyclin complexes as a first step in a sequential, folding-on-binding mechanism. PMID- 15890361 TI - 13C NMR relaxation studies of RNA base and ribose nuclei reveal a complex pattern of motions in the RNA binding site for human U1A protein. AB - The widespread importance of induced fit and order-disorder transition in RNA recognition by proteins and small molecules makes it imperative that RNA motional properties are characterized quantitatively. Until now, however, very few studies have been dedicated to the systematic characterization of RNA motion and to their changes upon protein or small-molecule binding. The U1A protein-RNA complexes provide some of the best-studied examples of the role of RNA motional changes upon protein binding. Here, we report (13)C NMR relaxation studies of base and ribose dynamics for the RNA internal loop target of human U1A protein located within the 3'-untranslated region (3'-UTR) of the mRNA coding for U1A itself. We also report the semi-quantitative analysis of both fast (nano- to picosecond) and intermediate (micro- to millisecond) motions for this paradigmatic RNA system. We measure (13)C T(1), T(1rho) and heteronuclear nuclear Overhauser effects (NOEs) for sugar and base nuclei, as well as the power dependence of T(1rho) at 500 MHz and 750 MHz, and analyze these results using the model-free formalism. The results provide a much clearer picture of the type of motions experienced by this RNA in the absence of the protein than was provided by the analysis of the structure based solely on NOEs and scalar couplings. They define a model where the RNA internal loop region "breathes" on a micro- to millisecond timescale with respect to the double-helical regions. Superimposed on this slower motion, the residues at the very tip of the loop undergo faster (nano- to picosecond) motions. We hypothesize that these motions allow the RNA to sample multiple conformations so that the protein can select a structure within the ensemble that optimizes intermolecular contacts. PMID- 15890362 TI - Direct characterization of the folded, unfolded and urea-denatured states of the C-terminal domain of the ribosomal protein L9. AB - The stability of the isolated C-terminal domain of the ribosomal protein L9 (CTL9) is strongly dependent upon pH. Below pH 4.2, the folded and unfolded states are both populated significantly. Their interconversion is slow on the NMR chemical shift time-scale and separate, well-resolved resonances from each state are observed. This allows the hydrodynamic properties of both states to be studied under identical conditions by using pulse field gradient NMR experiments. Hydrodynamic radii of the folded, unfolded and urea denatured protein molecules at pD 3.8 have been derived. The acid-denatured protein has a significantly smaller hydrodynamic radius, 28.2A, compared to that of the urea-denatured protein, which is 33.6A at pD 3.8. Far-UV CD spectra show that there is more residual secondary structure retained in the acid-denatured ensemble than in the urea-denatured one. ANS binding experiments and analysis of the CD data show that this acid-denatured species is not a molten globule state. Diffusion measurements of CTL9 were conducted over the pD range from 2.1 to 7.0. The hydrodynamic radii of both the folded and the acid-unfolded protein start to increase below pD 4, with the radius of hydration of the acid-unfolded state increasing from 25.1A at pD 4.2 to 33.5A at pD 2.1. The hydrodynamic radius of the urea-denatured protein is much less sensitive to pH. The unfolded protein at pD 2.1, no urea, has almost the same hydrodynamic radius as the urea-denatured protein at pD 3.8. The CD spectra, however, show significant differences in residual secondary structure, and the acid-denatured state contains more structure. PMID- 15890363 TI - A single residue of GDF-5 defines binding specificity to BMP receptor IB. AB - Growth and differentiation factor 5 (GDF-5), a member of the TGF-beta superfamily, is involved in many developmental processes, like chondrogenesis and joint formation. Mutations in GDF-5 lead to diseases, e.g. chondrodysplasias like Hunter-Thompson, Grebe and DuPan syndromes and brachydactyly. Similar to other TGF-beta superfamily members, GDF-5 transmits signals through binding to two different types of membrane-bound serine-/threonine-kinase receptors termed type I and type II. In contrast to the large number of ligands, only seven type I and five type II receptors have been identified to date, implicating a limited promiscuity in ligand-receptor interaction. However, in contrast to other members of the TGF-beta superfamily, GDF-5 shows a pronounced specificity in type I receptor interaction in cross-link experiments binding only to BMP receptor IB (BMPR-IB). In mice, deletion of either GDF-5 or BMPR-IB results in a similar phenotype, indicating that GDF-5 signaling is highly dependent on BMPR-IB. Here, we demonstrate by biosensor analysis that GDF-5 also binds to BMP receptor IA (BMPR-IA) but with approximately 12-fold lower affinity. Structural and mutational analyses revealed a single residue of GDF-5, Arg57 located in the pre helix loop, being solely responsible for the high binding specificity to BMPR-IB. In contrast to wild-type GDF-5, variant GDF-5R57A interacts with BMPR-IA and BMPR IB with a comparable high binding affinity. These results provide important insights into how receptor-binding specificity is generated at the molecular level and might be useful for the generation of receptor subtype specific activators or inhibitors. PMID- 15890364 TI - SCFGrr1-mediated ubiquitination of Gis4 modulates glucose response in yeast. AB - The F box protein Grr1 is the substrate specificity-determinant of the SCF(Grr1) E3 ubiquitin ligase complex. Genetic analyses of Grr1 mutants have implicated Grr1 in glucose repression, specifically with regard to expression of the SUC2 transcript. To better understand Grr1, we screened for substrates using a mutant version of Grr1 that should not associate with the SCF complex. We identified Gis4 as a novel Grr1 substrate. Gis4 was originally isolated as a multi-copy suppressor of a Gal--phenotype in the triple mutant snf1 mig1 srb8. Here, we show that Gis4 binds Grr1 in vivo and that Grr1 protein levels positively affect the protein levels of Gis4. The Gis4 protein is stable in wild-type cells and in grr1Delta cells; however, Gis4 is ubiquitinated in a Grr1-dependent manner. Furthermore, we show that Gis4 interacts with Snf1 in a Grr1-dependent fashion, and that Gis4 is involved in de-repression of SUC2 and in transcription of other Snf1-dependent transcripts. Gis4 appears to connect the glucose repression and de repression pathways. We suggest that Gis4 may explain the glucose repression defects in carbon source metabolism for the grr1 mutants. PMID- 15890365 TI - Elucidation of the stator organization in the V-ATPase of Neurospora crassa. AB - V-ATPases are membrane protein complexes that pump protons in the lumen of various subcellular compartments at the expense of ATP. Proton pumping is done by a rotary mechanism that requires a static connection between the membrane pumping domain (V(0)) and the extrinsic catalytic head (V(1)). This static connection is composed of several known subunits of the V-ATPase, but their location and topological relationships are still a matter of controversy. Here, we propose a model for the V-ATPase of Neurospora crassa on the basis of single-particle analysis by electron microscopy. Comparison of the resulting map to that of the A ATPase from Thermus thermophilus allows the positioning of two subunits in the static connecting region that are unique to eukaryotic V-ATPases (C and H). These two subunits seem to be located on opposite sides of a semicircular arrangement of the peripheral connecting elements, suggesting a role in stabilizing the stator in V-ATPases. PMID- 15890366 TI - Dynamics and metal exchange properties of C4C4 RING domains from CNOT4 and the p44 subunit of TFIIH. AB - Zinc fingers are small structured protein domains that require the coordination of zinc for a stable tertiary fold. Together with FYVE and PHD, the RING domain forms a distinct class of zinc-binding domains, where two zinc ions are ligated in a cross-braced manner, with the first and third pairs of ligands coordinating one zinc ion, while the second and fourth pairs ligate the other zinc ion. To investigate the relationship between the stability and dynamic behaviour of the domains and the stability of the metal-binding site, we studied metal exchange for the C4C4 RING domains of CNOT4 and the p44 subunit of TFIIH. We found that Zn(2+)-Cd(2+) exchange is different between the two metal-binding sites in the C4C4 RING domains of the two proteins. In order to understand the origins of these distinct exchange rates, we studied the backbone dynamics of both domains in the presence of zinc and of cadmium by NMR spectroscopy. The differential stability of the two metal-binding sites in the RING domains, as reflected by the different metal exchange rates, can be explained by a combination of accessibility and an electrostatic ion interaction model. A greater backbone flexibility for the p44 RING domain as compared to CNOT4 may be related to the distinct types of protein-protein interactions in which the two C4C4 RING domains are involved. PMID- 15890367 TI - CryoEM structure at 9A resolution of an adenovirus vector targeted to hematopoietic cells. AB - We report a sub-nanometer resolution cryo-electron microscopy (cryoEM) structural analysis of an adenoviral vector, Ad35F, comprised of an adenovirus type 5 (Ad5) capsid pseudo-typed with an Ad35 fiber. This vector transduces human hematopoietic cells via association of its fiber protein with CD46, a member of the complement regulatory protein family. Major advances in data acquisition and image processing allowed a significant improvement in resolution compared to earlier structures. Analysis of the cryoEM density was enhanced by docking the crystal structures of both the hexon and penton base capsid proteins. CryoEM density was observed for hexon residues missing from the crystal structure that include hypervariable regions and the epitope of a neutralizing monoclonal antibody. Within the penton base, density was observed for the integrin-binding RGD loop missing from the crystal structure and for the flexible beta ribbon of the variable loop on the side of the penton base. The Ad35 fiber is flexible, consistent with the sequence insert in the third beta-spiral repeat. On the inner capsid surface density is revealed at the base of the hexons and below the penton base. A revised model is presented for protein IX within the virion. Well-defined density was assigned to a conserved domain in the N terminus of protein IX required for incorporation into the virion. For the C-terminal domain of protein IX two alternate conformations are proposed, either binding on the capsid surface or extending away from the capsid. This model is consistent with the tolerance of the C terminus for inserted ligands and its potential use in vector retargeting. This structural study increases our knowledge of Ad capsid assembly, antibody neutralization mechanisms, and may aid further improvements in gene delivery to important human cell types. PMID- 15890368 TI - Multiple functions of a feed-forward-loop gene circuit. AB - The feed-forward-loop (FFL), a network motif in genetic regulatory networks, involves two transcription factors (TFs): one regulates the expression of the second, and both TFs regulate the expression of an effector gene. Analysis of FFL design principles has been initiated, but the functional significance of the FFL is still unclear. In theoretical studies so far, the TFs are assumed to interact with different signals, which is common. However, we have found examples of FFLs in Escherichia coli in which both TFs interact with the same signal. These examples belong to the type 2 incoherent class of FFLs, in which each TF acts exclusively as a repressor of transcription. Here, we analyze mathematical models of this class of circuits, examining a comprehensive array of subclasses that differ in the way a signal modulates the activities of the TFs. Through parameter variation, we characterize statistically how input/output (I/O) behavior and temporal responsiveness are predicted to depend on the wiring of signal interactions in a circuit. We find that circuits can exhibit any of 13 qualitatively distinct steady-state I/O patterns, including inducible and repressible patterns. Some subclasses exhibit as many as six patterns. Transient pulses are also possible, and the response of a circuit to a signal may be either faster or slower than that of a gene circuit in which there is only one TF. Our results provide a catalog of functions for a class of FFL circuits, whose subclasses have different breadths of possible behaviors and different typical behaviors. PMID- 15890369 TI - Schizophrenia, slow wave sleep and visuospatial memory: sleep-dependent consolidation or trait-like correlation? PMID- 15890370 TI - Effect of diabetes on aortic nitric oxide synthesis in spontaneously hypertensive rats; does captopril modulate this effect? AB - Nitric oxide (NO) is a potent regulator in the cardiovascular system; it is generated by the nitric oxide synthase (NOS) family of proteins. NO produced in endothelial cells plays a crucial role in vascular functions. The aim of this study was to clarify the effect of diabetes on aortic NO synthesis in a model of genetic hypertension and determine whether captopril modulates this effect. Diabetes was induced in ten weeks old spontaneously hypertensive rats (SHR) by streptozotocin injection. The rats were allocated into 3 groups: control group 1, non-diabetic SHR; group 2, diabetic SHR; group 3, diabetic SHR group receiving captopril at 80 mg/kg in drinking water for 4 weeks. Mean blood pressure (MBP) was measured once a week by tail-cuff method. Aortic NO metabolities (nitrite/nitrate) and endothelial NOS (NOS-3) were assayed by Griess reaction and by immunoblotting and immunohistochemistry, respectively. There was a significant decrease in nitrite/nitrate (NOx) in aortas of diabetic SHR compared with controls. The decrease of aortic NOx in diabetic SHR was accompanied by a decrease in NOS-3 expression. Captopril treatment reduced MBP without affecting either NOx level or NOS-3 expression in aortas of diabetic SHR. We conclude that STZ-induced diabetes decreased NO in aortas of SHR that may reflect endothelial cell dysfunction; captopril administration decreased MBP without affecting NO level in aortas of diabetic SHR which suggest that the blood pressure-lowering effects of captopril were independent of NO. PMID- 15890371 TI - Evidence for cytochrome P450 2E1 catalytic activity and expression in rat blood lymphocytes. AB - Studies initiated to characterize cytochrome P450 2E1(CYP2E1) in freshly isolated rat blood lymphocytes revealed significant mRNA of CYP2E1 in control blood lymphocytes. RT-PCR studies have shown that as observed in liver, acute treatment of ethanol (single oral dose of 0.8 ml/kg b.wt, i.p), resulted in increase in the mRNA expression of CYP2E1 in freshly isolated rat blood lymphocytes. Western blotting studies using polyclonal antibody raised against rat liver CYP2E1 demonstrated significant immunoreactivity, comigrating with the liver isoenzyme, in freshly isolated control rat blood lymphocytes. Similar to that seen in liver, pretreatment of ethanol was found to produce an increase in the CYP2E1 isoenzyme in the blood lymphocytes. Blood lymphocytes were also found to catalyze the CYP dependent N-demethylation of N-nitrosodimethylamine (NDMA), which like in liver increased 2-3 fold following pretreatment of rats with known CYP2E1 inducers. Kinetic studies have further shown significant increase in the apparent Vmax and the affinity towards the substrate in rat blood lymphocytes indicating that as observed in liver, the increase in mRNA and protein expression following exposure to CYP2E1 inducers is associated with the increased catalytic activity of CYP2E1 in freshly isolated rat blood lymphocytes. The data indicating similarities of the blood lymphocyte CYP2E1 with the liver enzyme suggest that lymphocyte CYP2E1 levels in freshly isolated rat blood lymphocytes could be used to monitor tissue enzyme levels. PMID- 15890372 TI - Mechanisms of mu opioid receptor/G-protein desensitization in brain by chronic heroin administration. AB - Previous studies have shown that chronic opiate treatment decreases mu opioid stimulated [35S]GTPgammaS binding in specific brain regions. To extend these findings, the present study investigated DAMGO-stimulated [35S]GTPgammaS binding in membrane homogenates and coronal sections from rats non-contingently administered heroin. Rats were administered saline or increasing doses of heroin i.v. hourly up to 288 mg/kg/day over 40 days. In brain sections, chronic heroin administration decreased DAMGO-stimulated [35S]GTPgammaS binding in medial thalamus and amygdala, with no effect in cingulate cortex or nucleus accumbens. Chronic heroin administration also reduced [35S]GTPgammaS binding stimulated by the principal metabolite of heroin, 6-monoacetylmorphine. In contrast, no significant changes in mu opioid receptor binding were observed in amygdala or thalamus using [3H]DAMGO autoradiography. In membranes from amygdala and thalamus, chronic heroin treatment decreased the maximal effect of DAMGO in stimulating [35S]GTPgammaS binding, with no effect on DAMGO potency. GTPgammaS saturation analysis showed that chronic heroin treatment decreased the Bmax, and increased the K(D), of DAMGO-stimulated [35S]GTPgammaS binding. These data suggest potential mechanisms by which chronic agonist treatment produces opioid receptor/G-protein desensitization in brain. PMID- 15890373 TI - How do cross-migration models arise? AB - In this paper we present a general method to derive spatio-temporal population models mechanistically. We consider a system of multiple species living in a patchy habitat in which the local population of each species consists of some behavioural groups. We then formulate a continuous-time model where a small positive parameter is present, measuring the time scale of behavioural transitions relative to that of giving birth, death and migration among patches. By the singular perturbation method the model is reduced to a lower dimensional one in which the migration terms are, in general, nonlinear and related to the reaction terms describing the local dynamics. Two examples demonstrating the emergence of cross-migration models, i.e., the models in which the per-capita migration rate of one species depends on the density of some other species, are given. PMID- 15890374 TI - Germline mutations of the MYH gene in Japanese patients with multiple colorectal adenomas. AB - Germline mutations of the MYH gene have been revealed to associate with the recessive inheritance of multiple colorectal adenomas in Caucasian population. However, MYH mutations in Japanese patients have not yet been clarified. In an assessment of MYH mutations, we examined 35 Japanese patients with multiple colorectal adenomas who had neither dominant inheritance of colorectal tumors, nor germline APC mutations. One patient had a homozygous biallelic MYH mutation, R231C and three independent patients had monoallelic MYH mutations at a splice site on exon 11 (IVS10-2 A to G). These four patients had 21 to around 100 colorectal adenomas and 1-3 synchronous colorectal carcinomas. The most common mutations in Caucasian patients, Y165C and G382D, were not detected in our Japanese cases. The MYH mutations detected in Japanese patients were novel and different from those detected among Caucasian, Indian and Pakistani patients, which suggests the existence of ethnic differentiation in MYH mutations. PMID- 15890375 TI - Comparison of the expression profiles induced by genotoxic and nongenotoxic carcinogens in rat liver. AB - Application of recently developed gene expression techniques using microarrays in toxicological studies (toxicogenomics) facilitate the interpretation of a toxic compound's mode of action and may also allow the prediction of selected toxic effects based on gene expression changes. In order to test this hypothesis, we investigated whether carcinogens at doses known to induce liver tumors in the 2 year rat bioassay deregulate characteristic sets of genes in a short term in vivo study and whether these deregulated genes represent defined biological pathways. Male Wistar rats were dosed with the four nongenotoxic hepatocarcinogens methapyrilene (MPy, 60 mg/kg/day), diethylstilbestrol (DES, 10 mg/kg/day), Wy 14643 (Wy, 60 mg/kg/day), and piperonylbutoxide (PBO, 1200 mg/kg/day). After 1, 3, 7, and 14 days, the livers were taken for histopathological evaluation and for analysis of the gene expression profiles on Affymetrix RG_U34A arrays. The expression profile of the four nongenotoxic carcinogens were compared to the profiles of the four genotoxic carcinogens 2-nitrofluorene (2-NF), dimethylnitrosamine (DMN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and aflatoxin B1 (AB1) from a similar study reported previously. By using statistical and clustering tools characteristically deregulated genes were extracted and functionally classified. Distinct cellular pathways were affected by the nongenotoxic carcinogens compared to the genotoxic carcinogens which at least partly correlated with the two-stage model of carcinogenesis. Characteristic to genotoxic carcinogens were a DNA damage response and the activation of proliferative and survival signaling. Nongenotoxic carcinogens showed responses to oxidative DNA or protein damage, as well as cell cycle progression and signs of regeneration. Many of the gene alterations found with the nongenotoxic carcinogens imply compound-specific mechanisms. Although neither a single gene nor a single pathway will be sufficient to discriminate the two classes of carcinogens, it became evident that combinations of pathway-associated gene expression profiles may be used to predict a genotoxic or nongenotoxic carcinogenic potential of a compound in short-term studies. PMID- 15890376 TI - Stepwise mutation likelihood computation by sequential importance sampling in subdivided population models. AB - An importance sampling algorithm for computing the likelihood of a sample of genes at loci under a stepwise mutation model in a subdivided population is developed. This allows maximum likelihood estimation of migration rates between subpopulations. The time to the most recent common ancestor of the sample can also be computed. The technique is illustrated by an analysis of a data set of Australian red fox populations. PMID- 15890377 TI - Inhibition of P-glycoprotein-mediated transport by extracts of and monoterpenoids contained in Zanthoxyli fructus. AB - Citrus (rutaceous) herbs are often used in traditional medicine and Japanese cuisine and can be taken concomitantly with conventional medicine. In this study, the effect of various citrus-herb extracts on P-glycoprotein (P-gp)-mediated transport was examined in vitro to investigate a possible interaction with P-gp substrates. Component monoterpenoids of the essential oil in Zanthoxyli fructus was screened to find novel P-gp inhibitors. LLC-GA5-COL150 cells transfected with human MDR1 cDNA encoding P-gp were used. Cellular accumulation of [3H]digoxin was measured in the presence or absence of P-gp inhibitors or test samples. Aurantii fructus, Evodiae fructus, Aurantii fructus immaturus, Aurantii nobilis pericarpium, Phellodendri cortex, and Zanthoxyli fructus were extracted with hot water (decocted) and then fractionated with ethyl acetate. The cell to medium ratio of [3H]digoxin accumulation increased significantly in the presence of the decoction of Evodiae fructus, Aurantii nobilis pericarpium, and Zanthoxyli fructus, and the ethyl acetate fraction of all citrus herbs used. The ethyl acetate fraction of Zanthoxyli fructus exhibited the strongest inhibition of P-gp among tested samples with an IC50 value of 166 microg/mL. Then its component monoterpenoids, geraniol, geranyl acetate, (R)-(+)-limonene, (R)-(+)-linalool, citronellal, (R)-(+)-citronellal, DL-citronellol, (S)-(-)-beta-citronellol, and cineole, were screened. (R)-(+)-citronellal and (S)-(-)-beta-citronellol inhibited P-gp with IC50 values of 167 microM and 504 microM, respectively. These findings suggest that Zanthoxyli fructus may interact with P-gp substrates and that some monoterpenoids with the relatively lower molecular weight of about 150 such as (R)-(+)-citronellal can be potent inhibitors of P-gp. PMID- 15890378 TI - Taurine protects against methotrexate-induced toxicity and inhibits leukocyte death. AB - The efficacy of methotrexate (MTX), a widely used cytotoxic chemotherapeutic agent, is often limited by severe side effects and toxic sequelae. Regarding the mechanisms of these side effects, several hypotheses have been put forward, among which oxidative stress is noticeable. The present study was undertaken to determine whether taurine, a potent free radical scavenger, could ameliorate MTX induced oxidative injury and modulate immune response. Following a single dose of methotrexate (20 mg/kg), either saline or taurine (50 mg/kg) was administered for 5 days. After decapitation of the rats, trunk blood was obtained and the ileum, liver, and kidney were removed to measure malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity, and collagen content, as well as histological examination. Our results showed that MTX administration increased the MDA, MPO activity, and collagen contents and decreased GSH levels in all tissues (P < 0.001), while these alterations were reversed in taurine-treated group (P < 0.05-0.01). Elevated (P < 0.001) TNF-alpha level observed following MTX treatment was depressed with taurine (P < 0.01). Oxidative burst of neutrophils stimulated by phorbol myristate acetate was reduced in saline-treated MTX group (P < 0.001), while taurine abolished this effect. Similarly, flow cytometric measurements revealed that leukocyte apoptosis and cell death were increased in MTX-treated animals, while taurine reversed these effects (P < 0.05). Reduced cellularity in bone marrow samples of MTX-treated group (P < 0.01) was reversed back to control levels in taurine-treated rats. Severe degeneration of the intestinal mucosa, liver parenchyma, glomerular, and tubular epithelium observed in saline-treated group was improved by taurine treatment. In conclusion, it appears that taurine protects against methotrexate-induced oxidant organ injury and inhibits leukocyte apoptosis and may be of therapeutic potential in alleviating the systemic side effects of chemotherapeutics. PMID- 15890379 TI - A model-based inverse method for positioning scatterers in a cladded component inspected by ultrasonic waves. AB - Nondestructive methods aim at detecting, locating and identifying defects. Inversion of ultrasonic measurements obtained by inspecting a steel component of regular geometry with an immersed transducer leads to accurate location of defects. When the component is cladded, the irregular geometry of the surface and the anisotropic nature of the cladding material lead to aberrations of the radiated field (e.g., beam distortions, splitting and defocusing, these varying with the transducer scanning position). As a consequence, defect location may be inaccurate and defects (e.g., cracks) sizing impossible. In the present paper, a model-based inverse method is developed to solve this problem. It relies on the time-dependent simulation of ultrasonic propagation in this material of complex geometry and structure, in order to determine a set of probable positions for the defect at the origin of the measured ultrasonic echo-structure. The most probable position is determined by minimizing a cost-function of likeness between the simulated and measured ultrasonic images. The overall scheme shall generally apply to inverse measured ultrasonic echo-structures as soon as the simulation of the forward problem is tractable. To validate the method, examples of application are given dealing with actual measurements obtained in the real configuration of pressure vessel inspection. PMID- 15890380 TI - Bubble oscillation and inertial cavitation in viscoelastic fluids. AB - Non-linear acoustic oscillations of gas bubbles immersed in viscoelastic fluids are theoretically studied. The problem is formulated by considering a constitutive equation of differential type with an interpolated time derivative. With the aid of this rheological model, fluid elasticity, shear thinning viscosity and extensional viscosity effects may be taken into account. Bubble radius evolution in time is analyzed and it is found that the amplitude of the bubble oscillations grows drastically as the Deborah number (the ratio between the relaxation time of the fluid and the characteristic time of the flow) increases, so that, even for moderate values of the external pressure amplitude, the behavior may become chaotic. The quantitative influence of the rheological fluid properties on the pressure thresholds for inertial cavitation is investigated. Pressure thresholds values in terms of the Deborah number for systems of interest in ultrasonic biomedical applications, are provided. It is found that these critical pressure amplitudes are clearly reduced as the Deborah number is increased. PMID- 15890381 TI - Grouping local orientation and direction signals to extract spatial contours: empirical tests of "association field" models of contour integration. AB - Over the last decade or so a great deal of psychophysical research has attempted to delineate the principles by which local orientations and motions are combined across space to facilitate the detection of simple spatial contours. This has led to the development of "association field" models of contour detection which suggest that the strength of linking between neighbouring elements in an image, is determined by the degree to which they aligned along smooth (first-order) curves. To test this assumption we used a path detection paradigm to compare the ability of observers to identify the presence of contours defined by either spatial orientation, motion direction or by specific combinations of both types of visual attribute. The relative alignment of the local orientations and/or directions with respect to the axis of the depicted contour was systematically varied. For orientation-defined contours detection was best when the elements were aligned along (parallel with) the contour axis, approached chance levels for obliquely oriented elements and then improved for elements that were orthogonal to the contour axis (i.e., performance was a U-shaped function of degree of orientation misalignment). This pattern of results was found for both straight and curved contours and is not readily explicable in terms of current association field theories. For motion-defined contours, however, performance simply deteriorated as the relative directions of the constituent path elements were progressively misaligned with respect to the contour. Thus the rules by which local orientations are linked to define spatial contours are qualitatively different from those used for linking local directions and each may be mediated by distinct visual mechanisms. When both orientation and motion cues were simultaneously available, contour detection performance was generally enhanced, in a manner that is consistent with probability summation. We suggest that association field models of orientation linking may need to be extended in light of the present findings. PMID- 15890382 TI - The adaptive effect of narrowing the interocular separation on the AC/A ratio. AB - The purpose of this study was to determine whether the response AC/A ratio could be altered when the subject's interpupillary distance (IPD) was optically halved. We measured the changes in the AC/A ratio for 10 subjects after using the optical device for 30 min. Accommodative response was measured using a Canon R-1 optometer, and vergence response was measured with an ASL 210 Eye Movement Monitor. The average AC/A ratios were 1.20+/-0.35 (SD) (MA/D) and 0.84+/-0.39 (MA/D) before and after wearing the device, respectively. The decrease in AC/A ratio was statistically significant (p=0.01). This was mainly caused by a reduction in the slope of the accommodative vergence. The results of this study suggest that the AC/A ratio can be decreased if an IPD-narrowing device is used. A possible application of this mechanism in the study of myopia is discussed. PMID- 15890383 TI - Small-scale, hydrogen-oxidizing-denitrifying bioreactor for treatment of nitrate contaminated drinking water. AB - Nitrate removal by hydrogen-coupled denitrification was examined using flow through, packed-bed bioreactors to develop a small-scale, cost effective system for treating nitrate-contaminated drinking-water supplies. Nitrate removal was accomplished using a Rhodocyclus sp., strain HOD 5, isolated from a sole-source drinking-water aquifer. The autotrophic capacity of the purple non-sulfur photosynthetic bacterium made it particularly adept for this purpose. Initial tests used a commercial bioreactor filled with glass beads and countercurrent, non-sterile flow of an autotrophic, air-saturated, growth medium and hydrogen gas. Complete removal of 2 mM nitrate was achieved for more than 300 days of operation at a 2-h retention time. A low-cost hydrogen generator/bioreactor system was then constructed from readily available materials as a water treatment approach using the Rhodocyclus strain. After initial tests with the growth medium, the constructed system was tested using nitrate-amended drinking water obtained from fractured granite and sandstone aquifers, with moderate and low TDS loads, respectively. Incomplete nitrate removal was evident in both water types, with high-nitrite concentrations in the bioreactor output, due to a pH increase, which inhibited nitrite reduction. This was rectified by including carbon dioxide in the hydrogen stream. Additionally, complete nitrate removal was accomplished with wastewater-impacted surface water, with a concurrent decrease in dissolved organic carbon. The results of this study using three chemically distinct water supplies demonstrate that hydrogen-coupled denitrification can serve as the basis for small-scale remediation and that pilot-scale testing might be the next logical step. PMID- 15890384 TI - Concentrations of pesticide residues in grasses and sedges due to point source contamination and the indications for public health risks, Vikuge, Tanzania. AB - The concentrations of organochlorine pesticide residues were determined in roots and leaves of giant sedges (Cyperus exaltatus), fresh leaves and dry hay of guinea grasses (Panicum maximum), and in stems and leaves of elephant grasses (Pennisetum purpureum) in the vicinity of a point source of contamination, an old storage site at Vikuge farm in Tanzania. The GPC-cleaned extracts were analyzed by GC-ECD and GC/MS. The concentrations of total DDT ranged from 83 to 18274 ng/g, 166 to 7922 ng/g, and 68 to 405 ng/g, on fresh weight basis, in C. exaltatus, P. maximum and P. purpureum, respectively. The concentrations of total HCH were up to 74 ng/gfw in leaves of C. exaltatus, 43 ng/gdw in dry hay of P. maximum, and 10 ng/gfw in stems of P. purpureum. Aldrin and dieldrin were only detected in C. exaltatus and their concentrations were up to 11 and 8 ng/gfw, respectively. The strong positive correlations between the concentrations of the detected compounds suggest a common source. The concentrations of total DDT were far above the Australian extraneous maximum residue limit in primary animal feedstuffs. These findings indicate potential risks and concerns for livestock and public health. The concentrations of aldrin, dieldrin and lindane were lower than the Australian limits. PMID- 15890386 TI - Biomonitoring of trace elements in the leaves and fruits of wild olive and holm oak trees. AB - Biomonitoring of trace elements is essential to assess ecosystem health, in particular in landscapes influenced by human activity. The Guadiamar Valley (SW Spain) was polluted in 1998 by a spill from an open-pit pyrite mine affecting about 55 km2. In this paper, we used two common species of tree, namely wild olive and holm oak, to biomonitor the concentration of nine trace elements-As, Cd, Cu, Fe, Mn, Ni, Pb, Tl and Zn-in this spill-affected area over the 3-year period 1999-2001. We analysed the leaves and fruits of trees growing in the spill affected terraces, and compared them with adjacent trees in the non-affected upper terraces. The main trace elements polluting the soil were Zn, As, Pb and Cu. In general, the oak leaves were richer in trace elements than the olive leaves, reaching phytotoxic levels for As and Pb, while the olive fruits (pulp) were more polluted than the oak seeds (protected inside a hard pericarp), reaching toxic values for Cd and Pb. The concentration of trace elements in the leaves and fruits decreased with time and, in consequence, the toxicity risk to the food web diminished. PMID- 15890387 TI - Public health evaluation of cadmium concentrations in liver and kidney of moose (Alces alces) from four areas of Alaska. AB - Liver and/or kidney samples were collected from 139 hunter-killed moose from four areas of Alaska during 1986. The concentration of cadmium in organ tissue was determined by direct-current plasma atomic emission spectrometry. All results are reported as mug/g wet weight. Concentrations of cadmium in liver ranged from 0.06 microg/g to 9.0 microg/g; in the kidney cortex they ranged from 0.10 microg/g to 65.7 microg/g. Cadmium levels were significantly associated with location and age. The highest geometric mean liver (2.11 microg/g) and kidney cortex (20.2 microg/g) cadmium concentrations were detected in moose harvested near Galena, Alaska. Limited dietary information from Alaska and Canada indicates that the intake of moose liver or kidney does not exceed, in most individuals, the World Health Organization recommendations for weekly cadmium consumption of 400 microg to 500 microg. Additionally, human biomonitoring data from Canada and Alaska indicate exposure to cadmium is low except for individuals who smoke cigarettes. Given the nutritional and cultural value of subsistence foods, the Alaska Division of Public Health continues to support the consumption of moose liver and kidney as part of a well-balanced diet. Human biomonitoring studies are needed in Alaska to determine actual cadmium exposure in populations with a lifelong history of moose liver and kidney consumption. PMID- 15890388 TI - Geochemical background--concept and reality. AB - The definitions and use of the term 'background' in exploration and environmental geochemistry are reviewed. Based on data from two subcontinental-scale geochemical mapping projects, it is shown that trying to define 'a background' for a large area is fraught with problems. It is demonstrated that background may change from area to area within a region and between regions. Although global averages are of general use, no specific global background levels of elements, for example in soils, can be defined, at best regional or local operational estimates can be made, though with caveats. Using background estimates based on concentrations in deeper soil levels to judge element concentrations in upper soil horizons (e.g., the TOP/BOT-ratio) can lead to severe misinterpretations if natural biogeochemical soil formation processes are ignored. Because of large natural variations in element concentrations in, for example soils, even the establishment of maximum admissible concentration based on ecotoxicological investigations is a difficult exercise. Organisms may become adapted to natural differences. Furthermore, there are challenges in converting the concentrations of the soluble substances used in ecotoxicological studies to appropriate levels in solid phase material, for example soils, analysed by commonly employed acid digestion procedures. Toxicological thresholds may thus also need to consider a spatial component that is presently neglected. PMID- 15890389 TI - WITHDRAWN: NIH conference on dietary supplements. "American Society of Health System Pharmacists" AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 15890390 TI - Relationship between soluble thrombomodulin in patients with intermittent claudication and critical ischemia. AB - INTRODUCTION: Thrombomodulin (TM) has been described as a marker of endothelial injury in atherosclerosis. The role of TM as a predictor of PAD severity is to be proven. The goal of the present study is to compare the level of plasmatic (TMp) in patients with intermittent claudication with patients with critical ischemia in the lower limbs. MATERIALS AND METHODS: TMp was measured using ELISA in the plasma of 41 patients with intermittent claudication degree 1 and in 40 patients presenting critical ischemia in the lower limbs degrees 2 and 3, according to TASC. The hypotheses of normality and homogeneity of the variance had been proven via Shapiro-Wilk and Levene tests, respectively. The comparison of the TMp between the groups was done using the t-Student test. RESULTS: No statistically significant difference was observed. The average levels of TMp for intermittent claudication were 5.2 ng/ml (0.78-13.61 ng/ml) and TMp for critical ischemia in the lower limbs were 6.34 (0.82-18.22 ng/ml) where p=0.265. CONCLUSION: TMp does not seem to be an appropriate marker for PAD severity. PMID- 15890391 TI - Comparison of levels of inflammatory markers and hemostatic factors in the patients with and without peripheral arterial disease. AB - INTRODUCTION: Peripheral arterial disease is one aspect of atherosclerosis, a disease associated with both inflammation and hypercoagulability. Many recent studies have focused on the diversity of mechanisms by which inflammation can promote blood clotting. However, the relationship between plasma concentrations of inflammatory and hemostatic markers and the severity of atherosclerosis is not yet well studied. We want to determine the relations among selected D-dimer, C reactive protein, fibrinogen, prothrombin time and serum amyloid A, and the ankle brachial index in patients with and without peripheral arterial disease. MATERIALS AND METHODS: In a prospective cohort study, 45 consecutive patients with peripheral arterial disease of ankle brachial index<0.90, and 44 patients without peripheral arterial disease of ankle brachial index 0.90 to 1.50 were included. D-dimer, fibrinogen, C-reactive protein, serum amyloid A, and prothrombin time were measured at the recruitment. RESULTS: Median values of serum amyloid A, D-dimer, and C-reactive protein were significantly higher in the peripheral arterial disease group than in those without peripheral arterial disease group (p<0.001). The patients with PAD had moderately higher fibrinogen levels than without PAD (p<0.01). Prothrombin time levels were normal in both groups. In multivariable regression analyses adjusting for all blood factors as well as potential confounders, patients with peripheral arterial disease, levels of serum amyloid A, and C-reactive protein showed a highly significant, inverse association with the ankle brachial index. D-dimer and fibrinogen level increase were also found to be related to lower ankle brachial index, while no association was observed between prothrombin time levels. CONCLUSIONS: Higher C-reactive protein, serum amyloid A, and D-dimer levels are showing positive association with the presence of peripheral arterial disease. C-reactive protein and serum amyloid A levels are direct relations between the ankle brachial index and the extent of vascular inflammation. PMID- 15890392 TI - Simple technique for improving tubeless cutaneous ureterostomy. AB - INTRODUCTION: Cutaneous ureterostomy is the simplest and safest method of all permanent urinary diversions. However, the procedure does carry a risk of stomal stenosis. We describe a simple technique for improving tubeless cutaneous ureterostomy. TECHNICAL CONSIDERATIONS: Between June 1991 and June 2003, the Toyoda cutaneous ureterostomy was performed in 54 patients (102 renal units) with a minimum of 6 months of follow-up. Since 1998, we introduced a new technique, consisting of fixation between the anterior and posterior rectus sheath by four interrupted sutures to maintain the stability of the abdominal wall tunnel for the ureters. The ureteral patency rate was reviewed. Of the 79 renal units (77.5%) that achieved a tubeless condition, 70 (68.6%) had no hydronephrosis. The catheter-free rate improved from 60.5% (26 of 43 renal units) to 89.8% (53 of 59 renal units) with the introduction of the new surgical stabilization step for the abdominal wall tunnel. CONCLUSIONS: This surgical modification is an effective and simple procedure that improves tubeless cutaneous ureterostomy performed with the Toyoda method. PMID- 15890393 TI - Percutaneous extraction of bilateral renal mycetomas in premature infant using mechanical thrombectomy device. AB - Although mycotic infections of the urinary system occur not infrequently, obstruction of the upper urinary tract because of mycetomas or fungus balls is exceedingly rare. Treatment consists of antifungal treatment and appropriate urinary drainage. In cases resistant to treatment with external drainage and chemotherapy, open surgery is typically performed to remove the mycetoma. However, this can also be achieved percutaneously using mechanical thrombectomy devices, as shown in this case of a 4-month-old infant presenting with renal failure due to bilateral obstructing mycetomas. PMID- 15890394 TI - Epithelioid leiomyosarcoma of the uterine cervix: a case report and review of the literature. AB - BACKGROUND: Epithelioid leiomyosarcomas arising from the uterine cervix are extremely rare neoplasms, with only three cases reported in the English literature. CASE: A 53-year-old Japanese woman was admitted to our university hospital due to massive vaginal bleeding from a cervical tumor. A total hysterectomy with bilateral salpingo-oophorectomy was performed. Histological findings, including immunohistochemical study using desmin, SMA, cytokeratin, S 100, HMB-45, vimentin, melan-A, and CD68, led to a diagnosis of epithelioid leiomyosarcoma of the uterine cervix. The patient underwent adjuvant chemotherapy and has been disease-free for over 20 months. CONCLUSION: Immunohistochemical studies may be needed to differentiate among the alternative diagnoses of malignant melanoma, metastatic carcinoma, and epithelioid sarcoma. Additional cases are needed to develop optimal management strategies and to predict prognosis. PMID- 15890395 TI - Peritoneal tuberculosis mimicking ovarian cancer can be diagnosed by polymerase chain reaction: a case report. AB - BACKGROUND: Confusion can result when a differential diagnosis includes peritoneal tuberculosis and ovarian cancer because of overlapping clinical and laboratory findings. CASE: A 37-year-old woman presented with aggressive abdominal distension. At laparotomy, massive ascites was present accompanied by miliary deposits throughout the abdomen. Polymerase chain reaction (PCR) conducted with ascites fluid produced DNA sequences compatible with tuberculosis. CONCLUSION: PCR can be a rapid and reliable method for identification of peritoneal tuberculosis; acceleration of the diagnostic decision-making process prevents exposure to unnecessary surgery and allows early initiation of anti tuberculosis treatment. PMID- 15890396 TI - Understanding locally advanced breast cancer: what influences a woman's decision to delay treatment? AB - BACKGROUND: Despite early detection having been identified as a key factor in long-term survival of patients with breast cancer, women with locally advanced breast cancer continued to present at many medical centers. METHODS: Women with locally advanced breast cancer (LABC) (N = 11) and matched controls (N = 11) with Stage I or II breast cancer completed semi-structured interviews and the Hospital Anxiety and Depression Scale, Life Orientation Test, Multidimensional Health Locus of Control Scale, Body Investment Scale, Monitor-Blunter Style Scale, and Religious Coping Scale. Five spouses of the LABC group and eight control partners were similarly assessed. RESULTS: On psychological tests, no significant differences were found between patient groups; however, LABC spouses reported significantly greater active religious surrender (t = 2.37, P = 0.037) and depression (t = 3.54, P = 0.047) than control spouses. The women's semi structured interviews identified that LABC was associated with inattention to routine breast cancer screening, denial, fatalism and reliance on alternative therapies. Spouses of the LABC group tended to be more passive in their wives' medical care, and also utilized fatalistic thinking and denial. CONCLUSIONS: It is important for health care providers to identify patients who will require extra encouragement to seek timely medical care and to follow through with treatment recommendations. PMID- 15890397 TI - Cancer death epidemics in United States Black males: evaluating courses, causation, and cures. AB - BACKGROUND: Estimates that smoking contributes 38-72% of the United States (US) Black male cancer death rate leave a wide range of uncertainty. This paper uses additional and regional data, and refined methods, to reassess that range. METHODS: This study uses lung cancer rates as an exposure index, linear regression, age adjusted US 1950-2001 and US regional 1969-2001 death rates (rates), and the formula: smoking-attributable fraction (SAF)=(1-((rate in the unexposed) / (rate in the exposed))). Estimated lung cancer rates in the unexposed range between rates predicted for a population with no smoking attributable lung cancers to rates seen in "nonsmokers." RESULTS: Lung cancer death rates predicted 99.9% and 99.8% of the variances in non-lung non-stomach cancer death rates from 1950-1980 and 1950-1988, respectively (each P<0.0001). That suggests 2001 all-sites cancer death SAFs of 63% (sensitivity range 60-66%) nationally and from 43% in the Northeast to 67% in the South. CONCLUSIONS: Smoking may cause most premature cancer deaths and temporal and regional cancer death rate disparities in Black men. PMID- 15890398 TI - Successful use of oviduct epithelial cell coculture for in vitro production of viable red deer (Cervus elaphus) embryos. AB - Techniques for in vitro production (IVP) of viable embryos have been thoroughly developed in several domestic species in view to improve breeding efficiency. When applied to wild life, these techniques may also help the maintenance of biodiversity through amplification of sparse animals offspring and facilitation of genetic material exchange. During the successive steps of IVP, i.e. oocyte in vitro maturation (IVM), fertilization (IVF) and early embryo development (IVD) to the blastocyst stage, gametes and embryos are faced with unusual environment, including oxidative stress, known to be detrimental to their survival. In the present study, starting from methods developed in domestic species, we have adapted IVP to produce viable red deer embryos. In a first experiment, cumulus cells were removed from in vitro matured oocytes either before or after IVF. The presence of cumulus cells during IVF did not affect final cleavage or development rates. In a second experiment, in vitro matured oocytes were fertilized in the presence of cumulus cells and cultured in SOFaaBSA medium alone or in the presence of ovine oviduct epithelial cell (oOEC) monolayer. Whereas, oviduct cells did not improve the cleavage rate, they significantly increased the rate of embryos reaching the blastocyst stage (from 3 to 25% of total oocytes). Ten blastocysts from oOEC coculture were transferred after freezing and thawing to five recipient hinds and gave rise to three pregnancies. The three pregnant hinds gave birth to three live and normal calves. PMID- 15890399 TI - Biological fabrication of nacreous coating on titanium dental implant. AB - Titanium screws with 3.5mm diameter and 8mm length, as well as titanium flat sheets with size 4 mm x 8 mm x 0.3mm, were implanted into the epithelial mantle pearl sacs of a fresh water bivalve (Hyriopsis cumingii Lea) by replacing the pearls. After 45 days of cultivation, the implant surfaces were deposited with a nacre coating with iridescent luster. The coating could conform to some extent the thread topography of the screw implant and was about 200-600 microm in thickness. The coating was composed of a laminated nacreous layer and a transitional non-laminated layer that consisted mainly of vaterite and calcite polymorphs of calcium carbonate. The transitional layer was around 2-10 microm thick in the convex and flat region of the implant surface and could form close contact with titanium surface; while the transitional layer was much thicker in the steep concave regions and could not form close contact with the titanium surface. The reasons for inhomogeneity in thickness and the variation in interface character were discussed and the improvement to the design of the dental implant with respect to this coating method was suggested in the paper. The results suggest that it is possible to fabricate a biologically active and degradable, and mechanically tough and strong nacre coating on titanium dental implant by this novel coating technology. PMID- 15890400 TI - Protein binding properties of surface-modified porous polyethylene membranes. AB - In this study, we quantified the adsorption of immunoglobulin G (IgG) protein onto several polyelectrolyte-modified sintered porous polyethylene (PPE) membranes. The polymer surfaces had both cationic and anionic charges obtained via the adsorption of polyethylenimine (PEI) and polyacrylic acid (PAA), respectively, onto plasma-activated PPE. The amount of IgG adsorption was determined by measuring the gamma radiation emitted by [125I]-IgG radio labeled protein. By studying the impact of pH and ionic strength on IgG adsorption, we attempted to characterize the role and nature of the electrostatic interactions involved in the adsorption process to better understand how these interactions were influenced by the charge and structure of immobilized polyelectrolyte complexes at modified membrane surfaces. We were able to show that surface modification of PPE membranes with adsorbed PEI monolayers and PEI-PAA bilayers can greatly improve the IgG binding ability of the membrane under optimized conditions. We also showed that the observed improvement in the IgG binding is derived from electrostatic interactions between IgG and the polyelectrolyte surface. In addition, we found that the greatest IgG adsorption occurred when the IgG and the surface possessed predominantly opposite charges, rather than when the surface possessed the greatest electrostatic charge. Finally, we have found that the molecular weight of the terminating polyelectrolyte has a noticeable effect upon the electrostatic interactions between IgG and the PEI-PAA bilayer modified PPE surfaces. PMID- 15890401 TI - Slow crack growth behaviour of hydroxyapatite ceramics. AB - Among materials for medical applications, hydroxyapatite is one of the best candidates in orthopedics, since it exhibits a composition similar to the mineral part of bone. Double torsion technique was here performed to investigate slow crack growth behaviour of dense hydroxyapatite materials. Crack rate, V, versus stress intensity factor, K(I), laws were obtained for different environments and processing conditions. Stress assisted corrosion by water molecules in oxide ceramics is generally responsible for slow crack growth. The different propagation stages obtained here could be analyzed in relation to this process. The presence of a threshold defining a safety range of use was also observed. Hydroxyapatite ceramics appear to be very sensitive to slow crack growth, crack propagation occurring even at very low K(I). This can be explained by the fact that they contain hydroxyl groups (HAP: Ca(10)(PO(4))(6)(OH)(2)), favouring water adsorption on the crack surface and thus a strong decrease of surface energy in the presence of water. This study demonstrates that processing conditions must be carefully controlled, specially sintering temperature, which plays a key role on V-K(I) laws. Sintering at 50 degrees C above or below the optimal temperature, for example, may shift the V-K(I) law towards very low stress intensity factors. The influence of ageing is finally discussed. PMID- 15890402 TI - Protective effect of colostrinin on neuroblastoma cell survival is due to reduced aggregation of beta-amyloid. AB - Colostrinin (CLN), a mixture of proline-rich polypeptides, has shown a stabilizing effect on cognitive function in Alzheimer's patients measured by the Alzheimer's disease Assessment Scale-cognitive (ADAS-cog) and in Instrumental Activities of Daily Living (ILDL) in recently conducted clinical trials. The aim of this study was to elucidate a possible mode of action of CLN in the treatment of Alzheimer's disease. Here, we report that CLN prevents the aggregation of beta amyloid peptide Abeta (1-40) in vitro. The impact of CLN on the fibril formation was monitored by optical and electron microscopy. The electron micrographs illustrate that, at 25 microM, Abeta (1-40) peptides formed fibrils after 24-48 h of incubation. The presence of 0.25 microM CLN completely abolished the fibril formation. Abeta (1-40) peptides grow into dense fibers when examined at the 20th day. In the presence of CLN, however, the fibrils are much shorter and less dense. Addition of CLN as late as the 17th day can still dissolves the preformed fibrils. These observations were compared to the effect of CLN on the neurotoxic activity of beta-amyloid peptides in the cell culture model (SHSY-5Y). The beta amyloid peptides were pre-incubated with CLN at various times and used to treat SHSY-5Y neuroblastoma cells for up to 4 days. The cytotoxic effect was monitored by trypan blue exclusion. We demonstrated that 24-48 h treatment was the onset of toxicity of 10-50 microM of beta-amyloid peptides. Pre-incubation of 0.0025 0.25muM of CLN with 25 microM of beta-amyloid peptides leads to near-complete abolition of cytotoxicity. Low doses of CLN (2.5 nM) can attain cytotoxic protection levels similar to those of highest doses (0.25 microM). Thus, the time course for the appearance of beta-amyloid fibrils coincides with that for cytotoxicity, and that the reduction of fibrils of beta-amyloid peptides by CLN is concomitant with the reduction of the cytotoxic effects of beta-amyloid on SHSY-5Y neuroblastoma cells. Our studies suggest that the neuroprotective effects exerted by CLN are related to the reduction of beta-amyloid fibrils. PMID- 15890403 TI - Recent advances in animal models of chronic antidepressant effects: the novelty induced hypophagia test. AB - Animal models exhibiting sensitivity to chronic, but not acute, antidepressant treatment are greatly needed for studying the neural mechanisms of the antidepressant response. Although several models of acute antidepressant effects provide excellent tools for antidepressant discovery, they do not permit investigation into their therapeutic effects, which require several weeks of treatment to emerge. The inhibition of feeding produced by novelty, termed 'hyponeophagia', provides an anxiety-related measure that is sensitive to the effects of chronic, but not acute or subchronic, antidepressant treatment. This review evaluates the value of hyponeophagia-based tests as tools for investigating the neurobiology of the therapeutic response to antidepressant treatment. Criteria for the development and validation of animal models used to study neurobiological mechanisms of the antidepressant response are presented. Methodological considerations affecting the reliability, specificity, and ease of use of hyponeophagia-based models are also discussed. Lastly, we present a newly revised hyponeophagia paradigm, called the novelty-induced hypophagia (NIH) test, which attempts to maximize the predictive validity and practicality of the test. The NIH paradigm provides a promising new model for investigations into the neurobiology underlying the antidepressant response. PMID- 15890404 TI - The tail suspension test as a model for assessing antidepressant activity: review of pharmacological and genetic studies in mice. AB - Since its introduction almost 20 years ago, the tail suspension test has become one of the most widely used models for assessing antidepressant-like activity in mice. The test is based on the fact that animals subjected to the short-term, inescapable stress of being suspended by their tail, will develop an immobile posture. Various antidepressant medications reverse the immobility and promote the occurrence of escape-related behaviour. This review focuses on the utility this test as part of a research program aimed at understanding the mechanism of action of antidepressants. We discuss the inherent difficulties in modeling depression in rodents. We describe how the tail suspension differs from the closely related forced swim test. Further, we address some key issues associated with using the TST as a model of antidepressant action. We discuss issues regarding whether it satisfies criteria to be a valid model for assessing depression-related behavioural traits. We elaborate on the tests' ease of use, strain differences observed in the test and gender effects in the test. We focus on the utility of the test for genetic analysis. Furthermore, we discuss the concept of whether immobility maybe a behavioural trait relevant to depression. All of the available pharmacological data using the test in genetically modified mice is collated. Special attention is given to selective breeding programs such as the Rouen 'depressed' mice which have been bred for high and low immobility in the tail suspension test. We provide an extensive pooling of the pharmacological studies published to date using the test. Finally, we provide novel pharmacological validation of an automated system (Bioseb) for assessing immobility. Taken together, we conclude that the tail suspension test is a useful test for assessing the behavioural effects of antidepressant compounds and other pharmacological and genetic manipulations relevant to depression. PMID- 15890405 TI - Complement before molecular biology. AB - Complement activity was first described in the 1890s and the characterisation of this highly complex system has continued ever since. This review traces the history of complement research from its beginnings until it was transformed by the advent of molecular biology in the 1980s. It takes as a focus point the CIBA symposium on Complement held in London in May 1964 and reflects-and is slanted by the views and research experience of the author. PMID- 15890406 TI - Acute effects of 6-hydroxydopamine on dopaminergic neurons of the rat substantia nigra pars compacta in vitro. AB - 6-Hydroxydopamine (6-OHDA) is a neurotoxin which has been implicated in the degeneration of dopaminergic neurons of the substantia nigra pars compacta (SNc) in Parkinson's disease (PD), and is frequently used to produce animal models of the disease. The aim of our study, conducted on midbrain slices obtained from young Wistar rats, was to determine the little known acute effects of this toxin (0.2-2.0 mM; 10-20 min exposure; 34 degrees C) on electrophysiological properties, intracellular Ca2+ levels and dendritic morphology of SNc neurons. Four experimental approaches were used: extracellular recording of firing frequency, whole-cell patch-clamping, ratiometric fura-2 imaging, and cell labeling with lucifer yellow (LY) or dextran-rhodamine. Extracellular recording revealed a concentration-dependent decrease in the tonic, pacemaker-like firing. In whole-cell recordings in voltage-clamp (V(hold) -60 mV), smaller doses (0.2 0.5 mM) induced an outward current (or cell membrane hyperpolarization in current clamp), which could in some cells be reversed with tolbutamide (blocker of ATP dependent K+ channels). A higher dose (1.0-2.0 mM) caused rapid reductions of cell membrane capacitance and membrane resistance. Toxin exposure gradually increased the intracellular Ca2+ level, which did not subsequently return to control. The increase in Ca2+ signal was not prevented by depletion of intracellular Ca2+ stores with thapsigargin (10 microM) or cyclopiazonic acid (30 microM), nor by removing extracellular Ca2+. Cell membrane current and Ca2+ responses were not prevented by blocking dopamine transporter (DAT). Cells loaded with LY or dextran-rhodamine showed signs of damage (cell membrane blebbing) in dendrites following toxin exposure (1 mM; 10-20 min). These results demonstrate that the oxidative and metabolic stress induced in SNc neurons by 6-OHDA results in rapid dose-dependent changes of cell membrane properties with morphological evidence of dendritic damage, as well as in disturbance of intracellular Ca2+ homeostasis. PMID- 15890407 TI - Comparison of neuroprotective effects induced by alpha-phenyl-N-tert-butyl nitrone (PBN) and N-tert-butyl-alpha-(2 sulfophenyl) nitrone (S-PBN) in lithium pilocarpine status epilepticus. AB - The status epilepticus (SE) induced in rats by lithium-pilocarpine (Li-pilo) shares many common features with soman-induced SE including extensive limbic neuropathology. Reactive oxygen species are hypothesized to play a role in the SE induced neuropathology and we propose that the free radical scavengers alpha phenyl-N-tert-butyl nitrone (PBN) and N-tert-butyl-alpha-(2 sulfophenyl) nitrone (S-PBN) may be neuroprotective. PBN or S-PBN were administered either immediately following pilocarpine (exposure treatment) or 5 min after the onset of SE as determined by ECoG activity. SE was allowed to continue for 3 h before termination with propofol. The rats were sacrified 24 h following pilocarpine administration. S-PBN induced minor effects to reduce SE duration and improve neurological deficit 24 h following pilocarpine administration. One hundred and fifty milligrams per kilograms PBN administered 5 min after SE onset produced significant neuroprotection in the parietal, occipital, perirhinal and piriform cortices as well as the lateral amygdala. One hundred and fifty milligrams per kilograms S-PBN was neuroprotective only in the occipital and perirhinal cortex while 300 mg/kg S-PBN exacerbated cortical neuropathology. S-PBN administered 5 min after SE onset exacerbated neuropathology in thalamic regions. In contrast, PBN and S-PBN administered as exposure treatment exacerbated neuropathology in thalamic and CA3 regions. The differential neuroprotective effects of PBN and S PBN may be the result of the poor brain penetration by S-PBN. The results suggest that free radical scavenger activity is neuroprotective in cortical regions during cholinergic convulsions. Regional variations in drug-induced neuroprotectant activity in Li-pilo SE are common and suggest multiple mechanisms of neuropathology. PMID- 15890408 TI - Pharmacogenetics for individualized cancer chemotherapy. AB - The same doses of medication cause considerable heterogeneity in efficacy and toxicity across human populations. Genetic factors are thought to represent important determinants of drug efficacy and toxicity. Pharmacogenetics focuses on the prediction of the response of tumor and normal tissue to standard therapy by genetic profiling and, thereby, to select the most appropriate medication at optimal doses for each individual patient. In the present review, we discuss the relevance of single nucleotide polymorphisms (SNP) in genes, whose gene products act upstream of the actual drug target sites, that is, drug transporters and drug metabolizing phase I and II enzymes, or downstream of them, that is, apoptosis regulating genes and chemokines. SNPs in relevant genes, which encode for proteins that interact with anticancer drugs, were also considered, that is, enzymes of DNA biosynthesis and metabolism, DNA repair enzymes, and proteins of the mitotic spindle. A significant body of evidence supports the concept of predicting drug efficacy and toxicity by SNP genotyping. As the efficacy of cancer chemotherapy, as well as the drug-related toxicity in normal tissues is multifactorial in nature, sophisticated approaches such as genome-wide linkage analyses and integrate drug pathway profiling may improve the predictive power compared with genotyping of single genes. The implementation of pharmacogenetics into clinical routine diagnostics including genotype-based recommendations for treatment decisions and risk assessment for practitioners represents a challenge for the future. PMID- 15890409 TI - HIV-dementia, Tat-induced oxidative stress, and antioxidant therapeutic considerations. AB - Oxidative stress is thought to play a role in the onset of dementia. HIV-dementia has recently been demonstrated to be associated with oxidative stress as indexed by increased protein and lipid peroxidation in the brain and cerebrospinal fluid compared to HIV non-demented patients. The HIV protein Tat induces neurotoxicity, and, more recently, Tat was found to induce oxidative stress directly and indirectly. The role of Tat in HIV-dementia and possible therapeutic strategies involving endogenous and exogenous antioxidants are discussed. PMID- 15890410 TI - Brain hyperthermia as physiological and pathological phenomena. AB - Although brain metabolism consumes high amounts of energy and is accompanied by intense heat production, brain temperature is usually considered a stable, tightly "regulated" homeostatic parameter. Current research, however, revealed relatively large and rapid brain temperature fluctuations (3-4 degrees C) in animals during various normal, physiological, and behavioral activities at stable ambient temperatures. This review discusses these data and demonstrates that physiological brain hyperthermia has an intra-brain origin, resulting from enhanced neural metabolism and increased intra-brain heat production. Therefore, brain temperature is an important physiological parameter that both reflects alterations in metabolic neural activity and affects various neural functions. This work also shows that brain hyperthermia may be induced by various drugs of abuse that cause metabolic brain activation and impair heat dissipation. While individual drugs (i.e., heroin, cocaine, methamphetamine, MDMA) have specific, dose-dependent effects on brain and body temperatures, these effects are strongly modulated by an individual's activity state and environmental conditions, and change dramatically during the development of drug self-administration. Thus, brain thermorecording may provide new information on the central effects of various addictive drugs, drug-activity-environment interactions in mediating drugs' adverse effects, and alterations in metabolic neural activity associated with the development of drug-seeking and drug-taking behavior. While ambient temperatures and impairment of heat dissipation may also affect brain temperature, these environmental conditions strongly potentiate thermal effects of psychomotor stimulant drugs, resulting in pathological brain overheating. Since hyperthermia exacerbates drug-induced toxicity and is destructive to neural cells and brain functions, use of these drugs under activated conditions that restrict heat loss may pose a significant health risk, resulting in both acute life-threatening complications and chronic destructive CNS changes. PMID- 15890411 TI - Decomposing rhythmic hippocampal data to obtain neuronal correlates. AB - Characterizing hippocampal electrical rhythmic activities requires a broadly applicable methodology that lends itself to physiological interpretation. In the intact hippocampal preparation, spontaneous rhythmic field potentials are exhibited in the 3--4 Hz range which evidence suggests is due to discharges in the inhibitory interneuron population. Because field rhythms arise as a network effect and models must be built from the neuron up, we focus on developing a methodology to de-construct the non-stationary rhythms into its important constituents. This study uses 50 CA1/CA3 local field potentials to determine the important constituents, and an additional field recording and two intracellular recordings are examined subsequently. We determine the suitability of several time-frequency techniques. Distinct regions in the time-frequency domain which account for the signal behaviour are then characterized in terms of duration and frequency. These characteristics are interpreted as arising from a statistical mixture distribution. The decomposition of the 50 recordings yields three components whose patterns of activity match those of the intracellular recordings. We suggest that the statistical variability of the local field data can be linked to the variability of neuronal activities seen in intracellular data. PMID- 15890412 TI - Self-reported psychotic symptoms predict impulsivity among African-American patients in an urban non-psychiatric medical setting. AB - A variety of epidemiological studies have documented self-reported psychotic symptoms among individuals in the general population. Research has not been conducted on the associations between self-reported psychotic symptoms and enduring personality characteristics, such as impulsivity, among participants in non-psychiatric settings. We hypothesized that impulsivity scores, as measured by the Barratt Impulsiveness Scale (BIS-11), would be predicted partly by the presence of one or more of a variety of positive psychotic experiences, determined by the revised Symptom Checklist-90. The sample consisted of 100 African-American men and women seeking care in medical clinics in an urban public sector hospital. The sample was divided into those participants endorsing one or more of six psychotic symptoms and those not reporting psychotic symptoms. Multiple linear regression models examined predictors of impulsivity (as measured by the BIS-11 total score and the two derived subscales), including the effect of self-reported positive psychotic symptoms. The presence of psychotic symptoms was predictive of the total impulsivity score and the ideo-motor impulsivity subscale score, even after adjustment for the effects of other correlates of impulsivity, including gender, homelessness, history of conviction for a misdemeanor or felony, and history of past treatment or hospitalization for psychiatric or substance abuse problems. The findings suggest that there is an important link between the presence of self-reported psychotic symptoms and impulsivity, especially ideo-motor impulsivity, in a general sample of low-income African American men and women seeking ambulatory medical care. Further research on self reported psychotic symptoms in non-psychiatric populations is needed, as well as research on the personality correlates of such symptoms, including impulsivity. PMID- 15890413 TI - Condensed version of the Quality of Life Scale for schizophrenia for use in outcome studies. AB - The Quality of Life Scale (QLS(21)) is widely used in clinical trials involving schizophrenia patients. This study aimed to identify a core subset of QLS(21) items that maintains the validity and psychometric properties of the complete version. A parsimonious subset of items from the QLS(21) that can accurately predict the total scale score was sought and evaluated in 133 schizophrenia patients, using the heuristic algorithm for a regression model. Two additional data sets were used for model validation: a subset of 124 patients who participated in the model construction and who completed the QLS(21) 1 year later as well as a new sample of 40 inpatients. Patients were examined with the Positive and Negative Syndrome Scale (PANSS), the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q), the Talbieh Brief Distress Inventory (TBDI), the Extrapyramidal Symptom Rating Scale (ESRS) and the Global Assessment of Functioning Scale (GAF). Using only five QLS items (social initiatives, adequacy, acquaintances, motivation, and time utilization; QLS(5)) as predictors, the correlation was 0.9805 between the predicted and true QLS totals. Two validation samples confirmed this finding. Additional analyses indicate that the QLS(5) exhibited similar performance to the QLS(21) regarding construct validity, test-retest reliability and responsiveness to changes over time. Thus, the five item condensed Quality of Life Scale for schizophrenia maintains the validity of the full QLS, and has the advantage of shorter administration time. Utilization of the revised QLS(5) in routine care and clinical trials may potentially facilitate evaluation of treatment outcomes in schizophrenia. PMID- 15890414 TI - Aspiration in children with CHARGE syndrome. AB - Patients with Coloboma, Heart defect, choanal Atresia, Retarded development, Genitorenal and Ear abnormalities (CHARGE) syndrome have been reported to be at high risk for aspiration and swallowing difficulties. Aspiration has been implicated as the most common cause of mortality in these patients. To date, however, aspiration and swallowing disorders in CHARGE patients have not been independently studied. OBJECTIVE: To determine the prevalence of aspiration and swallowing dysfunction in children with CHARGE syndrome. METHODS: A retrospective chart review of 30 children with CHARGE syndrome was performed. RESULTS: Eighteen (60%) children had aspiration observed on video swallow study (VSS), flexible endoscopic evaluation of swallowing (FEES), and/or had otherwise unexplained bronchiectasis noted on CT scan of the chest. Twenty-four children (80%) had evidence of abnormal swallowing such as laryngeal penetration, dyscoordination, poor bolus mobility, or pooling of secretions. CONCLUSION: Aspiration and swallowing dysfunction are common in children with CHARGE syndrome. Formal evaluation of swallowing function should be part of the standard otolaryngologic examination for these patients. PMID- 15890415 TI - KP-1212/1461, a nucleoside designed for the treatment of HIV by viral mutagenesis. AB - We report the activities of a novel nucleoside analog against HIV. This nucleoside (KP-1212) is not a chain terminator but exerts its antiviral effects via mutagenesis of the viral genome. Serial passaging of HIV in the presence of KP-1212 causes an increase in the mutation rate of the virus leading to viral ablation. HIV strains resistant to KP-1212 have not yet been isolated. Quite to the contrary, virus treated with KP-1212 exhibited an increased sensitivity not only to KP-1212 but also to another nucleoside reverse transcriptase inhibitor (NRTI), zidovudine. HIV strains resistant to other NRTIs (e.g. zidovudine, lamivudine, stavudine, abacavir, etc.) exhibited no cross-resistance towards KP 1212. Multiple assays confirmed that KP-1212 has a favorable (low) genotoxicity profile when compared to some approved antiviral nucleosides. In addition, KP 1212 is not toxic to mitochondria nor does it exhibit any inhibitory effects on mitochondrial DNA synthesis. PMID- 15890416 TI - Binding affinity and capacity of putative adaptor-mediated sorting of a Type I membrane protein in Leishmania mexicana. AB - The membrane-bound acid phosphatase (MBAP), a Type I membrane protein predominantly associated with endosomal/lysosomal structures of Leishmania mexicana promastigotes, contains motifs in its cytosolic COOH-terminal tail ( MEVWRRYMKFKNKQSEAIIV-COOH) akin to tyrosine- and di-leucine-based sorting signals in multicellular organisms. Here, we first show that the COOH-terminal residues IIV of MBAP, but not the Y-residue, are required for endosomal targeting, suggesting specific binding to an adaptor complex at the cell surface. We then determine whether specific binding can be saturated by analysing the efficiency of endosomal targeting for increasing numbers of MBAP molecules per cell. The ratio of the steady-state abundance of wild-type MBAP on the cell surface to MBAP on endosomes increases until the distribution is no longer different from that observed for a mutant MBAP which lacks the IIV-motif or for a glycosylphosphatidylinositol-anchored form, both of which are distributed according to bulk membrane flow. A quantitative analysis of these in vivo results indicates specific binding to a putative adaptor complex with an affinity of about 10-4M to 50,000 sorting sites on the cell surface. PMID- 15890417 TI - Identification of a structural element that is essential for two functions of transcription factor NusG. AB - The transcription factor NusG from Escherichia coli modulates the rate of transcript elongation by RNA polymerase and the efficiency of Rho-dependent transcript termination. It consists of two globular domains with an extra loop extending out of the amino-terminal domain in the position that is occupied by a third globular domain in some NusG homologues. We have tested the role of this appended mini-domain by assaying the elongation and termination enhancement activities of variants. The results show that variants with changes in their sequence do not cause a loss of functions, whereas variants with the deletions of the residues in that domain are much less active for both functions. This finding suggests that the mini-domain serves as a structural element for an interaction rather than as a site for residue-specific contacts. PMID- 15890418 TI - Bridging advanced glycation end product, receptor for advanced glycation end product and nitric oxide with hormonal replacement/estrogen therapy in healthy versus diabetic postmenopausal women: a perspective. AB - Cardiovascular diseases (CVD) are the most significant cause of death in postmenopausal women. The loss of estrogen biosynthesis with advanced age is suggested as one of the major causes of higher CVD in postmenopausal women. While some studies show beneficial effects of estrogen therapy (ET)/hormonal replacement therapy (HRT) in the cardiovascular system of healthy postmenopausal women, similar studies in diabetic counterparts contradict these findings. In particular, ET/HRT in diabetic postmenopausal women results in a seemingly detrimental effect on the cardiovascular system. In this review, the comparative role of estrogens is discussed in the context of CVD in both healthy and diabetic postmenopausal women in regard to the synthesis or expression of proinflammatory molecules like advanced glycation end products (AGEs), receptor for advanced glycation end products (RAGEs), inducible nitric oxide synthases (iNOS) and the anti-inflammatory endothelial nitric oxide synthases (eNOS). The interaction of AGE-RAGE signaling with molecular nitric oxide (NO) may determine the level of reactive oxygen species (ROS) and influence the overall redox status of the vascular microenvironment that may further determine the ultimate outcome of the effects of estrogens on the CVD in healthy versus diabetic women. PMID- 15890419 TI - Cone-beam-CT guided radiation therapy: A model for on-line application. AB - BACKGROUND AND PURPOSE: This paper presents efficient and generalized processes for the clinical application of on-line X-ray volumetric cone-beam CT imaging (XVI) to improve the accuracy of patient set-up in radiation therapy. XVI image guided therapy is illustrated by application to two contrasting sites, intra cranial radiosurgery and prostate radiation therapy, with very different characteristics regarding organ motion, treatment precision, and imaging conditions. PATIENTS AND METHODS: On-line set-up errors are determined in a two step process. First the XVI data is registered to the planning data by matching the machine-isocenter with the planning-isocenter, respectively. The machine isocenter is defined in the XVI data during the reconstruction. The planning isocenter is defined during the planning process in the planning CT data. Set-up errors are then determined from a second registration to remove residual displacements. The accuracy of the entire procedure for on-line set-up error correction was investigated in precision radiosurgery phantom studies. RESULTS: The phantom studies showed that sub-pixel size set-up errors (down to 0.5mm) can be correctly determined and implemented in the radiosurgery environment. XVI is demonstrated to provide quality skull detail enabling precise skull based on-line alignment in radiosurgery. A 'local XVI' technique was found to give encouraging soft-tissue detail in the high-scatter pelvic environment, enabling on-line soft tissue based set-up for prostate treatment. The two-step process for determination of set-up errors was found to be efficient and effective when implemented with a dedicated six panel interface enabling simultaneous visualization on the XVI and planning CT data sets. CONCLUSIONS: XVI has potential to significantly improve the accuracy of radiation treatments. Present image quality is highly encouraging and can enable bony and soft-tissue patient set-up error determination and correction. As with all image guided treatment techniques the development of efficient procedures to utilize on-line data are of paramount importance. PMID- 15890420 TI - Cell cycle effects of topotecan alone and in combination with irradiation. AB - BACKGROUND AND PURPOSE: To elucidate the role of TP53 on differential effects of topoisomerase I inhibitor topotecan (Hycamtin on radiation sensitivity. MATERIALS AND METHODS: Cell cycle distribution and protein expression of TP53, p21(WAF1/CIP1) and cyclin B was studied in CCD32 lung fibroblasts, glioblastoma cell lines U118 (mutant TP53), and U87 (wildtype TP53) after treatment with topotecan (0.05 and 1 microM) and/or ionizing radiation (2 Gy). RESULTS: Cell cycle effects varied with topotecan concentration, resulting in G1 arrest (1 microM), or S/G2/M arrest (0.05 microM), and was modified differentially in fibroblasts and in glioblastoma cells in combination with irradiation. Phosphorylation of TP53 and expression of p21(WAF1/CIP1) was induced by IR and/or topotecan in CCD32 cells, and in U118 cells after topotecan treatment, accompanied by cyclin B degradation. In U87 cells only 1 microM topotecan generated phosphorylation of TP53 and p21(WAF1/CIP1) expression; 0.05 microM caused stabilization of cyclin B. CONCLUSIONS: The antagonistic effect of combined topotecan/irradiation treatment in fibroblasts was most likely due to an immediate radiation induced G1 arrest, but was not observed in p53 wildtype glioblastoma cells. Thus, the impact of TP53 on the topotecan response remains indistinct, and is obviously influenced by other genomic alterations acquired by tumor cells. PMID- 15890421 TI - Acute esophageal toxicity in non-small cell lung cancer patients after high dose conformal radiotherapy. AB - BACKGROUND AND PURPOSE: To correlate acute esophageal toxicity with dosimetric and clinical parameters for non-small cell lung cancer (NSCLC) patients treated with radiotherapy (RT) alone or with chemo-radiotherapy (CRT). PATIENTS AND METHODS: We analyzed the data of 156 patients with medically inoperable or locally advanced NSCLC. Seventy-four patients were irradiated with high dose RT only, 45 patients with sequential CRT (Gemicitabine/Cisplatin) and 37 patients with concurrent CRT (Cisplatin daily 6 mg/m(2)). The radiation dose delivered ranged from 49.5 to 94.5 Gy (2.25-2.75 Gy per fraction) with an overall treatment time of 5-6 weeks. For all patients the maximal acute esophageal toxicity (RTOG/EORTC criteria) was scored and related to dose-volume parameters, as well as to clinical and treatment-related parameters. All parameters were tested univariable and multivariable in a binary logistic regression model. The toxicity data of a homogeneous subgroup was fitted to the Lyman-Kutcher-Burman model. RESULTS: Grade 2 acute esophageal toxicity or higher occurred in 27% (n=42) of the patient population of which nine patients developed grade 3 toxicity and one patient grade 4. All 10 patients with grade>or=3 esophageal toxicity received concurrent CRT. At multivariable analysis, the most significant clinical parameter to predict acute esophageal toxicity was the concurrent use of CRT. The most significant dosimetric parameter was the esophagus volume that received at least 35 Gy. The data of the patients who did not receive concurrent CRT were well described by the Lyman-Kutcher-Burman normal tissue complication probability model. The optimal fit of the data of non-concurrent treated patients to this model was obtained using the following values for the parameters: TD(50)=47 Gy (41-60 Gy), n=0.69 (0.18-6.3) and m=0.36 (0.25-0.55) where the numbers between brackets denote the 95% confidence interval. Acute esophageal toxicity was not significantly increased for patients treated with sequential CRT. CONCLUSION: Both concurrent CRT and the volume that receives at least 35 Gy were predictors of acute esophageal toxicity. PMID- 15890422 TI - Proton beam therapy: too expensive to become true? PMID- 15890423 TI - The impact of hormone therapy on post-implant dosimetry and outcome following Iodine-125 implant monotherapy for localised prostate cancer. AB - BACKGROUND AND PURPOSE: Many patients with localised prostate cancer present with symptoms of benign prostatic hypertrophy (BPH) and require neoadjuvant hormone therapy to shrink the gland before brachytherapy. The impact of this hormone therapy has been evaluated in 667 patients treated with Iodine seed monotherapy. PATIENTS AND METHODS: Prospective data from 667 patients treated between 1995 and 2001 by I-125 seeds prostate implant as monotherapy were analysed. The mean age was 63 years (42--77 years). Three hundred and forty-six (51.9%) patients had a short course of neo adjuvant hormone therapy and 321 (49.1%) did not. The prescribed minimum peripheral dose was 145 Gy (TG 43). Patients were followed up to a maximum of 8.2 years and a minimum of 18 months. Statistical analysis was performed to identify factors that would predict PSA relapse-free survival (PSA RFS) defined by the American Society for Therapeutic Radiology and Oncology (ASTRO). RESULTS: Overall the PSA relapse-free survival is 76.1 and 72.6% for patient cohorts being on pre-treatment hormones and not, respectively (P=0.107). Subdivided into risk groups the low risk group showed 92.5% PSA-RFS with hormones and 75.1% without (P=0.327). The intermediate group 75.7% with hormones and 72.9% without (P=0.148) and for the high-risk group 51.1% with and 51.1% without hormones (P=0.942). Evaluation of post-implant dosimetry in patients with and without hormone therapy showed that the D90 for those who received hormone therapy was 130.8 Gy compared with 145.1 Gy for those who did not (P<0.001). This may be related to the degree of oedema at the time of post-implant dosimetry. The CT to ultrasound prostate volume ratio was 1.17 for patients who received hormone therapy and 0.98 for those who did not (P<0.001). It is suggested that in the interval between stopping hormone therapy and doing post-implant dosimetry there was an increase in prostate volume, which results in a lower D90. Significant correlation was found between D90 and prostate volume on post-implant CT dosimetry with higher D90s for small volume prostates (P<0.001). CONCLUSIONS: Overall hormone therapy had no significant effect on outcome. The apparent lower D90 in hormone treated patients may be related to a change in volume between pre implant and post-implant dosimetry. PMID- 15890424 TI - Cone-beam-CT guided radiation therapy: technical implementation. AB - BACKGROUND AND PURPOSE: X-ray volumetric imaging system (XVI) mounted on a linear accelerator is available for image guidance applications. In preparation for clinical implementation, phantom and patient imaging studies were conducted to determine the irradiation parameters that would trade-off image quality, patient dose and scanning time. PATIENTS AND METHODS: The XVI image quality and imaging dose were benchmarked against those obtained with a helical CT scanner for a head and body phantom. The irradiation parameters were varied including the total imaging dose, number of projections, field of view, reconstruction resolution and use of a scatter rejection grid. We characterized the image quality based on relative contrast, noise, contrast to noise ratio (CNR) and point spread function (PSF). XVI scans of pelvis, head and neck and lung patients were acquired and submitted to a range of observers to identify the favorable reconstruction parameters. RESULTS: Phantom studies have demonstrated that a scatter rejection grid reduces photon scattering and improves the image uniformity. For the body phantom, the helical CT and the wide field XVI technique produce similar image quality, with surface doses of 0.025 and 0.044 Gy respectively. We have demonstrated that the local tomography technique improves the image contrast and the CNR while reducing the skin dose by 40-50% compared to the wide field technique. Clinical scans of head and neck, lung and prostate patients present good soft tissue contrast and excellent bone definition. CONCLUSIONS: With adjustment of irradiation parameters and an imaging surface dose of less than 0.05 Gy, high quality XVI images can be obtained for a phantom simulating the body thickness. XVI is currently feasible for image-guided treatments of head and neck, torso and pelvic areas using soft tissue and bony structures. PMID- 15890425 TI - Evaluation of anatomy-based dwell position and inverse optimization in high-dose rate brachytherapy of prostate cancer: a dosimetric comparison to a conventional cylindrical dwell position, geometric optimization, and dose-point optimization. AB - BACKGROUND AND PURPOSE: To compare treatment planning methods in high-dose-rate (HDR) brachytherapy of prostate cancer. In particular, to assess quantitatively the dosimetric superiority, if any, of the anatomy-based dwell position (ABDP) and inverse optimization (IO) over the conventional cylindrical dwell position (CDP), geometric optimization (GO), and dose-point optimization (DO) in terms of the determination of dwell positions and dwell times. PATIENTS AND METHODS: Between September 2002 and April 2003, 10 cases of treatment-planning CT images were taken for external radiotherapy for prostate cancer. Treatment planning computer software and the CT data were used to create hypothetical HDR brachytherapy applicator needles, which were properly implanted in the prostate. Six different plans including IO with ABDP (IO(ABDP)), IO with CDP (IO(CDP)), GO with ABDP (GO(ABDP)), GO with CDP (GO(CDP)), DO with ABDP (DO(ABDP)), and DO with CDP (DO(CDP)) were made for each case, that is, 60 plans in total. All plans were normalized so that the D(95) should be equal to 100% of the prescribed dose. Dose volume histograms from all 60 plans were analyzed, and multiple implant quality indices, including CI, EI, DNR, %V(R 75), %V(B 75), and %V(U 150) for each plan, were extracted and compared. Then, the best settings for IO(ABDP) regarding dwell position and dose limit were sought for. RESULTS: ABDP showed a statistically significantly lower EI (P<0.001), %V(R 75) (P=0.002), and %V(B 75) (P=0.015) than CDP. IO showed a statistically significantly lower %V(U 150) than GO (P=0.009), or than DO (P<0.001). Given a definition that a figure exceeding three-fold of the minimum figure of the index is clinically unacceptable, only in IO(ABDP) all index figures were clinically acceptable, while in the other five plans at least one index figure was unacceptable. CONCLUSIONS: In the CT-based treatment planning for prostate HDR brachytherapy, ABDP is useful to achieve a high conformity, which leads to a reduction of the doses to the bladder, rectum, and surrounding normal tissue. IO is useful to lower the urethral dose without sacrificing conformity. IO(ABDP) is recommended on the basis of the current study. However, this conclusion has been drawn from the idealized hypothetical settings, and some possibility remains that this conclusion is not always applicable to the real implants. PMID- 15890426 TI - A multiwavelength fluorescence probe: is one probe capable for on-line monitoring of recombinant protein production and biomass activity? AB - Monitoring cell culture performance requires maximizing the number and the quality of measured parameters and in situ 2D fluorescence spectroscopy could allow intensification of simultaneous data acquisition. The use of a multiwavelength fluorescence probe is proposed for monitoring GFP-producing cultures in bioreactor. The yeast Pichia pastoris and NSO mammalian cells were studied as model systems. Tryptophan, NAD(P)H and riboflavins (riboflavin, FMN, FAD) signals were effective for on-line yeast biomass estimation during the growth phase. During the GFP production phase, in situ measurements of the GFP concentration from the fluorescence probe were well correlated with off-line analyses. Tryptophan and NAD(P)H signals diverged from that of biomass during GFP production. With NSO mammalian cells, results showed that the culture parameters have to be optimized for the use of a fluorescence probe. The use of serum and phenol-red interfered with NAD(P)H and riboflavins fluorescence signals. Nevertheless, it appears that a multiwavelength probe could be useful for culture monitoring of biomass, cell activity and recombinant protein expression in an optimized culture medium. PMID- 15890427 TI - Prevalence and control of dyslipidemia among persons with diabetes in the United States. AB - OBJECTIVE: We assessed the prevalence, treatment, and control of dyslipidemia among United States (U.S.) adults with diabetes. METHODS: Among 498 adults (projected to 13.4 million) aged >or=18 years with diabetes representative of the U.S. population and surveyed within the cross-sectional National Health and Nutrition Examination Survey 1999-2000, control of lipids was classified according to American Diabetes Association criteria. The extent of low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), and triglyceride (TG) control was examined by gender and ethnicity, in comparison to those without diabetes, and according to lipid-lowering treatment. Analyses were weighted to the U.S. population. RESULTS: Less than one-third of men and only one-fifth of women with diabetes are in control for LDL-C, defined as <2.6 mmol/l (<100mg/dl); over 70% are not at goal. Over half of men and over two thirds of women have low levels of HDL-C (or=1.7 mmol/l [150 mg/dl]). Low HDL-C was more common in Caucasians (70.1%) than in Hispanics (58.8%) or African-Americans (41.5%) (p<0.001). 28.2% of subjects with diabetes were on lipid-lowering treatment. Control of LDL-C did not differ by treatment status and only 3% of subjects were controlled to target levels for all lipids. CONCLUSION: Many persons with diabetes remain uncontrolled for dyslipidemia. Intensified efforts at screening and treatment according to current guidelines are warranted. PMID- 15890428 TI - Glycemic control and morbidity in the Canadian primary care setting (results of the diabetes in Canada evaluation study). AB - The objective of this national, cross-sectional study was to provide insight into the care and treatment of type 2 diabetes (T2DM) in the Canadian primary care setting. Specifically, the study examines glycemic control, management and morbidity load among T2DM patients, and investigates the relationship of glycemic control and morbidity load with duration of diabetes. Participating primary care physicians (PCPs) (N=243) completed a chart audit for the first 10 patients with T2DM attending their clinics (2473 eligible patient records). The mean A1C was 7.3% with 49% of patients not at target (A1C>or=7.0%). Glycemic control eroded significantly with increasing duration of diabetes in spite of increasing therapeutic intervention. T2DM patients experienced a high morbidity load (hypertension 63%; dyslipidemia 59%; macrovascular complications 28%; microvascular complications 38%) each of which increased significantly with duration of diabetes. For 79% of patients not at target, PCPs identified lifestyle intervention as the strategy for achieving glycemic targets while more aggressive treatment plans were identified for only 56%. These results underscore the complexity of primary care management of T2DM and suggest that current treatment approaches are not intensive enough for a large proportion of patients especially those with longer duration of disease. PMID- 15890429 TI - Comparison of the quality of diabetes care in primary care diabetic clinics and general practice clinics. AB - OBJECTIVES: To compare the quality of care provided to diabetic hypertensive patients by diabetic clinics versus general practice clinics (GP-clinics) in a primary care setting. MATERIALS AND METHODS: A retrospective analysis of medical records of patients from six primary care centres in Bahrain. RESULTS: The recommended target blood pressure <130/85 mmHg was achieved in 6.8 and 10%, and glycated haemoglobin <7% was attained in 14.8 and 11% of patients attending diabetic clinics (n=177) and GP-clinics (n=180), respectively. Although complementary antihypertensive combinations were prescribed at a rate less than that recommended in guidelines, combinations were significantly more often prescribed for patients attending the GP-clinics (46.7% [95% CI 39.4-53.9] versus 33.4% [CI 26.8-40.6]; p=0.01). The prescribing pattern and rank-order of antihypertensive and oral hypoglycaemic agents prescribed, either as monotherapy or in combinations, were similar in both clinics. The majority of diabetic hypertensive patients attending diabetic clinics or GP-clinics were at high cardiovascular risk. Anthropometric characteristics were recorded consistently in patients attending diabetic clinics. Laboratory investigations were extensively utilized for monitoring patients attending diabetic clinics. CONCLUSIONS: In primary care, in both diabetic clinics and GP-clinics, hypertension and diabetes were inadequately controlled, with no difference between the two settings. An integrated approach involving diabetic clinics managed by primary care physicians and nurses trained in diabetes management, and supported by national guidelines, is needed. PMID- 15890430 TI - Ascites from cirrhotic patients induces angiogenesis through the phosphoinositide 3-kinase/Akt signaling pathway. AB - BACKGROUND/AIMS: Ascites in patients with cirrhosis is associated with worsening of systemic hemodynamics. Thus, the aim of this study was to investigate the biological activity of ascites on endothelial cells. METHODS: Human umbilical vein endothelial cells (HUVECs) were used to investigate the angiogenic activity of ascites obtained from cirrhotic patients. RESULTS: Ascites-induced Akt activation, cell migration and tube formation in HUVECs. The pretreatment of HUVECs with the phosphatidylinositide 3-kinase (PI3-kinase) inhibitor LY294002, resulted in a decrease in chemotaxis and cell tube formation induced by ascites. Moreover, the inhibition of Akt activity in HUVECs by transduction of an inactive phosphorylation Akt mutant (AA-Akt), blocked tube formation. These angiogenic effects of ascites were also operative in vivo showing a PI3-kinase activation dependence in the angiogenesis induced by ascites. In addition, the preincubation of ascites with anti-fibronectin antibody led to a significant decrease in HUVECs migration, cell tube formation and in vivo angiogenesis. CONCLUSIONS: These results confirm the novel concept that ascites is a bioactive fluid which can modify vascular properties through the activation of the PI3-kinase/Akt pathway in endothelial cells. Furthermore, our results demonstrated that this ascites induced mechanism is mediated, at least in part, by fibronectin. PMID- 15890431 TI - Changes in antibiotic resistance of the most common Gram-negative bacteria isolated in intensive care units. AB - We studied the changes in antibiotic resistance of the most common Gram-negative bacteria isolated in the intensive care units at our hospital in 2000 and 2002. Bacterial identification was performed by use of the VITEK 60 analyser, and antibiotic susceptibilities were tested by the VITEK 60 analyser and the disk diffusion agar method. The bacteria isolated most frequently were Pseudomonas aeruginosa (132 strains in 2000 and 106 in 2002), Acinetobacter calcoaceticus (98 and 109 strains, respectively) and Klebsiella pneumoniae (53 and 83 strains, respectively). Acinetobacters presented the highest percentage resistance, with significant increases in resistance to imipenem (15% in 2000 and 67% in 2002) and piperacillin/tazobactam (41% and 72%, respectively). P. aeruginosa presented a significant increase in resistance to all antibiotics, except ceftazidime. A large increase was observed in the resistance of K. pneumoniae to amikacin (from 10% to 50%), ceftazidime (from 80% to 90%) and tobramycin (from 80% to 90%). No imipenem-resistant strains of K. pneumoniae were found. PMID- 15890432 TI - Sterilization breach. PMID- 15890433 TI - A change in Belgian epidemic methicillin-resistant Staphylococcus aureus phage types in 2000: phenotypic and genotypic characterization of isolates from a general hospital. AB - During 2000, new methicillin-resistant Staphylococcus aureus (MRSA) epidemic phage types became preponderant in Belgium. In the present study, phenotypic and genotypic characteristics of 130 MRSA isolates from a general Belgian hospital were investigated. The MRSA nature of the isolates was confirmed by coagulase test, oxacillin screen plate test and detection of the mecA gene by polymerase chain reaction. Phage typing categorized the MRSA strains into two main groups: the [O]* types and the [J]* types. SmaI macrorestriction analysis by pulsed-field gel electrophoresis gave the same pulsotype in the majority of strains. All strains of the [O]* and [J]* groups, except one, belonged to this pulsotype. Aminoglycoside-modifying-enzyme genes could only be detected in a minority of strains. Although the epidemic phage types of the mid-1990s appear to have been supplanted by the [O]* and [J]* groups, the MRSA population examined showed a remarkably uniform profile corresponding to the previous major clone B. PMID- 15890434 TI - Ammonia effects on pyruvate/lactate production in astrocytes--interaction with glutamate. AB - Ammonia exerts a multitude of metabolic and non-metabolic effects on brain tissue. In the present communication we have investigated its effect on lactate production rates, pyruvate production rates and pyruvate/lactate ratios in mouse cerebrocortical astrocytes and neurons in primary cultures. No effects were found in neurons. All three parameters were affected by ammonia in astrocytes, but less potently and to a smaller degree in cells that had been treated with dibutyryl cyclic AMP (morphologically differentiated cells) than in untreated cells (morphologically undifferentiated cells). In the differentiated cells ammonia had virtually no effect up to a concentration of 1.0 mM, but at 3.0 mM it increased lactate production and decreased pyruvate/lactate ratio significantly. In the undifferentiated cells ammonia greatly increased lactate accumulation (by 80% at 3.0 mM) and it inhibited pyruvate accumulation (by 40% at 3.0 mM). It thereby reduced the pyruvate/lactate ratio progressively within the entire range 0.1-3.0 mM ammonia. In support of the hypothesis that the ammonia-induced reduction of pyruvate/lactate ratio is secondary to depletion of cellular glutamate by formation of glutamine (and glutathione) and a resulting interruption of the malate-aspartate shuttle (MAS), the addition of glutamate to the incubation medium significantly diminished the ammonia-induced reduction of pyruvate/lactate ratio, whereas it had no effect on the increased lactate production. It is discussed that MAS interruption may have additional consequences in astrocytes. PMID- 15890435 TI - mRNA up-regulation of MHC II and pivotal pro-inflammatory genes in normal brain aging. AB - In normal brain aging, CNS resident macrophages exhibit increased expression of major histocompatibility complex (MHC) II expression. However, the transcriptional basis for this observation has not been clarified nor have age related alterations in pivotal pro-inflammatory genes been characterized. Age related mRNA alterations in MHC II, MHC II accessory molecules and several pro inflammatory mediators were measured in older (24 months) and younger (3 months) male F344xBN F1 rats. Real time RT-PCR was utilized to measure steady state mRNA levels in hippocampus. Older as compared to younger animals exhibited increased mRNA levels of MHC II, CD86, CIITA and IFN-gamma. Furthermore, IL-10 and CD200 mRNA, molecules that down-regulate macrophage activation, was decreased in older animals. The present results indicate that normal brain aging is characterized by a shift towards a pro-inflammatory microenvironment in the CNS. PMID- 15890436 TI - Novel pyrazinone and pyridinone thrombin inhibitors incorporating weakly basic heterobicyclic P(1)-arginine mimetics. AB - The design, synthesis and biological activity of new thrombin inhibitors with a pyridinone or pyrazinone core and different heterobicyclic P(1) arginine side chain mimetics are described. The arginine side-chain mimetics used in this study are (+/-)-4,5,6,7-tetrahydro-2H-indazol-5-ylmethanamine and both enantiomers thereof, (+/-)-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine and the corresponding R enantiomer. Compound 25, the most potent in the series of pyrazinone inhibitors, exhibited a K(i) of 41 nM in vitro and high selectivity against trypsin and factor Xa. PMID- 15890437 TI - Ergonomic identification and biomechanical evaluation of workers' strategies and their validation in a training situation: summary of research. AB - Our goal was to evidence the role of workers in the search for safer manual handling strategies. Different strategies used by expert and novice workers, such as footwork (positioning/displacement), box manoeuvres (handgrips and load tilting) and posture were ergonomically identified. They were biomechanically evaluated to shed light on their potential for safe handling by reducing back loadings, back asymmetries and mechanical work requirements. The experts' strategies were validated in a training situation and evaluated by two independent studies among novice workers. The first, a control study, showed that free practice did not lead to safer handling practices. The second, a training situation based on observing contrasted performances of experts and novices for footwork, box manoeuvres and back posture, when combined with practice and the search for optimal solutions, prompted the novices to adopt new ways of manoeuvring boxes and positioning themselves that appear safer for back efforts, asymmetries, and mechanical work. These elements should be included in educational programs for safe handling. RELEVANCE: The observation of contrasted strategies of expert and novice workers using an ergonomic approach, supplemented by biomechanical evaluations of these strategies, is a key factor in designing training programs for safe handling. PMID- 15890438 TI - Age-dependent differences in lateral balance recovery through protective stepping. AB - BACKGROUND: Aging appears to present particular problems for lateral balance stability related to falls. Protective stepping is a common strategy for maintaining balance that may be impaired with aging due to changes in neuromusculoskeletal factors. This study assessed the response patterns, kinematics, and single support hip abduction torque during lateral protective stepping for balance recovery in healthy young and elderly adults. METHODS: Ten healthy elderly and 10 younger adults received stepper-motor driven waist-pulls of bipedal stance applied pseudorandomly to either side. Stepping response strategies were quantified with force platforms and motion analysis. FINDINGS: The young responded primarily using a single lateral sidestep with the limb that was initially loaded passively by the waist-pull, while older subjects favored crossover stepping using multiple steps with more inter-limb collisions. When the elderly did use loaded side steps, the steps were longer, slower, and higher and included greater and prolonged lateral trunk motion than in the young. Overall, older subjects produced greater and less rapid stabilizing hip abduction torque during the single support phase. INTERPRETATION: Age-associated differences in lateral balance control through stepping included using a riskier recovery strategy with increased collisions between the limbs, multiple steps, altered first step characteristics and lateral trunk motion during direct sidestepping, and a generally greater support hip torque. The difficulties with lateral balance control in aging may reflect factors such as impaired hip abduction torque-time capacity and lateral trunk mobility/control. Our findings contribute additional knowledge pertaining to the problem of balance dysfunction and falls among the elderly. PMID- 15890439 TI - Requirements for upper extremity motions during activities of daily living. AB - BACKGROUND: Functioning of the upper extremity after implantation of an endoprosthesis remains limited despite the achieved pain relief. Upper extremity kinematics can give insight into function after shoulder arthroplasty. Data on ranges of motion related to the performance of a selection of activities of daily living can aid the clinician in evaluating the outcome of the shoulder and elbow arthroplasties. METHODS: Cross-sectional descriptive study of range of motion and activities of daily living kinematics, conducted on non-impaired subjects. The shoulder and elbow motions of 24 healthy female subjects are measured with an electromagnetic tracking device while performing 8 range of motion tasks and five activities of daily living. The angles of shoulder and elbow are calculated during these tasks. RESULTS: A data set with upper extremity joint angles has been obtained. Large glenohumeral rotations are found for the tasks that require high elevation angles. Large axial rotations of the humerus are found for two of the activities of daily living tasks: the perineal care task and the hair-combing task. Large elbow flexions were seen in the following tasks: combing hair, washing the axilla and eating with a spoon. INTERPRETATION: This study shows a suitable method to describe range of motion and activities of daily living and can serve as a starting point for developing a database on how activities of daily living are performed in a larger population and which joint angles are required to perform these tasks. Results can be used to identify restrictions in upper extremity functioning in patients with shoulder impairments. PMID- 15890440 TI - Effects of nitrogen enrichment on coastal dune grassland: a mesocosm study. AB - Mesocosms filled with dune sand were planted with graminoid (Calamagrostis epigejos, Carex arenaria) and herbaceous species (Carlina vulgaris, Galium verum). Strong effects of nitrogen addition on the vegetation were found within two to three years. The above-ground biomass of C. epigejos and C. arenaria increased at deposition rates between 10 and 80 kg N ha(-1) yr(-1). Both grasses were limited by N. In latter stages P limitation was suggested for C. arenaria. At high N-levels, C. epigejos dominated the vegetation within two years. C. vulgaris and G. verum declined drastically as a result of increased competition for light by the highly competitive grass C. epigejos. It is concluded that increased (ambient) N inputs are of major importance for the increased dominance of tall grasses in stable dune grasslands. PMID- 15890441 TI - Fluoroquinolones cause changes in extracellular matrix, signalling proteins, metalloproteinases and caspase-3 in cultured human tendon cells. AB - Antimicrobial therapy with fluoroquinolones can be associated with tendinitis and other tendon disorders as an adverse reaction associated with this class of antimicrobials. Here we investigated aspects of the mechanism of quinolone induced tendotoxicity in human tenocytes focussing mainly on the question whether fluoroquinolones may induce apoptosis. Monolayers of human tenocytes were incubated with ciprofloxacin or levofloxacin at different concentrations (0, 3, 10, 30 and 100mg/L medium) for up to 4 days. Ultrastructural changes were studied by electron microscopy, and alterations in synthesis of specific proteins were determined using immunoblotting. At concentrations, which are achievable during quinolone therapy, 3mg ciprofloxacin/L medium significantly decreased type I collagen; similar changes were observed with 3mg ciprofloxacin or 10mg levofloxacin/L medium for the beta(1)- integrin receptors. Effects were intensified at higher concentrations and longer incubation periods. Cytoskeletal and signalling proteins, such as activated shc or erk 1/2, were significantly reduced by both fluoroquinolones already at 3mg/L. Furthermore, time- and concentration-dependent increases of matrix metalloproteinases as well as of the apoptosis marker activated caspase-3 were found. Apoptotic changes were confirmed by electron microscopy: both fluoroquinolones caused typical alterations like condensed material in the nucleus, swollen cell organelles, apoptotic bodies and bleb formation at the cell membrane. Our results provide evidence that besides changes in receptor and signalling proteins apoptosis has to be considered as a final event in the pathogenesis of fluoroquinolone-induced tendopathies. PMID- 15890442 TI - Paraplegia due to spinal subdural hematoma as a complication of posterior fossa surgery: Case report and review of the literature. AB - Although blood contamination of cerebrospinal fluid (CSF) after an intracranial operation is possible, development of a symptomatic spinal hematoma after a posterior fossa surgery has never been reported. A 43-year-old woman underwent a posterior fossa tumor removal in the prone position with no intraoperative difficulty. On the second postoperative day, she complained of severe epigastric pain and developed a rapid onset of paraplegia with anesthesia below the thoracic 5 spinal level. The emergency cranial and spinal MRIs revealed a spinal extramedullary hemorrhage spreading to the whole spinal regions, just sparing the cauda equina area. There was a prominent localized hematoma formation surrounding and compressing the spinal cord at the upper thoracic levels, which was evacuated via an urgent laminectomy. The patient showed partial neurological recovery after the decompression. Development of the spinal hematoma was explained by the movement of blood from the tumor bed into the spinal canal under the effect of gravity, during or after the operation. A 30 degrees head elevation might facilitate the accumulation of blood. Localization of the hematoma formation may be caused by the fact that the upper thoracic levels constitute the apex of the kyphosis. We conclusively suggest that a spinal hematoma should be taken into consideration as a rare but potentially severe complication of a posterior fossa surgery. Meticulous hemostasis and isolation of the surgical area from the spinal spaces are essential. Overdrainage of CSF should be abandoned. Postoperatively, patients should be monitored for spinal findings as well as cranial signs. PMID- 15890443 TI - Bilateral carotid-cavernous fistula following head trauma: Possible worsening of brain injury following balloon catheter occlusion? AB - A 68-year-old woman developed right pulsatile exophtalmos in the early course of facial and head trauma. Investigations by visual evoked potentials suggested the presence of a bilateral prechiasmatic lesion. Intraocular pressure rapidly increased and a high-flow carotid-cavernous fistula was evident at carotid angiography. Immediately after the treatment of the right fistula by balloon embolisation, a systolic bruit was noted over the left eyeball and angiography disclosed the presence of a left carotid-cavernous fistula that was treated by the same approach. The patient developed brain oedema leading to death and we postulated that reestablishment of normal cerebral perfusion after abrupt closure of the fistulas may have played a deleterious role. PMID- 15890444 TI - Inhibition of protein phosphatase 2B upregulates serine phosphorylation of N methyl-D-aspartate receptor NR1 subunits in striatal neurons in vivo. AB - This study investigated the role of protein phosphatase 2B (calcineurin) in regulating phosphorylation of N-methyl-D-aspartate receptor (NMDAR) NR1 subunits and other phosphoproteins in the rat striatum in vivo. In chronically cannulated rats, microinjection of the calcineurin selective inhibitor cyclosporin A increased phosphorylation of NMDAR NR1 subunits at serine 896 and serine 897 in the injected dorsal striatum. The increase in NMDAR NR1 phosphorylation was dose dependent in a dose range surveyed (0.005, 0.05, and 0.5 nmol). Parallel with increased serine phosphorylation of NR1 subunits, cyclosporin A dose-dependently increased phosphorylation of a Ca2+-sensitive protein kinase, extracellular signal-regulated protein kinase 1/2 (ERK1/2), and a Ca2+/cAMP-sensitive transcription factor, cAMP response element-binding protein (CREB), in the dorsal striatum. Using an immediate early gene product Fos as a reporter of inducible gene expression, cyclosporin A was found to upregulate Fos expression in the dorsal striatum. These results indicate that calcineurin plays an important role in the tonic dephosphorylation of NMDAR NR1 subunits and other two key cytoplasmic and nuclear signaling proteins (ERK1/2 and CREB) in striatal neurons. PMID- 15890445 TI - Active pixel sensor array as a detector for electron microscopy. AB - A new high-resolution recording device for transmission electron microscopy (TEM) is urgently needed. Neither film nor CCD cameras are systems that allow for efficient 3-D high-resolution particle reconstruction. We tested an active pixel sensor (APS) array as a replacement device at 200, 300, and 400 keV using a JEOL JEM-2000 FX II and a JEM-4000 EX electron microscope. For this experiment, we used an APS prototype with an area of 64 x 64 pixels of 20 microm x 20 microm pixel pitch. Single-electron events were measured by using very low beam intensity. The histogram of the incident electron energy deposited in the sensor shows a Landau distribution at low energies, as well as unexpected events at higher absorbed energies. After careful study, we concluded that backscattering in the silicon substrate and re-entering the sensitive epitaxial layer a second time with much lower speed caused the unexpected events. Exhaustive simulation experiments confirmed the existence of these back-scattered electrons. For the APS to be usable, the back-scattered electron events must be eliminated, perhaps by thinning the substrate to less than 30 microm. By using experimental data taken with an APS chip with a standard silicon substrate (300 microm) and adjusting the results to take into account the effect of a thinned silicon substrate (30 microm), we found an estimate of the signal-to-noise ratio for a back-thinned detector in the energy range of 200-400 keV was about 10:1 and an estimate for the spatial resolution was about 10 microm. PMID- 15890446 TI - Azadirachtin-impregnated traps for the control of Dermanyssus gallinae. AB - The effect of neem oil (azadirachtin), originating from the tree Azadirachta indica, was investigated as a potential compound to control the poultry red mite, Dermanyssus gallinae. In vitro tests were performed to determine the most appropriate formulation of neem extracts and concentration of the substance to be used. Cardboard traps containing 20% neem oil were placed at the mites' aggregation sites, out of reach of the hens, in a floor system for layers containing approximately 2400 birds. Treated traps were replaced every week for 4 weeks. Throughout the study, the parasite population was monitored by collections of mites with untreated plastic traps. A 92% reduction of D. gallinae was recorded. PMID- 15890447 TI - Heartworm disease in dogs: unusual clinical cases. AB - Two unusual cases of heartworm infection are described, including one of spontaneous rupture of cardiac chambers, most likely due to the progressive increase in pressure within the right ventricle caused by Dirofilaria immitis infection. Another case report describes persistent left cranial vena cava, which interfered with the surgical removal of heartworm via the left jugular vein. PMID- 15890448 TI - Comparative study of the efficacy of eprinomectin versus ivermectin, and field efficacy of eprinomectin only, for the treatment of chorioptic mange in alpacas. AB - The efficacy of eprinomectin versus ivermectin (Study 1: a single-centre, randomised, treatment-controlled, blinded field trial), and the field efficacy of eprinomectin (Study 2: a single-centre, open, un-controlled field trial) for the treatment of chorioptic infestation in naturally infested alpacas were assessed in two studies. Thirty alpacas, all positive for Chorioptes sp. mite, were randomly allocated to two treatment groups in Study 1. Group A received a single topical administration of a 0.5% formulation of eprinomectin at the dose rate of 500mug/kg. Group B received three subcutaneous administrations at 14 days interval of a 1% formulation of ivermectin at the dose rate of 400mug/kg. Response to treatment was assessed by periodic mite count, and skin lesions scored. In Study 2, one group of 19 alpacas received four administrations at weekly interval of topical eprinomectin at the dose rate of 500mug/kg, and response to treatment was monitored by mite counts. No localised or systemic side effects were observed in either trial. There was a statistically significant decrease in mite counts on day 7 (P<0.001) within treatment Groups A and B of Study 1, but mite counts increased again on day 14 and remained high for the duration of the trial in both treatment groups. On day 14 of Study 2, there was a statistically significant reduction in mite counts (P<0.008) and the mite counts remained very low throughout the remainder of the study. The eprinomectin protocol employed in Study 2, consisting of four weekly topical administrations at the dose rate of 500mug/kg of body weight, proved highly effective at reducing the Chorioptes mite burden in alpacas. PMID- 15890449 TI - Mechanism of neopterin-induced myocardial dysfunction in the isolated perfused rat heart. AB - Neopterin is a sensitive marker for diseases involving increased activity of the cellular immune system in humans. Many studies, however, provide evidence for neopterin not only as a marker, but also for its characteristic effects. Recently, we were able to demonstrate a considerable influence of exogenous neopterin at a concentration of 100 mumol/l on cardiac performance in the Langendorff model of isolated perfused rat hearts. The present study was designed to investigate its possible mechanism. During co-infusion of neopterin at a concentration of 100 mumol/l with the unspecific nitric oxide synthase inhibitor N(G)-monomethyl-l-arginine monoacetate, the nitric oxide donor PAPA NONOate, the free radical scavenger N-acetylcysteine, or the pro-inflammatory cytokine tumor necrosis factor-alpha the effects on cardiac contractility parameters and coronary vascular resistance were studied in 67 male Sprague-Dawley rats. The temperature-controlled and pressure-constant Langendorff apparatus was used with retrograde perfusion of the aorta and a Krebs-Henseleit buffer. Neither the unspecific nitric oxide synthase inhibitor nor the nitric oxide donor excludes nitric oxide from playing a mechanistic role in our perfusion studies. Tumor necrosis factor-alpha was without any synergistic or antagonistic effects when co treated with neopterin. N-acetylcysteine was most effective in abolishing neopterin-dependent effects on cardiac function. The negative effects of neopterin on cardiac performance might be due to an enhancement of oxidative stress by neopterin that can be attenuated by the antioxidant N-acetylcysteine. Neopterin has to be considered a pathogenic factor in the development of cardiac dysfunction in chronic disease states with high neopterin levels secondary to activation of the immune system. PMID- 15890450 TI - Activation of a GST-tagged AKT2/PKBbeta. AB - The protein kinase AKT is a key regulator for cell growth, cell survival and metabolic insulin action. However, the mechanism of activation of AKT in vivo, which presumably involves membrane recruitment of the kinase, oligomerization, and multiple phosphorylation events, is not fully understood. In the present study, we have expressed and purified dimeric GST-fusion proteins of human protein kinase AKT2 (DeltaPH-AKT2) in milligram quantities via the baculovirus expression system. Treatment of virus-infected insect cells with the phosphatase inhibitor okadaic acid (OA) led to phosphorylation of the two regulatory phosphorylation sites, Thr309 and Ser474, and to activation of the kinase. Likewise, phosphorylation of Thr309 in vitro by recombinant PDK1 or mutation of Thr309 and Ser474 to acidic residues rendered the kinase constitutively active. However, even though the specific activity of our AKT2 was increased 15-fold compared to previous reports, GST-mediated dimerization alone did not lead to an activation of the kinase. Whereas both mutagenesis and phosphorylation led to an increase in the turnover number of the enzyme, only the latter resulted in a marked reduction (20-fold) of the apparent Km value for the exogenous substrate Crosstide, indicating that this widely used mutagenesis only partially mimics phosphorylation. Kinetic analysis of GST-AKT2 demonstrates that phosphorylation of Thr309 in the activation loop of the kinase is largely responsible for the observed reduction in Km and for a subsequent 150-fold increase in the catalytic efficiency (k(cat)/Km) of the enzyme. Highly active AKT2 constructs were used in autophosphorylation reactions in vitro, where inactive AKT2 kinases served as substrates. As a matter of fact, we found evidence for a minor autophosphorylation activity of AKT2 but no significant autophosphorylation of any of the two regulatory sites, Thr309 or Ser474. PMID- 15890451 TI - Effects of a time-varying strong magnetic field on transient increase in Ca2+ release induced by cytosolic Ca2+ in cultured pheochromocytoma cells. AB - Exposure of pheochromocytoma (PC 12) cells to a time-varying 1.51 T magnetic field inhibited an increase in the intracellular Ca2+ concentration ([Ca2+]i) induced by addition of caffeine to Ca(2+)-free medium. This inhibition occurred after a 15-min exposure and was maintained for at least 2 h. [Ca2+]i sharply increased in cells loaded with cyclic ADP-ribose, and 2-h exposure significantly suppressed the increase. Addition of ATP induced a transient increase in intracellular Ca2+ release mediated by IP3 receptor, and this increase was strongly inhibited by the exposure. Results indicated that the magnetic field exposure strongly inhibited Ca2+ release mediated by both IP3 and ryanodine receptors in PC 12 cells. However, thapsigargin-induced Ca2+ influx (capacitative Ca2+ entry) across the cell membrane was unaffected. The ATP content was maintained at the normal level during the 2-h exposure, suggesting that ATP hydrolysis was unchanged. Therefore, Mg2+ which is known to be released by ATP hydrolysis and inhibit intracellular Ca2+ release may not relate the exposure caused inhibition. Eddy currents induced in culture medium appear to change cell membrane properties and indirectly inhibit Ca2+ release from endoplasmic reticulum and other Ca2+ stores in PC 12 cells. PMID- 15890452 TI - Influence of external pH on two types of low-voltage-activated calcium currents in primary sensory neurons of rats. AB - The influence of extracellular pH (pH(o)) on low-voltage-activated calcium channels of acutely isolated DRG neurons of rats was examined using the whole cell patch-clamp technique. It has been found that in the neurons of middle size with capacitance C=60+/-4.8 pF (mean+/-S.E., n=8) extracellular acidification from pH(o) 7.35 to pH(o) 6.0 significantly and reversibly decreased LVA calcium current densities by 75+/-3.7%, shifted potential for half-maximal activation to more positive voltages by 18.7+/-0.6 mV with significant reduction of its voltage dependence. The half-maximal potential of steady-state inactivation shifted to more positive voltages by 12.1+/-1.7 mV (n=8) and also became less voltage dependent. Dose-response curves for the dependence of maximum values of LVA currents on external pH in neurons of middle size have midpoint pK(a)=6.6+/-0.02 and hill coefficient h=0.94+/-0.04 (n=5). In small cells with capacitance C=26+/ 3.6 pF (n=5), acidosis decreased LVA calcium current densities only by 15.3+/ 1.3% and shifted potential for half-maximal activation by 5.5+/-1.0 mV with reduction of its voltage dependence. Half-maximal potential of steady-state inactivation shifted to more positive voltages by 10+/-1.6 mV (n=4) and also became less voltage dependent. Dose-response curves for the dependence of maximum values of LVA currents on external pH in neurons of small size have midpoint pK(a)=7.9+/-0.04 and hill coefficient h=0.25+/-0.1 (n=4). These two identified types of LVA currents besides different pH sensitivity demonstrated different kinetic properties. The deactivation of LVA currents with weak pH sensitivity after switching off depolarization to -30 mV had substantially longer decay time than do currents with strong pH sensitivity (tau(d) approximately 5 ms vs. 2 ms respectively). It was found that the prolongation of depolarization steps slows the subsequent deactivation of T-type currents in small DRG neurons. Deactivation traces in these neurons were better described by the sum of two exponentials. Thus, we suppose that T-type channels in small DRG neurons are presented mostly by alpha1I subunit. We suggest that these two types of LVA calcium channels with different sensitivity to external pH can be differently involved in the origin of neuropathic changes. PMID- 15890453 TI - Insufficient evidence to compare clinical effectiveness and safety of self expanding metal stents with other treatments for malignant colorectal obstruction. PMID- 15890455 TI - Differential effects of the substrate inhibitor l-trans-pyrrolidine-2,4 dicarboxylate (PDC) and the non-substrate inhibitor DL-threo-beta benzyloxyaspartate (DL-TBOA) of glutamate transporters on neuronal damage and extracellular amino acid levels in rat brain in vivo. AB - The extracellular concentration of glutamate is highly regulated by transporter proteins, due to its neurotoxic properties. Dysfunction or reverse activation of these transporters is related to the extracellular accumulation of excitatory amino acids and neuronal damage associated with ischemia and hypoglycemia. We have investigated by microdialysis the effects of the substrate and the non substrate inhibitors of glutamate transporters, l-trans-2,4-pyrrolidine dicarboxylate (PDC) and DL-threo-beta-benzyloxyaspartate (DL-TBOA), respectively, on the extracellular levels of amino acids in the rat hippocampus in vivo. In addition, we have studied the effect of both inhibitors on neuronal damage after direct administration into the hippocampus and striatum. Electroencephalographic activity was recorded after the intrahippocampal infusion of DL-TBOA or PDC. Microdialysis administration of 500 microM DL-TBOA into the hippocampus increased 3.4- and nine-fold the extracellular levels of aspartate and glutamate, respectively. Upon stereotaxic administration it induced neuronal damage dose dependently in CA1 and dentate gyrus, and convulsive behavior. Electroencephalographic recording showed the appearance of limbic seizures in the hippocampus after DL-TBOA infusion. In the striatum it also induced dose dependent neuronal damage. These effects were prevented by the i.p. administration of the glutamate receptor antagonists (+)-5-methyl-10,11-dihydroxy 5H-dibenzo(a,d)cyclohepten-5,10-iminemaleate and 2,3-dihydroxy-6-nitro-7 sulfamoyl-benzo(F)-quinoxaline. In contrast to dl-TBOA, PDC (500 microM) induced a more discrete elevation of excitatory amino acids levels (2.6- and three-fold in aspartate and glutamate, respectively), no neuronal damage or behavioral changes, and no alterations in electroencephalographic activity. The differential results obtained with DL-TBOA and PDC might be attributed to their distinct effects on the extracellular concentration of amino acids. Results are relevant to the understanding of the role of glutamate transporters in amino acid removal or release and the induction of excitotoxic cell death. PMID- 15890456 TI - Transcription factor gene expression profiling after acute intermittent nicotine treatment in the rat cerebral cortex. AB - Several studies in different in vitro and in vivo models have demonstrated neuroprotective effects of nicotinic receptor agonists and indirect trophic actions of nicotine on brain are suggested from observations describing nicotine as a cognitive enhancer by increasing vigilance and improving learning and memory. While an increasing number of studies have given evidence of neuroprotective and neurotrophic effects of nicotine treatment, the molecular mechanism mediating the neurotrophic effects of nicotine are not fully understood. Previously in an analysis of several neurotrophic factors as possible mediators of nicotine-induced neuroprotection and/or neurotrophic effects we could reveal that an acute intermittent nicotine treatment increases fibroblast growth factor-2 mRNA and protein in several brain regions of rat brain. Even if other studies have demonstrated in different paradigms that nicotine administration modulates expression level of a variety of genes, there is still a lack of indication which candidate genes, involved in neuroprotective responses are modulated by nicotine. In the present work we have used a microarray assay to further find and characterize new genes responsive to acute intermittent nicotine treatment and linked to neuroprotection. Therefore, we used Rat Genome U34A Affymetrix GeneChip arrays containing about 8800 probe sets to characterize transcriptional responses in the rat parietal cortex after acute intermittent nicotine treatment. We focused our attention to expression of transcription factors and several of them were up- or down-regulated by nicotine, among these Nr4a1 (Nurr77), Egr-1 and Egr-2. In situ hybridization was used to corroborate the microarray data and to reveal further spatial and temporal patterns of these nicotine induced genes. Taken together the present results identified several novel candidate genes modified by acute intermittent nicotine exposure and as such potentially involved in neuroprotective-neurotrophic actions. PMID- 15890457 TI - Effects of creatine treatment on the survival of dopaminergic neurons in cultured fetal ventral mesencephalic tissue. AB - Parkinson's disease is a disabling neurodegenerative disorder of unknown etiology characterized by a predominant and progressive loss of dopaminergic neurons in the substantia nigra. Recent findings suggest that impaired energy metabolism plays an important role in the pathogenesis of this disorder. The endogenously occurring guanidino compound creatine is a substrate for mitochondrial and cytosolic creatine kinases. Creatine supplementation improves the function of the creatine kinase/phosphocreatine system by increasing cellular creatine and phosphocreatine levels and the rate of ATP resynthesis. In addition, mitochondrial creatine kinase together with high cytoplasmic creatine levels inhibit mitochondrial permeability transition, a major step in early apoptosis. In the present study, we analyzed the effects of externally added creatine on the survival and morphology of dopaminergic neurons and also addressed its neuroprotective properties in primary cultures of E14 rat ventral mesencephalon. Chronic administration of creatine [5 mM] for 7 days significantly increased survival (by 1.32-fold) and soma size (by 1.12-fold) of dopaminergic neurons, while having no effect on other investigated morphological parameters. Most importantly, concurrent creatine exerted significant neuroprotection for dopaminergic neurons against neurotoxic insults induced by serum and glucose deprivation (P < 0.01), 1-methyl-4-phenyl pyridinium ion (MPP+) [15 microM] and 6 hydroxydopamine (6-OHDA) [90 microM] exposure (P < 0.01). In addition, creatine treatment significantly protected dopaminergic cells facing MPP+-induced deterioration of neuronal morphology including overall process length/neuron (by 60%), number of branching points/neuron (by 80%) and area of influence per individual neuron (by 60%). Less pronounced effects on overall process length/neuron and number of branching points/neuron were also found after 6-OHDA exposure (P < 0.05) and serum/glucose deprivation (P < 0.05). In conclusion, our findings identify creatine as a rather potent natural survival- and neuroprotective factor for developing nigral dopaminergic neurons, which is of relevance for therapeutic approaches in Parkinson's disease and for the improvement of cell replacement strategies. PMID- 15890458 TI - Responses to positive and negative smoking-related images: effects of current smoking status and degree of smoking addiction. AB - The literature indicates that stimulant users of various dependency levels endorse and react differently to stimuli that portray the stimulant either positively or negatively, however these studies have not been extended to smoking. Here pictures are used to depict either positive or negative connotations of smoking. The current study concentrated on smokers with different levels of dependency. Seventy-three digital images were rated on both a smoking and emotional content scale. The pictures were rank ordered, yielding the 10 most positive and negative smoking-related pictures. Emotional content scores for these pictures were also recorded. Data from 148 subjects [light- (n=28), heavy- (n=17), ex- (n=32) and never-smokers (n=71)] were analysed. Using a mixed factorial ANOVA light- and heavy-smokers were found to score positive pictures significantly more positive than the never-smokers, they did not vary on negative or emotional ratings. Thus positive pictures did not distinguish heavy and light levels of substance use as suggested by the literature and this was not influenced by emotional content. PMID- 15890459 TI - Preliminary study on inhibition of genotoxicity by piperine in mice. AB - A significant suppression (33.9-66.5%) in the micronuclei formation induced by benzo(a)pyrene and cyclophosphamide was reduced following oral administration of piperine at doses of 25, 50 and 75 mg/kg in mice. PMID- 15890460 TI - Photoprotective activity of Buddleja scordioides. AB - The purpose of this study was to determine the photoprotective properties of the methanolic extract of Buddleja scordioides, as well as verbascoside and linarin which were isolated from this extract, and linarin acetate prepared in the laboratory. The photoprotective effect of substances against UV-B induced cellular death was evaluated by challenge experiments using Escherichia coli. Verbascoside and linarin acetate showed the highest protection. The sun protection factor (SPF) of the methanolic extract, linarin, linarin acetate, and verbascoside was evaluated by guinea pig bioassays. Verbascoside showed the largest SPF measurement. PMID- 15890461 TI - Cytotoxic activity of embelin from Lysimachia punctata. AB - A major benzoquinone pigment, embelin (1), was isolated from the underground parts of Lysimachia punctata. Compound 1 showed a significant cytotoxic activity in vitro against B16 and XC cell lines with ED50 values of 13 microg/ml and 8 microg/ml, respectively. PMID- 15890462 TI - Antimicrobial and cytotoxic activity of Ruta graveolens. AB - The methanol, petroleum ether, ethyl acetate and water-methanol extracts of Ruta graveolens were found to possess antimicrobial and cytotoxic activities. PMID- 15890463 TI - Analgesic and anti-inflammatory activity of Leonurus sibiricus. AB - The methanolic extract of Leonurus sibiricus aerial parts injected intraperitoneally at dose of 250 and 500 mg/kg showed a significant analgesic effect in acetic acid-induced writhing in mice. Moreover, when given orally to rats at dose of 200 and 400 mg/kg, it showed a significant anti-inflammatory activity against carrageenin induced rat paw edema in rats. PMID- 15890464 TI - Quantitative and cytotoxic activity determinations on Galanthus nivalis subsp. cilicicus. AB - Aerial and underground parts of Galanthus nivalis subsp. cilicicus, a wild growing species in Turkey, were collected during two different vegetation periods in flowering and fruiting seasons. Herba and bulbus Galanthi were prepared from each specimen. With the aim of collecting data for prospective monographs on this drug, contents of humidity, ash, sulphated ash and total alkaloids were determined according to DAB 10. The specimens were also analyzed quantitatively for two of the principal alkaloids of the genus, galanthamine and lycorine, by using a method based on spectrophotometry complemented with TLC. LC50 values were determined for the ethanolic and alkaloidal extracts of each of the specimens using brine shrimp lethality bioassay. PMID- 15890465 TI - Antimicrobial activity of clerodane diterpenoids from Polyalthia longifolia seeds. AB - The diterpenoids 16alpha-hydroxy-cleroda-3,13 (14)-Z-diene-15,16-olide (1) and 16 oxo-cleroda-3, 13(14)-E-diene-15-oic acid (2), isolated from the hexane extract of the seeds of Polyalthia longifolia, demonstrated significant antibacterial and antifungal activities. PMID- 15890466 TI - Preliminary assay on the radical scavenging activity of olive wood extracts. AB - The dichloromethane and ethanol extracts of Olea europaea wood (picual olive cultivar) were screened for antioxidant activity, determined by the DPPH free radical scavenging assay. The ethanol extract displayed potent antioxidant activity. PMID- 15890467 TI - Antileishmanial and antifungal activities of xanthanolides isolated from Xanthium macrocarpum. AB - Seven xanthanolides, xanthinosin, xanthatin, 4-hydroxyxanthinosin, xanthinin, 4 epiisoxanthanol, 4-epixanthanol, 2-hydroxyxanthinosin and 4-oxobedfordia acid, were isolated from the fruits of Xanthium macrocarpum. A valuation of the antifungal activity of these xanthanolides against Candida albicans, Candida glabrata and Aspergillus fumigatus and of their antileishmanial activity against Leishmania infantum and Leishmania mexicana is presented. PMID- 15890468 TI - Antimicrobial activity of ten Lycoperdaceae. AB - The 60% methanolic extracts of ten Lycoperdaceae exhibited antimicrobial activity. PMID- 15890469 TI - Quenching of singlet molecular oxygen by Commiphora myrrha extracts and menthofuran. AB - The quenching activity against singlet oxygen, an actor of lipid peroxidation and DNA degradation, of the essential oil and resinoid of Commiphora myrrha from Somalia has been studied and compared to DL-alpha-tocopherol using 1,3 diphenylisobenzofuran (DPBF) as a probe. To insure that the furan ring was the site of the reaction, experiments were conducted with menthofuran. The essential oil and menthofuran show a higher activity than DL-alpha-tocopherol, suggesting their potential usefulness to neutralise this deleterious form of molecular oxygen. PMID- 15890470 TI - Traditional phytotherapy of the Albanians of Lepushe, Northern Albanian Alps. AB - An ethnobotanical and ethnopharmacognostic survey has been carried out in one of the most isolated mountainous area in Europe: the village of Lepushe and its surrounding territory, in the Northern Albanian Alps. Approximately 70 botanical taxa and 160 preparations, mainly derived from plants, but also derived from animal products or minerals, have been recorded. The archaic belief of the signature still plays a very important role in the present ethnomedicine of the Albanians of Lepushe. As a consequence, aerial parts of Chelidonium majus are used to treat jaundice; leaves of the fern Phyllitis scolopendrium are thought to be able to treat every respiratory and lung affection; the bulbs of Lilium martagon are used to treat liver diseases; jasper is rubbed into milk and given to sheep to drink and, the membrane of a hen's muscular stomach is used to treat human kidney stones. A current common ethnomedical use of P. scolopendrium and L. martagon is recorded for the first time in Europe. PMID- 15890471 TI - Cytotoxicity of plants used in traditional medicine in Yemen. AB - Twenty-five extracts obtained from 14 plant species used in the traditional medicine in Yemen have been screened for cytotoxic activity against human ECV-304 cells. Extracts of Dracaena cinnabari, Eucalyptus camaldulensis, Euclea divinorum, Euphorbia cactus, Pulicaria crispa, and Withania somnifera displayed a remarkable activity. PMID- 15890472 TI - Antiviral evaluation of plants from Brazilian Atlantic Tropical Forest. AB - The antiviral activity of six medicinal plants from Brazilian Atlantic Tropical Forest was investigated against two viruses: herpes simplex virus type 1 (HSV-1) and poliovirus type 2 (PV-2). Cuphea carthagenensis and Tillandsia usneoides extracts showed the best antiherpes activity. T. usneoides dichloromethane, ethyl acetate and n-butanol extracts, and Lippia alba n-butanol extract showed inhibition of HSV-1, strain 29R/acyclovir resistant. In addition, only L. alba ethyl acetate extract showed antipoliovirus activity. These results corroborate that medicinal plants can be a rich source of potential antiviral compounds. PMID- 15890473 TI - Analgesic activity of Amorphophallus campanulatus tuber. AB - The methanol extract of Amorphophallus campanulatus tuber, given orally at the doses of 250 and 500 mg/kg, showed significant analgesic activity in mice. PMID- 15890474 TI - Sp1 and AP2 enhance promoter activity of the mouse GM3-synthase gene. AB - Promoters of the glycosyltransferase genes for ganglioside synthesis are TATA less and often have multiple binding sites for transcription factors Sp1 and AP2 in their proximal regions. However, the function of Sp1 and AP2 in the promoters has not yet been defined. Here, we cloned 5'-flanking fragments of the mouse GM3 synthase gene and assessed the promoter activity of these fragments in mouse Neuro-2a cells. This promoter is TATA-less and contains a number of potential transcription factor-binding sites. Multiple putative transcriptional initiation sites for this gene were identified, including several downstream initiation sites. We then set out to dissect the regulatory elements important for GM3 synthase promoter function. We found that a 5'-flanking 254-bp DNA fragment of the gene contained regulatory elements including two Sp1-binding and six AP2 binding sites that were essential for the basal activity of the promoter in mouse Neuro-2a cells. The effects of the individual Sp1- and AP2-binding sites on basal activity of the GM3-synthase gene were investigated. Mutations in the juxtaposed Sp1/AP2-binding site and in an AP2-binding site decreased the activity of the proximal promoter to approximately 50%. In vitro and in vivo interactions between transcription factors Sp1 and AP2 and these regulatory elements were confirmed by EMSA and the chromatin immunoprecipitation approach, respectively. Therefore, our results demonstrate that Sp1 and AP2 enhance the basal activity of the TATA-less mouse GM3-synthase promoter. PMID- 15890475 TI - The retrograde response links metabolism with stress responses, chromatin dependent gene activation, and genome stability in yeast aging. AB - Yeast can be used as a model to understand the impact mitochondria have on aging in higher organisms. Mitochondrial dysfunction increases with replicative age in yeast, and this is associated with the induction of the retrograde response. This intracellular signaling pathway from the mitochondrion to the nucleus results in changes in the expression of metabolic and stress genes, which adapt the yeast cell to the loss of tricarboxylic acid cycle activity by providing alternate anaplerotic sources of biosynthetic precursors. The induction of the retrograde response increases longevity. Paradoxically, it also leads to the production of extrachromosomal ribosomal DNA circles, which cause yeast demise. The deleterious effects of these circles are mitigated by the retrograde response, which increases longevity in part due to this effect and partly due to other activities. Rtg2p is the retrograde signal transducer proximal to the mitochondrion, and it interacts with several proteins in relaying the retrograde signal to the transcription factor Rtg1p-Rtg3p. Rtg2p also suppresses ribosomal DNA circle production. When it is engaged in retrograde signaling, it cannot fulfill the latter role. The SAGA-like SLIK complex is one of the protein complexes in which Rtg2p has been found. This histone acetyltransferase, transcriptional co-activator complex contains Gcn5p, and it potentiates the activation of retrograde responsive genes. SLIK complex integrity, and in particular Gcn5p, are needed for retrograde response extension of life span. Thus, the retrograde response through SLIK links metabolism, stress responses, chromatin-dependent gene regulation, and genome stability in yeast aging. Gene regulatory phenomena akin to the retrograde response also operate in human cells, which display both common and cell-type specific changes in gene expression on loss of mitochondrial function. PMID- 15890476 TI - Photogenotoxicity of hypericin in HaCaT keratinocytes: implications for St. John's Wort supplements and high dose UVA-1 therapy. AB - Extract of St. John's Wort (Hypericum perforatum) is commonly used as natural remedy for treatment of mild to moderate depression. However, it contains a powerful photoactive component, hypericin, which can cause a severe photodermatitis when eaten by grazing animals (hypericism). In humans, there is evidence that supplementation with St. John's Wort can reduce the minimal erythemal dose (MED) in patients undergoing high dose UVA-1 phototherapy. This is a recent development in phototherapy where the most erythemogenic parts of the UVA spectrum are filtered out, allowing delivery of higher doses of the longer wavelengths of UVA. Although current published evidence suggests that the plasma levels of hypericin are unlikely to cause clinical phototoxicity, it has been established that photoactive compounds can cause DNA damage at sub-toxic and sub erythemal doses, the effects of which might not be apparent for many years after the event. The present study used HaCaT keratinocytes to investigate the photoclastogenic ability of hypericin on irradiation with UVA. The results show that although the combination of hypericin and UVA light increased the genotoxic burden, when all factors are taken into account, the risk of significant photogenotoxic damage incurred by the combination of Hypericum extracts and UVA phototherapy may be low in the majority of individuals. PMID- 15890477 TI - Induction of phase-1 metabolizing enzymes by oltipraz, flavone and indole-3 carbinol enhance the formation and transport of benzo[a]pyrene sulfate conjugates in intestinal Caco-2 cells. AB - The small intestine is well equipped with various phase-1 and phase-2 xenobiotic metabolizing enzymes (XME), which contribute to the detoxification process of the body. Many XME are regulated via aryl hydrocarbon receptor (AhR)-dependent pathways, and numerous naturally occurring AhR agonists (e.g. flavonoids, dietary indoles) have been identified to date. In the present study we show that pretreatment of Caco-2 cells with food-associated compounds (flavone and indole-3 carbinol) and with the anticancer chemopreventive agent oltipraz enhances the formation of the major metabolites of the procarcinogen benzo[a]pyrene (BP) formed by intestinal Caco-2 cells, namely BP-1-sulfate and BP-3-sulfate, and their transport to the apical compartment of a Transwell chamber. Oltipraz treatment was most effective in this regard followed by flavone and indole-3 carbinol. The effect observed here after pretreatment with oltipraz, flavone and I3C was the result of the induction of both CYP1A1 and CYP1B1, as was confirmed by analysis of CYP1A1 (protein and mRNA) and CYP1B1 (mRNA) expression. In summary, our study shows that the induction of both CYP1A1 and CYP1B1 resulted in an accelerated metabolism and an enhanced clearance of the potent procarcinogen BP, indicating that flavone, indole-3-carbinol and oltipraz have an impact on the biochemical barrier against BP in intestinal cells. PMID- 15890478 TI - Under the skin: Biotransformation of para-aminophenol and para-phenylenediamine in reconstructed human epidermis and human hepatocytes. AB - We investigated the biotransformation of the oxidative arylamine (AA) hair dye ingredients [14C]-para-aminophenol (PAP) and [14C]-para-phenylenediamine (PPD) in reconstructed human epidermis and human hepatocytes. Human epidermis quantitatively transformed PAP to its N-acetylated derivative (APAP), whereas hepatocytes transformed PAP to sulfate or glucuronic acid conjugates of APAP or PAP as well as free APAP. Epidermis and hepatocytes converted PPD to N-mono- (MAPPD) and N,N'-di-acetylated (DAPPD) derivatives. At higher concentrations of PPD (250-1000 microM), epidermis or hepatocytes produced more of the MAPPD, whereas concentrations below 250 microM and lower favoured formation of the DAPPD metabolite. When compared with epidermis, human hepatocytes had a three-fold or eight-fold greater capacity for generation of MAPPD or DAPPD, respectively. No evidence of transformation of PAP or PPD to N-hydroxylated derivatives was found in epidermis or hepatocytes. Our results suggest that (i) after dermal absorption of PAP or PPD, humans are systemically exposed to acetylated derivatives; (ii) current in vitro skin absorption studies may be inadapated for determination of human systemic exposure to AAs due to reduced or absent metabolic capacity of non viable skin; (iii) due to qualitative differences between dermal and hepatic metabolism, oral toxicity studies may be unsuited for the hazard assessment of dermal exposure to AAs; and (iv) use of induced rodent liver S9 metabolic activation systems for in vitro genotoxicity studies may produce misleading results on the hazard of human dermal exposure to AAs. In conclusion, our data support the growing evidence that AAs are transformed in human skin and suggest that current practices of safety assessment of AAs should take these findings into account. PMID- 15890479 TI - Effects of enzyme inducers and inhibitors on the pharmacokinetics of intravenous torasemide in rats. AB - In order to find whether torasemide is metabolized via CYP isozymes in rats, torasemide at a dose of 2mg/kg was infused in rats pretreated with SKF 525-A, a non-specific CYP isozyme inhibitor in male Sprague-Dawley rats. The total area under the plasma concentration-time curve from time zero to time infinity (AUC) of torasemide was significantly greater in rats pretreated with SKF 525-A (a non specific CYP isozyme inhibitor in rats) than that in control rats (3570 versus 1350 microg min/ml). This indicated that torasemide is metabolized via CYP isozymes in rats. Hence, torasemide was infused in rats pretreated with various enzyme inducers and inhibitors to find what types of CYP isozymes are involved in the metabolism of torasemide in rats. The AUC values were not significantly different in rats pretreated with 3-methylcholanthrene, phenobarbital, isoniazid, quinine and troleandomycin (main inducers of CYP1A1/2, CYP2B1/2, and CYP2E1, and main inhibitors of CYP2D1 and CYP3A1/2 in rats, respectively) compared with those in respective control rats. However, in rats pretreated with dexamethasone (a main inducer of CYP3A1/2 in rats), the AUC was significantly smaller than that in control rats (1290 versus 1590 microg min/ml). Dexamethasone probably also induces rat CYP2C11; this could be due to an increase in CYP2C11 in rats pretreated with dexamethasone. It has been reported from our laboratories that in rats pretreated with sulfaphenazole (a main inhibitor of CYP2C11 in rats) the AUC was significantly greater than that in control rats (2970 versus 1610 microg min/ml). The above data suggested that torasemide could be metabolized in male rats mainly via CYP2C11. PMID- 15890480 TI - Effects of alkaloids of Himatanthus lancifolius (Muell. Arg.) Woodson, Apocynaceae, on smooth muscle responsiveness. AB - Himatanthus lancifolius, popularly known as "agoniada" in Brazil, is largely used in folk medicine against asthma, dysmenorrhea and as an emenagogue and abortive. This study reveals the effects of an alkaloid rich fraction (AlkF) obtained from the bark of Himatanthus lancifolius in vascular and non-vascular smooth muscle responsiveness. Incubation of AlkF (3-30 microg/ml) during 15 min generates a concentration-related and fully reversible reduction in maximal contractile responses evoked by acetylcholine and phenylephrine in rat jejune and aorta preparations, respectively. Exposition of endothelium-denuded pre-contracted rat aorta rings to AlkF results in a complete relaxation, with EC(50) of 22.2 (16.2 28.2 microg/ml). AlkF is also able to induce a concentration-related rightward shift of cumulative concentration curves for calcium in uterus and aorta rings maintained in depolarizing nutritive solution. Moreover, addition of AlkF in calcium-free solution also reduces, in a concentration-dependent manner, the ability of caffeine and phenylephrine to contract aorta rings. This study reveals that the bark of Himatanthus lancifolius possesses one or more indole alkaloids able to alter non-vascular and vascular smooth muscle responsiveness, an event that may involve the blocking of calcium entry or changes on intracellular calcium utilization or mobilization. PMID- 15890481 TI - Monoamine oxidase B (MAO-B) inhibition by active principles from Uncaria rhynchophylla. AB - Attenuation of monoamine oxidase B (MAO-B) activity may provide protection against oxidative neurodegeneration. For this reason, inhibition of MAO-B activity is used as part of the treatment of Parkinson's and Alzheimer's patients. The hook of Uncaria rhynchophylla (Miq.) Jacks. (Rubiaceae) is a traditional Chinese herbal drug that is generally used to treat convulsive disorders. In this study, the fractionation and purification of Uncaria rhynchophylla extracts using a bioguided assay isolated two known compounds, (+) catechin and (-)-epicatechin. The compounds inhibited MAO-B, as measured by an assay of rat brain MAO-B separated by electrophoresis on a 7.5% native polyacrylamide gel. The IC(50) values of (+)-catechin and (-)-epicatechin were 88.6 and 58.9 microM, respectively, and inhibition occurred in a dose-dependent manner, as measured by the fluorescence method. The Lineweaver-Burk plot revealed K(i) values for (+)-catechin and (-)-epicatechin of 74 and 21 microM, respectively. This suggests that these two compounds, isolated here for the first time from Uncaria rhynchophylla, might be able to protect against neurodegeneration in vitro, and, therefore, the molecular mechanism deserves further study. This finding may also increase interest in the health benefits of Uncaria rhynchophylla. PMID- 15890482 TI - Preoperative hepatic 3D models: virtual liver resection using three-dimensional imaging technique. AB - Emerging new techniques for liver resections set new requirements for the preoperative imaging and planning. Open surgery is a three-dimensional procedure and planning of the resection line may be difficult when basing on conventional two-dimensional CTs or MRIs, although all the information is there. With multidetector-row CT (MDCT), thin slices can be obtained with excellent temporal resolution, and precise three-dimensional (3D) models can be created. We regard 3D imaging technique useful in most liver resections. It improves the surgeon's knowledge of liver anatomy and makes even more complicated liver resections safe. Better knowledge of three-dimensional appearances of liver structures may further improve the results of curative liver surgery. However, before becoming a routine clinical procedure, research and development are still needed. Also, careful testing and evaluation of the methods have to be performed. In the future, 3D models will probably play an important role in the preoperative planning of liver resections. PMID- 15890483 TI - Use of prior mammograms in the classification of benign and malignant masses. AB - The purpose of this study was to determine the importance of using prior mammograms for classification of benign and malignant masses. Five radiologists and one resident classified mass lesions in 198 mammograms obtained from a population-based screening program. Cases were interpreted twice, once without and once with comparison of previous mammograms, in a sequential reading order using soft copy image display. The radiologists' performances in classifying benign and malignant masses without and with previous mammograms were evaluated with receiver operating characteristic (ROC) analysis. The statistical significance of the difference in performances was calculated using analysis of variance. The use of prior mammograms improved the classification performance of all participants in the study. The mean area under the ROC curve of the readers increased from 0.763 to 0.796. This difference in performance was statistically significant (P = 0.008). PMID- 15890484 TI - Progress toward automated metabolic profiling of human serum: comparison of CPMG and gradient-filtered NMR analytical methods. AB - The investigation of drug delivery and metabolism requires the analysis of molecules in complicated biological matrices such as human serum. In NMR-based metabonomic analysis, T(2) relaxation editing with a CPMG filter is commonly used to suppress background signals from proteins and other endogenous components. Radio frequency pulse imperfections and incomplete irradiation across the spectral bandwidth can cause phase and baseline distortions in CPMG spectra. These distortions are exacerbated by water suppression techniques. Baseline correction methods included in commercially available data processing software packages may be incapable of producing artifact-free spectra. To increase the analytical precision of metabolic profiling, one NMR spectroscopist may be responsible for manually phasing and baseline correcting hundreds of spectra individually to remove operator-dependent variations, significantly reducing throughput. For metabonomic analysis of human serum, it was observed that the application of a pulsed field gradient filter produced (1)H NMR spectra well suited to automatic phasing routines. Superior baseline characteristics, an increased tolerance to radio frequency pulse imperfections, and improved water suppression were achieved. A concomitant reduction in signal intensity compared with the CPMG method was easily recovered by increasing the number of scans. Principal component analysis (PCA) of spectra, acquired under a variety of experimental conditions, revealed the improved reproducibility and robustness of (1)H NMR pulsed field gradient-filtered metabonomic analyses of serum compared to the CPMG method. PMID- 15890485 TI - Prediction of drug absorption based on immobilized artificial membrane (IAM) chromatography separation and calculated molecular descriptors. AB - The aim of this study was to evaluate the usefulness of IAM chromatography in building a model that would allow prediction of drug absorption in humans. The human intestinal absorption values (%HIA) for 52 drugs with low to high intestinal absorption were collected from the literature. The retention (capacity factor, k') of each drug was measured by reverse-phase HPLC using an IAM.PC.DD2 column (prepared with phosphatidylcholine analogs, 12 microM, 300A, 15 cm x 4.6 mm) with an eluent of acetonitrile-0.1M phosphate buffer at pH 5.4. In addition, 76 molecular descriptors and solubility parameters for each drug were calculated using ChemSW from the 3D-molecular structures. Stepwise regression was employed to develop a regression equation that would correlate %HIA with molecular descriptors and k'. Human intestinal absorption was reciprocally correlated to the negative value of the capacity factor (-1/k') (R=0.64). The correlation was further improved with the addition of molecular descriptors representing molecular size and shape (molecular width, length and depth) solubility (solubility parameter, HLB, hydrophilic surface area) and polarity (dipole, polar surface area) (R=0.83). Experimentally measured IAM chromatography retention values and calculated molecular descriptors and solubility parameters can be used to predict intestinal absorption of drugs in humans. Developed QSAR can be used as a screening method in the designing of drugs with appropriate IA and for the selection of drug candidates in the early stage of drug discovery process. PMID- 15890486 TI - Femur window--a new approach to microcirculation of living bone in situ. AB - BACKGROUND: The processes of osteogenesis, bone remodelling, fracture repair and metastasis to bone are determined by complex sequential interactions involving cellular and microcirculatory parameters. Consequently studies targeting the analysis of microcirculatory parameters on such processes should mostly respect these complex conditions. However these conditions could not yet be achieved in vitro and therefore techniques that allow a long-term observation of functional and structural parameters of microcirculation in bone in vivo at a high spatial resolution are needed to monitor dynamic events, such as fracture healing, bone remodelling and tumor metastasis. METHODS: We developed a bone chamber implant (femur window) for long-term intravital microscopy of pre-existing bone and its microcirculation at an orthotopic site in mice preserving the mechanical properties of bone. After bone chamber implantation vascular density, vessel diameter, vessel perfusion, vascular permeability and leukocyte-endothelial interactions (LEIs) in femoral bone tissue of c57-black mice (n=11) were measured quantitatively over 12 days using intravital fluorescence microscopy. Furthermore a model for bone defect healing and bone metastasis in the femur window was tested. RESULTS: Microvascular permeability and LEIs showed initially high values after chamber implantation followed by a significant decrease to a steady state at day 6 and 12, whereas structural parameters remained unaltered. Bone defect healing and tumor growth was observed over 12 and 90 days respectively. CONCLUSION: The new femur window design allows a long-term analysis of structural and functional properties of bone and its microcirculation quantitatively at a high spatial resolution. Altered functional parameters of microcirculation after surgical procedures and their time dependent return to a steady state underline the necessity of long-term observations to achieve unaltered microcirculatory parameters. Dissection of the complex interactions between bone and microcirculation enables us to evaluate physiological and pathological processes of bone and may give new insights especially in dynamic events e.g. fracture healing, bone remodeling and tumor metastasis. PMID- 15890487 TI - Uncemented short-length diaphyseal segmental replacement prosthesis fixation- finite element analysis and long-term results. AB - INTRODUCTION: Extensively porous coated segmental replacement prostheses with intramedullary cementless fixation to bone over the whole length of stem often exhibit resorption of the surrounding bone due to stress-shielding. This makes them particularly susceptible to aseptic loosening. STUDY: A finite element analysis of the state of loading of a short-length fixation in a new prosthetic stem design has shown a definite advantage over long-length fixation. The stress pattern within the bone surrounding the prosthesis confirmed that shortening of the ongrowth area in length increases the stress values at the resection level significantly. This stem (Endlock) has been used for diaphyseal anchorage in the treatment of tumors in combination with an artificial joint of proven design in order to reduce stress shielding. RESULTS: No Endlock stem fractures or aseptic loosenings were observed at recent follow-up. The early clinical results comply with the theoretical assumptions. CONCLUSIONS: A short-length fixation system based on intramedullary anchorage of segmental replacement endoprostheses would possibly support physiologic adaptive processes more than fixation over the full length of the stem. PMID- 15890488 TI - Thyroid hormone availability and activity in avian species: a review. AB - The intracellular thyroid hormone (TH) availability is influenced by different metabolic pathways. Some of the changes in intracellular TH availability can be linked to changes in local deiodination and sulfation capacities. The secretion of the chicken thyroid consists predominantly of thyroxine (T4). TH receptors (TRs) preferentially bind 3,5,3'-triiodothyronine (T3). Therefore, the metabolism of T4 secreted by the thyroid gland in peripheral tissues, resulting in the production and degradation of receptor-active T3, plays a major role in thyroid function. Food restriction in growing chickens increases hepatic type III deiodinase (D3) levels but decreases growth hormone (GH)-dependent variables such as plasma insulin-like growth factor-I (IGF-I) and T3 concentrations. Refeeding restores hepatic D3 and plasma T3 to control levels within a few hours. It can be concluded that the tissue and time dependent regulation of the balance between TH activating and inactivating enzymes plays an essential role in the control of local T3 availability and hence in TH activity. Two separate genes encode multiple TR isoforms, i.e. TRalpha and TRbeta. These TRs consist of a DNA-binding domain, a ligand-binding domain, a hinge region and an amino-terminal (A/B) domain. TRs mediate their effects on transcription by binding as homodimers or heterodimers to the TH response elements (TREs). Also, unliganded TRs can bind to TREs and may so modulate transcription of target genes. PMID- 15890489 TI - Haplotype XV of the Y-chromosome is the main haplotype in West-Europe. AB - We have analyzed Y-chromosome variation in a large sample of males from Western Europe by surveying p49a,f TaqI polymorphisms. Haplotype XV (A3, Cl, D2, Fl, Il) is the main Y-chromosome haplotype in West Europe, with a Basque focus in Southwestern Europe. This study demonstrates that the geographic distribution of Y-chromosome variation for p49a,f TaqI haplotype XV reveals an important genetic identity for populations that live in the Occidental part of Europe. PMID- 15890490 TI - Differential effect of Cyclosporin A and FK506 on SPARC mRNA expression by human gingival fibroblasts. AB - BACKGROUND: Secreted protein acidic and rich in cysteine (SPARC) is a glycoprotein that mediates cell-matrix interactions. In adults, its expression is mostly limited to tissue undergoing remodeling. During the development of Cyclosporin A (CsA)-induced gingival overgrowth (GO) a remodeling of the connective compartment occurs. By contrast, clinical trials showed that FK506 is not related to GO. SPARC expression and its involvement in GO is unknown. Our aim was, therefore, to analyze the effect of CsA and FK506 on SPARC gene expression. METHODS: Cultured human gingival fibroblasts were incubated with CsA, FK506 or with their vehicle (VH) for 24, 48 and 72 h. SPARC gene expression was determined by RT-PCR. RESULTS: SPARC mRNA levels tended to increase 72 h after CsA treatment, whilst they are undetectable in FK506-treated fibroblasts, compared to VH. CONCLUSION: This gene expression profile is consistent with the involvement of SPARC in the mechanisms leading to the development of CsA-induced GO. By contrast, the undetectable SPARC mRNA levels in FK506-treated fibroblasts suggest that FK506 may be associated with a role of ECM stabilization, that does not induce GO. PMID- 15890491 TI - N-acetyl-beta-D-hexosaminidase from Trichomonas vaginalis: substrate specificity and activity of inhibitors. AB - Among chitinolytic activities previously described in Trichomonas vaginalis, N acetyl-beta-D-hexosaminidase (NAHase) was the enzyme system expressing the highest level of specific activity. We report here some biochemical characteristics of NAHase purified from T. vaginalis. We found at first that the use of 4-methylumbellifferyl-substrate was responsible for a substrate affinity for the enzyme, about 1000-fold higher than those when using p-nitrophenyl substrates (PNP). Whereas the optimum pH was 7.0 using PNP-substrate, it was at 4.5 using 4-methylumbelliferyl-substrate. Four different substrates were compared for their action on T. vaginalis NAHase and we have found that N-acetyl-beta-D glucosaminide substrate was the most specific. DTT had no effect on enzyme activity suggesting that thiol group are not involved at the catalytic site. The use of previously described inhibitors showed a positive correlation between trichomonacidal activity and NAHase inhibition. PMID- 15890492 TI - Effects of an oxidative stress on human hemoglobin: a multiwavelength visible spectrometry study. AB - This study was carried out to investigate hemoglobin behavior and the role of cell membrane during oxidative stress of human red blood cells induced by a water soluble radical initiator, 2,2'-azobis(amidino-propane) hydrochloride (AAPH) and compare the observed data to the one obtained with purified human haemoglobin solution. The different forms of hemoglobin were identified and quantified by multiwavelength visible spectrometry using multiple linear regression analysis. Hemolysis was quantified by the Drabkin method. Oxidative stress on purified hemoglobin solutions induced an early formation of Hb(+). In intact erythrocytes, no modified form of haemoglobin was found. Only the hemoglobin released by hemolysis in the extracellular medium was notified in the same way as purified haemoglobin. Thus, the cell membrane appears to protect intraerythrocytic hemoglobin toward an extracellular oxidative stress. Oxidative stress-induced by hemolysis does not seem to be due to changes in intraerythrocytic hemoglobin forms. PMID- 15890493 TI - TNF-alpha levels in cancer patients relate to social variables. AB - Tumor necrosis factor-alpha (TNF-alpha) is an important cytokine associated with tumor regression and increased survival time for cancer patients. Research evidence relates immune factors (e.g., natural killer (NK) cell counts, NK cell lysis, lymphocyte profile, and lymphocyte proliferation) to the frequency and quality of social relations among cancer patients. We hypothesized that disruptions in social relations would be associated with lower TNF-alpha responses, and conversely, that reports of positive changes in social relations correlate with stronger responses. A prospective design measured changes in social activity and relationship satisfaction with a partner in 44 breast cancer patients at the time of cancer diagnosis, and initial surgery and 12 months later. Results indicated that patients reporting increased social activities or satisfaction exhibited stronger stimulated TNF-alpha responses. This is the first study to link changes in patient social relations with a cancer-relevant immune variable. PMID- 15890494 TI - Cortical correlates of false expectations during pain intensity judgments--a possible manifestation of placebo/nocebo cognitions. AB - We investigated the effects of expectation on intensity ratings and somatosensory evoked magnetic fields and electrical potentials following painful infrared laser stimuli in six healthy subjects. The stimulus series contained trials preceded by different auditory cues which either contained valid, invalid or no information about the upcoming laser intensity. High and low intensities occurred equally probable across cue types. High intensity stimuli induced greater pain than low intensity across all cue types. Furthermore, laser intensity significantly interacted with cue validity: high intensity stimuli were perceived less painful and low intensity stimuli more painful following invalid compared to valid cues. The amplitude of the evoked magnetic field localized within the contralateral secondary somatosensory cortex (SII) at about 165 ms after laser stimuli varied also both with stimulus intensity and cue validity. The evoked electric potential peaked at about 300 ms after laser stimuli and yielded a single dipole source within a region encompassing the caudal anterior cingulate cortex and posterior cingulate cortex. Its amplitude also varied with stimulus intensity, but failed to show any cue validity effects. This result suggests a priming of early cortical nociceptive sensitivity by cues signaling pain severity. A possible contribution of the SII cortex to the manifestation of nocebo/placebo cognitions is discussed. PMID- 15890495 TI - Delayed response and lack of habituation in plasma interleukin-6 to acute mental stress in men. AB - Acute mental stress induces a significant increase in plasma interleukin (IL)-6 levels as a possible mechanism for how psychological stress might contribute to atherosclerosis. We investigated whether the IL-6 response would habituate in response to a repetitively applied mental stressor and whether cortisol reactivity would show a relationship with IL-6 reactivity. Study participants were 21 reasonably healthy men (mean age 46+/-7 years) who underwent the Trier Social Stress Test (combination of a 3-min preparation, 5-min speech, and 5-min mental arithmetic) three times with an interval of 1 week. Plasma IL-6 and free salivary cortisol were measured immediately before and after stress, and at 45 and 105 min of recovery from stress. Cortisol samples were also obtained 15 and 30 min after stress. Compared to non-stressed controls, IL-6 significantly increased between rest and 45 min post-stress (p=.022) and between rest and 105 min post-stress (p=.001). Peak cortisol (p=.034) and systolic blood pressure (p=.009) responses to stress both habituated between weeks one and three. No adaptation occurred in diastolic blood pressure, heart rate, and IL-6 responses to stress. The areas under the curve integrating the stress-induced changes in cortisol and IL-6 reactivity were negatively correlated at visit three (r=-.54, p=.011), but not at visit one. The IL-6 response to acute mental stress occurs delayed and shows no adaptation to repeated moderate mental stress. The hypothalamus-pituitary-adrenal axis may attenuate stress reactivity of IL-6. The lack of habituation in IL-6 responses to daily stress could subject at-risk individuals to higher atherosclerotic morbidity and mortality. PMID- 15890496 TI - Neuropsychological correlates of word identification in Down syndrome. AB - In order to better understand the neuropsychological underpinnings of the relative strength in word identification in individuals with Down syndrome, the performance of children and adolescents with Down syndrome (N=29) was compared to the performance of a nonverbal-IQ matched group of children and adolescents with developmental disabilities of mixed etiologies (N=20) on measures letter/word identification and cognitive-linguistic functioning. Though no between-group differences were observed for letter/word identification or visual processing performance, individuals with Down syndrome showed significantly poorer verbal short-term memory and receptive vocabulary skills. In terms of neuropsychological correlates of letter/word identification, significant linear associations were observed between letter/word identification (K-ABC reading/decoding) and verbal short-term memory (K-ABC number recall), as well as receptive vocabulary (PPVT III) and visual processing (MVPT-R) in both groups. However, when only children with word identification competence (as opposed to letter identification competence) were included in analyses, visual perception scores (total MVPT-R) were significantly associated with word identification in the Down syndrome group, but not in the mixed comparison group. Implications for etiology-specific instructional approaches are discussed. PMID- 15890497 TI - Cortical gene expression in the neonatal ventral-hippocampal lesion rat model. AB - Schizophrenia is a chronic, debilitating psychotic illness of unknown etiology that has been the subject of many genetic studies. We studied the neonatal ventral-hippocampal lesioned rat as an animal model of schizophrenia in order to identify novel candidate genes for schizophrenia. Temporal and frontal cortices were assessed using cDNA microarrays for differences in mRNA expression associated with the lesion, haloperidol treatment and in two rat strains with differential sensitivity to the behavioural effects of the lesion. Genes that had altered expression levels as a result of the lesion, that were normalized by haloperidol treatment, and that differed between rat strains were selected. The pattern of differential transcription was confirmed with quantitative PCR for all six candidate genes: large conductance calcium-activated potassium channel, subfamily M, beta member 1 (Kcnmb1); doublecortex (dcx); adenylyl cyclase associated protein 1 (CAP1); adenosine monophosphate deaminase 2-isoform L (AMPD2); malic enzyme 3, NADP(+)-dependent, mitochondrial (Me3); and aspartylglucosaminidase (AGA). None of these genes has been extensively studied in schizophrenia, and further work with post-mortem tissue and genetic studies are ongoing. PMID- 15890498 TI - Quinolone resistance in bacteria: emphasis on plasmid-mediated mechanisms. AB - Bacterial resistance to quinolones/fluoroquinolones has emerged rapidly and such resistance has traditionally been attributed to the chromosomally mediated mechanisms that alter the quinolone targets (i.e. DNA gyrase and topoisomerase IV) and/or overproduce multidrug resistance efflux pumps. However, the discovery of the plasmid-borne quinolone resistance determinant, named qnr, has substantially broadened our horizon on the molecular mechanisms of quinolone resistance. Several recent reports of Qnr or its homologues encoded by transferable plasmids in Gram-negative bacteria isolated worldwide highlight the significance of the emerging plasmid-mediated mechanism(s). This also alerts us to the potential rapid dissemination of quinolone resistance determinants. Qnr belongs to the pentapeptide repeat family and protects DNA gyrase from the action of quinolone agents including the newer fluoroquinolones. This protection interplays with chromosomal mechanisms to raise significantly the resistance levels. The qnr-bearing strains generate quinolone-resistant mutants at a much higher frequency than those qnr-free strains. Furthermore, the qnr-plasmids are integron-associated and carry multiple resistance determinants providing resistance to several classes of antimicrobials including beta-lactams and aminoglycosides. The high quinolone resistance rates in Escherichia coli are used to address issues of quinolone resistance, and possible strategies for minimising quinolone resistance are discussed. PMID- 15890499 TI - Microbiological spectrum and susceptibility patterns of pathogens causing bacteraemia in paediatric febrile neutropenic oncology patients: comparison between two consecutive time periods with use of different antibiotic treatment protocols. AB - This study was devised to look at trends in the microbiological spectrum and susceptibility patterns of pathogens causing bacteraemia in paediatric febrile oncology patients. The retrospective study compared various microbiological aspects recorded for febrile oncology neutropenic patients treated with two different empirical antibiotic regimens (ceftazidime plus gentamicin during 1998 1999 and piperacillin/tazobactam plus amikacin during 2000-2002). Eighty-one bacteraemic episodes occurred in 41 patients. Overall, 132 (34 during 1998-1999 and 98 during 2000-2002) organisms were isolated: 84 (65%) Gram-negative bacteria, 39 (30%) Gram-positive bacteria and 7 (5%) fungi. Enterobacter spp. incidence decreased from 18 to 6% (P=0.07) while the recovery rates of Gram positive organisms increased from 24 to 32% (P=0.4) during 2000-2002 compared with 1998-1999. MRSA were not isolated from any episode of bacteraemia. Five (18%) of the 28 Escherichia coli and Klebsiella spp. isolates were beta-lactamase producers (80% [4/5] isolated during 2000-2002). Twenty-seven of 28, 27/27, 23/28, 20/25 and 27/28 of these isolates were susceptible to imipenem, piperacillin/tazobactam, gentamicin, ceftazidime and ciprofloxacin, respectively. Thirty-two of 34 (94%) and 60/74 (81%) of the Gram-negative organisms isolated during 2000-2002 were susceptible to piperacillin/tazobactam and ceftazidime, respectively (P=0.076). No major differences in the microbial spectrum and antibiotic susceptibilities were recorded between the two consecutive study periods. An increase in the number of extended beta-lactamase producing E. coli and Klebsiella spp. occurred during 2000-2002. All beta-lactamase producing organisms were susceptible to piperacillin/tazobactam and initial empirical therapy with piperacillin/tazobactam was more appropriate than ceftazidime to cover most of the pathogens causing bacteraemia. PMID- 15890500 TI - Biochemical and genetic characterization of the beta-lactamases of Y. aldovae, Y. bercovieri, Y. frederiksenii and "Y. ruckeri" strains. AB - The beta-lactamases of five strains each of Y. aldovae and "Y. ruckeri", and 10 strains each of Y. bercovieri and Y. frederiksenii were examined phenotypically and genetically. Beta-lactamase activity and induction assays and SDS-PAGE were applied for phenotypic characterization of these enzymes. Genotypically, PCR experiments applying degenerated primer pairs for the detection of AmpC beta lactamase genes were performed. All yersiniae yielded specific amplification products for ampC and all these strains expressed beta-lactamases. Each species produced its own, species-specific AmpC beta-lactamase. Inducibility of these enzymes was shown for Y. bercovieri, but not for the low-level enzyme producing species Y. aldovae and "Y. ruckeri". In contrast to these species, induction tests for Y. frederiksenii revealed heterogeneous results. Whereas the beta lactamases of 6 of 10 strains were inducible, the enzyme activities after induction in the remaining four were similar to those measured without an inducer. In addition to the AmpC enzyme, all Y. frederiksenii strains expressed a second beta-lactamase belonging to Ambler class A. The present study enlarges the knowledge about the beta-lactamases of four novel Yersinia species that are likely to be involved in human disease. Beta-lactamases of Y. aldovae and "Y. ruckeri" have been characterized for the first time. PMID- 15890501 TI - Susceptibility to moxifloxacin of pneumococci isolated in English hospitals participating in the European Antimicrobial Resistance Surveillance System (EARSS) in 2003. AB - Susceptibility to moxifloxacin, penicillin and erythromycin was determined for 592 invasive Streptococcus pneumoniae collected from 20 English hospitals participating in the European Antimicrobial Resistance Surveillance System (EARSS) during 2003. Resistance to moxifloxacin, penicillin and erythromycin was observed in 0.8%, 5.4% and 13% of the isolates, respectively. These results show that the large majority of pneumococci were susceptible to moxifloxacin in 2003, the year when it was licensed for clinical use in the UK. PMID- 15890502 TI - Electrophysiological correlates of facial decision: insights from upright and upside-down Mooney-face perception. AB - We investigated the ERP correlates of the subjective perception of upright and upside-down ambiguous pictures as faces using two-tone Mooney stimuli in an explicit facial decision task (deciding whether a face is perceived or not in the display). The difficulty in perceiving upside-down Mooneys as faces was reflected by both lower rates of "Face" responses and delayed "Face" reaction times for upside-down relative to upright stimuli. The N170 was larger for the stimuli reported as "faces". It was also larger for the upright than the upside-down stimuli only when they were reported as faces. Furthermore, facial decision as well as stimulus orientation effects spread from 140-190 ms to 390-440 ms. The behavioural delay in 'Face' responses to upside-down stimuli was reflected in ERPs by later effect of facial decision for upside-down relative to upright Mooneys over occipito-temporal electrodes. Moreover, an orientation effect was observed only for the stimuli reported as faces; it yielded a marked hemispheric asymmetry, lasting from 140-190 ms to 390-440 ms post-stimulus onset in the left hemisphere and from 340-390 to 390-440 ms only in the right hemisphere. Taken together, the results supported a preferential involvement of the right hemisphere in the detection of faces, whatever their orientation. By contrast, the early orientation effect in the left hemisphere suggested that upside-down Mooney stimuli were processed as non face objects until facial decision was reached in this hemisphere. The present data show that face perception involves not only spatially but also temporally distributed activities in occipito temporal regions. PMID- 15890503 TI - Enhanced oral paclitaxel absorption with vitamin E-TPGS: effect on solubility and permeability in vitro, in situ and in vivo. AB - Solubility and permeability being important determinants of oral drug absorption, this study was aimed to investigate the effect of D-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS) on the solubility and intestinal permeability of paclitaxel in vitro, in situ and in vivo, in order to estimate the absorption enhancement ability of TPGS. Aqueous solubility of paclitaxel is significantly enhanced by TPGS, where a linear increase was demonstrated above a TPGS concentration of 0.1 mg/ml. Paclitaxel demonstrated asymmetric transport across rat ileum with significantly greater (26-fold) basolateral-to-apical (B-A) permeability than that in apical-to-basolateral (A-B) direction. Presence of P glycoprotein (P-gp) inhibitor, verapamil (200 microM), diminished asymmetric transport of paclitaxel suggesting the role of P-gp-mediated efflux. TPGS showed a concentration-dependent increase in A-B permeability and decreased B-A permeability. The maximum efflux inhibition activity was found at a minimum TPGS concentration of 0.1 mg/ml, however, further increase in TPGS concentration resulted in decreased A-B permeability with no change in B-A permeability. Thus, the maximum paclitaxel permeability attained with 0.1 mg/ml TPGS was attributed to the interplay between TPGS concentration dependent P-gp inhibition activity and miceller formation. In situ permeability studies in rats also demonstrated the role of efflux in limiting permeability of paclitaxel and inhibitory efficiency of TPGS. The plasma concentration of [14C]paclitaxel following oral administration (25 mg/kg) was significantly increased by coadministration of TPGS at a dose of 50 mg/kg in rats. Bioavailability is enhanced about 4.2- and 6.3 fold when [14C]paclitaxel was administrated with verapamil (25 mg/kg) and TPGS, respectively, as compared to [14C]paclitaxel administered alone. The effect of verapamil on oral bioavailability of [14C]paclitaxel was limited relative to the TPGS, consistent with the in vitro solubility and permeability enhancement ability of TPGS. In conclusion, the current data suggests that the coadministration of TPGS may improve the bioavailability of BCS class II-IV drugs with low solubility and/or less permeable as a result of significant P-gp mediated efflux. PMID- 15890504 TI - [Hereditary angioneurotic edema: a case report in a 3-year-old child]. AB - Hereditary angioneurotic edema is a dominant autosomal disease (incidence 1/150,000), whose diagnosis is crucial as this condition can lead to fatal asphyxia within minutes. We report the case of a three-year-old girl, misdiagnosed as allergic asthma. Recognition of the syndrome led to adapted care restoring normal family and school life. Pathogenic cascades involved and therapeutic principles of this disease are reviewed. PMID- 15890505 TI - [2 years of hearing screening of high-risk infants and children in Martinique]. PMID- 15890506 TI - Burn-induced oxidative injury of the gut is ameliorated by the leukotriene receptor blocker montelukast. AB - There is increasing evidence that oxidative stress has an important role in the development of multiorgan failure after major burn injury. In the present study, we investigated whether the leukotriene receptor blocker montelukast is protective against burn-induced injury of the gut. Under brief ether anaesthesia, shaved dorsum of the rats was exposed to 90 degrees C (burn group) or 25 degrees C (control group) water bath for 10 s. Montelukast (10 mg/kg) or saline was administered intraperitoneally immediately after and at the 12th hour of the burn injury. Rats were decapitated 24 h after burn injury and the skin samples, as well as tissue samples from stomach, ileum and colon, were taken for the determination of malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and collagen contents. Tissues were also examined microscopically. Tumor necrosis factor-alpha (TNF-alpha) and lactate dehydrogenase (LDH) were assayed in serum samples. Severe skin scald injury (30% of total body surface area) caused a significant decrease in GSH level, which was accompanied with significant increases in MDA level, MPO activity and collagen content of tissues. Similarly, serum TNF-alpha and LDH were elevated in the burn group as compared to control group. On the other hand, montelukast treatment reversed all these biochemical indices, as well as histopathological alterations, which were induced by thermal trauma. Findings of the present study suggest that montelukast possesses an anti-inflammatory effect on burn-induced gastrointestinal damage and protects against oxidative injury by a neutrophil dependent mechanism. PMID- 15890507 TI - Effect of chemotherapy on serum end-products of lipid peroxidation in patients with small cell lung cancer: association with treatment results. AB - Many anti-cancer drugs induce formation of lipid peroxidation products that are toxic for lung cancer cells in vitro. We tested whether changes of serum thiobarbituric acid reactive substances (TBARs) and Schiff's bases (SB) are associated with treatment efficacy in 37 small cell lung cancer (SCLC) patients. Subjects received carboplatin (350 mg/m2, i.v.-Day 1), vincristine (1.3 mg/m2, i.v.-Day 1), and etoposide (120 mg/m2, oral dose-Days 1-4). Then 5 subsequent cycles were repeated at 21-day intervals. Serum TBARs and SB were measured fluorimetrically before and 6, 24h after introduction of the 1st, 3rd and 6th cycles. TBARs and SB levels rose 24 h after 1st chemotherapy in the whole group (2.5+/-1.4 vs. 4.2+/-2.0 micromol/dl, P<0.001 and 26.3+/-16.7 vs. 29.7+/ 9.8U(430)/ml, P<0.01, respectively) and the highest increments were in 19 patients with complete or partial response after 1st, 3rd and 6th cycles. In 9 subjects with progressive disease occurring before the 2nd cycle (early progression) TBARs and SB decreased 6 and 24h after the 1st cycle (4.3+/-1.2 vs. 3.4+/-1.4, P<0.05 vs. 2.7+/-0.9 micromol/dl, P<0.05 and 50.2+/-17.0 vs. 36.7+/ 13.2, P<0.05 vs. 36.5+/-13.4 U(430)/ml, P<0.01, respectively). Patients survival correlated with the 1st cycle-induced TBARs (r=0.49, P<0.001) and SB (r=0.56, P<0.002) increments. Subjects with negative SB and TBARs increments (n=8) had shorter survival than those (n=29) with positive increments in lipid peroxidation products (log rank test P<0.005). Monitoring of circulatory TBARs and SB may be helpful for screening of SCLC patients with high risk of early disease progression and chemotherapy failure. PMID- 15890508 TI - Predictors of hospital outcome and intubation in COPD patients admitted to the respiratory ICU for acute hypercapnic respiratory failure. AB - BACKGROUND: Mortality rate, the possible factors affecting mortality and intubation in patients with acute exacerbation of chronic obstructive pulmonary diseases (COPD) and hypercapnic respiratory failure (RF) are yet unclear. OBJECTIVE: To identify the possible factors affecting mortality and intubation in COPD patients. DESIGN: A prospective study using data obtained over the first 24h of respiratory intensive care unit (RICU) admission. Consecutive admissions of 656 patients were monitored and 151 of them who had acute exacerbation of COPD and hypercapnic RF were enrolled. SETTING: University hospital, Department of Chest Diseases, RICU. RESULTS: Mean age was 65.1 years. The mean APACHE II score was 23.7. Eighty-seven patients (57.6%) received mechanical ventilation (MV) via an endotracheal tube for more than 24 h. Twenty-two patients received non invasive ventilation (NIV). Fifty patients died (33.1%) in hospital during the study period. The mortality rate was 52.9% in patients in need of MV. In the multivariate analysis, the need for intubation, inadequate metabolic compensation for respiratory acidosis, and low (=bad) Glasgow Coma Score (GCS) were determined as independent factors associated with mortality. The low GCS (OR: 0.61; CI: 0.48 0.78) and high APACHE II score (OR: 1.24; CI: 1.11-1.38) were determined as factors associated with intubation. CONCLUSION: The most important predictors related to hospital mortality were the need for invasive ventilation and complications to MV. Adequate metabolic compensation for respiratory acidosis at admittance is associated with better survival. A high APACHE II score and loss of consciousness (low GCS) were independent predictors of a need to intubate patients. PMID- 15890509 TI - Exacerbations as a starting point of pro-active chronic obstructive pulmonary disease management. AB - Chronic obstructive pulmonary disease (COPD) exacerbations could represent an opportunity for pro-active COPD management rather than mere treatment if previously unknown disease is discovered; the extent of underdiagnosis and undertreatment of COPD in patients attending an emergency department (ED) with an exacerbation is not known. During 2002, we recalled 131 COPD patients in stable conditions, 4-8 weeks after they had attended the ED or been discharged from our University Hospital (North-West of Italy). Information on diagnosis and management prior to the ED attendance were collected; spirometry and arterial blood gas analyses were performed. One-third of patients had never been diagnosed and treated even though 83% of them had moderate-to-very-severe COPD and about 30% already had respiratory failure. Only 20% had received information on the nature of the disease and none had received a written action plan. Only 60% were receiving long-acting bronchodilators and 41% of patients with respiratory failure were receiving long-term oxygen. A substantial number of undiagnosed and untreated patients with moderate-to-very-severe COPD came to our attention through an exacerbation. This enforces the importance of exacerbations as the starting point of pro-active COPD management and of the ED as a valuable sentinel to identify this subset of patients. PMID- 15890510 TI - Systemic inflammatory response to exhaustive exercise in patients with chronic obstructive pulmonary disease. AB - Systemic inflammation may be present in patients with chronic obstructive pulmonary disease (COPD). Exercise is known to elicit an inflammatory response. We hypothesized that the systemic inflammatory response to exercise might be exaggerated in COPD patients compared to healthy subjects. Sixteen COPD patients and 11 healthy subjects performed a maximal incremental bicycle test. Before and at maximal exercise arterial blood samples were taken to determine circulating catecholamines, (subsets of) leukocytes, acute phase proteins, creatine kinase and myoglobin. At rest, increased levels of norepinephrine and systemic inflammation were present in COPD. The response of catecholamines to exercise was lower in COPD patients (P<0.01), which in part was due to the lower maximal exercise capacity of these patients (P<0.01). Exercise-induced leukocytosis showed similar responses in both groups, but occurred at higher levels in COPD. Although patients had increased levels of CRP at rest (P<0.001), exercise did not affect acute phase proteins. No systemic signs of muscle damage were found. The present study shows that COPD patients are exposed to systemic inflammation that is intensified by exhaustive exercise. The inflammatory response in COPD is not exaggerated compared to healthy subjects but occurs at a higher level and is observed at lower external workload. PMID- 15890511 TI - New insights in allergen avoidance measures for mite and pet sensitized patients. A critical appraisal. AB - It is widely acknowledged that avoidance of allergens such as those derived from foods, drugs, latex and stinging insects results in a complete disappearance of symptoms. By contrast, although it has been clearly shown that allergens are an important risk factor for the development of respiratory symptoms and that several avoidance measures reduce allergen levels, whether this gives clinical improvement in symptoms is debatable. Many reasons could be invoked to justify this evident discrepancy. Apart from the intrinsic methodological aspects (e.g. single or combined interventions measure, population studied, severity of respiratory symptoms, outcomes, evaluated parameters, etc.), it is important to outline that a successful approach requires that the avoided allergen is the only and real factor responsible for symptoms, the patient's education and the use of a comprehensive protocol to reduce allergen exposure. Other important factors include the involvement of the patient, the relevance of other allergens/non specific agents, and exposure to sensitizing agents also outside patient's home. It is likely that the clinical phase of allergic airway disease and the degree of bronchial (and also nasal) remodelling, in each individual, represent relevant factors for the clinical outcome of allergen avoidance procedures. Since the management of respiratory allergy is a complex strategy (including drugs, allergen avoidance, immunological and educational interventions), it is difficult in real life to distinguish the efficacy of a single intervention in comparison to the others. A combined strategy is likely to produce better clinical results. PMID- 15890512 TI - Acute and chronic effects of sildenafil in patients with pulmonary arterial hypertension. AB - Sildenafil and inhaled nitric oxide (iNO) relax smooth muscle by inhibiting the degradation and stimulating the production of cyclic guanosine monophosphate, respectively. We compared the acute pulmonary vasodilator effects of sildenafil, iNO, and epoprostenol and asked whether the combination of iNO with sildenafil had additive pulmonary vasodilator effects. We assessed the effects of extended use of sildenafil in a small cohort of patients. Twenty patients with pulmonary arterial hypertension underwent an acute vasodilator trial with sildenafil (all patients), iNO and iNO plus sildenafil (11), and epoprostenol (19). We also provided sildenafil to patients who were ineligible for, or had clinical deterioration on epoprostenol, treprostinil, or bosentan. Mean+/-se pulmonary artery pressure dropped by 13+/-3%, 19+/-4%, 14+/-3%, and 26+/-4% with epoprostenol, iNO, sildenafil, and iNO+sildenafil, respectively. Cardiac index increased with epoprostenol and sildenafil. A correlation was found between the effects of iNO and epoprostenol. Nine out of ten patients who were started on long-term sildenafil treatment alone or in combination with other vasodilators had symptomatic improvement. Three died of right heart failure. In conclusion, sildenafil is a potent acute pulmonary vasodilator, an effect that is potentiated by combination with iNO. Long-term therapy of pulmonary hypertension with sildenafil alone or in combination with other agents appears to be safe and well tolerated. PMID- 15890513 TI - Detection of CFTR mutations using ARMS and low-density microarrays. AB - The amplification refractory mutation system (ARMS) is routinely used for the identification of specific mutations within genomes. This PCR-based assay, although simple, is performed at a low-throughput scale, usually requiring gel electrophoresis for the identification of specific mutations. We have applied the ARMS technology to a low-density microarray system to facilitate the needs of the medical clinic; high-throughput capabilities and ease-of-use. Mutations within the cystic fibrosis transmembrane regulator (CFTR) gene (DeltaF508, 1717-1G>A, G542X, 621+1G>T, and N1303K) were detected by multiplex-ARMS-PCR, and fragments were post-PCR labeled with Cy5. Amine-modified probes specific for both the wild type and mutant forms of each mutation site were attached to glass substrates. Following hybridization of the PCR fragments to the attached probes (in a low density microarray format), confirmation of the presence of specific sequences was achieved using a commercial scanner, as well as a fabricated low-cost fluorescent detector and applicable software. The novel combination of the ARMS and low-density microarray technologies allows for a high-throughput, simple means to rapidly identify multiple known mutations for many genetic diseases including cystic fibrosis. PMID- 15890514 TI - Gene array analysis of the effects of chronic adrenocorticotropic hormone in vivo on immature rat adrenal glands. AB - Development of a mature adrenocortical phenotype is a critical event in the transition of mammals from fetal to postnatal life. We previously reported that the functional maturation of the adrenal glands of newborn rats is accelerated by adrenocorticotropic hormone (ACTH). We report here that chronic exposure of neonatal/juvenile rat pups to ACTH in vivo results in significant changes in expression of over 200 genes in the adrenal glands. ACTH significantly upregulated genes associated with cell signaling, gene transcription, cell migration and tissue remodeling. In addition, ACTH significantly downregulated several genes associated with de novo cholesterol biosynthesis and cholesterol trafficking. Finally, ACTH upregulated genes associated with intracellular metabolism and inactivation of glucocorticoids. The results demonstrate that the developmental effects of ACTH alter expression of a broad range of genes involved not solely in steroid synthesis, but in cellular functions related to growth and differentiation of the glands. In addition, the negative effects of ACTH on genes required for cholesterol synthesis and production of active glucocorticoids, suggests a mechanism whereby excessive production of glucocorticoids, which may have deleterious actions on developing structures like the central nervous system, is prevented. PMID- 15890515 TI - 4,5-Disubstituted cis-pyrrolidinones as inhibitors of type II 17beta hydroxysteroid dehydrogenase. Part 2. SAR. AB - 4,5-Disubstituted cis-pyrrolidinones were investigated as inhibitors of type II 17beta-hydroxysteroid dehydrogenase (17beta-HSD). Early structure-activity relationship patterns for this class of compounds are discussed. PMID- 15890516 TI - Synthesis and antioxidant, anti-inflammatory and gastroprotector activities of anethole and related compounds. AB - Some derivatives of trans-anethole [1-methoxy-4-(1-propenyl)-benzene] (1) were synthesized, by introducing hydroxyl groups in the double bond of the propenyl moiety. Two types of reactions were performed: (i) oxymercuration/demercuration that formed two products, the mono-hydroxyl derivative, 1-hydroxy-1-(4 methoxyphenyl)-propane (2) and in lesser extent the dihydroxyl derivative, 1,2 dihydroxy-1-(4-methoxyphenyl)-propane (3) and (ii) epoxidation with m chloroperbenzoic acid that also led to the formation of two products, the dihydroxyl derivative (3) and the correspondent m-chloro-benzoic acid mono-ester, 1-hydroxy-1(4-methoxyphenyl)-2-m-chlorobenzoyl-propane (4). The structures of these compounds were confirmed mainly by mass, IR, 1H and 13C NMR spectral data. The activity of anethole and hydroxylated derivatives was evaluated using antioxidant, anti-inflammatory and gastroprotector tests. Compounds (2) and (3) were more active antioxidant agents than (1) and (4). In the anti-inflammatory assay, anethole showed lower activity than hydroxylated derivatives. Anethole and in lesser extent its derivatives 2 and 4 showed significant gastroprotector activity. All tested compounds do not alter significantly the total number of white blood cells. PMID- 15890517 TI - Huntingtin is cleaved by caspases in the cytoplasm and translocated to the nucleus via perinuclear sites in Huntington's disease patient lymphoblasts. AB - Accumulation of mutant Huntingtin (Htt), especially the N-terminal-cleaved Htt, participates in the pathophysiology of Huntington's disease (HD). It is difficult to elucidate temporal properties of the translocation of "endogenous" Htt using autopsy HD patient brains. Thus, we examined the cell biology of "endogenous" Htt cleavage and nuclear translocation in cultured lymphoblasts of HD patients and controls. Apoptotic stimulation of lymphoblasts elicits caspase-dependent cleavage and selective nuclear translocation of N-terminal portions of Htt. Discrete clusters of the N-terminal Htt accumulate at unique perinuclear sites prior to nuclear translocation. Our findings suggest that caspase cleavage of Htt is cytoplasmic and precedes sorting to specific perinuclear sites followed by nuclear translocation in HD patient tissue. PMID- 15890518 TI - Squash xylem sap has activities that inhibit proliferation and promote the elongation of tobacco BY-2 cell protoplasts. AB - To elucidate the physiological functions of the substances in xylem sap, we analyzed the biological activities of xylem sap from squash (Cucurbita maxima Duch.) root using tobacco BY-2 (Nicotiana tabacum L. cv. Bright Yellow 2) cell protoplasts. When BY-2 cell protoplasts were cultivated with the total substance of squash xylem sap, the protoplasts elongated remarkably, and cell division was inhibited. Although trans-zeatin riboside (ZR), the most abundant cytokinin in squash xylem sap, had a concentration-dependent effect similar to that of total squash xylem sap, ZR concentrations several orders of magnitude greater than those found endogenously in squash xylem sap (i.e. 2 x 10(-8) M) were required to affect the growth of BY-2 cell protoplasts. The ability to stimulate cell elongation and inhibit cell division in BY-2 cell protoplasts was observed for the ethyl acetate phase fraction (pH 2) of squash xylem sap and an acetonitrile eluate fraction from reverse-phase chromatography. The xylem sap also showed inhibitory activity for auxin-induced elongation of excised cucumber hypocotyls. These results suggest that an organic substance other than ZR is produced in the root and transported to above-ground organs through the xylem via the transpiration stream, where it is involved in regulating cell proliferation and elongation in the shoot, possibly as an auxin antagonist. PMID- 15890519 TI - Induction of oxidative stress and antioxidative mechanisms in Phaseolus vulgaris after Cd application. AB - Oxidative stress has been shown to be of great importance in the toxicity of several metals (copper, zinc, ...). In this study, the relationship of cadmium phytotoxicity and antioxidative reactions in bean (Phaseolus vulgaris L.) plants was investigated. Eleven-day-old seedlings were exposed to an environmentally realistic concentration of cadmium (2 microM CdSO(4)). Several biochemical and physiological parameters were influenced even by these low concentrations. At the biochemical level, the antioxidative defence mechanism was significantly activated after 24 h of cadmium exposure. Some enzymes able of quenching reactive oxygen species (syringaldazine peroxidase, EC 1.11.1.7; guaiacol peroxidase, EC 1.11.1.7) as well as enzymes important in the reduction of NAD(P)(+) (isocitrate dehydrogenase, EC 1.1.1.42; malic enzyme, EC 1.1.1.40) were significantly elevated by cadmium exposure. Furthermore, the ascorbate-glutathione cycle appeared to be a very important mechanism against cadmium-induced oxidative stress. In leaves, significant increases of ascorbate peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2) and significant changes in the ascorbate and glutathione pool were observed. Morphological and other biochemical parameters (lipid peroxidation) were significantly enhanced 48 h after the start of the cadmium exposure. At the end of the experiment (72 h after the start of the metal treatment), even visual effects, such as chlorosis, were observed. The present data indicate that cadmium, like other metals, induces cellular redox disequilibrium suggesting that an environmentally realistic concentration of cadmium can cause oxidative stress. PMID- 15890520 TI - Cloning of a cryptochrome homologue from the holoparasitic plant Orobanche minor Sm. AB - Orobanche minor is a non-photosynthetic root holoparasitic plant. Although it is known that photosynthesis-related genes are inactivated or have been eliminated from the plastid genomes of holoparasites, little is known about the alterations in their genes involved in the signaling networks by which light regulates photosynthesis. Cryptochromes (crys), which are blue-light receptors, appear to control both photosynthesis-related and non-photosynthetic responses to light in higher plants. Because we are interested in to what extent a cry-mediated light signaling network remains in the holoparasites, we cloned CRY homologous cDNA from O. minor (OmCRY1) and used real-time RT-PCR to compare its expression under natural daylight and darkness. We found that the OmCRY1 has a high degree of homology with CRY1 s from photosynthetic plants. Expression of the OmCRY1 gene was higher in plants grown in the dark than that in the plants grown under natural daylight. This is the first report of the gene expression of a blue-light receptor in non-photosynthetic plants. PMID- 15890521 TI - Herbicidal and antioxidant responses of transgenic rice overexpressing Myxococcus xanthus protoporphyrinogen oxidase. AB - We analyzed the herbicidal and antioxidant defense responses of transgenic rice plants that overexpressed the Myxococcus xanthus protoporphyrinogen oxidase gene. Leaf squares of the wild-type incubated with oxyfluorfen were characterized by necrotic leaf lesions and increases in conductivity and malonyldialdehyde levels, whereas transgenic lines M4 and M7 did not show any change with up to 100 microM oxyfluorfen. The wild-type had decreased F(v)/F(m) and produced a high level of H(2)O(2) at 18 h after foliar application of oxyfluorfen, whereas transgenic lines M4 and M7 were unaffected. In response to oxyfluorfen, violaxanthin, beta carotene, and chlorophylls (Chls) decreased in wild-type plants, whereas antheraxanthin and zeaxanthin increased. Only a slight decline in Chls was observed in transgenic lines at 48 h after oxyfluorfen treatment. Noticeable increases of cytosolic Cu/Zn-superoxide dismutase, peroxidase isozymes 1 and 2, and catalase were observed after at 48 h of oxyfluorfen treatment in the wild type. Non-enzymatic antioxidants appeared to respond faster to oxyfluorfen induced photodynamic stress than did enzymatic antioxidants. Protective responses for the detoxification of active oxygen species were induced to counteract photodynamic stress in oxyfluorfen-treated, wild-type plants. However, oxyfluorfen-treated, transgenic plants suffered less oxidative stress, confirming increased herbicidal resistance resulted from dual expression of M. xanthus Protox in chloroplasts and mitochondria. PMID- 15890522 TI - Mg-dechelation activity in radish cotyledons with artificial and native substrates, Mg-chlorophyllin a and chlorophyllide a. AB - The Mg-dechelation activity in extracts from radish (Raphanus sativus L.) cotyledons was investigated using an artificial substrate, Mg-chlorophyllin a (Chlin) and the native substrate, chlorophyllide a (Chlide). In addition to a known a small molecular weight metal-chelating substance (MCS), Mg-releasing protein (MRP) was present when Chlin was used as the substrate. However, only MCS had Mg-dechelation activity with the native substrate. To examine the possibility of the dissociation of MRP into a protein moiety and a small molecular mass compound with an activity like MCS, extraction with low and high ionic strength buffers was carried out. No evidence was obtained that MCS is a moiety of MRP, however. Inhibitor studies showed that MCS and MRP had different susceptibilities to the inhibitors, especially to the chelators tiron and EDTA when Chlin was used as the substrate. Tiron had no effect on MRP, but it severely reduced MCS activity in both substrates. The activity of MRP increased during senescence, indicating the induction of MRP, while the activity of MCS was almost unchanged. These results suggest different reaction mechanisms by independent compounds. These findings suggest that MRP and MCS are present independently, and MCS is postulated to be a substance that catalyzes the Mg-dechelation reaction in the breakdown pathway of Chl, although MCS was not induced during senescence. The properties of MRP and MCS in relation to the small molecular mass substance obtained from strawberry fruit are also discussed. PMID- 15890523 TI - Sensitive detection of transglycosylating activity of xyloglucan endotransglycosylase/hydrolase (XTH) after isoelectric focusing in polyacrylamide gels. AB - The paper describes a sensitive and rapid zymogram technique for detection of transglycosylating activity (XET) of xyloglucan endotransglycosylase/hydrolase (XTH; EC 2.4.1.207) in polyacrylamide isoelectric focusing gels. After the electrophoresis, the separating gel was overlaid and incubated with an agarose detection gel containing XET substrates: tamarind-seed xyloglucan as the glycosyl donor and sulphorhodamine-labeled xyloglucan-derived oligosaccharides (XGO-SRs) as the glycosyl acceptors. The transglycosylation catalyzed by XTH caused incorporation of the fluorescent label into the high-M(r) polysaccharide. Selective removal of unreacted XGO-SRs from the agarose replicas by washing with organic solvents revealed the zones corresponding to XET activity as bright pink fluorescent spots under UV-light. The method appears suitable for a number of purposes such as analysis of the isoenzyme composition of XTHs with XET activity in crude extracts from various plants and plant organs, monitoring the enzyme expression at various stages of plant development and/or for checking enzyme purity in the course of its isolation procedure. PMID- 15890524 TI - Consolidation and maintenance treatments for patients with advanced epithelial ovarian cancer in complete response after first-line chemotherapy: a review of the literature. AB - Most patients with advanced epithelial ovarian cancer experience objective responses to paclitaxel/platinum-based chemotherapy, but responses are generally short-lived and the clinical outcome is still unsatisfactory. Therefore, the strategy to consolidate and to prolong the duration of response is very attractive. Different consolidation or maintenance treatments have been attempted, such as whole abdomen radiotherapy, intraperitoneal chromic phosphate, radioimmunotherapy, intraperitoneal chemotherapy, high-dose chemotherapy with haematopoietic support, prolonged administration of the first-line regimen, second-line single-agent chemotherapy, and biological agents. Clinical studies have given conflicting, inconclusive, and generally disappointing results. A recent US randomised trial appeared to show that the prolonged administration of single-agent paclitaxel (175 mg/m2 every 3 weeks) significantly improved the progression-free survival of complete responders to paclitaxel/platinum-based chemotherapy. Alternative less toxic, and probably more effective schedules of administration of chemotherapy (i.e. weekly paclitaxel) might assure a better balance between quality of life and anti-tumor activity in patients previously exposed to chemotherapy. PMID- 15890525 TI - Colorectal cancer and antiangiogenic therapy: what can be expected in clinical practice? AB - Angiogenesis is a strongly regulated process, which is dependent upon a complex interplay between inhibitory and stimulatory angiogenic factors. It is essential for tumor growth and metastasis: increased angiogenesis is correlated with poor prognosis in cancer patients. Many novel compounds that potently inhibit formation of neoplastic blood vessels have been recently developed. Major categories of angiogenesis antagonists include protease inhibitors, direct inhibitors of endothelial cell proliferation and migration, angiogenic growth factor suppressors, inhibitors of endothelial-specific integrin/survival signalling, copper chelators, and inhibitors with other specific mechanisms. There is increasing interest in developing angio-suppressive agents for colorectal cancer treatment. Some 20 direct and indirect antiangiogenesis drugs are currently being evaluated in clinical trials in colorectal cancer (CRC). Promising results have been reported. These include an increase in overall survival and reduction in the risk of death (Bevacizumab), reversal of cellular resistance (Cetuximab) and activity as second-line therapy in patients who have exhausted other available treatment options (Cetuximab, ABX-EGF, PTK-787, Gefitinib, Erlotinib). This review will outline the mechanisms of action of the principal antiangiogenic drugs, summarize the available data on the use of these new drugs in colorectal cancer, discuss their impact in clinical practice and offer a glimpse into how antiangiogenetic therapy will be integrated into the future care of patients with gastrointestinal cancers. PMID- 15890526 TI - Drug-drug interactions in oncology: why are they important and can they be minimized? AB - Adverse drug-drug interactions are a major cause of morbidity and mortality. Cancer patients are at particularly high risk of such interactions because they commonly receive multiple medications, including cytotoxic chemotherapy, hormonal agents and supportive care drugs. In addition, the majority of cancer patients are elderly, and so require medications for co-morbid conditions such as cardiovascular, gastrointestinal, and rheumatological diseases. Furthermore, the age-related decline in hepatic and renal function reduces their ability to metabolize and clear drugs and so increases the potential for toxicity. Not all drug-drug interactions can be predicted, and those that are predictable are not always avoidable. However, increased awareness of the potential for these interactions will allow healthcare providers to minimize the risk by choosing appropriate drugs and also by monitoring for signs of interaction. This review considers the basic principles of drug-drug interactions, and presents specific examples that are relevant to oncology. PMID- 15890527 TI - Caffeic acid phenethyl ester (CAPE) supplemented St. Thomas' hospital cardioplegic solution improves the antioxidant defense system of rat myocardium during ischemia-reperfusion injury. AB - BACKGROUND AND AIM OF STUDY: Cardioplegic arrest remains the method of choice for myocardial protection in cardiac surgery. Caffeic acid phenethyl ester (CAPE) prevents lipid peroxidation induced by ischemia-reperfusion injury and has a potent antioxidant property. We investigated the advantages of CAPE supplemented cardioplegic solution (St. Thomas' Hospital cardioplegic solution No.: 2) on the antioxidant defense system of myocardium against ischemia-reperfusion injury. MATERIAL AND METHODS: Isolated rat hearts were mounted on a nonrecirculating type of Langendorff apparatus. The hearts were arrested for 60 min with cardioplegic solution given at 20-min intervals and then reperfused for 15 min. The hearts were divided into three groups. Cold saline (0.9%, 4 degrees C) in group 1, St. Thomas' Hospital solution in group 2 and CAPE added St. Thomas' Hospital solution in group 3 were used as the cardioplegic solution. Krebs-Henseleit buffer solution was used for reperfusion. The tissues were examined biochemically for oxidative stress. RESULTS: Significant differences among the three groups existed in tissue myeloperoxidase (MPO), catalase (CAT), Na+-K+ ATPase activity and in the concentrations of malonydealdehyde (MDA) and 3-nitrotyrosine (3-NT). Group 2 showed significant changes in MPO (P = 0.04), Na+-K+ ATPase enzyme activity (P = 0.02) and the levels of MDA (P = 0.004) and 3-NT (P = 0.01) in comparison with group 1. Group 3 efficiently reduced MDA levels (P = 0.004) and also led to significant decrease in levels of MPO (P = 0.006), 3-NT (P = 0.01) and Na+-K+ ATPase activity (P = 0.01) and increase in the level of CAT (P = 0.004) in comparison with group 1. Significant changes were also found in the levels of MDA (P = 0.03), MPO (P = 0.04) and CAT (P = 0.009) in comparison between groups 2 and 3. CONCLUSIONS: We demonstrated that the administration of CAPE into cardioplegic solutions improves the antioxidant defense system of rat heart during the ischemia-reperfusion injury. PMID- 15890528 TI - Activation of microglia by aggregated beta-amyloid or lipopolysaccharide impairs MHC-II expression and renders them cytotoxic whereas IFN-gamma and IL-4 render them protective. AB - 'Protective autoimmunity' refers to a well-controlled anti-self response that helps the body resist neurodegeneration. The response is mediated by autoimmune T cells, which produce cytokines and growth factors. Using an in vitro assay of hippocampal slices, we show that the cytokines interferon-gamma and (especially) interleukin-4, characteristic of pro-inflammatory and anti-inflammatory T cells, respectively, can make microglia neuroprotective. Aggregated beta-amyloid, like bacterial cell wall-derived lipopolysaccharide, rendered the microglia cytotoxic. Cytotoxicity was correlated with a signal transduction pathway that down regulates expression of class-II major histocompatibility proteins (MHC-II) through the MHC-II-transactivator and the invariant chain. Protection by interleukin-4 was attributed to down-regulation of tumor necrosis factor-alpha and up-regulation of insulin-like growth factor I. These findings suggest that beneficial or harmful expression of the local immune response in the damaged CNS depends on how microglia interpret the threat, and that a well-regulated T-cell mediated response enables microglia to alleviate rather than exacerbate stressful situations in the CNS. PMID- 15890529 TI - Transmission electron microscopy characterization of macromolecular domain cavities and microstructure of single-crystal calcite tooth plates of the sea urchin Lytechinus variegatus. AB - The calcite plates and prisms in Lytechinus variegatus teeth form a complex biocomposite and employ a myriad of strengthening and toughening strategies. These crystal elements have macromolecule-containing internal cavities that may act to prevent cleavage. Transmission electron microscopy employing a small objective aperture was used to quantify several characteristics of these cavities. Cavity diameters ranged from 10 to 225 nm, the mean cavity diameter was between 50 and 60 nm, and cavities comprised approximately 20% of the volume of the crystal. Some cavities exhibited faceting and trace analysis identified these planes as being predominately of {1014} type. Through focus series of micrographs show the cavities were homogeneously distributed throughout the foil. The electron beam decomposed a substance within cavities and this suggests that these cavities are filled with a hydrated organic phase. PMID- 15890530 TI - Automated molecular microscopy: the new Leginon system. AB - We report here on the current state of our efforts in automated molecular microscopy. Our primary automated data acquisition software system, Leginon, has been completely redesigned over the past two years. The new distributed system has been developed using the Python programming language and is compatible with both Linux and Windows operating systems. The new flexible architecture was designed to allow for the development of customized data collection protocols, several of which are described here. The system has been used to acquire data for approximately 150 experiments and we have demonstrated the capacity for high throughput data acquisition by acquiring images of more than 100,000 particles in a single session at the microscope. PMID- 15890531 TI - The use of immunostimulants in fish larval aquaculture. AB - The production of fish larvae is often hampered by high mortality rates, and it is believed that most of this economic loss due to infectious diseases is ca. 10% in Western European aquaculture sector. The development of strategies to control the pathogen load and immuno-prophylactic measures must be addressed further to realise the economic "potential" production of marine fish larvae and thus improve the overall production of adult fish. The innate defence includes both humoral and cellular defence mechanisms such as the complement system and the processes played by granulocytes and macrophages. A set of different substances such as beta-glucans, bacterial products, and plant constituents may directly initiate activation of the innate defence mechanisms acting on receptors and triggering intracellular gene activation that may result in production of anti microbial molecules. These immunostimulants are often obtained from bacterial sources, brown or red algae and terrestrial fungi are also exploited as source of novel potentiating substances. The use of immunostimulants, as dietary supplements, can improve the innate defence of animals providing resistance to pathogens during periods of high stress, such as grading, reproduction, sea transfer and vaccination. The immunomodulation of larval fish has been proposed as a potential method for improving larval survival by increasing the innate responses of the developing animals until its adaptive immune response is sufficiently developed to mount an effective response to the pathogen. To this end it has been proposed that the delivery of immunostimulants as a dietary supplement to larval fish could be of considerable benefit in boosting the animals innate defences with little detriment to the developing animal. Conversely, there is a school of thought that raises the concern of immunomodulating a neotanous animal before its immune system is fully formed as this may adversely affect the development of a normal immune response. PMID- 15890532 TI - Phylogeny and ontogeny of fish leucocytes. AB - In contrast to higher vertebrates, most fish species hatch at the embryonic stage of life. Consequently, they have to defend against a variety of micro-organisms living in their aquatic environment. This paper is focussed on the development of leucocytes functioning within this early innate system and later on in the acquired immune system (B and T cells). Most of the data are derived from cyprinid fish (zebrafish, carp), which are excellent models to study early ontogeny. Attention is also paid to the phylogeny of leucocytes, with special attention to early chordates. It is clear that young fish use innate mechanisms during the first weeks/months of their development. In zebrafish, a variety of hematopoietic genes have been sequenced which allow a detailed picture of the development of the distinct leucocytes and their precursors. In cyprinids and sea bass, the thymus is the first lymphoid organ and T cells appear to be selected there much earlier than the first detection of T cell-dependent antibody responses. The first B cells are most probably generated in head kidney. Although T cells are selected earlier than B cells, T cell independent responses occur earlier than the T cell-dependent responses. The very early (pre-thymic) appearance of T-like cells in gut of sea bass and carp suggests an extra-thymic origin of these cells. However, B cells populate the GALT much later than spleen or kidney, indicating a rather late appearance of mucosal immunity. The first plasma cells are found long after the intake of food in cyprinids, but in many marine fish they appear around the first food intake. In general, acquired immunity is not correlated to food intake. PMID- 15890533 TI - Comparative organ differentiation during early life stages of marine fish. AB - The basic developmental mechanisms of teleosts are similar, but there are differences with respect to the timing of developmental events. These events are controlled by genetic and environmental factors. Direct comparisons of organogenesis are complicated due to large variations in egg sizes and incubation temperatures between species. But in general, cultivated small marine pelagic fish larvae originating from rather small eggs (like gadoids, flatfishes, sparids) hatch with a relatively large yolk sac, a larval finfold and subdermal space and under-developed organs. Developmental status at hatch differs between species and the duration of the yolk sac period varies. Main organs and organ systems become functional by first feeding and differentiate during the larval stage and metamorphosis. Species developing directly via large yolk-rich eggs and a long incubation period have a juvenile like morphology and organ functionality at first feeding, sometimes immediately after hatch (like wolffishes). Histomorphological and cell- or organ functional studies of developing embryos and larvae of cultivated species constitute basic information for understanding species-specific events, of utmost importance for improving production protocols. Information is still lacking on early functionality of endocrine and immunocompetent tissues and organs, areas that deserve future focus. PMID- 15890534 TI - The neural bases of amusement and sadness: a comparison of block contrast and subject-specific emotion intensity regression approaches. AB - Neuroimaging studies have made substantial progress in elucidating the neural bases of emotion. However, few studies to date have directly addressed the subject-specific, time-varying nature of emotional responding. In the present study, we employed functional magnetic resonance imaging to examine the neural bases of two common emotions--amusement and sadness--using both (a) a stimulus based block contrast approach and (b) a subject-specific regression analysis using continuous ratings of emotional intensity. Thirteen women viewed a set of nine 2-min amusing, sad, or neutral film clips two times. During the first viewing, participants watched the film stimuli. During the second viewing, they made continuous ratings of the intensity of their own amusement and sadness during the first film viewing. For sad films, both block contrast and subject specific regression approaches resulted in activations in medial prefrontal cortex, inferior frontal gyrus, superior temporal gyrus, precuneus, lingual gyrus, amygdala, and thalamus. For amusing films, the subject-specific regression analysis demonstrated significant activations not detected by the block contrast in medial, inferior frontal gyrus, dorsolateral prefrontal cortex, posterior cingulate, temporal lobes, hippocampus, thalamus, and caudate. These results suggest a relationship between emotion-specific temporal dynamics and the sensitivity of different data analytic methods for identifying emotion-related neural responses. These findings shed light on the neural bases of amusement and sadness, and highlight the value of using emotional film stimuli and subject specific continuous emotion ratings to characterize the dynamic, time-varying components of emotional responses. PMID- 15890535 TI - Coronavirus phylogeny based on a geometric approach. AB - A novel coronavirus has been identified as the cause of the outbreak of severe acute respiratory syndrome (SARS). Previous phylogenetic analyses based on sequence alignments show that SARS-CoVs form a new group distantly related to the other three groups of previously characterized coronaviruses. In this paper, a geometric approach based on the Z-curve representation of the whole genome sequence is proposed to analyze the phylogenetic relationships of coronaviruses. The evolutionary distances are obtained through measuring the differences among the three-dimensional Z-curves. The Z-curve is approximately described by its geometric center and the associated three eigenvectors, which indicate the center position and the trend of the Z-curve, respectively. Although some information is lost due to the approximate description of the Z-curve, the phylogenetic tree constructed based on these parameters is consistent with those of previous analyses. The present method has the merits of simplicity and intuitiveness, but it is still in its premature stage. Because the phylogenetic relationships are inferred from the whole genome, instead of some individual genes, the present method represents a new direction of phylogeny study in the post-genome era. PMID- 15890536 TI - Molecular systematics of the gonorynchiform fishes (Teleostei) based on whole mitogenome sequences: implications for higher-level relationships within the Otocephala. AB - Although the order Gonorynchiformes includes only 31 species assigned to seven genera and four families, it exhibits a large variety of anatomical structures, making difficult the reconstruction of phylogenetic relationships among its representatives. Within the basal teleosts, the Gonorynchiformes belong to the Otocephala where they have been alternatively placed as the sister group of the Otophysi and of the Clupeiformes. In this context, we investigated the phylogeny of the Gonorynchiformes using whole mitogenome sequences from 40 species (six being newly determined for this study). Our taxonomic sampling included at least one species of each gonorynchiform genus and of each other major otocephalan lineage. Unambiguously aligned, concatenated mitogenomic sequences (excluding the ND6 gene and control region) were divided into five partitions (1st, 2nd, and 3rd codon positions, tRNA genes, and rRNA genes) and partitioned Bayesian analyses were conducted. The resultant phylogenetic trees were fully resolved, with most of the nodes well supported by the high posterior probabilities. As expected, the Otocephala were recovered as monophyletic. Within this group, the mitogenome data supported the monophyly of Alepocephaloidei, Gonorynchiformes, Otophysi, and Clupeiformes. The Gonorynchiformes and the Otophysi formed a sister group, rending the Ostariophysi monophyletic. This result conflicts with previous mitogenomic phylogenetic studies, in which a sister relationship was found between Clupeiformes and Gonorynchiformes. We discussed the possible causes of this incongruence. Within the Gonorynchiformes, the following original topology was found: (Gonorynchus (Chanos (Phractolaemus (Cromeria (Grasseichthys (Kneria, Parakneria)))))). We confirmed that the paedomorphic species Cromeria nilotica and Grasseichthys gabonensis belong to the family Kneriidae; however, the two species together did not form a monophyletic group. This result challenges the value of reductive or absent characters as synapomorphies in this group. PMID- 15890537 TI - Phylogeny of endosymbiont bacteria harbored by the woodroach Cryptocercus spp. (Cryptocercidae: Blattaria): molecular clock evidence for a late Cretaceous- early Tertiary split of Asian and American lineages. PMID- 15890538 TI - Prolonged Ca2+ transients in ATP-stimulated endothelial cells exposed to 50 Hz electric fields. AB - Human umbilical vein endothelial cells were exposed to sinusoidal electric fields of 0.3 or 30 kV/m, 50 Hz, for 24 h. Changes in intracellular calcium concentration ([Ca(2+)](i)) induced by ATP-stimulation in the absence of extracellular Ca(2+) were observed in individual cells. No differences were observed between the exposure and sham-exposure groups in [Ca(2+)](i) resting level before ATP-stimulation, or in the [Ca(2+)](i) peak levels induced by stimulation. However, the duration of the initial transients in [Ca(2+)](i) following an ATP stimulus was significantly prolonged by exposure to a 30 kV/m field. The inositol trisphosphate receptor inhibitor, xestospongin C, inhibited the ATP-induced elevation in [Ca(2+)](i) in both the exposure and sham-exposure groups. The ATP-receptor P2Y appeared to play an important role in the increase of [Ca(2+)](i). The present results suggest that an extremely low-frequency electric field affects the function of vascular endothelial cells by a mechanism involving activation of P2Y. PMID- 15890539 TI - A tentative classification of centrosome abnormalities in cancer. AB - Centrosome anomalies are detected in virtually all human cancers. They have been implicated in multipolar mitoses, chromosome missegregation, and genomic instability. Despite extensive studies on the type and frequency of centrosome anomalies, a causative relationship between centrosome aberrations and chromosomal instability has been difficult to establish. For example, centrosome amplification can be present without associated chromosomal instability. In addition, not all cells appear to be permissive for centrosome-related mitotic defects suggesting that cellular mechanisms that limit the harmful effects of spindle malformation on genome integrity may exist. This review proposes to classify centrosome abnormalities in tumor cells into three groups based on their relevance to genomic instability: primary centrosome overduplication, transient centrosome accumulation, and permanent centrosome accumulation. Whereas the first two categories are associated with an increased risk of chromosomal missegregation, the latter category may not contribute to the propagation of genomic instability. Therefore, centrosome anomalies should not per se be viewed as a universal cause of chromosomal instability, rather, they need to be assessed in the cellular context in which they occur. PMID- 15890540 TI - Memory enhancement by a semantically unrelated emotional arousal source induced after learning. AB - It has been well established that moderate physiological or emotional arousal modulates memory. However, there is some controversy about whether the source of arousal must be semantically related to the information to be remembered. To test this idea, 35 healthy young adult participants learned a list of common nouns and afterward viewed a semantically unrelated, neutral or emotionally arousing videotape. The tape was shown after learning to prevent arousal effects on encoding or attention, instead influencing memory consolidation. Heart rate increase was significantly greater in the arousal group, and negative affect was significantly less reported in the non-arousal group after the video. The arousal group remembered significantly more words than the non-arousal group at both 30 min and 24 h delays, despite comparable group memory performance prior to the arousal manipulation. These results demonstrate that emotional arousal, even from an unrelated source, is capable of modulating memory consolidation. Potential reasons for contradictory findings in some previous studies, such as the timing of "delayed" memory tests, are discussed. PMID- 15890541 TI - Factors affecting the long-term patency of crossover femorofemoral bypass graft. AB - OBJECTIVES: To determine the factors affecting long-term patency of crossover femorofemoral bypass (FFB) graft. DESIGN: A retrospective clinical study of a prospectively registered database. MATERIALS: Two hundred and sixteen FFBs performed for 192 patients with atherosclerotic iliac occlusive disease. METHODS: The clinical and surgical variables influencing graft patency were assessed with log-rank test and Cox's proportional hazard analysis. RESULTS: The primary patency rates of all FFB grafts at 3 and 5 years were 73+/-4 and 65+/-5%, respectively. By multivariate analysis, hypertension (Odds ratio 2.8, P=0.002) and critical ischemia (Odds ratio 0.42, P=0.01) significantly (P<.05) influenced long-term patency of FFB grafts. CONCLUSION: The long-term patency of FFB grafts was not affected by procedural modifications. Graft patency was inferior in patients with severe lower limb ischemia but superior in patients with hypertension. Further study is required to clarify the mechanism of an unexpected beneficial effect of hypertension on FFB graft patency. PMID- 15890542 TI - Transluminal coil embolization of an inferior gluteal artery aneurysm by ultrasound-guided direct puncture of the target vessel. AB - Pseudoaneurysms of the inferior gluteal artery are rare. We describe a case of an inferior gluteal pseudoaneurysm that presented as a painful mass in the buttock. A percutaneous thrombin injection under ultrasound guidance failed to occlude the sac, probably due to the wide neck of the aneurysm. Subsequently, transluminal coil embolization by ultrasound-guided direct puncture of the inferior gluteal artery achieved complete thrombosis of the sac. Ultrasound-guided coil embolization is recommended in the treatment of peripheral aneurysms where catheter placement using conventional interventional procedures is difficult. PMID- 15890543 TI - Re: Emboli rate during an early after carotid endarterectomy after a single preoperative dose of 120 mg acetylsalicylic acid--a prospective double-blind placebo controlled randomized trial by Tytgat et al. Eur J Vasc Endovasc Surg 2005;29:156-161. PMID- 15890544 TI - Determinants of failure of brachiocephalic elbow fistulas for haemodialysis. AB - OBJECTIVES: The aim of this study was to analyse the results of brachiocephalic fistulas for haemodialysis and to determine possible predictors of failure. PATIENTS AND METHODS: Between April 1999 and September 2004, a consecutive series of 100 autologous brachiocephalic fistulas were created in 96 patients. There were 57 men and 39 women with a mean (SD) age of 59.2 (15.6) years. Data were prospectively gathered. RESULTS: The mean (SD) follow-up was 20.1 (16.4) months. The primary, primary assisted, and secondary patency rates after 6 months were 73.4, 83.2 and 86.4%, respectively. After 1 year, these figures were 54.7, 72.3 and 79.2%, and after 2 years 40.4, 59.2 and 67.5%, respectively. Predictors of failure with regard to primary patency, determined with Cox regression multivariate analysis, included diabetes mellitus (HR 2.81, p < 0.001) and a history of contralateral PTFE loop graft (HR 7.79, p = 0.007). CONCLUSION: Primary patency of brachiocephalic fistulas is comparable to that of radiocephalic fistulas. Primary assisted and secondary patency rates can, however, be brought to a much higher level, especially in patients without diabetes and a large-diameter venous outflow tract. PMID- 15890545 TI - Improvement of the walking ability in intermittent claudication due to superficial femoral artery occlusion with supervised exercise and pneumatic foot and calf compression: a randomised controlled trial. AB - OBJECTIVES: To compare the effect of unsupervised exercise, supervised exercise and intermittent pneumatic foot and calf compression (IPC) on the claudication distance, lower limb arterial haemodynamics and quality of life of patients with intermittent claudication. METHODS: Thirty-four eligible patients with stable intermittent claudication were randomised to IPC (n = 13, 3h/d for 6 months), supervised exercise (n = 12, three hourly sessions/week for 6 months) or unsupervised exercise (n = 9). In each patient, initial claudication distance (ICD), absolute claudication distance (ACD), resting ankle brachial pressure index (ABPI), and resting hyperaemic calf arterial inflow were measured before, 6 weeks, 6 months and 1 year after randomisation. Quality of life was assessed with the short form (SF)-36, walking impairment (WIQ) and intermittent claudication questionnaires (ICQ). RESULTS: Compared with unsupervised exercise, both IPC and supervised exercise, increased ICD and ACD, up to 2.83 times. IPC increased arterial inflow (p < 0.05 at 6 weeks) and ABPI. Supervised exercise decreased arterial inflow and increased ABPI (p < 0.05 at 6 months). Unsupervised exercise had no effect on arterial inflow or ABPI. IPC improved significantly the ICQ score and the speed score of the WIQ, while supervised exercise improved the WIQ claudication severity score. At 1 year clinical effectiveness of supervised exercise and IPC was largely preserved. CONCLUSIONS: IPC, by augmenting leg perfusion, achieved improvement in walking distance comparable with supervised exercise. Long-term results in a larger number of patients will provide valuable information on the optimal treatment modality of intermittent claudication. PMID- 15890546 TI - Characteristics of peripheral microembolization during iliac stenting: Doppler ultrasound monitoring. AB - OBJECTIVE: To evaluate the characteristics of distal microembolic signals (MES) during iliac stenting using Doppler ultrasound monitoring. DESIGN: Prospective clinical study. METHODS: A 2-MHz probe was used to monitor continuously at the ipsilateral tibioperoneal trunks during technically and hemodynamically successful iliac stenting in 10 patients without infrainguinal occlusive lesion. MESs at guide-wire, balloon, or stent crossing (phase 1), predilatation (phase 2), stent deployment (phase 3), postdilatation (phase 4), and contrast medium or heparinized saline injection (at injection) were analyzed. Differentiation of gaseous emboli from particulate emboli was achieved by calculation of the sample volume length. RESULTS: No distal embolic complications were observed. Five hundred and forty-one MESs were detected. The MES incidence and intensity in phase 3 were significantly higher than those in phase 1, phase 2, and phase 4 (p<0.05). The MES intensity at injection was significantly higher than that in each of four phases (p<0.0001). CONCLUSIONS: Both the MES incidence and intensity were highest at stent deployment. Further study is required of microembolism during endovascular procedures in the lower extremities. PMID- 15890547 TI - Early and long-term results of replacement of the descending aorta. AB - OBJECTIVES: The aim of this analysis was to evaluate our results of open surgery on the descending thoracic aorta as benchmark to define indications for endovascular treatment. METHODS: Between January 1981 and December 2000, 115 patients underwent replacement of the descending or thoraco-abdominal aorta. Follow-up to 20 years was complete in 98%. RESULTS: Early mortality was 19% and paraplegia rate was 7%. Surgery before 1990 and coronary artery disease were independent predictors for early mortality. Thoraco-abdominal repair and normothermia were independent predictors for paraplegia. Ten years survival rate was 63%. CONCLUSIONS: Our results confirm that replacement of the descending aorta can be performed today with acceptable low mortality and morbidity and with consistent exclusion of the aneurysm or dissection. Long-term results of endovascular stent-grafts in the descending aorta are unclear. In our opinion endovascular stent-grafts should be reserved for high risk patients, acute dissection or acute aortic rupture. PMID- 15890548 TI - New aspects of the location of neuronal nitric oxide synthase in the skeletal muscle: a light and electron microscopic study. AB - The action of nitric oxide (NO) synthesized by NO synthases (NOS) is spatially restricted. Hence, the intracellular location of NOS might play an important role for the functional interactions of NO with its target molecules. In the skeletal muscle the neuronal NOS (nNOS) is considered to be the predominant isoform expressed as a muscle specific elongated splice variant. There are only a few and highly discrepant reports of the subcellular distribution of nNOS, which prompted us to re-examine the distribution of nNOS in the skeletal muscle of rat and mouse applying immunocytochemistry and NADPH-diaphorase (NADPH-d) histochemistry. Light microscopically, the sarcolemma, areas beneath the sarcolemma, areas around the nuclei, and the cross striation were labeled by antibodies and by the NADPH-d reaction as well. Ultrastructurally, nNOS visualized immunocytochemically or by the histochemical BSPT-reaction, was associated discretely with extrajunctional portions of the sarcolemma. Both reaction products were additionally observed in the vicinity of endoplasmic reticulum and mitochondria, or associated with their outer membranes. In the neuromuscular junction (NMJ)-region NOS was localized to the cytoplasm of nerve terminals and terminal Schwann cells. In contrast to the commonly accepted assumption, the enzyme was found in association with the presynaptic, and not with the postsynaptic membrane. Cytosolic NADPH-d was exhibited especially between mitochondria accumulated in the postsynaptic region of the NMJ. Surprisingly, in nNOS-/--mice the skeletal muscle showed patterns of significant nNOS-immunoreactivity and NADPH-d activity possibly due to alternative nNOS-splice isoforms, which might be up-regulated to compensate for decreased NO formation. PMID- 15890549 TI - The T-786C and C774T endothelial nitric oxide synthase gene polymorphisms independently affect the onset pattern of severe diabetic retinopathy. AB - Genetic factors could be implicated in the pathogenesis of severe diabetic retinopathy (DR). Recently, we reported a strong association between the eNOS4b/a endothelial nitric oxide synthase (eNOS) polymorphism and severe DR. To examine whether T-786C and C774T eNOS polymorphisms are involved in severe DR, 254 Caucasians with longstanding C-peptide-negative type 1 diabetes, 128 patients with absent/mild DR (control group), and 126 patients with preproliferative/proliferative DR (study group) were genotyped. The distribution of T-786C and C774T eNOS polymorphisms was in Hardy-Weinberg equilibrium and did not differ between the study and control groups. However, in case patients (n=126), T-786C and C774T polymorphisms influenced the onset pattern of severe DR (P=0.0169 and P=0.0257, respectively). The C-786C genotype was associated with early-onset severe DR (duration of diabetes: 15.2+/-5.9 vs. 19.4+/-6.3 years, P=0.0105), and the homozygous T774T genotype was associated with late-onset severe DR (24.3+/-7.0 vs. 18.4+/-6.2 years, P=0.0067). In the case of patients with high glycosylated hemoglobin levels (HbA1c >8%, n=88), the association between the T-786C polymorphism and early-onset severe DR was stronger (P=0.0068). Case patients carrying the C-786C genotype had higher HbA1c values (9.61+/-1.89%) than those carrying the T-786T genotype (8.93+/-1.47%, P=0.0173). Multivariate analysis showed that T-786C polymorphism was the best independent factor for onset pattern of severe DR (P<0.001). These findings, supported by previous associations between eNOS4b/a polymorphism and DR, suggest that T-786C and C774T eNOS polymorphisms affect the onset pattern of severe DR. PMID- 15890550 TI - Midostaurin upregulates eNOS gene expression and preserves eNOS function in the microcirculation of the mouse. AB - Nitric oxide (NO) derived from endothelial NO synthase (eNOS) is a powerful vasodilator and possesses vasoprotective effects. Therefore, augmentation of eNOS expression and -activity by pharmacological means could provide protection against cardiovascular disease. However, this concept has been questioned recently, because in several disease models, eNOS upregulation was associated with a dysfunctional enzyme (referred to as eNOS uncoupling). In contrast, the present study demonstrates that an eNOS gene expression-enhancing compound with additional protein kinase C (PKC) inhibitory properties can upregulate eNOS while preserving its enzymatic function. Apolipoprotein E-knockout mice were treated for 7 days with midostaurin (4'-N-benzoyl staurosporine, compound CGP 41251, 50 125 mg/kg/day), a PKC inhibitor previously shown to increase eNOS expression and NO production in cultured human endothelial cells. Midostaurin treatment enhanced eNOS mRNA expression (RNase protection assay) in mouse aorta, kidney, and heart in a dose-dependent fashion. In the dorsal skinfold microcirculation, midostaurin produced an arteriolar vasorelaxation (intravital microscopy), which could be prevented by the NOS inhibitor L-NAME, indicating that the upregulated eNOS remained functional. In organ chamber experiments, the aorta from midostaurin treated mice showed an enhanced NO-mediated relaxation in response to acetylcholine. Accordingly, serum levels of nitrite/nitrate (NO-Analyzer) were increased, and the production of reactive oxygen species in the aorta (L-012 chemiluminescence) was reduced by midostaurin. Thus, in mice in vivo, midostaurin treatment results in enhanced expression of eNOS with preserved enzyme function and enhanced production of bioactive NO. Given the beneficial effects of endothelial-derived NO, vasoprotective and anti-atherosclerotic effects are likely to ensue. PMID- 15890551 TI - Effects of mechanistically distinct NF-kappaB inhibitors on glial inducible nitric-oxide synthase expression. AB - Nuclear factor (NF)-kappaB is an important regulator of inflammatory gene expression. Transcriptional regulation of Nos2, the inducible nitric-oxide synthase (iNOS) gene, is complex and not fully understood, but appears to be regulated in part by NF-kappaB. To further understand the role of NF-kappaB in Nos2 expression, we compared three functionally distinct NF-kappaB inhibitors on NF-kappaB transactivation and iNOS induction by rat C6 glial cells. Cytokine induced activation of a consensus NF-kappaB-reporter gene was concentration dependently inhibited by BAY 11-7082, MG-132, and helenalin. The rank order of potency was MG-132>helenalin>BAY 11-7082, with low concentrations of helenalin stimulating reporter gene activity. Cytokine-stimulated iNOS expression, measured by nitrite accumulation and in vitro l-citrulline production, was similarly reduced by exposing C6 cells to the NF-kappaB inhibitors. Surprisingly, activation of Nos2-reporter gene constructs containing the proximal 188 bp (containing one kappaB site) or proximal 94 bp (no kappaB site) of the rat promoter also was inhibited with the same rank order of potency. Interestingly, low concentrations of helenalin increased activity of both promoter constructs, while BAY 11-7082 poorly inhibited the 94-bp activity. This is the first report describing BAY 11-7082 and helenalin effects on iNOS expression in astroglia. Given the reported mechanism of actions for these inhibitors, cytokine-induced glial iNOS expression appears more sensitive to disruption of proteasome degradation and p65 function than modulation of IkappaB phosphorylation. These findings may foster the design of therapeutic agents aimed at NF-kappaB associated pathways involved in neuroinflammation, especially iNOS expression. PMID- 15890552 TI - Identification and expression of a small heat shock protein in two lines of the endoparasitic wasp Venturia canescens. AB - Increased expression of small heat shock proteins (sHSPs) is known to be a key regulatory mechanism in extending tolerance to a variety of environmental stresses. In the present study, a full-length cDNA clone encoding a member of the alpha-crystallin/sHSP family was isolated and characterized from the endoparasitic wasp Venturia canescens (Hymenoptera: Ichneumonidae). Western blot analysis indicated that the mature protein has a mass of about 35 kDa (Vc_sHSP35). Sequence analysis of RT-PCR products revealed that two transcript forms of the gene are expressed in different developmental stages and tissues of the wasp, with the longer form likely to contain an unspliced intron sequence. Furthermore, gene expression was analysed in ovaries of V. canescens wasps from two genetically different lines after exposure to different temperatures (heat or cold shock and heat or cold acclimation, respectively). Wasps from both lines principally showed the same cold induced change in expression of the shorter transcript form (Vc_sHSP35-2). However, expression levels were higher in wasps from one line compared to the other. These results are discussed in relation to the environmental stress resistance and molecular ecology of both V. canescens lines. PMID- 15890553 TI - Improved protection by recombinant BCG. AB - Mycobacterium bovis bacille Calmette Guerin (BCG) is one of the most widely used live vaccines. Technologic advancement in genome manipulation enables the construction of recombinant BCG (rBCG) strains, which can be employed as highly immunogenic vaccines against tuberculosis with improved safety profile. PMID- 15890554 TI - Functional analysis of immune response genes in Drosophila identifies JNK pathway as a regulator of antimicrobial peptide gene expression in S2 cells. AB - The templates of innate immunity have ancient origins. Thus, such model animals as the fruit fly, Drosophila melanogaster, can be used to identify gene products that also play a key role in the innate immunity in mammals. We have used oligonucleotide microarrays to identify genes that are responsive to gram negative bacteria in Drosophila macrophage-like S2 cells. In total, 53 genes were induced by greater than threefold in response to Escherichia coli. The induction of all these genes was peptidoglycan recognition protein LC (PGRP-LC) dependent. Twenty-two genes including 10 of the most strongly induced genes are also known to be up-regulated by septic injury in vivo. Importantly, we identified 31 genes that are not known to respond to bacterial challenge. We carried out targeted dsRNA treatments to assess the functional importance of these gene products for microbial recognition, phagocytosis and antimicrobial peptide release in Drosophila S2 cells in vitro. RNAi targeting three of these genes, CG7097, CG15678 and beta-Tubulin 60D, caused altered antimicrobial peptide release in vitro. Our results indicate that the JNK pathway is essential for normal antimicrobial peptide release in Drosophila in vitro. PMID- 15890555 TI - Prime-boost immunisation strategies for tuberculosis. AB - Vaccines against intracellular pathogens such as Mycobacterium tuberculosis need to induce strong cellular immune responses. Heterologous prime-boost immunisation strategies induce higher levels of both CD4+ and CD8+ T cells than homologous boosting with the same vector. Recombinant pox-viruses are particularly good at boosting previously primed T cell responses. Using BCG as the priming immunisation in such a heterologous prime-boost strategy is a practical solution, which allows the beneficial effects of BCG in children to be maintained. PMID- 15890556 TI - Introduction: characterization and functions of human T regulatory cells. AB - The field of human T regulatory (Treg) cells is a rapidly progressing, but still confused field of immunology. The effects of dendritic cell (DC) manipulation in Treg generation and the main features of human "natural" Treg cells, as well as of different populations of adaptive Treg subsets, are still partially unclear. However, it is clear that Treg cells play an important role in human diseases, such as autoimmune disorders, allergy, HIV infection, tumors and graft-versus host disease. PMID- 15890557 TI - Crystal structure of Anaerobiospirillum succiniciproducens PEP carboxykinase reveals an important active site loop. AB - The 2.2 Angstroms resolution crystal structure of the enzyme phosphoenolpyruvate carboxykinase (PCK) from the bacterium Anaerobiospirillum succiniciproducens complexed with ATP, Mg(2+), Mn(2+) and the transition state analogue oxalate has been solved. The 2.4 Angstroms resolution native structure of A. succiniciproducens PCK has also been determined. It has been found that upon binding of substrate, PCK undergoes a conformational change. Two domains of the molecule fold towards each other, with the substrates and metal ions held in a cleft formed between the two domains. This domain movement is believed to accelerate the reaction PCK catalyzes by forcing bulk solvent molecules out of the active site. Although the crystal structure of A. succiniciproducens PCK with bound substrate and metal ions is related to the structures of PCK from Escherichia coli and Trypanosoma cruzi, it is the first crystal structure from this class of enzymes that clearly shows an important surface loop (residues 383 397) from the C-terminal domain, hydrogen bonding with the peptide backbone of the active site residue Arg60. The interaction between the surface loop and the active site backbone, which is a parallel beta-sheet, seems to be a feature unique of A. succiniciproducens PCK. The association between the loop and the active site is the third type of interaction found in PCK that is thought to play a part in the domain closure. This loop also appears to help accelerate catalysis by functioning as a 'lid' that shields water molecules from the active site. PMID- 15890558 TI - OpenSDE: a strategy for expressive and flexible structured data entry. AB - PURPOSE: This description focuses on the expressiveness and flexibility of OpenSDE: an application that supports recording of structured narrative data. METHODS: OpenSDE enables data entry with (customizable) forms based on trees of medical concepts. The relevant scope for data entry can be tailored per medical domain by construction of a domain-specific tree. OpenSDE is intended for structuring narrative data to make these available for both care and research. RESULTS: The OpenSDE application is currently in use at several departments in our academic hospital, including radiology, neurology, pediatrics, and child psychiatry. OpenSDE is available for all in open source. PMID- 15890559 TI - Acute effects of sediments taken from an urban stream on physiological and biochemical parameters of the neotropical fish Prochilodus lineatus. AB - Juveniles of Prochilodus lineatus were exposed to sediments collected from one of five sites along an urban stream into which various types of contaminants are discharged. After 24 or 96 h fish were examined and the results compared with those from control groups (fish exposed only to water, for the same period). Plasma ion levels varied significantly and fish exposed to site 5 sediment showed a transient increase in both sodium and chloride concentrations. Plasma glucose was significantly higher in fish exposed to sediment from sites 2 and 5. The higher liver glutathione-S-transferase activity registered in fish exposed to sediment from sites 1, 4 and 5 suggests the presence of organic contaminants at these sites and the enhancement of liver catalase activity in fish exposed to sediment from sites 3 and 4 may be due to contaminant-mediated oxyradical production. The overall results revealed that sites 4 and 5 are more severely contaminated, probably due to organic contaminants from agricultural sources and municipal landfill. PMID- 15890560 TI - Antioxidant responses of the Mediterranean mussel, Mytilus galloprovincialis, to environmental variability of dissolved oxygen. AB - Physiological responses of Mytilus galloprovincialis against environmental dissolved oxygen partial pressure (pO(2)) variation were studied in terms of the modulated induction of the main antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT) and selenium-dependent glutathione peroxidase (GPX). Field in vivo studies were performed at two sites of the Lagoon of Venice, characterized by different aquatic environmental conditions implying different pO(2). SOD and GPX are more active in gills, and their complementary role is discussed. CAT is more active in the digestive gland, where the enzyme dismutates H(2)O(2) derived from divalent reduction of O(2) performed by various oxidases in peroxisomes. Antioxidant enzyme activities are correlated with water dissolved oxygen (DO), especially in the gills. This tissue, because of its anatomical localization and its physiological role, responds to DO variations modulating the induction of the antioxidant enzymes as a protection mechanism against potential toxicity due to increases in ROS formation. PMID- 15890561 TI - The 6-min walk test (6MW) as an efficacy endpoint in pulmonary arterial hypertension clinical trials: demonstration of a ceiling effect. AB - BACKGROUND: PAH trials traditionally use 6MW as the primary endpoint. Concerns regarding a "ceiling effect" masking efficacy have led to exclusion of patients with milder disease from most trials (BL 6MW>450 m). STRIDE I evaluated the selective endothelin A receptor antagonist, sitaxsentan (SITAX), in a 12-week randomized, double-blind, trial (178 patients) employing placebo (PBO), 100 mg or 300 mg SITAX orally once daily in PAH and included patients with NYHA class II, congenital heart disease and a BL 6MW>450 m, groups often excluded from previous trials. METHODS: We analyzed 6MW effects For All Pts (intention-to treat) and those meeting Traditional enrollment criteria, defined as patients with NYHA class III or IV and 6MW< or =450 m at BL with idiopathic PAH or PAH related to connective tissue disease. The 100 mg and 300 mg SITAX arms are pooled based on similar treatment effects on 6MW. CONCLUSION: Existence of a "ceiling effect" is supported by these data. The magnitude of the treatment effect and statistical power when using 6MW as the endpoint. Comparisons between PAH trials that do not adjust for the effects of differing enrollment criteria require caution. PMID- 15890562 TI - Lactic acid buffering, nonmetabolic CO2 and exercise hyperventilation: a critical reappraisal. AB - It has been suggested that hyperventilation and the disproportionate increase in VCO2 versus VO2 above the ventilatory threshold (V(TH)) in ramp exercise are due to the production of nonmetabolic CO2 in muscle because of lactic acid buffering by plasma bicarbonate entering the cell in exchange with lactate [Wasserman, K., 1982. Dyspnea on exertion. Is it the heart or the lungs? JAMA 248, 2039-2043]. According to this model, plasma standard bicarbonate concentration decreases in a approximately 1:1 ratio with the increase in plasma lactate concentration, 1 mmol of CO2 is generated above that produced by aerobic metabolism for each mmol of lactic acid buffered, and nonmetabolic CO2 produced in the muscle is partly responsible for hyperventilation because of the resulting increase in the CO2 flow to the lungs. The present report shows that this model is not consistent with experimental data: (1) bicarbonate is not the main buffer in the muscle; (2) the decrease in standard bicarbonate concentration is not the mirror image of the increase in lactate concentration; (3) buffering by bicarbonate does not increase CO2 production in muscle (no nonmetabolic CO2 is produced in tissues); (4) the CO2 flow to the lungs, which should not be confused with VCO2 at the mouth, does not increase at a faster rate above than below V(TH). The disproportionate increase in VCO2 at the mouth above V(TH) is due to hyperventilation (not the reverse) and to the low plasma pH which both reduce the pool of bicarbonate readily available in the body. PMID- 15890563 TI - Aerosols in the study of convective acinar mixing. AB - Convective mixing (CM) refers to the different transport mechanisms except Brownian diffusion that irreversibly transfer inspired air into resident air and can be studied using aerosol bolus inhalations. This paper provides a review of the present understanding of how each of these mechanisms contributes to CM. Original data of the combined effect of stretch and fold and gravitational sedimentation on CM are also presented. Boli of 0.5 microm-diameter particles were inhaled at penetration volumes (V(p)) of 300 and 1200 ml in eight subjects. Inspiration was followed by a 10-s breath hold, during which small flow reversals (FR) were imposed, and expiration. There was no physiologically significant dependence in dispersion and deposition with increasing FR. The results were qualitatively similar to those obtained in a previous study in microgravity in which it was speculated that the phenomenon of stretch and fold occurred during the first breathing cycle without the need of any subsequent FR. PMID- 15890564 TI - Separation and identification of F2-isoprostane regioisomers and diastereomers by novel liquid chromatographic/mass spectrometric methods. AB - Isoprostanes are isomers of prostaglandins that are generated from free radical initiated autoxidation of arachidonic acid. Quantification of F(2)-isoprostanes is regarded as the "gold standard" to assess oxidative stress in various human diseases. There are 32 possible racemic isoprostane isomers that exist as four sets of regioisomers. Each regioisomer is composed of eight diastereomers. We report liquid chromatographic/mass spectrometric methods to separate and identify F(2)-isoprostane stereoisomers. These methods have been applied to the analysis of F(2)-isoprostanes derived from tissues of rats exposed to an oxidative stress and are useful to assess the relative formation of various regioisomers and stereoisomers generated in vitro and in vivo. The delineation of the more abundant isomers formed will allow for studies to examine the biological relevance of selected compounds in vivo. PMID- 15890565 TI - Health and nutrition in Eastern Europe and the former Soviet Union during the decade of transition: a review of the literature. AB - The collapse of the Soviet Union was the most important historical event at the close of the 20th century. The jarring nature of this transition has resulted in large fluctuations in household resources and increased uncertainty in all facets of life for the individuals concerned. Much academic research and popular writing has explored the socioeconomic and political ramifications of bringing these countries into mainstream capitalism. This paper provides a review of the literature examining health outcomes in Eastern Europe and the former Soviet Union during the transition period. The research, which has studied the human face of transition, spans multiple disciplines and it is thus currently difficult for interested researchers to obtain an overview of the basic facts, as well as, the more detailed nuances, concerning developments. This paper highlights what we currently know about health outcomes in transition countries and what we do not know, and suggests future areas of research which may help fill important gaps in our knowledge. PMID- 15890566 TI - Guanidine hydrochloride denaturation of human serum albumin originates by local unfolding of some stable loops in domain III. AB - The effect of guanidine hydrochloride (GnHCl) on the global stability of human serum albumin (HSA) has been studied by fluorescence and circular dichroism spectroscopic measurements. The differential stability of native conformation of three HSA domains were explored by using domain-specific ligands, hemin (domain I), chloroform (domain II), bilirubin (at domain I/domain II interface) and diazepam (domain III). GnHCl induced unfolding transition curves as monitored by probes for secondary and tertiary structures were cooperative but noncoincidental. A strong ANS binding to the protein was observed around 1.8 M GnHCl, suggesting existence of intermediate states in the unfolding pathway of HSA. A gradual decrease (in the GnHCl concentration range 0.0-1.8 M) in the binding of diazepam indicates that domain III is the most labile to GnHCl denaturation. A significant increase in the binding of bilirubin up to 1.4 M GnHCl and decrease thereafter leading to complete abolishment of bilirubin binding at around 2.0 M GnHCl suggest favorable rearrangement and separation of domains I and II at 1.4 and 2.0 M GnHCl concentration, respectively. Above 1.6 M GnHCl, decrease of the binding of hemin, a ligand for domain I, chloroform, which binds in domain II and lone tryptophanyl fluorescence (Trp-214 located in domain II) indicate that at higher concentration of GnHCl domains I and II start unfolding simultaneously but the stability of domain I (7.4 Kcal/mol) is much more than domain II (4.3 Kcal/mol). A pictorial model for the unfolding of HSA domains, consistent with all these results, has been formulated, suggesting that domain III is the most labile followed by domain II while domain I is the most stable. A molten globule like state of domain III around 1.8 M GnHCl has also been identified and characterized. PMID- 15890567 TI - Intraluminal brachytherapy without stenting in intrahepatic papillary cholangiocarcinoma: a case report. AB - A 46-year-old female patient, with mild cholestasis by a large papillary cholangiocarcinoma involving the left hepatic duct, received intraluminal brachytherapy (50 Gy at 1 cm from the source axis) with the aim to relieve biliary obstruction without stent positioning. The patient presented with haemobilia and vegetant lesions in the left main biliary duct, and thus she had a high risk of early stent obstruction. Eighteen months after the treatment the patient presented tumour progression in the controlateral hepatic lobe, but had a patent left hepatic duct, without signs of cholestasis and/or cholangitis. Based on this and other published reports, intraluminal brachytherapy may be tested in a setting different from standard setting with the aim to safely palliate jaundice in patients with intraductal tumour growth in the biliary tract. PMID- 15890568 TI - Maintenance of certification for radiation oncology. AB - Maintenance of Certification (MOC) recognizes that in addition to medical knowledge, several essential elements involved in delivering quality care must be developed and maintained throughout one's career. The MOC process is designed to facilitate and document professional development of American Board of Radiology (ABR) diplomates in the essential elements of quality care in Radiation Oncology and Radiologic Physics. ABR MOC has been developed in accord with guidelines of the American Board of Medical Specialties. All Radiation Oncology certificates issued since 1995 are 10-year, time-limited certificates; diplomates with time limited certificates who wish to maintain specialty certification must complete specific requirements of the American Board of Radiology MOC program. Diplomates with lifelong certificates are not required to participate but are strongly encouraged to do so. Maintenance of Certification is based on documentation of participation in the four components of MOC: (1) professional standing, (2) lifelong learning and self-assessment, (3) cognitive expertise, and (4) performance in practice. Through these components, MOC addresses six competencies medical knowledge, patient care, interpersonal and communication skills, professionalism, practice-based learning and improvement, and systems-based practice. Details of requirements for components 1, 2, and 3 of MOC are outlined along with aspects of the fourth component currently under development. PMID- 15890569 TI - Expression of the epidermal growth factor receptor and Her-2 are predictors of favorable outcome and reduced complete response rates, respectively, in patients with muscle-invading bladder cancers treated by concurrent radiation and cisplatin-based chemotherapy: a report from the Radiation Therapy Oncology Group. AB - PURPOSE: Erb-1 (epidermal growth factor receptor, EGFR) and Erb-2 (Her-2) are two of the best characterized members in the EGFR pathway. In many tumor types, overexpression of these proteins is associated with enhanced malignant potential. Our objective in this study was to investigate the clinical relevance of EGFR and Her-2 expression in bladder cancer cases from four prospective Radiation Therapy Oncology Group (RTOG) bladder preservation trials using cisplatin-containing chemoradiation (RTOG 8802, 8903, 9506, and 9706). METHODS AND MATERIALS: Tumors from 73 cases from patients with muscle-invading T2-T4a bladder cancers had slides interpretable for EGFR staining; 55 cases had slides interpretable for Her 2 staining. Additionally, the respective prognostic values of p53, pRB, and p16 immunostaining were concomitantly examined. Staining and interpretation of staining were done in a blinded manner, without knowledge of clinical outcome. Staining was judged as positive or negative. Subsequently, staining was correlated with clinical outcome. RESULTS: On univariate analysis, EGFR positivity was significantly associated with improved overall survival (p = 0.044); disease-specific survival (DSS) (p = 0.042); and DSS with intact bladder (p = 0.021). There was also a trend for association between EGFR expression and reduced frequency of distant metastasis (p = 0.06). On multivariate analysis adding tumor stage, tumor grade, whether a visibly complete transurethral resection of bladder tumor (TURBT) was done or not, and patient age to the model, EGFR positivity was significantly associated with improved DSS. On univariate analysis, Her-2 positivity was significantly associated with reduced complete response (CR) rates (50% vs. 81%, p = 0.026) after chemoradiation which remained significant on multivariate analysis. The other markers examined in this study were not found to have any prognostic value in this setting. CONCLUSION: Epidermal growth factor receptor expression appears to correlate significantly with improved outcome in bladder cancer, whereas Her-2 expression is significantly associated only with reduced CR rates after chemoradiation. Further investigations are warranted into how EGFR family members regulate response to chemoradiation in bladder cancer and their potential therapeutic implications. PMID- 15890570 TI - Immunohistochemically determined total epidermal growth factor receptor levels not of prognostic value in newly diagnosed glioblastoma multiforme: report from the Radiation Therapy Oncology Group. AB - PURPOSE: The Radiation Therapy Oncology Group (RTOG) performed an analysis of patterns of immunohistochemically detected total epidermal growth factor receptor (EGFR) protein expression levels and their prognostic significance on archival tissue in newly diagnosed glioblastoma multiforme (GBM) patients from prior prospective RTOG clinical trials. METHODS AND MATERIALS: Patients in this study had been treated on previous RTOG GBM trials (RTOG 7401, 7918, 8302, 8409, 9006, 9305, 9602, and 9806). Tissue microarrays were prepared from 155 patients enrolled in these trials. These specimens were stained using a mouse monoclonal antibody specific for the extracellular binding domain of EGFR to detect total EGFR (including both wild-type phosphorylated and wild-type unphosphorylated isoforms with some cross-reactivity with EGFRvIII). The intensity of total EGFR protein expression was measured by computerized quantitative image analysis using the SAMBA 4000 Cell Image Analysis System. The parameters measured were the mean optical densities over the labeled areas and the staining index, which represents the proportion of stained area relative to the mean stain concentration. Both parameters were correlated with the clinical outcome. RESULTS: No differences in either overall or progression-free survival could be demonstrated by the mean optical density class or mean optical density quartile or the staining index of total EGFR immunostaining among the representative RTOG GBM cases. CONCLUSION: Total EGFR protein expression levels, as measured immunohistochemically, do not appear to be of prognostic value in newly diagnosed GBM patients. Given the accumulating clinical evidence of the activity of anti-EGFR agents in GBM and the preclinical data suggesting the important role of downstream mediators as effectors of EGFR signaling, the RTOG is conducting additional investigations into the prognostic value of activation patterns of EGFR signaling, both at the level of the receptor (e.g., EGFRvIII, phospho-EGFR) and at the level of downstream signal transduction pathways (e.g., PI3K, Ras/MAPK pathways). PMID- 15890571 TI - Evaluating changes in tumor volume using magnetic resonance imaging during the course of radiotherapy treatment of high-grade gliomas: Implications for conformal dose-escalation studies. AB - OBJECTIVE: To determine whether changes in tumor volume occur during the course of conformal 3D radiotherapy of high-grade gliomas by use of magnetic resonance imaging (MRI) during treatment and whether these changes had an impact on tumor coverage. METHODS AND MATERIALS: Between December 2000 and January 2004, 21 patients with WHO Grades 3 to 4 supratentorial malignant gliomas treated with 3D conformal radiotherapy (median dose, 70 Gy) were enrolled in a prospective clinical study. All patients underwent T1-weighted contrast-enhancing and T2 weighted and fluid-attenuated inversion recovery (FLAIR) imaging at approximately 1 to 2 weeks before radiotherapy, during radiotherapy (Weeks 1 and 3), and at routine intervals thereafter. All MRI scans were coregistered to the treatment planning CT. Gross tumor volume (GTV Pre-Rx) was defined from a postoperative T1 weighted contrast-enhancing MRI performed 1 to 2 weeks before start of radiotherapy. A second GTV (GTV Week 3) was defined by use of an MRI performed during Week 3 of radiotherapy. A uniform 0.5 cm expansion of the respective GTV, PTV (Pre-Rx), and PTV (Week 3) was applied to the final boost plan. Dose-volume histograms (DVH) were used to analyze any potential adverse changes in tumor coverage based on Week 3 MRI. RESULTS: All MRI scans were reviewed independently by a neuroradiologist (DGH). Two patients were noted to have multifocal disease at presentation and were excluded from analysis. In 19 cases, changes in the GTV based on MRI at Week 3 during radiotherapy were as follows: 2 cases had an objective decrease in GTV (> or =50%); 12 cases revealed a slight decrease in the rim enhancement or changes in cystic appearance of the GTV; 2 cases showed no change in GTV; and 3 cases demonstrated an increase in tumor volume. Both cases with objective decreases in GTV during treatment were Grade 3 tumors. No cases of tumor progression were noted in Grade 3 tumors during treatment. In comparison, three of 12 Grade 4 tumors had tumor progression, based on MRI obtained during Week 3 of radiotherapy. Median increase in GTV (Week 3) was 11.7 cc (range, 9.8 21.3). Retrospective DVH analysis of PTV (Pre-Rx) and PTV (Week 3) demonstrated a decrease in V(95%)(PTV volume receiving 95% of the prescribed dose) in those 3 cases. CONCLUSIONS: Routine MR imaging during radiotherapy may be essential in ensuring tumor coverage if highly conformal radiotherapy techniques such as stereotactic boost and intensity-modulated radiotherapy are used in dose escalation trials that utilize smaller treatment margins. PMID- 15890572 TI - Treatment of pituitary adenomas by fractionated stereotactic radiotherapy: a prospective study of 110 patients. AB - PURPOSE: To optimize and reduce the toxicity of pituitary adenoma irradiation by assessing the feasibility and effectiveness of fractionated stereotactic radiotherapy (FSR). METHODS AND MATERIALS: Between 1990 and 1999, 110 consecutive patients, 47 with a functioning adenoma, were treated according to a strategy of either early surgery and FSR (n = 89) or FSR only (n = 21). Of the 110 patients, 75 had persistent macroscopic tumor and 47 persistent hormonal secretions; 15 were treated in the prophylactic setting. The linear accelerator-delivered dose was 50.4 Gy (5 x 1.8 Gy weekly), with a 2-mm safety margin. RESULTS: After a minimal follow-up of 48 months, only 1 patient had developed progression. Of the 110 patients, 27 (36%) had a complete tumor response, 67 (89.3%) had an objective tumor response, 20 (42%) had a hormonal complete response, and 47 (100%) had a hormonal objective tumor response. The proportion of patients without a complete tumor response, objective tumor response, complete hormonal response, and objective hormonal response was 85.1%, 62%, 83%, and 59.3% at 4 years and 49.3%, 9%, 59.3%, and 10.6% at 8 years, respectively. The sole unfavorable predictive factor was preoperative SSE >20 mm for tumor response (p = 0.01) and growth hormone adenoma for the hormonal response (p <0.001). No late complications, except for pituitary deficiency, were reported, with a probability of requiring hormonal replacement of 28.5% and 35% at 4 and 8 years, respectively. Nonfunctioning status was the sole unfavorable factor (p = 0.0016). CONCLUSIONS: Surgery plus FSR is safe and effective. FSR focused to the target volume seems more suitable than standard radiotherapy, and standard fractionation reduces the risk of optic neuropathy sometimes observed after single-dose radiosurgery. Therefore, FSR allows us to consider combined transrhinoseptal surgery and early radiotherapy, with a curative goal without patient selection. PMID- 15890573 TI - Phase I study of thoracic radiation dose escalation with concurrent chemotherapy for patients with limited small-cell lung cancer: Report of Radiation Therapy Oncology Group (RTOG) protocol 97-12. AB - PURPOSE: The purpose of RTOG 97-12 was to determine the maximum tolerated dose (MTD) of thoracic radiation therapy (RT) with concurrent chemotherapy for patients with limited-stage small-cell lung cancer. PATIENTS AND METHODS: Sixty four patients received four cycles of cisplatin (60 mg/m(2) i.v.) and etoposide (120 mg/m(2) i.v. Days 1-3) (PE), with concurrent thoracic RT starting on Day 1. Thoracic RT was given during the first two cycles with 1.8 Gy/fraction daily to the clinical target volume, followed by thoracic RT to the gross tumor volume b.i.d. for the last 3, 5, 7, 9, or 11 treatment days (total dose 50.4, 54.0, 57.6, 61.2, or 64.8 Gy, respectively). The MTD was based on the dose that produced Grades 3-4 nonhematologic toxicity (mainly esophagitis and pneumonitis) in greater than 50% of patients. RESULTS: After the first 8 patients were enrolled in Arm 1, administration of etoposide was changed from 120 mg/m(2) i.v. on Days 2 and 3 of each cycle to 240 mg/m(2) p.o. for patient convenience as outpatients. Total thoracic RT doses from 50.4 Gy to 61.2 Gy over 5 weeks given with PE were well tolerated. Three of the first 5 patients in the 64.8 Gy arm developed Grade 3 acute esophagitis; the MTD was determined to be 61.2 Gy. Fifty four (87%) of the 62 evaluable patients achieved a complete (68%) or partial (19%) tumor response. The 18-month survival was 25% for patients receiving 50.4 Gy and 82% for those receiving 61.2 Gy. CONCLUSIONS: The MTD for this accelerated thoracic RT regimen with concurrent PE was 61.2 Gy over 5 weeks. PMID- 15890574 TI - Predictors of locoregional recurrence in patients with locally advanced breast cancer treated with neoadjuvant chemotherapy, mastectomy, and radiotherapy. AB - PURPOSE: To identify the clinical and pathologic factors predictive of locoregional recurrence (LRR) after neoadjuvant chemotherapy, mastectomy, and radiotherapy. METHODS AND MATERIALS: We retrospectively reviewed the hospital records of 542 patients treated on six consecutive institutional prospective trials using neoadjuvant chemotherapy and postmastectomy radiotherapy. The clinical stage (American Joint Committee on Cancer, 1988) was Stage II in 17%, Stage IIIA in 30%, Stage IIIB in 43%, and Stage IV (ipsilateral supraclavicular disease) in 10%. All LRRs were considered events, irrespective of the timing to distant metastases. RESULTS: The median follow-up was 70 months. The 5-year and 10-year actuarial LRR rate was 9% and 11%, respectively. The clinical factors associated with LRR included combined clinical stage, clinical T stage, ipsilateral supraclavicular nodal disease, chemotherapy response, physical examination size after chemotherapy, and no tamoxifen use (p < or = 0.04 for all factors). The pathologic predictors of LRR included the number of positive nodes, dissection of <10 nodes, multifocal/multicentric disease, lymphovascular space invasion, extracapsular extension, skin/nipple involvement, and estrogen receptor negative disease (p 10, Gleason > or =7, or MRI evidence of extracapsular extension or seminal vesical invasion) comprised the study cohort. Treatment consisted of hormonal therapy and 4-field 3DCRT using an intrarectal balloon for the initial 15 of 40 treatments. Planning treatment volume dose was 72 Gy (95% normalization). A Mantel-Haenzel Chi-square test compared the distribution of GU, GI, and D symptoms at baseline and at end of treatment (EOT). RESULTS: There was no significant difference between the 2 time points in the proportion of patients with bowel symptoms (p = 0.73), tenesmus (p = 0.27), nocturia (p = 1.00), or GU urgency (p = 0.40). However, there was a significant decrease in GU frequency (70% vs. 50%, p = 0.46) as a result of medical interventions and a significant increase in hemorrhoidal irritation (4% vs. 20%, p = 0.02) and anal cutaneous skin reaction (0% vs. 70%, p < 0.001). By 3 months after EOT compared to baseline, there was no significant difference in the proportion of patients experiencing hemorrhoidal bleeding (4% vs. 8%, p = 0.52), requiring intervention for hemorrhoidal symptoms (7% vs. 5%, p = 0.8), or experiencing persistent anal cutaneous skin reaction (0% vs. 3%, p = 0.31). CONCLUSION: Dose-escalated 3DCRT using an intrarectal balloon for prostate localization and immobilization was well tolerated. Acute GU, GI, and D symptoms resolved with standard dietary or medical interventions by the EOT or shortly thereafter. PMID- 15890581 TI - Long-term androgen deprivation increases Grade 2 and higher late morbidity in prostate cancer patients treated with three-dimensional conformal radiation therapy. AB - PURPOSE: To determine whether the use of androgen deprivation (AD) increases late morbidity when combined with high-dose three-dimensional conformal radiation therapy (3D-CRT). METHODS AND MATERIALS: Between May 1989 and November 1998, 1,204 patients were treated for prostate cancer with 3D-CRT to a median dose of 74 Gy. Patients were evaluated every 3-6 months. No AD was given to 945 patients, whereas 140 and 119 patients, respectively, received short-term AD (STAD; < or =6 months) and long-term AD (LTAD; > 6 months). Radiation morbidity was graded according to the Fox Chase modification of the Late Effects Normal Tissue Task Force late morbidity scale. Covariates in the multivariate analysis (MVA) included age, history of diabetes mellitus, prostate-specific antigen (PSA) level, Gleason score, T category, RT field size, total RT dose, use of rectal shielding, and AD status (no AD vs. STAD vs. LTAD). RESULTS: The only independent predictor for Grade 2 or higher genitourinary (GU) morbidity in the MVA was the use of AD (p = 0.0065). The 5-year risk of Grade 2 or higher GU morbidity was 8% for no AD, 8% for STAD, and 14% for LTAD (p = 0.02). Independent predictors of Grade 2 or higher gastrointestinal (GI) morbidity in the MVA were the use of AD (p = 0.0079), higher total radiation dose (p < 0.0001), the lack of a rectal shield (p = 0.0003), and older age (p = 0.0009). The 5-year actuarial risk of Grade 2 or higher GI morbidity was 17% for no AD vs. 18% for STAD and 26% for LTAD (p = 0.017). CONCLUSIONS: The use of LTAD seems to significantly increase the risk of both GU and GI morbidity for patients treated with 3D-CRT. PMID- 15890582 TI - Prostate gland motion assessed with cine-magnetic resonance imaging (cine-MRI). AB - PURPOSE: To quantify prostate motion during a radiation therapy treatment using cine-magnetic resonance imaging (cine-MRI) for time frames comparable to that expected in an image-guided radiation therapy treatment session (20-30 min). MATERIALS AND METHODS: Six patients undergoing radiation therapy for prostate cancer were imaged on 3 days, over the course of therapy (Weeks 1, 3, and 5). Four hundred images were acquired during the 1-h MRI session in 3 sagittal planes through the prostate at 6-s intervals. Eleven anatomic points of interest (POIs) have been used to characterize prostate/bony pelvis/abdominal wall displacement. Motion traces and standard deviation for each of the 11 POIs have been determined. The probability of displacement over time has also been calculated. RESULTS: Patients were divided into 2 groups according to rectal filling status: full vs. empty rectum. The displacement of POIs (standard deviation) ranged from 0.98 to 1.72 mm for the full-rectum group and from 0.68 to 1.04 mm for the empty rectum group. The low standard deviations in position (2 mm or less) would suggest that these excursions have a low frequency of occurrence. The most sensitive prostate POI to rectal wall motion was the mid-posterior with a standard deviation of 1.72 mm in the full-rectum group vs. 0.79 mm in the empty rectum group (p = 0.0001). This POI has a 10% probability of moving more than 3 mm in a time frame of approximately 1 min if the rectum is full vs. approximately 20 min if the rectum is empty. CONCLUSION: Motion of the prostate and seminal vesicles during a time frame similar to a standard treatment session is reduced compared to that reported in interfraction studies. The most significant predictor for intrafraction prostate motion is the status of rectal filling. A prostate displacement of <3 mm (90%) can be expected for the 20 min after the moment of initial imaging for patients with an empty rectum. This is not the case for patients presenting with full rectum. The determination of appropriate intrafraction margins in radiation therapy to accommodate the time-dependent uncertainty in positional targeting is a topic of ongoing investigations for the on-line image guidance model. PMID- 15890583 TI - Individualized planning target volumes for intrafraction motion during hypofractionated intensity-modulated radiotherapy boost for prostate cancer. AB - PURPOSE: The objective of the study was to access toxicities of delivering a hypofractionated intensity-modulated radiotherapy (IMRT) boost with individualized intrafraction planning target volume (PTV) margins and daily online correction for prostate position. METHODS AND MATERIALS: Phase I involved delivering 42 Gy in 21 fractions using three-dimensional conformal radiotherapy, followed by a Phase II IMRT boost of 30 Gy in 10 fractions. Digital fluoroscopy was used to measure respiratory-induced motion of implanted fiducial markers within the prostate. Electronic portal images were taken of fiducial marker positions before and after each fraction of radiotherapy during the first 9 days of treatment to calculate intrafraction motion. A uniform 10-mm PTV margin was used for the first phase of treatment. PTV margins for Phase II were patient specific and were calculated from the respiratory and intrafraction motion data obtained from Phase I. The IMRT boost was delivered with daily online correction of fiducial marker position. Acute toxicity was measured using National Cancer Institute Common Toxicity Criteria, version 2.0. RESULTS: In 33 patients who had completed treatment, the average PTV margin used during the hypofractionated IMRT boost was 3 mm in the lateral direction, 3 mm in the superior-inferior direction, and 4 mm in the anteroposterior direction. No patients developed acute Grade 3 rectal toxicity. Three patients developed acute Grade 3 urinary frequency and urgency. CONCLUSIONS: PTV margins can be reduced significantly with daily online correction of prostate position. Delivering a hypofractionated boost with this high-precision IMRT technique resulted in acceptable acute toxicity. PMID- 15890584 TI - Impact of geometric uncertainties on evaluation of treatment techniques for prostate cancer. AB - PURPOSE: To assess the impact of patient repositioning and internal organ motion on prostate treatment plans using three-dimensional conformal and intensity modulated radiotherapy. METHODS AND MATERIALS: Four-field, six-field, and simplified intensity-modulated arc therapy plans were generated for 5 prostate cancer patients. The planning target volume was created by adding a 1-cm margin to the clinical target volume. A convolution model was used to estimate the effect of random geometric uncertainties during treatment. Dose statistics, tumor control probabilities, and normal tissue complication probabilities were compared with and without the presence of uncertainty. The impact of systematic uncertainties was also investigated. RESULTS: Compared with the planned treatments, the delivered dose distribution with random geometric uncertainties displayed an increase in the apparent minimal dose to the prostate and seminal vesicles and a decrease in the rectal volume receiving a high dose. This increased the tumor control probabilities and decreased the normal tissue complication probabilities. Changes were seen in the percentage of prostate volume receiving 100% and 95% of the prescribed dose, and the minimal dose and tumor control probabilities for the target volume. In addition, the volume receiving at least 65 Gy, the minimal dose, and normal tissue complication probabilities changed considerably for the rectum. The simplified intensity modulated arc therapy technique was the most sensitive to systematic errors, especially in the anterior-posterior and superior-inferior directions. CONCLUSION: Geometric uncertainties should be considered when evaluating treatment plans. Contrary to the widely held belief, increased conformation of the dose distribution is not always associated with increased sensitivity to random geometric uncertainties if a sufficient planning target volume margin is used. Systematic errors may have a variable effect, depending on the treatment technique used. PMID- 15890585 TI - Erectile function after prostate brachytherapy. AB - PURPOSE: To evaluate erectile function after permanent prostate brachytherapy using a validated patient-administered questionnaire and to determine the effect of multiple clinical, treatment, and dosimetric parameters on penile erectile function. METHODS AND MATERIALS: A total of 226 patients with preimplant erectile function determined by the International Index of Erectile Function (IIEF) questionnaire underwent permanent prostate brachytherapy in two prospective randomized trials between February 2001 and January 2003 for clinical Stage T1c T2c (2002 American Joint Committee on Cancer) prostate cancer. Of the 226 patients, 132 were potent before treatment and, of those, 128 (97%) completed and returned the IIEF questionnaire after brachytherapy. The median follow-up was 29.1 months. Potency was defined as an IIEF score of > or =13. The clinical, treatment, and dosimetric parameters evaluated included patient age; preimplant IIEF score; clinical T stage; pretreatment prostate-specific antigen level; Gleason score; elapsed time after implantation; preimplant nocturnal erections; body mass index; presence of hypertension or diabetes mellitus; tobacco consumption; the volume of the prostate gland receiving 100%, 150%, and 200% of the prescribed dose (V(100/150/200)); the dose delivered to 90% of the prostate gland (D(90)); androgen deprivation therapy; supplemental external beam radiotherapy (EBRT); isotope; prostate volume; planning volume; and radiation dose to the proximal penis. RESULTS: The 3-year actuarial rate of potency preservation was 50.5%. For patients who maintained adequate posttreatment erectile function, the preimplant IIEF score was 29, and in patients with brachytherapy-related ED, the preimplant IIEF score was 25. The median time to the onset of ED was 5.4 months. After brachytherapy, the median IIEF score was 20 in potent patients and 3 in impotent patients. On univariate analysis, the preimplant IIEF score, patient age, presence of nocturnal erections, and dose to the proximal penis predicted for postimplant erectile function. However, in multivariate analysis, only the preimplant IIEF score and the D(50) to the proximal crura were statistically significant predictors of brachytherapy-related erectile function. CONCLUSIONS: Using a patient-administered validated quality-of life instrument, brachytherapy-induced ED occurred in 50% of patients at 3 years. On multivariate analysis, preimplant erectile function and the D(50) to the proximal crura were the best predictors of brachytherapy-related erectile function. Because the proximal penis is the most significant treatment-related predictor of brachytherapy-related ED, techniques to minimize the radiation dose to the proximal penis may result in improved rates of potency preservation. PMID- 15890586 TI - Long-term oncologic results of salvage radical prostatectomy for locally recurrent prostate cancer after radiotherapy. AB - PURPOSE: Salvage radical prostatectomy (RP) may potentially cure patients who have isolated local prostate cancer recurrence after radiotherapy (RT). We report the long-term cancer control associated with salvage RP in a consecutive cohort of patients and identify the variables associated with disease progression and cancer survival. METHODS AND MATERIALS: A total of 100 consecutive patients underwent salvage RP with curative intent for biopsy-confirmed, locally recurrent, prostate cancer after RT. Disease progression after salvage RP was defined as a prostate-specific antigen (PSA) level of > or =0.2 ng/mL or by initiation of androgen deprivation therapy. Cancer-specific mortality was defined as active clinical disease progression despite castration. Cox regression analysis was used to evaluate these endpoints. The median follow-up from RT was 10 years (range, 3-27 years) and from salvage RP was 5 years (range, 1-20 years). RESULTS: Overall, the 5-year progression-free probability was 55% (95% confidence interval, 46-64%), and the median progression-free interval was 6.4 years. The preoperative PSA level was the only significant pretreatment predictor of disease progression in the multivariate analysis (p = 0.01). The 5-year progression-free probability for patients with a preoperative PSA level of <4, 4-10, and >10 ng/mL was 86%, 55%, and 37%, respectively. The 10-year and 15-year cancer-specific mortality after salvage RP was 27% and 40%, respectively. The median time from disease progression to cancer-specific death was 10.3 years (95% confidence interval, 7.6-12.9). After multivariate analysis, the preoperative serum PSA level and seminal vesicle or lymph node status correlated independently with disease progression. CONCLUSIONS: Greater preoperative PSA levels are associated with disease progression and cancer-specific death. Long-term control of locally recurrent prostate cancer after definitive RT is possible when salvage RP is performed early in the course of recurrent disease. PMID- 15890587 TI - Intensity-modulated radiotherapy for adenocarcinoma of the prostate: a point of view. AB - Adenocarcinoma of the prostate (CaP) is treated by surgery or irradiation, or both, with the type of treatment determined largely by local resources and referral patterns. Although the techniques employed by surgeons and radiation oncologists have improved and the morbidities associated with each have declined, for neither are they negligible. Epidemiologic data suggest that between 81% and 85% of men with CaP die of other causes, and a recent survey of untreated men arrived at a similar figure of 83%. Clinical reports, based upon postoperative tumor volume and grade, show that at least 5% of prostatectomies are unnecessary but the extent to which the other 95% benefit from this procedure is unclear. Some sense of these benefits is provided by a randomized, prospective clinical trial that compared prostatectomy with watchful waiting, and found only a 6% gain in overall survival after 8 years. These data call into question the promotion of highly complex and expensive radiation therapy equipment for the treatment of CaP when the prospects for increased life expectancies are at best small and unlikely to be distinguishable from results achieved by surgery, conventional external beam, or radioactive-seed implants. PMID- 15890588 TI - Penile brachytherapy: results for 49 patients. AB - PURPOSE: To report results for 49 men with squamous cell carcinoma (SCC) of the penis treated with primary penile interstitial brachytherapy at one of two institutions: the Ottawa Regional Cancer Center, Ottawa, and the Princess Margaret Hospital, Toronto, Ontario, Canada. METHODS AND MATERIALS: From September 1989 to September 2003, 49 men (mean age, 58 years; range, 22-93 years) had brachytherapy for penile SCC. Fifty-one percent of tumors were T1, 33% T2, and 8% T3; 4% were in situ and 4% Tx. Grade was well differentiated in 31%, moderate in 45%, and poor in 2%; grade was unspecified for 20%. One tumor was verrucous. All tumors in Toronto had pulsed dose rate (PDR) brachytherapy (n = 23), whereas those in Ottawa had either Iridium wire (n = 22) or seeds (n = 4). Four patients had a single plane implant with a plastic tube technique, and all others had a volume implant with predrilled acrylic templates and two or three parallel planes of needles (median, six needles). Mean needle spacing was 13.5 mm (range, 10-18 mm), mean dose rate was 65 cGy/h (range, 33-160 cGy/h), and mean duration was 98.8 h (range, 36-188 h). Dose rates for PDR brachytherapy were 50 61.2 cGy/h, with no correction in total dose, which was 60 Gy in all cases. RESULTS: Median follow-up was 33.4 months (range, 4-140 months). At 5 years, actuarial overall survival was 78.3% and cause-specific survival 90.0%. Four men died of penile cancer, and 6 died of other causes with no evidence of recurrence. The cumulative incidence rate for never having experienced any type of failure at 5 years was 64.4% and for local failure was 85.3%. All 5 patients with local failure were successfully salvaged by surgery; 2 other men required penectomy for necrosis. The soft tissue necrosis rate was 16% and the urethral stenosis rate 12%. Of 8 men with regional failure, 5 were salvaged by lymph node dissection with or without external radiation. All 4 men with distant failure died of disease. Of 49 men, 42 had an intact and tumor-free penis at last follow-up or death. The actuarial penile preservation rate at 5 years was 86.5%. CONCLUSIONS: Brachytherapy is an effective treatment for T1, T2, and selected T3 SCC of the penis. Close follow-up is mandatory because local failures and many regional failures can be salvaged by surgery. PMID- 15890589 TI - Pilot study in the treatment of endometrial carcinoma with 3D image-based high dose-rate brachytherapy using modified Heyman packing: clinical experience and dose-volume histogram analysis. AB - PURPOSE: The aim of this study was to evaluate dose distribution within uterus (clinical target volume [CTV]) and tumor (gross tumor volume [GTV]) and the resulting clinical outcome based on systematic three-dimensional treatment planning with dose-volume adaptation. Dose-volume assessment and adaptation in organs at risk and its impact on side effects were investigated in parallel. METHODS AND MATERIALS: Sixteen patients with either locally confined endometrial carcinoma (n = 15) or adenocarcinoma of uterus and ovaries after bilateral salpingo-oophorectomy (n = 1) were included. Heyman packing was performed with mean 11 Norman-Simon applicators (3-18). Three-dimensional treatment planning based on computed tomography (n = 29) or magnetic resonance imaging (n = 18) was done in all patients with contouring of CTV, GTV, and organs at risk. Dose-volume adaptation was achieved by dwell location and time variation (intensity modulation). Twelve patients treated with curative intent received five to seven fractions of high-dose-rate brachytherapy (7 Gy per fraction) corresponding to a total dose of 60 Gy (2 Gy per fraction and alpha/beta of 10 Gy) to the CTV. Four patients had additional external beam radiotherapy (range, 10-40 Gy). One patient had salvage brachytherapy and 3 patients were treated with palliative intent. A dose-volume histogram analysis was performed in all patients. On average, 68% of the CTV and 92% of the GTV were encompassed by the 60 Gy reference volume. Median minimum dose to 90% of CTV and GTV (D90) was 35.3 Gy and 74 Gy, respectively. RESULTS: All patients treated with curative intent had complete remission (12/12). After a median follow-up of 47 months, 5 patients are alive without tumor. Seven patients died without tumor from intercurrent disease after median 22 months. The patient with salvage treatment had a second local recurrence after 27 months and died of endometrial carcinoma after 57 months. In patients treated with palliative intent, symptom relief was achieved. No severe acute and late side effects (Grade 3/4) were observed. CONCLUSIONS: Sectional image-based three dimensional treatment planning on computed tomography and magnetic resonance imaging is feasible in definitive brachytherapy of endometrial carcinoma and enables by the use of dwell time and location adaptation a sufficient coverage of GTV and major parts of CTV. Local control in this limited number of patients is excellent and rate of side effects minimal. PMID- 15890590 TI - Anal canal carcinoma: early-stage tumors < or =10 mm (T1 or Tis): therapeutic options and original pattern of local failure after radiotherapy. AB - PURPOSE: To investigate the clinical history, management, and pattern of recurrence of very early-stage anal canal cancer in a French retrospective survey. METHODS: The study group consisted of 69 patients with Stage Tis and T1 anal canal carcinoma < or =1 cm treated between 1990 and 2000 (12 were in situ, 57 invasive, 66 Stage N0, and 3 Stage N1). The median patient age was 67 years (range, 27-83 years). Of the 69 patients, 66 received radiotherapy (RT) and 3 with in situ disease were treated by local excision alone without RT. Twenty-six patients underwent local excision before RT (12 with negative and 14 with positive surgical margins). Of the 66 patients who underwent RT, 8 underwent brachytherapy alone (median dose, 55 Gy), 38 underwent external beam RT (median dose, 45 Gy) plus a brachytherapy boost (median boost dose, 20 Gy), and 20 underwent external beam RT alone (median dose, 55 Gy). RESULTS: Of the 69 patients, 68 had initial local control. Of the 66 patients treated by RT, 6 developed local recurrence at a median interval of 50 months (range, 13-78 months). Four patients developed local failure outside the initial tumor bed. Of the 3 patients with Tis treated by excision alone, 1 developed local recurrence. No relation was found among prior excision, dose, and local failure. The 5-year overall survival, colostomy-free survival, and disease-free survival rate was 94%, 85%, and 89%, respectively. The rate of late complications (Grade 1-3) was 28% and was 14% for those who received doses <60 Gy and 37% for those who received doses of > or =60 Gy (p = 0.04). CONCLUSION: Most recurrences occurred after a long disease-free interval after treatment and often outside the initial tumor site. These small anal cancers could be treated by RT using a small volume and moderate dose (40-50 Gy for subclinical lesions and 50-60 Gy for T1). PMID- 15890591 TI - Phase II multicenter randomized study of amifostine for prevention of acute radiation rectal toxicity: topical intrarectal versus subcutaneous application. AB - PURPOSE: To investigate the cytoprotective effect of subcutaneous vs. intrarectal administration of amifostine against acute radiation toxicity. METHODS AND MATERIALS: Patients were randomized to receive amifostine either intrarectally (Group A, n = 27) or a 500-mg flat dose subcutaneously (Group B, n = 26) before irradiation. Therapy was delivered using a four-field technique with three dimensional conformal planning. In Group A, 1,500 mg of amifostine was administered intrarectally as an aqueous solution in 40 mL of enema. Two different toxicity scales were used: the European Organization for Research and Treatment of Cancer/Radiation Therapy Oncology Group (RTOG) rectal and urologic toxicity criteria and the Subjective-RectoSigmoid scale based on the endoscopic terminology of the World Organization for Digestive Endoscopy. Objective measurements with rectosigmoidoscopy were performed at baseline and 1-2 days after radiotherapy completion. The area under the curve for the time course of mucositis (RTOG criteria) during irradiation represented the mucositis index. RESULTS: Intrarectal amifostine was feasible and well tolerated without any systemic or local side effects. According to the RTOG toxicity scale, Group A had superior results with a significantly lower incidence of Grades I-II rectal radiation morbidity (11% vs. 42%, p = 0.04) but inferior results concerning urinary toxicity (48% vs. 15%, p = 0.03). The mean rectal mucositis index and Subjective-RectoSigmoid score were significantly lower in Group A (0.44 vs. 2.45 [p = 0.015] and 3.9 vs. 6.0 [p = 0.01], respectively), and the mean urinary mucositis index was lower in Group B (2.39 vs. 0.34, p < 0.028). CONCLUSIONS: Intrarectal administration of amifostine (1,500 mg) seemed to have a cytoprotective efficacy in acute radiation rectal mucositis but was inferior to subcutaneous administration in terms of urinary toxicity. Additional randomized studies are needed for definitive decisions concerning the cytoprotection of pelvic irradiated areas. PMID- 15890592 TI - Proton radiation for treatment of cancer of the oropharynx: early experience at Loma Linda University Medical Center using a concomitant boost technique. AB - PURPOSE: To assess accelerated fractionation using photon and proton radiation to improve local control and reduce complications in treating locally advanced oropharyngeal cancer. METHODS AND MATERIALS: Twenty-nine patients with localized Stage II-IV oropharyngeal cancer received accelerated photon and proton radiation, 75.9 GyE in 45 fractions/5.5 weeks, to the primary disease, involved lymph nodes, and potential areas of subclinical spread. Follow-up ranged from 2 to 96 months. RESULTS: Five-year actuarial control for local disease was 88%, and for neck node disease, 96%; yielding a 84% locoregional control rate at 5 years. Four patients developed distant metastases. The 5-year actuarial locoregional control rate was 84%. The actuarial 2-year disease-free survival rate was 81%; at 5 years, it was 65%. All patients completed the prescribed treatment; though aggressive nutritional and anesthetic support was necessary. Late Grade 3 toxicity was seen in 3 patients. CONCLUSIONS: Protons used as a concomitant boost with photons effectively delivered an accelerated time-dose schedule to the cancer with a more tolerable schedule to surrounding normal tissues. Preliminary results reveal increased locoregional control without increased toxicity. Future studies must evaluate the optimum time-dose schedule. PMID- 15890593 TI - Nasopharyngeal carcinoma staging by (18)F-fluorodeoxyglucose positron emission tomography. AB - PURPOSE: Nasopharyngeal carcinoma (NPC) has a high rate of neck lymph node and/or distant metastasis. We evaluated the value of (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) in staging NPC, especially in the detection of distant metastasis. METHODS AND MATERIALS: A total of 95 patients, including 85 with primary and 10 with recurrent, NPC were enrolled. Dual-phase FDG-PET was used, in addition to the conventional workup. Eighty-one patients without distant metastases underwent repeat studies 3-4 months after initial radical treatment. RESULTS: Of 14 patients with distant metastases, all had lesions detected by FDG PET, and the conventional workup detected the metastases in only 4. Two patients had false-positive MRI findings for neck node metastasis, but the FDG-PET findings were accurate. Four patients without distant metastases on their initial workup were found to have new lesions on FDG-PET 3-4 months after initial treatment. Patients with advanced node disease had a significantly greater incidence of distant metastases on FDG-PET, especially for N3 disease. Of the 95 patients, the FDG-PET results for distant metastasis were true positive in 14 patients, false positive in 8, and true negative in 73. None of our patients had a false-negative result. For a patient base, the sensitivity and specificity of FDG-PET for distant metastasis was 100% and 90.1% (95% confidence interval 81.5 95.6%), respectively, in this study. The accuracy was 91.6% (95% confidence interval 84.1-96.3%), the positive predictive value was 63.6 (95% confidence interval 40.7-82.8%), and the negative predictive value was 100%. CONCLUSION: FDG PET stages N and M disease of NPC more accurately and sensitively than does the conventional workup. Patients with advanced node disease, particularly N3 disease, would benefit the most from FDG-PET. PMID- 15890594 TI - Neoadjuvant chemotherapy followed by concurrent chemo-radiation therapy in locally advanced nasopharyngeal carcinoma. AB - PURPOSE: To evaluate the efficacy and outcomes of neoadjuvant cisplatinum and epirubicin chemotherapy followed by concurrent cisplatinum chemotherapy with radiotherapy in patients with locally advanced nasopharyngeal carcinoma. METHODS AND MATERIALS: One hundred ten patients (80 male, 30 female) with locally advanced nasopharyngeal carcinoma, staged according to the 1997 International Union Against Cancer/American Joint Committee on Cancer classification system as IIB (n = 9), III (n = 20), IVA (n = 32), and IVB (n = 49), World Health Organization types II (n = 25) and III (n = 85), were included in this protocol between January 1998 and July 2000 at King Faisal Specialist Hospital and Research Centre. Patients underwent two cycles of induction chemotherapy with cisplatinum 100 mg/m(2) and epirubicin 70 mg/m(2) on Days 1 and 21, followed by a radical course of radiotherapy (6,600 cGy in 6.5 weeks, 200 cGy/fraction) starting on Day 42, with three cycles of concurrent cisplatinum 25 mg/m(2) for 4 days on Days 42, 63, and 84. RESULTS: Of 110 patients included in this study, intracranial extension was present in 32 (29%), and nodal stage was N3 in 49 (45%). Complete remission and partial remission were achieved in 87 patients (79%) and 23 patients (21%), respectively. At a median follow-up for surviving patients of 37 months (22-55 months), 49 of 110 patients (44%) had failed treatment: 12 with local, 9 with regional nodes, 4 locoregional, 5 locoregional plus distant areas, and 19 with distant metastases. At the time of writing, 34 patients had died; all deaths were related to the patients' cancer except for 1 patient with treatment-related toxicity. Three-year actuarial overall survival, relapse-free survival, locoregional control, and distant metastasis-free survival rates were 89%, 78%, 88%, and 89% for patients with stage IIB; 71%, 70%, 89%, and 74% for stage III; 68%, 49%, 61%, and 77% for stage IVA; and 70%, 45%, 60%, and 69% for stage IVB, respectively. One patient received only one induction cycle; all others received two cycles; however, 9 of them required 20% reduction in the second cycle dose. Ninety patients (82%) completed two or more concurrent cycles of cisplatinum. Rates of Grade 3 and 4 reactions after induction chemotherapy were as follows: anemia 1% and 0%, leukopenia 8% and 4%, nausea 27% and 0%, vomiting 25% and 0%, and infection 4% and 4%, respectively. Acute Grade 3 and 4 reactions were also observed during chemoradiotherapy: anemia 1% and 0%, leukopenia 31% and 4%, nausea 35% and 0%, vomiting 26% and 2%, infection in 4% and 2%, mucositis in 49% and 0%, and skin reaction in 39% and 0%, respectively. CONCLUSIONS: Neoadjuvant chemotherapy followed by concurrent chemoradiotherapy is a safe and effective method of treatment for locally advanced nasopharyngeal carcinoma. Further investigations in prospective studies are required to evaluate this regimen. PMID- 15890595 TI - Comparison of 1 x 8 Gy and 10 x 3 Gy for functional outcome in patients with metastatic spinal cord compression. AB - PURPOSE: Since life expectancy is markedly reduced in patients with metastatic spinal cord compression (MSCC), a short and effective radiation schedule is desired. This study investigates a reduction of the overall treatment time to only one day by comparing 1 x 8 Gy to the multi-fractionated 10 x 3 Gy for functional outcome. METHODS AND MATERIALS: Data of 204 patients, treated for MSCC with either 1 x 8 Gy (n = 96) or 10 x 3 Gy (n = 108), were analyzed retrospectively. Motor function and ambulatory status were evaluated before and up to 24 weeks after RT. A multivariate analysis (nominal regression) was performed including radiation schedule, performance status, age, irradiated vertebra, and relevant prognostic factors (histology, ambulatory status, time of developing motor deficits). Improvement of motor deficits was selected as basic category and compared with no change and deterioration. RESULTS: Univariate analysis showed no significant difference between the schedules for post treatment motor function and ambulatory rates. Multivariate analysis demonstrated a significant effect on functional outcome for the prognostic factors, but not for the radiation schedule (p = 0.853 for no change, p = 0.237 for deterioration). CONCLUSIONS: Our data suggest the two fractionation schedules to be comparably effective for functional outcome. Thus, 1 x 8 Gy should be considered for patients with a poor survival prognosis. PMID- 15890596 TI - Radiation oncology career decision variables for graduating trainees seeking positions in 2003-2004. AB - PURPOSE: Radiation oncology trainees must consider an array of variables when deciding upon an academic or private practice career path. This prospective evaluation of the 2004 graduating radiation oncology trainees, evaluates such variables and provides additional descriptive data. METHODS: A survey that included 15 questions (one subjective, eleven categorical, and 3 continuous variables) was mailed to the 144 graduating radiation oncology trainees in United States programs in January of 2004. Questions were designed to gather information regarding factors that may have influenced career path choices. The responses were anonymous, and no identifying information was sought. Survey data were collated and analyzed for differences in both categorical and continuous variables as they related to choice of academic or private practice career path. RESULTS: Sixty seven (47%) of the surveys were returned. Forty-five percent of respondents indicated pursuit of an academic career. All respondents participated in research during training with 73% participating in research publication authorship. Post graduate year-3 was the median in which career path was chosen, and 20% thought that a fellowship position was "perhaps" necessary to secure an academic position. Thirty percent of the respondents revealed that the timing of the American Board of Radiology examination influenced their career path decision. Eighteen variables were offered as possibly influencing career path choice within the survey, and the top five identified by those seeking an academic path were: (1) colleagues, (2) clinical research, (3) teaching, (4) geography, (5) and support staff. For those seeking private practice, the top choices were: (1) lifestyle, (2) practice environment, (3) patient care, (4) geography, (5) colleagues. Female gender (p = 0.064), oral meeting presentation (p = 0.053), and international meeting presentation (p = 0.066) were the variables most significantly associated with pursuing an academic career path. The following variables were ranked significantly differently in hierarchy (p < 0.05) by those seeking an academic versus private practice path with respect to having influence on the career decision: lifestyle, income, case-mix, autonomy, ability to sub-specialize, basic research, clinical research, teaching, patient care, board structure, practice environment, and mentoring. CONCLUSION: These data offer descriptive information regarding variables that lead to radiation oncology trainee career path decisions. Such information may be of use in modification of training programs to meet future personnel and programmatic needs within the specialty. PMID- 15890597 TI - 2003 survey of Canadian radiation oncology residents. AB - PURPOSE: Radiation oncology's popularity as a career in Canada has surged in the past 5 years. Consequently, resident numbers in Canadian radiation oncology residencies are at all-time highs. This study aimed to survey Canadian radiation oncology residents about their opinions of their specialty and training experiences. METHODS AND MATERIALS: Residents of Canadian radiation oncology residencies that enroll trainees through the Canadian Resident Matching Service were identified from a national database. Residents were mailed an anonymous survey. RESULTS: Eight of 101 (7.9%) potential respondents were foreign funded. Fifty-two of 101 (51.5%) residents responded. A strong record of graduating its residents was the most important factor residents considered when choosing programs. Satisfaction with their program was expressed by 92.3% of respondents, and 94.3% expressed satisfaction with their specialty. Respondents planning to practice in Canada totaled 80.8%, and 76.9% plan to have academic careers. Respondents identified job availability and receiving adequate teaching from preceptors during residency as their most important concerns. CONCLUSIONS: Though most respondents are satisfied with their programs and specialty, job availability and adequate teaching are concerns. In the future, limited time and resources and the continued popularity of radiation oncology as a career will magnify the challenge of training competent radiation oncologists in Canada. PMID- 15890598 TI - Apoptosis and proliferation of oligodendrocyte progenitor cells in the irradiated rodent spinal cord. AB - PURPOSE: Oligodendrocytes undergo early apoptosis after irradiation. The aim of this study was to determine the relationship between oligodendroglial apoptosis and proliferation of oligodendrocyte progenitor cells (OPC) in the irradiated central nervous system. METHODS AND MATERIALS: Adult rats and p53 transgenic mice were given single doses of 2 Gy, 8 Gy, or 22 Gy to the cervical spinal cord. Apoptosis was assessed using TUNEL (Tdt-mediated dUTP terminal nick-end labeling) staining or by examining nuclear morphology. Oligodendrocyte progenitor cells were identified with an NG2 antibody or by in situ hybridization for platelet derived growth factor receptor alpha. Proliferation of OPC was assessed by in vivo bromodeoxyuridine (BrdU) labeling and subsequent immunohistochemistry. Because radiation-induced apoptosis of oligodendroglial cells is p53 dependent, p53 transgenic mice were used to study the relationship between apoptosis and cell proliferation. RESULTS: Oligodendrocyte progenitor cells underwent apoptosis within 24 h of irradiation in the rat. That did not result in a change in OPC density at 24 h. Oligodendrocyte progenitor cell density was significantly reduced by 2-4 weeks, but showed recovery by 6 weeks after irradiation. An increase in BrdU-labeled cells was observed at 2 weeks after 8 Gy or 22 Gy, and proliferating cells in the rat spinal cord were immunoreactive for NG2. The mouse spinal cord showed a similar early cell proliferation after irradiation. No difference was observed in the proliferation response in the spinal cord of p53 /- mice compared with wild type animals. CONCLUSIONS: Oligodendroglial cells undergo early apoptosis and OPC undergo early proliferation after ionizing radiation. However, apoptosis is not likely to be the trigger for early proliferation of OPC in the irradiated central nervous system. PMID- 15890599 TI - Dependence of FDG uptake on tumor microenvironment. AB - PURPOSE: To investigate the factors affecting the (18)F-fluorodeoxyglucose ((18)F FDG) uptake in tumors at a microscopic level, by correlating it with tumor hypoxia, cellular proliferation, and blood perfusion. METHODS AND MATERIALS: Nude mice bearing Dunning prostate tumors (R3327-AT) were injected with (18)F-FDG and pimonidazole, bromodeoxyuridine, and, 1 min before sacrifice, with Hoechst 33342. Selected tumor sections were imaged by phosphor plate autoradiography, while adjacent sections were used to obtain the images of the spatial distribution of Hoechst 33342, pimonidazole, and bromodeoxyuridine. The images were co-registered and analyzed on a pixel-by-pixel basis. RESULTS: Statistical analysis of the data obtained from these tumors demonstrated that (18)F-FDG uptake was positively correlated with pimonidazole staining intensity in each data set studied. Correlation of FDG uptake with bromodeoxyuridine staining intensity was always negative. In addition, FDG uptake was always negatively correlated with the staining intensity of Hoechst 33342. CONCLUSIONS: For the Dunning prostate tumors studied, FDG uptake was always positively correlated with hypoxia and negatively correlated with both cellular proliferation and blood flow. Therefore, for the tumor model studied, higher FDG uptake is indicative of tumor hypoxia, but neither blood flow nor cellular proliferation. PMID- 15890600 TI - Benefit of respiration-gated stereotactic radiotherapy for stage I lung cancer: an analysis of 4DCT datasets. AB - PURPOSE: High local control rates have been reported with stereotactic radiotherapy (SRT) for Stage I non-small-cell lung cancer. Because high-dose fractions are used, reduction in treatment portals will reduce the risk of toxicity to adjacent structures. Respiratory gating can allow reduced field sizes and planning four-dimensional computed tomography scans were retrospectively analyzed to study the benefits for gated SRT and identify patients who derive significant benefit from this approach. METHODS AND MATERIALS: A total of 31 consecutive patients underwent a four-dimensional computed tomography scan, in which three-dimensional computed tomography datasets for 10 phase bins of the respiratory cycle were acquired during free breathing. For a total of 34 tumors, the three planning target volumes (PTVs) were analyzed, namely (1) PTV(10bins), derived from an internal target volume (ITV) that incorporated all observed mobility (ITV(10bins)), with the addition of a 3-mm isotropic setup margin; (2) PTV(gating), derived from an ITV generated from mobility observed in three consecutive phases ("bins") during tidal-expiration, plus addition of a 3-mm isotropic margin; and (3) PTV(10 mm), derived from the addition of a 10-mm isotropic margin to the most central gross tumor volumes in the three bins selected for gating. RESULTS: The PTV(10bins) and PTV(gating) were, on average, 48.2% and 33.3% of the PTV(10 mm), and respective mean volumes of normal tissue (outside the PTV) receiving the prescribed doses were 57.1% and 39.1%, respectively, of that of PTV(10 mm). A significant correlation was seen between the extent of tumor mobility (i.e., a three-dimensional mobility vector of at least 1 cm) and reduction in normal tissue irradiation achieved with gating. The ratio of the intersecting and the encompassing volumes of GTVs at extreme phases of tidal respiration predicted for the benefits of gated respiration. CONCLUSION: The use of "standard population-based" margins for SRT leads to unnecessary normal tissue irradiation. The risk of toxicity is further reduced if respiration gated radiotherapy is used to treat mobile tumors. These findings suggest that gated SRT will be of clinical relevance in selected patients with mobile tumors. PMID- 15890601 TI - Dose heterogeneity in the target volume and intensity-modulated radiotherapy to escalate the dose in the treatment of non-small-cell lung cancer. AB - PURPOSE: To quantify the dose escalation achievable in the treatment of non-small cell lung cancer (NSCLC) by allowing dose heterogeneity in the target volume or using intensity-modulated radiotherapy (IMRT), or both. METHODS AND MATERIALS: Computed tomography data and contours of 10 NSCLC patients with limited movements of the tumor and representing a broad spectrum of clinical cases were selected for this study. Four irradiation techniques were compared: two conformal (CRT) and two IMRT techniques, either prescribing a homogeneous dose in the planning target volume (PTV) (CRT(hom) and IMRT(hom)) or allowing dose heterogeneity (CRT(inhom) and IMRT(inhom)). The dose heterogeneity was allowed only toward high doses, i.e., the minimum dose in the target for CRT(inhom) and IMRT(inhom) could not be lower than for the corresponding homogeneous plan. The dose in the PTV was escalated (fraction size of 2.25 Gy) until either an organ at risk reached the maximum allowed dose or the mean PTV dose reached a maximum level set at 101.25 Gy. RESULTS: When small and convex tumors were irradiated, CRT(hom) could achieve the maximum dose of 101.25 Gy, whereas for bigger and/or concave PTVs the dose level achievable with CRT(hom) was significantly lower, in 1 case even below 60 Gy. The CRT(inhom) allowed on average a 6% dose escalation with respect to CRT(hom). The IMRT(hom) achieved in all except 1 case a mean PTV dose of at least 75 Gy. The gain in mean PTV dose of IMRT(hom) with respect to CRT(hom) ranged from 7.7 to 14.8 Gy and the IMRT(hom) plans were always more conformal than the corresponding CRT(hom) plans. The IMRT(inhom) provided an additional advantage over IMRT(hom) of at least 5 Gy. For all CRT plans the achievable dose was determined by the lung dose threshold, whereas for more than half of the IMRT plans the esophagus was the dose-limiting organ. The IMRT plans were deliverable with 10-12 segments per beam and did not produce an increase of lung volume irradiated at low doses (<20 Gy). CONCLUSIONS: The dose in NSCLC treatments can be escalated by loosening the constraints on maximum dose in the target volume or using IMRT, or both. For large and concave tumors, an average dose escalation of 6% and 17% was possible when dose heterogeneity and IMRT were applied alone. When they were combined, the average dose increase was as high as 35%. Intensity modulated RT delivered in a static mode can produce homogeneous dose distributions in the target and does not lead to an increase of lung volume receiving (very) low doses, even down to 5 Gy. PMID- 15890602 TI - Benefit of using biologic parameters (EUD and NTCP) in IMRT optimization for treatment of intrahepatic tumors. AB - PURPOSE: To investigate whether intensity-modulated radiotherapy (IMRT), optimized using the generalized equivalent uniform dose (gEUD) and normal tissue complication probability (NTCP) models, can increase the safe dose to intrahepatic tumors compared with three-dimensional conformal RT (3D-CRT). A secondary objective was to investigate the optimal beam arrangement for liver IMRT plans. METHODS AND MATERIALS: Planning CT data of 15 patients with intrahepatic tumors, previously treated with 3D-CRT, were used as input. The dose delivered using 3D-CRT had been limited either by tolerance of adjacent organs, which were close to, or overlapped with, the planning target volume (PTV; overlap cases, n = 8), or liver toxicity (nonoverlap, n = 7). IMRT plans were created using the gEUD to maximize the dose across the PTV and the NTCP to maintain the organ-at-risk toxicity to that of the conformal plan. Increased heterogeneity was allowed across the PTV in overlap cases, without compromising the minimal PTV dose of the conformal plan and restricting the maximal dose to within 110% of the mean. Three different beam arrangements were used for each case: seven-field equidistant axial, six-field noncoplanar (predominantly right-sided beams), and a reproduction of the conformal gantry angles. gEUDs were also computed and used for evaluation of the plans (regardless of planning technique) to reflect the response of both high- and low-grade tumors. The IMRT plan that allowed the greatest gEUD across the PTV was used in the comparison with the 3D-CRT plan. RESULTS: The use of IMRT significantly increased the maximal gEUD achievable across the PTV compared with the 3D-CRT plans. This was the case for the assumptions of both high- and low-grade tumors, irrespective of the tumor position within the liver. The mean gEUD increase was 11 Gy (high grade) and 18.0 Gy (low grade) for overlap cases (p = 0.001 and p = 0.003, respectively) and 10 Gy for nonoverlap cases (p = 0.020). When comparing the IMRT beam arrangements, gEUDs were considered equivalent if they differed by less than one fraction (1.5 Gy). In overlap cases (n = 8), an equivalent "best" gEUD value was obtained in 3, 5, and 7 cases for the original conformal angle, seven-field axial, and six-field noncoplanar plan, respectively. The corresponding results were 5, 2, and 3 in the cases without an overlap (n = 7). CONCLUSION: We have successfully used mathematical/biologic models directly as cost functions within the optimizing process to produce IMRT plans that maximize the gEUD while maintaining compliance with a well-defined protocol for the treatment of intrahepatic cancer. For both PTV-organ-at-risk overlap and nonoverlap situations, IMRT has the capacity to improve the maximal dose achievable across the PTV, expressed in terms of the gEUD. The use of multiple noncoplanar beams appears to confer an advantage over fewer beams in cases with PTV-organ-at-risk overlap. When liver toxicity is the dose-limiting factor, high gEUD values are obtained most frequently when the field arrangement is chosen to provide the shortest possible transhepatic path length. PMID- 15890603 TI - A dosimetric study of Leipzig applicators. AB - PURPOSE: To obtain the absolute dose-rate distribution in liquid water for all six cup-shaped Leipzig applicators by means of an experimentally validated Monte Carlo (MC) code. These six applicators were used in high-dose-rate (HDR) afterloaders with the "classic" and v2 (192)Ir sources. The applicators have an inner diameter of 1, 2, and 3 cm, with the source traveling parallel or perpendicular to the contact surface. METHODS AND MATERIALS: The MC GEANT4 code was used to obtain the dose-rate distribution in liquid water for the six applicators and the two HDR source models. To normalize the applicator output factors, a MC simulation for the "classic" and v2 sources in air was performed to estimate the air-kerma strength. To validate this specific application and to guarantee that realistic source-applicator geometry was considered, an experimental verification procedure was implemented in this study, in accordance with the TG43U1 recommendations. Thermolumniscent dosimeter chips and a parallel plate ionization chamber in a polymethyl methacrylate (PMMA) phantom were used to verify the MC results for the six applicators in a microSelectronHDR afterloader with the "classic" source. Dose-rate distributions dependence on phantom size has been evaluated using two different phantom sizes. RESULTS: Percentage depth dose and off-axis profiles were obtained normalized at a depth of 3 mm along the central axis for both phantom sizes. A table of output factors, normalized to 1 U of source kerma strength at this depth, is presented. The dose measured in the PMMA phantom agrees within experimental uncertainties with the dose obtained by the MC GEANT4 code calculations. The phantom size influence on dose-rate distributions becomes significant at depths greater than 5 cm. CONCLUSIONS: MC detailed simulation was performed for the Nucletron Leipzig HDR applicators. The matrix data obtained, with a grid separation of 0.5 mm, can be used to build a dataset in a convenient format to model these distributions for routine use with a brachytherapy treatment planning system. PMID- 15890604 TI - (144)Ce as a potential candidate for interstitial and intravascular brachytherapy. AB - PURPOSE: To investigate the suitability of (144)Ce for both interstitial and intravascular brachytherapy applications. METHODS AND MATERIALS: Monte Carlo calculations of radial dose rate distributions in water were performed for (144)Ce in a spring-shaped source and compared with two commonly used interstitial and intravascular sources, (192)Ir and (32)P. The numeric simulations were checked experimentally with a calibrated ionization chamber in a water phantom. Other source characteristics, such as half-life and specific activity, were also compared. RESULTS: For interstitial brachytherapy, (144)Ce presents dosimetric advantages over (192)Ir in terms of higher dose rate at shorter distances and lower irradiation of organs outside the tumor. The source size and shape reduce the anisotropy and the number of dwell positions necessary. The longer half-life of (144)Ce might also be advantageous over (192)Ir. For intravascular brachytherapy, (144)Ce permits the treatment of larger arteries as compared with (32)P, compensates centering errors more effectively, and has a more suitable half-life. The experimental validation showed good agreement (within 10%) with the Monte Carlo simulated dose rate distributions. CONCLUSIONS: There are certain potential advantages of using (144)Ce as a source for both interstitial and intravascular brachytherapy. The basis for this finding is provided by the Monte Carlo radial dose rate comparisons with (192)Ir and (32)P, as well as by such characteristics as half-life and specific activity. PMID- 15890605 TI - Narrow band deformable registration of prostate magnetic resonance imaging, magnetic resonance spectroscopic imaging, and computed tomography studies. AB - PURPOSE: Endorectal (ER) coil-based magnetic resonance imaging (MRI) and magnetic resonance spectroscopic imaging (MRSI) is often used to obtain anatomic and metabolic images of the prostate and to accurately identify and assess the intraprostatic lesions. Recent advancements in high-field (3 Tesla or above) MR techniques affords significantly enhanced signal-to-noise ratio and makes it possible to obtain high-quality MRI data. In reality, the use of rigid or inflatable endorectal probes deforms the shape of the prostate gland, and the images so obtained are not directly usable in radiation therapy planning. The purpose of this work is to apply a narrow band deformable registration model to faithfully map the acquired information from the ER-based MRI/MRSI onto treatment planning computed tomography (CT) images. METHODS AND MATERIALS: A narrow band registration, which is a hybrid method combining the advantages of pixel-based and distance-based registration techniques, was used to directly register ER based MRI/MRSI with CT. The normalized correlation between the two input images for registration was used as the metric, and the calculation was restricted to those points contained in the narrow bands around the user-delineated structures. The narrow band method is inherently efficient because of the use of a priori information of the meaningful contour data. The registration was performed in two steps. First, the two input images were grossly aligned using a rigid registration. The detailed mapping was then modeled by free form deformations based on B-spline. The limited memory Broyden-Fletcher-Goldfarb-Shanno algorithm (L-BFGS), which is known for its superior performance in dealing with high dimensionality problems, was implemented to optimize the metric function. The convergence behavior of the algorithm was studied by self-registering an MR image with 100 randomly initiated relative positions. To evaluate the performance of the algorithm, an MR image was intentionally distorted, and an attempt was then made to register the distorted image with the original one. The ability of the algorithm to recover the original image was assessed using a checkerboard graph. The mapping of ER-based MRI onto treatment planning CT images was carried out for two clinical cases, and the performance of the registration was evaluated. RESULTS: A narrow band deformable image registration algorithm has been implemented for direct registration of ER-based prostate MRI/MRSI and CT studies. The convergence of the algorithm was confirmed by starting the registration experiment from more than 100 different initial conditions. It was shown that the technique can restore an MR image from intentionally introduced deformations with an accuracy of approximately 2 mm. Application of the technique to two clinical prostate MRI/CT registrations indicated that it is capable of producing clinically sensible mapping. The whole registration procedure for a complete three-dimensional study (containing 256 x 256 x 64 voxels) took less than 15 min on a standard personal computer, and the convergence was usually achieved in fewer than 100 iterations. CONCLUSIONS: A deformable image registration procedure suitable for mapping ER-based MRI data onto planning CT images was presented. Both hypothetical tests and patient studies have indicated that the registration is reliable and provides a valuable tool to integrate the ER-based MRI/MRSI information to guide prostate radiation therapy treatment. PMID- 15890606 TI - Initial clinical results of an in vivo dosimeter during external beam radiation therapy. AB - PURPOSE: An implantable radiation dosimeter has been developed to monitor dose delivered at depth in patients undergoing external beam therapy. A clinical pilot study was conducted to test the safety, efficacy, and utility of the device. METHODS AND MATERIALS: Ten patients, all with unresectable malignant disease, were enrolled to assess implantation risk and movement of the device in the body and to compare the in vivo measured dose to the value predicted by the treatment planning system software. RESULTS: Migration of the sensor away from the point of original placement was noted in only 1 patient (due to unconsolidated host tissue) and no adverse events were recorded during the implantation procedure or thereafter. Daily dose measurements were recorded successfully for all sensors in all patients. Variance between measured and predicted dose values was reported as a frequency of error at the > or =5% and > or =8% levels. The error frequency at the > or =8% level was as high as 47%, 29%, and 21% for lung, prostate, and rectal tumors, respectively. CONCLUSIONS: The implantable dosimeter was found to be safe and effective in measuring dose at depth. There are many factors that can influence delivered dose, and the implantable dosimeter measures the net effect of these factors. The daily sensor readings provide a new tool for rigorous treatment quality assurance. PMID- 15890607 TI - In regard to Dr. Souhami et al. (Int J Radiat Oncol Biol Phys 2004;60:853-860). PMID- 15890608 TI - Staging of advanced cervical cancer. PMID- 15890610 TI - Chasing your dural tail: Factors predicting local tumor control after gamma knife stereotactic radiosurgery for benign intracranial meningiomas: In regard to DiBiase et al. (Int J Radiat Oncol Biol Phys 2004;60:1515-1519). PMID- 15890612 TI - In regard to Dr. Ortholan et al. (Int J Radiat Oncol Biol Phys 2005;61:154-162). PMID- 15890615 TI - The plant multigenic family of thiol peroxidases. AB - Thiol peroxidases are ubiquitous recently characterized heme-free peroxidases, which catalyze the reduction of peroxynitrites and of various peroxides by catalytic cysteine residues and thiol-containing proteins as reductants. In plants, five different classes can be distinguished, according to the number and the position of conserved catalytic cysteines. Four classes are defined as peroxiredoxins and were already identified by phylogenetic sequence analysis, 1 Cys, 2-Cys, type II, and type Q peroxiredoxins, and the fifth is represented by glutathione peroxidases, which were recently shown to possess a thioredoxin dependent activity in plants. Since the discovery of peroxiredoxins in plants in 1996, a lot of work has been devoted to the biochemical and functional characterization of the different peroxiredoxin isoforms, but in contrast, few structural data are available. The analysis of the Arabidopsis thaliana genome indicates that at least 17 isoforms of thioredoxin-dependent peroxidases are expressed in various plant compartments. The role of these proteins is discussed in terms of electron donor and substrate specificities and in light of their expression and localization. These enzymes are expressed in many plant tissues and are involved notably in the protection of the photosynthetic apparatus, in the response to various biotic or abiotic stresses by fighting reactive oxygen or nitrogen species and lipid peroxidation. PMID- 15890616 TI - Peroxiredoxin 6, a 1-Cys peroxiredoxin, functions in antioxidant defense and lung phospholipid metabolism. AB - Peroxiredoxin 6 (Prdx6), a bifunctional 25-kDa protein with both GSH peroxidase and phospholipase A2 activities, is the only mammalian 1-Cys member of the peroxiredoxin superfamily and is expressed in all major organs, with a particularly high level in lung. Prdx6 uses GSH as an electron donor to reduce H2O2 and other hydroperoxides including phospholipid hydroperoxides at approximately 5 micromol/mg protein/min with K1 approximately 3 x 10(6) M(-1) s( 1). Oxidation of the Cys47 to a sulfenic acid during catalysis requires piGST catalyzed glutathionylation and reduction with GSH to complete the enzymatic cycle. Prdx6 stably overexpressed in cells protected against oxidative stress, whereas antisense treatment resulted in oxidant stress and apoptosis. Adenoviral mediated overexpression of Prdx6 in mouse lungs protected against the toxicity of hyperoxia, whereas Prdx6-null mice were more sensitive to the effects of hyperoxia or paraquat. We postulate that Prdx6 functions in antioxidant defense mainly by facilitating repair of damaged cell membranes via reduction of peroxidized phospholipids. The PLA2 activity of Prdx6 is Ca2+ independent and maximal at acidic pH. Inhibition of PLA2 activity results in alterations of lung surfactant phospholipid synthesis and turnover. Thus, Prdx6, a unique mammalian peroxiredoxin, is an important antioxidant enzyme and has a major role in lung phospholipid metabolism. PMID- 15890617 TI - Reactive oxygen species and the modulation of stroke. AB - Reactive oxygen species and oxidative state are slowly gaining acceptance in having a physiological relevance rather than just being the culprits in pathophysiological processes. The control of the redox environment of the cell provides for additional regulation in relation to critical cellular signal transduction pathways. Conversely, aberrant regulation of oxidative state manifesting as oxidative stress can predispose a cell to adverse outcome. The PI3 kinase/Akt pathway is one such pathway that is partially regulated via oxidative state and, in an oxidative stress paradigm such as ischemic reperfusion injury, may be inactivated, which can lead to potentiation and or exacerbation of cell death. Activation of NF(kappa)B has also been associated with oxidative stress. The role of NF(kappa)B in neuronal cell death is widely debated, with major studies highlighting both a pro- and an antiapoptotic role for NF(kappa)B with the outcome being region, stimulus, dose, and duration specific. This review hopes to make clear that oxidative state plays a key role in the regulation and control of numerous signal transduction pathways in the cell and that elucidating the mechanisms behind oxidative stress-mediated neuronal cell death is important in identifying potential putative targets for the treatment of neuropathologies such as stroke. PMID- 15890618 TI - Commentary on "Downregulation of the human Lon protease impairs mitochondrial structure and function and causes cell death" by D.A. Bota, J.K. Ngo, and K.J.A. Davies. PMID- 15890619 TI - Synergism of Helicobacter pylori infection and stress on the augmentation of gastric mucosal damage and its prevention with alpha-tocopherol. AB - Despite evidence that Helicobacter pylori (H. pylori) infection is closely associated with stress in gastric ulcer patients, the underlying mechanism why ulcer recurrence after stress is augmented especially in patients with H. pylori remains unknown. In this study, we found that oxidative stress played a critical role in the augmented mucosal damage provoked by water immersion restraint stress (WIRS) in H. pylori infection and that an antioxidant, alpha-tocopherol, could ameliorate the aggravation of stress-associated gastric mucosal damage. Two hundred forty SD rats were divided into two groups according to H. pylori inoculation, and after 24 weeks of H. pylori infection, the water immersion restraint stress was imposed for 30, 120, or 480 min, respectively. To evaluate the therapeutic effects of an antioxidant, alpha-tocopherol was administrated 40 mg/kg daily prior to imposing WIRS. Remarkably increased hemorrhagic lesions and bleeding indexes were noted in the H. pylori-infected group with statistical significance (P < 0.05) compared to the noninfected group at the same duration of WIRS. Significantly higher oxidative stress documented by iNOS, lipid peroxides, and GSH level was detected in gastric homogenates of the H. pylori-infected group. Proteomic analysis using 2-dimensional electrophoresis showed a decrease of HSP27 and other chaperone proteins. alpha-Tocopherol pretreatment significantly prevented the gastric mucosal damage, caused by WIRS in the presence of H. pylori. alpha-Tocopherol induced HSP27 expression, which was well correlated with downregulation of iNOS mRNA. Conclusively, the presence of H. pylori caused significant deterioration of stress-induced gastric mucosal lesions through increased oxidative stress and thus antioxidant treatment such as alpha tocopherol protected the gastric injuries. PMID- 15890620 TI - Heterozygous deficiency of manganese superoxide dismutase results in severe lipid peroxidation and spontaneous apoptosis in murine myocardium in vivo. AB - To circumvent the early lethality of manganese superoxide dismutase (SOD2) deficient mice, we have used a skin-specific strategy with introduction of loxP sites flanking exon 3 of the SOD2 gene. To our surprise, when breeding a female keratin 14 Cre transgenic mouse to a SOD2 "floxed" male mouse, due to keratin 14 promoter-driven Cre expression in the oocytes, all offspring were heterozygous for SOD2. In sharp contrast to initial publications on SOD2(+/-) mice, the herein reported mice on a mixed genetic background (C57BL/6 x 129/Ola) in their heterozygous state (SOD(+/-)) revealed distinct ultrastructural damage of the myocard, with swelling and disruption of mitochondria and accumulation of lipid droplets, increased nitrotyrosine formation, and lipid peroxidation as well as activation of apoptosis signaling pathways in the heart in vivo. Strikingly, and so far unreported, we found a substantial decrease in the activity of the cytosolic copper, zinc superoxide dismutase (SOD1) in the heart tissue of SOD2(+/ ) mice, suggesting that the breakdown of mitochondrial membranes in the heart of SOD2(+/-) mice results in the enhanced release of superoxide anion radicals or derivatives thereof with subsequent inactivation of cytosolic SOD1. This model may be particularly suited to long-term studies on age-related heart failure as well as other age-related diseases and the polygenic base of tissue-specific responses to oxidative injury. PMID- 15890621 TI - Valproic acid glucuronidation is associated with increases in 15-F2t-isoprostane in rats. AB - Oxidative stress has been associated with valproic acid (VPA) treatment in rats and studies are ongoing to examine the relationship between VPA biotransformation and the increase in the lipid peroxidation biomarker 15-F2t-isoprostane (15-F2t IsoP). This study investigated the effects of modulating VPA-1-O-acyl glucuronide (VPA-G) formation on 15-F2t-IsoP levels. Adult male Sprague-Dawley rats were pretreated with phenobarbital (PB; 80 mg/kg/day for 4 days), (-)-borneol (320 mg/kg), or a combination of both before VPA treatment (500 mg/kg). Liver VPA-G levels were determined by LC/MS and plasma and liver 15-F2t-IsoP levels were measured using an EIA method. PB, an inducer of VPA glucuronidation, elevated both liver VPA-G and plasma and liver 15-F2t-IsoP levels in VPA-treated rats. (-) Borneol, an inhibitor of glucuronidation, significantly reduced the levels of liver VPA-G and decreased plasma and liver 15-F2t-IsoP levels in both the VPA and the PB + VPA groups. (-)-Borneol and PB alone did not elevate 15-F2t-IsoP levels compared to the vehicle control groups. The fluorinated analogue of VPA, alpha fluoro-VPA, was a poor substrate for glucuronidation and did not elevate 15-F2t IsoP levels. In summary, the VPA-induced formation of 15-F2t-IsoP is apparently associated with VPA glucuronidation. PMID- 15890622 TI - Pyruvate but not lactate prevents NADH-induced myoglobin oxidation. AB - In this work, we investigated the influence of NADH on the redox state of myoglobin and the roles of pyruvate and lactate in this process. NADH increased the autoxidation rate of myoglobin. Both a drop in pH and partial deoxygenation markedly stimulated the autoxidation process and the influence of NADH. A correlation between met-Mb formation rate and NADH oxidation rate was always observed. The increased rate of Mb autoxidation caused by NADH was inhibited by catalase and pyruvate but not by l-lactate. The antioxidant activity versus H2O2 of both pyruvate and lactate was evidenced by chemiluminescence experiments. The antioxidant activity of lactate disappeared completely in the presence of myoglobin or apo-myoglobin, whereas it was only reduced for pyruvate. These results could be of interest in preventing autoxidation of myoglobin that can contribute to ischemia-reperfusion injury during infarction or high-intensity exercise. PMID- 15890623 TI - Identification of the main oxidation products of 8-methoxy-2'-deoxyguanosine by singlet molecular oxygen. AB - It is now well established that oxidation of 2'-deoxyguanosine (dGuo) in DNA by singlet molecular oxygen [O2 (1Delta(g))] produces 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo), whereas the main degradation products of free dGuo in aqueous solution have been identified as the two diastereomers of spiroiminodihydantoin nucleoside. Interestingly, O2 (1Delta(g))-mediated oxidation of free 8-oxodGuo gives rise to a pattern of degradation products that is different from that observed when the nucleoside is inserted into DNA. The reasons for these differences and the mechanisms involved in the oxidation reactions are not yet completely understood for either dGuo or 8-oxodGuo, either free or within DNA. In the present work, we report a study of the reaction of O2 (1Delta(g)) toward a modified nucleoside, 8-methoxy-2'-deoxyguanosine (8 MeOdGuo), either free or incorporated into an oligonucleotide. The reason for the choice of 8-MeOdGuo as a chemical model to study in more detail the oxidation pathways of 8-oxodGuo or, more precisely, of the tautomeric 8-hydroxy-2' deoxyguanosine was dictated by the fact that only the 7,8-enolic tautomer is present in the molecule. The thermolysis of an endoperoxide of a naphthalene derivative as a clean chemical source of 18O-labeled O2 (1Delta(g)) was used to oxidize 8-MeOdGuo. The main O2 (1Delta(g)) oxidation products that were separated and analyzed by HPLC coupled to tandem mass spectrometry were identified as the 2'-deoxyribonucleoside derivatives of 2,2,4-triamino-5-(2H)oxazolone, 2,5-diamino 4H-imidazol-4-one together with the methyl-substituted derivatives of spiroiminodihydantoin, oxidized iminoallantoin and urea. On the other hand, O2 (1Delta(g)) oxidation of 8-MeOdGuo-containing oligonucleotide generated imidazolone as the predominant degradation product. These results provided new mechanistic insights into the reactions of O2 (1Delta(g)) with purine nucleosides. PMID- 15890624 TI - S-glutathionylation in human platelets by a thiol-disulfide exchange-independent mechanism. AB - Protein-glutathione mixed disulfide formation was investigated in vitro by exposure of human platelets to the thiol-specific oxidant azodicarboxylic acid bis-dimethylamide (diamide). We found that diamide causes a decrease in the reduced form of glutathione (GSH), paralleled by an increase in protein-GSH mixed disulfides (S-glutathionylated proteins), which was not accompanied by any significant increase in the basal level of glutathione disulfide (GSSG). The increase in the appearance of S-glutathionylated proteins was inversely correlated with ADP-induced platelet aggregation. Platelet cytoskeleton was analyzed by SDS-PAGE followed by Western immunoblotting with anti-GSH antibody. The main S-glutathionylated cytoskeletal protein proved to be actin, which accounts for 35% of the platelet total protein content. Our results suggest that neither GSSG formation nor a consequent thiol-disulfide exchange mechanism is involved in actin S-glutathionylation of human platelets exposed to diamide. Instead, a mechanism involving the initial oxidative activation of actin thiol groups, which then react with GSH to the protein-GSH mixed disulfides, makes it likely that platelet actin is S-glutathionylated without any significant increase in the GSSG content. PMID- 15890625 TI - Nitration of the tyrosyl radical in ribonucleotide reductase by nitrogen dioxide: a gamma radiolysis study. AB - Nitrogen dioxide is a product of peroxynitrite homolysis and peroxidase-catalyzed oxidation of nitrite. It is of great importance in protein tyrosine nitration because most nitration pathways end with the addition of *NO2 to a one-electron oxidized tyrosine. The rate constant of this radical addition reaction is high with free tyrosine-derived radicals. However, little is known of tyrosine radicals in proteins. In this paper, we have used *NO2 generated by gamma radiolysis to study the nitration of the R2 subunit of ribonucleotide reductase, which contains a long-lived tyrosyl radical on Tyr122. Most of the nitration occurred on Tyr122, but nonradical tyrosines were also modified. In addition, peptidic bonds close to nitrated Tyr122 could be broken. Nitration at Tyr122 was not observed with a radical-free metR2 protein. The estimated rate constant of the Tyr122 radical reaction with *NO2 was of 3 x 10(4) M(-1) s(-1), thus several orders of magnitude lower than that of a radical on free tyrosine. Nitration rate of other tyrosine residues in R2 was even lower, with an estimated value of 900 M(-1) s(-1). This study shows that protein environment can significantly reduce the reactivity of a tyrosyl radical. In ribonucleotide reductase, the catalytically active radical residue is very efficiently protected against nitrogen oxide attack and subsequent nitration. PMID- 15890626 TI - A Caenorhabditis elegans mutant lacking functional nicotinamide nucleotide transhydrogenase displays increased sensitivity to oxidative stress. AB - Proton-translocating mitochondrial nicotinamide nucleotide transhydrogenase (NNT) was investigated regarding its physiological role in Caenorhabditis elegans. NNT catalyzes the reduction of NADP(+) by NADH driven by the electrochemical proton gradient, Deltap, and is thus a potentially important source of mitochondrial NADPH. Mitochondrial detoxification of reactive oxygen species (ROS) by glutathione-dependent peroxidases depends on NADPH for regeneration of reduced glutathione. Transhydrogenase may therefore be directly involved in the defense against oxidative stress. nnt-1 deletion mutants of C. elegans, nnt-1(sv34), were isolated and shown to grow essentially as wild type under normal laboratory conditions, but with a strongly lowered GSH/GSSG ratio. Under conditions of oxidative stress, caused by the superoxide-generating agent methyl viologen, growth of worms lacking nnt-1 activity was severely impaired. A similar result was obtained by using RNAi. Reintroducing nnt-1 in the nnt-1(sv34) knockout mutant led to a partial rescue of growth under oxidative stress conditions. These results provide evidence for the first time that nnt-1 is important in the defense against mitochondrial oxidative stress. PMID- 15890627 TI - Glutathione conjugates of 4-hydroxy-2(E)-nonenal as biomarkers of hepatic oxidative stress-induced lipid peroxidation in rats. AB - Here we present a simple, specific, and sensitive liquid chromatography/mass spectrometry method to measure 4-hydroxy-2(E)-nonenal-glutathione (HNE-GSH), the major stable hepatic metabolite of HNE after GSH conjugation, as a marker of oxidative stress in rat liver and hepatocytes. Commonly employed methods for the measurement of lipid peroxidation-derived free aldehydes or modified proteins suffer from the artificial formation of HNE or HNE adducts to cellular molecules during sample preparation and derivatization, resulting in an overestimation of background levels. Basal levels of HNE-GSH in liver tissue from untreated rats were detected in amounts of 20 pmol/g liver. Rats exposed to a single dose of iron nitrilotriacetate (Fe(III)NTA; 15 mg Fe/kg bw, ip), a model compound for the induction of oxidative stress, revealed a fivefold increase in the hepatic HNE GSH levels compared to controls 5 h after dosing. Moreover, a significant increase in HNE-mercapturic acid (HNE-MA) and its reduced metabolite DHN-MA was evident at 5 or 24 h after treatment, which was also reflected in increased plasma concentrations of these secondary HNE-GSH metabolites. In agreement with the in vivo data, a time-dependent increase in the levels of HNE-GSH from <1 to 123 +/- 16 pmol/10(6) cells over 5 h was detected in rat hepatocytes treated with Fe(III)NTA (150 microM). An increase in cellular HNE-GSH from <1.0 to 7.2 +/- 0.3 pmol/10(6) cells could be observed in rat hepatocytes treated with allyl alcohol (500 microM, 3 h), known for generation of HNE in hepatocytes. These data suggest that the direct measurement of the stable GSH conjugation product of cellular HNE in rat primary hepatocytes or its secondary metabolites may represent a reliable biomarker of oxidative stress-induced lipid peroxidation in rat liver in vivo. PMID- 15890628 TI - Isoprostanes in amniotic fluid: a predictive marker for fetal growth restriction in pregnancy. AB - Isoprostanes are markers of free radical-catalyzed lipid peroxidation. Evidence suggests that oxidative stress occurs in pregnancies with fetal growth restriction (FGR). The aim of this study was to analyze F2-isoprostanes in amniotic fluid of FGR pregnancies. We tested the hypothesis that F2-isoprostanes are reliable markers to distinguish FGR pregnancies from normal ones and appropriate-for-gestational-age (AGA) from small-for-gestational-age (SGA) newborns. F2-isoprostanes levels were measured by colorimetric enzyme immunoassay in the amniotic fluid of 77 pregnancies with normal fetal growth (group I) and 37 with FGR (group II). Fetal biometry and Doppler measurements were obtained using an ATL HDI 3000 ultrasound system. Isoprostanes were higher in group II than group I. The ROC curve distinguished group I from group II, showing 100% sensitivity and 88.3% specificity at a cutoff of 94 pg/ml. There were no statistical differences in isoprostanes levels between AGA and SGA newborns in group II. The area under the ROC curve drawn to distinguish AGA and SGA newborns showed a sensitivity of 100% and a specificity of 72.3% at a cutoff of 94 pg/ml. The relative risk index indicated a 8.05 times higher risk of birth weight below the 3rd percentiles in group II than in group I. High isoprostanes concentrations can be detected in the amniotic fluid of FGR pregnancies and the assay of isoprostanes in amniotic fluid is a reliable assessment of fetal oxidative stress. Common use of this predictive marker in obstetrics will improve the ability of clinicians to identify those fetuses who will be born SGA or with a birth weight below the 25th percentile. PMID- 15890629 TI - What can be learned by assessing funding patterns for pain research? A commentary on National Institutes of Health grant awards for pain, nausea, and dyspnea research. PMID- 15890630 TI - National Institutes of Health grant awards for pain, nausea, and dyspnea research: an assessment of funding patterns in 2003. AB - We introduce an interactive database that permits description and exploration of National Institutes of Health (NIH) funding patterns for research on pain, nausea, and dyspnea. The database encompasses both basic science and clinical research. This article describes how we created the database, including the procedures we developed for reviewing and classifying research grants. In addition, it reports NIH grants and funding activity for the year 2003, with a breakdown of funding activity by Institute and funding comparisons across Institutes. It also describes a first attempt to identify clinically significant but underfunded research domains. In 2003, the NIH funded 1148 grants having relevance to the domain of pain, representing 2.5% of the total NIH research budget. Of those, 581 grants, or about 1% of the NIH budget, had a primary focus on pain. Of the diseases and conditions addressed by the current implementation, musculoskeletal conditions were the best represented with 105 grants, whereas cardiac conditions had the fewest number of grants with 7. The NIH funded 43 grants for dyspnea research and a scant 29 grants for nausea studies. We discuss the current limitations of the database and our plans for further development. PERSPECTIVE: The interactive database and classification system for pain, nausea, and dyspnea research funded by the NIH reported on in this article represents an objective and verifiable resource for health policy makers and others interested in NIH funding decisions. The high inter-rater reliability achieved attests to the objectivity of the classification method. Initial analyses demonstrate that these data can usefully track funding patterns by NIH institutes and reveal underfunded areas of research. PMID- 15890631 TI - The role of fear of movement and injury in selective attentional processing in patients with chronic low back pain: a dot-probe evaluation. AB - The present study sought to investigate to what extent patients with chronic low back pain and pain-free control subjects selectively attend to pain-related stimuli as measured with 2 dot-probe tasks with word stimuli and pictorial stimuli. Selective attentional processing was measured by means of 3 indices: the bias index, a congruency effect, and an incongruency effect. Pain-related fear as a trait measure (Tampa Scale for Kinesiophobia [TSK]) was expected to be positively associated with all indices of selective attentional processing of pain stimuli. Results were analyzed with repeated-measures analysis of variance. An incongruency effect was found for patients and to a significantly less degree for pain-free control subjects on the dot-probe task with pictorial stimuli, indicating that pain patients have difficulty disengaging from threat pictures. Pain-related fear as a trait measure (TSK) was not associated with selective attentional processing of word and pictorial stimuli in either pain patients or control subjects. Results from the present study are discussed, and directions for future research are provided. PERSPECTIVE: Demonstrating difficulty to disengage from threat might be clinically relevant because patients might pay less attention to fear-disconfirming information and remain engaged in avoidance, which might eventually lead to prolonged anxiety states. PMID- 15890632 TI - Access to care for chronic pain: racial and ethnic differences. AB - Access to medical care is a major national issue, and several surveys suggest that racial and ethnic differences influence access to care for chronic pain problems. To evaluate the influence of race and ethnicity on access to treatment for chronic pain, a cross-sectional telephone survey was performed in a nationally representative sample of 454 white, 447 African-American, and 434 Hispanic subjects with pain for > or =3 months. Questions explored demographics, pain and its treatment, and perceived access to care. A composite "access" variable combined actual consultation with perceived access. Hispanics were younger, least likely to be insured, and had the least education and lowest income; 61% spoke Spanish at home. Hispanics were significantly less likely to have consulted a primary care practitioner for pain (70%) than whites (84%) or African-Americans (85%). A lower likelihood of consultation also was associated with speaking Spanish, being male, being relatively young (18-34 years old) or single, having limited education, and not being employed. Low "access" to care was associated with being Hispanic and speaking Spanish, being younger or male, having low income or limited education, being employed, and agreeing that financial concerns prevented pain treatment. High "access" was associated with being white or African-American; being older or female or living in a suburban area; having insurance, higher income, or college education; and being unemployed. In multivariate models, low "access" was associated with Hispanic ethnicity and agreement that financial concerns prevented pain treatment. High "access" was associated with more severe pain, having insurance or an income of US 25,000 dollars to US 74,000 dollars, and agreeing that "A doctor or other health care provider is the first person I would go to to discuss my pain." These data suggest that race/ethnicity, other demographic characteristics, and socioeconomic factors influence access to pain care. Hispanic ethnicity predicts limited access. PERSPECTIVE: The influence of race and ethnicity on access to health care is a major issue in the United States. A national telephone survey suggests that race and ethnicity, along with other demographic and socioeconomic factors, influence access to care for chronic pain. PMID- 15890633 TI - Three distinct categories of time course of pain produced by oral capsaicin. AB - Humans vary in oral pain tolerance. Our earlier studies noted that the responses of subjects show 1 of 3 qualitative response patterns to a single oral capsaicin concentration, which we termed a tonic pattern (level detection response), a phasic pattern (change detection response), and an integrator pattern (cumulative irritation) response. These patterns were modeled quantitatively as the sum of 3 underlying processes. Two time-varying capsaicin stimulus profiles were designed from the quantitative model. In the ascending step paradigm, 30 ppm capsaicin was presented to 42 subjects for 15 minutes, followed immediately and without explanation by 300 ppm capsaicin for 25 minutes. In the descending step paradigm, 300 ppm capsaicin was presented to 36 other subjects for 24 minutes, followed by 10 ppm for 22 minutes. Subjective burn was rated at 1 minute and then at 3-minute intervals throughout the presentation. Fuzzy cluster analysis identified 3 distinct response phenotypes in each paradigm, corresponding to level detection, change detection, and cumulative irritation response patterns identified previously. Discriminant functions permitted classification of these phenotypes from the response patterns. Thus, these paradigms provide the first quantitative phenotypic description of distinct oral pain responses to a common irritant, capsaicin. PERSPECTIVE: This study examined the time-dependent behavior of pain produced by oral application of capsaicin. Three distinct temporal response phenotypes were identified objectively: level detection, change detection, and cumulative irritation detection. These time-dependent analyses provide a new dimension to understanding individual differences in pain sensation in clinical settings. PMID- 15890634 TI - Effects of the N-methyl-D-aspartate receptor antagonist dextromethorphan on temporal summation of pain are similar in fibromyalgia patients and normal control subjects. AB - Temporal summation of second pain at least partly reflects temporal summation of dorsal horn neuronal responses, and both have been termed windup (WU), a form of nociception-dependent central sensitization. Animal and human experiments have shown that both forms of WU depend on N-methyl-D-aspartate (NMDA) and substance P receptor systems. WU of second pain (WU(SP)) in patients with fibromyalgia (FM) is enhanced compared with normal control (NC) subjects and is followed by exaggerated WU(SP) aftersensations and prolonged WU(SP) maintenance at low stimulus frequencies. Because the enhanced WU(SP) of FM patients could be related to abnormal endogenous modulation of NDMA receptors, we tested the effects of the NMDA receptor antagonist dextromethorphan (DEX) on WU(SP) in FM and NC subjects in a double-blind, placebo-controlled, crossover study. WU(SP) was elicited by trains of 0.7-second duration thermal pulses applied to the glabrous surface of the hands or by 1-second mechanical stimuli to the adductor pollicis muscle of the hands at a frequency of 0.33 Hz. In comparison to baseline and placebo conditions, single oral doses of DEX 60 and 90 mg reduced thermal and mechanical WU(SP) in NC and FM subjects, with DEX 90 mg being most effective. These effects did not differ for male and female NC subjects. FM subjects required less thermal and mechanical stimulus intensity than NC to achieve maximal WU(SP), but the extent of WU(SP) reduction by DEX did not statistically differ between NC and FM subjects for all study conditions. Thus, central pain processing of FM subjects is not different from NC in at least one important aspect, namely their NMDA receptor system responsiveness to pharmacologic inhibition by DEX. PERSPECTIVE: Results of this study demonstrate that FM patients show abnormal WU(SP) during thermal and mechanical stimulation compared with NC. Because oral doses of the NMDA receptor antagonist DEX attenuated thermal and mechanical WU(SP) similarly in FM patients and NC, other mechanisms than WU(SP) need to be considered for the widespread pain of FM patients. These mechanisms might include tonic nociceptive input from peripheral tissues and/or enhanced descending facilitation. PMID- 15890635 TI - Botulinum toxin a injection of the obturator internus muscle for chronic perineal pain. AB - Chronic perineal pain is often a difficult condition to manage. Current treatments include pudendal nerve injections and pudendal nerve release surgery. The obturator internus muscle has a close relationship to the pudendal nerve and might be a potential target for therapeutic intervention. PERSPECTIVE: A case is presented of refractory perineal pain that was successfully treated by injecting the obturator internus muscle with botulinum toxin A. PMID- 15890636 TI - Catastrophizing and experimental pain sensitivity: only in vivo reports of catastrophic cognitions correlate with pain responses. PMID- 15890637 TI - Operative stabilization of posterior shoulder instability. AB - BACKGROUND: Symptomatic, traumatic posterior shoulder instability is often the result of a posteriorly directed blow to an adducted, internally rotated, and forward-flexed upper extremity. Operative repair has been shown to provide favorable results. Current arthroscopic techniques with suture anchors and the ability to plicate the capsule using a nonabsorbable suture may provide favorable outcomes with reduced morbidity. PURPOSE: To evaluate the results of operative shoulder stabilization in patients with traumatic posterior shoulder instability. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: A consecutive series of patients who underwent arthroscopic or open posterior stabilization for traumatic posterior shoulder instability were evaluated using subjective assessments, physical examinations, the Single Assessment Numeric Evaluation, Rowe score, Simple Shoulder Test, and the Western Ontario Shoulder Instability Index. RESULTS: Between May 1996 and February 2002, 31 shoulders (30 patients) underwent posterior stabilization (19 arthroscopically, 12 open). There were 29 men and 1 woman (mean age, 23 years). Preoperatively, all patients had a distinct traumatic cause for the instability. On physical examination, all patients had posterior apprehension and increased (2+, 3+) posterior load-shift testing. Preoperative radiographs and/or magnetic resonance imaging revealed posterior rim calcification or reverse Bankart lesions in 29 cases (94%). At arthroscopy, posterior labral injuries, reverse Bankart lesions, or humeral head defects were identified. Follow-up averaged 40 months, and the mean duration between injury and surgery was 21 months. The mean Single Assessment Numeric Evaluation, Rowe score, Simple Shoulder Test, and Western Ontario Shoulder Instability Index scores, respectively, for the entire group were 89, 87, 11, and 346; for the open group, they were 81, 80, 10.5, and 594; for the arthroscopic group, they were 92, 92, 11.4, and 190. The Western Ontario Shoulder Instability Index (P < .03) and Rowe score (P < .04) outcomes scores for the arthroscopic group were statistically better than those of the open group. Twenty-nine of 31 shoulders were rated as excellent or good. CONCLUSION: In the case of traumatic posterior shoulder subluxation, posterior lesions of the labrum ("reverse Bankart"), articular edge, and capsule are observed. Surgical treatment addressing these lesions led to satisfactory results for both the open and arthroscopic treated groups. In this study, an arthroscopic technique utilizing suture anchor repair with capsular placation provided the most favorable outcomes. PMID- 15890638 TI - Aromatase inhibitors as adjuvant therapy for postmenopausal patients with early stage breast cancer. AB - Endocrine therapy of hormone receptor-positive breast tumors is widely used as palliative therapy for metastatic breast cancer and as adjuvant therapy for early stage breast cancer. Tamoxifen has been the definitive standard of hormonal therapies for the last 30 years because of its documented efficacy and reasonable safety profile. Based on encouraging results from trials utilizing the selective, third generation aromatase inhibitors (AIs) in metastatic breast cancer, a number of trials were designed to examine these agents as adjuvant therapies. Trials directly comparing AIs with tamoxifen have, to date, demonstrated superior disease-free-survival with AIs. Likewise, trials examining the use of AIs after tamoxifen have demonstrated better outcomes compared with tamoxifen alone. Additionally, letrozole has been demonstrated to result in superior disease-free survival after 5 years of adjuvant tamoxifen, compared with no further therapy. In general, the AIs are tolerated at least as well as tamoxifen but decrease bone mineral density and increase osteoporosis due to their lack of estrogenic effects on bone. Based on the fact that AIs appear more effective at preventing contralateral breast cancers than tamoxifen, they are being examined as breast cancer preventives. Despite available data using the AIs as adjuvant therapies, many questions remain unanswered, and further trials will be needed to address these important issues. PMID- 15890639 TI - Approaching difficult communication tasks in oncology. AB - Effective communication skills enable physicians to improve patients' understanding of their illnesses, improve patient adherence to treatment regimens, use time efficiently, avoid burnout, and increase professional fulfillment. Common communication pitfalls include blocking, lecturing, depending on a routine, collusion, and premature reassurance. Fundamental communication skills include "ask-tell-ask," "tell me more," and responding empathetically. Key communication tasks that are linked to the illness trajectory include: the first visit, giving bad news, making anticancer treatment decisions, offering clinical trials, completing anticancer therapy, and discontinuing palliative chemotherapy. While enhancing or acquiring new skills ultimately requires practice and feedback over time, this article provides a cognitive map for important communication skills that physicians need over the course of caring for a person with cancer. PMID- 15890640 TI - Targeting apoptosis pathways in cancer therapy. AB - Apoptosis, or programmed cell death, is a mechanism by which cells undergo death to control cell proliferation or in response to DNA damage. The understanding of apoptosis has provided the basis for novel targeted therapies that can induce death in cancer cells or sensitize them to established cytotoxic agents and radiation therapy. These novel agents include those targeting the extrinsic pathway such as tumor necrosis factor-related apoptosis-inducing ligand receptor 1, and those targeting the intrinsic Bcl-2 family pathway such as antisense bcl-2 oligonucleotides. Many pathways and proteins control the apoptosis machinery. Examples include p53, the nuclear factor kappa B, the phosphatidylinositol 3 kinase pathway, and the ubiquitin/proteosome pathway. These can be targeted by specific modulators such as bortezomib, and mammalian target of rapamycin inhibitors such as CCI-779 and RAD 001. Because these pathways may be preferentially altered in tumor cells, there is potential for a selective effect in tumors sparing normal tissue. This article reviews the current understanding of the apoptotic pathways, including the extrinsic (cytoplasmic) and intrinsic (mitochondrial) pathways, and the agents being developed to target these pathways. PMID- 15890641 TI - Hormonal therapy for breast cancer. PMID- 15890642 TI - Engineering conformational destabilization into mouse apolipoprotein E. A model for a unique property of human apolipoprotein E4. AB - Apolipoprotein (apo) E4 is a major risk factor for Alzheimer and cardiovascular diseases. ApoE4 differs from the two other common isoforms (apoE2 and apoE3) by its lower resistance to denaturation and greater propensity to form partially folded intermediates. As a first step to determine the importance of stability differences in vivo, we reengineered a partially humanized variant of the amino terminal domain of mouse apoE (T61R mouse apoE) to acquire a destabilized conformation like that of apoE4. For this process, we determined the crystal structure of wild-type mouse apoE, which, like apoE4, forms a four-helix bundle, and identified two structural differences in the turn between helices 2 and 3 and in the middle of helix 3 as potentially destabilizing sites. Introducing mutations G83T and N113G at these sites destabilized the mouse apoE conformation. The mutant mouse apoE more rapidly remodeled phospholipid than T61R mouse apoE, which supports the hypothesis that a destabilized conformation promotes apoE4 lipid binding. PMID- 15890643 TI - The pro-atherogenic cytokine interleukin-18 induces CXCL16 expression in rat aortic smooth muscle cells via MyD88, interleukin-1 receptor-associated kinase, tumor necrosis factor receptor-associated factor 6, c-Src, phosphatidylinositol 3 kinase, Akt, c-Jun N-terminal kinase, and activator protein-1 signaling. AB - We recently demonstrated that the chemokine CXCL16 is expressed in aortic smooth muscle cells (ASMC) and induces ASMC adhesion and proliferation (Chandrasekar, B., Bysani, S., and Mummidi, S. (2004) J. Biol. Chem. 279, 3188-3196). Here we reort that interleukin (IL)-18 positively regulates CXCL16 transcription in rat ASMC. We characterized the cis-regulatory region of CXCL16 and identified a functional activator protein-1 (AP-1) binding motif. Deletion or mutation of this site attenuated IL-18-mediated CXCL16 promoter activity. Gel shift, supershift, and chromatin immunoprecipitation assays confirmed AP-1-dependent CXCL16 expression. CXCL16 promoter-reporter activity was increased by constitutively active c-Fos and c-Jun and decreased by dominant negative or antisense c-Fos and c-Jun. Src kinase inhibitors PP1 and PP2, phosphatidylinositol 3-kinase (PI3K) inhibitors wortmannin and LY294002, Akt inhibitor, the c-Jun N-terminal kinase (JNK) inhibitor SP600125, antisense JNK and dominant negative MyD88, interleukin 1 receptor-associated kinase (IRAK)-1, IRAK4, and phosphatidylinositol 3-kinase expression all attenuated IL-18-mediated AP-1 binding and reporter activity, CXCL16 promoter-reporter activity, and CXCL16 expression. Thus IL-18 induced CXCL16 expression via a MyD88 --> IRAK1-IRAK4-TRAF6 (tumor necrosis factor receptor-associated factor 6) --> c-Src--> PI3K --> Akt --> JNK --> AP-1 pathway. Importantly, IL-18 stimulated ASMC proliferation in a CXCL16-dependent manner. These data provide for the first time a mechanism of IL-18-mediated CXCL16 gene transcription and CXCL16-dependent ASMC proliferation and suggest a role for IL 18-CXCL16 cross-talk in atherogenesis and restenosis following angioplasty. PMID- 15890644 TI - Physical and kinetic interactions between glutamyl-tRNA reductase and glutamate-1 semialdehyde aminotransferase of Chlamydomonas reinhardtii. AB - In plants, algae, and most bacteria, the heme and chlorophyll precursor 5 aminolevulinic acid (ALA) is formed from glutamate in a three-step process. First, glutamate is ligated to its cognate tRNA by glutamyl-tRNA synthetase. Activated glutamate is then converted to a glutamate 1-semialdehyde (GSA) by glutamyl-tRNA reductase (GTR) in an NADPH-dependent reaction. Subsequently, GSA is rearranged to ALA by glutamate-1-semialdehyde aminotransferase (GSAT). The intermediate GSA is highly unstable under physiological conditions. We have used purified recombinant GTR and GSAT from the unicellular alga Chlamydomonas reinhardtii to show that GTR and GSAT form a physical and functional complex that allows channeling of GSA between the enzymes. Co-immunoprecipitation and sucrose gradient ultracentrifugation results indicate that recombinant GTR and GSAT enzymes specifically interact. In vivo cross-linking results support the in vitro results and demonstrate that GTR and GSAT are components of a high molecular mass complex in C. reinhardtii cells. In a coupled enzyme assay containing GTR and wild-type GSAT, addition of inactive mutant GSAT inhibited ALA formation from glutamyl-tRNA. Mutant GSAT did not inhibit ALA formation from GSA by wild-type GSAT. These results suggest that there is competition between wild-type and mutant GSAT for binding to GTR and channeling GSA from GTR to GSAT. Further evidence supporting kinetic interaction of GTR and GSAT is the observation that both wild-type and mutant GSAT stimulate glutamyl-tRNA-dependent NADPH oxidation by GTR. PMID- 15890645 TI - The Pro335 --> Leu polymorphism of type 3 inositol 1,4,5-trisphosphate receptor found in mouse inbred lines results in functional change. AB - Inositol 1,4,5-trisphosphate receptor (IP3R) is an intracellular Ca2+ channel involved in various cellular signaling. Type 3 IP3R (IP3R3) retains ligand-gated Ca2+ channel properties differing from other subtypes in terms of IP3-binding affinity and regulation of its channel activity by effector molecules. In this study, we found the natural Pro335 --> Leu polymorphism of mouse IP3R3 between BALB/c and C57BL/6J. We investigated the functional differences between Pro335IP3R3 and Leu335IP3R3 with purified receptors reconstituted into proteoliposomes as well as with soluble ligand binding domains. Pro335IP3R3 exhibited significantly higher IP3-binding affinity and IP3-induced Ca2+ release than those of Leu335IP3R3 in both forms of the receptor. Moreover, the polymorphic change caused differences in the effect of external Ca2+ on IP3 induced Ca2+ release. The Pro335 --> Leu substitution alters the conformation of soluble ligand binding domain as revealed by intrinsic fluorescence and circular dichroism spectra with or without Ca2+. The results indicate that the polymorphism of IP3R3 causes changes in receptor function, presumably affecting intracellular Ca2+ signaling. PMID- 15890646 TI - Glycosphingolipid accumulation inhibits cholesterol efflux via the ABCA1/apolipoprotein A-I pathway: 1-phenyl-2-decanoylamino-3-morpholino-1 propanol is a novel cholesterol efflux accelerator. AB - Cellular glycosphingolipid (GSL) storage is known to promote cholesterol accumulation. Although physical interactions between GSLs and cholesterol are thought to cause intracellular cholesterol "trapping," it is not known whether cholesterol homeostatic mechanisms are also impaired under these conditions. ApoA I-mediated cholesterol efflux via ABCA1 (ATP-binding cassette transporter A1) is a key regulator of cellular cholesterol balance. Here, we show that apoA-I mediated cholesterol efflux was inhibited (by up to 53% over 8 h) when fibroblasts were treated with lactosylceramide or the glucocerebrosidase inhibitor conduritol B epoxide. Furthermore, apoA-I-mediated cholesterol efflux from fibroblasts derived from patients with genetic GSL storage diseases (Fabry disease, Sandhoff disease, and GM1 gangliosidosis) was impaired compared with control cells. Conversely, apoA-I-mediated cholesterol efflux from fibroblasts and cholesterol-loaded macrophage foam cells was dose-dependently stimulated (by up to 6-fold over 8 h) by the GSL synthesis inhibitor 1-phenyl-2-decanoylamino-3 morpholino-1-propanol (PDMP). Unexpectedly, a structurally unrelated GSL synthesis inhibitor, N-butyldeoxynojirimycin, was unable to stimulate apoA-I mediated cholesterol efflux despite achieving similar GSL depletion. PDMP was found to up-regulate ABCA1 mRNA and protein expression, thereby identifying a contributing mechanism for the observed acceleration of cholesterol efflux to apoA-I. This study reveals a novel defect in cellular cholesterol homeostasis induced by GSL storage and identifies PDMP as a new agent for enhancing cholesterol efflux via the ABCA1/apoA-I pathway. PMID- 15890647 TI - Phospholipids as determinants of membrane protein topology. Phosphatidylethanolamine is required for the proper topological organization of the gamma-aminobutyric acid permease (GabP) of Escherichia coli. AB - Evidence is accumulating that the topological organization and hence function of some membrane proteins are not solely determined by the amino acid sequence of the protein but are also influenced by the lipid composition of the membrane. The gamma-aminobutyric acid (GABA) permease (GabP) of Escherichia coli has been found in this study to be affected both topologically and kinetically by membrane lipids. Using single cysteine accessibility methods with viable E. coli strains of natural lipid composition and those lacking phosphatidylethanolamine (PE), we have shown that the N-terminal hairpin of GabP is inverted relative to the membrane in PE-lacking cells, with a hinge point in transmembrane domain III. The rate of GABA transport is reduced by more than 99% in PE-lacking cells. The Michaelis constant for GABA transport is not greatly affected nor is the dependence of transport on energy. However, "transport specificity ratio" analysis demonstrated a clear transition state stability difference for GABA and nipecotic acid between the protein in PE-containing and PE-lacking cells. The patterns of observed effects are similar to those seen with the phenylalanine transporter of E. coli (Zhang, W., Bogdanov, M. Pi, J. Pittard, A. J., and Dowhan, W. (2003) J. Biol. Chem. 278, 50128-50135), also an amino acid/polyamine/organocation family member but quite distinct from those observed with lactose permease (Bogdanov, M., Heacock, P. N., and Dowhan, W. (2002) EMBO J. 21, 2107-2116), a major facilitator superfamily member. Therefore, by extending the studies of similarities and differences in lipid responses among and between family groups, we may identify elements within the proteins that facilitate lipid responsiveness. PMID- 15890648 TI - Nuclear localization and mitogen-activated protein kinase phosphorylation of the multifunctional protein CAD. AB - CAD is a multifunctional protein that initiates and regulates mammalian de novo pyrimidine biosynthesis. The activation of the pathway required for cell proliferation is a consequence of the phosphorylation of CAD Thr-456 by mitogen activated protein (MAP) kinase. Although most of the CAD in the cell was cytosolic, cell fractionation and fluorescence microscopy showed that Thr(P)-456 CAD was primarily localized within the nucleus in association with insoluble nuclear substructures, including the nuclear matrix. CAD in resting cells was cytosolic and unphosphorylated. Upon epidermal growth factor stimulation, CAD moved to the nucleus, and Thr-456 was found to be phosphorylated. Mutation of the CAD Thr-456 and inhibitor studies showed that nuclear import is not mediated by MAP kinase phosphorylation. Two fluorescent CAD constructs, NLS-CAD and NES-CAD, were prepared that incorporated strong nuclear import and export signals, respectively. NLS-CAD was exclusively nuclear and extensively phosphorylated. In contrast, NES-CAD was confined to the cytoplasm, and Thr-456 remained unphosphorylated. Although alternative explanations can be envisioned, it is likely that phosphorylation occurs within the nucleus where much of the activated MAP kinase is localized. Trapping CAD in the nucleus had a minimal effect on pyrimidine metabolism. In contrast, when CAD was excluded from the nucleus, the rate of pyrimidine biosynthesis, the nucleotide pools, and the growth rate were reduced by 21, 36, and 60%, respectively. Thus, the nuclear import of CAD appears to promote optimal cell growth. UMP synthase, the bifunctional protein that catalyzes the last two steps in the pathway, was also found in both the cytoplasm and nucleus. PMID- 15890649 TI - Functional diversity of Csk, Chk, and Src SH2 domains due to a single residue variation. AB - The C-terminal Src kinase (Csk) family of protein tyrosine kinases contains two members: Csk and Csk homologous kinase (Chk). Both phosphorylate and inactivate Src family kinases. Recent reports suggest that the Src homology (SH) 2 domains of Csk and Chk may bind to different phosphoproteins, which provides a basis for different cellular functions for Csk and Chk. To verify and characterize such a functional divergence, we compared the binding properties of the Csk, Chk, and Src SH2 domains and investigated the structural basis for the functional divergence. First, the study demonstrated striking functional differences between the Csk and Chk SH2 domains and revealed functional similarities between the Chk and Src SH2 domains. Second, structural analysis and mutagenic studies revealed that the functional differences among the three SH2 domains were largely controlled by one residue, Glu127 in Csk, Ile167 in Chk, and Lys200 in Src. Mutating these residues in the Csk or Chk SH2 domain to the Src counterpart resulted in dramatic gain of function similar to Src SH2 domain, whereas mutating Lys200 in Src SH2 domain to Glu (the Csk counterpart) resulted in loss of Src SH2 function. Third, a single point mutation of E127K rendered Csk responsive to activation by a Src SH2 domain ligand. Finally, the optimal phosphopeptide sequence for the Chk SH2 domain was determined. These results provide a compelling explanation for the functional differences between two homologous protein tyrosine kinases and reveal a new structure-function relationship for the SH2 domains. PMID- 15890650 TI - Carbon Source-dependent assembly of the Snf1p kinase complex in Candida albicans. AB - The Snf1p/AMP-activated kinases are involved in transcriptional, metabolic, and developmental regulation in response to stress. In Saccharomyces cerevisiae, Snf1p (Cat1p) is one of the key regulators of carbohydrate metabolism, and cat1 (snf1) mutants fail to grow with non-fermentable carbon sources. In Candida albicans, Snf1p is an essential protein and cells depend on a functional Snf1 kinase even with glucose as carbon source. We investigated the CaSnf1p complex after tandem affinity purification and mass spectrometric analysis and show that the complex composition changes with the carbon source provided. Three subunits were identified, one of which was named CaSnf4p because of its homology to the ScSnf4 protein and the respective CaSNF4 gene could complement a S. cerevisiae snf4 mutant. The other two proteins revealed similarities to the S. cerevisiae kinase beta subunits ScGal83p, ScSip2p, and ScSip1p. Both genes complemented the scaffold function in a S. cerevisiae gal83,sip1,sip2 triple deletion mutant and were named according to their scaffold function as CaKIS1p and CaKIS2p. Matrix assisted laser desorption ionization peptide mass fingerprint analysis indicated that CaKis2p is N-terminal myristoylated and the incorporation of CaKis2p in the Snf1p complex was reduced when compared with cells grown with glucose as a carbon source. To verify the different complex assemblies, a stable isotope labeling technique (iTraqtrade mark) was employed, confirming a 3-fold decrease of CaKis2p with ethanol. Yeast two-hybrid analysis confirmed the interaction partners, and these results showed an activator domain for the CaKis2 protein that has not been reported for S. cerevisiae scaffold subunits. PMID- 15890651 TI - Energetic and structural consequences of perturbing Gly-193 in the oxyanion hole of serine proteases. AB - The oxyanion hole of serine proteases is formed by the backbone N atoms of the catalytic Ser-195 and Gly-193 and engages the backbone O atom of the P1 residue of substrate in an important H-bonding interaction. The energetic contribution of this interaction in the ground and transition states is presently unknown. Measurements of the individual rate constants defining the catalytic mechanism of substrate hydrolysis for wild-type thrombin and trypsin and their G193A and G193P mutants reveal that Gly-193 is required for optimal substrate binding and acylation. Crystal structures of the G193A and G193P mutants of thrombin bound to the active site inhibitor H-d-Phe-Pro-Arg-CH2Cl document the extent of perturbation induced by the replacement of Gly-193. The Ala mutant weakens the H bonding interaction of the N atom of residue 193, whereas the Pro substitution abrogates it altogether with additional small shifts of the protein backbone. From the kinetic and structural data, we estimate that the H-bonding interaction in the oxyanion hole contributes a stabilization of the ground and transition states of > 1.5 kcal/mol but < 3.0 kcal/mol. These results shed light on a basic aspect of the enzyme-substrate interaction in the entire family of trypsin-like serine proteases. PMID- 15890652 TI - The 1.3 A crystal structure of the flavoprotein YqjM reveals a novel class of Old Yellow Enzymes. AB - Here we report the crystal structure of YqjM, a homolog of Old Yellow Enzyme (OYE) that is involved in the oxidative stress response of Bacillus subtilis. In addition to the oxidized and reduced enzyme form, the structures of complexes with p-hydroxybenzaldehyde and p-nitrophenol, respectively, were solved. As for other OYE family members, YqjM folds into a (alpha/beta)8-barrel and has one molecule of flavin mononucleotide bound non-covalently at the COOH termini of the beta-sheet. Most of the interactions that control the electronic properties of the flavin mononucleotide cofactor are conserved within the OYE family. However, in contrast to all members of the OYE family characterized to date, YqjM exhibits several unique structural features. For example, the enzyme exists as a homotetramer that is assembled as a dimer of catalytically dependent dimers. Moreover, the protein displays a shared active site architecture where an arginine finger (Arg336) at the COOH terminus of one monomer extends into the active site of the adjacent monomer and is directly involved in substrate recognition. Another remarkable difference in the binding of the ligand in YqjM is represented by the contribution of the NH2-terminal Tyr28 instead of a COOH terminal tyrosine in OYE and its homologs. The structural information led to a specific data base search from which a new class of OYE oxidoreductases was identified that exhibits a strict conservation of active site residues, which are critical for this subfamily, most notably Cys26, Tyr28, Lys109, and Arg336. Therefore, YqjM is the first representative of a new bacterial subfamily of OYE homologs. PMID- 15890653 TI - SNAP-29-mediated modulation of synaptic transmission in cultured hippocampal neurons. AB - Identifying the molecules that regulate both the recycling of synaptic vesicles and the SNARE components required for fusion is critical for elucidating the molecular mechanisms underlying synaptic plasticity. SNAP-29 was initially isolated as a syntaxin-binding and ubiquitously expressed protein. Previous studies have suggested that SNAP-29 inhibits SNARE complex disassembly, thereby reducing synaptic transmission in cultured superior cervical ganglion neurons in an activity-dependent manner. However, the role of SNAP-29 in regulating synaptic vesicle recycling and short-term plasticity in the central nervous system remains unclear. In the present study, we examined the effect of SNAP-29 on synaptic transmission in cultured hippocampal neurons by dual patch clamp whole-cell recording, FM dye imaging, and immunocytochemistry. Our results demonstrated that exogenous expression of SNAP-29 in presynaptic neurons significantly decreased the efficiency of synaptic transmission after repetitive firing within a few minutes under low and moderate frequency stimulations (0.1 and 1 Hz). In contrast, SNAP-29 did not affect the density of synapses and basal synaptic transmission. Whereas neurotransmitter release was unaffected during intensive stimulation, recovery after synaptic depression was impaired by SNAP-29. Furthermore, knockdown of SNAP-29 expression in neurons by small interfering RNA increased the efficiency of synaptic transmission during repetitive firing. These findings suggest that SNAP-29 acts as a negative modulator for neurotransmitter release, probably by slowing recycling of the SNARE-based fusion machinery and synaptic vesicle turnover. PMID- 15890654 TI - A novel anti-platelet monoclonal antibody (3C7) specific for the complex of integrin alpha IIb beta3 inhibits platelet aggregation and adhesion. AB - Activation or ligand binding induces conformational changes in alpha IIb beta3, resulting in exposure of neoepitopes named ligand-induced binding sites. We reported here a novel monoclonal antibody developed by using Chinese hamster ovary (CHO) cells expressing an activated alpha IIb beta3 mutant (CHO alpha IIb beta3Delta717) as the immunogen. This IgG 2b kappa named 3C7 was specific for the complex of alpha IIb beta3 as demonstrated by flow cytometry, immunoprecipitation, and EDTA chelating. The binding of 3C7 to platelets increased significantly when platelets were activated by ADP/thrombin or occupied by RGDS peptides, fibrinogen, or PAC-1, suggesting that 3C7 was an anti-ligand induced binding site antibody. The antibody failed to bind to the CHO cells expressing another alpha IIb beta3 mutant (beta3Y178A) suggesting that the Cys177 Cys184 loop of beta3 was likely the epitope for 3C7. 3C7 inhibited platelet aggregation, which was initiated by ADP or thrombin in a dose-dependent manner (IC50s of 5.6 and 0.05 microg/ml, respectively). The antibody also inhibited platelet adhesion to immobilized fibrinogen but not to fibronectin or collagen. These findings suggested that 3C7 was a potent antagonist of integrin alpha IIb beta3 and a potential anti-thrombotic agent. PMID- 15890655 TI - Cholesterol-dependent lipid assemblies regulate the activity of the ecto nucleotidase CD39. AB - CD39 (ecto-nucleoside triphosphate diphosphohydrolase-1; E-NTPDase1) is a plasma membrane ecto-enzyme that regulates purinergic receptor signaling by controlling the levels of extracellular nucleotides. In blood vessels this enzyme exhibits a thromboregulatory role through the control of platelet aggregation. CD39 is localized in caveolae, which are plasma membrane invaginations with distinct lipid composition, similar to dynamic lipid microdomains, called rafts. Cholesterol is enriched together with sphingolipids in both rafts and caveolae, as well as in other specialized domains of the membrane, and plays a key role in their function. Here, we examine the potential role of cholesterol-enriched domains in CD39 function. Using polarized Madin-Darby canine kidney (MDCK) cells and caveolin-1 gene-disrupted mice, we show that caveolae are not essential either for the enzymatic activity of CD39 or for its targeting to plasma membrane. On the other hand, flotation experiments using detergent-free or detergent-based approaches indicate that CD39 associates, at least in part, with distinct lipid assemblies. In the apical membrane of MDCK cells, which lacks caveolae, CD39 is localized in microvilli, which are also cholesterol and raft dependent membrane domains. Interfering with cholesterol levels using drugs that either deplete or sequester membrane cholesterol results in a strong inhibition of the enzymatic and anti-platelet activity of CD39. The effects of cholesterol depletion are completely reversed by replenishment of membranes with pure cholesterol, but not by cholestenone. These data suggest a functional link between the localization of CD39 in cholesterol-rich domains of the membrane and its role in thromboregulation. PMID- 15890656 TI - A variable residue in the pore of Kv1 channels is critical for the high affinity of blockers from sea anemones and scorpions. AB - Animal toxins are associated with well defined selectivity profiles; however the molecular basis for this property is not understood. To address this issue we refined our previous three-dimensional models of the complex between the sea anemone toxin BgK and the S5-S6 region of Kv1.1 (Gilquin, B., Racape, J., Wrisch, A., Visan, V., Lecoq, A., Grissmer, S., Menez, A., and Gasparini, S. (2002) J. Biol. Chem. 277, 37406-37413) using a docking procedure that scores and ranks the structures by comparing experimental and back-calculated values of coupling free energies DeltaDeltaGint obtained from double-mutant cycles. These models further highlight the interaction between residue 379 of Kv1.1 and the conserved dyad tyrosine residue of BgK. Because the nature of the residue at position 379 varies from one channel subtype to another, we explored how these natural mutations influence the sensitivity of Kv1 channel subtypes to BgK using binding and electrophysiology experiments. We demonstrated that mutations at this single position indeed suffice to abolish or enhance the sensitivity of Kv1 channels for BgK and other sea anemone and scorpion toxins. Altogether, our data suggest that the residue at position 379 of Kv1 channels controls the affinity of a number of blocking toxins. PMID- 15890657 TI - Characterization of four variant forms of human propionyl-CoA carboxylase expressed in Escherichia coli. AB - Propionyl-CoA carboxylase (PCC) is a biotin-dependent mitochondrial enzyme that catalyzes the conversion of propionyl-CoA to D-methylmalonyl-CoA. PCC consists of two heterologous subunits, alpha PCC and beta PCC, which are encoded by the nuclear PCCA and PCCB genes, respectively. Deficiency of PCC results in a metabolic disorder, propionic acidemia, which is sufficiently severe to cause neonatal death. We have purified three PCCs containing pathogenic mutations in the beta subunit (R165W, E168K, and R410W) and one PCCB polymorphism (A497V) to homogeneity to elucidate the potential structural and functional effects of these substitutions. We observed no significant difference in Km values for propionyl CoA between wild-type and the variant enzymes, which indicated that these substitutions had no effect on the affinity of the enzyme for this substrate. Furthermore, the kinetic studies indicated that mutation R410W was not involved in propionyl-CoA binding in contrast to a previous report. The three mutant PCCs had half the catalytic efficiency of wild-type PCC as judged by the kcat/Km ratios. No significant differences have been observed in molecular mass or secondary structure among these enzymes. However, the variant PCCs were less thermostable than the wild-type. Following incubation at 47 degrees C, blue native-PAGE revealed a lower oligomeric form (alpha2beta2) in the three mutants not detectable in wild-type and the polymorphism. Interestingly, the lower oligomeric form was also observed in the corresponding crude Escherichia coli extracts. Our biochemical data and the structural analysis using a beta PCC homology model indicate that the pathogenic nature of these mutations is more likely to be due to a lack of assembly rather than disruption of catalysis. The strong favorable effect of the co-expressed chaperone proteins on PCC folding, assembly, and activity suggest that propionic acidemia may be amenable to chaperone therapy. PMID- 15890658 TI - N-acylphosphatidylethanolamine-hydrolyzing phospholipase D is an important determinant of uterine anandamide levels during implantation. AB - Implantation requires reciprocal interaction between blastocysts and a receptive uterus. In mice, one important player in this dialogue involves endocannabinoid signaling via cannabinoid receptor CB1. Anandamide is an endogenous cannabinoid ligand, and its levels are spatiotemporally regulated in the uterus during early pregnancy, showing lower levels in the receptive uterus and at the implantation site. However, the mechanism by which differential uterine anandamide gradients are established under different pregnancy status is not clearly understood. Using multiple approaches, we show here that uterine anandamide levels conducive to implantation are primarily regulated by spatiotemporal expression of Nape-Pld, the gene encoding N-acylphosphatidylethanolamine-hydrolyzing phospholipase D that generates anandamide. The expression is well correlated with its activity and anandamide levels. This study is clinically relevant, since elevated anandamide levels in peripheral circulation are associated with spontaneous pregnancy failure in women. PMID- 15890659 TI - Determining the environment of the ligand binding pocket of the human angiotensin II type I (hAT1) receptor using the methionine proximity assay. AB - The peptide hormone angiotensin II (AngII) binds to the AT0 (angiotensin type 1) receptor within the transmembrane domains in an extended conformation, and its C terminal residue interacts with transmembrane domain VII at Phe-293/Asn-294. The molecular environment of this binding pocket remains to be elucidated. The preferential binding of benzophenone photolabels to methionine residues in the target structure has enabled us to design an experimental approach called the methionine proximity assay, which is based on systematic mutagenesis and photolabeling to determine the molecular environment of this binding pocket. A series of 44 transmembrane domain III, VI, and VII X --> Met mutants photolabeled either with 125I-[Sar1,p'-benzoyl-L-Phe8]AngII or with 125I-[Sar1,p''-methoxy-p' benzoyl-L-Phe8]AngII were purified and digested with cyanogen bromide. Several mutants produced digestion patterns different from that observed with wild type human AT1, indicating that they had a new receptor contact with position 8 of AngII. The following residues form this binding pocket: L112M and Y113M in transmembrane domain (TMD) III; F249M, W253M, H256M, and T260M in TMD VI; and F293M, N294M, N295M, C296M, and L297M in TMD VII. Homology modeling and incorporation of these contacts allowed us to develop an evidence-based molecular model of interactions with human AT1 that is very similar to the rhodopsin retinal interaction. PMID- 15890660 TI - Regulation of apoptosis signal-regulating kinase 1 (ASK1) by polyamine levels via protein phosphatase 5. AB - Recent evidence has implicated the protein phosphatase PP5 in a variety of signaling pathways. Whereas several proteins have been identified that interact with PP5 and regulate its activity, a possibility of its regulation by second messengers remains speculative. Activation of PP5 in vitro by polyunsaturated fatty acids (e.g. arachidonic acid) and fatty acyl-CoA esters (e.g. arachidonoyl CoA) has been reported. We report here that PP5 is strongly inhibited by micromolar concentrations of a natural polyamine spermine. This inhibition was observed both in assays with a low molecular weight substrate p-nitrophenyl phosphate as well as phosphocasein and apoptosis signal-regulating kinase 1 (ASK1), thought to be a physiological substrate of PP5. Furthermore, a decrease in polyamine levels in COS-7 cells induced by alpha-difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase, led to accelerated dephosphorylation of oxidative stress-activated ASK1. This effect was suppressed by okadaic acid and by siRNA-mediated PP5 depletion, indicating that the effect of polyamine levels on ASK1 dephosphorylation was mediated by PP5. In line with the decreased ASK1 activation, polyamine depletion in COS-7 cells abrogated oxidative stress-induced activation of caspase-3, which executes ASK1-induced apoptosis, as well as caspase-3 activation induced by ASK1 overexpression, but had no effect on basal caspase-3 activity. These results implicate polyamines, emerging intracellular signaling molecules, as potential physiological regulators of PP5. Our findings also suggest a novel mechanism of the anti-apoptotic action of a decrease in polyamine levels via de-inhibition of PP5 and accelerated dephosphorylation and deactivation of ASK1. PMID- 15890661 TI - Surfactant protein A is a principal and oxidation-sensitive microbial permeabilizing factor in the alveolar lining fluid. AB - We have reported that surfactant protein A kills some Gram-negative organisms by increasing membrane permeability. In this study, we investigated the physiologic importance of this activity and the effect of oxidative stress on the antimicrobial functions of SP-A in vitro and in vivo. Concentrated bronchoalveolar lavage fluids from SP-A+/+ mice increased the permeability of the Escherichia coli K12 cell membrane to a greater extent than lavage from SP-A-/- animals. Similarly, calcium-dependent surfactant-binding proteins of SP-A+/+ mice increased membrane permeability more than those from SP-A-/- mice and produced greater zonal killing of agar-embedded bacteria in a radial diffusion assay. Exposure of human SP-A to copper-initiated surfactant phospholipid peroxidation or to free radicals generated by human neutrophils in vitro increased the level of SP-A-associated carbonyl moieties and blocked the permeabilizing function of the protein. We also found that exposure of mice to 90% O2 for 4 days, sufficient to lead to consumption of glutathione, oxidation of protein thiols, and accumulation of airspace protein-associated carbonyl moieties, blocked the permeabilizing activity of lavage fluid from SP-A+/+ mice. We conclude that SP-A is a major microbial permeablizing factor in lavage fluid and that oxidative stress inhibits the antibacterial activity of SP-A by a mechanism that includes oxidative modification and functional inactivation of the protein. PMID- 15890662 TI - Death in the dirt. PMID- 15890663 TI - Heart, heal thyself. PMID- 15890664 TI - Keep time, stay healthy. AB - Recent studies reveal that sleep loss in humans leads to metabolic disorders. A new study reports the development of metabolic syndrome in mice with altered circadian timing brought about by a mutation in a gene that functions in the biological clock. An intriguing and unanswered question is the relation between a healthy biological clock and normal appetite and weight regulation. PMID- 15890665 TI - Lower dose capecitabine has a more favorable therapeutic index in metastatic breast cancer: retrospective analysis of patients treated at M. D. Anderson Cancer Center and a review of capecitabine toxicity in the literature. AB - BACKGROUND: Capecitabine is active against anthracycline- and taxane-pretreated metastatic breast cancer. Post-marketing use of capecitabine at the FDA-approved dose (2500 mg/m2/day) leads to unacceptable toxicity in many patients. Dose reductions anecdotally improve tolerability without compromising efficacy. This retrospective analysis was designed to verify these anecdotal reports. PATIENTS AND METHODS: We retrospectively reviewed the records of 141 consecutive patients with metastatic breast cancer identified from pharmacy records as receiving capecitabine outside of a clinical trial between May 1998 and February 1999. Responses were defined as clinical improvement (ID), stabilization of disease (SD) for 6 weeks or longer, or progression (PD). Patients were grouped according to the starting dose level of capecitabine: A=2500+/-5% (dose range 2385-2560) mg/m2/day; B=2250+/-5% (range 2130-2350) mg/m2/day; C < or = 2000+5% (range 1000 2100) mg/m2/day. We also reviewed the safety profile of capecitabine at these doses and performed a safety review of capecitabine in phase II and III metastatic breast and colorectal cancer trials. RESULTS: Clinical data were available for 113 patients (105 for response, 106 for toxicity). The median age was 52.5 years and the mean number of prior metastatic chemotherapy regimens was 2 (range 0-7). The mean capecitabine starting dose was 2220 mg/m2/day and the median number of cycles administered was 4 (range 1-19). The mean tolerated dose was 2040 mg/m2/day (range 960-2670). Grade 3/4 toxic effects at dose levels A, B and C, respectively, included palmar-plantar erythrodysesthesia (33%, 63%, 20%), diarrhea (13%, 12%, 3%), stomatitis (8%, 0%, 3%), and nausea/vomiting (4%, 6%, 5%). Forty per cent of all patients required capecitabine dose reductions; fewer patients treated with 2000 mg/m2/day required dose modification (28%). Five per cent of the patients required discontinuation of capecitabine owing to toxicity. Patients started at the lowest doses of capecitabine did not have poorer response rates or shorter time to progression. CONCLUSIONS: This retrospective analysis supports a starting dose of 2000 mg/m2/day because of its superior therapeutic index; however, patients may still have toxic effects and individualization of dosing is necessary. A phase III, multicenter, randomized study to establish the safety and efficacy of different doses of capecitabine is urgently needed. PMID- 15890666 TI - Sentinel node biopsy is reliable in early-stage cervical cancer but not in locally advanced disease. AB - BACKGROUND: Sentinel lymph node (SN) biopsy based on dual labeling with blue dye and radiocolloid can reliably determine lymph node status in early-stage cervical cancer, but few data are available on its accuracy in more advanced disease. We examined the influence of tumor stage on the accuracy of SN biopsy in patients with cervical cancer. METHODS: Between July 2001 and June 2004, 33 patients (mean age 52 years) with early-stage or locally advanced cervical cancer underwent laparoscopic SN biopsy based on dual labeling with patent blue and radiocolloid. Patients with early-stage cervical cancer (stages IA and IB1, 23 patients) underwent complete laparoscopic pelvic lymphadenectomy after the SN procedure. Patients with locally advanced cervical cancer (stage IB2, IIA or IIB, 10 patients) underwent laparoscopic pelvic and para-aortic lymphadenectomy after SN biopsy and prior neoadjuvant concomitant chemoradiotherapy. The SN identification rates and false-negative rates of patients with early-stage and locally advanced disease were compared. RESULTS: The mean numbers of SNs identified per patient with early-stage and locally advanced cervical cancer were 2.3 (range 0-4) and 1.9 (range 0-4), respectively. SNs were identified in 86.9% (20/23) of patients with early-stage disease and in 80% (8/10) of patients with locally advanced disease. When analyzed according to the side of dissection, the identification rate was lower, especially in the patients with locally advanced disease (55% compared with 67.4%). The false-negative rate per patient was zero in early-stage disease and 20% (1/5) in locally advanced disease (no significant difference). When the side of dissection was taken into account, the false-negative rate improved to 42.9% (3/7) in patients with locally advanced disease and remained at zero in early-stage disease (P=0.038). Isolated blue dye was taken up in 53.3% of SNs in patients with locally advanced disease, compared with only 6.4% in patients with early-stage disease. CONCLUSIONS: This study suggests that the SN biopsy technique with dual labeling is less accurate in locally advanced cervical cancer than in early-stage cervical cancer. PMID- 15890667 TI - Metachronous gastric MALT lymphoma and early gastric cancer: is residual lymphoma a risk factor for the development of gastric carcinoma? AB - BACKGROUND: Helicobacter pylori plays a major role in the pathogenesis of primary gastric MALT lymphoma (GML) and gastric carcinoma. The occurrence of these two diseases metachronously in a same patient is a rare event. PATIENTS AND METHODS: Gastric biopsies and gastrectomy resection specimens of four patients who developed GML and early gastric cancer (EGC) were analysed by morphology, immunohistochemistry and molecular biology. RESULTS: Four patients (three males and one female; mean age 48 years) were diagnosed with GML. Helicobacter pylori infection was observed in three cases. Two patients had localized disease (stages IE and IIE, respectively) and were treated with H. pylori eradication therapy followed by an alkylating agent for one patient. Two patients had disseminated disease (stage IV), and were treated with an alkylating agent. Three cases were t(11;18) positive. All patients achieved initially complete lymphoma remission. Long-term endoscopic surveillance detected an EGC at the same location as the lymphoma in all patients at a mean time of 9.5 years (range 2.5-17 years) after lymphoma diagnosis. Gastrectomy specimens showed residual GML in all cases. CONCLUSION: Prolonged residual GML could constitute an additional risk factor for the development of gastric carcinoma. Long-term endoscopic surveillance is mandatory in patients treated conservatively for gastric MALT lymphoma. PMID- 15890668 TI - Irinotecan or oxaliplatin for first-line treatment of advanced colorectal cancer? PMID- 15890669 TI - Quality of cancer care. PMID- 15890670 TI - 1alpha,25-Dihydroxyvitamin D3 targets PTEN-dependent fibronectin expression to restore thyroid cancer cell adhesiveness. AB - We have previously reported that the hormonal form of 1alpha,25-dihydroxyvitamin D3 (1,25-VD3), and its noncalciomimetic analog EB1089, arrest the growth of human thyroid cancer cells by increasing the cell cycle inhibitor p27. In the present study, we investigated whether the tumor-suppressive effects of vitamin D (VD) compounds may also be mediated by mechanisms that govern cell adhesiveness. Both 1,25-VD3 and EB1089 increased cell adhesiveness, an effect that was accompanied by consistent increases in fibronectin (FN) expression. Introduction of small interfering RNA against FN resulted in down-regulation of FN expression and diminished cell adhesiveness to a collagen-type I matrix. To determine whether this action of 1,25-VD3 was mediated through the PTEN/phosphoinositol 3-kinase pathway, we examined whether this tumor suppressor protein/dual phosphatase can influence FN expression and consequently cell adhesiveness Overexpression of wild type PTEN induced FN expression as well as cell adhesiveness. In contrast, introduction of mutant forms of PTEN failed to induce FN and led to diminished cell adhesiveness. Conversely, small interfering RNA-mediated PTEN down regulation attenuated FN expression as well as cell adhesiveness. The attenuated FN expression was also associated with relative insensitivity to 1,25-VD3 growth suppressive action. Cells down-regulated for FN demonstrated a more aggressive growth pattern in xenografted mice and were also relatively insensitive to 1,25 VD3 treatment. Taken together, our findings highlight the significance of FN in modulating thyroid cancer cell adhesiveness and, at least in part, in mediating VD actions on neoplastic cell growth. PMID- 15890671 TI - Gonadotropin-releasing hormone induction of extracellular-signal regulated kinase is blocked by inhibition of calmodulin. AB - Our previous studies demonstrate that GnRH-induced ERK activation required influx of extracellular Ca2+ in alphaT3-1 and rat pituitary cells. In the present studies, we examined the hypothesis that calmodulin (Cam) plays a fundamental role in mediating the effects of Ca2+ on ERK activation. Cam inhibition using W7 was sufficient to block GnRH-induced reporter gene activity for the c-Fos, murine glycoprotein hormone alpha-subunit, and MAPK phosphatase (MKP)-2 promoters, all shown to require ERK activation. Inhibition of Cam (using a dominant negative) was sufficient to block GnRH-induced ERK but not c-Jun N-terminal kinase activity activation. The Cam-dependent protein kinase (CamK) II inhibitor KN62 did not recapitulate these findings. GnRH-induced phosphorylation of MAPK/ERK kinase 1 and c-Raf kinase was blocked by Cam inhibition, whereas activity of phospholipase C was unaffected, suggesting that Ca2+/Cam modulation of the ERK cascade potentially at the level of c-Raf kinase. Enrichment of Cam-interacting proteins using a Cam agarose column revealed that c-Raf kinase forms a complex with Cam. Reconstitution studies reveal that recombinant c-Raf kinase can associate directly with Cam in a Ca2+-dependent manner and this interaction is reduced in vitro by addition of W7. Cam was localized in lipid rafts consistent with the formation of a Ca2+-sensitive signaling platform including the GnRH receptor and c-Raf kinase. These data support the conclusion that Cam may have a critical role as a Ca2+ sensor in specifically linking Ca2+ flux with ERK activation within the GnRH signaling pathway. PMID- 15890672 TI - The 1,25(OH)2D3-regulated transcription factor MN1 stimulates vitamin D receptor mediated transcription and inhibits osteoblastic cell proliferation. AB - The vitamin D endocrine system is essential for maintaining mineral ion homeostasis and preserving bone density. The most bioactive form of vitamin D, 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] elicits its effects by binding to the vitamin D receptor (VDR) and regulating the transcription of target genes. In osteoblasts, the bone-forming cells of the skeleton, 1,25-(OH)2D3 regulates cell proliferation, differentiation, and mineralization of the extracellular matrix. Despite these well-characterized biological functions, relatively few 1,25 (OH)2D3 target genes have been described in osteoblasts. In this study, we characterize the regulation and function of MN1, a novel 1,25-(OH)2D3-induced gene in osteoblastic cells. MN1 is a nuclear protein first identified as a gene disrupted in some meningiomas and leukemias. Our studies demonstrate that MN1 preferentially stimulates VDR-mediated transcription through its ligand-binding domain and synergizes with the steroid receptor coactivator family of coactivators. Furthermore, forced expression of MN1 in osteoblastic cells results in a profound decrease in cell proliferation by slowing S-phase entry, suggesting that MN1 is an antiproliferative factor that may mediate 1,25-(OH)2D3-dependent inhibition of cell growth. Collectively, these data indicate that MN1 is a 1,25 (OH)2D3-induced VDR coactivator that also may have critical roles in modulating osteoblast proliferation. PMID- 15890673 TI - Selective expression of a dominant-negative form of peroxisome proliferator activated receptor in keratinocytes leads to impaired epidermal healing. AB - Many nuclear hormone receptors are involved in the regulation of skin homeostasis. However, their role in the epithelial compartment of the skin in stress situations, such as skin healing, has not been addressed yet. The healing of a skin wound after an injury involves three major cell types: immune cells, which are recruited to the wound bed; dermal fibroblasts; and epidermal and hair follicle keratinocytes. Our previous studies have revealed important but nonredundant roles of PPARalpha and beta/delta in the reparation of the skin after a mechanical injury in the adult mouse. However, the mesenchymal or epithelial cellular compartment in which PPARalpha and beta/delta play a role could not be determined in the null mice used, which have a germ line PPAR gene invalidation. In the present work, the role of PPARalpha was studied in keratinocytes, using transgenic mice that express a PPARalpha mutant with dominant-negative (dn) activity specifically in keratinocytes. This dn PPARalpha lacks the last 13 C terminus amino acids, binds to a PPARalpha agonist, but is unable to release the nuclear receptor corepressor and to recruit the coactivator p300. When selectively expressed in keratinocytes of transgenic mice, dn PPARalphaDelta13 causes a delay in the healing of skin wounds, accompanied by an exacerbated inflammation. This phenotype, which is similar to that observed in PPARalpha null mice, strongly suggests that during skin healing, PPARalpha is required in keratinocytes rather than in other cell types. PMID- 15890674 TI - Association of human follitropin (FSH) receptor with splicing variant of human lutropin/choriogonadotropin receptor negatively controls the expression of human FSH receptor. AB - A splice variant of human lutropin (LH)/choriogonadotropin (CG)-receptor [hLHR(exon 9)] that lacks exon 9 was previously cloned in the corpus luteum of a woman with a normal menstrual cycle. Supported by a detergent-soluble binding assay and a receptor biotinylation experiment, the receptor binding assay shows hLHR(exon 9) is neither expressed at the cell surface nor has the capability of binding to hCG. In addition, hLHR(exon 9) was confirmed in the endoplasmic reticulum (ER) by endoglycosidase H treatment. A coimmunoprecipitation experiment clearly showed that hLHR(exon 9) and constitutively inactivate mutant-LHRs, which stay in the ER, form an association with the human follitropin (FSH)-receptor (hFSHR). This suggests that in the presence of mutant-LHR, hFSHR, which is trapped in the ER and associated with hLHR(exon 9), is unable to come up to the plasma membrane. This phenomenon is specific among gonadotropin receptors because human TSH receptor failed to be coimmunoprecipitated. Furthermore, this receptor complex attenuated the hFSHR receptor protein level within the cells, which impaired cAMP production. To elucidate the mechanism underlying the decrease in hFSHR protein by this receptor complex, we performed a Percoll fractionation experiment, which indicated that the receptor complex drove hFSHR to the lysosome instead of the plasma membrane. These results reveal a novel mechanism of FSHR expression regulation. PMID- 15890675 TI - Haploinsufficiency of chicken ovalbumin upstream promoter transcription factor II in female reproduction. AB - The chicken ovalbumin upstream promoter transcription factor II, COUP-TFII, is a member of the orphan nuclear receptor transcription factor family. Genetic ablation of COUP-TFII results in early embryonic lethality and demonstrates that this gene is required for cardiac and vascular development. Expression of COUP TFII persists throughout postnatal life in various tissues including the female reproductive tract. However, the physiological function of COUP-TFII in female reproduction has not been extensively analyzed. Here, we provide phenotypic evidences that haploinsufficiency of COUP-TFII in mice demonstrates an important role of COUP-TFII for normal female reproduction. COUP-TFII +/- females show significantly reduced fecundity, irregular estrus cycles, delayed puberty, and retarded postnatal growth. Analysis of the reduced fertility revealed that although ovarian function was normal with respect to ovulation, the ovaries have reduced ability to synthesize progesterone in response to exogenous gonadotropins. This reduction is due to the reduction of the expression of steroidogenic enzymes important for progesterone synthesis and the reduction of vascularization in COUP-TFII heterozygotes. Analysis of uterine function demonstrated a reduced response to an experimentally induced decidual cell reaction indicating that the ability of the uterus to support embryo implantation was reduced. Taken together, our data show global impact of gene dosage effects of COUP-TFII on female postnatal life and indicates requirement of COUP-TFII in normal female reproduction, in particular for uterine endometrial functions during the peri-implantation period. PMID- 15890676 TI - Haploinsufficiency of cytochrome P450 17alpha-hydroxylase/17,20 lyase (CYP17) causes infertility in male mice. AB - Cytochrome P450 17alpha-hydroxylase/17,20-lyase (CYP17) is critical in determining cortisol and sex steroid biosynthesis. To investigate how CYP17 functions in vivo, we generated mice with a targeted deletion of CYP17. Although in chimeric mice Leydig cell CYP17 mRNA and intratesticular and circulating testosterone levels were dramatically reduced (80%), the remaining testosterone was sufficient to support spermatogenesis as evidenced by the generation of phenotypical black C57BL/6 mice. However, male chimeras consistently failed to generate heterozygous CYP17 mice and after five matings chimeric mice stopped mating indicating a change in sexual behavior. These results suggested that CYP17 deletion caused a primary phenotype (infertility), probably not due to the anticipated androgen imbalance and a secondary phenotype (change in sexual behavior) due to the androgen imbalance. Surprisingly, CYP17 mRNA was found in mature sperm, and serial analysis of gene expression identified CYP17 mRNA in other testicular germ cells. CYP17 mRNA levels were directly related to percent chimerism. Moreover, more than 50% of the sperm from high-percentage chimeric mice were morphologically abnormal, and half of them failed the swim test. Furthermore, 60% of swimming abnormal sperm was devoid of CYP17. These results suggest that CYP17, in addition to its role in steroidogenesis and androgen formation, is present in germ cells where it is essential for sperm function, and deletion of one allele prevents genetic transmission of mutant and wild-type alleles causing infertility followed by change in sexual behavior due to androgen imbalance. PMID- 15890677 TI - The coactivator p300 directly acetylates the forkhead transcription factor Foxo1 and stimulates Foxo1-induced transcription. AB - The FOXO (Forkhead box class O) subgroup of forkhead transcription factors controls the expression of many genes involved in fundamental cellular processes. Until recently, studies conducted on posttranslational modifications of Forkhead proteins were restricted to their phosphorylation. In this report, we show that the coactivator p300 directly acetylates lysines in the carboxyl-terminal region of Foxo1 in vivo and in vitro, and potently stimulates Foxo1-induced transcription of IGF-binding protein-1 in transient transfection experiments. The intrinsic acetyltransferase activity of p300 is required for both activities. Our results suggest that acetylation of Foxo1 by p300 is responsible, at least in part, for its increased transactivation potency, although acetylation of histones cannot be excluded. Insulin, the major negative regulator of Foxo1-stimulated transcription, potently enhances p300 acetylation of Foxo1. Three consensus protein kinase B/Akt phosphorylation sites whose phosphorylation is stimulated by insulin are required for insulin-induced acetylation of Foxo1. In contrast to its importance in regulating the transcriptional activity of Foxo1 in the absence of insulin, acetylation plays only a minor role compared with phosphorylation in insulin inhibition of Foxo1 transcriptional activity. PMID- 15890678 TI - Upper limb disability in HAVS cases--how does it relate to the neurosensory or vascular elements of HAVS? AB - AIMS: Hand-arm vibration syndrome (HAVS) consists of vascular, neurosensory and musculoskeletal components, characterized by symptoms that include Raynaud's phenomenon, tingling and numbness in the hands. However, there has been little published data on the effects of HAVS on the capability to carry out normal daily tasks. We have investigated the application of the widely-used disability, arm, shoulder and hand (DASH) disability questionnaire that reflects functionality problems in the upper extremities, as well as symptoms, in a HAVS cohort. METHODS: The cohort consisted of 118 males who, as a part of their health surveillance, had been referred for further assessment and Stockholm workshop staging. This process involved medical interview, physical examination and quantitative tests covering neurosensory function, manual dexterity and handgrip strength. RESULTS: The relationship between DASH outcome metric and a combination of quantitative tests reflecting a range of abnormalities found in HAVS, supports the validity of this questionnaire in HAVS studies. The data suggest that HAVS cases have a greater level of upper extremity disability compared with a general population. The study confirms that disability in HAVS is very largely related to sensorineural Stockholm workshop staging, rather than vascular staging. Any influence of vibration-induced Raynaud's phenomenon on upper extremity disability is related to the frequency of blanching attacks rather than their extent across the digits. CONCLUSION: This study strengthens the importance of identifying and preventing the exacerbation of the neurosenory component of HAVS, that unlike the blanching attacks of the vascular component does not have such an obvious pathognomic signal. PMID- 15890679 TI - PeMADS6, a GLOBOSA/PISTILLATA-like gene in Phalaenopsis equestris involved in petaloid formation, and correlated with flower longevity and ovary development. AB - In this study, we isolated and characterized the function of a GLOBOSA/PISTILLATA like gene, PeMADS6, from a native Phalaenopsis species, P. equestris. Southern blot analysis showed PeMADS6 as a single copy in the Phalaenopsis genome. Results of the determination of temporal and spatial expression showed that PeMADS6 was expressed and thus participated in the development of the sepals, petals, labellum and column in Phalaenopsis. Further confirmation of the expression pattern of PeMADS6 was carried out with in situ hybridization. Repressed expression of PeMADS6 in the orchid ovary was found to be pollination regulated, which suggests that the gene may have an inhibitory effect on the development of the ovary or ovule. In addition, auxin acted as the candidate signal to regulate the repression of PeMADS6 expression in the ovary. Furthermore, the flowers of transgenic Arabidopsis plants ectopically overexpressing PeMADS6 showed the morphology of petaloid sepals, with a 3- to 4-fold increase in flower longevity. Concomitantly, delayed fruit maturation was also observed in the transgenic Arabidopsis, which is consistent with the inhibitory effect of PeMADS6 on the development of the ovary. Thus, as a B-function gene, PeMADS6, not only specifies floral organ identity but has functions in flower longevity and ovary development in orchids. PMID- 15890680 TI - Modulation of fructokinase activity of potato (Solanum tuberosum) results in substantial shifts in tuber metabolism. AB - Potato plants (Solanum tuberosum L. cvs Desiree and Record) transformed with sense and antisense constructs of a cDNA encoding the potato fructokinase StFK1 exhibited altered transcription of this gene, altered amount of protein and altered enzyme activities. Measurement of the maximal catalytic activity of fructokinase revealed a 2-fold variation in leaf (from 90 to 180% of wild type activity) and either a 10- or 30-fold variation in tuber (from 10 or 30% to 300% in Record and Desiree, respectively) activity. The comparative effect of the antisense construct in leaf and tuber tissue suggests that this isoform is only a minor contributor to the total fructokinase activity in the leaf but the predominant isoform in the tuber. Antisense inhibition of the fructokinase resulted in a reduced tuber yield; however, its overexpression had no impact on this parameter. The modulation of fructokinase activity had few, consistent effects on carbohydrate levels, with the exception of a general increase in glucose content in the antisense lines, suggesting that this enzyme is not important for the control of starch synthesis. However, when metabolic fluxes were estimated, it became apparent that the transgenic lines display a marked shift in metabolism, with the rate of redistribution of radiolabel to sucrose markedly affected by the activity of fructokinase. These data suggest an important role for fructokinase, acting in concert with sucrose synthase, in maintaining a balance between sucrose synthesis and degradation by a mechanism independent of that controlled by the hexose phosphate-mediated activation of sucrose phosphate synthase. PMID- 15890681 TI - Cooperating pre-T-cell receptor and TCF-1-dependent signals ensure thymocyte survival. AB - Intrathymic T-cell maturation critically depends on the selective expansion of thymocytes expressing a functionally rearranged T-cell receptor (TCR) beta chain. In addition, TCR-independent signals also contribute to normal T-cell development. It is unclear whether and how signals from the 2 types of pathways are integrated. Here, we show that T-cell factor-1 (TCF-1), a nuclear effector of the canonical wingless/int (wnt)/catenin signaling pathway, ensures the survival of proliferating, pre-TCR(+) thymocytes. The survival of pre-TCR(+) thymocytes requires the presence of the N-terminal catenin-binding domain in TCF-1. This domain can bind the transcriptional coactivator beta-catenin and may also bind gamma-catenin (plakoglobin). However, in the absence of gamma-catenin, T-cell development is normal, supporting a role for beta-catenin. Signaling competent beta-catenin is present prior to and thus arises independently from pre-TCR signaling and does not substantially increase on pre-TCR signaling. In contrast, pre-TCR signaling significantly induces TCF-1 expression. This coincides with the activation of a wnt/catenin/TCF reporter transgene in vivo. Collectively, these data suggest that efficient TCF-dependent transcription requires that pre-TCR signaling induces TCF-1 expression, whereas wnt signals may provide the coactivator such as beta-catenin. The 2 pathways thus have to cooperate to ensure thymocyte survival at the pre-TCR stage. PMID- 15890682 TI - ADAMTS13 autoantibodies in patients with thrombotic microangiopathies and other immunomediated diseases. AB - Autoantibodies neutralizing human ADAMTS13 (a disintegrin-like and metalloproteinase with thrombospondin type 1 motif), the metalloprotease that physiologically cleaves von Willebrand factor, are a major cause of severe deficiency of the protease and of acquired thrombotic thrombocytopenic purpura (TTP). We evaluated prevalence of anti-ADAMTS13 antibodies in 59 patients with thrombotic microangiopathies (TMAs) and in 160 patients with immunologic or thrombocytopenic diseases different from TTP, using an enzyme-linked immunosorbent assay (ELISA). Immunoglobulin G (IgG) antibodies directed against ADAMTS13 were found in 97% of untreated patients with acute acquired TMA who had plasma levels of ADAMTS13 activity below 10%. The corresponding prevalence of IgM antibodies was 11%. In contrast, anti-ADAMTS13 antibodies of G or M isotypes were detected in 20% of patients with TMA with ADAMTS13 activity above 10%. The ELISA was more sensitive than the standard functional inhibitor assay for detecting antibodies against ADAMTS13. Patients with thrombocytopenia from various causes (n = 50), systemic lupus erythematosus (SLE; n = 40), and the antiphospholipid antibody syndrome (APS; n = 55) had prevalences of IgG antibodies of 8%, 13%, and 5% respectively, only slightly higher than the prevalence in 111 healthy donors (4%). A rather high prevalence of anti-ADAMTS13 IgM antibodies was found in patients with SLE and APS (18% each). The clinical significance of IgM antibodies in these groups is unclear. In conclusion, the ELISA method detected anti ADAMTS13 IgG antibodies in a very large proportion of patients with acquired TMA associated with severe ADAMTS13 deficiency, and was more sensitive than the inhibitor assay. PMID- 15890683 TI - How do stem cells find their way home? AB - Migration of hematopoietic stem cells through the blood, across the endothelial vasculature to different organs and to their bone marrow (BM) niches, requires active navigation, a process termed homing. Homing is a rapid process and is the first and essential step in clinical stem cell transplantation. Similarly, homing is required for seeding of the fetal BM by hematopoietic progenitors during development. Homing has physiological roles in adult BM homeostasis, which are amplified during stress-induced recruitment of leukocytes from the BM reservoir and during stem cell mobilization, as part of host defense and repair. Homing is thought to be a coordinated, multistep process, which involves signaling by stromal-derived factor 1 (SDF-1) and stem cell factor (SCF), activation of lymphocyte function-associated antigen 1 (LFA-1), very late antigen 4/5 (VLA-4/5) and CD44, cytoskeleton rearrangement, membrane type 1 (MT1)-matrix metalloproteinase (MMP) activation and secretion of MMP2/9. Rolling and firm adhesion of progenitors to endothelial cells in small marrow sinusoids under blood flow is followed by trans-endothelial migration across the physical endothelium/extracellular matrix (ECM) barrier. Stem cells finalize their homing uniquely, by selective access and anchorage to their specialized niches in the extravascular space of the endosteum region and in periarterial sites. This review is focused on mechanisms and key regulators of human stem cell homing to the BM in experimental animal models and clinical transplantation protocols. PMID- 15890684 TI - Conditional overexpression of transgenes in megakaryocytes and platelets in vivo. AB - Megakaryocyte (MK)-specific transgene expression has proved valuable in studying thrombotic and hemostatic processes. Constitutive expression of genes, however, could result in altered phenotypes due to compensatory mechanisms or lethality. To circumvent these limitations, we used the tetracycline/doxycycline (Tet)-off system to conditionally over-express genes in megakaryocytes and platelets in vivo. We generated 3 transactivator transgenic lines expressing the Tet transactivator element (tTA), under the control of the MK-specific platelet factor 4 promoter (PF4-tTA-VP16). Responder lines were simultaneously generated, each with a bidirectional minimal cytomegalovirus (CMV)-tTA responsive promoter driving prokaryotic beta-galactosidase gene, as a cellular reporter, and a gene of interest (in this case, the mitotic regulator Aurora-B). A transactivator founder line that strongly expressed PF4-driven tTA-viral protein 16 (VP16) was crossbred to a responder line. The homozygous double-transgenic mouse line exhibited doxycycline-dependent transgene overexpression in MKs and platelets. Using this line, platelets were conveniently indicated at sites of induced stress by beta-galactosidase staining. In addition, we confirmed our earlier report on effects of constitutive expression of Aurora-B, indicating a tight regulation at protein level and a modest effect on MK ploidy. Hence, we generated a new line, PF4-tTA-VP16, that is available for conditionally overexpressing genes of interest in the MK/platelet lineage in vivo. PMID- 15890685 TI - The use of AMD3100 plus G-CSF for autologous hematopoietic progenitor cell mobilization is superior to G-CSF alone. AB - Hematopoietic progenitor cells (HPCs) traffic to and are retained in the marrow through the trophic effects of the chemokine stromal cell-derived factor-1alpha (SDF-1alpha) binding to its receptor, CXC chemokine receptor 4 (CXCR4). AMD3100 reversibly inhibits SDF-1alpha/CXCR4 binding, and AMD3100 administration mobilizes CD34(+) cells into the circulation. We therefore tested the hypotheses that the combination of AMD3100 plus granulocyte colony-stimulating factor (G CSF) (hereafter A + G) would be superior to G-CSF alone (hereafter G) in mobilizing hematopoietic progenitor cells (HPCs) and that A + G-mobilized cells would engraft as well as G-mobilized cells. The primary objective was to determine whether patients mobilized more progenitor cells per unit of blood volume of apheresis after A + G administration versus G alone. Secondary objectives were to determine whether patients mobilized with A + G compared with G alone required fewer apheresis procedures to reach the target level at least 5 x 10(6) CD34(+) cells/kg for transplantation and to determine whether patients mobilized with A + G had at least a 90% success rate of autologous transplantation as assessed by neutrophil engraftment by day 21. Each patient served as his or her own control in a sequential mobilization design. All study objectives were met without significant toxicity. The results demonstrate that the combination of A + G is generally safe, effective, and superior to G alone for autologous HPC mobilization. PMID- 15890686 TI - Inhibition of APC anticoagulant activity on oxidized phospholipid by anti-{beta}2 glycoprotein I monoclonal antibodies. AB - Activated protein C (APC) anticoagulant activity and the ability to be inhibited by auto-antibodies associated with thrombosis are strongly augmented by the presence of phosphatidylethanolamine (PE) and phospholipid oxidation. beta(2) glycoprotein I (beta(2)-GPI) is a major antigen for antiphospholipid antibodies present in patients with the antiphospholipid syndrome. We therefore investigated whether anti-beta(2)-GPI monoclonal antibodies (mAbs) could inhibit APC with similar membrane specificity. Five mouse mAbs that reacted with different epitopes on beta(2)-GPI were examined. Each inhibited the PE-, phospholipid oxidation-dependent enhancement of APC anticoagulant activity and required antibody divalency. A chimeric APC that retains anticoagulant activity but is relatively unaffected by protein S, PE, or oxidation was not inhibited by the antibodies. In purified systems, anti-beta(2)-GPI mAb inhibition of factor Va inactivation was greater in the presence of protein S and required beta(2)-GPI. Surprisingly, although the mAbs did increase beta(2)-GPI affinity for membranes, PE and oxidation had little influence on the affinity of the beta(2)-GPI antibody complex for the membrane vesicles. We conclude that antibodies to beta(2)-GPI inhibit APC function specifically and contribute to a hypercoaguable state by disrupting specific protein-protein interactions induced by oxidation of PE containing membranes. PMID- 15890687 TI - Overexpression of sphingosine kinase 1 is an oncogenic event in erythroleukemic progression. AB - The erythroleukemia developed by spi-1/PU.1-transgenic mice is a model of multistage oncogenic process. Isolation of tumor cells representing discrete stages of leukemic progression enables the dissection of some of the critical events required for malignant transformation. To elucidate the molecular mechanisms of multistage leukemogenesis, we developed a microarray transcriptome analysis of nontumorigenic (HS1) and tumorigenic (HS2) proerythroblasts from spi 1-transgenic mice. The data show that transcriptional up-regulation of the sphingosine kinase gene (SPHK1) is a recurrent event associated with the tumorigenic phenotype of these transgenic proerythroblasts. SPHK1 is an enzyme of the metabolism of sphingolipids, which are essential in several biologic processes, including cell proliferation and apoptosis. HS1 erythroleukemic cells engineered to overexpress the SPHK1 protein exhibited growth proliferative advantage, increased clonogenicity, and resistance to apoptosis in reduced serum level by a mechanism involving activation of the extracellular signal-related kinases 1/2 (ERK1/2) and phosphatidylinositol 3-kinase (PI3K)/AKT pathways. In addition, SPHK1-overexpressing HS1 cells acquired tumorigenicity when engrafted in vivo. Finally, enforced expression of a dominant-negative mutant of SPHK1 in HS2 tumorigenic cells or treatment with a pharmacologic inhibitor reduced both cell growth and apoptosis resistance. Altogether, these data suggest that overexpression of the sphingosine kinase may represent an oncogenic event during the multistep progression of an erythroleukemia. PMID- 15890688 TI - Chimeric NK-receptor-bearing T cells mediate antitumor immunotherapy. AB - NKG2D is an activating cell-surface receptor expressed on natural killer (NK) cells and some T-cell subsets. Its ligands are primarily expressed on tumor cells. The aim of this study was to determine whether chimeric NK-receptor bearing T cells would directly kill tumor cells and lead to induction of host immunity against tumors. Chimeric NK receptors were produced by linking NKG2D or DNAX activating protein of 10 kDa (Dap10) to the cytoplasmic portion of the CD3zeta chain. Our results showed that chimeric (ch) NKG2D-bearing T cells responded to NKG2D-ligand-bearing tumor cells (RMA/Rae-1beta, EG7) but not to wild-type tumor cells (RMA). This response was dependent upon ligand expression on the target cells but not on expression of major histocompatibility complex (MHC) molecules, and the response could be blocked by anti-NKG2D antibodies. These T cells produced large amounts of T-helper 1 (Th1) cytokines and proinflammatory chemokines and killed ligand-expressing tumor cells. Adoptive transfer of chNKG2D-bearing T cells inhibited RMA/Rae-1beta tumor growth in vivo. Moreover, mice that had remained tumor-free were resistant to subsequent challenge with the wild-type RMA tumor cells, suggesting the generation of immunity against other tumor antigens. Taken together, our findings indicate that modification of T cells with chimeric NKG2D receptors represents a promising approach for immunotherapy against cancer. PMID- 15890689 TI - Malaria-specific transgenic CD4(+) T cells protect immunodeficient mice from lethal infection and demonstrate requirement for a protective threshold of antibody production for parasite clearance. AB - T cells are important in the immune response to malaria, both for their cytokines and their help for antibody production. To look at the relative importance of these roles, a T-cell receptor (TCR) transgenic mouse has been generated carrying a TCR specific for an epitope of the merozoite surface protein 1 (MSP-1) of the malaria parasite, Plasmodium chabaudi. In adoptive transfer experiments, malaria specific CD4(+) T cells expand and produce interferon gamma (IFN-gamma) early in infection, but the population contracts quickly despite prolonged persistence of the parasite. MSP-1-specific CD4(+) cells can protect immunodeficient mice from lethal infection; however, the parasite is only completely cleared in the presence of B cells showing that T helper cells are critical. Levels of malaria specific antibody and the speed of their production clearly correlate with the time of resolution of infection, indicating that a critical threshold of antibody production is required for parasite clearance. Furthermore, T cells specific for a shed portion of MSP-1 are able to provide help for antibody to the protective region, which remains bound to the infected erythrocyte, suggesting that MSP-1 has all of the components necessary for a good vaccine. PMID- 15890692 TI - Should parents accompany critically ill children during inter-hospital transport? AB - BACKGROUND: Parental accompaniment during inter-hospital transportation (retrieval) of critically ill children is not commonplace in the United Kingdom. METHODS: A three month pilot of parental accompaniment was undertaken in 2002 (143 retrievals), after which time the policy was adopted as standard practice. A follow up audit was performed in 2004 (136 retrievals). RESULTS: Findings were remarkably consistent between the two periods. Staff perceived little or no added stress during the majority of transfers (96% in 2002, 98% in 2004), and felt able to perform medical interventions without hindrance (98% in 2002, 100% in 2004). There was good agreement between medical and nursing staff regarding perception of stress and ability to perform interventions (phi statistic 0.57 to 1.00). Adverse events occurred during 11 (3.9%) retrievals; six of these involved a parent exclusively. Stress tended to be associated with adverse events or parental behaviour rather than disease acuity. Staff vetoed the offer of accompaniment on 11 occasions, for a variety of reasons. The majority of parents found the experience safe, beneficial, and perceived a reduction in stress as a result. These data may inform other retrieval services who are considering adopting a similar policy. PMID- 15890690 TI - c-Jun N-terminal kinase (JNK) is required for survival and proliferation of B lymphoma cells. AB - Several primary murine and human B lymphomas and cell lines were found to constitutively express high levels of the activated form of c-jun N-terminal kinase (JNK), a member of the mitogen-activated protein (MAP) kinase family. Proliferation of murine B lymphomas CH31, CH12.Lx, BKS-2, and WEHI-231 and the human B lymphomas BJAB, RAMOS, RAJI, OCI-Ly7, and OCI-Ly10 was strongly inhibited by SP600125, an anthrapyrazolone inhibitor of JNK, in a dose-dependent manner. The lymphoma cells underwent apoptosis and arrested at the G2/M phase of cell cycle. Furthermore, JNK-specific small interfering RNA (siRNA) inhibited the growth of both murine and human B lymphomas. Thus in the B-lymphoma model, JNK appears to have a unique prosurvival role. Survival signals provided by CD40 and interleukin-10 (IL-10) together reversed the growth inhibition induced by the JNK inhibitor. c-Myc protein levels were reduced in the presence of both SP600125 and JNK-specific siRNA, and CD40 ligation restored c-Myc levels. Moreover, Bcl-xL rescued WEHI-231 cells from apoptosis induced by the JNK inhibitor. The JNK inhibitor also reduced levels of early growth response gene-1 (Egr-1) protein, and overexpressing Egr-1 partially rescued lymphoma cells from apoptosis. Thus, JNK may act via c-Myc and Egr-1, which were shown to be important for B-lymphoma survival and growth. PMID- 15890693 TI - Prediction of vesicoureteral reflux after a first febrile urinary tract infection in children: validation of a clinical decision rule. AB - AIMS: To test the reproducibility of a highly sensitive clinical decision rule proposed to predict vesicoureteral reflux (VUR) after a first febrile urinary tract infection in children. This rule combines clinical (family history of uropathology, male gender, young age), biological (raised C reactive protein), and radiological (urinary tract dilation on renal ultrasound) predictors in a score, and provides 100% sensitivity. METHODS: A retrospective hospital based cohort study included all children, 1 month to 4 years old, with a first febrile urinary tract infection. The sensitivities and specificities of the rule at the two previously proposed score thresholds (< or =0 and < or =5) to predict respectively, all-grade or grade > or =3 VUR, were calculated. RESULTS: A total of 149 children were included. VUR prevalence was 25%. The rule yielded 100% sensitivity and 3% specificity for all-grade VUR, and 93% sensitivity and 13% specificity for grade > or =3 VUR. Some methodological weaknesses explain this lack of reproducibility. CONCLUSIONS: The reproducibility of the previously proposed decision rule was poor and its potential contribution to clinical management of children with febrile urinary tract infection seems to be modest. PMID- 15890694 TI - Heterotaxia syndrome: the role of screening for intestinal rotation abnormalities. AB - BACKGROUND: Heterotaxia syndrome involves multiple anomalies, including cardiac malformations and intestinal rotation abnormalities. Most authors recommend routine radiological evaluation, with laparotomy and Ladd procedure if a rotation abnormality is found. AIMS: To determine if routine radiological screening is necessary, and if there is a group of children that can safely be managed expectantly. METHODS: Retrospective chart review of all children with heterotaxia syndrome from 1968 to 2002. RESULTS: Complete data were available for 177 patients. Twenty five (14%) had neonatal gastrointestinal symptoms (feeding intolerance, vomiting). Eleven of these had gastrointestinal contrast studies, of which seven were abnormal and led to surgery. Of the 152 asymptomatic neonates, nine had radiological screening and six of these were abnormal. Only one was thought to have a narrow based mesentery, but did not undergo surgery due to cardiac disease. There were no intestinal complications on follow up in this group. The other 143 asymptomatic children did not undergo radiological screening and were closely followed. Four subsequently developed gastrointestinal symptoms and had contrast studies; only one of these had malrotation and underwent a Ladd procedure. Of the remaining 139 patients who remained asymptomatic, 60 (43%) died of cardiac disease and none developed intestinal symptoms or complications related to malrotation on follow up. CONCLUSION: Asymptomatic children with heterotaxia syndrome have a low risk of adverse outcome related to intestinal rotation abnormalities. Routine screening may not be necessary as long as close follow up is done, and prompt investigation is performed for those that develop gastrointestinal symptomatology. PMID- 15890695 TI - What is the long term outcome for children who fail to thrive? A systematic review. AB - AIMS: To ascertain the long term outcomes in children diagnosed as having failure to thrive (FTT). METHODS: Systematic review of cohort studies. Medline, Psychinfo, Embase, Cinahl, Web of Science, Cochrane, and DARE databases were searched for potentially relevant studies. INCLUSION CRITERIA: cohort studies or randomised controlled trials in children <2 years old with failure to thrive defined as weight <10th centile or lower centile and/or weight velocity <10th centile, with growth, development, or behaviour measured at 3 years of age or older. RESULTS: Thirteen studies met the inclusion criteria; eight included a comparison group, of which five included children identified in community settings. Two were randomised controlled trials. Attrition rates were 10-30%. Data from population based studies with comparison groups and which reported comparable outcomes in an appropriate form were pooled in a random effects meta analysis. Four studies report IQ scores at follow up and the pooled standardised mean difference was -0.22 (95% CI -0.41 to -0.03). Two studies reported growth data as standard deviation scores. Their pooled weighted mean difference for weight was -1.24 SDS (95% CI -2.00 to -0.48), and for height -0.87 SDS (95% CI 1.47 to -0.28). No studies corrected for parental height, but two reported that parents of index children were shorter. CONCLUSIONS: The IQ difference (equivalent to approximately 3 IQ points) is of questionable clinical significance. The height and weight differences are larger, but few children were below the 3rd centile at follow up. It is unclear to what extent observed differences reflect causal relations or confounding due to other variables. In the light of these results the aggressive approach to identification and management of failure to thrive needs reassessing. PMID- 15890696 TI - Circumcision for the prevention of urinary tract infection in boys: a systematic review of randomised trials and observational studies. AB - OBJECTIVE: To undertake a meta-analysis of published data on the effect of circumcision on the risk of urinary tract infection (UTI) in boys. DATA SOURCES: Randomised controlled trials and observational studies comparing the frequency of UTI in circumcised and uncircumcised boys were identified from the Cochrane controlled trials register, MEDLINE, EMBASE, reference lists of retrieved articles, and contact with known investigators. METHODS: Two of the authors independently assessed study quality using the guidelines provided by the MOOSE statement for quality of observational studies. A random effects model was used to estimate a summary odds ratio (OR) with 95% confidence intervals (CI). RESULTS: Data on 402,908 children were identified from 12 studies (one randomised controlled trial, four cohort studies, and seven case-control studies). Circumcision was associated with a significantly reduced risk of UTI (OR = 0.13; 95% CI, 0.08 to 0.20; p<0.001) with the same odds ratio (0.13) for all three types of study design. CONCLUSIONS: Circumcision reduces the risk of UTI. Given a risk in normal boys of about 1%, the number-needed-to-treat to prevent one UTI is 111. In boys with recurrent UTI or high grade vesicoureteric reflux, the risk of UTI recurrence is 10% and 30% and the numbers-needed-to-treat are 11 and 4, respectively. Haemorrhage and infection are the commonest complications of circumcision, occurring at rate of about 2%. Assuming equal utility of benefits and harms, net clinical benefit is likely only in boys at high risk of UTI. PMID- 15890697 TI - Differential responses to CO2 and sympathetic stimulation in the cerebral and femoral circulations in humans. AB - The relative importance of CO2 and sympathetic stimulation in the regulation of cerebral and peripheral vasculatures has not been previously studied in humans. We investigated the effect of sympathetic activation, produced by isometric handgrip (HG) exercise, on cerebral and femoral vasculatures during periods of isocapnia and hypercapnia. In 14 healthy males (28.1 +/- 3.7 (mean +/- S.D.) years), we measured flow velocity (VP; transcranial Doppler ultrasound) in the middle cerebral artery during euoxic isocapnia (ISO, +1 mmHg above rest) and two levels of euoxic hypercapnia (HC5, end-tidal P(CO(2)), P(ET,CO2), = +5 mmHg above ISO; HC10, P(ET,CO2) = +10 above ISO). Each P(ET,CO2) level was maintained for 10 min using the dynamic end-tidal forcing technique, during which increases in sympathetic activity were elicited by a 2-min HG at 30% of maximal voluntary contraction. Femoral blood flow (FBF; Doppler ultrasound), muscle sympathetic nerve activity (MSNA; microneurography) and mean arterial pressure (MAP; Portapres) were also measured. Hypercapnia increased VP and FBF by 5.0 and 0.6% mmHg-1, respectively, and MSNA by 20-220%. Isometric HG increased MSNA by 50% and MAP by 20%, with no differences between ISO, HC5 and HC10. During the ISO HG there was an increase in cerebral vascular resistance (CVR; 20 +/- 11%), while VP remained unchanged. During HC5 and HC10 HG, VP increased (13% and 14%, respectively), but CVR was unchanged. In contrast, HG-induced sympathetic stimulation increased femoral vascular resistance (FVR) during ISO, HC5 and HC10 (17-41%), while there was a general decrease in FBF below ISO. The HG-induced increases in MSNA were associated with increases in FVR in all conditions (r = 0.76-0.87), whereas increases in MSNA were associated with increases in CVR only during ISO (r = 0.91). In summary, in the absence of hypercapnia, HG exercise caused cerebral vasoconstriction, myogenically and/or neurally, which was reflected by increases in CVR and a maintained VP. In contrast, HG increased FVR during conditions of ISO, HC5 and HC10. Therefore, the cerebral circulation is more responsive to alterations in PCO2, and less responsive to sympathetic stimulation than the femoral circulation. PMID- 15890698 TI - ERK1/2 and p38 regulate trophoblasts differentiation in human term placenta. AB - Mitogen-activated protein kinases (MAPKs) control many cellular events from complex programmes, such as embryogenesis, cell differentiation and proliferation, and cell death, to short-term changes required for homeostasis and acute hormonal responses. However, little is known about expression and activation of classical MAPKs, extracellular signal-regulated kinase1/2 (ERK1/2) and p38 in human placenta. Therefore, we examined the expression of ERK1/2 and p38 in trophoblasts from human term placenta, and their implication in differentiation. In vitro, freshly isolated cytotrophoblast cells, cultivated in 10% fetal bovine serum (FBS), spontaneously aggregate and fuse to form multinucleated cells that phenotypically resemble mature syncytiotrophoblasts, that concomitantly produce human chorionic gonadotropin (hCG) and human placental lactogen (hPL). This study shows that the level of ERK1/2 and p38 decreases with increasing days of culture, to reach an undetectable level after 5 days of culture. Moreover, pretreatment of cells with an ERK1/2-specific inhibitor (PD98059) and/or a p38-specific inhibitor (SB203580) suppressed trophoblast differentiation. Our results also demonstrate that the p38 pathway is highly solicited as compared to the ERK1/2 pathway in the differentiation process. Furthermore, ERK1/2 and p38 are rapidly activated upon addition of FBS, but the activation of p38 is delayed compared to that of ERK1/2. In summary, this study showed that ERK1/2 and p38 pathways are essential to mediate initiation of trophoblast differentiation. PMID- 15890699 TI - Action potential fidelity during normal and epileptiform activity in paired soma axon recordings from rat hippocampus. AB - Although action potential initiation and propagation are fundamental to nervous system function, there are few direct electrophysiological observations of propagating action potentials in small unmyelinated fibres, such as the axons within mammalian hippocampus. To circumvent limitations of previous studies that relied on extracellular stimulation, we performed dual recordings: whole-cell recordings from hippocampal CA3 pyramidal cell somas and extracellular recordings from their axons, up to 800 micro m away. During brief spike trains under normal conditions, axonal spikes were more resistant to amplitude reduction than somatic spikes. Axonal amplitude depression was greatest at the axon initial segment < 150 microm from the soma, and initiation occurred approximately 75 microm from the soma. Although prior studies, which failed to verify spike initiation, suggested substantial axonal depression during seizure-associated extracellular K+([K+]o) rises, we found that 8 mm [K+]o caused relatively small decreases in axonal spike amplitude during brief spike trains. However, during sustained, epileptiform spiking induced in 8 mm [K+]o, axonal waveforms decreased significantly in peak amplitude. During epileptiform spiking, bursts of two or more action potentials > 20 Hz failed to propagate in most cases. In normal [K+]o at 25 and 32 degrees C, spiking superimposed on sustained somatic depolarization, but not spiking alone, produced similar axonal changes as the epileptiform activity. These results highlight the likely importance of steady-state inactivation of axonal channels in maintaining action potential fidelity. Such changes in axonal propagation properties could encode information and/or serve as an endogenous brake on seizure propagation. PMID- 15890700 TI - Cytosolic Cl- ions in the regulation of secretory and endocytotic activity in melanotrophs from mouse pituitary tissue slices. AB - Cl- ions are known regulators of Ca2+ -dependent secretory activity in many endocrine cells. The suggested mechanisms of Cl- action involve the modulation of GTP-binding proteins, voltage-activated calcium channels or maturation of secretory vesicles. We examined the role of cytosolic Cl- ([Cl-]i) and Cl- currents in the regulation of secretory activity in mouse melanotrophs from fresh pituitary tissue slices by using the whole-cell patch-clamp. We confirmed that elevated [Cl-]i augments Ca2- -dependent exocytosis and showed that Cl- acts on secretory vesicle maturation. The latter process was abolished by a V-type H- ATPase blocker (bafilomycin), intracellular 4,4'-diisothiocyanatostilbene-2,2' disulphonic acid (DIDS), a Cl- channel blocker, and tolbutamide, a sulphonylurea implicated in secretory vesicle maturation. In a small subset of cells, block of plasmalemmal Cl- current by DIDS reversibly enhanced endocytosis. The direct activation of G-proteins by GTP-gamma-S, a non-hydrolysable GTP analogue, did not restore the impaired secretion observed in low [Cl-]i conditions. The amplitude of voltage-activated calcium currents was unaffected by the [Cl-]i. Furthermore, two Cl- -permeable channels, calcium-activated Cl- channels and GABAA receptors, appeared as major regulators of intracellular Cl- homeostasis. In conclusion, the predominant underlying mechanism of Cl- action is mediated by intracellular Cl- fluxes during vesicle maturation, rather than activation of G-proteins or modulation of voltage-activated Ca2+channels. PMID- 15890701 TI - Comparative gene expression profile of mouse carotid body and adrenal medulla under physiological hypoxia. AB - The carotid body (CB) is an arterial chemoreceptor, bearing specialized type I cells that respond to hypoxia by closing specific K+ channels and releasing neurotransmitters to activate sensory axons. Despite having detailed information on the electrical and neurochemical changes triggered by hypoxia in CB, the knowledge of the molecular components involved in the signalling cascade of the hypoxic response is fragmentary. This study analyses the mouse CB transcriptional changes in response to low PO2 by hybridization to oligonucleotide microarrays. The transcripts were obtained from whole CBs after mice were exposed to either normoxia (21% O2), or physiological hypoxia (10% O2) for 24 h. The CB transcriptional profiles obtained under these environmental conditions were subtracted from the profile of control non-chemoreceptor adrenal medulla extracted from the same animals. Given the common developmental origin of these two organs, they share many properties but differ specifically in their response to O2. Our analysis revealed 751 probe sets regulated specifically in CB under hypoxia (388 up-regulated and 363 down-regulated). These results were corroborated by assessing the transcriptional changes of selected genes under physiological hypoxia with quantitative RT-PCR. Our microarray experiments revealed a number of CB-expressed genes (e.g. TH, ferritin and triosephosphate isomerase) that were known to change their expression under hypoxia. However, we also found novel genes that consistently changed their expression under physiological hypoxia. Among them, a group of ion channels show specific regulation in CB: the potassium channels Kir6.1 and Kcnn4 are up-regulated, while the modulatory subunit Kcnab1 is down-regulated by low PO2 levels. PMID- 15890702 TI - Extension and magnitude of denervation in skeletal muscle from ageing mice. AB - In this work we hypothesized that denervation in flexor digitorum brevis (FDB) muscle from ageing mice is more extensive than predicted by standard functional and structural assays used in the past. In addition, we asked whether denervation is a fully or partially developed process. Despite the reported alteration in skeletal muscle innervation, the quantification of the extension and magnitude of denervation in ageing rodents has remained elusive. To address these two questions we utilized a combination of electrophysiological and immunohistochemical assays directed to detecting the expression of tetrodotoxin (TTX)-resistant sodium channels (Na(v)1.5) in FDB muscles from young-adult and senescent mice. Sodium current density measured with the macropatch cell-attached technique did not show significant differences between FDB fibres from young and old mice. The TTX dose-response curve, using the whole cell voltage-clamp technique, showed three populations of fibres in senescent mice, one similar to fibres from young mice (TTX sensitive), another one similar to fibres from experimentally denervated muscle (TTX resistant), and a third group intermediate between these two. Partially and fully denervated fibres added up to approximately 50% of the total number of fibres tested, a number that concurs with the percentage of fibres positive for the Na(v)1.5 channel by specific immunostaining. PMID- 15890703 TI - Expression and function of dipeptidyl-aminopeptidase-like protein 6 as a putative beta-subunit of human cardiac transient outward current encoded by Kv4.3. AB - Dipeptidyl-aminopeptidase-like protein 6 (DPPX) was recently shown in the brain to modulate the kinetics of transient A-type currents by accelerating inactivation and recovery from inactivation. Since the kinetics of human cardiac transient outward current (I(to)) are not mimicked by coexpression of the alpha subunit Kv4.3 with its known beta-subunit KChIP2, we have tested the hypothesis that DPPX may serve as an additional beta-subunit in the human heart. With quantitative real-time RT-PCR strong mRNA expression of DPPX was detected in human ventricles and was verified at the protein level in human but not in rat heart by a DPPX-specific antibody. Co-expression of DPPX with Kv4.3 in Chinese hamster ovary cells produced I(to)-like currents, but compared with expression of KChIP2a and Kv4.3, the time constant of inactivation was faster, the potential of half-maximum steady-state inactivation was more negative and recovery from inactivation was delayed. Co-expression of DPPX in addition to Kv4.3 and KChIP2a produced similar current kinetics as in human ventricular myocytes. We therefore propose that DPPX is an essential component of the native cardiac I(to) channel complex in human heart. PMID- 15890704 TI - NaPi-IIa and interacting partners. AB - Regulation of renal proximal tubular reabsorption of phosphate (Pi) is one of the critical steps in Pi homeostasis. Experimental evidence suggests that this regulation is achieved mainly by controlling the apical expression of the Na+ dependent Pi cotransporter type IIa (NaPi-IIa) in proximal tubules. Only recently have we started to obtain information regarding the molecular mechanisms that control the apical expression of NaPi-IIa. The first critical observation was the finding that truncation of only its last three amino acid residues has a strong effect on apical expression. A second major finding was the observation that the last intracellular loop of NaPi-IIa contains sequence information that confers parathyroid hormone (PTH) sensitivity. The use of the above domains of the cotransporter in yeast two-hybrid (Y2H) screening allowed the identification of proteins interacting with NaPi-IIa. Biochemical and morphological, as well as functional, analyses have allowed us to obtain insights into the physiological roles of such interactions, although our present knowledge is still far from complete. PMID- 15890705 TI - Kv7/KCNQ/M and HCN/h, but not KCa2/SK channels, contribute to the somatic medium after-hyperpolarization and excitability control in CA1 hippocampal pyramidal cells. AB - In hippocampal pyramidal cells, a single action potential (AP) or a burst of APs is followed by a medium afterhyperpolarization (mAHP, lasting approximately 0.1 s). The currents underlying the mAHP are considered to regulate excitability and cause early spike frequency adaptation, thus dampening the response to sustained excitatory input relative to responses to abrupt excitation. The mAHP was originally suggested to be primarily caused by M-channels (at depolarized potentials) and h-channels (at more negative potentials), but not SK channels. In recent reports, however, the mAHP was suggested to be generated mainly by SK channels or only by h-channels. We have now re-examined the mechanisms underlying the mAHP and early spike frequency adaptation in CA1 pyramidal cells by using sharp electrode and whole-cell recording in rat hippocampal slices. The specific M-channel blocker XE991 (10 microm) suppressed the mAHP following 1-5 APs evoked by current injection at -60 mV. XE991 also enhanced the excitability of the cell, i.e. increased the number of APs evoked by a constant depolarizing current pulse, reduced their rate of adaptation, enhanced the after depolarization and promoted bursting. Conversely, the M-channel opener retigabine reduced excitability. The h channel blocker ZD7288 (4-ethylphenylamino-1,2-dimethyl-6-methylaminopyrimidinium chloride; 10 microm) fully suppressed the mAHP at -80 mV, but had little effect at -60 mV, whereas XE991 did not measurably affect the mAHP at -80 mV. Likewise, ZD7288 had little or no effect on excitability or adaptation during current pulses injected from -60 mV, but changed the initial discharge during depolarizing pulses injected from -80 mV. In contrast to previous reports, we found that blockade of Ca2+-activated K+ channels of the SK/KCa type by apamin (100-400 nm) failed to affect the mAHP or adaptation. A computational model of a CA1 pyramidal cell predicted that M- and h-channels will generate mAHPs in a voltage-dependent manner, as indicated by the experiments. We conclude that M- and h-channels generate the somatic mAHP in hippocampal pyramidal cells, with little or no net contribution from SK channels. PMID- 15890706 TI - Inspiratory phase-locked alpha oscillation in human olfaction: source generators estimated by a dipole tracing method. AB - Olfactory perception and related emotions are largely dependent on inspiration. We acquired simultaneous respiration and electroencephalographic recordings during pleasant odour and unpleasant odour stimulation. We sought to identify changes in respiratory pattern, inspiratory-related potentials and location of dipoles estimated from the potentials. Electroencephalographic recording was triggered by inspiration onset. Respiratory frequency decreased at pleasant odour recognition, and it increased at unpleasant odour detection and recognition. O2 consumption records showed that these changes were not due to metabolic demand. During olfactory stimulation, inspiratory phase-locked alpha oscillation (I alpha) was found in the averaged potential triggered by inspiration onset. I alpha was observed at both pleasant odour and unpleasant odour detection and recognition, but it was not seen in the inspiration-triggered potentials of normal air breathing. Electroencephalographic dipole tracing identified the location of dipoles from the I-alpha in the limbic area and the cortex; the entorhinal cortex, hippocampus, amygdala, premotor area and centroposterior orbitofrontal cortex subserve odour detection, and the rostromedial orbitofrontal cortex subserves odour recognition. We suggest that the I-alpha in our study originated from the olfactory cortex in the forebrain and was phase-locked to inspiration. PMID- 15890707 TI - Down regulation of Kv3.4 channels by chronic hypoxia increases acute oxygen sensitivity in rabbit carotid body. AB - The carotid body (CB) chemoreceptors participate in the ventilatory responses to acute and chronic hypoxia (CH). Arterial hypoxaemia increases breathing within seconds, and CB chemoreceptors are the principal contributors to this reflex hyperventilatory response. Acute hypoxia induces depolarization of CB chemoreceptors by inhibiting certain K+ channels, but the role of these channels in CH, as in high-altitude acclimatization, is less known. Here we explored the effects of prolonged (24-48 h) hypoxic exposure of rabbit CB chemoreceptor cells in primary cultures on the voltage-dependent K+ currents and on their response to acute hypoxia. We found that CH induces a decrease in the amplitude of outward K+ currents due to a reduction in a fast-inactivating BDS- and highly TEA-sensitive component of the current. In spite of this effect, acute hypoxic inhibition of K+ currents is increased in CH cultures, as well as hypoxia-induced depolarization. These data suggest that downregulation of this component (that does not contribute to the oxygen-sensitive K+ current (IKO2) participates in the hypoxic sensitization. Pharmacological, immunocytochemical and quantitative PCR (qPCR) experiments demonstrate that CH-induced decrease in outward K+ currents is due to a downregulation of the expression of Kv3.4 channels. Taken together, our results suggest that CH sensitization in rabbit CB could be achieved by an increase in the relative contribution of IKO2 to the outward K+ current as a consequence of the decreased expression of the oxygen-insensitive component of the current. We conclude that acute and chronic hypoxia can exert their effects acting on different molecular targets. PMID- 15890709 TI - No genetic association of the human prolyl endopeptidase gene in the Dutch celiac disease population. AB - Celiac disease (CD) is a complex genetic disorder of the small intestine. The DQ2/DQ8 human leucocyte antigen (HLA) genes explain approximately 40% of the genetic component of the disease, but the remaining non-HLA genes have not yet been identified. The key environmental factor known to be involved in the disease is gluten, a major protein present in wheat, barley, and rye. Integrating microarray data and linkage data from chromosome 6q21-22 revealed the prolyl endopeptidase (PREP) gene as a potential CD candidate in the Dutch population. Interestingly, this gene encodes for the only enzyme that is able to cleave the proline-rich gluten peptides. To investigate the role of the human PREP gene as a primary genetic factor in CD, we conducted gene expression, sequence analysis, and genetic association studies of the PREP gene and determined PREP enzyme activity in biopsies from CD patients and controls. Sequence analysis of the coding region of the PREP gene revealed two novel polymorphisms. Genetic association studies using two novel polymorphisms and three known PREP variants excluded a genetic association between PREP and CD. Determination of PREP activity revealed weak but significant differences between treated and untreated CD biopsies (P < 0.05). Our results from the association study indicate that PREP is not a causative gene for CD in the Dutch population. These are further supported by the activity determinations in which we observed no differences in PREP activity between CD patients and controls. PMID- 15890708 TI - Autonomic nervous system influence on arterial baroreflex control of heart rate during exercise in humans. AB - A combination of sympathoexcitation and vagal withdrawal increases heart rate (HR) during exercise, however, their specific contribution to arterial baroreflex sensitivity remains unclear. Eight subjects performed 25 min bouts of exercise at a HR of 90, 120, and 150 beats min-1, respectively, with and without metoprolol (0.16 +/- 0.01 mg kg(-1); mean +/- S.E.M.) or glycopyrrolate (12.6 +/- 1.6 microg kg-1). Carotid baroreflex (CBR) function was determined using 5 s pulses of neck pressure (NP) and neck suction (NS) from +40 to -80 Torr, while transfer function gain (GTF) was calculated to assess the linear dynamic relationship between mean arterial pressure and HR. Spontaneous baroreflex sensitivity (SBR) was evaluated as the slope of sequences of three consecutive beats in which systolic blood pressure and the R-R interval of the ECG either increased or decreased, in a linear fashion. The beta-1 adrenergic blockade decreased and vagal cardiac blockade increased HR both at rest and during exercise (P < 0.05). The gain at the operating point of the modelled reflex function curve (GOP) obtained using NP and NS decreased with workload independent of beta-1 adrenergic blockade. In contrast, vagal blockade decreased GOP from -0.40 +/- 0.04 to -0.06 +/- 0.01 beats min-1 mmHg-1 at rest (P < 0.05). Furthermore, as workload increased both GOP and SBR, and GOP and GTF were correlated (P < 0.001), suggesting that the two dynamic methods applied to evaluate arterial baroreflex (ABR) function provide the same information as the modelled GOP. These findings suggest that during exercise the reduction of arterial baroreceptor reflex sensitivity at the operating point was a result of vagal withdrawal rather than an increase in sympathetic activity. PMID- 15890710 TI - Carbon monoxide protects against the development of experimental necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is a disease of neonates that is increasing in incidence and often results in significant morbidity and mortality. Carbon monoxide (CO), a byproduct of the catabolism of heme, is known to have anti inflammatory and antiapoptotic properties. In this study, we aimed to demonstrate that inhaled CO protects against the development of intestinal inflammation in a model of experimental NEC as well as decreases enterocyte cell death in vitro. Additionally, we also aimed to demonstrate that CO decreases enterocyte production of inducible nitric oxide synthase (iNOS) and nitric oxide (NO). Neonatal rats were exposed to intermittent hypoxia exposure and formula feeding to induce experimental NEC. Animals randomized to CO treatment were put in an environment containing 0.025% CO for 1 h/day on days 1-3 of life. All animals were killed on day 4 of life. In vitro experiments were performed with IEC-6 cells, a rat enterocyte cell line. Cells were examined for viability, iNOS production, and elaboration of NO. We found that CO diminished levels of serum inflammatory cytokines and nitrites, protected against intestinal inflammation, and decreased ileal iNOS production and protein nitration in a model of experimental NEC. In vitro, CO decreased cytokine- or hypoxia/endotoxin-induced iNOS and NO production. CO also abrogated TNF-alpha- and actinomycin D-induced apoptosis or hypoxia/endotoxin-induced cell death. In conclusion, 1 h of daily low-dose inhaled CO protected against the development of intestinal inflammation in a model of experimental NEC. iNOS and NO production were decreased by CO both in vivo and in vitro. CO may prove to be a useful clinical adjunct in the treatment of NEC. PMID- 15890711 TI - Platelet-activating factor and prostaglandin E2 impair esophageal ACh release in experimental esophagitis. AB - ACh is a neurotransmitter in cat esophageal circular muscle, as atropine nearly abolishes contraction of in vitro circular muscle strips in response to electric field stimulation (EFS) (5, 12). Experimental esophagitis reduced EFS- but not ACh-induced contraction of esophageal circular muscle, suggesting that esophagitis impairs neurotransmitter release. Because IL-1beta and IL-6 are produced in esophagitis and reproduce these changes in normal esophageal muscle (12), we examined the role of IL-1beta and IL-6 in this motor dysfunction. IL 1beta, IL-6 (12), H2O2, PGE2, and platelet-activating factor (PAF) were elevated in esophagitis specimens. Normal muscle incubated (2 h) in IL-1beta and IL-6 had increases in H2O2, PGE2, and PAF levels. H2O2 contributed to increased PGE2 and PAF, as the increase was partially (60-80%) reversed by the H2O2 scavenger catalase. EFS-induced [3H]ACh release from muscle strips significantly (42%) decreased in esophagitis and after 2 h incubation in PGE2 and in PAF C-16. Similarly, EFS-induced but not ACh-induced muscle contraction decreased in esophagitis and after incubation in PGE2 and PAF C-16. Finally, in normal muscle strips treated with IL-1beta electrical field stimulation (EFS)-induced contraction was partially restored by indomethacin or by the PAF antagonist CV3988 and was completely restored by the combination of CV3988 and indomethacin, whereas in strips treated with IL-6, EFS-induced contraction was partially restored by the PAF antagonist CV3988 and not affected by indomethacin. We conclude that IL-1beta-induced production of H2O2 causes formation of PGE2 and PAF that inhibit ACh release from esophageal cholinergic neurons without affecting ACh-induced contraction of esophageal circular muscle. IL-6 causes production of H2O2, PAF, and other unidentified inflammatory mediators. PMID- 15890712 TI - Events at the host-microbial interface of the gastrointestinal tract III. Cell-to cell signaling among microbial flora, host, and pathogens: there is a whole lot of talking going on. AB - Humans have an important association with their intestinal microbial flora. The microbial flora helps to shape the mammalian innate immune system, absorbs nutrients, and plays an intricate role on intestinal development. Microbes and mammals communicate with each other through an array of hormone and hormonelike chemical compounds. These "signals," however, are hijacked by bacterial pathogens, such as enterohemorrhagic Eschrichia coli (EHEC), to activate its virulence genes, colonize the host, and start the disease process. This review explores the cell-to-cell signaling events in the gastrointestinal tract that lead EHEC to regulate its virulence genes in a coordinate manner. PMID- 15890713 TI - Transient upregulation of GRP and its receptor critically regulate colon cancer cell motility during remodeling. AB - Gastrin-releasing peptide (GRP) is typically viewed as a growth factor in cancer. However, we have suggested that in colon cancer, GRP acts primarily as a morphogen when it and its receptor (GRP-R) are aberrantly upregulated. As such, GRP/GRP-R act(s) primarily to modulate processes contributing to the assumption or maintenance of tumor differentiation. One of the most important such processes is the ability of tumor cells to achieve directed motility in the context of tissue remodeling. Yet the cellular conditions affecting GRP/GRP-R expression, and the biochemical pathways involved in mediating its morphogenic properties, remain to be established. To study this, we evaluated the human colon cancer cell lines Caco-2 and HT-29 cells. We found that confluent cells do not express GRP/GRP-R. In contrast, disaggreation and plating at subconfluent densities results in rapid GRP/GRP-R upregulation followed by their progressive decrease as confluence is achieved. GRP/GRP-R coexpression correlated with that of focal adhesion kinase (FAK) phosphorylation of Tyr(397), Tyr(407), Tyr(861), and Tyr(925) but not Tyr(576) or Tyr(577). To more specifically evaluate the kinetics of GRP/GRP-R upregulation, we wounded confluent cell monolayers. At t = 0 h GRP/GRP-R were not expressed, yet cells immediately began migrating into the gap created by the wound. GRP/GRP-R were first detected at approximately 2 h, and maximal levels were observed at approximately 6 h postwounding. The GRP-specific antagonist [d-Phe(6)]-labeled bombesin methyl ester had no effect on cell motility before GRP-R expression. In contrast, this agent increasingly attenuated cell motility with increasing GRP-R expression such that from t = 6 h onward no further cell migration into the gap was observed. Overall, these findings indicate the existence of GRP-independent and -dependent phases of tumor cell remodeling with the latter mediating colon cancer cell motility during remodeling via FAK. PMID- 15890714 TI - Binding of acellular, native and cross-linked human hemoglobins to haptoglobin: enhanced distribution and clearance in the rat. AB - It is well established that hemoglobin resulting from red cell lysis binds to haptoglobin in plasma to form a complex. The increased molecular size precludes its filtration by the kidneys, redirecting it toward hepatocellular entry. Chemically cross-linked hemoglobins are designed to be resistant to renal excretion, even in the absence of haptoglobin. The manner in which binding to haptoglobin influences the pharmacokinetics of acellular cross-linked and native hemoglobins was investigated after intravenous injection of radiolabeled native human hemoglobin and trimesyl-(Lys82)beta-(Lys82)beta cross-linked human hemoglobin, at trace doses, into rats. Under these conditions, there is sufficient plasma haptoglobin for binding with hemoglobin. In vitro binding assayed by size-exclusion chromatography for bound and free hemoglobin revealed that, at <8 muM hemoglobin, native human hemoglobin was completely bound to rat haptoglobin, whereas only approximately 30% of trimesyl-(Lys82)beta-(Lys82)beta cross-linked hemoglobin was bound. Plasma disappearance of low doses (0.31 mumol/kg) of native and cross-linked hemoglobins was monoexponential (half-life = 23 and 33 min, respectively). The volume of distribution (40 vs. 19 ml/kg) and plasma clearance (1.22 vs. 0.4 ml.min(-1).kg(-1)) were higher for native than for cross-linked hemoglobin. Native and cross-linked human hemoglobins were found primarily in the liver, and not in the kidney, heart, lung, or spleen, mostly as degradation products. These pharmacokinetic findings suggest that the binding of hemoglobin to haptoglobin enhances its hepatocellular entry, clearance, and distribution. PMID- 15890715 TI - Prevalence of Ambler class A and D beta-lactamases among clinical isolates of Pseudomonas aeruginosa in Korea. AB - OBJECTIVES: Recently, resistance to extended-spectrum cephalosporins due to acquired beta-lactamases has been reported in Pseudomonas aeruginosa. The aim of this study was to investigate the prevalence of Ambler class A and D beta lactamases and their extended-spectrum derivatives and antimicrobial susceptibilities of P. aeruginosa isolated from various parts of Korea. METHODS: A total of 252 consecutive, non-duplicate isolates of P. aeruginosa were studied for the presence of class A or D beta-lactamase. Antibiotic susceptibility tests and PCR amplification of genes encoding class A (bla(PSE-1), bla(PER-1), bla(VEB 1), bla(TEM), bla(SHV), bla(CTX-M) and bla(GES-1)) and class D beta-lactamases (bla(OXA-groupI), bla(OXA-groupII) and bla(OXA-groupIII)) were performed. For PCR positive isolates, isoelectric focusing (IEF) analysis, sequencing and pulsed field gel electrophoresis (PFGE) were performed. RESULTS: In 64 (25.4%) isolates, structural genes for PSE-1 (6.3%), OXA-10 (13.1%), OXA-4 (4.3%), OXA-30 (2.0%), OXA-2 (2.3%) and OXA-17 (0.4%) were found; their distribution varied between provinces. None harboured bla(PER-1), bla(VEB-1), bla(TEM), bla(SHV), bla(CTX-M) and bla(GES-1). The cross-class resistance rates to other antibiotics was significantly higher in class A and D beta-lactamase producers than in non producers (P < 0.001 for aminoglycosides, ciprofloxacin and meropenem). CONCLUSIONS: OXA-type beta-lactamases are widespread, but their extended-spectrum derivatives are rare among P. aeruginosa in Korea. To our knowledge, this is the first report of OXA-17, an extended-spectrum derivative of OXA-10, outside the Middle East. In addition, combined resistance to ticarcillin and aminoglycosides was a useful indicator for P. aeruginosa producing PSE- or OXA-type beta lactamases in this study. PMID- 15890716 TI - Risk factors for antibiotic-resistant Escherichia coli isolated from community acquired urinary tract infections in Dakar, Senegal. AB - OBJECTIVES: To assess overall resistance rates and risk factors for resistance to ampicillin, co-amoxiclav, nalidixic acid, fluoroquinolones and trimethoprim/sulfamethoxazole in Escherichia coli strains isolated from outpatients with acute urinary tract infection in Dakar (Senegal). PATIENTS AND METHODS: From June 2001 to June 2003, a prospective study was performed among Senegalese outpatients consulting at the Institut Pasteur of Dakar for urine analysis. Evaluated risk factors were: age, gender, prior hospitalization, antibiotic exposure, urinary tract infection and urinary catheter. RESULTS: A total of 398 non-duplicate, consecutive, biologically significant E. coli were isolated. The levels of antibiotic resistance in Dakar appeared dramatic and worrisome with resistance rates ranging from 18.6% for fluoroquinolones to 73.6% for ampicillin. With the exception of the presence of urinary catheter, the risk factors identified were consistent with data previously reported in developed countries. CONCLUSIONS: We hope our results will assist medical authorities in the development of appropriate control strategies. PMID- 15890717 TI - Characterization of a multidrug-resistant isolate of Salmonella Paratyphi B from Japan. PMID- 15890718 TI - Impact of carbon dioxide on the susceptibility of key respiratory tract pathogens to telithromycin and azithromycin. AB - OBJECTIVES: To determine the quantitative differences in telithromycin and azithromycin MIC values against Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes obtained using two recommended and commonly used methodologies: CLSI reference standard broth microdilution in ambient air and Etest((R)) concentration gradient in CO(2). METHODS: Four hundred clinical isolates (S. pneumoniae, n = 200; H. influenzae, n = 100; S. pyogenes, n = 100) were evaluated in seven independent laboratories. Telithromycin and azithromycin MICs were determined using CLSI broth microdilution panels incubated in ambient air and Etest strips incubated in CO(2). Standard quality control reference strains-S. pneumoniae ATCC 49619 (n = 10) and H. influenzae ATCC 49247 (n = 10) were also tested. RESULTS: Telithromycin and azithromycin Etest MICs in CO(2) were elevated for all organisms when compared with values obtained using broth microdilution in ambient air. Telithromycin geometric mean MIC values increased in CO(2) by 2.05, 1.00 and 1.78 log(2) dilutions for S. pneumoniae, H. influenzae and S. pyogenes, respectively. The corresponding values for azithromycin were 2.54, 1.21 and 3.0 log(2) dilutions, respectively. CONCLUSIONS: Telithromycin MICs measured using Etest in CO(2) are consistently elevated compared with those generated by CLSI broth microdilution measured in ambient air. These findings indicate that Etest should not be routinely used for the determination of telithromycin MICs against S. pneumoniae, H. influenzae and S. pyogenes, unless appropriate corrective factors are applied before reporting MICs or applying interpretive susceptibilities. Based on results from this study, Etest MIC breakpoints and quality control ranges are proposed. PMID- 15890719 TI - Audit of antibiotic policies in the South East of England, 2004. AB - OBJECTIVES: The antibiotic policies of hospitals and primary care trusts (PCTs) in South East England were audited in the summer of 2004, to see how they had improved since 2000. METHODS: Antibiotic policies were obtained from pharmacists in NHS hospitals and PCTs, and examined for dates, formats, evidence base for policies, the type of guidance given on dosage, length of treatment, choice of antibiotics, coverage of common infections and reasons for prophylaxis. RESULTS: Twenty-three hospital and 25 primary care policies were examined. The average age of policies was 12 months, but 13 were more than 2 years old. The commonest format was an A4-sized document available in an electronic version. Primary care policies were more uniform than hospital policies. More primary care than hospitals' policies gave evidence to support their guidance. Ten policies used plain English for dosages, and 38 (79%) policies made few or no cautionary points about the drugs recommended. Respiratory and urinary infections were covered in most policies, but guidance on gastroenteritis and antibiotic prophylaxis was less frequent. There was little advice in the policies on the management of methicillin-resistant Staphylococcus aureus. CONCLUSIONS: Primary care policies have improved since 2000, using a national model for evidence and a consistent style. Hospitals could benefit from similar national guidance, especially in the evidence to support the contents of antibiotic policies. PMID- 15890720 TI - Hyperproduction of AmpC beta-lactamase in a clinical isolate of Escherichia coli associated with a 30 bp deletion in the attenuator region of ampC. PMID- 15890721 TI - Influence of inoculum size of Staphylococcus aureus and Pseudomonas aeruginosa on in vitro activities and in vivo efficacy of fluoroquinolones and carbapenems. AB - OBJECTIVES: We investigated the effect of inoculum size on MIC, bactericidal activity and the post-antibiotic effect (PAE) of carbapenems (imipenem, panipenem and meropenem) and injectable quinolones (pazufloxacin and ciprofloxacin) against Staphylococcus aureus and Pseudomonas aeruginosa, and also the relationship between in vivo systemic infection by changing the inoculum size injected in mice. RESULTS: Increasing the bacterial inoculum (10(5)-10(8) cfu/mL) had no significant effect on the MIC of any of the tested antimicrobial agents. With the standard inocula (10(6) cfu/mL) of both test strains, all the antimicrobial agents showed bactericidal activity; however, increasing the inoculum size to >10(8) cfu/mL resulted in a reduction in bactericidal activity of all the antimicrobial agents against S. aureus Smith. In contrast, increasing the inoculum size of P. aeruginosa exerted only a minimal influence on the bactericidal activity of fluoroquinolones, but resulted in a reduction in the bactericidal activity of carbapenems. With the standard inoculum size of S. aureus Smith, pre-incubation with fluoroquinolones and carbapenems, except for meropenem, was sufficient to produce PAEs. When the inoculum was increased, the duration of the PAEs of these antimicrobial agents was reduced; however, those of fluoroquinolones were longer than carbapenems. Inoculum size had a greater influence on the in vivo efficacy of carbapenems than that of fluoroquinolones. CONCLUSIONS: Our results suggest that decreased bactericidal activity, or the in vitro PAE of carbapenems and fluoroquinolones, is related to the reduced in vivo protective effect against infection caused by high inoculum with S. aureus or P. aeruginosa. PMID- 15890722 TI - Bronchiectasis: the consequence of late diagnosis in chronic respiratory symptoms. AB - Bronchiectasis is still common among some developing countries like Turkey. The aim of this study was to document the number of children with non-cystic fibrosis (CF) bronchiectasis, to evaluate the risk factors and to emphasize early diagnosis and treatment. All children, except those diagnosed with CF, with bronchiectasis established by chest radiogram, bronchography and/or computed tomography or biopsy material, were retrospectively reviewed. They were tested for serum total eosinophil count, nasal smear, serum levels of immunoglobulins A, G, M, E, and serum alpha-1 antitrypsin level. Pulmonary function tests, rigid bronchoscopy, nasal biopsy, lung scintigraphy, and echocardiogram were also performed. There were 204 patients whose most common presenting symptoms were cough, sputum expectoration, and dyspnea. Bronchiectasis was present mostly in the left lower lobe. The cause could not be determined in 49 per cent of patients. Among the identified causes, infection was present in most patients, followed by asthma, primary ciliary dyskinesia, congenital immune deficiency, and foreign body aspiration. It is possible to prevent bronchiectasis in children with vaccinations and improved nutrition in developing countries. Early diagnosis and treatment will increase the quality of life and survival of patients with bronchiectasis, which has irreversible and progressive complications if untreated. PMID- 15890723 TI - Validation of a new clinical case definition for paediatric HIV infection, Bloemfontein, South Africa. PMID- 15890724 TI - Antibiotic use in community-based pediatric outpatients in southern region of Brazil. AB - The irrational use of antibiotics is incriminated for the escalating antimicrobial resistance problem worldwide. This cross-sectional survey was conducted to describe patterns of antibiotic use in community-based pediatric outpatients in the southern region of Brazil. A sample of children (29 days to 18 years of age) was selected from each community-based outpatient clinic. Information related to antibiotic use in the child was obtained by interview with their parents using a structured questionnaire. Of 489 children, 201 (41.1 per cent) had received at least one antimicrobial course in the previous 2 months. The most commonly prescribed antibiotic was amoxicillin (54.0 per cent) and broad spectrum agents were used in 15.3 per cent of antimirobial courses. Acute respiratory infections were the most frequent diagnosis for antibiotic prescribing (91.2 per cent), but at least 39.5 per cent were conditions of presumed viral etiology. Antibiotic use in the previous 2 months was more likely in children younger than 2 years (PR 1.36, 95 per cent CI 1.10-1.69); with chronic diseases (PR 1.38, 95 per cent CI 1.10-1.75); with higher paternal education level (PR 1.38, 95 per cent CI 1.11-1.72); with parents' pressure on physician for antibiotics (PR 1.66, 95 per cent CI 1.05-2.64) and with parental self-prescribing habit (PR 1.47, 95 per cent CI 1.06-2.04). Antibiotic use in children increased with mother's age (p=0.03 for linear trend). In conclusion, antibiotic prescribing is very common in community-based pediatric outpatients in the southern region of Brazil and inappropriate use is frequent. The independent risk factors for antibiotic use are child's age under 2 years, the presence of chronic diseases in the child, older mother, higher paternal education level, parents' pressure on physician and parental self-prescribing habit. PMID- 15890725 TI - Attitudes towards exclusive breastfeeding and other infant feeding options--a study from Abidjan, Cote d'Ivoire. AB - The HIV/AIDS-pandemic causes many problems for the most affected societies and their health care systems. One of these is the 'parent to child transmission' (PTCT) through breastmilk and its prevention (PPTCT). As economic and hygienic conditions do not always assure safe replacement feeding in developing countries, a WHO/UNAIDS/UNICEF-expert panel proposed methods to reduce the risk of PTCT but to use breastmilk for infant feeding. The study presented here aimed at identifying the expected acceptance of such a concept by addressing the attitudes of women in Abidjan, Ivory Coast. Interviews were performed with 150 mothers and 60 pregnant women. The vast majority regarded breastfeeding as the appropriate method of infant feeding, although the idea of exclusive breastfeeding was not well accepted. Water, especially, was felt to be a necessary supplement. In case of a suggested HIV-infection of the mother, 74 per cent of the women voted for weaning after 3 months. Eighty-three per cent accepted the exclusive use of breastmilk substitutes from birth. Seventy-six per cent were ready to boil their milk for pasteurization. Only 37 per cent considered a wet-nurse to breastfeed their child. As mixed breastfeeding implies a highest risk of PTCT of HIV, the most favoured option-exclusive breastfeeding and early weaning-requires some effort to convince women that breastmilk is a sufficient source of nutrients, fluid and energy for their child and that this feeding should preferably be practised up to 6 months of age. For affluent women, breastmilk substitutes can also be considered as a means of PPTCT in overall resource-poor countries. For the majority of women, there is no real alternative to breastfeeding and to the use of breastmilk for which appropriate technologies of PPTCT are to be developed with respect to national, local and household specifications. PMID- 15890726 TI - Gangrene in a child with Plasmodium falciparum malaria. PMID- 15890727 TI - Usefulness and limits of CA-125 in diagnosis of endometriosis without associated ovarian endometriomas. AB - BACKGROUND: The aim of this study was to evaluate the diagnostic significance of CA-125 for endometriosis without ovarian endometriomas. METHODS: Preoperative serum CA-125 levels were measured in 775 consecutive women diagnosed by laparoscopy or laparotomy with endometriosis, adenomyosis, leiomyomas, or normal pelvis. RESULTS: Receiver operating characteristic curve analysis revealed that the area under the curve for endometriosis without endometriomas was 0.788, significantly smaller than that for endometriosis with endometriomas (0.935, P < 0.05). In diagnosis of endometriosis without endometriomas, both the maximal accuracy of 78.8% and the maximal diagnostic value of 61.2% were obtained at the cutoff value of 20 U/mL. Negative predictive value was 78.0% at the cutoff value of 20 U/mL, whereas positive predictive value was 92.9% at the cutoff value of 30 U/mL. This range is clearly superior to the empirical single cutoff of 35 U/mL. CONCLUSIONS: In the diagnosis of endometriosis without endometriomas, combined use of two cutoff values for CA-125, 20 and 30 U/mL, provides improved diagnostic performance. However, the accuracy of using only CA-125 testing for diagnosis is still limited. Serum CA-125 testing can be done during initial screenings of women with possible endometriosis. PMID- 15890728 TI - 'Vanishing embryo syndrome' in IVF/ICSI. AB - BACKGROUND: In a Danish population-based cohort study assessing the risk of cerebral palsy in children born after IVF, we made some interesting observations regarding 'vanishing co-embryos'. METHODS AND RESULTS: All live-born children born in Denmark from 1 January 1995 to 31 December 2000 were included in this analysis. The children conceived by IVF/ICSI (9444) were identified through the IVF Register, the children conceived without IVF/ICSI (395 025) were identified through The Danish Medical Birth Register. Main outcome measure was the incidence of cerebral palsy. Within the IVF/ICSI children we found indications of an increased risk of cerebral palsy in those children resulting from pregnancies, where the number of embryos transferred was higher than the number of children born. CONCLUSIONS: The association between vanishing embryo syndrome and incidence of cerebral palsy following IVF requires further investigation in larger, adequately powered, studies. PMID- 15890729 TI - Does the potential for selection bias in semen quality studies depend on study design? Experience from a study conducted within an infertility clinic. AB - BACKGROUND: The low participation rates in human semen quality studies raises concern for the potential of differential participation based on semen quality (or a surrogate). To explore the potential for differential participation, we compared semen analysis results from study subjects with those of non-study subjects. METHODS: We obtained semen analysis results from 235 study subjects and retrospectively obtained results from a subset of 235 infertility clinic patients that were not study subjects but met the same eligibility criteria. The study was conducted at the Massachusetts General Hospital Infertility Clinic. All semen samples (study subjects and non-study subjects) were analysed for sperm concentration and motility by computer-aided semen analysis (CASA), and morphology was assessed using strict criteria. Semen analysis parameters for the non-study subjects were compared with the semen analysis results from study subjects. RESULTS: For all semen characteristics (sperm concentration, total sperm count, sperm motility and morphology), there were only marginal (non significant) differences between study subjects and non-study subjects. CONCLUSIONS: Among men from an infertility clinic, we found no strong evidence of differential participation based on semen quality. This is reassuring since the potential for selection bias is of concern in semen quality studies. However, the potential for selection bias in other study designs remains unclear. PMID- 15890730 TI - Hysteroscopy may be justified after two miscarriages. AB - BACKGROUND: Recurrent pregnancy loss is traditionally investigated after three or more consecutive pregnancy losses. Although there is a trend to start investigation after two miscarriages, data are not available to date to justify this approach. We sought to compare the frequency of uterine anomalies between women referred to hysteroscopy for repeated miscarriages after two, and three or more, miscarriages respectively. METHODS: A retrospective analysis of acquired and congenital uterine anomalies in all patients undergoing hysteroscopy for repeated pregnancy loss at an academic and referral medical centre. RESULTS: Hysteroscopy was performed on 165 women referred for recurrent pregnancy loss: 67 after two and 98 after three or more consecutive miscarriages. The rate of uterine anomalies did not differ significantly and was 32 versus 28% respectively. CONCLUSIONS: Hysteroscopy may be justified following two spontaneous pregnancy losses. PMID- 15890731 TI - Comparison of GnRH agonists and antagonists in assisted reproduction cycles of patients at high risk of ovarian hyperstimulation syndrome. AB - BACKGROUND: During IVF or ICSI cycles, ovarian hyperstimulation syndrome (OHSS) is a major problem. The aim of this prospective, multicentre, comparative study (using historical controls) was to assess the efficacy of a GnRH antagonist protocol in preventing OHSS in selected patients who had experienced OHSS or had been at risk of OHSS in their previous IVF/ICSI attempt. METHODS AND RESULTS: Patients underwent a new cycle where the same gonadotrophin protocol was used [same dose of recombinant FSH (rFSH)] but a different protocol was used for pituitary desensitization: cetrorelix 0.25 mg multiple-dose antagonist instead of GnRH agonist long protocol. Cetrorelix 0.25 mg was administered daily, starting when the leading follicle reached a diameter of 14 mm. In other words, rFSH was administered in the new cycle according to the dosage and the step-up or step down modalities used during the previous cycle, independently of ultrasound findings and serum estradiol (E(2)) levels. Eighty-seven patients entered the study. Out of the 87 cycles involving GnRH agonists, 49 (56.3%) were cancelled and out of the 87 involving GnRH antagonists 28 (32.2%) were cancelled [McNemar's test; 95% confidence interval (CI) -35.8% to -11.2%; P < 0.001]. After GnRH agonist cycles, we recorded 24 cases of OHSS (18 moderate and six severe; 27.6%), whereas after the GnRH antagonist cycles there were 10 cases of OHSS (nine moderate and one severe; 11.5%) (95% CI-26.4% to -5.7%; P = 0.006). There was a statistically significant reduction in the total number of follicles with a diameter >10 mm (Wilcoxon's test; Z = 6.1; P < 0.001) and of E(2) levels on the day of HCG administration (2538 versus 4322.4 pg/ml; P < 0.001) in the GnRH antagonist cycles versus GnRH agonist cycles. Twenty-nine patients had an embryo transfer in the first cycle (76.3% of oocyte retrievals) and 57 in the cycle using GnRH antagonist (96.6%). This 20.3% difference was also significant (Z test; 95% CI 6.8-36.0%; P = 0.003). After the antagonist cycles, 18 pregnancies (20.7 per initiated cycle; 31.6% per embryo transfer) were obtained. CONCLUSIONS: Although this study presents some limitations owing to the use of historical controls, our data show a favourable effect of GnRH antagonists in reducing the incidence of OHSS and the number of assisted fertilization cycles cancelled because of the risk of OHSS in high responder patients. As a consequence, GnRH antagonist plus gonadotrophin administration could also increase the percentage of oocyte retrievals and embryo transfers in this high risk group of patients. PMID- 15890732 TI - Reduced semen quality in chronic prostatitis patients that have cellular autoimmune response to prostate antigens. AB - BACKGROUND: The relationship between chronic prostatitis and fertility has been controversial for many years. We have previously shown the presence of a cellular autoimmune response against prostate antigens in a group of chronic prostatitis patients. Our main goal was to investigate whether chronic prostatitis (either caused by an infection or an autoimmune response to the prostate gland) could have a deleterious effect on semen quality. METHODS: Forty-four patients diagnosed as suffering from chronic prostatitis were included and divided into groups according to the presence of infection and/or cellular autoimmune response against prostate antigens. Healthy normal individuals were included as controls. Measurements for sperm concentration, motility, morphology, prostate and seminal vesicle markers, antisperm antibodies, white blood cells and pro-inflammatory cytokines were performed accordingly. RESULTS: The most severe abnormalities were seen in patients with no evident infection and an autoimmune response against prostate antigens. Moreover, significantly increased levels of pro-inflammatory cytokines were detected in seminal plasma from these patients. CONCLUSIONS: This study shows that chronic prostatitis patients with cellular autoimmune response to prostate antigens present important alterations in their semen quality parameters. We speculate that an autoimmune response against prostate antigens and the inflammatory process involved may affect male fertility. PMID- 15890733 TI - Is granulocyte colony-stimulating factor level predictive for human IVF outcome? AB - BACKGROUND: We investigated granulocyte colony-stimulating factor (G-CSF) in human reproduction. METHODS: From a total sample of 93 patients, we analysed in group 1 (n = 82) the level of G-CSF and estradiol (E(2)) in serum and follicular fluid (FF) on day of follicular puncture (FP). Furthermore, in response to ovarian stimulation, G-CSF levels in serum were compared between low (n = 11), moderate (n = 53) and high (n = 18) response patients. In group 2 (n = 23) serum for G-CSF assessment was collected throughout menstrual cycle until gestation. Group 3 (n = 11) patients with endometriosis were assessed for G-CSF in serum and FF on day of FP without further differentiation. RESULTS: G-CSF in FF was higher than in serum (P < 0.01). G-CSF in serum increased from low through moderate to high response (P < 0.001); pregnancy rates were 0, 24.5 and 33.5% respectively. G CSF in serum increased throughout stimulation, reached a peak with ovulation induction (P = 0.01) and decreased until embryo transfer (P=0.001). G-CSF level only in pregnant patients (n = 11) increased from embryo transfer to implantation to gestation (P = 0.005). In endometriosis patients G-CSF in serum and FF was lower than in non-endometriosis patients (P < or = 0.03) and corresponded with low response patients. CONCLUSIONS: G-CSF is involved in follicle development and may be a predictor of IVF outcome. PMID- 15890734 TI - Glucose intolerance, insulin resistance and cardiovascular risk factors in first degree relatives of women with polycystic ovary syndrome. AB - BACKGROUND: The aim of the present study was to evaluate insulin resistance (IR), glucose tolerance status and cardiovascular risk factors in first degree relatives of patients with polycystic ovary syndrome (PCOS). METHODS: A total of 120 family members [Mothers(PCOS) (n = 40), Fathers(PCOS) (n = 38), Sisters(PCOS) (n = 25) and Brothers(PCOS) (n = 17)] of 55 patients with PCOS and 75 unrelated healthy control subjects without a family history of diabetes or PCOS (four age- and weight-matched subgroups, i.e. Control(Mothers), Control(Fathers), Control(Sisters) and Control(Brothers)) were studied. IR was assessed by homeostatic model assessment (HOMA IR), log HOMA, insulin sensivity index (ISI), the quantitative insulin sensitivity check index (QUICKI) and area under the curve for insulin during the oral glucose tolerance test (AUCI, AUCG) in with normal glucose tolerance (NGT) subjects and controls. Serum adiponectin, resistin, homocysteine and lipid levels were measured. RESULTS: The prevalence of any degree of glucose intolerance was 40% in Mothers(PCOS) and 52% in Fathers(PCOS). In total, six (15%) glucose tolerance disorders were identified in the Control(Mothers) and Control(Fathers) in first degree relatives of control subjects. The first degree relatives of PCOS patients had significantly higher serum fasting insulin, HOMA-IR, Log HOMA and AUCI levels in all subgroups than the control subjects. The control subjects had significantly elevated QUCKI, ISI levels and serum adiponectin levels compared to the first degree relatives of PCOS subjects in all subgroups. The serum Hcy and resistin levels increased significantly in both Fathers(PCOS) and Mothers(PCOS) groups but not Brothers(PCOS) and Sister(PCOS). CONCLUSION: The results of the present study support the finding that the first degree relatives of PCOS patients carry an increased risk of cardiovascular disease, as do PCOS patients. PMID- 15890735 TI - Prolongation of follicular phase by delaying hCG administration results in a higher incidence of endometrial advancement on the day of oocyte retrieval in GnRH antagonist cycles. AB - BACKGROUND: Prolongation of follicular phase by delaying hCG administration has been reported to result in a significantly lower ongoing pregnancy rate that did not seem to be due to an embryonic factor. The aim of this prospective randomized study was to assess the effect of delaying hCG administration on endometrial histology. METHODS: Ten oocyte donors underwent endometrial biopsy on the day of oocyte retrieval and endometrial histology was assessed by Noyes' criteria. Ovarian stimulation was performed with recombinant (r)FSH and daily GnRH antagonist starting on day 6 of stimulation. Patients were randomized by a computer-generated list to receive 10 000 IU of hCG either as soon as > or =3 follicles > or =17 mm were present on ultrasound (early-hCG group, n = 5) or 2 days after this criterion was met (late-hCG group, n = 5). RESULTS: When hCG was delayed, endometrial advancement was present in all samples examined (median advancement 3 days, range 2-3 days). On the contrary, no secretory changes were observed when the follicular phase was not prolonged (difference in the proportion of patients with advancement between the early-hCG and the late-hCG group: 100%, 95% CI: 38-100). CONCLUSIONS: Prolongation of follicular phase by delaying hCG administration results in a higher incidence of endometrial advancement on the day of oocyte retrieval in GnRH antagonist cycles. PMID- 15890736 TI - Glutathione and free sulphydryl content of seminal plasma in healthy medical students during and after exam stress. AB - BACKGROUND: It has been reported that there is a relationship between stress and infertility. The mechanisms of stress-related semen quality alterations have not been fully elucidated. In the present study, we investigated the effect of examination stress on seminal glutathione and free sulphydryl content and sperm quality. METHODS: Semen samples were collected from 34 healthy volunteers who were students of medical school in the fourth semester just before (stress period) and 3 months after (non-stress period) their final examinations. Their psychological examination stress was measured by the State Trait Anxiety Inventory (STAI) questionnaire. After standard semen analysis, semen samples were centrifuged at 10 000g for 15 min. Glutathione and free sulphydryl concentration of seminal plasma were measured. RESULTS: During the period of examination stress, the glutathione and free sulphydryl content of seminal plasma and the motility index of spermatozoa were significantly lower, whereas the percentage of morphologically abnormal spermatozoa was higher, than during the non-stress period (P < 0.001, for all). An association between seminal plasma glutathione and motility index was observed at both periods (P < 0.05 and P < 0.01, respectively). CONCLUSIONS: This study demonstrated that glutathione and free sulphydryl levels in seminal plasma decreased in subjects undergoing examination stress. Furthermore, poor sperm quality may be due to loss of glutathione and free sulphydryl content of seminal plasma. PMID- 15890737 TI - Comparative study of plasma ghrelin levels in women with polycystic ovary syndrome, in hyperandrogenic women and in normal controls. AB - BACKGROUND: Ghrelin is a novel peptide associated with energy balance, obesity, and perhaps gonadal function. The present study was designed in order: (i) to compare plasma ghrelin levels between women with PCOS, women who presented only with hyperandrogenaemia and healthy controls; and (ii) to investigate the relationship between circulating ghrelin and the heterogeneity of clinical and biochemical manifestations of PCOS. METHODS: Two hundred and fifty-nine women with PCOS, 25 women who had only hyperandrogenaemia and 46 controls, were studied. Women with PCOS were further divided, based on the presence of chronic anovulation, biochemical hyperandrogenaemia, clinical hyperandrogenism, and polycystic ovary morphology on ultrasound evaluation. In all women, the basal levels of gonadotrophins, androgens, 17-OH-progesterone, sex hormone-binding globulin, glucose, insulin and ghrelin were measured. RESULTS: Women with PCOS had lower ghrelin levels, compared to both women with hyperandrogenaemia and controls; women with hyperandrogenaemia had lower ghrelin levels, compared to controls, but not significantly so. While PCOS-associated hyperandrogenaemia was inversely related to ghrelin levels, anovulation and polycystic ovary morphology were associated with higher concentrations. Ghrelin levels were negatively correlated with 17-OH-progesterone levels. CONCLUSIONS: In PCOS, circulating ghrelin and androgens are inversely related and it is possible that this peptide is involved in steroidal synthesis and/or action. It is also likely that different clinical and biochemical manifestations of the syndrome are also associated with different ghrelin concentrations. PMID- 15890738 TI - Season of birth influences the timing of menopause. AB - BACKGROUND: Seasons may influence prenatal growth and future fertility. This study investigated whether season and month of birth influenced the timing of menopause in a group of women attending three Italian menopause clinics. METHODS AND RESULTS: Age at menopause of 2822 post-menopausal women (>12 months of amenorrhoea) was stratified by month and season of birth. Mean age at menopause was 49.42 years (SEM: 0.78 years). Menopause occurred earlier for women born in the spring (age 49.04+/-0.15 years) than in the autumn (49.97+/-0.14 years). The earliest menopause was found in women born in March (48.9+/-0.25 years) and the latest in women born in October (50.3+/-0.25 years). The effect of season of birth on age at menopause remained even when considering factors that in our analysis were capable of significantly interfering with the timing of menopause, such as age at menarche, body mass index, smoking habit, level of education and type of job. CONCLUSIONS: Taking into consideration the retrospective design of the study, and a possible recall bias, the present data seem to suggest that environmental factors linked to seasons are capable of interfering with the timing of a woman's ovarian exhaustion by an action exerted in the prenatal period. PMID- 15890739 TI - Synapsis and meiotic recombination analyses: MLH1 focus in the XY pair as an indicator. AB - BACKGROUND: Anomalies in meiotic prophase I have been related to partial or total meiotic arrest. These anomalies include an abnormal synaptic process, resulting in disorders in meiotic recombination. METHODS: In the present study, we analyse primary spermatocytes from 12 infertile men (four with non-obstructive azoospermia, six with oligoastenoteratozoospermia, one with astenoteratozoospermia and one normozoospermic) and five control fertile donors using immunocytological techniques for synaptonemal complex, meiotic recombination and centromeric proteins. RESULTS: Mean numbers of MLH1 foci per cell, frequencies of cells presenting an MLH1 focus in the XY pair and percentages of cells affected by abnormal synaptic patterns (gaps and splits) are reported for each of the infertile patients and control men. A positive correlation between the frequency of cells showing a recombination focus in the XY pair and the number of autosomal recombination foci per cell is found. CONCLUSIONS: Reduced recombination in the XY pair and an increased number of cells affected by gaps may explain some idiopathic male infertility cases. The results suggest that recombination in the XY pair could be an indicator for general recombination frequency and for a successful meiotic process. PMID- 15890740 TI - GnRH agonist as novel luteal support: results of a randomized, parallel group, feasibility study using intranasal administration of buserelin. AB - BACKGROUND: The study objective was to investigate whether repeated intranasal administration of a GnRH agonist could provide convenient and safe luteal support. METHODS: Twenty-four patients with unexplained infertility were enrolled. All patients were treated with an aromatase inhibitor. When ovulation trigger criteria were met, patients were randomly allocated to either 5000 IU hCG (group A), or 200 microg intranasal buserelin followed by 100 microg every 3 days (group B), 100 microg every 2 days (group C), or 100 microg every day (group D), up to day 14 of the luteal phase. All patients underwent intrauterine insemination. RESULTS: Follicular development was similar in all groups with 1.1 +/- 0.3 follicles > or = 16 mm, 229.4 +/- 95.2 pg/ml estradiol (E2) and 0.8 +/- 0.5 ng/ml progesterone (mean+/-SD). The luteal phase duration (median; 95% confidence interval) was 15 (14.1, 15.0), 14 (12.5, 15.5), 15 (11.8, 18.2) and 15 (14.4, 15.6) days in groups A, B, C and D respectively. From luteal phase day 7 onwards, progesterone levels tended to be higher in group D compared with A. On day 14 of the luteal phase, progesterone levels were 3.0 (0.8, 5.2), 1.7 (-0.5, 3.9), 3.9 (-0.7, 8.5) and 7.7 (3.4, 11.9) ng/ml in groups A, B, C and D respectively (P = 0.045). No pregnancy was recorded in group A, but there was one biochemical pregnancy in group B, one biochemical and one singleton clinical pregnancy in group C, and two singleton clinical pregnancies in group D. CONCLUSION: Intranasal administration of buserelin could be effective to provide luteal support. This treatment was associated with a good pregnancy rate (5/18, 28%). PMID- 15890741 TI - Resolving methodological and clinical issues in the design of efficacy trials in assisted reproductive technologies: a mini-review. AB - The validity, importance and relevance of randomized controlled trials depend on identifying an appropriate target population, ensuring adequate power, careful attention to the details of randomization and blinding, and selection of an endpoint that is important to the target population. With efficacy trials more than effectiveness trials, additional constraints are needed to reduce the variability that is typical of clinical practice: a narrowly defined sample, unvarying pre-randomization procedures and post-randomization treatments and follow-up that are as identical as possible for all patients. Efficacy trials comparing ovarian stimulation protocols should have strict protocol definitions, specific concomitant medications and minimal variability between centres with respect to stimulation goals and dose adjustments. Additionally, there should be narrowly defined criteria for administration of chorionic gonadotrophin, type of luteal support, embryo transfer and freezing policies. The goal of efficacy trials is to minimize the variability that is extrinsic to the comparison. When efficacy has been proven, effectiveness trials are needed to determine whether the effect of the new intervention is robust in the variability of typical clinical settings. PMID- 15890742 TI - M@CBETH: a microarray classification benchmarking tool. AB - Microarray classification can be useful to support clinical management decisions for individual patients in, for example, oncology. However, comparing classifiers and selecting the best for each microarray dataset can be a tedious and non straightforward task. The M@CBETH (a MicroArray Classification BEnchmarking Tool on a Host server) web service offers the microarray community a simple tool for making optimal two-class predictions. M@CBETH aims at finding the best prediction among different classification methods by using randomizations of the benchmarking dataset. The M@CBETH web service intends to introduce an optimal use of clinical microarray data classification. PMID- 15890743 TI - Proteomic mass spectra classification using decision tree based ensemble methods. AB - MOTIVATION: Modern mass spectrometry allows the determination of proteomic fingerprints of body fluids like serum, saliva or urine. These measurements can be used in many medical applications in order to diagnose the current state or predict the evolution of a disease. Recent developments in machine learning allow one to exploit such datasets, characterized by small numbers of very high dimensional samples. RESULTS: We propose a systematic approach based on decision tree ensemble methods, which is used to automatically determine proteomic biomarkers and predictive models. The approach is validated on two datasets of surface-enhanced laser desorption/ionization time of flight measurements, for the diagnosis of rheumatoid arthritis and inflammatory bowel diseases. The results suggest that the methodology can handle a broad class of similar problems. PMID- 15890744 TI - Discovering patterns to extract protein-protein interactions from the literature: Part II. AB - MOTIVATION: An enormous number of protein-protein interaction relationships are buried in millions of research articles published over the years, and the number is growing. Rediscovering them automatically is a challenging bioinformatics task. Solutions to this problem also reach far beyond bioinformatics. RESULTS: We study a new approach that involves automatically discovering English expression patterns, optimizing them and using them to extract protein-protein interactions. In a sister paper, we described how to generate English expression patterns related to protein-protein interactions, and this approach alone has already achieved precision and recall rates significantly higher than those of other automatic systems. This paper continues to present our theory, focusing on how to improve the patterns. A minimum description length (MDL)-based pattern optimization algorithm is designed to reduce and merge patterns. This has significantly increased generalization power, and hence the recall and precision rates, as confirmed by our experiments. AVAILABILITY: http://spies.cs.tsinghua.edu.cn. PMID- 15890745 TI - A criticality-based framework for task composition in multi-agent bioinformatics integration systems. AB - MOTIVATION: During task composition, such as can be found in distributed query processing, workflow systems and AI planning, decisions have to be made by the system and possibly by users with respect to how a given problem should be solved. Although there is often more than one correct way of solving a given problem, these multiple solutions do not necessarily lead to the same result. Some researchers are addressing this problem by providing data provenance information. Others use expert advice encoded in a supporting knowledge-base. In this paper, we propose an approach that assesses the importance of such decisions with respect to the overall result. We present a way of measuring decision criticality and describe its potential use. RESULTS: A multi-agent bioinformatics integration system is used as the basis of a framework that facilitates such functionality. We propose an agent architecture, and a concrete bioinformatics example (prototype) is used to show how certain decisions may not be critical in the context of more complex tasks. PMID- 15890746 TI - Cis-regulatory element based targeted gene finding: genome-wide identification of abscisic acid- and abiotic stress-responsive genes in Arabidopsis thaliana. AB - MOTIVATION: A fundamental problem of computational genomics is identifying the genes that respond to certain endogenous cues and environmental stimuli. This problem can be referred to as targeted gene finding. Since gene regulation is mainly determined by the binding of transcription factors and cis-regulatory DNA sequences, most existing gene annotation methods, which exploit the conservation of open reading frames, are not effective in finding target genes. RESULTS: A viable approach to targeted gene finding is to exploit the cis-regulatory elements that are known to be responsible for the transcription of target genes. Given such cis-elements, putative target genes whose promoters contain the elements can be identified. As a case study, we apply the above approach to predict the genes in model plant Arabidopsis thaliana which are inducible by a phytohormone, abscisic acid (ABA), and abiotic stress, such as drought, cold and salinity. We first construct and analyze two ABA specific cis-elements, ABA responsive element (ABRE) and its coupling element (CE), in A.thaliana, based on their conservation in rice and other cereal plants. We then use the ABRE-CE module to identify putative ABA-responsive genes in A.thaliana. Based on RT-PCR verification and the results from literature, this method has an accuracy rate of 67.5% for the top 40 predictions. The cis-element based targeted gene finding approach is expected to be widely applicable since a large number of cis-elements in many species are available. PMID- 15890747 TI - ANOVA-simultaneous component analysis (ASCA): a new tool for analyzing designed metabolomics data. AB - MOTIVATION: Datasets resulting from metabolomics or metabolic profiling experiments are becoming increasingly complex. Such datasets may contain underlying factors, such as time (time-resolved or longitudinal measurements), doses or combinations thereof. Currently used biostatistics methods do not take the structure of such complex datasets into account. However, incorporating this structure into the data analysis is important for understanding the biological information in these datasets. RESULTS: We describe ASCA, a new method that can deal with complex multivariate datasets containing an underlying experimental design, such as metabolomics datasets. It is a direct generalization of analysis of variance (ANOVA) for univariate data to the multivariate case. The method allows for easy interpretation of the variation induced by the different factors of the design. The method is illustrated with a dataset from a metabolomics experiment with time and dose factors. PMID- 15890748 TI - PROFcon: novel prediction of long-range contacts. AB - MOTIVATION: Despite the continuing advance in the experimental determination of protein structures, the gap between the number of known protein sequences and structures continues to increase. Prediction methods can bridge this sequence structure gap only partially. Better predictions of non-local contacts between residues could improve comparative modeling, fold recognition and could assist in the experimental structure determination. RESULTS: Here, we introduced PROFcon, a novel contact prediction method that combines information from alignments, from predictions of secondary structure and solvent accessibility, from the region between two residues and from the average properties of the entire protein. In contrast to some other methods, PROFcon predicted short and long proteins at similar levels of accuracy. As expected, PROFcon was clearly less accurate when tested on sparse evolutionary profiles, that is, on families with few homologs. Prediction accuracy was highest for proteins belonging to the SCOP alpha/beta class. PROFcon compared favorably with state-of-the-art prediction methods at the CASP6 meeting. While the performance may still be perceived as low, our method clearly pushed the mark higher. Furthermore, predictions are already accurate enough to seed predictions of global features of protein structure. PMID- 15890749 TI - Motor skill training and strength training are associated with different plastic changes in the central nervous system. AB - Changes in corticospinal excitability induced by 4 wk of heavy strength training or visuomotor skill learning were investigated in 24 healthy human subjects. Measurements of the input-output relation for biceps brachii motor evoked potentials (MEPs) elicited by transcranial magnetic stimulation were obtained at rest and during voluntary contraction in the course of the training. The training paradigms induced specific changes in the motor performance capacity of the subjects. The strength training group increased maximal dynamic and isometric muscle strength by 31% (P < 0.001) and 12.5% (P = 0.045), respectively. The skill learning group improved skill performance significantly (P < 0.001). With one training bout, the only significant change in transcranial magnetic stimulation parameters was an increase in skill learning group maximal MEP level (MEP(max)) at rest (P = 0.02) for subjects performing skill training. With repeated skill training three times per week for 4 wk, MEP(max) increased and the minimal stimulation intensity required to elicit MEPs decreased significantly at rest and during contraction (P < 0.05). In contrast, MEP(max) and the slope of the input output relation both decreased significantly at rest but not during contraction in the strength-trained subjects (P < or = 0.01). No significant changes were observed in a control group. A significant correlation between changes in neurophysiological parameters and motor performance was observed for skill learning but not strength training. The data show that increased corticospinal excitability may develop over several weeks of skill training and indicate that these changes may be of importance for task acquisition. Because strength training was not accompanied by similar changes, the data suggest that different adaptive changes are involved in neural adaptation to strength training. PMID- 15890750 TI - Muscle activation and its distribution within human triceps surae muscles. AB - The purposes of this study were 1) to quantify the volume of activated parts within a whole muscle and 2) to examine activated area distributions along the length of muscle. Seven male subjects performed five sets of 10 repetitions of a single-leg calf-raise exercise with the knee fully extended. Transverse relaxation time (T2)-weighted spin echo images were acquired before and immediately after the exercise. A range of pixels with a T2 greater than the mean +1 SD of the region of interest (ROI) from the preexercise image and pixels with a T2 lower than the mean + SD of the ROI from the postexercise image were defined as "active" muscle. The active muscle images were three dimensionally reconstructed, from which the volume of the activated muscle was determined for individual triceps surae (TS) muscles. Our data indicate that approximately 46% of the medial gastrocnemius (MG) muscle was activated during the exercise, with activation of the lateral gastrocnemius (LG) and soleus (Sol) muscles being approximately 35%. In the MG, distal portions had a greater percentage area of activated muscle than the proximal portions (P < 0.05), which was consistent with the results regarding electromyogram activity. In contrast, regional activation differences were not observed in the LG and Sol. These findings suggest that the amounts of activated muscle and its distribution would be different among TS muscles. PMID- 15890751 TI - Reactive oxidant and p42/44 MAP kinase signaling is necessary for mechanical strain-induced proliferation in pulmonary epithelial cells. AB - Mechanical strain is necessary for normal lung growth and development. Individuals with respiratory failure are supported with mechanical ventilation, leading to altered lung growth and injury. Understanding signaling pathways initiated by mechanical strain in lung epithelial cells will help guide development of strategies aimed at optimizing strain-induced lung growth while mitigating ventilator-induced lung injury. To study strain-induced proliferative signaling, focusing on the role of reactive oxidant species (ROS) and p42/44 mitogen-activated protein (MAP) kinase, human pulmonary epithelial H441 and MLE15 cells were exposed to equibiaxial cyclic mechanical strain. ROS were increased within 15 min of strain. N-acetylcysteine inactivated strain-induced ROS and inhibited p42/44 MAP kinase phosphorylation and strain-induced proliferation. PD98059 and UO126, p42/44 MAP kinase inhibitors, blocked strain-induced proliferation. To verify the specificity of p42/44 MAP kinase inhibition, cells were transfected with dominant-negative mitogen-activated protein kinase kinase-1 plasmid DNA. Transfected cells did not proliferate in response to mechanical strain. To determine whether strain-induced tyrosine kinase activity is necessary for strain-induced ROS-p42/44 MAP kinase signaling, genistein, a tyrosine kinase inhibitor, was used. Genistein did not block strain-induced ROS production or p42/44 MAP kinase phosphorylation. Gadolinium, a mechanosensitive calcium channel blocker, blocked strain-induced ROS production and p42/44 MAP kinase phosphorylation but not strain-induced tyrosine phosphorylation. These data support ROS production and p42/44 MAP kinase phosphorylation being involved in a common strain-induced signaling pathway, necessary for strain-induced proliferation in pulmonary epithelial cells, with a parallel strain-induced tyrosine kinase pathway. PMID- 15890752 TI - Subcutaneous PO2 as an index of the physiological limits for hemodilution in the rat. AB - This study was designed to test the hypothesis that changes in subcutaneous PO2 (PscO2) during progressive hemodilution will reliably predict a "critical point" at which tissue O2 consumption (VO2) becomes dependent on O2 delivery (QO2). Twelve pentobarbital-anesthetized male Sprague-Dawley rats (315-375 g) underwent stepwise exchange of plasma for blood (1.5 ml of plasma for each 1 ml of blood lost). The initial exchange was equal to 25% of the estimated circulatory blood volume, and each subsequent exchange was equal to 10% of the estimated circulatory blood volume. After nine exchanges, the hematocrit (Hct) fell from 42 +/- 1 to 6 +/- 1%. Cardiac output and O2 extraction rose significantly. PscO2 became significantly reduced (P < 0.05) after exchange of 45% of the blood volume (Hct = 16 +/- 1%). VO2 became delivery dependent when QO2 fell below 21 ml x min( 1) x kg body wt(-1) (mean Hct = 13 +/- 1%). Eight control rats undergoing 1:1 blood-blood exchange showed no change in PscO2, pH, HCO3(-), or hemodynamics. Measurement of PscO2 may be a useful guide to monitor the adequacy of QO2 during hemodilution. PMID- 15890753 TI - Elevated temperature decreases sensitivity of P2X purinergic receptors in skeletal muscle arteries. AB - We hypothesized that elevated temperatures would attenuate but that reduced temperatures would potentiate the tension mediated by vascular P2X purinergic receptors. The femoral arteries of 24 rats were dissected out and placed in modified Krebs-Henseleit buffer. Arteries were cut into 2-mm sections and mounted in organ tissue baths. Maximal tension (g) was measured during a KCl and norepinephrine challenge. Tension was measured during doses of alpha,beta methylene ATP (10(-7) to 10(-3) M), phenylephrine (10(-7) to 10(-4) M), and acetylcholine (10(-9) to 10(-5) M), with tissue bath temperature adjusted to 35, 37, and 41 degrees C. Dose-response curves were fit using nonlinear regression analysis to calculate the EC50 and slope. The peak tension was lower with alpha,beta-methylene ATP during 41 degrees C (1.49 +/- 0.14 g) compared with 35 degrees C (2.08 +/- 0.09 g) and 37 degrees C (1.94 +/- 0.09 g; P < 0.05). Slope and EC50 were not affected by temperature. Tension produced by phenylephrine and relaxation to acetylcholine were not affected by temperature. These data indicate that the vasoconstrictor response to alpha,beta-methylene ATP is sensitive to temperature. Moderate cooling does not potentiate P2X-mediated vasoconstriction, but elevated temperature attenuates the vasoconstrictor response to P2X purinergic receptors. PMID- 15890754 TI - Induction of long-lasting depolarization in medioventral medulla neurons by cholinergic input from the pedunculopontine nucleus. AB - Stimulation of the pedunculopontine nucleus (PPN) is known to induce changes in arousal and postural/locomotor states by activation of such descending targets as the caudal pons and the medioventral medulla (MED). Previously, PPN stimulation was reported to induce prolonged responses (PRs) in intracellularly recorded caudal pontine neurons in vitro. The present study used intracellular recordings in semihorizontal slices from rat brain stem (postnatal days 12-21) to determine responses in MED neurons following PPN stimulation. One-half (40/81) of MED neurons showed PRs after PPN stimulation. MED neurons with PRs had shorter duration action potential, longer duration afterhyperpolarization, and higher amplitude afterhyperpolarization than non-PR MED neurons. PR MED neurons were significantly larger (568 +/- 44 microm2) than non-PR MED neurons (387 +/- 32 microm2). The longest mean duration PRs and maximal firing rates during PRs were induced by PPN stimulation at 60 Hz compared with 10, 30, or 90 Hz. The muscarinic cholinergic agonist carbachol induced depolarization in all PR neurons tested, and the muscarinic cholinergic antagonist scopolamine reduced or blocked carbachol- and PPN stimulation-induced PRs in all MED neurons tested. These findings suggest that PPN stimulation-induced PRs may be due to activation of muscarinic receptor-sensitive channels, allowing MED neurons to respond to a transient, frequency-dependent depolarization with long-lasting stable states. PPN stimulation appears to induce PRs in large MED neurons using parameters known best to induce locomotion. PMID- 15890755 TI - Red blood cell lactate transport in sickle disease and sickle cell trait. AB - This study determined and compared rates and mechanisms of lactate transport in red blood cells (RBCs) of persons with 1) sickle cell disease (HbSS), 2) sickle cell trait (HbAS), and 3) a control group (HbAA). Blood samples were drawn from 30 African-American volunteers (10 HbSS, 10 HbAS, 10 HbAA). Lactate influx into RBCs was measured by using [14C]lactate at six (2, 5, 10, 15, 25, and 40 mM) unlabeled lactate concentrations. The monocarboxylate transporter pathway was blocked by p-chloromercuriphenylsulfonic acid to determine its percent contribution to total lactate influx. Generally, total lactate influx into RBCs from the HbSS group was significantly greater than influx into RBCs from HbAS or HbAA, with no difference between HbAS and HbAA. Faster influx into HbSS RBCs was attributed to increased monocarboxylate transporter activity [increased apparent Vmax (V'max)]. V'max (4.7 +/- 0.6 micromol x ml(-1) x min(-1)) for HbSS RBCs was significantly greater than V'max of HbAS RBCs (2.9 +/- 1.5 micromol x ml(-1) x min(-1)) and HbAA RBCs (2.0 +/- 0.5 micromol x ml(-1) x min(-1)). Km (42.8 +/- 8 mM) for HbSS RBCs was significantly greater than Km (27 +/- 12 mM) for HbAA RBCs. We suspect that elevated erythropoietin levels in response to chronic anemia and/or pharmacological treatment (erythropoietin injections, hydroxyurea ingestion) is the underlying mechanism for increased lactate transport capacity in HbSS RBCs. PMID- 15890756 TI - Effects of prior heavy-intensity exercise during single-leg knee extension on VO2 kinetics and limb blood flow. AB - The effects of prior heavy-intensity exercise on O(2) uptake (Vo(2)) kinetics of a second heavy exercise may be due to vasodilation (associated with metabolic acidosis) and improved muscle blood flow. This study examined the effect of prior heavy-intensity exercise on femoral artery blood flow (Qleg) and its relationship with Vo(2) kinetics. Five young subjects completed five to eight repeats of two 6 min bouts of heavy-intensity one-legged, knee-extension exercise separated by 6 min of loadless exercise. Vo(2) was measured breath by breath. Pulsed-wave Doppler ultrasound was used to measure Qleg. Vo(2) and blood flow velocity data were fit using a monoexponential model to identify phase II and phase III time periods and estimate the response amplitudes and time constants (tau). Phase II Vo(2) kinetics was speeded on the second heavy-intensity exercise [mean tau (SD), 29 (10) s to 24 (10) s, P < 0.05] with no change in the phase II (or phase III) amplitude. Qleg was elevated before the second exercise [1.55 (0.34) l/min to 1.90 (0.25) l/min, P < 0.05], but the amplitude and time course [tau, 25 (13) s to 35 (13) s] were not changed, such that throughout the transient the Qleg (and DeltaQleg/DeltaVo(2)) did not differ from the prior heavy exercise. Thus Vo(2) kinetics were accelerated on the second exercise, but the faster kinetics were not associated with changes in Qleg. Thus limb blood flow appears not to limit Vo(2) kinetics during single-leg heavy-intensity exercise nor to be the mechanism of the altered Vo(2) response after heavy-intensity prior exercise. PMID- 15890757 TI - Marching to the beat of the same drummer: the spontaneous tempo of human locomotion. AB - Laboratory studies have suggested that the preferred cadence of walking is approximately 120 steps/min, and the vertical acceleration of the head exhibits a dominant peak at this step frequency (2 Hz). These studies have been limited to short periods of walking along a predetermined path or on a treadmill, and whether such a highly tuned frequency of movement can be generalized to all forms of locomotion in a natural setting is unknown. The aim of this study was to determine whether humans exhibit a preferred cadence during extended periods of uninhibited locomotor activity and whether this step frequency is consistent with that observed in laboratory studies. Head linear acceleration was measured over a 10-h period in 20 subjects during the course of a day, which encompassed a broad range of locomotor (walking, running, cycling) and nonlocomotor (working at a desk, driving a car, riding a bus or subway) activities. Here we show a highly tuned resonant frequency of human locomotion at 2 Hz (SD 0.13) with no evidence of correlation with gender, age, height, weight, or body mass index. This frequency did not differ significantly from the preferred step frequency observed in the seminal laboratory study of Murray et al. (Murray MP, Drought AB, and Kory RC. J Bone Joint Surg 46A: 335-360, 1964). [1.95 Hz (SD 0.19)]. On the basis of the frequency characteristics of otolith-spinal reflexes, which drive lower body movement via the lateral vestibulospinal tract, and otolith-mediated collic and ocular reflexes that maintain gaze when walking, we speculate that this spontaneous tempo of locomotion represents some form of central "resonant frequency" of human movement. PMID- 15890758 TI - Dose-response relationship of endurance training for autonomic circulatory control in healthy seniors. AB - Aging results in marked abnormalities of cardiovascular regulation. Regular exercise can improve many of these age-related abnormalities. However, it remains unclear how much exercise is optimal to achieve this improvement or whether the elderly can ever improve autonomic control by exercise training to a degree similar to that observed in healthy young individuals. Ten healthy sedentary seniors [71 +/- 3 (SD) yr] trained for 12 mo; training involved progressive increases in volume and intensity. Static hemodynamics were measured, and R-wave R-wave interval (RRI), beat-to-beat blood pressure (BP) variability, and transfer function gain between systolic BP and RRI were calculated at baseline and every 3 mo during training. Data were compared with those obtained in 12 Masters athletes (68 +/- 3 yr) and 11 healthy sedentary young individuals (29 +/- 6 yr) at baseline. Additionally, the adaptation of these variables after completion of identical training loads was compared between the seniors and the young. Indexes of RRI variability and baroreflex gain were decreased in the sedentary seniors but preserved in the Masters athletes compared with the young at baseline. With training in the seniors, baroreflex gain and resting BP showed a peak adaptation after moderate doses of training following 3-6 mo. Indexes of RRI variability continued to improve with increasing doses of training and increased to the same magnitude as the young at baseline after heavy doses of training for 12 mo; however, baroreflex gain never achieved values equivalent to the young at baseline, even after a year of training. The magnitude of the adaptation of these variables to identical training loads was similar (no interaction effects of age x training) between the seniors and the young. Thus RRI variability in seniors improves with increasing "dose" of exercise over 1 yr of training. In contrast, more moderate doses of training for 3-6 mo may optimally improve baroreflex sensitivity, associated with a modest hypotensive effect; however, higher doses of training do not lead to greater enhancement of these changes. Seniors retain a similar degree of "trainability" as young subjects for cardiac autonomic function to dynamic exercise. PMID- 15890759 TI - Positive end-expiratory pressure differentially alters pulmonary hemodynamics and oxygenation in ventilated, very premature lambs. AB - In mature lungs, elevated positive end-expiratory pressure (PEEP) reduces pulmonary blood flow (PBF) and increases pulmonary vascular resistance (PVR). However, the effect of PEEP on PBF in preterm infants with immature lungs and a patent ductus arteriosus is unknown. Fetal sheep were catheterized at 124 days of gestation (term approximately 147 days), and a flow probe was placed around the left pulmonary artery to measure PBF. At 127 days, lambs were delivered and ventilated from birth with a tidal volume of 5 ml/kg and 4-cmH(2)O PEEP; PEEP was changed to 0, 8, and 12 cmH(2)O in random order, returning to 4 cmH(2)O between each change. Increasing PEEP from 4 to 8 cmH(2)O and from 4 to 12 cmH(2)O decreased PBF by 20.5 and 41.0%, respectively, and caused corresponding changes in PVR; reducing PEEP from 4 to 0 cmH(2)O did not affect PBF. Despite decreasing PBF, increasing PEEP from 4 to 8 cmH(2)O and 12 cmH(2)O improved oxygenation of lambs. Increasing and decreasing PEEP from 4 cmH(2)O significantly changed the contour of the PBF waveform; at a PEEP of 12 cmH(2)O, end-diastolic flow was reduced by 82.8% and retrograde flow was reestablished. Although increasing PEEP improves oxygenation, it adversely affects PBF and PVR shortly after birth, alters the PBF waveform, and reestablishes retrograde flow during diastole. PMID- 15890760 TI - Heart rate variability responses to hypoxic and hypercapnic exposures in different mouse strains. AB - Heart rate variability (HRV) is a well-characterized, noninvasive means of assessing cardiac autonomic nervous system activity. This study examines the basic cardiac responses to hypoxic and hypercapnic challenges in seven strains of commonly used inbred mice (A/J, BALB/cJ, C3H/HeJ, C57BL/6J, CBA/J, DBA/2J, and FVB/J). Adult male mice, 8-12 wk of age, were chronically instrumented to a femoral artery catheter for the continuous measurement of systemic arterial blood pressure and heart rate. Mice were exposed to multiple 4-min periods of hypoxia (10% O2), hypercapnia (5% CO2), and combined hypoxia/hypercapnia (10% O2 + 5% CO2). HRV was derived from pulse intervals of the blood pressure tracings. Hypoxia induced increases in high-frequency HRV power and decreased low-frequency (LF) HRV power in most strains. Hypercapnia led to decreased high-frequency HRV power and increased LF HRV power in most strains. Strain differences were most notable in regard to the concomitant exposures of hypoxia and hypercapnia, with FVB/J mice mirroring their own response to hypercapnia alone, whereas CBA/J mice mirrored their own responses to hypoxia. As blood pressure is most likely the driving factor for heart rate changes via the baroreflex pathway, it is interesting that LF, considered to reflect cardiac sympathetic activity, was negatively correlated with heart rate, suggesting that LF changes are driven by baroreflex oscillation and not necessarily by absolute sympathetic or parasympathetic activity to the heart. These findings suggest that genetic background can influence the centrally mediated cardiovascular responses to basic hypoxic and hypercapnic challenges. PMID- 15890761 TI - Shortchanged? An assessment of chronic disease programming in major US city health departments. AB - A self-administered survey was distributed to members of The Big Cities Health Coalition, a group of Health Officers/Commissioners from 17 of the largest US metropolitan health departments. The survey asked participants about their chronic disease priorities, data sources, budgets, and funding sources as well as examples of successful chronic disease interventions. Members of the Coalition discussed the survey results in a scheduled conference call. Chronic diseases account for 70% of all deaths nationwide on average, yet the health departments surveyed allocated an average of 1.85% of their budgets to chronic disease. Average chronic disease spending per inhabitant was 2.33 dollars, with a median of 1.56 dollars. Among the group's top chronic disease priorities were asthma, diabetes, tobacco, cancer, and cardiovascular disease (CVD). Nearly half of the group's chronic disease spending was on tobacco. Chronic disease funding sources varied across localities, but direct federal funding was minimal. In 14 cities serving a combined 37 million people (13% of the US population), direct federal chronic disease funding totaled 8.7 million dollars, an average of 0.24 dollars per capita. The group described successful chronic disease interventions, particularly related to tobacco and asthma. PMID- 15890762 TI - Disseminated coccidioidomycosis in an immunocompetent person living in New York City. AB - Coccidioidomycosis is a disease caused by Coccidioides immitis, a soil-inhabiting fungus endemic to the desert climate of the southwestern United States and Central and South America. We report a case of disseminated coccidioidomycosis in a previously healthy person living in New York City, who was initially thought to have tuberculosis. The incidence of coccidioidomycosis has been increasing in both endemic and nonendemic areas, but diagnosis is often delayed or missed in nonendemic areas, resulting in extensive and unnecessary medical workup for other diseases or progression to serious disease. Therefore, clinicians should increase their awareness and consideration of this disease in patients with chronic systemic illness. PMID- 15890763 TI - Improved quantification of left ventricular mass based on endocardial and epicardial surface detection with real time three dimensional echocardiography. AB - OBJECTIVE: To develop a technique for volumetric analysis of real time three dimensional echocardiography (RT3DE) data aimed at quantifying left ventricular (LV) mass and to validate the technique against magnetic resonance (MR) assumed as the reference standard. DESIGN: RT3DE, which has recently become widely available, provides dynamic pyramidal data structures that encompass the entire heart and allows four dimensional assessment of cardiac anatomy and function. However, analysis techniques for the quantification of LV mass from RT3DE data are fundamentally two dimensional, rely on geometric modelling, and do not fully exploit the volumetric information contained in RT3DE datasets. Twenty one patients underwent two dimensional echocardiography (2DE), RT3DE, and cardiac MR. LV mass was measured from 2DE and MR images by conventional techniques. RT3DE data were analysed to semiautomatically detect endocardial and epicardial LV surfaces by the level set approach. From the detected surfaces, LV mass was computed directly in the three dimensional space as voxel counts. RESULTS: RT3DE measurement was feasible in 19 of 21 patients and resulted in higher correlation with MR (r = 0.96) than did 2DE (r = 0.79). RT3DE measurements also had a significantly smaller bias (-2.1 g) and tighter limits of agreement (2SD = +/-23 g) with MR than did the 2DE values (bias (2SD) -34.9 (50) g). Additionally, interobserver variability of RT3DE (12.5%) was significantly lower than that of 2DE (24.1%). CONCLUSIONS: Direct three dimensional model independent LV mass measurement from RT3DE images is feasible in the clinical setting and provides fast and accurate assessment of LV mass, superior to the two dimensional analysis techniques. PMID- 15890764 TI - Primary percutaneous coronary intervention for patients with acute ST elevation myocardial infarction with and without diabetes mellitus. PMID- 15890765 TI - Depression after myocardial infarction is a risk factor for declining health related quality of life and increased disability and cardiac complaints at 12 months. AB - OBJECTIVE: To study the prospective effects of post-myocardial infarction (MI) depressive disorder on health status, including self reported cardiac symptoms, disability, and health related quality of life, at 12 months after MI while controlling for cardiac condition and health status at three months after MI. METHODS: Longitudinal study of a cohort of 468 adults with MI recruited from four hospitals in the north of The Netherlands between September 1997 and September 2000. Assessment of depressive disorder in the year after MI according to International classification of disease, 10th revision criteria. Assessment of health status at three and 12 months by the RAND 36 item health survey, the health complaints scale, self rated cardiac complaints, and disability. RESULTS: Patients with post-MI depressive disorder were more likely than those without to have poor quality of life, more health complaints, more cardiac complaints, and more disability at 12 months' follow up. In multivariate analyses adjusted for cardiac condition, health status at three months, age, sex, and pre-MI depression, the prospective association of post-MI depression with poor health status remained for most of the indicators. Severity of the post-MI depression further contributed to aspects of poor health status. CONCLUSIONS: In a prospective study design, post-MI depression had strong effects on poor health status exceeding the effects of cardiac condition and its short term consequences. Efforts to improve health status after MI should therefore include standard assessment and guideline based treatment of post-MI depressive disorder. PMID- 15890766 TI - Effect of alpha linolenic acid on cardiovascular risk markers: a systematic review. AB - OBJECTIVE: To determine whether dietary supplementation with alpha linolenic acid (ALA) can modify established and emerging cardiovascular risk markers. DESIGN: Systematic review and meta-analysis of randomised controlled trials identified by a search of Medline, Embase, Cochrane Controlled Trials Register (CENTRAL), and the metaRegister of Controlled Trials (mRCT). PATIENTS: All human studies were reviewed. MAIN OUTCOME MEASURES: Changes in concentrations of total cholesterol, low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, very low density lipoprotein (VLDL) cholesterol, triglyceride, fibrinogen, and fasting plasma glucose, and changes in body mass index, weight, and systolic and diastolic blood pressure. RESULTS: 14 studies with minimum treatment duration of four weeks were reviewed. ALA had a significant effect on three of the 32 outcomes examined in these studies. Concentrations of fibrinogen (0.17 micromol/l, 95% confidence interval (CI) -0.30 to -0.04, p = 0.01) and fasting plasma glucose (0.20 mmol/l, 95% CI -0.30 to -0.10, p < 0.01) were reduced. There was a small but clinically unimportant decrease in HDL (0.01 mmol/l, 95% CI -0.02 to 0.00, p < 0.01). Treatment with ALA did not significantly modify total cholesterol, triglycerides, weight, body mass index, LDL, diastolic blood pressure, systolic blood pressure, VLDL, and apolipoprotein B. CONCLUSIONS: Although ALA supplementation may cause small decreases in fibrinogen concentrations and fasting plasma glucose, most cardiovascular risk markers do not appear to be affected. Further trials are needed, but dietary supplementation with ALA to reduce cardiovascular disease cannot be recommended. PMID- 15890767 TI - C reactive protein concentration and recurrence of atrial fibrillation after electrical cardioversion. AB - BACKGROUND: To test the hypothesis that a high C reactive protein (CRP) concentration would predict recurrence of atrial fibrillation (AF) after cardioversion in patients taking antiarrhythmic drugs. METHODS: 111 patients who underwent direct current cardioversion for symptomatic AF were enrolled. Blood was drawn for CRP determination before cardioversion on the same day. All patients were taking antiarrhythmic drugs before and after electrical cardioversion. RESULTS: After a mean follow up of 76 days, 75 patients had recurrence of AF. In univariate analysis, the median CRP concentration was significantly higher in patients with AF recurrence (3.95 mg/l v 1.81 mg/l, p = 0.002). Among the 55 patients with CRP in the upper 50th centile, 44 (80%) experienced recurrence of AF over a total follow up of 8.98 patient years, whereas among the 56 patients with CRP in the lower 50th centile, 31 (55%) experienced recurrence of AF over a total follow up of 14.3 patient years (p < 0.001). The adjusted hazard ratio comparing the upper 50th centile of CRP with the lower 50th centile of CRP was 2.0 (95% confidence interval 1.2 to 3.2, p = 0.007). CONCLUSIONS: CRP is independently associated with recurrence of AF after electrical cardioversion among patients taking antiarrhythmic drugs. These results suggest that inflammation may have a role in the pathogenesis of AF resistant to antiarrhythmic drugs. PMID- 15890768 TI - Methoxychlor disrupts uterine Hoxa10 gene expression. AB - Methoxychlor (MXC) is a pesticide that has adverse effects on reproductive capability in mice. MXC and its metabolites bind the estrogen receptor and function as endocrine disruptors. MXC diminishes the uterine decidual cell response, necessary for the support of pregnancy. Hoxa10 is an estrogen-regulated gene that is an essential mediator of the decidual response and necessary for pregnancy. Here we demonstrate that a mechanism by which MXC disrupts uterine function is by suppressing Hoxa10 expression. MXC treatment of mice produced a mild uterotropic response as measured by increased uterine weight and epithelial height. MXC treatment of uterine Ishikawa cells in vitro induced Hoxa10 expression. Estrogen receptor (ER) binding to the HOXA10 estrogen response element (ERE) was promoted by treatment with estradiol (E2); however, MXC disrupted E2/ER/ERE complex formation and gel shift. MXC alone allowed weak ER/ERE binding. In vivo MXC blocked the effect of E2 on Hoxa10 expression. Neonatal MXC treatment resulted in an immediate suppression and cellular restriction of Hoxa10 expression as well as a permanent generalized decrease in expression that persisted in the adult. MXC inhibited the expression of Hoxa10, a gene necessary for uterine development and function. One common mechanism by which endocrine disrupting chemicals produce lasting reproductive tract defects is through permanent alteration of developmental gene expression. PMID- 15890769 TI - Heterodimeric fly glycoprotein hormone-alpha2 (GPA2) and glycoprotein hormone beta5 (GPB5) activate fly leucine-rich repeat-containing G protein-coupled receptor-1 (DLGR1) and stimulation of human thyrotropin receptors by chimeric fly GPA2 and human GPB5. AB - Glycoprotein hormones play important roles in thyroid and gonadal function in vertebrates. The glycoprotein hormone alpha-subunit forms heterodimers with different beta-subunits to activate TSH or gonadotropin (LH and FSH) receptors. Recent genomic analyses allowed the identification of another alpha-subunit, GPA2, and another beta-subunit, GPB5, in human, capable of forming heterodimers to activate TSH receptors. Based on comparative genomic searches, we isolated the fly orthologs for human GPA2 and GPB5, each consisting of 10 cysteine residues likely involved in cystine-knot formation. RT-PCR analyses in Drosophila melanogaster demonstrated the expression of GPA2 and GPB5 at different developmental stages. Immunoblot analyses further showed that fly GPA2 and GPB5 subunit proteins are of approximately 16 kDa, and coexpression of these subunits yielded heterodimers. Purified recombinant fly GPA2/GPB5 heterodimers were found to be glycoproteins with N-linked glycosylated alpha-subunits and nonglycosylated beta-subunits, capable of stimulating cAMP production mediated by fly orphan receptor DLGR1 but not DLGR2. Although the fly GPA2/GPB5 heterodimers did not activate human TSH or gonadotropin receptors, chimeric fly GPA2/human GPB5 heterodimers stimulated human TSH receptors. These findings indicated that fly GPA2/GPB5 is a ligand for DLGR1, thus showing the ancient origin of this glycoprotein hormone-seven transmembrane receptor-G protein signaling system. The fly GPA2 also could form heterodimers with human GPB5 to activate human TSH receptors, indicating the evolutionary conservation of these genes and suggesting that the GPA2 subunit may serve as a scaffold for the beta-subunit to activate downstream G protein-mediated signaling. PMID- 15890770 TI - Transactivation of the parathyroid hormone promoter by specificity proteins and the nuclear factor Y complex. AB - We previously identified a highly conserved specificity protein 1 (Sp1) DNA element in mammalian PTH promoters that acted as an enhancer of gene transcription and bound Sp1 and Sp3 proteins present in parathyroid gland nuclear extracts. More recently, a nuclear factor (NF)-Y element (NF-Y(prox)) was also described by our group, which was located approximately 30 bp downstream from the Sp1 site in the human PTH (hPTH) promoter and by itself acted as a weak enhancer of gene transcription. We now report that Sp proteins and NF-Y can synergistically enhance transcription of a minimal hPTH promoter construct. Positioning of the Sp1 DNA element appears to be critical for this synergism because deviations of one half of a helical turn caused an approximate 60% decrease in transactivation. Finally, examination of the bovine PTH (bPTH) promoter also revealed Sp1/NF-Y synergism, in conjunction with the identification of an analogous NF-Y binding site similarly positioned downstream from the bPTH Sp1 element. In summary, synergistic transactivation of the hPTH and bPTH promoters is observed by Sp proteins and the NF-Y complex. The conservation of this transactivation in the human and bovine promoters suggests that this may be a principle means of enhancing PTH gene transcription. PMID- 15890771 TI - The Spot 14 protein is required for de novo lipid synthesis in the lactating mammary gland. AB - We generated a Spot 14 null mouse to assess the role of Spot 14 in de novo lipid synthesis and report the Spot 14 null mouse exhibits a phenotype in the lactating mammary gland. Spot 14 null pups nursed by Spot 14 null dams gain significantly less weight than wild-type pups nursed by wild-type dams. In contrast, Spot 14 null pups nursed by heterozygous dams show similar weight gain to wild-type littermates. We found the triglyceride content in Spot 14 null milk is significantly reduced. We demonstrate this reduction is the direct result of decreased de novo lipid synthesis in lactating mammary glands, corroborated by a marked reduction of medium-chain fatty acids in the triglyceride pool. Importantly, the reduced lipogenic rate is not associated with significant changes in the activities or mRNA of key lipogenic enzymes. Finally, we report the expression of a Spot 14-related gene in liver and adipose tissue, which is absent in the lactating mammary gland. We suggest that expression of both the Spot 14 and Spot 14-related proteins is required for maximum efficiency of de novo lipid synthesis in vivo and that these proteins impart a novel mechanism regulating de novo lipogenesis. PMID- 15890772 TI - Angiotensin II dilates bovine adrenal cortical arterioles: role of endothelial nitric oxide. AB - Adrenal steroidogenesis is modulated by humoral and neuronal factors and blood flow. Angiotensin II (AII) stimulates adrenal cortical aldosterone and cortisol production and medullary catecholamine release. However, AII regulation of adrenal vascular tone has not been characterized. We examined the effect of AII on diameters of cannulated bovine adrenal cortical arteries. Cortical arteries (average internal diameter = 230 microm) were constricted with U46619 and concentration-diameter responses to AII (10(-13) to 10(-8) mol/liter) were measured. In endothelium-intact arteries, AII induced dilations at low concentrations (maximum dilation = 25 +/- 6% at 10(-10) mol/liter) and constrictions at high concentrations (maximum constriction = 25 +/- 18% at 10(-8) mol/liter). AII constrictions were blocked by the angiotensin type 1 (AT1) receptor antagonist, losartan (10(-6) mol/liter). AII dilations were enhanced by losartan (maximal dilation = 48 +/- 8%), abolished by endothelial cell removal or N-nitro-L-arginine (L-NA, 3 x 10(-5) mol/liter) and inhibited by the angiotensin type 2 (AT2) receptor antagonist, PD123319 (10(-6) mol/liter, maximal dilation = 18 +/- 4%). In a 4,5-diaminofluorescein diacetate nitric oxide (NO) assay of isolated cortical arteries, AII stimulated NO production, which was abolished by PD123319, L-NA, or endothelial cell removal. Western immunoblot of arterial homogenates and endothelial and zona glomerulosa cell lysates revealed 48-kD and 50-kD bands corresponding to AT1 and AT2 receptors, respectively, in all three and a 140-kD band corresponding to endothelial NO synthase in endothelial cells and arteries. Our results demonstrate that AII stimulates adrenal cortical arterial dilation through endothelial cell AT2 receptor activation and NO release and AT1 receptor-dependent constriction. PMID- 15890773 TI - Arginine residue 155 in the second intracellular loop plays a critical role in rat melanin-concentrating hormone receptor 1 activation. AB - Melanin-concentrating hormone (MCH) receptor 1 (MCH1R) is a class A G protein coupled receptor. The MCH system has been linked to a variety of physiological functions, including the regulation of feeding and energy metabolism. We recently reported the importance of a dibasic motif in the membrane-proximal C-terminal region for MCH1R function. Here we reveal that an Arg residue in intracellular loop 2 of MCH1R plays a critical role in receptor function. We analyzed the roles of two distinct motifs, BBXXB and BXBB (in which B is a basic residue and X is a nonbasic residue), located in the three intracellular loops of MCH1R. Triple substitution mutants of intracellular loops 1 and 3 could still activate calcium mobilization, albeit with lower efficacy or potency. However, mutations in intracellular loop 2 led to a complete loss of induction of signal transduction without changing the high affinity constant (Kd) value. By analyzing a series of single-substitution mutants, a point mutation of Arg155 in intracellular loop 2 was found to be responsible for the signaling pathway elicited by MCH. In addition, substitution at positions corresponding to Arg155 in human MCH receptor 2 and rat somatostatin receptor 2 also markedly abolished their ligand-induced signaling capacities, indicating that this Arg is a recognition determinant in several G protein-coupled receptors. PMID- 15890774 TI - Localization and action of Dragon (repulsive guidance molecule b), a novel bone morphogenetic protein coreceptor, throughout the reproductive axis. AB - Bone morphogenetic proteins (BMPs) play important roles in reproduction including primordial germ cell formation, follicular development, spermatogenesis, and FSH secretion. Dragon, a recently identified glycosylphosphatidylinositol-anchored member of the repulsive guidance molecule family, is also a BMP coreceptor. In the present study, we determined the tissue and cellular localization of Dragon in reproductive organs using immunohistochemistry and in situ hybridization. Among reproductive organs, Dragon was expressed in testis, epididymis, ovary, uterus, and pituitary. In the testis of early postnatal mice, Dragon was found in gonocytes and spermatogonia, whereas in immature testes, Dragon was only weakly expressed in spermatogonia. Interestingly, pregnant mare serum gonadotropin treatment of immature mice robustly induced Dragon production in spermatocytes. In adult testis, Dragon was found in spermatocytes and round spermatids. In the ovary, Dragon was detected exclusively within oocytes and primarily those within secondary follicles. In the pituitary, Dragon-expressing cells overlapped FSH expressing cells. Dragon was also expressed in a number of cell lines originating from reproductive tissues including Ishikawa, Hela, LbetaT2, MCF-7, and JEG3 cells. Immunocytochemistry and gradient sucrose ultracentrifugation studies showed Dragon was localized in lipid rafts within the plasma membrane. In reproductive cell lines, Dragon expression enhanced signaling of exogenous BMP2 or BMP4. The present studies demonstrate that Dragon expression is dynamically regulated throughout the reproductive tract and that Dragon protein modulates BMP signaling in cells from reproductive tissues. The overlap between Dragon expression and the functional BMP signaling system suggests that Dragon may play a role in mammalian reproduction. PMID- 15890775 TI - Hypothalamic cocaine- and amphetamine-regulated transcript (CART) and agouti related protein (AgRP) neurons coexpress the NOP1 receptor and nociceptin alters CART and AgRP release. AB - Nociceptin or orphanin FQ (N/OFQ) and its receptor NOP1 are expressed in hypothalamic nuclei involved in energy homeostasis. N/OFQ administered by intracerebroventricular or arcuate nucleus (ARC) injection increases food intake in satiated rats. The mechanisms by which N/OFQ increases food intake are unknown. We hypothesized that N/OFQ may regulate hypothalamic neurons containing peptides involved in the control of food intake such as cocaine- and amphetamine regulated transcript (CART), alphaMSH, neuropeptide Y (NPY), and agouti-related protein (AgRP). We investigated the ability of N/OFQ to alter the release of CART, alphaMSH, NPY, and AgRP using ex vivo medial basal hypothalamic explants. Incubation of hypothalamic explants with N/OFQ (1, 10, 100 nM) resulted in significant changes in CART and AgRP release. One hundred nanomoles N/OFQ caused a 33% decrease in release of CART (55-102) immunoreactivity (IR) and increased release of AgRP-IR to 163% but produced no change in either alphaMSH-IR or NPY IR. Double immunocytochemistry/in situ hybridization demonstrated that CART-IR and NOP1 mRNA are colocalized throughout the hypothalamus, in particular in the paraventricular nucleus, lateral hypothalamus, zona incerta, and ARC, providing an anatomical basis for N/OFQ action on CART release. Dual in situ hybridization demonstrated that AgRP neurons in the ARC also express the NOP1 receptor. Our data suggest that nociceptin via the NOP1 receptor may increase food intake by decreasing the release of the anorectic peptide CART and increasing the release of the orexigenic peptide AgRP. PMID- 15890776 TI - Peripheral interaction of ghrelin with cholecystokinin on feeding regulation. AB - Ghrelin and cholecystokinin (CCK) are gastrointestinal hormones regulating feeding. Both transmitted via the vagal afferent, ghrelin elicits starvation signals, whereas CCK induces satiety signals. We investigated the interaction between ghrelin and CCK functioning in short-term regulation of feeding in Otsuka Long-Evans Tokushima fatty (OLETF) rats, which have a disrupted CCK type A receptor (CCK-AR), and their lean littermates, Long-Evans Tokushima Otsuka (LETO) rats. Intravenous administration of ghrelin increased 2-h food intake in both OLETF and LETO rats. Because OLETF rats are CCK insensitive, iv-administered CCK decreased 2-h food intake in LETO, but not in OLETF, rats. Although preadministration of CCK to LETO rats blocked food intake induced by ghrelin, CCK preadministration to OLETF rats did not affect ghrelin-induced food intake. Conversely, preadministration of ghrelin to LETO rats blocked feeding reductions induced by CCK. In electrophysiological studies, once gastric vagal afferent discharges were altered by ghrelin or CCK administration, they could not be additionally affected by serial administrations of either CCK or ghrelin, respectively. The induction of Fos expression in the hypothalamic arcuate nucleus by ghrelin was also attenuated by CCK preadministration. Using immunohistochemistry, we also demonstrated the colocalization of GH secretagogue receptor (GHS-R), the cellular receptor for ghrelin, with CCK-AR in vagal afferent neurons. These results indicate that the vagus nerve plays a crucial role in determining peripheral energy balance. The efficiency of ghrelin and CCK signal transduction may depend on the balance of their respective plasma concentration and/or on interactions between GHS-R and CCK-AR. PMID- 15890777 TI - CD147 is a regulatory subunit of the gamma-secretase complex in Alzheimer's disease amyloid beta-peptide production. AB - gamma-Secretase is a membrane protein complex that cleaves the beta-amyloid precursor protein (APP) within the transmembrane region, after prior processing by beta-secretase, producing amyloid beta-peptides Abeta(40) and Abeta(42). Errant production of Abeta-peptides that substantially increases Abeta(42) production has been associated with the formation of amyloid plaques in Alzheimer's disease patients. Biophysical and genetic studies indicate that presenilin-1, which contains the proteolytic active site, and three other membrane proteins [nicastrin, anterior pharynx defective-1 (APH-1), and presenilin enhancer-2 (PEN-2)] are required to form the core of the active gamma secretase complex. Here, we report the purification of the native gamma-secretase complexes from HeLa cell membranes and the identification of an additional gamma secretase complex subunit, CD147, a transmembrane glycoprotein with two Ig-like domains. The presence of this subunit as an integral part of the complex itself was confirmed through coimmunoprecipitation studies of the purified protein from HeLa cells and of solubilized complexes from other cell lines such as neural cell HCN-1A and HEK293. Depletion of CD147 by RNA interference was found to increase the production of Abeta peptides without changing the expression level of the other gamma-secretase components or APP substrates whereas CD147 overexpression had no statistically significant effect on Abeta-peptide production, other gamma secretase components or APP substrates, indicating that the presence of the CD147 subunit within the gamma-secretase complex down-modulates the production of Abeta peptides. PMID- 15890778 TI - The importance of weak absorption features in promoting tropospheric radical production. AB - Atmospheric field measurement and modeling studies have long noted discrepancies between observation and predictions of OH and HO(2) concentrations in the atmosphere. Novel photochemical mechanisms have been proposed to explain these differences. Although inclusion of these additional sources improves agreement, they are unable to fully account for the observations. We report and demonstrate the importance of weak electronic absorption features, normally ignored or not measured, in contributing to significant OH radical production. Experiments on methyl hydroperoxide, a prototypical organic peroxide in large abundance in the troposphere, highlights how photochemistry in the neglected electronic absorption tail makes an important addition to the tropospheric OH budget. The present results underscore the need to measure absorption cross sections for atmospheric molecules over a wider dynamic range, especially over the wavelength regions where the solar flux is high, to fully quantitate their contributions to atmospheric photochemistry. PMID- 15890779 TI - Serum protein markers for early detection of ovarian cancer. AB - Early diagnosis of epithelial ovarian cancer (EOC) would significantly decrease the morbidity and mortality from this disease but is difficult in the absence of physical symptoms. Here, we report a blood test, based on the simultaneous quantization of four analytes (leptin, prolactin, osteopontin, and insulin-like growth factor-II), that can discriminate between disease-free and EOC patients, including patients diagnosed with stage I and II disease, with high efficiency (95%). Microarray analysis was used initially to determine the levels of 169 proteins in serum from 28 healthy women, 18 women newly diagnosed with EOC, and 40 women with recurrent disease. Evaluation of proteins that showed significant differences in expression between controls and cancer patients by ELISA assays yielded the four analytes. These four proteins then were evaluated in a blind cross-validation study by using an additional 106 healthy females and 100 patients with EOC (24 stage I/II and 76 stage III/IV). Upon sample decoding, the results were analyzed by using three different classification algorithms and a binary code methodology. The four-analyte test was further validated in a blind binary code study by using 40 additional serum samples from normal and EOC cancer patients. No single protein could completely distinguish the cancer group from the healthy controls. However, the combination of the four analytes exhibited the following: sensitivity 95%, positive predictive value (PPV) 95%, specificity 95%, and negative predictive value (NPV) 94%, a considerable improvement on current methodology. PMID- 15890780 TI - Human modification of global water vapor flows from the land surface. AB - It is well documented that human modification of the hydrological cycle has profoundly affected the flow of liquid water across the Earth's land surface. Alteration of water vapor flows through land-use changes has received comparatively less attention, despite compelling evidence that such alteration can influence the functioning of the Earth System. We show that deforestation is as large a driving force as irrigation in terms of changes in the hydrological cycle. Deforestation has decreased global vapor flows from land by 4% (3,000 km(3)/yr), a decrease that is quantitatively as large as the increased vapor flow caused by irrigation (2,600 km(3)/yr). Although the net change in global vapor flows is close to zero, the spatial distributions of deforestation and irrigation are different, leading to major regional transformations of vapor-flow patterns. We analyze these changes in the light of future land-use-change projections that suggest widespread deforestation in sub-Saharan Africa and intensification of agricultural production in the Asian monsoon region. Furthermore, significant modification of vapor flows in the lands around the Indian Ocean basin will increase the risk for changes in the behavior of the Asian monsoon system. This analysis suggests that the need to increase food production in one region may affect the capability to increase food production in another. At the scale of the Earth as a whole, our results emphasize the need for climate models to take land use change, in both land cover and irrigation, into account. PMID- 15890781 TI - The frequencies of calcium oscillations are optimized for efficient calcium mediated activation of Ras and the ERK/MAPK cascade. AB - Ras proteins are binary switches that, by cycling through inactive GDP- and active GTP-bound conformations, regulate multiple cellular signaling pathways, including those that control growth and differentiation. For some time, it has been known that receptor-mediated increases in the concentration of intracellular free calcium ([Ca(2+)](i)) can modulate Ras activation. Increases in [Ca(2+)](i) often occur as repetitive Ca(2+) spikes or oscillations. Induced by electrical or receptor stimuli, these repetitive Ca(2+) oscillations increase in frequency with the amplitude of receptor stimuli, a phenomenon critical for the induction of selective cellular functions. Here, we show that Ca(2+) oscillations are optimized for Ca(2+)-mediated activation of Ras and signaling through the extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) cascade. We present additional evidence that Ca(2+) oscillations reduce the effective Ca(2+) threshold for the activation of Ras and that the oscillatory frequency is optimized for activation of Ras and the ERK/MAPK pathway. Our results describe a hitherto unrecognized link between complex Ca(2+) signals and the modulation of the Ras/ERK/MAPK signaling cascade. PMID- 15890782 TI - An N-ethyl-N-nitrosourea screen for genes involved in variegation in the mouse. AB - We have developed a sensitized screen to identify genes involved in gene silencing, using random N-ethyl-N-nitrosourea mutagenesis on mice carrying a variegating GFP transgene. The dominant screen has produced six mutant lines, including both suppressors and enhancers of variegation. All are semidominant and five of the six are homozygous embryonic lethal. In one case, the homozygous lethality depends on sex: homozygous females die at midgestation and display abnormal DNA methylation of the X chromosome, whereas homozygous males are viable. Linkage analysis reveals that the mutations map to unique chromosomal locations. We have studied the effect of five of the mutations on expression of an endogenous allele known to be sensitive to epigenetic state, agouti viable yellow. In all cases, there is an effect on penetrance, and in most cases, parent of origin and sex-specific effects are detected. This screen has identified genes that are involved in epigenetic reprogramming of the genome, and the behavior of the mutant lines suggests a common mechanism between X inactivation and transgene and retrotransposon silencing. Our findings raise the possibility that the presence or absence of the X chromosome in mammals affects the establishment of the epigenetic state at autosomal loci by acting as a sink for proteins involved in gene silencing. The study demonstrates the power of sensitized screens in the mouse not only for the discovery of novel genes involved in a particular process but also for the elucidation of the biology of that process. PMID- 15890783 TI - Liquid-crystal imaging of molecular-tilt ordering in self-assembled lipid tubules. AB - Self-assembled cylindrical tubules of chiral phospholipids are interesting supramolecular structures. Understanding the molecular-tilt order is a key step in controlling the size and shape of the tubules and designing new functional materials. The current theories based on the chiral interactions, coupled with molecular tilt, have predicted that the tubules could have both uniform and modulated tilt states. Here, we image the molecular-tilt order in the self assembled tubules of a chiral phospholipid by using liquid crystals as an optical amplification probe. We demonstrate that the organization of the molecular-tilt azimuth in the lipid tubules can induce an azimuthal orientation in the liquid crystals. Both uniform and modulated tilt states of the lipid tubules are observed after liquid-crystal optical amplification. PMID- 15890784 TI - Resolvin E1, an endogenous lipid mediator derived from omega-3 eicosapentaenoic acid, protects against 2,4,6-trinitrobenzene sulfonic acid-induced colitis. AB - Resolvin E1 (RvE1; 5S,12R,18R-trihydroxyeicosapentaenoic acid) is an antiinflammatory lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid (EPA). At the local site of inflammation, aspirin treatment enhances EPA conversion to 18R-oxygenated products, including RvE1, which carry potent antiinflammatory signals. Here, we obtained evidence for reduced leukocyte infiltration in a mouse peritonitis model, where the administration of EPA and aspirin initiated the generation of RvE1 in the exudates. Similar results were obtained with the administration of synthetic RvE1, which blocked leukocyte infiltration. RvE1 also protected against the development of 2,4,6 trinitrobenzene sulfonic acid-induced colitis. The beneficial effect was reflected by increased survival rates, sustained body weight, improvement of histologic scores, reduced serum anti-2,4,6-trinitrobenzene sulfonic acid IgG, decreased leukocyte infiltration, and proinflammatory gene expression, including IL-12 p40, TNF-alpha, and inducible nitric oxide synthase. Thus, the endogenous lipid mediator RvE1 counter-regulates leukocyte-mediated tissue injury and proinflammatory gene expression. These findings show an endogenous mechanism that may underlie the beneficial actions of omega-3 EPA and provide targeted approaches for the treatment of intestinal inflammation. PMID- 15890785 TI - AZF deletions and Y chromosomal haplogroups: history and update based on sequence. AB - AZF deletions are genomic deletions in the euchromatic part of the long arm of the human Y chromosome (Yq11) associated with azoospermia or severe oligozoospermia. Consequently, it can be assumed that these deletions remove Y chromosomal genes required for spermatogenesis. However, these 'classical' or 'complete' AZF deletions, AZFa, AZFb and AZFc, represent only a subset of rearrangements in Yq11. With the benefit of the Y chromosome sequence, more rearrangements (deletions, duplications, inversions) inside and outside the classical AZF deletion intervals have been elucidated and intra-chromosomal non allelic homologous recombinations (NAHRs) of repetitive sequence blocks have been identified as their major cause. These include duplications in AZFa, AZFb and AZFc and the partial AZFb and AZFc deletions of which some were summarized under the pseudonym 'gr/gr' deletions. At least some of these rearrangements are associated with distinct Y chromosomal haplogroups and are present with similar frequencies in fertile and infertile men. This suggests a functional redundancy of the AZFb/AZFc multi-copy genes. Alternatively, the functional contribution(s) of these genes to human spermatogenesis might be different in men of different Y haplogroups. That raises the question whether, the frequency of Y haplogroups with different AZF gene contents in distinct human populations leads to a male fertility status that varies between populations or whether, the presence of the multiple Y haplogroups implies a balancing selection via genomic deletion/amplification mechanisms. PMID- 15890786 TI - ACTH, alpha-MSH, and control of cortisol release: cloning, sequencing, and functional expression of the melanocortin-2 and melanocortin-5 receptor in Cyprinus carpio. AB - Cortisol release from fish head kidney during the acute phase of the stress response is controlled by the adrenocorticotropic hormone (ACTH) from the pituitary pars distalis (PD). Alpha-melanocyte-stimulating hormone (alpha-MSH) and beta-endorphin, from the pars intermedia (PI), have been implicated in cortisol release during the chronic phase. The present study addresses the regulation of cortisol release by ACTH and alpha-MSH in common carp (Cyprinus carpio) and includes characterization of their receptors, namely, the melanocortin-2 and melanocortin-5 receptors (MC2R and MC5R). We could not demonstrate corticotropic activity of alpha-MSH, beta-endorphin, and combinations of these. We do show a corticotrope in the PI, but its identity is as yet uncertain. Carp restrained for 1 and 7 days showed elevated plasma cortisol and alpha-MSH levels; cortisol is still elevated but lower at day 7 than day 1 of restraint. Interrenal response capacity is unaffected, as estimated by stimulation with a maximum dose ACTH in a superfusion setup. MC2R and MC5R appear phylogenetically well conserved. MC2R is predominantly expressed in head kidney; a low abundance was found in spleen and kidney. MC5R is expressed in brain, pituitary PD, kidney, and skin. Quantitative PCR analysis of MC2R and MC5R expression in the head kidney of restrained fish reveals MC2R mRNA downregulation after 7 days restraint, in line with lower plasma cortisol levels seen. We discuss regulation of corticosteroid production from a phylogenetic perspective. We propose that increased levels of alpha-MSH exert a positive feedback on hypothalamic corticotropin-releasing hormone release to sustain a mild stress axis activity. PMID- 15890787 TI - Ionizing radiation alters myofilament calcium sensitivity in vascular smooth muscle: potential role of protein kinase C. AB - Radiation exposure increases vascular responsiveness, and this change involves endothelial damage, as well as direct effects on vascular smooth muscle. In this study, we tested the hypothesis that myofilament Ca(2+) sensitivity in vascular smooth muscle is increased from single whole body gamma irradiation (6 Gy). We measured contractile responses from intact and permeabilized rat thoracic aortic rings combined with cytosolic Ca(2+) ([Ca(2+)](i)) measurements. The sensitivity to KCl and phenylephrine increased significantly in tissues from animals on the 9th and 30th days postirradiation compared with control. Irradiation also significantly increased Ca(2+) sensitivity in beta-escin permeabilized smooth muscle on the 9th and 30th days postirradiation. Inhibitors of protein kinase C, chelerythrine, and staurosporine, had no effect on the pCa-tension curves in control permeabilized tissues but significantly decreased Ca(2+) sensitivity in permeabilized tissues on the 9th and 30th days postirradiation. Phorbol dibutyrate (PDBu, 10(-7) M) increased Ca(2+) sensitivity in control skinned smooth muscle but was without effect in irradiated vascular rings. Simultaneous measurement of contractile force and [Ca(2+)](i) showed that myofilament Ca(2+) sensitivity defined as the ratio of force change to [Ca(2+)](i) significantly increased following gamma-irradiation. PDBu (10(-6) M) stimulation of intact aorta produced a sustained contraction, while the increase in [Ca(2+)](i) was transient. In irradiated tissues, PDBu-induced contractions were greater than those seen in control tissues but there was no elevation in [Ca(2+)](i). Taken together, these data strongly support the hypothesis that irradiation increases the sensitivity of vascular smooth muscle myofilaments to Ca(2+) and this effect is dependent on activation of protein kinase C. PMID- 15890788 TI - Angiotensin II and nitric oxide in neural control of intrarenal blood flow. AB - We investigated the roles of the renin-angiotensin system and the significance of interactions between angiotensin II and nitric oxide, in responses of regional kidney perfusion to electrical renal nerve stimulation (RNS) in pentobarbital sodium-anesthetized rabbits. Under control conditions, RNS (0.5-8 Hz) reduced total renal blood flow (RBF; -89 +/- 3% at 8 Hz) and cortical perfusion (CBF; -90 +/- 2% at 8 Hz) more than medullary perfusion (MBF; -55 +/- 5% at 8 Hz). Angiotensin II type 1 (AT(1))-receptor antagonism (candesartan) blunted RNS induced reductions in RBF (P = 0.03), CBF (P = 0.007), and MBF (P = 0.04), particularly at 4 and 8 Hz. Nitric oxide synthase inhibition with N(G)-nitro-L arginine (L-NNA) enhanced RBF (P = 0.003), CBF (P = 0.001), and MBF (P = 0.03) responses to RNS, particularly at frequencies of 2 Hz and less. After candesartan pretreatment, L-NNA significantly enhanced RNS-induced reductions in RBF (P = 0.04) and CBF (P = 0.007) but not MBF (P = 0.66). Renal arterial infusion of angiotensin II (5 ng.kg(-1).min(-1)) selectively enhanced responses of MBF to RNS in L-NNA-pretreated but not in vehicle-pretreated rabbits. In contrast, greater doses of angiotensin II (5-15 ng.kg(-1).min(-1)) blunted responses of MBF to RNS in rabbits with intact nitric oxide synthase. These results suggest that endogenous angiotensin II enhances, whereas nitric oxide blunts, neurally mediated vasoconstriction in the renal cortical and medullary circulations. In the renal medulla, but not the cortex, angiotensin II also appears to be able to blunt neurally mediated vasoconstriction. PMID- 15890789 TI - Differentiation of C2C12 myoblasts is critically regulated by FAK signaling. AB - This study examined whether focal adhesion kinase (FAK) plays a role in the differentiation of C(2)C(12) myoblasts into myotubes. Differentiation of C(2)C(12) myoblasts induced by switch to differentiation culture medium was accompanied by a transient reduction of FAK phosphorylation at Tyr-397 (to approximately 50%, at 1 and 2 h), followed by an increase thereafter (to 240% up to 5 days), although FAK protein expression remained unchanged. FAK and phosphorylated FAK were found at the edge of lamellipodia in proliferating cells, whereas the later increase in FAK phosphorylation in differentiating cells was accompanied by its preferential location at the tip of well-organized actin stress fibers. Hyperexpression of FAK autophosphorylation site (Tyr-397) mutant (MT-FAK) reduced FAK phosphorylation at Tyr-397 in proliferating cells and was accompanied by reduction of cyclin D1 and increase of myogenin expression. These cells failed to progress to myotubes in differentiation medium. In contrast, hyperexpression of a wild-type FAK construction (WT-FAK) increased baseline and abolished the transient reduction of FAK phosphorylation at Tyr-397 in serum starved C(2)C(12) cells. Cells transfected with WT-FAK failed to reduce cyclin D1 and to increase myogenin expression, as well as to progress to terminal differentiation in differentiation medium. These data indicate that FAK signaling plays a critical role in the control of cell cycle as well as in the progression of C(2)C(12) cells to terminal differentiation. Transient inhibition of FAK phosphorylation at Tyr-397 contributes to trigger the myogenic genetic program, but its later activation is also central to terminal differentiation into myotubes. PMID- 15890790 TI - Genetic and physiological insights into the metabolic syndrome. AB - The metabolic syndrome (MetS) is a common phenotype that is clinically defined by threshold values applied to measures of central obesity, dysglycemia, dyslipidemia, and/or elevated blood pressure, which must be present concurrently in any one of a variety of combinations. Insulin resistance, although not a defining component of the MetS, is nonetheless considered to be a core feature. MetS is important because it is rapidly growing in prevalence and is strongly related to the development of cardiovascular disease. To define etiology, pathogenesis and expression of MetS, we have studied patients, specifically Canadian families and communities. One example is familial partial lipodystrophy (FPLD), a rare monogenic form of insulin resistance caused by mutations in either LMNA, encoding nuclear lamin A/C (subtype FPLD2), or in PPARG, encoding peroxisomal proliferator-activated receptor-gamma (subtype FPLD3). Because it evolves slowly and recapitulates key clinical and biochemical attributes, FPLD seems to be a useful monogenic model of MetS. A second example is the disparate MetS prevalence between two Canadian aboriginal groups that is mirrored by disparate prevalence of diabetes and cardiovascular disease. Careful phenotypic evaluation of such special cases of human MetS by using a wide range of diagnostic methods, an approach called "phenomics," may help uncover early presymptomatic disease biomarkers, which in turn might reveal new pathways and targets for interventions for MetS, diabetes, and atherosclerosis. PMID- 15890791 TI - Calcium/calmodulin-dependent kinase II mediates critical components of the hypoxic ventilatory response within the nucleus of the solitary tract in adult rats. AB - Calcium/calmodulin-dependent kinase II (CaMKII) is an ubiquitous second messenger that is highly expressed in neurons, where it has been implicated in some of the pathways regulating neuronal discharge as well as N-methyl-d-aspartate receptor mediated synaptic plasticity. The full expression of the mammalian hypoxic ventilatory response (HVR) requires intact central relays within the nucleus of the solitary tract (NTS), and neural transmission of hypoxic afferent input is mediated by glutamatergic receptor activity, primarily through N-methyl-D aspartate receptors. To examine the functional role of CaMKII in HVR, KN-93, a highly selective antagonist of CaMKII, was microinjected in the NTS via bilaterally placed osmotic pumps in freely behaving adult male Sprague-Dawley rats for 3 days. Vehicle-loaded osmotic pumps were surgically placed in control animals, and adequate placement of cannulas was ascertained for all animals. HVR was measured using whole body plethysmography during exposure to 10% O(2)-balance N(2) for 20 min. Compared with control rats, KN-93 administration elicited marked attenuations of peak HVR (pHVR) but did not modify normoxic minute ventilation. Differences in pHVR were primarily attributable to diminished respiratory frequency recruitments during pHVR without significant differences in tidal volume. These findings indicate that CaMKII activation in the NTS mediates respiratory frequency components of the ventilatory response to acute hypoxia; however, CaMKII activity does not appear to underlie components of normoxic ventilation. PMID- 15890792 TI - Temporal dynamics of late-night photic stimulation of the human circadian timing system. AB - The light-dark cycle is the primary synchronizing factor that keeps the internal circadian pacemaker appropriately aligned with the environmental 24-h day. Although it is known that ocular light exposure can effectively shift the human circadian pacemaker and do so in an intensity-dependent manner, the curve that describes the relationship between light intensity and pacemaker response has not been fully characterized for light exposure in the late biological night. We exposed subjects to 3 consecutive days of 5 h of experimental light, centered 1.5 h after the timing of the fitted minimum of core body temperature, and show that such light can phase advance shift the human circadian pacemaker in an intensity dependent manner, with a logistic model best describing the relationship between light intensity and phase shift. A similar sigmoidal relationship is also observed between light intensity and the suppression of plasma melatonin concentrations that occurs during the experimental light exposure. As with a simpler, 1-day light exposure during the early biological night, our data indicate that the human circadian pacemaker is highly sensitive even to typical room light intensities during the late biological night, with approximately 100 lux evoking half of the effects observed with light 10 times as bright. PMID- 15890793 TI - Short-term stress experienced at time of immunization induces a long-lasting increase in immunologic memory. AB - It would be extremely beneficial if one could harness natural, endogenous, health promoting defense mechanisms to fight disease and restore health. The psychophysiological stress response is the most underappreciated of nature's survival mechanisms. We show that acute stress experienced before primary immunization induces a long-lasting increase in immunity. Compared with controls, mice restrained for 2.5 h before primary immunization with keyhole limpet hemocyanin (KLH) show a significantly enhanced immune response when reexposed to KLH 9 mo later. This immunoenhancement is mediated by an increase in numbers of memory and effector helper T cells in sentinel lymph nodes at the time of primary immunization. Further analyses show that the early stress-induced increase in T cell memory may stimulate the robust increase in infiltrating lymphocyte and macrophage numbers observed months later at a novel site of antigen reexposure. Enhanced leukocyte infiltration may be driven by increased levels of the type 1 cytokines, IL-2 and IFN-gamma, and TNF-alpha, observed at the site of antigen reexposure in animals that had been stressed at the time of primary immunization. In contrast, no differences were observed in type 2 cytokines, IL-4 or IL-5. Given the importance of inducing long-lasting increases in immunologic memory during vaccination, we suggest that the neuroendocrine stress response is nature's adjuvant that could be psychologically and/or pharmacologically manipulated to safely increase vaccine efficacy. These studies introduce the novel concept that a psychophysiological stress response is nature's fundamental survival mechanism that could be therapeutically harnessed to augment immune function during vaccination, wound healing, or infection. PMID- 15890794 TI - Age-specific changes in the regulation of LH-dependent testosterone secretion: assessing responsiveness to varying endogenous gonadotropin output in normal men. AB - Pulsatile and thus total testosterone (Te) secretion declines in older men, albeit for unknown reasons. Analytical models forecast that aging may reduce the capability of endogenous luteinizing hormone (LH) pulses to stimulate Leydig cell steroidogenesis. This notion has been difficult to test experimentally. The present study used graded doses of a selective gonadotropin releasing hormone (GnRH)-receptor antagonist to yield four distinct strata of pulsatile LH release in each of 18 healthy men ages 23-72 yr. Deconvolution analysis was applied to frequently sampled LH and Te concentration time series to quantitate pulsatile Te secretion over a 16-h interval. Log-linear regression was used to relate pulsatile LH secretion to attendant pulsatile Te secretion (LH-Te drive) across the four stepwise interventions in each subject. Linear regression of the 18 individual estimates of LH-Te feedforward dose-response slopes on age disclosed a strongly negative relationship (r = -0.721, P < 0.001). Accordingly, the present data support the thesis that aging in healthy men attenuates amplitude-dependent LH drive of burst-like Te secretion. The experimental strategy of graded suppression of neuroglandular outflow may have utility in estimating dose response adaptations in other endocrine systems. PMID- 15890795 TI - Metabolism and thermoregulation during fasting in king penguins, Aptenodytes patagonicus, in air and water. AB - We measured oxygen consumption rate (Vo(2)) and body temperatures in 10 king penguins in air and water. Vo(2) was measured during rest and at submaximal and maximal exercise before (fed) and after (fasted) an average fasting duration of 14.4 +/- 2.3 days (mean +/- 1 SD, range 10-19 days) in air and water. Concurrently, we measured subcutaneous temperature and temperature of the upper (heart and liver), middle (stomach) and lower (intestine) abdomen. The mean body mass (M(b)) was 13.8 +/- 1.2 kg in fed and 11.0 +/- 0.6 kg in fasted birds. After fasting, resting Vo(2) was 93% higher in water than in air (air: 86.9 +/- 8.8 ml/min; water: 167.3 +/- 36.7 ml/min, P < 0.01), while there was no difference in resting Vo(2) between air and water in fed animals (air: 117.1 +/- 20.0 ml O(2)/min; water: 114.8 +/- 32.7 ml O(2)/min, P > 0.6). In air, Vo(2) decreased with M(b), while it increased with M(b) in water. Body temperature did not change with fasting in air, whereas in water, there were complex changes in the peripheral body temperatures. These latter changes may, therefore, be indicative of a loss in body insulation and of variations in peripheral perfusion. Four animals were given a single meal after fasting and the temperature changes were partly reversed 24 h after refeeding in all body regions except the subcutaneous, indicating a rapid reversal to a prefasting state where body heat loss is minimal. The data emphasize the importance in considering nutritional status when studying king penguins and that the fasting-related physiological changes diverge in air and water. PMID- 15890796 TI - Vitamin D receptor FokI genotype influences bone mineral density response to strength training, but not aerobic training. AB - To determine the influence of the vitamin D receptor (VDR) gene FokI and BsmI genotype on bone mineral density response to two exercise training modalities, 206 healthy men and women (50-81 years old) were studied before and after approximately 5-6 months of either aerobic exercise training (AT) or strength training (ST). A totla of 123 subjects completed AT (51 men, 72 women) and 83 subjects completed ST (40 men, 43 women). DNA was extracted from blood samples of all subjects and genotyping was performed at the VDR FokI and BsmI locus to determine its association to training response. Total body, greater trochanter and femoral neck bone mineral density (BMD) were measured before and after both training programmes using dual-energy X-ray absorptiometry. VDR BsmI genotype was not significantly related to BMD at baseline or after ST or AT. However, VDR FokI genotype was significantly related to ST- but not AT-induced changes in femoral neck BMD (P < 0.05). The heterozygotes (Ff) in the ST group approached a significantly greater increase in femoral neck BMD (P = 0.058) compared to f homozygotes. There were no significant genotype relationships in the AT group. These data indicate that VDR FokI genotype may influence femoral neck BMD response to ST, but not AT. PMID- 15890797 TI - Oxidative stress and vascular remodelling. AB - Oxidative stress plays an important role in the pathophysiology of vascular diseases. Reactive oxygen species, especially superoxide anion and hydrogen peroxide, are important signalling molecules in cardiovascular cells. Enhanced superoxide production increases nitric oxide inactivation and leads to an accumulation of peroxynitrites and hydrogen peroxide. Reactive oxygen species participate in growth, apoptosis and migration of vascular smooth muscle cells, in the modulation of endothelial function, including endothelium-dependent relaxation and expression of proinflammatory phenotype, and in the modification of the extracellular matrix. All these events play important roles in vascular diseases such as hypertension, suggesting that the sources of reactive oxygen species and the signalling pathways that they modify may represent important therapeutic targets. Potential sources of vascular superoxide production include NADPH-dependent oxidases, xanthine oxidases, lipoxygenases, mitochondrial oxidases and nitric oxide synthases. Studies performed during the last decade have shown that NADPH oxidase is the most important source of superoxide anion in phagocytic and vascular cells. Evidence from experimental animal and human studies suggests a significant role of NADPH oxidase activation in the vascular remodelling and endothelial dysfunction found in cardiovascular diseases. PMID- 15890798 TI - Intracellular mechanisms involved in vascular remodelling of resistance arteries in hypertension: role of angiotensin II. AB - Resistance arteries undergo structural changes (vascular remodelling) in hypertension. These changes involve media thickening, reduced lumen diameter and consequent increased media:lumen ratio. Cellular processes underlying these events include altered vascular smooth muscle cell (VSMC) growth, migration, differentiation and increased extracellular matrix abundance. Another factor contributing to remodelling is inflammation, associated with macrophage infiltration, fibrosis and increased expression of redox-sensitive pro inflammatory genes. Among the factors involved in arterial remodelling, angiotensin (Ang) II appears to be one of the most important. Ang II, a multifunctional peptide with pleiotropic actions, modulates vasomotor tone, cell growth, apoptosis/anoikis, cell migration and extracellular matrix deposition. It is pro-inflammatory and it stimulates production of growth factors and vasoactive agents. The multiple actions of Ang II are mediated via complex intracellular signalling pathways including stimulation of the phosholipase C (PLC)-inositol 1,4,5-trisphosphate (IP3)-1,2-diacylglycerol (DAG) cascade, mitogen-activated protein (MAP) kinases, tyrosine kinases and RhoA/Rho kinase. Furthermore, Ang II elicits many of its (patho)physiological effects by stimulating reactive oxygen species (*O2- and H2O2) generation through activation of vascular NAD(P)H oxidase. *O2- and H2O2 in turn influence downstream signalling molecules including transcription factors, tyrosine kinases/phosphatases, Ca2+ channels and MAP kinases. Interaction between these systems is complex and dysregulation at any level may contribute to vascular remodelling. Targeting such molecules/pathways could prevent or induce regression of hypertensive vascular damage thereby ameliorating development of hypertension and preventing target organ damage. The present review discusses the role of Ang II in remodelling of resistance arteries, focusing on some signalling pathways involved in vascular growth and inflammation in hypertension. PMID- 15890799 TI - Influence of elastin on rat small artery mechanical properties. AB - We have previously developed a method for estimating elastin content and organization in resistance arteries, where it is a minor component. The aim of the present study was to validate the method against a quantitative assay and to determine the relative importance of elastin content and organization for intrinsic elasticity of small arteries. Mesenteric third order branches (from 10 day-old, 1- and 6-month-old rats) and middle cerebral arteries (from 6-month-old rats) were pressurized. beta-Values were calculated from stress-strain relationships and used as indicators of intrinsic stiffness. The same pressure fixed arteries were used to estimate elastin content and organization in the internal elastic lamina with confocal microscopy. Collagen and elastin contents were determined by Picrosirius Red staining and radioimmunoassay for desmosine, respectively. Confocal and desmosine assays gave similar results: no difference in elastin content of mesenteric vessels from 1- and 6-month-old rats, and a significant reduction in cerebral compared to mesenteric arteries. For all parameters (elastin and collagen content, fenestrae area and internal elastic lamina thickness) the best correlation was found between beta-values and fenestrae size. These data suggest that in small arteries: (1) confocal microscopy can be used as a method for the simultaneous study of changes in elastin content and organization; and (2) elastin organization might be a key determinant of intrinsic elastic properties. PMID- 15890800 TI - Joint analysis of two breed cross populations in pigs to improve detection and characterization of quantitative trait loci. AB - The purpose of this study was to develop and implement least squares interval mapping models for joint analysis of breed cross QTL mapping populations and to evaluate the effect of joint analysis on QTL detected for economic traits in data from two breed crosses in pigs. Data on 26 growth, carcass composition, and meat quality traits from F2 crosses between commercially relevant pig breeds were used: a Berkshire x Yorkshire cross at Iowa State University (ISU) and a Berkshire x Duroc cross at the University of Illinois (UOI). All animals were genotyped for a total of 39 (ISU) and 32 (UOI) markers on chromosomes 2, 6, 13, and 18. Marker linkage maps derived from the individual and joint data were similar with regard to order and relative position, but some differences in absolute distances existed. Maps from the joint data were used in all analyses. The individual and joint data sets were analyzed using several least squares interval-mapping models: line-cross (LC) models with Mendelian and parent-of origin effects; halfsib models (HS); and combined models (CB) that included LC and HS effects. Lack-of-fit tests between the models were used to characterize QTL for mode of expression and to identify segregation of QTL within parental breeds. A total of 26 (8), 47 (18), and 53 (16) QTL were detected at the 5% chromosome (genome)-wise level in the ISU, UOI, and joint data for the 26 analyzed traits. Of the 53 QTL detected in the joint data, only six were detected in both populations and for many, allele effects differed between the two crosses. Despite the lack of overlap between the two populations, joint analysis resulted in an increase in significance for many QTL, including detection of ten QTL that did not reach significance in either population. Confidence intervals for position also were smaller for several QTL. In contrast, 24 QTL, most of which were detected at chromosome-wise levels in the ISU or UOI population, were not detected in the joint data. Presence of paternally expressed QTL near the IGF2 region of SSC2 was confirmed, with major effects on backfat and loin muscle area, particularly in the UOI population, as well as one or more QTL for carcass composition in the distal arm of Chromosome 6. Results of this study suggest that joint analysis using a range of QTL models increases the power of QTL mapping and QTL characterization, which helps to identify genes for subsequent marker assisted selection. PMID- 15890801 TI - Estimation of variance components including competitive effects of Large White growing gilts. AB - Records of on-test ADG of Large White gilts were analyzed to estimate variance components of direct and associative genetic effects. Models included the effects of contemporary group (farm-barn-batch), birth litter, pen group, and direct and associative additive genetic effects. The area of each pen was 14 m2. The additive genetic variance was a function of the number of competitors in a group, the additive relationships between the animal performing the record and its pen mates, and the additive relationships between pen mates. To partially account for differences in the number of pen mates, a covariable (qi = 1, 1/n, or 1/n(1/2)) was added to the associative genetic effect. There were 4,946 records from 2,409 litters and 362 pen groups. Pen group size ranged from 12 to 16 gilts. Analyses by REML converged very slowly. A grid search showed that the likelihood function was almost flat when the additive genetic associative effect was fitted. Estimates of direct and associative heritability were 0.15 and 0.03, respectively. Within the BLUPF90 family of programs, the mixed-model equations can be set up directly. For variance component estimation, simple programs (REMLF90 and GIBBSF90) worked without modifications, but more optimized programs did not. Estimates obtained using the three values of qi were similar. With the data structure available for this study and under an environment with relative low competition among animals, accurate estimation of associative genetic effects was not possible. Estimation of competitive effects with large pen size is difficult. The magnitude of competition effects may be larger in commercial populations, where housing is denser and food is limited. PMID- 15890802 TI - Influence of breed, heterozygosity, and disease incidence on estimates of variance components of respiratory disease in preweaned beef calves. AB - The objective of this study was to characterize genetic and environmental factors influencing bovine respiratory disease (BRD) in beef cattle. Records from nine purebred and three composite breeds and a variety of F1 and three-way crosses, including the progeny of 12 additional different sire breeds produced over a 20 yr period (1983 to 2002), were evaluated for breed and heterozygosity effects on the observed incidence of BRD. Heterozygosity fractions for calves and dams were defined by generalized breed origins: British, Continental, and tropically adapted. Variance components were estimated for each pure and composite breed, and across all breeds and crossbreeds. The effect of incidence of observed BRD was determined by comparing groups of low and high years of incidence. Respiratory disease in this herd followed a standard epidemiological pattern of initial introduction, reaching an epidemic stage at 70 to 170 d of age, followed by a period of rapid decrease to weaning. Estimates of heritability of incidence of BRD were low, ranging from 0.00 to 0.26, with overall estimates of 0.07 and 0.19 depending on the data set analyzed. The highest incidence of BRD in preweaned calves occurred in the Braunvieh breed (18.8%). The genetic correlation between the direct and maternal genetic effects was generally large and negative, suggesting dams genetically superior for resisting BRD raise calves that are more susceptible. Perhaps maternally superior dams provide passive immunity to their calves, which delays the development of the calves' direct immune system, making them more prone to BRD during the preweaning period. Heterozygosity of calves decreased the incidence of BRD compared with purebred cattle. Calves that were Continental x British or tropically adapted x British breeds had a lower incidence of BRD than did calves of British x British breeds. As the annual incidence of BRD increased, there was an associated increase in the heritability estimate. The estimated heritability based on an underlying continuous scale was large (h2 = 0.48), inferring response to selection for BRD resistance could be large if the phenotype for BRD resistance was known. PMID- 15890803 TI - Technical note: a dynamic model to predict the composition of fat-free matter gains in cattle. AB - Composition of empty BW (EBW) was described in terms of ether-extractable lipid (FAT) and fat-free matter (FFM), and the terms dEBW, dFAT, and dFFM were used to represent daily gains in these components. The dFFM comprised protein, water, and ash, and a model was developed to predict the composition of dFFM. The conceptual approach used in model development was based on experimental data that showed as cattle grew from birth to maturity: 1) the water content of FFM decreased and the protein and ash content increased; 2) the protein content of FFM increased at a decreasing rate; and 3) the protein-to-ash ratio in the fat-free DM was approximately constant. These results suggest that the protein content of dFFM would be high at birth and decrease at a decreasing rate as the animal grows. The protein content of dFFM was predicted as a function of the fraction of dEBW that was dFFM, FAT content of EBW, and dFFM. A fixed protein-to-ash ratio of 4.1:1 was used to calculate the quantity of ash, and water was obtained as a residual. Growth and body composition of Hereford x Angus steers from birth to 500 kg BW were simulated with a previously published model using the experimental growth data as input, and the model under discussion was used to predict the composition of dFFM. Predicted response curves of the EBW components over the growth period were similar in shape to observed data. Predicted curvilinearity in response of protein weight against FFM weight for Hereford x Angus steers was similar to observed data. The standard error about the regression of predicted on observed protein weight was 1.87 kg, and the average bias of the model was to underpredict protein weight by 0.64%. Compared with using a constant value for the protein fraction of dFFM, the model provided more accurate predictions of dEBW in an independent evaluation data set. PMID- 15890804 TI - Effects of feeding a blend of grains naturally contaminated with Fusarium mycotoxins on feed intake, metabolism, and indices of athletic performance of exercised horses. AB - An experiment was conducted to determine the effect of feeding blends of grains naturally contaminated with Fusarium mycotoxins to mature, exercised horses, and to test the efficacy of a polymeric glucomannan mycotoxin adsorbent (GM polymer) in preventing Fusarium mycotoxicoses. Six mature, mixed-breed mares with an average BW of 530 kg were assigned to one of three dietary treatments for 21 d in a replicated 3 x 3 Latin square design. Feed consumed each day was a combination of up to 3.5 kg of concentrates and 5.0 kg of mixed timothy/alfalfa hay (as-fed basis). The concentrates fed included 1) manage; 2) blend of contaminated grains; and 3) contaminated grains + 0.2% GM polymer (MTB-100, Alltech Inc., Nicholasville, KY). Concentrates containing contaminated grains averaged 11.0 ppm deoxynivalenol, 0.7 ppm 15-acetyldeoxynivalenol, and 0.8 ppm zearalenone (as-fed basis). Feed intake and BW were monitored over a 21-d period. Horses were maintained on a fixed exercise schedule throughout the experiment. At the end of the experiment, each horse completed a time-to-fatigue treadmill step test. Variables measured during pretest, each step of the test, and 5 and 10 min posttest were as follows: 1) time-to-fatigue, 2) heart rate, 3) hematological variables, and 4) serum lactate concentration. Each step consisted of 2 min of fast trot with a 2% increase in incline after each 2 min. Feed intake by horses fed contaminated grains was decreased compared with controls throughout the experiment (P < 0.05). Supplementation of 0.2% GM polymer to the contaminated diet did not alter feed intake by horses compared with those fed the unsupplemented contaminated diet. All hay was consumed regardless of concentrate fed. Weight loss from 0 to 21 d was observed in horses fed contaminated grains compared with controls (P < 0.05). No effect of diet was seen on variables used to measure athletic ability, although the results showed an expected response to exercise for a fit horse. We conclude that exercised horses are susceptible to Fusarium mycotoxicoses as indicated by appetite suppression and weight loss. PMID- 15890805 TI - Influence of supplementation of all-rac-alpha-tocopheryl acetate preweaning and vitamin C postweaning on alpha-tocopherol and immune responses of piglets. AB - This study was designed to test whether dietary maternal supplementation of all rac-alpha-tocopheryl acetate during lactation and dietary vitamin C supplementation after weaning could increase the alpha-tocopherol status pre- and postweaning and the immune responses of piglets postweaning. The experiment involved 12 crossbred sows that were fed increasing levels of all-rac-alpha tocopheryl (70, 150, and 250 IU/kg, as-fed basis) during lactation. After weaning (d 28 of age), litters were divided into two groups that were supplemented with or without vitamin C (500 mg/kg of feed, as-fed basis). Milk and blood samples were obtained from the sows during lactation. Pigs were bled at 4, 16, 28, 35, 42, and 49 d of age. Liver, heart, muscle, and s.c. adipose tissues were collected (on 28, 35, 42, and 49 d of age) and analyzed for alpha-tocopherol. On the same days, alveolar macrophages of the lungs were collected, and analyzed for the concentration of alpha-tocopherol and its stereoisomer composition, fatty acid composition, and release of prostaglandin E2, leukotriene B4, and thromboxane B2. Increasing dietary all-rac-alpha-tocopheryl acetate concentration increased the concentration of alpha-tocopherol in plasma (P = 0.02) and milk (P = 0.007) of sows, and the sow milk concentrations of alpha-tocopherol and vitamin A were greater on d 2 of lactation than later on during lactation. The plasma concentration of alpha-tocopherol in piglets decreased from d 4 to later on during suckling (P < 0.001) and again as the postweaning period progressed (P < 0.001). When lipid-standardized, plasma alpha-tocopherol was increased in piglets of sows fed 250 IU of all-rac-alpha-tocopheryl acetate compared with other sow groups (P = 0.005). At 28 d of age, alpha-tocopherol concentrations in tissues were increased with supplementation of the high dietary all-rac-alpha-tocopheryl acetate levels to the sows; however, after weaning, a decrease in alpha tocopherol concentration in most tissues (except liver) was observed, but the decrease tended to be less in the muscle (P = 0.099) and adipose tissue (P = 0.11) of piglets suckling sows fed 150 and 250 IU of all-rac-alpha-tocopheryl acetate. Vitamin C supplementation after weaning increased liver alpha-tocopherol (P = 0.01) and serum immunoglobulin M concentration (P = 0.04), and vitamin C supplementation increased the proportion of the RRR-alpha-tocopherol (P = 0.03) at the expense of the RRS-stereoisomer form (P = 0.05) of alpha-tocopherol in alveolar macrophages of the piglets. In conclusion, this study on maternal all rac-alpha-tocopheryl acetate and postweaning vitamin C supplementation suggests a nutritional strategy for increasing alpha-tocopherol status and immune responses of weaned piglets. PMID- 15890806 TI - Feed physical form and formic acid addition to the feed affect the gastrointestinal ecology and growth performance of growing pigs. AB - The effect of feeding a coarsely ground meal (COARSE) and a finely ground pelleted diet with 1.8% (as-fed basis) added formic acid (ACID) was compared with feeding a standard finely ground pelleted diet (STD) on the gastrointestinal ecology of growing pigs at different intervals after feeding. One hundred five castrated male growing-finishing pigs (initial BW 27 kg) were used. At a BW of 63 kg, 60 pigs were killed 0.5, 2.5, 4.5, 6.5, and 8.5 h after feeding, and samples from the gastrointestinal tract (GIT) were obtained. The remaining 45 pigs were kept on the experimental diets to a BW of 99 kg. Feeding the three diets resulted in a similar pattern of gastric pH with time, (i.e., highest pH values 0.5 h after feeding and decreasing values at the following sampling times, to reach a value of 2.12 at 8.5 h after feeding). The pH of the gastric digesta of pigs fed the ACID diet was below 4 at all sampling times, whereas the digesta from the other two dietary groups had values above pH 4 at the first sampling times. Feeding the ACID diet decreased the counts of total anaerobes in the proximal GIT (P < or = 0.007), and of lactic acid bacteria (P < or = 0.001), enterobacteria (P < or = 0.02), and yeasts (P < or = 0.01) along the GIT compared with feeding the other two diets. Feeding the COARSE diet stimulated the growth of total anaerobes and lactic acid bacteria in the stomach and distal small intestine increased the microbial diversity mainly in the stomach (P = 0.001), compared with feeding the other two diets (P < or = 0.09), and decreased the number of enterobacteria in the cecum compared with the STD diet (P = 0.03), with the same tendency in the mid-colon (P = 0.07). The concentration of lactic acid in the stomach was highest in the pigs fed the COARSE diet compared with the other two groups (P < 0.05). The concentration of formic acid was highest in the stomach and all segments of the small intestine of the pigs fed the ACID diet compared with those fed the STD and COARSE diets (P < 0.05). The results from this study suggest that feeding a coarsely ground diet and a finely ground diet with added formic acid affect the gastrointestinal ecology of pigs mainly by changing the environment in the proximal GIT. The presence of organic acids in the proximal GIT is a crucial factor contributing to the decrease in the number of enterobacteria along the GIT. The time after feeding at which samples are taken to measure characteristics describing the gastrointestinal ecology affects the results from the stomach and small intestine. PMID- 15890807 TI - The tryptophan requirement of growing and finishing barrows. AB - Five experiments were conducted to determine the true ileal digestible Trp (tidTrp) requirement of growing and finishing pigs fed diets (as-fed basis) containing 0.87% (Exp. 3), 0.70% (Exp. 4), 0.61% (Exp. 5), and 0.52% (Exp. 1 and 2) tidLys during the early-grower, late-grower, early-finisher, and late-finisher periods, respectively. Treatments were replicated with three or four replications, with three or four pigs per replicate pen. Treatment differences were considered significant at P = 0.10. Experiment 1 was conducted with 27 pigs (initial and final BW of 78.3 +/- 0.5 and 109.8 +/- 1.9 kg) to validate whether a corn-feather meal (FM) tidTrp-deficient (0.07%) diet, when supplemented with 0.07% crystalline l-Trp, would result in growth performance and carcass traits similar to a conventional corn-soybean meal (C-SBM) diet. Pigs fed the corn-FM diet without Trp supplementation had decreased growth performance and carcass traits, and increased plasma urea N (PUN) concentration. Supplementing the corn FM diet with Trp resulted in greater ADG and G:F than pigs fed the positive control C-SBM diet. Pigs fed the corn-FM diet had similar carcass traits as pigs fed the C-SBM diet, but loin muscle area was decreased and fat thickness was increased. In Exp. 2, 60 pigs (initial and final BW of 74.6 +/- 0.50 and 104.5 +/ 1.64 kg) were used to estimate the tidTrp requirement of finishing pigs. The levels of tidTrp used in Exp. 2 were 0.06, 0.08, 0.10, 0.12, or 0.14% (as-fed basis). Response variables were growth performance, PUN concentrations, and carcass traits and quality. For Exp. 2, the average of the estimates calculated by broken-line regression was 0.104% tidTrp. In Exp. 3, 4, and 5, barrows (n = 60, 60, or 80, respectively) were allotted to five dietary treatments supplemented with crystalline l-Trp at increments of 0.02%. The basal diets contained 0.13, 0.09, and 0.07% tidTrp (as-fed basis) in Exp. 3, 4, and 5, and initial BW of the pigs in these experiments were 30.9 +/- 0.7, 51.3 +/- 1.1, and 69.4 +/- 3.0 kg, respectively. The response variable was PUN, and the basal diet used in Exp. 3 and 4 contained corn, SBM, and Canadian field peas. The tidTrp requirements were estimated to be 0.167% for pigs weighing 30.9 kg, 0.134% for pigs weighing 51.3 kg, and 0.096% for pigs weighing 69.4 kg. Based on our data and a summary of the cited literature, we suggest the following total Trp and tidTrp requirement estimates (as-fed basis): 30-kg pigs, 0.21 and 0.18%; 50-kg pigs, 0.17 and 0.14%; 70-kg pigs, 0.13 and 0.11%; and in 90-kg pigs, 0.13 and 0.11%. PMID- 15890808 TI - Effect of diets containing linoleic acid- or oleic acid-rich oils on ruminal fermentation and nutrient digestibility, and performance and fatty acid composition of adipose and muscle tissues of finishing cattle. AB - Two trials were conducted to determine the effect of linoleic acid- or oleic acid rich safflower oil on ruminal fermentation, nutrient digestion, feedlot performance, carcass characteristics, and fatty acid composition of adipose and muscle tissues of beef cattle. In both trials, cattle were fed a finishing diet based on barley grain, wheat silage, and alfalfa hay. Oils were fed at 5% of dietary DM. In a metabolism trial, four ruminally and duodenally cannulated Angus crossbred steers were subjected to linoleic acid-rich oil or oleic acid-rich oil in a crossover design with covariate periods (no oil supplementation). In a finishing trial, 16 individually fed Angus crossbred steers and heifers (eight per diet) received linoleic acid- or oleic acid-rich oils during the last 86 d of a 116-d feeding period. Ruminal pH, ammonia concentration, protozoal counts, major VFA concentrations, acetate-to-propionate ratio, polysaccharide-degrading activities, microbial N flow to the duodenum, and the efficiency of microbial N synthesis in the rumen were not affected (P = 0.18 to 0.96) by type of oil. Type of oil had no effect on total-tract apparent digestion of nutrients (P = 0.46 to 0.98). Ruminal true nutrient digestibilities did not differ between oils (P = 0.15 to 0.99), except that the linoleic acid-rich oil decreased (P = 0.05) NDF digestibility. Dry matter intake, ADG, G:F, and carcass characteristics did not differ (P = 0.11 to 0.84) between the two oils. Overall, the difference in dietary fatty acids provided to the cattle produced few changes in tissue fatty acids. Weight percentages of c9t11 CLA were unaltered by the addition of linoleic acid to the diet compared with oleic acid, probably as a result of low vaccenic acid production in the rumen, as the pathway of biohydrogenation was apparently primarily through the t10 pathway. PMID- 15890809 TI - Wheat pasture bloat dynamics, in vitro ruminal gas production, and potential bloat mitigation with condensed tannins. AB - The aim of this study was to determine the effect of winter wheat (Triticum aestivum L.) forage growth stage, forage allowance, time of day, and commercial condensed tannins (CT) on steer bloat dynamics and in vitro ruminal gas production. Twenty-six crossbreed steers (Angus x Hereford x Salers; average initial BW = 194 +/- 26 kg) were used. Wheat forage allowances were either 18 kg (high forage allowance) and 6 kg (low forage allowance) of DM/(100 kg BW.d). In each bloat observation period, fresh wheat forage samples were hand-clipped to ground level in all study pastures for nutrient and in vitro ruminal gas production analyses. In vitro ruminal gas accumulation was measured at 0, 1, 2, 3, 4, 5, 6, and 12 h. Commercial CT was added at 0, 10, 15, and 20 mg of CT/g of DM. Bloat was scored once per week on two consecutive days at 0800 and 1500 during the vegetative stage and once every 2 wk during the reproductive stage of wheat development. Mean bloat score was calculated for each steer by time of day, stage of plant growth, and forage allowance. Bloat was detected in 65.8% of the observation periods. Average bloat scores were four and 2.5 times greater (P < 0.05) in cattle grazing at a high forage allowance than at a low forage allowance in the vegetative and reproductive growth phases of wheat, respectively. Rate of gas production was greater (P < 0.001) in the vegetative stage than in the reproductive stage. Steer bloat score was positively correlated with forage CP (r = 0.22; P < 0.05) and IVDMD (r = 0.32; P < 0.05). Rate of ruminal gas production was positively correlated (P < 0.01) to forage CP (r = 0.48), NPN (r = 0.40), soluble protein (r = 0.32), and IVDMD (r = 0.47). Conversely, negative correlations were found for forage DM (r = -0.20; P < 0.05), insoluble protein (r = -0.40), NDF (r = -0.69), and forage height (r = -0.49; P < 0.01) on the rate of ruminal gas production. Addition of CT at levels greater than 10 mg of CT/g of DM decreased (P < 0.05) the rate of in vitro ruminal gas and methane gas production after 5 h of incubation. Wheat pasture bloat is a complex disorder that varies across an array of forage and environmental conditions. Condensed tannins have the potential to decrease bloat by altering ruminal gas production and soluble protein digestibility from wheat forage. PMID- 15890810 TI - Relationship between fecal crude protein concentration and diet organic matter digestibility in cattle. AB - The positive relationship between fecal CP concentration and diet OM digestibility in cattle, which is based on increasing undigested microbial CP and decreasing fecal OM as OM digestibility increases, may be used as an indirect method for estimating diet OM digestibility from fecal CP concentration. Results of digestibility trials (445 individual observations) conducted at Hohenheim and Braunschweig, Germany, and at Gumpenstein, Austria, were used to study the relationship between CP concentration in feces (x, g/kg OM) and OM digestibility (y, %). The best fit was obtained with the curvilinear relationship y = ai 107.7e(-0.01515 x x), with a1 = 79.76 and a2 = 72.86 (R2 = 0.82; residual SD = 2.7; SE = 0.13), which takes into account the effects of location (i = 1 for Braunschweig and Hohenheim, and i = 2 for Gumpenstein). Dietary CP and crude fat concentration, and DMI had no effect on fecal CP content, whereas crude fiber content, proportion of concentrate in the diet, and forage type significantly affected CP concentration in feces; however, the magnitude of these effects was less than 2 percentage units, and the direction of the effect of proportion of concentrate in the diet was not uniform. The curvilinear relationship between fecal CP concentration (observed range, 100 to 300 g/kg of OM) and diet OM digestibility (observed range = 57 to 80%) may be used to estimate diet OM digestibility, particularly for field trials, as it requires no feed samples and does not physically restrict the animal. PMID- 15890811 TI - Characteristics of lambs fed concentrates or grazed on ryegrass to traditional or heavy slaughter weights. II. Wholesale cuts and tissue accretion. AB - Targhee x Hampshire lambs (average BW 24 +/- 1 kg) were used to determine the effect of finishing on concentrate or by grazing ryegrass forage on slaughter weights of 52 kg (N) or 77 kg (H) on tissue accretion and lamb wholesale cutout. When fed to similar slaughter weights, the wholesale cuts of concentrate-fed lambs were heavier (P < 0.05) than the same cuts from forage-fed lambs; however, when expressed as a percentage of side weight, carcasses of forage-fed lambs had a higher (P < 0.001) percentage of leg than concentrate-fed lambs. Increasing slaughter weight from 52 to 77 kg resulted in a 1-kg increase in loin weight for lambs finished on concentrate and a 0.60-kg increase for lambs finished on forage (diet x slaughter weight, P < 0.03); however, the increased loin weight for lambs finished on concentrate was due largely to increased fat deposition. For lambs slaughtered at 77 kg, those finished on forage had more lean mass in the leg, loin, rack, and shoulder than those finished on concentrate, but lean mass in these cuts did not differ between diets for lambs slaughtered at 52 kg (diet x slaughter weight, P < 0.01). At the normal slaughter weight (52 kg), concentrate fed lambs had 50% more dissectible fat than forage-fed lambs, whereas at the heavy slaughter weight, a 79% greater amount of dissectible fat was observed for concentrate- vs. forage-fed lambs (diet x slaughter weight, P < 0.001). Lean and fat accretion rates were higher (P < 0.001) for concentrate-fed lambs than for forage-fed lambs. The lean-to-fat ratio of forage-fed lambs was higher (P < 0.001) than that of concentrate-fed lambs; however, forage finishing decreased accretion rates of all tissues compared with concentrate feeding, and these differences between forage and concentrate feeding were magnified at heavier slaughter weights. PMID- 15890812 TI - Prediction of retail product and trimmable fat yields from the four primal cuts in beef cattle using ultrasound or carcass data. AB - The most widely used system to predict percentage of retail product from the four primal cuts of beef is USDA yield grade. The purpose of this study was to determine whether routine ultrasound measurements and additional rump measurements could be used in place of the carcass measurements used in the USDA yield grade equation to more accurately predict the percentage of saleable product from the four primals. This study used market cattle (n = 466) consisting of Angus bulls, Angus steers, and crossbred steers. Live animal ultrasound measures collected within 7 d of slaughter were 1) scan weight (SCANWT); 2) 12th- to 13th-rib s.c. fat thickness (UFAT); 3) 12th- to 13th-rib LM area (ULMA); 4) s.c. fat thickness over the termination of the biceps femoris in the rump (URFAT; reference point); 5) depth of gluteus medius under the reference point (URDEPTH); and 6) area of gluteus medius anterior to the reference point (URAREA). Traditional carcass measures collected included 1) HCW; 2) 12th-to 13th-rib s.c. fat thickness (CFAT); 3) 12th- to 13th-rib LM area (CLMA); and 4) estimated percentage of kidney, pelvic, and heart fat (CKPH). Right sides of carcasses were fabricated into subprimal cuts, lean trim, fat, and bone. Weights of each component were recorded, and percentage of retail product from the four primals was expressed as a percentage of side weight. A stepwise regression was performed using data from cattle (n = 328) to develop models to predict percentage of retail product from the four primals based on carcass measures or ultrasound measures, and comparisons were made between the models. The most accurate carcass prediction equation included CFAT, CKPH, and CLMA (R2 = 0.308), whereas the most accurate live prediction equation included UFAT, ULMA, SCANWT, and URAREA (R2 = 0.454). When these equations were applied to a validation set of cattle (n = 138), the carcass equation showed R2 = 0.350, whereas the ultrasound data showed R2 = 0.460. Ultrasound measures in the live animal were potentially more accurate predictors of retail product than measures collected on the carcass. PMID- 15890813 TI - Effects of ractopamine on performance and composition of pigs phenotypically sorted into fat and lean groups. AB - Crossbred barrows (n = 144; 80 kg) from four farrowing groups were phenotypically selected into fat (FAT) and lean (LEAN) pens using ultrasound. The difference in 10th-rib fat depth between the LEAN and FAT groups was > or =0.5 cm. Within a farrowing group, pigs were assigned to pens (five pigs per pen and eight pens per phenotype) to equalize pen weight and fat depth. Pigs were fed a corn-soybean meal diet containing 19% CP, 1.0% added animal/vegetable fat, and 1.1% lysine (as fed basis). Half the pens received 10 ppm (as-fed basis) of ractopamine (RAC) during the 28-d finishing phase. At 7-d intervals, live weight and feed disappearance were recorded to calculate ADG, ADFI, and G:F, and 10th-rib fat depth and LM area were ultrasonically measured to calculate fat-free lean and fat and muscle accretion rates. During the first 7 d on feed, LEAN pigs fed RAC gained less (P < 0.05) than FAT pigs fed RAC or LEAN and FAT pigs fed the control diet (RAC x phenotype; P = 0.02); however, RAC did not (P > 0.25) affect ADG after the second, third, and fourth weeks, or over the entire 28-d feeding period. Although wk-2 and -3 ADG were higher (P < or = 0.03) in LEAN than in FAT pigs, phenotype did not (P = 0.08) affect overall ADG. Dietary RAC decreased (P < or = 0.05) ADFI over the 28-d feeding trial, as well as in wk 2, 3, and 4, but intake was not (P > 0.20) affected by phenotype. Neither RAC nor phenotype affected (P > 0.10) G:F after 7 d on trial; however, RAC improved (P < or = 0.04) wk-3, wk-4, and overall G:F. Lean pigs were more efficient (P < or = 0.05) in wk 2 and 3 and over the duration of the trial than FAT pigs. Ultrasound LM accretion (ULA) was not (P > or = 0.10) affected by RAC; however, LEAN pigs had greater (P < or = 0.02) ULA in wk 2 and 4 than FAT pigs. Although fat depth was lower (P < 0.01) in RAC-fed pigs than pigs fed the control diet, ultrasound fat accretion rate indicated that RAC-pigs deposited less (P = 0.04) fat only during wk 4. In addition, calculated fat-free lean (using ultrasound body fat, ULA, and BW) was increased (P < 0.05) in RAC pigs after 3 and 4 wk of supplementation. In conclusion, RAC enhanced the performance of finishing swine through decreased ADFI and increased G:F, whereas carcass lean was enhanced through decreases in carcass fat and increases in carcass muscling. PMID- 15890814 TI - Effect of pH and ionic strength on mu- and m-calpain inhibition by calpastatin. AB - The objectives of this study were to determine the extent to which pH and ionic strength influence mu- and m-calpain activity and the inhibition of calpains by calpastatin. Calpastatin, mu-calpain, and m-calpain were purified from at-death porcine semimembranosus. Mu-calpain or m-calpain (0.45 U) were incubated with the calpain substrate Suc-Leu-Leu-Val-Tyr-7-amino-4-methyl coumarin in the presence of calpastatin (0, 0.15, or 0.30 U of calpain inhibitory activity) under the following pH and ionic strength conditions: pH 7.5 and 165 mM NaCl or 295 mM NaCl; pH 6.5 and 165 mM NaCl or 295 mM NaCl; and pH 6.0 and 165 mM NaCl or 295 mM NaCl. The reactions were initiated with addition of 100 microM (mu-calpain) or 1 mM CaCl2 (m-calpain), and calpain activity was recorded at 30 and 60 min. Mu calpain had the greatest (P < 0.01) activity at pH 6.5 at each ionic strength. Higher ionic strength decreased mu-calpain activity (P < 0.01) at all pH conditions. Inhibition percent of mu-calpain by calpastatin was not affected by pH; however, it was influenced by ionic strength. Inhibition of mu-calpain by calpastatin was higher (P < 0.01) at 295 mM NaCl than at 165 mM NaCl when 0.3 units of calpastatin were included in the assay. Activity of m-calpain was greater (P < 0.01) at pH 7.5 than at pH 6.5. m-Calpain activity was not detected at pH 6.0. Inhibition of m-calpain was greater (P < 0.01) when 0.15 and 0.3 U calpastatin were added at pH 6.5 than 7.5 at 165 mM NaCl, whereas percentage inhibition of m-calpain was greater (P < 0.01) at 295 mM than 165 mM NaCl at pH 7.5 and 6.5. These observations provide new evidence that defines further the influence of pH decline and increased ionic strength on mu-calpain, m-calpain, and calpastatin activity, thereby helping to more accurately define a role for these enzymes in the process of postmortem tenderization. PMID- 15890815 TI - Estimates of heat stress relief needs for Holstein dairy cows. AB - Estimates of environmental heat stress are required for heat stress relief measures in cattle. Heat stress is commonly assessed by the temperature-humidity index (THI), the sum of dry and wet bulb temperatures. The THI does not include an interaction between temperature and humidity, although evaporative heat loss increases with rising air temperature. Coat, air velocity, and radiation effects also are not accounted for in the THI. The Holstein dairy cow is the primary target of heat stress relief, followed by feedlot cattle. Heat stress may be estimated for a variety of conditions by thermal balance models. The models consist of animal-specific data (BW, metabolic heat production, tissue and coat insulation, skin water loss, coat depth, and minimal and maximal tidal volumes) and of general heat exchange equations. A thermal balance simulation model was modified to adapt it for Holstein cows by using Holstein data for the animal characteristics in the model, and was validated by comparing its outputs to experimental data. Model outputs include radiant, convective, skin evaporative, respiratory heat loss and rate of change of body temperature. Effects of milk production (35 and 45 kg/d), hair coat depth (3 and 6 mm), air temperature (20 to 45 degrees C), air velocity (0.2 to 2.0 m/s), air humidity (0.8 to 3.9 kPa), and exposed body surface (100, 75, and 50%) on thermal balance outputs were examined. Environmental conditions at which respiratory heat loss attained approximately 50% of its maximal value were defined as thresholds for intermediate heat stress. Air velocity increased and humidity significantly decreased threshold temperatures, particularly at higher coat depth. The effect of air velocity was amplified at high humidity. Increasing milk production from 35 to 45 kg/d decreased threshold temperature by 5 degrees C. In the lying cow, the lower air velocity in the proximity of body surface and the smaller exposed surface markedly decrease threshold temperature. The large variation in thresholds due to environmental and animal factors justifies the use of thermal balance-based indices for estimating heat stress. Such an approach may make possible estimates of threshold temperatures at which heat stress relief is required for widely different cattle types and environmental situations. PMID- 15890816 TI - Effects of increasing temperatures on physiological changes in pigs at different relative humidities. AB - The effects of relative humidity (RH) and high ambient temperature (T) on physiological responses and animal performance were studied using 12 groups (10 gilts per group) in pens inside respiration chambers. The microclimate in the chamber was programmed so that T remained constant within a day. Each day, the T was increased by 2 degrees C from low (16 degrees C) to high (32 degrees C). Relative humidity was kept constant at 50, 65, or 80%. The pigs' average initial BW was 61.7 kg (58.0 to 65.5 kg), and their average ending BW was 70.2 kg (65.9 to 74.7 kg). Respiration rate (RR), evaporative water (EW), rectal temperature (RT), skin temperature (ST), voluntary feed intake (VFI), water-to-feed ratio (rW:F), heat production (HP), and ADG were analyzed. The animals had free access to feed and water. We determined the T above which certain animal variables started to change: the so-called inflection point temperature (IPt) or "upper critical temperature." The first indicator of reaction, RR, was in the range from 21.3 to 23.4 degrees C. Rectal temperature was a delayed indicator of heat stress tolerance, with IPt values ranging from 24.6 to 27.1 degrees C. For both these indicators the IPt was least at 80% RH (P < 0.05). Heat production and VFI were decreased above IPt of 22.9 and 25.5 degrees C, respectively (P < 0.001). For each degree Celsius above IPt, the VFI was decreased by 81, 99, and 106 g/(pig.d) in treatments 50, 65, and 80% RH, respectively. The ADG was greatest at 50% RH (P < 0.05). Ambient temperature strongly affects the pigs' physiological changes and performance, whereas RH has a relatively minor effect on heat stress in growing pigs; however, the combination of high T and high RH lowered the ADG in pigs. The upper critical temperature can be considered to be the IPt above which VFI decreased and RT then increased. Temperatures of the magnitude of both these IPt are regularly measured in commercial pig houses. We conclude that the upper critical temperatures for 60-kg, group-housed pigs fed ad libitum are between 21.3 and 22.4 degrees C for RR, between 22.9 and 25.5 degrees C for HP and VFI, and between 24.6 and 27.1 degrees C for RT. It is clear that different physiological and productive measurements of group-housed, growing-finishing pigs have different critical temperatures. PMID- 15890817 TI - Use of a crush-reducing device to decrease crushing of suckling piglets by sows. AB - Weanling pigs tend to avoid wind, and suckling piglets are thought to be more sensitive to wind than weanlings, owing to their thinner s.c. fat layer. We developed a crush-reducing device based on the anticipated behavior of suckling piglets toward wind and evaluated the performance of the device through field testing. The crush-reducing device consisted of six photo sensors, a controller, six solenoid valves, and an air compressor. In this study, 206 sows and their suckling piglets (Landrace xYorkshire) were investigated to ascertain the individual effects of several factors. Some of the newborn piglets were cross fostered before the crush-reducing device was implemented. Litter weights were measured on d 0 and 4 to determine the influence of the crush-reducing device on the weights of suckling piglets. The crushing of suckling piglets by sows was affected by season (P < 0.01) and litter size (P < 0.05), but not by the parity of the sows; however, the number of crushed piglets per litter was less (P < 0.01) in the crush-reducing device group (0.05 +/- 0.02 crushed piglets/litter) than in the control group (0.23 +/- 0.04 crushed piglets/litter), regardless of litter size or season. The BW gain of suckling piglets did not differ between the control and the crush-reducing device groups. Based on these results, the crush reducing device is expected to decrease the number of crushed piglets per litter without influencing the BW gain of suckling piglets, thereby greatly contributing to the productivity of pig breeders. PMID- 15890818 TI - Effect of administration of human chorionic gonadotropin after artificial insemination on concentrations of progesterone and conception rates in beef heifers. AB - The objective of this study was to determine whether administration of hCG approximately 5 d after AI would increase plasma progesterone concentrations and conception rates in beef heifers. Heifers from two locations (Location 1: n = 347, BW = 367 +/- 1.72 kg; Location 2: n = 246, BW = 408 +/- 2.35 kg) received melengestrol acetate (0.5 mg.heifer(-1).d(-1)) for 14 d and an injection of PGF2alpha (25 mg i.m.) 19 d later. Heifers were observed for estrus continuously during daylight from d 0 to 4.5 after PGF2alpha and artificially inseminated approximately 12 h after the onset of estrus. Half of the heifers inseminated at Location 1 were assigned randomly to receive an injection of hCG (3,333 IU i.m.) 8 d after PGF2alpha, and a blood sample was collected from all heifers 14 d after PGF2alpha for progesterone analysis. Half of the heifers inseminated at Location 2 were administered hCG on d 9 after PGF2alpha, and a blood sample was collected from all heifers 17 d after PGF2alpha. Heifers at Location 1 had a 94% synchronization rate, exhibited estrus 2.45 +/- 0.03 d after PGF2alpha, and received hCG 5.55 +/- 0.03 d after AI. Heifers at Location 2 had an 85% synchronization rate, exhibited estrus 2.69 +/- 0.03 d after PGF2alpha, and received hCG 6.31 +/- 0.03 d after AI. Progesterone concentrations were greater (P < 0.01) for hCG-treated heifers than for controls at both locations (8.6 vs. 4.6 ng/mL for treatment vs. control at Location 1, and 11.2 vs. 5.6 ng/mL for treatment vs. control at Location 2). Pregnancy status was determined by ultrasound approximately 50 d after AI. Conception rates (65 vs. 70% for treatment vs. control, respectively) did not differ at Location 1. Conception rates tended (P = 0.10) to be increased with hCG treatment at Location 2 (61 vs. 50% for treatment vs. control, respectively). A second experiment was conducted with 180 heifers at a third location to determine the effects of hCG administration 6 d after timed insemination at approximately 60 h after PGF2alpha in heifers synchronized as in Exp. 1. Pregnancy rate to timed AI did not differ between hCG-treated (62%) and control heifers (59%). Final pregnancy rate after timed AI and bull exposure (92%) was not affected by treatment. In summary, administration of hCG 5 to 6 d after AI did not improve conception or pregnancy rates at two out of three locations evaluated, suggesting insufficient progesterone is not a major factor contributing to early pregnancy failure in beef heifers. PMID- 15890819 TI - Six component intervals of nonproductive days by breeding-female pigs on commercial farms. AB - Of 105 swine herds using a production record system for breeding female pigs, 95 farms were used to analyze nonproductive female days (NPD), the six component intervals of NPD, and related measurements. The NPD was defined as the days when mated gilts and sows were neither gestating nor lactating, and it was calculated by summing the six component intervals in the average mated female inventory. The mean NPD was 57.9 d (SD = 20.5), and the proportions of six component intervals of gilt first-mating-to-pregnancy interval, gilt first-mating-to-culling interval, unmated weaning-to-culling interval, weaning-to-first-mating interval, sow first-mating-to-pregnancy interval, and sow first-mating-to-culling interval were 9.24, 7.82, 6.85, 27.9, 18.9, and 29.3%, respectively. Farms in the upper 25th percentile of the ranking for number of pigs weaned.mated female(-1).yr(-1) were designated as 25 high-performing farms. The remaining farms were designated as an ordinary farm group for comparisons. High-performing farms had 21.1 d fewer NPD, and five of the six component intervals were lower compared with the ordinary farms (P < 0.05). Regression analyses indicated that the number of litters.mated female(-1).yr(-1) increased by 0.07 in both farm groups as NPD decreased every 10 d. Fewer NPD were correlated with a higher percentage of multiple matings during estrus (P < 0.05) but were not correlated with removal risk and replacement risk in both farm groups. The average parity of culled females was negatively correlated with NPD in the ordinary farm group, and the average farrowed parity was positively correlated with NPD in the high-performing farm group (P < 0.01). Decreasing each component interval of the NPD six components is critical to increasing herd productivity. A high percentage of multiple matings during estrus and appropriate culling management may be key factors to decrease NPD. PMID- 15890820 TI - Water intake and wastage at nipple drinkers by growing-finishing pigs. AB - Three experiments were conducted to assess water intake, water wastage, and a means to decrease water wastage by growing-finishing pigs from nipple drinkers. In Exp. 1, 48 pigs were studied during two periods (average BW = 53 and 72 kg for Period 1 and 2, respectively). Water disappearance and wastage were determined for 4 d, while nipple drinkers were set at 50 mm above the shoulder height of the smallest pig in the pen (recommended heights), with flow rates at 700 mL/min during Period 1, and 1,000 mL/min during Period 2. Water intake rate was assessed at two nipple flow rates, approximately 650 and 1,300 mL/min during the Period 1, and 1,000 and 2,000 mL/min during the Period 2. The average water intake was 4.01 and 5.38 +/- 0.19 L.pig(-1).d(-1) during Periods 1 and 2, respectively (P < 0.01). Water wastage as a percentage of water disappearance was similar between the two periods (25.8 and 27.0 +/- 1.9% for Periods 1 and 2, respectively). Water intake rate was 467 and 795 mL/min (+/-34.2; P < 0.01) during Period 1, and 722 and 1,422 mL/min (+/-80.0; P < 0.01) during Period 2, at the lower and higher flow rates, respectively. In Exp. 2, 32 pigs were used in a 2 x 2 factorial design to determine effects of nipple heights (recommended vs. unadjusted = 330 mm) and flow rates (500 vs. 1,000 mL/min) on water intake and wastage. Water wastage was increased (P < 0.01) on the unadjusted vs. recommended nipple height, and the higher flow rate also resulted in greater wastage (P = 0.03) compared with the lower rate. In Exp. 3, water disappearance and manure output in 16 pens of 18 pigs per pen were monitored for 12 wk (average initial BW = 32 kg) using four drinker treatments (bowl drinker, nipple drinker at recommended heights, an unadjusted nipple set at 480 mm, and high nipple drinker height of 730 mm with a step underneath). For pigs on the high nipple drinker, the average water disappearance and manure output did not differ from those of the pigs on the recommended nipple heights and bowl drinker, but these measurements were 15 and 12% lower, respectively, than for the pigs on the low nipple drinker. The results indicate that growing-finishing pigs can maintain adequate water intake from a variety of drinker types and management. Water wastage can be controlled through drinker management. PMID- 15890821 TI - A model of fescue toxicosis: responses of rats to intake of endophyte-infected tall fescue. AB - A study was conducted to develop a model for fescue toxicosis using rats fed a diet containing endophyte-infected tall fescue seed (E+). Rats implanted with telemetric transmitters to continuously monitor core body temperature (Tc) and activity were housed at thermoneutrality (21 degrees C) and were fed a diet containing endophyte-free fescue seed (E-). After 2 wk, they were assigned to either E+ or E- diets and initially maintained at thermoneutrality (preheat) for 8 d. They were then exposed to heat stress (31 degrees C) for 22 d, followed by 1 wk of recovery at thermoneutrality (post-heat). Body weight and feed intake were measured daily. Rats receiving the E+ diet showed decreased feed intake (P = 0.001) and weight gains (P = 0.003) during the preheat period. The decrease in Tc from the pre-treatment level was greater in E+ than in E- rats during the preheat (P = 0.001) and postheat (P = 0.001) periods. With heat stress, both groups showed parallel decreases in feed intake. The increase in Tc from pre-heat to heat conditions was greater in E+ vs. E- rats (P = 0.001). Activity level was lower in E+ than in E-rats during heat stress (P = 0.009) and postheat (P = 0.037) periods. These results show that the rat model for fescue toxicosis is extremely useful because many of the observed responses to E+ diet are similar to those noted for cattle, and additional variables that are difficult to measure in cattle, such as activity, can be easily evaluated. PMID- 15890822 TI - Influence of sward height, daily timing of concentrate supplementation, and restricted time for grazing on forage utilization by lactating beef cows. AB - To establish the effect of sward height, concentrate feeding time, and restricted time for grazing on forage utilization by grazing cattle, 32 crossbred beef (24 Angus and eight Hereford) cows (632 kg BW) and calves (104 kg BW) were grouped by weight and calving date. They were assigned randomly to two sward height treatments (4 to 8 or 8 to 12 cm), replicated four times. The herbage comprised mainly Kentucky bluegrass, orchardgrass, some forbs, and white clover. The cows were restricted to 12 h/d grazing (0700 to 1900) or unrestricted to 24 h/d grazing and fed a concentrate supplement (4.1 kg DM.cow(-1).d(-1), approximately 0.65% of BW or 33% of total DMI) either at 0700 or 1800. The experiment was repeated over three 15-d periods in May, June/July, and August 2000. The herbage on high sward height pasture was higher (P = 0.06) in NDF and ADF and lower (P < 0.01) in CP than low sward height herbage. For cows restricted to 12 h/d grazing, supplementing at 0700 as opposed 1800 resulted in greater (P = 0.04) forage DMI (8.6 vs. 8.1 kg/d), whereas cows that were unrestricted showed little change (8.2 kg/d at 0700 vs. 8.4 kg/d at 1800). Supplementing at 1800 as opposed to 0700 resulted in greater (P = 0.03) herbage DM digestibility (67.7 vs. 64.5%) for cows on high sward height, whereas cows on low sward height exhibited minimal differences (65.4% at 1800 vs. 66.3% at 0700). Cows restricted to 12 h/d grazing and supplemented at 0700 as opposed to 1800 resulted in greater (P = 0.06) digestible DMI (5.0 vs. 4.7 kg/d), whereas unrestricted cows exhibited the opposite response (4.6 kg/d digestible DMI at 0700 vs. 4.9 kg/d at 1800). Supplementing at 1800 as opposed to 0700 increased the time spent grazing to a greater (P = 0.09) extent for restricted than for unrestricted cows. When forage availability or grazing time was limiting (due to a low forage allowance and restricted access to forage, respectively) supplementing concentrates at 0700 resulted in greater forage utilization and intake rate because of increased forage DMI, DM digestibility, and digestible DMI. However, when forage or grazing time was not limiting, supplementing concentrates at 1800 resulted in greater forage utilization because of increased forage DM digestibility. PMID- 15890823 TI - Effect of cutting height and genetics on composition, intake, and digestibility of corn silage by beef heifers. AB - This research was conducted to determine the effect of corn genetics and cutting height on the composition and nutritive characteristics of corn silage. An in situ study involving eight commercially available corn hybrids indicated main effects and interactions (P < 0.01) of hybrid and cutting height on NDF, ADF, and starch content and on in situ DM and NDF degradablility. Four ruminally cannulated Angus heifers (initial BW = 378 +/- 3 kg) were used in a 4 x 4 Latin square digestion experiment with a 2 x 2 factorial treatment arrangement. Main effects and interactions of hybrid (Pioneer Hi-Bred Int., Inc., hybrids 3335 and 3223) and cutting height (LO = 20.3 cm, and HI = 61 cm) were evaluated. Dietary treatment consisted of 40% chopped alfalfa hay and 60% corn silage. Although corn silage hybrids used were of equivalent maturity at harvest (60% milkline), 3335 treatments had 37.8% starch and 34.8% NDF, whereas 3223 treatments had 33.7% starch and 38.6% NDF. The LO treatments averaged 3.1 percentage units greater in NDF and 3.45 percentage units less in starch content than the HI treatments. Intake of DM was greater for heifers fed 3335-HI than 3335-LO; however, DMI was greater by heifers fed 3223-LO than 3223-HI (hybrid x cutting height interaction, P < 0.05). Starch intake was greater (P < 0.05) and NDF intake was less (P < 0.05) by heifers fed HI vs. LO and fed 3335 vs. 3223 dietary treatments. Digestibility of DM, starch, and NDF was greater (P < 0.05) by heifers fed 3223 than 3335 dietary treatments, but digestibility differences were not observed (P > 0.10) between cutting heights. Rate of in situ DM and starch degradability was not affected (P > 0.10) by hybrid or cutting height; however DM degradability was greater (P < 0.05) for HI than LO corn silage substrates at 8, 16, and 24 h of incubation. Rate of NDF degradability tended (P = 0.08) to be greater for 3223 than for 3335, and for LO compared with HI corn silage. Degradability of NDF was greater (P < 0.05) for 3223 compared with 3335 substrates at 24, 36, and 48 h of incubation. These data suggest fiber may not be an accurate measure of corn silage quality. Whereas cutting height affected chemical composition, we observed genetics to have a greater effect on corn silage quality. PMID- 15890824 TI - Prion biology relevant to bovine spongiform encephalopathy. AB - Bovine spongiform encephalopathy (BSE) and chronic wasting disease (CWD) of deer and elk are a threat to agriculture and natural resources, as well as a human health concern. Both diseases are transmissible spongiform encephalopathies (TSE), or prion diseases, caused by autocatalytic conversion of endogenously encoded prion protein (PrP) to an abnormal, neurotoxic conformation designated PrPsc. Most mammalian species are susceptible to TSE, which, despite a range of species-linked names, is caused by a single highly conserved protein, with no apparent normal function. In the simplest sense, TSE transmission can occur because PrPsc is resistant to both endogenous and environmental proteinases, although many details remain unclear. Questions about the transmission of TSE are central to practical issues such as livestock testing, access to international livestock markets, and wildlife management strategies, as well as intangible issues such as consumer confidence in the safety of the meat supply. The majority of BSE cases seem to have been transmitted by feed containing meat and bone meal from infected animals. In the United Kingdom, there was a dramatic decrease in BSE cases after neural tissue and, later, all ruminant tissues were banned from ruminant feed. However, probably because of heightened awareness and widespread testing, there is growing evidence that new variants of BSE are arising "spontaneously," suggesting ongoing surveillance will continue to find infected animals. Interspecies transmission is inefficient and depends on exposure, sequence homology, TSE donor strain, genetic polymorphism of the host, and architecture of the visceral nerves if exposure is by an oral route. Considering the low probability of interspecies transmission, the low efficiency of oral transmission, and the low prion levels in nonnervous tissues, consumption of conventional animal products represents minimal risk. However, detection of rare events is challenging, and TSE literature is characterized by subsequently unsupported claims of species barriers or absolute tissue safety. This review presents an overview of TSE and summarizes recent research on pathogenesis and transmission. PMID- 15890825 TI - Structure and function of the notochord: an essential organ for chordate development. AB - The notochord is the defining structure of the chordates, and has essential roles in vertebrate development. It serves as a source of midline signals that pattern surrounding tissues and as a major skeletal element of the developing embryo. Genetic and embryological studies over the past decade have informed us about the development and function of the notochord. In this review, I discuss the embryonic origin, signalling roles and ultimate fate of the notochord, with an emphasis on structural aspects of notochord biology. PMID- 15890826 TI - BOP, a regulator of right ventricular heart development, is a direct transcriptional target of MEF2C in the developing heart. AB - The vertebrate heart is assembled during embryogenesis in a modular manner from different populations of precursor cells. The right ventricular chamber and outflow tract are derived primarily from a population of progenitors known as the anterior heart field. These regions of the heart are severely hypoplastic in mutant mice lacking the myocyte enhancer factor 2C (MEF2C) and BOP transcription factors, suggesting that these cardiogenic regulatory factors may act in a common pathway for development of the anterior heart field and its derivatives. We show that Bop expression in the developing heart depends on the direct binding of MEF2C to a MEF2-response element in the Bop promoter that is necessary and sufficient to recapitulate endogenous Bop expression in the anterior heart field and its cardiac derivatives during mouse development. The Bop promoter also directs transcription in the skeletal muscle lineage, but only cardiac expression is dependent on MEF2. These findings identify Bop as an essential downstream effector gene of MEF2C in the developing heart, and reveal a transcriptional cascade involved in development of the anterior heart field and its derivatives. PMID- 15890827 TI - "In the nation's compelling interest". PMID- 15890828 TI - Can women at term self-diagnose premature spontaneous rupture of membranes? PMID- 15890829 TI - The travesty of choosing after positive prenatal diagnosis. AB - OBJECTIVE: To integrate the findings of qualitative studies of expectant parents receiving positive prenatal diagnosis. DATA SOURCES: Seventeen published and unpublished reports appearing between 1984 and 2001 and retrieved between December of 2002 and March of 2003. The electronic databases searched include Academic Search Elite, AIDS Information Online (AIDSLINE), Anthropological Index Online, Anthropological Literature, Black Studies, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Digital Dissertations, Dissertation Abstracts Index (DAI), Educational Resource Information Center (ERIC), MEDLINE, PsycInfo, Public Affairs Information Service (PAIS), PubMed, Social Science Abstracts (SocSci Abstracts), Social Science Citation Index, Social Work Abstracts, Sociological Abstracts (Sociofile), Women's Resources International, and Women's Studies. STUDY SELECTION: Qualitative studies involving expectant parents living in the United States of any race, ethnicity, nationality, or class who learned during any time in pregnancy of any fetal impairment by any means of diagnosis were eligible for inclusion. DATA EXTRACTION: Metasummary techniques, including the calculation of frequency effect sizes, were used to aggregate the findings. Metasynthesis techniques, including constant comparison analysis and the reciprocal translation of concepts, were used to interpret the findings. DATA SYNTHESIS: The topical emphasis in the findings is on the termination of pregnancy following positive diagnosis. The thematic emphasis is on the dilemmas of choice and decision making. Positive prenatal diagnosis was for couples an experience of chosen losses and lost choices. Couples managed information to minimize stigmatization and cognitive dissonance. CONCLUSIONS: Existing guidelines for caring for couples after perinatal losses must accommodate the chosen loss experientially defining positive prenatal diagnosis. PMID- 15890830 TI - The effect of unrestricted oral carbohydrate intake on labor progress. AB - OBJECTIVE: To determine if unrestricted oral carbohydrate intake during labor reduced the incidence of dystocia in low-risk nulliparous women. DESIGN AND SETTING: A randomized clinical trial at a university-affiliated hospital in southeastern Ontario. Low-risk nulliparous women were randomized between 30 and 40 weeks gestation to either an intervention or usual care group. INTERVENTION: Women in the intervention group received, prenatally, guidelines about food and fluid intake during labor and were encouraged to eat and drink as they pleased during labor. Women in the usual care group received no prelabor information and were restricted to ice chips and water during labor in the hospital. MAIN OUTCOME MEASURE: The incidence of dystocia, defined as a cervical dilatation rate of less than 0.5 cm/hr for a period of 4 hrs after a cervical dilatation of 3 cm. RESULTS: Three hundred twenty-eight women were randomized to the intervention (n = 163) or usual care (n = 165) groups. Women in the intervention group reported a significantly different pattern of oral intake during early labor in the hospital (chi(2) = 40.7, p < .001). The incidence of dystocia was 36% (n = 58) in the intervention group and 44% (n = 72) in the usual care group and was not significantly different (OR = 0.71, 95% CI = 0.46, 1.11). There were no significant differences in the other secondary outcomes or in the incidence of adverse maternal or neonatal complications. CONCLUSION: Eating and drinking early in labor had no significant impact on the incidence of dystocia and/or adverse maternal or neonatal outcomes. PMID- 15890831 TI - Reliability of reporting of self-monitoring of blood glucose in pregnant women. AB - OBJECTIVE: To determine the accuracy of reporting of self-monitored blood glucose in pregnant women with diabetes. DESIGN: A descriptive study. Patient-recorded logs of self-monitored blood glucose values were compared to meter memory values. SETTING AND PARTICIPANTS: A convenience sample of 85 pregnant women with pregestational and gestational diabetes enrolled in a perinatal diabetes program in an urban teaching hospital. RESULTS: Accuracy significantly differed by diabetes type (p = .015). Women with type 1 diabetes did not accurately record on average 36.7% of blood glucose values as compared to 8.5% of type 2, 21.2% of GDMA(1) (gestational diabetes mellitus, diet controlled), and 23.4% of GDMA(2) (gestational diabetes mellitus, insulin controlled). Age positively affected accuracy, but accuracy was not affected by marital status, educational background, or duration of diabetes. Statistical significance was shown between values from women with private health insurance and women with Medicaid. Eighty percent of the sample overreported by adding phantom values in the logbook, which did not differ by diabetes type. Approximately 70% underreported by not logging values in the meter memory that was statistically significant by diabetes type. CONCLUSION: Assessment of the reliability of self-monitored blood glucose results from pregnant women with diabetes is recommended owing to a significant degree of falsification. PMID- 15890832 TI - Neurobehavioral functioning and breastfeeding behavior in the newborn. AB - OBJECTIVE: To determine whether central nervous system functioning has an effect on the normal, term infant's ability to breastfeed in the first day following birth. DESIGN: Breastfeeding behaviors and neurobehaviors were evaluated at birth and at 24 hrs of age in two groups of neonates. One group of neonates was born to mothers who received epidural analgesia during labor, and one group was born to mothers who received no pain medication. Breastfeeding behavior was evaluated using the Preterm Infant Breastfeeding Behavior Scale, and the infant's neurobehaviors were evaluated using the Neurologic and Adaptive Capacity Score. PARTICIPANTS: Fifty-six breastfeeding mother-newborn dyads. All mothers were healthy multiparae who gave birth vaginally to normal, full-term, healthy newborns. MAIN OUTCOME MEASURES: Newborns were observed for rooting, latch-on, sucking, swallowing, activity state, and neurobehavior. RESULT: Analysis of the data indicated that the higher the infant scored in relation to neurobehavioral functioning, the higher the infant scored on breast-feeding behaviors. CONCLUSION: When determining physiologically what is important for the infant to successfully latch on and feed, it appears that an intact and functioning central nervous system may be one of the crucial elements. PMID- 15890833 TI - Sleep-wake cycles, social rhythms, and sleeping arrangement during Japanese childbearing family transition. AB - OBJECTIVE: To examine how sleep-wake cycles and social rhythms of Japanese parents are related to their sleeping arrangements before and after the birth of their first child. DESIGN: Prospective longitudinal, time-series methods. SETTING: Participants' homes in four geographical areas of Japan. PARTICIPANTS: Convenience sample of 101 Japanese first-time parent couples who completed antepartum and postpartum questionnaires. MAIN OUTCOME MEASURES: 24-hr sleep-wake rhythm from 7-day logs at 32 to 36 weeks gestation and 4 to 5 weeks postpartum and daily social rhythm from Monk's Social Rhythm Metric. RESULTS: Mothers' total sleep time and activity level decreased after birth. Social rhythms became less regular for mothers and more consistent and regular for fathers. Although sleeping arrangement had no effect on parents' daily social rhythms, it had a significant effect on the 24-hr sleep-wake rhythm. Regularity of sleep-wake rhythms was more prominent for fathers sleeping with mother and baby than for fathers sleeping alone. CONCLUSIONS: Couples having their first child face great changes in their sleep-wake cycles and social rhythms. Nurses can counsel expectant parents to think about their sleeping habits and social activities and assist them in making informed decisions about the best sleeping arrangement during transition to parenthood. PMID- 15890834 TI - The grounded theory method and maternal-infant research and practice. AB - Perinatal nurses have found qualitative methods helpful in capturing the social, cultural, and relational aspects of their work with women and their families. This article describes the components of the grounded theory method and illustrates each with examples from recent maternal-infant studies. Grounded theory is an inductive method of qualitative analysis that generates theory from data. The grounded theory method includes several key strategies such as theoretical sensitivity, theoretical sampling, constant comparison, increasingly abstract consideration of the data, and discovery of a core variable or basic social process that describes the pattern of the phenomenon under study. This method may strengthen qualitative nursing studies because of its generalizability and applicability across settings. By remaining grounded in and connected to the data, grounded theory can be adapted to many contexts, making it useful to the practicing nurse. Knowledge generated from grounded theory has great potential to assist us in our inquiry into the needs of mothers, fathers, infants, and their families. PMID- 15890835 TI - Antenatal events causing neonatal brain injury in premature infants. AB - Understanding possible causes of neonatal brain injury is important for perinatal nurses because neonatal brain injury predicts subsequent infant mortality and morbidity in the premature infant. The pathogenesis of the brain injury, germinal matrix/intraventricular hemorrhage and white matter damage, is usually related to a hypoxic event. The hypoxic event may occur in utero, resulting from various conditions, such as maternal infection, maternal alcohol consumption, maternal smoking, placental bleeding disorders, maternal hypercoagulability conditions, metabolic disorders (diabetes and hyperthyroidism), and oligohydramnios. Strategies for prevention beginning before and in pregnancy are needed. PMID- 15890836 TI - Vitamin D and the breastfed infant. AB - For infants and children, vitamin D deficiency causes a bone-deforming disease known as rickets. As breastfeeding rates have increased, so have the incidences of rickets. The current recommendation from the American Academy of Pediatrics, to supplement all breastfed infants with vitamin D, is controversial. The role of the nurse is to understand the vitamin D dilemma, promote breastfeeding, and prevent vitamin D deficiency rickets. PMID- 15890837 TI - Providing options counseling for women with unintended pregnancies. AB - Nearly half of the pregnancies in the United States are unintended, and nurses frequently encounter women with unintended pregnancies in the clinical setting. Nurses may be responsible for counseling these women on how to explore and decide between the options of parenting, adoption, and abortion. To date, little has been written about options counseling, despite the fact that unintended pregnancy is one of the most common health conditions that women face. This article presents epidemiological data on prevalence, outcomes, consequences, causes, and prevention of unintended pregnancy. It examines the conflicts that nurses may experience when providing care to women with unintended pregnancies and discusses the intersection of professional responsibilities and personal values. Finally, the article presents general strategies and specific suggested steps for providing options counseling. PMID- 15890838 TI - Bridging health care disparities: addressing unmet needs of women of color. PMID- 15890839 TI - Factors influencing mammography screening of Chinese American women. AB - OBJECTIVE: To explore factors influencing breast cancer screening behavior among Chinese women residing in the United States. DESIGN: A descriptive study guided by the health belief model adapted for Chinese American women. An English language questionnaire was modified, translated, and pretested before use in the study sample. Data were analyzed using descriptive and multivariate analysis techniques. SETTING: An urban area in Michigan in 2001. PARTICIPANTS: A consecutive nonprobability sample of 206 Chinese American women age 40 and older. MAIN OUTCOME MEASURE: The percentage of women age 40 and older who received a mammogram in the past year. RESULTS: Access to health care, perceived barriers to mammography screening, need for breast health care, and information-seeking behavior had direct effects on Chinese American women's mammography screening utilization. Cultural affiliation had an indirect effect on breast cancer screening behavior, moderated through access to health care. The variance in mammography screening explained by these factors was 51%. CONCLUSION: Effective strategies for promoting breast cancer screening among Chinese American women should address ways to improve information-seeking behaviors and access to health care. Cultural affiliation and beliefs should be considered when counseling Chinese American women regarding breast cancer screening. PMID- 15890840 TI - A multicultural and multilingual outreach program for cervical and breast cancer screening. AB - Partners in Health: The Breast and Cervical Health Cooperative is a New Orleans based community health intervention program designed to increase the access of underserved multicultural, multiethnic, and multilingual women to cervical and breast cancer health education and screening. This article describes innovative approaches used over the past 4 years to successfully link the services of a university cancer center to women in the community through a culturally sensitive outreach program. In addition to cancer screening, health referrals were made for many other diverse medical/social complications such as hypertension, obesity, and violence. The program initially targeted African American women but during the past year has successfully integrated Latinas. Key to the program's success was the collaboration of community lay health educators, community churches, advanced practice nurses, and public health programs. This article provides an overview of program strategies and outcomes and discusses multicultural and multilingual issues. PMID- 15890841 TI - cAMP and extracellular signal-regulated kinase signaling in response to d amphetamine and methylphenidate in the prefrontal cortex in vivo: role of beta 1 adrenoceptors. AB - d-Amphetamine and methylphenidate are widely used in the treatment of attention deficit/hyperactivity disorder. Both drugs increase extracellular norepinephrine and dopamine in the prefrontal cortex, where they are believed to exert their therapeutic effects. However, the molecular mechanisms underlying their action are poorly understood. To investigate the intracellular signaling pathways activated by d-amphetamine and methylphenidate in the prefrontal cortex in vivo in mice, we measured the cAMP-dependent Ser845 phosphorylation of alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid receptor GluR1 subunit and the active form of extracellular signal-regulated kinase (ERK). Administration of d amphetamine (5-10 mg/kg) or methylphenidate (10-20 mg/kg) increased phosphorylation of GluR1. Basal and d-amphetamine-induced GluR1 phosphorylation was reduced by propranolol, a general beta-adrenoceptor antagonist, and betaxolol, a beta1-antagonist, but not by (+/-)-1-[2,3-(dihydro-7-methyl-1H-inden 4-yl)oxy]-3-[(1-methylethyl)amino]-2-butanol (ICI-118,515), a beta2-antagonist. The effect of methylphenidate was also blocked by propranolol and betaxolol. The d-amphetamine effect was slightly potentiated by prazosin, an alpha1-adrenoceptor antagonist, and mimicked by yohimbine, an alpha2 antagonist. Blockade of dopamine or N-methyl-d-aspartate (NMDA) receptors or serotonin depletion had no effect on d-amphetamine-induced GluR1 phosphorylation. d-amphetamine but not methylphenidate increased ERK phosphorylation. This effect required multiple signaling pathways because it was blocked by beta1- and alpha1-adrenoceptor antagonists, by dizocilpine maleate (MK801), an NMDA antagonist, and by serotonin depletion. In contrast, blockade of dopamine receptors had no effect on d amphetamine-induced ERK phosphorylation. Propranolol and betaxolol increased the hyperlocomotion produced by d-amphetamine and methylphenidate. Thus, both d amphetamine and methylphenidate potently activate the cAMP pathway in the prefrontal cortex through beta1-adrenergic receptors. This activation could have behavioral consequences and contribute to the treatment of attention deficit/hyperactivity disorder. PMID- 15890842 TI - Orphanin FQ/nociceptin potentiates [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin-Induced mu-opioid receptor phosphorylation. AB - In this study, we investigate the molecular mechanisms by which acute orphanin FQ/nociceptin (OFQ/N), acting through the nociceptin opioid peptide (NOP) receptor, desensitizes the mu-opioid receptor. We described previously the involvement of protein kinase C and G-protein-coupled receptor kinases (GRK) 2 and 3 in OFQ/N-induced mu receptor desensitization. Because phosphorylation of the mu receptor triggers the successive regulatory mechanisms responsible for desensitization, such as receptor uncoupling, internalization, and down regulation, we investigated the ability of OFQ/N to modulate [d-Ala(2),N-Me Phe(4),Gly(5)-ol]-enkephalin (DAMGO)-induced mu receptor phosphorylation in BE(2) C human neuroblastoma cells transfected with epitope-tagged mu receptors. OFQ/N treatment (100 nM, 60 min) potentiated DAMGO-induced mu receptor phosphorylation; inhibition of GRK2 or protein kinase C concomitant with OFQ/N treatment blocked the OFQ/N-mediated increase in DAMGO-induced phosphorylation. Inclusion of the NOP antagonist peptide III-BTD during OFQ/N pretreatment blocked the potentiation of DAMGO-induced phosphorylation by OFQ/N, which is consistent with the potentiation being mediated via actions of the NOP receptor. In addition, in cells expressing mu receptors in which the GRK-mediated phosphorylation site Ser(375) was mutated to alanine, OFQ/N treatment failed to potentiate DAMGO induced mu receptor phosphorylation and failed to desensitize the mu receptor. However, DAMGO-induced mu receptor phosphorylation and OFQ/N-induced mu receptor desensitization occurred in cells expressing mu receptors lacking non-GRK phosphorylation sites. These data suggest that OFQ/N binds to NOP receptors and activates protein kinase C, which then increases the ability of GRK2 to phosphorylate the agonist-occupied mu receptor, heterologously regulating homologous mu receptor desensitization. PMID- 15890843 TI - Tubulin polyglutamylase enzymes are members of the TTL domain protein family. AB - Polyglutamylation of tubulin has been implicated in several functions of microtubules, but the identification of the responsible enzyme(s) has been challenging. We found that the neuronal tubulin polyglutamylase is a protein complex containing a tubulin tyrosine ligase-like (TTLL) protein, TTLL1. TTLL1 is a member of a large family of proteins with a TTL homology domain, whose members could catalyze ligations of diverse amino acids to tubulins or other substrates. In the model protist Tetrahymena thermophila, two conserved types of polyglutamylases were characterized that differ in substrate preference and subcellular localization. PMID- 15890844 TI - Enols are common intermediates in hydrocarbon oxidation. AB - Models for chemical mechanisms of hydrocarbon oxidation rely on spectrometric identification of molecular structures in flames. Carbonyl (keto) compounds are well-established combustion intermediates. However, their less-stable enol tautomers, bearing OH groups adjacent to carbon-carbon double bonds, are not included in standard models. We observed substantial quantities of two-, three-, and four-carbon enols by photoionization mass spectrometry of flames burning representative compounds from modern fuel blends. Concentration profiles demonstrate that enol flame chemistry cannot be accounted for purely by keto-enol tautomerization. Currently accepted hydrocarbon oxidation mechanisms will likely require revision to explain the formation and reactivity of these unexpected compounds. PMID- 15890845 TI - Climate change and distribution shifts in marine fishes. AB - We show that the distributions of both exploited and nonexploited North Sea fishes have responded markedly to recent increases in sea temperature, with nearly two-thirds of species shifting in mean latitude or depth or both over 25 years. For species with northerly or southerly range margins in the North Sea, half have shown boundary shifts with warming, and all but one shifted northward. Species with shifting distributions have faster life cycles and smaller body sizes than nonshifting species. Further temperature rises are likely to have profound impacts on commercial fisheries through continued shifts in distribution and alterations in community interactions. PMID- 15890846 TI - Living laboratory in peril. PMID- 15890847 TI - Fusion reactor. ITER rivals agree to terms; site said to be Cadarache. PMID- 15890848 TI - Stem cell research. California institute picks city by the bay. PMID- 15890849 TI - NASA astronomy. New space telescope may be scaled back. PMID- 15890850 TI - Microbiology. Global spread of leprosy tied to human migration. PMID- 15890851 TI - DOE weapons lab. Los Alamos appoints interim director. PMID- 15890853 TI - Sensitive technology. Schools fear impact of proposed license changes. PMID- 15890852 TI - Ecology. Fish moved by warming waters. PMID- 15890855 TI - Gamma ray astronomy. Signs point to neutron-star crash. PMID- 15890854 TI - Microbiology. Detente declared on NIH biodefense funding. PMID- 15890857 TI - Research funding. Centers of attention: NSF takes fresh look at their proliferation. PMID- 15890856 TI - Clinical research. Advocating, the clinical way. PMID- 15890858 TI - Neuroscience. Reflecting on another's mind. PMID- 15890859 TI - An open mind is a Trojan horse? PMID- 15890860 TI - Terrestriality and tool use. PMID- 15890861 TI - Consensus about climate change? PMID- 15890862 TI - Interpreting correlation as causation? PMID- 15890863 TI - Public health. Increasing international gaps in health-related publications. PMID- 15890864 TI - Ecology. 25 years of ecological change at Mount St. Helens. PMID- 15890865 TI - Atmospheric science. Rethinking Earth's early atmosphere. PMID- 15890866 TI - Structural biology. Flipping lipids: is the third time the charm? PMID- 15890868 TI - Geophysics. Past and future earthquakes on the San Andreas Fault. PMID- 15890867 TI - Evolution. Did early humans go north or south? PMID- 15890870 TI - Intensive Titan exploration begins. AB - The Cassini Orbiter spacecraft first skimmed through the tenuous upper atmosphere of Titan on 26 October 2004. This moon of Saturn is unique in our solar system, with a dense nitrogen atmosphere that is cold enough in places to rain methane, the feedstock for the atmospheric chemistry that produces hydrocarbons, nitrile compounds, and Titan's orange haze. The data returned from this flyby supply new information on the magnetic field and plasma environment around Titan, expose new facets of the dynamics and chemistry of Titan's atmosphere, and provide the first glimpses of what appears to be a complex, fluid-processed, geologically young Titan surface. PMID- 15890871 TI - Cassini radar views the surface of Titan. AB - The Cassini Titan Radar Mapper imaged about 1% of Titan's surface at a resolution of approximately 0.5 kilometer, and larger areas of the globe in lower resolution modes. The images reveal a complex surface, with areas of low relief and a variety of geologic features suggestive of dome-like volcanic constructs, flows, and sinuous channels. The surface appears to be young, with few impact craters. Scattering and dielectric properties are consistent with porous ice or organics. Dark patches in the radar images show high brightness temperatures and high emissivity and are consistent with frozen hydrocarbons. PMID- 15890872 TI - The Cassini UVIS stellar probe of the Titan atmosphere. AB - The Cassini Ultraviolet Imaging Spectrometer (UVIS) observed the extinction of photons from two stars by the atmosphere of Titan during the Titan flyby. Six species were identified and measured: methane, acetylene, ethylene, ethane, diacetylene, and hydrogen cyanide. The observations cover altitudes from 450 to 1600 kilometers above the surface. A mesopause is inferred from extraction of the temperature structure of methane, located at 615 km with a temperature minimum of 114 kelvin. The asymptotic kinetic temperature at the top of the atmosphere determined from this experiment is 151 kelvin. The higher order hydrocarbons and hydrogen cyanide peak sharply in abundance and are undetectable below altitudes ranging from 750 to 600 km, leaving methane as the only identifiable carbonaceous molecule in this experiment below 600 km. PMID- 15890873 TI - Ion neutral mass spectrometer results from the first flyby of Titan. AB - The Cassini Ion Neutral Mass Spectrometer (INMS) has obtained the first in situ composition measurements of the neutral densities of molecular nitrogen, methane, molecular hydrogen, argon, and a host of stable carbon-nitrile compounds in Titan's upper atmosphere. INMS in situ mass spectrometry has also provided evidence for atmospheric waves in the upper atmosphere and the first direct measurements of isotopes of nitrogen, carbon, and argon, which reveal interesting clues about the evolution of the atmosphere. The bulk composition and thermal structure of the moon's upper atmosphere do not appear to have changed considerably since the Voyager 1 flyby. PMID- 15890874 TI - Energetic neutral atom emissions from Titan interaction with Saturn's magnetosphere. AB - The Cassini Magnetospheric Imaging Instrument (MIMI) observed the interaction of Saturn's largest moon, Titan, with Saturn's magnetosphere during two close flybys of Titan on 26 October and 13 December 2004. The MIMI Ion and Neutral Camera (INCA) continuously imaged the energetic neutral atoms (ENAs) generated by charge exchange reactions between the energetic, singly ionized trapped magnetospheric ions and the outer atmosphere, or exosphere, of Titan. The images reveal a halo of variable ENA emission about Titan's nearly collisionless outer atmosphere that fades at larger distances as the exospheric density decays exponentially. The altitude of the emissions varies, and they are not symmetrical about the moon, reflecting the complexity of the interactions between Titan's upper atmosphere and Saturn's space environment. PMID- 15890875 TI - Titan's magnetic field signature during the first Cassini encounter. AB - The magnetic field signature obtained by Cassini during its first close encounter with Titan on 26 October 2004 is presented and explained in terms of an advanced model. Titan was inside the saturnian magnetosphere. A magnetic field minimum before closest approach marked Cassini's entry into the magnetic ionopause layer. Cassini then left the northern and entered the southern magnetic tail lobe. The magnetic field before and after the encounter was approximately constant for approximately 20 Titan radii, but the field orientation changed exactly at the location of Titan's orbit. No evidence of an internal magnetic field at Titan was detected. PMID- 15890876 TI - Reconstructing the origin of Andaman Islanders. AB - The origin of the Andaman "Negrito" and Nicobar "Mongoloid" populations has been ambiguous. Our analyses of complete mitochondrial DNA sequences from Onges and Great Andaman populations revealed two deeply branching clades that share their most recent common ancestor in founder haplogroup M, with lineages spread among India, Africa, East Asia, New Guinea, and Australia. This distribution suggests that these two clades have likely survived in genetic isolation since the initial settlement of the islands during an out-of-Africa migration by anatomically modern humans. In contrast, Nicobarese sequences illustrate a close genetic relationship with populations from Southeast Asia. PMID- 15890877 TI - Implementation of the semiclassical quantum Fourier transform in a scalable system. AB - We report the implementation of the semiclassical quantum Fourier transform in a system of three beryllium ion qubits (two-level quantum systems) confined in a segmented multizone trap. The quantum Fourier transform is the crucial final step in Shor's algorithm, and it acts on a register of qubits to determine the periodicity of the quantum state's amplitudes. Because only probability amplitudes are required for this task, a more efficient semiclassical version can be used, for which only single-qubit operations conditioned on measurement outcomes are required. We apply the transform to several input states of different periodicities; the results enable the location of peaks corresponding to the original periods. This demonstration incorporates the key elements of a scalable ion-trap architecture, suggesting the future capability of applying the quantum Fourier transform to a large number of qubits as required for a useful quantum factoring algorithm. PMID- 15890878 TI - Picometer-scale electronic control of molecular dynamics inside a single molecule. AB - Tunneling electrons from a low-temperature (5 kelvin) scanning tunneling microscope were used to control, through resonant electronic excitation, the molecular dynamics of an individual biphenyl molecule adsorbed on a silicon(100) surface. Different reversible molecular movements were selectively activated by tuning the electron energy and by selecting precise locations for the excitation inside the molecule. Both the spatial selectivity and energy dependence of the electronic control are supported by spectroscopic measurements with the scanning tunneling microscope. These experiments demonstrate the feasibility of controlling the molecular dynamics of a single molecule through the localization of the electronic excitation inside the molecule. PMID- 15890879 TI - Glacial/interglacial changes in subarctic north pacific stratification. AB - Since the first evidence of low algal productivity during ice ages in the Antarctic Zone of the Southern Ocean was discovered, there has been debate as to whether it was associated with increased polar ocean stratification or with sea ice cover, shortening the productive season. The sediment concentration of biogenic barium at Ocean Drilling Program site 882 indicates low algal productivity during ice ages in the Subarctic North Pacific as well. Site 882 is located southeast of the summer sea-ice extent even during glacial maxima, ruling out sea-ice-driven light limitation and supporting stratification as the explanation, with implications for the glacial cycles of atmospheric carbon dioxide concentration. PMID- 15890880 TI - Two chemically distinct pools of organic nitrogen accumulate in the ocean. AB - The chemical dynamics of marine dissolved organic nitrogen (DON), a reservoir featuring surface accumulations even in areas where nitrogen limits productivity, have yet to be resolved. We exploited differences in the acid lability of amide bonds within high-molecular-weight (HMW) DON to show that vertical DON profiles result in part from the presence of two chemically distinct pools of amide. Half of HMWDON in surface waters is present as N-acetyl amino polysaccharides. In contrast, nearly all deep-sea HMWDON, and therefore, most HMWDON, is present in amides that resist both chemical hydrolysis and biological degradation. PMID- 15890881 TI - Lichen-like symbiosis 600 million years ago. AB - The fossil record of fungi and lichens is scarce. Here we report the discovery of lichen-like fossils, involving filamentous hyphae closely associated with coccoidal cyanobacteria or algae, preserved in marine phosphorite of the Doushantuo Formation (between 551 and 635 million years old) at Weng'an, South China. These fossils indicate that fungi developed symbiotic partnerships with photoautotrophs before the evolution of vascular plants. PMID- 15890882 TI - The structure of a pH-sensing mycobacterial adenylyl cyclase holoenzyme. AB - Class III adenylyl cyclases contain catalytic and regulatory domains, yet structural insight into their interactions is missing. We show that the mycobacterial adenylyl cyclase Rv1264 is rendered a pH sensor by its N-terminal domain. In the structure of the inhibited state, catalytic and regulatory domains share a large interface involving catalytic residues. In the structure of the active state, the two catalytic domains rotate by 55 degrees to form two catalytic sites at their interface. Two alpha helices serve as molecular switches. Mutagenesis is consistent with a regulatory role of the structural transition, and we suggest that the transition is regulated by pH. PMID- 15890883 TI - Structural basis of energy transduction in the transport cycle of MsbA. AB - We used site-directed spin-labeling and electron paramagnetic resonance spectroscopy to characterize the conformational motion that couples energy expenditure to substrate translocation in the multidrug transporter MsbA. In liposomes, ligand-free MsbA samples conformations that depart from the crystal structures, including looser packing and water penetration along the periplasmic side. Adenosine triphosphate (ATP) binding closes the substrate chamber to the cytoplasm while increasing hydration at the periplasmic side, consistent with an alternating access model. Accentuated by ATP hydrolysis, the changes in the chamber dielectric environment and its geometry provide the likely driving force for flipping amphipathic substrates and a potential exit pathway. These results establish the structural dynamic basis of the power stroke in multidrug-resistant ATP-binding cassette (MDR ABC) transporters. PMID- 15890884 TI - Structure of the ABC transporter MsbA in complex with ADP.vanadate and lipopolysaccharide. AB - Select members of the adenosine triphosphate (ATP)-binding cassette (ABC) transporter family couple ATP binding and hydrolysis to substrate efflux and confer multidrug resistance. We have determined the x-ray structure of MsbA in complex with magnesium, adenosine diphosphate, and inorganic vanadate (Mg.ADP.Vi) and the rough-chemotype lipopolysaccharide, Ra LPS. The structure supports a model involving a rigid-body torque of the two transmembrane domains during ATP hydrolysis and suggests a mechanism by which the nucleotide-binding domain communicates with the transmembrane domain. We propose a lipid "flip-flop" mechanism in which the sugar groups are sequestered in the chamber while the hydrophobic tails are dragged through the lipid bilayer. PMID- 15890885 TI - Single, rapid coastal settlement of Asia revealed by analysis of complete mitochondrial genomes. AB - A recent dispersal of modern humans out of Africa is now widely accepted, but the routes taken across Eurasia are still disputed. We show that mitochondrial DNA variation in isolated "relict" populations in southeast Asia supports the view that there was only a single dispersal from Africa, most likely via a southern coastal route, through India and onward into southeast Asia and Australasia. There was an early offshoot, leading ultimately to the settlement of the Near East and Europe, but the main dispersal from India to Australia approximately 65,000 years ago was rapid, most likely taking only a few thousand years. PMID- 15890886 TI - Induction of protein secretory pathway is required for systemic acquired resistance. AB - In plants, systemic acquired resistance (SAR) is established as a result of NPR1 regulated expression of pathogenesis-related (PR) genes. Using gene expression profiling in Arabidopsis, we found that in addition to controlling the expression of PR genes, NPR1 also directly controls the expression of the protein secretory pathway genes. Up-regulation of these genes is essential for SAR, because mutations in some of them diminished the secretion of PR proteins (for example, PR1), resulting in reduced resistance. We provide evidence that NPR1 coordinately regulates these secretion-related genes through a previously undescribed cis element. Activation of this cis-element is controlled by a transcription factor that is translocated into the nucleus upon SAR induction. PMID- 15890887 TI - Freedom and rules: the acquisition and reprogramming of a bird's learned song. AB - Canary song is hierarchically structured: Short stereotyped syllables are repeated to form phrases, which in turn are arranged to form songs. This structure occurs even in the songs of young isolates, which suggests that innate rules govern canary song development. However, juveniles that had never heard normal song imitated abnormal synthetic songs with great accuracy, even when the tutor songs lacked phrasing. As the birds matured, imitated songs were reprogrammed to form typical canary phrasing. Thus, imitation and innate song constraints are separate processes that can be segregated in time: freedom in youth, rules in adulthood. PMID- 15890888 TI - Quantification of cleaved beta2-microglobulin in serum from patients undergoing chronic hemodialysis. AB - BACKGROUND: Patients on chronic hemodialysis are prone to develop amyloid deposits of misfolded beta(2)-microglobulin (beta(2)M) in osteoarticular tissues. beta(2)M with various deletions/truncations and chemical modifications has been found together with structurally intact beta(2)M in extracts of beta(2)M amyloid fibrils. The state of the circulating population of beta(2)M molecules has not been characterized previously with high-resolution methods. METHODS: We used immunoaffinity-liquid chromatography-mass spectrometry analysis of serum samples to examine whether structurally modified beta(2)M is generated in the circulation. In addition, we developed an immunoassay for the quantification of a cleaved beta(2)M variant in biological fluids based on novel monoclonal antibodies and applied this assay to patient and control sera. RESULTS: A specific alteration compatible with the generation of lysine-58-cleaved and truncated beta(2)M (DeltaK58-beta(2)M) was found in the sera of many (20%-40%) dialysis patients but not in control sera or sera from patients with cerebral amyloidosis (Alzheimer disease). Applied to patient sera, specific immunoassays revealed that dialysis, as expected, significantly lowered the total beta(2)M concentration, but the concentrations of DeltaK58-beta(2)M remained unchanged after dialysis. The results also show that patients dialyzed with less biocompatible membranes have higher serum concentrations of cleaved beta(2)M (mean, 8.5, 1.8, and 0.7 mg/L in cuprophane membrane-dialyzed, polysulfone membrane-dialyzed, and control sera, respectively). CONCLUSIONS: This study for the first time demonstrates and assigns the structure of a specific beta(2)M variant in sera from dialysis patients. Because this variant is conformationally unstable in vitro, it may be involved in in vivo amyloidogenesis. PMID- 15890889 TI - Development of a high-throughput automated analyzer using biochip array technology. AB - BACKGROUND: Use of protein array technology over conventional assay methods has advantages that include simultaneous detection of multiple analytes, reduction in sample and reagent volumes, and high output of test results. The susceptibility of ligands to denaturation, however, has impeded production of a stable, reproducible biochip platform, limiting most array assays to manual or, at most, semiautomated processing techniques. Such limitations may be overcome by novel biochip fabrication procedures. METHODS: After selection of a suitable biochip substrate, biochip surfaces were chemically modified and assessed to enable optimization of biochip fabrication procedures for different test panels. The assay procedure was then automated on a dedicated instrument, and assay performance was determined for a panel of cytokine markers. Assay results were then compared with a commercial method for measurement of cytokine markers. RESULTS: Secondary ion mass spectrometry and x-ray photoelectron spectroscopy demonstrated appropriate and reproducible modification of the biochip surface. Contact-angle studies also confirmed generation of hydrophobic surfaces that enabled containment of droplets for fabrication of discrete test regions. Automation of the biochip assays on a dedicated instrument produced excellent cytokine marker performance with intra- and interassay imprecision <10% for most analytes. Comparison studies showed good agreement with other methods (r = 0.95 0.99) for cytokines. CONCLUSION: Performance data from this automated biochip array analyzer provide evidence that it is now possible to produce stable and reproducible biochips for output of more than 2000 test results per hour. PMID- 15890890 TI - The CAG repeat polymorphism in the androgen receptor gene is associated with HDL cholesterol but not with coronary atherosclerosis or myocardial infarction. AB - BACKGROUND: Age-adjusted morbidity and mortality rates from coronary heart disease (CHD) are higher in men than in women. Androgens are suspected to be responsible for the male disadvantage. The genomic effect of androgens is mediated by the androgen receptor (AR), which has a polymorphic CAG repeat in exon 1. The number of repeats is inversely related to the transcriptional activity of the AR on target genes. METHODS: We investigated the association of this CAG repeat polymorphism with CHD and myocardial infarction (MI) in 2 independent case-control studies involving 544 Caucasian men. RESULTS: The number of CAG repeats in the AR gene correlated significantly with HDL-cholesterol (HDL C) in controls (r = 0.21; P = 0.015). This effect was independent of triglycerides, body mass index, alcohol intake, smoking, and age in a multiple regression model (R(2) = 50%). Despite decreased HDL-C, lower CAG repeat numbers were not associated with increased risk for CHD (odds ratio = 0.82; 95% confidence interval, 0.50-1.36; P = 0.44) or MI in carriers of AR genes with lower CAG repeat numbers (odds ratio = 0.72; 95% confidence interval, 0.37-1.39; P = 0.33). CONCLUSIONS: Shorter, more androgenic AR alleles with fewer CAG repeats are associated with lower HDL-C, but not with an increased risk for CHD or MI, which argues against a detrimental androgen effect on cardiovascular risk under physiologic conditions. PMID- 15890891 TI - Population genotyping of hepatitis C virus by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of short DNA fragments. AB - BACKGROUND: Identifying hepatitis C virus (HCV) genotypes has become increasingly important for determining clinical course and the outcome of antiviral therapy. Here we describe the development of restriction fragment mass polymorphism (RFMP) analysis, a novel matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) assay suitable for high-throughput, sensitive, specific genotyping of multiple HCV species. METHODS: The assay is based on PCR amplification and mass measurement of oligonucleotides containing genotype specific motifs in the 5' untranslated region, into which a type IIS restriction endonuclease recognition was introduced by PCR amplification. Enzymatic cleavage of the products led to excision of multiple oligonucleotide fragments representing variable regions whose masses were determined by MALDI-TOF MS. RESULTS: The RFMP assay identified viral genotypes present at concentrations as low as 0.5% and reliably determined their relative abundance. When sera from 318 patients were analyzed, the RFMP assay exhibited 100% concordance with results obtained by clonal sequencing and identified mixed-genotype infections in 22% of the samples, in addition to several subtype variants. CONCLUSIONS: The RFMP assay has practical advantages over existing methods, including better quantitative detection of mixed populations and detection of genotype variants without need for population-based cloning, enabling reliable viral genotyping in laboratories and efficient study of the relationship between viral genotypes and clinical outcome. PMID- 15890892 TI - Distribution of LDL particle size in a population-based sample of children and adolescents and relationship with other cardiovascular risk factors. AB - BACKGROUND: Smaller, denser LDL particles are associated with an increased risk for cardiovascular diseases (CVD). In youths, data on the distribution of LDL particle size and on its association with other CVD risk factors are limited. METHODS: We determined LDL peak particle size by nondenaturing 2%-16% gradient gel electrophoresis in a representative sample of 2249 youths 9, 13, and 16 years of age who participated in a school-based survey conducted in 1999 in the province of Quebec, Canada. Standardized clinical measurements and fasting plasma lipid, glucose, and insulin concentrations were available. RESULTS: The LDL peak particle size distribution was gaussian. The 5th, 50th (median), and 95th percentiles by age and sex were 255.5-258.6, 262.1-263.2, and 268.1-269.5 A, respectively. The prevalence of the small, dense LDL phenotype (LDL peak particle size 50% ex vivo neutralizing activity against both autologous and heterologous primary isolates. Furthermore, the administration of HIV-VLP(A)s by the i.n. immunization route induces a specific CTL activity, although at lower efficiency than the i.p. route. The HIV-VLP(A)s represent an efficient strategy to stimulate both arms of immunity; furthermore, the induction of specific humoral immunity at mucosal sites, which nowadays represent the main port of entry for HIV-1 infection, is of great interest. All these properties, and the possible cross clade in vivo protection, could make these HIV-VLP(A)s a good candidate for a mono- and multicomponent worldwide preventive vaccine approach not restricted to high-priority regions, such as sub-Saharan countries. PMID- 15890946 TI - Human herpesvirus 7 open reading frames U12 and U51 encode functional beta chemokine receptors. AB - Human herpesvirus 7 (HHV-7), which belongs to the betaherpesvirus subfamily and infects mainly CD4+ T cells in vitro, infects children during infancy. HHV-7 contains two genes, U12 and U51, that encode putative homologs of cellular G protein-coupled receptors. To analyze the biological function of the U12 and U51 genes, we cloned these genes and expressed the proteins in cells. U12 and U51 encoded functional calcium-mobilizing receptors for beta-chemokines, which include thymus and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC), EBI1-ligand chemokine (ELC), and secondary lymphoid-tissue chemokine (SLC), but not for other chemokines, suggesting that the chemokine selectivities of the U12 and U51 products were distinct from those of the known mammalian chemokine receptors. ELC and SLC induced migration in Jurkat cells stably expressing U12, but TARC and MDC did not. In contrast, none of these chemokines induced migration in Jurkat cells stably expressing U51. Together, these data indicate that the products of U12 and U51 may play important and different roles in the pathogenesis of HHV-7 through transmembrane signaling. PMID- 15890947 TI - Pseudotypes of vesicular stomatitis virus with CD4 formed by clustering of membrane microdomains during budding. AB - Many plasma membrane components are organized into detergent-resistant membrane microdomains referred to as lipid rafts. However, there is much less information about the organization of membrane components into microdomains outside of lipid rafts. Furthermore, there are few approaches to determine whether different membrane components are colocalized in microdomains as small as lipid rafts. We have previously described a new method of determining the extent of organization of proteins into membrane microdomains by analyzing the distribution of pairwise distances between immunogold particles in immunoelectron micrographs. We used this method to analyze the microdomains involved in the incorporation of the T cell antigen CD4 into the envelope of vesicular stomatitis virus (VSV). In cells infected with a recombinant virus that expresses CD4 from the viral genome, both CD4 and the VSV envelope glycoprotein (G protein) were found in detergent-soluble (nonraft) membrane fractions. However, analysis of the distribution of CD4 and G protein in plasma membranes by immunoelectron microscopy showed that both were organized into membrane microdomains of similar sizes, approximately 100 to 150 nm. In regions of plasma membrane outside of virus budding sites, CD4 and G protein were present in separate membrane microdomains, as shown by double-label immunoelectron microscopy data. However, virus budding occurred from membrane microdomains that contained both G protein and CD4, and extended to approximately 300 nm, indicating that VSV pseudotype formation with CD4 occurs by clustering of G protein- and CD4-containing microdomains. PMID- 15890948 TI - Enterovirus 70 binds to different glycoconjugates containing alpha2,3-linked sialic acid on different cell lines. AB - Enterovirus 70 (EV70), the causative agent of acute hemorrhagic conjunctivitis, exhibits a restricted tropism for conjunctival and corneal cells in vivo but infects a wide spectrum of mammalian cells in culture. Previously, we demonstrated that human CD55 is a receptor for EV70 on HeLa cells but that EV70 also binds to sialic acid-containing receptors on a variety of other human cell lines. Virus recognition of sialic acid attached to underlying glycans by a particular glycosidic linkage may contribute to host range, tissue tropism, and pathogenesis. Therefore, we tested the possibility that EV70 binds to alpha2,3 linked sialic acid, like other viruses associated with ocular infections. Through the use of linkage-specific sialidases, sialyltransferases, and lectins, we show that EV70 recognizes alpha2,3-linked sialic acid on human corneal epithelial cells and on U-937 cells. Virus attachment to both cell lines is CD55 independent and sensitive to benzyl N-acetyl-alpha-D-galactosaminide, an inhibitor of O linked glycosylation. Virus binding to corneal cells, but not U-937 cells, is inhibited by proteinase K, but not by phosphatidylinositol-specific phospholipase C treatment. These results are consistent with the idea that a major EV70 receptor on corneal epithelial cells is an O-glycosylated, non-glycosyl phosphatidylinositol-anchored membrane glycoprotein containing alpha2,3-linked sialic acid, while sialylated receptors on U-937 cells are not proteinaceous. PMID- 15890949 TI - Cinanserin is an inhibitor of the 3C-like proteinase of severe acute respiratory syndrome coronavirus and strongly reduces virus replication in vitro. AB - The 3C-like proteinase (3CLpro) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is one of the most promising targets for anti-SARS-CoV drugs due to its crucial role in the viral life cycle. In this study, a database containing structural information of more than 8,000 existing drugs was virtually screened by a docking approach to identify potential binding molecules of SARS CoV 3CLpro. As a target for screening, both a homology model and the crystallographic structure of the binding pocket of the enzyme were used. Cinanserin (SQ 10,643), a well-characterized serotonin antagonist that has undergone preliminary clinical testing in humans in the 1960s, showed a high score in the screening and was chosen for further experimental evaluation. Binding of both cinanserin and its hydrochloride to bacterially expressed 3CLpro of SARS-CoV and the related human coronavirus 229E (HCoV-229E) was demonstrated by surface plasmon resonance technology. The catalytic activity of both enzymes was inhibited with 50% inhibitory concentration (IC50) values of 5 microM, as tested with a fluorogenic substrate. The antiviral activity of cinanserin was further evaluated in tissue culture assays, namely, a replicon system based on HCoV-229E and quantitative test assays with infectious SARS-CoV and HCoV-229E. All assays revealed a strong inhibition of coronavirus replication at nontoxic drug concentrations. The level of virus RNA and infectious particles was reduced by up to 4 log units, with IC50 values ranging from 19 to 34 microM. These findings demonstrate that the old drug cinanserin is an inhibitor of SARS-CoV replication, acting most likely via inhibition of the 3CL proteinase. PMID- 15890950 TI - Biological and biochemical characteristics of prion strains conserved in persistently infected cell cultures. AB - Abnormal prion protein (PrP(Sc)) plays a central role in the transmission of prion diseases, but the molecular basis of prion strains with distinct biological characteristics remains to be elucidated. We analyzed the characteristics of prion disease by using mice inoculated with the Chandler and Fukuoka-1 strains propagated in a cultured mouse neuronal cell line, GT1-7, which is highly permissive to replication of the infectious agents. Strain-specific biological characteristics, including clinical manifestations, incubation period as related to the infectious unit, and pathological profiles, remained unchanged after passages in the cell cultures. We noted some differences in the biochemical aspects of PrP(Sc) between brain tissues and GT1-7 cells which were unlikely to affect the strain phenotypes. On the other hand, the proteinase K-resistant PrP core fragments derived from Fukuoka-1-infected tissues and cells were slightly larger than those from Chandler-infected versions. Moreover, Fukuoka-1 infection, but not Chandler infection, gave an extra fragment with a low molecular weight, approximately 13 kDa, in both brain tissues and GT1-7 cells. This cell culture model persistently infected with different strains will provide a new insight into the understanding of the molecular basis of prion diversity. PMID- 15890951 TI - Viral expression of CCL2 is sufficient to induce demyelination in RAG1-/- mice infected with a neurotropic coronavirus. AB - Mouse hepatitis virus strain JHM causes a chronic demyelinating disease in susceptible strains of rodents. Demyelination does not develop in infected RAG1-/ (recombination activation gene-deficient) mice but can be induced by several experimental interventions, including adoptive transfer of virus-specific T cells or antibodies. A common feature of demyelination in these models is extensive infiltration of macrophages/microglia into the white matter. The data obtained thus far do not indicate whether macrophage/microglia infiltration, in the absence of T cells or antibody, is sufficient to mediate demyelination. To determine whether the expression of a single macrophage chemoattractant, in the context of virus infection, could initiate the demyelinating process, we engineered a recombinant coronavirus that expressed the chemokine CCL2/monocyte chemoattractant protein-1. CCL2 has been implicated in macrophage infiltration into the central nervous system and is involved in demyelination in many experimental models of demyelination. Extensive macrophage/microglia infiltration and demyelination has developed in RAG1-/- mice infected with this recombinant virus. Thus, these results suggest that the minimal requirement for demyelination is increased expression of a single macrophage-attracting chemokine in the context of an inflammatory milieu, such as that induced by a viral infection. PMID- 15890952 TI - Differences in the fitness of two diverse wild-type human immunodeficiency virus type 1 isolates are related to the efficiency of cell binding and entry. AB - The ability of one primary human immunodeficiency virus type 1 (HIV-1) isolate to outcompete another in primary CD4+ human lymphoid cells appears to be mediated by the efficiency of host cell entry. This study was designed to test the role of entry on fitness of wild-type HIV-1 isolates (e.g., replicative capacity) and to examine the mechanism(s) involved in differential entry efficiency. The gp120 coding regions of two diverse HIV-1 isolates (the more-fit subtype B strain, B5 91US056, and less-fit C strain, C5-97ZA003) were cloned into a neutral HIV-1 backbone by using a recently described yeast cloning technique. The fitness of the primary B5 HIV-1 isolates and its env gene cloned into the NL4-3 laboratory strain had similar fitness, and both were more fit than the C5 primary isolate and its env/NL4-3 chimeric counterpart. Increased fitness of the B5 over C5 virus was mediated by the gp120 coding region of the env gene. An increase in binding/fusion, as well as decreased sensitivity to entry inhibitors (PSC-RANTES and T-20), was observed in cell fusion assays mediated by B5 gp120 compared to C5 gp120. Competitive binding assays using a novel whole virus-cell system indicate that the primary or chimeric B5 had a higher avidity for CD4/CCR5 on host cells than the C5 counterpart. This increased avidity of an HIV-1 isolate for its cell receptors may be a significant factor influencing overall replicative capacity or fitness. PMID- 15890953 TI - Characterization of human immunodeficiency virus Gag-specific gamma interferon expressing cells following protective mucosal immunization with alphavirus replicon particles. AB - A safe, replication-defective viral vector that can induce mucosal and systemic immune responses and confer protection against many infectious pathogens, such as human immunodeficiency virus type 1 (HIV-1), may be an ideal vaccine platform. Accordingly, we have generated and tested alphavirus replicon particles encoding HIV-1 Gag from Sindbis virus (SIN-Gag) and Venezuelan equine encephalitis virus (VEE-Gag), as well as chimeras between the two (VEE/SIN-Gag). Following intramuscular (i.m.), intranasal (i.n.), or intravaginal (IVAG) immunization with VEE/SIN-Gag and an IVAG challenge with vaccinia virus encoding HIV Gag (VV-Gag), a larger number of Gag-specific CD8+ intracellular gamma interferon-expressing cells (iIFNEC) were detected in iliac lymph nodes (ILN), which drain the vaginal/uterine mucosa (VUM), than were observed after immunizations with SIN Gag. Moreover, a single i.n. or IVAG immunization with VEE/SIN-Gag induced a larger number of cells expressing HIV Gag in ILN, and immunizations with VEE/SIN Gag through any route induced better protective responses than immunizations with SIN-Gag. In VUM, a larger percentage of iIFNEC expressed alpha4beta7 or alpha(Ebeta)7 integrin than expressed CD62L integrin. However, in spleens (SP), a larger percentage of iIFNEC expressed alpha4beta7 or CD62L than expressed alpha(Ebeta)7. Moreover, a larger percentage of iIFNEC expressed the chemokine receptor CCR5 in VUM and ILN than in SP. These results demonstrate a better induction of cellular and protective responses following immunizations with VEE/SIN-Gag than that following immunizations with SIN-Gag and also indicate a differential expression of homing and chemokine receptors on iIFNEC in mucosal effector and inductive sites versus systemic lymphoid tissues. PMID- 15890954 TI - Genetic and cell biological characterization of the vaccinia virus A30 and G7 phosphoproteins. AB - The vaccinia virus proteins A30 and G7 are known to play essential roles in early morphogenesis, acting prior to the formation of immature virions. Their repression or inactivation results in the accumulation of large virosomes, detached membrane crescents, and empty immature virions. We have undertaken further study of these proteins to place them within the context of the F10 kinase, the A14 membrane protein, and the H5 phosphoprotein, which have been the focus of previous studies within our laboratory. Here we confirm that both A30 and G7 undergo F10 kinase-dependent phosphorylation in vivo and recapitulate that modification of A30 in vitro. Although the detached crescents observed upon loss of A30 or G7 echo those seen upon repression of A14, no interaction between A30/G7 and A14 could be detected. We did, however, determine that the A30 and G7 proteins are unstable during nonpermissive tsH5 infections, suggesting that the loss of A30/G7 is the underlying cause for the formation of lacy or curdled virosomes. We also determined that the temperature-sensitive phenotype of the Cts11 virus is due to mutations in two codons of the G7L gene. Phenotypic analysis of nonpermissive Cts11 infections indicated that these amino acid substitutions compromise G7 function without impairing the stability of either G7 or A30. Utilizing Cts11 in conjunction with a rifampin release assay, we determined that G7 acts at multiple stages of virion morphogenesis that can be distinguished both by ultrastructural analysis and by monitoring the phosphorylation status of several viral proteins that undergo F10-mediated phosphorylation. PMID- 15890955 TI - Inhibition of the herpes simplex virus type 1 DNA polymerase induces hyperphosphorylation of replication protein A and its accumulation at S-phase specific sites of DNA damage during infection. AB - The treatment of mammalian cells with genotoxic substances can trigger DNA damage responses that include the hyperphosphorylation of replication protein A (RPA), a protein that plays key roles in the recognition, signaling, and repair of damaged DNA. We have previously reported that in the presence of a viral polymerase inhibitor, herpes simplex virus type 1 (HSV-1) infection induces the hyperphosphorylation of RPA (D. E. Wilkinson and S. K. Weller, J. Virol. 78:4783 4796, 2004). We initiated the present study to further characterize this genotoxic response to HSV-1 infection. Here we report that infection in the presence of polymerase inhibitors triggers an S-phase-specific response to DNA damage, as demonstrated by induction of the hyperphosphorylation of RPA and its accumulation within viral foci specific to the S phase of the cell cycle. This DNA damage response occurred in the presence of viral polymerase inhibitors and required the HSV-1 polymerase holoenzyme as well as the viral single-stranded-DNA binding protein. Treatment with an inhibitor of the viral helicase-primase did not induce the hyperphosphorylation of RPA or its accumulation in infected cells. Taken together, these results suggest that the S-phase-specific DNA damage response to infection is dependent on the specific inhibition of the polymerase. Finally, RPA hyperphosphorylation was not induced during productive infection, indicating that active viral replication does not trigger this potentially detrimental stress response. PMID- 15890956 TI - Mutational analysis of bovine leukemia virus Rex: identification of a dominant negative inhibitor. AB - The Rex proteins of the delta-retroviruses act to facilitate the export of intron containing viral RNAs. The Rex of bovine leukemia virus (BLV) is poorly characterized. To gain a better understanding of BLV Rex, we generated a reporter assay to measure BLV Rex function and used it to screen a series of point and deletion mutations. Using this approach, we were able to identify the nuclear export signal of BLV Rex. Further, we identified a dominant-negative form of BLV Rex. Protein localization analysis revealed that wild-type BLV Rex had a punctate nuclear localization and was associated with nuclear pores. In contrast, the dominant-negative BLV Rex mutation had a diffuse nuclear localization and no nuclear pore association. Overexpression of the dominant-negative BLV Rex altered the localization of the wild-type protein. This dominant-negative derivative of BLV Rex could be a useful tool to test the concept of intracellular immunization against viral infection in a large animal model. PMID- 15890957 TI - Elimination of ie1 significantly attenuates murine cytomegalovirus virulence but does not alter replicative capacity in cell culture. AB - The major immediate-early (MIE) genes of cytomegaloviruses (CMV) are broadly thought to be decisive regulators of lytic replication and reactivation from latency. To directly assess the role of the MIE protein IE1 during the infection of murine CMV (MCMV), we constructed an MCMV with exon 4 of the ie1 gene deleted. We found that, independent of the multiplicity of infection, the resulting recombinant virus, MCMVdie1, which fails to express the IE1 protein, was fully competent for early gene expression and replicated in different cultured cell types with identical kinetics to those of parental or revertant virus. Immunofluorescence microscopy studies revealed that MCMVdie1 was greatly impaired in its capacity to disrupt promyelocytic leukemia bodies in NIH 3T3 cells early after infection, a process that has been proposed to increase viral transcription efficiency. We examined MCMVdie1 in the murine model using both immunocompetent BALB/c and severe combined immunodeficient (SCID) mice. When MCMVdie1 was inoculated into these two types of mice, significantly lower viral titers were detected in infected organs than in those of the wild-type virus-infected animals. Moreover, the ie1-deficient MCMV exhibited a markedly reduced virulence. While all animals infected with 5 x 10(4) PFU of parental virus died by 30 days postinfection, SCID mice infected with a similar dose of MCMVdie1 did not succumb before 60 days postinfection. The in vivo defective growth phenotype of MCMVdie1 was abrogated upon rescue of ie1. These results demonstrate the significance of the ie1 gene for promoting an acute MCMV infection and virulence yet indicate that MCMV is able to grow in vivo, although impaired, in the absence of the ie1 gene. PMID- 15890958 TI - Identification and characterization of the putative fusion peptide of the severe acute respiratory syndrome-associated coronavirus spike protein. AB - Severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is a newly identified member of the family Coronaviridae and poses a serious public health threat. Recent studies indicated that the SARS-CoV viral spike glycoprotein is a class I viral fusion protein. A fusion peptide present at the N-terminal region of class I viral fusion proteins is believed to initiate viral and cell membrane interactions and subsequent fusion. Although the SARS-CoV fusion protein heptad repeats have been well characterized, the fusion peptide has yet to be identified. Based on the conserved features of known viral fusion peptides and using Wimley and White interfacial hydrophobicity plots, we have identified two putative fusion peptides (SARS(WW-I) and SARS(WW-II)) at the N terminus of the SARS-CoV S2 subunit. Both peptides are hydrophobic and rich in alanine, glycine, and/or phenylalanine residues and contain a canonical fusion tripeptide along with a central proline residue. Only the SARS(WW-I) peptide strongly partitioned into the membranes of large unilamellar vesicles (LUV), adopting a beta-sheet structure. Likewise, only SARS(WW-I) induced the fusion of LUV and caused membrane leakage of vesicle contents at peptide/lipid ratios of 1:50 and 1:100, respectively. The activity of this synthetic peptide appeared to be dependent on its amino acid (aa) sequence, as scrambling the peptide rendered it unable to partition into LUV, assume a defined secondary structure, or induce both fusion and leakage of LUV. Based on the activity of SARS(WW-I), we propose that the hydrophobic stretch of 19 aa corresponding to residues 770 to 788 is a fusion peptide of the SARS-CoV S2 subunit. PMID- 15890959 TI - Poliovirus proteins induce membrane association of GTPase ADP-ribosylation factor. AB - Poliovirus infection results in the disintegration of intracellular membrane structures and formation of specific vesicles that serve as sites for replication of viral RNA. The mechanism of membrane rearrangement has not been clearly defined. Replication of poliovirus is sensitive to brefeldin A (BFA), a fungal metabolite known to prevent normal function of the ADP-ribosylation factor (ARF) family of small GTPases. During normal membrane trafficking in uninfected cells, ARFs are involved in vesicle formation from different intracellular sites through interaction with numerous regulatory and coat proteins as well as in regulation of phospholipase D activity and cytoskeleton modifications. We demonstrate here that ARFs 3 and 5, but not ARF6, are translocated to membranes in HeLa cell extracts that are engaged in translation of poliovirus RNA. The accumulation of ARFs on membranes correlates with active replication of poliovirus RNA in vitro, whereas ARF translocation to membranes does not occur in the presence of BFA. ARF translocation can be induced independently by synthesis of poliovirus 3A or 3CD proteins, and we describe mutations that abolished this activity. In infected HeLa cells, an ARF1-enhanced green fluorescent protein fusion redistributes from Golgi stacks to the perinuclear region, where poliovirus RNA replication occurs. Taken together, the data suggest an involvement of ARF in poliovirus RNA replication. PMID- 15890960 TI - Aureusvirus P14 is an efficient RNA silencing suppressor that binds double stranded RNAs without size specificity. AB - RNA silencing is a conserved eukaryotic gene regulatory system in which sequence specificity is determined by small RNAs. Plant RNA silencing also acts as an antiviral mechanism; therefore, viral infection requires expression of a silencing suppressor. The mechanism and the evolution of silencing suppression are still poorly understood. Tombusvirus open reading frame (ORF) 5-encoded P19 is a size-selective double-stranded RNA (dsRNA) binding protein that suppresses silencing by sequestering double-stranded small interfering RNAs (siRNAs), the specificity determinant of the antiviral silencing system. To better understand the evolution of silencing suppression, we characterized the suppressor of the type member of Aureusviruses, the closest relatives of the genus Tombusvirus. We show that the Pothos latent virus (PoLV) ORF 5-encoded P14 is an efficient suppressor of both virus- and transgene-induced silencing. Findings that in vitro P14 binds dsRNAs and double-stranded siRNAs without obvious size selection suggest that P14, unlike P19, can suppress silencing by sequestering both long dsRNA and double-stranded siRNA components of the silencing machinery. Indeed, P14 prevents the accumulation of hairpin transcript-derived siRNAs, indicating that P14 inhibits inverted repeat-induced silencing by binding the long dsRNA precursors of siRNAs. However, viral siRNAs accumulate to high levels in PoLV infected plants; therefore, P14 might inhibit virus-induced silencing by sequestering double-stranded siRNAs. Finally, sequence analyses suggest that P14 and P19 suppressors diverged from an ancient dsRNA binding suppressor that evolved as a nested protein within the common ancestor of aureusvirus-tombusvirus movement proteins. PMID- 15890961 TI - Viral class 1 RNase III involved in suppression of RNA silencing. AB - Double-stranded RNA (dsRNA)-specific endonucleases belonging to RNase III classes 3 and 2 process dsRNA precursors to small interfering RNA (siRNA) or microRNA, respectively, thereby initiating and amplifying RNA silencing-based antiviral defense and gene regulation in eukaryotic cells. However, we now provide evidence that a class 1 RNase III is involved in suppression of RNA silencing. The single stranded RNA genome of sweet potato chlorotic stunt virus (SPCSV) encodes an RNase III (RNase3) homologous to putative class 1 RNase IIIs of unknown function in rice and Arabidopsis. We show that RNase3 has dsRNA-specific endonuclease activity that enhances the RNA-silencing suppression activity of another protein (p22) encoded by SPCSV. RNase3 and p22 coexpression reduced siRNA accumulation more efficiently than p22 alone in Nicotiana benthamiana leaves expressing a strong silencing inducer (i.e., dsRNA). RNase3 did not cause intracellular silencing suppression or reduce accumulation of siRNA in the absence of p22 or enhance silencing suppression activity of a protein encoded by a heterologous virus. No other known RNA virus encodes an RNase III or uses two independent proteins cooperatively for RNA silencing suppression. PMID- 15890962 TI - Loss of interferon regulatory factor 3 in cells infected with classical swine fever virus involves the N-terminal protease, Npro. AB - We show that cells infected with the pestivirus classical swine fever virus (CSFV) fail to produce alpha/beta interferon not only following treatment with double-stranded RNA but also after superinfection with a heterologous virus, the alphavirus Sindbis virus, a virus shown to normally induce interferon. We investigated whether the inhibition of interferon synthesis by CSFV involved a block in interferon regulatory factor 3 (IRF3) activity. Cells infected with CSFV exhibited a lack of translocation of green fluorescent protein-IRF3 to the nucleus; however, constitutive shuttling of IRF3 was not blocked, since it could still accumulate in the nucleus in the presence of leptomycin B. Interestingly subcellular fractionation analysis showed that IRF3 was lost from the cytoplasm of infected cells from 18 h postinfection onwards. Using IRF3 promoter-luciferase reporter constructs, we demonstrate that loss of IRF3 was due to an inhibition of transcription of the IRF3 gene in CSFV-infected cells. Further, we investigated which viral protein may be responsible for the inhibition of interferon and loss of IRF3. We used cell lines expressing the CSFV N-terminal protease (Npro) to show that this single viral protein, unique to pestiviruses, inhibited interferon production in response to Sindbis virus. In addition to being lost from CSFV infected cells, IRF3 was lost from Npro-expressing cells. The results demonstrate a novel viral evasion of innate host defenses, where interferon synthesis is prevented by inhibiting transcription of IRF3 in CSFV-infected cells. PMID- 15890963 TI - Pseudorabies virus glycoprotein gD contains a functional endocytosis motif that acts in concert with an endocytosis motif in gB to drive internalization of antibody-antigen complexes from the surface of infected monocytes. AB - Viral glycoproteins gB and gD of the swine alphaherpesvirus pseudorabies virus (PRV), which is closely related to human herpes simplex virus and varicella zoster virus, are able to drive internalization of antibody-antigen complexes that may form at the cell surface of infected monocytes, thereby protecting these cells from efficient antibody-mediated lysis. We found earlier that gB relies on an endocytosis motif in its cytoplasmic domain for its function during this internalization process. Here, we report that the PRV gD protein also contains a functional endocytosis motif (YRLL) in its cytoplasmic domain that drives spontaneous endocytosis of gD from the cell surface early in infection and that acts in concert with the endocytosis motif in gB to contribute to efficient internalization of antibody-antigen complexes in PRV-infected monocytes. PMID- 15890964 TI - Heat shock protein 90 expression in Epstein-Barr virus-infected B cells promotes gammadelta T-cell proliferation in vitro. AB - The aim of this study was to elucidate the in vitro response of gammadelta T cells to Epstein-Barr virus (EBV)-infected B cells and to determine whether EBV induced heat shock proteins (HSPs) might serve as gammadelta T-cell stimulants. Cytofluorometric analysis revealed HSP90 cell surface expression in 12% of the EBV-immortalized B-cell population in all four of the B-cell lines tested. HSP27, HSP60, and HSP70 were not detected on the cell surface by cytofluorometry in these same B-cell lines. HSP90 and HSP60, but not HSP70 or HSP27, were detected on the cell surface after 125I cell surface labeling and immunoprecipitation with anti-human HSP monoclonal antibodies. In vitro kinetic studies indicated that gammadelta T cells increased at least twofold by day 11 postinfection in cultures of EBV-seronegative peripheral blood lymphocytes infected with EBV, whereas percentages of alphabeta T cells in these same cultures either decreased slightly or remained relatively unchanged in response to EBV infection. Addition of anti human HSP90 monoclonal antibody to the EBV-infected lymphocyte cultures inhibited gammadelta T-cell expansion by 92%. The inhibition of gammadelta T-cell expansion by anti-HSP90 antibody was reversed upon treatment with exogenous HSP90. Taken together, these results indicate that HSP90 played an important role in the stimulation of gammadelta T cells during EBV infection of B cells in vitro and may serve as an important immunomodulator of gammadelta T cells during acute EBV infection. PMID- 15890965 TI - Genetic and biochemical evidence for an oligomeric structure of the functional L polymerase of the prototypic arenavirus lymphocytic choriomeningitis virus. AB - The arenavirus L protein has the characteristic sequence motifs conserved among the RNA-dependent RNA polymerase L proteins of negative-strand (NS) RNA viruses. Studies based on the use of reverse-genetics approaches have provided direct experimental evidence of the key role played by the arenavirus L protein in viral RNA synthesis. Sequence alignment shows six conserved domains among L proteins of NS RNA viruses. The proposed polymerase module of L is located within its domain III, which contains highly conserved amino acids within motifs designated A and C. We have examined the role of these conserved residues in the polymerase activity of the L protein of the prototypic arenavirus, lymphocytic choriomeningitis virus (LCMV), in vivo using a minigenome rescue assay. We show here that the presence of sequence SDD, a characteristic of motif C of segmented NS RNA viruses, as well as the presence of the highly conserved D residue within motif A of L proteins, is strictly required for the polymerase activity of the LCMV L protein. The strong dominant negative phenotype associated with many of the mutants examined and results from coimmunoprecipitation studies provided genetic and biochemical evidence, respectively, for the requirement of the L-L interaction for the polymerase activity of the LCMV L protein. PMID- 15890966 TI - Toll-like receptor signaling inhibits hepatitis B virus replication in vivo. AB - Toll-like receptors (TLR) play a key role in innate immunity. To examine the ability of diverse TLRs to modulate hepatitis B virus (HBV) replication, HBV transgenic mice received a single intravenous injection of ligands specific for TLR2, TLR3, TLR4, TLR5, TLR7, and TLR9. All of the ligands except for TLR2 inhibited HBV replication in the liver noncytopathically within 24 h in a alpha/beta interferon-dependent manner. The ability of these TLR ligands to induce antiviral cytokines at the site of HBV replication suggests that TLR activation could represent a powerful and novel therapeutic strategy for the treatment of chronic HBV infection. PMID- 15890967 TI - Genome replication and progeny virion production of herpes simplex virus type 1 mutants with temperature-sensitive lesions in the origin-binding protein. AB - Genome replication of herpes simplex viruses (HSV) in cultured cells is thought to be started by the action of the virus-encoded origin-binding protein (OBP). In experiments using two HSV-1 mutants with temperature-sensitive lesions in the helicase domain of OBP, we demonstrated that this function is essential during the first 6 hours of the lytic cycle. Once DNA synthesis has started, this function is no longer required, suggesting that origin-driven initiation of viral DNA replication is a single event rather than a continuous process. PMID- 15890968 TI - PECAM-1 mediates NO-dependent dilation of arterioles to high temporal gradients of shear stress. AB - OBJECTIVE: In response to changes in wall shear stress (WSS) the vascular endothelium releases several factors, among others nitric oxide. On the basis of studies of endothelial cells in culture, suggesting that platelet endothelial cell adhesion molecule-1 (PECAM-1) is specifically involved in sensing and coupling high temporal gradients of fluid shear stress with activation of eNOS, we hypothesized that dilations of isolated skeletal muscle arterioles from PECAM 1 knockout mice (PECAM-KO) will be reduced to rapid increases in WSS elicited by increases in perfusate flow. METHODS AND RESULTS: Small and large step increases in flow resulted in substantial dilations in arterioles of WT mice (45+/-4%), but they were markedly reduced in arterioles of PECAM-KO mice (22+/-5%). The initial slope of dilations, when WSS increased rapidly, was greater in vessels of WT than those of PECAM-KO mice (slopes: 0.378 and 0.094, respectively), whereas the second phase of dilations, when flow/shear stress was steady, was similar in the 2 groups (slopes: 0.085 and 0.094, respectively). Inhibition of eNOS significantly reduced the initial phase of dilations in arterioles from WT, but not from those of PECAM-KO mice. The calcium ionophore A23187 elicited similar NO mediated dilation in both WT and PECAM-KO mice. CONCLUSIONS: In isolated arterioles of PECAM-KO mice activation of eNOS and consequent dilation by agonists is maintained, but the dilation to high temporal gradients of wall shear stress elicited by increases in perfusate flow is reduced. Thus, we propose that PECAM-1 plays an important role in the ability of the endothelium to sense and couple high temporal gradients of wall shear stress to NO-mediated arteriolar dilation during sudden changes in blood flow in vivo. PMID- 15890969 TI - Platelet microparticles: a transcellular delivery system for RANTES promoting monocyte recruitment on endothelium. AB - OBJECTIVE: Platelet activation mediates multiple cellular responses, including secretion of chemokines such as RANTES (CCL5), and formation of platelet microparticles (PMPs). We studied the role of PMPs in delivering RANTES and promoting monocyte recruitment. METHODS AND RESULTS: Here we show that PMPs contain substantial amounts of RANTES and deposit RANTES on activated endothelium or murine atherosclerotic carotid arteries. RANTES deposition is facilitated by flow conditions and more efficient than that conferred by PMP supernatants. Interactions of PMPs with activated endothelium in flow were mostly characterized by rolling. RANTES deposition showed a diffuse distribution pattern and was rarely colocalized with firmly adherent PMPs, substantiating that RANTES deposition occurs during transient interactions. Importantly, preperfusion with PMPs enhanced monocyte arrest on activated endothelium or atherosclerotic carotid arteries, which could be inhibited by a blocking antibody or a RANTES receptor antagonist. Blockade or deficiency of PMP-expressed adhesion receptors demonstrated differential requirement of P-selectin, glycoprotein Ib (GPIb), GPIIb/IIIa, and junctional adhesion molecule-A for PMP interactions with endothelium, PMP-dependent RANTES deposition, and subsequent monocyte arrest. CONCLUSION: Circulating PMPs may serve as a finely tuned transcellular delivery system for RANTES, triggering monocyte arrest to inflamed and atherosclerotic endothelium, introducing a novel mechanism for platelet-dependent monocyte recruitment in inflammation and atherosclerosis. PMID- 15890970 TI - Blood oxygen level-dependent MRI of tissue oxygenation: relation to endothelium dependent and endothelium-independent blood flow changes. AB - OBJECTIVES: The contribution of endothelial function to tissue oxygenation is not well understood. Muscle blood oxygen level-dependent MRI (BOLD MRI) provides data largely dependent on hemoglobin (Hb) oxygenation. We used BOLD MRI to assess endothelium-dependent signal intensity (SI) changes. METHODS AND RESULTS: We investigated mean BOLD SI changes in the forearm musculature using a gradient echo technique at 1.5 T in 9 healthy subjects who underwent a protocol of repeated acetylcholine infusions at 2 different doses (16 and 64 microg/min) and N(G)-monomethyl-L-arginine (L-NMMA; 5 mg/min) into the brachial artery. Sodium nitroprusside was used as a control substance. For additional correlation with standard methods, the same protocol was repeated, and forearm blood flow was measured by strain gauge plethysmography. We obtained a significant increase in BOLD SI during acetylcholine infusion (64 microg/min) and a significant decrease for L-NMMA infusion (P<0.005 for both). BOLD SI showed a different kinetic signal than did blood flow, particularly after intermittent ischemia and at high flow rates. CONCLUSIONS: In standard endothelial function tests, BOLD MRI detects a dissociation of tissue Hb oxygenation from blood flow. BOLD MRI may be a useful adjunct in assessing endothelial function. PMID- 15890971 TI - The antiviral cytomegalovirus inducible gene 5/viperin is expressed in atherosclerosis and regulated by proinflammatory agents. AB - OBJECTIVE: Inflammatory processes play an important role in atherosclerosis, and increasing evidence implies that microbial pathogens and proinflammatory cytokines are involved in the development and activation of atherosclerotic lesions. To find new inflammatory genes, we explored the vascular transcriptional response to an activator of innate immunity bacterial lipopolysaccharides (LPSs). METHODS AND RESULTS: Gene arrays identified the cytomegalovirus-inducible gene 5 (cig5)/viperin among the genes most potently induced by LPS in human vascular biopsies. Viperin was expressed by endothelial cells in atherosclerotic arteries and significantly elevated in atherosclerotic compared with normal arteries. In culture, cytomegalovirus infection, interferon-gamma, and LPS induced viperin expression. CONCLUSIONS: Viperin is expressed in atherosclerosis and induced in vascular cells by inflammatory stimuli and cytomegalovirus infection. The putative functions of viperin in atherosclerosis may relate to disease-associated microbes. PMID- 15890972 TI - Breastfeeding and atherosclerosis: intima-media thickness and plaques at 65-year follow-up of the Boyd Orr cohort. AB - OBJECTIVES: The impact of breastfeeding in infancy on cardiovascular disease risk is uncertain. We related breastfeeding in infancy to atherosclerosis in adulthood. METHODS AND RESULTS: A historic cohort study based on a 65-year follow up of the Carnegie (Boyd Orr) survey of diet and health in prewar Britain, 1937 to 1939. A total of 732 eligible cohort members living in or around Aberdeen, Bristol, Dundee, Wisbech, and London were invited for follow-up examinations in 2002, and 405 (55%) participated. In models controlling for age and sex, breastfeeding was inversely associated with common carotid intima-media thickness (IMT; difference -0.03 mm; 95% CI, -0.07 to 0.01), bifurcation IMT (difference 0.19 mm; 95% CI, -0.37 to -0.01), carotid plaque (odds ratio [OR], 0.52; 95% CI, 0.29 to 0.92), and femoral plaque (OR, 0.54; 95% CI, 0.26 to 1.12), compared with bottle-feeding. Controlling for socioeconomic variables in childhood and adulthood, smoking and alcohol made little difference to effect estimates. Controlling for factors potentially on the causal pathway (blood pressure, adiposity, cholesterol, insulin resistance, and C-reactive protein) made little difference to observed associations. CONCLUSIONS: Breastfeeding may be associated with a reduced risk of atherosclerosis in later life. Measurement error and power considerations limit the extent to which conclusions about the mechanisms underlying this relationship can be made. PMID- 15890973 TI - GLUT4 facilitative glucose transporter specifically and differentially contributes to agonist-induced vascular reactivity in mouse aorta. AB - OBJECTIVE: We hypothesized that GLUT4 is a predominant facilitative glucose transporter in vascular smooth muscle cells (VSMCs), and GLUT4 is necessary for agonist-induced VSMC contraction. METHODS AND RESULTS: Glucose deprivation and indinavir, a GLUT4 antagonist, were used to assess the role of GLUT4 and non GLUT4 transporters in vascular reactivity. In isolated endothelium-denuded mouse aorta, approximately 50% of basal glucose uptake was GLUT4-dependent. Norepinephrine-mediated contractions were dependent on both GLUT4 and non-GLUT4 transporters, serotonin (5-HT)-mediated contractions were mainly GLUT4-dependent, and prostaglandin (PG) F(2alpha)-mediated contractions were dependent on non GLUT4 transporters, whereas indinavir had no effect in GLUT4 knockout vessels. We also observed a 46% decrease in GLUT4 expression in aortas from angiotensin II hypertensive mice. Indinavir caused a less profound attenuation of maximal 5-HT mediated contraction in these vessels, corresponding to the lower GLUT4 levels in the hypertensive aortas. Finally, and somewhat surprisingly, chronic GLUT4 knockout was associated with increased vascular reactivity compared with that in wild-type animals, suggesting that chronic absence or reduction of GLUT4 expression in VSMCs leads to opposite effects observed with acute inhibition of GLUT4. CONCLUSIONS: Thus, we conclude that GLUT4 is constitutively expressed in large arteries and likely participates in basal glucose uptake. In addition, GLUT4, as well as other non-GLUT4 facilitative glucose transporters, are necessary for agonist-induced contraction, but each transporter participates in VSMC contraction selectively, depending on the agonist, and changes in GLUT4 expression may account for some of the functional changes associated with vascular diseases like hypertension. PMID- 15890974 TI - Overexpression of VEGF-C causes transient lymphatic hyperplasia but not increased lymphangiogenesis in regenerating skin. AB - Vascular endothelial growth factor (VEGF)-C is necessary for lymphangiogenesis and holds potential for lymphangiogenic therapy in diseases lacking adequate lymphatic drainage. However, the ability of VEGF-C to enhance sustainable, functional lymphatic growth in adult tissues remains unclear. To address this, we evaluated VEGF-C overexpression in adult lymphangiogenesis in regenerating skin. We used a model of mouse tail skin regeneration incorporating a suspension of either VEGF-C overexpressing tumor cells, which provide a continuous supplement of excess VEGF-C to the natural regenerating environment for more than 25 days, or otherwise identical control-transfected tumor cells. We found that excess VEGF C did not enhance the rate of lymphatic endothelial cell (LEC) migration, the density of lymphatic vessels, or the rate of functionality -- even though lymphatic hyperplasia was present early on. Furthermore, the hyperplasia disappeared when VEGF-C levels diminished, which occurred after 25 days, rendering the lymphatics indistinguishable from those in control groups. In vitro, we showed that whereas cell-derived VEGF-C could induce chemoattraction of LECs across a membrane (which involves amoeboid-like transmigration), it did not increase LEC chemoinvasion within a 3-dimensional fibrin matrix (which requires proteolytic migration). These results suggest that whereas excess VEGF-C may enhance early LEC proliferation and cause lymphatic vessel hyperplasia, it does not augment the physiological rate of migration or functionality, and by itself cannot sustain any lasting effects on lymphatic size, density, or organization in regenerating adult skin. PMID- 15890975 TI - Phosphorylation of serine 188 protects RhoA from ubiquitin/proteasome-mediated degradation in vascular smooth muscle cells. AB - cAMP and cyclic GMP-dependent kinases (PKA and PKG) phosphorylate the small G protein RhoA on Ser188. We have previously demonstrated that phosphorylation of Ser188 inhibits RhoA-dependent functions and positively regulates RhoA expression, and that the nitric oxide (NO)/cGMP-dependent protein kinase pathway plays an essential role, both in vitro and in vivo, in the regulation of RhoA protein expression and functions in vascular smooth muscle cells. Here we analyze the consequences of Ser188 phosphorylation on RhoA protein degradation. By expressing Ser188 phosphomimetic wild-type (WT-RhoA-S188E) and active RhoA proteins (Q63L-RhoA-S188E), we show that phosphorylation of Ser188 of RhoA protects RhoA, particularly its active form, from ubiquitin-mediated proteasomal degradation. Coimmunoprecipitation experiments indicate that the resistance of the phosphorylated active form of RhoA to proteasome-mediated degradation is because of its cytoplasmic sequestration through enhanced RhoGDI interaction. In rat aortic smooth muscle cells, stimulation of PKG and inhibition of proteasome by lactacystin, induce nonadditive increases in RhoA protein expression. In addition, stimulation of PKG leads to the accumulation of GTP-bound RhoA in the cytoplasm. In vivo stimulation of the NO/PKG signaling by treating rats with sildenafil increased RhoA level and RhoA phosphorylation, and enhanced its association to RhoGDI in the pulmonary artery, whereas opposite effects are induced by chronic inhibition of NO synthesis in N-omega-nitro-L-arginine-treated rats. Our results thus suggest that Ser188 phosphorylation-mediated protection against degradation is a physiological process regulating the level of endogenous RhoA and define a novel function for RhoGDI, as an inhibitor of Rho protein degradation. PMID- 15890976 TI - Bidirectional ventricular tachycardia and fibrillation elicited in a knock-in mouse model carrier of a mutation in the cardiac ryanodine receptor. AB - Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited disease characterized by adrenergically mediated polymorphic ventricular tachycardia leading to syncope and sudden cardiac death. The autosomal dominant form of CPVT is caused by mutations in the RyR2 gene encoding the cardiac isoform of the ryanodine receptor. In vitro functional characterization of mutant RyR2 channels showed altered behavior on adrenergic stimulation and caffeine administration with enhanced calcium release from the sarcoplasmic reticulum. As of today no experimental evidence is available to demonstrate that RyR2 mutations can reproduce the arrhythmias observed in CPVT patients. We developed a conditional knock-in mouse model carrier of the R4496C mutation, the mouse equivalent to the R4497C mutations identified in CPVT families, to evaluate if the animals would develop a CPVT phenotype and if beta blockers would prevent arrhythmias. Twenty-six mice (12 wild-type (WT) and 14RyR(R4496C)) underwent exercise stress testing followed by epinephrine administration: none of the WT developed ventricular tachycardia (VT) versus 5/14 RyR(R4496C) mice (P=0.02). Twenty-one mice (8 WT, 8 RyR(R4496C), and 5 RyR(R4496C) pretreated with beta blockers) received epinephrine and caffeine: 4/8 (50%) RyR(R4496C) mice but none of the WT developed VT (P=0.02); 4/5 RyR(R4496C) mice pretreated with propranolol developed VT (P=0.56 nonsignificant versus RyR(R4496C) mice). These data provide the first experimental demonstration that the R4496C RyR2 mutation predisposes the murine heart to VT and VF in response caffeine and/or adrenergic stimulation. Furthermore, the results show that analogous to what is observed in patients, beta adrenergic stimulation seems ineffective in preventing life-threatening arrhythmias. PMID- 15890977 TI - Ventricularization of atrial gene expression in the fibrillating heart? PMID- 15890978 TI - A bittersweet modification: O-GlcNAc and cardiac dysfunction. PMID- 15890979 TI - Statin-mediated inhibition of Rho: only to get more NO? PMID- 15890980 TI - Angiogenesis and pericytes in the initiation of ectopic calcification. AB - Ectopic calcification of blood vessels, heart valves, and skeletal muscle is a major clinical problem. There is now good evidence that angiogenesis is associated with ectopic calcification in these tissues and that it is necessary, but not sufficient, for calcification to occur. Angiogenesis may regulate ectopic calcification in several ways. First, many angiogenic factors are now known to exert both direct and indirect effects on bone and cartilage formation. Second, cytokines released by endothelial cells can induce the differentiation of osteoprogenitor cells. Third, the new blood vessels provide oxygen and nutrients to support the growing bone. Finally, the new blood vessels can serve as a conduit for osteoprogenitor cells. These osteoprogenitor cells may be derived from the circulation or from pericytes that are present in the neovessels themselves. Indeed, there is now compelling evidence that pericytes can differentiate into osteoblasts and chondrocytes both in vitro and in vivo. Other vascular cells, including adventitial myofibroblasts, calcifying vascular cells, smooth muscle cells, and valvular interstitial cells, have also been shown to exhibit multilineage potential in vitro. Although these cells share many properties with pericytes, the precise relationship between them is not known. Furthermore, it still remains to be determined whether all or some of these cells contribute to the ectopic calcification observed in vivo. A better understanding of the underlying mechanisms that link angiogenesis, pericytes, and ectopic calcification should provide a basis for development of therapeutic strategies to treat or arrest this clinically significant condition. PMID- 15890981 TI - Adipose tissue, inflammation, and cardiovascular disease. AB - Mounting evidence highlights the role of adipose tissue in the development of a systemic inflammatory state that contributes to obesity-associated vasculopathy and cardiovascular risk. Circulating mediators of inflammation participate in the mechanisms of vascular insult and atheromatous change, and many of these inflammatory proteins are secreted directly from adipocytes and adipose tissue derived macrophages. Several factors linking obesity with an increased cardiovascular risk have been identified. The adipocyte-specific secretory protein adiponectin is a particularly promising candidate in this context. Its levels are decreased in obesity. Adiponectin may mediate some of its demonstrated cardioprotective effects through its anti-inflammatory properties. In addition to decreased expression of beneficial adipokines, secretion of a host of inflammatory factors from visceral adipose tissue may contribute to the increased cardiovascular risk associated with obesity. The cardioprotective effects of many of the most popular drug regimens corroborate these conclusions, demonstrating that along with improvements in other therapeutic end points, they mediate improvements in systemic inflammation. In some cases, these improvements are attributable to direct suppression of inflammatory signaling in adipocytes. The targeted suppression of various proinflammatory cascades in adipocytes specifically represents an exciting new therapeutic opportunity for the cardiovascular disease area. PMID- 15890982 TI - Podosomes at a glance. PMID- 15890983 TI - Talila Volk. Interview by Fiona Watt. PMID- 15890984 TI - IQGAP1: a key regulator of adhesion and migration. AB - The dynamic rearrangement of cell-cell adhesion is one of the major physiological events in tissue development and tumor metastasis. Polarized cell migration, another key event, is a tightly regulated process that occurs during tissue development, chemotaxis and wound healing. Rho-family small GTPases, especially Rac1 and Cdc42, play pivotal roles in these processes through one of their effectors, IQGAP1. Recent studies reveal that IQGAP1 regulates cadherin-mediated cell-cell adhesion both positively and negatively. It captures and stabilizes microtubules through the microtubule-binding protein CLIP-170 near the cell cortex, leading to establishment of polarized cell morphology and directional cell migration. Furthermore, Rac1 and Cdc42 link the adenomatous polyposis coli (APC) protein to actin filaments through IQGAP1 at the leading edge and thereby regulate polarization and directional migration. PMID- 15890985 TI - Inositol-lipid binding motifs: signal integrators through protein-lipid and protein-protein interactions. AB - Inositol lipids have emerged as universal lipid regulators of protein signaling complexes in defined membrane compartments. The number of protein modules that are known to recognise these membrane lipids is rapidly increasing. Pleckstrin homology domains, FYVE domains, PX domains, ENTH domains, CALM domains, PDZ domains, PTB domains and FERM domains are all inositide-recognition modules. The latest additions to this list are members of the clathrin adaptor protein and arrestin families. Initially, inositol lipids were believed to recruit signaling molecules to specific membrane compartments, but many of the domains clearly do not possess high enough affinity to act alone as localisation signals. Another important notion is that some (and probably most) of these protein modules also have protein binding partners, and their protein- and lipid-binding activities might influence one another through allosteric mechanisms. Comparison of the structural features of these domains not only reveals a high degree of conservation of their lipid interaction sites but also highlights their evolutionary link to protein modules known for protein-protein interactions. Protein-protein interactions involving lipid-binding domains could serve as the basis for phosphoinositide-induced conformational regulation of target proteins at biological membranes. Therefore, these modules function as crucially important signal integrators, which explains their involvement in a broad range of regulatory functions in eukaryotic cells. PMID- 15890987 TI - Active finger extension predicts outcomes after constraint-induced movement therapy for individuals with hemiparesis after stroke. AB - BACKGROUND AND PURPOSE: Constraint-induced movement therapy (CIMT) is a rehabilitative strategy used primarily with the post-stroke population to increase the functional use of the neurologically weaker upper extremity through massed practice while restraining the lesser involved upper extremity. Whereas research evidence supports CIMT, limited evidence exists regarding the characteristics of individuals who benefit most from this intervention. The goal of this study was to investigate the potential of 5 measures to predict functional CIMT outcomes. METHODS: A convenience sample of 55 individuals, >6 months after stroke, was recruited that met specific inclusion/exclusion criteria allowing for individuals whose upper extremity was mildly to severely involved. They participated in CIMT 6 hours per day. Pretest, post-test, and follow-up assessments were performed to assess the outcomes for the Wolf Motor Function Test (WMFT). The potential predictors were minimal motor criteria (active extension of the wrist and 3 fingers), active finger extension/grasp release, grip strength, Fugl-Meyer upper extremity motor score, and the Frenchay score. A step-wise regression analysis was used in which the potential predictors were entered in a linear regression model with simultaneous entry of the dependent variables' pretest score as the covariate. Two regressions models were determined for the dependent variable, for immediate post-test, and for follow-up post-test. RESULTS: Finger extension was the only significant predictor of WMFT outcomes. CONCLUSIONS: When using finger extension/grasp release as a predictor in the regression equations, one can predict individual's follow-up scores for CIMT. This experiment provides the most comprehensive investigation of predictors of CIMT outcomes to date. PMID- 15890988 TI - Aggressive therapy with intravenous abciximab and intra-arterial rtPA and additional PTA/stenting improves clinical outcome in acute vertebrobasilar occlusion: combined local fibrinolysis and intravenous abciximab in acute vertebrobasilar stroke treatment (FAST): results of a multicenter study. AB - BACKGROUND AND PURPOSE: A combined therapy of local recombinant tissue plasminogen activator (rtPA) fibrinolysis and intravenous Abciximab platelet inhibition with additional percutaneous transluminal angioplasty (PTA)/stenting may improve recanalization and neurological outcome in patients with acute vertebrobasilar occlusion. METHODS: Combined FAST therapy consisted on intravenous bolus of Abciximab (0.25 mg/kg) followed by a 12-hour infusion therapy (0.125 microg/kg per minute) and low-dose intra-arterial rtPA (median dosage: 20 mg, FAST cohort: N=47). The results were compared with a retrospective cohort, treated by intraarterial rtPA monotherapy (median dosage: 40 mg, rtPA cohort, N=41). Additional PTA/stenting was performed in case of severe residual stenosis. Recanalization success was classified according to the Trials in Myocardial Infarction (TIMI) criteria: TIMI0/1, failed recanalization; TIMI2/3, successful recanalization. Bleeding complications were evaluated according to severe extracerebral hemorrhage (ECH), asymptomatic intracerebral hemorrhage (AIH), and symptomatic intracerebral hemorrhage (SIH). RESULTS: Overall bleeding rate was higher under the combined therapy, but the SIH rate did not differ (FAST versus rtPA: ECH, 3% versus 0%; AIH, 32% versus 22%; SIH 13% versus 12%). Additional PTA/stenting was performed in 14 (FAST) versus 5 (rtPA) patients. TIMI2/3 recanalization rate was similar (FAST, 72%; rtPA, 68%), but TIMI3 rate was remarkably higher under combined therapy (FAST, 45%; rtPA, N=22%). Neurologic outcome appeared better under combined therapy (FAST versus rtPA: favorable outcome rate: 34% versus 17%) with a significantly lower mortality rate (FAST versus rtPA: 38% versus 68%; P=0.006). These results were consistent for embolic and atherothrombotic occlusions. CONCLUSIONS: Combined therapy of intravenous Abciximab and half dose intra-arterial rtPA with additional PTA/stenting appears to improve neurologic outcome in acute vertebrobasilar occlusion despite an increase of overall bleeding complications. PMID- 15890990 TI - Virtual reality-induced cortical reorganization and associated locomotor recovery in chronic stroke: an experimenter-blind randomized study. AB - BACKGROUND AND PURPOSE: Virtual reality (VR) is a new promising computer-assisted technology to promote motor recovery in stroke patients. VR-induced neuroplasticity supporting locomotor recovery is not known. We investigated the effects of VR intervention on cortical reorganization and associated locomotor recovery in stroke patients. METHODS: Ten chronic stroke patients were assigned randomly to either the control group or the VR group. VR was designed to provide interactive real-life practice environments in which practice parameters can be individualized to optimize motor relearning. Laterality index (LI) in the regions of interests (ROIs) and locomotor recovery were measured before and after VR using functional MRI (fMRI) and standardized locomotor tests, respectively. The t test and nonparametric test were performed to compare the mean differences at P<0.05. RESULTS: There was a significant difference in the interval change in the LI score for the primary sensorimotor cortex (SMC) between the groups (P<0.05), indicating that VR practice produced a greater increase in LI for the control group. However, the interval changes in the other ROIs were not significantly different (P>0.05). Motor function was significantly improved after VR (P<0.05). CONCLUSIONS: Our novel findings suggest that VR could induce cortical reorganization from aberrant ipsilateral to contralateral SMC activation. This enhanced cortical reorganization might play an important role in recovery of locomotor function in patients with chronic stroke. This is the first fMRI study in the literature that provides evidence for neuroplasticity and associated locomotor recovery after VR. PMID- 15890989 TI - Acute stroke care in the US: results from 4 pilot prototypes of the Paul Coverdell National Acute Stroke Registry. AB - BACKGROUND AND PURPOSE: The Paul Coverdell National Acute Stroke Registry is being developed to improve the quality of acute stroke care. This article describes key features of acute stroke care from 4 prototype registries in Georgia (Ga), Massachusetts (Mass), Michigan (Mich), and Ohio. METHODS: Each prototype developed its own sampling scheme to obtain a representative sample of hospitals. Acute stroke admissions were identified using prospective (Mass, Mich) or retrospective (Ga, Ohio) methods. All prototypes used a common set of case definitions and data elements. Weighted site-specific frequencies were generated for each outcome. RESULTS: A total of 6867 admissions from 98 hospitals were included; the majority were ischemic strokes (range, 52% to 70%) with transient ischemic attack and intracerebral hemorrhage comprising the bulk of the remainder. Between 19% and 26% of admissions were younger than age 60 years, and between 52% and 58% were female. Black subjects varied from 7.1% (Mich) to 30.6% (Ga). Between 20% and 25% of admissions arrived at the emergency department within 3 hours of onset. Treatment with recombinant tissue plasminogen activator (rtPA) was administered to between 3.0% (Ga) and 8.5% (Mass) of ischemic stroke admissions. Of 118 subjects treated with intravenous rtPA, <20% received it within 60 minutes of arrival. Compliance with secondary prevention practices was poorest for smoking cessation counseling and best for antithrombotics. CONCLUSIONS: A minority of acute stroke patients are treated according to established guidelines. Quality improvement interventions, targeted primarily at the health care systems level, are needed to improve acute stroke care in the United States. PMID- 15890991 TI - Examination of ELN as a candidate gene in the Utah intracranial aneurysm pedigrees. AB - BACKGROUND AND PURPOSE: A study of intracranial aneurysm (IA) sibpairs suggested association of an ELN haplotype with IA risk. Subsequent linkage analysis of the ELN region on chromosome 7q11 in high-risk Utah IA pedigrees significantly confirmed linkage between IA and the ELN region. METHODS: We have investigated the ELN gene as a potential candidate gene for IA in Utah pedigrees. One IA case from each pedigree, who shared an ELN region haplotype segregating in the pedigree, was screened for mutation. The promoter region, 34 exons, and the 3'UTR (UnTranslated Region) of the ELN gene were screened for variants using DHPLC. RESULTS: Variants were observed in the promoter region, exons 4 and 6, and the 3'UTR. Variants in exon 6 and in one 3'UTR position were unique to Utah. The remaining variants were absent in the controls. There was no evidence for segregation of the ELN variants found in IA cases with the hypothesized chromosome 7 haplotypes segregating in pedigrees. CONCLUSIONS: Our analysis does not support ELN as the gene responsible for familial IA in the linked Utah IA pedigrees. PMID- 15890992 TI - The metabolic syndrome is a stronger risk factor for early carotid atherosclerosis in women than in men. AB - BACKGROUND AND PURPOSE: The metabolic syndrome (MetS) is associated with an increased risk for subsequent development of type 2 diabetes mellitus, cardiovascular disease, and stroke. Type 2 diabetes increases the risk of stroke and coronary heart disease in women to a greater extent than in men, and thus the question arises whether there are sex differences in the association of early atherosclerosis and MetS. METHODS: 1588 middle-aged Austrian subjects (1001 males, 587 females) were included in the present study. MetS was defined by the criteria of the National Cholesterol Education Program Adult Treatment Panel III. Early atherosclerosis was assessed by intima-media thickness (IMT) and extent of plaques (B-score) of the carotid arteries. RESULTS: B-score and carotid artery IMT parameters were significantly higher in subjects with the MetS. After adjustment for established risk factors, the difference in B-score remained significant only in women. Computed common carotid artery IMT values using general linear model equations with age, body mass index, and low-density lipoprotein cholesterol as covariates displayed the highest values for men with MetS (811.8+/-9.5 microm). Women with MetS (797.6+/-15 microm) and men without the syndrome (788.8+/-5 microm) showed similar IMTs, whereas women without the MetS presented significantly lower values (735.6+/-7 microm). Among the subcomponents of the MetS, high-density lipoprotein cholesterol showed the strongest impact on IMT in men, whereas blood glucose ranked first in women. CONCLUSIONS: The effect of MetS on early atherosclerosis is more pronounced in females. The impact of the components of MetS on carotid IMT differs between men and women. PMID- 15890993 TI - Use of quantitative magnetic resonance angiography to stratify stroke risk in symptomatic vertebrobasilar disease. AB - BACKGROUND AND PURPOSE: Symptomatic vertebrobasilar disease (VBD) carries a high risk of recurrent stroke. We sought to determine whether a management algorithm consisting of quantitative hemodynamic assessment could stratify stroke risk and guide the need for intervention. METHODS: All patients with symptomatic VBD at our institution are evaluated by a standard protocol including quantitative magnetic resonance angiography (QMRA). Patients are stratified on the basis of the presence or absence of distal flow compromise. Those with low distal flow are offered intervention (surgical or endovascular); all patients receive standard medical therapy. We reviewed the clinical outcome of patients managed with this protocol from 1998 to 2003. RESULTS: Follow-up was available for 47 of 50 patients over a mean interval of 28 months. Stroke and combined stroke/transient ischemic attack free survival at 2 years was calculated using the Kaplan-Meier curve. Patients with normal distal flow (n=31) had an event-free survival of 100% and 96%, respectively. Comparatively, patients with low distal flow (n=16) experienced a 71% and 53% event-free survival, demonstrating a significantly higher risk of recurrent ischemia (P=0.003). Patients with low flow who subsequently underwent treatment (n=12) had an 82% event-free survival. Cox proportional hazards analysis demonstrated that flow status affected event-free survival regardless of covariates. CONCLUSIONS: Patients with symptomatic VBD demonstrating low distal flow on QMRA appear to have a high risk of stroke; conversely, those with normal flow seem to have a benign course and may be optimally managed with medical therapy alone. PMID- 15890994 TI - New concept in cavernous sinus dural arteriovenous fistula: correlation with presenting symptom and venous drainage patterns. AB - BACKGROUND AND PURPOSE: An extradurally located cavernous sinus dural arteriovenous fistula (CSDAVF) exhibits different clinical behavior from other dural arteriovenous fistulas (DAVFs) located between 2 dural leaves. The aim of this study is to define angiographic types of CSDAVF associated with presenting symptom (Sx) and venous drainage patterns. METHODS: CSDAVFs during a mean of 23 month follow-up period of 58 patients (17 to 73 years, male:female ratio=8:50) were retrospectively analyzed. The 3 types of CSDAF, ie, proliferative (PT), restrictive (RT), and late restrictive (LRT) types, were categorized by the degrees and patterns of prominent arteriovenous shunt as well as venous flow. The status of the venous connection with CS and the presenting Sx patterns classified as orbital (OrbSxP), ocular (OcuSxP), cavernous (CavSxP), and cerebral (CerSxP) were associated with angiographic types as well as symptom onset, age, and gender. Correlations of discrete and categorical variables were statistically analyzed using the chi2 or Fisher exact test. RESULTS: PT (n=23) and RT (n=23) of CSDAVF were more common than LRT (n=12) (P=0.016) in patients with younger than 65 years and were related to OrbSxP (P=0.015) and CavSxP (P=0.038) in contrast to LRT to OcuSxP (P=0.004). Early onset of Sxs was related to the OrbSxP (P=0.08) and CavSxP (P<0.001). CerSxP (5%) was noted in RT or LRT. OrbSxP was related to the superior ophthalmic venous drainage (P=0.026) and CavSxP to the inferior petrosal sinus (P=0.046) and posterior fossa venous drainages (P=0.014). Seven patients revealed chronological progression of CSDAVF from PT to LRT and even to complete healing. CONCLUSIONS: CSDAVF presents as 3 distinctive angiographic types and is associated with presenting Sxs and venous drainage patterns. PMID- 15890996 TI - Neuroprotection by hypoxic preconditioning involves oxidative stress-mediated expression of hypoxia-inducible factor and erythropoietin. AB - BACKGROUND AND PURPOSE: Hypoxic preconditioning is an endogenous protection against subsequent lethal hypoxia, but the mechanism involved is not understood. Hypoxia is followed by reactive oxygen species (ROS) production and induces hypoxia-inducible factor (HIF) and its downstream factor erythropoietin (Epo), which is associated with neuroprotection. We hypothesized that these endogenous processes may contribute to hypoxic preconditioning. METHODS: We used a mouse neuronal culture model, with 2 hours of hypoxia as preconditioning followed by 15 hours of hypoxic insult, and examined the expression of HIF-1alpha, Epo, and their downstream proteins by Western blotting. Copper/zinc-superoxide dismutase (SOD1) transgenic (Tg) mice were used to detect the effect of ROS. Cell survival and apoptosis were detected by mitogen-activated protein 2 quantification, apoptotic-related DNA fragmentation, and caspase-3 fragmentation. Antisense Epo was used to block endogenously produced Epo. RESULTS: Hypoxic preconditioning was protective in wild-type (Wt) neurons but not in neurons obtained from SOD1 Tg mice. In Wt neurons, HIF-1alpha and Epo expression showed a greater increase after hypoxia compared with Tg neurons and reached a higher level with preconditioned hypoxia, followed by pJak2, pStat5, and nuclear factor kappaB (NF kappaB) expression. Antisense Epo decreased these downstream proteins and the neuroprotection of hypoxic preconditioning. CONCLUSIONS: Hypoxic preconditioning induces ROS, which may downregulate the threshold for production of HIF-1alpha and Epo expression during subsequent lethal hypoxia, thus exerting neuroprotection through the Jak2-Stat5 and NF-kappaB pathways. PMID- 15890998 TI - Gender differences in outcome of conservatively treated patients with asymptomatic high grade carotid stenosis. AB - BACKGROUND AND PURPOSE: Gender differences are currently becoming increasingly recognized as an important prognostic factor in patients with atherosclerotic disease. We investigated gender-related differences in vascular outcome and mortality of asymptomatic patients with high-grade internal carotid artery (ICA) stenosis. METHODS: We enrolled 525 consecutive patients (325 males with a median age of 72 years and 200 females with a median age of 75 years) from a single center registry who were initially treated conservatively with respect to a neurologically asymptomatic > or =70% ICA stenosis. Patients were followed-up for a median of 38 months (interquartile range, 18 to 65) for major adverse cardiovascular, cerebral, and peripheral vascular events (MACE: combined end point including myocardial infarction, stroke, [partial] limb amputation, and death), vascular mortality, and all-cause mortality. RESULTS: Cumulative MACE free survival rates in males and females at 1, 3, and 5 years were 83%, 65%, 48% versus 85%, 73%, and 67% (P=0.004), respectively. Adjusted hazard ratios for MACE, vascular mortality, and all-cause mortality for males were 1.96 (P=0.016), 2.48 (P<0.001), and 1.70 (P=0.007) as compared with females, irrespective of age, vascular risk factors, comorbidities, and the individual risk status estimated by the American Society of Anesthesiologists (ASA) score. CONCLUSIONS: Male patients with high-grade carotid artery stenosis are at a considerably higher risk for poor outcome than their female counterparts. In particular, the risk for fatal vascular events is substantially increased in males. PMID- 15890999 TI - Screening for obstructive sleep apnea in stroke patients: a cost-effectiveness analysis. AB - BACKGROUND AND PURPOSE: Obstructive sleep apnea (OSA) is common after acute ischemic stroke and predicts poor stroke recovery, but whether screening for OSA and treatment by continuous positive airway pressure (CPAP) improves neurological outcome is unknown. We used a cost-effectiveness model to estimate the magnitude of benefit that would be necessary to make polysomnography (PSG) and OSA treatment cost-effective in stroke patients. METHODS: A decision tree modeled 2 alternative strategies: PSG followed by 3 months of CPAP for those found to have OSA versus no screening. The primary outcome was the utility gained through OSA screening and treatment in relation to 2 common willingness-to-pay thresholds of $50,000 and $100,000 per quality-adjusted life year (QALY). RESULTS: Screening resulted in an incremental cost-effectiveness ratio of $49,421 per QALY. Screening is cost-effective as long as the treatment of stroke patients with OSA by CPAP improves patient utilities by >0.2 for a willingness-to-pay of $50,000 per QALY and 0.1 for a willingness-to-pay of $100,000 per QALY. CONCLUSIONS: A clinical trial assessing the effectiveness of CPAP in improving stroke outcome is warranted from a cost-effectiveness standpoint. PMID- 15891000 TI - Cognitive consequences of thalamic, basal ganglia, and deep white matter lacunes in brain aging and dementia. AB - BACKGROUND AND PURPOSE: Most previous studies addressed the cognitive impact of lacunar infarcts using radiologic correlations that are known to correlate poorly with neuropathological data. Moreover, absence of systematic bilateral assessment of vascular lesions and masking effects of Alzheimer disease pathology and macrovascular lesions may explain discrepancies among previous reports. To define the relative contribution of silent lacunes to cognitive decline, we performed a detailed analysis of lacunar and microvascular pathology in both cortical and subcortical areas of 72 elderly individuals without significant neurofibrillary tangle pathology or macrovascular lesions. METHODS: Cognitive status was assessed prospectively using the Clinical Dementia Rating (CDR) scale; neuropathological evaluation included Abeta-protein deposition staging and bilateral assessment of microvascular ischemic pathology and lacunes; statistical analysis included multivariate models controlling for age, amyloid deposits, and microvascular pathology. RESULTS: Thalamic and basal ganglia lacunes were negatively associated with CDR scores; cortical microinfarcts, periventricular and diffuse white matter demyelination also significantly affected cognition. In a multivariate model, cortical microinfarcts and thalamic and basal ganglia lacunes explained 22% of CDR variability; amyloid deposits and microvascular pathology explained 12%, and the assessment of thalamic and basal ganglia lacunes added an extra 17%. Deep white matter lacunes were not related to cognitive status in univariate and multivariate models. CONCLUSIONS: In agreement with the recently proposed concept of subcortical ischemic vascular dementia, our autopsy series provides important evidence that gray matter lacunes are independent predictors of cognitive decline in elderly individuals without concomitant dementing processes such as Alzheimer disease. PMID- 15891001 TI - Neuropeptide control of fluid balance in insects. AB - This minireview considers various aspects of the control of hydromineral balance in insects with particular reference to the control of diuresis and natriuresis in mosquitoes, with new information on the diuretic peptides of Anopheles gambiae. PMID- 15891002 TI - Dopaminergic inhibition of reproduction in teleost fishes: ecophysiological and evolutionary implications. AB - In many teleosts, dopamine (DA) exerts direct inhibitory control on gonadotropes, counteracting the stimulatory effect of gonadotropin-releasing hormone (GnRH) on gonadotropin release. This dual control by GnRH and DA has been demonstrated in various adult teleosts and has major implications for aquaculture. Because of its unique life cycle, the European eel has provided a powerful model for demonstrating the key role of DA in the control of puberty. Data from tetrapods suggest that the inhibitory role of DA on reproduction is not restricted to the teleosts. Thus, DA inhibitory control could represent an ancient evolutionary component in the neuroendocrine regulation of reproduction that may have been differentially maintained throughout vertebrate evolution. The intensity of DA inhibition, its main site of action, and its involvement in the control of puberty, seasonal reproduction, ovulation, spermiation, or even sex change may differ among classes of vertebrates, as well as within smaller phylogenetic units such as teleosts or mammals. An inhibitory role for DA has been reported also in some invertebrates, indicating that neuronal DA pathways may have been recruited in various groups of metazoa to participate in the control of reproduction. In addition to the incontestable GnRH neurons, the recruitment of DA neurons for the neuroendocrine control of reproduction provides an additional brain pathway for the integration of various species-specific, internal, and environmental cues. In teleosts, the plasticity of the DA neuroendocrine role may have contributed to their large diversity of biological cycles and to their successful adaptation to various environments. PMID- 15891003 TI - Trends in the evolution of the proenkephalin and prodynorphin genes in gnathostomes. AB - The opioid/orphanin gene family provides a model system for analyzing the outcomes of genome duplication events. Recent studies on the proenkephalin gene provide additional evidence that the organizational plan for this gene has been conserved throughout the extensive radiation of the gnathostome vertebrates. However, an analysis of the amino acid sequence of proenkephalin from the zebrafish, Danio rerio, suggests that novel forms of this opioid precursor may be evolving in teleosts. Analyses of sarcopterygian prodynorphin sequences revealed a proenkephalin signature in prodynorphin. Current studies on the opioid/orphanin gene family point to the duplication events that shaped this family occurring prior to the radiation of the gnathostomes. PMID- 15891004 TI - Silk moth neuropeptide hormones: prothoracicotropic hormone and others. AB - The silkworm, Bombyx mori, is a very useful model species, especially in genetics, biochemistry, physiology, and molecular biology, helping researchers unravel the many mysteries involved in the insect life process. The present review describes our early contributions as chemists to the study of the molting and metamorphosis of B. mori. We also present research by Japanese scientists that contributed to the isolation and characterization of peptide hormones from B. mori. PMID- 15891005 TI - Parathyroid glands in calcium regulation and human disease. AB - In humans and other land-living vertebrates, the parathyroids are known to have an overall regulatory role with action as a thermostat in the systemic calcium homeostasis to ensure tight regulation of serum calcium concentrations and appropriate skeletal mineralization. Parathyroid hormone (PTH) is an 84-amino acid peptide, synthesized and released by parathyroid chief cells in response to hypocalcemia. PTH mobilizes calcium by increasing calcium resorption from bone and by raising calcium reabsorption in the proximal kidney tubule. Treatment with active vitamin D can increase vitamin D receptor expression, inhibit growth of parathyroid tumors, and reduce PTH levels in patients with hyperparathyroidism (HPT). Newly developed vitamin D analogues with reduced calcemic effects and with more pronounced antiproliferative effect may provide new favorable treatment for reversal of HPT. PMID- 15891006 TI - Bioinformatic analysis of peptide precursor proteins. AB - Neuropeptides are among the most important signal molecules in animals. Traditional identification of peptide hormones through peptide purification is a tedious and time-consuming process. With the advent of the genome sequencing projects, putative peptide precursor can be mined from the genome. However, because bioactive peptides are usually quite short in length and because the active core of a peptide is often limited to only a few amino acids, using the BLAST search engine to identify neuropeptide precursors in the genome is difficult and sometimes impossible. To overcome these shortcomings, we subject the entire set of all known Drosophila melanogaster peptide precursor sequences to motif-finding algorithms in search of a motif that is common for all prepropeptides and that could be used in the search for new peptide precursors. PMID- 15891007 TI - Urotensin II: ancient hormone with new functions in vertebrate body fluid regulation. AB - Urotensin II (UII), described in many fish species, is secreted by the caudal neurosecretory system, a unique fish neuroendocrine structure. We have examined UII secretion and its control in euryhaline fish, supporting a proposed role in osmoregulation. However, it is now apparent that UII is present in other vertebrates, including mammals. The 12-amino-acid peptide has been highly conserved and the key cyclic region is common from fish to humans. Our UII radioimmunoassay for flounder, directed to this cyclic region, has shown circulating UII levels in humans and rats comparable with those in fish. In mammals, UII cardiovascular effects vary between species, with vasoconstriction only evident in specific vascular beds. The kidney expresses UII receptors and responds to UII administration by a reduction in glomerular filtration rate, urine flow, and excretion of the major ions. Interestingly, plasma levels of UII are chronically elevated in rat models of hypertension. These observations imply an unforeseen role for this ancient fish hormone in the physiological and perhaps pathophysiological regulation of body fluids in higher vertebrates, including humans. PMID- 15891008 TI - Vitellogenesis in the red crab, Charybdis feriatus: contributions from small vitellogenin transcripts (CfVg) and farnesoic acid stimulation of CfVg expression. AB - During reproductive maturation of the female red crab, Charybdis feriatus, the oocytes rapidly accumulate 110- and 78-kDa major polypeptides. Although the hepatopancreas expresses a high level of vitellogenin (CfVg) mRNA, tissue proteins and secreted proteins of the hepatopancreas consist of only small polypeptides. In addition to the 8.0-kb transcripts, many smaller mRNAs specific to the CfVg gene can be detected. These results suggest that the hepatopancreas also produces smaller CfVg transcripts for small CfVg subunits. Using an RT-PCR cloning approach, a population of the small cDNA clones were isolated. Determining the DNA sequence of these clones revealed that these transcripts were most likely the result of alternative splicing and/or alternative expression of the CfVg gene. In vitro treatment of the hepatopancreas fragments with low levels of farnesoic acid stimulated the expression of CfVg. PMID- 15891009 TI - Identification of 26RFa from frog brain: a novel hypothalamic neuropeptide with orexigenic activity in mammals. AB - In the present study, we report the identification, in the frog brain, of a novel neuropeptide, termed 26RFa, that belongs to the RFamide peptide family. The cDNAs encoding the precursors for 26RFa have been characterized in human and rats. In rats, prepro-26RFa mRNA is expressed exclusively in two hypothalamic nuclei involved in the control of feeding behavior. Intracerebroventricular injection of 26RFa in mice induced a dose-dependent increase in food consumption. Taken together, these data indicate that 26RFa is a novel neuropeptide that may have important biological functions in vertebrates. PMID- 15891010 TI - Combinatorial expression of phenotypes of different cell lineages in the rat and mouse pituitary. AB - As studied by single cell RT-PCR of pituitary hormones, we demonstrated that the pituitaries of rats and mice contain a subpopulation of cells that express two or more hormone phenotypes typically belonging to lineages that are branched separately early during embryonic development, such as glycoprotein hormone alpha subunit (alphaGSU) mRNA + PRL mRNA, alphaGSU mRNA + POMC mRNA, and POMC mRNA + GH or PRL mRNA. GnRH in vitro selectively expands the population of cells coexpressing alphaGSU mRNA + PRL mRNA, and CRH selectively increases the proportion of cells coexpressing alphaGSU mRNA + POMC mRNA. Colocalization of alphaGSU + PRL or alphaGSU + POMC could not be detected by double immunofluorescence. This lineage promiscuity was also observed in the pituitary in vivo. PMID- 15891011 TI - Expansion of the superfamily of G-protein-coupled receptors in chordates. AB - We have used sequence hidden Markov models to identify and classify all G-protein coupled receptors (GPCRs) from 13 eukaryotic genomes. We found that the 5 main families found in mammals are present before the divergence of nematodes. We also found that several classes of GPCRs are not present within the vertebrate lineage, whereas they are present in several other taxa. The superfamily of GPCRs has a very dynamic gene repertoire, as evidenced by the several lineage-specific expansions of various families of GPCRs that we found. Further, the number of GPCRs has gradually increased with increased complexity of the organism. PMID- 15891012 TI - In situ hybridization localization of TRH precursor and TRH receptor mRNAs in the brain and pituitary of Xenopus laevis. AB - We examined the distribution of the mRNAs encoding proTRH and the three TRH receptor subtypes (xTRHR1, xTRHR2, and xTRHR3) in the Xenopus laevis CNS and pituitary. A positive correlation was generally observed between the expression patterns of proTRH and xTRHR mRNAs. xTRHRs were widely expressed in the telencephalon and diencephalon, where two or even three xTRHR mRNAs were often simultaneously observed within the same brain structures. In the pituitary, xTRHR2 was selectively expressed in the distal lobe, and xTRHR3 was found exclusively in the intermediate lobe of white background-adapted animals, indicating that, in amphibians, the effect of TRH on alpha-melanotropin (alpha MSH) secretion from melanotrope cells is mediated through the novel receptor subtype xTRHR3. PMID- 15891013 TI - Adipokinetic hormone and the immune responses of locusts to infection. AB - Injections of Bacillus, or of blastospores from the entomopathogenic fungus, Metarhizium anisopliae, activate the prophenoloxidase (PPO) cascade, and coinjection of adipokinetic hormone-I (AKH) enhances and prolongs these responses. When injected concurrently with an immunizing dose of live bacteria, AKH suppresses the appearance of antimicrobial activity and, after a short delay, increases the growth of bacteria within the hemocoel. Injections of live Escherichia coli or Pseudomonas aeruginosa into locusts fail to activate PPO in the hemolymph, even when coinjected with AKH. The coinjection of bacteria and hormone is rarely lethal to the locust. However, if locusts are injected with AKH when they are infected with Metarhizium, they die more rapidly than if no AKH is administered. PMID- 15891014 TI - siRNA produced by recombinant dicer mediates efficient gene silencing in islet cells. AB - RNA interference (RNAi) is emerging as a powerful and convenient tool for studying gene function and genetic variation. RNAi is mediated by 21- to 23 nucleotide-long, small interfering RNAs (siRNA) produced from larger double stranded RNAs in vivo by the RNase III family enzyme Dicer. To overcome the problems associated with the use of predesigned synthetic siRNA molecules, a novel method utilizing the in vitro activity of recombinant Dicer has been developed recently. In nonislet cells, it has been demonstrated that a pool of siRNA, generated by Dicer from in vitro transcribed dsRNA (d-siRNA), mediates convenient, efficient, and reproducible gene silencing in various cell types. The aim of this study was to evaluate the ability of d-siRNA to silence endogenous gene expression in pancreatic islet cells. We observed that liposomal transfection mediates efficient transport of siRNA in up to 90% of dispersed islet cells and that d-siRNA mediates almost complete and nontoxic silencing of an endogenous mRNA, the messenger coding for the nonreceptor tyrosine kinase c Abl. The approach described here using d-siRNA provides an important tool for elucidating gene function in further studies of pancreatic islets and diabetes pathophysiology. PMID- 15891015 TI - Amphibian pheromones and endocrine control of their secretion. AB - Amphibian sex pheromones of 3 urodele (Cynops pyrrhogaster, C. ensicauda, and Plethodon jordani) and 1 anuran (Litoria splendida) species have been isolated and characterized and found to be either small peptides or larger proteins. Each pheromone secreted by the male acts on conspecific females. Endocrine control of pheromone secretion has been best studied in Cynops. The C. pyrrhogaster pheromone, sodefrin, and the C. ensicauda pheromone, silefrin, are generated from their precursor proteins. The sodefrin and silefrin precursor mRNA levels in the abdominal gland of the cloaca are elevated by prolactin and androgen. An increase in the level of both immunoassayable pheromones caused by treatment with these hormones has also been demonstrated. Receptors for both of these hormones have been localized in the abdominal gland. The discharge of sodefrin into the water is elicited by arginine vasotocin. The responsiveness of the female vomeronasal epithelial cells to sodefrin, as estimated by electro-olfactography, is enhanced markedly by a combination of prolactin and estrogen. Sodefrin elevates intracellular calcium levels in vomeronasal epithelial cells. The population of the sodefrin-responsive cells increases during the breeding period. PMID- 15891016 TI - Signal transduction in Rana melanotrope cells: mechanism of action of neurotensin on secretory and electrical activities. AB - The pars intermedia of the frog (Rana esculenta) pituitary, which is composed of a single population of endocrine cells, the melanotrophs, is a very suitable model to study the mode of action of hypophysiotropic neuropeptides. We have recently characterized neurotensin (NT) in Rana esculenta and found that synthetic frog NT (fNT) stimulates the electrical and secretory activities of melanotrophs. By combining biochemical, pharmacological, microfluorimetric, and electrophysiological approaches, we observed that NT stimulates inositol trisphosphate production that provokes Ca(2+) release from intracellular stores. The resulting increase in cytosolic Ca(2+) concentration ([Ca(2+)](c)) activates the secretion of alpha-melanocyte-stimulating hormone (alpha-MSH), stimulates Ca(2+)-dependent protein kinase (PKC) activity, and provokes a depolarizing chloride efflux. PKC reduces the amplitude, whereas membrane depolarization increases the frequency of L- and N-type Ca(2+) currents underlain by the action potential discharge. The complex regulatory processes exerted by NT on Ca(2+) signaling likely generate discrete variations in the [Ca(2+)](c) at various distances from secretory vesicles, contributing to the fine-tuning of alpha-MSH secretion. PMID- 15891017 TI - Analysis of Rab18 and a new golgin in the secretory pathway. AB - Two new amphibian genes have been isolated and characterized from frog melanotropes, and the level of expression of these genes is related to the secretory status of the cells. Both genes, Rab18 and a novel member of the golgin family of proteins, are ubiquitously expressed in endocrine and nonendocrine tissues, and their corresponding proteins appear to show intracellular distributions associated with discrete vesicular and tubular structures, respectively, suggesting that they may play relevant roles in the regulation of the secretory pathway. PMID- 15891018 TI - Expression of neuropeptide Y family peptides in the brain and gut during stages of the life cycle of a parasitic lamprey (Petromyzon marinus) and a nonparasitic lamprey (Ichthyomyzon gagei). AB - The long-term objective of our research is to show that internal factors may be key to triggering metamorphosis and directing the life history types in lampreys (parasitism versus nonparasitism). Since neuropeptide Y family peptides are key players in the endocrine-mediated feeding and reproductive events in mammals, a role for these peptides in the control of feeding behavior and development can be predicted for lampreys. We have investigated the expression pattern of these peptides in the brain and in the gut during different stages of the life cycle of the parasitic lamprey, Petromyzon marinus, and the nonparasitic lamprey, Ichthyomyzon gagei. We provide a description of the cloning and sequencing of P. marinus and I. gagei cDNA for neuropeptide Y (NPY), peptide tyrosine-tyrosine (PYY), and peptide methionine-tyrosine (PMY). Using sequence-specific primers, the mRNA expression patterns for these peptides in brain and gut of larval (ammocoete) and adult (juvenile, prespawning) lampreys were examined by semiquantitative RT-PCR. The observations extend a potential role of neuropeptide Y family peptides in the modulation of feeding behavior and reproductive maturity in lampreys. PMID- 15891019 TI - Frog corticotropin-releasing hormone (CRH): isolation, molecular cloning, and biological activity. AB - Corticotropin-releasing hormone (CRH) was isolated from the brain of the European green frog, Rana esculenta, by combining HPLC purification with radioimmunoassay (RIA) detection. The amino acid sequence SEEPPISLDLTFHLLREVLEMARAEQIAQQAHSNRKLMDII was identical with the sequence of bullfrog (R. catesbeiana) CRH that was deduced from a cDNA encoding the CRH precursor. Synthetic frog CRH enhanced the release of thyroid-stimulating hormone (TSH) from dispersed bullfrog pituitary cells in a concentration-dependent manner. The TSH-releasing activity of a bullfrog hypothalamic extract was decreased by approximately 45% in the presence of the CRH receptor antagonist, alpha-helical CRH(9-41), suggesting that CRH is one of the main TSH-releasing factors present in the bullfrog hypothalamus. PMID- 15891020 TI - Fluid balance in ruminants: adaptation to external and internal challenges. AB - Ruminants are widespread in hot, arid regions. This demands adaptation to large circadian temperature fluctuations and recurrent periods of food and water shortage. Pregnancy and lactation add to the demands on the adaptive mechanisms due to the greater need for food, water, and electrolytes. The blood volume increases to meet the requirements of the fetoplacental unit and the mammary glands. Unlike urine, the milk cannot be concentrated by antidiuretic hormone (vasopressin). During water deprivation, lactating animals therefore become dehydrated more rapidly than nonlactating animals. Nevertheless, desert-adapted lactating ruminants endure frequent periods of water deprivation without incurring bad health. For the offspring living in hot and dry conditions, it is an advantage that the milk is not concentrated, even if the mother has a high antidiuretic hormone concentration to enable her to concentrate the urine. Since ruminants are prey, they need to drink rapidly when they get access to water. The forestomach allows the animals to store water in the reticulorumen. There is no danger of water intoxication even if they drink to satisfaction in a couple of minutes after having lost as much as 30% of their body weight. PMID- 15891021 TI - Stress hormones in mammals and birds: comparative aspects regarding metabolism, excretion, and noninvasive measurement in fecal samples. AB - A multitude of endocrine mechanisms are involved in coping with challenges. Front line hormones to overcome stressful situations are glucocorticoids (GCs) and catecholamines (CAs). These hormones are usually determined in plasma samples as parameters of adrenal activity and thus of disturbance. GCs (and CAs) are extensively metabolized and excreted afterwards. Therefore, the concentration of GCs (or their metabolites) can be measured in various body fluids or excreta. Above all, fecal samples offer the advantages of easy collection and a feedback free sampling procedure. However, large differences exist among species regarding the route and time course of excretion, as well as the types of metabolites formed. Based on information gained from radiometabolism studies (reviewed in this paper), we recently developed and successfully validated different enzyme immunoassays that enable the noninvasive measurement of groups of cortisol or corticosterone metabolites in animal feces. The determination of these metabolites in fecal samples can be used as a powerful tool to monitor GC production in various species of domestic, wildlife, and laboratory animals. PMID- 15891022 TI - Neuronal, neurohormonal, and autocrine control of Xenopus melanotrope cell activity. AB - Amphibian pituitary melanotropes are used to investigate principles of neuroendocrine translation of neural input into hormonal output. Here, the steps in this translation process are outlined for the melanotrope cell of Xenopus laevis, with attention to external stimuli, neurochemical messengers, receptor dynamics, second-messenger pathways, and control of the melanotrope secretory process. Emphasis is on the pathways that neurochemical messengers follow to reach the melanotrope. The inhibitory messengers, dopamine, gamma-aminobutyric acid, and neuropeptide Y, act on the cells by synaptic input from the suprachiasmatic nucleus, whereas the locus coeruleus and raphe nucleus synaptically stimulate the cells via noradrenaline and serotonin, respectively. Autoexcitatory actions are exerted by acetylcholine, brain-derived neurotrophic factor (BDNF), and the calcium-sensing receptor. At least six messengers released from the pituitary neural lobe stimulate melanotropes in a neurohormonal way: corticotropin-releasing hormone, thyrotropin-releasing hormone, BDNF, urocortin, mesotocin, and vasotocin. They all are produced by the magnocellular nucleus and coexist in various combinations in two types of neurohemal axon terminal. Most of the relevant receptors of the melanotropes have been elucidated. Apparently, the neural lobe has a dominant role in activating melanotrope secretory activity. The intracellular mechanisms translating the various inputs into cellular activities like biosynthesis and secretion constitute the adenylyl cyclase-cAMP pathway and Ca(2+) in the form of periodic changes of the intracellular Ca(2+) concentration, known as Ca(2+) oscillations. It is proposed that the pattern of these oscillations encodes specific regulatory information and that it is set by first messengers that control, for example, via G proteins and cAMP-related events, specific ion channel-mediated events in the membrane of the melanotrope cell. PMID- 15891023 TI - Prolactin: fishy tales of its primary regulator and function. AB - Prolactin (PRL) is an important regulator of multiple biological functions, and the control of PRL expression integrates a wide spectrum of molecules throughout vertebrates. PRL-releasing peptide (PrRP) seems to be an essential stimulator of PRL transcription and secretion in teleost pituitary and peripheral organs. In the amphibious euryhaline mudskipper, the localization of mRNA levels of PrRP and PRL as well as their regulation during acclimation to different environments are closely related. The presence of PrRP-PRL axes in the peripheral organs might suggest an ancient history of this axis prior to the evolution of the hypothalamus-pituitary, and it is possible that the PrRP is an original and primary regulator of PRL. In the euryhaline fishes, the permeability of gut of seawater-acclimated fish is generally greater than that of the freshwater (FW) acclimated fish. The modification in the epithelial cell renewal system may play an important role in regulation of the permeability. PRL induces the cell proliferation during FW acclimation, whereas cortisol stimulates both cell proliferation and apoptosis. Indeed, a large proportion of the various actions of PRL seem to be associated directly or indirectly with cell proliferation and/or apoptosis, which might be a primary function of PRL. PMID- 15891024 TI - Photoperiodic regulation of seasonal breeding in birds. AB - Day length-dependent breeding in birds commonly occurs in spring and summer, but may occur after exposure to complex changes in day length, as for example in transequatorial migrants. More rarely, some photoperiodic birds breed when day lengths are decreasing or are short. The flexibility of avian photoperiodic breeding strategies may reflect modifications to a common reproductive photoperiodic neuroendocrine system. This involves an extraretinal photoreceptor and a biological clock, which generates a circadian rhythm of photoinducibility to measure photoperiodic time. The pineal gland is not essential for the reproductive photoperiodic response. The current model of the avian photoperiodic response has been modified to accommodate short day breeders, by incorporating a role for seasonal changes in prolactin secretion in the termination of breeding. Analysis of the sites of expression of clock genes suggests that the biological clock for reproductive photoperiodic time measurement is in the medial basal hypothalamus. Photoperiodic signal transduction may involve a clock-dependent local conversion of thyroxine to triiodothyronine (T(3)) in the medial basal hypothalamus mediated by increased expression of the gene encoding type 2 iodothyronine deiodinase. This photoinduced increase in T(3) may stimulate the release of gonadotrophin-releasing hormone (GnRH) through thyroid hormone receptors in the median eminence. These may mediate retraction of glial cell end feet ensheathing GnRH nerve terminals abutting onto the hypophysial portal vasculature, allowing GnRH to be released to stimulate gonadotrophin secretion. PMID- 15891025 TI - Neuroendocrine regulation of frog adrenocortical cells by neurotensin. AB - We previously characterized the primary structure of neurotensin (NT) from an extract of the intestine of the frog Rana esculenta. In this study, we provide evidence for the involvement of NT in the neurocrine regulation of the secretory activity of frog adrenocortical cells. Immunohistochemical studies revealed that the adrenal gland of R. esculenta is innervated by a dense network of NT immunoreactive fibers. Graded concentrations of frog NT induced a dose-dependent stimulation of corticosterone and aldosterone secretion by frog adrenocortical explants through activation of two receptors with pEC(50) of 9.8 and 6.9. These data support the view that NT, released by nerve fibers within the frog adrenal gland, acts locally to control corticosteroid secretion. PMID- 15891026 TI - Current status of genetic and endocrine factors in the sex change of protandrous black porgy, Acanthopagrus schlegeli (Teleostean). AB - Black porgy, Acanthopagrus schlegeli Bleeker, a marine protandrous hermaphrodite fish, is functionally male for the first 2 years of life, but begins to sexually change to female after the third year. Testicular tissue and ovarian tissue are separated by connective tissue in the bisexual gonad. This sex pattern provides a unique model to study the mechanism of sex change in fish. The annual profiles of plasma estradiol, vitellogenin, and 11-ketotestosterone concentrations in males were significantly different from those in the 3-year-old females. Oral administration of estradiol stimulated high levels of gonadal aromatase activity, plasma luteinizing hormone (LH) levels, and sex change in the 2-year-old fish. Oral administration with aromatase inhibitors for 1 year further blocked the natural sex change in 3-year-old black porgy and all fish became functional males. Transcripts of estrogen receptor (ER), androgen receptor, and gonadotropin receptors in the ovarian tissue of bisexual gonad were significantly less expressed than those in the bisexual testicular tissue. ER and aromatase transcripts were much higher in the vitellogenic ovary than those in the bisexual ovarian tissue. Plasma LH levels were higher in male fish than sex-changing fish during postspawning and nonspawning season in 2(+)-year-old black porgy. We are also conducting investigations on the role of the genetic factors (Dmrt 1, Sox 9, Sf-1, and Dax-1) in sex development and sex change. An endocrine mechanism of sex change in black porgy is proposed. PMID- 15891027 TI - Characterization of a molt-inhibiting hormone (MIH) receptor in the Y-organ of the kuruma prawn, Marsupenaeus japonicus. AB - To characterize a receptor for molt-inhibiting hormone (MIH) in the kuruma prawn, Marsupenaeus japonicus, we performed a binding assay and chemical cross-linking experiments using radiolabeled recombinant MIH ([(125)I]rMIH). The specific binding of [(125)I]rMIH was found in the membrane fraction of the Y-organ (K(d) = 4.76 x 10(-10) M and B(max) = 5.51 x 10(-12) M). Chemical cross-linking between [(125)I]rMIH and the membrane fraction of the Y-organ revealed that the MIH receptor protein has a molecular size of approximately 70 kDa. PMID- 15891028 TI - The diapause of gypsy moth, Lymantria dispar (L.) (Lepidoptera: Lymantriidae). AB - Lymantria dispar undergoes a prolonged obligatory pharate first instar larval diapause. Little information is available on the endocrine regulation of this diapause. The purpose of the present paper was to obtain information on ecdysteroid titers during the initial, middle, and terminal phases of diapause. We also investigated the effect of exogenous ecdysterone on diapause interruption. Ecdysteroid titers, measured by radioimmunoassay, were higher in diapausing eggs. Furthermore, exogenous ecdysterone did not prevent diapause. Ecdysteroids seem to play a role in the induction and maintenance of the pharate first instar larval diapause, with a drop in the ecdysteroid titer being essential for diapause termination. PMID- 15891029 TI - Interaction of PCBs with thyroid hormone levels and time of hatching in chicken embryos. AB - The effects of polychlorinated biphenyls (PCB 77, PCB 153, and the mixture Aroclor 1242) on circulating and intracellular thyroid hormone (TH) levels were studied during chicken embryonic development. We observed no influences of PCB 153 on TH availability. Aroclor 1242 caused a transient increase in the T(3) level in the cerebellum at day 16. Clear effects were only seen with PCB 77 around the period of hatching: a severely reduced TH peak, which normally coincides with the stage of internal pipping, and a considerable delay in the moment of hatching. PMID- 15891030 TI - Immunoexpression of androgen receptors in the reproductive tract of the stallion. AB - The objective of this study was to visualize androgen receptors (ARs) in the testis, epididymis, and prostate of the stallion by means of immunohistochemistry. Nuclear immunostaining was found in all somatic cells in the testis--Leydig, Sertoli, and peritubular myoid cells; in both types of epithelial cells of the epididymis; and in the secretory cells of the prostate. These results indicate that ARs are distributed throughout the reproductive tract cells of the stallion. PMID- 15891031 TI - The phylogenetic relationship of the glutamate and pheromone G-protein-coupled receptors in different vertebrate species. AB - Searches in genomic databases for human, mouse, zebrafish, and pufferfish genes resulted in the identification of more than 180 protein predictions belonging to the glutamate family of G-protein-coupled receptors (GPCRs). Comparison of data sets from the different species showed that most of the receptor subgroups that form the glutamate family are present in both mammalian and bony fish lineage. This finding indicates that these groups share a phylogenetically ancient origin. The present study also shows that the pheromone-receptor subgroup has undergone independent expansions in three of the four species, leaving the human genome completely deprived of all pheromone receptors. PMID- 15891032 TI - In vivo and in vitro exposures of carp and carp tissues to graded concentrations of endocrine disrupting chemicals. AB - The objective of this study was to correlate morphological and biochemical parameters of reproductive activity in fish exposed to graded concentrations of endocrine disrupting chemicals. Two-year-old carps were exposed for two weeks to graded concentrations of ethynyl-estradiol, tamoxifen, and flutamide. For each chemical, morphological alterations of the gonads and liver were described according to biomarker and sex steroid levels. In parallel, isolated follicular cells (FCs) and testis fragments were incubated in the presence or absence of carp pituitary homogenate and 25-hydroxycholesterol. Media were analyzed for sex steroid level evaluations. PMID- 15891033 TI - Divergent stress coping styles in juvenile brown trout (Salmo trutta). AB - Two distinct stress coping styles, proactive and reactive, have been stated in various animal studies. This divergence in coping has also been indicated in salmonid fish. Here, we test the hypothesis that divergent stress coping styles are identifiable in a sea-ranched brown trout population. To that end, we used a series of tests on individual juvenile brown trout, with each test including a common key aspect of the two different coping styles. Using a clustering method (SAS: PROC FASTCLUS), two groups that clearly differed both in blood chemistry (noradrenalin and adrenalin levels) following confinement and in behavior during hypoxia were identified. PMID- 15891034 TI - Bradykinin receptors in the zebrafish (Danio rerio). AB - Ligand interactions of a zebrafish bradykinin (BK) receptor expressed in vitro were characterized by measuring inositol phosphate accumulation. The ligands were analogues of zebrafish BK. Substitutions of Arg1, Gly4, Ser6, Pro7, Leu8, and Arg9 caused greatly reduced potency and maximum response. The Pro3 to Ala analogue had higher potency, but lower maximum response. These and other differences show that the zebrafish BK receptor has a ligand-interaction profile that is distinct from mammalian B1 and B2 receptors and from the previously characterized BK receptor in trout stomach. The results increase our understanding of the evolution of BK receptors and their ligands. PMID- 15891035 TI - Opioid peptides, CRF, and urocortin in cerebrospinal fluid-contacting neurons in Xenopus laevis. AB - The presence of the opioids, beta-endorphin, met-enkephalin, and endomorphin, and of corticotropin-releasing factor (CRF) and the CRF family member, urocortin (Ucn), is described in cerebrospinal fluid-contacting neurons in the brain of the amphibian, Xenopus laevis. PMID- 15891036 TI - Pre-pro-somatostatin-III may have cortistatin-like functions in fish. AB - This study analyzes daily changes in the expression of somatostatin precursors PSS-I and PSS-III (structurally related to cortistatin) in the goldfish brain. The results indicate that PSS-I expression correlates with the light cycle only in optic tectum-thalamus (OT-Tha). PSS-III expression correlates with the light cycle in telencephalon-preoptic area (Tel-POA) and OT-Tha. In Tel-POA, PSS-III reaches a minimum level at the beginning of the active phase and a maximum level late in this phase. These results suggest that PSS-I in OT-Tha and PSS-III in Tel POA and OT-Tha could be involved in the control of the activity cycles in goldfish. PMID- 15891037 TI - Comparative study of family 2 GPCRs in Fugu rubripes. AB - In this study, members of family 2 GPCRs, one of the largest families of receptors in vertebrates, were isolated and characterized in the genome of the Japanese pufferfish, Fugu rubripes, and compared with the orthologous genes in other vertebrates. Phylogenetic analysis carried out with all vertebrate family 2 GPCR members indicated that CALR/CGRPR and CRF are the most divergent receptor group within this family and that the remaining members appear to originate from a common ancestral gene precursor. PMID- 15891038 TI - Expression of proopiomelanocortin and its cleavage enzyme genes in Rana esculenta and Xenopus laevis gonads. AB - Proopiomelanocortin (POMC) is the precursor protein of different hormones and neuropeptides, and the POMC-derived peptides are produced through proteolytic cleavage. Prohormone convertase PC1 and PC2 are enzymes responsible for the cleavage of the POMC prohormone. The coexpression of POMC, PC1, and PC2 genes was previously described in the brain and the pituitary gland of Rana esculenta and Xenopus laevis, but no data are available for the gonad. The present work demonstrates a gonadal POMC convertase gene expression in Rana esculenta and Xenopus laevis. PMID- 15891039 TI - Fra-1 activity in the frog, Rana esculenta, testis. AB - Using an anti-Fos family member antiserum, we previously described, in the testis of Rana esculenta, the presence of a nuclear 43-kDa protein that we hypothesized to be Fra-1. Using an antiserum against Fra-1, we here report on Fra-1 expression, localization, and putative activity in the R. esculenta testis during the annual reproductive cycle. Western blot analysis confirms that the nuclear 43 kDa protein is Fra-1. Immunocytochemistry demonstrates Fra-1 in peritubular myoid cells (PMC), efferent ducts, and blood vessels. We present, for the first time for a vertebrate, experimental evidence that the expression of Fra-1 in PMC is related to its activity during sperm transport from the tubular compartment to the efferent ducts. PMID- 15891040 TI - Analysis of Xenopus melanotrope cell size and POMC-gene expression. AB - Flow cytometry was used to separate Xenopus melanotrope cells according to their size. The cells were then submitted to real-time RT-PCR to determine the level of POMC-gene expression. The results show a positive correlation between cell size and gene expression for cells from black-background (but not white-background) adapted animals. PMID- 15891041 TI - Are the cannabinoids involved in bony fish reproduction? AB - Following the discovery of two CB1 genes in the fish Fugu rubripes, investigations on the phylogeny of endocannabinoids have indicated that this system is highly conserved. Our study demonstrated that CB1 receptors are expressed in the CNS and gonads of two teleosts, Carassius auratus and Pelvicachromis pulcher, and they show a high percentage of sequence identity with Fugu rubripes CB(1A) and Danio rerio CB1. By means of immunohistochemistry for CB1, sGnRH, and TH, we found a codistribution of these signaling molecules in the basal telencephalon/preoptic area, which are key centers for gonadotropic regulation. We therefore suggest that endocannabinoids are possibly involved in modulating fish reproduction at both the central and peripheral levels. PMID- 15891042 TI - Cloning and expression of guanylin-like peptides in teleost fish. AB - Homologues of the guanylin peptide family are expressed in teleost fish. Using the eel as a model euryhaline species, cDNAs for three peptides, guanylin, uroguanylin, and renoguanylin, were cloned and found to be expressed within both renal and intestinal epithelia. Seawater (SW) acclimation resulted in upregulation of uroguanylin mRNA expression in the intestinal epithelia of both immature "yellow" and sexually mature "silver" eels. SW acclimation also resulted in an increase in guanylin mRNA in silver eel intestine, whereas there was no change in renoguanylin mRNA expression under any condition. No changes in expression were found in the kidney following SW acclimation. PMID- 15891043 TI - Features of the circadian clock in the avian pineal gland. AB - The effects of different environmental factors on the circadian rhythm of melatonin release by chicken pineal glands were analyzed in 5-day experiments in vitro. Periodic, brief 4-h illumination at around midnight resulted in phase inversion in 2 days. Exposure to repeated 41 degrees C temperature elevation at around midnight caused no phase shift in the basic melatonin rhythm. Repeated inversion of the polarity of the magnetic field also affected rhythmic melatonin release. Additionally, expressions of Per1, Per2, Per3, Bmal1, Bmal2, Clock, Cry1, and Cry2 clock genes were simultaneously detected in single chicken pineal glands. PMID- 15891044 TI - Growth hormone increases hsc70/hsp70 expression and protects against apoptosis in whole blood preparations from silver sea bream. AB - Preparations of whole blood, from silver sea bream (Sparus sarba), were used for heat shock protein 70 (hsp70) and apoptosis studies. It was found that the expression of both gene members of the hsp70 family (hsc70 and hsp70) were upregulated during acute heat shock. The transcript abundance of both of these genes was increased when cells were exposed to growth hormone (GH) at concentrations of 10 and 100 ng/mL. It was also found that GH at concentrations of 10 and 100 ng/mL could protect whole blood from camptothecin-induced apoptosis as determined by DNA fragmentation analysis. PMID- 15891045 TI - Upregulation of the somatotropic axis is correlated with increased G6PDH expression in Black Sea bream adapted to iso-osmotic salinity. AB - Black sea bream (Mylio macrocephalus) were adapted to salinities of 0 ppt (freshwater), 6 ppt (hypo-osmotic), 12 ppt (iso-osmotic), 33 ppt (seawater), and 50 ppt (hypersaline) for 1 month. Using RT-PCR assays, the expression of pituitary growth hormone (GH) and hepatic insulin-like growth factor 1 (IGF-1) genes were studied. It was found that the transcripts for both of these genes were highest in fish maintained at iso-osmotic salinity. To correlate the expression of GH and IGF-1 with an index of growth, we also measured the transcript levels of glucose-6-phosphate dehydrogenase (G6PDH) in liver. It was found that this transcript was also elevated in iso-osmotically adapted black sea bream. PMID- 15891046 TI - Seasonal changes in the activity of 11 beta-hydroxysteroid dehydrogenase in toad testes. AB - In mammals, glucocorticoids (GC) are inactivated by the oxidative activity of 11beta-hydroxysteroid dehydrogenase (11beta-HSD). To study that mechanism in the testes of Bufo arenarum, 11beta-HSD activity and plasma corticosterone (B) were determined in toads from pre- (PR), post- (P), and reproductive (R) periods. Toad 11beta-HSD is NAD(+)-dependent and strongly inhibited by glycyrrhetinic acid. V(max) is higher in the P period (1.37 vs. R: 0.13 and PR: 0.26 pmol/min-mg protein), whereas K(m) values (around 200 nM) remain constant. Plasma B concentrations (R: 178; PR: 61; P: 114 nM) are close to K(m) values, suggesting that, under baseline conditions, 11beta-HSD could protect testes against GC action. PMID- 15891047 TI - The advantage of absolute quantification in comparative hormone research as indicated by a newly established real-time RT-PCR: GH, IGF-I, and IGF-II gene expression in the tilapia, Oreochromis niloticus. AB - We have developed a real-time RT-PCR that absolutely quantifies the gene expression of hormones using the standard curve method. The method avoids cloning procedures by using primer extension to create templates containing a T7 promoter gene sequence. It is rapid since neither separate reverse transcriptions nor postamplification steps are necessary, and its low detection level (2 pg/mug total RNA) allows precise absolute quantification. Using the method, we have quantified the gene expression of GH, IGF-I, and IGF-II in the tilapia. PMID- 15891048 TI - Development of the circadian melatonin rhythm and its responsiveness to PACAP in the embryonic chicken pineal gland. AB - The embryonic development of the circadian melatonin (MT) rhythm and the responsiveness of embryonic MT secretion to pituitary adenylate cyclase activating polypeptide (PACAP) were investigated. Using dynamic in vitro bioassays, we showed the following: (1) The circadian clock and/or the intracellular signal transduction pathways connecting the clock to the MT synthesizing apparatus appear between days 16 and 18 (E16-18) of embryonic development. Lack of periodical environmental stimuli can lead to a delay in this maturation. (2) Exposure of the embryonic chicken pineal gland to PACAP induces a transitory increase in MT secretion and (3) a transitory increase in cyclic AMP efflux at or before day E13 in vitro. PMID- 15891049 TI - Further studies of lipid droplets in the bombykol-producing pheromone gland of Bombyx mori. AB - Lipid droplets are abundant in the pheromone-producing cells of B. mori at adult eclosion, followed by daily fluctuations in both their size and number. Their dynamics are related to PBAN-stimulated de novo bombykol production. To elucidate associated events, we performed the following: (1) extraction, purification, and partial characterization of lipid droplet-associated proteins found on their surface since their function could possibly be to transport and/or dock putative lipases that are responsible for the lipolysis of triglycerides in them; (2) separation, purification, and initial analysis of lipids carried by lipophorins and lipid transfer particles originating from pupal and adult hemolymph because of their role in the formation and accumulation of lipid droplets. PMID- 15891050 TI - Androgen receptor in the ovary of postnatal bank vole females. AB - In this experiment, we investigated the influence of the photoperiod upon androgen receptor (AR) distribution and steroid concentrations in ovaries of 21 day-old bank vole females. Sections (6 mum) were assayed immunohistochemically. ARs were localized in the nuclei of granulosa cells. The strongest immunoreaction was observed in primary and early antral follicles, and declined during follicular development. P(4), A, and E(2) contents were measured by RIAs in ovarian homogenates from animals kept in two photoperiods. Ovaries of animals kept in a long photoperiod (18:6 light:dark), which stimulates gonadal activity, contained more P(4) than those kept in a short one (6:18 light:dark). There was no difference in levels of A and E(2). PMID- 15891051 TI - Hormonal control of the IGF system in the sea bream ovary. AB - Numerous studies have described the presence of an intragonadal IGF system involved in regulation of gametogenesis in teleost fish. In the present study, the in vivo effects of estradiol-17beta (E2) and growth hormone (GH) exposure on IGF-I, IGF-II, IGF1R, and IGFBP2 gene expression in sea bream ovary were monitored by RT-PCR during prereproductive and reproductive periods. The evidence demonstrates that both hormones investigated here affect the ovarian IGF system, showing that it is not only under GH control, but also can be regulated by sexual hormones; this hormonal modulation is related to reproductive phase. PMID- 15891052 TI - High species variation within the repertoire of trace amine receptors. AB - Using a variety of search strategies, we obtained the complete or nearly complete repertoire of trace amine receptors from humans, mice, rats, zebrafish, pufferfish, and a number of invertebrates. We found that the number of functional receptors varies from 5 to 50 in each genome, showing that this family of GPCRs has a very dynamic gene repertoire. We show that the previously cloned and characterized GPCRs from insects and mollusks are more closely related to mammalian serotonin, dopamine, and adrenalin receptors than to mammalian TA receptors. This suggests that the ability to bind TAs has arisen independently in different developmental lineages. PMID- 15891053 TI - The use of real-time PCR to study the expression of thyroid hormone receptor beta 2 in the developing chicken. AB - Thyroid hormones and their receptors (TRs) have critical functions in development and metabolism. In chicken, three TRs are known: TRalpha, TRbeta0, and TRbeta2. The latter was isolated from chicken eye, but its presence in other tissues has not yet been extensively investigated. We therefore developed a real-time PCR assay using a Taqman probe and primers based on the unique amino-terminal region of TRbeta2. We detected a strong TRbeta2 mRNA signal in the pituitary, confirmed with in situ hybridization, and in several other tissues. TRbeta2 mRNA was more abundant in the pituitary of newly hatched chicks than in 15-day-old embryos. PMID- 15891054 TI - Neurodistribution of androgen receptor immunoreactivity in the male frog, Rana esculenta. AB - Sexual behavior in vertebrates depends on the cyclic release of steroids and their binding to the brain receptors. Previously, we demonstrated the presence of specific binding of (3)H-testosterone and staining with PG-21 in the brain of the adult male frog, Rana esculenta. Here, we report our further receptor characterization using an anti-androgen receptor antiserum, PG-21, and the androgen site of action in frog brain. Nuclei, which contained cells labeled for the androgen receptor (AR), were mainly identified in the olfactory bulbs, preoptic-septal region, infundibulum, amygdala, thalamus, tectum, torus semicircularis, and medulla. The neuroanatomical AR staining appears similar to that in other lower vertebrates. PMID- 15891055 TI - Pharmacological characterization of melanocortin receptors in fish suggests an important role for ACTH. AB - The melanocortin (MC) receptor subtypes have distinctive characteristic binding profiles. We found that the trout and Fugu MC4 receptors have similar affinity for alpha-MSH and beta-MSH and a much higher affinity for ACTH than does the human MC4 receptor. The Fugu MC1 and the trout and Fugu MC5 receptors also have higher affinity for ACTH-derived peptides than alpha-, beta-, or gamma-MSH. It is tempting to speculate that ACTH-derived peptides may have played an important role as "original" ligands at the MC receptors, while the specificity of the different subtypes for the alpha-, beta-, and gamma-MSH peptides may have appeared at later stages during vertebrate evolution. PMID- 15891056 TI - Measurement of PTHrP, PTHR1, and CaSR expression levels in tissues of sea bream (Sparus aurata) using quantitative PCR. AB - A quantitative PCR (Q-PCR) method has been established to measure the mRNA expression levels of parathyroid hormone-related protein (PTHrP), parathyroid hormone receptor type 1 (PTHR1), and calcium-sensing receptor (CaSR) in sea bream (Sparus aurata), using the housekeeping gene, beta-actin, as endogenous control. TaqMan primers and probes were designed using the Primer Express program, according to the published/unpublished sequences of the three target genes and beta-actin of sea bream. Different tissues including gill, kidney, duodenum, hindgut, rectum, liver, heart, brain, pituitary, skin, muscle, and gonad were removed and immediately snap-frozen from three juvenile sea bream (100-150 g) cultured in sea water. The mRNAs were extracted and reverse-transcribed into cDNAs, which were subsequently examined by the ABI 5700 system using an optimized Q-PCR method. Triplicate measures of each sample indicated consistency of the technique. However, the mRNA expression levels for each transcript in these tissues were variable between fish and also relatively low. Nevertheless, this methodology can be used in the future studies of factors that may alter gene expression in these tissues. PMID- 15891057 TI - Regulation of the stress response in early vertebrates. AB - The acute stress response is a key regulatory system for the maintenance of homeostatic equilibrium that is activated upon an imminent or ongoing disturbance of the "milieu interieur". In general, the stress response in bony fish is similar to that of mammals. The recent cloning and characterization of corticotropin-releasing hormone-binding protein (CRH-BP) in carp (Cyprinus carpio L.) reflects the conservation of the CRH signaling system throughout vertebrates. Now, we can start to investigate the processes that are mediated by the factors that make up this system. The stress response is only one of these processes. PMID- 15891058 TI - Expression of aromatase mRNA in the abdominal gland of the newt, Cynops pyrrhogaster. AB - As a step to see whether the local aromatization mechanism exists in the newt, Cynops pyrrhogaster, cDNA cloning and characterization of newt cytochrome P450 aromatase (P450arom) were conducted. Newt P450arom cDNA was obtained from a C. pyrrhogaster ovarian cDNA library. By RT-PCR and in situ hybridization, an intense P450arom mRNA expression was detected in the abdominal gland. Employing biochemical techniques combined with HPLC and TLC analyses, we also demonstrated the conversion of testosterone to estradiol in the abdominal grand. The significance of aromatization of testosterone in this gland is discussed. PMID- 15891059 TI - Regionally specific occurrence of an active sodefrin variant in the red-bellied newt. AB - Sodefrin (SIPSKDALLK) is a female-attracting pheromone that is secreted by the abdominal gland of the male red-bellied newt. We found that mRNA encoding a sodefrin variant, [Val(8)] sodefrin, is expressed exclusively in specimens captured in the Nara area of Japan. The synthetic peptide was tested for its activity. It attracted females from Nara, but not those from other regions, suggesting that there is a geographic variation in the pheromone molecule and in the responsiveness to the pheromone. Employing an abdominal gland extract and synthetic substrates, the possibility of generation of the putative pheromone, [Val(8)] sodefrin, from the precursor molecule was demonstrated. PMID- 15891060 TI - Androgen metabolism in invertebrates and its modulation by xenoandrogens: a comparative study. AB - Marisa cornuarietis (Mollusc), Hyalella azteca (Crustacean), and Paracentrotus lividus (Echinoderm) demonstrated the ability to metabolize androgens through different pathways catalyzed by 5alpha-reductases (5alpha-R), hydroxysteroid dehydrogenases (HSD), hydroxylases, sulfotransferases (SULT), and fatty-acid acyl CoA acyltransferases (ATAT). Interspecies differences and tissue-specific distribution of those enzymatic activities were observed. Xenobiotics, such as triphenyltin, tributyltin, and fenarimol, interfered with some of the pathways studied, namely, testosterone sulfation, testosterone esterification, and 5alpha R activity. The work evidenced different sensitivity of those pathways to androgenic compounds, together with interphyla differences in androgen metabolism. PMID- 15891061 TI - Expression analysis of growth and energy regulation-associated genes in two divergent chicken strains. AB - We have studied differential expression of genes using cDNA arrays in the hypothalamic region of two divergent chicken lines, which differ in body weight and feeding behavior. Several transcripts from genes in metabolic networks as well as from retroviruses were differentially expressed. PMID- 15891062 TI - The involvement of thyroid hormone metabolism in gilthead sea bream (Sparus auratus) osmoregulation. AB - We have investigated the effect of adaptation to low salinity water on the thyroid status of the euryhaline teleost, Sparus auratus. We show that, following low salinity adaptation, the plasma T(4) concentration increases and branchial deiodination activities of T(4), T(3), and rT(3) decrease. Moreover, branchial and hepatic enzyme activities that are putatively involved in thyroid hormone metabolism respond differentially in low salinity conditions. Our results indicate the involvement of thyroid hormones in Sparus auratus osmoregulation. Moreover, the gills appear well equipped to play an important role in the modulation of plasma thyroid hormone titers. PMID- 15891063 TI - Agouti-related proteins (AGRPs) and agouti-signaling peptide (ASIP) in fish and chicken. AB - We performed an intensive search on sequence databases to identify orthologues of ASIP and AGRP peptides in a number of different species, revealing a number of genomic fragments coding for the C-terminal part of agouti-related motifs, different from annotated peptide sequences, including one fragment from chicken, two from zebrafish, two from Fugu (Takifugu rubripes), and three from Tetraodon (Tetraodon nigroviridis). We have thus shown for the first time that both AGRP and ASIP genes exist in many species in "lower vertebrates" and were most probably present in early stages of vertebrate evolution. PMID- 15891064 TI - The ancestry of the prolactin-releasing hormone precursor. AB - The prolactin-releasing hormone (PRLH) is implicated in food intake and is expressed in several parts of the mammalian brain. The origin of the peptide precursor (PRH) has been unclear, and the only feature resembling other known human neuropeptide sequences is the C-terminal RF-motif, also present in the neuropeptide FF and the neuropeptide RF amide-related peptide families (RFRP). We have recently found sequences of PRH and the closely related precursor C-RF amide in chicken, shedding light on the PRH ancestry. PMID- 15891065 TI - Time-course effects of centrally administered native urotensin-II on motor and cardioventilatory activity in trout. AB - Although in most vertebrate species urotensin-II (UII) is synthesized in neurons of the central nervous system, little is known regarding the physiological actions of UII in the brain. We have investigated the effects of intracerebroventricular (ICV) administration of synthetic trout UII (1, 5, and 50 pmol) on total motor activity (ACT), ventilatory frequency (VF), ventilatory amplitude (VA), and heart rate (HR) in the unanesthetized trout. ICV injection of UII increased ACT in a dose-dependent manner, and the maximal effect was observed at a dose of 5 pmol. At doses of 1 and 5 pmol, UII did not affect VF, VA, or HR. At the highest dose tested (50 pmol), UII not only increased ACT, but also significantly activated VF, VA, and HR. In contrast, ICV injection of synthetic trout angiotensin-II (5 pmol) did not produce any effect on ACT, VF, or VA, but sharply increased HR. These data provide the first evidence that UII can act centrally to induce motor activity in a nonmammalian vertebrate species. PMID- 15891066 TI - Pufferfish and zebrafish have five distinct NPY receptor subtypes, but have lost appetite receptors Y1 and Y5. AB - The two neuropeptide Y (NPY) receptors Y1 and Y5 stimulate feeding in mammals, but are missing in the euteleosts, zebrafish and pufferfish (Takifugu rubripes). Both species have five other subtypes called Y2, Y7, Ya, Yb, and Yc. RT-PCR studies in pufferfish show that all five are expressed in the brain and may mediate NPY effects on feeding. Y2, Ya, and Yb are also broadly expressed in peripheral organs. These results reveal interesting differences in the NPY system of teleosts and mammals that may have arisen in the genetic turmoil involving the basal ray-fin fish tetraploidization. PMID- 15891067 TI - Subtype-specific pharmacological properties of the melanocortin receptors in chicken. AB - The melanocortin (MC) receptors belong to the family of G-protein-coupled receptors and have five subtypes (MC1-5) in mammals and chicken. We have expressed the chicken MC1-5 (cMC1-5) receptors and seven allelic variants of the cMC1 receptor and performed pharmacological characterization of these receptors. Three variants of the cMC1 receptor, all sharing a Glu to Lys mutation in position 92, are associated with dark feather coloration, and these were pharmacologically described as constitutively active. The pharmacological characterization of the cMC1-5 receptors provides insight of how this receptor family has evolved and useful information for the use of chicken in physiological experiments. PMID- 15891068 TI - Environmental agent susceptibility assessment using existing and novel biomarkers as rapid noninvasive testing methods. AB - This study is part of a project aimed at developing and validating novel noninvasive methods for the detection of biomarkers of endocrine disrupters (EDs) directly in the mucus of aquatic species, to identify novel functional biomarker(s) for EDs, and to verify their applicability for field studies. The multidisciplinary approach chosen aims at the development of an integrated testing strategy utilizing in vitro protocols to identify water and sediment fractions with potential endocrine-disrupting activity; the identification, characterization, and measurement of new biomarker(s) for EDs; the development and validation of a dipstick-based test method; and the development of (computer assisted) predictive models. Some results of the first year of the project are presented here. PMID- 15891069 TI - Characterization of PrPc-immunoreactive cells in monkey (Macaca fascicularis) gastrointestinal tract. AB - The gastrointestinal tract (GIT) is one of the most likely entry sites for the pathological isoform of prions (PrP(sc)). To understand how PrP(sc) crosses the digestive mucosa, it is crucial to characterize the cells expressing normal prion protein (PrP(c)). By means of double immunofluorescence applied to sections of the monkey GIT, we demonstrated that, in the stomach, PrP(c) immunostaining occurs in subsets of histamine, somatostatin (Som), ghrelin (Ghr), gastrin (G), and serotonin (5HT) cells. In the small and large bowels, PrP(c) cells were found in subpopulations of cells immunolabeled for 5HT, Som, G, and peptide YY (PYY). PMID- 15891070 TI - Comparative study of PrPc expression in rat, monkey, and cow gastrointestinal tract. AB - The gastrointestinal tract (GIT) appears to be the main site of entry for the pathological isoform of prions (PrP(sc)). To understand how the PrP(sc) internalization process occurs, it is important to characterize the cell types that express normal prion protein (PrP(c)) along the GIT. To do so, we studied the distribution of PrP(c) in the rat, monkey, and cow GIT. Using Western blot analysis, we found that PrP(c) is expressed in all digestive regions of the three species. Immunoreactivity for PrP(c) was found throughout the GIT in epithelial cells sharing the neuroendocrine (NE) phenotype. Immunostained cells appeared scattered throughout the epithelium of fundic and pyloric glands as well as in intestinal villi and crypts. PMID- 15891071 TI - Effect of cortisol on aquaporin expression in the esophagus of the European eel, Anguilla anguilla. AB - Long-term cortisol infusion into freshwater (FW)-adapted eels induced a significant increase in aquaporin-1 (AQP1) mRNA expression within the esophageal epithelium of migratory "silver" eels, but not in nonmigratory, immature "yellow" eels. Cortisol treatment had no significant effect on the mRNA abundance of a second aquaporin-1 isoform, termed AQP1dup, which exhibited a highly variable expression profile among individual members of all fish groups. These results suggest that cortisol, at plasma concentrations similar to that found during FW/seawater (SW) acclimation, induces upregulation in AQP1 expression and thus increases esophageal water permeability during the migration of eels to the SW environment. PMID- 15891072 TI - Developmental expression of NPY/PYY receptors zYb and zYc in zebrafish. AB - This study describes the developmental expression of zYb and zYc receptors in zebrafish. RT-PCR demonstrated that both mRNAs are present from 24-hour postfertilization (hpf) and that their expression increased during larval development. Whole-mount in situ hybridization showed zYb mRNA expression in the epithalamus (24 hpf), telencephalon, hypothalamus, rhombencephalon (24-96 hpf), mesencephalon (48-96 hpf), hatching gland, and otic vesicle (48-64 hpf). zYc mRNA is expressed in the epithalamus (24 and 96 hpf), telencephalon (48-96 hpf), spinal cord, notochord (64 hpf), and hatching gland (96 hpf). These results show that zYb and zYc receptors are expressed during ontogeny, suggesting a role for neuropeptide Y (NPY) and/or peptide YY (PYY) in organogenesis. PMID- 15891073 TI - Successfully determining the sex of adult Varanus mertensi (Reptilia: Varanidae) using a combination of both hemipenile eversion and the ratio of androgens:estradiol in plasma. AB - In remote field localities, given limited access to specialized equipment, determining the sex of captured adult varanid lizards before release can be problematic. Determining the sex of the tropical semiaquatic Varanus mertensi, a species with significant tail musculature, is difficult using traditional hand pressure-induced hemipenile eversion. As an alternative, we propose an extended technique for identifying the sex of adult V. mertensi. We recommend using a combination of the traditional field-based hemipenile eversion technique and a measurement of the ratio of androgens to estradiol in plasma samples taken from field-captured animals. PMID- 15891074 TI - Structure of msj-1 gene: a comparative analysis. AB - Msj-1 gene encodes a DnaJ protein highly expressed in spermatids and spermatozoa of both rodents and amphibians. We isolated and characterized the msj-1 gene in mice. A bioinformatic approach was then used to predict the putative promoter region, chromosomal localization, and its presence in the human genome. The analysis of msj-1 genomic sequence revealed that msj-1 is an intronless gene. Interestingly, two regions (A and B, separated by 10,682 bp) on human chromosome 2 having respectively 78% and 77% nucleotide identity with the murine msj-1 coding region were identified. This suggests the existence of an msj-1-like gene also in humans. PMID- 15891075 TI - Characterization of an RFamide-related peptide orphan GPCR in C. elegans. AB - We cloned and characterized an orphan FMRFamide-related peptide (FaRP) GPCR in Caenorhabditis elegans. We synthesized numerous structurally different FaRPs that were found in the C. elegans genome by bioinformatic analysis and used them to screen cells expressing the C26F1.6 receptor. Two peptides ending in M(orL)VRFamide elicited a calcium response in receptor-expressing mammalian Chinese hamster ovary cells. The response was dose-dependent and appeared to be very specific; that is, none of the other FaRPs were active, not even closely related peptides also ending in M(orL)VRFamide, which are encoded by the same peptide precursor. Pharmacological profiling with a truncated series of the most active peptide revealed that the full peptide sequence is necessary for receptor activation. PMID- 15891076 TI - Incapacity of response to disulfide-reducing agent in Triton X-100-treated oocytes of starfish, Asterina pectinifera. AB - Resumption of meiosis in starfish oocytes is induced by the natural maturation inducing hormone, 1-methyladenine (1-MeAde). Oocyte maturation is also induced by the disulfide-reducing agent, dithiothreitol (DTT). Previous studies have shown that 1-MeAde controls meiosis by interacting with its receptors, which are located exclusively on oocyte plasma membrane. However, little is known about the mechanism of oocyte maturation induced by DTT. Thus, this study examined whether DTT interacts with 1-MeAde receptors to induce oocyte maturation. When oocytes were treated with Triton X-100, they failed to respond to 1-MeAde and DTT. Although the Triton X-100-treated oocytes recovered the capacity to respond to 1 MeAde during incubation in seawater, they remained unresponsive to DTT during seawater incubations. These results suggest that DTT does not interact with 1 MeAde receptors to induce oocyte maturation in starfish. It is possible that a protein essential for mediating DTT-induced maturation is eliminated from the oocytes surface following Triton X-100 treatment. PMID- 15891077 TI - Cloning and characterization of ecdysone receptor and ultraspiracle cDNAs from Spodoptera litura. AB - The insect molting hormone, ecdysone, induces a series of events critical for ecdysis, beginning with the formation of a complex between ecdysteroid receptor (EcR) and its partner protein, ultraspiracle (Usp). In the present study, we cloned cDNAs encoding EcR and Usp by RT-PCR and 5'-/3'-RACE from the common cutworm, Spodoptera litura, a destructive agricultural pest insect in East Asia. Hexahistidine-tagged recombinant EcR and Usp of S. litura were expressed in E. coli, and the proteins, after refolding, exhibited the specific binding to an ecdysone analogue in vitro. PMID- 15891078 TI - Ligand-dependent protein interactions of the estrogen receptors using the yeast two-hybrid system. AB - The protein-interacting ability of estrogen receptor beta (ERbeta) and estrogen receptor alpha (ERalpha) from Atlantic salmon (Salmo salar) was studied using the yeast two-hybrid system. The salmon ERbeta shared features with other nuclear receptors regarding dimerization, forming a homodimer and heterodimer with ERalpha. The activation of salmon ERs by estradiol-17beta was shown to be necessary for dimerization. Furthermore, the xenoestrogens, 4-nonylphenol and bisphenol A, were shown to promote ER dimerization in the yeast two-hybrid system. PMID- 15891079 TI - Genome duplication-driven evolution of gene families: insights from the formation of the insulin family. AB - The aim of this study was to outline the sequence of events that gave rise to the members of the insulin gene family in chordates. As part of our research, we looked for the chromosomal localization of insulin family members in the human genome. PMID- 15891080 TI - Involvement of adenylyl cyclase signaling mechanism in insulin and IGF-I coregulation of fundamental cell processes. AB - The discovery of the adenylyl cyclase signaling mechanism (ACSM) of insulin and related peptides allows for advancing the hypothesis of its participation in regulatory action on cell growth, apoptosis, and metabolism. The following evidences were obtained: (1) The order of efficiency of insulin and IGF-I growth promoting action and their antiapoptotic effect coincides with that of the peptide-activating action on AC. (2) The growth-promoting and antiapoptotic effects of insulin and IGF-I are distinctly reproduced by cAMP. (3) The anabolic effects of insulin and IGF-I activation on glycogen synthesis and the pentosophosphate pathway are inhibited by cAMP. PMID- 15891081 TI - The activities of antioxidant enzymes and monoamine oxidase and uncoupling protein 1 content in brown fat of hypo- and hyperthyroid rats. AB - We have studied the activities of antioxidant enzymes (AOE), namely, copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), and catalase (CAT), and the activity of catecholamine-degrading enzyme monoamine oxidase (MAO) and uncoupling protein 1 (UCP1) content in brown fat (BF) of hypo- and hyperthyroid rats. We found that hypothyroidism decreased BF UCP1 content and increased MAO, MnSOD, and CAT activities. T3 increased UCP1 content and MnSOD activity and decreased CuZnSOD, MAO, and CAT activities, while T4 significantly altered (decreased) only CAT activity. This study shows that UCP1 content and MAO and AOE activities in rat BF are notably affected by changed thyroid status. PMID- 15891082 TI - Sea bream (Sparus auratus) estrogen receptors: phylogeny and tissue distribution. AB - Most estrogen actions are mediated by two estrogen receptor subtypes (ERalpha and ERbeta). While a single ERbeta appears to be present in higher vertebrates, two forms of ERbeta encoded by different genes have recently been isolated in some teleost fish species. We investigated whether this also applies to the hermaphrodite sparid sea bream (Sparus auratus) and cloned a second ERbeta (sbERbeta2) in this species. We have also compared the tissue distribution of the three receptors as yet identified in sea bream, designated sbERalpha, sbERbeta1, and sbERbeta2. PMID- 15891083 TI - Fish community characterization in two stretches upstream and downstream of the Lambro River confluence with the Po River. AB - This study aimed to characterize the fish community in two stretches of the Po River, upstream and downstream of the Lambro River confluence, to verify the environmental effects of the presence of endocrine disrupters (EDs). More than 5000 specimens were captured, identified at the species level, and recorded for biometric parameters. Plasma, liver, and gonad fragments were sampled for histological and biochemical analyses. Few specimens belonging to carp and barbel showed gonads with the concomitant presence of male and female tissues and altered plasma levels of vitellogenin and sex steroids; thus, these species will be considered for successive environmental studies. PMID- 15891084 TI - Activity of purine catabolism enzymes during the reproductive cycle of male and female brown trout (Salmo trutta). AB - In order to detect predicted changes in purine catabolism during the annual reproductive cycle of brown trout, we measured the activity of xanthine dehydrogenase, urate oxidase, and allantoinase. In kidney, only xanthine dehydrogenase was detected. In female liver and kidney, the activity of these enzymes was higher in May and decreased during vitellogenesis, with urate oxidase being undetectable in this period. In male liver, a similar variation pattern was found; however, in kidney, high activities were found in both May and December. These results suggest an influence of sex hormones in trout purine catabolism, especially in females. PMID- 15891085 TI - Evolution of the thyroid hormone distributor protein transthyretin in microbes, C. elegans, and vertebrates. AB - Transthyretin (TTR) is an extracellular thyroid hormone distributor protein in vertebrates, whose structure has been highly conserved between fish and humans. However, the ligand preferentially bound by TTR has changed during evolution from 3',3,5-L-triiodothyronine (T3) to 3',5',3,5-l-tetraiodothyronine (T4). We identified genes in the genomes of >50 species of nonvertebrates, which could code for TTR-like proteins. Molecular modeling suggested most would have similar 3D structures and electrostatic surface potentials to vertebrate TTRs. We amplified TTR-like genes from a C. elegans cDNA library, demonstrating that it is transcribed. We synthesized recombinant TTR-like proteins from S. dublin and C. elegans. These proteins form tetramers similarly to vertebrate TTRs, but their ligands remain elusive. PMID- 15891086 TI - Ghrelin induces growth hormone (GH) secretion via nitric oxide (NO)/cGMP signaling. AB - Ghrelin, a recently discovered 28-aa peptide, stimulates GH release through a mechanism involving PLC- and cAMP-related signaling pathways. Recently, nitric oxide (NO) and its mediator, cGMP, have been shown to be required for the response of somatotropes to various regulators (GHRH, somatostatin, leptin). Here, we explore the possible role of the NO synthase (NOS)/NO/guanylate cyclase (GC)/cGMP signaling pathway in ghrelin-induced GH release from cultured pig somatotropes using blockers or activators of this route. PMID- 15891087 TI - Neurotoxicity of polychlorinated biphenyls (PCBs) by disturbance of thyroid hormone-regulated genes. AB - PCBs are known as neurotoxic compounds. Part of this neurotoxicity could be due to an alteration of the expression of TH-regulated genes in brain. To identify such genes, brain protein extracts of hypo- and hyperthyroid as well as PCB treated embryos were compared by fluorescent 2D-DIGE. In total, we observed 109 differentially expressed proteins, of which 17 differed with both PCB and hypo- or hyperthyroid treatment. It was found that the interaction of PCBs with the expression of TH-regulated genes is congener-specific and that both hyperthyroidism- and hypothyroidism-related effects occur. PMID- 15891088 TI - Effects of a teleost tetraploidization on neuropeptide Y receptor gene repertoire in ray-finned fishes. AB - The ancestral vertebrate repertoire for neuropeptide Y receptor genes of the Y1 subfamily probably included four subtypes: Y1, Y4, Y6, and Y8. There was probably a single gene in the Y5 category. Both Y1 and Y5 stimulate food intake in mammals. As the genome seems to have duplicated during the evolution of ray finned fishes, we have investigated the gene repertoire in species that diverged prior to the appearance of teleosts, as well as a basal teleost and a shark. Our results show that the genes Y1, Y5, and Y6, which are missing in many teleosts, are present in basal actinopterygians. These dramatic alterations of the teleost receptor repertoire may be related to the tetraploidization in a teleost ancestor. PMID- 15891089 TI - Unusual genomic structure: melanocortin receptors in Fugu. AB - The melanocortin (MC) receptors are found in five subtypes in mammals and chicken, while recent studies have shown that the Fugu (Takifugu rubripes) genome has only four MC receptors and the zebrafish genome has six subtypes. The MC3 receptor seems to be missing from the two closely related pufferfishes, Fugu and Tetraodon (Tetraodon nigroviridis). The MC2 and MC5 receptors in the pufferfish have introns. Moreover, these two receptors are found in a tandem that is remarkably conserved in several vertebrate species. Here, we speculate about the genomic origin of the MC receptors. PMID- 15891090 TI - Pleiotropic action of insulin-like peptides of mollusk, Anodonta cygnea. AB - Insulin-related peptides (IRPs) from ganglions of mollusk, Anodonta cygnea, were purified and characterized (IRP1-IRP13) using insulin and insulin-like growth factor-I (IGF-I) radioreceptor test systems. The IRPs were able to bind to insulin and IGF-I receptors. Dose-dependent curve slopes indicated that most IRPs bind with higher affinity to IGF-I receptors than to insulin receptors. The IRP regulatory action on the activity of the adenylyl cyclase signal system showed that these peptides stimulated adenylyl cyclase and GTP-binding activity of G proteins to the same extent as insulin and IGF-I. The data obtained suggest polyfunctional IRP action that apparently is determined by the molecular structure of individual isoforms. PMID- 15891091 TI - Sensitivity of adenylyl cyclase signaling system of the mollusk Anodonta cygnea ganglions to serotonin and adrenergic agonists. AB - For the first time, the adenylyl cyclase system (ACS) sensitive to biogenic amines in the mollusk Anodonta cygnea ganglions was revealed and characterized. Serotonin and isoproterenol stimulated AC activity and GTP-binding activity of heterotrimeric G-proteins. The AC-stimulating action of serotonin and isoproterenol was blocked by cyproheptadine and adrenergic antagonists, respectively. Using synthetic C-terminal peptides of G-protein alpha-subunit, it was shown that the biogenic amines realized their action on the ACS through different G-protein types: serotonin and isoproterenol activate G(s)-protein, while adrenaline preferably activates G(i)-protein. PMID- 15891092 TI - A survey on the expression of IGF-I in the early developing bony fish with special emphasis on the tilapia, Oreochromis niloticus. AB - The present study investigates the expression of IGF-I in the early developing tilapia (Oreochromis niloticus). IGF-I was detected very early in ontogeny (4-5 days postfertilization, DPF), first in liver and in organs involved in growth and metabolism, thus suggesting a high physiological impact of IGF-I in growth, metabolism, and organogenesis. PMID- 15891093 TI - Stimulation of midgut stem cell proliferation and differentiation by insect hormones and peptides. AB - Stem cells derived from midguts of the caterpillar, Spodoptera littoralis, can be induced to multiply and differentiate in vitro. Ecdysone (E) and 20 hydroxyecdysone (20E) had a concentration-dependent effect: E was more active in cell proliferation and 20E in differentiation. Ecdysteroid receptors in midgut stem cell nuclei were stained with the antibody 9B9. In addition, alpha arylphorin and four midgut differentiation factors (MDF) specifically stimulated proliferation and differentiation of stem cells, respectively. The activity of a panel of peptide growth factors and hormones on growth and metamorphosis of the insect midgut is discussed. PMID- 15891094 TI - Ray-fin fish tetraploidization gave rise to pufferfish duplicates of NPY and PYY, but zebrafish NPY duplicate was lost. AB - We have used sequence information and gene location to identify NPY family genes in the pufferfish, Takifugu rubripes (fugu), and zebrafish. Fugu has two copies of NPY, presumably resulting from the ray-fin fish tetraploidization. Zebrafish has probably lost one of the copies. Both species have two copies of PYY, the second of which was previously named PY. The two fugu NPY genes are predominantly expressed in brain. The two PYY genes are expressed in a broad range of tissues including brain and gonads. Thus, the NPY system appears to be more complex in teleosts than in tetrapods. PMID- 15891095 TI - Molecular evolution and subfunctionalization of Nkx2.1 in the rainbow trout. AB - Because specification of duplicated genes is considered to be a major driving force for diversity and evolution, it is important to understand the function of the duplicated genes. In mammals, Nkx2.1 is essential for the differentiation of thyroid follicular cells and for the transcription of thyroid-specific genes. We have cloned four distinct Nkx2.1 cDNAs from the rainbow trout, Oncorhynchus mykiss. In the present article, we will characterize the trout Nkx2.1 mRNAs and show their tissue distribution, their transcriptional activities, and their expression during early development. PMID- 15891096 TI - Immunocytochemical and phylogenetic distribution of aquaporins in the frog ventral skin and urinary bladder. AB - We recently cloned three cDNAs encoding frog aquaporin (AQP-h1, BAC07470; AQP-h2, BAC82379; and AQP-h3, BAC07471) from the ventral pelvic skin of the tree frog, Hyla japonica. The present study demonstrated that Hyla AQP-h2 was translocated from cytoplasmic pools to the apical plasma membranes of the granular cells in the bladder after antidiuretic hormone stimulation and that Hyla AQP-h2 and AQP h3 behaved similarly in the ventral pelvic skin. Further, we found that terrestrial and tree frogs, but not aquatic and semiterrestrial-adapted frogs, absorbed water from their ventral pelvic skin by AQP-h3-like protein in concert with AQP-h2-like protein. PMID- 15891097 TI - Linkage mapping of the [Pro2]somatostatin-14 gene in zebrafish: evolutionary perspectives. AB - Radiation hybrid mapping assigned the zebrafish [Pro(2)]somatostatin-14 (also termed somatostatin 2; SS2) gene to linkage group 23 of the zebrafish genome, close to the marker nadl1.2. Comparative genomic analysis revealed conserved syntenies of the SS2 gene locus with part of the human 1p36 region, where the cortistatin gene is located. This observation strongly suggests that the SS2 gene in nonmammalian species and the cortistatin gene in mammals are orthologous. PMID- 15891098 TI - Ontogeny of 3 beta-hydroxysteroid dehydrogenase and 5 alpha-reductase in the frog brain. AB - The distribution of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 5alpha reductase (5alpha-R) has been studied in the frog brain during development. Soon after hatching, 3beta-HSD- and 5alpha-R-immunoreactive (ir) cells appeared first in the olfactory bulb and in the rhombencephalon. Subsequently, 3beta-HSD-ir cells were seen in the hypothalamus and cerebellum, whereas 5alpha-R-ir cells were visualized in the pallium, preoptic nucleus, posterocentral nucleus, cerebellum, and pituitary gland. At stages XIII-XVIII, additional 3beta-HSD- and 5alpha-R-ir cells appeared in several regions of the telencephalon, diencephalon, and mesencephalon. At stages XIX-XXI, the number of 5alpha-R-ir cells increased in the preoptic nucleus. These observations indicate that biosynthesis of biologically active steroids occurs in the brain of tadpoles, suggesting that neurosteroids may play a role in brain development. PMID- 15891099 TI - Calcium influx through voltage-operated calcium channels is required for proopiomelanocortin protein expression in Xenopus melanotropes. AB - Melanotrope cells of Xenopus laevis generate transitory increases in intracellular Ca(2+), known as Ca(2+) oscillations. These oscillations arise from the influx of Ca(2+) through voltage-operated Ca(2+) channels (VOCCs). Such oscillations are the driving force for secretion of a-melanophore-stimulating hormone (alpha-MSH) from the cell. The influx of Ca(2+) through VOCCs initiates the mobilization of intracellular Ca(2+) to generate a Ca(2+) wave. The function of the Ca(2+) wave in the melanotrope is unknown, but its presence in the nucleus suggests a role in the regulation of gene expression, perhaps that of proopiomelanocortin (POMC), the precursor protein for alpha-MSH. To determine the possible function of Ca(2+) waves in Xenopus melanotropes, we addressed whether functional VOCCs, which are an established requirement for both secretion and Ca(2+) wave initiation, are also required to maintain POMC gene expression. PMID- 15891100 TI - ACE inhibitor captopril reduces ecdysteroids and oviposition in moths. AB - By using the selective ACE inhibitor captopril, we studied the effect of the angiotensin converting enzyme (ACE) on larval growth, metamorphosis, and reproduction in a lepidopteran species, the cotton leafworm, Spodoptera littoralis. Captopril was detrimental to adult formation and oviposition, and in female moths it elicited decreasing ecdysteroid levels, but increasing trypsin activities. Our results suggest that captopril downregulates oviposition by two independent pathways. Apparently, oviposition is influenced by a complex interaction of ACE, trypsin activity, and ecdysteroid levels. PMID- 15891101 TI - Regulation of thyroid hormone availability by iodothyronine deiodinases at the blood-brain barrier in birds. AB - It is accepted that type II iodothyronine deiodinase (D2) is predominantly found in brain, where it maintains homeostasis of thyroid hormone (TH) levels. The current study describes the production of a polyclonal D2 antiserum and its use in the comparison of D2 protein distribution with that of type I (D1) and type III (D3) deiodinase protein in the chicken choroid plexus (CP). Immunocytochemistry showed high D2 protein expression in the epithelial cells of the CP, whereas the D1 and D3 proteins were absent. Furthermore, dexamethasone treatment led to an upregulation of the D2 protein in these cells. PMID- 15891102 TI - Differential proteomics for studying Drosophila immunity. AB - Here, we report the identification of proteins associated with the immune response of Drosophila by analyzing the hemolymph profiles after infection. Two dimensional difference gel electrophoresis was used to study the secretome in the hemolymph of Drosophila larvae. Shortly after induction with lipopolysaccharides, we identified 10 proteins, which we designated "Drosophila instantly released immune proteins". Infection with Micrococcus luteus or Saccharomyces cerevisiae induced 20 and 19 differential protein spots, respectively. Next to known immune proteins, new candidates that require further investigation were identified. PMID- 15891103 TI - Melatonin plays a crucial role in the regulation of rhythmic clock gene expression in the mouse pars tuberalis. AB - Circadian rhythms in physiology and behavior are driven by a central clock residing within the hypothalamic suprachiasmatic nucleus (SCN). Molecularly, the biological clock is based on the transcriptional/translational feedback loop of clock genes (mPer, mCry, Clock, and Bmal1). Circadian expression of clock genes is not limited to the SCN, but is found in many peripheral tissues. Peripheral rhythms depend on neuroendocrine/neuronal output from the SCN. Melatonin, the hormone of darkness, represents an important neuroendocrine output of the circadian clock. The hypophyseal pars tuberalis (PT) is one of the main target regions for melatonin. The aim of the study was to test whether mPer, mCry, Clock, and Bmal1 are rhythmically expressed in the mouse PT and how the absence of melatonin receptors affects clock gene expression. We analyzed clock gene expression by in situ hybridization and compared wild-type (WT), melatonin 1 receptor knockout (MT1 ko), and melatonin 2 receptor knockout (MT2 ko) mice. mPer1, mCry1, Clock, and Bmal1, but not mPer2 and mCry2, were rhythmically expressed in the PT of WT and MT2 ko mice. In the PT of MT1 ko mice, expression of mPer1, mCry1, Clock, and Bmal1 was dramatically reduced. We conclude that melatonin, acting through the MT1 receptor, is an important regulator of rhythmic clock gene expression in the mouse PT. PMID- 15891104 TI - Brain-derived neurotrophic factor in the hypothalamo-hypophyseal system of Xenopus laevis. AB - Brain-derived neurotrophic factor (BDNF) is immunocytochemically demonstrated in the magnocellular nucleus and neural lobe of the pituitary gland of the amphibian Xenopus laevis. Immunoelectron microscopy shows BDNF in secretory granules of type A neurohemal axon terminals in the neural lobe of pituitary gland. It is proposed that BDNF released from the neural lobe acts as a neurohormone stimulating the secretory activity of the melanotrope cells in the intermediate pituitary lobe. PMID- 15891105 TI - Glomerular effects of AVT on the in situ perfused trunk preparation of the dogfish. AB - The effects of arginine vasotocin on the pattern of glomerular perfusion in an elasmobranch fish were examined using an in situ perfused renal trunk preparation. A significant antidiuresis was coupled with a marked reduction in the proportion of filtering glomeruli (86%-25%) and an increase in the proportion of perfused but not filtering (10%-53%) and non-perfused glomeruli (4%-22%). However, the reduction in filtering glomeruli was greater than predicted by measurements of urine flow rate and glomerular filtration rate alone, suggesting that alterations in single nephron glomerular filtration rate may occur. PMID- 15891106 TI - Brain expression of tyrosine hydroxylase and its regulation by steroid hormones in the European eel quantified by real-time PCR. AB - In the eel, dopamine inhibits pubertal development. To investigate the regulatory mechanisms involved, we developed a quantitative real-time RT-PCR assay for measurement of brain expression of tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of dopamine. TH expression was highest in the olfactory bulb, followed by the di-/mesencephalic areas and the telencephalon/preoptic area. TH expression in the optic lobes and hindbrain was low or below the detection limit. In vivo treatment with testosterone, but not estradiol, resulted in increased TH expression in the forebrain, except the optic tectum, but not in the hindbrain. The results were confirmed by in situ hybridization. PMID- 15891107 TI - Renal morphology of the euryhaline flounder (Platichthys flesus): distribution of arginine vasotocin receptor. AB - The current study characterized tubular segmentation of the European flounder nephron and localized the vasotocin receptor expression by immunohistochemistry. Flounder nephron was shown to comprise a prominent renal corpuscle, short neck segment, proximal tubule I, proximal tubule II, collecting tubule, and collecting duct. Using specific antibodies raised against flounder vasotocin receptor, specific V(1) receptor staining was detected within the glomeruli, the endothelial surface of the afferent and efferent arterioles, and the capillaries surrounding the collecting duct system. Immunostaining for the receptor was exclusively vascular and there did not appear to be a tubular component. PMID- 15891108 TI - Spontaneous propagating calcium waves underpin airway peristalsis in embryonic rat lung. AB - Prenatal airways from diverse species exhibit spontaneous peristaltic contractions (airway peristalsis). These contractile waves appear coupled to and may function to regulate prenatal lung growth. They are unaffected by atropine or tetrodotoxin but abolished by nifedipine. Nevertheless, the mechanisms by which these contractile waves are generated, regulated, and propagated remain obscure. Using calcium imaging and whole embryonic lung organ culture, we demonstrate for the first time that peristalsis of the embryonic airway is driven by spontaneous, regenerative, temperature-sensitive calcium (Ca2+) waves. These Ca2+ waves propagate between individual airway smooth muscle cells coupled via gap junctions, are likely to be action potential-mediated, and are dependent on not only extracellular calcium entry via L-type voltage-gated channels but also intracellular Ca2+ stores. Thus, if airway peristalsis regulates lung growth, these findings mean that airway smooth muscle Ca2+ waves in turn regulate prenatal lung morphogenesis. PMID- 15891109 TI - Proteinase-activated receptor-1 mediates elastase-induced apoptosis of human lung epithelial cells. AB - Apoptosis of distal lung epithelial cells plays a pivotal role in the pathogenesis of acute lung injury. In this context, proteinases, either circulating or leukocyte-derived, may contribute to epithelial apoptosis and lung injury. We hypothesized that apoptosis of lung epithelial cells induced by leukocyte elastase is mediated via the proteinase activated receptor (PAR)-1. Leukocyte elastase, thrombin, and PAR-1-activating peptide, but not the control peptide, induced apoptosis in human airway and alveolar epithelial cells as assessed by increases in cytoplasmic histone-associated DNA fragments and TUNEL staining. These effects were largely prevented by a specific PAR-1 antagonist and by short interfering RNA directed against PAR-1. To ascertain the mechanism of epithelial apoptosis, we determined that PAR-1AP, thrombin, and leukocyte elastase dissipated mitochondrial membrane potential, induced translocation of cytochrome c to the cytosol, enhanced cleavage of caspase-9 and caspase-3, and led to JNK activation and Akt inhibition. In concert, these observations provide strong evidence that leukocyte elastase mediates apoptosis of human lung epithelial cells through PAR-1-dependent modulation of the intrinsic apoptotic pathway via alterations in mitochondrial permeability and by modulation of JNK and Akt. PMID- 15891110 TI - Bone marrow-derived mesenchymal stem cells in repair of the injured lung. AB - We sought to determine whether an intact bone marrow is essential to lung repair following bleomycin-induced lung injury in mice, and the mechanisms of any protective effects conferred by bone marrow-derived mesenchymal stem cell (BMDMSC) transfer. We found that myelosupression increased susceptibility to bleomycin injury and that BMDMSC transfer was protective. Protection was associated with the differentiation of engrafted BMDMSC into specific and distinct lung cell phenotypes, with an increase in circulating levels of G-CSF and GM-CSF (known for their ability to promote the mobilization of endogenous stem cells) and with a decrease in inflammatory cytokines. In vitro, cells from injured, but not from normal, mouse lung produced soluble factors that caused BMDMSC to proliferate and migrate toward the injured lung. We conclude that bone marrow stem cells are important in the repair of bleomycin-injured lung and that transfer of mesenchymal stem cells protects against the injury. BMDMSC localize to the injured lung and assume lung cell phenotypes, but protection from injury and fibrosis also involves suppression of inflammation and triggering production of reparative growth factors. PMID- 15891111 TI - Using a microfluidic device for 1 microl DNA microarray hybridization in 500 s. AB - This work describes a novel and simple modification of the current microarray format. It reduces the sample/reagent volume to 1 microl and the hybridization time to 500 s. Both 20mer and 80mer oligonucleotide probes and singly labeled 20mer and 80mer targets, representative of the T-cell acute lymphocytic leukemia 1 (TAL1) gene, have been used to elucidate the performance of this hybridization approach. In this format, called shuttle hybridization, a conventional flat glass DNA microarray is integrated with a PMMA microfluidic chip to reduce the sample and reagent consumption to 1/100 of that associated with the conventional format. A serpentine microtrench is designed and fabricated on a PMMA chip using a widely available CO2 laser scriber. The trench spacing is compatible with the inter-spot distance in standard microarrays. The microtrench chip and microarray chip are easily aligned and assembled manually so that the microarray is integrated with a microfluidic channel. Discrete sample plugs are employed in the microchannel for hybridization. Flowing through the microchannel with alternating depths and widths scrambles continuous sample plug into discrete short plugs. These plugs are shuttled back and forth along the channel, sweeping over microarray probes while re-circulation mixing occurs inside the plugs. Integrating the microarrays into the microfluidic channel reduces the DNA-DNA hybridization time from 18 h to 500 s. Additionally, the enhancement of DNA hybridization reaction by the microfluidic device is investigated by determining the coefficient of variation (CV), the growth rate of the hybridization signal and the ability to discriminate single-base mismatch. Detection limit of 19 amol was obtained for shuttle hybridization. A 1 mul target was used to hybridize with an array that can hold 5000 probes. PMID- 15891112 TI - Proof of concept for microarray-based detection of DNA-binding oncogenes in cell extracts. AB - The function of DNA-binding proteins is controlled not just by their abundance, but mainly at the level of their activity in terms of their interactions with DNA and protein targets. Moreover, the affinity of such transcription factors to their target sequences is often controlled by co-factors and/or modifications that are not easily assessed from biological samples. Here, we describe a scalable method for monitoring protein-DNA interactions on a microarray surface. This approach was designed to determine the DNA-binding activity of proteins in crude cell extracts, complementing conventional expression profiling arrays. Enzymatic labeling of DNA enables direct normalization of the protein binding to the microarray, allowing the estimation of relative binding affinities. Using DNA sequences covering a range of affinities, we show that the new microarray-based method yields binding strength estimates similar to low-throughput gel mobility shift assays. The microarray is also of high sensitivity, as it allows the detection of a rare DNA-binding protein from breast cancer cells, the human tumor suppressor AP-2. This approach thus mediates precise and robust assessment of the activity of DNA-binding proteins and takes present DNA-binding assays to a high throughput level. PMID- 15891113 TI - Identification of brassinosteroid-related genes by means of transcript co response analyses. AB - The comprehensive systems-biology database (CSB.DB) was used to reveal brassinosteroid (BR)-related genes from expression profiles based on co-response analyses. Genes exhibiting simultaneous changes in transcript levels are candidates of common transcriptional regulation. Combining numerous different experiments in data matrices allows ruling out outliers and conditional changes of transcript levels. CSB.DB was queried for transcriptional co-responses with the BR-signalling components BRI1 and BAK1: 301 out of 9694 genes represented in the nasc0271 database showed co-responses with both genes. As expected, these genes comprised pathway-involved genes (e.g. 72 BR-induced genes), because the BRI1 and BAK1 proteins are required for BR-responses. But transcript co-response takes the analysis a step further compared with direct approaches because BR related non BR-responsive genes were identified. Insights into networks and the functional context of genes are provided, because factors determining expression patterns are reflected in correlations. Our findings demonstrate that transcript co-response analysis presents a valuable resource to uncover common regulatory patterns of genes. Different data matrices in CSB.DB allow examination of specific biological questions. All matrices are publicly available through CSB.DB. This work presents one possible roadmap to use the CSB.DB resources. PMID- 15891114 TI - Clustering and conservation patterns of human microRNAs. AB - MicroRNAs (miRNAs) are approximately 22 nt-long non-coding RNA molecules, believed to play important roles in gene regulation. We present a comprehensive analysis of the conservation and clustering patterns of known miRNAs in human. We show that human miRNA gene clustering is significantly higher than expected at random. A total of 37% of the known human miRNA genes analyzed in this study appear in clusters of two or more with pairwise chromosomal distances of at most 3000 nt. Comparison of the miRNA sequences with their homologs in four other organisms reveals a typical conservation pattern, persistent throughout the clusters. Furthermore, we show enrichment in the typical conservation patterns and other miRNA-like properties in the vicinity of known miRNA genes, compared with random genomic regions. This may imply that additional, yet unknown, miRNAs reside in these regions, consistent with the current recognition that there are overlooked miRNAs. Indeed, by comparing our predictions with cloning results and with identified miRNA genes in other mammals, we corroborate the predictions of 18 additional human miRNA genes in the vicinity of the previously known ones. Our study raises the proportion of clustered human miRNAs that are <3000 nt apart to 42%. This suggests that the clustering of miRNA genes is higher than currently acknowledged, alluding to its evolutionary and functional implications. PMID- 15891115 TI - The recent transfer of a homing endonuclease gene. AB - The myxomycete Didymium iridis (isolate Panama 2) contains a mobile group I intron named Dir.S956-1 after position 956 in the nuclear small subunit (SSU) rRNA gene. The intron is efficiently spread through homing by the intron-encoded homing endonuclease I-DirI. Homing endonuclease genes (HEGs) usually spread with their associated introns as a unit, but infrequently also spread independent of introns (or inteins). Clear examples of HEG mobility are however sparse. Here, we provide evidence for the transfer of a HEG into a group I intron named Dir.S956-2 that is inserted into the SSU rDNA of the Costa Rica 8 isolate of D.iridis. Similarities between intron sequences that flank the HEG and rDNA sequences that flank the intron (the homing endonuclease recognition sequence) suggest that the HEG invaded the intron during the recent evolution in a homing-like event. Dir.S956-2 is inserted into the same SSU site as Dir.S956-1. Remarkably, the two group I introns encode distantly related splicing ribozymes with phylogenetically related HEGs inserted on the opposite strands of different peripheral loop regions. The HEGs are both interrupted by small spliceosomal introns that must be removed during RNA maturation. PMID- 15891116 TI - Chromatin assembly factor Asf1p-dependent occupancy of the SAS histone acetyltransferase complex at the silent mating-type locus HMLalpha. AB - Transcriptional repression of the silent mating-type loci HMLalpha and HMRa in Saccharomyces cerevisiae is regulated by chromatin structure. Sas2p is a catalytic subunit of the SAS histone acetyltransferase (HAT) complex. Although many HATs seem to relieve chromosomal repression to facilitate transcriptional activation, sas mutant phenotypes include loss of SIR1-dependent silencing of HMLalpha. To gain insight into the mechanism of the SAS complex mediated silencing at HMLalpha, we investigated the expression and chromatin structure of the alpha2 gene in the HMLalpha locus. We found that deletion of SAS2 in combination with a null allele of SIR1 changed the chromatin structure of the precisely positioned nucleosome, which includes the mRNA start site of the alpha2 gene and derepressed alpha2 transcription. The Sas2p HAT domain was required for this silencing. Furthermore, chromatin immunoprecipitation analysis revealed that the SAS complex was associated with the HMLalpha locus, and ASF1 (which encodes chromatin assembly factor Asf1p), but not SIR1 and SIR2, was necessary for this localization. These data suggest that the HAT activity and ASF1-dependent localization of the SAS complex are required for SIR1-dependent HMLalpha silencing. PMID- 15891117 TI - RNR1, a 3'-5' exoribonuclease belonging to the RNR superfamily, catalyzes 3' maturation of chloroplast ribosomal RNAs in Arabidopsis thaliana. AB - Arabidopsis thaliana chloroplasts contain at least two 3' to 5' exoribonucleases, polynucleotide phosphorylase (PNPase) and an RNase R homolog (RNR1). PNPase has been implicated in both mRNA and 23S rRNA 3' processing. However, the observed maturation defects do not affect chloroplast translation, suggesting that the overall role of PNPase in maturation of chloroplast rRNA is not essential. Here, we show that this role can be largely ascribed to RNR1, for which homozygous mutants germinate only on sucrose-containing media, and have white cotyledons and pale green rosette leaves. Accumulation of chloroplast-encoded mRNAs and tRNAs is unaffected in such mutants, suggesting that RNR1 activity is either unnecessary or redundant for their processing and turnover. However, accumulation of several chloroplast rRNA species is severely affected. High-resolution RNA gel blot analysis, and mapping of 5' and 3' ends, revealed that RNR1 is involved in the maturation of 23S, 16S and 5S rRNAs. The 3' extensions of the accumulating 5S rRNA precursors can be efficiently removed in vitro by purified RNR1, consistent with this view. Our data suggest that decreased accumulation of mature chloroplast ribosomal RNAs leads to a reduction in the number of translating ribosomes, ultimately compromising chloroplast protein abundance and thus plant growth and development. PMID- 15891119 TI - Irritable bowel syndrome: the need to exclude Dientamoeba fragilis. PMID- 15891120 TI - High prevalence of Bartonella quintana endocarditis in Sfax, Tunisia. AB - Bartonella quintana is a fastidious microorganism associated with blood culture negative endocarditis. In this study, 40 sera with cross-reactivity between Chlamydia species from patients from Sfax, Tunisia, were serologically tested for Bartonella. Thirteen sera were positive for Bartonella with IgG titers >/=1:800. Western blot and cross-absorption confirmed the diagnosis of Bartonella quintana endocarditis in 12 cases and Bartonella henselae endocarditis in 1 case. These sera were also positive by LightCycler nested PCR amplification for the rnpb (7 of 13) and fur (11 of 13) genes. Eleven patients had a definite diagnosis of endocarditis, which represents 9.8% of all endocarditis. Because Bartonella endocarditis seems to be very common in Tunisia, we suggest that its serology be performed systematically whenever endocarditis is suspected. PMID- 15891121 TI - Cutaneous fascioliasis: a case report in Vietnam. AB - A 40 year-old woman living in Gialai, Kontum, Vietnam, developed a red solid mass in the epigastric region. From ultrasound investigation, liver abscess and myositis of the intercostal muscle was diagnosed. Two weeks after treatment with antibiotics, the mass disappeared, but a migratory track developed in the right upper quadrant of the abdomen. An aspiration of the vesicular end of the serpiginous track showed a light brown, living worm that was later identified as an immature Fasciola sp. This is the first case report of cutaneous fascioliasis in the form similar to creeping eruption. PMID- 15891122 TI - Short report: evaluation of a self-detection tool for tapeworm carriers for use in public health. AB - The current study was designed to evaluate a tool for the self-identification of tapeworm carriers. Clinical and animal health care practitioners and schoolteachers were trained regarding the life cycle, risk factors, and control measures related to infection with Taenia solium. More than 120 small glass bottles with a few tapeworm segments fixed in formaldehyde and an instructional guide were distributed among all clinical practitioners (physicians and nurses) working in health centers. The guide contained 10 key points on how to ask questions about tapeworm infections. Information on taeniosis and cysticercosis was also provided to the general population via different media. Seven tapeworm carriers were recorded in the official epidemiology surveillance system the year previous to the study, compared with the year after the study, when 41 tapeworm carriers (37 Taenia saginata; 4 Taenia solium) were recorded. Six times more tapeworm carriers were notified after the study. All four persons with Taenia solium were treated, thereby eliminating the parasite and subsequently preventing any new cases of human and swine cysticercosis that might have arisen from them. PMID- 15891123 TI - Case report: human brain abscess due to a tetra-acetabulate plerocercoid metacestode (Cyclophyllidea). AB - A 38-year-old man living near Phnom Penh (Cambodia) was admitted to a hospital in Paris in June 2001 for a single episode of a generalized grand mal seizure. This episode was preceded by a 9-month history of headaches. Magnetic resonance imaging (MRI) of the head revealed a rounded lesion immediately ahead of the left central sulcus. The resected lesion was about 20 mm in diameter. Histologic examination revealed an elongated but unsegmented metacestode at the center of the lesion. Polymerase chain reaction (PCR) analysis was inconclusive due to formalin-based histologic processing of the tissue. Morphologic analysis based on the histologic sections revealed that the metacestode was a tetra-acetabulate plerocercoid of the order Cyclophyllidea, with a distinct rostellum and pseudosegmentation of the dorsoventrally flattened hindbody. This is the first report of a tetra-acetabulate plerocercoid from a human host and the first report of any cyclophyllidean plerocercoid from the human brain. After 6 weeks, the patient was asymptomatic, neurologic examination was normal, and the brain MRI showed only surgical cavitation. The patient returned to Cambodia. PMID- 15891124 TI - A randomized, double-blind, double-dummy, controlled dose comparison of thalidomide for treatment of erythema nodosum leprosum. AB - In a randomized, double-blind, double-dummy controlled study, 22 men with erythema nodosum leprosum (ENL) received six capsules containing either 100 mg (group A, n = 12) or 300 mg (group B, n = 10) of thalidomide daily for one week. A six-week, four capsules per day taper followed, in which group A received 50 mg/day of thalidomide in weeks 2 and 3, then dummy capsules in weeks 4 through 7, while group B had gradual decrements every two weeks. Both regimens caused comparable improvement in 19 patients at day 7 (group A [12 of 12] versus group B [7 of 10]; P = 0.08), but slower tapering in group B showed less re-emergence of ENL through week 7 (P = 0.02, versus group A). Most patients developed new lesions soon after stopping treatment. Slower tapering from a higher initial thalidomide dose may improve clinical ENL responses, but high recurrence rates after discontinuation indicates further assessment is needed to identify better tapering regimens. PMID- 15891125 TI - Relationship between Schistosoma japonicum and nutritional status among children and young adults in Leyte, the Philippines. AB - The objectives of this study were 1) to provide more accurate estimates of the relationship between Schistosoma japonicum infection and both protein energy malnutrition (PEM) and anemia through better adjustment for potential confounders such as socioeconomic status (SES) and geo-helminth infections and 2) to assess the role of occult blood loss in mediating S. japonicum-associated anemia. We examined cross-sectionally 729 individuals (86.7% S. japonicum-infected and 13.3% S. japonicum-uninfected) aged 7-30 years in Leyte, The Philippines. The main outcome measures were height-for-age Z-score (HAZ), body-mass-index Z-score (BMIZ), triceps skinfold Z-score, hemoglobin, and fecal occult blood loss. Multivariate models were created to assess the relationship between S. japonicum infection and nutritional status after adjusting for age, gender, other helminths, and SES. After controlling for confounders, intensity of S. japonicum infection was inversely related to hemoglobin in all age groups (P < 0.0001) and HAZ among children /= 0.92 and test-retest reliabilities ranging from 0.61 to 0.89. A household SES questionnaire was administered. A logistic regression model was used to quantify the association between performance in different cognitive domains (learning, memory, verbal fluency, and the Philippine Non-Verbal Intelligence Test) and helminth infections. After adjusting for age, sex, nutritional status, hemoglobin, and SES, S. japonicum infection was associated with poor performance on tests of learning (odds ratio [OR] = 3.04, 95% confidence interval [CI] = 1.1 6.9), A. lumbricoides infection was associated with poor performance on tests of memory (OR = 2.2, 95% CI = 1.04-4.7), and T. trichiura infection was associated with poor performance on tests of verbal fluency (OR = 4.5, 95% CI = 1.04-30). Helminth infection was associated with lower performance in three of the four cognitive domains examined in this study. These relationships remained after rigorous control for other helminths and important confounding covariates. PMID- 15891128 TI - Strongyloides stercoralis recombinant NIE antigen shares epitope with recombinant Ves v 5 and Pol a 5 allergens of insects. AB - A new recombinant protein (NIE) for immunodiagnosis of human Strongyloides infection has 13% to 18% amino acid identity with antigen 5 insect venom allergen, but the C-terminal segment of NIE showed highest identity with Ves v 5 (yellow jacket) and Pol a 5 (paper wasp). A rabbit polyclonal anti-NIE antibody identified a single band of NIE antigen as well as bands of Pol a 5 and Ves v 5 antigens, and mouse anti-Pol a 5 and anti-Ves v 5 sera reacted with recombinant NIE antigen by Western blot. A cyanogen bromide-digested C-terminal fragment of NIE was reactive with mouse anti-Ves v 5 and Pol a 5 antibodies as well as with rabbit anti-NIE serum. Although IgE and IgG antibodies from pooled sera from Strongyloides-infected patients reacted with Pol a 5 and Ves v 5 recombinant antigens on immunoblots, neither antigen inhibited human IgG reaction with NIE antigen in a competitive enzyme-linked immunosorbent assay. PMID- 15891129 TI - Epidemic malaria in the highlands of Papua New Guinea. AB - As part of a larger study into the epidemiology of malaria in the highlands of Papua New Guinea, outbreak investigations were carried out at the end of the 2002 rainy season in 11 villages situated between 1,400 and 1,700 meters above sea level that had reported epidemics. Locations and timing of these epidemics corresponded largely to those reported in the pre-control era of the 1960s and 1970s. On average, 28.8% (range = 10.3-63.2%) of people in each of the 11 villages were found to be infected with malaria. Plasmodium falciparum accounted for 59% of all identified infections and P. vivax for 34%. The majority (53%) of infections were symptomatic. Although symptomatic infections were most common in children 2-9 years of age (36%), even in adults a prevalence of 20% was observed. A comparison with earlier non-epidemic data in three of the villages without easy access to health care showed markedly increased levels of morbidity, with 6-10 fold increases in parasite prevalence, a 3-fold increase in both measured and reported fevers, and a 12-fold increase in enlarged spleens. The average hemoglobin levels were reduced by 2.3-3.5 g/dL, with a concurrent increase in moderate to severe anemia (hemoglobin level < 7.5 g/dL) from 0.0-3.3% to 3.8 18.4%. These massive increases in morbidity have devastating impact on the affected communities and highlight that malaria epidemics are a serious and increasing public health problem in the highlands of Papua New Guinea. PMID- 15891130 TI - Clinical manifestations of severe malaria in the highlands of southwestern Uganda. AB - Epidemics of malaria have occurred in highland areas of East Africa since the 1980s, but the clinical spectrum of severe malaria in these areas has not been described. Over a 17-month period from 2001 to 2002, we assessed 117 consecutive patients admitted to Kabale Hospital in highland Uganda who met the World Health Organization 2000 criteria for severe malaria. Sixty-six persons (56.4%) were age 5 years or older, and 51 (43.6%) were under 5 years of age. Fever, vomiting, and cough were the most frequent symptoms. Hepatomegaly and splenomegaly were infrequent. Prostration was the most frequent manifestation of severe malaria in children under 5 years of age (45.1%) and persons 5 years or older (65.2%), followed by respiratory distress (29.4%) and severe anemia (19.6%) in children under 5 years, and respiratory distress (15.2%) and impaired consciousness (13.6%) in persons 5 years or older. Strictly defined cerebral malaria was uncommon (3.4%). In a multivariate regression model, children under 5 years were more likely than persons 5 years or older to present with severe anemia (OR 5.2, 95% confidence interval [CI] 1.2-21.9) and respiratory distress (OR 3.5, 95% CI 1.3-11.1) and less likely to present with prostration (OR 0.3, 95% CI 0.1-0.7) and impaired consciousness (OR 0.2, 95% CI 0.0-0.9). In highland Uganda, severe malaria often occurs in persons older than 5 years of age. "Typical" signs like splenomegaly are frequently absent, prostration is the major manifestation, and other manifestations vary in frequency according to age. PMID- 15891131 TI - Efficacy and tolerability of artesunate plus sulfadoxine-pyrimethamine and sulfadoxine-pyrimethamine alone for the treatment of uncomplicated Plasmodium falciparum malaria in Peru. AB - To assist the Peruvian Ministry of Health in modifying the malaria treatment policy for their north Pacific coastal region, we conducted an in vivo efficacy trial of sulfadoxine-pyrimethamine (SP) and SP plus artesunate (SP-AS) for the treatment for uncomplicated Plasmodium falciparum infections. A total of 197 patients were randomized to therapy with either SP (25 mg/kg of the sulfadoxine component in a single dose on day 0) or a combination of SP plus AS (4 mg/kg on days 0, 1, and 2) and were followed for 28 days for symptoms and recurrence of parasitemia. No statistically significant differences between the two groups were observed on enrollment with respect to age, sex, history of malaria, or geometric mean parasite density. A total of 185 subjects completed the 28-day follow-up. Of the 91 subjects treated with SP alone, two had recurrences of parasitemia on day 7 and one on day 21. Of the 94 subjects treated with SP-AS, one had a recurrence of parasitemia on day 21. Fever and asexual parasite density decreased significantly more rapidly and the proportion of patients with gametocytemia on days 3-28 was significantly lower in subjects treated with combination therapy than in those who received SP alone. No severe adverse drug reactions were observed; however, self-limited rash and pruritus were significantly more common and an exacerbation of nausea, vomiting, and abdominal pain were observed significantly more frequently among patients who had received SP-AS. These results have contributed to a National Malaria Control Program decision to change to SP-AS combination therapy as the first-line treatment for uncomplicated P. falciparum malaria in northern coastal Peru in November 2001, making Peru the first country in the Americas to recommend this combination therapy. PMID- 15891132 TI - Sulfadoxine-pyrimethamine plus chloroquine or amodiaquine for uncomplicated falciparum malaria: a randomized, multisite trial to guide national policy in Uganda. AB - The use of combinations of inexpensive drugs for the treatment of malaria in Africa has been proposed as an interim policy while awaiting the widespread availability of more effective regimens. We compared sulfadoxine-pyrimethamine plus chloroquine or amodiaquine in three districts in Uganda. Patients aged 6 months or greater with uncomplicated falciparum malaria were enrolled and randomized to therapy. Safety, tolerability, and efficacy outcomes, adjusted by genotyping, were assessed over 28 days. Of 1,105 patients enrolled, 1,057 (96%) completed follow-up. For children less than 5 years old, the risk of clinical treatment failure adjusted by genotyping at the three sites ranged from 34% to 67% with chloroquine plus sulfadoxine-pyrimethamine and from 13% to 35% with amodiaquine plus sulfadoxine-pyrimethamine (risk differences 21-32%, P < 0.0001 at all sites). Serious adverse events were uncommon with both regimens. The risk of treatment failure with chloroquine plus sulfadoxine-pyrimethamine, the current standard in Uganda, was unacceptably high. Amodiaquine plus sulfadoxine pyrimethamine was significantly more efficacious; however, existing levels of resistance raises concern about the useful therapeutic life-span of this regimen. PMID- 15891133 TI - pfmdr1 genotyping and in vivo mefloquine resistance on the Thai-Myanmar border. AB - Molecular markers have been proposed as a method of monitoring malaria drug resistance and could potentially be used to prolong the life span of antimalarial drugs. Single nucleotide polymorphisms (SNPs) in the Plasmodium falciparum gene pfmdr1 and increased gene copy number have been associated with in vitro drug resistance but have not been well studied in vivo. In a prospective cohort study of malaria patients receiving mefloquine treatment on the Thai-Myanmar border, there was no significant association between either pfmdr1 SNPs or in vitro drug sensitivity and mefloquine resistance in vivo. Increased pfmdr1 gene copy number was significantly associated with recrudescence (relative risk 2.30, 95% CI 1.27 4.15). pfmdr1 gene copy number may be a useful surveillance tool for mefloquine resistant falciparum malaria in Thailand. PMID- 15891134 TI - Immune complex levels in children with severe Plasmodium falciparum malaria. AB - Malaria infection leads to the formation of circulating immune complexes. However, it is unclear whether these complexes play a role in the pathogenesis of complicated Plasmodium falciparum malaria. This study aimed at determining if there are differences in the levels of immune complexes between children with severe malaria-associated anemia and cerebral malaria and between each of these two groups and their respective uncomplicated symptomatic malaria or healthy asymptomatic controls. Children with severe malaria-associated anemia and cerebral malaria had significantly higher immune complex levels than their respective controls, but there were no significant differences in the levels between the two severe malaria groups. In addition, there was an inverse relationship between the hemoglobin levels and immune complex levels in the severe anemia controls, suggesting that immune complexes may contribute to erythrocyte destruction in these children. These results suggest that immune complex levels alone cannot account for the differences in the distinct clinical presentation between severe malaria-associated anemia and cerebral malaria. PMID- 15891135 TI - Colorimetric assay for screening compounds against Leishmania amastigotes grown in macrophages. AB - An estimated 12 million persons throughout the world suffer from the protozoan disease leishmaniasis. Current treatments have liabilities including poor activity against some forms of leishmaniasis, toxicity, or the need for parenteral administration. Higher throughput methods to screen chemical compounds are needed to facilitate the search for new antileishmania drugs. In the mammalian host, Leishmania parasites exist as amastigotes that replicate within macrophages. Therefore, an in vitro screening assay using intramacrophage amastigotes most closely represents the natural infection. We have transfected strains of Leishmania major and Leishmania amazonensis with the beta-lactamase gene, which catalyzes a colorimetric reaction with the substrate nitrocephin. The growth of these beta-lactamase-expressing Leishmania within macrophages was quantified in 96-well plates using an optical density plate reader, thus simplifying the methodology for scoring inhibitor assays. This simple and relatively inexpensive colorimetric assay helps improve throughput for screening compounds for antileishmania activity. PMID- 15891136 TI - Species assignation of Leishmania from human and canine American tegumentary leishmaniasis cases by multilocus enzyme electrophoresis in North Argentina. AB - Sixteen Leishmania stocks, 15 isolated from patients with cutaneous (CL), mucocutaneous (MCL), or recurrent cutaneous leishmaniasis, plus one from a dog with CL in Salta and Corrientes Provinces, Argentina, were studied by multilocus enzyme electrophoresis. Thirteen of the stocks from humans were grouped in two zymodemes; nine termed as KMS 1, four as KMS 2, and assigned to Leishmania (Viannia) braziliensis. Two additional stocks from CL cases expressed a KMS 4 enzyme profile, corresponding to L. (V.) guyanensis. Although the parasites from the dog were also assigned to L. (V.) braziliensis, its zymodeme, KMS 3, was not expressed in any of the current human isolates. The characterization of Leishmania from a dog was done for the first time in Argentina. The importance of the intraspecific polymorphism in the induction of clinical forms and in the host reservoir concept is briefly discussed, based on the zymodeme data of isolates from humans and dogs. The presence of L. (V.) guyanensis was confirmed in the country. PMID- 15891137 TI - Short report: serological evidence of West Nile virus activity in El Salvador. AB - Epizootics of encephalitis in El Salvador killed 203 equines between November 2001 and April 2003. During an investigation of the outbreaks, 18 (25%) of 73 serum samples collected from stablemates of deceased animals in 2003 had antibodies to West Nile virus. Ten of these infections were confirmed by plaque reduction neutralization tests, suggesting West Nile virus has extended its range and spread to Central America. PMID- 15891138 TI - Differential infectivities of o'nyong-nyong and chikungunya virus isolates in Anopheles gambiae and Aedes aegypti mosquitoes. AB - O'nyong-nyong virus (ONNV) and chikungunya virus (CHIKV) are closely related alphaviruses that cause human disease in Africa and Asia. Like most alphaviruses, CHIKV is vectored by culicine mosquitoes. ONNV is considered unusual as it primarily infects anopheline mosquitoes; however, there are relatively few experimental data to support this. In this study, three strains of ONNV and one strain of CHIKV were evaluated in Anopheles gambiae and Aedes aegypti mosquitoes and in four cell lines. As predicted, CHIKV was not infectious to An. gambiae, and we observed strain-variability for ONNV with respect to the ability of the virus to infect An. gambiae and Ae. aegypti. The species specificity in vivo was reflected by in vitro experiments using culicine and anopheline-derived cell lines. PMID- 15891139 TI - Serologic evidence of exposure of wild mammals to flaviviruses in the central and eastern United States. AB - Serosurveys were conducted to obtain flavivirus and West Nile virus (WNV) seroprevalence data from mammals. Sera from 513 small- and medium-sized mammals collected during late summer and fall 2003 from Colorado, Louisiana, New York, Ohio, and Pennsylvania were screened for flavivirus-specific antibodies. Sera samples containing antibody to flaviviruses were screened for WNV-specific antibodies by epitope-blocking enzyme-linked immunosorbent assays and confirmed with plaque reduction neutralization tests. Prevalence of WNV antibodies among study sites ranged from 0% to 42.8% among the mammal communities sampled. High prevalence rates for WNV were noted among raccoons (100%, with a very small sample size, N = 2), Virginia opossums (50.0%), fox squirrels (49.1%), and eastern gray squirrels (48.3%). The high WNV antibody prevalence noted for tree squirrels, the peri-domestic tendencies of several of these species, and their ease of observation could make these species useful sentinels for monitoring WNV activity within urban communities. PMID- 15891140 TI - Effect of mosquito midgut trypsin activity on dengue-2 virus infection and dissemination in Aedes aegypti. AB - The effect of mosquito midgut trypsins in dengue serotype 2 flavivirus (DENV-2) infectivity to Aedes aegypti was studied. Addition of soybean trypsin inhibitor (STI) in a DENV-2 infectious blood meal resulted in a 91-97% decrease in midgut DENV-2 RNA copies (qRT-PCR analysis). STI treatment also resulted in slower DENV 2 replication in the midgut, less DENV-2 E protein expression, and decreased dissemination to the thorax and the head. A second uninfected blood meal, 7 days after the STI-treated infectious meal, significantly increased DENV-2 replication in the midgut and recovered oogenesis, suggesting that the lower viral infection caused by STI was in part due to a nutritional effect. Mosquitoes fed DENV-2 digested in vitro with bovine trypsin (before STI addition) exhibited a transient increase in midgut DENV-2 4 days postinfection. Blood digestion and possibly DENV 2 proteolytic processing, mediated by midgut trypsins, influence the rate of DENV 2 infection, replication, and dissemination in Ae. aegypti. PMID- 15891141 TI - Autochthonous epidemic typhus associated with Bartonella quintana bacteremia in a homeless person. AB - Trench fever, a louse-borne disease caused by Bartonella quintana, is reemerging in homeless persons. Epidemic typhus is another life-threatening louse-borne disease caused by Rickettsia prowazekii and known to occur in conditions of war, famine, refugee camps, cold weather, poverty, or lapses in public health. We report the first case of seroconversion to R. prowazekii in a homeless person of Marseilles, France. This was associated with B. quintana bacteremia. Although no outbreaks of typhus have been notified yet in the homeless population, this disease is likely to reemerge in such situation. PMID- 15891142 TI - Short report: detection of Orientia tsutsugamushi in clinical samples by quantitative real-time polymerase chain reaction. AB - Orientia tsutsugamushi infection causes scrub typhus, a common zoonosis of rural Asia. Orientia tsutsugamushi was recently detected by a real-time quantitative polymerase chain reaction (qPCR) assay in animal specimens. We evaluated the same qPCR assay in specimens obtained from patients with serologically proven scrub typhus infections. The 47-kDa qPCR assay was more sensitive than was mouse inoculation; it was reactive in whole blood specimens from all 10 isolate positive patients and in 7 of 17 isolate-negative individuals (P = 0.003, Fisher's two-tailed exact test). As few as 1,076 O. tsutsugamushi copies/microL were detected in whole blood. Four of 7 sera from isolate-proven scrub typhus infections were also reactive by qPCR. The assay was unreactive in all 12 individuals without scrub typhus infection. This is the first demonstration of a sensitive and specific real-time qPCR assay for human scrub typhus infection. PMID- 15891143 TI - Ultrasonographic examination of Haitian children with lymphatic filariasis: a longitudinal assessment in the context of antifilarial drug treatment. AB - To assess the clinical findings associated with detection of adult Wuchereria bancrofti worms on ultrasound, 186 schoolchildren in a filariasis-endemic area of Haiti underwent physical and ultrasonographic examinations. The filaria dance sign (FDS) of adult W. bancrofti was detected in the inguinal and crural lymphatics of 28 (15%) children. FDS detection was more common in older children (P = 0.003) and in those with a history of inguinal lymph node inflammation (P = 0.002) or crural lymphadenopathy on physical exam (P = 0.01). Twenty-five FDS positive children were reexamined after three annual cycles of mass treatment for lymphatic filariasis (LF). The total number of adult worm nests detected by ultrasound decreased from 29 to 4 (P /= 50%) verified with DSA. CONCLUSION: Regardless of slight underestimation of carotid stenosis with CT angiography compared with rotational angiography, diagnostic performance of CT angiography with interactive interpretation proved to be good. Also, the method is highly sensitive for detection of carotid artery stenosis, indicating the suitability of CT angiography as a screening method for symptomatic patients. For hemodynamically significant stenoses revealed by CT angiographic screening, conventional angiography still seems to be necessary. PMID- 15891156 TI - Dynamic spin labeling angiography in extracranial carotid artery stenosis. AB - BACKGROUND AND PURPOSE: Similar to digital subtraction angiography, dynamic spin labeling angiography (DSLA) provides time-resolved measurements of the influx of blood into the cerebral vascular tree. We determined whether DSLA may help in assessing the degree of stenosis and whether it provides information about intracerebral collateralization and allows us to monitor the hemodynamic effects of vascular interventions. METHODS: We developed a segmented DSLA sequence that allowed the formation of images representing inflow delays in 41-ms increments. Thirty patients with unilateral carotid artery stenosis and 10 control subjects underwent DSLA. Arrival times of the labeled arterial blood bolus were measured in the carotid siphon (CS) and the middle cerebral artery (MCA) on both sides, and the corresponding side-to-side arrival time differences (ATDs) were calculated. ATDs before and after carotid endarterectomy or percutaneous angioplasty were studied in 10 patients. RESULTS: The degree of stenosis was significantly correlated with ATD in the cerebral vessels. Receiver operating characteristic analysis yielded a cutoff CS ATD of 110 ms to separate stenoses <70% from those > or =70%, with a sensitivity of 90% and a specificity of 67%. In one third of patients, ATD was higher in the MCA than in the CS; this finding suggested an absence of collateralization. Most patients had reduced ATD in the MCA. The degree of ATD reduction was regarded as a quantitative measure of collateralization. Successful intervention resulted in normalized ATDs. CONCLUSION: DSLA is a promising method that allowed us to noninvasively quantify the hemodynamic effect of extracranial carotid stenosis and the resulting intracranial collateralization. PMID- 15891157 TI - Carotid plaque composition and cerebral infarction: MR imaging study. AB - BACKGROUND AND PURPOSE: Besides the severity of carotid artery stenosis, atherosclerotic plaque composition is an important determinant of cerebral symptoms. We analyzed the relationship between the composition of the atherosclerotic plaque at the carotid artery bifurcation and ipsilateral ischemic cerebral lesions on MR imaging. METHODS: Forty-one patients with symptomatic carotid artery stenosis (>70%) underwent black-blood, fast spin-echo imaging of the carotid artery and turbo fluid-attenuated inversion recovery (t-FLAIR) imaging of the brain. Plaque regions with a relative decrease in signal intensity in the plaque from proton density-weighted (TE = 14 ms) to T2-weighted (TE = 50 ms) imaging were considered to be lipid cores. We assessed the number and location of infarcts in the ipsilateral cortex, basal ganglia, and centrum semiovale, and hyperintense white matter lesions on t-FLAIR images. RESULTS: Lipid in the atherosclerotic plaque at the carotid bifurcation was seen in 25 patients. Ipsilateral infarctions were seen in 22 (54%); most often, it involved the centrum semiovale. Patients with a lipid core had an ipsilateral infarct more often than patients without a lipid core (68% vs. 31%; P = .03). Centrum semiovale infarcts were more frequent (56% vs. 25%, P = .06) and the median number of centrum semiovale infarcts was higher P = .04) in patients with a lipid core than in patients without a lipid core. CONCLUSION: Ischemic cerebral lesions were common in patients with symptomatic carotid artery disease. Plaque composition, as assessed with MR imaging, is related to the presence and extent of ischemic cerebral lesions. PMID- 15891158 TI - Prediction of hemorrhagic transformation in acute ischemic stroke: role of diffusion-weighted imaging and early parenchymal enhancement. AB - BACKGROUND AND PURPOSE: MR imaging may help in predicting hemorrhagic transformation (HT) in acute ischemic stroke. Our purpose was to determine whether the lesion volumes on diffusion-weighted (DW) imaging, apparent diffusion coefficient (ADC) values, and early parenchymal enhancement are predictive of HT and to investigate the mechanism of the enhancement. METHODS: We retrospectively examined 55 patients with acute ischemic stroke who underwent gadolinium-enhanced MR imaging within 6 hours of symptom onset and follow-up CT or MR imaging within 72 hours. Intravenous thrombolysis was performed in 15 patients. DW imaging lesion volumes and ADC values were compared between patients with and those without HT. ADCs and perfusion parameters were compared between lesions with and those without parenchymal enhancement. RESULTS: Nineteen (34.5%) patients had HT (14 with hemorrhagic infarction, five with parenchymal hematoma). Patients with HT had decreased mean ADCs and large lesion volumes on DW imaging, but differences were not significant (P > .05). HT occurred in five patients (100%) with parenchymal enhancement, which corresponded to the site of HT. In enhancing lesions, the ADC ratio (0.76 +/- 0.06) was slightly higher and the delay in time to peak (0.10 +/- 2.79) was less than respective values in the rest of the ischemic lesion (0.66 +/- 0.06 and 8.79 +/- 4.86, respectively; P = .068). CONCLUSION: Early parenchymal enhancement is highly specific for HT and may be associated with early reperfusion and damage to the blood-brain barrier in ischemic tissue. DW imaging lesion volumes and ADC values had no strong relationship with HT. PMID- 15891159 TI - Vascular occlusion sites determine differences in lesion growth from early apparent diffusion coefficient lesion to final infarct. AB - BACKGROUND AND PURPOSE: Occlusion of major cerebral arteries is the primary source of tissue damage in ischemic stroke and the target of thrombolytic therapy. We hypothesized that large infarcts in more proximal vascular occlusions correspond with substantially increased ischemic lesions shown on initial apparent diffusion coefficient (ADC) maps. METHODS: Initial ADC lesions in 120 patients with acute ischemic stroke were analyzed within 6 hours of stroke onset. Patients were categorized on the basis of vascular occlusion, as shown on MR angiography. Lesion volumes were determined by using manual delineation (ADC(man)) and a threshold method for ADC values (<550 x 10(-9) mm(2)/s(-1), ADC(<550)). Infarct volumes were analyzed by using T2-weighted (n = 109) or CT (n = 11) images obtained on days 5-8. RESULTS: Median lesion volumes for ADC(<550), ADC(man), and infarcts, respectively, were as follows: proximal internal carotid artery (ICA)/middle cerebral artery (MCA) occlusions, 10, 23, and 32 cm(3); carotid-T occlusions, 11, 37, and 138 cm(3); MCA trunk occlusions, 11, 27, and 44 cm(3)); and MCA branch occlusions 8, 27, and 21 cm(3). Initial ADC lesion volumes were different only between the carotid T and the MCA branch (P < .05). On days 5 8, infarct volumes decreased from proximal to distal sites (P < .05), with the exception of MCA trunk versus proximal ICA/MCA occlusions. Recanalization rate in carotid-T occlusion was significantly lower than those of all other occlusion types. CONCLUSION: Initial ADC lesions can be small, even in patients with proximal vascular occlusions. These patients develop considerably large infarctions, suggesting a high potential for infarct growth. This growth might be averted with improved early recanalization of proximal vascular occlusions. PMID- 15891160 TI - Wallerian degeneration of the pontocerebellar fibers. AB - Two cases of pontine infarct with Wallerian degeneration (WD) of the pontocerebellar fibers are described. WD of pontocerebellar fibers, seen bilaterally along the transverse pontine fibers, is more visible in the middle cerebellar peduncles and extends into the white matter of the cerebellar hemispheres. Understanding the anatomy of the white matter and the temporal evolution of this degeneration is essential in identifying WD. PMID- 15891161 TI - Proton MR spectroscopy in Wilson disease: analysis of 36 cases. AB - BACKGROUND AND PURPOSE: Wilson disease (WD) is rare but one of the few metabolic disorders that can possibly benefit from effective available treatments. The literature regarding proton MR spectroscopy (MRS) in WD is scarce and controversial. The purpose of this study was to determine the brain metabolic changes due to WD by using MRS. To our knowledge, this is the first time that MRS was performed in such a large sample of patients with WD. METHODS: Thirty-six patients with WD and 37 healthy volunteers were examined with MRS in the parieto occipital cortex, frontal white matter, and basal ganglia (BG). Ratios of the following metabolites were calculated in relation to creatine (Cr): N acetylaspartate (NAA), choline (Cho), myo-inositol (mI), and glutamine/glutamate (Glx). The mean peak line width was measured on each spectrum. RESULTS: Compared with control subjects, patients with WD had significantly decreased NAA/Cr ratios in the three studied areas (P < .005) and an increased mI/Cr ratio in the BG (P < .001). Cho/Cr and Glx/Cr did not differ between the groups. The mean peak line in the BG was wider in patients than in control subjects. CONCLUSION: WD is unequivocally associated with MRS changes that could possibly be assigned to neuronal loss (in the three studied areas), to gliosis, and to iron and/or copper deposition in the BG. PMID- 15891162 TI - Metabolite phantom correction of the nonuniform volume-selection profiles in MR spectroscopic imaging: application to temporal lobe epilepsy. AB - BACKGROUND AND PURPOSE: In MR spectroscopic imaging (MRSI), the volume-selection profiles of metabolites differ from each other. These differences cause variations in metabolite intensities, which are particularly prominent when the hippocampi are evaluated. We hypothesize that the errors arising from these effects cause notable artifact when temporal lobe epilepsy (TLE) is lateralized with MRSI. METHODS: We examined a metabolite phantom, control subjects, and patients with TLE by using MRSI. We calculated the error arising from the different volume-selection profiles of metabolites in vitro and evaluated this correction in the examination of the control subjects and in the lateralization of epilepsy in the patients. RESULTS: Without a correction, a considerable error in the metabolite content existed, even deep inside the spectroscopic volume of interest. The result was false asymmetry (P < .008) in the hippocampi of control subjects. Among the 11 patients, TLE was correctly lateralized in three only after the correction was made, and in one, TLE was incorrectly lateralized. CONCLUSION: The volume-selection profiles of N-acetylaspartate, choline, and creatine differ enough to cause a significant error, even in the metabolite ratios, when patients with TLE are examined with MRSI. We propose a simple phantom method to correct for this error without a need to modify the pulse sequence. PMID- 15891163 TI - Rapid and fully automated visualization of subdural electrodes in the presurgical evaluation of epilepsy patients. AB - For rapid visualization of subdural electrodes with respect to cortical and subcortical structures, we describe a novel and fully automated method based on coregistration, normalization, optional cerebellum masking, and volume rendering of 3D MR imaging data taken before and after implantation. The key step employs the skull-stripped preimplantation image as a mask to also remove the skull in the postimplantation image. The extracted brain is presented in 3D with the electrodes directly visible by their susceptibility artifacts. Compared with alternative methods, ours is based on freely available software and does not require manual intervention. PMID- 15891164 TI - Single-dose contrast agent for intraoperative MR imaging of intrinsic brain tumors by using ferumoxtran-10. AB - BACKGROUND AND PURPOSE: Intraoperative MR imaging (IMRI) has advantages over conventional framed and frameless techniques. IMRI, however, also has some drawbacks, especially related to interpretation of gadolinium-enhanced intraoperative imaging resulting from surgically induced blood brain barrier injury, vascular changes, and hemorrhage. Ultra-small superparamagnetic iron particles like ferumoxtran-10 have a long plasma half-life and are trapped by reactive cells within the tumor. These trapped particles provide a method to demonstrate enhancing lesions without the artifact of repeat gadolinium administration in the face of blood brain barrier and vascular injury. METHODS: We present a review of the literature and the cases of two patients who underwent surgery in which IMRI with ferumoxtran-10 was used. RESULTS: Ultra-small superparamagnetic iron particles represent a method to demonstrate enhancing intrinsic brain tumors without the drawbacks of intraoperative gadolinium enhancement. These lesions appear even on low-field strength IMRI. Ferumoxtran 10, administered preoperatively, provides a stable imaging marker, even after surgical manipulation of the brain. CONCLUSION: Fermumoxtran-10 provides a way to lessen artifactual enhancement during IMRI related to the administration of gadolinium. PMID- 15891165 TI - Interhemispheric asymmetry of brain diffusivity in normal individuals: a diffusion-weighted MR imaging study. AB - BACKGROUND AND PURPOSE: Previous neuroimaging studies have suggested asymmetries in brain diffusivity may exist. The purpose of this study was to assess whether water diffusivity in deep gray matter structures shown by diffusion-weighted (DW) imaging differs between the right and left cerebral hemispheres in normal individuals. METHODS: Brain MR imaging was obtained in 23 healthy volunteers. A multisection image without diffusion weighting, and images with weighting applied in the read, phase, and section directions with a b-factor of 1000 s/mm(2) were collected. Diffusivity was computed separately in each direction, and the results were averaged to form mean diffusivity maps. Quantitative diffusivity values were obtained from the globus pallidus, putamen, caudate, thalamus, white matter, and CSF by using a standardized region of interest template. Interhemispheric differences were assessed by using a paired sample t test. RESULTS: Mean diffusivity was higher in the: left (mean +/- SD: 0.689 x 10(-3)+/- 0.069 x 10( 3)mm(2)/s) versus right (0.642 x 10(-3)+/- 0.071 x 10(-3)mm(2)/s) caudate (% difference, P value: 7.0%, P = .001); right (0.745 x 10(-3)+/- 0.053 x 10( 3)mm(2)/s) versus left (0.706 x 10(-3)+/- 0.050 x 10(-3)mm(2)/s) globus pallidus (5.2%, P < .001); left (0.720 x 10(-3)+/- 0.059 x 10(-3)mm(2)/s) versus right (0.674 x 10(-3)+/- 0.052 x 10(-3)mm(2)/s) putamen (6.4%, P < .001); right (0.750 x 10(-3)+/- 0.040 x 10(-3)mm(2)/s) versus left (0.716 x 10(-3)+/- 0.031 x 10( 3)mm(2)/s) thalamus (4.5%, P < .001). No significant right versus left difference was seen in the CSF (P = .291), anterior frontal white matter (P = .834), or centrum semiovale (P = .320). CONCLUSION: Gray matter diffusivity may differ between hemispheres of the brain in healthy individuals. Analysis of deep gray matter lesions requires caution, as statistically significant interhemispheric differences may not always be indicative of disease. PMID- 15891166 TI - Relationship between brain tissue oxygen tension and CT perfusion: feasibility and initial results. AB - BACKGROUND AND PURPOSE: Monitoring of intraparenchymal brain tissue oxygen tension (P(br)O(2)) is an emerging tool in neurocritical care. The purpose of this study was to determine if there is a relationship between CT perfusion (CTP) imaging parameters and P(br)O(2). METHODS: Nineteen patients underwent continuous P(br)O(2) monitoring with probes placed to target white matter in the cerebral hemisphere. Twenty-two CTP studies were performed at the level of the oxygen electrode, as identified on concurrent nonenhanced CT. CTP analysis software was used to measure mean transit time (MTT) and cerebral blood volume (CBV) and to derive cerebral blood flow (CBF) for a region of interest (ROI) surrounding the oxygen probe. For correlation, P(br)O(2) levels and other physiologic parameters were recorded at the time of CTP. RESULTS: P(br)O(2) values at the time of CTP were 2.7-54.4 mm Hg, MTT was 1.86-5.79 seconds, CBV was 1.18-8.76 mL/100 g, and CBF was 15.2-149.2 mL/100 g/min. MTT but not CBV or CBF was correlated with P(br)O(2) (r = -0.50, P = .017). MTT, CBV, or CBF were not correlated with other physiologic parameters, including mean arterial pressure, cerebral perfusion pressure, intracranial pressure, and fraction of inspired oxygen. On multivariable analysis, only P(br)O(2) was independently associated with MTT. CONCLUSION: CTP assessment of ROI surrounding an oxygen probe in the intraparenchymal brain tissue is feasible and showed a significant correlation between P(br)O(2) and MTT. Further studies are warranted to determine the role of CTP in assessing acute brain injury and whether it can be used to prospectively identify brain regions at risk for tissue hypoxia that should be targeted for advanced neuromonitoring. PMID- 15891167 TI - Regional cerebral blood flow after acetazolamide challenge in patients with dural arteriovenous fistula: simple way to evaluate intracranial venous hypertension. AB - BACKGROUND AND PURPOSE: Because venous hypertension determines the clinical severity of dural arteriovenous fistulas (DAVFs), evaluation of intracranial venous pressure is important in making decisions concerning treatment. We determined whether intracranial venous hypertension could be quantified by using an acetazolamide test in a manner that indicates whether treatment of the DAVF is necessary. METHODS: We enrolled 21 untreated patients: 11 with a sigmoid and/or transverse DAVF, six with a cavernous DAVF, two with an anterior cranial fossa DAVF, and two with a superior sagittal sinus DAVF. Cerebral hemodynamics were studied with stable-xenon CT. Patients were angiographically classified into three groups, and regional cerebral blood flow (rCBF) after acetazolamide challenge was compared between angiographic groups, between patients with and those without symptoms attributable to intracranial venous hypertension, and between preembolization and postembolization examinations. RESULTS: A high angiographic grade was associated with decreased resting rCBF and a blunted response to acetazolamide. Response to acetazolamide was more sensitive to venous hypertension, as angiographically assessed, than decreased resting rCBF. Resting rCBF and the increase in rCBF associated with acetazolamide were significantly lower in the symptomatic group than in the asymptomatic group. Treatment of the DAVF significantly enhanced the increased rCBF due to the acetazolamide challenge. CONCLUSION: Cerebral venous hypertension in DAVF reduced the response to acetazolamide, as shown on stable-Xe CT. Therefore, a patient with DAVF and a reduced rCBF response to acetazolamide requires treatment irrespective of his or her symptoms. PMID- 15891168 TI - High fractional anisotropy in brain abscesses versus other cystic intracranial lesions. AB - BACKGROUND AND PURPOSE: It is known that intracranial mass lesions are relatively isotropic on diffusion-weighted imaging. The purpose of this study is to report an unusually high fractional anisotropy (FA) and mean diffusivity (D(av)) in the cavity of the brain abscess compared with other cystic lesions. METHODS: We performed diffusion tensor imaging (DTI) in 12 patients with cystic intracranial lesions (pyogenic abscess, n = 5; cysticercus cysts, n = 2; and low-grade astrocytoma, n = 5). Mean FA, D(av) from the lesion core, perifocal edema, and corresponding contralateral normal-appearing regions were measured and compared for relative changes in these parameters. In the abscess cases, we placed regions of interest on areas with FA >0.2 and FA <0.2 to get FA and D(av) values. RESULTS: There were two patterns of FA values in the abscess cavity in all five patients. Part of the abscess showed mean FA = 0.440 +/- 0.135, with D(av) = (0.993 +/- 0.185) x 10(-3) mm(2)/s, whereas other parts had FA = 0.131 +/- 0.039 with D(av) = (0.824 +/- 0.183) x 10(-3) mm(2)/s. The cystic tumors and neurocysticercosis showed very high D(av) = (2.806 +/- 0.25, 2.654 +/- 0.35)x 10( 3) mm(2)/s, with low FA = (0.108 +/- 0.037, 0.08 +/- 0.01), respectively. CONCLUSION: Brain abscess cavity shows regions of increased FA values with restricted mean diffusivity compared with other cystic intracranial lesions. This information may prevent misinterpretation of the DTI information as white matter fiber bundle abnormalities associated with mass lesions. PMID- 15891169 TI - Diffusion-weighted imaging of fungal cerebral infection. AB - BACKGROUND AND PURPOSE: Diffusion-weighted imaging (DWI) is useful in diagnosing bacterial brain abscesses, but DWI features of fungal brain abscesses have not been characterized. Because fungal abscesses are not purulent, we hypothesized that their DWI characteristics are distinct from those of bacterial abscesses. METHODS: We reviewed clinical, neuropathologic and neuroimaging findings of patients with fungal brain infections due to Aspergillus (n = 6), Rhizopus (n = 1), or Scedosporium (n = 1) species. DWI and apparent diffusion coefficient (ADC) maps were obtained before definitive diagnosis and antifungal therapy. ADC ratios (lesion/contralateral white matter) were calculated. RESULTS: Two patients had a rapidly progressive, fatal course, with cerebritis and acute inflammation; fungal organisms were largely restricted to vessels. Lesions were predominantly nonenhancing and had heterogeneous foci of restricted diffusion. Six patients with subacute neurologic presentations had acute or chronic inflammation, capsule formation, focal necrosis, and fungal organisms disseminated throughout the lesion. Their abscesses were ring enhancing. In five, lesions had restricted diffusion in the central nonenhancing portions. The sixth patient had a lesion with a peripheral rim of restricted diffusion but elevated central diffusion; histopathology showed early abscess formation. Mean ADC for all lesions was 0.33 +/- 0.06 x 10(-3) mm(2)/s, with an average ADC ratio of 0.43. CONCLUSION: Fungal cerebral abscesses may have central restricted diffusion similar to that of bacterial abscesses but with histologic features of acute or chronic inflammation and necrosis rather than suppuration. Altered water diffusion in these lesions likely reflects highly proteinaceous fluid and cellular infiltration. PMID- 15891170 TI - Different signal intensities between intra- and extracranial components in jugular foramen meningioma: an enigma. AB - BACKGROUND AND PURPOSE: The purpose of this study was to evaluate retrospectively differences in MR signal intensity and contrast enhancement between intra- and extracranial components of jugular foramen meningioma (JFM). METHODS: MR studies of eight patients who underwent surgery for histologically confirmed JFM were reviewed retrospectively. Signal intensity differences between intra- and extracranial components of all eight JFMs on axial T1-, T2-, and postcontrast T1 weighted images were evaluated visually. In six of the eight JFMs, quantitative signal intensity evaluations were also performed by using relative signal intensity ratios of the intra- and extracranial components of JFM to CNS tissue at the same level. Paired t tests were used to evaluate differences in relative signal intensity ratios in each JFM between intra- and extracranial components. RESULTS: Both visual and quantitative signal intensity evaluations revealed that signal intensities of the intracranial component of JFM were significantly higher than those of the extracranial component on T1-, T2-, and postcontrast T1 weighted images. Results of relative signal intensity ratios were 0.89 +/- 0.04 versus 0.77 +/- 0.02 on T1-weighted images (P = .002); 1.66 +/- 0.28 versus 0.88 +/- 0.14 on T2-weighted images (P = .003); and 2.16 +/- 0.29 versus 1.77 +/- 0.26 on postcontrast T1-weighted images (P = .01). CONCLUSION: Intra- and extracranial components of JFM display different signal intensity and enhancement patterns. These differences may be related to histologic composition, and in particular, collagen content. PMID- 15891171 TI - Trigeminal ganglion and its divisions: detailed anatomic MR imaging with contrast enhanced 3D constructive interference in the steady state sequences. AB - BACKGROUND AND PURPOSE: Visualization of the trigeminal system is important for imaging diagnosis but technically challenging. We assessed how well the trigeminal ganglion, its rootlets, and its branches (V1, V2, and V3) are depicted on three high-resolution pulse sequences. METHODS: Twenty-two patients (44 sides) underwent nonenhanced 3D constructive interference in the steady state (CISS) MR imaging. Two of these patients and another 20 (44 sides) also underwent contrast enhanced 3D CISS and contrast-enhanced 3D time-of-flight (TOF) MR angiographic (MRA) imaging. Appearances of the ganglion, sinus ganglii, ganglion lip, and sensory and motor rootlets in the Meckel cave were assessed. RESULTS: The trigeminal ganglion was shown on enhanced 3D CISS images in all cases, on nonenhanced 3D CISS images in 77.3%, and on enhanced 3D TOF MRA images in 92.9%. Sinus ganglia and lips were demonstrated on 98% of enhanced 3D CISS images. Sensory rootlets were depicted with all 3D CISS sequences but no 3D TOF sequences. V1, V2, and V3 were displayed with all enhanced 3D TOF MRA sequences, 79.5-100% of enhanced 3D CISS sequences, and 0-50% of nonenhanced 3D CISS sequences. CONCLUSION: The enhanced 3D CISS sequence was best for displaying the trigeminal ganglion, sinus ganglii, and sinus lips, whereas the enhanced 3D TOF sequence best displayed the emerging V1, V2, and V3 roots. The enhanced 3D CISS sequence was most useful. Complete MR imaging evaluation of the trigeminal ganglion and roots is best performed by using enhanced 3D CISS and enhanced 3D TOF MRA sequences. PMID- 15891172 TI - A widespread pattern of cortical activations in patients at presentation with clinically isolated symptoms is associated with evolution to definite multiple sclerosis. AB - BACKGROUND AND PURPOSE: Movement-associated cortical changes have been detected at the earlier clinical stage multiple sclerosis. Our purpose was to assess whether different patterns of cortical recruitment are associated with the short term evolution of definite multiple sclerosis (MS). METHODS: We followed for 1 year a group of patients with clinically isolated syndromes (CISs) suggestive of MS and compared the baseline movement-associated patterns of cortical activations between those patients with and those without evolution to definite MS. RESULTS: Those patients in whom MS did not evolve had more significant activations of several areas part of the "classic" motor network; those who went on to develop MS had more significant activations of several regions in the frontal, parietal, temporal, and occipital lobes. CONCLUSION: In CIS patients, the extent of early cortical reorganization following tissue injury might be a factor associated with a different disease evolution. PMID- 15891173 TI - Fast detection of diffuse axonal damage in severe traumatic brain injury: comparison of gradient-recalled echo and turbo proton echo-planar spectroscopic imaging MRI sequences. AB - BACKGROUND AND PURPOSE: Diffuse axonal injury (DAI) is a common type of primary neuronal injury in patients with severe traumatic brain injury (TBI), and is frequently accompanied by tissue tear hemorrhage. T2-weighted gradient-recalled echo (GRE) sequences are more sensitive than T2-weighted spin-echo images for detection of hemorrhage. The purpose of this study is to compare turbo Proton Echo Planar Spectroscopic Imaging (t-PEPSI), an extremely fast sequence, with GRE sequence in the detection of DAI. METHODS: Twenty-one patients (mean age 26.8 years) with severe TBI occurred at least 3 months earlier, underwent a brain MR Imaging study on a 1.5-T scanner. A qualitative evaluation of the t-PEPSI sequences was performed by identifying the optimal echo time and in-plane resolution. The number and size of DAI lesions, as well as the signal intensity contrast ratio (SI CR), were computed for each set of GRE and t-PEPSI images, and divided according to their anatomic location as lobar and/or deep brain. RESULTS: There was no significant difference between GRE and t-PEPSI sequences in the detection of the total number of DAI lesions (291 vs. 230, respectively). GRE sequence delineated a higher number of DAI in the temporal lobe compared to the t PEPSI sequence (74 vs. 37, P < .004), while no differences were found for the other regions. The SI CR was significantly lower with the t-PEPSI than the GRE sequence (P < .00001). CONCLUSION: Owing to its very short scan time and high sensitivity to the hemorrhage foci, the t-PEPSI sequence may be used as an alternative to the GRE to assess brain DAI in severe TBI patients, especially if uncooperative and medically unstable. PMID- 15891174 TI - Unilateral subcortical calcification: a manifestation of dural arteriovenous fistula. AB - We describe the imaging findings in a dural arteriovenous fistula (AVF) with unilateral subcortical calcification. A 50-year-old woman patient suffered from hypertension and chronic headache. Recently, marked headache and a changed consciousness level were noted. The imaging studies demonstrated left subcortical calcification and cerebral sulcus effacement. MR imaging and angiography revealed multiple abnormal tortuous vessels, mainly from left external carotid artery with left-sided transverse sinus occlusion. The final diagnosis was type II a + b dural AVF (classification of Djindjian and Merland), and the patient underwent endovascular embolization and radiosurgery. PMID- 15891175 TI - Tumefactive cysts: a delayed complication following radiosurgery for cerebral arterial venous malformations. AB - We report five cases of symptomatic delayed cyst formation after radiosurgery for intracranial arterial venous malformations. Median time to discovery of the cysts following most recent radiosurgery was 59 months (range, 34-89 months). In all five patients, the radiation therapy-induced cysts exhibited tumefactive characteristics, including vasogenic edema, mass effect, and nodular enhancement. Despite these malignant features, these cystic lesions should be recognized as a benign complication of radiosurgery so that proper treatment (i.e., cystoperitoneal shunt surgery or excision) can be initiated. PMID- 15891176 TI - Radiation-induced cavernomas of the brain. AB - The purpose of this report is to add support to the growing literature that there is a correlation between radiation and cavernomas of the brain, particularly if the radiation is received in childhood, as well as to increase awareness of this correlation in the radiology community. Retrospective review of our experience returned five patients who received radiation therapy while they were children and developed cavernomas in the irradiated tissues 3-41 years later. Cavernomas should be considered in the differential diagnosis of a hemorrhagic lesion in any patient who has received previous CNS radiation, particularly if he or she underwent radiation therapy in childhood. PMID- 15891177 TI - Active hemorrhage into a postresection cavity detected by neuro-CT angiography. AB - We describe a case demonstrating active extravasation of contrast material into a hematoma resection cavity during CT angiography (CTA) that necessitated emergent reexploration, decompression, and hemostatic control. Our case highlights the value of neuro-CTA in the immediate postoperative setting and describes another scenario where CTA has added value. Prompt recognition of contrast extravasation is critical to the diagnosis and ultimately affects the quality of patient care. PMID- 15891178 TI - Bilateral germinoma involving the basal ganglia and cerebral white matter. AB - We describe radiologic findings of germinoma involving bilateral basal ganglia and cerebral white matter. In the early stage, T1-weighted MR images showed hyperintense areas in the bilateral globi pallidi. Later, T2-weighted images showed hyperintense areas involving bilateral white matter. The MR imaging findings reported here could help facilitate the early diagnosis of germ cell tumors in young patients. PMID- 15891179 TI - MR ventriculocisternography by using 3D balanced steady-state free precession imaging: technical note. AB - This study investigated the effects of flip angle setting in 3D balanced steady state free precession (SSFP) imaging on CSF-parenchyma contrast and section aliasing artifacts. Theoretical derivations indicated that the extent of section aliasing artifacts decreased as the flip angle was lowered, at the expense of a sacrifice in CSF-parenchyma contrast. Experimental data agreed closely with theoretical predictions. A flip angle of about 40 degrees is therefore recommended for 3D balanced SSFP MR ventriculocisternography. PMID- 15891180 TI - Selective neck dissection: CT and MR imaging findings. AB - BACKGROUND AND PURPOSE: Selective neck dissection (SND) has become a common surgical procedure for selectively treating known or potential metastatic nodal disease from head and neck cancer while preserving functional structures. The purpose of this article is to describe the expected CT and MR findings after SND. METHODS: CT (26/27) or MR images (1/27) from 27 consecutive patients treated with SND for either staging or nodal control of head and neck malignancy were retrospectively reviewed by two experienced head and neck radiologists. One patient had bilateral SND. The quantity of deep cervical fat was subjectively assessed, as was patency of the ipsilateral internal jugular vein (IJV) and asymmetry in size and contour of the sternocleidomastoid (SCM), trapezius, and infrahyoid strap muscles. The presence of the submandibular gland was noted. RESULTS: Twenty-seven of 28 necks had marked decrease in fat beneath the SCM muscle. This resulted in the muscle directly abutting the paraspinal muscles in most cases. The SCM muscle contour and size was asymmetric or flattened and atrophic in 16/28 necks. Atrophy of the infrahyoid strap muscles was seen in 8/28 necks. Six of 28 had no detectable IJV, and it was presumably thrombosed. Submandibular gland was not present in 17/28 cases. CONCLUSION: The imaging findings after SND are characteristic and reflect the type of surgery performed. If level I nodes are removed, the submandibular gland is absent. Marked decrease in deep cervical fat is common. Changes in and around the SCM muscle are routinely seen and include posterior and medial displacement of the muscle, distortion and flattening of the muscle, or atrophy, despite surgical preservation of spinal accessory nerve. Finally, although the IJV is not resected in SND, nonvisualization of the vein on postoperative images may reflect thrombosis. PMID- 15891181 TI - Perfusion CT for head and neck tumors: pilot study. AB - BACKGROUND AND PURPOSE: Differentiation of malignant from benign head and neck lesions is often very difficult on imaging studies, especially in patients with treated cancer. We evaluated the feasibility and reproducibility of perfusion CT (CTP) after enhanced head and neck CT and attempted to differentiate benign from malignant processes. METHODS: CTP was attempted in 17 patients after head and neck contrast-enhanced CT. Data were postprocessed by using deconvolution-based perfusion analysis. Ipsilateral and contralateral internal, external, and common carotid arteries were used as arterial input vessels. Postprocessing-generated maps showed mean transit time (MTT), blood volume, blood flow, and capillary permeability surface product. Two readers independently placed regions of interest through the primary site, salivary glands, thyroid gland, paraspinous muscles, muscles of mastication, sternocleidomastoid muscle, base of tongue, and subcutaneous fat. One reader repeated the measurements on separate dates. Data were statistically analyzed, and histologic specimens were obtained. RESULTS: CTP was not possible in four patients, and one was lost to follow-up. Of the remaining 12, five had cancer, and seven had benign processes. We found no significant interreader or intrareader differences and no significant difference between various input vessels. Differentiation between malignant and nonmalignant lesions was most reliable by using MTTs. Measurements were comparable to those in the literature. CONCLUSION: CTP after enhanced head and neck CT is feasible, except perhaps at the laryngeal level. It appears to be reader independent and reproducible regardless of the input vessel. CTP shows promise in distinguishing benign and malignant processes, primarily by means of MTTs. PMID- 15891182 TI - Usefulness of the apparent diffusion coefficient in line scan diffusion-weighted imaging for distinguishing between squamous cell carcinomas and malignant lymphomas of the head and neck. AB - BACKGROUND AND PURPOSE: Squamous cell carcinoma (SCC) and lymphoma are common malignant tumors of the head and neck. The purpose of this study was to determine whether the apparent diffusion coefficient (ADC) in line scan diffusion-weighted imaging (LSDWI) is useful for distinguishing between SCC and lymphoma of the head and neck. METHODS: LSDWI was prospectively performed in 39 patients with SCC and in 14 patients with lymphoma. Images were obtained with a diffusion-weighted factor (b factor) of 5 and 1000 s/mm(2), and ADC maps were generated. ADC values were measured for the two types of tumor. RESULTS: Mean ADC values were 0.96 +/- 0.11 x 10(-3) mm(2)/s for SCC and 0.65 +/- 0.09 x 10(-3) mm(2)/s for lymphoma; the difference was significant (P < .001). All but one of the patients with lymphoma had ADC values lower than the lowest ADC (0.76 x 10(-3) mm(2)/s) in patients with SCC. When an ADC of 0.76 x 10(-3) mm(2)/s was used to distinguish between SCC and lymphoma, accuracy was 98% (52 of 53 lesions). CONCLUSION: ADC values appear to be useful for distinguishing between SCC and lymphoma in the head and neck. PMID- 15891183 TI - MR imaging-guided interstitial photodynamic laser therapy for advanced head and neck tumors. AB - Photodynamic therapy (PDT) is a site-specific tumor treatment involving the administration of a photosensitizer activated by the local application of light. In interstitial PDT (IPDT), multiple laser fibers are inserted into the depth of the tumor. Image guidance is essential for accurate, safe, and uniform light delivery. We report a novel technique of IPDT for advanced head and neck tumors involving an open interventional MR system. Initial results are encouraging, with minimal procedural morbidity, successful palliation of symptoms, and prolongation of expected survival time. PMID- 15891184 TI - MR imaging of salivary duct carcinoma. AB - BACKGROUND AND PURPOSE: Salivary duct carcinoma (SDC) is regarded as a high-grade malignancy in the current classification of salivary gland neoplasms. The aim of our study was to describe the MR imaging features of SDC. METHODS: Nine patients with SDC underwent MR imaging study. The apparent diffusion coefficient (ADC) values of SDCs were measured from diffusion-weighted images. Time-signal intensity curves (TICs) of the tumors on dynamic MR images were plotted, and washout ratios were also calculated. TICs were divided into four types: type A, curve peaks <120 seconds after administration of contrast material with high washout ratio (> or =30%); type B, curve peaks <120 seconds with low washout ratio (<30%); type C, curve peaks >120 seconds; type D, nonenhanced. We correlated the MR findings of SDC with the pathologic findings. RESULTS: All tumors had ill-defined margins and showed low to moderately high signal intensity for contralateral parotid gland on T2-weighted images. The average of the ADC values of the SDCs was 1.16 +/- 0.14 [SD] x 10(-3)mm(2)/s. Seven of nine (78%) tumors had type B enhancement. On the other hand, six of nine (67%) tumors with rich fibrotic tissue also had type C enhancement. CONCLUSION: The findings of ill defined margin, early enhancement with low washout ratio (type B), and low ADC value (1.22 x 10(-3)mm(2)/s) were useful for suggesting malignant salivary gland tumors. Although it was reported that type C enhancement was specific for pleomorphic adenoma, SDC frequently has type C-enhanced focus. PMID- 15891185 TI - MR microscopy of the parotid glands in patients with Sjogren's syndrome: quantitative MR diagnostic criteria. AB - BACKGROUND AND PURPOSE: MR imaging of the salivary glands has been applied to the diagnosis of Sjogren's syndrome; however, the diagnosis remains qualitative. We sought to establish and evaluate quantitative MR imaging criteria for the diagnosis of Sjogren's syndrome. METHODS: MR imaging with a 47-mm microscopy coil was performed in 83 patients with xerostomia (55 patients with Sjogren's syndrome, 28 without Sjogren's syndrome). MR images were obtained by T1-weighted and fat-suppressed T2-weighted imaging and by MR sialography of the parotid glands. MR imaging findings of the parotid glands in Sjogren's syndome included increases in fat areas and decreases in intact lobule areas. These MR images were morphometrically analyzed for the diagnostic criteria. RESULTS: MR imaging with a microscopy coil demonstrated well the details of the damaged parotid glands in patients with xerostomia. Quantitative MR imaging of fat, intact gland lobule, and number of sialoectatic foci significantly and highly correlated with severity of disease. Receiver operating characteristic (ROC) curve analysis demonstrated that quantitative MR imaging yielded high diagnostic ability in differentiating patients with xerostomia who have Sjogren's syndrome from those without Sjogren's syndrome, with areas under the ROC curve of 0.94 for fat area, 0.98 for intact lobule area, and 0.91 for number of sialoectatic foci. The best cutoff points by quantitative MR imaging were each associated with high sensitivity and specificity, and, when used in combination, yielded 96% sensitivity and 100% specificity. CONCLUSION: Quantitative MR imaging effectively differentiated the parotid glands in patients with xerostomia who have Sjogren's syndrome from those without the syndrome and provided criteria for staging the gland disease. PMID- 15891186 TI - Preoperative imaging of thyroid goiter: how imaging technique can influence anatomic appearance and create a potential for inaccurate interpretation. AB - The radiologist needs to be aware of a potential pitfall that can influence the imaging appearance of thyroid goiter. Whether the patient is imaged with the arms overhead or by the side may affect the apparent mediastinal excursion of a goiter. CT scans obtained with the patient's arms by the side are more accurate for determining substernal extent of goiter than when the arms are overhead, a position usually used in chest CT. Ultimately, this difference in imaging technique may have a profound effect on the adjacent structures impacted by the goiter and may influence the planned surgical approach. PMID- 15891187 TI - Barotrauma presenting as temporal lobe injury secondary to temporal bone rupture. AB - We present the case of a scuba diver who experienced acute ear pain during ascent from a dive. CT imaging was performed because of severe unrelenting headache. Blood and gas was identified within the epidural space of the middle fossa and over the petrous bone, as well as hemorrhage within the adjacent temporal lobe. To the best of our knowledge, this is the first case of CT-documented barotrauma affecting the brain as a result of middle-ear gas rupturing through the tegmen tympani. PMID- 15891188 TI - Nocardial endophthalmitis and subretinal abscess: CT and MR imaging features with pathologic correlation: a case report. AB - Ocular nocardiosis is a rare but potentially aggressive process. Clinically, it can mimic other disease entities, including neoplasia and other types of infection. We present a case of nocardial panophthalmitis progressing to subretinal abscess and emphasize the radiologic and clinical features. PMID- 15891189 TI - Self-expandable stent-assisted coiling of wide-necked intracranial aneurysms: a single-center experience. AB - BACKGROUND AND PURPOSE: Endovascular treatment of wide-necked aneurysms remains a therapeutic challenge. We conducted this study to evaluate the angiographic results and clinical outcome of patients treated with stent-assisted coiling by using a recently available self-expandable intracranial stent. METHODS: A retrospective review of all patients treated with self-expandable stent-assisted coiling between September 2002 and December 2003 was done. Treatment was attempted in 32 patients with 35 aneurysms. Four of the aneurysms were ruptured. All had either a dome-to-neck ratio less than 2 and/or a neck diameter of 5 mm or larger. Following stent placement, coiling was attempted in 33 of 34 aneurysms. The technical success of the procedure, procedure related complications, and the angiographic results were documented. RESULTS: In 34 of 35 aneurysms, stent deployment across the neck of the aneurysm was successful. Coiling was performed successfully in 30 of 33 aneurysms. In 20 aneurysms, immediate posttreatment angiography showed either total (17%) or satisfactory (50%) occlusion. Procedure related mortality occurred in one patient (3.1%). Adverse events occurred in eight patients (25%); in three of them permanent neurologic deficit resulted (9.3%). In six patients, thrombus formation occurred within the stented segments during the procedure and reopro infusion was used. Follow-up angiography was available in 12 (40%) of 30 treated aneurysms. CONCLUSION: In our practice use of the self-expandable stent seemed to facilitate endovascular treatment of wide necked intracranial aneurysms. Difficulty of deployment and stent thrombogenicity are the main drawbacks of the system. PMID- 15891190 TI - Aneurysmal pressure changes with nondetachable balloon placement and fluid infusion: rationale for liquid embolization. AB - BACKGROUND AND PURPOSE: To improve the safety and efficacy of liquid embolization, we evaluated changes in pressures in experimental aneurysms. METHODS: We created three replicas of a lateral sidewall aneurysm and placed them in a physiologic flow circuit. A 3 x 10-mm nondetachable balloon was positioned in the parent vessel across the aneurysmal neck. Intra-aneurysmal pressures were measured at baseline and after balloon inflation. Fluid was infused into the sac via a 1.45F microcatheter during inflation, and maximal pressures were noted. Measurements were repeated eight times in each aneurysm. RESULTS: After balloon inflation, average intra-aneurysmal pressures increased: 12 mm Hg (13%, sigma(n - 1) = 0.46) for aneurysm 1 (baseline mean arterial pressure [MAP], 94 mm Hg), 15 mm Hg (58%, sigma(n - 1) = 0.88) for aneurysm 2 (baseline MAP, 26 mm Hg), and 15 mm Hg (58%, sigma(n - 1) = 0.92) for aneurysm 3 (baseline MAP, 26 mm Hg). During inflation and infusion, pressures increased slightly: 1.1 (0.94%, sigma(n - 1) = 0.64), 1.6 (3.9%, sigma(n - 1) = 1.1), and 1.9 (4.6%, sigma(n - 1) = 1.2) mm Hg for aneurysms 1, 2, and 3, respectively. Despite complete balloon occlusion of the distal aneurysmal neck, a channel between the proximal aneurysmal neck and the parent-vessel lumen persisted along the microcatheter. Fluid exited the sac via this channel, preventing a concomitant, significant increase in pressure during infusion. CONCLUSION: Intra-aneurysmal pressure modestly increased with inflation of a parent-vessel balloon across the neck. When liquid was infused into the sac during inflation, further increases were minimal. PMID- 15891191 TI - Treatment options for wide-necked intracranial aneurysms using a self-expandable hydrophilic coil and a self-expandable stent combination. AB - The endovascular treatment of wide-necked intracranial aneurysms is evolving. We report our initial experience in treating seven wide-neck intracranial aneurysms by using a new self-expanding stent, the Neuroform, in combination with a new generation of coils that incorporate a self-expandable hydrophilic lining, the HydroCoil. PMID- 15891192 TI - Efficacy of treatment of severe carotid bifurcation stenosis by using self expanding stents without deliberate use of angioplasty balloons. AB - BACKGROUND AND PURPOSE: One of the limitations of carotid artery angioplasty is the potential for embolic stroke. Our purpose was to assess whether the force of a self-expanding stent alone is usually sufficient to dilate severely stenotic atherosclerotic plaques without the deliberate use of an angioplasty balloon. If so, the procedural stroke risk might be reduced. METHODS: Over a 30-month period, 21 consecutive patients were prospectively identified with severe symptomatic carotid artery stenosis (>70% NASCET [North American Symptomatic Carotid Endarterectomy Trial]) and relative indications for endovascular treatment. All underwent treatment with the aim of deploying a self-expanding stent across the stenosis without the use of angioplasty balloons or distal protection devices. RESULTS: Stent deployment was successful in 20/21 patients. In one patient, the stent could not be deployed without balloon predilatation and a stroke occurred. In the other 20 patients, angiography before and immediately after stent deployment showed a reduction in the mean stenosis from 83% to 49%. A second periprocedural stroke occurred as a result of early stent thrombosis at 4 days in a patient who stopped Acetylsalicylic acid while undergoing bowel preparation for colon surgery. He made a good recovery. Average duration of follow-up imaging was 19 months (range, 1-44 months). During the follow-up period there were four deaths, all unrelated to the carotid disease, and no major strokes. At 5-11 months, the average residual stenosis was 21%, which remained stable in 16 of the 18 patients studied between 12 and 44 months (average, 24 months). At last follow up, in these 16 the mean peak systolic velocity was 123 cm/s (range, 60-238 cm/s) and the mean internal-to-common carotid ratio was 1.8. The other two patients were long-term failures of the "stent-only" approach. In one patient, a heavily calcified plaque prevented stent expansion and the artery occluded at 18 months with a minor stroke. In the second patient, a recurrent stenosis developed at 16 months with one episode of transient ischemic attack. CONCLUSION: Deployment of a self-expanding stent alone resulted in a favorable and more gradual reduction of severe symptomatic carotid stenosis. Improvements in stent profile and chronic outward force may widen the indications for simple stent placement without the use of balloons or adjunctive protection devices. PMID- 15891193 TI - Emergency carotid artery stent placement in patients with acute ischemic stroke. AB - BACKGROUND AND PURPOSE: An effective intervention has not yet been established for patients with acute ischemic stroke who present with serious neurologic symptoms due to occlusion or a high-grade stenosis of the internal carotid artery (ICA). The aim of our retrospective study was to investigate the feasibility, safety, and efficacy of emergency carotid artery stent placement to improve neurologic symptoms and clinical outcome. METHODS: Of 896 consecutive patients with acute ischemic stroke who were admitted to our institution within 7 days of onset from July 2000 to June 2003, 17 patients (1.9%) with occlusion or a high grade stenosis of the ICA underwent emergency carotid artery stent placement. We reviewed their records for neurologic outcome, per the National Institutes of Health Stroke Scale (NIHSS) score, before and at 7 days after stent placement; clinical outcome, per the modified Rankin Scale score (mRS), at 90 days; frequency of procedure-related complications within 30 days; and recurrence rate of ipsilateral ischemic stroke within 90 days. RESULTS: Carotid lesions were dilated completely in all patients. Median NIHSS scores before emergency stent placement and at 7 days were 12 and 5, respectively, showing significant improvement (P < .01, Wilcoxon rank sum test). Ten patients (59%) had favorable outcomes (mRS score 0-1) at 90 days. Irreversible complications occurred in two patients (12%): distal embolism in one and intracerebral hemorrhage in the other. No ipsilateral ischemic stroke recurred. CONCLUSION: Emergency carotid artery stent placement can improve the 7-day neurologic outcome and may improve the 90 day clinical outcome in selected patients with ischemic stroke. PMID- 15891194 TI - Cerebral arteriovenous fistulas induced by dural arteriovenous shunts. AB - Dural arteriovenous shunts and pial arteriovenous fistulas are uncommonly associated. Their etiology, pathogenesis, and natural history are still unclear and are likely different. We present three cases of high-flow dural arteriovenous shunts associated with pial arteriovenous fistulas and discuss their pathogenesis, anatomic association, and angioarchitecture. We propose that venous steal effect in the dural sinus secondary to the high-flow dural arteriovenous shunt induced the pial arteriovenous fistulas. Treatment of the high-flow dural arteriovenous shunts and the induced pial arteriovenous fistulas are discussed. PMID- 15891195 TI - Prevalence of leukoencephalopathy in children treated for acute lymphoblastic leukemia with high-dose methotrexate. AB - BACKGROUND AND PURPOSE: An effective treatment for acute lymphoblastic leukemia (ALL), intravenous (IV) methotrexate (MTX) has a notable toxic effect on the CNS, with leukoencephalopathy (LE) being the most common form. The purpose of this study was to use objective quantitative MR imaging to prospectively assess potential risk factors on the temporal evolution of LE in patients treated for ALL. METHODS: We evaluated the longitudinal prevalence of LE in 45 children treated for ALL in a single institutional protocol including seven courses of IV MTX and no cranial irradiation. Differences in signal intensity on T2-weighted and fluid-attenuated inversion recovery (FLAIR) images between hyperintense regions and normal-appearing genu were used to quantitatively detect LE. Cox proportional regression was used to estimate the effect of covariates (e.g., sex, MTX dose, age at diagnosis) on the prevalence of LE. After influential factors were identified, a generalized linear model was determined to predict the probability of LE in new patients. The model was necessary to facilitate statistical testing between examinations. RESULTS: Increasing exposure, which corresponding to more courses and higher doses of IV MTX, influenced the prevalence of LE. The prevalence of LE was significant reduced approximately 1.5 years after the completion of IV MTX. CONCLUSION: Higher doses and more courses of IV MTX placed patients at a higher risk for LE; many of the changes resolved after the completion of therapy. The effect of these changes on neurocognitive functioning and quality of life in survivors remains to be determined. PMID- 15891196 TI - Hippocampal alterations in children with temporal lobe epilepsy with or without a history of febrile convulsions: evaluations with MR volumetry and proton MR spectroscopy. AB - BACKGROUND AND PURPOSE: The causal effect of early febrile convulsions (FC) on later-onset temporal lobe epilepsy (TLE) remains unclear. In this study, we sought to examine the hippocampal alterations in epileptic children with or without FC history by using MR spectroscopy and volumetry. METHODS: Fifty-five children ranging in age from 18 months to 15 years were enrolled in this study. Subjects were divided into three groups: the control group without either TLE or history of FC (n = 16), the TLE group with early history of FC (TLE + FC; n = 22), and the TLE group without FC history (n = 17). Measurement of hippocampal volume (HV) was performed on thin section T1-weighted images acquired with a 3D gradient echo MR image and normalized by the intracranial volume. Each individual subject had two measures of lateralization; one gives the smaller side of HV and the other the contralateral larger side of HV, assuming that the side with smaller HV is the possible primary site of seizure focus and the contralateral larger HV the secondary or normal site. Single-voxel proton MR spectroscopy of the hippocampi was performed, with metabolic ratio n-acetylaspartate (NAA)/choline (Cho) + creatine plus phosphocreatine (Cr) calculated and grouped separately as were with volumetry. RESULTS: The overall mean HV for the control group was 2.61 +/- 0.21 cm(3) at an average intracranial volume of 965 +/- 241 cm(3), and the asymmetry index for hippocampal volume was (2.32 +/- 1.58)%. The overall mean HV was 2.30 +/- 0.33 cm(3) for TLE + FC group and 2.34 +/- 0.33 cm(3) for TLE group. Mean HV differed significantly for the three groups (P < .01). When the small and large sides were analyzed separately, significant differences were found between control and TLE as well as between control and TLE + FC for the smaller side (P < .05), whereas for the larger side significant differences were found only between control and TLE + FC. In MR spectroscopic measurements, the mean NAA/(Cr + Cho) of bilateral hippocampi was 0.77 +/- 0.06 for control group, 0.62 +/- 0.12 for TLE + FC group, and 0.66 +/- 0.11 for TLE group. In terms of statistically significant difference between groups, spectroscopic results were similar to volumetric measurements, except that there was no significant interaction effect between groups and measures of asymmetrical indices (P = .272). CONCLUSION: Children with TLE and early history of FC tend to have lower hippocampal volumes and NAA/(Cr + Cho) ratios than do TLE children without FC history. The TLE + FC group seems to have increased vulnerability of the contralateral hippocampus as compared with TLE group. MR volumetry and spectroscopy are equally capable of showing the trends of hippocampal alternations in children with TLE with or without FC history. PMID- 15891197 TI - Proton MR spectroscopic imaging depicts diffuse axonal injury in children with traumatic brain injury. AB - BACKGROUND AND PURPOSE: Diffuse axonal injury (DAI) after traumatic brain injury (TBI) is important in patient assessment and prognosis, yet they are underestimated with conventional imaging techniques. We used MR spectroscopic imaging (MRSI) to detect DAI and determine whether metabolite ratios are accurate in predicting long-term outcomes and to examine regional differences in injury between children with TBI and control subjects. METHODS: Forty children with TBI underwent transverse proton MRSI through the level of the corpus callosum within 1-16 days after injury. T2-weighted, fluid-attenuated inversion recovery, and susceptibility-weighted MR imaging was used to identify voxels as normal appearing or as nonhemorrhagic or hemorrhagic injury. Neurologic outcome was evaluated at 6-12 months after injury. Metabolite ratios for total (all voxels), normal-appearing, and hemorrhagic brain were compared and used in a logistic regression model to predict long-term outcome. Total and regional metabolite ratios were compared with control data. RESULTS: A significant decrease in N acetylaspartate (NAA)/creatine (Cr) and increase in choline (Cho)/Cr (evidence of DAI) was observed in normal-appearing (P < .05) and visibly injured (hemorrhagic) brain (P < .001) compared with controls. In normal-appearing brain NAA/Cr decreased more in patients with poor outcomes (1.32 +/- 0.54) than in those with good outcomes (1.61 +/- 0.50, P = .01) or control subjects (1.86 +/- 0.1, P = .00). In visibly injured brains, ratios were similarly altered in all patients. In predicting outcomes, ratios from normal-appearing and visibly-injured brain were 85% and 67% accurate, respectively. CONCLUSION: MRSI can depict injury in brain that appears normal on imaging and is useful for predicting long-term outcomes. PMID- 15891198 TI - Detection of cerebral {beta}-hydroxy butyrate, acetoacetate, and lactate on proton MR spectroscopy in children with diabetic ketoacidosis. AB - BACKGROUND AND PURPOSE: Ketone bodies provide important alternate fuel for brain metabolism, and their transport into the brain increases with prolonged fasting. During diabetic ketoacidosis (DKA), serum ketone concentrations markedly increase; however, little is known about whether ketone bodies accumulate in cerebral tissues during DKA. We used proton MR spectroscopy (MRS) to detect cerebral beta-hydroxy butyrate (betaOHB) and acetone/acetocaetate (AcAc) in children with DKA. METHODS: Twenty-five children underwent brain MRS: nine within 4 hours of the start of treatment for DKA; 11, at 4-8 hours; and five, at 8-12 hours. MRS was repeated after their recovery from the DKA episode at > or =72 hours after the start of treatment. MRS was evaluated for peaks corresponding to betaOHB (doublet centered on 1.20 ppm) and lactate (doublet centered on 1.33 ppm). Difference spectroscopy was used to identify the AcAc peak at 2.22-2.26 ppm. RESULTS: betaOHB was detected in 13 children (52%), more frequently within 4 hours (eight children, 89%) than after 4 hours (five children, 31%). AcAc was detected in 15 children (60%), more frequently at >4 hours after the start of treatment (12 patients, 75%) than in the first 4 hours (three patients, 33%). Lactate was detected in five children (18%), all within the first 8 hours of treatment. CONCLUSION: In children, betaOHB and AcAc accumulate in the brain during DKA, and they can be detected on MRS. Care should be taken in interpreting MRS results in patients with DKA to avoid erroneously attributing betaOHB peaks to lactate. PMID- 15891199 TI - Diffusion-weighted monitoring of conservatively treated pyogenic brain abscesses: a marker for antibacterial treatment efficacy. PMID- 15891200 TI - Conventional t1-weighted imaging in the diagnosis and assessment of brain abscesses. PMID- 15891201 TI - The promises and challenges of rigorous research. PMID- 15891203 TI - St John's for depression, worts and all. PMID- 15891206 TI - Antioxidants for children with kwashiorkor. PMID- 15891207 TI - The fetal origins hypothesis--10 years on. PMID- 15891208 TI - Open access, impact, and demand. PMID- 15891209 TI - Monitoring surgical mortality. PMID- 15891210 TI - Global functions at the World Health Organization. PMID- 15891214 TI - NICE says that patients' age should affect treatment. PMID- 15891212 TI - Half of patients in intensive care receive suboptimal care. PMID- 15891216 TI - English surgeons may at last be about to become doctors. PMID- 15891217 TI - Medical societies react against public access to findings. PMID- 15891218 TI - UK and US groups announce new initiatives on open access publishing. PMID- 15891221 TI - Medical students should be added to GMC register, says judge. PMID- 15891219 TI - Doctors accuse India of massaging leprosy figures. PMID- 15891222 TI - More pubs will escape smoking ban than UK government has claimed. PMID- 15891224 TI - Doctor was "near to breaking point" when she exaggerated abuse. PMID- 15891225 TI - New law aims to distance the FDA from the drug industry. PMID- 15891226 TI - WHO's attempts to eradicate polio are thwarted in Africa and Asia. PMID- 15891227 TI - Mumps outbreaks across England and Wales in 2004: observational study. PMID- 15891228 TI - Hyperhidrosis. PMID- 15891229 TI - Mumps and the UK epidemic 2005. PMID- 15891230 TI - Major trauma. PMID- 15891231 TI - Building a framework for trust: critical event analysis of deaths in surgical care. PMID- 15891232 TI - Excellent review scheme for critical incidents but insufficient for revalidation. PMID- 15891233 TI - Failure to act on good intentions. PMID- 15891234 TI - Revalidation in the UK. PMID- 15891235 TI - Do selective serotonin reuptake inhibitors cause suicide? Figures look doubtful. PMID- 15891236 TI - Do selective serotonin reuptake inhibitors cause suicide? Risk of suicide should be assessed for whole class of antidepressants. PMID- 15891237 TI - Do selective serotonin reuptake inhibitors cause suicide? Data seem to be incorrect. PMID- 15891238 TI - Do selective serotonin reuptake inhibitors cause suicide? Let's keep it in perspective. PMID- 15891239 TI - Do selective serotonin reuptake inhibitors cause suicide? Discrediting old drugs may be useful in marketing new ones. PMID- 15891240 TI - Do selective serotonin reuptake inhibitors cause suicide? Suicide rate of 15% in editorial is misleading. PMID- 15891241 TI - Many patients may not understand consent forms. PMID- 15891242 TI - Attributable lung cancer risk from radon in homes may be low. PMID- 15891243 TI - Do selective serotonin reuptake inhibitors cause suicide? Antidepressant prescribing to children and adolescents by GPs has fallen since CSM advice. PMID- 15891244 TI - Over-the-counter statin therapy--pros and cons. PMID- 15891245 TI - New diagnosis of left atrial myxoma in a 93-year-old woman. AB - A 93-year-old woman with a history of hypertension was noted to have a mass in the left atrium prolapsing partially through the mitral valve on a routine surface echocardiogram in 2002. A transesophageal echocardiogram was then performed revealing an irregular mass with an appearance of protruding fronds of tissue. The echogenicity of the mass was inhomogeneous with some areas of punctate calcification, and the mass was attached to the interatrial septum. The echocardiographic appearance of the mass was almost pathognomic of atrial myxoma, and this is the oldest patient yet to have been diagnosed with a probable myxoma. The patient did not want surgical removal of the mass. She is still asymptomatic at the age of 95 years. PMID- 15891246 TI - Continued use of nonsteroidal topical treatments: comment on the FDA talk paper. PMID- 15891248 TI - Self-mutilation: an addictive, contagious, and easily spotted cry for help. PMID- 15891249 TI - Combination therapy with adapalene gel 0.1% and doxycycline for severe acne vulgaris: a multicenter, investigator-blind, randomized, controlled study. AB - BACKGROUND: Combination therapy with a topical retinoid and an antibiotic is recognized as a rational and effective approach for the treatment of acne vulgaris. Adapalene, a naphthoic acid derivative with anti-inflammatory and receptor-selective retinoid properties, is safe and well tolerated. While the combination of adapalene with oral or topical antibiotics has been shown to deliver a superior and faster response than an antibiotic alone, the clinical benefits of a combination of adapalene and doxycycline, the most frequently prescribed oral antibiotic for acne in the United States, have yet to be evaluated. OBJECTIVE AND METHODS: In a 12-week study, the efficacy and safety of the combination of adapalene gel 0.1% with doxycycline was compared with doxycycline alone for the treatment of severe acne. Subjects were randomized to receive doxycycline once daily in the morning and either adapalene or vehicle once daily in the evening. RESULTS: At Week 12, the combination adapalene doxycycline was significantly superior to doxycycline alone for change from baseline in total (p<0.001), inflammatory (p=0.02), and noninflammatory (p<0.001) lesions. Significant differences in total lesions were observed as early as Week 4 (p=0.04). Both treatments were well tolerated, and no serious adverse events were reported. CONCLUSIONS: The study demonstrates that the combination of adapalene and an oral antibiotic provides a superior and faster benefit than antibiotic therapy alone and should be considered at the initiation of treatment. PMID- 15891250 TI - Pimecrolimus for treatment of acne rosacea. AB - BACKGROUND: Rosacea is a common disease, which is often resistant to treatment. Topical calcineurin inhibitors have been efficacious in the treatment of other inflammatory disorders of the skin, and tacrolimus has been reported as an effective treatment option for erythrotelangiectatic rosacea. OBJECTIVE: Because of the benefits seen with tacrolimus in previous publications, we investigated the efficacy of a closely related compound, pimecrolimus, in patients with erythrotelangiectatic, papulopustular, and edematous rosacea. METHODS: Twelve patients with erthryotelangiectatic or papulopustular rosacea who had failed conventional therapy were treated with topical pimecrolimus cream twice daily for 12-18 weeks. No patients had used any other treatment for rosacea within 30 days of the start of therapy. During the course of the study, no other topical or systemic treatment of rosacea was allowed. RESULTS: Ten of 12 patients showed substantial improvement of erythema, while five of six patients with a papulopustular component noted at least an 80% decrease in the number of lesions. CONCLUSIONS: It appears pimecrolimus may be efficacious in the treatment of erythrotelangiectatic and papulopustular rosacea and may be considered in patients with recalcitrant disease. PMID- 15891251 TI - Lupus miliaris disseminatus faciei. Part I: Significance of histopathologic undertones in diagnosis. AB - BACKGROUND: Since its clinical discovery, lupus miliaris disseminatus faciei has sporadically been reported to have different modes of clinicopathologic expression. OBJECTIVE: The purpose of this study was to work up a list of histopathologic undertones and to project and propagate lupus miliaris disseminatus faciei as an exclusive entity. An upcoming Part II of this study will present an overview of the disease. METHODS: All patients visiting the outpatient clinic conforming to hitherto accepted clinical features were included to study patients' age and sex, duration of the disease, and above all, conduct a detailed histopathology review. Using the detailed information obtained, an endeavor was made to organize the disease into early, fully developed, and late lesions. RESULTS: The details of the various parameters were of great help in evolving this modus operandi. The clinical as well as histopathologic features of lupus miliaris disseminatus faciei are fairly distinct and facilitate visualizing the entity as a spectrum comprising early, fully developed, and late lesions. Further studies are called for. PMID- 15891252 TI - Dercum's disease. AB - Dercum's disease (adiposis dolorosa, lipomatosis dolorosa morbus Dercum), is a rare disorder resulting in painful fatty deposits around the upper legs, trunk, and upper arms. The portrait painted of Dercum's disease is very complicated, with many other disorders seen associated with the disease. There are no clear pathological mechanisms known, although it is suspected that there is either a metabolic or autoimmune component involved. Here, the authors review the literature to date, including some information from their own studies. In particular, the authors will look at the different strands of evidence pointing to the pathological mechanism of the disorder. PMID- 15891253 TI - Clinical management of skin scarring. AB - Not all scars are the same. They vary from normal skin scars to distinct types of abnormal scars, such as hypertrophic and disfiguring keloids. Physicians need to identify different types of skin scars and treat them appropriately. Misdiagnosis and mismanagement of scars can be costly for both the patient and physician. Patients with scars often face aesthetic, physical, psychological, and social consequences that result in substantial emotional and financial costs. This article reviews the spectrum of abnormal scar types and problems associated with scarring. PMID- 15891254 TI - Smoking and psoriasis. AB - Many early studies examining the relationship between tobacco and the development of plaque-type psoriasis suggested a significant positive correlation; however, the majority of these initial studies failed to control for alcohol consumption and presented inconsistent results. The objective of this manuscript is to perform a literature review of articles assessing the relationship between smoking and psoriasis while controlling for confounders such as alcohol consumption. Alcohol-controlled studies suggest that women who are smokers have an up to 3.3-fold increased risk of developing plaque-type psoriasis. Men who are smokers do not exhibit such an increased risk, but studies have shown that smoking more than 10 cigarettes per day by men who are psoriasis patients may be associated with a more severe expression of disease in their extremities. In addition, smoking among both men and women who are psoriasis patients has been shown to reduce improvement rates. These data demonstrate the importance of discouraging smoking, particularly among psoriasis patients. PMID- 15891255 TI - Condyloma with lichen nitidus. AB - A 32-year-old black man presented with two separate eruptions on his penis (Figure). He has had a 10-year history of asymptomatic 1-3 mm shiny papules on the shaft of his penis. He also has had one asymptomatic verrucous brown plaque 1.5 cm in diameter on his penis. A small shave biopsy was performed for each process. The former showed lymphocytes and histocytes in the papillary dermis in a "ball-in-claw" relation to the rete ridges of the epidermis. The latter showed an acanthotic epidermis with papillomatosis. Diagnoses of lichen nitidus and condyloma acuminata were respectively made. As the shiny papules did not bother the patient, but he wanted to be rid of the verrucous plaque, only the latter was treated. Liquid nitrogen was applied to the plaque. The patient was also given podofilox 0.5% gel to apply to the condyloma overnight on Monday, Tuesday, and Wednesday, with a 4-day respite. This cycle was to be repeated for 3 more weeks. He was examined several more times and treated with cryotherapy and podofilox gel with the near disappearance of the condyloma over 4 months. PMID- 15891256 TI - With this eruption, there is not a second to lues. AB - A 28-year-old white man presented to the Emergency Department with a 24-hour history of an eruption on his extremities, trunk, and face. The patient was known to be HIV positive with a CD4 count of 527 and a viral load of 20,300. He denied fever, chills, malaise, and headache. His social history was significant for the fact that he was in a monogamous homosexual relationship. He had no recent travel, pet exposures, or sick contacts. Physical examination revealed stable vital signs and no documented fever. A maculopapular eruption was present on his face, trunk, and extremities (Figures 1 and 2). There was no palmar or plantar involvement. He was treated with diphenhydramine and topical 2.5% hydrocortisone and advised to return if his condition did not improve. Twelve days after the initial evaluation, the patient consulted us again due to progression of his dermatitis. He had no additional complaints other than an eruption on both palms but neither sole. (Figure 3). The eruption now demonstrated erythematous pink-red oval macules and papules 1-2 cm in size distributed on his scalp, face, trunk, and arms. A few papules contained fine collarettes of scale. Further questioning revealed that the patient had experienced a tender rectal ulcer 2 months previously. A punch biopsy and rapid plasma reagin were performed. The histopathologic examination revealed interface dermatitis with lymphocytes, plasma cells, occasional neutrophils, and a prominent lymphoplasmacytic perivascular dermatitis with infiltration of the vessel walls. Warthrin-Starry and Steiner methods demonstrated spirochetes at the dermal-epidermal junction and in vessel walls, consistent with Treponema pallidum (Figure 4). Rapid plasma reagin and fluorescent Treponema antibody were both reactive with a Venereal Disease Research Laboratory (VDRL) of 1:16. The patient was diagnosed as having secondary syphilis and treated with 2.4 million units of IM benzathine penicillin for 3 weeks. His eruption resolved after the initial treatment and he did not experience a Jarisch-Herxheimer reaction. PMID- 15891257 TI - Anatomy of a skin cleanser. PMID- 15891258 TI - Case study: inoculation herpes barbae. AB - A 21-year-old white man in otherwise excellent general health was referred for a painful, progressive, facial eruption with associated fever, malaise, and cervicofacial lymphadenopathy. The patient reported that a vesicular eruption progressed from the left side of his face to also involve the right side of his face over the 48 hours preceding his clinic visit. He also reported some lesions in his throat and the back of his mouth causing pain and difficulty swallowing. Four to 7 days before presentation to us, the patient noted exposure to his girlfriend's cold sore. Additionally, he complained of a personal history of cold sores, but had no recent outbreaks. Physical examination revealed a somewhat ill man with numerous vesicles and donut-shaped, 2-4 mm, crusted erosions predominantly on the left side of the bearded facial skin. There were fewer, but similar-appearing lesions, on the right-bearded skin. The lesions appeared folliculocentric (Figure). Cervical and submandibular lymphadenopathy was present. Oral exam showed shallow erosions on the tonsillar pillars and soft palate. Genital examination was normal. The remainder of the physical exam was unremarkable. A Tzanck smear of vesicular lesions was positive for balloon cells and many multinucleated giant cells with nuclear molding. A viral culture was performed which, in several days, came back positive for herpes simplex virus. The complete blood cell count documented a white blood cell count of 8000/mm3 with 82.6% neutrophils and 9.0% lymphocytes. Based on the clinical presentation and the positive Tzanck smear, the patient was diagnosed with herpes simplex barbae, most likely spread by shaving. The patient was started on acyclovir 200 mg p.o. five times daily for 10 days. Oxycodone 5 mg in addition to acetaminophen 325 mg (Percocet; Endo Pharmaceuticals, Chadds Ford, PA) was prescribed for pain relief. A 1:1:1 suspension of viscous lidocaine (Xylocaine; AstraZeneca Pharmaceuticals LP, Wilmington, DE), diphenhydramine (Benadryl; Pfizer Inc., New York, NY), and attapulgite (Kaopectate; Pfizer Inc., New York, NY) was given as a swish and spit to relieve the oral discomfort. Good hygiene, no skin-to-skin contact with others, and no further shaving to prevent autoinoculation were stressed. He was advised to discard his old razor. PMID- 15891259 TI - Case study: periodic follow-up is necessary in morphea profunda to identify systemic evolution. AB - The term "morphea" includes a wide spectrum of clinical entities, varying from localized plaques of only cosmetic importance to deep lesions resulting in considerable morbidity for the patient. In fact, although survival rates are no different from that of the general population, localized scleroderma may be associated with development of substantial disability, as occurs in deep morphea and in pediatric patients (disabling pansclerotic morphea of children). We report a case of morphea profunda affecting a young man with severe, rapidly progressive, widespread skin involvement and focus on the eventual systemic evolution of such cases. A 40-year-old man was admitted in 2002 for progressive subcutaneous indurations, preferentially involving the right side of the trunk. His health was altogether good, with the exception of a beginning chronic obstructive bronchopneumopathy. There was no family or personal history of dysmetabolic, cardiovascular, neoplastic, or cutaneous disease. Three years earlier, the patient had noted the appearance of two infiltrated, intensely red lesions on the right laterocervical and paraumbilical regions. These had been interpreted as subcutaneous lipomatosis on the basis of an ultrasound scan. The lesions had become progressively larger, while their surface had assumed a scleroatrophic appearance. Thereafter, other lesions had developed on his chest and lower limbs, mostly distributed on the right side of the body. Clinical examination revealed well demarcated, depressed sclerotic plaques with ivory colored centers and erythematous borders ("lilac ring") localized on the neck, chest, and lower abdomen and limbs (Figure 1). They were bound to the deeper structures and arranged in a band-like linear distribution on the right side of the chest and abdomen where they extended horizontally for more than 10 cm in diameter. These lesions were totally asymptomatic. In addition, arborizing telangiectasias were evident on the neck and upper chest (Figure 2). Laboratory investigations provided normal range of erythrocyte sedimentation rat and C reactive protein levels and other inflammation markers. Antinuclear antibody, antidouble-strand DNA, antimitochondrial, anti-extractable antigens (anti centromere, anti-Scl-70, anti-U1RNP), and anti-Borrelia burgdorferi antibodies were negative. Circulating immunocomplexes binding C1q were substantially increased. Oesophageal x-rays and lower limb electromyography were within normal limits; ventilatory function testing revealed a mild obstruction consistent with the beginning of chronic obstructive pulmonary disease. Although nailfold capillaroscopy documented nonspecific findings of connective tissue disease (mega capillaries, segmentary dilatation and destruction), the laser-Doppler flussimetry revealed few signs of microcirculatory abnormalities, in absence of Raynaud's phenomenon. An abdominal wall ultrasonography, performed on a sclerotic plaque, documented thinning of the subcutaneous tissue, with increase of the fibrous component and lower fascia and muscle retraction. The biopsy specimen from the abdominal region included fascia and the subcutaneous tissue (previously obtained from the lower abdomen) with epidermal atrophy, a thickening and homogenization of collagen bundles in the deep dermis and hair reduction. A perivascular lympho-monocytic and plasmacellular infiltration with a dermo epidermal distribution was present. Moreover, septal fibrosis with a perivascular lymphoplasmacellular inflammatory infiltrate was documented within the abdominal rectus muscle. The diagnosis of morphea profunda was made on the basis of clinical and histopathological findings. A therapeutic regimen based on amino benzoic potassium (Potaba; Glenwood, LLC, Glenwood, NJ), oral prednisone, and topical clobetasol was started. After several months of follow-up, the patient had obtained only moderate improvement of the clinical findings. PMID- 15891260 TI - Case study: delicate skin cutting: management beyond the skin and implications of superficial habitual self-mutilation. AB - A 16-year-old girl accompanied by her mother was noted to have several thin, well healed brown lines on the distal third of her left flexural forearm (Figure). She admits to having recently made these marks by skin cutting. She feels depressed and at times feels life "is not worth living." She denies alcohol or substance abuse or true suicidal ideation, plan, or intent. Her mother describes her as previously well adjusted and a good student who has become erratic and moody in the past 6 months, with worsening school performance. Her father had a history of depression and alcoholism and died 3 years ago. She is undergoing psychotherapy and pharmacotherapy with sertraline with good results. She has not cut her skin in 3 months. Her mood and behavior have improved. PMID- 15891261 TI - The new drug evaluation process: risk benefit ratio. PMID- 15891262 TI - Protein binding and its potential for eliciting minimal systemic side effects with a novel inhaled corticosteroid, ciclesonide. AB - Freely circulating, protein unbound, active inhaled corticosteroid (ICS) can cause systemic adverse effects. Desisobutyryl-ciclesonide (des-CIC) is the active metabolite of ciclesonide, an effective, novel ICS for persistent asthma. This study examines the free fraction of ciclesonide and des-CIC and determines whether the presence of other agents or disease states affects protein binding. Protein binding of des-CIC (0.5, 5.0, 25, 100, and 500 ng/mL) was determined, using both equilibrium dialysis and ultrafiltration, in plasma from humans (healthy and either renally or hepatically impaired) and several animal species and in the presence of either salicylates or warfarin. Dialyzed samples were analyzed by liquid chromatography with tandem mass spectroscopy to determine both free and bound concentrations of des-CIC. After ultrafiltration, spiked plasma plus H-des-CIC was separated into free and bound fractions by centrifugation and quantified by scintillation counting. Additionally, in another study, protein binding of ciclesonide was determined by equilibrium dialysis. For equilibrium dialysis, the mean percentages of des-CIC (0.5-500 ng/mL) plasma protein binding across species were high, approximately 99%, and no apparent saturation of protein binding was observed. Results were similar for ultrafiltration analysis. Protein binding of des-CIC did not change in the presence of warfarin or salicylates or in the plasma of renally or hepatically impaired patients. The protein binding of ciclesonide was 99.4% in human serum. The very low fraction of unbound des-CIC in the systemic circulation suggests minimal systemic exposure of unbound des-CIC, thus suggesting a low potential for systemic adverse effects after administration of inhaled ciclesonide. PMID- 15891263 TI - A one-year comparison of cost and outcomes of angioplasty in stent and nonstent patients. AB - Most studies on costs and outcomes of stent use in percutaneous transluminal coronary angioplasty (PTCA) have been limited to less than 1 year of follow-up, in-hospital costs, or regional samples. The objective of this study was to compare restenosis rates and all direct medical care costs during the 1-year period following stent PTCA and nonstent PTCA. A nationwide claims database covering 1.7 million lives was used to identify patients with PTCA during 1995 and no PTCA during the previous year. All costs and rate of restenotic events during the 1-year period following PTCA were examined. Symptomatic restenosis was defined as the occurrence of any of the following: repeat angioplasty, coronary artery bypass surgery (CABG), and myocardial infarction (MI). Stents were placed in 304 of 1367 identified patients. Both the rate of restenotic events (stent, 16.1%; no stent, 20.1%) and ischemic disease-related costs (stent, $9207; no stent, $10,498) were lower for patients with a stent during the follow-up period; however, the difference was not significant. Because of the higher cost of placing a stent during the initial procedure, patients with a stent had a significantly higher cost at the end of the 1-year period (stent, $49,245; stent, $40,683). Multivariate analyses further confirmed this finding. Thus, although restenotic events were lower for patients with stents by a nonsignificant margin, follow-up costs did not offset the higher cost of the initial stent placement. PMID- 15891264 TI - The effect of magnesium sulfate on action potential duration and cardiac arrhythmias. AB - To evaluate the electrophysiologic and antiarrhythmic effects, Mg was infused at 15 mg/h (n = 5) or an equal volume of saline (1.2 mL/h) (n = 5) and electrocardiogram and action potential duration (APD) recorded every 15 minutes. Rats were anesthetized with 70 mg/kg pentobarbital intraperitoneally. Mg increased QT 15 +/- 6% on average compared with 1 +/- 4 for saline P < 0.01. Mg increased QT, 0, 0, 6 +/- 4, 13 +/- 5, 16 +/- 4, 23 +/- 5, 29 +/- 8, and 32 +/- 5% over baseline after a 2 hours infusion (P < 0.01). APD increased by 0, 6 +/- 3, 8 +/- 8, 14 +/- 4, 16 +/- 12, 21 +/- 4, 25 +/- 5% change from baseline (P < 0.05). The mean percentage of increase was 12 +/- 8 for the Mg group and 1 +/- 3 for the saline group (P < 0.05). The JT interval also increased after Mg (P < 0.01). After Mg loading, coronary occlusion of the left anterior descending coronary artery was performed. Ventricular premature contractions (VPCs), ventricular tachycardia (VT), and ventricular fibrillation (VF) were frequent in the saline control group, with 2 dying in VF; with only scattered VPCs and short runs of nonsustained VT in the Mg group. The results of these findings indicate that infusion of MgSO4 can prolong the QRS, QT, and JT intervals in the rat and these changes correlate well with arrhythmia suppression. PMID- 15891265 TI - Pharmacokinetics of ranitidine and nizatidine in very elderly patients. AB - This study was undertaken to examine whether the pharmacokinetic profiles of ranitidine and nizatidine, the H2 antagonists, differed with repeated doses in very elderly patients (>80 years old). Ranitidine (150 mg) or nizatidine (150 mg) was given twice daily in 10 very elderly female patients for 14 days. This study was a randomized, crossover design with a wash-out period of 14 days. Pharmacokinetic profiles were determined after first (on day 1) and 27th (on day 14) doses of each agent. The maximum plasma drug concentration (Cmax) and area under the plasma drug concentration-time curve (AUC) were significantly greater after the 27th than after the first dose in the ranitidine but not in the nizatidine trials. The accumulation of ranitidine in plasma tended to be greater than that of nizatidine. These results suggest that nizatidine is safer for the repeated treatment in very elderly patients. PMID- 15891266 TI - Antibiotics and risk of first-time hospitalization for myocardial infarction: a population-based case-control study. AB - There is increasing evidence that inflammation plays an important role in atherosclerosis. Such inflammation is likely related to the presence of infectious organisms. Hence, we examined whether the use of antibiotic drugs decreases the risk of first-time myocardial infarction (MI). We identified 6737 cases of first-time hospitalization for MI, and 67,364 age- and gender-matched, population-based controls during 1991-2002, using data from the County Hospital Discharge Registry and the Civil Registration System of North Jutland County, Denmark. All prescriptions for antibiotics prior to the hospitalization for MI were identified through a prescription database. Conditional logistic regression was used to estimate odds ratios (OR) associated with antibiotic use, adjusted for potential confounding factors including previous discharge diagnoses of hypertension, chronic bronchitis and emphysema, alcoholism, liver cirrhosis, or diabetes mellitus and prescriptions for anti-hypertensive drugs, antidiabetic drugs, lipid-lowering agents, high-dose aspirin, platelet inhibitors, oral anticoagulants, or hormone replacement therapy. The use of any one type of antibiotic in the 3 years before hospitalization was not associated with a decreased risk of MI; the adjusted ORs with corresponding 95% confidence intervals were 1.07, 1.00-1.14 for penicillins; 1.15, 1.00-1.33 for macrolides; 0.95, 0.65-1.39 for tetracyclines; 1.25, 0.84-1.87 for quinolones; and 0.95, 0.80 1.12 for sulfonamides. A slight increase in the risk of MI was seen with the use of more than one type of antibiotic in the preceding 3 years (OR = 1.17, 95% CI = 1.09-1.27). Our findings do not support the hypothesis that the use of antibiotics is associated with a lower risk of first-time MI. PMID- 15891267 TI - Intravenous nesiritide in acute heart failure. AB - The purpose of this study is to evaluate the effect of nesiritide on length of hospital stay, readmission rates, and charges compared with usual care for congestive heart failure (CHF). Using a structured retrospective chart review, we reviewed the records of 127 patients admitted with decompensated CHF, looking at length of stay on initial admission, readmission rates, total hospital days over 3 months, 3-month mortality, pharmacy and hospital charges for the initial admission, and total pharmacy and hospital charges over 3 months. Nesiritide had no effect on initial length of stay, readmission rates, or 3-month mortality. Patients with an ejection fraction >30% who received nesiritide spent more days in the hospital over 3 months than those who received usual care, although there were no differences between the groups in patients with an ejection fraction of < or =30%. Pharmacy and hospital charges for both the initial admission and over 3 months were significantly higher for patients who received nesiritide than for those who received usual care. Patients who received nesiritide incurred significantly higher charges for medical care, but nesiritide did not affect length of stay, readmission rates, or mortality. PMID- 15891268 TI - Effects of acute organophosphate poisoning on thyroid hormones in rats. AB - The aim of this study was to investigate the effects of organophosphate poisoning on thyroid hormones. In this study, male Wistar albino rats weighing 200-225 g were used. The rats were divided into 4 groups. Group 1 (n = 10) was administered 30 mg/kg lethal dose of methamidophos, whereas group 2 (n = 7) was treated with physiologic NaCl (SP). Group 3 (n = 10) was treated with 30 mg/kg of methamidophos. When cholinergic symptoms developed among the rats in group 3, they were treated with 40 mg/kg pralidoxime intraperitoneally (IP) and administered atropine IP until the cholinergic symptoms disappeared. Group 4 (n = 7) was treated with SP. After the cholinergic symptoms appeared among the rats in group 1, intracardiac blood samples were taken. In group 3, blood samples were taken after the cholinergic symptoms had disappeared. Then triiodothyronine (T3), thyroxine (T4), thyroid stimulating hormone (TSH), and plasma choline esterase (PCE) levels were studied by RIA. The Kruskal-Wallis test and Mann-Whitney U test were used for comparison between groups. Bonferroni correction was applied when multiple comparisons were made. T3, T4, and TSH levels decreased in group 1 compared with group 2 (P < 0.01). When the results between groups 3 and 4 were compared, it was found that the T3 and T4 levels in group 3 decreased while the decreases in T3 levels were statistically significant (P < 0.01). When comparing the results of groups 1 and 3, the T4 level was lower in group 1 and the T3 level was higher in group 3 (P < 0.01). The TSH level increased in group 3 after treatment (P < 0.01). Thyroid hormones were affected after acute organophosphate poisoning. Hypothyroidism and sick euthyroid syndrome was observed during poisoning and after treatment. In serious poisoning, there may be a poor prognosis, but more extensive studies will illuminate the issue in depth. PMID- 15891269 TI - Trastuzumab: targeted therapy for the management of HER-2/neu-overexpressing metastatic breast cancer. AB - Breast cancers are a biologically heterogeneous group of mammary tumors with distinct natural histories and varied responses to established therapies. They have long been divided into those that are hormone sensitive [as defined by expression of the estrogen receptor alpha (ERalpha) and/or the progesterone receptor (PR)] and those that are not. Notably, only those breast cancers that express ERalpha and/or PR typically respond to hormonal therapy with tamoxifen, aromatase inhibitors, or the newer agent fulvestrant. More recently, the transmembrane tyrosine kinase receptor HER-2/neu was identified as an oncogene overexpressed by about 30% of breast cancers. These HER-2/neu-overexpressing breast cancers define a subset of breast tumors that are characteristically more aggressive, and women who develop them have a shorter survival. Trastuzumab (Herceptin), a humanized monoclonal antibody specific for HER-2/neu, has revolutionized the management of metastatic HER-2/neu-overexpressing breast cancers. As a single agent, it produces response rates similar to those of many single-agent chemotherapeutic agents active in metastatic breast cancer and has limited toxicity. Combining trastuzumab with chemotherapy can result in synergistic antitumor activity. The clear efficacy of trastuzumab against HER 2/neu-overexpressing metastatic breast cancer has led to a keen interest in testing its role in the management of early breast cancer, and multiple large clinical trials are currently in progress. This review summarizes the available clinical data on the use of trastuzumab in HER-neu-overexpressing breast cancer and briefly highlights emerging opportunities for innovative, trastuzumab-based breast cancer therapies. PMID- 15891270 TI - Important role of proinflammatory cytokines/other endogenous substances in drug induced hepatotoxicity: depression of drug metabolism during infections/inflammation states, and genetic polymorphisms of drug-metabolizing enzymes/cytokines may markedly contribute to this pathology. AB - Analysis of literature data on drug-induced hepatotoxicity reveals that often upper respiratory febrile illnesses and/or inflammation states precede liver injury/diseases related to administration of drugs or hepatotoxicity associated with administration of therapeutic doses of acetaminophen in some genetically predisposed subjects. The goals of this paper are to review the potential role of alterations in the balance between TH1 cells producing cytokines associated with a cell-mediated response and TH2 cells associated with an antibody response, as well as other endogenous substances, eg, growth factors, leading to a shift in immune response to one that may participate in the liver cells injury during administration of certain drugs, especially in subjects with genetic polymorphisms in drug-metabolizing enzymes. The papers cited in this review were selected to illustrate specific issue related to how profuse and dysregulated production of cytokines, growth factors, and/or other endogenous substances during viral/bacterial infections and inflammation states play a role in the development of drug-induced liver injury. Several cases of liver injury related to administration of drugs appear to be initiated or intensified by upper respiratory febrile illnesses and/or inflammation states, which stimulate sometimes dysregulated production of interferon gamma and/or other proinflammatory cytokines/growth factors. This, in turn, results in down regulation of various induced and constitutive isoforms of cytochromes P-450, and other enzymes involved in the metabolism of several exogenous (eg, drugs) and endogenous lipophilic (eg, steroids) substances, thus having an important impact on the alterations in bioactivation and detoxication processes in the body and on the balance between production, utilization, and elimination of endogenous bioproducts of these reactions. Activation of systemic host defense mechanisms results in down-regulation of various enzymes involved in drug metabolism and elimination, as well as in production, utilization, and excretion of many endogenous substances that have beneficial effects on vital processes in the body. It seems that treatment of acute and chronic infections and/or inflammations with, for example, antibacterials not metabolized in the liver, and use of medications that decrease proinflammatory cytokine levels (eg, pentoxifylline, a TNF-alpha synthesis inhibitor, directed against TNF-alpha induced priming of human neutrophils, immunotherapy with IL-4, IL-1 receptor antagonists or factors inducing IL-1ra, dietary supplementation with long-chain n 3 fatty acids, and other antioxidant agents) may perhaps, in some cases, be helpful in the prevention and management of drug-induced hepatotoxicity. Drug mediated injuries may eventually be prevented by screening methods that can identify genetic polymorphism of drug-metabolizing enzymes and gene polymorphisms or RNA-expression profiles of some proinflammatory cytokines before a patient uses a drug. PMID- 15891271 TI - Management of myasthenia gravis. AB - Myasthenia gravis (MS) is an immune-mediated disorder characterized by fluctuating weakness and fatigue of voluntary muscles. The muscular disorder is generalized in 85% and confined to extraocular muscles in 15% of patients. Pathophysiology of MG involves generation of antiacetylcholine receptor antibodies, which leads to a reduction of the number of acetylcholine receptors at the muscular motor endplate. This in turn results in fewer acetylcholine receptors available for stimulation, lower amplitude stimulations, less muscle fiber activation, and the eventual development of weakness in the affected muscles. The diagnostic workup for MS consists of administration of anticholinesterase agents (Tensilon test), repetitive nerve stimulation, Ach-R antibody assay, and single-fiber electromyography. Management of patients with MG includes cholinesterase inhibitors, corticosteroids, thymectomy, immunosuppressants, plasma exchange, and IVIg. PMID- 15891272 TI - Importance of early diagnosis and therapy of acute meningococcal myocarditis: a case report with review of literature. AB - Acute meningococcemia is a clinical syndrome resulting from Neisseria meningitides infection. The clinical presentation is varied, but when associated with myocardial dysfunction, it carries a poor prognosis. We report a case of meningococcal myocarditis causing severe left ventricular systolic dysfunction that improved over a 4-day period after appropriate antimicrobial treatment following prompt diagnosis. The case highlights the importance of prompt recognition and treatment of this disorder. PMID- 15891273 TI - Accidental clonidine patch ingestion in a child. AB - A case of a pediatric ingestion of a clonidine patch is described. Symptoms included lethargy, bradycardia, and miosis. Treatment included naloxone, charcoal administration, and whole bowel irrigation. Gastrointestinal absorption of active drug can result in significant toxicity and treatment should focus on aggressive decontamination techniques. PMID- 15891274 TI - Rare skin disorder complicating doxorubicin therapy: miliaria crystallina. AB - Miliaria is a self-resolving disorder of the eccrine sweat glands caused by blockage of the sweat ducts resulting in the leakage of eccrine sweat into epidermis or dermis. In addition to known predisposing factors such as heat, humidity, and fever, a few drugs (eg, isotretinoin and bethanechol) can also precipitate miliaria. We present here a case of miliaria crystallina that developed in a 65-year-old patient on the fifth day after administration of doxorubicin for multiple myeloma in the absence of known predisposing factors. Miliaria crystallina resolved in 3 days without any complications. To our knowledge, this is the first reported case of doxorubicin-induced miliaria crystallina. PMID- 15891275 TI - Use of reperfusion therapy and cardiovascular drugs in the treatment of 146 men and 54 women with ST-segment elevation myocardial infarction at a University Medical Center. AB - We investigated the treatment of 146 men, mean age 62 years, and 54 women, mean age 69 years, with acute ST-segment elevation myocardial infarction (STEMI) in a university medical center. Coronary revascularization or thrombolytic therapy was given to 143 men (98%) and 52 women (96%) [P = not significant (NS)]. Antiplatelet therapy and antithrombotic therapy were given to 146 men (100%) and 54 women (100%) (P = NS). Beta-blockers were given to 133 men (91%) and 45 women (83%) (P = NS). Angiotensin-converting enzyme inhibitors or angiotensin receptor blockers were given to 122 men (84%) and 42 women (78%) (P = NS). Statins were given to 128 men (88%) and 43 women (80%) (P = NS). Nitrates were given to 94 men (64%) and 36 women (67%) (P = NS). Diuretics were given to 97 men (66%) and 37 women (69%) (P = NS). Calcium channel blockers were given to 26 men (18%) and 12 women (22%) (P = NS). There was no significant difference in the treatment of men versus women with acute STEMI. PMID- 15891289 TI - Differentiated thyroid carcinoma in children and young adults: evaluation of response to treatment. AB - AIM: The aim of this study was to evaluate response to treatment of children and young adults with differentiated thyroid carcinoma (DTC). PATIENTS AND METHODS: Thirty-one children and young adults (27 female, 4 male, <25 years) with DTC were treated with radioiodine between 1987 and 2003. All patients had previously undergone total (or near-total) thyroidectomy with lymph node dissection (if enlarged lymph nodes were present). Initial radioiodine therapy was given 4 to 6 weeks after surgery. Repeated doses were given 4 to 6 weeks after l-thyroxine withdrawal. Effect of therapy was evaluated by radioiodine whole-body scans and serum thyroglobulin levels. RESULTS: Age range at diagnosis was 12 to 25 years (median, 21 years). Follow-up duration range was 16 to 150 months (mean, 60 months). Histologic classification was papillary in 28 (90%) patients (follicular variant in 7) and follicular in 3 (10%). Multifocal carcinoma was found in 9 (29%) patients and neck lymph node metastases in 19 (61%) patients. Fifteen (48%) patients had pulmonary or mediastinal metastases at the time of diagnosis, and 1 developed bone metastases. Lung metastases demonstrated a pattern of diffuse radioiodine uptake in 7 patients and focal uptake in 8. Total radioiodine dose range was 80 to 1086 mCi given in 1 to 7 treatments. Fourteen (45%) patients were disease-free, including 1 patient with pulmonary metastases who achieved complete remission. Three (10%) patients had persistent metastases in neck lymph nodes. Nine (29%) patients with pulmonary metastases showed good response, 4 (13%) had stable persistent disease, and 1 had progression of the disease and died after 11.5 years of follow up. All surviving patients (97%) are asymptomatic and leading normal lives. CONCLUSIONS: Children and young adults with DTC had a high incidence of multifocal disease, lymph node involvement, and pulmonary metastases. Although complete remission of pulmonary metastases after radioiodine therapy was difficult to achieve, a partial response with reduction of metastatic disease was possible. In general, the patients had a good quality of life with no further disease progression and a low mortality rate. PMID- 15891290 TI - Hemodynamic and metabolic changes in a patient with cerebral venous sinus thrombosis: evaluation using O-15 positron emission tomography. AB - The cerebral hemodynamics and oxygen metabolism before and after the recanalization of the occluded sinus were evaluated using O-15 positron emission tomography (PET) scanning in a 21-year-old woman with superior sagittal sinus thrombosis. During the initial presentation, there was increased regional cerebral blood volume (rCBV) in the left frontal and parietal lobes with moderately increased rCBV in the right parietal lobe. Recanalization of the occluded sinus resulted in a normalization of elevated rCBV in the frontal and parietal lobes and improvements of rCBF and regional cerebral metabolic rate for oxygen in the occipital lobe. Clinical improvement after recombinant tissue plasminogen activator therapy was accompanied by restoration of cerebral hemodynamics and oxygen metabolism, although magnetic resonance venography showed only partial recanalization of the occluded sinus. PET evaluation of the cerebral hemodynamics and oxygen metabolism may be of value in gaining further insight into pathophysiological characteristics of human cerebral venous sinus thrombosis. PMID- 15891291 TI - Technetium-99m diphosphonate imaging of psammocarcinoma of probable ovarian origin: case report and literature review. AB - Technetium-99m methylene diphosphonate (Tc-99m MDP) bone scans have long been used by clinicians to diagnose osseous metastases in patients with cancer. However, in several benign and malignant diseases, notably those characterized by extensive soft tissue calcification, Tc-99m MDP may be taken up by the tumor itself. We present a case of a stage IIIC psammoma-rich low-grade serous carcinoma of the ovary, whose identity and extent of disease were first suggested by Tc-99m MDP scintigraphy. The literature concerning this form of cancer, and the use of Tc-99m MDP bone scans to image soft tissue lesions, are reviewed. PMID- 15891292 TI - The role of nuclear medicine in the evaluation of complex regional pain syndrome type I. AB - Chronic pain resulting from complex regional pain syndrome type I (CRPS I), formerly referred to as the reflex sympathetic dystrophy syndrome (RSDS), is a diagnostic challenge to the clinician. It involves multiple organ systems, namely peripheral as well as central nervous, vascular, soft tissue, and skeletal. It usually develops as a consequence of trauma, without nerve injury. Signs and symptoms vary depending on the time since the initiating event, and there is no confirmatory histopathologic diagnosis. This article summarizes the current consensus on the classification, pathophysiology, and diagnostic approaches, emphasizing the role of scintigraphy in the management of this multisystem disorder. PMID- 15891293 TI - FDG PET for discrimination between tumor extension and blood thrombus as a cause for portal vein thrombosis in hepatocellular carcinoma: important role in exclusion of transplant candidacy. PMID- 15891294 TI - F-18 FDG PET demonstration of thyroglossal duct cyst. PMID- 15891295 TI - In pyrexia resulting from occult tuberculosis Ga-67 citrate is still more sensitive than newer agents such as Tc-99m selusomab. PMID- 15891296 TI - Fracture of a bone-graft donor site in the ulna. PMID- 15891297 TI - F-18 FDG uptake in bilateral pulmonary artery leiomyosarcomata, one mimicking a pulmonary embolus. PMID- 15891298 TI - Pulmonary amyloidosis: detection with PET-CT. PMID- 15891299 TI - Myocardial bridging in a patient with exertional chest pain. PMID- 15891300 TI - Isolated left ventricular noncompaction: a diagnosis to consider with left ventricular wall akinesis. PMID- 15891301 TI - Fixed defect on stress myocardial imaging resulting from previous trauma masquerading as coronary artery disease. PMID- 15891302 TI - Metastatic calcinosis in small cell ovarian carcinoma, hypercalcemic type. PMID- 15891303 TI - Tc-99m MDP scintigraphy in a case of idiopathic calcinosis cutis. PMID- 15891304 TI - Simultaneous visualization of a mandibular brown tumor with a large parathyroid adenoma on Tc-99m MIBI imaging. PMID- 15891305 TI - Bochdalek hernia, incidental finding on sestamibi parathyroid scan. PMID- 15891306 TI - Sentinel node imaging of laryngeal cancer using a portable gamma camera with CdTe semiconductor detectors. PMID- 15891307 TI - Incidental detection of rounded atelectasis during tc-99m MIBI scintimammography. PMID- 15891308 TI - Occult bony metastatic disease demonstrated on labeled leukocyte scintigraphy. PMID- 15891309 TI - Malignant ameloblastoma with thoracic vertebral metastasis: PET/CT and MR imaging. PMID- 15891310 TI - Vaginal melanoma mimicking bladder FDG activity in a patient with chronic renal failure. PMID- 15891311 TI - Metastatic hepatic lesions are detected better by delayed imaging with prolonged emission time. PMID- 15891312 TI - Massive bleeding from jejunal varices in a patient with thalassemia major detected by TC-99m red blood cell scintigraphy. PMID- 15891313 TI - Current readings in nuclear medicine. PMID- 15891314 TI - Our Journal, Critical Care Medicine, in 2005: high impact factor, rapid manuscript review, growing submissions, and widespread distribution. PMID- 15891315 TI - Clinical implementation of the ARDS network protocol is associated with reduced hospital mortality compared with historical controls. AB - OBJECTIVE: To assess the impact of implementing a low tidal volume ventilation strategy on hospital mortality for patients with acute lung injury or acute respiratory distress syndrome. DESIGN: Retrospective, uncontrolled study. SETTING: Adult medical-surgical and trauma intensive care units at a major inner city, university-affiliated hospital. PATIENTS: A total of 292 patients with acute lung injury or acute respiratory distress syndrome. INTERVENTIONS: Between the years 2000 and 2003, 200 prospectively identified patients with acute lung injury/acute respiratory distress syndrome were managed by the ARDS Network low tidal volume protocol. A historical control group of 92 acute respiratory distress syndrome patients managed by routine practice from 1998 to 1999 was used for comparison. MEASUREMENTS AND MAIN RESULTS: Patients managed with the ARDS Network protocol had a lower hospital mortality compared with historical controls (32% vs. 51%, respectively; p = .004). Multivariate logistic regression estimated an odds ratio of 0.32 (95% CI, 0.17-0.59; p = .0003) for mortality risk with use of the ARDS Network protocol. Protocol-managed patients had a lower tidal volume (6.2 +/- 1.1 vs. 9.8 +/- 1.5 mL/kg; p < .0001) and plateau pressure (27.5 +/- 6.4 vs. 33.8 +/- 8.9 cm H2O; p < .0001) than historical controls. CONCLUSION: Adoption of the ARDS Network protocol for routine ventilator management of acute lung injury/acute respiratory distress syndrome patients was associated with a lower mortality compared with recent historical controls. PMID- 15891316 TI - Variation in outcomes in Veterans Affairs intensive care units with a computerized severity measure. AB - OBJECTIVE: To quantify the variability in risk-adjusted mortality and length of stay of Veterans Affairs intensive care units using a computer-based severity of illness measure. DESIGN: Retrospective cohort study. SETTING: A stratified random sample of 34 intensive care units in 17 Veterans Affairs hospitals. PARTICIPANTS: A consecutive sample of 29,377 first intensive care unit admissions from February 1996 through July 1997. INTERVENTIONS: Standardized mortality ratio (observed/expected deaths) and observed minus expected length of stay (OMELOS) with 95% confidence intervals were estimated for each unit using a hierarchical logistic (standardized mortality ratio) or linear (OMELOS) regression model with Markov Chain Monte Carlo simulation. We adjusted for patient characteristics including age, admission diagnosis, comorbid disease, physiology at admission (from laboratory data), and transfer status. MEASUREMENTS AND MAIN RESULTS: Mortality across the intensive care units for the 12,088 surgical and 17,289 medical cases averaged 11% (range, 2-30%). Length of stay in the intensive care units averaged 4.0 days (range, mean unit length of stay 3.0-5.9). Standardized mortality ratio of the intensive care units varied from 0.62 to 1.27; the standardized mortality ratio and 95% confidence interval were <1 for four intensive care units and >1.0 for seven intensive care units. OMELOS of the intensive care units ranged from -0.89 to 1.34 days. In a random slope hierarchical model, variation in standardized mortality ratio among intensive care units was similar across the range of severity, whereas variation in length of stay increased with severity. Standardized mortality ratio was not associated with OMELOS (Pearson's r = .13). CONCLUSIONS: We identified intensive care units whose indicators for mortality and length of stay differ substantially using a conservative statistical approach with a severity adjustment model based on data available in computerized clinical databases. Computerized risk adjustment employing routinely available data may facilitate research on the utility of intensive care unit profiling and analysis of natural experiments to understand process and outcome links and quality efforts. PMID- 15891317 TI - Dexmedetomidine does not improve patient satisfaction when compared with propofol during mechanical ventilation. AB - OBJECTIVE: Dexmedetomidine (DEX) may provide a sedation level that enables sleep and communication, with less amnesia and pain medication requirements, during mechanical ventilation. Our study directly assessed patient-perceived satisfaction with coronary artery bypass graft surgery after administration of DEX or propofol for intensive care unit (ICU) sedation. DESIGN: Prospective, randomized clinical study with subsequent questionnaire administration. SETTING: Tertiary care surgical ICU. PATIENTS: A total of 89 adult, nonemergent, coronary artery bypass graft patients with an expected length of intubation of <24 hrs. INTERVENTIONS: Patients were randomized to either DEX or propofol; drug administration was performed via standardized anesthesia and nursing protocols. MEASUREMENTS: Patients reported perceptions of their ICU experience after mechanical ventilation with a modified numerical-scale Hewitt questionnaire, validated specifically for ICU patients. Patients were questioned regarding awareness, recall, generalized comfort, level of pain, ability to interact with healthcare providers and family, feelings of agitation and anxiety, perceived ease of extubation, ability to sleep or rest, and satisfaction with ICU experience. MAIN RESULTS: Groups were well matched at baseline, with a mean +/- sd age of 63.0 +/- 10.4 yrs and weight of 88.7 +/- 16.7 kg. No difference was observed for length of surgery, length of intubation, or ICU stay (p > .05). DEX patients perceived a shorter length of intubation (p = .044). A deeper sedation level was found in the propofol group (p = .021), with similar morphine and midazolam requirements (p = .317). Patient-rated level of overall awareness as a marker of amnesia did not differ between groups (p = .653). The ability to rest or sleep trended toward significance favoring propofol (p = .051). On evaluation of questionnaire ratings, DEX patients expressed more discomfort (p = .046), pain (p = .096), and sleeping difficulty (p = .036). Similar comfort levels were reported during extubation (p = .179). CONCLUSIONS: Despite theoretical advantages of DEX to improve overall patient satisfaction, the two agents provide similar responses to amnesia and pain control. According to our findings, DEX does not seem to have any advantage compared with propofol for short-term sedation after coronary artery bypass graft surgery. PMID- 15891318 TI - How many infections are caused by patient-to-patient transmission in intensive care units? AB - OBJECTIVE: The proportion of intensive care unit (ICU)-acquired infections that are a consequence of nosocomial cross-transmission between patients in tertiary ICUs is unknown. Such information would be useful for the implementation of appropriate infection control measures. DESIGN: A prospective cohort study during 18 months. SETTING: Five ICUs from two university hospitals. PATIENTS: All patients admitted for >/=48 hrs. MEASUREMENT: ICU-acquired infections were ascertained during daily bedside patient and chart reviews. Episodes of potential cross-transmission were identified by highly discriminating genetic typing of all clinical and surveillance isolates of the ten bacterial species most frequently associated with nosocomial infections in ICUs. Isolation of indistinguishable isolates in two or more patients defined potential transmission episodes. MAIN RESULTS: During 28,498 patient days, 431 ICU-acquired infections and 141 episodes of nosocomial transmissions were identified. A total of 278 infections were caused by the ten species that were genotyped, and 41 of these (14.5%) could be associated with transmissions between patients. CONCLUSION: Infections acquired during treatment in modern tertiary ICUs are common, but a causative role of direct patient-to-patient transmission can only be ascertained for a minority of these infections on the basis of routine microbiological investigations. PMID- 15891319 TI - Severe community-acquired pneumonia as a cause of severe sepsis: data from the PROWESS study. AB - OBJECTIVE: To investigate community-acquired pneumonia (CAP) as a cause of severe sepsis in the PROWESS (Recombinant Human Activated Protein C Worldwide Evaluation in Severe Sepsis) trial and to evaluate the effect of drotrecogin alfa (activated) (DrotAA) in this subgroup. DESIGN: Retrospective analysis of the severe CAP subgroup in the PROWESS trial. SETTING: Tertiary care institutions in 11 countries. INTERVENTIONS: DrotAA (n = 850), 24 microg.kg.hr for 96 hrs, or placebo (n = 840). PARTICIPANTS: The 1,690 patients with severe sepsis enrolled in the PROWESS trial. MEASUREMENTS AND MAIN RESULTS: Patients were classified as having CAP if lung was the primary site of infection and if they were enrolled directly from home (private residence) with /=25, Pneumonia Severity Index score of >/=4, or CURB-65 (confusion, urea, respiratory rate, blood pressure, age) score of >/=3. CONCLUSIONS: CAP associated with a high Pneumonia Severity Index score, bacteremia, or an intense coagulation and inflammatory response requiring intensive care unit care were indicators of a high risk of death from severe sepsis. In patients with severe sepsis resulting from CAP, a readily identifiable disease, DrotAA, improved survival compared with placebo. PMID- 15891321 TI - Performance of first responders in simulated cardiac arrests. AB - OBJECTIVE: Survival of in-hospital cardiac arrests depends more on first responders than on cardiac arrest teams. The objective of this study was to determine the adherence to algorithms of cardiopulmonary resuscitation of first responders in simulated cardiac arrests in intensive care. A second objective was to assess the effect of the early vs. late availability of a physician on the performance of nurse-based teams acting as first responders. DESIGN: Prospective study. SETTING: Patient simulator in a tertiary level intensive care unit. PARTICIPANTS: A total of 20 teams consisting of three registered nurses and one resident each. INTERVENTIONS: A simulated witnessed cardiac arrest due to ventricular fibrillation occurred in the presence of one nurse while the remaining two nurses could be called to help. Depending on the time of the residents' arrival, teams were classified as "early" (median arrival 50 secs after the onset of the arrest) or "late" (median arrival 150 secs after the onset of the arrest). MEASUREMENTS AND MAIN RESULTS: In all teams, the recognition of the arrest and the calling for help occurred in a timely fashion. However, a median of 85 secs (interquartile range [IQ], 130 secs) elapsed until the start of cardiac massage and 100 secs (IQ, 45 secs) to the first defibrillation. Once commenced, cardiac massage and mask ventilation were carried out during 61% (IQ, 33%) and 77% (IQ, 23%) of the possible time only. Delays and interruptions were generally not recalled by the participants. Compared with teams with late arriving residents, teams with early arriving residents administered more countershocks: 4.5 (IQ, 2) vs. 3.5 (IQ, 1.5; p = .026). CONCLUSIONS: First responders in intensive care often failed to build a team structure that ensured timely, effective, monitored, and ongoing team activity. The early availability of a physician increased the number of countershocks administered. Self-reporting is unsuitable to reliably assess the quality of cardiopulmonary resuscitation. PMID- 15891322 TI - Heliox improves hemodynamics in mechanically ventilated patients with chronic obstructive pulmonary disease with systolic pressure variations. AB - OBJECTIVE: To test the hypothesis that, compared with air-oxygen, heliox would improve cardiac performance in mechanically ventilated patients with severe chronic obstructive pulmonary disease and systolic pressure variations >15 mm Hg and to determine clinical variables associated with favorable hemodynamic responses to heliox. DESIGN: A prospective interventional study. SETTING: Medical and respiratory intensive care units at a university-affiliated tertiary medical center. PATIENTS: Twenty-five consecutive mechanically ventilated patients with severe chronic obstructive pulmonary disease and acute respiratory failure who had systolic pressure variations >15 mm Hg. INTERVENTIONS: Respiratory and hemodynamic measurements were taken at the following time with the same ventilator setting: a) baseline; b) after 30 mins with heliox; and c) 30 mins after return to air-oxygen. MEASUREMENTS AND MAIN RESULTS: Heliox ventilation decreased intrinsic positive end-expiratory pressure (air-oxygen vs. heliox [mean +/- sd] 13 +/- 4 cm H2O vs. 5 +/- 2 cm H2O, p < .05), trapped lung volume (air oxygen vs. heliox 362 +/- 67 mL vs. 174 +/- 86 mL, p < .05), and respiratory changes in systolic pressure variations (DeltaPP) (air-oxygen vs. heliox 29 +/- 5% vs. 13 +/- 7%, p < .05). In the ten patients with pulmonary arterial catheters, heliox decreased mean pulmonary arterial pressure, right atrial pressure, and pulmonary arterial occlusion pressure and increased cardiac index. Preheliox DeltaPP correlated with the magnitude of reduction in intrinsic positive end-expiratory pressure during heliox ventilation. Age, preheliox Paco2, and ratio of forced expiratory volume at first second to forced vital capacity correlated inversely, whereas preheliox DeltaPP correlated positively with increases in cardiac index. CONCLUSIONS: Heliox may be a useful adjunct therapy in patients with severe chronic obstructive pulmonary disease during acute respiratory failure who have persistent intrinsic positive end-expiratory pressure-induced hemodynamic changes despite ventilator management. PMID- 15891323 TI - Operational performance of validated physiologic scoring systems for predicting in-hospital mortality among critically ill emergency department patients. AB - OBJECTIVES: New Simplified Acute Physiology Score (SAPS) II, Morbidity Probability Model at admission (MPM0 II), and Logistic Organ Dysfunction System (LODS) have all demonstrated high accuracy for predicting mortality in intensive care unit populations. We tested the prognostic accuracy of these instruments for predicting mortality among a cohort of critically ill emergency department patients. DESIGN: Secondary analysis of a randomized controlled trial. SETTING: Urban, tertiary emergency department, census >100,000. PATIENTS: Nontrauma emergency department patients admitted to an intensive care unit, aged >17 yrs, with initial emergency department vital signs consistent with shock (systolic blood pressure <100 mm Hg or shock index >1.0), and with agreement of two independent observers for at least one sign and symptom of inadequate tissue perfusion. INTERVENTIONS: Emergency department variables needed for calculation of each scoring system were prospectively collected, and published formulas were used to calculate the probability of in-hospital death for each scoring system. The main outcome was actual in-hospital mortality. The area under the receiver operating characteristic curve was used to evaluate the predictive ability of each scoring system. MEASUREMENTS AND MAIN RESULTS: Ninety-one of 202 patients (45%) were included. The mean age was 56 +/- 16 yrs, 42% were female, the mean initial systolic blood pressure was 84 +/- 13 mm Hg, and the average length of stay in the emergency department was 4.2 +/- 2.0 hrs. The in-hospital mortality rate was 21%. The area under the receiver operating characteristic curve for calculated probability of in-hospital mortality for SAPS II was 0.72 (95% confidence interval, 0.57-0.87), for MPM0 II 0.69 (95% confidence interval, 0.54 0.84), and for LODS 0.60 (95% confidence interval, 0.45-0.76). CONCLUSIONS: Using variables available in the emergency department, three previously validated intensive care unit scoring systems demonstrated moderate accuracy for predicting in-hospital mortality. PMID- 15891324 TI - Cerebral output of cytokines in patients with pneumococcal meningitis. AB - OBJECTIVE: Patients with acute bacterial meningitis frequently develop sepsis, the hallmark of which is increased plasma cytokine levels. However, it is unknown whether the brain contributes to the intravascular accumulation of cytokines in meningitis. We measured the cerebral output of cytokines to the blood during severe pneumococcal meningitis accompanied by sepsis. DESIGN: Prospective physiologic study. SETTING: Multidisciplinary intensive care unit. PATIENTS: Seven patients (median age, 59; range, 26-72 years) with severe pneumococcal meningitis, as evidenced by a decreased level of consciousness and the need for mechanical ventilation, and concomitant sepsis; and seven healthy volunteers (age, 24; range, 21-29 years). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The cerebral output, defined as the cerebral blood flow multiplied by the jugular-to-arterial concentration difference, was measured individually for the cytokines tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6. Cerebral blood flow was measured by the Kety-Schmidt method using an infusion of Xe, and the concentration of individual cytokines in arterial and jugular bulb blood was measured by enzyme-linked immunosorbent assay. Compared with controls, patients exhibited elevated plasma levels of all three cytokines, particularly interleukin-6, as well as a marked cerebral output of tumor necrosis factor-alpha and interleukin-6. No cytokine output was found in volunteers. CONCLUSIONS: Patients with pneumococcal meningitis and sepsis exhibit a cerebral output of tumor necrosis factor-alpha and interleukin-6, which may contribute to elevating the plasma levels of these cytokines. PMID- 15891325 TI - Effect of carbon dioxide on systemic oxygenation, oxygen consumption, and blood lactate levels after bidirectional superior cavopulmonary anastomosis. AB - OBJECTIVE: We sought to assess the effects of four different CO2 tensions on systemic oxygenation, oxygen consumption, and arterial blood lactate levels early after bidirectional superior cavopulmonary anastomosis. DESIGN: Prospective study. SETTING: Quaternary pediatric cardiac critical care unit. PATIENTS: Nine children aged 2-23 months (median, 7 months). INTERVENTIONS: All patients were sedated, muscle relaxed, and mechanically ventilated. Baseline Paco2 was adjusted to 35 mm Hg by changing tidal volume. CO2 was added via the inlet port of the ventilator to maintain the Paco2 at 45 and 55 mm Hg. Measurements were repeated after discontinuing additional CO2 gas at a Paco2 of 40 mm Hg. Arterial blood gases and lactate were measured at each level of Paco2. We measured oxygen consumption continuously by respiratory mass spectrometry. MEASUREMENTS AND MAIN RESULTS: Mean (95% confidence interval) Paco2 increased from 35 (34-36) to 45 (44 46) to 55 (54-56) mm Hg (4.7 [4.5-4.9] to 6 [5.7-6.3] to 7.3 [7.2-7.4] kPa), arterial pH decreased from 7.43 (7.39-7.47) to 7.35 (7.31-7.39) to 7.28 (7.24 7.32). Pao2 increased from 36 (32-40) to 44 (40-48) to 50 (45-55) mm Hg (4.8 [4.3 5.3] to 5.9 [5.4-6.4] to 6.7 [6.2-7.2] kPa), and oxygen saturation increased from 72% (67-79%) to 77% (73-81%) to 80% (76-84%). Oxygen consumption decreased significantly, with each increase in Paco2, from 146 (125-167) to 132 (112-152) to 126 (107-145) mL.min.m (p = .0001), and lactate decreased from 1.5 (1-2.0) to 1.2 (0.8-1.6) to 0.8 (0.5-1.1) mmol/L (p < .01). These changes returned toward baseline at a Paco2 of 40 mm Hg. CONCLUSIONS: Moderate hypercapnia with respiratory acidosis improved arterial oxygenation and reduced oxygen consumption and arterial lactate levels, thus improving overall oxygen transport in children after bidirectional superior cavopulmonary anastomosis. PMID- 15891326 TI - Use of an inspiratory impedance threshold device on a facemask and endotracheal tube to reduce intrathoracic pressures during the decompression phase of active compression-decompression cardiopulmonary resuscitation. AB - INTRODUCTION: Use of an inspiratory impedance threshold device (ITD) significantly increases coronary perfusion pressures and survival in patients ventilated with an endotracheal tube (ETT) during active compression decompression cardiopulmonary resuscitation. We tested the hypothesis that the ITD could lower intratracheal pressures when attached to either a facemask or ETT. METHODS: An active and sham ITD were randomly applied first to a facemask and then to an ETT during active compression-decompression cardiopulmonary resuscitation in 13 out-of-hospital cardiac arrest patients in a randomized, double-blinded, prospective clinical trial. The compression-to-bag-valve ventilation ratio was 15:2. Airway pressures (surrogate for intrathoracic pressure) were measured with a pressure transducer. A sham and an active ITD were used for 1 min each in a randomized order, first on a facemask and then on an ETT. Statistical analyses were made using Friedman's and Wilcoxon's rank-sum tests. RESULTS: For the primary end point, mean +/- sd maximum negative intrathoracic pressures (mm Hg) during the decompression phase of cardiopulmonary resuscitation were -1.0 +/- 0.73 mm Hg with a sham vs. -4.6 +/- 3.7 mm Hg with an active ITD on the facemask (p = .003) and -1.3 +/- 1.3 mm Hg with a sham ITD vs. 7.3 +/- 4.5 mm Hg with an active ITD on an ETT (p = .0009). Decompression phase airway pressures with the facemask and ETT were not statistically different. CONCLUSIONS: Use of an active ITD attached to a facemask or an ETT resulted in a significantly lower negative intratracheal pressure during the decompression phase of active compression-decompression cardiopulmonary resuscitation when compared with controls. Airway pressures with an ITD on either a facemask or ETT were similar. The ITD-facemask combination was practical and enables rapid deployment of this life-saving technology. PMID- 15891327 TI - Time required for equilibration of arterial oxygen pressure after setting optimal positive end-expiratory pressure in acute respiratory distress syndrome. AB - OBJECTIVE: To evaluate the time course of Pao2 change following the setting of optimal positive end-expiratory pressure (PEEP) in patients with acute respiratory distress syndrome (ARDS). DESIGN: Prospective clinical study. SETTING: Multidisciplinary intensive care unit of a university hospital. PATIENTS: Twenty-five consecutive patients with ARDS. INTERVENTIONS: ARDS was diagnosed during pressure-regulated volume control ventilation with tidal volume of 7 mL/kg actual body weight, respiratory rate of 12 breaths/min, inspiratory/expiratory ratio of 1:2, Fio2 of 1, and PEEP of 5 cm H2O. A critical care attending physician obtained pressure volume curves and determined the lower inflection point. Following a rest period of 30 mins with initial ventilation variables, PEEP was set at 2 cm H2O above the lower inflection point, and serial blood samples were collected during 1-hr ventilation with optimal PEEP. Arterial blood gas analyses were performed at 1, 3, 5, 7, 9, 11, 15, 20, 30, 45, and 60 mins. MEASUREMENTS AND MAIN RESULTS: Twenty-five patients were found eligible for the study. Three patients were excluded due to deterioration of oxygen saturation and hemodynamic instability following the initiation of optimal PEEP. Eight cases (36%) were considered to be of pulmonary origin and 14 cases (64%) of extrapulmonary origin. Optimal PEEP levels were 14 +/- 3 cm H2O and 14 +/- 4 cm H2O in pulmonary and extrapulmonary ARDS, respectively. Pao2 demonstrated a 130 +/- 101% increase at the end of 1-hr period in total study population. This improvement did not differ significantly between pulmonary and extrapulmonary forms of ARDS (135 +/- 118% vs. 127 +/- 95%, p = .8). Mean 90% oxygenation time was found to be 20 +/- 19 mins. In the subset of patients with ARDS of pulmonary origin, 90% oxygenation time was 25 +/- 26 mins, whereas it was 17 +/- 15 mins in patients with ARDS of extrapulmonary origin (p = .8). CONCLUSIONS: Our data showed that 20 mins would be adequate for obtaining a blood gas sample in ARDS patients with pulmonary and extrapulmonary origin after application of optimal PEEP 2 cm H2O above the lower inflection point. PMID- 15891328 TI - NH2 terminal pro-brain natriuretic peptide plasma level as an early marker of prognosis and cardiac dysfunction in septic shock patients. AB - OBJECTIVE: To investigate N-terminal pro-brain natriuretic peptide (NT-proBNP) level as a prognostic factor and a marker of myocardial dysfunction in patients with septic shock. DESIGN: Prospective observational study. SETTING: Intensive care unit. SUBJECTS: A total of 39 patients diagnosed with septic shock and requiring mechanical ventilation. MEASUREMENTS AND MAIN RESULTS: Demographic, hemodynamic, respiratory, and biological data (notably NT-proBNP, lactate, and cardiac troponin I) were collected at inclusion and every 12 hrs. The independent factors for death were higher Sequential Organ Failure Assessment score in the 24 hr period after inclusion (odds ratio, 4.7; 95% confidence interval, 1.15-19.3) and the highest NT-proBNP level in the 24-hr period after inclusion (odds ratio, 1.12 per 1000 pg/mL increase; 95% confidence interval, 1.05-1.26). An NT-proBNP of >13,600 pg/mL predicted intensive care unit mortality with an accuracy of 77%. Area under the receiver operating characteristic curve was 0.8 (p = .002; 95% confidence interval, 0.66-0.93). NT-proBNP levels were over the accepted normal range in all patients. Values were highest between 24 and 36 hrs after onset of septic shock and were significantly higher in nonsurvivors at each time between inclusion and day 7. The lowest left ventricular stroke work index of the first 24-hr period after inclusion was the only factor that independently influenced higher NT-proBNP levels at the same time (odds ratio, 0.91; 95% confidence interval, 0.84-0.98). CONCLUSION: NT-proBNP seems to be an early factor of prognosis and myocardial dysfunction in patients with septic shock. PMID- 15891329 TI - Lack of alteration of endogenous nitric oxide pathway during prolonged nitric oxide inhalation in intensive care unit patients. AB - OBJECTIVE: To compare hemodynamic and gasometric variables and the plasma concentrations of nitric oxide metabolites (cyclic guanosine monophosphate and nitrate and nitrite), endothelin-1, and renin-angiotensin metabolites before and after the start of nitric oxide inhalation, after prolonged nitric oxide inhalation, and before and after nitric oxide withdrawal. DESIGN: Prospective study. SETTING: Surgical intensive care unit, university hospital. SUBJECTS: Patients with acute lung injury and right ventricular failure. INTERVENTIONS: Nitric oxide inhalation (10-12 ppm) during a median of 2.9 days (12 hrs to 6.5 days). MEASUREMENTS AND MAIN RESULTS: The pulmonary vasodilator effects of inhaled nitric oxide improved arterial oxygenation in patients with acute lung injury (p < .05) and reduced right atrial pressure in patients with right ventricular dysfunction (p < .01). These beneficial effects lasted the whole period of prolonged inhaled nitric oxide therapy up to 6.5 days. However, when inhaled nitric oxide was withdrawn, pulmonary vasodilator effects rapidly disappeared, and Pao2/Fio2 ratio markedly deteriorated in all studied patients to return to pre-inhaled nitric oxide levels. Changes in plasma cyclic guanosine monophosphate and nitrate and nitrite paralleled those of pulmonary vasodilatory effects. An immediate increase in plasma cyclic guanosine monophosphate with a slightly delayed increase in plasma nitrate and nitrite was observed at inhaled nitric oxide start with no attenuation during the prolonged inhaled nitric oxide therapy. A marked decrease toward pre-inhaled nitric oxide levels was seen within hours of inhaled nitric oxide withdrawal. In addition, no alteration of plasma endothelin-1 or renin-angiotensin mediators was observed during or after inhaled nitric oxide therapy. CONCLUSIONS: Our study showed a lack of attenuation in the beneficial effects of inhaled nitric oxide and a lack of alteration of endogenous nitric oxide, endothelin-1, and renin-angiotensin pathways during prolonged nitric oxide inhalation. PMID- 15891330 TI - Percutaneous translaryngeal versus surgical tracheostomy: A randomized trial with 1-yr double-blind follow-up. AB - OBJECTIVE: To compare the outcomes and the short- and long-term complications of percutaneous translaryngeal tracheostomy (TLT) and surgical tracheostomy (ST). DESIGN: Prospective, randomized clinical trial with 1-yr double-blind follow-up. SETTING: A general intensive care unit of a university hospital. PATIENTS: A total of 139 consecutive critically ill patients who required a tracheostomy between February 2001 and June 2002 were randomly assigned to receive either ST or TLT. RESULTS: TLTs were performed more rapidly than STs (17 +/- 10 mins vs. 22 +/- 6 mins, p = .003). Early complications were rare in both groups. Major postoperative bleeding was less frequent with TLT (0 [0%] vs. 6 [8%], p = .03). Only one case of bleeding (in the ST group) required blood transfusion. Immediately after tracheostomy, six TLT patients (9%) and six patients (8%) in the ST group (p = .56) developed culture-confirmed bacteremia with microbes previously isolated from the pharynx or trachea. Group rates for stomal infections and pneumonia after tracheostomy were similar. At 1-yr follow-up, the overall survival rate was 27%, and 14 patients (45% of survivors) still had open tracheostomies. Both groups rated their quality of life as moderately to severely compromised, and the deterioration was strictly related to the presence of tracheostomy. One TLT and two ST survivors (p = .53) had clinical signs of tracheal stenosis, and bronchoscopy revealed narrowing of >50%. CONCLUSIONS: Compared with ST, the main advantages of TLT are that it is more rapid and associated with less postoperative bleeding. Infectious complications, particularly postoperative bacteremia, and long-term effects (physical and emotional) are similar with the two procedures. PMID- 15891331 TI - Tumor necrosis factor-alpha-induced caspase activation mediates endotoxin-related cardiac dysfunction. AB - OBJECTIVE: Sepsis-induced cardiac dysfunction is a serious clinical syndrome characterized by hypotension, decreased systemic vascular resistance, and elevated cardiac index. Although cytokines such as tumor necrosis factor (TNF) alpha have been shown to play a significant role early in this response, the downstream effects of TNF-alpha signaling on cardiac function, specifically its relationship to apoptosis, have not been fully elucidated. DESIGN: Previous studies from our laboratory have identified endotoxin-induced apoptosis in cardiac cells in vitro. To further determine the role of lipopolysaccharide induced apoptosis in vivo, mice were injected intraperitoneally with lipopolysaccharide (4 mg/kg), and cardiac apoptosis was detected and inhibited using a broad-spectrum caspase inhibitor. SETTING: University research laboratory. SUBJECTS: Adult male wild-type (B6:129PF1/J) and TNF receptor 1/receptor 2 (TNFR-1/2) knockout mice (B6;129S-Tnfrsf1aTnfrsf1b). INTERVENTIONS: We sought to determine the dependence of cardiac apoptosis on TNF-alpha signaling and determine the physiologic role of caspase activation on lipopolysaccharide induced cardiac dysfunction. MEASUREMENTS AND MAIN RESULTS: Cardiac apoptosis was determined at baseline and at 2, 4, 8, and 24 hrs by detection of capase-3 and -8 activity, cytoplasmic levels of Bax/Bcl-2, cleaved caspase-3 immunohistochemistry, and terminal deoxynucleotidyl transferase UTP nick-end labeling (TUNEL) staining of histologic sections in wild-type and TNFR-1/2 knockout mice. To determine the role of caspase activation in lipopolysaccharide induced cardiac dysfunction, a broad-spectrum caspase inhibitor Z-Val-Ala-Asp (ome)-FMK (sad) was given, and cardiac function was determined in isolated beating hearts (Langendorff preparation). Our experiments determined that caspase 3-dependent apoptosis was active in cardiac tissue by 2 hrs and that this activation was completely mediated by TNFR-1/2. The Bax/Bcl-2 ratios supported the finding and time course of apoptosis, whereas TUNEL staining of cardiac tissue sections identified sporadic apoptotic ventricular cells. The administration of zVAD significantly inhibited myocardial caspase-3 activity and preserved cardiac physiologic function (Langendorff preparation). CONCLUSIONS: Endotoxin induces a TNF-alpha-dependent apoptotic cascade in the myocardium, which contributes to the development of cardiac dysfunction. PMID- 15891332 TI - The impact of mechanical ventilation on the moxifloxacin treatment of experimental pneumonia caused by Streptococcus pneumoniae. AB - OBJECTIVE: Streptococcus pneumoniae is a leading cause of community-acquired pneumonia and is responsible for early-onset ventilator-associated pneumonia as well. In intensive care units, community-acquired pneumonia is still associated with a mortality rate of up to 30%, especially when mechanical ventilation is required. Our objective was to study to what extent MV could influence the efficacy of moxifloxacin in a rabbit model of pneumonia. DESIGN: Prospective experimental study. SETTING: University hospital laboratory. SUBJECTS: Male New Zealand White rabbits (n = 75). INTERVENTIONS: S. pneumoniae (16089 strain; minimal inhibitory concentration for moxifloxacin = 0.125 mg/L) was instilled intrabronchially. Four hours later, a human-like moxifloxacin treatment was initiated in spontaneously breathing (SB) and mechanically ventilated (MV) animals. Untreated rabbits were used as controls. Survivors were killed 48 hrs later. Pneumonia was assessed and moxifloxacin pharmacokinetics were analyzed. MEASUREMENTS AND MAIN RESULTS: Moxifloxacin treatment was associated with an improvement in survival in the SB animals (13 of 13 [100%] vs. eight of 37 [21.6%] controls). The survival rate was less influenced by treatment in MV rabbits (seven of 15 [46.1%] vs. one of eight [12.5%] controls). The lung bacterial burden was greater in MV compared with SB rabbits (5.1 +/- 2.4 vs. 1.6 +/- 1.4 log10 colony-forming units/g, respectively). Nearly all the untreated animals presented bacteremia as reflected by a positive spleen culture. No bacteremia was found in SB animals treated with moxifloxacin. In contrast, three of 13 (23.1%) moxifloxacin-treated and MV animals had positive spleen cultures. The apparent volume of distribution of moxifloxacin was lower in MV compared with SB rabbits. CONCLUSIONS: In our model of moxifloxacin-treated S. pneumoniae pneumonia, mechanical ventilation was associated with a higher mortality rate and seemed to promote bacterial growth as well as systemic spread of the infection. In addition, the volume of distribution of moxifloxacin was reduced in the presence of mechanical ventilation. Although the roles of factors such as anesthesia, paralysis, and endotracheal tube insertion could not be established, these results suggest that mechanical ventilation may impair host lung defense, rendering antibiotic therapy less effective. PMID- 15891333 TI - Toll-like receptor 4 deficiency and acute pancreatitis act similarly in reducing host defense during murine Escherichia coli peritonitis. AB - OBJECTIVE: Acute pancreatitis is frequently complicated by Gram-negative sepsis. Mammalian cells recognize lipopolysaccharide from Gram-negative bacteria via Toll like receptor (TLR) 4. The objective of this study was to determine the role of TLR4 in the defense against Gram-negative sepsis in previously healthy mice and in animals with preexisting pancreatitis. DESIGN: A controlled, in vivo laboratory study. SETTING: Research laboratory of a health sciences university. SUBJECTS: Female C3H/HeJ (nonfunctional TLR4 mutant) and C3H/HeN (wild-type) mice. INTERVENTIONS: Abdominal sepsis was induced by the intraperitoneal injection of Escherichia coli. Pancreatitis was induced by 12 hourly intraperitoneal injections of cerulein. MEASUREMENTS AND MAIN RESULTS: The following experiments were performed. First, healthy TLR4 mutant mice demonstrated an enhanced bacterial load and dissemination of the infection relative to wild-type mice after intraperitoneal injection with E. coli, associated with a reduced early release of proinflammatory cytokines and an attenuated influx of neutrophils into the peritoneal fluid. Second, wild-type mice in which acute pancreatitis was induced by repeated cerulein injections showed an increased bacterial load and dissemination of E. coli relative to wild type mice without pancreatitis, which was accompanied by a blunted proinflammatory cytokine response by peritoneal macrophages ex vivo and a diminished early cytokine and neutrophil response in vivo. Third, whereas the severity of cerulein-induced pancreatitis was similar in TLR4 mutant and wild type mice, the important contribution of TLR4 to an effective host defense against E. coli sepsis observed in previously healthy mice was no longer present in mice with preexisting pancreatitis. CONCLUSIONS: These data suggest that TLR4 deficiency and acute pancreatitis act similarly in reducing host defense against E. coli peritonitis and that the role of TLR4 in severe Gram-negative infection depends, at least in part, on the presence of preexisting critical illness. PMID- 15891334 TI - Ketamine reduces nitric oxide biosynthesis in human umbilical vein endothelial cells by down-regulating endothelial nitric oxide synthase expression and intracellular calcium levels. AB - OBJECTIVE: Ketamine, an intravenous anesthetic agent, can modulate vascular tone. Nitric oxide (NO), constitutively produced in endothelial cells, contributes to vasoregulation. In this study, we attempted to evaluate the effects of ketamine on NO biosynthesis in human umbilical vein endothelial cells and its possible mechanism. DESIGN: Controlled laboratory study SETTINGS: Research laboratory in a universal hospital. SUBJECTS: Human umbilical vein endothelial cells prepared from human umbilical cord veins were exposed to 1, 10, 100, and 1000 microM ketamine for 1, 6, and 24 hrs. MEASUREMENTS AND MAIN RESULTS: Exposure to 1, 10, and 100 microM ketamine for 1, 6, and 24 hrs was not cytotoxic to human umbilical vein endothelial cells. However, ketamine at 1000 microM significantly caused cell apoptosis. A therapeutic concentration of ketamine (100 microM) time dependently reduced the levels of nitrite in human umbilical vein endothelial cells. Immunoblot analysis revealed that ketamine time-dependently decreased endothelial NO synthase protein production in human umbilical vein endothelial cells. Results of an assay by reverse-transcription polymerase chain reaction showed that ketamine significantly inhibited levels of endothelial NO synthase messenger RNA. Ketamine time-dependently reduced bradykinin-enhanced intracellular calcium concentrations. Analysis by confocal microscopy further demonstrated the suppressive effects of ketamine on bradykinin-induced calcium mobilization. CONCLUSIONS: A clinically relevant concentration of ketamine can reduce NO biosynthesis. The suppressive mechanisms occur not only by pretranslational inhibition of eNOS expression but also by a posttranslational decrease in endothelial NO synthase activity due to a reduction in intracellular calcium levels. PMID- 15891335 TI - Tetrahydrobiopterin prevents platelet-activating factor-induced intestinal hypoperfusion and necrosis: Role of neuronal nitric oxide synthase. AB - OBJECTIVE: We reported previously that neuronal nitric oxide synthase (nNOS) is the predominant NOS in rat small intestine and is down-regulated by platelet activating factor (PAF). The severity of the bowel injury induced by PAF is inversely related to its suppressing effect on nNOS. Here, we investigated whether intestinal perfusion is regulated by nNOS and whether tetrahydrobiopterin, a co-factor and stabilizer of nNOS, reverses PAF-induced intestinal hypoperfusion and injury. SETTING: Animal laboratory. DESIGN: We first examined nNOS regulation of splanchnic blood flow by measuring the perfusion of the heart, lung, ileum, and kidney in rats after a nNOS inhibitor. We then examined the protective effect of tetrahydrobiopterin on PAF-induced bowel injury, mesenteric hypoperfusion, and systemic inflammation. SUBJECTS: Adult male Sprague-Dawley rats. INTERVENTION: In part 1 of the experiment, rats were given 7 nitroindazole (a specific nNOS inhibitor, 50 mg.kg.day). In part 2 of the experiment, rats were treated with tetrahydrobiopterin (20 mg/kg) 5 mins before and 30 mins after PAF challenge (2.2 microg/kg, intravenously) MEASUREMENTS: Perfusion of the heart, lung, ileum, and kidney was measured at 1 and 4 days after 7-nitroindazole, using fluorescent microspheres. Intestinal injury and inflammation (myeloperoxidase content), blood perfusion, calcium dependent-NOS activity, and systemic inflammation (hypotension and hematocrit increase) were assessed 1 hr after PAF with and without tetrahydrobiopterin treatment. RESULTS: In part 1 of the experiment, 7-nitroindazole induced a long-lasting reduction of blood perfusion and inducible NOS expression selectively in the ileum but not in nonsplanchnic organs such as heart, lungs, and kidneys. In part 2, tetrahydrobiopterin protected against PAF-induced intestinal necrosis, hypoperfusion, neutrophil influx, and NOS suppression. It also reversed hypotension and hemoconcentration. Sepiapterin (2 mg/kg, stable tetrahydrobiopterin precursor) also attenuated PAF-induced intestinal injury. CONCLUSIONS: We conclude that nNOS selectively regulates intestinal perfusion. Tetrahydrobiopterin prevents PAF-induced intestinal injury, probably by stabilizing nNOS and maintaining intestinal perfusion. PMID- 15891336 TI - Effects of tempol, a free radical scavenger, on long-term hyperdynamic porcine bacteremia. AB - OBJECTIVES: Pretreatment with tempol, a membrane-permeable radical scavenger, has been shown to be protective in rodent models of endotoxic and Gram-positive shock. However, neither the pretreatment design nor hypodynamic endotoxic shock in rodents mimics the clinical scenario. Therefore, we investigated the effects of tempol in a posttreatment model of long-term, volume-resuscitated, hyperdynamic porcine bacteremia. DESIGN: Prospective, randomized, controlled experimental study. SETTING: University animal laboratory. SUBJECTS: Sixteen anesthetized, mechanically ventilated, and instrumented pigs. INTERVENTIONS: Sepsis was induced and maintained for 24 hrs with continuous infusion of live Pseudomonas aeruginosa. After 12 hrs of hyperdynamic sepsis, animals were randomized to receive either vehicle (control, n = 8) or continuous infusion of tempol (n = 8, 30 mg/kg/hr). MEASUREMENTS AND MAIN RESULTS: Systemic and hepatosplanchnic hemodynamics, oxygen exchange, metabolism, ileal mucosal microcirculation, and tonometry as well as oxidative stress and coagulation variables were assessed before and after 12, 18, and 24 hrs of P. aeruginosa infusion. Tempol significantly attenuated reduction in mean arterial pressure. Despite comparable mesenteric macrocirculation, tempol attenuated the otherwise progressive deterioration in ileal mucosal microcirculation and prevented mucosal acidosis. By contrast, treatment with tempol failed to influence the P. aeruginosa-induced derangements of hepatosplanchnic redox state, liver lactate clearance, and regional acidosis but prevented the development of renal dysfunction. In addition, tempol reduced nitrosative stress without significant effect on the gradual increase in plasma 8-isoprostanes. Finally, tempol attenuated sepsis-induced endothelial (von Willebrand factor) and hemostatic dysfunction (thrombin-antithrombin complexes, plasminogen activator inhibitor type 1). CONCLUSIONS: The radical scavenger tempol partially prevented live bacteria from causing key features of hemodynamic and metabolic derangements in porcine hyperdynamic sepsis and beneficially affected surrogate markers of sepsis induced endothelial and coagulation dysfunction. Incomplete reduction of oxidative stress because of dilutional effects and/or missed optimal therapeutic window for antioxidant treatment when used in posttreatment approach may account for the only partial protection by tempol in this model. PMID- 15891337 TI - A free radical scavenger, edaravone (MCI-186), diminishes intestinal neutrophil lipid peroxidation and bacterial translocation in a rat hemorrhagic shock model. AB - OBJECTIVE: To investigate the effects of edaravone, a novel free radical scavenger, on bacterial translocation induced by hemorrhagic shock. DESIGN: Prospective, randomized, unblinded animal study. SETTING: Surgical research laboratories of Shiga University of Medical Science. SUBJECTS: Male specific pathogen-free Sprague-Dawley rats. INTERVENTIONS: The rats were randomly divided into three groups: conventional saline treatment, edaravone treatment, and sham shock induction. The saline and edaravone groups were subjected to hemorrhagic shock (mean arterial pressure of 30 mm Hg, for 30 or 60 mins). Rats were killed 30 or 60 mins after shock induction. Mesenteric lymph nodes were cultured for determination of bacterial translocation. Systemic plasma silkworm larvae plasma test, which can detect peptidoglycan and beta-glucan, and endotoxin tests were performed. Immunohistochemistry for 4-hydroxy-2-nonenal (4-HNE) was used to assess lipid peroxidation after shock. MEASUREMENTS AND MAIN RESULTS: The incidence and magnitude of hemorrhagic-shock-induced bacterial translocation to mesenteric lymph nodes were reduced by edaravone. Hemorrhagic-shock-induced increase of plasma silkworm larvae plasma test was also reduced by edaravone. Immunohistochemistry for 4-HNE showed many 4-HNE-positive cells in the lamina propria of the ileum 60 mins after hemorrhagic shock. Double immunohistochemistry revealed that many of these 4-HNE-positive cells were also myeloperoxidase positive. Moreover, the percentage of double-labeled cells with 4-HNE and myeloperoxidase in myeloperoxidase-positive cells was significantly lower in the edaravone group than in the saline group. CONCLUSIONS: The present findings suggest that lipid peroxidation of intestinal neutrophils is involved in bacterial translocation during hemorrhagic shock and that edaravone is potentially useful in diminishing bacterial translocation after hemorrhagic shock. PMID- 15891338 TI - Recruitment maneuvers attenuate repeated derecruitment-associated lung injury. AB - OBJECTIVE: Repeated derecruitments of previously recruited lungs can exacerbate lung injuries during mechanical ventilation. The aim of this study was to assess lung injury associated with repeated derecruitments and to assess whether this type of injury could be attenuated by recruitment maneuvers. DESIGN: Prospective, randomized, experimental animal study. SETTING: University laboratory. SUBJECTS: New Zealand White rabbits. INTERVENTIONS: Twenty-one rabbits were ventilated in pressure-controlled mode with constant tidal volume (10 mL/kg). After lung injury was induced by repeated saline lavage, positive end-expiratory pressure (PEEP) at a lower inflection point was applied for 3 hrs. The control group (n = 7) received ventilation with the same PEEP for 3 hrs without derecruitments. In the derecruitment group (n = 7), derecruitment was repeatedly induced by intentional disconnection of the ventilatory circuit for 1 min every 10 mins for 3 hrs. In the recruitment maneuver group (n = 7), continuous positive airway pressure of 30 cm H2O was applied for 30 secs after each derecruitment. MEASUREMENTS AND MAIN RESULTS: After PEEP levels were increased to lower the inflection point value, Pao2 increased to >500 mm Hg in all groups. Increased Pao2 persisted at >450 mm Hg in the control and recruitment maneuver groups, whereas progressive declines in arterial oxygen levels were observed in the derecruitment group (median, 381.1 mm Hg [interquartile range, 350.1-466.7 mm Hg] at 2 hrs and 318.2 mm Hg [214.3 414.9 mm Hg] at 3 hrs, p < .05 compared with other groups). Histologically, there was significantly increased hyaline membrane formation in alveolar ducts in the derecruitment group compared with the control group (p = .005). Also, significantly more membranous and respiratory bronchiolar injuries were observed in the derecruitment group compared with the control and recruitment maneuver group (p < .005). CONCLUSIONS: These findings suggest that repeated derecruitments could induce lung injuries during mechanical ventilation, and recruitment maneuvers may attenuate derecruitment-associated lung injuries. PMID- 15891339 TI - Massive brain injury enhances lung damage in an isolated lung model of ventilator induced lung injury. AB - OBJECTIVE: To assess the influence of massive brain injury on pulmonary susceptibility to injury attending subsequent mechanical or ischemia/reperfusion stress. DESIGN: Prospective experimental study. SETTING: Animal research laboratory. SUBJECTS: Twenty-four anesthetized New Zealand White rabbits randomized to control (n = 12) or induced brain injury (n = 12) group. INTERVENTIONS: After randomization, brain injury was induced by inflation of an intracranial balloon-tipped catheter, and animals were ventilated with a tidal volume of 10 mL/kg and zero end-expiratory pressure for 120 mins. Following heart lung block extraction, isolated and perfused lungs were subjected to injurious ventilation with peak airway pressure 30 cm H2O and positive end-expiratory pressure 5 cm H2O for 30 mins. MEASUREMENTS AND MAIN RESULTS: No difference was observed between groups in gas exchange, lung mechanics, or hemodynamics during the 2-hr in vivo period following induction of brain injury. However, after 30 mins of ex vivo injurious mechanical ventilation, lungs from the brain injury group showed greater change in ultrafiltration coefficient, weight gain, and alveolar hemorrhage (all p < .05). CONCLUSIONS: Massive brain injury might increase lung vulnerability to subsequent injurious mechanical or ischemia reperfusion insults, thereby increasing the risk of clinical posttransplant graft failure. PMID- 15891340 TI - Hyperglycemia enhances the cytokine production and oxidative responses to a low but not high dose of endotoxin in rats. AB - OBJECTIVE: The aim of this study was to investigate whether hyperglycemia enhances the systemic inflammatory response and oxidative stress induced by endotoxin. DESIGN: Laboratory investigation. SETTING: University medical school. SUBJECTS: Forty-one male Sprague-Dawley rats. INTERVENTIONS: A hyperglycemic condition was produced in rats by glucose clamp for 3 hrs. Immediately on stopping the glucose infusion, animals received different doses of endotoxin injection (0, 0.2, or 1 mg/kg), and then blood glucose concentration was monitored over the ensuing 2 hrs. At the end of 2 hrs, levels of tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, corticosterone, and alpha-1 acid glycoprotein were determined in serum, and malondialdehyde and total glutathione content were determined in the liver. MEASUREMENTS AND MAIN RESULTS: Over the 2 hr period, blood glucose concentrations returned to normal in initially hyperglycemic rats. However, the levels of cytokines, corticosterone, and alpha-1 acid glycoprotein were significantly higher in these animals compared with nonhyperglycemic controls, demonstrating an extended effect of prior hyperglycemia on markers of systemic inflammation. With low-dose (0.2 mg/kg) but not high-dose (1 mg/kg) endotoxin administration, hyperglycemic animals had significantly higher levels of cytokines compared with controls, indicating that prior hyperglycemia can enhance the systemic inflammatory response to a moderate endotoxin dose, but that the maximum effects of endotoxin on production of inflammatory cytokines are not altered by transient high glucose exposure. CONCLUSIONS: Systemic inflammation persists for a period following hyperglycemia, and this can enhance the systemic inflammatory response to a subsequent moderate stress. PMID- 15891341 TI - Spontaneous breathing affects the spatial ventilation and perfusion distribution during mechanical ventilatory support. AB - OBJECTIVE: In acute respiratory failure, gas exchange improves with spontaneous breathing during airway pressure release ventilation (APRV). The mechanisms for this improvement are not fully clear. We have shown that APRV with spontaneous breathing reopens nonaerated lung tissue in dorsal juxtadiaphragmatic regions. We hypothesized that spontaneous breathing during APRV may redistribute ventilation and perfusion toward these reopened regions. DESIGN: Prospective, randomized, controlled study. SETTING: Animal research laboratory SUBJECTS: Twenty controlled mechanically ventilated pigs. INTERVENTIONS: Lung injury was induced by injection of oleic acid into the central circulation; thereafter, pigs were randomized to APRV with or without spontaneous breathing. To induce spontaneous breathing during APRV with spontaneous breathing, the mechanical respiratory rate was decreased by 50% in this group. MEASUREMENTS AND MAIN RESULTS: We measured respiratory mechanics, hemodynamics, gas exchange including the multiple inert gas elimination technique, and the spatial ventilation and perfusion distribution using single photon emission tomography. At similar minute ventilation and airway pressures, shunt remained stable during APRV with spontaneous breathing, whereas it increased during APRV without spontaneous breathing during the 2-hr study period (p = .006). Single photon emission tomography showed more ventilation (p < .001) and pulmonary blood (p < .025) flow in dorsal, juxtadiaphragmatic lung regions when spontaneous breathing was present. CONCLUSIONS: The beneficial effects of spontaneous breathing on intrapulmonary shunt and oxygenation are explained both by increased ventilation of aerated dependent lung tissue and by opening up nonaerated tissue so that ventilation is distributed to a larger share of the lung. Redistribution of perfusion is possibly secondary to the altered ventilation. The overall effect is a more efficient use of available lung tissue for gas exchange. PMID- 15891342 TI - Differential impact of perfluorochemical physical properties on the physiologic, histologic, and inflammatory profile in acute lung injury. AB - OBJECTIVE: To evaluate the differential effects of physical properties of combinational perfluorochemical liquids (PFC) during partial liquid ventilation (PLV) on inflammatory indexes in the injured lung. DESIGN: : Interventional laboratory study. SETTING: Academic medical research laboratory. SUBJECTS: Seventeen saline lavage-injured juvenile rabbits. INTERVENTIONS: Rabbits were anesthetized, ventilated, saline lavage-injured, and randomized into groups: group 1 (conventional mechanical ventilation alone-no PFC), group 2 (PLV: lowest viscosity, highest vapor pressure), group 3 (PLV: mid-viscosity, mid-vapor pressure), group 4 (PLV: highest viscosity, lowest vapor pressure). MEASUREMENTS AND MAIN RESULTS: Arterial blood chemistry and pulmonary mechanics were monitored throughout the protocol. Following 4 hrs, lung tissue was harvested for interleukin-8, myeloperoxidase, and histologic analyses. Oxygenation (Pao2), ventilation (ventilation efficiency index), and respiratory compliance were not significantly different between groups before or following injury. Pao2 increased significantly following treatment in groups 3 and 4. Oxygenation index was significantly lower and respiratory compliance and ventilation efficiency index were significantly higher for group 4 following 4 hrs than all other groups. Total lung tissue interleukin-8 was significantly lower in groups 3 and 4 than groups 1 and 2, and lung myeloperoxidase was significantly lower in all PLV treated groups than CMV alone. Histologic examination showed increased recruitment of the dependent lung in groups 3 and 4, with significantly greater lung expansion index, than groups 1 and 2. CONCLUSIONS: PLV, with a single dose of higher viscosity and lower vapor pressure PFC, resulted in significantly improved gas exchange and lung mechanics with significant reduction in lung inflammation compared with conventional mechanical ventilation alone and PLV with lower viscosity and higher vapor pressure liquid. Since PFC evaporative loss and redistribution are minimized by lower VP and higher viscosity, these data suggest that greater mechanoprotection and cytoprotection of the lung are conferred during PLV with PFC liquids that remain distributed throughout the entire lung for a longer duration. PMID- 15891343 TI - Packed red blood cell transfusion increases local cerebral oxygenation. AB - OBJECTIVE: To determine a) whether packed red blood cell transfusion (RBCT) increases local brain tissue oxygen partial pressure (Pbto2) in a neurocritical care population; and b) what (if any) demographic, clinical, or physiologic variables mediate the assumed change. DESIGN: Prospective observational study. SETTING: A neurosurgical intensive care unit at a university-based level I trauma center and tertiary care hospital. PATIENTS: Thirty-five consecutive volume resuscitated patients with subarachnoid hemorrhage or traumatic brain injury, without cardiac disease, requiring Pbto2 monitoring and receiving RBCT were studied between October 2001 and December 2003. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The following physiologic variables were measured and compared 1 hr before and after RBCT: Pbto2, intracranial pressure, cerebral perfusion pressure, hemoglobin oxygen saturation (Sao2), Fio2, hemoglobin, and hematocrit. An increase in Pbto2 was observed in 26 of the 35 patients (74%). In nine patients, Pbto2 decreased after RBCT. The mean (+/-sd) increase in Pbto2 for all patients was 3.2 +/- 8.8 mm Hg (p = .02), a 15% change from baseline (1 hr before RCBT). This Pbto2 increase was associated with a significant mean increase in hemoglobin and hematocrit after RBCT (1.4 +/- 1.1 g/dL and 4.2% +/- 3.3%, respectively; both p < .001). Cerebral perfusion pressure, Sao2, and Fio2 were similar before and after RBCT. Among the 26 patients whose Pbto2 increased, the mean increase in Pbto2 was 5.1 +/- 9.4 mm Hg or a 49% mean increase (p < .01). CONCLUSIONS: RBCT is associated with an increase in Pbto2 in most patients with subarachnoid hemorrhage or traumatic brain injury. This mean increase appears to be independent of cerebral perfusion pressure, Sao2, and Fio2. Further study is required to determine why Pbto2 decreases in some patients after RBCT. PMID- 15891344 TI - Effects of sufentanil or ketamine administered in target-controlled infusion on the cerebral hemodynamics of severely brain-injured patients. AB - OBJECTIVE: The manual injection of a bolus of opioid in patients with brain injury induces an increase in intracranial pressure related to a decrease in mean arterial pressure. Such an effect has not been observed with the use of ketamine. The use of target-controlled infusion would minimize or suppress this adverse effect of opioid. This study evaluated the effects of an increase in plasma concentrations of sufentanil or ketamine administered by target-controlled infusion on cerebral hemodynamics. DESIGN: Prospective, randomized study. SETTING: Intensive care unit in a trauma center. PATIENTS: Thirty patients with severe traumatic brain injury. INTERVENTIONS: Patients were assigned to receive sedation consisting of sufentanil-midazolam or ketamine-midazolam using target controlled infusion. Twenty-four hours after the onset of sedation, the target concentrations of sufentanil or ketamine were doubled for 15 mins. Blood samples were collected to determine the actual plasma concentration of sufentanil and ketamine, before and 15 mins after concentration change. MEASUREMENTS AND MAIN RESULTS: The baseline values of intracranial pressure and cerebral perfusion pressure were similar in both groups. The two-fold increase in drug concentrations did not involve a significant change for intracranial pressure, cerebral perfusion pressure, and mean velocity of middle cerebral artery in both the ketamine and the sufentanil groups. The measured plasma concentrations of sufentanil and ketamine were 0.4 +/- 0.2 ng/mL and 2.6 +/- 2.2 mug/mL, respectively, before the increase in concentrations and 0.7 +/- 0.4 ng/mL and 5.5 +/- 3.8 mug/mL after. CONCLUSIONS: The present study shows that the increase in sufentanil or ketamine plasma concentrations using a target-controlled infusion is not associated with adverse effects on cerebral hemodynamics in patients with severe brain injury. The use of target-controlled infusion could be of interest in the management of severely brain-injured patients. However, there is a need for specific pharmacokinetic models designed for intensive care unit patients. PMID- 15891345 TI - Physiologic comparison between conventional mechanical ventilation and transtracheal open ventilation in acute traumatic quadriplegic patients. AB - OBJECTIVE: To evaluate the efficacy of mechanical ventilation administered through a small-bore, uncuffed tracheotomy tube, so-called transtracheal open ventilation (TOV), in comparison with conventional mechanical ventilation via a cuffed tracheal tube (endotracheal invasive ventilation, EIV). DESIGN: Physiologic study. SETTING: Intensive care unit of a referral trauma center. PATIENTS: Ten acute quadriplegic patients. INTERVENTIONS: In acute quadriplegic patients receiving EIV, TOV was subsequently applied via an uncuffed, small-bore tube (internal diameter of 4 or 5 mm). MEASUREMENTS AND MAIN RESULTS: Compared with EIV, arterial blood gases were not significantly different after 1 hr of TOV (Pao2/Fio2, 222.8 +/- 60.9 vs. 218.5 +/- 60.3; Paco2, 37.8 +/- 7.1 torr [5.04 +/- 0.95 kPa] vs. 35.5 +/- 6.8 torr [4.73 +/- 0.91 kPa], for EIV and TOV, respectively). Respiratory rate (19.5 +/- 4.7 vs. 19.6 +/- 5 breaths/min) and inspiratory effort (pressure-time product of esophageal pressure during a 1-min period, 125.9 +/- 48.4 vs. 112.8 +/- 36.4 cm H2O.sec.min) were also no different between the two modes. After 24 hrs of TOV, compared with EIV and TOV after 1 hr, respiratory rate and arterial blood gases remained stable, and the pressure-time product of esophageal pressure during a 1-min period was slightly, but significantly, reduced (83.5 +/- 16.6 cm H2O.sec.min, p < .05). CONCLUSIONS: In acute quadriplegic patients receiving mechanical ventilation, TOV was as effective as EIV in providing ventilatory support. PMID- 15891346 TI - Let us use the pulmonary artery catheter correctly and only when we need it. AB - OBJECTIVE: To clarify the issues related to the use of the pulmonary artery catheter within a rational clinical perspective. RESULTS: Barriers include a) increased patient risk of pulmonary artery catheter placement; b) ability to measure similar variables via central venous catheterization, echocardiography, or other less invasive techniques; c) increased cost; d) inaccurate measurements; e) incorrect interpretation and application of pulmonary artery catheter-derived variables; and f) lack of proven benefit of pulmonary artery catheter use in the overall management of patients. INTERPRETATION: a) The risks are mainly due to insertion of a central catheter, not a pulmonary artery catheter; b) continuous monitoring of left ventricular filling pressures, pulmonary vascular pressures, and mixed venous oxygen saturation is a unique feature; c) additional costs are minimal relative to the cost of intensive care; d) measurement errors require ongoing programmatic educational efforts; e) pulmonary artery catheter-derived data need to be used within the context of a defined treatment protocol; and f) no monitoring device, no matter how simple or sophisticated, will improve patient centered outcomes unless coupled with a treatment that, itself, improves outcome. CONCLUSION: A treatment protocol for the use of pulmonary artery catheter-derived variables is proposed that could serve as a basis for a prospective clinical trial. PMID- 15891347 TI - Another point of view: no swan song for the pulmonary artery catheter. PMID- 15891348 TI - Intestinal permeability and systemic infections in critically ill patients: effect of glutamine. AB - OBJECTIVE: This article provides a critical review of the evidence indicating that an increase in intestinal permeability is associated with the installation of bacteremia, sepsis, and the multiple organ failure syndrome and that glutamine in pharmacologic doses reduces the acute increase of intestinal permeability and the infection frequency in critically ill patients. DATA SOURCE: All studies published until December 2004 about intestinal permeability, bacterial translocation, and glutamine were located by search of PubMed and Web of Science. The reference lists of review articles and primary publications were also examined to identify references not detected in the computer search. STUDY SELECTION: Clinical and experimental studies investigating the correlation between intestinal permeability, bacterial translocation, and frequency of infections, associated or not with the effect of glutamine administration. DATA EXTRACTION: Information regarding patient population, experimental design, glutamine doses and routes of administration, nutritional therapy prescribed, methods used to assess intestinal permeability, metabolic variables, and the frequency of infections were obtained from the primary literature. DATA SYNTHESIS: Intestinal permeability is increased in critically ill patients. The results have not always been consistent, but the studies whose results support the association between intestinal permeability and systemic infections have had better design and more appropriate controls. The administration of glutamine by the intravenous or oral route and at the doses recommended before or immediately after surgery, burns, or the administration of parenteral nutrition has a protective effect that prevents or reduces the intensity of the increase in intestinal permeability. Glutamine reduces the frequency of systemic infections and may also reduce the translocation of intestinal bacteria and toxins, but this has not been demonstrated. CONCLUSIONS: Glutamine administration improves the prognosis of critically ill patients presumably by maintaining the physiologic intestinal barrier and by reducing the frequency of infections. PMID- 15891349 TI - The long-distance tertiary air transfer and care of tsunami victims: injury pattern and microbiological and psychological aspects. AB - OBJECTIVE: On December 26, 2004, a giant earthquake shocked Southeast Asia, triggering deadly flood waves (tsunami) across the Indian Ocean. More than 310,000 people have been reported dead and millions left destitute. Shortly thereafter, European governments organized airborne home transfer of most severely injured tourists using MedEvac aircraft. On arrival, patients were distributed to various medical centers. One cohort of the severely injured was admitted to the Cologne-Merheim Medical Center (Germany) for further surgical and intensive care treatment. The objective of this report was to characterize typical injury patterns along with microbiological findings and psychoemotional aspects unique to the tsunami disaster. DESIGN: Observational study. SETTING: Adult intensive care unit of a university hospital. PATIENTS: Seventeen severely injured tsunami victims were screened on arrival for characteristic injury patterns. In parallel, multifocal microbiological assessment was performed to identify pathogens responsible for high-level wound contamination. INTERVENTIONS: Standard clinical management. MEASUREMENTS AND MAIN RESULTS: The predominant pattern of injury comprised multiple large-scale soft-tissue wounds (range, 2 x 3 to 60 x 60 cm) of lower extremities (88%), upper extremities (29%), and head (18%). Additional injuries included thoracic trauma with hemopneumothorax and serial rib fractures (41%) and peripheral bone fractures (47%). A major problem associated with wound management was significant contamination. Microbiological assessment identified a variety of common (Pseudomonas 54%, Enterobacteriae 36%, Aeromonas spp. 27%) but also uncommon isolates that were often multiply resistant (multiply resistant Acinetobacter and extended-spectrum beta-lactamase-positive Escherichia coli, 18% each). Upper respiratory tract specimens contained a high rate of multiply resistant Acinetobacter species but also methicillin-resistant Staphylococcus aureus, Aeromonas hydrophilia, Pseudomonas species, and Candida albicans. Apart from these findings, all patients displayed severe signs of posttraumatic stress response. CONCLUSIONS: Individuals who survived their initial injuries and who were evacuated to Europe had traumatic injuries to head, chest, and limbs that were often contaminated with highly resistant bacteria. PMID- 15891350 TI - Mechanical ventilation in ARDS: One size does not fit all. PMID- 15891351 TI - Positive clinical impact of low tidal volume strategy. PMID- 15891352 TI - Coupling quality improvement with quality measurement in the intensive care unit. PMID- 15891353 TI - "If you cannot measure it, you cannot improve it". PMID- 15891354 TI - Cross-transmission in the intensive care unit: one piece of the puzzle. PMID- 15891355 TI - Optimal outcome in patients with community-acquired pneumonia: Take the main course before the dessert. PMID- 15891356 TI - Organizational factors affect human resuscitation: the role of simulation in resuscitation research. PMID- 15891357 TI - Sick? Or, not sick? PMID- 15891358 TI - Elevated cytokines in pneumococcal meningitis: Chicken or egg? PMID- 15891359 TI - Hypercapnic acidosis is mostly good for critically ill children: Also after cardiopulmonary bypass? PMID- 15891360 TI - NH2 terminal pro-brain natriuretic peptide in cardiovascular dysfunction and septic shock. PMID- 15891361 TI - Going on and on with NO? PMID- 15891362 TI - Tracheal access: on the cusp of a decision? PMID- 15891363 TI - Is cell death a prerequisite for cardiac dysfunction during sepsis? PMID- 15891364 TI - Ketamine in critical illness: another no-NO agent? PMID- 15891365 TI - Free radical scavenging as a therapeutic strategy for bacteremia. PMID- 15891366 TI - Bacterial translocation in hemorrhagic shock: Are we any wiser? PMID- 15891367 TI - Bacterial translocation and intestinal neutrophil lipid peroxidation in a hemorrhagic shock model--Rat race or rat trap? PMID- 15891368 TI - Improving the number and condition of donor lungs for transplantation. PMID- 15891369 TI - Spontaneous breathing during mechanical ventilation. PMID- 15891370 TI - Who needs blood? PMID- 15891371 TI - Target-controlled infusion for sedation of traumatic brain-injured patients: role uncertain. PMID- 15891372 TI - Transtracheal open ventilation and respiratory failure: The "missing link" between invasive and noninvasive approaches? PMID- 15891373 TI - Can glutamine turn off the motor that drives systemic inflammation? PMID- 15891374 TI - Tsunami disaster: a report from the front. PMID- 15891375 TI - Tsunami disaster and infection: Beware what pathogens the transport delivers to your intensive care unit! PMID- 15891377 TI - A lowered incidence of severe encephalopathy and cardiovascular dysfunction. PMID- 15891379 TI - A critical appraisal of the guidelines for the management of pediatric and neonatal patients with septic shock. PMID- 15891381 TI - Human albumin in critically ill patients. PMID- 15891383 TI - On the evaluation of the ISTH score for overt disseminated intravascular coagulation. PMID- 15891386 TI - Enhancing reverse cholesterol transport: the case for phosphatidylcholine therapy. PMID- 15891387 TI - Assembly of high density lipoprotein by the ABCA1/apolipoprotein pathway. AB - PURPOSE OF REVIEW: Mammalian somatic cells do not catabolize cholesterol and therefore need to export it for sterol homeostasis at the levels of cells and whole bodies. This mechanism may reduce intracellularly accumulated cholesterol in excess, and thereby would contribute to the prevention or cure of the initial stage of atherosclerotic vascular lesions. RECENT FINDINGS: HDL is thought to play a main role in this reaction on the basis of epidemiological evidence and in vitro experimental data. Two independent mechanisms have been identified for this reaction. One is non-specific diffusion-mediated cholesterol 'efflux' from the cell surface, and cholesterol is trapped by various extracellular acceptors including lipoproteins. Extracellular cholesterol esterification on HDL provides a driving force for the net removal of cell cholesterol, and some cellular factors may enhance this reaction. The other mechanism is an apolipoprotein mediated process to generate HDL by removing cellular phospholipid and cholesterol. This reaction is mediated by a membrane protein ABCA1, and lipid free or lipid-poor helical apolipoproteins recruit cellular phospholipid and cholesterol to assemble HDL particles. The reaction is composed of two elements: the assembly of HDL particles with phospholipid by apolipoprotein, and cholesterol enrichment in HDL. ABCA1 is essential for the former step, and the latter step requires further intracellular events. SUMMARY: ABCA1 is a rate limiting factor of HDL assembly and is regulated by transcriptional factors and posttranscriptional factors. Posttranscriptional regulation of ABCA1 involves the modulation of its calpain-mediated degradation. PMID- 15891388 TI - Intestinal lipoprotein assembly. AB - PURPOSE OF REVIEW: The assembly of intestinal lipoproteins is critical for the transport of fat and fat-soluble vitamins. In this review we propose a nomenclature for these lipoproteins and have summarized recent data about their intracellular assembly and factors that modulate their secretion. RECENT FINDINGS: The assembly and secretion of intestinal lipoproteins increases with the augmented synthesis of apoB, apoAIV and lipids. Chylomicron assembly begins with the formation of primordial, phospholipid-rich particles in the membrane, and their conversion to large chylomicrons occurs in the lumen of the smooth endoplasmic reticulum. Chylomicrons are transported from the endoplasmic reticulum via specialized vesicles to the Golgi for secretion. The identification of genetic mutations in chylomicron retention disease indicates that Sar1b may play a critical role in this process. In addition to chylomicron assembly, intestinal cells have been shown to transport dietary cholesterol via apoB independent pathways, such as efflux. SUMMARY: Understanding the mechanisms involved in the intracellular transport of chylomicrons and chylomicron independent secretion pathways are expected to be the next frontiers in the field of intestinal lipoprotein assembly and secretion. PMID- 15891389 TI - Apolipoprotein structure and dynamics. AB - PURPOSE OF REVIEW: This review highlights recent advances in structural studies of exchangeable human apolipoproteins and the insights they provide into lipoprotein action in cardiovascular and amyloid diseases. RECENT FINDINGS: The high-resolution X-ray crystal structure of free apoA-II reveals a parallel helical array that may represent other lipid-poor apolipoproteins, and the structure in complex with detergent substantiates the belt model for the protein arrangement on lipoproteins. Nuclear magnetic resonance structures of apolipoprotein-detergent complexes show a repertoire of curved helical conformations, suggesting multiple helical arrangements on the lipid. Low resolution spectroscopic analyses, interface studies and molecular modeling provide new insights into the 'hinge-domain' mechanism of apolipoprotein adaptation at variable lipoprotein surfaces. A kinetic mechanism for lipoprotein stabilization is proposed. SUMMARY: Cumulative evidence supports the belt model that provides a general structural basis for understanding the molecular mechanisms of functional apolipoprotein reactions, such as binding to lipoprotein receptors, lipid transporters, and the activation of lipophilic enzymes. However, the detailed protein and lipid conformations on lipoproteins and the underlying molecular interactions are unclear. New insights will hopefully emerge once the first detailed lipoprotein structure is solved. PMID- 15891390 TI - Apolipoprotein structural organization in high density lipoproteins: belts, bundles, hinges and hairpins. AB - PURPOSE OF REVIEW: To summarize recent advances towards an understanding of the three-dimensional structures of the apolipoprotein components of HDL with a specific focus on high resolution models of apolipoprotein A-I. RECENT FINDINGS: Since the primary sequence was first reported, various models have been advanced for the structure of apolipoprotein A-I, the major protein constituent of HDL, in its lipid-free and lipid-bound forms. Unfortunately, the generation of experimental data capable of distinguishing among the competing models has lagged far behind. However, recent experimental strategies, including X-ray crystallography, applications of resonance energy transfer and mass spectrometry, have combined with sophisticated theoretical approaches to develop three dimensional structural models of apolipoprotein A-I with previously unavailable resolution. SUMMARY: The recent synergy of sophisticated computer modeling techniques with hard experimental data has generated new models for apolipoprotein A-I in certain subclasses of HDL produced in vitro. The challenge now is to adapt and test these models in the more complex forms of HDL isolated directly from human plasma. PMID- 15891391 TI - The role of the kidney in lipid metabolism. AB - PURPOSE OF REVIEW: Cellular uptake of plasma lipids is to a large extent mediated by specific membrane-associated proteins that recognize lipid-protein complexes. In the kidney, the apical surface of proximal tubules has a high capacity for receptor-mediated uptake of filtered lipid-binding plasma proteins. We describe the renal receptor system and its role in lipid metabolism in health and disease, and discuss the general effect of the diseased kidney on lipid metabolism. RECENT FINDINGS: Megalin and cubilin are receptors in the proximal tubules. An accumulating number of lipid-binding and regulating proteins (e.g. albumin, apolipoprotein A-I and leptin) have been identified as ligands, suggesting that their receptors may directly take up lipids in the proximal tubules and indirectly affect plasma and tissue lipid metabolism. Recently, the amnionless protein was shown to be essential for the membrane association and trafficking of cubilin. SUMMARY: The kidney has a high capacity for uptake of lipid-binding proteins and lipid-regulating hormones via the megalin and cubilin/amnionless protein receptors. Although the glomerular filtration barrier prevents access of the large lipoprotein particles to the proximal tubules, the receptors may be exposed to lipids bound to filtered lipid-binding proteins not associated to lipoprotein particles. Renal filtration and receptor-mediated uptake of lipid binding and lipid-regulating proteins may therefore influence overall lipid metabolism. The pathological mechanisms causing the pronounced atherosclerosis promoting effect of uremia may involve impairment of this clearance pathway. PMID- 15891392 TI - Scavenger receptor BI and ATP-binding cassette transporter A1 in reverse cholesterol transport and atherosclerosis. AB - PURPOSE OF REVIEW: The appearance of scavenger receptor class B type I (SR-BI) and ATP-binding cassette transporter A1 (ABCA1) in macrophages and liver implicates these transporters in different stages of reverse cholesterol transport. This review focuses on the role of SR-BI and ABCA1 in reverse cholesterol transport in the context of atherosclerotic lesion development. RECENT FINDINGS: Recent studies indicate that hepatic expression of ABCA1 and SR BI is important for the generation of nascent HDL and the delivery of HDL cholesteryl esters to the liver, respectively. Although macrophage SR-BI and ABCA1 do not contribute significantly to circulating HDL levels, the perpetual cycle of HDL lipidation and delipidation by the liver ensures the availability of acceptors for cholesterol efflux that maintain cholesterol homeostasis in arterial macrophages, thereby reducing atherogenesis. In addition to its established role in the selective uptake of HDL cholesteryl esters, there is now evidence that hepatic SR-BI facilitates postprandial lipid metabolism, and that hepatic secretion of VLDL is dependent on ABCA1-mediated nascent HDL formation. Thus, remnant and HDL metabolism are more intimately intertwined in hepatic lipid metabolism than has previously been appreciated. SUMMARY: Recent advances in the understanding of the role of ABCA1 and SR-BI in HDL metabolism and their atheroprotective properties indicate the significant potential of modulating ABCA1 and SR-BI expression in both arterial wall macrophages and the liver for the treatment of atherosclerotic coronary artery disease. PMID- 15891393 TI - The mammalian target of rapamycin signaling network and gene regulation. AB - PURPOSE OF REVIEW: The mammalian target of rapamycin (mTOR) is a key integrator of signals from nutrients, energy and insulin. TOR is a protein kinase originally identified in yeast by the genetic selection of rapamycin-resistant mutants. Over the past decade mTOR research has progressed dramatically. Although mTOR is known as a controller of messenger RNA cap-dependent translation initiation, new advances implicate mTOR in the regulation of ribosomal protein gene transcription. The aim of this review is to highlight recent findings on mTOR regulatory networks, focusing on articles published from December 2003 to December 2004. RECENT FINDINGS: mTOR was recently knocked out in mice; the embryonic lethal phenotype demonstrates a critical role of mTOR in early embryo development. Intriguingly, the homozygous deletion of ribosomal protein S6 kinase 1 (S6K1), an mTOR target, in mice results in hypoinsulinemia and glucose intolerance. Despite elevated levels of plasma free fatty acids, S6K1 knockout mice are protected from the metabolic syndrome, indicating a role of S6K1 in glucose homeostasis. Current research indicates that mTOR integrates input from multiple upstream pathways, including insulin, growth factors, nutrients, mitogens and energy. Furthermore, the discovery of mTOR binding partners adds to the intricacies of mTOR as a master switch in cell signaling. SUMMARY: Rapamycin, an mTOR inhibitor, has emerged as an immunosuppressive and antiproliferative drug, and is considered a novel antitumor agent. A better understanding of mTOR signaling would enhance the clinical usefulness of rapamycin and inform consideration of mTOR as a target for the development of new therapies. PMID- 15891394 TI - Evolution and mechanism of apolipoprotein B-containing lipoprotein assembly. AB - PURPOSE OF REVIEW: Apolipoprotein B-containing lipoprotein assembly and secretion is critical for lipid absorption and triglyceride homeostasis, and plays a role in atherogenesis and the pathobiology of type 2 diabetes and obesity. This review highlights recent insights into the evolutionary, structural, and cell biology of hepatic and intestinal pathways for lipid mobilization, and the mechanisms and regulation of lipoprotein assembly and secretion. RECENT FINDINGS: Until recently it was assumed that microsomal triglyceride transfer protein-dependent apolipoprotein B-containing lipoprotein assembly was a unique adaptation associated with vertebrate lipid homeostasis. However, it is now clear that microsomal triglyceride transfer protein (MTP) exists in species whose last common ancestor diverged over 550 million years ago. In its long evolutionary history, the MTP gene has given rise to a series of paralogous lipid transport proteins, all of which require MTP for their biogenesis. During its evolution, MTP has acquired new functions, enabling it to participate in a disparate array of lipid mobilization and transport pathways, ranging from primitive lipoprotein assembly to antigenic lipid presentation. In addition to the complex and multifunctional role of MTP in apolipoprotein B assembly, other factors responsible for the generation of secretion-coupled lipids and the modulation of apolipoprotein B production are emerging. SUMMARY: The phylogenic dissection of MTP and apolipoprotein B function, coupled with ongoing structural and biochemical analyses, provide significant insights into the mechanisms of lipid mobilization and secretion. Some of these factors and processes may be targeted therapeutically to modulate the quantitative and qualitative aspects of apolipoprotein B production. PMID- 15891395 TI - Lipolysis: pathway under construction. AB - PURPOSE OF REVIEW: The lipolytic catabolism of stored fat in adipose tissue supplies tissues with fatty acids as metabolites and energy substrates during times of food deprivation. This review focuses on the function of recently discovered enzymes in adipose tissue lipolysis and fatty acid mobilization. RECENT FINDINGS: The characterization of hormone-sensitive lipase-deficient mice provided compelling evidence that hormone-sensitive lipase is not uniquely responsible for the hydrolysis of triacylglycerols and diacylglycerols of stored fat. Recently, three different laboratories independently discovered a novel enzyme that also acts in this capacity. We named the enzyme 'adipose triglyceride lipase' in accordance with its predominant expression in adipose tissue, its high substrate specificity for triacylglycerols, and its function in the lipolytic mobilization of fatty acids. Two other research groups showed that adipose triglyceride lipase (named desnutrin and Ca-independent phospholipase A2zeta, respectively) is regulated by the nutritional status and that it might exert acyl transacylase activity in addition to its activity as triacylglycerol hydrolase. Adipose triglyceride lipase represents a novel type of 'patatin domain containing' triacylglycerol hydrolase that is more closely related to plant lipases than to other known mammalian metabolic triacylglycerol hydrolases. SUMMARY: Although the regulation of adipose triglyceride lipase and its physiological function remain to be determined in mouse lines that lack or overexpress the enzyme, present data permit the conclusion that adipose triglyceride lipase is involved in the cellular mobilization of fatty acids, and they require a revision of the concept that hormone-sensitive lipase is the only enzyme involved in the lipolysis of adipose tissue triglycerides. PMID- 15891396 TI - Secretory phospholipase A2 enzymes in atherogenesis. AB - PURPOSE OF REVIEW: Immunohistochemistry studies have confirmed the presence of group IIA, group V and group X secretory phospholipase A2 in human or mouse atherosclerotic lesions. The possibility that secretory phospholipase A2 plays a role in the pathophysiology of atherosclerosis (and is not merely a marker for localized inflammation) has been substantiated by a number of recent in-vitro and in-vivo studies. RECENT FINDINGS: A mouse strain with a targeted deletion of group V secretory phospholipase A2 has been developed. Peritoneal macrophages from these mice have significantly blunted eicosanoid generation in response to zymosan, providing the first direct evidence that a secretory phospholipase A2 plays a role in stimulation-induced arachidonic acid production in vivo. A recent in-vitro study indicated that de novo synthesized groups IIA and X secretory phospholipase A2 can mediate arachidonic acid release intracellularly, without the requirement for previous secretion from cells, as was previously thought. Several studies support the previously proposed model that secretory phospholipase A2 hydrolysis generates pro-atherogenic LDL. These data, coupled with the finding that macrophage-specific expression of human group IIA secretory phospholipase A2 promotes atherosclerotic lipid deposition in mice, draw attention to secretory phospholipase A2 as an attractive target for the treatment of atherosclerotic disease. SUMMARY: Secretory phospholipase A2 activity in the arterial intima has the potential to amplify atherogenic processes by liberating potent pro-inflammatory lipid mediators and by generating pro-atherogenic LDL. Future in-vivo studies will aid in defining the mechanism(s) that underlie the pro-atherosclerotic effects of secretory phospholipase A2. PMID- 15891397 TI - High-density lipoproteins as therapeutic targets. AB - PURPOSE OF REVIEW: The concentration of cholesterol in HDL is an inverse predictor of future cardiovascular disease, with evidence mounting that therapies that increase HDL concentration are antiatherogenic. The best known antiatherogenic function of HDL particles relates to their ability to promote the efflux of cholesterol from cells. However, they also have antioxidant, antiinflammatory and antithrombotic properties. RECENT FINDINGS: The past year has seen the publication of several papers that highlight a potential major protective role of HDL in states of acute inflammation. Papers showing extremely promising results using novel inhibitors of cholesteryl ester transfer protein as HDL-raising agents have also appeared. Finally, the discovery that ATP-binding cassette transporter G1 (ABCG1) transports cell cholesterol to large HDL particles in the extracellular space has largely reconciled apparent inconsistencies between basic research indicating that small, pre-beta-migrating HDL particles are the antiatherogenic components of HDL and epidemiological research that implicates larger HDL particles as the protective fraction. SUMMARY: The finding that ABCG1 promotes the efflux of cholesterol from cells to large HDL particles also provides powerful circumstantial evidence that cholesteryl ester transfer protein inhibition (which increases HDL size) may enhance, rather than reduce, cholesterol efflux, and thus inhibit the development of atherosclerosis. PMID- 15891398 TI - Bibliography. Current world literature. Lipid metabolism. PMID- 15891399 TI - Genetics and molecular biology. PMID- 15891400 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 15891402 TI - Multiple sclerosis--novel insights and new therapeutic strategies. AB - PURPOSE OF REVIEW: This review focuses on novel aspects of the pathogenesis and advances in the therapy of multiple sclerosis (MS). RECENT FINDINGS: Recent observations suggest that early lesion development in MS may start in some forms with oligodendrocyte death and that inflammation appears as a secondary phenomenon only. The lack of sufficient remyelination in MS may be the result of a disturbed function of basic helix-loop-helix transcription factors. Clinically the identification of patients with a clinically isolated syndrome at high risk to develop clinically definite MS remains difficult; the predictive value of serum antibodies against myelin proteins remains controversial. The role of neutralizing antibodies in interferon therapy is discussed. New therapeutic approaches in MS are emerging. SUMMARY: The existing view on the pathogenesis of MS is still changing. The original assumption that cell-mediated demyelination is the key event in lesion development dictating clinical disability is critically reviewed and alternative pathways have been suggested. Oligodendrocyte death, axonal loss, the role of CD8 T lymphocytes, T regulatory cells, and B lymphocytes have come into the focus of newly evolving concepts in MS pathogenesis. A deepened understanding of the immunopathogenesis of this disease translates into innovative therapeutic approaches, such as blockade of alpha4 integrins by a humanized monoclonal antibody. In various animal models cell-replacement strategies yield promising results; however, turning these findings into an effective therapy in MS patients has a long way to go. PMID- 15891403 TI - The spectrum of multiple sclerosis: new lessons from pathology. AB - PURPOSE OF REVIEW: This review will focus on recent developments in multiple sclerosis (MS) pathology with particular emphasis on the patterns and mediators of lesion formation in MS, the mechanisms of oligodendrocyte and axon damage and the magnetic resonance imaging-pathological correlation of MS lesions. RECENT STUDIES: The inflammatory cascade in the MS plaque has been characterized in more detail, and other factors such as hypoxia-like injury or excitotoxicity, besides immunological effector mechanisms, have been found to play a role in MS pathogenesis. Cortical demyelination and mechanisms of neuroaxonal damage are discussed in detail. The radiological correlate of basic histopathologic findings is being approached with quantitative methods. Similar quantitative approaches are used in MS-gene expression studies that compare patterns of gene expression in different lesion areas. SUMMARY: These studies will lead to better understanding of the pathogenesis of MS lesions and will hopefully identify new therapeutic targets to modulate inflammation, support remyelination and protect axons and neurons in MS. PMID- 15891404 TI - Pathogenesis of multiple sclerosis. AB - PURPOSE OF REVIEW: The aim of this article is to describe recent observations regarding the basis for the initiation and disease evolution of multiple sclerosis. RECENT FINDINGS: A current debate is where and what initiates the neuroinflammatory reaction that characterizes the acute multiple sclerosis lesion. Immune sensitization to neural antigens could develop within the systemic compartment consequent to exposure to cross-reacting, possibly viral derived, peptides (molecular mimicry). Although CD4 T cells are considered central to initiating central nervous system inflammation, the actual extent and specificity of tissue injury reflects the array of adaptive (CD8 T cells and antibody) and innate (microglia/macrophages) immune constituents present in the lesions. Neuropathologic studies indicate that lethal changes in neural cells (oligodendrocytes) could also be the initiating event, reflecting as yet unidentified acquired insults (e.g. exogenous virus or reactivated endogenous retrovirus) or intrinsic abnormalities ('neurodegenerative' hypothesis). Recurrence or persistence of the disease process can reflect events occurring at multiple sites including expansion of the immune repertoire in response to neural antigens transported to regional lymph nodes (determinant spreading), especially if immune regulatory mechanisms are defective; alterations in blood-brain barrier properties consequent to initial cellular transmigration; and participation of endogenous (microglia, astrocytes) or long lived infiltrating cells (macrophages, B cells in ectopic germinal centers) in regulating and effecting immune functions within the central nervous system. Accumulating neurologic deficit reflects the balance between injury and repair; the latter also being negatively or positively (trophic support and clearance of tissue debris) impacted by inflammatory processes. SUMMARY: Understanding the full spectrum of multiple sclerosis presents a continuing challenge for both immunology and neurobiology. PMID- 15891405 TI - Genomics, proteomics, metabolomics: what is in a word for multiple sclerosis? AB - PURPOSE OF REVIEW: Multiple sclerosis (MS) is the most common chronic inflammatory neurological disease. Despite major advances the aetiology of this disease it is still not completely understood. In the post-genome era, advances in global screening technologies offer an opportunity to accelerate the search of new pathological pathways and to identify new therapeutic targets. Some recent publications using novel global screening methods at the genome, transcriptome, proteome and metabolome levels are discussed. RECENT FINDINGS: The genetic association of susceptibility to MS with loci outside the MHC has been reconfirmed. Evidence of parent-of-origin and seasonal effects on disease susceptibility add further complexity to the genetics of MS. The search for MS susceptibility genes continues using the candidate-gene approach as well as large scale single-nucleotide-polymorphism association studies and novel cross-species synteny analysis. Genome-wide expression profiling using microarrays produced numerous therapeutic targets and is progressing towards profiling of rare cells. Advances in classical proteomics methods paved the way to new initiatives aiming at determining the proteome of the nervous system in normal and diseased states. Although progress is still slow, array-based methods are making an impact on the MS field. SUMMARY: The complexity of MS is clearly reflected in the latest findings using global profiling methods. Nevertheless, these new technologies are confirming some of the basic aspects of the disease pathophysiology, i.e. its polygenicity, the central role of neuroinflammation and the emerging neurodegenerative processes. These data are primarily the results of genomic approaches, yet promising attempts are also made using proteomics and metabolomics. PMID- 15891406 TI - Promoting repair in multiple sclerosis: problems and prospects. AB - PURPOSE OF REVIEW: Despite recent progress in treating the inflammatory component of multiple sclerosis, current therapies have no clear impact on progression of disability, which closely relates to tissue (myelin and axon) injury. Many scientists now focus their efforts on elucidating the mechanisms that lead to tissue injury, and on developing new strategies for tissue repair. We review recent breakthroughs in this field and discuss their putative applications to therapy. RECENT FINDINGS: Several hypotheses have been raised to explain the failure of remyelination, including depletion of remyelinating cells, quiescence of oligodendrocyte precursor cells and axonal inhibitory signals. Success in remyelination therapy may be achieved either by enhancing endogenous repair or by grafting exogenous remyelinating cells. Several neurotrophic factors have been shown to enhance endogenous remyelination, and many immature cells have been shown to induce efficient exogenous remyelination in animal models. Although effective remyelination probably represents the best way to prevent neurodegeneration, several alternative neuroprotective strategies are emerging. Statins, cyclins and immunophilin ligands are orally available immunomodulatory agents that may protect neurones. Other promising possibilities include the modulation of excitotoxicity, nitric oxide synthesis, or cationic channels. SUMMARY: Despite the increasing number of putative therapeutic targets, no treatment to achieve remyelination or neuroprotection has yielded positive clinical results in humans. Forging a link between basic biology and treatment of patients will require us to overcome several challenges, including assessment of efficacy of repair, improving tolerance to and delivery of neurotrophic factors, and better defining the indications for and limitations of transplantation. PMID- 15891407 TI - Clinical trials in multiple sclerosis: methodological issues. AB - PURPOSE OF REVIEW: The availability of partially effective immunomodulatory and immunosuppressive treatments for relapsing multiple sclerosis (MS) opens important ethical, methodological and practical issues in the design and conduct of new clinical trials in these patients. RECENT FINDINGS: The recommendation of the National Health Authorities to prioritize phase III clinical trials using placebo arm raises ethical questions. In addition, patients are reluctant to be involved in such trials. Alternative clinical trial designs will be discussed. Relapses and active lesions are accepted measures of disease activity; new/enlarging T2 lesions and/or enhancing lesions are accepted surrogate markers of disease activity in phase II clinical trials. On the contrary, there are no accepted magnetic resonance imaging (MRI) surrogate markers of disease progression and also the clinical measures to monitor the degenerative aspects of the disease are not without important limitations. New scales of impairment, disability and quality of life will be reviewed extensively. We will also focus on the value of modern and quantitative MRI techniques, which hold substantial promise as tools to estimate the extent of MS-related irreversible tissue loss. SUMMARY: The use of an active comparator in a superior clinical-trial design is becoming an attractive option for testing the efficacy of new drugs in relapsing MS. At present there are no fully reliable and sensitive clinical markers of the accumulation of irreversible tissue damage in MS. Although additional extensive application in longitudinal studies is needed, modern MRI techniques are promising tools to monitor the neurodegenerative aspects of MS. PMID- 15891408 TI - Current trials in multiple sclerosis: established evidence and future hopes. AB - PURPOSE OF REVIEW: The aim of the present report is to briefly review multiple sclerosis therapeutic trials published or presented in 2004 to provide an up-to date overview of the established evidence and new insights. RECENT FINDINGS: New data have come available that help us understand how currently approved disease modifying drugs can best be used. Nonetheless, their limited effectiveness - especially in progressive forms of multiple sclerosis - as well as the inconvenience and toxicity associated with their use, emphasize the need for new treatment strategies. A substantial number of reports on new emerging treatment modalities were published in 2004, and one of these modalities was newly approved by the US Food and Drug Administration for the treatment of relapsing forms of multiple sclerosis. SUMMARY: Further advances have been made in the treatment of multiple sclerosis patients. On the one hand, we know better how and in whom to use existing medications. On the other hand, it is exciting to witness how increased insight in the pathophysiology of the disease and its symptoms has led to a series of new, innovative treatment modalities. PMID- 15891409 TI - Primary progressive multiple sclerosis. AB - PURPOSE OF REVIEW: The present article reviews the currently ongoing scientific debate on the specific characteristics of primary progressive multiple sclerosis. RECENT FINDINGS: The most important observations come from the studies using magnetic resonance imaging showing involvement of the normal-appearing brain tissue and also from the clinical and magnetic-resonance-imaging descriptions in longitudinal studies. SUMMARY: Progress in the diagnosis of primary progressive multiple sclerosis has been made. Long- and short-term natural history are now better known, which will allow the designing of clinical trials in the near future. Magnetic-resonance-imaging studies have demonstrated damage of the normal appearing brain tissue, which may explain in part the apparent clinical and radiological paradox, common to all clinical forms of multiple sclerosis but perhaps more evident in the primary progressive form. Preliminary results from exploratory trials seem to indicate that these patients should no longer be excluded from therapeutic trials. PMID- 15891410 TI - Neurorehabilitation in multiple sclerosis: foundations, facts and fiction. AB - PURPOSE OF REVIEW: This review of recent work in the area of neurorehabilitation of multiple sclerosis patients surveys progress and underscores the need for further work to evaluate the effectiveness of treatments. RECENT FINDINGS: Several recent review documents have summarized the current position regarding neurorehabilitation and symptomatic management in multiple sclerosis. They have highlighted the paucity of evidence underpinning current practice, thereby identifying the need for more scientifically sound studies in both neurorehabilitation and symptomatic treatment. However, as will be apparent from this review, there has been a welcome increase in studies evaluating both aspects of neurorehabilitation and specific areas such as the role of cannabinoids in spasticity and pain and new treatments for cognitive impairment. SUMMARY: Overall, there is an encouraging trend both in questioning our current practice and in designing more scientifically sound trials incorporating new and more appropriate outcome measures. There is, however, much more to be done before we are in a position to provide the expert, comprehensive, joined-up, care that is required to meet the complex, ever-changing needs of patients with multiple sclerosis. PMID- 15891411 TI - Chronic inflammatory demyelinating polyneuropathy--update on pathogenesis, diagnostic criteria and therapy. AB - PURPOSE OF REVIEW: Chronic inflammatory demyelinating polyneuropathy (CIDP) is a treatable but possibly underdiagnosed disorder of the peripheral nerve. This review covers the growing literature of the past years that deals with the pathogenesis, diagnostic criteria and treatment of CIDP. RECENT FINDINGS: The recent development of a biphasic animal model of experimental autoimmune neuritis may provide further insights into the pathogenesis of inflammatory demyelination of the peripheral nerve, such as in CIDP, and may allow the development of further innovative therapeutic strategies. In patients, the contribution of immune processes to the dysfunction in hereditary polyneuropathies and the association of hereditary neuropathy and CIDP has been described. Commonly used therapies remain corticosteroids, intravenous immunoglobulin and plasmapheresis; however, newer immunosuppressant approaches using mycophenolate mofetil or cyclosporin A, or immunomodulating therapies using monoclonal antibodies or interferons are presently under investigation. SUMMARY: The growing body of knowledge on the pathogenesis of CIDP and further diagnostic differentiation of subforms may help to develop more-effective therapies for CIDP in the next few years. PMID- 15891412 TI - Headache. PMID- 15891413 TI - New targets in the acute treatment of headache. AB - PURPOSE OF REVIEW: The aim of this article is to review recently identified targets for the acute treatment of primary headache disorders. RECENT FINDINGS: Calcitonin gene-related peptide (CGRP) receptor blockade has been shown to be an effective acute anti-migraine strategy and is a non-vasoconstrictor in terms of the mechanism of action. It is likely that direct blockade of CGRP release by inhibition of trigeminal nerves would be similarly effective in both migraine and cluster headache. Options for acute treatment based on preclinical work and initial clinical studies include: serotonin 5HT1F and 5HT1D receptor agonists, glutamate excitatory amino acid receptor antagonists, nitric oxide synthase inhibitors and adenosine A1 receptor agonists. Proof of principle studies with octreotide, a somatostatin receptor agonist, demonstrated it to be better than placebo in the acute treatment of cluster headache but not in the acute management of migraine. SUMMARY: The prospect of a non-vasoconstrictor acute migraine therapy offers a real opportunity to patients, and perhaps more importantly, provides a therapeutic rationale to plant migraine and cluster headache firmly in the brain as neurological problems. PMID- 15891414 TI - Preventive treatment of headaches. AB - PURPOSE OF REVIEW: To review recent advances in preventive headache treatment. RECENT FINDINGS: Migraine may be a progressive disorder. Aggressive treatment may stop progression. Propranolol, the beta-blocker, and the anticonvulsant topiramate are effective for migraine prevention. Feverfew, montelukast and acupuncture have not proven effective. SUMMARY: New drugs and other treatment strategies expand the spectrum of preventive migraine treatments. PMID- 15891415 TI - The role of imaging in the pathophysiology and diagnosis of headache. AB - PURPOSE OF REVIEW: Functional neuroimaging in headache patients has revolutionized our understanding of these syndromes and provided unique insights into some of the most common maladies in humans, suggesting that at least migraine and cluster headache are driven primarily from the brain. This review highlights new studies and recent advances in studying headache using neuroimaging. RECENT FINDINGS: Concerning the diagnostics of headache, an EFNS Task Force evaluated recently the usefulness of imaging procedures in non-acute headache patients on the basis of evidence from the literature and defined guidelines on when to use magnetic resonance imaging or computed tomography. Regarding the pathophysiology of primary headache syndromes, repeated and independent findings reinforce the crucial role for the brainstem in acute and probably also in chronic migraine, and the hypothalamic grey in several trigemino autonomic headaches. If further studies confirm these findings, a better understanding will be gained of where and how acute and preventive therapy can be targeted. SUMMARY: Given the rapid advances in functional neuroimaging, in particular newer techniques such as voxel-based morphometry and magnetic resonance spectrometry, functional imaging continues to play a significant role and opens new avenues in targeting the neural substrates in individual primary headache syndromes. PMID- 15891416 TI - Patent foramen ovale: paradoxical connection to migraine and stroke. AB - PURPOSE OF REVIEW: In this article we aim to elucidate the relationship between patent foramen ovale, cryptogenic stroke and migraine. RECENT FINDINGS: Small observational and case-control studies indicate that patients with cryptogenic stroke have a higher incidence of patent foramen ovale. Prospective trials could not show a higher stroke recurrence risk with isolated patent foramen ovale. The combination of patent foramen ovale and atrial septal aneurysm might carry a higher recurrence risk. Secondary prevention with acetylsalicylic acid is as effective as oral anticoagulation, but carries a lower bleeding risk. Whether patent foramen ovale closure prevents recurrent strokes is under investigation. Case-control studies and retrospective analyses indicate comorbidity between patent foramen ovale and migraine, in particular migraine with aura. Recent retrospective studies indicate a reduction in migraine frequency after patent foramen ovale closure (intended for stroke prevention). These studies, however, have major methodological limitations. Therefore patent foramen ovale closure cannot be recommended for the prevention of migraine with aura. SUMMARY: At present routine percutaneous closure of isolated patent foramen ovale cannot be recommended for patients with cryptogenic stroke. Patent foramen ovale closure should not be used for the prevention of migraine. PMID- 15891417 TI - Comorbidity of migraine. AB - PURPOSE OF REVIEW: Comorbidity refers to the greater than coincidental association of separate conditions in the same individuals. Historically, a number of conditions have been noted to be comorbid with migraine, notably psychiatric disorders (anxiety, depression, panic disorder), epilepsy, asthma, and some congenital heart defects. Migraine sufferers have increased medical costs overall compared with others of the same sex and age, even after considering the cost of specific migraine treatment. Thus, estimates of the burden of migraine often include the costs of conditions comorbid with it. RECENT FINDINGS: Conditions may be comorbid through a variety of mechanisms. Comorbidity may be an artifact of diagnostic uncertainty when symptom profiles overlap or when diagnosis is not based on objective markers. Comorbidity may arise due to unidirectional causality, such as migraine resulting in blood pressure changes due to headache-specific treatment. Finally, conditions may be comorbid because of shared genetic or other factors that increase the risk of both conditions. In such cases, understanding these shared risk factors may lead to greater understanding of the fundamental mechanisms of migraine. SUMMARY: In this article, we will review recent developments related to migraine comorbidity. We will emphasize findings related to the comorbidity of migraine with clinical and sub-clinical vascular brain lesions, congenital heart defects, coronary heart disease, psychiatric illness, and other pain conditions. PMID- 15891418 TI - Update on Susac's syndrome. AB - PURPOSE OF VIEW: We review recent developments in the clinical course and imaging modalities for Susac's syndrome, a clinical triad consisting of encephalopathy, branch retinal artery occlusions and sensorineural hearing loss. RECENT FINDINGS: Susac's syndrome has variable clinical presentations; recently described presentations include epileptic seizures and transient inverted vision. Advances in neuroradiology suggest that magnetic resonance imaging demonstrates distinctive patterns in the white and grey matter and in the leptomeninges. Reports have verified that Susac's syndrome is under-diagnosed because of its multisystem involvement and confusion with other imitating disorders (such as multiple sclerosis), and because of the fact that neuroradiologists are not acquainted with this syndrome. SUMMARY: The precise aetiology of Susac's syndrome is still unknown and many areas have not yet been explored. Magnetic resonance imaging is the neuroimaging study of choice. Findings include multiple small hyperintense foci on T2-weighted images and contrast enhancement in white and grey matter of both supratentorial and infratentorial structures, corpus callosum and, occasionally, leptomeninges. Callosal lesions typically involve the central fibres and are probably pathognomonic for Susac's syndrome. When assessing patients with unexplained encephalopathy involving white and grey matter, leptomeninges and corpus callosum, the findings of sensorinueral hearing loss or visual disturbances may yield important clues regarding the possibility of Susac's syndrome. PMID- 15891419 TI - Role of inflammation in neurodegenerative diseases. AB - PURPOSE OF REVIEW: Inflammation is a self-defensive reaction aimed at eliminating or neutralizing injurious stimuli, and restoring tissue integrity. In neurodegenerative diseases inflammation occurs as a local response driven by microglia, in the absence of leucocyte infiltration. Like peripheral inflammation, neuroinflammation may become a harmful process, and it is now widely accepted that it may contribute to the pathogenesis of many central nervous system disorders, including chronic neurodegenerative diseases. This review addresses some of the most recent advances in our understanding of neuroinflammation. RECENT FINDINGS: The presence of activated microglia surrounding amyloid plaques and increased levels of complement elements, cytokines, chemokines and free radicals support the existence of a self propagating toxic cycle and provide a rationale for anti-inflammatory approaches to prevent or delay neurodegeneration. Nonetheless, recent studies have provided evidence that chronic stimulation leads microglia to acquire an anti-inflammatory phenotype, characterized by activated morphology and induction of neuroprotective and immunoregulatory molecules. The causes and consequences of this atypical phenotype have just begun to be unravelled. SUMMARY: Although significant advances have been made in our knowledge of degenerative diseases, there remains controversy regarding whether neuroinflammation and microglial activation are beneficial or detrimental. Strategies aimed at both preventing and boosting microglial activation are presently under investigation, and these studies might reveal new potentially effective treatments for these neurological disorders. PMID- 15891420 TI - Update on uveomeningoencephalitides. AB - PURPOSE OF REVIEW: Uveomeningoencephalitides, commonly known as Vogt-Koyanagi Harada (VKH) disease, are characterized by chronic bilateral granulomatous panuveitis involving the central nervous, auditory and integumentary systems. Visual prognosis is generally favourable, but outcomes in patients with VKH disease may vary. Also, the treatment of choice differs in different parts of the world. This review addresses the literature on the possible pathogenesis, diagnosis and treatment of this disorder. RECENT FINDINGS: Atypical presentations of VKH disease, as well as those associated with interferon-alpha therapy, have been reported. Most reports suggest an association with autoimmunity. The diagnostic criteria were revised by the International Workshop on VKH in 1999, allowing for the presence of different ocular findings in the early and late stages of the disease. New techniques have also been developed to aid in the rapid diagnosis of VKH disease and evaluation of treatment. Different routes of administration of corticosteroid and adjuvant therapy were tried, with positive results. SUMMARY: Although the pathogenesis of VKH disease is uncertain and antigen-specific treatment strategies have not yet been developed, reports increasingly suggest an autoimmune nature for uveomeningoencephalitides. Currently, systemic corticosteroid therapy remains the standard initial treatment. Different routes of administration are used to reduce the frequency of side effects of systemic corticosteroids, and there are various adjuvant therapies. With the aid of modern equipment, early diagnosis and prompt and appropriate treatment, resulting in better visual outcomes, can be anticipated. A large-scale, multinational, prospective study is warranted to determine the optimal initial therapy. PMID- 15891421 TI - Update on paraneoplastic syndromes. AB - PURPOSE OF REVIEW: This review discusses the varied clinical spectrum of neurologic paraneoplastic disorders, describes recent advances in our understanding of autoimmunity in these disorders, and outlines a practical clinical approach to patient management. RECENT FINDINGS: Paraneoplastic disorders may affect any part of the central or peripheral nervous system. Although relatively uncommon, these disorders are a significant cause of severe and permanent neurologic disability. Syndromes such as limbic encephalitis or opsoclonus-myoclonus should always raise suspicion of a paraneoplastic condition, but any paraneoplastic syndrome can also occur in patients without a neoplasm. Most neurologic paraneoplastic disorders are thought to be caused by an autoimmune reaction directed against 'onconeural' antigens expressed by neurons and tumor cells. Some syndromes such as Lambert-Eaton myasthenic syndrome and neuromyotonia are clearly mediated by autoantibodies. Much less is known about the immunopathogenesis of syndromes that affect the central nervous system, although a growing body of evidence implicates cellular immune effectors in causing neuronal injury. Many patients have circulating antineuronal antibodies, which can be useful in identifying a neurologic disorder as paraneoplastic and in finding the associated neoplasm. Early diagnosis of the neurologic disorder and prompt initiation of tumor treatment probably increase the likelihood of neurologic improvement. SUMMARY: Neurologists must be able to recognize the clinical manifestations of neurologic paraneoplastic disorders, and to distinguish them from other causes of neurologic dysfunction in cancer patients. Early diagnosis of paraneoplastic syndromes maximizes the likelihood of a favorable oncologic and neurologic outcome. PMID- 15891422 TI - Bibliography. Current world literature. Demyelinating diseases. PMID- 15891424 TI - On communication, humanism, and the brief encounter. PMID- 15891425 TI - Novel monitoring techniques for use with procedural sedation. AB - PURPOSE OF REVIEW: A variety of pharmacologic agents used for procedural sedation in children to reduce pain and anxiety may produce respiratory depression and hypotension. Although standard monitoring guidelines include oxygen saturation, this measurement is limited as a guide to respiratory function. This review discusses two new monitoring techniques recently introduced to the pediatric emergency department that facilitate procedural sedation and reduce potential adverse effects of the medications administered. RECENT FINDINGS: Capnography via an end-tidal carbon dioxide monitor measures carbon dioxide concentrations during ventilation. This measurement is independent of oxygen saturation and thereby aids the clinician in identifying hypoventilation and apnea in the sedated patient at an earlier stage than conventional monitoring. The bispectral index monitor objectively measures the depth of sedation by analyzing electroencephalogram signals from a cutaneous probe. This tool enables the physician to titrate sedative medications to a desired effect and thereby reduce the risks associated with oversedation. SUMMARY: Studies have illustrated the use of both devices as adjuncts to current standard monitoring of children in the outpatient setting. These modalities will facilitate the efficacy of procedural sedation in children and improve safety by enabling early recognition of hypoventilation and by reducing the risk of oversedation in children undergoing procedural sedation. PMID- 15891426 TI - Controversies in rapid sequence intubation in children. AB - PURPOSE OF REVIEW: Rapid sequence intubation is the method of choice for intubation of the emergency department patient. The purpose of the present review is to address several controversies pertaining to emergency department rapid sequence intubation of children. RECENT FINDINGS: The topics covered in this review include the determination of the appropriate clinician to perform emergency department intubation, the use of atropine and lidocaine as premedications, the choice of sedative agents depending upon the clinical scenario, and the choice of neuromuscular blockade agent. Concerning these topics: The literature supports that emergency department physicians, with appropriate training, successfully perform intubation in most patients. Limited data exist to determine the appropriate use of atropine and lidocaine for rapid sequence intubation. Etomidate has clearly become a preferred sedative for rapid sequence intubation with a low risk of cardiovascular side effects. Thiopental and propofol may more readily provide adequate sedation as compared with etomidate but both have the potential to reduce blood pressure. Succinylcholine arguably remains the preferred neuromuscular blockade agent for rapid sequence intubation in most children. The side effects of succinylcholine occur in relatively predictable circumstances. Rocuronium is a commonly used nondepolarizing paralytic agent but its prolonged duration of action must be weighed against the risk of side effects associated with succinylcholine. SUMMARY: Though more research is needed, the available data allow for the development of protocols that will result in a rational, scenario-based approach to rapid sequence intubation in children. PMID- 15891427 TI - Rapid viral testing in the evaluation of the febrile infant and child. AB - PURPOSE OF REVIEW: This review focuses on rapid viral testing in the febrile infant and child. Recent literature is reviewed regarding physician decision making, antibiotic use, ancillary testing use, and rate of serious bacterial infections concurrent with viral disease. RECENT FINDINGS: Two recent studies detail the use of rapid testing of influenza. The impact on the use of ancillary testing and antibiotic prescribing practices based on the knowledge provided by rapid viral testing has been evaluated. Physician awareness of a rapid diagnosis of influenza significantly reduced the number of laboratory tests and radiographs ordered and their associated charges, decreased antibiotic use, increased antiviral use, and decreased length of time to discharge. The rate of serious bacterial infections coexisting with influenza illness has also been studied. Researchers concluded that the prevalence of serious bacterial infections is lower in febrile children with influenza A infection. Another two studies evaluated respiratory syncytial virus-positive febrile infants and their risk of serious bacterial infection. Both studies independently noted that febrile infants with respiratory syncytial virus infections are at significantly lower risk of serious bacterial infection than febrile infants without respiratory syncytial virus infection. The rate of urinary tract infections remained significant in febrile respiratory syncytial virus-positive infants, however. SUMMARY: Various studies have documented the impact of rapid viral testing in the evaluation and management of febrile infants and children. There is insufficient evidence to change current clinical practice algorithms for young febrile infants and children. Continued research will affect future guidelines and algorithms in the management of febrile infants and children. PMID- 15891428 TI - Renal replacement therapy in pediatric critical care medicine. AB - PURPOSE OF REVIEW: The use of continuous renal replacement therapy in pediatric critical care has increased in the last 10 years. Adaptive makeshift machinery has been replaced with dedicated volumetric and/or gravimetric devices that afford accurate blood flow and ultrafiltrate production rates. While renal dysfunction continues to be related to primary renal disease, the incidence of secondary causes of acute renal failure continue to grow, especially in patients following cardiothoracic surgery, bone marrow transplantation, respiratory failure and multi-organ dysfunction syndrome. RECENT FINDINGS: Although much of the outcome data for continuous renal replacement therapy has been retrospective in nature, these therapies are safe for use in the sickest of intensive care unit patients. Moreover, early data from the prospective pediatric continuous renal replacement therapy registry suggests that early intervention with continuous renal replacement therapy, as well as goal-directed fluid resuscitation may lead to improved survival in critically ill patients. In patients with sepsis and septic shock, continuous renal replacement therapy offers a means for blood purification. SUMMARY: Though randomized placebo controlled trials are lacking at this time, center-based results suggest that continuous renal replacement therapy may prove beneficial to critically ill patients with sepsis and/or septic shock. PMID- 15891429 TI - Idiopathic pulmonary arterial hypertension in children. AB - PURPOSE OF REVIEW: Until recently, the diagnosis of idiopathic pulmonary arterial hypertension was virtually a death sentence, particularly for children. Although there is no cure for idiopathic pulmonary arterial hypertension, recent medical advances have dramatically changed the course of this disease in children. A review of some of the latest medical advances will provide the reader with a better understanding of the most current treatment options for children with idiopathic pulmonary arterial hypertension. RECENT FINDINGS: The literature reviewed demonstrate sustained clinical and hemodynamic improvement in children with various types of pulmonary arterial hypertension as well as increased survival in patients with idiopathic pulmonary arterial hypertension using current treatment strategies. SUMMARY: This article will provide an overview of how the current diagnostic and treatment strategies of idiopathic pulmonary arterial hypertension in children have advanced over the last several years and how this impacts on clinical practice. PMID- 15891430 TI - Pulmonary edema in infants and children. AB - PURPOSE OF REVIEW: To provide an overview of the pathogenesis of pulmonary edema and describe recent discoveries related to the clearance of airspace fluid and potential new therapies for this life-threatening disorder. RECENT FINDINGS: It is clinically important to determine the mechanisms responsible for the clearance of fluid from the airspaces. At birth inadequate clearance of fetal lung liquid is one of the two mechanisms leading to respiratory distress syndrome in the premature infant. Adults with heart failure or adult respiratory distress syndrome survive when they had active absorption of airspace fluid, yet have greater morbidity or die when they show no evidence of active fluid clearance. Humans who are susceptible to high-altitude pulmonary edema have less ability to actively transport fluid across their respiratory epithelium. SUMMARY: New approaches to increase the active clearance of fluid from the airspaces, combined with further improvements in the intensive care and monitoring of patients with serious illnesses, will lead to improved care for patients with lung diseases characterized by increased lung water content. PMID- 15891431 TI - Novel, mechanism-based therapies for cystic fibrosis. AB - PURPOSE OF REVIEW: Cystic fibrosis results from disruption of the biosynthesis or function of the cystic fibrosis transmembrane conductance regulator. Cystic fibrosis transmembrane conductance regulator plays a critical role in the regulation of epithelial ion transport. Restoration of cystic fibrosis transmembrane conductance regulator function should improve the cystic fibrosis phenotype. RECENT FINDINGS: Recent investigations affording a better understanding of the mechanism of dysfunction of mutant cystic fibrosis transmembrane conductance regulators, as well as the roles of cystic fibrosis transmembrane conductance regulator in regulating epithelial ion transport, have led to development of therapeutic strategies based on repair or bypass of mutant cystic fibrosis transmembrane conductance regulator dysfunction. The former strategy, coined 'protein repair therapy,' is aimed at improving or restoring the function of mutant cystic fibrosis transmembrane conductance regulators, whereas the latter approach aims to augment epithelial ion transport to compensate for the absent function mutant cystic fibrosis transmembrane conductance regulator. SUMMARY: Strategies to improve mutant cystic fibrosis transmembrane conductance regulator function or to bypass mutant cystic fibrosis transmembrane conductance regulator function hold great promise for development of novel therapies aimed at correcting the underlying pathophysiology of cystic fibrosis. PMID- 15891433 TI - Outcomes from the correction of anorectal malformations. AB - PURPOSE OF REVIEW: Anorectal malformations have been recognized and managed since antiquity, with surgical treatment evolving to maximize anatomic reconstruction, avoid complications, and understand mechanisms of incontinence, ultimately leading to improved quality of life for patients. This review describes recent advances in the management of anorectal malformations, including prenatal diagnosis, newborn treatment, surgical correction, and postoperative care. RECENT FINDINGS: Surgical treatment has improved with better understanding and exposure of anatomy and appreciation of the intimate relation between rectum and urinary tract. Repair of cloacal malformations has evolved to include the total urogenital mobilization and an appreciation of the complex associated Mullerian anomalies. The importance of associated urologic, gynecologic, neurologic, and orthopedic malformations has been recognized. Addition of a bowel management program to patients' postoperative care has increased dramatically the number of children who are clean and dry. SUMMARY: Management of anorectal malformations requires an accurate clinical diagnosis, proper newborn treatment, meticulous anatomic reconstruction, and comprehensive postoperative care with the goal of having a child who is clean and dry, with an excellent quality of life, because they either have the capacity for continence or can be kept artificially clean with a comprehensive bowel management program. PMID- 15891434 TI - Outcomes from pediatric solid organ injury: role of standardized care guidelines. AB - PURPOSE OF REVIEW: Recent advances in the delivery of trauma and critical care in children have resulted in improved outcome following major injuries. It is imperative that physicians who treat injured children familiarize themselves with current treatment algorithms for abdominal trauma. Important contributions have been made in the diagnosis and treatment of children with abdominal injury by radiologists and endoscopists. RECENT FINDINGS: This report examines the impact of consensus guidelines in the treatment of blunt abdominal solid organ injuries. Consensus guidelines for treatment of children with isolated spleen and liver injury are reviewed demonstrating conformity of care and significant reduction of resource utilization without adverse sequelae. Review of large datasets indicate contrasting rates of splenectomy depending on the expertise of the institution, emphasizing the need for wide dissemination of guidelines. SUMMARY: Clinical experience and published reports addressing specific concerns about the nonoperative treatment of children with solid organ injuries and recent radiologic and endoscopic contributions have made pediatric trauma care increasingly nonoperative. Although the trend is in this direction, the pediatric surgeon should remain the physician-of-record in the multidisciplinary care of critically injured children. PMID- 15891435 TI - Does surgical subspecialty care come with a higher price? AB - PURPOSE OF REVIEW: Pediatric surgical specialization has been occurring over the past few decades to improve the outcomes of pediatric surgical care. There is sparse data to support the benefit to children from these educational efforts of these dedicated individuals. This literature review focuses on recent studies and the data documenting improvement in pediatric surgical care. RECENT FINDINGS: Studies from the field of pediatric urology, pediatric general surgery and pediatric neurosurgery are cited. Studies compared outcome data: within a children's hospital between pediatric urology specialists and general urologist performing the same operation; within a healthcare system between pediatric general surgeons and adult general surgeons for appendectomies; population based data for pyloromyotomies performed by general surgeons and pediatric general surgeons; and finally a database study of specialty neurosurgeons and hospital factors affecting outcomes. SUMMARY: Each study documented better pediatric patient surgical outcomes under the care of a pediatric subspecialty trained surgeon regardless of discipline. Those studies that studied costs showed more cost effective care was delivered by pediatric subspecialized surgeons. PMID- 15891436 TI - Endoscopic treatment of vesicoureteral reflux. AB - PURPOSE OF REVIEW: The treatment of vesicoureteral reflux in children has seen a shift from invasive surgery to endoscopic management in recent years. Early studies demonstrate favorable outcomes but do not reach the success of surgery. Subspecialization also may play a role in the successful outcomes of all reflux management. This review focuses on the recent literature regarding endoscopic treatment of reflux and the role of subspecialization in treatment of children. RECENT FINDINGS: Dextranomer/hyaluronic acid copolymer has emerged as the favored bulking agent since its Food and Drug Administration approval in the United States. Although success is defined somewhat differently than surgery, overall success rates range from 82-89% with initial injection of dextranomer/hyaluronic acid. The outpatient nature of the procedure and its high success rate have led to a shift in treatment strategies especially in Europe, where endoscopic management is often considered first-line therapy. Extravesical reimplants can be performed on an outpatient basis safely. Subspecialization leads to improved outcomes with no difference in cost. SUMMARY: Endoscopic treatment of reflux is promising, but long-term results are not available to ascertain the efficacy of treatment into adulthood. Open surgery remains the gold standard. PMID- 15891437 TI - The Aristotle method: a new concept to evaluate quality of care based on complexity. AB - PURPOSE OF REVIEW: Evaluation of quality of care is a duty of the modern medical practice. A reliable method of quality evaluation able to compare fairly institutions and inform a patient and his family of the potential risk of a procedure is clearly needed. It is now well recognized that any method that purports to evaluate quality of care should include a case mix/risk stratification method. No valuable method was available until recently in pediatric cardiac surgery. RECENT FINDINGS: The Aristotle method is a new concept of evaluation of quality of care in congenital heart surgery based on the complexity of the surgical procedures. Involving a panel of expert surgeons, the project started in 1999 and included 50 pediatric surgeons from 23 countries. The basic score adjusts the complexity of a given procedure and is calculated as the sum of potential for mortality, potential for morbidity and anticipated technical difficulty. The Comprehensive Score further adjusts the complexity according to the specific patient characteristics (anatomy, associated procedures, co morbidity, etc.). The Aristotle method is original as it introduces several new concepts: the calculated complexity is a constant for a given patient all over the world; complexity is an independent value and risk is a variable depending on the performance; and Performance = Complexity x Outcome. SUMMARY: The Aristotle score is a good vector of communication between patients, doctors and insurance companies and may stimulate the quality and the organization of heath care in our field and in others. PMID- 15891438 TI - Immunizations, neonatal jaundice, and animal-induced injuries. AB - PURPOSE OF REVIEW: To concisely review data published over the past year on three topics pertinent to the practicing pediatrician: immunizations, neonatal jaundice, and animal-induced injuries. RECENT FINDINGS: Updates on immunizations, including varicella and pneumococcus in the post-vaccine era, use of a polyvalent conjugated meningococcal vaccine, and influenza vaccination during a vaccine shortage are discussed. Mortality and morbidity from varicella, and risk of invasive disease from pneumococcal infections have declined. The Advisory Committee on Immunization Practices of the Centers for Disease Control and Prevention released guidelines for the use of a tetravalent meningococcal conjugate vaccine in adolescents 11 years and older. Infants at younger gestational age are at higher risk for developing hyperbilirubinemia. The American Academy of Pediatrics has released new guidelines emphasizing breastfeeding, systematic pre-discharge assessment for risk factors, early follow up and intervention. Use of home nursing services in the postnatal period is cost effective. Although the incidence of rabies is low, many receive postexposure prophylaxis. Spider bites cause fewer systemic effects in children than adults. Pet reptile carriage of salmonella necessitates handwashing after handling of animals. SUMMARY: Recommendations for adolescent meningococcal vaccination, and the impact of varicella and pneumococcal immunization on our communities reflect an evolution in our management of infectious diseases. The latest practice guidelines on hyperbilirubinemia emphasize close follow-up of all newborns after postpartum discharge and even more careful evaluation of those infants near term. Education on rabies prophylaxis, spider bites, and salmonella from pet reptiles should focus on judicious use of resources and the proper handling of pets. PMID- 15891439 TI - Bibliography. Current world literature. Emergency and critical care medicine. PMID- 15891440 TI - Why? PMID- 15891441 TI - Outcomes of chlorine exposure: a 5-year poison center experience in 598 patients. AB - BACKGROUND: The outcome after chlorine exposures has been poorly described. METHODS: We conducted a 5-year retrospective poison control center review of chlorine exposure cases. The inclusion criteria were self (or surrogate) reported concentrated 'chlorine' liquid (> or =10% concentration of sodium hypochorite) or tablet (> or =90% concentration) exposure. Two reviewers blinded to the main purpose of the study reviewed charts and a third reviewed 10% of the charts, and a kappa score was calculated. Parameters reviewed included the type of exposure (tablets, liquid or both), symptoms (cough, chest pain, etc.), time to symptom onset, treatment received, hospital referral rate and outcomes. RESULTS: A total of 598 patient records were reviewed, with a mean age of 29.63 years (range 11 82); 41.5% (248) were exposures to tablets and 53.5% (320) were exposed to liquid chlorine, with the remainder exposed to both. Complaints included shortness of breath, eye irritation, nasal complaints, cough, and skin complaints. Sixty-nine patients were evaluated at a healthcare facility (11 referred by a poison control center and 58 were self-referrals). Five patients were admitted and four out of the five had a history of reactive airway disease. The mean time to peak symptoms was 94.4 min, 63.2% reached peak symptoms within 30 min, and 82% within 120 min, with 74% of all patients asymptomatic within 150 min. All five admitted patients were discharged within 48 h, and all others remained clinically well at a 24 h telephone follow-up. CONCLUSION: Exposure to chlorine tablets and liquid rarely requires hospital referral with almost all symptoms resolving within 24 h. PMID- 15891442 TI - Antioxidant enzymes, occupational stress and burnout in workers of a prehospitalary emergency service. AB - BACKGROUND: Reactive oxygen species (ROS) play an important role in many diseases and aging. Scavenging enzymes represent one of several natural defence mechanisms against ROS-induced damage. Superoxide dismutase (SOD) and catalase are enzymes that are involved in antioxidant defence. Burnout is a syndrome of emotional exhaustion, depersonalization, and reduced personal accomplishment. OBJECTIVES: To determine the activity of two antioxidant enzymes, SOD and catalase, and their variation according to demographic and occupational variables in workers of a prehospitalary emergency service, and to analyse the relationship between antioxidant enzyme activities and burnout 111 healthy workers of a prehospitalary emergency service were surveyed. METHODS: SOD activity was analysed by the Minami and Yoshikawa method. Catalase activity was measured by the method of Aebi. To measure burnout, the Maslach Burnout Inventory was used. RESULTS: No significant variations with respect to sex were detected in SOD and catalase activities and levels of burnout. Significant differences in SOD activity between the control group and workers of a prehospitalary emergency service (P<0.001) were found. SOD activity was higher in workers on night and evening shifts (P<0.001). These workers also obtained higher scores in burnout subscales, but without statistical significance. PMID- 15891443 TI - Metoclopramide induced dystonia in children: two case reports. AB - Metoclopramide is a dopamine antagonist that is widely used in gastroesophageal disease and chemotherapy-induced emesis in the paediatric population. It is also prescribed in nausea and vomiting caused by respiratory tract infections and enteritis in practice. The primary side-effect of the drug is extrapyramidal reactions with incidences as high as 25% in children. We report two cases, one of which was referred to our emergency department as encephalitis and the other as tetany, but which were just acute dystonic reactions caused by metaclopramide, even though the patients had used the drug in the recommended dosages. The adverse effects of the drug can be seen at normal doses. These dystonic reactions caused by metaclopramide can easily be confused with other diseases, because dystonia is not seen frequently in paediatric practice whatever the cause. PMID- 15891444 TI - Rupture of axillary artery and neuropraxis as complications of recurrent traumatic shoulder dislocation: case report. AB - Gleno-humeral dislocations are frequent, but associated complete transection of the axillary artery has only been reported sporadically. We present a case of complete rupture of the axillary artery associated with recurrent anterior dislocation of the shoulder, followed by a discussion of the likely mechanisms and the key learning points. PMID- 15891445 TI - Slow to make a speedy diagnosis. PMID- 15891446 TI - Predictors of an intermediate ventilation/perfusion lung scan in patients with suspected acute pulmonary embolism. AB - BACKGROUND: An intermediate ventilation/perfusion (V/Q) lung scan lacks diagnostic utility in excluding acute pulmonary embolism (PE). OBJECTIVE: To identify the potential predictors of an intermediate V/Q lung scan in patients with suspected acute PE. METHODS: A review of the medical records of 306 consecutive patients with suspected acute PE, in whom a V/Q lung scan was performed (interpretation using modified Prospective Investigation of Pulmonary Embolism Diagnosis criteria). RESULTS: A total of 274 patients (89.5%) had a non intermediate V/Q lung scan: 88 (28.8%) high probability, 149 (48.7%) low probability, and 37 (12.1%) near normal/normal. Thirty-two patients (10.5%) had an intermediate V/Q lung scan. Patients with an intermediate V/Q lung scan presented a significantly greater frequency of age greater than 70 years (81.2 versus 50.4%, P=0.01), previous cardiopulmonary disease (75 versus 46.3%, P=0.003), chronic obstructive pulmonary disease (34.4 versus 15.7%, P=0.01), and chest radiograph evidence of emphysema (18.7 versus 4.7%, P=0.008). We found no other significant differences in the rest of the characteristics studied between patients with an intermediate and a non-intermediate V/Q lung scan. CONCLUSION: The possibility of an intermediate V/Q lung scan is higher in elderly patients and in patients with previous cardiopulmonary disease (especially with chronic obstructive pulmonary disease). Emphysema is the only chest radiograph abnormality associated with a greater possibility of an intermediate V/Q lung scan. PMID- 15891447 TI - Traumatic aortic transection. AB - Traumatic aortic transection is an uncommon but often fatal injury. It is typically a high energy injury and may occur in the multiply injured patient. This injury is often missed. There is evidence that airbags and seat belts protect against these injuries. We present the case of a patient who survived. PMID- 15891448 TI - A teenager with angiographically normal epicardial coronary arteries and acute myocardial infarction after butane inhalation. AB - The most common cause of death among volatile substance abusers is sudden cardiac death. To the best of our knowledge, butane-induced myocardial infarction is a rare presentation among teenagers. We report on a 14-year-old male student who sustained cardiopulmonary arrest after sniffing seven canisters of butane. He was found to have extensive anterior myocardial infarction complicated by ventricular fibrillation. Cardiac catheterization revealed patent coronary arteries with severe anterolateral and apical left ventricular wall hypokinesia. We assume that cardiac complications after butane inhalations may partly be secondary to intense coronary artery spasm on the basis of the clinical and laboratory findings. This assumption may have major therapeutic implications in this patient population. PMID- 15891449 TI - Lisfranc fracture--dislocation: an easily overlooked injury in the emergency department. AB - Lisfranc fracture-dislocations are rare, and their correct diagnosis is important. Misdiagnosis is common and can lead to chronic disability. Two cases of Lisfranc fracture-dislocations are presented and emphasis is made on the diagnosis in the Emergency Department. PMID- 15891451 TI - Interaction of ionizing radiation with matter. AB - The basic physical mechanisms by which different kinds of ionizing radiation interact with matter are described. Understanding of these processes, coupled with detailed knowledge of atomic structure, provides much of the foundation for the theory and practice of radiological health protection today. PMID- 15891452 TI - Radiation biology: concepts for radiation protection. AB - The opportunity to write a historical review of the field of radiation biology allows for the viewing of the development and maturity of a field of study, thereby being able to provide the appropriate context for the earlier years of research and its findings. The pioneering work of Muller, Sax, and McClintock, and many others, has stood the test of time. The idea that x-rays could damage the genetic material and result in interactions that could lead to gene mutations and a range of chromosomal alterations is now interpretable in terms of induced DNA damage and errors of DNA repair. The expanded idea that such genetic alterations can be induced by DNA damage that is produced by one or two tracks of ionizing radiation remains the mainstay of radiation biology. The impact of the more recent molecular approaches to unraveling the mechanism behind this simple concept has confirmed this fundamental observation. The remarkable advances have allowed for a fairly complete understanding of the specific types of DNA damage induced by ionizing radiations and the pivotal role played by the errors of repair of double-strand breaks. Given our considerably enhanced knowledge of the details of the DNA repair processes involved, misrepair is a very unlikely event. The role of potential confounders of the concept of dose-response (e.g., bystander effects, genomic instability, and adaptive responses) is taking on a growing importance to the field. The evolving need is to begin to consider mechanistically-based dose-response models for cancer risk such that any potential impact of confounders on the response at low, environmental doses can be assessed. Thus, radiation biology research has always had a focus on how best to protect human health from radiation exposures and will continue to do so. PMID- 15891453 TI - External dosimetry and personnel monitoring. AB - The early history of our profession, as well as the discoveries that form its foundations, provide an interesting background on which to assess our progress in measuring exposure to ionizing radiation (i.e., dosimetry). One of the first challenges faced by scientists working with radiation and radioactivity was appropriate measurement techniques. has provided an excellent review of the history and development of radiation detection techniques and devices. In the early 20th century, some of these techniques were extended to measurements of the doses received by the individual scientists although it is not known exactly when organized personnel monitoring actually began. This manuscript will discuss some of the history of external dosimetry (specifically personnel monitoring), consider the developments that led to improved monitoring, attempt to assess the current status of the field and, also, to try to predict the future. The focus of this review will be primarily on personnel monitoring or personnel dosimetry. Obviously, it will not be possible to discuss every development in the field in detail and some choices have been made as to what to include and what to ignore. The author takes full responsibility for these choices. However, the goal is to provide sufficient coverage that the current situation and the future of personnel monitoring can be understood in the context of past developments. The discussion of the current status of external dosimetry will include some recent developments that hold great promise. In addition, a concern will be raised regarding the interpretation of the current U.S. federal regulations and the impact of the regulations on personnel dosimetry. Finally, there will be a discussion of what the future may hold in terms of the types of dosimeters and the approaches to personnel monitoring, in particular. PMID- 15891454 TI - Internal dosimetry: a review. AB - The field history and current status of internal dosimetry is reviewed in this article. Elements of the field that are reviewed include standards and models, derivation of dose coefficients and intake retention fractions, bioassay measurements, and intake and dose calculations. In addition, guidance is developed and provided as to the necessity of internal dosimetry for a particular facility or operation and methodology for implementing a program. A discussion of the purposes of internal dosimetry is included as well as recommendations for future development and direction. PMID- 15891455 TI - Recent history of the shielding of medical x-ray imaging facilities. AB - Since the discovery of x rays by Roentgen in 1895, a progression of reports containing recommendations to reduce the hazards from this "invisible light" have been published. These reports, generated by a number of expert national and international bodies, issued ever-evolving protection guidelines for shielding radiation workers and members of the public from the potential health effects of this radiation. In recent years, significant changes have been made in the recommended dose limits, and a number of articles that provide updated shielding data, parameters, and methodologies for x-ray imaging facilities have been published. Scientific Committee 9 (SC9) of the National Council on Radiation Protection and Measurements (NCRP) (formed with the support of the American Association of Physicists in Medicine), was given the charge of reviewing the literature and updating the NCRP shielding recommendations for x rays used for medical imaging purposes (i.e., for diagnostic and interventional procedures). This article focuses on the new data and models presented in these recent publications and on the recommendations of the current NCRP report, produced by SC9, for the design of radiation barriers in medical imaging facilities. PMID- 15891456 TI - Radiation shielding technology. AB - An historical review of the development of shielding techniques for indirectly ionizing radiation is presented, along with a summary of techniques at various levels of sophistication for shielding design and analysis. PMID- 15891457 TI - A history of radiation detection instrumentation. AB - A review is presented of the history of radiation detection instrumentation. Specific radiation detection systems that are discussed include the human senses, photography, calorimetry, color dosimetry, ion chambers, electrometers, electroscopes, proportional counters, Geiger Mueller counters, scalers and rate meters, barium platinocyanide, scintillation counters, semiconductor detectors, radiophotoluminescent dosimeters, thermoluminescent dosimeters, optically stimulated luminescent dosimeters, direct ion storage, electrets, cloud chambers, bubble chambers, and bubble dosimeters. Given the broad scope of this review, the coverage is limited to a few key events in the development of a given detection system and some relevant operating principles. The occasional anecdote is included for interest. PMID- 15891458 TI - Operational health physics. AB - A review of the operational health physics papers published in Health Physics and Operational Radiation Safety over the past fifteen years indicated seventeen general categories or areas into which the topics could be readily separated. These areas include academic research programs, use of computers in operational health physics, decontamination and decommissioning, dosimetry, emergency response, environmental health physics, industrial operations, medical health physics, new procedure development, non-ionizing radiation, radiation measurements, radioactive waste disposal, radon measurement and control, risk communication, shielding evaluation and specification, staffing levels for health physics programs, and unwanted or orphan sources. That is not to say that there are no operational papers dealing with specific areas of health physics, such as power reactor health physics, accelerator health physics, or governmental health physics. On the contrary, there have been a number of excellent operational papers from individuals in these specialty areas and they are included in the broader topics listed above. A listing and review of all the operational papers that have been published is beyond the scope of this discussion. However, a sampling of the excellent operational papers that have appeared in Health Physics and Operational Radiation Safety is presented to give the reader the flavor of the wide variety of concerns to the operational health physicist and the current areas of interest where procedures are being refined and solutions to problems are being developed. PMID- 15891459 TI - Medical health physics: a review. AB - Medical health physics is the profession dedicated to the protection of healthcare providers, members of the public, and patients from unwarranted radiation exposure. Medical health physicists must be knowledgeable in the principles of health physics and in the applications of radiation in medicine. Advances in medical health physics require the definition of problems, testing of hypotheses, and gathering of evidence to defend changes in health physics practice and to assist medical practitioners in making changes in their practices as appropriate. Advances in radiation medicine have resulted in new modalities and procedures, some of which have significant potential to cause serious harm. Examples included in this review include radiologic procedures that require very long fluoroscopy times, radiolabeled monoclonal antibodies, and intravascular brachytherapy. This review summarizes evidence that supports changes in consensus recommendations, regulations, and health physics practices associated with recent advances in radiology, nuclear medicine, and radiation oncology. Medical health physicists must continue to gather evidence to support intelligent but practical methods for protection of personnel, the public, and patients as modalities and applications evolve in the practice of medicine. PMID- 15891460 TI - Light water reactor health physics. AB - In this article an overview of the historical development of light water reactor health physics programs is presented. Operational health physics programs have developed and matured as experience in operating and maintaining light water reactors has been gained. Initial programs grew quickly in both size and complexity with the number and size of nuclear units under construction and in operation. Operational health physics programs evolved to face various challenges confronted by the nuclear industry, increasing the effectiveness of radiological safety measures. Industry improvements in radiological safety performance have resulted in significant decreases in annual collective exposures from a high value of 790 person-rem in 1980 to 117 person-rem per reactor in 2002. Though significant gains have been made, the continued viability of the nuclear power industry is confronted with an aging workforce, as well as the challenges posed by deregulation and the need to maintain operational excellence. PMID- 15891461 TI - Environmental health physics: 50 years of progress. AB - Environmental health physics is an interdisciplinary field, involving study of the release, transport, and fate of radioactive material in the environment. Further, it addresses the interaction of humans with radioactive materials within the ambient (outdoor) environment and with the environments associated with modern technology and lifestyles. It also involves both naturally occurring and artificially produced radionuclides with the former generally being by far the highest source of exposure. In fact, doses from naturally occurring radionuclides are increasingly being used as a benchmark for the establishment of dose rate limits for people. Because of the pioneering work of early environmental health physicists, models exist today that can be used to assess the potential impacts of new nuclear facilities prior to their operation. In fact, these people represent the branch of the health physics profession who conducted environmental monitoring programs and performed the associated research studies that led to the identification of the principal radionuclides of interest, the major pathways and mechanisms through which they expose people, and the doses that may result from radioactive materials in the natural and technologically enhanced environments. One of their most important contributions was the identification and quantification of many of the key parameters that serve as input to such models. Monitoring of nuclear weapons development facilities used during and after World War II was the initial stimulus for the establishment of environmental health physics programs. Thereafter, these programs were expanded both nationally and globally, as a result of the atmospheric weapons testing programs of nations such as France, the People's Republic of China, the former Soviet Union, the United Kingdom, and the United States. Additional stimuli were provided by the development of the commercial nuclear power industry. Current environmental programs, particularly within the U.S., focus on decontamination and decommissioning of dormant facilities from these earlier defense and commercial programs. The range of the environmental health physics aspects of these activities is the subject of this paper. Presented at the end of the paper is a summary of some of the more important lessons that have been learned. As will be noted, this is an exciting field that will present challenges to health physicists for years to come. PMID- 15891462 TI - A review of the history of U.S. radiation protection regulations, recommendations, and standards. AB - Shortly after the discovery of x rays by Wilhelm Konrad Roentgen in 1895, and the isolation of the element radium by Pierre and Marie Curie three years later, the fascination with and potential for an array of uses of ionizing radiation in medicine, science, and technology was born. As with any new technology, there was a need to balance both the beneficial and potential detrimental effects of uses of these new technologies for the advancement of humankind. In the early days, radiation hazards were not well understood. Over the decades increasing concerns in the scientific community and lay population demanded that standardized guidance and recommendations be developed for the use of ionizing radiation. Today, U.S. radiation protection standards and recommendations to protect the occupational worker, members of the general public, and the environment are numerous and complex. This review summarizes the history of the development and application of radiation protection standards and regulations to assure the safe use of radiation and radioactive materials. The evolution and roles of international and national scientific recommending and regulatory organizations that shape U.S. radiation protection policy are described and discussed. PMID- 15891463 TI - A history of the international commission on radiological protection. AB - Within twelve months of the discovery of x rays, papers appeared in the literature reporting adverse effects from high exposure. By the time of the First World War, several countries were proposing restrictions for the exposure of radiation workers. In 1925, the first International Congress of Radiology, held in London, considered the need for a protection committee, which it established at its second Congress in Stockholm in 1928. This paper traces the history of the development, by ICRP, of its policies and the personalities involved in their development from its inception up to the modern era. The paper follows the progress from the early controls on worker doses to avoid deterministic effects, through the identification of stochastic effects to the concerns about increasing public exposure. The key features of the Recommendations made by ICRP from 1928 up to the current 1990 version are identified. PMID- 15891464 TI - The Health Physics Society: a 50-year chronology. AB - In 2005, the Health Physics Society celebrates its 50th anniversary. During its half century of existence, the Society has evolved from a small group of operationally oriented radiation protection scientists into an organization of several thousand with a recognized and respected peer reviewed journal. The bold decision to form an organization devoted to radiation safety in the United States spurred the formation of such societies in other countries and led to the formation of an international confederation of national societies, which has facilitated scientific communication and exchange worldwide. It also led to the formation of a separate body for the certification of health physicists, which in turn has grown into an academy with more than 1,000 members, and was instrumental in providing basic information establishing the need for a registry of radiation protection technologists. PMID- 15891470 TI - How to navigate and sail the waters around OASIS M0440. PMID- 15891471 TI - Pay for performance--Medicare's new payment model? PMID- 15891472 TI - A home visitation program welcomes home first-time moms and their infants. AB - This article describes a creative maternal and infant home visiting program for first-time parents. Two-year outcomes indicate the program improved parents' infant safety knowledge, positively affected the mother's decision to breastfeed, and promoted infant primary care visits in a cost-effective way. PMID- 15891473 TI - Disease management: a new and exciting opportunity in home healthcare. AB - Disease management programs are beginning to encompass providers across the healthcare continuum, including home healthcare. The premise behind disease management is that coordinated, evidence-based interventions can be applied to the care of patients with specific high-cost, high-volume chronic conditions, resulting in improved clinical outcomes and lower overall costs. Outcomes data (actual results) are central in this approach to patient care. PMID- 15891475 TI - Creative inservices to meet education requirements. PMID- 15891476 TI - Wound dressings: a myriad of challenging decisions. AB - Choosing the appropriate wound dressing is critical to positive patient outcomes. This article helps the nurse choose dressings based on wound care principles and the comprehensive assessment. Tools are provided to choose the different products that can be used as the wound progresses through the stages of healing. PMID- 15891477 TI - Care at the time of death. PMID- 15891479 TI - The clinician's role in Medicare prospective payment: part 4--Medicare adjustments. PMID- 15891480 TI - The power of one. PMID- 15891481 TI - Integration of technology and facility design: implications for nursing administration. PMID- 15891482 TI - Generating enthusiasm for nursing research. PMID- 15891483 TI - Managing patient care error: nurse leaders' perspectives. PMID- 15891484 TI - When the culture of disability meets ethnicity: a challenge for providing culturally competent care. PMID- 15891485 TI - Evaluating the effectiveness of a certificate in clinical leadership program on the performance of nurse managers. PMID- 15891486 TI - Nurse executive transformational leadership and organizational commitment. AB - OBJECTIVE: To investigate the relationship between nurse executive leadership and organizational commitment among nurses in acute care hospitals. BACKGROUND: A key challenge for organizations is to maximize the contributions of all workers by cultivating their commitment. Nurse leaders are in a position to influence organizational commitment among nurses. METHODS: The theoretical constructs underlying this study are the transformational leadership theory and the Etzioni's organizational theory. A cross-sectional, field survey of nurse executives, nurse managers, and staff nurses was conducted to assess nurse executive transformational and transactional leadership and their relationship to organizational commitment. Hypotheses were tested using correlational analysis, and univariate statistics were used to describe the sample. RESULTS: An inverse relationship between nurse executive transformational and transactional leadership and alienative (highly negative) organizational commitment was statistically significant. A positive association was demonstrated between nurse executive leadership and nurse manager leadership. CONCLUSIONS: This study supports the effect of nurse executive leadership on nurse manager leadership and on organizational commitment among nurses despite role distance. To the extent that transformational leadership is present, alienative organizational commitment is reduced. This relationship shows the importance of nurse executive leadership in organizational involvement among nurses in the dynamic context of contemporary hospital settings. PMID- 15891487 TI - Engaging nurse leaders in health services research. AB - OBJECTIVE: To describe effective methods to engage nurse leaders in structured interview research. BACKGROUND: The American Organization of Nurse Executives has identified the support of nurse leader participation in research as a key strategy to achieving its education and research objectives, particularly with regard to identifying and documenting nursing sensitive outcomes. Previous studies have delineated several methods to increase participation. METHODS: Nurse leaders of California labor and delivery units reporting more than 50 deliveries during 2002 were asked to participate in a structured interview about staffing and clinical policies on their unit. Recruitment methodology involved 3 levels of intervention, including introductory letters, follow-up contacts, and personal encouragement from senior regional leaders and nurse executives. RESULTS: Of the 268 eligible hospitals, 225 (84%) participated in the study. Fifty-four percent (n = 114) of the structured interviews were scheduled upon initial and second contacts by the research coordinator, and 73% (n = 161) were completed at the first scheduled appointment. Unit managers or directors personally completed 91% (n = 205) of the interviews, with only 20 (8.9%) designated to a staff nurse. CONCLUSIONS: Nurse leader participation is essential to the success of research dependent on collecting information regarding nurse practices and clinical processes. Using a multilevel approach to engage nurse leaders in research, such as endorsements and "detailing" by opinion leaders, phone calls, electronic mail, and incentives, is an effective strategy. PMID- 15891488 TI - A review of instruments measuring nurse-physician collaboration. AB - OBJECTIVE: To review instruments used to measure nurse-physician collaboration and compare the strengths and potential opportunities of each instrument. BACKGROUND: Nurse-physician collaboration has been studied using a variety of instruments. The ability to generalize the outcomes of studies and build on the findings is predicated on acceptable validity and reliability metrics of these instruments. METHODS: A literature search using PubMed and Health and Psychological Instruments databases was conducted for articles published between 1990 and May 2004 to identify instruments measuring staff nurse-physician collaboration. After the instruments were identified, a second search was conducted to identify at least one peer-reviewed article describing the psychometrics of the instrument. Articles identified were then entered into the ISI Web of Science Citation Index to identify the instruments that had been used in at least 2 other studies. These selected instruments were then reviewed for the following information: background for the development of the tool, description of the tool, initial psychometric testing, and strengths and potential applications for each instrument. RESULTS: Five instruments met study criteria: the Collaborative Practice Scale, Collaboration and Satisfaction About Care Decisions, ICU Nurse-Physician Questionnaire, Nurses Opinion Questionnaire, and the Jefferson Scale of Attitudes Toward Physician Nurse Collaboration. CONCLUSIONS: The identified instruments have undergone initial reliability and validity testing and are recommended for future research on nurse-physician collaboration. PMID- 15891489 TI - Health, work variables, and job satisfaction among nurses. AB - BACKGROUND: Evidence from several studies suggests that there is widespread job dissatisfaction among nurses. Coupled with a critical shortage of RNs, this situation threatens the provision of safe healthcare. OBJECTIVE: The purpose of this study was to explore the relationships and relative contributions of selected work (stress, work load, weekends off), shift worker health (sleep, depression), and demographic variables (age, number of individuals needing care after work) to job satisfaction in a random, nationwide sample of 247 critical care RNs. METHODS: The Dillman Tailored Design Method of survey research was used to recruit participants and collect data. A descriptive, correlational design evaluated the relationships between the variables. RESULTS: There were no significant differences in these variables among self-defined day-, night-, and rotating-shift nurses. Hierarchical regression analyses indicated that more weekends off per month and less depression and emotional stress contributed significantly to job satisfaction in nurses. CONCLUSIONS: Improvements in scheduling and interventions designed to reduce depression and emotional stress may help to improve job satisfaction in nurses and aid in nurse recruitment and retention. PMID- 15891490 TI - Faces of the nursing shortage: influences on staff nurses' intentions to leave their positions or nursing. AB - OBJECTIVE: The objective of this study was to examine the relationship between organizational commitment, job satisfaction, and nurses' intention to leave their current position or nursing. BACKGROUND: Because retention of nurses is a critical issue for acute care institutions, study of the predictors of nurses' intention to leave (position or nursing) is essential. Of interest in predicting intention to leave are both organizational commitment and job satisfaction. METHODS: A mailed survey was conducted in which 787 respondents completed measures of satisfaction, organizational commitment, and personal and institutional demographics. All instruments had reliability estimates ranging from alpha = .76 to .92. RESULTS: Predictive relationships were found between financial status, organizational commitment, job and professional satisfaction, and intent to leave their current position (R = 0.42) and between financial status, professional satisfaction, and intention to leave nursing (R = 0.45) CONCLUSIONS: Organizational commitment has implications for nurses' intention to leave their current position but not nursing. Aspects of work and professional satisfaction predict both the intention to leave current position and nursing. Retention programs need to be focused on both organizational commitment and work and professional satisfaction. PMID- 15891491 TI - Promoting nurses' self-efficacy: a leadership strategy to improve practice. AB - OBJECTIVE: The purpose of this study was to investigate the interaction between structural empowerment, nursing leadership, and self-efficacy for nursing practice, to determine if self-efficacy could contribute to more professional nursing behaviors. BACKGROUND: Increasing self-efficacy for nursing practice may improve nurses' professional practice behaviors because individuals not only react to environmental influences, but also have the ability to exercise self influence to shape their social systems. A new theory was developed that suggests the interaction between structural empowerment, nursing leadership, and self efficacy may determine whether a nurse's practice behavior is either professional or more task-focused. METHODS: The study used a nonexperimental survey design. Path analysis was used to demonstrate both direct and indirect relationships. Mediation was also tested through Sobel's tests. RESULTS: Self-efficacy partially mediated the relationship between structural empowerment and professional practice behaviors. Sobel's tests were significant when self-efficacy mediated the relationship between structural empowerment and professional practice behaviors in the group who perceived strong nursing leadership, as well as in the model where nursing leadership was not included. CONCLUSIONS: Nurse managers may want to provide opportunities for enhancing self-efficacy, such as role modeling and verbal persuasion, recognizing that through improved self-efficacy, practice behaviors may improve. PMID- 15891492 TI - Media violence exposure and frontal lobe activation measured by functional magnetic resonance imaging in aggressive and nonaggressive adolescents. AB - OBJECTIVE: To understand better the relation between media violence exposure, brain functioning, and trait aggression, this study investigated the association between media violence exposure and brain activation as measured by functional magnetic resonance imaging (fMRI) in groups of normal adolescents and adolescents with disruptive behavior disorder (DBD) with aggressive features. METHODS: Seventy-one participants underwent neuropsychologic evaluation and assessment of exposure to violent media. Subjects also were evaluated with fMRI while performing a counting Stroop (CS) task. RESULTS: Frontal lobe activation was reduced in aggressive subjects compared with control subjects. In addition, differences in frontal lobe activation were associated with differences in media violence exposure. Specifically, activation during performance of the CS in control subjects with high media violence exposure resembled that seen in DBD subjects. CONCLUSIONS: Our findings suggest that media violence exposure may be associated with alterations in brain functioning whether or not trait aggression is present. PMID- 15891493 TI - Radiation-induced brain metabolic changes in the acute and early delayed phase detected with quantitative proton magnetic resonance spectroscopy. AB - OBJECTIVE: Quantitative proton magnetic resonance spectroscopy (MRS) was performed before and after radiation therapy to estimate its usefulness for evaluating radiation-induced metabolic brain changes. METHODS: Twenty patients with multiple brain metastases not having received any previous brain radiation were selected for the study. The total radiation dose varied from 40 (20 fractions) to 50 (25 fractions) Gy, with an opposition technique. MRS was performed just before irradiation, during the acute phase (n = 20, 8.5 +/- 4.6 days) and in the early delayed phase (n = 15, 3.6 +/- 0.5 months) after radiation. The concentration of N-acetyl-L-aspartate (NAA), choline-containing substance (Cho), and creatine/phosphocreatine (Cr) was quantified. RESULTS: The concentration of NAA decreased (P = 0.05 versus before radiation), and the concentration of Cho increased (P = 0.006 versus before radiation) during the early delayed phase. The concentration of Cr was not changed before or after radiation. CONCLUSIONS: Radiation-induced changes in brain metabolism were well detected with quantitative MRS in the early delayed phase. Quantitative MRS is a novel tool for estimating radiation-induced neurotoxicity. PMID- 15891494 TI - Brain apparent diffusion coefficient evaluation in pediatric patients with neurofibromatosis type 1. AB - OBJECTIVE: The most frequent intracranial appearance in children with neurofibromatosis type 1 (NF1) is represented by the presence of hyperintense lesions on T2-weighted images, the so-called "unidentified bright objects" (UBOs). Di Paolo demonstrated that these lesions represent foci of myelin vacuolization with increased water content. The aim of this study was to investigate the isotropic apparent diffusion coefficient (ADC) values within the UBOs and normal-appearing brain and at the regressed UBO sites. METHODS: Fifteen consecutive children with NF1 underwent magnetic resonance diffusion evaluation of the brain. Fifteen healthy age- and sex-matched children constituted the control group. Apparent diffusion coefficient maps were obtained, and regions of interest were placed bilaterally in 8 different areas. Two cortical areas were evaluated using single-pixel analysis. Apparent diffusion coefficient values within the UBOs were calculated by using irregular regions of interest. Regressed UBO sites were investigated by using circular regions of interest. Apparent diffusion coefficient values within the different areas were compared using a t test. RESULTS: Compared with the controls, NF1 patients showed higher ADC values (P < 0.001) in all locations. In the NF1 group, the mean ADC value in the UBOs was higher than in other locations (P < 0.001). The mean ADC value within the regressed UBO sites was higher than in the normal-appearing locations (P < 0.001). CONCLUSIONS: The higher ADC values in children with NF1 suggest an increase in water content of the normal-appearing brain. The UBOs are the areas with the highest water content. The regressed UBOs sites show higher water content than the normal-appearing areas. PMID- 15891495 TI - Registration and fusion of CT and MRI of the temporal bone. AB - OBJECTIVE: To present and evaluate a registration method to fuse complementary information of CT and MRI of the temporal bone. METHODS: CT and MRI of the temporal bone of 26 patients were independently registered 4 times. A manual, iterative, intrinsic, rigid, and retrospective registration method was used. Mean CREm (consistency registration error) was calculated as a reproducibility measurement. RESULTS: CREm was 0.6 mm (95% CI = 0.52-0.68 mm). T-test revealed no difference between pathologic and normal cases (t[102] = -1.71; P = 0.09). Time needed: 13 minutes. In the registered and fused datasets, important bony surgical landmarks (eg, facial nerve canal, inner ear) could be assessed in 3 dimensions relatively to tumor tissue (eg, acoustic schwannoma). Fluid distribution within partially obliterated cochleae could be assigned to either scalae. CONCLUSION: An accurate, reproducible registration and fusion method that improves tumor surgery and cochlea implantation planning with only minor changes to the clinical workflow was presented and described. We suggest this method in selected cases. PMID- 15891496 TI - Pneumatization of the inferior turbinates: incidence and radiologic appearance. AB - OBJECTIVE: The objective of this work was to determine the incidence and radiologic appearance of pneumatized inferior turbinates (PITs). METHODS: Pneumatized inferior turbinates were investigated among 2500 consecutive cases with coronal paranasal sinus computed tomography (CT) scans that were performed for different reasons between 2000 and 2004. RESULTS: Ten new PIT cases diagnosed with CT were identified. Pneumatization was diagnosed in 2 patients aged 7 and 14 years. Therefore, this is the first time that PIT has been described in the pediatric age group. Complete or incomplete pneumatizations were unilateral in 7 cases and bilateral in 3 cases. In bilateral PIT cases, pneumatizations were asymmetric. In 2 cases, only bulbous parts of the inferior turbinates (ITs) were involved. Inferior turbinate expansion related to pneumatization was not observed in any case. In cases with maxillary sinus communication, axial CT sections revealed a defect on the medial wall of the maxillary sinus. CONCLUSIONS: The incidence of PIT was found to be 1 in 250 cases. It is suggested that most PIT cases may be a result of articulation defects between the maxillary process of the palatine bone and the maxillary bone. In such a situation, the maxillary process of the palatine bone and the nasal process of the maxillary bone, which form the medial wall of the maxillary sinus at the IT level, can attach to the IT separately. PMID- 15891497 TI - Use of cross-sectional imaging in predicting surgical location of parotid neoplasms. AB - OBJECTIVE: The purpose of this study was to determine the diagnostic accuracy of using the retromandibular vein as seen on cross-sectional imaging to help differentiate superficial lobe from deep lobe tumors. METHODS: Of the patients who had parotid neoplasms between January 1997 and July 2002, we were able to identify 44 patients with preoperative imaging studies that were available for evaluation. The films were reviewed by a single head and neck radiologist to determine whether the neoplasms involved the superficial, deep, or both lobes of the parotid gland (total). The lateral margin of the retromandibular vein was used as a marker for the facial nerve, since the nerve is not always visible on CT and MRI scans. The radiologist's findings were then compared with the findings during surgery. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of predicting the location of neoplasms were then calculated. RESULTS: For lesions in the superficial lobe, cross-sectional imaging was able to predict the location of the neoplasm with a sensitivity of 0.91 (95% CI, 0.70-0.98), specificity of 0.86 (95% CI, 0.63-0.96), PPV of 0.88 (95% CI, 0.67-0.97), and NPV of 0.90 (95% CI, 0.67-0.98). For lesions in both lobes (total), cross-sectional imaging was able to predict the location of the neoplasm with a sensitivity of 0.94 (95% CI, 0.68-0.99), specificity of 0.89 (95% CI, 0.71-0.97), PPV of 0.83 (95% CI, 0.58-0.96), and NPV of 0.96 (95% CI, 0.78 0.99). CONCLUSION: Use of the retromandibular vein as a marker for the facial nerve is a sensitive method for identifying the location of parotid gland neoplasms on cross-sectional imaging. This supports the accuracy of using preoperative imaging to detect the position of parotid neoplasms with respect to the facial nerve. PMID- 15891498 TI - Proton magnetic resonance spectroscopy and diffusion-weighted imaging of central nervous system whipple disease. AB - A 72-year-old man presented with a 6-month history of dysphagia, fatigue, 60-lb weight loss, and central nervous system (CNS) deficits. Diffusion-weighted magnetic resonance imaging (MRI) of the brain showed mildly elevated nonrestricted apparent diffusion coefficients in the middle cerebellar peduncles, and magnetic resonance spectroscopy (MRS) showed decreased N-acetylaspartate and creatine with increased choline. Diffusion-weighted MRI and MRS offer noninvasive methods to help evaluate in vivo physiologic changes of CNS involvement in Whipple disease. PMID- 15891499 TI - Tyrosinemia: computed tomography, magnetic resonance imaging, diffusion magnetic resonance imaging, and proton spectroscopy findings in the brain. AB - In a 5-month-old boy with tyrosinemia, computed tomography revealed diffuse hypodensity in the centrum semiovale. Magnetic resonance (MR) imaging revealed partial signal differences in the white matter and perirolandic regions, and the posterior limbs of the internal capsules revealed higher signal compared with the remainder of the white matter. There were high-signal changes (restricted water diffusion) in the corresponding regions on images of diffusion-weighted MR imaging (b = 1000 s/mm). This likely represented the presence of intramyelinic edema attributable to status spongiosus. Proton MR spectroscopy (repetition time = 1500 milliseconds, echo time = 40 milliseconds) from the lesion sites revealed 2 separate prominent peaks spread between 3.4 and 3.9 ppm. These peaks could represent the CH and CH2 aliphatic protons of the tyrosine molecule. PMID- 15891500 TI - Value of magnetic resonance imaging for muscle denervation syndromes of the shoulder girdle. AB - Clinical evaluation of neuromuscular disorders typically consists of obtaining a detailed clinical history, physical examination, and electrophysiologic examinations. Electrodiagnostic examinations significantly aid in distinguishing between myopathy, neuropathy, and neuromuscular disorders. Electrodiagnostic examinations also assist in determining the severity and extent of disease. Progress can also be monitored on follow-up testing. The benefit of magnetic resonance imaging (MRI) in neuromuscular disease evaluation lies primarily in identifying a specific underlying gross pathologic cause and its location in the neuraxis as well as in identifying associated secondary findings. In some cases, MRI is particularly helpful when a solitary, small, deep muscle is affected. Imaging can be useful in assessing clinical progress in some cases. Causes of muscle denervation include mass lesions and trauma as well as infectious, autoimmune, and idiopathic causes. This article illustrates the common denervation syndromes that involve the shoulder girdle: Parsonage-Turner syndrome, quadrilateral space syndrome, and suprascapular neuropathy. By demonstrating the exact muscles involved and spared, MRI noninvasively identifies the level of nerve insult in the neuraxis. Furthermore, in cases in which a mass is responsible for denervation, MRI can directly show the cause and aid in treatment planning. PMID- 15891501 TI - Lobulated schwannoma of the median nerve: pitfalls in diagnostic imaging. AB - The authors report an unusual case of volar wrist swelling with the appearance of a simple ganglion on magnetic resonance imaging (MRI); however, operative and histologic examination identified the swelling as an ancient schwannoma. Ancient schwannomas mostly present as a solitary tumor. A lobulated tumor in this location has not been described previously. The pathologic findings of these uncommon tumors and the difficulties encountered in accurate preoperative diagnosis are discussed. PMID- 15891502 TI - "Black" contrast effect during magnetic resonance arthrography attributable to inadvertent administration of excessive gadolinium chelates. AB - Excessive intra-articular administration of gadolinium chelates during magnetic resonance (MR) arthrography results in decreased signal throughout the joint on MR images. A case with this condition during MR arthrography of the hip is described, and a possible solution when this problem occurs (ie, performing the MR imaging examination after a waiting period of several hours) is suggested. PMID- 15891503 TI - Multidetector computed tomography of fracture-separation of the distal humeral epiphysis. AB - A case of fracture-separation of the distal humeral epiphysis that was diagnosed using unenhanced multidetector computed tomography is reported. Fracture separation of the distal humeral epiphysis is a rare injury of young children that is difficult to identify on routine radiographs and is frequently misdiagnosed at the time of initial evaluation. PMID- 15891504 TI - High-resolution computed tomography features of nonspecific interstitial pneumonia and usual interstitial pneumonia. AB - OBJECTIVE: To assess the accuracy of high-resolution computed tomography (HRCT) in the diagnosis of nonspecific interstitial pneumonia (NSIP). We hypothesized that the computed tomography (CT) features of NSIP could be distinguished from those of usual interstitial pneumonia (UIP). METHODS: The HRCT images of 47 patients with surgical lung biopsy-proven NSIP (n = 25) and UIP (n = 22) were independently reviewed by 2 thoracic radiologists. Predominant imaging patterns, most likely diagnosis, and diagnostic level of confidence were recorded. A confident HRCT diagnosis of NSIP was based on the presence of spatially uniform, bilateral, basal-predominant ground-glass and/or reticular opacities with little if any honeycombing, whereas UIP was confidently diagnosed if a spatially inhomogeneous, bilateral, peripheral, basal-predominant pattern of reticular opacities and honeycombing with little if any ground-glass attenuation was identified. RESULTS: A predominant pattern of ground-glass and/or reticular opacity with minimal to no honeycombing was demonstrated in 48 (96%) of 50 readings in patients with NSIP. Conversely, the presence of honeycombing as a predominant feature had a predictive value of 90% for UIP (P < 0.001). Usual interstitial pneumonia was more likely than NSIP to be subpleural and patchy (P < 0.001). A confident CT diagnosis of NSIP and UIP was correct in 73% and 88% of cases, respectively. The correctness of a CT diagnosis made at intermediate or high confidence was 68% and 88%, respectively. The kappa value for distinction of NSIP from UIP was 0.72. CONCLUSION: In contrast to previous reports, NSIP can be separated from UIP in most cases. The presence of honeycombing as a predominant imaging finding is highly specific for UIP and can be used to differentiate it from NSIP, particularly when the distribution is patchy and subpleural predominant. The presence of predominant ground-glass and reticular opacity is highly characteristic of NSIP, but there is a subset of patients with UIP who have this pattern and may require biopsy for differentiation from NSIP. PMID- 15891505 TI - Fluorodeoxyglucose positron emission tomography pattern of pulmonary lymphangitic carcinomatosis. AB - OBJECTIVE: The purpose of this study was to evaluate the F-fluorodeoxyglucose (FDG) positron emission tomography (PET) uptake in pulmonary lymphangitic carcinomatosis identified on computed tomography (CT) in patients with lung cancer. METHODS: This was a retrospective analysis of F-FDG PET images in 7 patients (4 male and 3 female, mean age: 56.6 +/- 6.6 years) with lung cancer with a CT-based diagnosis of lymphangitic carcinomatosis. The F-FDG PET scans in a group of 7 patients (4 male and 3 female, mean age: 42.1 +/- 5.66 years) with normal chest CT scans served as a control group. Mean standardized uptake values (SUVs) were calculated based on average tumor uptake, initial injected activity, and body weight. RESULTS: The intensity of F-FDG uptake in diseased lung is significantly greater than in corresponding normal contralateral lung or in the lungs of normal controls (P = 0.003). The ratio of the SUV of lung with lymphangitic carcinomatosis to corresponding contralateral normal lung was significantly increased (P = 0.006), and the ratio of the SUV of mediastinal blood pool to lung with lymphangitic carcinomatosis was significantly decreased (P = 0.0002). CONCLUSION: There is diffuse increased FDG uptake in the lung corresponding to the CT pattern of lymphangitic carcinomatosis. PMID- 15891506 TI - High-resolution CT in silicosis: correlation with radiographic findings and functional impairment. AB - OBJECTIVE: To assess high-resolution computed tomography (HRCT) findings in silicosis and to better define the role of HRCT in early detection of parenchymal abnormalities in silica-exposed workers. METHODS: Forty-one stone carvers were evaluated with chest radiographs (CR), HRCT, and pulmonary function tests (PFT). Inter-reader agreement was calculated using the kappa statistic (k). Correlation between radiographic and HRCT profusion scores and PFT was assessed using the Spearman correlation coefficient. RESULTS: The most common HRCT findings were branching centrilobular structures, seen in 28/41 workers (68.3%). Nodules consistent with silicosis were detected in 53.7% workers on CR and in 56.1% workers on HRCT. Inter-reader agreement for diagnosis of silicosis was better on HRCT (k = 0.84) than on CR (k = 0.54). Small opacity profusion on HRCT correlated inversely with total lung capacity and FVC%. CONCLUSION: Profusion of opacities on HRCT correlates with functional impairment. The presence of branching centrilobular structures may be helpful in early recognition of silicosis. PMID- 15891507 TI - Multislice computed tomography in staging lung cancer: the role of multiplanar image reconstruction. AB - OBJECTIVE: The role of multiplanar image reconstruction (MPR) in staging lung cancer was investigated using multislice helical computed tomography (CT), which allows high-quality volumetric imaging. METHODS: Forty-one consecutive patients with lung cancer (mean age = 71 years) underwent multislice CT of the thorax. The scans were acquired using contiguous 4-mm x 2.5-mm slices from the lung apex to the diaphragm in a single breath hold after injection of 100 mL intravenous contrast media. Contiguous axial, coronal, and sagittal images (5-mm slice thickness) were reconstructed in the lung and mediastinal windows. The axial images with and without multiplanar reformatted images were reviewed on a workstation on 2 separate occasions (a minimum of 6 weeks apart) by 2 experienced chest radiologists. The films were assessed for features relating to the primary lesion (size; location; and invasion of the chest wall, mediastinum, diaphragm, and/or fissures) and secondary features (mediastinal lymphadenopathy and lung metastases). The diagnostic confidence of each feature was expressed on a 4-point scale. RESULTS: A significant increase in confidence was seen on the part of both observers when diagnosing features relating to the primary lesion. The mean confidence score increased from 1.68 to 2.08 (P = 0.038) for observer A and from 1.50 to 1.80 (P = 0.020) for observer B. Confidence in assessing invasion of fissures was increased from 1.70 to 2.30 (P = 0.022) for observer A and from 1.67 to 2.27 (P = 0.006) for observer B. Improvement in interobserver agreement (kappa value from 0.61 to 0.75) was observed with multiplanar reconstruction (MPR) in the assessment of tumor location. No statistical difference was demonstrated in the diagnosis of mediastinal lymphadenopathy or lung secondaries. CONCLUSION: Multiplanar imaging of the thorax is a useful supplementary tool in the staging of lung cancer, particularly in delineating the relation of the primary lesion to fissures and the diaphragm. PMID- 15891508 TI - Mucin-producing adenocarcinoma of the lung: thin-section computed tomography findings in 48 patients and their effect on prognosis. AB - OBJECTIVE: To determine the prognostic value of thin-section computed tomography (CT) findings in patients with mucin-producing adenocarcinoma (MPA) of the lung. METHODS: The study included 48 patients with pathologically proven MPA who had thin-section CT before treatment. The CT findings were correlated with the histopathologic findings and with disease-free survival on follow-up in all patients. RESULTS: Computed tomography findings identified in patients with MPA of the lung included an air bronchogram (n = 37, 77.1%), areas of ground-glass attenuation (n = 36, 75.0%), areas of air-space consolidation (n = 36, 75.0%), interlobular septal thickening (n = 33, 68.8%), bubble-like lucencies (n = 23, 47.9%), centrilobular nodules (n = 22, 45.8%), and mucus filling of airways (n = 19, 39.6%). Twenty-two (45.8%) of the 48 patients had intrapulmonary metastases. Centrilobular nodules (odds ratio [OR] = 6.7, 95% confidence interval: 1.1-41.4; P < 0.05) and mucus filling of airways (OR = 14.4, 95% 95% confidence interval: 2.0-102.7; P < 0.01) on thin-section CT were independently associated with an increased likelihood of intrapulmonary metastases. The 5-year disease-free survival rates were 67.9% and 38.4% for patients without and with intrapulmonary metastases, respectively (P < 0.05). The presence of centrilobular nodules (relative risk = 10.5, 95% confidence interval: 1.8-59.3; P < 0.01) on thin section CT was an independent predictor of poor prognosis. CONCLUSION: Centrilobular nodules on CT are associated with a higher prevalence of intrapulmonary metastases and a poor prognosis in patients with MPA of the lung. PMID- 15891509 TI - Image-guided cutting-edge-needle biopsy of peripheral lymph nodes and superficial masses for the diagnosis of lymphoma. AB - OBJECTIVE: To evaluate the diagnostic efficacy of image-guided cutting-edge needle biopsy of peripheral lymph nodes and superficial masses for the diagnosis of lymphoma, for which many still advocate open surgical resection. METHODS: A retrospective analysis was performed of the medical records of 114 lymphoma patients who presented with peripheral lymphadenopathy and superficial masses and who underwent diagnostic image-guided biopsy. There were 69 non-Hodgkin lymphoma patients, 38 Hodgkin lymphoma patients, and 7 patients who were evaluated for histologic transformation of CLL or high grade lymphoma. RESULTS: Image-guided needle biopsy was diagnostic in 96/114 (84.2%) patients. The procedure was diagnostic in 59/69 (85.5%) of NHL patients and in 30/38 of Hodgkin disease patients (79%). Diagnoses were achieved for all 7 cases where histologic transformation was suspected. CONCLUSION: Percutaneous image-guided needle biopsy is a safe and reliable procedure with a high diagnostic yield. It can be used as a first step in patients suspected of having lymphoma presenting with enlarged peripheral lymph nodes and superficial masses. PMID- 15891510 TI - Assessment of global left ventricular function: comparison of cardiac multidetector-row computed tomography with angiocardiography. AB - OBJECTIVE: Evaluation of left ventricular function using electrocardiogram (ECG) gated multidetector row CT (MDCT) by using 3 different volumetric assessment methods in comparison to assessment of the left ventricular function by invasive ventriculography. METHODS: Thirty patients with suspected or known coronary artery disease underwent MDCT coronary angiography with retrospective ECG cardiac gating. Raw data were reconstructed at the end-diastolic and end-systolic periods of the heart cycle. To calculate the volumes of the left ventricle, 3 methods were applied: The 3-dimensional data set (3D), the geometric hemisphere cylinder (HC), and the geometric biplane ellipsoid (BE) methods. End-diastolic volumes (EDV), end-systolic volumes (ESV), the stroke volumes (SV), and ejection fractions (EF) were calculated. The left ventricular volumetric data from the 3 methods were compared with measurements from left ventriculography (LVG). RESULTS: The best results were obtained using the 3D method; EDV (r = 0.73), ESV (r = 0.88), and EF (r = 0.76) correlated well with the LVG data. The EDV volumes did not differ significantly between LVG and the 3D method (P = 0.24); however, ESV, SV, and EF differed significantly. The ESV were significantly overestimated (P < 0.01), leading to an underestimation of the SV (P < 0.01) and the EF (P < 0.01). The HC method resulted in the greatest overestimation of the volumes. The EDV and the ESV were 31.8 +/- 37.6% and 136.4 +/- 92.9% higher than the EDV and ESV volumes obtained by LVG. Bland-Altman analysis showed systematic overestimation of the ESV using the HC method. CONCLUSION: MDCT with retrospective cardiac ECG gating allows the calculation of left ventricular volumes to estimate systolic function. The 3D method had the highest correlation with LVG. However, the overestimation of the ESV is significant, which led to an underestimation of the SV and the EF. PMID- 15891511 TI - Quantitative evaluation of coronary arterial stenosis using 16-slice multidetector-row computed tomography: preliminary evaluation of phantom study. AB - OBJECTIVE: The purpose of this study is to consider the possibility of quantitative evaluation of coronary arterial stenosis by using 16-slice multidetector-row computed tomography (MDCT). METHODS: Simulated coronary arteries were prepared, which consist of 5-mm-diameter acryl tubes with contrast media (270 HU). Simulated stenoses of known density (-33 HU) were created in each coronary artery (25%, 50%, and 75%). Cardiac pulsating with 0, 50, 65, 85, and 105 beats per minute (bpm) was performed. Multiplanar reformation images for each coronary artery were created. Percent stenosis was calculated using the width middle value of boundary part of the arteries. RESULTS: The stenoses were depicted in all heart rates. Average percent stenosis +/- standard deviation was 27.4 +/- 3.6%, 45.8 +/- 2.6%, and 69.4 +/- 2.7%, respectively. For each percent stenosis, there was a significant difference (P < 0.05). CONCLUSION: Sixteen slice MDCT has a potential for noninvasive quantitative evaluation of stenosis in coronary arteries. PMID- 15891512 TI - Computed tomography colonography: automated diameter and volume measurement of colonic polyps compared with a manual technique--in vitro study. AB - OBJECTIVE: To investigate inter- and intraobserver agreement of automated measurement of polyp diameter in vitro. METHODS: Two phantoms ("QRM" and "Whiting") containing simulated polyps of known diameter and volume were scanned using 16-detector row computed tomography. Two observers estimated polyp diameter using 3 methods: software calipers ("manual"), freehand boundary identification ("semiautomatic"), and automated software segmentation ("fully automatic"). RESULTS: Intraobserver 95% limits of agreement for diameter were narrowest for the fully automatic method (QRM span: 0.39 mm, 0.48 mm; Whiting span: 0.24 mm, 0 mm). Manual estimates were approximately 10 times wider (QRM span: 3.57 mm, 3.21 mm; Whiting span: 3.2 mm, 2.02 mm). Volume estimates were narrowest for the fully automatic method (span: 24.2 mm, 24.1 mm vs. 97.9 mm, 102.9 mm for semiautomatic measurement). Interobserver agreement for diameter was narrowest for the fully automatic method (QRM span: 0.12 mm, Whiting span: 0.16 mm), with the manual method approximately 18 times wider (QRM span: 2.87 mm, Whiting span: 2.18 mm). CONCLUSION: Fully automated measurement of polyp diameter and volume is technically feasible and results in superior inter- and intraobserver agreement. PMID- 15891513 TI - Imaging after a whipple operation: improving visibility of the afferent jejunal loop in spiral computed tomography using biliary contrast medium. AB - OBJECTIVE: Spiral computed tomography (CT) examination of patients after a Whipple operation often raises questions concerning the afferent loop. This study evaluates the efficiency of biliary contrast medium (BCM) in visualizing opacification of this loop. METHODS: After undergoing a Whipple operation, 31 patients received 100 mL BCM before postoperative spiral CT (Somatom Plus 4; Siemens, Erlangen, Germany: 5- or 10-mm slices, feed of 7 or 12 mm/s; Volume Zoom; Siemens: 2.5 mm collimation, total feed of 15 mm/s). Semiquantitative evaluation included opacification of the jejunal loop and visibility of jejunostomies. Complications and tumor recurrences were recorded. RESULTS: In all patients, BCM was successfully administered without complications. Afferent loop opacification was seen in 95% of examinations with BCM versus 5% of examinations with oral contrast medium. Evaluation of jejunostomies was unequivocally facilitated by BCM. Three previously undetermined changes were identified as tumor recurrence, and recurrence was ruled out after examination with BCM in 7 cases. CONCLUSIONS: After a Whipple operation, opacification of the afferent loop can be regularly achieved by intravenous administration of BCM, which eases distinction of normal postoperative changes from local tumor recurrence. Administration of BCM is recommended as a standard diagnostic procedure in these patients. PMID- 15891514 TI - Multidetector computed tomography enteroclysis of patients with small bowel obstruction: a volume-rendered "surgical perspective". AB - Small bowel obstruction (SBO) remains a vexing clinical challenge to radiologists and surgeons. Despite advances in diagnosis, low-grade SBO can be easily missed on computed tomography (CT). Multidetector CT enteroclysis using positive oral contrast combines the volume challenge of enteroclysis with the cross-sectional multiplanar capability of multislice helical CT and allows for volume rendering of the densely opacified intestines. Such 3-dimensional models may ultimately simplify the understanding of the obstructive process as well as assist in surgical planning. PMID- 15891515 TI - Magnetic resonance imaging of maternal diseases causing acute abdominal pain during pregnancy: a pictorial review. AB - Evaluation of acute abdominal pain in a pregnant patient is a clinical challenge. In these patients, magnetic resonance imaging (MRI) can allow a systematic cross sectional evaluation of the entire abdomen and can provide clinically useful information in a short enough time for emergent diagnosis. This pictorial essay demonstrates MRI findings of various maternal diseases that can present as acute abdominal pain in pregnant patients. Familiarity with these findings is important for the radiologist to make an accurate and prompt diagnosis. PMID- 15891516 TI - Magnetic resonance experience of a twin pregnancy with a normal fetus and hydatidiform mole: a case report. AB - A case of twin pregnancy with a coexistent hydatidiform mole after in vitro fertilization is presented. Prenatal magnetic resonance (MR) imaging at 20 gestational weeks demonstrated a normal fetus and a distinct junction between the molar tissue and myometrium suggesting no evidence of myometrial invasion. Clinically, the rare disease entity involves a high risk of maternal complications and intrauterine fetal death. The application of ultrafast MR imaging for prenatal examination provides important additional information for prenatal counseling and obstetric management. PMID- 15891517 TI - Somatostatinoma of the ampulla and appendical carcinoid in a patient with von Recklinghausen disease. PMID- 15891518 TI - High-resolution flat-panel volume-CT of temporal bone--part 1: axial preoperative anatomy. AB - The purpose of this four-part series is to show the high-resolution axial and coronal anatomy of the temporal bone from a flat-panel detector-based volume CT (parts 1 and 2); these imaging planes are then used to outline the effect of different surgical procedures commonly applied to the temporal bone (parts 3 and 4). The structures that are removed or altered in 11 different surgical procedures are color-coded and inscribed in axial and coronal sections. Clinically important imaging features and complications following these surgeries will also be discussed. In these high-resolution images, many structures that are below the resolution limit of conventional CT can be seen and localized. It is hoped that one would be able to picture these structures and surgeries, in the mind's eye, even when they fall below the resolution limit using a conventional CT scanner. This article (part 1) focuses on the preoperative axial anatomy. PMID- 15891521 TI - Employees with overactive bladder: work loss burden. AB - OBJECTIVE: This study assessed the indirect work loss costs to employers as the result of employees with overactive bladder (OAB). METHODS: Study samples were drawn from an administrative database of 1.2 million beneficiaries, including medical and disability claims (1999-2002). OAB employees were compared with matched employees without OAB. RESULTS: OAB was associated with higher annual occurrences of work loss (P < 0.01). Employees with OAB had 2.2 excess work loss days as the result of medically related absenteeism and 3.4 excess days as the result of disability compared with employees without OAB (P < 0.01 for both comparisons). Employees with OAB had increased risk of disability (P < 0.01), and female employees with OAB had a higher risk of disability than male employees (P < 0.01). CONCLUSIONS: This study found that OAB was associated with work loss costs to employers resulting from increased number of employee sick days and increased risk of employee disability. PMID- 15891522 TI - Contribution of hypoglycemia to medical care expenditures and short-term disability in employees with diabetes. AB - OBJECTIVE: Diabetes is the third-most expensive physical health condition among US employees. We sought to evaluate the contribution of hypoglycemia to these costs. METHODS: We studied 2664 employees using insulin for whom medical encounters and short-term disability (STD) records were available. RESULTS: Among these employees, 442 (16.6%) had a diagnosis of hypoglycemia during an average follow-up of 2.5 years. The risk of hospitalization and emergency room visits was increased twofold in this group. Much of this excess was associated with hypoglycemia. The annualized medical cost of hypoglycemia was $3241. Patients with hypoglycemia had 77% more STD days annually. The risk of STD in the week after hypoglycemia was increased fivefold. CONCLUSION: These data suggest that hypoglycemia contributes substantially to medical care utilization and to disability-related work absence among employees using insulin. PMID- 15891523 TI - Effects of occupational trichloroethylene exposure on cytokine levels in workers. AB - OBJECTIVE: We sought to investigate trichloroethylene-induced alterations of the immune system in humans. METHODS: The levels of interleukin-2, interleukin-4, and interferon-gamma in sera obtained from workers exposed to trichloroethylene were determined and compared with those of internal and external control subjects. RESULTS: In workers with a mean urinary trichloroacetic acid concentration of 13.3 +/- 5.9 mg/g creatinine, exposed to a mean environmental trichloroethylene level of 35 +/- 14 mg/m, we observed a significant increase in sera interleukin-2 and interferon-gamma levels and a reduction in interleukin-4 concentrations compared with those of workers from the internal and external control groups. CONCLUSIONS: This study provides the first report on quantitative immune changes induced by occupational exposure to low levels of trichloroethylene and strongly suggests that exposure to this substance alters immunohomeostasis in humans with possible effects on health. PMID- 15891524 TI - Occupational exposure among bakery workers: impact on the occurrence of work related symptoms as compared with allergic characteristics. AB - OBJECTIVE: The aim of the study is to examine the relationship between exposure to occupational allergens, sensitization, and atopy and several outcome variables and the independent associations of these interrelated risk factors to work related symptoms in particular. METHODS: Data were collected by questionnaire, skin prick tests, lung function tests, and exposure measurements among 246 workers from 74 bakeries and analyzed using logistic and linear regression analyses. RESULTS: Exposure levels were associated with respiratory symptoms, sensitization to baker's allergens, and lung function. High exposure and sensitization to common and bakers' allergens were independent risk factors for work-related symptoms. However, allergy determinants performed best in predicting work-related symptoms. CONCLUSIONS: Current exposure to occupational allergens is a risk factor for work-related symptoms, but sensitization to occupational allergens and atopy are more adequate and easier obtainable predictors of work related symptoms. PMID- 15891525 TI - A cohort study of Parkinson's disease and other neurodegenerative disorders in Danish welders. AB - OBJECTIVE: We sought to evaluate rates of hospitalizations for neurodegenerative disorders in a cohort of Danish metal manufacturing employees. METHODS: A retrospective cohort study was conducted from 1977 to 2002 among 27,839 male Danish metal-manufacturing employees, with 9,817 of those employed in departments engaged in mild or stainless-steel welding and 6,163 welders. RESULTS: The standardized hospitalization ratio and 95% confidence intervals (CI) for Parkinson's disease were 0.9 (CI = 0.7-1.2) for men in steel-manufacturing companies, 1.0 (CI = 0.7-1.5) for men in welding departments, and 0.9 (CI = 0.4 1.5) for welders. Observed numbers for other neurological conditions were small and not above population expectations. Analyses for time period worked, age, and duration of welding were unremarkable. CONCLUSIONS: This relatively large cohort study with long-term follow-up provides no support for the hypothesis that rates of hospitalization for Parkinson's disease or other neurological conditions are elevated under the exposure circumstances of these Danish workers. PMID- 15891526 TI - Occupational injury among Rhode Island adolescents: an analysis of workers' compensation claims, 1998 to 2002. AB - OBJECTIVES: We sought to examine occupational injuries sustained by adolescent workers during a 5-year period. METHODS: Workers' compensation claims from Rhode Island were used to assess injuries of 15- to 19-year-old employees (n = 8321) from 1998 to 2002. Baseline employee population estimates were derived using the Current Population Survey. RESULTS: The annual estimated adolescent claim rate was 6.6 per 100 workers (95% confidence interval = 6.1-7.1), with annual average costs of $614,182. Injury rates increased with employee age, hours worked, and for day and nightshift employees relative to evening shift workers. The highest proportion of claims by industry was in eating establishments, and the average indemnification duration was 13.0 days. CONCLUSIONS: Specific emphasis should be placed on increasing safety training and preventative interventions for teenagers in hazardous occupations and for those working longer hours. PMID- 15891527 TI - Exposure and airway effects of seafood industry workers in northern Norway. AB - OBJECTIVE: In this study, we explored airway symptoms and exposure to bioaerosols and exhaust gases in seafood industry plants. METHODS: The study details the results from personal and environmental exposure measurements (17 plants), a questionnaire (n = 984), and clinical examinations (n = 225). RESULTS: The workers were exposed to allergens, endotoxins, molds, and exhaust. The 1-year prevalence of work-related airway symptoms was 42.8% for production workers and 25.9% for administrative workers. Mean levels of forced expiratory volume in 1 second and forced vital capacity were less than the predicted values in all exposed nonsmoker groups. A total of 20.5% had increased levels of total IgE (>/=100 kU/L). Specific IgE-mediated reactions seemed to be relevant only in the shrimp industry. CONCLUSIONS: Seafood industry workers showed a high prevalence of work-related airway symptoms. Further research on the relationship between exposure and effects is necessary. PMID- 15891528 TI - Stress testing in the medical evaluation for hazardous materials duty: results and consequences in three groups of candidates. AB - OBJECTIVE: The objective of this study was to determine the yield of exercise stress testing (GXT) and other methods for evaluating candidates for HAZMAT duty. METHODS: The authors conducted an analysis of prior and current records of GXTs, medical examinations, blood tests, chest radiographs, spirometry, and audiometry in 190 candidates. The authors also conducted scrutiny of GXT results, using Duke Treadmill Score (DTS), Chronotropic Index (CI), and Heart Rate Recovery (HRR). RESULTS: Seven candidates were disapproved by history and/or physical examination. Twenty-one others were deferred for GXT-induced, marked hypertension, and/or ST depression >/=2 mm. The latter appeared to be false positive indications of ischemia, low risks confirmed by DTS, CI, and HRR. Heat stress was not induced in 26 subjects so evaluated. CONCLUSIONS: GXT identified marked hypertension in 12 HAZMAT candidates and ischemic ST changes in 10, the latter appearing to be false-positives. Other testing yielded useful baselines, rarely disqualifying. PMID- 15891529 TI - Job strain, effort-reward imbalance, and heavy drinking: a study in 40,851 employees. AB - OBJECTIVE: The objective of this study was to examine the relationship of the job strain model and the effort-reward imbalance model with heavy drinking. METHODS: Questionnaire survey data were obtained from 32,352 women and 8499 men employed in the Finnish public sector (participation 67%). Logistic regression analyses for all employees and for separate subgroups were undertaken by sex, adjusted for age, education, occupational position, marital status, job contract, smoking, and negative affectivity. Different cutoff points of heavy drinking were used for men and women. RESULTS: High job strain and high effort-reward imbalance as global constructs were not associated with heavy drinking. However, some components of these models were associated with heavy drinking but the relationships were not all in the expected direction and they varied by sex, age, and occupational position. CONCLUSIONS: Stressful work conditions are not consistently associated with heavy drinking. PMID- 15891530 TI - Work-related asthma in the spray-on truck bed lining industry. AB - OBJECTIVE: The objective of this study was to identify work-related asthma (WRA) workers' compensation claims associated with methylene diphenyl diisocyanate (MDI) exposure in the spray-on truck bed lining industry and estimate the asthma incidence rate in this industry. METHODS: The authors conducted a descriptive study of workers' compensation claims meeting an established surveillance case definition for WRA. RESULTS: Eight WRA workers' compensation claims were identified in the truck bed lining industry in Washington State for a claims incidence rate of 200 per 10,000 full-time equivalent. The medical evaluation of the cases was inadequate because none of the truck bed lining cases had medical testing to objectively link their asthma to the workplace. CONCLUSIONS: The rate of work-related asthma in the truck bed lining industry is excessive and suggests a need for significant intervention, including improvements in the clinical assessment provided to MDI-exposed workers. PMID- 15891531 TI - National prevalence of asthma and chemical hypersensitivity: an examination of potential overlap. AB - OBJECTIVE: The objective of this study was to investigate the linkage between asthma and chemical hypersensitivity. METHODS: The authors conducted a population study with a random sample of 1057 geographically weighted cases to determine the prevalence of both asthma and chemical hypersensitivity in the American population and to explore their co-occurrence. RESULTS: A total of 14.1% of the respondents reported being diagnosed with asthma and 11.2% reported a hypersensitivity to chemicals. Of those with asthma, 27.2% also reported being hypersensitive to chemicals and 7.4% reported also being diagnosed with multiple chemical sensitivities (MCS). Of those diagnosed with MCS, 42% reported also being diagnosed with asthma. Additionally, 29.7% of those with asthma said air fresheners caused breathing difficulties, and 37.2% found scented products irritating. CONCLUSIONS: The results indicate that there is significant overlap between some forms of asthma and chemical hypersensitivity. PMID- 15891532 TI - Health as a complete state: the added value in work performance and healthcare costs. AB - OBJECTIVE: Health is widely believed to be more than the absence of illness, yet no previous research has documented whether organizations would benefit if occupational health moved beyond an "absence of illness" model. METHODS: Cross sectional data from the Midlife Development in the United States (MIDUS) study were used to compare productivity outcomes and health care use among individuals in (1) complete ill health, (2) incomplete health, and (3) complete health. RESULTS: Across the outcomes, individuals characterized as being completely healthy reported the greatest productivity and the lowest health care use. By contrast individuals in incomplete health had intermediate levels on outcomes and individuals in complete ill-health had the poorest outcomes. CONCLUSION: There is additional benefit of moving occupational health priorities from health as the absence of illness to health as more than the absence of illness. PMID- 15891533 TI - Worksafe--innovative web technology for occupational risk prevention. PMID- 15891534 TI - Statistics for occupational injury research: how do we measure associations and decide on statistical models? PMID- 15891537 TI - Postoperative radiographic assessment of acetabular fractures: a comparison of plain radiographs and CT scans. AB - OBJECTIVES: This investigation was designed to compare computerized tomography (CT) and plain radiography for detection of articular step and gap deformity after healing of operatively treated acetabular fractures. DESIGN: Retrospective review of CT and plain radiographic images of 15 patients treated operatively for a displaced acetabular fracture. SETTING: Level I trauma center. MAIN OUTCOME MEASURES: Ability of CT scans and plain radiographs to detect residual articular steps and gaps after healing of acetabular fractures managed by open reduction and internal fixation. RESULTS: Using standardized evaluation techniques, more patients were found to have residual articular incongruities on CT scans (8 with step and 7 with gap) than on plain radiographs (1 with step and 6 with gap). In addition, a step deformity on CT scan correlated with a gap deformity, and as the size of the step deformity increased, so too did the size of the gap deformity. CONCLUSIONS: Based on our data, computerized tomography is more likely than plain radiographs to allow detection of residual articular incongruities in healed acetabular fractures. The authors believe that CT scans improve the evaluation of articular reduction and that this information can be used to further evaluate surgical technique and provide insight as to the impact of residual articular incongruity on the development of hip arthrosis. PMID- 15891538 TI - The posterior shearing tibial plateau fracture: treatment and results via a posterior approach. AB - OBJECTIVES: This study was designed to describe the fracture patterns and early results of treatment of posterior shearing tibial plateau fractures. DESIGN: Retrospective case series. SETTING: Tertiary care hospital. PATIENTS: Thirteen patients identified from prospective trauma database with posterior shearing tibial plateau fractures. INTERVENTION: Open reduction and internal fixation through a posterior approach to the knee. MAIN OUTCOME MEASUREMENTS: Functional outcome assessed by Musculoskeletal Functional Assessment score and Visual Analogue Scale pain scores. Clinical and radiographic outcome. RESULTS: A consistent fracture pattern was identified with a primary, inferiorly displaced posteromedial shear fracture with variable amounts of lateral condylar impaction. The average duration of clinical patient follow-up was 20 (range, 13-27) months. All fractures healed after index surgery. Two complications (1 wound dehiscence and 1 flexion contracture) were all managed nonoperatively. Three independent surgeons graded patients' articular reduction, with good interobserver reliability (intraclass correlation coefficient = 0.82). The average Musculoskeletal Function Assessment dysfunction score for the 9 patients who responded was 19.5/100, and average resting Visual Analogue Scale pain score was 1.8 cm/10 cm, indicating good function. The functional outcome score was significantly related to the quality of articular reduction (P < 0.017, R = 0.456). CONCLUSIONS: Posterior shearing tibial plateau fractures form a consistent pattern. They can be successfully managed using a posterior approach with direct reduction and buttress fixation of articular fragments. Quality of articular reduction is one factor that influences short-term functional outcome. PMID- 15891539 TI - Anterior knee pain after intramedullary nailing of a tibial shaft fracture: an ultrasound study of the patellar tendons of 36 patients. AB - OBJECTIVES: Chronic anterior knee pain is a common complication following intramedullary nailing of a tibial shaft fracture. The etiology of pain is often not known. This study sonographically examined the patellar tendons of patients with a nailed tibial shaft fracture. DESIGN: Prospective study. SETTING: University hospital. PATIENTS: Fifty consecutive patients with a nailed tibial shaft fracture were initially included in the study. Thirty-six of them could be measured at an average of 2.5 +/- 0.5 years after nail insertion (1.0 +/- 0.3 years after nail extraction). INTERVENTION: Reamed intramedullary nailing with 2 interlocking bolts at both ends of the nail (Grosse-Kempf-nail, Howmedica). MAIN OUTCOME MEASUREMENTS: The ultrasound investigation of the patellar tendons of the 36 patients. RESULTS: Twelve (33%) patients were painless and 24 (67%) patients had anterior knee pain at follow-up. With the reference to the mean difference in the thickness of the distal part of the patellar tendon in the operated limb versus nonoperated limb, the result was 1.4 +/- 1.1 mm in the chronic pain group and 2.6 +/- 2.5 mm in the painless group (P = 0.135, [95% confidence interval for the group difference = -0.4-2.8]). The corresponding values for the proximal part of the patellar tendon was 1.4 +/- 1.3 mm in the chronic pain group and 2.3 +/- 2.3 mm in the painless group (P = 0.251, [95% confidence interval for the group difference = -0.7-2.4]). There were no statistically significantly differences between study groups in the blood circulation of the patellar tendon or at the entry point, calcification of the patellar tendon, granulation tissue at the entry point, or occurrence of low echo areas in the patellar tendon. CONCLUSION: After intramedullary nailing of a tibial shaft fracture, patients with or without anterior knee pain show similar changes in the ultrasound investigation of their patellar tendons. Based on those findings, it does not appear to make any difference as to the approach used (paratendinous or transtendinous) for intramedullary nailing of the tibia. PMID- 15891540 TI - Accuracy study of computer-assisted drilling: the effect of bone density, drill bit characteristics, and use of a mechanical guide. AB - OBJECTIVE: This study was designed to determine the clinical relevant accuracy of CT-based navigation for drilling. DESIGN: Experimental model. SETTING: Laboratory. METHODS: Twelve drills of varying lengths and diameters were tested with 2 different set-ups. Group 1 used free-hand navigated drilling technique with foam blocks equipped with titanium target points. Group 2 (control) used a newly developed 3-dimensional measurement device equipped with titanium target points with a fixed entry for the navigated drill to minimize bending forces. One examiner performed 690 navigated drillings using solely the monitor screen for control in both groups. The difference between the planned and the actual starting and target point (up to 150 mm distance) was measured (mm). STATISTICS: Levene test and a nonpaired t test. Significance level was set as P < 0.05. RESULTS: The core accuracy of the navigation system measured with the 3 dimensional device was 0.5 mm. The mean distance from planned to actual entry points in group 1 was 1.3 (range, 0.6-3.4 mm). The mean distance between planned and actual target point was 3.4 (range, 1.7-5.8 mm). Free-hand navigated drilling showed an increased difference with increased length of the drill bits as well as with increased drilling channel for drill bits 2.5 and 3.2 mm and not for 3.5 and 4.5 mm (P < 0.05). CONCLUSIONS: The core accuracy of the navigation system is high. Compared with the navigated free-hand technique, the results suggest that drill bit deflection interferes directly with the precision. The precision is decreased when using small diameter and longer drill bits. PMID- 15891541 TI - Alteration of body temperature, erythrocyte sedimentation rate, and C-reactive protein after reamed intramedullary nailing: a prospective study. AB - OBJECTIVES: This study was designed to confirm the clinical observation that patients undergoing reamed intramedullary nailing develop a high temperature in the early postoperative period and to further investigate this reaction by studying the alteration of biologic markers, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), during the early postoperative period. DESIGN: Body temperature, ESR, and CRP were monitored on admission, preoperatively, and postoperatively (1st, 3rd, and 10th days) in 30 consecutive patients who sustained isolated, closed femoral, or tibial diaphyseal fractures and were treated with reamed intramedullary nailing (study group). The same biologic parameters were monitored in 35 consecutive patients who sustained isolated, closed, intertrochanteric fractures of the femur at the same period of time and were treated with a Dynamic Hip Screw(R) (control group). Polytrauma patients or patients with pathologic problems or complications that could influence the results were excluded from the study. RESULTS: Body temperature, ESR, and CRP rise significantly (P < 0.0001) after reamed intramedullary nailing. CONCLUSIONS: Reamed intramedullary nailing causes significant inflammatory reaction that warrants further investigation. Knowledge that biologic markers are altered after reamed intramedullary nailing could help toward better postoperative management and undisturbed commencement of a physiotherapy program, and also toward better assessment and diagnosis of early postoperative complications, such as infection. PMID- 15891542 TI - Intertrochanteric osteotomy using a dynamic hip screw for femoral neck nonunion. AB - The dynamic hip screw, which is routinely used for intertrochanteric hip fractures, also provides a technically simple means of fixation for intertrochanteric valgus osteotomies in the treatment of femoral neck nonunions. Eight patients underwent intertrochanteric valgus osteotomy for femoral neck nonunion using a dynamic hip screw for fixation. One patient died 4 months postoperatively of causes unrelated to the procedure. The remaining 7 patients were followed for an average of 24 months postoperatively. The nonunion healed in all 7 cases. The average angle of the fracture plane decreased from 68 degrees to 41 degrees. All the hip fractures were classified as Pauwels type III preoperatively and Pauwels type II postoperatively. The average Harris Hip Score increased from 24 to 73. On radiographic review, no cases of osteonecrosis were identified postoperatively. PMID- 15891543 TI - Surgical dislocation of the hip for fractures of the femoral head. AB - Traumatic dislocations of the hip are high-energy injuries that frequently occur with fractures of the femoral head. Controversy exists regarding many aspects of the treatment of these fractures, but following reduction, large displaced head fragments require open reduction and internal fixation. Traditionally, an anterior approach to the hip joint has been used for surgical access to the femoral head, but this incision often results in limited visualization and subsequent difficulty with anatomic reduction. Recently, a surgical hip dislocation technique has been described for acetabular fractures and deformities of the proximal femur. At our institution, this technique has been used for femoral head fractures resulting in superior visualization and fracture stabilization. This article details the technique and its application. PMID- 15891544 TI - Suprascapular neuropathy complicating a Neer type I distal clavicular fracture: a case report. AB - We report the case of a 43-year-old male patient who sustained a Neer type I distal clavicular fracture after a fall onto his lateral shoulder. He was initially treated nonoperatively, and the associated suprascapular nerve injury was not diagnosed until 3 months after the accident, when the obvious hypotrophic changes in the scapular muscles were noticed. After an additional 6 weeks of observation, surgery was performed due to the absence of any clinical and electrodiagnostic evidence of recovery. At the time of exploration, through a posterior approach, the suprascapular nerve was found to be entrapped in scar tissue located proximally to the suprascapular notch. A surgical release of the transverse scapular ligament and external neurolysis of the embedded scar tissue was done. One month after the procedure, the patient could effectively elevate his arm to 180 degrees and externally rotate his shoulder to 75 degrees . The last muscle examination (1 year postsurgery) demonstrated grade 5 abductors and grade 4 external rotators, but the muscle bulk had not returned to normal size at 1.5 years after operation. Our experience with this case emphasizes the importance of serial examinations of the shoulder following trauma to that joint. PMID- 15891545 TI - Diagnosis of thoracic neurilemoma incidental to a traumatic clavicle fracture. AB - This report highlights the importance of thorough examination of the complete radiograph in a fracture setting. PMID- 15891546 TI - Avulsion fractures of the lesser tuberosity of the humerus in adolescents: review of the literature and case report. AB - Isolated fracture of the lesser tuberosity is an unusual phenomenon in children and adolescents. These injuries are difficult to diagnose acutely and often present as chronic shoulder pain. In this study, we report on 1 case of a displaced lesser tuberosity apophysis avulsion fracture in an adolescent treated with open reduction and internal fixation, as well as a review of the literature. A 14-year-old adolescent male presented to the senior surgeon complaining of left shoulder pain and weakness 10 days after a wrestling injury. He was diagnosed with a displaced, isolated fracture of the lesser tuberosity apophysis for which he underwent open reduction and internal fixation. A combination of sutures passed through drill holes in the proximal humerus and bioabsorbable suture tacks were used to anatomically fix the lesser tuberosity fragment and subscapularis tendon. Postoperatively, he underwent a progressive physical therapy regimen. At 4 months follow-up, he had full range of motion, complete return of strength, and returned to competitive athletics. We report here on the successful surgical treatment of a fracture of the lesser tuberosity apophysis in an adolescent. PMID- 15891547 TI - Necrotizing infection due to Bacillus cereus mimicking gas gangrene following penetrating trauma. AB - An 8-year-old boy presented with fulminant necrotizing infection resembling gas gangrene following penetrating trauma from a tree branch. Bacillus cereus was isolated from tissue specimens, showing that unexpected pathogens can be isolated. It is essential to submit specimens for culture, as this organism is typically resistant to beta-lactam antibiotics and metronidazole, the empiric choice for gas gangrene. PMID- 15891548 TI - Calcaneal fractures: to fix or not to fix: Opinion: open reduction internal fixation. PMID- 15891549 TI - Calcaneal fractures: to fix or not to fix: Opinion: nonoperative approach. PMID- 15891550 TI - Displaced intra-articular calcaneal fractures. AB - BACKGROUND: Calcaneal fractures comprise 1 to 2 percent of all fractures. Approximately 75% of calcaneal fractures are intra-articular. The management of intra-articular calcaneal fractures remains controversial. Nonoperative treatment options include elevation, ice, early mobilization, and cyclic compression of the plantar arch. Operative treatment options include closed reduction and percutaneous pin fixation, open reduction and internal fixation, and arthrodesis. The effect of operative versus nonoperative treatment has been the focus of several comparative studies. OBJECTIVE: This study was designed to determine the effect of operative treatment compared with nonoperative treatment on the rate of union, complications, and functional outcome after intra-articular calcaneal fracture in adults. PMID- 15891551 TI - Gender differences in patients with hip fracture: a greater risk of morbidity and mortality in men. PMID- 15891553 TI - A modified judet approach to the scapula. PMID- 15891555 TI - L-asparaginase: old dog, more tricks and leukemia: not simply a liquid tumor. PMID- 15891556 TI - Pyrite or true gold? PMID- 15891557 TI - New insights on asparaginase. PMID- 15891558 TI - Cross-sectional study of bone mineral density in adult survivors of solid pediatric cancers. AB - To investigate the hypothesis that survivors of pediatric solid cancer have low bone mineral density, a cross-sectional study was done of subjects who had received treatment for pediatric solid tumors before 16 years of age and were less than 40 years old at follow-up. Excluded were subjects treated for acute lymphoblastic leukemia or those who had received cranial irradiation, total body radiation, or nonautologous bone marrow transplant. The study group consisted of 38 subjects, with the most common diagnoses being lymphoma (n = 17), sarcoma (n = 8), Wilms tumor (n = 5), and neuroblastoma (n = 4). Median age was 22 years (range 12-32). Time from diagnosis of underlying cancer averaged 12.6 years (range 5.5-20.3). Using criteria of osteopenia (Z-score < or = -1.0 and > -2.0) and osteoporosis (Z-score < or = -2.0) for any one or more areas including total body, lumbar spine, total hip, or femoral neck density, 13 of the 38 subjects (34%) had osteopenia or osteoporosis. A further six subjects (16%) had isolated upper extremity osteopenia or osteoporosis. Multivariate analysis showed a direct relationship between the number of chemotherapy drugs administered and the presence of osteopenia or osteoporosis in the lower extremities (P = 0.03). Young survivors of childhood solid tumors are at increased risk of developing premature osteopenia or osteoporosis, and screening evaluations and follow-up are warranted. PMID- 15891559 TI - Temozolomide and radiation for aggressive pediatric central nervous system malignancies. AB - This study describes the outcomes of children treated with combinations of temozolomide and radiation therapy for various aggressive central nervous system malignancies. Their age at diagnosis ranged from 1 to 15 years. Patients with focal disease were treated with concomitant temozolomide (daily 75 mg/m) and three-dimensional conformal radiotherapy in a dose that ranged from 50 to 54 Gy, followed by temozolomide (200 mg/m/d x 5 days/month in three patients, 150 mg/m x 5 days/ month in one patient). Patients with disseminated disease were treated with craniospinal radiation (39.6 Gy) before conformal boost. One patient received temozolomide (200 mg/m x 5 days/month) before craniospinal radiation, and one patient received temozolomide (daily 95 mg/m) concomitant with craniospinal radiation and a radiosurgical boost, followed by temozolomide (200 mg/m x 5 days/month). Three patients achieved a partial response during treatment, with two of these patients dying of progressive disease after treatment. One patient has no evidence of disease. Three patients achieved stable disease, with one of these patients dying of progressive disease after treatment. Toxicities observed included low-grade neutropenia, thrombocytopenia, and lymphopenia. The combination of temozolomide and radiotherapy appears to be well tolerated in a variety of treatment schemas for aggressive pediatric central nervous system malignancies. This information is of particular use in designing future studies, given the recent positive results in a randomized study examining the use of temozolomide concomitant with radiation in the treatment of adult glioblastoma. PMID- 15891560 TI - Aseptic osteonecrosis in children and adolescents treated for hemato-oncologic diseases: a 13-year longitudinal observational study. AB - Aseptic osteonecrosis (AON) is a serious long-term complication of childhood cancer therapy. A retrospective study was undertaken to describe treatment and long-term follow-up of patients with AON. Between 1990 and 2003, 630 consecutive children with various malignancies were treated at the University Children's Hospital in Graz, Austria. In nine of these patients presenting with skeletal symptoms, MRI revealed AON. All nine had hematologic malignancies. The median age at diagnosis of malignancy was 15.8 years (range 13.7-18.6 years), and the median interval between diagnosis of malignancy and onset of osteonecrosis-related symptoms was 16 months (range 6-53 months). All patients had received previous corticosteroid therapy. Treatment of AON included restriction of weight-bearing, physiotherapy, and analgesics. Three patients were treated with hyperbaric oxygen therapy combined with the prostacyclin analog iloprost, and one patient also received pamidronate, a second-generation bisphosphonate. This conservative treatment resulted in alleviation of symptoms in all patients. One patient had to undergo bilateral hip replacement and two had to undergo arthrotomy with sequestrotomy due to subsequent deterioration of symptoms. Close monitoring for skeletal symptoms is mandatory during follow-up of patients with hematologic malignancies. Previous corticosteroid treatment and age older than 10 years seem to be major risk factors. Early detection of AON leading to prompt intervention may prevent more severe morbidity. PMID- 15891561 TI - Recurrent central nervous system medulloepithelioma: response and outcome following marrow-ablative chemotherapy with stem cell rescue. AB - Medulloepithelioma is a rare primitive neuroectodermal tumor of the central nervous system usually developing in childhood, displaying highly malignant behavior, with early progression or recurrence. Once a recurrence develops, death reportedly follows with invariable rapidity. The purpose of this study was to evaluate the efficacy of high-dose, marrow-ablative chemotherapy with autologous hemopoietic stem cell rescue in the treatment of recurrent central nervous system medulloepithelioma. Three young children with recurrent central nervous system medulloepithelioma received high-dose marrow-ablative chemotherapy with thiotepa and etoposide either alone (one patient) or with the addition of carboplatin (two patients). This was followed by irradiation only in one patient. One child with residual radiographic tumor at the time of treatment could be evaluated for response and showed complete resolution of leptomeningeal disease after receiving marrow-ablative chemotherapy. Two children developed tumor recurrence at 2.0 and 5.5 months after receiving marrow-ablative chemotherapy. The third child continues free of tumor beyond 12 years from treatment. The authors' experience with marrow-ablative chemotherapy and autologous hemopoietic stem cell rescue suggests that this treatment strategy might be beneficially incorporated into the initial treatment approach for young children with medulloepithelioma. PMID- 15891562 TI - Cavoatrial tumor extension in children with wilms tumor: a retrospective review of 17 children in a single center. AB - The aim of this study was to evaluate the clinical characteristics and treatment results of 17 children with cavoatrial tumor extension of Wilms tumor. Of the 360 Wilms tumors diagnosed between 1980 and 2000, 17 patients with intracaval thrombus were identified from the medical records at the pediatric oncology department of Hacettepe University. The following data were collected and reviewed: age, sex, presenting symptoms, tumor site, presence of anaplasia, stage, associated congenital anomalies, localization of tumor thrombus, radiologic findings, type and duration of preoperative chemotherapy, response to preoperative chemotherapy, recurrences, and survival. The frequency of cavoatrial extension in this group was 4.7% (15 in the inferior vena cava and 2 in the right atrium). Fourteen patients received preoperative chemotherapy consisting of two drug regimen (vincristine and actinomycin D) ranging from 1 to 12 weeks (median 4 weeks). Since intravascular invasion is often asymptomatic, a careful radiologic examination to detect tumor thrombus before surgery is essential. There is no need for aggressive surgery in the presence of tumor thrombus. It may be resolved by preoperative chemotherapy. Surgical removal of the thrombus should be considered in the presence of life-threatening tumor thrombosis at diagnosis and in patients who had residual thrombus after chemotherapy. PMID- 15891563 TI - Myelopathy due to intrathecal chemotherapy: report of six cases. AB - Intrathecal chemotherapy and systemic chemotherapy are used for both prophylaxis and treatment of central nervous system disease in hematologic malignancies. However, intrathecal treatment has some adverse effects, such as arachnoiditis, progressive myelopathy, and leukoencephalopathy. The authors describe six children in whom myelopathy and adhesive arachnoiditis developed after administration of intrathecal chemotherapy including methotrexate, cytosine arabinoside, and prednisolone. Urinary retention and incontinence, the main presenting complaints in all patients, developed within 12 hours after intrathecal therapy and spontaneously resolved within 7 days. Two patients were unable to walk. In these two, weakness in the lower extremities gradually recovered by 1 month but urinary incontinence did not improve. None of the children had sensory loss. On follow-up periodic recurrent urinary tract infection was noted in four patients. MRI findings corresponded to arachnoiditis. No response was recorded on tibial nerve somatosensory evoked potentials in all patients. Intrathecal chemotherapy, especially methotrexate, can cause spinal cord dysfunction in children with acute lymphoblastic leukemia and non-Hodgkin's lymphoma. Arachnoiditis should be kept in mind as a causative factor in recurrent urinary tract infection in patients receiving intrathecal chemotherapy. PMID- 15891564 TI - Antibody titers and immune response to diphtheria-tetanus-pertussis and measles mumps-rubella vaccination in children treated for acute lymphoblastic leukemia. AB - The objective of this study was to investigate the diphtheria-tetanus-pertussis and/or measles-mumps antibody titers before and after vaccination at various time points of acute lymphoblastic leukemia (ALL) therapy and to suggest an appropriate vaccination approach for ALL patients. The authors studied 37 ALL patients and 14 healthy control subjects, divided into three groups. In group 1 (newly diagnosed patients), baseline anti-diphtheria, anti-tetanus, and anti pertussis titers were determined. Patients in group 2 (on maintenance chemotherapy) and group 3 (patients not receiving therapy for 3-6 months) were vaccinated with diphtheria-tetanus with or without acellular pertussis; group 3 and control subjects were also given measles-mumps-rubella vaccine. Preimmunization and 1-month postimmunization titers were drawn. Preimmunization anti-diphtheria and anti-tetanus antibody titers between the groups and the controls were statistically similar. The seropositivity rate for anti-measles antibody in group 3 was significantly lower than controls. After vaccination, all of the patients developed protective anti-diphtheria and anti-tetanus antibody titers. The seroconversion rates of group 3 and controls for anti-measles and anti-mumps antibodies were statistically similar. The results showed that patients on maintenance therapy and after cessation of therapy made good antibody responses to diphtheria and tetanus toxoids, but response to measles and mumps vaccines was not as sufficient as toxoid vaccines. Children with ALL can receive the appropriate vaccines during and after maintenance treatment. PMID- 15891565 TI - Idiopathic myelofibrosis with generalized periostitis in a 4-year-old girl. AB - Idiopathic myelofibrosis, a chronic myeloproliferative disorder of unknown origin, is characterized by splenomegaly, extramedullary hematopoiesis, leukoerythroblastosis, teardrop erythrocytes, and myelofibrosis. It is a rare disorder in childhood. The authors describe a 4-year-old girl with features consistent with idiopathic myelofibrosis, who also had generalized solid laminated periosteal reaction involving all long bones. The presence of thrombocytopenia at the onset and lack of leukocytosis were in contrast to the reported features seen in children. Recent case reports describe a relatively indolent course in children. Spontaneous remissions have also been described in pediatric cases. The fulminant course of this patient without any features of malignant transformation was noteworthy in this regard. PMID- 15891566 TI - Fungemia due to Trichosporon asahii in a neutropenic child refractory to amphotericin B: clearance with voriconazole. AB - Disseminated Trichosporon infection in neutropenic patients carries a poor prognosis. Clinical evidence on the use of voriconazole for this infection is limited. The authors report a case of Trichosporon asahii fungemia refractory to liposomal amphotericin B treatment in a boy with acute lymphoblastic leukemia, which resolved after the addition of voriconazole. Both voriconazole and amphotericin B exhibited low minimal inhibitory concentrations and minimal fungicidal concentrations, and their combination was indifferent in vitro. The use of voriconazole for the treatment of trichosporonosis in patients with hematologic malignancies deserves further study. PMID- 15891567 TI - Vincristine-induced urinary bladder paralysis. AB - The authors describe the development of bladder paralysis in a child with acute lymphoblastic leukemia undergoing maintenance chemotherapy. Immediately before the adverse clinical event, the child had received vincristine intravenously and triple therapy with hydrocortisone, cytosine arabinoside, and methotrexate intrathecally and had begun a 5-day pulse of prednisolone. The authors conclude that the ensuing reversible bladder paralysis was related to the vincristine. The clinical event resolved, and vincristine was deleted from the child's subsequent therapy until full recovery was achieved. The authors advise recognition of this problem and discontinuation of the vincristine if transient bladder paralysis develops until symptoms completely disappear. PMID- 15891568 TI - Life-threatening hypercalcemia complicated by pancreatitis in a child with acute lymphoblastic leukemia. AB - The authors describe a 9-year-old girl with precursor-B acute lymphoblastic leukemia (ALL) who presented with dehydration and severe hypercalcemia. She had received oral vitamin D and calcium supplementation for 4 days, the last dose 48 hours prior to admission, and required pediatric intensive care unit (PICU) hospitalization for management of the hypercalcemia and safe initiation of induction chemotherapy. Her clinical course was complicated by pancreatitis, disseminated intravascular coagulation, pleural effusion, and focal seizures. Although the exact mechanism of hypercalcemia was not elucidated, it was likely related to the underlying ALL, without dismissing the prior vitamin D and calcium supplementation as a possible contributing factor. The hypercalcemia resolved with specific antileukemic therapy along with supportive care and administration of calcitonin. Hypercalcemia is an uncommon metabolic abnormality in children with ALL, but it can be life-threatening. Children with ALL should be referred to tertiary-care institutions with PICU and subspecialty support because serious metabolic and other complications can occur before or after the administration of chemotherapy. PMID- 15891569 TI - Relapse of Wilms' tumor 13 years after original diagnosis. PMID- 15891570 TI - Innervation of stem cell-derived neurons into auditory epithelia of mice. AB - This study examined the potential of embryonic stem cell-derived neurons as transplants for cell therapy for the primary loss of spiral ganglion neurons. To assess the ability of embryonic stem cell-derived neurons for innervation into auditory epithelia, they were cocultured with auditory epithelium explants of mice for 7 days. Histological analysis demonstrated massive elongation of neurites from embryonic stem cell-derived neurons toward auditory hair cells. Embryonic stem cell-derived neurites were adjacent to or surrounding hair cells, and exhibited expression of synaptophysin, a marker for synaptic vesicles. These findings demonstrate the ability of embryonic stem cell-derived neurons for reinnervation into auditory epithelia, indicating a high potential of embryonic stem cell-derived neurons as transplants for replacing spiral ganglion neurons. PMID- 15891571 TI - Leftward asymmetry in relative fiber density of the arcuate fasciculus. AB - Left hemispheric language dominance is well established, but the structural substrate for this functional asymmetry is uncertain. We report a strong asymmetry in the relative fiber density of the arcuate fasciculus, a white matter pathway associated with language that connects the frontal, temporal, and parietal lobes. Measured with diffusion tensor tractography, nearly all study participants demonstrated greater relative fiber density in the left arcuate fasciculus than in the right arcuate fasciculus. In comparison, we found no asymmetry in the corticospinal tract, an important white matter pathway with no known role in language. Combined with data on volumetric and activation asymmetry, greater connectivity may provide the elements of a neural system model for language lateralization. PMID- 15891572 TI - Sex differences in the human corpus callosum: diffusion tensor imaging study. AB - In order to assess underlying structural differences between the male and female corpus callosum, the fractional anisotropy and volume of the corpus callosum, and also its T1 signal intensity, were measured. The corpus callosum of the 15 normal women and 15 normal men was drawn on the mid-sagittal T1-weighted image, for determining its volume and signal intensity, and this region of interest was projected onto the coregistered fractional anisotropy image, in order to obtain the value for the corpus callosum. We found increased T1 signal intensity and decreased fractional anisotropy in the female corpus callosum, as compared with that of the male. Despite the long-standing debates, the corpus callosum remains a region of sex differences. PMID- 15891573 TI - Transient cochlear ischemia and its effects on the stria vascularis. AB - The effects of transient cochlear ischemia on the stria vascularis were studied. Fifteen minutes of ischemia decreased the endocochlear potential by up to 17.5 mV on day 1; it returned to normal on day 7. Immunostaining for Na+,K+-ATPase, a marker for the Na+/K+-pump, and for connexin 26, a marker for gap junctions, was inhibited on days 1 and 4, and returned to normal on day 7. Electron microscopy showed expansion of the intercellular space with abundant vacuolar formation in the stria vascularis. These morphological changes disappeared completely by day 7. The results indicate that transient ischemia causes a reversible functional disorder of the stria vascularis with fine structural changes, which may be owing to dysfunction of Na+/K+-pump or gap junctions. PMID- 15891574 TI - Audiovisual phonological mismatch produces early negativity in auditory cortex. AB - During silent reading, visual information provided by letters is converted to auditory information in the mind. The purpose of this study was to identify the primary locus for auditory verbal imagery in the brain. Neuromagnetic recording was obtained from 10 right-handed study participants, who were instructed to identify infrequently occurring phonological mismatches between a random-ordered sequence of syllable sounds and a visually presented syllabogram sequence. The activity difference in early latency, calculated by subtracting the averaged responses to matched syllables from the averaged responses to mismatched syllables, showed a spatiotemporal profile strikingly similar to that of mismatch negativity. Auditory imagery of forthcoming verbal sounds may establish a memory trace as a template for imagery-based mismatch negativity generation in the auditory cortex. PMID- 15891575 TI - Transmission of neural activity in a feedforward network. AB - In this work, the enhancement of coherence resonance of firings in a 10-layer feedforward neuronal network with sparse couplings is found when there is noise input to each layer. Periodic signals with frequency 30-80 Hz are found to be well transmitted though the network, and such a frequency sensitivity can be modulated by the noise intensity and is different in different layers. When a random pulse-like signal is input to the neurons of the first layer, the signal can be well read out from the population rates in an optimal range of noise intensity. This ability decreases as the layer index increases. PMID- 15891576 TI - Sodium salicylate reduces gamma aminobutyric acid-induced current in rat spinal dorsal horn neurons. AB - Sodium salicylate is one of the nonsteroidal antiinflammatory drugs and is clinically used for antiinflammation and chronic pain relief. In the present study, we investigated the actions of sodium salicylate on gamma-aminobutyric acid type A receptor (GABA(A)) current in cultured rat spinal dorsal horn neurons. Sodium salicylate was found to reduce GABA(A) current in a reversible and concentration-dependent manner, but did not change its ion selectivity. Sodium salicylate was effective only when GABA and sodium salicylate were applied together. Application of sodium salicylate immediately before, but not during, the application of GABA did not result in a significant reduction of GABA(A) current. Our results demonstrate that sodium salicylate reversibly attenuates the GABA(A) response of dorsal horn neurons, suggesting that GABA(A) receptors in the region are pharmacological targets of sodium salicylate. PMID- 15891577 TI - Independence of visual awareness from attention at early processing stages. AB - According to a widely accepted idea, only the results of attentional selection reach visual awareness. A competing model postulates that awareness is independent of attentional selection: contents of subjective visual experience may also exist without the contribution of attention. We tested these competing models by tracking the independent contributions of selective attention and awareness to electrophysiological brain responses. Our results showed that the earliest effects of visual awareness emerged earlier than the effects of attention and regardless of the presence or absence of attention. The early effects of attention were elicited regardless of the presence or absence of awareness. The results suggest that visual awareness and selective attention are initially independent of each other. PMID- 15891578 TI - Activation of c-Jun N-terminal kinase is required for neurite outgrowth of dopaminergic neuronal cells. AB - Recent studies indicate that activation of stress-activated protein kinases may be implicated in a broad range of biological activities including differentiation. To directly examine whether stress-activated protein kinases are involved in neuronal differentiation, we utilized retinoic acid-induced and spontaneous models of neurite outgrowth in dopaminergic neurons. Here, we show that retinoic acid-induced neurite outgrowth in MN9D dopaminergic neuronal cells was accompanied by activation of c-Jun N-terminal kinase but not p38. Consequently, cotreatment with a specific inhibitor of c-Jun N-terminal kinase or overexpression of c-Jun N-terminal kinase-binding domain of c-Jun N-terminal kinase-interacting protein-1 blocked retinoic acid-induced neurite outgrowth. In primary cultures of dopaminergic neurons, the extent of neurite outgrowth increased spontaneously in a time-dependent manner. When these cultures were treated with a specific inhibitor of c-Jun N-terminal kinase, the total extent of neurites, the primary neurite length and the number of neurites per cell were suppressed significantly. Thus, our data indicate that the c-Jun N-terminal kinase signal seems to play an important role during morphological differentiation in cultured dopaminergic neurons. PMID- 15891579 TI - PPAR-alpha activators suppress STAT1 inflammatory signaling in lipopolysaccharide activated rat glia. AB - Signal transducers and activators of transcription (STATs) have recently been reported to mediate glial activation, and thus potentially play important roles in many neuroinflammatory diseases. We examined the effect of peroxisome proliferator-activated receptor (PPAR) activators on inflammatory responses in cultured rat brain glial cells. Four PPAR-alpha activators were tested, three fibrates (WY14643, clofibrate and fenofibrate) and an arachidonic acid derivative (5,8,11,14-eicosatetraynoic acid). We found that all four PPAR-alpha activators suppressed lipopolysaccharide-stimulated STAT1 phosphorylation and nuclear factor binding to gamma-interferon-activated sequence/interferon-alpha-stimulated response element sites known to contain STAT binding sites. PPAR-alpha activators also suppressed lipopolysaccharide-stimulated tumor necrosis factor-alpha and monocyte chemoattractant protein-1 transcription and release. These results suggest that PPAR-alpha activators may be useful in the treatment of inflammatory brain diseases. PMID- 15891580 TI - Lack of response to epidermal growth factor in adult neural progenitor cells. AB - Neural progenitor/stem cells reside in the subventricular zone and dentate gyrus of the adult rat brain, which proliferate in response to several growth factors. Here we show that the neurospheres generated from the adult dentate gyrus and subventricular zone proliferate in vitro in response to fibroblast growth factor 2, but not epidermal growth factor. Likewise, extracellular signal-regulated kinase phosphorylation was stimulated by fibroblast growth factor-2, but not epidermal growth factor. Immunoblotting demonstrated negligible epidermal growth factor receptor content in these neurospheres, indicating that neural progenitor cells isolated from the adult rat brain are not responsive to epidermal growth factor in vitro, because of the lack of epidermal growth factor receptors and/or their signaling. However, the lack of response by epidermal growth factor in our study could be because of the culture conditions, and may not reflect the physiological condition. PMID- 15891581 TI - Age-dependent effects of the 5-hydroxytryptamine-2a-receptor polymorphism (His452Tyr) on human memory. AB - A polymorphism (His452Tyr) of the 5-hydroxytryptamine (5-HT)2a receptor is associated with episodic memory in healthy young humans. Because 5-HT2a-receptor density decreases with increasing age, we tested whether the 5-HT2a receptor genotype effect on memory is influenced by age. We investigated the association of the His452Tyr genotype with memory performance in 622 healthy study participants aged from 18 to 90 years. In young to middle-aged participants, age significantly influenced genotype effects on episodic memory: the His452Tyr genotype exerted a significant influence on memory only in young participants. In the group of elderly cognitively healthy participants, the His452Tyr genotype did not affect memory performance. We conclude that age strongly modulates the effect of the 5-HT2a receptor polymorphism at residue 452 on episodic memory. PMID- 15891582 TI - Visuomotor links in awareness: evidence from extinction. AB - In patients with extinction, ipsilesional stimuli may abolish awareness of contralesional stimuli. Explanations of extinction often assume a serial model of processing in which sensory competition and identification precedes the selection of responses. We tested the adequacy of this assumption by examining the effects of response variables on visual awareness in six patients using signal detection analysis. Ipsilesional stimuli modulated patients' response criteria in deciding whether a contralesional stimulus was a target, and response modality (verbal or motor) modulated patients' abilities to discriminate between contralesional targets and distractors. This pattern of input variables modulating response criteria and output variables modulating discriminability indicates the extent to which attentional and intentional systems are tightly intertwined, with bi directional effects in producing visual awareness. PMID- 15891583 TI - Lateralized predatory responses in the ornate dragon lizard (Ctenophorus ornatus). AB - Lateralized responses for visually elicited feeding behaviour have been reported in toads and birds but not in the phylogenetically intermediate class of vertebrates, the reptiles. Here we examined small social groups of ornate dragon lizards Ctenophorus ornatus (family Agamidae) and provide the first report in reptiles of right eye lateralization (left brain hemisphere) for predatory responses to prey. However, right eye lateralization was not evident initially but became stronger with time supporting a shift to right eye lateralization as the prey became increasingly familiar. The study is in agreement with recent findings in toads, adding credence to the hypothesis that lateralization originated in an early ancestor and highlighting the supposition that the strength and direction of lateralization is dependent on experience. PMID- 15891584 TI - Rapid eye movement-related brain activation in human sleep: a functional magnetic resonance imaging study. AB - In animal models, ponto-geniculo-occipital waves appear as an early sign of rapid eye movement sleep and may be functionally significant for brain plasticity processes. In this pilot study, we use a combined polysomnographic and functional magnetic resonance imaging approach, and show distinct magnetic resonance imaging signal increases in the posterior thalamus and occipital cortex in close temporal relationship to rapid eye movements during human rapid eye movement sleep. These findings are consistent with cell recordings in animal experiments and demonstrate that functional magnetic resonance imaging can be utilized to detect ponto-geniculo-occipital-like activity in humans. Studying intact neuronal networks underlying sleep regulation is no longer confined to animal models, but has been shown to be feasible in humans by a combined functional magnetic resonance imaging and electroencephalograph approach. PMID- 15891585 TI - Body mass predicts orbitofrontal activity during visual presentations of high calorie foods. AB - Little is known about the relationship between weight status and reward-related brain activity in normal weight humans. We correlated orbitofrontal and anterior cingulate cortex activity as measured by functional magnetic resonance imaging with body mass index in 13 healthy, normal-weight adult women as they viewed images of high-calorie and low-calorie foods, and dining-related utensils. Body mass index correlated negatively with both cingulate and orbitofrontal activity during high-calorie viewing, negatively with orbitofrontal activity during low calorie viewing, and positively with orbitofrontal activity during presentations of nonedible utensils. With greater body mass, activity was reduced in brain regions important for evaluating and modifying learned stimulus-reward associations, suggesting a relationship between weight status and responsiveness of the orbitofrontal cortex to rewarding food images. PMID- 15891586 TI - Preattentive interference between touch and audition: a case study on multisensory alloesthesia. AB - Alloesthesia is a rare clinical condition that corresponds to a spatial disorder of stimulus localization, in which patients experience a given stimulus on the side opposite to the side of stimulation. Whereas it has been mostly described for unisensory stimulations, evidence of multisensory alloesthesia is only anecdotal. Here, we investigated a case of multisensory auditory-tactile alloesthesia. Our data suggest that auditory-tactile integration and multisensory alloesthesia not only depend on attentional mechanisms, but also on somatotopic preattentive mechanisms. PMID- 15891587 TI - Prechange event-related potentials predict change blindness in various attention conditions. AB - We investigated the mechanisms of change blindness by measuring electrophysiological responses in humans to the prechange display. The change blindness paradigm included different attentional cueing (valid, invalid, no cue) conditions. The results showed that event-related potentials to the prechange display predicted change detection performance. After a valid cue, the prechange responses in detected change trials showed evidence of attentional focusing. However, in no cue and invalid cue conditions successful change detection was associated with electrophysiological responses indicating more globally distributed attention, whereas the electrophysiological pattern associated with strong attentional focusing predicted change blindness. The results suggest that specific patterns of encoding the prechange display predict change detection performance not only between different attention conditions but also within them. PMID- 15891588 TI - When case meets agreement: event-related potential effects for morphology-based conflict resolution in human language comprehension. AB - We present event-related potential evidence from language comprehension that processing conflicts arising from the same linguistic domain and appearing within the same time range do not interact when they draw upon distinct underlying neural populations. Thus, a combined violation of two morphosyntactic information types, number-agreement and case, engendered a LAN/N400-P600 pattern, while the corresponding single violations are associated with LAN-P600 and N400-P600 responses, respectively. The absence of an interaction between the two negativities indicates that neuronal resource sharing does not result from a similarity of function, but rather requires an overlap of the underlying neuronal populations. PMID- 15891589 TI - Tritium dosimetry. PMID- 15891590 TI - Large variations occur in bone density measurements of children when using different software. AB - BACKGROUND: Paediatric dual X-ray absorptiometry (DXA) studies present a number of technical problems. One of these is that the edge detection algorithms designed for the adult skeleton may fail for paediatric studies. Hologic provide alternative algorithms for low bone density studies. AIM: To assess low-density software for the analysis of paediatric DXA studies and to compare with the adult protocol. METHODS: Our centre has scanned 450 normal children as part of a normal range study. A subgroup of 103 children was selected using a random number generator. The group was distributed evenly between males and females and across the age range 5-17 years. Each individual underwent both a lumbar spine and a whole-body scan on a Hologic QDR-4500W DXA scanner. Both scans were analysed using the standard adult protocol and then re-analysed using the Hologic experimental paediatric protocol for whole body and the Hologic low-density protocol for lumbar spine. RESULTS: Both lumbar spine protocols showed an increase in bone mineral density with age; however, the low-density protocol always produced a lower bone mineral density result than the adult protocol. Bland-Altman analysis showed limits of agreement of 0.031-0.093 g x cm(-2) (male, 0.032-0.089 g x cm(-2); female, 0.031-0.096 g x cm(-2)). This represents a mean difference of 9%. Five results showed differences greater than the upper limit of agreement. All these cases were children under 11 years of age who had large areas of spine not identified as bone by the adult protocol. These children were all below the 30th percentile for the body mass index. The whole-body protocols showed similar increases in bone mineral density with age; however, the experimental paediatric protocol always produced a lower bone mineral density result than the adult protocol. Paired results showed limits of agreement of 0.0668-0.130 g x cm(-2) (male, 0.063-0.124 g x cm(-2); female, 0.073-0.134 g x cm(-2)). This represents a mean difference of 11%. Five results showed differences greater than the upper limit of agreement. CONCLUSIONS: For anteroposterior (AP) lumbar spine scans, the use of the paediatric algorithm in children under 11 years of age would prevent the largest failures in analysis. For whole-body scanning, the adult algorithm showed no major failures in children of 11 years or older. It is hoped that forthcoming improvements in whole-body density analysis will improve the results for those under 11 years of age. Normal range data should be generated for any new algorithm to allow proper interpretation of clinical studies. PMID- 15891591 TI - Role of 99mTc Sulesomab in the diagnosis of prosthetic joint infections. AB - BACKGROUND: Investigation of suspected infection in prosthetic joint replacements is a difficult orthopaedic problem with important therapeutic implications. METHODS: We retrospectively analysed the results of 38 patients referred with a clinical suspicion of bone or joint infections. Referrals were made for suspected infection of prosthetic hip joints (17), total knee replacements (13), total elbow replacements (4) and total shoulder replacements (4). Sulesomab imaging was done with 650 MBq of 99mTc Sulesomab. The scintigraphic diagnosis was compared with the final clinical diagnosis and information collected from routine blood tests (erythrocyte sedimentation rate, C-reactive protein and full blood count), plain radiographs, appropriate microbiology, culture and/or histology where available. The final diagnosis was determined by conclusive microbiology, culture and/or histology, intra-operative findings, aspiration, complementary investigations like computed tomography/magnetic resonance imaging and long-term clinical follow-up. RESULTS: The overall sensitivity was 90.90% and specificity 81.48%. The individual sensitivity and specificity of each category were compared. Knee prostheses showed better sensitivity and specificity compared to that for hip prostheses. CONCLUSIONS: 99mTc Sulesomab seems to be useful in excluding infection rather than confirming it, with a high negative predictive value (95.65%). PMID- 15891592 TI - Age related preservation and loss in optimized brain SPECT. AB - BACKGROUND: Recent single photon emission computed tomography (SPECT) studies have reported age related increases in regional brain perfusion (called preservation here) as well as losses. AIM: To apply optimized SPECT processing to better define and understand both age related preservation and loss in brain SPECT. METHODS: Brain SPECT was performed on 85 healthy subjects using Tc hexamethylpropylene amine oxime (HMPAO), processed using findings from recent optimization work, and subjected to voxel based statistical analysis. RESULTS: SPECT preservation was seen in white matter. This distribution differs from other SPECT reports, but is similar to that for preservation observed with structural magnetic resonance imaging (MRI). This suggests that SPECT preservation may arise from age related changes in brain anatomy, not regional cerebral blood flow (rCBF), and we demonstrate that it can arise from the partial-volume effect in areas where white matter contracts with age. Age related losses extended over the whole pre-frontal midline area and an extended pattern of focal losses was seen in the peripheral cortex that was consistent with major sulci. There were also focal losses in the cerebellum. The most significant SPECT loss was in the anterior cingulate, although no structural changes were observed there in the MRI study. A model of sulcal widening at the junction of the inter-hemispheric fissure and cingulate sulcus, when degraded by the partial-volume effect, could explain this anterior cingulate loss. CONCLUSION: Optimized processing has revealed spatial patterns for age related preservation and losses in brain SPECT that indicate their origin is primarily structural. Correction for structural effects in optimized SPECT is needed to confirm whether any regional ageing effects derive from changes in rCBF. PMID- 15891593 TI - Scintigraphic prediction of left ventricular functional recovery early after primary coronary angioplasty using single-injection quantitative electrocardiographic gated SPECT. AB - OBJECTIVE: The clinical usefulness of characterizing reperfused myocardium by perfusion/thickening assessment using electrocardiographic gated single photon emission computed tomography (SPECT) has not been investigated. We evaluated whether single-injection gated SPECT with 99mTc tetrofosmin early after primary percutaneous coronary intervention (PCI) can predict left ventricular (LV) functional recovery. METHODS: Gated SPECT was performed 3 days after primary PCI in 45 patients with acute myocardial infarction and revascularized segments were classified into perfusion/thickening mismatched segments, matched normal and matched abnormal segments. Gated SPECT was repeated 3 months later to evaluate the changes in LV ejection fraction (deltaLVEF). RESULTS: Among 332 revascularized segments, there were 83 mismatched segments, 163 matched abnormal segments and 86 matched normal segments. In all the patients, LVEF increased significantly from 3 days to 3 months after primary PCI (52+/-13 to 57+/-14%, P<0.0001). Patients were divided into two groups according to deltaLVEF: 24 patients with LV functional recovery (deltaLVEF > or = 5%) and 21 patients without LV functional recovery. The number of mismatched segments in patients with LV functional recovery was significantly greater than that in patients without (2.7+/-1.7 vs. 0.8+/-1.4, P<0.0003) despite no differences in the number of matched abnormal and matched normal segments. There was a significant correlation between deltaLVEF and the number of mismatched segments (r=0.56, P<0.0001) and LVEF at 3 months after primary PCI was related to the number of matched abnormal segments (r=-0.78, P<0.0001). CONCLUSION: Single-injection gated SPECT early after primary PCI can predict LV functional recovery. PMID- 15891594 TI - Comparison of cardiac to hepatic uptake of 99mTc-tetrofosmin with and without adenosine infusion to predict the presence of haemodynamically significant coronary artery disease. AB - BACKGROUND: Coronary artery disease impairs cardiac vasodilatory reserve. A low ratio of cardiac to hepatic vasodilatory reserve may be diagnostic for coronary artery disease. AIM: To compare the ratio of cardiac to hepatic uptake of 99mTc tetrofosmin during adenosine infusion and at rest in patients with and without coronary artery disease in order to determine whether the ratio was significantly different between the two groups. METHODS: Fifty-one patients who underwent coronary angiography and adenosine stress myocardial perfusion imaging using 99mTc-tetrofosmin were studied retrospectively. Anterior planar images from the single photon emission computed tomography (SPECT) raw data were used to draw regions of interest around the heart and liver. The counts per pixel in each region were used to calculate the stress ratio (SR) and the rest ratio (RR) as follows: SR = (cardiac counts per pixel)at stress/(hepatic counts per pixel)at stress; RR = (cardiac counts per pixel)at rest/(hepatic counts per pixel)at rest. The SR and SR/RR ratios were compared in patients with and without significant coronary artery disease. Receiver operating characteristic curves were drawn for SR and SR/RR. RESULTS: The SR and SR/RR ratios were significantly lower in patients with significant coronary artery disease than in patients without (P<0.001). A cut-off ratio of SR/RR = 1.00 yielded 87% sensitivity and 74% specificity for the detection of significant coronary artery disease. Combining SR/RR with standard SPECT image interpretation increased the sensitivity without substantially changing the specificity in comparison with standard SPECT image interpretation only. CONCLUSION: Comparison of cardiac to hepatic 99mTc tetrofosmin concentration at rest and under adenosine stress provides useful diagnostic information for the assessment of the presence of significant coronary artery disease. PMID- 15891595 TI - Reliability of one-point blood sampling method for calculating input function in Na18F PET. AB - OBJECTIVE: Conventional methods of quantitative Na18F positron emission tomography require multiple arterial blood sampling in order to obtain the input function, and the procedures are invasive and complicated. This study aims to establish a simplified and reliable technique for obtaining the input function. METHODS: Multiple arterial blood sampling was performed on 12 persons. The time point for one-point sampling was determined as the time when (1) the plasma radioactivity obtained showed the highest correlation to the real integrated value, which was calculated from the input function, and (2) the coefficient of variation of the real integrated value divided by plasma radioactivity obtained at each time point became the minimum. Scaling factors were obtained in order to estimate the plasma radioactivity at each time point, and a reference table was produced in order to estimate the input function. RESULTS: The optimal timing for one-point sampling was 12 min after intravenous injection of Na18F. The estimated integrated value obtained from arterial blood sampling at 12 min and the reference table was highly correlated with the real integrated value (P<0.001). Levels of plasma radioactivity of arterial blood and venous blood were almost the same at 12 and 40 min after Na18F injection. Percentage errors in the estimation of the integrated value were 2.63% (n=12) for the arterial blood collected at 12 min and 4.14% (n=12) for the venous blood collected at 30 min. CONCLUSIONS: This simplified method is clinically applicable and would replace traditional methods that require multiple blood sampling. PMID- 15891596 TI - Prokinetic therapy: what can be measured by gastric scintigraphy? AB - OBJECTIVE: To evaluate whether gastric scintigraphy with quantitative analysis of gastric peristalsis may be a useful tool for documenting the effects of prokinetic therapy. METHODS: Gastric emptying was determined in eight patients with insulin dependent diabetes mellitus (IDDM) and nine patients with progressive systemic sclerosis (PSS) after ingestion of a semi-solid test meal. Fourier analysis of condensed images was used to evaluate contraction amplitudes of the entire stomach, as well as frequency and velocity of gastric contractions. Patients were studied twice, first under baseline conditions and then after 4 weeks of continuous oral prokinetic therapy with cisapride (10 mg t.i.d.). Findings were compared with data derived from 15 healthy controls. RESULTS: At baseline conditions IDDM and PSS patients exhibited reduced gastric peristalsis and delayed emptying compared to controls. After 4 weeks of prokinetic treatment, gastric contraction amplitudes increased significantly, especially in the gastric corpus and the proximal stomach, associated with a slight improvement of gastric emptying. Changes in frequency and velocity of gastric contractions during prokinetic treatment correlated with the improvement of gastric motility and emptying. CONCLUSIONS: Prokinetic therapy with cisapride stimulates gastric motility, and could be non-invasively monitored by scintigraphy. This radionuclide technique provides a number of relevant parameters of gastric function, such as gastric emptying, contraction amplitudes, frequency and velocity of gastric contractions, which give crucial insights into the mechanisms of action of prokinetic drugs. PMID- 15891597 TI - Pharmacokinetics and normal scintigraphic appearance of 99mTc aprotinin. AB - OBJECTIVE: To confirm the pharmacokinetics and biodistribution of 99mTc aprotinin in normal volunteers and to determine the optimum time for scanning post injection, prior to further investigations of 99mTc aprotinin as an imaging agent for amyloidosis. METHODS: Five patients (three men and two women, average age 49 years, age range 38-66 years) without a history of amyloidosis or any of the associated diseases, were included in this prospective study. Blood and urine were collected and images were performed of the whole body and wrists. CONCLUSIONS: Normal biodistribution of 99mTc aprotinin includes early cardiac and lung activity in the blood pool phase with subsequent hepatic activity and renal excretion with variable splenic activity. There is variable bowel uptake on later images. The best quality images were obtained 90 min post-intravenous administration, and this is likely to be the optimum time for clinical imaging. PMID- 15891598 TI - Impact of waiting on the perception of service quality in nuclear medicine. AB - BACKGROUND: This is the first study examining the link between waiting and various dimensions of perceived service quality in nuclear medicine. METHODS: We tested the impact of selected waiting experience variables on the evaluation of service quality, measured using the Servqual tool, of 406 patients in nuclear medicine, with objective and subjective waiting times as co-variates. The sequence of events in service delivery in nuclear medicine (waiting time before injection, waiting time before scanning and total waiting time) is taken into account. RESULTS: Patients underestimated the waiting time before injection and the total waiting time, while overestimated the waiting time before scanning. Our results show that the total subjective waiting time has more impact on the reliability dimension (R2(adj)=0.148) than on the other service quality dimensions of Servqual. Providing information about the reasons for delay had a significant main effect on the perception of reliability (F=9.64, P=0.02). CONCLUSIONS: The study stresses the importance of explaining the causes for delay to increase patients' perceptions of reliability of the nuclear medicine department and contains several findings that should assist service managers to formulate more effective waiting perception strategies. PMID- 15891599 TI - 99mTc demotate 1: biodistribution and elimination characteristics in rats. AB - BACKGROUND AND METHODS: In this study, we investigated the biodistribution and elimination characteristics of a new radiolabelled somatostatin analogue, 99mTc demotate 1, in rats by in-vivo biodistribution and elimination experiments, perfused rat liver and kidney experiments and micro-autoradiography of renal tissue. RESULTS: Rapid clearance from blood and most organs was found. High and long-term uptake in organs with high density of somatostatin receptors (the adrenals and pancreas) and in stomach and intestine was reduced in non radiolabelled octreotide pretreated animals. The predominant urine excretion was associated with an accumulation of 99mTc demotate 1 in the kidney, mainly in the renal cortex. This uptake was not affected by non-radiolabelled octreotide pretreatment. CONCLUSION: 99mTc demotate 1 is a prospective radiopharmaceutical for use in human medicine in somatostatin receptor-positive tumour imaging and its potential should be confirmed in further experiments and clinical trials. PMID- 15891600 TI - Long-lived radiopharmaceuticals: dispensing from multidose vials. AB - We report the use of broth simulation as a means of validating the practice of sub-dispensing from stock vials of long-lived sterile radiopharmaceuticals. 'Matched' vials of nutrient broth accompanied the stock vials of long-lived sterile radiopharmaceuticals during their time in use and were subject to the same handling and storage procedures. At the end of the life of each radiopharmaceutical stock vial, the matching broth residue was sent for incubation and reporting. The results to date have yielded no reports of microbial contamination in any of the simulated broths (0 in 256). PMID- 15891603 TI - Can you prevent ventilator-associated pneumonia? PMID- 15891605 TI - Trauma care. PMID- 15891606 TI - Cardiac care. PMID- 15891607 TI - Respiratory care. PMID- 15891609 TI - Outmuscling rhabdomyolysis. PMID- 15891611 TI - NP educators in clinical practice. PMID- 15891612 TI - Health problems and mental illness. PMID- 15891613 TI - Preventing obesity in schools. PMID- 15891614 TI - Diabetes update: 2005 ACE Guidelines. PMID- 15891615 TI - Many factors needed to solve malpractice woes. PMID- 15891616 TI - ARVD: a silent killer. PMID- 15891618 TI - Viral or acute bacterial rhinosinusitis? Determining the difference. PMID- 15891620 TI - Rotator cuff versus cervical spine: making the diagnosis. PMID- 15891621 TI - Menactra for prevention of meningococcal disease. PMID- 15891622 TI - Learn the business skills of advanced practice nursing. PMID- 15891623 TI - Terminating the provider-patient relationship. PMID- 15891628 TI - Consortium of otolaryngology-head and neck surgery journals to collaborate in maintenance of high ethical standards. PMID- 15891629 TI - The effects of basic fibroblast growth factor on postoperative mastoid cavity problems. AB - OBJECTIVES: To present the effects of basic fibroblast growth factor on intractable cavity problems and discuss the mechanisms of its effects. METHODS: We treated three ears with postoperative open cavities. All three cases had suffered from chronic discharge of the ear for 7, 10, and 30 years, respectively; 100 microg/ml of trafermin (genetic recombination) solution, as basic fibroblast growth factor, was dropped into the open cavity once daily. If bacterial and/or fungal infection was observed, antibiotics and/or antifungal agents were administered locally twice daily. RESULTS: The cavities epithelialized and were cured within 2 months using this treatment. CONCLUSION: Our results suggest that basic fibroblast growth factor stimulates the proliferation and differentiation of keratinocytes, fibroblasts, and endothelial cells, leading to accelerated wound healing. The basic fibroblast growth factor agent appears to be highly effective in treating intractable cavity problems. PMID- 15891630 TI - Pars flaccida displacement pattern in otitis media with effusion in the gerbil. AB - HYPOTHESIS: Our hypothesis is that otitis media with effusion causes stiffness loss in the pars flaccida of the tympanic membrane. This loss of stiffness may be persistent and may trigger the development of retraction pockets and cholesteatoma. BACKGROUND: Otitis media with effusion is a very common disease in childhood. It can cause minor to moderate hearing loss and delayed speech development. Otitis media with effusion is a risk factor for retraction pocket formation. METHODS: Otitis media with effusion was induced unilaterally in 15 gerbils by obstructing the eustachian tube with glue. The contralateral ears served as normal controls. As a measure of mechanical stiffness, pars flaccida peak displacement versus pressure was used. The displacement measurements were made with moire interferometry. This is a noncontacting optical technique with which the displacement of an object can be measured in real time. RESULTS: The mean peak displacement of the pars flaccida in the group with otitis media with effusion was increased threefold as compared with normal controls. This difference was statistically significant. CONCLUSION: There was a loss of mechanical stiffness in the pars flaccida caused by otitis media with effusion. This loss of stiffness may be persistent and may predispose for retraction pocket formation and cholesteatoma development. PMID- 15891631 TI - Early and late effects of capsaicin pretreatment in otitis media with effusion. AB - PURPOSE: Discovery of the role of the neurogenic inflammation in the formation of otitis media with effusion has led to the investigation of the place of some neuropeptide antagonists in the treatment. In the current study, we investigated the effect of capsaicin (CP) pretreatment on the inflammation and proliferation in the middle ear mucosa and on the nerve fibers containing substance P, vasoactive intestinal polypeptide, and calcitonin gene-related peptide. METHODS: Seventeen Wistar rats were used in the study. Ten rats were given CP on 3 consecutive days, and seven rats were given isotonic saline solution. Seven days after the third injection, animals were operated on, and their eustachian tubes were occluded. On the seventh day after the operation, five rats from the test group and three from the control group were killed. The others were killed 21 days after the operation. In the histopathologic examination of the sections, acute inflammation and proliferation scores were determined. Gland degeneration, goblet cell hyperplasia, and the density of mast cells were evaluated. Neural elements were stained immunohistochemically. RESULTS: The acute inflammation score in the test group was lower, but the difference was insignificant (p > 0.05). The proliferation score in the test group was lower, and the difference was significant (p = 0.02). In the control group, gland degeneration was significantly higher (p = 0.044). Goblet cell hyperplasia demonstrated no difference between two groups (p > 0.05). Mast cell density was higher in the control group, but the difference was not significant (p > 0.05). Substance P immunoreactivity (IR) was significantly higher in the control group (p = 0.015). calcitonin gene-related peptide-IR and vasoactive intestinal polypeptide-IR were limited in both groups. CONCLUSIONS: That CP pretreatment reduces inflammatory proliferative findings, and gland degeneration leads us to consider that it could be effective in both treatment of experimental otitis media with effusion and prevention of its complications. PMID- 15891632 TI - Effects of topical application of mitomycin-C and 5-fluorouracil on myringotomy in rats. AB - OBJECTIVE: The effects of topical application of Mitomycin-C (MMC) and 5 fluorouracil (5-FU) for maintaining myringotomy patency were investigated in this experimental study. STUDY DESIGN: We performed simple myringotomy with a knife on 140 tympanic membranes of 70 rats. Rats were divided in two study groups and a control group. Each study group had 60 tympanic membranes, and the control group had 20. We applied Mitomycin-C (0.4 mg/ml) in Group A, 5-fluorouracil (50 mg/ml) in Group B topically, and sterile saline in the control group for 10 minutes. Examination was made with otoendoscope on days 1, 3, 5, 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, and 70, and patency rates were recorded. RESULTS: MMC and 5-FU Groups remained open for a mean of 46.17 days and 14.62 days, respectively. The control ears healed within 10.4 days. Fibrosis of the MMC treated group was the same as that of 5-FU-treated groups. Fibrosis of both study groups was significantly lower than that of the control group. CONCLUSIONS: MMC is more effective than 5-FU, which is more effective than the simple myringotomy procedure in extending the patency of myringotomies in rat tympanic membranes (p < 0.05). Both medications are useful as an adjunct in preventing myringotomy closure. PMID- 15891633 TI - The effect of radiofrequency and mitomycin C on the closure rate of human tympanostomy. AB - OBJECTIVES: To conduct a prospective, randomized, controlled trial determining the feasibility of radiofrequency with or without topical mitomycin C application in delaying the closure time of human tympanostomy and screening its efficacy in management of recurrent acute otitis media and otitis media with effusion. METHODS: From November 2002 to January 2004, 96 patients (180 ears) who were to undergo surgical intervention for recurrent acute otitis media or otitis media with effusion were included in this study. Sixty ears with a diagnosis of recurrent acute otitis media were equally randomized to three procedures: cold knife myringotomy (Group A), radiofrequency tympanostomy (Group B), and radiofrequency tympanostomy with topical mitomycin C application (Group C). One hundred twenty ears diagnosed as having otitis media with effusion were equally randomized to the last two procedures only. Patients were followed-up every week in the first month, every 2 weeks in the second month, and monthly after that until closure of the tympanostomy. A special follow-up setting was designed 3 months after tympanostomy closure where tympanograms and pure-tone audiograms were obtained. RESULTS: Groups B and C showed a significantly slower rate of closure than Group A. At the same time, Group C demonstrated a slower rate when compared with Group B. The mean closure time of Group B was 3.5 weeks, with no difference between recurrent acute otitis media and otitis media with effusion. In contrast, the mean closure times for Group C were 5.3 and 7 weeks in cases of otitis media with effusion and recurrent acute otitis media, respectively, with the latter demonstrating a significantly slower closure rate. No recurrence of acute otitis media was reported in 10, 45, and 80% of Groups A, B, and C, respectively. Fifty-nine percent of Group C showed no clinical or tympanometric signs of otitis media with effusion 3 months after closure of tympanostomy. In contrast, Group B reported a lower cure rate (28%), with a statistically significant difference between both groups. Absence of recurrence and improvement in tympanometry signs correlated significantly with the duration of patency of tympanostomy (p < 0.01). The success rate was higher in patients who underwent adenoidectomy, but this did not reach statistical significance except in Group C (p < 0.01). CONCLUSION: Radiofrequency and mitomycin C delay the closure rate of human tympanostomy. The efficacy of mitomycin C seems to be amplified in the presence of an inflamed tympanic membrane. Radiofrequency-assisted mitomycin enhanced tympanostomy is a precise, safe, and cost-effective procedure that provides a viable approach in the management of recurrent acute otitis media and otitis media with effusion, especially when used in conjunction with adenoidectomy. PMID- 15891634 TI - Implications of laser assisted tympanostomy in adults. AB - OBJECTIVES: To assess outcome in adult individuals undergoing laser-assisted tympanostomy without ventilation tube placement. STUDY DESIGN: Case series with 2 month follow-up. SETTING: Faculty practice, research protocol, tertiary care academic medical center. PATIENTS AND METHOD: Laser-assisted tympanostomy was performed on a total of 142 ears (108 individuals). Indications included serous otitis media with effusion (66 ears/47 patients), functional eustachian tube dysfunction (48 ears/36 patients), acute otitis media (19 ears/16 patients), and endoscopic visualization of the middle ear (9 ears/9 patients). INTERVENTION: The laser-assisted tympanostomy procedure is performed with a CO2 laser under local anesthesia on an outpatient basis. MAIN OUTCOME MEASURES: Patency time of the tympanostomy, presence of fluid after the closure of the tympanostomy, tympanometry and tone audiometry findings, relief of symptoms. RESULTS: Middle ear disease was resolved after the closure of tympanostomy in 47.9% of patients with serous otitis media with effusion. In 79.1% of patients with functional eustachian tube dysfunction, symptoms were diminished. All patients with acute otitis media had a satisfactory outcome. Laser-assisted tympanostomy was found to be quite helpful in patients undergoing middle ear endoscopy. CONCLUSIONS: Laser assisted tympanostomy without ventilation tubes provides a safe alternative surgical option in adult patients in certain cases. The selection criteria for this procedure are addressed in detail. PMID- 15891635 TI - Bone cement ossiculoplasty: incus to stapes versus malleus to stapes cement bridge. AB - OBJECTIVE: In this study, our purpose was to evaluate results of our experience with bone cement repair of ossicular discontinuity between the incus and stapes and between the malleus and stapes. METHODS: Medical records of patients who underwent surgery for chronic otitis media between March 2000 and December 2002 were evaluated retrospectively. Fifty-seven patients who underwent bone cement ossiculoplasty and had appropriate follow-up data were included in the study. Bone cement reconstruction of the ossicular chain was performed 1) from incus to stapes (I-S) in the absence of long arm or lenticular process of the incus and 2) from malleus to stapes (M-S) in the absence of the incus. The clinical data of the patients were evaluated by otoscopic examination and audiometry. RESULTS: The graft take rate was 84.1%. I-S procedure was performed in 42 and M-S in 8 patients. Pre- and postoperative PTAs of all patients were compared, which showed a significant improvement in air PTA (p < 0.001) while bone PTA did not change (p > 0.05). In I-S and M-S groups, successful hearing restoration could be achieved in 78.6.1% and 87.5% of the patients, respectively. Hearing results of different aural pathologies (chronic otitis media and conductive hearing loss) and surgeries (tympanotomy and tympanoplasty with or without mastoidectomy) were not significantly different (p > 0.05). CONCLUSION: Bone cement ossiculoplasty offers cost effective and significant improvement in conductive hearing loss. PMID- 15891636 TI - Malleus-to-footplate ossicular reconstruction prosthesis positioning: cochleovestibular pressure optimization. AB - AIMS: To determine 1) the best position for hydroxylapatite malleus-to-footplate (MFP), ossicular replacement prosthesis (ORP) in reconstructed ears, and 2) whether preserving the stapes superstructure (SS), when present, has acoustic advantages. BACKGROUND: Positioning of the MFP-ORP head beneath the neck of the malleus may produce maximal force, whereas positioning beneath the manubrium of the malleus may produce the greatest displacement. It is not clear which is the optimal placement position. In addition, we look at the effect of the SS on sound transmission to the inner ear in ossicular reconstruction. METHODS: The ear-canal air pressure and vestibular hydro-pressure were measured in human cadaver temporal bones with incus intact, removed, and replaced with the MFP-ORP; the ORP head was placed at three different positions on the malleus (head, mid-manubrium, and umbo) while keeping its base at the center of stapes footplate with intact or removed stapes SS. The vestibular pressure ratio between the ear with intact incus and MFP-ORP reconstructed ear is defined as Lmfp, the loss caused by the prosthesis in relation to the normal ossicular chain. RESULTS: The mean magnitude of Lmfp, averaged in the important speech frequency region of 0.5 to 3 kHz, is approximately 7.8 dB at the neck with stapes SS. In comparison, mean magnitude of Lmfp for mid-manubrium without stapes SS is 15 dB (p = 0.04), and with the stapes SS it is 16 dB (p = 0.05), whereas at the umbo without SS it is 15 dB (p = 0.03). In the 8 kHz region, the mean magnitude of Lmfp is approximately 1 dB with the stapes SS intact and approximately 8.5 dB when it was removed (p < 0.09). CONCLUSION: There are significant physiologic advantages to placing the hydroxylapatite MFP-ORP beneath the neck of the malleus and preserving the SS. PMID- 15891637 TI - Bacterial adherence to titanium surface coated with human serum albumin. AB - HYPOTHESIS: An albumin coating on titanium implants will inhibit bacterial adhesion on the implant surface. BACKGROUND: Bacterial, protein, and platelet adhesion on otologic implants and tympanostomy tubes is a major reason for implant sequelae and can eventually lead to implant removal. The role of albumin coating of the implant in prevention of protein adhesion on implant surface has already been tested by the authors. In the present study the authors examined the in vitro adherence of Staphylococcus aureus and Pseudomonas aeruginosa on an albumin-coated and uncoated titanium surface. METHODS: Human serum albumin (HSA) coated and uncoated titanium surfaces were exposed to viable S. aureus and P. aeruginosa and, after washings, photographed by fluorescence microscopy to quantify the adhered bacteria, which was stained with acridine orange. RESULTS: Bacteria in the suspension adhered at a significantly lesser rate to the coated surfaces than to the uncoated surfaces, with overall bacterial adhesion dependent on bacterial concentration. Binding of S. aureus on HSA-coated surfaces was inhibited significantly (from 82 to 95% depending on concentration). Binding of P. aeruginosa was inhibited from 29 to 37%. CONCLUSION: Because albumin coating can reduce bacterial adherence on titanium surfaces in vitro, reduction is possible in bacterial contamination and infection of the HSA-coated titanium implant in vivo. PMID- 15891638 TI - Study of the posterior communicating arteries of the circle of willis in idiopathic sudden sensorineural hearing loss. AB - OBJECTIVES: To evaluate the incidence of non-functioning posterior communicating arteries (PCoA) of the circle of Willis in patients presenting with idiopathic sudden sensorineural hearing loss (SSHL). BACKGROUND: Pathogenesis of SSHL remains unclear despite several theories. A strong association between non functioning PCoA of the circle of Willis and SSHL has been recently suggested, supporting the theory of vascular disorders. SETTING: Tertiary referral center. PATIENTS AND METHODS: Ultrasonographic Doppler flow measurements of the vertebrobasilar system were performed prospectively in 27 consecutive patients with idiopathic SSHL and in 27 healthy subjects. RESULTS: A non-functioning PCoA was found in 4 patients with idiopathic SSHL and in 4 cases in the control group. CONCLUSION: The results of our study did not show any association between non functioning PCoA of the circle of Willis and idiopathic SSHL, contrasting with previous data. We do not recommend adding routine transcranial Doppler ultrasonography in the test-battery for SSHL. PMID- 15891639 TI - Paget disease and sensorineural hearing loss associated with spiral ligament degeneration. AB - HYPOTHESIS: Previously unreported cystic degeneration of the spiral ligament in cases of Paget disease. BACKGROUND: About 70% of cases of Paget disease involve the skull, with hearing affected in approximately 50% of these. The hearing impairment may be sensorineural, mixed, or, rarely, only conductive. The etiology and pathogenesis of the hearing loss are not yet understood, and reports in the literature are inconsistent regarding the pathologic changes responsible for sensorineural hearing loss. Of six pairs of temporal bones from patients with Paget disease in the temporal bone collection of a research institution, two pairs have abnormalities not previously associated with sensorineural hearing loss or Paget disease. We report the histopathologic findings in these temporal bones. METHODS: The temporal bones were fixed in formalin, decalcified in ethylenediaminetetraacetic acid, embedded in celloidin, and sectioned in the horizontal plane at a thickness of 20 microm. Every 10th section was stained with hematoxylin-eosin and mounted on glass slides. The sections were examined by light microscopy. RESULTS: Cystic degeneration of the spiral ligament, primarily in the basal segment, was found in both cases. Endolymphatic hydrops and a small endolymphatic sac with calcification of the perisaccular tissue were found in one case. CONCLUSIONS: Cystic degeneration of the spiral ligament has not been previously reported and may be unique to Paget disease. This is consistent with recent literature showing a previously unsuspected role of the spiral ligament in sensorineural hearing loss. PMID- 15891640 TI - Glutathione S-transferase gene polymorphisms in presbycusis. AB - HYPOTHESIS: Glutathione and glutathione-related antioxidant enzymes are involved in the metabolism and detoxification of cytotoxic and carcinogenic compounds as well as reactive oxygen species. Reactive oxygen species generation occurs in prolonged relative hypoperfusion conditions such as in aging. The etiology of presbycusis is much less certain; however, a complex genetic cause is most likely. The effect of aging shows a wide interindividual range; we aimed to investigate whether profiles of (glutathione S-transferase (GST) M1, T1 and P1 genotypes may be associated with the risk of age-related hearing loss. PATIENTS AND METHODS: We examined 68 adults with presbycusis and 69 healthy controls. DNA was extracted from whole blood, and the GSTM1, GSTT1 and GSTP1 polymorphisms were determined using a real-time polymerase chain reaction and fluorescence resonance energy transfer with a Light-Cycler Instrument. Associations between specific genotypes and the development of presbycusis were examined by use of logistic regression analyses to calculate odds ratios and 95% confidence intervals. RESULTS: Gene polymorphisms at GSTM1, GSTT1, and GSTP1 in subjects with presbycusis were not significantly different than in the controls (p > 0.05). Also, the combinations of different GSTM1, GSTT1, and GSTP1 genotypes were not an increased risk of presbycusis (p > 0.05). CONCLUSION: We could not demonstrate any significant association between the GSTM1, GSTT1, and GSTP1 polymorphism and age-related hearing loss in this population. This may be because of our sample size, and further studies need to investigate the exact role of GST gene polymorphisms in the etiopathogenesis of the presbycusis. PMID- 15891641 TI - Inner ear abnormalities in patients with Goldenhar syndrome. AB - OBJECTIVE: The objective of this study is to investigate the inner ear malformations in patients with Goldenhar syndrome and to hypothesize the potential embryopathogenesis of these malformations. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Fourteen patients with Goldenhar syndrome. INTERVENTIONS: Each patient underwent hearing tests and high resolution computed tomography (CT) of the temporal bone. In six patients, magnetic resonance imaging of the temporal bone also was performed. RESULTS: Among the 14 patients with Goldenhar syndrome, 13 had outer and middle ear anomalies and 5 (36%) had inner ear malformations, including one case of common cavity. CONCLUSIONS: Our observations regarding inner ear anomalies in Goldenhar syndrome correlate with the reported cases in the literature and may help to hypothesize the embryological origin of these malformations, which can caused by an early developmental arrest in the fourth gestational week. Specialists evaluating patients with Goldenhar syndrome should be aware of the possibility of inner ear malformations, which could be diagnosed earlier with appropriate imaging studies. PMID- 15891642 TI - Sensorineural hearing impairment in patients with Pmp22 duplication, deletion, and frameshift mutations. AB - OBJECTIVE: To characterize and distinguish the types of sensorineural hearing impairment (SNHI) that occur in hereditary motor and sensory neuropathy Type 1a (HMSN-1a) and hereditary neuropathy with liability to pressure palsies (HNPP), which are caused by deletion or frameshift mutation. STUDY DESIGN: Prospective study. SETTING: Ambulatory patients in a university hospital. PATIENTS: Twelve patients with HMSN-1a due to a duplication of the PMP22 gene on chromosome 17p11.2, 16 patients with HNPP due to the common PMP22 deletion (HNPP del), and 11 HNPP patients with a frame shift mutation (heterozygous PMP22 G-insertion) (HNPP mut), all confirmed by molecular genetic analysis. INTERVENTIONS: Pure-tone audiograms and speech audiograms were obtained. MAIN OUTCOME MEASURES: Results of cross-sectional analysis comprising linear regression of hearing threshold on age. RESULTS: Pure-tone audiograms showed mild to moderate SNHI, predominant at the low and the high frequencies. SNHI showed significant progression by approximately 0.4 dB per year at 0.25 to 4 kHz and up to 1 to 2 dB per year at 4 to 8 kHz. Patients with HMSN-1a had substantial, presumably congenital, SNHI but did not show significant progression beyond presbyacusis. Patients with HNPP showed postnatal onset at age 11 years with progression of SNHI in excess of presbyacusis by 0.4 dB per year. All three types of neuropathy showed normal speech recognition. CONCLUSIONS: All three types of neuropathy showed SNHI with normal speech recognition. HMSN-1a showed stable SNHI without progression beyond presbyacusis. HNPP showed progression beyond presbyacusis with postnatal onset. The differences in SNHI may be explained by the differences in PMP22 expression. The progressive SNHI in HNPP might be explained by the liability for exogenous factors associated with this disorder. PMID- 15891643 TI - Inner ear abnormalities in a Kcnq1 (Kvlqt1) knockout mouse: a model of Jervell and Lange-Nielsen syndrome. AB - HYPOTHESIS: Mice lacking functional KCNQ1 (previously known as KvLQT1) channels exhibit functional and structural abnormalities that indicate disturbed production of endolymph. BACKGROUND: Congenital deafness associated with cardiac conduction abnormalities (Jervell and Lange-Nielsen syndrome) is associated with dysfunctional KCNQ1/KCNE1 channel complex. This potassium channel plays a critical role in the production and homeostasis of endolymph by the stria vascularis. A preliminary report documented severe abnormalities of the scala media and vestibular compartments of a single mouse lacking functional KCNQ1 alleles. METHODS: Hearing thresholds were measured in three Kcnq1 knockout mice, two heterozygous mice, and one wild-type mouse by auditory brainstem response recordings using clicks, after which the temporal bones were removed. After fixation and dehydration, the ears were embedded in araldite, sectioned at 20 microm thickness, stained with toluidine blue on glass slides, and examined with the light microscope. RESULTS: Kcnq1 knockout mice were deaf and demonstrated circling behavior. They exhibited a marked atrophy of the stria vascularis, contraction of the endolymphatic compartments, and collapse and adhesion of surrounding membranes. There was a complete degeneration of the organ of Corti and an associated degeneration of the spiral ganglion. CONCLUSION: Kcnq1 knockout mice exhibit histopathologic findings that are comparable to those reported in human temporal bone cases of Jervell and Lange-Nielsen syndrome, and provide further evidence that KCNQ1 channel dysfunction can lead to congenital deafness in this syndrome. PMID- 15891644 TI - Neurostimulation as a new treatment for severe tinnitus: a pilot study. AB - BACKGROUND: Tinnitus is an uncomfortable symptom for the patient and an embarrassing one for the consulted physician. So far, there is no treatment that can be considered well established in terms of providing long-term reduction of tinnitus in excess of placebo effects. There is considerable evidence of pathophysiological similarity between tinnitus and chronic pain. Some forms of chronic pain can be treated by neurostimulation. OBJECTIVE: This study was designed to investigate the feasibility of neurostimulation of the cochlear nerve in order to reduce tinnitus. STUDY DESIGN: Pilot study. SETTING: Tertiary referral center. PATIENTS: Five patients with therapeutically refractory tinnitus were selected for this study. INTERVENTION: Placing a stimulation lead around the cochlear nerve through the suboccipital approach and connecting the stimulation lead to a pulse generator. MAIN OUTCOME MEASURES: The patients experienced 1) an absence of major or minor complications, such as death, meningitis, cranial nerve deficit, and vestibular problems; 2) tolerance of the procedure as considered by the patient; 3) relief of tinnitus in at least one patient. RESULTS: Implantation of the neurostimulation system was accomplished in each patient without any difficulty. None of the patients considered the treatment unbearable. No major or minor complications occurred in this study. Subjective tinnitus reduction was accomplished in four patients. CONCLUSION: Our preliminary data show that neurostimulation of the cochlear nerve is feasible, is bearable for the patient, and is a safe treatment modality without major complications. The effects on tinnitus are promising. PMID- 15891645 TI - Sound localization and sensitivity to interaural cues in bilateral users of the Med-El Combi 40/40+cochlear implant system. AB - OBJECTIVE: The purpose of this study was to investigate sound localization in subjects bilaterally implanted with MED-EL COMBI 40/40+ cochlear implants. In addition, the sensitivity to interaural cues was assessed. METHODS: In the localization test (11 subjects), CCITT noise (500 ms, original and HRTF-filtered, 70/75/80 dB sound pressure level) was presented from one of seven loudspeakers between -90 degrees and 90 degrees azimuth. The subject had to indicate which loudspeaker the noise was presented from. Sensitivity to interaural level differences (ILD) was assessed by performing localization tests (4 subjects) with the loudness of the two speech processors unbalanced to various degrees. To investigate the subjects' sensitivity to interaural time differences (ITD), lateralization was measured (7 subjects) as a function of the time difference between two Gaussian-like pulses, each directed to one of the subject's speech processor microphones by way of headphones. RESULTS: The judgments of all subjects significantly correlated with the positions of the loudspeakers. The scatter in the judged azimuth measured by the standard deviation of the responses was on average 27.5 degrees . Unbalanced loudness of the speech processors produced a bias in azimuth toward the speech processor with the louder volume setting. The mean rate of shift was 1.4 degrees per unit on the Wuerzburg loudness scale. Six of seven subjects showed a significant sensitivity to ITDs with the approximate time difference required for complete lateralization being 1,200 micros on an average. The one subject not showing a sensitivity to ITDs performed worst in the localization test. CONCLUSIONS: Bilateral cochlear implantation can restore spatial hearing in cochlear implant users. Both ILDs and ITDs are used by bilateral cochlear implant users in sound localization with ILDs appearing to be the dominant cue. PMID- 15891646 TI - Ventilation tubes and cochlear implants: what do we do? AB - OBJECTIVE: To determine current practice management with respect to ventilation tubes and cochlear implants. STUDY DESIGN: Questionnaire. SETTING: All members of the American Neurotology Society were sent questionnaires. MAIN OUTCOME MEASURES: Members were asked a series of questions including how they deal with ventilation tubes before cochlear implantation, how they manage serous otitis media in patients undergoing cochlear implantation, and how they manage otitis prone children with cochlear implants. RESULTS: Two hundred and twenty members returned questionnaires. Surgeons who replied perform an average of 25 implants per year: 15 in adults and 10 in children. Analysis of the data revealed a wide practice variation between surgeons. Fifty-six percent of surgeons will place a cochlear implant in a patient with a clean, dry ventilation tube in place. More than half the surgeons will place a ventilation tube in a child with serous otitis media, let the ear settle down, and perform the cochlear implant at a second operation. Wide variation in the management of otitis prone children with cochlear implants exists with respect to placement of ventilation tubes. Only 5% of surgeons reported any complications with cochlear implants that they attributed to ventilation tubes. There were a number who suggested their practice had changed since the recent identification of issues involving meningitis in implantees. CONCLUSION: Wide practice variation exists with the management of ventilation tubes in cochlear implant patients. On the basis of the results of this survey, it is acceptable to place cochlear implants in patients with clean, dry ventilation tubes. It also acceptable to place ventilation tubes in otitis prone children with cochlear implants. Despite theoretic concerns, the reported incidence of complications is low. PMID- 15891647 TI - Pattern of hearing loss in a rat model of cochlear implantation trauma. AB - HYPOTHESIS: Trauma caused by cochlear implant electrode insertion is attributable to the combination of direct physical trauma and the delayed cell death of oxidative stress-injured auditory sensory cells. BACKGROUND: Histologic evaluation of cochlear implant electrode trauma has demonstrated that the extent of sensory cell losses is proportional to the degree of injury. However, the impact of delayed oxidative stress within injured cochlear tissues and the progressive loss of injured hair cells by way of apoptosis are at present unknown. METHODS: Laboratory rats were evaluated for hearing acuity before and after electrode insertion, before and after round window membrane incision only. Hearing was measured before trauma or incision and over the next 7 days. Objective measurements of hearing function were distortion products of otoacoustic emissions (DPOAEs) in the frequency range of 2 to 32 kHz and tone burst (i.e., 4-32 kHz) evoked auditory brain stem responses (ABRs). RESULTS: For the experimental cochleae, there were progressive increases in ABR thresholds and decreases in ABR amplitudes. The amplitude of the DPOAEs in the experimental cochleae also showed progressive decreases. For the contralateral control and round window membrane surgical control ears, there were no significant changes in either DPOAE or ABR thresholds. CONCLUSION: These results document a progressive loss of hearing acuity postimplantation and strongly suggest that electrode insertion trauma generated oxidative stress within injured cochlear tissues. PMID- 15891648 TI - An "endosteal electrode" for cochlear implantation in cases with residual hearing? Feasibility study: preliminary temporal bone experiments. AB - BACKGROUND: Over the years, an increasing number of patients with some degree of residual hearing have received cochlear implants. In these cases, the marginal benefit provided by hearing aids alone is not sufficient; however, as experience has already shown, when hearing aids are used in combination with a cochlear implant, more benefit may be obtained. As a prerequisite, this requires that residual hair cell function must remain intact postoperatively. One of the European pioneers of cochlear implants, Ernst Lehnhardt, questioned whether residual hearing might better be preserved if the implanted electrode permits the fluid-filled inner ear space to remain intact. Subsequently, he proposed insertion of a very flat electrode array design into the extraluminal space between the spiral ligament and the bony cochlear wall (endosteum). OBJECTIVE: Our study aimed to determine whether it is feasible to insert an endosteal electrode model intracochlearly but extraluminarily, anatomically, and ultimately surgically and to determine the impact on surrounding intracochlear structures. METHODS: Insertion of two silicon models of an endosteal electrode were carried out in 15 human temporal bones. Histologic examination of the temporal bones after electrode insertion was performed on both fresh and fixed specimens to determine whether the desired anatomic site of insertion was achieved. In combination with light reflected and electron microscopic techniques, the extent to which the surrounding structures were impacted was also examined. RESULTS: Successful insertion of the prototype silicon endosteal electrodes was performed intracochlearly and extraluminarly in 11 of the 15 temporal bone specimens, confirming the anatomic feasibility of insertion into the crevice between the spiral ligament and endosteum. CONCLUSIONS: On the basis of the anatomy of the human temporal bone, insertion of an "endosteal electrode" is feasible. Subsequently, in vivo animal studies are needed to determine the physical effects of insertion of an endosteal electrode design prototype upon the functionality of the surrounding intracochlear structures and in particularly the ability to preserve hearing function. PMID- 15891649 TI - Comparison of continuous interleaved sampling and simultaneous analog stimulation speech processing strategies in newly implanted adults with a Clarion 1.2 cochlear implant. AB - OBJECTIVE: This study consisted of a within-subjects comparison of speech recognition and patient preference when subjects used two different cochlear implant speech processing strategies with a Clarion 1.2 (enhanced bipolar) device: Simultaneous Analog Stimulation (SAS), and Continuous Interleaved Sampling (CIS). These two strategies used two different electrode configurations: the SAS strategy used bipolar stimulation, whereas the CIS strategy used monopolar stimulation. STUDY DESIGN: This was a multicenter study that used a within-subjects balanced crossover design. Experience with the two strategies was replicated in each subject using an ABAB design. Order of strategy use was balanced across all subjects. SETTING: The study was carried out at several cochlear implant centers affiliated with tertiary medical centers. PATIENTS: Subjects consisted of 25 postlingually deafened adults who received a Clarion cochlear implant. INTERVENTIONS: Total involvement by each subject was 14 weeks. Speech perception testing and sound quality assessments were performed after use with each strategy. MAIN OUTCOME MEASURES: Primary outcome measures include speech perception data and patient responses to questionnaires regarding speech and sound quality. RESULTS: Analyses revealed that performance did not differ significantly by the strategy encountered first as relative to the strategy encountered second and that the order in which a strategy was used did not appear to affect subjects' eventual preference for a particular strategy. Although speech recognition scores tended to be higher for CIS for most of the test measures at most of the test intervals, the analysis of variance to evaluate differences in strategy did not reveal a significant effect of strategy. Further analysis of scores obtained at the replication interval, however, revealed that scores obtained with CIS were significantly higher than scores obtained with SAS on the Hearing in Noise Test sentences in quiet and noise. In addition, significantly more patients indicated a final preference for the CIS strategy than for the SAS strategy. Importantly, both the analysis evaluating order and the analysis evaluating strategy revealed significant effects of evaluation period, indicating that time/experience with the implant had a significant effect on scores for each strategy, regardless of the order in which it was used (first or second). CONCLUSIONS: This study demonstrates that important learning occurs during the first several weeks of cochlear implant use, making it difficult to adequately compare performance with different speech processing strategies. However, the finding that patients often prefer the strategy they understand speech the best with supports the clinical practice of letting adult patients select their preferred strategy without formally evaluating speech perception with each available strategy. PMID- 15891650 TI - Pediatric cholesteatoma: canal wall window alternative to canal wall down mastoidectomy. AB - OBJECTIVE: A previous pilot series described a hybrid mastoidectomy technique, canal wall window (CWW), which substituted for the canal wall down (CWD) procedure and involved slitting the posterior canal wall. The current, larger series compares the results of the CWW procedure with conventional surgical techniques. STUDY DESIGN: Retrospective analysis of 78 pediatric ears. SETTING: Academic tertiary referral center. PATIENTS: The mean patient age was 13.5 years. MAIN OUTCOME MEASURE(S): The data analyzed included ears later requiring conversion from CWW to CWD, dry/moist ear results, recidivation determined by two separate methods, and audiometric data statistically analyzed using independent samples analysis (unpaired, two-tailed Student's t test). RESULTS: First, of 42 CWW ear procedures, 6 (14%) later required conversion to CWD. Second, dry ear results were as follows: for CWW, 94%; for CWD, 92%; and for CWU (canal wall up), 90%. Third, recidivation determined at 1 year (standard rate) was, for CWW, 19.5%; for CWD, 0%; and for CWU, 7.7%; the at-risk calculation rate was, for CWW, 27%; for CWD, 0%; and for CWU, 8.3%. The 6-year recidivation rate for all three surgical techniques was 0%. 4). The mean preoperative-to-postoperative four-tone air-bone gap change was, for CWW, from 29.7 to 26.4 dB; for CWD, from 32.9 to 39.0 dB; and for CWU, from 21.0 to 25.2 dB (postoperative CWW to CWD, p < 0.005). A postoperative air-bone gap result of 0 to 20 dB was achieved as follows: with CWW, in 13 of 36 ears; with CWD, in 2 of 14 ears; and with CWU, in 9 of 22 ears. CONCLUSION: Frequently, a CWW procedure can be substituted for a traditional CWD procedure. In the extended series, the CWW technique continued to provide hearing results similar to CWU rather than to CWD procedures in a young population who will bear the surgical outcome for many decades. PMID- 15891651 TI - Long-term outcome of atticotomy for cholesteatoma in children. AB - OBJECTIVE: To evaluate the long-term outcome of atticotomy (AT) surgery for cholesteatoma in children. STUDY DESIGN: Case series, retrospective chart review. SETTING: General otology practice in a teaching hospital. PATIENTS: Fifty-three children aged 5 to 16 years old with cholesteatoma limited to the attic and middle ear who underwent AT between the years 1979 to 1994. INTERVENTIONS: AT consisting of removal of cholesteatoma confined mainly to the attic. The procedure consists of removal of the lateral attic wall (scutum) and involved ossicles with excision of the cholesteatoma and tympanic membrane grafting with temporalis fascia, leaving a small attical cavity. The mastoid antrum was left intact. MAIN OUTCOME MEASURES: Disease recurrence, middle ear status, hearing results, and the need for further surgery. RESULTS: Mean follow up was 5.1 (+/ 3.6) years. Eleven (21%) children required completion canal wall down tympanomastoidectomy because of residual disease or recurrence. The mean preoperative speech recognition threshold was 39.4 +/- 13.4 dB. The mean postoperative speech recognition threshold was 34.5 +/- 15.4, an improvement of 6.1 dB compared with the preoperative level. Last follow-up examination revealed a dry ear in all cases but one. CONCLUSIONS: In selected cases, AT can be a less invasive option in children with cholesteatoma limited to the attic and middle ear. PMID- 15891652 TI - Changes in blood neurotransmitter and steroid levels during evoked vertigo. AB - HYPOTHESIS AND BACKGROUND: Experimental evidence suggests that steroids as well as various neurotransmitters are critically involved in the functioning of the vestibular system in health and disease. Yet there are no pertinent human data. We hypothesized that changes in the serum levels of cortisol and plasma levels of excitatory and inhibitory neurotransmitters may occur during evoked vertigo. SUBJECTS AND METHODS: Ten healthy volunteers (median age 37, range 21-57) entered the study. Subjects were investigated at rest and at the time of maximal nystagmic reaction during caloric irrigation. The determination of glutamate, aspartate, and gamma-aminobutyric acid (GABA) was performed by reverse phase high performance liquid chromatography, whereas cortisol measurements were performed with an immunoenzymatic assay with fluorescence polarization. RESULTS: During evoked vertigo, cortisol levels increased from a baseline value of 11.86 (+/ 1.272) microg/dl to 14.375 (+/-2.183) microg/dl (p < 0.01), whereas all neurotransmitter levels decreased significantly. Glutamate levels, for instance, fell from a resting value of 25.99 (+/-6.30) ng/ml to 17.40 (+/-5.50) ng/ml (p < 0.001), and aspartate and GABA decreased as well. CONCLUSION: Evoked vertigo is consistently associated with an increase in steroid serum levels and accompanying decreases in the plasma levels of glutamate, aspartate, and GABA. The possible underlying mechanisms and the functional significance of these findings are discussed. PMID- 15891653 TI - Combined retrosigmoid retrolabyrinthine vestibular nerve section: results of our experience over 10 years. AB - OBJECTIVE: We aimed to evaluate the results of our experience in vestibular nerve sectioning (VN), which was performed using combined retrosigmoid retrolabyrinthine approach. STUDY DESIGN: Medical records of 280 patients who were consecutively operated on for incapacitating peripheral vertigo were retrospectively evaluated, and 210 patients who completed 2 years follow-up and had adequate follow-up data were found to be suitable for inclusion in the study. METHODS: Hearing results, vertigo control rates, and complications of the retrosigmoid-retrolabyrinthine VN were evaluated. RESULTS: The patients were suffering from vertigo for a mean period of 32.2 months. Bilateral Meniere's disease occurred in 5.7% of the patients in the follow-up period. A complete or substantial vertigo control could be achieved in 94.4% of the patients (191 [90.1%] in Class A and 9 [4.3%] in Class B). Preoperative speech reception threshold, pure-tone average, and speech discrimination score of the patients were 56.5 dB, 47.4 dB, and 73.6%, respectively. Postoperative corresponding values were 62.2 dB, 43.4 dB, and 68.5%, respectively (p > 0.05). The complication rate was low (2.5%). Most common complication was abdominal hematoma, which was seen in 4.5%. CONCLUSION: VN performed using retrosigmoid retrolabyrinthine approach has low complication and high vertigo control and hearing preservation rates. It can be applied as an initial surgery or reserved as the last step when the other surgical treatments have failed to control vertigo. PMID- 15891654 TI - Benign paroxysmal positional vertigo can interfere with the cardiac response to head-down tilt. AB - OBJECTIVE: To assess the pulse rate and the respiratory rate responses to head down tilt of the whole body in the plane of the posterior canals in healthy subjects and in patients with benign paroxysmal positional vertigo (BPPV). BACKGROUND: Although BPPV attacks are usually accompanied by autonomic symptoms, there are no studies assessing autonomic responses during triggering maneuvers for BPPV, neither in healthy subjects nor in patients. METHODS: We evaluated nine healthy subjects and four BPPV patients (3 unilateral and 1 bilateral). Using a two-axis rotator, from an upright position they were rotated 135 degrees backwards to head-down tilt, either in the plane of the right or the left posterior canal. RESULTS: In healthy subjects, head-down tilt always induced a significant decrease of the pulse rate, which was similar after rotation to the right and to the left posterior canals. This response was observed in patients with unilateral BPPV only when they were rotated toward the nonaffected side, and it was not evident when they were rotated toward the affected side (p < 0.025). In the patient with bilateral BPPV, no change of the pulse rate was observed after rotation toward the right or to the left posterior canal. Although, in all the patients, the respiratory rate increased during the tilt, a similar increase was observed in two healthy subjects. CONCLUSION: After rotation in the plane of the affected semicircular canal, BPPV can interfere with the cardiac response to head-down tilt of the whole body. PMID- 15891655 TI - Anatomic differences in the lateral vestibular nerve channels and their implications in vestibular neuritis. AB - HYPOTHESIS: Anatomic differences may render the superior division of the vestibular nerve more susceptible to injury during vestibular neuritis. BACKGROUND: Neural degeneration has been identified in temporal bone studies of vestibular neuritis. Previous anatomic and physiologic studies of vestibular neuritis have demonstrated that the superior division of the vestibular nerve is preferentially affected, with sparing of the inferior division. A preliminary temporal bone study has implicated neural entrapment as a possible cause for this preferential injury. METHODS: Two independent unbiased observers performed histologic analysis of 184 temporal bones from our temporal bone library. Measurements of the medial, midpoint, and lateral portions of the superior vestibular, inferior vestibular, and the singular nerves and their bony channels lateral to the internal auditory canal were made. These measurements included the length and width of each bony channel and an estimated percent of each channel occupied by bony spicules at each location. RESULTS: The lengths of the bony channels of the singular nerve (0.598 mm) and the inferior vestibular nerve (0.277 mm) were significantly shorter than the average length of the superior vestibular channel (1.944 mm; p < 0.0001). The total percent of the channel occupied by bone at the midpoint was significantly greater for the superior vestibular (28%) compared with either the singular (0%) or the inferior vestibular channel (18%) (p < 0.0001). CONCLUSION: The lateral bony channel of the superior vestibular nerve is seven times longer than the inferior vestibular and more than three times longer than the singular channel. There are a larger percentage of bony spicules occupying the superior vestibular compared with the inferior vestibular or singular channels. In addition, the superior nerve passes through a longer area of severe narrowing compared with the inferior or singular nerves. This anatomic arrangement of a longer bony channel with more interspersed bony spicules could make the superior vestibular nerve more susceptible to entrapment and ischemia. PMID- 15891656 TI - Quantitative study of the vestibular sensory epithelium in cochleosaccular dysplasia. AB - BACKGROUND: Cochleosaccular dysplasia is the most common pathologic finding seen in children with profound congenital sensorineural hearing loss. There has been no quantitative study on the peripheral vestibular system in cochleosaccular dysplasia. OBJECTIVE: To investigate quantitatively the extent of pathologic changes of the vestibular sensory epithelium in cochleosaccular dysplasia. SUBJECTS AND METHODS: Thirteen temporal bones with congenital deafness from 10 individuals were selected for this study from the temporal bone collection of University of Minnesota that showed suitable pathologic findings for the histopathologic criteria of cochleosaccular dysplasia. Age-matched normal control temporal bones were also selected. The vestibular hair cells including types I and II hair cells were counted separately in the saccular macula, utricular macula, and three cristae of the semicircular canals using Nomarski microscopy. RESULTS: The hair cell densities of types I and II hair cells in the macula of the saccule in cochleosaccular dysplasia were significantly decreased compared with the data of normal subjects. Both types I and II hair cells in the utricular macula and the cristae of the three semicircular canals in cochleosaccular dysplasia were well preserved, and no significant difference was observed between findings of cochleosaccular dysplasia and normal controls in the utricle and the three semicircular canals. CONCLUSIONS: In cases with cochleosaccular dysplasia, the neurosensorial hair cells of the saccule were affected; however, the osseous labyrinth, the membranous utricle, and the semicircular canals were normal. Further studies should be performed to establish the pathogenesis of cochleosaccular dysplasia in humans. PMID- 15891657 TI - Patient and normal three-dimensional eye-movement responses to maintained (DC) surface galvanic vestibular stimulation. AB - HYPOTHESIS: That disease or dysfunction of vestibular end organs in human patients will reduce or eliminate the contribution of the affected end organs to the total eye-movement response to DC surface galvanic vestibular stimulation (GVS). BACKGROUND: It was assumed that DC GVS (at current of 5 mA) stimulates all vestibular end organs, an assumption that is strongly supported by physiological evidence, including the activation of primary vestibular afferent neurons by galvanic stimulation. Previous studies also have described the oculomotor responses to vestibular activation. Stimulation of individual semicircular canals results in eye movements parallel to the plane of the stimulated canal, and stimulation of the utricular macula produces changes in ocular torsional position. It was also assumed that the total three-dimensional eye-movement response to GVS is the sum of the contributions of the oculomotor drive of all the vestibular end organs. If a particular vestibular end organ were to be diseased or dysfunctional, it was reasoned that its contribution to the GVS induced oculomotor response would be reduced or absent and that patients thus affected would have a systematic difference in their GVS-induced oculomotor response compared with the response of normal healthy individuals. METHODS: Three dimensional video eye-movement recording was carried out in complete darkness on normal healthy subjects and patients with various types of vestibular dysfunction, as diagnosed by independent vestibular clinical tests. The eye movement response to long-duration bilateral and unilateral surface GVS was measured. RESULTS: The pattern of horizontal, vertical, and torsional eye velocity and eye position during GVS of patients independently diagnosed with bilateral vestibular dysfunction, unilateral vestibular dysfunction, CHARGE syndrome (semicircular canal hypoplasia), semicircular canal occlusion, or inferior vestibular neuritis differed systematically from the responses of normal healthy subjects in ways that corresponded to the expectations from the conceptual approach of the study. CONCLUSION: The study reports the first data on the differences between the normal response to GVS and those of patients with a number of clinical vestibular conditions including unilateral vestibular loss, canal block, and vestibular neuritis. The GVS-induced eye-movement patterns of patients with vestibular dysfunction are consistent with the reduction or absence of oculomotor contribution from the end organs implicated in their particular disease condition. PMID- 15891658 TI - Acoustic schwannomas: awareness of radiologic error will reduce unnecessary treatment. AB - OBJECTIVE: To measure the intra- and interobserver error in size estimation of acoustic schwannomas from magnetic resonance imaging (MRI) scans by experienced radiologists to determine whether small amounts of tumor growth that may affect management (2 mm) could be reliably measured in clinical practice. DESIGN: Duplicated, blinded size estimation of acoustic neuromas (according to American Academy of Otolaryngology-Head and Neck Surgery guidelines, 1995) from MRI scans of patients with acoustic neuromas. SETTING: Tertiary referral teaching hospital and DGH. PARTICIPANTS: Four radiologists (including 2 dedicated neuroradiologists) measuring positive MRI scans of 26 patients with an acoustic neuroma. MAIN OUTCOME MEASURE: Intraradiologist and inter-radiologist repeatability coefficients in millimeters for the maximal tumor diameter in the anteroposterior (AP) axis, medial-longitudinal (ML) axis, and the square-root of the product of these two measurements. Repeatability coefficients give the 95% range within which the differences in repeated measurements lie. RESULTS: The intraradiologist repeatability for AP and ML measurements ranged from 1.51 to 6.03 mm and 2.01 to 3.83 mm, respectively. The repeatability of the square-root of the product ranged from 1.43 to 4.94 mm. The inter-radiologist repeatability was 6.48 mm and 7.46 mm for the AP and ML measurements, respectively, giving a repeatability of 3.65 mm for the square-root of the product. CONCLUSION: The study indicates that, in routine clinical practice, differences in tumor size of the order of 2 mm cannot be reliably measured, even by the same radiologist. Thus, reported growth of acoustic tumors should be interpreted with caution, especially if this is the criterion for recommending treatment. PMID- 15891659 TI - Facial paralysis and surgical rehabilitation: a quality of life analysis in a cohort of 1,595 patients after acoustic neuroma surgery. AB - OBJECTIVES: On the basis of survey results of the Acoustic Neuroma Association, we report patient ratings of facial dysfunction and outcomes for various facial rehabilitative therapies after surgical treatment of acoustic neuroma (AN). We assessed patients' perceived quality of life (QOL) and reviewed the literature regarding facial dysfunction and its management associated with AN. STUDY DESIGN: The Acoustic Neuroma Association mailed a detailed questionnaire to 2,372 members to identify preoperative and postoperative symptoms, complications, and long-term effects on physical and psychosocial function. A cohort of 1,595 (82.2%) respondents who underwent surgical treatment of ANs reported their experiences with facial dysfunction. PATIENTS: Of all 1,940 survey respondents, 1,682 of 1,875 that had ANs underwent surgical treatment. The study included 1,595 patients with ANs (82.2% of all respondents) who underwent surgical treatment by way of the translabyrinthine, suboccipital, or middle fossa approaches and excluded 87 respondents who did not report the type of surgical approach. METHODS: Respondents answered questions intended to qualify and quantify the degree that facial dysfunction impacted QOL parameters. Responses were analyzed for tumor size, surgical approach, patient age, and sex. Statistical analysis was performed using SPSS software. RESULTS: In our analysis, 11% of all respondents experienced some degree of preoperative facial weakness or eye problems. Of all respondents, 45.5% (725 patients) experienced worsened facial weakness caused by surgery, and of these, 72% reported that it was permanent. The most commonly used successful therapy for facial reanimation for 271 (19.6%) patients was placement of a gold weight. The factor most often associated with poor outcome was a large tumor. Of all respondents, 28% felt significantly affected by facial weakness, 63% felt their smile was symmetric, and 70% were content "quite a bit" or "very much" with their QOL. CONCLUSIONS: In this large cohort study of AN patients, facial dysfunction was a significant morbidity. Physicians should be aware of the risk factors identified, specifically large tumor size and the impact facial dysfunction has on QOL, when counseling patients regarding optimal management of AN. PMID- 15891660 TI - Closure of complex lateral skull base defects. AB - OBJECTIVE: To discuss surgical reconstructive options and complications in patients with extensive lateral skull base defects. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Eligibility criteria included patients seen between July 1999 and July 2003 with malignant neoplasms of the lateral skull base requiring surgical therapy, with resultant surgical defect not amenable to primary closure. INTERVENTION: Surgical extirpation of malignant skull base neoplasm requiring free tissue transfer or rotational flap closure. MAIN OUTCOME MEASURE: Closure and healing of surgical defect, intraoperative and postoperative complications, patient survival. RESULTS: There were 11 patients, 8 males and 3 females, with an average age of 65 years. Eight patients required trapezius flap reconstruction, whereas one patient required temporalis rotational flap closure, and two patients required rectus abdominus free tissue transfers. There was one perioperative death secondary to cardiac disease. There were no immediate wound complications. One patient developed a delayed partial trapezius flap failure successfully treated with a rectus abdominus flap. CONCLUSIONS: The trapezius rotational flap is a reliable means of closing complex lateral skull base defects with minimal morbidity. PMID- 15891661 TI - Vertebral artery aneurysm causing contralateral cerebellopontine angle mass effect. AB - OBJECTIVE: A vertebral artery aneurysm mimicking a contralateral cerebellopontine angle tumor is described. This is only the second reported case of this in the literature. STUDY DESIGN: Case report. SETTING: Radcliffe Infirmary, Oxford, United Kingdom. PATIENT: Seventy-year-old female patient who presented with a gradually deteriorating right-sided sensorineural hearing loss and a right facial palsy. INTERVENTIONS: Magnetic resonance imaging scan and computed tomography angiography with three-dimensional reconstruction followed-up by endovascular coiling of a vertebral artery aneurysm. MAIN OUTCOME MEASURE: Deterioration in neurologic status. RESULTS: Satisfactory recovery, and at 3 months, the patient is well with no deterioration in facial nerve function. CONCLUSION: Imaging by means of computed tomography angiography complements the standard magnetic resonance imaging scan in the rare event of an aneurysm at the cerebellopontine angle. Endovascular coiling occlusion is a recognized and effective way of managing intracranial aneurysms at the cerebellopontine angle. PMID- 15891662 TI - Delivery of neurotrophin-3 to the cochlea using alginate beads. AB - OBJECTIVE: The aim of this study was to design a novel cochlear neurotrophin (NT) delivery system for the rescue of auditory neurons after ototoxicity-induced deafening. BACKGROUND: NT-3 is a trophic growth factor that promotes the survival of the auditory nerve and may have a potential therapeutic role in slowing neuron loss in progressive deafness, especially as an adjunct to the current cochlear implant. Beads made from alginate are biodegradable, slow release substances that can be placed at the round window or inside the cochlea. This study investigated the loading properties, release kinetics, and implantation potential of alginate beads loaded with NT-3. METHODS: Alginate beads were prepared using an ionic gelation technique and postloaded with NT-3. Release of NT-3 was measured using enzyme-linked immunosorbent assay over 5 days. Alginate beads were implanted into deafened guinea pigs for 28 days, after which survival of auditory neurons was assessed. RESULTS: Enzyme-linked immunosorbent assay studies demonstrated a 98% to 99% loading of NT-3 with a slow, partial release over 5 days in Ringer's solution. Furthermore, the addition of heparin to the delivery system modulated NT-3 release to a steadier pattern. Implantation of alginate-heparin beads in guinea pig cochleae produced minimal local tissue reaction. NT-3 loaded beads implanted at both the round window and within the scala tympani of the basal turn provided auditory neurons significant protection from degradation and apoptosis compared with unloaded beads or untreated cochleae. CONCLUSIONS: This study demonstrates alginate beads to be a safe, biodegradable and effective delivery system for NT-3 to the cochlea. PMID- 15891663 TI - Modification of atrial natriuretic peptide receptor expression in the rat inner ear. AB - HYPOTHESIS: The purpose of this animal study was to confirm the presence of all three atrial natriuretic peptide (ANP) receptor subtypes in the rat inner ear and compare the expression of each receptor after inner ear injection of ANP, phosphate-buffered saline, or a solution containing ANP incubated with anti-ANP antibody (to block upregulation). BACKGROUND: Receptors for ANP and related compounds have been localized in the inner ear of animals and humans. A previous study at this institution demonstrated the ability to up-regulate the expression of the three ANP receptors (ANP-A, ANP-B, ANP-C) in response to round window injection of ANP in the rat inner ear. METHODS: After surgical exposure, the round window of female Lewis rats was injected with various concentrations of ANP, ANP plus anti-ANP antibody, or control. Animals were killed 24 hours after injection, inner ear tissues were harvested and homogenized, and RNA was isolated for reverse-transcription polymerase chain reaction. RESULTS: Electrophoresis showed the presence of all three receptor subtypes with exposure to phosphate buffered saline. Expression was significantly higher 24 hours after injection with the two concentrations of ANP. This increase was partially blocked with increasing relative concentrations of anti-ANP antibody. CONCLUSIONS: These findings confirm the presence and responsiveness of ANP receptors in the rat inner ear. The ability to block up-regulation with the antibody provides a potential new research tool for manipulating the function of this hormone system in experimental models and, ultimately, in understanding the mechanisms of fluid homeostasis in the inner ear. PMID- 15891664 TI - Sonotubometry: eustachian tube ventilatory function test: a state-of-the-art review. AB - OBJECTIVE: Disturbance of any of the ET functions may contribute to the development of otitis media. Sonotubometry measures the ventilatory function using sound. The qualities of sonotubometry as a test for eustachian tube ventilatory function have been studied by various investigators. The development of the method is described in the review, and a summary of the study results is provided to make an estimate of the diagnostic potential of this eustachian tube function test. DATA SOURCES: The English-language literature on the topic was searched systematically by Medline and Pubmed using the following key words: ventilatory function, eustachian tube, sonotubometry, and function test. There were no limits for the year of publication. STUDY SELECTION: Articles that described the method itself (validity, reproducibility, diagnostic value) were studied in detail. DATA EXTRACTION: All the articles described in study selection were used for this review. CONCLUSIONS: The technique of sonotubometry has been improved gradually over the years. The results of sonotubometry are at least as good as those of other function tests. However, because the results still tend to be ambiguous in children and otitis media is most common in this population, the reproducibility and application of sonotubometry must be evaluated further. Sonotubometry has great advantages over other function tests, but it is not used routinely to assess eustachian tube ventilatory function because its value for clinical practice has not yet been adequately demonstrated. The review showed that sonotubometry can be improved further and that efforts to do so seem justified because it forms a particularly promising method to assess eustachian tube function in children with suspected eustachian tube disease. PMID- 15891665 TI - Michel's aplasia. PMID- 15891666 TI - Wegener's granuloma. PMID- 15891667 TI - Radiologic follow-up after vestibular schwannoma surgery. PMID- 15891668 TI - Thomasz Drobnik: great Polish surgeon and patriot. PMID- 15891669 TI - Intracerebral abscess after BAHA abutment change. PMID- 15891671 TI - Diagnostic test for Meniere's disease. PMID- 15891673 TI - History of tympanoplasty. PMID- 15891674 TI - Meta-analysis of intratympanic gentamicin. PMID- 15891675 TI - Mayo Clinic office visit. Learning more about osteopenia. PMID- 15891676 TI - Restless legs syndrome. Treatments can ease troubling symptoms. PMID- 15891677 TI - High doses of vitamin E may increase risk of death. PMID- 15891678 TI - Olive oil may reduce coronary artery disease risk. PMID- 15891679 TI - Early symptoms of ovarian cancer exist but may not be recognized. PMID- 15891682 TI - Preparing for surgery. What to expect, how to plan. PMID- 15891680 TI - The inflammation and disease connection. PMID- 15891683 TI - Stress. Can it be positive? PMID- 15891684 TI - Can you tell me about the new device the FDA has approved for treating uterine fibroids? PMID- 15891685 TI - I've heard that drinking noni juice may lower my cholesterol. Could this be true? PMID- 15891687 TI - Preventing cancer. A healthy lifestyle can significantly reduce your risk. PMID- 15891688 TI - Molecular breast imaging detects small tumors. PMID- 15891689 TI - Alternative to tamoxifen effective for preventing breast cancer recurrence. PMID- 15891690 TI - Women's Health Week offers reminder to get checkups, preventive screening. PMID- 15891691 TI - Recovering from a stroke. A team approach can improve outcomes. PMID- 15891693 TI - Being underweight. A look at causes, when to seek help. PMID- 15891694 TI - Yogurt. How good it is. PMID- 15891695 TI - I want to use a mail-order pharmacy, but I've heard medications can be damaged when they're sent through the mail. Is this true? PMID- 15891696 TI - Should women in their 80s and 90s still get screening mammograms? PMID- 15891697 TI - How can I keep from getting an insect-borne illness? PMID- 15891698 TI - Opportunities to improve outcomes in the heart failure patient: alleviating symptoms and prolonging life. Introduction. PMID- 15891699 TI - Implications of heart failure drug trials: COMET, CHARM, EPHESUS. AB - Activation of the renin-angiotensin-aldosterone and adrenergic nervous systems plays a major role in the progression of heart failure, and inhibitors and antagonists of these neurohormonal systems improve outcomes. b-Blockers, angiotensin-converting enzyme inhibitors, and aldosterone antagonists have been shown to improve parameters such as ventricular remodeling, ejection fraction, and renal function and to reduce rates of morbidity and mortality. This article reviews 3 recent clinical trials that have added to our knowledge of the use of these agents. Two of the studies-EPHESUS and COMET-demonstrated significant reduction in all-cause mortality, whereas the third-CHARM-showed a marginal reduction. These trials established that it is feasible to design and execute heart failure studies of sufficient scale to assess improvement in rates of mortality and morbidity. PMID- 15891700 TI - Sudden cardiac death: epidemiology and temporal trends. AB - Sudden cardiac death (SCD) now accounts for more than half of all coronary heart disease deaths in the United States. The majority of cases are due to underlying coronary artery disease, and deaths from both coronary artery disease and SCD have declined markedly over the past several decades due to improved primary and secondary prevention and treatment strategies. This review examines the current statistics on the prevalence of SCD, and identifies those patients at greatest risk. It also discusses existing tests and treatments, including medication that results in neurohormonal antagonism, and devices such as the implantable cardioverter defibrillator (ICD) and cardiac resynchronization therapy with a defibrillator (CRT-D). Along with increased public awareness of SCD as a major health risk, physicians are advised to implement proven effective drug and devices that can improve survival. PMID- 15891701 TI - Device trials in heart failure: a focused summary. AB - Despite considerable progress in heart failure management with pharmacologic agents, measures to bring about significant improvements in morbidity and mortality are still needed. Cardiac resynchronization therapy (CRT) is a means to enhance myocardial function by stimulating the failing left ventricle at or near the time of right ventricular activation to synchronize ventricular depolarization. Current data from randomized, controlled trials suggest that CRT benefits patients with moderate to severe heart failure and have shown that this therapy significantly reduces mortality and hospital admissions in this group. In addition to CRT, implantable cardioverter-defibrillators have been evaluated in heart failure patients with significantly reduced left ventricular function and have been shown to reduce mortality from sudden cardiac death. This article summarizes recent device trials and discusses how best to apply their results to clinical practice. PMID- 15891702 TI - Comprehensive treatment of heart failure: state-of-the-art medical therapy. AB - Despite advances in therapy and better outcomes for heart failure, this disease remains burdensome in terms of hospitalization costs, quality of life, and mortality. Many treatment strategies are available for heart failure, including medical therapy with agents such as angiotensin-converting enzyme inhibitors, and b-blockers, and device therapy including implantable cardioverter-defibrillators and cardiac resynchronization. However, data now demonstrate that compliance with these evidence-based strategies is well below acceptable thresholds, negatively affecting quality of care. The implementation of guidelines such as those of the American College of Cardiology/American Heart Association and the application of dedicated disease management programs are two mechanisms aimed toward helping physicians construct and adhere to effective treatment regimens for their patients with heart failure. PMID- 15891703 TI - Strategies to improve the use of evidence-based heart failure therapies. AB - Patients with heart failure (HF) face a very high risk of hospitalizations, arrhythmias, and mortality. Despite compelling scientific evidence that angiotensin-converting enzyme inhibitors, b-blockers, and aldosterone antagonists reduce hospitalizations and mortality in HF patients, these life-prolonging therapies continue to be underutilized. Recently, device therapy for HF, including implantable cardioverter-defibrillators and cardiac resynchronization devices, has also been demonstrated to result in substantial mortality reduction. A number of studies in a variety of clinical settings have documented that a significant proportion of HF patients are not receiving treatment with these guideline-recommended, evidence-based therapies when guided by conventional care. Treatment gaps in providing other components of HF patient care, including patient education, have also been documented. The demonstration that initiation of cardiovascular protective medications before hospital discharge results in a marked increase in treatment rates, improved long-term patient compliance, and better clinical outcomes has led to the revision of national guidelines to endorse this approach as the standard of care. Recent studies demonstrated that hospital-based systems can improve medical care and education of hospitalized HF patients and accelerate use of evidence-based, guideline-recommended therapies by administering them before hospital discharge. HF disease management programs have also been shown to improve HF treatment, resulting in substantial reduction in hospitalizations and mortality. Further efforts are needed to ensure the implementation of effective strategies and systems that increase the use of evidence-based therapies in the hospital and outpatient settings to reduce the substantial HF morbidity and mortality risk. PMID- 15891706 TI - National Institute of Nursing Research working group on "Optimizing pregnancy outcomes in minority populations". AB - The growing complexity of biomedical research requires new methods of discovery; scientists must use an interdisciplinary approach and explore new models of team science, as underscored in the Roadmap of the National Institutes of Health. In March 2003, the National Institute of Nursing Research convened a working group of scientists and clinicians with a wide range of backgrounds to address "Optimizing Pregnancy Outcomes in Minority Populations." The 2-day meeting included a variety of presentations on the current state of research on pregnancy in minority populations. Many participants provided specific insights regarding biobehavioral issues in human-environment interaction, stress and health status relationships to risk, maternal-fetal interactions, and the complications of pregnancy. This supplement presents articles from several participants at this interdisciplinary meeting. The National Institute of Nursing Research looks forward to further collaborations across the National Institutes of Health and other agencies to achieve the vital aims of this working group. PMID- 15891707 TI - Environmental exposures, toxicologic mechanisms, and adverse pregnancy outcomes. AB - Environmental risk factors (defined as those agents and stresses that are generally the responsibility of environmental agencies) are often tangible indicators of economic and social disparity in the United States. Many site specific analyses have reported that communities of color and poverty are exposed more often and more intensively to such environmental hazards as lead, air pollution, agrochemicals, incinerator emissions, and releases from hazardous waste sites. Thus, exposures to these toxicants may explain part of the socioeconomic disparity that is observed in terms of risks of adverse pregnancy outcomes. The purpose of this study was to describe the associations between certain environmental exposures and reproductive outcomes through a discussion of both epidemiologic and animal model studies. In addition, we list potential sources of exposure data and describe physiologic changes in pregnancy that may increase the likelihood of both external exposures and increased internal dose. Several models for further study of environmental risk factors are suggested to increase our understanding of gene-environment interactions toward the goal of indentifying preventable risk factors to improve reproductive outcomes of particular concern to disadvantaged populations. PMID- 15891708 TI - Neighborhood context and reproductive health. AB - Racial and social class differences in rates of preterm birth and other adverse outcomes are among the most widely recognized and least well-understood phenomena in the study of reproductive health. Individual-level characteristics have failed to account for such gradients. Recently, researchers have begun to argue that health in general and reproductive outcomes specifically are rooted in social inequalities. One area of such inequality may be residential segregation and the associated race/ethnic differences in exposure to adverse neighborhood conditions. We review the empiric data that examine the association between neighborhood conditions and reproductive health. We also review the major challenges that researchers face when trying to incorporate neighborhood-level variables into studies of health outcomes. Our goal is to stimulate further research that simultaneously considers social, economic, and biologic determinants of reproductive health. PMID- 15891709 TI - Overview of current state of research on pregnancy outcomes in minority populations. AB - Pregnancy outcomes improved significantly over the 20th century in the United States but currently vary widely between women of different ethnic and racial backgrounds. The current health disparities that exist are based, in part, only on differences in socioeconomic status or education. There is wide variability in pregnancy outcomes within specific subgroups of women. Disparities may be due to underlying differences in health before pregnancy, differences in community norms, and individual lifestyle choices and to differences in health care delivery systems. Areas for needed research and promising new models of care are reviewed. PMID- 15891710 TI - Psychosocial stress and neuroendocrine mechanisms in preterm delivery. AB - This review focuses on the contribution of psychosocial stress to the racial/ethnic disparities in preterm delivery in the United States and addresses the subset of psychosocial stressors that are disproportionately prevalent among minority women. We argue that chronic exposure to poverty, racism, and insecure neighborhoods may condition stress responses and physiologic changes in ways that increase the risk of preterm delivery. Cumulative stressors may impact pregnancy outcomes through several intersecting pathways, which include neuroendocrine, behavioral, immune, and vascular mechanisms. Many of these pathways also lead to chronic disease. It may be useful to consider preterm delivery as a chronic disease with roots in childhood, adolescence, and early adulthood. Like other physiologic systems, the female reproductive axis may be vulnerable to the physiologic "wear and tear" of cumulative stress, which results in preterm delivery. PMID- 15891711 TI - Biochemical markers for the prediction of preterm birth. AB - The prediction of preterm birth may be important (1) to initiate risk specific treatment; (2) to define a population that is at risk in which to study a particular treatment; or (3) to better understand the pathways that lead to preterm birth. Biologic fluids that have been used as sources for tests include serum, plasma, amniotic fluid, urine, vaginal and cervical secretions, saliva, and even periodontal fluid. We discuss the types of substances that are found in body fluids (eg, organisms, cytokines, enzymes, hormones) that have been studied as predictors of preterm birth, the fluids in which they are found, and issues that are related to the timing of the test, the cost, and the ease of fluid collection and processing. We emphasize that a test for any of these substances should not be introduced into clinical practice until the use of the test, which is followed by an appropriate intervention, leads to a reduction in preterm birth. PMID- 15891712 TI - Stress model for research into preterm delivery among black women. AB - The disparity between black and white infant mortality rates increased over the last decade, despite overall improvement in infant survival. Because most black infant deaths are related to preterm delivery, the discovery of the cause of premature birth in general and excess premature birth for black infants in particular is of paramount importance for reproductive health research. Substantial theoretic support exists for maternal stress as a risk factor for preterm birth. Traumatic events early in life may sensitize the adult to contemporary stresses and increase her vulnerability to stress-induced neuroendocrine or infection/inflammatory pathways to early parturition. In addition, an individual may prematurely age as a result of cumulative stress or a major traumatic event. This "stress age," which is synonymous with the concept of weathering and similar to the concept of allostatic load, may affect parturition through chronic conditions (such as hypertension) and in poorly understood pathophysiologic mechanisms that are related to increased chronologic age. One potential measure of stress age is maternal serum dehydroepiandrosterone sulfate. Maternal stress is a potential explanatory factor for excess preterm delivery among black women because of their exposure to racism-associated stress. However, few studies have addressed this question, and results are mixed. Future etiologic research must take into account the complexities of the measurement of stress age and past and current exposures to stress, which includes internalized racism and interpersonal racism. PMID- 15891713 TI - Racial inequalities in preterm delivery: issues in the measurement of psychosocial constructs. AB - Psychosocial risk factors for preterm delivery have been a topic of much recent research. By extension, the role of psychosocial risks in the explanation of the racial and ethnic gap in preterm birth is of increasing interest. Several barriers that are related to the measurement of psychosocial variables may preclude a sound understanding of their role in the explanation of the racial gap in preterm birth. The purpose of this study was to present selected principles of measurement that should be addressed during the investigation of the relationship between psychosocial factors and preterm delivery to gain a greater understanding of the racial gap in preterm birth. PMID- 15891714 TI - Research strategies for optimizing pregnancy outcomes in minority populations. AB - The elimination of disparities in pregnancy outcomes is a common goal of clinicians and scientists and requires the collaboration of many disciplines to address the complexities of this still-increasing perinatal health concern. This commentary synthesizes the presentations and dialogue from a multidisciplinary workgroup meeting that was sponsored by the National Institute of Nursing Research in 2003. Concepts that are central to our understanding of the development and expression of such disparities are summarized, and approaches that are recognized as important in multiple disciplines that include basic, clinical, and social sciences are presented. Research strategies to foster a multidisciplinary research agenda are presented as a basis for future endeavors to improve pregnancy outcomes. PMID- 15891715 TI - Tissue-engineered collagen meniscus implants: 5- to 6-year feasibility study results. AB - PURPOSE: In this feasibility study, a 5- to 6-year clinical follow-up evaluation was conducted on 8 patients who had undergone reconstruction of 1 injured medial meniscus with a tissue-engineered collagen meniscus implant. The hypothesis was that these patients would show significant clinical improvement over their preoperative status and would have maintained their status determined at the 2 year follow-up evaluation. TYPE OF STUDY: Prospective longitudinal feasibility study follow-up evaluation. METHODS: Eight patients underwent arthroscopic placement of a collagen meniscus implant by a single surgeon to reconstruct and restore the irreparably damaged medial meniscus of 1 knee. All patients returned for clinical, radiographic, magnetic resonance imaging, and arthroscopic examinations an average of 5.8 years (range, 5.5-6.3 y) after collagen meniscus implant placement. RESULTS: Lysholm scores improved significantly (P = .045) from 75 preoperatively to 88 at most recent follow-up evaluation. Average Tegner activity scores improved significantly (P = .001) from 3 to 6. Patient self assessment improved significantly (P = .046) from 2.4 to 1.9 (1 = normal, 4 = severely abnormal). Pain scores improved from 23 to 11 (0 = no pain, 100 = worst pain). Imaging studies confirmed that the chondral surfaces of the medial compartment had not degenerated further since the placement of the implant 5.8 years earlier. Relook arthroscopy with direct measurement of the newly generated tissue revealed 69% defect filling. Histologic assessment of tissue biopsy specimens from 3 patients showed the presence of fibrocartilage with a uniform extracellular matrix. CONCLUSIONS: The meniscus-like tissue that developed after collagen meniscus implant placement has maintained its structure and functioned without negative effects for more than 5 years. The hypothesis was affirmed that these patients were improved significantly compared with their preoperative status and unchanged compared with 2-year evaluations. LEVEL OF EVIDENCE: Level IV. PMID- 15891716 TI - Long-term results of lateral retinacular release. AB - PURPOSE: We evaluated the outcomes of lateral retinacular release (LRR) after a long-term follow-up period of 5 to 12 years. TYPE OF STUDY: Long-term retrospective clinical follow-up study. PATIENTS AND METHODS: Between 1986 and 1994, 120 LRRs were performed in the Orthopaedic Department of the Catholic University of Rome. A total of 100 patients were evaluated. We divided the patients into 2 groups: group I contained 50 patients with patellar pain and no signs of instability; the remaining 50 patients, with clear signs of patellar instability, made up group II. Standard weight-bearing radiographs, axial views of the knee at 45 degrees , and dynamic computed tomography scans were performed in all patients preoperatively and at follow-up evaluation. Chondral damage was classified at the time of lateral release according to the criteria of Outerbridge and Dunlop. We used the Lysholm II score, which was modified for patellofemoral pathology and a clinical grading system of Busch and de Haven, to evaluate clinical outcomes at follow-up evaluation. RESULTS: In group I (pain), 70% reported satisfactory outcomes at follow-up evaluation compared with 50% in group II (P < .05) (instability). Compared with a previously published analysis of 3-year outcomes in this same patient population, there was very little change in group I patients, whereas group II showed a significant decrease in good outcomes over time. The worst results were obtained in cases with serious cartilage damage and exposure of the subchondral bone at the time of lateral release. CONCLUSIONS: LRR is a procedure offering a good percentage of success in the management of a stable patella with excessive lateral pressure and elective location of pain on the lateral retinaculum. In patellar instability the results are less favorable in long-term follow-up evaluation. The presence of high-grade joint surface injury is a poor prognostic indicator for lateral release. LEVEL OF EVIDENCE: Level IV. PMID- 15891717 TI - Evaluation of bone plug and soft tissue anterior cruciate ligament graft fixation over time using transverse femoral fixation in a sheep model. AB - PURPOSE: To evaluate the mechanical role of bone-anterior cruciate ligament (ACL) graft junction in comparison with primary fixation of the graft. TYPE OF STUDY: Ex vivo controlled biomechanical study. METHODS: An ACL reconstruction was performed on 2 groups of 40 sheep each. The ACL graft was patellar tendon in group 1, and free tendon in group 2. Load-to-failure tests were performed at 1, 2, 3, and 6 months. Each group was divided into 2 subgroups: In subgroup A the femoral fixation devices were removed before testing and in subgroup B they were left in place. RESULTS: At 1 month, structural properties of subgroup 2A were significantly lower than other subgroups. Until the third month, structural properties of each subgroup were significantly lower than normal ACL. Grafts always failed at midsubstance, except for subgroup 2A at 1 month, which pulled out of the femoral tunnel. CONCLUSIONS: Bone plug incorporation was stronger than graft strength at 1 month whereas soft tissue tendon incorporation was not stronger than graft until 2 months. At 3 months, both groups failed at approximately 30% of the native ACL strength, and at 6 months, both groups were nearly equal in terms of load to failure and stiffness. CLINICAL RELEVANCE: High fixation strength and stiffness of ACL graft does not allow earlier return to sports activities. PMID- 15891718 TI - Differential pretensions of a flexor tendon graft for anterior cruciate ligament reconstruction: a biomechanical comparison in a porcine knee model. AB - PURPOSE: The best way to apply the pretension force to the soft tissue graft during anterior cruciate ligament (ACL) reconstruction remains controversial. We compared manual, intra-, and extra-articular pretension techniques and determined the magnitude of displacement of the femur-graft-tibia complex in response to repetitive loading. TYPE OF STUDY: A biomechanical-controlled study. METHODS: Fresh porcine knees and profundus digital flexor tendons of hind limbs were used. Specimens were divided into 3 groups. Group A (manual pretension, n = 9) consisted of grafts that were pretensioned only by a maximal manual pull before final fixation on the tibial side. Group B (extra-articular pretension, n = 9) consisted of grafts that were pretensioned using a commercial tensiometer at 89 N for 15 minutes on the tendon preparation board, followed by pulling using maximal manual force before final fixation. Group C (intra-articular pretension, n = 9) consisted of grafts that were fastened on the femoral side first and then underwent application of a pretension force at 89 N for 5 minutes before final fixation on the tibial side. These femur-graft-tibia complexes from the 3 groups were loaded from 0 to 150 N at a frequency of 1 Hz for 1,000 cycles and then underwent a tensile load-to-failure test at a rate of 150 mm/min. RESULTS: The displacement of the femur-graft-tibial complex in response to cyclic loading for group C (5.4 +/- 0.3 mm) was significantly lower than those for groups A (12.5 +/ 1.1 mm) and B (8.8 +/- 0.8 mm) (P < .001). The fixation stiffness of group C (47.9 +/- 17.6 N/mm) was significantly greater than that of group A (32.5 +/- 9.7 N/mm) (P < .05) but not significantly different from that of group B (53.1 +/- 9.1 N/mm). The ultimate failure load in each group was not significantly different. CONCLUSION: Intra-articular pretension of the graft before final fixation can significantly minimize graft elongation at time 0 compared with manual and extra-articular pretension. CLINICAL RELEVANCE: Intra-articular or in vivo pretension of the graft using instruments may minimize the graft elongation in the early period of rehabilitation after ACL reconstruction. PMID- 15891719 TI - The sartorial branch of the saphenous nerve: its anatomy at the joint line of the knee. AB - PURPOSE: Injury to the saphenous nerve accounts for the majority of neurovascular complications during arthroscopic meniscal repairs. Although the anatomy of the infrapatellar branch has been studied, the location of the sartorial branch at the level of the joint line has not been defined adequately. The purpose of this study was to define the location of the sartorial branch in relation to surrounding structures at the level of the knee joint/meniscus. TYPE OF STUDY: Cadaveric anatomic study and imaging study. METHODS: We dissected 42 cadaveric knees and studied an additional 100 knees with axial magnetic resonance imaging (MRI). We measured distances from the nerve to adjacent structures and evaluated and graphically recorded 2-dimensional axial relationships. Dissections and MRIs were performed with the knee in full extension. RESULTS: In the dissected cadaveric knees, the distance from the nerve to the anterior border of the sartorius averaged 16.0 mm, with 90% greater than 10.0 mm. The nerve was extrafascial at the joint line in only 43% of the specimens. Axial MRI examinations found the average distance to the anterior border of the sartorius to be 16.5 mm, with 82% greater than 1.0 cm. The average distance to the gracilis tendon was 9.4 mm; 91% were within 1.0 cm and 51% within 5 mm. The average distance to the semitendinosus was 14.9 mm; none was within 5 mm. The nerve was never posterior or lateral to the semitendinosus, never posterolateral or lateral to the gracilis, and was anterior to the sartorius in only 3% of knees. CONCLUSIONS: This study describes the most common location of the sartorial branch of the saphenous nerve and the potential variability that may be encountered. CLINICAL RELEVANCE: Understanding the anatomy of the sartorial branch at the level of the joint line and its relationships to surrounding structures will help to minimize the risk of injury to the nerve during common surgical procedures at the posteromedial corner of the knee. PMID- 15891720 TI - Effect of arthroscopy and continuous cryotherapy on the intra-articular temperature of the knee. AB - PURPOSE: The purpose of this study was to determine "in vivo" the intra-articular temperature of the knee in baseline situation, to quantify its variation during the arthroscopic surgery and the repercussion on the body temperature, and to quantify the variation of the intra-articular temperature after the application of topic cold postoperatively. TYPE OF STUDY: Case series. Experimental study in patients who undergo arthroscopic surgery of the knee monitoring their intra articular temperature. METHODS: An intra-articular temperature data acquisition system is used in 30 patients who undergo arthroscopic surgery of the knee using saline solution at room temperature and monitoring their body temperature with a rectal temperature probe. In 23 of these patients, after surgery, a plastic bag with 2 kg of ice is applied on the knee, and the variation of the intra-articular temperature is measured for at least 1 hour. RESULTS: The basal temperature of the knee (32.22 degrees +/- 0.28 degrees C) decreases during the intervention using saline fluid at room temperature to nearly equalize it (24.49 degrees +/- 0.97 degrees C), and it led to a drop of the body temperature of 0.14 degrees +/- 0.04 degrees C. The continuous external application of ice after the arthroscopy produces a drop of the intra-articular temperature directly related to the time, with a magnitude of almost 4 degrees C in the first hour. CONCLUSIONS: The intra articular baseline temperature of the knee is significantly lower than that of the body, and the repercussion of its drop during arthroscopic surgery on the body temperature is insignificant. The external application of ice is an effective and sure method to diminish the intra-articular temperature of the knee. LEVEL OF EVIDENCE: Level IV, case series with no control group. PMID- 15891721 TI - Magnetic resonance imaging of the knee in asymptomatic professional basketball players. AB - PURPOSE: The purpose of this study was to evaluate the prevalence of articular cartilage lesions and meniscal tears on magnetic resonance imaging (MRI) scans in the knees of asymptomatic male professional basketball players. TYPE OF STUDY: A retrospective review. METHODS: Twenty players (40 knees) met the inclusion criteria of being signed by a National Basketball Association team and passing their preseason physical examination. All included athletes were assessed for the presence of articular cartilage lesions, meniscal pathology, and the presence of effusions. RESULTS: The overall prevalence of articular cartilage lesions on MRI was 47.5% in our study group. There were trochlear groove articular lesions in 25%. The lateral femoral condyle was involved in 2.5% of all knees. The medial femoral condyle was affected in 10% of all knees. The lateral tibial plateau showed articular cartilage lesions in 5%. The patella had articular cartilage lesions in 35%. The overall prevalence of various grade meniscal lesions was 20% on MRI. Medial intra-meniscal signals accounted for 87.5% and 12.5% on the lateral side. CONCLUSIONS: The results of our study show an equal to or higher prevalence of meniscal lesions in male professional basketball players than previously reported in the literature. We found a large number of patella-femoral articular cartilage lesions in our study population of male professional basketball players. These athletes perform at the highest demand level, which indicates that the presence of these lesions did not cause any symptoms. LEVEL OF EVIDENCE: Level IV, case series. PMID- 15891722 TI - Shared decision making, preoperative expectations, and postoperative reality: differences in physician and patient predictions and ratings of knee surgery outcomes. AB - PURPOSE: The present study was performed to determine the extent to which physicians and patients rate preoperative and postoperative knee pain and function differently, and to determine whether physicians or patients more accurately predict postoperative knee pain and function. TYPE OF STUDY: Longitudinal, prospective study. METHODS: Ninety-eight patients requiring either anterior cruciate ligament reconstruction surgery or meniscectomy and related surgery were interviewed 1 week before surgery, as well as 3 and 24 weeks postoperatively. Patients and their physicians completed ratings on knee pain and function at each time point. In addition, at their preoperative visit, patients and physicians completed ratings predicting their postoperative pain and functional status. RESULTS: Physicians rated patients as having less pain and greater knee function preoperatively and at 24 weeks postoperatively. Patients had more significant differences between predicted and actual ratings. CONCLUSIONS: Physicians tended to underestimate knee pain and overestimate knee function compared with patients. However, physicians better predicted postoperative knee pain and function ratings than did patients. These findings suggest that physician-patient discussions about preoperative expectations and postoperative reality might be an important part of clinical care. LEVEL OF EVIDENCE: Level II, Prospective Longitudinal Study. PMID- 15891723 TI - Split-line orientation of the talar dome articular cartilage. AB - PURPOSE: The purpose of this study was to define the pattern of superficial collagen fibril orientation on the talar dome and to create a template for rotational graft alignment during mosaicplasty. TYPE OF STUDY: An anatomic, cadaveric study. METHODS: Eight fresh-frozen human cadaveric tali had the articular surface of the talar dome pierced in grid fashion with a dissecting needle dipped in India ink. The longitudinal split that occurred between the superficial collagen fibers defined the fiber arrangement at each site of the talar dome. RESULTS: There was a remarkably consistent pattern to the split lines from specimen to specimen. CONCLUSION: Proper orientation of mosaicplasty plugs during transfer from the donor to recipient site with respect to superficial collagen fiber arrangement may allow the graft to better resist tensile stresses and may contribute to increased long-term graft longevity. CLINICAL RELEVANCE: Use of the talar dome map, combined with knowledge of the split-line orientation of the donor site, should allow surgeons performing mosaicplasty procedures on the talus to properly rotate their plugs with respect to split-line orientation. PMID- 15891724 TI - Arthroscopic-assisted surgical treatment for developmental dislocation of the hip. AB - PURPOSE: Treatment of developmental dislocation of the hip (DDH) includes surgical management in older children or in those who cannot be treated conservatively. However, complication rates of surgical treatment are quite high. The purpose of this report is to introduce our new surgical technique that can eliminate existing pathologic changes in DDH. TYPE OF STUDY: A small case series. METHODS: We performed arthroscopic-assisted surgical treatment in 4 hips of 4 female children who had DDH and no previous treatment. Their ages ranged from 11 to 14 months. Closed reduction under general anesthesia was tried just before the surgical intervention, but it failed in all of them. Tightness of the iliopsoas tendon was released followed by dissection of capsular adhesions using an anterolateral mini-incision. Excision of the hypertrophic ligamentum teres, transverse acetabular ligament, and pulvinar tissue was carried out using a double-portal arthroscopic procedure. We performed percutaneus adductor tenotomies in 2 cases. A spica cast and abduction splint were used for 12 to 17 weeks postoperatively. The follow-up of the patients was a minimum of 1 year. Although a 1-year follow-up period is adequate to evaluate the short-term results, it has been considered that there is a need for further studies that include long-term follow-up. We used the acetabular index and Shenton's line for preoperative and postoperative radiologic evaluation. Also, the cases were evaluated postoperatively in respect to range of motion restriction and the leg length discrepancy. RESULTS: The mean follow-up was 13.7 months (range, 12 to 16 months). Acetabular index measurements of the cases in the preoperative/postoperative periods were as follows: in the first case, 34 degrees/27 degrees; in the second case, 35 degrees/22 degrees; in the third case, 52 degrees/39 degrees; and in the fourth case, 40 degrees/28 degrees. Hip joint restriction and leg length discrepancy were not observed postoperatively. CONCLUSIONS: All the intra-articular structures (hypertrophic ligamentum teres, transverse acetabular ligament, and pulvinar tissue) in the acetabulum that impede the reduction of the femoral head have been eliminated by using the arthroscopic technique. The arthroscopic-assisted surgical treatment of DDH is successful in the short-term follow-up period. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 15891725 TI - Acetabular labral tears with underlying chondromalacia: a possible association with high-level running. AB - PURPOSE: The use of hip arthroscopy has helped delineate intra-articular pathology and has enabled clinicians to further elucidate the factors responsible for injuries, such as running. The subtle development of degenerative changes may be a result of repetitive impact loading associated with this sport. This study presents a population of runners with common pathologic acetabular changes. TYPE OF STUDY: Case series. METHODS: Eight high-level runners with an average age of 36 years (range, 19 to 45 years) were seen for complaints of increasing hip pain with running without any history of macrotrauma. All of the patients had either run several marathons (4), were triathletes (1), Olympic middle distance runners (1), or had run more than 10 miles per week for longer than 5 years (2). Plain radiographic analysis revealed no degenerative changes and an average center-edge (CE) angle of 36.7 degrees (range, 28 degrees to 44 degrees). RESULTS: All patients underwent hip arthroscopy with labral debridement. In 6 patients (75%), a chondral injury of the acetabular cartilage underlying the labral tear was noted. In addition, 3 patients had ligamentum teres disruptions. CONCLUSIONS: It is possible that the development of these tears is the result of subtle instability, which may be exacerbated by running, eventually leading to labral tearing and possible ligamentum teres disruption. While perhaps concurrently, subtle acetabular dysplasia may play a role. Although this study does not confirm an association between running and the development of labral tears or chondral lesions in the hip, it certainly questions whether there is an injury pattern common to this population, a "runner's hip." LEVEL OF EVIDENCE: Level IV. PMID- 15891726 TI - Arthroscopic partial limbectomy in hip joints with acetabular hypoplasia. AB - PURPOSE: The purpose of this study was to clarify whether arthroscopic partial excision of a torn labrum in acetabular hypoplasia leads to rapid progression of osteoarthritis. TYPE OF STUDY: Clinical case study. MATERIALS AND METHODS: We performed arthroscopic partial limbectomy in 10 hips with pre- or early osteoarthritis secondary to acetabular hypoplasia. All the patients were females with mechanical symptoms such as giving way, catching, locking, or clicking sound. Their ages at the time of surgery ranged from 14 to 62 years (mean, 33.7 y). The follow-up period ranged from 2 to 14 years (mean, 8 y). None of the hips had subluxation. Arthroscopic surgery was performed in the supine position on a traction table, using a simultaneous 3-directional approach. The preoperative center-edge (CE) angle ranged from 0 degrees to 20 degrees (mean, 12.4 degrees). RESULTS: In all patients, preoperative radiographs and magnetic resonance imaging (MRI) depicted no tear of the acetabular labrum, whereas arthroscopic examinations revealed tears incarcerated in the joint space. Coxalgia improved within 4 or 5 days after surgery in all patients, showing that the torn labrum had caused the pain. The tears were classified as bucket-handle type in 6 hips, degenerative type in 3 hips, and flap type in 1 hip. Mechanical symptoms had subsided or disappeared in all hips postoperatively for a mean follow-up of 8 years. Radiographs revealed no evidence of osteoarthritis progression, although one case with a preoperative CE angle of 0 degrees showed lateralization of the femoral head. CONCLUSIONS: Labral tears in acetabular hypoplasia or dysplastic hip are mostly the bucket-handle type, located at the anterosuperior part of the acetabulum. Incarceration of the torn labrum in the joint space causes coxalgia. Rapid progression of osteoarthritis after limbectomy was not observed up to 8 years after surgery in this series. LEVEL OF EVIDENCE: Level IV. PMID- 15891727 TI - Effect of simulated shoulder thermal capsulorrhaphy using radiofrequency energy on glenohumeral fluid temperature. AB - PURPOSE: To determine joint fluid temperatures at different time intervals during treatment with radiofrequency energy (RFE) applied in intermittent and continuous treatment manners under flow or no-flow conditions using a simulated shoulder joint model. TYPE OF STUDY: In vitro measurement of simulated joint fluid temperature during RFE treatment. METHODS: A custom-built jig with a chamber (volume size, 25 mL) was used to mimic the adult human shoulder. Three RFE systems: Vulcan EAS plus TAC-S probe (Smith & Nephew Endoscopy, Andover, MA); VAPR II plus End-Effect Electrode (Mitek, Westwood, MA); and ArthroCare 2000 plus TurboVac 90 degrees probe (ArthroCare, Sunnyvale, CA) were tested in the chamber with saline solution initially set at 23 degrees C. Each RFE probe was applied in a paintbrush pattern on the capsular tissue in the chamber and a fluoroptic thermometry probe was placed 1 cm above the RFE treatment probe to record the fluid temperature. Both intermittent and the continuous treatment manners were tested under flow and no-flow conditions. For each probe/manner/flow combination, 6 bovine capsular tissue specimens were tested (n = 6). All data were recorded using a HyperTerminal software program (Hilgraeve Inc, Monroe, MI) into a personal computer. RESULTS: When using intermittent and continuous treatment manners with flow, all recorded chamber fluid temperatures for all tested RFE probes at each time interval were below 40 degrees C. Under no-flow conditions, with intermittent treatment, the ArthroCare probe caused joint fluid temperatures to exceed 50 degrees C after 70 seconds of RFE treatment. With the continuous treatment, the ArthroCare caused chamber fluid temperatures to exceed 65 degrees C after 2 minutes of treatment. The highest mean recorded chamber fluid temperature was caused by ArthroCare probe, which reached 80 degrees C at 3 minutes. For all probes, continuous treatment caused significantly higher chamber fluid temperatures than intermittent treatment. CONCLUSIONS: The results of this study indicate that using flow during thermal capsulorrhaphy could lower joint fluid temperature to prevent heated joint fluid from killing chondrocytes of articular cartilage, and the intermittent treatment manner caused lower fluid temperature compared with continuous treatment within the RFE-treated shoulder joint. CLINICAL RELEVANCE: Articular cartilage of the humeral head may suffer potential thermal injury from heating of joint fluid during RFE thermal capsulorrhaphy. PMID- 15891728 TI - Open versus arthroscopic rotator cuff repair: a comparative view of 96 cases. AB - PURPOSE: This study was performed to review and compare the outcome of a consecutive series of 96 rotator cuff repairs performed with an open versus arthroscopic technique. The arthroscopic repairs include the beginning of the learning curve. TYPE OF STUDY: Case series. METHODS: In a 24-month period from 1999 to 2001, 95 patients underwent a cuff repair with bony reattachment by 1 surgeon; 4 patients had surgery on both shoulders. We present subjective outcome data from 96 of 99 operated shoulders by the use of a visual analog scale (VAS), the Simple Shoulder Test (SST), and the question of overall patient satisfaction. Thirty shoulders undergoing surgery before February 2000 had open cuff repair (12 classic open, 18 mini-open) and 66 shoulders undergoing surgery after February 2000 had an arthroscopic cuff repair with suture anchors. All but 3 shoulders had the supraspinatus/infraspinatus tendon refixed, 32% had a subscapularis reattachment, and 21% an additional SLAP repair. RESULTS: Groups A (30 shoulders, open repair) and B (66 shoulders, arthroscopic repair) were comparable concerning tear size, cause of the tear, professional shoulder strain, and preoperative pain intensity. At follow-up evaluation (15-40 months after surgery), group B had significantly better pain relief on the VAS (P < .05), more yes answers in the SST (9.7 vs. 8.7, not significant), and a higher patient satisfaction rate of 92.4% versus 80% (not significant). From the 12 questions of the SST, all but 1 showed a higher percentage of yes answers in group B; for 3 questions concerning mobility the difference was significant (P < .07). There was no correlation between tear size and outcome in the SST. CONCLUSIONS: Arthroscopic cuff repair yielded equal or better results than open repair, even at the beginning of the learning curve. Patients with an arthroscopic repair had a significantly better decrease in pain and a better functional result concerning mobility. Arthroscopic repair is successful for large and small tears. Biomechanically, large tears might even benefit more than small ones. LEVEL OF EVIDENCE: Level IV. PMID- 15891729 TI - Immediate postoperative fluid retention and weight gain after shoulder arthroscopy. AB - PURPOSE: The primary purpose of this study was to determine the amount of weight gain immediately after shoulder arthroscopy. In addition, patient and surgical factors correlated with weight gain were analyzed. We hypothesized that there would be a significant increase in weight immediately after shoulder arthroscopy and that the amount of weight gain would correlate with a number of surgical factors. TYPE OF STUDY: Observational case series. METHODS: Fifty-three patients were enrolled in the study. There were 34 male and 19 female patients, with a mean age of 47.1 +/- 13.8 years. All patients were weighed before and after surgery using the same weight scale by the same observer. Weight gain was calculated and adjusted to account for the weight of the dressing and sling. Weight gain then was correlated to various clinical and surgical variables. Pearson correlation coefficients, Student t tests, and stepwise regression were used to determine significant correlations between clinical and surgical variables and weight gain. RESULTS: The mean amount of net weight gain was 8.7 +/ 3.9 lb (range, 0.8-18.8 lb), representing 4.6% +/- 2.1% of preoperative weight. The mean amount of weight gain attributable to arthroscopy fluid only was 4.2 +/- 3.8 lb (range, 0-14.5 lb), representing a gain of 2.2% +/- 2.0% of preoperative weight. The mean amount of intravenous fluid infused was 1,885 +/- 547 mL, and the mean amount of normal saline arthroscopy fluid used was 30 +/- 24 L. Surgical time, the amount of arthroscopy fluid, the size of the rotator cuff tear, the number of tendons involved, the presence of a subscapularis tear, the number of procedures performed, the concomitant performance of a subacromial decompression, the number of BioCorkscrew (Arthrex, Inc., Naples, FL) anchors used, and the total number of anchors used all correlated with increasing weight gain (all P < .05). A procedure of stepwise regression selection did not identify any quantitative parameters attributable to weight gain other than the earlier described parameters. There were no significant intraoperative or postoperative complications attributable to the amount of weight gain. CONCLUSIONS: Weight gain immediately after shoulder arthroscopy is a common finding. Although no complications were seen in this group of patients, both patients and surgeons should be aware of this concern after shoulder arthroscopy and the potential complications related to it. PMID- 15891730 TI - Biomechanical evaluation of different suture anchors for the stabilization of anterior labrum lesions. AB - PURPOSE: The aim of the study was to investigate the biomechanical performance of different devices for anterior shoulder reconstruction in a clinically relevant human cadaver shoulder model. TYPE OF STUDY: Biomechanical cadaveric study. METHODS: The soft tissue, except for the joint capsule, was dissected from 28 fresh-frozen human shoulder specimens. A Bankart lesion was created and repaired with 3 suture anchors. The devices tested were the 2.4-mm Fastak (Arthrex, Karlsfeld, Germany) with either Ethibond (Ethicon, Westwood, MA) no. 2 or 3, the 2.8-mm Fastak with Ethibond no. 2, the Panalok (Mitek, Norwood, MA) with Ethibond no. 2 with a modified setting technique, and the 8-mm Suretac (Acufex Microsurgical, Mansfield, MA). The specimens were anteriorly dislocated in 60 degrees glenohumeral abduction and 90 degrees external rotation. Load was measured continuously and failures were noted. The experimental groups were compared with each other and with a control group with intact shoulders. RESULTS: Bankart repair with 2.4-mm Fastaks and Ethibond no. 2 and 3 failed at a mean load of 342.0 N and 692.2 N, respectively, predominantly by suture rupture at the eyelet of the anchor. Shoulder reconstruction with 2.8-mm Fastaks failed at an average load of 722.8 N, mostly by suture rupture at the knot. The difference to the 2.4-mm Fastak with Ethibond no. 2 was significant (P < .05). The mean load to failure was 983.8 N in the Panalok group, and the most frequent failure mode was suture rupture at the knot. Bankart repair with Suretac failed at an average load of 468.4 N, typically by capsular damage at the repair site. Capsular injuries in the control group occurred at a mean load of 958.2 N. Anterior shoulder reconstruction with 2.4-mm Fastak with Ethibond no. 2 and with Suretac were statistically significantly inferior to the control group (P < .05). CONCLUSIONS: All tested devices allow stable anterior shoulder reconstruction, but we recommend the Panalok and the 2.8-mm Fastak because they provided greater stability than either the 2.4-mm Fastak or the Suretac in our experiments. Suretac can possibly cause additional capsular damage in redislocations after Bankart repair. CLINICAL RELEVANCE: This cadaveric study gives an insight into the biomechanical performance of a Bankart repair with different devices directly postoperative and shows possible failure modes and additional injuries in case of early traumatic redislocation. PMID- 15891731 TI - Results of arthroscopic reduction and percutaneous fixation for acute displaced scaphoid fractures. AB - PURPOSE: This study used percutaneous techniques augmented by simultaneous wrist arthroscopy to visualize the fracture and thus confirm the fracture alignment and reduction and also to assesses the concurrent associated ligament injuries. TYPE OF STUDY: Retrospective study. METHODS: Arthroscopy was used to help to reduce scaphoid fractures and assess soft-tissue injuries in 15 acute cases (13 male and 2 female patients). The fractures were treated by reduction under arthroscopic control and percutaneous fixation with the cannulated interosseous compression screw. Soft-tissue lesions were also treated at the same time using debridement, suture repair, or K-wire transfixation. The average age of the patients was 29.2 years (range, 19 to 48 years). RESULTS: Two patients (13.3%) had scapholunate (SL) ligament injuries, and both exhibited partial tear of the SL ligament. Four patients (26.7%) suffered lunotriquetral (LT) ligament injuries and received ligament debridement, K-wire fixation of the LT joint, and splinting. Six patients (40%) had chondral fractures. Additionally, the triangular fibrocartilage complex (TFCC) was torn in 5 patients (33%). Finally, 5 patients (33%) suffered radioscaphocapitate ligament or long radiolunate ligament injuries. All fractures healed without malunion or nonunion and, at follow-up of 24 to 28 months, 11 patients had excellent results and 4 had good results based on Mayo Modified Wrist Scores. CONCLUSIONS: We believe that arthroscopic reduction may be considered for scaphoid fractures because this approach can use a single procedure to achieve acceptable restoration of fractures as well as assessment and management of soft-tissue lesions. LEVEL OF EVIDENCE: Level IV Therapeutic Study, case series with no, or historical, control group. PMID- 15891732 TI - Arthroscopic patellar "bankart" repair after acute dislocation. AB - We present a case of acute patellar dislocation in a skeletally immature patient treated with arthroscopic medial patellofemoral ligamentous complex repair using suture anchors with a horizontal mattress suture technique. Patellar dislocation is a common problem in the skeletally immature. Treatment is controversial for first-time dislocators because of the high rate of recurrent instability and functional disability in these patients. Surgical repair of the medial restraints may decrease recurrent instability, although anterior knee pain and crepitus is a common finding after open surgical techniques. Arthroscopic medial retinacular imbrication has been described for patients with persistent instability with good short-term results. Acute dislocations may cause avulsion of the medial patellofemoral ligamentous complex from the patella; this is amenable to direct primary repair to prevent recurrent instability and avoid the morbidity of open surgery. This technique recreates normal anatomy and function with minimally invasive surgery. PMID- 15891733 TI - Locked lateral patella dislocation with generalized ligamentous laxity after arthroscopic lateral release of the knee. AB - We describe the case of a 14-year-old boy with patellar instability on both sides resulting from ligamentous hyperlaxity and dysplasia of the lateral femoral condyle who had previously undergone an arthroscopic lateral release as well as plication of the medial capsule. The patient presented to our clinic 2 years after surgery with a locked lateral patella dislocation on the left side. The surgical correction involved a substantial open lateral release including an excision of the scar tissue and stabilization of the patella in the patellofemoral groove by tibial tubercle medialization and plication of the medial capsule. Six months after surgery, the patient achieved a Lysholm score of 90 points and clinical examination indicated a stable knee with a centralized patella without any evidence of subluxation or dislocation. Open lateral release with partial resection of the lateral retinaculum, medial reconstruction, and tibial tubercle osteotomy was the procedure of choice in this patient with habitual patella dislocation caused by generalized ligamentous laxity. PMID- 15891734 TI - Symptomatic anomalous insertion of the medial meniscus. AB - We report a case of a young athlete with a symptomatic anomaly of the medial meniscus. An anomalous portion in conjunction with the anterior horn of the medial meniscus extended to the intercondylar notch of the femur through the surface of the anterior cruciate ligament. This anomalous band was arthroscopically resected and the symptoms completely disappeared. Histologic examination showed fibrocartilaginous tissue compatible with meniscus. PMID- 15891735 TI - Bony lesion recurrence after mosaicplasty for osteochondritis dissecans of the talus. AB - Autogenous osteochondral grafts have recently become popular for use in small, isolated, contained articular cartilage defects. We treated a 26-year-old man who had a cartilage defect measuring 10 x 20 mm in the anteromedial area of the right talus. We performed multiple osteochondral grafting of the lesion with medial malleolar osteotomy from a donor site in the ipsilateral knee joint. Two years after the operation, the patient's ankle pain recurred and the bony lesion in the talus also became osteolytic. Because we believed that only the cartilaginous portions of the osteochondral plugs grafted 2 years previously were fully fixed and viable, and that recurrence had occurred at the bony portions, at reoperation we performed curettage of the bony lesions and grafted iliac bone into the lesions with fenestration of the inferomedial ankle joint cartilage, not grafted plug cartilage. Therefore, probably because of overuse, the bony lesion in the talus had recurred 2 years after the first operation, but the grafted hyaline cartilage had survived. Autogenous osteochondral grafting into the talus, unlike the knee joint, should be done with care to ensure there is no sclerotic bone surrounding the lesion in patients with long-standing symptoms and recurrence of bony lesions. PMID- 15891736 TI - Neuropraxia of the cutaneous nerve of the cervical plexus after shoulder arthroscopy. AB - This article presents uncommon cases of neuropraxia of the lesser occipital nerve and the greater auricular nerve after arthroscopic surgery of the shoulder in the beach-chair position under general anesthesia. The lesser occipital nerve and the greater auricular nerve are superficial ascending branches of the cervical plexus. These 2 superficial nerves may be easily vulnerable because of their superficial anatomic locations. We assumed that the severity of the neuropraxia of superficial branches of the cervical plexus was related to the degree of rotation and deviation of the head and neck, the duration of the procedure, and compression by head strap and elastic bandage used for fixing the head to the rectangular-shaped headrest of the beach-chair device. We recommend that during surgery in the beach-chair position, the auricle be protected and covered with cotton and gauze to avoid direct compression and the position of the head and neck be checked and corrected frequently. We hope for a new design of the headrest of the beach-chair device to prevent neuropraxia and to attach the head firmly and safely. PMID- 15891737 TI - Arthroscopic repair of a humeral avulsion of the glenohumeral ligament lesion. AB - We describe 3 cases of an all-arthroscopic technique for repair of a humeral avulsion of the glenohumeral ligament (HAGL) lesion and the postoperative clinical outcomes. From a technical perspective, the most critical part of the surgeries was the anchor insertion at an optimal position on the humerus in order to achieve proper tension of the glenohumeral ligament. The arm-free beach-chair position, which facilitates maximum internal rotation, use of a 70 degrees angled arthroscope, and an anterior-inferior trans-subscapularis tendon portal were considered key factors to accomplish this procedure. PMID- 15891738 TI - Arthroscopic technique for treatment of synovial chondromatosis of the glenohumeral joint. AB - Synovial chondromatosis of the shoulder has been treated in the past with an open arthrotomy, removal of loose bodies, and synovectomy. Several authors have described arthroscopy as a means of treating this rare disorder because of the excellent visualization of the glenohumeral joint, decreased morbidity, and early rehabilitation potential. This article offers a technique in which a large-bore cannula and high pressure and flow are used to facilitate the removal of loose bodies from the joint. PMID- 15891739 TI - The arthroscopic active compression test. AB - Pathology of the biceps tendon is often a factor in the pathogenesis of shoulder pain. However, diagnosis and management of such pathology remains controversial. Multiple tests have been described to identify pathology related to the biceps tendon and biceps anchor. The active compression test is one such test that has been used clinically to identify patients with superior labral/biceps anchor pathology. An arthroscopic version of this test is described to identify biceps pathology intraoperatively and to aid the surgeon in making diagnostic and treatment decisions. PMID- 15891740 TI - Arthroscopic repair of glenoid fractures using suture anchors. AB - Anterior glenoid fractures associated with glenohumeral instability are normally treated with open fixation of the fragment. However, recent technical development enables this procedure to be performed under arthroscopic control using screws or even suture anchors if the fragment is small enough. We developed a unique method of stabilizing a larger fragment with suture anchors and the use of a clear imaging method of three-dimensional computed tomography (3D-CT) with the humeral head eliminated, preoperatively and postoperatively. In the present article, we report a detailed method of arthroscopic stabilization for a consecutive series of 8 patients, with an average of 27% bone loss (fragment size) against the lower part of the circular glenoid portion, using suture anchors. PMID- 15891741 TI - Arthroscopic thermal capsulorrhaphy as treatment for the unstable paralytic shoulder. AB - Patients who present with global capsular laxity and glenohumeral instability frequently can be treated successfully with shoulder girdle musculature strengthening exercises and activity modification. When such injury is caused by a paralytic shoulder, a rehabilitation program may not be a viable treatment option. Presented in this article are 3 patients with global capsular laxity and glenohumeral instability as a result of shoulder paralysis. We also describe an arthroscopic technique of thermal capsulorrhaphy. In our experience, arthroscopic thermal capsulorrhaphy used to treat global capsular laxity and glenohumeral instability resulting from a paralytic shoulder has decreased symptoms of shoulder instability and has significantly reduced shoulder pain. PMID- 15891742 TI - Technique to facilitate graft passage in posterolateral reconstructions of the knee. AB - This report describes a novel technique to facilitate passage of the transtibial limb of grafts used for reconstruction of the posterolateral corner of the knee. This new method, which places and passes the graft within a 24F (8-mm) chest tube, eliminates the problem of graft impingement at the opening of the posterior tibial tunnel and the need to enlarge the tunnel to achieve graft passage. Enlarging the tunnel is detrimental to obtaining the snug fit that is necessary to maximize incorporation of the graft and minimize postoperative tunnel expansion. This chest tube technique, which has been used in 20 cases to date, makes graft passage expeditious and requires no special instrumentation. PMID- 15891743 TI - Arthroscopic treatment of popliteal cyst and visualization of its cavity through the posterior portal of the knee. AB - In regard to arthroscopic treatment of popliteal cysts, we explored the refuted pathology for popliteal cysts proposed by others. Here we introduce an arthroscopic technique using posterior portals to treat a popliteal cyst based on our observation that the opening of the cyst in the joint is a slit-like structure in the posterior wall of the capsule. By disrupting this slit-like structure with our procedure, the popliteal cyst ceased to be palpable and was no longer symptomatic. This technique also provides excellent arthroscopic visualization of the cavity of the popliteal cyst through the knee joint approach. After completion of the resection of the opening, we can easily insert an arthroscope into the cavity of the popliteal cyst from the posteromedial portal through the resected opening. Arthroscopic visualization of the cavity of the cyst showed that the inside wall of the cavity was smooth and had no synovitis. We believe that to disrupt this slit structure is the most pathologically reasonable procedure to treat popliteal cysts surgically. PMID- 15891744 TI - Anatomic reconstruction of the anteromedial and posterolateral bundles of the anterior cruciate ligament using hamstring tendon grafts. PMID- 15891746 TI - Abdominal aortic aneurysm screening during transthoracic echocardiography in an unselected population. AB - OBJECTIVE: We sought to investigate the echocardiographic prevalence of abdominal aortic aneurysm (AAA) in an unselected group of patients referred for regular transthoracic echocardiography (TTE). METHODS: Prospectively, during a 3-month period, a limited ultrasound examination of the infrarenal aorta was performed. AAA was defined as a diameter of 30 mm or more. RESULTS: The abdominal aorta could be visualized in 742 patients. The prevalence of AAA was 4.6%. AAA prevalence increased with age, especially in men. In 34 patients AAA was unknown and aortic diameters exceeded 50 mm in 4 patients. Two underwent elective but urgent operation. Patients with AAA were older and had an increased ascending aorta diameter, larger left ventricular dimensions, higher left ventricular mass index, and lower ejection fraction. CONCLUSION: AAA is prevalent in patients referred for regular TTE. Routine rapid screening of the abdominal aorta during TTE is beneficial and should, therefore, be part of a standard TTE examination for patients older then 50 years. PMID- 15891747 TI - Residents can be trained to detect abdominal aortic aneurysms using personal ultrasound imagers: a pilot study. AB - Our objective was to test the hypothesis that internal medicine residents can be trained to screen for abdominal aortic aneurysm (AAA) using personal ultrasound imagers. We trained 5 randomly chosen internal medicine residents to image the abdominal aorta for patients with risk factors for AAAs using personal ultrasound imagers. Residents were trained in 3 or 4 one-on-one sessions with an instructor. To be eligible, patients had to be older than 65 years and have hypertension. After training, each of the 5 residents studied 3 patients independently. In 12 of the residents' 15 unsupervised studies, their abdominal aorta measurements were within 5 mm of the instructor's measurements with standard echocardiography (mean difference 3 mm, range 0-6 mm). Residents detected 3 previously unknown AAAs measuring 5.2, 4.2, and 3.9 cm in diameter. We conclude residents can be trained to image the abdominal aorta with personal ultrasound imagers and to identify AAAs in patients at risk. PMID- 15891748 TI - Strain rate imaging is a superior method for the assessment of regional myocardial function compared with Doppler tissue imaging: a study on patients with transcatheter device closure of atrial septal defect. AB - To compare the value of strain and strain rate imaging in assessing regional myocardial function to that of myocardial velocities, we studied 10 patients late after successful percutaneous atrial septal defect closure. Analysis was performed for atrial longitudinal peak systolic velocity, strain, and strain rate from the midsegment of interatrial septum, in correspondence of the device, and from the lateral left atrial wall. Placing the sample volume in the middle part of the atrial septal defect occluder, a bulky noncontractile element, passively moved by global heart motion, the new ultrasonic-derived myocardial deformation indexes demonstrated almost the absence of any deformations, whereas myocardial velocities failed to significantly discriminate between this noncontracting structure and the normal atrial wall. PMID- 15891749 TI - Recovery of stunned myocardium in acute myocardial infarction quantified by strain rate imaging: a clinical study. AB - BACKGROUND: Strain rate (SR) imaging (SRI) is a tissue Doppler-based method of regional myocardial deformation imaging. The aim of this study was to see whether SRI could quantify changes in myocardial mechanical function after an acute myocardial infarction, and to follow the time course of these changes. METHODS: In all, 26 consecutive patients with first-time acute myocardial infarctions were examined on days 1, 7, and 90. Segments were analyzed with SRI and wall-motion score. RESULTS: Peak systolic SR in infarcted segments increased significantly in magnitude from day 1 to 7 (-0.45 to -0.68 s -1 , P < .001), but not after day 7. The deformation rate in border zone segments also increased significantly from day 1 to 7 (-0.8 to -0.95 s -1 , P < .05), with no further significant changes at 3 months. In normal segments, peak systolic SR decreased in magnitude during the first week. Systolic strain showed similar results as peak systolic SR. CONCLUSION: SRI can demonstrate small changes in deformation rate from midinfarct through the infarct and border zone to normal myocardium. It can also show changes over time, probably as a result of recovery of stunned myocardium. PMID- 15891750 TI - Automated analysis of strain rate and strain: feasibility and clinical implications. AB - BACKGROUND: This study evaluated 3 new automated methods, based on a combination of speckle tracking and tissue Doppler, for the analysis of strain rate (SR) and strain. Feasibility and values for peak systolic strain rate (SR s ) and end systolic strain (S es ) were assessed. METHODS: Thirty patients with myocardial infarction and 30 normal subjects were examined. Customized software with automatic definition of segments was used for automated measurements. SR s and SR es were measured over each segment simultaneously and identified automatically. The study compared tissue Doppler-based SR and strain measurements without (method 1) and with segment tracking (method 2) to speckle tracking-based measurements (method 3). For tracking, speckle tracking and tissue Doppler were used in combination. Standard manual analysis was used as a reference. RESULTS: The automated analysis (16 segments, 3 apical views) required 2 minutes; manual analysis took 11 minutes. Accuracy was compared in 56 segments (28 mid-infarcted and 28 normal) from 28 patients and was 93.9% for method 1, 93.8% for method 2, 95.8% for method 3, and 96.2% for the manual method. In the normal group, mean SR s (0.27 s -1 ) was less with method 3 than with the other methods ( P < .001). CONCLUSIONS: Our findings indicate that automated analysis of SR and strain, with some manual adjustment, is feasible and quicker than manual analysis. Diagnostic accuracy was similar with all methods. SR s was lower in the speckle tracking based method than in the Doppler-based methods. PMID- 15891751 TI - Echocardiography to measure fitness of elite runners. AB - We evaluated the echocardiograms of 134 male and 54 female elite runners who competed over different distances to observe cardiac enlargement and differences among athletes engaged in different types of training, and to relate the echographic dimension with running performance. In male runners, left ventricular (LV) internal diameter at end diastole (LVIDd), LV mass, and left atrial diameter were increased by the length of distance ( P < .001) whereas right ventricular diameter was increased only in marathon runners. In female runners, this association was found when LVIDd, LV mass, and left atrial diameter were adjusted for body surface area ( P < .001). Wall thickness was not related to distance running. Abnormal LV dilation was associated with a low LV ejection fraction for the higher values of LVIDd, but not for the higher indexed LVIDd values. In male runners, LVIDd was associated with 100 m ( r = -0.67, P < .01), 10,000 m ( r = 0.70, P < .001), and marathon ( r = -0.54, P < .001) and in female runners with a 400-m ( r = -0.84, P < .001) race time. These results show that sprint- and endurance-trained runners had an eccentric hypertrophy and that it is necessary to establish a new upper limit of abnormal cavity dilatation as indexed LVIDd. The normal systolic function and the relationship between LVIDd and performance indicate that LV enlargement in elite runners is a physiologic adaptation and that echocardiographic assessment can be used to determine the fitness of runners. PMID- 15891752 TI - Estimation of right ventricular mass by two-dimensional echocardiography. AB - BACKGROUND: This report describes two original echocardiographic approaches to measure right ventricular (RV) mass (RVM). METHODS: In the bullet formula (5/24 pi D1 D2 L), where D1 and D2 are short axes and L the log axis, the RVM is obtained by subtracting the cavity volume from the RV total volume and subsequently multiplying the difference by myocardium density. The second method uses 3 endocardium segments measured at: (1) short axis plane of the aortic valve and left atrium (b1); (2) short axis plane at the midpoint between the tricuspid valve annulus and the apex (b2); and (3) 4-chamber view (h). Those segment lengths are applying in the formula A = [(b1 + b2)/2] x h. The result is multiplied by the wall thickness and by myocardium density. RESULTS: Both formulas were primarily tested in 30 mongrel dogs and have shown good correlation with the true mass ( r = 0.869 with the segments formula and r = 0.819 with the bullet formula). The same method was used in 20 human patients before heart transplant with similar results ( r = 0.810 with the segments formula and r = 0.836 with the bullet formula). CONCLUSIONS: The RVM can be satisfactorily estimated by 2-dimensional echocardiography. The linear regression between the calculated mass (using the smoothest and thinner myocardium thickness) and the actual mass may provide the correction factor for the RVM calculation. Two echocardiographic methods were used to measure right ventricular mass. One of them used a bullet formula variant (5/24 pi D1 D2 L). The second method used 3 endocardium segments measured in 3 2-dimensional echocardiographic planes (short axis of aortic valve and left ventricle, and 4-chamber view), and applied in the formula A = [(b1 + b2)/2] x h. Both formulas have shown good correlation with the true mass in 30 mongrel dogs ( r = 0.869 with the segments formula and r = 0.819 with the bullet formula) and in 20 human patients before heart transplant ( r = 0.810 and r = 0.836, respectively). PMID- 15891753 TI - Prognostic importance of various echocardiographic right ventricular functional parameters in patients with symptomatic heart failure. AB - Little is known about the prognostic importance of right ventricular (RV) systolic and diastolic function. The purpose of this study was to determine the prognostic power of systolic and diastolic RV functional parameters derived from Doppler tissue imaging of tricuspid annular motion and to assess whether their combination might improve the risk stratification of patients with heart failure. In all, 140 patients with symptomatic heart failure and left ventricular ejection fraction of 40% or less underwent standard echocardiography, Doppler tissue imaging of tricuspid annular motion, and right heart catheterization. They were followed up for a mean period of 17 months for cardiac-related death and nonfatal cardiac events including the implantation of cardioverter-defibrillator and hospitalization for heart failure decompensation. A total of 48 cardiac events occurred; 19 patients died, 26 were hospitalized for heart failure decompensation, and 3 because of the need for implantation of a cardioverter defibrillator. The peak tricuspid annular velocity during systolic ejection of 10.8 cm/s or less, peak early diastolic tricuspid annular velocity of 8.9 cm/s or less, tricuspid annular acceleration during isovolumic contraction of 2.52 m/s 2 or less, and Doppler RV index (Tei index) of 1.20 or more were found to significantly worsen survival or event-free survival. However, their combination significantly exceeded the predictive potential of individual parameters. The worst survival was predicted by the combination of peak tricuspid annular velocity during systolic ejection of 10.8 cm/s or less plus peak early diastolic tricuspid annular velocity of 8.9 cm/s or less plus tricuspid annular acceleration during isovolumic contraction of 2.52 m/s 2 or less (relative risk 6.17, P < .001), whereas the worst event-free survival was identified by the combination of peak tricuspid annular velocity during systolic ejection of 10.8 cm/s or less plus peak early diastolic tricuspid annular velocity of 8.9 cm/s or less plus Doppler RV index (Tei index) of 1.20 or more (relative risk 3.62, P < .001). In conclusion, the combination of RV systolic and diastolic functional parameters represents a very powerful tool for risk stratification of patients with symptomatic heart failure. PMID- 15891754 TI - Abnormal septal motion affects early diastolic velocities at the septal and lateral mitral annulus, and impacts on estimation of the pulmonary capillary wedge pressure. AB - Abnormal motion of the interventricular septum (ASM), seen post cardiac operation, with left bundle branch block or right ventricular pacing, may affect septal mitral annular motion and correlation of the ratio between the velocity of early diastolic mitral inflow and the early diastolic mitral annular velocity (E/Ea) with pulmonary capillary wedge pressure (PCWP). We examined the effect of ASM on the relationship between E/Ea and E/Vp (propagation velocity of mitral inflow) ratios and PCWP in adult patients in the intensive care unit (14 with normal septal motion [NSM], 36 with ASM) undergoing echocardiography and pulmonary artery catheterization. E/Ea correlated well with PCWP during NSM ( r = 0.86 lateral annulus, r = 0.75 septal annulus), but poorly during ASM ( r = 0.36 lateral annulus, r = 0.39 septal annulus). E/Vp correlated poorly with PCWP ( r = 0.05 NSM, r = 0.17 ASM). For patients who are critically ill, E/Vp ratios poorly estimate PCWP. During NSM, E/Ea ratios measured at the lateral or septal annulus correlate well with PCWP. ASM affects E/Ea ratios at both the septal and lateral annulus, making E/Ea ratios unreliable for estimating PCWP in this group. PMID- 15891755 TI - Myocardial performance index in female rats with myocardial infarction: relationship with ventricular function parameters by Doppler echocardiography. AB - BACKGROUND: The aim of the study was to analyze the myocardial performance index (MPI), its relationship with the standard variables of systolic and diastolic functions, and the influence of time intervals in an experimental model of female rats with myocardial infarction (MI). METHODS: Forty-one Wistar female rats were submitted to surgery to induce MI. Six weeks later, Doppler echocardiography was performed to assess infarct size (IS,%), fractional area change (FAC,%), ejection fraction biplane Simpson (EF), E/A ratio of mitral inflow, MPI and its time intervals: isovolumetric contraction (IVCT, ms) and relaxation (IVRT, ms) times, and ejection time (ET, ms); MPI = IVCT + IVRT/ET. RESULTS: EF and FAC were progressively lower in rats with small, medium and large-size MI ( P < .001). E/A ratio was higher only in rats with large-size MI (6.25 +/- 2.69; P < .001). MPI was not different between control rats and small-size MI (0.37 +/- 0.03 vs 0.34 +/- 0.06, P = .87), but different between large and medium-size MI (0.69 +/- 0.08 vs 0.47 +/- 0.07; P < .001) and between these two compared to small-size MI. MPI correlated with IS (r = 0.85; P < .001), EF (r = -0.86; P < .001), FAC (r = 0.77; P < .001) and E/A ratio (r = 0.77; P < .001, non-linear). IVCT was longer in large size MI compared to medium-size MI (31.87 +/- 7.99 vs 15.92 +/- 5.88; P < .001) and correlated with IS (r = 0.85; P < .001) and MPI (r = 0.92; P < .001). ET was shorter only in large-size MI (81.07 +/- 7.23; P < .001), and correlated with IS (r = -0.70; P < .001) and MPI (r = -0.85; P < .001). IVRT was shorter only in large-size compared to medium-size MI (24.40 +/- 5.38 vs 29.69 +/- 5.92; P < .037), had borderline correlation with MPI (r = 0.34; P = .0534) and no correlation with IS (r = 0.26; p = 0.144). CONCLUSIONS: The MPI increased with IS, correlated inversely with systolic function parameters and had a non-linear relationship with diastolic function. These changes were due to the increase of IVCT and a decrease of ET, without significant influence of IVRT. PMID- 15891756 TI - Noninvasive analysis of myocardial function using high-resolution Doppler tissue echocardiography in rats. PMID- 15891757 TI - Development of a new animal model of chronic mitral regurgitation in rats under transesophageal echocardiographic guidance. AB - Large animal models (dog and sheep) are often used for the investigation of the pathophysiology of chronic mitral regurgitation (MR). A major limitation of large animal models is cost. The aim of this study was to develop a new animal model of chronic MR. Left thoracotomy was performed in 34 rats. Under the guidance of transesophageal echocardiography, a fine needle was inserted into the left ventricle (LV) to damage the mitral leaflets and produce MR. Serial transthoracic echocardiography was performed to assess LV remodeling and function. Left atrial and LV diameters were significantly larger, and LV fractional shortening was lower in the MR group than in the sham group. The 150-day survival was 59% in the MR group and 100% in the sham group (P < .01). This new animal model of chronic MR may be used in the study of the pathophysiology of chronic MR and pharmacologic therapies. PMID- 15891758 TI - Contrast echocardiography improves the diagnostic yield of transthoracic studies performed in the intensive care setting by novice sonographers. AB - BACKGROUND: Bedside portable echocardiography in the intensive care department (ICU) is technically difficult, but crucial for directing patient care. Prior studies have shown contrast echocardiography (CE) in the ICU clarifies left ventricular wall motion when performed by experienced sonographers (ESO). However, in most hospitals, ESO are unavailable around the clock, and less experienced cardiovascular fellows or trainees may be asked to perform these examinations. METHODS: Transthoracic echocardiograms were retrospectively evaluated by level III trained echocardiographers for 213 patients in the ICU. Most were performed to assess left ventricular function (65% or 139 of 213) and were scanned by cardiology fellows (70% or 149 of 213) with less than 3 months echocardiography experience. Contrast agent was used in 29% (62 of 213) of all patients. RESULTS: The conversion of suboptimal or diagnostically inadequate apical 4- and 2-chamber views to diagnostically adequate with contrast was statistically significant when performed by both cardiology fellows and ESO (Fischer exact test, P < .0002). CONCLUSIONS: CE is effective in improving the diagnostic yield of transthoracic echocardiographic ICU studies performed by both novice sonographers and ESO. Using cardiology fellows to perform CE in this setting can be appropriate, particularly in after-hour situations, when ESO are not always available and the clinical question is left ventricular function. Results also suggest cardiology fellows can easily learn CE. PMID- 15891759 TI - Conservative therapeutic approach in a case of coronary dissection extending into the aortic root complicating coronary angioplasty. PMID- 15891760 TI - A case report of absent right superior vena cava with persistent left superior vena cava: echocardiographic findings. PMID- 15891761 TI - Coronary artery vasospasm during dobutamine stress echocardiography. AB - Compared with the frequent occurrence of S-T segment depression, the development of S-T segment elevation during dobutamine stress echocardiography is a relatively rare finding, especially in the setting of nonobstructive coronary artery disease. We present two patients who developed chest discomfort with S-T segment elevation during dobutamine stress testing. Both patients had unremarkable baseline echocardiograms and no history suggestive of coronary vasospasm. Subsequent coronary angiography revealed nonobstructive disease. It is speculated that the likely presence of endothelial dysfunction coupled with mild coronary atherosclerosis caused an imbalance between the coronary vasodilatory and vasoconstrictive forces. The inhibition of the vasodilatory effects of beta adrenergic receptor stimulation and flow-mediated dilation of dobutamine was overcome by the vasoconstrictive effects of dobutamine-stimulated alpha-1 receptor activation; thus, contributing to the development of coronary vasospasm. This condition is important to recognize and appreciate as the initial treatment for this condition favors the use of nitrates over beta-blockade. PMID- 15891762 TI - Evaluation of right pulmonary artery obstruction by echocardiography in a 32-year old woman with suggested histoplasmosis. PMID- 15891763 TI - Vegetation and bilateral congenital coronary artery fistulas. AB - This report describes the case of a man who presented with fever, weight loss, and Streptococcus mitis septicemia. He was found to have congenital bilateral coronary artery fistula, a rare condition. There was no evidence of vegetation on the heart valves on transthoracic or transesophageal echocardiography. Instead, transesophageal echocardiography showed vegetation within the coronary sinus near its entrance into the right atrium. Coronary angiography confirmed the presence of a left circumflex artery to coronary sinus fistula and a right coronary artery to coronary sinus fistula. To the authors' knowledge this combination of findings has never been reported before. PMID- 15891764 TI - Accessory mitral valve in an adult population: the role of echocardiography in diagnosis and management. AB - Accessory mitral valve is a rare congenital abnormality and is an unusual cause for subvalvular left ventricular outflow tract (LVOT) obstruction. It is detected first in children and is very rarely noticed in adults. The most common clinical presentation is symptomatic LVOT obstruction. We present a case series of 5 adult patients with varying clinical presentations in which the accessory mitral valve was diagnosed using echocardiography. Three patients presented with varying degrees of symptomatic LVOT obstruction, one presented with recurrent transient ischemic attack and stroke, and one patient was incidentally diagnosed during echocardiography to exclude endocarditis. Accessory mitral valve should be suggested in patients with LVOT obstruction. PMID- 15891765 TI - Development and testing of a computerised 24-h recall questionnaire measuring fruit and snack consumption among 9-11 year olds. AB - OBJECTIVE: To evaluate the validity, reliability and sensitivity of a computerised single day 24-h recall questionnaire designed for the comparison of children's fruit and snack consumption at the group (school) level. DESIGN: Relative validity and reliability were assessed in relation to (i) intake at school and (ii) intake throughout the whole day, using diary-assisted 24-h recall interviews and a 7-day test-retest procedure. Sensitivity was assessed in relation to intake by comparing results from schools with differing food policies, and by sex. SUBJECTS AND SETTING: Eight schools took part in the validity and reliability assessments, with 78 children completing the 24-h recall interviews and 195 children completing the test-retest procedure. A total of 43 schools (1890 children) took part in the sensitivity analysis. All children were aged 9-11 y. All schools were in South Wales and South-west England. RESULTS: For fruit intake at school, the questionnaire showed fair levels of validity at the individual level (kappa = 0.29). At the group level, there were little or no differences in fruit intake at school between the two measures and two occasions. The questionnaire was sufficiently sensitive to identify statistically significant differences between girls and boys, and between schools with different food policies. For snack intake at school, validity at the individual level was slightly lower (kappa = 0.220.25), but the data remained of value in analyses at the group level. For fruit and snack intake throughout the whole day there was little agreement at the individual level (kappa = 0.00-0.06), and at the group level there tended to be substantial differences between the two measures and two occasions. CONCLUSIONS: The computerised questionnaire is a quick and cost-effective means of assessing children's consumption of fruit at school. While further development is required to improve validity and reliability, it has the potential to be particularly useful in randomised controlled trials of school-based dietary interventions. PMID- 15891766 TI - Roles for replichores and macrodomains in segregation of the Escherichia coli chromosome. AB - Recent work has highlighted two main levels of global organization of the Escherichia coli chromosome. Macrodomains are large domains inferred from structural data consisting of loci showing the same intracellular positioning. Replichores, defined by base composition skews, coincide with the replication arms in normal cells. We used chromosome inversions to show that the dif site, which resolves chromosome dimers, only functions when located at the junction of the replichores, whatever their size. This is the first evidence that replichore polarization has a role in chromosome segregation. We also show that disruption of the Ter macrodomain provokes a cell-cycle defect independent from dimer resolution. This confirms the existence of the Ter macrodomain and suggests a role in chromosome dynamics. PMID- 15891767 TI - Mismatch repair participates in error-free processing of DNA interstrand crosslinks in human cells. AB - DNA interstrand crosslinks (ICLs) present formidable blocks to DNA metabolic processes and must be repaired for cell survival. ICLs are induced in DNA by intercalating compounds such as the widely used therapeutic agent psoralen. In bacteria, both nucleotide excision repair (NER) and homologous recombination are required for the repair of ICLs. The processing of ICLs in mammalian cells is not clearly understood. However, it is known that processing can occur by NER, which for psoralen ICLs can be an error-generating process conducive to mutagenesis. We show here that another repair pathway, mismatch repair (MMR), is also involved in eliminating psoralen ICLs in human cells. MMR deficiency renders cells hypersensitive to psoralen ICLs without diminishing their mutagenic potential, suggesting that MMR does not contribute to error-generating repair, and that MMR may represent a relatively error-free mechanism for processing these lesions in human cells. Thus, enhancement of MMR relative to NER may reduce the mutagenesis caused by DNA ICLs in humans. PMID- 15891768 TI - Functional regulation of oestrogen receptor pathway by the dynein light chain 1. AB - Overexpression and phosphorylation of dynein light chain 1 (DLC1) have been shown to promote the growth of breast cancer cells. However, the role of DLC1 in the action of the oestrogen receptor (ER) remains unknown. Here, we found that oestrogen induces the transcription and expression of DLC1. DLC1 facilitated oestrogen-induced ER transactivation and anchorage-independent growth of breast cancer cells. We show that DLC1 interacts with ER, and such interaction is required for the transactivation-promoting activity of DLC1. Further, DLC1 expression led to enhanced recruitment of the DLC1-ER complex to the ER-target gene chromatin. Conversely, DLC1 downregulation compromised the ER transactivation activity and also its nuclear accumulation, suggesting a potential chaperone-like activity of DLC1 in the nuclear translocation of ER. Together, these data define an unexpected upregulation of DLC1 by oestrogen and a previously unrecognized DLC1-ER interaction in supporting and amplifying ER initiated cellular responses in breast cancer cells. PMID- 15891769 TI - Shh directs cell-cycle exit by activating p57Kip2 in the zebrafish retina. AB - The Hedgehog (Hh) family of signalling proteins control both differentiation and proliferation during animal development. Previous studies have shown that Hh signalling has a stimulatory effect on the cell cycle in several organs by controlling core cell-cycle components. Here, we show that Sonic hedgehog (Shh) signalling has the opposite effect in the zebrafish retina, where it leads to cell-cycle exit, and that this is mediated by transcriptional activation of the cyclin kinase inhibitor p57Kip2. The loss of p57Kip2 activity strongly resembles the Shh mutant eye phenotype, and overexpression of p57Kip2 rescues cell-cycle exit in Shh mutants, indicating that p57Kip2 is both necessary and sufficient to mediate Shh-induced cell-cycle exit in the retina. These findings raise the possibility that stimulation of cell-cycle exit through regulation of core cell cycle components may be part of a general mechanism required for Hh-directed differentiation. PMID- 15891770 TI - RNA interference and potential therapeutic applications of short interfering RNAs. AB - RNA interference is an endogenous gene-silencing mechanism that involves double stranded RNA-mediated sequence-specific mRNA degradation. The discovery of this pathway together with the elucidation of the structure and function of short interfering RNAs--the effector molecules of RNA interference--has had an enormous impact on experimental biology. RNA interference technologies are currently the most widely utilized techniques in functional genomic studies. Furthermore, there is an intense research effort aimed at developing short interfering RNAs for therapeutic purposes. A number of proof-of-principle experiments have demonstrated the clinical potential of appropriately designed short interfering RNAs in various diseases including viral infections, cancer and neurodegenerative disorders. Already, in such a short time from their discovery, Acuity Pharmaceuticals (August 2004) and Sirna Therapeutics (September 2004) have filed Investigational New Drug applications with the US FDA to begin clinical trials with modified siRNA molecules in patients with age-related macular degeneration. This review will give a brief overview of the mechanism of RNA interference and applications of the pathway in experimental biology will be discussed. The article will focus on recent developments related to the use of RNA interference technologies in mammalian systems and on potential clinical applications of short interfering RNA-mediated RNA interference. PMID- 15891771 TI - Melanoma-specific expression in first-generation adenoviral vectors in vitro and in vivo -- use of the human tyrosinase promoter with human enhancers. AB - Current treatment regimens for patients with metastatic melanoma are not curative, and new treatment strategies are needed. One possible approach is targeted treatment using the tyrosinase promoter for melanoma-specific expression of genes delivered by adenoviral (Ad) vectors. In this study, a vector with the human minimal tyrosinase promoter and two human enhancer elements (2hE-hTyrP) was compared with different hybrid promoter constructs, containing tyrosinase regulatory sequences and the viral simian virus 40 (SV40) promoter. The tissue specificity of the first-generation vectors was measured by enhanced green fluorescence protein (EGFP) reporter flow cytometry in 12 human melanoma and nonmelanoma cell lines. In the melanotic melanoma cells, the activity of the 2hE hTyrP promoter was comparable with the activity of the cytomegalovirus promoter, and the background expression levels obtained in the nonmelanoma cell lines confirmed the strict tissue-specific property of this promoter. The hybrid SV40 based promoters were effective, but no tissue specificity was observed even after the inclusion of tyrosinase enhancer elements identical to the elements used in the 2hE-hTyrP promoter. The in vivo tissue specificity of the 2hE-hTyrP vector was demonstrated in subcutaneous xenografted tumors by ex vivo detection of EGFP fluorescence with the IVIS Imaging equipment and fluorescence microscopy visualizing the in situ EGFP expression in tumor sections. The tyrosinase mRNA level in the 12 cell lines was measured by quantitative real-time RT-PCR, and the expression levels reliably reflected to what extent the 2hE-hTyrP promoter could drive the gene expression in the individual cell lines. In conclusion, the human tyrosinase promoter fused to two human tyrosinase enhancers (2hE-hTyrP) can be used for efficient tissue-specific expression from first-generation Ad vectors in melanoma cell lines both in vitro and in vivo, as predicted by the quantitative tyrosinase mRNA levels in the melanoma and nonmelanoma cell lines tested. PMID- 15891772 TI - Combined HSV-TK/IL-2 gene therapy in patients with recurrent glioblastoma multiforme: biological and clinical results. AB - Following our pilot clinical study of combined IL-2/HSV-TK gene therapy for recurrent glioblastoma multiforme (GBM), we extended the protocol to a larger population of patients and evaluated safety, feasibility, and biological activity of treatment. A total of 12 patients received intratumor injection of retroviral vector-producing cells (RVPCs), followed by intravenous ganciclovir (GCV). Treatment was well tolerated with only minor adverse events. Transduction of tumor cells was demonstrated in tumor biopsies. A marked and persistent increase of intratumor and plasma Th1 cytokine levels was demonstrated after RVPC injection. At magnetic resonance imaging evaluation, two patients had a partial response (including a patient showing disappearance of a distant noninjected tumor mass), four had a minor response, four had stable disease, and two had progressive disease. The 6- and 12-month progression-free survival rates were 47 and 14%, respectively. The 6- and 12-month overall survival rates were 58 and 25%, respectively. In conclusion, the results of our clinical protocol of gene therapy for recurrent GBM, based on combined delivery of a suicide and a cytokine gene, demonstrate that intratumor injection of RVPCs was safe, provided effective transduction of the therapeutic genes to target tumor cells, and activated a systemic cytokine cascade, with tumor responses in 50% of cases. PMID- 15891773 TI - Gene therapy for hepatocellular carcinoma using sonoporation enhanced by contrast agents. AB - We examined whether sonoporation enhanced by a contrast agent (BR14) was effective in gene therapy for hepatocelluar carcinoma (HCC). Human hepatic cancer cells (SK-Hep1) and plasmid cDNAs expressing green fluorescent protein (GFP), interferonbeta (IFNbeta), and LacZ were used. In vitro, SK-Hep1 cell suspensions with DNA and BR14 were sonoporated. Expressions of every plasmid cDNA and the antitumor effect of IFNbeta were analyzed. In vivo, GFP and IFNbeta genes with BR14 were directly injected into subcutaneous tumors using SK-Hep1 in nude mice, and transcutaneous sonoporation of the tumors was performed. GFP gene transfections and tumor diameters after IFNbeta gene transfection were examined. In vitro, no SK-Hep1 cells were transfected without sonication, whereas transfections were successful after sonication with BR14. Antitumor effect of IFNbeta gene transfection by ultrasound (US) and with BR14 was revealed. In vivo, the SK-Hep1 cells expressed GFP, and the IFNbeta gene transfection by US with BR14 reduced tumor size significantly. In conclusion, gene therapy with sonoporation enhanced by a contrast agent may become a new treatment option for HCC. PMID- 15891774 TI - Development of a gene therapy based bone marrow purging system for leukemias. AB - Although viable gene therapy based methods have been reported for the selective removal or purging of contaminating epithelial derived cancer cells from stem cell grafts, similar strategies for the purging of leukemia cells have been significantly less efficient. Hematopoietic cells are difficult targets for transduction with currently available vectors. Polylysine based molecular conjugate vectors (MCV) were previously found to effectively transduce both normal and malignant hematopoietic cells. A panel of human leukemia cell lines as well as CD34+ selected primary human hematopoietic progenitor cells (HPC) were tested for differential gene expression utilizing different promoters. Reporter gene expression under the control of RSV and SV40 promoters showed a 6-log fold increase in leukemia cells when compared to primary HPC. Using a polylysine based recombinant molecular conglomerate vector (recMCV) encoding the HSV-tk suicide gene under control of RSV, we demonstrated effective and specific cell killing in all leukemia cell lines as well as in primary human leukemia cells derived from chemotherapy refractory patients, while HPC survived under the same conditions at approximately 20% viability. These proof of principle experiments demonstrate that gene therapy technology could be utilized to successfully purge leukemia cells from HPC. PMID- 15891775 TI - Immunity to squamous carcinoma in mice immunized with dendritic cells transfected with genomic DNA from squamous carcinoma cells. AB - Immunotherapy of squamous cell carcinoma (SCC) at an early stage of the disease increases the likelihood of success. We report a new vaccination strategy designed to prepare SCC vaccines from microgram amounts of tumor tissue, enabling the treatment of patients with minimal residual disease. The vaccine was prepared by transfer of sheared genomic DNA-fragments (25 kb) from KLN205 cells, an SCC cell line of DBA/2 mouse origin, into syngeneic bone marrow-derived mature dendritic cells (DCs). More than 90% of the transfected DCs took up DNA from the neoplasm and transferred genes were expressed as protein. The DCs expressed CD11c, CD11b, and the costimulatory molecules CD40, CD80 and CD86, characteristic of mature DCs. Syngeneic DBA/2J mice, highly susceptible to the growth of KLN205 cells, were injected intravenously (i.v.) with the transfected DCs, followed by a subcutaneous (s.c.) injection of the tumor cells. The strong immunogenic properties of the transfected cells were indicated by the finding that the survival of the tumor-bearing mice was prolonged (P<.001), relative to that of mice in various control groups. Enzyme-linked immuno spot (ELISPOT IFN-gamma) assays revealed the activation of cell-mediated immunity directed toward the SCC in mice immunized with the transfected DCs. Two independent in vitro cytotoxicity assays indicated the presence of robust cell-mediated immunity directed toward the SCC in mice immunized with the transfected cells. PMID- 15891776 TI - Folate-linked nanoparticle-mediated suicide gene therapy in human prostate cancer and nasopharyngeal cancer with herpes simplex virus thymidine kinase. AB - For targeted gene delivery to human prostate cancer LNCaP and PC-3 cells and nasopharyngeal cancer KB cells, we developed a folate-linked nanoparticle (NP-F), and evaluated the potential of NP-F-mediated suicide gene therapy in the cells and xenografts with herpes simplex virus thymidine kinase (HSV-tk) and connexin 43 (Cx43). An NP-F-plasmid DNA complex (NP-F nanoplex) showed high DNA transfection efficiency in KB, LNCaP and PC-3 cells. Cell growth inhibition in the presence of ganciclovir (GCV) was enhanced with HSV-tk and Cx43 genes in LNCaP cells. In suicide gene therapy, the tumor growths of KB and LNCaP xenografts were significantly inhibited when an NP-F nanoplex of the HSV-tk gene, and HSV-tk and Cx43 genes, respectively, was injected intratumorally and GCV was administered intraperitoneally. These findings suggested that the NP-F is a potential target vector in prostate and nasopharyngeal cancer for suicide gene therapy. PMID- 15891777 TI - Stress and the brain: from adaptation to disease. AB - In response to stress, the brain activates several neuropeptide-secreting systems. This eventually leads to the release of adrenal corticosteroid hormones, which subsequently feed back on the brain and bind to two types of nuclear receptor that act as transcriptional regulators. By targeting many genes, corticosteroids function in a binary fashion, and serve as a master switch in the control of neuronal and network responses that underlie behavioural adaptation. In genetically predisposed individuals, an imbalance in this binary control mechanism can introduce a bias towards stress-related brain disease after adverse experiences. New candidate susceptibility genes that serve as markers for the prediction of vulnerable phenotypes are now being identified. PMID- 15891778 TI - The neurobiology of zinc in health and disease. AB - The use of zinc in medicinal skin cream was mentioned in Egyptian papyri from 2000 BC (for example, the Smith Papyrus), and zinc has apparently been used fairly steadily throughout Roman and modern times (for example, as the American lotion named for its zinc ore, 'Calamine'). It is, therefore, somewhat ironic that zinc is a relatively late addition to the pantheon of signal ions in biology and medicine. However, the number of biological functions, health implications and pharmacological targets that are emerging for zinc indicate that it might turn out to be 'the calcium of the twenty-first century'. PMID- 15891779 TI - NTP toxicology and carcinogenesis studies of decalin (CAS No. 91-17-8) in F344/N rats and B6C3F(1) mice and a toxicology study of decalin in male NBR rats (inhalation studies). AB - Decalin is used as an industrial solvent for naphthalene, fats, resins, oils, and waxes. It is also used as a substitute for turpentine in lacquers, paints, and varnishes; as a solvent and stabilizer for shoe polishes and floor waxes; and as a constituent of motor fuels and lubricants. Other applications include use as a paint thinner and remover, a patent fuel in stoves, a high-density fuel in submarine-launched cruise missile systems, and in stain removal and cleaning machinery. Decalin was nominated for study by the National Cancer Institute because of its chemical structure, its potential for consumer exposure, and a lack of adequate testing of the chemical. Male and female F344/N rats and B6C3F(1) mice were exposed to decalin (greater than 99% pure) by inhalation for 2 weeks, 3 months, or 2 years. Groups of male NBR rats were exposed to decalin for 2 weeks. Male NBR rats do not produce alpha2u-globulin; the NBR rats were included to study the relationship of alpha2u-globulin and renal lesion induction. Genetic toxicology studies were conducted in Salmonella typhimurium and mouse peripheral blood erythrocytes. 2-WEEK STUDIES IN RATS: Groups of five male and five female F344/N rats and five male NBR rats were exposed to 0, 25, 50, 100, 200, or 400 ppm decalin vapor 6 hours per day, 5 days per week for 16 days. All rats survived to the end of the study, and mean body weights of exposed groups were similar to those of the chamber controls. Renal toxicity studies were performed in male F344/N and NBR rats. The numbers of labeled cells and the labeling indices in the left kidney of 200 and 400 ppm F344/N male rats were significantly greater than those in the chamber controls. The alpha2u globulin/soluble protein ratios were significantly increased in all exposed groups of F344/N rats. Liver weights of male F344/N and NBR rats exposed to 100 ppm or greater were significantly increased, as were those of all exposed groups of females. Kidney weights of male F344/N rats exposed to 50 ppm or greater were significantly increased. Exposure-related hyaline droplet accumulation, degeneration and regeneration of renal cortical tubules, and granular casts occurred in the kidney of exposed F344/N male rats. 2-WEEK STUDIES IN MICE: Groups of five male and five female B6C3F(1) mice were exposed to 0, 25, 50, 100, 200, or 400 ppm decalin vapor 6 hours per day, 5 days per week for 17 days. All mice survived to the end of the study, and mean body weights of exposed groups were similar to those of the chamber control groups. Liver weights of 200 and 400 ppm males and females and 100 ppm females were significantly increased. 3-MONTH STUDY IN RATS: Groups of 25 male and 20 female F344/N rats were exposed to 0, 25, 50, 100, 200, or 400 ppm decalin vapor 6 hours per day, 5 days per week for 2 (five male renal toxicity rats), 6 (10 male and 10 female clinical pathology rats), or 14 (10 core study rats) weeks. All rats survived to the end of the study, and mean body weights of exposed groups were similar to those of the chamber control groups. Urinalysis results indicated that decalin exposure caused increases in urine glucose and protein concentrations and enzyme activities that were consistent with the renal lesions observed microscopically. Renal toxicity studies were performed on rats sacrificed at 2 and 6 weeks and at the end of the study. In kidney tissue examined for cell proliferation, the numbers of PCNA labeled cells and labeling indices were generally significantly greater than those of the chamber controls in exposed groups of rats at all three time points. Concentrations of alpha2u-globulin in the kidney as well as the alpha2u globulin/soluble protein ratios were significantly increased at week 2 in all exposed groups and in the 200 and 400 ppm groups at week 6 and at the end of the study. Absolute and/or relative kidney and liver weights of male rats exposed to 50 ppm or greater were increased. Incidences of renal tubule regeneration and granular casts in the medulla of the kidney in exposed male rats were increased, and the severities of hyaline droplets generally increased with increasing exposure concentration. 3-MONTH STUDY IN MICE: Groups of 10 male and 10 female B6C3F(1) mice were exposed to 0, 25, 50, 100, 200, or 400 ppm decalin vapor 6 hours per day, 5 days per week for 14 weeks. All mice survived to the end of the study, and mean body weights of exposed groups were similar to those of the chamber control groups. Liver weights of 200 and 400 ppm males and females were significantly increased. There was a significant exposure concentration-related decrease in the absolute spermatid head count and a significant decrease in absolute head count of the 400 ppm group compared to the chamber controls. Incidences of centrilobular cytomegaly of the liver were increased in exposed male mice. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female F344/N rats were exposed to 0, 25, 50 (male rats only), 100, or 400 ppm (female rats only) decalin vapor 6 hours per day, 5 days per week for 105 weeks. A group of 20 male rats was exposed to 400 ppm. Survival of exposed groups was similar to that of the chamber control groups. Mean body weights of 400 ppm males were slightly less than those of the chamber controls during the second year of the study. Incidences of renal tubule adenoma and adenoma or carcinoma (combined) and of benign or malignant pheochromocytoma (combined) of the adrenal medulla in 100 and 400 ppm males were significantly increased. There was a significant association between nephropathy severity and adrenal pheochromocytoma incidence. Nonneoplastic lesions related to decalin exposure occurred in the kidney of male rats. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female B6C3F(1) mice were exposed to 0, 25, 100, or 400 ppm decalin vapor 6 hours per day, 5 days per week for 105 weeks. Survival of exposed mice was similar to that of the chamber controls. Mean body weights of exposed groups were generally similar to those of the chamber control groups throughout the study. Increased incidences of hepatocellular neoplasms occurred in 25 and 400 ppm female mice, and the incidences of centrilobular hypertrophy, necrosis, syncytial alteration, and erythrophagocytosis of the liver in 400 ppm males were significantly increased. The incidences of uterine stromal polyp and stromal polyp or stromal sarcoma (combined) occurred with positive trends in female mice. PHARMACOKINETIC MODEL: The rate of metabolism of decalin was the same for males and females in rats and mice. Also in rats and mice, decalin metabolism was saturated at less than 400 ppm. Increased labeling indices in male rats were likely due to changes related to alpha2u-globulin. GENETIC TOXICOLOGY: Decalin was not mutagenic in S. typhimurium strains TA97, TA98, TA100, or TA1535, with or without induced hamster or rat liver S9 enzymes. A small but significant increase in the frequency of micronucleated normochromatic erythrocytes was noted in male mice exposed to decalin for 3 months; however, no induction of micronuclei was observed in female mice. CONCLUSIONS: Under the conditions of these studies, there was clear evidence of carcinogenic activity of decalin in male F344/N rats based on increased incidences of renal tubule neoplasms. The increased incidences of benign or malignant pheochromocytoma (combined) of the adrenal medulla in male rats were also considered to be exposure related. There was no evidence of carcinogenic activity of decalin in female F344/N rats exposed to 25, 100, or 400 ppm. There was no evidence of carcinogenic activity of decalin in male B6C3F(1) mice exposed to 25, 100, or 400 ppm. There was equivocal evidence of carcinogenic activity of decalin in female B6C3F(1) mice based on marginally increased incidences of hepatocellular and uterine neoplasms. Exposure of male rats to decalin resulted in nonneoplastic lesions of the kidney characteristic of alpha2u-globulin accumulation. Nonneoplastic lesions of the liver were observed in male mice exposed to decalin. PMID- 15891780 TI - Toxicology and carcinogenesis studies of malachite green chloride and leucomalachite green. (CAS NOS. 569-64-2 and 129-73-7) in F344/N rats and B6C3F1 mice (feed studies). AB - BACKGROUND: Malachite green chloride is a dye used to prevent fungus infections in commercial fisheries. Leucomalachite green is formed from malachite green and remains in the tissues of exposed fish. We studied the effects of malachite green on female rats and female mice, and the effects of leucomalachite green on male and female rats and female mice, to identify potential toxic or cancer-related hazards to humans. METHODS: For each study we mixed the dye into the feed of rats and mice. The doses of malachite green chloride given were 100, 300, or 600 parts per million (ppm) for female rats and 100, 225, or 450 ppm for female mice. Doses of leucomalachite green were 91, 272, or 543 ppm for male and female rats and 91, 204, or 408 ppm for female mice. There were 48 animals in each dose group. Control animals received the same feed with no chemical added. The study lasted for two years. Tissues from more than 40 sites were examined for every animal. RESULTS: Rats, but not mice, exposed to malachite green chloride or leucomalachite green weighed less on average than the control animals. In rats exposed to the dyes, there were very slight increases in a few types of tumors: cancers of the thyroid gland, liver, and mammary gland in females exposed to malachite green chloride; of the thyroid gland and testes in males exposed to leucomalachite green; and of the thyroid gland and liver of females exposed to leucomalachite green. We saw no increase in cancers in female mice given malachite green chloride, but there was an increase in liver tumors in female mice given to leucomalachite green. CONCLUSIONS: We conclude that tumors of the thyroid gland, liver, or mammary gland in female rats might have been caused by malachite green chloride, but the malachite green chloride did not cause cancer in female mice. We conclude that leucomalachite green might have caused cancers of the thyroid gland in male and female rats, and of the testes in male rats and liver in female rats. Leucomalachite green caused an increase in cancer of the liver in female mice. PMID- 15891782 TI - Orthodontic specialists education in Europe: past, present and future. AB - The field of orthodontics developed simultaneously in the USA and Europe, but along somewhat different routes. It is explained that structural changes in Europe resulting in an EU with 25 countries have led to the freedom of exchange of professionals among the Member States which were participating in the EU before May 1, 2004. The European harmonization of dental education is explained. Subsequently, the training of orthodontic specialists in the USA and Europe is dealt with. Furthermore an overview of the Erasmus Programme is given. The role and structure of the European Federation of Orthodontic Specialists Associations (EFOSA) is clarified. The current state of orthodontic specialty training in Europe is described in detail with emphasis on training aspects. Finally, some suggestions are made for the improvement of the training of orthodontic specialists. PMID- 15891783 TI - If wishes were horses: functional appliances and growth modification. AB - OBJECTIVES: Aside from the common observation that functional appliances work, the literature has little to say about the means by which a correction is achieved and whether or not it differs from that produced by a single phase of fixed-appliance treatment. There seems to be relatively little interest in these basic questions. The purpose of this communication is to explore and discuss the action of functional appliances in light of their interaction with the gross mechanisms of craniofacial growth. It is hoped that a collegial discussion will ensue. RESULTS: Normally, the mandible outgrows the midface, even in Class II patients. This favorable pattern of growth, however, produces maxillary dentoalveolar compensations that prevent the excess mandibular growth from having any marked effect on the malocclusion. Functional appliances may avoid this mesial movement of the maxillary dentition by dissociating mandibular displacement from the condylar growth that normally accompanies it. By producing an anterior functional shift that corrects the Class II relationships all at once and by maintaining this advanced position long enough for the condyles to grow back into the fossae, functional appliances can make clever use of the normal pattern. No "extra" growth may be necessary. Relative to untreated controls, their ultimate effect is in the midface, rather than the mandible. Moreover, the final correction may not differ significantly from that produced by fixed appliances or by two-phase treatments in which the first phase is some sort of extra-oral traction to the midface. CONCLUSION: Functional appliances may utilize the normal pattern of facial growth, rather than some sort of mandibular "growth modification," to achieve a molar correction. As a result, their ultimate effect may not differ from that produced by the various one- and two-stage alternatives. The choice of which to use may well prove to be a simple practice-management decision. PMID- 15891784 TI - Obstructive Sleep Apnoea Syndrome (OSAS) and rhino-tubaric disfunction in children: therapeutic effects of RME therapy. AB - The aim of the study was to evaluate if RME (Rapid Maxillary Expansion) therapy could improve both the patency of the nasal airways and the Obstructive Sleep Apnoea Syndrome (OSAS). 42 children with a case history of oral breathing, snoring and night time apnoeas were studied. Selection criteria were: no adenotonsillar hypertrophy, Body Mass Index (BMI) below 24 and a malocclusion characterised by a narrow upper jaw, determined by postero-anterior cephalometric evaluation. Patients underwent an ENT visit with auditory and respiratory tests including daytime a sleepiness questionnaire, a 19-channel polysomnography, and an orthognatodontic examination; finally the patient underwent X-rays investigations. All the investigations were carried out before orthodontic therapy (T0), after one month (T1) with the device still on, and 4 months after the end of the orthodontic treatment which lasted for about 6-12 months (T2). All the changes induced by RME on the upper jaw and nasal septum were analysed by postero-anterior cephalometric evaluation in T0, T1 and T2. In all treated cases, the authors obtained an opening of the midpalatal suture; this was confirmed both by intraoral occlusal X-rays and postero-anterior cephalograms. The results reported by the 42 patients studied show that the R.M.E. therapy widens the nasal fossa and releases the septum thus restoring a normal nasal airflow with disappearance of obstructive sleep disordered breathing. Changing the anatomic structure, RME brought a significant functional improvement. Therefore the orthodontist can play an important role in the interdisciplinary treatment of OSAS patients. PMID- 15891785 TI - Implants in orthodontics. Interview. PMID- 15891786 TI - Spider Screw: skeletal anchorage system. AB - The stability of the anchorage unit plays a very important role in orthodontic control. Controlled orthodontic movements such as retraction and/or protraction of teeth and intrusion of overerupted teeth are very difficult to achieve without patient cooperation and without causing undesirable reciprocal movement in the anchorage unit. The article describes characteristics, surgical procedure, and clinical use of the Spider Screw as an ideal non-dental and non-cooperation based anchorage system. The Spider Screws are self-tapping, titanium mini-screws with immediate loading capability. Their utilization involves a simple biomechanical principle combined with the utilization of minimum orthodontic mechanotherapy. Ideal orthodontic forces (in the range from 50 to 250 gr) can be applied to achieve the desired orthodontic movements. Complete osteointegration is neither expected nor desired with this anchorage system. The Spider Screw anchorage system can be used to support a variety of orthodontic movements specifically in clinical situations involving incomplete dental arches and limited cooperation as in many adult orthodontic cases. The ease of surgical placement combined with the reduced dimension of the Spider Screw diameter equally permits its use in clinical situations where anchorage recovery is necessary during treatment of complete dentitions in classical orthodontic therapy. PMID- 15891787 TI - Mechanical properties of three different commercially available miniscrews for skeletal anchorage. AB - BACKGROUND: During the last 5 years, anchorage control with self-tapping miniscrews has become an important part of the clinical management of orthodontic patients. Yet, no studies have been performed for measuring mechanical properties of the currently available systems. OBJECTIVES: The purpose of this study is the evaluation of mechanical properties of three commercially available self-tapping screw systems used in orthodontic treatment. MATERIALS: three systems with a 1.5 mm diameter and 11 mm length screw (Leone, Firenze, Italy; M.A.S. Micerium, Avegno, Italy; Dentos, Korea) were examined. The results compared the resistance to bending, torque, pullout of each screw and the insertion moments needed to screw down each sample. CONCLUSIONS: All three miniscrews have mechanical properties that contribute to their safe use as skeletal anchorage in orthodontics. Although stainless steel has demonstrated to be more resistant to failure than titanium, its overall performance as material for miniscrews could be inferior to titanium. In order to facilitate the insertion, the asymmetric profile of the thread should be preferred to the symmetric cut. The ratio between the diameter of the drill and the diameter of the corresponding miniscrew is pivotal for the successful implantation and resistance of the miniscrews. Looking at the mechanical properties evaluated in this study, a cylindric shape of the screw is better than a conic one. The conic shape could be preferred in case the site of insertion is iterradicular and therefore limited to 2.5-3.5 millimetres. PMID- 15891789 TI - Italian Board of Orthodontics: case N. 2 adult malocclusion. AB - OBJECTIVES: this 20,2 year-old girl presented with a class III malocclusion with severe crowding. MATERIALS AND METHODS: she was at the end of her growth with a severe skeletal and dental class III malocclusion with lower midline deviation and severe crowding. A concave profile, due to chin's prominence, was present. Lower vertical third of the face increased. No signs or symptoms of TMJ problems were present: lingual position of 12 and 22 is a potential problem for TMJ's health. The state of oral mucosa and gingiva was good. But oral hygiene was not good. Lower midline deviated 3 mm to the left side; canine and molar class I on the right side and class III on the left side. The sequence of her treatment was as follows: a) extraction of 15, 25, 35, 44; b) upper and lower arch fixed appliance for alignment, leveling, correction of lower midline and occlusal relationship; c) post-treatment retention. PMID- 15891790 TI - Polymer-supported manganese porphyrin catalysts--peptide-linker promoted chemoselectivity. AB - Manganese porphyrin catalysts were tethered to polymer-supports via peptide linkers. The reactivity and chemoselectivity of the catalysts were assessed in the epoxidation of limonene. It was found that the inclusion of a peptide linker incorporating a donor heteroatom which could act as an axial ligand led to a supported manganese porphyrin catalyst with unprecedented selectivity and stability. PMID- 15891791 TI - [Francisco Montiel Avendano (1934-2004): remembrance and homage to a great friend]. PMID- 15891792 TI - The Vibrio parahaemolyticus pandemic. PMID- 15891793 TI - [Review and guidelines for treatment of diarrhea caused by Vibrio parahaemolyticus]. AB - In Chile Vibrio parahaemolyticus has been detected in 3 gastroenteritis outbreaks since 1998. The most recent outbreak occurred during the summer of 2005, affecting over 10,000 people of whom one died. Affected individuals presented with one or more of the following symptoms: diarrhea, nausea, vomiting, abdominal pain and/or fever. Fecal white blood cells were detected in only 6% of patients. The predominant serotype in the 3 outbreaks was the pandemic O3:K6 strain. Diagnosis was confirmed by isolation and identification of V. parahaemolyticus in stool cultures and/or by establishing an epidemiological link. V. parahaemolyticus isolates were 100% susceptible to tetracycline, ciprofloxacin and chloramphenicol, and universally resistant to ampicillin. Due to the public health impact of the 2005 outbreak, the Ministry of Health called for a National Task Force mandated to review epidemiological, clinical and microbiological features of the outbreak and to propose management guidelines. PMID- 15891794 TI - [Search of amantadine-resistance in influenza A strains isolated in Santiago, Chile, 2001-2002]. AB - Amantadine has been used for prevention and treatment of influenza A infection. It blocks the proton through the M2 ion channel. Drug-resistant viruses appear quickly when this therapy is used. Single amino acids changes in the H2 protein can confer resistance, being the most frequent one in position 31. Different methods to detect resistant strains have been described. The objectives were to determine the existence of amantadine resistance of influenza A strains isolated in a virologic laboratory in Santiago, Chile, between 2001-2002, and to validate a new molecular method to detect resistant strains. A PCR restriction fragment length polymorphism analysis was employed for the detection of resistant viruses. In 31 processed strains no mutation in the position 31 was found. This result supports that amantadine resistance is very low or absent in Chile. This could be explained by a limited use of this drug in the study population. This method could be used as a monitoring system to survey resistant viruses. PMID- 15891795 TI - [Sore throat, streptococcal etiology? Use of etiological screening and antimicrobial prescription in two university pediatric centers]. AB - The diagnosis and treatment of sore throat in pediatric patients attending one of two facilities at a university hospital: pediatric ambulatory facility (PAF) and emergency unit (EU) in a period of 14 months are described. There were 421 patients at the PAF and 289 at the EU, of whom 65% and 76% were studied for Streptococcus pyogenes respectively. Streptococcus were detected in both centers in 37% of cases, with a peak detection of 50% at ages 6 to 10 years. There were 217 patients without any bacteriological study, of whom 162 (75%) received antibiotics. The importance of bacteriological confirmation prior to antibiotic prescription in this disease is emphasized. PMID- 15891796 TI - [Necrotizing pneumonia due to Rhodococcus equi in non HIV immunocompromised host. Case report and review]. AB - Rhodococcus equi, is a grampositive intracellular bacillus, that causes infection mainly in immunocompromised hosts. We report the case of a 52 years old woman, with a systemic lupus erythematosus and a progressive 10 months evolution with cough, dyspnea, mucous-purulent sputum, occasionally hemoptysis, intermittent fever, and weight loss of 10%. With partial response to antibiotic treatment, radiological evolution of thoracic scanners evidenced the development of multiple bilateral areas of consolidation, some of them nodular. Percutaneous thoracic biopsy showed characteristic histology and microbiological analysis yielded the identification of Rhodococcus equi in both bronchoalveolar lavage and pulmonary biopsy. She received prolonged bi-associated antibiotic treatment with good clinical and radiological response. This agent must be considered in the study of immunocompromised patients that present with a prolonged evolution of pneumonia. PMID- 15891797 TI - [Urinary tract infection in childhood]. AB - The urinary tract infection (UTI) is one of the major causes of bacterial infection in children. Therefore an opportune diagnosis, an adequate treatment and a close follow up will prevent chronic renal damage. In this brief article UTI clinical characteristics and recommendations for diagnosis and treatment according to children age are reviewed. PMID- 15891798 TI - [Epidemiology of HIV/AIDS in Chile. December 2003]. AB - From 1984, when the first case of AIDS was diagnosed in Chile, to December 2003, 6.060 patients with AIDS and 6.514 with asymptomatic HIV infection have been notified to a passive national surveillance system; around 3,800 have died due to the infection. Magnitude of the under reporting is discussed. Assumed routes of the infection have been: sexual (94.1%), blood--largely intravenous drug use- (4.3%) and vertical transmission (1.6%). According to analysis performed by the National Commission on AIDS (CONASIDA), a governmental office, HIV/AIDS epidemic in Chile is characterized by: predominance in male homo/bisexuals, urban and rural distribution, impoverishment of the affected people, diagnosis made mainly during adulthood and a steady trend to affecting more women and heterosexuals at large. Since 2001 there has been a continuous increase in access to standard antiretroviral therapy (HAART) for those cared for by the public health system, reaching free coverage for 100% of this population by 2003, which has determined a significant decrease in the rate AIDS-associated clinical manifestations while reaching an stop of the previously increasing lethality of the infection (from a 15% increase from 1993-1997 to 0.2% from 1998-2003). PMID- 15891799 TI - [Figures in Chilean infectology. Professor Doctor Roque Kraljevic Orlandini]. PMID- 15891800 TI - [The El Progreso Medico journal and the Berlin meeting in 1890]. PMID- 15891801 TI - [Rapid methods for the diagnosis of influenza]. PMID- 15891803 TI - Multiseptate gallbladder: MR and MR cholangiographic findings. PMID- 15891804 TI - Adenocarcinoma of the ileum in Crohn disease. AB - Small bowel adenocarcinoma develops in 1.5% of patients who have longstanding Crohn disease and is very rarely diagnosed preoperatively because of its rarity, overlapping imaging features, and lack of reported cases. Nonspecific findings including loss of mural stratification (i.e., "target sign") and mild degree of bowel wall enhancement when combined with enlarged mesenteric lymph nodes were helpful computed tomographic findings to suspect malignancy in our case. PMID- 15891805 TI - Dual-phase multidetector thin-section CT in detecting duodenal gastrinoma. AB - Preoperative localization of gastrinomas, especially of extrapancreatic origin, remains a challenge to the radiologist. Most patients with extrapancreatic gastrinomas undergo surgery without preoperative identification of the primary tumor. The appropriate imaging modality to localize gastrinomas is under continuing debate. We report a case of a duodenal gastrinoma with regional lymph node metastases that presented with Zollinger-Ellison syndrome. The small primary tumor was detected noninvasively by dual-phase multidetector thin-section computed tomography with adequate bowel distention and confirmed by endoscopy and histopathologic examination. The case illustrates that appropriate computed tomographic technique and scanning protocol are crucial for success in localizing extrapancreatic gastrinoma. PMID- 15891806 TI - Routine symptom screening for postnatal urinary and anal incontinence in new mothers from a district. AB - Parous middle-aged women present with urinary and faecal incontinence and childbirth trauma is thought to be a causal factor. Both symptoms are common after childbirth but usually go under-reported. It has been suggested that new mothers are likely to benefit from routine symptom screening because by actively identifying symptomatic women they could then be helped to access continence services. The main objective of this study was to pilot a programme of routine symptom screening for postnatal urinary and anal incontinence in new mothers from a district general hospital. Self-completion questionnaires for both urinary and anal incontinence were sent by post to 442 primiparous women who had delivered consecutively 10 months previously in a district general hospital. Two hundred and seventy-five new mothers responded to the survey. Ninety-two women had new symptoms of incontinence at 10 months, 82 had urinary incontinence and 25 had anal incontinence. However, only six new mothers had discussed the problem with a health professional. Although nine women requested a hospital appointment none of the women attended the appointment arranged for them. The domain scores on both questionnaires were significantly less for symptomatic new mothers when compared to women with established symptoms of incontinence. The programme of screening successfully identified women with symptoms of incontinence. However, all of the symptomatic women declined a follow-up appointment at hospital which questions the benefits of routine screening 10 months after childbirth. PMID- 15891807 TI - Isolated glossopharyngeal and vagus nerves palsy due to fracture involving the left jugular foramen. AB - This report describes a case of delayed post-traumatic glossopharyngeal and vagus nerves palsy (i.e. dysphonia and swallowing dysfunction). A high resolution CT study of the cranial base detected a fracture rim encroaching on the left jugular foramen. Treatment consisted in supportive measures with incomplete recovery during a one-year follow-up period. Lower cranial nerves palsies after head trauma are rare and, should they occur, a thorough investigation in search of posterior cranial base and cranio-cervical lesions is warranted. The presumptive mechanism in our case is a fracture-related oedema and ischemic damage to the nerves leading to the delayed occurrence of the palsy. PMID- 15891808 TI - Gamma knife surgery for brain metastases: indications for and limitations of a local treatment protocol. AB - OBJECTIVE: The purpose of this retrospective study was to evaluate results of a local treatment protocol using gamma knife surgery (GKS) for brain metastases without upfront whole brain radiation therapy (WBRT). METHODS: Results for 521 consecutive patients satisfying the following 3 criteria were analysed: 1) a maximum of 3 tumours with a diameter of 25 mm or more; 2) no prior WBRT; 3) no surgically in accessible large (>30 mm) tumours. Large tumours were surgically removed and all smaller lesions were treated by GKS without up front WBRT. New lesions, detected with follow-up MRI, were appropriately treated with repeat GKS. Overall survival (OS), neurological survival (NS), qualitative survival (QS) and new lesion-free survival (NLFS) curves were calculated and the prognostic values of covariates were obtained. OS and NS were compared according to tumour number. RESULTS: In total, 1023 separate sessions were required to treat 4562 lesions. The primary organs were lung in 369 patients, gastro-intestinal tract in 70, breast in 33, urinary tract in 24, and others/unknown in 25. The median OS period was 9.0 months. On multivariate analysis, the significant prognostic factors for OS were found to be extracranial disease (risk factor: active), Karnofsky performance status (KPS) score (<70) and gender (male). NS and QS at one year were 85.6% and 73.0%, respectively. The only significantly poor prognostic factor for NS was carcinomatous meningitis. NLFS at 6 months was 68.9%. For both OS and NS, the differences between a few (10) tumours had a significantly poorer prognosis than those with 0.05). Median follow-up was 4.5 years (range 1-8 years). Recurrence rates for the suture and mesh repair groups were 11.5 and 0%, respectively (P=0.007). Infection rates for the suture and mesh repair groups were 11.5 and 0%, respectively (P=0.007). Our data suggest that prosthetic mesh repair is ideal for managing primary and recurrent umbilical hernias in both obese and non-obese patients. PMID- 15891811 TI - Lichtenstein inguinal hernioplasty: sutures versus glue. AB - INTRODUCTION: This was a prospective, randomised study undertaken from January 2001 to July 2003 firstly to assess the possibility of using glues to fix meshes, secondly to verify whether or not this leads to a higher hernia recurrence rate, and thirdly to assess post-operative pain. METHODS: This study covered a total of 55 patients with bilateral hernias who presented similar characteristics on both sides and on whom fixation of the mesh was undertaken differently, depending on the side. On the right side polypropylene sutures were used (prolene 2/0), while on the left, attachment was done using glue (Tissucol). RESULTS AND CONCLUSIONS: Results were similar in both inguinal regions, although there was less post operative pain and less inflammatory reaction on the left side. There was no hernia recurrence in the follow-up at the end of 1 year. PMID- 15891812 TI - Coskun repair of inguinal hernia: is it really new? PMID- 15891813 TI - Surgeon perspectives on options for ventral abdominal wall hernia repair: results of a postal questionnaire. AB - BACKGROUND: Ventral abdominal wall hernias are a common cause of morbidity and mortality. Opinion varies as to appropriate management. A recent consensus meeting on incisional hernia identified the need to standardise repair. On this background, a survey of current practice was performed. METHOD: A questionnaire was sent to 101 practicing general surgeons within the West of Scotland. Incisional, epigastric and para-umbilical defects were subdivided into defect size <2, 2-5 and >5 cm. The surgeons were asked to indicate the most appropriate repair (suture, mayo or mesh) for each. The influence of reducibility on the decision to repair was also assessed. RESULTS: Sixty-one of 101 questionnaires were returned valid giving a response rate of 60%. Suture repair was significantly more likely to be used in all defects <2 cm (P<0.001). Mesh repair was significantly more likely to be recommended in all defects >5 cm (P<0.001). Of defects >5-cm, mesh was recommended for 90% of incisional hernia compared with 81% of epigastric and 76% of para-umbilical hernia (P<0.001). There was no significant difference in choice of repair for defect size 2-5 cm with opinion divided between suture and mesh. Irreducibility increased the likelihood of recommendation for repair. CONCLUSION: This survey shows a lack of consensus on the appropriate repair of ventral abdominal wall hernia among practicing consultant general surgeons. This reflects the contrasting views within the current literature. PMID- 15891814 TI - Effects of climate variation on timing of nesting, reproductive success, and offspring sex ratios of red-winged blackbirds. AB - Predicting ecological consequences of climate change will be improved by understanding how species are affected by contemporary climate variation, particularly if analyses involve more than single ecological variables and focus on large-scale climate phenomena. I used 18 years of data from red-winged blackbirds (Agelaius phoeniceus) studied over a 25-year period in eastern Ontario to explore chronological and climate-related patterns of reproduction. Although blackbirds started nesting earlier in years with warmer springs, associated with low winter values of the North Atlantic Oscillation Index (NAOI), there was no advance in laying dates over the study. Nesting ended progressively later and the breeding season lasted longer over the study, however, associated with higher spring values of NAOI. As the length of the nesting season increased, offspring sex ratios became more female biased, apparently as a result of females adjusting the sex of the eggs they laid, rather than from sex-biased nestling mortality. Clutch size did not vary systematically over the study or with climate. Opposing trends of declining nest success and increasing productivity of successful nests over the study resulted in no chronological change in productivity per female. Higher productivity of successful nests was associated with higher winter NAOI values, possibly because synchrony between nesting and food availability was higher in years with high NAOI values. Other than the association between the start of nesting and spring temperatures, local weather (e.g., temperature, rainfall) patterns that linked NAOI with reproduction were not identified, suggesting that weather patterns may be complex. Because climate affected most aspects of red-winged blackbird reproduction examined, focusing on associations between climate and single variables (e.g., first-egg dates) will have limited value in predicting how future climates will affect populations. PMID- 15891815 TI - Extreme isotopic depletion of nitrogen in New Zealand lithophytes and epiphytes; the result of diffusive uptake of atmospheric ammonia? AB - Several lichens and the terrestrial alga Trentepohlia were found to have extremely depleted 15N signatures at two sites near the Rotorua geothermal area, New Zealand. Values, typically -20 per thousand, with several extreme cases of 24 per thousand, are more isotopically depleted than any previously quoted delta15N signature for vegetation growing in natural environments. For Trentepohlia, distance from a geothermal source did not affect isotopic signature. A 100-km transect showed that the phenomenon is widespread and the discrimination is not related to substrate N, or to elevation. Rainfall NHx and atmospheric gaseous NH3 (NH3(g)) were shown to be isotopically depleted in the range -1 per thousand to -8 per thousand and could not, of themselves, be responsible for the plant values obtained. A simulation of Trentepohlia thallus was created using an acidified fiberglass mat and was allowed to absorb NH3(g) from the atmosphere. Mats exposed at the geothermal sites and on farm-land showed a significant further depletion of 15N to -17 per thousand. We hypothesize that the extreme isotopic depletion is due to dual fractionation: firstly by the volatilization of NH3(g) from aqueous sources into the atmosphere; secondly by the diffusive assimilation of that NH3(g) into vegetation. We further hypothesize that lithophytes, epiphytes, and higher plants, growing on strongly N-limited substrates, will show this phenomenon more or less, depending on the proportion of diffusively assimilated NH3(g) utilized as a N source. Many of the isotopically depleted delta15N signatures in vegetation, previously reported in the literature, especially epiphytes, may be due to this form of uptake depending on the concentration of atmospheric NH3(g), and the degree of reliance on that form of N. PMID- 15891816 TI - Plant population size and isolation affect herbivory of Silene latifolia by the specialist herbivore Hadena bicruris and parasitism of the herbivore by parasitoids. AB - Habitat fragmentation can affect levels of herbivory in plant populations if plants and herbivores are differentially affected by fragmentation. Moreover, if herbivores are top-down controlled by predators or parasitoids, herbivory may also be affected by differential effects of fragmentation on herbivores and their natural enemies. We used natural Silene latifolia populations to examine the effects of plant population size and isolation on the level of herbivory by the seed predating noctuid Hadena bicruris and the rate of parasitism of the herbivore by its parasitoids. In addition, we examined oviposition rate, herbivory and parasitism in differently sized experimental populations. In natural populations, the level of herbivory increased and the rate of parasitism decreased with decreasing plant population size and increasing degree of isolation. The number of parasitoid species also declined with decreasing plant population size. In the experimental populations, the level of herbivory was also higher in smaller populations, in accordance with higher oviposition rates, but was not accompanied by lower rates of parasitism. Similarly, oviposition rate and herbivory, but not parasitism rate, increased near the edges of populations. These results suggests that in this system with the well dispersing herbivore H. bicruris, habitat fragmentation increases herbivory of the plant through a behavioural response of the moth that leads to higher oviposition rates in fragmented populations with a reduced population size, increased isolation and higher edge-to-interior ratio. Although the rate of parasitism and the number of parasitoid species declined with decreasing population size in the natural populations, we argue that in this system it is unlikely that this decline made a major contribution to increased herbivory. PMID- 15891817 TI - Within-plant variation in induced defence in developing leaves of cotton plants. AB - According to optimal defence theory (ODT) plants should invest in stronger defence in the most valuable parts, such as reproductive or young tissue. Cotton plants are known to express high resistance to herbivore feeding in the developing leaves at the top of the plant after herbivore-damage. Cotton plants also have developing leaves on side shoots from the nodes all along the plant. This allowed us to investigate within-plant distribution of defence between younger tissues at different locations on the plant. We found that all developing leaves showed increased resistance to feeding by larvae of the generalist moth Spodoptera littoralis after herbivore damage to leaves of the plant. An increase in the concentration of secondary compounds, terpenoid aldehydes, was found in developing leaves both at the top of the plant and on the side shoots. However, the resistance was stronger in the top leaves than in the side shoot leaves, showing that there is intra-plant variation in the induced response between different leaves of the same age. After the initial damage, larval feeding mainly occurred on the older, fully developed true leaves. Furthermore, the herbivore induced resistance in the developing leaves reduced upward movement of feeding larvae on the plant and reduced the feeding on the upper parts of the plant over a period of at least 6 days. The plant thus benefits from defending all developing leaves by directing feeding to older, less valuable leaves and lower parts of the plant. The observed distribution of defence within cotton plants supports ODT. PMID- 15891818 TI - Dose and host characteristics influence virulence of ranavirus infections. AB - Parasites play a prominent role in the ecology, evolution, and more recently, conservation of many organisms. For example, emerging infectious diseases, including a group of lethal ranaviruses, are associated with the declines and extinctions of amphibians around the world. An increasingly important basic and applied question is: what controls parasite virulence? We used a dose-response experiment with three laboratory-bred clutches of tiger salamander larvae (Ambystoma tigrinum) to test how the size of inoculum and host genetic factors influence the dynamics and outcome of ranavirus infections. We found that infection rates increased with dose and were strongly affected by clutch identity and host life history stage. Case mortality increased with dose of inoculum, but was unaffected by host characteristics. Average survival time decreased with dose and differed among clutches, but this was largely due to differences in the time to onset of symptoms. Overall, our results suggest that dose of inoculum and host characteristics (life history stage and genetic background) influence the establishment and early virus replication, and therefore the virulence of ranavirus infections. PMID- 15891819 TI - What determines interspecific variation in relative growth rate of Eucalyptus seedlings? AB - The present study examines relative growth rate (RGR) and its determinants in seedlings of nine Eucalyptus species. Species were selected from mesic (1,800 mm a(-1) rainfall) through to semi-arid habitats (300 mm a(-1)), and thus, notionally vary in "stress" tolerance. Seedlings were grown in a glasshouse during early summer and received between 33 mol and 41 mol PAR m(-2) day(-1) . The mean RGR varied among species-from a minimum of 66 mg g(-1) day(-1) in E. hypochlamydea to a maximum of 106 mg g(-1) day(-1) in E. delegatensis. RGR was positively related to rainfall at the sites of seed collection. Neither specific leaf area (SLA) nor net assimilation rate was related to rainfall or RGR. While the absence of relationships with SLA and net assimilation rate contrasts with other studies and species, we cannot rule out the effects of sample size (n=9 species) and modest ranges in SLA and RGR. The ratio of leaf mass to total mass (LMR) varied from 0.49+/-0.07 g g(-1) in E. socialis to 0.74+/-0.04 g g(-1) in E. delegatensis and was strongly positively related with rainfall (r2=0.77). Interspecific differences in RGR were strongly related to LMR (positive relationship, r2=0.50) and the rate of dry matter production per mol of leaf nitrogen (positive relationship, r2=0.64). Hence, the slow RGR of low-rainfall species was functionally related to a lower growth rate per mol of leaf nitrogen than high-rainfall species. Furthermore, slow RGR of low-rainfall species was related to greater allocation to roots at the expense of leaves. Increasing allocation to roots versus leaves is likely an adaptation to soil and atmospheric water deficits, but one that comes at the expense of a slow RGR. PMID- 15891820 TI - Life history consequences of sterol availability in the aquatic keystone species Daphnia. AB - The absence of essential biochemical nutrients, such as polyunsaturated fatty acids or sterols, has been considered as a mechanism determining trophic interactions between the herbivore Daphnia and its phytoplankton food source. Here, we experimentally quantify the sensitivity of two Daphnia species to decreasing amounts of dietary sterols by measuring variations in life history traits. The two species Daphnia magna and D. galeata were fed different mixtures of the sterol-containing green alga Scenedesmus obliquus and the sterol-free cyanobacterium Synechococcus elongatus; a higher proportion of Synechococcus in the food is equivalent to a decrease in dietary sterols. To address the significance of sterol limitation, the Daphnia species were also fed Synechococcus supplemented with cholesterol. In both species, somatic and population growth rates, maternal dry mass, the number of viable offspring, and the probability of survival were significantly reduced with the lower availability of sterols. A high correlation between the sterol content of the mixed diet and the somatic and population growth rates was found, and growth on cholesterol-supplemented Synechococcus fitted well into this correlation. Somatic growth of first-clutch neonates grown on 100% Synechococcus exhibited a pattern similar to that of somatic growth of their mothers grown on the different food regimes, which demonstrated the significance of maternal effects for sterol limited population growth. Daphnia galeata had a twofold higher incipient limiting sterol level than D. magna, which indicated interspecific differences in sterol requirements between the two Daphnia species. The results suggest a strong impact of dietary sterols on life history traits and therefore, population dynamics of the keystone species Daphnia. PMID- 15891821 TI - Experimental removal of the male parent negatively affects growth and immunocompetence in nestling great tits. AB - Given the available empirical evidence on the benefits and costs associated with immune defence, a role for the immune system in the trade-off between current and future reproduction has been predicted. This hypothesis was studied in a free living population of great tits (Parus major) by examining the effects of male removal on the immunocompetence, body condition, and recapture probability in the widowed females and their nestlings. Furthermore, we investigated whether growth and its relation to immunocompetence were affected in the nestlings. For a short lived species such as the great tit, one could predict that widowed females will compensate for the lack of any male assistance in feeding of their chicks and that they consequently might jeopardize their own health. However, we did not find any negative effects of male removal on body mass or condition, nor on humoral immunocompetence against sheep red blood cells in the widowed females by the end of the feeding period. In contrast, we observed significantly reduced body mass and size as well as a reduced T-lymphocyte cell-mediated immune response (expressed as the thickness of the swelling to a subcutaneous injection with phytohemagglutinin) in the experimental nestlings compared to the control nestlings. In addition, the experimental nestlings showed a tendency for a reduced chance to be found breeding the following year. Furthermore, our results showed that in the experimental nestlings, which suffered from unfavourable growth conditions, tarsus length was inversely related to cell-mediated immunocompetence, whereas in control nestlings this relationship was significantly positive. The relationship between cell-mediated immunity and body condition was found to be significantly positive in the experimental nestlings while in control nestlings there was no relationship between both variables. The latter finding suggests different priorities of investment in body condition between different growth conditions. PMID- 15891822 TI - Will loss of snow cover during climatic warming expose New Zealand alpine plants to increased frost damage? AB - If snow cover in alpine environments were reduced through climatic warming, plants that are normally protected by snow-lie in winter would become exposed to greater extremes of temperature and solar radiation. We examined the annual course of frost resistance of species of native alpine plants from southern New Zealand that are normally buried in snowbanks over winter (Celmisia haastii and Celmisia prorepens) or in sheltered areas that may accumulate snow (Hebe odora) and other species, typical of more exposed areas, that are relatively snow-free (Celmisia viscosa, Poa colensoi, Dracophyllum muscoides). The frost resistance of these principal species was in accord with habitat: those from snowbanks or sheltered areas showed the least frost resistance, whereas species from exposed areas had greater frost resistance throughout the year. P. colensoi had the greatest frost resistance (-32.5 degrees C). All the principal species showed a rapid increase in frost resistance from summer to early winter (February-June) and maximum frost resistance in winter (July-August). The loss of resistance in late winter to early summer (August-December) was most rapid in P. colensoi and D. muscoides. Seasonal frost resistance of the principal species was more strongly related to daylength than to temperature, although all species except C. viscosa were significantly related to temperature when the influence of daylength was accounted for. Measurements of chlorophyll fluorescence indicated that photosynthetic efficiency of the principal species declined with increasing daylength. Levels of frost resistance of the six principal alpine plant species, and others measured during the growing season, were similar to those measured in tropical alpine areas and somewhat more resistant than those recorded in alpine areas of Europe. The potential for frost damage was greatest in spring. The current relationship of frost resistance with daylength is sufficient to prevent damage at any time of year. While warmer temperatures might lower frost resistance, they would also reduce the incidence of frosts, and the incidence of frost damage is unlikely to be altered. The relationship of frost resistance with daylength and temperature potentially provides a means of predicting the responses of alpine plants in response to global warming. PMID- 15891823 TI - Reproduction, foraging and the negative density-area relationship of a generalist rodent. AB - While many species show positive relationships between population density and habitat patch area, some species consistently show higher densities in smaller patches. Few studies have examined mechanisms that may cause species to have negative density-area relationships. We tested the hypothesis that greater reproduction in edge versus interior habitats and small versus large fragments contributes to higher densities of white-footed mice (Peromyscus leucopus) in small versus large forest fragments. We also examined vegetation structure and foraging tray utilization to evaluate if greater reproduction was a result of higher food availability. There were greater number of litters and proportion of females producing litters in the edge versus interior of forest fragments, which may have contributed to greater population growth rates and higher densities in edge versus interior and small versus large fragments. Data on vegetation structure and giving-up densities of seeds in artificial patches suggest that food availability may be higher in edge versus interior habitats and small versus large fragments. These results, in an area with few or no long-tailed weasels, provide a distinct contrast to the findings of Morris and Davidson (Ecology 81:2061, 2000) who observed lower reproduction in forest edge habitat as a result of high weasel predation, suggesting that specialist predators may be important in affecting the quality of edge habitat. While we cannot exclude the potential contributions of immigration, emigration, and mortality, our data suggest that greater reproduction in edge versus interior habitat is an important factor contributing to higher densities of P. leucopus in small fragments. PMID- 15891824 TI - Does the commonly used estimator of nutrient resorption in tree foliage actually measure what it claims to? AB - A descriptive temporal model is considered to be the best available estimator for accretion, resorption and proportional nutrient resorption. However, ecological studies rarely collect sufficient data for applying such a model. A less demanding and commonly used estimator for proportional resorption (PR) calculates PR as the percentage of the nutrient pool that is withdrawn from mature foliage before leaf abscission. Data from an intensive sampling campaign of the aboveground nutrient pools and fluxes of two Betula pendula Roth. stands were used. We showed that the commonly used estimator is not an accurate estimator for accretion, resorption and proportional resorption. The commonly used estimator underestimated the proportional resorption of N on the average by 3-10%, and the proportional resorption of P by 20-25%. The low accuracy of the estimations was shown to be caused by a lack of selectiveness of the commonly used estimator. In other words, the commonly used estimator does not measure the underlying processes in specific nutrient accretion and resorption at the stand level. However, when a sufficiently high sampling density with several samples at a given point in time is used, then the commonly used estimator preserves the ranking relationship between the PR of different sites for N in 97% of the cases and for P in 71%. The commonly used estimator can thus be used in comparative studies as an index for proportional nutrient resorption only. The quantitative results should not be taken literally, as they are based on only two sets of observations. However, the results show that the commonly used estimator should no longer be used as a measure for accretion, resorption or PR whenever the plant accretes nutrients in the foliage as a compensation for nutrient losses due to foliar leaching and litterfall during the growing season. PMID- 15891825 TI - Ecological implications of parasites in natural Daphnia populations. AB - In natural host populations, parasitism is considered to be omnipresent and to play an important role in shaping host life history and population dynamics. Here, we study parasitism in natural populations of the zooplankton host Daphnia magna investigating their individual and population level effects during a 2-year field study. Our results revealed a rich and highly prevalent community of parasites, with eight endoparasite species (four microsporidia, one amoeba, two bacteria and one nematode) and six epibionts (belonging to five different taxa: Chlorophyta, Bacillariophyceae, Ciliata, Fungi and Rotifera). Several of the endoparasites were associated with a severe overall fecundity reduction of the hosts, while such effects were not seen for epibionts. In particular, infections by Pasteuria ramosa, White Fat Cell Disease and Flabelliforma magnivora were strongly associated with a reduction in overall D. magna fecundity. Across the sampling period, average population fecundity of D. magna was negatively associated with overall infection intensity and total endoparasite richness. Population density of D. magna was negatively correlated to overall endoparasite prevalence and positively correlated with epibiont richness. Finally, the reduction in host fecundity caused by different parasite species was negatively correlated to both parasite prevalence and the length of the time period during which the parasite persisted in the host population. Consistent with epidemiological models, these results indicate that parasite mediated host damages influence the population dynamics of both hosts and parasites. PMID- 15891826 TI - Putting prey back together again: integrating predator-induced behavior, morphology, and life history. AB - The last decade has seen an explosion in the number of studies exploring predator induced plasticity. Recently, there has been a call for more comprehensive approaches that can identify functional relationships between traits, constraints on phenotypic responses, and the cost and benefits of alternative phenotypes. In this study, we exposed Helisoma trivolvis, a freshwater snail, to a factorial combination of three resource levels and five predator environments (no predator, one or two water bugs, and one or two crayfish) and examined ten traits including behavior, morphology, and life history. Each predator induced a unique suite of behavioral and morphological responses. Snails increased near-surface habitat use with crayfish but not with water bugs. Further, crayfish induced narrow and high shells whereas water bugs induced wide shells and wide apertures. In terms of life history, both predators induced delayed reproduction and greater mass at reproduction. However, crayfish induced a greater delay in reproduction that resulted in reduced fecundity whereas water bugs did not induce differences in fecundity. Resource levels impacted the morphology of H. trivolvis; snails reared with greater resource levels produced higher shells, narrower shells, and wider apertures. Resource levels also impacted snail life history; lower resources caused longer times to reproduction and reduced fecundity. Based on an analysis of phenotypic correlations, the morphological responses to each predator most likely represent phenotypic trade-offs. Snails could either produce invasion resistant shells for defense against water bugs or crush-resistant shells for defense against crayfish, but not both. Our use of a comprehensive approach to examine the responses of H. trivolvis has provided important information regarding the complexity of phenotypic responses to different environments, the patterns of phenotypic integration across environments, and the potential costs and benefits associated with plastic traits. PMID- 15891827 TI - Bumblebees, humble pollinators or assiduous invaders? A population comparison of foraging performance in Bombus terrestris. AB - Worldwide trade in non-native bumblebees remains largely unrestricted despite well-documented cases where introductions of non-native bees have gone dramatically wrong. Within Europe, indiscriminate importation of non-native populations of bumblebees (Bombus terrestris) for the pollination of glasshouse crops continues on a massive scale. However, no risk assessment has been conducted for these introductions, perhaps because B. terrestris is considered a native species, so shipping populations from one region to another has been implicitly assumed to present no risk. This view is clearly unjustified because Bombus terrestris populations differ significantly in their genetic makeup as demonstrated by strong differences in coat colour and behavioural traits. Therefore, for the first time we compare an important competitive trait, namely foraging performance, between commercially available B. terrestris populations in contrasting environments. We test whether commercially reared populations differ in their nectar foraging performance and whether this is influenced by both their source environment and the one they are introduced into. We do this by means of a reciprocal transplant experiment. Strong, consistent inter-population differences in performance occurred irrespective of test location: Canary Island bees (B. t. canariensis) were superior to Sardinian bees (B. t. sassaricus), which were generally superior to mainland European bees (B. t. terrestris). These inter population differences in performance were largely explained by inter-population variation in forager size, with larger bees being superior foragers. However, even when body size was accounted for, "native" bees were not superior to transplanted non-native bees in all but one case. We conclude that non-native populations, especially those with large foragers, can be highly competitive foragers. This could lead to their establishment and displacement of native bees. Therefore, we recommend that unregulated movements of non-native B. terrestris populations within Europe should not be carried out without a full risk assessment. PMID- 15891828 TI - Interactions between population processes in a cyclic species: parasites reduce autumn territorial behaviour of male red grouse. AB - The causes of population cycles fascinate and perplex ecologist. Most work have focused on single processes, whether extrinsic or intrinsic, more rarely on how different processes might interact to cause or mould the unstable population dynamics. In red grouse (Lagopus lagopus scoticus), two causal mechanisms have been supported: territorial behaviour (changes in autumn aggressiveness) and parasites (parasite induced reduction in fecundity). Here, we report on how these two regulatory processes might interact, by testing whether the parasite suspected to cause the grouse cycles, the nematode Trichostrongylus tenuis, reduces male autumn territorial behaviour. We either treated males with an anthelmintic, to remove parasites (dosed or D-males), or challenged them with infective T. tenuis larvae, to increase parasite intensity (challenged or C males). We first show that dosing was effective in removing T. tenuis parasites, while parasite intensities increased in challenged birds during the autumn. Because old males initially had more parasites than young males, the treatments generated greater differences in parasite intensity in old than in young males. We also show that various aspects of territorial behaviour (increase in testosterone-dependent comb size in autumn, territorial call rate, likelihood of winning territorial interactions and over-winter survival) were significantly higher in dosed than in challenged males, but in old birds only. Our data thus supported the hypothesis that parasites reduce male aggressiveness during the autumn territorial contests, and could thereby influence recruitment. Our results also highlight that the territorial behaviour of young males, which have fewer parasites, is not as limited by parasites as that of old, previously territorial males. We discuss the implications of these findings for our understanding of the processes regulating red grouse populations and causing their complex, unstable population dynamics. PMID- 15891829 TI - Precipitation pulse use by an invasive woody legume: the role of soil texture and pulse size. AB - Plant metabolic activity in arid and semi-arid environments is largely tied to episodic precipitation events or "pulses". The ability of plants to take up and utilize rain pulses during the growing season in these water-limited ecosystems is determined in part by pulse timing, intensity and amount, and by hydrological properties of the soil that translate precipitation into plant-available soil moisture. We assessed the sensitivity of an invasive woody plant, velvet mesquite (Prosopis velutina Woot.), to large (35 mm) and small (10 mm) isotopically labeled irrigation pulses on two contrasting soil textures (sandy-loam vs. loamy clay) in semi-desert grassland in southeastern Arizona, USA. Predawn leaf water potential (psi(pd)), the isotopic abundance of deuterium in stem water (deltaD), the abundance of 13C in soluble leaf sugar (delta13C), and percent volumetric soil water content (theta(v)) were measured prior to irrigation and repeatedly for 2 weeks following irrigation. Plant water potential and the percent of pulse water present in the stem xylem indicated that although mesquite trees on both coarse- and fine-textured soils quickly responded to the large irrigation pulse, the magnitude and duration of this response substantially differed between soil textures. After reaching a maximum 4 days after the irrigation, the fraction of pulse water in stem xylem decreased more rapidly on the loamy-clay soil than the sandy-loam soil. Similarly, on both soil textures mesquite significantly responded to the 10-mm pulse. However, the magnitude of this response was substantially greater for mesquite on the sandy-loam soil compared to loamy-clay soil. The relationship between psi(pd) and delta13C of leaf-soluble carbohydrates over the pulse period did not differ between plants at the two sites, indicating that differences in photosynthetic response of mesquite trees to the moisture pulses was a function of soil water availability within the rooting zone rather than differences in plant biochemical or physiological constraints. Patterns of resource acquisition by mesquite during the dynamic wetting-drying cycle following rainfall pulses is controlled by a complex interaction between pulse size and soil hydraulic properties. A better understanding of how this interaction affects plant water availability and photosynthetic response is needed to predict how grassland structure and function will respond to climate change. PMID- 15891830 TI - Multiple predator effects result in risk reduction for prey across multiple prey densities. AB - Investigating how prey density influences a prey's combined predation risk from multiple predator species is critical for understanding the widespread importance of multiple predator effects. We conducted experiments that crossed six treatments consisting of zero, one, or two predator species (hellgrammites, greenside darters, and creek chubs) with three treatments in which we varied the density of mayfly prey. None of the multiple predator effects in our system were independent, and instead, the presence of multiple predator species resulted in risk reduction for the prey across both multiple predator combinations and all three levels of prey density. Risk reduction is likely to have population-level consequences for the prey, resulting in larger prey populations than would be predicted if the effects of multiple predator species were independent. For one of the two multiple predator combinations, the magnitude of risk reduction marginally increased with prey density. As a result, models predicting the combined risk from multiple predator species in this system will sometimes need to account for prey density as a factor influencing per-capita prey death rates. PMID- 15891831 TI - Larval food limitation in butterflies: effects on adult resource allocation and fitness. AB - Allocation of larval food resources affects adult morphology and fitness in holometabolous insects. Here we explore the effects on adult morphology and female fitness of larval semi-starvation in the butterfly Speyeria mormonia. Using a split-brood design, food intake was reduced by approximately half during the last half of the last larval instar. Body mass and forewing length of resulting adults were smaller than those of control animals. Feeding treatment significantly altered the allometric relationship between mass and wing length for females but not males, such that body mass increased more steeply with wing length in stressed insects as compared to control insects. This may result in changes in female flight performance and cost. With regard to adult life history traits, male feeding treatment or mating number had no effect on female fecundity or survival, in agreement with expectations for this species. Potential fecundity decreased with decreasing body mass and relative fat content, but there was no independent effect of larval feeding treatment. Realized fecundity decreased with decreasing adult survival, and was not affected by body mass or larval feeding treatment. Adult survival was lower in insects subjected to larval semi starvation, with no effect of body mass. In contrast, previous laboratory studies on adult nectar restriction showed that adult survival was not affected by such stress, whereas fecundity was reduced in direct 11 proportion to the reduction of adult food. We thus see a direct impact of larval dietary restriction on survival, whereas fecundity is affected by adult dietary restriction, a pattern reminiscent of a survival/reproduction trade-off, but across a developmental boundary. The data, in combination with previous work, thus provide a picture of the intra-specific response of a suite of traits to ecological stress. PMID- 15891832 TI - The importance of microclimate variation in determining size, growth and survival of avian offspring: experimental evidence from a cavity nesting passerine. AB - Organisms are expected to balance energy allocation in such a way that fitness is maximized. While much research has focussed on allocation strategies of reproducing parents, in particular birds, relatively little attention has been paid to how nestlings allocate energy while in the nest. Nestling birds are faced with a trade-off between devoting energy to growth or to thermoregulation, and in altricial species it is likely that the thermal environment of the nest site influences the nature of this trade-off. Here, we experimentally investigate how altering the microclimate of nests affects the growth, size and survival, as well as cell-mediated immune (CMI) response, of nestling tree swallows (Tachycineta bicolor) in a temperate environment. We place air-activated heating pads in nests of swallows when young were between 4 days and 16 days of age, and compared performance of offspring to control nests. Our manipulation raised temperatures of heated nests by approximately 5 degrees C compared to control nests. Offspring in heated nests had enhanced survival while in the nest, and we also found that they were heavier and had longer ninth primary feathers at 16 days of age. In addition, heating nest boxes resulted in significantly faster growth of primaries, and there was a trend for growth rates of mass to also be higher in heated nests. There were no significant differences between heated and control nests in growth rate or size of tarsus at age 16 days, and we speculate that this lack of response to elevated nest temperatures may be due to growth of skeletal structures being limited by other factors such as calcium availability. We also found no difference between heated and control nests in CMI response. Nonetheless, our results show overall that increasing temperatures of nests has significant benefits that enhance the fitness of offspring. As provisioning rates to offspring did not differ between heated and control nests, we suspect that the beneficial effects of heating were not the consequence of changes in parental behaviour. Our results provide insight into factors, other than food supply, that have important consequences in determining reproductive success of birds breeding in temperate environments. PMID- 15891833 TI - Do lizards and snakes really differ in their ability to take large prey? A study of relative prey mass and feeding tactics in lizards. AB - Adaptations of snakes to overpower and ingest relatively large prey have attracted considerable research, whereas lizards generally are regarded as unable to subdue or ingest such large prey items. Our data challenge this assumption. On morphological grounds, most lizards lack the highly kinetic skulls that facilitate prey ingestion in macrostomate snakes, but (1) are capable of reducing large items into ingestible-sized pieces, and (2) have much larger heads relative to body length than do snakes. Thus, maximum ingestible prey size might be as high in some lizards as in snakes. Also, the willingness of lizards to tackle very large prey items may have been underestimated. Captive hatchling scincid lizards (Bassiana duperreyi) offered crickets of a range of relative prey masses (RPMs) attacked (and sometimes consumed parts of) crickets as large as or larger than their own body mass. RPM affected foraging responses: larger crickets were less likely to be attacked (especially on the abdomen), more likely to be avoided, and less likely to provide significant nutritional benefit to the predator. Nonetheless, lizards successfully attacked and consumed most crickets < or =35% of the predator's own body mass, representing RPM as high as for most prey taken by snakes. Thus, although lizards lack the impressive cranial kinesis or prey-subduction adaptations of snakes, at least some lizards are capable of overpowering and ingesting prey items as large as those consumed by snakes of similar body sizes. PMID- 15891834 TI - Scale-dependent mechanisms in the population dynamics of an insect herbivore. AB - A multiscale approach has lead to significant advances in the understanding of species population dynamics. The scale-dependent nature of population processes has been particularly clearly illustrated for insect herbivores. However, one of the most well-studied insect herbivores, the galling sawfly Euura lasiolepis, has to date been examined almost exclusively at fine spatial scales. The preference performance, plant vigour and larval survival hypotheses are well supported by this species. Here, we test these hypotheses at a spatial scale larger than that previously considered, i.e. across a landscape in northern Arizona represented by an altitudinal gradient encompassing a series of drainages. We also develop a qualitative model for understanding the population dynamics of E. lasiolepis based on patterns of survival and mortality found in this study and previous ones. Gall density was highly variable across the altitudinal gradient, not explained by host plant variables, and thus a poor surrogate for population abundance. These findings for the first time fail to support the plant vigour and preference hierarchy hypotheses for E. lasiolepis. Dispersal limitation most likely explains the lack of support for these hypotheses at this scale. By contrast, sawfly survival, gall abortion, parasitism and larval mortality were well explained by host plant quality variables and altitude. The larval survival hypothesis was well supported and is thus comparatively scale-invariant. A qualitative model developed here highlighted the importance of both willow water status and disturbance in determining host plant quality, as well as an apparent trade off between shoot length and plant moisture status in determining vital rates across the altitudinal gradient. This study thus demonstrated for the first time the scale-dependent nature of mechanisms underlying the population dynamics E. lasiolepis, and identified the interaction between parasitism and altitude as a novel mechanism underlying spatial patterns in the survival and mortality patterns of this species. PMID- 15891835 TI - Contribution analysis of body mass dynamics in Daphnia. AB - The concept of body mass dynamics can be viewed as part of life history theory, but its potential has remained largely untapped due to a lack of analytical methodology. We therefore propose a method, called contribution analysis, which enables us to decompose a change in body mass into contributions associated with variations in individual egg mass, clutch size, and standard somatic mass (somatic mass adjusted to body length). The advantage of contribution analysis is that various contributions are expressed in the same units (units of mass) and show the amount of resources committed to changes in the individual traits, while the traits themselves are measured in different units and thus incomparable on their own. The method is tuned to study zooplankton, and is applied to examine body mass dynamics in Daphnia galeata. We found that when recovering from a poor resource environment just above the threshold food concentration, Daphnia primarily increase their standard somatic mass, that is, restore body condition. When the trophic environment improves further but remains below the incipient limiting level, resources are invested equally to enhance body condition and reproduction in terms of clutch size. Finally, when food is no longer a limiting factor, almost all resources are committed to increase clutch size. While individual egg mass also varies, it never attracts more resources than the shift in the most prioritized trait. We suggest that the significance of this shift in resource allocation priorities is to keep an adult female alive in a poor environment and thus to allow her to retain her reproductive potential for better conditions in the future. Contribution analysis of body mass dynamics may allow us to detect flexible allocation strategies in a changing natural environment. PMID- 15891836 TI - A test of geographic assignment using isotope tracers in feathers of known origin. AB - We used feathers of known origin collected from across the breeding range of a migratory shorebird to test the use of isotope tracers for assigning breeding origins. We analyzed deltaD, delta13C, and delta15N in feathers from 75 mountain plover (Charadrius montanus) chicks sampled in 2001 and from 119 chicks sampled in 2002. We estimated parameters for continuous-response inverse regression models and for discrete-response Bayesian probability models from data for each year independently. We evaluated model predictions with both the training data and by using the alternate year as an independent test dataset. Our results provide weak support for modeling latitude and isotope values as monotonic functions of one another, especially when data are pooled over known sources of variation such as sample year or location. We were unable to make even qualitative statements, such as north versus south, about the likely origin of birds using both deltaD and delta13C in inverse regression models; results were no better than random assignment. Probability models provided better results and a more natural framework for the problem. Correct assignment rates were highest when considering all three isotopes in the probability framework, but the use of even a single isotope was better than random assignment. The method appears relatively robust to temporal effects and is most sensitive to the isotope discrimination gradients over which samples are taken. We offer that the problem of using isotope tracers to infer geographic origin is best framed as one of assignment, rather than prediction. PMID- 15891837 TI - Phenotypic and genetic differentiation between native and introduced plant populations. AB - Plant invasions often involve rapid evolutionary change. Founder effects, hybridization, and adaptation to novel environments cause genetic differentiation between native and introduced populations and may contribute to the success of invaders. An influential idea in this context has been the Evolution of Increased Competitive Ability (EICA) hypothesis. It proposes that after enemy release plants rapidly evolve to be less defended but more competitive, thereby increasing plant vigour in introduced populations. To detect evolutionary change in invaders, comparative studies of native versus introduced populations are needed. Here, we review the current empirical evidence from: (1) comparisons of phenotypic variation in natural populations; (2) comparisons of molecular variation with neutral genetic markers; (3) comparisons of quantitative genetic variation in a common environment; and (4) comparisons of phenotypic plasticity across different environments. Field data suggest that increased vigour and reduced herbivory are common in introduced plant populations. In molecular studies, the genetic diversity of introduced populations was not consistently different from that of native populations. Multiple introductions of invasive plants appear to be the rule rather than the exception. In tests of the EICA hypothesis in a common environment, several found increased growth or decreased resistance in introduced populations. However, few provided a full test of the EICA hypothesis by addressing growth and defence in the same species. Overall, there is reasonable empirical evidence to suggest that genetic differentiation through rapid evolutionary change is important in plant invasions. We discuss conceptual and methodological issues associated with cross-continental comparisons and make recommendations for future research. When testing for EICA, greater emphasis should be put on competitive ability and plant tolerance. Moreover, it is important to address evolutionary change in characteristics other than defence and growth that could play a role in plant invasions. PMID- 15891838 TI - 'Are fish what they eat' all year round? AB - Isotope turnover in muscle of ectotherms depends primarily on growth rather than on metabolic replacement. Ectotherms, such as fish, have a discontinuous pattern of growth over the year, so the isotopic signature of muscle (delta13C and delta15N) may only reflect food consumed during periods of growth. In contrast, the liver is a regulatory tissue, with a continuous protein turnover. Therefore, the isotopic composition of liver should respond year round to changes in the isotopic signature of food sources. Therefore, we predicted that (1) Whitefish in Lake Geneva would have larger seasonal variation in the isotopic variation of the liver compared to that of the muscle tissue, and (2) the isotope composition of fish muscle would reflect a long-term image of the isotope composition of the food consumed only throughout the growth period. To test these expectations, we compared the isotope compositions of Whitefish muscle, liver and food in a 20 month study. We found that the seasonal amplitude of isotope variation was two to three times higher in liver compared to muscle tissue. During the autumn and winter, when growth was limited, only the isotopic signature of liver responded to changes in the isotope composition of the food sources. The delta13C and delta15N of muscle tissue only reflected the food consumed during the spring and summer growth period. PMID- 15891839 TI - Photosynthesis and reflectance indices for rainforest species in ecosystems undergoing progression and retrogression along a soil fertility chronosequence in New Zealand. AB - Measurements of photosynthesis at saturating irradiance and CO2 partial pressure, Amax, "adjusted" normalised difference vegetation index, RaNDVI, and photochemical reflectance index, RPRI, were made on trees sampled along a soil chronosequence to investigate the relationship between carbon uptake and ecosystem development in relation to nutrient availability. Measurements were made on the three most dominant species at six sites along the sequence in South Westland, New Zealand with soil age ranging from < 6 to 120,000 years resulting from the retreat of the Franz Josef glacier. The decrease in soil phosphorus availability with increasing soil age and high soil nitrogen availability at the two youngest sites, due to the presence of a nitrogen-fixing species, provided marked differences in nutrient availability. Mean Amax was high at the two youngest sites, then decreased markedly with increasing site age. Analysis of the data for individual species within sites revealed separation of groups of species in the response of Amax to Nm and Pm, suggesting complex interactions between the two nutrients. There were strong linear relationships for leaf-level RaNDVI and RPRI with Amax, at high irradiance, showing that measurements of reflectance indices can be used to estimate Amax for foliage with a range in morphology and nutrient concentrations. Notwithstanding the change in species composition from angiosperms to conifers with increasing site age, the presence of nitrogen-fixing species, the variability in foliage morphology from flat leaves to imbricate scales and a wide range in foliar nitrogen and phosphorus concentrations, there were strong positive linear relationships between site average Amax and foliage nitrogen, Nm, and phosphorus, Pm, concentrations on a foliage mass basis. The results provide insights to interpret the regulation of photosynthesis across natural ecosystems with marked gradients in nitrogen and phosphorus availability. PMID- 15891840 TI - The effect of cold-induced increased metabolic rate on the rate of 13C and 15N incorporation in house sparrows (Passer domesticus). AB - Animals with high metabolic rates are believed to have high rates of carbon and nitrogen isotopic incorporation. We hypothesized that (1) chronic exposure to cold, and hence an increase in metabolic rate, would increase the rate of isotopic incorporation of both 13C and 15N into red blood cells; and (2) that the rate of isotopic incorporation into red blood cells would be allometrically related to body mass. Two groups of sparrows were chronically exposed to either 5 or 22 degrees C and switched from a 13C-depleted C3-plant diet to a more 13C enriched C4-plant one. We used respirometry to estimate the resting metabolic rate (VO2) of birds exposed chronically to our two experimental temperatures. The allometric relationship between the rate of 13C incorporation into blood and body mass was determined from published data. The (VO2) of birds at 5 degrees C was 1.9 times higher than that of birds at 22 degrees C. Chronic exposure to a low temperature did not have an effect on the rate of isotopic incorporation of 15N save for a very small effect on the incorporation of 13C. The isotopic incorporation rate of 13C was 1.5 times faster than that of 15N. The fractional rate of 13C incorporation into avian blood was allometrically related to body mass with an exponent similar to -1/4. We conclude that the relationship between metabolic rate and the rate of isotopic incorporation into an animal's tissues is indirect. It is probably mediated by protein turnover and thus more complex than previous studies have assumed. PMID- 15891841 TI - Ecophysiology of first and second generation hybrids in a natural plant hybrid zone. AB - Hybrids between related species vary widely in relative fitness, and that fitness can depend upon the environment. We investigated aspects of physiology that might influence fitness patterns in a plant hybrid zone. Seeds of Ipomopsis aggregata, I. tenuituba, F1 hybrids, F2 hybrids, and offspring of crosses between natural hybrids were planted into the relatively mesic site of origin for I. aggregata and the drier site for natural hybrids. We measured rates of photosynthesis (Amax), transpiration (E), instantaneous (A/E) and long-term (delta13C) indices of water use efficiency (WUE), and leaf nitrogen and carbon. We also examined correlations of these traits with plant size. Photosynthetic rate and A/E were higher in vegetative than flowering plants. WUE varied between sites and years, but differences among genotypic classes were spatially and temporally consistent. Instantaneous WUE was higher for F1 hybrids than for the average of the parental species, thereby showing heterosis. There was no evidence of hybrid breakdown, as WUE was no different in the F2 than the average across the F1 and parental species. Nor did WUE depend on cross direction in producing F1 progeny. Carbon isotope discrimination revealed higher long-term water use efficiency in I. tenuituba than I. aggregata. Leaf nitrogen was higher in I. tenuituba than I. aggregata, and higher in offspring of natural hybrids than in the F2. Results indicate heterosis for water use efficiency, with no hybrid breakdown. Heterosis in WUE may help to explain the relatively high survival of both reciprocal F1 hybrids in dry sites within the natural hybrid zone. PMID- 15891842 TI - Scaling from plot experiments to landscapes: studying grasshoppers to inform forest ecosystem management. AB - Ecologists studying food web interactions routinely conduct their experiments at scales of 1-10 m(2) whereas real-world landscape-level management problems exist on scales of 10(6) m(2) or larger. It is often asserted that the experimental tradition in ecology has little to offer to environmental management because small scale empirical insights are not easily, if at all, translatable to the large scale problems. Small scale experiments are very local in nature and they are conducted in ways that tend to homogenize background environmental variation. Real world management is conducted across vast landscapes. Managers routinely must wrestle with complexity that is introduced by the heterogeneous structure of those landscapes and they often have limited recourse to do careful experimentation. How then is empirical ecological science ever to inform landscape-level management? The solution to this dilemma lies in arriving at good working conceptualizations of ecosystem structure and function that embody principles that are relatively scale independent. In this paper, the evolutionary ecological principle of foraging versus predation risk avoidance trade-offs is proffered as one central organizing conceptualization for plant-herbivore interactions across all systems. The utility of this conceptualization is first illustrated by presenting results of detailed experiments involving spider predators, grasshopper herbivores, and two classes of plant resources that afford grasshoppers differential protection from predators: nutritionally superior but risky grasses and less nutritious but safer herbs. The paper then shows how the foraging versus predation risk avoidance conceptualization in the context of a "landscape of fear" can be applied to manage large herbivore impacts of forest regeneration following forest harvesting. I present results of landscape-scale experiments that mediate predation risk of the herbivores through manipulation of safe habitat in order to enlist herbivores to facilitate boreal forest mixed species regeneration through preferential foraging of certain woody species. PMID- 15891843 TI - Dimensional approaches to designing better experimental ecosystems: a practitioners guide with examples. AB - Enclosed, experimental ecosystems ("mesocosms") are now widely used research tools in ecology. However, the small size, short duration and often simplified biological and physical complexity of mesocosm experiments raises questions about extrapolating results from these miniaturized ecosystems to nature. Dimensional analysis, a technique widely used in engineering to create scale models, employs "compensatory distortion" as a means of maintaining functional similarity in properties and relationships of interest. An earlier paper outlined a general approach to applying dimensional analysis to the construction and interpretation of mesocosm experiments (Petersen and Hastings in Am Nat 157:324, 2001). In this paper we use examples, largely drawn from the aquatic literature, to illustrate how dimensional approaches might be used to maintain key ecological properties. Such key properties include effective habitat size, environmental variability, vertical and horizontal gradients, and interactions among habitats. We distinguish both continuous and discrete approaches that can be used to achieve functional similarity through compensatory distortion. In addition to its potential as a tool for improving the realism of experimental ecosystems, the dimensional approach points towards new options for developing, testing and advancing our understanding of scaling relationships in nature. PMID- 15891844 TI - Do local processes scale to global patterns? The role of drought and the species pool in determining treehole insect diversity. AB - Global patterns in community species richness may represent limitations operating at the local scale, such as competitive exclusion and habitat suitability, or, in the case of unsaturated communities, limitations to the species pool of biogeographic regions. Separating the effects of local and regional processes on community richness requires combining small-scale experiments with broad surveys. I examine limitations to the number of aquatic insect species per treehole at the scale of a single woodland, and between eight biogeographic regions. Variation in species richness at the woodland scale can largely be explained by small-scale differences between treeholes in drought disturbance, as shown for British treehole insects for 2 years. At the global scale, drought disturbance is a relatively poor predictor of patterns in the local richness of treehole mosquitoes. Instead, regional differences in the species pool explain most of the variation in local mosquito richness. Treeholes in at least the most species-poor regions appear to be unsaturated with mosquito species. In this system, therefore, local processes do not necessarily scale to global patterns. PMID- 15891845 TI - Variation in plant quality and the population dynamics of herbivores: there is nothing average about aphids. AB - In the attempt to use results from small-scale studies to make large-scale predictions, it is critical that we take into account the greater spatial heterogeneity encountered at larger spatial scales. An important component of this heterogeneity is variation in plant quality, which can have a profound influence on herbivore population dynamics. This influence is particularly relevant when we consider that the strength of density dependence can vary among host plants and that the strength of density dependence determines the difference between exponential and density- dependent growth. Here, we present some simple models and analyses designed to examine the impact of variable plant quality on the dynamics of insect herbivore populations, and specifically the consequences of variation in the strength of density dependence among host plants. We show that average values of herbivore population growth parameters, calculated from plants that vary in quality, do not predict overall population growth. Furthermore, we illustrate that the quality of a few individual plants within a larger plant population can dominate herbivore population growth. Our results demonstrate that ignoring spatial heterogeneity that exists in herbivore population growth on plants that differ in quality can lead to a misunderstanding of the mechanisms that underlie population dynamics. PMID- 15891846 TI - Scaling up from local competition to regional coexistence across two scales of spatial heterogeneity: insect larvae in the fruits of Apeiba membranacea. AB - Species that live in patchy and ephemeral habitats can compete strongly for resources within patches at a small scale. The ramifications of these interactions for population dynamics and coexistence at regional scales will depend on the intraspecific and interspecific distributions of individuals among patches. Spatial heterogeneity due to independent aggregation of competitors among patchy habitats is an important mechanism maintaining species diversity. I describe regional patterns of aggregation for four species of insect larvae in the fruits of Apeiba membranacea, a Neotropical rainforest tree. This aggregation results from variation in densities at a small scale (among the fruits under a single tree), compounded by significant variation among trees in both mean densities and degrees of aggregation. Both the degrees of aggregation and mean densities are statistically independent within and across species at both spatial scales. I evaluate the regional consequences of these spatial patterns by using maximum likelihood methods to parameterize a model that includes both explicit measures of the strength of competition and spatial variation at both within- and among-tree spatial scales. Despite strong competitive interactions among these species, during 2 years the observed spatial variation at both scales combined was sufficient to explain the coexistence of these species, although other coexistence mechanisms may also operate simultaneously. The observed spatial variation at small spatial scales may not be sufficient for coexistence, indicating the importance of considering multiple sources of spatial heterogeneity when scaling up from experiments that investigate local interactions to regional patterns of coexistence. PMID- 15891847 TI - Scaling up population dynamics: integrating theory and data. AB - How to scale up from local-scale interactions to regional-scale dynamics is a critical issue in field ecology. We show how to implement a systematic approach to the problem of scaling up, using scale transition theory. Scale transition theory shows that dynamics on larger spatial scales differ from predictions based on the local dynamics alone because of an interaction between local-scale nonlinear dynamics and spatial variation in density or the environment. Based on this theory, a systematic approach to scaling up has four steps: (1) derive a model to translate the effects of local dynamics to the regional scale, and to identify key interactions between nonlinearity and spatial variation, (2) measure local-scale model parameters to determine nonlinearities at local scales, (3) measure spatial variation, and (4) combine nonlinearity and variation measures to obtain the scale transition. We illustrate the approach, with an example from benthic stream ecology of caddisflies living in riffles. By sampling from a simulated system, we show how collecting the appropriate data at local (riffle) scales to measure nonlinearities, combined with measures of spatial variation, leads to the correct inference for dynamics at the larger scale of the stream. The approach provides a way to investigate the mechanisms and consequences of changes in population dynamics with spatial scale using a relatively small amount of field data. PMID- 15891848 TI - Large-scale questions and small-scale data: empirical and theoretical methods for scaling up in ecology. PMID- 15891849 TI - The role of predation in the decline and extirpation of woodland caribou. AB - To select appropriate recovery strategies for endangered populations, we must understand the dynamics of small populations and distinguish between the possible causes that drive such populations to low numbers. It has been suggested that the pattern of population decline may be inversely density-dependent with population growth rates decreasing as populations become very small; however, empirical evidence of such accelerated declines at low densities is rare. Here we analyzed the pattern of decline of a threatened population of woodland caribou (Rangifer tarandus caribou) in British Columbia, Canada. Using information on the instantaneous rate of increase relative to caribou density in suitable winter foraging habitat, as well as on pregnancy rates and on causes and temporal distribution of mortalities from a sample of 349 radiocollared animals from 15 subpopulations, we tested 3 hypothesized causes of decline: (a) food regulation caused by loss of suitable winter foraging habitat, (b) predation-sensitive foraging caused by loss of suitable winter foraging habitat and (c) predation with caribou being secondary prey. Population sizes of caribou subpopulations ranged from < 5 to > 500 individuals. Our results showed that the rates of increase of these subpopulations varied from -0.1871 to 0.0496 with smaller subpopulations declining faster than larger subpopulations. Rates of increase were positively related to the density of caribou in suitable winter foraging habitat. Pregnancy rates averaged 92.4% +/-2.24 and did not differ among subpopulations. In addition, we found predation to be the primary cause of mortality in 11 of 13 subpopulations with known causes of mortality and predation predominantly occurred during summer. These results are consistent with predictions that caribou subpopulations are declining as a consequence of increased predation. Recovery of these woodland caribou will thus require a multispecies perspective and an appreciation for the influence of inverse density dependence on population trajectories. PMID- 15891850 TI - Experimental evaluation and thermo-physical analysis of thermogenesis in male and female cycad cones. AB - Thermogenically elevated cone temperatures were measured in two Macrozamia cycad species that differ in their daily heating time. Mathematical models of the cones' thermo-physics were tested for their accuracy in predicting these cone temperatures and for comparison of the energetics of both species and the sexes within species. These models accurately predicted temperatures over approximately 8-h periods with average errors of: 0.46 degrees C for Macrozamia lucida, pollinated by the thrips, Cycadothrips chadwicki, that moves during mid-day concurrent with cone heating; and 0.38 degrees C for Macrozamia machinii, pollinated by the weevil, Tranes sp., that moves after sunset during cone heating. The combination of models and experiments revealed a thermogenic sexual dimorphism in both species. For M. lucida, the estimated female mass specific metabolisms, and their theoretically possible and actual temperature increases due to thermogenic metabolism were only 57, 67, and 76% of males. In addition, female thermogenic metabolisms began and peaked much earlier and lasted significantly longer than males (all differences >1 h), and female metabolic peaks preceded their temperature peaks by 65 vs. 46 min for males. The timing of almost all male cone metabolic peaks was optimized with respect to the diurnal ambient heating cycle so that cone temperatures achieved a maximum temperature gain, whereas most female metabolic peaks occurred much earlier than optimal. In M. machinii, thermogenic sexual dimorphism is much larger since its male peak metabolisms are larger, and its females' peaks are much smaller compared to those of M. lucida. This study provides new information regarding the energetics of cycad cones that is relevant to understanding the interactions of the plant traits with their obligate pollinators' behavior. PMID- 15891851 TI - Fish track wastewater pollution to estuaries. AB - Excess nitrogen is a forceful agent of ecological change in coastal waters, and wastewater is a prominent source of nitrogen. In catchments where multiple sources of nitrogen pollution co-exist, biological indicators are needed to gauge the degree to which wastewater-N can propagate through the receiving food webs. The purpose of this study was to test whether estuarine fish are suitable as indicators of sewage-N pollution. Fish were analysed from three estuaries within a 100-km strip on the Australian East Coast. The estuaries differ substantially in wastewater loading: (1) the Maroochy Estuary receives a large fraction of the local shire's treated sewage, (2) the Mooloolah Estuary has no licensed treated wastewater outfalls but marinas/harbours and storm-water may contribute nitrogen, and (3) the Noosa Estuary which neither receives licensed discharges nor has suspected wastewater loads. Sampling for fish included both high rainfall ('wet' season) and low rainfall ('dry' season) periods. Muscle-delta15N was the variable predicted to respond to treated wastewater loading, reflecting the relative enrichment in 15N resulting from the treatment process and distinguishing it from alternative N sources such as fertiliser and natural nitrogen inputs (both 15N depleted). Of the 19 fish species occurring in all three estuaries, those from the Maroochy Estuary had significantly elevated delta15N values (up to 9.9 per thousand), and inter-estuarine differences in fish-delta15N were consistent across seasons. Furthermore, not only did all fish from the estuary receiving treated wastewater carry a very distinctive sewage-N tissue signal, but enriched muscle-delta15N was also evident in all species sampled from the one estuary in which sewage contamination was previously only suspected (i.e. the Mooloolah Estuary: 0.2-4.8 per thousand enrichment over fish from reference system). Thus, fish-delta15N is a suitable indicator of wastewater-N not only in systems that receive large loads, but also for the detection of more subtle nitrogen inputs. Arguably, fish may be preferred indicators of sewage-N contamination because they: (1) integrate nitrogen inputs over long time periods, (2) have an element of 'ecological relevance' because fish muscle-delta15N reflect movement of sewage N through the food chain, and (3) pollution assessments can usually be based on evidence from multiple species. PMID- 15891852 TI - Clustered root distribution in mature stands of Fagus sylvatica and Picea abies. AB - Distribution of small roots (diameter between 2 mm and 5 mm) was studied in 19 pits with a total of 72 m(2) trench profile walls in pure stands of Fagus sylvatica and Picea abies. Root positions within the walls were marked and transformed into x-coordinates and y-coordinates. In a GIS-based evaluation, zones of potential influence around each root were calculated. The total potential influence produced isoline maps of relative root influence zones, thus indicating small root clustering. The questions studied were (1) whether there were marked clusters of small roots in the soil and (2) whether trees surrounding the pit (defined as tree density) correlate with the root abundance and distribution on the trench profile walls. Small roots of both species showed maximum abundance in the top 20 cm of the soil, where pronounced root clusters occurred next to areas with only low root accumulation. The area of root clusters did not differ significantly between the two stands. Weighted clumping, WC, calculated as a product of root class, and its area was used as an index of root clustering, which again did not differ between beech and spruce stands. However, evaluations on a single root level showed that beech achieved the same degree of clustering with lower number of roots. Regardless of soil properties related to root clusters, a significantly higher clustering acquired per root for beech than for spruce suggests beech to be more efficient in belowground acquisition of space. Because none of the parameters describing root clustering were correlated with tree density around the investigated soil profiles, clusters of small roots are inherently present within the tree stands. PMID- 15891853 TI - Marine-derived nitrogen and carbon in freshwater-riparian food webs of the Copper River Delta, southcentral Alaska. AB - After rearing to adulthood at sea, coho salmon (Oncorhynchus kisutch) return to freshwater to spawn once and then die on or near their spawning grounds. We tested the hypothesis that spawning coho salmon return marine N and C to beaver (Castor canadensis) ponds of the Copper River Delta (CRD), Cordova, southcentral Alaska, thereby enhancing productivity of the aquatic food webs that support juvenile coho salmon. We sampled three types of pond treatment: (1) natural enrichment by spawning salmon, (2) artificial enrichment via addition of salmon carcasses and eggs, and (3) ponds with no salmon enrichment. All ponds supported juvenile coho salmon. Seasonal samples of stable isotopes revealed that juvenile coho salmon, threespine sticklebacks (Gasterosteus aculeatus), caddisfly larvae, leeches, and chironomid midge larvae were enriched with marine N and C. The aquatic vascular plants bur reed (Sparganium hyperboreum), pondweed (Potamogeton gramineus), and mare's tail (Hippuris vulgaris) were enriched with marine N only. Riparian vegetation (Sitka alder Alnus viridis ssp. sinuata and willow Salix spp.) did not show enrichment. Artificial additions of adult carcasses and eggs of coho salmon increased the delta15N and delta13C values of juvenile coho salmon. In this dynamic and hydrologically complex coastal environment, spawning coho salmon contributed marine N and C comprising 10-50% of the dietary needs of juvenile coho salmon through direct consumption of eggs and carcass material. Invertebrates that have assimilated marine N and C yield a further indirect contribution. This perennial subsidy maintains the productivity of the ecosystem of the coho salmon on the CRD. PMID- 15891854 TI - Foliar 15N natural abundance indicates phosphorus limitation of bog species. AB - Foliar delta15N, %N and %P in the dominant woody and herbaceous species across nutrient gradients in New Zealand restiad (family Restionaceae) raised bogs revealed marked differences in plant delta15N correlations with P. The two heath shrubs, Leptospermum scoparium (Myrtaceae) and Dracophyllum scoparium (Epacridaceae), showed considerable isotopic variation (-2.03 to -15.55 per thousand, and -0.39 to -12.06 per thousand, respectively) across the bogs, with foliar delta15N strongly and positively correlated with P concentrations in foliage and peat, and negatively correlated with foliar N:P ratios. For L. scoparium, the isotopic gradient was not linked to ectomycorrhizal (ECM) fractionation as ECMs occurred only on higher nutrient marginal peats where 15N depletion was least. In strong contrast, restiad species (Empodisma minus Sporadanthus ferrugineus, S. traversii) showed little isotopic variation across the same nutrient gradients. Empodisma minus and S. traversii had delta15N levels consistently around 0 per thousand (means of -0.12 per thousand and +0.15 per thousand respectively), and S. ferrugineus, which co-habited with E. minus, was more depleted (mean -4.97 per thousand). The isotopic differences between heath shrubs and restiads were similar in floristically dissimilar bogs and may be linked to contrasting nutrient demands, acquisition mechanisms, and root morphology. Leptospermum scoparium shrubs on low nutrient peats were stunted, with low tissue P concentrations, and high N:P ratios, suggesting they were P limited, which was probably exacerbated by markedly reduced mycorrhizal colonizations. The coupling of delta(15)N depletion and %P in heath shrubs suggests that N fractionation is promoted by P limitation. In contrast, the constancy in delta15N of the restiad species through the N and P gradients suggests that these are not suffering from P limitation. PMID- 15891855 TI - Examining the strength and possible causes of the relationship between fire history and Sudden Oak Death. AB - Fire can be a dominant process in the ecology of forest vegetation and can also affect forest disease dynamics. Little is known about the relationship between fire and an emerging disease epidemic called Sudden Oak Death, which is caused by a new pathogen, Phytophthora ramorum. This disease has spread across a large, fire-prone portion of California, killing great numbers of oaks and tanoaks and infecting most associated woody plants. Suitable hosts cover a much broader geographic range, raising concern over where the disease may spread. To understand the strength and potential sensitivities of a fire-disease relationship, we examined geographic patterns of confirmed P. ramorum infections in relation to past fire history. We found these infections to be extremely rare within the perimeter of any area burned since 1950. This finding is not caused by spatial bias in sampling for the disease, and is robust to variation in host abundance scenarios and to aggregation of closely spaced sampling locations. We therefore investigated known fire-related factors that could result in significantly lower incidence of the disease in relatively recently burned landscapes. Chemical trends in post-fire environments can influence the success of pathogens like P. ramorum, either by increasing plant nutrient stress or by reducing the occurrence of chemicals antagonistic to Phytophthoras. Succession in the absence of fire leads to greater abundance of host species, which will provide increased habitat for P. ramorum; this will also increase intraspecific competition where these trees are abundant, and other density-dependent effects (e.g. shading) can reduce resource allocation to defenses. Despite these findings about a fire-disease relationship, a much deeper understanding is necessary before fire can be actively used as a tool in slowing the epidemic. PMID- 15891856 TI - Altered parasite assemblages in raccoons in response to manipulated resource availability. AB - The role that host aggregation plays in structuring parasite assemblages was examined by experimentally increasing the contact rates of raccoons, Procyon lotor. Two populations of raccoons in southern New York were monitored for 2 years to determine baseline levels of host interaction and to identify the parasite assemblage. In the third year of the study, one population was provisioned with the addition of clumped food resources, while the other was provisioned with equal quantities of dispersed food resources. Remote photography showed that raccoons aggregated at clumped resources but not at dispersed resources, and therefore contact rates between individuals were higher in the site with clumped resources. There were no differences in parasitism between the sites prior to resource augmentation. Among ectoparasites, there were no significant changes in the prevalence or abundance of any species in response to the perturbation. In contrast, across the endoparasite assemblage within and across hosts, the prevalence of infection increased as a result of increased host contact. Strong increases in the prevalence of a few directly transmitted species and slight increases among most species lead to increased evenness in parasite prevalence, suggesting that parasites in this system are transmission limited. In addition, the number of parasite species per host (the parasite infracommunity) was higher in the clumped-resource population. These endoparasite results suggest that intraspecific variation in the species richness of parasite communities of individual hosts, and the prevalence of parasitic species in host populations as assessed across entire parasitic assemblages, is robustly influenced by intraspecific variation in the degree of host contact. Further, these results suggest that anthropogenic changes which alter resource availability may have important consequences for disease transmission in wildlife. PMID- 15891857 TI - Community assembly at the patch scale in a species rich tropical river. AB - In tropical floodplain rivers, communities associated with structurally complex habitats are disassembled and reassembled as aquatic organisms repeatedly colonize new areas in response to gradual but continuous changes in water level. Thus, a neutral model reflecting random colonization and extinction dynamics may be sufficient to predict assemblage patterns at the scale of local habitat patches. If water level fluctuations and associated patch dynamics are sufficiently predictable, however, community assembly on habitat patches also may be influenced by species-specific responses to habitat features and/or species interactions. We experimentally manipulated structural complexity and proximity to source habitat (which influences colonization rate) of simulated rocky patches in the littoral zone of a tropical lowland river and demonstrate significant effects of both factors on species density of fishes and macroinvertebrates. Interspecific variation in vagility significantly affected assemblage response to habitat complexity. In a second experiment, created habitat patches were sampled over time intervals ranging from 1 day to 36 days to examine temporal dynamics of community assembly. A null-model test revealed that assemblage structure became increasingly non-random, concomitant with increasing species density, over time. Community dynamics in newly formed habitat patches appeared to be dominated by dispersal, whereas in older patches, abundances of individual species increasingly were influenced by habitat characteristics. These data suggest that species-specific responses to environmental variation resulted, in part, because of species interactions. We conclude that community assembly in shallow habitats of this tropical lowland river is influenced by physical habitat characteristics, the spatial distribution of habitat patches, and species interactions as habitats are saturated with individuals. PMID- 15891858 TI - Evidence for mutualist limitation: the impacts of conspecific density on the mycorrhizal inoculum potential of woodland soils. AB - The ability of seedlings to establish can depend on the availability of appropriate mycorrhizal fungal inoculum. The possibility that mycorrhizal mutualists limit the distribution of seedlings may depend on the prevalence of the plant hosts that form the same type of mycorrhizal association as the target seedling species and thus provide inoculum. We tested this hypothesis by measuring ectomycorrhizal (EM) fine root distribution and conducting an EM inoculum potential bioassay along a gradient of EM host density in a pinyon juniper woodland where pinyon is the only EM fungal host while juniper and other plant species are hosts for arbuscular mycorrhizal (AM) fungi. We found that pinyon fine roots were significantly less abundant than juniper roots both in areas dominated aboveground by juniper and in areas where pinyon and juniper were co-dominant. Pinyon seedlings establishing in pinyon-juniper zones are thus more likely to encounter AM than EM fungi. Our bioassay confirmed this result. Pinyon seedlings were six times less likely to be colonized by EM fungi when grown in soil from juniper-dominated zones than in soil from either pinyon-juniper or pinyon zones. Levels of EM colonization were also reduced in seedlings grown in juniper-zone soil. Preliminary analyses indicate that EM community composition varied among sites. These results are important because recent droughts have caused massive mortality of mature pinyons resulting in a shift towards juniper dominated stands. Lack of EM inoculum in these stands could reduce the ability of pinyon seedlings to re-colonize sites of high pinyon mortality, leading to long term vegetation shifts. PMID- 15891859 TI - Rodent seed predation promotes differential recruitment among bird-dispersed trees in temperate secondary forests. AB - We investigated the role of seed predation by rodents in the recruitment of the fleshy-fruited trees Taxus baccata, Ilex aquifolium and Crataegus monogyna in temperate secondary forests in NW Spain. We measured the densities of dispersed seeds, early emerged seedlings, established recruits and adults, at four sites over a period of 2 years. Seed predation among species was compared by seed removal experiments and analysis of rodent larder-hoards. The three species differed markedly in local regeneration patterns. The rank order in the seed rain following decreasing seed density was Ilex, Taxus and Crataegus. However, Crataegus established 3.3 times more seedlings than Taxus. For all species, there was a positive linear relationship between the density of emerged seedlings and seed density, suggesting that recruitment was seed- rather than microsite limited. A consistent pattern of seed selection among species was exerted by rodents, which preferred Taxus and, secondarily, Ilex seeds to Crataegus seeds. Predation ranking was the inverse of that of seed protection against predators, measured as the mass of woody coat per mass unit of the edible fraction. Recruitment potential, evaluated as the ratio of seedlings to seeds, was negatively related to seed predation, with the rank order Crataegus > Ilex > Taxus. The selective early recruitment limitation exerted by predation may have a demographic effect in the long term, as judged by the positive relationship between early seedling emergence and the density of established recruits. By modulating the pre-emptive competition for seed safe sites, rodents may preclude the progressive exclusion of species that produce low numbers of seeds (i.e. Crataegus) by those dominant in seed number (i.e. Ilex, Taxus), or at least foster the evenness for site occupation among seedlings of different species. PMID- 15891860 TI - Characterising spatial and temporal variation in the finite rate of population increase across the northern range boundary of the annual grass Vulpia fasciculata. AB - Understanding the factors that influence plant distributions is a considerable challenge for ecologists in the face of environmental change. Here, we quantify spatial and temporal variation in the finite rate of population increase of the annual grass Vulpia fasciculata. Specifically, we test the hypothesis that the northern range boundary is associated with finite rates of population increase of less than one. Seeds of three ecotypes of the annual grass V. fasciculata were introduced annually across a range of sites in Great Britain both within (11) and to the north (4) of its current range boundary in each of 4 years. Populations failed to establish at 17% of target sites due to disturbance. At the remaining target sites, the finite rate of population increase, lambda, varied from 0.06 to 33.3 with a geometric mean of 1.88. Of the total variance in the rate of population growth, site and year effects accounted independently for 40% of the variation and in interaction for 50%; ecotype accounted for less than 5% of the variation. Variation in the weather between sites and years had little impact on plant performance, and there was no indication that the rate of population growth was lower to the north of the current range boundary. We conclude that current climatic conditions on the coast of Great Britain are not limiting the distribution of V. fasciculata and that seeds from across its current range have roughly equivalent colonising potential. PMID- 15891861 TI - Mask ventilation and cardiogenic pulmonary edema: "another brick in the wall". PMID- 15891862 TI - Illness severity and parental permission for clinical research in a pediatric ICU population. AB - OBJECTIVE: Research in child subjects requires parental permission. We examined whether parental authorization of involvement in a clinical study is influenced by the child's severity of illness at the time of the consent decision. DESIGN AND SETTING: Observational study in a multidisciplinary tertiary pediatric and neonatal intensive care. PATIENTS AND PARTICIPANTS: Parents of 421 children (age range from preterm to 18 years) were asked to consent for participation in a study focusing on measuring their child's nutritional status within 24 h after admission to the ICU. Over 20% of the parents (n=88) refused consent, most of them because they expected the study to be too burdensome for their child. MEASUREMENTS AND RESULTS: Patient and disease characteristics were comparable in the children for whom consent had or had not been obtained. A higher illness severity score did not decrease the probability of obtaining informed consent, but parents of children with a history of disease were 3.2 times less likely to consent. CONCLUSIONS: Parents of children with higher illness severity scores are not more likely to decline permission to include their child in clinical observational research on the ICU. History of disease and subjectively perceived burden to the child are important factors that must be considered. PMID- 15891863 TI - [Malignant fibrous histiocytoma in the parotid gland. Case series and literature review]. AB - BACKGROUND: Malignant fibrous histiocytoma (MFH), a soft tissue sarcoma that is predominantly localized in the extremities and retroperitoneum, rarely occurs in the head and neck. This study presents the clinicopathological features of three patients with MFH of the parotid gland treated at the ENT department of the University Hospital of Muenster (Germany) between 1991 and 2002. PATIENTS: The clinical course of all three patients was defined by a rapidly growing mass in the parotid area. Surgical therapy was the first treatment of choice. In two patients, radical parotidectomy was performed, whereas one patient underwent partial parotidectomy. Selective neck dissection was performed in one case. In two cases, post-surgical treatment involved radiation and/or chemotherapy. Two patients died as a consequence of local recurrence within the first year after diagnosis, whereas one patient is alive and free of disease after a follow up of 14 months. CONCLUSION: Our own experiences, taken together with those reported in the literature, suggest that clear surgical margins are probably the most important factor for avoiding a recurrence and to improve disease free survival. PMID- 15891864 TI - [Value of sentinel node biopsy for urological tumors]. AB - Gamma probe-guided lymphadenectomy of prostate cancer that is presumed to be localized furnishes evidence that lymphogenous spread of the disease is present considerably more often and earlier than previously assumed, even when the clinical stage is considered localized. Multiinstitutional trials have confirmed that in principle sentinel lymphadenectomy on its own is able to detect lymph node positive patients with minimal complications and a sufficient degree of certainty. Sentinel lymphadenectomy for penile cancer is an undemanding surgical procedure and in contrast to inguinal lymphadenectomy can be considered minimally invasive. Decisions on indication and necessity for an additional inguinal lymphadenectomy depending on tumor stage and local findings in the inguinal lymph nodes are handled quite differently in various centers and should be further standardized. The most recent studies on sentinel lymphadenectomy for urinary bladder and testicular cancer demonstrate that on principle the procedure is likely feasible also for these tumor entities. Whether it is possible to replace standard treatment methods with these procedures or at least have them serve an ancillary function remains to be determined in further investigations. Basically, the premise holds true that for all urological tumor entities before standard diagnostic techniques are abandoned, the value of exclusively performing sentinel lymphadenectomy must be adequately validated. It does not suffice to rely on the results from other working groups. It is in fact essential that the accuracy of the method - the feasibility of which can be influenced by numerous factors - be assessed by comparison with a standard lymphadenectomy performed in one's own center. PMID- 15891865 TI - [Lymphadenectomy for penile cancer. Diagnostic and prognostic significance as well as therapeutic benefit]. AB - Lymphadenectomy is an essential part of diagnosis and treatment of the squamous cell carcinoma of the penis. Lymphadenectomy is performed depending on various characteristics of penile cancer such as depth of invasion, tumor grade, invasion into the corpora cavernosa, invasion into vascular and lymphatic vessels. In case the inguinal lymphnodes are not palpable a modified lymphadenectomy is indicated. The limits of lymphadenectomy are extended to the radical type of dissection when the frozen section indicates cancer. Inguinal lymphadenectomy is always performed on both sides. Are more than 2 nodes positive the lymphnodes in the true pelvis have to be resected as well. The dynamic sentinel lymphnode dissection may replace the modified approach in case randomized prospective studies will confirm the initial positive results and morbidity can be reduced as well. The immediate lymphadenectomy is superior to the delayed lymphadenectomy (palpable nodes during followup) in terms of local recurrence and survival. According to the risk profile patients with palpable inguinal lymphnodes can be initially managed conservatively. In case the lymphnodes remain palpable, lymphadenectomy is indicated. In this situation it is reasonable to perform imaging studies of the pelvis and abdomen for adequate planning of the surgical approach. Neoadjuvant chemotherapy is reasonable for patients with bulky nodes fixed to the skin or fascia because this improves respectability, freedom from local recurrence and increases survival. Adjuvant chemo- and/or radio-therapy are reserved for extended disease or palliative situations. PMID- 15891866 TI - [Value of sentinel node biopsy considering melanoma management as an example]. AB - Sentinel lymph node biopsy has replaced elective lymph node dissection in the management of melanoma. It provides an exact staging method of the regional lymph basin and is associated with low morbidity. Patients who are found to have a positive sentinel lymph node can selectively undergo regional lymph node dissection. Sentinel lymph node biopsy also provides the means to stratify patients into homogeneous groups for adjuvant therapies. Sentinel lymph node involvement is regarded as an independent prognostic factor. The therapeutic value of subsequent surgical and medical therapies in patients with positive sentinel nodes is currently being investigated in clinical trials. PMID- 15891867 TI - [The follow-up of PSA tests for the early detection of prostate cancer. The responsibility of the physician to explain treatment methods which are not covered by public health insurance plans]. PMID- 15891868 TI - Determination of copper in powdered chocolate samples by slurry-sampling flame atomic-absorption spectrometry. AB - Chocolate is a complex sample with a high content of organic compounds and its analysis generally involves digestion procedures that might include the risk of losses and/or contamination. The determination of copper in chocolate is important because copper compounds are extensively used as fungicides in the farming of cocoa. In this paper, a slurry-sampling flame atomic-absorption spectrometric method is proposed for determination of copper in powdered chocolate samples. Optimization was carried out using univariate methodology involving the variables nature and concentration of the acid solution for slurry preparation, sonication time, and sample mass. The recommended conditions include a sample mass of 0.2 g, 2.0 mol L(-1) hydrochloric acid solution, and a sonication time of 15 min. The calibration curve was prepared using aqueous copper standards in 2.0 mol L(-1) hydrochloric acid. This method allowed determination of copper in chocolate with a detection limit of 0.4 microg g(-1) and precision, expressed as relative standard deviation (RSD), of 2.5% (n = 10) for a copper content of approximately 30 microg g(-1), using a chocolate mass of 0.2 g. The accuracy was confirmed by analyzing the certified reference materials NIST SRM 1568a rice flour and NIES CRM 10-b rice flour. The proposed method was used for determination of copper in three powdered chocolate samples, the copper content of which varied between 26.6 and 31.5 microg g(-1). The results showed no significant differences with those obtained after complete digestion, using a t test for comparison. PMID- 15891869 TI - Development and validation of a selective and robust LC-MS/MS method for quantifying amlodipine in human plasma. AB - A liquid chromatographic-tandem mass spectrometric method (LC-MS/MS) for quantifying amlodipine in human plasma was developed and validated. Sample preparation was based on liquid-liquid extraction using NaOH and a mixture of ethyl acetate/hexane (80/20; v/v). Chromatography was performed on a C-18 analytical column and the retention times were 1.9 and 3.0 min for amlodipine and nimodipine (internal standard), respectively. The ionization was optimized using ESI(+) and enhanced selectivity was achieved using tandem mass spectrometric analysis via two MRM functions, 409 --> 238 and 418 --> 343 for amlodipine and nimodipine. The calibration curve ranged from 0.2 to 20.0 ng/mL. The inter-day precision and accuracy and the relative standard deviation (RSD) were <15%. The analyte was shown to be stable over the time-scale of the whole procedure. The robustness of the method was demonstrated by the good reproducibility of the results obtained during the analysis of clinical samples. PMID- 15891870 TI - Amperometric detection in TMB/HRP-based assays. AB - 3,3',5,5'-Tetramethylbenzidine (TMB) is the most commonly used chromogen for horseradish peroxidase (HRP) and so its performance as an electrochemical substrate was evaluated. Measurements of HRP activity in solution were carried out by using an amperometric detector coupled to a flow injection analysis (FIA) system. The enzymatic product was easily detected at a potential of +0.1 V (vs. Ag-pseudoreference electrode) at a bare screen-printed electrode placed in a homemade electrochemical flow cell. A high flow rate (4.3 mL min(-1)) of 0.5 M H2SO4 was used to obtain repeatable signals and a short analysis time. The detection limit achieved after 15 min of incubation was 2x10(-14) M of HRP. The applicability of the amperometric detector to ELISAs was demonstrated by using a commercially available kit for the quantification of interleukin-6 (IL-6) without modifying the kit manufacturer's protocol or the reagents for this test. PMID- 15891871 TI - Neural representation of response category and motor parameters in monkey prefrontal cortex. AB - Conditional motor behavior, in which the relationship between stimuli and responses changes arbitrarily, is an important component of cognitive motor function in primates. It is still unclear how cognitive processing for conditional motor control determines movement parameters to directly specify motor output. To address this issue, we studied the neuronal representation of motor variables relating to conditional motor control and also directly to the metrics of motor output in prefrontal cortex (PFC). Monkeys were required to generate a force that fell within one of two categories ("small" and "large"). We found that most PFC neurons were activated as a function of force category, suggesting a role in conditional motor control. At the same time, we found that activity in many PFC neurons varied continuously with the force that was eventually produced, suggesting they participated in specifying the metrics of movements as they were executed. The results suggest that the PFC neural population encodes both "what" motor response should be performed and "how" the selected movement should be realized immediately after the visual instruction. PMID- 15891872 TI - The use of non-motion-based cues to pre-programme the timing of predictive velocity reversal in human smooth pursuit. AB - Human smooth pursuit eye movements are principally driven by visual feedback and cannot normally be initiated at will. However, when tracking periodic motion, smooth eye movements reverse direction prior to target reversal, driven by anticipation, not visual feedback. Here, we investigate cognitive control over such eye reversals. Target stimuli were discrete double ramps-constant speed (30 degrees /s) rightwards followed by similar leftward movement, reversal time ranging from 420 ms to 840 ms. Three experimental conditions were examined. In the precued condition, double ramps of randomised reversal time were presented. Prior to ramp presentation audio precues were given with an interval indicating start and reversal time of the unseen, upcoming double ramp. Subjects were able to use these cues to voluntarily control timing of anticipatory eye reversal, so that when occasional false precues gave underestimates of target reversal time, eye reversal occurred before target reversal. Precued eye reversal times were comparable to those in a second, predictable condition, in which double ramps with identical reversal time were given repeatedly without precues. In contrast, reversal occurred much later in a third, reactive condition, also without precues, when unexpected early target reversals occurred sporadically within a series having identical, predictable reversal times. The findings provide evidence that timing of anticipatory smooth eye movement, both at the start of the double-ramp and at its reversal, can be independently controlled at will using non-motion-based timing cues. PMID- 15891873 TI - Disruptions in joint control during drawing arm movements in Parkinson's disease. AB - Impairments in control of multi-joint arm movements in Parkinson's Disease (PD) were investigated. The PD patients and age-matched elderly participants performed cyclical arm movements, tracking templates of a large circle and four differentially oriented ovals on a horizontal table. The wrist was immobilized and the movements were performed with shoulder and elbow rotations. The task was performed with and without vision at a cycling frequency of 1.5 Hz. Traces of the arm endpoint, joint-motion parameters represented by range of motion and relative phase, and joint-control characteristics represented by amplitude and timing of muscle torque were analyzed. The PD patients provided deformations of the template shapes that were not observed in movements of elderly controls. The deformations were consistent for each shape but differed across the shapes, making quantification of impairments in the endpoint movement difficult. In contrast, the characteristics of joint control and motion demonstrated systematic changes across all shapes in movements of PD patients, although some of these changes were observed only without vision. A specification of the PD influence was observed at the level of joint control and it was not distinguishable in joint and endpoint motion, because of the property of multi-joint movements during which control at each joint influences motion at the other joints. The results suggest that inability of PD patients to provide fine muscle torque regulation coordinated across the joints contributes to the altered endpoint trajectories during multi-joint movements. The study emphasizes the importance of the torque analysis when deficits in multi-joint movements are investigated, because specific impairments that can be detected in joint-control characteristics are difficult to trace in characteristics of joint and endpoint kinematics, because of interactions between joint motions. PMID- 15891874 TI - The questionable use of antihypertensive drugs in the Italian Liguria Region in the year 2003. PMID- 15891875 TI - Relationship between calcineurin inhibition and plasma endothelin concentrations in cyclosporine-A-treated kidney transplant patients--a comment. PMID- 15891876 TI - Diffuse vertebral body edema due to calcified intraspongious disk herniation. AB - We describe the case of a patient with a recent history of high back pain, with magnetic resonance imaging (MRI) of the thoracic spine showing intervertebral disk herniation into the spongious bone of the vertebral body of T9 that might have caused diffuse, low signal intensity on fluid-attenuated inversion recovery T1-weighted (FLAIR-T1W) images, high signal intensity magnetic resonance (MR) on T2-weighted (T2W) images and T2-weighted fat-suppressed images (T2W-FSIs) and marked enhancement on the vertebral body of T9 with gadolinium on T1-weighted fat suppressed images (T1W-FSIs) images. Those findings suggested diffuse edema and might be indistinguishable from tumoral or inflammatory diseases, but the plain films and the reformatted sagittal computed tomography scans of the thoracic spine were helpful to show a calcified part of the intervertebral disk migrating into the vertebral body of T9. The patient made full recovery from the symptoms after conservative treatment and at the follow-up MRI showed normalization of the bone marrow signal intensity of the vertebral body of T9. PMID- 15891877 TI - Comparing axillary and mediastinal lymphadenopathy on CT in children with suspected pulmonary tuberculosis. AB - BACKGROUND: Radiographic demonstration of mediastinal lymphadenopathy is important for the diagnosis of pulmonary tuberculosis (PTB). Plain radiographs are unreliable for this and CT, which is relatively more expensive and carries a high radiation burden, remains the gold standard. No studies correlating the presence of axillary with mediastinal lymphadenopathy have been reported. Such a correlation would allow for clinical or ultrasound diagnosis of PTB via the axilla. OBJECTIVE: To correlate the presence of axillary lymphadenopathy with mediastinal lymphadenopathy in children with suspected PTB. MATERIALS AND METHODS: CT scans were performed and reviewed in 100 children (prospectively recruited) with suspected PTB. The axilla and mediastinum were reviewed separately by covering the non-relevant sections on the CT scans prior to reading. Only nodes greater than 1 cm were regarded as pathological. RESULTS: Mediastinal lymphadenopathy was present in 46% of children; 70% had lymphadenopathy in either axilla. Bilateral axillary lymphadenopathy was identified in 47%. Axillary lymphadenopathy showed a sensitivity of 74% and a specificity of 33% for the presence of mediastinal adenopathy. Bilateral axillary adenopathy had a sensitivity of 50% and a specificity of 56%. CONCLUSIONS: Axillary lymphadenopathy has a moderate sensitivity and low specificity for the presence of mediastinal and hilar lymphadenopathy in children with suspected PTB. Further research should be aimed at correlating ultrasound-detected axillary lymphadenopathy with FNA results in children. PMID- 15891878 TI - Lumbar and iliac artery aneurysms in Menkes' disease: endovascular cover stent treatment of the lumbar artery aneurysm. AB - We report lumbar and iliac artery aneurysms in a 3-month-old boy with Menkes' disease. The iliac artery aneurysm thrombosed spontaneously, documented by follow up colour Doppler sonography. The lumbar artery aneurysm was successfully treated using a cover stent. There was no filling of the lumbar artery aneurysm and no stenosis of the cover stent during the 9-month follow-up. PMID- 15891879 TI - Sirenomelia (symelia dipus). PMID- 15891880 TI - Adoptive T cell therapy of solid cancers. AB - The development of immune-based approaches for the treatment of cancer has been actively investigated for many years. One strategy that has emerged as a potentially effective strategy for the treatment of aggressive established malignancies is adoptive T cell therapy. The power of this approach has been repeatedly observed in preclinical animal models. However, moving from homogeneous animal models to the heterogeneous human clinical setting has been very difficult. It is only in recent times that we have been able to pinpoint the problems of the clinical translation of adoptive T cell therapy. Some of the major problems are sources of tumor-specific T cells, ex vivo expansion, persistence, and anti-tumor activity. This review overviews the nature of these problems and some of the emerging solutions. PMID- 15891881 TI - Virulizin, a novel immunotherapy agent, activates NK cells through induction of IL-12 expression in macrophages. AB - Virulizin, a novel biological response modifier, has demonstrated significant antitumor efficacy in a variety of human tumor xenograft models including melanoma, pancreatic cancer, breast cancer, ovarian cancer and prostate cancer. The significant role of macrophages and NK (Natural killer) cells was implicated in the antitumor mechanism of Virulizin where expansion as well as increased activity of macrophages and NK cells were observed in mice treated with Virulizin. Depletion of macrophages compromised Virulizin-induced NK1.1+ cell infiltration into xenografted tumors and was accompanied by reduced antitumor efficacy. In the present study, involvement of macrophages in NK cell activation was investigated further. We found that depletion of NK cells in CD-1 nude mice by anti-ASGM1 antibody significantly compromised the antitumor activity of Virulizin. Cytotoxicity of NK cells isolated from Virulizin-treated mice was enhanced against NK-sensitive YAC-1 cells and C8161 human melanoma cells, but not against NK-insensitive P815 cells. An increased level of IL-12beta was observed in the serum of mice treated with Virulizin. IL-12 mRNA and protein levels were also increased in peritoneal macrophages isolated from Virulizin-treated mice. Moreover, Virulizin-induced cytotoxic activity of NK cells isolated from the spleen was abolished when an IL-12 neutralizing antibody was co-administered. In addition, depletion of macrophages in mice significantly impaired Virulizin induced NK cell cytotoxicty. Taken together, the results suggest that Virulizin induces macrophage IL-12 production, which in turn stimulates NK cell-mediated antitumor activity. PMID- 15891882 TI - Release of iC3b from apoptotic tumor cells induces tolerance by binding to immature dendritic cells in vitro and in vivo. AB - Chemo- as well as immunotherapeutical approaches induce apoptosis in tumor cells. Apoptotic cells are known to activate homologous complement and to be opsonized with iC3b. Since maturation of dendritic cells (DC) can be inhibited by binding of iC3b to the complement receptor 3 (CR3, CD11b/CD18) and because immature DC induce tolerance, we investigated the induction of tolerance after pulsing DC with apoptotic cells in the presence or absence of native serum. Apoptosis in pancreatic carcinoma cells was induced either by heat-stress, chemotherapy or anti-Her2 antibody. Monocyte-derived DC were pulsed with apoptotic cells with or without native serum. DC were analyzed for the maturation state by flow cytometry and the cytotoxic activity was determined. Tolerance was prevented by addition of substances such as anti CD11b or N-acetyl-D-Glucosamine (NADG) which block iC3b binding to CR3. Furthermore, binding of iC3b from apoptotic cells to DC was blocked in a syngeneic pancreatic carcinoma mouse model. All of the former strategies for apoptosis induction resulted in iC3b release. Pulsing DC with apoptotic cells in the presence of serum prevents maturation of DC and induces finally tolerance. This tolerance could be prevented almost completely by blocking the interaction of iC3b with the CR3 receptor. This could be shown as well in an immunocompetent mouse model. Chemo- as well as immunotherapeutical approaches induce apoptosis in tumor cells. Release of iC3b from apoptotic tumor cells prevents fully maturation of DC and immature DC induce antigen-specific silencing or tolerance. Blocking of iC3b-binding could mostly prevent this effect. PMID- 15891883 TI - Allogeneic gastric cancer cell-dendritic cell hybrids induce tumor antigen (carcinoembryonic antigen) specific CD8(+) T cells. AB - The development of protocols for the ex vivo generation of dendritic cells (DCs) has led to intensive research of their potential use in immunotherapy. Accumulating results show the efficacy of this treatment on melanomas which are highly immunogenic. However, its efficacy remains unclear in other tumors. In this study, allogeneic gastric cancer cell-DC hybrids were used to determine the efficacy of this type of immunotherapy in gastric cancer. Fusion cells of DC and allogeneic gastric cancer cells were generated by polyethylene glycol (PEG) and electrofusion. These hybrids were used to induce tumor associated antigen (TAA) specific cytotoxic T lymphocytes (CTLs). The DCs were successfully fused with the allogeneic gastric cancer cells resulting in hybrid cells. These hybrid cells were functional as antigen-presenting cell because they induced allogeneic CD4(+) T cells proliferation. CD8(+) T cells stimulated by the MKN-45-DC hybrid cells were able to kill MKN-45 when used for immunization. The CTLs killed another gastric cancer cell line, MKN-1, as well as a melanoma cell line, 888mel, suggesting the recognition of a shared tumor antigen. MKN-45 specific CTLs can recognize carcinoembryonic antigen (CEA), indicating that the killing is due to tumor antigens as well as alloantigens. This approach suggests the possible use of allogeneic gastric cancer cell-DC hybrids in DC based immunotherapy for gastric cancer treatment. PMID- 15891884 TI - Development of a whole cell vaccine for acute myeloid leukaemia. AB - We describe the modification of tumour cells to enhance their capacity to act as antigen presenting cells with particular focus on the use of costimulatory molecules to do so. We have been involved in the genetic modification of tumour cells to prepare a whole cell vaccine for nearly a decade and we have a particular interest in acute myeloid leukaemia (AML). AML is an aggressive and difficult to treat disease, especially, for patients for whom haematopoietic stem cell (HSC) transplant is not an option. AML patients who have a suitable donor and meet HSC transplant fitness requirements, have a 5-year survival of 50%; however, for patients with no suitable donor or for who age is a factor, the prognosis is much worse. It is particularly poor prognosis patients, who are not eligible for HSC transplant, who are likely to benefit most from immunotherapy. It would be hoped that immunotherapy would be used to clear residual tumour cells in these patients in the first remission following standard chemotherapy treatments and this will extend the remission and reduce the risk of a second relapse associated with disease progression and poor mortality rates. In this symposia report, we will focus on whole cell vaccines as an immunotherapeutic option with particular reference to their use in the treatment of AML. We will aim to provide a brief overview of the latest data from our group and considerations for the use of this treatment modality in clinical trials for AML. PMID- 15891885 TI - CD3 x CD28 cross-interacting bispecific antibodies improve tumor cell dependent T cell activation. AB - Bispecific antibodies (Bs-Abs) containing an anti-CD3 and an anti-TAA specificity can recruit T cells to the tumor for cancer immunotherapy. To be effective, efficient activation at the tumor site is a prerequisite. This can be achieved by triggering both the T-cell receptor and the co-stimulatory molecule CD28. We engineered two recombinant cross-interacting Bs-Abs (CriBs-Abs) by incorporating a peptide tag and its cognate single-chain variable fragment (scFv), respectively, into a pair of (tumor x CD3) and (tumor x CD28) binding Bs-Abs. A 30-fold lower concentration of the activating CriBs-Ab as compared to non interacting Bs-Ab was sufficient for strong T-cell activation in the presence of tumor cells. One thousand-fold higher concentrations of both CriBs-Abs were required for marginal T-cell activation (70-fold below maximal response) in the absence of tumor cells. An optimized stoichiometry (1 : 1000) of activating versus co-stimulating CriBs-Ab thus allowed low doses of activating CriBs-Ab to induce tumor-cell dependent T-cell activation when used in combination with high concentrations of the pre-targeted co-stimulating CriBs-Ab in vitro. This indicates a large window of operation in which only tumor cell dependent T-cell activation is induced and systemic tumor cell independent T-cell activation is avoided, while ensuring optimal activation with a low concentration of the activating CriBs-Ab, which has the highest potential to induce toxic effects in vivo. PMID- 15891886 TI - Cyclooxygenase inhibitors modulate NK activities that control metastatic disease. AB - Cyclooxygenase (COX) inhibitors have demonstrated efficacy in models of human cancer but the relevant mechanisms have not all been elucidated. Both Cox dependent as well as Cox-independent mechanisms have been implicated. Using a syngeneic model of metastatic breast cancer, we have investigated the effect of Cox inhibitors on NK functions that are critical to the control of metastatic disease. NK recognition of target cells is governed by a balance of activating and inhibiting receptors that bind ligands including MHC class I. We now show that treatment of tumor cells with the nonselective COX-1/COX-2 inhibitor indomethacin or the selective COX-2 inhibitor celecoxib leads to decreased expression of the MHC class I molecules Ld and Kd . Downregulated class I expression is associated with concomitant increased sensitivity to NK cell mediated lysis. Both COX inhibitors limit tumor metastasis and this therapeutic effect is dependent on NK but not T cell function. Antimetastatic activity is also lost in the absence of interferon- gamma (IFN-gamma). Both COX inhibitors also suppress local tumor growth of subcutaneously implanted mammary tumor cells in immune competent Balb/cByJ mice. This therapeutic activity is lost in the absence of either CD4+ or CD8+ T cells, but is not compromised by the loss of NK activity. Thus, the mechanism of tumor inhibition differs in the context of local versus metastatic disease. Taken together, these findings are consistent with a mechanism not previously described, whereby COX inhibitors may relieve MHC mediated inhibition of NK cytotoxicity leading to recognition and lysis of metastatic tumor cells. PMID- 15891887 TI - Prognosis of patients with del(5q) MDS and complex karyotype and the possible role of lenalidomide in this patient subgroup. AB - The survival of patients with myelodysplastic syndromes is strongly affected by chromosomal abnormalities. Patients with an isolated del(5q31) have a favourable prognosis that worsens with the addition of another chromosomal abnormality. It has been reported that both patients with isolated del(5q31) and those with one single additional chromosomal abnormality achieve hematological and cytogenetic remissions with lenalidomide therapy. Whether this translates into improved overall survival of the patient population is unclear. We analysed data of 25 patients with myelodysplastic syndrome and complex chromosomal abnormalities including del(5q31) and show that their median survival is between 7 and 8 months, irrespective of the medullary blast count. Furthermore, we present data of a patient with complex karyotypic anomalies inclusive of del(5q31) treated with lenalidomide who achieved complete cytogenetic remission. This cytogenetic remission was diagnosed after 6 months, and the hematological response is ongoing at 9 months of therapy at a dose of 5 mg p.o. daily. We conclude that lenalidomide has the potential to induce sustained hematological and cytogenetic remissions in the poor prognosis MDS subgroup of del(5q31) patients with complex chromosomal anomalies and that this is likely to improve overall survival. PMID- 15891888 TI - MRI for short-term follow-up of acute pulmonary embolism. Assessment of thrombus appearance and pulmonary perfusion: a feasibility study. AB - Tha aim of this study was to demonstrate the feasibility of MRI for short-term follow-up examinations in patients with acute pulmonary embolism (PE), and to assess temporal changes of pulmonary perfusion and thrombus characteristics that may be helpful in determining thrombus age. Thirty-three patients (15 female, 18 male, mean age 59.4 years) with acute PE were examined initially and 1 week later using both 16-row computed tomography (CT) and MRI with magnetic resonance angiography (MRA), real-time MRI and magnetic resonance (MR) pulmonary perfusion imaging. MRA and MR pulmonary perfusion used contrast-enhanced 3D flash sequences, and real-time MRI used true fast imaging with steady-state precession sequences (repetition time/echo time 3.1/1.5, bandwidth 975 Hz, 256 matrix size, acquisition time 0.4 s per image) in three orthogonal planes. Follow-up examinations were feasible for all patients. Diagnosis of PE was concordant between MRI and CT in all patients. The signal intensity of embolic material increased after 1 week for real-time MRI [132+/-5 vs. 232+/-22 (standard error of the mean), p<0.001], but not significantly for MRA. MR pulmonary perfusion of areas affected by PE increased (area under the curve initially 9.6+/-7.4, at follow-up 40.7+/-7.6, p<0.001). A decreasing time-to-peak in normal lung areas (15.7+/-0.96 and 13.2+/-0.55, respectively, p<0.05) indicated systemic circulatory effects of PE, and subsiding pulmonary artery obstruction improved arterial inflow for the entire lung. Follow-up examinations of patients with acute PE are feasible with MRI, and a relation between thrombus appearance and thrombus age can be implied. PMID- 15891889 TI - Changes in caridac output and hemolymph flow during hypoxic exposure in the gravid grass shrimp, Palaemonetes pugio. AB - The cardiovascular response of decapod crustaceans to hypoxic exposure is well documented; however, information is limited concerning the influence of reproductive state on cardiovascular demands during hypoxic exposure. Given the additional metabolic demand of reproduction, we investigated the cardiovascular adjustments employed by gravid grass shrimp Palaemonetes pugio to maintain oxygen delivery during hypoxic stress. Cardiac output values were elevated in gravid compared to nongravid grass shrimp. Gravid grass shrimp were exposed to hypoxia and the stroke volume, heart rate, cardiac output and hemolymph flow were determined using video-microscopy and dimensional analysis. Oxygen consumption rates were determined using respirometry. There where no changes in the cardiac output values of gravid females until reaching 6.8 kPa O2, with a significant redistribution of hemolymph flow at 13.7 kPa O2. Flow was significantly decreased to the anterior lateral arteries that supply the ovaries and hepatopancreas, the anterior aorta and the posterior aorta. The redistribution of hemolymph flow away from these vessels results in an enhanced hemolymph flow to the sternal artery that supplies the ventral segmental system, the gills, the buccal apparatus and the ventral nerve cord. The data suggest that during hypoxic stress, gravid females place a priority on survival. PMID- 15891890 TI - Penetrating pharyngeal injuries in children: trivial trauma leading to devastating complications. AB - Pharyngeal perforations are uncommon in children and are usually secondary to instrumentation or external penetrating injuries. A delay in management can lead to life-threatening complications such as retropharyngeal abscess, mediastinitis, and airway compromise. We report three children who had pharyngeal perforation due to apparently innocuous injury and who developed serious complications. A high index of suspicion for a significant pharyngeal injury, use of lateral soft tissue x-ray films of the neck, and flexible endoscopy in the emergency room can assist in early diagnosis when evaluating patients with oropharyngeal and penetrating neck injury. Prompt administration of broad-spectrum antibiotics, local drainage, and debridement can avert life-threatening complications. PMID- 15891891 TI - Epidural emphysema associated with subcutaneous emphysema following foreign body in the airway. AB - Epidural emphysema is a rare condition and usually accompanies pneumothorax, subcutaneous emphysema (SCE), or pneumomediastinum. We report a child who presented with epidural emphysema (pneumorachis) and SCE. The child was subsequently found to have a foreign body obstructing the airway, which was the likely cause of the pneumorachis. Thus, foreign body aspiration should be considered in an infant with SCE, and early intervention in the form of bronchoscopy should to be done to arrest the condition's progressive nature. PMID- 15891892 TI - Epidermal growth factor and bombesin act synergistically to support intestinal adaptation in rats with massive small bowel resection. AB - Intestinal adaptation is the most important event in short bowel syndrome following a massive small bowel resection. Effects of various growth factors and their synergism have been well documented in intestinal adaptation. This study aimed to compare the effect of two different trophic agents, epidermal growth factor (EGF) and bombesin (BBS), on intestinal adaptation following massive intestinal resection. Sprague-Dawley male rats were assigned to one of four groups after a 75% small bowel resection. Either EGF (90 microg/kg), BBS (10 microg/kg), EGF+BBS, or bovine serum albumin (BSA) were injected subcutaneously three times a day. The animals were killed 10 days after the operation. Weight loss and morphologic parameters such as mucosal thickness, villus height, crypt depth, villus-to-crypt ratio, and muscularis propria height were measured. In the EGF+BBS group, mucosal thickness was found to be significantly increased compared with the other study groups (p<0.05). Similarly, villus height was significantly increased only in the EGF+BBS group (p<0.05). In the BBS group, both villus height and mucosal thickness showed a slight increase, but the values were not statistically significant compared with the vehicle-treated group. There were no significant differences in any of the remaining parameters between the groups. The results of this study indicate that the gut hormones EGF and BBS act synergistically in facilitating the adaptive response of the remnant ileum to massive intestinal resection. PMID- 15891893 TI - Prognostic factors of liver metastases from uveal melanoma. AB - OBJECTIVES: This study was designed to assess survival and identify prognostic factors for liver metastases diagnosed by systematic screening in uveal melanoma patients. METHODS: Among 602 consecutive patients treated over 10 years for uveal melanoma and followed by systematic semi-annual hepatic screening (abdominal ultrasonography), 63 (10.5%) developed liver metastases; these patients form the basis of this study. Factors including patient demographics, characteristics of the uveal tumor, metastasis-free interval, severity of liver metastatic involvement, and treatments of metastases were studied retrospectively regarding their prognostic value, using univariate (Kaplan-Meier method) and multivariate (Cox model) analyses. RESULTS: Thirty-five patients (55.6% of the metastatic population) received systemic chemotherapy or best supportive care only; 14 patients (22.2% of the metastatic population) diagnosed with diffuse liver involvement had cytoreductive surgery and intra-arterial chemotherapy; 14 (22.2% of the metastatic population) had complete surgical removal of liver metastases followed by postoperative intra-arterial chemotherapy. No significant surgical complications were experienced. The median overall survival after diagnosis of liver metastases was 15 months. It reached 25 months for selected patients with complete resection (P=0.0002). In this cohort of 63 patients, ten or fewer preoperatively diagnosed metastases and primary uveal melanoma not involving the ciliary body were independently associated with better prognosis. CONCLUSIONS: This study suggests that selected patients with screened liver metastases from uveal melanoma may benefit from aggressive treatment, including surgery. The two independent favorable prognostic factors are fewer than ten metastases at screening and the absence of ciliary body involvement. PMID- 15891894 TI - Quantification and anatomic distribution of choroidal abnormalities in patients with type I neurofibromatosis. AB - BACKGROUND: Choroidal abnormality manifesting as a bright patchy lesion under infrared monochromatic light has previously been described in neurofibromatosis type I patients in whom the choroid appears normal under conventional ophthalmoscopic examination or on the fluorescein angiogram. We investigated the correlation between patient age and the number of choroidal abnormalities, as well as the anatomic distribution of choroidal abnormalities in the fundus. METHODS: We examined the fundus of 28 eyes in 14 patients with neurofibromatosis type I. Patients ranged in age from 2 to 38 years and were examined between April 2001 and April 2002 by confocal scanning laser ophthalmoscopy with infrared monochromatic light (780 nm wavelength). We divided the fundus into five regions (one within the retinal vascular arcade and those supero-temporal, infero temporal, supero-nasal, and infero-nasal to it), and lesions on the border between regions were assigned to the region containing the greater part of the lesion. We studied the total number of choroidal abnormalities and the correlation between the total number and age. RESULTS: A positive correlation was found between the total number of choroidal abnormalities and age (Spearman rank correlation coefficient, r=0.6209, P=0.0178). There was a significantly greater number of choroidal abnormalities in the arcade region than in the other four regions (ANOVA, P<0.001). CONCLUSIONS: Choroidal abnormalities tend to increase with age and are most often observed within the vascular arcade. PMID- 15891895 TI - Human sterol 12a-hydroxylase (CYP8B1) is mainly expressed in hepatocytes in a homogenous pattern. AB - The liver is the only organ where the complete synthesis of bile acids takes place. The present study was undertaken to investigate whether regional differences exist within the individual human hepatic lobuli regarding the pattern of expression of sterol 12alpha-hydroxylase (CYP8B1), a key enzyme in bile acid synthesis. A specific anti-human CYP8B1 peptide antiserum was developed and used for Western blotting and hepatic immunostaining of livers from various patients. CYP8B1 in human liver was expressed in the cytoplasm of hepatocytes with an even nonzonal distribution within the liver lobulus. Pericentral expression was confirmed for CYP2E1. A weak staining was noted in cholangiocytes and Kupffer cells. Previous studies on hepatic CYP27A1 and CYP7A1 in rats have shown a zonal expression, primarily in the pericentral region. Our studies indicate a different pattern for CYP8B1 expression in human liver, which was even rather than zonal. PMID- 15891896 TI - Microsomal triglyceride transfer protein expression in mouse intestine. AB - Immunohistochemical and biochemical approaches were utilized to compare the expression of microsomal triglyceride transfer protein (MTP) and cellular retinol binding protein II (CRBPII) with the expression of apolipoprotein (apo)B and apoA I along the entire length of the small intestine in mice. MTP is expressed in villus-associated enterocytes along the length of the small intestine. Maximal expression occurs within the first 20% of the intestine and decreases to less than 3% of maximum in the distal third of the intestine. The expression of CRBPII is nearly identical with that of MTP. Peak expression of apoB and apoA-I occurs in the first 25% of the intestine; however, expression in the most distal segments of the intestine is 10%-15% of maximum expression. In mice fed a Western diet for 3 weeks the expression of MTP and CRBPII was elevated in the distal regions of the intestine, whereas the expression patterns for apoB and apoA-I were similar to those found in mice on control diets. We conclude that the patterns of expression, as well as the regulation of MTP and CRBPII, are similar. However, the expression and regulation of these two proteins differ from those of apoB and apoA-I. In particular, the expression of MTP is not coordinated with the expression of apoB, even though the two proteins are essential for the assembly and secretion of chylomicrons. PMID- 15891897 TI - Identification of three shikimate kinase genes in rice: characterization of their differential expression during panicle development and of the enzymatic activities of the encoded proteins. AB - The shikimate pathway is common to the biosynthesis of the three aromatic amino acids and that of various secondary metabolites in land plants. Shikimate kinase (SK; EC 2.7.1.71) catalyzes the phosphorylation of shikimate to yield shikimate 3 phosphate. In an attempt to elucidate the functional roles of enzymes that participate in the shikimate pathway in rice (Oryza sativa), we have now identified and characterized cDNAs corresponding to three SK genes--OsSK1, OsSK2, and OsSK3--in this monocotyledenous plant. These SK cDNAs encode proteins with different NH(2)-terminal regions and with putative mature regions that share sequence similarity with other plant and microbial SK proteins. An in vitro assay of protein import into intact chloroplasts isolated from pea (Pisum sativum) seedlings revealed that the full-length forms of the three rice SK proteins are translocated into chloroplasts and processed, consistent with the assumption that the different NH(2)-terminal sequences function as chloroplast transit peptides. The processed forms of all three rice proteins synthesized in vitro manifested SK catalytic activity. Northern blot analysis revealed that the expression of OsSK1 and OsSK2 was induced in rice calli by treatment with the elicitor N acetylchitoheptaose, and that expression of OsSK1 and OsSK3 was up-regulated specifically during the heading stage of panicle development. These results suggest that differential expression of the three rice SK genes and the accompanying changes in the production of shikimate 3-phosphate may contribute to the defense response and to panicle development in rice. PMID- 15891898 TI - Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica. AB - In order to characterize a specific extracellular 21-kDa calmodulin-binding protein (named: ECBP21) from Angelica dahurica L. suspension-cultured cells, the cDNA coding for the protein has been cloned. Here, Southern blot analysis shows that there are at least two copies of ECBP21 gene in Angelica genome. Using truncated versions of ECBP21 and synthetic peptide in CaM binding assays, we mapped the calmodulin-binding domain to a 16-amino acid stretch (residues 200 215) at the C-terminal region. The ECBP21 was localized in the cell wall area by the immunogold electron microscopy and by GFP labeling method. These results define ECBP21 as a kind of an extracellular calmodulin-binding protein (CaMBP). Furthermore, using Northern blot analysis, we examined the expression dynamics of ecbp21 during the incubation of Angelica suspension-cultured cells and the treatments with some growth regulators. The above studies further provide the molecular evidence for the existence of the gene coding for extracellular CaMBPs and imply a possible role for ECBP21. PMID- 15891899 TI - AtUTr2 is an Arabidopsis thaliana nucleotide sugar transporter located in the Golgi apparatus capable of transporting UDP-galactose. AB - The synthesis of noncellulosic polysaccharides and glycoproteins in the plant cell Golgi apparatus requires UDP-galactose as a substrate. We have cloned and characterized a nucleotide sugar transporter from Arabidopsis thaliana (L.) Heynh. named AtUTr2. Expression in tobacco and Saccharomyces cerevisiae and subsequent biochemical characterization indicate that AtUTr2 transports UDP galactose, but not UDP-glucose, UDP-N-acetyl glucosamine, UDP-xylose, UDP glucuronic acid, GDP-fucose or GDP-mannose. Experiments expressing an AtUTr2-GFP fusion protein in onion epidermal cells suggest that AtUTr2 is located in the Golgi apparatus. Finally, northern analysis indicates that the AtUTr2 transcript was more abundant in roots and calli although it was also present in other Arabidopsis organs but at lower levels. Therefore, AtUTr2 is a nucleotide sugar transporter capable of transporting UDP-galactose that may play an important role in the synthesis of galactose-containing glycoconjugates in Arabidopsis. PMID- 15891900 TI - The composition and timing of flower odour emission by wild Petunia axillaris coincide with the antennal perception and nocturnal activity of the pollinator Manduca sexta. AB - In the genus Petunia, distinct pollination syndromes may have evolved in association with bee-visitation (P. integrifolia spp.) or hawk moth-visitation (P. axillaris spp). We investigated the extent of congruence between floral fragrance and olfactory perception of the hawk moth Manduca sexta. Hawk moth pollinated P. axillaris releases high levels of several compounds compared to the bee-pollinated P. integrifolia that releases benzaldehyde almost exclusively. The three dominating compounds in P. axillaris were benzaldehyde, benzyl alcohol and methyl benzoate. In P. axillaris, benzenoids showed a circadian rhythm with an emission peak at night, which was absent from P. integrifolia. These characters were highly conserved among different P. axillaris subspecies and P. axillaris accessions, with some differences in fragrance composition. Electroantennogram (EAG) recordings using flower-blends of different wild Petunia species on female M. sexta antennae showed that P. axillaris odours elicited stronger responses than P. integrifolia odours. EAG responses were highest to the three dominating compounds in the P. axillaris flower odours. Further, EAG responses to odour samples collected from P. axillaris flowers confirmed that odours collected at night evoked stronger responses from M. sexta than odours collected during the day. These results show that timing of odour emissions by P. axillaris is in tune with nocturnal hawk moth activity and that flower-volatile composition is adapted to the antennal perception of these pollinators. PMID- 15891901 TI - Structural characterization, expression analysis and evolution of the red/far-red sensing photoreceptor gene, phytochrome C (PHYC), localized on the 'B' genome of hexaploid wheat (Triticum aestivum L.). AB - Phytochromes are a family of red/far-red light perceiving photoreceptors. The monocot phytochrome family is represented by three members, PHYA, PHYB and PHYC. We have isolated and characterized the first PHY gene member (TaPHYC) from common wheat, Triticum aestivum var. CPAN1676. It codes for a species of the photoreceptor, phyC, which is known to be light-stable in all plants analyzed so far. A sequence of 7.2 kb has been determined, which includes 3.42 kb of coding region. This is the second full-length PHYC gene sequenced from a monocot (first was from rice). TaPHYC gene shares structural similarities with the rice PHYC containing four exons and three introns in the coding region. The 5' UTR is 1.0 kb-long and harbors an upstream open reading frame (URF) encoding 28 aa. Southern blot analysis of TaPHYC indicates that it represents single locus in the wheat genome, although the possibility of additional loci cannot be completely ruled out. Chromosomal localization using nullisomic-tetrasomic lines of Triticum aestivum var. Chinese Spring places TaPHYC on chromosome 4B. PHYC represents a constitutively expressed gene in all the organs tested and under light/dark conditions. However, PHYC was found to be developmentally regulated showing maximal expression in 3-day-old dark-grown seedlings, which declined thereafter. In silico analysis has also been done to compare TaPHYC gene with the partial sequences known from other wheat species and cultivars. The presence of a topoisomerase gene immediately downstream of the PHYC gene, both in rice and wheat genomes, presents yet another example of synteny in cereals and its possible significance has been discussed. PMID- 15891902 TI - Global DNA methylation evaluation: potential complementary marker in differential diagnosis of thyroid neoplasia. AB - The implications of global DNA hypomethylation were recently reported in several models of tumorigenesis. Little is known about this epigenetic event in thyroid neoplasia. The study aimed to evaluate the status of global DNA methylation in several types of thyroid tumors using a monoclonal antibody specific for 5 methylcytidine (5-mc) and to define the diagnosis potential of this marker. 5-mc immunostaining scores were calculated in 17 papillary thyroid carcinomas (PTC), 6 follicular thyroid carcinomas (FTC), 16 follicular adenomas (FA), 19 nodular goiters (NG) and ten Hurthle cells adenomas (HCA). The expression of galectin-3 was also evaluated. Computerized image analysis showed a significant lower level of 5-mc immunostaining in thyroid carcinoma when compared with benign tumors or adjacent normal thyroid parenchyma (P<0.0001). Overall, 5-mc accuracy to distinguish malign from benign thyroid tumors was similar to that of galectin-3 (89% versus 87%, P>0.05). The combination of 5-mc with galectin-3 led to an excellent accuracy level of 96%. Among follicular neoplasia 5-mc accuracy to differentiate malign tumors trends to be higher than galectin-3 one (90% versus 66%, P=0.06). These data stress the necessity of epigenetic events evaluation among thyroid nodules and propose global DNA methylation assessment as a potential diagnostic tool to combine with other valuable markers. PMID- 15891903 TI - Meningioma-like tumor of the thyroid: a previously undescribed variant of follicular adenoma. PMID- 15891904 TI - Epithelioid solitary fibrous tumor in the ischioanal fossa. PMID- 15891905 TI - Expression of cyclins, p53, and Ki-67 in cervical squamous cell carcinomas: overexpression of cyclin A is a poor prognostic factor in stage Ib and II disease. AB - We previously reported the overexpression of cyclins in uterine cervical carcinoma; however, their clinicopathological significance remained undetermined. In the present study, we examined the immunohistochemical expression of cyclins (D1, E, A, B1), p53 and Ki-67 in squamous cell carcinoma (stage Ib+II; 80 cases, stage III+IV; 23 cases). Correlations between the expression of cyclins and clinicopathological parameters and patient survival were statistically evaluated. The results indicated that in the normal squamous epithelium, the expression of cyclins and Ki-67 was sporadically observed in the parabasal layer. Of the 103 cervical carcinomas, overexpression of cyclins D1, E, A, B1 and p53 was observed in 13 (13%), 23 (22%), 25 (24%), 18 (18%) and 23 (22%) cases, respectively, with a slight predominance in advanced stage tumors. The expression of cyclin D1, E, A and p53 significantly correlated with that of Ki-67 (Spearman's rank correlation). Univariate and multivariate analyses revealed that lymph node metastasis and cyclin A overexpression were independent prognostic factors for unfavorable outcomes in stage Ib+II patients. These findings suggest that the overexpression of various cyclins is involved in the acquisition of the vigorous growth potential of cervical carcinoma cells, and that cyclin A is an independent prognosticator of cervical carcinoma in early stages. PMID- 15891906 TI - Localization of tissue transglutaminase and N (epsilon)-(gamma) -glutamyl lysine in duodenal cucosa during the development of mucosal atrophy in coeliac disease. AB - Expression and transamidation activity of tissue transglutaminase (tTG) may be involved in the morphological modifications leading to the mucosal atrophy observed in coeliac disease (CD). We aimed to investigate the localization of tTG within the duodenal mucosa during the development of villous atrophy. The localization and level of expression of N epsilon-(gamma-glutamyl) lysine isopeptides which could reflect the transamidation activity of tTG were also analyzed. tTG and N epsilon-(gamma-glutamyl) lysine were localized using an immunohistochemical technique on duodenal biopsies obtained from 75 patients with CD and 51 subjects with normal mucosa (control group). The number of cases displaying tTG-expressing cells in the basement membrane and lamina propria was significantly higher in CD patients than in the control group. Moreover, the intensity of tTG staining in these areas was higher in CD. In contrast, the number of biopsies with tTG-expressing enterocytes was significantly lower in CD than in the control group. There was no difference in N epsilon-(gamma-glutamyl) lysine between the two populations. Tissue transglutaminase was differently expressed in the various areas of the mucosa according to the stage of atrophy, whereas the localization and the intensity of the labelling of N epsilon-(gamma glutamyl) lysine isopeptides did not show any modification. The preferential localization in the basement membrane and lamina propria may reflect the involvement of tTG in the development of mucosal atrophy in CD. PMID- 15891907 TI - Adhesion activity of fetal gonadal cells to EGF and discoidin domains of milk fat globule-EGF factor 8 (MFG-E8), a secreted integrin-binding protein which is transiently expressed in mouse early gonadogenesis. AB - MFG-E8, a secreted integrin-binding protein, consists of two EGF domains containing a RGD motif and two discoidin domains. In mouse embryogenesis, MFG-E8 is highly expressed in gonadal stromal cells near mesonephros at 11.5-12.5 dpc, but its function in gonadogenesis has not been characterized. To clarify a possible role of MFG-E8 in developing gonads, we analyzed the adhesion activity of 10.5-15.5 dpc gonadal cells to recombinant proteins of EGF or discoidin domains of MFG-E8. In EGF-coated wells, the gonadal cells at 11.5-12.5 dpc revealed a significantly higher adhesion activity as compared to those at 10.5 and 15.5 dpc, while discoidin domains showed a constant number of the adhered cells throughout these stages. To identify the adhesive cells of 11.5-dpc gonads, immunohistochemistry with anti-SF1/Ad4Bp antibody (a specific marker for supporting, steroidogenic, and coelomic epithelial cells) and staining for alkaline phosphatase (a germ cell marker) were carried out. As a result, EGF domains, as well as discoidin domains, were capable of binding to all three groups of SF1/Ad4Bp-positive and negative somatic cells, and germ cells of 11.5 dpc gonads. These findings therefore suggest that MFG-E8 mediates the cell-to cell interaction among several somatic cell types and germ cells in mouse early gonadogenesis. PMID- 15891908 TI - Asymmetric cell divisions are concentrated in the dermomyotome dorsomedial lip during epaxial primary myotome morphogenesis. AB - To determine if somitic stem cell pools could be identified by an intrinsic difference in mitotic behaviour, the orientation of mitoses in the dermomyotome epithelium was analysed. We describe a concentration of apico-basal mitoses within the dermomyotome dorsomedial lip (DML). The occurrence of apico-basal divisions is closely associated with asymmetric localisation of the notch pathway factor numb, allowing description of such divisions as asymmetric. In contrast, planar divisions, occurring in the plane of the epithelium, are symmetric. Further, we show that the DML environmental niche is sufficient to promote numb expression in epaxial dermomyotome tissue that does not normally express this factor. These data provide, for the first time, a non-retrospective tracing analysis of the mechanism by which the DML fulfils the stem-cell pool role it plays during epaxial primary myotome morphogenesis. PMID- 15891909 TI - Wnts and the neural crest. AB - The neural crest is a multipotent tissue that originates between the neural epithelium and non-neural ectoderm, which can develop into numerous cell types, including neurons, glia, pigment cells, smooth muscle, cartilage and bone. Work in a variety of animal models has shown that a number of signalling factors are necessary for the induction, delamination and differentiation of neural crest cells. However one family of proteins, the Wnts, shows an overriding influence on this tissue. Here we review recent studies that pinpoint specific roles that Wnts play in the development of the neural crest. PMID- 15891910 TI - Retrotransposon populations of Vicia species with varying genome size. AB - The (non-LTR) LINE and Ty3-gypsy-type LTR retrotransposon populations of three Vicia species that differ in genome size (Vicia faba, Vicia melanops and Vicia sativa) have been characterised. In each species the LINE retrotransposons comprise a complex, very heterogeneous set of sequences, while the Ty3-gypsy elements are much more homogeneous. Copy numbers of all three retrotransposon groups (Ty1-copia, Ty3-gypsy and LINE) in these species have been estimated by random genomic sequencing and Southern hybridisation analysis. The Ty3-gypsy elements are extremely numerous in all species, accounting for 18-35% of their genomes. The Ty1-copia group elements are somewhat less abundant and LINE elements are present in still lower amounts. Collectively, 20-45% of the genomes of these three Vicia species are comprised of retrotransposons. These data show that the three retrotransposon groups have proliferated to different extents in members of the Vicia genus and high proliferation has been associated with homogenisation of the retrotransposon population. PMID- 15891911 TI - Identification and distribution of sequences having similarity to mitochondrial plasmids in mitochondrial genomes of filamentous fungi. AB - Mitochondrial plasmids are autonomously replicating genetic elements commonly associated with fungal and plant species. Analysis of several plant and fungal mitochondrial genomes has revealed regions that show significant homology to mitochondrial plasmids, suggesting that plasmids have had a long-term association with their mitochondrial hosts. To assess the degree to which plasmids have invaded fungal mitochondrial genomes, BLAST search parameters were modified to identify plasmid sequences within highly AT-rich mtDNAs, and output data were parsed by E value, score, and sequence complexity. High scoring hits were evaluated for the presence of shared repetitive elements and location within plasmids and mtDNAs. Our searches revealed multiple sites of sequence similarity to four distinct plasmids in the wild-type mtDNA of Neurospora crassa, which collectively comprise more than 2% of the mitochondrial genome. Regions of plasmid similarity were not restricted to plasmids known to be associated with senescence, indicating that all mt plasmids can potentially integrate into mitochondrial DNA. Unexpectedly, plasmid-related sequences were found to be clustered in regions that have disproportionately low numbers of PstI palindromic sequences, suggesting that these repetitive elements may play a role in eliminating foreign DNA. A separate class of GC-rich palindromes was identified that appear to be mobile, as indicated by their occurrence within regions of plasmid homology. Sites of sequence similarity to mitochondrial plasmids were also detected in other filamentous fungi, but to a lesser degree. The tools developed here will be useful in assessing the contribution plasmids have made to mitochondrial function and in understanding the co-evolution of mitochondrial plasmids and their hosts. PMID- 15891912 TI - Identification by suppression subtractive hybridization of genes that are differentially expressed between near-isogenic maize lines in association with sugarcane mosaic virus resistance. AB - The molecular mechanisms underlying the development and progression of sugarcane mosaic virus (SCMV) infection in maize are poorly understood. A study of differential expression was conducted to identify genes involved in resistance to SCMV. In this study, we combined suppression subtractive hybridization and macroarray hybridization to identify genes that are differently expressed in the near isogenic lines F7+ (SCMV resistant) and F7 (susceptible). Altogether, 302 differentially expressed genes were identified in four comparisons based on constitutively expressed and inducible genes, and on compatible and incompatible plant-virus interactions. Apart from genes related to metabolism, most of the functionally classified genes identified belonged to three pathogenesis-related categories: cell rescue, defense, cell death and ageing, signal transduction, and transcription. The latter three groups accounted for 56-66% of the genes classified. Some 19% (60 of 302) of the identified genes had previously been assigned to 29 bins distributed over all ten maize chromosomes. Among the mapped genes, 31% (18 of 58) were located within the Scmv2 and Scmv1 regions on chromosomes 3 and 6, respectively, which have been associated with resistance to SCMV. Promising candidate genes for Scmv1 have been identified, such as AA661457 (receptor-like kinase Xa21-binding protein 3). The implications of the genomic distribution of differentially expressed genes identified by this isogenic comparison are discussed in the context of breeding for resistance. PMID- 15891913 TI - Subcellular localization of spastin: implications for the pathogenesis of hereditary spastic paraplegia. AB - Hereditary spastic paraplegia (HSP) is a group of clinically and genetically heterogeneous diseases characterized by neuronal degeneration that is maximal at the distal ends of the longest axons of the central nervous system. The most common cause of autosomal dominant HSP is mutation of a novel gene encoding spastin, a protein whose function is still being elucidated. One clue concerning spastin function is its intracellular localization. Here, we describe a novel anti-spastin antiserum designed to a unique epitope contained within all splicing isoforms. The antiserum exhibits specific immunostaining of recombinant spastin in intact, fixed cells. Using this reagent, we find that endogenous spastin is located at the centrosome in a variety of cell types at all points in the cell cycle. This localization is resistant to microtubule disruption, suggesting that spastin may be an integral centrosomal protein. In addition to the centrosome, spastin also localizes at discrete focal regions along the axons of primary cultured neurons. These data lend additional support to the emerging hypothesis that spastin plays a role in microtubule dynamics, with a crucial role in microtubule organization. PMID- 15891915 TI - Involvement of adenovirus in clinical mononucleosis-like syndromes in young children. AB - Although Epstein-Barr virus (EBV) commonly causes infectious mononucleosis (IM) or IM-like syndromes, other agents can be implicated. In this study, viral and parasitic screening was performed to determine the etiological agent of pediatric IM-like syndromes in 38 children. Adenovirus was the agent most frequently detected (47.3%), followed by EBV (31.5%) and cytomegalovirus (2.6%). Although the statistically significant difference between viral detection rates observed in patients who fulfilled clinical and hematological criteria and detection rates in those who presented clinical symptoms only (91.6% vs. 64.3%) indicates that hematological abnormalities are common in viral IM-like syndromes, the existence of syndromes of viral etiology without hematological criteria cannot be discarded. A further analysis showed an absence of lymphocytosis in adenovirus infections as well as a low number (14.3%) of EBV infections associated with increased neutrophils. These data suggest the usefulness of appropriate virological techniques for the detection of adenovirus in pediatric IM-like syndromes. PMID- 15891914 TI - Clostridium clostridioforme: a mixture of three clinically important species. AB - Clostridium clostridioforme shows much variability in phenotypic and antimicrobial susceptibility tests, suggesting it may be more than a single species even though all strains share unique morphology. This study was designed to determine if there are multiple species and, if so, to demonstrate the differences that exist between them. A total of 107 strains of C. clostridioforme were investigated by sequencing of the 16S rRNA gene, phenotypic studies, and antimicrobial susceptibility testing. In addition, clinical data from patients whose infections yielded an organism identified as C. clostridioforme was reviewed. Data from the above studies revealed three principal species in what has been called C. clostridioforme: Clostridium bolteae, C. clostridioforme, and Clostridium hathewayi. Each species may be distinguished by certain phenotypic tests. All three species were involved in infections, including bacteremia. C. clostridioforme appears to be associated with more serious or invasive human infections than the other two species in the group. Resistance to penicillin G is common and is due to beta-lactamase production. Resistance to clindamycin and moxifloxacin is also seen. The three species differ in terms of virulence and antimicrobial resistance. "C. clostridioforme" actually represents three distinct species that are different in terms of 16S rRNA sequences, phenotypic characteristics, and antimicrobial susceptibility. It is important for microbiology laboratories to distinguish between these species and for clinicians to be aware of the differences between them. PMID- 15891916 TI - Prospective evaluation of a real-time PCR assay for detection of group B streptococci in vaginal swabs from pregnant women. PMID- 15891917 TI - Comparison of genotypes of methicillin-resistant and methicillin-sensitive Staphylococcus aureus in Finland. AB - The frequency of horizontal transfer of the staphylococcal cassette chromosome mec to methicillin-susceptible Staphylococcus aureus is unknown. In order to gain more information regarding this frequency in Finland, the genotypes of 299 clinical methicillin-sensitive Staphylococcus aureus isolates were compared to representatives of 24 epidemic methicillin-resistant Staphylococcus aureus genotypes. Sixty-eight percent of the methicillin-sensitive isolates had a genotype similar to eight of the epidemic methicillin-resistant strains. The remaining isolates (32%) showed 22 different genotypes. The results indicate that, in Finland, several methicillin-sensitive Staphylococcus aureus genotypes may have acquired the staphylococcal cassette chromosome mec. PMID- 15891918 TI - Value of PET-CT fusion imaging in avoiding potential pitfalls in the interpretation of 18F-FDG accumulation in the distal oesophagus. PMID- 15891919 TI - Haemodynamic effects of dual-chamber pacing versus ventricular pacing during a walk test in patients with depressed or normal left ventricular function. AB - PURPOSE: Dual-chamber rate-modulated pacing provides haemodynamic benefits compared with ventricular pacing at rest, but it is unclear whether this also holds true during physical exercise in patients with heart failure. This study assessed the haemodynamic response to a walk test during dual-chamber pacing and ventricular pacing in patients with depressed or normal left ventricular (LV) function. METHODS: Twelve patients with an LV ejection fraction <50% and 11 patients with an LV ejection fraction >or=50% underwent two randomised 6-min walk tests under dual-chamber rate-modulated pacing and ventricular pacing at a fixed rate of 70 beats/min. All patients had a dual-chamber pacemaker implanted for complete heart block. LV function was monitored by a radionuclide ambulatory system. RESULTS: In patients with depressed LV function, the change from dual chamber pacing to ventricular pacing induced a decrease in end-systolic volume at the peak of the walk test (P<0.05), with no difference in end-diastolic volume. As a consequence, higher increases in LV ejection fraction (P<0.0001) and stroke volume (P<0.01) were observed during ventricular pacing. No difference in cardiac output was found between the two pacing modes. In patients with normal LV function, the change from dual-chamber pacing to ventricular pacing induced a significant decrease in cardiac output (P<0.005 at rest and P<0.05 at the peak of the walk test). CONCLUSION: Compared with dual-chamber rate-modulated pacing, ventricular pacing improves cardiac function and does not affect cardiac output during physical activity in patients with depressed LV function, whereas it impairs cardiac output in those with normal function. PMID- 15891920 TI - Fatty marrow conversion of the proximal femoral metaphysis in transient bone marrow edema syndrome. AB - INTRODUCTION: In the proximal femoral metaphysis, hematopoietic marrow is predominant during the adult stage of life. The conversion of hematopoietic marrow to fatty marrow in the proximal femoral metaphysis has been suggested as an etiologic factor of ischemia in the pathogenesis of femoral head osteonecrosis. To determine whether the chronology of fatty marrow conversion of the proximal femoral metaphysis is related to transient bone marrow edema syndrome of the hip, a case control study was conducted on 10 patients with the disease. MATERIALS AND METHODS: There were 8 men and 2 women with a mean age of 33 years (range 19-45 years). The 10 patients were matched with 20 controls for gender and age (5-year range). T1-weighted MRI scans of their hips were reviewed. Marrow of the greater trochanter becomes fatty before puberty, and thus, the greater trochanter can be used as a built-in control. The signal intensity of the proximal femoral metaphysis was compared to that of the greater trochanter. RESULTS: In all patients, the signal intensity of the proximal femoral metaphysis was isointense (fatty marrow) relative to that of the greater trochanter. In control subjects, the signal intensity was isointense in 8 (40%) and hypointense (hematopoietic marrow) in 12 (60%) (p < 0.05). CONCLUSION: The current study shows that the proximal femoral metaphysis is predominantly fatty in transient bone marrow edema syndrome. The conversion of hematopoietic to fatty marrow is known to correlate with decreases in intramedullary blood flow. Thus, the current study suggests that an ischemia of the proximal femur secondary to fatty marrow conversion of the proximal femoral metaphysis might be a cause of transient bone marrow edema syndrome of the hip. PMID- 15891921 TI - Palmar locking plate for treatment of unstable dorsal dislocated distal radius fractures. AB - INTRODUCTION: This study investigates the clinical and radiological results of open reduction and internal fixation of unstable distal radial fractures using a palmar placed interlocking plate system with no substitutes for treatment of the dorsal metaphyseal fracture void. MATERIALS AND METHODS: Nineteen patients with a mean age of 61 years (range 49-78 years) and a dorsally displaced distal radial fracture were treated using a palmar placed 2.4 mm interlocking plate system. All 17 women and 2 men were followed up according to the study protocol. Standardised radiographs in anteroposterior and lateral projection were performed before reduction and at 2, 6, 12 and 24 weeks after surgery. Radiological measurements included palmar tilt, radial inclination and radial shortening as defined by ulnar variance and intra-articular steps. Clinical assessment included active range of motion (ARM) of the wrist, pain according to a visual analogue scale (VAS), grip power, working ability, Disability of Arm, Shoulder and Hand-Score (DASH Score) and Mayo Wrist Score. RESULTS: At final follow-up after a minimum of 11 months, the palmar tilt was +1 degrees , radial inclination 19.8 degrees and ulnar variance 1.0 mm. Mean loss of palmar tilt measured 1.9 degrees , radial inclination 1.3 degrees and ulnar variance 0.4 mm. Mean wrist extension measured 58 degrees , wrist flexion 41 degrees , pronation 83 degrees and supination 84 degrees . The Mayo Wrist Score showed 21% excellent, 58% good, 16% fair and 5% poor results. CONCLUSION: The treatment of unstable, dorsally dislocated, distal radius fractures AO types A3, C1 and C2, with a palmar placed 2.4 mm interlocking plate system and without additional treatment of the metaphyseal defects showed good radiological and functional results with minimal loss of reduction. PMID- 15891922 TI - Distribution of substance-P nerves inside the infrapatellar fat pad and the adjacent synovial tissue: a neurohistological approach to anterior knee pain syndrome. AB - INTRODUCTION: The purpose of the study was to determine the distribution and number of nerves inside the infrapatellar fat pad and the adjacent synovium, in particular with regards to nociceptive substance-P nerves. MATERIALS AND METHODS: The infrapatellar fat pad of the knee was resected from 21 patients (4 male, 17 female, mean age 69 years) during the course of standard total knee arthroplasty operations performed in our clinic. The fat pad was dissected into five standardized segments, fixed in formalin and embedded in paraffin. Immunohistochemical techniques using antibodies against S-100 protein and substance-P (SP) were employed to determine and specify the nerves. RESULTS: Studying all the detectable nerves present in 50 observation fields (200-fold magnification), we found an average of 106 S-100 versus 25 SP nerves (24%) in the synovium and 27 S-100- versus 7 SP nerves (26%) in the interior of the fat pad. The total nerve count was significantly (P < 0.001) higher in the synovium than in the fat pad for both marker types. The number of S-100 nerves was significantly (P < 0.05) higher in the central and lateral segments of the fat pad, while SP nerves were equally distributed throughout all segments of the fat pad. SP nerves were significantly more frequently associated with blood vessels inside the fat pad (43%, P < 0.05) than in the synovial tissue (28%). CONCLUSION: The occurrence and distribution of SP nerves inside the infrapatellar fat pad suggest a nociceptive function and a neurohistological role in anterior knee pain syndrome. The data support the hypothesis that a neurogenous infection of the infrapatellar fat pad could contribute to anterior knee pain syndrome. PMID- 15891923 TI - Mycophenolate mofetil in steroid/cyclosporine-dependent/resistant nephrotic syndrome. AB - Attempts to minimize the effects of prolonged steroid use in steroid-dependent nephrotic syndrome (SDNS) and the need to overcome steroid resistance (SRNS) justifies immunosuppressant therapy. We report our experience in a cohort of patients with SD/SRNS during the administration of mycophenolate mofetil (MMF) in a prospective protocol initiated in January 2001. Twenty-six children with idiopathic nephrotic syndrome were included (21 steroid dependent and 5 steroid resistant), whose response did not change after sequential treatment with cyclophosphamide (CPM) and cyclosporine (CsA). Histopathologic patterns were: 11 minimal change disease (MCD), 1 diffuse mesangial proliferation (DMP), 13 focal segmental glomerulosclerosis (FSGS) and membranous 1 glomerulonephritis (MGN). The median age of onset of NS was 2.8 years (range 1.2-12.5), and treatment with MMF was performed at a median age of 11.4 years (range 5-17) with an initial dose of 600 mg/m(2)/12 h, adjusted to maintain levels of mycophenolic acid (MPA) at 2.5-5 mcg/ml. The planned duration of study to assess treatment efficacy was 6 months. The mean MMF dose required was 624 (SD=136) mg/m(2)/12 h (range 415-970), which maintained mean C(0)-MPA levels of 2.9 (SD=1.17) mcg/ml (range 1.2-5.9 mcg/ml). In the five patients with SRNS, only one achieved complete remission. In the patients with SDNS, steroid sparing was achieved in 15 and 9 remained in remission on MMF monotherapy. Withdrawal of MMF resulted in immediate relapse in 47%. In our study, MMF was a useful immunosuppressant due to its fewer undesirable effects and similar efficacy to other drugs used. It appears effective for the maintenance of remission in SDNS patients, with a response similar to that of CsA. PMID- 15891924 TI - Premedication during micturating cystourethrogram to achieve sedation and anxiolysis. AB - Micturating cystourethrogram (MCUG) is an imaging technique indicated in the diagnosis and follow-up of many diseases. We investigated the reliability and the efficacy of midazolam and chloral hydrate in sedation and anxiolysis during micturating cystourethrogram. Fifty-three children of similar ages (39 girls, 14 boys, mean age of 5.8+/-3.5 years) were randomized to midazolam (n=17), chloral hydrate (n=18) and control groups (n=18). Oral midazolam 0.6 mg/kg or chloral hydrate 25 mg/kg or saline were administered to the study groups 15-30 min prior to the urinary catheterization. Brietkopf and Buttner, Frankl and Houpt scales and Spielberger's State Anxiety Inventory and parent's impressions were used to assess the level of sedation and anxiety. The Brietkopf and Buttner classification of emotional status and Houpt behavior rating scale demonstrated a significantly better emotional status and sedation in the midazolam group when compared to controls (P=0.01 and P=0.018, respectively). The catheterization was described as a more unpleasant and distressing event by the parents of the control and the chloral hydrate groups when compared to the parents of the midazolam group (P<0.05). Bladder capacity and frequency of detection of residual urine were not statistically different between the three study groups (P>0.05). Vital signs did not change significantly in any child. Sedation with midazolam does not have adverse effects on the results of micturating cystourethrogram, while it reduces the discomfort in children undergoing this radiological technique. PMID- 15891925 TI - Metachronous second primary tumours in the aerodigestive tract in patients with early stage head and neck squamous cell carcinomas. AB - Early-stage head and neck carcinomas can usually be controlled with the appropriate treatment. In these patients, the long-term prognosis mainly depends on second metachronous malignancies, frequently in the aerodigestive tract. Our study aims to identify risk factors for the appearance of second tumours in this group of patients with early head and neck cancer. Of 949 patients included in the study, 189 (20%) developed a metachronous second primary malignancy, most frequently in the aerodigestive tract. Independent risk factors associated with second tumours were heavy alcohol use and the location of the index tumour in the oropharynx. Compared to non-drinkers, heavy drinkers (>80 g/day) presented a 1.8 times higher risk of a second tumour (CI 95%: 1.01-3.50). Patients with oropharyngeal tumours had a 2.15-higher risk than patients with oral cavity tumours (CI 95%: 1.03-4.47). Recursive partitioning analysis was used to characterise two risk groups for second tumours. The low-risk group included patients over 75 years and patients with low levels of carcinogen use. It comprised 171 patients (18%) with a 5.3% frequency of second tumours. The high risk group accounted for 80% of the patients (n = 778), and the rate of second neoplasms was 16.3%. Classification of the patients according to the mentioned variables allows us to focus follow-up and prevention efforts on high-risk patients. PMID- 15891926 TI - Middle cranial fossa arachnoid cysts causing sensorineural hearing loss. AB - Arachnoid cyst of the middle cranial fossa has never been reported to present with sensorineural hearing impairment. We report the case of a 15-year-old girl who complained of progressive hearing loss on the right side for 4 years. The pure tone audiometry revealed sensorineural hearing loss with an average of 63 dB, and the auditory brain stem response demonstrated absence of all waves on the right side. Magnetic resonance imaging showed giant arachnoid cysts occupying the bilateral middle cranial fossa and extending to the parietal and prepontine areas with the atrophic change of the right facial-acoustic nerve cord and a decrease in signal intensity of the right cochlea and vestibule. The cystoperitoneal shunting procedure was suggested, but the parents hesitated about the surgical treatment. The patient has not received any surgical intervention, and her hearing has been stable for 1 year during the follow-up period. This case represents the first report that sensorineural hearing loss is a possible presenting symptom when arachnoid cysts of the middle cranial fossa are massive. PMID- 15891927 TI - Rhinocerebral mucormycosis: pathways of spread. AB - Rhinocerebral mucormycosis is an invasive, opportunistic fungal infection usually seen in immunocompromised patients, and particularly in the setting of diabetes or immune deficiency. It is assumed that the port of entry is colonization of the nasal mucosa, allowing the fungus to spread via the paranasal sinuses into the orbit. Involvement of the brain and cavernous sinus occurs by way of the orbital apex; therefore, spheno-ethmoidectomy with or without maxillectomy seems to be the definitive method to eradicate this infection. We conducted a prospective study of ten patients with rhinocerebral mucormycosis from February 2000 to April 2004. Rhinocerebral mucormycosis was clinically diagnosed in 11 patients, 10 of whom were included in our study upon histopathological confirmation. Diabetes was the most common underlying disorder seen in nine out of ten patients. In this study, the patients were assessed for predisposing factors, presenting signs and symptoms, sites of extension, the number and sites of surgical debridement, as well as the outcome. Ocular, sinonasal and facial soft tissue involvement was common. Involvement of the pterygopalatine fossa at the time of debridement was evident in all patients. No invasion through the lamina papiracea or the walls of the maxillary sinus was identified. At the time of this communication, six out of ten patients were alive. For the four who died, the causes were hypokalemia, cardiac arrythmia and refractory pneumonia. Pterygopalatine fossa is considered to be the main reservoir for rhinocerebral mucormycosis, and extension into the orbit and facial soft tissues usually follows this route. After proliferation in the nasal cavity, the mucor reaches the pterygo-palatine fossa, inferior orbital fissure and finally the retroglobal space of the orbit, resulting in ocular signs. The facial soft tissues, palate and infratemporal fossa can be infected through connecting pathways from the pterygo-palatine fossa; therefore, debridement of the pterygopalatine fossa seems to be the definitive method of managing this infection. PMID- 15891928 TI - A simple model for shear stress mediated lumen reduction in blood vessels. AB - The goal of this paper is to qualitatively describe the dominant flow characteristics associated with shear stress mediated lumen reduction due to atherosclerotic plaque growth. The approach is to develop rate equations for the reduction of the lumen as a function of the shear stress near the intima wall. Elementary, qualitative, models for fully developed laminar and turbulent flows are employed, leading to nonlinear ordinary differential equations. The model provides a qualitative description of one aspect of the long-term reduction, perhaps taking years, of the lumen due to wall growth. This is useful because of the extreme complexity of long-term experiments. PMID- 15891929 TI - Common mutations of beta-catenin in adamantinomatous craniopharyngiomas but not in other tumours originating from the sellar region. AB - Dysregulation of the Wnt signalling pathway contributes to developmental abnormalities and carcinogenesis of solid tumours. Here, we examined beta-catenin and adenomatous polyposis coli (APC) by mutational analysis in pituitary adenomas (n=60) and a large series of craniopharyngiomas (n=41). Furthermore, the expression pattern of beta-catenin was immunohistochemically analysed in a cohort of tumours and cysts of the sellar region including pituitary adenomas (n=58), craniopharyngiomas (n=57), arachnoidal cysts (n=8), Rathke's cleft cysts (n=10) and xanthogranulomas (n=6). Whereas APC mutations were not detectable in any tumour entity, beta-catenin mutations were present in 77% of craniopharyngiomas, exclusively of the adamantinomatous subtype. All mutations affected exon 3, which encodes the degradation targeting box of beta-catenin compatible with an accumulation of nuclear beta-catenin protein. In addition, a novel 81-bp deletion of this exonic region was detected in one case. Immunohistochemical analysis confirmed a shift from membrane-bound to nuclear accumulation of beta-catenin in 94% of the adamantinomatous tumours. Aberrant distribution patterns of beta catenin were never observed in the other tumour entities under study. We conclude that beta-catenin mutations and/or nuclear accumulation serve as diagnostic hallmarks of the adamantinomatous variant, setting it apart from the papillary variant of craniopharyngioma. PMID- 15891930 TI - Adductor insertion avulsion syndrome, "thigh splints": relevance of radiological follow-up. AB - We present a case of chronic osteomyelitis in a 13-year-old girl which was originally diagnosed as adductor insertion avulsion syndrome ("thigh splints") on the basis of the clinical presentation, patient history, initial radiographs and MRI examination. However, at follow-up with persistent pain and altered radiographic and MRI appearances, surgical biopsy was indicated. Histopathological findings confirmed a bone abscess. This case underlines the necessity of clinical follow-up and imaging in certain patients with apparent thigh splints. PMID- 15891931 TI - Spondylocarpotarsal synostosis syndrome (with a posterior midline unsegmented bar). AB - Spondylocarpotarsal synostosis syndrome (SSS) is characterised by malsegmentation of the thoracic spine and carpal/tarsal fusions. A unilateral or bilateral unsegmented bar may be present in the thoracic spine. Presenting clinical signs are congenital scoliosis early in life, and shortening of the trunk with scoliosis and/or lordosis in older children. We report a 13-year-old girl with SSS and a midline unsegmented bar running along the spinal processes of T3 to L2 and extending into the posterior vertebral elements. PMID- 15891932 TI - [Problem-based learning for surgery. Increased motivation with less teaching personnel?]. AB - BACKGROUND: Traditional teacher-centred education strategies often do not meet the needs and student abilities of adult learning. The introduction of small learning groups requires considerable increases in staff. Problem-based learning could increase the motivation to acquire knowledge but without being as staff intensive. METHODS: Medical students (n=98) in their fourth clinical semester were randomly assigned to either a structured course (SC) or problem-based learning (PBL) for surgery. Their motivation and acceptance of the courses were recorded at the end of term in anonymous questionnaires using Likert scales, with scores ranging from 1 (very good) to 6 (unsatisfactory). RESULTS: Both course structure and the teachers received much better ratings from the PBL students (P<0.01 each). The motivation of students to deal with surgical problems beyond the course material was significantly higher after PBL, with 1.8 (0.7), than after the structured course with 3.1 (1.2) (P<0.01). The overall rating was substantially worse for the structured course, with 3.1 (1.2) than for PBL at 1.4 (0.6) (P<0.01). CONCLUSIONS: Problem-based learning in the surgical curriculum increases student acceptance and motivation with little demand on staff. It should be increasingly implemented. PMID- 15891933 TI - The ability of acidic pH, growth inhibitors, and glucose to increase the proton motive force and energy spilling of amino acid-fermenting Clostridium sporogenes MD1 cultures. AB - Clostridium sporogenes MD1 grew rapidly with peptides and amino acids as an energy source at pH 6.7. However, the proton motive force (Deltap) was only -25 mV, and protonophores did not inhibit growth. When extracellular pH was decreased with HCl, the chemical gradient of protons (ZDeltapH) and the electrical membrane potential (DeltaPsi) increased. The Deltap was -125 mV at pH 4.7, even though growth was not observed. At pH 6.7, glucose addition did not cause an increase in growth rate, but DeltaPsi increased to -70 mV. Protein synthesis inhibitors also significantly increased DeltaPsi. Non-growing, arginine-energized cells had a DeltaPsi of -80 mV at pH 6.7 or pH 4.7, but DeltaPsi was not detected if the F1F0 ATPase was inhibited. Arginine-energized cells initiated growth if other amino acids were added at pH 6.7, and DeltaPsi and ATP declined. At pH 4.7, ATP production remained high. However, growth could not be initiated, and neither DeltaPsi nor the intracellular ATP concentration declined. Based on these results, it appears that C. sporogenes MD1 does not need a large Deltap to grow, and Deltap appears to serve as a mechanism of ATP dissipation or energy spilling. PMID- 15891934 TI - Production of red pigments by the insect pathogenic fungus Cordyceps unilateralis BCC 1869. AB - Production of red pigments (naphthoquinones) by the insect pathogenic fungus Cordyceps unilateralis BCC 1869 was investigated in this study. Cultivation conditions, including temperature, intitial pH of medium, and aeration, were optimised to improve the yield of total naphthoquinones in shake-flask culture of C. unilateralis. The highest yield of total naphthoquinones (3 g L-1) was obtained from a 28-day culture grown in potato dextrose broth with an initial pH of 7.0, at 28 degrees C with shaking-induced aeration at 200 rpm. An extraction process for isolation of the targeted naphthoquinone, 3,5,8-trihydroxy-6-methoxy 2-(5-oxohexa-1,3-dienyl)-1,4-naphthoquinone (3,5,8-TMON), from a culture of C. unilateralis, was also developed. The yield of 3,5,8-TMON obtained was about 1.2 g L-1 or 40% of total naphthoquinones. The stability of 3,5,8-TMON was very high, even upon exposure to strong sunlight (70,000 lx), high temperature up to 200 degrees C, and acid and alkali solutions at concentrations of 0.1 M. PMID- 15891937 TI - Prognosis of hepatocellular carcinoma after hepatectomy in patients with renal dysfunction. AB - We evaluated the effect of renal dysfunction on the prognosis of hepatocellular carcinoma in 224 patients who underwent hepatic resection. Survival was compared between patients with (57 patients) and without renal dysfunction (116 patients), using a creatinine clearance 75% ventricular thickness comprised of noncompacted area with recess adjacent to tricuspid valve) was seen in 6 of 14 hearts. One patient had a sibling with pulmonary stenosis, but there was no other known familial cardiomyopathy. Endocardial fibroelastosis was a characteristic histological feature, as well as anastomosing or polypoid endocardial trabeculations, which resulted in staghorn shaped, endocardial-lined recesses. There was a high rate of other cardiac anomalies, which often coexisted and were not clearly related to the LVNC, present in 8 cases (nonisolated LVNC): ventricular septal defect (4/14), anomalous venous pulmonary veins (1/14), coronary ostial stenosis (1/14), histiocytoid cardiomyopathy (1/14), polyvalvar dysplasia (2/14), and pulmonary stenosis (2/14). In the 6 isolated LVNC, there were 2 malformed atrioventricular valves (1 mitral and 1 tricuspid), which appeared part of the ventricular maldevelopment. There were no differences in histological or gross patterns of the noncompacted regions between the isolated and nonisolated LVNC. LVNC is frequently associated with other cardiac defects, especially when causing sudden death in infants and children. A clear-cut morphological distinction between "isolated" and "secondary" LVNC was not apparent. The pathologist should be aware of the entity because the diagnosis is often first established at autopsy. PMID- 15892003 TI - Subepidermal cleft formation as a diagnostic marker for cutaneous malignant melanoma. AB - Cleft formation has been postulated as a clue to the histopathological diagnosis of malignant melanoma (MM). The frequency and reliability of clefts as a diagnostic criterion remain to be determined. We reviewed 503 cases of histologically proven MM searching for clefting between the epidermal layer and underlying MM. Cleft was defined as a separation of at least 0.3 mm in length. Conspicuous cleft formation was present in 120 (24%) of 503 MMs. The presence of clefts was not associated with age or sex of the patients, but showed a slight predilection for the back, a slightly higher prevalence in superficial spreading type of MM and for tumors with a Breslow thickness between 1 and 2 mm. Morphologically, clefts could be separated in 3 different types: linear (37.5%), single-nest (10.9%), and multi-nest (51.6%). In comparison, among 939 benign melanocytic lesions including 100 Spitz or Reed nevi, only 9 exhibited clefts longer than 0.3 mm (< 1%). One was an atypical compound nevus; all others were Spitz nevi, with the majority exhibiting an arched morphology above 1 or 2 large round nests. The relative frequency of cleft formation allowed a highly significant differentiation between MM and benign melanocytic lesions. Clefts are a reliable diagnostic criterion in favor of MM. PMID- 15892004 TI - Aberrant CpG island hypermethylation profile is associated with atypical and anaplastic meningiomas. AB - Hypermethylation of promoter CpG islands is a common epigenetic event in a variety of human cancers. The aim of this study was to investigate whether promoter hypermethylation of cancer-related genes is involved in the development and progression of meningiomas. The methylation status at the promoter region of 10 cancer-related genes was examined by methylation-specific polymerase chain reaction in a cohort of 48 meningiomas including 16 benign, 19 atypical, and 13 anaplastic variants. The relationship of promoter hypermethylation and transcriptional silencing was determined by treatment of cells with demethylating agent 5-aza-2'-deoxycytidine followed by reverse transcription-polymerase chain reaction. Our results showed that 50% (24/48) of meningiomas exhibited promoter hypermethylation in at least one of the genes but not in normal leptomeninges, indicating that aberrant hypermethylation is tumor-specific. Promoter hypermethylation was detected in glutathione S -transferases P1 at 27%, thrombospondin-1 at 15%, retinoblastoma 1 at 10%, cyclin-dependent kinase inhibitor 2A at 10%, O 6 -methylguanine-DNA methyltransferase at 6%, and death associated protein kinase 1, von Hippel-Lindau, p14 ARF , and cyclin-dependent kinase inhibitor 2B, each at 4%. No promoter hypermethylation was detected in the tissue inhibitor of metalloproteinase 3 gene. Treatment of IOMM-Lee meningioma cell line with 5-aza-2'-deoxycytidine restored expression of O 6 -methylguanine DNA methyltransferase and death-associated protein kinase 1, providing evidence that promoter hypermethylation contributes to transcriptional silencing. The frequencies of methylation of any single gene in benign, atypical, and malignant meningiomas were 6% (1/16), 74% (14/19), and 69% (9/13), respectively. Of 48 tumors, 13 (27%) showed that concurrent hypermethylation of two or more genes studied were of atypical or anaplastic type. Statistical analysis revealed that the incidence of promoter hypermethylation of any single gene, of multiple genes, or of glutathione S -transferase P1 was significantly associated with atypical and anaplastic meningiomas ( P < .0001, P = .004, and P = .004, respectively). In conclusion, this study demonstrates that aberrant hypermethylation profile is associated with atypical and anaplastic meningiomas, suggesting that epigenetic change may be involved in malignant progression of meningiomas. PMID- 15892005 TI - Intraabdominal cystic lymphangiomas obscured by marked superimposed reactive changes: clinicopathological analysis of a series. AB - Cystic (or cavernous) lymphangiomas are uncommon tumors that most often occur in the head and neck, axilla, or groin of young children but are detected occasionally in adulthood at various other anatomic sites. When arising in the abdomen, cystic lymphangiomas may present with acute abdominal pain. We have encountered examples of mesenteric and retroperitoneal cystic lymphangiomas associated with such marked superimposed reactive and inflammatory changes that their lymphatic nature is obscured, a situation that is not widely recognized. To further characterize these lesions, 7 abdominal lymphangiomas associated with florid reactive changes were retrieved from the authors' consultation files. There were 5 female patients and 2 male patients (median age, 42 years; range, 1 month to 51 years). Five cases presented in adulthood. Tumor size ranged from 8 to 20 cm (median, 15 cm). Three tumors arose in the mesentery of the small intestine and 4 arose in the retroperitoneum (one of which also involved the posterior mediastinum). Three patients presented with a short history of abdominal pain. Radiological studies revealed large cystic or solid masses; clinical differential diagnoses included sarcoma (2 cases), enteric duplication cyst (2 cases), and cystic tumor not otherwise specified. Grossly, the tumors were generally multiloculated cystic masses associated with areas of fat necrosis and hemorrhage. The cysts often contained thick, gelatinous, or milky fluid. Histologically, all cases showed extensive areas of granulation tissue, most also including a floridly cellular reactive myofibroblastic proliferation, obscuring the lymphatic nature of the lesion. Two cases contained extensive areas of xanthogranulomatous inflammation. In foci where the underlying lesion could be discerned, the tumors were composed of cystically dilated lymphatic spaces, some of which were partially invested by a layer of smooth muscle and were associated with occasional lymphoid aggregates. The lymphatic spaces contained either clear fluid or large numbers of foamy macrophages. The lymphatic endothelial cells lining the cystic spaces were generally attenuated with no cytological atypia. One case showed features of a complex vascular malformation with a predominant component of cavernous lymphangioma. By immunohistochemistry, in all cases, the endothelial cells lining the dilated lymphatic spaces were positive for CD31 and D2-40, 4 of 7 were positive for CD34, and all were negative for keratin. Clinical follow-up information was available for 4 patients (median, 26 months; range, 22 36 months): 3 patients had no evidence of recurrence and 1 patient was asymptomatic with radiographic evidence of minimal persistent disease. In summary, some intraabdominal lymphangiomas have a tendency to induce marked reactive and inflammatory changes in the surrounding tissues, often obscuring their nature and occasionally leading to the clinical impression of a malignant tumor. Awareness of this unusual occurrence will lead to the correct diagnosis. PMID- 15892006 TI - Ovarian serous cystadenoma with mural nodules of genital rhabdomyoma. AB - We present an extremely rare case of ovarian serous cystadenoma with mural nodules of rhabdomyoma. The patient, a 48-year-old woman, was admitted to our hospital with left lower abdominal pain and vaginal bleeding. A unilocular cystic tumor, measuring 13 x 10 x 10 cm, was found in her left ovary and was removed. The tumor contained clear serous fluid, approximately 600 mL, and 2 mural nodules, up to 7.5 x 5.5 x 4.5 cm. The internal cystic wall was thin for the most part and lined with ciliated cuboidal epithelium without any malignant feature. The mural part was composed of mainly more mature muscle fibers with easily discernible cross-striations, set in abundant myxoid to fibromyxoid stroma, similar to clinical and microscopic manifestations of genital rhabdomyomas reported in other sites. Because extracardiac rhabdomyoma has never been described occurring in the ovary, especially arising in serous cystadenoma, to our knowledge, this is the first case reported in the English literature. PMID- 15892007 TI - Acute hepatitis E: a cause of lymphocytic destructive cholangitis. AB - Lymphocytic destructive cholangitis is a histological pattern associating bile duct intraepithelial lymphocytic infiltration and bile duct epithelial damage. Lymphocytic destructive cholangitis is an important diagnostic feature of primary biliary cirrhosis, but it can also be seen in primary sclerosing cholangitis, autoimmune hepatitis, the so-called overlap syndrome, acute or chronic viral hepatitis C, drug-induced hepatitis, and acute rejection or graft-versus-host disease in liver or bone marrow transplantation. In the present paper we report a case of acute hepatitis with lymphocytic destructive cholangitis on liver biopsy. Clinical and biological examinations showed that the patient had hepatitis E with no other cause of liver disease. Therefore, hepatitis E should be considered as a diagnostic possibility when liver biopsy shows acute hepatitis and lymphocytic destructive cholangitis. The mechanism of bile duct damage in hepatitis E remains unknown. PMID- 15892008 TI - Well-differentiated extraskeletal osteosarcoma: report of 2 cases, 1 with dedifferentiation. AB - Extraskeletal osteosarcoma (ESOS) is a rare soft tissue sarcoma, typically characterized by high-grade histological features and a grave prognosis. However, 4 cases of well-differentiated ESOS with a better prognosis have been documented in the literature within the last 40 years. We report 2 additional cases, 1 with multicentric presentation and dedifferentiation, and we emphasize the histological features that are useful in distinguishing this lesion from other soft tissue tumors. Well-differentiated ESOS seems to represent a rare but distinct low-grade variant of ESOS. The limited published experience suggests that although the biologic behavior of this tumor is better than that of classical ESOS, there are cases with progression to a higher grade, leading eventually to final demise. PMID- 15892009 TI - Ganglioneuromatous paraganglioma of the cauda equina--a pathological case study. AB - This study presents a rare case of compound paraganglioma/ganglioneuroma with comprehensive immunohistochemical studies that reveal strong cytokeratin expression in all components. A 74-year-old woman presented with a mass lesion of the cauda equina. The 1.8-cm tumor showed 3 histomorphologically and immunohistochemically distinct components: typical paragangliomatous neuroendocrine areas, mature ganglion cell-like neuronal areas, and a "neuromatous" proliferation of Schwann cells with admixed axons. As often seen in cauda equina paragangliomas, the neuroendocrine cells were cytokeratin-positive. In addition, immunoreactivity for cytokeratins was also observed in the neurons and axons. This tumor illustrates the broad spectrum of divergent differentiation that can be seen in cells of sympathoadrenal lineage. PMID- 15892010 TI - BK virus nephropathy and vascular endothelium. PMID- 15892011 TI - Insulinlike growth factor-1 receptor: predictive factor in breast cancer patients treated with trastuzumab? PMID- 15892015 TI - The effect of intrathoracic heart position on electrocardiogram autocorrelation maps. AB - We studied the influence of the heart position in the thorax on the autocorrelation (AC) maps consisting of correlation coefficients between each pair of instantaneous electrocardiogram potential distributions over a time interval. We used a thorax-shaped electrolytic-filled tank with an isolated and perfused dog heart placed at positions spanning 5 cm on each space direction. The correlation coefficient between QRST AC maps was in the range of 0.92 to 0.99, whereas the correlation coefficient between the corresponding QRST integral maps was in the range of 0.55 to 0.87, proving that AC maps are less influenced by the heart position than integral maps. Thus, diagnostic indexes computed from the AC maps can be expected to be more specific to phenomena taking place in the myocardium than to criteria based directly on electrocardiogram amplitudes in various leads. PMID- 15892016 TI - Editorial comment. PMID- 15892017 TI - Comparison between human and automated electrocardiographic waveform measurements for calculating the Anderson-Wilkins acuteness score in patients with acute myocardial infarction. AB - The Anderson-Wilkins (AW) electrocardiographic (ECG) acuteness score complements time from pain onset in prognostic stratification of patients with acute myocardial infarction (AMI). However, for the AW acuteness score to be of practical use in the acute situation, it must be an integral component of a commercial automated ECG analysis program. The objective of this study was to determine the concordance between human and computer measurements and calculation of the AW acuteness score. The mean difference in AW acuteness score was 0.11 +/- 0.66 for anterior and -0.07 +/- 1.24 for inferior AMI. Ninety-nine percent of the differences were found to be 1.0 or less for the anterior AMI group, and 91.7% were 1.0 or less in the inferior AMI group. The differences were primarily caused by minor disagreements in measurements. In conclusion, the AW acuteness score established using manual ECG waveform measurements can be implemented into commercial automated ECG analysis programs to achieve practical use in clinical decision support for patients with AMI. PMID- 15892018 TI - Prediction of decrease in myocardial perfusion defect size and severity during a 3-month follow-up by the degree of acute resolution of electrocardiographic changes. AB - Myocardial perfusion in infarct-related artery (IRA) distribution improves progressively until a few months after successful reperfusion therapy. We assessed the rate of electrocardiographic (ECG) stage dynamics to predict perfusion improvement after mechanical, thrombolytic, or spontaneous recanalization of IRA. Thirteen patients were divided into group A (n = 8, with > or = 2 ECG stages per 2-day change rate) and group B (n = 5, no rapid change of ECG stages). There were no significant technetium Tc 99m sestamibi scintigraphic differences between the groups 3 days after recanalization; however, after 3 months, perfusion deficit size (2.8 +/- 1.8 vs 4.8 +/- 1.2, P < or = .03) and severity (1.8 +/- 0.9 vs 3.0 +/- 0, P < or = .03) were smaller in group A vs group B. The prediction sensitivity of the method was 87.5% for decrease in size and 100% for decrease in severity of perfusion defect; the specificity was 80% and 100%, respectively. A change rate of 2 or more ECG stages per 2 days predicts follow-up improvement of myocardial perfusion after IRA recanalization. PMID- 15892019 TI - Bedside risk stratification after acute myocardial infarction: prospective evaluation of the use of heart rate and left ventricular function. AB - OBJECTIVE: This study evaluated the predictive power of heart rate (HR) assessed from the standard 12-lead electrocardiogram (ECG) or from Holter recordings for future mortality and arrhythmic events in survivors of acute myocardial infarction (AMI). METHODS: Data from 432 consecutive survivors of AMI (343 men, 89 women; mean [SD] age, 58 [11] years) were analyzed. Heart rate was assessed from a standard 12-lead ECG and from 24-hour Holter recordings obtained at hospital discharge. In addition, left ventricular ejection fraction (LVEF) was noninvasively determined. The study end point was prospectively defined as a composite end point comprising mortality and arrhythmic events (ie, sudden death, resuscitated ventricular fibrillation, sustained ventricular tachycardia). Patients were followed for an average (SD) of 41 (25) months. RESULTS: Patient age, LVEF, and HR were univariate risk predictors of event-free survival. Multivariate analysis by means of a stepwise regression analysis revealed LVEF ( chi2 11.4, P = .0007), age ( chi 2 9.2, P = .02), and HR assessed from the standard 12-lead ECG ( chi2 7.1, P = .008) as independent risk parameters. CONCLUSIONS: Bedside risk stratification of survivors of AMI is feasible using simple parameters such as age, LVEF, and HR. PMID- 15892020 TI - Electrocardiogram of acute ST-elevation myocardial infarction: the significance of the various "scores". AB - The Electrocardiogram has extensively been used for evaluation and triage of patients with acute chest pain. The clinician admitting a patient with ST elevation acute myocardial infarction should be able to estimate the size and location of the ischemic area at risk, how much of the ischemic myocardium has already undergone irreversible necrosis by the time of presentation, and the "severity of ischemia" (or what is the rate of progression of necrosis as long as ischemia continues). The electrocardiographic variables that are used to make these estimates are the initial portion of the QRS (Q and R waves), the terminal portion of the QRS (the S waves and the J-point), the ST segment, and the configuration of the T waves. This editorial discuss the ability to predict each of the "physiological" parameters using the above mentioned electrocardiographic variables. PMID- 15892021 TI - Where is the central terminal located? In search of understanding the use of the Wilson central terminal for production of 9 of the standard 12 electrocardiogram leads. AB - The aim of this study was to evaluate the understanding of the term central terminal (CT) and to consider the consequences of this level of understanding. A total of 150 questionnaires was distributed during the 30th International Congress of Electrocardiology 2003, Helsinki, Finland; 42 (28%) of the anonymous questionnaires returned were considered adequate for the purpose of this study. The questionnaire addressed the following areas of interest: (1) the location of the CT; (2) the location of the negative poles of unipolar leads ; (3) the naming of the electrocardiogram lead groups; (4) the relationship between the leads and cardiac electrical views; and (5) impact on accuracy of clinical diagnosis. The findings revealed diversity in understanding the basic term, a shift in understanding the term CT to abstract/theoretical understanding, and gaps in understanding the concept of CT and the more recent theories of the cardiac electric field. PMID- 15892022 TI - Elusive understanding of electrocardiographic lead networks. PMID- 15892023 TI - Improved capacity of exercise electrocardiography in the detection of coronary artery disease by focusing on diagnostic variables during the early recovery phase. AB - The temporal distribution of the diagnostic information for the detection of coronary artery disease (CAD) provided by exercise-induced electrocardiographic (ECG) ST-segment amplitude changes in different ECG leads in men and women has not been fully investigated. To shed further light in this area, 1877 electrocardiograms selected from 8322 patients undergoing a routine exercise test on a bicycle ergometer were evaluated. ST-segment amplitude and the difference between heart rate-matched recovery and exercise ST-segment amplitudes (ST/HR difference) were measured. Coronary artery disease was verified angiographically in 669 patients and excluded in 1208 patients by angiography (n = 119), by myocardial scintigraphy (n = 250), or on clinical grounds (n = 839). The diagnostic performance of the 2 ECG methods used was assessed by constructing receiver operating characteristic curves for each sampling point every 12 seconds during 10 minutes of recovery as well as the last 4 minutes of exercise for the ST-segment amplitude. ST-segment amplitude performed better after exercise than during exercise and best within the first 2 minutes of recovery. Its diagnostic ability did not differ from the ST-amplitude hysteresis assessed by the difference between recovery ST-segment amplitude and exercise ST-segment amplitude at matched heart rate. Both methods performed better in men and the diagnostic information appeared mainly in leads I, -aVR, II, V 4 , V 5 , and V 6 . The best discrimination of CAD is provided by analysis of ST-segment amplitude changes in 6 specific leads early during the recovery phase. This information should be targeted by exercise ECG diagnostic methods. PMID- 15892024 TI - Short-term repeatability of electrocardiographic spatial T-wave axis and QT interval. AB - Although ventricular repolarization abnormalities reflect arrhythmic susceptibility, few reliable tools exist to identify their presence. We investigated the repeatability of the spatial T-wave axis and QT interval from standard 12-lead electrocardiograms in 63 asymptomatic volunteers. Certified technicians used a standardized protocol to digitally record 2 electrocardiograms per participant at each of 2 visits separated by 1 to 2 weeks. Absolute paired differences within and between visits were 0.19 degrees and 0.90 degrees for the T-wave axis and 1.08 and 1.55 milliseconds for the QT interval, respectively. The intraclass correlation coefficients for the T-wave axis and QT interval were 0.87 and 0.86, respectively. The impact of repeated measurements on the precision of the QT-interval measurements was evaluated for a hypothetical clinical trial aimed at detecting a drug-induced QT prolongation. We conclude that the spatial T wave axis is as repeatable a measure of ventricular repolarization as the QT interval. PMID- 15892025 TI - Lithium concentration correlates with QTc in patients with psychosis. AB - QT prolongation induced by antipsychotics has been reported to be a determinant for the development of torsade de pointes and sudden death. However, the effect of lithium on QT interval has not been fully clarified. The aim of the present study was to elucidate the relationship between serum lithium concentration and QT interval in patients treated with lithium. We examined serum lithium concentrations and electrocardiographic features in 39 inpatients with bipolar affective disorder or schizophrenia. The longest QT interval in the 12 electrocardiographic leads was measured using GE Marquette QT guard System Software, and Bazett formula was used for heart rate correction. The longest QTc was positively correlated with lithium concentration ( r = 0.46, P = .003). Multiple regression analysis revealed that sex (female, P = .037), lower serum K + concentration ( P = .029), and especially, higher serum lithium concentration ( P = .009) were determinants for the prolongation of the QTc. PMID- 15892026 TI - Autonomic modulation of the U wave during phenylephrine and esmolol infusions. AB - The etiology of U waves on the electrocardiogram is uncertain. We previously found that U waves could be amplified by isoproterenol infusion. Whether other autonomic interventions modify U-wave amplitude is unknown. Twenty-five healthy subjects underwent graded infusions of esmolol and phenylephrine. Heart rate, T wave amplitude, U-wave amplitude, and QT and QU intervals were measured using interactive computerized electrocardiogram software. Heart rate decreased with both interventions ( P < .001), though to a greater degree with phenylephrine. U wave amplitude increased by 16.3% during phenylephrine ( P < .001) but decreased by 14.5% during esmolol ( P < .001). The change in U-wave amplitude was strongly dose-dependent for both phenylephrine ( P < .001) and esmolol ( P < .001). T-wave amplitude increased with phenylephrine (597-692 microV, P < .001) but decreased with esmolol (632-568 microV, P < .001). QT and QU intervals increased with both interventions ( P < .001). We conclude that sympathetic and parasympathetic activities are important determinants of U-wave timing and amplitude. PMID- 15892027 TI - Wenckebach's periods or alternating Wenckebach's periods during 4:1 atrioventricular block? AB - This article deals with the tracings of a patient having episodes of atrial flutter with 4:1 atrioventricular (AV) block increasing to 8:1 AV block. This phenomenon was attributed to the coexistence of 3-level block due to transverse dissociation with 2:1 AV block in the most proximal level, or first level as well as in the second level coexisting with a Wenckebach's period in the most distal, or third level. It is suggested that perhaps the term Wenckebach's periods during progression of 4:1 AV block best describes this arrhythmia so as to maintain the term alternating Wenckebach's periods only to those occurring when 2:1 block increases to higher degrees of AV block. PMID- 15892028 TI - Apparent amelioration of bundle branch blocks and intraventricular conduction delays mediated by anasarca. AB - Nine patients aged 76.2 +/- 8.0 years, 3 males and 6 females, who had suffered anasarca due to various illnesses, and had bundle branch block (BBB) or intraventricular conduction delays (IVCD) on admission (3 with complete left BBB [LBBB], 2 with incomplete LBBB, 2 with complete right BBB, and 2 with complete IVCD), are described. After gaining 48.2 +/- 26.9 lb because of anasarca, their electrocardiograms revealed decreased QRS duration (112.7 +/- 18.9 milliseconds), as compared with the same measurement on admission (130.1 +/- 12.6 milliseconds) ( P = .015), with 4 patients showing apparent conversion of their complete BBB or IVCD to incomplete BBB, or normal IVC. Of these 9 patients, 5 lost subsequently 45.6 +/- 37.0 lb, and this was now associated with an increase in the QRS duration from 109.6 +/- 23.0 to 127.8 +/- 17.4 milliseconds ( P = .004), with 1 patient converting from normal IVC to incomplete LBBB, and 1 from incomplete to complete IVCD. These findings suggest that anasarca leads to apparent amelioration of the features of BBB or IVCD, with subsequent return to the baseline complete BBB or IVCD after loss of fluid overload; consequently, accurate characterization of a patient's BBB or IVCD is interfered with by the presence of anasarca. The pathophysiologic mechanism of this phenomenon has been traced to decreases of voltage across the entire electrocardiographic curve due to attenuation of the electrical impedance of the body volume conductor due to accumulation of the anasarca fluid. PMID- 15892029 TI - Electrocardiographic characteristics in low atrial septum pacing. AB - AIM: The aim of the study was to compare P-wave morphology and duration in pacing from the low right atrial septal wall and the high right atrial appendage (RAA). METHODS: The electrocardiogram (ECG) of 50 patients with low atrial septum (LAS) pacing and that of 50 patients with RAA pacing were compared with their electrocardiogram during sinus rhythm. RESULTS: In the frontal plane, patients with LAS pacing showed a superior P-wave axis between -60 degrees and -90 degrees . In all patients with RAA pacing, a P-wave axis between 0 degrees and +90 degrees was observed as in sinus rhythm. In the horizontal plane, all patients with LAS pacing had an anterior P-wave axis between +90 degrees and +210 degrees , whereas all patients with RAA pacing had a posterior P-wave axis between -30 degrees and -90 degrees . The terminal part of biphasic P waves in lead V 1 in LAS pacing was always positive, a pattern that was never observed in P waves of sinus origin or in RAA pacing. P-wave duration was longer with RAA pacing compared with LAS pacing (115 +/- 19 vs 80 +/- 14 milliseconds [ P < .01]). CONCLUSION: The total atrial activation time during LAS pacing is shorter than that during RAA pacing. The electrical atrial activation sequences in LAS pacing and RAA pacing are significantly different. The morphology of biphasic P waves in lead V1 during LAS pacing suggests that the initial part of activation occurs in the left atrium and the terminal part in the right atrium. PMID- 15892030 TI - Self-assembly of an interlaced triple-stranded molecular braid with an unprecedented topology through hydrogen-bonding interactions. PMID- 15892031 TI - Diffusion of water in a nonporous hydrophobic crystal. PMID- 15892032 TI - Construction of highly functionalized medium-sized rings: synthesis of hyperforin and perforatumone model systems. PMID- 15892033 TI - Irreversible inhibition of metallo-beta-lactamase (IMP-1) by 3-(3 mercaptopropionylsulfanyl)propionic acid pentafluorophenyl ester. PMID- 15892034 TI - Increasing incidence of type 1 diabetes in population from Santiago of Chile: trends in a period of 18 years (1986-2003). AB - STUDY OBJECTIVE AND SETTING: The aim of this study is to determine the incidence rate of type 1 diabetes in the metropolitan region of Santiago, Chile from 1 January 1986 to 31 December 2003. DESIGN: Population-based incidence study. PARTICIPANTS: A case must fulfill the following requirements to be included in this study: age at onset: 0 to 14 years, diagnosed with diabetes and placed on insulin, diagnosed within the defined time period, and to be a resident of the metropolitan region of Santiago at the time of the diagnosis. The population 'at risk' is the population less than 15 years of age. RESULT: The overall rate of type 1 diabetes was estimated as 4.02 cases per 100 000 children per year (95% confidence interval: 2.98-4.83). CONCLUSION: The incidence of type 1 diabetes in Santiago, Chile has increased during the last years. This data are concordant with the observation that the incidence of type 1 diabetes is increasing in Latin America and worldwide. PMID- 15892035 TI - An animal model of SARS produced by infection of Macaca mulatta with SARS coronavirus. AB - A new SARS animal model was established by inoculating SARS coronavirus (SARS CoV) into rhesus macaques (Macaca mulatta) through the nasal cavity. Pathological pulmonary changes were successively detected on days 5-60 after virus inoculation. All eight animals showed a transient fever 2-3 days after inoculation. Immunological, molecular biological, and pathological studies support the establishment of this SARS animal model. Firstly, SARS-CoV-specific IgGs were detected in the sera of macaques from 11 to 60 days after inoculation. Secondly, SARS-CoV RNA could be detected in pharyngeal swab samples using nested RT-PCR in all infected animals from 5 days after virus inoculation. Finally, histopathological changes of interstitial pneumonia were found in the lungs during the 60 days after viral inoculation: these changes were less marked at later time points, indicating that an active healing process together with resolution of an acute inflammatory response was taking place in these animals. This animal model should provide insight into the mechanisms of SARS-CoV-related pulmonary disease and greatly facilitate the development of vaccines and therapeutics against SARS. PMID- 15892036 TI - Trophic transfer of cyanobacterial toxins from zooplankton to planktivores: consequences for pike larvae and mysid shrimps. AB - The aim of this study was to evaluate the potentially harmful effects of zooplankton preexposed to cyanobacteria on two planktivorous animals: a fish larva (pike, Esox lucius) and a mysid shrimp (Neomysis integer). The planktivores were fed zooplankton from a natural community that had been preexposed to cell free extract or to purified toxin (nodularin) of the cyanobacterium Nodularia spumigena, and the growth, feeding, and pellet production of the planktivores, as well as the toxin content of the pellets, were measured. In addition, radiolabeled nodularin ((3)H-dihydronodularin) was used in separate experiments to measure the vector transfer of nodularin from zooplankton to their predators. During 11-day exposures, dissolved nodularin was transferred to pike larvae and N. integer via zooplankton at very low rates of accumulation. Treatment with N. spumigena extract decreased the ingestion and feces production rates of pike larvae. With purified nodularin alone, no such effect could be observed. No effect on molting cycle length, fecal pellet production, C:N ratio, or growth of N. integer was detected. The results suggest that dissolved cyanobacterial toxins released during bloom decay can have a negative impact on feeding and, hence, on the growth of fish larvae via zooplankton, even without direct contact between cyanobacteria and the fish. PMID- 15892037 TI - Insecticidal compounds of the biofilm-forming cyanobacterium Fischerella sp. (ATCC 43239). AB - Cyanobacterial biofilms are grazed by many different benthic invertebrates. In particular, larvae of insects are often present in great numbers and exert strong grazing pressure on cyanobacteria. Along with morphological adaptations, allelochemicals may have been developed as defense mechanisms against insect larvae. To investigate the chemical defense of biofilm-forming cyanobacteria, larvae of Chironomus sp., a widely distributed genus in this habitat, were used. Ten artificial biofilms of axenic and nonaxenic cyanobacteria were screened for insecticidal activity against Chironomus sp. Fischerella 43239 was the cyanobacterium that exhibited the highest acute toxicity. A bioassay-guided isolation procedure was used to study the compounds responsible for toxicity in more detail. A toxic fraction was obtained when the 60% methanolic extract of Fischerella 43239 was separated on a C18 HPLC column. Electrospray mass spectrometry indicated the presence of several compounds in this fraction. The successful separation into individual compounds was achieved by HPLC on a cyanopropyl column. The heavily clustered quasimolecular ions observed on an electrospray mass spectrometer and the absorption spectra of the separated compounds were indicative of indole derivatives. The existence in the benthic cyanobacterium Fischerella 43239 of strong insecticidal metabolites that serve as chemical protection agents against insect larvae is supported by the data. PMID- 15892038 TI - Effects of microcystins on and toxin degradation by Poterioochromonas sp. AB - A Chrysophyceae species, Poterioochromonas sp., was isolated from Microcystis cultures. This species can efficiently prey on Microcystis and can grow faster phagotrophically than autotrophically. The growth of Poterioochromonas sp. was stimulated in the presence of microcystin-LR and microcystin-RR (in concentrations ranging from 0.1 to 4 mg/L). The growth rate of Poterioochromonas was 4-5 times higher than the control, indicating the toxins serve as growth stimuli for this organism. A subculture of toxin-treated cells, however, showed low cellular viability, suggesting that growth enhancement by microcystins was not a normal process. The antioxidant enzymatic activity of Poterioochromonas sp. was screened for toxicology analysis. Glutathione, malondialdehyde, and superoxide dismutase (SOD) content was up-regulated within 8 h of exposure to microcystin-LR (500 microg/L). A high level of SOD activity during exposure to the toxin indicated that SOD was involved in decreasing oxidative stress caused by microcystin-LR. Simultaneously with growth, Poterioochromonas was able to degrade microcystin-LR even, at a toxin concentration of 4 mg/L. This putative degradation mechanism in Poterioochromonas is explored further and discussed in this article. Our findings may shed light on understanding the role of Poterioochromonas in the aquatic ecosystem, in particular, as a grazer of toxic cyanobacteria and a biodegrader for microcystins. PMID- 15892039 TI - Quantitative LC-ESI-MS analyses of microcystins and nodularin-R in animal tissue- matrix effects and method validation. AB - The matrix effects and signal response in LC-MS analysis of six microcystins and nodularin-R were studied in mussels and liver samples from the common eider and rainbow trout. The instrumentation used in the study was a triple quadrupole MS with electrospray ionization. The results from the spiked tissue samples showed that both signal suppression and enhancement occurred. The recorded matrix effects were not severe; all studied toxins could be detected with sufficient limit of detection in all matrices. The results indicate, however, that matrix effects must be monitored for accurate quantification of microcystin and nodularin in tissue samples. Matrix effects can be studied with standard additions in the studied matrix, as was done in this study. Solid-phase extraction (SPE) resulted in a lower limit of detection compared to no cleanup in the sample preparation. SPE also prolonged the chromatographic stability. SPE cleanup is therefore strongly recommeded. Also described in this article are the chromatographic and mass spectrometric details of glutathione and cysteine conjugates, which are the detoxification products of the toxins. LC-MS analysis is suitable for detoxification studies of microcystins and nodularins. Cysteine conjugate was identified as the main detoxification product in a mussel sample that was exposed to toxic cyanobacteria in an aquarium. PMID- 15892040 TI - Bovine mater-like NALP9 is an oocyte marker gene. AB - We have cloned and sequenced the bovine NALP9 cDNA, which contains a 2991 bp long open reading frame with 76% homology to its human counterpart. The putative 996 amino acids protein presents the domain organization typical of the Nacht, Leucine rich repeat and Pyrin domain containing (NALP) family that includes the maternal effect factor MATER. By reverse transcription coupled to polymerase chain reaction, we have analyzed expression of NALP9 in a panel of somatic and gonadic tissues and during in vitro preimplantation development. The transcript was detected exclusively in testis and ovary, and at a remarkably high level in the oocyte from antral follicles. NALP9 mRNA remained present after in vitro maturation and fertilization, and was detected in embryos, but transcription was not reactivated at the maternal to embryo transition. Thus NALP9 is characterized as a novel oocyte marker gene in cattle. PMID- 15892041 TI - Pressurized liquid extraction of toxins from cyanobacterial cells. AB - The suitability of pressurized liquid extraction (PLE) of cyanotoxins from cells was investigated. The stability of cyanotoxins (MCYST-RR, MCYST-LR, and anatoxin a) was evaluated at nine combinations of pressure and temperature (7, 10, and 14 MPa and 60 degrees C, 80 degrees C and 100 degrees C) using 75% (v/v) methanol in water (MeOH) as solvent. Additional experiments investigated the stability of cyanotoxins when water was used as solvent (at a pressure of 14 MPa and a temperature of 40 degrees C, 50 degrees C, 60 degrees C, 80 degrees C, or 100 degrees C). Results using 75% MeOH showed that the MCYST-RR and MCYST-LR were stable under the tested pressures up to 80 degrees C. At 100 degrees C MCYST recovery decreased by 10% to 17%. When water was used as the solvent, no differences in recovery were observed for MCYST-LR, whereas for MCYST-RR, maximum recovery was obtained at 60 degrees C, and degradation occurred at 100 degrees C. In contrast, anatoxin-a was labile under all experimental conditions; the best recoveries (ca. 50%) were obtained at 60 degrees C at the three pressures using 75% MeOH. However, only 17%-23% recovery was obtained with water extraction at all temperatures. The extraction of MCYST-LR and variants from cells (Microcystis aeruginosa, UTCC299) was studied using two solvents, 75% MeOH and 100% water, at 14 MPa and 60 degrees C and 100 degrees C. PLE extracts were compared with extracts obtained with 75% MeOH and ultrasonication. Complete extraction was achieved in both solvents in one 5-min cycle (at 100 degrees C). Although lower recovery was obtained using PLE (79%-105%), shorter extraction time and automation are advantageous over ultrasonication. PMID- 15892042 TI - Full-term development of mouse eggs transplanted with male pronuclei derived from round spermatids: the effect of synchronization between male and female pronucleus on embryonic development. AB - Pronucleus transplanted mice have been produced, but their donor male pronuclei were derived from mature sperm and were completely synchronous with female pronuclei because both male and female pronuclei came from the same fertilized oocyte. The present study firstly produced male pronuclei by introducing round spermatids into enucleated mouse oocytes, then transferred the male pronuclei into mouse oocytes at three activation stages and finally compared the effect of three kinds of oocytes on the development of reconstructed embryos. Our results indicate that, in enucleated oocytes, mouse round spermatid nuclei can transform to male pronuclei in a higher proportion, and the synchronization between male and female pronucleus does not significantly influence the early cleavage but the later and full-term development of reconstructed embryos. PMID- 15892043 TI - Capecitabine plus docetaxel combination therapy. AB - BACKGROUND: For patients with anthracycline-pretreated metastatic breast carcinoma, capecitabine plus docetaxel significantly increased overall survival compared with docetaxel alone. The current study evaluated the cost-effectiveness of the capecitabine/docetaxel combination versus docetaxel monotherapy, comparing the gain in quality-adjusted survival with associated health care costs. METHODS: Patients were randomized to receive 21-day cycles of oral capecitabine 1250 mg/m2 twice daily, on Days 1-14, plus docetaxel 75 mg/m2 Day 1 (n = 255), or docetaxel 100 mg/m2 on Day 1 (n = 256). Health and cost outcomes in the two arms were compared, and cost-effectiveness was estimated. Data on survival time and medical care resource use were prospectively collected in the trial. Costs associated with medical care resource use and quality-of-life adjustments were obtained from the published literature. The incremental cost-effectiveness ratio was calculated as the cost per quality-adjusted life year (QALY) gained. RESULTS: Capecitabine/docetaxel increased the median overall survival by 3 months compared with docetaxel alone (14.5 vs. 11.5 months). The mean quality-adjusted survival was increased by 1.8 months in the capecitabine/docetaxel group. The total medical-resource utilization cost per patient was 8.9% higher with the combination: 24,475 dollars for combination therapy versus 22,477 dollars for single-agent docetaxel. The mean cost per QALY gained with combination therapy was 13,558 dollars (standard deviation, 6742 dollars). Cost savings due to reduced docetaxel dose and hospital use were the major cost offsets with the combination. Sensitivity analyses showed that varying the mean hospital cost per day from the 5th to the 95th percentile resulted in cost-utility ratios ranging from 20,326 dollars to as low as 6360 dollars. CONCLUSIONS: Capecitabine/docetaxel was a cost-effective treatment in patients with anthracycline-pretreated advanced breast carcinoma, and had an incremental cost effectiveness ratio that compares very favorably with that of many other oncology therapies. PMID- 15892044 TI - Functional challenge affects aquaporin mRNA abundance in mouse blastocysts. AB - The aquaporins (AQPs) are a family of channel proteins that facilitate diffusion of water across cell membranes. Three members of the AQP family have been detected in the mouse blastocyst: AQP 3 and 8 are located in the basolateral domain and AQP 9 predominantly in the apical domain of the trophoblast cells. These are believed to be involved in facilitating the accumulation of fluid into the blastocyst cavity. We have investigated the ability of mouse embryos to regulate AQP gene expression in response to different treatments expected to affect the passage of water across the trophoblast cells using real-time PCR. In the first experiment 8-cell embryos were allowed to develop to blastocysts in media from 300 to 400 mOsm. Blastocyst formation was unaffected by media made hyperosmolar by glycerol, whereas blastocyst formation was significantly reduced in sucrose-based 350 and 400 mOsm media. AQP 8 mRNA levels were reduced when embryos were cultured in glycerol-based hyperosmolar media. The mRNA levels of AQP 3, 7, 9, and 11 were not significantly affected by hyperosmolar media. In the second experiment blastocysts were punctured (0 hr) and allowed to re-expand. AQP mRNA levels were examined after 2, 6, and 10 hr. Compared to control embryos, the expression of AQP 3, 7, and 9 were upregulated after 2 hr. Upregulation was sustained only for AQP 9 and this was sustained up to 6 and 10 hr after puncture. In the third experiment we compared expression of AQPs between in vitro cultured and in vivo developed blastocysts. We found that in vitro culture resulted in lower levels of AQP 8, 9, and 11 compared to in vivo development. These experiments show that mouse embryos are capable of regulating AQP mRNA abundances in response to environmental alterations. PMID- 15892045 TI - Identification of the bull sperm p80 protein as a PH-20 ortholog and its modification during the epididymal transit. AB - We have identified an 80 kDa protein in ejaculated bull spermatozoa (p80) which is found in acrosomal and post-acrosomal areas of the head. It has a hyaluronidase activity and shares homologies with PH-20, a sperm surface glycoprotein involved in sperm-egg interaction. The aim of the present study was to characterize bull sperm p80 protein at the nucleic and amino acid levels to determine whether it is the bovine PH-20 ortholog. The complete nucleotide sequence determined by RT-PCR, 3' and 5' RACE show that bull p80, displays identity with the PH-20 nucleotide and amino acid sequences. Messenger RNA and protein expressions determined by Northern blot and immunohistochemistry revealed that the protein is testicular (expressed in spermatocytes and spermatids). The localization of p80 on spermatozoa, determined by indirect immunofluorescence using a monoclonal antibody, shows the protein in acrosomal and post acrosomal areas of the head with an increase in the signal intensity as sperm progress through the epididymis. Post-translational modifications of the protein were investigated during the epididymal maturation by Western blot on protein extracts from sperm collected in the caput, corpus and cauda portions of bull epididymis. Glycolysation status of sperm p80 protein on proteins from ejaculated and epididymidal sperm was investigated. Result show that the glycosylation status is modified as spermatozoa migrate through the epididymis. Hyaluronidase activity evaluated in protein extracts from spermatozoa of the three different epididymal sections revealed that the activity is higher at pH 7 than 4 and is not affected by epididymal maturation. These data strongly suggest that p80 is the bovine PH 20. PMID- 15892046 TI - Isolation and characterization of the major proteins of ram seminal plasma. AB - Mammalian seminal plasma contains among others, two major families of proteins, namely spermadhesins and those proteins that contain fibronectin type II domains. Spermadhesins are the major proteins of boar and stallion seminal plasma and homologous proteins have been identified in the bull. These proteins appear to be involved in capacitation and sperm-egg interaction. In bovine seminal plasma, proteins containing fibronectin type II domains are the major proteins and are designated BSP proteins. These proteins play a role in sperm capacitation. In this study, we present the isolation and characterization of the major proteins of ram seminal plasma. Precipitated proteins from Suffolk ram seminal plasma were loaded onto a gelatin-Agarose column. The unadsorbed (fraction A) and retarded proteins (fraction B) were removed by washing the column with phosphate buffered saline and the adsorbed proteins (fraction C) were eluted with 5 M urea. SDS-PAGE of fraction B indicated the presence of a 15.5 kDa protein, which is the major protein of ram seminal plasma (approximately 45% of total protein by weight) and was identified as a spermadhesin by N-terminal sequencing. SDS-PAGE analysis of fraction C revealed the presence of four proteins, which represented approximately 20% of total ram seminal plasma proteins by weight, and were identified as proteins of the BSP family and named RSP proteins. These RSP proteins were designated RSP-15 kDa, RSP-16 kDa, RSP-22 kDa, and RSP-24 kDa. Only RSP-15 kDa and -16 kDa proteins cross-reacted with antibodies against BSP proteins. Ram spermadhesin and RSP proteins interact with heparin but only RSP proteins bind to hen's egg yolk low-density lipoprotein. In conclusion, spermadhesin is the major protein of ram seminal plasma and other major proteins belong to the BSP protein family. PMID- 15892048 TI - Cyanobacteria/cyanotoxin research--looking back for the future: the opening lecture of the 6th ICTC, Bergen, Norway. AB - A retrospective view of the development of research on cyanobacteria and their toxins-in Norway and worldwide-is cursorily examined. Combined with a personal narrative of the work and life of the author, the relevant research progress in the last 50 years of the 20th century is outlined. This was the period in biology when research of toxigenic cyanobacteria was coming to a head. The knowledge and understanding achieved were the result of a collective international enterprise binding the community of specialists together. The noxious organisms were isolated, and the basic chemistry, physiology, and toxicology of the specific cyanotoxins were settled. At the threshold today of what is widely regarded as the century of biology, the study of toxigenic cyanobacteria is undergoing important transformations. New frontiers include the interface between molecular biology, biochemistry, toxicology, and ecology. The results of the basic and applied research efforts will expand the understanding of cyanobacteria and be beneficial to the management of natural resources and the care of human beings. The tiny cyanobacteria, being a geological force in the dynamics of the globe, will continually become more central to the modern world and be applied more closely, affecting our lives. PMID- 15892049 TI - Germ-cell specific protein gametogenetin protein 2 (GGN2), expression in the testis, and association with intracellular membrane. AB - Gametogenetin (Ggn) is a germ cell-specific gene with multiple splicing variants giving rise to three predicted protein products, gametogenetin protein 1 (GGN1), gametogenetin protein 2 (GGN2), and gametogenetin protein 3 (GGN3). GGN1 and GGN3 were reported to interact with Fanconi anemia complementation group L (FANCL) per proliferation of germ cells (POG), a ubiquitin E3 ligase involved in germ-cell deficient (gcd) mutation. While GGN2, another protein from Ggn by alternative splicing did not interact with FANCL/POG since it lacked the domain mediating the interaction. Little is known about the expression and function of GGN2. Here through Northern blotting experiment we showed that Ggn was mainly expressed in the testis but hardly detectable in the ovary or the somatic tissues. By preparing GGN2-specific antibody we showed that GGN2 was detectable and only detectable in the testis. By comparing the expression of Ggn mRNA and GGN2 protein in developing mouse testis, we showed that there was no evident delay of the translation of Ggn mRNA after their transcription. Both the subcellular localization study and the germ cell membrane protein fractionation implied that GGN2 associated with the intracellular membrane system. Co-fractionation on Superdex and yeast two-hybrids suggested that like GGN1, GGN2 was also a potential interaction partner of gametogenetin binding protein 1 (GGNBP1). Our data suggested that gametogenetin proteins were mainly involved in male germ cell development and GGN2 was also a possible interaction partner of GGNBP1. Like GGN1, GGN2 was also possibly involved in cell trafficking. The possible involvement of GGN2 in acrosome biogenesis was proposed. PMID- 15892050 TI - DkkL1 (Soggy), a Dickkopf family member, localizes to the acrosome during mammalian spermatogenesis. AB - Dickkopf-like 1 (DkkL1) is related to the Dickkopf gene family, a group of proteins that are characterized as secreted antagonists of Wingless (Wnt) signal transduction proteins. DkkL1 mRNA is found in preimplantation mouse embryos and in developing neural tissue, but in adults it is found primarily in the testes. In an effort to elucidate its function, the distribution of DkkL1 protein in mouse testis and mature sperm was analyzed by immuno-histochemistry and immuno blotting techniques. DkkL1 first appeared in the developing spermatocytes in seminiferous tubules as early as Stage XII, coincident with the appearance of DkkL1 mRNA. Surprisingly, however, DkkL1 localized to the developing acrosome in spermatocytes and spermatids and to the acrosome in mature sperm. Furthermore, DkkL1 was N-glycosylated in the testis, but it did not appear to be excreted, and the DkkL1 in mature sperm was no longer N-glycosylated, suggesting that additional post-translational modifications occurred during the final stages of spermatogenesis. These results identify a member of the Dickkopf family as a novel acrosomal protein that may be involved in acrosome assembly or function, a unique role for a secreted signaling molecule. PMID- 15892051 TI - Detoxification of organophosphate nerve agents by immobilized dual functional biocatalysts in a cellulose hollow fiber bioreactor. AB - A whole-cell technology for detoxification of organophosphates based on genetically engineered Escherichia coli cell expressing both cellulose-binding domain (CBD) and organophosphorus hydrolase (OPH) onto cell surface was reported recently (Wang et al., 2002). This study reports the application of these biocatalysts when immobilized in a cellulose hollow fiber bioreactor (HFB) for the biodetoxification of a model organophosphate, paraoxon, in a continuous flow mode. In 24 h, 0.79 mg wet cell/cm2 fiber surface were immobilized onto cellulose fibers specifically and strongly through the cellulose binding domain, forming a monolayer demonstrated by Scanning Electronic Micrograph, and essentially no cell was washed away by washing buffer. The immobilized biocatalyst had a high performance of detoxifying paraoxon solution of 5,220 mumol/h x L reactor or 990 mumol/h x m2 reactor. The immobilized biocatalysts maintained a stable degradation capacity for 15 uses over a period of 48 days with only 10% decline in degradation efficiency under operating and storage conditions. In addition, the bioreactor was easily regenerated by washing with 1% sodium dodecyl sulfate (SDS), with 86.7% immobilization capacity and 93.9% degradation efficiency recovery. This is the first report using the HFB in a non-traditional way, immobilizing whole-cell biocatalysts by specific adhesion thus rendering the catalysis operation the advantages of low pressure drop, low shear force, and low energy requirement. The successful application of this genetically engineered dual functional E. coli strain in a model bioreactor shows its promise in large scale detoxification of organophosphate nerve agents in bulk liquid phase. PMID- 15892052 TI - Comparison of acetate and propionate uptake by polyphosphate accumulating organisms and glycogen accumulating organisms. AB - Enhanced biological phosphorus removal (EBPR) performance is directly affected by the competition between polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigates the effects of carbon source on PAO and GAO metabolism. Enriched PAO and GAO cultures were tested with the two most commonly found volatile fatty acids (VFAs) in wastewater systems, acetate and propionate. Four sequencing batch reactors (SBRs) were operated under similar conditions and influent compositions with either acetate or propionate as the sole carbon source. The stimulus for selection of the PAO and GAO phenotypes was provided only through variation of the phosphorus concentration in the feed. The abundance of PAOs and GAOs was quantified using fluorescence in situ hybridisation (FISH). In the acetate fed PAO and GAO reactors, "Candidatus Accumulibacter phosphatis" (a known PAO) and "Candidatus Competibacter phosphatis" (a known GAO) were present in abundance. A novel GAO, likely belonging to the group of Alphaproteobacteria, was found to dominate the propionate fed GAO reactor. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to take up acetate and propionate. PAOs enriched with acetate as the sole carbon source were immediately able to take up propionate, likely at a similar rate as acetate. However, an enrichment of GAOs with acetate as the sole carbon source took up propionate at a much slower rate (only about 5% of the rate of acetate uptake on a COD basis) during a short-term switch in carbon source. A GAO enrichment with propionate as the sole carbon source took up acetate at a rate that was less than half of the propionate uptake rate on a COD basis. These results, along with literature reports showing that PAOs fed with propionate (also dominated by Accumulibacter) can immediately switch to acetate, suggesting that PAOs are more adaptable to changes in carbon source as compared to GAOs. This study suggests that the PAO and GAO competition could be influenced in favour of PAOs through the provision of propionate in the feed or even by regularly switching the dominant VFA species in the wastewater. Further study is necessary in order to provide greater support for these hypotheses. PMID- 15892053 TI - Short-term and long-term effects of increasing temperatures on the stability and the production of volatile sulfur compounds in full-scale thermophilic anaerobic digesters. AB - This study compares the effect of a rapid increase of the digester temperature (from 54 degrees C to 58 degrees C in 2 weeks) with a slow increase (from 53.9 degrees C to 57.2 degrees C at a rate of 0.55 degrees C per month) on full-scale thermophilic anaerobic digestion at Hyperion Treatment Plant. The short-term test demonstrated that rapidly increasing the digester temperature caused elevated production of volatile sulfur compounds, most notably methyl mercaptan, but volatile solids destruction and methane production were not significantly affected. The increase of the volatile fatty acid to alkalinity ratio from 0.1 to over 0.3 indicated a transient change in digester biochemistry, which was reversed by lowering the temperature. In the long term-test, a slow increase of digester temperature, the production of hydrogen sulfide increased above temperatures of 56.1 degrees C, but was controlled by increased injection of ferrous chloride. Methyl mercaptan was detected in trace amounts at the highest temperature tested (57.2 degrees C). This test showed insignificant effects on other digestion parameters, although some temperature-independent changes were observed that could have been seasonal effects over the year that the long-term test lasted. Thus a slow temperature increase was preferable. This observation contrasts with previous results showing the desirability of a rapid temperature rise to first establish a thermophilic culture when converting from mesophilic operation. Further research is warranted on temperature limits and process changes to optimize thermophilic anaerobic digestion. PMID- 15892054 TI - A simple method to estimate the contribution of biological floc and reactor solution to mass transfer of oxygen in activated sludge processes. AB - In this study, the mass transfer coefficient of biological floc (K(L)a(bf)) was estimated from the mass transfer coefficient of the mixed-liquor (K(L)a(f)) and the reactor-solution (K(L)a(e)). The biological floc resistance (BFR) and reactor solution resistance (SR) were defined as the reciprocal of K(L)a(bf) and K(L)a(e), respectively, by applying the concept of serial-resistance originally presented in two-film theory (Lewis and Whitman (1924) Ind Eng Chem 16:1215 1220). The specific biological floc resistance (SBFR) was defined as biological floc resistance per unit biomass concentration. The data indicated that an activated sludge process yielding low BFR/MLR and BFR/SR tended to produce higher oxygen transfer efficiency. Surprisingly, the reactor-solution posed the same level of resistance as clean water in all experiments, except in a 5-day SRT, non nitrifying, completely mixed activated sludge (CMAS) process run. Furthermore, SBFR successfully represented biological floc and showed a positive correlation to sludge volume index (SVI). In addition, SBFR/SR and oxygen transfer efficiency (OTE(f)) followed an exponential relationship for the complete data set. The method of separating the mixed-liquor into biological floc and reactor-solution improved the understanding of oxygen transfer under process conditions, without resorting to intrusive techniques or direct handling of fragile biological floc. PMID- 15892055 TI - (R)-phenylacetylcarbinol production in aqueous/organic two-phase systems using partially purified pyruvate decarboxylase from Candida utilis. AB - Aqueous/organic two-phase systems have been evaluated for enhanced production of (R)-phenylacetylcarbinol (PAC) from pyruvate and benzaldehyde using partially purified pyruvate decarboxylase (PDC) from Candida utilis. In a solvent screen, octanol was identified as the most suitable solvent for PAC production in the two phase system in comparison to butanol, pentanol, nonanol, hexane, heptane, octane, nonane, dodecane, methylcyclohexane, methyl tert butyl ether, and toluene. The high partitioning coefficient of the toxic substrate benzaldehyde in octanol allowed delivery of large amounts of benzaldehyde into the aqueous phase at a concentration less than 50 mM. PDC catalyzed the biotransformation of benzaldehyde and pyruvate to PAC in the aqueous phase, and continuous extraction of PAC and byproducts acetoin and acetaldehyde into the octanol phase further minimized enzyme inactivation, and inhibition due to acetaldehyde. For the rapidly stirred two-phase system with a 1:1 phase ratio and 8.5 U/mL carboligase activity, 937 mM (141 g/L) PAC was produced in the octanol phase in 49 h with an additional 127 mM (19 g/L) in the aqueous phase. Similar concentrations of PAC could be produced in the slowly stirred phase separated system at this enzyme level, although at a much slower rate. However at lower enzyme concentration very high specific PAC production (128 mg PAC/U carboligase at 0.9 U/mL) was achieved in the phase separated system, while still reaching final PAC levels of 102 g/L in octanol and 13 g/L in the aqueous phase. By comparison with previously published data by our group for a benzaldehyde emulsion system without octanol (50 g/L PAC, 6 mg PAC/U carboligase), significantly higher PAC concentrations and specific PAC production can be achieved in an octanol/aqueous two-phase system. PMID- 15892056 TI - Regulation of cell adhesion using a signal-responsive membrane substrate. AB - We have developed a novel cell culture material that regulates cell adhesion by changes in potassium ion concentration. The material is a polyethylene substrate grafted to a copolymer of the thermoresponsive polymer N-isopropylacrylamide (NIPAM) and benzo-18-crown- 6-acrylamide (BCAm), with a pendant crown ether as sensor. The crown ether recognizes potassium ion concentrations and NIPAM conformational changes lead to changes in the hydrophobicity/hydrophilicity balance of the entire polymer at constant cell culture temperatures. Although cells were successfully cultured on the ion recognition material in normal culture medium at 37 degrees C, the cells could be detached from the material surface by adding potassium ions alone, without proteolytic enzymes, because the surface to which the cells were attached altered its surface characteristics to a more hydrophilic state. Therefore, cell layers with intact cell-to-cell junctions and high activities were successfully recovered. Furthermore, by changing the target sensors, this material will be able to control cell adhesion through various cellular signals. PMID- 15892057 TI - Use of microelectrodes to investigate the effects of 2-chlorophenol on microbial activities in biofilms. AB - In order to assess the applicability of using microelectrodes as a tool for inhibition tests, temporal and spatial inhibitory effects of 2-chlorophenol (2 CP) on O(2) respiration and nitrification activities in municipal wastewater biofilms were investigated using microelectrodes for O(2) and NH(4)(+). The time course microelectrode measurements demonstrated that 2-CP inhibited O(2) respiration and nitrification activities within 6-18 min. The microbial activities were inhibited only in the upper 400 microm of the biofilms by 2-CP, and the bacteria present in the deeper parts of the biofilms were still active, probably due to limited penetration of 2-CP. These results could reasonably explain the difference in inhibitory ratios of the O(2) respiration and nitrification activities in the biofilms. O(2) respiration activity was incompletely inhibited, which was attributed to the presence of O(2) respiration activities in the deeper parts of the biofilm. In contrast, nitrification activity was significantly inhibited because ammonia-oxidizing bacteria were present in the upper parts of the biofilm. These results indicate that the microelectrodes with a very quick response time and a high spatial resolution are useful tools to study temporal and spatial inhibitory effects of inhibitors on in situ microbial activities in biofilms. PMID- 15892058 TI - Genotoxic potential of Microcystin-LR and nodularin in vitro in primary cultured rat hepatocytes and in vivo in rat liver. AB - Microcystin-LR (MCYST-LR) and nodularin (NOD) are known as tumor promoters in experimental animals and so present potential health threats for humans. Although their hepatotoxic mechanisms have been very well documented, many other effects of these toxins are relatively undescribed, indeed controversial, notably those related to their genotoxicity. In the present investigation, we examined how these toxins could induce DNA damage using a combination of in vitro and in vivo approaches. We first used the (32)P-postlabeling assay to test hydrophobic adduct formation on DNA from primary cultured rat hepatocytes treated with noncytotoxic concentrations of MCYST-LR and NOD (2 and 10 ng/mL). Analysis of the autoradiograms of DNA digests isolated from the hepatocytes did not show any hydrophobic DNA adduct formation. However, these toxins significantly decreased the amount of hydrophobic endogenous adducts, termed I compounds. We next investigated oxidative DNA damage by using the (32)P-postlabeling assay to analyze 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) content as a biomarker of possible DNA lesions. Both MCYST-LR and NOD significantly enhanced 8-oxo-dG in time- and dose-dependent manner in vitro in primary cultured hepatocytes and in vivo in rat liver cells. Thus, it appears that the depletion of endogenous DNA adducts (I compounds) and/or the increase of 8-oxo-dG levels by MCYST-LR and NOD could be involved in the formation of hepatic tumors during long-term exposure to these cyanobacterial hepatotoxins. PMID- 15892059 TI - Avian vacuolar myelinopathy linked to exotic aquatic plants and a novel cyanobacterial species. AB - Invasions of exotic species have created environmental havoc through competition and displacement of native plants and animals. The introduction of hydrilla (Hydrilla verticillata) into the United States in the 1960s has been detrimental to navigation, power generation, water intake, and water quality (McCann et al., 1996). Our field surveys and feeding studies have now implicated exotic hydrilla and associated epiphytic cyanobacterial species as a link to avian vacuolar myelinopathy (AVM), an emerging avian disease affecting herbivorous waterbirds and their avian predators. AVM, first reported in 1994, has caused the death of at least 100 bald eagles (Haliaeetus leucocephalus) and thousands of American coots (Fulica americana) at 11 sites from Texas to North Carolina (Thomas et al., 1998; Rocke et al., 2002). Our working hypothesis is that the agent of this disease is an uncharacterized neurotoxin produced by a novel cyanobacterial epiphyte of the order Stigonematales. This undescribed species covers up to 95% of the surface area of leaves in reservoirs where bird deaths have occurred from the disease. In addition, this species is rare or not found on hydrilla collected at sites where AVM disease has not been diagnosed. Laboratory feeding trials and a sentinel bird study using naturally occurring blooms of cyanobacteria on hydrilla leaves and farm-raised mallard ducks (Anas platyrhynchos) induced the disease experimentally. Since 1994 AVM has been diagnosed in additional sites from Texas to North Carolina. Specific site characteristics that produce the disjunct distribution of AVM are unknown, but it is probable that the incidence of this disease will increase with the introduction of hydrilla and associated cyanobacterial species into additional ponds, lakes, and reservoirs. PMID- 15892060 TI - Chronic toxicity and responses of several important enzymes in Daphnia magna on exposure to sublethal microcystin-LR. AB - In the current study, the toxicological mechanisms of microcystin-LR and its disadvantageous effects on Daphnia magna were examined. Survival rate, number of newborn, activity of several important enzymes [glutathione S-transferase (GST), lactate dehydrogenase (LDH), phosphatases, and glutathione], accumulated microcystins, and ultrastructural changes in different organs of Daphnia were monitored over the course of 21-day chronic tests. The results indicated that low concentrations of dissolved microcystin had no harmful effect on Daphnia. On the contrary, stimulatory effects were detected. In the presence of toxin at high dosage and for long-term exposure, GST and glutathione levels decreased significantly. The decreased enzyme activity in the antioxidant system probably was caused by detoxification reactions with toxins. And these processes of detoxification at the beginning of chronic tests may enable phosphatases in Daphnia magna to withstand inhibition by the toxins. At the same time, we also found that the LDH activity in test animals increased with exposure to microcystin-LR, indicating that adverse effects occurred in Daphnia. With microcystin given at a higher dosage or for a longer exposure, the effect on Daphnia magna was fatal. In the meantime, microcystin began to accumulate in Daphnia magna, and phosphatase activity started to be inhibited. From the ultrastructure results of cells in D. magna, we obtained new information: the alimentary canal may be the target organ affected by exposure of microcystins to D. magna. The results of the current study also suggested that the oxidative damage and PPI (protein phosphatase inhibition) mechanisms of vertebrates also are adapted to Daphnia. PMID- 15892062 TI - Microcystin-RR uptake and its effects on the growth of submerged macrophyte Vallisneria natans (lour.) hara. AB - Microcystins are hepatotoxins produced by many species of several cyanobacterial genera. Their toxic effects on animals and some terrestrial higher plants have been well studied, but their potential effects on the development of aquatic plant seedlings are not well known, and their uptake by aquatic plants is seldom reported. In our research the seeds and seedlings of the submerged macrophyte Vallisneria natans were exposed to different concentrations of microcystin-RR, which was purified with high-performance liquid chromatography (HPLC). The results indicated that microcystin-RR could accumulate differentially in the roots and leaves of V. natans seedlings. Toxin accumulation in the roots and leaves was time- and dose-dependent, with higher uptake detected in the roots. Growth and development detection revealed that V. natans was relatively insensitive to microcystin-RR at concentrations ranging from 0.0001 to 0.01 mg/L. However, when the toxin concentration was more than 0.01 mg/L, both the fresh weight and the longest leaf length of seedlings were significantly reduced after a 30-day treatment. The root and leaf numbers were significantly decreased when 10 mg/L of toxin was used. These results suggest that microcystin-RR can be taken up by V. natans, which subsequently will retard its development. PMID- 15892061 TI - First report of saxitoxin in Finnish lakes and possible associated effects on human health. AB - This study is the first report of saxitoxin in cyanobacterial blooms in Finland. Bloom samples (n = 50) were collected from Finnish freshwater sites during summer months of 2002 and 2003. These samples were screened for the presence of paralytic shellfish toxins (PSTs) using the Jellett rapid PSP screening test. Samples testing positive for PSTs (n = 7) were further analyzed with saxiphilin- and voltage-gated sodium channel [(3)H]-STX-binding radioreceptor assays and liquid chromatography using fluorescence and mass spectrometric analysis. The results indicated that saxitoxin (STX) was the only PST analogue in the samples and that it was present in high concentrations, as much as 1 mg L(-1). Microscopic analysis revealed that 95%-100% of the phytoplankton in the positive samples consisted of Anabaena lemmermannii. The trophic status of lakes in which STX-containing blooms were found varied from oligotrophic to hypertrophic. All the lakes had high nitrogen-to-phosphorus ratios. In some instances, samples had been collected from sites where swimmers had reported adverse health effects, and in three such cases, reported adverse health effects were associated with sites from which samples testing positive for STX had been received. Symptoms of fever, eye irritation, abdominal pains, and skin rash were reported in children aged 2 10 years after exposure to the water. These were not the adverse human symptoms typical of STX poisoning; rather, they represented acute effects often reported following recreational exposure to cyanobacterial blooms. PMID- 15892063 TI - Characterization of proteases in guts of Daphnia magna and their inhibition by Microcystis aeruginosa PCC 7806. AB - Many cyanobacteria produce peptides that inhibit mammalian proteases. The hypothesis that inhibitors of mammalian proteases produced by cyanobacteria also interfere with digestive proteases of natural cladoceran grazers was tested by comparing the effects of cyanobacterial protease inhibitors on digestive proteases from Daphnia magna and on commercially available bovine proteases. The major digestive proteases of D. magna are trypsins and chymotrypsins, which differ from those of bovine origin in substrate specificity and susceptibility to synthetic inhibitors. An extract from Microcystis aeruginosa strain PCC 7806 inhibited both types of D. magna proteases. Subsequent fractionation of the extract by high-performance liquid chromatography indicated that several inhibitors are produced by M. aeruginosa that differ in their specificity for the trypsins and chymotrypsins of D. magna. Two fractions differed in their inhibitory effect on proteases of D. magna and bovine origin; therefore, assessment of the impact of cyanobacterial protease inhibitors on natural communities requires the use of digestive proteases from ecologically relevant grazers. PMID- 15892064 TI - Influence of TIPS on development of hepatocellular carcinoma in cirrhosis. PMID- 15892065 TI - Activity and substrate specificity of cytosolic and microsomal glutathione S transferase in Australian black tiger prawns (Penaeus monodon) after exposure to cyanobacterial toxins. AB - Cyanobacterial toxins have been shown to have a far-reaching impact-from aquatic organisms to human health. Aquatic organisms are typically exposed in their natural environment to toxic cyanobacteria, and exposure can occur via ingestion of cyanobacterial cells or by bioaccumulation of water-borne toxin. The aquaculture and fisheries of crustaceans are among the most important seafood industries. Concomitant with the growth of this industry, the importance of the health of crustaceans increased. The black tiger prawn is the major cultivated prawn in Australia. The aquaculture of these prawns takes place in shallow ponds, where blooms, often of cyanobacteria, develop. Cyanobacterial toxins were hypothesized to contribute to the mortality of prawns. Many aquatic organisms have the possibility of detoxifying cyanobacterial toxins via conjugation to glutathione. The presence of several classes of the cytosolic glutathione S transferase system in black tiger prawns-mu, pi, theta, alpha, and tau-was shown using different substrates for measurement. Injection experiments with microcystin-LR and feeding experiments with nodularin revealed elevation of GST activity in different types of prawn tissue in parallel with reduction in the GST classes. Correlation analyses of toxin content of the prawns with GST activity showed that low toxin content was correlated with high elevation of enzymes and high toxin content with low elevation of enzymes. PMID- 15892066 TI - Benthic cyanobacteria from the Baltic Sea contain cytotoxic Anabaena, Nodularia, and Nostoc strains and an apoptosis-inducing Phormidium strain. AB - Benthic cyanobacteria from aquatic environments have been reported to produce biologically active metabolites. However, the toxicity and other biological activities of benthic cyanobacteria from the Baltic Sea are not well known. We determined the biological activities of 21 Anabaena, Calothrix, Nodularia, Nostoc, and Phormidium strains isolated from benthic littoral habitats of the Baltic Sea. We studied whether benthic cyanobacterial extracts caused cytotoxicity by necrosis or induced apoptosis in two mammalian cell lines, a human leukemia cell line (HL-60) and a mouse fibroblast cell line (3T3 Swiss), and examined potential hepatotoxin (microcystin and nodularin) production. Five of the six benthic Anabaena strains, one of the two Nostoc strains, and two of the three Nodularia strains were highly cytotoxic to human leukemia cells. The Calothrix and Phormidium strains did not cause LDH leakage, but the extract of Phormidium strain BECID15 induced apoptosis in the HL-60 cells. Neither the microcystin synthetase E (mcyE) nor the nodularin synthetase F (ndaF) gene was amplified by PCR, and no microcystins or nodularins were detected by the protein phosphatase inhibition assay from the cyanobacterial strains included in this study. This indicates that benthic Baltic cyanobacteria contain potentially harmful cytotoxic compounds even though they do not produce microcystins or nodularins. These cytotoxic compounds remain to be characterized, and the mechanisms of cytotoxicity need to be studied further. PMID- 15892067 TI - Organ distribution and bioaccumulation of microcystins in freshwater fish at different trophic levels from the eutrophic Lake Chaohu, China. AB - This article reports the organ distribution and bioaccumulation of hepatotoxic microcystins (MCs) in freshwater fishes at different trophic levels from the large, shallow, eutrophic Lake Chaohu in September 2003, when there were heavy surface blooms of toxic cyanobacteria. Among all fish, intestines and blood had the highest average content of MC-RR + MC-LR (22.0 and 14.5 microg g(-1) DW, respectively), followed by liver, bile, and kidney (7.77, 6.32, and 5.81 microg g(-1) DW, respectively), whereas muscle had the least (1.81 microg g(-1) DW). MC content in muscle was highest in carnivorous fish (Culter ilishaeformis, 2.22 microg g(-1) DW) and omnivorous fish (Carassius auratus, 1.96 microg g(-1) DW) and was lowest in phytoplanktivorous fish (Hypophthalmichthys molitrix, 1.65 microg g(-1) DW) and herbivorous fish (Parabramis pekinensis 0.660 microg g(-1) DW). However, the amount of MC in the gut of H. molitrix (137 microg g(-1) DW) was more than 20 times that in the other fish (<6.50 microg g(-1) DW). The MCs showed a tendency to accumulate up the food chain, and piscivorous fish at the top of the food chain were at high risk of exposure to MCs in Lake Chaohu. Our study is the first to report MC concentrations in the bile and blood of wild fish. One hundred grams of fish muscle would contain 2.64-49.7 microg of MC-LR equivalent, or about 1.3-25 times the recommended tolerable daily intake of MC-LR by humans, indicating that fish are already severely contaminated by MCs and that the local authorities should warn the public of the risk of poisoning by eating the contaminated fish. PMID- 15892068 TI - Spatial distribution and temporal variation of Microcystis species composition and microcystin concentration in Lake Biwa. AB - Spatial and temporal variation in Microcystis species composition and microcystin concentration, quantified by enzyme-linked immunosorbent assay and high performance liquid chromatography, were investigated during a 3-year period (1998 2000) in the Northern Basin of Lake Biwa. The Northern Basin generally had a concentration of 5 microg L(-1) or less, except at station 1 (Nagahama Bay) from July to October during the study period. The maximum concentration at station 1 was 22.7, 35.9, and 22.0 microg L(-1) in October of 1998, 1999, and 2000, respectively. Eleven species of cyanobacteria were observed: Microcystis aeruginosa, M. ichthyoblabe, M. novacekii, M. wesenbergii, Oscillatoria raciborskii, Anabaena oumiana, A. affinis, A. flos-aquae, A. ucrainica, A. smithii, and A. crassa. Of these, M. aeruginosa and M. wesenbergii were the main components observed. A high concentration of microcystin in the lake water was mostly a result of variation in the relative amount of toxic M. aeruginosa rather than of the total Microcystis cell number. This was supported by the analytical results for isolated strains. Microcystis spp. cell density in the Northern Basin appeared to increase gradually over the course of the study. This is the first study to have surveyed the Northern Basin of Lake Biwa, which supplies drinking water to 14 million people and is the largest lake in Japan. PMID- 15892069 TI - Occurrence of toxic blue-green algae in the Kucukcekmece lagoon (Istanbul, Turkey). AB - The concentration of microcystin (MC) in the Kucukcekmece Lagoon, Istanbul, Turkey, and the physicochemical and biological parameters of water quality were investigated from October 2000 to June 2003. Water samples were collected from surface waters at three sites. Most bloom samples were dominated by Microcystis aeruginosa. The major microcystin variants detected by HPLC-PDA were microcystin YR and microcystin-LR. Microcystin concentration increased dramatically from early summer to early autumn and thereafter tended to decrease. The toxin concentration found in the filtered samples from surface waters varied between 0.06 and 24.2 microg L(-1) microcystin-LR equivalents. Each year extensive fish mortality was recorded between mid-June and early October, coinciding with heavy algal blooms. A comparison of the conditions associated with cyanotoxin episodes in 2000, 2001, and 2002 showed that the microcystin increase was related to temperature, high concentration of dissolved nutrients, high light intensity (PAR). The highest MC concentrations were recorded at temperatures between 24 degrees C and 28.5 degrees C. Field data showed that the highest MC concentration (>3 microg L(-1)) and the highest cyanobacterial biomass (>30 mg L(-1)) corresponded to a total nitrogen:total phosphorus ratio greater than 7:1. The highest concentrations of M. aeruginosa biomass (173 mg L(-1)) and MC (24.2 microg L(-1) MC-LR equiv.) and the highest salinity (8.8%) were measured concurrently in the lagoon. To our knowledge, this is the first evidence of cyanobacterial toxins in the Kucukcekmece Lagoon. PMID- 15892070 TI - Microcystin content of Microcystis aeruginosa is modulated by nitrogen uptake rate relative to specific growth rate or carbon fixation rate. AB - Modulation of microcystin production has been extensively studied in both batch and continuous cultures. Positive correlations with medium nitrogen, medium phosphorous, light intensity, inorganic carbon availability, and growth rate have been reported. Negative correlations have been reported between microcystin content and medium phosphorous. The only reported quantitative relationship between any variable and microcystin production was that of growth rate. Microcystis aeruginosa PCC7806 was therefore cultured under continuous culture conditions in a bubble-lift reactor at a growth rate of 0.01 h(-1) in modified BG11 (constant phosphate concentration of 0.195 mM and varying nitrate from 0.125 to 18 mM) and sampled at steady states for analysis of cell number, microcystin content, cellular N and P, residual medium nutrient concentration, and carbon fixation rate. Cellular microcystin quotas showed significant positive correlation with both nitrate uptake and cellular nitrogen content and were negatively correlated with carbon fixation rate, phosphate uptake, and cellular phosphorous. Thus, the ratio of nitrate uptake to phosphate uptake, cellular N to cellular P, and nitrate uptake to carbon fixation were positively correlated to cellular microcystin. Microcystin quotas increased 10-fold from the lowest to the highest steady-state values. Cellular microcystin content therefore is controlled to a significant extent by variables other than growth rate, as was previously reported, with nitrogen the most significant modulator. Batch culture in BG11 under identical conditions yielded increased microcystin when nitrogen uptake exceeded relative growth rate, confirming the importance of nitrogen uptake in the modulation of microcystin content for a specific growth rate. PMID- 15892071 TI - Levels of microcystins in two Argentinean reservoirs used for water supply and recreation: differences in the implementation of safe levels. AB - Toxic cyanobacterial blooms are an increasing problem in Argentina. The production of cyanobacterial hepatotoxins (microcystins) and their presence in drinking and recreational waters represent a growing danger to human and animal health. Risk management deals with the probability that a certain exposure to toxins will lead to specific health outcomes. Various model schemes for risk management have been portrayed, most of which have some common elements. These include the need for an information base on which to make decisions. Thus, seasonal variability in the concentrations of total microcystins and cyanobacterial cells was studied in two reservoirs: San Roque and Paso de las Piedras. Both reservoirs are eutrophic water bodies and mainly used to supply drinking water and for recreation. Because San Roque is an important recreational spot, the spatial distribution of microcystins was also investigated. Sampling of the San Roque Reservoir occurred from 1998 to 2000 and of the Paso de las Piedras Reservoir from June to December 2002 (late autumn, winter, and spring). Microcystins were identified by LC-MS, and their concentrations were measured using the enzyme-linked immunosorbent assay. These hepatotoxic compounds were detected in all seasons, and even during winter relatively high concentrations were observed. Concentrations in the San Roque Reservoir varied from undetectable to 920 microg/L. On the contrary, in the Paso de las Piedras Reservoir the concentration remained below 1 microg/L, which is the provisional guideline value proposed by the World Health Organization (WHO; Chorus and Bartram, 1999) for microcystin-LR in drinking water. Comparison of cell number and concentration of total microcystins indicated that the phytoplankton in San Roque Reservoir contained more toxic cyanobacterial strains than did the Paso de las Piedras Reservoir. This indicates that the threshold of 2000 cell/mL proposed by WHO as a alert level should be adjusted: it should be reduced for the San Roque Reservoir, whereas it seems appropriate for the Paso de la Piedras Reservoir. PMID- 15892072 TI - Seasonal production and molecular characterization of microcystins in Oneida Lake, New York, USA. AB - Oneida Lake, northeast of Syracuse, New York, in the United States, is a shallow eutrophic lake with a well-established toxic cyanobacterial population. Samples for DNA, toxin, and phycological analyses were collected from six stations throughout the summers of 2002 (78 samples) and 2003 (95 samples). DNA was amplified by PCR using primer sets specific to the nonribosomal microcystin synthetase complex (mcyB and mcyD). PCR analysis in 2002 indicated that the microcystin genes were present in the water column from mid-June through October, as 88% of the samples tested positive for mcyB and 79% of the samples tested positive for mcyD. In both years the onset of microcystin production was detected as early as mid-July by the protein phosphatase inhibition assay, reaching a maximum in 2002 of 2.9 microg L(-1) and in 2003 of 3.4 microg L(-1). Beginning in mid- to late August of both years the microcystin level at all six stations was in excess of the World Health Organization (WHO) advisory level of 1.0 microg L( 1). In the present study we compared microcystin occurrence and potential production at the six stations using protein phosphatase inhibition assay, high performance liquid chromatography, and polymerase chain reaction analyses. PMID- 15892073 TI - Diversity of hepatotoxic microcystins and bioactive anabaenopeptins in cyanobacterial blooms from Greek freshwaters. AB - Microcystins (MCs) and anabaenopeptins from 26 cyanobacterial bloom samples dominated mainly by the genus Microcystis and collected from seven Greek freshwaters were identified and quantified by high-performance liquid chromatography coupled to a diode array detector. All the samples analyzed contained microcystins; in 27% of the samples anabaenopeptins were detected but not anabaenopeptilide (A). In each sample 1-7 microcystins and up to two anabaenopeptins (anabaenopeptins A and B) were identified. MC-RR and MC-LR were the predominant microcystins, followed by MC-YR. MC-LA and demethylated variants of MC-LR and MC-RR also were present but were not abundant. Total content of microcystin and anabaenopeptin varied from 40 to 2565 microg g(-1) freeze-dried material (mean 674.5 microg g(-1)) and from undetectable to 48 microg g(-1) freeze-dried material (mean 6.2 microg g(-1)), respectively. Qualitative and quantitative variation in the microcystins in the samples indicates there may be geographical trends in the distribution of microcystins. This study reports for the first time (1) the widespread occurrence of several different microcystins in Greek freshwaters and (2) quantitative data on the anabaenopeptins produced in natural cyanobacterial populations. PMID- 15892074 TI - Identification of potentially toxic environmental Microcystis by individual and multiple PCR amplification of specific microcystin synthetase gene regions. AB - Reliable cyanotoxin monitoring in water reservoirs is difficult because of, among other reasons, unpredictable changes in cyanobacteria biomass, toxin production, and inadequate sampling frequency. Therefore, it would be useful to identify potentially microcystin-producing strains of cyanobacterial populations in field samples. With this aim, we developed a methodology to distinguish microcystin producing from non-producing Microcystis strains by amplifying six characteristic segments of the microcystin synthetase mcy cluster, three corresponding to the nonribosomal peptide synthetase, genes mcyA, mcyB, and mcyC, and three to the polyketide synthase, genes mcyD, mcyE, and mcyG. For this purpose five new primer sets were designed and tested using purified DNA, cultured cells, and field colonies as DNA sources. Simultaneous amplification of several genes in multipex PCR reactions was performed in this study. The results obtained showed that: (i) the expected specific amplicons were obtained with all microcystin-producing strains but not with nonproducing strains; (ii) cells could be directly used as DNA templates, 2000 cells being a sufficient number in most cases; (iii) simultaneous amplification of several gene regions is feasible both with cultured cells and with field colonies. Our data support the idea that the presence of various mcy genes in Microcystis could be used as a criterion for ascribing potential toxigenicity to field strains, and the possibility of applying whole cell assays for the simultaneous amplification of various genes may contribute significantly to simplifying toxigenicity testing. PMID- 15892075 TI - Safety of short-term administration of celecoxib in decompensated cirrhosis. PMID- 15892076 TI - Lymph node metastasis in an animal model: effect of piecemeal laser surgical resection. AB - BACKGROUND AND OBJECTIVES: Endoscopic laser surgical resection of advanced squamous cell carcinoma (SCC) often requires division of the tumor into several pieces. It is unknown if this approach influences the incidence of regional and distant metastases. STUDY DESIGN/MATERIALS AND METHODS: In 143 rabbits VX2 SCC was induced. Eight days later the tumor was resected by two different methods. In the first group en bloc cold steel resection was performed. In the second group piecemeal laser resection was performed. On the 51th day the animals were sacrificed and examined for lymph node and distant metastases. RESULTS: After piecemeal laser resection 47.7% of the animals had lymph node metastases compared to 24.6% after en bloc resection (P = 0.01). The incidence of distant metastases did not differ for the two groups. CONCLUSIONS: In our model narrow margin piecemeal laser resection was associated with a higher incidence of metastases compared to wide en bloc surgical resection. The exact mechanism responsible for this increase is unclear. PMID- 15892077 TI - Genetic diversity of the toxic cyanobacterium Microcystis in Lake Mikata. AB - The aim of the present study was to clarify the bloom dynamics and community composition of hepatotoxin microcystin-producing and non-microcystin-producing Microcystis genotypes in the environment. In Lake Mikata (Fukui, Japan) from April 2003 to January 2004, seasonal variation in the number of cells with microcystin (mcy) genotypes and the genetic diversity of the total population were investigated using quantitative competitive PCR and a 16S rDNA clone library, respectively. Using competitive PCR, cells with mcyA genotypes were quantified in August and October, and the ratio of the number of these mcyA genotypes to colony-forming Microcystis cells was 0.37 and 2.37, respectively. The 16S rDNA clones obtained could be divided into 12 ribotypes: a-l. Sixty-one Microcystis strains isolated from Lake Mikata during the sampling period were subjected to toxicity tests using HPLC and ELISA, PCR-based detection of the mcyA gene, and sequence analysis of the 16S rDNA. All isolates could be differentiated into 11 ribotypes (a, b, d, f, h, i, and m-q). Ribotypes b, f, i, m, n, and p had at least one strain that was a microcystin producer. In natural communities ribotypes b and f accounted for 85% of the 16S rDNA clones in August, and ribotypes b and i accounted for 24% of the clones in October. Thus, in some bloom stages the presence of microcystin genotypes identified using the 16S rDNA clone library correlated with that of mcy genotypes determined using competitive PCR. PMID- 15892078 TI - Clinical outcomes and mechanism of action for rheopheresis treatment of age related macular degeneration (AMD). AB - The primary goals are to provide a comprehensive explanation of the potential role of therapeutic apheresis in the treatment of Age-Related Macular Degeneration (AMD). Initial clinical results with this technique and a summary of current literature that addresses the mechanism of action for the Rheopheresis approach are presented. Rheopheresis has been found to be a safe and effective application of double filtration plasmapheresis (DFPP) for extracorporeal hemorheotherapy. In this report, it is proposed that Rheopheresis results in an immediate decrease in the proportion of high molecular weight proteins that could combine with the TIMP-3 fibulin complex allowing for the barely functioning retinal pigment epithelial (RPE) cells to function better and diminish the release of vascular endothelial growth factor (VEGF). Interim results from the randomized, double-masked MIRA-1 clinical trial include (1) improved vision restoration; 28.0% of Treated Primary Eyes increased by > or = 2 lines of best corrected visual acuity (BCVA) compared to 18.2% of Placebo Eyes; (2) a decline in progressive vision loss; 0.0% of treated eyes progressing to worse than 20/200 vision over the 12-month study compared to 18.2% of Placebo Eyes; (3) 57.9% of Treatment Eyes obtained improvement in their BCVA to 20/40 or better (driver's license qualification), compared to only 14.3% of Placebo Eyes 12-month post treatment. Rheopheresis treatment shows strong promise as a viable clinical option for patients suffering from the dry form of AMD in terms of minimizing vision loss, vision restoration, and overall quality of life factors. Expanded clinical outcomes from the ongoing MIRA-1 clinical study will be valuable in the assessment of this new clinical tool for ophthalmic applications. PMID- 15892079 TI - Novel Caenorhabditis elegans unc-119 axon outgrowth defects correlate with behavioral phenotypes that are partially rescued by nonneural unc-119. AB - UNC-119 function is necessary for the correct development of the Caenorhabditis elegans nervous system. Worms mutant for unc-119 exhibit nervous system structural defects, including supernumerary axon branches, defasciculated nerve fibers, and choice point errors. Axons of both mechanosensory (ALM) and chemo- sensory (ASI) neurons have elongation defects within the nerve ring. Expressing unc-119 cDNA in mechanosensory neurons rescues the elongation defect of ALM axons, but expression in ASI neurons does not rescue ASI axon elongation defects. Neither gross movement nor dauer larva formation defects are rescued in either case. However, expressing a construct including introns under the control of the same promoters results in substantial rescue of phenotypic defects. In these cases reporter expression expands to tissues outside those specified by the promoter, notably into head muscles. Surprisingly, expressing an unc-119 cDNA construct under the control of a muscle-specific promoter fully rescues the dauer formation defect and substantially rescues movement. Thus, although UNC-119 normally acts in a cell-autonomous fashion, the cell-nonautonomous rescue of neural function suggests that it either acts at the cell surface or that it can be transported into the cell from the extracellular environment and play its normal role. PMID- 15892080 TI - Quality systems in automated plateletpheresis in hospital-based blood transfusion service in north India. AB - The issues of providing quality blood products and maintaining donor safety are primary aims of blood transfusion services. A comprehensive quality system should be in place to fulfill these aims, which can be attained through strict adherence to the established standard operating procedures (SOPs). The Drugs and Cosmetics Act of India, which controls the licensing of blood transfusion services, does not provide clear guidelines regarding plateletpheresis procedure. We, therefore, established our own SOP and operational flow chart for plateletpheresis that can be easily followed by other centers in India. A total of 100 plateletpheresis procedures performed using two cell separators (CS3000 Baxter Healthcare, Round Lake, IL; MCS3p, Haemonetics Corporation, Braintree, MA) were evaluated following our established SOP. The mean platelet yield in CS3000 was 2.9 +/- 0.84 x 10(11) and in MCS3p it was 2.88 +/- 0.75 x 10(11)per unit. However, only 4-7% of SDPs showed WBC levels <5 x 10(6) due to lack of appropriate methods to quantitate residual WBC counts. Six of 100 donors complained of hypocalcemic symptoms. The operational flow chart designed in this study was found to be simple and easy to adapt by blood transfusion services in this country. PMID- 15892081 TI - The natural history and role of immunoadsorption in dilated cardiomyopathy. PMID- 15892082 TI - Closed system generation of dendritic cells from a single blood volume for clinical application in immunotherapy. AB - Dendritic cells (DC) used for clinical trials should be processed on a large scale conforming to current good manufacturing practice (cGMP) guidelines. The aim of this study was to develop a protocol for clinical grade generation of immature DC in a closed-system. Aphereses were performed with the Cobe Spectra continuous flow cell separator and material was derived from one volume of blood processed. Optimisation of a 3-phase collection autoPBSC technique significantly improved the quality of the initial mononuclear cell (MNC) product. Monocytes were then enriched from MNC by immunomagnetic depletion of CD19+ B cells and CD2+ T cells and partial depletion of NK cells using the Isolex 300I Magnetic cell selector. The quality of the initial mononuclear cell product was found to determine the outcome of monocyte enrichment. Enriched monocytes were cultured in Opticyte gas-permeable containers using CellGro serum-free medium supplemented with GM-CSF and IL-4 to generate immature DC. A seeding concentration of 1 x 10(6) was found optimal in terms of DC phenotype expression, monocyte percentage in culture, and cell viability. The differentiation pattern favours day 7 for harvest of immature DC. DC recovery, viability, as well as phenotype expression after cryopreservation of immature DC was considered in this study. DC were induced to maturation and evaluated in FACS analysis for phenotype expression and proliferation assays. Mature DC were able to generate an allogeneic T-cell response as well as an anti-CMV response as detected by proliferation assays. These data indicate that the described large-scale GMP-compatible system results in the generation of stable DC derived from one volume of blood processed, which are qualitatively and quantitatively sufficient for clinical application in immunotherapeutic protocols. PMID- 15892083 TI - A case of non-lethal pulmonary air embolism after leukapheresis catheter removal. AB - Mononuclear cell leukapheresis requires good-quality venous access. Catheter placement and removal of the catheter may be associated with life-threatening local or systemic complications. Thus, prompt recognition of these complications and appropriate therapy can be life-saving. We report the case of a young man who presented with an air embolism following removal of a jugular venous catheter after peripheral blood stem cell collection. We have reviewed the signs and symptoms presented by the patient and the methodology used to remove the catheter. Catheter removal requires careful attention in order to avoid potentially serious complications. PMID- 15892084 TI - Treatment of acute occlusion of the retinal artery by LDL-apheresis. PMID- 15892085 TI - In vitro and in vivo evaluation of Adacolumn cytapheresis in healthy subjects. AB - Adacolumn is a medical device for adsorptive cytapheresis. It has been developed for selective adsorption of granulocytes and monocytes from peripheral blood of patients with immune disorders, such as autoimmune diseases and chronic inflammatory diseases. A double blind sham-controlled crossover study design was used in order to evaluate in vivo biological responses of leukocytes as well as biocompatibility during and after Adacolumn cytapheresis in healthy volunteers. In addition, experiments were undertaken to further evaluate leukocyte reactions to Adacolumn carrier (G-1: cellulose diacetate) beads in vitro. Six healthy volunteers, 4 males and 2 females, with a mean age of 26.7 years were randomly assigned to one of the two treatment arms in a crossover fashion. Three subjects received a single Adacolumn treatment, followed by a single sham treatment at an interval of 7 days. The other three subjects received the two treatments in reverse order. All subjects were followed up 7 days after the last treatment. Additionally, in vitro investigations were carried out using blood from the healthy donors to examine the effect of G-1 beads on granulocyte functions. In vitro exposure of human peripheral blood to G-1 beads caused downregulation of L selectin expression and upregulation of Mac-1 expression on granulocytes, leading to a marked reduction of adhesive capacity of granulocytes to endothelial cells. The exposure also led to decreased granulocyte chemotactic activity to IL-8. The number of granulocytes and monocytes clearly decreased during Adacolumn cytapheresis. Granulocytes showed marked phenotypic changes of L-selectin(Low) and Mac-1(Hi) after passing through Adacolumn in vivo. Expression of TNF-alpha and chemokine receptors was downregulated. In addition, TNF-alpha and IL-1beta producing capacity of peripheral blood leukocytes was decreased after Adacolumn cytapheresis and these changes lasted even one week after the cytapheresis. The level of complement fragments, C3a and C5a, increased, while bradykinin concentration did not change during Adacolumn cytapheresis. Exposure of human peripheral blood to G-1 beads, both in vitro and in vivo, caused a significant reduction of adhesive capacity and proinflammatory cytokine producing capacity of peripheral blood leukocytes. Such changes were not observed after sham apheresis. Despite complement activation, tolerability of Adacolum cytapheresis was not influenced. These findings may at least partly explain the beneficial effect of Adacolumn cytapheresis in the treatment of autoimmune diseases. PMID- 15892086 TI - Harvesting peripheral blood progenitor cells from healthy donors: retrospective comparison of filgrastim and lenograstim. AB - Mobilization of CD34+ into peripheral blood is attained by either glycosylated (lenograstim) or non-glycosylated recombinant G-CSF (filgrastim). 101 donors, 57 males, median age 42 years (range 16-63) entered this retrospective study. Group I (55 cases) received filgrastim and group II lenograstim subcutaneously for 5-6 days. The peak number of CD34+ cells/microl blood observed on day 4 and 5 was not significantly different in the two groups. No differences were shown in terms of both circulating CFU-GM at the time of harvesting and CD34+ target of collection. The most frequent side effects were bone pain (18.2% grade I; 36.4% grade II, 7.3% grade III), headache (18.2%), nausea (9.1%), fever (5.5%) and a mild splenomegaly (> 2 cm) (5.5%) in filgrastim group, and bone pain (37.0% grade I, 26.1% grade II, 2.2% grade III), headache (17.4%), nausea (15.2%), fever (4.4%) and a mild splenomegaly (4.3%) in lenograstim group, respectively. CD34+ collection was associated with thrombocytopenia, which was not significantly different between the two groups. No donor in either group developed long-term adverse effects. We conclude that both G-CSFs are comparable in terms of CD34+ cell collection, safety and tolerability. PMID- 15892088 TI - Optimizing peripheral stem cell mobilization and harvest in very small children. PMID- 15892087 TI - Reduced oxidative stress in parallel to improved cardiac performance one year after selective removal of anti-beta 1-adrenoreceptor autoantibodies in patients with idiopathic dilated cardiomyopathy: data of a preliminary study. AB - Patients with idiopathic dilated cardiomyopathy (IDC) were treated with selective immunoadsorption to remove anti-beta 1-adrenoreceptor autoantibodies (anti-beta1A AB). After one year, the effect on cardiac performance and oxidative stress was tested. Extracorporeal immunoadsorption of the whole IgG class in IDC patients for the removal of anti-beta1A-AB reduced oxidative stress in parallel to an improvement of cardiac performance. However, the non-specificity of IgG adsorption means that these beneficial effects cannot be attributed exclusively to anti-beta1A-AB removal. In an open clinical pilot study enrolling 8 patients with IDC prior to and one year after selective immunoadsorption of anti-beta1A AB, plasma markers for oxidative stress--thiobarbituric acid-reactive substances (TBARS), lipid peroxides (LPO) and anti-oxidized low-density lipoprotein autoantibodies (anti-oxLDL-AB)--were measured in parallel to evaluation of the left ventricular function using conventional echocardiography and wall motion analysis by tissue Doppler imaging. After one year, TBARS (Wilcoxon test with bootstrapping simulation for paired data: 95% confidence interval of the P value 0.020 to 0.029) and anti-oxLDL-AB (P = 0.025 to 0.035) were decreased in parallel to an improvement of the peak systolic wall motion velocity (P = 0.006 to 0.01) and left ventricular ejection fraction (P = 0.002 to 0.02). For changes over the study period, a direct correlation with borderline significance (P = 0.076) was calculated for TBARS to the left ventricular diameter in the diastole. One year after selective immunoadsorption for anti-beta1A-AB removal, patients with ICD show a reduction in oxidative stress and a parallel improvement in cardiac performance. PMID- 15892089 TI - Transfusion-transmitted Streptococcus pneumoniae from a single donor apheresis platelet unit. PMID- 15892090 TI - Daily cannabis smoking as a risk factor for progression of fibrosis in chronic hepatitis C. AB - Cannabinoids present in Cannabis sativa (marijuana) exert biological effects via cannabinoid receptors CB1 and CB2. We recently demonstrated that CB1 and CB2 receptors regulate progression of experimental liver fibrosis. We therefore investigated the impact of cannabis smoking on fibrosis progression rate in patients with chronic hepatitis C (CHC). Two hundred seventy consecutive untreated patients with CHC of known duration undergoing liver biopsy were studied. Demographic, epidemiological, metabolic, and virological data were recorded, and detailed histories of cannabis, alcohol, and tobacco use over the span of hepatitis C virus infection were obtained. Fibrosis stage, steatosis, and activity grades were scored according to Metavir system. Patients were categorized as noncannabis users (52.2%), occasional users (14.8%), or daily users (33.0%), and the relationship between cannabis use and fibrosis progression rate (FPR) or fibrosis stage was assessed. On multivariate analysis, six factors were independently related to a FPR greater than 0.074 (median value of the cohort): daily cannabis use (OR = 3.4 [1.5-7.4]), Metavir activity grade A2 or higher (OR = 5.4 [2.9-10.3]), age at contamination of more than 40 years (OR = 10.5 [3.0-37.1]), genotype 3 (OR = 3.4 [1.5-7.7]), excessive alcohol intake (OR = 2.2 [1.1-4.5]), and steatosis (OR = 2.0 [1.0-4.1]). Daily cannabis use was also an independent predictor of a rapid FPR (>0.15) (OR = 3.6 [1.5-7.5]). Finally, severe fibrosis (> or =F3) was also predicted by daily cannabis use (OR = 2.5 [1.1-5.6]; P = .034), independently of Metavir activity grade, excessive alcohol intake, age at liver biopsy, steatosis, and tobacco smoking. In conclusion, daily cannabis smoking is significantly associated with fibrosis progression during CHC. Patients with ongoing CHC should be advised to refrain from regular cannabis use. PMID- 15892091 TI - Autologous peripheral blood stem cell collections in children weighing less than 10 Kg with solid tumors: experience of a single center. AB - There have only been a few reports and limited performance of peripheral blood stem cell (PBSC) collection in very small children weighing less than 10 kg. In this study, we intended to evaluate the safety and yield of PBSC collection, with the efficacy of PBSC transplantation (PBSCT) in the smallest children with solid tumors. From January 1998 to February 2004, 173 children underwent PBSC collection in Samsung Medical Center, Korea. Of these, 15 (8.7%) children weighed less than 10 kg and their clinical diagnoses were neuroblastoma (10 cases), rhabdoid tumor (2 cases), rhabdomyosarcoma (2 cases), and Wilms tumor (1 case). PBSCs were collected following chemotherapy plus G-CSF mobilization. The median age and weight at the time of apheresis were 15 months and 9 kg, respectively. The median number of PBSC collection procedures per case was 4 (range, 2-7). The median cell yield per apheresis product was 0.95 (range, 0.01-33.32) x 10(6)/kg CD34+ cells and 1.96 (range, 0.12-23.39) x 10(8)/kg mononuclear cells. No complications associated with citrate toxicity and other adverse effect were observed during the procedures. After high-dose chemotherapy, 14 patients were reinfused with PBSCs alone and all showed successful hematopoietic recovery. We concluded that PBSC collection would be a safe and practical procedure, even when done in the smallest children, provided that adequate intravascular fluid volume and circulating red cell mass were maintained. Also, the use of PBSCs to support high-dose chemotherapy was well tolerated and might enhance hematological recovery in the smallest children showing the excellent efficacy of PBSCT. PMID- 15892092 TI - Robust ascertainment-adjusted parameter estimation. AB - Nonrandom ascertainment is commonly used in genetic studies of rare diseases, since this design is often more convenient than the random-sampling design. When there is an underlying latent heterogeneity, Epstein et al. ([2002] Am. J. Hum. Genet. 70:886-895) showed that it is possible to get unbiased or consistent estimation of population parameters under ascertainment adjustment, but Glidden and Liang ([2002] Genet. Epidemiol. 23:201-208) showed in a simulation study that the resulting estimates are highly sensitive to misspecification of the latent components. To overcome this difficulty, we consider a heavy-tailed model for latent variables that allows a robust estimation of the parameters. We describe a hierarchical-likelihood approach that avoids the integration used in the standard marginal likelihood approach. We revisit and extend the previous simulation, and show that the resulting estimator is efficient and robust against misspecification of the distribution of latent variables. PMID- 15892093 TI - Comparison of population- and family-based methods for genetic association analysis in the presence of interacting loci. AB - We compared different ascertainment schemes for genetic association analysis: affected sib-pairs (ASPs), case-parent trios, and unrelated cases and controls. We found, with empirical type 1 diabetes data at four known disease loci, that studies based on case-parent trios and on unmatched cases and controls often gave higher odds ratio estimates and stronger significance test values than ASP designs. We used simulations and a simplified disease model involving two interacting loci, one of large effect and one smaller, to examine interaction models that could cause such an effect. The different ascertainment schemes were compared for power to detect an effect when only the locus of smaller effect was genotyped. ASPs showed the greatest power for association testing under most models of interaction except under additive and certain epistatic crossover models, for which case/controls and case-parent trios did better. All ascertainment schemes gave an unbiased estimation of log genotype relative risks (GRRs) under a multiplicative model. Under nonmultiplicative interactions, GRRs at the minor locus as estimated from ASPs could be biased upwards or downwards, resulting in either an increase or decrease in power compared to the case/control or trio design. For the four known type 1 diabetes loci, we observed decreased risks with ASPs, which could be due to additive interactions with the remaining susceptibility loci. Thus, the optimal ascertainment strategy in genetic association studies depends on the unknown underlying multilocus genetic model, and on whether the goal of the study is to detect an effect or to accurately estimate the resulting disease risks. PMID- 15892094 TI - One shot laser pulse induced reversible spin transition in the spin-crossover complex [Fe(C4H4N2){Pt(CN)4}] at room temperature. PMID- 15892095 TI - Construction of substituted N-hydroxyindoles: synthesis of a nocathiacin I model system. PMID- 15892096 TI - Morphology and immunohistochemistry of efferent neurons of the goldfish corpus cerebelli. AB - In teleosts, cerebellar efferent neurons, known as eurydendroid cells, are dispersed within the cerebellar cortex rather than coalescing into deep cerebellar nuclei. To clarify their morphology, eurydendroid cells were labeled retrogradely by biotinylated dextran amine injection into the base of the corpus cerebelli. Labeling allowed the cells to be classified into three types-fusiform, polygonal, and monopolar-depending on their somal shapes and numbers of primary dendrites. The fusiform and polygonal type cells were distributed not only in the Purkinje cell layer but also in the molecular and granule cell layers. The monopolar type cells were distributed predominantly in the Purkinje cell layer of the ventrocaudal portion of the corpus cerebelli. These results suggest that there are some functional differences between these eurydendroid cell types. The eurydendroid cells were double-labeled by retrograde labeling and immunohistochemistry using specific antibodies against GABA, aspartate, and zebrin II. No GABA-like immunoreactivity was detected in the retrogradely labeled eurydendroid cells. About half of retrogradely labeled cells were immunoreactive to the anti-aspartate antibody, suggesting that some eurydendroid cells utilize aspartate as a neurotransmitter. Zebrin II reacts with cerebellar Purkinje cells but left all retrogradely labeled neurons nonreactive, although some of these were surrounded by immunopositive fibers. This relationship between the eurydendroid and Purkinje cells is similar to that between the deep cerebellar nuclei and Purkinje cells in mammals. PMID- 15892097 TI - mu-opioid receptor mRNA expression in identified hypothalamic neurons. AB - It has been known for a number of years that mu-opioid receptor agonists (e.g., morphine, beta-endorphin, and enkephalin) inhibit luteinizing hormone (LH), vasopressin (VP), and oxytocin (OT) release and stimulate prolactin secretion in rodents and primates by an action at the level of the brain. Also, electrophysiological studies have established that hypothalamic neurons, including gonadotropin-releasing hormone (GnRH), VP, OT, beta-endorphin, and dopamine neurons, are responsive to mu-receptor activation. Although mu-receptor expression has been demonstrated in the hypothalamus, there have been few studies localizing these receptors in neurosecretory neurons. Therefore, we sought to document mu-opioid receptor mRNA expression in immunocytochemically identified hypothalamic neurons. The brains from both female and male guinea pigs were examined by using in situ hybridization and immunocytochemistry. The studies revealed that mu-receptor mRNA was expressed in different diencephalic regions including the preoptic area, the bed nuclei stria terminalis, the paraventricular nucleus thalamus, and the anterior hypothalamus, as well as the supraoptic (SON), paraventricular (PVH), ventromedial, dorsomedial, and arcuate nuclei of the hypothalamus. Importantly, mu-opioid receptors were expressed in subpopulations of GnRH neurons (33.25 +/- 4.6% and 33.6 +/- 3.7% in females and males, respectively), dopamine neurons (51.7 +/- 5.8% to 75.0 +/- 2.6%, depending on neuronal location), beta-endorphin neurons (68.3.0 +/- 4.4%), and VP neurons (41 70%, depending on neuronal location). Because mu-opioid receptors couple via G proteins to activate inwardly rectifying potassium channels and to inhibit calcium channels, the presence of these receptors is likely to play a major role in directly controlling the excitability of hypothalamic neurons. PMID- 15892098 TI - Expression profiles of EphA3 at both the RNA and protein level in the developing mammalian forebrain. AB - The ephrin/Eph system is well known to regulate various aspects of brain development. In this study, we analyzed the expression profiles of EphA3 at both the RNA and protein level in developing mouse forebrains. Although the EphA3 gene is known to encode two isoforms of the receptors, a full-length transmembrane form, and a short, secretory form, only the full-length isoform was detected in the developing forebrain. We found that, in the early developmental stages, while EphA3 mRNA was expressed in the dorsal thalamus and the cortical intermediate zone (IMZ), the EphA3 protein was detected in the IMZ and the internal capsule, but not in the dorsal thalamus. In the later stages the mRNA was expressed in the most superficial region of the cortical plate, while the protein was expressed in the IMZ. This discrepancy between the mRNA and protein expression patterns might be attributed to the possibility of the protein being transported to the axons to regulate the thalamocortical and corticofugal projection. The results of double immunostaining for L1 and EphA3 or TAG-1 and EphA3 suggested that EphA3 protein was produced mainly in the thalamocortical axons and only partially in the corticofugal axons. In addition, the EphA3 protein was also detected in various other structures, such as the lateral olfactory tract, anterior commissure, and corpus callosum, suggesting the possibility that EphA3 might regulate the formation of various neuronal networks in the developing brain, including the TC projection and the commissural fibers. PMID- 15892099 TI - Anatomical abnormalities in dopaminoceptive regions of the cerebral cortex of dopamine D1 receptor mutant mice. AB - Alteration of dopamine neurotransmission during development can induce specific changes in neuronal structure and function. Here, we report specific morphological and neurochemical changes of projection neurons and interneurons of the medial frontal cortex of the dopamine D(1) receptor null mouse. Using immunostaining of cytoskeletal proteins and a crossbred D(1) receptor null:YFP transgenic reporter line, we demonstrate that the apical dendrites of pyramidal cells are abnormally organized in the prefrontal and anterior cingulate cortices of mice lacking the D(1) receptor. Neuronal processes exhibit a decrease in bundling and an increase in irregular, tortuous patterning as they weave a course towards the pial surface. In addition, there is increased parvalbumin staining of the dendrites of cortical interneurons in D(1) receptor null mice. Both pyramidal and interneuron alterations are evident by the early postnatal period and persist into adulthood. The alterations show regional specificity, in that dendritic profiles of projection neurons and interneurons in somatosensory and visual cortices develop normally. The abnormalities are reminiscent of those induced by prenatal exposure to cocaine in rabbits, an insult which has been shown to produce an attenuation of D(1) receptor-mediated responses through G(salpha). These results suggest that loss of D(1) receptor-mediated signaling during development produces permanent alterations in the cellular organization of specific cortical areas involved in attention, cognition, and emotion. Pharmacological and behavioral studies in the D(1) null mouse should be interpreted in the context of possible altered circuitry, given the presence of these developmental defects in the organization of dopaminoceptive regions of the cerebral cortex. PMID- 15892100 TI - Lamina-specific distribution of Synaptopodin, an actin-associated molecule essential for the spine apparatus, in identified principal cell dendrites of the mouse hippocampus. AB - Synaptopodin is an actin-associated molecule found in a subset of telencephalic spines. It is an essential component of the spine apparatus, a Ca(2+)-storing organelle and has been implicated in synaptic plasticity (Deller et al. [2003] Proc Natl Acad Sci U S A 100:10494-10499). In the rodent hippocampus, Synaptopodin is distributed in a characteristic region- and lamina-specific manner. To learn more about the cellular basis underlying this distribution, the regional, laminar, and cellular localization of Synaptopodin and its mRNA were analyzed in mouse hippocampus. First, Synaptopodin puncta densities were quantified after immunofluorescent labeling using confocal microscopy. Second, the dendritic distribution of Synaptopodin-positive puncta was studied using three-dimensional confocal reconstructions of Synaptopodin-immunostained and enhanced green fluorescence protein (EGFP)-labeled principal neurons. Synaptopodin puncta located within dendrites of principal neurons were primarily found in spines (>95%). Analysis of dendritic segments located in different layers revealed lamina-specific differences in the percentage of Synaptopodin positive spines. Densities ranged between 37% (outer molecular layer) and 14% (stratum oriens; CA1). Finally, synaptopodin mRNA expression was studied using in situ hybridization, laser microdissection, and quantitative reverse transcriptase polymerase chain reaction. Expression levels were comparable between all regions. These data demonstrate a lamina-specific distribution of Synaptopodin within dendritic segments of identified neurons. Within dendrites, the majority of Synaptopodin-positive puncta were located in spines where they represent spine apparatuses. We conclude, that this organelle is distributed in a region- and layer-specific manner in the mouse hippocampus and suggest that differences in the activity of afferent fiber systems could determine its distribution. PMID- 15892101 TI - Ltrk is differentially expressed in developing and adult neurons of the Lymnaea central nervous system. AB - The Trk receptor family plays diverse roles in both development and plasticity of the vertebrate nervous system. Ltrk is a related receptor that is expressed in the CNS of the mollusk Lymnaea, although little is known of its cellular distribution. This study provides three independent lines of evidence (based on RT-PCR, in situ hybridization, and immunohistochemistry) that Ltrk is universally expressed by neurons and dorsal body cells of both the juvenile and the adult Lymnaea CNS. The highest level of expression by neuronal somata occurs in the late juvenile stage, whereas axon collaterals express high levels throughout the animal's life span. Our data support multifunctional roles for Ltrk that parallel those of its mammalian counterparts. PMID- 15892102 TI - Topographic activation of the medial entorhinal cortex by presubicular commissural projections. AB - Previous investigations have shown that presubicular commissural fibers traveling in the caudal part of the dorsal hippocampal commissure (PSD) selectively activated the dorsalmost portion of the entorhinal cortex (EC), where they discharged perforant path neurons to the dorsal dentate gyrus. The dentate activation was followed by that of the dorsal hippocampus. The aim of the present study was to ascertain whether presubiculum commissural projections traveling in the PSD can also activate ventral levels of the EC and, if so, whether this activation is followed by that of the dentate gyrus-hippocampal system in the ventral hippocampus. The experiments were carried out in adult, anesthetized guinea pigs by field potential analysis. The results showed that presubicular fibers traveling at different PSD loci selectively activated specific EC portions, with caudal fibers activating only the dorsal EC and more rostral fibers activating ventral EC points. The region activated by PSD projections corresponded to the medial EC. Current source-density (CSD) analysis revealed that at both dorsal and ventral EC levels excitatory synaptic potentials followed by neuron discharge were generated in layer II, site of origin of the perforant path to the dentate gyrus. Activation of either dorsal or ventral levels of the EC was followed by activation of the dentate gyrus-hippocampal system in corresponding hippocampal segments. The results provide physiological evidence that the commissural presubicular projections activate the EC in a topographic manner. The massive activation of perforant path neurons at all EC levels suggests that presubicular signals may strongly influence the functions played by the EC-dentate-hippocampal system. PMID- 15892103 TI - Feedback connections to ferret striate cortex: direct evidence for visuotopic convergence of feedback inputs. AB - Interareal feedback connections are a fundamental aspect of cortical architecture, yet many aspects of their organization and functional relevance remain poorly understood. Previous studies have investigated the topography of feedback projections from extrastriate cortex to macaque area 17. We have extended this analysis to the ferret. We made restricted injections of cholera toxin B (CTb) into ferret area 17 and mapped the distribution of retrogradely labeled cells in extrastriate cortex. In addition to extensive label spreading within area 17, we found dense cell label in areas 18, 19, and 21 and the suprasylvian cortex and sparser connections from the lateral temporal and posterior parietal cortex. We made extensive physiological assessments of magnification factors in the extrastriate visual cortex and used these measures to convert the spread of labeled cortex in millimeters into a span in degrees of visual field. We also directly measured the visuotopic extents of receptive fields in the regions containing labeled cells in cases in which we made both CTb injections and physiological recordings in the same animals; we then compared the aggregate receptive field (ARF) of the labeled region in each extrastriate area with that of the injection site. In areas 18, 19, and 21, receptive fields of cells in regions containing labeled neurons overlapped those at the injection site but spanned a greater distance in visual space than the ARF of the injection site. The broad visuotopic extent of feedback connections is consistent with the suggestion that they contribute to response modulation by stimuli beyond the classical receptive field. PMID- 15892104 TI - High-resolution detection and mapping of genomic DNA alterations in neuroblastoma. AB - We used array-based comparative genomic hybridization (aCGH) to measure genomic copy number alterations (CNAs) in 42 neuroblastoma cell lines with known 1p36.3, 2p24 (MYCN), 11q23, and 17q23 allelic status. All cell lines showed CNAs, with an average of 22.0% of the genome of each sample showing evidence of gain (11.6%) or loss (10.4%). MYCN amplification was detected in 81% of cell lines, but other regions with high-level genomic amplification were observed only rarely. Gain of 17q material was present in 75% of the samples, and four discrete genomic regions at 17q23.2-17q25.3 were defined. Novel regions of gain were identified, including a 2.6-Mb subtelomeric region at 5p that includes the telomerase reverse transcriptase gene (TERT), which was found in 45% of the cell lines. Hemizygous deletions were noted at 1p36.23-1p36.32 and 11q23.3-11q25 in 60% and 36%, respectively, of the samples, with other frequent (>25%) regions of deletion localized to 1p32.1, 3p21.31-3p22.1, 5q35.2-5q35.3, 7q31.2, 7q34, 9q22.3-9q24.1, 10q26.11-10q26.12, 16q23.1-16q24.3, 18q21.32-18q23, and 20p11.21-20p11.23. A smallest region of overlap (SRO) for CNAs was mapped across all experiments and in each case was consistent with or refined the published data. A single cell line showed a homozygous deletion at 3p22.3, which was verified, and this location was refined by FISH and PCR. There was outstanding concordance of aCGH with PCR-based CNA detection methods. Several potential cooperating loci were identified, including deletion of 11q23-25, which was highly associated with both regional gain and loss at multiple chromosomal loci but was inversely correlated with the deletion of 1p36. Taking all of this together indicates that aCGH can accurately measure CNAs in the neuroblastoma genome and facilitate gene discovery efforts by high-throughput refinement of candidate loci. PMID- 15892105 TI - Mitochondrial DNA mutations and mitochondrial DNA depletion in gastric cancer. AB - Gastric carcinoma is one of the most common types of cancer in Taiwan. Somatic mitochondrial DNA (mtDNA) alteration in gastric carcinoma and its association with clinicopathologic features remain unclear. When we used polymerase chain reaction (PCR) and direct sequencing, 15 of the 31 (48%) gastric carcinomas displayed somatic mutations in the D-loop region, a hot spot for mutations in mtDNA of human cancers. Ten (67%) cancers with the somatic mutations in the D loop had insertion or deletion mutations in nucleotide position (np) 303-309 in the mononucleotide repeat region. One carcinoma carried tandem duplication and triplication flanked by mononucleotide repeats starting at np 311 and 568, respectively, in the D-loop. We also detected the common 4,977-bp deletion in 17 (55%) of the noncancerous tissue samples, but only in three (9%) carcinomas. Moreover, we quantified the mtDNA content using a competitive PCR technique and found that mtDNA depletion occurred in 17 (55%) of the gastric carcinomas. Although no significant association was found between clinicopathologic features and the mtDNA mutations in the D-loop, mtDNA depletion was observed significantly in the ulcerated, infiltrating (Borrmann's type III) and diffusely thick (Borrmann's type IV) types of gastric carcinomas (P = 0.018). Our results suggest that somatic mtDNA mutations and mtDNA depletion occur in gastric cancer and that mtDNA depletion is involved in carcinogenesis and/or cancer progression of gastric carcinoma. PMID- 15892106 TI - Cell polarity: don't forget calcium's role. PMID- 15892108 TI - Discontinuous or semi-discontinuous DNA replication in Escherichia coli? AB - The postulate that a stalled/collapsed replication fork will be generated when the replication complex encounters a UV-induced lesion in the template for leading-strand DNA synthesis is based on the model of semi-discontinuous DNA replication. A review of existing data indicates that the semi-discontinuous DNA replication model is supported by data from in vitro studies, while the discontinuous DNA replication model is supported by in vivo studies in Escherichia coli. Until the question of whether DNA replicates discontinuously in one or both strands is clearly resolved, any model building based on either one of the two DNA replication models should be treated with caution. PMID- 15892107 TI - Adacolumn for selective leukocytapheresis as a non-pharmacological treatment for patients with disorders of the immune system: an adjunct or an alternative to drug therapy? AB - Inflammatory and/or autoimmune diseases like ulcerative colitis (UC) or Crohn's disease (CD) are debilitating chronic disorders that poorly respond to pharmacological interventions. Further, drug therapy has adverse effects that add to disease complications. The current thinking is that disorders like inflammatory bowel disease (IBD) reflect an over exuberant immune activation driven by cytokines including TNF-alpha. Major sources of cytokines include myeloid leukocytes (granulocytes, monocytes/macrophages), which in IBD are elevated with activation behavior and are found in vast numbers within the inflamed intestinal mucosa. Accordingly, myeloid cells should be the targets of therapy. Adacolumn is filled with cellulose acetate beads that selectively adsorb and deplete myeloid cells and a small fraction of lymphocytes (FcgammaR and complement receptors bearing cells). In one study, 20 steroid naive patients with moderate (n = 14) or severe (n = 6) UC according to Rachmilewitz despite 1.5-2.25 g/day of 5-aminosalicylic acid received 6 to 10 Adacolumn sessions at 2 sessions/week. Efficacy was assessed 1 week after the last session. The majority of patients responded to 6 sessions, 17 (85%) achieved remission. In 2 of the 3 non-responders, CAI was 8 and 12 in 1; all 3 had deep colonic ulcers at study initiation. Decreases were seen in total leukocytes (P = 0.003), % neutrophils (P = 0.003), % monocytes (P = 0.004), an increase in lymphocytes (P = 0.001), decreases in C-reactive protein (P = 0.0002), and rises in blood levels of soluble TNF-alpha receptors I (P = 0.0007), II (P = 0.0045). In a separate study, a case with very severe steroid refractory UC who received up to 11 sessions responded well and avoided colectomy. Further, myeloid cell purging with Adacolumn has been associated with the release of IL-1 receptor antagonist, suppression of TNF-alpha, IL-1beta, IL-6, IL-8, down-modulation of L-selectin and the chemokine receptor CXCR3. In conclusion, selective depletion of myeloid cells appears to induce anti-inflammatory effects and represents a non-pharmacological treatment for patients with active IBD. The treatment has a clear drug-sparing role. Changes in blood levels of inflammatory and anti-inflammatory factors are thought to contribute to the efficacy of this procedure. PMID- 15892109 TI - Integrative biology of sticky feet in geckos. AB - Geckos have gained ecological access to novel microhabitats by exploiting intermolecular van der Waals forces, which allow them to climb smooth vertical surfaces. They use microscopic surface-based phenomena to thrive in a macroscopic mass- and kinetic energy-based world. Here we detail this as a premier example of integrative biology, spanning seven orders of magnitude and a lot of interesting biology. Emergent properties arising from molecular adhesion include several adaptive radiations that have produced a great diversity of geckos worldwide. PMID- 15892110 TI - Many roads to resistance: how invertebrates adapt to Bt toxins. AB - The Cry family of Bacillus thuringiensis insecticidal and nematicidal proteins constitutes a valuable source of environmentally benign compounds for the control of insect pests and disease agents. An understanding of Cry toxin resistance at a molecular level will be critical to the long-term utility of this technology; it may also shed light on basic mechanisms used by other bacterial toxins that target specific organisms or cell types. Selection and cross-resistance studies have confirmed that genetic adaptation can elicit varying patterns of Cry toxin resistance, which has been associated with deficient protoxin activation by host proteases, and defective Cry toxin-binding cell surface molecules, such as cadherins, aminopeptidases and glycolipids. Recent work also suggests Cry toxin resistance may be induced in invertebrates as an active immune response. The use of model invertebrates, such as Caenorhabditis elegans and Drosophila melanogaster, as well as advances in insect genomics, are likely to accelerate efforts to clone Cry toxin resistance genes and come to a detailed and broad understanding of Cry toxin resistance. PMID- 15892111 TI - Coronin proteins as multifunctional regulators of the cytoskeleton and membrane trafficking. AB - Coronins constitute an evolutionarily conserved family of WD-repeat actin-binding proteins, which can be clearly classified into two distinct groups based on their structural features. All coronins possess a conserved basic N-terminal motif and three to ten WD repeats clustered in one or two core domains. Dictyostelium and mammalian coronins are important regulators of the actin cytoskeleton, while the fly Dpod1 and the yeast coronin proteins crosslink both actin and microtubules. Apart from that, several coronins have been shown to be involved in vesicular transport. C. elegans POD-1 and Drosophila coro regulate the actin cytoskeleton, but also govern vesicular trafficking as indicated by mutant phenotypes. In both organisms, defects in cytoskeleton and trafficking lead to severe developmental defects ranging from abnormal cell division to aberrant formation of morphogen gradients. Finally, mammalian coronin 7 appears not to execute any cytoskeleton related functions, but rather participates in regulating Golgi trafficking. Here, we review recent data providing more insight into molecular mechanisms underlying the regulation of F-actin structures, cytoskeletal rearrangements and intracellular membrane transport by coronin proteins and the way that they might link cytoskeleton with trafficking in development and disease. PMID- 15892112 TI - Coding repeats and evolutionary "agility". AB - The rapid generation of new shapes observed in the living world is the result of genetic variation, especially in "morphological" developmental genes. Many of these genes contain coding tandem repeats. Fondon and Garner have shown that expansions and contractions of these repeats are associated with the great diversity of morphologies observed in the domestic dog, Canis familiaris. In particular, they found that the repeat variations in two genes were significantly associated with changes in limb and skull morphology. These results open the possibility that such a mechanism contributes to the diversity of life. PMID- 15892113 TI - Shugoshin: a centromeric guardian senses tension. AB - To ensure accurate chromosome segregation during mitosis, the spindle checkpoint monitors chromosome alignment on the mitotic spindle. Indjeian and colleagues have investigated the precise role of the shugoshin 1 protein (Sgo1p) in this process in budding yeast. The Sgo proteins were originally identified as highly conserved proteins that protect cohesion at centromeres during the first meiotic division. Together with other recent findings, the study highlighted here has identified Sgo1 as a component that informs the mitotic spindle checkpoint when spindle tension is perturbed. This discovery has provided a molecular link between sister chromatid cohesion and tension-sensing at the kinetochore microtubule interface. PMID- 15892114 TI - Speciation patterns and mechanisms: a symposium to honor Ernst Mayr. PMID- 15892115 TI - Behavioural genetics. PMID- 15892116 TI - Traversing the conceptual divide between biological and statistical epistasis: systems biology and a more modern synthesis. AB - Epistasis plays an important role in the genetic architecture of common human diseases and can be viewed from two perspectives, biological and statistical, each derived from and leading to different assumptions and research strategies. Biological epistasis is the result of physical interactions among biomolecules within gene regulatory networks and biochemical pathways in an individual such that the effect of a gene on a phenotype is dependent on one or more other genes. In contrast, statistical epistasis is defined as deviation from additivity in a mathematical model summarizing the relationship between multilocus genotypes and phenotypic variation in a population. The goal of this essay is to review definitions and examples of biological and statistical epistasis and to explore the relationship between the two. Specifically, we present and discuss the following two questions in the context of human health and disease. First, when does statistical evidence of epistasis in human populations imply underlying biomolecular interactions in the etiology of disease? Second, when do biomolecular interactions produce patterns of statistical epistasis in human populations? Answers to these two reciprocal questions will provide an important framework for using genetic information to improve our ability to diagnose, prevent and treat common human diseases. We propose that systems biology will provide the necessary information for addressing these questions and that model systems such as bacteria, yeast and digital organisms will be a useful place to start. PMID- 15892117 TI - The centipede Strigamia maritima: what it can tell us about the development and evolution of segmentation. AB - One of the most fundamental features of the body plan of arthropods is its segmental design. There is considerable variation in segment number among arthropod groups (about 20-fold); yet, paradoxically, the vast majority of arthropod species have a fixed number of segments, thus providing no variation in this character for natural selection to act upon. However, the 1000-species strong centipede order Geophilomorpha provides an exception to the general rule of intraspecific invariance in segment number. Members of this group, and especially our favourite animal Strigamia maritima, may thus help us to understand the evolution of segment number in arthropods. Evolution must act by modifying the formation of segments during embryogenesis. So, how this developmental process operates, in a variable-segment-number species, is of considerable interest. Strigamia maritima turns out to be a tractable system both at the ecological level of investigating differences in mean segment number between populations and at the molecular level of studying the expression patterns of developmental genes. Here we report the current state of play in our work on this fascinating animal, including our recent finding of a double-segment periodicity in the expression of two Strigamia segmentation genes, and its possible implications for our understanding of arthropod segmentation mechanisms in general. PMID- 15892118 TI - Is ectopic expression caused by deregulatory mutations or due to gene-regulation leaks with evolutionary potential? AB - It has long been thought that gene expression is tightly regulated in multicellular eukaryotes, so that expression profiles match functional profiles. This conception emerged from the assumption that gene activity is synonymous with gene function. This paradigm was first challenged by comparative protein electrophoresis studies showing extensive differences in expression patterns among related species. The paradigm is now being challenged by evolutionary transcriptomics using microarray technologies. Most gene expression profiles display features that lack any obvious functional significance. The so-called "ectopic" expression refers to the expression of genes at times and locations where the target gene is not known to have a function. However, ectopic expression might be associated with genuine function even if this function is not essential or has yet to be ascertained. Alternatively, ectopic expression might come about as a superfluous by-product of regulatory systems, which would call for a revision of prevailing ideas about the specificity of gene regulation. We herein review available evidence for ectopic expression and the hypotheses proposed for its origin and evolution. We propose that ectopic expression must be regarded as part of an integrated phenotypic whole. It seems likely that ectopic expression represents a leak in the evolution of regulatory systems, but one that is endowed with considerable evolutionary possibilities. PMID- 15892119 TI - Rho GTPase expression in tumourigenesis: evidence for a significant link. AB - Rho proteins belong to the small GTPases superfamily. They function as molecular switches that, in response to diverse stimuli, control key signaling and structural aspects of the cell. Although early studies proposed a role for Rho GTPases in cellular transformation, this effect was underestimated due to the fact that no genetic mutations affecting Rho-encoding genes were found in tumors. Recently, it has become evident that Rho GTPases participate in the carcinogenic process by either overexpression of some of the members of the family with oncogenic activity, downmodulation of other members with suggested tumor suppressor activity, or by alteration of upstream modulators or downstream effectors. Thus, alteration of the levels of expression of different members of the family of Rho GTPases has been detected in many types of human tumors leading to a great interest in the cellular effects elicited by these oncoproteins. This essay reviews the current evidence of dysregulation of Rho signaling by overexpression in human tumors. PMID- 15892121 TI - Phase equilibrium in poly(rA).poly(rU) complexes with Cd2+ and Mg2+ ions, studied by ultraviolet, infrared, and vibrational circular dichroism spectroscopy. AB - Ultraviolet (UV) and infrared (IR) absorption and vibrational circular dichroism (VCD) spectroscopy were used to study conformational transitions in the double stranded poly(rA). poly(rU) and its components-single-stranded poly(rA) and poly(rU) in buffer solution (pH 6.5) with 0.1M Na+ and different Mg2+ and Cd2+ (10(-6) to 10(-2) M) concentrations. Transitions were induced by elevated temperature that changed from 10 up to 96 degrees C. IR absorption and VCD spectra in the base-stretching region were obtained for duplex, triplex, and single-stranded forms of poly(rA) . poly(rU) at [Mg2+],[Cd2+]/[P] = 0.3. For single-stranded polynucleotides, the kind of conformational transition (ordering -> disordering --> compaction, aggregation) is conditioned by the dominating type of Me2+-polymer complex that in turn depends on the ion concentration range. The phase diagram obtained for poly(rA) . poly(rU) has a triple point ([Cd2+] approximately 10(-4)M) at which the helix-coil (2 --> 1) transition is replaced with a disproportion transition 2AU --> A2U + poly(rA) (2 --> 3) and the subsequent destruction of the triple helix (3 --> 1). The 2 --> 1 transitions occur in the narrow temperature interval of 2 degrees -5 degrees . Unlike 2 --> 1 and 3 --> 1 melting, the disproportion 2 --> 3 transition is a slightly cooperative one and observed over a wide temperature range. At [Me2+] approximately 10(-3) M, the temperature interval of A2U stability is not less than 20 degrees C. In the case of Cd2+, it increases with the rise of ion concentration due to the decrease of T(m) (2-->3). The T(m) (3-->1) value is practically unchanged up to [Cd2+] approximately 10(-3)M. Differences between diagrams for Mg(2+) and Cd2+ result from the various kinds of ion binding to poly(rA).poly-(rU) and poly(rA). PMID- 15892122 TI - Evidence for creatine biosynthesis in Muller glia. AB - In high-energy metabolic tissues like the retina, creatine may play an important role in energy storage and in transmission of phosphate-bound energy substrates. To prove this, we investigated creatine synthesis in Muller glia. We also characterized the localization of the creatine synthetic enzyme, S-adenosyl-L methionine:N-guanidinoacetate methyltransferase (GAMT) in the retina. Reverse transcription-polymerase chain reaction analysis revealed that L-arginine:glycine amidinotransferase and GAMT mRNAs were expressed in the retina and the Muller cell line, TR-MUL5. [(14)C]Creatine was detected after incubation of isolated rat retina or TR-MUL5 cells with [(14)C]glycine, L-arginine and L-methionine, suggesting creatine synthesis in Muller glia. Western blot analysis also revealed expression of GAMT protein in the rat retina and TR-MUL5 cells. Furthermore, confocal immunofluorescent microscopy of dual-labeled rat retinal sections demonstrated co-localization of GAMT with glutamine synthetase. Taken together, the results of the present study indicate creatine synthesis in Muller glia, implying an important role of creatine in energy metabolism in the retina. PMID- 15892123 TI - Tenascin C induces a quiescent phenotype in cultured adult human astrocytes. AB - Astrocytic scar formation occurs subsequent to brain and spinal cord injury and impedes repair. The exact mechanisms of scar formation have yet to be elucidated but it is known that astrocytes within the scar have a different antigenic phenotype from normal or reactive astrocytes. Astrocyte cell culture offers a suitable system to identify factors that induce the scar phenotype as well as factors that reverse this process and that may help identify therapeutic strategies to treat astrogliosis. However, when placed in standard culture conditions, astrocytes become activated/reactive and express molecules characteristic of scar tissue in vivo. In the present study, we made use of this phenomenon to identify culture conditions that change the activated phenotype of cultured astrocytes into one characteristic of normal quiescent astrocytes. In particular, we examined the effect of extracellular matrix (ECM) proteins found in the human brain, on the phenotype of human adult astrocytes. Significantly fewer astrocytes expressed scar properties when grown on tenascin-C (TN-C) than those cultured on other ECM proteins or poly-L-lysine-coated dishes. TN-C also significantly reduced the proliferation rate of the astrocytes in vitro. In addition, further manipulation of culture conditions induced partial astrocyte reactivation. Our findings suggest that astrocytes grown on TN-C revert to a quiescent, nonactivated state that is partially reversible. This raises the possibility that therapeutic strategies aimed at manipulating TN-C levels during CNS injury may help reduce astrocytic scarring. PMID- 15892124 TI - Shape-persistent phenylene-acetylene macrocycles: large rings-low yield? PMID- 15892125 TI - Nucleoside transporter expression and function in cultured mouse astrocytes. AB - Uptake of purine and pyrimidine nucleosides in astrocytes is important for several reasons: (1) uptake of nucleosides contributes to nucleic acid synthesis; (2) astrocytes synthesize AMP, ADP, and ATP from adenosine and GTP from guanosine; and (3) adenosine and guanosine function as neuromodulators, whose effects are partly terminated by cellular uptake. It has previously been shown that adenosine is rapidly accumulated by active uptake in astrocytes (Hertz and Matz, Neurochem Res 14:755-760, 1989), but the ratio between active uptake and metabolism-driven uptake of adenosine is unknown, as are uptake characteristics for guanosine. The present study therefore aims at providing detailed information of nucleoside transport and transporters in primary cultures of mouse astrocytes. Reverse transcription-polymerase chain reaction identified the two equilibrative nucleoside transporters, ENT1 and ENT2, together with the concentrative nucleoside transporter CNT2, whereas CNT3 was absent, and CNT1 expression could not be investigated. Uptake studies of tritiated thymidine, formycin B, guanosine, and adenosine (3-s uptakes at 1-4 degrees C to study diffusional uptake and 1-60-min uptakes at 37 degrees C to study concentrative uptake) demonstrated a fast diffusional uptake of all four nucleosides, a small, Na(+) independent and probably metabolism-driven uptake of thymidine (consistent with DNA synthesis), larger metabolism-driven uptakes of guanosine (consistent with synthesis of DNA, RNA, and GTP) and especially of adenosine (consistent with rapid nucleotide synthesis), and Na(+)-dependent uptakes of adenosine (consistent with its concentrative uptake) and guanosine, rendering neuromodulator uptake independent of nucleoside metabolism. Astrocytes are accordingly well suited for both intense nucleoside metabolism and metabolism-independent uptake to terminate neuromodulator effects of adenosine and guanosine. PMID- 15892126 TI - Excitotoxic oligodendrocyte death and axonal damage induced by glutamate transporter inhibition. AB - Glutamate uptake is crucial to terminate glutamate signaling and to prevent excitotoxicity. The present study describes the expression of functional glutamate transporters GLAST and GLT-1 in oligodendrocytes by means of electrophysiology, uptake assays, and immunocytochemistry. Inhibition of glutamate uptake, both in oligodendrocyte cultures and in isolated optic nerves, increases glutamate levels and causes oligodendrocyte excitotoxicity, which is prevented by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and kainate receptor antagonists. Furthermore, glutamate transporter inhibitors or antisense oligonucleotides applied onto the optic nerve in vivo lead to oligodendroglial loss, massive demyelination, and severe axonal damage. Overall, these results demonstrate that the integrity of oligodendrocytes and white matter depends on proper glutamate transporter function. Deregulated transporter activity may contribute to acute and chronic white matter damage. PMID- 15892127 TI - Cluster disorder and ordering principles in Al-stabilized "LaI". PMID- 15892128 TI - Microfluidics with droplets. PMID- 15892129 TI - Overexpression of human transferrin in two oligodendroglial cell lines enhances their differentiation. AB - We have previously demonstrated that the addition of apotransferrin (aTf) to oligodendroglial cell (OLGc) primary cultures accelerates their maturation. Cells treated with aTf developed a multipolar morphology and displayed increased expression of mature OLGc markers. In this work, we studied the effect of Tf overexpression in two OLGc lines, N19 and N20.1. The former cells exhibit characteristics of OLGc precursors (O2A), while N20.1 cells express markers of more mature OLGcs. Using the complete cDNA of the human Tf gene, we obtained clones overexpressing Tf in both cell lines. These clones were evaluated for the expression of OLGc differentiation markers. In agreement with our previous results, we found that in the cells overexpressing Tf, there was an increased O(4), GC, and MBP immunoreactivity. To study the myelinogenic potential of these cells, we co-cultured N19 and N20.1 Tf-transfected cells together with cortical neurons. There was a dramatic increase in the morphological differentiation of the OLGcs accompanied by enhanced GC and MBP expression. The OLGcs appeared to establish contact with neurites and extend their processes along them. Only two MBP isoforms were detected in Tf-overexpressing clones, while all the isoforms were present in the co-cultures, suggesting that there was a modulation of MBP expression by neurons. Concomitantly, we found an increase in several proteins involved in axon-glia interaction, such as MAG, N-CAM, and F3/Contactin. This co culture system represents a potentially powerful tool to study neuron-glia interactions that occur during myelinogenesis and the role of Tf in this process. PMID- 15892130 TI - Disruption of the hyaluronan-based extracellular matrix in spinal cord promotes astrocyte proliferation. AB - Astrocyte proliferation is tightly controlled during development and in the adult nervous system. In the present study, we find that a high-molecular-weight (MW) form of the glycosaminoglycan hyaluronan (HA) is found in rat spinal cord tissue and becomes degraded soon after traumatic spinal cord injury. Newly synthesized HA accumulates in injured spinal cord as gliosis proceeds, such that high-MW HA becomes overabundant in the extracellular matrix surrounding glial scars after 1 month. Injection of hyaluronidase, which degrades HA, into normal spinal cord tissue results in increased numbers of glial fibrillary acidic protein (GFAP) positive cells that also express the nuclear proliferation marker Ki-67, suggesting that HA degradation promotes astrocyte proliferation. In agreement with this observation, adding high- but not low-MW HA to proliferating astrocytes in vitro inhibits cell growth, while treating confluent, quiescent astrocyte cultures with hyaluronidase induces astrocyte proliferation. Collectively, these data indicate that high-MW HA maintains astrocytes in a state of quiescence, and that degradation of HA following CNS injury relieves growth inhibition, resulting in increased astrocyte proliferation. PMID- 15892131 TI - Asymmetric synthesis of the 1-epi aglycon of the cripowellins A and B. PMID- 15892132 TI - Direct asymmetric alpha-fluorination of aldehydes. PMID- 15892133 TI - Preface. PMID- 15892134 TI - Dendritic spine abnormalities in the occipital cortex of C57BL/6 Fmr1 knockout mice. AB - Fragile X syndrome (FXS) is the most common form of inherited mental retardation. Observed neuropathologies associated with FXS include abnormal length, morphology, and density of dendritic spines, reported in individuals with FXS and in Fmr1 knockout (KO) mice, an animal model of FXS. To date, however, these neuropathologies have been studied in Fmr1 KO mice bred in a FVB background (a strain with genetic mutations that complicate interpretation of results) and findings have been inconsistent. Here, Golgi-Cox impregnation was used to investigate length, morphology, and density of dendritic spines on layer V pyramidal neurons in visual cortices of Fmr1 KO and wildtype (WT) mice bred in a C57BL/6 background. We report that spine abnormalities in these animals parallel abnormalities reported in humans with FXS, perhaps to a greater degree than KO mice bred in an FVB background. Specifically, Fmr1 KO mice bred in a C57BL/6 background exhibited significantly more longer dendritic spines and fewer shorter spines, as well as more spines with immature-appearing morphology and fewer with mature-appearing morphology than WT littermates. Spine length abnormalities were demonstrated to be largely independent of spine morphology abnormalities, as the length phenotype was observed in KOs even within a morphological category. Fmr1 KO mice also had a greater overall spine density than WTs. These findings provide powerful support for the essence of the dendritic spine abnormalities in the absence of FMRP, now found to be largely consistent with human data across two mouse backgrounds. PMID- 15892135 TI - XBP1 gene polymorphism (-116C/G) and personality. AB - Recently, a polymorphism of the XBP1 gene (-116 C/G) was observed to play a significant role in the development of bipolar mood disorder from the Japanese population. The present study investigated a role of the polymorphism in the development of personality in healthy Japanese volunteers (n = 195). Personality traits were evaluated using NEO Personality Inventory-Revised (NEO PI-R). As a result, a statistical trend for association between the polymorphism (genotype) and the NEO PI-R scores of agreeableness and neuroticism was observed (ANOVA, P = 0.01 and 0.006, respectively). Subjects with the G allele, especially those with G-G genotype, tended to show lower neuroticism and higher agreeableness in the present study. The result is provisional and should be interpreted with caution, partly because the previous study suggested the allele as a risk allele for bipolar disorder. Further studies are required to confirm the results. PMID- 15892136 TI - Bioactive hydroxyapatite coatings on polymer composites for orthopedic implants. AB - Hydroxyapatite [HA, Ca10(PO4)6(OH)2] coatings on polymer composite substrates were investigated for their bioactivity and their physicochemical and mechanical characteristics. HA holds key characteristics for use in orthopedic applications, such as for coating of the femoral stem in a hip replacement device. The plasma spray technique was used to project HA onto a carbon fiber/polyamide 12 composite substrate. The resulting HA coatings exhibited mechanical adhesion as high as 23 MPa, depending on the surface treatment of the composite substrate. The purpose of this investigation was to evaluate the bioactivity of an HA-coated composite substrate. HA- coated samples have been immersed in simulated body fluid (SBF) and maintained within a shaker bath for periods of 1, 7, 14, 21, and 28 days at 37 degrees C. Scanning electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffraction techniques were performed on the samples before and after immersion into SBF. SBF was analyzed using inductively coupled plasma atomic emission spectrometry for element concentration and evaluation of the solution's purity. SBF conditioning led to the deposition of crystalline HA onto the surface of the coatings. The calcium-to-phosphorous ratios of initial HA coating and of newly deposited HA were respectively 1.72 and 1.65, close to the HA theoretical calcium/phosphorous value of 1.67. Results demonstrated that bioactive HA coatings were produced by plasma spraying, because SBF conditioning induced newly formed HA with high crystallinity. Mechanical adhesion of the HA coatings was not significantly affected upon SBF conditioning. PMID- 15892137 TI - Electron density and bonding at inverted carbon atoms: an experimental study of a [1.1.1]propellane derivative. PMID- 15892138 TI - An artificial riboflavin receptor prepared by a template analogue imprinting strategy. PMID- 15892139 TI - Gene expression analysis of peripheral blood leukocytes from discordant sib-pairs with schizophrenia and bipolar disorder reveals points of convergence between genetic and functional genomic approaches. AB - We performed global RNA transcript analysis and comprehensive gene group analysis of peripheral blood leukocyte (PBL) RNA from two groups of matched sib-pairs that were discordant for either schizophrenia (n = 33 sib-pairs) or bipolar disorder (n = 5 sib-pairs). The pairs chosen for these analyses were selected from families with known patterns of genetic linkage (5q for schizophrenia and 6q for bipolar disorder). At the single gene level, we obtained lists of the transcripts with the most significant changes in expression and from these lists determined those with the highest degree of predictive power for classifying subjects according to diagnosis in these samples. At the gene group level, we comprehensively analyzed pairwise expression changes of more than 4,000 functional groups and cytogenetic locations, and present a novel method of displaying these data that we term "cytogenomic" mapping. Verification of selected changes in expression was performed using quantitative real-time RT-PCR. Our results provide compelling evidence for the utility of analyzing PBL RNA for changes in expression in neuropsychiatric disorders. PMID- 15892140 TI - A family study of early-onset obsessive-compulsive disorder. AB - Results from family studies have suggested that obsessive-compulsive disorder (OCD) is a genetically heterogeneous disorder and have emphasized the importance of identifying valid subgroups of patients. The current study focused on early onset OCD probands and examined the recurrence risks of OCD and tics among first degree family members. One hundred six children and adolescents with OCD were recruited from a specialty clinic for OCD and 44 control individuals without OCD were identified by random-digit dialing. These 150 probands and their 465 first degree relatives were assessed by trained interviewers, using standardized semi structured interviews. Diagnoses were assigned according to DSM-IV criteria by two experts blind to the proband's diagnosis, through the best-estimate process. These data were analyzed using chi(2) tests, t-tests, logistic regression, and generalized estimating equations (GEE). Case probands had a mean age of onset of OC symptoms of 6.7 years (SD = 2.8), and high comorbid rates with Tourette syndrome (33%) and chronic tics (13.2%). Compared to control relatives, case relatives had higher age-corrected recurrence risks of OCD (22.7% vs. 0.9%, odds ratio (OR) = 32.5, 95% confidence interval (CI) = 4.5-230.8, P = 0.0005), and chronic tics (11.6% vs. 1.7%, OR = 7.9, 95% CI = 1.9-33.1, P = 0.005). A comorbid diagnosis of tics in the relatives was the best predictor of their diagnosis of OCD (OR = 7.35, 95% CI = 3.79-14.25, P < 0.0001). There was a significant correlation between the ages of onset of OCD in probands and their affected relatives. Childhood onset OCD is a highly familial disorder. Some early-onset cases may represent a valid subgroup, with higher genetic loading and shared vulnerability with chronic tic disorders. PMID- 15892141 TI - One-step synthesis of a platinum(0)-gallium(III) chrysene complex. PMID- 15892142 TI - Enantioselective henry reactions under dual Lewis acid/amine catalysis using chiral amino alcohol ligands. PMID- 15892143 TI - Evaluation of the chromosome 2q37.3 gene CENTG2 as an autism susceptibility gene. AB - Autism is a highly heritable neurodevelopmental syndrome with a complex genetic etiology for which no disease genes have yet been definitively identified. We ascertained three subjects with autism spectrum disorders and chromosome 2q37.3 terminal deletions, and refined the deletion breakpoint regions using polymorphism mapping and fluorescence in situ hybridization (FISH) probes. We then genotyped polymorphic markers downstream from the breakpoint region in a sample of autism affected sibling pair families. Both the chromosomal breakpoints and linkage analyses focused our attention on the gene centaurin gamma-2 (CENTG2), an attractive candidate gene based also on its function and pattern of expression. We therefore assessed CENTG2 for its involvement in autism by (1) screening its exons for variants in 199 autistic and 160 non-autistic individuals, and (2) genotyping and assessing intra-genic polymorphisms for linkage and linkage disequilibrium (LD). The exon screen revealed a Ser --> Gly substitution in one proband, an Arg --> Gly substitution in another, and a number of additional variants unique to the autism families. No unique variants were found in the control subjects. The genotyping produced strong evidence for linkage from two intronic polymorphisms, with a maximum two-point HLOD value of 3.96 and a posterior probability of linkage (PPL) of 51%. These results were contradicted, however, by substantially weaker evidence for linkage from multi point analyses and by no evidence of LD. We conclude, therefore, that 2q37.3 continues to be a region of interest for autism susceptibility, and that CENTG2 is an intriguing candidate gene that merits further scrutiny for its role in autism. PMID- 15892144 TI - Osteoblast interactions with calcium phosphate ceramics modified by coating with type I collagen. AB - Complications associated with the use of autogenous bone in the repair or replacement of tissue lost through injury or disease have driven the search for alternative sources of graft material. Bioceramics containing hydroxyapatite (HA), tricalcium phosphate (TCP), or composites that combine the best properties of both of these materials are among the principal candidates. In this study, we have investigated the in vitro proliferation, morphology, and viability of an immortalized rat osteoblast cell line cultured on HA, TCP, and composites of the two in the ratios 75:25 (H75), 50:50 (H50), and 25:75 (H25) for 28 days. The biocompatibility of each material was examined in the presence and absence of a collagen coating. With the exception of H50, cell proliferation, quantified by carboxyfluorescein fluorescence, was enhanced by collagen coating of all materials for the first 14 days, although at later time points cell numbers were unaffected. It is notable that the collagen coating was least stable on H50, the only material not to show enhancement of cell growth on coating. Confocal laser scanning microscopy confirmed that cell growth was more extensive on coated materials over the first 7-14 days in culture, and the development of cell extensions and bridges across the pores in the materials was observed. Results indicate that collagen coating of calcium phosphate ceramics may also increase their compatibility and osseointegration in vivo. PMID- 15892145 TI - Electronic coupling in mixed-valence dinuclear ferrocenes and cobaltocenes with saturated bridging groups. AB - We have synthesised a series of new dinuclear metallocenes [{M(Cp*)(C5H4)}2X] (Cp* = eta5-pentamethylcyclopentadienyl; M = Fe, Co, X = CMe2, SiMe2, GeMe2; M = Fe, X = Si2Me4). For the neutral dicobalt complexes, magnetic susceptibility measurements reveal intramolecular antiferromagnetic interactions of -21 and -14 cm(-1) for SiMe2- and GeMe2-bridged species, respectively, but negligible interaction for the CMe2-bridged compound. In contrast, intervalence charge transfer (IVCT) data for the mixed-valence monocations of both Fe and Co complexes show electronic coupling to decrease in the order CMe2 > SiMe2 > GeMe2. This suggests that electronic coupling is principally through-space in contrast to results found from previous studies. The IVCT data also show much stronger coupling in the dicobalt species versus their diiron analogues. PMID- 15892146 TI - Synthesis and characterization of thermally robust amidinato group 13 hydride complexes. AB - The reactivity of two sterically bulky amidines, ArNC(R)N(H)Ar (Ar=2,6 diisopropylphenyl; R=H (HFiso); tBu, (HPiso)) towards LiMH4, M=Al or Ga, [AlH3(NMe3)], and [GaH3(quin)] (quin=quinuclidine) has been examined. This has given rise to a variety of very thermally stable aluminum and gallium hydride complexes. The structural motif adopted by the prepared complexes has been found to be dependent upon both the amidinate ligand and the metal involved. The 1:1 reaction of HFiso with LiAlH4 yielded dimeric [{AlH3(mu-Fiso)Li(OEt2)}2]. Amidine HFiso reacts in a 1:1 ratio with [AlH3(NMe3)] to give the unusual hydride bridging dimeric complex, [{AlH2(Fiso)}2], in which the Fiso- ligand is nonchelating. The equivalent reaction with the bulkier amidine, HPiso, yielded a related hydride-bridging complex, [{AlH2(Piso)}2], in which the Piso- ligand is chelating. In contrast, the treatment of [GaH3(quin)] with one equivalent of HFiso afforded the four-coordinate complex [GaH2(quin)(Fiso)], in which the Fiso- ligand acts as a localized monodentate amido-imine ligand. The 2:1 reactions of HFiso with [AlH3(NMe3)] or [GaH3(quin)] gave the monomeric complexes [MH(Fiso)2], which are thermally robust and which exhibit chelating amidinate ligands. In contrast, HPiso did not give 2:1 complexes in its reactions with either of the Group 13 trihydride precursors. For sake of comparison, the reactions of [AlH3(NMe3)] and [GaH3(quin)] with the bulky carbodiimide ArN=C=NAr and the thiourea Ar(H)NC(=S)N(H)Ar were examined. These last reactions afforded the five coordinate thioureido complexes, [MH{N(Ar)C[N(H)(Ar)]S}2], M=Al or Ga. PMID- 15892148 TI - Recent advances in mycotoxin determination in food and feed by hyphenated chromatographic techniques/mass spectrometry. AB - Mycotoxins are fungal toxins produced by molds, which occur universally in food and feed derivatives, and are produced under certain environmental conditions in the field before harvest, post-harvest, during storage, processing, and feeding. Mycotoxin contamination is one of the most relevant and worrisome problem concerning food and feed safety because it can cause a variety of toxic acute and chronic effects in human and animals. In this review we report the use of mass spectrometry in connection with chromatographic techniques for mycotoxin determination by considering separately the most diffuse class of mycotoxins: patulin, aflatoxins, ochratoxin A, zearalenone, trichothecenes, and fumonisins. Although the selectivity of mass spectrometry is unchallenged if compared to common GC and LC detection methods, accuracy, precision, and sensitivity may be extremely variable concerning the different mycotoxins, matrices, and instruments. The sensitivity issue may be a real problem in the case of LC/MS, where the response can be very different for the different ionization techniques (ESI, APCI, APPI). Therefore, when other detection methods (such as fluorescence or UV absorbance) can be used for the quantitative determination, LC/MS appears to be only an outstanding confirmatory technique. In contrast, when the toxins are not volatile and do not bear suitable chromophores or fluorophores, LC/MS appears to be the unique method to perform quantitative and qualitative analyses without requiring any derivatization procedure. The problem of exact quantitative determination in GC/MS and LC/MS methods is particularly important for mycotoxin determination in food, given the high variability of the matrices, and can be solved only by the use of isotopically labeled internal standards or by the use of ionization interfaces able to lower matrix effects and ion suppressions. When the problems linked to inconstant ionization and matrix effects will be solved, only MS detectors will allow to simplify more and more the sample preparation procedures and to avoid clean-up procedures, making feasible low-cost, high throughput determination of mycotoxins in many different food matrices. PMID- 15892149 TI - Sequence variants of the DRD4 gene in autism: further evidence that rare DRD4 7R haplotypes are ADHD specific. AB - A high prevalence of rare dopamine receptor D4 (DRD4) alleles in children diagnosed with attention-deficit hyperactivity disorder (ADHD) has been reported [Grady et al., 2003]. In this prior study, extensive resequencing/haplotype data of the DRD4 locus was used to suggest that population stratification was not the explanation for the high prevalence of rare alleles. In the current study, DNA resequencing/haplotyping was conducted on 136 DRD4 alleles obtained from autism probands, collected from the same geographic population as the prior ADHD probands (Orange County, CA). A number of studies have suggested that the susceptibility genes underlying these two disorders might partially overlap. Rare DRD4 variants were not uncovered in this autism sample beyond that expected by chance. These results suggest strongly that the high prevalence of rare DRD4 alleles in ADHD probands is due to ascertainment of the sample by diagnosis of ADHD. PMID- 15892150 TI - Morbid risk for psychiatric disorder among the relatives of methamphetamine users with and without psychosis. AB - It is not clear why some methamphetamine (MAMP) abusers develop psychotic symptoms, while others use MAMP regularly over long periods and remain unscathed. We tested the hypotheses that those users who develop MAMP-induced psychosis (MIP) have greater familial loading for psychotic disorders than users with no psychosis. Four hundred forty-five MAMP users were recruited from a psychiatric hospital and a detention center in Taipei, and were assessed with the Diagnostic Interview for genetic studies (DIGS-C) and the Family Interview for genetic study (FIGS-C). Morbid risk (MR) for psychiatric disorders in first-degree relatives was compared between those MAMP users with a lifetime diagnosis of MAMP psychosis and those without. The relatives of MAMP users with a lifetime diagnosis of MAMP psychosis had a significantly higher MR for schizophrenia (OR = 5.4, 95% CI: 2.0 14.7, P < 0.001) than the relatives of those probands who never became psychotic. Furthermore, the MR for schizophrenia in the relatives of the subjects with a prolonged MAMP psychosis (MIP-P) was higher than in the relatives of those users with a brief MAMP psychosis (MIP-B) (OR = 2.8, 95% CI: 1.0-8.0, P = 0.042). The greater his or her familial loading for schizophrenia, the more likely a MAMP user is to develop psychosis, and the longer that psychosis is likely to last. PMID- 15892151 TI - Stereoselective functionalization of 2-(1-aminoalkyl)aziridines via lithiation of aziridine-borane complexes. AB - Highly selective functionalization of the aziridine ring of (2S,1'S)-2-(1' aminoalkyl)aziridines 1, through successive formation of aziridine-borane complexes, lithiation, treatment with a variety of electrophiles and final decomplexation is described. The influence of the structure of the starting complexes 2 and of the electrophiles in the stereoselectivity of this process has been studied. Finally, successive double lithiation-electrophile reactions were carried out affording enantiopure 1,2,3,3-tetrasubstituted aziridine-borane complexes with high selectivity. PMID- 15892152 TI - Spatially resolved Raman and UV-visible-NIR spectroscopy on the preparation of supported catalyst bodies: controlling the formation of H2PMo11CoO40 5- inside Al2O3 pellets during impregnation. AB - The physicochemical processes that occur during the preparation of CoMo-Al2O3 hydrodesulfurization catalyst bodies have been investigated. To this end, the distribution of Mo and Co complexes, after impregnation of gamma-Al2O3 pellets with different CoMoP solutions (i.e., solutions containing Co, Mo, and phosphate), was monitored by Raman and UV-visible-NIR microspectroscopy. From the speciation of the different complexes over the catalyst bodies, insight was obtained into the interaction of the different components in the impregnation solution with the Al2O3 surface. It is shown that, after impregnation with a solution containing H2PMo11CoO40(5-), the reaction of phosphate with the Al2O3 leads to the disintegration of this complex. The consecutive independent transport of Co2+ complexes (fast) and Mo6+ complexes (slow) through the pores of the Al2O3 is envisaged. By the addition of extra phosphate and citrate to the impregnation solution, the formation of the desired heteropolyanion can be achieved inside the pellets. Ultimately, the H2PMo11CoO40(5-) distribution could be controlled by varying the aging time applied after impregnation. The power of a combination of spatially resolved spectroscopic techniques to monitor the preparation of supported catalyst bodies is illustrated. PMID- 15892153 TI - Quality of life assessment in patients with chronic anal fissure after lateral internal sphincterotomy. AB - BACKGROUND: The aim of this study was to compare quality of life of patients with chronic anal fissure before and after open lateral internal sphincterotomy. METHODS: A prospective study was undertaken of 108 consecutive patients with a history of chronic anal fissure who underwent lateral internal sphincterotomy. Quality of life was measured before and 6 months after operation with the Short Form 36 (SF-36) Health Survey. RESULTS: Quality of life improved significantly in six of the eight scales of the SF-36 questionnaire: physical functioning, role physical, bodily pain, energy, social functioning and mental health. There were no significant differences between the 70 patients who had no change in continence after operation and the 38 patients with continence disturbances after sphincterotomy. However, there were significant improvements in four scales in patients without changes in continence compared with improvements in only two scales in those with continence disturbances. CONCLUSION: Patients with chronic anal fissure showed an improvement in quality of life 6 months after internal lateral sphincterotomy. Patients with postoperative continence disturbances showed improvement in fewer scales of the SF-36 questionnaire than those without changes in continence. PMID- 15892154 TI - Risk of tumour seeding after percutaneous radiofrequency ablation for hepatocellular carcinoma. AB - BACKGROUND: A recent small study reported a high rate of neoplastic seeding after cooled-tip radiofrequency ablation (RFA) for hepatocellular carcinoma (HCC) in patients who had undergone previous needle biopsy. Tumour seeding was associated with subcapsular tumour location, poorly differentiated tumours and a high alpha fetoprotein (AFP) level. The aim of the present study was to determine the rate of neoplastic seeding after RFA in a large series of unselected patients with HCC who had a long follow-up. METHODS: A total of 1314 patients with 2542 nodules were treated in three centres. Median follow-up was 37 months. Needle biopsy had been performed before RFA in 241 patients (18.3 per cent). The influence of subcapsular location, high AFP level and previous biopsy on risk of tumour seeding was assessed. RESULTS: Neoplastic seeding was identified in 12 patients (0.9 per cent); the rate was comparable at the three centres (0.9, 0.7 and 1.4 per cent). Only previous biopsy was significantly associated with tumour seeding (P = 0.004). CONCLUSION: RFA with a cooled-tip needle was associated with a low risk of neoplastic seeding, even in unselected patients. The use of biopsy before RFA is to be discouraged, particularly when liver transplantation is a possibility at a later date. PMID- 15892155 TI - Population-based study of short- and long-term survival from colorectal cancer in Denmark, 1977-1999. AB - BACKGROUND: Survival rates for patients with colorectal cancer have been lower in Denmark than in other European countries. The aim of this study was to examine temporal trends in relative survival from colorectal cancer between 1977 and 1999. METHODS: All patients diagnosed with colorectal cancer between 1977 and 1999 were identified using the nationwide population-based Danish Cancer Registry. Patients were linked with the Danish Central Population Registry to obtain data on survival to December 2002, and to select ten population controls per patient. RESULTS: A total of 69 562 patients with colorectal cancer were identified, of whom 49.2 per cent were men. Six-month relative survival after diagnosis increased from 69.7 per cent in 1977-1982 to 77.7 per cent in 1995 1999. Five-year relative survival rates increased from 37.8 to 46.8 per cent respectively. Women had slightly higher 5-year relative survival than men throughout the study period. Rectal tumours were associated with better survival than colonic tumours until 2 years after diagnosis, after which tumour location had no impact on survival. CONCLUSION: Relative survival of patients with colorectal cancer has improved in Denmark between 1977 and 1999, most probably reflecting better management of the disease. PMID- 15892156 TI - Surgical outcome of solitary colorectal metastasis to the hepatic caudate lobe. PMID- 15892157 TI - Prognostic significance of alarm symptoms in patients with gastric cancer. AB - BACKGROUND: The aim of this study was to determine the incidence and spectrum of alarm symptoms in patients with newly diagnosed gastric cancer, and to examine the relationship between symptoms and outcome. METHODS: Three hundred consecutive patients with gastric adenocarcinoma were studied prospectively. The outcomes of 40 patients (13.3 per cent) without alarm symptoms (21 men; median age 69 years) were compared with those of the 260 patients (86.7 per cent) with alarm symptoms (175 men; median age 72 years). RESULTS: It was possible to perform an R0 gastrectomy more often in patients without alarm symptoms (21 patients; 52 per cent) than in those with alarm symptoms (71 patients; 27.3 per cent) (chi(2) = 10.35, 1 d.f., P = 0.001). The cumulative survival rate at 5 years was 38 per cent for patients without alarm symptoms versus 15.0 per cent for those with alarm symptoms (chi(2) = 10.18, 1 d.f., P = 0.001). In a multivariate analysis, distant metastasis (hazard ratio (HR) 2.73 (95 per cent confidence interval (c.i.) 2.04 to 3.66); P < 0.001), overall stage of cancer (HR 1.83 (95 per cent c.i. 1.53 to 2.19); P < 0.001) and persistent vomiting at diagnosis (HR 1.66 (95 per cent c.i. 1.26 to 2.18); P < 0.001) were independently associated with length of survival. CONCLUSION: Alarm symptoms are absent in a significant minority of patients with gastric cancer at diagnosis; these patients stand a better chance of curative surgery and long-term survival than those with alarm symptoms. PMID- 15892158 TI - Minimally invasive transthoracotomy-transphrenotomy for concurrent hepatic and pulmonary hydatid disease. AB - BACKGROUND: Combined liver and lung hydatid cysts are rare, but pose a challenge in terms of accessibility. The objective of the study was to find an alternative approach to conventional two-stage posterolateral thoracotomy and laparotomy or single-stage extensive thoracolaparotomy. METHODS: Twenty-five patients with right lung and liver hydatid disease underwent single-stage anterior minithoracotomy and phrenotomy. Primary diagnostic tools were chest radiography, ultrasonography and serology. The preferred mode of management of hydatid cysts was enucleation and partial or total capitonnage. RESULTS: Thirty-six (13.5 per cent) of 267 patients had concurrent hepatic and pulmonary hydatid cysts. Among the 25 patients who had anterior minithoracotomy and phrenotomy the male : female ratio was 2 : 1. Mean operating time was 75 min. Morbidity was negligible and postoperative recovery was prompt. All of the patients survived. The mean hospital stay was 5.2 days. Overall observations were encouraging. CONCLUSION: This minimally invasive approach is associated with less morbidity and better cosmesis than conventional procedures. It represents an excellent alternative to other procedures in selected patients. PMID- 15892159 TI - Detection of sentinel nodes and micrometastases using radioisotope navigation and immunohistochemistry in patients with gastric cancer. AB - BACKGROUND: Patients with early gastric cancer may be treated by minimally invasive surgery. This study investigated the value of sentinel node (SN) navigation surgery, including detection of micrometastases, in patients with clinical (c) T1 and T2 gastric cancer. METHODS: The day before surgery (99m)Tc radiolabelled tin colloid was injected submucosally near the tumour. After resecting the stomach, radioisotope uptake in all dissected lymph nodes was measured during and after surgery. Micrometastasis was detected immunohistochemically using an anticytokeratin antibody. RESULTS: SNs were identified in 99 of 104 patients. The rate of identification of SNs in patients with cT1 and cT2 tumours, excluding three technical failures, was 99 and 95 per cent respectively. Lymph node metastases and/or micrometastases were found in 28 patients (15 cT1 and 13 cT2). In the 15 patients with cT1 tumours, at least one SN contained metastasis and/or micrometastasis. For cT1 tumours, the sensitivity and accuracy of detecting SNs were both 100 per cent. Six patients with cT2 tumours had false-negative results. CONCLUSION: SN navigation surgery appears to be clinically useful only for cT1 tumours. Based on SN results, the extent of lymphadenectomy may be reduced in patients with early gastric cancer. PMID- 15892160 TI - Durability of botulinum toxin injection for axillary hyperhidrosis. PMID- 15892161 TI - Experimental bile duct protection by intraductal cooling during radiofrequency ablation. AB - BACKGROUND: The use of radiofrequency ablation (RFA) for liver tumours is limited by the proximity of large bile ducts to the targeted lesion. The aim of this randomized study was to evaluate intraductal cooling as a mean of protecting the bile ducts during RFA. METHODS: Twelve pigs underwent RFA adjacent to the right bile duct. After placement of an intraductal cooling catheter and a RFA probe, pigs were randomized to cooling or no cooling. Intraductal temperature was measured in all animals. The bile ducts were assessed by magnetic resonance imaging (MRI) and cholangiography 1 and 28 days after the procedure. RESULTS: Intraductal cooling abolished the increase of intraductal temperature seen in the absence of cooling. Concurrent cholangiography and MRI showed a biliary lesion in one of six pigs subjected to intraductal cooling and in five of six without cooling (P = 0.040). The biliary injuries were barely visible by MRI on day 1 but were clearly visible on day 28. CONCLUSION: Intraductal cooling can prevent biliary injury induced by RFA. The exact parameters for intraductal cooling require further investigation to establish the best compromise between bile duct protection and complete ablation of surrounding tissue. PMID- 15892162 TI - Proteomic approach to blossom-end rot in tomato fruits (Lycopersicon esculentum M.): antioxidant enzymes and the pentose phosphate pathway. AB - Blossom-end rot (BER) is a physiopathy that affects tomato fruits causing disorganisation, cell breakage and darkening of the tissues. In this study we describe a tomato fruit protein extraction protocol that includes polyvinyl polypyrrolidone, ascorbic acid and protease inhibitors to promote depletion of phenolics and to avoid protein degradation. The temperature-induced phase separation of plant extracts with nonionic detergent Triton X-114 favours the solubilisation of partially-hydrophobic species in the low-detergent upper phase, making them suitable for further analysis using two-dimensional gel electrophoresis. The analysis of two-dimensional images revealed differences in number and expression levels of several proteins from the control and BER affected tomato fruits. Although the appearance of BER in tomato is primarily attributed to a lack of calcium supply to fruits, very little is known about the molecular and biochemical mechanisms involved. The identification of differential proteins from affected fruits with matrix-assisted laser desorption/ionisation time of flight and peptide mass fingerprinting analysis revealed the induction of proteins participating in antioxidant processes (ascorbate-glutathione cycle) and the pentose phosphate pathway. We suggest that these two biochemical pathways, acting as reactive oxygen species scavengers in BER-affected fruits, restrain the spread of the blackening to the whole fruit. PMID- 15892163 TI - Differential protein expression in anatomical zones of the prostate. AB - The prostate has three anatomical zones: the peripheral (PZ), the transition (TZ), and the central (CZ) zone. It is proposed that the CZ may be of mesodermal origin, whereas the other two are of endodermal origin. Proteome patterns in the zones were characterized to test for differences. Cells were scraped from macroscopically normal areas of PZ, TZ, and CZ in radical prostatectomy specimens. After exclusion of samples with cancer or prostatic intraepithelial neoplasia, 18 cases remained for analysis. Cells were collected in a medium with protease inhibitors, and the protein material was prepared for two-dimensional gel electrophoresis. The proteins in spots that differed quantitatively between regions were identified via mass spectrometric fingerprinting of tryptic fragments and selected tandem mass spectrometry sequence analysis. Ten proteins with significant zonal differential expression were identified, eight with underexpression in the CZ versus the PZ and the TZ (arginase II, ATP synthase, cytokeratin 8, lamin A/C, peroxiredoxin 4, protein disulfide isomerase A3, tropomyosin, and vimentin), and two with overexpression in the CZ (peroxiredoxin 2 and creatine kinase B). The PZ and TZ, although differing in terms of incidence of cancer and hyperplasia, have epithelium with highly similar major protein expression profiles. However, the protein profile of the CZ differs from that of the other regions, suggesting functional differences. PMID- 15892164 TI - Mass spectrometry-based analytical tools for the molecular protein characterization of human plasma lipoproteins. AB - Lipoproteins are a heterogeneous population of blood plasma particles composed of apolipoproteins and lipids. Lipoproteins transport exogenous and endogenous triglycerides and cholesterol from sites of absorption and formation to sites of storage and usage. Three major classes of lipoproteins are distinguished according to their density: high-density (HDL), low-density (LDL) and very low density lipoproteins (VLDL). While HDLs contain mainly apolipoproteins of lower molecular weight, the two other classes contain apolipoprotein B and apolipoprotein (a) together with triglycerides and cholesterol. HDL concentrations were found to be inversely related to coronary heart disease and LDL/VLDL concentrations directly related. Although many studies have been published in this area, few have concentrated on the exact protein composition of lipoprotein particles. Lipoproteins were separated by density gradient ultracentrifugation into different subclasses. Native gel electrophoresis revealed different gel migration behaviour of the particles, with less dense particles having higher apparent hydrodynamic radii than denser particles. Apolipoprotein composition profiles were measured by matrix-assisted laser desorption/ionization-mass spectrometry on a macromizer instrument, equipped with the recently introduced cryodetector technology, and revealed differences in apolipoprotein composition between HDL subclasses. By combining these profiles with protein identifications from native and denaturing polyacrylamide gels by liquid chromatography-tandem mass spectrometry, we characterized comprehensively the exact protein composition of different lipoprotein particles. We concluded that the differential display of protein weight information acquired by macromizer mass spectrometry is an excellent tool for revealing structural variations of different lipoprotein particles, and hence the foundation is laid for the screening of cardiovascular disease risk factors associated with lipoproteins. PMID- 15892165 TI - First evidence supporting a potential role for the BMP/SMAD pathway in the progression of oestrogen receptor-positive breast cancer. AB - Oestrogen receptor expression is generally a sign of better tumour differentiation and comparatively good clinical outcome in invasive breast cancer. However, oestrogen receptor-positive, poorly differentiated carcinomas with a poor clinical outcome exist. The underlying genetic mechanisms and the genes involved remain obscure, even though chromosome 7p gains seem to be associated with these uncommon tumours. In this study, we compared two subsets of oestrogen receptor-positive breast cancers, which differed in tumour grade, cytogenetic instability, and tumour proliferation, for their differential gene expression in order to identify proteins involved in the progression of oestrogen receptor-positive breast cancers. We were able to show by means of subtractive suppression hybridization, real-time reverse transcriptase PCR, and tissue microarray analysis that expression of the bone morphogenetic protein receptor IB (BMPR-IB) is a major hallmark of the progression and dedifferentiation of breast cancer. Strong expression of BMPR-IB was associated with high tumour grade, high tumour proliferation, cytogenetic instability, and a poor prognosis in oestrogen receptor-positive carcinomas. Western blot analysis revealed that downstream signalling of this receptor is mainly mediated via phosphorylation of SMAD 1 in oestrogen receptor-positive breast cancer. Even though BMPR-IB was expressed in oestrogen receptor-positive and -negative breast cancers, an impact on tumour grade, proliferation, and cytogenetic instability, as parameters of tumour progression, could only be demonstrated in oestrogen receptor-positive carcinomas. This pro-proliferative effect was complemented by significant anti apoptotic activity, indicated by XIAP and IAP-2 expression in BMPR-IB-positive carcinomas. These results show that the BMP/SMAD pathway is activated in breast cancer and may contribute to breast cancer progression and dedifferentiation in oestrogen receptor-positive breast cancer. The definition of this pathway characterizes a new potential target in the molecular treatment of invasive breast cancer. PMID- 15892166 TI - Cardiac troponin I predicts outcome after ruptured abdominal aortic aneurysm repair. AB - BACKGROUND: Cardiac troponin I (cTnI) is a highly sensitive and specific marker for myocardial injury that predicts mortality in patients with acute coronary syndromes. This study examined the relationship between perioperative cTnI levels and clinical outcome in patients with ruptured abdominal aortic aneurysm (AAA). METHODS: Consecutive patients who underwent operative repair of a ruptured AAA over a 22-month interval and survived for more than 24 h were entered into a prospective observational cohort study. Levels of cTnI were measured immediately before, and at 24 and 48 h after surgery, and related to clinical outcome. RESULTS: Of 62 patients who underwent attempted operative repair of ruptured AAA, 50 (81 per cent) survived for more than 24 h and were included in this study. Twenty-three (46 per cent) of the 50 had a detectable cTnI level at one or more time points during the first 48 h. Of these, 11 patients had clinical or electrocardiographic evidence of an acute cardiac event and 12 did not; five patients in each of these two groups died. Of 27 patients with no increase in cTnI in the first 48 h, only three died (P = 0.031 and P = 0.043 respectively, relative to the groups with detectable cTnI). CONCLUSION: Approximately half of patients who survived repair of ruptured AAA for more than 24 h sustained a detectable myocardial injury within the first 48 h. A perioperative increase in the level of cTnI, with or without clinically apparent cardiac dysfunction, was associated with postoperative death. PMID- 15892167 TI - Identification of fatty acid binding proteins as markers associated with the initiation and/or progression of renal cell carcinoma. AB - Renal cell carcinoma (RCC) representing the most common neoplasia of the kidney in Western countries is a histologic diverse disease with an often unpredictable course. The prognosis of RCC is worsened with the onset of metastasis, and the therapies currently available are of limited success for the treatment of metastatic RCC. Although gene expression analyses and other methods are promising tools clarifying and standardizing the pathological classification of RCC, novel innovative molecular markers for the diagnosis, prognosis, and for the monitoring of this disease during therapy as well as potential therapeutic targets are urgently needed. Using proteome-based strategies, a number of RCC-associated markers either over-expressed or down-regulated in tumor lesions in comparison to the normal epithelium have been identified which have been implicated in tumorigenesis, but never linked to the initiation and/or progression of RCC. These include members of the fatty acid binding protein family, which have the potential to serve as diagnostic or prognostic markers for the screening of RCC patients. PMID- 15892168 TI - Method optimisation for peptide profiling of microdissected breast carcinoma tissue by matrix-assisted laser desorption/ionisation-time of flight and matrix assisted laser desorption/ionisation-time of flight/time of flight-mass spectrometry. AB - Appropriate methods for the analysis of microdissected solid tumour tissues by matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF MS) are not yet well established. Optimisation of sample preparation was performed first on undissected tissue slices, representing approximately 200 000 cells, which were solubilised either in urea containing buffer, trifluoroethanol/NH4HCO3, 0.1% sodium dodecyl sulphate (SDS) or in 0.1% RapiGest solution, then trypsin digested and analysed by MALDI-TOF MS. Solubilisation in 0.1% SDS resulted in detection of the highest number of sample specific peak signals. Interestingly, there was little overlap in detectable peaks using the different buffers, implying that they can be used complementarily to each other. Additionally, we fractionated tryptic digests on a monolithic high-performance liquid chromatography column. Fractionation of tryptic digest from whole tissue sections resulted in a four-fold increase in the total number of peaks detected. To prove this principle, we used 0.1% SDS to generate peptide patterns from 2000 microdissected tumour and stromal cells from five different breast carcinoma tumours. The tumour and stroma specific peaks could be detected upon comparison of the peptide profiles. Identification of differentially expressed peaks by MALDI-TOF/TOF MS was performed on fractionated tryptic digests derived from a whole tissue slice. In conclusion, we describe a method that is suitable for direct peptide profiling on small amounts of microdissected cells obtained from breast cancer tissues. PMID- 15892169 TI - Rates of abuse of tramadol remain unchanged with the introduction of new branded and generic products: results of an abuse monitoring system, 1994-2004. AB - PURPOSE: The analgesic Tramadol HCl (Ultram) was approved in 1994 as a non scheduled drug under the CSA provided that a novel risk-management program would be developed by an Independent Steering Committee (ISC). The risk-management program began in 1995 with the launch of Ultram, and has been modified over the past decade to accommodate Ultracet (Ultram and acetaminophen) in 2001 and generic tramadol in 2002. This provided a unique opportunity to study the potential changes in abuse as the generic and combination products became available. METHODS: To proactively detect cases of abuse and diversion, the ISC developed a comprehensive questionnaire which was completed quarterly by an extensive network of drug abuse experts (n = 309) and police agencies (n = 100) who were asked to indicate how many diversion cases involving Ultram, Ultracet, and generic tramadol were identified during the preceding 3 months and what were the ten most commonly diverted drugs in their catchment area during that period. RESULTS AND CONCLUSIONS: The data generated demonstrate that the abuse of tramadol remained very low despite new branded and generic formulations. Contrary to the hypothesis that cheaper generic drugs would lead to higher rates of abuse, we found no increase in abuse with the introduction of generic tramadol. Ultracet abuse rates, unlike those found with other widely used hydrocodone and oxycodone combination products, have been even lower than that observed for tramadol. Since the FDA has now mandated that proactive risk-management plans be implemented for new drugs, the tramadol risk-management plan may be useful as a prototypic model which can be modified to accommodate other drugs with abuse potential. PMID- 15892170 TI - The negative effect of red tape on research. AB - PURPOSES: To describe the kind of the difficulties encountered when seeking research governance approval for a nationwide public health and genetic study-the Drug-Induced Arrhythmia Risk Evaluation study-in England. METHODS: Description of the processes followed when seeking research governance approval for the Drug Induced Arrhythmia Risk Evaluation study-a case control study with annual follow up of cases and controls over 5 years, set in the English National Health Service (NHS). RESULTS: The authors describe wide variations in NHS research governance approval procedures in England. CONCLUSION: NHS research governance procedures in England are impeding the process of epidemiological studies; there is the need for a centralised NHS R&D approval of studies, which is analogous to MREC for ethical approval. PMID- 15892171 TI - PU.1 protein expression has a positive linear association with protein expression of germinal centre B cell genes including BCL-6, CD10, CD20 and CD22: identification of PU.1 putative binding sites in the BCL-6 promotor. AB - The transcription factor PU.1 has been shown to be crucial for the early stages of B cell development but its function at later stages of B cell development is less well known. We observed previously that PU.1 is expressed uniformly throughout the mature pre-plasma cell B cell population, the only exception being a subpopulation of germinal centre (GC) cells which showed exceptionally high expression of PU.1. This suggested that PU.1 may also have a role in GC B cell biology. To test this hypothesis and to screen for possible genes regulated by PU.1, we first evaluated semi-quantitatively the possible co-expression of PU.1 with proteins known to be upregulated or downregulated during GC B cell development. Normal lymphoid tissues and 255 B cell non-Hodgkin lymphomas of putative GC B cell origin were evaluated. PU.1 expression was positively associated with CD10 (p < 0.0001), CD20 (p = 0.043), CD22 (p = 0.005), CD79a (p = 0.024) and Bcl-6 (p < 0.0001) and negatively associated with cytoplasmic immunoglobulin light-chain expression (p = 0.036) in diffuse large B cell lymphoma. Identical or nearly identical associations were found in follicular lymphoma. Since CD20 is known to be partly regulated by PU.1 and putative PU.1 binding sites have been described in the regulatory regions of the CD22, CD79a and CD10 genes, we looked for putative PU.1 binding sites in the BCL6 promotor. Four such putative PU.1 binding sites were identified. Further analysis by gel shift electromobility essay showed that PU.1 protein binds to three of the four putative binding sites in the BCL6 promotor. PU.1 and Bcl-6 were also found to be upregulated in centroblasts in the normal GC, but jointly downregulated in a subpopulation of centrocytes. Our findings support the contention that PU.1 may also have an important role in GC B cell development. PMID- 15892172 TI - Amplification and overexpression of c-erbB-2, epidermal growth factor receptor, and c-met in biliary tract cancers. AB - Carcinomas of the biliary tract have a poor prognosis. It is important to understand the molecular genetic characteristics of these tumours in order to employ newer effective treatments and to improve patient prognosis. There is increasing evidence that overexpression of tyrosine kinase growth factor receptors such as ErbB-2, epidermal growth factor receptor (EGFR), and Met may play important roles in the development of biliary tract carcinomas. The aim of this study was to assess the potential for novel chemotherapies targeting these receptors. Overexpression of the tyrosine kinase receptor proteins was examined by immunohistochemistry in 221 biliary tract carcinomas, of which 28 were from the intrahepatic bile duct, 78 from the extrahepatic bile duct, 89 from the gall bladder, and 26 from the ampulla of Vater. Positively stained tumours were further examined for gene amplification by fluorescence in situ hybridization. Overexpression of ErbB-2 was found in 15.7%, 11.5%, and 5.1% of carcinomas of the gall bladder, ampulla of Vater, and extrahepatic bile duct, respectively, and gene amplification was present in 79% of these. Overexpression of EGFR was found in 8.1% of tumours with no predominant location and was also associated with gene amplification with high frequency (77%). Met overexpression, most frequent in intrahepatic bile duct carcinomas (21.4%), was not associated with gene amplification. It is proposed that the new adjuvant chemotherapies could be directed to carcinomas of the biliary tract in which ErbB-2 and EGFR are overexpressed. PMID- 15892173 TI - Proteomic analysis of anti-Francisella tularensis LVS antibody response in murine model of tularemia. AB - Francisella tularensis live vaccine strain infection of mice has been established as an experimental model of tularemia that is suitable for studies of immune mechanisms against the intracellular pathogen. In this study, the model was used to explore immunogenic repertoire of F. tularensis with the aim of identifying new molecules able to activate the host immune system, potential bacterial markers with vaccine, and diagnostic applications. Immunoproteomic approach based on the combination of two-dimensional gel electrophoresis, immunoblotting, and mass spectrometry was applied. Globally, 36 different proteins were identified, which strongly reacted with sera from experimentally infected mice, including several putative virulence markers of intracellular pathogens as nucleoside diphosphate kinase, isocitrate dehydrogenase, RNA-binding protein Hfq, and molecular chaperone ClpB. Of them, 27 proteins are described for the first time as immunorelevant Francisella proteins. When comparing murine immunoproteome of F. tularensis with our previous data from human patients, 25 of the total of 50 identified murine sera immunoreactive spots were recognized by human sera collected from patients suffering from tularemia, as well. Immune sera from two Lps gene congenic strains of mice, C3H/HeN (Lpsn) and C3H/HeJ (Lpsd), represented murine immunoproteome in this study. The spectrum of immunoreactive spots detected by two-dimensional immunoblotting varied throughout the course of infection depending on murine strain. Nevertheless, the antibody patterns of the two strains showed significant homogeneity in being directed against almost identical subset of antigens. PMID- 15892174 TI - Structure elucidation of aplidine metabolites formed in vitro by human liver microsomes using triple quadrupole mass spectrometry. AB - The cyclic depsipeptide aplidine is a new anti-cancer drug of marine origin. Four metabolites of this compound were found after incubation with pooled human microsomes using gradient high-performance liquid chromatography with ultraviolet detection. After chromatographic isolation, the metabolites have been identified using nano-electrospray triple quadrupole mass spectrometry. A highly specific sodium-ion interaction with the cyclic structure opens the depsipeptide ring, and cleavage of the amino acid residues gives sequence information when activated by collision-induced dissociation in the second quadrupole. The aplidine molecule could undergo the following metabolic reactions: hydroxylation at the isopropyl group (metabolites apli-h 1 and apli-h 2); C-dealkylation at the N(Me)-leucine group (metabolite apli-da); hydroxylation at the isopropyl group and C dealkylation at the N(Me)-leucine group (metabolite apli-da/h), and C demethylation at the threonine group (metabolite apli-dm). The identification of these metabolites formed in vitro may greatly aid the elucidation of the metabolic pathways of aplidine in humans. PMID- 15892175 TI - Changes in isotretinoin prescribing before and after implementation of the System to Manage Accutane Related Teratogenicity (SMART) risk management program. AB - PURPOSE: To assess changes in isotretinoin prescribing following the implementation of the System to Manage Accutane Related Teratogenicity (SMART) risk management program. METHODS: Using nationally representative commercial data resources on prescription drug dispensing patterns, surveys of office-based physician practices, and a large, claims database from a pharmacy benefits manager (PBM), we examined the total number of isotretinoin prescriptions (new and refill), prescriber speciality, and patient characteristics (age, gender, severity of acne indication) in the year before (April 2001-March 2002) and the year following (April 2002-March 2003) implementation of the SMART program. RESULTS: In the 12-months prior to SMART, 1 508 000 prescriptions were dispensed for isotretinoin, declining approximately 23% to 1 160 000 prescriptions in the year following SMART. There was little or no change in prescriber specialty, severity of acne, and patient age and gender. CONCLUSION: SMART may have lead to a decrease in isotretinoin prescriptions. Further research is needed to determine whether the reduced number of isotretinoin prescriptions reflects appropriate use or inhibited use resulting in loss of access to the product's benefits. PMID- 15892177 TI - Measurement of pulmonary surfactant disaturated-phosphatidylcholine synthesis in human infants using deuterium incorporation from body water. AB - The aim of the study was to determine surfactant palmitate disaturated phosphatidylcholine (DSPC-PA) synthesis in vivo in humans by the incorporation of deuterium from total body water into DSPC-PA under steady state condition. We studied three newborns and one infant (body weight (BW) 4.6 +/- 2.9 kg, gestational age 37.5 +/- 2 weeks, age 9 +/- 9 days) and four preterm newborns (BW 1.3 +/- 0.6 kg, gestational age 30.3 +/- 2.5 weeks, postnatal age 8.8 +/- 9.2 h). All infants were mechanically ventilated during the study and the four preterm infants received exogenous surfactant at the start of the study. We administered 0.44 g (2)H(2)O/kg BW as a bolus intravenously, followed by 0.0125 g (2)H(2)O/kg BW every 6 h to maintain deuterium enrichment at plateau over 2 days. Urine samples and tracheal aspirates (TA) were obtained prior to dosing and every 6 h thereafter. Isotopic enrichment curves of DSPC-PA from sequential TA and urine deuterium enrichments were analyzed by Gas Chromatography-Isotope Ratio-Mass Spectrometry (GC-IRMS) and normalized for Vienna Standard Mean Ocean Water. Enrichment data were used to measure DSPC-PA fractional synthesis rate (FSR) from the linear portion of the DSPC-PA enrichment rise over time, relative to plateau enrichment of urine deuterium. Secretion time (ST) was defined as the time lag between the start of the study and the appearance of DSPC-PA deuterium enrichment in TA. Data were given as mean +/- SD. All study infants reached deuterium-steady state in urine. DSPC-PA FSR was 6.5 +/- 2.8%/day (range 2.6-10.2). FSR for infants who did not receive exogenous surfactant was 5.7 +/- 3.5%/day (range 2.6 9.9%/day) and 7.3 +/- 2.1%/day (range 5.1-10.2%/day) in the preterms, whereas DSPC-PA ST was 10 +/- 10 h and 31 +/- 10 h respectively. Surfactant DSPC-PA synthesis can be measured in humans by the incorporation of deuterium from body water. This study is a simpler and less invasive method compared to previously published methods on surfactant kinetics by means of stable isotopes. PMID- 15892178 TI - Measurement of bile acid CoA esters by high-performance liquid chromatography electrospray ionisation tandem mass spectrometry (HPLC-ESI-MS/MS). AB - The novel and rapid assay presented here combines high-performance liquid chromatography and electrospray ionisation tandem mass spectrometry (HPLC-ESI MS/MS) to directly measure and quantify the CoA esters of 3alpha,7alpha,12alpha trihydroxy- and 3alpha,7alpha-dihydroxy-5beta-cholestan-26-oic acid (THCA and DHCA). The latter are converted inside peroxisomes to the primary bile acids, cholic and chenodeoxycholic acids, respectively. Prior to MS/MS, esters were separated by reversed-phase HPLC on a C(18) column using an isocratic mobile phase (acetonitrile/water/2-propanol) and subsequently detected by multiple reaction monitoring. For quantification, the CoA ester of deuterium-labelled 3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oic acid (d(4)-CA) was used as internal standard. To complete an assay took less than 8 min. To verify the validity of the assay, the effect of peroxisomal proteins on the efficacy of extraction of the CoA esters was tested. To this end, variable amounts of the CoA esters were spiked with a fixed amount of either intact peroxisomes or peroxisomal matrix proteins and then extracted using a solid-phase extraction system. The CoA esters could be reproducibly recovered in the range of 0.1-4 micromol l(-1) (linear correlation coefficient R(2) > 0.99), with a detection limit of 0.1 micromol l(-1). In summary, electrospray ionization tandem mass spectrometry combined with HPLC as described here proved to be a rapid and versatile technique for the determination of bile acid CoA esters in a mixture with peroxisomal proteins. This suggests this technique to become a valuable tool in studies dealing with the multi-step biosynthesis of bile acids and its disturbances in disorders like the Zellweger syndrome. PMID- 15892181 TI - Production of the tubulin destabilizer disorazol in Sorangium cellulosum: biosynthetic machinery and regulatory genes. AB - Myxobacteria show a high potential for the production of natural compounds that exhibit a wide variety of antibiotic, antifungal, and cytotoxic activities. The genus Sorangium is of special biotechnological interest because it produces almost half of the secondary metabolites isolated from these microorganisms. We describe a transposon-mutagenesis approach to identifying the disorazol biosynthetic gene cluster in Sorangium cellulosum So ce12, a producer of multiple natural products. In addition to the highly effective disorazol-type tubulin destabilizers, S. cellulosum So ce12 produces sorangicins, potent eubacterial RNA polymerase inhibitors, bactericidal sorangiolides, and the antifungal chivosazoles. To obtain a transposon library of sufficient size suitable for the identification of the presumed biosynthetic gene clusters, an efficient transformation method was developed. We present here the first electroporation protocol for a strain of the genus Sorangium. The transposon library was screened for disorazol-negative mutants. This approach led to the identification of the corresponding trans-acyltransferase core biosynthetic gene cluster together with a region in the chromosome that is likely to be involved in disorazol biosynthesis. A third region in the genome harbors another gene that is presumed to be involved in the regulation of disorazol production. A detailed analysis of the biosynthetic and regulatory genes is presented in this paper. PMID- 15892182 TI - Antioxidants and an inhibitor of advanced glycation ameliorate death of retinal microvascular cells in diabetic retinopathy. AB - BACKGROUND: Pericyte ghosts and acellular capillaries are well known as early histological changes resulting from diabetic retinopathy. These histological changes mean that the cell death of retinal microvessels has accelerated. It was reported that apoptosis of retinal microvascular cells (RMCs) was increased in diabetic patients. Therefore, we investigated apoptosis of RMCs in Goto-Kakizaki (GK) rats, a type 2 diabetic model, and involvement with antioxidants (a combination of vitamins C and E) or a novel inhibitor of advanced glycation, OPB 9195. METHODS: GK rats were treated with the antioxidants combination or OPB-9195 for 36 weeks. We obtained isolated preparations of the vascular network from their retinas by trypsin digestion. Apoptosis of retinal vascular cells was detected with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. RESULTS: We found that apoptosis of RMCs was increased in the diabetic GK rats. Furthermore, a combination of vitamins C and E and an advanced glycation end-products inhibitor mostly inhibited this increased apoptosis. CONCLUSIONS: We concluded that apoptosis of RMCs was a good marker that indicates the progression of diabetic retinopathy in GK rats. Both oxidative stress and the accumulation of advanced glycation end-products appears to promote the apoptosis of retinal microvascular cells, and antioxidants or advanced glycation end-products inhibitors might ameliorate diabetic retinopathy. PMID- 15892183 TI - Electrospray ionization mass and tandem mass spectra of a series of N pyrazolylmethyl and N-triazolylmethyl N-phenylpiperazines: new dopaminergic ligands with potential antipsychotic properties. AB - Recently, two analogous series of N-pyrazolylmethyl and N-triazolylmethyl N phenylpiperazines have been prepared and found to be potential antipsychotic drugs acting as new selective ligands of the dopamine D2 receptor. Herein we report a systematic study of their high-resolution electrospray ionization mass and tandem mass spectra in which the main dissociation routes of their protonated molecules are determined and rationalized. The ESI-MS/MS data is very characteristic for both series allowing straightforward isomeric differentiation. A single and dominant fragment ion for the pyrazole series and four major fragment ions for the triazole series are useful for selective reaction MS monitoring of these potential drugs in biological fluids. PMID- 15892184 TI - Molecular aptamers for real-time protein-protein interaction study. AB - Protein-protein interactions play critical roles in cellular functions, but current techniques for real-time study of these interactions are limited. We report the real-time monitoring of protein-protein interactions without labeling either of the two interacting proteins; this procedure poses minimum effects on the binding properties of the proteins. Our strategy uses a protein/aptamer complex to probe the interactions in a competitive assay where the binding of an aptamer to its target protein is altered by a second protein that interacts with the target protein. Two signal transduction strategies, fluorescence resonance energy transfer (FRET) and fluorescence anisotropy, have been designed to study the interactions of human alpha-thrombin with different proteins by using two aptamers specific for two binding sites on alpha-thrombin. Our method has been shown to be simple and effective, does not require labeling of proteins, makes use of easily obtainable aptamers, provides detailed protein-protein interaction information and has excellent sensitivity for protein detection and protein protein interaction studies. The FRET and the fluorescent anisotropy approaches complement each other in providing insight into the kinetics, mechanisms, binding sites and binding dynamics of the interacting proteins. PMID- 15892185 TI - On the regio- and stereoselective synthesis of aminocyclitols from cyclitol epoxides: the effect of Li as a chelating agent. AB - Experimental and theoretical studies on the influence of Li ions on the regio- and the stereoselectivity of the reaction of cyclitol epoxides with nitrogen nucleophiles have been carried out. Model studies with NaN3 as a nucleophile in the absence of Li ions predict a mixture of C1 and C2 regioadducts. The inclusion of two Li ions as a chelating agent favours the operation of a low populated "all axial" conformation leading ultimately to the C1 adducts. In all cases, the results can be rationalised by geometric and energetic considerations of the corresponding transition states. Predictions of the theoretical calculations are in good agreement with the experimental results using primary and secondary amines as nucleophiles, and thus confirm the validity of this study. PMID- 15892186 TI - Substituent effect to prevent autoxidation and improve spectral stability in blue light-emitting polyfluorenes. AB - A group of fluorene-based polymers, PF-1SOR and PF-2SOR, were synthesized and characterized as blue light-emitting materials. PF-1SOR and PF-2SOR displayed nematic liquid crystalline mesophase in films cast from solution. Compared with conventional polyfluorene, PF-1SOR and PF-2SOR display blue-shifted UV absorption and structureless blue fluorescence. The photoluminescence spectra of PF-1SOR and PF-2SOR were found insensitive against thermal treatment in air up to 200 degrees C and the blue electroluminescence in their light-emitting devices was independent of the driving voltage. Compared to the conventional polyfluorenes, the improved spectral stability of these polymers is attributed to the anti oxidization effect of (3,5-di(tert-butyl)phenoxy)sulfonyl side groups attached to the backbone. PMID- 15892187 TI - Contamination of anaesthetic gases with nitric oxide and its influence on oxygenation. PMID- 15892188 TI - Morphological and biochemical perturbations in rat erythrocytes following in vitro exposure to Fenvalerate and its metabolite. AB - Erythrocytes are a convenient model to understand the membrane oxidative damage induced by various xenobiotic-prooxidants. In this investigation, we have examined the potency of Fenvalerate (FEN) and its metabolite, p-chlorophenyl isovaleric acid (p-CPIA) to induce oxidative stress response in rat erythrocytes in vitro in terms of lipid peroxidation and effects on selected antioxidant enzymes. Susceptibility of erythrocytes to FEN exposure was further investigated in terms of morphological alterations by scanning electron microscopy and protein damage by gel electrophoresis of erythrocyte ghosts. Following in vitro exposure, FEN caused a significant induction of oxidative damage in erythrocytes at concentrations beyond 0.1 mM as evidenced by increased thiobarbituric acid reactive substances (TBARS) levels. The response was both concentration and time dependent. At higher concentrations, significant decreases in the activities of vital antioxidant enzymes viz., catalase, superoxide dismutase, glutathione transferase and glutathione reductase were also discernible clearly suggesting the potency of both, parent compound and its metabolite to induce oxidative stress in erythrocytes. Scanning electron micrographs of erythrocytes following FEN exposure at higher concentrations revealed various degrees of distortion in shape and ruptured membranes. Furthermore, gel electrophoresis studies revealed consistent and significant aggregation of only band 3 protein in erythrocyte membranes exposed to either FEN or p-CPIA at higher concentrations. These in vitro findings show that FEN and its metabolite have the propensity to cause significant oxidative damage in rat erythrocytes, which is associated with marked damage to membrane proteins. These data suggest that both structural and functional perturbations may ensue in erythrocytes following exposure to FEN at higher concentrations under in vivo situations. PMID- 15892189 TI - A green thumb and a broad back: a tribute to the late Dr. Yasuo Inoue (1934 2005). PMID- 15892190 TI - The science of myelin. Proceedings of a symposium of the Society for Experimental Neuropathology under the auspices of the American Neurological Association. October 3, 2004. Toronto, Ontario, Canada. PMID- 15892191 TI - Heart Rhythm 2005. Abstracts of the 26th Annual Meeting of the Heart Rhythm Society. May 4-7, 2005, New Orleans, Louisiana, USA. PMID- 15892192 TI - [Critical observation on the term "complex post-traumatic stress disorder"]. PMID- 15892193 TI - Implantable cardioverter-defibrillators. PMID- 15892194 TI - Implantable cardioverter-defibrillators. PMID- 15892195 TI - Implantable cardioverter-defibrillators. PMID- 15892196 TI - APC is a component of an organizing template for cortical microtubule networks. AB - A microtubule network on the basal cortex of polarized epithelial cells consists of non-centrosomal microtubules of mixed polarity. Here, we investigate the proteins that are involved in organizing this network, and we show that end binding protein 1 (EB1), adenomatous polyposis coli protein (APC) and p150Glued - although considered to be microtubule plus-end-binding proteins - are localized along the entire length of microtubules within the network, and at T-junctions between microtubules. The network shows microtubule behaviours that arise from physical interactions between microtubules, including microtubule plus-end stabilization on the sides of other microtubules, and sliding of microtubule ends along other microtubules. APC also localizes to the basal cortex. Microtubules grew over and paused at APC puncta; an in vitro reconstituted microtubule network overlaid APC puncta; and microtubule network reconstitution was inhibited by function-blocking APC antibodies. Thus, APC is a component of a cortical template that guides microtubule network formation. PMID- 15892197 TI - Angiotensin II type 1-receptor activating antibodies in renal-allograft rejection. PMID- 15892198 TI - Bites of the brown recluse spider. PMID- 15892199 TI - Nevirapine misinformation: will it kill? AB - Three Associated Press (AP) reports in mid-December 2004 created widespread doubts about nevirapine (NVP), the critically important drug that can prevent HIV in many of the 1,800 babies infected every day by their mothers in childbirth. The media allegations that circulated around the world grew out of a bitter personal and personnel dispute between two employees at the US National Institutes of Health (NIH) about the conduct of the HIVNET 012 trial in Uganda from 1997 to 1999. No new information was released about NVP; indeed, following publication of the AP reports, physicians know that NVP still has the same risks and benefits as before. But many experts fear that the emotions stirred by the misinformation disseminated worldwide will result in many HIV-positive mothers receiving no treatment and unnecessarily infecting their children with HIV. This article offers background that has been missing in many news reports. PMID- 15892200 TI - Workshop convened on HIV/hepatitis coinfection. PMID- 15892201 TI - Out of the headlines. PMID- 15892202 TI - 44th ICAAC highlights. PMID- 15892203 TI - Aggressive treatment for unstable CAD improves quality of life. Early intervention relieves angina better than more conventional medical treatments alone. PMID- 15892204 TI - Avoid fried fish, avoid a stroke. Eat your omega-3 fatty acid-enriched fish baked or broiled--and reduce your chance of stroke. PMID- 15892205 TI - The benefits of strength training--at any age. Resistance is not just for athletes and body builders; it's for everyone, regardless of age--including you. PMID- 15892206 TI - I've been using sunscreen for years, but I have noticed some spots and blemishes on my skin. How can I tell what's potentially cancerous and what isn't? PMID- 15892207 TI - States wrangle with corrections budgets: criminal justice budgets challenge states as costs and inmate populations increase. PMID- 15892208 TI - The meth menace: battling the fast-paced spread of methamphetamine may mean attacking it from several fronts. PMID- 15892209 TI - Profile. Mark A. Malangoni, M.D. Interview by Vicki Glaser. PMID- 15892210 TI - Profile. Donald E. Fry. Interview by Vicki Glaser. PMID- 15892211 TI - From crystal structure to clinic: highlights of the Tenth International Parvovirus Workshop. PMID- 15892212 TI - New drugs and dosage forms. PMID- 15892214 TI - You need to know: living wills. PMID- 15892213 TI - Preventing the accumulation of surplus controlled substances at long term care facilities. Final rule. AB - DEA is amending its regulations to allow, where State laws permit, for retail pharmacy installation of automated dispensing systems at long term care facilities. Automated dispensing systems would allow dispensing of single dosage units and mitigate the problem of excess stocks and disposal. PMID- 15892215 TI - Rating the diets from Atkins(TM) to Dr. Sears Zone. PMID- 15892216 TI - Seafood allergies: common, sudden, deadly. PMID- 15892217 TI - Current awareness in prenatal diagnosis. PMID- 15892218 TI - Eight essential traits of leadership. PMID- 15892219 TI - Dental photography systems: required features for equipment selection. AB - Photographic images are becoming a standard of care for visual communication and medicolegal documentation in contemporary dentistry. For visual images to be effective in diagnosis and treatment planning they must capture useful visual information. Repeatable and predictable image alignment, magnification, and exposure are required for predictable macrophotography in dental practice. This article discusses basic photographic principles that influence specific dental imaging needs and the camera system features required to achieve repeatable diagnostic results. PMID- 15892220 TI - New materials and techniques for endodontic success. PMID- 15892221 TI - Evaluation of a dual peak third generation LED curing light. AB - This study compared 3 light-emitting diode curing lights (UltraLume 5, FreeLight 2, LEDemetron I) with a quartz-tungsten-halogen light (Optilux 401) to determine which was the better at photopolymerizing 5 resin composites. The composites were 2 mm thick and were irradiated for the manufacturers' recommended curing times at distances of 2 mm and 8 mm from the light guide. The Knoop hardness at each of 22 points over a 10-mm diameter footprint at the top and bottom of the composites was used to compare the lights. The 4 curing lights and irradiation distances did not have the same effect on all the composites (P < .001). It was concluded that overall the UltraLume 5 dual peak third generation LED curing light was able to polymerize these 5 resin composites as well as or better than the other curing lights. PMID- 15892222 TI - Technology in the endodontic office. PMID- 15892223 TI - New perspectives for diagnosing equine parasitic disease. PMID- 15892224 TI - Cooking the books or simply getting the best out of the data? Assessing the nature of the relationship between variables. PMID- 15892225 TI - Risk factors for fatal lateral condylar fracture of the third metacarpus/metatarsus in UK racing. AB - REASONS FOR PERFORMING STUDY: Lateral condylar fractures of the third metacarpus/metatarsus are the most common cause of equine fatality on UK racecourses. Identification of risk factors for such injuries and the subsequent implementation of intervention strategies could significantly reduce the total number of racecourse fatalities. OBJECTIVES: To identify horse-, race- and course level risk factors for fatal lateral condylar fracture in Thoroughbreds on UK racecourses. METHODS: Case-control study designs were used. Case horses were defined as those that were subjected to euthanasia having sustained an irreparable lateral condylar fracture while racing at any of the 59 UK racecourses. Case races were defined as those in which one or more horses sustained a fatal lateral condylar fracture. Three controls for each case horse were selected at random from the race in which the case was running. Three controls for each case race were selected at random from all races of the same type held in the same year. Ninety-eight cases were included in the study. Conditional logistic regression was used to identify the relationship between a number of independent variables and the likelihood of fracture. RESULTS: Horses doing no gallop work during training and those in their first year of racing were at significantly increased risk of fracture on the racecourse. Case horses were also more likely to have started racing as 3- or 4-year-olds. Fractures were found to be more likely in longer races with a larger number of runners, races in which professional jockeys were not permitted to ride and races in which the going was described as firm or hard. CONCLUSIONS AND POTENTIAL RELEVANCE: Modifications to training schedules, specifically within the first year of racing, may have a large impact on the risk of fatal lateral condylar fracture on the racecourse. Horses should do some gallop work in training and our results suggest that the minimum distance galloped should be between 201 m (1 furlong) and 1609 m (8 furlongs) per week. The association with age at first race requires further investigation for flat and National Hunt racing separately. A reduction in the number of races taking place on very firm going could have an impact on the number of lateral condylar fractures. PMID- 15892226 TI - Study of cartilage and bone layers of the bearing surface of the equine metacarpophalangeal joint relative to different timescales of maturation. AB - REASONS FOR PERFORMING STUDY: A detailed and comprehensive insight into the normal maturation process of the different tissues that make up functional units of the locomotor system such as joints is necessary to understand the influence of early training on musculoskeletal tissues. OBJECTIVES: To study simultaneously the maturation process in the entire composite structure that makes up the bearing surface of a joint (cartilage, subchondral and trabecular bone) in terms of biochemical changes in the tissues of juvenile horses at 2 differently loaded sites of the metacarpophalangeal joint, compared to a group of mature horses. HYPOTHESIS: In all the structures described above developmental changes may follow a different timescale. METHODS: Age-related changes in biochemical characteristics of the collagen part of the extracellular matrix (hydroxylysine, hydroxyproline, hydroxypyridinum crosslinks) of articular cartilage and of the underlying subchondral and trabecular bone were determined in a group of juvenile horses (n = 13) (Group 1, age 6 months-4 years) and compared to a group of mature horses (n = 30) (Group 2, >4 years). In both bony layers, bone mineral density, ash content and levels of individual minerals were determined. RESULTS: In cartilage, subchondral bone and trabecular bone, virtually all collagen parameters in juvenile horses were already at a similar (stable) level as in mature horses. In both bony layers, bone mineral density, ash- and calcium content were also stable in the mature horses, but continued to increase in the juvenile group. For magnesium there was a decrease in the juvenile animals, followed by a steady state in the mature horses. CONCLUSIONS: In horses age 6 months-4 years, the collagen network of all 3 layers within the joint has already attained a mature biochemical composition, but the mineral composition of both subchondral and trabecular bone continues to develop until approximately age 4 years. POTENTIAL RELEVANCE: The disparity in maturation of the various extracellular matrix components of a joint can be assumed to have consequences for the capacity to sustain load and should hence be taken into account when training or racing young animals. PMID- 15892227 TI - Bacterial isolates and antimicrobial susceptibilities in equine bacterial ulcerative keratitis (1993--2004). AB - REASONS FOR PERFORMING STUDY: Bacterial ulcerative keratitis is a common and often vision-threatening problem in horses. Emerging bacterial resistance to commonly used topical antibiotics has been demonstrated. Previous antibiotic use may alter the antimicrobial susceptibility of bacterial isolates. OBJECTIVES: To document aerobic bacterial isolates and associated bacterial susceptibilities from horses with ulcerative keratitis treated at the University of Tennessee between January 1993 and May 2004 and determine whether prior antibiotic therapy affected antimicrobial susceptibility of the isolates. METHODS: Medical records from horses with ulcerative keratitis and positive aerobic bacterial cultures and antimicrobial susceptibility were evaluated. Clinical history regarding antibiotic therapy prior to culture was documented. RESULTS: Fifty-one aerobic bacterial isolates from 43 horses were identified. Streptococcus equi subspecies zooepidemicus was the most commonly isolated organism, accounting for 33.3% of all isolates, followed by Pseudomonas aeruginosa (11.8%), Staphylococcus spp. (11.8%) and Gram-negative nonfermenting rods (7.8 %). No resistance was noted amongst S. equi ssp. zooepidemicus to cephalothin, chloramphenicol or ciprofloxacin. Only 64 % of S. equi ssp. zooepidemicus isolates were sensitive to bacitracin. No resistance was noted among P. aeruginosa to gentamicin, tobramycin or ciprofloxacin. Antibiotic therapy with neomycin-polymixin B-bacitracin prior to presentation and culture was documented in 11/17 horses in which S. equi ssp. zooepidemicus was isolated and in 4/6 horses in which P. aeruginosa was isolated. Three horses received topical corticosteroids prior to culture, of which 2 had polymicrobial infections. CONCLUSIONS: S. equi ssp. zooepidemicus and P. aeruginosa were the most frequently isolated bacterial organisms in equine ulcerative keratitis. No significant trends in aminoglycoside or fluoroquinolone resistance were noted among these organisms. However, the resistance of S. equi ssp. zooepidemicus to bacitracin with common use of this antibiotic suggests that previous antibiotic therapy probably affects antimicrobial resistance. POTENTIAL RELEVANCE: Therapy prior to culture may play an important role in antimicrobial susceptibility of corneal bacterial isolates. Corticosteroid use may increase the risk of polymicrobial infections of corneal ulcers, leading to a worse prognosis. Although significant fluoroquinolone resistance has not been documented in the veterinary literature, these antimicrobials should be reserved for known infected corneal ulcers and not used for prophylaxis. Empirical antibiotic therapy should not only be guided by clinical signs, but also take into consideration previous antimicrobial and corticosteroid therapy. PMID- 15892228 TI - Study of serum amyloid A concentrations as a means of achieving early diagnosis of Rhodococcus equi pneumonia. AB - REASONS FOR PERFORMING STUDY: Prognosis of Rhodococcus equi pneumonia can be challenging because the course of the disease is often insidious and overt clinical signs are subtle. Early diagnosis is considered desirable because it may offer the chance of more successful implementation of treatment and, thereby, improved outcome. Serological tests have previously failed to be accurate for early detection or diagnosis. Measurement of serum amyloid A (SAA) prior to and at the time of clinical signs was therefore chosen in order to assess its potential clinical use. OBJECTIVE: To determine whether SAA concentrations differentiate foals affected with R. equi pneumonia from unaffected foals, either prior to the onset of disease or at the time of onset of clinical signs. HYPOTHESIS: SAA concentrations are significantly higher among foals that develop R. equi pneumonia than in foals from the same environment that remain clinically unaffected. METHODS: Serum samples were obtained from 212 foals 7-14 days and 196 foals 21-28 days post partum, and from affected foals and age-matched controls at the time of onset of signs of pneumonia. SAA concentration was determined for each sample. RESULTS: There were no significant differences between SAA concentrations of foals with R. equi and clinically unaffected foals during the 2 periods of examination or at the time of onset of clinical signs of R. equi pneumonia. CONCLUSIONS: Concentrations of SAA are variable among foals with R. equi pneumonia and cannot be used reliably either as an ancillary diagnostic tool or to screen for early detection of disease during the first month post partum. POTENTIAL RELEVANCE: Bimonthly monitoring concentration of SAA is not useful as a screening test for early detection of R. equi pneumonia and does not facilitate diagnosis of this disease when used according to the protocol of this study. PMID- 15892229 TI - Prevalence of Taylorella equigenitalis infection in stallions in Slovenia: bacteriology compared with PCR examination. AB - REASONS FOR PERFORMING STUDY: The prevalence of Taylorella equigenitalis infection in Slovenia is unknown and methods used to refine identification in these stallions are required. HYPOTHESIS: In diagnosis of T. equigenitalis, polymerase chain reaction (PCR) would have advantages over culture methods, especially in cases where small numbers of causal agent or intensive contamination of genital swabs are involved. METHODS: Culture method and PCR were used to examine a total of 980 genital swabs from the urethra and fossa urethralis of 245 stallions for the presence of the contagious equine metritis organism. RESULTS: Among 245 examined stallions, 225 (91.8%) were negative to T. equigenitalis by both methods. From the swabs of 17 stallions (6.9%) T. equigenitalis was isolated at first and/or second sampling. Swabs of 3 (13%) stallions were PCR positive but the isolation of T. equigenitalis failed. The rate of T. equigenitalis detection was higher with PCR than with the classic bacteriological examination. CONCLUSIONS AND POTENTIAL RELEVANCE: PCR protocol used in this study provided a specific, sensitive, and simple tool for rapid detection of T. equigenitalis. PCR is especially valuable in cases of intensive bacterial and fungal contamination of swabs where the isolation of T. equigenitalis usually fails. PMID- 15892230 TI - Demonstration of regional differences in equine ventricular myocardial velocity in normal 2-year-old Thoroughbreds with Doppler tissue imaging. AB - REASONS FOR PERFORMING STUDY: Doppler tissue imaging (DTI) is a novel noninvasive method by which myocardial velocity can be assessed directly and it allows regional, rather than global, cardiac function to be evaluated. HYPOTHESIS: That regional differences in myocardial velocities exist within the equine ventricle. OBJECTIVES: To develop a repeatable examination technique for DTI in horses, describe DTI findings in various regions of the normal equine ventricle, compare colour (CDTI) and spectral (SDTI) techniques of DTI, and document regional differences in myocardial velocity. METHODS: Five regions of the ventricles (right ventricular wall, interventricular septum and left, right and caudal regions of the left ventricle) were evaluated using SDTI and CDTI in 20 clinically normal Thoroughbreds age 2 years. Individual repeatability of the method was determined by examination of one 6-year-old Thoroughbred on 6 occasions. RESULTS: Three major movements were observed in the ventricular walls in systole, early diastole and late diastole. The interventricular septum had a complex pattern of movement. The left region of the left ventricle and interventricular septum had the most rapid movement. The individual repeatability of CDTI was poor, while in systole and early diastole, but not late diastole, SDTI produced repeatable estimates of maximal myocardial velocity. The different velocity estimates obtained with SDTI and CDTI are not interchangeable. Regional differences in the peak mean and maximal myocardial velocities were found in systole and early diastole (P<0.05), but were not identified in late diastole. CONCLUSIONS: The SDTI modality appears to produce the most repeatable data. There are regional differences in myocardial velocity within the equine ventricles for systole and early diastole. POTENTIAL RELEVANCE: DTI shows potential as a tool for studying regional myocardial movement both in clinical cases suspected of having myocardial dysfunction and in a research setting. In particular, SDTI offers potential as a direct and noninvasive means to study early diastolic function of the equine ventricles. PMID- 15892231 TI - Effects of glucosamine hydrochloride and chondroitin sulphate, alone and in combination, on normal and interleukin-1 conditioned equine articular cartilage explant metabolism. AB - REASONS FOR PERFORMING STUDY: Clinical trials in human and veterinary literature have documented the benefits of oral nutraceutical joint supplements containing glucosamine (GU) and chondroitin sulphate (CS) to treat mild to moderate osteoarthritis, but the effects of these components have not yet been conclusively determined. OBJECTIVES: To assess varying dosages of GU and CS on normal and interleukin-1alpha (IL-1) conditioned equine cartilage explants and rationalise the use of these products. HYPOTHESIS: Treatment would not be detrimental to cartilage metabolism and higher dosages and the combination of GU and CS would be more beneficial than lower dosages and. GU or CS alone. METHODS: Articular cartilage explants collected from the femoral trochlea and condyles were cultured in normal and IL-1 conditioned media. Treatment groups included 0, 12.5, 25,125 and 250 microg/ml concentrations of GU alone, CS alone, or GU+CS in combination. Glycosaminoglycan (GAG) synthesis and total GAG content in the explants and media were analysed. RESULTS: There were no detrimental effects of GU, CS or GU+CS on cartilage metabolism. High dosages of GU+CS reduced total GAG release into the media (degradation). CONCLUSIONS: Our results suggests that GU+CS may prevent cartilage GAG degradation. POTENTIAL RELEVANCE: The combination of GU and CS may be more effective in preventing or treating osteoarthritis in horses than either product alone. PMID- 15892232 TI - Electromyographic activity of the stylopharyngeus muscle in exercising horses. AB - REASONS FOR PERFORMING STUDY: There is a need to understand the process which leads to failure of recruitment of the stylopharyngeus muscle in clinical cases of nasopharygeal collapse. We therefore studied the timing and intensity of stylopharyngeus muscle activity during exercise in horses. OBJECTIVE: To measure the electromyographic (EMG) activity of the stylopharyngeus muscle in exercising horses and correlate it with the breathing pattern. METHODS: Five horses were equipped with a bipolar fine wire electrode placed on the stylopharyngeus muscle and a pharyngeal catheter. The horses exercised on a treadmill at speeds corresponding to 50 (HRmax50), 75 and 100% of maximum heart rate, and EMG activity of the stylopharyngeus muscle and upper airway pressures were recorded. The EMG activity of the stylopharyngeus muscle was then correlated to the breathing pattern and the activity quantified and reported as a percentage of the baseline activity measured at HRmax50. RESULTS: There was ongoing activity of the stylopharyngeus muscle throughout the breathing cycle; however, activity increased towards the end of expiration and peaked early during inspiration. Tonic activity was present during expiration. Peak, mean electrical and tonic EMG activity increased significantly (P<0.05) with exercise intensity. CONCLUSION: The stylopharyngeus muscle has inspiratory-related activity and tonic activity that increases with speed. POTENTIAL RELEVANCE: The stylopharyngeus muscle is one of a group of upper airway muscles that function to support and maintain the patency of the nasopharynx during inspiration. Failure of recruitment of the stylopharyngeus muscle during exercise is a potential explanation for clinical cases of dorsal pharyngeal collapse, but further work investigating the activity of the stylopharyngeus muscle in horses affected by this disease is needed. PMID- 15892233 TI - Inflammatory airway disease, nasal discharge and respiratory infections in young British racehorses. AB - REASONS FOR PERFORMING STUDY: Respiratory disease is important in young Thoroughbred racehorses, but the variation in the rates of occurrence between different ages and training groups has not been characterised. OBJECTIVES: To determine the rates of respiratory disease, particularly inflammatory airway disease (IAD), as well as evidence of infection, and their variation between age and group. METHODS: Horses were examined monthly in 7 British flat training yards over a 3 year period. IAD was defined as increased mucus in the trachea with increased proportions of neutrophils in tracheal wash samples. Frequencies of disease outcomes were estimated from the data. RESULTS: The prevalence of IAD was 13.8% and the incidence was 8.9 cases/100 horses/month. Rates varied with training and age groups, decreasing in older animals. The prevalence of nasal discharge (ND) was 4.1%. Rates of bacterial isolation were more common than viral infections. The incidence and prevalence of several bacterial species decreased with age. CONCLUSIONS: IAD and ND were common in young racehorses, varying significantly between training groups and decreasing with age, consistent with infection playing a role in aetiology. POTENTIAL RELEVANCE: The high prevalence of IAD in 2-year-old horses in Britain suggests that routine endoscopic examination may be helpful in providing early diagnosis and appropriate therapy. The transmission of bacteria and viruses within and between groups of young animals and the role of infection, stable environment and factors inherent to each horse, including their genetic make-up, in the multifactorial aetiology of the disease all merit further study. PMID- 15892234 TI - Antioxidant and inflammatory responses of healthy horses and horses affected by recurrent airway obstruction to inhaled ozone. AB - REASONS FOR PERFORMING STUDY: Inhaled ozone can induce oxidative injury and airway inflammation. Horses affected by recurrent airway obstruction (RAO) have a decreased pulmonary antioxidant capacity, which may render them more susceptible to oxidative challenge. It is currently unknown whether RAO-affected horses are more susceptible to oxidative stress than those unaffected by RAO. OBJECTIVES: To determine whether ozone exposure induces greater oxidative stress and airway inflammation in RAO-affected horses in remission than in healthy horses. METHODS: Seven healthy control horses and 7 RAO-affected horses were exposed to 0.8 ppm ozone for 2 h at rest. RESULTS: At baseline, bronchoalveolar lavage fluid (BALF) ascorbic acid concentrations were lower in RAO-affected horses than healthy controls. Ozone appeared to preferentially oxidise glutathione rather than ascorbic acid 6 h after exposure. Individual healthy and RAO-affected horses demonstrated oxidation of BALF glutathione after ozone exposure. Overall, RAO affected horses did not demonstrate increased oxidative stress following ozone exposure, compared with healthy horses. Ozone did not induce significant airway inflammation in either group. CONCLUSIONS: RAO-affected horses in remission are not more sensitive to ozone despite a decreased pulmonary antioxidant capacity. Sensitivity to ozone appears to be independent of initial pulmonary antioxidant status. POTENTIAL RELEVANCE: Horses with high susceptibility to oxidative stress may benefit from antioxidant supplementation. PMID- 15892235 TI - Influence of repeated arthrocentesis and exercise on synovial fluid concentrations of nitric oxide, prostaglandin E2 and glycosaminoglycans in healthy equine joints. AB - REASONS FOR PERFORMING STUDY: The importance of osteoarthritis (OA) in the horse and the difficulty in its early diagnosis have led to a search for potential biomarkers of joint disease. If the levels of such markers are to be interpreted accurately, clinicians and researchers need to know whether they are influenced by environmental factors and/or interventions such as exercise and repeated arthrocentesis. OBJECTIVE: To investigate the influence of repeated arthrocentesis and exercise on nitric oxide (NO), prostaglandin E2 (PGE2) and glycosaminoglycan (GAG) concentrations in synovial fluid (SF) from normal equine joints. METHODS: SF was collected from the left metacarpophalangeal (MCP), radiocarpal and tarsocrural joints of 16 horses. Half of the horses were exercised and arthrocentesis was repeated 14, 14.5, 17 and 24 days after the start of the exercise programme, in both exercised and control horses. Nitric oxide was determined in SF from the MCP joint only and PGE2 and GAG concentrations were determined in SF from all joints. RESULTS: Repeated arthrocentesis caused an increase in NO concentration in the MCP joint on Day 145, in PGE2 concentrations in the radiocarpal and tarsocrural joints on Day 145 and the release of GAGs into SF of the MCP and radiocarpal joints on Day 17. Exercise resulted in an increase in PGE2 levels in all joints but did not influence the other parameters measured. POTENTIAL RELEVANCE: Repeated arthrocentesis is a potential confounding factor for the use of synovial NO, PGE2 and GAG concentrations as markers of joint disease. Based on this study, such a confounding effect can be avoided if one week or more separates arthrocentesis procedures. Moderate exercise causes a transient rise in PGE2 in SF. PMID- 15892236 TI - Changes to oxfendazole chiral kinetics and anthelmintic efficacy induced by piperonyl butoxide in horses. AB - REASONS FOR PERFORMING THE STUDY: The study of novel pharmacological strategies to control parasitism in horses is required since many parasite species have developed resistance to anthelmintic drugs. OBJECTIVES: To evaluate the effects of piperonyl butoxide (PB) (a metabolic inhibitor) on the plasma availability and enantiomeric behaviour of oxfendazole (OFZ) given orally to horses, and to compare the clinical efficacy of OFZ given either alone or co-administered with PB in naturally parasitised horses. METHODS: Fifteen naturally parasitised crossbred male ponies were allocated into 3 groups (n = 5) and treated orally as follows: Group I (control) received distilled water as placebo; Group II was dosed with OFZ (10 mg/kg bwt); and Group III was treated with OFZ (10 mg/kg bwt) co-administered with PB (63 mg/kg bwt). Jugular blood samples were obtained over 120 h post treatment. Three weeks after treatments, all experimental horses were subjected to euthanasia. RESULTS: The observed maximum plasma concentration (Cmax) and area under the concentration vs. time curve (AUC) values for OFZ increased 3- and 5-fold, respectively, in the presence of PB. The plasma concentration profiles of fenbendazole (FBZ), a metabolite generated from OFZ, were significantly lower after the treatment with OFZ alone (AUC = 0.8 microg x h/ml) compared to those obtained after the OFZ + PB treatment (AUC = 2.7 microg x h/ml). The enhanced pharmacokinetic profiles correlated with increased anthelmintic efficacy. The combination OFZ + PB showed 100% efficacy against mature nematode parasites. The efficacy against cyathostome L3 larvae increased from 94% (Group II) to 98.7% (Group III). Consistently, the number of L4 larvae recovered from OFZ + PB treated horses (Group III) (n = 146) was significantly lower (P<0.05) than that recovered from Group II (n = 1397). CONCLUSIONS: The use of PB as a metabolic inhibitor may be useful to enhance OFZ activity against mature and migrating larvae of different parasite species in horses. POTENTIAL RELEVANCE: Metabolic inhibitors may be used to enhance the activity of benzimidazole anthelmintics and extend the effective lifespan of benzimidazole drugs in the face of increasing resistance. PMID- 15892237 TI - A new tool for the diagnosis in vivo of habronemosis in horses. PMID- 15892238 TI - Survey of cardiac arrhythmias during submaximal and maximal exercise in Thoroughbred racehorses. PMID- 15892239 TI - Sudden death in racing Thoroughbreds in Victoria, Australia. PMID- 15892240 TI - Congenital abnormalities of the bicipital apparatus in four mature horses. PMID- 15892241 TI - A novel, progressive, sclerosing panencephalitis in a horse. PMID- 15892242 TI - Editorial overview: whither the pharmaceutical industry? PMID- 15892243 TI - Editorial opinion: chemoinformatics - a ten year update. AB - The field of chemoinformatics was initially introduced to the pharmaceutical industry in 1995, with the first publication appearing in 1998. Since the introduction of the term and the practice, there has been much debate on the spelling and meaning of chemoinformatics. This article offers a single definition for chemoinformatics, which is supported by the publications of practitioners in the field. Several excellent papers that highlight the use of chemoinformatics are cited. This article, similarly to the first publication, will attempt to project a vision for this field. PMID- 15892244 TI - Current directions in the evolution of compound libraries. AB - Synthetic compound libraries have been available for over a century, but in the last 15 years they have become increasing important in early-stage drug discovery projects. The qualities that these libraries possess impart a dramatic effect on hit discovery, on the subsequent steps of hit-to-lead progression, and on lead optimization by medicinal chemists. More recently, synthetic libraries have found an important application in chemical genetics. Concurrently, ideas have evolved regarding the application of the different types of synthetic approaches that are variously described as combinatorial chemistry, high-throughput synthesis, or diversity-oriented synthesis. This review will discuss current directions of synthetic chemistry, natural product chemistry and chemoinformatics, which relate to the various types of compound libraries that are in use in drug discovery and chemical genetics. PMID- 15892245 TI - Large-scale integrated databases supporting drug discovery. AB - Over the past 15 years, genomics, combinatorial chemistry and high-throughput automation have transformed the setting for drug discovery, from an information poor to a data-rich environment. The next challenge for informatics scientists is to convert the large amount of disparate data produced into useful, integrated information. Consolidation of the different types of information related to drug discovery requires a good working knowledge of database technology, the existence of accepted data standards for achieving uniformity and a complete understanding of the different data systems that are already available. Chemogenomic databases represent the first example of truly integrated systems that make 'omic' technologies directly relevant to small-molecule drug discovery. Researchers within drug discovery programs now have an opportunity to take advantage of new information domains, through the advance and adoption of integrated chemogenomic databases. PMID- 15892246 TI - Advances in knowledge management for pharmaceutical research and development. AB - There are two assumptions that are taken for granted in the pharmaceutical industry today. Firstly, that we can generate an unprecedented amount of drug related information along the research and development (R&D) pipeline, and secondly, that researchers are more connected to each other than they have ever been, owing to the internet revolution of the past 15 years or so. Both of these aspects of the modern pharmaceutical company have brought many benefits to the business. However, the pharmaceutical industry is currently under fire due to allegations of decreased productivity despite significant investments in R&D, which if left to continue at the present pace, will reach almost US 60 billion dollars by 2006. This article explores the role of knowledge in the industry and reviews recent developments and emerging opportunities in the field of knowledge management (KM) as it applies to pharmaceutical R&D. It is argued that systematic KM will be increasingly necessary to optimize the value of preceding advances in high-throughput approaches to R&D, and to fully realize the anticipated increase in productivity. The application of KM principles and practices to the business can highlight opportunities for balancing the current reliance on blockbuster drugs with a more patient-centric focus on human health, which is now becoming possible. PMID- 15892247 TI - Advances in text analytics for drug discovery. AB - The automated extraction of biological and chemical information has improved over the past year, with advances in access to content, entity extraction of genes, chemicals, kinetic data and relationships, and algorithms for generating and testing hypotheses. As the systems for reading and understanding scientific literature grow more powerful, so must the infrastructure in which to assemble information. Advances in infrastructure systems are discussed in this review. Research efforts have flourished as a result of text analytics competitions that attract participants from various disciplines, from computer science to bioinformatics. PMID- 15892248 TI - Feature selection in quantitative structure-activity relationships. AB - A key component of building quantitative structure-activity relationship (QSAR) models is the selection of an appropriate set of molecular features or descriptors. Feature selection can affect the accuracy, stability and interpretability of a model. There are thousands of molecular descriptors currently available, and the selection of an appropriate descriptor set for a particular model can be a daunting task. While there are no absolute rules for selecting appropriate sets of descriptors, a number of recent publications describe automated methods for identifying optimal feature sets. This review provides an overview of a number of the methods described in these publications. PMID- 15892249 TI - From targets to leads: the importance of advanced data analysis for decision support in drug discovery. AB - Lead discovery is a complex process that is intimately linked to chemistry, but which is also increasingly driven by biological sciences. In an industrial pharmaceutical research environment the process is defined by highly automated technologies for target identification and validation, compound library screening, and compound efficacy assessment. The huge volumes and complex dependencies of data produced by such large-scale experiments have led to a reassessment of data analysis processes, resulting in the development of novel data analysis strategies tailored to drug discovery. In this review, recent progress in data-driven research applications is reported, focusing on the use and processing of transcriptomics, proteomics and high-throughput screening data. The successful application of specialized data analysis procedures in many companies is discussed, which has resulted in significant improvements in decision-making processes for progressing therapeutic targets to promising leads. PMID- 15892250 TI - Biochip platforms as functional genomics tools for drug discovery. AB - Improvements in DNA microarray technology have resulted in the generation of data on a scale that, for the first time, permits detailed scrutiny of the human genome. These data provide the foundation for understanding not only the connections between genes and the purpose of genes in the human genome, but also the molecular basis of genetic defects. These advances have the potential to significantly improve healthcare management by improving disease diagnosis and specifically targeting molecular therapy. Herein, the current state of the technology is reviewed, the commercial platforms used by the biopharmaceutical industry are compared and contrasted, and recent efforts in cross-platform data integration are explored. PMID- 15892251 TI - Maximizing discovery efficiency with a computationally driven fragment approach. AB - A reliable and accurate method for the computational design of novel drug candidates has been a passionate pursuit of the pharmaceutical industry. Such technology would dramatically improve the efficiency of drug discovery by quickly and inexpensively providing potent molecules that can be further prioritized for synthesis based on characteristics such as patentability, specific protein-ligand interactions, ease of chemical synthesis, protein selectivity and pharmacological considerations. Described herein is the progress made at Locus Pharmaceuticals Inc toward achieving this ideal with a fragment-driven, computationally directed approach to small-molecule discovery. Specific lead identification examples from Locus Pharmaceuticals discovery programs demonstrate the efficiency and cost effectiveness realized by such an approach. PMID- 15892252 TI - NMR fragment screening: tackling protein-protein interaction targets. AB - High-throughput screening of libraries containing compounds of 'drug-like' molecular weight has frequently resulted in no or poor drug candidates, especially when screening against demanding drug targets such as protein-protein interactions. Fragment-based lead discovery and optimization has evolved as a promising solution to this problem by combining the universal adaptability of low molecular-weight fragments with immediate structural information on fragment binding modes. This review focuses on nuclear magnetic resonance (NMR) fragment screening techniques, which provide a unique combination of medium-throughput, direct binding site information and broad applicability. The utility and exemplary data of chemical shift-detected NMR fragment screening applied to the challenging protein-protein interaction target PDZ domains are summarized. PMID- 15892253 TI - The value of computer-administered self-report data in central nervous system clinical trials. AB - Over the past decade, significant research has been conducted into the development of computer-delivered interviews and scoring methods to assess disease severity in a number of central nervous system indications. In particular, equivalence has been demonstrated between gold standard assessments (such as the Hamilton depression and Hamilton anxiety rating scales) delivered either by computer or by clinicians, in a number of clinical studies. Such computer-based approaches have a number of advantages including the elimination of inter-rater variance and prevention of inclusion bias when inclusion criteria are based upon baseline ratings. In addition, some studies have demonstrated that computer-administered ratings can be more sensitive to the detection of treatment related changes than clinician ratings. Much of this evidence has led to recent US Food and Drug Administration acceptance of certain computer-administered assessments as primary endpoints in depression clinical trials. PMID- 15892254 TI - No immediate pain relief for the pharmaceutical industry. AB - Over many years, the pharmaceutical industry has developed a highly successful model for making new medicines. Currently, there are an estimated 13,000 prescription drugs on the market. However, the model that has served the sector so well for so long is now breaking down. Lack of productivity in the laboratory, the imminent expiry of patents for numerous best selling products, intense competition and a more demanding marketplace, are all signs of a need for a new approach to drug discovery. Consequently, some pharmaceutical executives may feel that they are managing a previously prestigous team, which now find themselves in the relegation zone, and cannot continue under the same model. This article suggests that radical transformation of the business at every level is the only key to long-term success. PMID- 15892255 TI - Synthesis and biological activity of some unsaturated 6-azauracil acyclonucleosides. AB - A useful route is described for obtaining Z and E unsaturated alkylating agents 3 and 4. Coupling 6-azauracils 5 and 6 with unsaturated alkylating agent followed by the deprotection with H+ resin gave acyclonucleosides 11-14 in good overall yields. Unsaturated acyclonucleosides phosphonates 19 and 20 were prepared using potassium carbonate as base and 4-bromobut-2-enyl diethyl phosphonate 16 as the alkylating agent. The introduction of a propargyl group at the N-3 position of acyclonucleosides 7, 8 17, 18, 19, and 20 was achieved using potassium carbonate in DMF. PMID- 15892256 TI - A one-pot synthesis of 1-alpha- and 1-beta-D-arabinofuranosyl-2-nitroimidazoles: synthons to the markers of tumor hypoxia. AB - 1-alpha- and 1-beta-D-Arabinofuranosyl-2-nitroimidazole (alpha-AZA and beta-AZ A) are synthons for a number of potential markers of tissue hypoxia. A one pot synthesis in which 2-nitroimidazole is coupled with a mixture of alpha- and beta 1-O-acetyl-2,3,5-tri-O-benzoyl-D-arabinofuranose in the presence of stannic chloride, followed by deprotection using ammonia/methanol, is described Previously reported conditions for coupling 2-nitroimidazole to 1-alpha bromoarabinofuranose protected by base-hydrolyzable groups afforded alpha-AZA almost exclusively. PMID- 15892257 TI - Synthesis of 5'-triphosphate mimics (P3Ms) of 3'-azido-3',5'-dideoxythymidine and 3',5'-dideoxy-5'-difluoromethylenethymidine as HIV-1 reverse transcriptase inhibitors. AB - 3'-Azido-3',5-dideoxythymidine 5'-phosphonate and 3',5'-dideoxy-5' difluoromethylenethymidine 5'-phosphonate were prepared by multistep syntheses. The nucleoside 5'-phosphonates were converted to their triphosphates and triphosphate mimics (P3Ms) containing beta,gamma-difluoromethylene, beta,gamma dichloromethylene, or beta,gamma-imodo by condensation with pyrophosphate or pyrophosphate mimics, respectively. Inhibition of HIV-1 reverse transcriptase by the nucleoside P3Ms is briefly discussed. PMID- 15892258 TI - Finding keywords for intergenic and gene regions for human genome. AB - The analysis of functionally related sequences for conserved patterns is important for further research of different functional regions. This paper presents an analysis of genes and intergenic sequences from the point of view of linguistics analysis, where gene and intergenic regions are regarded as two different subjects written in the four-letter alphabet [A, C, G, T] and high frequency simple sequences are taken as keywords. A measurement alpha[l(tau)] was introduced to describe the relative repeat ratio of simple sequences. Cutoff values were found for keywords selection. After eliminating "noise," 87 short sequences were selected as keywords for intergenic regions and 76 for gene regions. PMID- 15892259 TI - A neural network for predicting the stability of DNA/DNA duplexes. AB - A back-propagation neural network method has been developed to predict the stability of DNA/DNA duplexes. Calculated Tm with the present parameters fits the experimental values within reasonable errors (AD = 1.86 K, SEP = 1.99151, R2 = 0.9894 for NN1; AD = 1.59667 K, SEP = 2.03824, R2 = 0.99371 for NN2) and it has the advantage that the determinations of thermodynamic parameters are not needed. PMID- 15892260 TI - A novel versatile phosphoramidite building block for the synthesis of 5'- and 3' hydrazide modified oligonucleotides. AB - We introduce a novel versatile phosphoramidite building block for the modification of oligonucleotides (ONs) with acyl hydrazides on the 5'- or 3' terminus, or both. The reaction of these hydrazide functionalized ONs with 4 methoxyphenylaldehyde is demonstrated for solution derivatization. Hydrazides are considered nowadays as promising reactants, which show enhanced reactivity at neutral and slightly acidic conditions and higher stability of yielding products as compared to the aliphatic amines, which are broadly used for ONs derivatization. Our method to introduce hydrazides into ONs employs a phosphoramidite modifier designed to split, during ammonia or lithium hydroxide treatment, into two hydrazides via beta-elimination of a central bis-2 carbonylethoxysulfone unit. It allows the creation of ONs derivatized with a hydrazide moiety at the 5'-, 3'- and both 5'- and 3'-termini, as well as two different hydrazide containing ONs at the same time, viz. in one sequence on the same solid support In latter case one can, for example, synthesize two hydrazide containing ONs, where one is 5'-modified and second one is 3'-modified. PMID- 15892261 TI - Inhibitory mechanisms of 1-(3-C-ethynyl-beta-D-ribopentofuranosyl)uracil (EUrd) on RNA synthesis. AB - 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)uracil (EUrd) is an antimetabolite that strongly inhibits RNA synthesis and shows a broad antitumor activity in vitro and in vivo. In mouse mammary tumor FM3A cells, EUrd is sequentially phosphorylated to its 5'-triphosphate, EUTP, a major metabolite, and the RNA synthesis is inhibited proportionally to its intracellular accumulation. To study the inhibitory mechanisms of EUrd on RNA synthesis, we have performed the kinetic analysis of EUTP on RNA polymerization using isolated nuclei RNA synthesis was inhibited competitively by EUTP. The inhibition constant, Ki was much lower than the Km value of UTP (Ki value of EUTP, 84 nM; Km value of UTP, 13 microM), indicating that the high affinity of EUTP could contribute to the specific inhibition of RNA synthesis. As a result of RNA synthesis inhibition, EUrd, but not ara-C, induced shrinkage of nucleoli, which are the main sites for RNA synthesis in FM3A cells. Thus, the strong affinity of EUTP to RNA polymerase and specific inhibition of RNA synthesis could contribute to its antitumor effect. EUrd is expected to be a new antitumor drug, possessing a strong inhibitory effect on the synthesis of RNA. PMID- 15892262 TI - A letter regarding a case of eosinophilic meningitis. PMID- 15892263 TI - Hawaii's public mental health system. AB - The following article addresses the nature of and problems with the public mental health system in Hawaii. It includes a brief history of Hawaii's public mental health system, a description and analysis of this system, economic factors affecting mental health, as well as a needs assessment of the elderly, individuals with severe mental illness, children and adolescents, and ethnically diverse individuals. In addition to having the potential to increase suicide rates and unnecessarily prolong personal suffering, problems in the public mental health system such as inadequate services contribute to an increase in social problems including, but not limited to, an increase in crime rates (e.g., domestic violence, child abuse), divorce rates, school failure, and behavioral problems in children. The population in need of mental health services in Hawaii is under served, with this inadequacy of services due to economic limitations and a variety of other factors. PMID- 15892264 TI - An assessment of asthma needs in Hawaii. AB - The Hawaii State Asthma Control Program, as a part of state planning, disseminated and collected an asthma needs questionnaire aimed at answering this question: "In your opinion, what asthma-related issues need to be better addressed in Hawaii?" The top five areas of need identified by asthma stakeholders were (1) education, (2) disease management, (3) prevention, (4) healthcare, and (5) support. PMID- 15892265 TI - Geriatric medicine and community health: the Hartford-Reynolds summer program. PMID- 15892266 TI - Medical school hotline: graduate degrees in clinical research at John A. Burns School of Medicine (JABSOM). PMID- 15892267 TI - Cancer research center hotline: PEA-15 phosphoprotein: a potential cancer drug target. PMID- 15892268 TI - [General contemplation on causality in appraising clinical efficacy of traditional Chinese medicine]. AB - This paper briefly introduced the general principles for causality in scientific researches and clinical studies, addressed the importance of population-based approaches in it, and also reviewed the developing course and presented the elementary requirements of randomized controlled trials (RCTs). The author identified with views that as compared with the research design of other types, RCTs is the most authentic one in the aspect of causality of efficacy and intervention. It is stressed that a causal inference for the efficacy assessment of traditional Chinese medicine (TCM) must meet the nomothetic criteria. The author emphasized that the scientific merits of RCTs are not only for assessing the efficacy of conventional treatments, but also for TCM intervention owing to RCTs yield stronger inferences about therapeutic intervention than other study designs, which are wildly accepted by academic communities. PMID- 15892269 TI - [Reports of randomized controlled trials]. PMID- 15892270 TI - [Clinical study on comprehensive treatment of primary liver cancer mainly with chinese medicinal perfusion/embolization]. AB - OBJECTIVE: To study the clinical efficacy and adverse reaction of comprehensive therapy mainly with Chinese anti-cancer medicinal perfusion/embolization and assisted with Chinese drug-therapy based on TCM Syndrome Differentiation in treating primary liver cancer. METHODS: Forty-one patients with liver cancer were divided into the treated group and the control group. To the treated group turmeric oil microballoon, cinobufotalin, Aidi injection and iodized oil were given via hepatic artery perfusion/embolization, and to the control group chemotherapeutic agents and iodized oil were given for instead. Besides, both groups were given Chinese herbs according to TCM Syndrome Differentiation additionally. RESULTS: The tumor inhibitory rate in the treated group and the control group was 77.78% and 69.57% respectively, with insignificant difference between them. The improvement of fatigue and anorexia in the treated group was better than that in the control group (P < 0. 05). The 6-month, 12-month and 24 month survival rate in the treated group and control group was 61.11% vs 56.62%, 27.78% vs 30.43% and 22.22% vs 26.09%, respectively, the difference between the two groups was insignificant. The occurrence of adverse reactions such as decreasing of white blood cells, platelet and hemoglobin, nausea and vomiting were obviously lower in the treated group than those in the control group (P < 0.05). CONCLUSION: Chinese anti-cancer medicinal perfusion/embolization has affirmative short-term clinical effect in treating primary liver cancer with few adverse reactions, which was tolerable to patients, but its long-term clinical efficacy needs further observation. PMID- 15892271 TI - [Efficacy of shenqi fuzheng injection combined with chemotherapy in treatment of acute leukemia and its effect on T-lymphocyte subsets, serum IFN-gamma, IL-10 and IL-2]. AB - OBJECTIVE: To investigate the clinical efficacy of Shenqi Fuzheng Injection (SFI) combined with chemotherapy in treatment of patients with acute leukemia and its effect on the levels of T-lymphocyte subsets (CD4, CD8, CD4/CD8) and serum interferon-gamma(IFN-gamma), interleukin-10 (IL-10) and IL-2. METHODS: Sixty-five patients with initial treating acute leukemia were randomly divided into 2 groups, the SFI group (n = 32) treated with SFI plus chemotherapy (CT), the control group (n = 33) treated with CT only. The remission rate, changes of peripheral mature neutrophilic granulocyte (PMNG) count, T-lymphocyte subsets, serum IL-10 and IL-2 before and after treatment were determined. RESULTS: The remission rate in the two groups showed no obvious difference (P > 0.05). After CT for the 1st, 2nd and 3rd weeks, the PMNG count decreased in both groups, showing significant difference as compared with that before CT (P < 0.01 or P < 0.05). The PMNG count at the end of the 3rd and 4th week of CT remounted to higher than that at 1st and 2nd week, and the increment in the SFI group was significantly higher than that in the control group (P < 0.05). The levels of CD4, CD4 /CD8, IFN-gamma and IL-2 all increased in the two groups after treatment (P < 0.05, P < 0.01), however, that of IL-10 was significantly decreased (P < 0.01). The difference between the two groups in these criteria after treatment was also significant (P < 0.05). CONCLUSION: SFI can improve and regulate the immune function of the patients with acute leukemia undergoing CT, it could promote bone marrow cells proliferation and enhance the efficacy. PMID- 15892272 TI - [Expression of platelet CD62p gene and leucocyte HSP70 gene in patients with blood stasis syndrome MA]. AB - OBJECTIVE: To investigate microcosmic essentials of blood stasis syndrome (BSS) at the gene level. METHODS: One hundred and sixty patients with hypertension or diabetes mellitus, BSS or non-BSS, were strictly selected in disease-syndrome integrated mode according to diagnosis standard. The expression of CD62p gene and HSP70 gene in them were observed, compared and analyzed with blood biochemical indexes by experimental techniques, and compared to those in healthy subjects as control, for exploring the microcosmic mechanism and evolutive rules of BSS formation at the gene level. RESULTS: It was showed, through determination and analysis of the microcosmic indexes closely associated with BSS, that the expression of CD62p gene and HSP70 gene were abnormal in patients with BSS, they were increased in patients with BBS and significantly higher than those in healthy subjects (P < 0.01). The abnormality and level of them were closely correlated with the types of BSS. CONCLUSION: CD62p gene and HSP70 gene might be closely associated with BSS. PMID- 15892273 TI - [Primary exploration on immune associated genome of patients with Pi-Qi deficiency syndrome]. AB - OBJECTIVE: To investigate the abnormal change of immune function in patients with Pi-Qi deficiency Syndrome, and to explore the genomic mechanism of its genesis by cDNA chip techniques. METHODS: The cross probe was made by extracting and microamplifying the total RNA and mRNA of peripheral white blood cells (WBC) in healthy subjects and patients with chronic gastritis and ulcerative colitis, which were labeled by Cy3 and Cy5 respectively. Then equal quantity of the two labeled probes were mixed and hybridized with cDNA chip, fluorescent signal of the chips were scanned with scanner. Data obtained were analyzed for comparing the difference of the expressive levels of immune associated genome in peripheral WBC in healthy subjects with those in patients. RESULTS: Expressions of CD9, CD164, PF4 and RARB gene in WBC of patients, both gastritis and colitis, were down-regulated while those of IGKC, DEFA1 and GNLY were up-regulated. CONCLUSION: The genesis of Pi-Qi deficiency syndrome has its immune associated genomic basis, and the immune functions are disordered in patients with that syndrome. PMID- 15892274 TI - [Comparative analysis on results of treadmill test in patients of coronary heart disease caused angina pectoris with Qi-Yin deficiency syndrome with or without accompanied phlegm and blood stasis syndrome]. AB - OBJECTIVE: To explore the objective special features and role of various indexes of treadmill exercise test (TET) in patients of coronary heart disease (CHD) caused angina pectoris with Qi-Yin deficiency syndrome (QYD) with or without accompanied phlegm and blood stasis syndrome (PBS), to provide references for preventing and treating CHD in clinical practice. METHODS: One hundred and one patients, whose diagnosis measured to the diagnostic standard and the inclusion criteria of angina pectoris and CHD, were classified according to their TCM syndrome type to two groups, the QYD without PBS group (49 cases) and the QYD with PBS group (52 cases). TET was conducted on all the patients. The relative parameters were measured and compared. RESULTS: As compared with the QYD without PBS group, in the QYD with PBS group, (1) the TET positive rate was higher; (2) total exercise time was lesser; (3) the maximal metabolic equivalent (Max MET) was lower; (4) the average depression of ST segment at the exercise endpoint of test (mV) was higher; (5) the time of ST segment depressed for 0.1mV (min) was longer; (6) the metabolic equivalent during ST-segment depressed by 0.1mV was shorter; and (7) the change of QRS wave time-limit before and immediately after TET was more evident. Moreover, in the testing time more patients revealed angina episode after exercise, and less patients had their heart rate reached the requirement in the QYD with PBS group than those in the QYD without PBS group. Comparison between the two groups in all the above-mentioned indices showed significant difference respectively (P < 0.01 or P < 0.05). CONCLUSION: Patients with CHD caused angina pectoris of QYD with PBS are worse in the tolerance for exercise and severer in pathological change of coronary artery than those in those without PBS, they belong to the severe phase of TCM syndrome. PMID- 15892275 TI - [Effect of shenmai injection on immunologic function in patients with dilated cardiomyopathy]. AB - OBJECTIVE: To investigate the effect of Shenmai Injection (SMI) on immunologic function in patients with dilated cardiomyopathy (DCM). METHODS: Fifty-six patients were divided into two groups, the control group treated with conventional western medicine, and the SMI group treated with conventional western medicine plus SMI. The indices including red blood cell (RBC) C3b receptor rosette (RBC-C3bRR) and immune complex rosette (RBC-ICR), T-lymphocyte subsets (CD3, CD4, CD8, CD4/CD8) were determined before and after treatment. RESULTS: The level of RBC-C3bRR, CD4, CD8 and CD3 in patients with DCM were significantly decreased (P <0 .01, P < 0.05), RBC-ICR and CD4/CD8 were significantly increased than those in the normal control group (P < 0.01); While the level of RBC-C3bRR, CD4, CD8 and CD3 in the SMI group after treatment were significantly higher, and the level of RBC-ICR and CD4/CD8 were significantly lower (P < 0.01, P < 0.05) than those in the control group. CONCLUSION: The RBC immune adherence and cellular immune function are lower in patients with DCM, and SMI has the effect in regulating immune function in patients with DCM. PMID- 15892276 TI - [Effect of erigeron injection on platelet level of CD62p and serum content of TNF alpha and IL-6 in patients with acute cerebral infarction]. AB - OBJECTIVE: To investigate the clinical effect of Erigeron injection (El) on positive expression rate of CD62p in platelet and content of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in serum of patients with acute cerebral infarction (ACI). METHODS: Sixty-eight patients with ACI were randomly divided into the treated group (n = 35) and the control group (n = 33). Conventional treatment were given to both groups, and EI 40 ml/d were given additionally to the treated group, the treatment course for both groups was 15 days. The positive expression of platelet CD62p and the serum TNF-alpha and IL-6 in patients before and after treatment were determined with flow cytometric (FCM) and electrochemical-luminescence (ECL) techniques respectively. RESULTS: The total curative effect in the treated group were significantly higher than that in the control group (P < 0.05). Levels of platelet CD62p and serum TNF-alpha and IL 6 in ACI patients before and after treatment were significant higher than those in the healthy group (P < 0.05), all the three parameters were significantly decreased after treatment, and the lowering in the treated group was more significant than that in the control group (P < 0.05). CONCLUSION: The effect of El on ACI patients may relate to its action in down-regulating the expression of platelet CD62p, alleviating the immune response and inflammatory injury of central nervous system induced by cytokines. PMID- 15892277 TI - [Clinical observation on chronic obstructive pulmonary disease of acute aggravating stage treated with activating blood circulation to remove blood stasis recipe]. AB - OBJECTIVE: To investigate the clinical efficiency of activating blood circulation to remove blood stasis recipe (ARR) in treatment of patients with chronic obstructive pulmonary disease (COPD) of acute aggravating stage. METHODS: Adopting the prospective randomized controlled method, sixty patients with COPD of acute aggravating stage were divided into two groups. Patients in the treated group (n = 32) was subdivided into four subtypes, Phlegm-Heat accumulation in Fei type, Phlegm-Dampness accumulation in Fei type, Fei-Shen Yin deficiency type and Fei-Pi Qi deficiency type. They were treated with ARR with modification according to their Syndrome type. Patients in the control group (n = 28) were treated with conventional western medicine. The therapeutic course of the two groups was 2 weeks. Changes of indexes including hemorrheological indexes (blood viscosity) and pulmonary function (FEV1, FEV1/FVE%) were observed before and after treatment. RESULTS: The total effective rate was 90.6% in the treated group, while it was 67.9% in the control group. The symptom score and pulmonary function in the two groups were significantly different after treatment (P < 0.01, P < 0.05). In addition, the effect of ameliorating whole blood viscosity in the treated group was better than that in the control group, showing significant difference (P < 0.05). CONCLUSION: Both therapies can improve clinical symptom in patients, but it was showed the effect in those treated with conventional western medicine plus ARR was better than in those treated with conventional western medicine alone. PMID- 15892278 TI - [Clinical observation on effect of gufusheng capsule in treating early stage Legg Calve-Perthes disease]. AB - OBJECTIVE: To evaluate the clinical efficacy of Gufusheng capsule (GFSC) in treating early stage Legg-Calve-Perthes disease. METHODS: Adopting randomized single controlled trial, 45 cases with Legg-Calve-Perthes disease in early stage were randomly divided into 2 groups, 23 patients in the control group were treated with arthrectomy hip synovial membrane excision of affected lateral, femoral head decompression and transplantation of muscle-bone flap from ileum, above treatment were also given to the 22 patients in the treated group but combined with orally taking of GFSC. The treatment course for both groups was 6 months. RESULTS: Short-term effect after 6 months' treatment showed that the total effective rate in the treated group and the control group was 90.9% and 78.3% respectively, and the excellent rate was 72.7% and 60.9% respectively. The pain visual analogue scoring, clinical symptom and syndrome scoring markedly decreased before and after treatment, showing significant difference between the two groups (P < 0.05, P < 0.01). Long-term effect after 1-year to 4-year follow up showed that the total effective rate and excellent rate between the treated group and the control group was significantly different ( chi2 = 8. 5976, P < 0.05). CONCLUSION: GFSC has marked effect in alleviating pain and ameliorating function of hip joint, being an effective compound recipe for treatment of early stage Legg-Calve-Perthes disease. Therefore, it is worthy of researching and developing furthermore. PMID- 15892279 TI - [Effect of yigu capsule contained serum on mRNA and protein expression of estrogen receptor in osteoblast rats]. AB - OBJECTIVE: To explore the effect of Yigu capsule contained serum on mRNA and protein expression of osteoblast estrogen receptor (ER), for investigating the mechanism of its preventing and treating osteoporosis by means of regulating estrogen. METHODS: Osteoblasts separated from newborn SD rats were cultured, and divided into 3 groups after being passaged, i.e. the drug-serum treated group, the blank serum group and the DMEM medium control group. The relative amount of ERalpha and ERbeta expression were determined with RT-PCR, the affinity (expressed by equilibrium dissociation constant, KD) and number of ER (RT) were analysed by radioligand assay. RESULTS: The relative amount of ERbeta mRNA expression were increased in the drug-serum group, with significant difference as compared with that in the other two groups (P < 0.05), but no significant difference among the three groups in ERa mRNA expression (P > 0.05). KD in the drug-serum group showed insignificant difference as compared with that in the other two groups (P > 0.05), but RT increased in the drug-serum group and the difference in the three groups was significant (P < 0.01). CONCLUSION: Drug contained serum of Yigu capsule can up-regulate the expression of osteoblast ERbeta mRNA and increase the amount of ERs. Regulating estrogen is possibly one of the mechanisms of Yigu capsule in preventing and treating osteoporosis. PMID- 15892280 TI - [Effect of fuzheng yiliu granule on H22 tumor cell apoptosis, p53 and caspase-3 gene expression in mice]. AB - OBJECTIVE: To explore the inhibitory effect of Fuzheng Yiliu Granule (FYG) on growth and apoptosis of H22 tumor cell, and expressions of p53 and Caspase-3 gene. METHODS: The tumor inhibitory rate of FYG on H22 tumor cell line was observed in vivo, cell apoptosis rate and cell cycle were determined by flow cytometry (FCM), and expressions of wild type p53 and Capase-3 mRNA were determined by RT-PCR. RESULTS: FYG could inhibit the growth of H22 tumor cell at the dose of 12g/kg, 24g/kg, the maximal inhibitory rate up to 51.24% (P < 0.01). By FCM, it was shown that FYG could significantly enhance apoptosis rate of cell line H22, with the peak reached 15.84% (P < 0.01), and cause the tumor cell cycle being blocked at G0/G1 phase, with decrease of cells in S phase. RT-PCR illustrated that FYG could significantly up-regulate the level of p53 and Caspase 3 mRNA expression. CONCLUSION: FYG can significantly inhibit the growth of tumor cell in mice, its anti-tumor mechanisms may relate to the cell apoptosis induction, cell cycle regulation and wild type p53 and capase-3 gene expression enhancing. PMID- 15892281 TI - [Effect of danshensu and ligustrazine on related genes of myocardial hypertrophy induced by angiotensin II]. AB - OBJECTIVE: To study the effect of Danshensu (DSS) and Ligustrazine (TMZ), the extracts of Chinese herbs for promoting blood circulation, on angiotensin II (Ang II) induced myocardial hypertrophy and its related genes, and to explore the mechanisms of inhibitory effect. METHODS: Adopting one-step method, the total RNA of myocardial cells was extracted by TRIzol reagent. Then the expression of ANP and beta-actin mRNA, as symbol of myocardial cells, were detected by RT-PCR. RESULTS: Molecular biological research showed that Ang II could significantly increase the expression of ANP mRNA in myocardial cells (P < 0.01), which could be significantly inhibited by Losartan (P < 0.01), both DSS and TMZ had the inhibitory effect (P < 0.05). Ang II could increase beta-actin mRNA expression in myocardial cells simultaneously, Losartan, DSS and TMZ could also significantly inhibit it (P < 0.05). CONCLUSION: The effective ingredients of Chinese herbs for promoting blood circulation, DSS and TMZ, have the effect of inhibiting the hyper expression of ANP and beta-actin induced by Ang II, and preventing myocardial hypertrophy, therefore, it could be used to prevent and treat cardiomegaly. PMID- 15892282 TI - [Effect of drugs for promoting blood circulation on blood lipids and inflammatory reaction of atherosclerotic plaques in ApoE gene deficiency mice]. AB - OBJECTIVE: To observe the effect of six common Chinese medicinal herbs for promoting blood circulation, including Radix Paeoniae rubra (I), Radix Salviae miltiorrhizae (II), Rhizoma Chuanxiong (III), Radix Notoginseng (IV), Semen Persicae (V) and wine steamed Radix et Rhizoma Rhei (VI), on blood lipids and inflammatory reaction of atherosclerotic plaques in ApoE gene deficiency mice. METHODS: Ninety mice, 6 - 8 weeks old, were divided into 8 groups, the model group, the control group (treated with simvastatin) and the six treated groups treated with the above-mentioned 6 Chinese medicinal herbs respectively. All the mice were fed with the diet of western kind for 13 weeks until the mature atherosclerotic plaques formed in them. Then they were treated with respective drugs for another 13 weeks except those in the model group. All the mice were sacrificed at the end of experiment, their blood was collected for lipids determination, heart and aorta were taken out for determining the level of CD68 in root of aorta, as well as the expressions of monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-a (TNF-a) by immunohistochemistry staining. RESULTS: All the 6 Chinese herbs showed regulatory action on blood lipids. The positive expression of CD68 in the model group displayed the highest activity. As compared with the model group, the CD68 positive expressed cells in the control group and the groups treated with Chinese herbs II, III, and IV were lesser (P < 0.05), and the expression of inflammatory factors (MCP-1 and TNF-alpha) in atherosclerotic plaques was significantly lower in the control group and the group treated with Chinese herb VI (P < 0.05). CONCLUSION: Chinese medicinal herbs tested in this study can interfere the maturing progress of atherosclerotic plaques and stabilize the plaques in ApoE deficiency mice, the mechanisms may relate to its actions in regulating lipids metabolism and inhibiting inflammatory reaction. Different Chinese medicinal herbs for activating blood circulation of conventional dosage might show difference in potency and acting links. PMID- 15892283 TI - [Effect of shexiang baoxin pill in alleviating myocardial fibrosis in spontaneous hypertensive rats]. AB - OBJECTIVE: To investigate the effects and mechanisms of Shexiang Baoxin Pill (SBP) on myocardial fibrosis in spontaneous hypertensive rats (SHR). METHODS: SHR of 12 weeks old were divided into the SBP group, the control group (treated with benazepril) and the model control group. The effects on such indexes as systolic blood pressure (SBP), left ventricular mass (LVM), left ventricular mass index (LVMI), content of myocardial collagen (MC) in left ventricle, extracellular matrix fibronectin (FN), laminin (LN), cardiac fibroblast (cFb) and transforming growth factor-beta1 (TGF-beta1) were determined after 12 weeks of treatment. RESULTS: SBP had no marked pressure depressive effect, but had the effect similar to that of benazepril in reducing the level of LVM, LVMI and content of MC (P < 0.05), as well as the content of LN, FN in myocardium, cFb count and TGF-beta1 expression (P < 0.05). CONCLUSION: SBP can prevent and treat myocardial fibrosis, whose action is independent of its hypotensive effect. The mechanism may be associated with such factors as the decrease of MC synthesis in left ventricle and the deposition of extracellular matrix. PMID- 15892284 TI - [Toxicity attenuation and efficacy potentiation effects of sijunzi decoction on bladder carcinoma treated by chemotherapy in mice]. AB - OBJECTIVE: To investigate the toxicity attenuation and efficacy potentiation effects of Sijunzi decoction (SJZD) on bladder carcinoma treated by chemotherapy in mice. METHODS: T739 mice were randomly divided into 8 groups after subcutaneous inoculation of bladder carcinoma cells, the control group (A); two mitomycin C (MMC) group, treated with MMC of routine dosage (B) and low-dosage (C) respectively; three SJZD groups, treated with SJZD of high (D), medium (E) and low-dosage (F) respectively; and two combined treatment groups, treated with SJZD of high-dosage + MMC of routine dosage(G) and SJZD of high-dosage + MMC of low-dosage(H). The medication was begun at 24 hrs after inoculation. The tumor inhibitory rate, activity of peritoneal macrophages after 14 days of treatment and change of peripheral white blood cells after 7 days of treatment were determined and the survival time of mice was observed. RESULTS: The survival time of mice in Group D was significantly higher than that in Group A (P < 0.05), while those in Group E and F showed insignificant difference as compared with those in Group A (P > 0.05). The highest tumor inhibitory rate was shown in Group B, but the survival time in that group showed no significant difference as compared to those in Group A (P > 0.05). The longest survival time (32.7 +/- 1.3 days) was shown in Group H, which was obviously different to that in other groups (P < 0.05). And the leukocyte counts and macrophage activity in Group H were better than those in Group B, C and G (P < 0.05), except that the tumor inhibitory rate was significantly lower than that in Group B, C and G (P < 0.05). CONCLUSION: Combined chemotherapy of SJZD with low dosage MMC has definite effect in inhibiting tumor growth in mice with bladder carcinoma, displaying special effects of toxicity attenuation and efficacy potentiation. PMID- 15892285 TI - [Clinical observation on triple-therapy for terminating ectopic pregnancy]. AB - OBJECTIVE: To select a safe, quick and effective method for terminating ectopic pregnancy (EP) with few adverse effects. METHODS: Patients were divided into 2 groups. The observed group was treated with methotrexate (MTX, 50 mg/m2 for single dose intramuscular injection) plus RU 486 (600 mg taken orally in the morning with empty stomach, followed with fasting for 2 hrs) and Waiyun Zhuyu decoction (WZD, one dose a day for 7-10 days). The control group was only treated with MTX, with the same regimen as used in the observed group. RESULTS: The curative rate, time for blood beta-HCG recovering, lump absorption time, and tube recanalization rate in the observed group were better than those in the control group (P < 0.01). CONCLUSION; The key links for successfully treating EP with conservative drug therpy are to diagnose the disease early and clearly, and to select indicative subject strictly. The scheme of single dose administration of MTX plus RU486 combined with WZD, with its high efficacy and few adverse reaction, may be used as the first choice for referential clinical drugs administration. PMID- 15892286 TI - [Clinical observation on effect of Chinese herbs in adjusting hypoestrogenemia status by clomiphene to promote ovulation]. AB - OBJECTIVE: To explore the effect of Yijing Huoxue Cuyun Decoction (YHCD) in adjusting hypoestrogenemic response induced by clomiphene. METHODS: Infertile patients caused by ovulation disturbance were randomly divided into 2 groups. The 60 patients in the observed group were treated with clomiphine plus YHCD, and the 58 patients in the control group were given clomiphine plus estradiol valerate. RESULTS: By scoring on the cervical relaxation and improvement of cervical mucus, 38 patients (63.3%) in the observed group had Insler score of more than 8 points, while that in the control group was only 25 (43.1%), comparison between the two groups showed significant difference (P < 0.05). The endometrium thickening in the observed group was 0.98 +/- 0.19 cm, significantly different to that in the control group (0.85 +/- 0.21 cm, P < 0.01). Twenty-five patients in the observed group (41.7%), and fourteen patients in the control group (24.1%), respectively got pregnancy, the pregnant rate in the former was obviously higher than that in the latter (P < 0.05). CONCLUSION: YHCD can ameliorate hypoestrinemia induced by clomiphene and increase the pregnant rate in patients. PMID- 15892287 TI - [Clinical observation on treatment of non-gonococcal cervicitis by integrative medicine]. AB - OBJECTIVE: To explore the integrative medicinal therapy for non-gonococcal cervicitis (NGC) in order to elevate the therapeutic effect for patients treated in vain after long-term application of antibiotics. METHODS: Fifty patients with NGC were treated with Qingyuan decoction combined with antibiotics, and other 46 patients were treated with antibiotics alone for control. RESULTS: The cure rate and significant effective rate was 90.0% (45/50) and 65.2% (30/46) in the treated group and the control group, it was significantly different between the two groups (chi2 = 9.58, P < 0.01). The disappearance rate of symptom was 88.1% and 41.4% in the two group after treated for two weeks respectively, and it was also significantly different (chi2 = 12.42, P < 0.01). CONCLUSIONS: The therapeutic effect of NGC treated by sensitive antibiotics combined with Qingyuan decoction is better than that treated with western medicine only. PMID- 15892288 TI - [Discussion on thoughts and methods for treatment of diabetic nephropathy by TCM according to inflammatory pathogenesis]. AB - Aim of this article was to investigate relationship between inflammatory pathogenesis of diabetic nephropathy and the TCM pathogenetic theory of Shen Collateral impaired by Toxin, and to illustrate the method for removing toxin, activating collateral and protecting Shen can be an effective treatment for inhibiting the inflammatory pathogenesis of diabetic nephropathy. PMID- 15892289 TI - [English translation of Chinese Journal of Traditional Chinese Medicine]. PMID- 15892290 TI - [Progress on painful diabetic peripheral neuropathy treated by integrative medicine]. AB - The article reviewed clinical studies on painful diabetic peripheral neuropathy (PDPN) treated by integrative medicine. PDPN, a common complication of diabetes mellitus, which could severely influence patients' quality of life. The keystone and difficulty of PDPN treatment is to relieve pain. Tricyclic anti-depressants are the firstline agents for neuropathic pain but with obvious adverse reactions. Antiepileptic drugs and capsicin can relieve PDPN with less adverse reactions. In recent years, lots of report of clinical studies on DPN treated by TCM or integrative medicine were issued, but those pertinent to PDPN were seldom. Only the papers with independent statistical analysis on effect of pain relieving were selected to review in this article, and the authors presumed that it is feasible to treat PDPN with integrative medicine. PMID- 15892291 TI - [Minutes of the 1st International Conference on Rheumatism of Integrative Medicine]. PMID- 15892292 TI - Autosomal-recessive Charcot-Marie-Tooth diseases. AB - In certain countries around the Mediterranean basin such as Algeria, which have a high prevalence of consanguineous marriages, autosomal-recessive (AR) inheritance may account for more than 50% of all forms of Charcot-Marie-Tooth (CMT) disease. Like with the dominant forms, it is usual to differentiate the demyelinating forms (CMT 4 corresponding to autosomal-recessive CMT 1 [AR-CMT 1] from the axonal forms [AR-CMT 2]). Genetic analysis of large families with recessive transmission has uncovered novel CMT genotypes (genes: GDAP 1, MTMR 2, MTMR 13, KIAA1985, NDGR1, periaxi, lamin). The clinical and especially the histologic phenotypes often indicate that a specific gene is implicated. We present and discuss microscopic lesions seen on nerve biopsies from patients in a number of consanguineous Algerian families, and we outline the characteristic lesions that would prompt a search for mutations in genes such as MTMR 2, MTMR 13, KIAA1985, periaxin for CMT 4, and lamin for AR-CMT 2. Like with the dominant forms, there are undoubtedly many more mutations of other genes to be discovered. PMID- 15892293 TI - Spread of T lymphocyte immune responses to myelin epitopes with duration of multiple sclerosis. AB - Although the primary cause of multiple sclerosis (MS) is unclear, evidence supports a role for autoimmune attack of myelin by T lymphocytes. However, it has been difficult to relate patterns of autoimmunity to pathogenesis. In mouse models, the case has been made for relapsing and remitting disease driven by epitope spread: an initial lesion leads to presentation of central nervous system antigens, in turn triggering the next wave of autoimmune T cells of different specificity, the response thus broadening. Few studies have been done to determine whether these events could be important over the longer time scale of human disease. We compared T cell responses with a panel of myelin epitopes in clinically isolated syndrome patients with a first attack, patients with MS with a mean disease duration of 0.95 years, and patients with MS having a mean disease duration of 15.9 years. T cells from patients with long-term disease recognize more myelin epitopes than patients with recent-onset disease. The epitope myelin basic protein 131-149, in particular, was more commonly recognized by patients with long-term disease. The data support the notion that the T cell response in MS broadens with time and is thus implicated in the ongoing pathogenic process. However, there was no clear correlation between disease severity and number of epitopes recognized. This may argue against a simple causal role of epitope spread in driving progression, as has been suggested in experimental allergic encephalomyelitis. PMID- 15892294 TI - Decreased neprilysin immunoreactivity in Alzheimer disease, but not in pathological aging. AB - Although evidence suggests that extensive cortical beta-amyloid (Abeta) deposition is essential in Alzheimer disease (AD), it is also detected in nondemented elderly individuals with pathologic aging (PA). Given evidence that neutral endopeptidase (NEP) or neprilysin, a key enzyme for clearance of Abeta, is decreased in AD, the goal of the present study was to determine if NEP was also decreased in PA. We measured NEP immunoreactivity in frontal cortex of 12 AD and six PA cases and compared this with 10 normal (N) elderly individuals. None had any significant other pathology, and they were similar with respect to age, sex, and postmortem delay. In addition, Abeta1-40 and Abeta1-42 were measured by enzyme-linked immunosorbent assay (ELISA), whereas tau, synaptophysin, and alpha synuclein were measured on Western blots. The AD cases had more neuritic plaques, neurofibrillary tangles, higher Braak stage, and more tau immunoreactivity in frontal cortex than both PA and N. In contrast, both PA and AD had more senile plaques and Abeta1-42 than N. NEP immunoreactivity was decreased in AD but not in PA. The decrease was unlikely the result of neuronal or synaptic loss because NEP immunoreactivity in frontotemporal degeneration with comparable degrees of synaptic loss as the AD cases was not different from control subjects. Although NEP enzyme activity was decreased in approximately half the AD cases, on average, it was not decreased compared with N or PA. The results add further evidence that PA is distinct from AD and indicate that decreased Abeta degradation by NEP is unlikely to contribute significantly to amyloid deposition in PA or, in many cases, of AD. PMID- 15892295 TI - IgG neuropathy: an immunoelectron microscopic study. AB - There are many potential mechanisms of peripheral nerve impairment by a monoclonal IgG dysglobulinemia. In this study, using electron microscopy, we observed widening of the myelin lamellae comparable to that commonly described in IgM neuropathies with antimyelin-associated glycoprotein activity. Such features have yet to be described in IgG neuropathies. In addition, we observed deposits of a granular material in the interstitial tissue of the nerve. By immunoelectron microscopy, we demonstrated the presence of the IgG in the lesions of myelin and the endoneurial space. A direct link between monoclonal dysglobulinemia (regardless of type) and polyneuropathy should be confirmed by nerve biopsy, because the result may influence treatment. PMID- 15892296 TI - INI1 protein expression distinguishes atypical teratoid/rhabdoid tumor from choroid plexus carcinoma. AB - Central nervous system atypical teratoid/rhabdoid tumor (AT/RT) and choroid plexus carcinoma (CPC) are rare, highly malignant tumors that predominantly arise in infants and young children. Overlapping clinical, histologic, ultrastructural, or immunophenotypic features may obscure the diagnosis in some cases. AT/RT is characterized by deletions and/or mutations of the INI1 tumor-suppressor gene on chromosome band 22q11.2. We have recently developed an INI1 immunohistochemical staining assay. Negative staining of tumor cells resulting from inactivation of the INI1 gene is a consistent feature of AT/RT. Mutations of INI1 in some CPCs have been reported. The purpose of the present study was to determine if immunohistochemical staining with an INI1 antibody would provide a sensitive means of distinguishing between CPC and AT/RT. We examined 28 tumors with a submitted diagnosis of CPC. Twenty-one CPCs showed retained expression of INI1 and seven tumors showed loss of INI1 expression. Cytogenetic, FISH, and/or INI1 mutation results were also available for 13 tumors. In three of the seven cases, monosomy 22 was the only cytogenetic abnormality, suggestive of AT/RT. However, monosomy 22 was also identified in 3 tumors with complex karyotypes that retained INI1 expression. The 7 tumors that were immunonegative for INI1 had features that were consistent with AT/RT. Immunostaining for INI1 protein is retained in the majority of CPC and is lost in AT/RT. This expression pattern seems to better define the 2 groups of tumors than does light or electron microscopy, routine immunohistochemistry, or cytogenetic analysis alone. PMID- 15892297 TI - The p15(INK4b)/p16(INK4a)/RB1 pathway is frequently deregulated in human pituitary adenomas. AB - Pituitary adenomas are common benign intracranial neoplasms. However, their tumorigenesis is not yet clearly defined. Inactivation of genes involved in the negative cell-cycle regulatory p15(INK4b) - p16(INK4a) -cyclin D/CDK4-RB1 mediated pathway (RB1 pathway) is one of the most common and important mechanisms in the growth advantage of tumor cells. Recently, much attention has been focused on the importance of alternative mechanisms of gene inactivation, particularly promoter hypermethylation in the transcriptional silencing of such tumor suppressor genes. Based on the rare occurrence of inactivation by gene mutations and deletions of the RB1 pathway in pituitary adenomas, we investigated the deregulation of the RB1 pathway in 42 sporadic human pituitary adenomas, especially focusing on the methylation status of this pathway as determined by a methylation-specific polymerase chain reaction assay. Homozygous deletion of the p15(INK4b) or p16(INK4a) gene was detected in one adenoma each. Amplification of the CDK4 gene was not apparent in any of the pituitary adenomas presently examined. Promoter hypermethylation of the p15(INK4b), p16(INK4a), and RB1 genes was detected in 15 (35.7%), 30 (71.4%), and 12 (28.6%) of the adenomas, respectively. Promoter hypermethylation of the p15(INK4b) gene coincided with p16(INK4a) alteration and/or RB1 methylation, whereas p16(INK4a) and RB1 methylations tended to be mutually exclusive (p = 0.019). Thus, the vast majority of the adenomas (38 of 42, 90.5%) displayed alterations of the RB1 pathway. None of the clinicopathologic features, including the proliferation cell index, was significantly correlated with any particular methylation status. Our results suggest that inactivation of the RB1 pathway may play a causal role in pituitary tumorigenesis, with hypermethylation of the p16(INK4a) gene being the most common deregulation, and further provide evidence that RB1 and p16(INK4a) methylations tend to be mutually exclusive but occasionally coincide with p15(INK4b) methylation. PMID- 15892298 TI - Differential expression of sonic hedgehog immunoreactivity during lesion evolution in autoimmune encephalomyelitis. AB - The signaling molecule Sonic hedgehog (Shh) is involved in several processes of central nervous system development. Recent reports indicate that Shh expression plays a role also in certain pathologic conditions in the adult brain, including multiple sclerosis and its animal model. However, the role of Shh signaling in immune-mediated demyelinating disease remains still uncertain. The aim of our study was to investigate the distribution pattern of Shh immunoreactivity (Shh IR) during lesion evolution in myelin-oligodendrocyte-glycoprotein-induced experimental autoimmune encephalomyelitis (MOG-EAE), a model strongly mimicking multiple sclerosis. MOG-EAE was actively induced in DA rats. Histologic evaluation was performed with light and confocal microscopy on paraffin-embedded central nervous system sections from days 20 to 120 after active immunization. Shh-IR was present within the lesions of MOG-EAE during all stages of lesion evolution. The highest staining intensity for Shh was found in remyelinating lesions. In actively demyelinating, inactive demyelinated lesions, and in remyelinating lesions, Shh-IR was detected in macrophages, endothelium, and astrocytes. Shh-IR in axons was exclusively present in remyelinating lesions. Although the exact molecular mechanisms of the Shh-signaling pathway in experimental autoimmune encephalomyelitis are yet to be determined, our findings may imply a role of Shh signaling in facilitating remyelination. PMID- 15892299 TI - MLC1: a novel protein in distal astroglial processes. AB - Megaloencephalic leukoencephalopathy with subcortical cysts (MLC) is a progressive cerebral white matter disease in children caused by mutations in the MLC1 gene. This disease is histopathologically characterized by myelin splitting and intramyelinic vacuole formation. MLC1 encodes a novel protein, MLC1, which is mainly expressed in the brain and leukocytes. The function is unknown, although a transport function has been suggested. In this article, we provide experimental data addressing the membrane topology and cellular localization of MLC1. We show that MLC1 contains an even number of transmembrane domains, supporting the possible transport function of MLC1. We demonstrate that MLC1 is specifically expressed in distal astroglial processes in perivascular, subependymal, and subpial regions. This localization suggests a role for MLC1 in a transport process across the blood-brain and brain-cerebrospinal fluid barriers. Astrocyte functions have long been debated. It is becoming increasingly clear that these cells are of fundamental importance in maintaining the structural and functional integrity of neural tissue. Elucidation of the function of MLC1 will contribute to a better understanding of not only the pathophysiology of the disease, but also the role of astrocytes in normal neural tissue. PMID- 15892300 TI - Neuropathologic, biochemical, and molecular characterization of the frontotemporal dementias. AB - The frontotemporal dementias (FTDs) are a heterogeneous group of neurodegenerative disorders that are characterized clinically by dementia, personality changes, language impairment, and occasionally extrapyramidal movement disorders. Historically, the diagnosis and classification of FTDs has been fraught with difficulties, especially with regard to establishing a consensus on the neuropathologic diagnosis. Recently, an international group of scientists participated in a consensus conference to develop such neuropathologic criteria. They recommended a diagnostic classification scheme that incorporated a biochemical analysis of the insoluble tau isoform composition, as well as ubiquitin immunohistochemistry. The use and reliability of this classification system has yet to be examined. In this study, we evaluated 21 cases of FTD. Using traditional histochemical stains and tau protein and ubiquitin immunohistochemistry, we separated each case into one of the following categories: classic Pick disease (PiD; n = 7), corticobasal degeneration (CBD; n = 5), dementia lacking distinctive histopathologic features (DLDH; n = 4), progressive supranuclear palsy (PSP; n = 2), frontotemporal lobar degeneration with motor neuron disease or motor neuron disease-type inclusions (FTLD-MND/MNI; n = 2), and neurofibrillary tangle dementia (NFTD; n = 1). Additionally, we independently categorized each case by the insoluble tau isoform pattern, including 3R (n = 5), 4R (n = 7), 3R/4R (n = 3), and no insoluble tau (n = 6). As suggested by the proposed diagnostic scheme, we found that the insoluble tau isoform patterns correlated strongly with the independently derived histopathologic diagnoses (p < 0.001). The data show that cases containing predominantly 3R tau were classic PiD (100%). Cases with predominantly 4R tau were either CBD (71%) or PSP (29%). Cases with both 3R and 4R tau were either a combination of PiD and Alzheimer disease (67%) or NFTD (33%). Finally, cases with no insoluble tau were either DLDH (67%) or FTLD-MND/MNI (33%). To further characterize these cases, we also performed quantitative Western blots for soluble tau, APOE genotyping, and, in selected cases, tau gene sequencing. We show that soluble tau is reduced in DLDH and FTLD-MND/MNI and that APOE4 is overrepresented in PiD and DLDH. We also identified a new family with the R406W mutation and pathology consistent with NFTD. This study validates the recently proposed diagnostic criteria and forms a framework for further refinement of this classification scheme. PMID- 15892301 TI - Is medicine moving uptown? PMID- 15892303 TI - The integrated future. PMID- 15892302 TI - The semantics of disability. PMID- 15892304 TI - Commitment to excellence. PMID- 15892305 TI - Evaluating the options. The pros and cons of dependent mobility systems. PMID- 15892306 TI - Adding to the athlete's rehab arsenal. PMID- 15892307 TI - Age and water do mix. PMID- 15892308 TI - Choosing the right cushion. PMID- 15892309 TI - Pulling the plug on pain. PMID- 15892310 TI - The fall factor. PMID- 15892311 TI - Bracing vs tape. PMID- 15892312 TI - CMS strikes again. PMID- 15892313 TI - As the focus narrows. PMID- 15892314 TI - Surviving and thriving. PMID- 15892315 TI - Increasing amount of concern expressed about the future leadership of the Canadian health services system. PMID- 15892316 TI - "Thank you" isn't enough. PMID- 15892317 TI - Identification of best practices in the delivery of patient food services through public/private sector partnerships. AB - This article describes the evolution of public/private sector partnership in patient food service delivery and identifies skills required by executives to manage partnerships effectively, features that may be generalized to other areas; and the importance of labour relations. Site visits conducted across Canada demonstrate that when partners understand each other's objectives, commit to providing strong leadership, and create meaningful labour relations and communications strategies, partnerships can be successful. PMID- 15892318 TI - Case management approaches for in-home care. AB - Restructuring healthcare delivery systems within limited budgets presents multiple management challenges. As hospitals have downsized, the demand for in home services has increased. The involvement of multiple provider agencies and professionals has heightened duplication and fragmentation of services and has complicated the need to develop client-focused, evidence-based approaches. In response to these challenges, community-based case management has burgeoned. PMID- 15892319 TI - Information systems for healthcare: why haven't more success. The top 15 reasons. AB - Over the last number of years, I have often been asked: "Why haven't we had more success in implementing Information Technology (IT) in Healthcare?" Unfortunately, there is no simple answer to this question. The answer is usually heavily dependent on several factors that "define" the specific implementation in question--consequently, the answer is one comprised of a number of interrelated factors or components. In order to facilitate this answer process, this paper attempts to identify these individual answer components. At the very least, this will help simplify the process of answering future questions by referring to the components outlined herein. At most, in addition to providing a reference compendium for others, it will assist in increasing the solution implementation success rate by exploring the problem definition in detail: The first step in solving a problem is to have it fully articulated. PMID- 15892320 TI - Volunteering a rung on the career ladder? PMID- 15892321 TI - Integrated systems to reduce length of stay for knee and hip joint replacement surgeries. AB - To reduce length of stay while maintaining quality of care, St. Joseph's Hospital, Hamilton, ON implemented a care path with three discharge options. Two of these discharge options were early discharge to integrated community services. Patients meeting early discharge criteria are discharged home five days post operatively with follow-up by home care nursing and physiotherapy. Otherwise, patients are discharged on day four to a multi-disciplinary rehabilitation unit at a separate facility. Patients requiring acute medical services for complications or co-morbidity stay in the acute care hospital. A prospective cohort evaluation showed no difference in complications and similar functional outcomes for the three discharge options. PMID- 15892322 TI - Working to promote and maintain a safe and secure environment in a hospital setting. AB - This article examines how one hospital developed an approach to creating a safer and more secure environment for staff and patients. A cross-functional task group promoted a review of existing practices and processes related to the issue of violence in the workplace. The task group made recommendations for improving prevention, response, and the reporting of violent incidents. Results to date indicate that this approach is working to meet the goal of a safer and more secure environment. PMID- 15892323 TI - Regionalization in PEI seven years later: integrated health and social services bring teamwork and improved care [interviewed by Matthew D. Pavelich]. PMID- 15892324 TI - Rebuilding organizational culture in the wake of a merger. AB - More and more, corporate culture is being recognized as an important determinant of organizational effectiveness. This article describes how the Centre for Addiction and Mental Health proactively focused on rebuilding its culture after a mandated merger. The long-term effects of this effort need to be monitored. However, commitment is strong to the ideas articulated in the Statement of Desired Culture that was developed by a staff task force and approved by the Centre's Board. PMID- 15892325 TI - Impact of restructuring scale: an instrument to measure effects of hospital restructuring. AB - As restructuring and downsizing occur throughout the workplace, many individuals are either losing their jobs or experiencing job insecurity. The study of downsizing is particularly important within the healthcare system where hospital mergers and closings, and severe cutbacks, have dramatically reduced healthcare services. Since nurses are the largest group employed by hospitals, they are the most likely to be affected by recent cutbacks. All this leads to the conclusion that a measure of the impact of restructuring is needed. This study reports on the Impact of Restructuring Scale--a new scale containing acceptable psychometric properties--that quantifies the effects of restructuring on organizations and individuals. The scale was applied to a sample of 1,363 nurses employed in hospitals undergoing restructuring and downsizing. The nurses returned a self report questionnaire in which they reported their reactions to hospital restructuring and to specific job stressors. PMID- 15892326 TI - Redefining leadership responsibilities following organizational redesign. AB - A redefinition of leadership roles was necessitated by the adoption of program management, which accompanied the merger of two large academic hospitals, each of which consisted of two sites. Directors of professional departments were removed when staff were deployed to programs. Program directors were selected to manage the new programs, and profession leaders were identified to monitor standards of practice. The roles and accountabilities of the new program and profession leaders needed to be clarified for incumbents and other hospital staff. This article describes the process undertaken to clarify those responsibilities. The resulting Guide outlines 14 specific and collaborative responsibilities of program and profession leaders. Plans for review and future development of the Guide are discussed. PMID- 15892327 TI - Employment contracts: what to look for from the point of view of both the employer and employee. PMID- 15892328 TI - The increasing reality of using the internet for electronic health records. PMID- 15892329 TI - Prioritizing resource allocation for clinical enhancement: a participative methodology. AB - The allocation of hospital funding for new and expanded clinical programs can be a difficult but most important task to deal with during the development of the operating plan (budget). Like many other hospitals, York Central Hospital has struggled with this task each year. In order to address this challenge, the hospital has successfully designed and implemented a prioritization process that includes a standardized program proposal and peer evaluation. The process is grounded in the hospital's vision and strategic directions and built on a culture of evidence-based practice. PMID- 15892330 TI - Healthcare: the new economy's last frontier? AB - The New Economy is transforming large parts of the business world and the public sector. New Internet technologies are being used to increase efficiency and add value. But the most distinctive contribution of the Internet is the added reach that it is giving to producers and service providers who had previously been confined by geography and by travel time and costs. Now they are able to connect with much larger markets and reach more communities and do so with a much greater immediacy. The Internet is also facilitating much more responsive and tailored services/products. This is giving rise to new and expanded demand, as these Internet-enabled services become more attractive to buyers/consumers. PMID- 15892331 TI - [Screening reduced mortality in abdominal aortic aneurysm]. PMID- 15892332 TI - [Cowpox virus: a relevant etiology in Denmark?]. PMID- 15892333 TI - [Cerebral palsy--what is the influence of genetic factors?]. PMID- 15892334 TI - [Social, linguistic and cognitive development in children with Angelman's syndrome]. PMID- 15892335 TI - [Cognitive function and renewal of driving licences for persons older than 70 years]. PMID- 15892336 TI - [USEUMEE--a comparison of the Danish and American health care systems]. PMID- 15892337 TI - [Ultrasound scanning of abdominal aortic aneurysms. Interobserver variability with and without the use of spatial compound imaging and comparison with computer tomography]. PMID- 15892338 TI - [Screening reduced abdominal aortic aneurysm mortality--secondary publication. Results from a Danish randomized screening trial]. PMID- 15892339 TI - [The fetal erythroblast is not the optimal target for non-invasive prenatal diagnosis--secondary publication]. PMID- 15892340 TI - [Detection of cowpox virus in Denmark]. PMID- 15892341 TI - [Systemic vascular leak syndrome]. PMID- 15892342 TI - [Borrelia lymphocytoma in the breast]. PMID- 15892343 TI - [Picture of the month: contact dermatitis]. PMID- 15892344 TI - [Daily dosage of 10 mg vitamin D to immigrants wearing well-covering dark clothes is not sufficient]. PMID- 15892345 TI - [Mesalamine in the treatment of Crohn disease?]. PMID- 15892346 TI - [Remote drug-review--a temporary solution. Introduce a more expedient organization in geriatric residential facilities!]. PMID- 15892347 TI - [Unexpected findings--sunlight can protect against cancer]. PMID- 15892348 TI - [Remote drug-review for better use of pharmaceuticals among the elderly]. AB - During a decade various actions have been taken to decrease the use of inappropriate medication for elderly patients in Sweden. The Swedish Health Department has recently compiled guidelines for inappropriate medication that can be used to evaluate medication in nursing homes for elderly. At the beginning of our project we assessed the medication of persons living in 21 nursing homes in the northeastern district of the county Skane and based on this analysis we decided that one of the actions to take were drug-reviews. For a period of two years an intensified effort was made to improve medical treatment in nursing homes in the district. Seventeen nursing homes, with 181 patients, were willing to participate. The patients' current medications, state of health (including blood pressure and weight) and known diagnosis were evaluated by a pharmacist and a geriatrician. A specific written recommendation on how to modify drug-usage in these patients were sent to the nursing homes. Decisions were then made by the doctors in charge of the patients on which adjustments to make. At the end of the project 127 patients were reevaluated concerning quality and quantity of medication. A comparison was made before and after the drug-review. Positive results were obtained as well as a lower drug cost. An assessment of the patients' state of health, before and after drug-review, showed a tendency towards improvement. We compared our results with those of a Swedish project that used a different method of drug-review with a visiting pharmacist instead of a written recommendation. The changes in the patients' medication before and after drug-review were the same. PMID- 15892349 TI - [Electromagnetic therapy of circulatory disorders in the lower extremities. Promising advertisements in daily press are not scientifically documented]. PMID- 15892350 TI - [Claudication and pulmonary embolism can be caused by venous aneurysm. A case report illustrates difficulties with this unusual diagnosis]. AB - A 19 year old male patient is described with a large popliteal venous aneurysm causing pulmonary embolism and intermittent claudication. It was resected and replaced with a saphenous vein graft. PMID- 15892351 TI - [Notice to quit because of moonlighting--a simulated case discussion]. PMID- 15892352 TI - [Non-reliable marketing of TrioBe]. PMID- 15892353 TI - [Sweden has much to learn from English health care]. PMID- 15892354 TI - [Adverse effects of sick leave]. PMID- 15892355 TI - [Computer registries not only for the good. Don't forget patient integrity!]. PMID- 15892356 TI - [Health services and the most vulnerable persons I: Not providing care for the homeless is against the law]. PMID- 15892357 TI - [Safer health effects of raisins than wine!]. PMID- 15892358 TI - [Are dog bites never to be sutured?]. PMID- 15892359 TI - Self-curing, self-etching adhesive cement systems. AB - PURPOSE: To investigate the influence of the acid-base reaction between acidic monomers and amines on the polymerization behavior of self-etching, self-curing adhesives, determine the effect of the application mode on the shear bond strength and morphology, and elucidate the adhesion performance of such systems by shear bond strength measurements. MATERIALS AND METHODS: The amine redox initiator system N,N-di(2-hydroxyethyl)p-toluene (DEPT)/dibenzoyl peroxide (BPO) was selected to investigate the influence of the amine-base reaction on polymerization behavior. The PKa value of DEPT hydrochloride was measured by titration with NaOH. The influence of the pH value and DEPT concentration on the polymerization rate of methacrylates was investigated by exotherm time measurements. Three different application protocols of Multilink Primer (Ivoclar Vivadent) and Panavia 21 ED-Primer (Kuraray) were tested, 15 s passive vs 15 s agitation vs 60 s passive in combination with the corresponding resin luting material. The effects of these three application protocols were evaluated and monitored by both shear bond strength tests and SEM characterization of the surface morphology. The adhesion potential of these self-etching, self-curing luting systems was compared on enamel and dentin both directly after application and after 24 h. RESULTS: The pKa of DEPT-HCl is 4.45. The polymerization rate of the DEPT-containing, self-etching, self-curing adhesive system is highly influenced by both the amine concentration and the pH value. In the case of Multilink, agitation of the primer mixture for 15 s, especially on dentin, resulted in a higher bond strength and a more pronounced removal of the smear layer. Multilink resulted in statistically higher bond strengths (p < 0.05) than Panavia 21 for both the enamel and dentin directly after application and for the dentin after 24 h. CONCLUSION: Radical polymerization initiators used in self curing systems are strongly adversely affected by acidic monomers incorporated in self-etching adhesives. However, if there is a good adjustment of these components and effects of adhesive application are taken in account, high bond strengths can be achieved. PMID- 15892360 TI - Fluorescence microscopy for the evaluation of the margins of Class V restorations in vitro. AB - PURPOSE: First, to elucidate the cause of the appearance of "white lines" in Class V restorations as seen under the light microscope and their enhancement when using a fluorescent microscope (FM); second, to compare the results of the FM quantitative marginal analysis to those of the SEM quantitative marginal analysis. MATERIALS AND METHODS: Thirty-two standardized Class V fillings, with half of the preparation in dentin and half in enamel, were placed in 16 human caries-free premolars on the buccal and lingual surface using 4 adhesive systems (Clearfil SE, Syntac, 2 experimental self-etching adhesives) and the composite Tetric Ceram. All teeth were connected to a device containing horse serum to simulate dentinal fluid and subjected to thermomechanical loading (1,200,000 cycles at 49 N, 3000 cycles at 5 degrees C/55 degrees C). The restoration margins of 24 fillings were directly evaluated with FM--after the fabrication of replicas -and with SEM. Eight fillings were used to elucidate the cause of the fluorescent signal, by examining them with FM and different filters under dry and moist conditions, and also with dark field microscopy and CLSM. Further, two Class V fillings with the same composite but without the fluorescent substances were made and evaluated. Selected specimens were cut in the bucco-oral direction to analyze the restorative interface on section replicas with SEM. To detect differences between FM and SEM, the Wilcoxon test was performed (p < 0.05). To evaluate the degree of agreement between the two evaluation methods, a linear regression analysis was performed and the Spearman correlation coeffcient was calculated. RESULTS: The evaluation of Class V restorative margins by SEM, dark field microscopy, and CLSM revealed that the phenomenon seems to be an optical effect caused by the debonding of the composite from the underlying substrate creating another refractive environment when air penetrates into the gap. The effect depends on the geometry of the cavity design, the absence of moisture, and the use of a fluorescent composite material. The light source that produced the best visible results covered both ultraviolet and the blue part of white light (350 to 460 nm), producing light green lines and green areas. Those green or white areas as seen with the light microscope were related to gaps in the interface between tooth substance and composite. With the exception of one of the experimental self etching adhesives, the mean percentage of continuous margin of the 4 groups showed no statistically signifcant dfference for the two test methods. Linear regression analysis revealed a good correlation between the two test methods for dentin margins and a moderate correlation for enamel margins. CONCLUSION: Fluorescence microscopy seems to be a useful tool for evaluating dentinal and enamel margins of Class V restorations in vitro. PMID- 15892361 TI - Effect of ozone on enamel and dentin bond strength. AB - PURPOSE: To evaluate the influence of direct high-dose gaseous ozone application (2100 ppm) on dentin and enamel shear bond strength. MATERIALS AND METHODS: Ten bovine enamel and dentin samples per group were pretreated as follows: (I) ozone application (Healozone, KaVo) for 60 s alone or (II) with subsequent application of a fluoride- and xylitol-containing antioxidant (liquid reductant), (III) light activated bleaching with 35% hydrogen peroxide for 5 min serving as negative control (Hi-Lite, Shofu), and (IV) untreated enamel and dentin (positive control). Specimens were bonded with a functional 3-step adhesive system (Syntac Classic, Ivoclar Vivadent) and restored with a composite (Tetric Ceram, Ivoclar Vivadent) according to the Ultradent method. After storage in water at 37 degrees C for 24 h, shear bond strength was measured using a Zwick universal testing machine. Data were analyzed using ANOVA and Scheffe's post hoc analysis. RESULTS: In concordance with the existing literature, bleaching resulted in significantly decreased bond strength (p < 0.05) on enamel specimens. No decrease in shear bond strength was detected for ozone-pretreated specimens compared to untreated controls. CONCLUSION: Despite a possible retention of surface and subsurface oxide-related substances during high-dose ozone application, shear bond strength was not impaired. Thus, adhesive restoration placement should be possible immediately after ozone application for cavity disinfection. PMID- 15892362 TI - In vitro evaluation of wall-to-wall adaptation of a self-adhesive resin cement used for luting gold and ceramic inlays. AB - PURPOSE: This in vitro study evaluated the wall-to-wall adaptation of a new self adhesive resin-based cement (RelyX Unicem) in comparison with that of other cements when luting gold and porcelain inlays in standardized Class II cavities in extracted teeth. MATERIALS AND METHODS: In each experimental group (n = 10), a different combination of inlay and luting material was tested. Group 1: Porcelain Empress II (Ell) and RelyX Unicem (U); group 2: Ell and resin-based cement Variolink II in combination with primer and bonding Excite DSC; group 3: gold inlays (G) and U; group 4: G and Harvard zinc-oxy-phosphate cement; group 4: G and glass-ionomer cement Fuji Cem. After storage and thermocycling, microleakage testing was carried out and dye penetration was examined at the occlusal and cervical margins of each inlay. The differences in microleakage score were tested for statistical significance first comparing all groups, then pooling the groups for inlay material (Kruskal-Wallis nonparametric ANOVA and Mann-Whitney U test, p < 0.05). SEM observations of the tooth/cement/restoration interfaces were also made in each group. RESULTS: Harvard cement had the highest microleakage. The sealing ability exhibited by RelyX Unicem was satisfactory with both gold and porcelain inlays, and comparable to that of Fuji Cem and Variolink II. CONCLUSION: RelyX Unicem achieved an adequate seal on both enamel and dentin when used to lute in vitro gold and porcelain inlays. PMID- 15892363 TI - Evaluation of varied repair protocols applied to aged composite resin. AB - PURPOSE: This paper aims to examine varied surface treatments combined with adhesive bonding to ascertain the highest bond strength of the composite resin repair. MATERIALS AND METHODS: A total of 100 specimens was made using an increment technique. The prepared specimens were aged for 20 days in 37 degrees +/- 20 degrees C water and then kept for 24 h at room temperature. The samples were separated into 5 groups of 20, and each group then received a different surface treatment (control group, phosphoric acid, diamond bur, air abrasion, silane primer combined with a diamond bur treatment). The effect of each surface treatment on the bond strength was determined by a shear bond test. RESULTS: Specimens that received surface treatment with a diamond bur exhibited higher shear bond strengths than any of the other repaired groups (p < 0.05). Those treated with air abrasion had the second highest shear bond strengths, but without a statistically significant difference when compared to the control group (p > 0.05). Specimens treated with phosphoric acid showed virtually no difference compared to the specimens of the control group (p > 0.05). CONCLUSION: Statistical analysis revealed that surface treatment with a diamond bur results in higher shear bond strength than other tested methods. PMID- 15892364 TI - SEM analysis of internal adaptation of adhesive restorations after contamination with saliva. AB - PURPOSE: This study tested the null hypothesis that different treatments of saliva-contaminated substrate would not affect microgap formation at the dentin walls of bonded restorations. MATERIALS AND METHODS: Forty freshly extracted human molars received standardized Class V preparations on buccal and lingual surfaces. The specimens were assigned to four experimental groups (n = 20): [G1] no contamination (control group), [G2] saliva contamination (10 s) after etching followed by 5 s air stream; [G3] saliva contamination after etching and rinsed for 10 s; and [G4] re-etching for 10 s after saliva contamination. All specimens were restored with a one-bottle adhesive (Single Bond, 3M ESPE) and microhybrid composite resin (Filtek Z250, 3M ESPE) according to the manufacturer's instructions. The specimens were thermocycled, sectioned through the center of the restoration, and then processed for SEM. Microgaps were measured at the axial wall at 1500X magnification. The data were submitted to Kruskal-Wallis nonparametric statistical analysis at p < 0.05. RESULTS: The data revealed that different groups resulted in a statistically significant difference (p < 0.01) in gap formation. Air drying [G2] and rinsing [G3] the saliva-contaminated dentin resulted in similar microgap values (p > 0.05). However, re-etching the dentin after saliva contamination [G4] increased microgap formation (p < 0.05) when compared with the groups G1 and G2. Although air drying and rinsing produced results comparable to noncontaminated dentin, the presence of microgaps was not completely eliminated. CONCLUSION: Contaminated saliva did not prevent hybrid layer formation; however, it did reduce the adaptation of the restorative material to bonded surfaces. PMID- 15892365 TI - Shear bond strength of one 4th and two 7th generation bonding agents when used by operators with different bonding experience. AB - PURPOSE: To test the hypothesis that simpler-to-use dentin adhesives (7th generation adhesives) perform better when used by inexperienced operators than do the more complex 4th-generation adhesives. MATERIALS AND METHODS: Six operators with no previous experience regarding dental adhesives, three residents in pediatric dentistry, and three experienced dentists used one 4th-generation dentin adhesive (MA) and two 7th-generation adhesives (MB and MC). With each adhesive, each operator performed 6 bondings to enamel and 6 bondings to dentin. After 24 h of storage in water at 37 degrees C, the shear bond strength was determined. RESULTS: The pooled results showed that the bond strength to dentin was higher (p < 0.05) than that to enamel (dentin mean value = 14.0 MPa, SD = 9.1 MPa; enamel mean value = 11.9 MPa, SD = 7.1 MPa), and that adhesive MA performed better (p < 0.05) than adhesives MB and MC (inexperienced operators = 16.2 +/- 10.9 MPa; residents in pedodontics = 12.0 +/- 6.8 MPa, and experienced operators = 10.7 +/- 4.8 MPa). Of the two 7th-generation adhesives, one performed better on enamel than on dentin, while the other 7th-generation adhesive performed better on dentin than on enamel. Regarding operators, there were large individual variations. The inexperienced bonders performed as well as the residents (p > 0.05), while the experienced operators performed best (p < 0.05). CONCLUSION: In general, the tested 4th-generation adhesive performed better than the 7th generation adhesives. Even inexperienced operators performed better with the more difficult-to-use 4th-generation adhesives than with the 7th-generation adhesives. Dentin adhesion was stronger than enamel adhesion. PMID- 15892366 TI - Two-year clinical evaluation of One-Up Bond F in noncarious cervical lesions. AB - PURPOSE: To evaluate the retention of restorations placed in non-undercut, noncarious cervical lesions using One-Up Bond F and Palfique Estelite resin composite. MATERIALS AND METHODS: Fifty-one non-undercut, noncarious cervical lesions were restored with One-Up Bond F and Palfique Estelite in 15 patients (mean age 57.7 years). Teeth were cleaned with pumice and water and restored following the manufacturer's instructions. Restorations were evaluated at 6 months, 1 and 2 years for presence or absence, and for the degree of marginal staining using standardized colour photographs for comparison. RESULTS: At 1 year, 42 restorations were evaluated, and all were intact. Slight marginal staining was observed in three restorations. At 2 years, 37 restorations were evaluated from 11 patients, and all were intact. Marginal staining was observed around nine restorations. ANOVA for marginal staining showed a significant increase of staining at both the enamel and cervical margins over the 2 years of the study. No caries was detected in association with any restoration. CONCLUSION: Although a number of restorations could not be evaluated, the retention of 100% at 2 years showed very good promise for the treatment of noncarious cervical lesions. The marginal staining was limited to a small number of patients. PMID- 15892367 TI - Building custom shells for conservative tooth reconstruction: an elegant strategy. AB - This article, focused around case presentations, follows the natural progression/exploration of a single idea about dealing with the Class II (approximal lesion) composite restoration. The gradual development of this idea and the integration of a concept from one case to the next may have certain educational value. Hopefully it will show the potential for further experimentation and inspire a fresh mentality in using and handling the main line of conservative restorative materials today, ie, resin composites. PMID- 15892368 TI - Invalid measurement validity. PMID- 15892369 TI - Neonatal encephalopathy: etiology and outcome. PMID- 15892370 TI - Cerebral palsy following term newborn encephalopathy: a population-based study. AB - Cerebral palsy (CP) can occur in term infants with or without preceding newborn encephalopathy. We compared the type and severity of CP and associated disability in these two groups. Participants from a population-based case-control study of term newborn encephalopathy were followed up for 6 years and linked to the Western Australian Cerebral Palsy Register. The remaining term infants with CP for the same period were also identified from the Cerebral Palsy Register. 13% of neonatal survivors of term newborn encephalopathy had CP, a rate of 116 per 1000 term live births. Overall, 24% of term infants with CP followed newborn encephalopathy. CP following newborn encephalopathy was more likely to: affect males (72% vs 56%); be severe (47% vs 25%); and be of spastic quadriplegia or dyskinetic types. Cognitive impairment was more common (75% vs 43%) and severe (41% vs 16%), as was epilepsy (53% vs 29%) in survivors of encephalopathy. These children were also more likely to: be non-verbal (47% vs 22%); have a severe composite disability score (47% vs 26%); and die between time of diagnosis of CP and age 6 years (5-year cumulative mortality 19% vs 5%). Children born at term who develop CP following newborn encephalopathy have a poorer prognosis than those with CP who were not encephalopathic in the first week of life. PMID- 15892371 TI - Neonatal feeding performance as a predictor of neurodevelopmental outcome at 18 months. AB - This study aimed to determine whether neonatal feeding performance can predict the neurodevelopmental outcome of infants at 18 months of age. We measured the expression and sucking pressures of 65 infants (32 males and 33 females, mean gestational age 37.8 weeks [SD 0.5]; range 35.1 to 42.7 weeks and mean birthweight 2722g [SD 92]) with feeding problems and assessed their neurodevelopmental outcome at 18 months of age. Their diagnoses varied from mild asphyxia and transient tachypnea to Chiari malformation. A neurological examination was performed at 40 to 42 weeks postmenstrual age by means of an Amiel-Tison examination. Feeding performance at 1 and 2 weeks after initiation of oral feeding was divided into four classes: class 1, no suction and weak expression; class 2, arrhythmic alternation of expression/suction and weak pressures; class 3, rhythmic alternation, but weak pressures; and class 4, rhythmic alternation with normal pressures. Neurodevelopmental outcome was evaluated with the Bayley Scales of Infant Development-II and was divided into four categories: severe disability, moderate delay, minor delay, and normal. We examined the brain ultrasound on the day of feeding assessment, and compared the prognostic value of ultrasound and feeding performance. There was a significant correlation between feeding assessment and neurodevelopmental outcome at 18 months (p < 0.001). Improvements of feeding pattern at the second evaluation resulted in better neurodevelopmental outcome. The sensitivity and specificity of feeding assessment were higher than those of ultrasound assessment. Neonatal feeding performance is, therefore, of prognostic value in detecting future developmental problems. PMID- 15892372 TI - Visual and spatial long-term memory: differential pattern of impairments in Williams and Down syndromes. AB - This purpose of this study was to investigate visual-object and visual-spatial long-term memory (LTM) abilities in individuals with Williams syndrome (WS) and Down syndrome (DS). Four groups comprised of 15 participants were included: WS group (10 males) with a mean chronological age (CA) of 18 years 5 months, SD 6 years 4 months, and mean mental age (MA) of 6 years 8 months, SD 1 year 5 months; WS control group (eight males) comprised of typically developing children (CA mean 6y 7mo, SD 8mo); DS group, (10 males, CA mean 16y 5mo, SD 5y 10mo; MA mean 5y 4mo, SD 8mo); and DS control group (seven males) formed by typically developing children (CA mean 5y 6mo, SD 7mo). In the WS and DS groups mental age and IQ were evaluated with the Form L-M of the Stanford-Binet Intelligence Scale. Results showed that individuals with WS showed decreased learning of visual spatial material but substantially typical learning of visual-object patterns as compared to a group of mental-age-matched typically developing children. Individuals with DS showed the opposite profile, i.e. typical learning of visual spatial sequences but impaired learning of visual-object patterns. These results, showing an interesting double dissociation between these two genetic syndromes in the learning of visual-object patterns as opposed to visual-spatial data, support the interpretation of learning disability* as a heterogeneous condition, characterized by potentially very different qualitative profiles of cognitive impairment. PMID- 15892373 TI - Development of postural adjustments in sitting position during the first half year of life. AB - Little is known about the development of postural adjustments during early ontogeny. We examined postural adjustments due to sudden perturbations during sitting in 40 healthy term infants (28 males, 12 females) assessed in groups of eight at 1, 2, 3, 4, and 5 months of age. Surface electromyograms of neck, trunk, and leg muscles were recorded while the infants were exposed to a random series of horizontal forward and backward displacements of the surface of support. Video recordings of spontaneous motor behaviour were analyzed. For part of the analyses, previously collected data on 26 infants aged 6 to 10 months were included. In general, postural adjustments at all ages were direction specific and showed large variation. Within the variation developmental changes could be observed, revealing a transient decrease in postural activity at 3 months of age. After this transition, direction-specific postural activity was correlated with spontaneous motor behaviour. This was true, in particular, for dorsal postural activity. The clinical relevance of these findings is discussed. PMID- 15892374 TI - Gait adaptation of children with cerebral palsy compared with control children when stepping over an obstacle. AB - This study investigated the adaptive gait pattern in obstacle clearance in 12 normally developing (ND) children (six males, six females; mean age 10y 2mo, SD 10mo) and 12 children who had spastic diplegic cerebral palsy (CP) and who were independent ambulators (10 males, two females; mean age 13y 5mo, SD 2y 7mo). Children in both groups had 8 to 12 years of walking experience. They walked up to and crossed over obstacles of a height of 0%, 10%, and 20% of their leg length. Kinematic aspects of the three-dimensional leg movements were captured with the Vicon system and analyzed with one-way analysis of variance with repeated measure. Both groups increased foot clearance for higher obstacles. Children with CP were slower in approach and crossing speed, and used a wider base of support than the ND children when the obstacle height was increased. Results suggest that motor problems in obstacle clearance in children with CP lie in the implicit process of motor control, i.e. torque dynamics, not the explicit process, i.e. movement-match with the environmental constraints. PMID- 15892375 TI - Neuromuscular activation and motor-unit firing characteristics in cerebral palsy. AB - Muscle strength, neuromuscular activation, and motor-unit firing characteristics (firing rate, recruitment, and short-term synchronization) were assessed during voluntary contractions of the medial gastrocnemius (GAS) and tibialis anterior (TA) muscles of 10 participants with spastic diplegic or hemiplegic cerebral palsy (CP). The participants (six females, four males; age range 6 to 37y) walked with equinus gait at Gross Motor Function Classification System levels II to III. These were compared with 10 age-matched controls (five females; age range 7 to 35y). Neuromuscular activation was estimated by the ratio of surface electromyogram amplitude to M-wave amplitude elicited by supramaximal electrical nerve stimulation. Participants with CP produced significantly less torque (normalized by leg length) compared with controls (TA: mean 2.3, SD 1.6 vs mean 8.9, SD 3.4Nm/m; GAS mean 13.7, SD 7.1 vs mean 28.6, SD 5.1Nm/m, p < 0.001). Neuromuscular activation during maximum voluntary contraction was significantly reduced in the participants with CP compared with controls (mean 2.4, SD 1.5 vs mean 9.7, SD 2.7Nm/m for TA; mean 1.04, SD 0.41 vs mean 3.1, SD 1.2Nm/m for GAS, p < 0.001). When compared at the same submaximal level of neuromuscular activation, motor-unit recruitment and firing rates were not different between the groups, although short-term synchronization in TA was reduced in the participants with CP. These data indicate that weakness, known to be an important component of the motor deficit in CP, has a strong central component. Although the relation between recruitment and firing rate remained substantially intact at the low and moderate force contractions tested, results suggest that the participants with CP were unable to recruit higher threshold motor units or to drive lower threshold motor units to higher firing rates. PMID- 15892376 TI - Muscle force generation and force control of finger movements in children with spastic hemiplegia during isometric tasks. AB - Force control ability was investigated in 10 males and 10 females, between 5 and 15 years old with spastic hemiplegia (mild and moderate hand dysfunction), and an aged-matched control group (eight males, 12 females). An isometric force production task at five different levels of maximum voluntary contraction (MVC) was performed. Results showed that MVC generated with the affected hand (AH) was only one-third of that generated by the non-affected hand (NAH; p < 0.001), time to peak was almost twofold at the highest force level (p < 0.001), and the coefficient of variation was twice as high (p < 0.001). Results for the NAH did not differ from those of the control children. Correlations between clinical and experimental variables were significant for the relation between Ashworth score for elbow flexors, MVC and variability at the highest force level. In conclusion, the findings for the AH suggests that strength training should be considered for agonist spastic muscles. PMID- 15892377 TI - Ring chromosome 20 syndrome with intractable epilepsy. AB - Ring chromosome 20 (r[20]) syndrome is characterized by mild to moderate learning disability*, behavioural disorders, epilepsy, and various dysmorphic features. Although still considered rare, r (20) syndrome is being increasingly diagnosed. More than 30 cases have been described in the literature since 1976. Here we report an additional case of a 14-year-old male with r (20). He had moderate to severe learning disability and epileptic seizures manifesting at about 18 months of age. During the 13 years' follow-up period he showed intractable epileptic seizures, behavioural disorders, and mild dysmorphological features including microcephaly, strabismus, micrognathia, down-slanting eyelids, and ear abnormalities. Frequent episodes of atypical absence or non-convulsive status associated with electroencephalogram changes were seen in follow-up. He was treated with several classical and new antiepileptic drugs, including intravenous immunoglobulin, corticotropin, and vagal nerve stimulation, with unsuccessful control of seizures. Finally, surgical treatment (corpus callosotomy) was performed at the age of 13 years; severity of tonic seizures was diminished, but frequency was unchanged. Although his behavioural problems, e.g. hyperactivity, were mild in early childhood they became more severe when he was 11 years old. Aggressiveness, compulsiveness with self-injury, and panic attacks developed at the age of 13 years, and were more pronounced after callosotomy. This case report provides the first description of deterioration in psychological situation in patients with r(20) intractable epilepsy. The patient was diagnosed with r(20) syndrome after 13 years of clinical follow-up. Karyotype analysis should, therefore, be performed in every patient with intractable epilepsy of unknown aetiology. PMID- 15892378 TI - Assessing pulmonary consequences of dysphagia in children with neurological disabilities: when to intervene? PMID- 15892379 TI - Interpretation and implementation of oxygen uptake kinetics studies in children with spastic cerebral palsy. AB - From the scant data available, it seems that children with CP have a low maximum Vo2, a high O2 cost of submaximal exercise, and inefficient gait mechanics characterized by high levels of co-contraction. Despite the non-steady-state nature of habitual activity, there seem to be no published data focusing on Vo2 kinetics in children with CP Before measuring Vo2 kinetics, the noise content of the data needs to be examined because this has important implications for estimating tau Vo2 values. On the basis of the preliminary findings presented here, the breath-by-breath Vo2 responses of children with CP seem to have a similar noise magnitude to those of typically developing children; however, because they have smaller response amplitudes, the averaging of several repeated trials will be even more important than in typically developing children. Thorough habituation will be required to enable participants to complete the simulated step change exercise protocols that will enable Vo2 kinetics to be quantified. To facilitate appropriate comparisons with control groups on the basis of relative exercise intensities, further work is needed to establish the efficacy of estimating T(Lac) from gas exchange data in children with CP. By measuring Vo2, kinetics in children with CP, it is hoped that we will not only gain valuable insights into their exercise intolerance, but we will also have an important tool for assessing the effects of interventions using submaximal, and therefore less strenuous, exercise. PMID- 15892380 TI - 'Long-term survivor with X-linked myotubular myopathy'. PMID- 15892381 TI - The quest for the global elimination of leprosy. PMID- 15892382 TI - The control of hypertension by use of coconut water and mauby: two tropical food drinks. AB - In this study, the authors investigated the effect of regular consumption of two tropical food drinks, coconut (Cocos nucifera) water and mauby (Colubrina arborescens), on the control of hypertension. Twenty-eight hypertensive subjects were assigned to four equal groups and their systolic and diastolic blood pressures recorded for two weeks before and then for another two weeks while receiving one of four interventions. One group (the control) received bottled drinking water, the second group received coconut water, the third received mauby and the fourth group, a mixture of coconut water and mauby. Significant decreases in the mean systolic blood pressure were observed for 71%, 40% and 43% respectively of the groups receiving the coconut water, mauby and the mixture (p < or = 0.05). For these groups, the respective proportions showing significant decreases in the mean diastolic pressure were 29%, 40% and 57%. For the group receiving the mixture, the largest decreases in mean systolic and mean diastolic pressure were 24 mmHg and 15 mmHg respectively; these were approximately double the largest values seen with the single interventions. PMID- 15892383 TI - Paediatric open heart surgery in Trinidad and Tobago: an example of collaborative care. AB - Between September 1998 and February 2003, 204 children underwent cardiac surgery in Trinidad and Tobago to correct or palliate congenital heart disease. The defects included isolated ventricular or atrial septal defects as well as tetralogy of fallot and coarctation of the aorta. A few patients also had complex cyanotic congenital heart disease. The patients ranged in age from 2 days to 17 years. In some cases, palliative surgery was undertaken as a first step towards complete correction but the majority of patients underwent complete repair. The commonest postoperative complications were haemorrhage and small transient pericardial effusions. The overall mortality rate was < 1%. This paper describes the cardiac lesions, post-operative morbidity and mortality of these patients. PMID- 15892384 TI - Urban Jamaican children's exposure to community violence. AB - Exposure to violence in childhood is associated with aggression in adulthood. The high level of community violence in Jamaica is likely to expose Jamaican children to violence. There has been no detailed study of the exposure of Jamaican children to violence in their daily lives. Some 1674 urban 11-12-year-old children, previously part of a national birth cohort study, completed a questionnaire detailing their exposure to violence as witnesses, victims and aggressors. Their parents completed a socio-economic questionnaire. Jamaican children had high levels of exposure to physical violence. A quarter of the children had witnessed severe acts of physical violence such as robbery, shooting and gang wars, a fifth had been victims of serious threats or robbery and one in every twelve had been stabbed. Children reported being least exposed to sexual violence and to being shot at. Robbery was an almost universal experience affecting children from all schools and socio-economic groups. The single commonest experience as a victim of violence was the loss of a family member or close friend to murder, affecting 36.8% of children. Children's experiences of witnessing violence occurred chiefly in their communities but their personal experiences of violence occurred at school. Boys and children attending primary school had greater exposure to violence as witnesses and victims. Socio-economic status discriminated exposure to physical violence as witnesses but not as victims. Intervention strategies to reduce children's exposure to violence should include community education on the impact of exposure to violence on children, particularly the loss of a significant person, and the development of a range of school-based violence prevention programmes. PMID- 15892385 TI - Beta-haemolytic streptococci in school children 5-15 years of age with an emphasis on rheumatic fever, in the tri-island state of Grenada. AB - The objective of this study is to determine the prevalence of beta-haemolytic streptococci in the pharynx and antibodies to beta-haemolytic streptococci in school children 5-15 years of age in the tri-island state of Grenada Blood samples and throat swabs were obtained from 1388 school children, aged 5-15 years old attending randomly selected schools in each parish of the tri-island state of Grenada. Serum samples were subjected to antistreptolysin o testing (ASOT) while throat swabs were cultured on sheep blood agar plates. The prevalence of positive throat swab was 15.4% (95% CI = 13.4%, 17.2%). The rate was highest in St Patrick (21.8%) and lowest in Carriacou (5.7%). The prevalence of antibodies was 38.6% (95% CI = 37.6%, 42.8%). It was highest in St Patrick (54.4%) and lowest in Petit Martinique (26.8%). In St Patrick, males were significantly more likely than females to have a positive ASOT (p = 0.0084). In St George's, males were significantly more likely than females to have a positive throat culture (p = 0.0172). Thirty-four per cent of the positive cultures were type A, 10% were type C and 56% type G. The data illustrate that there is a high prevalence of beta haemolytic streptococci in school children in certain parishes in Grenada. Public health measures should address prevention and control of beta3-haemolytic streptococcal infection in order to prevent the possible sequelae of this disease. PMID- 15892386 TI - Breastfeeding patterns among six-week-old term infants at the University Hospital of the West Indies. AB - Eighty-nine mothers attending post-natal clinics at the University Hospital of the West Indies were interviewed at six weeks postpartum regarding their breast feeding patterns and problems. Breastfeeding was practised by 97.8% of the mothers, with 29.9% practising exclusive breastfeeding and 70.1% partial breastfeeding. Only two women were solely bottle feeding. The pattern of breastfeeding was not significantly affected by maternal parity, age, education, employment or socioeconomic status. An intention to wean later (at six months) or when the mother felt the baby was 'ready,' was associated with increasing parity, age and further education. Babies who were exclusively breastfed achieved greater weight gain compared with those who partially breastfed but this difference did not achieve statistical significance. Breastfeeding trends appear to have remained stable over the last several years. PMID- 15892387 TI - Neuroendoscopy in Jamaica. AB - This retrospective, descriptive study reviewed the patient profile, disease spectrum, indications for and results of treatment and complications of all 27 patients who had neuroendoscopy at the University Hospital of the West Indies (UHWI) over the three-year period between November 2000, when the service was first introduced, and November 2003. Nineteen (67.9%) were males and eight were females. Their mean age was 27.5 +/- 21.4 years with a range of four months to 70 years. Of the entire group, 20 had hydrocephalus with raised intracranial pressure, of which 15 had endoscopic third ventriculostomy (ETV) for the treatment of obstructive hydrocephalus, using two different techniques for ventricular floor fenestration. There was no demonstrable difference in outcome between the water jet and blunt forceps techniques. Three procedures failed to relieve the hydrocephalus, requiring subsequent ventriculo-peritoneal shunts. Three patients had successful cyst fenestrations. Six patients had endoscope assisted trans-sphenoidal resection for pituitary tumours. There were five complications and no deaths. Mean hospital stay was 18.4 +/- 16.7 days and mean follow-up was 29 weeks. There were no late failures. Neuroendoscopy is the treatment of choice for obstructive hydrocephalus due to aqueduct stenosis or posterior fossa tumours and has the advantage of avoiding shunt related complications. It is safe and effective for the majority of patients and has a significant role in the management of neurosurgical patients in the Caribbean. PMID- 15892388 TI - Surgical complications of cocaine body-packing: a survey of Jamaican hospitals. AB - Body-packing is a common method of smuggling cocaine. Complications requiring surgery do not occur with sufficient frequency to allow any individual surgeon to determine patterns of presentation and the best approach to the conduct of surgery. A survey of all surgical units in Jamaica was conducted. A case was any patient requiring surgery for cocaine body-packing since and including the first reported case in 1987. Seventeen cases were identified. There were 11 cases of bowel obstruction, two of delayed passage of pellets, three of ruptured pellets with cocaine toxicity and one patient panicked and requested surgery. The distal ileum was the commonest site of obstruction in the normal gastrointestinal tract. In all three cases with cocaine poisoning, the ruptured packets were encountered in the upper gastrointestinal tract and several other partially ruptured packets were also found, implying that poor packaging was the cause of rupture. Obstructing packets should be removed but non-obstructing, unruptured packets encountered in the colon may safely be allowed to pass spontaneously. All cases of packet rupture with cocaine toxicity should have immediate surgery. PMID- 15892389 TI - Retrospective review of leptospirosis in Guadeloupe, French West Indies 1994 2001. AB - Demographic, clinical, biological and personal data were obtained from patients hospitalized with symptoms of leptospirosis in the Hospital of Pointe a Pitre, Guadeloupe, French West Indies from 1994 to 2001. Of the 897 screened patients, 212 were acute cases, 607 were non-infected and 78 were undetermined cases. There was no predominant age group. Leptospirosis transmission followed the rainfall cycle and was greater in rural areas. Jaundice and conjunctival suffusion were significantly more frequent in cases than non-cases. Males, professions considered to be at risk and contact with swine or bovine were associated with infection. Serogroups Icterohaemorrhagiae, Cynopteri, Australis, Sejroe, Pomona and Ballum were serovars presumed responsible for acute cases. PMID- 15892390 TI - Oesophageal foreign bodies at the University Hospital of the West Indies. AB - A retrospective study was conducted of 97 patients with oesophageal foreign bodies (EFB). The patients were admitted to assess characteristics of EFB, modes of presentation and radiological and endoscopic findings. The patients were from the University Hospital of the West Indies and most (42%) were over the fourth decade of life. The commonest EFB were bones. A negative radiological finding was not a reliable means to select patients for endoscopy. Oesophagoscopy is a reliable method in the treatment of EFB impaction. There were no major complications or deaths. PMID- 15892391 TI - Critical incidents contributing to the initiation of substance use and abuse among women attending drug rehabilitation centres in Trinidad and Tobago. AB - The aim of this study was to determine the critical incidents that contribute to the initiation of substance use and abuse among women in Trinidad and Tobago. Twenty women were randomly selected from 46 women currently attending 43 drug rehabilitation centres, Narcotics Anonymous and Alcoholics Anonymous groups in Trinidad and Tobago. In-depth semi-structured interviews using the critical incident technique were conducted. Interviews were recorded, transcribed and analyzed. Concepts, categories and themes were determined by team study and group discussion. The critical incidents that influenced women to initiate the use and abuse of substances fell into eight major themes: factors intrinsic to the individual woman, family factors, social and environmental factors, life stresses, relationship issues, abuse, peer pressure and substance use and abuse as a coping mechanism. The results imply that the factors contributing to the initiation of substance use and abuse among women in Trinidad and Tobago are many and complex. As such any attempt to address this issue requires a broad-based approach. Such an approach should address family use of such substances, societal acceptance of them, availability, the self-esteem of the individual woman and her ability to cope with peer and internal stresses. PMID- 15892392 TI - Vomiting post tonsillectomy at the University Hospital of the West Indies. AB - A three-year observational study of patients undergoing tonsillectomy at the University Hospital of the West Indies was conducted to determine the incidence of postoperative vomiting. Data were collected to assess possible risk factors for vomiting as well as possible alleviating agents. Two hundred and fifty-two patients were included in the study and a thirteen per cent incidence of postoperative vomiting was found. This is significantly less than that quoted in other studies (40-73%). Results also showed that steroids significantly reduced the incidence of postoperative vomiting in the study population. Muscle relaxants reversal agents and antibiotics particularly co-trimoxazole and ceftriaxone significantly increased its incidence. Usual antiemetic agents including dimenhydrinate (gravol) and promethazine (phenergan), as well as drugs known to possess antiemetic properties such as midazolam and propofol, lacked any significant protective effect against emesis. Opioid analgesia, inhalational induction and blood loss of greater than 10% of estimated blood volume appeared to increase emesis but failed to achieve statistical significance. PMID- 15892393 TI - Use what you have--biological assistance for the treatment of heart failure in the Caribbean. AB - Cardiovascular disease represents the main cause of death among adults in the Caribbean. Primary and secondary care facilities are efficiently managed. Cardiac surgical and interventional facilities, however, exist only in a small number of territories and are mainly privately funded and are only accessible to few patients. Patients with end-stage heart failure (ESHF) are given few options apart from palliative care or to seek treatment outside of the region. Transplantation remains the 'gold standard' therapy for ESHF. Establishing a Caribbean cardiac transplantation programme would require legislative and infrastructure changes. Tissue rejection poses a problem and expensive immunosuppressants are needed. Mechanical assist devices are costly and associated with complications such as haemorrhage, thrombosis and infections. Both forms of therapy require significant technical and financial investment and do not appear to be economically viable for the Caribbean. The use of the patient's own skeletal muscle to perform biological cardiac assistance is potentially the ideal alternative. The skeletal muscle is conditioned by electrical stimulation to become fatigue resistant. It is then transposed and harnessed as an auxilliary circulatory pump. The required muscle stimulators are relatively inexpensive and the surgical techniques and postoperative care are not overly demanding. We discuss the financial and research implications of treating patients from the Caribbean who have end-stage heart failure. PMID- 15892394 TI - High risk health behaviours among adult Jamaicans. AB - The purpose of this study was to assess the prevalence of high risk health behaviours among adult Jamaicans aged 15-49 years in 2000, and to compare the results with the 1993 survey. A nationally representative sample of 2013 persons aged 15-74 years was surveyed in 2000 using cluster sampling in the Jamaica Healthy Lifestyle Survey (Wilks et al, unpublished). Interviewer administered questionnaires and anthropometrical measurements were done. Data for a sub-sample of adults aged 15-49 years were analyzed The sub-sample included 1401 persons (473 men and 928 women). Significantly more men (18.6%) than women (4.3%) reported never having had a blood pressure check (p = 0.0001). Approximately one third of the women reported that they had never had a Pap smear (36.0%) or a breast examination (31.2%). Current cigarette smoking was reported in 28.6% of men and 7.7% of women (OR 3.73 CI 2.71, 5.15), while 49.0% of men and 15.0% of women ever smoked marijuana (OR 3.28 CI 2.56, 4.20). Significantly more men (28.0%) than women (11.7%) reported ever having a sexually transmitted disease (OR 2.93 CI 2.16, 3.97); having more than one sexual partner in the past year (49.1% vs 11.4%, OR 4.31 CI 3.22, 5.76) and usually using a condom during sexual intercourse (55.3% vs 40.5%, OR 1.3 CI 1.11, 1.68). Between 1993 and 2000, significant trends include: more persons reported having a blood pressure check, a reduction in multiple sexual partners, increased condom use at last sex (women), reduced crack/cocaine use (males) and increased marijuana smoking. Although there were some significant positive lifestyle trends between 1993 and 2000, high risk behaviours remain common among Jamaican adults. Comprehensive health promotion programmes are needed to address these risk behaviours. PMID- 15892395 TI - Juggling career aspirations with quality and relevance to the Caribbean. PMID- 15892396 TI - Popliteal artery transection associated with blunt knee injury: controversies in management--a case report. PMID- 15892397 TI - Chronic cholelithiasis with gallstones lodged in an isolated subserosal intramural gastric pouch. PMID- 15892398 TI - Pituitary apoplexy associated with ptosis. AB - Pituitary tumours are the most common sellar masses, frequently presenting with visual impairment and endocrine abnormalities. Two cases of pituitary tumour presenting with ptosis are reported. PMID- 15892399 TI - A case of referred pain following a pedicled medial gastrocnemius muscle flap. PMID- 15892400 TI - Children's rights in Europe. PMID- 15892401 TI - Effects of epoetin alfa on blood transfusions and postoperative recovery in orthopaedic surgery: the European Epoetin Alfa Surgery Trial (EEST). AB - BACKGROUND AND OBJECTIVE: Preoperative epoetin alfa administration decreases transfusion requirements and may reduce transfusion complications, such as postoperative infection due to immune suppression and thus hospitalization time. This study examined the impact of preoperative epoetin alfa administration on postoperative recovery and infection rate. METHODS: In an open randomized controlled multicentre trial in patients undergoing orthopaedic surgery, the effects of preoperative administration of epoetin alfa vs. routine care were compared in six countries. Haemoglobin (Hb) values, transfusions, time to ambulation, time to discharge, infections and safety were evaluated in patients with preoperative Hb concentrations 10-13g dL(-1) (on-treatment population: epoetin n = 460; control n = 235), from study entry until 4-6 weeks after surgery. Outcome was also compared in patients with and without transfusion. RESULTS: Epoetin-treated patients had higher Hb values from the day of surgery until discharge (P < 0.001) and lower transfusion rates (12% vs. 46%; P < 0.001). Epoetin treatment delivered no significant effect on postoperative recovery (time to ambulation, time to discharge and infection rate). However, the time to ambulation (3.8+/-4.0 vs. 3.1+/-2.2days; P < 0.001)and the time to discharge (12.9+/-6.4 vs. 10.2+/-5.0 days; P < 0.001) was longer in the transfused than in the non-transfused patients. Side-effects in both groups were comparable. CONCLUSIONS: Epoetin alfa increases perioperative Hb concentration in mild-to moderately anaemic patients and thus reduces transfusion requirements. Patients receiving blood transfusions require a longer hospitalization than non-transfused patients. PMID- 15892402 TI - Anaesthesia and circulating blood volume. AB - BACKGROUND AND OBJECTIVE: The exact change in circulating blood volume (BV) during general anaesthesia is still unknown because there is no standard method of evaluating BV. We evaluated the changes in BV by general anaesthesia using simple and easy estimation methods. METHODS: Fourteen patients scheduled for minor surgery under general anaesthesia were enrolled. Propofol and vecuronium bromide were used for the induction of anaesthesia, and anaesthesia was maintained with sevoflurane and nitrous oxide. Haematocrit (Hct), total protein concentration (TP), as well as colloid osmotic pressure (COP) measured using a colloid osmometer, were determined before anaesthesia, and 30, 60 and 90 min after the induction of general anaesthesia. BV was calculated using Allen's formula and the changes in Hct, TP and COP. The estimated BV was compared with directly measured BV using indocyanine green dilution method (BV(ICG)). RESULTS: Hct, TP and COP significantly decreased after the induction of anaesthesia (Hct: 42.1-39.4%; TP: 7.3-6.9 g dL(-1); COP: 23-19 mmHg). The calculated BV as well as BV(ICG) significantly increased after induction of anaesthesia (calculated by COP: 4.13-5.03 L; BV(ICG): 4.54-5.56 L). The change rate in BV calculated by the change of COP was larger than other calculated BVs, and was approximated to the change rate in BV(ICG). After emergence from anaesthesia, all values tended to return to baseline. CONCLUSIONS: General anaesthesia increases BV. The value of BV calculated from the change in COP was most changeable. PMID- 15892403 TI - Comparative study of intubating conditions at the first minute with suxamethonium, rocuronium and different priming techniques of rocuronium. AB - BACKGROUND AND OBJECTIVE: To evaluate orotracheal intubation conditions after 1 min. PATIENTS AND METHODS: A prospective randomized study with 376 adult American Society of Anesthesiologists (ASA) Grade I-III patients. Each patient received propofol, fentanyl and either suxamethonium (1 mg kg(-1)) or rocuronium. The intubating dose of rocuronium (2 x ED95) was preceded 4 min earlier by saline, or a 0.1 x ED95 priming dose of rocuronium, atracurium, cis-atracurium, vecuronium or mivacurium. Intubating conditions were graded as excellent, good or poor with respect to laryngoscopy, vocal cord position and movement and reaction to intubation and/or cuff inflation. RESULTS: There were significant differences (P < 0.05) in laryngoscopy between suxamethonium and rocuronium primed with saline, atracurium or cis-atracurium. With respect to vocal cord position and movement during intubation, rocuronium without priming differed significantly from all other groups and for reaction to insertion of tracheal tube and/or cuff inflation. Rocuronium without priming differed significantly from all other groups except for rocuronium primed with itself. The mivacurium group showed more signs of pre-curarization than other groups (P < 0.05). There were significant differences between rocuronium alone and the other groups when final intubating conditions were compared. CONCLUSIONS: Priming rocuronium with 0.1 x ED95 of vecuronium, rocuronium, atracurium or cis-atracurium is a safe technique and did not increase risk of pre-curarization in healthy patients. PMID- 15892404 TI - Plasma concentrations of nitric oxide products and cognitive dysfunction following coronary artery bypass surgery. AB - BACKGROUND AND OBJECTIVE: Prospective longitudinal studies now indicate that cognitive dysfunction following coronary artery bypass surgery (CABG) is both common and persistent. This dysfunction is due in part to the inflammatory response and cerebral ischaemia-reperfusion, with nitric oxide (NO) as an important mediator of both. We hypothesized that a clinically significant association exists between plasma concentrations of nitrate/nitrite (NO3-/NO2-) and cognitive dysfunction after CABG. METHODS: Cognitive assessment was performed on 36 adult patients the day before CABG, on the fourth postoperative day and 3 months postoperatively. Patient spouses (n = 10) were also studied. RESULTS: A new cognitive deficit was present in 22/36 (62%) 4 days postoperatively and in 16/35 (49%) of patients, 3 months postoperatively. Patients who had cognitive dysfunction 3 months postoperatively were more likely to have cognitive dysfunction and increased plasma NO3-/NO2- concentrations compared to the non deficit group preoperatively (22.6 (9.2) vs. 27.6 (8.4)) (P = 0.002). Plasma NOx (NO3- plus NO2-) concentrations were greater in patients with cognitive dysfunction 3 months postoperatively, 2 h (24.2 (6.3) vs. 19.1 (5.2)) (P = 0.002), and 12 h postoperatively (24.8 (7.6) vs. 18.8 (5.6)) (P = 0.001). There was, however, a time course similarity in NOx elevations for both deficit and non deficit groups. CONCLUSIONS: Perioperative plasma NOx concentrations do not serve as an effective biomarker of cognitive deficit after CABG. PMID- 15892405 TI - Effect of aprotinin on in vitro cerebral endothelial ICAM-1 expression induced by astrocyte-conditioned medium. AB - BACKGROUND AND OBJECTIVE: Aprotinin administration may decrease the incidence of stroke associated with coronary artery bypass surgery by an unknown mechanism. Astrocytes exposed to hypoxia produce proinflammatory cytokines and upregulate intercellular adhesion molecule (ICAM)-1 on cerebral endothelium. This study investigated the effects of aprotinin on cerebral endothelial activation by hypoxic astrocytes in vitro. METHODS: Mouse astrocytes were exposed to hypoxia in an anaerobic chamber for 4 h followed by reoxygenation for 24 h. Astrocyte conditioned medium (ACM) collected from mouse astrocytes subjected to hypoxia/reoxygenation (HR) or normoxia were applied to mouse cerebral endothelial cell (MCEC) cultures for 4 and 24 h in normoxia. Endothelial cells were preincubated for 1 h with aprotinin (1600 KIU mL(-1)) prior to exposure to ACM. Flow cytometry was used to estimate endothelial ICAM-1 expression. Interleukin (IL)-1beta space concentrations in ACM were estimated with enzyme-linked immunosorbent assay (ELISA). Repeated comparisons were made using analysis of variance (ANOVA) and post hoc Tukey test as appropriate. P < 0.05 was considered significant. Data is presented as mean (standard deviation, SD). RESULTS: MCEC ICAM-1 expression was greater after 24 h exposure to HR-ACM compared to normoxic ACM (mean channel flouresence (MCF) 107.5 (4.5) vs. 74.3 (4.5), respectively, P < 0.001). ICAM-1 expression was decreased by aprotinin preincubation compared to control (MCF 91.0 (1.1) vs. 107.5 (2.1), P = 0.006). Supernatant IL-1beta concentrations in astrocytes exposed to HR were greater than those exposed to normoxia (7.1 (0.2) vs. 4.1 (0.2), P = 0.01). CONCLUSIONS: This may be a neuroprotective mechanism associated with aprotinin administration. PMID- 15892406 TI - Phosphodiesterase-III-inhibition with amrinone leads to contracture development in skeletal muscle preparations of malignant hyperthermia susceptible swine. AB - BACKGROUND AND OBJECTIVE: The phosphodiesterase-III (PDE-III) inhibitor enoximone induced marked contractures in skeletal muscle specimens of malignant hyperthermia (MH) susceptible (MHS) human beings and swine. Whether this is a substance specific effect of enoximone or caused by inhibition of PDE-III remained unclear. Therefore, the effects of the PDE-III inhibitor amrinone in porcine MH normal (MHN) and MHS skeletal muscles were investigated. METHODS: MH trigger-free general anaesthesia was performed in eight MHS and eight MHN swine. The MH status of the swine was determined by detection of the Arg615-Cys point mutation on chromosome 6 indicating MH susceptibility. Skeletal muscle specimens were excised for the in vitro contracture tests with amrinone. Amrinone was added cumulatively every 5 min to muscle specimens in order to obtain organ bath concentrations between 20 and 400 micromol L(-1). The in vitro effects of amrinone on muscle contractures and twitches were measured. RESULTS: Amrinone induced contractures in all skeletal muscle preparations. MHS muscles developed contractures at significantly lower bath concentrations of amrinone than MHN muscles. Contractures of MHS compared to MHN muscles were significantly larger at bath concentrations of 80, 100, 150, 200 and 400 micromol L(-1) amrinone. Muscle twitches remained unchanged up to and including 200 micromol L(-1) amrinone. CONCLUSIONS: Inhibition of PDE-III in general elicited higher contractures in MHS than in MHN muscles. Therefore, a contribution of PDE-III and the cyclic adenosine monophosphate (cAMP) system in the pathophysiology of MH must be suspected. PMID- 15892407 TI - Pseudocholinesterase activity increases and heart rate variability decreases with preoperative anxiety. AB - BACKGROUND AND OBJECTIVE: The objective of this study was to determine the influence of preoperative anxiety on the activity of plasma cholinesterase and heart rate (HR) variability. METHODS: A total of 50 subjects were studied, 25 male patients one day preoperatively and 25 male volunteers without surgical intervention as a control group. Blood samples were taken to determine plasma cholinesterase activity. HR variability was recorded for a period of 256 beat-to beat intervals and analysed by frequency domain analysis into very low frequency (VLF: 0.02-0.04 Hz), low frequency (LF: 0.04-0.15 Hz) and high frequency (HF: 0.15-0.4 Hz). LF/HF ratio and total power over the 0.02-0.4 Hz range were calculated. Anxiety levels were assessed using the hospital anxiety and depression scale, the self-rating anxiety scale and a visual analogue scale. RESULTS: The patient group had significantly higher anxiety scores. Plasma cholinesterase activity was significantly higher in patients vs. controls (6646 vs. 5324 units L(-1)). Total power, LF and HF were significantly lower in the patients (1489 vs. 2581 ms2; 656 vs. 1186 ms2; 491 vs. 964 ms2, respectively). CONCLUSIONS: Preoperative anxiety increases plasma cholinesterase activity and decreases HR variability. PMID- 15892408 TI - Monitoring of selective antegrade cerebral perfusion using near infrared spectroscopy in neonatal aortic arch surgery. AB - BACKGROUND AND OBJECTIVE: To prevent neurological complications, low-flow antegrade cerebral perfusion (ACP) is used during repair of complex congenital heart defects. To overcome technical problems, continuous monitoring of cerebral blood flow and oxygenation is mandatory. The aim of the study was to evaluate the effect of different ACP flow rates on cerebral oxygen saturation obtained by near infrared spectroscopy. METHODS: Ten consecutive neonates undergoing Norwood stage I were included. In addition to near infrared spectroscopy (Invos 5100; Somanetics Corp., USA) on both hemispheres, mean arterial pressure and transcranial Doppler flow velocity were measured continuously and arterial and jugular venous oxygen saturation intermittently. Cerebral oxygen extraction ratio was calculated. Measurement points were obtained after starting bypass, during ACP with flow rates of 30, 20 and 10 mL kg(-1) min(-1) and immediately after ACP. ANOVA and Tukey-Kramer multiple comparison test were used for statistics. RESULTS: The near infrared spectroscopy signal could be obtained in all children at all measurement points, whereas transcranial Doppler failed in 1 neonate at a flow rate of 30 mL kg(-1)min(-1), in 3 neonates at 20 mL kg(-1) min(-1) and in 4 neonates at 10 mL kg(-1)min(-1). With the reduction of flow there was a significant decrease of cerebral oxygen saturation on both hemispheres (right: 78+/-8 to 72+/-9 and 66+/-8, P < 0.001; left: 71+/-7 to 65+/-7 and 60+/-7, P < 0.001), of jugular venous oxygen saturation (94+/-6 to 89+/-13 and 83+/-15, P < 0.001) and a significant increase in oxygen extraction ratio (9.1+/-8 to 14.8+/ 14 and 21+/-16, P < 0.001) respectively, for 30, 20, 10 mL kg(-1)min(-1). CONCLUSION: Near infrared spectroscopy reliably detects flow alterations during ACP with profound hypothermia. PMID- 15892409 TI - A combination of lidocaine and nitrous oxide in oxygen is more effective in preventing pain on propofol injection than either treatment alone. AB - BACKGROUND AND OBJECTIVE: Propofol is an intravenous (i.v.) anaesthetic agent that possesses many of the qualities of an ideal anaesthetic agent. The most significant side-effect associated with propofol is pain on injection. Despite optimal therapy, the incidence of pain on propofol injection remains a problem. This prospective, randomized, double blinded study evaluated the effect of three different treatment strategies in decreasing pain on propofol injection. METHODS: We studied 102 adult, ASA I-II patients, scheduled for elective surgical procedures. Combination of i.v. lidocaine and nitrous oxide (N2O) in oxygen (O2) inhalation pre-treatment was compared with either treatment alone in the prevention of pain on propofol injection. A standard propofol injection technique and scoring system, to measure the pain on injection was used. RESULTS: Demographic variables were similar between the groups. The incidence of no pain on propofol injection was similar in the lidocaine and N2O groups (63.6% vs. 57.5%) (95% confidence interval (CI): 0.17-0.29, P = 0.61). Combination therapy was associated with a greater incidence of no pain on injection (84% vs. 63.6%) (95% CI: 0.06-0.48, P = 0.04). CONCLUSION: Combination of i.v. lidocaine and N2O in O2 inhalation pre-treatment is more effective than either treatment alone in decreasing pain on propofol injection. PMID- 15892410 TI - Time course and train-of-four fade of mivacurium block during sevoflurane and intravenous anaesthesia. AB - BACKGROUND AND OBJECTIVE: Volatile anaesthetics inhibit nicotinic acetylcholine receptors at clinically relevant concentrations with higher affinity for the neuronal nicotinic receptor. The inhibitory effects of propofol on nicotinic receptors have only been documented at supraclinical concentrations. The aim of this study was to determine recovery properties and train-of-four (TOF) fade of mivacurium during sevoflurane and propofol anaesthesia, in order to examine any differences both in the enhancement of the neuromuscular block (postjunctional effects) and in TOF fade (prejunctional effects). METHODS: Twenty ASA I-II adult patients were randomly allocated to maintenance of anaesthesia with sevoflurane (end-tidal concentration 2%) or propofol. Neuromuscular block was assessed by acceleromyography and a single dose of mivacurium (0.15 mg kg(-1)) was administered (in the sevoflurane group after 30 min of exposure to sevoflurane). We measured time for recovery of the first twitch of the TOF (T1) from 25-75%, time from 25% recovery of T1 to achieving a TOF ratio (TOFR) of 0.8, TOFR at 50%, 75% and 90% recovery of T1, and height of T1 at TOFR of 0.7 and 0.9. Data were tested using t-test for independent samples. RESULTS: Recovery times (mean (95% confidence interval, CI)) of mivacurium in the sevoflurane group (T1 25-75%, 11.3 (8.1-14.5) min; T1 25%-TOFR0.8, 19.1 (15.7-22.5) min) were significantly longer (P < 0.05) than in the propofol group (T1 25-75%, 6.5 (5.2-7.7) min; T1 25% TOFR0.8, 11.3 (7.8-10.3) min). No differences were found in the relations between TOFR and T1 or vice versa, between the groups. CONCLUSIONS: Recovery times after a single dose of mivacurium were prolonged by sevoflurane compared with propofol but no differences in TOF fade were observed between the two anaesthetics. PMID- 15892411 TI - Complaints of sore throat after tracheal intubation: a prospective evaluation. AB - BACKGROUND AND OBJECTIVE: Sore throat and hoarseness rank, besides pain and nausea, among the most frequent subjective complaints after tracheal intubation for general anaesthesia. Our intention was to determine the incidence of postoperative sore throat from a large sample of patients and thus to identify the most important associated factors. METHODS: We prospectively followed up 809 adult patients who underwent elective surgical interventions and examined their history, the applied anaesthetic techniques, perioperative course and the occurrence, intensity and duration of postoperative throat complaints. The assignment and professional experience of the involved intubators were also assessed. The influence of a multitude of variables on postoperative throat complaints was statistically analysed. RESULTS: Postoperative sore throat was present in 40% overall being significantly higher in female than in male (44% vs. 33%; P = 0.001). The mean pain intensity in the affected patients (n = 323) was 28+/-12 mm on a visual analogue scale where 0 = no pain and 100 = extreme pain. The average duration was 16+/-11 h. Main factors associated with throat complaints were female sex; history of smoking or lung disease, duration of anaesthesia, postoperative nausea, bloodstain on the endotracheal tube and natural teeth. We could find no influence on the occurrence or intensity of throat complaints by the professional assignment or the length of professional experience of the personnel involved. CONCLUSIONS: Postoperative throat complaints frequently arise after tracheal intubation for general anaesthesia in the first 2 postoperative days, but they are of limited intensity and duration. PMID- 15892412 TI - Additional work of breathing and breathing patterns in spontaneously breathing patients during pressure support ventilation, automatic tube compensation and amplified spontaneous pattern breathing. PMID- 15892413 TI - Fentanyl reduces cortisol and blood glucose changes during cataract surgery under retrobulbar anaesthesia. PMID- 15892414 TI - Anaesthesia in vitreo-retinal surgery. PMID- 15892415 TI - The use of transillumination for peripheral venous access in paediatric anaesthesia. PMID- 15892416 TI - Severe burns during magnetic resonance examination. PMID- 15892417 TI - Over-interpretation of the TEI-index. PMID- 15892418 TI - Gastric volume and pulmonary aspiration. PMID- 15892419 TI - A case of normal ropivacaine concentration causing grand mal seizure after epidural injection. PMID- 15892420 TI - Estimating clinically significant differences in quality of life outcomes. AB - OBJECTIVE: This report extracts important considerations for determining and applying clinically significant differences in quality of life (QOL) measures from six published articles written by 30 international experts, in the field of QOL assessment and evaluation. The original six articles were presented at the Symposium on Clinical Significance of Quality of Life Measures in Cancer Patients at the Mayo Clinic in April 2002 and subsequently were published in Mayo Clinic Proceedings. PRINCIPAL FINDINGS: Specific examples and formulas are given for anchor-based methods, as well as distribution-based methods that correspond to known or relevant anchors to determine important differences in QOL measures. Important prerequisites for clinical significance associated with instrument selection, responsiveness, and the reporting of QOL trial results are provided. We also discuss estimating the number needed to treat (NNT) relative to clinically significant thresholds. Finally, we provide a rationale for applying group-derived standards to individual assessments. CONCLUSIONS: While no single method for determining clinical significance is unilaterally endorsed, the investigation and full reporting of multiple methods for establishing clinically significant change levels for a QOL measure, and greater direct involvement of clinicians in clinical significance studies are strongly encouraged. PMID- 15892421 TI - The contextual model of HRQoL: a paradigm for expanding the HRQoL framework. AB - Cancer is the second leading cause of death in most US populations. Unfortunately, ethnic minority status is associated with increased later stage at diagnosis, greater incidence for many cancer sites, differential treatments, greater mortality and morbidity. The government and public health focus on health disparities, evident in several documents including Healthy People 2010, Unequal Treatment and the Nation's Investment in Cancer Research, are spurring interest in research with ethnic minority populations. Research investigating the health related quality of life (HRQoL) among ethnic minority cancer survivors is new and growing. However, there is a dearth of research that addresses theoretical frameworks in cross cultural research. In conducting research with diverse populations, appropriate theoretical grounding that is responsive to cultural and socioecological contexts must be considered. This paper will discuss the contextual model of HRQoL, a comprehensive framework developed to expand the traditional HRQoL framework to facilitate culturally and socioecologically responsive research. This model may provide a more comprehensive theoretical framework to investigate certain areas of health disparities and risk factors for poor outcomes in HRQoL research with cancer survivors. PMID- 15892422 TI - Literature review of instruments to assess health-related quality of life during and after menopause. AB - BACKGROUND AND OBJECTIVES: Menopause is a physiological event occurring in women at about the age of 50. It signals the end of the reproductive years and is associated with signs of estrogen deficiency having a considerable impact on women's health-related quality of life (HRQoL). The most common form of treatment is hormone replacement therapy (HRT). Studies have shown negative events can arise from long-term use of HRT. The aim of this review is to determine if there are any HRQoL instruments that address the impact of menopausal symptoms including positive and negative effects of HRT. METHODS: The following eight instruments were identified: Greene Climacteric Scale, Women's Health Questionnaire (WHQ), Qualifemme, Menopause-Specific QOL Questionnaire (MENQOL), Menopausal Symptoms List (MSL), Menopause Rating Scale (MRS), Menopausal Quality of Life Scale (MQOL), and the Utian Quality of Life Scale (UQOL). RESULTS: All instruments reviewed proved to be reasonably structured and have their place in applied research. None were found that addressed all aspects of the impact of HRT on HRQoL. CONCLUSION: In order to capture the possible short-term side effects of HRT on HRQoL, it is necessary to modify one or more of the existing instruments or develop a new instrument applicable in many different countries and languages. PMID- 15892423 TI - The quality of life impact of acute exacerbations of chronic bronchitis (AECB): a literature review. AB - BACKGROUND: The impact of acute exacerbations of chronic bronchitis (AECB), a common consequence of chronic obstructive pulmonary disease (COPD), is extensive, with symptoms ranging from mild to life threatening. Health-related quality of life (HRQL) is impaired in patients with COPD, but little is known about the direct effect of exacerbations on HRQL. METHODS: MEDLINE and EMBASE literature searches were conducted; reference lists of identified articles were reviewed. RESULTS: Eighteen studies reporting the impact on HRQL of acute exacerbations were identified. Study design and patient population varied. Six studies evaluated HRQL once; only four studies used both generic and disease-specific HRQL measures. Cross-sectional studies reported HRQL decrements during exacerbations and suggested that HRQL is a good predictor of health care resource utilization. Pharmacological treatment led to within-group improvements following AECBs. Non-pharmacological intervention studies were small and inconclusive. Longitudinal studies, assessing pharmacological and non-pharmacological interventions, found that HRQL improved from exacerbation to recovery, with responsiveness depending on sensitivity of the measure. Frequency of exacerbations was a significant predictor of HRQL. CONCLUSIONS: Exacerbations lead to substantial reductions in HRQL, both in physical as well as other domains. Further research should assess the impact of specific treatment regimens and the timeline for the recovery process. PMID- 15892424 TI - Life values in patients with COPD: relations with pulmonary functioning and health related quality of life. AB - Theories of coping and response shift have suggested that emotional adaptation is related to value changes, e.g. a deemphasized importance of lost life values and an enlargement of the scope of values. Perceived attainment and importance of 82 life values were examined in 65 patients with chronic obstructive pulmonary disease (COPD) and related to clinical and Health-Related Quality of Life (HRQL) measures. The life values covered 10 dimensions--harmony, positive relations, involvement, mobility, communication, knowledge, responsibility, comfort, religion and health. Forty-six of the patients were followed up after 1 year. The patients with COPD were compared with a healthy control group, a group of neurologically impaired and a non-disabled group representing the general population. Significant congruence was found between importance and attainment ratings in all groups (correlations from 0.44 to 0.53), suggesting that both impaired and healthy persons tend to perceive that they have what they find important in life. Congruence was significantly related to mood (correlations from 0.28 to 0.40), but not to functional status or clinical data. Compared to the healthy responders, the patients with COPD had significantly lower attainment ratings in health, mobility, involvement, but no differences were found for importance ratings. No evidence was found that they had replaced unattainable values with new available values, and no changes over time of perceived values were found. This suggests that patients with COPD do not seem to adapt by means of changing their value orientation. PMID- 15892425 TI - Influence of exercise activity on quality of life in long-term breast cancer survivors. AB - BACKGROUND: Behavioral and lifestyle factors may influence quality of life (QOL) outcomes in breast cancer survivors. METHODS: Information on QOL (Short Form-36, SF-36), lifestyle and survivorship was collected during telephone interviews with 374 breast cancer patients, diagnosed between 1983 and 1988 at ages 40 years or younger and interviewed, on average 13.2 years following diagnosis. These women previously participated in a case-control study soon after their diagnoses, providing information on breast cancer risk factors including exercise activity. We examined the impact of changes in exercise activity (comparing pre- to post diagnosis levels) on the SF-36 mental and physical health summary scales using regression analyses. RESULTS: A positive change in exercise activity was associated with a higher score on the SF-36 physical health summary scale at follow-up (p = 0.005). Change in exercise activity was not associated with the SF 36 mental health summary scale score. Patients who increased their activity levels did not differ from those who did not in terms of medical or demographic characteristics. CONCLUSION: This study provides one of the longest follow-up periods of breast cancer survivors to date among studies that focus on QOL and is unique in its focus on women diagnosed at a young age. Our results confirm high levels of functioning and well-being among long-term survivors and indicate that women whose exercise activity increased following diagnosis score higher on the SF-36 physical health summary scale. These findings suggest a potential role for exercise activity in maintaining well-being after a cancer diagnosis. PMID- 15892426 TI - Development and evaluation of an instrument to assess social difficulties in routine oncology practice. AB - PURPOSE: To develop and preliminarily evaluate a Social Difficulties Inventory (SDI) for use in oncology practice. METHODS: Item generation from patients (n = 96), staff (n = 49) and the literature. Questions constructed and pre-tested (n = 42) resulting in a 22-item questionnaire. Psychometric evaluation (n = 271) assessed frequency of endorsement, factor structure, summated scales and construct validity followed by randomisation to criterion validity or test-retest arms. RESULTS: Items met preset frequency of endorsement criteria. Factor analysis revealed a four-factor structure, three clearly definable, (1) Physical ability, (2) Providing for the family and (3) Contact with others, explaining 45.8% variance. Summated scales developed from these demonstrated good reliability (Cronbach's alpha > 0.7) and were used to test and confirmed construct validity. Several comparators from the Life Events and Difficulties Schedule (LEDS) and Cancer Rehabilitation Evaluation System Short Form (CARES-SF) tested criterion validity and majority agreements were moderate to good. Test retest reliability was good with most kappa values > 0.6. One item was eliminated resulting in a 21-item questionnaire. CONCLUSION: The SDI is relevant and easy to understand. Initial psychometric evaluation was encouraging. Ongoing work to evaluate the clinical meaning and utility of the instrument and to examine the relationships between SDI scores and clinical outcomes will provide guidance about its usefulness as an assessment tool in routine oncology practice. PMID- 15892427 TI - A prospective comparison of quality of life measures for patients with esophageal cancer. AB - Among the most widely used instruments to assess quality of life (QOL) in patients with cancer are the European Organization for Research and Treatment of Cancer core questionnaire (EORTC QLQ-C30) and the Functional Assessment of Chronic Illness Therapy, cancer instrument (FACT-G). This study compared these approaches in patients who had undergone esophagectomy for cancer. The EORTC core questionnaire and esophageal module and the FACT-G and esophageal scale were completed by 57 patients. Missing data, relationships between QOL scales and analyses of patients' preferences were examined. There were 14/2736 (0.5%) missing items from EORTC questionnaires and 45/2565 (1.8%) from FACT instruments (p < 0.01). Relationships between corresponding generic EORTC and FACT scales were average to good (r > 0.57) except for the social function scale (r = 0.01). EORTC symptom scores were moderately correlated with the FACT general scale, but poorly related to the FACT esophageal scale (r < 0.28). EORTC swallowing scores were moderately correlated with all FACT scales. The FACT-E and EORTC QLQ-C30 measure assess similar generic aspects of QOL (except social function). EORTC esophageal symptom scores relate poorly to FACT esophageal scales, except for swallowing. Choice of QOL measure after esophagectomy for cancer depends upon outcomes of interest. Future studies will determine which instruments are appropriate in each context. PMID- 15892428 TI - A two-year prospective study of the health-related quality of life of children with chronic illness--the parents' perspective. AB - The aim of this study was to assess prospectively changes in the health-related quality of life (HRQL) of children and adolescents with diabetes, asthma or cystic fibrosis (CF). One hundred and twenty-two parents of children aged 10-16 years with asthma, diabetes, or CF were recruited from specialist paediatric clinics. Parents described their children's HRQL using the Child Health Questionnaire (PF98) at baseline, 6, 12, 18 and 24 months post-baseline. They reported that the general health of children with CF was significantly worse than that of children with asthma and diabetes at baseline. In other domains there were few differences between the HRQL of children in the three groups. In several domains, the HRQL of children with asthma or diabetes improved over the 2 years of the study. This improvement was less evident for children with CF. PMID- 15892429 TI - Is treatment adherence associated with better quality of life in children with sickle cell disease? AB - The association of treatment adherence with quality of life (QOL) and the role of sickle cell disease complications were explored in children with sickle cell disease. Primary caregivers of 43 children, ages 5 years and older, and 21 children, ages 8 years and older, completed a standardized measure of QOL during an admission for pain or fever to the hematology acute care unit. Adherence was measured through medical staff ratings, caregiver-report of sickle cell disease related care activities, and matching of medical staff standard recommendations for treatment of pain and fever with sickle cell disease-related care activities. Sickle cell disease complications were assessed via medical file review. Pearson correlation coefficients indicated that better adherence was associated with poorer overall QOL. In follow-up analyses, although sickle cell disease complications were associated with adherence, it did not explain the negative correlation of adherence with QOL. Higher treatment adherence may interfere with activities that contribute to QOL for some children. Further research to investigate the role of sickle cell disease complications, as well as psychosocial factors, in determining both treatment adherence and QOL is suggested. PMID- 15892430 TI - Self-perceived quality of life of children and adolescents with physical disabilities in Hong Kong. AB - Although illnesses and diseases are thought to adversely affect quality of life (QoL), whether children who have physical disabilities (PD) from a young age adapt to the effect of developmental disabilities has rarely been investigated. This study attempted to assess the subjective wellbeing, and examine the correlation between objective and subjective QoL, of children with PD. Using a self-reported non-disease-specific questionnaire, the QoL of 72 young persons (13.5+/-2.0 years) with PD was contrasted with those who do not have disabilities (n = 510; age-matched). MANOVA analyses revealed that the PD group had lower objective QoL score (63.0+/-7.4 vs. 66.8+/-5.7, p < 0.001) but the two groups were not significantly different in subjective QoL score (70.9+/-11.4 vs. 69.6+/ 13.6, p = 0.466). No correlation was found between objective and subjective QoL in the PD group (r ranged from 0.06 to 0.19), while weak to medium correlations (r ranged from 0.03 to 0.41) were observed for the controls. The apparent detachment of subjective feeling and objective circumstances in the PD group may reflect adjustment to developmental disabilities. PMID- 15892431 TI - Quality of life in multiple sclerosis patients with urinary disorders: discriminative validation of the English version of Qualiveen. AB - The Qualiveen questionnaire is a urinary disorder (UD)-specific health related quality of life (HRQL) instrument. Recent data suggests Qualiveen has excellent validity in French-speaking multiple sclerosis (MS) patients. AIM: To assess discriminative measurement properties of the English version of Qualiveen. METHODS: Fifty-five Canadian MS out-patients completed a set of questionnaires, including Qualiveen, MSQOL-54, a MS-specific HRQL questionnaire, urinary function assessments and the Expanded Disability Status Scale (EDSS) twice at an interval of two to four weeks. RESULTS: Qualiveen proved internally consistent (Cronbach's alpha coefficients 0.73 to 0.90 for the four Qualiveen domains) and test-retest reliable (intraclass correlation coefficients 0.88 to 0.94). Consistent with a priori predictions, we found a strong association between overall Qualiveen score and the degree of incontinence (0.63), a moderate correlation with the type of urinary symptoms (0.49), a weak association with manner of voiding (0.28) and weak or absent correlations with MSQOL-54 domains, EDSS bladder/bowel and global EDSS. Predictions proved generally accurate (weighted kappa = 0.65). CONCLUSION: The internal consistency, test-retest reliability and cross-sectional construct validity of the English version of Qualiveen are excellent, and similar to the original French version. Further studies should explore Qualiveen's longitudinal validity and responsiveness. PMID- 15892432 TI - Quality of life and temperament factors in schizophrenia: comparative study of patients, their siblings and controls. AB - This study aimed to determine which temperament factors are associated with quality of life (QOL) in schizophrenia based on a triplet design comparing patients with their non-affected siblings and healthy control subjects. Forty seven DSM-IV clinically stable schizophrenia outpatients, 47 non-affected siblings, and 56 non-patients matched for gender and age were evaluated using the Tridimensional Personality Questionnaire and the Quality of Life Enjoyment and Life Satisfaction Questionaire. As expected, schizophrenia patients reported significantly poorer QOL in most specific domains and in general. They also revealed significantly higher scores on harm avoidance and scored lower on reward dependence, than both their siblings and controls. Tridimensional Personality Questionnaire temperament factors revealed no differences between the controls and the siblings. When differences between patients, their siblings and controls were adjusted for gender, age at examination, and education, ANOVA demonstrated that temperament factors contributed to poor satisfaction with physical health, social relationships and subjective feelings in schizophrenia patients. Harm avoidance was associated with general QOL independent of severity of psychopathology. Thus, this study suggests that temperament factors that are not necessarily part of the deterioration process of the illness are associated with the quality of life of schizophrenia patients. These and other non-illness related factors should be considered when evaluating quality of life outcomes in intervention studies. PMID- 15892433 TI - Changes in quality of life in chronic psychiatric patients: a comparison between EuroQol (EQ-5D) and WHOQoL. AB - It has often been postulated that simple, short questionnaires are unable to reflect complex changes in well-being of individuals with chronic psychiatric disorders. To investigate these assumptions we included two recently developed instruments to measure quality of life (the WHOQoL-Bref and the EuroQoL EQ-5D) in a randomised control trial (RCT) in which two treatment conditions were compared. Aims of the study were to assess the sensitivity and validity of these quality of life (QoL)-instruments, to establish their relationship and to examine the predictors of changes in QoL. Subjective changes in QoL were measured on three assessments waves in a period of 18 months and compared to objective changes in psychopathology and social functioning in a sample of 76 chronic schizophrenic patients who participated in the RCT. Results indicated that both WHOQoL-Bref and EuroQoL EQ-5D are capable of detecting changes in QoL over time in physical and psychological well-being. The instruments partly measure the same aspects of QoL, indicated by 50% common variance on total scores. Reduction of positive psychotic symptoms appeared to be the most important factor in improving QoL. The weighted TTO-score of EuroQoL-5D, which is often used as an index in economic evaluations of health care, did however not correspond with these changes, which indicates that it is less sensitive to changes in social and psychological well-being. It's use as the core measure in (economic) health evaluation in the field of psychiatry therefore seems less appropriate. PMID- 15892434 TI - Quality of life related to type 1 Gaucher disease: Spanish experience. AB - The impact of type 1 Gaucher disease and its therapy on health-related quality of life (QOL) was investigated and the results were compared with a Spanish adult normative group. PATIENTS AND METHODS: Between January 1998 and December 2002, a prospective clinical QOL trial was conducted by application of a Spanish version of the Health Survey SF-36 questionnaire. Patients receiving ERT (69 cases) filled in the questionnaire two times, prior to starting ERT and after two years under ERT. The patients were stratified by gender and age group. Clinical and X ray data to assess bone disease were obtained from the Spanish Gaucher Register. Demographic, clinical, genotype and analytical data and the response to therapy were evaluated. Four grades of severity were established according to bone disease (no symptoms = 0, moderate bone pain = 1, severe bone crisis = 2, fracture/necrosis = 3). Correlation analysis was made between QOL score and grade of bone disease. RESULTS: Mean age+/-SD 33.6+/-11.7 (range 18-66), M/F,ratio 33/36; bone disease: 0 in 27 patients (47.3%), 1 in 11 (19.3%), 2 in 5 (8.8%) and 3 in 14 (24.5%). Physical activity: 11 patients (19.3%) showed severe restriction and 41 patients (71.9%) were only limited for strenuous activities. The mean score for QOL questionnaire was 11.9+/-10.4 (range 2-46). Correlation between score and bone disease was significant only for 1 and 3 grades (p = 0.02). Improvement in self perception of global health was observed ranging from 34.3% before ERT to 91.4% after ERT (p = 0.001). Nevertheless physical activity remained unsatisfactory in 24.5% of patients due mainly to bone sequelae. COMMENTS: Physical activities and bone disease grade 1 and 3 are negatively related to QOL. Nevertheless no correlation was found with bone pain crisis, possibly due to the transitory character of this event. In spite of the improvement induced by ERT, a quarter of patients remained with physical limitations related to bone disease as well as in need of orthopaedic correction of bone sequelae. In order to improve the QOL an accurate evaluation of bone disease to define therapeutic approaches must be considered. PMID- 15892435 TI - Quality of life of caregivers in Parkinson's disease. AB - OBJECTIVE: To assess the impact of PD on informal caregivers of patients and identify the main factors related to caregiver strain. PATIENTS AND METHODS: Pairs of PD patients and their caregivers. Evaluation by neurologists included the Hoehn and Yahr, Schwab and England, UPDRS (parts 1-3), ISAPD, and Pfeiffer's SPMSQ rating scales. Patients completed the Euro-QoL 5D, PDQ-8, and Hospital Anxiety and Depression Scale. The SQLC was used to assess caregivers' quality of life (QoL), with caregivers, in turn, applying the Euro-QoL and PDQ-8 to assess patients' health-related quality of life (HRQoL). Multiple linear regression models were fitted to ascertain factors linked to the SQLC. RESULTS: Significant correlations were in evidence between the following scores: SQLC and clinical rating scales and SQLC and patients' HRQoL. Based on multiple regression analysis, patients' functional state (ADL) proved to be the main predictor of caregivers' QoL. Self- and caregiver-assessed patients' HRQoL also proved to be a relevant factor. CONCLUSIONS: (1) Patients' functional state was significantly related to caregivers' psychosocial burden; (2) patients' HRQoL proved to be an additional factor linked to caregiver QoL; (3) improvement of patient disability and HRQoL might alleviate caregiver strain. PMID- 15892436 TI - Psychometric properties of the World Health Organization quality of life instrument (WHOQoL-BREF) in alcoholic males: a pilot study. AB - OBJECTIVE: To evaluate criterion convergent and discriminant validity of the World Health Organization Instrument (WHOQoL-BREF) in evaluating quality of life and to verify its convergent validity with the Medical Outcomes study Short-forms (SF-36) and the Symptom Check List (SCL-90) instruments in male alcohol dependent patients who seek treatment at two Brazilian treatment centers. METHODS: A cross sectional study was performed, in which cases were divided in to two groups according to the severity of alcohol dependence, ascertained by the Short Form Alcohol Dependence Data scale (SADD). RESULTS: The sample was comprised of 36 males, and 63.9% had severe dependence. Subjects with low/moderate dependence showed higher scores in all domains of the WHOQoL-BREF and in nearly all domains of the SF-36. Criterion validity and internal consistency in the WHOQoL-BREF were satisfactory. With regard to convergent validity between the WHOQoL-BREF and SF 36, most correlation coefficients were significant. CONCLUSIONS: The WHOQoL instrument proved to be satisfactory for evaluating quality of life in this sample. PMID- 15892437 TI - Gender differences in health-related quality of life in patients with HIV/AIDS. AB - BACKGROUND: In studies evaluating the general US population, patients in primary care, and patients with chronic conditions, women consistently report poorer health-related quality of life (HRQoL) than men; however, studies evaluating HRQoL in patients with HIV/AIDS have not completely corroborated those findings. The objective of this study was to evaluate gender differences in HRQoL for participants in a large randomized trial comparing antiretroviral regimens. METHODS: AIDS Clinical Trials Group (ACTG) 320 was a randomized, blinded, placebo controlled trial comparing the 3-drug regimen of indinavir + zidovudine (or stavudine) + lamivudine with the 2-drug combination of zidovudine (or stavudine) + lamivudine in subjects with CD4 cell counts less than 200 cells/microl and no prior treatment with protease inhibitors. Nine quality of life domains scored on 0-100 scales were assessed using the ACTG QOL 601-602 Health Survey at 3 points in the trial: baseline, 24 weeks and 40 weeks. Differences between men and women in HRQoL scores were assessed using the Wilcoxon rank-sum test and generalized estimating equation (GEE) models. RESULTS: Overall, 202 females and 976 males were randomized to one of two treatment arms. Female participants were more likely to be black or Hispanic and tended to be younger. At baseline, females reported lower HRQoL scores than males in all of the domains except social functioning, and at week 40, women scored lower in all of the domains except overall health. In repeated measures models, women were found to score lower in all HRQoL domains except overall health, with significant differences of 3.5-6.7 points in 3 of the 9 quality of life domains: physical functioning, pain, and energy/fatigue. HRQoL scores improved for participants in the study over time and in response to potent treatment, and the improvements were similar for men and women. CONCLUSIONS: Women with HIV/AIDS report substantially poorer HRQoL than men with HIV/AIDS in several HRQoL domains. However, changes in domain scores over time and in response to treatment do not differ significantly by gender, implying that changes in domain scores may be better HRQoL outcomes to compare between HIV-infected men and women in clinical trials than mean domain scores. PMID- 15892438 TI - Effect of order of administration of health-related quality of life interview instruments on responses. AB - PURPOSE: To determine the effect on patient responses from the order in which the generic health-related and vision-targeted instruments are administered in a set of randomized clinical trials of intraocular surgery. PATIENTS AND METHODS: Patients who agreed to enroll in the Submacular Surgery Trials (SST) completed baseline quality of life interviews prior to random assignment to surgery or observation. Interviews were conducted by trained interviewers located at the SST Coordinating Center, via a computer-assisted telephone interview system that randomly assigned the order of instrument administration. Either the generic health-related instruments were administered first, the SF-36 Health Survey (SF 36) followed by the Hospital Anxiety and Depression Scale (HADS), followed by the vision-targeted instruments, National Eye Institute-Vision Function Questionnaire (NEI-VFQ) followed by the SST-Vision Preference Value Scale, or the vision targeted questions were asked first, followed by the generic health instruments. The four instruments have 25 subscales total. RESULTS: Of the 1015 patients enrolled in the SST, 992 patients had all four instruments administered in random order: 483 (49%) patients responded to the generic instruments first and 509 (51%) patients responded to the vision-targeted instruments first. Order of administration produced significantly different scores for three health status subscales: SF-36 mental health, HADS depression and HADS anxiety (p < or = 0.05, Wilcoxon rank sum test). CONCLUSIONS: Overall, the order of administration did not have a large effect on responses to the baseline interviews in this study. However, three mental health subscales were affected by order, though the order effect was small in magnitude. When the generic health instruments followed the vision-targeted instrument the HADS depression and anxiety scores were higher and the SF-36 mental health scores were lower, both suggesting poorer mental health status. Thus, the order of administration in other settings in which both a generic health-related instrument and a condition-targeted instrument are used may be decided based on individual study goals and priorities but order of administration should be consistent throughout the study. PMID- 15892439 TI - Health-related quality of life during the bereavement period of caregivers of a deceased elderly person. AB - Few studies have addressed the health-related quality of life (HRQL) impact of bereavement. The objective of this study was to evaluate the HRQL of the main caregiver after the death of a close elderly relative. 142 caregivers were invited to participate in a phone interview with the SF-36. General population SF 36 norms, were used to calculate the gender and age standardised physical component summary (PCS) and mental component summary (MCS) scores. A MCS below 43 was used as an indicator of possible depression. Databases of several patient studies were additionally used to facilitate interpretation of results. 130 caregivers (91.5%) participated, the majority being women (80%), middle aged (56 years), and the spouse (22%) or son/daughter (56%) of the deceased person. Caregivers' SF-36 scores were lower than expected for mental health, role emotional and social functioning. Their MCS was below that expected by 0.6 SD units, whereas their PCS was above by 0.2 SD. The proportion of caregivers with possible depression was 33.8%. First-degree family relationship with the deceased (OR = 4.5), and lower age of the deceased (OR = 3.8) were significantly associated (p < 0.02) to a possible depression. The recent death of an elderly person is associated with significant emotional health impact on the caregiver. PMID- 15892440 TI - Social network and health-related quality of life in older adults: a population based study in Spain. AB - This study examined the association between social network and health-related quality of life (HRQL) in older adults and compared this against the association between HRQL and a disabling disease such as osteoarthritis. A cross-sectional survey was done on 3600 subjects representative of the Spanish non institutionalised population aged 60 years and over. Data were collected through home-based personal interview and physical examination. HRQL was measured with the SF-36 health questionnaire. Data analysis was performed with multiple linear regression models with adjustment for the main confounders. Of the total sample, 38.6% of subjects were unmarried, 17.6% were living alone, 4.7% saw their family seldom or never, and 2.9% saw their friends seldom or never. Unmarried status and living alone were associated with lower scores in the social and mental quality of-life components, though statistical significance was not in general attained (p > 0.05). Seeing family members seldom or never was associated (p < 0.05) with worse scores in the following scales of SF-36 questionnaire: role-physical, body pain, general health and mental health. HRQL was lower among those who saw friends seldom or never, and the reduction in HRQL proved similar to that associated with osteoarthritis, on the physical functioning (coefficients -8.4 vs. -8.1) and general health scales (-7.8 vs. -6.6); the reduction in HRQL was even greater than that associated with osteoarthritis for other scales, such as vitality (-9.6 vs. -6.7; p > 0.05) and social functioning (-14.5 vs. -3.7; p < 0.05). We conclude that only a small proportion of Spain's elderly population lack frequent social relationships, yet low frequency of relationships with friends is associated with a decline in quality of life similar to or greater than that associated with osteoarthritis. PMID- 15892441 TI - Quality of life of participants in a mind-body-based self-development course: a descriptive study. AB - OBJECTIVE: A cross-sectional survey of quality of life of people attending a self development course involving psychophysiological mind-body medicine (MBM) activities. DESIGN: A questionnaire study using a health-related quality of life (HRQoL) instrument, the SWED-QUAL, with 13 subscales scored 0-100, and questions about utilisation of alternative and standard health care, medication and sick leave. SETTING: A training centre for MBM, established 15 years ago. STUDY GROUP: One hundred and seven eligible course attendants (response rate 88%, age 20-70 years) during the year 2000 assessed their HRQoL just before entering the course. Attendance was self-initiated, without referral. The results on HRQoL were compared with those of control subjects from the general Swedish population. RESULTS: Six of the thirteen HRQoL subscales were strongly and significantly reduced (p < 0.0001) in the study group: Negative affectivity, Role limitation due to emotional health, Positive affectivity, Cognitive functioning, Family functioning and Marital functioning. Long-term sick leave (>6 months) was three times as frequent in the study group as in the general population. Use of psychotropic medication was slightly increased compared to the general population, at least among the younger male participants. The education level was high, health care utilisation was average and body functioning was good. CONCLUSIONS: This group of well-educated men and women gave their emotional health an unexpectedly low rating, on a par with that given by people with chronic diseases. PMID- 15892442 TI - Does being bilingual in English and Chinese influence changes in quality of life scale scores? Evidence from a prospective, population based study. AB - BACKGROUND: Bilinguals differ from monolinguals in language use, but the influence of bilingualism on changes in Health-Related Quality of Life (HRQoL) scores is not known. OBJECTIVE: To determine the influence of bilingualism on changes in HRQoL scores. RESEARCH DESIGN: A prospective cohort study of a population-based, disproportionately stratified random sample of monolingual or bilingual ethnic Chinese who completed the Short-Form 36 Health Survey (SF-36) in English or Chinese twice in 2 years. Least squares regression models were used to assess the influence of bilingualism on SF-36 scores, while adjusting for the influence of questionnaire language and determinants of HRQoL. RESULTS: Usable English and Chinese questionnaires were returned by 1013 and 910 subjects respectively (aged 21-65 years, 48.5% female, 52.8% bilingual). Bilinguals differed from monolinguals in known determinants of HRQoL (being younger and better educated), changes in determinants of HRQoL over 2 years (more bilinguals had changes in work or marital status) and had mean SF-36 scores that were up to 10 points higher than monolinguals. After adjusting for these differences, bilingualism did not influence 2 year change scores for any of 8 SF-36 scales. CONCLUSION: Bilingualism did not influence changes in HRQoL scores over 2 years in this large, population-based study of subjects fluent in English and/or Chinese (representing an alphabet and/or pictogram based language respectively). PMID- 15892443 TI - Is the standard SF-12 health survey valid and equivalent for a Chinese population? AB - INTRODUCTION: Chinese is the world's largest ethnic group but few health-related quality of life (HRQoL) measures have been tested on them. The aim of this study was to determine if the standard SF-12 was valid and equivalent for a Chinese population. METHODS: The SF-36 data of 2410 Chinese adults randomly selected from the general population of Hong Kong (HK) were analysed. The Chinese (HK) specific SF-12 items and scoring algorithm were derived from the HK Chinese population data by multiple regressions. The SF-36 PCS and MCS scores were used as criteria to assess the content and criterion validity of the SF-12. The standard and Chinese (HK) specific SF-12 PCS and MCS scores were compared for equivalence. RESULTS: The standard SF-12 explained 82% and 89% of the variance of the SF-36 PCS and MCS scores, respectively, and the effect size differences between the standard SF-36 and SF-12 scores were less than 0.3. Six of the Chinese (HK) specific SF-12 items were different from those of the standard SF-12, but the effect size differences between the Chinese (HK) specific and standard SF-12 scores were mostly less than 0.3. CONCLUSIONS: The standard SF-12 was valid and equivalent for the Chinese, which would enable more Chinese to be included in clinical trials that measure HRQoL. PMID- 15892444 TI - Translation and psychometric testing of the Basque version of the SF-36 health survey. AB - INTRODUCTION: The primary goals of this study were to adapt the Short Form-36 (SF 36) questionnaire to the Basque language and to assess the translated questionnaire's psychometric properties. METHODS: Two pilot studies and a field study were conducted in 1999. The pathologies selected were hypertension, eating disorders, back pain, and HIV infection; blood donors were also included. The analyses conducted at item-level were: item internal consistency, item discriminant validity, equal item variance, and equal item-scale correlation. Internal consistency was measured at the scale level. Reproducibility, convergent validity and discriminant validity were also examined. RESULTS: 285 individuals took part in the study; sufficient data were obtained for 265 (93%). All items correlated higher than 0.4 with their hypothesized scales. Within each scale, item standard deviations were similar. Item-scale correlations were also similar. Cronbach's alpha coefficients varied from 0.76 to 0.92. In the test-retest reliability study, which included 54 additional individuals, all scales were above 0.50. Regarding the inter-scale correlation, the majority exceeded the 0.40 coefficient. In the correlation of similar scales of the SF-36 and the Nottingham Health Profile, intraclass correlation coefficient results ranged from 0.29 to 0.62. CONCLUSIONS: These results confirm that this new version of the SF-36 has been translated and adapted correctly for the Basque language and that it fulfils, at least partially, the psychometric properties required for this instrument. Further studies, however, are needed to completely validate the Basque version of the SF-36. PMID- 15892445 TI - Discriminative ability of the short-form 36 health survey: a tale of two versions. AB - BACKGROUND AND OBJECTIVES: Previous studies suggested that the Chinese version of the Short-Form 36 Health Survey (SF-36) had smaller variability in the physical functioning (PF) and physical component summary (PCS) scores than the English SF 36. This may translate into differences in discriminative ability and sample size requirement. MATERIALS AND METHODS: Data were drawn from a community-based survey and a randomised crossover study of Singaporeans bilingual in the Chinese and English languages. The abilities of the two PF and PCS versions in discriminating subjects who reported chronic illness and acute disease symptoms versus those who did not were compared. RESULTS: In all four comparisons (i.e. two health criteria in two studies) the Chinese version of PF showed a larger effect size than the English version. In three out of four comparisons the Chinese version of PCS showed a larger effect size than the English version. CONCLUSIONS: The Chinese version appeared more efficient in detecting a statistically significant difference between groups. Other factors being the same, the Chinese SF-36 may require a smaller sample size than the English SF-36 for the studies of physical aspects of health-related quality of life. PMID- 15892446 TI - Reliability and validity of the WHOQOL BREF in a sample of Brazilian outpatients with major depression. AB - The present study assessed the psychometric properties of the Brazilian version of the World Health Organization's Quality of Life Instrument--Short Version (WHOQOL BREF) in a sample of 89 adult outpatients with major depression. After analyses, the WHOQOL BREF showed good internal consistency, and was sensitive to improvement after treatment with antidepressants. Convergent validity between the WHOQOL BREF and the Beck Depression Inventory was statistically significant, as well as WHOQOL BREF's ability to discriminate between outpatients on the basis of their level of depression. In conclusion, the WHOQOL BREF seems to be a psychometrically valid and reliable instrument that it is suitable for evaluating the quality of life of Brazilian-speaking depressed outpatients. PMID- 15892447 TI - Reliability and validity testing of a Chinese-translated version of the 39-item Parkinson's Disease Questionnaire (PDQ-39). AB - The trend toward international cooperation in research projects emphasizes the need to translate existing validated tools into local languages. The purpose of this study was to test the reliability and validity of a Chinese-translated version of the 39-item Parkinson's Disease Questionnaire (PDQ-39). Seventy-three Taiwanese individuals with Parkinson's disease were consecutively recruited. The internal consistency reliability was satisfactory for all domains (Cronbach's alpha = 0.80-0.96), except for the social support, cognition, and bodily discomfort domains (alpha = 0.58-0.63). The convergent validity was also supported by strong correlations between domains measuring related constructs of the PDQ-39 and Unified Parkinson's Disease Rating Scale (r = 0.81-0.86), and between those of the PDQ-39 and SF-36 (r = -0.70--0.93). Except for the bodily discomfort domain, all domains of the PDQ-39 significantly discriminated patients at different stages, as indicated by the Hoehn and Yahr scale. Overall, the results of this study are consistent with the reports of the PDQ-39 in other countries (e.g., UK, Spain, US, and Greece), which suggests that the PDQ-39 is appropriate for use among Taiwanese individuals with PD. This study lays the foundation for future combinations and comparisons of data cross-nationally. PMID- 15892448 TI - The transcriptional response to hypoxic insult controlled by FRA-2. AB - FRA-2 is involved in cellular differentiation and is also upregulated in response to ischemic injury to the brain. To shed light on the function of this transcription factor, a novel microarray analysis was utilized to identify FRA-2 dependent gene expression increased in the hypoxic response. Genes were identified that were upregulated by exposure of neuronally differentiated PC12 cells to hypoxia. Using a dominant negative construct to block FRA-2, a second subset of genes that were FRA-2 dependent was found. Cross comparison then allowed isolation of a list of genes that were induced in response to hypoxia in a FRA-2-dependent manner. These data suggest that FRA-2 is involved in the transcriptional control of neuroprotective genes and in the switch from aerobic to anaerobic metabolism. PMID- 15892449 TI - Quantification of G protein Gaalphas subunit splice variants in different human tissues and cells using pyrosequencing. AB - The G protein Galphas is derived from four alternatively spliced transcripts, two long variants (Galphas(L)+CAG and Galphas(L)-CAG), which include an extra 45-bp segment, and two short variants (Galphas(S)+CAG and Galphas(S)-CAG). The long and short forms differ in each case by splicing in or out of a serine residue encoded at the 3' end of the variable exon 3. The relative expression of all four variants in human tissues is poorly investigated due to experimental limitations. We therefore established a method for reliable relative mRNA quantification of these splice variants based on the Pyrosequencing technology, and determined Galphas transcript ratios in various human tissues and cells. Galphas(S)/Galphas ratio was highest in blood mononuclear cells (0.84 +/- 0.02, n = 16) and lowest in the brain (0.51 +/- 0.14, n = 3). The different ranges resulted from differences in Galphas(S)+CAG ratios, which ranged from a total Galphas ratio of 0.32 +/- 0.07 (n = 12) in heart tissue to 0.57 +/- 0.03 (n = 16) in blood mononuclear cells (p < 0.0001), whereas the Galphas(S)-CAG ratio was rather constant and ranged from 0.22 +/- 0.04 (n = 7) in retinoblastoma cells to 0.27 +/ 0.04 in lymphocytes (p = 0.19). The Galphas(L)+CAG ratio ranged from 0.02 +/- 0.02 in heart tissue to 0.05 +/- 0.01 in retinoblastoma cells, with a varying proportion of Galphas(L)-CAG, which ranged from 0.14 +/- 0.02 in blood mononuclear cells to 0.41 +/- 0.08 in heart tissue. Stimulation of immortalized B lymphoblasts with isoproterenol resulted in significant changes of splice variant ratios. Our data indicate that changes of long and short ratios of Galphas in different tissues affected Galphas(L)-CAG and Gas(S)+CAG rather than Galphas(L)+CAG and Galphas(S-)CAG. Furthermore, stimulation of cells seemed to affect splice variant ratios. These results are, therefore, suggestive of different biological functions of these variants. PMID- 15892450 TI - Expression of progenitor cell markers during expansion of sorted human pancreatic beta cells. AB - Functional pancreatic beta cell mass is dynamic and although fully differentiated, beta cells are capable of reentering the cell cycle upon appropriate stimuli. Stimulating regeneration-competent cells in situ is clearly the most desirable way to restore damaged tissue. Regeneration by dedifferentiation and transdifferentiation is a potential source of cells exhibiting a more developmentally immature phenotype and a wide differentiation potential. In this context and to gain a better understanding of the transformation induced in human beta cells during forced in vitro expansion, we focused on identifying differences in gene expression along with phenotypical transformation between proliferating and quiescent human beta cells. FACS purified beta cells from three different human pancreata were cultured during 3-4 months (8-10 subcultures) on HTB-9 cell matrix with hepatocyte growth factor. Gene expression profiling was performed on cells from each subculture on "in house" pancreas-specific microarrays consisting of 218 genes and concomitant morphological transformations were studied by immunocytochemistry. Immunocytochemical studies indicated a shift from epithelial to neuroepithelial cell phenotype, including progenitor cell features such as protein gene product 9.5 (PGP 9.5), Reg, vimentin, and neurogenin 3 protein expression. The expression of 49 genes was downregulated, including several markers of endocrine differentiation while 76 were induced by cell expansion including several markers of progenitor cells. Their pattern also argues for the transdifferentiation of beta cells into progenitor cells, demonstrating neuroepithelial features and overexpressing both PBX1, a homeodomain protein that can bind as a heterodimer with PDX1 and could switch the nature of its transcriptional activity, and neurogenin 3, a key factor for the generation of endocrine islet cells. Our study of the machinery that regulates human beta cell expansion and dedifferentiation may help elucidate some of the critical genes that control the formation of adult pancreatic progenitor cells and hence design targets to modify their expression in view of the production of insulin-secreting cells. PMID- 15892451 TI - Translational downregulation of the noncatalytic growth factor receptor TrkB.T1 by ischemic preconditioning of primary neurons. AB - Short episodes of ischemia can protect neuronal cells and tissue against a subsequent lethal ischemia by a phenomenon called ischemic preconditioning. The development of this tolerance depends on protein synthesis and takes at least 1 day. It therefore seems reasonable that preconditioning leads to upregulation and translation of protective genes or posttranslational modification of pro- or antiapoptotic proteins. We recently used suppression subtractive hybridization to identify transcripts upregulated in rat primary neuronal cultures preconditioned by oxygen glucose deprivation. In this contribution, we describe the previously unknown 7-kb full-length sequence of an upregulated expressed sequence tag and show that it constitutes the 3' end of the large untranslated region of the noncatalytic "truncated" growth factor receptor TrkB.T1. TrkB.T1 is expressed most prominently in the adult brain and its mRNA was found to be 2.1-fold upregulated by ischemic preconditioning. At the protein level, however, TrkB.T1 was clearly downregulated, possibly by increased degradation in preconditioned cultures. TrKB.T1 can act as a dominant-negative inhibitor of its catalytic counterpart TrkB, which is the receptor for brain-derived neurotrophic factor (BDNF), a factor induced by ischemia that can protect from ischemia-induced neuron loss. We hypothesize that the downregulation of TrkB.T1 at the protein level can prolong BDNF-mediated protective signaling via the catalytic receptor and thus participates in the development of ischemic preconditioning. PMID- 15892452 TI - Exercise-induced gene expression changes in the rat spinal cord. AB - There is growing evidence that exercise benefits recovery of neuromuscular function from spinal cord injury (SCI). However, the effect of exercise on gene expression in the spinal cord is poorly understood. We used oligonucleotide microarrays to compare thoracic and lumbar regions of spinal cord of either exercising (voluntary wheel running for 21 days) or sedentary rats. The expression data were filtered using statistical tests for significance, and K means clustering was then used to segregate lists of significantly changed genes into sets based upon expression patterns across all experimental groups. Levels of brain-derived neurotrophic factor (BDNF) protein were also measured after voluntary exercise, across different regions of the spinal cord. BDNF mRNA increased with voluntary exercise, as has been previously shown for other forms of exercise, contributed to by increases in both exon I and exon III. The exercise-induced gene expression changes identified by microarray analysis are consistent with increases in pathways promoting neuronal health, signaling, remodeling, cellular transport, and development of oligodendrocytes. Taken together these data suggest cellular pathways through which exercise may promote recovery in the SCI population. PMID- 15892453 TI - Transcriptional profile of NeuroD expression in a human fetal astroglial cell line. AB - NeuroD1, a member of the basic helix-loop-helix (bHLH) protein family, is a transcription factor that plays a pivotal role in terminal differentiation of neural progenitors. The primary objective was to generate an early transcriptional profile triggered by NeuroD1 to guide future studies on mechanisms of neuronal differentiation. The human NeuroD1 coding region was amplified from human fetal brain RNA using specific primers and cloned into a CMV expression vector (CT-GFP-TOPO/pcDNA3.1). Transfection of a fetal glial cell line with this construct resulted in expression of NeuroD1 in 13-15% of the cells. Markers typical of early neuronal development were observed by immunocytochemical staining in a small proportion of transfected cells. To enrich the population of NeuroD1-expressing cells, fluorescence-activated cell sorting (FACS) was used to purify and collect the NeuroD1/GFP+ cells. Total RNA was extracted from the pair of cultures (NeuroD1/GFP vs. control plasmid/GFP) and processed for gene expression studies. A final gene list was composed from those probe sets that were either increased or decreased in the NeuroD1-expressing cells in three independent experiments (p < 0.001). Each gene was investigated further for possible roles in neurogenesis and a subset of 177 genes was chosen based on the following characteristics: a) genes that are potential NeuroD1 dimerization partners, b) genes that modulate other bHLH transcription factors, c) genes related to development, and d) genes associated with neural induction, outgrowth, and terminal differentiation. DNA microarray analysis of NeuroD1 expression in an astroglial cell line produced a "snapshot" transcriptional profile that will be useful in deciphering the complex molecular code that specifies a neuronal fate. PMID- 15892454 TI - [The meaning of teaching history of medicine in surgery residency programmes]. PMID- 15892455 TI - [Current concepts on sudden death]. AB - Sudden death is defined as the death occurring less than one hour before the onset of the patient's symptoms. It is a severe condition considered a public health issue in several countries and in ours, it accounts for 33 000 to 53 000 annual deaths mainly related to ischemic heart disease. The main cause of sudden death are severe ventricular arrhythmias, but determining what patients are at risk for such an episode is complex, that is why risk stratification is usually a low cost-effective intervention. In the present study, we describe different sudden death risk-stratification strategies. Different sudden death treatment strategies regarding general population have different success rates in different countries, nevertheless, among select high risk populations; the best therapy currently available is the automatic implantable cardioverter-defibrillator. We also discuss other treatment options. In Mexico it is deemed necessary to do an important effort for the early detection, prevention and treatment of sudden death in order to limit the consequences of this problem. PMID- 15892456 TI - [Breastfeeding counseling and early mother-child contact are associated with exclusive maternal breastfeeding. A hospital-based-case-control study]. AB - BACKGROUND: The aim of this study was to assess the association between exclusive breastfeeding (EBF) and some factors that may influence breastfeeding in a closely related population attending a private hospital sponsored by a major Mexican brewing company. MATERIALS AND METHODS: We carried out a retrospective hospital-based unmatched case-control study. A sample of 124 mother-newborn couples was interviewed in a private medical unit in Monterrey, Mexico, from January 2001 to January 2002. The association between EBF and 11 explanatory factors was analyzed by logistic regression analysis. RESULTS: Counseling and early contact between mother-newborn couples were positively associated with EBF. CONCLUSION: Counseling and early contact improve EBF practice. We believe these two practices would favor better adherence to EBF in other populations. PMID- 15892457 TI - [Disability due to auditory and vestibular dysfunction in a specialized care center]. AB - OBJECTIVE: To assess the frequency and characteristics of the disability associated with hearing loss and vestibular disease in a Specialized Medical Center of the Mexican Social Security Institute. METHODS: 530 patients agreed to participate. They were assessed due to hearing loss (n = 252) and vestibular disease (n = 278), 54% and 50% of them worked. After a clinical evaluation and administration of a symptom questionnaire, they were asked about the frequency and days ofdisability to perform daily life activities related to their audiological or vestibular disease and how often they visited the physician during the last year because of their symptoms. RESULTS: Hearing loss patients visited the physician 1-6 times/ year and 15.8% reported disability. Vestibular patients visited the physician 1-8 times/ year and reported disability more frequently (60.8%) (p < 0.01). Among those who worked, 5% of patients with hearing loss and 51% of patients with vestibular disease stopped working during 1 15 cumulative days for the first group and 1-365 cumulative days for the second group. Spearmnan's correlation coefficient between the evolution of the disease and disability days was -0.14 (p < 0.01) for hearing loss patients and -0.27 (p < 0.01 ) for vestibular disease. CONCLUSION: Vestibular disease is a cause of disability. which can have financial impact on both the patient and the health care system. PMID- 15892458 TI - [Inconsistency in sun protection factor (SPF) index in Mexico. The case of sunscreens for oily skin]. AB - BACKGROUND: In Mexico, information regardiing sunscreen protection is not widely accessible from sources other than manufacturers. OBJECTIVE: To assess the Sun Protection Factor (SPF) of 12, over the counter sunscreens for oily or acne-prone skin available in the Mexican market. SETTING: Dermatology Department. Autonomous University of San Luis Potosi, Mexico. SUBJECTS AND METHODS: Twenty healthy volunteers ofskin type III and IV were tested. Sunscreen SPFs were measured using solar simulated radiation according to FDA final monograph. RESULTS: SPFs ranged between 7.8 and 26.9. Overall SPFs determined by solar simulator showed values between 22% to 74% less than the one advertised in their labels. CONCLUSIONS: We warn about the potenital risk of SPF overestimation as many of'the sunscreens tested did not comply with their offered protection. A proper regulation is needed because sunscreens are considered cosmetic products and do not require clinical tests to verify their efficacy before marketing. PMID- 15892459 TI - [Identifying different susceptibility loci associated with early onset diabetes and cardiovascular disease in Mexican families]. AB - Coronary artery disease and diabetes mellitus are among the primary mortality and morbidity causes in Mexico. Genetic factors play a fundamental role in the development of these entities. In the past few years due to the recognition and study of families with monogenic forms of diabetes and dislipidemias associated with development of atherosclerosis, several genes and loci have been associated with these conditions through genetic linkage studies. These studies have provided evidence of the genetic heterogeneity that exists and the type of genes involved in different ethnic groups. The study of Mexican families with early onset diabetes and combined familial hyperlipidemia showed the participation of different genetic loci associated with these conditions in the Mexican population. These findings show the value of gene mapping strategies in the identification of the genetic component in these entities in our population. PMID- 15892460 TI - [Acute pancreatitis: five year experience at the Hospital General de Mexico]. AB - INTRODUCTION: In Mexico acute pancreatitis is an important cause of morbility and mortality. We here describe the characteristics of patients managed with that diagnosis at the Hospital General de Mexico. MATERIAL AND METHODS: Retrospective, descriptive and observational study. Patients with acute panrcreatitis were seen between January 1996 to December 2000. RESULTS: The acute pancreatitis prevalence found in the hospital setting was 5%. We identified 104 patients, 62% men, with mean age 37 yrs. In 74% ofcases seen, we made a correct early diagnosis upon admission. The biliary (49%) and the alcoholic (37%) were the most frequent causes. Abdominal pain was the most common symptom at onset (94%). High leukocyte count (78%), followed by serum lipase (74%) was the most sensitive laboratory test. Only 34% of patients studied had an adequate nutritional status. 54% of' patients presented severe pancreatitis. The mean number of hospital days was 17 and 44% required ICU management. Thirty five percent required surgery where necrosectomy was performed and a mortality of 67% was found. The total mortality rate was 21%. CONCLUSIONS: The correct clinical diagnosis of acute pancreatitis is frequent. It is a more common finding than severe pancreatitis, but it does not increase the morbility and mortality in our patient population. PMID- 15892461 TI - [Reflections on the quality of medical school curricula in Mexico]. AB - INTRODUCTION: At the end of the XXth century, collegiate efforts (evaluation and accreditation) were carried out to identify the quality of undergraduate medical education in Mexico. The growth of medical schools during the 1970's had stabilized and the combination of other factors had contributed to increase its quality. MATERIAL AND METHODS: Analytic-descriptive research of selected indicators of the medical curricula from the sixties to the year 2000: entry, enrollment, and drop out rates, graduates, title, selection by the national postgraduate exam of medical residences (ENARM), faculty members and creation of medical schools. Statistical analysis included chi2 and T tests. RESULTS: In the seventies the number of medical schools doubled and the country increased its entry by 400%, enrollment by 700% and the number of graduates by 600%. The drop out rates and title efficiencies remained stable with averages of 76.2 and 76.4%, respectively: the first one that reflects the efficiency of the educational system, was not different in the period studied, when student's t- test (p < 0.01) was used. In the nineties, entry, enrollment and graduates increased again by 300, 400 and 700%, compared with the seventies. One out of every two graduates was selected by the ENARM in 1991 and in 2001, one out of six. CONCLUSIONS: The growth in enrollment and graduates once again, stands in contrast with the drop out rate that has remained stable. This suggests that the country has not increased significantly the quality of its medical education. The latter could be verified for each institution with the indicators used in the present study and by the assessment and accreditation of the programs. PMID- 15892462 TI - [Autism]. AB - Autism is a static, neuro-developmental disorder, that is life-long and comprises a wide range of behavioral disorders. It has a prevalence of 1-2 per 1,000 with a broad range of severity. Multiple causes have been put forward and genetics plays the major role. According to DSM-IV the main clinical manifestations include impaired social skills, language and non-verbal communication and changes in range of interests and activities. Cognitive deficits are frequent but not universal. Perseveration, affective flatness and lack of insight into other people's thinking and feelings is conspicuous. The neurobiological basis of autism is unknown, but histopathological and biochemical findings suggest a diffuse disorder of neuronal development, most severe in the fronto-temporal cortex, some limbic structures and in the cerebellum. At a biochemical level, abnormalities in serotonin and other neurotransmitters have been reported. PMID- 15892463 TI - [A forty-three year old female with functional deficit, resting dyspnea and right hemitorax pain]. PMID- 15892464 TI - [Pneumoscrotum after colon surgery]. PMID- 15892465 TI - [Proximal pancreatectomy in a case of nesidioblastosis]. AB - A 19 moth-old child who presented seizures secondary to intractable hypoglycemia, fulfilling the clinical and biochemical criteria for hyperinsulinism was studied. Histopathological findings of the pancreas showed the presence of small clusters of b cell islets throughout acinar tissue near ducts, in both the head and the proximal third of the body. Proximal pancreatectomy (60%) and distal pancreatic jejunostomy (Roux-in-Y) were performed. This procedure was effective in reverting hypoglycemia and constitutes the first successful alternative treatment. PMID- 15892466 TI - [The beginnings of trauma management in Mexico City. 1847-1914]. AB - We describe how surgical trauma was managed in Mexico City from 1847 to 1912. Management included minimal interventions, generally ending with the patient's death. Other more radical procedures such as abdomen trauma proposed in 1874 anld performed in 1898 are also described. A variety of techniques employed and type of trauma cases registered that yielded more radical attitudes that brought about the creation of public emergency rooms (the Cruz Verde) and military surgical rooms in railroad cars that oversaw the care of soldiers during the revalutionary period are discussed. PMID- 15892467 TI - [Observation of the ventricular system and subarachnoid space in the skull base by flexible neuroendoscopy: normal structures]. AB - Worldwide in the last two decades, in Latin America in the last decade, and Mexico in particular cerebral endoscopy as part of the minimally invasive neurological surgery, has constituted a significant advance for the treatment of neurological diseases such as congenital or acquired hydrocephalus, Chiari malformation, neurocysticercosis, stroke, ventricular and paraventricular tumors and cysts, arachnoidal cyst and hydrocephalus secondary to shunt malfunction. The lateral ventricles, the third ventricle, the cerebral aqueduct, the fourth ventricle, and the subarachnoid basal cisterns are among the most non-accessible regions of the central nervous system. Due to light-rand fiber optic-systems in combination with the experiences of nine consecutive year's and more than 600 endoscopies, most of them flexible neuroendoscopies, we present the neuroendoscopic approach with flexible cerebral endoscope to the anatomy and its normal variants of the whole ventricle system and the subarachnoid basal cisterns including the subarachnoid space of the cervical spine. We also describe their site related structures including veins, arteries, cranial nerves, pituitary gland, choroidal plexus and their normal anatomic variants. PMID- 15892468 TI - [Interfering RNA: A new therapeutic strategy]. PMID- 15892469 TI - [Bush does not promote world health]. PMID- 15892470 TI - [Social reconstruction can be the best healing after the tsunami. Residents of disaster areas must receive help based on local conditions]. PMID- 15892471 TI - [Towards individual-based follow up of health care results]. PMID- 15892472 TI - [Chronic granulomatous disease--not only the lack of oxygen radicals]. AB - Chronic granulomatous disease (CGD) is caused by mutations in the phox-family of superoxide ion generating enzymes. The clinical manifestations of CGD include increased infection susceptibility, a lupus like dermatitis and inflammatory bowel disease. The severe consequences of this rare disorder need to be handled by specialists, experienced in the care of CGD patients. Many CGD manifestations are due to the lack of superoxide ions, but some also to deficient pumping of protons to the extracellular space. An excessive inflammatory component, e.g. the granuloma formation in various organs, might be secondary to impaired superoxide ion dependent inactivation of inflammatory mediators. CGD can be successfully treated by hematopoietic stem cell transplantation, and gene therapy might become an alternative in the future. PMID- 15892473 TI - [Choose midline incision for the best access to the abdominal cavity]. AB - A midline incision or a wide transverse incision offers good access to the abdominal cavity. A midline incision should probably be preferred. Then no major nerve, muscle or vessel is severed and thereby subsequent neural or abdominal wall dysfunction may be avoided. Both incisions seem to be associated with similar pain, although a subcostal incision may be advantageous for gallbladder surgery. Respiratory function may be better with transverse incisions but there is no randomised study comparing incisions when a postoperative epidural is used. Wound dehiscence rates have not been proven to differ and the rate of incisional hernia is similar with both types of incision. PMID- 15892474 TI - [Nipah virus--another product from the Asian "virus factory"]. PMID- 15892475 TI - [Computer-assisted blood glucose control in intensive care units patients]. PMID- 15892476 TI - [Is accepting an invitation to a conference on Hawaii unethical?]. PMID- 15892477 TI - [The producer-gas epoch in Sweden 1939-1945. A controversial poisoning epidemic laid basis for occupational medicine]. PMID- 15892478 TI - [Comrades! There must be some blood order in the field of hypertension!]. PMID- 15892479 TI - [Drug committees are necessary, according to the SNS reference group]. PMID- 15892480 TI - [MSF prior to the Medical Union's delegates meeting: The Medical Union should extend its work against discrimination]. PMID- 15892481 TI - [Antidepressive treatment is the best prevention against suicide among young people]. PMID- 15892482 TI - [Late skin infections following tsunami injuries]. PMID- 15892483 TI - [The first successful heart transplantation in Sweden]. PMID- 15892484 TI - [Divergent studies results and a "magical" limit]. PMID- 15892485 TI - Research development in homeopathy: a question of methods and priorities. PMID- 15892486 TI - Adjunctive homeopathic treatment in patients with severe sepsis: a randomized, double-blind, placebo-controlled trial in an intensive care unit. AB - BACKGROUND: Mortality in patients with severe sepsis remains high despite the development of several therapeutic strategies. The aim of this randomized, double blind, placebo-controlled trial was to evaluate whether homeopathy is able to influence long-term outcome in critically ill patients suffering from severe sepsis. METHODS: Seventy patients with severe sepsis received homeopathic treatment (n = 35) or placebo (n = 35). Five globules in a potency of 200c were given at 12h interval during the stay at the intensive care unit. Survival after a 30 and 180 days was recorded. RESULTS: Three patients (2 homeopathy, 1 placebo) were excluded from the analyses because of incomplete data. All these patients survived. Baseline characteristics including age, sex, BMI, prior conditions, APACHE II score, signs of sepsis, number of organ failures, need for mechanical ventilation, need for vasopressors or veno-venous hemofiltration, and laboratory parameters were not significantly different between groups. On day 30, there was non-statistically significantly trend of survival in favour of homeopathy (verum 81.8%, placebo 67.7%, P= 0.19). On day 180, survival was statistically significantly higher with verum homeopathy (75.8% vs 50.0%, P = 0.043). No adverse effects were observed. CONCLUSIONS: Our data suggest that homeopathic treatment may be a useful additional therapeutic measure with a long-term benefit for severely septic patients admitted to the intensive care unit. A constraint to wider application of this method is the limited number of trained homeopaths. PMID- 15892487 TI - Comparative efficacy of homeopathic and allopathic systems of medicine in the management of clinical mastitis of Indian dairy cows. AB - Mastitis is the major problem of dairy animals despite a number of preventive and therapeutic approaches. Treatment is costly and out of reach of farmers of developing countries like India. The treatment cost of bovine mastitis with conventional treatment has been calculated. Good results have been claimed with homeopathic treatment however, treatment costs are not available. This article reports the treatment economics of homeopathic drugs conventional drugs for the management of bovine mastitis. Ninety-six mastitic quarters (non-fibrosed 67 and fibrosed 29) were treated with a homeopathic combination medicine. Another 96 quarters with acute mastitis (non-fibrosed) treated with different antibiotics were included in the study. The animals were selected from dairy farm of the Indian Veterinary Research Institute and from private dairy farms. The overall effectiveness of homeopathic combination medicine in the treatment of acute non fibrosed mastitis was 86.6% with a mean recovery period of 7.7 days (range 3-28), and total cost of therapy as Indian Rupees 21.4 (0.39 Euros, US$ 0.47). The corresponding cure rate for the antibiotic group was 59.2% with a mean recovery period of 4.5 days (range 2-15) and an average treatment cost of Rs.149.20 (2.69 Euros, US$ 3.28). We conclude that the combination of Phytolacca, Calcarea fluorica., Silica, Belladonna, Bryonia, Arnica, Conium and Ipecacuanha (Healwell VT-6) was effective and economical in the management of mastitis in lactating dairy cows. PMID- 15892488 TI - Histamine at high dilution reduces spectral density in delta band in sleeping rats. AB - Histamine is a central neurotransmitter, it increases arousal via H1 receptors. This study examines the effect of ultra-diluted histamine on arousal through changes in the sleep pattern of Wistar rats. The spectral density in delta (0.5 2.5 Hz) band, one of the three major spectral components of the sleep electroencephalogram, was analyzed against time. Rats were randomized to receive histamine 30c (histamine 30c, 0.05 ml every 20 min during the first 2 h orally), histamine intraperitoneal pre-treatment/histamine 30c (histamine 6mg/kg i.p., followed by histamine 30c) or solvent control. The mean delta band spectral density was lower in the histamine 30c and histamine pretreatment/histamine 30c groups than the control group. Significant differences between histamine 30c and baseline during the first 2 h imply an immediate effect. These results also suggest a dynamic process in which the system spontaneously evolves between two locally stationary states according to a power law. From the time perspective, the system approaches, asymptotically, an equifinal state. PMID- 15892489 TI - Presentiment or entanglement? An alternative explanation for apparent entanglement in provings. AB - A number of authors have recently discussed the possible role entanglement in homeopathy. Walach et al have published a homeopathic proving which they interpreted as demonstrating entanglement between placebo and verum groups in a proving. The lack of a 'run-in' period was a weakness of this trial. We present further results of our proving of Belladonna which show that subjects who reported symptoms during the placebo run-in period ('presentiment provers') were more likely to report symptoms during the treatment period. This data suggests and the observations of Walach et al may be explicable by conventional mechanisms including differential reporting and constitutional type. PMID- 15892490 TI - Entangled--and tied in knots! Practical consequences of an entanglement model for homeopathic research and practice. AB - The assertion that 'local' theories of homeopathy are traditional appears to be contradicted by Hahnemann's description of the action of homeopathic medicines as 'spirit-like'. Entanglement theory prohibits the use of entangled states to convey information. Experimental proof of entanglement can only come indirectly. The implications for clinical research include that positive results will probably be found only in large series and that studies should avoid imposing a causal framework. PMID- 15892491 TI - The sound of two hands clapping: could homeopathy work locally and non-locally? AB - Homeopathy might require both local and non-local mechanisms to describe fully its mode of action. The increased prevalence of self-prescribing does not necessarily refute the possibility of non-local mechanisms. PMID- 15892492 TI - Entanglement and some heretical thoughts about homeopathy. AB - This paper proposes the emergent entanglement theory of homeopathy. This is based on the lack of evidence that choice of homeopathic medicine is important and predicts links between effectiveness of homeopathic medicines and their manufacturers. It predicts that there will be a consistent variation, in terms of outcome, between homeopaths, and between medicines made by different manufacturers, but not the specific homeopathic medicines prescribed. This theory is potentially testable. PMID- 15892493 TI - Priorities and methods for developing the evidence profile of homeopathy. Recommendations of the ECH general assembly and XVIII Symposium of GIRI. AB - To achieve scientific acceptance, homeopathy must investigate several questions: 1. The activity of very highly diluted preparations. The consensus of the meeting was that there is clear evidence of this. 2. The content of very highly diluted homeopathic preparations. More research is needed but evidence exists that a specific signal is present in homeopathic preparations. 3. A theoretical framework in which the effects of homeopathic diluted preparations can be explained. The 'Body Information Theory' is such a theory. 4. The clinical effectiveness of homeopathy. Because they avoid the placebo effect, animal studies are a priority. For human trials using Quality of Life questionnaires, studies on the activity, content and theoretical basis of homeopathic preparations were reviewed approximately 70% of cases; more in children showed improvement. Homeopathy reduced costs and allowed a better improvement in work days lost compared with conventional practice. Randomised controlled trials (RCTs) implicitly test the placebo hypothesis; RCTs have been performed and meta analyses conclude that there is clear evidence of efficacy which cannot be attributed to placebo effect. Priorities depend on the audience. More research is needed especially regarding the content of homeopathic preparations and the transmission of information. Theoretical issues are also important to avoid incorrect design of research protocols. More effort should be dedicated to veterinary research. Clinical effects analysis in humans remains important. Many other questions should be prioritised, such as the potential of homeopathy to avoid invasive procedures in children and the long-term effects of homeopathy in preventing chronic complications. PMID- 15892494 TI - Sanicula aqua: three clinical cases and a heuristic. AB - The cases presented are of three children who responded well to Sanicula aqua. I have used the exact words of the patients or their mothers. All 3 are: firstborn girls, tall and very thin with blue-green eyes. Two with recurrent otitis media with perforated tympanic membrane and smell of rotting fish, the third atopic eczema and asthma. The case analyses, materia medica of Sanicula aqua and the creation of a remedy heuristic, are discussed. PMID- 15892495 TI - 20 years ago: The British Homeopathic Journal, April 1985. PMID- 15892496 TI - Dental caries in rats. PMID- 15892497 TI - Politicians should keep their distance. PMID- 15892498 TI - Depression: what every nurse should know. PMID- 15892499 TI - Putting casualty back on its feet. PMID- 15892500 TI - The use of complementary therapy in nursing practice. AB - The use of complementary or alternative medicines (CAM) to supplement orthodox medicine is becoming more widely accepted. This article discusses why this is an expanding area of practice and outlines the risks and benefits of using CAM. PMID- 15892501 TI - Travellers' diarrhoea. PMID- 15892502 TI - Dihydrocodeine tartrate. PMID- 15892503 TI - Treatments and devices for future diabetes management. AB - This last article in this three-part series discusses the evolution of diabetes therapy and examines new treatments and delivery devices that are under development to help meet treatment and management targets. PMID- 15892504 TI - Joint primary-secondary care design of PEG care pathways. AB - The use of PEG feeding has increased over the past decade. This article describes the development of a care pathway to support patients and maintain standards of care in PEG feeding. PMID- 15892505 TI - Development of a continence assessment pathway in acute care. AB - Incontinence is defined as any loss of urine or faeces that is involuntary (Abrahams et al, 2002). It can affect anyone at any age. The prevalence of incontinence increases with age, and in women it is often associated with childbearing and the menopause. Incontinence can have devastating effects on an individual's life (Laycock et al, 2001), with both emotional and financial costs (Paddison, 2002). In the UK, conservative estimates suggest that 424 million pounds per year is spent on urinary incontinence alone (Continence Foundation, 2000). PMID- 15892506 TI - The Eton Scale: a tool for risk assessment for constipation. AB - The Eton Scale Risk Assessment for Constipation (Fig 1) will be officially launched at the Association for Continence Advice conference in Telford next week. This follows successful evaluation of the tool by nurses, who found that it was easy and simple to use. Further testing of the tool's sensitivity and reliability is currently being undertaken, and a randomised controlled trial has been planned for later in the year. PMID- 15892507 TI - SIGN guidelines on managing urinary incontinence. AB - Urinary incontinence is not directly life-threatening but it has a significant impact on the quality of life of patients, although for the vast majority relatively simple interventions can effect a cure or significant improvement (Tannenbaum et al, 2001). Many community nurses feel that their role should include assessment and first-line management of urinary incontinence but their practice is often limited by a lack of knowledge (Bignall and Getliffe, 2001). PMID- 15892508 TI - Discreet products for women with urinary incontinence. AB - It is estimated that 10 million women in the UK suffer with urinary incontinence (Hunskarr et al, 2004). Many women consider bladder weakness to be an inevitable consequence of childbirth or the menopause. Haslam (2004) reported on a telephone survey that examined whether women were aware of stress urinary incontinence, their attitude to it and the impact it had on their daily lives. In the UK, 505 women were interviewed, 206 of whom responded positively to a question about symptoms of stress urinary incontinence, giving a prevalence rate of 41 per cent. There is also evidence that most patients with urinary incontinence do not present to their doctors PMID- 15892509 TI - Management of urinary retention. AB - An inability to void urine from the bladder is described as retention of urine. It is not uncommon after surgical procedures or childbirth but is usually temporary. It also occurs as a result of obstruction or of a neurological disease such as multiple sclerosis (Getliffe and Dolman, 2003). PMID- 15892510 TI - Make the most of flexible working. PMID- 15892511 TI - Improving menus of social care. PMID- 15892512 TI - A matter of life and death. PMID- 15892513 TI - Feeling the heat. PMID- 15892514 TI - Ask the experts? Integrated falls service. PMID- 15892515 TI - Intermediate care: policy rhetoric or an effective strategy? A review of the literature. PMID- 15892516 TI - Screening for wandering among older persons with dementia. AB - Wandering is a necessary human activity for many persons with dementia. Nursing and therapy practice in the UK must adopt a more person-centred and evidence based approach to practice in this area. Risk is a potential that something might happen and remains present until it happens. Focusing on screening for wandering in those with dementia can enable nurses to use their skills to assess the person's nursing care needs in this area, prescribe care that promotes safe wandering and may begin to enable practitioners to recognise the benefits of safe wandering for the person. Proactive care planning and delivery should reduce the need for excessive crisis interventions when the person with dementia has progressed to a level of risk unacceptable to nurses. PMID- 15892517 TI - Delphi survey research and older people. AB - Delphi survey or Delphi technique has been described as an effective way of gathering information on a subject where there is currently scant evidence available. It provides an opportunity for participants to offer an expert opinion on the subject in question and helps to clarify what individuals consider to be most important about the topic under consideration. In this article the practical steps needed to construct a Delphi survey are reviewed in the context of the special circumstances associated with older adult participants. PMID- 15892518 TI - Providing the best care. PMID- 15892519 TI - Nurse international. PMID- 15892520 TI - The new online submission--author's paradise. PMID- 15892521 TI - New procedure for fully automatic occlusal surface reconstruction by means of a biogeneric tooth model. AB - The goal of this study is to present a completely new procedure for automatic occlusal surface reconstruction and to test it in several inlay situations. The starting point is the mathematical analysis of a large number of occlusal surfaces from a tooth library. Learning algorithms used in pattern recognition and image processing were adapted for the special needs of dental morphology. A biogeneric tooth model resulted which can mathematically describe a certain type of tooth only with a few parameters. This model was tested on 40 simulated inlay situations and as an example on two clinical inlay situations. The results show that fully automatic reconstruction was possible except for four cases. In a majority of the cases, no further interactive improvements would have been necessary. Because of the general, theoretically well-founded approach, fully automatic reconstruction of the occlusal surface can also be extended in the future to full crowns or reduced crown substructures (copings). PMID- 15892522 TI - Efficient e-learning with a well-founded media-didactic concept. PMID- 15892523 TI - In the name of the crown: the Cerec 3D crown-upgrade. PMID- 15892524 TI - The self-adjusting crown (SAC). AB - When previously using Cerec 3D, the occlusal surface could be designed according to morphologically functional aspects with the aid of a bite registration (static and/or dynamic). In addition, different tooth databases could be used for the design of crowns. However, the relevant design suggestion had then to be manually aligned onto the functional registration by the user. The new Cerec 3D software version R 2005 now promises the automatic adjustment of crown proposals in alignment with the corresponding functional registration. In this case, the crown design selected from the tooth databases is adjusted to the morphology of the functional registration and is therefore modified until light contact points are still present. The procedure of the "self-adjusting crown" is demonstrated by reference to a clinical case. As the name "self-adjusting crown" indicates, this automatic tool is possible only in restorations which have a single-segment preparation margin (crowns). PMID- 15892525 TI - Cerec 3D veneers with R2005--veneers a la carte. AB - Before the introduction of version R1800, only CORRELATION was available to the Cerec user for veneer design, and recourse had to be taken to the earlier Cerec 3 program for constructive veneer production. The software R1800 now also makes a tooth database available for veneers and allows copying and free insertion of any acquired tooth forms. With the R2005 version, the editing possibilities for designing veneers were further optimized; copying, mirroring and insertion of crowns were also facilitated. The situation-related selection of the CORRELATION design method, DENTAL DATABASE, and REPLICATION are described and assessed by reference to clinical situations. PMID- 15892526 TI - Margin isolation for optical impressions and adhesion. PMID- 15892527 TI - InEOS--new system component in Cerec 3D. AB - A new device for digitizing model surfaces for dental CAD/CAM applications is available with the inEOS scanner. It works according to the principle of stripe light projection. Both rotational scan mode of single prepared teeth and overview scan mode in which a complete model of the jaw can be acquired are possible. Detailed scans can be token in addition to improve the data quality. The software basis is the proven Cerec inLab 3D program. The virtually produced restorations can be milled either with the inLab milling unit or transferred to the Infinident milling center for central production. A clinical case which was treated by using the inEOS scanner is presented. PMID- 15892528 TI - Implant dentistry and the standard of care in dental education. PMID- 15892529 TI - Functional and esthetic rehabilitation in amelogenesis imperfecta with all ceramic restorations: a case report. AB - Amelogenesis imperfecta is a hereditary condition resulting in poor tooth development, severe anomalies, or complete absence of enamel. Enamel lesions may be the only characteristic of this condition or may be part of a generalized syndrome. Amelogenesis imperfecta can be characterized by enamel hypoplasia and/or hypomaturation or hypocalcification of the existing teeth. Restoration for patients with this condition should be oriented toward the functional and esthetic rehabilitation and the protection of the existing teeth. This paper presents a description of a patient's oral rehabilitation with all-ceramic veneers and crowns after extensive crown lengthening. The diagnostic procedure is also reported in detail. A microscopic examination was also performed on an extracted third molar under polarized light. PMID- 15892530 TI - Antibacterial activity of different generation dentin-bonding systems. AB - OBJECTIVES: The aim of this in vitro study was to investigate the antibacterial activities of different generation dentin-bonding systems. METHOD AND MATERIALS: The antibacterial activities of dentin-bonding systems Optibond FL Primer, Single Bond, Clearfil SE Bond Primer, and Prompt-L-Pop were evaluated against Streptococcus mutans, Streptococcus salivarius, Lactobacillus casei, and Lactobacillus acidophilus. Cervitec (1% chlorhexidine varnish) was also examined as a positive control material. Disk diffusion tests with filter paper disks (n = 10) containing a drop of each material (20 microL) were used. After incubation at 37 degrees C for 48 hours, the growth inhibition zones were measured in millimeters. RESULTS: Of all the materials tested, Optibond FL Primer, Clearfil SE Bond Primer, and Prompt-LPop showed larger growth inhibition zones than the control material for all bacterial strains. No antibacterial effect was noted for Single Bond. The results indicated that Optibond FL Primer had the strongest effect against the S mutans and L casei, and Clearfil SE Bond Primer produced the largest inhibition zone for S salivarius and L acidophilus among the test materials. CONCLUSION: The antibacterial effects observed for the tested different generation dentin-bonding systems may be related to the acidic nature of the materials. PMID- 15892531 TI - Management of hypodontia: orthodontic considerations (II). AB - Patients with congenitally missing teeth (hypodontia) present with complex problems. Meeting their treatment needs requires a joint effort between the disciplines of orthodontics and restorative dentistry. There are a number of options available to restore space generated by missing teeth. In the majority of patients orthodontic treatment will be required before this space can be restored satisfactorily. The second part of this series reviews the orthodontic considerations for various options. PMID- 15892532 TI - Clinical performance of sealants with and without a bonding agent. AB - OBJECTIVES: The objective of this study was to assess the effect of a one-bottle bonding agent on sealant success after 3, 6, 12, and 24 months. METHOD AND MATERIALS: The sample group consisted of 30 schoolchildren (8 to 10 years of age) who had all their permanent first molars sound and unsealed. Occlusal sealants were placed on first molars according to a split-mouth design. Sealant scoring occurred at 3, 6, 12, and 24 months and was performed by two clinicians blinded to the treatment group. Fisher's exact test was used to analyze the data. RESULTS: The clinically acceptable marginal integrity rates for sealants with a bonding agent after 3, 6, 12, and 24 months were 93%, 93%, 83%, and 79%, respectively. For sealants without a bonding agent, the clinically acceptable marginal integrity rates after 3, 6, 12, and 24 months were 90%, 88%, 81%, and 77%. The rates for no color change in sealants with a bonding agent after 3, 6, 12, and 24 months were 96%, 93%, 81%, and 75%, respectively. For sealants without a bonding agent, the no color change rates after 3, 6, 12, and 24 months were 93%, 93%, 79%, and 72%. After 24 months, 73% of the sample group were available for recall. The retention rates for sealants with and without a bonding agent after 24 months were 79% and 75%, respectively. The results of the statistical analysis showed that there were no differences among the sealants with and without bonding agents evaluated in relation to marginal integrity, marginal discoloration, and anatomic form. CONCLUSION: The success of a sealant is related to whether the sealant is applied under optimal conditions. The results of this study show that at the 2-year mark, the placement of a bonding agent under sealants did not significantly affect the clinical success of sealants. PMID- 15892533 TI - Water flow in high-speed handpieces. AB - OBJECTIVES: This study measured the water flow commonly used in high-speed handpieces to evaluate the water flow's influence on temperature generation. Different flow speeds were evaluated between turbines that had different numbers of cooling apertures. METHOD AND MATERIALS: Two water samples were collected from each high-speed handpiece at private practices and at the School of Dentistry at Sao Jose dos Campos. The first sample was collected at the customary flow and the second was collected with the terminal opened for maximum flow. The two samples were collected into weighed glass receptacles after 15 seconds of turbine operation. The glass receptacles were reweighed and the difference between weights was recorded to calculate the water flow in mL/min and for further statistical analysis. RESULTS: The average water flow for 137 samples was 29.48 mL/min. The flow speeds obtained were 42.38 mL/min for turbines with one coolant aperture; 34.31 mL/min for turbines with two coolant apertures; and 30.44 mL/min for turbines with three coolant apertures. There were statistical differences between turbines with one and three coolant apertures (Tukey-Kramer multiple comparisons test with P < .05). CONCLUSION: Turbine handpieces with one cooling aperture distributed more water for the burs than high-speed handpieces with more than one aperture. PMID- 15892534 TI - Clinical performance of a packable resin composite for a period of 3 years. AB - OBJECTIVE: The purpose of this clinical study was to evaluate SureFil packable resin composite for posterior restoration of permanent teeth for a period of 3 years. METHOD AND MATERIALS: Fifty-five resin composite restorations were placed in 36 patients (16 Class I and 39 Class II restorations). After cavity preparation, the enamel was etched with 34% phosphoric acid. Prime & Bond NT was applied 20 seconds to dentin and etched enamel and cured for 20 seconds. The teeth were restored in 3- to 5-mm increments. The restorations were assessed after placement, at 6 months, 1 year, 2 years, and 3 years for color stability, marginal discoloration, marginal adaptation, secondary caries, surface texture, anatomic form, and postoperative sensitivity according to Ryge's criteria. The changes in the parameters were assessed with Friedman test analysis with a Bonferroni correction at a significance level of .05. RESULTS: Forty of the monitored 47 restorations were classified as excellent after 3 years. Thirty-one restorations were graded Bravo at baseline for color match. At the 3-year assessment (n = 47) the color of the 31 restorations had not changed. Two restorations (same patient) were lost after 1 month and were scored as Charlie until the end of the study. After 3 years there were five Bravos and one Charlie with marginal discoloration, five Bravos with marginal adaptation, and three Bravos with anatomic form (P < .05). CONCLUSION: After 3 years of clinical service, SureFil packable resin composite, with a failure rate of 6%, was considered to be successful in Class I and II restorations. PMID- 15892535 TI - Effects of disinfectants on resilient denture-lining materials contaminated with Staphylococcus aureus, Streptococcus sobrinus, and Candida albicans. AB - OBJECTIVES: Resilient denture-lining materials are more susceptible to microbial adhesion than are denture base acrylic resin. As denture hygiene is essential to maintain the serviceability of the denture, disinfectant solutions have been suggested as a method for effectively cleaning resilient liners. The aim of this study was to determine the effectiveness of four different disinfectant solutions on four different resilient lining materials contaminated with Staphylococcus aureus, Streptococcus sobrinus, and Candida albicans. METHOD AND MATERIALS: Three microorganisms (Sta aureus, Str sobrinus, and C albicans), four resilient lining materials (Tempo, Immediate, Flexacryl soft, Ufi Gel P), and four disinfectant solutions (5.25% sodium hypochlorite, 2% sodium hypochlorite, 5% Deconex, and 3.5% Savlex solutions) were investigated. The four resilient lining materials were contaminated with each of the microorganisms, then the disinfection tests were applied to detect the efficacy of disinfectant solutions on resilient denture base materials in an in vitro test. The specimens (n = 5, total = 240) were soaked for 5 minutes in each solution to disinfect the microorganisms. The control specimens (one of each material; n = 5, total = 60) were soaked in distilled water for 5 minutes. As a result, a reduction in microorganism counts was determined. The Kruskal-Wallis and one-way analysis of variance tests were used to analyze for significant differences among test groups at the level of significance. RESULTS: For all microorganisms, soaking in 5.25% sodium hypochlorite reduced the number of viable adherent microorganisms significantly compared to soaking in 2% sodium hypochlorite, which led to greater reduction than soaking either 5% Deconex or 3.5% Savlex. The use of 5.25% sodium hypochlorite in all groups is statistically significant. CONCLUSION: Disinfection of four different resilient denture-lining materials with 5.25% sodium hypochlorite solution was the most effective immersion technique. To achieve an efficient disinfection, the immersion time of the other disinfectant solutions could be increased. PMID- 15892536 TI - The medical oncology patient. AB - Chemotherapy may be either an effective alternative to surgery or a valuable adjunct to surgery and/or radiotherapy in the treatment of cancer. The care of medical oncology patients is a multidisciplinary effort. Oral health care providers can expect to be called on to care for patients with cancer undergoing chemotherapy. Early, active participation in developing preventive and therapeutic strategies, in implementing the plan, and in the education and rehabilitation of patients is paramount in addressing quality of life issues. To provide timely and competent care, oral health care providers must understand the disease, its treatment, and the impact the disease and/or its treatment may have on these patients. Oral health care providers should develop and implement preventive and therapeutic strategies with the same ethical, moral, and professional standards of care as may be appropriate in the management of any other patient. PMID- 15892537 TI - Apical periodontitis of maxillary premolar with resultant inflammation of the sinus mucosa. PMID- 15892538 TI - Color vision tests for aviation: comparison of the anomaloscope and three lantern types. AB - INTRODUCTION: A comparison of the results obtained with the Nagel anomaloscope and the Holmes-Wright Type A, Spectrolux, and Beyne aviation color vision lanterns was undertaken. The Joint Aviation Requirements (JAR) specify pass/fail limits for these four secondary color vision tests and the Ishihara screening test. The results for individuals on all five tests were studied. METHODS: The color vision of 55 color-vision deficient and 24 color-vision normal subjects, mostly applicant pilots, was assessed using a battery of tests, including the Ishihara plates, the Nagel anomaloscope, and three lanterns. The testing methods and characteristics of the lanterns and anomaloscope were compared. RESULTS: Of the color-deficient applicants, only deuteranomalous trichromats passed more than one of the four secondary JAR tests, but a pass on one test did not reliably predict a pass on another test. Three out of nine protanomalous trichromats passed the Nagel anomaloscope but failed all three lantern tests. Of the normal trichromats, 12 failed the anomaloscope and 12 failed the Beyne lantern. DISCUSSION: Variability in pass/fail results can be attributed to many factors apart from loss of chromatic sensitivity. Some normal trichromats can fail both the Ishihara screening and the secondary tests. The approved secondary test varies between countries and the outcome of regulatory assessment depends on the color vision test used. Since the flight safety consequences of the current situation cannot be ignored, the development of a less variable technique for color vision assessment that is accepted internationally, allied with a better understanding of color vision requirements, is needed. PMID- 15892539 TI - Hemodynamics of graded water immersion in the baboon: +Gz protection potential. AB - INTRODUCTION: Fluid-filled anti-G suits depend on external fluid counterpressure to maintain near-normal hemodynamics during +Gz (head-to-foot inertial force). However, the relationship between external fluid level, hemodynamic events, and +Gz tolerance has not been determined. METHODS: Seven anesthetized male baboons (avg. wt. = 27.2 kg) were catheterized for measurement of left ventricular pressure (LVP), right ventricular pressure (RVP), aortic BP (ABP), central venous pressure (CVP), pulmonary artery pressure, cardiac output, and esophageal pressure. The animals were seated in a restraint chair and control data were collected, followed by six discrete steps of water immersion (WI): 1) knee; 2) hip; 3) xiphoid; 4) mid-chest; 5) neck; and 6) return to the xiphoid (data check). Each level was maintained until hemodynamic stabilization. Rectal temperature averaged 33.5 degrees C; water bath temperature was maintained at 34 degrees C. RESULTS: There was a significant (p < 0.05) increase in most hemodynamic parameters with WI to the xiphoid, mid-chest, and neck. With WI to the neck, CVP, LV, and RV end diastolic pressures increased by 8.7 mmHg, 19.6 mmHg, and 8.0 mmHg, respectively, suggesting a major passive increase in thoracic and cardiac blood volume, whereas mean ABP (MABP) increased by 18.6 mmHg and total peripheral resistance significantly decreased. Heart rate did not change significantly. CONCLUSION: The increase in MABP with WI to the neck would provide slightly more than +1 Gz relaxed protection. Additional +Gz would require pressure breathing to overcome the increased external hydrostatic pressure on the thorax. Water-filled anti-G suits filled to the xiphoid cannot provide adequate relaxed +Gz protection for current high performance aircraft without supplemental factors such as additional pressure in the suit or positive pressure breathing. PMID- 15892540 TI - Wrist vibration affects the production of finely graded forces. AB - INTRODUCTION: It is still unclear how central commands and afferent feedback interact to produce finely graded forces. To explore this question, we experimentally degraded afferent responsiveness using wrist vibration. METHODS: Subjects grasped an isometric joystick with their preferred hand and produced forces of different magnitudes and directions according to visually presented vectors. In one condition, the dorsal and palmar sides of the wrist were vibrated at 80 Hz to degrade proprioception; in another condition, the wrist was not vibrated. RESULTS: Response magnitude averaged 2.46 kg of force (kgf) without, and decreased to 1.90 kgf with vibration. The coefficient of magnitude variation was vibration-independent and averaged 0.30 kgf. The directional error of responses and its standard deviation were also vibration-independent, averaging 2.6 degrees and +/-17 degrees, respectively. Subjects' maximum voluntary force was 9.77 kgf without, and decreased to 8.83 kgf with vibration. CONCLUSION: Our findings clearly indicate that afferent feedback plays a role in the production of finely graded forces, probably by facilitating descending motor commands. PMID- 15892541 TI - Jumping in simulated and true microgravity: response to maximal efforts with three landing types. AB - BACKGROUND: Exercise is a promising countermeasure to the physiological deconditioning experienced in microgravity, but has not proven effective in eliminating the ongoing loss of bone mineral, most likely due to the lack of high impact forces and loading rates during in-flight activity. We wanted to determine lower-extremity response to high-impact jumping exercises in true and simulated microgravity and establish if 1-G force magnitudes can be achieved in a weightless environment. METHODS: Jumping experiments were performed in a ground based zero-gravity simulator (ZGS) in 1 G, and during parabolic flight with a gravity-replacement system. There were 12 subjects who participated in the study, with 4 subjects common to both conditions. Force, loading rates, jump height, and kinematics were analyzed during jumps with three distinct landings: two-footed toe-heel, one-footed toe-heel, and flat-footed. Gravity replacement loads of 45%, 60%, 75%, and 100% bodyweight were used in the ZGS; because of time constraints, these loads were limited to 60% and 75% bodyweight in parabolic flight. RESULTS: Average peak ground-reaction forces during landing ranged between 1902+/-607 and 2631+/-663 N in the ZGS and between 1683+/-807 and 2683+/-1174 N in the KC-135. No significant differences were found between the simulated and true microgravity conditions, but neither condition achieved the magnitudes found in 1 G. CONCLUSION: Data support the hypothesis that jumping exercises can impart high impact forces during weightlessness and that the custom-designed ZGS will replicate what is experienced in true microgravity. PMID- 15892542 TI - Luminous efficiency and the measurement of daytime displays, signals, and visors. AB - PURPOSE: Measurements concerning the usability or safety of optical equipment are based on assumptions regarding luminous efficiency. The current luminous efficiency functions are derived from human sensitivity experiments taken at low light levels compared with the outdoor daytime environment. The amount of error induced by extrapolating from low light level data to high light level applications is not known. We sought to determine whether standard luminous efficiency curves CIE V(lambda) and CIE Heterochromatic Brightness Matching are appropriate for measuring day-use optical equipment such as display phosphors, lasers, LEDs, and laser eye protection, which are becoming more common in aviation. METHODS: Flicker photometry and successive heterochromatic brightness matching were used to measure changes in luminance efficiency functions with increasing levels (1, 10, 100, and 1000 fL) of light adaptation. RESULTS: Luminous efficiency was found to depend on both the method and the reference intensity with which the measurements were taken. For heterochromatic brightness matching, luminous efficiency increased for longer wavelengths as reference intensity increased. Peak luminous efficiency shifted from approximately 540 nm to greater than 600 nm with increasing intensity. Peak luminous efficiency was constant for flicker photometry across all intensities, but the function narrowed slightly at 100 fL. CONCLUSION: Luminous efficiency curves measured at high reference intensities are substantially different from the standard luminous efficiency functions. Caution should be used when measuring spectrally narrow and bright sources such as lasers and LEDs with a V(lambda) corrected photometer because the measured luminance may correlate poorly with perceived brightness. PMID- 15892543 TI - Pressure breathing without a counter-pressure vest does not impair acceleration tolerance up to 9 G. AB - INTRODUCTION: This study was to determine whether safe and adequate G-protection by pressure breathing during G (PBG) could be maintained if the COMBAT EDGE counter-pressure vest were eliminated to ensure aircrew do not unnecessarily endure a possible in-flight discomfort or distraction. METHODS: Centrifuge exposures up to +9 Gz were completed by 11 subjects, including 5 F-15 aircrew, using PBG at 60 mmHg pressure with and without the counter-pressure vest. Additional G-exposures using pressures of 0, 30, and 45 mmHg were performed without the vest. RESULTS: Elimination of the COMBAT EDGE counter-pressure vest did not significantly reduce G-tolerance. During gradual onset G exposure, the mean G level reached with PBG was 8.4 G without the vest and 8.2 G with the vest. In comparison, 6.7 G was reached without PBG. Mean times at G with rapid onset G exposure were 59 and 60 s, respectively, compared with 49 s without PBG. PBG, with or without the vest, was preferred by all test subjects. PBG at 60 mmHg produced the highest G protection and was preferred by the test subjects over lesser pressures. Subjects reported no adverse effects from the use of PBG without chest counter-pressure. CONCLUSION: The use of PBG and the anti-G straining manuever (AGSM) together enhances G tolerance and comfort more than AGSM alone. Elimination of the counter-pressure vest during use of PBG does not hinder an individual's ability to reach +9 Gz or complete a short duration simulated aerial combat maneuver G exposure. Further research is needed to determine if use of PBG without chest counter-pressure increases fatigue during multiple sorties or produces other aeromedical problems in operational environments. PMID- 15892544 TI - Psychological test profiles of USAF pilots before training vs. type aircraft flown. AB - BACKGROUND: Student pilots in the USAF are selected for fighter, bomber, or airlift/tanker tracks after basic flight training. This selection needs to be accurate in order to save time and training costs. The objective of this study was to determine whether significant psychological differences exist between pilots flying different types of aircraft and whether these differences could predict who will become a fighter pilot (FP) vs. a bomber pilot (BP) or airlift/tanker pilot (AP). METHODS: Pilots who took the Multidimensional Aptitude Battery (MAB) and NEO Personality Inventory Revised (NEO-PI-R) were linked to their aircraft type using primary USAF specialty codes. The data for 2105 pilots was analyzed using MANOVA and Bonferroni post hoc analysis to evaluate for relationships between test results and airframe assignment. RESULTS: A statistically significant difference was found between FP and AP pilot means on all segments of the MAB and portions of the NEO-PI-R. The mean scores of the FP group were higher on all IQ facets of the MAB. On the NEO-PI-R, the FP group scored lower on agreeableness and higher on conscientiousness. DISCUSSION: The homogeneity of the pilot population gives the statistical difference in scores limited practical value for predicting which aircraft a pilot is best suited to fly. However, scores on these tests clearly could be a useful adjunct, along with flight training grades and personal desires, in determining a student pilot's potential for success in the multi-tasking environment of the fighter pilot. PMID- 15892545 TI - Extravehicular mobility unit training and astronaut injuries. AB - BACKGROUND: Astronaut spacewalk training can result in a variety of symptom complaints and possible injuries. This study quantified and characterized signs, symptoms, and injuries resulting from extravehicular activity spacesuit training at NASA's Neutral Buoyancy Laboratory, Johnson Space Center, Houston, TX, immersion facility. METHODS: We identified the frequency and incidence of symptoms by location, mechanisms of injury, and effective countermeasures. Recommendations were made to improve injury prevention, astronaut training, test preparation, and training hardware. At the end of each test, a questionnaire was completed documenting signs and symptoms, mechanisms of injury, and countermeasures. RESULTS: Of the 770 tests, there were 190 in which suit symptoms were reported (24.6%). There were a total of 352 reported suit symptom comments. Of those symptoms, 166 were in the hands (47.16%), 73 were in the shoulders (20.7%), and 40 were in the feet (11.4%). Others ranged from 6.0% to 0.28%, respectively, from the legs, arms, neck, trunk, groin, and head. Causal mechanisms for the hands included moisture and hard glove contacts resulting in fingernail injuries; in the shoulders, hard contact with suit components and strain mechanisms; and in the feet, hard boot contact. The severity of symptoms was highest in the shoulders, hands, and feet. CONCLUSIONS: Most signs and symptoms were mild, self-limited, of brief duration, and were well controlled by available countermeasures. Some represented the potential for significant injury with consequences affecting astronaut health and performance. Correction of extravehicular activity training-related injuries requires a multidisciplinary approach to improve prevention, medical intervention, astronaut training, test planning, and suit engineering. PMID- 15892546 TI - Modeling physiological responses to military scenarios: initial core temperature and downhill work. AB - INTRODUCTION: Previous field studies suggested that a thermoregulatory model developed by the U.S. Army Research Institute of Environmental Medicine (USARIEM) needed an adjustment of initial core temperature (Tcr) for individual variation and a metabolic (M) correction during downhill movements. This study evaluated the updated version of the model incorporating these new features using a dataset collected during U.S. Marine Corps marksmanship training at Quantico, VA. METHODS: Individual anthropometrics, physiological, and environmental time series data were obtained from five Marine men. The study focused on the marksmanship training for approximately 2 h, then 30-min marching including uphill and downhill movements in a moderately hot environment (air temperature: approximately 30 degrees C; dew point: approximately 21 degrees C). The predicted and observed heart rate (HR) and Tcr measurements were compared by root mean square deviations (RMSD). RESULTS: Overall, the current model improved predictions of physiological measures (HR RMSD = 23 bpm, Tcr RMSD = 0.46 degrees C), particularly for marching in the heat (HR RMSD = 21 bpm, Tcr RMSD = 0.32 degrees C). The model under-predicted both HR and Tcr during marksmanship training, indicating that a greater solar effect or non-thermal factors may have required higher M rates during these periods. CONCLUSIONS: Updated features of the model significantly improved physiological predictions. However, accurate M estimates are required for slow movements of subjects under heat stress, such as movements on the firing range. Such improvement should result in more accurate simulations of physiological status and better risk assessment, thereby reducing heat injuries and improving performance of deployed military personnel. PMID- 15892547 TI - Mood change and immune status of human subjects in a 10-day confinement study. AB - INTRODUCTION: Stress is a known factor that causes changes in leukocyte distribution or depression in lymphocyte proliferation. We reported previously that a 10-d confinement caused changes in immune status. Here we report the relationship between mood changes and immune parameters in the subjects confined for 10-d. METHODS: There were 10 subjects (age 20-27 yr, mean 22.8 yr) who participated in a 10-d confinement study. They were divided into 2 groups with regard to their psychological aspects and their immune parameters were then compared. Blood samples were taken once before, three times during, and once after confinement. The percentages of granulocytes, natural killer (NK) cells, and cells positive for CD69, an early activation marker, were analyzed by flow cytometry. Face scale was employed to estimate subjects' mood. RESULTS: The group that showed an increase in mood scale toward the end of the confinement showed changes in all immune parameters. In contrast, less marked changes were seen in the group that showed no mood changes throughout the experiment. DISCUSSION: These results indicate that the observed immune changes were related to the mood changes, and that mood change seems to be one of the causes of the immunological changes seen in confined environments, such as in space stations or submarines. The percentages of NK cells, granulocytes, and CD69 expression may be useful criteria for detecting immunological deterioration caused by stress, or for selecting astronauts who are immunologically stable against the challenge of confinement stress. PMID- 15892548 TI - Fatigue assessment in the field: validation of a hand-held electronic psychomotor vigilance task. AB - INTRODUCTION: In recent years, there has been an increasing need for a reliable and practical tool for assessing fatigue-related impairment in the field. This study investigated the sensitivity of one potential tool, a 5-min version of the psychomotor vigilance task (PVT) specifically designed for use on personal digital assistants (PDA), to 28 h of sustained wakefulness. METHODS: There were 15 participants who slept in the laboratory overnight then remained awake from 08:00 (Day 1) to 12:00 (Day 2). During every second hour, they completed a 10-min PVT, a sustained attention task that is sensitive to the effects of sleep loss and fatigue, and a 5-min PDA-PVT. RESULTS: While performance on both tasks significantly varied as a function of hours of wakefulness, responses on the PDA PVT were typically slower than on the PVT. When performance scores were standardized, the negative impact of increasing hours of wakefulness on performance on the 5-min PDA-PVT and 10-min PVT did not significantly differ. DISCUSSION: The findings suggest that the 5-min PDA-PVT may provide a reasonable substitute for the 10-min PVT, particularly in circumstances where a shorter test is required and/or the standard PVT is not as practical. PMID- 15892549 TI - Bubble incidence after staged decompression from 50 or 60 msw: effect of adding deep stops. AB - OBJECTIVES: The French Navy uses the Marine Nationale 90 (MN90) decompression tables for air dives as deep as 60 msw. The resulting incidence of decompression sickness (DCS) for deep dives (45-60 msw) is one case per 3000 dives. METHODS: Three protocols with experimental ascent profiles (EAPs) were tested in the wet compartment of a hyperbaric chamber. For each protocol, eight subjects dove to 50 or 60 msw and ascended according to the standard MN90 table or an EAP. Precordial bubbles were monitored with Doppler sensors at 30-min intervals after surfacing. Protocol I went to 60 msw and used deep stops beginning at 27 msw. Protocol II was a repetitive dive to 50 msw with a 3-h surface interval; the EAP made the first deep stop at 18 msw. Protocol III again went to 60 msw, but the EAP used a single, shorter deep stop at 25 msw. RESULTS: For Protocol I, all divers developed bubbles at Spencer grade 2-3 and still had bubbles 120 min after surfacing; there was no statistical difference between bubbling for the MN90 and EAP, but one diver presented a case of DCS after the EAP. For Protocol II, the EAP produced severe bubbling for the eight divers. Those findings led to stopping the EAPs with the longer deep stops used in Protocols I and II. Protocol III again showed no difference between the standard and modified profiles. DISCUSSION: The addition of deep stops requires careful consideration. Two of our EAPs made no difference and one produced increased bubbling. PMID- 15892550 TI - Centrifuge training program with "push-pull" elements. AB - INTRODUCTION: Pilots of fighter aircraft are often exposed to maneuvers that produce negative acceleration (-Gz) immediately followed by positive acceleration (+Gz). This sequence has been found to reduce tolerance to +Gz, a phenomenon known as the "push-pull" effect. We devised a centrifuge training program to demonstrate this phenomenon to pilots. METHODS: The centrifuge of the Military Institute of Aviation Medicine in Warsaw, Poland, was modified in 1996 to allow active positioning of the gondola during rotation. Head-down position of -6 degrees to -40 degrees were used to produce relative -Gz (r-Gz) in a range down to 0.2. As a side effect, this produces Gy acceleration between -1.3 Gy and -1.6 Gy. Pilots completed normal centrifuge training, including a relaxed, gradual onset run and three rapid-onset runs. They were then exposed to a profile that included a series of push-pull exposures where r-Gz was followed by +Gz with stepwise increases in the latter from +2.5 to +5 Gz. The final profile was a simulated aerial combat maneuver with push-pull elements. RESULTS: The trainees expressed surprise at the push-pull effect, which forced them to begin an anti-G straining maneuver at lower levels than normal. They complained about the presence of the Gy, which rarely occurs in aircraft. DISCUSSION: This type of profile appears useful for training pilots about the push-pull phenomenon. After collection of additional data, the profiles may be refined. PMID- 15892551 TI - G-induced visual and cognitive disturbances in a survey of 65 operational fighter pilots. AB - INTRODUCTION: Only one previous study has assessed almost loss of consciousness (A-LOC) in operational fighter pilots, reporting an incidence rate of 14%. Research also indicates that 8-13% of pilots have experienced G-induced loss of consciousness (G-LOC). A-LOC can be as insidious as G-LOC due to the associated altered state of awareness and relative incapacitation time, making it a significant risk factor in the high +Gz environment. Royal Australian Air Force (RAAF) pilots currently fly the F/A-18 and Hawk 127, producing +Gz accelerations up to +7.5 Gz, which places these pilots at risk of both A-LOC and G-LOC. METHODS: A survey was administered to 100 active RAAF fighter pilots requesting information on G-induced visual and cognitive disturbances, A-LOC symptoms, and G LOC. Details regarding type of aircraft, flying maneuvers performed and mission outcome were also sought. RESULTS: There were 65 RAAF fighter pilots who completed the survey (age 20-53 yr, height 168-193 cm, weight 64-110 kg, jet hours 30-5700 h). Of these pilots, 98% indicated they had experienced at least one visual or cognitive disturbance in the high G environment: gray-out 98%; black-out 29%; and A-LOC symptoms 52%, including abnormal sensation in limbs, disorientation, and confusion. There were 9% who indicated they had experienced G LOC (50% were the pilot flying the aircraft). DISCUSSION: These findings indicate that RAAF fighter pilots are experiencing a similar rate of visual disturbances and G-LOC when compared with other air forces. However, RAAF pilots reported a much higher incidence of A-LOC compared with the only other study of operational fighter pilots. PMID- 15892552 TI - Human factors analysis and classification system applied to civil aircraft accidents in India. AB - INTRODUCTION: The Human Factors Analysis and Classification System (HFACS) has gained wide acceptance as a tool to classify human factors in aircraft accidents and incidents. This study on application of HFACS to civil aircraft accident reports at Directorate General Civil of Aviation (DGCA), India, was conducted to ascertain the practicability of applying HFACS to existing investigation reports and to analyze the trends of human factor causes of civil aircraft accidents. METHOD: Accident investigation reports held at DGCA, New Delhi, for the period 1990--99 were scrutinized. In all, 83 accidents occurred during this period, of which 48 accident reports were evaluated in this study. RESULTS: One or more human factors contributed to 37 of the 48 (77.1%) accidents. The commonest unsafe act was 'skill based errors' followed by 'decision errors.' Violations of laid down rules were contributory in 16 cases (33.3%). 'Preconditions for unsafe acts' were seen in 23 of the 48 cases (47.9%). A fairly large number (52.1%) had 'organizational influences' contributing to the accident. DISCUSSION: These results are in consonance with larger studies of accidents in the U.S. Navy and general aviation. Such a high percentage of 'organizational influences' has not been reported in other studies. This is a healthy sign for Indian civil aviation, provided effective remedial action for the same is undertaken. PMID- 15892553 TI - Basic life support in microgravity: evaluation of a novel method during parabolic flight. AB - BACKGROUND: If a cardiac arrest occurs in microgravity, the aim of current emergency procedures is to treat the patient using a medical restraint system within 2 min. The patient may require treatment while medical equipment is being deployed. The capability for one person, unaided, to successfully perform cardiopulmonary resuscitation (CPR) is, therefore, of paramount importance. A new technique has been developed whereby the practitioner encircles the thorax of the patient with his/her legs to restrain the patient to allow CPR to be performed in microgravity. METHOD: Two investigators performed both this method (during parabolic microgravity) and traditional CPR (at +1 Gz) on an instrumented CPR mannequin. The mannequin was modified to ensure accurate chest compression and ventilation measurements during microgravity. RESULTS: The mean (+/-SE) depth and rate of chest compression were 44.0+/-4.99 mm and 68.3+/-17.0 compressions x min( 1) respectively. Although the mean microgravity rate of compression proved significantly less (p < 0.05) than the +1 Gz mean (97.1+/-3.4 compressions x min( 1)), chest compression depth did not differ (p > 0.05) from +1 Gz measures (43.6+/-0.59 mm). The mean (+/-SE) microgravity tidal volume (VT) was 491+/-50.4 ml, which also did not differ (p > 0.05) from +1 Gz values (507.6+/-11.5 ml). DISCUSSION: Although difficulties in performing this method during parabolic flight primarily affected compression rate, it may be possible to conduct basic life support using this technique in any microgravity environment. PMID- 15892554 TI - President's page. PMID- 15892555 TI - Cervical spine injury prediction model. PMID- 15892557 TI - Mentorship--we need you--every one of you! PMID- 15892556 TI - This month in Aerospace Medicine history--May 2005. PMID- 15892558 TI - Flying high with Dr. Eleanor C. (Connie) Mariano. Interview by Dale Orford. PMID- 15892560 TI - Association of Biomolecular Resource Facilities 2005 meeting. Biomolecular technologies: discovery to hypothesis. PMID- 15892561 TI - Electrophoresis in 2004: the Annual Meeting of the American Electrophoresis Society. PMID- 15892562 TI - Macromolecular crystallization with microfluidic free-interface diffusion. AB - Fluidigm Corp. released the Topaz 1.96 and 4.96 crystallization chips in the fall of 2004. Topaz 1.96 and 4.96 are the latest evolution of Fluidigm's microfluidics crystallization technologies that enable ultra-low-volume rapid screening for macromolecular crystallization. Topaz 1.96 and 4.96 are similar to each other but represent a major redesign of the Topaz system and have substantially improved ease of automation and ease of use, improved efficiency and even further reduced the amount of material needed. With the release of the new Topaz system, Fluidigm continues to set the standard in low-volume crystallization screening, which is having an increasing impact in the field of structural genomics and more generally in structural biology. It is likely that further optimization and increased utility of the Topaz crystallization system will emerge. It is also probable that further innovation and the emergence of competing technologies will be seen. PMID- 15892563 TI - A streamlined approach to high-throughput proteomics. AB - Proteomics has rapidly become an important tool for life science research, allowing the integrated analysis of global protein expression from a single experiment. To accommodate the complexity and dynamic nature of any proteome, researchers must use a combination of disparate protein biochemistry techniques, often a highly involved and time-consuming process. Whilst highly sophisticated, individual technologies for each step in studying a proteome are available, true high-throughput proteomics that provides a high degree of reproducibility and sensitivity has been difficult to achieve. The development of high-throughput proteomic platforms, encompassing all aspects of proteome analysis and integrated with genomics and bioinformatics technology, therefore represents a crucial step for the advancement of proteomics research. ProteomIQ (Proteome Systems) is the first fully integrated, start-to-finish proteomics platform to enter the market. Sample preparation and tracking, centralized data acquisition and instrument control, and direct interfacing with genomics and bioinformatics databases are combined into a single suite of integrated hardware and software tools, facilitating high reproducibility and rapid turnaround times. This review will highlight some features of ProteomIQ, with particular emphasis on the analysis of proteins separated by 2D polyacrylamide gel electrophoresis. PMID- 15892564 TI - Host-pathogen interactions: a proteomic view. AB - Host-pathogen interactions reflect the balance of host defenses and pathogen virulence mechanisms. Advances in proteomic technologies now afford opportunities to compare protein content between complex biologic systems ranging from cells to animals and clinical samples. Thus, it is now possible to characterize host pathogen interactions from a global proteomic view. Most reports to date focus on cataloging protein content of pathogens and identifying virulence-associated proteins or proteomic alterations in host response. A more in-depth understanding of host-pathogen interactions has the potential to improve our mechanistic understanding of pathogenicity and virulence, thereby defining novel therapeutic and vaccine targets. In addition, proteomic characterization of the host response can provide pathogen-specific host biomarkers for rapid pathogen detection and characterization, as well as for early and specific detection of infectious diseases. A review of host-pathogen interactions focusing on proteomic analyses of both pathogen and host will be presented. Relevant genomic studies and host model systems will be also be discussed. PMID- 15892565 TI - Proteomics for biodefense applications: progress and opportunities. AB - The increasing threat of bioterrorism and continued emergence of new infectious diseases has driven a major resurgence in biomedical research efforts to develop improved treatments, diagnostics and vaccines, as well as increase the fundamental understanding of the host immune response to infectious agents. The availability of multiple mass spectrometry platforms combined with multidimensional separation technologies and microbial genomic databases provides an unprecedented opportunity to develop these much needed resources. An overview of current proteomic strategies applied to microbes and viruses considered potential bioterrorism agents is presented. The emerging area of immunoproteomics as applied to the development of new vaccine targets is also summarized. These powerful research approaches can generate a multitude of potential new protein targets; however, translating these proteomic discoveries to useful counter bioterrorism products will require large collaborative research efforts across multiple basic science and clinical disciplines. A translational proteomic research paradigm illustrating this approach using influenza virus as an example is discussed. PMID- 15892566 TI - Proteomic approaches for studying chemoresistance in cancer. AB - The role of various proteins involved in drug resistance in tumor cells is discussed in this review. Two types of studies are covered: those performed in the preproteomics era and those carried out with modern proteomic tools, namely 2D (electrophoretic) maps and 2D chromatography. In the preproteomic studies, one protein had generally been held responsible for a given chemoresistance. However, analysis via proteomic tools may reveal entire sets of proteins that are up- or downregulated (or switched on/off) in chemoresistant tumor cell lines compared with parental tumor lines. Therefore, it appears more realistic to expect that exposure of cells to drugs results in the activation of different mechanisms of resistance. Such investigations have led to the broadly shared opinion that exposure of cells to drugs results in the activation of different mechanisms of resistance, and that a specific drug-resistant phenotype consists of several molecular mechanisms that are simultaneously active. The proteomic papers reviewed clearly support the hypothesis that many metabolic pathways are affected during the resistance process. Although the modulation of expression levels of such proteins is not clear proof of their role in drug resistance per se, at least some of the themes are very likely to be involved in the resistance phenotype, and thus may be potential targets for new drugs. It is hoped that this review will bring new insight in this field and will stimulate novel and deeper searches with proteomic tools (including prefractionation of subcellular organelles, such as nuclei, to bring to the fore low-abundance proteins that might be responsible for the onset of drug resistance). PMID- 15892567 TI - Proteomics in cancer vaccine development. AB - Proteomics is a new scientific field aimed at the large-scale characterization of the protein constituents of biologic systems. It facilitates comparisons between different protein preparations by searching for minute differences in their protein expression repertoires and the patterns of their post-translational modifications. These attributes make proteomics perfectly suited for searching for proteins and peptides expressed exclusively or preferentially in cancer cells as candidates for cancer vaccines. The main proteomics technologies include 2D polyacrylamide gel electrophoresis, multidimensional high-performance liquid chromatography, mass spectrometry and protein arrays. Proteomics technologies used to analyze cancer culture cells, fresh tumor specimens, human leukocyte antigen peptides, serum and serum antibodies (serologic proteomics) have successfully identified tumor markers. Turning the potential vaccine candidates identified by proteomics technologies into clinical treatments awaits demonstration. PMID- 15892568 TI - Recent advances in 2D electrophoresis: an array of possibilities. AB - 2D electrophoresis is currently the most widespread technique used for performing functional proteomics (i.e., the large-scale analysis of alterations in protein expression levels). Nevertheless, several limitations inherent to this technology have restricted the full potential of this protein differential display methodology for years. This has even led to the abandonment of 2D electrophoresis by several groups that switched to performing gel-free functional proteomics analyses based on liquid chromatography and mass spectrometry. Meanwhile, important recent advances in 2D electrophoresis, such as the introduction of fluorescent 2D difference gel electrophoresis and numerous protein prefractionation techniques, have thoroughly modernized 2D electrophoresis, making it again one of the preferred methods for the analysis of protein expression differences in many laboratories. PMID- 15892569 TI - Shotgun lipidomics: multidimensional MS analysis of cellular lipidomes. AB - Shotgun lipidomics, comprised of intrasource separation, multidimensional mass spectrometry and computer-assisted array analysis, is an emerging powerful technique in lipidomics. Through effective intrasource separation of predetermined groups of lipid classes based on their intrinsic electrical propensities, analyses of lipids from crude extracts of biologic samples can be directly and routinely performed. Appropriate multidimensional array analysis of lipid pseudomolecular ions and fragments can be performed leading to the identification and quantitation of targeted lipid molecular species. Since most biologic lipids are linear combinations of aliphatic chains, backbones and head groups, a rich repertoire of multiple lipid building blocks present in discrete combinations represent experimental observables that can be computer reconstructed in conjunction with their pseudomolecular ions to directly determine the lipid molecular structures from a lipid extract. Through this approach, dramatic increases in the accessible dynamic range for ratiometric quantitation and discrimination of isobaric molecular species can be achieved without any prior column chromatography or operator-dependent supervision. At its current state of development, shotgun lipidomics can analyze over 20 lipid classes, hundreds of lipid molecular species and more than 95% of the mass content of a cellular lipidome. Thus, understanding the biochemical mechanisms underlying lipid-mediated disease states will be greatly facilitated by the power of shotgun lipidomics. PMID- 15892570 TI - Does O-GlcNAc play a role in neurodegenerative diseases? AB - There are several lines of evidence that the modification of proteins by cytosolic- and nuclear-specific O-linked N-acetylglucosamine (O-GlcNAc) glycosylation is closely related to neuropathologies, particularly Alzheimer's disease. Several neuronal proteins have been identified as being modified with O GlcNAc; these proteins could form part of the inclusion bodies found, for example, in the most frequently observed neurologic disorder (i.e., Alzheimer's disease; Tau protein and beta-amyloid peptide are the well known aggregated proteins). O-GlcNAc proteins are also implicated in synaptosomal transport (e.g., synapsins and clathrin-assembly proteins). Inclusion bodies are partly characterized by a deficiency in the ubiquitin-proteasome system, avoiding the degradation of aggregated proteins. From this perspective, it appears interesting that substrate proteins could be protected against proteasomal degradation by being covalently modified with single N-acetylglucosamine on serine or threonine, and that the proteasome itself is modified and regulated by O-GlcNAc (in this case the turnover of neuronal proteins correlates with extracellular glucose). Interestingly, glucose uptake and metabolism are impaired in neuronal disorders, and this phenomenon is linked to increased phosphorylation. In view of the existence of the dynamic interplay between O-GlcNAc and phosphorylation, it is tempting to draw a parallel between the use of glucose, O-GlcNAc glycosylation and phosphorylation. Lastly, the two enzymes responsible for O-GlcNAc dynamism (i.e., O-GlcNAc transferase and glucosaminidase) are both enriched in the brain and genes that encode the two enzymes are located in two regions that are found to be frequently mutated in neurologic disorders. The data presented in this review strongly suggest that O-GlcNAc could play an active role in neurodegenerative diseases. PMID- 15892572 TI - The physiological chemistry and biological activity of nitroxyl (HNO): the neglected, misunderstood, and enigmatic nitrogen oxide. PMID- 15892573 TI - Characterization of a structurally intact in situ lung model and comparison of naphthalene protein adducts generated in this model vs lung microsomes. AB - Airway epithelial cells are a susceptible site for injury by ambient air toxicants such as naphthalene that undergo P450-dependent metabolic activation. The metabolism of naphthalene in Clara cells to reactive intermediates that bind covalently to proteins correlates with cell toxicity. Although several proteins adducted by reactive naphthalene metabolites were identified in microsomal incubations, new methods that maintain the structural integrity of the lung are needed to examine protein targets. Therefore, we developed a method that involves inflation of the lungs via the trachea with medium containing (14)C-naphthalene followed by incubation in situ. The viability of this preparation is supported by maintenance of glutathione levels, rates of naphthalene metabolism, and exclusion of ethidium homodimer-1 from airway epithelium. Following in situ incubation, the levels of adduct per milligram of protein were measured in proteins obtained from bronchoalveolar lavage, epithelial cells, and remaining lung. The levels of adducted proteins obtained in lavage and epithelial cells were similar and were 20-fold higher than those in residual lung tissue. (14)C-Labeled adducted proteins were identified by matrix-assisted laser desorption ionization-time-of flight (MALDI-TOF) mass spectrometry (MS) and quadrupole-TOF MS/MS. Major adducted proteins include cytoskeletal proteins, proteins involved in folding and translocation, ATP synthase, extracellular proteins, redox proteins, and selenium binding proteins. We conclude that in situ incubation maintains structural integrity of the lung while allowing examination of reactive intermediate activation and interaction with target cell proteins of the lung. The proteins adducted and identified from in situ incubations were not the same proteins identified from microsomal incubations. PMID- 15892574 TI - Isodomoic acid C, an unusual amnesic shellfish poisoning toxin from Pseudo nitzschia australis. AB - An unusual isomer of domoic acid (1), isodomoic acid C (2), has been found in New Zealand shellfish contaminated by amnesic shellfish poisoning (ASP) toxins and was shown to be produced by a local strain of the pennate diatom Pseudo-nitzschia australis. A bulk culture of this strain was used to isolate 2. The structure was determined from spectroscopic data and was shown to correspond to that of 2 from a Japanese red seaweed, the only other reported occurrence of this compound. The affinity of 2 for GluR6 glutamate receptors was 240-fold lower than for 1, indicating low neurotoxic potential. PMID- 15892575 TI - Aldehydes in cigarette smoke react with the lipid peroxidation product malonaldehyde to form fluorescent protein adducts on lysines. AB - Cigarette smoke is a risk factor for the development of several diseases, but the exact mechanism responsible has not been well-characterized. Because modification, or adducting, of biomolecules is thought to mediate the toxic effects observed from exposure to a wide variety of harmful chemicals, this study investigated the ability of cigarette smoke to produce specific adducts on a peptide to gain insight into the likely effect on cellular proteins. We describe the modification of the epsilon-amino group of lysine contained in a test peptide with stable fluorescent adducts derived from monofunctional aldehydes occurring in cigarette smoke and malonaldehyde, a product of lipid peroxidation. Utilizing high-performance liquid chromatography, fluorescent measurements, and matrix assisted laser desorption/ionization time-of-flight mass spectroscopy, the 1,4 dihydropyridine-3,5-dicarbaldehyde and 4-methyl-1,4-dihydropyridine-3,5 dicarbaldehyde derivatives of lysine were identified as products of exposure to cigarette smoke extract and malonaldehyde. These data suggest that cigarette smoke may promote the modification of proteins, like those associated with oxidized low-density lipoprotein, and may contribute to smoking-related disease. PMID- 15892576 TI - Beauvericin activates Ca2+-activated Cl- currents and induces cell deaths in Xenopus oocytes via influx of extracellular Ca2+. AB - Beauvericin is a mycotoxin that infects a wide variety of cereal grains. The toxicological importance of beauvericin is implicated by its cytotoxicity in animal and human cells, which has been suggested to result from an increase in intracellular Ca(2+) concentration ([Ca(2+)](i)). Despite the fact that beauvericin may activate extracellular Ca(2+) influx, beauvericin-induced cell deaths has been suggested to be exclusively due to Ca(2+) release from internal Ca(2+) stores. We endeavored to elucidate the mechanism of beauvericin-induced [Ca(2+)](i) increase by studying the effects of beauvericin in Xenopus oocytes. By applying a -140-mV prepulse prior to a series of test pulses, we found that beauvericin induced small inward currents at -140 mV, followed by outwardly rectifying currents that displayed an apparent reversal potential close to the expected equilibrium potential of Cl(-). Both the inward and outward currents induced by beauvericin were blocked by niflumic acid, a specific blocker for Ca(2+)-activated Cl(-) currents (I(Cl,Ca)). Removal of extracellular Ca(2+), as well as perfusion of lanthanide, abrogated beauvericin-induced currents. Beauvericin also displayed prominent cytotoxic effects in Xenopus oocytes in a dose-dependent manner. In the absence of extracellular Ca(2+), cytotoxicity induced by 10 and 30 microM, but not 50 microM, of beauvericin was significantly diminished. Our results are consistent with the idea that beauvericin induces extracellular Ca(2+) influx, which in turn activates I(Cl,Ca) and contributes to beauvericin-induced cell deaths in Xenopus oocytes. PMID- 15892577 TI - Reductive activation of Cr(Vi) by nitric oxide synthase. AB - Chromium(VI) is a recognized toxicant whose effects have been linked to its reduction to lower oxidation states. Although Cr(VI) is reduced by several systems, it is anticipated that its reduction by nitric oxide synthase (NOS) could have significant effects in endothelial and brain cells that express high constitutive levels of the enzyme. This possibility was examined by electron paramagnetic resonance that showed the formation of a stable Cr(V) species from NOS/Cr(VI). The formation of Cr(V) was calcium/calmodulin-independent indicating that Cr(VI) to Cr(V) reduction occurs at the flavin-containing domain of NOS. Accordingly, Cr(VI) reduction by the reductase domain of NOS and the chimera protein cytochrome-P450-reductase+tail-nNOS also generated Cr(V). Activation of tetrahydrobiopterin (BH(4))-free NOS with calcium/calmodulin diminished Cr(V) steady-state levels while increasing superoxide formation. Since SOD restored Cr(V) to control levels, this result was taken as evidence for a reaction between Cr(V) and superoxide. Supplementation of NOS with BH(4) cofactor not only failed to increase Cr(V) yields but generated superoxide and hydroxyl radical. Since the holoenzyme does not generate superoxide, this reaction indicated that Cr(V) mediates the oxidation of BH(4)-bound to the enzyme. In the presence of L arginine, however, Cr(VI) neither enhances superoxide release nor inhibits NO formation from fully active NOS. This suggests that L-arginine protects BH(4) from Cr(V)-mediated oxidation. While Cr(V) was inactive toward NO, spin trapping experiments with 5-tert-butoxycarbonyl 5-methyl-1-pyrroline N-oxide and oxygen consumption measurements showed that Cr(V) reacts with superoxide by a one electron-transfer mechanism to generate oxygen and Cr(IV). Thus, reduction of Cr(VI) to Cr(V) by NOS occurs in resting and fully active states. It is likely that the reaction between Cr(V) and superoxide influences the cytotoxic mechanisms of Cr(VI) in cells. PMID- 15892578 TI - Chemistry-toxicity relationships for the effects of di- and trihydroxybenzenes to Tetrahymena pyriformis. AB - This paper presents a mechanistic analysis of aquatic toxicity data, quantified as pIGC(50) assessed in the 40 h Tetrahymena pyriformis population growth impairment assay, for 40 polyhydroxybenzene derivatives. The toxicity trends of these phenolic compounds have been shown to be consistent with mechanistic organic chemistry principles. Thus, it is shown that the compounds can be grouped into two chemical mechanism of action domains, according to whether they can be oxidized to electrophilic quinones or quinone methides. Compounds in which the hydroxy groups are oriented meta, but not ortho or para, to one another cannot be oxidized to electrophilic quinones or quinone methides and act as polar narcotics. Their toxicities are found to be well-correlated with hydrophobicity (modeled by log D): pIGC(50) = 0.83 (+/-0.04) log D - 1.27 (+/-0.09): n = 10, r(2) (adj) = 0.981, q(2) = 0.974, s = 0.15, and F = 460. Compounds with hydroxy groups oriented ortho or para to one another are more toxic than predicted by this equation, and the toxicity trends within this group of compounds are rationalized in terms of the electrophilic chemistry of their oxidation products. A quantitative correlation is demonstrated between toxicity and electrophilicity of the oxidation products, as modeled by the activation energy index (AEI), a new molecular orbital parameter derived from the computed highest occupied molecular orbital (HOMO) and HOMO-1 orbital energies of the electrophiles and the intermediates for Michael addition of n-butylamine: pIGC(50) (adj) = -0.49 (+/ 0.06) AEI + 6.85 (+/-0.69): n = 18, r(2) (adj) = 0.810, q(2) = 0.774, s = 0.24, and F = 73. Outliers to these quantitative structure-activity relationships (QSARs) are easily rationalized in terms of their chemistry (tetrabromocatechol, 4,6-dinitro-1,2,3-trihydroxybenzene, and 2,3,4-trihydroxybenzophenone) or in a demonstrable deficiency in the descriptor (the methyl-substituted hydroquinones, for which the AEI parameter as defined here fails to model the electron donation effects of the methyl groups). The AEI parameter is a mechanism-based molecular orbital parameter new to QSAR and, on the basis of the present findings, it shows promise for further applications. However, some deficiencies have been identified with it, particularly with regard to modeling the electronic effects of methyl (and presumably other alkyl) groups, and there is scope to refine the concept so as to deal with these deficiencies. PMID- 15892579 TI - Bioactivation of 4-ipomeanol by CYP4B1: adduct characterization and evidence for an enedial intermediate. AB - 4-Ipomeanol (IPO) is a pneumotoxin that is bioactivated to a reactive intermediate that binds to DNA and other cellular macromolecules. Despite over 30 years of research in this area, detailed structural information on the nature of the IPO reactive intermediate is still lacking. In the present study, we reacted IPO with rabbit CYP4B1 in the presence of exogenous nucleophiles and analyzed the products by liquid chromatography/electrospray ionization-mass spectrometry. Coincubation of IPO and rabbit CYP4B1 with glutathione gave rise to multiple products due likely to the presence of both sulfur and nitrogen nucleophiles in the same trapping molecule. Reaction mixtures containing equimolar N-acetyl cysteine (NAC) and N-acetyl lysine (NAL) provided a major NADPH- and CYP4B1 dependent product. A combination of high-resolution mass spectrometry and two dimensional NMR analysis following large-scale isolation of the biologically derived material provided evidence for an N-substituted cysteinyl pyrrole derivative of IPO, analogous to that characterized previously in model chemical studies conducted with cis-2-butene-1,4-dial. Purified native rabbit lung CYP4B1 and purified recombinant rabbit CYP4B1 produced the trapped NAC/NAL-IPO pyrrole adduct at rates of 600-700 nmol/nmol P450/30 min. A panel of 14 commercially available recombinant human CYPs was also studied, and substantial rates of IPO bioactivation (>100 nmol/nmol/30 min) were observed with CYP1A2, CYP2C19, CYP2D6, and CYP3A4. These studies provide evidence for the formation of an enedial reactive intermediate during CYP-mediated IPO bioactivation, identify multiple human liver P450s capable of IPO bioactivation, and demonstrate that the same reactive intermediate is formed by both rabbit CYP4B1 and human P450s. PMID- 15892580 TI - Identification of 4-S-Cysteinyltetrodotoxin from the liver of the puffer fish, Fugu pardalis, and formation of thiol adducts of tetrodotoxin from 4,9 anhydrotetrodotoxin. AB - The metabolic pathway of tetrodotoxin (TTX), a powerful and specific voltage gated sodium channel blocker, has not been well-clarified either in TTX-poisoned patients or in puffer fish. 4-S-CysteinylTTX (4-CysTTX) was isolated from the liver of the puffer fish, Fugu pardalis, as the first adduct of TTX with thiol. The structure was fully characterized by electrospray ionization-MS and two dimensional NMR spectroscopy. The configuration of Cys in this compound was confirmed to be S (L-Cys) by application of the Marfey's method to cystine obtained from 4-CysTTX by iodine oxidation. We also found that 4-CysTTX was derived from 4,9-anhydroTTX by incubation with a large excess of Cys in aqueous buffer (pH 8.0) for 90 min at 40 degrees C in 33% yield by HPLC. GSH also reacted with 4,9-anhydroTTX to form 4-S-glutathionylTTX (4-GSTTX) in 39% yield under the same conditions, whereas TTX scarcely reacted with Cys and GSH. These reactions were strictly pH-dependent, giving the highest yield at pH 8.0. 4-GSTTX was converted to 4,9-anhydroTTX in 0.8 M sodium phosphate buffer (pH 8.0) at 25 degrees C. Its half-life was approximately 4 h. The minimum lethal doses of 4 CysTTX and 4-GSTTX to mice by ip injection were more than 140 and 860 microg/kg (n = 2), which were 14- and 86-fold larger than the LD(50) of TTX, respectively. 4-GSTTX was hydrolyzed to 4-CysTTX by gamma-glutamyltranspeptidase (Sigma, catalog no. G9270), which was supposed to contain cysteinylglycine dipeptidase. We also examined the effect of Cys or GSH coinjection (ip) with TTX to mice for detoxification of TTX and concluded that these coinjections did not reduce the toxicity of TTX. PMID- 15892581 TI - The reactive D-glucopyranose moiety of streptozotocin is responsible for activation of macrophages and subsequent stimulation of CD8+ T cells. AB - The antitumor drug streptozotocin (STZ) is commonly used as a diabetogenic compound in animal models. At relatively low doses, STZ-induced beta cell destruction is associated with Th1-driven type 1 immune reactions, including macrophages (MPhi) and IFN-gamma-producing CD8(+) T cells. STZ induces similar Th1-dependent effects in the popliteal lymph node assay (PLNA), and because this assay allows straightforward examination of early immunostimulating processes, the PLNA was used to further examine the importance of MPhi and structural properties of STZ in relation to the induction of type 1 immune responses. Results show that elimination of MPhi with clodronate-containing liposomes prior to exposure to STZ prevents the occurrence of some (CD8(+) T cell activation, IFN gamma production, and tissue destruction) but not all (IgG2a formation) type 1 immune responses. It appeared that stimulation of MPhi depends on the d glucopyranose moiety of STZ, as well as on the intact reactive N-methyl-N nitrosourea (MNU) moiety. However, the MNU moiety suffices to induce IgG2a formation. In addition, STZ-derived nitric oxide may have modulating effects on the elicitation of STZ-induced immune responses. Present results support the idea that MPhi activation is indispensable for the STZ-induced tissue destructive type 1 responses and that various STZ-induced type 1 immune responses are differently regulated. PMID- 15892582 TI - Mechanistic studies on the metabolic scission of thiazolidinedione derivatives to acyclic thiols. AB - Thiazolidinedione (TZD) derivatives have been reported to undergo metabolic activation of the TZD ring to produce reactive intermediates. In the case of troglitazone, it was proposed that a P450-mediated S-oxidation leads to TZD ring scission and the formation of a sulfenic acid intermediate, which may be trapped as a GSH conjugate. In the present study, we employed a model compound {denoted MRL-A, (+/-)-5-[(2,4-dioxothiazolidin-5-yl)methyl]-2-methoxy-N-[[(4 trifluoromethoxy)phenyl]methyl]benzamide} to investigate the mechanism of TZD ring scission. When MRL-A was incubated with monkey liver microsomes (or recombinant P450 3A4 and NADPH-P450 reductase) in the presence of NADPH and oxygen, the major products of TZD ring scission were the free thiol metabolite (M2) and its dimer (M3). Furthermore, a GSH conjugate of M2 (M4) also was formed when the incubation mixture was supplemented with GSH. Experiments with isolated M2 suggested that this metabolite was unstable and underwent spontaneous autooxidation to M3. A qualitatively similar metabolite profile was observed when MRL-A was incubated with recombinant P450 3A4 and cumene hydroperoxide. Because an oxygen atom is transferred to MRL-A under these conditions, these data suggested that S-oxidation alone may result in TZD ring scission and formation of M2 via a sulfenic acid intermediate. Also, because the latter incubation mixture did not contain any reducing agents, the formation of M2 may have occurred due to disproportionation of the sulfenic acid. When NADPH was added to the incubation mixture containing P450 3A4 and cumene hydroperoxide, the formation of M3 increased, suggesting that the sulfenic acid was reduced to M2 by NADPH and subsequently underwent dimerization to yield M3 (vide supra). When NADPH was replaced by GSH, the formation of M4 increased, consistent with reduction of the sulfenic acid by GSH. In summary, these results suggest that the TZD ring in MRL A is activated by an initial P450-mediated S-oxidation step followed by spontaneous scission of the TZD ring to a putative sulfenic acid intermediate; the latter species then undergoes reduction to the free thiol by GSH, NADPH, and/or disproportionation. Finally, the thiol may dimerize to the corresponding disulfide or, in the presence of S-adenosylmethionine, form the stable S-methyl derivative. PMID- 15892583 TI - Formation of tamoxifen-DNA adducts in human endometrial explants exposed to alpha hydroxytamoxifen. AB - An increased risk of developing endometrial cancer has been observed in women receiving tamoxifen (TAM) endocrine therapy and chemoprevention. The genotoxic damage induced by TAM metabolites may be involved in the development of endometrial cancer. To investigate the capability of endometrial tissues to form TAM-DNA adducts, primary cultured human endometrial explants were exposed to alpha-hydroxytamoxifen (alpha-OHTAM) and used for quantitative analysis of TAM DNA adducts, using (32)P-postlabeling/HPLC analysis. A trans isoform of alpha (N(2)-deoxyguanosinyl)tamoxifen (dG-N(2)-TAM) was detected as the major adduct in eight of nine endometrial explants exposed to 100 microM alpha-OHTAM at levels of 7.7 +/- 5.3 (mean +/- SD) adducts/10(7) nucleotides. Approximately 25- and 37 fold lower amounts of the cis form of dG-N(2)-TAM and another trans isoform were also detected. The dG-N(2)-TAM adduct (3.3 adducts/10(7) nucleotides) was detected in one of three endometrial explants exposed to 25 microM alpha-OHTAM. No TAM-DNA adducts were detected in any unexposed tissues. These results indicate that TAM-DNA adducts are capable of forming through O-sulfonation and/or O acetylation of alpha-OHTAM in the endometrium. The endometrial explant culture can be used as a model system to explore the genotoxic mechanism of antiestrogens for humans. PMID- 15892584 TI - Dansyl glutathione as a trapping agent for the quantitative estimation and identification of reactive metabolites. AB - A sensitive and quantitative method was developed for the estimation of reactive metabolite formation in vitro. The method utilizes reduced glutathione (GSH) labeled with a fluorescence tag as a trapping agent and fluorescent detection for quantitation. The derivatization of GSH was accomplished by reaction of oxidized glutathione (GSSG) with dansyl chloride to form dansylated GSSG. Subsequent reduction of the disulfide bond yielded dansylated GSH (dGSH). Test compounds were incubated with human liver microsomes in the presence of dGSH and NADPH, and the resulting mixtures were analyzed by HPLC coupled with a fluorescence detector and a mass spectrometer for the quantitation and mass determination of the resulting dGSH adducts. The comparative chemical reactivity of dGSH vs GSH was investigated by monitoring the reaction of each with 1-chloro-2,4-dinitrobenzene or R-(+)-pulegone after bioactivation. dGSH was found to be equivalent to GSH in chemical reactivity toward both thiol reactive molecules. dGSH did not serve as a cofactor for glutathione S-transferase (GST)-mediated conjugation of 3,4 dichloronitrobenzene in incubations with either human liver S9 fractions or a recombinant GST, GSTM1-1. Reference compounds were tested in this assay, including seven compounds that have been reported to form GSH adducts along with seven drugs that are among the most prescribed in the current U.S. market and have not been reported to form GSH adducts. dGSH adducts were detected and quantitated in incubations with all seven positive reference compounds; however, there were no dGSH adducts observed with any of the widely prescribed drugs. In comparison with existing methods, this method is sensitive, quantitative, cost effective, and easy to implement. PMID- 15892585 TI - Air to blood distribution of volatile organic compounds: a linear free energy analysis. AB - Partition coefficients, K(blood), for volatile organic compounds from air to blood have been collected for 155 compounds (air to human blood) and 127 compounds (air to rat blood). For 86 common compounds, the average error, AE, between the two sets of log K(blood) values is 0.12 log units, somewhat smaller than our estimated interlaboratory average SD value of around 0.16 log units. We conclude that with regard to experimental errors, there is no significant difference between K(blood) values in human blood and in rat blood. There are 196 compounds for which either or both K(blood) (human) and K(blood) (rat) are available. A training set of 98 compounds could be fitted with the Abraham solvation parameters with R(2) = 0.933 and SD = 0.34 log units. The training equation was then used to predict the test set of values with AE = 0.04 log units, SD = 0.33 log units, and an average absolute error, AAE, of 0.25 log units. A second training and test set yielded similar values: AE = 0.01, SD = 0.39, and AAE = 0.29 log units. It is concluded that it is possible to construct an equation capable of predicting further values of log K(blood) to around 0.30 log units. Because the descriptors used in the correlation equations can be predicted from structure, it is now possible to predict log K(blood) for any chemical structure. PMID- 15892586 TI - The appealing story of HIV entry inhibitors : from discovery of biological mechanisms to drug development. AB - Current therapeutic intervention in HIV infection relies upon 20 different drugs. Despite the impressive efficacy shown by these drugs, we are confronted with an unexpected frequency of adverse effects, such as mitochondrial toxicity and lipodystrophy, and resistance, not only to individual drugs but to entire drug classes.Thus, there is now a great need for new antiretroviral drugs with reduced toxicity, increased activity against drug-resistant viruses and a greater capacity to reach tissue sanctuaries of the virus. Two different HIV molecules have been selected as targets of drug inhibition so far: reverse transcriptase and protease. Drugs that target the interactions between the HIV envelope and the cellular receptor complex are a 'new entry' into the scenario of HIV therapy and have recently raised great interest because of their activity against multidrug resistant viruses. There are several compounds that are at different developmental stages in the pipeline to counter HIV entry, among them: (i) the attachment inhibitor dextrin-2-sulfate; (ii) the inhibitors of the glycoprotein (gp) 120/CD4 interaction PRO 542, TNX 355 and BMS 488043; (iii) the co-receptor inhibitors subdivided in those targeting CCR5 (SCH 417690 [SCH D], UK 427857 GW 873140, PRO 140, TAK 220, AMD 887) and those targeting CXCR4 (AMD 070, KRH 2731); and (iv) the fusion inhibitors enfuvirtide (T-20) and tifuvirtide (T-1249). The story of the first of these drugs, enfuvirtide, which has successfully completed phase III clinical trials, has been approved by the US FDA and by the European Medicines Agency, and is now commercially available worldwide, is an example of how the knowledge of basic molecular mechanisms can rapidly translate into the development of clinically effective molecules. PMID- 15892587 TI - Bullous pemphigoid: from bench to bedside. AB - Bullous pemphigoid (BP) is a chronic, autoimmune, blistering disease observed primarily in the elderly population. Several clinical variants have been described, including classic (bullous), localised, nodular, vegetating, erythrodermic, erosive, childhood and drug-induced forms. Autoantibodies target the BP230 and BP180 antigens, located in the hemidesmosomal complex of the skin basement membrane zone. Subsequent complement activation recruits chemical and cellular immune mediators to the skin, ultimately resulting in blister formation. Both autoantibodies and complement may be detected by various immunofluorescent, immune electron microscopy and molecular biology techniques. Recent trials suggest that potent topical corticosteroids should be considered as first-line therapy. Tetracycline with or without nicotinamide may benefit a subset of patients with mild BP. Oral corticosteroids should rarely exceed 0.75 mg/kg/day and corticosteroid-sparing agents may be useful for recalcitrant disease. PMID- 15892588 TI - Antidepressants and sleep: a qualitative review of the literature. AB - Most antidepressants change sleep; in particular, they alter the physiological patterns of sleep stages recorded overnight with EEG and other physiological measures. These effects are greatest and most consistent on rapid eye movement (REM) sleep, and tend to be in the opposite direction to the sleep abnormalities found in major depression, but are usually of greater degree. Reductions in the amount of REM sleep and increases in REM sleep onset latency are seen after taking antidepressants, both in healthy volunteers and in depressed patients. Antidepressants that increase serotonin function by blocking reuptake or by inhibiting metabolism have the greatest effect on REM sleep. The decrease in amount of REM sleep appears to be greatest early in treatment, and gradually diminishes during long-term treatment, except after monoamine oxidase inhibitors when REM sleep is often absent for many months. Sleep initiation and maintenance are also affected by antidepressants, but the effects are much less consistent between drugs. Some antidepressants such as clomipramine and the selective serotonin receptor inhibitors (SSRIs), particularly fluoxetine, are sleep disturbing early in treatment and some others such as amitriptyline and the newer serotonin 5-HT2-receptor antagonists are sleep promoting. However, these effects are fairly short-lived and there are very few significant differences between drugs after a few weeks of treatment. In general, the objectively measured sleep of depressed patients improves during 3-4 weeks of effective antidepressant treatment with most agents, as does their subjective impression of their sleep. Sleep improvement earlier in treatment may be an important clinical goal in some patients, perhaps when insomnia is particularly distressing, or to ensure compliance. In these patients, the choice of a safely used and effective antidepressant which improves sleep in short term is indicated. Patients with other sleep disorders such as restless legs syndrome and REM sleep behaviour disorder should be identified before choosing a treatment, as some antidepressants worsen these conditions. Conversely, there is evidence that some antidepressants may be useful in the treatment of sleep disorders such as night terrors. PMID- 15892590 TI - Tacrolimus: in patients with rheumatoid arthritis. AB - Tacrolimus, a hydrophobic macrolide with immunosuppressant properties, has recently been evaluated as a new treatment for adults with active rheumatoid arthritis. Oral tacrolimus 3mg once daily was significantly more effective than placebo in patients with rheumatoid arthritis (RA) who were refractory or intolerant to disease-modifying antirheumatic drugs (DMARDs), according to results from a 6-month, phase III trial; American College of Rheumatology 20 (ACR20) response rates were 27% and 10%. Tacrolimus 3mg once daily was effective in the same patient group in a 12-month, open-label trial; the ACR20 response rate was 38%. Oral tacrolimus 3 mg once daily was effective in combination with established methotrexate therapy in patients with RA in a 6-month, open-label trial. The ACR20 response rate was 53%. Oral tacrolimus 3 mg once daily was generally well tolerated by patients with active RA refractory or intolerant to previous DMARD treatment or when administered as combination therapy in patients with RA on established methotrexate therapy. PMID- 15892589 TI - Guide to selection of fluoroquinolones in patients with lower respiratory tract infections. AB - Newer fluoroquinolones such as levofloxacin, moxifloxacin, gatifloxacin and gemifloxacin have several attributes that make them excellent choices for the therapy of lower respiratory tract infections. In particular, they have excellent intrinsic activity against Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and the atypical respiratory pathogens. Fluoroquinolones may be used as monotherapy to treat high-risk patients with acute exacerbation of chronic bronchitis, and for patients with community-acquired pneumonia requiring hospitalisation, but not admission to intensive care. Overall, the newer fluoroquinolones often achieve clinical cure rates in > or =90% of these patients. However, rates may be lower in hospital-acquired pneumonia, and this infection should be treated on the basis of anticipated organisms and evaluation of risk factors for specific pathogens such as Pseudomonas aeruginosa. In this setting, an antipseudomonal fluoroquinolone may be used in combination with an antipseudomonalbeta-lactam. Concerns are now being raised about the widespread use, and possibly misuse, of fluoroquinolones and the emergence of resistance among S. pneumoniae, Enterobacteriaceae and P. aeruginosa. A number of pharmacokinetic parameters such as the peak concentration of the antibacterial after a dose (C(max)), and the 24-hour area under the concentration-time curve (AUC24) and their relationship to pharmacodynamic parameters such as the minimum inhibitory and the mutant prevention concentrations (MIC and MPC, respectively) have been proposed to predict the effect of fluoroquinolones on bacterial killing and the emergence of resistance. Higher C(max)/MIC or AUC24/MIC and C(max)/MPC or AUC24/MPC ratios, either as a result of dose administration or the susceptibility of the organism, may lead to a better clinical outcome and decrease the emergence of resistance, respectively. Pharmacokinetic profiles that are optimised to target low-level resistant minor subpopulations of bacteria that often exist in infections may help preserve fluoroquinolones as a class. To this end, optimising the AUC24/MPC or C(max)/MPC ratios is important, particularly against S. pneumoniae, in the setting of lower respiratory tract infections. Agents such as moxifloxacin and gemifloxacin with high ratios against this organism are preferred, and agents such as ciprofloxacin with low ratios should be avoided. For agents such as levofloxacin and gatifloxacin, with intermediate ratios against S. pneumoniae, it may be worthwhile considering alternative dose administration strategies, such as using higher dosages, to eradicate low-level resistant variants. This must, of course, be balanced against the potential of toxicity. Innovative approaches to the use of fluoroquinolones are worth testing in further in vitro experiments as well as in clinical trials. PMID- 15892593 TI - Dexrazoxane : a review of its use for cardioprotection during anthracycline chemotherapy. AB - Dexrazoxane (Cardioxane, Zinecard, a cyclic derivative of edetic acid, is a site specific cardioprotective agent that effectively protects against anthracycline induced cardiac toxicity. Dexrazoxane is approved in the US and some European countries for cardioprotection in women with advanced and/or metastatic breast cancer receiving doxorubicin; in other countries dexrazoxane is approved for use in a wider range of patients with advanced cancer receiving anthracyclines. As shown in clinical trials, intravenous dexrazoxane significantly reduces the incidence of anthracycline-induced congestive heart failure (CHF) and adverse cardiac events in women with advanced breast cancer or adults with soft tissue sarcomas or small-cell lung cancer, regardless of whether the drug is given before the first dose of anthracycline or the administration is delayed until cumulative doxorubicin dose is > or =300 mg/m2. The drug also appears to offer cardioprotection irrespective of pre-existing cardiac risk factors. Importantly, the antitumour efficacy of anthracyclines is unlikely to be altered by dexrazoxane use, although the drug has not been shown to improve progression-free and overall patient survival. At present, the cardioprotective efficacy of dexrazoxane in patients with childhood malignancies is supported by limited data. The drug is generally well tolerated and has a tolerability profile similar to that of placebo in cancer patients undergoing anthracycline-based chemotherapy, with the exception of a higher incidence of severe leukopenia (78% vs 68%; p < 0.01). Dexrazoxane is the only cardioprotective agent with proven efficacy in cancer patients receiving anthracycline chemotherapy and is a valuable option for the prevention of cardiotoxicity in this patient population. PMID- 15892594 TI - Enoxaparin: a review of its use as thromboprophylaxis in acutely ill, nonsurgical patients. AB - Enoxaparin (Clexane), Lovenox) is a low molecular weight heparin (LMWH) that has been widely used in the prevention of venous thromboembolism (VTE) in surgical patients. More recently, with the recognition of the high incidence of VTE in acutely ill medical (nonsurgical) patients, enoxaparin has been evaluated for thromboprophylaxis in this patient population. Subcutaneous enoxaparin 40 mg once daily has shown efficacy in the short-term thromboprophylaxis of VTE in nonsurgical patients with severely restricted mobility due to acute illness in well controlled clinical trials. The drug is at least similar in efficacy to unfractionated heparin (UFH) and its pharmacological profile allows once-daily administration, in contrast to the twice- or three-times-daily administration required with UFH. The tolerability profile of enoxaparin is also similar to that of UFH, except that the incidences of local haematomas and increased liver enzymes are lower with enoxaparin. The optimal duration of prophylaxis in nonsurgical patients is currently being evaluated and the results of extended prophylaxis with enoxaparin evaluated in the EXCLAIM (EXtended CLinical prophylaxis in Acutely Ill Medical patients) trial are awaited with interest. Currently, short-term enoxaparin appears to provide a cost-effective treatment alternative to UFH for VTE prophylaxis in nonsurgical patients. PMID- 15892596 TI - Enhancing patient-based learning: introducing STRAC and the reflective history template. AB - The University of New South Wales School of Rural Health, Australia, like many other rural clinical schools, relies heavily on patient-based learning which is largely serendipitous. The learning is not driven by courses of lectures but by the patients who become available in a variety of settings. Students, who, in their first 3 years have been exposed to a very structured didactic learning experience, start their rural experience in Year IV with a curriculum mainly concerned with general medicine and surgery. They have to learn to adapt to a largely self-directed model. While their rural clinical experience is very much broader than that of their city-based peers, an excellent feature, it engenders feelings of insecurity and a perceived need for students and faculty to have a method of tracking patient encounters so they can ensure the syllabus is covered. Early attempts with a paper-based system and later a Reflective Learning Diary proved unsuccessful. This article presents Syllabus Tracking (STRAC) together with a Reflective Case History Template which encourages reflective learning from patients encountered. STRAC is a web-based solution that includes a reporting function, and is easily modified to suit the needs of the curriculum. It is believed it may also prove useful for future applications. Most rural clinical schools in Australia are young, as are these tools. They are not formally evaluated but are offered to other schools as a suggestion. PMID- 15892597 TI - Design and analysis of phase III trials with ordered outcome scales: the concept of the sliding dichotomy. AB - The conventional approach to the analysis of a Phase III trial in head injury or stroke takes an ordered scale measuring functional outcome and collapses the scale to a binary outcome of favorable versus unfavorable. This discards potentially relevant information which limits statistical power and moreover is not in accord with clinical practice. We propose an alternative approach where a favorable outcome is defined as better than would be expected, taking account of each individual patient's baseline prognosis. This is illustrated through a worked example based on data from a Phase III trial in head injury. The approach is also compared with the proportional odds model, which is another statistical approach that can exploit an ordered outcome scale. The approach raises issues of clinical, statistical, and regulatory importance, and we initiate what we believe needs to become a widespread debate amongst the community involved in clinical research in head injury and stroke. PMID- 15892598 TI - Increased mRNA expression of VEGF within the hematoma and imbalance of angiopoietin-1 and -2 mRNA within the neomembranes of chronic subdural hematoma. AB - The aim of the study was to determine the source of vascular endothelial growth factor (VEGF) in hematoma fluid of patients suffering from chronic subdural hematoma (CSH) and to identify the level of gene expression of the pro-angiogenic factors angiopoietin 1 (ANG-1) and ANG-2 in hematoma membranes. Samples of venous blood, hematoma fluid, and outer membrane were obtained during surgery for CSH. The numbers of mononuclear cells were determined in hematoma fluid and in venous blood samples taken from 11 patients. The concentration of VEGF was measured by ELISA technique in hematoma fluid and in plasma. RT-PCR methodology was used to study the expression of different mRNA species in 11 patients. The mRNA species analyzed include VEGF and its receptors, VEGFR-1 and VEGFR-2, and ANG-1, ANG-2 and their receptor, Tie-2. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as housekeeping gene and was used for semiquantitative analysis. The VEGF concentration was several hundred fold higher in the hematoma fluid than in corresponding plasma samples. A significant correlation was found between the number of neutrophils and the VEGF content in the hematoma fluid. The expression levels of VEGF, mainly VEGF165 and VEGF121 mRNA were highest in cells obtained from the hematoma fluid. In membrane samples, mRNA encoding for VEGF and its receptors was only inconsistently detected while the mRNA species encoding for ANG-1, ANG-2, and Tie-2 were found throughout all samples. The mean ratio of ANG 1/ANG-2 mRNA expression was 0.48 as opposed to 1.9 in a normal human brain tissue sample. The results suggest that the hematoma cells are the primary source of VEGF. A marked increase in the expression of ANG-2 mRNA over ANG-1 mRNA demonstrates a pro-angiogenic pattern in the hematoma membranes. Persistent activation of the ANG/Tie-2 system in addition to high levels of VEGF may keep the vasculature in a destabilized condition and may account for the continuous formation of new and immature blood vessels resulting in massive plasma extravasation and repeated bleeding episodes. Thus, the present study provides new evidence in favor of pro-angiogenic mechanisms playing an important role in the pathophysiology of CSH. PMID- 15892599 TI - Functional consequences of lumbar spinal cord contusion injuries in the adult rat. AB - Our understanding of the substrates of locomotion, and hence our understanding of the causes of deficits following spinal cord injury, is still incomplete. While severe locomotor deficits can be induced by either contusion or laceration injuries or demyelination of thoracic spinal cord ventral and ventrolateral white matter, loss of mid-thoracic gray matter (intraspinal kainic acid injection) has no impact on locomotion. In contrast, loss of gray matter from the rostral lumbar segments induces severe locomotor deficits. This study examines the histological and locomotor outcomes following contusion injuries involving the rostral segments of the lumbar enlargement in the adult rat. Adult Sprague-Dawley rats received contusion injuries centered on the T13/L1, L2, or L3/4 spinal cord segments. Moderately severe injuries centered on the T13/L1 and L2 spinal cord segments induced more severe locomotor deficits than those centered on the L3/4 segments, despite a significantly smaller total gray matter volume loss (1.7 vs. 2.7 mm3). Moderately-severe injuries at T13/L1, L2, and L3/4 showed 21%, 31%, and 39% white matter sparing, respectively, with 6-week BBB scores of 10, 10, and 15.7, respectively. These data suggest that moderately-severe contusion injuries centered on the rostral segments of the lumbar enlargement induce more severe locomotor deficits than would be predicted by the histological outcome (spared white matter), suggesting that gray matter loss may play a role in functional deficits following some lumbar contusion injuries. PMID- 15892600 TI - Motoneuron loss associated with chronic locomotion impairments after spinal cord contusion in the rat. AB - Information on the nature of deficits and adaptive mechanisms occurring after spinal cord injury is essential to the design of strategies for promoting functional recovery. Motor impairments and compensations were quantified by three dimensional kinematic analysis in freely walking rats, 6 months after mild cervical (C7) or moderate lumbar (L2) spinal cord contusion. After C7 contusion, the animals showed reduced elbow extension and wrist movement, whereas reduced knee extension was the main impairment after L2 contusion. In both cases, the duration of the walking cycle increased and forward velocity was reduced due to a longer stance phase. Histology revealed reproducible lesions extending approximately to one spinal cord segment. In the transverse plane, the lesion involved the central gray matter and adjacent axons, including the dorsal corticospinal tract, but partially spared the ventrolateral tracts. Retrograde motoneuron tracing by nerve exposure to HRP or intramuscular injection of aminostilbamidine demonstrated that C7 contusion caused the loss of approximately 40% of triceps brachii motoneurons, whereas approximately 30% of quadriceps femoris motoneurons were lost after L2 contusion. These results demonstrate permanent deficits after incomplete lesions at the spinal cord enlargements and suggest that motoneuron loss contributes to their production. PMID- 15892601 TI - Rapid functional recovery after spinal cord injury in young rats. AB - Responses to traumatic injury in the immature spinal cord may be different from those in adults. We modified an adult model of weight-drop injury to characterize the histopathology and functional recovery after spinal cord injury (SCI) in rat pups at postnatal day 14-15. A 10-g weight was dropped from 2.5 or 5.0 cm at T8 T9. Hindlimb function was evaluated at 24 h and 1, 2, 3, and 4 weeks after injury using the Combined Behavioral Score that estimates overall hind limb sensorimotor function, and the BBB scale for open field locomotion. Histopathology was examined at 15 min, 24 h, and 4 weeks after SCI. The initial hemorrhagic lesion was similar to that seen in adults, but the time course of secondary loss of ventral horn motor neurons was extended. By 4 weeks, only a partial rim of white matter surrounding a central cavity was seen. The 5.0 cm injury group exhibited significantly less recovery of function at 4 weeks than the 2.5 cm group. In the latter, the degree of hindlimb deficit at 4 weeks was similar to that previously described for adults with 10 g x 2.5 cm SCI. However, pups in both injury groups exhibited a significantly faster rate of recovery than adults. Recovery was maximal by 1 week after SCI in pups as compared to 3-4 weeks in adults. The more rapid functional recovery observed in the pups suggests that this new model may be useful for studying mechanisms of functional plasticity after SCI. PMID- 15892602 TI - Effects of glial transplantation on functional recovery following acute spinal cord injury. AB - Numerous efforts have been made to maximize the efficacy of treatment for spinal cord injury (SCI). Recently, oligodendrocyte-type 2 astrocyte (O-2A) progenitor cells have been reported to remyelinate focal areas of demyelinated spinal cord in adult rats. We conducted a study to investigate the therapeutic potential of transplantation of O-2A cells in a rat model of acute SCI. SCI was induced with an NYU Impactor at T9 of rats. O-2A cells labeled with bromodeoxyuridine (BrdU) were transplanted into sites of SCI at 1 week after the induction of SCI. At 6 weeks after cell transplantation, a behavioral test showed significant functional improvement in animals that had received O-2A-cell transplants as compared to animals given cell-culture medium alone. An electrophysiological study revealed that the transplants did not improve the amplitude or latency of somatosensory evoked potentials, but a recording of motor evoked potentials showed that the latency of these potentials in the O-2A-cell-transplant group was significantly shorter than that in the group treated with cell-culture medium. Following transplantation of BrdU-labeled O-2A cells, cells positive for BrdU were detected at and near sites of SCI. Cells labeled for both BrdU and 2',3' -cyclic nucleotide-3-phosphodiesterase were also detected, showing that the transplanted O-2A cells differentiated into oligodendrocytes. By contrast, cells labeled for BrdU and glial fibrillary acidic protein, or for neuronal nuclei antigen, were not detected. Furthermore, a tract-tracing study showed that numbers of retrogradely labeled neurons increased in areas of the brain stem after O-2A-cell transplantation. The study data showed that after being transplanted into an animal with SCI, O-2A cells migrated to the area adjacent to the site of injury and differentiated into oligodendrocytes. The behavioral test and the electrophysiological and morphological studies showed that transplantation of O 2A cells may play an important role in functional recovery and the regeneration of axons after SCI. PMID- 15892603 TI - Serotonin-related enhancement of recovery of hind limb motor functions in spinal rats after grafting of embryonic raphe nuclei. AB - Recently, we demonstrated improvements in hind limb locomotor-like movements following grafting of embryonic raphe nuclei cells into the spinal cord below the level of total transection in adult rats. The purpose of the present study was to clarify whether this improvement was due to newly established serotonergic innervation between the graft and the host. Two months after intraspinal grafting of the embryonic raphe nuclei, the spinalized rats, when put on a treadmill, could be induced to walk with regular alternating hind limb movements with the plantar contact with the ground during the stance phase, and ankle dorsiflexion during the swing phase of each step cycle. In the same situation the spinal rats, that did not receive the graft, were not able to initiate the dorsiflexion of the ankle joint during the swing phase and very often the dorsal surface of the foot was dragged along the ground. Intraperitoneal application of directly acting 5 HT2 antagonist Cyproheptadine (1 mg/kg) impaired reversibly the hind limb locomotor-like movements in grafted rats. This impairment lasted for 2-3 h. The same procedure in control rats did not markedly alter the hind limb locomotor like movements. The effect of Cyproheptadine in grafted rats was reversed by i.p. injections of the 5-HT2 agonist Quipazine (0.5 mg/kg). These results show that the graft-induced restitution of hind limb locomotor abilities in adult spinal rats is brought about by the new serotonergic innervation of the host spinal cord circuitry from the grafted neurons and is mediated by 5-HT2 receptors. PMID- 15892604 TI - Origin of regenerated axons in nerve bypass grafts. AB - It has been shown that end-to-side coaptation and nerve bypass grafting, which are essentially two sequential end-to-side coaptations, induce axonal outgrowth in peripheral nerve injuries. However, it is unknown whether the axons regenerated after end-to-side coaptation originate by collateral sprouting at the suture site or by elongation from the spinal neuronal pool. Also unknown is the extent of functional recovery that can be expected after bypass grafting for the reconstruction of injured peripheral nerves. We conducted a study to evaluate the origin of regenerated axons after end-to-side coaptation and the utility of nerve bypass grafting for peripheral nerve injury. For this purpose, we performed electrophysiological studies using bypass grafting (end-to-side coaptation) and conventional cable grafting (end-to-end coaptation) to treat complete paralysis of the peroneal nerve in rabbit models, and compared the recovery time and extent of functional recovery achieved with the two techniques. We assessed, by electromyography, the time to appearance of reinnervation potentials from the tibialis anterior muscle on the affected side. These times were not significantly different in the two study groups of animals (p = 0.5390). After a 12-week recovery period, electrophysiological findings and histological assessment showed similar recovery in both groups of animals. It is known that collateral sprouting of axons from the nodes of Ranvier proximal to the transected nerve stump occurs in cable grafting, and that axon elongation from the spinal cord requires more time. Our findings in the present study strongly suggest that collateral sprouting across end-to-side sutures is the chief means of axonal outgrowth in nerve bypass grafts, and that functional recovery can be expected in bypass grafting to nearly the same extent as in cable grafting. PMID- 15892605 TI - Comparative analysis of transcription profiles of Helicobacter pylori clinical isolates. AB - The transcription profiles of four Helicobacter pylori clinical isolates (two cag negative and two cag-positive) were compared in stationary growth phase using a cDNA-macroarray. The correlation coefficient value between total transcription profiles of clinical isolates H. pylori varied from 0.70 to 0.83. For 44 groups of genes (total number 66) belonging to various functional classes of H. pylori, the correlation coefficient value between these isolates exceeded 0.7, and for 14 groups the value exceeded 0.9. These groups included genes encoding components involved in cell division, adaptations to atypical conditions, electron transport, salvage of nucleosides and nucleotides, glycolysis/gluconeogenesis, folding and stabilization of proteins, translation factors, anaerobic metabolism, and amino acids and amine metabolism. Expression of 52 genes significantly differed between H. pylori clinical isolates. Some of these genes determine microorganism virulence. They include: cytotoxin-associated gene (cagA), genes encoding neutrophil-activating protein (napA), major flagellar protein (flaA), and vacuolizing cytotoxin (vacA), some genes encoding outer membrane proteins (omp), urease alpha and beta subunits (ureA and ureB), and some regulatory proteins, and genes encoding stress-related proteins, such as the chaperone and heat shock protein genes (groEL and dnaK). PMID- 15892606 TI - Molecular mechanisms of hormonal activity. II. Kinase systems. Systems with intracellular receptors. Transactivation of STS. AB - Hormone receptors and other components, functional mechanisms, and biological role of analyzed signal transduction systems (STS) are described. The recently revealed module principle of the structure and STS transactivation providing diversity and plasticity of regulation are highlighted. STS activities are significantly changed in many diseases. Novel promising pharmaceuticals targeted to certain components of STS increase in number from year to year. The data published by the beginning of January 2004 are summarized in this review. PMID- 15892607 TI - Molecular mechanisms of epigenetics. AB - The main epigenetic mechanisms in regulation of gene expression are discussed. The definition of epigenetics and its specific mechanisms including DNA methylation and gene imprinting, modifications of nucleosomal histones associated with silencing or activation of gene transcription, RNA interference, chromosomal silencing, and the role of mobile elements are discussed. PMID- 15892608 TI - Possible reasons for difference in sensitivity to oxygen of two Escherichia coli strains. AB - In preliminary experiments it was found that Escherichia coli strains AB1157 and KS400 are different in their abilities to grow under various oxygen levels in cultivation medium: the first strain does not grow under high oxygen conditions, unlike the second one. To investigate whether the damage to cellular components due to production of reactive oxygen species (ROS) was responsible for this difference, the intensity of free radical oxidation of proteins and lipids as well as the activities of selected antioxidant and associated enzymes (superoxide dismutase, catalase, peroxidase, glutathione reductase, glucose-6-phosphate dehydrogenase) were compared in the two strains. The level of thiobarbituric acid reactive substances was 1.8-2.5-fold higher in AB1157 than in KS400, but the concentration of carbonyl proteins was lower in the AB1157 strain. In both strains growth under higher oxygen levels resulted in higher superoxide dismutase and peroxidase activities in both exponential and stationary phases. Overall, the activities of antioxidant enzymes were always higher in the KS400 strain than in AB1157. The results for both lipid and protein oxidative damage and antioxidant enzyme activities suggest that the differences in oxygen tolerance between these two strains may be due to their different abilities to cope with ROS. PMID- 15892609 TI - Monomeric and multimeric blockers of selectins: comparison of in vitro and in vivo activity. AB - The potency of the oligosaccharides SiaLe(x), SiaLe(a), HSO(3)Le(x), and HSO(3)Le(a), their conjugates with polyacrylamide (PAA, 40 kD), and other monomeric and polymeric selectin inhibitors has been compared with that of the polysaccharide fucoidan. The following assay systems were used: 1) a 96-well assay based either on the use of recombinant E-, P-, and L-selectins or an analogous assay with natural P-selectin isolated from human platelets; 2) a platelet-based P-selectin cell assay; and 3) a rat model of peritoneal inflammation. IC(50) values for the neoglycoconjugate SiaLe(a)-PAA were 6, 40, and 85 microM for recombinant E-, P-, and L-selectins, respectively; all monomeric inhibitors were about two orders of magnitude weaker. PAA-conjugates, containing as a ligand tyrosine-O-sulfate (sTyr) in addition to one of the sialylated oligosaccharides, were the most potent synthetic blockers in vitro. Compared with fucoidan, the most potent known P- and L-selectin blocker, the bi ligand glycoconjugate HSO(3)Le(a)-PAA-sTyr displayed similar inhibitory activity in vitro towards L-selectin and about ten times lower activity towards P selectin. All of the tested synthetic polymers displayed a similar ability to inhibit neutrophil extravasation in the peritonitis model (in vivo) at 10 mg/kg. The data provide evidence that monomeric SiaLe(x) is considerably more effective as a selectin blocker in vivo than in vitro, whereas the opposite is true for fucoidan and the bi-ligand neoglycoconjugate HSO(3)Le(a)-PAA-sTyr. PMID- 15892610 TI - Oscillatory activity of P-type membrane adenosine triphosphatases: a kinetic model. AB - A kinetic model for membrane P-type adenosine triphosphatases is considered, the main application being to the erythrocyte Ca2+-ATPase. It is shown that a simple modification of the known catalytic mechanism of the ATPase by addition of a self inhibition step and the steady calcium influx leads to damped oscillations in the system discussed. In this way, the model can explain the kinetic experimental results obtained for the purified enzyme in solution as well as for the enzyme incorporated into liposome membranes. The estimated kinetic parameters are close to the experimental ones. Alternative changes in time, demonstrated by the kinetic model for the conformational enzyme states, E(1 )and E(2), confirm the model of two alternatively functioning gates in the ion pumping Ca2+-ATPase. PMID- 15892611 TI - Influence of Ca2+ oscillatory influx on membrane Ca2+-ATPase activity: a kinetic model. AB - A kinetic model for the membrane Ca2+-ATPase is considered. The catalytic cycle in the model is extended by enzyme auto-inhibition and by oscillatory calcium influx. It is shown that the conductive enzyme activity can be registered as damped or sustained Ca2+ pulses similar to observed experimentally. It is shown that frequency variations in Ca2+ oscillatory influx induce changes of pulsating enzyme activity. Encoding is observed for the signal frequency into a number of fixed levels of sustained pulses in the enzyme activity. At certain calcium signal frequencies, the calculated Ca2+-ATPase conductivity demonstrates chaotic multi-level pulses, similar to those observed experimentally. PMID- 15892612 TI - Study of applicability of the bifunctional system "ethidium bromide + Hoechst 33258" for DNA analysis. AB - Changes in absorbance and fluorescence excitation and emission spectra in the ultraviolet and visible regions of the system containing ethidium bromide (EtBr) and Hoechst-33258 (Ht) were investigated depending on various DNA quantities and the composition of the medium. It is noted that spectral properties of this system are determined by interactions of EtBr and Ht both with nucleic acid and with one another (for example, joint sorption of EtBr and Ht on DNA may involve fluorescent resonance energy transfer between the dye molecules). Thus, different modes of EtBr and Ht specificity to substrate and assay conditions suggest that combined use of these dyes provides some additional effects that may be interesting in terms of structure-functional study of nucleic acid. Some of these effects are considered in this paper. PMID- 15892613 TI - Human immunoglobulin light chains lambda form amyloid fibrils and granular aggregates in solution. AB - Myeloma nephropathy is a disorder characterized by deposition of monoclonal immunoglobulin light chains in the kidneys. The chains deposited form either amyloid fibrils or granular (amorphous) aggregates. Distinct molecular mechanisms leading to the formation of different aggregate types in kidney of patients with multiple myeloma are poorly understood. Here we describe the self-association kinetics of human monoclonal immunoglobulin light chains lambda (GRY) isolated from urine of a patient with multiple myeloma. Under physiological conditions, the isolated light chain exists predominantly in a form of covalent dimer with apparent molecular mass of 50.1 kD. Spectral probe binding, analytical gel filtration, Western blot analysis, and electron microscopy indicate that GRY dimer aggregation occurs via two different pathways producing either amyloid fibrils or amorphous aggregates depending on microenvironment. Incubation of GRY (25 microM) for 4-14 days at 37 degrees C in phosphate buffered saline (PBS), pH 7.0, or in PBS containing urea (0.8 M), pH 6.5, leads to amyloid fibril formation. Under electron microscopy, the fibrils show unbranched thread-like structures, approximately 60-80 x 1000 A in size, which can bind thioflavin T and Congo Red. GRY maintained in acetate buffer, pH 3.5, forms granular aggregates. The structure of GRY oligomers formed during the early stage of amyloid fibril formation (1-4 days) has been examined by means of protein cross-linking with homobifunctional reagents. These oligomers are predominantly trimers and tetramers. PMID- 15892614 TI - Oxidase reaction of the hybrid Mn-peroxidase of the fungus Panus tigrinus 8/18. AB - The hybrid Mn-peroxidase of the fungus Panus tigrinus 8/18 oxidized NADH in the absence of hydrogen peroxide, this being accompanied by the consumption of oxygen. The reaction of NADH oxidation started after a period of induction and completely depended on the presence of Mn(II). The reaction was inhibited in the presence of catalase and superoxide dismutase. Oxidation of NADH by the enzyme or by manganese(III)acetate was accompanied by the production of hydrogen peroxide and superoxide radicals. In the presence of NADH, the enzyme was transformed into a catalytically inactive oxidized form (compound III), and the latter was inactivated with bleaching of the heme. The substrate of the hybrid Mn-peroxidase (Mn(II)) reduced compound III to yield the native form of the enzyme and prevented its inactivation. It is assumed that the hybrid Mn-peroxidase used the formed hydrogen peroxide in the usual peroxidase reaction to produce Mn(III), which was involved in the formation of hydrogen peroxide and thus accelerated the peroxidase reaction. The reaction of NADH oxidation is a peroxidase reaction and the consumption of oxygen is due to its interaction with the products of NADH oxidation. The role of Mn(II) in the oxidation of NADH consisted in the production of hydrogen peroxide and the protection of the enzyme from inactivation. PMID- 15892615 TI - Proposed mechanism of nitrite-induced methemoglobinemia. AB - A scheme of development of nitrite-induced oxyhemoglobin oxidation in erythrocytes based on the analysis of experimental data is proposed. It was found that, contrary to widespread opinion, direct oxidative-reductive interaction between hemoglobin and nitrite is absent or negligible under physiological conditions. The driving stage of this process is methemoglobin-catalyzed peroxidase oxidation of nitrite. The product of the oxidation (presumably NO2*) directly oxidizes oxyhemoglobin to methemoglobin-peroxide complex without hydrogen peroxide release into the environment. The oxidant itself is reduced to nitrite or oxidized to nitrate as a result of interaction with another NO2* molecule. Thus, the stoichiometry of the process depends on the ratio of rates of these two reactions. Substances that are able to compete with nitrite for peroxidase and therefore to prevent the nitrite oxidation effectively protect hemoglobin from oxidation. Catalase is not able to destroy methemoglobin-peroxide complexes, but it can prevent their production in the course of interaction of methemoglobin and free peroxide by destroying the latter. PMID- 15892616 TI - EphA2 receptor tyrosine kinase as a promising target for cancer therapeutics. AB - Eph receptors are a unique family of receptor tyrosine kinases (RTK) that play critical roles in embryonic patterning, neuronal targeting, and vascular development during normal embryogenesis. Eph RTKs and their ligands, the ephrins, are also frequently overexpressed in a variety of cancers and tumor cell lines. In particular, one family member, EphA2, is overexpressed in breast, prostate, lung, and colon cancers. Unlike traditional oncogenes that often function only in tumor cells, recent data show that Eph receptors mediate cell-cell interactions both in tumor cells and in the tumor microenvironment, namely the tumor stroma and tumor vasculature. Thus, EphA2 receptors are attractive targets for drug design, as targeting these molecules could simultaneously inhibit several aspects of tumor progression. This review focuses on the multiple roles of EphA2 in cancer progression, the mechanisms by which EphA2 inhibition may halt this progression, and the pre-clinical results of EphA2 inhibition in various cancer model systems. PMID- 15892617 TI - Modulation of pRB/E2F functions in the regulation of cell cycle and in cancer. AB - Cell proliferation is regulated by the cell cycle, and in order to divide the cell must enter a mitotic state. Prior to mitosis the cell is required to pass through a number of checkpoints, including the critical G1/S restriction point governed by the successive phosphorylation of the retinoblastoma protein, pRb. The various proteins and regulatory factors governing pRb phosphorylation have been a major focus of study in recent years, given the central importance of G1/S transition deregulation in cancer development. This review summarises the molecular biology around the G1/S transition, focussing on the critical roles of the transcription factor family E2F and the cyclin-dependent kinase (CDK) and cyclin families involved in E2F release from pRb. Interestingly, E2F release from pRb is associated with cell proliferation; however, above a certain threshold E2F has the potential to trigger apoptosis. The review focuses on the following topics: (i) how E2F and other substrates bind to pRb at the molecular level; (ii) mechanisms by which pRb function is modulated within the cell; (iii) mechanisms that inhibit or enhance cell proliferation via the pRb/E2F pathway; (iv) how E2F can potentiate apoptotic pathways; and (v) what controls whether E2F mediates cell proliferation or apoptosis. The case for the development of agents that perturb pRb:E2F interactions will be made, as a strategy to further inform the molecular biology around this important target and as a therapeutic strategy against cancer. PMID- 15892618 TI - Molecularly targeted therapy for gastrointestinal cancer. AB - Receptor and non-receptor tyrosine kinases (TKs) have emerged as clinically useful drug target molecules for treating gastrointestinal cancer. Imatinib mesilate (STI-571, Gleevec(TM)), an inhibitior of bcr-abl TK, which was primarily designed to treat chronic myeloid leukemia is also an inhibitor of c-kit receptor TK, and is currently the drug of choice for the therapy of metastatic gastrointestinal stromal tumors (GISTs), which frequently express constitutively activated forms of the c-kit-receptor. The epidermal growth factor receptor (EGFR), which is involved in cell proliferation, metastasis and angiogenesis, is another important target. The two main classes of EGFR inhibitors are the TK inhibitors and monoclonal antibodies. Gefitinib (ZD1839, Iressa(TM)) has been on trial for esophageal and colorectal cancer (CRC) and erlotinib (OSI-774, Tarceva(TM)) on trial for esophageal, colorectal, hepatocellular, and biliary carcinoma. In addition, erlotinib has been evaluated in a Phase III study for the treatment of pancreatic cancer. Cetuximab (IMC-C225, Erbitux(TM)), a monoclonal EGFR antibody, has been FDA approved for the therapy of irinotecan resistant colorectal cancer and has been tested for pancreatic cancer. Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) are critical regulators of tumor angiogenesis. Bevacizumab (Avastin(TM)), a monoclonal antibody against VEGF, was efficient in two randomized clinical trials investigating the treatment of metastatic colorectal cancer. It is also currently investigated for the therapy of pancreatic cancer in combination with gemcitabine. Other promising new drugs currently under preclinical and clinical evaluation, are VEGFR2 inhibitor PTK787/ZK 222584, thalidomide, farnesyl transferase inhibitor R115777 (tipifarnib, Zarnestra(TM)), matrix metalloproteinase inhibitors, proteasome inhibitor bortezomib (Velcade(TM)), mammalian target of rapamycin (mTOR) inhibitors, cyclooxygenase-2 (COX-2) inhibitors, platelet derived growth factor receptor (PDGF-R) inhibitors, protein kinase C (PKC) inhibitors, mitogen activated protein kinase kinase (MEK) 1/2 inhibitors, Rous sarcoma virus transforming oncogene (SRC) kinase inhibitors, histondeacetylase (HDAC) inhibitors, small hypoxia-inducible factor (HIF) inhibitors, aurora kinase inhibitors, hedgehog inhibitors, and TGF-beta signalling inhibitors. PMID- 15892619 TI - Copper lowering therapy with tetrathiomolybdate as an antiangiogenic strategy in cancer. AB - Tetrathiomolybdate (TM) is a novel anticopper agent under development for use in Wilson's disease. It acts by forming a stable tripartite complex with serum albumin and copper, rendering the complexed copper unavailable for cellular uptake. TM is a very potent anticopper agent and has an excellent safety profile. It has been shown that normal copper levels are required for optimal angiogenesis. Based on this background, we decided to evaluate TM as an anticancer agent. TM treatment of Her/2neu mice, genetically programmed to develop breast cancer, completely prevented the development of visible mammary cancers, although avascular microscopic clusters of cancer cells were present in the breasts of TM treated animals. Controls developed grossly visible tumors. TM was able to strongly inhibit tumor growth in six other rodent models. In a phase 1/2 clinical trial of advanced and metastatic cancers, freedom from progression averaged 11 months, and some individual results were quite dramatic. Eight phase 2 studies of specific cancers have been launched. TM's hypothesized mechanism of action is inhibition of angiogenic cytokines. Unlike other current approaches to antiangiogenic therapy which target single agents, we hypothesize that TM inhibits multiple angiogenic cytokines. Part of this effect appears to stem from inhibition of nuclear factor kappa B (NF(K)B), which in turn controls transcription of many angiogenic and other cytokines. However, there are probably multiple mechanisms, in that some angiogenic cytokines appear to have separate mechanisms of copper dependence. The inhibition of multiple angiogenic cytokines gives TM the potential to be a more global inhibitor of angiogenesis. PMID- 15892620 TI - Matrix metalloproteinases as therapeutic targets in cancer. AB - Degradation of extracellular matrix is crucial for malignant tumor growth, invasion, metastasis and angiogenesis. Matrix metalloproteinases (MMPs) are a family of zinc-dependent neutral endopeptidases collectively capable of degrading essentially all matrix components. Elevated levels of distinct MMPs can be detected in tumor tissue or serum of patients with advanced cancer, and their role as prognostic indicators in cancer has been widely examined. In addition, therapeutic intervention in tumor growth and invasion based on inhibition of MMP activity is under intensive investigation and several MMP inhibitors (MMPIs) are in clinical cancer trials. Even though results of the first clinical trials in advanced cancer have been mostly disappointing, there are also positive results. Recent observations show, that certain MMPs limit tumor growth. Therefore, identification of proper MMPs for therapeutic intervention with array-based molecular classifications of tumors and targeting these with more specific MMPIs in combination with conventional chemotherapy is expected to provide a feasible approach for cancer therapy. MMPIs represent a totally different therapeutic modality from proven anti-cancer drugs and thus traditional approaches to evaluate drug efficiency cannot be used without modification. In this review, we discuss the current view on the feasibility of MMPs as targets for therapeutic intervention in cancer. PMID- 15892621 TI - New target antigens for cancer immunoprevention. AB - Prevention of cancer through the activation of the immune system has been explored in recent years in preclinical systems thanks to the availability of several new transgenic mouse models that closely mimic the natural history of human tumors. The most thoroughly investigated model of cancer immunoprevention is the mammary carcinoma of HER-2/neu transgenic mouse. In this system it has clearly been shown that the activation of immune defences in healthy individuals can effectively prevent the subsequent onset of highly aggressive mammary carcinomas. A complete prevention was obtained using a combination of three signals (the so called "triplex" vaccine) that included the specific antigen (p185, the product of HER-2/neu) and nonspecific signals like allogeneic histocompatibility antigens and interleukin 12. The analysis of protective immune responses in models of cancer immunoprevention revealed some unexpected features, in particular the central role of antibodies in immunoprevention, at variance with conventional immuno-therapy which is firmly based on cytotoxic T cells. In the HER-2/neu system anti-p185 antibodies, in addition to immunological functions leading to tumor cell lysis, inhibit p185 dimerization and induce its internalization, resulting in the inhibition of mitogenic signaling. Most current tumor antigens appear to be unsuitable targets for cancer immunoprevention. An ideal antigen should have a crucial pathogenetic role in tumor growth to avoid the selection of antigen loss variants. Downregulation of major histocompatibility complex (MHC) expression during tumor progression frequently limits antigen recognition by MHC-restricted T cells. Thus an ideal antigen for cancer immunoprevention should be recognized both by T cells and by antibodies. Antibody binding to cell surface oncogenic determinants, in addition to complement- and cell-mediated tumor cell lysis, can block mitogenic signaling and induce internalization, resulting in tumor growth arrest. A search for new tumor antigens should be conducted among molecules that are directly involved in neoplastic transformation and are recognizable by the immune response also in MHC loss variants. Novel tumor antigens fulfilling both conditions will be crucial for the development of cancer immunoprevention and will provide new targets also for cancer immunotherapy. PMID- 15892623 TI - Defensins and other antimicrobial peptides: a historical perspective and an update. AB - Antimicrobial peptides are effectors of innate immunity in phagocytes, body fluids and epithelia. In mammals, defensins, peptides with a characteristic six cysteine framework, are particularly abundant and widely distributed in various animal species and tissues. The first part of this review provides a historical overview of the ideas that led to the current state-of-the-art in antimicrobial peptides, and the second part is an update on mammalian defensins and their role in host defense to infections. PMID- 15892624 TI - Improving on nature's defenses: optimization & high throughput screening of antimicrobial peptides. AB - Antimicrobial peptides (AMPs) are ubiquitous in nature where they play important roles in host defense and microbial control. Despite their natural origin, antimicrobial spectrum and potency, the lead peptide candidates that so far have entered pharmaceutical development have all been further optimized by rational or semi-rational approaches. In recent years, several high throughput screening (HTS) systems have been developed to specifically address optimization of AMPs. These include a range of computational in silico systems and cell-based in vivo systems. The in silico-based screening systems comprise several computational methods such as Quantitative Structure/Activity Relationships (QSAR) as well as simulation methods mimicking peptide/membrane interactions. The in vivo-based systems can be divided in cis-acting and trans-acting screening systems. The cis acting pre-screens, where the AMP exerts its antimicrobial effect on the producing cell, allow screening of millions or even billions of lead candidates for their basic antimicrobial or membrane-perturbating activity. The trans-acting screens, where the AMP is secreted or actively liberated from the producing cell and interacts with cells different from the producing cell, allow for screening under more complex and application-relevant conditions. This review describes the application of HTS systems employed for AMPs and lists advantages as well as limitations of these systems. PMID- 15892625 TI - Peptoids as source of compounds eliciting antibacterial activity. AB - N-Alkylglycine oligomers (peptoids) constitute a family of non-natural peptidomimetics attractive for the early drug discovery process because of their physicochemical features, easy of adaptation to combinatorial chemistry approaches and their proteolytic stability. Consequently, peptoid libraries have found application for discovering hits against a wide diversity of pharmaceutical targets, among which different examples of antibacterials are found. In the present work, research efforts addressed towards the identification of peptoids as antibacterial agents are discussed. PMID- 15892626 TI - Molecular mechanisms of membrane perturbation by antimicrobial peptides and the use of biophysical studies in the design of novel peptide antibiotics. AB - Antibiotic resistant bacterial strains represent a global health problem with a strong social and economic impact. Thus, there is an urgent need for the development of antibiotics with novel mechanisms of action. There is currently an extensive effort to understand the mode of action of antimicrobial peptides which are considered as one alternative to classical antibiotics. The main advantage of this class of substances, when considering bacterial resistance, is that they rapidly, within minutes, kill bacteria. Antimicrobial peptides can be found in every organism and display a wide spectrum of activity. Hence, the goal is to engineer peptides with an improved therapeutic index, i.e. high efficacy and target specificity. For the rational design of such novel antibiotics it is essential to elucidate the molecular mechanism of action. Biophysical studies have been performed using to a large extent membrane model systems demonstrating that there are distinctive different mechanisms of bacterial killing by antimicrobial peptides. One can distinguish between peptides that permeabilize and/or disrupt the bacterial cell membrane and peptides that translocate through the cell membrane and interact with a cytosolic target. Lantibiotics exhibit specific mechanisms, e.g. binding to lipid II, a precursor of the peptidoglycan layer, either resulting in membrane rupture by pore formation or preventing cell wall biosynthesis. The classical models of membrane perturbation, pore formation and carpet mechanism, are discussed and related to other mechanisms that may lead to membrane dysfunction such as formation of lipid-peptide domains or membrane disruption by formation of non-lamellar phases. Emphasis is on the role of membrane lipid composition in these processes and in the translocation of antimicrobial peptides. PMID- 15892627 TI - Design of host defence peptides for antimicrobial and immunity enhancing activities. AB - Host defense peptides are a vital component of the innate immune systems of humans, other mammals, amphibians, and arthropods. The related cationic antimicrobial peptides are also produced by many species of bacteria and function as part of the antimicrobial arsenal to help the producing organism reduce competition for resources from sensitive species. The antimicrobial activities of many of these peptides have been extensively characterized and the structural requirements for these activities are also becoming increasingly clear. In addition to their known antimicrobial role, many host defense peptides are also involved in a plethora of immune functions in the host. In this review, we examine the role of structure in determining antimicrobial activity of certain prototypical cationic peptides and ways that bacteria have evolved to usurp these activities. We also review recent literature on what structural components are related to these immunomodulatory effects. It must be stressed however that these studies, and the area of peptide research, are still in their infancy. PMID- 15892628 TI - Cathelicidins--nature's attempt at combinatorial chemistry. AB - Cathelicidins are a family of diverse antimicrobial peptides found in granules of mammalian neutrophils. Cathelicidins are active against a broad range of microbes in different environments. Aside from their antimicrobial activity, cathelicidins possess other biological properties including cytotoxic activity towards mammalian cells. Several studies have shown that the amino acid sequence of cathelicidins can be modified to temper undesired properties, such as hemolytic and cytotoxic activity, and at the same time maintain antimicrobial activity. These properties make cathelicidins ideal templates in combinatorial chemistry for designing de novo antimicrobial peptides for therapeutic use. However, one of the major challenges will be to screen these peptides in experimentally relevant models that reflect the environments in which the peptides should be therapeutically active. PMID- 15892629 TI - Therapeutic opportunities in Alzheimer disease: one for all or all for one? AB - In recent years, Alzheimer disease (AD) has received great attention as an incurable and fatal disease that threatens the lives of aging individuals. Debates regarding areas of research and treatment designs have made headlines as scientists in the field question ongoing work. Despite these academic quarrels, significant insights concerning the cellular and molecular basis of AD have illuminated the potential causes and consequences of AD pathogenesis in the human brain. Additionally, assigning relationships among scientific evidence is difficult due to the nature of the disease. It is crucial to note that all findings do not constitute causality as AD has many stages of progression, and therefore a particular finding may reflect disease epiphenomenon. Determining the primary causes of disease are even more problematic when considering that a succinct timeline in which a normal aging brain develops AD-like changes due to a single cause may not be appropriate, as increasing lines of evidence indicate that multiple factors likely contribute to the clinical manifestation of AD. Implications for therapeutic strategies are dramatically affected by viewing AD as a multi-factorial disease state, one specific treatment may not be able to prevent or reverse AD if this is indeed the case. In this regard, the current focus on individual therapeutic targets may prove to be ineffective in the successful treatment of AD; however, if taken in combination, these singular therapies may likely result in the global suppression of AD. In this review, the scientific basis for common AD therapeutics as well as the efficacy of these treatments will be discussed. PMID- 15892630 TI - Adding semantics to gene expression profiles: new tools for drug discovery. AB - Gene expression profiles are unveiling a wealth of new potential drug targets for a wide range of diseases, offering new opportunities for drug discoveries. The emerging challenge, however, is the effective selection of the myriad of targets to identify those with the most therapeutic utility. Numerical clustering has became a commonly used method to investigate and interpret gene expression data sets but it is often inadequate to infer the genes' and proteins' role and point to candidate genes for drug development. This review illustrates how clustering methods based on semantic characteristics, such as gene ontologies, could be used to extract more knowledge from genomic data and improve drug target and discovery processes. PMID- 15892631 TI - Metals, toxicity and oxidative stress. AB - Metal-induced toxicity and carcinogenicity, with an emphasis on the generation and role of reactive oxygen and nitrogen species, is reviewed. Metal-mediated formation of free radicals causes various modifications to DNA bases, enhanced lipid peroxidation, and altered calcium and sulfhydryl homeostasis. Lipid peroxides, formed by the attack of radicals on polyunsaturated fatty acid residues of phospholipids, can further react with redox metals finally producing mutagenic and carcinogenic malondialdehyde, 4-hydroxynonenal and other exocyclic DNA adducts (etheno and/or propano adducts). Whilst iron (Fe), copper (Cu), chromium (Cr), vanadium (V) and cobalt (Co) undergo redox-cycling reactions, for a second group of metals, mercury (Hg), cadmium (Cd) and nickel (Ni), the primary route for their toxicity is depletion of glutathione and bonding to sulfhydryl groups of proteins. Arsenic (As) is thought to bind directly to critical thiols, however, other mechanisms, involving formation of hydrogen peroxide under physiological conditions, have been proposed. The unifying factor in determining toxicity and carcinogenicity for all these metals is the generation of reactive oxygen and nitrogen species. Common mechanisms involving the Fenton reaction, generation of the superoxide radical and the hydroxyl radical appear to be involved for iron, copper, chromium, vanadium and cobalt primarily associated with mitochondria, microsomes and peroxisomes. However, a recent discovery that the upper limit of "free pools" of copper is far less than a single atom per cell casts serious doubt on the in vivo role of copper in Fenton-like generation of free radicals. Nitric oxide (NO) seems to be involved in arsenite-induced DNA damage and pyrimidine excision inhibition. Various studies have confirmed that metals activate signalling pathways and the carcinogenic effect of metals has been related to activation of mainly redox-sensitive transcription factors, involving NF-kappaB, AP-1 and p53. Antioxidants (both enzymatic and non enzymatic) provide protection against deleterious metal-mediated free radical attacks. Vitamin E and melatonin can prevent the majority of metal-mediated (iron, copper, cadmium) damage both in vitro systems and in metal-loaded animals. Toxicity studies involving chromium have shown that the protective effect of vitamin E against lipid peroxidation may be associated rather with the level of non-enzymatic antioxidants than the activity of enzymatic antioxidants. However, a very recent epidemiological study has shown that a daily intake of vitamin E of more than 400 IU increases the risk of death and should be avoided. While previous studies have proposed a deleterious pro-oxidant effect of vitamin C (ascorbate) in the presence of iron (or copper), recent results have shown that even in the presence of redox-active iron (or copper) and hydrogen peroxide, ascorbate acts as an antioxidant that prevents lipid peroxidation and does not promote protein oxidation in humans in vitro. Experimental results have also shown a link between vanadium and oxidative stress in the etiology of diabetes. The impact of zinc (Zn) on the immune system, the ability of zinc to act as an antioxidant in order to reduce oxidative stress and the neuroprotective and neurodegenerative role of zinc (and copper) in the etiology of Alzheimer's disease is also discussed. This review summarizes recent findings in the metal induced formation of free radicals and the role of oxidative stress in the carcinogenicity and toxicity of metals. PMID- 15892632 TI - The Application of Photofrin II as a sensitizing agent for ionizing radiation--a new approach in tumor therapy? AB - Radiosensitizers represent an enticing concept in tumor therapy. As ionizing radiation affects both neoplastic and normal tissues, its effects are generally non-specific. The aim of applying a radiosensitizing agent is to achieve a maximum effect on tumor tissue, while minimizing the damage to normal tissues. A variety of parameters such as the oxygen supply and the state in the cell cycle, need to be taken into account when evaluating a potential radiosensitizer. Most of the previously known radiosensitizers are neither selective nor tumor specific. In this article, we review the properties and radiosensitizing potential of Photofrin II. Photofrin II is well-known as a photosensitizing agent in photodynamic therapy. In recent years, a radiosensitizing potential of the substance has been demonstrated, specifically increasing the sensitivity of solid tumor tissues, especially of radio-resistant, hypoxic tumor cells, to radiation. This radiosensitizing effect has been demonstrated both by in vitro studies and by animal experiments. Several studies with tissue cultures have demonstrated a radiosensitizing effect of Photofrin II in glioblastoma (U-373MG) and bladder cancer cell lines (RT-4). No effect was noted in colon carcinoma cell lines (HT 29). Unpublished data of additional cell lines will be mentioned in the review. Animal experiments with Lewis sarcoma and with bladder cancer have moreover demonstrated an in vivo effect of Photofrin II as a radiosensitizer. The mechanism of this radiosensitizing effect is not completely understood. In vitro data, however, support the hypothesis that the radiosensitizing action involves OH-radicals in addition to a potential impairment of repair mechanisms after sublethal damage of ionizing radiation. Moreover, early results of a phase I trial are available and document the potential feasibility of the application of Phototofrin II as a radiosensitizing agent in clinical practice. PMID- 15892633 TI - Targeting the cannabinoid CB2 receptor: mutations, modeling and development of CB2 selective ligands. AB - Since the discovery of the cannabinoid CB2 receptor in 1993, there has been a growing interest to clarify the importance of this G-protein coupled receptor (GPCR) for human physiology, and to investigate it as a possible target for current and future drug development. Several mutation studies have examined the receptor activation and structure of the receptor binding cavity. Additionally, 3D models for the CB2 receptor have been constructed to aid in perceiving important ligand-receptor interactions. In recent years, many research groups have succeeded in synthesizing new CB2 selective ligands. This review focuses on (i) important features for ligand recognition and/or receptor activation at CB2, derived from mutation and modeling studies, and (ii) recent advances in the field of CB2 selective ligands. PMID- 15892635 TI - The nuclear receptor ligand-binding domain: a family-based structure analysis. AB - Nuclear receptors (NRs) are ligand-dependent transcription factors that play a central role in various physiological processes. The pharmaceutical industry has great interest in this gene-family for the discovery of novel or improved drugs for treatment of, for example, cancer, infertility, or diabetes. The usage of three-dimensional coordinates of protein structures to analyse and predict interactions with ligands is an important aspect of this process. All NR ligand binding domains have a similar fold, which allows for comparison of the structures of their three main functional sites: the ligand-binding pocket, the cofactor-binding groove, and the dimerization interface. We performed an analysis of nearly one hundred NR ligand-binding domain structures, and identified the functionally important residues. The combined knowledge about the shape of the binding sites and the residues involved in the binding is important for drug design in two ways. First, knowledge about the location of residues that interact with a ligand in all crystal structures or in certain subfamilies assists in the design and docking of drugs. Second, similarities and differences in the residue types of the most frequent ligand- and cofactor-binding residues provide insight about potential cross-reactivity of ligands or cofactors. PMID- 15892636 TI - Non-steroidal steroid receptor modulators. AB - The last ten years much attention has been focused on the finding of non steroidal ligands for steroidal nuclear receptors for reasons such as diminishing cross-reactivity to eliminate side effect profiles, changing physicochemical properties which might cause different tissue distribution profiles and altering binding modes which influence the binding of cofactors. Compounds with a selective functionality profile are referred to as selective nuclear receptor modulators (e.g., SARMs or SPRMs). In the following paragraphs non-steroidal ligands which have full or partial agonistic activity will be described for the following receptors: PR, GR, AR, LXR and FXR. PMID- 15892637 TI - Non-steroidal subtype selective estrogens. AB - The biological effects of estrogens are thought to be mediated by two receptors referred to as ERalpha and ERbeta. In recent years significant efforts have been devoted to the design of subtype selective ligands. These ligands are valuable tools to establish the precise biological role of each of the subtypes and to develop new generations of therapeutics. The first part of this review briefly summarizes the biology behind the estrogen receptors. The second part addresses the structure-activity relationship of the subtype selective ER ligands that were reported up to now. In the third part, the current insights in the therapeutic prospects of the subtype selective estrogens will be discussed. PMID- 15892646 TI - Genetics of common forms of glycaemia with pathological impact on vascular biology: are we on the right track? AB - The common forms of abnormal glucose regulation including type 2 diabetes and impaired glucose tolerance with pathological implications on vascular biology have a complex aetiology involving multiple cross-talks between genetic influences and important environmental modifying factors. Due to complexity of the genetics and the clinical heterogeneity of these disorders it has proven difficult to apply the same methodological approaches that have recently given insights into the molecular genetics of several single-gene disorders of glucose metabolism. This review gives some reflections on the challenges posed by the current hypotheses about the genetics of the widespread forms of abnormal glucose regulation as well as on the strengths and limitations of the methodological approaches applied to unravel the genetic components of common disorders. Also, we review recent progress in relation to a model for the pathogenesis of the various stages of abnormal glucose regulation based on the concepts of thrifty genes of metabolism and pro-inflammation and genes responsible for the appearance of impaired pancreatic beta-cell function and insulin signalling under the pressure of a westernized environment. PMID- 15892647 TI - Type 2 diabetes, insulin secretion and beta-cell mass. AB - In nondiabetic subjects, insulin secretion is sufficiently increased as a compensatory adaptation to insulin resistance whereas in subjects with type 2 diabetes, the adaptation is insufficient. Evidences for the islet dysfunction in type 2 diabetes are a)impaired insulin response to various challenges such as glucose, arginine and isoproterenol, b)defective dynamic of insulin secretion resulting in preferential reduction on first phase insulin secretion and irregular oscillations of plasma insulin and c)defective conversion of proinsulin to insulin leading to elevated proinsulin to insulin ratio. In addition, recent studies have also presented evidence of a reduced beta cell mass in diabetes, caused predominantly by enhanced islet apoptosis, although this needs to be confirmed in more studies. These defects may be caused by primary beta cell defects, such as seen in the monogenic diabetes forms of MODY, or by secondary beta cell defects, caused by glucotoxicity, lipotoxicity or islet amyloid aggregation. The defects may also be secondary to defective beta cell stimulation by incretin hormones or the autonomic nerves. The appreciation of islet dysfunction as a key factor underlying the progression from an insulin resistant state into type 2 diabetes has therapeutic implications, since besides improvement of insulin sensitivity, treatment should also aim at improving the islet compensation. This may possibly be achieved by stimulating insulin secretion, supporting islet stimulating mechanisms, removing toxic beta-cell insults and inhibiting beta cell apoptosis. PMID- 15892648 TI - The fatty liver and insulin resistance. AB - Obesity is not necessary to observe insulin resistance in humans since severe insulin resistance also characterizes patients lacking subcutaneous fat such as those with HAART (highly-active antiretroviral therapy) - associated lipodystrophy. Both the obese and the lipodystrophic patients have, however, an increase in the amount of fat hidden in the liver. Liver fat content can be non invasively accurately quantified by proton magnetic resonance spectroscopy. It is closely correlated with fasting insulin and direct measures of hepatic insulin sensitivity while the amount of subcutaneous adipose tissue is not. The causes of interindividual variation in liver fat content independent of obesity are largely unknown but could involve differences in signals from adipose tissue such as in the amount of adiponectin produced and differences in fat intake. Adiponectin deficiency characterizes both lipodystrophic and obese insulin resistant individuals, and serum levels correlate with liver fat content. Liver fat content can be decreased by weight loss. In addition, treatment of both lipodystrophic and type 2 diabetic patients with PPARgamma agonists but not metformin decreases liver fat and increases adiponectin levels. Markers of liver fat such as serum alanine aminotransferase activity have been shown to predict type 2 diabetes in several studies independent of obesity. The fatty liver thus may help to explain why some but not all obese individuals are insulin resistant and why even lean individuals may be insulin resistant, and thereby at risk of developing type 2 diabetes and cardiovascular disease. PMID- 15892649 TI - Type 2 diabetes as a lipid disorder. AB - Diabetic dyslipidemia is a cluster of plasma lipid and lipoprotein abnormalities that are metabolically interrelated. The recognition that the elevation of large VLDL 1 particles initiates a sequence of events that leads to the formation of small dense LDL and HDL species has focused the assembly of VLDL particles on the spotlight as a potential culprit of dyslipidemia. Notably dyslipidemia is associated with insulin resistance, visceral obesity and liver fat content. Insulin resistance is associated with excessive flux of substrates for VLDL assembly to the liver as well as the upregulation of the machinery generating large VLDL particles in excess. The regulation of different molecular steps in this cascade of events are complex and so far poorly understood. The disordered crosstalk between adipose tissue and the liver results in an imbalance of the machinery that orchestrates the regulation of VLDL production. A number of studies indicates that adipocytokines in particular adiponectin may be a seminal player in the regulation of fat metabolism in the liver. Future discoveries hopefully will delineate the regulatory steps to allow more targeted treatment of diabetic dyslipidemia. PMID- 15892650 TI - Type 2 diabetes as an inflammatory cardiovascular disorder. AB - Type 2 diabetes carries a 2-6-fold increased risk of cardiovascular disease (CVD) and death. Indeed, the risk of major cardiovascular events in Type 2 diabetic patients without history of coronary heart disease (CHD) is equivalent to that observed in non-diabetic subjects with CHD. However, atherosclerosis may also precede the development of diabetes, suggesting that both disorders share common genetic and environmental antecedent factors ("common soil" hypothesis). One such a possible ancestor is insulin resistance which constitutes both a major feature of Type 2 diabetes and an independent risk factor for CHD. It is well documented that inflammatory processes play an important role in the causation of atherosclerotic CVD. Inflammatory mediators play a paramount role in the initiation, progression, and rupture of atherosclerotic plaques. Thus, markers of inflammation and endothelial dysfunction may provide additional information about a patient's risk of developing CVD and may become new targets for treatment. On the other hand, evidence has emerged suggesting that inflammation is also involved in the development of Type 2 diabetes. Prospective studies have demonstrated that increased levels of pro-inflammatory markers such as CRP or reduced levels of anti-inflammatory markers such as adiponectin predict the development of Type 2 diabetes. Thus, there is accumulating evidence suggesting that inflammation is the bridging link between atherosclerosis and the metabolic syndrome. Interventions by lifestyle modification or agents with anti inflammatory properties may reduce the risk of both conditions. Drugs exerting anti-inflammatory and vascular effects have future potential to be used within an array of interventions aimed at reducing the enormous cardiovascular burden associated with Type 2 diabetes. PMID- 15892651 TI - Type 2 diabetes: an atherothrombotic syndrome. AB - Insulin resistance is found in around 80-90% of subjects with older onset (type 2) diabetes and in approximately 25% of the general population. Insulin resistance prior to the development of frank type 2 diabetes and type 2 diabetes itself is associated with a significant increase in the risk of atherothrombotic disease, which is due in part to a disruption in the balance of factors regulating coagulation and fibrinolysis. Both insulin resistance and type 2 diabetes are associated with the development of endothelial dysfunction, and enhanced platelet aggregation and activation. Whilst the plasma levels of many clotting factors including fibrinogen, FVII, FVIII, FXII, FXIII b-subunit are elevated, the fibrinolytic system is relatively inhibited as a consequence of an increase in plasminogen activator inhibitor type-1 (PAI-1) levels. These changes favour the development of a hypercoagulable pro-thrombotic state, which may in turn enhance cardiovascular risk by increasing the likelihood of developing an occlusive thrombus within a coronary/cerebral artery, and/or contributing to the development of atherosclerotic lesions. This article reviews the current published evidence of the pro-thrombotic changes that occur in association with type 2 diabetes and insulin resistance, and the putative underlying mechanisms which lead to these changes. PMID- 15892652 TI - Insulin resistance in type 2 diabetes -- role of the adipokines. AB - The role of adipocytes as protein secreting cells has been known for almost 15 years. Most of these proteins have known biological activity and are called adipokines. However, only a few of the adipokines have been shown to regulate insulin sensitivity. The latter effects are direct or indirect. The adipokines regulating insulin sensitivity are tumor necrosis factor alpha, adiponectin, interleukin-6, resistin and leptin. This review examines the mechanism how these adipokines influence insulin sensitivity, how the adipocyte production of the adipokines is regulated and if genetic variance in the genes encoding for adipokines is important for the development of type 2 diabetes mellitus. PMID- 15892653 TI - Role of AMP--activated protein kinase in the control of glucose homeostasis. AB - Skeletal muscle insulin resistance is a hallmark feature of Type 2 diabetes. Physical exercise/muscle contraction elicits an insulin-independent increase in glucose transport and perturbation of this pathway may bypass defective insulin signaling. To date, the exercise-responsive signaling molecules governing glucose metabolism in skeletal muscle are largely unknown. AMP-activated protein kinase (AMPK) has been suggested as one of the exercise-responsive signaling molecules involved in glucose homeostasis and consequently it has been heavily explored as a pharmacological target for the treatment of Type 2 diabetes. AMPK exists in heterotrimeric complexes composed of a catalytic alpha-subunit and regulatory beta- and gamma-subunits. The gamma3-isoform of AMPK is expressed specifically in skeletal muscle of humans and rodents and this tissue specific expression pattern offers selectivity in AMPK action. Furthermore, mutations in the AMPK gamma3 isoform may provide protection from diet-induced insulin resistance by increasing lipid oxidation in the presence of increased lipid supply. This review highlights the current understanding of the role of the regulatory AMPK gamma3-isoform in the control of skeletal muscle metabolism. PMID- 15892654 TI - The Clinical Significance of PPAR Gamma Agonism. AB - Insulin resistance is a principal underlying defect in type 2 DM along with beta cell dysfunction, and this insulin resistance underpins many of the abnormalities associated with the metabolic syndrome. Peroxisome-proliferator-activated receptor gamma agonists (PPARgamma agonists), also known as glitazones or thiazolidinediones (TZDs) are powerful insulin sensitisers with recent evidence suggesting that they also have a potential to improve pancreatic beta-cell function. TZDs cause a major redistribution of body fat with a decrease in visceral and hepatic fat content with a resultant increase in insulin sensitivity. The glucose lowering effects of TZDs are similar to those seen with the well-established sulphonylureas and metformin. TZDs have a small reducing effect on blood pressure and have been shown to reduce microalbuminuria independent of their blood glucose lowering effect. Both TZDs in clinical practice, pioglitazone and rosiglitazone, reduce small dense LDL-cholesterol and increase HDL-cholesterol levels but pioglitazone would appear to have a more pronounced benefit on these two parameters with a greater reduction in plasma triglycerides. TZDs improved the pro-coagulant state and show benefits in improving endothelial dysfunction and reducing 'non-traditional' inflammatory cytokines and increasing adiponectin levels. The greatest benefit for the TZDs is to directly influence atherogenesis itself and the potential that these so-called pleiotrophic effects of TZDs to reduce cardiovascular events in type 2 DM will be tested when the results of outcome trials are published in the next few years. If the results are positive for the reduction in vascular end-points, then TZDs will represent a major advance in improving the prognosis of type 2 DM subjects with the metabolic syndrome. PMID- 15892655 TI - Prevention of type 2 diabetes. AB - Changes in the human environment and in human behavior and lifestyle, in conjunction with genetic susceptibility, have resulted in a dramatic increase in the incidence and prevalence of diabetes in the world. The rapid escalation of the number of people with type 2 diabetes (T2DM) and diabetes-related cardiovascular disease demands urgent action on prevention. The Finnish Diabetes Prevention Study and The Diabetes Prevention Program showed that the prevention (or delaying) of T2DM is feasible and effective. Both of these trials led to a reduction of 58% in the conversion to diabetes in subjects with impaired glucose tolerance. Compared to lifestyle changes, drug treatment in the prevention of diabetes in people at high risk for T2DM has been less beneficial. Metformin (31%) or acarbose (25%) treatment obtained only about a half of the reduction in the conversion to diabetes compared to lifestyle changes. These drugs require monitoring, and have significant side-effects. Also the effect of orlistat (37%) did not reach the effect of lifestyle modification. Results of the Troglitazone in Prevention of Diabetes study are suggestive for the prevention, but the trial was too small, and included only one ethnic group (Hispanic) and one gender (women). On the basis of the evidence available, we do not have a definite proof that T2DM is prevented in any of these trials. However, we can safely conclude that the current evidence strongly favors the notion that lifestyle changes are the primary means to tackle the epidemic of T2DM. PMID- 15892657 TI - The dual transporter inhibitor duloxetine: a review of its preclinical pharmacology, pharmacokinetic profile, and clinical results in depression. AB - Major depressive disorder (MDD) poses a significant health problem and is estimated to be the third most costly and disabling disorder in the United States. Pharmacotherapy of depression has been successful, but improvements in response rates, remission rates, side effects, compliance and faster onset of therapeutic action have become prime objectives in drug development. There is considerable support for the hypothesis that dysfunctional serotonergic or noradrenergic neurotransmission may be etiological in depressed patients. Duloxetine is a balanced and potent reuptake inhibitor of serotonin (5-HT) and norepinephrine (NE) being studied as an antidepressant medication. In this review, we highlight the preclinical pharmacology, pharmacokinetic profile, and effects of duloxetine in the pharmacotherapy of depression. Evidence for 5-HT and NE reuptake inhibition by duloxetine comes from in vitro and in vivo transporter binding and functional uptake studies. Taken together with efficacy data from in vivo microdialysis, electrophysiological and behavioral studies, it is evident that duloxetine is balanced as a dual serotonin norepinephrine uptake inhibitor in vivo. The clinical efficacy and safety of duloxetine in the treatment of MDD has been studied in 6 multicenter, randomized, double-blind, placebo-controlled trials. In these studies, duloxetine was found to be effective in the treatment of emotional/psychological and painful physical symptoms associated with depression. More importantly, duloxetine appears to have better response rates and remission from depressive symptoms, perhaps due to its ability to treat a wider range of symptoms. PMID- 15892658 TI - Brain-derived neurotrophic factor and antidepressant activity. AB - Brain-derived neurotrophic factor (BDNF) is a member of the structurally and functionally homologous neurotrophin family. It is the most widely distributed trophic factor in the brain, and participates in neuronal growth, maintenance, and use-dependent plasticity mechanisms such as long-term potentiation and learning. There are several lines of evidence supporting a role for BDNF in the treatment of depression. This paper reviews the neurotrophin hypothesis of antidepressant action, and examines our current understanding of activity dependent mechanisms of BDNF expression and function in limbic regions of the brain. Our discussion starts with the original observations of monoaminergic neurotransmitter dysfunction that served as the basis for early antidepressant drug development, and outlines evidence for neurodegeneration and functional deficits existing with chronic stress and depression. We continue with evidence that enhancement in neurotrophic support and associated augmentation in synaptic plasticity and function may form the basis for antidepressant efficacy, and serve as a current and future focus in the quest for more rapid-acting and effective medication treatments. Finally, we follow the current search for the intracellular mechanisms of antidepressant interventions that may bring the monoaminergic and neurotrophic hypotheses together, and help us to more fully understand the roles of both neurotransmitter and growth factor. Principal challenges to the neurotrophin hypothesis, and inconsistencies between clinical and preclinical research results, are also pointed out, as these also guide future experiments that will refine our understanding of treatment mechanisms. PMID- 15892659 TI - AMPA receptor potentiators as novel antidepressants. AB - Depression affects a large percentage of the general population and can produce devastating consequences to affected individuals, families and society. Although the treatment of depression has been advanced by traditional antidepressants, improvements are needed across several dimensions (e.g., overall treatment efficacy, therapeutic onset time, and side effect profile). The alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor has an allosteric modulatory site(s) for which potent positive modulators (potentiators) have been designed. These compounds produce antidepressant-like effects in animal models, increase levels of brain-derived neurotrophic factor (BDNF) and engender neurogenesis in vivo. Although these effects are also produced by traditional antidepressants, AMPA receptor potentiators appear to produce their effects through a novel mechanism. PMID- 15892660 TI - Do substance P and the NK1 receptor have a role in depression and anxiety? AB - Research on Substance P (SP) has, until recently, focused on its role in pain and inflammation. However, a report that NK(1) receptor antagonists have utility in the treatment of depression has stimulated research into the function of SP and the NK(1)receptor in anxiety and depression. The distribution of SP and the NK(1) receptor in brain areas implicated in anxiety and depression is initially reviewed. This is followed by evaluation of the preclinical data obtained for SP and NK(1) receptor antagonists in behavioral models of depression as well as the phenotype of genetically modified animals lacking the genes encoding for the NK(1) receptor or for SP. The weight of the evidence supports antidepressant and anxiolytic activity of NK(1) receptor antagonists. However, many of the studies do not control for nonspecific effects of the compounds, and when enantiomers that lack activity at the NK(1) receptor are included, the results, in some cases, suggest that blockade of NK(1) receptors does not account for the observed behavioral activity. Finally, clinical studies in depressed patients assessing SP levels in plasma and cerebrospinal fluid as well as the effect of NK(1) receptor antagonists are reviewed. The clinical studies are a mixture of positive, failed and negative studies on the antidepressant activity of NK(1) receptor antagonists, not unlike the early clinical results obtained with selective serotonin reuptake inhibitors. PMID- 15892661 TI - Non-peptide vasopressin V1b receptor antagonists as potential drugs for the treatment of stress-related disorders. AB - Since vasopressin has been shown to be critical for adaptation of the hypothalamo pituitary-adrenal axis during stress through its ability to potentiate the stimulatory effect of CRF, it has been hypothesized that this peptide may provide a good opportunity for pharmacological treatment of stress-related disorders. The availability of the first orally active non-peptide V(1b) receptor antagonist, SSR149415, opened a new era for examining the role of vasopressin in animal models of anxiety and depression. In rats, SSR149415 blocked several endocrine (i.e. ACTH release), neurochemical (i.e. noradrenaline release) and autonomic (i.e. hyperthermia) responses following various stress exposures. Moreover, the drug was able to attenuate some but not all stress-related behaviors in rodents. While the antidepressant-like activity of the compound was comparable to that of reference antidepressants, the overall profile displayed in anxiety tests was different from that of classical anxiolytics, such as benzodiazepines. These latter were highly effective and reliably produced robust effects in most anxiety tests, while SSR149415 showed clear-cut effects only in particularly stressful situations. Experiments with mice or hamsters indicated that V(1b) receptor blockade is associated with reduced aggressiveness, suggesting that SSR149415 could prove useful for treating aggressive behavior. It is important to note that SSR149415 is devoid of adverse effects on motor functions or cognitive processes, and it did not produce tolerance to its anxiolytic- or antidepressant-like activity. Altogether, these findings suggest that V(1b) receptor antagonists represent a promising alternative to agents currently used for the treatment of depression and some forms of anxiety disorders. PMID- 15892662 TI - Specific therapy based on the genotype and cellular mechanism in inherited cardiac arrhythmias. Long QT syndrome and Brugada syndrome. AB - Seven forms of congenital long QT syndrome (LQTS) caused by mutations in ion channel genes have been identified. Genotype-phenotype correlation in clinical and experimental studies involving arterially-perfused canine left ventricular wedges suggest that beta-blockers are protective in LQT1, less so in LQT2, but not protective in LQT3. A class IB sodium channel blocker, mexiletine, is most effective in abbreviating QT interval in LQT3, but effectively reduces transmural dispersion of repolarization (TDR) and prevents the development of Torsade de Pointes (TdP) in all 3 models, suggesting its potential as an adjunctive therapy in LQT1 and LQT2. High concentrations of intravenous nicorandil, a potassium channel opener, have been shown to be capable of decreasing QT and TDR, and preventing TdP in LQT1 and LQT2 but not in LQT3. The calcium channel blocker, verapamil, has also been suggested as adjunctive therapy for LQT1, LQT2 and possibly LQT3. Experimental data using right ventricular wedge preparations suggest that a prominent transient outward current (I(to))-mediated action potential (AP) notch and a loss of AP dome in epicardium, but not in endocardium, give rise to a transmural voltage gradient, resulting in ST segment elevation and the induction of ventricular fibrillation (VF), characteristics of the Brugada syndrome. Since the maintenance of the AP dome is determined by the balance of currents active at the end of phase 1 of the AP, any intervention that reduces the outward current or boosts inward current at the end of phase 1 may normalize the ST segment elevation and suppress VF. Such interventions are candidates for pharmacological therapy of the Brugada syndrome. The infusion of isoproterenol, a beta-adrenergic stimulant, strongly augments L-type calcium current (I(Ca-L)), and is the first choice for suppressing electrical storms associated with Brugada syndrome. Quinidine, by virtue of its actions to block I(to), has been proposed as adjunctive therapy, with an implantable cardioverter defibrillator as backup. Oral denopamine, atropine or cilostazol all increase ICa-L, and for this reason may be effective in reducing episodes of VF. PMID- 15892663 TI - Novel subtype specific and universal somatostatin analogues: clinical potential and pitfalls. AB - The isolation and purification of somatostatin (SS), exactly 30 years ago, has led to the elucidation of physiologic actions of SS. This cyclic peptide is produced in the hypothalamus, throughout the central nervous system, as well as in most major peripheral organs and inhibits hormone release from the anterior pituitary gland, pancreas and the gastro-intestinal tract. The potent inhibitory actions of SS not only led to the clinical application of this peptide, but also resulted in the development of SS-analogues, among which octreotide and lanreotide are most well known and clinically used for several distinct disorders. Almost ten years ago, five different SS receptor subtypes (sst(1-5)) were identified. These receptor subtypes are variably expressed in distinct tissues and bind with varying affinity to the different SS-analogues, providing an excellent tool to unravel the (patho-) physiological function of the five sst subtypes. Following an overview upon the latest developments in SS receptor physiology and established diagnostic and therapeutic efficacy of SS-analogues in several challenging neuroendocrine disorders, this review predominantly focuses upon the latest developments of (clinically) potential novel sst subtype specific analogues as well as universal binding SS-peptides. Also, the role of potential SS antagonists is assessed. Furthermore, the most recent insights concerning targeted sst-mediated chemo- or radiotherapy are discussed, which offers new therapeutic and diagnostic opportunities for patients harbouring sst-positive neuroendocrine diseases. Finally, acknowledged side effects and possible pitfalls of the use of SS-analogues are discussed. PMID- 15892665 TI - Recent developments in the chemical biology of epothilones. AB - Epothilones A and B are naturally occurring microtubule-stabilizers, which inhibit the growth of human cancer cells in vitro at nM or even sub-nM concentrations. In contrast to paclitaxel (Taxol) epothilones are also active against different types of multidrug-resistant cancer cell lines in vitro and against multidrug-resistant tumors in vivo (epothilone B). Their attractive preclinical profile has made epothilones important lead structures in the search for improved cytotoxic anticancer drugs and epothilone B is currently undergoing phase II clinical trials. Numerous synthetic and semi-synthetic analogs have been prepared since the absolute stereochemistry of epothilone B was first disclosed in mid-1996 and their in vitro biological activity has been determined. Apart from generating a wealth of SAR information, these efforts have led to the identification of at least four compounds (in addition to epothilone B), which are currently at various stages of clinical evaluation in humans. This review is first intended to provide a summary of the basic features of the in vitro biological profile of epothilone B, with particular emphasis on recent developments in this area. A second part will outline the most relevant aspects of the epothilone SAR with regard to effects on tubulin polymerization, in vitro antiproliferative activity, and in vivo antitumor activity. This will include a brief discussion of research directed at the determination of the bioactive conformation of epothilones. In a final section, the preclinical profile of those epothilone analogs currently in clinical development will be discussed in greater detail. PMID- 15892666 TI - The epothilones and related analogues-a review of their syntheses and anti-cancer activities. AB - The macrocylic polyketide class of compounds known as the epothilones has generated substantial interest over the last few years in the areas of chemistry, biology, and medicine due to their interesting structure and, more importantly, their activity against numerous cancer cell lines, including drug-resistant, especially Taxol-resistant, cancer cell lines. To date, numerous total syntheses have been published, hundreds of epothilone analogues have been synthesized, and detailed structure activity relationship studies have been conducted. The purpose of this review is to give a brief summary of the latest advances made concerning the epothilones. Recent total or partial syntheses will be presented along with the syntheses of new epothilone analogues and their corresponding biological data. In addition, we will look at the current state of research into an economically viable method for the biosynthesis of the epothilones and related analogues. PMID- 15892667 TI - Pharmaceutical design of antimitotic agents based on combretastatins. AB - The design of novel anticancer agents based on the combretastatins, a group of antimitotic agents isolated from the bark of the South African willow tree Combretum caffrum Kuntz, is of considerable contemporary interest. Combretastatin A-4, the most active compound in the group, due to its unique dual features of antitubulin and antivascular properties, has drawn significant attention of medicinal chemists for the design of analogues as novel antitumor agents. To date, 252 references have been published since 1982 and 187 references have been published since 1998 related to combretastatins research. The 102 references related to chemistry efforts can be classified into three different categories including one-atom, two-atom, and three-atom bridgeheads as linker between two aryl rings of combretastatins. This review will particularly elucidate the rationale and strategic tactics towards the development of novel classes of antimitotic agents, based upon combretastatin A-4 as a promising lead. PMID- 15892668 TI - The chemistry and biology of antimitotic chalcones and related enone systems. AB - The development of combretastatin as an antimitotic agent has led to an enormous effort to design other tubulin-targeting agents. The intriguing discovery that combretastatin A-4 phosphate causes selective damage to tumor vasculature has stimulated even more activity in this field. This attention to tubulin binding agents and their antivasculature activity is highly likely to lead to significant clinical advances for the treatment of cancer. This review focuses on the development of ketones as tubulin-binding agents such as chalcones and related enones as surrogates of combretastatin and colchicine. PMID- 15892669 TI - Podophyllotoxin derivatives: current synthetic approaches for new anticancer agents. AB - Podophyllotoxin is an antimitotic natural product. Its inhibitory activity on cell growth led to the development of the clinically valuable anticancer agents, etoposide, teniposide and the water-soluble prodrug, etoposide phosphate. The cytotoxic mechanism of these drugs is the inhibition of topoisomerase II, unlike the lead compound which inhibits mitosis. Through extensive structure-activity relationship studies, several potential drug candidates were synthesized such as GL-331, TOP 53, NK611, and azatoxin. Recently, more complex and diverse analogues have been synthesized either to get more potent compounds or to overcome drug resistance. At the same time, a number of prodrug approaches have been tried to enhance the tumor selectivity or to increase the aqueous solubility. The prodrugs can release cytotoxic etoposide through the actions of hydrolysis, enzymes or catalytic antibodies. More sophisticated prodrug strategies have been applied in etoposide and these produced some interesting results. In this review, the current research trends in the design of new derivatives will be covered with a brief introduction of podophyllotoxin and related analogues. PMID- 15892670 TI - Improving the targeting of tubulin-binding agents: lessons from drug resistance studies. AB - Natural product drugs that target the tubulin/microtubule system remain an important component in the therapeutic arsenal to treat many types of malignancies. Agents such as the taxanes and vinca alkaloids bind to beta-tubulin and disrupt microtubule dynamics by inducing a potent mitotic block and subsequent cell death. Understanding why certain cancers do not respond to treatment or develop resistance has been the subject of numerous studies in recent years. An increasing body of evidence suggests that alterations in the drug target, such as tubulin mutations, altered microtubule dynamics, altered tubulin isotype expression, and modifications in microtubule regulatory proteins, are key mechanisms of antimicrotubule drug resistance. In addition, recent work indicates that other cytoskeletal proteins that can regulate microtubule dynamics through signaling or structural interactions may be important determinants of antimicrotubule resistance. As our understanding of drug action and resistance mechanisms has increased, we can now begin to exploit these to design strategies that overcome, or counteract resistance, hence improving the efficacy of antimicrotubule agents for the treatment of cancer. This review highlights the major areas of investigation as they relate to the tubulin/microtubule system and discusses opportunities that potentially exist for improved therapeutic benefit in the treatment of drug resistant disease. PMID- 15892672 TI - The role of COX-2 in acute pain and the use of selective COX-2 inhibitors for acute pain relief. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are the mainstay of therapy for the management of acute pain. Cyclooxygenase (COX) enzyme is of particular interest because it is the major target of NSAIDs. Although NSAIDs are remarkably effective in the management of pain and inflammation, their use is limited by several adverse effects including gastrointestinal bleeding and ulceration, impaired renal function, and inhibition of platelet aggregation. Discovery of a second cyclooxygenase, COX-2, led to the hypothesis that NSAID side effects could be decreased, as the inhibition of COX-2 is more directly implicated in ameliorating inflammation while the inhibition of COX-1 is related to adverse effects in the GI tract. This stimulated the development of selective COX-2 inhibitors (coxibs) that are better tolerated than nonselective NSAIDs but comparable in analgesic efficacy. This article provides an overview on the therapeutic use of selective COX-2 inhibitors for relief of acute pain, largely based on clinical trials in patients undergoing the surgical removal of impacted third molars, with focus on analgesic efficacy and the potential safety associated with their use compared to dual COX-1/COX-2 inhibitors. PMID- 15892673 TI - The effects of non-steroidal anti-inflammatory drugs (selective and non selective) on the treatment of periodontal diseases. AB - The objective was to review the literature on the effects of selective and non selective non-steroidal anti-inflammatory drugs (NSAIDs) on the treatment of periodontal diseases. A search of MEDLINE was conducted and articles published in English until December 2003 were included. The results from in vitro and animal experiments as well as from human clinical trials are presented. Non-selective cyclooxygenase-1 (COX-1) inhibitors used in periodontal research include compounds such as aspirin, flurbiprofen, ibuprofen, naproxen and piroxicam. Selective cyclooxygenase-2 (COX-2) inhibitors represent a new group of pharmaceutical products termed "coxibs" that include meloxicam, nimesulide, etodolac and celecoxib. Evidence from animal experiments and clinical trials documents that selective and non-selective NSAIDs are mainly responsible for the stabilization of periodontal conditions by reducing the rate of alveolar bone resorption. This is achieved through local inhibition of both enzymes (e.g. COX-1 and COX-2) responsible for the synthesis of arachidonic acid metabolites. Evidence shows that the effects of NSAIDs drop off rapidly after drug withdrawal. One of the major advantages of selective COX-2 inhibition is the reduction of adverse systemic effects. Although some studies present promising results, no data from long-term, multicenter prospective clinical trials are yet available for determining whether these therapeutic effects can be retained on a long-term basis. Many of these compounds, such as flurbiprofen, are readily absorbed through the gingival tissues. Therefore, the development of topical NSAIDs formulations (e.g. gels, toothpastes, rinses) with a daily application seems to be of particular interest. This may help to further reduce adverse systemic effects of non-selective NSAIDs in the long-term host modulation of periodontitis susceptible patients. PMID- 15892674 TI - The role of COX-2 in oral cancer development, and chemoprevention/ treatment of oral cancer by selective COX-2 inhibitors. AB - Oral cancer is challenging for clinicians due to its high mortality and increasing incidence rate. Cyclooxygenase-2 (COX-2) is extensively expressed in oral cancer and oral premalignant lesions and seems to be enhanced specifically in high-risk oral lesions. Mounting evidence suggests that these inhibitors may represent a promising approach for chemoprevention or treatment of oral cancer. This review reports on Medline and PubMed literature searches of published articles from 1995 to 2003. Our purpose is to provide a comprehensive examination and discussion of the potential role of COX-2 in oral cancer development and the use of COX-2 inhibitors for oral cancer chemoprevention or treatment. The data in the literature strongly indicate that COX-2 is significantly upregulated in oral cancer and premalignant lesions, and we believe that inhibition of COX-2 would suppress development of oral lesions by affecting several pathways of oral carcinogenesis. Therefore, the COX-2 inhibitors should be investigated as a new treatment, particularly new chemoprevention agents, for patients who are at high risk for developing oral cancer. PMID- 15892675 TI - Gastrointestinal effects of selective and non-selective non-steroidal anti inflammatory drugs. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most commonly prescribed group of drugs. Patients receiving NSAIDs often experience abdominal discomfort, and some of them develop serious gastrointestinal complications, such as ulceration, bleeding, perforation, or obstruction. Gastrointestinal side effects of NSAIDs are mostly attributed to cyclooxygenase (COX) inhibition resulting in reduction of prostaglandin in gastric mucosa. Topical irritant effects are also contributed to their systemic effect of prostaglandin inhibition. Anti-inflammatory effects of NSAIDs are mediated by COX-2 inhibition, while COX-1 inhibition is responsible for gastric prostaglandin inhibition. Management of gastrointestinal complications of NSAIDs is costly. In order to prevent or treat the gastrointestinal complications of NSAIDs, anti-ulcer drugs can be used concomitantly. Other alternative is the application or substitution of COX-2 selective inhibitors, which spare gastric mucosal prostaglandin synthesis and do not damage the gastric mucosa. Application of COX-2 selective inhibitors as a first line treatment for arthritic disorders may not be cost effective, if patients do not have any risk factors including advanced age, history of complicating peptic ulcer, concomitant anticoagulant and corticosteroid medication. Patients with risk factors or those developing gastrointestinal complications during the course of NSAID treatment can be treated with COX-2 selective inhibitors if necessary. PMID- 15892676 TI - Renal effects of non-steroidal anti-inflammatory drugs and selective cyclooxygenase-2 inhibitors. AB - Nonsteroidal antiinflammatory drugs (NSAID) are one of the most commonly used medications worldwide to inhibiting COX activity for the treatment of pain and inflammation. Their nephrotoxicity has been well documented. With the development and clinical implementation of new COX-2 inhibitors, the safety, including the effects on renal function and blood pressure, is attracting increasing attention. In the kidney, COX-2 is constitutively expressed and is highly regulated in response to alterations in intravascular volume. COX-2 metabolites have been implicated in mediation of renin release, regulation of sodium excretion and maintenance of renal blood flow. Similar to conventional NSAIDs, inhibition of COX-2 may cause edema and modest elevations in blood pressure in a minority of subjects. COX-2 inhibitors may also exacerbate preexisting hypertension or interfere with other antihypertensive drugs. Occasional acute renal failure has also been reported. Caution should be taken when COX-2 inhibitors are prescribed, especially in high-risk patients (including elderly and patients with volume depletion). Recently, agents with combined lipooxygenase/COX inhibition and agents that combine NSAIDs with a nitric oxide (NO) donor have been reported to reduce adverse renal effects. PMID- 15892677 TI - Highly active antiretroviral therapy: current state of the art, new agents and their pharmacological interactions useful for improving therapeutic outcome. AB - Highly active antiretroviral therapy (HAART) dramatically changed the course of HIV infection. Currently, this therapy involves the use of agents from at least two distinct classes of antivirals: a protease inhibitor (PI) in combination with two nucleoside/nucleotide reverse transcriptase inhibitors (N(t)RTIs), or a non nucleoside reverse transcriptase inhibitor (NNRTI) in combination with NRTIs. Recently, the third family of antivirals started to be used clinically, with the advent of enfuvirtide, the first fusion inhibitor (FI). Several pharmacological agents are available form these classes of antivirals, NRTIs, NNRTIs, PIs and FIs, which will be briefly reviewed here. Some more agents are in advanced clinical evaluation or have recently been approved (such as tenofovir, a NtRTI; atazanavir, a PI; tipranavir, another PI), mainly against drug-resistant viruses. Compounds inhibiting HIV integrase, the third enzyme of HIV, are also available ultimately, with several such derivatives in clinical trials (L-731, 988 and S 1360). Another approach to inhibit the growth of retroviruses, including HIV, targets the ejection of zinc ions from critical zinc finger viral proteins, which has as a consequence the inhibition of viral replication in the absence of mutations leading to drug resistance phenotypes. All steps in the process of HIV entry into the cell may be targeted by specific compounds that might be developed as novel types of antiretrovirals. Thus, inhibitors of the gp120-CD4 interaction have been detected (zintevir, FP-21399 and BMS-378806 in clinical trials). Small molecule chemokine antagonists acting as HIV entry inhibitors also were described in the last period, which interact both with the CXCR4 coreceptor (such as AMD3100; AMD3465; ALX40-4C; T22, T134 and T140), or which are antagonist of the CCR5 coreceptor (TAK-779, TAK-220, SCH-C, SCH-D, E913, AK-602 and NSC 651016 in clinical trials), together with new types of fusion inhibitors possessing the same mechanism of action as enfuvirtide (such as T1249). Compounds interacting with Tat/Tar have also been detected which inhibit HIV replication in low micromolar range (EM2487, tamacrazine, CGP 64222 or CGA 137053 among others). Unexploited viral and cellular targets (such as the maturation process-with a first potent compound available, PA-457; the cellular proteins Tsg101, APOBEC3G, or the viral ones Vif, Rev or RNase H) are also presented, together with recently emerged approaches for eradication of HIV reservoirs. A review on the pharmacology and interactions of these agents with other drugs is presented here, with emphasis on how these pharmacological interferences may improve the clinical use of antivirals, or how side effects due to these drugs may be managed better by taking them into account. PMID- 15892678 TI - Strategies for the design of potent and selective kinase inhibitors. AB - The characterisation of the human kinome in recent years has resulted in the emergence of numerous kinase drug targets in a variety of therapeutic areas. Through the elucidation of the sequence and structural composition of kinase active sites, coupled with the solution of numerous ATP competitive ligand complex structures, significant advances have been made in developing inhibitors that are highly selective. This has shown to be the case not only for kinases that are divergent in primary structure, but also for isoforms with highly conserved structure and ATP binding sites. Here we review the methods employed in the generation of selective inhibitors and describe several successful examples of the design of highly potent and selective kinase ATP competitive ligands. We also describe examples where an alternate approach to selectivity was used. These include the use of small molecules to sequester kinases in inactive conformations, and to block phospho-transferase activity by preventing substrate docking and recruitment. Substrate recruitment sites are promising from a structure based design perspective as they contain features unique to individual protein kinases. PMID- 15892679 TI - Chemical effectors in drug discovery. PMID- 15892680 TI - Target-related affinity profiling: Telik's lead discovery technology. AB - Target-Related Affinity Profiling (TRAP) is a computational drug discovery technology that is based on 'affinity fingerprints', which are molecular descriptors derived from the protein binding preferences of small molecules. The underlying concepts of TRAP are reviewed. Affinity fingerprints are compared to molecular descriptors derived from chemical structures and shown to be a useful alternative for lead discovery. The TRAP screening process is described and two example applications are presented: I. the discovery of novel inhibitors of human intestinal carboxylesterase, and II. the discovery of novel inhibitors of cyclooxygenase-1 through the use of the affinity fingerprints of known cyclooxygenase-1 inhibitors. A summary of the complementary advantages of TRAP screening technology compared to traditional approaches to drug lead discovery concludes the review. PMID- 15892681 TI - Small molecules and future regenerative medicine. AB - Recent advances in stem cell biology may make possible new approaches for the treatment of a number of diseases including cardiovascular disease, neurodegenerative disease, musculoskeletal disease, diabetes and cancer. These approaches could involve cell replacement therapy and/or drug treatment to stimulate the body's own regenerative capabilities by promoting survival, migration/homing, proliferation, and differentiation of endogenous stem/progenitor cells. However, such approaches will require identification of renewable cell sources of engraftable functional cells, an improved ability to manipulate their proliferation and differentiation, as well as a better understanding of the signaling pathways that control their fate. Cell-based phenotypic and pathway-specific screens of synthetic small molecules and natural products have historically provided useful chemical ligands to modulate and/or study complex cellular processes, and recently provided a number of small molecules that can be used to selectively regulate stem cell fate and developmental signaling pathways. Such molecules will likely provide new insights into stem cell biology, and may ultimately contribute to effective medicines for tissue repair and regeneration. PMID- 15892682 TI - Key aspects of the Novartis compound collection enhancement project for the compilation of a comprehensive chemogenomics drug discovery screening collection. AB - The NIBR (Novartis Institutes for BioMedical Research) compound collection enrichment and enhancement project integrates corporate internal combinatorial compound synthesis and external compound acquisition activities in order to build up a comprehensive screening collection for a modern drug discovery organization. The main purpose of the screening collection is to supply the Novartis drug discovery pipeline with hit-to-lead compounds for today's and the future's portfolio of drug discovery programs, and to provide tool compounds for the chemogenomics investigation of novel biological pathways and circuits. As such, it integrates designed focused and diversity-based compound sets from the synthetic and natural paradigms able to cope with druggable and currently deemed undruggable targets and molecular interaction modes. Herein, we will summarize together with new trends published in the literature, scientific challenges faced and key approaches taken at NIBR to match the chemical and biological spaces. PMID- 15892683 TI - Ligand-receptor engineering and its application towards the complementation of genetic disease and target identification. AB - In some instances, small molecules can restore function to proteins that are impaired by genetic mutations. There are now many examples where non-specific molecules or specific ligands can act as chemical chaperones to fold proteins or stabilize folded proteins harboring genetic mutations. In contrast a few recent examples have shown that functionally impaired proteins that are stably folded can be "functionally rescued" by appropriate small molecules. Compounds that can rescue functionally impaired proteins may provide new strategies for the treatment of genetic diseases such as rickets and resistance to thyroid hormone (RTH). In addition mutant-complementing analogs and substrates that act exclusively on mutant proteins are providing important tools for the study of complex biological systems that are controlled by molecules that have multiple cellular targets. PMID- 15892684 TI - Components of successful lead generation. AB - This review aims to give an overview of current good practice in the prosecution of Lead Generation. It will assess experiences across the field as judged from the contents of the limited number of peer-review disclosures to date. It will also rely heavily on the experiences of the authors from many campaigns within this organisation. Its focus will be on the assembly of an appropriate compound collection for application in High Throughput Screening (HTS), the prosecution of HTS, the profiling of HTS output and, lastly the Hit-to-Lead optimisation of selected HTS output. Excluded from the scope are detailed aspects of library design [1], parallel synthesis [2], virtual library applications [3], virtual screening [4] and fragment screening [5] approaches, all of which have been the subject of recent reviews. PMID- 15892685 TI - Synthesis and pharmacology of new camptothecin drugs. AB - Camptothecin (CPT) drugs exhibit antineoplastic activity against colorectal, breast, lung and ovarian cancers. This review briefly summarizes the pharmacology of CPT drugs, examines four strategies and methods for the synthesis of camptothecins, and finally discusses homocamptothecins and silatecans, two new classes of CPT analog. PMID- 15892686 TI - Structure and biological activity of heparinoid. AB - Heparin is a biogenic anionic charged sulfated polysaccharide that has a range of desired activities including inhibition of tumor metastasis and inhibition of restenosis. However, its clinical use is limited to treating blood-clotting disorders. Anionic macromolecules called heparinoids have been investigated with the objective of developing heparin-like molecules with reduced anti-coagulant activity and selective anti-metastasis and anti-restenosis activity. This mini review summarizes the synthesis and biological activity of the main synthetic heparinoids reported in the past three decades. PMID- 15892687 TI - Antitumour quinones. AB - Quinones still comprise one of the largest classes of antitumour agents. For example, the anthracycline antibiotics are among the most utilised anticancer agents ever developed. Many other quinones were tested for their anticancer activity. Though there are general and well-established mechanisms for quinone toxicity, the exact contribution of the quinone moiety to the cytotoxic effect remains frequently uncertain. However, DNA represents the main target for quinoid antitumour agents and most of them belong to the groups of DNA intercalating and/or alkylating agents. But also other cellular structures such as heat shock protein 90 or telomerase have been identified as targets for quinoid compounds. PMID- 15892688 TI - Advancements in the anti-diabetes chemotherapeutics based on amino acids, peptides, and peptidomimetics. AB - Diabetes Mellitus (DM) is a highly prevalent chronic disease. Recent years have witnessed development of many new oral drugs; novel insulin analogs and their delivery systems for the treatment of patients with either type-1 or type-2 DM. The impetus for developing new antidiabetic drugs comes from the unmet need of pharmacological tools that allow diabetic patients to achieve recommended glucose control targets by precise, safe and effective ways. The number of people afflicted with DM worldwide has increased considerably in recent years and is projected to increase dramatically over the next decades. In the recent times, design and synthesis of bioactive peptides and peptidomimetics has undergone a paradigm shift. Non-proteinogenic amino acids, peptides and peptidomimetics are emerging as novel drug candidates for the treatment of various diseases and/or disorders. This review mainly discusses the advancements in the usage of unnatural amino acids, peptides and peptidomimetics as potential therapeutic agents for the treatment of DM. PMID- 15892689 TI - Human acetylcholinesterase inhibitors: electronic-topological and neural network approaches to the structure-activity relationships study. AB - The Electronic-Topological (ETM) and Neural Network methods were applied to the study of the "structure-acetylcholinesterase (AChE) inhibitor activity" relationships for a series of physostigmine and N-benzylpiperidine derivatives. Molecular fragments specific for active compounds and breaks of activity were calculated for human AChE by applying the ETM and Neural Network methods. Requirements necessary for a compound to be active were formulated; they are the result of detailed analysis of all compounds under study. A comparative study of the activity features found for human AChE was performed. PMID- 15892690 TI - Development in lipid drugs. AB - Lipopeptide lipid moieties induce dendritic cell (DC) internalization and epitopes are recognized by MHC, the major histocompatibility complex. HIV-1 (human immunodeficiency virus type 1) lipopeptide vaccine candidate elicits immune responses, and sustains HIV control after highly active antiretroviral therapy (HAART). Mp- and Dp-MART (anti-melanoma lipopeptides) induce strong CTL (cytolytic T lymphocyte) response. New BGTC, BGDA, TGKC lipoplexes mediate gene delivery, e.g., into mouse pancreatic tumor nodules. Triterpene glycyrrhizic acid (GL) inhibits SARS-CoV (severe acute respiratory syndrome associated coronavirus) replication. Compared to CDV (cidofovir), CDV ether lipid esters have enhanced activity against vaccinia (VV) and cowpox (CV) viruses in vitro. Oral treatment of VV and CV infected mice with CDV ether lipid esters, as effective as i.p. CDV, may be useful against orthopoxvirus infections in humans. PMID- 15892691 TI - The chemotherapy of chagas' disease: an overview. AB - The review presents: a) a brief description of the disease; b) a summary of the most important metabolic targets so far identified in Trypanosome cruzi (T. cruzi) along with corresponding inhibitor compounds; c) the current state of knowledge on the trypanothione reductase system of trypanosomatids with reference to oxidative stress defenses; d) detailed discussions on T. cruzi trypanothione reductase inhibitors such as nitrofuranes, naphthoquinones and phenothiazines. As yet, the chemotherapy of Chagas' disease remains an unsolved problem. Further search for new drugs must continue by means of nucleating existing chemotherapy efforts. PMID- 15892692 TI - In situ localization of sulphur in the thioautotrophic symbiotic model Lucina pectinata (Gmelin, 1791) by cryo-EFTEM microanalysis. AB - BACKGROUND INFORMATION: Lucina pectinata is a large tropical lucinid known to harbour sulphide-oxidizing bacteria in specialized gill cells. Conventional TEM (transmission electron microscopy) has shown that bacteriocytes also harbour visibly 'empty' vesicles whose chemical content remains, to date, only roughly determined. RESULTS: In the present study, L. pectinata gill tissues were cryo fixed as fast as possible by performing high-pressure freezing before a freeze substitution process and finally performing a cryo-embedding in Lowicryl. Ultrathin sections were then used for a cryo-EFTEM (where EFTEM stands for energy filtered TEM) microanalysis. Results show that bacteriocytes within the gill tissues contain elemental sulphur in small vesicles produced by the host itself. In instances of sporadic depletion of sulphur in the environment, such structures may act as energy sources for bacterial endosymbionts. CONCLUSIONS: The cryo EFTEM techniques represent (i) the only method used to date to locate and preserve sulphur at the cellular level and (ii) a powerful tool for sulphur metabolism analysis in thioautotrophic symbiont relationships. PMID- 15892693 TI - Determinants of AQP6 trafficking to intracellular sites versus the plasma membrane in transfected mammalian cells. AB - BACKGROUND INFORMATION: Most AQPs (aquaporins) function at the plasma membrane, however AQP6 is exclusively localized to membranes of intracellular vesicles in acid-secreting type-A intercalated cells of renal collecting ducts. The intracellular distribution indicates that AQP6 has a function distinct from trans epithelial water movement. RESULTS: We show by mutational analyses and immunofluorescence that the N-terminus of AQP6 is a determinant for its intracellular localization. Presence or absence at the plasma membrane of AQP6 constructs was confirmed by electrophysiological methods. Addition of a GFP (green fluorescent protein) or a HA (haemagglutinin) epitope tag (GFP-AQP6 or HA AQP6) to the N-terminus of AQP6, directed AQP6 to the plasma membranes of transfected Madin-Darby canine kidney cells. In contrast, addition of a GFP tag to the C-terminus (AQP6-GFP) caused the protein to remain intracellular, similar to untagged wild-type AQP6. Replacement of the N-terminus of AQP6 by that of AQP1 also directed AQP6 to the plasma membranes, whereas the N-terminus of AQP6 retained AQP1 in cytosolic sites. CONCLUSION: Our results suggest that the N terminus of AQP6 is critical for trafficking of the protein to the intracellular sites. Moreover, our studies provide an approach for future identification of proteins involved in vesicle sorting in the acid-secreting type-A intercalated cells. PMID- 15892694 TI - The vibrio that sheds light. PMID- 15892695 TI - Persistence of vibrios in marine bivalves: the role of interactions with haemolymph components. AB - Marine bivalves are widespread in coastal environments and, due to their filter feeding habit, they can accumulate large numbers of bacteria thus acting as passive carriers of human pathogens. Bivalves possess both humoral and cellular defence mechanisms that operate in a co-ordinated way to kill and eliminate infecting bacteria. Vibrio species are very abundant in coastal waters and are commonly isolated from edible bivalves tissues where they can persist after depuration processes in controlled waters. Such observations indicate that vibrios are regular components of bivalve microflora and that the molluscs can represent an important ecological niche for these bacteria. Here we tried to summarize data on the interactions between vibrios and bivalve haemolymph; the available evidence supports the hypothesis that persistence of bacteria in bivalve tissues depends, at least in part, on their sensitivity to the bactericidal activity of the haemolymph. Results obtained with an in vitro model of Vibrio cholerae challenged against Mytilus galloprovincialis haemocytes indicate that bacterial surface components, soluble haemolymph factors and the signalling pathways of the haemocyte host are involved in determining the result of vibrio-haemolymph interactions. PMID- 15892696 TI - Evidence for in situ crude oil biodegradation after the Prestige oil spill. AB - In November 2002, the oil tanker Prestige sank off the Spanish coast after releasing approximately 17,000 tones of heavy fuel, coating several hundred kilometers of coastline in oil sludge. In December 2002 and February 2003, samples were collected from the shore of the Galician coast to analyse the indigenous population ability to carry out crude oil degradation in situ. Carbon isotopic ratio of the dissolved inorganic carbon (DIC) in seawater samples was used as a rapid method to directly assess activity of microbes on the oil components. 12CO2/13CO2 ratio in samples from certain locations along the coast revealed degradation of a very delta13C-negative source such as the Prestige crude oil (-30.6 per thousand). Putative biodegradation processes taking place at areas with high income of fresh seawater could not be detected with this technique. Laboratory-scale biostimulation processes carried out in samples with the highest oil biodegradation activity showed that N/P deficiency in seawater is a limiting factor for crude oil degradation. The most probable number (MPN) of crude oil component degraders was estimated for several aromatic compounds (naphthalene, anthracene, phenanthrene, pyrene) and for undecane. Our results clearly show that bacteria present in the contaminated water are readily able to transform components of the crude oil into inorganic carbon. PMID- 15892697 TI - Impact of lime, nitrogen and plant species on fungal community structure in grassland microcosms. AB - A microcosm-based approach was used to study impacts of plant and chemical factors on the fungal community structure of an upland acidic grassland soil. Seven plant species typical of both unimproved and fertilized grasslands were either left unamended or treated with lime, nitrogen or lime plus nitrogen. Fungal community structure was assessed by a molecular approach, fungal automated ribosomal intergenic spacer analysis (FARISA), while fungal biomass was estimated by measuring soil ergosterol content. Addition of nitrogen (with or without lime) had the largest effect, decreasing soil pH, fungal biomass and fungal ribotype number, but there was little corresponding change in fungal community structure. Although different plant species were associated with some changes in fungal biomass, this did not result in significant differences in fungal community structure between plant species. Addition of lime alone caused no changes in fungal biomass, ribotype number or community structure. Overall, fungal community structure appeared to be more significantly affected through interactions between plant species and chemical treatments, as opposed to being directly affected by changes in individual improvement factors. These results were in contrast to those found for the bacterial communities of the same soils, which changed substantially in response to chemical (lime and nitrogen) additions. PMID- 15892698 TI - Transcriptional profiling of the hyperthermophilic methanarchaeon Methanococcus jannaschii in response to lethal heat and non-lethal cold shock. AB - Temperature shock of the hyperthermophilic methanarchaeon Methanococcus jannaschii from its optimal growth temperature of 85 degrees C to 65 degrees C and 95 degrees C resulted in different transcriptional responses characteristic of both the direction of shock (heat or cold shock) and whether the shock was lethal. Specific outcomes of lethal heat shock to 95 degrees C included upregulation of genes encoding chaperones, and downregulation of genes encoding subunits of the H+ transporting ATP synthase. A gene encoding an alpha subunit of a putative prefoldin was also upregulated, which may comprise a novel element in the protein processing pathway in M. jannaschii. Very different responses were observed upon cold shock to 65 degrees C. These included upregulation of a gene encoding an RNA helicase and other genes involved in transcription and translation, and upregulation of genes coding for proteases and transport proteins. Also upregulated was a gene that codes for an 18 kDa FKBP-type PPIase, which may facilitate protein folding at low temperatures. Transcriptional profiling also revealed several hypothetical proteins that respond to temperature stress conditions. PMID- 15892699 TI - Ecological implications of the emergence of non-toxic subcultures from toxic Microcystis strains. AB - Two toxic, microcystin-producing, Microcystis sp. strains KLL MG-K and KLL MB-K were isolated as single colonies on agar plates from Lake Kinneret, Israel. Two non-toxic subcultures, MG-J and MB-J spontaneously succeeded the toxic ones under laboratory conditions. Southern analyses showed that MG-J and MB-J are lacking at least 34 kb of the mcy region, encoding the microcystin synthetase. Analyses of the 16S rRNA genes, the intergenic spacer region between cpcB and cpcA and the patterns of the polymerase chain reaction products of randomly amplified polymorphic DNA and highly iterated palindrome, and presence of mobile DNA elements did not allow unequivocal distinction between toxic and non-toxic subcultures. Laboratory and field experiments indicated an advantage of the toxic strain over its non-toxic successor. When grown separated by a membrane, which allowed passage of the media but not the cells, MG-K severely inhibited the growth of MG-J. Furthermore, when MG strains were placed in dialysis bags in Lake Kinneret during the season in which Microcystis is often observed, cells of MG-J lysed, whereas MG-K survived. Mechanisms whereby the non-toxic subcultures emerged and prevailed over the corresponding toxic ones under laboratory conditions, as well as a possible role of microcystin under natural conditions, are discussed. PMID- 15892700 TI - Physiological and molecular characterization of a microbial community established in unsaturated, petroleum-contaminated soil. AB - The microbial communities established in soil samples from an unsaturated, petroleum-contaminated zone and from an adjacent uncontaminated site were characterized by physiological and molecular approaches. Possible electron acceptors such as sulfate and nitrate had been completely depleted in these soil samples. Slurries of these soil samples were incubated in bottles in the presence of hydrocarbon indicators (benzene, toluene, xylene and decane), and the degradation of these compounds was examined. Supplementation with electron acceptors stimulated hydrocarbon degradation, although the stimulatory effect was small in the contaminated soil. The initial degradation rates in the contaminated soil under fermentative/methanogenic conditions were comparable to those under aerobic conditions. The microbial populations in the original soil samples were analysed by cloning and sequencing of polymerase chain reaction (PCR)-amplified bacterial and archaeal 16S rRNA gene fragments, showing that the sequences retrieved from these soils were substantially different. For instance, Epsilonproteobacteria, Gammaproteobacteria, Crenarchaeota and Methanosarcinales could only be detected at significant levels in the contaminated soil. Denaturing gradient gel electrophoresis (DGGE) analyses of 16S rRNA gene fragments amplified by PCR from the incubated soil-slurry samples showed that supplementation of the electron acceptors resulted in a shift in the major populations, while the DGGE profiles after incubating the contaminated soil under the fermentative/methanogenic conditions were not substantially changed. These results suggest that petroleum contamination of the unsaturated zone resulted in the establishment of a fermentative/methanogenic community with substantial hydrocarbon-degrading potential. PMID- 15892701 TI - Assemblages of ericoid mycorrhizal and other root-associated fungi from Epacris pulchella (Ericaceae) as determined by culturing and direct DNA extraction from roots. AB - Ericoid mycorrhizal fungal endophytes form mycorrhizal associations with Ericaceae plant taxa and are regarded as essential to the ecological fitness of the plants in extremely nutrient-poor soils worldwide. We isolated fungi from roots of Epacris pulchella (Ericaceae) in a south-eastern Australian sclerophyll forest and compared rDNA internal transcribed spacer (ITS) restriction fragment length polymorphisms (RFLPs) and sequences for the cultured isolate assemblage with fungi identified in DNA extracted directly from the same root systems by cloning or denaturing gradient gel electrophoresis (DGGE). The most abundant RFLP types in the cultured isolate assemblage were identified as putative ericoid mycorrhizal ascomycete endophytes, and these also represented the most abundant RFLP types in the cloned assemblage and the most intense bands in DGGE profiles. Each method identified unique taxa, notably putative basidiomycetes in the DNA extracted directly from E. pulchella roots. However, the relative abundance of these was low. PMID- 15892702 TI - Stable isotope probing analysis of the influence of liming on root exudate utilization by soil microorganisms. AB - Rhizosphere microorganisms play an important role in soil carbon flow, through turnover of root exudates, but there is little information on which organisms are actively involved or on the influence of environmental conditions on active communities. In this study, a 13CO2 pulse labelling field experiment was performed in an upland grassland soil, followed by RNA-stable isotope probing (SIP) analysis, to determine the effect of liming on the structure of the rhizosphere microbial community metabolizing root exudates. The lower limit of detection for SIP was determined in soil samples inoculated with a range of concentrations of 13C-labelled Pseudomonas fluorescens and was found to lie between 10(5) and 10(6) cells per gram of soil. The technique was capable of detecting microbial communities actively assimilating root exudates derived from recent photo-assimilate in the field. Denaturing gradient gel electrophoresis (DGGE) profiles of bacteria, archaea and fungi derived from fractions obtained from caesium trifluoroacetate (CsTFA) density gradient ultracentrifugation indicated that active communities in limed soils were more complex than those in unlimed soils and were more active in utilization of recently exuded 13C compounds. In limed soils, the majority of the community detected by standard RNA DGGE analysis appeared to be utilizing root exudates. In unlimed soils, DGGE profiles from 12C and 13C RNA fractions differed, suggesting that a proportion of the active community was utilizing other sources of organic carbon. These differences may reflect differences in the amount of root exudation under the different conditions. PMID- 15892703 TI - Evidence for the presence of a CmuA methyltransferase pathway in novel marine methyl halide-oxidizing bacteria. AB - Marine bacteria that oxidized methyl bromide and methyl chloride were enriched and isolated from seawater samples. Six methyl halide-oxidizing enrichments were established from which 13 isolates that grew on methyl bromide and methyl chloride as sole sources of carbon and energy were isolated and maintained. All isolates belonged to three different clades in the Roseobacter group of the alpha subdivision of the Proteobacteria and were distinct from Leisingera methylohalidivorans, the only other identified marine bacterium that grows on methyl bromide as sole source of carbon and energy. Genes encoding the methyltransferase/corrinoid-binding protein CmuA, which is responsible for the initial step of methyl chloride oxidation in terrestrial methyl halide-oxidizing bacteria, were detected in enrichments and some of the novel marine strains. Gene clusters containing cmuA and other genes implicated in the metabolism of methyl halides were cloned from two of the isolates. Expression of CmuA during growth on methyl halides was demonstrated by analysis of polypeptides expressed during growth on methyl halides by SDS-PAGE and mass spectrometry in two isolates representing two of the three clades. These findings indicate that certain marine methyl halide degrading bacteria from the Roseobacter group contain a methyltransferase pathway for oxidation of methyl bromide that may be similar to that responsible for methyl chloride oxidation in Methylobacterium chloromethanicum. This pathway therefore potentially contributes to cycling of methyl halides in both terrestrial and marine environments. PMID- 15892704 TI - Ecotype diversity in the marine picoeukaryote Ostreococcus (Chlorophyta, Prasinophyceae). AB - The importance of the cyanobacteria Prochlorococcus and Synechococcus in marine ecosystems in terms of abundance and primary production can be partially explained by ecotypic differentiation. Despite the dominance of eukaryotes within photosynthetic picoplankton in many areas a similar differentiation has never been evidenced for these organisms. Here we report distinct genetic [rDNA 18S and internal transcribed spacer (ITS) sequencing], karyotypic (pulsed-field gel electrophoresis), phenotypic (pigment composition) and physiological (light limited growth rates) traits in 12 Ostreococcus strains (Prasinophyceae) isolated from various marine environments and depths, which suggest that the concept of ecotype could also be valid for eukaryotes. Internal transcribed spacer phylogeny grouped together four deep strains isolated between 90 m and 120 m depth from different geographical origins. Three deep strains displayed larger chromosomal bands, different chromosome hybridization patterns, and an additional chlorophyll (chl) c-like pigment. Furthermore, growth rates of deep strains show severe photo inhibition at high light intensities, while surface strains do not grow at the lowest light intensities. These features strongly suggest distinct adaptation to environmental conditions encountered at surface and the bottom of the oceanic euphotic zone, reminiscent of that described in prokaryotes. PMID- 15892705 TI - Marine diatom species harbour distinct bacterial communities. AB - We examined bacterial dynamics in batch cultures of two axenic marine diatoms (Thalassiosira rotula and Skeletonema costatum). The axenic diatoms were inoculated with natural bacterial assemblages and monitored by 4,6-diamidino-2 phenolindole (DAPI) counts, denaturing gradient gel electrophoresis (DGGE) with subsequent analysis of excised, sequenced 16S rRNA gene fragments, and fluorescence in situ hybridization (FISH) with group-specific 16S rRNA oligonucleotide probes. Our results show that algal growth exhibited pronounced differences in axenic treatments and when bacteria were present. Bacterial abundance and community structure greatly depended on species, growth and physiological status of even closely related algae. Free-living and phytoplankton associated bacteria were very different from each other and were dominated by distinct phylogenetic groups. The diatom-associated bacteria mainly belonged to the Flavobacteria-Sphingobacteria group of the Bacteroidetes phylum whereas free living bacteria, which were rather similar in both cultures, comprised mainly of members of the Roseobacter group of alpha-Proteobacteria. Presence and disappearance of specific bacteria during algal growth indicated pronounced differences in environmental conditions over time and selection of bacteria highly adapted to the changing conditions. Tight interactions between marine bacteria and diatoms appear to be important for the decomposition of organic matter and nutrient cycling in the sea. PMID- 15892706 TI - Moderately thermophilic Myxobacteria: novel potential for the production of natural products isolation and characterization. AB - Novel moderately thermophilic Myxobacteria from both known suborders (Cystobacterineae and Sorangineae) were isolated from soil samples of semiarid and warm climates. The addition of the anthelmintic and amoebizidal agent levamisole was a new and crucial improvement for the fast isolation of overall 37 strains, which grew very fast at temperatures of 42-44 degrees C. When the 16S rDNAs were compared with GenBank data of common Myxobacteria, identities were 98 99%, thus not reflecting the physiological differences. Similar to the Myxobacteria described so far, the new isolates are multiresistant against a variety of antibiotics and are producers of typical myxobacterial secondary metabolites. Analysis of our previous strain collection isolated from soil samples taken worldwide revealed a more or less uniform distribution of strains which synthesize specific metabolites. Therefore these moderately thermophilic Myxobacteria, which grow 2-3 times faster, have the potential to replace the slow growing isolates and provide a means for fast and cost-saving production of myxobacterial metabolites in the future. PMID- 15892707 TI - Modelling viral impact on bacterioplankton in the North Sea using artificial neural networks. AB - The temporal variability of the viral impact on bacterioplankton during the summer-winter transition in the North Sea was determined and artificial neural networks (ANNs) were developed to predict viral production and the frequency of infected bacterial cells (FIC). Viral production and FIC were estimated using a virus-dilution approach during four cruises in the southern North Sea between July and December 2000 and an additional cruise in June 2001. Supplementary data such as bacterial production, and bacterial and viral abundance were collected to relate changes in FIC and viral production to the dynamics of other biotic parameters. Average viral abundance varied between 4.4 x 10(6) ml(-1) in December and 29.8 x 10(6) ml(-1) in July. Over the seasonal cycle, viral abundance correlated best with bacterial production. Average bacterial abundance varied between 0.5 x 10(6) ml(-1) in December and 1.3 x 10(6) ml(-1) in July. Monthly average values of FIC ranged from 9% in September to 39% in June and the average viral production from 11 x 10(4) ml(-1) h(-1) in December to 35 x 10(4) ml(-1) h( 1) in July. The data set was used to develop ANN-based models of viral production and FIC. Viral production was modelled best using sampling time, and bacterial and viral abundance as input parameters to an ANN with two hidden neurons. Modelling of FIC was performed using bacterial production as an additional input parameter for an ANN with three hidden neurons. The models can be used to simulate viral production and FIC based on regularly recorded and easily obtainable parameters such as bacterial production, bacterial and viral abundance. PMID- 15892708 TI - Characterization of starvation-induced dispersion in Pseudomonas putida biofilms. AB - The biofilm lifestyle, where microbial cells are aggregated because of expression of cell-to-cell interconnecting compounds, is believed to be of paramount importance to microbes in the environment. Because microbes must be able to alternate between sessile and planktonic states, it is anticipated that they must be able to regulate their ability to form biofilm and to dissolve biofilm. We present an investigation of a biofilm dissolution process occurring in flow chamber-grown Pseudomonas putida biofilms. Local starvation-induced biofilm dissolution appears to be an integrated part of P. putida biofilm development that causes characteristic structural rearrangements. Rapid global dissolution of entire P. putida biofilms was shown to occur in response to carbon starvation. Genetic analysis suggested that the adjacent P. putida genes PP0164 and PP0165 play a role in P. putida biofilm formation and dissolution. PP0164 encodes a putative periplasmic protein of previously unknown function, and PP0164 mutant bacteria are sticky, and unable to reduce their adhesiveness and dissolve their biofilm in response to carbon starvation. PP0165 encodes a putative transmembrane protein containing GGDEF and EAL domains, and PP0165 mutant bacteria are unable to increase their adhesiveness and form biofilm. We suggest that the PP0164 and PP0165 proteins are involved in the regulation of the adhesiveness of the bacteria; the PP0165 protein through c-di-GMP signalling, and the PP0164 protein as a transducer of the signal. PMID- 15892710 TI - Editorial: melanomas and Wnts. PMID- 15892711 TI - Adhesion, migration and communication in melanocytes and melanoma. AB - Under normal conditions, homeostasis determines whether a cell remains quiescent, proliferates, differentiates, or undergoes apoptosis. In this state of homeostasis, keratinocytes control melanocyte growth and behaviour through a complex system of paracrine growth factors and cell-cell adhesion molecules. Alteration of this delicate homeostatic balance and can lead to altered expression of cell-cell adhesion and cell communication molecules and to the development of melanoma. Melanoma cells escape from this control by keratinocytes through three major mechanisms: (1) down-regulation of receptors important for communication with keratinocytes such as E-cadherin, P-cadherin, desmoglein and connexins, which is achieved through growth factors produced by fibroblasts or keratinocytes; (2) up-regulation of receptors and signalling molecules not found on melanocytes but important for melanoma-melanoma and melanoma-fibroblast interactions such as N-cadherin, Mel-CAM, and zonula occludens protein-1 (ZO-1); (3) loss of anchorage to the basement membrane because of an altered expression of the extracellular-matrix binding integrin family. In the current review, we describe the alterations in cell-cell adhesion and communication associated with melanoma development and progression, and discuss how a greater understanding of these processes may aid the future therapy of this disease. PMID- 15892712 TI - Histone deacetylase inhibitors and malignant melanoma. AB - The search for antimelanoma agents acting by terminal differentiation via the pigmentation pathway has so far been unsuccessful, in part because of tumor heterogeneity and loss of function of pigmentation genes. Some differentiation agents, however, have emerged as inhibitors of histone deacetylases (HDAC), with consequences for chromosome remodeling, cell cycle arrest and selective toxicity in cultured melanoma cells compared with normal melanocytes. Few effects have been found on pigmentation, except paradoxically the down-regulation of TRP-1. Of the many genes regulated by HDAC inhibitors, induction of p21(WAF1/Cip1) is the most consistent finding and is associated with G(1) or G(2) phase blocks. Some melanoma cell lines appear to lack an HDAC inhibitor-specific G(2) checkpoint and viability is thus compromised by dividing with inappropriately-modified chromatin. Most cultured melanoma cells undergo apoptosis following treatment with HDAC inhibitors, via a mitochondrial and caspase-dependent pathway. However, the molecular mechanism may vary with cell line and HDAC inhibitor class. Tumor selectivity cannot yet be attributed to specific types or levels of HDACs, nor has the possibility of acetylation of non-histone targets been excluded. Elucidation of these complexities may be rewarding, in terms of directing the multiple consequences of inhibiting histone deacetylation towards overcoming the therapeutic problems of melanoma heterogeneity and emergence of resistance. Success in the clinic may require combination with agents that synergize with the cell cycle blocking and pro-apoptotic action of HDAC inhibitors. PMID- 15892713 TI - WNT1 and WNT3a promote expansion of melanocytes through distinct modes of action. AB - Summary WNT1 and WNT3a have been described as having redundant roles in promoting the development of neural crest-derived melanocytes (NC-Ms). We used cell lineage restricted retroviral infections to examine the effects of WNT signaling on defined cell types in neural crest cultures. RCAS retroviral infections were targeted to melanoblasts (NC-M precursor cells) derived from transgenic mice that express the virus receptor, TVA, under the control of a melanoblast promoter (DCT). As expected, over 90% of DCT-TVA+ cells expressed early melanoblast markers MITF and KIT. However, by following the fate of infected cells in standard culture conditions, we find that only 5% of descendents were NC-Ms. The majority of the descendents were not NC-Ms, but expressed smooth muscle cell markers, demonstrating that mammalian melanoblasts are not committed to the NC-M lineage. RCAS infection of DCT-TVA+ cells demonstrated that overexpression of canonical WNT signaling genes (betaCAT, WNT3a or WNT1) can increase NC-M numbers in an endothelin dependent manner. However, WNT1 and WNT3a have different modes of action with respect to melanoblast fate. Intrinsic over-expression of betaCAT or WNT3a can increase NC-M numbers by biasing the fate of DCT-TVA+ cells to NC Ms. In contrast, the DCT-TVA+ melanoblasts cannot respond to WNT1 signaling and do not alter their fate towards NC-M. Instead, WNT1 only increases NC-M numbers through paracrine signaling on melanoblast precursors to increase the numbers of neural crest cells that become NC-Ms. PMID- 15892714 TI - Ednrb2 orients cell migration towards the dorsolateral neural crest pathway and promotes melanocyte differentiation. AB - Endothelin receptors B (Ednrb) are involved in the development of the enteric and melanocytic lineages, which originate from neural crest cells (NCCs). In mice, trunk NCCs and their derivatives express only one Ednrb. In quail, trunk NCCs express two Ednrb: Ednrb and Ednrb2. Quail Ednrb is expressed in NCCs migrating along the ventral pathway, which gives rise to the peripheral nervous system, including enteric ganglia. Ednrb2 is upregulated in NCCs before these cells enter the dorsolateral pathway. The NCCs migrating along the dorsolateral pathway are melanocyte precursors. We analyzed the in vitro differentiation and in ovo migration of mouse embryonic stem (ES) cells expressing and not expressing Ednrb2. We generated a series of transfected ES cell lines expressing Ednrb2. This receptor, like Ednrb, oriented genuine ES cells towards melanocyte lineage differentiation in vitro. The in ovo migration of Ednrb2-expressing ES cells was massively oriented towards the dorsolateral pathway, unlike that of WT or Ednrb expressing ES cells. Thus, Ednrb2 is involved in melanoblast differentiation and migration. PMID- 15892715 TI - Establishment of a kit-negative cell line of melanocyte precursors from mouse neural crest cells. AB - We previously established a mouse neural crest cell line named NCCmelb4, which is positive for Kit and negative for tyrosinase. NCCmelb4 cells were useful to study the effects of extrinsic factors such as retinoic acids and vitamin D(3) on melanocyte differentiation, but in order to study the development of melanocytes from multipotent neural crest cells, cell lines of melanocyte progenitors in earlier developmental stages are needed. In the present study, we established an immortal cell line named NCCmelb4M5 that was derived from NCCmelb4 cells. NCCmelb4M5 cells do not express Kit and are immortal and stable in the absence of Kit ligand. They are positive for melanocyte markers such as tyrosinase-related protein 1 and DOPAchrome tautomerase and they contain stage I melanosomes. Interestingly, glial fibrillary acidic protein, which is a marker for glial cells, is also positive in NCCmelb4M5 cells, while NCCmelb4 cells are negative for this protein. Immunostaining and a cell ELISA assay revealed that 12-O tetradecanoylphorbol 13-acetate (TPA) and cholera toxin (CT) induce Kit expression in NCCmelb4M5 cells. Real-time polymerase chain reaction analysis also demonstrated the induction of Kit mRNA by TPA and CT. Microphthalmia-associated transcription factor mRNA is simultaneously enhanced by the same treatment. Kit induced by TPA/CT in NCCmelb4M5 cells disappeared after the cells were subcultured and incubated without TPA/CT. These findings show that NCCmelb4M5 cells have the potential to differentiate into Kit-positive melanocyte precursors and may be useful to study mechanisms of development and differentiation of melanocytes in mouse neural crest cells. PMID- 15892716 TI - Melanoma cell lines are susceptible to histone deacetylase inhibitor TSA provoked cell cycle arrest and apoptosis. AB - Melanoma is the most aggressive of skin cancers because of its high resistance to currently available therapy. Although melanoma cells often retain wild-type p53 tumour suppressor protein and express it at high levels, the p53 mediated apoptosis pathway is suppressed. Histone deacetylase (HDAC) inhibitors are a promising group of compounds inducing differentiation, growth arrest and apoptosis in tumour cells in preclinical studies. We have studied the cellular effects of trichostatin A (TSA), a HDAC inhibitor, in a panel of melanoma cell lines and its mechanism of action in relation to p53. TSA stabilized wild-type p53, but p53 protein accumulation was overridden by simultaneous downregulation of p53 mRNA leading to a decrease in p53 protein. While growth arrest was induced in all cell lines studied and apoptosis in most (6/7), these cellular effects were independent of the p53 status of the cells. Inhibiting p53 function by a dominant negative p53 (p53(175His)) confirmed that the HDAC inhibitor induced apoptosis was independent of wild-type p53, even though TSA slightly activated p53 in a reporter assay. The results indicate that while the action of TSA is independent of p53, the activation of the apoptosis pathway by the HDAC inhibitors may provide therapeutic approaches for melanoma treatment. PMID- 15892717 TI - Dynamic regulation of the human dopachrome tautomerase promoter by MITF, ER-alpha and chromatin remodelers during proliferation and senescence of human melanocytes. AB - Senescent cells are known to display altered gene expression of differentiation associated genes. We have previously demonstrated that the melanocyte transcriptional regulator microphthalmia-associated protein (MITF) is down regulated in senescent melanocytes. Since virtually nothing is known regarding the differentiated function of senescent melanocytes, we analyzed the transcriptional regulation of Dopachrome tautomerase (DCT), a member of the tyrosinase gene family, in proliferating and in senescent human melanocytes. Computational analysis of the region containing the M-box that includes the MITF CATGTG binding motif demonstrated that this sequence overlaps with the estrogen receptor alpha (ER-alpha), USF-1, TFE-3, Isl-1 and AP-1 binding elements. Electrophoresis gel-shift analysis using an oligonucleotide containing MITF and ERE elements identified MITF and ER-alpha complexes in proliferating melanocytes, whereas only ER-alpha complexes were detected in senescent cells. Importantly, a promoter-reporter analysis demonstrated that the coactivator p300/CBP switched MITF from a repressor to an activator of DCT transcription. p300/CBP was also required by ER-alpha and MITF to induce high, synergistic activation of the DCT promoter. We have also found that transcription of the DCT gene is differentially regulated by major melanocyte mitogens. In contrast to the activating effect of cAMP inducers, 12-O-tetradecanoylphorbolacetate (TPA) was a potent repressor of DCT transcription, suggesting that this gene can be differentially regulated by multiple environmental signals and promoter context. In support of this conclusion, trichostatin A, a histone deacetylase inhibitor, counteracted the TPA mediated repression, and restored high levels of DCT protein in cultured melanocytes. We conclude that senescent melanocytes display dramatic changes in the expression of differentiation-related proteins; such changes may in turn result in altered melanocyte function and survival to environmental stresses. PMID- 15892718 TI - An oxygen transporter hemocyanin can act on the late pathway of melanin synthesis. AB - 5,6-Dihydroxyindole (DHI) and 5,6-dihydroxyindole-2-carboxylic acid (DHICA) are precursors of eumelanin. The effects of crustacean hemolymph proteins on these eumelanin-related metabolites were investigated. Zymogram analysis indicated that polymers of hemocyanin (Hc) subunits converted DHI into black pigment while no effects were observed using DHICA as a substrate. Spectrum changes for mixtures of purified Hc and DHI showed a profile similar to oxidized DHI by mushroom tyrosinase while Hc had only slight effects on DHICA. Typical inhibitors of tyrosinase and phenoloxidase severely hampered the production of oxidized DHI. Taken together with previous results, these data indicate that Hc plays a crucial role in the conversion of DHI in the hemolymph of crustaceans, which promotes late reactions in the melanin synthetic pathway as well as early reactions (oxidation of tyrosine and DOPA to dopaquinone). PMID- 15892719 TI - Eumelanin and pheomelanin concentrations in human epidermis before and after UVB irradiation. AB - Pheomelanin is widely thought to be causally related to susceptibility to the harmful effects of ultraviolet radiation: epidemiological studies show that those with a higher ratio of pheomelanin to eumelanin in hair have higher rates of melanoma, and work in mouse and cell culture shows that pheomelanin generates excess free radicals after UVR exposure. By contrast, based on measurements of eumelanin and pheomelanin in human skin, before and following irradiation, we now report that both pheomelanin and eumelanin are positively related to skin colour, and by inference, inversely with cancer susceptibility. The ratio of melanin classes is similar in people with widely different cancer rates and UVR sensitivity. Although our numbers are small, our results extend previous work in man, and lead us to speculate that factors other than the amount of pheomelanin may be important in determining UVR susceptibility in persons with red hair. PMID- 15892721 TI - Nursing identity and difference. PMID- 15892722 TI - Modes of rationality in nursing documentation: biology, biography and the 'voice of nursing'. AB - This article is based on a discourse analysis of the complete nursing records of 45 patients, and concerns the modes of rationality that mediated text-based accounts relating to patient care that nurses recorded. The analysis draws on the work of the critical theorist, Jurgen Habermas, who conceptualised rationality in the context of modernity according to two types: purposive rationality based on an instrumental logic, and value rationality based on ethical considerations and moral reasoning. Our analysis revealed that purposive rationality dominated the content of nursing documentation, as evidenced by a particularly bio-centric and modernist construction of the workings of the body within the texts. There was little reference in the documentation to central themes of contemporary nursing discourses, such as notions of partnership, autonomy, and self-determination, which are associated with value rationality. Drawing on Habermas, we argue that this nursing documentation depicted the colonisation of the sociocultural lifeworld by the bio-technocratic system. Where nurses recorded disagreements that patients had with medical regimes, the central struggle inherent in the project of modernity became transparent--the tension between the rational and instrumental control of people through scientific regulation and the autonomy of the subject. The article concludes by problematising communicative action within the context of nursing practice. PMID- 15892723 TI - The deconstructing angel: nursing, reflection and evidence-based practice. AB - This paper explores Jacques Derrida's strategy of deconstruction as a way of understanding and critiquing nursing theory and practice. Deconstruction has its origins in philosophy, but I argue that it is useful and relevant as a way of challenging the dominant paradigm of any discipline, including nursing. Because deconstruction is notoriously difficult to define, I offer a number of examples of deconstruction in action. In particular, I focus on three critiques of reflective practice by the meta-narrative of evidence-based practice (EBP) and attempt to show how those critiques can be directed back at EBP itself. I conclude with the observation that EBP is open to many of the criticisms that it directs at other discourses, including problems of a lack of empirical evidence, of distortions due to memory, and of falsification of the 'facts'. PMID- 15892724 TI - Men, culture and hegemonic masculinity: understanding the experience of prostate cancer. AB - Following a diagnosis of, and treatment for prostate cancer, there is an expectation that men will cope with, adjust to and accept the psychosocial impact on their lives and relationships. Yet, there is a limited qualitative world literature investigating the psychosocial experience of prostate cancer, and almost no literature exploring how masculinity mediates in such an experience. This paper will suggest that the experience of prostate cancer, the process by which it is investigated, and the way in which it is understood has been shaped by an essentialist interpretation of gender, exemplified by hegemonic masculinity as the archetypal mechanism of male adaptation. In response to this static and limiting view of masculinity, this paper will offer a reframe of hegemonic masculinity. This reframe, being more aligned with common experience, will portray masculinity as a dynamic and contextual construct, better understood as one of a number of cultural reference points around which each man organises and adopts behaviour. It will be suggested that the extant literature, in being organised around hegemonic masculinity, obfuscates the experience of prostate cancer and acts to render covert any collateral masculinities, public or private, that may also be operating. PMID- 15892725 TI - The language of 'experience' in nursing research. AB - This paper is an analysis of how the signifier 'experience' is used in nursing research. We identify a set of issues we believe accompany the use of experience but are rarely addressed. These issues are embedded in a spectrum that includes ontological commitments, visions of the person/self and its relation to 'society', understandings of research methodology and the politics of nursing. We argue that a poststructuralist understanding of the language of experience in research opens up additional ways to analyze the relationship between the conduct of nursing research and cultural/political commitments. PMID- 15892726 TI - Phenomenology as rhetoric. AB - The literature on 'nursing phenomenology' is driven by a range of ontological and epistemological considerations, intended to distance it from conventionally scientific approaches. However, this paper examines a series of discrepancies between phenomenological rhetoric and phenomenological practice. The rhetoric celebrates perceptions and experience; but the concluding moment of a research report almost always makes implicit claims about reality. The rhetoric insists on uniquely personal meanings; but the practice offers blank, anonymous abstractions. The rhetoric invites us to believe that knowing is subjective and involved, but at the same time it recommends a technique (bracketing) which can only represent a crude, and entirely misconceived, gesture towards objectivity. Finally, the rhetoric claims that generalisation is beside the point; but the majority of researchers generalise anyway. In quietly ignoring their own rhetoric, 'phenomenologists' appropriate scientific prerogatives illegitimately. For their methods do not entitle them to lay claim to anything resembling 'objectivity', or generalisability, or 'reality', or theoretical abstraction. Like other researchers, they want to talk in generalisable terms about reality; they want to be objective, they want to do theory. But they are saddled with a philosophy that is disabling, because it says they can only talk about perceptions, and meanings, and uniqueness. PMID- 15892727 TI - Nursing practice as bricoleur activity: a concept explored. AB - The debates concerning the nature of nursing practice are often rooted in tensions between artistic, scientific and magical/mythical practice. It is within this context that the case is argued for considering that nursing practice involves bricoleur activity. This stance, which is derived from the work of Levi Strauss, conceives elements of nursing practice as an embodied, bricoleur practice where practitioners draw on the 'shards and fragments' of the situation at-hand to resolve the needs of the individual patient for whom they care. This conceptualisation of nursing practice will be analysed with a particular emphasis on its implication for nursing epistemology, pedagogy and praxis. The evidence to support this argument is drawn from empirical work that investigated nurses' use of intuition, the work of Levi-Strauss, and issues in nursing epistemology and ontology. The paper itself is written from the perspective of a bricoleur who uses 'bits and pieces' from the domains of nursing, philosophy, psychology, education, sociology and anthropology. PMID- 15892728 TI - Mediating the meaning of evidence through epistemological diversity. AB - Nursing's disciplinary recognition of 'multiple ways of knowing' reflects an epistemological diversity that supports nursing praxis. Nursing as praxis offers a conceptual way to explore what it is about the interface of practice, knowledge and evidence in nursing that distinguishes us as a discipline. I suggest that the relationship between evidence and knowledge is defined and mediated by the same epistemological diversity that supports nursing as praxis. Just as the meaning and truth-value of evidence is evaluated from within the body of existing disciplinary knowledge, new evidence may prompt an evaluation of the meaning and truth-value of extant nursing knowledge. Nursing practice that relies on scientific evidence as a singular basis of practice knowledge is susceptible to exploitation by the diverse agendas operating within an ideology of evidence based practice and the healthcare system. Mediating the meaning of evidence for nursing practice through acknowledgement of the diverse epistemologies that underpin nursing knowledge will contribute to a disciplinary-specific definition of what constitutes evidence for nursing, and will better direct how evidence is integrated into a disciplinary body of knowledge. PMID- 15892729 TI - In the public interest: autonomy and resistance to methods of standardising nurses' advice and practices from a health call centre in Perth, Western Australia. AB - The history of nursing is replete with examples of nurses battling for autonomy over their education, knowledge and work practices. The latest battleground is HealthDirect, Australia's first medial call centre, where nurses are required to meet externally imposed clinical standards while satisfying legal and financial obligations. These objectives are arguably achieved when nurses assess callers' health problems via computerised algorithms that determine an appropriate plan of action. That way, nurses' subjective responses to callers are ruled out. To ensure nurses comply with the standard processes, calls are monitored randomly and surreptitiously in formal and informal ways. This paper explores how nurses respond to standard procedures and surveillance, in order to argue that nurses' input partially drives reform processes. Nurses continue to seek autonomy over the advice they give and how their work is evaluated but are constrained by structural power relations. PMID- 15892730 TI - Taking account of local culture: limits to the development of a professional ethos. AB - The need to extend the discussion of culture in the study of nursing, combined with an enthusiasm for the possibility of viewing nursing from a new perspective, provides the impetus for this study. Based on fieldwork undertaken in the intensive care unit (RICU) of a major public hospital in Rome (Italy), this paper explores some of the key aspects of the social relations and local staff culture of one particular group of Italian nurses. In a climate of globalization, where the deployment of dominant Anglo-American ideas is difficult to counter, the RICU presents as a setting which challenges the widespread assumptions of universal standards of nursing practice. By building a picture of the working world of these particular nurses, we are assisted in our understanding of nursing practice as a local cultural activity. In exploring the significance of local culture this paper brings into question the universality of the current paradigm of professionalism and professional identity, and emphasizes the value of acknowledging local culture. PMID- 15892731 TI - A feminist utilitarian perspective on euthanasia: from Nancy Crick to Terri Schiavo. PMID- 15892732 TI - Carbon dioxide increases acid resistance in Escherichia coli. AB - AIMS: To investigate how carbon dioxide affects the acid resistance of Escherichia coli. METHODS AND RESULTS: Escherichia coli W3110 was grown in minimal EG medium at pH 7.5, and cells were adapted at pH 5.5 at 37 degrees C with and without supply of carbon dioxide and nitrogen gases. The number of colonies grown on LB medium was measured after cells were challenged in minimal EG medium of pH 2.5 at 37 degrees C under various conditions. When carbon dioxide was supplied at both the acid adaptation and challenge stages, 94% of cells survived after the acid challenge for 1 h, while the survival rates were 50 and 67% when nitrogen gas and glutamate were supplied respectively. After the acid challenge for 3 h, the survival rate observed with the carbon dioxide gas supply was again 2.5-fold higher than those with the nitrogen gas supply. CONCLUSION: Carbon dioxide was shown to participate in the maintenance of high viability under acidic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides useful information for research into bacterial pathogenesis, fermentation and food preservation. PMID- 15892733 TI - Optimization of glutamate concentration and pH for H production from volatile fatty acids by Rhodopseudomonas capsulata. AB - AIMS: This study attempted to employ response surface methodology (RSM) to evaluate the effects of glutamate concentration and pH on H(2) production from volatile fatty acids by Rhodopseudomonas capsulata. METHODS AND RESULTS: A mixture of acetate, propionate and butyrate was used as a carbon source for the H(2) production by R. capsulata. The H(2) yield and H(2) production rate were strongly affected by the glutamate concentration, pH and their interaction. The predicted maximum H(2) yield of 0.534 was obtained when glutamate concentration and pH were 6.56 mmol l(-1) and 7.29 respectively. On the contrary, the maximum H(2) production rate of 18.72 ml l(-1) h(-1) was achieved at a glutamate concentration of 7.01 mmol l(-1) and pH 7.31. CONCLUSIONS: Taking H(2) yield and H(2) production rate together into account, a glutamate concentration of 6.56 7.01 mmol l(-1) and pH of 7.29-7.31 should be selected for H(2) production from a mixture of acetate, propionate and butyrate by R. capsulata. SIGNIFICANCE AND IMPACT OF THE STUDY: The RSM was a useful tool for maximizing H(2) production by photosynthetic bacteria (PSB). PMID- 15892734 TI - Multiplex PCR assay for toxinotyping Clostridium perfringens isolates obtained from Finnish broiler chickens. AB - AIMS: The aim of the study was to determine the presence of genes coding for alpha (cpa), beta (cpb), epsilon (etx), iota (iA) and enterotoxin (cpe) from Clostridium perfringens broiler chicken isolates, using multiplex PCR assay established in the study. METHODS AND RESULTS: The multiplex PCR assay was shown to be specific when tested with 10 C. perfringens strains representing different toxin types, and 15 strains of other bacterial species. All 118 broiler chicken C. perfringens isolates were shown to carry the cpa gene but not cpb, etx, iap or cpe genes, signifying that all isolates represented type A and were cpe-negative. CONCLUSIONS: The assay established in the study enables the simultaneous detection of the major toxin genes and the cpe gene from C. perfringens isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study offers a new primer pair for detecting cpa, combined with a multiplex PCR assay. In addition, the study provides data of the presence of different toxin genes in C. perfringens isolates obtained from broiler chickens. PMID- 15892735 TI - Effect of oxidative stress and exogenous beta-carotene on sclerotial differentiation and carotenoid yield of Penicillium sp. PT95. AB - AIMS: To determine the effect of oxidative stress and exogenous beta-carotene on sclerotial differentiation and carotenoid yield of Penicillium sp. PT95. METHODS AND RESULTS: In this experiment, high oxidative stress was applied by inclusion of FeCl(3) (10 micromol l(-1)) in the growth medium and by light exposure. Low oxidative stress was applied by omitting iron from the growth medium and by incubation in the dark. Supplementation of exogenous beta-carotene (as antioxidant) to the basal medium caused a concentration-dependent delay of sclerotial differentiation (up to 72 h), decrease of sclerotial biomass (up to 43%) and reduction of carotenoid yield (up to 92%). On the contrary, the exogenous beta-carotene also caused a concentration-dependent decrease of lipid peroxidation in colonies of this fungus. CONCLUSIONS: Under high oxidative stress growth condition, the sclerotial biomass and carotenoid yield of PT95 strain in each plate culture reached 141 mg and 30.03 microg, which were 1.53 and 3.51 times higher respectively, than that at low oxidative stress growth condition. SIGNIFICANCE AND IMPACT OF THE STUDY: These data prompted us to consider that in order to attain higher sclerotial biomass and pigment yield, the strain PT95 should be grown under high oxidative stress and in the absence of antioxidants. PMID- 15892736 TI - Persistence of Campylobacter jejuni on surfaces in a processing environment and on cutting boards. AB - AIMS: The objectives of the study were to determine the spread and persistence of Campylobacter in a poultry processing plant and to provide a quantitative estimate of the survival of Campylobacter jejuni on the surface of a cutting board. METHODS AND RESULTS: Several contact surfaces in a poultry processing plant were sampled before the start of processing, after 30 min and after 120 min. Next, the survival of four C. jejuni strains was studied on a beech and polypropylene cutting board during 120 min. CONCLUSIONS: A rapid introduction and spread of Campylobacter in a well cleaned processing plant as well as a significant survival in time on the example of a cutting board is shown. SIGNIFICANCE AND IMPACT OF THE STUDY: The need to prevent cross-contamination in the food processing and preparation area and the importance of an integrated approach throughout the whole food chain to control transmission of Campylobacter is highlighted. PMID- 15892737 TI - Antisense inhibition of xylitol dehydrogenase gene, xdh1 from Trichoderma reesei. AB - AIMS: To inhibit xylitol dehydrogenase (XDH) in Trichoderma reesei by antisense inhibition strategy and construct novel strains capable of accumulating xylitol. METHODS AND RESULTS: The xdh1 antisense expression plasmid pGTA-xdh was constructed by inserting xdh1 DNA fragment inversely between the gpdA promoter and the trpC terminator from Aspergillus nidulans into a pUC19 plasmid backbone. Trichoderma reesei protoplasts were co-transformated with pGTA-xdh and hygromycin B resistance plasmid pAN7-1. Of 20 transformants screened from the selective medium, one transformant with the highest xylitol accumulation, designated ZY15, showed a distinct reduction (c. 52%) in XDH activity compared with the original strain Rut-C30. The results of Southern hybridization and PCR assay showed that the antisense expression cassette of xdh1 was integrated into the genome of T. reesei. The RT-PCR analysis proved that antisense RNA effectively inhibited XDH expression (c. 65%). Xylitol accumulation (2.37 mg ml(-1)) of ZY15 was five times higher than that (0.46 mg ml(-1)) of the original strain Rut-C30. CONCLUSIONS: Strain ZY15 successfully downregulated XDH production and exhibited xylitol accumulation in xylose liquid medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributed to the budding field of fungal genetics in two points. First, it confirmed that antisense RNA strategy could be used as a means of reducing gene expression in the filamentous fungus T. reesei. Secondly, it verified that the strategy appears most promising for creating novel filamentous fungi strains capable of accumulating intermediary metabolites. PMID- 15892738 TI - Durancin L28-1A, a new bacteriocin from Enterococcus durans L28-1, isolated from soil. AB - AIMS: To isolate, characterize and identify bacteriocins from lactic acid bacteria in soil. METHODS AND RESULTS: Thirty-four acid-producing bacteria were isolated from 87 soil samples. Antibacterial activities were detected, and one strain, L28-1 produced a bacteriocin that was active against some Gram-positive bacteria. L28-1 was identified as Enterococcus durans by 16S rDNA sequence analysis and API50CHL. This bacteriocin did not lose its activity after autoclaving (121 degrees C for 15 min), but was inactivated by protease K. The bacteriocin was purified by hydrophobic column chromatography, and Sep-Pak C(18). Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the partially purified bacteriocin contained numerous protein bands. Two bands that displayed antibacterial activities were c. 3.4 and 2.5 kDa in size. In this work, the 3.4-kDa bacteriocin was analysed with N-terminal amino acid and DNA sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis. The results indicated that the 3.4-kDa bacteriocin of Ent. durans L28-1 is a new natural enterocin variant. CONCLUSIONS: Enterococcus durans L28-1 produced a new bacteriocin. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports a novel bacteriocin that is produced by Ent. durans that has potential for use as a food preservative. PMID- 15892739 TI - Identification of sulI allele of dihydropteroate synthase by representational difference analysis in Haemophilus parasuis serovar 2. AB - AIMS: Identification of genes differentially present in Haemophilus parasuis serovar 2 by representational difference analysis (RDA). METHODS AND RESULTS: Bacterial genomic DNA was extracted, cleaved with Sau3AI and ligated to oligonucleotide adapter pair. The optimal tester (H. parasuis serovar 2)/driver ratio (H. parasuis serovars 1, 3 and 5) for the hybridization was established and the mixture was hybridized, and amplified by PCR. The products were cloned and transformed into Escherichia coli TOP10 cells and checked for specificity by Southern blotting analysis. The RDA subtractive technique yielded six bands ranging from 1500 to 200 bp, which were cloned into pCR II-TOPO vector and 40 clones were analysed. A fragment of 369 bp was specific for H. parasuis serovar 2, and showed 99% homology to sulI gene encoding for dihydropteroate synthase (dhps). The dhps gene conferring sulfonamide resistance was detected in H. parasuis serovar 2 but was absent in serovars 1, 3, 5 and in most of the Actinobacillus pleuropneumoniae serotypes (except serotype 7). CONCLUSION: sulI allele of dihydropteroate synthase has been identified in H. parasuis serovar 2 by RDA technique. SIGNIFICANCE AND IMPACT OF THE STUDY: The RDA technique seems to be an useful method for the identification of genes that are differentially present in H. parasuis, a respiratory pathogen of veterinary interest. PMID- 15892740 TI - Removal of micro-organisms in a small-scale hydroponics wastewater treatment system. AB - AIMS: To measure the microbial removal capacity of a small-scale hydroponics wastewater treatment plant. METHODS AND RESULTS: Paired samples were taken from untreated, partly-treated and treated wastewater and analysed for faecal microbial indicators, i.e. coliforms, Escherichia coli, enterococci, Clostridium perfringens spores and somatic coliphages, by culture based methods. Escherichia coli was never detected in effluent water after >5.8-log removal. Enterococci, coliforms, spores and coliphages were removed by 4.5, 4.1, 2.3 and 2.5 log respectively. Most of the removal (60-87%) took place in the latter part of the system because of settling, normal inactivation (retention time 12.7 d) and sand filtration. Time-dependent log-linear removal was shown for spores (k = -0.17 log d(-1), r(2) = 0.99). CONCLUSIONS: Hydroponics wastewater treatment removed micro organisms satisfactorily. SIGNIFICANCE AND IMPACT OF THE STUDY: Investigations on the microbial removal capacity of hydroponics have only been performed for bacterial indicators. In this study it has been shown that virus and (oo)cyst process indicators were removed and that hydroponics can be an alternative to conventional wastewater treatment. PMID- 15892741 TI - Decontamination with vaporized hydrogen peroxide is effective against Mycobacterium tuberculosis. AB - AIMS: To determine the efficacy of room fumigation with vaporized hydrogen peroxide (VHP) in decontamination of viable Mycobacterium tuberculosis. METHODS AND RESULTS: About 8 x 10(4)-2.3 x 10(6) CFU of M. tuberculosis H37Rv and M. tuberculosis Beijing were dried in 10-microl drops in tissue culture plates, placed in steam-permeable Tyvek pouches and distributed on laboratory surfaces. The room was exposed to VHP delivered by air conditioning. Different exposure conditions were tested. Exposure to VHP resulted in sterilization of the bacterial samples in three different test runs. CONCLUSIONS: VHP treatment is an effective means of reducing and eliminating room contaminations of M. tuberculosis. SIGNIFICANCE AND IMPACT OF THE STUDY: Fumigation with VHP represents an alternative to formaldehyde fumigation, particularly for decontamination of animal rooms in tuberculosis research laboratories. PMID- 15892742 TI - Characterization of cold-active pectate lyases from psychrophilic Mrakia frigida. AB - AIMS: This study was conducted to determine optimal conditions for pectate lyase (PL) production by two psychrophilic yeast strains and to compare the properties of the cold-active enzymes using mesophilic PL as reference enzyme. METHODS AND RESULTS: Two psychrophilic yeasts isolated from remote geographical locations (European Alps, north Siberia) produced extracellular cold-active PL. Both strains were identified as Mrakia frigida by analysis of ITS and large subunit (LSU) rRNA sequences. Maximum enzyme production occurred at a cultivation temperature of 1 or 5 degrees C. The apparent optimum for enzyme activity was observed at 30 degrees C and pH 8.5-9. The enzymes were thermolabile, but were resistant to repeated freezing and thawing. CONCLUSION: We describe for the first time alkaline PL-producing representatives of the yeast species M. frigida. The two strains produce cold-active PL with similar properties, but have a different enzyme production pattern. SIGNIFICANCE AND IMPACT OF THE STUDY: The enzymes described in this study could be useful for a wide range of applications, such as low-temperature pretreatment of wastewater containing pectic substances. PMID- 15892743 TI - The synergistic activity between Cry1Aa and Cry1c from Bacillus thuringiensis against Spodoptera exigua and Helicoverpa armigera. AB - AIMS: To investigate the interaction between two crystal proteins, Cry1Aa and Cry1C, for future development of biopesticides based on Bacillus thuringiensis, toxicities of the two individual proteins and in combinations have been determined against Spodoptera exigua and Helicoverpa armigera larvae, and synergism between the proteins has been evaluated using synergistic factor. METHODS AND RESULTS: SDS-PAGE showed that Cry1Aa and Cry1C proteins could be expressed in acrystalliferous B. thuringiensis 4Q7 strain, with molecular weights of 135 and 130 kDa respectively. The bioassay results indicated a synergistic activity between Cry1Aa and Cry1C against S. exigua and H. armigera, and the highest toxicities could be observed in the combination of Cry1Aa and Cry1C at a ratio of 1 : 1. CONCLUSION: The two toxins, Cry1Aa and Cry1C, interact synergistically to exhibit higher toxicity against S. exigua and H. armigera. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the investigation on the synergistic activity between two B. thuringiensis Cry1 toxins. It can be applied to the rational design of new generations of B. thuringiensis biopesticides and to strategies for management of resistant insects. PMID- 15892744 TI - The vinification of partially dried grapes: a comparative fermentation study of Saccharomyces cerevisiae strains under high sugar stress. AB - AIMS: The study of the fermentation performance of Saccharomyces cerevisiae strains under high sugar stress during the vinification of partially dried grapes. METHODS AND RESULTS: Microvinification of partially dried grape must with sugar concentration of 35 degrees Brix was performed using four commercial strains to carry out alcoholic fermentation. A traditional red vinification without nutrients addition was applied. Yeasts displayed different efficiency to convert sugar in ethanol and varied in glycerol yield. Sugar consumption and ethanol level were attested at 80-87% and 143.5-158.0 g l(-1) respectively. High correlation between sugar and assimilable nitrogen consumption rate was observed. Statistical treatment of data by principal component analysis highlighted the different behaviours that strains exhibited in regard to the production of higher alcohols and other compounds important to wine quality. CONCLUSIONS: Saccharomyces cerevisiae strains displayed appreciable capability to overcome osmotic stress and to yield ethanol fermenting high sugar concentration grape must in winemaking condition. SIGNIFICANCE AND IMPACT OF THE STUDY: The results provided insights on the strain contribution to wine quality subordinate to stress condition. This investigation is of applicative interest for winemaking and processing industry that use high sugar concentration musts. PMID- 15892745 TI - Regulation of pyrimidine synthesis in Pseudomonas resinovorans. AB - AIMS: To investigate the regulation of de novo pyrimidine biosynthesis in the bacterium Pseudomonas resinovorans ATCC 14235. METHODS AND RESULTS: The pyrimidine biosynthetic pathway enzymes were measured in cell extracts from P. resinovorans ATCC 14235 and from an auxotroph lacking orotate phosphoribosyltransferase activity. Pyrimidine biosynthetic pathway enzyme activities in ATCC 14235 were affected by the addition of pyrimidine bases to the culture medium. The de novo enzyme activities of the phosphoribosyltransferase mutant strain increased after pyrimidine starvation indicating possible repression of the pathway by a pyrimidine-related compound. Aspartate transcarbamoylase activity in ATCC 14235 was inhibited in vitro by ATP, UTP and pyrophosphate. CONCLUSIONS: Pyrimidine biosynthesis in P. resinovorans was regulated at the level of enzyme synthesis and at the level of activity for aspartate transcarbamoylase. Its regulation of enzyme synthesis seemed to be similar to what has been observed in the taxonomically related species Pseudomonas oleovorans. SIGNIFICANCE AND IMPACT OF THE STUDY: This study found that pyrimidine biosynthesis is regulated in P. resinovorans. This could prove helpful to future studies investigating polyhydroxyalkanoate production by the bacterium. PMID- 15892746 TI - Rapid detection of viruses using electrical biochips and anti-virion sera. AB - AIMS: Rapid detection and quantification of viruses is crucial in clinical practice, veterinary medicine, agriculture, basic research as well as in biotechnological factories. However, although various techniques were described and are currently used, development of more rapid, more sensitive and quantitative methods seems to be still important. METHODS AND RESULTS: Here we describe a method for rapid detection of viruses (using bacteriophages as model viruses), based on electrical biochip array technology with the use of antibodies against capsid proteins. CONCLUSIONS: Using the procedure developed in this work, we were able to detect 2 x 10(4) virions on the chip. The whole assay procedure takes c. 50 min and the assay is quantitative. SIGNIFICANCE AND IMPACT OF THE STUDY: This procedure may be useful in various approaches, including detection of bacteriophage contamination in bioreactors and possibly detection of toxin gene bearing phages or other viruses in food samples. PMID- 15892747 TI - Microbiological quality changes in the intestine of hybrid tilapia (Oreochromis niloticus x Oreochromis aureus) in fresh and frozen storage condition. AB - AIMS: The goal of this study was to monitor the quantitative and qualitative bacterial flora in the intestine of hybrid tilapia in fresh fish and fish kept in frozen storage conditions for 1 year. METHODS AND RESULTS: Quantitative and qualitative analyses of the bacterial flora associated with the intestine of hybrid tilapia (Oreochromis niloticus x Oreochromis aureus) in fresh fish and fish kept in frozen storage conditions for 1 year were carried out. In fresh and frozen fish, aerobic plate count (APC) ranged from 1.6 +/- 1.2 x 10(8) to 1.5 +/- 0.9 x 10(5) CFU g(-1) in the intestine of tilapia collected from pond 1, 8.7 +/- 2.3 x 10(7) to 6.5 +/- 3.8 x 10(4) CFU g(-1) in the intestine of tilapia from pond 2, and 1.9 +/- 2.9 x 10(8) to 6.2 +/- 2.8 x 10(4) CFU g(-1) in the intestine of tilapia from pond 3. APC for all the groups of fish decreased c. 2-log cycles after 1 months frozen storage; thereafter, counts slowly declined during frozen storage for 1 year. Altogether, 16 bacterial genera were identified: Gram negative rods (67%) dominated. Both in fresh and frozen conditions, four bacterial species viz. Shewanella putrefaciens, Corynebacterium urealyticum, Aeromonas hydrophila and Flavobacterium sp. were always present, with a prevalence of 10% in most cases. Shewanella putrefaciens was the most dominant organism (15% of the total isolates) throughout the studied period. During frozen storage some of the bacteria were not recovered, but most of the bacteria survived after prolonged freezing. CONCLUSIONS: This study describes the aerobic heterotrophic microflora found in the intestine of fresh and frozen tilapia. The unique aspect of this study concerns the data revealing the micro-organisms, which are viable after prolonged freezing. Contamination of edible portions of fish could originate from gastrointestinal sources. SIGNIFICANCE AND IMPACT OF THE STUDY: The present results may enhance knowledge in controlling the storage life of fish, and fish product quality. Bacterial activity is by far the most important factor influencing fish quality, so bacterial numbers can be used as an index of quality. Storage of frozen tilapia without evisceration could be avoided. PMID- 15892748 TI - Phenotypic and genotypic characterization of sorbitol-negative or slow-fermenting (suspected O157) Escherichia coli isolated from milk samples in Lombardy region. AB - AIMS: To investigate phenotypic and genotypic aspects of sorbitol-negative or slow-fermenting Escherichia coli, suspected to belong to O157 serogroup, isolated in Italy. METHODS AND RESULTS: Milk samples originating from goats and cows were screened for the presence of E. coli O157 with cultural methods. Sorbitol negative or slow-fermenting strains were subjected to phenotypic characterization, antibiotic resistance profiles, PCR reactions for detection of toxins (stx(1) and stx(2)) and intimin (eae(GEN) and eae(O157)) genes and clustering by pulsed field gel electrophoresis (PFGE). Only one strain revealed to be O157. Susceptibility to 11 antibiotics highlighted the high resistance to tetracycline (50%), sulfonamide and streptomycin (33%). The stx(2) gene was detected in two strains; only the strain identified as O157 exhibited an amplicon for both eae genes. PFGE identified seven distinct XbaI macrorestriction patterns at a similarity level of 41%. CONCLUSIONS: The use of sorbitol fermentation as cultural method is not sufficient for STEC discrimination while PCR assay proved to be a valuable method. SIGNIFICANCE AND IMPACT OF THE STUDY: The study reports presence of Shiga toxin-producing E. coli in raw milk, signalling a potential risk for humans. PMID- 15892749 TI - A new Hypocrea strain producing harzianum A cytotoxic to tumour cell lines. AB - AIMS: To identify a new fungal strain, Hypocrea sp. F000527 producing a trichothecene metabolite, harzianum A, and to evaluate its cytotoxicity to tumour cell lines. METHODS AND RESULTS: A fungal strain, F000527, with cytotoxic activity was identified as a new Hypocrea strain based on morphological characteristics and internal transcribed spacers rDNA sequence data. Harzianum A was isolated from wheat bran culture by 50% acetone extraction, silica gel chromatography, Sephadex LH-20 chromatography and HPLC. The chemical structures were determined by ESI- or HRFAB-MS and (1)H and (13)C-NMR analyses. Harzianum A showed cytotoxicity to HT1080 and HeLa cell lines with IC(50) value of 0.65 and 5.07 Lg ml(-1) respectively. CONCLUSIONS: Harzianum A with a chemical formula of C(23)H(28)O(6) was isolated from a new Hypocrea strain and showed moderate to strong cytotoxicity to human cancer cell lines. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of the production of cytotoxic harzianum A by a new Hypocrea strain. PMID- 15892750 TI - Effects of the gill monogenean Zeuxapta seriolae (Meserve, 1938) and treatment with hydrogen peroxide on pathophysiology of kingfish, Seriola lalandi Valenciennes, 1833. AB - Infections by the gill fluke Zeuxapta seriolae are a serious concern for sea cage aquaculture of kingfish, Seriola lalandi. The present study aimed to determine the pathophysiological effects of a progressive infection with Z. seriolae and the effects of treatment with hydrogen peroxide. For the progression of infection study, infected fish were taken from a sea cage farm, treated to remove parasites and then infected by cohabitation with heavily infected fish. Samples were taken at 2-week intervals for 8 weeks. Infection intensity peaked at 4 weeks post infection (mean intensity 565.9) and the number of mature worms (2 mm fixed length or larger) peaked at 6 weeks post-infection. Attachment of Z. seriolae appeared to cause little localized pathology; however, the occurrence of hyperplastic lamellae increased as the infection progressed. Haemoglobin concentrations were negatively correlated with Z. seriolae intensity and were lower than controls at 4 weeks (35.8% decrease) and 6 weeks (57.4% decrease) post infection. Blood lactate concentration and plasma osmolality increased throughout the course of infection. For the effect of treatment experiment, groups of infected and non-infected fish were sampled either before or after treatment with hydrogen peroxide. Treated fish from both infected and uninfected groups had increased plasma lactate, osmolality and pH compared with pre-treatment groups. Treatment with hydrogen peroxide appeared to have acute effects on fish health but the magnitude (e.g. lactate, osmolality) and extent of the effects (e.g. haemoglobin) was much less than that caused by chronic infection with Z. seriolae. PMID- 15892751 TI - Evaluation of an experimental Aeromonas salmonicida epidemic in chinook salmon, Oncorhynchus tshawytscha (Walbaum). AB - To determine the dynamics of the transmission of Aeromonas salmonicida ssp. salmonicida infection, chinook salmon, Oncorhynchus tshawytscha, were exposed to bacteria by cohabitation. The latent period (time between exposure and infectivity) was determined by exposing a group of chinook salmonid fingerlings to A. salmonicida by bath, then, at daily intervals, by holding five exposed (donor) fish with approximately 50 naive fish for 24 h. The latent period was 3 days post-infection and the time period between the initial exposure to bacteria and the beginning of bacterial shedding was 4.5 days for the same animals. The prevalence and intensity of infection in the donor fish, to which recipient fish were exposed, i.e. the level of exposure, was highly correlated with the development of disease in recipient (susceptible) chinook salmon (r2 = 0.57). An experiment was conducted to determine the daily progress of infection and development of a furunculosis epidemic among recipient fish by cohabiting a single exposed fish with 43 unexposed salmon. At daily intervals, all fish (in seven treatment tanks and one control tank daily) were sacrificed and tested for the presence of A. salmonicida in the kidney (n = 3520). Over 10 days, mean prevalence among recipient fish reached 75% and disease related mortality exceeded 50%. Bacterial concentrations in the water continued to increase over the duration of the experiment in concert with the number of infected animals present in the population. PMID- 15892752 TI - Influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum). AB - The influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied experimentally in the laboratory and at a fish farm. In experiment I, the effect of parasitic infection on columnaris disease was studied using F. columnare carrier fish. The fish were exposed to Diplostomum spathaceum cercariae and a set of other stressors in order to induce clinical columnaris infection. Parasitic infection and other stressors failed to induce the disease. Disease occurred when the fish were challenged with F. columnare, but D. spathaceum infection did not enhance the severity of the infection. In experiment II, the influence of rearing density and water temperature was studied. Overall mortality was highest in fish at normal rearing density with high temperature (+23 degrees C). At low temperature (+18 degrees C) mortality was not affected by rearing density, but the transmission of columnaris disease was faster at normal rearing density at both temperatures. This supports the view that reduction of fish density could be used in prevention of columnaris disease especially if water temperature is high. Because the lower rearing density can also decrease the transmission of ectoparasites and penetrating endoparasites, it could be an efficient tool in ecological disease management. PMID- 15892753 TI - A parasite survey of farmed Southern bluefin tuna, Thunnus maccoyii (Castelnau). AB - Farmed Southern bluefin tuna (SBT) were examined for parasites. Samples of harvest fish, mortalities and some fish showing clinical signs of disease were investigated. Targeted screening was conducted for a scuticociliate, Uronema nigricans, the myxosporean Kudoa sp. and a sanguinicolid digenean, Cardicola forsteri. General parasitological investigation revealed a diverse parasite community of didymozoid digeneans, two species of copepods, a polyopisthocotylean monogenean and larval cestodes. Targeted screening for U. nigricans exposed a low prevalence, most probably due to a lack of sensitivity in the test method. Few of the parasites examined pose a risk to the health of farmed SBT. PMID- 15892754 TI - Protection of crayfish, Cambarus clarkii, from white spot syndrome virus by polyclonal antibodies against a viral envelope fusion protein. AB - White spot syndrome virus (WSSV) is a large double-stranded DNA virus, causing considerable mortality in penaeid shrimp and other crustaceans. WSSV produces five major structural proteins, including two major envelope proteins, VP28 and VP19. To produce VP28 and VP19 as a single protein for antibody production, DNA sequences encoding both open reading frames were fused together and cloned into pET-22b(+) expression vector. The fusion protein, VP(19+28), was expressed in Escherichia coli, purified using Ni2+ His affinity chromatography and injected into a rabbit. Antiserum collected from the immunized rabbit was tested in vivo for ability to protect crayfish, Cambarus clarkii, from disease caused by WSSV. Fifteen days after challenge with WSSV, treatment with VP(19+28) antiserum gave 100% protection against disease in the ambient temperature range of 15-22 degrees C and 65% protection at a constant temperature of 26 degrees C. These results demonstrated VP(19+28) antiserum is effective in protection of crayfish from WSSV and confirmed that VP19 and VP28 play an important role in WSSV host infection. Targeting both VP19 and VP28 may be effective for the design of both immunotherapeutic medicines and reagents to detect WSSV. PMID- 15892755 TI - Changes in the innate immune response of Atlantic salmon, Salmo salar L., exposed to experimental infection with Neoparamoeba sp. AB - An experiment was conducted to determine the effect of Neoparamoeba sp. infection on the innate immune responses of Atlantic salmon. Atlantic salmon were experimentally infected with Neoparamoeba sp. and serially sampled 0, 1, 4, 6, 8 and 11 days post-exposure (dpe). Histological analysis of infected fish gill arches identified the presence of characteristic amoebic gill disease lesions as early as 1 dpe with a steady increase in the number of affected gill filaments over time. Immune parameters investigated were anterior kidney phagocyte function (respiratory burst, chemotaxis and phagocytosis) and total plasma protein and lysozyme. In comparison with non-exposed control fish basal respiratory burst responses were suppressed at 8 and 11 dpe, while phorbol myristate acetate stimulated activity was significantly suppressed at 11 dpe. Variable differences in phagocytic activity and phagocytic rate following infection were identified. There was an increase in the chemotactic response of anterior kidney macrophages isolated from exposed fish relative to control fish at 8 dpe. Total protein and lysozyme levels were not affected by Neoparamoeba sp. exposure. PMID- 15892756 TI - Fibrosarcoma in yellow perch, Perca flavescens (Mitchill). PMID- 15892757 TI - The morphology of Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) spores released from Fredericella sultana (Bryozoa: Phylactolaemata). PMID- 15892758 TI - Ultrastructural features of excretory calcinosis in the lobster, Homarus americanus Milne-Edwards. PMID- 15892759 TI - Intravenous catheter-associated Staphylococcus aureus bacteraemia: a common problem that can be prevented. PMID- 15892760 TI - Cannula-associated Staphylococcus aureus bacteraemia: outcome in relation to treatment. AB - BACKGROUND: Despite the frequency of cannula-associated Staphylococcus aureus bacteraemia (CASAB) there is uncertainty regarding the duration of treatment required. AIM: To determine the relationship between the duration and type of treatment for CASAB and subsequent relapse with deep-seated S. aureus infection. METHODS: We prospectively studied 276 patients with CASAB. Patients were followed for at least 8 weeks after completion of antibiotic treatment. Initial and subsequent isolates of S. aureus were compared using molecular methods to determine strain similarity. RESULTS: Initial mortality was 9% (26 of 276) and a complicating focus of infection presented during initial treatment in 6% (15 of 250) of the survivors. There were nine relapses of deep-seated infection from the strain causing the original infection. Relapses were equally common following peripheral CASAB and central CASAB. There was no relationship between the duration of treatment and the rate of relapse of deep-seated infection (P = 0.24). This observation held true regardless of whether the duration of treatment was analysed as < or = 7 versus > or = 8, < or =10 versus > or =11, or < or=14 versus > or =15 days (P = 0.62, 0.87 and 0.16, respectively). CONCLUSION: Episodes of peripheral CASAB pose an equal risk of relapse to central cannula related episodes. Although further studies are needed to determine the optimal treatment of CASAB, our study strongly suggests that more than 14 days treatment is excessive for most patients who respond promptly to cannula removal and antibiotic treatment. PMID- 15892761 TI - Elevated circulating levels of matrix metalloproteinase-9 and -2 in patients with symptomatic coronary artery disease. AB - BACKGROUND: Matrix metalloproteinases (MMP-9 and MMP-2) have been implicated in development of atherosclerosis and plaque rupture in acute coronary syndromes (ACS). AIM: To determine the relationship between circulating MMPs and symptomatic coronary artery disease. METHODS: Plasma levels of MMP-9 and MMP-2 were measured in patients with ACS, stable angina (SA) and in controls, using a quantitative gelatin zymography. These measurements were correlated with markers of systemic inflammation (hs-CRP) in all subjects and myocardial injury (troponin T) in patients with ACS. RESULTS: Plasma MMP-9 in ACS was greater than in SA, and was greater in SA than in controls (P < 0.01 ACS vs SA and controls, P < 0.01 SA vs control). Plasma MMP-2 was significantly greater in ACS than SA or controls (P < 0.01 vs SA and controls). There was strong overall relationship between hs-CRP and MMP-9 (r = 0.65, P < 0.0001) driven by a significant relationship in ACS patients (r = 0.58, P = 0.02), as there was no significant association in SA or controls. A weaker overall correlation was found between hs-CRP and MMP-2 (r = 0.39, P = 0.02), but no significant relationship was present for either of the two patient subgroups or controls. There was no correlation between levels of troponin T and MMP-9, MMP-2 or hs-CRP in ACS patients. CONCLUSION: Quantitative gelatin zymography identifies increased circulating levels of MMP-9 and MMP-2 in patients with symptomatic coronary disease. MMP-9 and MMP-2 are higher in ACS than SA patients and might have use as markers of plaque rupture or instability. The strong relationship between MMP-9 and hs-CRP in ACS patients suggests MMP-9 might be an additional marker and/or consequence of the inflammatory component in ACS. PMID- 15892762 TI - Cisplatin and gemcitabine induction chemotherapy followed by concurrent chemoradiotherapy or surgery for locally advanced non-small cell lung cancer. AB - BACKGROUND: We report a study of induction chemotherapy followed by concurrent chemoradiotherapy for stage IIIA/IIIB non-small cell lung cancer. METHODS: Patients received two cycles of induction chemotherapy with cisplatin 100 mg/m(2) on day 1 and gemcitabine 1000 mg/m(2) on days 1, 8, and 15 of a 28-day cycle. If the disease was resectable [corrected] surgery was followed with two further cycles. If unresectable, patients received cisplatin 100 mg/m(2) day 1, 29 with 5 fluorouracil 1000 mg/m(2) per 24 h continuous infusion for 96 h on days 2-5 and days 30-33 of the radiotherapy administration. Radiation therapy consisted of 63 Gy, 35 fractions, 7 weeks. RESULTS: Of 48 patients, 40% had a partial response to induction chemotherapy. Four of eleven patients with stage IIIA tumours had resectable disease. The remaining seven patients plus 37 with stage IIIB disease had chemoradiotherapy. Response at the completion of all therapy was 62% (IIIA 73%, IIIB 59%). For all patients the median survival was 15.3 months: 1 year and 3 years, 58% and 25%, respectively. Those with IIIB disease responding to induction chemotherapy had significantly superior survival to those that did not respond (37 months vs 11 months; P = 0.005). This remained significant from a landmark at 8 weeks after the start of treatment (P = 0.01). CONCLUSION: These results are equivalent to other studies using induction chemotherapy prior to concurrent chemoradiotherapy. Response to induction chemotherapy may have major prognostic significance. PMID- 15892763 TI - Occupational bladder cancer in New Zealand: a 1-year review of cases notified to the New Zealand Cancer Registry. AB - AIM: To identify which cases of adult bladder cancer notified to the New Zealand Cancer Registry in 2001 had a probable occupational cause. METHODS: Occupational Safety and Health (OSH), in conjunction with the Massey University Centre for Public Health Research, interviewed and obtained an occupational history for 210 (162 men, 48 women) cases. RESULTS: Of the 162 male cases (response rate 65%), 45 (28%) were considered to be 'probable' occupational cancers. Of the 48 female cases (response rate 76%), three cases (6%) were considered to be 'probable' occupational cancers. The largest occupational group for men was truck drivers, which made up 51% of probable cases. Other common groups were engineering and metal workers (18%), crop farmers/orchardists (7%), textile and leather workers (7%), painters/furniture finishers (7%), and plastics manufacturing workers (4%). The three female cases considered to be of occupational origin included two textile workers and one telephonist. CONCLUSIONS: The percentage of cases considered to be of occupational origin is similar to that reported in Europe and the United States, indicating that occupational cancer is a major occupational health problem in New Zealand as it is in other parts of the world. PMID- 15892764 TI - Update on the use of stem cells for cardiac disease. AB - Major advances have recently been made in our understanding of stem cell biology, and in the application of stem cells to treat cardiac disease. Resident cardiac stem cells have now been described and the long-accepted paradigm of the adult mammalian heart as an organ without regenerative capacity has been questioned. Various stem-cell-based approaches for ameliorating cardiac disease have been shown to be beneficial in animal models and are now being trialled in humans, with several phase I clinical studies already completed. Although these clinical studies lacked adequate placebo controls, they have consistently shown promising results. If confirmed by larger phase II/III trials, it is possible that within a few years a powerful new therapeutic option may be available for the burgeoning number of patients suffering from myocardial ischaemia and/or other cardiac disorders. PMID- 15892765 TI - Quality of care for women presenting with benign breast conditions. AB - The aim of the study was to investigate the management of women with benign breast problems. A consecutive sample of women (n = 194) was assessed who presented to public or private sector providers. The main reasons for referral were breast lumps (62%); 56% of women who attended the public sector did not receive any recommendation compared to 40% who attended the private sector and clinical/general practitioner reviews were recommended to more women in the private sector (54%). Reasons for the discrepancy between public and private patients require further investigation. PMID- 15892766 TI - Substance abuse in young patients with type 1 diabetes: easily neglected in complex medical management. AB - The use of recreational drugs has become increasingly popular among young people. As a centre caring for a large group of young patients with type 1 diabetes, we have become concerned about the number of patients presenting with drug-related metabolic problems. We present a case series highlighting the issues of substance abuse in young patients with type 1 diabetes. PMID- 15892767 TI - Rocks in the head: extensive intracranial calcification. PMID- 15892768 TI - Analysis and overview of the guidelines for assessing fitness to drive for commercial and private vehicle drivers. AB - Guidelines on fitness to drive were released by AUSTROADS and the National Road Transport Commission in September 2003. No recognised legal medical authority was cited. There are three parts in the document: (i) background information, (ii) specific medical conditions and (iii) appendices of relevant documents and contacts. This paper analyses the relevance of the guidelines for physicians and notes that the disclaimer exonerates its authors from potential repercussions. Guidelines for both private and commercial drivers are combined in the document and the basis for such delineation is defined. A lack of universal Australian standards with no State indicating the driver's responsibility to report changes in health standards on the issued licences is confirmed by the guidelines. Not all States indemnify physicians for reporting contrary to patients' wishes, while South Australia and the Northern Territory mandate reporting those at risk. Much of the language is patronizing, expecting '... conciliatory and supportive ...' behaviour even with recalcitrant patients. No allowance is made for patients who may not fulfil the guidelines but whom the doctor may consider fit to drive. Ambiguity regarding responsibility to report, as identified in the background section, may leave the doctor vulnerable for not reporting a patient who subsequently may cause injury. Attempt is made to differentiate the role of the specialist from the family general practitioner (GP), advocating specialists for commercial drivers, although this is largely left to the discretion of the GP. There is an implied onus on doctors to report all patients with the conditions under review. Some diagnoses, such as syncope, are discussed in different sections with application of conflicting limitations. Inappropriate language, such as reference to a seizure being '... an isolated non-epileptiform event ...', or withdrawal of medications failing to be restricted to anti-epileptic medications confounds interpretation. Some sections, such as that on sleep and epilepsy, are effectively analysed, while illnesses such as dementia are considered superficially. The guidelines are an attempt to assess fitness to drive, but contain serious flaws and provide limited information upon which to base decisions. Ambiguous language complicates application of the guidelines and places the health care professional at risk, despite a disclaimer protecting its authors. PMID- 15892769 TI - Docetaxel-induced myositis: report of a novel side-effect. PMID- 15892770 TI - Simultaneous presentation of rofecoxib-induced acute hepatitis and acute interstitial nephritis. PMID- 15892771 TI - Evolutionary morality and public health. PMID- 15892772 TI - The heart, beating or not! PMID- 15892774 TI - Twenty-four hour continuous infusion of amphotericin B for the treatment of suspected or proven fungal infection in haematology patients. PMID- 15892775 TI - Coeliac disease and pancreatitis. PMID- 15892777 TI - Children's perceptions of discrimination: a developmental model. AB - Discrimination affects millions of children in the United States and throughout the world. Although the topic is important for both theoretical and applied reasons, little developmental work has examined children's perceptions of discrimination directed toward themselves and others. A review of past theoretical and empirical work on the perception of discrimination is provided. Next, a developmental model of the perception of discrimination is offered. The model identifies developmental and individual differences expected to influence judgments about discrimination, as well as situational variables that are likely to support attributions to discrimination. PMID- 15892778 TI - Cortisol reactivity is positively related to executive function in preschool children attending head start. AB - This study examined relations among cortisol reactivity and measures of cognitive function and social behavior in 4- to 5-year-old children (N = 169) attending Head Start. Saliva samples for the assay of cortisol were collected at the beginning, middle, and end of an approximately 45-min testing session. Moderate increase in cortisol followed by down-regulation of this increase was positively associated with measures of executive function, self-regulation, and letter knowledge but not with measures of receptive vocabulary, emotion knowledge, or false belief understanding. Regression analysis indicates that executive function accounted for the association between cortisol reactivity and self-regulation and letter knowledge. PMID- 15892779 TI - Modifications in children's goals when encountering obstacles to conflict resolution. AB - Previous studies have demonstrated that children's goals are associated with their success in peer relationships. The current study extends earlier findings by examining changes in children's goals during hypothetical conflicts. Participants were 252 children ages 9 to 12 years old (133 boys, 119 girls). As predicted, children's goals changed significantly when they encountered obstacles to conflict resolution, and these changes were predictive of their subsequent strategy choices. Both aggressive- and submissive-rejected children were more likely to evidence antisocial changes in their goals, including an increased desire to retaliate. They also showed reluctance to forego instrumental objectives. Other findings highlighted the need to investigate the combinations of goals children pursue as predictors of their strategies and the quality of their peer relationships. PMID- 15892780 TI - When induction meets memory: evidence for gradual transition from similarity based to category-based induction. AB - The ability to perform induction appears early; however, underlying mechanisms remain unclear. Some argue that early induction is category based, whereas others suggest that early induction is similarity based. Category- and similarity-based induction should result in different memory traces and thus in different memory accuracy. Performing induction resulted in low memory accuracy in adults and 11 year-olds, whereas 5-, and 7-year-olds were highly accurate (Experiment 1). After training to perform category-based induction, 5- and 7-year-olds exhibited patterns of accuracy similar to those of adults (Experiment 2). Furthermore, 7 year-olds, but not 5-year-olds, retained this training over time (Experiment 3). With novel categories, even adults performed similarity-based induction, exhibiting high memory accuracy (Experiment 4). These results suggest a gradual transition from similarity- to category-based induction with familiar categories. PMID- 15892781 TI - Infants' use of synchronized visual information to separate streams of speech. AB - In 4 studies, 7.5-month-olds used synchronized visual-auditory correlations to separate a target speech stream when a distractor passage was presented at equal loudness. Infants succeeded in a segmentation task (using the head-turn preference procedure with video familiarization) when a video of the talker's face was synchronized with the target passage (Experiment 1, N = 30). Infants did not succeed in this task when an unsynchronized (Experiment 2, N = 30) or static (Experiment 3, N = 30) face was presented during familiarization. Infants also succeeded when viewing a synchronized oscilloscope pattern (Experiment 4, N = 26), suggesting that their ability to use visual information is related to domain general sensitivities to any synchronized auditory-visual correspondence. PMID- 15892782 TI - What does it look like and what can it do? Category structure influences how infants categorize. AB - Despite a large body of research demonstrating the kinds of categories to which infants respond, few studies have directly assessed how infants' categorization unfolds over time. Four experiments used a visual familiarization task to evaluate 10-month-old infants' (N = 98) learning of exemplars characterized by commonalities in appearance or function. When learning exemplars with a common function, infants initially responded to the common feature, apparently forming a category, and only learned the individual features with more extensive familiarization. When learning exemplars with a common appearance, infants initially learned the individual features and apparently only formed a category with more extensive familiarization. The results are discussed in terms of models of category learning. PMID- 15892783 TI - Genetic influences in different aspects of language development: the etiology of language skills in 4.5-year-old twins. AB - The genetic and environmental etiologies of diverse aspects of language ability and disability, including articulation, phonology, grammar, vocabulary, and verbal memory, were investigated in a U.K. sample of 787 pairs of 4.5-year-old same-sex and opposite-sex twins. Moderate genetic influence was found for all aspects of language in the normal range. A similar pattern was found at the low end of the distribution with the exception of two receptive measures. Environmental influence was mainly due to nonshared factors, unique to the individual, with little influence from shared environment for most measures. Genetic and environmental influences on language ability and disability are quantitatively and qualitatively similar for males and females. PMID- 15892784 TI - Group norms, threat, and children's racial prejudice. AB - To assess predictions from social identity development theory (SIDT; Nesdale, 2004) concerning children's ethnic/racial prejudice, 197 Anglo-Australian children ages 7 or 9 years participated in a minimal group study as a member of a team that had a norm of inclusion or exclusion. The team was threatened or not threatened by an out-group that was of the same or different race. Consistent with SIDT, prejudice was greater when the in-group had a norm of exclusion and there was threat from the out-group. Norms and threat also interacted with participant age to influence ethnic attitudes, although prejudice was greatest when the in-group had an exclusion norm and there was out-group threat. The implications of the findings for SIDT are discussed. PMID- 15892785 TI - Cultural patterns in attending to two events at once. AB - This study examined cultural differences in children's simultaneous attention to 2 events versus quick alternation in which attending to 1 event momentarily interrupted attending to another. Thirty-one 6- to 10-year-old U.S. children of Mexican and European American heritage folded paper figures with 2 other first- to third-grade children and an adult. Mexican heritage children whose mothers averaged 7 grades of school more commonly attended to events simultaneously. European heritage and Mexican heritage children whose mothers had more than 12 grades of school more commonly alternated attention. Differences are interpreted in light of traditional indigenous North and Central American emphasis on learning through observation of ongoing events as well as school practices that emphasize learning by attending to one event at a time. PMID- 15892786 TI - Representations of attachment relationships in children of incarcerated mothers. AB - Representations of attachment relationships were assessed in 54 children ages 2.5 to 7.5 years whose mothers were currently incarcerated. Consistent with their high-risk status, most (63%) children were classified as having insecure relationships with mothers and caregivers. Secure relationships were more likely when children lived in a stable caregiving situation, when children reacted to separation from the mother with sadness rather than anger, and when children were older. Common reactions to initial separation included sadness, worry, confusion, anger, loneliness, sleep problems, and developmental regressions. Results highlight need for support in families affected by maternal imprisonment, especially efforts to promote stable, continuous placements for children, in addition to underscoring the importance of longitudinal research with this growing but understudied group. PMID- 15892787 TI - The development of nonverbal working memory and executive control processes in adolescents. AB - The prefrontal cortex modulates executive control processes and structurally matures throughout adolescence. Consistent with these events, prefrontal functions that demand high levels of executive control may mature later than those that require working memory but decreased control. To test this hypothesis, adolescents (9 to 20 years old) completed nonverbal working memory tasks with varying levels of executive demands. Findings suggest that recall-guided action for single units of spatial information develops until 11 to 12 years. The ability to maintain and manipulate multiple spatial units develops until 13 to 15 years. Strategic self-organization develops until ages 16 to 17 years. Recognition memory did not appear to develop over this age range. Implications for prefrontal cortex organization by level of processing are discussed. PMID- 15892788 TI - When you shouldn't do what you want to do: young children's understanding of desires, rules, and emotions. AB - This research investigated 4- through 7-year-olds' and adults' (n = 64) concepts about the emotional consequences of desire fulfillment versus desire inhibition in situations where people's desires conflict with prohibitive rules. Results revealed developmental increases in attributing positive or mixed emotions to story characters that make willpower decisions and negative or mixed emotions to characters that transgress. These developmental changes in emotion predictions were accompanied by age-related differences in emotion explanations. Whereas 4- and 5-year-olds largely explained emotions in relation to the characters' goals, 7-year-olds and adults further explained how rules and future consequences influence emotions. Results are discussed in relation to connections among children's psychological, deontic, and future-oriented reasoning about emotions as well as the development of self-control. PMID- 15892789 TI - Ego development and ethnic identity formation in rural American Indian adolescents. AB - Ethnic identity development was assessed in the context of ego development in 12- to 15-year-old students from a Southeastern American Indian community. Self protective was the modal level and was characterized by awareness of ethnic group membership but little exploration or self-reflection. Impulsive adolescents had the least developed ethnic identities and highest levels of interpersonal vulnerability. Conformist adolescents expressed positive feelings about ethnic group affiliation, described relationships as harmonious, but demonstrated moderate social anxiety. Postconformist adolescents had the highest levels of agency, social competence, and identity achievement, but also had high levels of psychological distress and family conflict. Adolescent identity strivings may be understood in context with the level and timing of psychosocial maturity, for which ego development appears a useful marker. PMID- 15892790 TI - The two faces of adolescents' success with peers: adolescent popularity, social adaptation, and deviant behavior. AB - This study assessed the hypothesis that popularity in adolescence takes on a twofold role, marking high levels of concurrent adaptation but predicting increases over time in both positive and negative behaviors sanctioned by peer norms. Multimethod, longitudinal data, on a diverse community sample of 185 adolescents (13 to 14 years), addressed these hypotheses. As hypothesized, popular adolescents displayed higher concurrent levels of ego development, secure attachment, and more adaptive interactions with mothers and best friends. Longitudinal analyses supported a popularity-socialization hypothesis, however, in which popular adolescents were more likely to increase behaviors that receive approval in the peer group (e.g., minor levels of drug use and delinquency) and decrease behaviors unlikely to be well received by peers (e.g., hostile behavior with peers). PMID- 15892792 TI - British Urological Foundation preceptorships to the Cleveland Clinic. PMID- 15892793 TI - The ascending (acquired undescended) testis: a phenomenon? PMID- 15892794 TI - Prostate-specific antigen testing in the new millennium. PMID- 15892795 TI - Is nutritional support beneficial to patients after cystectomy? PMID- 15892796 TI - What's new in the treatment of metastatic kidney cancer? PMID- 15892797 TI - Ejaculatory disorders and sexual function. PMID- 15892798 TI - The molecular basis of cryosurgery. PMID- 15892799 TI - Genitourinary medicine and surgery in prisons during the period of reform. PMID- 15892800 TI - The changing presentation of germ cell tumours of the testis between 1983 and 2002. AB - OBJECTIVE: To prospectively investigate the presentation of germ cell tumours (GCTs) of the testis in terms of stage or histology, as the incidence of this disease in increasing. PATIENTS AND METHODS: Patients diagnosed with GCT of the testis between 1983 and 2002 were categorised into three periods depending on the date of diagnosis of the GCT, and the presentational characteristics assessed. RESULTS There was a significant increase in the proportion of patients presenting with stage I disease (59% to 78%) and seminoma (43% to 58%) over this period. There was also a significant reduction in the size of the primary tumour (5 to 4 cm). CONCLUSION: A greater proportion of patients with GCT are presenting with stage I seminoma, the reasons for which are unclear, although earlier diagnosis through improved awareness of GCT may be important. PMID- 15892801 TI - Tumours of the ureter and renal pelvis treated with resection and renal autotransplantation: a study with up to 20 years of follow-up. AB - OBJECTIVE: To report long-term follow-up data from patients treated with resection of urothelial neoplasms of the upper urinary tract combined with autotransplantation of the kidney. PATIENTS AND METHODS: In a clinical and histopathological review of 23 patients who had 25 autotransplantations, they were followed for 7-20 years or until death. Nine patients had either a solitary kidney or bilateral renal pelvic tumours (group A) and 14 had a normal contralateral kidney (group B). RESULTS: Seven operations were unsuccessful, ending in nephrectomy. Of the nine patients in group A two with high-grade renal pelvic tumours survived with no dialysis and recurrences for 127 and 238 months, respectively. Three patients required haemodialysis 0-3 times weekly for 27, 85 and 108 months, respectively. Three patients with low-grade disease developed invasive recurrences in the autotransplanted kidney after 16, 27 and 90 months, respectively, and later died from the disease. One patient died in an accident after 14 months. Of the 14 patients in group B, one developed a deeply invasive recurrence in the autotransplanted kidney after 86 months, despite frequent controls. CONCLUSIONS: In patients with a normal contralateral kidney resection and renal autotransplantation is not indicated and might even be harmful, compared to standard nephroureterectomy. The operation might be beneficial in patients with solitary kidneys but other treatments should first be considered, including open or endoscopic resection, and nephroureterectomy and haemodialysis. PMID- 15892802 TI - A phase-1 study of sequential mitomycin C and 5-aminolaevulinic acid-mediated photodynamic therapy in recurrent superficial bladder carcinoma. AB - OBJECTIVE: To report a phase-1 study of patients with recurrent superficial bladder cancer treated with photodynamic therapy (PDT) using sequential mitomycin C and 5-aminolaevulinic acid (ALA). PATIENTS AND METHODS: Twenty-four patients were treated, the primary endpoint being the safety and tolerability of combined therapy at increasing doses of ALA and light. RESULTS: Mitomycin C instillation was followed by ALA concentrations of 6%, 8% or 10%; there was no effect on toxicity. The light dose, at a wavelength of 635 nm, was increased from zero to 25 J/cm(2), with the upper fluences producing transient symptoms. There were no episodes of skin photosensitivity or systemic toxicity. A total fluence of 25 J/cm(2) represented the upper light dose for the tolerability of this procedure by patients. There were no persistently high urinary symptom scores or reduction in functional bladder capacity up to > or =24 months of follow-up. In this group, cumulative tumour recurrences were none at 4, two at 8, six at 12, nine at 18 and 11 at 24 months after PDT, respectively. CONCLUSION: Sequential mitomycin C and ALA-PDT is a safe and well tolerated treatment, with potential for managing difficult-to-control superficial transitional cell carcinoma and carcinoma in situ of the bladder. PMID- 15892803 TI - Radical cystectomy in patients aged > or = 75 years: an updated review of patients treated with curative and palliative intent. AB - OBJECTIVE: To evaluate the morbidity and mortality of radical cystectomy in a group of unselected patients aged > or = 75 years who were treated with curative and palliative intent. PATIENTS AND METHODS: We retrospectively analysed 53 patients aged 75-90 years (median 78.8 years) who had radical cystectomies between May 1994 and July 2002. The patients were divided into two groups: 46 were treated with curative intent (group A) and seven with palliative intent (group B). The indications for cystectomy in group A were recurrent and otherwise therapy-resistant bladder cancer, severe irritative voiding symptoms, and recurrent macrohaematuria. The indications in group B were advanced pelvic malignancy with severe irritative voiding symptoms, severe pain, and recurrent macrohaematuria requiring blood transfusions. Patients were categorized according to the American Society of Anesthesiologists classification, with a score of II in 28 patients, III in 21 and IV in four. Complications and mortality before, during and after surgery, and the duration of hospital stay and clinical outcome, were assessed. RESULTS; The early mortality rate in group A was 4% (2/46); in group B two patients died after prolonged complications. The median (range) hospital stay was 28 (6-56) days, and was significantly longer in patients with complications, at a median (range) of 36 (6-70) days. The complication rates early and late after surgery in group A were 22% and 11%, respectively, and in group B, five of seven (early). The total median survival was 2 (0.33-7) years. CONCLUSIONS: Elderly people undergoing radical cystectomy have a greater risk of perioperative morbidity and mortality, especially those with very advanced pelvic malignancies who have had cystectomy with palliative intent. The incidence of early and late complications in patients treated with curative intent is acceptable, but the hospital stay is prolonged. PMID- 15892804 TI - Is photodynamic diagnosis using hypericin better than white-light cystoscopy for detecting superficial bladder carcinoma? AB - OBJECTIVE: To review the initial clinical results of photodynamic diagnosis (PDD) using hypericin (a new photosensitizer for PDD that helps to label flat urothelial tumours to facilitate biopsy) for the early detection of superficial bladder carcinoma, as flat noninvasive tumours of the bladder may be missed during conventional white-light cystoscopy (WLC) if there is bladder overdistension or ongoing cystitis. PATIENT AND METHODS: Between 1 January 2001 and 30 October 2004, 41 consecutive patients (mean age 66.1 years, sd 9.1, range 46-81) had transurethral resection for bladder cancer. Hypericin was introduced intravesically for 2 h before cystoscopy. Immediately after WLC, fluorescence cystoscopy (FC) was used at the same location and the same bladder site inspected using violet light. FC findings, e.g. positive or negative red fluorescence, were documented for each specific bladder site examined, and the exact location sampled for biopsy. RESULTS: The mean (sd, range) bladder capacity of the patients was 431 (86, 300-650) mL. In all, 179 biopsies were taken from the 41 patients; urothelial cancers were found in 41% (74) and 80% (33) had macroscopically visible bladder tumours; 40% (71) of the biopsies were positive under FC and 86% (61) of the 71 FC-positive biopsies showed cancer on histology. Twenty-five biopsies (14%) were positive on FC but not WLC. PDD testing with hypericin had a sensitivity of 82% (61/74) and specificity of 91% (95/105), vs WLC, at 62% (46/74) and 98% (103/105), respectively. The PDD test had a positive predictive value of 86% (61/71) and a negative predictive value of 88% (95/108), vs 96% (46/48) and 79% (103/131), respectively for WLC. There were no reports of significant complications after the procedure. CONCLUSION: PDD using hypericin shows promise, as it has a higher sensitivity but equivalent specificity than WLC. It can be used to detect flat lesions not seen on WLC. PDD testing is also well tolerated with minimal side-effects. PMID- 15892805 TI - Quantitative molecular urinary cytology by fluorescence in situ hybridization: a tool for tailoring surveillance of patients with superficial bladder cancer? AB - OBJECTIVE: To determine whether it is possible to stratify patients with superficial bladder cancer into low- and high-risk groups for tumour recurrence/progression based on the chromosomal pattern detected by fluorescence in situ hybridization (FISH) in one urine cytology specimen used for follow-up testing. PATIENTS AND METHODS: Voided urine samples from 47 consecutive patients with urinary tract neoplasms (13 with no history of urothelial malignancy and 34 under follow-up after complete transurethral resection of superficial urothelial carcinoma of the bladder) were evaluated by liquid-based cytology (ThinPrep(R), CYTYC Corp., Boxborough, MA, USA) and UroVysion FISH (Vysis-Abbott, Downers Grove, IL). RESULTS: Of the 34 patients under surveillance, the UroVysion test was negative in four, 17 had loss of 9p21 sequences either alone or combined with low-frequency trisomy/ies or tetrasomy/ies of chromosomes 3, 7 and 17 in single cells (low-risk FISH), and 13 also had complex aneusomies of the remaining chromosomes (high-risk FISH). One of the four FISH-negative neoplasms, four of the 17 low-risk FISH cases and five of the 11 informative high-risk FISH-positive patients developed recurrence. Progression occurred only in patients with high risk FISH results, showing high-frequency complex chromosomal polysomies (four of 11). CONCLUSION: The results from this pilot study indicate that the UroVysion FISH test may help to individually assess the clinical behaviour of superficial bladder cancer, based on the chromosomal pattern of exfoliated tumour cells in follow-up urinary cytology. It might be of use to identify those patients likely to progress at earlier and curable stages of disease, and lengthen the surveillance period in those with persistent or recurrent low-risk disease. PMID- 15892806 TI - Anatomical radical retropubic prostatectomy: 'curtain dissection' of the neurovascular bundle. AB - OBJECTIVE: To investigate the topographical relationship of the cavernosal nerves (CNs) to seminal vesicles, prostate, rhabdosphincter and urethra during the development of the prostate, and to use the resulting morphological data to modify the surgical technique of nerve-sparing radical prostatectomy. MATERIALS AND METHODS: The study included 29 male fetuses (gestational age 9-37 weeks) and eight adult specimens assessed anatomically and histologically. Using the plastination technique and anatomical dissection, the course of the CNs was investigated in all specimens. Based on these morphological results, the technique of dissecting the CNs during nerve-sparing radical retropubic prostatectomy was modified. RESULTS: During early fetal development the fibres of the CNs enclose the prostatic and membranous urethra dorsally and laterally. During the growth of the prostate, the CNs running along the prostate become displaced further anteriorly and spread, thus forming a concave shape (like a 'curtain') of the neurovascular bundles (NVB). Therefore, dissection of the NVB has to start anteriorly to preserve all the nerve fibres that are spread along the surface of the lateral lobes of the prostate. CONCLUSIONS: From these anatomical findings we propose a modified 'curtain dissection' to improve preservation of the CNs running in the NVB, in which the incision of the periprostatic fascia and dissection of the NVB is far more anterior than previously described. PMID- 15892807 TI - Imaging with radiolabelled monoclonal antibody (MUJ591) to prostate-specific membrane antigen in staging of clinically localized prostatic carcinoma: comparison with clinical, surgical and histological staging. AB - OBJECTIVE: To evaluate the reliability of prostate scintigraphy using a radiolabelled antibody (MUJ591) raised against the external domain of prostate specific membrane antigen (PSMA) in the staging of early prostate cancer. PATIENTS AND METHODS: This was a prospective study of 16 patients who had radical retropubic prostatectomies (median PSA 9.75 ng/mL). All patients underwent PSMA imaging using MUJ591 radiolabelled with (99m)Tc using a photo-reduction technique. RESULTS: The findings of prostate imaging and histology were identical in seven patients. Scans showed understaging and overstaging in six and three patients, respectively. CONCLUSIONS: PSMA scintigraphy using (99m)Tc-labelled MUJ591 identifies the presence of prostate cancer, but is not sensitive in delineating micro-invasion of the capsule, seminal vesicles or bladder neck. As in other studies it seems to be useful in detecting prostate bed recurrence and distant micrometastasis. PMID- 15892808 TI - Diverse optical characteristic of the prostate and light delivery system: implications for computer modelling of prostatic photodynamic therapy. AB - OBJECTIVE: To explore the use of photodynamic therapy (PDT) as a minimally invasive form of treatment for organ-confined prostate cancer, for although there are several therapies, ablative treatments are associated with significant morbidity. MATERIALS AND METHODS: Using the photosensitizer tin etiopurpurin, dogs were treated with interstitially placed laser fibres in an effort to validate PDT for treating prostate cancer. Earlier models assumed a uniform distribution of light output from a cylindrical fibre and a uniform attenuation coefficient throughout the prostate. Subsequent observations show that this model was too simple and that light radiance is not linear. To overcome under treatment, a computer program to complement real-time fibre placement was developed. RESULTS: As light radiance from interstitially placed laser fibres varies significantly from the commonly assumed ideal cylindrical emission, a predictive mathematical model of prostate PDT needs to consider the real emission. Also, the optical properties of the prostate, e.g. absorption and scattering of light, are anisotropic. Differences in the attenuation coefficient (combining absorption and scattering of light) also varied among different animals. Incorporating all these variables into a computer program produced a virtual model of the photo-ablated zone within +/- 2 mm of that observed in animals. CONCLUSION: PDT of the prostate is not trivial and should benefit from computer-aided methods as it is developed for clinical use. PMID- 15892809 TI - Volume-adjusted prostate-specific antigen (PSA) variables in detecting impalpable prostate cancer in men with PSA levels of 2-4 ng/mL: transabdominal measurement makes a significant contribution. AB - OBJECTIVE: To examine whether prostate-specific antigen (PSA) levels adjusted according to prostate volume improve prostate cancer detection using transrected biopsies in men with PSA levels of 2-4 ng/mL, and benign findings on a digital rectal examination (DRE). PATIENTS AND METHODS: Men aged < or = 79 years and with serum PSA levels of 2-4 ng/mL and normal DRE findings were prospectively enrolled. Eligible patients were recommended for transrectal prostate biopsies after measuring prostate volumes with transrectal (TRUS) and transabdominal (TAUS) ultrasonography, and transition zone volumes with TRUS. In addition to PSA levels and the free-to-total PSA ratio, volume-adjusted PSA levels, PSA densities determined by TRUS (PSAD(TRUS)), and TAUS (PSAD(TAUS)), and PSA transition zone densities (PSATzD) were compared using receiver operating characteristic analysis. RESULTS: Prostate cancer was diagnosed in 31 (22%) of the 139 men who had prostate biopsies. The area under the curve (AUC) of PSAD(TRUS) (0.796) and PSATzD (0.792) was similar and significantly greater than that of PSA (AUC 0.588) and the free-to-total PSA ratio (AUC 0.658). PSAD(TAUS) was a significantly better indicator of prostate cancer than PSA levels alone (P = 0.043). CONCLUSION: As predictors of prostate cancer, there were no significant differences between PSAD(TRUS) and PSATzD. Although PSAD(TAUS) was worse than PSA variables adjusted by total and transition zone prostate volumes determined by TRUS, it was a better predictor than the PSA value alone in men with a low PSA level. These results indicate that TAUS is worthwhile where the routine use of TRUS before biopsy is difficult. PMID- 15892810 TI - Prostate cancer detection in men with a 'normal' total prostate-specific antigen (PSA) level using percentage free PSA: a prospective screening study. AB - OBJECTIVE: To assess the utility of percentage free/total prostate-specific antigen (f/tPSA) levels for detecting prostate cancer in a prospectively screened population of men with a 'normal' total PSA level. PATIENTS AND METHODS: Men aged 50-65 years were contacted via their general practitioner and invited for prostate cancer screening. All had their total and f/tPSA levels measured; those meeting the biopsy criteria (PSA 1.1-3.99 ng/mL and f/tPSA < or = 20%) were offered a biopsy. The cancer detection rate was then evaluated and compared with other methods of detection. In all, 773 men were screened, of whom 115 met the criteria and agreed to undergo a prostate biopsy. RESULTS: Cancer was detected in 13 of the 115 men (11.3%) of whom most would have been missed by lowering the age adjusted threshold for total PSA to 2.5 ng/mL. There was no significant difference in total and f/tPSA values in men with and without prostate cancer. Those cancers that could be evaluated were found to be clinically significant. CONCLUSION: In this study prostate cancer was detected solely on the basis of a low f/tPSA value. Most men with cancer would have been missed by simply lowering the age-adjusted threshold for total PSA. Using the f/tPSA level may allow the detection of clinically significant cancer in men at a time when they are most likely to benefit from treatment. PMID- 15892811 TI - A novel surgical technique for implanting a new electrostimulation system for treating female overactive bladder: a preliminary report. AB - OBJECTIVE: To assess the results of peripheral electrostimulation of pelvic floor muscles in patients with overactive bladder (OAB) symptoms, and to describe a novel surgical technique for inserting a specially designed implantable electrostimulator which can deliver different forms of muscle stimulation through a para-urethral electrode. PATIENTS AND METHODS: The study included seven women with OAB (mean age 59 years) who had frequency (including nocturia), urgency and urge incontinence for > or = 1 year, and in whom conventional treatment failed. The antepubic technique was used to implant the electrostimulator and a para urethral electrode. The patients' urinary symptoms and quality of life (King's College Hospital incontinence questionnaire) were compared before and after surgery, and possible complications monitored. RESULTS: The mean (range) follow up was 14.5 (13-17) months in six and 10 months in the seventh patient (stimulator removed because of infection). The mean (range) frequency of 15 (13 23) voids/24 h before surgery decreased to 9 voids/24 h (i.e. by 40%) afterward. All patients had leakage episodes because of urgency (4-15 times/day) before surgery, which ceased in five afterward; these patients used no further pads. In the remaining two patients urgency incontinence continued after surgery, but with significantly fewer leakage episodes, from 15 and 12 per day before to 6.7 and 4 afterward, respectively. The mean degree of urgency (graded 0-3) decreased from 2 to 1.4 after surgery, and quality of life improved significantly. Urodynamic studies showed a greater mean maximum detrusor capacity, from 135 mL before to 189 mL after implanting the stimulator. CONCLUSIONS: The good results (over a follow-up of 1 year) in patients with OAB symptoms treated by the present electrostimulator should encourage further clinical studies in such patients. A trial to evaluate the efficacy of pelvic floor muscle electrostimulation in a broad spectrum of diseases of the lower urinary tract causing symptoms similar to OAB has been started. PMID- 15892812 TI - Classification and distribution of symptomatic nocturia with special attention to duration of time in bed: a patient-based study. AB - OBJECTIVE: To classify the night-time urinary frequency of patients with symptomatic nocturia by urine volume produced, nocturnal bladder capacity and time spent in bed. PATIENTS, SUBJECTS AND METHODS: In all, 110 patients whose principal complaint was nocturia were enrolled in the study; 32 age- and gender matched subjects were recruited as controls. From studies of the controls, 190 mL and 510 min were identified as the normal limits for mean nocturnal urine voided volume and time spent in bed. The 110 patients were classified into eight categories based on urine volume, nocturnal bladder capacity and time spent in bed. RESULTS: There were abnormalities of urine volume produced, bladder capacity and time spent in bed in 72%, 50% and 54% of the patients, respectively. Patients with severe nocturia (> or =4 voids/night) tended to have more complex pathophysiology than those with mild nocturia (< or =3 voids/night). CONCLUSION: Time spent in bed, urine volume produced and bladder capacity should be used to assess patients with symptomatic nocturia. PMID- 15892813 TI - Congenital adrenal hyperplasia and lower urinary tract symptoms. AB - OBJECTIVES: To assess urinary symptoms in adult women with congenital adrenal hyperplasia (CAH), as feminizing surgery in infancy is current standard practice for CAH and one of the indications for surgery is to reduce urinary symptoms. PATIENTS, SUBJECTS AND METHODS: In a case-control study, 19 women with CAH, of whom 16 had had childhood feminizing genital surgery, and age-matched women with no CAH, were evaluated. Subjects and controls completed the Bristol Female Lower Urinary Tract Symptoms (BFLUTS) questionnaire. RESULTS: Urge incontinence was reported in 13 (68%) patients and three (16%) controls (P = 0.003); stress incontinence was present in 47% and 26%, respectively (P = 0.31). Results from the controls were comparable with those documented in larger studies on normal populations. Nine of the patients felt that their urinary symptoms had an adverse effect on their lives, compared with only one of the controls (P = 0.008). CONCLUSION: Patients with a diagnosis of CAH are more likely to have significant urinary symptoms than normal controls. At present it is not clear whether this is a result of surgery or an effect of CAH. In at least two-thirds of patients surgery did not achieve the objective of reducing urinary symptoms. PMID- 15892814 TI - Is laparoscopic unilateral sural nerve grafting during radical prostatectomy effective in retaining sexual potency? AB - OBJECTIVES: To present a pilot study of laparoscopic unilateral sural nerve grafting during radical prostatectomy, with the aim of preserving sexual potency. PATIENTS AND METHODS: Because they had localized prostate cancer, 29 men had a laparoscopic radical prostatectomy with deliberate wide unilateral neurovascular bundle resection and preservation of the contralateral bundle. Fifteen men (group A) had an interposition sural nerve graft on the sectioned bundle, and 14 (group B) had laparoscopic radical prostatectomy with preservation of the unilateral bundle only. The men were also involved in a rehabilitation programme, and erectile function was evaluated after surgery, and at 3, 8, 12 and 18 months, using the five-item version of the International Index of Erectile Function (IIEF 5) questionnaire. RESULTS: The two groups had similar clinical characteristics (age, prostate-specific antigen level, body mass index, prostate volume, clinical stage, Gleason score before and after surgery, postoperative stage). The follow up was complete for 12 men in group A and 10 in group B. Group A had significantly higher erectile function scores on the IIEF-5 at 12 and 18 months than immediately after surgery (P < 0.01), whereas in group B the improvement was not statistically significant. Overall, by 18 months after surgery five of 12 men in group A had achieved spontaneous unassisted erection or erection assisted with sildenafil, while three of 10 in group B achieved an erection assisted with sildenafil (not significant). CONCLUSIONS: These data suggests that laparoscopic sural nerve grafting during radical prostatectomy is feasible and safe; nevertheless we cannot conclude that sural nerve grafting is more effective than preserving the neurovascular bundle alone in retaining sexual potency. More research is required to validate the effectiveness of this technique. PMID- 15892815 TI - Does thrombin sealant allow nephron-sparing surgery with no renal artery occlusion? A description of technique and initial results. PMID- 15892816 TI - Validation of computer-based training in ureterorenoscopy. AB - OBJECTIVES: To evaluate the outcome of training both urological novices and experts, using the recently developed UroMentor (Simbionix Ltd, Israel) trainer, that provides a realistic simulation of rigid and flexible ureterorenoscopy (URS). SUBJECTS AND METHODS: Twenty experienced urologists (total number of previous flexible URSs 21-153) were monitored during simulated flexible URS for treating a lower calyceal stone, and the outcome was correlated with individual experience. A score was compiled based on the variables recorded, including total operation time, stone contact time, complications such as bleeding or perforation, and treatment success. A further five urological residents with no endourological experience were trained on the UroMentor in rigid URS for ureteric stone treatment. Their acquired clinical skills were subsequently compared to those of five urological residents who received no simulator training. RESULTS: All 20 experienced urologists disintegrated the stone on the simulator, and the score achieved was related to their personal experience; there was a significant difference in performance in those with < 40 and > 80 previous flexible URSs. For the five urological residents with no endourological experience, simulator training improved their skills, and comparison with urological residents who had received no simulator training showed advantages for the trained residents. After being trained on the simulator, the group performed better in the first four URSs on patients. CONCLUSIONS: Individual experience correlates with individual performance on the simulator. Simulator training was helpful in improving clinical skills. Although the distribution of computer-based simulators is limited by high prices, virtual reality-based training has the potential to become an important tool for clinical education. PMID- 15892817 TI - Planning percutaneous nephrolithotomy using multidetector computed tomography urography, multiplanar reconstruction and three-dimensional reformatting. AB - OBJECTIVE: To assess a modified technique of multidetector computed tomographic urography (CTU) which can reproducibly and accurately map the pelvicalyceal system (PCS) and complex renal calculi, as such information is essential in choosing the optimal percutaneous approach into the PCS for safe and successful percutaneous nephrolithotomy (PCNL). PATIENTS AND METHODS: Ten consecutive patients with renal calculi underwent a modified four-detector multislice CTU with frusemide, abdominal compression and subsequent injection with contrast medium. After unenhanced CT of the abdomen, a high-resolution contrast-enhanced scan was taken through the kidneys in the pyelographic phase. Data were analysed using multiplanar reconstruction and three-dimensional (3D) reformatting. RESULTS: In 10 CTUs there were three staghorn, two diverticular, 25 calyceal, two infundibular and two renal pelvic calculi; nine showed posterior calyces and good infundibular anatomy, and provided a good map of the PCS. Seven patients had PCNL, with the remaining three having either primary extracorporeal shock wave lithotripsy or conservative management. CTU detected stones in all patients and accurately located their relation to the PCS. With reconstructed images, subjectively the 3D imaging provided an advantage over conventional imaging in optimizing nephrostomy placement. CONCLUSION: CTU with this protocol and post processing techniques enables an accurate and confident, reproducible prediction of the site, number and size of stones in complex pelvicalyceal anatomy, optimal site(s) for placing the percutaneous track, and potential hazards when placing the track, with no significant increase in the patient's radiation burden. 3D CTU should become the standard imaging method for planning PCNL in selected patients. PMID- 15892818 TI - Role of computed tomography with no contrast medium enhancement in predicting the outcome of extracorporeal shock wave lithotripsy for urinary calculi. AB - OBJECTIVE: To evaluate the usefulness of urinary calculi attenuation values from non-contrast computed tomography (NCCT) in predicting the outcome of treatment by extracorporeal shock wave lithotripsy (ESWL). PATIENTS AND METHODS: We evaluated 112 patients with solitary renal and upper ureteric calculi of 0.5-2 cm undergoing ESWL. All patients had NCCT at 120 kV and 240 mA on a spiral CT scanner. During each ESWL session 3000 shock waves were given to a maximum of 3.0 kV. A final X-ray of the kidney, ureters and bladder was taken 12 weeks after the last ESWL session. Fragments of < or = 5 mm were regarded as clinically insignificant residual fragments (CIRF). The calculi retrieved were analysed by X ray diffraction and the results assessed by comparing the mean density (as measured in Hounsfield units, HU) with the number of ESWL sessions and clearance. RESULTS: In all, 82 (76%) patients had complete clearance of stones and 26 (24%) had CIRF. There was a linear relationship between the calculus density and number of ESWL sessions required. Of patients with calculi of < or = 750 HU, 41 (80%) needed three or fewer ESWL sessions and 45 (88%) had complete clearance. Of patients with calculi of > 750 HU, 41 (72%) required three or more ESWL sessions, and 37 (65%) had complete clearance. The best outcome was in patients with calculus diameters of < 1.1 cm and mean densities of < or = 750 HU; 34 (83%) needed three or fewer ESWL sessions, and the clearance rate was 90%. The worst outcome was in patients with calculus densities of > 750 HU and diameters of > 1.1 cm; 23 (77%) needed three or more ESWL sessions and the clearance rate was only 60%. The calculus density was a stronger predictor of outcome than size alone. CONCLUSIONS: The use of NCCT for determining the attenuation values of urinary calculi before ESWL might help to predict the treatment outcome, and so might help in planning alternative treatment in patients with a likelihood of a poor outcome from ESWL. PMID- 15892819 TI - Postoperative resorptive and excretory capacity of the ileal neobladder. AB - OBJECTIVE: To assess the extent and mechanism of renal reabsorption and excretion in patients with an ileal neobladder, as mild metabolic acidosis cause by proton reabsorption is common after such surgery, and long-term pharmacological correction is often necessary. PATIENTS AND METHODS: The study comprised 30 patients (29 men and one woman) with ileal neobladders after oncological surgery; before surgery all had normal retention values. Before and after withdrawing the transurethral catheter, serum creatinine and urea were analysed and used to assess the effect of the neobladder on retention values, expressed as the percentage change from baseline (Delta creatinine and urea). RESULTS: There was a significant correlation between the Delta-creatinine and Delta-urea values (P < 0.001; r = 0.66); 15 patients (50%) showed resorption of creatinine and urea, eight (27%) excreted creatinine into the neobladder and resorbed urea from it at the same time, and three (10%) showed the reverse response, i.e. creatinine resorption and urea excretion. Interestingly, four patients (13%) excreted both creatinine and urea into the neobladder. CONCLUSIONS: We assume that there was both a resorptive and excretory function. Probably the metabolic state (resorption or secretion) of the neobladder depends on its mucus production and on the internal surface, or on diuresis. Further investigation is required to characterize these different influences. PMID- 15892820 TI - The pitfalls of vaginal construction. AB - OBJECTIVE: To evaluate a group of young women undergoing complex vaginoplasty, which may be defined as where there has already been previous complex pelvic surgery and the patient is unsuitable for conservative treatment or minor surgical revision. PATIENTS AND METHODS: All patients who had undergone a complex vaginoplasty between 1997 and 2004 were reviewed retrospectively. Information was recorded on the underlying condition, previous surgery and preoperative assessment. The operative technique and complications were noted, and the outcome in terms of menstruation and sexual activity. In all, 15 patients fulfilled the criteria (mean age 22.2 years, range 17-36) and the mean follow-up was 4.75 (0.75 7) years. RESULTS: Fourteen patients had complex congenital anomalies of the lower genital tract and 11 of these had associated anomalies of the urinary and gastrointestinal system; four were XY females. The 15 patients had had 31 vaginal procedures, including ileal vaginoplasty, mobilization of perineal skin flaps and split-skin grafting. Three patients required osteotomies to increase pelvic outlet diameter. The commonest complications were stenosis (six patients) and fistula formation (four). Eight patients are now sexually active; the remaining seven have not attempted sexual intercourse. CONCLUSIONS: Complications are common and can be major; most patients require many repeat operations to achieve a patent vagina. Unfortunately at present, this is the only option for this group of young women to achieve a functional vagina. Patients with complex congenital anomalies of the reproductive and genital tracts are increasingly surviving into adulthood, and the numbers of these patients is likely to increase in the future. PMID- 15892821 TI - A comparison of one-stage procedures for post-traumatic urethral stricture repair. AB - OBJECTIVE: To compare the results and complication rates of various one-stage treatments for repairing a post-traumatic urethral stricture. PATIENTS AND METHODS: The medical records of 153 patients who had a post-traumatic urethral stricture repaired between 1977 and 2003 were evaluated retrospectively, and analysed for the different types of urethral reconstruction. RESULTS: The procedures included direct end-to-end anastomosis in 86 (56%) patients, free dorsal onlay graft urethroplasty using preputial or inguinal skin in 40 (26%), ventral onlay urethroplasty using buccal mucosa in seven (5%) and ventral fasciocutaneous flaps on a vascular pedicle in 20 (13%). At a mean (median, range) follow-up of 75.2 (38, 12-322) months, 121 (79%) patients had no evidence of recurrent stricture, while in 32 men (21%) they were detected at a mean follow up of 30.47 (1-96) months. Patients having a dorsal onlay urethroplasty had the longest strictures. The re-stricture rate was lowest after a dorsal onlay urethroplasty (5% vs 27% when treated with end-to-end anastomosis, 15% after fasciocutaneous flaps and 57% after a ventral buccal mucosal graft). The surgical technique used had no effect on postoperative incontinence or erectile dysfunction rates. CONCLUSION: In patients with strictures which are too long to be excised and re-anastomosed, tension-free dorsal onlay urethroplasty is better than ventral graft or flap techniques. In patients with short urethral strictures direct end-to-end anastomosis remains an option for the one-stage repair of urethral stricture. PMID- 15892822 TI - Dorsal dartos flap for preventing fistula in the Snodgrass hypospadias repair. AB - OBJECTIVE: To evaluate the importance of urethral covering using vascularized dorsal subcutaneous tissue for preventing fistula in the Snodgrass hypospadias repair. PATIENTS AND METHODS: The study included 67 children (aged 1-11 years) who had hypospadias repaired between April 1998 and May 2003, including 51 with distal and 16 with midshaft hypospadias. In all children, a standard tubularized incised-plate urethroplasty was followed by reconstruction of new surrounding urethral tissue. A longitudinal dartos flap was harvested from excessive dorsal preputial and penile hypospadiac skin, and transposed to the ventral side by a buttonhole manoeuvre; it was sutured to the glans wings around the neomeatus, and to the corpora cavernosa over the neourethra. Thus the neourethra was completely covered with well-vascularized subcutaneous tissue. RESULTS: At a mean (range) follow-up of 21 (6-65) months, the result was successful, with no fistula or urethral stenosis, in all 67 children. CONCLUSION: We suggest that urethral covering should be part of the Snodgrass procedure. A dorsal well-vascularized dartos flap, buttonholed ventrally, is a good choice for preventing fistula. Redundancy of the flap and its excellent vascularization depends on the harvesting technique. PMID- 15892823 TI - Extracorporeal pelvic floor magnetic stimulation in children with voiding dysfunction. AB - OBJECTIVE: To determine the effect of extracorporeal pelvic floor magnetic stimulation in children with an overactive bladder, as although such stimulation is an effective treatment for voiding dysfunction such as urge incontinence (UI) and urgency-frequency syndrome, experience in children is scarce. PATIENTS AND METHODS: This prospective study included 42 children diagnosed with an overactive bladder, based on urodynamic or video-urodynamic study; a complete follow-up was available in 34. The children were grouped into those with UI only, not monosymptomatic nocturnal enuresis (nMNE), or MNE, according to their symptoms. Clinical variables were assessed by recording a voiding and nocturnal enuresis diary before and after magnetic stimulation, the latter being administered twice a week for 4 weeks using a size-adjusted magnetic chair (each session took 20 min). RESULTS: The UI only and nMNE group had a significant decrease in voiding frequency and frequency of UI (P < 0.05); the MNE group also had a significant decrease in voiding frequency (P < 0.05). There was a significant increase in functional bladder capacity in all groups (P < 0.05) but no significant decrease in the mean volume and frequency of NE in the nMNE and MNE groups (P > 0.05). CONCLUSIONS: Extracorporeal pelvic floor magnetic stimulation has an acute effect on voiding dysfunction such as urge syndrome in children. However, controlled studies with a sham-stimulation group and various durations of stimulation are necessary for its application as a primary treatment for voiding dysfunction in children. PMID- 15892824 TI - The effect of location of the ureteric orifice on the efficacy of endoscopic injection to correct vesico-ureteric reflux. AB - OBJECTIVE: To review our 11-year experience and identify the mechanisms responsible for the failure of endoscopic injection for vesico-ureteric reflux (VUR) with three different injectable agents, based on the location of the ureteric orifice on endoscopy. PATIENTS AND METHODS: We retrospectively reviewed the charts and endoscopic video-photographs of 46 patients (26 girls, 20 boys, median age 6 years, range 2-16) with VUR treated once or twice by subureteric injection with PTFE, or polydimethylsiloxane or dextranomer/hyaluronic acid copolymer, from 1992 to 2003. Five patients were lost to follow-up and six ectopic and/or duplicated ureters were excluded from the analysis; in all, 52 ureters were analysed. According to the international classification, the VUR was grades I to V in four (8%), 12 (23%), 16 (31%), 13 (25%) and seven (13%) ureters, respectively. RESULTS: After 3 months, voiding cysto-urethrography showed that VUR continued in six of 19, seven of 12 and eight of 21 ureters (38%), respectively, after subureteric PTFE, polydimethylsiloxane and dextranomer/hyaluronic acid copolymer injection; after the second injection, reflux continued in two of six, four of seven and three of eight ureters, respectively. Mound displacement and/or volume loss was the most common failure with all three bulking agents after both the first (62%) and second injections (44%) (P < 0.05). The first injection failed in 32% (11 of 35) normally located ureters and 10 of 17 lateral ureters (P < 0.05). The second injection failed in 11% (four of 35) normal and five of 17 lateral ureters (P < 0.05). CONCLUSIONS: A lateral ureteric orifice may decrease the efficacy of endoscopic injection, as the likelihood of a faulty injection is greater. However, a more careful second injection decreases the failure rate, particularly in those with low- to medium grade refluxing ureters. PMID- 15892825 TI - The androgen receptor and signal-transduction pathways in hormone-refractory prostate cancer. Part 1: Modifications to the androgen receptor. PMID- 15892826 TI - The androgen receptor and signal-transduction pathways in hormone-refractory prostate cancer. Part 2: Androgen-receptor cofactors and bypass pathways. PMID- 15892827 TI - Immunotherapy of murine prostate cancer using whole tumour cells killed ex vivo by cytosine deaminase/5-fluorocytosine suicide-gene therapy. AB - OBJECTIVE: To evaluate the efficacy of antitumour vaccines comprising irradiated allogeneic or autologous whole cells expressing cytosine deaminase (CD) which are first killed ex vivo by prodrug activation using 5-fluorocytosine (5-FC), as the immunogenicity of tumour cells used as irradiated vaccines depends both on antigen expression and on the mode of their death. MATERIALS AND METHODS: The PA3 rat prostate cell line and MATLyLu, an androgen-insensitive subline, were grown and transfected with CD (designated PCD and MCD). In vitro drug-sensitivity was assessed in the cell lines using a viability assay, and the mode of cell death quantified by assessing apoptosis. Bax and bcl-2 expression were assessed by Western blot analysis. For in vivo experiments, male 8-10-week-old Lobund-Wistar rats were vaccinated (using vehicle in control groups) with 5 x 10(6) cells, all cells being irradiated before injection, to give groups with PA3, PCD, PCD killed with 5-FC, MatLyLu, MCD, and MCD killed with 5-FC. After 7 days all animals were given a subcutaneous tumour challenge of PA3 cells, and tumour volume measured subsequently. Immune responses were assessed in splenocytes. RESULTS: The efficiency of cell kill varied between the cell lines assessed, but cell death was by induced apoptosis. Single doses of vaccine were most effective in the allogeneic setting, causing significantly slower growth of syngeneic tumour challenge (P < 0.01), and 25% better survival at 50 days (P < 0.02) than irradiated untransfected cells. This was consistent with the greater proliferative response after allogeneic than autologous vaccination. CONCLUSION: The immunogenicity of irradiated tumour cells is enhanced when they are killed ex vivo using suicide-gene therapy. This approach would be clinically applicable in terms of ease of vaccine production, safety, storage and avoidance of potential toxicities of in vivo gene transfer. PMID- 15892828 TI - Expression of the epidermal growth factor receptor family in normal and malignant urothelium. AB - OBJECTIVE: To compare the immunohistochemically assessed expression of the epidermal growth factor receptor (EGFR) family in normal and malignant bladder urothelium, and suggest new hypotheses about their function in the development and progression of transitional cell carcinoma (TCC). PATIENTS AND METHODS: EGFR, ERBB2, ERBB3 and ERBB4 were evaluated immunohistochemically in normal urothelium (NU, 15), primary non-metastasized invasive TCC (NMC, 19) and in primary invasive TCCs with corresponding metastases (MC, 51, both specimens). RESULTS: All NU samples expressed ERBB4, none expressed ERBB2 and two expressed EGFR; all staining was uniform throughout all cell layers. ERBB2 expression increased and ERBB4 decreased from normal samples to carcinomas. There was no difference between NMCs and MCs in ERBB2, ERBB3 and ERBB4, but the NMCs expressed more EGFR than both NU and MC samples. There were no associations with T category, grade or survival. All combinations of expression levels for the four receptors were detected, with no dominant profile. CONCLUSION: We hypothesise that: (i) ERBB4 is important for differentiation in NU; (ii) ERBB2 is up-regulated with carcinogenesis in the urinary bladder but does not discriminate between bladder cancer with or without metastases; (iii) EGFR may be a marker of indolent disease. A current hypothesis, that superficial layers of NU do not express EGFR and thus protect the basal cells from the mitogenic effect of urinary EGF, is challenged. PMID- 15892829 TI - In vitro viability of human cavernosal endothelial and fibroblastic cells after exposure to papaverine/phentolamine and prostaglandin E1. AB - OBJECTIVE: To investigate the influence of commercially available vasoactive drugs on human cavernosal endothelial and fibroblastic cells in vitro, as although corporal fibrosis is a well known side-effect of intracavernosal injection therapy for erectile dysfunction, the possible detrimental effect of these agents on the endothelium lining the cavernosal vascular spaces is uncertain. MATERIALS AND METHODS: Cultured primary endothelial (13) and fibroblastic cells (12), obtained from potent patients undergoing penile surgery, were exposed to different physiological dilutions of prostaglandin E1 (PGE1), papaverine/phentolamine or the respective triple-mix of these agents for 30 min. Viable cells were counted and cell metabolic activity measured in these cultures 48 h after drug exposure. RESULTS: There was a significant dose-dependent decrease in the viable cell count after exposure to papaverine-containing formulations, probably because of the low pH of this substance. This cytotoxic effect was more pronounced in endothelial than in fibroblastic cells, and was not apparent in the PGE1 groups. The relative increase in cell metabolic activity in cultures affected by a moderate cytotoxic effect indicated a regenerative process. CONCLUSION: These comparative results in endothelial and fibroblastic cell cultures suggest that the endothelium rather than the interstitium of the corpus cavernosum is more sensitive to side-effects produced by intracavernosal injection therapy with papaverine. Thus, unfavourable consequences on the function of the endothelial layer might be as important as the risk of interstitial fibrosis. As these effects were not detected for PGE1 this drug should be preferred to papaverine in clinical practice. PMID- 15892830 TI - Evergreening: there's life in the old drug yet. PMID- 15892831 TI - A simple and comfortable technique for replacing ureteric stents in patients with strictured uretero-intestinal anastomosis or strictures at the vesico-ureteric junction. PMID- 15892832 TI - The self-anchoring transobturator male sling to treat stress urinary incontinence in men: a new sling, a surgical approach and anatomical findings in a cadaveric study. PMID- 15892834 TI - Mini-reviews. PMID- 15892833 TI - Re: Managing patients with an overactive bladder and glaucoma: a questionnaire survey of Japanese urologists on the use of anticholinergics. PMID- 15892835 TI - Re: Where are the 'poster boys' for bladder cancer? PMID- 15892836 TI - Re: Re-evaluation of the tumour-node-metastasis staging of locally advanced renal cortical tumours: absolute size (T2) is more significant than renal capsular invasion (T3a). PMID- 15892837 TI - Re: Examining the location and the cause of death within 30 days of radical prostatectomy. PMID- 15892838 TI - Re: Diffusion properties of transurethral intraprostatic injections. PMID- 15892840 TI - Tumour nephrectomy with vena cava thrombus. PMID- 15892843 TI - [Viewpoint of the German Dermatologic Society (DDG) concerning the decision of the American Food and Drug Administration (FDA) on the use of pimecrolimus cream and tacrolimus ointment in the treatment of atopic dermatitis (neurodermatitis)]. PMID- 15892844 TI - [Malignant melanoma -- a medical students' viewpoint]. AB - Malignant melanoma is a primarily cutaneous melanocytic tumour with increasing incidence responsible for 90 % of skin cancer mortality. Genetic predisposition has been identified as the most important risk factor, while UV is second in importance and can be avoided. New diagnostic methods include sentinel lymph node biopsy and the detection of tumour markers in blood. Furthermore, malignant melanoma shows an extraordinary resistance to therapy; at present the only cure lies in early excision of the primary tumour. Thus early recognition is of utmost importance. Experimental approaches, such as dendritic cell vaccination, have shown some effectiveness which must be confirmed in multicenter, randomised trials. PMID- 15892845 TI - [Significant decrease in quality of life in patients with pemphigus vulgaris. Results from the German Bullous Skin Disease (BSD) Study Group]. AB - BACKGROUND: Pemphigus vulgaris is a potentially life-threatening autoimmune disorder of the skin and mucous membranes characterized by antibodies against epidermal adhesion molecules. Clinically characteristic are painful chronic blisters or erosions of mucous membranes and skin. There are no published studies on the impact o this disease on quality of life. PATIENTS AND METHODS: This registration was performed within the scope of the German BSD (Bullous Skin Disease) study group, from November 1997 until January 2002. A total of 36 patients with the first diagnosis of pemphigus vulgaris were registered at the university hospitals of Dresden, Erlangen, Kiel, Mannheim, Munchen and Wurzburg. Thirty of the 36 (83 %) patients participated in the quality of life questionnaire utilizing the German version of 'Dermatology Life Quality Index' (DLQI) provided by A. Y. Finlay. The DLQI varies from 0 to 30 with an increased DLQI score indicating a decrease in quality of quality. RESULTS: The overall DLQI total score of 10 +/- 6.7 in the investigated pemphigus patients was significantly increased in comparison to other skin diseases. CONCLUSIONS: These results suggest that the DLQI can be a very useful additional outcome criteria for clinical studies with pemphigus vulgaris and in the treatment of these patients. PMID- 15892846 TI - Neodymium-YAG Laser for hemangiomas and vascular malformations -- long term results. AB - BACKGROUND: Hemangiomas and vascular malformations are the most common vascular lesions of infancy. Different lasers can be used for treatment. Nd:YAG laser photocoagulation is particularly effective because of its deep penetration into tissue. PATIENTS AND METHODS: Thirty-one patients, aged from three months to 18 years, with voluminous hemangiomas and venous malformations were treated with a cw-neodymium:YAG laser. The quartz fibre was used in percutaneous and intralesional technique. Long-term follow-up data were acquired by clinical control or a patient questionnaire for a maximal period of eight years. Twenty patients could be evaluated. RESULTS: In the group with hemangiomas (n = 15), three cases showed nearly complete remission (> 90 %), ten cases had a partial reduction in size (50 - 90 %), in one case there was stable disease and in one case tumor growth. In the group with venous malformations (n = 5) two cases showed an excellent response (> 90 %), one case a moderate response (25 - 50 %) and in two cases there was no improvement. Adverse effects included scars (40 %), hyper- and hypopigmentation (23 %), mild atrophy (20 %) and a wrinkled texture (17 %). After maximal reduction in size, 30 % of the patients were not satisfied with the laser treatment outcome and elected surgical excision of the residual lesion. CONCLUSIONS: The neodymium:yttrium aluminium garnet (Nd:YAG) laser with percutaneous or intralesional application technique is a valuable tool for selected patients with hemangiomas and venous malformations. PMID- 15892847 TI - [Alopecia areata universalis and disseminated mollusca contagiosa in atopic dermatitis. Hair re-growth during treatment with interferon gamma--therapeutic effect or coincidence?]. AB - A 35-year-old woman presented with severe recalcitrant atopic dermatitis, in association with disseminated mollusca contagiosa and alopecia areata universalis. After several weeks of systemic interferon gamma, which was administered subcutaneously,the viral infection cleared and, surprisingly, four weeks after starting treatment hair re-growth was observed. Complete remission of alopecia areata was documented few weeks later and persists. After four cycles of high-dose intravenous immunoglobulin, a sustained remission of the atopic dermatitis was achieved. PMID- 15892848 TI - [Tumorous extragenital manifestation of lymphogranuloma venereum]. AB - A 36-year-old homosexual man presented with a 3-week history of pin-head-sized vesicles on the lower lip followed by a tumorous infiltrate, cervical lymphadenopathy and episodic fevers. Laboratory findings included striking leukocytosis and elevated inflammatory markers. Serologically,anti-chlamydial antibodies (IgG, IgA,IgM) were present. The histological findings were compatible with a chlamydial infection; PCR examination of the tumor and lymph nodes was positive for Chlamydia trachomatis DNA. Treatment with doxycycline (2x 100 mg/daily p.o. for 3 weeks) led to rapid improvement. Lymphogranuloma venereum is a relatively rare, worldwide illness, more common in tropical and sub-tropical zones. In Germany, there are 10-50 cases reported annually but many cases go unreported. The causative organism is Chlamydia trachomatis, serotypes L1-3. The transfer occurs through skin or mucous membrane contact with an infected partner. In the advanced stage, the infection can lead to marked internal organ involvement. The special features of our case include the extragenital manifestation and the tumorous appearance of the illness which is very rarely diagnosed in Germany. PMID- 15892849 TI - [Profiling methods in dermatology]. PMID- 15892850 TI - [Persistent facial erythema and edema]. PMID- 15892851 TI - [Therapy of psoriasis vulgaris and psoriatic arthritis with etanercept]. PMID- 15892852 TI - [Implementation and development of quality management in hospitals and private practice -- present state of the art]. AB - Since 1. 1. 2004 both hospitals and private practices in Germany are required by law to implement a quality management (QM) system within their organization. Different QM systems with a number of advantages and disadvantages are available. It has been well-documented that implementing QM im-proves the quality of both processes and results, thus improving patient care. At present certification of the organization is not yet part of the official requirements, but seems a likely step. PMID- 15892853 TI - Molecular genetics of hereditary prothrombin deficiency in Indian patients: identification of a novel Ala362 --> Thr (Prothrombin Vellore 1) mutation. AB - Prothrombin deficiency is a rare (1:200 000) autosomal recessive disorder caused by diverse mutations in prothrombin gene. We have studied the molecular basis of this disorder in four unrelated Indian patients. The diagnosis was based on prolonged prothrombin (PT) and activated partial thromboplastin times and low factor II coagulant activity (FII: C) measured using a PT based assay. FII: C levels ranged between 4.7% and 17.5%. Mutations were identified in all the four patients. Five different causative mutations including four (80%) missense and an in-frame deletion (20%) were identified. One of them was a novel, Ala362 --> Thr amino acid change affecting 'B' chain of -thrombin. This mutation was present in a compound heterozygous state with a previously reported Arg-1 --> Gln missense change affecting pro-peptide cleavage site. Ala362 --> Thr occurred at a codon, evolutionarily conserved in all the 24 different prothrombins or its related serine proteases studied. Molecular modeling of this mutation was found to cause a conformational change around the region involving a catalytic triad residue His363 and a cysteine residue at codon 364. The FII: C level in this patient was 17.5%. Three other previously reported mutations were also detected in the homozygous state: Arg271 --> Cys in Kringle-2 region, a Glu309 --> Lys in "A" chain of -thrombin and an in-frame deletion of 3 bp (AAG) leading to Del Lys301/302 in "A" chain of -thrombin. This is the first report of the molecular basis of prothrombin deficiency in Indian patients and we suggest the eponym 'Prothrombin Vellore 1' for Ala362 --> Thr mutation. PMID- 15892854 TI - Site and clinical outcome of deep vein thrombosis of the lower limbs: an epidemiological study. AB - Clinical outcomes of patients diagnosed with venous thromboembolic disease (VTED) have rarely been assessed on large series of patients from single institutions. This was work based on our practice to routinely screen all suspected pulmonary embolism (PE) and deep venous thrombosis (DVT) patients with bilateral proximal and distal venous US was designed to evaluate survival, recurrence and cancer occurrence in patients diagnosed with symptomatic or asymptomatic DVT and to assess their relationship with the site of the DVT (proximal vs. distal, bilateral vs. unilateral). Our study is based on the cross-matching of the VTED register of the Grenoble University Hospital with the local Cancer Register and community mortality data. Survival analyses were performed with the Kaplan-Meier method; prognostic variables were tested using the log-rank test. A total of 1913 patients with a DVT of the lower limbs from 1993 to 1998 were included (57% women; mean age, 69 years). Of these, 1018 patients were diagnosed with proximal DVT (156 bilateral) and 895 distal DVT (112 bilateral). PE was associated in 760 patients. Patients with PE and no detected DVT were not included. At 2 years, adjusted survival rates were 80% in patients with unilateral-distal DVT, 67% in bilateral-distal, 72% in unilateral-proximal and 65% in bilateral-proximal DVT patients. The cumulated VTED recurrence rates were 7.7% in unilateral-distal DVT, 13.3% when DVT was bilateral-distal, 14% when unilateral-proximal and 13.2% when bilateral-proximal. The rate of new cancer was 6.4% in unilateral-distal DVT, 10.8% when it was bilateral-distal, 6.5% when unilateral-proximal and 6.1% when bilateral-proximal. Based on a large series of unselected patients, our results show that the site of the DVT and principally the bilaterality provides important prognostic information that may be used in the setting up of medical strategies. PMID- 15892855 TI - The structure of thrombin, a chameleon-like proteinase. PMID- 15892856 TI - Factor XIII in bronchoalveolar lavage fluid from children with chronic bronchoalveolar inflammation. AB - BACKGROUND: Extravascular activation of the coagulation system and consequent fibrin deposition is involved in the pathomechanism of chronic bronchoalveolar inflammatory diseases. The turnover of extravascular fibrin is attenuated by its cross-linking with activated factor XIII (FXIII). OBJECTIVES: Determination of cellular and plasmatic forms of FXIII and their correlation with D-dimer level in the bronchoalveolar lavage fluid (BALF) from healthy children and from children with bronchoalveolar inflammation. PATIENTS AND METHODS: Highly sensitive immunoassays were used for the quantitation of cellular and plasma FXIII and D dimer in the BALF of children with recurrent wheezy bronchitis and fibrosing alveolitis. BALF was investigated for FXIII-containing cells by flow cytometry. RESULTS AND CONCLUSIONS: In the BALF of controls a low amount of the cellular form of FXIII (FXIII A2) and D-dimer were measured, while plasma FXIII (FXIII A2B2) was absent. Alveolar macrophages represented the single cell population in BALF that contained FXIII. In the BALF of both patients' groups the concentration and the total amount of FXIII A2 was significantly elevated, and plasma FXIII also appeared in the BALF of most patients. The D-dimer concentration was also elevated in the patients' groups and it correlated both with plasma FXIII and neutrophil count. These findings suggest that FXIII A2 is released from activated or injured alveolar macrophages into the bronchoalveolar lining fluid and in bronchoalveolar inflammatory diseases, FXIII A2B2 also leaks out from the capillaries. By cross-linking fibrin and inhibitors of fibrinolysis to fibrin, FXIII might be a key regulator of fibrin turnover in the extravascular compartment. PMID- 15892857 TI - Pharmacokinetics and pharmacodynamics of BB-10153, a thrombin-activatable plasminogen, in healthy volunteers. AB - BACKGROUND: BB-10153 is an engineered variant of human plasminogen that is activated to plasmin by thrombin. Thrombus-selective induction of reperfusion and prevention of reocclusion have been demonstrated following bolus administration in animal models of thrombosis. OBJECTIVE AND METHODS: The objective of the study was to examine the pharmacokinetics and pharmacodynamics of BB-10153 administered as an intravenous bolus to healthy male human volunteers. Cohorts of four were dosed with BB-10153 (n = 3) or placebo (n = 1). In total, placebo was received by eight volunteers and 0.08, 0.2, 0.6, 1.2, 1.8, 2.4, 3.6 and 4.8 mg kg(-1) BB 10153 by three volunteers each. RESULTS: There was a linear relationship between AUC/Cmax and dose. The half-life of BB-10153 was approximately 3-4 h and all the BB-10153 in the circulation retained the ability to be activated by thrombin. There was a dose-related increase in plasma fibrin D-dimers. Ex vivo plasma clot lysis was observed at doses of 3.6 and 4.8 mg kg(-1), whereas lysis of clots formed from euglobulin-fractionated plasma was first evident at 0.6 mg kg(-1) and activity increased with dose. This activity decreased with time in line with the half-life. BB-10153 had no effect on plasma alpha2-antiplasmin or fibrinogen levels, coagulation assays or bleeding time. An increase in plasminogen was observed as BB-10153 was detected by the enzyme-linked immunosorbent assay (ELISA) for human plasminogen. CONCLUSIONS: BB-10153 was well tolerated and had a 3-4-h plasma half-life. Fibrinolytic activity was demonstrated by dose-related ex vivo clot lysis and in vivo production of fibrin D-dimers. These effects were not accompanied by consumption of alpha2-antiplasmin or fibrinogen. PMID- 15892858 TI - Aspirin resistance: position paper of the Working Group on Aspirin Resistance. PMID- 15892859 TI - Thrombotic thrombocytopenic purpura and its diagnosis. PMID- 15892860 TI - Interspecies exchange mutagenesis of the first epidermal growth factor-like domain of human factor VII. AB - The first epidermal growth factor-like (EGF1) domain of human factor VII (FVII) is essential for binding to tissue factor (TF). We hypothesized that the previously observed increased coagulant activity of rabbit plasma (i.e. FVII) with human TF might be explained by the five non-conserved amino acids in the rabbit vs. the human FVII EGF1 domain. Accordingly, we 'rabbitized' the human FVII EGF1 domain either by exchanging the entire EGF1 domain creating human FVII(rabEGF1) or by the single amino acid substitutions S53N, K62E, P74A, A75D and T83K. After transient expression in HEK293 cells, the recombinant FVII (rFVII) mutant proteins were analyzed for biological activity and binding affinity to human TF by competitive enzyme-linked immunosorbent assay (ELISA). Biological activity of the unpurified rFVII mutant proteins was either depressed or statistically unchanged vs. rFVII(WT). However, three of six rFVII mutant proteins had increased affinity for human TF in the rank order rFVII(rabEGF1) (3.3-fold) > rFVII(K62E) (2.9-fold) > rFVII(A75D) (1.7-fold). The mutant protein rFVII(K62E) was then permanently expressed and purified. Fully activated, purified rFVIIa(K62E) had a twofold greater clotting activity and 2.8-fold greater direct FVIIa amidolytic activity when compared with rFVIIa(WT). Quantitation of the affinity of TF binding by surface plasmon resonance indicated that the KD of purified rFVII(K62E) for human soluble TF (sTF) was 1.5 nM compared with 7.5 nM for rFVII(WT), i.e. fivefold greater affinity. We conclude that substitution of selected amino acid residues of the FVII EGF1 domain facilitated the creation of human rFVII chimeric proteins with both enhanced biological activity and increased affinity for TF. PMID- 15892861 TI - A rare and misdiagnosed bleeding disorder: hereditary hemorrhagic telangiectasia. PMID- 15892862 TI - Atrial fibrillation in patients with first-ever stroke: frequency, antithrombotic treatment before the event and effect on clinical outcome. AB - BACKGROUND AND PURPOSES: Atrial fibrillation (AF) is an independent risk factor for stroke. The aims of this study were to assess: (i) the frequency of known or unknown AF in patients admitted to the hospital for a first-ever ischemic stroke and whether AF is associated with an adverse outcome at discharge (death or disability); (ii) the rates and determinants for the use of antithrombotic agents before stroke in patients with known AF and the adherence to the current treatment guidelines; and (iii) whether the lack of adherence to the current guidelines is associated with adverse outcome at discharge. METHODS: Consecutive patients with acute first-ever stroke admitted to an individual Stroke Unit between January 2000 to December 2003, were included in the study. Twelve-lead electrocardiogram (ECG) was performed in all patients on admission. Functional outcome was measured at discharge according to modified Rankin Score. RESULTS: A total of 1549 patients were included in the study: 238 patients (15.4%) were known to have AF and 76 (4.9%) were diagnosed with AF (unknown) on ECG performed on admission. At discharge 91 patients (5.9%) had died and 605 patients (39.0%) had died or were functionally dependent. Multivariate analysis showed that AF on admission was correlated with mortality or disability (OR = 1.58, 95% CI 1.09 2.30, P = 0.015). Before stroke, 124 out of 238 patients with known AF (52.1%) were not on antithrombotic therapy, 83 (34.9%) were receiving antiplatelet and 31 (13.0%) anticoagulant treatment. Previous transient ischemic attack, history of ischemic heart disease and hyperlipidemia were associated with the use of antithrombotic therapy. Only 24 out of 114 patients on antithrombotic treatment on admission were adequately treated according to the current guidelines. Of the adequately treated patients, 41.7% died or were disabled at discharge respect to 52.3% of the patients non-adequately treated (RR = 0.80, 95% CI 0.48-1.30). CONCLUSIONS: AF (on history or new diagnosis) was present in 20.3% of the patients with first-ever stroke admitted to a Stroke Unit and it was associated with increased mortality or disability. Only 10% of patients with known AF were previously receiving an adequate antithrombotic treatment according to current guidelines. PMID- 15892863 TI - Six novel mutations including triple heterozygosity for Phe31Ser, 514delT and 516T-->G factor X gene mutations are responsible for congenital factor X deficiency in patients of Nepali and Indian origin. AB - Factor X (FX) deficiency is a rare (1 : 100000) autosomal recessive disorder caused by heterogeneous mutations in FX gene. We have studied the molecular basis this disease in six Indian and one Nepali patients. Diagnosis was confirmed by measuring the FX coagulant activity (FX: C) using a PT based assay. Six of them had a FX: C of < 1% and one patient had 24% coagulant activity. Mutations were identified in all the seven patients. These included eight (88.8%) missense and one frame-shift (11.2%) mutations of which six were novel. Three of the novel mutations, a Phe31Ser affecting 'Gla' domain and 514delT and 516T-->G mutations affecting Cys132 in 'connecting region' were identified in a triple compound heterozygous state in a Nepali patient presenting with a severe phenotype. Two other novel mutations, Gly133Arg, may affect the disulphide bridge between Cys132 Cys302 in the connecting region while Gly223Arg may perturb the catalytic triad (His236, Asp282 and Ser379). The other novel mutation, Ser354Arg, involves the replacement of a small-buried residue by a large basic aminoacid and is likely to have steric or electrostatic effects in the pocket involving Lys351-Arg347-Lys414 that contributes to the core epitope of FXa for binding to FVa. Three previously reported mutations, Thr318Met; Gly323Ser; Gly366Ser were also identified. This is the first report of the molecular basis of FX deficiency in patients from the Indian subcontinent. PMID- 15892865 TI - Genetic factors contribute to bleeding after cardiac surgery. AB - BACKGROUND: Postoperative bleeding remains a common, serious problem for cardiac surgery patients, with striking inter-patient variability poorly explained by clinical, procedural, and biological markers. OBJECTIVE: We tested the hypothesis that genetic polymorphisms of coagulation proteins and platelet glycoproteins are associated with bleeding after cardiac surgery. PATIENTS/METHODS: Seven hundred and eighty patients undergoing aortocoronary surgery with cardiopulmonary bypass were studied. Clinical covariates previously associated with bleeding were recorded and DNA isolated from preoperative blood. Matrix Assisted Laser Desorption/Ionization, Time-Of-Flight (MALDI-TOF) mass spectroscopy or polymerase chain reaction were used for genotype analysis. Multivariable linear regression modeling, including all genetic main effects and two-way gene-gene interactions, related clinical and genetic predictors to bleeding from the thorax and mediastinum. RESULTS: Nineteen candidate polymorphisms were assessed; seven [GPIaIIa-52C>T and 807C>T, GPIb alpha 524C>T, tissue factor-603A>G, prothrombin 20210G>A, tissue factor pathway inhibitor-399C>T, and angiotensin converting enzyme (ACE) deletion/insertion] demonstrate significant association with bleeding (P < 0.01). Adding genetic to clinical predictors results improves the model, doubling overall ability to predict bleeding (P < 0.01). CONCLUSIONS: We identified seven genetic polymorphisms associated with bleeding after cardiac surgery. Genetic factors appear primarily independent of, and explain at least as much variation in bleeding as clinical covariates; combining genetic and clinical factors double our ability to predict bleeding after cardiac surgery. Accounting for genotype may be necessary when stratifying risk of bleeding after cardiac surgery. PMID- 15892866 TI - Spatial and temporal dynamics of the endothelium. AB - The endothelium is a highly metabolically active organ that is involved in many physiological processes, including the control of vasomotor tone, barrier function, leukocyte adhesion and trafficking, inflammation, and hemostasis. Endothelial cell phenotypes are differentially regulated in space and time. Endothelial cell heterogeneity has important implications for developing strategies in basic research, diagnostics and therapeutics. The goals of this review are to: (i) consider mechanisms of endothelial cell heterogeneity; (ii) discuss the bench-to-bedside gap in endothelial biomedicine; (iii) revisit definitions for endothelial cell activation and dysfunction; and (iv) propose new goals in diagnosis and therapy. Finally, these themes will be applied to an understanding of vascular bed-specific hemostasis. PMID- 15892867 TI - Regulation of smooth muscle differentiation by the myocardin family of serum response factor co-factors. PMID- 15892868 TI - Consolidating the set of known human protein-protein interactions in preparation for large-scale mapping of the human interactome. AB - BACKGROUND: Extensive protein interaction maps are being constructed for yeast, worm, and fly to ask how the proteins organize into pathways and systems, but no such genome-wide interaction map yet exists for the set of human proteins. To prepare for studies in humans, we wished to establish tests for the accuracy of future interaction assays and to consolidate the known interactions among human proteins. RESULTS: We established two tests of the accuracy of human protein interaction datasets and measured the relative accuracy of the available data. We then developed and applied natural language processing and literature-mining algorithms to recover from Medline abstracts 6,580 interactions among 3,737 human proteins. A three-part algorithm was used: first, human protein names were identified in Medline abstracts using a discriminator based on conditional random fields, then interactions were identified by the co-occurrence of protein names across the set of Medline abstracts, filtering the interactions with a Bayesian classifier to enrich for legitimate physical interactions. These mined interactions were combined with existing interaction data to obtain a network of 31,609 interactions among 7,748 human proteins, accurate to the same degree as the existing datasets. CONCLUSION: These interactions and the accuracy benchmarks will aid interpretation of current functional genomics data and provide a basis for determining the quality of future large-scale human protein interaction assays. Projecting from the approximately 15 interactions per protein in the best sampled interaction set to the estimated 25,000 human genes implies more than 375,000 interactions in the complete human protein interaction network. This set therefore represents no more than 10% of the complete network. PMID- 15892869 TI - Genome-scale evidence of the nematode-arthropod clade. AB - BACKGROUND: The issue of whether coelomates form a single clade, the Coelomata, or whether all animals that moult an exoskeleton (such as the coelomate arthropods and the pseudocoelomate nematodes) form a distinct clade, the Ecdysozoa, is the most puzzling issue in animal systematics and a major open ended subject in evolutionary biology. Previous single-gene and genome-scale analyses designed to resolve the issue have produced contradictory results. Here we present the first genome-scale phylogenetic evidence that strongly supports the Ecdysozoa hypothesis. RESULTS: Through the most extensive phylogenetic analysis carried out to date, the complete genomes of 11 eukaryotic species have been analyzed in order to find homologous sequences derived from 18 human chromosomes. Phylogenetic analysis of datasets showing an increased adjustment to equal evolutionary rates between nematode and arthropod sequences produced a gradual change from support for Coelomata to support for Ecdysozoa. Transition between topologies occurred when fast-evolving sequences of Caenorhabditis elegans were removed. When chordate, nematode and arthropod sequences were constrained to fit equal evolutionary rates, the Ecdysozoa topology was statistically accepted whereas Coelomata was rejected. CONCLUSIONS: The reliability of a monophyletic group clustering arthropods and nematodes was unequivocally accepted in datasets where traces of the long-branch attraction effect were removed. This is the first phylogenomic evidence to strongly support the 'moulting clade' hypothesis. PMID- 15892870 TI - Nanoarchaea: representatives of a novel archaeal phylum or a fast-evolving euryarchaeal lineage related to Thermococcales? AB - BACKGROUND: Cultivable archaeal species are assigned to two phyla -- the Crenarchaeota and the Euryarchaeota -- by a number of important genetic differences, and this ancient split is strongly supported by phylogenetic analysis. The recently described hyperthermophile Nanoarchaeum equitans, harboring the smallest cellular genome ever sequenced (480 kb), has been suggested as the representative of a new phylum -- the Nanoarchaeota -- that would have diverged before the Crenarchaeota/Euryarchaeota split. Confirming the phylogenetic position of N. equitans is thus crucial for deciphering the history of the archaeal domain. RESULTS: We tested the placement of N. equitans in the archaeal phylogeny using a large dataset of concatenated ribosomal proteins from 25 archaeal genomes. We indicate that the placement of N. equitans in archaeal phylogenies on the basis of ribosomal protein concatenation may be strongly biased by the coupled effect of its above-average evolutionary rate and lateral gene transfers. Indeed, we show that different subsets of ribosomal proteins harbor a conflicting phylogenetic signal for the placement of N. equitans. A BLASTP-based survey of the phylogenetic pattern of all open reading frames (ORFs) in the genome of N. equitans revealed a surprisingly high fraction of close hits with Euryarchaeota, notably Thermococcales. Strikingly, a specific affinity of N. equitans and Thermococcales was strongly supported by phylogenies based on a subset of ribosomal proteins, and on a number of unrelated molecular markers. CONCLUSION: We suggest that N. equitans may more probably be the representative of a fast-evolving euryarchaeal lineage (possibly related to Thermococcales) than the representative of a novel and early diverging archaeal phylum. PMID- 15892871 TI - Dissection of a DNA-damage-induced transcriptional network using a combination of microarrays, RNA interference and computational promoter analysis. AB - BACKGROUND: Gene-expression microarrays and RNA interferences (RNAi) are among the most prominent techniques in functional genomics. The combination of the two holds promise for systematic, large-scale dissection of transcriptional networks. Recent studies, however, raise the concern that nonspecific responses to small interfering RNAs (siRNAs) might obscure the consequences of silencing the gene of interest, throwing into question the ability of this experimental strategy to achieve precise network dissections. RESULTS: We used microarrays and RNAi to dissect a transcriptional network induced by DNA damage in a human cellular system. We recorded expression profiles with and without exposure of the cells to a radiomimetic drug that induces DNA double-strand breaks (DSBs). Profiles were measured in control cells and in cells knocked-down for the Rel-A subunit of NFkappaB and for p53, two pivotal stress-induced transcription factors, and for the protein kinase ATM, the major transducer of the cellular responses to DSBs. We observed that NFkappaB and p53 mediated most of the damage-induced gene activation; that they controlled the activation of largely disjoint sets of genes; and that ATM was required for the activation of both pathways. Applying computational promoter analysis, we demonstrated that the dissection of the network into ATM/NFkappaB and ATM/p53-mediated arms was highly accurate. CONCLUSIONS: Our results demonstrate that the combined experimental strategy of expression arrays and RNAi is indeed a powerful method for the dissection of complex transcriptional networks, and that computational promoter analysis can provide a strong complementary means for assessing the accuracy of this dissection. PMID- 15892872 TI - The Sequence Ontology: a tool for the unification of genome annotations. AB - The Sequence Ontology (SO) is a structured controlled vocabulary for the parts of a genomic annotation. SO provides a common set of terms and definitions that will facilitate the exchange, analysis and management of genomic data. Because SO treats part-whole relationships rigorously, data described with it can become substrates for automated reasoning, and instances of sequence features described by the SO can be subjected to a group of logical operations termed extensional mereology operators. PMID- 15892873 TI - Synthetic lethal analysis of Caenorhabditis elegans posterior embryonic patterning genes identifies conserved genetic interactions. AB - Phenotypic robustness is evidenced when single-gene mutations do not result in an obvious phenotype. It has been suggested that such phenotypic stability results from 'buffering' activities of homologous genes as well as non-homologous genes acting in parallel pathways. One approach to characterizing mechanisms of phenotypic robustness is to identify genetic interactions, specifically, double mutants where buffering is compromised. To identify interactions among genes implicated in posterior patterning of the Caenorhabditis elegans embryo, we measured synthetic lethality following RNA interference of 22 genes in 15 mutant strains. A pair of homologous T-box transcription factors (tbx-8 and tbx-9) is found to interact in both C. elegans and C. briggsae, indicating that their compensatory function is conserved. Furthermore, a muscle module is defined by transitive interactions between the MyoD homolog hlh-1, another basic helix-loop helix transcription factor, hnd-1, and the MADS-box transcription factor unc-120. Genetic interactions within a homologous set of genes involved in vertebrate myogenesis indicate broad conservation of the muscle module and suggest that other genetic modules identified in C. elegans will be conserved. PMID- 15892874 TI - Relations in biomedical ontologies. AB - To enhance the treatment of relations in biomedical ontologies we advance a methodology for providing consistent and unambiguous formal definitions of the relational expressions used in such ontologies in a way designed to assist developers and users in avoiding errors in coding and annotation. The resulting Relation Ontology can promote interoperability of ontologies and support new types of automated reasoning about the spatial and temporal dimensions of biological and medical phenomena. PMID- 15892875 TI - The Open Microscopy Environment (OME) Data Model and XML file: open tools for informatics and quantitative analysis in biological imaging. AB - The Open Microscopy Environment (OME) defines a data model and a software implementation to serve as an informatics framework for imaging in biological microscopy experiments, including representation of acquisition parameters, annotations and image analysis results. OME is designed to support high-content cell-based screening as well as traditional image analysis applications. The OME Data Model, expressed in Extensible Markup Language (XML) and realized in a traditional database, is both extensible and self-describing, allowing it to meet emerging imaging and analysis needs. PMID- 15892876 TI - A matter of life and death. AB - Our ability to sustain some of the outward signs of life has rendered obsolete the legal system's definition of what it means to be alive. PMID- 15892877 TI - Why genes persist in organelle genomes. AB - Mitochondria and plastids (including chloroplasts) have a small but vital genetic coding capacity, but what are the properties of some genes that dictate that they must remain encoded in organelles? PMID- 15892878 TI - Marsupials and monotremes sort genome treasures from junk. AB - A recent landmark paper demonstrates the unique contribution of marsupials and monotremes to comparative genome analysis, filling an evolutionary gap between the eutherian mammals (including humans) and more distant vertebrate species. PMID- 15892879 TI - Circadian clocks are seeing the systems biology light. AB - Circadian rhythms are those biological rhythms that have a periodicity of around 24 hours. Recently, the generation of a circadian transcriptional network -- compiled from RNA-expression and promoter-element analysis and phase information - has led to a better understanding of the gene-expression patterns that regulate the precise 24-hour clock. PMID- 15892880 TI - The cryptochromes. AB - Cryptochromes are photoreceptors that regulate entrainment by light of the circadian clock in plants and animals. They also act as integral parts of the central circadian oscillator in animal brains and as receptors controlling photomorphogenesis in response to blue or ultraviolet (UV-A) light in plants. Cryptochromes are probably the evolutionary descendents of DNA photolyases, which are light-activated DNA-repair enzymes, and are classified into three groups -- plant cryptochromes, animal cryptochromes, and CRY-DASH proteins. Cryptochromes and photolyases have similar three-dimensional structures, characterized by an alpha/beta domain and a helical domain. The structure also includes a chromophore, flavin adenine dinucleotide (FAD). The FAD-access cavity of the helical domain is the catalytic site of photolyases, and it is predicted also to be important in the mechanism of cryptochromes. PMID- 15892881 TI - The diversity of bacterial pathogenicity mechanisms. AB - A report on the international conference 'Molecular basis of bacterial pathogenesis', sponsored by the Federation of European Microbiological Societies (FEMS) and the Israel Center for the Study of Emerging Diseases, Ein Gedi, Israel, 23-27 January 2005. PMID- 15892882 TI - Identification of ambiguities in the 1994 chronic fatigue syndrome research case definition and recommendations for resolution. AB - BACKGROUND: A recent article by Reeves et al. on the identification and resolution of ambiguities in the 1994 chronic fatigue syndrome (CFS) research case definition recommended the Checklist Individual Strength, the Chalder Fatigue Scale, and the Krupp Fatigue Severity Scale for evaluating fatigue in CFS studies. To be able to discriminate between various levels of severe fatigue, extreme scoring on the individual items of these questionnaires must not occur too often. METHODS: We derived an expression that allows us to compute a lower bound for the number of items with the maximum item score for a given study from the reported mean scale score, the number of reported subjects, and the properties of the fatigue rating scale. Several CFS studies that used the recommended fatigue rating scales were selected from literature and analyzed to verify whether abundant extreme scoring had occurred. RESULTS: Extreme scoring occurred on a large number of the items for all three recommended fatigue rating scales across several studies. The percentage of items with the maximum score exceeded 40% in several cases. The amount of extreme scoring for a certain scale varied from one study to another, which suggests heterogeneity in the selected subjects across studies. CONCLUSION: Because all three instruments easily reach the extreme ends of their scales on a large number of the individual items, they do not accurately represent the severe fatigue that is characteristic for CFS. This should lead to serious questions about the validity and suitability of the Checklist Individual Strength, the Chalder Fatigue Scale, and the Krupp Fatigue Severity Scale for evaluating fatigue in CFS research. PMID- 15892883 TI - Psychological illness is commonly associated with functional gastrointestinal disorders and is important to consider during patient consultation: a population based study. AB - BACKGROUND: Some individuals with functional gastrointestinal disorders (FGID) suffer long-lasting symptoms without ever consulting their doctors. Our aim was to study co-morbidity and lifestyle differences among consulters and non consulters with persistent FGID and controls in a defined adult population. METHODS: A random sample of the general adult Swedish population was obtained by a postal questionnaire. The Abdominal Symptom Questionnaire (ASQ) was used to measure GI symptomatology and grade of GI symptom severity and the Complaint Score Questionnaire (CSQ) was used to measure general symptoms. Subjects were then grouped for study by their symptomatic profiles. Subjects with long-standing FGID (n = 141) and subjects strictly free from gastrointestinal (GI) symptoms (n = 97) were invited to attend their local health centers for further assessment. RESULTS: Subjects with FGID have a higher risk of psychological illness [OR 8.4, CI95(4.0-17.5)] than somatic illness [OR 2.8, CI95(1.3-5.7)] or ache and fatigue symptoms [OR 4.3, CI95(2.1-8.7)]. Subjects with psychological illness have a higher risk of severe GI symptoms than controls; moreover they have a greater chance of being consulters. Patients with FGID have more severe GI symptoms than non-patients. CONCLUSION: There is a strong relation between extra-intestinal, mental and somatic complaints and FGID in both patients and non-patients. Psychological illness increases the chance of concomitantly having more severe GI symptoms, which also enhance consultation behaviour. PMID- 15892884 TI - No evidence for association between polymorphisms in GRM3 and schizophrenia. AB - BACKGROUND: Three studies have previously reported data that were interpreted by the authors as supportive of association between schizophrenia and polymorphisms in the gene encoding the metabotropic glutamate receptor GRM3. METHODS: In a bid to examine this hypothesis, we examined seven SNPs spanning GRM3 in a UK case control sample (schizophrenic cases n = 674, controls n = 716). These included all SNPs previously reported to be associated, alone or in haplotypes, with schizophrenia in European or European American samples. RESULTS: Our data showed no evidence for association with single markers, or 2, 3, 4 and 5 marker haplotypes, nor did any specific haplotypes show evidence for association according to previously observed patterns. CONCLUSION: Examination of our own data and those of other groups leads us to conclude that at present, GRM3 should not be viewed as a gene for which there is replicated evidence for association with schizophrenia. PMID- 15892885 TI - Differences in gene expression in prostate cancer, normal appearing prostate tissue adjacent to cancer and prostate tissue from cancer free organ donors. AB - BACKGROUND: Typical high throughput microarrays experiments compare gene expression across two specimen classes - an experimental class and baseline (or comparison) class. The choice of specimen classes is a major factor in the differential gene expression patterns revealed by these experiments. In most studies of prostate cancer, histologically malignant tissue is chosen as the experimental class while normal appearing prostate tissue adjacent to the tumor (adjacent normal) is chosen as the baseline against which comparison is made. However, normal appearing prostate tissue from tumor free organ donors represents an alterative source of baseline tissue for differential expression studies. METHODS: To examine the effect of using donor normal tissue as opposed to adjacent normal tissue as a baseline for prostate cancer expression studies, we compared, using oligonucleotide microarrays, the expression profiles of primary prostate cancer (tumor), adjacent normal tissue and normal tissue from tumor free donors. RESULTS: Statistical analysis using Significance Analysis of Microarrays (SAM) demonstrates the presence of unique gene expression profiles for each of these specimen classes. The tumor v donor expression profile was more extensive that the tumor v adjacent normal profile. The differentially expressed gene lists from tumor v donor, tumor v adjacent normal and adjacent normal v donor comparisons were examined to identify regulated genes. When donors were used as the baseline, similar genes are highly regulated in both tumor and adjacent normal tissue. Significantly, both tumor and adjacent normal tissue exhibit significant up regulation of proliferation related genes including transcription factors, signal transducers and growth regulators compared to donor tissue. These genes were not picked up in a direct comparison of tumor and adjacent normal tissues. CONCLUSIONS: The up-regulation of these gene types in both tissue types is an unexpected finding and suggests that normal appearing prostate tissue can undergo genetic changes in response to or in expectation of morphologic cancer. A possible field effect surrounding prostate cancers and the implications of these findings for characterizing gene expression changes in prostate tumors are discussed. PMID- 15892886 TI - The unexpected importance of mosquito oviposition behaviour for malaria: non productive larval habitats can be sources for malaria transmission. AB - BACKGROUND: Mosquitoes commute between blood-meal hosts and water. Thus, heterogeneity in human biting reflects underlying spatial heterogeneity in the distribution and suitability of larval habitat as well as inherent differences in the attractiveness, suitability and distribution of blood-meal hosts. One of the possible strategies of malaria control is to identify local vector species and then attack water bodies that contain their larvae. METHODS: Biting and host seeking, not oviposition, have been the focus of most previous studies of mosquitoes and malaria transmission. This study presents a mathematical model that incorporates mosquito oviposition behaviour. RESULTS: The model demonstrates that oviposition is one potential factor explaining heterogeneous biting and vector distribution in a landscape with a heterogeneous distribution of larval habitat. Adult female mosquitoes tend to aggregate around places where they oviposit, thereby increasing the risk of malaria, regardless of the suitability of the habitat for larval development. Thus, a water body may be unsuitable for adult mosquito emergence, but simultaneously, be a source for human malaria. CONCLUSION: Larval density may be a misleading indicator of a habitat's importance for malaria control. Even if mosquitoes could be lured to oviposit in sprayed larval habitats, this would not necessarily mitigate -- and might aggravate -- the risk of malaria transmission. Forcing mosquitoes to fly away from humans in search of larval habitat may be a more efficient way to reduce the risk of malaria than killing larvae. Thus, draining, fouling, or filling standing water where mosquitoes oviposit can be more effective than applying larvicide. PMID- 15892887 TI - Computing expectation values for RNA motifs using discrete convolutions. AB - BACKGROUND: Computational biologists use Expectation values (E-values) to estimate the number of solutions that can be expected by chance during a database scan. Here we focus on computing Expectation values for RNA motifs defined by single-strand and helix lod-score profiles with variable helix spans. Such E values cannot be computed assuming a normal score distribution and their estimation previously required lengthy simulations. RESULTS: We introduce discrete convolutions as an accurate and fast mean to estimate score distributions of lod-score profiles. This method provides excellent score estimations for all single-strand or helical elements tested and also applies to the combination of elements into larger, complex, motifs. Further, the estimated distributions remain accurate even when pseudocounts are introduced into the lod score profiles. Estimated score distributions are then easily converted into E values. CONCLUSION: A good agreement was observed between computed E-values and simulations for a number of complete RNA motifs. This method is now implemented into the ERPIN software, but it can be applied as well to any search procedure based on ungapped profiles with statistically independent columns. PMID- 15892888 TI - The integrins of the urochordate Ciona intestinalis provide novel insights into the molecular evolution of the vertebrate integrin family. AB - BACKGROUND: Integrins are a functionally significant family of metazoan cell surface adhesion receptors. The receptors are dimers composed of an alpha and a beta chain. Vertebrate genomes encode an expanded set of integrin alpha and beta chains in comparison with protostomes such as drosophila or the nematode worm. The publication of the genome of a basal chordate, Ciona intestinalis, provides a unique opportunity to gain further insight into how and when the expanded integrin supergene family found in vertebrates evolved. RESULTS: The Ciona genome encodes eleven alpha and five beta chain genes that are highly homologous to their vertebrate homologues. Eight of the alpha chains contain an A-domain that lacks the short alpha helical region present in the collagen-binding vertebrate alpha chains. Phylogenetic analyses indicate the eight A-domain containing alpha chains cluster to form an ascidian-specific clade that is related to but, distinct from, the vertebrate A-domain clade. Two Ciona alpha chains cluster in laminin-binding clade and the remaining chain clusters in the clade that binds the RGD tripeptide sequence. Of the five Ciona beta chains, three form an ascidian-specific clade, one clusters in the vertebrate beta1 clade and the remaining Ciona chain is the orthologue of the vertebrate beta4 chain. CONCLUSION: The Ciona repertoire of integrin genes provides new insight into the basic set of these receptors available at the beginning of vertebrate evolution. The ascidian and vertebrate alpha chain A-domain clades originated from a common precursor but radiated separately in each lineage. It would appear that the acquisition of collagen binding capabilities occurred in the chordate lineage after the divergence of ascidians. PMID- 15892889 TI - Hippocampal lesions facilitate instrumental learning with delayed reinforcement but induce impulsive choice in rats. AB - BACKGROUND: Animals must frequently act to influence the world even when the reinforcing outcomes of their actions are delayed. Learning with action-outcome delays is a complex problem, and little is known of the neural mechanisms that bridge such delays. When outcomes are delayed, they may be attributed to (or associated with) the action that caused them, or mistakenly attributed to other stimuli, such as the environmental context. Consequently, animals that are poor at forming context-outcome associations might learn action-outcome associations better with delayed reinforcement than normal animals. The hippocampus contributes to the representation of environmental context, being required for aspects of contextual conditioning. We therefore hypothesized that animals with hippocampal lesions would be better than normal animals at learning to act on the basis of delayed reinforcement. We tested the ability of hippocampal-lesioned rats to learn a free-operant instrumental response using delayed reinforcement, and what is potentially a related ability -- the ability to exhibit self controlled choice, or to sacrifice an immediate, small reward in order to obtain a delayed but larger reward. RESULTS: Rats with sham or excitotoxic hippocampal lesions acquired an instrumental response with different delays (0, 10, or 20 s) between the response and reinforcer delivery. These delays retarded learning in normal rats. Hippocampal-lesioned rats responded slightly less than sham-operated controls in the absence of delays, but they became better at learning (relative to shams) as the delays increased; delays impaired learning less in hippocampal lesioned rats than in shams. In contrast, lesioned rats exhibited impulsive choice, preferring an immediate, small reward to a delayed, larger reward, even though they preferred the large reward when it was not delayed. CONCLUSION: These results support the view that the hippocampus hinders action-outcome learning with delayed outcomes, perhaps because it promotes the formation of context outcome associations instead. However, although lesioned rats were better at learning with delayed reinforcement, they were worse at choosing it, suggesting that self-controlled choice and learning with delayed reinforcement tax different psychological processes. PMID- 15892890 TI - Robust detection of periodic time series measured from biological systems. AB - BACKGROUND: Periodic phenomena are widespread in biology. The problem of finding periodicity in biological time series can be viewed as a multiple hypothesis testing of the spectral content of a given time series. The exact noise characteristics are unknown in many bioinformatics applications. Furthermore, the observed time series can exhibit other non-idealities, such as outliers, short length and distortion from the original wave form. Hence, the computational methods should preferably be robust against such anomalies in the data. RESULTS: We propose a general-purpose robust testing procedure for finding periodic sequences in multiple time series data. The proposed method is based on a robust spectral estimator which is incorporated into the hypothesis testing framework using a so-called g-statistic together with correction for multiple testing. This results in a robust testing procedure which is insensitive to heavy contamination of outliers, missing-values, short time series, nonlinear distortions, and is completely insensitive to any monotone nonlinear distortions. The performance of the methods is evaluated by performing extensive simulations. In addition, we compare the proposed method with another recent statistical signal detection estimator that uses Fisher's test, based on the Gaussian noise assumption. The results demonstrate that the proposed robust method provides remarkably better robustness properties. Moreover, the performance of the proposed method is preferable also in the standard Gaussian case. We validate the performance of the proposed method on real data on which the method performs very favorably. CONCLUSION: As the time series measured from biological systems are usually short and prone to contain different kinds of non-idealities, we are very optimistic about the multitude of possible applications for our proposed robust statistical periodicity detection method. PMID- 15892891 TI - Non-isotropic noise correlation in PET data reconstructed by FBP but not by OSEM demonstrated using auto-correlation function. AB - BACKGROUND: Positron emission tomography (PET) is a powerful imaging technique with the potential of obtaining functional or biochemical information by measuring distribution and kinetics of radiolabelled molecules in a biological system, both in vitro and in vivo. PET images can be used directly or after kinetic modelling to extract quantitative values of a desired physiological, biochemical or pharmacological entity. Because such images are generally noisy, it is essential to understand how noise affects the derived quantitative values. A pre-requisite for this understanding is that the properties of noise such as variance (magnitude) and texture (correlation) are known. METHODS: In this paper we explored the pattern of noise correlation in experimentally generated PET images, with emphasis on the angular dependence of correlation, using the autocorrelation function (ACF). Experimental PET data were acquired in 2D and 3D acquisition mode and reconstructed by analytical filtered back projection (FBP) and iterative ordered subsets expectation maximisation (OSEM) methods. The 3D data was rebinned to a 2D dataset using FOurier REbinning (FORE) followed by 2D reconstruction using either FBP or OSEM. In synthetic images we compared the ACF results with those from covariance matrix. The results were illustrated as 1D profiles and also visualized as 2D ACF images. RESULTS: We found that the autocorrelation images from PET data obtained after FBP were not fully rotationally symmetric or isotropic if the object deviated from a uniform cylindrical radioactivity distribution. In contrast, similar autocorrelation images obtained after OSEM reconstruction were isotropic even when the phantom was not circular. Simulations indicated that the noise autocorrelation is non isotropic in images created by FBP when the level of noise in projections is angularly variable. Comparison between 1D cross profiles on autocorrelation images obtained by FBP reconstruction and covariance matrices produced almost identical results in a simulation study. CONCLUSION: With asymmetric radioactivity distribution in PET, reconstruction using FBP, in contrast to OSEM, generates images in which the noise correlation is non-isotropic when the noise magnitude is angular dependent, such as in objects with asymmetric radioactivity distribution. In this respect, iterative reconstruction is superior since it creates isotropic noise correlations in the images. PMID- 15892892 TI - Characterization of TRZ1, a yeast homolog of the human candidate prostate cancer susceptibility gene ELAC2 encoding tRNase Z. AB - BACKGROUND: In humans, mutation of ELAC2 is associated with an increased risk of prostate cancer. ELAC2 has been shown to have tRNase Z activity and is associated with the gamma-tubulin complex. RESULTS: In this work, we show that the yeast homolog of ELAC2, encoded by TRZ1 (tRNase Z 1), is involved genetically in RNA processing. The temperature sensitivity of a trz1 mutant can be rescued by multiple copies of REX2, which encodes a protein with RNA 3' processing activity, suggesting a role of Trz1p in RNA processing in vivo. Trz1p has two putative nucleotide triphosphate-binding motifs (P-loop) and a conserved histidine motif. The histidine motif and the putative nucleotide binding motif at the C-domain are important for Trz1p function because mutant proteins bearing changes to the critical residues in these motifs are unable to rescue deletion of TRZ1. The growth defect exhibited by trz1 yeast is not complemented by the heterologous ELAC2, suggesting that Trz1p may have additional functions in yeast. CONCLUSION: Our results provide genetic evidence that prostate cancer susceptibility gene ELAC2 may be involved in RNA processing, especially rRNA processing and mitochondrial function. PMID- 15892893 TI - Naturally occurring antisense RNA of histone H2a in mouse cultured cell lines. AB - BACKGROUND: An antisense transcript of histone H2a that has no significant protein-coding region has been cloned from a mouse full-length cDNA library. In the present study, we evaluated this transcript by using RT-PCR and compared the expression patterns of the sense and antisense transcripts by using quantitative RT-PCR (qRT-PCR). RESULTS: This antisense RNA was expressed in three mouse cell lines. We call it ASH2a. ASH2a includes not only the complementary sequence of the transcript of Hist2h2aa2 (a replication-dependent histone H2a gene), but also that of the promoter of Hist2h2aa2. The upstream genomic sequence of the transcription start site of the ASH2a-coding gene (ASH2a) lacks both CCAAT and TATA boxes. This absence suggests that the regulation of ASH2a is different from that of the replication-dependent histone H2a genes. Findings from qRT-PCR indicated that the expression pattern of ASH2a was different from that of Hist2h2aa2. Expression of Hist2h2aa2 peaked at 2 to 4 h during S-phase, but that of ASH2a peaked at 1 h. CONCLUSION: We showed the existence of ASH2a, a histone H2a antisense RNA, in mouse cultured cells. The expression pattern of ASH2a is different from that of the sense RNA. PMID- 15892894 TI - Metabolic effect of telmisartan and losartan in hypertensive patients with metabolic syndrome. AB - BACKGROUND: Metabolic syndrome is a cluster of common cardiovascular risk factors that includes hypertension and insulin resistance. Hypertension and diabetes mellitus are frequent comorbidities and, like metabolic syndrome, increase the risk of cardiovascular events. Telmisartan, an antihypertensive agent with evidence of partial peroxisome proliferator-activated receptor activity-gamma (PPARgamma) activity, may improve insulin sensitivity and lipid profile in patients with metabolic syndrome. METHODS: In a double-blind, parallel-group, randomized study, patients with World Health Organization criteria for metabolic syndrome received once-daily doses of telmisartan (80 mg, n = 20) or losartan (50 mg, n = 20) for 3 months. At baseline and end of treatment, fasting and postprandial plasma glucose, insulin sensitivity, glycosylated haemoglobin (HBA1c) and 24-hour mean systolic and diastolic blood pressures were determined. RESULTS: Telmisartan, but not losartan, significantly (p < 0.05) reduced free plasma glucose, free plasma insulin, homeostasis model assessment of insulin resistance and HbAic. Following treatment, plasma glucose and insulin were reduced during the oral glucose tolerance test by telmisartan, but not by losartan. Telmisartan also significantly reduced 24-hour mean systolic blood pressure (p < 0.05) and diastolic blood pressure (p < 0.05) compared with losartan. CONCLUSION: As well as providing superior 24-hour blood pressure control, telmisartan, unlike losartan, displayed insulin-sensitizing activity, which may be explained by its partial PPARgamma activity. PMID- 15892895 TI - Clinicians' management strategies for patients with dyspepsia: a qualitative approach. AB - BACKGROUND: Symptoms from the upper gastrointestinal tract are frequently encountered in clinical practice and may be of either organic or functional origin. For some of these conditions, according to the literature, certain management strategies can be recommended. For other conditions, the evidence is more ambiguous. The hypothesis that guided our study design was twofold: Management strategies and treatments suggested by different clinicians vary considerably, even when optimal treatment is clear-cut, as documented by evidence in the literature. Clinicians believe that the management strategies of their colleagues are similar to their own. METHODS: Simulated case histories of four patients with symptoms from the upper gastrointestinal tract were presented to 27 Swedish clinicians who were specialists in medical gastroenterology, surgery, and general practice and worked at three hospitals in the southern part of Sweden. The patients' histories contained information on the patient's sex and age and the localisation of the symptoms, but descriptions of subjective symptoms and findings from examinations differed from history to history. Interviews containing open-ended questions were conducted. RESULTS: For the same patient, the management strategies and treatments suggested by the clinicians varied widely, as did the strategies suggested by clinicians in the same speciality. Variation was more pronounced if the case history noted symptoms but no organic findings than if the case history noted unambiguous findings and symptoms. However, even in cases with a consensus in the scientific literature on treatment, the variations in clinicians' opinion on management were pronounced. CONCLUSION: Despite these variations, the clinicians believed that the decisions made by their colleagues would be similar to their own. The overall results of this study indicate that we as researchers must make scientific evidence comprehensible and communicate evidence so that clinicians are able to interpret and implement it in practice. Of particular significance is that scientific evidence leads to an evidence-based care which is effective clinical practice and to the promotion of health from the perspective of the patient, together with cost-effectiveness as a priority. PMID- 15892896 TI - Cytoskeleton reorganization mediates alpha6beta1 integrin-associated actions of laminin on proliferation and survival, but not on steroidogenesis of ovine granulosa cells. AB - BACKGROUND: Laminin (LN) is one of the most abundant extracellular matrix components of the basal lamina and granulosa cell layers of ovarian follicles. Culture of ovine granulosa cells (GC) on LN substratum induces cell spreading, enhances cell survival and proliferation, and promotes luteinization. Previous investigations have shown that these effects are mostly mediated by the alpha6beta1 integrin, but its signalization pathways have not been investigated. This study aimed to assess the importance of the cytoskeleton in the alpha6beta1 integrin-mediated actions of laminin on survival, proliferation and steroidogenesis of ovine GC. METHODS: The relationships between morphology and functions of ovine GC cultured on substrata containing LN or/and RGD peptides were investigated. The effects of (1) cytochalasin D, an actin cytoskeleton disrupting drug, (2) a specific function-blocking antibody raised against alpha6 integrin subunit (anti-alpha6 IgG), and (3) an inhibitor of the ERK1/2 signalization pathway (PD98059) were assessed for GC shape, pyknosis and proliferation rates, oestradiol and progesterone secretions. RESULTS: Cytoskeleton disruption by cytochalasin D induced cell rounding, inhibited proliferation, promoted pyknosis, inhibited progesterone secretion and enhanced oestradiol secretion by GC cultured on LN. When GC were cultured on various substrata containing LN and/or RGD peptides in the presence or absence of anti alpha6 IgG, both the existence of close correlations between the percentage of round cells, and the GC proliferation rate (r = -0.87) and pyknotic rate (r = 0.76) were established, but no relationship was found between cell shape and steroidogenesis. Inhibition of the ERK1/2 signalization pathway by PD98059 had no effect on GC shape, proliferation or pyknotic rates. However, it dramatically reduced progesterone secretion, expression of cytochrome P450 cholesterol side chain cleavage and 3beta-hydroxysteroid deshydrogenase enzymes, and enhanced oestradiol secretion, thereby reproducing all the effects of the anti-alpha6 IgG on steroidogenesis of GC cultured on LN. CONCLUSION: LN may participate in the paracrine control of follicular development through different mechanisms. It could enhance proliferation and survival of GC through its alpha6beta1 integrin mediated actions on cytoskeleton. In contrast, its stimulating action on GC luteinization could be partly mediated by the ERK1/2 pathway, irrespective of cell shape. PMID- 15892897 TI - Suppression of colitis-related mouse colon carcinogenesis by a COX-2 inhibitor and PPAR ligands. AB - BACKGROUND: It is generally assumed that inflammatory bowel disease (IBD)-related carcinogenesis occurs as a result of chronic inflammation. We previously developed a novel colitis-related mouse colon carcinogenesis model initiated with azoxymethane (AOM) and followed by dextran sodium sulfate (DSS). In the present study we investigated whether a cyclooxygenase (COX)-2 inhibitor nimesulide and ligands for peroxisome proliferator-activated receptors (PPARs), troglitazone (a PPARgamma ligand) and bezafibrate (a PPARalpha ligand) inhibit colitis-related colon carcinogenesis using our model to evaluate the efficacy of these drugs in prevention of IBD-related colon carcinogenesis. METHODS: Female CD-1 (ICR) mice were given a single intraperitoneal administration of AOM (10 mg/kg body weight) and followed by one-week oral exposure of 2% (w/v) DSS in drinking water, and then maintained on the basal diets mixed with or without nimesulide (0.04%, w/w), troglitazone (0.05%, w/w), and bezafibrate (0.05%, w/w) for 14 weeks. The inhibitory effects of dietary administration of these compounds were determined by histopathological and immunohistochemical analyses. RESULTS: Feeding with nimesulide and troglitazone significantly inhibited both the incidence and multiplicity of colonic adenocarcinoma induced by AOM/DSS in mice. Bezafibrate feeding significantly reduced the incidence of colonic adenocarcinoma, but did not significantly lower the multiplicity. Feeding with nimesulide and troglitazone decreased the proliferating cell nuclear antigen (PCNA)-labeling index and expression of beta-catenin, COX-2, inducible nitric oxide synthase (iNOS) and nitrotyrosine. The treatments increased the apoptosis index in the colonic adenocarcinoma. Feeding with bezafibrate also affected these parameters except for beta-catenin expression in the colonic malignancy. CONCLUSION: Dietary administration of nimesulide, troglitazone and bezafibrate effectively suppressed the development of colonic epithelial malignancy induced by AOM/DSS in female ICR mice. The results suggest that COX-2 inhibitor and PPAR ligands could serve as an effective agent against colitis-related colon cancer development. PMID- 15892898 TI - Factors moderating neuropsychological outcomes following mild traumatic brain injury: a meta-analysis. AB - There continues to be debate about the long-term neuropsychological impact of mild traumatic brain injury (MTBI). A meta-analysis of the relevant literature was conducted to determine the impact of MTBI across nine cognitive domains. The analysis was based on 39 studies involving 1463 cases of MTBI and 1191 control cases. The overall effect of MTBI on neuropsychological functioning was moderate (d = .54). However, findings were moderated by cognitive domain, time since injury, patient characteristics, and sampling methods. Acute effects (less than 3 months postinjury) of MTBI were greatest for delayed memory and fluency (d = 1.03 and .89, respectively). In unselected or prospective samples, the overall analysis revealed no residual neuropsychological impairment by 3 months postinjury (d = .04). In contrast, clinic-based samples and samples including participants in litigation were associated with greater cognitive sequelae of MTBI (d = .74 and .78, respectively at 3 months or greater). Indeed, litigation was associated with stable or worsening of cognitive functioning over time. The implications and limitations of these findings are discussed. PMID- 15892899 TI - Long-term neuropsychological outcomes following mild traumatic brain injury. AB - Mild traumatic brain injury (MTBI) is common, yet few studies have examined neuropsychological outcomes more than 1 year postinjury. Studies of nonreferred individuals with MTBI or studies with appropriate control groups are lacking, but necessary to draw conclusions regarding natural recovery from MTBI. We examined the long-term neuropsychological outcomes of a self-reported MTBI an average of 8 years postinjury in a nonreferred community-dwelling sample of male veterans. This was a cross-sectional cohort study derived from the Vietnam Experience Study. Three groups matched on premorbid cognitive ability were examined, those who (1) had not been injured in a MVA nor had a head injury (Normal Control; n = 3214), (2) had been injured in a motor vehicle accident (MVA) but did not have a head injury (MVA Control; n = 539), and (3) had a head injury with altered consciousness (MTBI; n = 254). A MANOVA found no group differences on a standard neuropsychological test battery of 15 measures. Across 15 measures, the average neuropsychological effect size of MTBI compared with either control group was .03. Subtle aspects of attention and working memory also were examined by comparing groups on Paced Auditory Serial Addition Test (PASAT) continuation rate and California Verbal Learning Test (CVLT) proactive interference (PI). Compared with normal controls, the MTBI group evidenced attention problems in their lower rate of continuation to completion on the PASAT (odds ratio = 1.32, CI = 1.0 1.73) and in excessive PI (odds ratio = 1.66, CI = 1.11-2.47). Unique to the MTBI group, PASAT continuation problems were associated with left-sided visual imperceptions and excessive PI was associated with impaired tandem gait. These results show that MTBI can have adverse long-term neuropsychological outcomes on subtle aspects of complex attention and working memory. PMID- 15892900 TI - The acquisition of face and person identity information following anterior temporal lobectomy. AB - Thirty unilateral anterior temporal lobectomy (ATL) subjects (15 right and 15 left) and 15 controls were presented a multitrial learning task in which unfamiliar faces were paired with biographical information (occupation, city location, and a person's name). Face recognition hits were similar between groups, but the right ATL group committed more false-positive errors to face foils. Both left and right ATL groups were impaired relative to controls in acquiring biographical information, but the deficit was more pronounced for the left ATL group. Recall levels also varied for the different types of biographical information; occupation was most commonly recalled followed by city name and person name. In addition, city and person name recall was more likely when occupation was also recalled. Overall, recall of biographical information was positively correlated with clinical measures of anterograde episodic memory. Findings are discussed in terms of the role of the temporal lobe and associative learning ability in the successful acquisition of new face semantic (biographical) representations. PMID- 15892901 TI - Arithmetic skills and their cognitive correlates in children with acquired and congenital brain disorder. AB - Arithmetic skills and their cognitive correlates were studied in 24 children with myelomeningocele and shunted hydrocephalus (MM), 27 children with severe traumatic brain injuries (TBI), and 26 children with orthopedic injuries (OI). Their average age was 11.56 years (SD = 2.36). They completed the WRAT-3 Arithmetic subtest and a subtraction task consisting of 20 problems of varying difficulty, as well as measures of working memory, declarative memory, processing speed, planning skills, and visuospatial abilities. The MM group performed more poorly on the WRAT-3 Arithmetic subtest and the subtraction task than the other two groups, which did not differ from each other on either measure. The groups did not differ in the number of math fact errors or visual-spatial errors on the subtraction task, but the MM group made more procedural errors than the OI group. The five cognitive abilities explained substantial variance in performance on both arithmetic tests; processing speed, working memory, declarative memory, and planning accounted for unique variance. Exploratory analyses showed that the cognitive correlates of arithmetic skills varied across groups and ages. Congenital and acquired brain disorders are associated with distinct patterns of arithmetic skills, which are related to specific cognitive abilities. PMID- 15892902 TI - Sex hormonal modulation of hemispheric asymmetries in the attentional blink. AB - The present study examines differences in functional cerebral asymmetries modulated by gonadal steroid hormones during the menstrual cycle in women. Twenty one right-handed women with regular menstrual cycles performed a double-stream rapid serial visual presentation (RSVP) task, with one stream in each visual field, during the low steroid menses and the high steroid midluteal phase. They were required to detect a target item, and then a probe item, each of which could appear in either stream. If the probe item appeared 200 ms after the target, detection of the probe was impaired-a phenomenon known as the "attentional blink." This occurred in both streams in the midluteal phase, but only in the right visual field during menses. Thus low steroid levels appeared to restrict the attentional blink to the left hemisphere, while high levels of estradiol and progesterone in the midluteal phase appeared to reduce functional asymmetries by selectively increasing the attentional blink in the right hemisphere. This effect appears to be mediated by estradiol rather than progesterone, and it is compatible with the assumption of a hormone-related suppression of right hemisphere functions during the midluteal phase. PMID- 15892903 TI - Memory retrieval in schizophrenia: evidence from part-list cuing. AB - Schizophrenia patients are known to exhibit episodic verbal memory deficits. Although their neural origin is debated, they have often been compared to the memory problems found in temporal lobe amnesia or frontal lobe dysfunction. Furthermore, it is unclear to what extent such deficits arise at either memory encoding or retrieval. We addressed the issue of retrieval deficits in schizophrenia in a part-list cuing experiment, testing the effect of the presentation of a subset of previously learned material on the retrieval of the remaining items. The part-list cuing procedure generally impairs retrieval but previous work showed that the detrimental effects are more pronounced in amnesic participants than in healthy people, indicating a retrieval deficit under part list cuing conditions in amnesia. In the present study, schizophrenia patients did not exhibit increased susceptibility to part-list cuing effects and thus showed no increased retrieval inhibition from part-list cuing. Moreover, in part list cuing, schizophrenia patients did not mirror the pattern found in amnesia, demonstrating a dissociation between amnesia and schizophrenia patients with respect to this particular memory effect. Implications for the neural basis of the part-list cuing effect and of memory disturbances in schizophrenia are discussed. PMID- 15892904 TI - An evaluation of persisting cognitive effects after withdrawal from long-term benzodiazepine use. AB - Twenty participants with self-reported long-term benzodiazepine use (mean 108 months) who had previously withdrawn from medication (mean 42 months) were administered a battery of neuropsychological tests. Each long-term user was case matched for age, sex, and education to two control participants who reported never taking benzodiazepines (those with and those without anxiety). The results indicated that long-term benzodiazepine use may lead to impairments in the areas of verbal memory, motor control/performance, and nonverbal memory but not visuospatial skills and attention/concentration. The length of abstinence (> 6 months) indicates that these impairments persist well beyond cessation of benzodiazepine use. However, observed impairments in the area of nonverbal memory were not solely attributable to benzodiazepine use and may be influenced by the elevated anxiety levels present in both the case and the anxious control group. PMID- 15892905 TI - Rey's verbal learning test: normative data for 1855 healthy participants aged 24 81 years and the influence of age, sex, education, and mode of presentation. AB - The Verbal Learning Test (VLT; Rey, 1958) evaluates the declarative memory. Despite its extensive use, it has been difficult to establish normative data because test administration has not been uniform. The purpose of the present study was to gather normative data for the VLT for a large number (N = 1855) of healthy participants aged 24-81 years, using a procedure in which the words to be learned were presented either verbally or visually. The results showed that VLT performance decreased in an age-dependent manner from an early age. The learning capacity of younger versus older adults differed quantitatively rather than qualitatively. Females and higher educated participants outperformed males and lower educated participants over the entire age range tested. Presentation mode affected VLT performance differently: auditory presentation resulted in a better recall on Trial 1 (a short-term or working memory measure), whereas visual presentation yielded a better performance on Trial 3, Trial 4, and Delta (a learning measure). PMID- 15892906 TI - Semantic and phonemic sequence effects in random word generation: a dissociation between Alzheimer's and Huntington's disease patients. AB - Alzheimer's disease (AD) patients perform worse on category than letter fluency tasks, while Huntington's disease (HD) patients show the reverse pattern or comparable impairment on both tasks. We developed a random word generation task to further investigate these deficits. Twenty AD and 16 HD patients and 20 elderly and 16 middle-aged controls guessed which of three pictures (hat, cat, or dog) landed on a die's top face sixty times. Three consecutive response pairings were possible: semantic (cat-dog), phonemic (hat-cat), and neutral (hat-dog). Since healthy individuals avoid repeating meaningful associates ("repetition avoidance"), an increased pairing frequency reflects processing deficits. AD patients produced more semantic and HD patients more phonemic pairings compared to their respective control groups, indicating selective semantic and phonemic processing deficits in AD and HD patients, respectively. PMID- 15892907 TI - State-dependent executive deficits among psychopathic offenders. AB - Three hypotheses for cognitive deficits among psychopaths were tested: Response modulation, left hemisphere activation, and an interaction between the 2. Twenty six psychopathic and 32 nonpsychopathic criminal offenders identified with the Hare Psychopathy Checklist-Revised were randomly assigned to left- and right hemisphere activation groups. An auditory processing task was administered, such that the ability to classify nonverbal auditory signals and the ability to manage subgoals were assessed under left- and right-hemisphere activation conditions. The results showed that psychopaths' information processing in general, including response modulation, was deleteriously affected by left-hemisphere activation, supporting 2 of the 3 hypotheses tested. These results offer an explanation for inconsistent findings of executive deficits among psychopaths. PMID- 15892908 TI - Patterns of alcohol use 1 year after traumatic brain injury: a population-based, epidemiological study. AB - This study delineated patterns of alcohol use 1 year after traumatic brain injury (TBI) in a large, population-based, epidemiological, nonclinical sample, and identified predictors of heavy alcohol use in these individuals. Participants were 1,606 adults identified by review of a South Carolina statewide hospital discharge data set, on the basis of satisfying the Centers for Disease Control case definition of TBI, and were interviewed by telephone 1 year after TBI related discharge. Alcohol use in the month prior to interview was classified according to categories from the Quantity-Frequency-Variability Index; heavy drinking was defined as nearly daily use with > or = 5 drinks at least occasionally, or at least three occasions with > or = 5 drinks. A polychotomous logistic regression with 3 response levels (heavy, moderate, and abstinent/infrequent/light drinking) was used to identify predictors of heavy drinking. Heavy drinking in the month prior to interview was reported by 15.4% of participants, while 14.3% reported moderate drinking and 70.3% reported abstinence or light/infrequent drinking. Risk factors for heavy drinking included male gender, younger age, history of substance abuse prior to TBI, diagnosis of depression since TBI, fair/moderate mental health, and better physical functioning. There was no association between drinking patterns and TBI severity. PMID- 15892909 TI - Neurobehavioral effects of harmful algal bloom (HAB) toxins: a critical review. AB - Human exposure to naturally occurring marine toxins has been associated with a range of neurobehavioral abnormalities. The toxins are produced by harmful algal blooms (HABs) and are typically contracted through seafood consumption. The primary target of many of the HAB toxins is the neurologic system, and the neurobehavioral symptoms associated with the HAB illnesses have influenced public health policy. The HAB-related illnesses most frequently linked to neuropsychological disturbance are Amnesic Shellfish Poisoning, Ciguatera Fish Poisoning, and Possible Estuarine Associated Syndrome, which is associated with exposure to the Pfiesteria piscicida organism. Although the neurophysiologic mechanisms underlying many of the HAB illnesses have been well delineated, the literature examining the neuropsychological impairments is unclear and needs to be defined. This review is intended to introduce an emerging area of study linking HAB illnesses with neuropsychological changes. PMID- 15892913 TI - Caffeine: implications for alertness in athletes. AB - Caffeine is one of the most widely consumed drugs in the world, taken socially and for its alertness- and performance-promoting actions. Extensive reports assert that caffeine increases alertness and cognitive performance levels and, when taken before exercise, demonstrates ergogenic properties. Caffeine ingestion has been associated with increased performance during endurance submaximal, and acute, high-intensity exercise. The exact mechanism of action for the performance effects of caffeine is unknown, although several physiologically and psychologically based theories exist as to how caffeine achieves increased performance capabilities. This paper outlines the known sites of caffeine activity in the body,and discusses these with respect to the effects of caffeine observed during performance assessments. PMID- 15892914 TI - The post-lunch dip in performance. AB - For some performance variables, and some individuals, there is a dip in performance during the midafternoon hours (referred to as the post-lunch dip) that is linked to an increase in sleep propensity at that time of day. The post lunch dip is a real phenomenon that can occur even when the individual has had no lunch and is unaware of the time of day. This dip has its roots in human biology, and may be linked to the size of the 12-hour harmonic in the circadian system. It is certainly exacerbated by a high-carbohydrate lunch, and may be more likely to occur in extreme morning-type individuals. PMID- 15892915 TI - Seasonal rhythms and exercise. AB - Levels of leisure-time physical activity and physical fitness are generally higher in the summer than in the winter months for most people living away from the equator. The notion that an abrupt increase in physical activity in the spring, after a period of relative inactivity, can trigger sudden cardiac events has not been confirmed. There are seasonal variations in the physiological responses to exercise and the occurrence of injuries during participation in sports, but it is not known whether these changes are explained by fluctuations in activity levels and environmental conditions, or by any endogenous circannual rhythms in the human. There are indications of endogenous control for some physiological processes (eg, the metabolic responses to a given intensity of exercise) that seem to mediate more favorable effects of exercise on body composition in the winter. Well-trained athletes show obvious seasonality in their competitive performances, generally in line with adopted annual periodization strategies, although these strategies can be disrupted by external seasonal factors, such as heat stress or the susceptibility to upper respiratory tract infections. Maximal oxygen consumption and other physiological indicators of exercise performance might not mirror seasonal variation in real performances, which suggests that top-class athletes maintain a good level of general physical conditioning throughout the year. PMID- 15892916 TI - Seasonal allergy and seasonal decrements in athletic performance. AB - Allergic diseases are common in all age groups and locations around the world. In the United States, allergic diseases affect 20 to 40 million people annually, including 10% to 30% of adults and close to 40% of children. An estimated 15 million people in the United States have been diagnosed as having asthma, with this number on the rise. Concomitant asthma affects 67% of patients who have allergic rhinitis. As a result of the increase in ventilation during exercise, athletes in particular experience significant symptoms of allergy triggered by exposure to aeroallergens. The allergic response causes nasal and conjunctival congestion, tearing, breathing difficulties, pruritus, fatigue, and mood changes, which affect athletic performance. Systemic symptoms of anaphylaxis from allergy, although rare, can be life threatening. Several decades ago it was inconceivable that an athlete who had asthma could perform competitively, let alone win Olympic gold medals. Today, with proper diagnosis, education, and optimal therapeutic management, the allergic athlete can achieve great strides in all sports endeavors. To avoid seasonal allergic flares and maximize performance, the physician providing care for an athlete who has seasonal allergies must be aware of the climatic patterns of aeroallergen expression, and adjust exercise and pharmacologic regimens accordingly. This article summarizes the effects of allergic disease on exercise and highlights the challenges that seasonal allergy place on athletic performance. Doping considerations grant additional complexity to this issue and underscore the need for a competent, skillful, informed, and ethical approach to treating seasonal allergy in the competitive athlete. PMID- 15892917 TI - The menstrual cycle and sport performance. AB - The female sex steroid hormones estrogen and progesterone have potential effects on exercise capacity and performance through numerous mechanisms, such as substrate metabolism, cardiorespiratory function, thermoregulation, psychologic factors, and injuries. Consequently, hormone level changes may theoretically lead to either improved or decreased performance at various times throughout the menstrual cycle. Numerous methodological issues and a paucity of studies have precluded evidence-based conclusions in almost every area of research in this field. In addition, there appears to be a great degree of inter- and intraindividual variability in these hormonal responses. Using oral contraceptives may be advantageous for female athletes who are negatively affected by their menstrual cycle, as they may provide a stable yet controllable hormonal milieu for training and competition. PMID- 15892920 TI - Sleep-wake and other biological rhythms: functional neuroanatomy. AB - Knowledge from traditional neuroanatomical techniques coupled with more recent advances in functional neuroimaging and molecular genetics have led to a better understanding of how structure relates to function of many biological rhythms. In this article we review the current state of knowledge of the functional neuroanatomy of sleep-wake and circadian rhythms and selected infradian rhythms (eg, menstrual and seasonal). We anticipate that progress in our understanding of the molecular biology of sleep and circadian rhythms will lead to the development of novel therapeutic and prophylactic approaches for improved health and quality of life, and for maintaining performance levels during adverse conditions. PMID- 15892921 TI - Sleep, circadian rhythms, and psychomotor vigilance. AB - Psychomotor vigilance performance is highly relevant to athletic performance. It is influenced by a sleep homeostatic process, which builds up pressure for sleep during wakefulness and dissipates this pressure during sleep, and a circadian rhythm process, which produces a waxing and waning of pressure for wakefulness over a 24 hours of the day. During total sleep deprivation, these two processes cause performance to deteriorate progressively over days, modulated within days by further performance reductions at night and relative improvements during the daytime. As the homeostatic pressure for sleep builds up higher across prolonged wakefulness, the rate of dissipation of that pressure during subsequent sleep is enhanced exponentially, so that even brief periods of sleep provide significant performance recuperation. Nevertheless, sleep restriction practiced on a chronic basis induces cumulative performance deficits of the same order of magnitude as observed during total sleep deprivation. There are also considerable individual differences in the degree of vulnerability to performance impairment from sleep loss, and these differences represent a trait. PMID- 15892922 TI - Sleep extension: getting as much extra sleep as possible. AB - Nearly all people, whether they consider themselves sleep deprived or not, can initially obtain extra sleep. However, as accumulating extra sleep reduces carryover sleep debt, a point is reached where it is no longer possible to obtain extra sleep. If there were a practical method to make a precise measurement of a person's daily sleep requirement, it may be possible to show that most individuals are carrying a very large sleep debt. Several observations and studies demonstrate that almost everyone is sleep deprived and carries some amount of sleep debt. How long such an indebtedness will persist without change if no extra sleep is obtained is not known. PMID- 15892923 TI - Insomnia and sleep disruption: relevance for athletic performance. AB - Insomnia is a common sleep complaint even in young adults and has important daytime consequences. Several subjective and objective tools are recommended to assess the magnitude of the problem and to try to find a cause. Chronic insomnia is often caused by precipitating factors, such as acute stress, work conditions, illness, and travel, and perpetuating factors, such as poor sleep hygiene, anxiety, and medications. Insomnia may have implications in athletic performance resulting from physical and cognitive effects. Several pharmacologic and nonpharmacologic approaches are employed in the management of insomnia that have proven effective for short-term treatment. The pharmacologic approaches include the use of zolpidem and specific GABA agonists, benzodiazepines for specific indications, antidepressants, and melatonin. The nonpharmacologic approaches include stimulus control, sleep restriction, relaxation strategies, and cognitive behavioral therapy. PMID- 15892924 TI - Thermophysiologic aspects of the three-process-model of sleepiness regulation. AB - The following overview reconsiders the three-process model of sleepiness regulation (homeostatic, circadian, and sleep inertia) from a thermophysiologic point of view. Our results gathered over the last decade indicate that the homeostatic aspect of sleepiness regulation (ie, buildup of sleepiness during wakefulness and its decay during sleep) is not related to the thermoregulatory system, whereas the two other processes of sleepiness regulation (ie, circadian and sleep inertia process) are clearly related to thermoregulation in humans. Distal skin temperature of hands and feet seems to be the crucial variable for the association between thermophysiology, sleepiness, and sleep. Increased distal skin temperature before a nocturnal sleep episode is a good predictor for short sleep-onset latency. The disappearance of sleep inertia after sleep or a nap episode shows very similar kinetics as distal vasoconstriction. Furthermore, relaxation-induced sleepiness (eg, after lying down, at lights-off, with thermal biofeedback training) also evokes an increase in distal skin temperatures. The reverse effect occurs at lights-on or a posture change from supine to standing, Therefore, in terms of thermophysiology, sleep inertia can be explained as the reverse of a relaxation process (ie, decrease in distal skin temperatures). Our results reinterpret the so-called "sleep-evoked" reduction of core body temperature as a consequence of relaxation-induced vasodilatation after lights off. Sleep per se has no further thermoregulatory effect. Taken together, a thermophysiologic approach may provide a successful strategy to treat sleep-onset insomnia and alleviate sleep inertia. PMID- 15892925 TI - It's practice, with sleep, that makes perfect: implications of sleep-dependent learning and plasticity for skill performance. AB - Although there is no consensus regarding the functions of sleep, one exciting hypothesis is that sleep contributes importantly to learning and memory. Over the last decade, several studies have provided substantive evidence supporting the role of sleep in memory processing. This article focuses on sleep-dependent learning and brain plasticity in humans, specifically in the development of skill performance that is the foundation of many sports actions. The different forms and stages of human memory are discussed, then evidence of sleep-dependent skill learning and associated sleep-dependent brain plasticity is described. In conclusion, a consideration of the fundamental importance of sleep in real-life skill learning is provided. PMID- 15892926 TI - Sleep and circadian rhythms in children and adolescents: relevance for athletic performance of young people. AB - The amount and timing of sleep play significant roles in forming a solid foundation for competitive performance in young athletes. As children mature into and through adolescence, their need for sleep does not decline substantially, although the opportunity to sleep is limited by lifestyle choices, academic and practice schedules, and compelling changes in the biological processes. The biological changes include a more "permissive" pace for the accumulation of sleep pressure across the day in older adolescents and a longer day length in the more mature. These factors all favor later bedtimes and rising times as children pass into adolescence, and a concomitant delay in the optimal timing for waking activities. Among the important threats to athletic performance are insufficient sleep during training and competition and poor appreciation for the best time of day for competitive activities. The specific consequences of these issues for individual athletes are not clear, though when considering young people as a group, support for adequate sleep is a rational intervention to maximize performance. PMID- 15892927 TI - Sleep apnea and sports performance. AB - Obstructive sleep apnea is a significant medical disorder that is increasingly recognized for its wide-ranging effects on physical and mental health. It can have a profound negative impact on sleep quality, daytime alertness, mood, and cardiovascular health, and effects on metabolic and endocrinologic parameters. There is little known about the prevalence and presentation of apnea in athletes and its potential effect on athletic performance. A high index of suspicion for apnea should exist, particularly in those who have anatomic features and symptoms consistent with the disorder. This article reviews pathophysiology, diagnostic modalities, and treatment interventions. PMID- 15892928 TI - Nonpharmacologic techniques for promoting sleep. AB - Athletes could benefit from simple, self-administered, nonpharmacologic techniques for promoting sleep onset. A wealth of physiologic evidence and limited clinical data support several potential methods that might be conveniently applied at or near bedtime. These include inverted posture, skin warming/core cooling, motor relaxation, sensory withdrawal/masking, breathing techniques, and cognitive relaxation. Each holds promise as a possible element of a comprehensive sleep management program, but all need further investigation to confirm their efficacy or to determine optimal methods of application. PMID- 15892929 TI - Effects of exercise on sleep. AB - Historically, perhaps no daytime behavior has been more closely associated with better sleep than exercise. The assumption that exercise promotes sleep has also been central to various hypotheses about the functions of sleep. Hypotheses that sleep serves an energy conservation function, a body tissue restitution function, or a temperature down-regulation function all have predicted a uniquely potent effect of exercise on sleep because no other stimulus elicits greater depletion of energy stores, tissue breakdown, or elevation of body temperature, respectively. Exercise offers a potentially attractive alternative or adjuvant treatment for insomnia. Sleeping pills have a number of adverse side effects and are not recommended for long-term use, partly on the basis of a significant epidemiologic association of chronic hypnotic use with mortality. Other behavioral/cognitive treatments are more effective for chronic insomnia treatment, but difficult and costly to deliver. By contrast, exercise could be a healthy, safe, inexpensive, and simple means of improving sleep. PMID- 15892930 TI - Jet lag and air travel: implications for performance. AB - This review focuses on travel stress in athletes. An outline of the circadian system is presented, followed by an explanation of how disturbances to its regulation are related to jet lag. There are consequences of jet lag that affect exercise performance and health. Measures to ameliorate adverse effects of jet lag include behavioral or pharmacologic strategies. Coaches and mentors and the athletes they support should be considered when preparing for long-haul flights across multiple meridians. PMID- 15892931 TI - Circadian phase shifting, alerting, and antidepressant effects of bright light treatment. AB - Bright light treatment is the most potent melatonin suppressor and circadian phase shifter and is a safe nonpharmacologic antidepressant for seasonal depression. In addition, bright light treatment may restore performance in conditions of sleep debt and misalignment between peak performance and the athletic event. This article discusses the therapeutic use of bright light treatment, its side effects, and mechanisms of action. PMID- 15892932 TI - Sports chronobiology consultation: from the lab to the arena. AB - This final article, coauthored by a chronobiology consultant, a sports psychologist who applied a chronobiology-based program to an Olympic national team, a clinical neurologist, a performance data analyst, a training-conditioning coach from a major league baseball team who applied chronobiology principles to major league pitchers, and a substance abuse expert, discusses practical aspects of a sports chronobiology consultation, including the goals and current arsenal of available interventions. Short vignettes of actual cases are presented for edification, and references are made to appropriate reviews found elsewhere in this issue. PMID- 15892934 TI - Accident patterns and prevention measures for fatal occupational falls in the construction industry. AB - Contributing factors to 621 occupational fatal falls have been identified with respect to the victim's individual factors, the fall site, company size, and cause of fall. Individual factors included age, gender, experience, and the use of personal protective equipment (PPE). Accident scenarios were derived from accident reports. Significant linkages were found between causes for the falls and accident events. Falls from scaffold staging were associated with a lack of complying scaffolds and bodily action. Falls through existing floor openings were associated with unguarded openings, inappropriate protections, or the removal of protections. Falls from building girders or other structural steel were associated with bodily actions and improper use of PPE. Falls from roof edges were associated with bodily actions and being pulled down by a hoist, object or tool. Falls through roof surfaces were associated with lack of complying scaffolds. Falls from ladders were associated with overexertion and unusual control and the use of unsafe ladders and tools. Falls down stairs or steps were associated with unguarded openings. Falls while jumping to a lower floor and falls through existing roof openings were associated with poor work practices. Primary and secondary prevention measures can be used to prevent falls or to mitigate the consequences of falls and are suggested for each type of accident. Primary prevention measures would include fixed barriers, such as handrails, guardrails, surface opening protections (hole coverings), crawling boards/planks, and strong roofing materials. Secondary protection measures would include travel restraint systems (safety belt), fall arrest systems (safety harness), and fall containment systems (safety nets). PMID- 15892935 TI - Contributing factors in construction accidents. AB - This overview paper draws together findings from previous focus group research and studies of 100 individual construction accidents. Pursuing issues raised by the focus groups, the accident studies collected qualitative information on the circumstances of each incident and the causal influences involved. Site based data collection entailed interviews with accident-involved personnel and their supervisor or manager, inspection of the accident location, and review of appropriate documentation. Relevant issues from the site investigations were then followed up with off-site stakeholders, including designers, manufacturers and suppliers. Levels of involvement of key factors in the accidents were: problems arising from workers or the work team (70% of accidents), workplace issues (49%), shortcomings with equipment (including PPE) (56%), problems with suitability and condition of materials (27%), and deficiencies with risk management (84%). Employing an ergonomics systems approach, a model is proposed, indicating the manner in which originating managerial, design and cultural factors shape the circumstances found in the work place, giving rise to the acts and conditions which, in turn, lead to accidents. It is argued that attention to the originating influences will be necessary for sustained improvement in construction safety to be achieved. PMID- 15892936 TI - Manual handling of highway kerbs--focus group findings. AB - The manual handling of concrete highway kerbs remains commonplace in the construction industry despite obvious risks to operatives. This study was undertaken to find out why kerb installation still includes manual handling, to identify the alternatives that exist and to consider how the organisation of the work affects exposure to risk. Focus groups were held with industry professionals to discuss kerb design, installation and the training of operatives, with site visits and interviews undertaken to place the focus group findings in context. The focus group discussions highlighted manufacturer's "myopia", a lack of installation knowledge of designers and shortfalls in training for installation work. It was concluded that risks to the health of construction workers remain as they are not considered in the design of the product, design of the work or ameliorated by adequate risk assessment. Recommendations from the research are that a pro-active approach to health needs to be adopted by the manufacturers of heavy construction products. Designers of work requiring the use of heavy products need to have more experience of site operations, and education and training in manual handling is desirable at all levels in construction organisations. PMID- 15892937 TI - Identification of ergonomics interventions used to reduce musculoskeletal loading for building installation tasks. AB - Skilled workers in the mechanical and electrical installation (M/EI) building and construction trades experience high rates of disabling work-related musculoskeletal disorders (WMSDs). The M/EI trades involve installing piping; heating, ventilation and air conditioning (HVAC), and electrical systems in residential, commercial, and industrial buildings. In the absence of an ergonomics standard in the United States, some building and construction contractors, including M/EI sector contractors, have implemented various ergonomics interventions on their worksites on a voluntary basis. However, no data were available to determine the type of voluntary control measures being implemented, the task-specific hazards for which control measures needed to be developed or refined, and perceived barriers to improving hazard control. As part of a larger effort to obtain this data, the National Institute for Occupational Safety and Health (NIOSH) organized a stakeholder meeting to gather information regarding ergonomics interventions or "best practices" by M/EI contractors and tradespeople. The attendees included 39 industry representatives, 17 construction ergonomics researchers from government and academia, and four ergonomics consultants with experience in the construction industry. Participants spent more than 50% of time meeting in small trade-specific breakout sessions. According to the participants, tasks common to the three trades included (1) drill holes and shoot fasteners; (2) place and install systems, and (3) lift and carry materials and equipment. Engineering interventions described in the stakeholder meeting included tools, equipment, and engineered building materials; administrative controls largely consisted of training and education programs and modifications of work and management practice. Most participants believed that there were significant limits to the impact individual contractors and tradespeople could have in leading ergonomics improvement in the building and construction industry. PMID- 15892938 TI - A system to test the ground surface conditions of construction sites--for safe and efficient work without physical strain. AB - Ground surface conditions on construction sites have an important influence on the health and safety of workers and their productivity. The development of an expert-based "working conditions evaluation" system is described, intended to assist site managers in recognising unsatisfactory ground conditions and remedying these. The system was evaluated in the period 2002-2003. The evaluation shows that companies recognize poor soil/ground conditions as problematic, but are not aware of the specific physical workload hazards. The developed methods allow assessment of the ground surface quality and selection of appropriate measures for improvement. However, barriers exist at present to wide implementation of the system across the industry. Most significant of these is that responsibility for a site's condition is not clearly located within contracting arrangements, nor is it a topic of serious negotiation. PMID- 15892939 TI - Behavioural change phases of different stakeholders involved in the implementation process of ergonomics measures in bricklaying. AB - The objective of this qualitative study was to assess whether a hypothesised sequential order of behavioural change phases would be fulfilled in different groups of stakeholders involved at the start of a process to implement ergonomic [corrected] measures in bricklaying teams. The measures include trestles, bricklaying scaffolds, mast climbing work platforms [corrected] and cranes. The behavioural change phases were: (1) being aware of measures, (2) understanding measures, (3) wanting measures, (4) intention to buy or hire measures, (5) ability to use measures, (6) using measures (experience), and (7) continuing to use measures. Structured interviews were conducted to examine the change phases in two groups of stakeholders (employers/work planners (n=11) [corrected] and foremen/bricklayers (n=9) [corrected] from nine companies) thought to be relevant in the decision to adopt and use the ergonomic [corrected] measures. The results show that the fulfilled behavioural change phases differ between individual stakeholders, groups of stakeholders, companies and also between ergonomic measures. The hypothesised order of fulfilled consecutive behavioural change phases for individual stakeholders has not been confirmed by this study. The relationship between [corrected] fulfilled and unfulfilled change phases by each stakeholder (group) and actual use of each ergonomic measure requires further study, so as to improve the selection of suitable implementation strategies [corrected] PMID- 15892940 TI - The effect of safety initiatives on safety performance: a longitudinal study. AB - Construction industry is one of the most dangerous industries, not only in the USA, but worldwide. In this longitudinal study we examined the effects of safety initiatives on the safety performance of construction companies. One of the measures commonly used in the USA to track a company's safety performance is the experience modification rate (EMR). The EMR is based on the company's safety records (injury claims) from the past three full years and is used to calculate the workers' compensation insurance premiums. In a longitudinal study, we studied the effects of safety efforts and initiatives on the EMR. The results show that safety initiatives and money spent on safety do improve safety performance, but only over time. PMID- 15892941 TI - A multi-level systems approach for the development of tools, equipment and work processes for the construction industry. AB - Ergonomics is a key issue in the construction industry. Many work tasks and associated equipment and tools are not designed with ergonomics principles in mind. Often, in the development of power tools for construction, any attention to ergonomics is restricted to the human-machine interface and handle design. The need for ergonomics intervention in the development process originates from considerations of safety, health, physical work load, and productivity. It is argued that in each of these respects, the construction industry has lower standards than other industries and therefore has a need and opportunity for improvement. A multi-level ergonomics approach is proposed addressing these issues. The approach defines five levels of ergonomics intervention, from designing individual tools for safety, to designing wider aspects of construction and work flow for optimal productivity. This holistic approach is illustrated using case study examples of the development of power tools and work methods. PMID- 15892942 TI - Vibration induced low back disorders--comparison of the vibration evaluation according to ISO 2631 with a force-related evaluation. AB - Long-term vibration stress can contribute to degenerative changes in the joints of the human body, especially in the lumbar spine. An important factor in the development of these diseases is given by the forces transmitted in the joints. Because the forces can hardly be measured a biomechanical model was developed which simulates the human body in the standing and the sitting posture. The vibration properties of the model were adapted to the transfer function provided in the standards and the literature. With the model the compressive forces at the driving point of the body, in the leg joints, and in two motion segments of the spine were simulated under a vertical pseudo random vibration. Transfer functions between the accelerations of the ground or of the seat and the forces were computed. Furthermore, based on the transfer function between seat acceleration and compressive force in the spinal motion segment L3-L4 weighting factors were derived. By means of these factors characteristic vibration values were computed for 57 realistic vibration spectra measured on 17 machines and vehicles. The consideration of the forces resulted in a stronger weighting of low-frequency vibrations compared to the weighted acceleration as suggested by ISO 2631-1. In order to enable an assessment of the health risk a force-related guidance value was derived which amounts to 0.81 ms(-2) (rms). PMID- 15892943 TI - A survey of tools and methods used by certified professional ergonomists. AB - A survey of Certified Professional Ergonomists (CPEs) was conducted to gather information on the types of basic tools, direct and observational measurement techniques, and software used by practitioners. The motivation for the survey was to better understand what types of tools and methods practitioners use, their opinions of these tools, and to potentially gain an understanding of the constraints or preferences that influence this selection. Reasons for using or not using a selection of tools were also surveyed. Of 578 surveys that were delivered to CPEs and Associate Ergonomics Professionals, 308 were returned for a response rate of 53%. The respondents tended to be biased towards physical ergonomics, as the survey primarily focused on this area of ergonomics. A high percentage of respondents reported using tape measures, video cameras, stopwatches and digital cameras. The most commonly used observational methods were those involving manual materials handling, whereas the most commonly used direct measurement tools were pinch and grip dynamometers and push/pull gauges. The frequency and type of checklists, software, and anthropometric data used are summarized. PMID- 15892944 TI - The effect of knife handle shape on stabbing performance. AB - A quantitative knowledge of stabbing ability is a pre-requisite to establishing protection standards for stab resistant body armour. In order to determine the validity of measurements it is necessary to understand all the mechanisms that determine performance. This paper describes a series of tests that were performed in order to determine the effect of handle size and shape on the forces and impact energy that could be produced during stabbing of an armoured target. It was found that the single largest variable was that of the test participants with all other variables such as handle size and shape having only slight effects on the magnitude of impact energy. The use of a finger guard or hilt was shown to increase the mean energy delivered to the target by approximately 5J compared to a handle having no guard. It was also found that the characteristics of energy delivery were strongly influenced by the position of the grip relative to this guard. This reinforces the conclusions of previous work (Horsfall et al., 1999; Chadwick et al., 1999) on the serial nature of momentum transfer during a stabbing impact. PMID- 15892945 TI - Morning warming-up exercise--effects on musculoskeletal fitness in construction workers. AB - The aim of the present study was to evaluate the effects on muscle stretchability, joint flexibility, muscle strength and endurance in construction workers of a 3-month period of a 10-min morning warming-up exercise (MWU), performed at the building site every working day. Thirty construction workers participated in the program. Seventeen construction workers at other building sites served as controls. Muscle stretchability, joint flexibility, muscle strength and endurance were measured before and after the program. Significant increase of thoracic and lower back mobility, increase of hamstring and thigh muscle stretchability were seen in the MWU group. A significant difference in back muscle endurance was found due to decreased endurance in the controls. Muscular strength was not influenced by the MWU. The results indicate that a short dose of morning warming-up exercise could be beneficial for increasing or maintaining joint and muscle flexibility and muscle endurance for workers exposed to manual material handling and strenuous working positions. PMID- 15892946 TI - A review of protein structure and gene organisation for proteins associated with mineralised tissue and calcium phosphate stabilisation encoded on human chromosome 4. AB - Several proteins associated with mineralised tissue (teeth and bone) or involved in calcium phosphate stabilisation in the body fluids, milk and saliva have been mapped to the q arm of human chromosome 4. These include the dentine/bone proteins dentine sialophosphoprotein (DSPP), dentine matrix protein 1 (DMP1), bone sialoprotein (BSP), matrix extracellular phosphoglycoprotein, osteopontin (OPN), enamelin, ameloblastin, milk caseins, salivary statherin, and proline-rich proteins. The proposed function of those that are multiphosphorylated is: (i) the stabilisation of calcium phosphate in solution (e.g. casein, statherin) preventing spontaneous precipitation and seeded-crystal growth or (ii) promoting biomineralisation (e.g. the phosphophoryn domain of DSPP), where the protein described as a template macromolecule, is proposed to act as a nucleator/promoter of crystal growth. The genes of these proteins have been subjected to conserved chromosomal synteny during mammalian evolution. The multiphosphorylated proteins statherin, caseins, phosphophoryn, BSP and OPN have been characterised as intrinsically disordered. The codon usage patterns for the amino acid serine reveal a bias for AGC and AGT codons within the human genes dspp, dmp1 and bsp, mouse dspp and dmp1 but not significantly for statherin or caseins. This pattern was also observed in the gene encoding hen phosvitin that also contains stretches of multiphosphorylated serines and in the dmp1 gene sequences of mammalian, reptilian and avian classes. In conclusion, these intrinsically disordered multiphosphorylated proteins are the translation products of genes displaying examples of codon usage bias, internal repeats and conserved chromosomal synteny within the mammalian class. PMID- 15892947 TI - Variation in dental and skeletal open bite malocclusion in humans with amelogenesis imperfecta. AB - The amelogenesis imperfectas (AI) are a diverse group of genetic disorders primarily affecting the quality and or quantity of enamel, however, affected individuals often have an open bite malocclusion. Three main AI types are recognized based on the perceived developmental mechanisms involved and the enamel phenotype. The purpose of this investigation was to evaluate the association of the AI enamel defect with craniofacial features characteristic of an open bite malocclusion. The sample consisted of 54 AI affected and 34 unaffected family members from 18 different kindreds. Lateral cephalograms were digitized and measurements evaluated for vertical plane alterations using Z scores. Forty two percent of AI affected individuals and 12% of unaffected family members had dental or skeletal open bite malocclusions. Skeletal open bite malocclusion was variably expressed in AI affected individuals. The enamel phenotype severity did not necessarily correspond with the presence or severity of open bite malocclussion. Open bite malocclusion occurred in individuals with AI caused by mutations in the AMELX and ENAM genes even though these genes are considered to be predominantly or exclusively expressed in teeth. Affected AI individuals with cephalometric values meeting our criteria of skeletal open bite malocclusion were observed in all three major AI types. The pathophysiological relationship between AI associated enamel defects and open bite malocclusion remains unknown. PMID- 15892948 TI - Effects of maternal ethanol intake on immunoexpression of epidermal growth factor in developing rat mandibular molar. AB - OBJECTIVE: A polyclonal antibody was used to investigate the effects of ethanol ingestion before and during pregnancy, in the expression of EGF on dentinogenesis and amelogenesis of rat mandibular first molar. DESIGN: Ethanol was administered to drinking water (treated group) starting at concentrations of 1% and increasing weekly to 5, 10, 15, 20 and 25% (v/v). During week 7, these rats were mated and continued to receive the 25% alcoholic solution, up to delivery. The control group received tap water. On postnatal days 0, 4 and 9, two offspring of each litter were killed, their hemimandibles removed and prepared for paraffin processing and immunohistochemistry. RESULTS: At postnatal day 0 the EGF immunoreactivity of the inner enamel epithelium and presecretory ameloblasts was weak when compared to controls. At postnatal day 4 EGF immunoreactivity of the secretory ameloblasts and odontoblasts was only moderate compared to controls. At postnatal day 9 EGF staining of the ameloblasts was weak when compared to controls. CONCLUSIONS: These results suggest that, maternal alcoholism interferes with EGF expression during initial dentinogenesis and amelogenesis and in the secretion and maturation of the dentin and enamel, therefore, which may cause a reduction of dentin and enamel formation. PMID- 15892949 TI - Histological description of tooth formation in adult Eretmodus cf. cyanostictus (Teleostei, Cichlidae). AB - The Eretmodini, a tribe of closely related cichlids (Teleostei, Cichlidae) originating from Lake Tanganyika, possess oral tooth shapes ranging from conical (in Tanganicodus) over cylindrical (in Spathodus) to spatulate (in Eretmodus). Prior to a study aiming to understand how these distinctly different tooth shapes can be acquired in such closely related taxa, a detailed histological study was required of tooth formation in a representative of the eretmodines. Here, we present a histological description of replacement tooth development in Eretmodus cf. cyanostictus. Using light-microscopic observations on semithin as well as on ground sections, microradiographs and stereo-microscopic observations of both alizarine red S stained and unstained jaws we can conclude that tooth formation in adult E. cf. cyanostictus roughly corresponds with what is known for teleost tooth development in general. Remarkable features include the localization and shape of the epithelial downgrowth, the transient presence of a layer intermediate between inner dental epithelium (IDE) and outer dental epithelium (ODE), the asymmetric shape of the enamel organ, the fact that the pulp cavity recedes in front of the forming enameloid during enameloid formation, and finally, the pattern of matrix mineralisation and maturation, and the presence of pigment in the enameloid. The observation that the enamel organ in adult E. cf. cyanostictus develops asymmetrically is significant for understanding tooth shape variation in the Eretmodini. PMID- 15892950 TI - Inhibitory effect of garlic extract on oral bacteria. AB - Garlic (Allium sativum) has long been known to have antibacterial, antifungal and antiviral properties but there are few data on its effects against oral bacterial species particularly putative periodontal pathogens or their enzymes. Filter sterilised, aqueous extract of garlic was tested for ability to inhibit the growth of a range of oral species and to inhibit the trypsin-like and total protease activity Porphyromonas gingivalis. The garlic extract (57.1% (w/v), containing 220 microg/ml allicin) inhibited the growth and killed most of the organisms tested. In general, the minimal inhibitory and minimum bactericidal concentrations for the Gram-negative strains (garlic MIC range 35.7-1.1 mg/ml; allicin mean MIC 4.1 microg/ml; mean MBC 7.9 microg/ml) were lower than those for the Gram-positive strains tested (garlic MIC range 142.7-35.7 mg/ml; allicin mean MIC 27.5 microg/ml; mean MBC 91.9 microg/ml). Also, of the organisms tested, the putative periodontal pathogens had among the lowest MICs (17.8-1.1 mg/ml garlic) and MBCs (35.7-1.1 mg/ml garlic). Time-kill curves for Streptococcus mutans and P. ginigvalis, showed that killing of the latter started almost immediately, whereas there was a delay before S. mutans was killed. The garlic extract also inhibited the trypsin-like and total protease activity of P. gingivalis by 92.7% and 94.88%, respectively. These data indicate that garlic extract inhibits the growth of oral pathogens and certain proteases and so may have therapeutic value, particularly for periodontitis. PMID- 15892951 TI - Impaired induction of cystatin S gene expression by isoproterenol in the submandibular gland of hypophysectomized rats. AB - Cystatin S, an inhibitor of cysteine proteases, is produced and secreted by acinar cells of the rat submandibular gland. Expression of the cystatin S gene is known to be induced at high levels by the beta-adrenergic agonist isoproterenol. In the present study, we revealed that in the submandibular gland of hypophysectomized adult male rats, the levels of induced cystatin S mRNA 24 h after a single administration of isoproterenol are strikingly lower than those in the gland of normal rats. Administration of one of the pituitary-dependent hormones testosterone, estradiol, dexamethasone and thyroxine, together with isoproterenol resulted in marked enhancement of the isoproterenol-induced cystatin S mRNA expression in hypophysectomized rats, whereas administration of any of these hormones alone had no significant effect. These results suggested the existence of cross-talk between the signaling pathways of steroid hormones and isoproterenol in inducing cystatin S gene expression in the rat submandibular gland. PMID- 15892952 TI - Oxidative stress in alcohol-induced rat parotid sialadenosis. AB - This study evaluated the effect of chronic ethanol consumption on the oxidative status of rat parotid and submandibular glands. To identify the endogenous response to ethanol ingestion, the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were determined. In addition, the antioxidant alpha-tocopherol was supplied to the animals in order to estimate its action in ethanol-associated glandular damage. The thiobarbituric acid reactive substances (TBARS), and the protein carbonyl (PC) content, both markers of cellular oxidative stress on lipid and protein structures, respectively, were recorded. Animals subjected to alcohol ingestion showed a low body growth rate with concomitant enlargement of absolute and relative parotid wet weight, compared with pair-fed calorie-controlled rats. Parotid glands of ethanol-treated animals showed increased SOD and GPx activity, and alpha-tocopherol was able to reduce their activities to the control levels. TBARS and PC were enhanced after chronic ethanol treatment in rat parotids. Supplemental alpha-tocopherol suppressed the oxidative ethanol-induced damage in lipid without affecting induced protein oxidation. Submandibular glands revealed no alterations in the weight, enzymatic and oxidative parameters tested due to ethanol and/or alpha-tocopherol ingestion. These findings indicate the involvement of oxidative stress in parotid gland sialadenosis due to ethanol consumption and the capability of alpha-tocopherol to halt lipid damage, although this low-molecular antioxidant compound leads to neither increased glandular weight nor protein oxidation in ethanol-induced parotid alterations. PMID- 15892953 TI - The frequency and distribution of caries in the mediaeval population of Bijelo Brdo in Croatia (10th-11th century). AB - Reconstruction of the life of ancient peoples can be accomplished by studying their dental remains. The further we go into the past, the greater the importance of dental remains for answering a wide spectre of questions related to the life of a particular community. The dental system is a valuable source of information on the type of food, illnesses, and social stratification within a community. The purpose of this paper was to determine the frequency, distribution, and characteristics of dental caries in the mediaeval population of Bijelo Brdo in Croatia. The analysed sample consisted of the dental remains of 85 individuals with the total of 1064 teeth. The majority (979 or 92.0%) of the teeth belonged to the permanent dentition, and this data set was analysed in this report. The frequency of antemortem tooth loss in the sample was 6.7%, the frequency of caries - 9.5%. The most frequent recorded caries were interproximal (3.9%), followed by occlusal (2.9%), and buccal/lingual (1.3%). This research showed that the frequency and distribution of dental caries in the early mediaeval population from Bijelo Brdo is very similar to that of other European populations of the same socio-economic status during the same historic period. Chronological changes in the localisation of caries in populations that inhabited continental Croatia during the mediaeval period indicate a gradual reduction of interproximal caries and an increase of occlusal, buccal, and lingual lesions. These data suggest a change of diet with softer foods becoming more available in the younger time periods. PMID- 15892954 TI - Nonstructural protein 5A does not contribute to the resistance of hepatitis C virus replication to interferon alpha in cell culture. AB - The hepatitis C virus (HCV) subgenomic replicon system was used to study a possible involvement of nonstructural protein 5A (NS5A) in the mechanisms of HCV resistance to interferon alpha (IFN-alpha). A series of chimeric HCV replicons was constructed. In these replicons, the NS5A gene in the backbone of the Con1 replicon was swapped by corresponding fragments obtained from four IFN-alpha responder and four IFN-alpha nonresponder patients that had been infected with the same HCV AD78 strain. Experiments with transfected Huh7 cells did not reveal significant differences in sensitivity of HCV RNA replication to IFN-alpha in cell clones, bearing chimeric Con1/AD78 replicons with NS5A sequences from IFN responders and nonresponders. Thus, these data provide no evidence that the NS5A protein contributes to the resistance of HCV replication to IFN-alpha. PMID- 15892955 TI - Codon volatility does not reflect selective pressure on the HIV-1 genome. AB - Codon volatility is defined as the proportion of a codon's point-mutation neighbors that encode different amino acids. The cumulative volatility of a gene in relation to its associated genome was recently reported to be an indicator of selection pressure. We used this approach to measure selection on all available full-length HIV-1 subtype B genomes in the Los Alamos HIV Sequence Database, and compared these estimates against those obtained via established likelihood- and distance-based comparative methods. Volatility failed to correlate with the results of any of the comparative methods demonstrating that it is not a reliable indicator of selection pressure. PMID- 15892956 TI - Enveloped particles in the serum of chronic hepatitis C patients. AB - HCV particles were isolated from the plasma of chronically infected patients. The virus was analysed by sucrose density gradient centrifugation. The fractions were tested for viral RNA, core antigen and envelope proteins by using a monoclonal antibody directed against the natural E1E2 complex (D32.10). Two populations of particles containing RNA plus core antigen were separated: the first with a density of 1.06-1.08 g/ml did not contain the envelope proteins; the second with a density between 1.17 and 1.21 g/ml expressed both E1 and E2 glycoproteins. Electron microscopy of the enveloped population after immunoprecipitation with D32.10 showed spherical particles with a rather featureless surface and with a diameter around 40 nm. Immuno-gold staining gave evidence that the E1E2 complex was indeed positioned at the surface of these particles. PMID- 15892957 TI - Tyrosine 66 of Pepper vein banding virus genome-linked protein is uridylylated by RNA-dependent RNA polymerase. AB - Pepper vein banding virus (PVBV), a member of the genus potyvirus, is a single stranded positive-sense RNA virus and it primarily infects plants of the family Solanaceae. Genome organization and gene expression strategy of the potyviruses are similar to the picornaviruses, although they infect widely different hosts and have distinctly different morphologies. The genomic RNA of PVBV has a viral genome-linked protein (VPg) at the 5'-terminus and a poly(A) tail at the 3' terminus. In order to establish the role of VPg in the initiation of replication of the virus, recombinant PVBV NIb and VPg were over-expressed in Escherichia coli and purified under non-denaturing conditions. PVBV NIb was found to be active as polymerase and it could uridylylate the VPg in a template independent manner. N- and C-terminal deletion analysis of VPg revealed that N-terminal 21 and C-terminal 92 residues of PVBV VPg are dispensable for in vitro uridylylation. The amino acid residue uridylylated by PVBV NIb was identified to be Tyr 66 by site-directed mutagenesis. It is possible that in potyviruses, replication begins with uridylylation of VPg which acts as primer for progeny RNA synthesis. PMID- 15892958 TI - Lineage extinction and replacement in dengue type 1 virus populations are due to stochastic events rather than to natural selection. AB - Between 1996 and 1998, two clades (B and C; genotype I) of dengue virus type 1 (DENV-1) appeared in Myanmar (Burma) that were new to that location. Between 1998 and 2000, a third clade (A; genotype III) of DENV-1, which had been circulating at that locality for at least 25 years, became extinct. These changes preceded the largest outbreak of dengue recorded in Myanmar, in 2001, in which more than 95% of viruses recovered from patients were DENV-1, but where the incidence of severe disease was much less than in previous years. Phylogenetic analyses of viral genomes indicated that the two new clades of DENV-1 did not arise from the, now extinct, clade A viruses nor was the extinction of this clade due to differences in the fitness of the viral populations. Since the extinction occurred during an inter-epidemic period, we suggest that it was due to a stochastic event attributable to the low rate of virus transmission in this interval. PMID- 15892959 TI - Enhanced functional recovery from spinal cord injury following intrathecal or intramuscular administration of poliovirus replicons encoding IL-10. AB - Poliovirus-based vectors (replicons) have been shown to maintain the in vitro tropism of poliovirus for motor neurons of the CNS. To determine if replicons could be effective for delivery of potentially beneficial proteins to the CNS, we have constructed and characterized a replicon encoding IL-10. IL-10 was rapidly produced in tissue culture cells following in vitro infection with replicons encoding IL-10. Intrathecal inoculation of replicons encoding IL-10 into the non injured CNS of mice transgenic for the poliovirus receptor resulted in expression of IL-10 within motor neurons at 24-48 h post-inoculation, which subsided by 72 96 h post-inoculation. Single intrathecal or intramuscular injections of replicons were given following spinal cord trauma. Animals receiving replicons encoding IL-10 demonstrated a greater functional recovery in the first 24 h after injury that was maintained throughout the testing period. Compared to animals given replicons encoding gfp, CNS tissue from animals given replicons encoding IL 10 revealed extensive expression of IL-10 from astrocytes around the CNS lesion during the first week following injury. The expression of IL-10 from astrocytes also correlated with more resting microglia as opposed to the rounded activated microglia seen in animals given replicons encoding gfp. Results of these studies establish that replicons can be used to express biologically active molecules in motor neurons of the CNS and these biologically active molecules can have a direct effect on the CNS or induce a cascade of molecules that can influence the cellular composition and activation state of cells within the CNS. PMID- 15892960 TI - Thymic pathogenicity of an HIV-1 envelope is associated with increased CXCR4 binding efficiency and V5-gp41-dependent activity, but not V1/V2-associated CD4 binding efficiency and viral entry. AB - We previously described a thymus-tropic HIV-1 envelope (R3A Env) from a rapid progressor obtained at the time of transmission. An HIV-1 molecular recombinant with the R3A Env supported extensive replication and pathogenesis in the thymus and did not require Nef. Another Env from the same patient did not display the same thymus-tropic pathogenesis (R3B Env). Here, we show that relative to R3B Env, R3A Env enhances viral entry of T cells, increases fusion-induced cytopathicity, and shows elevated binding efficiency for both CD4 and CXCR4, but not CCR5, in vitro. We created chimeric envelopes to determine the region(s) responsible for each in vitro phenotype and for thymic pathogenesis. Surprisingly, while V1/V2 contributed to enhanced viral entry, CD4 binding efficiency, and cytopathicity in vitro, it made no contribution to thymic pathogenesis. Rather, CXCR4 binding efficiency and V5-gp41-associated activity appear to independently contribute to thymic pathogenesis of the R3A Env. These data highlight the contribution of unique HIV pathogenic factors in the thymic microenvironment and suggest that novel mechanisms may be involved in Env pathogenic activity in vivo. PMID- 15892961 TI - Inhibition of hepatitis C virus core protein expression in immortalized human hepatocytes induces cytochrome c-independent increase in Apaf-1 and caspase-9 activation for cell death. AB - Hepatitis C virus (HCV) core protein has multifunctional activities. We have previously reported that the core protein of HCV immortalizes primary human hepatocytes, which may relate to multistage hepatocarcinogenic events. These immortalized human hepatocytes (IHH) served as a model to study the mechanism of HCV core protein-mediated cell growth regulation. Inhibition of core protein expression in earlier stages after hepatocyte immortalization leads to the induction of apoptosis. Here, we have observed that introduction of antisense core (AS-Core) sequences for inhibition of core protein expression enhanced the expression of E2F1 and p53 levels in early passage IHH. Inhibition of core protein expression also altered the expression level of Bcl-2 family proteins, displaying an increase of the proapoptotic Bax and a decrease in the level of the anti-apoptotic Bcl-xL proteins. These alterations, however, did not result in the release of cytochrome c from the mitochondria. Apaf-1 is frequently deregulated under various pathologic conditions, and examination of AS-Core-expressing apoptotic cells indicated a significant increase in the level of Apaf-1, which coincided with caspase-9 activation. Knockdown of Apaf-1 or the transcriptional regulatory proteins, E2F1 or p53, by small interfering RNA (siRNA) duplexes inhibited the activation of caspase-9 and enhanced cell viability in AS-Core expressing cells. These findings may contribute to the understanding of the pathophysiology of HCV core protein-mediated hepatocyte growth regulation and disease progression. PMID- 15892962 TI - Protection of beagle dogs from mucosal challenge with canine oral papillomavirus by immunization with recombinant adenoviruses expressing codon-optimized early genes. AB - Replication-deficient adenoviral (rAd5) vaccines containing codon-optimized E1, E2, E4, and E7 genes of canine oral papillomavirus (COPV) were tested singly or in combination to determine which vaccines could protect against mucosal challenge with COPV. In three studies, groups of 4-6 beagle dogs were immunized subcutaneously (s.c.) with 10(11) rAd5 at 8-10 weeks and 4-6 weeks prior to challenge with infectious COPV particles at multiple oral mucosal sites. Control dogs were immunized with equivalent doses of rAd5 expressing human papillomavirus (HPV) type 16 L1 (rAd5-HPV-16 L1). In the first study, complete protection from COPV-induced papillomas was achieved by immunization with rAd5 vaccine combinations expressing either E1 + E2 or E1 + E2 + E4 + E7; whereas two of six dogs immunized with rAd5-E4 + rAd5-E7 and six of six rAd5-HPV16-L1-immunized control dogs developed oral papillomas. In two subsequent studies, rAd5-E1 and rAd5-E2 vaccines were tested singly or in combination to assess levels of protective immunity to COPV challenge. Subcutaneous immunization with either one or two doses of rAd5 expressing the COPV E1 and E2 genes could protect > 90% of challenged dogs from wart formation. In contrast, all eight dogs immunized with rAd5-HPV-16 L1 developed papillomas at multiple sites. Protection was accompanied by significant IFN-gamma responses to COPV E1 and E2 peptides. Partial protection was conferred by two immunizations with either rAd5-E1 (6 of 9 protected) or rAd5 E2 (8 of 9 protected). These data indicate that rAd5 expressing papillomavirus E1 and E2 proteins can induce strong protective responses even in outbred populations under practical immunization conditions. PMID- 15892963 TI - Nef enhances c-Cbl phosphorylation in HIV-infected CD4+ T lymphocytes. AB - The multifunctional HIV-1 protein Nef possesses several motifs that interact with signaling molecules in infected T cells. In order to determine whether Nef influences T cell activation, cells were infected with Nef-positive and Nef negative clones of HIV. CD28 expression and changes in tyrosine phosphorylation were monitored. We observed no Nef-dependent changes in CD28 expression or function. However, infection with Nef-positive virus led to changes in tyrosine phosphorylation. This Nef-induced phosphorylation was observed in unstimulated cells, and c-Cbl was identified as one of the proteins whose phosphorylation was upregulated by Nef. Furthermore, Lck is required for Nef-mediated c-Cbl tyrosine phosphorylation. These results suggest that Nef modifies T cell signaling in the absence of T cell receptor engagement and co-stimulation. PMID- 15892964 TI - An arthritogenic alphavirus uses the alpha1beta1 integrin collagen receptor. AB - Ross River (RR) virus is an alphavirus endemic to Australia and New Guinea and is the aetiological agent of epidemic polyarthritis or RR virus disease. Here we provide evidence that RR virus uses the collagen-binding alpha1beta1 integrin as a cellular receptor. Infection could be inhibited by collagen IV and antibodies specific for the beta1 and alpha1 integrin proteins, and fibroblasts from alpha1 integrin-/- mice were less efficiently infected than wild-type fibroblasts. Soluble alpha1beta1 integrin bound immobilized RR virus, and peptides representing the alpha1beta1 integrin binding-site on collagen IV inhibited virus binding to cells. We speculate that two highly conserved regions within the cell receptor binding domain of E2 mimic collagen and provide access to cellular collagen-binding receptors. PMID- 15892965 TI - Poinsettia latent virus is not a cryptic virus, but a natural polerovirus sobemovirus hybrid. AB - The biochemical and genetic features of Poinsettia latent virus (PnLV, formerly named Poinsettia cryptic virus), which is spread worldwide in commercial cultivars of Euphorbia pulcherrima without inducing symptoms, have been determined using virus-purification, immunological techniques, electron microscopy, cloning, and sequencing. PnLV was found to be a chimeric virus with one 4652 bases, plus strand RNA showing a close relationship to poleroviruses within the first three quarters of its genome but to sobemoviruses in the last quarter. Thus, we propose to classify this virus as "polemovirus". Similarities of protein and nucleic acid sequences at the 5' and extreme 3' end of its RNA suggest a replication mode like that of poleroviruses, whereas the coat protein sequence is closely related to that of sobemoviruses. Consistent with these results, PnLV forms stable icosahedra of 34 nm in diameter. The consequences for the taxonomy of PnLV and for gardeners' practice are discussed. PMID- 15892966 TI - Natural resistance of human immunodeficiency virus type 2 to zidovudine. AB - Zidovudine (AZT) is a reverse transcriptase (RT) inhibitor widely used to treat persons infected with HIV-1 and HIV-2. Recent data on treated patients suggest differences in the antiviral activity of AZT between HIV-1 and HIV-2. We evaluated the antiviral activity of AZT on HIV-2 by using multiple approaches including in vitro selection experiments, analysis of growth kinetics with AZT, and phenotypic testing. A total of 5 wild-type (WT) HIV-2 viruses were used in the analysis. For comparison, 4 control WT HIV-1 strains and one HIV-1 mutant carrying the 215S mutation were evaluated in parallel. All 5 HIV-1 isolates acquired AZT resistance mutations after 3-6 passages with AZT or a 4- to 32-fold increase in AZT concentration. Among these viruses, the fastest selection of resistance was seen in HIV-1(S215), which acquired S215Y (1-nucleotide change only) at passage 3 after only 17 days in culture. In contrast, none of the 5 HIV 2 viruses that naturally have S215 acquired S215Y/F or any other RT mutation during 10 passages with AZT (1025-fold increase in AZT concentration). In the presence of AZT + didanosine (ddI), 3 of the 5 HIV-1 isolates acquired AZT or ddI resistance mutations, while only ddI resistance mutations were seen in HIV-2 (4 of 5 isolates). All HIV-2 viruses replicated efficiently in high AZT concentrations and were about 200-fold less sensitive to AZT than HIV-1. In contrast, HIV-2 and HIV-1 were equally susceptible to ddI, a finding consistent with the selection of HIV-2 mutants with AZT + ddI. Our results demonstrate that the activity of AZT on HIV-2 is lower than previously thought, and emphasize the need for novel antiretroviral drugs specific for HIV-2. PMID- 15892967 TI - Adeno-associated virus type 2 enhances goose parvovirus replication in embryonated goose eggs. AB - The autonomous goose parvovirus (GPV) and the human helper-dependent adeno associated virus type 2 (AAV2) share a high degree of homology. To determine if this evolutionary relationship has a biological impact, we studied viral replication in human 293 cells and in embryonated goose eggs coinfected with both viruses. Similar experiments were performed with the minute virus of mice (MVM), an autonomous murine parvovirus with less homology to AAV2. In human 293 cells, both GPV and MVM augmented AAV2 replication. In contrast, AAV2 markedly enhanced GPV replication in embryonated goose eggs under conditions where a similar effect was not observed with MVM. AAV2 did not replicate in embryonated goose eggs and AAV2 inactivated by UV-irradiation also enhanced GPV replication. To our knowledge, this is the first report that a human helper-dependent member of the Parvoviridae can provide helper activity for an autonomous parvovirus in a natural host. PMID- 15892968 TI - African green monkey kidney Vero cells require de novo protein synthesis for efficient herpes simplex virus 1-dependent apoptosis. AB - During HSV-1 infection, IE gene expression triggers apoptosis, but subsequent synthesis of infected cell proteins blocks apoptotic death from ensuing. This "HSV-1-dependent" apoptosis was identified in HEp-2/HeLa cells infected with wild type HSV-1 in the presence of an inhibitor of protein synthesis or a virus lacking ICP27 {HSV-1(vBSDelta27)}. Unlike HEp-2/HeLa cells, vBSDelta27-infected Vero cells fail to exhibit dramatic apoptotic morphologies at times prior to 24 hpi. Here, we examined the basis of these different apoptotic responses to HSV-1. We found that infected Vero cells take substantially longer than HEp-2/HeLa cells to display membrane blebbing, chromatin condensation, DNA laddering, and PARP cleavage. Vero, but not HEp-2/HeLa, cells required de novo protein synthesis to exhibit efficient HSV-1-dependent apoptosis, which included changes in mitochondrial membrane potential, and these factors were produced prior to 3 hpi. Vero cells infected with recombinant viruses devoid of the ICP27 and ICP4 proteins alone or both the ICP27 and ICP22 proteins were apoptotic. These results indicate a requirement for cellular or other viral protein synthesis in Vero cells and provide insight into cell type differences in HSV-1-dependent apoptosis. PMID- 15892969 TI - Functional characterization of Ebola virus L-domains using VSV recombinants. AB - VSV recombinants containing the overlapping L-domain sequences from Ebola virus VP40 (PTAPPEY) were recovered by reverse-genetics. Replication kinetics of M40 WT, M40-P24L, and M40-Y30A were indistinguishable from VSV-WT in BHK-21 cells, whereas the double mutant (M40-P2728A) was defective in budding. Insertion of the Ebola L-domain region into VSV M protein was sufficient to alter the dependence on host proteins for efficient budding. Indeed, M40 recombinants containing a functional PTAP motif specifically incorporated endogenous tsg101 into budding virions and were dependent on tsg101 expression for efficient budding. Thus, VSV represents an excellent negative-sense RNA virus model for elucidating the functional aspects and diverse host interactions associated with the L-domains of Ebola virus. PMID- 15892970 TI - The RNA helicase DDX1 is involved in restricted HIV-1 Rev function in human astrocytes. AB - Productive infection by human immunodeficiency virus type I (HIV-1) in the central nervous system (CNS) involves mainly macrophages and microglial cells. A frequency of less than 10% of human astrocytes is estimated to be infectable with HIV-1. Nonetheless, this relatively low percentage of infected astrocytes, but associated with a large total number of astrocytic cells in the CNS, makes human astrocytes a critical part in the analyses of potential HIV-1 reservoirs in vivo. Investigations in astrocytic cell lines and primary human fetal astrocytes revealed that limited HIV-1 replication in these cells resulted from low-level viral entry, transcription, viral protein processing, and virion maturation. Of note, a low ratio of unspliced versus spliced HIV-1-specific RNA was also investigated, as Rev appeared to act aberrantly in astrocytes, via loss of nuclear and/or nucleolar localization and diminished Rev-mediated function. Host cellular machinery enabling Rev function has become critical for elucidation of diminished Rev activity, especially for those factors leading to RNA metabolism. We have recently identified a DEAD-box protein, DDX1, as a Rev cellular co-factor and now have explored its potential importance in astrocytes. Cells were infected with HIV-1 pseudotyped with envelope glycoproteins of amphotropic murine leukemia viruses (MLV). Semi-quantitative reverse transcriptase-polymerase chain reactions (RT-PCR) for unspliced, singly-spliced, and multiply-spliced RNA clearly showed a lower ratio of unspliced/singly-spliced over multiply-spliced HIV-1-specific RNA in human astrocytes as compared to Rev-permissive, non-glial control cells. As well, the cellular localization of Rev in astrocytes was cytoplasmically dominant as compared to that of Rev-permissive, non-glial controls. This endogenous level of DDX1 expression in astrocytes was demonstrated directly to lead to a shift of Rev sub-cellular distribution dominance from nuclear and/or nucleolar to cytoplasmic, as input of exogenous DDX1 significantly altered both Rev sub cellular localization from cytoplasmic to nuclear predominance and concomitantly increased HIV-1 viral production in these human astrocytes. We conclude that altered DDX1 expression in human astrocytes is, at least in part, responsible for the unfavorable cellular microenvironment for Rev function in these CNS-based cells. Thus, these data suggest a molecular mechanism(s) for restricted replication in astrocytes as a potential low-level site of residual HIV-1 in vivo. PMID- 15892971 TI - Stimulation of cell motility by a viral fibroblast growth factor homolog: proposal for a role in viral pathogenesis. AB - The Autographa californica M nucleopolyhedrovirus (AcMNPV) encodes a gene (open reading frame 32) with homology to vertebrate and invertebrate fibroblast growth factors (fgfs), key regulators of developmental processes affecting the growth, differentiation, and migration of many cell types. We studied the temporal regulation of the AcMNPV fgf, vfgf, by Northern (RNA) blot hybridization; vfgf was transcribed as a 0.6-kb mRNA at early times but as part of a 1.4-kb bicistronic mRNA at late times. The product of vfgf, vFGF, exhibited a number of characteristics that have also been demonstrated for other FGF homologs. vFGF had strong affinity to heparin, a property important for FGF signaling via an FGF receptor. vFGF was secreted into the extracellular fluid when expressed in insect cells, suggesting that it acts as an extracellular ligand. Finally, vFGF was able to stimulate migration of several different types of insect cells. We discuss how this activity may be important for its function during virus infection. PMID- 15892972 TI - A single amino acid change in the C-terminal domain of the matrix protein M1 of influenza B virus confers mouse adaptation and virulence. AB - Serial passage of an initially avirulent influenza B virus, B/Memphis/12/97, resulted in the selection of a variant which was lethal in mice. Virulence correlated with improved growth in vivo and prolonged replication. Sequencing of the complete coding regions of the parent and mouse-adapted viruses revealed 8 amino acid differences. Sequencing and characterization of intermediate passages suggested that one change in the C-terminal domain of the M1 protein, an asparagine to a serine at position 221, was responsible for acquisition of virulence and lethality. Site-directed mutagenesis of the M segment of a different virus, B/Yamanashi/166/98, to change this amino acid residue confirmed its importance by conferring improved growth and virulence in mice. This observation suggests a role for the C domain of the M1 protein in growth and virulence in a mammalian host. PMID- 15892973 TI - New technology for the detection of pH. AB - The measurement of pH is the most widely employed test for (bio-)chemical lab. Since the first use of glass electrode to detect pH, new techniques and methods have broaden the scope of pH detection. Metal/metal oxide, ion sensitive field effect transistors (ISFET), fibre-optical techniques, nanotechniques, and conducting polymer techniques have been extensively developed. This review covers the various methods for pH detection. New development trends were discussed. PMID- 15892974 TI - A method of spectral subband decomposition by simultaneous fitting the initial spectrum and a set of its derivatives. AB - An improved method for spectral subband decomposition based on simultaneous fitting of the initial spectrum and a set of its derivatives is introduced. Additionally, a procedure for finding an optimal smoothing filter to obtain undistorted derivatives is suggested. The proposed method is demonstrated with a model spectrum as well as with experimental absorption spectra of the photosynthetic antenna complexes, peridinin-chlorophyll a-protein (PCP) and the main light-harvesting complex of higher plants (LHC II). PMID- 15892975 TI - Rapid screening procedures for identification of succinic acid producers. AB - Succinic acid, an intermediate of tricarboxylic acid cycle, is produced and accumulated by anaerobic microorganisms. The long-standing interest in the production of this organic acid is because it is a key compound in producing more than 30 commercially important products. The detection of succinic acid is generally carried out by gas chromatography (GC), enzymatic assays, ion-exclusion chromatography (IEC) or by high performance liquid chromatography (HPLC). However, these methods are time consuming, require sophisticated instrumentation and are expensive. In the present investigation we are reporting two rapid, cost effective screening methods for the detection of this important organic acid. These methods can be utilized to screen a large number of microbes producing succinic acid in a very short span of time. PMID- 15892976 TI - Synthesis and characterization of a novel reagent containing dansyl group, which specifically alkylates sulfhydryl group: an example of application for protein chemistry. AB - We have synthesized a novel reagent containing dansyl group, iodoacethyl dansylcadaverine (IADC), which specifically alkylates sulfhydryl groups. The carboxyl group of iodoacetic acid was activated with dicyclohexylcarbodiimide and was condensed with amino group of dansylcadaverine. Purity and chemical structure of IADC was confirmed with mass spectrometry (MS) and NMR. IADC alkylated GSH but not GSSG, which was confirmed by MS. The reactivity of IADC with proteins was also investigated with Western blotting using anti-dansyl antibody. IADC reacted only with sulfhydryl-containing proteins. The specificity of the interaction of IADC with sulfhydryl groups in proteins was confirmed by adding excessive amount of a well-known sulfhydryl-specific reagent, 5, 5'-dithiobis(2-nitrobenzoic acid), which led to a complete inhibition. To show the usefulness of IADC, the cysteines in glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from chicken muscle were modified with this reagent, and GAPDH was then digested by lysyl endopeptidase. The peptides generated from digestion of IADC-incorporated GAPDH were applied to an anti-dansyl immunoaffinity column. The peptide fragments bound and eluted from the column were separated by HPLC, and the amino acid sequence of each peptide was analyzed, and peptide was identified as the one containing a Cys residue(s). These data showed that IADC is a useful reagent to specifically identify the positions of a Cys residue(s) in proteins. PMID- 15892977 TI - Antiserum specific for the intact isoform-3 of metallothionein. AB - The recombinant form of isoform-3 of mouse brain metallothionein (MT3) was used as an antigen to immunize rabbits and raise MT3-selective antiserum. The antiserum was essentially specific for MT3 with 100-fold greater sensitivity for MT3 compared to MT1 or MT2. Immunonblot analysis of whole mouse brain homogenates showed that MT3 was present only in the fraction retained by a 30,000-Da cut-off filter. The antiserum was used to immunoprecipitate MT3 from mouse brain extracts of Swiss Webster mice and provided evidence that MT3 was a member of a macromolecular complex of greater than 30,000 Da mass in brain. An ELISA was developed using purified, recombinant mouse brain Cd(7)-MT3 as the antigen and used to quantify MT3 in mouse brain extracts. The concentration of MT3 was found to be 3.0+/-0.8 microg/ml or approximately 3.5 microg/g mouse brain (wet weight). PMID- 15892978 TI - Single cell electrophoresis in determining cell death: potential for use in organ transplant research. AB - Viability of donor tissues is essential for the success of organ transplantation. Although much work has been done in the field of organ preservation, currently there are few objective methods for evaluating transplant organ viability, and thus preservation efficiency. In the field of cancer biology, single-cell gel electrophoresis (SCGE) is a technique commonly used to measure the efficacy of anti-tumor treatments by measuring the breakdown of tumor cell deoxyribonucleic acid (DNA). This assay has recently been applied to various organs from a postmortem porcine animal model, and cells were found to undergo postmortem breakdown in a way similar to apoptosis-induced DNA fragmentation. Collections of cells from each organ reached levels indicative of non-viability as the postmortem interval (PMI) progressed. The rates of cellular DNA degradation were found to be specific to each organ type at a given ambient temperature. We believe that following the application of various preservation techniques, SCGE assay has the potential to provide a clear indication of cell viability in an organ destined for transplant. As a readily available viability assay, this technique could provide transplant researchers with a useful tool to quantify the efficacy of their experimental organ preservation techniques. PMID- 15892979 TI - Quantitating adenylate nucleotides in diverse organisms. AB - Quantitation of cellular adenylate levels (i.e., ATP, ADP, AMP) has widespread applications in physiological, metabolic and energetic studies. We have compared classical adenylate extraction procedures (i.e., perchloric acid, boiling) with a previously unreported proteinase K-based extraction technique. Our results suggest that all three techniques are comparable in soft animal tissue, but proteinase K-based extractions consistently generated higher adenylate yields from a broad range of organisms, particularly those containing a cell wall (e.g., alga, bacteria, fungi, plant). PMID- 15892980 TI - Stress-induced cortisol elevations are associated with impaired delayed, but not immediate recall. AB - Glucocorticoids are known to modulate memory functions, with elevated cortisol levels being associated with impaired declarative memory. This specific effect has been shown in several studies using pharmacological doses of cortisol. The present study was designed to assess the effects of stress-induced cortisol elevations on (1) the type of memory processing (encoding, consolidation and retrieval), and (2) on the emotional valence of the material under study. Sixteen healthy females were presented neutral and emotional material (words and paragraphs) before and after a stress challenge. Declarative memory was tested immediately after presentation and 24 h later (delayed recall). Delayed, but not immediate recall of the information presented after the stress challenge was significantly reduced compared with delayed recall of information presented before the stress challenge. In line with this, strong negative correlations were found for delayed recall of words and spatial memory presented after the challenge with post-stress cortisol levels, whereas no significant correlations were found between cortisol levels and delayed recall at day 1. These results suggest that stress-induced cortisol specifically affects long-term consolidation of declarative memories. These findings may have implications for understanding the effects of traumatic stress on memory functioning in patients with stress related psychiatric disorders. PMID- 15892981 TI - Reduced word-repetition effect in the event-related potentials of thought disordered patients with schizophrenia. AB - Repetition effect in event-related potentials (ERPs) was studied in 10 non thought-disordered (non-TD) patients with schizophrenia, 8 thought-disordered (TD) patients with schizophrenia, and 10 normal control subjects while they performed a semantic categorization task with incidental word repetitions. All patients were in a stable or partially remitted stage. Although both healthy control and non-TD groups produced more positive ERPs to the repeated words than to the new words (ERP repetition effect) for 250-500 ms, the TD group did not show the ERP repetition effect. These findings suggest that the abnormal attenuation of the ERP repetition effect during semantic processing may be more prominent in schizophrenic patients with thought disorder than in those without the symptom. PMID- 15892982 TI - Verbal fluency in institutionalized patients with schizophrenia: age-related performance decline. AB - Several studies have reported a relatively stable level of cognitive deficits among patients with schizophrenia regardless of age, while others have suggested continued deterioration with age. We compared the performance of 42 institutionalized patients with schizophrenia and 42 age- and education-matched healthy controls on a semantic and phonemic verbal fluency test. Each group was divided into young participants (<65 years old) and elderly participants (> or =65 years old). We found a fluency condition x diagnostic group x age group interaction on total words produced, a fluency condition x diagnostic group interaction on the number of cluster-related words, and a fluency condition x age group interaction on the number of switches. Patients with schizophrenia generally used similar strategies (i.e., semantic or phonemic cluster-related words and switches) as healthy individuals when generating words, but to a lesser degree. We found a disproportionate decline in the elderly schizophrenic patients relative to that of healthy controls only on the phonemic, relative to the semantic test. This decline in performance appears related to the effects of aging rather than severity or chronicity of illness, duration of institutionalization, or a progressive degenerative process associated with the disorder. PMID- 15892983 TI - Comparisons between psychosis samples with different patterns of substance use recruited for clinical and epidemiological studies. AB - Despite high rates of comorbid substance use disorders and smoking among people with psychotic disorders, previous studies have not examined differences in socio demographic, clinical or disability characteristics between psychosis sub-groups with different patterns of substance use. This study compared the characteristics of five groups of people with psychosis and varying patterns of substance use (n=1152), including groups entering treatment studies for substance use or smoking, epidemiological samples of substance users and smokers, and people without such problems. Data were drawn from several recent Australian studies using comparable structured interviews and scales. There were moderate group differences in illness and symptom profiles, with substance users tending to have higher depression and reality distortion scores. Unexpectedly, personal disability was considerably lower among those seeking treatment for their substance use compared with the epidemiological groups, raising concerns about the appropriateness of traditional recruitment approaches in treatment studies and highlighting the need for more assertive treatment engagement and referral strategies in routine clinical settings. As a consequence of uncertainty about links between substance use and everyday functioning, it is suggested that health messages to young people may need to strike a better balance between the potential harms and benefits associated with substance use in pursuing broader harm minimization goals. PMID- 15892984 TI - Effects of serotonin and serotonergic agonists and antagonists on the production of tumor necrosis factor alpha and interleukin-6. AB - Serotonin (5-HT) is a neurotransmitter and immune modulator. The effect of 5-HT on the production of cytokines by human macrophages and lymphocytes is poorly recognized. In the present article we examine the role of 5-HT in modulating the production of two pro-inflammatory cytokines, i.e. interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFalpha), as well as the role of 5-HT(1A) and 5-HT(2) receptors in this process. The specific aims were to examine the effects of 5-HT, p-chlorophenylalanine (PCPA), a 5-HT depleting agent, flesinoxan, a 5-HT(1A) agonist, m-chlorophenylpiperazine (mCPP), a 5-HT(2B/2C) agonist, and ritanserin, a 5-HT(2A/2C) antagonist, on the production of the above cytokines. We found that: (1) 5-HT, 15 microg/ml, significantly decreased IL-6 and TNFalpha production; (2) pCPA, 5 microM, significantly suppressed the production of IL-6 and TNFalpha; and (3) mCPP, 2.7 microg/ml, significantly increased the production of IL-6 and TNFalpha. It is concluded that intracellular 5-HT is necessary for optimal synthesis of IL-6 and TNFalpha; 5-HT in physiological concentrations may increase IL-6 and TNFalpha production by stimulating 5-HT(2) receptors; and extracellular 5-HT concentrations above the baseline physiological levels may suppress the production of the above cytokines. PMID- 15892985 TI - Impulsivity but not venturesomeness is related to morningness. AB - The Composite Scale of Morningness (CSM) is a psychometrically sound instrument available in several languages, including French, aimed at arranging individuals along a continuum from high "eveningness" to high "morningness." On the other hand, impulsivity is involved in many personality disorders and is thought to be orthogonal to venturesomeness, which can be seen as a component of the broader construct of sensation seeking. We hypothesized that evening-type subjects would be more impulsive than morning-types. Self-administered questionnaires were distributed to students, and only complete forms were analyzed (194 males and 358 females). A four-way analysis of covariance showed significant effects of age, gender and impulsivity, but not venturesomeness, on morningness in the sense of a higher degree of eveningness in more impulsive subjects. In addition, the correlation coefficients in both genders were similar to those reported in smaller samples. Our findings deserve further interest because, regardless of gender, they suggest a possible physiopathological approach to impulsivity that may be accessible by circadian interventions such as midday bright light exposure or pharmacological treatments. PMID- 15892986 TI - Enhanced serum cholesterol and triglyceride levels in bulimia nervosa: relationships to psychiatric comorbidity, psychopathology and hormonal variables. AB - Increased levels of cholesterol have been reported in patients with bulimia nervosa (BN), but all but one of the published studies were performed on non fasting subjects, which limits the interpretation of this finding. Moreover, the relationships between serum lipids and comorbid psychiatric disorders or bulimic psychopathology have scarcely been investigated. We measured serum levels of total cholesterol, triglycerides, glucose, 17beta-estradiol and thyroid hormones in 75 bulimic women and 64 age-matched healthy females after an overnight fast. Compared with healthy women, bulimic patients exhibited significantly enhanced serum levels of cholesterol and triglycerides, but similar values of glucose, 17beta-estradiol, FT3 and FT4. No significant differences emerged in these variables between patients with or without comorbid depression, borderline personality disorder or lifetime anorexia nervosa. Circulating cholesterol was positively correlated to the patients' drive for thinness, ineffectiveness, enteroceptive awareness and impulse regulation sub-item scores of the Eating Disorder Inventory-2. These findings confirm that BN is associated with increased levels of serum lipids. This alteration may be involved in the pathophysiology of certain psychopathological characteristics of BN and cannot be explained by the co-occurrence of other psychiatric disorders. PMID- 15892987 TI - Suicide mortality in Southern Italy: 1998-2002. AB - The purpose of the study was to describe the deaths by suicides that occurred in an area of Southern Italy in the period 1998-2002. Data were obtained from death records, and population estimates for each year were provided by the National Institute of Statistics. A total of 367 suicides were retrieved. The number of suicides was stable for the 5 years recorded, with the highest value for 1998, ranging from 1.4% for ages 0 to 14 to 25.1% for ages 25 to 44 years. Age-specific rates of suicide showed that the highest rates were in those 75 to 84 years old. Rates are always higher in males, with a tendency to stability or decrease in all age groups in the years observed, except for males in the 45- to 64-year-old age range, in whom an increase from 5.71 to 7.28 was observed. The suicides increased proportionally with age, in those with the lowest level of education (53.3%) and among retirees (46.5%). Hanging/suffocation (44.1%) and jumping from high places (23.2%) were the most frequently used methods of suicide. Males had higher standardized suicide rates ranging from 5.4 per 100,000 in 1999 to 7.7 in 1998, whereas in females the range was 1.4 per 100,000 in 2001 to 2.4 in 1998. The study shows that suicide rates in Southern Italy are lower compared with national trends. PMID- 15892988 TI - Association between cigarette smoking and depressive symptoms among military medical students in Turkey. AB - An association has been documented between two important health concerns, smoking and depression. A cross-sectional study was carried out to explore whether this relationship exists in a sample of Turkish military medical undergraduates. Of a total of 779 military medical undergraduates, 690 agreed to participate in the study. The students completed a self-administered questionnaire including the Turkish adaptation of the Beck Depression Inventory (BDI) and gave a smoking history. It was determined that the smokers among medical students were 2.2 times more likely to have depressive symptoms than nonsmokers. This result extends the significant association between smoking and depressive symptoms in the literature to a sample of Turkish military medical undergraduates. PMID- 15892989 TI - Increased reproductive toxicity of landfill leachate after degradation was caused by nitrite. AB - Leachate from the landfill Lindbodarna was suspected to cause reproductive effects on fish in a Swedish lake, called Molnbyggen. The acute toxicity of this landfill leachate is caused by ammonia. In the present study the acute and chronic toxicity of the leachate from the landfill was tested with Ceriodaphnia dubia before and after treatment, either with (inoculated) or without addition of microorganisms from activated sludge, in both 2000 and 2001. On both occasions, the acute toxicity decreased after treatment, more rapidly with inoculum than without, and the cause of the decrease was mainly explained by decreasing concentrations of ammonia. However, the chronic toxicity decreased after treatment with inoculum but increased after treatment without inoculum. Therefore, we performed a series of acute and reproductive tests with ammonia, nitrite and nitrate on C. dubia, and the 24-h EC50s were 1.0, 2.7 and 59 mM, respectively, which are consistent with literature data. However, the chronic toxicity of these compounds gave quite a different picture with 8-day EC50s for reproduction of 3.0 mM for ammonia, 0.016 mM for nitrite and 1.5 mM for nitrate. Thus, the acute-chronic ratios for these compounds were 0.33 for ammonia, 170 for nitrite and 39 for nitrate. These findings show that reproduction is more sensitive than survival for both nitrite and nitrate, and that nitrite is the more hazardous of the two. This implies that the chronic and reproductive toxicity of nitrite and nitrate on zooplankton may in fact increase effects of eutrophication. In this study the toxicity of the fresh leachate was dominated by ammonia, but after treatment the contribution of nitrite increased, and especially the chronic toxicity of the treated landfill leachate was dominated by nitrite toxicity. PMID- 15892990 TI - The effect of high pH on ion balance, nitrogen excretion and behaviour in freshwater fish from an eutrophic lake: a laboratory and field study. AB - Slapton Ley is a freshwater hyper-eutrophic lake of two basins connected by a narrow channel. One part of the lake experiences summer blooms of cyanobacteria and poor water quality, including elevated water pH (maximum pH recorded=10.54), the other part is shaded by reed beds, and remains clear and neutral. This study used laboratory and field physiological measurements together with radio-tracking to investigate the potential impacts of alkaline pH on the physiology and behaviour of fish from Slapton Ley. Exposure of perch (Perca fluviatilis) from Slapton Ley to pH 9.50 water in the laboratory caused an immediate inhibition of sodium uptake and ammonia excretion to 34 and 32% of control levels, respectively. Net sodium balance recovered by day 3 of exposure whereas ammonia excretion only partially recovered to 60-70% of the control value from 8 h onwards. Urea excretion did not increase as a result of high pH exposure. Fish from the alkaline part of the lake (pH 9.90) had almost three-fold greater plasma ammonia compared to fish from neutral waters, indicating a pronounced disruption of ammonia excretion in the field. There was no significant disturbance to plasma sodium, chloride or total protein in fish sampled from the alkaline water of Slapton Ley. The radio-tracking provided no evidence of adult perch and pike (Esox lucius) trying to seek refuge from the alkaline conditions, despite having access to adjacent parts of the lake with neutral pH. It seems likely that there are advantages (e.g. better foraging, less predation) of withstanding the high pH conditions that outweigh the benefit of moving into more pH neutral parts of the lake. PMID- 15892991 TI - The human lipid regulator, gemfibrozil bioconcentrates and reduces testosterone in the goldfish, Carassius auratus. AB - Human and veterinarian pharmaceuticals have been detected in the aquatic environment for a number of years, but the potential for biological effects in exposed aquatic organism is only now being reported. The lipid regulator, gemfibrozil (GEM) is detected at microg/L concentrations in domestic wastewater and ng/L concentrations in surface waters. We investigated the uptake of GEM in goldfish (Carassius auratus) over a 96 h time period by measuring GEM in blood plasma using LC-MS/MS. Results indicated that GEM can be taken up from water through the gills. In goldfish exposed to GEM by a single intraperitoneal injection, concentrations of GEM in the blood plasma declined rapidly over 96 h post-injection, with a half-life estimated at approximately 19 h. Exposure of goldfish to waterborne GEM at an environmentally relevant concentration over 14 days resulted in a plasma bioconcentration factor of 113. In goldfish exposed to aqueous concentrations of GEM for 96 h or 14 days, plasma testosterone (T) was reduced by over 50% in fish from all treatments. As a possible mechanistic explanation for the observed reduction in T, levels of steroid acute regulatory (StAR) protein transcript in goldfish testes were assessed by RT-PCR. StAR protein is involved in the transport of cholesterol from the outer to the inner mitochondrial membrane for transformation by the first enzyme in steroidogenesis. After exposure to GEM for 96 h, a 50% decrease in StAR mRNA levels was observed in goldfish. Gonadal StAR mRNA levels were not affected in the 14 days exposure, indicating that the observed decreases in plasma testosterone were not solely due to impaired delivery of cholesterol to the inner mitochondrial membrane. Our results demonstrate that exposure to environmental levels of GEM leads to bioconcentration of the drug in plasma and the potential for endocrine disruption in fish. PMID- 15892992 TI - Genotoxic effects of selected biocides on RTG-2 fish cells by means of a modified Fast Micromethod Assay. AB - A sensitive in vitro assay for detecting DNA damage in RTG-2 cells culture is described. This assay employs a dye, PicoGreen double stranded DNA (dsDNA) quantitation reagent, which becomes intensely fluorescent upon binding nucleic acids. The assay includes a simple and rapid 50-min sample lysis in the presence of EDTA, SDS, and high urea concentration at pH 10, followed by time-dependent DNA denaturation at pH 11.6 after NaOH addition. The time course and the extent of DNA denaturation are followed in a microplate fluorescence reader at room temperature for less than 1h. Comparative studies between suspension and fixed RTG-2 cells indicated that it is possible to apply this methodology in both cases with good results. Neutral red assay was used for to determine the cellular viability when RTG-2 cultures were exposed to tetrakis(hydroxymethyl) phosphonium chloride (THPC) and benzalkonium chloride (BC), as biocides used in the disinfection of cooling towers. The results obtained by neutral red assay indicate IC(50(48)) values of 0.017 (0.011-0.028) and 2.71 (1.91-3.86) mg/L for tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. DNA damage has been evaluated for both disinfectants in RTG-2 culture, by exposure to 1/10-, 1/25-, 1/50-, and 1/100-IC(50(48)) value, and the results obtained indicate a strain scission factor (SSF) of 0.126+/-0.014, 0.181+/-0.014, 0.217+/-0.013, and 0.245+/-0.013 in cell suspensions, and 0.077+/ 0.019, 0.107+/-0.014, 0.151+/-0.014, and 0.202+/-0.015 in attached cells for tetrakis(hydroxymethyl) phosphonium chloride; while the SSF values for benzalkonium chloride are 0.023+/-0.009, 0.033+/-0.017, 0.068+/-0.012, and 0.088+/-0.015 in cell suspensions, and 0.033+/-0.010, 0.044+/-0.011, 0.080+/ 0.009, and 0.093+/-0.010 in attached cells. Thus, the assay proposed in this study has made it possible to show DNA damage in RTG-2 cells when exposed to 0.2(1/100 IC(50(48))) and 300(1/10 IC(50(48))) Hg/L of tetrakis(hydroxymethyl) phosphonium chloride and benzalkonium chloride, respectively. The results obtained indicate that the Fast Micromethod Assay, applied on RTG-2 cell line cultures, is a fast and sensitive method for the early DNA damage detection in the aquatic environment. PMID- 15892993 TI - Toxicity of selenite in the unicellular green alga Chlamydomonas reinhardtii: comparison between effects at the population and sub-cellular level. AB - The toxicity of selenium in aquatic ecosystems is mainly linked to its uptake and biotransformation by micro-organisms, and its subsequent transfer upwards into the food chain. Thus, organisms at low trophic level, such as algae, play a crucial role. The aim of our study was to investigate the biological effects of selenite on Chlamydomonas reinhardtii, both at the sub-cellular level (effect on ultrastructure) and at the population level (effect on growth). The cells were grown under batch culture conditions in well-defined media and exposed to waterborne selenite at concentrations up to 500 microM; i.e. up to lethal conditions. Based on the relationship between Se concentration and cell density achieved after a 96 h exposure period, an EC(50) of 80 microM with a 95% confidence interval ranging between 64 and 98 microM was derived. No adaptation mechanisms were observed: the same toxicity was quantified for algae pre contaminated with Se. The inhibition of growth was linked to impairments observed at the sub-cellular level. The intensity of the ultrastructural damages caused by selenite exposure depended on the level and duration of exposure. Observations by TEM suggested chloroplasts as the first target of selenite cytotoxicity, with effects on the stroma, thylakoids and pyrenoids. At higher concentrations, we could observe an increase in the number and volume of starch grains. For cells collected at 96 h, electron-dense granules were observed. Energy-dispersive X-ray microanalysis revealed that these granules contained selenium and were also rich in calcium and phosphorus. This study confirms that the direct toxicity of selenite on the phytoplankton biomass is not likely to take place at concentrations found in the environment. At higher concentrations, the link between effects at the sub-cellular and population levels, the over-accumulation of starch, and the formation of dense granules containing selenium are reported for the first time in the literature for a phytoplankton species after exposure to selenite. PMID- 15892994 TI - Sex and low-level sampling stress modify the impacts of sewage effluent on the rainbow trout (Oncorhynchus mykiss) immune system. AB - The objective of the present study was to investigate the influence of chronic exposure to municipal sewage treatment effluent at environmentally relevant concentrations on immune parameters in rainbow trout (Oncorhynchus mykiss), including the assessment of potential differences in reactivity between sexually mature male and female fish. Trout were exposed to 1.5 and 15% (v/v) secondary treated municipal sewage effluent for 32 weeks. Fish were injected intra peritoneally either with inactivated Aeromonas salmonicida to simulate an infection or with PBS as control for this immune challenge 6 weeks prior to sampling. Exposure to effluent resulted in a decrease in A. salmonicida-specific serum antibody level and blood lymphocyte numbers in mature females, but not in male fish. Injection of A. salmonicida resulted in enhanced serum lysozyme activity in mature male trout, which were not exposed to effluent. This stimulating effect of A. salmonicida could not be found in effluent-exposed trout, again potentially revealing a suppressive effect of the effluent. An influence of sampling fish on two consecutive days was observed in many immune parameters, most likely reflecting handling stress. Leucocyte and lymphocyte numbers in peripheral blood were consistently lower in male and female fish on the second sampling day. Phagocytosis in head kidney macrophages from male trout was also influenced by sampling day, whereby a stimulation of this reaction occurred on the second day of sampling. Liver mixed function oxygenase activity was found to be enhanced in mature male trout exposed to 15% effluent. In conclusion, the study showed, that exposure to sewage treatment plant effluent, in surface water relevant concentrations, can lead to potentially adverse effects on selected immune reactions in rainbow trout. However, this study also demonstrated that both handling stress and the sex of mature fish have distinct influences on the immune response detected in male and female fish and are likely to influence measured immune parameters to the extent that subtle effluent induced changes may be difficult to detect. PMID- 15892995 TI - Marine toxicity assessment of imidazolium ionic liquids: acute effects on the Baltic algae Oocystis submarina and Cyclotella meneghiniana. AB - Interest in ionic liquids for their potential in different chemical processes is increasing, as they are claimed to be environmentally benign and are very good non-volatile solvents for a wide range of applications. With regard to their physical and chemical characteristics, the properties of ionic liquids can be modified over a wide range because the cation's fine structure and the anion's identity can be altered. Since millions of ion combinations are possible it is of the highest importance to outline rational guidelines to develop technologically suitable but also environmentally harmless ionic liquids. This paper presents the results of a preliminary assessment of the toxicity of selected imidazolium ionic liquids towards marine algae. The selection of chemical entities was based on the t-SAR approach (thinking in terms of structure-activity relationships) focusing on the length (C(2) 5')-3'-O acetyl-5-fluoro-2'-deoxyuridine (dC(pam)-5-FdU(Ac), N4-palmitoyl-2',3' dideoxycytidylyl-(5' --> 5')-3'-O-acetyl-5-fluoro-2'-deoxyuridine (ddC(pam)-(5' - > 5')-5-FdU(Ac), 5-fluoro-2'-deoxyuridylyl-(3' --> 5')-5-fluoro-N4-hexadecyl-2' deoxycytidine (5-FdU-5-FdC(hex)), and of the new liponucleotide 1-O-octadecyl-rac glycerylyl-(3 --> 5')-5-fluoro-2'-deoxyuridine (Oct1Gro-(3 --> 5')-5-FdU). The anticancer activities of these prodrugs are comparable to those of 5-FdU and the tumor specificities are modulated by their structures. The highest cytotoxic activity being even superior to 5-FdU was expressed by the dimer 5-FdU-5 FdC(hex). PMID- 15893024 TI - Development of a prostaglandin D2 receptor antagonist: discovery of a new chemical lead. AB - A series of N-(p-alkoxy)benzoyl-5-methoxy-2-methylindole-3-acetic acids and N-(p butoxy)benzoyl-2-methylindole-4-acetic acid were discovered as new chemical leads for a prostaglandin D2 (PGD2) receptor antagonist. Most of them exhibited PGD2 receptor binding and blocked cyclic adenosine 3',5'-monophosphate (cAMP) formation in vitro. In particular, 2-methylindole-4-acetic acid analog 1 showed markedly increased receptor affinity and cAMP antagonist activity. Chemistry and structure activity relationship (SAR) data are also presented. PMID- 15893025 TI - [New diagnostic criteria and measuring outcome in polymyositis and dermatomyositis]. PMID- 15893026 TI - [Aetiological spectrum of hyperferritinemia]. AB - Serum ferritin levels may be increased in many conditions: renal diseases, liver diseases, human immunodeficiency virus infection. The purpose of this study was to assess the aetiological spectrum of high serum ferritin levels in a 1200-bed university hospital, to compare our results with the data already published and to assess a potential association between aetiology and ferritin levels. PATIENTS AND METHODS: Patients with a serum ferritin level higher than 600 microg/l were retrospectively included between 15 November 2003 and 15 January 2004, and their medical records were reviewed. RESULTS: Ninety-eight patients (38 women and 60 men; median age: 59,5 years [19-92]) were recruited in departments of hepatology and gastroenterology (22%), haematology (14%) and internal medicine (18%). Diagnosis performed were: non-HIV systemic infections (23,8%), haematological diseases (16,1%), alcoholism (11,2%) and malignancies (9,8%). Dialysed chronic renal failure, liver diseases, haemochromatosis and systemic inflammatory diseases counted for 4.2 to 5.2% of cases. Serum ferritin level lied between 600 and 1000 microg/l for 50 patients, between 1000 and 1500 microg/l for 24, and over 1500 microg/l for 24. There was no significant difference between the three groups as regards the etiological distribution. DISCUSSION: In our study, chronic renal failure was not a major cause of high ferritin level: this is probably due to the current use of erythropoietin, which has decreased the use of blood transfusions. The two major aetiology of hyperferritinemia were non-HIV infections and malignancies. PMID- 15893027 TI - [Interest of performing systematically the protein profile in an internal medicine department. National survey and prospective study on 229 admissions]. AB - INTRODUCTION: Despite a wide national use, the usefulness of the protein profile has only been evaluated in a small number of studies, essentially in patients with unknown diagnoses. METHODS: We conducted a survey on 339 french internal medicine departments to evaluate how the protein profile was used in these services. Concomitantly we achieved a prospective study on 229 patients in our department, with a mean follow up of 9 months, to evaluate how did the protein profile influence the diagnosis process. RESULTS: We received 183 responses to our national survey: the protein profile was available in 110/183 (60%) departments with 94/110 (85%) using it during hospitalisation and 20/94 (21.3%) using it systematically. Among the 229 protein profile analysed in our department, 44 (19.2%) were considered useful with 20 (8.7%) of them allowing the diagnosis of a new pathology. If the profile had not been done systematically, the physicians of our department would have performed the profile in 102/229 (44.5%) cases, whereas seven (3%) useful profiles would not have been done. CONCLUSION: We think that the profile has a consistent interest in hospitalized patients with a known or unknown pathology but performing systematically such a test appears to be of limited benefit. PMID- 15893028 TI - [Bone mineral density and bone metabolism in spondylarthropathies]. AB - AIMS: To study the prevalence of osteoporosis in a group of patients with spondyloarthropathy and to investigate bone turnover markers and correlation between bone mineral density and the age at the beginning of the disease. PATIENTS AND METHOD: Patients with spondyloarthropathy as defined by New York and ESSG criteria. Bone mineral density was measured at the lumbar spine and hip with Hologic QDR 1000. Serum levels of osteocalcin, deoxypyridinoline, 25 vitamin D, creatinine and parathyroid hormone were measured. RESULTS: 50 patients were included in the study: 37 men, mean age 40,2+/-13,8 years. Vertebral osteopenia was observed in 34% while femoral osteopenia occurred in 40% of patients. Serum vitamin D was low in 70% of patients without parathyroid hormone or kidney function modification. Markers of bone turn over were increased in 29% of patients. There was no correlation between these biological markers and the bone mineral density. We observed a significative correlation (P=0,02) between the age at the beginning of the disease and the bone mineral density. CONCLUSION: Osteopenia is present in patients with spondyloarthropathy without any correlation with the bone turnover biological markers. We observed a significative correlation between the age at the beginning of the disease and bone mineral density. PMID- 15893029 TI - [Cardiac manifestations of idiopathic hypereosinophilic syndrome]. AB - BACKGROUND: The aim of our study is to describe the more common cardiac manifestations of idiopathic hypereosinophilic syndrome representing the major cause of mortality. MAIN POINTS: Current therapy consists of corticosteroid, hydroxyurea and interferon alpha. Recent publications confirm the activity of imatinib mesylate, a selective tyrosine kinase inhibitor, in patients with idiopathic hypereosinophilic syndrome. In cases with marked valvular compromise or with endomyocardial thrombosis or fibrosis, cardiac surgery can provide substantial benefits. PERSPECTIVES: A better understanding of the pathophysiology of this syndrome could lead to the development of new therapeutic agents. PMID- 15893030 TI - [Molecular basis in hereditary haemochromatosis]. AB - PURPOSE: Recent discoveries in molecular mechanisms of iron metabolism have changed the classical view of hereditary iron overload conditions. We present natural mutations in newly discovered genes and related phenotypes observed in patients with different form of haemochromatosis. CURRENT KNOWLEDGE AND KEY POINTS: Most haemochromatosis patients are homozygous for the C282Y mutation in the HFE gene. Ferroportin, TFR2, hemojuvelin and hepcidin mutations also cause iron overload. Recent data support the hypothesis that haemochromatosis should no longer be considered a monogenic disease but rather an oligogenic disorder. Several results suggest that haemochromatosis could result from digenic inheritance of mutations in HFE and HAMP. FUTURE PROSPECTS AND PROJECTS: Other modifier genes probably influence penetrance in C282Y homozygous patients. Such genes could enhance or reduce the phenotypic expression in various iron overload conditions. PMID- 15893031 TI - [Subacute meningoradiculomyeloencephalitis due to cryptococcosis infection]. AB - INTRODUCTION: If meningoencephalitis with or without mass lesion (granuloma or abscess) is the most common pattern of CNS cryptococcal infection, intramedullary involvement is very uncommon. EXEGESIS: The authors report an 70-year-old male with Hodgkin's disease treated by chemotherapy then corticosteroids because of pulmonary fibrosis who was presenting for eight days ago, an ataxia, pyramidal syndrome, and bradypsychy. Spinal MRI revealed a gadolinium T1 weighted homogeneous enhancing T4 level intramedullary lesion. CSF had showed 190 GB/mm3 of lymphomonocytes, increased protein level (2.28 g/l), decreased glucose level (1.5 mmol/l) and positivity for crytococcal antigen. Treatment by amphotericine B and flucytosine then fluconazole for six months was instituted and symptoms gradually improved. CONCLUSION: A cryptococcus infection must be searched by antigen in CSF in case of myelopathy isolated or associated with meningoradiculoencephalomyelopathy, specially in patients with a cellular immunodeficience. Antimycotic agents must be firstly used, surgery would be restricted to decompression if aggravation of disease and compressive effect on the adjacent structures radiologically (MRI) became evident. Prolonged treatment is necessary in case of immunodeficience. PMID- 15893032 TI - [Systemic capillary leak syndrome: two case reports]. AB - INTRODUCTION: Systemic capillary leak syndrome (SHCI) is a rare disease with poor prognosis, characterized by recurrent episodes of generalized edema and hypotension. EXEGESIS: Two patients a 41 and 34-year-old woman were admitted with a generalized edema and several episode of shock. One patient had muscular edema with compartment syndrome imposing the aponevrotomia of discharge in emergency. The association of elevated hematocrit, decreased albumin serum concentration confirmed the diagnosis of SHCI. Administration of intravenous fluids improved the clinical and biological symptomatology. CONCLUSION: These two new observations are comparable to the published cases. We didn't find dysglobulinemia whose existence is frequented. The treatment of shock requires macromolecules solutions. The preventive treatment remains empiric. PMID- 15893033 TI - [Disseminated form of Rosai-Dorfman disease. A case report]. AB - INTRODUCTION: Extranodal involvement in sinus histiocytosis with massive lymphadenpathy (SHML) or Rosai-Dorfman disease is common, seen in 43% of cases. We present a case of disseminated form of SHML with thyroid, renal, bone and lung involvement. EXEGESIS: A 53-year-old woman presented with cervical lymphadenopathy and a large palpable mass in the submandibular area. A thyroid nodule was palpable. Laboratory data showed an increase of the erythrosedimentation rate and a polyclonal hypergammaglobulinemia. Histopathological examination of a cervical lymph node biopsy showed typical features of SHML. CT scan showed enlargement mediastinal lymph node, mass of the right lung and infiltrative mass in the right renal hilium. Radiographs revealed lytic lesions in the right proximal tibia and left patella. Histopathology of a biopsy from the kidney mass and thyroidectomy displayed typical characteristics of SMHL. The patient was treated by steroids with a markedly regression of the kidney and renal masses. CONCLUSION: Multiple extranodal involvement in SMHL is rare, the prognosis may be poor when lesions are massive and involve vital organs. PMID- 15893034 TI - [Cardiac pseudotumor revealing Behcet's disease]. AB - INTRODUCTION: Cardiac thrombosis is a rare complication of Behcet's disease (BD), which may present as a cardiac tumor. Its discovery precedes, in half of the cases, the diagnosis of BD. The high mortality may be associated to postsurgical complications and/or an associated involvement of pulmonary arteries. CASE REPORT: We present the case of a 31 years old Caucasian French woman, with a history of venous thromboembolic disease, who had surgery after the discovery of a right ventricle tumor. That was an organised thrombus with endomyocardial fibrosis and a diagnosis of Behcet's disease was made after the surgery. The outcome was favourable under medical treatment associating corticosteroids, colchicine and antivitamin K (AVK), without relapse four years later. CONCLUSION: The discovery of an intracardiac mass in a young patient must evoke the diagnosis of cardiac thrombus and Behcet's disease, even in the absence of predisposing ethnic or geographic factor. PMID- 15893035 TI - [Optic neuropathy revealing invasive aspergillosis of ethmoid and sphenoid sinuses]. PMID- 15893036 TI - [Onycholysis due to docetaxel in breast cancer treatment]. PMID- 15893037 TI - [Early onset polymyalgia rheumatica]. PMID- 15893038 TI - [Community-acquired bacteremic cellulitis with methicillin resistant Staphylococcus aureus carrying Panton-Valentine leukocidin gene]. PMID- 15893039 TI - [Coexistent psoriasis and systemic lupus erythematosus. A case report]. PMID- 15893040 TI - [Snoring: an unusual symptom revealing sarcoidosis]. PMID- 15893041 TI - [Mediastinal synovial sarcoma in the elderly: a case report]. PMID- 15893042 TI - Continuity through change: the rhetoric and reality of health reform in New Zealand. AB - New Zealand, like most other developed economies, has struggled to establish the best way of organising and delivering publicly financed health care services. Before the 1990s, hospitals and some related services were planned and provided by regionally based, locally elected health boards. This system was replaced in 1993 with a quasi-market structure in which separate organisations were responsible for the purchasing and provision of services. This in turn was replaced in 2001 by a system of locally elected boards that is remarkably similar to that which existed in the 1980s. The change to and subsequent abandonment of the quasi-market structure implies major changes to the way that health services are organised in New Zealand and suggests policy U-turns in (at least) three key areas: from cooperation to competition (and back); from integration of the roles of purchaser and provider to contractual arrangements (and back); and from local decision-making to centralised decision-making (and back). The aims of this paper are to examine the depth of system change in practice and to consider the extent to which the stated goals of reformers have disguised the degree of continuity between reform eras. We conclude that simplistic distinctions between structural approaches often fail to capture salient influences upon decision-making. New Zealand has not, in fact, been "to market and back" as the rhetoric would have us believe. Key aspects of the health system have endured throughout the reform period and these arguably have more importance for system functioning and performance than the high-level structural changes. PMID- 15893043 TI - Developing a scale for measuring professional equity among Canadian physicians. AB - This paper reports on progress made in defining and measuring the concept of professional equity through the development of a summative measure of professional equity and three of its components: financial, intrinsic and recognition equity. The study sample consisted of a stratified sample of 8375 Canadian physicians with usable responses from 2749 (32.8%). Following preliminary components analysis, items were grouped into constructs. Reliability of the constructs was then determined using Cronbach's alpha and total inter-item correlations followed by confirmatory factor analysis. A summary scale using all 15 equity items yielded a reliability: Cronbach's alpha=0.86. The sub-scales reliabilities were: financial equity (Cronbach's alpha=0.91); intrinsic equity (Cronbach's alpha=0.86); and recognition equity (Cronbach's alpha=0.70). The professional equity measures reported are therefore capable of assessing different aspects of equity and represent an advance over more general effort reward scales or those that only measure the range of rewards. PMID- 15893044 TI - The impact of considering adaptation in health state valuation. AB - Patients with chronic health conditions often rate their quality of life (QoL) significantly higher than non-patients. One explanation for this discrepancy is that non-patients focus on the negative aspects of the onset of a condition, especially the early difficulties people face when they first experience a debilitating condition, without considering that patients can adapt to it over time. To test this hypothesis, we had 359 people perform person tradeoff (PTO) elicitations in an online survey, varying whether the treatment programs under consideration saved the lives of patients (a) with pre-existing paraplegia; or (b) who would experience new onset of paraplegia. Half of each group completed an "adaptation exercise" which encouraged them to consider their own ability to emotionally adapt to negative events in general and specifically to having paraplegia. The adaptation manipulation increased the value participants placed on pre-existing paraplegia (p=0.03) and on new onset paraplegia (p=0.05), relative to saving healthy lives. Moreover, the adaptation exercise dramatically reduced the differences between evaluations of pre-existing and new onset paraplegia to values within 2% of each other. Our findings suggest that asking non-patients to do an adaptation exercise before giving QoL ratings may help close the gap in ratings between patients and citizen non-patients. PMID- 15893045 TI - Maternal and child health and family planning service utilization in Guatemala: implications for service integration. AB - Does the utilization of modern maternal and child health (MCH) services influence subsequent contraceptive use? The answer to this question holds important implications for proposals which advocate MCH and family planning service integration. This study uses data from the 1995/6 Guatemalan Demographic Health Survey and its 1997 Providers Census to test the influence of MCH service utilization on individual contraceptive use decisions. We use a full-information maximum likelihood regression model to control for unobserved heterogeneity. This model produces estimates of the MCH effect, independent of individual women's underlying receptiveness to MCH and contraceptive messages. The results of the analysis indicate that the intensity of MCH service use is indeed positively associated with subsequent contraceptive use among Guatemalan women, even after controlling for observed and unobserved individual- , household- , and community level factors. Importantly, this finding holds even after controlling for the unobserved factors that 'predispose' some women to use both types of services. Simulations reveal that, for these Guatemalan women, key determinants such as age and primary schooling work indirectly through MCH service use to increase contraceptive utilization. PMID- 15893046 TI - Smoking behavior in persons with a schizophrenia-spectrum disorder: a qualitative investigation of the transtheoretical model. AB - Smoking rates among persons with schizophrenia are up to three times the rates of the general US population, and research has shown that it is difficult to design cessation programs for people with schizophrenia that take into account their various cognitive and social deficits. More research is needed on the attitudes and priorities of people with schizophrenia in order to design and implement effective smoking cessation programs. Additionally, more research should be conducted with first-episode psychosis and chronic schizophrenia patients to investigate possible differences between these two groups. The purpose of this study, conducted in Atlanta, USA, was to use qualitative methodology to assess the Transtheoretical Model (TTM) in the context of smoking behavior in a sample of participants with schizophrenia-spectrum disorders. Data were obtained via interviews with 12 participants with either first-episode or chronic schizophrenia-spectrum disorders who smoked cigarettes. Differences between the two subsets of the sample were assessed. Results clustered into the following prevalent themes: (1) pros and cons of smoking; (2) beliefs about smoking cessation; (3) external influences on smoking and quitting; and (4) negative attitudes toward nicotine replacement therapies (NRT). Findings indicate that the majority of participants were in the precontemplation stage of quitting smoking, and that the primary advantages of smoking for this sample were relief from anxiety and negative symptoms. Important differences were found between chronic and first-episode participants in the areas of readiness-to-quit and beliefs about smoking cessation. Other findings indicate a lack of cessation programs offered to this sample, and overall negative attitudes toward NRT. Future interventions should take into account the reported pros and cons of smoking in this population, as well as other beliefs and attitudes regarding smoking behavior. PMID- 15893047 TI - Effects of patients with bipolar, schizophrenic, and major depressive disorders on the mental and other healthcare expenses of family members. AB - Family members who live with patients with serious mental disorders incur increased healthcare expenses. A retrospective study measured these increased expenses using administrative data from a large Blue Cross Blue Shield health plan in the USA. Mental and other healthcare expenses of family members of patients with bipolar disorder, schizophrenia, or major depression were compared to those of control family members. Ordinary least squares and logistic regression were used to estimate differences. In comparison with control (families of two or more members without these psychiatric disorders), mental and other healthcare expenses per family member per month were increased for subject family members as follows: bipolar disorder, $8.85 (213%; P<0.0001) and $10.65 (7.4%; P<0.0001); schizophrenia, $4.03 (81%; P<0.0001) and $5.96 (4.2%; P<0.005); and major depression $8.24 (219%; P<0.0001) and $9.46 (6.5%; P<0.0001). Among other factors, older family members had a greater likelihood of using mental healthcare and higher levels of other healthcare; males were less likely than females to use mental healthcare but had higher levels of other healthcare, and managed forms of coverage were associated with higher levels of both mental and other healthcare. We conclude that living with a person with serious mental illness significantly increases healthcare expenses of family members, especially mental healthcare. Family members of patients with bipolar disorder and major depression experienced larger increases in expenses than family members of patents with schizophrenia, despite the fact that patients with schizophrenia were more seriously ill as reflected in the much higher mental healthcare expenses of these patients. PMID- 15893048 TI - Family burden in long-term diseases: a comparative study in schizophrenia vs. physical disorders. AB - This study explored burden and social networks in families of patients with schizophrenia or a long-term physical disease. It was carried out in 169 specialised units (mental health department, and units for the treatment of chronic heart, brain, diabetes, kidney, lung diseases) recruited in 30 randomly selected geographic areas of Italy. The study sample consisted of 709 key relatives of patients with a DSM-IV diagnosis of schizophrenia and 646 key relatives of patients with physical diseases. Each relative was asked to fill in the Family Problems Questionnaire (FPQ) and the Social Network Questionnaire (SNQ). In all selected pathologies, the consequences of caregiving most frequently reported as always present in the past 2 months were constraints in social activities, negative effects on family life, and a feeling of loss. Objective burden was higher in brain diseases, and subjective burden was higher in schizophrenia and brain diseases than in the other groups. Social support and help in emergencies concerning the patient were dramatically lower among relatives of patients with schizophrenia than among those of patients with physical diseases. In the schizophrenia group, both objective and subjective burden were significantly higher among relatives who reported lower support from their social network and professionals. The results of this study highlight the need to provide the families of those with long-term diseases with supportive interventions, including: (a) the management of relatives' psychological reactions to patient's illness; (b) the provision of information on the nature, course and outcome of patient's disease; (c) training for the relatives in the management of the patient's symptoms; and (d) the reinforcement of relatives' social networks, especially in the case of schizophrenia. PMID- 15893049 TI - "Like roulette": Australian women's explanations of gynecological cancers. AB - Drawing on data from interviews and other ethnographic research, we examine how Australian women from different ethnic and cultural backgrounds make sense of having gynecological cancer. Alternative explanatory models often co-exist in a single narrative, but there is significant consistency in the etiology of cancer among Australian-born and immigrant women. In acknowledging the unpredictability of cancer and the prognosis of particular disease, women contextualize their own experience as a matter of luck, outside their control or influence. Most often women relate their own occurrence of cancer to their social setting and relational factors rather than personal behavior, but at the same time, they acknowledge the interaction of external forces and individual factors, particularly in the case of diet and stress. Women can control diet to some extent, although many note the irony of having developed cancer even when they have eaten well. Stress, on the other hand, is largely considered as external to and beyond women's control. Women speak of stress as a characteristic of contemporary social life, as well as their own public and domestic, physical and emotional lives; for these women stress in any of these areas can create physical vulnerability that may result in cancer. Since women associate cancer with loss of control, the idea of cancerous cells out-of-control within their bodies operates as a metonym of women's views of themselves interpersonally and socially. PMID- 15893050 TI - Medical disorders of suicides in Australia: analysis using a multiple-cause-of death approach. AB - The impaired health of a person who has committed suicide is often suggested among the proximate causes of suicidal behaviour. The introduction in 1997 of multiple-cause-of-death coding by the Australian Bureau of Statistics provided an opportunity to examine health impairments recorded on the death certificates of suicides. Data for the quinquennium 1997-2001 revealed a high prevalence of mental and behavioural disorders, in particular among women and among young and adolescent suicides. Comparison of multiple causes of death attributed to those who died in accidents with those recorded as suicides revealed that of the chronic and terminal illnesses, HIV and cancer were probably the conditions likely to trigger suicidal action. PMID- 15893051 TI - Women's experience of intimate partner violence in Haiti. AB - This study examined individual, partner, and community characteristics associated with the occurrence of intimate partner violence among ever-married women of reproductive age, using data from the 2000 Haiti Demographic and Health Survey. Separate logistic regressions were analyzed to assess women's risks of experiencing emotional, physical and sexual violence and multiple forms of intimate partner violence in the past 12 months. Twenty-nine percent of women in the sample experienced some form of intimate partner violence in the past 12 months, with 13 percent having experienced at least two different forms of violence. Significant positive associations with all forms of violence were found for lack of completion of primary school, history of violence exposure in women's families of origin either through witnessing violence between parents while growing up or direct experience of physical violence perpetrated by family members, partner's jealousy, partner's need for control, partner's history of drunkenness, and female-dominated financial decision-making. Significant positive associations were found between men's physical abuse of children at the community level and women's risk of experiencing emotional and physical violence. Neighborhood poverty and male unemployment, number of children living at home, women's attitudinal acceptance of wife beating, and male-dominated financial decision-making were additional risk factors for sexual violence. Women's economic independence was a protective factor for emotional and physical violence, while relationship quality was protective for all forms of violence and multiple victimizations. PMID- 15893052 TI - Does pregnancy provide immunity from intimate partner abuse among Hong Kong Chinese women? AB - The level of intimate partner abuse that occurs in Hong Kong Chinese families and the type, nature, and severity of abuse are primary concerns of those who are engaged in efforts to reduce and control abuse. A total of 1200 postnatal women were recruited at a university-affiliated regional public hospital to participate in a study with a retrospective explorative comparative design. The women were identified as abused or non-abused using the Abuse Assessment Screen Questionnaire (AAS). The type, nature, and profile of intimate partner abuse were investigated by using the Revised Conflict Tactics Scale (CTS-2). The vulnerable factors and predictors of abuse during pregnancy were explored in their demographic, psycho-socio-economic-cultural, and obstetric aspects. Analysis of the data indicated that 134 (11.2%) of the women had experienced abuse during pregnancy. The majority of this abuse took the form of psychological aggression (79.1%), and nearly half (49.3%) of the abused women had experienced sexual abuse. The husband was the perpetrator in the majority of cases (82.1%). Women who were younger, born in Hong Kong, single or cohabiting, had poor socio economic status, a poor partner relationship, and an unplanned pregnancy were more likely to be abused based on multiple logistic regression models of various nature and type of abuse (p<0.05). The findings provide fundamental information for the prevention of intimate partner abuse among the Hong Kong population. The implications, limitations, and future research are discussed. PMID- 15893053 TI - What does skin color have to do with infant health? An analysis of low birth weight among mainland and island Puerto Ricans. AB - This study investigates the relationship between maternal skin tone and low birth weight among Puerto Ricans, a group with a complex ancestry and skin tones that range from very light to very dark. Using data from a representative sample of Puerto Rican mothers, we assess whether skin tone has different implications for low birth weight in three geographic areas (Puerto Rico; New York City; other eastern states). The analysis shows that skin tone is unrelated to low birth weight in Puerto Rico and New York City. However, in the other eastern states in our sample, mothers with dark skin have a high risk of bearing a low birth-weight infant, relative to mothers with light skin. We interpret our findings in light of differences in the social meaning of phenotypic differences across locales. PMID- 15893054 TI - Visualising abortion: emotion discourse and fetal imagery in a contemporary abortion debate. AB - This paper presents an analysis of a recent UK anti-abortion campaign in which the use of fetal imagery--especially images of fetal remains--was a prominent issue. A striking feature of the texts produced by the group behind the campaign was the emphasis given to the emotions of those viewing such imagery. Traditionally, social scientific analyses of mass communication have problematised references to emotion and viewed them as being of significance because of their power to subvert the rational appraisal of message content. However, we argue that emotion discourse may be analysed from a different perspective. As the categorisation of the fetus is a social choice and contested, it follows that all protagonists in the abortion debate (whether pro- or anti abortion) are faced with the task of constructing the fetus as a particular entity rather than another, and that they must seek to portray their preferred categorisation as objective and driven by an 'out-there' reality. Following this logic, we show how the emotional experience of viewing fetal imagery was represented so as to ground an anti-abortion construction of the fetus as objective. We also show how the arguments of the (pro-abortion) opposition were construed as totally discrepant with such emotions and so were invalidated as deceitful distortions of reality. The wider significance of this analysis for social scientific analyses of the abortion debate is discussed. PMID- 15893055 TI - Boundary breaches: the body, sex and sexuality after stoma surgery. AB - People with limited or no bladder or bowel control, who have had a stoma to manage elimination, have a particular awareness of the proximity of the sites of pleasure and excretion. Drawing on interviews and related ethnography conducted in Australia from 1998 to 2001, this paper explores how men and women with permanent continence problems negotiate their sexuality around their bodily unreliability. Pleasurable sex, idealized, involves losing control. People who are incontinent or rely on a stoma, however, must monitor their bladder and bowel, disguising the stoma and bag and controlling their body in sex as in other circumstances. The need to negotiate bodily boundaries with established partners, or to disclose to new sexual partners, results in self-consciousness and social unease, and people need to reconstruct notions of privacy and dignity so that breaches in bodily control do not undermine the sexual relationship. For many, the stoma undermines self-esteem and body image, while its management confuses the status of the individual as "normal" and the partner as carer or lover. PMID- 15893056 TI - Storylines of research in diffusion of innovation: a meta-narrative approach to systematic review. AB - Producing literature reviews of complex evidence for policymaking questions is a challenging methodological area. There are several established and emerging approaches to such reviews, but unanswered questions remain, especially around how to begin to make sense of large data sets drawn from heterogeneous sources. Drawing on Kuhn's notion of scientific paradigms, we developed a new method-meta narrative review-for sorting and interpreting the 1024 sources identified in our exploratory searches. We took as our initial unit of analysis the unfolding 'storyline' of a research tradition over time. We mapped these storylines by using both electronic and manual tracking to trace the influence of seminal theoretical and empirical work on subsequent research within a tradition. We then drew variously on the different storylines to build up a rich picture of our field of study. We identified 13 key meta-narratives from literatures as disparate as rural sociology, clinical epidemiology, marketing and organisational studies. Researchers in different traditions had conceptualised, explained and investigated diffusion of innovations differently and had used different criteria for judging the quality of empirical work. Moreover, they told very different over-arching stories of the progress of their research. Within each tradition, accounts of research depicted human characters emplotted in a story of (in the early stages) pioneering endeavour and (later) systematic puzzle-solving, variously embellished with scientific dramas, surprises and 'twists in the plot'. By first separating out, and then drawing together, these different meta narratives, we produced a synthesis that embraced the many complexities and ambiguities of 'diffusion of innovations' in an organisational setting. We were able to make sense of seemingly contradictory data by systematically exposing and exploring tensions between research paradigms as set out in their over-arching storylines. In some traditions, scientific revolutions were identifiable in which breakaway researchers had abandoned the prevailing paradigm and introduced a new set of concepts, theories and empirical methods. We concluded that meta-narrative review adds value to the synthesis of heterogeneous bodies of literature, in which different groups of scientists have conceptualised and investigated the 'same' problem in different ways and produced seemingly contradictory findings. Its contribution to the mixed economy of methods for the systematic review of complex evidence should be explored further. PMID- 15893057 TI - Managing constraint: the experience of people with chronic pain. AB - This study describes the experience of people with chronic pain. Using the method of grounded theory, 29 chronic pain sufferers were interviewed at an outpatient pain clinic. A model depicting the basic social psychological process of maintaining a normal life through constraint was developed. This process revolved around people's perception of the constraints imposed by pain: bodily constraint (constraint on the body and its relationship to the environment); activity constraint (the constraint on what people could do); and identity constraint (the constraint on what people could be). The degree to which pain had challenged what people had previously accepted as 'normal' was illustrated through their evaluation of the impact of pain. The conclusion of this process of evaluation reflected how people coped with the constraints of pain-whether they were assimilated, accommodated, confronted or subverted. In assimilation, the constraints were absorbed and normal life maintained. In accommodation, the constraints were accepted and normal life re-defined. In confrontation, the constraints were rejected and pre-pain identities and activities pursued despite leading to increased pain levels. In subversion, attempts were made to retain pre pain identities, and although pain levels were minimized, activities were altered to a significant degree. The limitations imposed by pain often form the focus of people's coping efforts, rather than the pain per se. The desire to retain pre pain 'normal' lifestyles may underlie people's use of coping strategies that exacerbate pain intensity and pain-related disability. Future research needs to explore both the relationship between adjustment to pain and adjustment to the restrictions associated with ageing, and the role of body techniques and identity management in adjustment to pain in order to understand factors which may promote pain acceptance. PMID- 15893058 TI - 'Even if they ask you to stand by a tree all day, you will have to do it (laughter)...!': community voices on the notion and practice of informed consent for biomedical research in developing countries. AB - Ethical dilemmas in biomedical research, especially in vulnerable populations, often spark heated debate. Despite recommendations and guidelines, many issues remain controversial, including the relevance, prioritisation and application of individual voluntary informed consent in non-Western settings. The voices of the people likely to be the subjects of research have been notably absent from the debate. We held discussions with groups of community members living in the rural study area of a large research unit in Kenya. Discussions were facilitated by three research study vignettes outlining one field-based and two hospital-based studies being planned or taking place at the time. In addition to gathering general views about the aims and activities of the research unit, questions focused on whether consent should be sought for studies, and if so from whom (chiefs, elders, men/women, children), and on ascertaining whether there are any special concerns about the physical act of signing consent forms. The findings revealed the community's difficulty in distinguishing research from clinical investigations conducted in clinical settings. There was a spectrum of views regarding perceived appropriateness of consent procedures, in part because of difficulty in disentangling clinical from research aims, and because of other challenges to applying consent in practice. Debates between community members highlight the inadequacy of simplistic assumptions about community members' views on informed consent, and the complexity of incorporating lay opinions into biomedical research. Failure to appreciate these issues risks exaggerating differences between settings, and underestimating the time and resources required to ensure meaningful community involvement in research processes. Ultimately, it risks inadequately responding to the needs and values of those on whom the success of most biomedical research depends. Although compliance with community views does not necessarily make the research more ethical, it is argued that community opinions on local issues and practices should inform ethical decision making in health research. PMID- 15893059 TI - Inequities in healthcare seeking in the treatment of communicable endemic diseases in Southeast Nigeria. AB - This paper investigates the socio-economic inequities in healthcare seeking in the treatment of common communicable endemic diseases, with an emphasis on the use of primary health care (PHC) centres, where most endemic disease control activities take place. A questionnaire was used to collect information on occurrence of diseases and healthcare seeking from randomly selected households in four local government areas in Southeast Nigeria. Principal components analysis was used to create a socio-economic status (SES) index, which was divided into quartiles. The ratio of the values for the poorest quartile to that of the least poor quartile (bottom/top quartile) together with concentration indices for the variables under consideration was used as the measures of inequity. Logistic analysis was used to examine the determinants of use of PHC centres. The poorest quartile was more likely to use low-level providers (patent medicine dealers, shops, herbalists) and least likely to use the PHC centres. The concentration indices were -0.10, -0.06, -0.37, 0.11 and 0.04 for the use of herbalists, patent medicine dealers, community-health workers, PHC centres and hospitals, respectively. Also, the poorest quartile was more likely to lose person-days when ill. Logistic analysis showed that SES, availability of good services, proximity of the centres to the homes and polite health workers increased the use of the PHC centres. As such, improvement of quality of PHC services and improved geographic access could increase the overall use of PHC centres. Furthermore, in the long-term, a decrease in the amount of user fees, enhanced physical access and improved quality of services could decrease inequity in use of PHC centres and hospitals in the treatment of endemic diseases. The bottom/top quartile ratios and concentration indices produced similar results and hence both methods hence complement each other. PMID- 15893060 TI - From a relationship to encounter: an examination of longitudinal and lateral dimensions in the doctor-patient relationship. AB - Existing conceptualizations of the doctor-patient relationship provide little insight into this complex and perhaps now nonexistent "relationship" in the 21st century. Today, the word "relationship" as applied to the doctor-patient experience may be a misnomer--or at least an inappropriate description of the experience. One could ask, for example, if a person's most recent physician visit was more akin to their encounter with their last cab driver, or the person who sold them their last pair of shoes. After reviewing the 20th century theoretical conceptions of the doctor-patient relationship and describing the state of illness and health care delivery and policy in the United States, we develop a theoretical rubric for examining the 21st century physician-patient relationship. We argue that while patients should continue to be educated on how to use their time with physicians effectively and efficiently and physicians should continue to improve their communication with patients, we also argue that for policy purposes, it is not the physician or the patient that needs to change but rather the pressures and constraints of the organizational context within which the doctor-patient encounter takes place. PMID- 15893061 TI - The structure of patients' presenting concerns: the completion relevance of current symptoms. AB - This article uses conversation analysis to investigate the problem-presentation phase of 302 visits between primary-care physicians and patients with acute problems. It analyzes the social-interactional organization of problem presentation, focusing on how participants recognize and negotiate its completion. It argues that physicians and patients mutually orient to the presentation of current symptoms--that is, concrete symptoms presented as somehow being experienced in the here-and-now--as a locus of transition between the patient-controlled problem-presentation phase of the visit and the physician controlled information-gathering phase. This is a resource for physicians to distinguish between complete and incomplete presentations, and for patients to manipulate this distinction. PMID- 15893062 TI - Is what I have just a cold or is it dengue? Addressing the gap between the politics of dengue control and daily life in Villavicencio-Colombia. AB - In Colombia, dengue has been a target for public health interventions since the 1950s, with inadequate results. Furthermore, during the last few years in Colombia, the reported cases of dengue have increased. The social and cultural realities of the disease has been shown to be important and has not been considered when creating and implementing prevention and control programs. The paper draws on qualitative research of low- and high-income communities in the vicinity of Villaviecencio, South East Colombia to describe changing practices and direction of dengue prevention and control. It can be clearly observed from individuals' therapeutic itineraries and their perception of disease that public policies advertised in booklets and flyers and on television differ radically from people's everyday reality. This difference influences the success or failure of these policies. PMID- 15893063 TI - Susceptibility of West African Dwarf goats and WAD x Saanen crosses to experimental infection with Trypanosoma congolense. AB - West African Dwarf goats (WADs) and their Saanen crosses were experimentally infected with Trypanosoma congolense. No significant differences were found between trypanosome parasitaemia and antibody response of the crossbred and WAD goats. Neither the WAD goats nor the Saanen crosses were able to control the drop in PCV following trypanosome infection. The level of anaemia caused by the trypanosome infection was similar in the two breeds during the trial. Based on these findings, no difference in tolerance or susceptibility to T. congolense could be demonstrated between the WAD goats and their Saanen crosses. Although the weight of all goats increased during the trial, the crosses gained significantly more weight than the WAD goats. The trypanosome infection reduced the growth rate of both breeds, but this reduction was not statistically significant. Crossbreeding trypanotolerant WADs with trypanosusceptible Saanen goats might, therefore, be an effective means of increasing productivity. PMID- 15893064 TI - Identification of anti-babesial activity for four ethnoveterinary plants in vitro. AB - A commonly available Babesia caballi culture system was utilized for anti babesial screening of four commonly used ethnoveterinary plants, Rhoiscissus tridentata, Elephantorrhiza elephantina, Aloe marlothii and Urginea sanguinea, in vitro. Well-established B. caballi cultures were initially incubated with either imidocarb diproprionate and diminazene aceturate to validate the model, where after the studies were performed on the four plants. Effectivity was established as the degree of inhibition using a colour change method as well as by evaluating percentage parasitized cells on thin culture smears and calculating the degree of residual infectivity. The model was effective in demonstrating the in vitro efficacy of the well known anti-babesial drugs imidocarb and diminazene indicating an EC50 value of 0.08 and 0.3 microg/ml, respectively. Only the E. elephantina rhizomes acetone extracts were effective at a concentration of 100 microg/ml. It was also shown that the colour change method of evaluation was not very sensitive for determining activity of crude plant extracts. PMID- 15893065 TI - Seroprevalence of Neospora caninum in dairy and beef cattle with reproductive disorders in Japan. AB - Serum samples from 145 dairy and 65 beef cattle with reproductive disorders and 54 normally calving dairy cattle (controls) in Japan were tested for presence of Neospora caninum antibodies by use of an indirect fluorescent antibody test (IFAT, titer 1:200). Overall, seroprevalence of N. caninum was significantly higher (P < 0.001) in dairy cattle (20.0%, 29/145) than in beef cattle (1.5%, 1/65). In cattle which aborted, seroprevalence of N. caninum was significantly higher (P = 0.041) in dairy cattle (26.1%, 23/88, compared with controls (3.7%, 2/54)) than in beef cattle (5.0%, 1/20), indicating that neosporosis might be a more common problem in dairy cattle than in beef cattle in Japan. Seropositive cattle were 9.2 times more likely to abort compared to seronegative cows. Abortions associated with N. caninum seropositivity in this study were most frequently observed in the second trimester, and the mean gestational age of the fetuses aborted from seropositive dams was 5.7 months. In conclusions, N. caninum seems to be causing serious economic losses in the dairy industry in Japan. This is the first report on an objective comparison of seroprevalence of dairy and beef cattle with reproductive disorders in Asia. PMID- 15893066 TI - Infections in immunocompetent and immune-deficient mice with promastigotes of a North American isolate of Leishmania infantum. AB - Leishmania infantum, an etiologic agent of zoonotic visceral leishmaniasis, is widespread among foxhounds in the United States. Experimental infections with a North American isolate of L. infantum were evaluated using two inoculation routes in immunocompetent and immunosuppressed mouse strains. Groups of 2-5 interferon gamma gene knockout (IFN-gamma-KO) (BALB/c-Ifng), inducible nitric oxide synthase (NOS) gene knockout (iNOS-KO) (C57BL/6), B-cell-deficient (microMT) (C57BL/6), and BALB/c mice were intravenously (i.v.) or subcutaneously (s.c.) inoculated with various doses of promastigotes of the LIVT-1 strain of L. infantum. None of the mice developed clinical signs of leishmaniasis during the 8-9 weeks of the study. Promastigotes were cultured from spleens of all i.v.-infected mice by 3 days post culture. Spleens from s.c.-infected mice inoculated with greater than 1 x 10(6) parasites became culture positive 3-24 days post culture, but promastigotes were not cultured from mice infected with 1 x 10(5) or 5 x 10(5) LIVT-1 promastigotes. Histological lesions were prominent in the livers of i.v. infected mice but were mild to nonexistent in s.c. infection. Serological responses were low and transient determined by indirect fluorescent antibody testing in all groups. These results indicate that the i.v. route of infection is superior to the s.c. route in a mouse model of North American leishmaniasis and that mice lacking INF-gamma, iNOS or mice that are B-cell-deficient are not more susceptible to acute infection. PMID- 15893067 TI - Immune response to Neospora caninum in naturally infected heifers and heifers vaccinated with inactivated antigen during the second trimester of gestation. AB - The objective of this study was to compare the immune response to Neospora caninum in naturally infected heifers and heifers inoculated with a killed whole N. caninum tachyzoite preparation during the second trimester of gestation. Nine Holstein heifers were used in this study; three naturally infected heifers were born from seropositive dams, and six seronegative heifers were born from seronegative dams. Four seronegative heifers were subcutaneously vaccinated with a killed whole N. caninum tachyzoite preparation at weeks 13, 15 and 17 of gestation. A killed whole N. caninum tachyzoite preparation containing 45 mg of protein/5 ml dose was formulated with 70% of mineral oil adjuvant (13% consisting of Arlacel C, 85% Marcol 52 and 2% Tween-80). Similarly, two seronegative heifers (negative controls) were inoculated with mock-infected bovine monocytes in oil adjuvant. Humoral immune responses were tested by using an indirect fluorescent antibody test (IFAT) and an indirect enzyme-linked immunosorbent assay (ELISA) for detecting isotype specific antibodies. Cellular immune responses were assessed by lymphocyte proliferation test (LPT) and IFN-gamma production. N. caninum-specific antibody responses increased in immunized cattle by week 15 of gestation (mean reciprocal antibody titers 450+/-252), peaked at week 23 (mean 16,000+/-6400). Maximum antibody response in naturally infected heifers was observed at week 19 of gestation (mean: 3467+/-2810). Mean serum IFAT titers were significantly higher in immunized heifers compared with those in naturally infected heifers from weeks 17 to 25 (P < 0.05). Analysis of isotype specific antibodies in naturally infected heifers revealed a predominant IgG1 response in one heifer and a predominant IgG2 response in the other two. Similar titers of IgG1 and IgG2 occurred in immunized heifers. Control heifers remained seronegative throughout the study by IFAT and ELISA. Significant antigen-specific proliferation responses were only detected in naturally infected heifers in week 19 of gestation. Peripheral mononuclear blood cells (PMBC) from immunized animals produced IFN-gamma in similar concentrations to those of infected animals (P > 0.05). No abortion was seen in any experimental group; however, one calf from a vaccinated heifer died due to dystocia. All calves from vaccinated and control heifers were seronegative by IFAT at 6 months of age; in contrast, calves born from naturally infected heifers remained seropositive with titers > or = 200. Killed vaccine induced similar immune responses to those found in chronically, naturally infected cattle which did not abort; however, different immune pathways may be followed in vaccinated and natural infected heifers with differences in degree of protective immunity. PMID- 15893068 TI - Detailed analysis of an experimental challenge model for Leishmania infantum (JPC strain) in dogs. AB - In this study, disease progression after intravenous or subdermal infection of dogs with Leishmania infantum JPC strain was monitored. A challenge performed on 14 dogs via the intravenous route with 5 x 10(7) stationary phase promastigotes of the L. infantum JPC strain was 100% successful. During a follow up period of 1.5 years, several parameters were evaluated in order to find the most reliable disease markers. Parasite detection by culture and histology were found to be very sensitive (100%). Additionally, regular physical examination, serology and serum gamma-globulin levels were found to be useful parameters in the evaluation of disease severity and are recommended for inclusion in vaccination-challenge experiments. Although this intravenous challenge model has practical limitations, the data set confirms it is the best experimental model currently available for vaccine development. Two intravenously infected dogs were treated with corticosteroids for 5 months. This treatment was shown to enhance all aspects of a Leishmania infection. Five more dogs were infected by sub-dermal injection of promastigotes mixed with a proteophosphoglycan-matrix (PSG) secreted by Leishmania that assists in transmission and infection by sand fly bite. The resulting parasite burdens were low and the animals remained asymptomatic during a 2-year follow up period. However, this procedure did result in infection in 80% of the dogs and is appealing for future development as a natural challenge model in vaccine development. PMID- 15893069 TI - Experimental toxoplasmosis in red-legged partridges (Alectoris rufa) fed Toxoplasma gondii oocysts. AB - Thirty red-legged partridges (Alectoris rufa), 5-month-old, were orally inoculated with oocysts of the OV-51/95 strain of Toxoplasma gondii. Birds were distributed into five groups and received, respectively, 10 (group A, 4 birds), 50 (group B, 14 birds), 10(2) (group C, 4 birds), 10(3) (group D, 4 birds) and 10(4) (group E, 4 birds) oocysts. One partridge from group B and one from group E died suddenly of acute toxoplasmosis at 7 day after inoculation (DAI) with demonstrable T. gondii in several tissues. The rest of birds remained clinically normal until killed at 44, 58, 65, 72, 79 or 100 DAI. Brain, heart, liver and skeletal muscle from these partridges were bioassayed individually in mice; T. gondii was demonstrated in all these tissues, except in heart of three birds inoculated, respectively, with 10, 50 and 10(2) oocysts. Lesions were not seen in histologic sections of tissues from surviving partridges. These results suggest that red-legged partridges are resistant to clinical toxoplasmosis. PMID- 15893070 TI - Babesia spp. infection in Boophilus microplus engorged females and eggs in Sao Paulo State, Brazil. AB - Babesia spp. infections were investigated in Bos taurus x Bos indicus dairy cows and calves and in Boophilus microplus engorged female ticks and eggs. Blood samples and engorged female ticks were collected from 25 cows and 27 calves. Babesia spp. was detected in ticks by microscopic examination of hemolymph of engorged female and by squashes of egg samples. Cattle infection was investigated in blood thin smears and by DNA amplification methods (PCR and nested PCR), using specific primers for Babesia bovis and Babesia bigemina. Merozoites of B. bovis (3 animals) and B. bigemina (12 animals) were detected exclusively in blood smears of calves. DNA amplification methods revealed that the frequency of B. bigemina infection in calves (92.6%) and in cows (84%) and of B. bovis in calves (85.2%) and in cows (100%) did not differ significantly (P > 0.05). Babesia spp. infection was more frequent in female ticks and eggs collected from calves (P < 0.01) than from cows, especially in those which had patent parasitemia. Hatching rates of B. microplus larvae were assessed according to the origin of engorged females, parasitemia of the vertebrate host, frequency and intensity of infection in engorged female tick, and frequency of egg infection. Hatching rate was lower in samples collected from calves (P < 0.01) than from cows, and in those in which Babesia spp. was detected in egg samples (P < 0.01). PMID- 15893071 TI - Comparison between natural infection by Cryptosporidium sp., Giardia sp. in dogs in two living situations in the West Zone of the municipality of Rio de Janeiro. AB - In order to compare natural infection by Cyptosporidium sp. and Giardia sp. in clinically healthy dogs living in two situations (animal shelter and household pets), we conducted 166 faecal exams using the technique of centrifugation flotation in a sugar-saturated solution. Of the faecal samples, 94 came from shelter dogs and 72 from house pets. Eighty-two (49.4%) contained parasite eggs, cysts or oocysts. Of these, four (2.41%) contained Cryptosporidium sp. oocysts and 52 (31.33%) contained Giardia sp. cysts. There was no statistically significant difference between the origin of the dogs and Cryptosporidium sp. infection. Infection by Cryptosporidium sp. was not associated with the living conditions and sex. With respect to Giardia sp., we observed a statistically significant difference (p < 0.001) between the shelter dogs (45%) and the household pets 12.3%. There was no association of infection by Giardia sp. with age or sex. PMID- 15893072 TI - Serologic immunoreactivity to Neospora caninum antigens in dogs determined by indirect immunofluorescence, western blotting and dot-ELISA. AB - Neospora caninum, is a coccidian protozoan known as a major cause of bovine abortion and canine neuropathies. The aim of the present study was to develop a reliable and quick test to detect antibodies to N. caninum in dog sera. Sixty five serum samples from dogs, including 35 positive and 30 negative for N. caninum antibodies were used for standardization of the test. In parallel, immunoreactivity of the sera to Toxoplasma gondii antigens was investigated using a passive agglutination test. A dot-ELISA test, using soluble extract of N. caninum tachyzoites on nitrocellulose ester membranes, was developed and standardized. SDS-PAGE and complementary analysis of reactivity by Western blotting were used for the characterization of the immunoreactive fractions of all tested sera. The sensitivity and specificity of the dot-ELISA were 94 and 73%, respectively, compared to IFAT at a cut-off of 1:50, and 87 and 100% compared to IFAT at a cut-off of 1:25. Among the sera that tested positively for both IFAT and dot-ELISA, only 8.6% were reactive to T. gondii. The most immunoreactive fractions in Western blots were the 14-, 33-, 42- and 55 kDa bands, with percentages of 42, 60, 42 and 37%, respectively. The 60 kDa band showed a non-specific reaction in 43% of neosporosis-negative animals by both dot ELISA and IFAT. These results indicate that the dot-ELISA using N. caninum antigen present good sensitivity and specificity, and might be used as a screening test to detect antibodies to N. caninum in dogs. PMID- 15893073 TI - Detection of specific antibodies to Neospora caninum and Toxoplasma gondii in naturally infected alpacas (Lama pacos), llamas (Lama glama) and vicunas (Lama vicugna) from Peru and Germany. AB - Sera of an experimentally Neospora caninum infected llama and a non-infected control llama were used to establish an immunoblot, an ELISA and an IFAT to detect antibodies against N. caninum tachyzoites. Subsequently, serum samples collected from a total of 871 South American Camelids (SAC: Lama glama, Lama pacos, Lama vicugna) of two farms in Peru and from 32 SAC of a farm in central Germany were examined for antibodies against N. caninum and Toxoplasma gondii. Based on the recognition of specific bands in the immunoblot, sera of SAC from Peru were differentiated into N. caninum-positive (n = 18) and T. gondii-positive (n = 30) samples and into samples negative or inconclusive for both parasites. Using the immunoblot results as the reference, a modified version of the p38 ELISA and the IFAT were evaluated for detecting N. caninum antibodies in SAC sera. Applying a cut-off as determined by two graph-receiver operating characteristic analysis both, the ELISA and the IFAT, exhibited a sensitivity and specificity of about 95% in the SAC sera from Peru. Serological testing confirmed that SAC may become infected with N. caninum under field conditions in Peru. In addition to alpacas and llamas also 114 wild living vicunas had been examined for antibodies against N. caninum. However, only the alpacas and llamas but no vicunas were found N. caninum-positive. In contrast, T. gondii-seropositive animals were detected in all three SAC species. The lack of N. caninum seropositive vicunas indicates that in the study area in Peru wild canids might not serve as definitive hosts of N. caninum while for T. gondii a life cycle including wild felids is likely. On the German farm no N. caninum- but only T. gondii-seropositive SAC (n = 14) were detected. The seroprevalence of T. gondii infection was significantly higher in adult SAC (alpacas in Peru, llamas in Germany) than in crias (i.e. < 12 months old foals) indicating that the predominant route of infection is post natal. Since the present study was restricted to a few farms, the seroprevalences determined are not representative. However, our results confirm natural infections with N. caninum and T. gondii in SAC. Whether these infections are linked to any disease, e.g. reproductive losses, has to be clarified in further studies. PMID- 15893074 TI - Seroprevalence of T. gondii in sheep from Marrakech, Morocco. AB - Toxoplasma gondii is a ubiquitous intracellular protozoan parasite transmitted by food. Concerning this parasite, there are few studies done in Morocco. In this study, 261 sera from sheep intended for consumption in Marrakech were subjected to the Toxoplasma ELISA based serology test for the detection of anti-T. gondii specific IgG confirming a past infection. Of the total tested 72 (27.6%) sera were positive for IgG. This result shows that the seroprevalence approaches the world average and is similar to what is found in other cities of Morocco. This has prompted us to investigate other animal species in the region in order to evaluate the degree of contamination by this parasite as well as the potential risk incurred on consumption of their meat. PMID- 15893075 TI - Toxoplasma gondii infection in adult llamas (Lama glama) and vicunas (Vicugnavicugna) in the Peruvian Andean region. AB - The present study was designed to investigate Toxoplasma gondii infection in adult llamas (Lama glama) and vicunas (Vicugna vicugna) in the Peruvian Andean region, for which to date no information has been available. Serum samples from 43 llamas (L. glama) and 200 vicunas were tested by IFAT detecting titres of 1:50 or higher in 55.8% (33.9-70.9%) and 5.5% (2.8-9.6%), respectively. IFAT titres ranged from 1:50 to 1:6400. In order to avoid cross reactions with closely related coccidian parasites and to confirm the existence of T. gondii specific antibodies, IFAT positive sera from both ruminant species were also analysed by western blot. T. gondii specific antigens were recognised by IFAT positive sera, although different IFAT cut-off points could be selected for llamas (1:200) and vicunas (1:50) meaning seroprevalence of 44.2% (29.1-60.1%) and 5.5% (2.8-9.6%), respectively. Based on the frequency and intensity of tachyzoite antigen recognition, at least three immunodominant antigens with apparent molecular weights of 22-24, 30, and 38-40 kDa were detected, together with other minor protein fractions located in the 18-73 kDa range. This study documents for the first time the presence of T. gondii infection and reports the target T. gondii antigens in adult llamas and vicunas in Peru. PMID- 15893076 TI - Risk factors for canine echinococcosis in an endemic area of Peru. AB - An epidemiological study was conducted in a highland rural community in Peru to determine risk factors for canine echinococcosis caused by Echinococcus granulosus. Dogs were diagnosed using a coproantigen enzyme-linked immunosorbent assay (ELISA). Dog owners were interviewed prior to stool collection and asked for attitudes, practices and beliefs likely to be associated with local patterns of E. granulosus transmission. Univariate and multivariate analysis were used to determine odds ratios (OR) and their 95% confidence intervals (CI). The main risk factors found to be significantly associated with canine echinococcosis by univariate analysis were dog age (3-25 months) (OR, 5.14; CI, 1.7-15.7), female sex (OR, 4.3; CI, 1.4-13.3) and having been fed hydatid infected offal (OR, 2.9; 95% CI, 1.0-8.6). There was complete lack of knowledge about echinococcosis transmission. In addition to periodic dog treatment, control programs need to emphasize education of the human population to increase knowledge of parasite transmission and to change human practices associated with high rates of infection. PMID- 15893077 TI - The effect of grazing forage containing condensed tannins on gastro-intestinal parasite infection and milk composition in Angora does. AB - The objective of this study was to evaluate effects of the condensed tannin (CT) containing forage sericea lespedeza (sericea lespedeza (SL); Lespedeza cuneata; 15.2% CT), on fecal egg count (FEC), larval development (larvae/10 g of feces), worm burden and immune response compared with a crabgrass (Digitaria ischaemum)/Kentucky 31 tall fescue (Festuca arundinacea; control forage (CTF)) forage low in CT (0.32% CT) in grazing Angora does and their kids. Fifty worm free mixed-age does were randomly allocated to three treatments. One treatment (10 does; initial liveweight (LW) = 45+/-1.5 kg) entailed grazing of SL forage from April 25 to July 15, 2002 with a second treatment of CTF (20 does; initial LW = 43+/-1.4 kg) grazing during the same period. Does of the third treatment (20 does; initial LW = 44+/-1.4 kg) grazed a sward of SL for 2 weeks and then one of CTF for 2 weeks followed by alternating between the two pastures every 2-week rotational grazing (ROT). To gauge levels of infective larvae on pasture, three worm-free Angora kids (initial LW = 3.6+/-0.2 kg) were randomly selected as tracers. Tracers grazed for final 60 days and were euthanized for determination of worm burden. The immune response of does was measured by skin thickness reaction after the intradermal injection of 250 microg phytohemagglutinin (PHA). Mean FEC for SL and ROT were substantially lower (P < 0.01) than for CTF does (145, 329 and 894 eggs/g, respectively). The FEC for kids was lower (P < 0.05) for SL than for ROT and CTF (550, 2757 and 3600 eggs/g, respectively). Total fecal egg output (3.3, 6.0 and 26.9 x 10(5) eggs/day, respectively) and larval development (242, 263 and 792 larvae/10 g, respectively) were lower (P < 0.05) for SL and ROT than for CTF. Tracers grazing on SL had lower total worm burdens than ROT and CTF (P < 0.01). The immune response was higher (P < 0.01) for SL (4.9 mm) and ROT (6.0 mm) than for CTF (3.0 mm) at 12 h after injection of PHA. The packed cell volume (PCV) in does was higher (P < 0.01) for SL and ROT than for CTF (27, 26 and 23%, respectively). Does that grazed CT-containing forage had considerably lower milk somatic cell counts (SCC) than does grazing non-CT containing forage. In summary, grazing CT forages reduced FEC, larval development and worm burden, and also appeared to enhance immune response. The CT-containing forage SL reduced gastro-intestinal parasite infections of Angora does and kids. PMID- 15893078 TI - The use of an indirect Ostertagia ostertagi ELISA to predict milk production response after anthelmintic treatment in confined and semi-confined dairy herds. AB - An indirect Ostertagia ostertagi ELISA was used in late lactation milk samples from cows in confined and semi-confined dairy herds to determine if it could predict milk production response after endectocide treatment at calving. Holstein cows from 9 dairy farms from Prince Edward Island (PEI), 5 from central Nova Scotia and 16 from southern Ontario that were participating in a clinical trial of endectocide treatment around calving were used in this study. The cows were randomly treated with either eprinomectin pour-on endectocide or a placebo solution. Milk samples were obtained from cows late in the lactation before treatment was applied. These samples were tested for antibodies to gastrointestinal nematodes (GIN) using the indirect ELISA with the results expressed as optical density ratios (ODR). Production records were obtained from a computerized database of dairy herd improvement data. Pre-calving ODR showed a seasonal pattern. They were higher in the summer and fall and lower during the winter months. Older animals had higher pre-calving ODR values compared with younger cows. Similarly, cows from semi-confined herds had higher parasite antibody levels compared with cows from confined herds. The endectocide treatment did not affect the milk production response in the overall study population. However, the interaction effect between treatment and pre-calving ODR on milk production response after endectocide treatment was significant (P = 0.02), with some evidence of positive treatment response in cows with an ODR > 0.4. The relationship between pre-calving ODR and production response appeared to be quadratic rather than linear. PMID- 15893079 TI - Serological and molecular survey of Dirofilaria immitis infection in stray cats in Gyunggi province, South Korea. AB - The aim of this study was to carry out a serological and molecular survey for the presence of Dirofilaria immitis infection in stray cats using an ELISA kit and PCR assay. One hundred and fifty-five stray cats (77 females and 78 males) in Gyunggi province in South Korea, were used in this study. Four (2.6%) tested with the ELISA kit showed a positive reaction, and all positive samples by the ELISA kit showed a positive reaction by PCR analysis. No significant difference was observed between the male (2.6%) and female (2.6%) cat groups by ELISA kit. The positive rates for dirofilariosis were 2.8% in the 4-6-year-old group, and 18.2% in the > 6-year-old group by ELISA kit. With regard to the age element, older cats showed a higher prevalence of D. immitis infection in this study. A statistical analysis revealed that significant difference was observed in > 6 year-old group (p < 0.01). In conclusion, D. immitis infection in stray cats was present in Gyunggi province, although its incidence was low. Therefore, heartworm treatment and/or prophylaxis for stray cats captured are required in this area. PMID- 15893080 TI - Biological and DNA evidence of two dissimilar populations of the Rhipicephalus sanguineus tick group (Acari: Ixodidae) in South America. AB - In this work, the biology, mitochondrial DNA and fertility of hybrids from two strains of Rhipicephalus sanguineus, from Brazil and Argentina, were compared. Engorged larvae, nymphs and adults from Argentina weighed more and the engorgement period of adult females was significantly longer than those of their Brazilian counterparts, whereas adult female tick yield rate was higher for the Brazilian strain. High intraspecific divergence of mitochondrial DNA was detected between R. sanguineus from Brazil and Argentina. On the other hand, a strong genetic relationship was detected between European and Argentinean R. sanguineus populations while the Brazilian population appeared to be related to the African Rhipicephalus turanicus. Adult hybrid females laid eggs, which were mostly unviable, whereas a mean of more than 1400 larvae hatched per egg mass from pure Brazilian and Argentinean strains. These results showed that differences between these strains are greater than previously assumed and that the biosystematic status of R. sanguineus ticks from South America should be re-evaluated. Wide variations, such as these might account for the reported worldwide differences in biology and vector capacity of this species. PMID- 15893081 TI - A review of Melophagus ovinus (L.), the sheep ked. AB - The sheep ked Melophagus ovinus is a member of the parasitic Dipteran family Hippoboscidae; it was a widespread ectoparasite of sheep that was of veterinary importance until the introduction of effective pesticides. Since then it has become of less concern and may have become a relatively uncommon species through much of its former range. Changes in agricultural practice, both greater intensification and development of more extensive systems (including organic farming and conservation grazing), may allow resurgence and a review of existing knowledge of M. ovinus may be timely. This paper reviews the biology, annual population cycles, natural causes of mortality, veterinary and economic significance and control of, and host resistance to, M. ovinus. PMID- 15893082 TI - Fleas parasitizing domestic dogs in Georgia, USA: species composition and seasonal abundance. AB - Monthly flea collections were made from domestic dogs in Bulloch County, Georgia, USA from September 1996 to August 2004. A total of 2518 fleas belonging to 8 species were collected. The most common flea was the cat flea, Ctenocephalides felis (389 males and 1148 females), followed by the dog flea, Ctenocephalides canis (250 males and 285 females), a generalist/carnivore flea, Pulex simulans (106 males and 213 females), and a sticktight flea, Echidnophaga gallinacea (3 males and 89 females). Small numbers of rabbit-associated fleas (25 Cediopsylla simplex and 6 Odontopsyllus multispinosus) and rodent-associated fleas (3 Orchopeas howardi and 1 Polygenis gwyni), suggested that certain dogs had acquired these particular ectoparasites through hunting activities. Sex ratios of each of the five most frequently collected flea species were female-biased. Seasonally, C. felis, C. canis, and P. simulans, all showed distinct abundance peaks in late summer or autumn. PMID- 15893083 TI - An outbreak of abortions and high neonatal mortality associated with Trypanosoma evansi infection in dromedary camels in the Canary Islands. AB - Trypanosoma evansi was diagnosed for the first time in the Canary Islands (Spain) in 1998 in a dromedary camel. Seroprevalences of 4.8% up to 9% have been observed using different diagnostic methods. Affected animals have been treated but the dissemination of the disease is unknown. This article presents an outbreak of abortions and high neonatal mortality attributable to T. evansi infection in camels as well as the clinical assessment of the affected animals. The patients were diagnosed by routine checking (three pregnant animals), after abortion (five dams), or after delivered premature or weak calves (eight dams). At clinical examination, 2 out of 16 affected animals showed moderate signs of chronic form, particularly hyporexia and intolerance to exercise. The aborted fetuses were aged 6-8 months of gestation, approximately. The main laboratorial findings were regenerative anemia (haemolytic anemia), lymphocytic and monocytic leukocytosis, hyperproteinemia, hyperglobulinemia, hypoglycaemia, serum urea increased and serum iron decreased. Treatment using trypanocidal drug (Cymelarsan) resulted highly effective. Massive treatment would be recommended in the entire camel population in the Canary Islands (less than 2000 animals), as therapeutic or preventive measure, in order to control and to achieve an eventual eradication of the disease. PMID- 15893084 TI - Prevalence of antibodies to Toxoplasmagondii and Neosporacaninum in red foxes (Vulpesvulpes) from around the UK. AB - The purpose of this study was to establish the prevalence of antibodies to Toxoplasma gondii and Neospora caninum in a random sample of red foxes from around the UK. Lung fluid from over 500 foxes was examined using an indirect fluorescent antibody test. Reciprocal titres of specific antibodies to T. gondii or N. caninum ranged from < 1:16 to 1:1024. A titre of 1:128 or greater was deemed indicative of exposure to the parasite. One hundred and eleven (20%) of the 549 foxes tested were seropositive to T. gondii, and only five (0.9%) were seropositive to N. caninum. No correlation could be made between positive samples and geographical distribution, as sample numbers varied greatly between regions. The results of this study indicate that red foxes of the UK have more exposure to T. gondii than to N. caninum in their environment. PMID- 15893085 TI - Brief motivational intervention for adolescent smokers in medical settings. AB - This study evaluated the efficacy of using a brief motivational intervention to reduce smoking among adolescent patients treated in a hospital outpatient clinic or Emergency Department. Patients aged 14-19 years (N=85) were randomly assigned to receive either one session of motivational interviewing (MI) or standardized brief advice (BA) to quit smoking. The assessment and intervention were conducted in the medical setting proximal to the patient's medical treatment. Patients were proactively screened and recruited, and were not seeking treatment for smoking. Follow-up assessments were conducted at 1, 3, and 6 months post-intervention. Self-report data indicated that 7-day abstinence rates at 6-month follow-up were significantly higher in the MI group than in the BA group, but this difference was not confirmed biochemically. Self-reported smoking rate (average cigarettes per day) was significantly lower at 1, 3, and 6 months follow-up than it was at baseline. Cotinine levels indicated reduced smoking for both groups at 6 months, but not at 1 month. At 3-month follow-up, only those in MI showed cotinine levels that were significantly reduced compared to baseline. Findings offer some support for MI for smoking reduction among non-treatment-seeking adolescents, but overall changes in smoking were small. PMID- 15893086 TI - Difficulty in demonstrating a risk from drinking pattern in fourteen years of coronary heart disease morbidity and mortality: the Lung Health Study. AB - The health effects of a binge pattern of alcohol consumption have not been widely investigated. The objective of this study is to evaluate cardiovascular consequences of drinking eight or more drinks at a sitting and of usual drinking of alcohol among 3702 men in the Lung Health Study (LHS), a clinical trial where heavy drinkers were excluded from enrollment. Using a 14-year follow-up period, survival graphs were examined. Cox proportional hazards regressions were performed on time to first event for documented hospitalizations and deaths due to coronary heart disease (CHD). The upper two quartiles of usual drinking were protective against CHD in men [hazard ratios (HRs) 0.76 and 0.69] in adjusted models. When eight or more drinks per occasion was combined with models of usual drinking quartiles, its effect was not significant. The measure of eight or more drinks in these data appears to act as a surrogate for heavy drinking, and does not provide a suitable test of the effect of drinking pattern in men, due primarily to the exclusion of heavier drinkers at baseline. The alcohol effects in women in this study were not significant. PMID- 15893087 TI - Factors affecting American Indian adolescent tobacco use. AB - The present study merged problem behavior and social ecological theories to examine how mental health and environmental factors, including culture, were associated with American Indian youth tobacco use. A stratified random sample of 205 reservation and 196 urban American Indian adolescents living in a Southwestern area was interviewed in 2001. Two-thirds of the reservation youth and half of the urban youth in this sample reported lifetime tobacco use. Logistic regression showed that, when controlling for age and location, a mental health factor (substance abuse/dependence) and environmental factors (e.g., family members' mental health problems and peer misbehavior) were significant predictors of American Indian adolescent tobacco use. Cultural factors and location (reservation vs. urban) were not significant predictors of their tobacco use. Therefore, environmental and mental health factors should be assessed for and incorporated into tobacco use intervention and prevention plans for American Indian youth in both reservation and urban areas. PMID- 15893088 TI - Understanding the associations among education, employment characteristics, and smoking. AB - The current study examined the association between education level and smoking status in a community-based sample of working adults. Participants were enrolled at the University of Texas M.D. Anderson Cancer Center site of a cancer risk behavior reduction intervention delivered at the worksite. There was a strong educational gradient in smoking status. The prevalence of current smoking was almost threefold higher among individuals with 0.7 for the "Adverse effects" and self-efficacy scales, but was low (0.2-0.4) for self-change strategies, which probably reflects active use of these strategies in this sample. The translated scales performed adequately in most tests of construct validity. In particular, higher self-efficacy ratings predicted smoking cessation at follow-up, and a lower self-efficacy predicted relapse in baseline ex-smokers. CONCLUSION: The validity of the scales was maintained after translation in English. PMID- 15893094 TI - Comparison of the predictors of alcohol use and misuse among Han and aboriginal students in Taiwan. AB - The objective of this study was to compare the predictors of alcohol usage between the Han and aboriginal students in Taiwan. Results showed a significant gender difference for alcohol use and problematic drinking in both Han and aboriginal adolescents, with males having a higher prevalence of these problems than females in both groups. Aboriginal adolescents did not show gender differences in alcohol-related negative consequences. Male, paternal drinking, single-parent household, peer drinking, and peer relationships are the significant predictors for the Hans adolescents' problematic drinking. Gender, paternal drinking, and peer drinking are the significant predictors for aboriginal adolescents' problematic drinking. If the father drinks frequently, odds ratio is 6.48 likely for aboriginal adolescents to have problematic drinking. If peers drink frequently, odds ratio is 6.26 likely for aboriginal adolescents to have alcohol-related negative consequences. Frequent peer drinking was associated with a 6.26 odds ratio of have alcohol-related negative consequences. PMID- 15893095 TI - Analysis of baseline by treatment interactions in a drug prevention and health promotion program for high school male athletes. AB - This paper investigates baseline by treatment interactions (BTI) of a randomized anabolic steroid prevention program delivered to high school football players. Baseline by treatment interactions occur when a participant's score on an outcome variable is associated with both their pretreatment standing on the outcome variable and the treatment itself. The program was delivered to 31 high school football teams (Control=16, Treatment=15) in Oregon and Washington over the course of 3 years (Total N=3207). Although most interactions were nonsignificant, consistent baseline by treatment interactions were obtained for knowledge of the effects of steroid use and intentions to use steroids. Both of these interactions were beneficial in that they increased the effectiveness of the program for participants lower in knowledge and higher in intentions at baseline. PMID- 15893096 TI - Contributions of attachment style and perceived social support to lifetime use of illicit substances. AB - Research has shown insecure attachment style is associated with ineffective emotional regulation leading to maladaptive behaviors in adulthood. In the present study, we examined the association between attachment style and illicit substance use within a sample of adoptees (n=148). It was predicted that insecure attachment style would be associated with a higher incidence of lifetime illicit substance use and that perceived social support would mediate this association. Logistic regression analyses showed higher prevalence of illicit substance use among both insecure attachment groups as compared to the secure group. No difference was found between the two insecure types. Perceived social support was found to mediate the association between attachment style and illicit substance use for the insecure-preoccupied group only. The findings from the present study further implicate attachment style in the risk for illicit substance use, as well as preventions designed to identify those at risk for use. PMID- 15893097 TI - Cutting down substance abuse--present state and visions among surgeons and nurses. AB - BACKGROUND: Although substance abuse is variably common among surgical patients, detection of the problem and brief interventions are seldom undertaken. The aim of the present study was to assess surgeons' and nurses' activity in screening substance abuse among their patients and obstacles concerning intervention of patients with substance abuse. METHODS: Surgeons and nurses of The Department of Surgery, Tampere University Hospital were asked to predict if the surgical patients treated during a 24-h period (n=211) had a substance abuse disorder. This was compared to the patients' self-reports based on the Alcohol Use Disorders Identification Test (AUDIT) and questions about use of other substances. The surgeons and the nurses were also asked to complete a questionnaire related to interventions of patients with substance abuse; perceived knowledge, skills, attitudes and obstacles. RESULTS: Of the surgical patients, 47 (22%) proved to be substance abusers. Out of them, 23% were detected by surgeons and nurses. Of the surgeons (n=44) 83% and of the nurses (n=332) 84% perceived responsibility for intervention but claimed many obstacles. Nurses reported the lack of knowledge as the main obstacle. Of the nurses 18% reported enough knowledge to conduct the whole brief intervention compared to 34% of surgeons (p=0.021). The main obstacle reported by surgeons was the lack of time. CONCLUSIONS: Surgeons and nurses detect only minority of the substance abusing surgical patients. Lack of knowledge and time prevent intervening in patients' substance abuse. These results may be utilized when developing a program to better involve surgeons and nurses in detection and intervention of surgical patients' substance abuse. PMID- 15893098 TI - Comparison of drug use and psychiatric morbidity between prostitute and non prostitute female drug users in Glasgow, Scotland. AB - AIMS: To compare psychiatric morbidity between 176 female drug users with lifetime involvement in prostitution (prostitutes) and 89 female drug users with no involvement (non-prostitutes) in Glasgow, Scotland. METHOD: The Revised Clinical Interview Schedule (CIS-R) measured current neurotic symptoms. RESULTS: Prostitutes were more likely to report adult physical (OR 1.8) or sexual abuse (OR 2.4), to have attempted suicide (OR 1.7) and to meet criteria for current depressive ideas (OR 1.8) than non-prostitutes. Seventy-two percent of prostitutes and sixty-seven percent of non-prostitutes met criteria for a level of current neurotic symptoms likely to need treatment (CIS-R > or = 18). Being in foster care (OR 8.9), being prescribed medication for emotional problems in the last 30 days (OR 7.7), adult sexual abuse (OR 4.5), poly drug use in the last 30 days (OR 3.6) and adult physical abuse (OR 2.6) were significantly associated with a CIS-R score of > or = 18 for prostitutes using multiple logistic regression. CONCLUSIONS: Higher rates of adulthood abuse among prostitutes may explain the greater proportion of prostitutes than non-prostitutes meeting criteria for current depressive ideas and lifetime suicide attempts. PMID- 15893099 TI - Factors predicting 2-year retention in methadone maintenance treatment for opioid dependence. AB - Factors predicting 2-year retention in methadone maintenance and the impact of repeat treatment episodes on retention are examined. Data (n=9555 episodes) were drawn from a population-based treatment registry and analyses were performed using episode-weighted data. We estimated a binary logistic regression model with a duration of 730 days or greater as the dependent variable. The odds of remaining in treatment for 730 days or more increase with age and vary by region and provider type, but decrease with increasing number of treatment episodes. In comparison with other studies, these analyses show much higher rates of retention in methadone treatment but suggest that repeat episodes may not be as beneficial as existing research suggests. PMID- 15893100 TI - A psychometric evaluation of the Fagerstrom Test for Nicotine Dependence in PTSD smokers. AB - Rates of smoking among individuals with psychiatric conditions are greater than rates seen in the general population, yet little is known about the psychometric properties of commonly used nicotine dependence instruments among psychiatric smokers. This study examined the reliability, validity, and factor structure of the Fagerstrom Test for Nicotine Dependence (FTND) among psychiatric smokers. Results revealed that the FTND had good test-retest reliability, convergent validity, and discriminant validity. A factor-analytic examination converged on a two-factor solution, reflecting two correlated but separate processes related to nicotine dependence. In total, the results revealed that the FTND performs as well--from a psychometric perspective--with psychiatric smokers, as it does with nonpsychiatric smokers. PMID- 15893101 TI - Consumption of higher numbers of cigarettes in Mexican youth: the importance of social permissiveness of smoking. AB - This study sought to assess the relationship between level of cigarette smoking and social factors indicating permissiveness of smoking among youth. This was a school-based cross-sectional questionnaire study carried out in a random sample of students (n=13,293, 11-24 years) at public schools in Morelos, Mexico. Chi2 test and multiple logistic regression analysis were used to evaluate correlates of level of smoking (> or < or = 5 cigarettes/day). There is a rising trend in smoking among Mexican in-school youth, a tendency towards smoking initiation at increasingly younger ages and a decrease in the gender gap regarding smoking. When young people smoked at home, at school and outside, they were over fives times as likely to smoke >5 cigarettes/day. Prevention of smoking among Mexican adolescents should emphasize smoke-free schools, no smoking policies in the community and parental anti-tobacco education. PMID- 15893102 TI - Illicit drug use and abuse/dependence: modeling of two-stage variables using the CCC approach. AB - Drug use and abuse/dependence are stages of a complex drug habit. Most genetically informative models that are fit to twin data examine drug use and abuse/dependence independent of each other. This poses an interesting question: for a multistage process, how can we partition the factors influencing each stage specifically from the factors that are common to both stages? We used a causal common-contingent (CCC) model to partition the common and specific influences on drug use and abuse/dependence. Data on use and abuse/dependence of cannabis, cocaine, sedatives, stimulants and any illicit drug was obtained from male and female twin pairs. CCC models were tested individually for each sex and in a sex equal model. Our results suggest that there is evidence for additive genetic, shared environmental and unique environmental influences that are common to illicit drug use and abuse/dependence. Furthermore, we found substantial evidence for factors that were specific to abuse/dependence. Finally, sexes could be equated for all illicit drugs. The findings of this study emphasize the need for models that can partition the sources of individual differences into common and stage-specific influences. PMID- 15893103 TI - Under what circumstances are nurses willing to engage in brief alcohol interventions? A qualitative study from primary care in Sweden. AB - To improve alcohol prevention in primary health care, it has been suggested that primary care nurses are an under-utilised resource. The aim of this study was to identify under what circumstances primary care nurses in Sweden are willing to engage in alcohol prevention. All nurses at three primary health care centres in Ostergotland, Sweden were invited to participate in focus group interviews; 26 nurses participated. The nurses considered primary health care to be just one among other sectors within the community with responsibility for alcohol prevention. The role of health care in alcohol prevention was perceived to be important but mainly secondary preventive. The nurses felt justified screening all patients' alcohol habits only when they could refer to an obligation or a time-limited project. Otherwise, they mainly wanted to engage in screening patients with alcohol-related symptoms or diagnoses and other risk groups. Reasons for refraining from alcohol screening and intervention included lack of self-efficacy, time consumption and fear of harming their relationship with the patient. New strategies for alcohol prevention in primary care are discussed. PMID- 15893104 TI - Psychological distress and marijuana use before and after treatment: testing cognitive-behavioral matching hypotheses. AB - The purpose of this study was to examine the relationship between psychological distress, self-efficacy, and marijuana use using data from a randomized controlled trial of treatments for marijuana dependence [J. Consult. Clin. Psychol. 68 (2000) 898-908]. Adult marijuana users seeking treatment (N=291) were randomly assigned to three treatment conditions: (1) cognitive-behavioral relapse prevention support group (RPSG), (2) individualized assessment and advice group (IAI), and (3) delayed treatment control group (DTC). As predicted, psychologically distressed individuals had lower self-efficacy for avoiding marijuana use in psychologically distressing (PD) situations as opposed to nonpsychologically distressing (NPD) situations. However, all participants tended to have lower self-efficacy for NPD situations than PD situations. Efficacy increased and marijuana use decreased following treatment but the RPSG treatment did not have greater benefit for psychologically distressed participants. PMID- 15893105 TI - Childhood adversity and poor mothering: consequences of polydrug abuse use as a moderator. AB - Drug abuse consequences have been typically examined as a direct or main effect on various later outcomes. Drug abuse may also serve as a consequence that alters (moderates) critical developmental trajectories. This study examined the relationship between childhood adversity factors (parent alcohol and drug-related problems, childhood maltreatment) and future parenting practices through an analysis of the moderating effects of polydrug problems. Data from a community sample of mothers was divided into two groups based on the median split level of polydrug problems (alcohol, marijuana, cocaine) that they reported (low, N=123; high, N=114). Confirmatory factor models (CFAs) were developed for the two groups and compared with multiple group analyses (MGAs). Results revealed that mothers who had parents with alcohol or drug-related problems were more likely to become poor parents, if they themselves used drugs and had problems related to drug use. PMID- 15893106 TI - Predictors of attrition from day treatment of adolescents with substance-related disorders. AB - The study assessed the predictor variables of attrition from a day treatment for adolescents with substance-related disorders. The adolescents who were living in housing services and had more delinquent behaviors presented more difficulties to adhere, to attend regularly, and to maintain a link with the day program. The presence of cocaine among the substance-related disorders predicted an irregular attendance. Conversely, the adolescents with more self-reported symptoms of anxious and depression revealed a better initial adherence, a more regular attendance, and a longer length of the treatment. PMID- 15893107 TI - Impulsivity, negative expectancies, and marijuana use: a test of the acquired preparedness model. AB - According to the 'acquired preparedness model,' expectancies mediate the relationship between an impulsive personality style and alcohol use. The current study evaluated whether the model can also be applied to marijuana use. Estimated probabilities and subjective evaluations of personally expected marijuana effects, along with impulsivity and frequency of marijuana use, were assessed in 337 college undergraduates. Tests of mediation examining positive and negative marijuana expectancies showed negative expectancies to be a significant mediator for both males and females. That is, participants who were higher on impulsivity had fewer negative expectancies and in turn used more marijuana. This study provides evidence that the acquired preparedness model may help to explain marijuana use. PMID- 15893108 TI - Personality profiles of normal healthy research volunteers: a potential concern for clinical drug trial investigators? AB - "Normal healthy research volunteers" (NHRVs) in Phase I clinical trials are utilized to provide precise pharmacokinetic information and subjective self reports to help determine the initial drug dose for future Phase II, III, and IV trials, and accurate dose levels for future use in a patient population. Medical researchers rely on both objective pharmacokinetic and subjective data from NHRVs. Research shows that personality characteristics can influence these results in Phase I clinical trials. For example, extroverted personality characteristics have influenced the subjective reports and physiological experience of NHRVs. Second, subjective responses to placebos have been significantly greater in extroverted NHRVs than in introverted NHRVs. Third, in looking at Type-A and Type-B personality characteristics, NHRVs with Type-A Characteristics reported significantly more negative side effects to placebos than did people with Type-B characteristics. Fourth, NHRVs with high anxiety had more reports of adverse symptoms after drug administration than NHRVs with low anxiety. Further, it has been found that gastric emptying occurs more rapidly in people with high levels of neuroticism as compared to people with low levels of neuroticism. The authors suggest that personality and psychological characteristics of NHRVs can influence pharmacokinetic results and thus need to be addressed either in the statistical analysis or in the screening process for NHRV studies. Suggestions for further research and analysis are included, along with ideas for a national volunteer subject pool for NHRVs. PMID- 15893109 TI - The sweet thing about Type 1 diabetes: a cryoprotective evolutionary adaptation. AB - The reasons for the uneven worldwide distribution of Type 1 diabetes mellitus have yet to be fully explained. Epidemiological studies have shown a higher prevalence of Type 1 diabetes in northern Europe, particularly in Scandinavian countries, and Sardinia. Recent animal research has uncovered the importance of the generation of elevated levels of glucose, glycerol and other sugar derivatives as a physiological means for cold adaptation. High concentrations of these substances depress the freezing point of body fluids and prevent the formation of ice crystals in cells through supercooling, thus acting as a cryoprotectant or antifreeze for vital organs as well as in their muscle tissue. In this paper, we hypothesize that factors predisposing to elevated levels of glucose, glycerol and other sugar derivatives may have been selected for, in part, as adaptive measures in exceedingly cold climates. This cryoprotective adaptation would have protected ancestral northern Europeans from the effects of suddenly increasingly colder climates, such as those believed to have arisen around 14,000 years ago and culminating in the Younger Dryas. When life expectancy was short, factors predisposing to Type 1 diabetes provided a survival advantage. However, deleterious consequences of this condition have become significant only in more modern times, as life expectancy has increased, thus outweighing their protective value. Examples of evolutionary adaptations conferring selection advantages against human pathogens that result in deleterious effects have been previously reported as epidemic pathogenic selection (EPS). Such proposed examples include the cystic fibrosis mutations in the CFTR gene bestowing resistance to Salmonella typhi and hemochromatosis mutations conferring protection against iron-seeking intracellular pathogens. This paper is one of the first accounts of a metabolic disorder providing a selection advantage not against a pathogenic stressor alone, but rather against a climatic change. We thus believe that the concept of EPS should now include environmental factors that may be nonorganismal in nature. In so doing we propose that factors resulting in Type 1 diabetes be considered a result of environmental pathogenic selection (EnPS). PMID- 15893110 TI - Forecasting versus projection models in epidemiology: the case of the SARS epidemics. AB - In this work we propose a simple mathematical model for the analysis of the impact of control measures against an emerging infection, namely, the severe acute respiratory syndrome (SARS). The model provides a testable hypothesis by considering a dynamical equation for the contact parameter, which drops exponentially with time, simulating control measures. We discuss the role of modelling in public health and we analyse the distinction between forecasting and projection models as assessing tools for the estimation of the impact of intervention strategies. The model is applied to the communities of Hong Kong and Toronto (Canada) and it mimics those epidemics with fairly good accuracy. The estimated values for the basic reproduction number, R0, were 1.2 for Hong Kong and 1.32 for Toronto (Canada). The model projects that, in the absence of control, the final number of cases would be 320,000 in Hong Kong and 36,900 in Toronto (Canada). In contrast, with control measures, which reduce the contact rate to about 25% of its initial value, the expected final number of cases is reduced to 1778 in Hong Kong and 226 in Toronto (Canada). Although SARS can be a devastating infection, early recognition, prompt isolation, and appropriate precaution measures, can be very effective to limit its spread. PMID- 15893111 TI - The scavenger cell hypothesis of apoptosis: apoptosis redefined as a process by which a cell in living tissue is destroyed by phagocytosis. AB - Current literature on the definition and description of apoptosis is very confusing and erratic, due to voluminous studies in recent decades using cell culture technique. Apoptosis has evolved as a programmed mechanism of cell demise to get rid of the cells that are no longer needed by the body. The most important reason for a creature to use this mechanism to kill cells is to avoid inflammatory response that causes tissue damage and ensuing scar formation, as seen in necrosis. To reach this aim, the dying cell communicates, at early stages of the dying process, with macrophages or its neighboring cells that have phagocytotic ability, coined collectively as scavenger cells herein. The dying cell is swiftly engulfed by a scavenger cell without leaking any noxious cellular components into the intercellular space to provoke an inflammatory response. Thus, apoptosis is a process involving at least one other cell type and is actually a mechanism occurring in live tissue. Most studies of apoptosis in recent decades neglect this fundamental point and use cell culture system with a single cell type in the medium, in which avoidance of inflammatory response and tissue damage is no longer a reason. In culture, the dying cell has no way to signal scavenger cells to engulf itself and thus needs to demobilize a series of special mechanisms, which have no need in live tissue, to complete the suicidal process and clearance of its own corpse. These "otherwise-no-need" mechanisms seem to involve activation of executor caspases by cytochrome c, and the activated caspases mediate late processes of apoptosis in vitro. However, because the late processes of apoptosis in vivo actually occur in a phagosome of scavenger cell, it may be phagosomal enzymes, but not executor caspases of the apoptotic cell origin, that are really involved in apoptosis. Therefore, I propose a "scavenger cell hypothesis of apoptosis" to redefine apoptosis as an in vivo mechanism of cell death, and suggest that programmed cell death in culture in a third cell demise mechanism besides necrosis and apoptosis that should be defined using other nomenclatures. PMID- 15893112 TI - Are vasoactive neuropeptide autoimmune fatigue-related disorders mediated via G protein-coupled receptors? AB - Vasoactive neuropeptides such as pituitary adenylate cyclase activating polypeptide (PACAP), calcitonin gene related peptide (CGRP) and vasoactive intestinal peptide (VIP) have been implicated in a number of fatigue-related conditions. Associations of these vasoactive neuropeptides with heat shock proteins (hsps) and cytosine-guanosine dinucleotide (CpG) DNA fragments in autoimmune phenomena have been postulated to interfere with receptor signal activation for adenylate cyclase and other vital cellular processes. However, a specific mechanism for receptor dysfunction has not been explored to date. G protein-coupled receptors (GPCRs) constitute a high proportion of biological receptor mechanisms and serve a wide range of substances including nucleosides, nucleotides, catecholamines, calcium, histamine, serotonin and prostaglandins. They are complex transmembrane hepta-helical serpentine structures with specific binding capabilities resulting in conformational changes that activate cognate cyclic GMP (G proteins). GPCRs adapt to certain stimuli through desensitisation and changes in phosphorylation and are subject to distortions of signalling processes. Hence, these vital signalling structures are susceptible to impairment of function through a range of mechanisms. One of their vital functions is signalling through adenylate cyclase, a vital step in cyclic AMP metabolism. This step involves ATP metabolism and therefore is a crucial mediator of cellular energy pathways. Some GPCRs act to inhibit adenylate cyclase (Gi proteins). Also vasoactive neuropeptides, such as PACAP display a number of receptor isotypes including null variants. Overexpression of Gi proteins and null variant receptors may account for major disruptions of signal transduction and ATP/cAMP metabolism. This paper examines the possible role of GPCR dysfunction in contributing to fatigue-related vasoactive neuropeptide autoimmune disorders which may include chronic fatigue syndrome (CFS), Gulf War syndrome (GWS) and even sudden infant death syndrome (SIDS). PMID- 15893113 TI - p53 Protein is activated by Pin1: and also by Cu-SOD prion-like enzyme. AB - Peptidyl-proline isomerase (Pin1) is able to trigger some conformationally important change in the p53 protein: there is notable protection by p53 (tumour suppressor protein) of human cells that prevents their entry into the carcinogenesis-committed routeway. Pin1 controls the ready (low energy change) equilibrium between the cis and trans distinctive folding configurations differentially at a proline residue: this amino acid residue in proteins is unique in bending sharply its peptide chain (to 90 degrees change): in the cis rather than trans orientation with respect to the peptide bond to residue X "upstream" linked as XCONHR. Moreover p53 protein can arrest a cell cycle progression (or trigger apoptosis) by acting as a transcription factor to nuclear DNA acting at p53 nuclear responsive element controlling a larger number of genes that produce proteins that stop cell growth or stimulate apoptosis, in stressed cells. Oxidative stress by reactive oxygen species (ROS) is carcinogenic but also stops cell growth and triggers apoptosis, Cu-SOD removes ROS (see figure). Could superoxide dismutase (Cu-SOD), therefore, provide the DNA-damage direct second route (first route is binding of Pin1) in DNA-damaged cells to p53 activation? The p53 protein that prevents carcinogenesis is activated by Pin1. In addition, this p53 tumour suppressor protein is activated by Cu-SOD. PMID- 15893114 TI - PPARalpha activators may be good candidates as antiaging agents. AB - Aging is associated with a metabolic decline characterized by the development of changes in fat distribution, obesity, and insulin resistance. Dysfunctional humoral and cell-mediated immune responses occur with age, and these aberrations have been implicated in the increased incidence of infectious diseases, hyporesponsiveness to vaccination, and the etiology of numerous chronic degenerative diseases. All these metabolic and immune alterations are associated with a variety of age-related diseases that subsequently result in increased mortality. Leptin can modulate many of the metabolic alterations characteristic of aging. Leptin resistance has been implicated in the pathogenesis of obesity related complications involving abnormalities of lipid metabolism that resemble those of old age. Increased plasma leptin levels with aging suggest resistance to leptin action and may explain why elderly subjects have abdominal obesity and insulin resistance. Leptin's failure may be considered for the metabolic decline seen with aging. Peroxisome proliferator-activated receptor (PPAR)-alpha, the transcription factor for the mitochondrial and peroxisomal enzymes of beta oxidation, and its target enzymes, are upregulated by hyperleptinemia. PPARalpha has been shown to mediate the action of the hypolipidemic drugs of the fibrate class on lipid and lipoprotein metabolism. PPARalpha activators furthermore improve glucose homeostasis and influence body weight and energy homeostasis. The administration of agents capable of activating the PPARalpha was found to restore the cellular redox balance, evidenced by a lowering of tissue lipid peroxidation, an elimination of constitutively active NF-kappaB, loss in spontaneous inflammatory cytokine production, and ailing in the aging immunity. PMID- 15893115 TI - Are treatment emergent suicidality and decreased response to antidepressants in younger patients due to bipolar disorder being misdiagnosed as unipolar depression? AB - While antidepressants have been demonstrated to be a safe and effective treatment for unipolar major depressive disorder (MDD) in adults, the use of antidepressants to treat children and adolescents is controversial. There is a paucity of evidence to suggest that antidepressants are effective when used to treat children and adolescents and some recent placebo-controlled evidence has suggested that antidepressant treatment increases suicidality in this population. MDD is a very broad construct, and includes many clinical subtypes. Bipolar Disorder (BD) has an earlier age of onset than MDD. The initial polarity of illness in bipolar disorder is frequently depression. Patients are more likely to first present for treatment during the depressive phase of the illness than during manic or hypomanic phases. It is probable that a substantial portion of depressed children and adolescents may not suffer from unipolar MDD but may have a bipolar spectrum disorder. There are few trials supporting the efficacy of antidepressants in bipolar disorder, and some evidence that they may induce rapid cycling, switching and mania. Antidepressant induced mania is often mixed, with admixtures of manic and depressive features. An increased suicide risk is a particular feature of mixed states, potentially explaining why suicidal ideation can emerge with antidepressant treatment. Antidepressants are unlikely to somehow act differently in children than they do in adults. A more plausible explanation is that incipient bipolar disorder is often not diagnosed early in children and adolescents and the differential effects of antidepressants in this group is a result of differing diagnostic casemixes. PMID- 15893116 TI - The paradox of HIV nef function: resolution of the contradictions. AB - The function of nef, an accessory protein of HIV, has been highly controversial. Careful studies by respected investigators have ascribed diametrically opposed functions to nef with some groups claiming nef increases replication and viral infectivity while others argue nef inhibits replication and reduces infectivity. Replicative homeostasis resolves this superficially irreconcilable paradox, and indicates both positions could be true, if the exact function of nef is dependent on whether the nef tested is wild-type or variant with respect to the polymerase. The basis of resolution of the enigma posed by nef function is fundamental to and understanding of viral pathobiology. PMID- 15893117 TI - Risk analysis of ectoparasites acting as vectors for chronic wasting disease. AB - Prion diseases are rare neurodegenerative diseases of humans and animals with a lethal evolution. Animal prion infections, such as chronic wasting disease (CWD) and scrapie (sheep) have shown a pattern of horizontal transmission. CWD is an endemic disease that has been affecting thousands of domestic and wild cervids in US for the last three decades. The mode of contamination is not known, although direct contact between infected and non-infected animals via saliva, urine and feces have been considered. Increasing spread of CWD has raised concerns about the potential transmission to humans and the conversion of human prion protein by CWD-associated prions has been demonstrated in laboratory experiments. Fly larvae exposed to brain infected material were able to readily transmit scrapie to hamsters. Prion rods were identified in both larvae and fly pupae. New lines of evidence confirmed that adult flies are also able to express prion proteins. The most prevalent species of myiasis in cattle, sheep and wild cervids (Hypoderma spp.) present a very different life cycle from human myiasis, with a long contact with neurologic structures, such as the spinal canal and epidural fat, that are potentially rich in prion rods. Considering the huge amount of fly larvae that affects each animal, it is important to discuss the possibility that these ectoparasites could theoretically act as reservoirs and vectors for CWD and other prion diseases. It is critical to recognize all the possible factors involved in CWD transmission since ectoparasites could be handled in an easier way than the environmental persistence of infectious prions. PMID- 15893118 TI - Dysfunction of the anterior hippocampus: the cause of fundamental schizophrenic symptoms? AB - Recent evidence suggests there is an association between schizophrenia and dysfunction of the anterior hippocampus. Accordingly, this paper endeavors to show how the fundamental schizophrenic symptoms described by Bleuler might arise from deficiencies in normal hippocampal function. This effort is based on the idea that the hippocampus normally constructs a composite picture or worldview of the environment when conditions are novel or uncertain. Then when conditions become familiar, it influences emotion and behavior to be consistent with that worldview. The anterior hippocampus maintains the emotional component of a worldview through its interaction with the amygdala and hypothalamus, and supports executive frontal lobe activity by way of its output to the nucleus accumbens. From this perspective, the split between affect and cognition in schizophrenia is attributed to a failure of the anterior hippocampus to enforce the emotional component of a worldview allowing emotions to diverge from the cognitive component. The splitting of associations is traced to a failure of anterior hippocampal output to support frontal lobe control over the flow of associations. And a split from reality is seen to arise from a combination of these two failures. The individual ceases to interact with the world because the lack of executive control makes such interaction difficult. Fear and fantasies come to dominate experience because emotions are not adequately controlled. PMID- 15893119 TI - RISUG (reversible inhibition of sperm under guidance)--an antimicrobial as male vas deferens implant for HIV free semen. AB - HIV transmission from the male to the female is a major health problem. A hypothesis proposing an intra vas deferens implant of an antimicrobial compound to prevent the infection spread is presented. Mechanisms of action for the inhibition could include inactivating HIV in sperms passing through the vas deferens; drug release from the implant to destroy HIV entering into semen from genital structures distal to the vas deferens; and sperm acrosome released hyaluronidase mediated reabsorption of HIV. A subcomponent of the implant flowing along sperm pathway may have a role in reducing the entry of HIV from a positive female into penile tissue. A new drug RISUG (reversible inhibition of sperm under guidance) presently undergoing clinical trials for its contraceptive effect in the male (because it disrupts the sperm acrosome by an electrical charge and pH lowering effects) has also antimicrobial action. The drug being a combination of styrene maleic anhydride (SMA) and dimethyl sulfoxide (DMSO) on being injected into the lumen of the vas deferens produces styrene maleic acid thereby lowering pH; induces electrochemical action leading to a stable electrical charge generation; releases mandelic acid; and induces acrosome reaction in sperms with consequent release of hyaluronidase and sperm inactivation. Moreover, one time administration into the lumen of the vas gives long term action. All these phenomena very well match with the needs for HIV clearance of semen and hence RISUG is here proposed as a possible candidate material for the HIV inhibiting vas deferens implant when delivered in below contraceptive threshold dosage. For experimental validation, after obtaining data on the semen HIV load under control conditions in the HIV positive males inducted into the study, 30 mg of SMA in 120 microl of DMSO (contraceptive dose being 60 mg SMA+120 microl DMSO) is to be injected into vasa deferens bilaterally. Thereafter at intervals of one month the viral load needs to be determined in semen obtained either by masturbation or in lubricant free condom at intercourse - the method of collection remaining the same throughout for a particular subject. A significant reduction in the semen viral load following RISUG administration will validate the hypothesis. Speculated reduced female to male HIV transmission is more difficult to test. Nonspecific indications will come from a population study of the incidence of RISUG treated men becoming HIV positive as compared to that in the general population. PMID- 15893120 TI - Using histamine (H1) antagonists, in particular atypical antipsychotics, to treat anemia of chronic disease via interleukin-6 suppression. AB - Anemia of chronic disease (ACD) is a condition of decreased red cell mass secondary to some other chronic inflammatory condition. In ACD total body iron stores are normal, though serum iron is typically low secondary to iron sequestration by macrophages, and often iron supplementation is not an effective treatment for ACD for the same reason. The pathogenesis of ACD had been poorly understood, but recently there has been important progress: upregulation of interleukin-6 (Il-6) secondary to the underlying chronic inflammatory disease upregulates expression of the protein hepcidin. Upregulation of hepcidin causes anemia by a number of mechanisms: decreased intestinal absorption of iron from the duodenum, increased sequestration of iron by macrophages. Thus, downregulation of Il-6 may represent a most important treatment avenue for ACD. Anti-Il-6 antibodies might be a way to lower Il-6 levels, but such antibodies besides being expensive would have to be given intravenously or intramuscularly, and such large immunogenic molecules may not be appropriate in patients already with a chronic inflammatory condition. Here, we note that an immediately available and potentially effective treatment for ACD is to decrease Il-6 levels by histamine (H1) receptor antagonism, given that histamine acting through the H1 receptor is known to be a potent positive regulator of Il-6. Among the classes of medications that are H1 antagonists we point out that atypical antipsychotic medications such as olanzapine and quetiapine are among the most potent H1 antagonists, and can have simple daily dosing schedules and thus may be particularly useful in ACD. PMID- 15893121 TI - Does clozapine work by blocking spikes and sparing bursts? AB - Clozapine works better and produces fewer side effects than other antipsychotics. Existing hypotheses fail to explain why. A new hypothesis, single spike suppression, supposes that psychotic symptoms are mediated by the single spikes of neurons at the D2 receptor. All antipsychotics block these spikes. Clozapine, according to the hypothesis, blocks these spikes but, unlike other antipsychotics, spares the spike bursts that mediate movement, cognition and affect. This study explores the mathematical feasibility of single spike suppression. Could an antipsychotic with the right receptor kinetics selectively block single spikes? Could this selectivity have clinical consequences? To develop the hypothesis, the author made a mathematical model of the receptor occupancy of a synapse, and performed five simulations, varying input data within the range established by research. The effects of hypothetical antipsychotics on single spikes and bursts were compared. The author confirmed that a drug with the right dissociation rate constant (k off) would dissociate slowly enough to block single spikes, but rapidly enough to spare longer bursts. If the hypothesis is correct, this spike-selective, burst-sparing drug would work at relatively low D2 occupancies, and cause minimal D2-related side effects. Single spike suppression may explain the superior properties of clozapine better than competing hypotheses. If so, it would provide a better model for a new generation of safe, effective antipsychotics. PMID- 15893122 TI - Is autism caused by early hyperactivity of brain-derived neurotrophic factor? AB - Autism, a childhood-onset neurodevelopmental disorder, is characterized by disturbances in socialization and language skills, as well as in perception. Several studies indicate the importance of both genetic and environmental factors in the development of idiopathic autism, but the underlying pathogenesis of this disorder is still unknown. Brain-derived neurotrophic factor (BDNF) is important for normal neuronal development. Early BDNF hyperactivity may play an etiological role in autism early in life. This hypothesis is supported by the finding that serum and brain tissue BDNF levels are increased in autism compared with normal controls. Furthermore, BDNF hyperactivity may be associated with early brain outgrowth, increased prevalence of seizures in autism, and similar behaviors observed in autism and fragile X syndrome. Further studies of serum BDNF levels and genetic studies of the BDNF signaling pathway may help to clarify the role of BDNF in the pathogenesis of autism. Attempts to prove the BDNF hyperactivity hypothesis may lead investigators in a new direction for the study of the pathogenesis of autism and the development of an effective intervention of this disorder. PMID- 15893123 TI - The role of vestibular system and the cerebellum in adapting to gravitoinertial, spatial orientation and postural challenges of REM sleep. AB - The underlying reasons for, and mechanisms of rapid eye movement (REM) sleep events remain a mystery. The mystery has arisen from interpreting REM sleep events as occurring in 'isolation' from the world at large, and phylogenetically ancient brain areas using 'primal' gravity-dependent coordinates, reflexes and stimuli parameters to relay and process information about self and environment. This paper views REM sleep as a phylogenetically older form of wakefulness, wherein the brain uses a gravitoinertial-centred reference frame and an internal self-object model to evaluate and integrate inputs from several sensory systems and to adapt to spatial-temporal disintegration and malignant cholinergic-induced vasodepressor/ventilatory threat. The integration of vestibular and non vestibular sensory graviceptor signals enables estimation and control of centre of the body mass, position and spatial relationship of body parts, gaze, head and whole-body tilt, spatial orientation and autonomic functions relative to gravity. The vestibulocerebellum and vermis, via vestibular and fastigial nucleus, coordinate inputs and outputs from several sensory systems and modulate the amplitude and duration of 'fight-or-flight' vestibulo-orienting and autonomic 'burst' responses to overcome the ongoing challenges. Resolving multisystem conflicts during the unique stresses (gravitoinertial, hypoxic, thermal, immobilisation, etc.) of REM sleep enables learning, cross-modal plasticity, higher-order integration and multidimensional spatial updating of sensory-motor cognitive components. This paper aims to generate discussion, reinterpretation and creative testing of this novel hypothesis, which, if experimentally confirmed, has major implications across medicine, bioscience and space physiology, from developmental, clinical, research and theoretical perspectives. PMID- 15893124 TI - Unifying mechanism for addiction and toxicity of abused drugs with application to dopamine and glutamate mediators: electron transfer and reactive oxygen species. AB - There are many unknown aspects concerning the mode of action of abused drugs. Recently, a unifying theme for toxicity and addiction was reported based on electron transfer (ET), reactive oxygen species (ROS), and oxidative stress (OS). The main drugs involved are nicotine, cocaine, alcohol, phencyclidine, ecstasy, amphetamines, morphine-heroin, tetrahydrocannabinol, and therapeutic drugs (benzodiazepines, phenytoin, phenobarbital, aspirin, and acetaminophen). A major source of ROS is ET functionalities, of which the main ones found in abused drug metabolites are quinones and imines (or iminiums). Minor types are the nitroxide metabolite from cocaine, and alpha-dicarbonyl from alcohol. The theoretical approach enjoys support from reports on formation of ET metabolites, generation of ROS, protection by antioxidants (AOs), electrochemical studies, and cell signaling. Dopamine (DA) mediation of drug abuse has been the focus of much attention during the past decades. Recently, a similar role for glutamate (Glu) has come under study. Superficially, from a mechanistic vantage point, these findings might be regarded as in conflict with the ET-ROS-OS scheme. Many investigators believe that each drug or mediator operates by its own distinct mechanism. The present report provides evidence that a commonality in mode of action exists for both abused drugs and the DA-Glu operators. In the case of DA, oxidative metabolism yields o-quinones and semiquinones which can redox cycle with oxygen to provide various ROS. Electrochemical studies support the possibility of ET transformations by these quinones in the biological domain. In relation to cell signaling, DA is involved in formation of cAMP followed by a cascade of other events. A similar scenario exists in the case of Glu, in which an iminocarboxylic acid metabolite is hypothesized to play an ET role. The various phenomena are rationalized within the context of ET-ROS-OS, as was done earlier for abused drugs. Thus, a common mode of action may exist for both categories since all provide metabolites with similar properties. Stimulation of mediator production by abused drugs can occur with subsequent oxidation by ROS, some of which may be supplied by the drugs. In addition to prevention, the difficult topic of addiction mechanism is addressed from the viewpoint of ET and ROS involvement. Low levels of ROS would pertain since high concentrations are related to toxicity. A report that tetrahydrocannabinol does not comprise a substantial risk is in accord with little evidence of OS and absence of ET functionalities in the drug and reported metabolites. PMID- 15893125 TI - Will corticosteroids and other anti-inflammatory agents be effective for diarrhea predominant irritable bowel syndrome? AB - Irritable bowel syndrome (IBS) is one of several functional gastrointestinal disorders commonly encountered in both the clinical setting and the general population. The biopsychosocial model is currently believed to be a more complete explanatory mechanism of IBS symptom genesis and propagation. Gut inflammation and immune activation is one of the biological mechanisms for which evidence is emerging. Experimental parasitic infection of mice bowel resulted in elevated substance P levels and increased expression of cyclooxygenase 2 (COX 2) enzyme, prostaglandin E2, IL-4, IL-5, and IL-13. In IBS patients, increased cellularity and proximity of the inflammatory or immune cells to the nerve trunks of the bowel, elevated interleukin-1beta mRNA expression in mucosal biopsies, and increased inducible nitric oxide synthase and nitrotyrosine elaboration (indicative of lymphocyte activation) were observed. Corticosteroids given after the elimination of an experimentally applied parasite from the bowel of mice resulted in the reversal of persistent gut muscle dysfunction. Selective COX-2 inhibitors attenuated the increased bowel smooth muscle contractility resulting from parasite infection of mice gut. In humans, it has been observed that the relative risk of developing IBS in asthma patients was reduced by 60% by the use of oral steroids. Despite such preclinical and human evidence for the role of inflammation and immune activation in IBS, the efficacy of anti-inflammatory and immunomodulatory agents has not been adequately investigated. Budesonide, a corticosteroid with a high mucosal activity and a low bioavailability, is an anti inflammatory agent that may be worth investigating for its utility in diarrhea predominant IBS. PMID- 15893126 TI - Enhancing anti-inflammatory cytokine IL-10 may be beneficial for acute coronary syndrome. AB - Inflammation is one of the basic biological responses against vascular injury. It has been demonstrated, however, that inflammation is an important pathogenic feature in atherosclerotic lesions. Previous data showed that pro-inflammatory cytokines played an important role in acute coronary events. A complex intravascular inflammatory response is also an integral component of atherosclerotic plaque dynamic instability, and the imbalance between anti inflammatory mechanisms and pro-inflammatory factors, in favor of the pro inflammatory factors, will result in rupture of atherosclerotic plaque. Interluekine-10 (IL-10), which is produced by various inflammatory cells, especially macrophages, has multifaceted anti-inflammatory properties, including inhibition of the prototypic pro-inflammatory transcription factor nuclear factor kappaB, leading to suppressed cytokine production, inhibition of matrix degrading metalloproteinase production, reduction of tissue factor expression, inhibition of apoptosis of macrophages and monocytes after infection, and promotion of the phenotypic switch of lymphocytes into the Th2 phenotype. All these inflammatory mechanisms have been shown to play a pivotal role for atheroslerotic lesion development and progression, indicating a potential regulatory role of IL-10. More recent data showed that decreased plasma concentration of anti-inflammatory cytokine IL-10 was associated with acute coronary syndrome. Therefore, we hypotheses that enhancing anti-inflammatory cytokine IL-10 may be a promising approach for the therapy of acute coronary diseases. PMID- 15893127 TI - Dominant and opportunistic leukemic clones: proposal for a pathogenesis-oriented classification in acute myeloid leukemia. AB - Despite the common clinical, hematological and prognostic features that define acute myeloid leukemia (AML) there is considerable heterogeneity among individual cases, suggesting different pathogenic pathways. Based on a simple theoretical model, according to the vital characteristics of the leukemic clone (proliferative rate and resistance to apoptosis), I propose a classification of AML into two broad categories: (a) high leukemic clone vitality (HLV) AML or "dominant type" AML, corresponding roughly to the World Health Organization (WHO) classification group of entities "AML with recurrent cytogenetic abnormalities" and (b) low leukemic clone vitality (LLV) or "opportunistic type" AML corresponding to the WHO groups "AML with multilineage dysplasia" and "alkylating agent-related AML". HLV-AML leukemic clones are characterized by rate-limiting genomic mutations capable of conferring proliferation/survival advantage over a normal hematopoietic environment while in LLV-AML, the leukemic clones are not particularly proliferative or apoptosis-resistant, but are nevertheless selected against an impaired, previously damaged hematopoietic environment. Such a pathogenesis-oriented classification might have therapeutic and prognostic implications, providing a theoretical basis for a further adaptation of the current standard treatment strategies to the individual characteristics of the AML patients. PMID- 15893128 TI - Migraine prophylactic drugs work via ion channels. AB - In recent decades, the concept of a vascular origin of migraine has been replaced by theories based on a neuronal pathophysiology. These theories all involve rapid changes in the functioning of the brain, particularly the brain stem, and the trigeminal nerves. While such paroxysmal changes in function could be the result of altered synaptic transmission, or other physiological changes, they could also be due to changes in the function of voltage-regulated sodium and calcium ion channels. Support for this view of migraine as a channelopathy comes from an examination of the likely mechanism of action of migraine prophylactic drugs. It is the present hypothesis that most of the widely used drugs for migraine prevention work by inhibiting the function of one or both of these ion channels. A review of the laboratory research done on most of the commonly used migraine prophylactic drugs, divided into five classes, reveals that they all may work on sodium channels, calcium channels, or both. If this is the common mechanism of action of migraine prophylactics, it should lead toward the development of more effective prophylactic drugs. PMID- 15893129 TI - Alzheimer's disease and the 'ABSENT' hypothesis: mechanism for amyloid beta endothelial and neuronal toxicity. AB - Alzheimer's disease [AD] is the most common cause of dementia among people age 65 and older. One of the biggest stumbling blocks in developing effective drug therapy for Alzheimer's disease has been the lack of a comprehensive hypothesis that explains the mechanism behind all of the histopathological changes seen in patients suffering from Alzheimer's disease. An overview of the currently popular 'amyloid' and 'vascular' hypotheses for AD demonstrates that neither hypothesis by itself can explain all the known histopathological and biochemical lesions seen in Alzheimer's disease. The paper presents a hypothesis that tries to explain the mechanism behind almost all the histopathological changes, and varying clinical manifestations seen in both diagnosed AD and Vascular Dementia [VaD]. The new hypothesis is based on the known dual toxicity of beta amyloid to both vascular and neuronal tissues, their synergy and the resultant net effect on the onset and progression of AD. The new hypothesis therefore will be known as the Amyloid Beta Synergistic Endothelial and Neuronal Toxicity [ABSENT] hypothesis. The ABSENT hypothesis will try to show the common chemical mechanism behind almost all of the pathological changes seen in AD. According to the ABSENT hypothesis, beta amyloid itself generates all the free radicals that cause both vascular dysfunction and the neuronal damage seen in AD. The chemical mechanism proposed is based on evidence from physical chemistry experiments, calculations as well as in vitro/in vivo experiments. The ABSENT hypothesis does not favor one mode of beta amyloid-induced brain damage over the other, rather it considers the net effects of the neuronal stress/damage caused by both the cerebrovascular dysfunction and direct neurotoxicity caused by beta amyloid. The hypothesis states that each patient has a different balance of predisposing factors that modulate the extent of neurotoxicity and cerebrovascular dysfunction caused by beta amyloid and thereby explains the wide range and mixed nature of damage and dysfunction seen in the studies done on patients diagnosed with AD, VaD or 'mixed dementias'. According to the hypothesis, beta amyloid peptides are necessary if not sufficient to cause AD, VaD and mixed senile dementias. The hypothesis, therefore, proposes the term Beta Amyloid Dementias [BAD] to describe the conditions currently covered by the diagnoses of 'AD', 'VaD' and 'Mixed [senile] Dementias'. Finally, the ABSENT hypothesis tries to put forth a direct chemical mechanism behind the apparent synergy and increased association between old age, pre- and coexisting vascular disease, diabetes and AD. PMID- 15893130 TI - 5-Hydroxytryptophan plus SSRIs for interferon-induced depression: synergistic mechanisms for normalizing synaptic serotonin. AB - Interferon-alpha (IFN) is widely used in the treatment of certain cancers and viral infections, including hepatitis C (HCV). Unfortunately, depression is a common side effect of IFN therapy, affecting approximately a third of HCV patients receiving IFN therapy. Studies have shown that selective serotonin reuptake inhibitors (SSRIs) can effectively treat IFN-induced depression in only 63-75% of cases. For the remaining percentage, depression often necessitates dose reduction of or discontinuation from IFN therapy. Emerging evidence indicates that IFN may cause depression by affecting brain serotonin. IFN has been shown to increase serotonin reuptake and to decrease serotonin synthesis. We hypothesize that SSRIs are not fully effective because they affect only serotonin reuptake, not serotonin synthesis, and that effective treatment must address both uptake and synthesis. 5-Hydroxytryptophan (5-HTP) effectively increases central nervous system synthesis of serotonin. It is the immediate precursor of serotonin and is widely available as a dietary supplement, which is well absorbed after an oral dose. Several double-blind studies have shown 5-HTP to be effective in the treatment of nondrug-induced depression. We hypothesize that patients who become depressed on IFN will respond to the synergistic combination of SSRIs plus 5-HTP. PMID- 15893131 TI - Are cardiac syndrome X, irritable bowel syndrome and reflex sympathetic dystrophy examples of lateral medullary ischaemic syndromes? AB - Altered pain appreciation and autonomic function are hallmarks of Cardiac syndrome X, Irritable bowel syndrome and Reflex sympathetic dystrophy. Both pain appreciation and autonomic function are controlled by the lateral medulla. This hypothesis proposes that lateral medullary ischaemia at a microvascular level is responsible for these syndromes and could also be linked to other conditions where autonomic dysfunction is a major feature such as late-onset asthma, type 2 diabetes and essential hypertension. Autonomic function is controlled by the nucleus tractus solitarius, which acts as the main viscero-afferent nucleus in the brain stem regulating vagal tone. It is particularly susceptible to ischaemia since it is highly metabolically active and lies in a medullary arterial watershed zone. The anatomical route of the vertebral artery through cervical vertebra makes it vulnerable to injury from whiplash with or without any genetic predisposition to atheroma formation. This could make microvascular occlusion commonplace and a plausible explanation for the above syndromes. Ischaemia rather than infarction occurs because of the excellent collateral blood supply in the brainstem. In support of this hypothesis, a new Transcranial doppler ultrasonography arterial signal has been described called small vessel knock, the ultrasound signal of small vessel occlusion. Recent evidence has shown that ultrasound targeting of this signal in the vertebral artery improves clinical symptoms in these syndromes which supports this hypothesis. Two such cases are discussed. PMID- 15893132 TI - Concept-matching in the brain depends on serotonin and gamma-frequency shifts. AB - A Eureka moment has three components--puzzle, solution and hedonic response (elation etc.). Puzzle and solution come together in the association cortex and are immensely variable from instance to instance. By contrast, the hedonic response is subcortical and almost one-dimensional; how is it triggered? It is triggered by the relation between puzzle and solution, a good fit or good match, like the relation between two words that rhyme. In 1999 J.W. Fost proposed that serotonin is a crucial agent; here it is proposed that a frequency-jump initiates the serotonin causal chain, as energy shifts from 20 to 40 Hz or some such jump. The hypothesis assumes that any discrete idea is embodied in a time-course of electrical and chemical changes in a network of neurons, and that keeping the idea in mind involves repeating more or less the same time-course over and over. If observed frequencies in the gamma range result from such repetition, the period for running the time-course once is of the order of 25 ms. Also accepted is the suggestion that, although the brain runs many processes simultaneously, in the conscious mind attention focuses on only one idea at a time; an attempt to "think of two things at once" actually results only in giving them attention alternately, with a repeat-time of the order of 50 ms and frequency 20 Hz. Only if the two time-courses have certain elements in common will there be any repetition at 40 Hz. Now suppose a thinker takes up a problem and makes a succession of attempts at solution. As long as he thinks of wrong answers, he generates activity only at 20 Hz, but when he hits upon the right answer, activity at 40 Hz shows up. This is a highly oversimplified scenario but its essential features might carry over to the vastly more complicated workings of a real brain. The virtue of the proposed mechanism is its generality. Under the proposal, any ideas in mind that do not match give no result but as soon as two ideas match, results ensue. This behavior in the model, wholly general except in one specific respect, is needed for conformity with real human brains' behavior. In normal people, production of this "link-joy" is an important reward mechanism and malfunction of this system may contribute to Capgras syndrome and some varieties of autism. PMID- 15893133 TI - Inhibition of postprandial hyperglycemia by acarbose is a promising therapeutic strategy for the treatment of patients with the metabolic syndrome. AB - The metabolic syndrome is strongly associated with insulin resistance and has been recognized as a cluster of risk factors for cardiovascular diseases such as visceral obesity, hypertension, diabetes, and atherogenic dyslipidemia. Recently, insulin resistance in the absence of overt diabetes or the metabolic syndrome itself has been associated with endothelial dysfunction, one of the initial steps in the process of atherosclerosis. Postprandial hyperglycemia, one of the characteristic features of insulin resistance, induces oxidative stress generation and elicits vascular inflammation and platelet activation, thus being involved in the pathogenesis of atherosclerosis. A recent multicenter, placebo controlled randomized trial, STOP-NIDDM trial, revealed that acarbose (Glucobay R), an alpha-glucosidase inhibitor, improved postprandial hyperglycemia and subsequently reduced the risk of development of type 2 diabetes in patients with impaired glucose tolerance (IGT). In this study, acarbose treatment was also found to slow the progression of intima-media thickness of the carotid arteries, a surrogate marker for atherosclerosis, and to reduce the incidence of cardiovascular diseases and newly diagnosed hypertension in subjects with IGT. Acarbose significantly reduced body mass index and waist circumference in these patients over 3 years. Furthermore, a meta-analysis of seven long-term studies has also shown that intervention with acarbose prevents myocardial infarction and cardiovascular diseases in type 2 diabetic patients. In this analysis, glycemic control, triglyceride levels, body weight and systolic blood pressure was also significantly improved during acarbose treatment. These observations suggest that prevention of postprandial hyperglycemia by acarbose may be a promising therapeutic strategy for reducing the increased risk for diabetes, hypertension, dyslipidemia, obesity, and cardiovascular diseases in patients with the metabolic syndrome. Acarbose improves postprandial hyperglycemia by delaying the release of glucose from complex carbohydrates in the absence of an increase in insulin secretion. Therefore, we would like to hypothesize here that this improvement in glucose metabolism could be associated with amelioration in insulin sensitivity, thus explaining the above-mentioned beneficial cardiometabolic actions of acarbose. Large clinical trials will provide us with more definite information whether acarbose treatment can improve insulin sensitivity and resultantly reduce the risk of diabetes, hypertension and cardiovascular diseases in patients with the metabolic syndrome. PMID- 15893134 TI - Unique atheroprotective property of azelnidipine, a dihydropyridine-based calcium antagonist. AB - Insulin resistance and central obesity are often associated with hypertension. The metabolic syndrome is a cluster of these common clinical disorders, and is related with an increased risk for cardiovascular diseases. A number of pro inflammatory cytokines derived from adipose tissues have been thought to contribute to the development of insulin resistance and accelerated atherosclerosis. Among them, TNF-alpha has been most widely studied; it not only suppresses the insulin signaling, but also elicits vascular inflammation. Indeed, inhibition of TNF-alpha was found to improve insulin resistance in obese rats and reduce the progression of atherosclerosis in apolipoprotein E knockout mice, respectively. These observations demonstrate that TNF-alpha could play a central role in the pathogenesis of insulin resistance and accelerated atherosclerosis in the metabolic syndrome. Considering that the primary goals of treatment for hypertensive patients with the metabolic syndrome are prevention of the development of diabetes and cardiovascular events, anti-hypertensive drugs that have abilities to block the TNF-alpha signaling would be desirable as a first line therapy for these patients. In the process of the search for such a unique anti-hypertensive drug, we have recently found that azelnidipine, a newly developed and commercially used long-acting dihydropyridine-based calcium antagonist (DHP), inhibited TNF-alpha-induced activator protein-1 activation and interleukin-8 expression in human umbilical vein endothelial cells by suppressing NADPH oxidase-mediated reactive oxygen species generation. The concentration of azelnidipine that was found effective in these in vitro-experiments is well within the therapeutic range. Since endothelial cells do not possess voltage operated L-type calcium channels, these observations suggest that the beneficial effects of azelnidipine are not likely due to calcium channel blocking property, but due to its unique anti-oxidative ability. Furthermore, we have very recently found that serum levels of monocyte chemoattractant protein-1, a biomarker for subclinical atherosclerosis, were significantly decreased by the treatment of azelnidipine in patients with essential hypertension. In this paper, we would like to hypothesize that due to its unique TNF-alpha signal modulatory, anti oxidative property, azelnidipine may be a promising DHP that targets diabetes and cardiovascular diseases in hypertensive patients with the metabolic syndrome. PMID- 15893135 TI - Proposed multigenic Composite Inheritance in major depression. AB - Various rationale have been considered in the familial inheritance pattern of major depression ranging from simple one-gene Mendelian inheritance to pseudo additive gene action. We instead predict broad genetic expressivity patterns in the progeny of parents where at least one parent has recurrent major depression. In keeping with this idea, we feel that recurrent major depression could involve an expression imbalance of "normal" genes either exclusively or along with allelic variation(s). The patterns of pathology are theoretically conceptualized as qualitative and quantitative, meaning that expressivity of the genetic pattern in these children may range from minimal to complete even among siblings. Thus, prediction of the particular genetic pattern expressed by a particular child might prove difficult. The complex inheritance pattern that we propose is referred to as Composite Inheritance. Composite Inheritance considers that both the up- and down-regulation of luxury genes and housekeeping genes are involved in this dichotomous qualitative inheritance pattern and also the wide quantitative expressivity. The luxury genes include such genes as those coding for the neurotransmitter transporters and receptors. The housekeeping genes found to date include those that code for proteins involved in gene transcription, secondary signaling systems, fatty acid metabolism and transport, and intracellular calcium homeostasis. Other luxury and housekeeping genes no doubt remain to be discovered. Our current research utilizes an empirical approach involving advanced genomics and specialized pattern recognition mathematics in families having at least one parent with recurrent major depression. The goal of our research is to develop a pattern recognition system of genetic expressivity in major depression to which prevention and early intervention may be tailored. PMID- 15893136 TI - The dynamic range of biologic functions and variation of many environmental cues may be declining in the modern age: implications for diseases and therapeutics. AB - We hypothesize that declining dynamic range and variation of environmental cues may contribute to health dysfunctions, and that judicious expansion of biologic dynamic ranges may be beneficial. Three disparate examples involving the endocrine, autonomic, and musculoskeletal systems are discussed. Daytime sheltering, optical shading, and nighttime use of artificial light may reduce circadian luminal variation. The resulting melatonin alterations may contribute to systemic dysfunctions. Loss of temporal variation of other hormones may contribute to biologic dysfunctions, especially those involving the hypothalamic pituitary axis. Reduced variation of physical exertion, environmental stressors, and thermal gradients that characterize modern lifestyles may reduce the autonomic dynamic range resulting in lowered heart rate variability and a myriad of systemic dysfunctions. The health benefits of activities such as exercise, meditation, acupuncture, coitus, and laughter may operate through increasing autonomic variability. Reduced physical exertion also accounts for declining dynamic range of musculoskeletal function. The resulting muscle atrophy, fat infiltration, and sarcomere shortening may not only have deleterious local effects, but may also be involved in systemic metabolic dysfunctions such as insulin resistance. The extent to which our endogenous systems rely on environmental variation for self-tuning and the impact that under-utilization of compensatory mechanisms has on biologic function are not well understood. Modern therapeutic approaches generally result in reversion to the mean of physiologic functions and may buffer against variation. For example, beta-blockers are given to reduce adrenergic excess, insulin to treat insulin insufficiency, serotonin reuptake inhibitors for depression, and refractive lenses for myopia. By undermining the demand for native compensatory functions, such therapeutic strategies may actually impair future ability to respond to biologic disequilibria. Generalizing from these observations, we anticipate benefits of therapeutic and lifestyle approaches that expand, rather than reduce, the dynamic range of many biologic experiences. PMID- 15893137 TI - Male reproductive physiology as a sexually selected handicap? Erectile dysfunction is correlated with general health and health prognosis and may have evolved as a marker of poor phenotypic quality. AB - Many extravagant physical traits are selected because they are used as cues for mate choice (sexual selection). Why is mate choice driven by costly ornaments? A theory of sexual selection posits that extravagant traits are preferred because are reliable indicators of superior (heritable) phenotypic quality. In particular, the preferred traits can be expressed only in individuals with superior conditions because are handicaps which impose a high cost to the carrier. The human penis achieves its reproductive function by the complex neuro vascular mechanisms that controls erection. Surprisingly, erectile dysfunction and infertility, two condition which nearly annihilate fitness, are widespread medical conditions which affect millions of people of any age worldwide. The very high incidence of erectile dysfunction appears as an evolutionary paradox. Impotence is associated with all major systemic diseases as well depression and stress. Stress is also one of the causes of infertility. Therefore, male reproduction appears to be extremely sensitive to internal and external stressors. Moreover, erectile dysfunction is a predictor of myocardial infarction and stroke, whereas men with regular sexual activity have lower risk of death due to coronary disease. This large body of medical literature makes erection the best case for a fitness indicator in the human male. We suggest that the fragility of male sexual physiology observed in modern society is the specific consequence of an evolutionary process driven by the handicap principle. PMID- 15893138 TI - Kaiser Wilhelm syndrome: obstetric trauma or placental insult in a historical case mimicking Erb's palsy. AB - Kaiser Wilhelm II of Germany (1859) developed a weak and noticeably short left arm during childhood, commonly attributed to nerve damage caused by the use of excessive force during his difficult breech delivery, Erb's palsy. However, Wilhelm's mother had a severe fall when about four months pregnant and the child was reported to be very thin at birth, suggesting intrauterine growth restriction (IUGR). Wilhelm blamed the British doctor for his deformity, and formed an enmity, which ultimately led to the 1914-1918 world war. We propose an alternative theory, considering the possibility of placental damage and consequential flow redistribution caused by the fall. In severe IUGR, the Doppler pulsatility index (PI) of the brachial arteries differs, that of the right arm being lower than the left. We used a computer model of the fetoplacental unit and reduced its functional placental area until such resistance asymmetry was produced. This would occur in extreme hypoxia when flow in the aortic isthmus is reversed, bringing right ventricular blood of lower oxygen content to the left subclavian artery. The reduced PI in the right arm is a normal vasodilatory hypoxic response, but the apparently normal PI in the left arm results from decreased demand due to metabolic failure. We suggest that the nerve damage affecting the Kaiser's left arm was due to placental insufficiency during pregnancy, and not mechanical brachial plexus injury during delivery. We further suggest that such a mechanism be called Kaiser Wilhelm syndrome to distinguish it from Erb's palsy originating from obstetric trauma. PMID- 15893139 TI - Should we use digit preference as an indicator of quality control? PMID- 15893140 TI - On the similarity of cocaine and 3,3'-iminodipropionitrile. PMID- 15893141 TI - Pure and mixed hormone receptor positive breast tumors: are they different from therapeutic point of view? PMID- 15893142 TI - How to understand autistic motivational status. PMID- 15893143 TI - Atavistic orofacial response to manually dextrous activities. PMID- 15893146 TI - A cybernetic theory of aging. PMID- 15893145 TI - Neutrophil chemotaxis: autotaxin induced attenuation of purine/pyrimidine signaling precede the overexpression of Integrin(s). PMID- 15893147 TI - Nanotechnology helps medicine: nanoscale swimmers and their future applications. PMID- 15893148 TI - Making cell-permeable recombinant telomerase (trans-telomerase) through fusion of its catalytic subunit (hTERT) with protein transduction domains (PTD): a possible strategy to overcome replicative senescence during ex vivo culture of primary explanted cells. PMID- 15893149 TI - Anti-inflammatory effects of long-chain omega 3 fatty acids: potential benefits for atrial fibrillation. PMID- 15893150 TI - Transcendental meditation does not predispose to epilepsy. PMID- 15893151 TI - EEG, hypometabolism, and ketosis during transcendental meditation indicate it does not increase epilepsy risk. PMID- 15893152 TI - The contribution of low plasma taurine to clinical complications of Homocystinurea. PMID- 15893154 TI - Self-rated everyday and prospective memory abilities of cigarette smokers and non smokers: a web-based study. AB - The present study examined self-ratings of two aspects of everyday memory performance: long-term prospective memory-measured by the prospective memory questionnaire (PMQ), and everyday memory-measured by the everyday memory questionnaire (EMQ). Use of other substances was also measured and used as covariates in the study. To ensure confidentiality and to expand the numbers used in previous studies, an Internet study was carried out and data from 763 participants was gathered. After controlling for other drug use and strategy use, the data from the PMQ revealed that smokers reported a greater number of long term prospective memory errors than non-smokers. There were also differences between light and heavier smokers in long-term prospective memory, suggesting that nicotine may have a dose-dependent impact upon long-term prospective memory performance. There was also a significant ANOVA group effect on the EMQ, although the trend for more memory errors amongst the heavier smokers was statistically only borderline (p=.057). These findings suggest there are selective memory deficits associated with smoking and that long-term prospective memory deficits should be added to the growing list of problems associated with cigarette use. PMID- 15893155 TI - Profiling the subjective, psychomotor, and physiological effects of a hydrocodone/acetaminophen product in recreational drug users. AB - BACKGROUND: Hydrocodone (HYD) is a mu opioid agonist. Hydrocodone/acetaminophen HYD/ACET compounds are both widely prescribed and abused prescription painkillers in the United States. In the present study, we profiled the subjective, psychomotor, and physiological effects of the compound. It was of particular interest to determine if HYD/ACET had abuse liability-related subjective effects in a population of recreational drug users. METHODS: Eighteen volunteers participated in a crossover, randomized, double-blind study in which they received, all p.o., placebo; 5 mg HYD/500 mg ACET; 10mg HYD/500 mg ACET; 20 mg HYD/1000 mg ACET; 40 mg morphine (MOR) sulfate; and 1000 mg ACET. Measures were assessed before and for 300 min after drug administration. RESULTS: HYD/ACET produced effects that were dose-related, and the highest dose produced a similar magnitude of effect to that of MOR. There were some abuse liability-related subjective effects produced by 20 mg HYD/ACET and MOR, but there were also unpleasant effects. Some unpleasant subjective effects were experienced only by females. Overall liking and "take again" ratings assessed 24 h post-session were not significant, but several subjects had elevated liking and "take again" ratings at this time in one or more of the HYD/ACET conditions and/or in the MOR condition. Both 20 mg HYD/1000 mg ACET and MOR impaired psychomotor performance. HYD/ACET and MOR produced miosis. CONCLUSIONS: HYD/ACET produced some abuse liability-related subjective effects in recreational drug users, which is consistent with the widespread non-medical use and abuse of this product. PMID- 15893156 TI - Early onset inhalant use and risk for opiate initiation by young adulthood. AB - In this study, we estimate a hypothesized link from early onset inhalant use to later use of opiates by young adulthood, with data from an epidemiological sample of 2311 first graders who entered an urban mid-Atlantic public school system in 1985 or 1986 (49.8% male; 67.1% ethnic minority), and who were studied longitudinally to young adulthood. An estimated 9% had initiated inhalant use before the age of 14 and at follow-up in young adulthood an estimated 3% (n=66) of the sample had tried opiates at least once. Youth who used inhalants prior to age 14 were twice as likely to initiate opiate use, as compared to those who had never tried (relative risk 2.2; 95% CI=1.4, 3.3). Statistical adjustment for other covariates attenuated but did not dissolve this relationship. These findings help confirm previously reported evidence that the use of inhalants might be an early marker of vulnerability for future involvement with illegal drugs such as heroin, but an exploratory analysis suggests that there may be no direct inhalants--opiate link once a general early onset susceptibility trait is taken into account. PMID- 15893157 TI - Temporal dynamics and determinants of whole brain tissue volume changes during recovery from alcohol dependence. AB - Brain shrinkage and its partial reversibility with abstinence is a common neuroimaging finding in alcohol dependent individuals. We used an automated three dimensional whole brain magnetic resonance imaging method (boundary shift integral) in 23 alcohol dependent individuals to measure the temporal dynamics of cerebral tissue and spinal fluid volume changes over a 12-month interval and to examine the major determinants of brain tissue change rates during abstinence and non-abstinence. We found more rapid brain tissue gain during the first month of sobriety than in the following months. The most rapid volume recovery was observed in abstinent individuals with the greatest baseline brain shrinkage and drinking severity. The rapid reversal of brain volume gains in non-abstinent individuals and tissue volume changes are modulated by duration of abstinence and non-abstinence periods, as well as recency of non-abstinence. Age, family history density of alcoholism, relapse severity, and duration or age of onset of heavy drinking were not major determinants of brain shrinkage and brain volume recovery rates. Treatment providers may use this tangible information to reinforce the biomedical benefits of sobriety. Previous quantitative measurements of brain volumes in alcohol dependent individuals performed after several weeks of abstinence likely underestimated the full extent of chronic alcohol-associated brain shrinkage. PMID- 15893158 TI - Do research payments precipitate drug use or coerce participation? AB - Providing high-magnitude cash incentives to substance abuse clients to participate in research is frequently viewed as unethical based on the concerns that this might precipitate new drug use or be perceived as coercive. We randomly assigned consenting drug abuse outpatients to receive payments of 10 US dollars, 40 US dollars, or 70 US dollars in either cash or gift certificate for attending a 6-month research follow-up assessment. At the 6-month follow-up, participants received their randomly determined incentive and were then scheduled for a second follow-up appointment 3 days later to detect new instances of drug use. Findings indicated that neither the magnitude nor mode of the incentives had a significant effect on rates of new drug use or perceptions of coercion. Consistent with the contingency management literature, higher payments and cash payments were associated with increased follow-up rates. Finally, the results suggest that higher magnitude payments may be more cost-effective by reducing the need for more intensive follow-up efforts. PMID- 15893159 TI - Neuropsychological functioning in methadone maintenance patients versus abstinent heroin abusers. AB - Several studies have reported on neuropsychological status as an important contributing variable in drug abuse rehabilitation outcomes. However, few studies have dealt with cognitive impairment in methadone maintenance patients (MMP), despite the fact that methadone is the most frequently used opioid substitution treatment in European countries. The objective of the present study is to contrast the neuropsychological performance of MMP with that of abstinent heroin abusers (AHA). Participants were matched with respect to age, education, pre morbid IQ, employment status and lifetime drug abuse, and they underwent a set of tests aimed at assessing visuo-spatial attention, processing speed and executive functions. Although processing speed and attention deficits have previously been the focus of studies with MMP, executive functions have not received a similar degree of attention. The purpose of comparing matched MMP and AHA is two-fold: firstly, to test the differential effects of current opioid consumption and past opioid abuse on cognitive-executive performance and secondly, to assess the potential consequences of opioid-related neuropsychological deficits. Results showed a significantly slower performance by MMP on processing speed, visuo spatial attention, and cognitive flexibility tests (Five Digit Test (FDT) parts 1 and 3; Oral Trails (OT) parts 1, 2; Interference 2-1), and less accuracy in working memory and analogical reasoning tests extracted from the Wechsler Adult Intelligence Scale (WAIS III). Effect sizes for significant comparisons ranged from 0.67 to 1. These results seem to suggest that methadone consumption by itself induces significant cognitive impairments that could compromise drug treatment outcomes in MMP. PMID- 15893160 TI - Effect of selective antagonism of mu(1)-, mu(1/2)-, mu(3)-, and delta-opioid receptors on the locomotor-stimulating actions of ethanol. AB - Previous studies have demonstrated that administration of nonspecific opioid antagonists such as naltrexone or naloxone reduces ethanol-induced locomotor activity in mice. However, because of their broad pharmacological profile, it remains unclear through which opioid receptor this antagonism is achieved. Therefore, the present study was aimed at further investigating the role of the different opioid receptors in ethanol-induced (2.5 g/kg) locomotion in mice. First, we compared the effect of naltrexone (0-2 mg/kg) on ethanol-induced locomotion with that of the selective delta-opioid receptor antagonist, naltrindole (0-10 mg/kg). Results of this first set of data revealed that naltrexone completely blocked this effect of ethanol at doses suggested to occupy only mu-opioid receptors, and naltrindole did not modify ethanol-induced locomotion. In a second set of experiments, we further investigated the involvement of mu-opioid receptors in ethanol-stimulated motor activity by assessing the implication of mu(1)-, mu(1/2)-, and mu(3)-opioid receptor subtypes. Results revealed that mu(1/2)-, and to a lesser extent mu(3)-, but not mu(1)-opioid receptor subtypes are involved in the psychomotor actions of ethanol. Data are discussed together with previous results which have emphasized the critical dependence of ethanol-induced motor behaviors on opioid receptors, as well as, of the integrity of beta-endorphin synthesizing neurons from the hypothalamic Arcuate Nucleus. PMID- 15893161 TI - Drinking refusal self-efficacy questionnaire-revised (DRSEQ-R): a new factor structure with confirmatory factor analysis. AB - The drinking refusal self-efficacy questionnaire (DRSEQ) assesses a person's belief in their ability to resist alcohol. The DRSEQ is a sound psychometric instrument based on exploratory factor analyses, but has not been subjected to confirmatory factor analysis. In total 2773 participants were used to confirm the factor structure of the DRSEQ. Initial analyses revealed that the original structure was not confirmed in the current study. Subsequent analyses resulted in a revised factor structure (DRSEQ-R) being confirmed in community, student and clinical samples. The DRSEQ-R was also found to have good construct and concurrent validity. The factor structure of the DRSEQ-R is more stable than the original structure of the DRSEQ and the revised scale has considerable potential in future alcohol-related research. PMID- 15893162 TI - Depression among entrants to treatment for heroin dependence in the Australian Treatment Outcome Study (ATOS): prevalence, correlates and treatment seeking. AB - AIM: To determine the rate of current major depressive disorder (MDD) among entrants to treatment for heroin dependence in three treatment modalities and a non-treatment comparison group; and to ascertain factors associated with depression. DESIGN: Cross sectional structured interview. SETTING: Sydney, Australia. PARTICIPANTS: 615 current heroin users: 201 entering methadone/buprenorphine maintenance (MT), 201 entering detoxification (DTX), 133 entering drug free residential rehabilitation (RR) and 80 not in treatment (NT). FINDINGS: Current major depressive episode was reported by 25%. The rates of major depressive disorder ranged from 26% in the treatment groups (23% MT, 25% DTX, 31% RR) to 16% of those not in treatment. Females were more likely to have current major depressive episode (31% versus 21% OR 1.70, 95% CI 1.16-2.48). Factors associated with depression in the treatment groups were post traumatic stress disorder (PTSD), attempted suicide in the last 12 months and severe physical disability. Among the non-treatment group those with depression were also more likely to have PTSD. Women entering treatment were three times more likely to meet criteria for current major depression than women not in treatment. Among men however, the rates were not significantly different. CONCLUSION: Depression is a significant concern among entrants to treatment for heroin dependence. An essential component of treatment should be a consideration of depression, with the provision of appropriate treatment were required. PMID- 15893163 TI - Memory performance in polyvalent MDMA (ecstasy) users who continue or discontinue MDMA use. AB - BACKGROUND: The popular dance drug ecstasy (3,4 methylenedioxymethamphetamine=MDMA) is a serotonergic neurotoxin in animal studies. Several cross-sectional investigations reported low memory and learning performance in ecstasy users, particularly in those reporting heavy patterns of drug use. Since, serotonin has a recognized role in memory processes, these findings were mostly interpreted as evidence for ecstasy-related neurotoxicity in humans. However, studies with user populations and controls suffer from many inherent methodological problems. Moreover, longitudinal data on memory performance after continued or discontinued ecstasy use are scarce. METHODS: In the present longitudinal study, we examined memory performance in 38 MDMA users over the course of 18 months. RESULTS: Subjects who stopped MDMA use after the baseline examination (n=17) did not improve, and subjects who continued MDMA use (n=21) did not deteriorate in terms of test performance. CONCLUSIONS: Our data do not support, but they also do not rule out memory decline following use of the serotonergic neurotoxin MDMA. In light of the popularity of ecstasy among young people, further investigations are needed. In our view, research strategies should now move to prospective designs in order to shed more light on the course of possible adverse cognitive effects of ecstasy use. PMID- 15893164 TI - Utilizing Recovery Management Checkups to shorten the cycle of relapse, treatment reentry, and recovery. AB - Over the past several decades, a growing body of evidence suggests that a subset of substance users suffers from what appears to be a more chronic condition, whereby they cycle through periods of relapse, treatment reentry, incarceration, and recovery, often lasting several years. Using data from quarterly interviews conducted over a 2-year period in which 448 participants were randomly assigned to either an assessment only condition or to a Recovery Management Checkup (RMC) condition, we looked at the frequency, type, and predictors of transitions between points in the relapse, treatment reentry, and recovery cycle. The results indicated that about one-third of the participants transitioned from one point in the cycle to another each quarter; 82% transitioned at least once, 62% multiple times. People assigned to RMC were significantly more likely to return to treatment sooner and receive more treatment. The probability of transitioning to recovery was related to the severity, problem orientation, desire for help, self efficacy, self-help involvement, and recovery environment at the beginning of the quarter and the amount of treatment received during the quarter. These findings clearly support the wide spread belief that addiction is a chronic condition as well as demonstrating the need and effectiveness of post-discharge monitoring and checkups. The methods in this study also provide a simple but replicable method for learning more about the multiple pathways that individuals travel along before achieving a prolonged state of recovery. PMID- 15893165 TI - Wine preference and related health determinants in a U.S. national sample of young adults. AB - This study examined relationships between wine preference and selected health determinants in a U.S. national sample of young adults to improve understanding of the association between light-moderate wine consumption and long-term morbidity and mortality risk. Interview data collected from 12,958 young adults who participated in the National Longitudinal Study of Adolescent Health were analyzed to determine whether wine preference was related to educational, health and lifestyle characteristics that are predictive of long-term morbidity and mortality. Wine drinkers generally had more formal education, better dietary and exercise habits, and more favorable health status indicators (e.g., normal body mass) than other drinkers and non-drinkers. A larger proportion of wine drinkers were light-moderate drinkers compared to beer or liquor drinkers, and wine drinkers were less likely to report smoking or problem drinking than beer or liquor drinkers. These findings indicate that wine preference in young adulthood is related to educational, health and lifestyle characteristics that may help to explain the association between light-moderate wine consumption and morbidity, and mortality risk in later adulthood. PMID- 15893166 TI - Use of specialty substance abuse and mental health services in adults with substance use disorders in the community. AB - AIMS: To examine the patterns and correlates of use of specialty substance abuse and mental health services among adults with alcohol or non-alcohol drug abuse or dependence in the community. METHODS: Analyses focused on 5,568 participants with alcohol or non-alcohol drug abuse or dependence drawn from a large representative cross-sectional survey of the US general population-the 2002 US National Survey on Drug Use and Health (NSDUH). RESULTS: Only 9.7% of adults with substance use disorders used specialty substance abuse services in the past year; 22.4% used mental health services. Severity of substance use disorder and less education were associated with using substance abuse services. Whereas psychological distress and impairment in role functioning due to psychological problems were associated with mental health service use. Male gender, black race/ethnicity, and lack of health insurance acted as barriers to using mental health services but not specialty substance abuse services. Past year use of substance abuse services, but not mental health services, was associated with lower likelihood of continued use of substances in the past month. CONCLUSIONS: Individuals with substance use disorders are more likely to use mental health services than specialty substance abuse services. However, only people who use specialty substance abuse services have a lower risk of continued use of substances. Findings highlight the need for integration of substance abuse treatments in the mental health care system and attention to different barriers to the two types of services. PMID- 15893167 TI - The effects of physical activity on serum C-reactive protein and inflammatory markers: a systematic review. AB - Physical activity is associated with a reduced incidence of coronary disease, but the mechanisms mediating this effect are not defined. There has been considerable recent interest in inflammation in the pathogenesis of cardiovascular disease. Some of the beneficial role of physical activity may result from its effects on the inflammatory process. We searched PubMed for articles published between 1975 through May 2004 using the terms exercise, physical activity, or physical fitness combined with C-reactive protein, inflammation, inflammatory markers, or cytokines. The review revealed 19 articles on the acute inflammatory response to exercise, 18 on cross-sectional comparisons of subjects by activity levels, and 5 examining prospectively the effects of exercise training on the inflammatory process. Exercise produces a short-term, inflammatory response, whereas both cross-sectional comparisons and longitudinal exercise training studies demonstrate a long-term "anti-inflammatory" effect. This anti-inflammatory response may contribute to the beneficial effects of habitual physical activity. PMID- 15893168 TI - Multiple overlapping drug-eluting stents to treat diffuse disease of the left anterior descending coronary artery. AB - OBJECTIVES: We sought to determine the safety and efficacy of using multiple overlapping drug-eluting stents (DES) in patients with diffuse left anterior descending coronary artery (LAD) disease. BACKGROUND: Diffuse LAD disease represents a therapeutic challenge. Results after coronary artery bypass surgery are suboptimal, whereas the use of bare metal stents is limited by high rates of restenosis. The introduction of DES prompted treatment of long diffuse disease with multiple overlapping stents. METHODS: All consecutive patients with de novo diffuse LAD disease treated with more than 60-mm long DES from April 2002 to March 2004 were analyzed. RESULTS: The study population consisted of 66 patients. Thirty-nine patients were treated with sirolimus-eluting stents (SES), average length 84 +/- 22 mm, and 27 patients with paclitaxel-eluting stents (PES), average length 74 +/- 14 mm. The number of stents implanted per patient was 2.8 +/- 0.7, whereas the mean total stent length for the LAD treatment was 80 +/- 20 mm. Angiographic as well as procedural success was achieved in 95% of cases. Eleven (16.6%) patients had in-hospital non-Q-wave myocardial infarction (five SES and six PES), and one patient developed intraprocedural stent thrombosis. All patients had clinical follow-up, and 52 patients (79%) had an angiographic follow up at six months. Hierarchical major adverse cardiac event rate was 15% (7.5% for SES and 7.5% for PES). No patients died, one patient had non-Q-wave myocardial infarction (non-index vessel), and 10 patients (15%) underwent target vessel revascularization. CONCLUSIONS: The implantation of multiple overlapping DES in patients with a diffusely diseased LAD is relatively safe and associated with good midterm clinical outcomes. PMID- 15893169 TI - Endothelial progenitor cell capture by stents coated with antibody against CD34: the HEALING-FIM (Healthy Endothelial Accelerated Lining Inhibits Neointimal Growth-First In Man) Registry. AB - OBJECTIVES: This study was designed to evaluate whether rapid endothelialization of stainless steel stents with a functional endothelium prevents stent thrombosis and reduces the restenotic process. BACKGROUND: A "pro-healing" approach for prevention of post-stenting restenosis is theoretically favored over the use of cytotoxic or cytostatic local pharmacologic therapies. It is believed that the central role of the vascular endothelium is to maintain quiescence of the underlying media and adventitia. METHODS: Sixteen patients with de novo coronary artery disease were successfully treated with implantation of endothelial progenitor cell (EPC) capture stents. RESULTS: Complete procedural and angiographic success was achieved in all 16 patients. The nine-month composite major adverse cardiac and cerebrovascular events (MACCE) rate was 6.3% as a result of a symptom-driven target vessel revascularization in a single patient. There were no other MACCE despite only one month of clopidogrel treatment. At six month follow-up, mean angiographic late luminal loss was 0.63 +/- 0.52 mm, and percent stent volume obstruction by intravascular ultrasound analysis was 27.2 +/ 20.9%. CONCLUSIONS: This first human clinical investigation of this technology demonstrates that the EPC capture coronary stent is safe and feasible for the treatment of de novo coronary artery disease. Further developments in this technology are warranted to evaluate the efficacy of this device for the treatment of coronary artery disease. PMID- 15893170 TI - High-dose folic acid acutely improves coronary vasodilator function in patients with coronary artery disease. AB - OBJECTIVES: We investigated the acute effect of orally administered high-dose folic acid on coronary dilator function in humans. BACKGROUND: Folic acid and its active metabolite, 5-methyltetrahydrofolate, increase endothelium-dependent vasodilation in human peripheral circulation. However, the acute effect on coronary circulation is not known. METHODS: Fourteen patients with ischemic heart disease, age 62 +/- 12 years (mean +/- SD), were enrolled in a double-blind, placebo-controlled crossover trial. Basal and adenosine-stimulated myocardial blood flow (MBF) were determined by positron emission tomography, and myocardial flow reserve was calculated. Each patient was studied after ingestion of placebo and after ingestion of 30 mg folic acid. Myocardial zones were prospectively defined physiologically as "normal" versus "abnormal" on the basis of MBF response to adenosine 140 microg/kg/min (normal = MBF >1.65 ml/min/g). Abnormal and normal zones were analyzed separately in a patient-based analysis. RESULTS: Folate was associated with a reduction in mean arterial pressure (100 +/- 12 mm Hg vs. 96 +/- 11 mm Hg, placebo vs. folate, p < 0.03). Despite the fall in mean arterial pressure, folic acid significantly increased the MBF dose response to adenosine (p < 0.001 using analysis of variance) in abnormal zones, whereas MBF in normal zones did not change. In abnormal segments, folic acid increased peak MBF by 49% (1.45 +/- 0.59 ml/min/g vs. 2.16 +/- 1.01 ml/min/g, p < 0.02). Furthermore, folate increased dilator reserve by 83% in abnormal segments (0.77 +/- 0.59 vs. ml/min/g 1.41 +/- 1.08 ml/min/g, placebo vs. folate, p < 0.05), whereas dilator reserve in normal segments remained unchanged (2.00 +/- 0.61 ml/min/g vs. 2.12 +/- 0.69 ml/min/g, placebo vs. folate, p = NS). CONCLUSIONS: The data demonstrate that high-dose oral folate acutely lowers blood pressure and enhances coronary dilation in patients with coronary artery disease. PMID- 15893171 TI - Diffuse and active inflammation occurs in both vulnerable and stable plaques of the entire coronary tree: a histopathologic study of patients dying of acute myocardial infarction. AB - OBJECTIVES: This study was undertaken to define and compare geographic coronary artery inflammation in patients who were dying of acute myocardial infarction (AMI), chronic stable angina (SA), and noncardiac causes (CTRL). BACKGROUND: Biochemical markers and flow cytometry provide indirect evidence of diffuse coronary inflammation in patients dying of acute coronary syndromes. Yet no histopathologic studies have corroborated these findings. A key unanswered question is whether the inflammatory burden involves the entire coronary tree or is limited to a few plaques. METHODS: We examined 544 coronary artery segments from 16 patients with AMI, 109 segments from 5 patients with SA, and 304 coronary segments from 9 patients with CTRL. RESULTS: An average of 6.8 +/- 0.5 vulnerable segments per patient were found in the AMI group (in addition to culprit lesions) compared with an average of 0.8 +/- 0.3 and 1.4 +/- 0.3 vulnerable lesions/patient in the SA and CTRL groups, respectively. The AMI group, independent of the type of plaque observed, showed significantly more inflammatory infiltrates compared with the SA and CTRL groups (121.6 +/- 12.4 cell x mm2 vs. 37.3 +/- 11.9 cell x mm2 vs. 26.6 +/- 6.8 cell x mm2, p = 0.0001). In AMI patients, active inflammation was not only evident within the culprit lesion and vulnerable plaques but also involved stable plaques. These showed a three- to four-fold higher inflammation than vulnerable and stable plaques from the SA and CTRL groups, respectively. CONCLUSIONS: This histopathologic study found that both vulnerable and stable coronary plaques of patients dying of AMI are diffusely infiltrated by inflammatory cells. PMID- 15893172 TI - Multiple plaque rupture and C-reactive protein in acute myocardial infarction. AB - OBJECTIVES: This study sought to investigate the relationship between multiple plaque ruptures, C-reactive protein (CRP), and clinical prognosis in acute myocardial infarction (AMI). BACKGROUND: Several studies have demonstrated that ruptured or vulnerable plaques exist not only at the culprit lesion but also in the whole coronary artery in some acute coronary syndrome (ACS) patients. Recent studies have reported that a ruptured plaque at the culprit lesion is associated with elevated CRP, which indicates a poor prognosis in patients with ACS. METHODS: We performed intravascular ultrasound in 45 infarct-related arteries and another 84 major coronary arteries in 45 first AMI patients. RESULTS: Plaque rupture was observed in 21 patients (47%) at the culprit site. Intravascular ultrasound revealed 17 additional plaque ruptures at remote sites in 11 patients (24%). Patients with multiple risk factors were more frequently found in our multiple-plaque rupture patients compared with single-plaque rupture or nonrupture patients (82% vs. 40% vs. 29%, p = 0.01). High-sensitive CRP levels had a positive correlation with the number of plaque ruptures (p < 0.01). All culprit lesions were successfully treated by percutaneous coronary intervention. Patients with multiple plaque rupture showed significantly poor prognosis compared with others (p = 0.01). CONCLUSIONS: Multiple plaque rupture is associated with systemic inflammation, and patients with multiple plaque rupture can be expected to show a poor prognosis. Our results suggest that AMI treatment should focus not only on stabilization of the culprit site but also a systemic approach to systemic stabilization of the arteries. PMID- 15893173 TI - Act local, act global: inflammation and the multiplicity of "vulnerable" coronary plaques. PMID- 15893174 TI - Increased carotid atherosclerosis in andropausal middle-aged men. AB - OBJECTIVES: This study examined the association between carotid artery intima media thickness (IMT), serum sex hormone levels, and andropausal symptoms in middle-aged men. BACKGROUND: Male sex hormones may play a dual role in the pathogenesis of atherosclerosis in men by carrying both proatherogenic and atheroprotective effects. METHODS: We studied 239 40- to 70-year-old men (mean +/ SD: 57 +/- 8 years) who participated in the Turku Aging Male Study and underwent serum lipid and sex hormone measurements. Ninety-nine men (age 58 +/- 7 years) were considered andropausal (i.e., serum testosterone <9.8 nmol/l or luteinizing hormone [LH] >6.0 U/l and testosterone in the normal range), and in both situations, they had subjective symptoms of andropause (a high symptom score in questionnaire). Three were excluded because of diabetes. The rest of the men (age 57 +/- 8 years) served as controls. Carotid IMT was determined using high resolution B-mode ultrasound, and serum testosterone, estradiol (E2), LH, and sex hormone-binding globulin were measured using standard immunoassays. RESULTS: Andropausal men had a higher maximal IMT compared with controls in the common carotid (1.08 +/- 0.34 vs. 1.00 +/- 0.23, p < 0.05) and in the carotid bulb (1.44 +/- 0.48 vs. 1.27 +/- 0.35, p = 0.003). Common carotid IMT correlated inversely with serum testosterone (p = 0.003) and directly with LH (p = 0.006) in multivariate models adjusted for age, total cholesterol, body mass index, blood pressure, and smoking. CONCLUSIONS: Middle-aged men with symptoms of andropause, together with absolute or compensated (as reflected by high normal to elevated LH) testosterone deficiency, show increased carotid IMT. These data suggest that normal testosterone levels may offer protection against the development of atherosclerosis in middle-aged men. PMID- 15893175 TI - Andropause and intima media thickness. PMID- 15893177 TI - What promise does PCSK9 hold? PMID- 15893176 TI - A common PCSK9 haplotype, encompassing the E670G coding single nucleotide polymorphism, is a novel genetic marker for plasma low-density lipoprotein cholesterol levels and severity of coronary atherosclerosis. AB - OBJECTIVES: We sought to determine the effects of PCSK9 variants on plasma low density lipoprotein cholesterol (LDL-C) levels, severity of coronary atherosclerosis, and response to statin therapy in the Lipoprotein Coronary Atherosclerosis Study (LCAS) population. BACKGROUND: Mutations in PCSK9 cause autosomal-dominant hypercholesterolemia. We hypothesized that PCSK9 variants could affect plasma LDL-C in individuals with polygenic hypercholesterolemia. METHODS: We sequenced all 12 exons and boundaries to detect novel polymorphisms, and genotyped 372 subjects in LCAS and 319 subjects in a second independent population for six polymorphisms, including novel leucine repeats, by fluorescently tagged markers. We reconstructed haplotypes using a Bayesian algorithm. RESULTS: Permutation test results showed statistically significant differences in global haplotype distribution among the tertiles of LDL-C (odds ratio [OR]: 2.36, 95% confidence interval [CI]: 1.90 to 4.32, p = 0.005) and minimum lumen diameter of coronary lesions (OR: 1.83, 95% CI: 1.01 to 3.55, p = 0.045). Regression analysis identified haplotype 3 as an independent determinant of LDL-C levels (adjusted R2 = 2.2%, F = 9.37, p = 0.002). Haplotype structure analysis identified E670G as the determinant variant, exerting a dose effect (GG > EG > EE) and accounting for 3.5% of plasma LDL-C variability (F = 14.6, p < 0.001). Plasma total cholesterol, apolipoprotein B, and lipoprotein (a) levels were also associated with the E670G variant. Distributions of the E670G genotypes in an independent normolipidemic and the hyperlipidemic LCAS populations were significantly different (F = 7.2, p = 0.027). No significant treatment-by genotype interactions were detected. The false positive report probability was between 2% and 8%. CONCLUSIONS: Haplotype 3 encompassing the E670G variant is an independent determinant of plasma LDL-C levels and the severity of coronary atherosclerosis. PMID- 15893178 TI - Elevated levels of VE-cadherin-positive endothelial microparticles in patients with type 2 diabetes mellitus and coronary artery disease. AB - OBJECTIVES: The purpose of this study was to examine whether CD144-EMP (endothelium-derived microparticles) is useful as a specific marker of endothelial cell (EC) dysfunction and to determine whether plasma levels of circulating CD144-EMP predicted coronary artery disease (CAD) in patients with type 2 diabetes mellitus (DM). BACKGROUND: Endothelial cell dysfunction is involved in atherogenesis; however, the quantitative assessment of EC dysfunction has yet to be established clinically. Endothelium-derived microparticles are small, membrane-shed vesicles that are generated from the EC surface in response to cellular dysfunction and/or injury. Diabetes mellitus is known to be associated with EC dysfunction and accelerated atherosclerosis. METHODS: We characterized EMP using anti-CD144 (VE-Cadherin) antibody in various atherosclerosis-related cells and investigated the association between the levels of CD144-positive microparticles and hydrogen-peroxide-induced EC injury and acetylcholine-induced coronary vasomotion. Furthermore, we evaluated plasma CD144 EMP levels in patients with and without DM. RESULTS: We demonstrated that CD144 positive microparticles were derived selectively from human EC. The levels of CD144-EMP reflected the degree of in vitro hydrogen-peroxide-induced EC injury and impairment of in vivo endothelium-dependent coronary vasodilation (p < 0.01). Plasma CD144-EMP levels were increased significantly in DM patients compared with patients without DM (p < 0.001). In DM patients, the elevated levels of CD144-EMP were the most significant risk factor for CAD relative to all other traditional risk factors (odds ratio [OR] 3.5, 95% confidence interval [CI] 1.8 to 6.9, p < 0.001). Notably, plasma CD144-EMP identified a subpopulation of established CAD patients in DM subjects without typical anginal symptoms (OR 10.6, 95% CI 3.9 to 29.5, p < 0.001). CONCLUSIONS: The CD144-positive EMP exist in human plasma, and plasma CD144-EMP levels can be a clinically specific and quantitative marker of EC dysfunction and/or injury. Measurement of CD144-EMP, by providing a quantitative assessment of EC dysfunction, may be useful for identifying DM patients with increased risk of CAD. PMID- 15893179 TI - Glutathione peroxidase-1 and homocysteine for cardiovascular risk prediction: results from the AtheroGene study. AB - OBJECTIVES: This prospective study was designed to evaluate the effect of joint determination of two important contrary biomarkers--homocysteine and glutathione peroxidase (GPx)-1--on cardiovascular risk stratification. BACKGROUND: Homocysteine plasma levels have been associated with cardiovascular risk. Experimental data suggest that antioxidative GPx-1 activity modulates cardiovascular risk associated with homocysteine. METHODS: In 643 patients with coronary artery disease, we performed a prospective study to assess the risk of homocysteine plasma levels and GPx-1 activity on long-term cardiovascular risk with a median follow-up of 7.1 years. RESULTS: Both homocysteine and GPx-1 were among the strongest univariate predictors of future cardiovascular risk, even after adjustment for cardiovascular confounders. Homocysteine levels were significantly elevated in individuals with future cardiovascular events (15.4 vs. 13.4 micromol/l; p < 0.0001); GPx-1 activity was lower (45.3 +/- 13.1 vs. 50.2 +/ 11.0 U/g hemoglobin; p < 0.0001). In patients with GPx-1 activity below the median value, homocysteine plasma levels above the median were associated with a 3.2-fold (95% confidence interval 1.8 to 5.6; p < 0.0001) increase in cardiovascular risk, whereas it lost its independent risk prediction in individuals with increased antioxidative capacity, as reflected by high GPx-1 activity. In contrast to single determination, combined assessment revealed a significant increase in the area under the curve of cardiovascular risk predictive models from 0.72, including traditional risk factors to 0.75 and also including homocysteine levels and GPx-1 activity. CONCLUSIONS: Plasma homocysteine levels and GPx-1 activity are complementary in identifying individuals at high cardiovascular risk. Joint determination of both biomarkers provides substantial information on top of classic risk factors in cardiovascular risk assessment. PMID- 15893180 TI - Which white blood cell subtypes predict increased cardiovascular risk? AB - OBJECTIVES: We sought to determine the predictive ability of total white blood cell (WBC) count and its subtypes for risk of death or myocardial infarction (MI). BACKGROUND: An elevated WBC count has been associated with cardiovascular risk, but which leukocyte subtypes carry this risk is uncertain. METHODS: Consecutive patients without acute MI who were assessed angiographically for coronary artery disease (CAD) and were followed up long-term were studied. The predictive ability for death/MI of quartile (Q) 4 versus Q1 total WBC, neutrophil (N), lymphocyte (L), and monocyte (M) counts and N/L ratio were assessed using Cox regressions. RESULTS: A total of 3,227 patients was studied. Mean age was 63 years; 63% of patients were male, and 65% had CAD. In multivariable modeling entering standard risk factors, presentation, and CAD severity, the total WBC (hazard ratio [HR] 1.4, p = 0.01) and M (HR 1.3, p < 0.02) were weaker and N (HR 1.8, p < 0.001), L (HR 0.51, p < 0.001), and N/L ratio (HR 2.2, p < 0.001) were independent predictors of death/MI. When WBC variables were entered together, N/L ratio and M were retained as independent predictors. Risk associations persisted in analyses restricted to CAD patients or including acute MI patients. CONCLUSIONS: Total WBC count is confirmed to be an independent predictor of death/MI in patients with or at high risk for CAD, but greater predictive ability is provided by high N (Q4 >6.6 x 10(3)/microl) or low L counts. The greatest risk prediction is given by the N/L ratio, with Q4 versus Q1 (>4.71 versus <1.96) increasing the hazard 2.2-fold. These findings have important implications for CAD risk assessment. PMID- 15893181 TI - Relative efficacy of atorvastatin 80 mg and pravastatin 40 mg in achieving the dual goals of low-density lipoprotein cholesterol <70 mg/dl and C-reactive protein <2 mg/l: an analysis of the PROVE-IT TIMI-22 trial. AB - OBJECTIVES: The aim of this research was to compare relative efficacy of different statin regimens in achieving the dual goals of low-density lipoprotein cholesterol (LDL-C) and C-reactive protein (CRP) reduction. BACKGROUND: While secondary prevention guidelines for statin therapy suggest lowering LDL-C levels <70 mg/dl, we have recently shown that clinical outcomes are improved when CRP levels are also lowered <2 mg/l. METHODS: We addressed the relative efficacy of pravastatin 40 mg and atorvastatin 80 mg daily to reduce LDL-C and CRP among 3,745 acute coronary syndrome patients. RESULTS: A total of 1,018 participants (27.1%) achieved the dual goals of LDL-C <70 mg/dl and CRP <2 mg/l. After adjustment for age, gender, smoking, diabetes, hypertension, obesity, and HDL-C, these individuals had a 28% lower risk of recurrent myocardial infarction or vascular death (relative risk = 0.72; 95% confidence interval 0.52 to 0.99). Of those who achieved dual goals, 80.6% received atorvastatin 80 mg, while 19.4% received pravastatin 40 mg (p < 0.001). Only 11% allocated pravastatin and 44% allocated atorvastatin achieved the goals of LDL-C <70 mg/dl and CRP <2 mg/l, and only 5.8% allocated pravastatin 40 mg and 26.1% allocated atorvastatin 80 mg reached the even lower goals of LDL-C <70 mg/dl and CRP <1 mg/l. The correlation coefficient for CRP measured at 30 days and at end of study was 0.61 (p < 0.001), a value almost identical to that for LDL-C over the same follow-up period (r = 0.62, p < 0.001). CONCLUSIONS: While atorvastatin 80 mg was superior to pravastatin 40 mg in terms of achieving the dual goals of aggressive LDL-C and CRP reduction, neither agent brought the majority of patients below thresholds needed to maximize patient benefit. PMID- 15893182 TI - Additive beneficial effects of fenofibrate combined with atorvastatin in the treatment of combined hyperlipidemia. AB - OBJECTIVES: We compared vascular and metabolic responses (and adverse responses) to statin and fibrate therapies alone or in combination in patients with combined hyperlipidemia. BACKGROUND: The mechanisms of action for statins and fibrates are distinct. METHODS: Fifty-six patients were given atorvastatin 10 mg and placebo, atorvastatin 10 mg and fenofibrate 200 mg, or fenofibrate 200 mg and placebo daily during each two-month treatment period of a randomized, double-blind, placebo-controlled crossover trial with two washout periods of two months' each. RESULTS: Lipoproteins were changed to a greater extent with combined therapy when compared with atorvastatin or fenofibrate alone. Flow-mediated dilator response to hyperemia and plasma high-sensitivity C-reactive protein and fibrinogen levels were changed to a greater extent with combined therapy when compared with atorvastatin or fenofibrate alone (p < 0.001, p = 0.182, and p = 0.015 by analysis of variance [ANOVA], respectively). The effects of combined therapy or fenofibrate alone on plasma adiponectin levels and insulin sensitivity (determined by the Quantitative Insulin-Sensitivity Check Index [QUICKI]) were significantly greater than those of atorvastatin alone (p = 0.022 for adiponectin and p = 0.049 for QUICKI by ANOVA). No patients were withdrawn from the study as the result of serious adverse effects. CONCLUSIONS: Combination therapy is safe and has beneficial additive effects on endothelial function in patients with combined hyperlipidemia. PMID- 15893183 TI - Noninvasive home telemonitoring for patients with heart failure at high risk of recurrent admission and death: the Trans-European Network-Home-Care Management System (TEN-HMS) study. AB - OBJECTIVES: We sought to identify whether home telemonitoring (HTM) improves outcomes compared with nurse telephone support (NTS) and usual care (UC) for patients with heart failure who are at high risk of hospitalization or death. BACKGROUND: Heart failure is associated with a high rate of hospitalization and poor prognosis. Telemonitoring could help implement and maintain effective therapy and detect worsening heart failure and its cause promptly to prevent medical crises. METHODS: Patients with a recent admission for heart failure and left ventricular ejection fraction (LVEF) <40% were assigned randomly to HTM, NTS, or UC in a 2:2:1 ratio. HTM consisted of twice-daily patient self measurement of weight, blood pressure, heart rate, and rhythm with automated devices linked to a cardiology center. The NTS consisted of specialist nurses who were available to patients by telephone. Primary care physicians delivered UC. The primary end point was days dead or hospitalized with NTS versus HTM at 240 days. RESULTS: Of 426 patients randomly assigned, 48% were aged >70 years, mean LVEF was 25% (SD, 8) and median plasma N-terminal pro-brain natriuretic peptide was 3,070 pg/ml (interquartile range 1,285 to 6,749 pg/ml). During 240 days of follow-up, 19.5%, 15.9%, and 12.7% of days were lost as the result of death or hospitalization for UC, NTS, and HTM, respectively (no significant difference). The number of admissions and mortality were similar among patients randomly assigned to NTS or HTM, but the mean duration of admissions was reduced by 6 days (95% confidence interval 1 to 11) with HTM. Patients randomly assigned to receive UC had higher one-year mortality (45%) than patients assigned to receive NTS (27%) or HTM (29%) (p = 0.032). CONCLUSIONS: Further investigation and refinement of the application of HTM are warranted because it may be a valuable role for the management of selected patients with heart failure. PMID- 15893184 TI - Improving the outcomes of heart failure care: putting technology second. PMID- 15893185 TI - Relationship between B-type natriuretic peptides and pulmonary capillary wedge pressure in the intensive care unit. AB - OBJECTIVES: We examined whether B-type natriuretic peptides (BNP) can serve as noninvasive markers of pulmonary capillary wedge pressure (PCWP) in the setting of critical illness. BACKGROUND: The BNP and N-terminal pro-B-type natriuretic peptide (NT-proBNP) are highly correlated with left ventricular (LV) filling pressures in patients with depressed LV systolic function. However, their relationship to PCWP in a heterogeneous intensive care unit (ICU) population has not been established. METHODS: We prospectively studied 40 patients in the ICU requiring invasive hemodynamic monitoring. Hemodynamics were recorded simultaneously with blood sampling for BNP and NT-proBNP. RESULTS: The BNP (median 420 pg/ml) and NT-proBNP (median 3,304 pg/ml) levels were markedly elevated, but weakly correlated with PCWP (BNP, r = 0.40, NT-proBNP, r = 0.32) and other cardiac parameters. Peptide levels were approximately four-fold greater in patients with impaired (estimated glomerular filtration rate [eGFR] <60 ml/min) versus normal (eGFR >60 ml/min) renal function, despite similar PCWP, cardiac index, and LV ejection fraction. In addition, both BNP and NT-proBNP showed stronger correlations with PCWP in patients with preserved (BNP, r = 0.58, NT-proBNP, r = 0.73) versus impaired renal function (BNP, r = 0.48, NT-proBNP, r = 0.34). Interaction terms between eGFR and BNP (p = 0.06) and NT-proBNP (p = 0.04) suggest that eGFR modulates the relationship of these peptides to filling pressures. CONCLUSIONS: The BNPs are markedly elevated, yet show only weak correlations to PCWP in ICU patients requiring invasive hemodynamic monitoring. Thus, a single value for BNP or NT-proBNP may not be a clinically useful noninvasive marker of filling pressures in the critically ill patient. This appears to be especially true in patients with impaired renal function. PMID- 15893186 TI - Severe symptomatic tricuspid valve regurgitation due to permanent pacemaker or implantable cardioverter-defibrillator leads. AB - OBJECTIVES: We report a series of patients with severe tricuspid valve regurgitation due to a permanent pacemaker (PPM) or implantable cardioverter defibrillator (ICD) lead. BACKGROUND: Severe tricuspid regurgitation caused by a PPM or ICD lead is an under-recognized but treatable etiology of severe right heart failure. METHODS: We reviewed the records of 41 patients who underwent tricuspid valve operation for severe tricuspid regurgitation caused by previously placed PPM or ICD leads. RESULTS: During surgery, severe tricuspid regurgitation was found to be caused by the PPM or ICD leads in all 41 patients. There was a perforation of the tricuspid valve leaflet by the PPM or ICD lead in 7 patients, lead entanglement in the tricuspid valve occurred in 4 patients, lead impingement of the tricuspid valve leaflets occurred in 16 patients, and lead adherence to the tricuspid valve occurred in 14 patients. The septal leaflet was most often perforated (6 of 7). In the preoperative evaluation, valve malfunction due to the PPM or ICD lead was diagnosed preoperatively in only 5 of 41 (12%) patients by transthoracic echocardiography. All patients underwent successful tricuspid valve operation (22 tricuspid valve replacement), with one perioperative death occurring. During follow-up (range, 1 to 99 months), there was one patient who died from left-sided heart failure and three patients died of other causes. The remaining patients showed improvement in signs and symptoms of heart failure. CONCLUSIONS: Damage to the tricuspid valve by PPM or ICD leads may result in severe symptomatic tricuspid regurgitation and may not be overtly visualized by echocardiography. This etiology should be considered when evaluating patients with severe right heart failure after PPM or ICD implantation. PMID- 15893187 TI - Prognostic value of lung sestamibi uptake in myocardial perfusion imaging of patients with known or suspected coronary artery disease. AB - OBJECTIVES: We sought to determine whether lung uptake of technetium-99m (99mTc) based myocardial perfusion tracers predicts cardiac events. BACKGROUND: Increased lung uptake of thallium-201 during myocardial perfusion scintigraphy can predict important clinical outcomes. It is unclear whether lung uptake of 99mTc-based myocardial perfusion tracers can be used in a similar way. METHODS: Stress lung to-heart ratio (sLHR) was determined in 718 patients undergoing 99mTc-sestamibi single-photon emission computed tomographic stress imaging. The primary outcome was acute myocardial infarction or death. RESULTS: During a mean follow-up of 5.6 years, a primary end point occurred in 114 patients (16%). The sLHR was significantly greater in those with an adverse outcome (p < 0.00001). The likelihood of an adverse outcome increased by a factor of 1.5 (95% confidence interval 1.2 to 1.7) for each standard deviation increase in sLHR after adjustment for all other variables. The sLHR provided a small but significant improvement in risk stratification when added to clinical, stress test, perfusion, and left ventricular volume information (global chi-square 168.6 vs. 150.7, p < 0.00001). CONCLUSIONS: Stress LHR is an adjunctive prognostic measure in patients with known or suspected coronary artery disease. PMID- 15893188 TI - Evaluation of the accuracy of gadolinium-enhanced cardiovascular magnetic resonance in the diagnosis of cardiac sarcoidosis. AB - OBJECTIVES: This study analyzed the accuracy of gadolinium-enhanced cardiovascular magnetic resonance (CMR) for the diagnosis of cardiac sarcoidosis (CS). BACKGROUND: The diagnosis of CS was made according to the guidelines of the Japanese Ministry of Health and Welfare (1993); CMR has not been incorporated into the guidelines, and the diagnostic accuracy of CMR for the diagnosis of CS has not yet been evaluated. METHODS: We performed an analysis of 12-lead electrocardiograms (ECGs), 24-h ambulatory ECGs, echocardiograms, thallium scintigrams, and gadolinium-enhanced CMR studies in 58 biopsy-proven pulmonary sarcoidosis patients assessed for CS. The diagnostic accuracy of CMR for CS was determined using modified Japanese guidelines as the gold standard. RESULTS: The diagnosis of CS was made in 12 of 58 patients (21%); CMR revealed late gadolinium enhancement (LGE), mostly involving basal and lateral segments (73%), in 19 patients. In 8 of the 19 patients, scintigraphy was normal, while patchy LGE was present. The sensitivity and specificity of CMR were 100% (95% confidence interval, 78% to 100%) and 78% (95% confidence interval, 64% to 89%), and the positive and negative predictive values were 55% and 100%, respectively, with an overall accuracy of 83%. CONCLUSIONS: In patients with sarcoidosis, CMR is a useful diagnostic tool to determine cardiac involvement. New diagnostic guidelines should include CMR. PMID- 15893189 TI - Impact of pulmonary hypertension on the outcomes of noncardiac surgery: predictors of perioperative morbidity and mortality. AB - OBJECTIVES: We sought to determine the predictors of short-term morbidity and mortality (< 30 days) after noncardiac surgery in patients with pulmonary hypertension (PH). BACKGROUND: Pulmonary hypertension is considered to be a significant preoperative risk factor. METHODS: The PH and surgical data bases were matched from 1991 to 2003. Patients were excluded if PH was secondary to left heart disease, not present before surgery, or the procedure involved cardiopulmonary bypass. Univariate and multivariate logistic regression analyses were used to identify variables associated with short-term morbidity and mortality. RESULTS: Of 1,276 patients in the PH database, 145 patients (73% female) met all study criteria. The mean age (+/-SD) was 60.1 +/- 16.0 years. Right ventricular systolic pressure (RVSP) (mean +/- SD) on the two-dimensional echocardiogram was 68 +/- 21 mm Hg. There were 60 patients (42%) who experienced one or more short-term morbid event(s) (1.8 events/patient experiencing any event). A history of pulmonary embolism (p = 0.01), New York Heart Association functional class > or = II (p = 0.02), intermediate- to high-risk surgery (p = 0.04), and duration of anesthesia > 3 h (p = 0.04) were independent predictors of short-term morbidity. There were 10 early deaths (7%). A history of pulmonary embolism (p = 0.04), right-axis deviation (p = 0.02), right ventricular (RV) hypertrophy (p = 0.04), RV index of myocardial performance > or = 0.75 (p = 0.03), RVSP/systolic blood pressure > or = 0.66 (p = 0.01), intraoperative use of vasopressors (p < 0.01), and anesthesia when nitrous oxide was not used (p < 0.01) were each associated with postoperative mortality. CONCLUSIONS: In patients with PH undergoing noncardiac surgery with general anesthesia, specific clinical, diagnostic, and intraoperative factors may predict worse outcomes. PMID- 15893190 TI - The tyrosine phosphatase, SHP-1, is a negative regulator of endothelial superoxide formation. AB - OBJECTIVES: We investigated the role of SH2-domain containing phosphatase-1 (SHP 1) in endothelial reduced nicotinamide adenine dinucleotide (phosphate) (NAD[P]H) oxidase-dependent oxidant production. BACKGROUND: Superoxide (O2*-) generation by endothelial NAD(P)H-oxidase promotes endothelial dysfunction and atherosclerosis. Signaling pathways that regulate NAD(P)H-oxidase activity are, however, poorly understood. METHODS: SH2-domain containing phosphatase-1 was inhibited using site directed magnetofection of antisense oligodesoxynucleotides (AS-ODN) or short interfering ribonucleic acid (siRNA) in vitro in human umbilical vein endothelial cells (HUVEC) and in isolated hamster arteries; O2*- was measured by cytochrome c reduction in vitro. Activities of NAD(P)H-oxidase activity, phosphatidyl-inositol 3-kinase (PI3K), and SHP-1 were assessed by specific assays; Rac1 activation was assessed by a pull-down assay. RESULTS: Basal endothelial O2*- release was enhanced after inhibition of endothelial SHP-1 (p < 0.01), which could be prevented by specific inhibition of NAD(P)H-oxidase (p < 0.01); SHP-1 activity was high under basal conditions, further increased by vascular endothelial growth factor (10 ng/ml, p < 0.05), and abolished by SHP-1 AS-ODN treatment (p < 0.01), which also increased NAD(P)H-oxidase activity 3.3-fold (p < 0.01). Vascular endothelial growth factor also induced O2*- release (p < 0.01), which was even more enhanced when SHP-1 was knocked down (p < 0.05). The effect of SHP-1 was mediated by inhibition of PI3K/Rac1-dependent NAD(P)H-oxidase activation (p < 0.01); SHP-1 AS-ODN augmented tyrosine phosphorylation of the p85 regulatory subunit of PI3K (p < 0.05) and Rac1 activation. The latter was prevented by wortmannin, a blocker of PI3K. CONCLUSIONS: In HUVEC, SHP-1 counteracts basal and stimulated NAD(P)H-oxidase activity by negative regulation of PI3K-dependent Rac1 activation; SHP-1 thus seems to be an important part of endothelial antioxidative defense controlling the activity of the O2(*-)-producing NAD(P)H-oxidase. PMID- 15893191 TI - Cyclooxygenase-1 mediates the final stage of morphine-induced delayed cardioprotection in concert with cyclooxygenase-2. AB - OBJECTIVES: We sought to investigate the time course of morphine-induced delayed cardioprotection and examine the role of cyclooxygenase (COX) in this cardioprotective effect. BACKGROUND: Cyclooxygenase-2 has been shown to be essential for the delayed cardioprotection induced by ischemic preconditioning and delta-opioid agonists. METHODS: Male mice were subjected to 45 min of coronary artery occlusion followed by 120 min of reperfusion. Expressions of COX 2 and COX-1 were assessed by Western blotting, and the myocardial prostaglandin (PG)E2 and 6-keto-PGF(1-alpha) contents were measured using enzyme immunoassays. RESULTS: A powerful infarct-sparing effect appeared 24 and 48 h after morphine preconditioning and faded after 72 h. After 24 h, the anti-infarct effect was associated with enhanced myocardial levels of COX-2, PGE2, and 6-keto-PGF(1 alpha), and no changes in COX-1 protein levels were found. Cardioprotection and increases in PGE2 and 6-keto-PGF(1-alpha) were completely abolished by the COX-2 selective inhibitor NS-398 and the non-selective COX inhibitor indomethacin, whereas the COX-1-selective inhibitor SC-560 had no effect. After 48 h, up regulation of myocardial PGE2 and 6-keto-PGF(1-alpha) was also observed, and COX 1 expression was enhanced markedly, but only a slight increase in COX-2 expression was apparent. Cardioprotection and the increases in PGE2 and 6-keto PGF(1-alpha) 48 h after morphine administration were abrogated only by indomethacin, and not by SC-560 or NS-398. CONCLUSIONS: Morphine confers delayed cardioprotection via a COX-dependent pathway; COX-2 is essential for the cardioprotection observed in the initial stage (24 h), whereas, in the final stage (48 h), cardioprotection is mediated by COX-1 in concert with COX-2. PMID- 15893192 TI - Dietary linolenic acid and adjusted QT and JT intervals in the National Heart, Lung, and Blood Institute Family Heart study. AB - OBJECTIVES: The goal of this study was to examine whether higher consumption of total linolenic acid was associated with rate-adjusted QT and JT intervals (QTrr and JTrr, respectively). BACKGROUND: Higher intake of fish omega-3 fatty acids and plant omega-3 such as alpha-linolenic acid is associated with lower risk of myocardial infarction. While long-chain omega-3 can inhibit ventricular arrhythmia, it is not known whether alpha-linolenic acid influences ventricular repolarization. METHODS: We studied 3,642 subjects from the National Heart, Lung, and Blood Institute Family Heart study who were free of myocardial infarction, left ventricular hypertrophy, pacemaker, and with QRS <120 ms. We used the 95th percentile of the gender-specific distribution of QTrr and JTrr to define abnormally prolonged repolarization. Within each gender, we created age- and energy-adjusted tertiles of linolenic acid and used regression models for analyses. RESULTS: Mean age was 50 years, and average intake of total linolenic acid was 0.74 g/day. There was an inverse association between consumption of linolenic acid and QTrr and JTrr (p for trend 0.001 and 0.0005, respectively). From the lowest (reference) to the highest gender-, age-, and energy-adjusted tertile of linolenic acid, multivariable adjusted odds ratios for prolonged QTrr were 1.0, 0.74 (95% confidence interval [CI] 0.57 to 0.96), and 0.59 (95% CI 0.44 to 0.77), respectively (p for trend 0.0003). Corresponding values for JTrr were 1.0, 0.73 (95% CI 0.52 to 1.03), and 0.59 (95% CI 0.40 to 0.87), respectively (p for trend 0.009). Exclusion of subjects taking drugs known to influence QT did not influence this association. CONCLUSIONS: Higher intake of dietary linolenic acid might be associated with a reduced risk of abnormally prolonged repolarization in men and women. PMID- 15893193 TI - N-3 Fatty acids for the prevention of atrial fibrillation after coronary artery bypass surgery: a randomized, controlled trial. AB - OBJECTIVES: The aim of this study was to assess the efficacy of preoperative and postoperative treatment with n-3 polyunsaturated fatty acids (PUFAs) in preventing the occurrence of atrial fibrillation (AF) after coronary artery bypass graft surgery (CABG). BACKGROUND: Postoperative AF is a common complication of CABG. There is growing clinical evidence that PUFAs have cardiac antiarrhythmic effects. METHODS: A total of 160 patients were prospectively randomized to a control group (81 patients, 13 female, 64.9 +/- 9.1 years) or PUFAs 2 g/day (79 patients, 11 female, 66.2 +/- 8.0 years) for at least 5 days before elective CABG and until the day of discharge from the hospital. The primary end point was the development of AF in the postoperative period. The secondary end point was the hospital length of stay after surgery. All end points were independently adjudicated by two cardiologists blinded to treatment assignment. RESULTS: The clinical and surgical characteristics of the patients in the two groups were similar. Postoperative AF developed in 27 patients of the control group (33.3%) and in 12 patients of the PUFA group (15.2%) (p = 0.013). There was no significant difference in the incidence of nonfatal postoperative complications, and postoperative mortality was similar in the PUFA-treated patients (1.3%) versus controls (2.5%). After CABG, the PUFA patients were hospitalized for significantly fewer days than controls (7.3 +/- 2.1 days vs. 8.2 +/- 2.6 days, p = 0.017). CONCLUSIONS: This study first demonstrates that PUFA administration during hospitalization in patients undergoing CABG substantially reduced the incidence of postoperative AF (54.4%) and was associated with a shorter hospital stay. PMID- 15893194 TI - Immigration policy and health care. PMID- 15893196 TI - Detection of pathologic or physiologic left ventricular remodeling in athletes. PMID- 15893197 TI - World Trade Center attack and cardiac events: fact or fear? PMID- 15893199 TI - The optimal low-density lipoprotein is 50 to 70 mg/dl. PMID- 15893200 TI - Improved outcomes of pediatric dilated cardiomyopathy and heart transplantation. PMID- 15893202 TI - Cell transplantation and fibrin matrix. PMID- 15893205 TI - Implementing shared decision making in diverse health care systems: the role of patient decision aids. PMID- 15893206 TI - Information needs and sources of information among cancer patients: a systematic review of research (1980-2003). AB - Understanding what cancer patients need to know and from whom they receive information during the course of care is essential to ensuring quality care. We reviewed 112 articles published from 1980 to 2003 and developed a typology summarizing cancer patients' information needs and the sources from which they receive information. The majority of articles focused on information needs and sources during the diagnosis and treatment phase. Thus, the most frequent information need was treatment-related (38.1%). The most frequent information source was health professionals (27.3%). We examined patients' information needs and sources along the continuum of care and found that during diagnosis and treatment, information needs about the stage of disease, treatment options, and side-effects of treatment were prominent; during post-treatment, patients continued to need information about treatment, and information about recovery was also important. Future research should examine cancer patients' information needs and sources throughout their cancer journey. PMID- 15893207 TI - Further analysis of a doctor-patient nonverbal communication instrument. AB - This study examines the reliability and validity of the relational communication scale for observational measurement (RCS-O) using a random sample of 80 videotaped interactions of medical students interviewing standardized patients (SPs). The RCS-O is a 34-item instrument designed to measure the nonverbal communication of physicians interacting with patients. The instrument was applied and examined in two different interview scenarios. In the first scenario (year 1), the medical student's interview objective is to demonstrate patient-centered interviewing skills as the SP presents with a psychosocial concern. In the second scenario (year 3), the student's interview objective is to demonstrate both doctor-centered and patient-centered skills as the SP presents with a case common in primary care. In the year 1 scenario, 19 of the 34 RCS-O items met acceptable levels of inter-rater agreement and reliability. In the year 3 scenario, 26 items met acceptable levels of inter-rater agreement and reliability. Factor analysis indicated that in both scenarios each of the four primary relational communication dimensions was salient: intimacy, composure, formality, and dominance. Measures of correlation and differences involving the RCS-O dimensions and structural features of the interviews (e.g., number of questions asked by the medical student) are examined. PMID- 15893208 TI - Implementation of a professional program in Finnish community pharmacies in 2000 2002. AB - The aim was to assess implementation of a national 4-year project to promote professional strategy in Finnish community pharmacies during the 2.5 years of program's action in 2000-2002. A survey of random sample of 734 community pharmacists in Finland was conducted in 2002 (response rate 51%, n = 376). The questionnaire included an implementation scale and two open-ended questions on patient counselling. Many of the actions were not optimally implemented 2.5 years after starting the project. Some individual actions may have been taken, but the systematic long-term development process has not yet been started in many pharmacies. Future challenges in implementation will be a better commitment of the pharmacy owners to the goals of the national professional strategy and a better access to Internet-based drug information sources while counselling patients. PMID- 15893209 TI - Women's misconceptions about cancer screening: implications for informed decision making. AB - Informed decision-making about cancer screening requires that patients have a correct understanding of a test's purpose, benefits, and risks. Misconceptions, however, may be common. Semi-structured interviews were carried out and thematically coded using a purposive sample of 24 socioeconomically diverse white, African American, Latino and Chinese American women recruited from general medicine practices and community settings. Interviews focused on participants ideas related to cancer prevention and screening. Women expressed cancer-related beliefs characterized by inaccuracies, distortions, and over-simplifications. Many of these beliefs may go unrecognized in clinical settings yet have a profound influence on risk communication and, therefore, informed decision making. Effective communication depends, first, on clinicians and patients sharing an accurate understanding of background concepts such as "prevention," "screening," and "cancer." PMID- 15893210 TI - The decision evaluation scales. AB - There are several instruments to assess how patients evaluate their medical treatment choice. These are used to evaluate decision aids. Our objective is to investigate which psychological factors play a role when patients evaluate their medical treatment choices. A pool of 36 items was constructed, covering concepts such as uncertainty about and satisfaction with the decision, informed choice, effective decision making, responsibility for the decision, perceived riskiness of the choice, and social support regarding the decision. This pool was presented to patients at high risk for breast and ovarian cancer, awaiting a genetic test result, and facing the choice between prophylactic surgery or screening. Additional measures were assessed for validation purposes. Factor and Rasch analyses were used for factor and item selection. Construct validity of emerging scales was assessed by relating them with the additional measures. Three factors summarised the psychological factors concerning decision evaluation: Satisfaction Uncertainty, Informed Choice, and Decision Control. Reliabilities (Cronbach's alpha) of the three scales were 0.79, 0.85, and 0.75, respectively. Construct validity hypotheses were confirmed. The first two scales were similar to previously developed scales. Of these three scales, the Decision Control scale correlated most strongly with the well-being measures, was associated with partner's agreement and physician's preferences as perceived by patients, and with a negative emotional reaction to the information material. In conclusion, the Decision Control scale is a new scale to evaluate decision aids, and it appears to be rooted in health psychological theories. PMID- 15893211 TI - How making a risk estimate can change the feel of that risk: shifting attitudes toward breast cancer risk in a general public survey. AB - Counseling women about breast cancer risks has been found to decrease screening compliance. We investigated whether women's reactions to risk information are an artifact of requiring women to estimate the risk of breast cancer prior to receiving risk information. Three hundred and fifty-six women were randomized to either make or not make a risk estimate prior to receiving risk information. Outcome measures were participants' estimates of the average woman's breast cancer risk and their emotional response to the risk information. Women overestimated the lifetime risk of breast cancer (M = 46%). Women who made risk estimates felt more relieved about the risk and perceived the risk as being lower than women who did not make estimates (p's < 0.001). Asking people to estimate risks influenced their subsequent perceptions of the risk of breast cancer. PMID- 15893212 TI - Patients' perceived barriers to active self-management of chronic conditions. AB - Few studies have elicited barriers to patient self-management of chronic conditions, and only one concerned people with two or more conditions. To inform development of Homing in on Health (HioH), a home delivery variant of the Chronic Disease Self-Management Program (CDSMP), we conducted 10 focus groups involving 54 chronically ill people, 46 (85%) of whom had multiple conditions. The goals were to elicit perceived barriers to active self-management and to accessing self management support resources. Depression, weight problems, difficulty exercising, fatigue, poor physician communication, low family support, pain, and financial problems were the most frequently noted barriers to active self-management. The most common barriers to accessing self-management support resources were lack of awareness, physical symptoms, transportation problems, and cost/lack of insurance coverage. Our findings provided initial support for the Homing in on Health approach, since many of the barriers identified may be more amenable to home based intervention than to centralized, facility-based programs. PMID- 15893213 TI - Preferences for medical collaboration: patient-physician congruence and patient outcomes. AB - Patient participation in medical care and in decision-making is generally viewed as a precursor to positive health outcomes. Patient participation is not always possible or desirable, however, and not all patients want to take an active part in their own medical care. This study examines the degree to which physician patient congruence in preference for patient involvement is related to self reported satisfaction, adherence, and health. Results indicate that when patients and their doctors share similar beliefs about patient participation, patient outcomes tend to be more positive, with highest satisfaction found in cases in which both patient and physician desire more patient involvement. PMID- 15893214 TI - It's good to talk: comparison of a telephone helpline and website for cancer information. AB - The Internet plays an important role in the information gathering process for people affected by cancer. This paper presents the results of a study comparing two sources of cancer information; a website and a telephone helpline. Data were captured on the use of one section of the UK charity CancerBACUP's website, and systematically compared to data collected from every fifth user of the charity's helpline. Subjects of enquiry for 994 telephone enquiries and 3096 web enquiries were comparable. The majority of enquiries to both services were about women and/or patients aged 50 or older. Website users were more likely than helpline users to request factual information and less likely to request information on sensitive topics. This study provides valuable information about the types of health information people seek from different sources and how the sources may complement each other. PMID- 15893215 TI - Doctors' use of euphemisms and their impact on patients' beliefs about health: an experimental study of heart failure. AB - Doctors often use a range of euphemisms as a means to facilitate communication in the consultation. The present experimental study aimed to assess whether GPs use or avoid the term 'heart failure' and to evaluate the relative impact of the term 'heart failure' versus their preferred euphemism on patients' beliefs about the illness. This two part study involved a cross sectional survey of GPs and an experimental study of patients' beliefs and was based on one General Practice in a semi-rural area of the UK. For the first part, 42 GPs completed a questionnaire about their preferred terms to describe symptoms of heart failure. The results showed that GPs rated the majority of euphemisms as preferable to the term 'heart failure'. Their preferred euphemism was 'fluid on your lungs as your heart is not pumping hard enough'. For the second part, 447 patients completed ratings of their beliefs about a condition, which was described as either 'heart failure' or the GPs' preferred euphemism. Patients who received the condition described as 'heart failure' believed that the illness would have more serious consequences for their life, that the problem would be more variable over time and that it would last for longer and reported feeling more anxious and depressed than those who received the condition described using the euphemism. GPs are encouraged to be open with their patients and to respect their experience. The choice of language, therefore, presents a dilemma for doctors. The term 'heart failure' may be in line with the current climate of openness but may evoke a more negative response from the patient. In contrast, a euphemism may be less open but more protective of the patient's experience. This study suggests that the area of heart failure may be one where GPs may chose to compromise openness for the sake of the patient's experience and that this fear of upsetting the patient is well founded. PMID- 15893216 TI - Physician use of a participatory decision-making style with children with ADHD and their parents. AB - The purpose of this study was to examine the extent to which physicians reported using a participatory decision-making (PDM) style with children and their parents during attention-deficit/hyperactivity disorder visits and the physician characteristics that were related to physician use of a PDM style. The survey was sent to a stratified random sample of 250 pediatricians and 250 family practitioners that were licensed and actively practicing in North Carolina. A second mailing of the survey was sent to non-responders approximately 3 weeks after the first mailing was sent. Approximately 47% of the physicians responded. Physicians were significantly more likely to rate themselves as more participatory with parents than with children. Younger physicians were more likely to use a participatory style with attention-deficit/hyperactivity disorder (ADHD) children. Physicians who rated themselves as using a more participatory style with parents, also rated themselves as being more participatory with children. Pediatricians and younger physicians were more likely to use a participatory style with parents of ADHD children. The majority of physicians believed that ADHD children could begin to contribute to decisions about the diagnosis and treatment of ADHD during medical visits from age 7 to 11 years. PMID- 15893217 TI - Development and validation of the Communication and Attitudinal Self-Efficacy scale for cancer (CASE-cancer). AB - We sought to develop a reliable and valid measure of patient self-efficacy within the context of productive communication and positive attitude for cancer patients. A set of 19 potential items for the Communication and Attitudinal Self Efficacy scale for cancer (CASE-cancer) was pilot tested with 50 cancer patients. Based on the pilot test, item valence was made consistent (i.e., all items worded positively) and the response scale was simplified. The CASE-cancer was then administered to 127 persons receiving cancer treatment at general oncology clinics in Shreveport, Louisiana and Chicago, Illinois. Psychometric analyses revealed three 4-item factors: understanding and participating in care, maintaining a positive attitude, and seeking and obtaining information. The CASE cancer proved to have high internal consistency and construct validity. Moreover, scale items performed similarly across literacy levels. The CASE-cancer is a psychometrically-sound tool that may provide new information on important mediating factors of cancer care. Our two-step approach to presenting response options may also provide a model for helping lower literate patients more accurately respond to survey items. PMID- 15893218 TI - Patient-centered care and breast cancer survivors' satisfaction with information. AB - Satisfaction with information is an important patient outcome and may be related to the physician's ability to elicit the patients' concerns, to consider the patients' psychosocial needs, and to involve patients in treatment decision making; these communication techniques have been described under the umbrella of "patient-centered." The aim of this study was to examine the relationship between patient-centered care and satisfaction with information among women with a history of breast cancer. We administered a questionnaire to 182 women who had completed treatment for breast cancer. Our findings suggest that, while breast cancer survivors are highly satisfied with information related to treatment, they are less satisfied with information related to the long-term physical, psychological, and social sequelae of the disease and its treatments. In multivariate analysis, patients' perception of patient-centered behaviors was strongly associated with patients' satisfaction with information. These results provide support for the theory that patient satisfaction is improved when physicians incorporate patient-centered behaviors into their care. PMID- 15893219 TI - Local anesthetics facilitate ion transport across lipid planar bilayer membranes under an electric field: dependence on type of lipid bilayer. AB - In order to elucidate the role of structural change of lipid membrane bilayer in the mode of action of local anesthetic, we studied the effects of local anesthetics, charged tetracaine and uncharged benzocaine, on ion permeability across various lipid planar bilayers (PC, mixed PC/PS (4/1, mol/mol); mixed PC/PE (1/1, mol/mol); mixed PC/SM (4/1, mol/mol)) under a constant applied voltage. The membrane conductances increased in the order of PC<40%) dose reduction was achieved in the lung in the prone treatment position. Similar dose reductions were also achieved for prone pectoral muscle, chest wall, and breast skin, irradiating less volume with high doses. Pectoral muscle, lumpectomy, and the breast skin showed prominent differences in shape and displacement in the prone compared with the supine position. CONCLUSION: CT-based treatment planning allowed calculations of volumetric dose distribution to the target and all sensitive structures with proper visualization and volumetric delineation of organs of interest. The treatment plan shows significant dose reduction may be possible in various sensitive structures if the patient is treated with HDR brachytherapy in the prone position. PMID- 15893268 TI - Feasibility and preliminary outcome of salvage combined HDR brachytherapy and external beam radiotherapy (EBRT) for local recurrences after radical prostatectomy. AB - PURPOSE: Feasibility of combined fractionated intensity modulated brachytherapy (IMBT) and external beam radiotherapy (EBRT) as well as the effect of local dose escalation was investigated in a non-randomized retrospective observation trial for histologically-proven macroscopic local recurrences of prostate cancer after radical prostatectomy. METHODS AND MATERIALS: Thirty-five patients with transrectal ultrasound (TRUS) detectable tumors were treated. Applied dose per IMBT fraction was 15 Gy, prescribed on the target (TRUS visible tumor) surface. For the first 21 patients, two fractions of IMBT were delivered in 2 weeks interval, complementary to 30 Gy EBRT to the small pelvis. Further, as second step of dose escalation, 14 patients were treated with 2 x 15 Gy IMBT combined with 40 Gy EBRT. The total treatment time was 4 and 5 weeks, respectively. RESULTS: PSA was decreased in 34 out of 35 patients post-therapeutically. After a mean follow-up of 27 months, 32 out of 35 patients are alive. However, in 67% of the patients, we observed postimplant PSA elevation with or without detectable local and/or systemic progress. The mean duration of biochemical non-evidence of disease (bNED) after radiation was 12 months for all patients (31% in the 30 Gy group and 42% in the 40 Gy group). No RTOG/EORTC grade III or IV side effects were registered during/after radiotherapy. CONCLUSION: Combined EBRT and IMBT boost of TRUS detectable recurrences of prostate cancer after radical prostatectomy seems to be a feasible method of salvage treatment. These early results need to be confirmed by further prospective randomized trials and by longer follow-up in all dose groups. PMID- 15893269 TI - Impact of selection of post-implant technique on dosimetry parameters for permanent prostate implants. AB - PURPOSE: To investigate the variability of prostate implant quality indices between three different methods of calculating the post-implant dose distribution. METHODS AND MATERIALS: In a study of 9 permanent prostate implant patients, post-implant dosimetry was carried out using three methods of identifying seed positions within the prostate volume: (1) prostate volumes defined by transrectal ultrasound (TRUS) immediately following implant were registered with shift-film defined seed positions, (2) seeds were identified directly from the post-implant TRUS images, and (3) CT was used to define seed positions and prostate volumes from images acquired at 41-65 days post-implant. For each method, the volume of prostate receiving 90%, 100%, and 150% of the prescribed dose (V90, V100, V150) and the dose delivered to 90% of the prostate volume (D90) were calculated. RESULTS: Post-implant TRUS volumes were within 15% of the preimplant TRUS volumes in 8 of the 9 patients investigated. The post implant CT volume was within 15% of the preimplant (TRUS) volume in only 3 of the 9 cases. The value of the dosimetry parameters was dependent on the method used and varied by 5-25% for V90, 5-30% for V100, 42-134% for V150, and 9-60% for D90. No simple relationship was found between change in volume and the resultant change in dosimetry parameter. Differences in dosimetry parameters due to source localization uncertainties was found to be small (< or = 10% for V100) when comparing methods (1) and (2). CONCLUSIONS: There are many uncertainties in the calculation of parameters that are commonly used to describe the quality of a permanent prostate implant. Differences in the parameters calculated were most likely a result of a combination of factors including uncertainties in delineating the prostate with different imaging modalities, differences in source identification techniques, and intraobserver variability. PMID- 15893270 TI - Feasibility report for retreatment of locally recurrent head-and-neck cancers by combined brachy-chemotherapy using frameless image-guided 3D interstitial brachytherapy. AB - PURPOSE: Brachytherapy re-irradiation may offer an alternative re-treatment of recurrent head-and-neck cancer even after previous full dose radiation therapy. The purposes of this study were to determine the feasibility and accuracy of frameless image-guided interstitial needle implantation. METHODS AND MATERIALS: Between January 2000 and March 2003, 14 patients with biopsy-proven locally recurrent head-and-neck-cancer were retreated after previous full dose irradiation with combined external beam-brachytherapy with concomitant chemotherapy. Brachytherapy needle implantation was virtually planned taking into account the surrounding risk structures. Needles were implanted using an adapted frameless navigation system. Chemoradiotherapy was followed by 2-4 courses of chemotherapy every fourth week starting 4 weeks after the end of brachytherapy. RESULTS: The 1- and 2-year local control rates were 78% and 57%, respectively. Local control was obtained in 8/14 patients. The actuarial 1- and 2-year survival rates were 83% and 64%, respectively. The median survival was 28 months after a median follow-up of 21 months (range, 8-53). Six weeks after brachytherapy, 1 patient developed localized soft tissue necrosis which did not require surgical intervention. No additional grade III or IV late toxicity was seen after re irradiation. Mean deviation of image-guided needle implantation was 3.4 mm for each needle (SD, 1.9 mm; range, 0.5-14 mm). The mean deviation of all needles of an implant was 4.3 mm (range, 2.3-8.6 mm). CONCLUSIONS: These data demonstrate that pulsed-dose-rate brachytherapy in combination with sequential chemotherapy is effective and safe in re-irradiation of locally recurrent oropharyngeal carcinomas and can be offered to patients with curative intent. Image guidance allows virtual planning and navigated implantation of brachytherapy needles with regard to optimized needle distribution and risk structures. PMID- 15893271 TI - Salvage of suboptimal prostate seed implantation: Reimplantation of underdosed region of prostate base. AB - PURPOSE: To demonstrate how a suboptimal (125)I prostate implant can be salvaged by reimplantation. METHODS AND MATERIALS: A (125)I implant was preplanned to deliver 150 Gy to the prostate of a patient with early stage prostate cancer. A CT scan at 35 days postimplant indicated that V(100) and D(90) were 46% and 49 Gy, respectively. The cause was a systematic source placement error that left the base significantly underdosed. A reimplantation of the underdosed region was planned by superimposing a template grid onto the 35-day postimplant CT scan images and digitizing them into the treatment planning computer as if they were TRUS images. The reimplantation was carried out under fluoroscopy guidance so that the initial implant was visible. RESULTS: The reimplantation increased V(100) and D(90) to 98% and 201 Gy, respectively. The misplaced seeds resulted in a high dose to the apical region and urethra, which was further increased by the reimplantation. The patient experienced increased urinary morbidity, which was relieved by medication. CONCLUSION: It is feasible to salvage a suboptimal prostate seed implant by reimplanting the underdosed regions under fluoroscopy guidance based on a plan generated from the postimplant CT scan. PMID- 15893272 TI - (32)P radioisotope therapy for recurrent pilocytic astrocytoma. AB - (32)P is a pure beta-emitter that has a depth of penetration of 2-3 mm and can be useful in the treatment of cystic lesions. Its effectiveness in the treatment of a selected brain tumor is illustrated here. PMID- 15893273 TI - The optical structures of animal eyes. PMID- 15893274 TI - Colonic targeting of aminosalicylates for the treatment of ulcerative colitis. AB - Aminosalicylates (5-aminosalicylic acid) represent drugs of first choice in the treatment of ulcerative colitis. Two different therapeutic approaches have been employed to target the active 5-aminosalicylic acid to its site of action. Either inactive azo-prodrugs (e.g. sulfasalazine, olsalazine, balsalazide) or special galenic formulations have been developed for topical delivery of 5-aminosalicylic acid to the colon. However, as intestinal physiology, the extent of ulcerative colitis as well as drug disposition demonstrate large interindividual differences, acute healing rates (40-80%) and the maintenance of remission are quite variable. Apparently, therapeutic effects depend on local concentrations of 5-aminosalicylic acid in the colonic mucosa whereas systemic drug exposure might be one determinant of side effects. In general, 5-aminosalicylic acid is well tolerated and withdrawal from therapy is rare. Following administration of azo prodrugs (e.g. olsalazine), lower plasma concentrations and higher delivery into the colon of 5-aminosalicylic acid can be observed in comparison to special galenic formulations of 5-aminosalicylic acid. Whether such changes in drug disposition will affect therapeutic efficacy remains to be proved by clinical data. Consequently, selection of a particular agent should be based primarily on clinical efficacy, profile of adverse effects, patients' acceptance and economic considerations. PMID- 15893275 TI - The role of CT in coeliac disease. Methodology of the studies assessing diagnostic accuracy. PMID- 15893276 TI - The use of immunosuppressive drugs in IBD: RAND or random choice? PMID- 15893277 TI - Implications of corpus gastritis, atrophy and cyclooxygenase in the development of gastric erosions after curing Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori eradication decreases recurrence of peptic ulcers with marked improvement in histological inflammation, but gastric mucosal injuries may be developed even after eradication. PURPOSE: To investigate the mechanisms responsible for the development of gastric erosions after eradication, we analysed the relationship between clinicopathological risk factors and the occurrence of gastric erosion after curing H. pylori infection. PATIENTS: Sixty patients underwent endoscopy before, and 3, 6 and 12 months after the completion of H. pylori eradication. METHODS: Risk factors associated with the development of gastric erosions after eradication were assessed by multivariate analysis, and cyclooxygenase-1 and -2 immunoreactivity was histologically examined in the gastric mucosa before and after eradication. RESULTS: The cumulative prevalence of gastric erosions after H. pylori eradication was 38.3% within 1 year. Using multivariate analysis, corpus gastritis scores (inflammation score+activity score), corpus atrophy scores and an age of more than 50 years were found to be independent factors associated with the development of gastric erosion after eradication with odds ratios of 7.39, 0.13 and 5.00, respectively. Cyclooxygenase 2 immunoreactivity of the corpus was decreased for the non-erosion group after eradication, but not for the erosion group. CONCLUSIONS: Severe gastritis or less severe atrophy in oxyntic glands but not in pyloric glands before eradication may be involved in the development of gastric erosions after curing H. pylori infection. PMID- 15893278 TI - Abdominal CT findings may suggest coeliac disease. AB - BACKGROUND: Coeliac disease is the most common gastrointestinal immunological disorder in the western countries. Many adult patients present non-specific symptoms and signs of malabsorption such as chronic diarrhoea, anaemia, weight loss and abdominal distention. In non-specific and doubtful conditions, computed tomography is often the first medical examination performed. In a clinical practice, a critical review of computed tomography signs is therefore mandatory. AIMS: To evaluate the abdominal computed tomography findings, which are useful to suggest the presence of coeliac disease in adult patients. PATIENTS AND METHODS: The computed tomography studies of 28 coeliac patients were reviewed, looking for any intestinal and extraintestinal abnormality. The computed tomography findings evaluated were: abnormalities of intestinal fold pattern, bowel dilatation, fluid and air excess, duodenal abnormalities, intestinal intussusception, bowel wall thickening, lymphadenopathy, ascites, intestinal stenosis, mesenteric vascular changes. The abdominal computed tomography of a group of 30 normal subjects was also analysed. RESULTS: Intestinal fold pattern abnormalities were seen in 23/28 patients. Intestinal dilatation was seen in 21/28. Fluid excess in 18/28 and lymphadenopathy was seen in 12/28 patients; engorgement of mesenteric vessels in 7/28. Bowel wall thickening was observed in 6/28 patients and transient intussusception was observed in 6/28 patients. Increased air content within the bowel in 4/28 and ascites in 2/28 patients. Bowel dilatation together with fluid excess was observed in 18/28 patients. None of the above mentioned abnormalities abnormalities were seen in normal subjects. CONCLUSIONS: Data of the present study show that several abdominal computed tomography findings may be seen in coeliac adult patients; these findings should be taken into consideration with a high in level of suspicion by radiologists, to avoid diagnostic delay and unnecessary diagnostic and therapeutic procedures. PMID- 15893279 TI - Appropriateness of immunosuppressive drugs in inflammatory bowel diseases assessed by RAND method: Italian Group for IBD (IG-IBD) position statement. AB - INTRODUCTION: Despite the explosion of biological therapies, the old immunosuppressants continue to play a pivotal role in the management of inflammatory bowel diseases. AIM: To assess the appropriateness of immunosuppressants-azathioprine, 6-mercaptopurine, methotrexate, cyclosporine A, tacrolimus (FK506), mycophenolate mofetil and thalidomide-in the treatment of inflammatory bowel disease by using RAND/University of California Appropriateness Method. METHODS: The RAND method consists of a combination of evidence from the literature and experts' opinions. Appropriateness has been defined to mean that the expected health benefit exceeds the expected negative consequences by a sufficiently wide margin. A panel of 10 experts from the Italian Group for Inflammatory Bowel Disease has rated, in two rounds, on a scale from 1 to 9, the appropriateness of each indication selected by the Promoter Centre, on the basis of their own clinical experience. An indication was considered appropriate if the median of the panelists' ratings fell within the area 7-9, inappropriate in the area 1-3 and uncertain in the area 4-6. A total of 2781 indications were grouped into 13 categories (mild to moderate Crohn's disease; severe Crohn's disease; fistulizing Crohn's disease; steroid-dependant and -resistant Crohn's disease; maintenance of remission induced by medical treatment in Crohn's disease; maintenance of remission induced by surgery in Crohn's disease; mild to moderate ulcerative colitis; severe ulcerative colitis; steroid-dependant and -resistant ulcerative colitis; maintenance of remission induced by medical treatment in ulcerative colitis; extra-intestinal manifestations in inflammatory bowel disease; pregnancy and inflammatory bowel disease; azathioprine-resistant or intolerant inflammatory bowel disease patients). RESULTS: Of the 2781 scenarios, 212 (7.6%) were rated appropriate, 645 (23.2%) uncertain and 1924 (69.2%) inappropriate. The most relevant results were: in steroid-dependant or -resistant Crohn's disease, azathioprine, 6-mercaptopurine and methotrexate were defined as appropriate in 25 (86.2%) and 14 (48.3%) of the 29 scenarios respectively; in Crohn's disease, azathioprine and 6-mercaptopurine were defined as appropriate combined with Infliximab (bridge therapy); in steroid-dependant or -resistant ulcerative colitis, azathioprine and 6-mercaptopurine were defined as appropriate in 45 (77.6%) out of 58 scenarios, while methotrexate was defined appropriate only after previous azathioprine failure; in severe ulcerative colitis, cyclosporine A was defined as appropriate only after previous failure with steroids; in azathioprine-intolerant or -resistant inflammatory bowel disease patients, methotrexate was appropriate in 20 (66.7%) out of 30 scenarios; it is inappropriate to stop azathioprine treatment before conception in the presence of active disease. The use of FK506, mycophenolate mofetil and Thalidomide resulted as inappropriate or uncertain. CONCLUSIONS: Results of this study show that only azathioprine, 6-mercaptopurine and methotrexate are appropriate in the treatment of inflammatory bowel diseases. Cyclosporine A was found to be appropriate only in severe ulcerative colitis after the failure of steroids. FK506, mycophenolate mofetil and Thalidomide resulted as inappropriate but experience with these agents is somewhat limited. PMID- 15893280 TI - Prevalence, detection rate and outcome of cytomegalovirus infection in ulcerative colitis patients requiring colonic resection. AB - OBJECTIVE: To determine the prevalence of cytomegalovirus infection in patients with steroid-refractory ulcerative colitis who required colonic resection, and to assess its possible association with the use of immunosuppressive and steroid treatment and outcome after colectomy. PATIENTS AND METHODS: The study included surgical specimens and related pre-operative endoscopic biopsy specimens of 77 consecutive ulcerative colitis patients (34 females) who underwent colectomy because of intractable steroid-refractory ulcerative colitis (55 patients), toxic megacolon (6 patients), dysplasia or cancer (7 patients) or loss of function of the colon (9 patients). Clinical features and current and past treatments were analysed. Haematoxylin and eosin and specific immunohistochemical staining for cytomegalovirus were used to detect inclusion bodies in all specimens. RESULTS: Cytomegalovirus infection was found in 15 of 55 steroid-refractory ulcerative colitis patients (27.3%) and in 2 of 22 non-refractory patients (9.1%) (p=0.123). Only six patients had positive staining for cytomegalovirus in pre-operative endoscopic biopsy specimens. Detection of cytomegalovirus inclusion in biopsy specimens was not related to the number of biopsies or to time that had elapsed since colonoscopy and index surgery. Cytomegalovirus-positive patients were more likely to be on systemic corticosteroids (p=0.03). In contrast, current use and duration of immunosuppressive treatment, number of steroid cycles since diagnosis and in the last year, as well as chronic use of steroid in the last year were not significantly related to cytomegalovirus infection. Cytomegalovirus-positive patients did not receive antiviral therapy following proctocolectomy but did not show endoscopic or histological cytomegalovirus reactivation in the ileo-anal pouch and in the remaining bowel. CONCLUSIONS: Cytomegalovirus infection is frequently found in surgical specimens of patients with steroid-refractory ulcerative colitis and is more likely in patients on corticosteroid treatment. Cytomegalovirus infection is frequently unrecognised in pre-operative biopsy specimens, thus raising concerns about the accuracy of the available diagnostic tools. Unrecognised and untreated cytomegalovirus infection does not affect the outcome of ulcerative colitis patients following proctocolectomy. PMID- 15893281 TI - Immune activation and nutritional status in adult Crohn's disease patients. AB - BACKGROUND: Recent attention focused on the effect of inflammatory cytokines on intermediary metabolism contributing to the nutritional disturbances observed in acute or chronic inflammatory diseases. AIMS: To examine the interactions between immune activation and nutritional parameters in adult Crohn's disease patients. PATIENTS AND METHODS: We analysed anthropometric and biochemical nutritional parameters in 40 Crohn's disease patients and 26 healthy controls, and related them to inflammatory and immune markers. RESULTS: Weight, body mass index, mid arm circumference, triceps skinfold thickness, as well as albumin, transthyretin, retinol binding protein, insulin growth factor-I and Vitamin A were significantly decreased in Crohn's disease patients and negatively correlated to disease activity. By contrast, erythrocyte sedimentation rate, fibrinogen, C-reactive protein, alpha1-acylglycoprotein, soluble receptor of interleukin-2, blood neopterin, tumour necrosis factor-alpha and interleukin-1beta concentrations were significantly higher in patients and positively correlated to disease activity. Nutritional parameters and acute phase reactants were linked to tumour necrosis factor-alpha and interleukin-1beta concentrations, and markers of nutritional status were negatively correlated to positive acute phase reactants. CONCLUSIONS: In Crohn's disease, inflammatory cytokines appear partly responsible for decreased nutritional status. Thus, nutritional intervention to correct nutritional (in particular protein) depletion, and/or therapeutic intervention reducing inflammation and therefore restoring adequate nutritional proteins synthesis, appears a major therapeutic goal in active Crohn's disease. PMID- 15893282 TI - Gastrointestinal symptoms in infancy: a population-based prospective study. AB - BACKGROUND: During the first months of life, infants can suffer from many 'minor' gastroenterological disturbances. However, little is known about the frequency of these problems and the factors which predispose or facilitate their onset. AIMS: (a) To ascertain the frequency of the most common gastrointestinal symptoms in infants during the first 6 months after birth; (b) to evaluate the influence of some variables on the onset of the symptoms. STUDY DESIGN AND PATIENTS: Each of the 150 paediatricians distributed throughout Italy followed 20 consecutive infants from birth to 6 months. 2879 infants (1422 f, 1457 m) concluded the study. The presence of the following symptoms was evaluated: constipation, diarrhoea, vomiting, regurgitation, failure to thrive and prolonged crying fits (colic). Symptoms were recorded whenever the parents requested a clinical check up or during a set monthly examination. RESULTS: 1582/2879 (54.9%) infants suffered from one of the gastrointestinal symptoms. Regurgitation was the most common disturbance (present in 23.1% of infants), followed by colic (20.5%), constipation (17.6%), failure to thrive (15.2%), vomiting (6%) and diarrhoea (4.1%). Low birth weight was the factor most frequently associated with the onset of gastrointestinal symptoms, followed by low gestational age. Feeding habits did not influence the onset of symptoms, with the exception of constipation, which was linked to a low frequency of breast-feeding. Ninety-three infants (3.2%) were hospitalised for one or more of the gastrointestinal symptoms which were considered. During the whole study period the type of formula-milk was changed in 60% of the infants with one or more gastrointestinal symptoms, and in 15.5% of the infants who did not suffer from any gastrointestinal troubles. CONCLUSIONS: Gastrointestinal symptoms are very common in infants during the first 6 months after birth. These symptoms required hospitalisation only in a small percentage of cases, but led to the prescription of a 'dietary' milk formula in approximately 60% of the cases. Low birth weight and low gestational age were the main factors influencing the onset of the symptoms. PMID- 15893283 TI - Telomerase activity in proximal and distal gastric neoplastic and preneoplastic lesions using immunohistochemical detection of hTERT. AB - BACKGROUND: The incidence of distal (corpus and antrum) gastric adenocarcinoma is decreasing with a simultaneous increase in incidence of proximal (cardia) adenocarcinoma. Epidemiological studies suggest that they may represent different diseases but corroborative molecular data are scarce. Intestinal metaplasia may have a lower malignant potential in the proximal stomach but regardless of the locations, its specificity as a predictor of carcinoma is low. AIMS: The aim of this study was to establish whether human telomerase reverse transcriptase expression differs at various points in proximal versus distal gastric carcinogenesis and to test the utility of human telomerase reverse transcriptase expression as a marker of cancer risk in intestinal metaplasia. MATERIAL AND METHODS: Wax-embedded tissue from proximal and distal stomach including normal mucosa (n=86), intestinal metaplasia (n=83) and carcinoma (n=101) were used and slides were immunostained for human telomerase reverse transcriptase and pRb and scored semi-quantitatively. RESULTS: The results showed that in both proximal and distal stomach, human telomerase reverse transcriptase expression rates increased from normal mucosa to cancer. High rates of human telomerase reverse transcriptase expression were seen in the proliferative zones of glands in intestinal metaplasia. In both the locations, loss of pRb expression correlated with higher human telomerase reverse transcriptase expression. CONCLUSIONS: In conclusion, telomerase activity appears to be an early event in both proximal and distal gastric carcinogenesis and human telomerase reverse transcriptase is expressed in intestinal metaplasia. Telomerase re-expression may be facilitated by pRb inactivation. PMID- 15893284 TI - Prolonged prothrombin time, Factor VII and activated FVII levels in chronic liver disease are partly dependent on Factor VII gene polymorphisms. AB - BACKGROUND: Prothrombin time is a benchmark for functional assessment in cirrhosis and Factor VII levels (FVII), crucial in determining the prothrombin time, are genetically determined. METHODS: We have evaluated the prothrombin time, a number of haemostatic variables synthesised by the liver (FII, FV, FVII and activated FVII, AT and fibrinogen) and two polymorphisms of the FVII gene (5'F7 and 353R/Q) in: (a) patients with liver cirrhosis (n=118), (b) patients with chronic hepatitis (n=102) and (c) controls (n=100). RESULTS: By one-way analyses of variance, the prothrombin time and the mean levels of the FII, FV, FVIIc, FVIIa, and AT were statistically different between cirrhotics, chronic hepatitis patients and controls. The allele frequency of the FVII polymorphisms did not differ between the three groups. Those rare patients (4.6%) who were homozygous for the type 2 alleles had markedly reduced FVIIc and FVIIa levels. The analysis carried out taking into account Child class versus FVII genotype showed that the mean FVIIc levels were comparable for different genotypes within each Child's class, with the exception of the patients homozygous for the type 1 allele. CONCLUSION: Our findings help to explain the not infrequent finding of a severely prolonged prothrombin time in patients who are otherwise in a good functional class. PMID- 15893285 TI - Capsule endoscopy for the diagnosis and follow-up of blue rubber bleb nevus syndrome. AB - Blue rubber bleb nevus syndrome is a rare disorder characterised by cutaneous and gastrointestinal vascular malformations. A 31-year-old man came under our observation with obscure gastrointestinal bleeding. Hemangiomas on his back and the multiple bluish vascular lesions in the distal ileum revealed by capsule endoscopy led to the diagnosis of blue rubber bleb nevus syndrome. The patient was treated with octreotide to prevent rebleeding. During the 12 months of treatment he had no gastrointestinal bleeding episodes or anaemia. Then repeat capsule endoscopy showed that the small bowel vascular lesions were unchanged. Capsule endoscopy may be a useful tool for diagnosing and monitoring the effects of therapy in patients with blue rubber bleb nevus syndrome. PMID- 15893286 TI - Air swallowing can be responsible for non-response of heartburn to high-dose proton pump inhibitor. AB - Intraluminal electrical impedance is a novel technique, which is able for the first time to provide a qualitative assessment of refluxed material moving from the stomach to the oesophagus. In other words, the presence of air can be differentiated from that of liquid, because the former is characterised by high and the latter by low impedance compared with baseline. Moreover, the combined measurement of electrical impedance and pH-metry permits to distinguish acid from non-acid liquid reflux. One of the most important clinical applications of this method is to assess the reasons for poor response of GORD patients to high-dose proton pump inhibitors. This case report describes the results of impedance in the evaluation of a young woman, who did not respond to twice-daily doses of rabeprazole. She continued to complain of heartburn as major symptom and impedance allowed us to clarify that it was not related to acid or non-acid reflux, but to air swallowing. Therefore, this technique identified aerophagia to be responsible for persistent heartburn despite high-dose proton pump inhibitor and prevented the adoption of more aggressive, but probably unuseful therapies, such as the surgical one. PMID- 15893287 TI - Tricyclic antidepressants in the treatment of postoperative antireflux surgery symptoms. PMID- 15893288 TI - Low speed vehicle passenger ejection restraint effectiveness. AB - Current golf carts and LSV's (Low Speed Vehicles) produce a significant number of passenger ejections during sharp turns. These LSV's do not typically possess seatbelts, but do provide outboard bench seat hip restraints that also serve as handholds. However, many current restraint designs appear incapable of preventing passenger ejections due to their low height and inefficient handhold position. Alternative handhold and hip restraint designs may improve passenger safety. Accordingly, this paper examines minimum size requirements for hip restraints to prevent passenger ejection during sharp turns and evaluates the effectiveness of a handhold mounted at the center of the bench seat. In this study, a simulation of a turning cart supplies the dynamic input to a biomechanical model of an adult male seated in a golf cart. Various restraint combinations are considered, both with and without the central handhold, to determine the likelihood of passenger ejection. It is shown that only the largest restraint geometries prevent passenger ejection. Adequate hip restraints should be much larger than current designs and a central handhold should be provided. In this way, golf cart and LSV manufacturers could reduce passenger ejections and improve fleet safety by incorporating recommendations provided herein. PMID- 15893289 TI - Public health impact of drug resistant Plasmodium falciparum malaria. AB - The alarming increase in Plasmodium falciparum resistance to commonly used anti malarial drugs represents a major public health threat. The impact is however difficult to quantify. In low transmission areas, an increase in acute manifestations ("epidemic") is often quickly apparent and resistance is rapidly propagated due to high drug pressure on existing parasite populations. In high transmission areas, the clinical effects are mainly prolonged/chronic infections with increasing risk of severe anemia. Mortality estimates from public health records in Africa generally suggest significant increases (from 2- to 11-fold) in malaria-associated mortality among children when resistance develops and spreads. Hospital attendances and admissions show similar trends. Change of policy to alternative efficacious treatment with radical cure is necessary at an earlier stage (from 10% treatment failure) than previously assumed in order to prevent deaths in millions of African children. Early switch to artemisinin based combination therapy (ACT) represents such a critical and urgent strategy. PMID- 15893290 TI - Electromyography as a measure of peak and cumulative workload in intermediate care and its relationship to musculoskeletal injury: an exploratory ergonomic study. AB - Injury rates in Intermediate Care (IC) facilities are high and the factors related to these injuries are unclear. The objectives of this exploratory sub study, which is part of a large multi-faceted study in 8 IC facilities are to: (1) evaluate EMG measured over a full-shift in the back and shoulders of 32 care aides (CAs) as an indicator of peak and cumulative workload (n = 4 x 8 facilities); investigate the relationship between EMG measures and injury indicators; and explore the relationship between EMG measures and other workload measurements. Lumbar EMG was converted to predicted cumulative spinal compression and ranged in CAs from 11.7 to 22.8 MNs with a mean of 16.4 MNs. Average compression was significantly different during different periods of the day (p < 0.001) with highest compression during pre-breakfast when CAs assist most with activities of daily living. Significant differences were found in average compression between low and high injury facilities for 3 of 5 periods of the day (p < 0.010). Peak compressions exceeding 3400 N occurred for very little of the workday (e.g. 11.25s during the 75 min period pre-breakfast). Peak neck/shoulder muscle activity is low (99% APDF ranged from 8.33% to 28% MVC). Peak and cumulative spinal compression were significantly correlated with lost-time and musculoskeletal injury rates as well as with total tasks observed in the CAs (p < 0.01). Perceived exertion was only correlated with peak compressions (p < 0.01). Facilities with low injury rates provided significantly more CAs (p < 0.01) to meet resident needs, and subsequently CAs performed fewer tasks, resulting in less peak and cumulative spinal loading over the day. PMID- 15893291 TI - CO2 and O2 concentrations in integral motorcycle helmets. AB - Inhaling air which contains excess CO2 and/or is oxygen-deficient is known to present health risks and to diminish human cognitive abilities. The average CO2 concentrations relevant to a motorcyclist wearing an integral helmet were measured 20 years ago and found to be alarmingly large. The purpose of the present study was to examine gas concentrations typically inhaled by a motorcyclist. Average concentrations of CO2 near the upper lip for persons (n = 4) wearing integral motorcycle helmets were measured in the laboratory and the field to facilitate comparison to previous work, and similarly high average concentrations were found: above 2% when stationary, well below 1% for speeds of 50 km/h or more. Very good agreement was obtained between laboratory and field measurements. Detailed measurements of the time-dependent CO2 concentrations passing through a mouthpiece for mouth-breathing showed inhaled levels slightly over half of the corresponding average concentrations, including 1.3+/-0.3% at standstill, though higher concentrations (4% or more) were inhaled at the beginning of each breath. Opening the visor at standstill had on average no effect. At a speed of 50 km/h the inhaled CO2 concentration resembles that for a person without a helmet in still air, at about 0.2%. The oxygen deficiency is generally equal to the CO2 concentration, and could also contribute negatively to a motorcyclist's cognitive abilities. PMID- 15893292 TI - Functional studies with membrane-bound and detergent-solubilized alpha2 adrenergic receptors expressed in Sf9 cells. AB - A chip-based biosensor technology using surface plasmon resonance (SPR) was developed for studying the interaction of ligands and G protein-coupled receptors (GPCRs). GPCRs, the fourth largest superfamily in the human genome, are the largest class of targets for drug discovery. We have expressed the three subtypes of alpha(2)-adrenergic receptor (alpha(2)-AR), a prototypical GPCR as functional fusion proteins in baculovirus-infected insect cells. The localization of the expressed receptor was observed in intracellular organelles, as detected by eGFP fluorescence. In addition, the deletion mutants of alpha(2B)-AR, with a deletion in the 3rd intracellular loop, exhibited unaltered K(d) values and enhanced stability, thus making them more promising candidates for crystallization. SPR demonstrated that small molecule ligands can bind the detergent-solubilized receptor, thus proving that alpha(2)-AR is active even in a lipid-free environment. The K(d) values obtained from the biosensor analysis and traditional ligand binding studies correlate well with each other. This is the first demonstration of the binding of a small molecule to the detergent-solubilized state of alpha(2)-ARs and interaction of low-molecular mass-ligands in real time in a label-free environment. This technology will also allow the development of high throughput platform for screening a large number of compounds for generation of leads. PMID- 15893293 TI - Acute treatment of inpatients with psychotic symptoms using Acceptance and Commitment Therapy: pilot results. AB - Cognitive behavior therapy (CBT) has been demonstrated in a number of randomized controlled trials to be efficacious as an adjunctive treatment for psychotic disorders. Emerging evidence suggests the usefulness of CBT interventions that incorporate acceptance/mindfulness-based approaches for this population. The current study extended previous research by Bach and Hayes (2002. The use of Acceptance and Commitment Therapy to prevent the rehospitalization of psychotic patients: A randomized controlled trial. Journal of Consulting and Clinical Psychology, 70, 1129-1139) using Acceptance and Commitment Therapy (ACT) in the treatment of psychosis. Psychiatric inpatients with psychotic symptoms were randomly assigned to enhanced treatment as usual (ETAU) or ETAU plus individual sessions of ACT. At discharge from the hospital, results suggested short-term advantages in the ACT group in affective symptoms, overall improvement, social impairment, and distress associated with hallucinations. In addition, more participants in the ACT condition reached clinically significant symptom improvement at discharge. Although 4-month rehospitalization rates were lower in the ACT group, these differences did not reach statistical significance. Decreases in the believability of hallucinations during treatment were observed only in the ACT condition, and change in believability was strongly associated with change in distress after controlling for change in the frequency of hallucinations. Results are interpreted as largely consistent with the findings of Bach and Hayes and warrant further investigations with larger samples. PMID- 15893294 TI - Inhibition of dipeptidyl peptidase IV by novel inhibitors with pyrazolidine scaffold. AB - Inhibition of dipeptidyl peptidase IV (DPP-IV) activity has been reported to improve nutrient-stimulated insulin secretion through the stabilization of glucagon-like peptide (GLP-1). In the present study, we identified novel DPP-IV inhibitors of pyrazolidine derivatives (Compounds 1 and 2) and characterized their biological effects in vitro and in vivo. Compound 1, an isoleucine pyrazolidide with a phenyl urea group, inhibited rat plasma DPP-IV, porcine kidney DPP-IV, as well as human Caco-2 DPP-IV with IC(50) values of 1.70, 2.26, and 2.02 microM, respectively. Because of the poor pharmacokinetic properties of Compound 1, further optimization was carried out, leading to the discovery of Compound 2, which had similar in vitro activities. Compound 2 acted as a selective and competitive inhibitor of DPP-IV. MALDI-TOF mass spectrometric analysis proved that the compound (20 microM) effectively blocked the degradation of active GLP-1 peptide by 61%. Although similar in in vitro potency, marked improvement of in vivo efficacy and pharmacokinetic properties was seen with Compound 2. Oral administration of Compound 2 resulted in potent and rapid inhibition of circulating DPP-IV in C57BL/6J mice, with ED(50) values of 26mg/kg (s.c.) and 42mg/kg (p.o.). In addition, this compound improved glucose tolerance in ob/ob mice, as determined by an oral glucose tolerance test (OGTT). These results indicate that Compound 2 is a potent and selective DPP-IV inhibitor with oral anti-hyperglycemic activity in vivo. PMID- 15893295 TI - Estradiol administration mediates the inflammatory response to formalin in female rats. AB - Female rats demonstrate higher pain sensitivity than do males in various nociceptive assays of inflammation. In the present study, we found that estradiol (20%) replacement in ovariectomized rats attenuated the chronic phase of the formalin response but only at high formalin concentrations thought to rely on peripheral inflammation. An inactive isomer of estradiol, alpha-estradiol, failed to result in the same attenuation (P > 0.05). Our results suggest that estradiol's actions in inflammatory responses are mediated through genomic estrogen receptor-mediated mechanisms. PMID- 15893296 TI - Does conversion of ATP to adenosine terminate ATP-stimulated vasopressin release from hypothalamo-neurohypophyseal explants? AB - ATP stimulates vasopressin (VP) release from explants of the hypothalamo neurohypophyseal system (HNS), but the response is not sustained for the duration of exposure to ATP. Since adenosine, a metabolite of ATP, inhibits VP release from neurohypophysial terminals and adenosine receptors (AR) are expressed in supraoptic nucleus (SON) neurons, we postulated that conversion of ATP to adenosine contributed to termination of ATP-stimulated VP release from HNS explants. This was tested using a non-selective AR antagonist, 5-amino-9-chloro-2 (2-furyl)-1, 2, 4-triazolo [1, 5-c] quinazoline (CGS-15943). CGS-15943 did not affect basal VP release and did not alter the initial response to ATP. A selective A1R antagonist, 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX), increased basal VP release at 1 microM, without altering the response to ATP. However, at a higher concentration of DPCPX (10 microM), VP release was enhanced by ATP for an extended period of time. Inhibition of the enzymatic conversion of ATP to adenosine using a combination of a potent ecto-5'-nucleotidase inhibitor, alpha,beta-methylene adenosine 5'-diphosphate (AMP-CP), and a competitive substrate for ecto-5'-nucleotidase (guanosine monophosphate, GMP) did not affect basal VP release. Enzymatic inhibition did slightly prolong the response to ATP, but it was not sustained for the duration of exposure to ATP. We conclude that an endogenous inhibitory influence of adenosine decreases basal VP release from HNS explants and that conversion of exogenously applied ATP to adenosine contributes to termination of ATP-induced stimulation of VP release, but additional mechanisms such as receptor desensitization also limit the response to extended exposure to ATP. PMID- 15893297 TI - Concurrent nociceptive stimulation impairs the anxiolytic effect of midazolam injected into the periaqueductal gray in mice. AB - This study investigated whether the opportunity to avoid or escape the open arms of an elevated plus-maze (EPM) affects the antinociceptive response observed when mice are subjected to open arm confinement. Furthermore, in order to better characterize the relationship between emotion and antinociception in the EPM, we examined the behavioral effects of midazolam injection into the midbrain periaqueductal gray matter (PAG). As our main aim was to evaluate the relevance of different levels of approach-avoid conflict (i.e. the presence of open and closed arms) to maze-induced antinociception, mice were exposed to one of three types of EPM-a standard EPM (sEPM), an open EPM (oEPM: four open arms) or, as a control condition, an enclosed EPM (eEPM: four enclosed arms). Nociception was assessed using the formalin test. Twenty minutes after formalin injection (50 microl, 2.5% formalin) into the dorsal right hind paw, mice received an intra-PAG injection of saline or midazolam (10-20 nmol). Five minutes later, they were individually exposed to one of the mazes for 10 min (25-35 min after formalin injection). Videotapes of the test sessions were scored for a variety of behavioral measures including time spent licking the formalin-injected paw. To examine whether the effects of midazolam on anxiety-like behavior may have been influenced by concurrent nociceptive stimulation (i.e. formalin pretreatment), naive mice were submitted to a similar procedure to that described above for the sEPM test but without formalin pretreatment. Results showed that mice exposed to the oEPM spent significantly less time licking the injected paw compared to groups exposed to either the sEPM or eEPM. Although exposure to the sEPM induced anxiety-like behaviors (i.e. open arm avoidance), it did not result in antinociception. Intra-PAG infusions of midazolam failed to block oEPM-induced antinociception or to alter sEPM-induced anxiety in mice that had received formalin injection. However, under normal test conditions (i.e. in the absence of formalin-induced nociceptive stimulation), intra-PAG midazolam produced clear anti-anxiety effects in mice exposed to the sEPM. Findings are discussed in terms of different emotional states induced by the oEPM and sEPM and the influence of concurrent nociceptive stimulation on the anti-anxiety effect of intra-PAG midazolam. PMID- 15893298 TI - A novel accessory role of neutrophils in concanavalin A-induced hepatitis. AB - Concanavalin A (Con A)-induced hepatitis has been investigated as a model of T cell-mediated liver injury, in which IFN-gamma plays an essential role by inducing apoptosis of liver cells. Since a large number of neutrophils infiltrate into the liver in the model, the role of neutrophils was investigated in this study. Con A hardly caused liver injury in neutrophil-depleted mice, as assessed as to the plasma alanine aminotransferase level as well as histochemistry. Neutrophil-depleted mice also failed to produce IFN-gamma. Intracellular IFN gamma staining revealed that, among liver leukocytes, T and NK cells but not neutrophils are the main producers of IFN-gamma. Nylon wool-purified "T cells", however, failed to produce IFN-gamma in response to Con A in vitro, while the production was restored by the addition of neutrophils. Overall, this study suggests that neutrophils play a novel accessory role in IFN-gamma production in Con A-induced hepatitis. PMID- 15893299 TI - Determination of the tissue distribution and excretion by accelerator mass spectrometry of the nonadecapeptide 14C-Moli1901 in beagle dogs after intratracheal instillation. AB - Moli1901 is a 19 residue polycyclic peptide antibiotic which increases chloride transport and water mobilization in airway epithelium. These properties suggest that it may be a useful treatment for cystic fibrosis (CF). In this study, we used accelerator mass spectrometry (AMS) to quantify Moli1901 following administration of only 0.045 microCi of 14C-Moli1901 per dog. Limits of quantitation of AMS were 0.03 (urine) to 0.3 (feces) ng equiv. Moli1901/g. Administration of 14C-Moli1901 by intratracheal instillation (approximately 100 microg) into the left cranial lobe of the lung of beagle dogs resulted in retention of 64% of the dose in the left cranial lobe for up to 28 days. Whole blood and plasma concentrations of 14C were <5 ng/ml at all times after the dose. Concentrations of 14C in whole blood and plasma declined over the first day after the dose and rose thereafter, with the rise in plasma concentrations lagging behind those in whole blood. During the first 3 days after the dose, plasma accounted for the majority of 14C in whole blood, but after that time, plasma accounted for only 25-30% of the 14C in whole blood. Tissue (left and right caudal lung lobe, liver, kidney, spleen, brain) and bile concentrations were low, always less than 0.25% the concentrations found in the left cranial lung lobe. Approximately 13% of the dose was eliminated in urine and feces in 28 days, with fecal elimination accounting for about 10% of the dose. The data presented here are consistent with that obtained in other species. Moli1901 is slowly absorbed and excreted from the lung, and it does not accumulate in other tissues. Moli1901 is currently in the clinic and has proven to be safe in single dose studies in human volunteers and cystic fibrosis patients by the inhalation route. No information on the disposition of the compound in humans is available. This study in dogs demonstrates the feasibility of obtaining that information using 14C Moli1901 and AMS. PMID- 15893300 TI - Glucuronidation in therapeutic drug monitoring. AB - BACKGROUND: Glucuronidation is a major drug-metabolizing reaction in humans. A pharmacological effect of glucuronide metabolites is frequently neglected and the value of therapeutic drug monitoring has been questioned. However, this may not always be true. METHODS: In this review the impact of glucuronidation on therapeutic drug monitoring has been evaluated on the basis of a literature search and experience from the own laboratory. RESULTS: The potential role of monitoring glucuronide metabolite concentrations to optimize therapeutic outcome is addressed on the basis of selected examples of drugs which are metabolized to biologically active/reactive glucuronides. Furthermore indirect effects of glucuronide metabolites on parent drug pharmacokinetics are presented. In addition, factors that may modulate the disposition of these metabolites (e.g. genetic polymorphisms, disease processes, age, and drug-drug interactions) are briefly mentioned and their relevance for the clinical situation is critically discussed. CONCLUSION: Glucuronide metabolites can have indirect as well as direct pharmacological or toxicological effects. Although convincing evidence to support the introduction of glucuronide monitoring into clinical practice is currently missing, measurement of glucuronide concentrations may be advantageous in specific situations. If the glucuronide metabolite has an indirect effect on the pharmacokinetics of the parent compound, monitoring of the parent drug may be considered. Furthermore pharmacogenetic approaches considering uridine diphosphate (UDP) glucuronosyltransferases polymorphisms may become useful in the future to optimize therapy with drugs subject to glucuronidation. PMID- 15893301 TI - Clinical laboratory testing for the antiphospholipid syndrome. AB - The first aCL test was developed in 1983 and subsequently standardized. Although in the last 6 to 7 years, new and more specific tests have become available, the aCL ELISA and the LA tests are still the first choice to be used in diagnosis of APS. While there is now doubt that the anticardiolipin test is useful in the diagnosis of APS, limitations of the assay have caused uncertainty and misinterpretation of the value of the test. Utilization of validated ELISA kits with well-tested calibrators and an "in-house standard" may enable more reproducible measurements. Reporting results semiquantitatively preserves the clinical utilize of the test without the misinterpretation of a quantitative result that may lack precision. The development of newer tests such as the beta2GPI ELISA and the APhL ELISA Kit, utilizing the phospholipid mixture, give promise to a more specific and reliable diagnosis of APS, while retaining good sensitivity. Other tests such as ELISA for prothrombin antibodies and annexin V antibodies are still under development and will require standardization and extensive evaluation. The aCL test should continue to be done and included in the Sapporo criteria. The aCL test is not as specific as the anti-beta2GPI test, but it is very sensitive and together with the LA test should capture the majority of the APS patients. IgA aCL and anti-beta2GPI positivity alone is rare but occasionally found and shown to be associated with major clinical manifestations of APS. Therefore, it is now recommended to include both tests, IgA aCL and IgG, IgM and IgA anti-beta2GPI to confirm diagnosis of APS. PMID- 15893302 TI - Sox3 expression in undifferentiated spermatogonia is required for the progression of spermatogenesis. AB - Sox3, a member of the high mobility group (HMG) family of transcription factors, is expressed in neural progenitor cells and in the gonads. Targeted deletion of Sox3 in mice causes abnormal development of the diencephalon and Rathke's pouch, the progenitor of the anterior pituitary gland. Male and female mice are also infertile and exhibit a primary defect in gametogenesis. In this study, we examined the expression and function of Sox3 in C57BL/6 mice to better understand its role in spermatogenesis. Testis development was normal during embryogenesis. However, spermatogenesis failed to progress during the postnatal period, with germ cell loss beginning at postnatal day 10 (P10). By P14, Sox3 null mice were nearly agametic, retaining only Sertoli cells and undifferentiated spermatogonia. Pituitary gonadotropin and testosterone levels were normal, suggesting a defect in Sertoli cell and/or germ cell function. Immunostaining revealed that Sox3 was expressed in a subpopulation of germ cells localized at the base of the seminiferous tubules. Sox3 expression was restricted to proliferating germ cells and colocalized with neurogenin 3 (Ngn3), a helix-loop-helix transcription factor implicated in spermatogonial differentiation. The absence of Sox3 decreased Ngn3 and increased expression of Oct4, a marker of undifferentiated spermatogonia. We conclude that Sox3 is expressed in A(s), A(pr) and A(al) spermatogonia and is required for spermatogenesis through a pathway that involves Ngn3. PMID- 15893303 TI - Gene knockout analysis of two gamma-tubulin isoforms in mice. AB - Gamma-tubulin regulates the nucleation of microtubules, but knowledge of its functions in vivo is still fragmentary. Here, we report the identification of two closely related gamma-tubulin isoforms, TUBG1 and TUBG2, in mice, and the generation of TUBG1- and TUBG2-deficient mice. TUBG1 was expressed ubiquitously, whereas TUBG2 was primarily detected in the brain. The development of TUBG1 deficient (Tubg1-/-) embryos stopped at the morula/blastocyst stages due to a characteristic mitotic arrest: the mitotic spindle was highly disorganized, and disorganized spindles showed one or two pole-like foci of bundled MTs that were surrounded by condensed chromosomes. TUBG2 was expressed in blastocysts, but could not rescue the TUBG1 deficiency. By contrast, TUBG2-deficient (Tubg2-/-) mice were born, grew, and intercrossed normally. In the brain of wild-type mice, TUBG2 was expressed in approximately the same amount as TUBG1, but no histological abnormalities were found in the Tubg2-/- brain. These findings indicated that TUBG1 and TUBG2 are not functionally equivalent in vivo, that TUBG1 corresponds to conventional gamma-tubulin, and that TUBG2 may have some unidentified function in the brain. PMID- 15893304 TI - Foxg1 is required for specification of ventral telencephalon and region-specific regulation of dorsal telencephalic precursor proliferation and apoptosis. AB - Null mutation of the Foxg1 gene causes hypoplasia of the mouse telencephalon and loss of ventral telencephalic structures. We show that a crucial early requirement for Foxg1 is in the induction of ventral cell fate in the telencephalon. To study later proliferative defects, we have adapted an iododeoxyuridine and bromodeoxyuridine double labeling protocol for use in the developing embryo, which allows estimation of cell cycle kinetics in a single specimen. This technique is used to demonstrate that the cell cycle is prematurely lengthened in the Foxg1-null telencephalon. These defects are first apparent at embryonic day 10.5 (E10.5) and are most severe in the rostral telencephalon. We show that apoptosis is also reduced in the same rostral domain. These defects correspond temporally and spatially with a dramatic reduction in expression of the potent signaling molecule Fgf8. We also show that in the absence of Foxg1 an excess of neurons is produced from E11.5, depleting the progenitor pool and limiting the growth of the Foxg1(-/-) telencephalon. The increase in neurogenic division coincides with an increase in BMP signaling, as detected by immunohistochemistry for phosphorylated smad-1, -5, and -8. This study reinforces Foxg1's position as a major regulator of telencephalic neurogenesis and supports the idea that Foxg1 controls precursor proliferation via regulation of Fgf signaling and differentiation via regulation of Bmp signaling. PMID- 15893305 TI - Keeping an eye on the fly genome. AB - With its unique structure and dynamic development, the Drosophila eye has been a powerful genetic model system for studying molecular mechanisms of cell fate specification and differentiation. Hundreds of genes that function in a complex genetic network controlling this process have been identified during the past two decades. To further advance our understanding of the molecular mechanisms of eye development, it is increasingly important to place the current genetic pathway into a whole-genome perspective. Here, we review emerging technologies and strategies that will help to achieve this goal, including generation of a complete mutant set in Drosophila, genome-wide transcription factor target identification, and systematic studies of gene function aided by computational biology. PMID- 15893306 TI - Dynamic mitochondrial localization of nuclear transcription factor HMGA1. AB - It has been well established that high mobility group A1 (HMGA1) proteins act within the nucleus of mammalian cells as architectural transcription factors that regulate the expression of numerous genes. Here, however, we report on the unexpected cytoplasmic/mitochondrial localization of the HMGA1 proteins within multiple cell types. Indirect immunofluorescence, electron microscopic immunolocalization, and Western blot studies revealed that, in addition to the nucleus, HMGA1 proteins could also be found in both the cytoplasm and mitochondria of randomly dividing populations of wild-type murine NIH3T3 cells and transgenic human MCF-7 breast cancer epithelial cells expressing a hemagglutinin tagged-HMGA1a fusion protein. While the molecular mechanisms underlying these novel subcellular localization patterns have not yet been determined, initial synchronization studies revealed a dynamic, cell cycle dependent translocation of HMGA1 proteins from the nucleus into the cytoplasm and mitochondria of NIH3T3 cells. Furthermore, preliminary functionality studies utilizing a modified "chromatin" immunoprecipitation protocol revealed that HMGA1 retains its DNA binding capabilities within the mitochondria and associates with the regulatory D-loop region in vivo. We discuss potential new biological roles for the classically nuclear HMGA1 proteins with regard to the observed nucleocytoplasmic translocation, mitochondrial internalization, and regulatory D loop DNA binding. PMID- 15893307 TI - Retinoic acid induced myelomeningocele in fetal rats: characterization by histopathological analysis and magnetic resonance imaging. AB - The prevention of human neural tube defects by folic acid administration and the potential for fetal surgical intervention for myelomeningocele (MMC) have renewed interest in the molecular pathways and pathophysiology of spina bifida. Animal models for assessment of the early developmental biology and pathophysiology of this lesion are needed. The goal of this study was to develop and characterize a non-surgical rat model of MMC. Time-dated Sprague-Dawley rats were gavage fed different doses of retinoic acid (RA) dissolved in olive oil at E10 (maternal n = 55, fetal n = 505). Control animals received olive oil alone (maternal n = 20, fetal n = 265) or were untreated (maternal n = 5, fetal n = 63). Fetuses were analyzed by detailed histopathology and MRI. Overall, isolated MMC occurred in 60.7% (307/505) of RA-exposed fetuses and no controls. Histopathology confirmed the entire spectrum of severity observed in human MMC, ranging from exposure of the cord with intact neural elements to complete cord destruction. MRI of the brain of MMC fetuses confirmed structural changes similar to humans with Arnold Chiari malformation, including downward displacement of the cerebellum to just above the foramen magnum and compression of the developing medulla into a small posterior fossa. In conclusion, the RA-induced rat model of MMC is developmentally and anatomically analogous to human MMC. This relatively efficient and cost-effective model of MMC should facilitate investigation of the developmental biology and pathophysiology of MMC, and may be useful for the evaluation of further strategies for prenatal treatment. PMID- 15893308 TI - Direct, complex effects of estrogens on basal forebrain cholinergic neurons. AB - Although controversial, estrogens remain one of the few agents purported to influence the incidence of Alzheimer's disease and one of their postulated mechanisms of action is their effects on basal forebrain cholinergic neurons. However, it is unclear whether the responses of cholinergic neurons to estrogens are direct or mediated via the retrograde influences of neurotrophins, known to be induced by estrogens in the hippocampus and neocortex. In the present study, we explore the issue of the primary site of action of estrogens by studying the regulation of expression of genes that characterize mature cholinergic neurons, i.e., choline acetyltransferase, trkA, and p75(NTR) in the medial septum and the nucleus basalis complex. In parallel, we study the hippocampal expression of NGF, BDNF, and NT-3, i.e., neurotrophins with known trophic roles on cholinergic neurons. Gene expression is studied by RT-PCR in ovariectomized female rats with and without estrogen supplementation within the physiological estradiol range and in rats with complete fimbria-fornix transactions treated with estrogen or vehicle. To clarify mechanisms of estrogen transduction in cholinergic neurons, we study the effects of estrogen treatment on fimbria-fornix-lesioned mice with genetic ablations of ER subtypes alpha and beta. The results of the present study suggest that, while estrogens do regulate BDNF expression in the hippocampus and neocortex, they also exert stimulatory non-trophic effects on basal forebrain cholinergic neurons, primarily on ChAT expression. Cholinergic neurons retain their ability to respond to estrogens after their complete separation from the hippocampus. The elimination of ERalpha alters significantly the phenotypic responsiveness of cholinergic neurons to estrogens, whereas elimination of ERbeta appears to have no effect. Our findings support the idea that estrogens directly enhance cholinergic neuron function and that ERalpha plays a significant role in transducing these regulatory effects. PMID- 15893309 TI - Entamoeba histolytica EhPgp5 transcriptional activation depends on putative emetine response elements. AB - The multidrug resistance EhPgp5 gene promoter is active in drug resistant clone C2 trophozoites and its activity increases when trophozoites are cultured in the presence of emetine, suggesting that the EhPgp5 gene shows an inducible drug dependent mechanism. We analyzed different promoter fragments to detect those regions that activate transcription in the presence of emetine. Trophozoites were transfected with p375Pgp5, p259Pgp5, p187Pgp5, and p76Pgp5 plasmids and incubated with different emetine concentrations. p375Pgp5 and p259Pgp5 plasmids were able to drive CAT expression in A and C2 trophozoites only in the presence of emetine. CAT activity was turned off in the absence of drug. Interestingly, no CAT activity was detected in the presence or in the absence of emetine with p187Pgp5 plasmid in which 59 bp were deleted at the 5' end of the EhPgp5 minimal promoter (p259Pgp5). These results suggest that the overexpression of the EhPgp5 gene is a consequence of transcriptional activation of the gene promoter by putative drug responsive elements, located within the -111 to -170 bp of the transcription initiation site. PMID- 15893311 TI - Schistosoma mekongi: a prominent neutrophil chemotactic activity of egg antigen with reference to that of Schistosoma japonicum. AB - Schistosoma mekongi causes granulomatous lesions around eggs deposited in the liver with neutrophil-rich inflammatory reactions in the early stage of the egg laying. To define the aspects of the typical pathogenesis of S. mekongi infection, we determined the difference between soluble egg antigen (SEA) from S. mekongi and S. japonicum with a focus on chemotactic factors for neutrophils or eosinophils. Mean volume and protein amount of S. mekongi eggs was 71 and 58% of those of Schistosoma japonicum eggs, respectively. Neutrophil chemotactic activity of S. mekongi SEA was about two times higher than that of S. japonicum. In contrast, eosinophil chemotactic activity of S. mekongi SEA was about half of that of S. japonicum SEA. Molecular analysis revealed that S. mekongi SEA contains higher molecular-weight components with a lower level of glycosylation, and this is likely to be related to the intense neutrophil chemotactic activity in comparison with S. japonicum SEA. The prominent chemotactic reactivity for neutrophils is likely to be involved in the typical pathogenesis of mekongi schistosomiasis. PMID- 15893312 TI - 17-beta-estradiol activates maxi-K channels through a non-genomic pathway in human breast cancer cells. AB - We have investigated the acute effects of 17-beta-estradiol (E2) on K+ channels in MCF-7 breast epithelial cancer cells. E2 induced a rapid and irreversible augmentation of the K+ current for all membrane potentials superior to -25 mV. The effect of E2 was sensitive to Iberiotoxin, Charybdotoxin and TEA and can be elicited in the presence of the anti-estrogen ICI 182780 or be mimicked by the membrane impermeant form E2/BSA. Furthermore, E2/BSA was able to stimulate cell proliferation in a maxi-K inhibitors-sensitive manner. Thus, these results permit us to identify the maxi-K channel as the molecular target of E2 that regulates cell proliferation independently of the estrogen receptor. PMID- 15893313 TI - The inhibitory mechanism of curcumin and its derivative against beta-catenin/Tcf signaling. AB - We investigated the inhibitory mechanism of curcumin and its derivative (CHC007) against beta-catenin/T-cell factor (Tcf) signaling in various cancer cell lines. Curcumin is known to inhibit beta-catenin/Tcf transcriptional activity in HCT116 cells but not in SW620 cells. To clarify the inhibitory effect of curcumin against beta-catenin/Tcf signaling, we tested several cancer cell lines. In addition, in order to verify the inhibitory mechanism, we performed reporter gene assay, Western blot, immunoprecipitation, and electrophoretic mobility shift assay. Since inhibitors downregulated the transcriptional activity of beta catenin/Tcf in HEK293 cells transiently transfected with S33Y mutant beta-catenin gene, whose product is not induced to be degraded by adenomatous polyposis coli Axin-glycogen synthase kinase 3beta complex, we concluded that the inhibitory mechanism was related to beta-catenin itself or downstream components. Western blot analysis suggested that no change in the amount of cytosolic and membranous beta-catenin in a cell occurred; however, nuclear beta-catenin and Tcf-4 proteins were markedly reduced by inhibitors and this lead to the diminished association of beta-catenin with Tcf-4 and to the reduced binding to the consensus DNA. In the present study, we demonstrate that curcumin and its derivative are excellent inhibitors of beta-catenin/Tcf signaling in all tested cancer cell lines and the reduced beta-catenin/Tcf transcriptional activity is due to the decreased nuclear beta-catenin and Tcf-4. PMID- 15893314 TI - Structure of the ATPase subunit CysA of the putative sulfate ATP-binding cassette (ABC) transporter from Alicyclobacillus acidocaldarius. AB - CysA, the ATPase subunit of a putative sulfate ATP-binding cassette transport system of the gram-positive thermoacidophilic bacterium Alicyclobacillus acidocaldarius, was structurally characterized at a resolution of 2.0 Angstroms in the absence of nucleotides. In line with previous findings on ABC-ATPases the structures of the two monomers (called CysA-1 and CysA-2) in the asymmetric unit differ substantially in the arrangement of their individual (sub)domains. CysA-2 was found as a physiological dimer composed of two crystallographically related monomers that are arranged in an open state. Interestingly, while the regulatory domain of CysA-2 packs against its opposing domain that of CysA-1 undergoes a conformational change and, in the dimer, would interfere with the opposing monomer thereby preventing solute translocation. Whether this conformational state is used for regulatory purposes will be discussed. PMID- 15893315 TI - Wobble modification deficiency in mutant tRNAs in patients with mitochondrial diseases. AB - Point mutations in mitochondrial (mt) tRNA genes are associated with a variety of human mitochondrial diseases. We have shown previously that mt tRNA(Leu(UUR)) with a MELAS A3243G mutation and mt tRNA(Lys) with a MERRF A8344G mutation derived from HeLa background cybrid cells are deficient in normal taurine containing modifications [taum(5)(s(2))U; 5-taurinomethyl-(2-thio)uridine] at the anticodon wobble position in both cases. The wobble modification deficiency results in defective translation. We report here wobble modification deficiencies of mutant mt tRNAs from cybrid cells with different nuclear backgrounds, as well as from patient tissues. These findings demonstrate the generality of the wobble modification deficiency in mutant tRNAs in MELAS and MERRF. PMID- 15893316 TI - The complete set of tRNA species in Nanoarchaeum equitans. AB - The archaeal parasite Nanoarchaeum equitans was found to generate five tRNA species via a unique process requiring the assembly of seperate 5' and 3' tRNA halves [Randau, L., Munch, R., Hohn, M.J., Jahn, D. and Soll, D. (2005) Nanoarchaeum equitans creates functional tRNAs from separate genes for their 5'- and 3'-halves. Nature 433, 537-541]. Biochemical evidence was missing for one of the computationally-predicted, joined tRNAs designated as tRNA(Trp). Our RT-PCR and sequencing results identify this tRNA as tRNA(Lys) (CUU) joined at the alternative position between bases 30 and 31. We show that the intron-containing tRNA(Trp) was misidentified in the initial Nanoarchaeum equitans genome annotation [E. Waters et al. (2003) The genome of Nanoarchaeum equitans: insights into early archaeal evolution and derived parasitism. Proc. Natl. Acad. Sci. USA 100, 12984-12988]. Along with a previously unidentified joined tRNA(Gln) (UUG), Nanoarchaeum equitans exhibits 44 tRNAs and is enabled to read all 61 sense codons. Features unique to this set of tRNA molecules are discussed. PMID- 15893317 TI - Exploring blocker binding to a homology model of the open hERG K+ channel using docking and molecular dynamics methods. AB - Binding of blockers to the human voltage-gated hERG potassium channel is studied using a combination of homology modelling, automated docking calculations and molecular dynamics simulations, where binding affinities are evaluated using the linear interaction energy method. A homology model was constructed based on the available crystal structure of the bacterial KvAP channel and the affinities of a series of sertindole analogues predicted using this model. The calculations reproduce the relative binding affinities of these compounds very well and indicate that both polar interactions near the intracellular opening of the selectivity filter as well as hydrophobic complementarity in the region around F656 are important for blocker binding. These results are consistent with recent alanine scanning mutation experiments on the blocking of the hERG channel by other compounds. PMID- 15893318 TI - Reconciling tensions between research, evidence-based practice and user participation: time for nursing to take the lead. PMID- 15893319 TI - Contrasting effects of acute and chronic gastro-intestinal helminth infections on a heterologous immune response in a transgenic adoptive transfer model. AB - We have previously found that co-immunisation with ovalbumin (OVA) and the body fluid of the helminth Ascaris suum inhibited an OVA-specific delayed type hypersensitivity (DTH) response by reducing OVA-specific CD4+ T lymphocyte proliferation via an IL-4 independent mechanism. In the present study, we determined whether parasite infections themselves could induce similar changes to peripheral immunisation by examining the modulation of OVA-specific immune responses during acute and chronic helminth infections. Surprisingly, an acute infection with Trichinella spiralis, but not a chronic infection with Heligmosomoides polygyrus, inhibited the OVA-specific DTH reaction. Correspondingly, the T helper 1 (Th1) OVA-specific response was decreased in mice infected with T. spiralis, but not with H. polygyrus. Inhibition of the Th1 response may be a result of a shift in the Th1/Th2 balance as although both H. polygyrus and T. spiralis infected mice induced a Th2 OVA-specific response, that exhibited by T. spiralis was more potent. Furthermore, although IL-10 secretion upon OVA restimulation was similarly increased by both infections, production of this immunoregulatory cytokine may play a role in the suppression of immune responses observed with T. spiralis infection depending on the context of its release. Interestingly, analysis of the OVA-specific T lymphocyte division by carboxyfluorescein diacetate succinimidyl ester (CFSE) staining revealed that gastro-intestinal infection with the acute helminth T. spiralis, but not with chronic H. polygyrus, inhibited the systemic immune response by significantly inhibiting the antigen-specific T cell proliferation during the primary response, a mechanism similar to that observed when A. suum parasite extracts were directly mixed with the OVA during immunisation in our previous studies. PMID- 15893320 TI - Associations between IgG antibody to oral organisms and carotid intima-medial thickness in community-dwelling adults. AB - AIMS: The aims of this study are to describe the relationships between IgG antibodies to 17 oral organisms and atherosclerosis as indexed by carotid intima medial wall thickness (IMT) and to evaluate the role of smoking. METHODS AND RESULTS: Our study is based on a subset of participants in the Atherosclerosis Risk in Communities (ARIC) Study, who received a complete periodontal examination during visit 4 (1996-1998). The outcome was mean carotid IMT>/=1mm assessed by B mode ultrasound. The exposures were serum IgG antibody levels against 17 periodontal organisms using a whole bacterial checkerboard immunoblotting technique. Evaluation of all 17 antibodies indicated that antibody to Campylobacter rectus resulted in the best-fitting model (OR=2.3, 95% CI=1.83 2.84) and individuals with both high C. rectus and Peptostreptococcus micros titers had almost twice the prevalence of IMT>/=1mm than those with only a high C. rectus antibody (8.3% versus 16.3%). Stratification by smoking indicated that all microbial models significant for smokers were also significant for never smokers except for Porphyromonas gingivalis (p=0.08). CONCLUSIONS: This is the first study to report a relationship between IgG antibody reactive to oral organisms and subclinical atherosclerosis with significant relationships evident in both ever and never smokers. PMID- 15893321 TI - Development of a culture system that supports adult microglial cell proliferation and maintenance in the resting state. AB - Microglial cells constitute what is considered to be a fixed macrophage population in the central nervous system (CNS), which are broadly implicated in the regulation of neuroinflammation. In the normal adult CNS, microglial cells exist in a resting state characterized by a minimal or negative expression of MHC class II and the co-stimulatory molecules CD80, CD86 and CD40 and exhibit a unique ramified morphology. Microglial cell activation is associated with many inflammatory and neurogenerative CNS pathologies and is characterized by the transformation of resting microglia into cells with a macrophage morphology and up-regulation of MHC class II and co-stimulatory molecules. The cellular and molecular mechanisms required for microglial cell activation and their immunological functions in the adult brain still remain enigmatic, primarily due to the lack of an appropriate culture system that both facilitates microglial survival and expansion in the resting state. Here, we describe a new M-CSF dependent culture system that overcomes these barriers and allows the long-term proliferation and maintenance of resting adult microglial cells isolated from the CNS. These cultured microglial cells retain their plasticity as indicated by their ability to up-regulate MHC class II and differentiate into cells with a macrophage morphology following the addition of IFN-gamma and GM-CSF, or activated T cells, which produce both cytokines. By measuring the proliferation of the T cells, we were also able to demonstrate that the microglial cells differentiated into fully functional antigen presenting cells. In addition, the replacement of the M-CSF with GM-CSF resulted in the differentiation of microglial cells into cells morphologically and phenotypically similar to dendritic cells. Our microglial cell culture system is the first described that allows the expansion of adult cells in the resting state and will facilitate studies examining the specific mechanisms of microglial cell activation and functions involved in a variety of CNS pathologies. PMID- 15893322 TI - Time-dependent changes in myocardial structure following discrete injury in mice deficient of matrix metalloproteinase-3. AB - Myocardial scars from radiofrequency (RF) ablation can increase in size in the post-injury period, resulting in remodeling of the extracellular matrix (ECM). The matrix metalloproteinases (MMPs) contribute to adverse myocardial remodeling following injury. However, the role of specific MMP types in RF scar enlargement remains unclear. One MMP type, MMP-3, degrades a wide range of ECM substrates and can activate other MMPs. This project examined LV remodeling in wild type (WT) and MMP-3 deficient (mmp-3-/-) mice following RF injury. RF lesions (0.5 mm probe, 80 degrees C, 30 s) were created on the LV epicardium of WT (C57/BL6) and mmp-3-/- mice and were terminally studied at 1 h, 3, 7, and 28 days post-RF (n=10 each). Heart mass indexed to tibial length (mg/mm) was similar in the WT and mmp 3-/- mice at 1 h (8.1+/-0.3 vs. 7.6+/-0.3), but lower in the mmp-3-/- mice at 28 days post-RF (11.9+/-0.4 vs. 10.5+/-0.4, P<0.05). Scar volumes were greater in the mmp-3-/- mice at 3 days, but similar in the two groups at 28 days. Immunohistochemical localization showed fewer macrophages and lymphocytes at the scar border at 3 days in the mmp-3-/- hearts, but similar staining for these cells in WT and mmp-3-/- hearts at 7 and 28 days post-RF. Post-RF, the early increase in scar volume was accelerated in mmp-3-/- mice and associated with abnormal inflammatory cell infiltration/migration to the area of injury. These findings define a mechanistic role for MMP-3 in RF scar expansion and provide a temporal window during which interruption of MMP-3 activation may impair post-RF myocardial wound healing. PMID- 15893323 TI - Differential nucleotide regulation of KATP channels by SUR1 and SUR2A. AB - ATP-sensitive potassium (K(ATP)) channels are heterooctamers of an inward rectifier potassium channel (Kir6) and a sulfonylurea receptor (SUR, a member of the ATP-binding cassette (ABC) transporter family). In the pancreatic beta-cell K(ATP) channels are dynamically active, and transgenic expression of overactive Kir6.2 mutants leads to severe neonatal diabetes and death, while in the ventricular cardiomyocyte they are closed except under conditions of severe metabolic inhibition, and similarly overactive transgenes are without gross phenotypic consequence. This discrepancy may arise in part from differences at the molecular level between the two SUR isotypes that constitute the regulatory subunit of the K(ATP) channel in those tissues: SUR1 in the pancreas, SUR2A in the heart. K(ATP) channels generated from coexpression of Kir6.2 with SUR1 exhibit greater MgADP stimulation than channels generated from coexpression of Kir6.2 with SUR2A. This difference persists when the open state stability of the channel is enhanced by application of PIP(2), consistent with each isotype transducing an intrinsically different energetic contribution to the channel pore. When expressed as isolated, affinity-purified protein constructs, NBF2 of SUR1 exhibits increased in vitro ATP hydrolysis compared to NBF2 of SUR2A. This biochemical difference may underlie the increased MgADP stimulation exhibited by SUR1-containing channels vs. SUR2A-containing channels, which may in turn contribute to physiological differences, observed at the tissue level, between pancreatic and cardiac K(ATP) channels. PMID- 15893324 TI - Conformational states and thermodynamics of alpha-lactalbumin bound to membranes: a case study of the effects of pH, calcium, lipid membrane curvature and charge. AB - The study of the conformational changes of bovine alpha-lactalbumin, switching from soluble states to membrane-bound states, deepens our knowledge of the behaviour of amphitropic proteins. The binding and the membrane-bound conformations of alpha-lactalbumin are highly sensitive to environmental factors, like calcium and proton concentrations, curvature and charge of the lipid membrane. The interactions between the protein and the membrane result from a combination of hydrophobic and electrostatic interactions and the respective weights of these interactions depend on the physicochemical conditions. As inferred by macroscopic as well as residue-level methods, the conformations of the membrane-bound protein range from native-like to molten globule-like states. However, the regions anchoring the protein to the membrane are similar and restricted to amphiphilic alpha-helices. H/(2)H-exchange experiments also yield residue-level data that constitute comprehensive information providing a new point of view on the thermodynamics of the interactions between the protein and the membrane. PMID- 15893325 TI - Desolvation is a likely origin of robust enthalpic barriers to protein folding. AB - Experimental data from global analyses of temperature (T) and denaturant dependence of the folding rates of small proteins led to an intrinsic enthalpic folding barrier hypothesis: to a good approximation, the T-dependence of folding rate under constant native stability conditions is Arrhenius. Furthermore, for a given protein, the slope of isostability folding rate versus 1/T is essentially independent of native stability. This hypothesis implies a simple relationship between chevron and Eyring plots of folding that is easily discernible when both sets of rates are expressed as functions of native stability. Using experimental data in the literature, we verify the predicted chevron-Eyring relationship for 14 proteins and determine their intrinsic enthalpic folding barriers, which vary approximately from 15 kcal/mol to 40 kcal/mol for different proteins. These enthalpic barriers do not appear to correlate with folding rates, but they exhibit correlation with equilibrium unfolding enthalpy at room temperature. Intrinsic enthalpic barriers with similarly high magnitudes apply as well to at least two cases of peptide-peptide and peptide-protein association, suggesting that these barriers are a hallmark of certain general and fundamental kinetic processes during folding and binding. Using a class of explicit-chain C(alpha) protein models with constant elementary enthalpic desolvation barriers between C(alpha) positions, we show that small microscopic pairwise desolvation barriers, which are a direct consequence of the particulate nature of water, can act cooperatively to give rise to a significant overall enthalpic barrier to folding. This theoretical finding provides a physical rationalization for the high intrinsic enthalpic barriers in protein folding energetics. Ramifications of entropy-enthalpy compensation in hydrophobic association for the height of enthalpic desolvation barrier are discussed. PMID- 15893326 TI - Identification of amino acid residues in the human immunodeficiency virus type-1 reverse transcriptase tryptophan-repeat motif that are required for subunit interaction using infectious virions. AB - The human immunodeficiency virus type-1 (HIV-1) reverse transcriptase (RT) functions as a heterodimer (p51/p66), which makes disruption of subunit interactions a possible target for antiviral drug design. Our understanding of subunit interface interactions has been limited by the lack of virus-based approaches for studying the heterodimer. Therefore, we developed a novel subunit specific mutagenesis approach that enables precise molecular analysis of the heterodimer in the context of infectious HIV-1 particles. Here, we analyzed the contributions of amino acid residues comprising the Trp-motif to RT subunit interaction and function. Our results reveal important inter- and intra-subunit interactions of residues in the Trp-motif. A tryptophan cluster in p51 (W398, W402, W406, W414), proximal to the interface, was found to be important for p51/p66 interaction and stability. At the dimer interface, residues W401, Y405 and N363 in p51 and W410 in p66 mediate inter-subunit interactions. The W401 residue is critical for RT dimerization, exerting distinct effects in p51 and p66. Our analysis of the RT heterodimerization enhancing non-nucleoside RT inhibitor (NNRTI), efavirenz, indicates that the effects of drugs on RT dimer stability can be examined in human cells. Thus, we provide the first description of subunit-specific molecular interactions that affect RT heterodimer function and virus infection in vivo. Moreover, with heightened interest in novel RT inhibitors that affect dimerization, we demonstrate the ability to assess the effects of RT inhibitors on subunit interactions in a physiologically relevant context. PMID- 15893327 TI - Screening for anxiety disorders in depressed patients. AB - The Psychiatric Diagnostic Screening Questionnaire (PDSQ) is a brief, psychometrically strong, questionnaire designed to screen for common Axis I disorders. In the present report, we examine the ability of the PDSQ to identify anxiety disorders in psychiatric outpatients with a principal diagnosis of major depressive disorder. Eight hundred patients presenting for treatment were evaluated with the Structured Clinical Interview for DSM-IV (SCID) after completing the PDSQ. Two hundred ninety-five patients had a principal diagnosis of major depressive disorder. The mean sensitivity and negative predictive value of the anxiety disorder subscales was 88.5% and 96.5% when all patients were considered, and 88.2% and 95.6% when only depressed patients were examined. The PDSQ's anxiety disorder subscales have high sensitivity and negative predictive value thereby indicating that the scale could function well as a screening instrument in depressed patients. PMID- 15893328 TI - Cortical lesions and brain atrophy in MS. AB - Multiple sclerosis is generally considered a disease of the white matter. However, this is only one pathological aspect of the disease as demyelination is prominent in the grey matter of deep cerebral nuclei and the cerebral cortex. In this review, we discuss the possibility that disease involvement of grey matter structures may significantly contribute to clinical disability in multiple sclerosis patients. PMID- 15893329 TI - Multiple calmodulin mRNAs are selectively transported to functionally different neuronal and glial compartments in the rat hippocampus. An electron microscopic in situ hybridization study. AB - The ultrastructural distribution of the calmodulin (CaM) mRNAs transcribed from the three CaM genes was studied in the CA1 region of the adult rat hippocampus by means of electron microscopic in situ hybridization. Digoxigenin-labeled CaM gene specific riboprobes were detected with nanogold-anti-digoxigenin antibody conjugate. The CaM mRNAs were differentially distributed in both the neuronal and glial cell compartments. The greatest difference in neuronal distribution of the CaM mRNAs was found in the dendrites, where the mRNAs transcribed from the CaM I and III genes were much more abundant than the CaM II mRNA. The neuronal perikarya were heavy labeled for all the CaM mRNAs. Interestingly, the myelinated axons and axon terminals also contained small amounts of nanogold particles for all the CaM mRNAs, which diminished with increasing distance from the soma. Most of the synaptic profiles, however, contained labeling only in the postsynaptic region. The CaM mRNAs were differentially distributed in the glial cells. While the glial cell somata were only lightly labeled, surprisingly concentrated labeling was present in the perisynaptic and perivascular astrocytic processes. In general, the CaM II mRNA was the least represented in the glial processes. Only a very low CaM gene expression was observed in the endothelial and resting microglial cells. These results provide ultrastructural evidence for differential targeting of the multiple CaM mRNA transcripts to the intracellular compartments and suggest their microdomain-specific regulation. PMID- 15893330 TI - Accumulation of organochlorines and brominated flame retardants in estuarine and marine food chains: field measurements and model calculations. AB - Food chain accumulation of organochlorines and brominated flame retardants in estuarine and marine environments is compared to model estimations and fresh water field data. The food chain consists of herbivores, detritivores and primary and secondary carnivores i.e. fish, fish-eating birds and marine mammals. Accumulation of polychlorinated biphenyls is predicted well by OMEGA for herbi detritivores and primary and secondary carnivorous fish. Ratios are similar to those found for fresh water species. Accumulation ratios for fish-eating birds and mammals are overestimated by the model, which is attributed partly to biotransformation of meta-para unsubstituted congeners. Additionally, birds may feed in other less polluted areas. For brominated diphenylethers (BDE) accumulation patterns are highly species and congener specific. Accumulation depends on both K(ow) and metabolization capacities. BDE47 is the predominant congener in lower trophic levels. For marine birds and mammals accumulation ratios of BDE99 and 100 are similar to or higher than ratios of persistent PCBs. PMID- 15893331 TI - Mercury distribution in Douro estuary (Portugal). AB - Determinations of dissolved reactive and total dissolved mercury, particulate and sedimentary mercury, dissolved organic carbon (DOC), particulate organic carbon (POC) and suspended particulate matter (SPM) have been made in the estuary of river Douro, in northern Portugal. The estuary was stratified by salinity along most of its length, it had low concentrations of SPM, typically <20 mg dm(-3), and concentrations of DOC in the range <1.0-1.8 mg dm(-3). The surface waters had a maximum dissolved concentration of reactive mercury of about 10 ng dm(-3), whereas for the more saline bottom waters it was about 65 ng dm(-3). The surface waters had maximum concentrations of total suspended particulate mercury of approximately 7 microg g(-1) and the bottom waters were always <1 microg g(-1). Concentrations of mercury in sediments was low and in the range from 0.06 to 0.18 microg g(-1). The transport of mercury in surface waters was mainly associated with organic-rich particulate matter, while in bottom waters the dissolved phase transport of mercury is more important. Lower particulate organic matter, formation of chlorocomplexes in more saline waters and eventually the presence of colloids appear to explain the difference of mercury partitioning in Douro estuarine waters. PMID- 15893332 TI - First assessment of the Caulerpa racemosa (Caulerpales, Chlorophyta) invasion along the French Mediterranean coast. AB - The introduced green alga Caulerpa racemosa var. cylindracea has been rapidly spreading in the Mediterranean Sea since 1990. It was first observed in France in 1997 (Marseilles). In early 2004, the stretch of the French Mediterranean coastline and the surface area affected by the invasion were estimated at about 83 km and 4014 ha, respectively. The depth range of colonized areas was usually 10-35 m depth. Shallow (0-10 m) and deep (down to 40 m) dense meadows were rarely observed. In contrast to the dead matte of Posidonia oceanica, which constituted the most widely colonized substratum, dense P. oceanica meadows and fine sand with large ripple-marks were not invaded. Few rocky areas were colonized and coarse sand bottoms were usually colonized below 20 m depth. All the colonized areas were exposed to human activities and more than 40% were fishing areas. Mild climate, suitable substrata, presence of vectors of dispersal and absence of efficient biological control make the French Mediterranean coast particularly vulnerable to the further spread of the alga. PMID- 15893333 TI - Induction, adaptation and recovery of biological responses: implications for environmental monitoring. AB - A wide range of biological responses have been used to identify exposure to contaminants, monitor spatial and temporal changes in contamination levels, provide early warning of environmental deterioration and indicate occurrences of adverse ecological consequences. To be useful in environmental monitoring, a biological response must reflect the environmental stress over time in a quantitative way. We here argue that the time required for initial induction, maximum induction, adaptation and recovery of these stress responses must first be fully understood and considered before they can be used in environmental monitoring, or else erroneous conclusions (both false-negative and false positive) may be drawn when interpreting results. In this study, data on initial induction, maximum induction, adaptation and recovery of stress responses at various biological hierarchies (i.e., molecular, biochemical, physiological, behavioral, cytological, population and community responses) upon exposure to environmentally relevant levels of contaminants (i.e., metals, oil, polycyclic aromatic hydrocarbons (PAHs), organochlorines, organophosphates, endocrine disruptors) were extracted from 922 papers in the biomarker literature and analyzed. Statistical analyses showed that: (a) many stress responses may decline with time after induction (i.e., adaptation), even if the level of stress remains constant; (b) times for maximum induction and recovery of biochemical responses are positively related; (c) there is no evidence to support the general belief that time for induction of responses at a lower biological hierarchy (i.e., molecular responses and biochemical responses) is shorter than that at higher hierarchy (i.e., physiological, cytological and behavioral responses), although longer recovery time is found for population and community responses; (d) there are significant differences in times required for induction and adaptation of biological responses caused by different types of contaminants; (e) times required for initial and maximum induction of physiological responses in fish are significantly longer than those in crustaceans; and (f) there is a paucity of data on adaptation and recovery of responses, especially those at population and community levels. The above analyses highlight: (1) the limitations and possible erroneous conclusions in the present use of biomarkers in biomonitoring programs, (2) the importance of understanding the details of temporal changes of biological responses before employing them in environmental management, and (3) the suitability of using specific animal groups as bioindicator species. PMID- 15893334 TI - Intermittent exposure to reduced oxygen levels affects prey size selection and consumption in swimming crab Thalamita danae Stimpson. AB - Portunid crabs Thalamita danae (carapace width: 46-56 mm) were exposed to low oxygen level (4.0 mg O2 l(-1)) and hypoxia (1.5 mg O2 l(-1)) for 6 h each day with three size classes (large: 15.0-19.9 mm, medium: 10.0-14.9 mm, small: 5.0 9.9 mm) of mussels Brachidontes variabilis offered as food. Consumption rate, prey size preference, and prey handling including breaking time, handling time, eating time and prey value, were studied during the time the crabs were exposed to reduced oxygen levels and results were compared with the crabs maintained at high oxygen level (8.0 mg O2 l(-1)) throughout the experiment. Consumption of mussels from all size classes was significantly higher at high oxygen level than at reduced oxygen levels. No mussel size preference was observed for crabs exposed to 4.0 or 8.0 mg O2 l(-1) but those crabs exposed to 1.5 mg O2 l(-1) preferred medium mussels. Both breaking time and handling time increased with mussel size but did not vary with oxygen level. Prey value of each mussel consumed (mg dry wt eaten crab(-1) s(-1)) was calculated by dividing the estimated dry weight of the mussel by the observed handling time. Mean prey value varied significantly with mussel size, with values obtained for large mussels being higher than small mussels at 4.0 and 8.0 mg O2 l(-1); the effect of oxygen level, however, was insignificant. In view of portunid crabs as major predators of mussels, results may help explain dominance of mussels in eutrophic harbours in Hong Kong. PMID- 15893335 TI - Chlorinated pesticides and PCBs in the sea-surface microlayer and seawater samples of Singapore. AB - Sea-surface microlayer (SML) and seawater samples collected from Singapore's coastal marine environment were analyzed for selected chlorinated pesticides and polychlorinated biphenyls (PCBs). The SML is a potential site of enrichment of persistent organic pollutants (POPs) compared to the underlying water column. The concentration ranges of SigmaHCH, SigmaDDT and SigmaPCB in subsurface (1 m depth) seawater were 0.4-27.2 ng/l (mean 4.0 ng/l), 0.01-0.6 ng/l (mean 0.1 ng/l) and 0.05-1.8 ng/l (mean 0.5 ng/l) respectively. In the SML, the concentration ranges of SigmaHCH, SigmaDDT and SigmaPCB were 0.6-64.6 ng/l (mean 9.9 ng/l), 0.01-0.7 ng/l (mean 0.2 ng/l) and 0.07-12.4 ng/l (mean 1.3 ng/l) respectively. High spatial and temporal distribution was observed for all POPs measured. However, overall levels measured in the SML were lower than levels reported in the literature for SML samples from temperate coastal regions-possibly due to loss of semi-volatile compounds in the tropical climate of Singapore. Atmospheric wet deposition during the monsoon season may be an important source of POPs to the SML. This study provides the first scientific data on POP concentrations and enrichment factors in the SML for Southeast Asia. PMID- 15893336 TI - On statistical tests of phylogenetic tree imbalance: the Sackin and other indices revisited. AB - We investigate the distribution of statistical measures of tree imbalance in large phylogenies. More specifically, we study normalized versions of the Sackin's index and the number of subtrees of given sizes. Using the connection with structures from theoretical computer science, we provide precise description for the limiting distribution under the null hypothesis of Yule trees. Corrected p-values are then computed, and the statistical power of these statistics for testing the Yule model against a model of biased speciation is evaluated from simulations. As an illustration, the tests are applied to the HIV-1 reconstructed phylogeny. PMID- 15893337 TI - Accuracy in the estimation of quantitative minimal area from the diversity/area curve. AB - The problem of representativity is fundamental in ecological studies. A qualitative minimal area that gives a good representation of species pool [C.M. Bouderesque, Methodes d'etude qualitative et quantitative du benthos (en particulier du phytobenthos), Tethys 3(1) (1971) 79] can be discerned from a quantitative minimal area which reflects the structural complexity of community [F.X. Niell, Sobre la biologia de Ascophyllum nosodum (L.) Le Jolis en Galicia, Invest. Pesq. 43 (1979) 501]. This suggests that the populational diversity can be considered as the value of the horizontal asymptote corresponding to the curve sample diversity/biomass [F.X. Niell, Les applications de l'index de Shannon a l'etude de la vegetation interdidale, Soc. Phycol. Fr. Bull. 19 (1974) 238]. In this study we develop a expression to determine minimal areas and use it to obtain certain information about the community structure based on diversity/area curve graphs. This expression is based on the functional relationship between the expected value of the diversity and the sample size used to estimate it. In order to establish the quality of the estimation process, we obtained the confidence intervals as a particularization of the functional (h-phi)-entropies proposed in [M. Salicru, M.L. Menendez, D. Morales, L. Pardo, Asymptotic distribution of (h,phi)-entropies, Commun. Stat. (Theory Methods) 22 (7) (1993) 2015]. As an example used to demonstrate the possibilities of this method, and only for illustrative purposes, data about a study on the rocky intertidal seawed populations in the Ria of Vigo (N.W. Spain) are analyzed [F.X. Niell, Estudios sobre la estructura, dinamica y produccion del Fitobentos intermareal (Facies rocosa) de la Ria de Vigo. Ph.D. Mem. University of Barcelona, Barcelona, 1979]. PMID- 15893338 TI - Reinforcement learning for a stochastic automaton modelling predation in stationary model-mimic environments. AB - In this paper we propose a mathematical learning model for the feeding behaviour of a specialist predator operating in a random environment occupied by two types of prey, palatable mimics and unpalatable models, and a generalist predator with additional alternative prey at its disposal. A well known linear reinforcement learning algorithm and its special cases are considered for updating the probabilities of the two actions, eat prey or ignore prey. Each action elicits a probabilistic response from the environment that can be favorable or unfavourable. To assess the performance of the predator a payoff function is constructed that captures the energetic benefit from consuming acceptable prey, the energetic cost from consuming unacceptable prey, and lost benefit from ignoring acceptable prey. Conditions for an improving predator payoff are also explicitly formulated. PMID- 15893339 TI - SVM classification of human intergenic and gene sequences. AB - Despite constant improvement in prediction accuracy, gene-finding programs are still unable to provide automatic gene discovery with the desired correctness. This paper presents an analysis of gene and intergenic sequences from the point of view of language analysis, where gene and intergenic regions are regarded as two different subjects written in the four-letter alphabet {A,C,G,T}, and high frequency simple sequences are taken as keywords. A measurement alpha(l(tau)) was introduced to describe the relative repeat ratio of simple sequences. Threshold values were found for keyword selections. After eliminating 'noise', 178 short sequences were selected as keywords. DNA sequences are mapped to 178-dimensional Euclidean space, and SVM was used for prediction of gene regions. We showed by cross-validation that the program we developed could predict 93% of gene sequences with 7% false positives. When tested on a long genomic multi-gene sequence, our method improved nucleotide level specificity by 21%, and over 60% of predicted genes corresponded to actual genes. PMID- 15893340 TI - CXCR4 and CCR5 regulation and expression patterns on T- and monocyte-macrophage cell lineages: implications for susceptibility to infection by HIV-1. AB - Chemokine receptor expression may vary dramatically among cell subsets. Therefore, the stage of differentiation and the lineage of CD4 cells may profoundly affect their susceptibility to infection by human immunodeficiency virus type 1 (HIV-1). However, the mechanisms of coreceptor competition for association with HIV-1 glycoproteins remain unknown. Here, we propose mathematical models that address the interdependence of the concentrations of CD4 and CCR5 for efficient infection by M-tropic HIV-1 as well as additional complications originated by coreceptor competition caused by posttranslational modifications that positively or negatively affect the coreceptor ability to form complexes with CD4 and/or HIV-1 envelope. Furthermore, since CCR5 and CXCR4 expression on human leukocytes designate these cells as HIV-1 potential targets, the expression of the major HIV-1 coreceptors are also dynamically modeled/quantified as function of the stage of cell differentiation. Results show that although coreceptor competition degree has limited influence on R5 strain infectivity, the infectivity of CXCR4-using isolates strongly depends on the CD4 expression, according to the coreceptor competition model proposed in Lee et al. [J. Virol. 74(11) (2000) 5016]. Understanding the role of in vivo alterations in CD4, CCR5 and CXCR4 densities on HIV-1 cell entry may help the development of optimal control strategies for AIDS pathogenesis. PMID- 15893341 TI - Optimization of harvesting age in an integral age-dependent model of population dynamics. AB - The paper deals with optimal control in a linear integral age-dependent model of population dynamics. A problem for maximizing the harvesting return on a finite time horizon is formulated and analyzed. The optimal controls are the harvesting age and the rate of population removal by harvesting. The gradient and necessary condition for an extremum are derived. A qualitative analysis of the problem is provided. The model shows the presence of a zero-investment period. A preliminary asymptotic analysis indicates possible turnpike properties of the optimal harvesting age. Biological interpretation of all results is provided. PMID- 15893342 TI - Change in mitochondrial membrane potential is the key mechanism in early warm hepatic ischemia-reperfusion injury. AB - The mitochondrion has been proposed to be both a target and a perpetuator of hepatic ischemia-reperfusion (IR) injury because of its reactive oxygen species (ROS) formation. Our hypothesis is that subcellular derangement in mitochondrial function is one of the earliest steps leading to the early IR-mediated loss of hepatocellular integrity. Under chloralhydrate anesthesia (36 mg/kg BW), Sprague Dawley rats (n=7) were subjected to 40 min of warm hepatic lobular ischemia followed by 60 min reperfusion. Rats (n=7) without hepatic IR were used as controls. The fluorochromes rhodamine 123 and bisbenzimide were administered intravenously for observation of changes in mitochondrial membrane potential and hepatocellular viability, respectively. Intravital fluorescence microscopy (IVFM) was performed prior to ischemia and at 15, 45, and 60 min after reperfusion in the experimental group and at corresponding time points in the control group. A parallel relationship between mitochondrial membrane potential and cell viability as reflected in a concomitant reduction in nuclear and cytoplasmic fluorescence intensity during IR was demonstrated (r2=0.76, P<0.05). The diminution in fluorescence intensities also correlated significantly with the elevation in plasma transaminase activities (r2>0.90, P<0.05). Our data suggested that alteration in mitochondrial membrane potential is a critical subcellular event leading to hepatocellular damage in the early phase of hepatic IR injury. PMID- 15893343 TI - Rapid response learning in amnesia: delineating associative learning components in repetition priming. AB - Functional brain imaging studies of priming assume that the behavioral facilitation and activity reductions resulting from multiple repetitions reflect the continued tuning of processes engaged during the initial processing of items. Utilizing an object priming paradigm in which participants were asked to make relative size judgments about visually presented common objects, we tested an alternate hypothesis that states that with multiple repetitions participants come to rely on a more efficient response learning mechanism. In experiment 1, the decision cue was inverted such that previous judgments made either once or three times were rendered invalid. Decision inversion resulted in a reduction of all priming, but most critically, led to a reduction of multiple-repetition priming to the level of single-repetition priming. In experiment 2, patients with amnesia failed to show a priming advantage for multiple repetitions, indicating that response learning is dependent on the medial temporal lobes. Taken together, these results suggest that a different process increasingly mediates priming behavior as repetitions increase. With repeated exposure, behavioral facilitation rapidly comes to reflect a more efficient response learning mechanism rather than facilitated access to object knowledge. PMID- 15893344 TI - Potassium intake during cell dehydration. AB - Isotonic NaCl is ingested in addition to water by cell-dehydrated rats in two bottle tests. The objective of the present work was to find out whether mineral intake in the cell-dehydrated rat is specific to NaCl in a five-bottle test. Adult male Sprague Dawley rats had distilled water and four mineral solutions at palatable concentrations (0.01 M KCl, 0.05 mM CaCl2, 0.15 M NaHCO3, 0.15 M NaCl) simultaneously available for consumption. Cell-dehydration was produced infusing 1.5 ml of NaCl solution (0.15, 0.25, 0.5, 1.0, 2.0, 4.0 M) intravenously for 10 min and intakes were recorded for the next hour. It was observed a NaCl concentration-dependent increase in 0.01 M KCl intake. The ingestion of the other mineral solutions was not significantly altered compared to infusion of 0.15 M NaCl. The ingestion of KCl was not related to changes in serum potassium concentration. The ingestion of KCl was reduced in half and water was the preferred fluid when the five-bottle test was performed with mineral solutions at isomolar (0.15 M) concentrations. There was no increase in intake of other mineral solution in the isomolar test. No preference was observed for palatable or isomolar solutions during early extracellular dehydration until 4 h after subcutaneous injection of furosemide, in spite of the increase in total volume intake. Therefore, mineral intake induced by cell dehydration is not specific for NaCl solution. The type of mineral solution available influences the choice and KCl is the preferred solution of the cell-dehydrated rat in the conditions of the present study. PMID- 15893346 TI - Primary care, social inequalities and all-cause, heart disease and cancer mortality in US counties: a comparison between urban and non-urban areas. AB - OBJECTIVE: The objective of this study was to test whether the association between primary care and income inequality on all-cause, heart disease and cancer mortality at county level differs in urban (Metropolitan Statistical Area-MSA) compared with non-urban (non-MSA) areas. STUDY DESIGN: The study consisted of a cross-sectional analysis of county-level data stratified by MSA and non-MSA areas in 1990. Dependent variables included age and sex-standardized (per 100,000) all cause, heart disease and cancer mortality. Independent variables included primary care resources, income inequality, education levels, unemployment, racial/ethnic composition and income levels. METHODS: One-way analysis of variance and multivariate ordinary least squares regression were employed for each health outcome. RESULTS: Among non-MSA counties, those in the highest income inequality category experienced 11% higher all-cause mortality, 9% higher heart disease mortality, and 9% higher cancer mortality than counties in the lowest income inequality quartile, while controlling for other health determinants. Non-MSA counties with higher primary care experienced 2% lower all-cause mortality, 4% lower heart disease mortality, and 3% lower cancer mortality than non-MSA counties with lower primary care. MSA counties with median levels of income inequality experienced approximately 6% higher all-cause mortality, 7% higher heart disease mortality, and 7% higher cancer mortality than counties in the lowest income inequality quartile. MSA counties with low primary care (less than 75th percentile) had significantly lower levels of all-cause, heart disease and cancer mortality than those counties with high primary care. CONCLUSIONS: In non MSA counties, increasing primary physician supply could be one way to address the health needs of rural populations. In MSA counties, the association between primary care and health outcomes appears to be more complex and is likely to require intervention that focuses on multiple fronts. PMID- 15893347 TI - Smoking behaviour, cessation attempts and the influence of parental smoking in older adult women: a cross-sectional analysis from Italy. AB - OBJECTIVES: To investigate the association between parental smoking during infancy/adolescence and smoking prevalence in older adult women, and to provide a description of smoking and smoking cessation patterns in this subset of the population. STUDY DESIGN: Cross-sectional survey. METHODS: Between 1999 and 2001, trained physicians in 11 health agencies throughout the Abruzzo Region, Italy, conducted semi-structured interviews on 9708 women aged 50-70 years attending mammographic screening (overall response rate 89%). Information was collected on sociodemographic characteristics, smoking habits, parental smoking during their infancy and/or adolescence, and exposure to environmental tobacco smoke (ETS) inside or outside the family. RESULTS AND CONCLUSIONS: The overall prevalence of current smoking was 15.6% (n=1516), and that of former smoking was 13.2%. Women whose parent(s) smoked were more likely to be current smokers, especially if only the mother smoked (adjusted odds ratio 4.27; 95% confidence interval 2.24-8.12). Other factors significantly associated with current smoking in the multivariate analysis were younger age, lower body mass index, higher level of education, unmarried status, and exposure to ETS either inside or outside the family environment. Eighteen percent of all current smokers were non-inhalers, and more than 60% of ex-smokers quit smoking on their first attempt. Our findings expand the existing evidence suggesting that a strong effect of parental conduct on a daughter's smoking behaviour may persist throughout life. Although more research is needed, especially to clarify the role of genetic and environmental factors in determining the mother-child smoking association, our results suggest that intensifying smoking prevention efforts directed at women with children might considerably reduce the risk of ever smoking in future female generations. PMID- 15893348 TI - Quality and variability of osteoporosis data in general practice computer records: implications for disease registers. AB - OBJECTIVE: To determine the extent to which routinely collected general practitioner computer data could be used to create disease registers of patients with osteoporosis, and to report any improvement in data quality since previous studies. STUDY DESIGN: Audit using anonymized data extracted from general practice computer records from across England. METHODS: Morbidity Query Information and Export Syntax (MIQUEST) software was used to extract structured data from the 78 volunteer practices that participated in the study. The data were aggregated and analysed. RESULTS: There were 100-fold differences in the rates of recording of relevant data. Many patients receiving treatment had no diagnostic codes. Data about secondary causes of osteoporosis and fractures were more consistently recorded than data relating to falls. There were no data to indicate whether fractures were low impact. T-scores, the gold-standard measure of bone density, were very infrequently recorded. CONCLUSIONS: Sufficient data about secondary causes of osteoporosis exist, and these could be searched to identify patients at risk. Meanwhile, fracture recoding could be improved, including likely fragility fractures, and T-scores could be added to computer records. A systematic approach is needed to raise the computer records to a standard where they can be used as valid and reliable disease registers. PMID- 15893349 TI - A study of seven different types of grafts for jugular vein transplantation in the horse. AB - The aim of this study was to investigate the biological behaviour of vascular grafts replacing a section of the jugular vein in order to improve the results of the surgical treatment of complete thrombosis of the jugular vein in the horse. Seven graft types: fresh allograft, home frozen allograft, glutaraldehyde-fixed allograft, cryo-preserved allograft, PTFE-graft (Gore), small intestinal submucosa preparation (Cook) and fresh autograft, were randomly implanted in ponies. The grafts were removed after one month and examined histologically for: preservation of the graft structures, acceptance by the host, intima proliferation, presence of endothelium and patency. The glutaraldehyde- and cryopreserved grafts show reasonable results and the PTFE and autograft had the best results especially with respect to host acceptance, endothelium presence and patency. Further research is necessary to improve graft behaviour, especially to the aspect of endothelisation. Obstruction of the jugular vein in horses can be treated surgically. PMID- 15893350 TI - Effect of antineoplastic drugs on the expression of Bcl-2 and Bcl-xL genes in the feline T-cell leukemia cell line. AB - The Bcl-2 gene is the first member of a rapidly expanding family of genes that regulate apoptosis. Bcl-2 has been shown to repress cell death triggered by a diverse array of stimuli including chemotherapy and gamma-irradiation. Chemotherapy of feline lymphoma is generally carried out with antineoplastic drugs, which are reported to induce apoptosis in tumor cells. However, the precise apoptotic signals, induced by chemotherapeutic drugs against feline tumors have not been fully characterized. Therefore, we have evaluated the expression of Bcl-2 and Bcl-xL in FT-1 upon in vitro treatment with these drugs. In the present study, full length of feline Bcl-xL gene was sequenced, and the expressions of Bcl-2 and Bcl-xL mRNAs in feline lymphoma cell line (FT-1) cultured with doxorubicin, prednisolone or vincristine were investigated. Feline Bcl-xL clone was 1163 base pairs in length and encoded 233 amino acids. The predicted amino acid sequence was 99.1%, 98.7%, 96.1%, 97.4%, 97.0% and 97.9% homologous to predicted Bcl-xL of dog, human, mouse, pig, rat and sheep, respectively. The levels of Bcl-2 transcripts at 24h incubation in FT-1 stimulated with doxorubicin (0.3mug/ml), prednisolone (0.2mug/ml) and vincristine (5ng/ml) were increased to about 41.0-, 62.0- and 11.1-fold to those in non stimulated FT-1, respectively. On the other hand, the level of Bcl-xL transcripts at 24h incubation in FT-1 stimulated by doxorubicin and prednisolone were significantly increased about 4.2- and 5.8-folds to the controls and inducible level of Bcl-xL by vincristine was decreased about 0.35-folds. PMID- 15893351 TI - Failure of a killed Leishmania amazonensis vaccine against American cutaneous leishmaniasis in Colombia. AB - We report the results of a double-blind, randomized, placebo-controlled clinical trial of a killed whole-cell Leishmania amazonensis candidate vaccine against American cutaneous leishmaniasis (CL) in Colombia. The trial subjects were 2597 healthy volunteers with negative leishmanin skin test (LST) selected from rural Colombian soldiers who were going to patrol endemic areas. They were randomized to receive either three doses of vaccine (n=1295) or placebo (n=1302) given at 20 day intervals. An active and passive case detection system was established to follow-up volunteers for 1 year after vaccination. Safety and efficacy of the vaccine were determined by comparing local and systemic adverse reactions after each dose and the incidence of parasitologically confirmed CL. In the vaccine and placebo groups 101 (7.7%) and 88 (6.8%) volunteers developed CL respectively. The vaccine was shown to be safe but offered no protection against CL caused by L. panamensis in the proposed vaccination schedule. PMID- 15893352 TI - Low serum allopregnanolone levels in girls with precocious pubarche. AB - Allopregnanolone, a neuroactive steroid, increases during pubertal development and high concentrations are present in subjects with precocious puberty. The aim of the present study was to evaluate serum allopregnanolone levels in girls with precocious pubarche (PP). Basal gonadotropins and steroid hormones were assessed in 17 girls with PP, 22 girls with central precocious puberty (CPP), 25 girls with normal puberty at the same pubertal stage of CPP ones, and 17 prepubertal girls. Adrenocorticotropin hormone (ACTH) and gonadotropin-releasing hormone (GnRH) stimulation tests were performed in all subjects with PP, and in 12 out of 22 with CPP. All girls with normal puberty underwent to GnRH test, while ACTH test was performed in 17 out of 25. Basal dehydroepiandrosterone sulfate (DHEAS) concentrations resulted significantly higher in PP and normal pubertal girls than in prepubertal ones. Allopregnanolone, gonadotropins and estradiol levels were significantly lower in PP group with respect to CPP (P<0.05), while they were comparable among PP, normal pubertal and prepubertal groups. After ACTH administration, allopregnanolone concentrations significantly increased in all groups (P<0.05). After GnRH stimulation, its levels significantly increased in CPP and normal pubertal controls (P<0.05), while no incremental rise was found in PP girls. In conclusion, our study shows that in girls with PP basal and GnRH stimulated levels of allopregnanolone are significantly lower than in CPP girls. These data suggest that this neurosteroid may be considered a new marker of pubertal development. PMID- 15893353 TI - A rat model of smoke inhalation injury: influence of combustion smoke on gene expression in the brain. AB - Acute smoke inhalation causes death and injury in victims of home and industrial fires as well as victims of combat situations. The lethal factors in combustion smoke inhalation are toxic gases and oxygen deficiency, with carbon monoxide (CO) as a primary cause of death. In survivors, inhalation of smoke can result in severe immediate and delayed neuropathologies. To gain insight into the progression of molecular events contributing to smoke inhalation sequelae in the brain, we developed a smoke inhalation rat model and conducted a genome-wide analysis of gene expression. Microarray analysis revealed a modified brain transcriptome with changes peaking at 24 h and subsiding within 7 days post smoke. Overall, smoke inhalation down regulated genes associated with synaptic function, neurotransmission, and neurotrophic support, and upregulated genes associated with stress responses, including nitric oxide synthesis, antioxidant defenses, proteolysis, inflammatory response, and glial activation. Notably, among the affected genes, many have been previously implicated in other types of brain injury, demonstrating the usefulness of microarrays for analysis of changes in gene expression in complex insults. In accord with previously described modulations of nitric oxide homeostasis in CO poisoning, microarray analysis revealed increased brain expression of nitric oxide synthase (NOS) and NOS ligand after inhalation of smoke. Furthermore, immunostaining showed significant elevations in perivascular NOS and in protein nitration, corroborating the involvement of nitric oxide perturbations in post-smoke sequelae in the brain. Thus, the new rat model, in combination with microarray analyses, affords insight into the complex molecular pathophysiology of smoke inhalation in the brain. PMID- 15893355 TI - Rapid protection of cattle from direct challenge with foot-and-mouth disease virus (FMDV) by a single inoculation with an adenovirus-vectored FMDV subunit vaccine. AB - We have previously demonstrated that swine vaccinated with one dose of a replication-defective human adenovirus type 5 (Ad5) vector containing the capsid and 3C proteinase coding regions of foot-and-mouth disease virus (FMDV) were protected when challenged 7 days later with homologous virus. In the current study, we have extended this approach to cattle, the most economically important animals susceptible to FMD. Five cattle were vaccinated with the Ad5-FMDV subunit vaccine and these animals and 2 co-housed control animals were challenged intradermolingually 7 days later. Both control animals developed typical signs of FMD including fever and vesicular lesions on all 4 feet. All 5 vaccinated animals were protected against disseminated disease. PMID- 15893356 TI - Kinetics of cyanogen chloride destruction by chemical reduction methods. AB - In this study, membrane introduction mass spectrometry (MIMS) was applied to evaluate the kinetics of cyanogen chloride (ClCN) destruction by chemical reduction methods, using thiosulfate, sulfite, metabisulfite, ferrous ions and zero-valent iron at various concentrations and pH. The ClCN destruction followed second-order reaction kinetics in all cases of using sulfur compounds, though the second-order rate constants varied substantially from approximately 0.3-25.7 M( 1)s(-1) under different experimental conditions. The destruction of ClCN was primarily attributable to the chemical reduction pathway. Hydroxide-assisted ClCN hydrolysis was only significant at pH 9 and also when the observed reduction rate was relatively slow. The second-order rate constants achieved by sulfur(IV) compounds in the form of sulfite were found to be higher than those obtained with thiosulfate and S(IV) compounds in the form of bisulfite. Ferrous ions and zero valent iron demonstrated slow or no ClCN reduction up to dosages of 1000 mgL(-1) and 100 gL(-1), respectively. These findings suggest that applying moderately high dosages of S(IV) compounds under neutral or alkali conditions with sufficient contact time is required for wastewater ClCN destruction. In addition, ClCN losses during long-term preservation with excess reducing sulfur compounds prior to analysis can be substantial and should be avoided. PMID- 15893357 TI - Effect of humic acid on the photolysis of the pesticide atrazine in a surfactant aided soil-washing system in acidic condition. AB - The photolytic destruction of Atrazine (ATZ) following a surfactant-aided soil washing process was investigated in the presence of humic acid (HA). A non-ionic surfactant, Brij 35, was found to be a good solving agent, extracting ATZ without causing any retardation effect on the photolysis process. However, the HA that was co-extracted from the surfactant-aided soil-washing system was found to be capable of improving the photolysis of ATZ at low concentrations, while quenching the photodegradation of ATZ at higher concentrations. By considering the light attenuation effect due to surfactant and HA, the quantum yields of the system with respect to the proposed reaction mechanisms of the associated excited states of ATZ were investigated and modeled. The relative kinetic rates of the dominant reaction mechanisms (i.e., deactivation of triplet, direct photolysis of triplet, product formation, and quenching of triplet) were then compared. PMID- 15893358 TI - Bioconcentration factor of relatively low concentrations of chlorophenols in Japanese medaka. AB - Bioconcentration factors (BCF) for pentachlorophenol (PCP) and 2,4-dichlorophenol (2,4-DCP) in Japanese medaka (Oryzias latipes) were determined at five different concentrations of the chemicals, between 0.1 and 10 microg/l (PCP), 0.3 and 30 microg/l (2,4-DCP), in the ambient water. Medaka were exposed to each chemicals in a continuous-flow system during the embryonic development period and 60 days after hatching from eggs collected in the laboratory. Both the exposure time and the aqueous concentrations are much more realistic and closer to natural aquatic environments than those used in conventional BCF studies. The BCF values of PCP were from (4.9+/-2.8)x10(3) at the aqueous concentration of 0.074+/-0.028 microg/l to (2.1+/-1.4)x10(3) at 9.70+/-0.56 microg/l. The BCF value of 2,4-DCP were from (3.4+/-3.0)x10(2) at 0.235+/-0.060 microg/l to 92+/-27 at 27.3+/-1.6 microg/l. Generally, BCF values increased as the aqueous concentrations of PCP or 2,4-DCP decreased. This finding suggests that a relatively low and realistic aqueous concentration of these compounds is necessary to more accurately determine their BCF values in natural aquatic environments. Conventional BCF experiments at higher aqueous concentrations may underestimate the BCF values. PMID- 15893359 TI - Determination of the dietary habits of a Magdalenian woman from Saint-Germain-la Riviere in southwestern France using stable isotopes. AB - To obtain direct dietary information, carbon and nitrogen isotope ratios were measured from bone collagen acquired from the well-preserved skeleton of a Magdalenian woman from the site of Saint-Germain-la-Riviere in southwestern France. Comparison of delta13C and delta15N values of the human bone collagen to those of bone collagen from local herbivores and carnivores indicates that the woman's primary source of protein was the meat of large terrestrial herbivores. Application of a linear mixing model to the woman's isotopic signature indicates that (1) no significant marine-derived protein contributed to her average diet; (2) saiga antelope, which dominates the faunal remains at Saint-Germain-la Riviere, was not the main source of terrestrial protein; and (3) her pattern of subsistence reflects a less opportunistic behavior than generally attributed to humans from this period. Dietary proportions of prey reflected by the number of identified specimens are revised using meat percentage estimates, which de emphasize the importance of saiga antelope in human subsistence at Saint-Germain la-Riviere during the middle Magdalenian. PMID- 15893360 TI - cDNA representational difference analysis used in the identification of genes related to the aging process in rat kidney. AB - Aging is a complex physiological process by which the functions of many organ systems deteriorate. Growing evidence shows that age-related changes and damage are causally related to oxidative stress and inflammatory responses from reactive species. The aim of this study was to identify differentially expressed genes in old and young kidneys of Fisher 344 male rats during the aging process using complementary DNA representational difference analysis (cDNA RDA). cDNA RDA is a subtractive technique for identifying a focused set of differentially expressed genes. The distinctive advantage of this technique is its capability of detecting differences in gene expressions at less than one copy per cell and identifying genes not previously described in the database. Reverse transcription-polymerase chain reaction with specific primers was applied to confirm the differences found by RDA. Twenty-one putative differentially expressed genes were identified. Sixteen genes were up-regulated during aging and were associated with stress response and inflammatory reactions, while five genes were down-regulated. These data suggested that the inflammatory process is a plausible cause of the aging process. PMID- 15893361 TI - Membrane alteration as a basis of aging and the protective effects of calorie restriction. AB - As has been experimentally determined, oxidative modification to biological systems can be extensive, although the identification and stochiometric relation of the reactive species that cause these alterations have not been fully elucidated. In this review, arguments are presented to support the notion that the combined effects of membrane lipid peroxidation and its by-products, reactive aldehydes are likely responsible for membrane-associated functional declines during aging. As evidence for a systemic response to overall oxidative stress, the molecular inflammation hypothesis of aging is discussed by considering that the activation of inflammatory genes act as a bridge linking normal aging to pathological processes. PMID- 15893362 TI - Examination of the requirement for ucp-4, a putative homolog of mammalian uncoupling proteins, for stress tolerance and longevity in C. elegans. AB - Reactive oxygen species (ROS) are generated by mitochondrial respiration and can react with and damage cellular components. According to the free radical theory of aging, oxidative damage from mitochondrial ROS is a major cause of cellular decline during aging. Mitochondrial uncoupling proteins (UCPs) uncouple ATP production from electron transport and can be stimulated by free radicals, suggesting UCPs may perform a cytoprotective function. The nematode, Caenorhabditis elegans, contains one UCP-like protein, encoded by the ucp-4 gene. We have investigated the genetic requirement for ucp-4 in normal aging and stress resistance. Consistent with the hypothesis that ucp-4 encodes a putative uncoupling protein, animals lacking ucp-4 function contained elevated ATP levels. However, the absence of ucp-4 function did not affect adult lifespan or survival in the presence of thermal or oxidative stress. Together, these results demonstrate that ucp-4 is a negative regulator of ATP production in C. elegans, but is not required for normal lifespan. PMID- 15893363 TI - Toward a unified theory of caloric restriction and longevity regulation. AB - The diet known as calorie restriction (CR) is the most reproducible way to extend the lifespan of mammals. Many of the early hypotheses to explain this effect were based on it being a passive alteration in metabolism. Yet, recent data from yeast, worms, flies, and mammals support the idea that CR is not simply a passive effect but an active, highly conserved stress response that evolved early in life's history to increase an organism's chance of surviving adversity. This perspective updates the evidence for and against the various hypotheses of CR, and concludes that many of them can be synthesized into a single, unifying hypothesis. This has important implications for how we might develop novel medicines that can harness these newly discovered innate mechanisms of disease resistance and survival. PMID- 15893364 TI - Metsulfuron spray drift reduces fruit yield of hawthorn (Crataegus monogyna L.). AB - This study was carried out to investigate whether spray drift of metsulfuron has a potential to negatively affect hawthorn (Crataegus monogyna) hedgerows near agricultural fields. For this purpose four doses of metsulfuron ranging from 5% to 40% of the field dose (4 g metsulfuron per hectare) were sprayed on trees in seven different hawthorn hedgerows. The actual deposition on the leaves was measured by means of a tracer (glycine). Spraying was conducted both at the bud stage and at early flowering. Leaves, flowers, green berries and mature berries were harvested and the number and weight of each were measured. The spraying at the bud stage caused a highly significant reduction in number and dry weight of berries, whereas it had no effects on leaf and flower production. The berry reduction was close to 100% at actual depositions relevant for spray drift under normal conditions. Spraying at early flowering also significantly reduced berries although the effect was smaller than for the spraying at bud stage. The early flower stage spraying caused no reduction in number and size of leaves. The possible ecological consequence is that metsulfuron spray drift from agricultural fields has a potential to reduce the amount of berries available for frugivorous birds in nearby hedgerows. A potential need for regulatory measures to reduce herbicide spray drift to hedgerows situated near agricultural fields with herbicide use is also indicated. PMID- 15893365 TI - Evaluation and prediction of emissions from a road built with bottom ash from municipal solid waste incineration (MSWI). AB - In autumn 2001, a full-scale test road was built with municipal solid waste incineration (MSWI) bottom ash at the Davamyran landfill, Umea, Northern Sweden. Leachates were collected from asphalted sections with either bottom ash or gravel as filling material. In this research, 12 months of ash leachate sampling were evaluated with respect to emissions of contaminants such as trace metals and chlorides (Cl). The usefulness of regression models describing trace metal mobility from bottom ash was also tested as predictive tools for reusability applications of MSWI bottom ash. Cl, Cu, and Cr had the highest mobility (considering leachate concentrations) in the ash leachate, though concentrations of Cl and Cu decreased during the sampling period (Cl from 10,000 to 600 mg l( 1); Cu from 1600 to 500 microg l(-1)). An increased mobility of Cr during the autumns (about 3-4 times higher compared to the summer) was noted with a maximum value of nearly 70 microg l(-1) during autumn 2001. Pb showed a very low mobility over the entire year with leachate concentrations of around 3-4 microg l(-1). Chemical equilibrium calculations using Minteq indicated that several Cu minerals were oversaturated in the leachate, thus mineral precipitation could be responsible for declining amounts of Cu in the leachate. Adsorption to iron oxides was found to be a probable explanation for the low mobility of Pb. A reasonably good agreement between regression models and field values were achieved for Ni, Pb, Zn, and Cu, while the models for Cd and Cr were less promising. Even though a large part of the variation (R2=61-97%) in the leaching experiment could be explained by only pH and L/S, field data were much more scattered than expected from field pH. PMID- 15893366 TI - Relevance of air conditioning for 222Radon concentration in shops of the Savona Province, Italy. AB - Radon (222Rn) concentration was evaluated in shops of the Savona Province, Italy, between summer 2002 and winter 2002-2003. The main characteristics of each shops were recorded through a questionnaire investigating the ventilation rate and factors related to 222Rn precursors in the soil and the construction materials. The main variables that were related to radon concentration were the following: age of the building, level of the shop above ground, season of the year, wind exposure, active windows, and type of heating system. Shops equipped with individual air heating/conditioning systems exhibited radon concentrations that were three times higher than those of shops heated by centralized furnaces. Our data indicate that the level of pollution in the shops was of medium level, with an expected low impact on the salespersons' health. Only in wintertime, the action level of 200 Bq m(-3) for the confined environment was reached in 10 shops equipped with individual air heating/conditioning systems. PMID- 15893367 TI - Characterization of endoplasmic reticulum-associated degradation of a protein S mutant identified in a family of quantitative protein S deficiency. AB - INTRODUCTION: Misfolded and unassembled glycoproteins are eliminated from the endoplasmic reticulum (ER) lumen by the ER-associated degradation (ERAD). We previously identified a Tyr595Cys (Y595C) mutation of protein S (PS) in a family of a quantitative PS deficiency. The mutation causes intracellular degradation and decreased secretion of the Y595C mutant PS. The aim of the present study was to further characterize the molecular basis of the intracellular degradation of the mutant. MATERIALS AND METHODS: We stably expressed the mutant in mammalian cells, and analyzed the intracellular localization of the protein. The intracellular degradation pathway was determined by pulse-chase analyses in the presence of various inhibitors of ERAD. RESULTS AND CONCLUSIONS: Endoglycosidase H digestion and immunofluorescence staining revealed the mutant being retained in the ER. Epoxomicin, a potent and specific proteasome inhibitor, and Ala-Ala-Phe CH(2)Cl (AAF), an inhibitor of tripeptidyl peptidase II (TPPII), suppressed the intracellular degradation of the mutant by about 65% and 50%, respectively. When epoxomicin was combined with AAF, the inhibitory effect was substantially enhanced. Although castanospermine, an inhibitor of glucosidases I and II, did not affect the degradation, kifunensine, an inhibitor of ER mannosidase I, suppressed it. Thus, it appears that the Y595C mutant is degraded through more than one pathway of ERAD, including the proteasome-dependent pathway and an alternate proteasome-independent pathway where proteases such as TPPII may be involved. Production of the critical B isoform of Man(8)GlcNAc(2) targets the mutant for ERAD, however, the interaction with calnexin/calreticulin through monoglucosylated oligosaccharides may not be required for the degradation of the mutant. PMID- 15893368 TI - Intimatan (dermatan 4,6-O-disulfate) prevents rethrombosis after successful thrombolysis in the canine model of deep vessel wall injury. AB - INTRODUCTION: Intimatan (dermatan 4,6-O-disulfate), a heparin cofactor II (HCII) agonist, inhibits both the fluid phase and thrombus bound thrombin. The efficacy of Intimatan as an adjunctive anticoagulant during thrombolysis was evaluated in the canine model of arterial injury. MATERIALS AND METHODS: After forming an occlusive thrombus in the right carotid artery (RCA), twenty-one dogs were administered recombinant tissue plasminogen activator (rt-PA) intra-arterially to achieve thrombolysis in the presence of either 0.9% NaCl or Intimatan (9 mg/kg bolus+300 mug/kg/min i.v. infusion). Next, the left carotid arteries (LCA) of the same animals were injured in the presence of either Intimatan or 0.9% NaCl. RESULTS: The incidence of RCA rethrombosis between the Intimatan and control groups was 2/9 and 8/12, respectively. The quality of RCA blood flow, i.e., patency score (Scale of 0-3, i.e., no flow to high flow, respectively), was 2.3+/ 0.4 (Intimatan) versus 0.9+/-0.4 (0.9% NaCl). The incidence of primary thrombosis was determined among the groups as 0/9 (Intimatan) versus 7/12 (0.9% NaCl); the patency score was 2.8+/-0.1 (Intimatan) versus 0.9+/-0.4 (0.9% NaCl). Intimatan resulted in a >90% ex vivo inhibition of gamma-thrombin-induced platelet aggregation whereas 0.9% NaCl had no inhibitory effect. Clot-bound thrombin activity was reduced significantly by Intimatan. Intimatan induced <2-fold change in aPTT and bleeding time (BT) when corrected for the 0.9% NaCl group. CONCLUSIONS: Intimatan significantly reduces the incidence of both primary and secondary arterial thrombosis while maintaining a high-grade vessel patency score with only moderate increases in BT and aPTT. PMID- 15893369 TI - Demographic and genetic characteristics of patients with borderline ovarian tumors as compared to early stage invasive ovarian cancer. AB - OBJECTIVE: Evaluation whether Jewish founder mutations in BRCA predispose to borderline tumors as they do to early invasive ovarian cancers. METHODS: All Jewish women with borderline or invasive ovarian tumors, diagnosed over a 5-year period (1994-1999), were identified in the frame of a nationwide epidemiological study on ovarian cancer in Israel. Out of a total of 1489 patients, 1269 were interviewed; of them 256 (20.2%) patients were identified with stage I and II invasive epithelial ovarian tumors, and 233 (18.3%) patients were identified with borderline tumors. All patients underwent interviews, and blood or tissue samples from 117 borderline tumors and 161 early stage invasive tumors were analyzed for the presence of the 185delAG and 5382insC BRCA1, and the 6174delT BRCA2 Jewish founder mutations. RESULTS: Patients with borderline tumors were younger at diagnosis, and more frequently of the serous type (P < 0.001) as compared to patients with early stage ovarian cancer. Prevalence of Jewish founder mutations in BRCA1 and BRCA2 was only 4.3% of patients with borderline tumors as compared to 24.2% of patients with early stage ovarian cancer (P = 0.001). CONCLUSIONS: This nationwide study comparing patients with early stage borderline and invasive epithelial tumors of the ovary confirms our previous pilot study that showed a lower incidence of BRCA mutations in patients with borderline tumors. Our results suggest that the genetic predisposition and the molecular mechanisms underlying tumor initiation differ between invasive and borderline tumors of the ovary. PMID- 15893370 TI - Dysphoric adolescents' causal explanations and expectancies for approach and avoidance goals. AB - Relatively little research has studied emotional disturbance from a motivational perspective. The current study aimed to investigate personal goal systems at different levels of analysis, namely, personal goals, associated causal explanations and goal expectancies, as a function of dysphoria. Dysphoric (n = 28) and non-dysphoric (n = 28) adolescents (16-18-yr-olds) selected from a larger school sample completed tasks that measured number of approach goals and avoidance goals, causal explanations for goals, and perceived likelihood and control estimates for idiographic goals. As predicted dysphoric adolescents, relative to controls, generated a combination of more avoidance goals and fewer approach goals, and thought of more reasons that explain why goals would not be achieved and fewer reasons that explain why goals would be achieved (irrespective of goal type), and considered avoidance goal outcomes as more likely to occur and approach goal outcomes as less likely to occur. In addition, dysphoric adolescents anticipated less personal control in implementing their goals than controls. Overall, the data identified distinct personal goal systems that function differently in dysphoric and non-dysphoric adolescents. PMID- 15893371 TI - Polyethylene wear particle generation in vivo in an alumina medial pivot total knee prosthesis. AB - Polyethylene wear particle generation is one of the most important factors affecting mid- to long-term results of total knee arthroplasties. It has been reported that the medial pivot total knee prosthesis (MP) design and alumina ceramic femoral component reduce polyethylene wear. The aim of this study is to evaluate in vivo polyethylene wear particle generation in the newly introduced alumina MP, in comparison with a metal MP. Synovial fluid was obtained from 11 knees with alumina MP and 15 knees with metal MP at nine months after the operation. Polyethylene particles were isolated, and examined using scanning electron microscope and image analyzer. Total number of particles in each knee was 7.10+/-2.86x10(6) in alumina (mean+/-standard error), and 5.70+/-2.82x10(7) in metal MP (p=0.048). Particle size (equivalent circle diameter) was 0.78+/-0.04 microm in alumina, and 0.66+/-0.06 microm in metal MP (p=0.120). Particle shape (aspect ratio) was 1.52+/-0.05 in alumina, and 1.88+/-0.11 in metal MP (p=0.014). Apart from the femoral component, the material and manufacturing method of polyethylene insert differed between the two groups, although the sterilization method was the same. Alumina MP generated fewer and rounder polyethylene wear particles than metal MP in early clinical stage, and could potentially reduce prevalence of osteolysis and aseptic loosening. PMID- 15893372 TI - Galanin plasticity in the cholinergic basal forebrain in Alzheimer's disease and transgenic mice. AB - Galanin (GAL) is a biologically active 29 amino acid (30 in humans) which participates in the modulation of several ascending neurotransmitter systems including cholinergic basal forebrain (CBF) neurons, which undergo extensive degeneration in Alzheimer's disease (AD). GAL immunoreactive fibers within the CBF display hypertrophy and hyperinnervate surviving CBF neurons in late AD. Over the years, this unique neuronal plasticity response has been an active area of research for our group. We have examined tissue from a clinically well characterized cohort of retired elderly clergy to determine whether people with mild cognitive impairment display GAL hyperinnervation upon CBF neurons. We found that GAL hyperinnervation is a late stage event and that CBF neuron reduction is not correlated with GAL over expression during prodromal AD. Interestingly, findings from our laboratory using tau immunohistochemistry and single cell gene array technologies suggest that GAL remodeling may influence neurofibrillary tangle formation by altering tau phosphorylation events in CBF neurons in AD. Studies using GAL-tg mice suggest that GAL over expression reduces the cholinergic phenotype but does not produce a frank loss of CBF cells. This phenotypic down regulation of ChAT is reminiscent of the lack of a frank CBF neuron loss in prodromal AD. Moreover, studies using mice transgenic for both the amyloid precursor protein (APP) and presenilin-1 (PS1) bearing AD-related mutations (APPswe/PS1delta9) displayed increased GAL immunoreactive fibers, neurities and plaques in cortex and hippocampus. These fin'dings provide evidence for a mechanistic relationship between amyloidosis and GAL over expression in AD. Understanding GALs role in the clinical and pathological features of AD, may lead to novel drug treatments for this disease. PMID- 15893373 TI - Prognostic value of serum vascular endothelial growth factor (VEGF) and plasma activity of matrix metalloproteinase (MMP) 2 and 9 in lymphoma-affected dogs. AB - Canine non-Hodgkin's lymphoma (NHL) is considered to be a good animal model for its human counterpart; nevertheless, comparative data on neo-angiogenesis are completely lacking. The aim of this study is to investigate the levels of serum vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs) 2 and 9 in lymphoma-affected dogs. Circulating levels of VEGF and both MMP 2 and 9 activities significantly correlate with the WHO sub-stage b prognostic factor; moreover, VEGF at admission have an independent influence on the length of the disease free interval. As in humans, serum VEGF concentration and most likely also MMPs plasma activity have prognostic value in canine NHL spontaneous model. PMID- 15893374 TI - Leukemic cell death induced by antithymocyte globulin. AB - We studied the cytotoxic effects of antithymocyte globulin (ATG) in leukemic cells obtained from five patients with acute T lymphoblastic leukemia or precursor T lymphoblastic leukemia. ATG by itself killed leukemic cells in a dose dependent manner. Simultaneous incubation with human AB or baby rabbit serum resulted in increased cytolysis of leukemic cells. These results suggested the presence of both direct and complement-mediated cytolysis. We also examined apoptotic cell death using Annexin-V. Cell incubation with ATG increased Annexin V binding significantly compared with horse IgG (50.3+/-7.6% versus 95.7+/-1.8%, p = or < 0.0001). However, ATG did not induce apparent DNA fragmentation in a human T-ALL cell line. Neither anti-Fas MoAb (ZB4) nor a broad caspase inhibitor (z-VAD FMK) prevented this increase in Annexin-V binding. These results suggest that ATG induces leukemic cell death in a Fas/Fas-ligand- and caspase-independent manner. PMID- 15893375 TI - Conditioned taste aversion and amygdala lesions in the rat: a critical review. AB - Studies using permanent lesions implicate the amygdala, a recipient of gustatory and viscerosensory information, in taste aversion learning. Reviewing this literature with respect to the location of the lesions and the quality of the behavioral methodology reveals little, if any, involvement of the medial amygdala or central nucleus in conditioned taste aversion. Although a disruption is found following damage to the basolateral region, the attenuated conditioned taste aversion appears to be a consequence of a lesion-induced impairment of neophobia rather than an association formation deficit. The key to understanding the functional significance of the basolateral amygdala in conditioned taste aversion reduces, we believe, to determining the role of this structure in gustatory neophobia. PMID- 15893376 TI - Antidepressant effects assessed using behavior maintained under a differential reinforcement-of-low-rate (DRL) operant schedule. AB - Behavior maintained under a differential-reinforcement-of-low-rate (DRL) 72-s operant schedule, which reinforces responses with interresponse times greater than 72 s, exhibits a rather unique sensitivity to antidepressant drugs. Antidepressants from a number of pharmacological classes, including tricyclic antidepressants, selective serotonin or norepinephrine reuptake inhibitors, monoamine oxidase inhibitors, as well as a number of atypical antidepressants and putative antidepressants, reduce response rate and increase reinforcement rate of rats under this schedule. These effects are observed acutely but persist or are augmented with repeated treatment. By contrast, drugs from a number of other psychotherapeutic classes do not, in general, produce similar effects. This includes anxiolytic, sedative, stimulant, opioid, antihistaminic, and anticholinergic drugs, which can produce false positive results in some preclinical tests for antidepressant efficacy. There are conflicting data regarding the utility of DRL behavior for discriminating the effects of antidepressant and antipsychotic drugs. This results in part from methodological differences among studies, but likely also reflects the overlap between the neuropharmacological and clinical effects of some antipsychotic and antidepressant drugs. DRL behavior also has proven useful for identifying neurochemical and neuroanatomical mediators of antidepressant effects on behavior. Consistent with clinical data, it appears that activation of noradrenergic or serotonergic systems provides for parallel means of producing antidepressant-like effects on DRL behavior. Finally, the results of studies using DRL behavior highlight important roles for central beta-1 adrenergic receptors, as well as 5-HT1A, 5-HT1B, 5-HT2A, and 5-HT2C receptors, in the mediation of antidepressant-like behavioral effects. PMID- 15893377 TI - What should animal models of depression model? AB - In this article, we discuss what animal models of depression should be attempting to 'model'. One must first determine if the goal is to model the regulatory mechanisms by which antidepressant treatments alleviate the various symptoms of depression, or to model the dysregulatory mechanisms underlying the etiology of those symptoms. When modeling the mechanisms of antidepressant effects, a key feature that is often overlooked is the time course required for behavioral efficacy. Even in the clinical literature, there is considerable confusion and inconsistency in defining and identifying 'time of onset' of clinical effect. Although the 'therapeutic lag' may not be as long as has been commonly believed, it does occur. Observable improvement in either global symptomatology or specific symptoms becomes evident after 7-14 days of treatment, and more complete recovery takes considerably longer. Thus, any model addressing potential mechanisms of antidepressant action should exhibit a similar time-dependency. Second, whether attempting to address mechanisms underlying behavioral effects of antidepressants, or the neurobiological substrates underlying the development and manifestation of depression, it is essential to recognize that the syndrome of depression is a diagnostic construct that includes a variety of disparate symptoms, some of which may be related mechanistically, and others that may not be specific to depression, but may cut across categorical diagnostic schemes. Further, it is critical to recognize the close relationship of depression and anxiety. Psychological studies have suggested that the myriad symptoms of depression and anxiety may be subsumed within a more limited number of distinct behavioral dimensions, such as negative affect (neuroticism), positive affect, or physiologic hyperarousal. These dimensions may be related to the functioning of specific neurobiological systems. Thus, rather than trying to recreate or mimic the entire spectrum of symptoms comprising the syndrome of depression, it may be more informative to develop animal models for these behavioral dimensions. Such models may then provide access not only to the neural regulatory mechanisms underlying effective antidepressant treatment, but may also provide clues to the processes underlying the development and manifestation of depression. PMID- 15893378 TI - The programming of individual differences in defensive responses and reproductive strategies in the rat through variations in maternal care. AB - There are profound maternal effects on individual differences in defensive responses and reproductive strategies in species ranging literally from plants to insects to birds. Maternal effects commonly reflect the quality of the environment and are most likely mediated by the quality of the maternal provision (egg, propagule, etc.), which in turn determines growth rates and adult phenotype. In this paper, we review data from the rat that suggest comparable forms of maternal effects on both defensive responses to threat and reproductive behavior and which are mediated by variations in maternal behavior. Ultimately, we will need to contend with the reality that neural development, function and health are defined by social and economic influences. PMID- 15893379 TI - Biochemistry, biology and chemistry of the 5-lipoxygenase product 5-oxo-ETE. AB - 5-Oxo-ETE (5-oxo-6,8,11,14-eicosatetraenoic acid) is an arachidonic acid metabolite formed by the oxidation of 5S-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE) by 5-hydroxyeicosanoid dehydrogenase (5-HEDH), a microsomal enzyme found in leukocytes and platelets. 5-HEDH is highly selective for 5S-HETE, and displays little activity for other monohydroxy metabolites of arachidonic acid. The synthesis of 5-oxo-ETE requires NADP(+) and can be stimulated by activation of the respiratory burst and by oxidative stress. 5-Oxo-ETE is a chemoattractant for eosinophils and neutrophils, and elicits a variety of responses in these cells, including actin polymerization, calcium mobilization, integrin expression, and degranulation. Its primary target appears to be the eosinophil, and among lipid mediators it is the strongest chemoattractant for these cells. It is also a chemoattractant for monocytes and stimulates the proliferation of prostate tumor cells. Its actions are mediated by a G(i) protein-coupled receptor (OXE receptor) that is highly expressed by eosinophils>neutrophils>monocytes. When administered in vivo in both humans and rodents it elicits tissue eosinophilia, suggesting that it may be an important mediator in allergic diseases such as asthma, and that the development of drugs designed to prevent its formation or effects may be useful therapeutic agents in these diseases. PMID- 15893380 TI - Coenzyme A: back in action. AB - Coenzyme A (CoA) is a ubiquitous essential cofactor that plays a central role in the metabolism of carboxylic acids, including short- and long-chain fatty acids. In the last few years, all of the genes encoding the CoA biosynthetic enzymes have been identified and the structures of several proteins in the pathway have been determined. CoA is assembled in five steps from pantothenic acid and pathway intermediates are common to both prokaryotes and eukaryotes. In spite of the identical biochemistry, remarkable sequence differences among some of the prokaryotic and eukaryotic enzymes have been revealed by comparative genomics. Renewed interest in CoA has arisen from the realization that the biosynthetic pathway is a target for antibacterial drug discovery and from the unexpected association of a human neurodegenerative disorder with mutations in pantothenate kinase. The purpose of this review is to integrate previous knowledge with the most recent findings in the genetics, enzymology and regulation of CoA biosynthesis in bacteria, plants and mammals. PMID- 15893381 TI - Effect of intravenous magnesium sulphate in reducing irritability and restlessness in pure and polysubstance opiate detoxification. AB - The aim of this double-blind, placebo-controlled study was to evaluate the efficacy of intravenous magnesium sulphate (MgSO(4)) on the need for chlormethiazole in pure or polysubstance opiate detoxification. Forty-one inpatients suffering from pure and polysubstance opiate dependence were treated with morphine sulphate pentahydrate in a gradual detoxification program. Morphine reduction took about 11 days. Additionally, 5% MgSO(4) was administered intravenously to the intervention group (Mg group, n=22) over 24 h by perfusor (150-200 mg MgSO(4)/h; plasma level of 2.36+/-0.29 mmol/l), whereas NaCl 0.9% was intravenously administered in the placebo group (n=19). In case of withdrawal symptoms (irritability, restlessness, and insomnia), patients received chlormethiazole p.o. Our hypothesis that the need for chlormethiazole would be decreased by adjunctive administration of Mg was not confirmed in our study population (2180 mg/day in the Mg group vs. 2360 mg/day in the placebo group). There was neither a difference in the quantity of chlormethiazole required nor a difference in the severity of withdrawal symptoms measured with the Wang scale between the two comparison groups. We observed that calcium plasma levels decreased and phosphate plasma levels increased significantly during intravenous therapy with Mg. Despite promising pilot studies, the administration of Mg did not enable a dose reduction of tranquilizing medication (chlormethiazole) in pure and polysubstance opiate detoxification. PMID- 15893382 TI - Adolescent anorexia nervosa patients have a discrepancy between neurophysiological responses and self-reported emotional arousal to psychosocial stress. AB - In both clinical practice and research, eating disorder patients are reported to have difficulties in identifying and describing their feelings. They are often described as being unaware of the linkage between their feelings and their behavior. The present study experimentally induced emotions in adolescent anorexia nervosa (AN) patients to examine both self-reported emotional arousal and neurophysiological responses. A group of 10 AN patients and a group of 22 healthy controls (HC) were compared with respect to changes in self-reported emotional arousal and neurophysiological responses, heart rate (HR) and HPA-axis response (cortisol in saliva) during a public speaking test inducing anxious stress. The AN group reported higher levels of anxiety, as a result of stress, but this was not reflected in their HR or cortisol response. By contrast, in the HC group higher levels of self-reported anxiety coincided with clear increases in HR and cortisol. The data indicate that AN patients, in contrast to healthy individuals, show a discordance between self-reported emotional and neurophysiological arousal during psychosocial stress. PMID- 15893383 TI - Are there common mechanisms in sensation seeking and reality distortion in schizophrenia? A study using memory event-related potentials. AB - A growing literature suggests that the characteristics of sensation seeking and reality distortion expressed in schizophrenia share several mechanisms. In a previous study, the comparison of patients with high vs. low reality distortion using event-related potentials (ERPs) recorded in a recognition memory task for unfamiliar faces identified neural and cognitive anomalies specifically related to the expression of these symptoms. As a follow-up, this study investigated the ERP correlates of sensation seeking in schizophrenia using the same recognition memory protocol. ERPs have been recorded in controls (N=21) and schizophrenia patients separated into high (HSS; N=13) and low (LSS; N=17) scorers according to Zuckerman's Sensation Seeking Scale. The results show a reduced P2a that was found unrelated to reality distortion in the previous study of reality distortion. It identifies interference inhibition impairment as being specifically related to sensation seeking. On the other hand, HSS scorers display enhanced fronto-central and normal P600 effects also found in high reality distortion patients. These results indicate inappropriate context processing and mnemonic binding common to sensation seeking and reality distortion. LSS scorers also display a reduced temporal N300 similar to that found in low reality distortion patients. This anomaly could reflect the lower reactivity to emotionally significant stimuli that underlies anhedonia symptoms. Finally, the N400 effect and a late frontal effect are found in both HSS and LSS. Since they were unrelated to reality distortion, these indices have been related to basic aspects of schizophrenia, e.g., deficient knowledge integration, or other mechanisms, e.g. anxiety or impulsivity. In summary, the present study examines the strategy of investigating variables, such as temperamental characteristics, in addition to symptoms, to show how discrete impairments may contribute to the expression of the illness. PMID- 15893384 TI - Poor antisaccade performance in schizophrenia: an inhibition deficit? AB - The antisaccade task appears to be particularly suitable for analyzing processes involved in executive control of action. Schizophrenic patients show enhanced rates of erroneous reflexive saccades in this task. This is commonly interpreted as a failure of inhibitory mechanisms. The role of volitional saccade generation is largely neglected in these accounts. In this study, experimental variations of the antisaccade task were applied to manipulate the contribution of volitional processes on antisaccade performance. Fifteen patients with a diagnosis of schizophrenia and 15 healthy control participants performed antisaccade tasks requiring them to look to the mirror location of a peripheral visual stimulus at the onset of this stimulus (standard antisaccade task) or after a brief delay (delayed antisaccade task). As expected, schizophrenic patients showed more reflexive saccade errors than controls. In the delay conditions, reflexive errors decreased, and this effect was significantly stronger in schizophrenic patients. Latencies of correct antisaccades tended to be longer in patients than in control participants. The results suggest that the generation of voluntary saccades is at least in part responsible for the antisaccade deficit in schizophrenic patients. More comprehensive models to account for executive deficits in the antisaccade task must be considered. PMID- 15893385 TI - Abrogation of DTH response and mitogenic lectin- and alloantigen-induced activation of lymphocytes by calcium inhibitors TMB-8 and BAPTA-AM. AB - In vitro treatment of mouse lymphocytes with an intracellular calcium chelator BAPTA-AM significantly decreased lectin Concanavalin-A (Con-A)-induced and mixed lymphocyte reaction (MLR) mediated, alloantigen-induced lymphocyte activation as indicated by decreased percentage of lymphoblasts among the BAPTA-treated lymphocytes. In vivo treatment of mice with intracellular Ca(2+) antagonist TMB-8 was found to substantially impair delayed type hypersensitivity (DTH; cell mediated immune) response, as indicated by decreased footpad swelling on tuberculin challenge of mice sensitized with BCG, after a single treatment with a low dose of 0.01mg of TMB-8 per mouse. Interestingly, a second injection of a higher dose of TMB-8 (0.1mg per mouse) resulted in very significant (p=0.001) abrogation of DTH as indicated by complete absence of swelling of foot pad after PPD challenge in BCG-primed treated mice. All mice in this group showed fully impaired DTH response. Lymphocytes of allosensitized mice gave a significantly higher (p<0.05) MLR response than the naive mice. However, a single treatment of allosensitized mice with 0.1mg TMB-8 resulted into a lower MLR response, comparable in magnitude to that of untreated naive mice. PMID- 15893386 TI - Succinylcholine-induced masseter muscle rigidity during bronchoscopic removal of a tracheal foreign body. AB - Masseter muscle rigidity during general anesthesia is considered an early warning sign of a possible episode of malignant hyperthermia. The decision whether to continue or discontinue the procedure depends on the urgency of the surgery and severity of masseter muscle rigidity. Here, we describe a case of severe masseter muscle rigidity (jaw of steel) after succinylcholine (Sch) administration during general anesthetic management for rigid bronchoscopic removal of a tracheal foreign body. Anesthesia was continued uneventfully with propofol infusion while all facilities were available to detect and treat malignant hyperthermia. PMID- 15893387 TI - Rapid and reproducible infectivity end-point titration of virulent phage in a microplate system. AB - The standard method for measuring the number of infectious phages in solution has traditionally been the plaque forming assay. An alternative method is described where the number of lytic, infectious phages is determined in an endpoint titration assay adapted for a microplate system. In this model system, susceptible Escherichia coli B6 at a density of 4 x 10(7) cells/ml, were mixed with an equal volume (100 microl) of PhiX174 diluted serially in a microtest plate. After 3h of incubation on a microplate shaker the endpoint was determined spectrophotometrically and calculated according to the method of Reed and Muench. A well was considered positive for infection if the OD630-value was < or = 10% compared to the OD630-value of the negative control of uninfected cells. ID50 titers were 2.5x higher than the PFU-titers (CV 15%) and the intra assay reproducibility revealed a CV of 9%. The method has several advantages as compared with the conventional PFU-titration. It is less time and material consuming with the possibility to assess several samples at the same time. PMID- 15893388 TI - The role of reactive oxygen species in TNFalpha-dependent expression of the receptor for advanced glycation end products in human umbilical vein endothelial cells. AB - Engagement of the receptor for advanced glycation end products (RAGE) by its signal transduction ligands is implicated in the development and progression of atherosclerosis. TNFalpha, a proinflammatory cytokine, is a potent inducer of RAGE expression in endothelial cells. In the present study, we demonstrate that reactive oxygen species (ROS) generated by TNFalpha stimulated human umbilical vein endothelial cells (HUVECs) induce RAGE expression. The complex III of mitochondrial respiratory chain appears to be the primary source of ROS. The gp91phox subunit of NADPH oxidase appears to be the source of ROS that induces TNFalpha-dependent mitochondrial ROS generation and subsequent RAGE expression. We also demonstrate that the ROS-mediated RAGE induction occurs via activation of NF-kappaB, a proinflammatory transcription factor. Thus, stimulation of HUVECs by TNFalpha evokes the following sequence of events: stimulation of NADPH oxidase - > generation of ROS --> activation of the mitochondrial respiratory chain --> stimulation of NF-kappaB activity --> induction of RAGE expression. PMID- 15893389 TI - SNAREs and traffic. AB - SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) are now generally accepted to be the major players in the final stage of the docking and the subsequent fusion of diverse vesicle-mediated transport events. The SNARE-mediated process is conserved evolutionally from yeast to human, as well as mechanistically and structurally across different transport events in eukaryotic cells. In the post-genomic era, a fairly complete list of "all" SNAREs in several organisms (including human) can now be made. This review aims to summarize the key properties and the mechanism of action of SNAREs in mammalian cells. PMID- 15893390 TI - Fluorescence in situ hybridisation coupled to ultra small immunogold detection to identify prokaryotic cells using transmission and scanning electron microscopy. AB - We describe a method based on fluorescence in situ hybridisation (FISH) that allows the identification of individual cells by electron microscopy. We hybridised universal and specific fluorescein-labelled oligonucleotide probes to the ribosomal RNA of prokaryotic microorganisms in heterogeneous cell mixtures. We then used antibodies against fluorescein coupled to sub-nanometer gold particles to label the hybridised probes in the ribosome. After increasing the diameter of the metal particles by silver enhancement, the specific gold-silver signal was visualised by optical microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). It is the first time that SEM is applied to the detection of gold nanoparticles hybridised to an intracellular target, such as the ribosome. The possibility to couple phylogenetic identification by FISH to cell surface and ultrastructure observation at electron microscopy resolution has promising potential applications in microbial ecology. PMID- 15893391 TI - Multicenter evaluation of the nitrate reductase assay for drug resistance detection of Mycobacterium tuberculosis. AB - The performance of the nitrate reductase assay was evaluated in a multicenter laboratory study to detect resistance of Mycobacterium tuberculosis to the first line anti-tuberculosis drugs rifampicin, isoniazid, ethambutol and streptomycin using a set of coded isolates. Compared with the gold standard proportion method on Lowenstein-Jensen medium, the assay was highly accurate in detecting resistance to rifampicin, isoniazid and ethambutol with an accuracy of 98%, 96.6% and 97.9%, respectively. For streptomycin, discrepant results were obtained with an overall accuracy of 85.3%. The assay proved easy to be implemented in countries with limited laboratory facilities. PMID- 15893392 TI - Comparison of drug resistance genotypes between Beijing and non-Beijing family strains of Mycobacterium tuberculosis in Korea. AB - The frequency of resistance genotypes among Beijing and non-Beijing strains was compared using a reverse blot hybridization assay to detect mutations within genes associated with rifampicin (rpoB) and isoniazid (katG, inhA, and ahpC) resistance. Of the 743 Mycobacterium tuberculosis isolates, 569 (77%) belonged to Beijing family. The proportion of Beijing strains was significantly higher among MDR-TB isolates than among drug-susceptible strains (82% vs. 72%, p<0.01). Genotype analysis of the rpoB gene revealed significantly lower rates of the Ser531Leu mutation rate among Beijing vs. non-Beijing MDR-TB strains (41% vs. 66%, p<0.005). While the mutation for Ser315Thr in the katG gene was more common among Beijing vs. non-Beijing family strains (65% vs. 50%, p<0.01), the mutation rate of promoter region of the inhA gene was lower among Beijing strains compared with non-Beijing strains (14% vs. 25%, p<0.05). Reverse hybridization successfully detected over 80% of isoniazid-resistant strains and over 92% of rifampicin-resistant strains among Korean isolates. Significant differences in mutation rates in the rpoB, katG, and inhA genes between Beijing strains and non Beijing strains could explain discrepancies in mutation rates of genotypes in different countries. Reverse hybridization was useful for rapid detection of isoniazid and rifampicin resistant strains. PMID- 15893393 TI - Methods for the recovery, isolation and detection of Cryptosporidium oocysts in wastewaters. AB - This correspondence describes the successful development of methods for the recovery, isolation and detection of Cryptosporidium oocysts in wastewater and biosolids. Wastewater from one plant was used to optimize methods in raw influent as well as primary, secondary and tertiary effluents. Raw influents and primary effluents were concentrated using centrifugation followed by isolation of Cryptosporidium oocysts using immunomagnetic separation (IMS) and detection of recovered organisms using epifluorescence microscopy. Mean oocyst recovery in raw influent was 29.2+/-12.8% and 38.8+/-27.9% in primary effluent at three sample volumes tested. Secondary and tertiary effluents were analyzed using a modified Method 1622 resulting in mean oocyst recoveries of 53.0+/-19.2% and 67.8+/-4.4%, respectively. In biosolids with approximately 10% total solids, mean oocyst recovery was 43.9+/-10.1% using IMS with a 5 g (wet weight) sample size. Due to the variability in these matrices, an internal microbiological standard was incorporated to serve as a tool for method performance. PMID- 15893394 TI - Landscape phage probes for Salmonella typhimurium. AB - We selected from landscape phage library probes that bind preferentially Salmonella typhimurium cells compared with other Enterobacteriaceae. The specificity of the phage probes for S. typhimurium was analyzed by the phage capture test, the enzyme-linked immunosorbent assay (ELISA), and the precipitation test. Interaction of representative probes with S. typhimurium was characterized by fluorescence-activated cell sorting (FACS), and fluorescent, optical and electron microscopy. The results show that the landscape phage library is a rich source of specific and robust probes for S. typhimurium suitable for long-term use in continuous monitoring devices and biosorbents. PMID- 15893395 TI - Rapid identification of dairy lactic acid bacteria by M13-generated, RAPD-PCR fingerprint databases. AB - About a thousand lactic acid bacteria (LAB) isolated from dairy products, especially cheeses, were identified and typed by species-specific PCR and RAPD PCR, respectively. RAPD-PCR profiles, which were obtained by using the M13 sequence as a primer, allowed us to implement a large database of different fingerprints, which were analysed by BioNumerics software. Cluster analysis of the combined RAPD-PCR fingerprinting profiles enabled us to implement a library, which is a collection of library units, which in turn is a selection of representative database entries. A library unit, in this case, can be considered to be a definable taxon. The strains belonged to 11 main RAPD-PCR fingerprinting library units identified as Lactobacillus casei/paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus helveticus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus brevis, Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus and Lactococcus lactis. The possibility to routinely identify newly typed, bacterial isolates by consulting the library of the software was valued. The proposed method could be suggested to refine previous strain identifications, eliminate redundancy and dispose of a technologically useful LAB strain collection. The same approach could also be applied to identify LAB strains isolated from other food ecosystems. PMID- 15893396 TI - Low prevalence of antibodies against heat shock protein 10 of Chlamydophila pneumoniae in patients with coronary heart disease. AB - In this study the prevalence of antibodies against the heat shock protein 10 (HSP10) of Chamydophila pneumoniae (CP) (as assessed by ELISA) in patients with coronary heart disease (CHD) and seropositive or seronegative to CP, as assessed by microimmunofluorescence (MIF), was investigated. The controls were age- and sex-matched healthy subjects. The HSP10 preparation used throughout this study was a 6-his-tagged recombinant protein preliminarily shown to be immunogenic in mice. Low level IgG reactivity against CP-HSP10 was detected in 19 out of 200 and 5 out of 100 CHD patients and controls, respectively. No IgM or IgA isotypes were found. Furthermore, there was no difference in the frequency or level of anti HSP10 IgG between CP-positive and CP-negative sera either in patients (11/140=7.9% vs. 8/60=13%) or in healthy subjects (3/40=7.5% vs. 2/60=3.3%). Overall, our data indicate that CP-HSP10, at variance with CP-HSP60, to which it is genetically and physiologically linked, should not be regarded as a major expressed immunogen or a marker of infection by CP in CHD patients. PMID- 15893397 TI - Validation of the thoracic impedance derived respiratory signal using multilevel analysis. AB - The purpose of the current study was to validate the change in thoracic impedance (dZ) derived respiratory signal obtained from four spot electrodes against incidental spirometry. Additionally, a similar validation was performed for a dual respiratory belts signal to compare the relative merit of both methods. Participants were 38 healthy adult subjects (half male, half female). Cross method comparisons were performed at three (paced) respiration frequencies in sitting, supine and standing postures. Multilevel regression was used to examine the within- and between-subjects structure of the relationship between spirometric volume and the respiratory amplitude signals obtained from either dZ or respiratory belts. Both dZ derived respiratory rate and dual belts derived respiratory rate accurately reflected the pacing frequencies. For both methods, fixed factors indicated acceptable but posture-specific regression on spirometric volume. However, random factors indicated large individual differences, which was supported by variability of gain analyses. It was concluded that both the dZ and dual belts methods can be used for measurement of respiratory rate and within subjects, posture-specific, changes in respiratory volume. The need for frequent subject-specific and posture-specific calibration combined with relatively large measurement errors may strongly limit the usefulness of both methods to assess absolute tidal volume and minute ventilation in ambulatory designs. PMID- 15893398 TI - Long-term depression in rat CA1-subicular synapses depends on the G-protein coupled mACh receptors. AB - The subiculum, which is the primary target of CA1 pyramidal neurons and sending efferent fibres to many brain regions, serves as a hippocampal interface in the neural information processes between hippocampal formation and neocortex. Long term depression (LTD) is extensively studied in the hippocampus, but not at the CA1-subicular synaptic transmission. Using whole-cell EPSC recordings in the brain slices of young rats, we demonstrated that the pairing protocols of low frequency stimulation (LFS) at 3 Hz and postsynaptic depolarization of -50 mV elicited a reliable LTD in the subiculum. The LTD did not cause the changes of the paired-pulse ratio of EPSC. Furthermore, it did not depend on either NMDA receptors or voltage-gated calcium channels (VGCCs). Bath application of the G protein coupled muscarinic acetylcholine receptors (mAChRs) antagonists, atropine or scopolamine, blocked the LTD, suggesting that mAChRs are involved in the LTD. It was also completely blocked by either the Ca2+ chelator BAPTA or the G-protein inhibitor GDP-beta-S in the intracellular solution. This type of LTD in the subiculum may play a particular role in the neural information processing between the hippocampus and neocortex. PMID- 15893399 TI - GInaFiT, a freeware tool to assess non-log-linear microbial survivor curves. AB - This contribution focuses on the presentation of GInaFiT (Geeraerd and Van Impe Inactivation Model Fitting Tool), a freeware Add-in for Microsoft Excel aiming at bridging the gap between people developing predictive modelling approaches and end-users in the food industry not familiar with or not disposing over advanced non-linear regression analysis tools. More precisely, the tool is useful for testing nine different types of microbial survival models on user-specific experimental data relating the evolution of the microbial population with time. As such, the authors believe to cover all known survivor curve shapes for vegetative bacterial cells. The nine model types are: (i) classical log-linear curves, (ii) curves displaying a so-called shoulder before a log-linear decrease is apparent, (iii) curves displaying a so-called tail after a log-linear decrease, (iv) survival curves displaying both shoulder and tailing behaviour, (v) concave curves, (vi) convex curves, (vii) convex/concave curves followed by tailing, (viii) biphasic inactivation kinetics, and (ix) biphasic inactivation kinetics preceded by a shoulder. Next to the obtained parameter values, the following statistical measures are automatically reported: standard errors of the parameter values, the Sum of Squared Errors, the Mean Sum of Squared Errors and its Root, the R(2) and the adjusted R(2). The tool can help the end-user to communicate the performance of food preservation processes in terms of the number of log cycles of reduction rather than the classical D-value and is downloadable via the KULeuven/BioTeC-homepage at the topic "Downloads" (Version 1.4, Release date April 2005). PMID- 15893400 TI - New developments in tracing neural circuits with herpesviruses. AB - Certain neurotropic viruses can invade the nervous system of their hosts and spread in chains of synaptically connected neurons. Consequently, it is possible to identify entire hierarchically connected circuits within an animal. In this review, we discuss the use of neurotropic herpesviruses as neuronal tract tracers. Although a variety of tract tracing viruses are available, each with its own unique infection characteristics, we focus on the widespread use of attenuated strains of pseudorabies virus (PRV), a swine herpesvirus with a broad host range. In particular, we focus on new applications of PRV for tract tracing including use of multiple infections by PRV reporter viruses to test for circuit convergence/divergence within the same animal. We provide examples of these combined application techniques within the context of an animal model to study the naturally occurring reversal of seasonal obesity in Siberian hamsters. PMID- 15893401 TI - Infection and activation of bursal macrophages by virulent infectious bursal disease virus. AB - In this study the effect of infectious bursal disease virus (IBDV) on bursal macrophages during the acute phase of the infection was examined. Specific pathogen-free (SPF) chickens were exposed to virulent IBDV and bursal adherent cells were examined by immunohistochemisrty and RT-PCR for virus infection and by real-time quantitative RT-PCR (qRT-PCR) for mRNA transcripts of proinflammatory cytokines and iNOS. Viral genome was detected in bursal macrophages at 3, 5 and 7 days post-infection (dpi). Immuno-histochemical staining revealed double positive cells for KUL01 (macrophage marker) and intracellular viral proteins, showing viral replication in bursal macrophages of infected chickens. We noted a significant decrease in the total number of bursal macrophages in infected chickens, probably due to the lysis of infected cells. However, likely due to extensive necrosis of B cells, the relative proportion of bursal macrophages was significantly higher (P<0.05) in infected birds at 3 and 5 dpi than in controls. Among the cytokines examined, IL-6 showed the greatest upregulation (100-fold increase) at 3 dpi. Expression of IL-1beta was maximum at 3 dpi whereas IL-18 expression was highest at 1 dpi. Enhanced expression of iNOS mRNA was observed at 5 dpi. Increased expression of the proinflammatory cytokines and iNOS correlated well with the presence of the inflammatory response in the infected bursa. These data suggested that B cells may not be the sole targets for the virus; macrophages and possibly other cells may serve as host for IBDV. PMID- 15893402 TI - Intrahepatic CD8+ T-lymphocyte response is important for therapy-induced viral clearance in chronic hepatitis B infection. AB - BACKGROUND/AIMS: To determine which immune cells contribute to HBV-clearance during antiviral therapy, we performed a longitudinal analysis of intrahepatic immune cells during interferon-alpha therapy of chronic HBV-patients using the FNAB technique. METHODS: Twenty chronic HBeAg+-patients were treated with pegylated alpha-interferon combined with lamivudine or placebo for 52 weeks. FNAB and blood specimens were obtained at week 0, 2, 8 and 52. CD4+- and CD8+ T lymphocytes, CD56+ cells, IFNgamma and granzyme B (GrB) were immunocytochemically quantified. RESULTS: The relative numbers of CD56+ cells and CD8+ T-lymphocytes were significantly higher in FNAB compared to blood at all time-points. Responders (n=9) exhibited significant increases in intrahepatic CD8+ and CD8+GrB+ lymphocytes, a small elevation in CD8+IFNgamma+ T-lymphocytes, no change in CD4+ T-lymphocytes, and a decrease in intrahepatic CD56+ cells during the first weeks of therapy. In non-responders (n=11) no significant changes in CD4+- and CD8+ T-lymphocytes and an increase in intrahepatic and CD56+ cells were observed during therapy. CONCLUSIONS: The intrahepatic CD8+ T-lymphocyte, but not the CD4+ T-lymphocyte or NK/NKT-cell response, is important for HBV clearance during interferon-alpha therapy, and the antiviral effect may be mediated by both cytolytic and non-cytolytic mechanisms. PMID- 15893403 TI - Mesothelioma, perspective from the industry. PMID- 15893404 TI - Effect of a selective OX1R antagonist on food intake and body weight in two strains of rats that differ in susceptibility to dietary-induced obesity. AB - An orexin-1 receptor antagonist decreases food intake whereas orexin-A selectively induces hyperphagia to a high-fat diet. In the present study, we evaluated the effect of an orexin antagonist in two strains of rats that differ in their sensitivity to becoming obese while eating a high-fat diet. Male Osborne Mendel (OM) and S5B/Pl (S5B) rats were treated acutely with an orexin-1 receptor antagonist (SB-334867), after adaptation to either a high-fat (56% fat energy) diet or a low-fat (10% fat energy) diet that were equicaloric for protein (24% energy). Ad libitum fed rats were injected intraperitoneally with SB-334867 at doses of 3, 10 or 30 mg/kg, or vehicle at the beginning of the dark cycle, and food intake and body weight were measured. Hypothalamic prepro-orexin and orexin 1 receptor mRNA expression were analyzed in OM and S5B rats fed at a high-fat or low-fat diet for two weeks. SB-334867 significantly decreased food intake in both strains of rats eating the high-fat diet but only in the OM rats eating the low fat diet. The effect was greatest at 12 and 24 h. Body weight was also reduced in OM rats 1d after injection of SB-334867 but not in the S5B rats. Prepro-orexin and orexin-1 receptor expression levels did not differ between strains or diets. These experiments demonstrate that an orexin antagonist (SB-334867) reduces food intake and has a greater effect in a rat strain that is susceptible to dietary induced obesity, than in a resistant strain. PMID- 15893405 TI - Isolation and characterization of antagonist and agonist peptides to the human melanocortin 1 receptor. AB - We identified a large number of peptide mimotopes of the adrenocorticotropic hormone (ACTH) and the alpha-melanocyte stimulating hormone (alpha-MSH) to analyze better the structure-function relationships of these hormones with the human MC1 receptor (hMC1R). We have investigated the use of phage-display technology to isolate specific peptides of this receptor by using three monoclonal anti-ACTH antibodies (mAbs). A library of 10(8) phage-peptides displaying randomized decapeptides was constructed and used to select phage peptides that bind to mAbs. Forty-five phage-peptides have been isolated and from their amino acid sequences, we have identified two consensus sequences, EXFRWGKPA and WGXPVGKP, corresponding to the regions 5-13 and 9-16 of ACTH, respectively. A biological assay on cells expressing the hMC1-R was developed to determine the capacity of phage-peptides to stimulate the receptor. Only two phage-peptides showed detectable activity. Thirty-one peptides were synthesized to analyze their biological effect. We identified two weak agonists, EC50=16 and 11 microM, two strong agonists, EC50=25 and 14 nM and a partial antagonist, IC50=36 microM. This work confirmed the modulator agonist role of the regions 11-12 of alpha-MSH and ACTH, and the importance of the methionine residue at position 4 for the stimulation of the hMC1-R. We also identified analogues of the regions 8-17 of ACTH that exhibited a weak activator effect, and of one analogue of the N terminal regions 1-9 of ACTH and alpha-MSH having a partial antagonist effect. These results may be useful in the development of potential agonists or antagonists of the hMC1R. PMID- 15893406 TI - CSF hypocretin-1/orexin-A concentrations in patients with subarachnoid hemorrhage (SAH). AB - The aim of this study was to examine the role of the hypothalamic hypocretin/orexin system in complications of delayed ischemic neuronal deficit (DIND) resulting from symptomatic vasospasm in patients with aneurysmal subarachnoid hemorrhage (SAH). CSF hypocretin-1/orexin-A levels were measured in 15 SAH patients. DIND complications occurred in seven patients with symptomatic vasospasm. Hypocretin-1/orexin-A levels were low in SAH patients during the 10 days following the SAH event. CSF hypocretin-1/orexin-A levels were lower in patients with DIND complications than in those who did not develop DIND. A significant transient decline in CSF hypocretin-1/orexin-A levels was also observed at the onset of DIND in all patients with symptomatic vasospasm. The reduced hypocretin/orexin production observed in SAH patients may reflect reduced brain function due to the decrease in cerebral blood flow. These results, taken together with recent experimental findings in rats that indicate hypocretin receptor 1 (orexin 1 receptor) mRNA and protein are elevated following middle cerebral artery occlusion, suggest that a reduction in hypocretin/orexin production in SAH and DIND patients is associated with alterations in brain hypocretin/orexin signaling in response to ischemia. PMID- 15893407 TI - Synergistic inhibition of respiration in brain mitochondria by nitric oxide and dihydroxyphenylacetic acid (DOPAC). Implications for Parkinson's disease. AB - The inhibition of mitochondrial respiration by nitric oxide (.NO) at cytochrome c oxidase level has been established as a physiological regulatory mechanism of mitochondrial function. Given, on the one hand, the potential involvement of .NO and dopamine metabolism in mitochondrial dysfunction associated with neurodegeneration and, on the other hand, the reported interaction of .NO with dihydroxyphenylacetic acid (DOPAC), a major mitochondrial-associated dopamine metabolite, we examined the combined effects of .NO and DOPAC on the respiratory chain of isolated rat brain mitochondria. Whereas dopamine or DOPAC induced no measurable effects on the mitochondrial respiration rate, a mixture of .NO with DOPAC inhibited the rate in a way stronger than that exerted by .NO. This effect was noticed with actively respiring (state 3) and resting (state 4) mitochondria. At variance with DOPAC, dopamine failed to potentiate .NO inhibitory effects. The inhibition was dependent on the concentration of both compounds, .NO and DOPAC, and exhibited characteristics similar to those exerted by .NO, namely: it was reversible and dependent on the concentration of oxygen. Analysis of respiratory enzymatic activities demonstrated a selective inhibition at the level of cytochrome c oxidase (complex IV). Insights into the chemical mechanisms underlying the inhibitory effect were inferred from experiments using metmyoglobin (a ligand for .NO and derived species, such as nitroxyl anion) and ferrocyanide (a reductant of .NO, producing nitroxyl anion). Whereas metmyoglobin decreased the inhibition, ferrocyanide potentiated the inhibition. Moreover, a mixture of ferrocyanide with .NO reproduced the effects exerted by the mixture of .NO with DOPAC. The results are consistent with the notion of a reaction of .NO with DOPAC producing a nitric oxide-derived compound(s), which inhibit O2 uptake at the cytochrome oxidase level. Although the mechanism in question remains to be clearly elucidated it is suggested that the .NO/DOPAC-dependent inhibition of cytochrome oxidase may involve nitroxyl anion. The significance of these observations for mitochondrial dysfunction inherent in Parkinson's disease is discussed. PMID- 15893408 TI - MRL/lpr mice have alterations in brain metabolism as shown with [1H-13C] NMR spectroscopy. AB - Cerebral glucose metabolism and cerebral blood flow are altered in patients with lupus who have neuropsychiatric manifestations. However, the dynamics of changes in glucose metabolism remain unclear. The present study was undertaken using 1H and 13C nuclear magnetic resonance (NMR) spectroscopy to determine the rates of incorporation of glucose into amino acids and lactate via cell-specific pathways in mice with lupus. In the well-established MRL/lpr lupus mouse model, 24-week old mice had a significant increase of 30-80% (P<0.001) in total brain glutamine, glutamate and lactate concentrations, while alanine, aspartate, N-acetyl aspartate (NAA) and gamma-aminobutyric acid (GABA) remained unchanged as compared to the congenic MRL+/+control mice. Although succinate concentration was increased in lupus brain, it did not reach statistical significance. Furthermore, 13C isotopomer analysis showed a selective increase of de novo synthesis of lactate from [1-(13)C] glucose through glycolysis resulting in 1.5-fold increased fractional 13C enrichments in lactate in MRL/lpr mice. [4-(13)C] Glutamate, which is synthesized mainly by the neuronal pyruvate dehydogenase, was selectively increased, while [2-(13)C] and [3-(13)C] GABA synthesis were decreased by 25% compared to controls. In accordance with the total concentrations, aspartate synthesis remained unaltered in brains of lupus mice, while alanine synthesis was elevated, indicating increased utilization of alanine. Creatine was unchanged in MRL/lpr mice as compared to controls. An interesting finding was a significant increase (158%, P<0.005) in choline concentration in MRL/lpr mice while the myo inositol concentration remained the same in both groups. Furthermore a significant increase in total brain water content was observed, indicative of possible edema. In conclusion, the cumulative effect of increased brain lactate synthesis, altered glucose metabolism and intracellular glutamine accumulation could be an important mechanism causing brain pathology in SLE. The alteration in metabolites could alter downstream pathways and cause neurological dysfunction. Future NMR spectroscopic studies using stable isotopes and real-time measurements of metabolic rates, along with levels of metabolites in plasma and cerebrospinal fluid, could be valuable in the elucidation of the cerebral metabolic consequences of systemic lupus erythematosis (SLE) in humans. PMID- 15893409 TI - Anaemia increases the risk of dementia in cognitively intact elderly. AB - Although cross-sectional studies found an association between anaemia and dementia, longitudinal studies provided contradictory results. We hypothesize that anaemia might increase the risk of developing dementia because of chronic brain hypo-oxygenation. Using baseline data from a community-based longitudinal study, the Kungsholmen Project, Stockholm, Sweden, we clinically followed 1435 non demented subjects aged 75-95 years for 3 years to detect incident dementia cases (DSM-III-R criteria). Subjects that fulfilled WHO criteria for anaemia, baseline haemoglobin concentration; 130 g/L (men) and 120 g/L (women), had a higher hazard ratios (HR) of developing dementia 3 years later (HR 1.6, 95% CI: 1.1-2.4). In persons with good baseline cognition (MMSE>or=26, n=1139), the association was stronger and still significant after adjustments for conditions potentially related to anaemia and dementia, such as chronic diseases, inflammatory markers, and indicators of nutritional status. The HR was increased even when different haemoglobin cut offs for anaemia definition were used. Thus, anaemia is suggested to be a new potential modifiable risk factor for dementia. PMID- 15893410 TI - Peripheral infection and aging interact to impair hippocampal memory consolidation. AB - We report that a peripheral injection of Escherichia coli produces both anterograde and retrograde amnesia in 24 month old, but not 3 month old rats for memories that depend on the hippocampus, that is, memory of context, contextual fear, and place learning. The anterograde effect was restricted to measures of long-term memory. Short-term memory was not affected, nor did E. coli produce amnesia for auditory-cue fear conditioning. There were no age related effects on memory in vehicle-treated rats. In addition to these age-related cognitive effects of E.coli, we report that it produced a marked increased in IL-1beta levels in the hippocampus, but not in parietal cortex or serum. These findings support the hypothesis that age is a vulnerability factor that increases the likelihood that an immune challenge will produce a cognitive impairment. It is possible that this cognitive vulnerability is mediated by age-related changes in the glial environment that results in an exaggerated brain pro-inflammatory response to infection. PMID- 15893411 TI - [Rejuvenation of the lower third of the face and the neck]. AB - One of the marks of the youth of a face is the absence of ptosis of the cheek and the perfect delimitation of the neck with an angle well defined. The objective of this work is to present our surgical technique for the treatment of the lower third of the face and the neck. After a degreasing of the neck and lower part of the cheek, we carry out an extensive cutaneous separation of these areas. The cervical face lift is performed in different layers. A suspension with posterior direction of the platysma is systematic. It is associated or not, according to the importance with the platysma bands, with an anterior approach or with counter side incisions. A reinforcement of the angles is obtained by a fat grafting. One fills the mandibular ligament and one underlines the relief of the chin. PMID- 15893412 TI - Interaction of anticancer drug mitoxantrone with DNA analyzed by electrochemical and spectroscopic methods. AB - Cyclic voltammetry coupled with different spectroscopic (UV/Vis, fluorescence and Raman) techniques were used to study the interaction of mitoxantrone (MTX), an antitumor drug, with calf thymus DNA in acetate buffer solutions (pH 4.5). The interaction of MTX with DNA could result a considerable decrease in the MTX peak currents and a hypochromic and bathochromic shift in the maximum adsorption bands of MTX as well as the emission quenching in the MTX fluorescence spectra. The variations in the electrochemical and spectral characteristics of MTX indicated MTX bind to DNA by an intercalative mode. This conclusion was reinforced by Raman data. The merely particular vibrations were affected in Raman, suggesting that only a portion of the chromophore of MTX was involved in the intercalation into DNA duplex. These studies are valuable for a better understanding the detailed mode of MTX-DNA interaction, which should be important in deeper insight into the therapeutic efficacy of MTX and design of new DNA targeted drug. PMID- 15893413 TI - Direct observation of resonance tryptophan-to-chromophore energy transfer in visible fluorescent proteins. AB - Visible fluorescent proteins from Aequorea victoria contain next to the fluorophoric group a single tryptophan residue. Both molecules form a single donor-acceptor pair for resonance energy transfer (RET) within the protein. Time resolved fluorescence experiments using tryptophan excitation have shown that RET is manifested by a distinct growing in of acceptor fluorescence at a rate characteristic for this process. In addition, time-resolved fluorescence anisotropy measurements under the same excitation-emission conditions showed a correlation time that is similar to the time constant of the same RET process with the additional benefit of gaining information on the relative orientation of the corresponding transition dipoles. PMID- 15893414 TI - Vessel dilator: most potent of the atrial natriuretic peptides in decreasing the number and DNA synthesis of human squamous lung cancer cells. AB - Within 24 h four peptide hormones, i.e. vessel dilator, long acting natriuretic peptide, kaliuretic peptide, and atrial natriuretic peptide and their intracellular mediator cyclic GMP decreased the number of human squamous lung cancer cells 51, 22, 25, 21, and 30%, respectively. There was not any proliferation in the 3 days following this decrease in cell number. Vessel dilator decreased DNA synthesis 85% in the squamous lung cancer cells. Thus, vessel dilator significantly decreased the number of human squamous lung cancer cells and their DNA synthesis, mediated in part by cyclic GMP, more than other peptide hormones. PMID- 15893415 TI - Evidence for CEA release from human colon cancer cells by an endogenous GPI-PLD enzyme. AB - Elevated carcinoembryonic antigen (CEA) blood levels are found in a wide variety of epithelial neoplasms. The precise mechanism of the spontaneous CEA release from normal and cancer cells has not been established yet. In this study we investigated 'in vitro' the role of an endogenous glycosylphosphatidyl inositol phospholipase D (GPI-PLD) in spontaneous CEA release from human colon carcinoma cells. We detected GPI-PLD-specific transcript expression in four human colorectal tumor cell lines, LS180, HT29, HT29/219, and SW742 by RT-PCR. Furthermore, CEA release could be activated and inhibited by incubation of LS180 cells with suramin and 1,10-phenanthroline, compounds known to activate and inhibit GPI-PLD activity, respectively. The results suggest a mechanism for the involvement of an endogenous GPI-PLD in spontaneous CEA release from human colon cancer cells. PMID- 15893416 TI - The tyrosine kinase inhibitor imatinib fails to inhibit pancreatic cancer progression. AB - Imatinib targets KIT and platelet-derived growth factor receptors (PDGFR) and is highly effective in the treatment of CML and GIST patients. Pancreatic cancers express KIT and PDGFRs. Therefore, 26 patients with unresectable pancreatic cancer were randomized to either gemcitabine (1000 mg/m2 weekly) or imatinib (2x400 mg po) treatment daily. Pancreatic adenocarcinoma was confirmed histologically and expression of KIT and PDGFRbeta was determined immunohistochemically in the biopsy specimens. Quality of life was assessed with two standard questionnaires. No objective responses were seen in either group. Median time to progression was 77 and 29 days (P=0.411) and median survival time was 140 and 60 days (P=0.517) for gemcitabine and imatinib, respectively. Survival and treatment responses were independent of KIT and PDGFRbeta expression in patients treated with imatinib. Grade 3/4 toxicities of imatinib treatment were anemia, elevated liver enzymes, vomiting, and dyspnea. Patients treated with imatinib reported diarrhoea and/or altered bowel function more frequently, which were treatable symptomatically. Quality of life was similar in both groups. In this small series of pancreatic cancer patients, treatment with imatinib was not associated with a significant control of cancer progression. PMID- 15893417 TI - Dose dependent inhibitory effects of dietary 8-methoxypsoralen on NNK-induced lung tumorigenesis in female A/J mice. AB - We have reported that pretreatment by stomach tube with 8-methoxypsoralen (methoxsalen; 8-MOP), a potent human CYP2A6 inhibitor, strongly suppresses lung tumorigenesis by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in female A/J mice (Cancer Res. 2003). Here, we examined inhibitory effects with administration in the diet. When the mice were 7 weeks of age, they received dietary supplementation with 8-MOP at concentrations of 1, 10 or 100 ppm for 3 days prior to a single dose of NNK (2mg/0.1 ml saline/mouse, i.p.) or an equal volume of saline (vehicle control). The experiment was terminated 16 weeks after the first 8-MOP treatment and lung proliferative lesions were analyzed. The incidences and multiplicities in the 8-MOP 100 ppm-treated group were significantly reduced as compared with values for the NNK alone group (P<0.001). Multiplicities of NNK-induced lung proliferative lesions were also reduced in a dose dependent manner (Spearman rank correlation coefficient; rho=-0.806, correction P<0.0001). Mouse CYP2A4 and CYP2A5 differ from each other only 11 amino acids, and are closely related to the human CYP2A6. One hour after the last of three daily doses of 8-MOP (0.5, 5 or 50mg/kg body weight in 0.2 ml corn oil, given by stomach tube) or an equal volume of corn oil (vehicle control), given to the mice at 7 weeks of age, isolation of lung and liver RNAs demonstrated no effects on CYP2A4 and CYP2A5 mRNA levels with 8-MOP. In conclusion, the results of this study showed that clear dose response inhibitory effects of 8-MOP on NNK induced lung tumorigenesis in female A/J mice fed diets containing 8-MOP, due to inhibition of enzyme activity of CYP2A4 and CYP2A5, rather than their gene expression. PMID- 15893418 TI - Synergistic effects of protein tyrosine kinase inhibitor genistein with camptothecins against three cell lines in vitro. AB - Genistein, a natural isoflavone product has been shown to induce cellular death and increase the apoptotic cell death induced by several DNA-damaging stimuli. We have explored the combined effect of genistein and camptothecins against three cell lines, HeLa (cervical cancer), OAW-42 (ovarian cancer) and L929 (normal fibroblasts). Combined effect was estimated in 96-well plates using the SRB method and median-effect analysis. Addition of genistein synergistically increased the antiproliferative affect of camptothecins, inhibiting the camptothecin-induced G2/M arrest and increasing the apoptotic cell population. In HeLa cells, genistein inhibited CDK1 phosphorylation after irinotecan treatment. Thus, abrogation of the G2/M checkpoint control by genistein may be a useful maneuver to increase cytotoxicity of agents that damage DNA and inhibit cell cycle progression in the G2/M boundary. PMID- 15893419 TI - Fecal Adnab-9 binding as a risk marker for colorectal neoplasia. AB - Adnab-9 binding in colonic tissue and effluent has been associated with an increased risk for colorectal neoplasia. We investigated if fecal binding by Adnab-9 may be used as a marker for colorectal neoplasia. A fecal-Adnab-9 ELISA was performed on samples of 249 patients and colonoscopic pathology results correlated. Fecal Adnab-9 binding was seen in 63% of patients with colorectal neoplasia (59% with colorectal cancer and 83% with adenoma), 33% with inflammatory bowel disease, 0% with hyperplastic polyps and 10% of controls. We conclude that fecal Adnab-9 binding is a promising risk marker for colorectal neoplasia. PMID- 15893420 TI - MAPK regulation and caspase activation are required in DMNQ S-52 induced apoptosis in Lewis lung carcinoma cells. AB - 6-(1-Hydroxyimino-4-methylpentyl)5,8-dimethyoxy 1,4-naphthoquinone S-52 (DMNQ S 52) was reported to have cytotoxic activity against L1210 leukemia cells. In the present study, we investigated the apoptotic mechanism of DMNQ S-52 in vitro and in vivo in murine solid cancer cells. DMNQ S-52 exerted cytotoxicity against Lewis lung carcinoma (LLC) cells (IC50=12.3 microM). DMNQ S-52 increased Annexin V positive cell population in a concentration-dependent manner. DMNQ S-52 also induced apoptosis through caspase-mediated pathway, including activation of caspase-3, cleavage of Poly(ADP-ribose) polymerase (PARP) and decreased expression of Bcl-2 in LLC cells in a time and concentration-dependent fashion. DMNQ S-52 activated the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 as well as abrogated the expression of extracellular signal-regulated kinase (ERK) in a time-dependent manner at 10 microM. Similarly, cell proliferation inhibition by DMNQ S-52 was masked by caspase inhibitor Z-Asp-Glu-Val-Asp fluoromethylketone (Z-VAD-FMK), JNK inhibitor SP600125 and p38 inhibitor SB203580, but not by MEK inhibitor U0126. Furthermore, i.p. administration of DMNQ S-52 at 5 mg/kg resulted in a potent inhibition of the growth of LLC cells implanted on the right flank of C57BL/6 mice compared to untreated control. Immunohistochemical analysis revealed the decreased tumor cell proliferation and increased tumor cell apoptosis in DMNQ S-52 treated tumor sections using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) and proliferation cell nuclear antigen (PCNA). Taken together, these findings demonstrate that DMNQ S-52 may exhibit anti-tumor activity by inducing apoptosis via caspases and mitogen activated protein (MAP) kinase-dependent pathways. PMID- 15893421 TI - Towards novel anti-cancer strategies based on cystatin function. AB - Cystatins have recently emerged as important players in a multitude of physiological and patho-physiological settings that range from cell survival and proliferation, to differentiation, cell signaling and immunomodulation. This group of cysteine protease inhibitors forms a large super-family of proteins composed of one, two, three, and, in some species, more than three cystatin domains. Over the last 20 years or so, members of the cystatin super-family have been primarily explored with respect to their capacity to inhibit intracellular cysteine proteases. Yet, this classical mode of action does not fully explain their remarkably diverse biological functions. Due to the space limitations, the author will discuss here the most recent findings that suggest that some of the single-domain, cytoplasmic and cell-secreted cystatins may play important roles in the promotion or suppression of tumor growth, invasion and metastasis. Based on the present understanding of cystatin function, novel avenues for anti-cancer strategies are proposed. PMID- 15893422 TI - Pathogenesis of Lyme neuroborreliosis: mitogen-activated protein kinases Erk1, Erk2, and p38 in the response of astrocytes to Borrelia burgdorferi lipoproteins. AB - Lyme borreliosis, which is prevalent both in the US and in Europe, is an infectious disease that may cause local inflammation in numerous organs. We have hypothesized that, as with some neurodegenerative diseases, the pathogenesis of the neurocognitive deficiencies associated with Lyme neuroborreliosis of the central nervous system also has an inflammatory component. Dysregulated production of pro-inflammatory cytokines such as IL-6 and TNF-alpha can lead to neuronal damage. Mitogen-activated protein kinases (MAPK) play a key role in the regulation of neuronal development, growth, and survival, as well as that of pro inflammatory cytokine production. As a model, we explored the possibility that MAPK-mediated lipoprotein-induced apoptosis and gliosis of rhesus monkey astrocytes stimulated in vitro. Lipoproteins are the key inflammatory molecule type of Borrelia burgdorferi, the spirochete that causes Lyme disease, and we had previously shown that lipoprotein-induced TNF-alpha production in astrocytes caused astrocyte apoptosis, and IL-6 enhanced proliferation of these cells. Lipoproteins readily activated p38 and Erk1/2 MAPK, thus enlisting these pathways among the kinase pathways that spirochetes may address as they invade the central nervous system. We also investigated whether specific inhibition of p38 and Erk1/2 MAPK would inhibit TNF-alpha and IL-6 production and thus astrocyte apoptosis, and proliferation, respectively. Lipoprotein-stimulated IL-6 production was unaffected by the MAPK inhibitors. In contrast, inhibition of both p38 and Erk1/2 significantly diminished TNF-alpha production, and totally abrogated production of this cytokine when both MAPK pathways were inhibited simultaneously. MAPK inhibition thus may be considered as a strategy to control inflammation and apoptosis in Lyme neuroborreliosis. PMID- 15893423 TI - CYP1A1 and CYP1B1 expressions in medulloblastoma cells are AhR-independent and have no direct link with resveratrol-induced differentiation and apoptosis. AB - Resveratrol induces apoptosis and regulates CYP1A1 and CYP1B1 expression in human medulloblastoma cells. To elucidate the potential correlation of their expressions with the anti-medulloblastoma effects of resveratrol, human medulloblastoma cells, UW228-3, were treated with CYP1A1 selective inhibitor (alpha-naphthoflavone, alpha-NF), selective CYP1A1/1A2 inducer (beta naphthoflavone, beta-NF) and their combination with resveratrol, respectively. The influences of those treatments on the expressions of CYP1A1, 1A2 and 1B1 as well as the cell growth, differentiation and death were analyzed. It was found that neither alpha-NF nor beta-NF had any effect on cell growth. alpha-NF inhibited resveratrol-induced CYP1A1 expression without interfering cell differentiation and apoptosis. beta-NF could up-regulate resveratrol-induced CYP1A1 expression but not enhance the anti-cancer effects of resveratrol. CYP1A2 was undetectable in the cells irrespective to the treatments. Aryl hydrocarbon receptor (AhR) was absent in UW228-3 cells under normal culture and treated with resveratrol but induced by both alpha- and beta-NF. Immunohistochemical examination performed on 11 pairs of human medulloblastoma and noncancerous cerebellar tissues revealed that AhR was undetectable in either of them, whereas CYP1A1 was expressed in cerebellum but down-regulated or diminished in their malignant counterparts. Our data suggest for the first time that CYP1A1 and 1B1 expressions in human medulloblastoma cells are AhR-independent and have no direct links with resveratrol-induced differentiation and apoptosis. Appearance of CYP1A1 expression may reflect a more maturated status and a better prognosis of medulloblastomas. PMID- 15893424 TI - Enhancement of wind-up by the combined administration of adenosine A1 receptor ligands on spinalized rats with carrageenan-induced inflammation. AB - The adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA) is very effective in reducing wind-up in intact but not in spinalized adult rats with carrageenan induced inflammation, suggesting an adenosine-mediated supraspinal modulation. Since wind-up is a spinal cord mediated phenomenon but highly influenced by descending modulatory systems, especially in situations of sensitization, we assessed the possible involvement of adenosine in the modulation of wind-up. We studied the effect of the adenosine A(1) receptor antagonist 8-cyclopentyl-1,3 dimethylxanthine (CPT) in the presence and in the absence of the adenosine A(1) receptor agonist CPA. The experiments were carried out in spinalized male Wistar rats under alpha-chloralose anaesthesia. Withdrawal reflexes, studied as single motor units, were activated by noxious mechanical and high-intensity repetitive electrical stimulation (wind-up). While CPA and CPT were not able to induce any change on wind-up when injected alone, the combination of the two drugs, in any order, lead to an important enhancement of wind-up. This enhancement not always paralleled an increase of responses to noxious mechanical stimulation, indicating that the effect is mainly located in the spinal cord. In addition, the enhancement of wind-up was not further increased by the administration of the opioid receptor antagonist naloxone. We conclude that the depression of the wind up phenomenon observed in spinalized animals is, at least in part, dependent of adenosine systems and can be relieved by the combined administration of CPA and CPT. PMID- 15893425 TI - Identifying visual-vestibular contributions during target-directed locomotion. AB - The purpose of this experiment was to examine the potential interaction between visual and vestibular inputs as participants walked towards 1 of 3 targets located on a barrier 5m away. Visual and vestibular inputs were perturbed with displacing prisms and galvanic vestibular stimulation (GVS), respectively. For each target there were three vision conditions (no prisms, prisms left, and prisms right), and three GVS conditions (no GVS, anode left, and anode right). Participants were instructed to start with eyes closed, and to open the eyes at heel contact of the first step. GVS and target illumination were triggered by the first heel contact. This ensured that the upcoming visual condition and target were unknown and that both sensory perturbations occurred simultaneously. Lateral displacement was determined every 40 cm. Irrespective of target or direction, GVS or prism perturbation alone resulted in similar lateral deviations. When combined, the GVS and prism perturbations that had similar singular effects led to significantly larger deviations in the direction of the perturbations. The deviations were approximately equal to the sum of the single deviations indicating that the combined effects were additive. Conflicting GVS and prism perturbations led to significantly smaller deviations that were close to zero, indicating that opposite perturbations cancelled each other. These results show that when both visual and vestibular information remain important during task performance, the nervous system integrates the inputs equally. PMID- 15893426 TI - Effect of acupuncture on 6-hydroxydopamine-induced nigrostratal dopaminergic neuronal cell death in rats. AB - In this study, we investigated the effect of acupuncture at the Zusanli acupoint (ST36) on the nigrostriatal dopaminergic neuronal cell death in the rats with Parkinson's disease. Two weeks after unilateral injection of 6-hydroxydopamine (6 OHDA) into the striatum, an apomorphine-induced rotational behavior test showed significant rotational asymmetry in the rats with Parkinson's disease. Immunostaining for tyrosine hydroxylase demonstrated a dopaminergic neuronal loss in the substantia nigra and dopaminergic fiber loss in the striatum. Acupuncture at the ST36 for 14 days significantly inhibited rotational asymmetry in the rats with Parkinson's disease, and also protected against 6-OHDA-induced nigrostriatal dopaminergic neuronal loss. These effects of acupuncture were not observed for the non-acupoint (hip) acupuncture. The present study shows that acupuncture at the ST36 acupoint can be used as a useful strategy for the treatment of Parkinson's disease. PMID- 15893427 TI - Electroacupuncture prevents cognitive deficits in pilocarpine-epileptic rats. AB - Here we investigated the effects of electroacupuncture over the cognitive deficits in the pilocarpine model of epilepsy in rats. Acupuncture stimulation was provided at acupoints located in either the midline of the back and of the head [HD]: Gv-20 (Baihui), Gv-14 (Dazhui), Gv-2 (Yaoshu) and M-HN-3 (Yin Tang); or acupoints located in the limbs [LB]: St-36 (Zusanli) and Sp-6 (Sanyinjiao). In the elevated T-maze test, electroacupuncture at HD and LB acupoints produced an improvement in the acquisition and retention parameters. Retention in the inhibitory avoidance test was seen only in short-term retention and only for animals stimulated at HD. At histology it was found that electroacupuncture at HD acupoints abolished tissue shrinkage in dorsal hippocampus, basolateral nucleus of the amygdala, substantia nigra and perirhinal cortex, whereas stimulation of LB acupoints prevented tissue shrinkage in all of the above structures except dorsal hippocampus. Administration of p-chlorophenylalanine, a serotonergic releaser, abolished both behavioral and part of the histological changes in these animals. We conclude that electroacupuncture at HD and LB acupoints prevents atrophy of some limbic structures and improves cognitive deficits in pilocarpine epileptic rats and that this effect is dependent on the serotonergic system. PMID- 15893428 TI - Fractal dimension of cerebral cortical surface in schizophrenia and obsessive compulsive disorder. AB - Schizophrenia and obsessive-compulsive disorder (OCD) are assumed to be neurodevelopmental disorders. To examine the cortical patterns in the two disorders, three-dimensional fractal dimension (FD) of skeletonized cerebral cortical surface was estimated from magnetic resonance (MR) images of 50 patients with schizophrenia, 45 patients with OCD and 26 healthy normal controls. The schizophrenic group had a significantly smaller mean FD than OCD group, and the OCD group than normal controls. The FD revealed a significant interaction effect of group-by-hemisphere, and the FD asymmetry index distinguished the schizophrenic group from normal controls. In logistic regression models, the FD and CSF volume correctly classified 95.6% of the schizophrenics from the controls and 88.0% of the patients with OCD from the controls. In the control and schizophrenic groups, the FD was not associated with any of tissue volume measures. In the OCD group, however, the FD was significantly correlated with gray matter tissue volume and intracranial volume (ICV). The results of the present study suggest that three-dimensional FD of cortical surface may be a sensitive indicator for investigation of the structural brain abnormalities in mental disorders, especially those developmentally disturbed. Further studies to explore regional FD changes in mental disorders and clinical implications of the FD including diagnostic value should be performed in the future. PMID- 15893429 TI - The parietal distance effect appears in both the congenitally blind and matched sighted controls in an acoustic number comparison task. AB - Visual world experience is thought to play a significant role in the development of an abstract representation of quantity in the human brain. Nevertheless, some congenitally blind individuals demonstrate excellent numerical abilities. We show that blind adults have a phenomenologically normal semantic representation of number. Electro-encephalography data demonstrate that the numerical distance effect has similar parietal correlates both in the blind and in matched sighted controls. Our interpretation is that number comparison in the blind relies on a compensation network in the initial phase of number comparison. In a second phase, an evolutionarily hardwired parietal system is exploited. The representation of number meaning has both plastic and evolutionarily hardwired components. PMID- 15893430 TI - Aortic thromboembolism associated with Spirocerca lupi infection. AB - A 2-year-old male castrated Cavalier King Charles Spaniel was presented with paraplegia, cold caudal extremities and lack of femoral pulses. A 2cm long thrombus occluding the aortic trifurcation and a 3cm long abdominal aortic aneurysm with a thrombus were detected by ultrasonographic examination. The clinical and ultrasonographic findings were consistent with aortic thromboembolism. Anti-thrombotic and vasodilative therapy was not helpful and the dog was euthanized 3 days after the onset of paraplegia. A thrombus in the aortic trifurcation, multiple thoracic and abdominal aneurysms and a distal mediastinal esophageal granuloma containing Spirocera lupi worms were found on necropsy. The abdominal aortic aneurysms formed by S. lupi larval migration are believed to be responsible for the formation of the thrombus that occluded the aortic trifurcation. This is the first report of aortic thromboembolism associated with S. lupi infection. PMID- 15893431 TI - Efficacy of a combination of imidacloprid 10%/permethrin 50% versus fipronil 10%/(S)-methoprene 12%, against ticks in naturally infected dogs. AB - Preventing tick bites is a fundamental step towards reducing the impact of tick borne protozoal, bacterial and viral diseases (TBDs) in humans and animals. The aim of this study was to evaluate the efficacy of a combination of imidacloprid 10%/permethrin 50% and of fipronil 10%/S-methoprene 12% against ticks in naturally infected dogs and to assess methodological parameters to calculate drug efficacy on tick immature stages. From July to August 2004, 45 privately owned dogs of various sexes, ages, breeds, coat length and habits were enrolled in a trial carried out in an area (radius approximately 50km) in Southern Italy. Three homogeneous groups (both for dog population and tick population) were formed: 15 dogs treated with imidacloprid 10% and permethrin 50% spot-on (group A), 15 dogs treated with fipronil 10% and methoprene 12% spot-on (group B) and 15 untreated dogs (group C). The dogs in each group were then sub-grouped according to their age and weight. Two different treatments were administered (time 0 and +28 days) to groups A and B, and the dogs were checked weekly for tick infestation until day +56 post-treatment (p.t.). Twenty-four areas distributed on the whole body surface were examined for ticks at each follow-up, while only at time 0 and at day +56 p.t., ticks were collected from the dogs and identified. For the immature stages a semi-quantitative method was adopted and the load of immature stages was evaluated and grouped into four classes up to day +56 p.t. when the mean number of immature ticks (MIT) for each infection class was evaluated. All the adult ticks collected were identified as brown dog ticks (Rhipicephalus sanguineus). Immature stages were first compared at day +28 p.t.. The efficacy of both products used in groups A and B on adult ticks was high and generally very similar. Conversely, the efficacy of imidacloprid 10% and permethrin 50% against immatures was higher than that of fipronil 10% and methoprene 12% throughout the observation period with statistically significant differences (p<0.05) at day +28 p.t. (i.e. group A=98.52%, group B=72.40%). On the whole, in analysing the efficacy of both products against adult plus immature ticks, it was found that the combination of imidacloprid 10% and permethrin 50% was more effective than fipronil 10% and methoprene 12%, with the differences being statistically significant at day +28 p.t. (group A=98.43%, group B=77.56%). PMID- 15893432 TI - Subthalamic nucleus lesion reverses motor abnormalities and striatal glutamatergic overactivity in experimental parkinsonism. AB - Subthalamic nucleus (STN) is a target of choice for the neurosurgical treatment of Parkinson's disease (PD). The therapeutic effect of STN lesion in PD is classically ascribed to the rescue of physiological activity in the output structures of the basal ganglia, and little is known about the possible involvement of the striatum. In the present study, therefore, we electrophysiologically recorded in vitro single striatal neurons of DA-depleted rats unilaterally lesioned by 6-hydroxydopamine, treated or not with therapeutic doses of levodopa (l-DOPA), or with a consecutive ipsilateral STN lesion. We show that the beneficial motor effects produced in parkinsonian rats by STN lesion or l-DOPA therapy were paralleled by the normalization of overactive frequency and amplitude of striatal glutamate-mediated spontaneous excitatory postsynaptic currents (sEPSCs). Since neither l-DOPA treatment nor STN lesion affected sEPSCs kinetic properties, the reversal of these abnormalities in striatal excitatory synaptic transmission can be attributable to the normalization of glutamate release. PMID- 15893433 TI - Internet and paper self-help materials for problem drinking: is there an additive effect? AB - The objective of this study was to conduct a preliminary evaluation of an Internet-based intervention for problem drinkers, comparing changes in drinking between respondents who only received the intervention to those who also received a self-help book. After receiving a personalized feedback summary on the Internet, 83 respondents provided complete baseline information and volunteered to participate in a 3-month follow-up survey. Half of the respondents were randomized to receive an additional self-help book. The follow-up was returned by 48 respondents (69% female). Repeated measures ANOVAs were conducted to compare drinking levels at baseline and 3-month follow-up among respondents who only received the Internet-based intervention. There was minimal support for an impact of the Internet intervention alone. In addition, hierarchical regression analyses were conducted to compare respondents in the two intervention conditions on their drinking at follow-up, controlling for baseline consumption. Respondents who received the additional self-help book reported drinking less and experiencing fewer consequences at follow-up as compared to respondents who received only the Internet-based intervention. While the results are promising, they cannot be taken as evidence of the efficacy of Internet-based personalized feedback as a stand-alone intervention because of the absence of a control group that did not receive the intervention. Further research on this topic should be a priority because of the potential for Internet-based interventions to reach problem drinkers underserved by traditional treatment. PMID- 15893434 TI - Lignans from Schisandra hernyi with DNA cleaving activity and cytotoxic effect on leukemia and Hela cells in vitro. AB - The lignans, gomisin G (1), schisantherin A (2), benzoylgomisin Q (3) and isoanwulignan (4) were isolated from the stems of Schisandra henryi. Compound 1 showed moderate DNA strand scission activity and significant cytotoxic effect on leukemia and Hela cells in vitro. Compound 1 represents a new type of DNA strand scission agent. PMID- 15893435 TI - HPLC based activity profiling for 5-lipoxygenase inhibitory activity in Isatis tinctoria leaf extracts. AB - In the pursuit of the anti-inflammatory constituents in lipophilic woad extracts, the 5-lipoxygenase (5-LOX) inhibitory activity was investigated by HPLC-based activity profiling. In a low-resolution profiling, two time windows with peaks of activity were found. The first coincided with tryptanthrin, a known dual inhibitor of cyclooxygenase-2 (COX-2) and 5-LOX, whereas the major inhibitory fraction was towards the end of the HPLC run. The active fractions were profiled in a peak-resolved manner, and the compounds analyzed by LC-MS, GC and TLC. The activity in the lipophilic fractions of the Isatis extract could be linked to an unsaturated fatty acid, alpha-linolenic acid. PMID- 15893436 TI - Characterization of a ribonuclease gene and encoded protein from the reptile, Iguana iguana. AB - In this work we identify an intronless open reading frame encoding an RNase A ribonuclease from genomic DNA from the Iguana iguana IgH2 cell line. The iguana RNase is expressed primarily in pancreas, and represents the majority of the specific enzymatic activity in this tissue. The encoded sequence shares many features with its better-known mammalian counterparts including the crucial His12, Lys40 and His114 catalytic residues and efficient hydrolytic activity against yeast tRNA substrate (k(cat)/K(m)=6 x 10(4) M(-1) s(-1)), albeit at a reduced pH optimum (pH 6.0). Although the catalytic activity of the iguana RNase is not diminished by human placental RI, iguana RNase is not bactericidal nor is it cytotoxic even at micromolar concentrations. Phylogenetic analysis indicates moderate (46%) amino acid sequence similarity to a pancreatic RNase isolated from Chelydra serpentina (snapping turtle) although no specific relationship could be determined between these RNases and the pancreatic ribonucleases characterized among mammalian species. Further analysis of ribonucleases from non-mammalian vertebrate species is needed in order to define relationships and lineages within the larger RNase A gene superfamily. PMID- 15893437 TI - Genomic organization and promoter analysis of the mouse ADP-ribosylarginine hydrolase gene. AB - Mono-ADP-ribosylation is a reversible modification of proteins with NAD:arginine ADP-ribosyltransferases and ADP-ribosylarginine hydrolases (ADPRH) catalyzing the opposing arms of an ADP-ribosylation cycle. The ADPRH cDNA had been cloned from human, rat, and mouse tissues and high levels of mRNA were found in brain, spleen, and testis. To begin to understand the molecular mechanisms that regulate ADPRH gene expression, we cloned the full-length cDNA, determined the genomic structure of mouse ADPRH, and investigated promoter function. Northern analysis using different regions of the ADPRH cDNA as probes identified two mRNAs of 1.7 and 3.0 kb, which resulted from the use of alternative polyadenylation signals, CATAAC and ATTAAA, beginning at positions 1501 and 2885, respectively, of the nucleotide sequence (A of ATG = 1). The ADPRH gene, represented in two overlapping genomic clones, spans approximately 9 kilobases with four exons and three introns. The 5'-flanking region contains the features of a housekeeping gene; it has neither a TATA nor a CAAT box, but is, instead, highly GC-rich with multiple transcription initiation sites. Promoter analysis, assessed using transient transfection of PC12, NB41A3, NIH/3T3, and Hepa 1-6 cells with truncated constructs, revealed potent stimulatory (-119 to -89) and inhibitory ( 161 to -119) elements, which were utilized similarly in the different cell lines. Further mutational analysis of the promoter and electrophoretic mobility-shift assays identified a positive GC-box element (-107 to -95); Sp1 and Sp3, which bound to this motif, were also detected by supershift assays. In co-transfection experiments using Drosophila SL2 cells that lack endogenous Sp1, Sp1 trans activated the ADPRH promoter in a manner dependent on the presence of an Sp1 binding motif. The promoter activity pattern and involvement of Sp transcription factors are consistent with prior observations of widespread hydrolase expression in mammalian tissues. PMID- 15893438 TI - Roles of plasma proteins in the formation of silicotic nodules in rats. AB - The contribution of plasma protein(s) to the stabilization of fibroids formed in rat lungs exposed to acute silica dust inhalation was examined. Antibodies against component proteins of the nodules remaining insoluble in 2% SDS, 10M urea and 40 mM sulfhydryl reagents under prolonged boiling conditions were raised in rabbits and used to capture plasma proteins, which were identified by 2D-gel electrophoresis and MALDI-TOF analysis. The silica particles were encapsulated with extracellular protein composites whose amino acid compositions showed high levels of alanine, i.e., above those of glycine and proline, a building block of collagen. Antibody-captured plasma proteins showed the dominant presences of fibrinogen, albumin, and prealbumin (transthyretin), and other minor proteins, which included alpha-1-protease inhibitor, contraspin-like protease inhibitor, cathepsin B, etc. The presence of the N(epsilon)-(gamma-glutamyl) lysine isopeptide bond in the nodules was evidenced by direct chemical methods and by immunoreactivity for anti-isopeptide bonds. Immunostaining of affected lung tissue and of the fibroid regions showed elevated levels of transglutaminase (TGase) E and plasma factor XIII (F-XIII), but showed no reactivity towards other TGases. These findings suggest that the silica encapsulated nodules are a mixture of extracellular proteins that include collagen type I, fibrin and transthyretin, which is stabilized by TGase catalyzed crosslinking between plasma and extracellular proteins during fibrosis to eventually form insoluble nodules. PMID- 15893439 TI - Preparation and in vitro evaluation of chitosan nanoparticles containing a caspase inhibitor. AB - The aim of this work was to develop a formulation for Z-DEVD-FMK, a peptide which is a caspase inhibitor and has been used in experimental animal studies for a decade. Peptide loaded chitosan nanoparticles were obtained by ionotropic gelation process and Z-DEVD-FMK was quantified by an HPLC method. The influence of the initial peptide concentration on the nanoparticle characteristics and release behavior was evaluated. The CS nanoparticles have a particle diameter (Z average) ranging from approximately 313-412 nm and a positive zeta potential (20 28 mV). The formulation with the initial peptide concentration of 400 ng/ml provided the highest loading capacity (0.46%) and the highest extent of release (65% at 24 h) suggesting the possibility to achieve a therapeutic dose. According to the data obtained, this chitosan-based nanotechnology opens new and interesting perspectives for anticaspase activity. PMID- 15893440 TI - Effective comparison of two auto-CPAP devices for treatment of obstructive sleep apnea based on polysomnographic evaluation. AB - BACKGROUND: Automatic continuous positive airway pressure (auto-CPAP) machines differ mainly in algorithms used for respiratory event detection and pressure control. The auto-CPAP machines operated by novel algorithms are expected to have better performance than the earlier ones in the treatment of obstructive sleep apnea syndrome (OSAS). OBJECTIVES: The purpose of this study was to determine the therapeutic characteristics between two different auto-CPAP devices, i.e., the third-generation flow-based (f-APAP) and the second-generation vibration-based (v APAP) machines, during the first night treatment of OSAS. METHODS: We retrospectively reviewed the polysomnography (PSG) recordings of 43 OSAS patients who were initially performed an overnight diagnostic PSG to confirm the disease and afterwards received the first night auto-CPAP treatment with using either the f-APAP (n=22) or v-APAP (n=21) device under another PSG evaluation. RESULTS: There were 13.6% and 61.9% patients who remained a residual apnea/hypopnea index more than 5 during the f-APAP and v-APAP application, respectively (P<0.005). The f-APAP was more effective than the v-APAP in reducing apnea/hypopnea index (P=0.003), hypopnea index (P=0.023) and apnea index (P=0.007), improving the lowest oxygen saturation index (P=0.007) and shortening stage 1 sleep (P=0.016). However, the f-APAP was less sufficient than the v-APAP in reducing arousal/awakening index (P=0.02). CONCLUSION: These findings suggest that the f APAP works better than the v-APAP in abolishing breathing abnormities in the treatment of OSAS; however, the f-APAP device might still have some potential limitations in the clinical application. PMID- 15893441 TI - The effects of section thickness on the estimation of liver volume by the Cavalieri principle using computed tomography images. AB - Estimation of liver volume using routine CT scans has been described previously. We have, however, not found a gold standard study which analyzes the effect of section thickness on the estimation of liver volume using CT images. In the present study, five normal livers obtained from cadavers were scanned using a Spiral CT Scanner (Xpress/GX Toshiba, Tocigi-Ken) in the horizontal plane. Consecutive sections at a thickness of 10, 5 and 1mm were used to estimate the total volume of the livers by means of the Cavalieri principle. With a view to evaluating inter-observer differences, liver volume was estimated by three observers. The estimated volume using the classical volume estimation formula did not concur with the actual volume of the livers obtained by the fluid displacement technique. The section thickness has an over- or under-projection effect on the estimated volume. The obtained volume estimation results were, therefore, calibrated using three different approaches. The volume obtained by the calibration formulae did not differ statistically from actual liver volumes (P<0.05). There were also no significant differences between the performers' estimates (P>0.05). Results showed that the effect of section thickness on the volume estimates could not be omitted and the obtained values could be calibrated using the proposed approaches presented in this study. PMID- 15893442 TI - Validation of quantitative NMR. AB - NMR is by definition a quantitative spectroscopic tool because the intensity of a resonance line is directly proportional to the number of resonant nuclei (spins). This fact enables, in principle, a precise determination of the amount of molecular structures and, hence, of substances in solids as well as liquids. With the increase of sensitivity due to stronger and stronger static magnetic fields including improved electronics the detection limits have been pushed down significantly. However, the lack of a precise protocol that considers and controls the aspects of both the measurement procedure as well as the spectra processing and evaluation is responsible for the fact that quantitative investigations of identical samples in various laboratories may differ severely (deviations up to 90% relative to gravimetric reference values). Here, a validated protocol for quantitative high resolution 1H-NMR using single pulse excitation is described that has been confirmed by national and international round robin tests. It considers all issues regarding linearity, robustness, specificity, selectivity and accuracy as well as influences of instrument specific parameters and the data processing and evaluation routines. This procedure was tested by the investigation of three different 5-model-compound mixtures. As a result of the round robin tests using the proposed protocol it was found that the maximum combined measurement uncertainty is 1.5% for a confidence interval of 95%. This applies both for the determination of molar ratios and of the amount fractions of the various components. Further, the validation was extended to purity determinations of substances as shown for 1,8-epoxy-p-menthane (cineole). PMID- 15893443 TI - Molecular pathogenetic mechanisms of nephrotic edema: progress in understanding. AB - Molecular and pathogenetic mechanisms in sodium retention and water reabsorption of nephrotic edema are discussed. Are reported and analyzed molecular mechanisms about sodium retention in collecting duct cells regarding activation and surface expression of epithelial sodium channels (ENaC) and sodium-potassium-ATPase (Na,K ATPase) by aldosterone, vasopressin, natriuretic peptide system (underfill theory): is necessary a better understanding about the dysregulation of ENaC and Na,K-ATPase surface expression and the resistance to natriuretic peptide system. Are also reported and analyzed molecular mechanisms of sodium retention in proximal tubule cells regarding intrinsic albumin toxicity upon type 3 sodium hydrogen exchanger ionic pump and the activity of sodium-hydrogen exchanger regulatory factor protein (overfill theory): a better knowledge about the link between albumin, sodium-hydrogen exchanger type 3 (NHE3) ionic pump, sodium hydrogen exchanger regulatory factor protein is necessary. Then molecular mechanisms of vasopressin free water retention through acquaporin water channels in collecting duct cells are discussed: further studies are necessary to understand vasopressin release pathway (osmotic/nonosmotic) and V2 receptor activation with cell surface expression of renal acquaporins water channel. PMID- 15893444 TI - Na+K+-ATPase activity as a biomarker of toxaphene toxicity in Unio tumidus. AB - In this study, the effect of toxaphene (camphechlor) on ATPase activity in the microsomal fraction of the Unio tumidus's digestive gland was determined. Toxaphene is a man-made mixture consisting of polychlorinated monoterpens, predominantly bornanes. This compound was primarily used as an insecticide, but in 1982 was officially banned because of its destructive effects on human and animal health. Toxaphene can be transported in the air at long distances and can persist in air, soil and water for years revealing acute and chronic toxicity towards aquatic organisms and wildlife, the increasing risk of cancer in both humans and animals. The microsomal fraction isolated from digestive glands was exposed to 1 x 10(-3) M, 1 x 10(-5) M and 1 x 10(-7) M of toxaphene. The obtained data showed that toxaphene induced a loss of ATPase activity in all used concentrations. The Lineweaver-Burk plots for microsomal Na+K+-ATPase in the presence or the absence of toxaphene as an inhibitor indicated a competitive type of inhibition. PMID- 15893445 TI - Comparison of basal cytotoxicity data between mammalian and fish cell lines: a literature survey. AB - During the last 20 years, in vitro assays with fish cells have been used in ecotoxicology for testing chemicals, effluents and in toxicity identification evaluations (TIE) studies. Due to the good correlation found between the in vitro data and the in vivo fish data in ranking chemical toxicity, they have been proposed as an alternative to acute fish bioassays for toxicity screening of chemicals. Nevertheless, they are recognized to be less sensitive when compared with in vivo fish bioassays. Fish cells have slower cell cycles than mammalian cells, so it was suggested that mammalian cells could be more sensitive than fish cells for testing chemicals. However, as fish cells offer some advantages over mammalian cells, mainly related to their ease of handling, some authors found them preferable to use for routine testing. The present study was undertaken to analyze whether mammalian cell lines are more, less, or equally sensitive, compared to fish cells lines. To this end, basal cytotoxicity data from a literature survey of mammalian and fish cell lines have been compared. There was a good linear correlation of IC50 values (r = 0.915) between fish and mammalian cells for a set of 51 chemicals after 24-h of treatment. The correlation coefficient of IC50 values (r = 0.888) decreased when 24-h treated mammalian cells were correlated against 48-h treated fish cell lines. For the dataset selected for this study, fish and mammalian cells showed a similar sensitivity for most of the chemicals after being treated for 24 h. We conclude that fish and mammalian cells can equally well predict the basal toxicity of the set of chemicals used in this study, and that increased exposure periods did not increase the correlation between midpoint toxicities nor sensitivities for any of the cell lines used. The more practical handling of fish cells favors their recommendation as a better alternative for replacing fish bioassay than mammalian cells. PMID- 15893446 TI - In vitro cytotoxicity of selected chemicals commonly produced during fire combustion using human cell lines. AB - Fire combustion products contain a broad range of chemicals, which have a multitude of possible toxic interactions in humans. The aim of this study was to evaluate the cytotoxicity of selected substances commonly produced during fire combustion. A range of human cell lines and cultures including: skin fibroblasts, HepG2 (liver derived), and A549 (lung derived cells) were used to represent different human target organs. The colorimetric MTS assay (3-(4,5-dimethylthiazol 2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) was used to detect the cytotoxic effects of selected substances including: acetic acid, ammonia, formaldehyde, hydrobromic acid, hydrochloric acid, hydrofluoric acid, potassium cyanide, sodium fluoride, sodium nitrite, sodium sulphide, and sulphurous acid. In this study, the NOAEC (No Observable Adverse Effect Concentration), IC(10) (10% inhibitory concentration), IC50 (50% inhibitory concentration), and TLC (Total Lethal Concentration) values were determined. The ratio between in vitro IC50 to in vivo human toxicity data (Lowest Lethal Dose LDLo and Lowest Lethal Concentration--LCLo) was also established. Results indicated a strong relationship between IC50 values on the cell types used: fibroblast and A549 (R2: 0.92), A549 and HepG2 (R2: 0.72), fibroblast and HepG2 (R2: 0.69). Good correlation was obtained between the IC50 against LDLo and LCLo when an appropriate adjustment factor was implemented. Results of this study indicated that in vitro methods could be a potential technique for assessing the toxicity of fire combustion products. PMID- 15893447 TI - Screening of biological activities present in honeybee (Apis mellifera) royal jelly. AB - Bioactivities present in three honeybee royal jelly (RJ) protein fractions were screened for diverse in vitro model systems. RJCP, a RJ crude protein extract, stimulated cell growth of Tn-5B1-4 insect cells inducing 6.5 population doublings per mg of protein added to culture medium, meanwhile fetal bovine serum, the usual growth supplement, gave rise only to 2.55 population doublings. RJCP, as well as the fractions RJP30 and RJP60 obtained by precipitation with ammonium sulfate, also affected Tn-5B1-4 cell shape and stimulated adhesion of these cells to the substrate. RJP30 also increased the percent of mature adipocytes in cultures of 3T3-F442A murine preadipocytes twofold with respect to insulin treatment, without inducing additional cell growth of confluent preadipocytes. Fractions RJCP and RJP60 showed similar capacity as that of insulin to stimulate the formation of mature 3T3 adipocytes. Fraction RJP30 also was cytotoxic for HeLa human cervicouterine carcinoma cells, diminishing 2.5-fold the initial cell density after seven days of treatment. Our results show the presence of diverse bioactivities in RJ affecting cell growth, cell differentiation and cell survival of insect, murine, and human cancer cells. PMID- 15893448 TI - In vitro models to evaluate acute and chronic injury to the heart and vascular systems. AB - Multiple in vitro model systems are currently available to evaluate structure and function relationships in the cardiovascular system as well as the system's response to injury. As the level of molecular sophistication continues to advance, so does the level of complexity of the analysis. One of the most daunting tasks faced by researchers interested in studying cardiovascular function and injury is the selection of the system or systems best suited to answer the particular question at hand. In order to successfully apply any given model system, the researcher must recognize the advantages and limitations in the system of choice. This review provides a listing of the historical and modern techniques used to study cardiovascular function and chemically-induced toxicity. With the growing number of new pharmaceuticals discovered each year, it is imperative to use experimental model systems that allow for identification of targets that participate in or mediate adverse outcomes. Clearly, in vitro analysis cannot replace in vivo experimentation, but the methods currently available allow for a reduction in the number of animals used for experimentation and a better understanding of the complexity associated with the injury response. PMID- 15893449 TI - Substrate specific metabolism by polymorphic cytochrome P450 2D6 alleles. AB - A comparative metabolism study was performed for bufuralol, dextromethorphan, imipramine, mianserin, sparteine, tamoxifen, haloperidol and two drug candidates (Rec27/0110 and Rec15/2739) on V79 cells genetically engineered to express human cytochrome P450 (CYP) variants 2D6*1, 2D*2, 2D*9 and 2D*17. Unexpectedly, the CYP2D6*17 dependent metabolism profile of haloperidol and Rec27/0110 were found to differ from all other substrates tested. Some of these known standard substrates are frequently applied in marker reactions for CYP2D6 and with these standard substrates, CYP2D6*1 is known to be the most active form. In both cases of haloperidol and Rec27/0110 the variant form CYP2D6*17 had equal or higher activity compared to the CYP2D6*1 form. Results obtained with the V79 cells were confirmed using microsomal preparation of yeast cells expressing the variants CYP2D6*1 and CYP2D6*17 and CYP2D6 inhibitor quinidine. In conclusion, there is no general rule for a variant dependent metabolism profile by cytochrome P450 2D6 indicating that the activity profile of the CYP2D6 alleles may be substrate specific, thus may be reflected in pharmacokinetics consequences for individuals. PMID- 15893451 TI - Reconstruction and exploration of three-dimensional confocal microscopy data in an immersive virtual environment. AB - An immersive virtual environment for interactive three-dimensional reconstruction and exploration of confocal microscopy data is presented. For some structures automatic alignment of serial sections can lead to geometric distortions. The superior visual feedback of a Virtual Reality system is used to aid in registering and aligning serial sections interactively. An ImmersaDesk Virtual Reality display system is used for display and interaction with the volumetric confocal data. Detailed methods for handling both single-section and multi section confocal data are described. PMID- 15893452 TI - Protecting patient privacy against unauthorized release of medical images in a group communication environment. AB - In this paper, we identify and study an important patient privacy protection problem related to medical images. Following Health Insurance Portability and Accountability Act (HIPAA) mandate on privacy protection of patients' medical records, efforts have been devoted to guaranteeing the confidentiality of data and medical images during storage and transmission via an untrustworthy channel. However, to our knowledge, there has not been any effort towards protecting against unauthorized release of images by an authorized recipient. In this paper, we study the problem of tracing illegally distributed medical images in a group communication environment and identify a set of design requirements that must be met. We propose a fingerprint model suitable for many-to-many multicast, that is computationally efficient and scalable in user storage and key update communication. Simulation results also show that our scheme is highly robust to typical medical image processing and collusion attacks, while yielding high quality watermarked images. PMID- 15893453 TI - Vessel enhancement in digital X-ray angiographic sequences by temporal statistical learning. AB - In this paper, we present a vessel enhancement method, SVM temporal filtering (STF), for X-ray angiographic (XA) images using Support Vector Machine (SVM). We show that the linear SVM applied to vessel enhancement can be regarded as a matched linear filter optimizing the contrast-to-noise ratio in XA images. We propose a non-linear kernel function for the SVM leading to good enhancement with noisy, varying grey-level dynamics at vessel pixels. One key advantage over the matched filters is that an optimal filter is learnt from images, not estimated at design stage. Results on clinical XA images show that learning-based enhancement achieves better results compared to simple subtraction and other image stacking methods. PMID- 15893454 TI - A pixel-based autofocusing technique for digital histologic and cytologic slides. AB - The present paper describes a method for autofocusing specifically studied for the acquisition of digital slides, i.e. full histologic and cytologic slides, utilising low-cost equipment. At first, experimentations with some of the most used focus measures and algorithms have been made, in order to choose the most suitable for histologic and cytologic images. Then, a study of the specific features of digital slides has been preliminarily carried out in order to understand the constraints of the domain. These included the capability of autofocusing in an unattended way on thousands of microscope fields, while fast performance is not a strict requirement. Based on the findings, an algorithm based on a dynamic focus position space, with a variance-based focus measure has been adapted to the specific situation. A qualitative and quantitative evaluation of the proposed algorithm allowed us to show that the proposed algorithm is suitable for the acquisition of digital slides, and furthermore it can be implemented starting from a basic microscope with an inexpensive motorised stage. The algorithm is currently implemented into a complete digital slide acquisition system, which is in turn being used for a Quality Assurance Programme in cervicovaginal cytologic screening. PMID- 15893455 TI - The conditional kinase DeltaMEKK1:ER* selectively activates the JNK pathway and protects against serum withdrawal-induced cell death. AB - The conditional protein kinase DeltaMEKK3:ER* allows activation of the mitogen activated and stress-activated protein kinases (MAPKs and SAPKs) without imposing a primary cellular stress or damage. Such separation of stress from stress induced signalling is particularly important in the analysis of apoptosis. Activation of DeltaMEKK3:ER* in cycling CCl39 cells caused a rapid stimulation of the ERK1/2, JNK and p38 pathways but resulted in a slow, delayed apoptotic response. Paradoxically, activation of the same pathways inhibited the rapid expression of Bim(EL) and apoptosis following withdrawal of serum. Inhibition of the ERK1/2 pathway prevented the down-regulation of Bim(EL) but caused only a partial reversion of the cyto-protective effect of DeltaMEKK3:ER*. In contrast, inhibition of p38 had no effect, raising the possibility that activation of JNK might also exert a protective effect. To test this we used CCl39 cells expressing DeltaMEKK1:ER* which activates JNK but not ERK1/2, p38, PKB or IkappaB kinase. Activation of DeltaMEKK1:ER* inhibited serum withdrawal-induced conformational changes in Bax and apoptosis. These results suggest that in the absence of any overt cellular damage or chemical stress activation of JNK can act independently of the ERK1/2 or PKB pathways to inhibit serum withdrawal-induced cell death. PMID- 15893456 TI - Nerve growth factor (NGF) and epidermal nerve fibers in atopic dermatitis model NC/Nga mice. PMID- 15893457 TI - Morphology of ferret subcutaneous adipose tissue after 6-month daily supplementation with oral beta-carotene. AB - Adipose tissue is an important retinoid depot and retinoids are known to influence white and brown adipocyte metabolism. Identifying nutrients that can affect the biological activity of the adipose organ would be of great medical interest in the light of the current obesity epidemic and related disorders in developed countries. The vast majority of mammal studies of chronic administration of oral beta-carotene have used murine models, while few have employed mammals exhibiting uptake and processing of intestinal beta-carotene similar to those of humans. While rodents transform practically all ingested beta carotene into retinol, in ferrets, as in humans, part of the beta-carotene is absorbed and released into the circulation intact. We studied the effects of 6 month daily administration of two doses of oral beta-carotene (0.8 or 3.2 mg/kg/day) on ferret body weight, size of body fat depots, and, using morphological and morphometric methods, on subcutaneous (inguinal) white adipose tissue (WAT). Because of the oral mode of administration, liver, stomach, and small and large intestine were also studied. Control animals received the vehicle. Data show that at the end of treatment the higher dose induced significantly higher body weight compared with controls and significantly higher inguinal fat depot compared with animals treated with the lower dose. In addition, chronic treatment with beta-carotene induced a dose-dependent hypertrophy of white adipocytes and increased neoangiogenesis in subcutaneous WAT in all treated ferrets. Vasculogenesis was independent of adipocyte hypertrophy. We also found focally evident liver steatosis in the ferrets treated with the higher dose of beta-carotene. The other gastrointestinal tract organs studied were not significantly different from those of control animals. PMID- 15893458 TI - An fMRI investigation of strategies for word recognition. AB - A common procedure used to study visual word recognition is the lexical decision task (LDT). Behavioral studies have demonstrated that overall performance in this task is modulated by the type of foils presented. There are divergent claims about the impact of different types of foils on overall processing strategies in the LDT: some researchers claim that pseudohomophone foils (e.g., BRANE) encourage participants to selectively de-emphasize phonological processing, while other researchers claim that pseudohomophone foils encourage participants to engage in more extensive processing of all types (orthographic, phonological, and semantic). To evaluate these claims, we conducted a functional magnetic resonance imaging study. Participants (N = 18) completed three lexical decision tasks, each with a different foil type: consonant strings (e.g., BVRNT), pseudowords (e.g., BLINT), and pseudohomophones (e.g., BRANE). We presented homophones (e.g., MAID) and nonhomophones (e.g., MESS) on word trials in order to be able to calculate the homophone effect as a marker of phonological processing for word stimuli in each foil condition. Comparison of behavioral results in the different foil conditions showed that reaction times were longest, error rates were highest, and homophone effects were largest in the LDT with pseudohomophones. Imaging results showed the greatest magnitude of activity in several regions, including the inferior frontal cortex, during the LDT with pseudohomophone foils. A comparison to inferior frontal activity produced during an additional task (rhyme judgment) supported the conclusion that LDT with pseudohomophone foils is a difficult task in which readers engage in an overall response strategy involving extensive processing of phonological information. PMID- 15893459 TI - Role of liquid membrane phenomenon in the biological actions of thioridazine. AB - Role of surface activity in the mechanism of action of thioridazine (THR) has been studied. THR has been shown to generate liquid membrane it self and also in association with the relevant membrane lipids, sphingomyelin and cholesterol in series with a supporting membrane. Transport of relevant biogenic amines e.g. dopamine, nor-adrenaline, adrenaline, serotonin, gamma amino butyric acid (GABA) and glutamic acid and ions viz. sodium, potassium, and calcium has been studied in the presence of liquid membranes generated by THR and THE in association with sphingomyelin-cholesterol. The data on modifications in the permeability of relevant biogenic amines and ions indicate that the liquid membranes generated by THR may contribute to the mechanism of action of THR. PMID- 15893460 TI - Optimisation of the predictive ability of artificial neural network (ANN) models: a comparison of three ANN programs and four classes of training algorithm. AB - The purpose of this study was to determine whether artificial neural network (ANN) programs implementing different backpropagation algorithms and default settings are capable of generating equivalent highly predictive models. Three ANN packages were used: INForm, CAD/Chem and MATLAB. Twenty variants of gradient descent, conjugate gradient, quasi-Newton and Bayesian regularization algorithms were used to train networks containing a single hidden layer of 3-12 nodes. All INForm and CAD/Chem models trained satisfactorily for tensile strength, disintegration time and percentage dissolution at 15, 30, 45 and 60 min. Similarly, acceptable training was obtained for MATLAB models using Bayesian regularization. Training of MATLAB models with other algorithms was erratic. This effect was attributed to a tendency for the MATLAB implementation of the algorithms to attenuate training in local minima of the error surface. Predictive models for tablet capping and friability could not be generated. The most predictive models from each ANN package varied with respect to the optimum network architecture and training algorithm. No significant differences were found in the predictive ability of these models. It is concluded that comparable models are obtainable from different ANN programs provided that both the network architecture and training algorithm are optimised. A broad strategy for optimisation of the predictive ability of an ANN model is proposed. PMID- 15893461 TI - Microencapsulated chitosan nanoparticles for lung protein delivery. AB - It has already been demonstrated that spray drying is a very valuable technique for producing dry powders adequate for pulmonary delivery of drugs. We have developed chitosan/tripolyphosphate nanoparticles that promote peptide absorption across mucosal surfaces. The aim of this work was to microencapsulate protein loaded chitosan nanoparticles using typical aerosol excipients, such as mannitol and lactose, producing microspheres as carriers of protein-loaded nanoparticles to the lung. The results showed that the obtained microspheres are mostly spherical and possess appropriate aerodynamic properties for pulmonary delivery (aerodynamic diameters between 2 and 3 microm, apparent density lower than 0.45 g/cm3). Moreover, microspheres morphology was strongly affected by the content of chitosan nanoparticles. These nanoparticles show a good protein loading capacity (65-80%), providing the release of 75-80% insulin within 15 min, and can be easily recovered from microspheres after contact with an aqueous medium with no significant changes in their size and zeta potential values. Therefore, this work demonstrated that protein-loaded nanoparticles could be successfully incorporated into microspheres with adequate characteristics to reach the deep lung, which after contact with its aqueous environment are expected to be able to release the nanoparticles, and thus, the therapeutic macromolecule. PMID- 15893462 TI - [Usefulness of procalcitonin for the diagnosis of acute pyelonephritis in children]. AB - BACKGROUND: Acute pyelonephritis can induce parenchymal scarring. The aim of this study was to evaluate the usefulness of procalcitonin (PCT) to predict renal involvement in febrile children with urinary tract infection (UTI). METHODS: In a prospective study serum PCT was measured and compared with others commonly used inflammatory markers in children admitted to the emergency unit with acute pyelonephritis. Renal parenchymal involvement was assessed by a (99 m)Tc-labeled dimercaptosuccinic acid (DMSA) renal scar performed in the first 3 days after the admission. RESULTS: Among 42 enrolled patients, 19 (45%) had acute renal involvement (Group A) ; 23 (55%) (Group B) had normal DMSA scan (n = 16), or old scarring (n = 4) or various anomalies related to uropathy (n = 3). In group A, the mean PCT level was significantly higher than in the group B (5.4 ng/ml, vs 0.4 ng /ml, p < 10(-5)). In these 2 groups, mean C reactive protein (CRP) levels were 99.1 mg/l and 44.6 mg/l respectively (p < 0.001). For a level of serum PCT > or = 0.5 ng/ml, the sensitivity and specificity to predict the renal involvement were 100% and 87% respectively; for a level> or= 20 mg/l CRP had a sensitivity of 94% but a specificity of 30%. CONCLUSION: Serum PCT levels were significantly increased in febrile children with UTI when acute renal parenchymal involvement was present. PCT seems a better marker than CRP for the prediction of patients at risk of renal lesions. PMID- 15893463 TI - [Anthropometric parameters, frequency and risk factors of intrauterine growth retardation in full-term infants in North Togo]. AB - OBJECTIVE: To establish fullterm newborn anthropometric parameters to determine the frequency of intrauterine growth retardation and to study mother's and socio economic risk factors associated with the intrauterine growth retardation in the population in North-Togo. METHODS: Twelve month prospective study in the savana area in North-Togo. One thousand and six alive newborns born after normal single pregnancy, and without severe neonatal diseases were included. RESULTS: The mean birth-weight +/- SD was 2938 +/- 421 g. The incidence of low birth-weight was 11,7%. Among mother's and socio-economic risk factors, teenage mothers, mothers with a weight lower than 50 kg or with body mass index lower than 20 kg/m2 had a significantly high risk of having a child with intrauterine growth retardation (Odds ratio = 2,71; 2,57; 1,57 respectively). Father's job also affected the mother's and newborn's weight. CONCLUSION: The recognition of these easy to identify risk factors should allow specific recommendations for this population. PMID- 15893464 TI - Clinical practice guidelines: medical follow-up of patients with asthma--adults and adolescents. AB - The follow-up of patients with asthma should focus on asthma control (disease course over a number of weeks). PMID- 15893465 TI - Low incidence of paradoxical bronchoconstriction in asthma and COPD patients during chronic use of Respimat soft mist inhaler. AB - Respimat Soft Mist Inhaler (SMI) is a new-generation inhaler that offers improved lung deposition compared with chlorofluorocarbon metered dose inhalers (CFC MDIs). Bronchodilators administered via Respimat SMI are preserved and stabilised with low concentrations of benzalkonium chloride and ethylene diamine tetra acetic acid, both of which have been reported to cause dose-related paradoxical bronchoconstriction. The aim of this analysis was to compare the incidence of paradoxical bronchoconstriction after chronic use of bronchodilators via Respimat SMI and CFC-MDI. Data from three clinical trials, in which patients with asthma or chronic obstructive pulmonary disease (COPD) received ipratropium bromide alone or in combination with fenoterol hydrobromide, or placebo via Respimat SMI or CFC-MDI for 12 weeks, were included in the analysis. In order to evaluate the risk of paradoxical bronchoconstriction, we identified four respiratory events that might have occurred within 30 min of inhalation on four test days; these were: 'bronchospasm', 'other respiratory adverse events', 'rescue medication use' and 'asymptomatic drop in FEV(1) 15% from baseline'. In total, 631 asthma and 1538 COPD patients participated in the three studies. No occurrences of bronchospasm were reported with Respimat SMI on any test day. Overall, the incidence of respiratory events possibly indicative of paradoxical bronchoconstriction was low and similar for both devices. There was no increase in the incidence of events during 12 weeks' treatment. Delivery of bronchodilators by Respimat SMI is safe with regard to paradoxical bronchoconstriction during chronic use in patients with asthma or COPD. PMID- 15893466 TI - The biosensor based on the pyruvate oxidase modified conducting polymer for phosphate ions determinations. AB - An enzymatic biosensor was fabricated by the covalent immobilization of pyruvate oxidase (PyO) onto the nano-particle comprised poly-5,2':5',2''-terthiophene-3' carboxylic acid, poly-TTCA (nano-CP) layers on a glassy carbon electrode (GCE) for the amperometric detection of the phosphate ions. The direct electron transfer reaction of the immobilized PyO onto the nano-CP layers was investigated and the electron transfer rate constant was determined to be 0.65 s(-1). The electrochemically prepared nano-CP lowered the oxidation potential (+0.40 V versus Ag/AgCl) of an enzymatically generated H(2)O(2) by PyO in a phosphate solution. Experimental parameters affecting the sensitivity of the biosensors, such as amounts of the cofactors, the pH, the applied potential, and the temperature were optimized. A linear response for the detection of the phosphate ion was observed between 1.0 microM and 100 microM and the detection limit was determined to be about 0.3 microM. The response time of the biosensors was about 6s. The biosensor showed good selectivity towards other interfering anions. The long-term storage stability of the phosphate biosensor was studied and the sensor was applied in a human serum sample for the phosphate ions detection. PMID- 15893467 TI - 17beta-estradiol rescues spinal motoneurons from AMPA-induced toxicity: a role for glial cells. AB - The ability of astrocytes to mediate 17beta-estradiol neuroprotection of spinal motoneurons challenged with AMPA has been evaluated in a co-culture system in which pure motoneurons were pulsed with 20 microM AMPA and then transferred onto an astrocyte layer pretreated for 24 h with 10 nM 17beta-estradiol. Under these conditions, AMPA toxicity was reverted, an effect that was likely related to increased production and release of GDNF, as shown by RT-PCR, Western blot analysis and ELISA assay. In addition, treatment with GDNF during the 24 h that followed the AMPA pulse produced a similar neuroprotective effect, whereas addition of a neutralizing anti-GDNF antibody prevented neuroprotection. These data suggest a role for astrocytes in the neuroprotective effect of 17beta estradiol against spinal motoneuron death and find strong support in the marked up-regulation of estrogen receptor alpha found in spinal astrocytes of amyotrophic lateral sclerosis patients. PMID- 15893468 TI - Aggravation of ischemic brain injury by prion protein deficiency: role of ERK-1/ 2 and STAT-1. AB - The cellular isoform of prion protein, PrPc, may confer neuroprotection in the brain, according to recent studies. To elucidate the role of PrPc in stroke pathology, we subjected PrPc-knockout (Prnp(0/0)), wild-type and PrPc-transgenic (tga20) mice to 30 min of intraluminal middle cerebral artery occlusion, followed by 3, 24 or 72 h reperfusion, and examined how PrPc levels influence brain injury and cell signaling. In immunohistochemical experiments and Western blots, we show that PrPc expression is absent in the brains of Prnp(0/0) mice, detectable in wild-type controls and approximately 4.0-fold elevated in tga20 mice. We provide evidence that PrPc deficiency increases infarct size by approximately 200%, while transgenic PrPc restores tissue viability, albeit not above levels in wild-type animals. To elucidate the mechanisms underlying Prnp(0/0)-induced injury, we performed Western blots, which revealed increased activities of ERK-1/-2, STAT-1 and caspase-3 in ischemic brains of Prnp(0/0)mice. Our data suggest a role of cytosolic signaling pathways in Prnp(0/0)-induced cell death. PMID- 15893469 TI - Topical use of liposomal copper palmitate formulation blocks porphyrin-induced photosensitivity in rats. AB - Photodynamic therapy (PDT) is a new treatment modality that uses porphyrin derivatives and visible light, especially for the treatment of cancer. However, PDT with certain photosensitisers can cause prolonged skin photosensitization. This is particularly true for Photofrin II (Photofrin)-mediated PDT where patients are required to avoid direct exposure to sunlight for a period of 4-6 weeks. This is the only long-term adverse reaction to the drug. Recent studies have shown that topical copper treatment avoids this type of inflammatory reaction. In this study, we have tested the efficiency of the liposomal formulation of copper palmitate on porphyrin-photosensitized rats. Initially, adult male Sprague-Dawley rats were rendered photosensitive either by administration of Photofrin or aminolevulinic acid (ALA), a precursor of protoporphyrin IX (PpIX). Prior to this, their dorsal skin was shaved and treated topically with a cream consisting of either empty or copper palmitate encapsulated liposomal formulation. After being kept in a dimmed light environment, the rats were exposed to visible light, and inflammatory responses were inspected. Histological studies revealed that no inflammatory cells were present at the skin sites treated with liposomal cream containing copper palmitate in the Photofrin-sensitized group while no reduction in the number of inflammatory cells was observed at the skin samples treated with the empty liposomes. In conclusion, the data demonstrate the significant protective effect of topically-applied liposome-encapsulated copper palmitate against both Photofrin and ALA-induced PpIX photosensitivity. PMID- 15893470 TI - Photodegradation and photosensitization of mycosporine-like amino acids. AB - The photodegradation and photosensitization of several mycosporine-like amino acids (MAAs) were investigated. The photodegradation of the MAA, palythine, was tested with three photosensitizers: riboflavin, rose bengal and natural seawater. For comparison of degradation rates, the riboflavin-mediated photosensitization of six other MAAs was also examined. When riboflavin was used as a photosensitizer in distilled water, MAAs were undetectable after 1.5h. Palythine showed little photodegradation when rose bengal was added as the photosensitizer (k=0.12x10(-3)m(2)kJ(-1)). Palythine dissolved in natural seawater containing high nitrate concentrations also showed slow photodegradation rate constants (k=0.26x10(-3)m(2)kJ(-1)) over a 24-h period of constant irradiation. Similar experiments in deep seawater with porphyra-334 and shinorine resulted in 75% of the initial MAA remaining after 4h of irradiation and rates of 0.018 and 0.026x10(-3) m(2) kJ(-1), respectively. Experiments conducted in deep seawater with riboflavin additions resulted in photodegradation rate constants between 0.77x10(-3) and 1.19x10(-3)m(2)kJ(-1) for shinorine and porphyra-334, respectively. Photoproduct formation appeared to be minimal with the presence of a dehydration product of the cycloheximine ring structure indicated as well as the presence of amino acids. Evidence continues to build for the role of MAAs as potent and stable UV absorbers. This study further highlights the photostability of several MAAs in both distilled and seawater in the presence of photosensitizers. PMID- 15893471 TI - Is arginine a protein-denaturant? AB - Arginine is a useful solvent additive for many applications, including refolding and solubilization of proteins from insoluble pellets, and suppression of protein aggregation and non-specific adsorption during formulation and purification. However, there is a concern that arginine may be a protein-denaturant, which may limit the expansion of its applications. Such concern arises from the facts that arginine decreases melting temperature and perturbs the spectroscopic properties of certain proteins and contains a guanidinium group, which is a critical chemical structure for denaturing activity of guanidine hydrochloride. Here, we show that although arginine does lower the melting temperatures of certain proteins, the extent is insufficient to cause denaturation of proteins at or below room temperature. The proteins described here show enzymatic activity and folded structure in the presence of arginine, although the local structure around aromatic amino acids is perturbed by arginine. Arginine differs from guandinine hydrochloride in the mode of interactions with proteins, which may be a primary reason why arginine is not a protein-denaturant. PMID- 15893472 TI - Development and function of the thymus in teleosts. AB - The thymus plays a pivotal role in the development of the adaptive immune system, an important factor that separates higher vertebrates from the rest of the animal phyla. The development of functional T-cells from thymocytes is a crucial step in the development of a functional vertebrate immune system and whilst recent advances in molecular and developmental biology have advanced our understanding of T-cell development, they have also provided potential model species across the vertebrate phyla including the zebrafish (Danio rerio). However, this species is one of more than 20,000 species of fish that could assist in elucidating the development of the vertebrate thymus and, consequently, the evolution of the vertebrate immune response. In this paper we review the knowledge of the teleost thymus through the organogenesis and development studies in teleosts together with advances in molecular and functional approaches. Where necessary we will combine this knowledge with that obtained in higher vertebrates. PMID- 15893473 TI - Language network specializations: an analysis with parallel task designs and functional magnetic resonance imaging. AB - Although the classical core regions of the language system (Broca's and Wernicke's areas) were defined over a century ago, it took the advent of functional imaging to sharpen our understanding of how these regions and adjacent parts of the brain are associated with particular aspects of language. One limitation of such studies has been the need to compare results across different subject groups, each performing a different type of language task. Thus, this study was designed to examine overlapping versus segregated brain activations associated with three fundamental language tasks, orthography, phonology and semantics performed by the same subjects during a single experimental session. The results demonstrate a set of primarily left-sided core language regions in ventrolateral frontal, supplementary motor, posterior mid-temporal, occipito temporal and inferior parietal areas, which were activated for all language tasks. Segregated task-specific activations were demonstrated within the ventrolateral frontal, mid-temporal and inferior parietal areas. Within the inferior frontal cortex (Broca's regional complex), segregated activations were seen for the semantic and phonological tasks. These findings demonstrate both common and task specific activations within the language system. PMID- 15893474 TI - Scanning silence: mental imagery of complex sounds. AB - In this functional magnetic resonance imaging (fMRI) study, we investigated the neural basis of mental auditory imagery of familiar complex sounds that did not contain language or music. In the first condition (perception), the subjects watched familiar scenes and listened to the corresponding sounds that were presented simultaneously. In the second condition (imagery), the same scenes were presented silently and the subjects had to mentally imagine the appropriate sounds. During the third condition (control), the participants watched a scrambled version of the scenes without sound. To overcome the disadvantages of the stray acoustic scanner noise in auditory fMRI experiments, we applied sparse temporal sampling technique with five functional clusters that were acquired at the end of each movie presentation. Compared to the control condition, we found bilateral activations in the primary and secondary auditory cortices (including Heschl's gyrus and planum temporale) during perception of complex sounds. In contrast, the imagery condition elicited bilateral hemodynamic responses only in the secondary auditory cortex (including the planum temporale). No significant activity was observed in the primary auditory cortex. The results show that imagery and perception of complex sounds that do not contain language or music rely on overlapping neural correlates of the secondary but not primary auditory cortex. PMID- 15893475 TI - Estimating the global order of the fMRI noise model. AB - One of the major issues in GLM-based fMRI analysis techniques is the presence of temporal autocorrelations in the residual signal after regression. A possible correction method is that of prewhitening, which fits an autoregressive (or other) model to the residual and uses the expected temporal autocorrelations of the model to transform the data and design matrix such that the residual becomes white noise. In this article, a method is introduced to estimate the global autoregressive model order of a data set, based on the residuals after regression. The proposed global standardized partial autocorrelation (SPAC) method tests whether the spatial profile of partial autocorrelations at a certain lag is random, and uses random field theory to account for the spatial correlations typical for fMRI data. It is tested both on synthetic and fMRI data, and is compared to two traditional techniques for model order estimation. PMID- 15893476 TI - Volumetric vs. surface-based alignment for localization of auditory cortex activation. AB - The high degree of intersubject structural variability in the human brain is an obstacle in combining data across subjects in functional neuroimaging experiments. A common method for aligning individual data is normalization into standard 3D stereotaxic space. Since the inherent geometry of the cortex is that of a 2D sheet, higher precision can potentially be achieved if the intersubject alignment is based on landmarks in this 2D space. To examine the potential advantage of surface-based alignment for localization of auditory cortex activation, and to obtain high-resolution maps of areas activated by speech sounds, fMRI data were analyzed from the left hemisphere of subjects tested with phoneme and tone discrimination tasks. We compared Talairach stereotaxic normalization with two surface-based methods: Landmark Based Warping, in which landmarks in the auditory cortex were chosen manually, and Automated Spherical Warping, in which hemispheres were aligned automatically based on spherical representations of individual and average brains. Examination of group maps generated with these alignment methods revealed superiority of the surface-based alignment in providing precise localization of functional foci and in avoiding mis-registration due to intersubject anatomical variability. Human left hemisphere cortical areas engaged in complex auditory perception appear to lie on the superior temporal gyrus, the dorsal bank of the superior temporal sulcus, and the lateral third of Heschl's gyrus. PMID- 15893477 TI - A Bayesian approach to the estimation of ancestral genome arrangements. AB - We describe a Bayesian approach to estimate phylogeny and ancestral genome arrangements on the basis of genome arrangement data using a model in which gene inversion is the sole mechanism of change. While we have described a similar method to estimate phylogenetic relationships in the statistics literature, the novel contribution of the present work is the description of a method to compute probability distributions of ancestral genome arrangements. We assess the robustness of posterior distributions to different specifications of prior distributions and provide an empirical means to selecting a prior distribution. We note that parsimony approaches to ancestral reconstruction in the literature focus on the development of computationally efficient algorithms for searching for optimal ancestral genome arrangements, but, unlike Bayesian approaches, do not include assessment of uncertainty in these estimates. We compare and contrast a Bayesian approach with a parsimony approach to infer phylogenies and ancestral arrangements from genome arrangement data by re-analyzing a number of previously published data sets. PMID- 15893478 TI - Seizure-related factors and non-verbal intelligence in children with epilepsy. A population-based study from Western Norway. AB - PURPOSE: To study the relationship between seizure-related factors, non-verbal intelligence, and socio-economic status (SES) in a population-based sample of children with epilepsy. METHODS: The latest ILAE International classifications of epileptic seizures and syndromes were used to classify seizure types and epileptic syndromes in all 6-12 year old children (N=198) with epilepsy in Hordaland County, Norway. The children had neuropediatric and EEG examinations. Of the 198 patients, demographic characteristics were collected on 183 who participated in psychological studies including Raven matrices. 126 healthy controls underwent the same testing. Severe non-verbal problems (SNVP) were defined as a Raven score at or <10th percentile. RESULTS: Children with epilepsy were highly over-represented in the lowest Raven percentile group, whereas controls were highly over-represented in the higher percentile groups. SNVP were present in 43% of children with epilepsy and 3% of controls. These problems were especially common in children with remote symptomatic epilepsy aetiology, undetermined epilepsy syndromes, myoclonic seizures, early seizure debut, high seizure frequency and in children with polytherapy. Seizure-related characteristics that were not usually associated with SNVP were idiopathic epilepsies, localization related (LR) cryptogenic epilepsies, absence and simple partial seizures, and a late debut of epilepsy. Adjusting for socio-economic status factors did not significantly change results. CONCLUSIONS: In childhood epilepsy various seizure-related factors, but not SES factors, were associated with the presence or absence of SNVP. Such deficits may be especially common in children with remote symptomatic epilepsy aetiology and in complex and therapy resistant epilepsies. Low frequencies of SNVP may be found in children with idiopathic and LR cryptogenic epilepsy syndromes, simple partial or absence seizures and a late epilepsy debut. Our study contributes to an overall picture of cognitive function and its relation to central seizure characteristics in a childhood epilepsy population and can be useful for the follow-up team in developing therapy strategies that meet the individual needs of the child with epilepsy. PMID- 15893479 TI - Suppression of macrophage infiltration into the conjunctiva by clodronate liposomes in experimental immune-mediated blepharoconjunctivitis. AB - Macrophages infiltrate the conjunctiva in severe cases of allergic conjunctivitis (AC) such as atopic keratoconjunctivitis (AKC). We established experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats as a model for severe types of AC. We investigated whether macrophage infiltration in the conjunctiva in this EC model is inhibited by clodronate liposomes (CL2MDP-lip). The numbers of ED1-positive but not ED2-positive macrophages in the conjunctivas were increased by the induction of EC. Subconjunctival injection of CL2MDP-lip decreased the number of ED2-positive but not ED1-positive macrophages in the conjunctivas of naive rats. CL2MDP-lip did not affect macrophages in the spleen. Subconjunctival injection of CL2MDP-lip into EC-developing BN rats decreased the number of ED2-positive macrophages at all the time points. ED1-positive cell infiltration was inhibited when treatment was administered just prior to OVA challenge. Intravenous injection of CL2MDP-lip decreased the number of ED2 positive cells in the conjunctiva. Thus, we conclude that CL2MDP-lip inhibits infiltration of macrophages into the conjunctiva within 24 h of antigen challenge. PMID- 15893480 TI - Intracellular localization of Xenopus small heat shock protein, hsp30, in A6 kidney epithelial cells. AB - Small heat shock proteins (shsps) are molecular chaperones that are inducible by environmental stress. In this study, immunocytochemical analysis and laser scanning confocal microscopy revealed that the shsp family, hsp30, was localized primarily in the cytoplasm of Xenopus A6 kidney epithelial cells after heat shock or sodium arsenite treatment. Heat shock-induced hsp30 was enriched in the perinuclear region with some immunostaining in the nucleus but not in the nucleolus. In sodium arsenite-treated cells hsp30 was enriched towards the cytoplasmic periphery as well as showing some immunostaining in the nucleus. At higher heat shock temperatures (35 degrees C) or after 10 microM sodium arsenite treatment, the actin cytoskeleton displayed some disorganization that co localized with areas of hsp30 enrichment. Treatment of A6 cells with 50 microM sodium arsenite induced a collapse of the cytoskeleton around the nucleus. These results coupled with previous studies suggest that stress-inducible hsp30 acts as a molecular chaperone primarily in the cytoplasm and may interact with cytoskeletal proteins. PMID- 15893481 TI - Catalases protect cellular proteins from oxidative modification in Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae cells had higher antioxidant enzyme activities under growth in ethanol than that in glucose as a carbon and energy source. The correlations between catalase activity and protein carbonyl level (r(2)=0.857), between catalase and glucose-6-phosphate dehydrogenase activities (r(2)=0.924) and between protein carbonyl levels and glucose-6-phosphate dehydrogenase activity (r(2)=0.988) under growth in ethanol were found. Growing in ethanol the strain deficient in cytosolic and peroxisomal catalases had 7.1-fold higher level of carbonyl proteins than that of wild-type strain. Our data suggest that in vivo catalases may protect glucose-6-phosphate dehydrogenase against oxidative inactivation. PMID- 15893482 TI - Hormonal interactions in Tetrahymena: effect of hormones on levels of epidermal growth factor (EGF). AB - Tetrahymena pyriformiswas treated with insulin, histamine or serotonin for 30 min and epidermal growth factor (EGF) level was studied inside the cells using specific antibodies and flow cytometry as well as confocal microscopy. The EGF concentration was highly significantly elevated after hormone treatment, regardless of the hormone used. EGF was localized mainly in the cortical region (mucocysts) and in vesicles and this localization did not differ in untreated and treated cells. The results call attention to the possibility of interactions between hormones at unicellular level and points to the presence of a hormonal system in Tetrahymena that includes receptors, hormones and signal transduction pathways as well as hormonal interactions. This could be the basis of further evolution to the hormonal system of multicellulars. PMID- 15893483 TI - Role of reactive oxygen species (ROS), metalloproteinase-2 (MMP-2) and interleukin-6 (IL-6) in direct interactions between tumour cell spheroids and endothelial cell monolayer. AB - Metastasis is a multistep process involving a variety of direct cell-cell, cell matrix and paracrine interactions. In the present study, we examined some consequences of direct interaction between tumour cells and endothelial cells in vitro. When multicellular spheroids of two human tumour cell lines (HeLa and Hep 2) were transferred onto a human umbilical vein endothelial cell (HUVEC) monolayer, a peri-spheroidal zone of damaged endothelial cells was observed after 24h co-culture. To determine the cause of this damage, the production levels of superoxide anion (O2-), interleukin-6 (IL-6) and metalloproteinase-2 (MMP-2) were measured both in co-culture and in monocultures of the tumour cell spheroids and endothelial cells. Attachment of HeLa and Hep-2 cellular spheroids to the HUVEC monolayer resulted in 1.6-fold and 2.1-fold increases in O2- release, respectively. Also, the MMP-2 level was five times greater in the co-culture than in the tumour spheroid monoculture. The increase of IL-6 in the co-culture model, on the other hand, was only slight. However, a 2h preincubation of endothelial cells with LPS (10 microg/ml) prior to the transfer of spheroids induced a significant increase in the production of this cytokine compared to an appropriate control (an LPS-activated endothelial cell monolayer). These results strongly suggest that both ROS and MMP-2 are involved in endothelial cell injury when tumour cells cross the endothelial barrier. Moreover, IL-6, which participates in the inflammatory response, may also be involved in the extravasation of tumour cells. PMID- 15893484 TI - A review of biological factors implicated in abdominal aortic aneurysm rupture. AB - Abdominal aortic aneurysm (AAA) rupture is the 13th commonest cause of death in the Western World. Although considerable research has been applied to the aetiology and mechanism of aneurysm expansion, little is known about the mechanism of rupture. Aneurysm rupture was historically considered to be a simple physical process that occurred when the aortic wall could no longer contain the haemodynamic stress of the circulation. However, AAAs do not conform to the law of Laplace and there is growing evidence that aneurysm rupture involves a complex series of biological changes in the aortic wall. This paper reviews the available data on patient variables associated with aneurysm rupture and presents the evidence implicating biological factors in AAA rupture. PMID- 15893485 TI - Non-penetrating clips for vascular anastomosis. PMID- 15893486 TI - Phenotypic and functional characterization of intestinal epithelial exosomes. AB - Intestinal epithelial cells (IEC) are located at a strategic position between the external environment and the most extended lymphoid tissue in the body. Besides their central role in the absorption of nutrients, IEC also provide antigenic information to the immune system and are involved in the balance tolerance/allergy to food antigens. Like professional antigen presenting cells, IEC have been shown to secrete 30- to 90-nm diameter vesicles named exosomes, in a polarized way, either from their apical or basolateral side. These vesicles carry molecules involved in adhesion and antigen presentation, comprising major histocompatibility complex (MHC) class I and class II molecules, tetraspan proteins, CD26/dipeptidyl-peptidase IV, and A33 antigen, a molecule essentially restricted to the intestinal epithelium. Invariant chain, transferrin receptor, and Na-K-ATPase are not expressed on epithelial exosomes. In vivo, in mice, epithelial exosomes carrying MHC/ovalbumin peptide complexes induce specific immune responses when injected intraperitoneally. A33 antigen, an Ig-like molecule highly specific for intestinal epithelial cells and enriched in epithelial exosomes, is found at the surface of cells entering mesenteric lymph nodes suggesting exosome migration from the epithelial layer to the gut associated lymphoid system. Taken together, intestinal epithelial exosomes released at the basolateral surface of enterocytes could be antigen-carrying structures constituting a link between luminal antigens and the local immune system and acting as sensors of the antigenic information present in the intestinal lumen. PMID- 15893487 TI - Optimization of the irradiation power in chemical exchange dependent saturation transfer experiments. AB - In chemical exchange dependent saturation transfer imaging experiments, exchangeable solute protons are saturated and the transfer of saturation to water is subsequently detected. When the applied irradiation power is comparable to the resonance frequency difference between the water protons and saturated solute protons, the proton transfer (PT) efficiency is reduced due to concomitant direct saturation effects. In this study, the PT process is modeled using a two-pool system. An empirical general proton transfer ratio (PTR) equation for arbitrary RF irradiation power is derived, and its optimal power to maximize the PTR is analyzed. The results are confirmed experimentally on 4.7 T using a poly-L-lysine solution. The theory provides a useful tool for optimizing the irradiation power of the PT sequences in the presence of direct saturation effects. PMID- 15893488 TI - Behavioural, ventilatory and respiratory responses of epigean and hypogean crustaceans to different temperatures. AB - Impact of temperature (from -2 to 28 degrees C) on survival, oxygen consumption, locomotory and ventilatory activities was measured in two aquatic subterranean crustaceans (Niphargus rhenorhodanensis and Niphargus virei) and in a morphologically close surface-dwelling crustacean (Gammarus fossarum). The hypogean N. virei presented all characteristics of a stenothermal organism: it showed small thermal plasticity and optimised its performance on a narrow range of temperature. In contrast, the epigean G. fossarum and more surprisingly the hypogean N. rhenorodanensis can be both characterized as eurythermal organisms: they exhibited important survival times and conserved their performance optimum throughout a large range of temperature. Such differences of survival and performance patterns in two hypogean organisms were unexpected since they both live in very thermally buffered biotopes. Our data suggest fresh hypotheses about the role of glaciations in the history and adaptation of hypogean crustaceans. PMID- 15893489 TI - Lipid composition of deep-sea hydrothermal vent tubeworm Riftia pachyptila, crabs Munidopsis subsquamosa and Bythograea thermydron, mussels Bathymodiolus sp. and limpets Lepetodrilus spp. AB - Lipid composition was determined for hydrothermal vent species collected by the Deep Submergence Vehicle ALVIN from chimneys at 2,500 m depth on the East Pacific Rise. These are the first lipid biomarker studies for most of these species. Lipid content was low and dominated by polar lipid in the vestimentiferan tubeworm Riftia pachyptila, mussels Bathymodiolus sp. and limpets Lepetodrilus spp. The galatheid (Munidopsis subsquamosa) and most brachyuran adult (Bythograea thermydron) crabs were characterized by higher storage lipid (triacylglycerol). Total polyunsaturated fatty acids were similar in R. pachyptila plume and body, but higher in the posterior part of the soft body, which had more docosahexaenoic acid (2-5% of total FA) compared to the anterior and plume (< or =0.3%). Two sulphur-oxidizing bacterial markers, 16:1(n-7)c and 18:1(n-7)c, were high in R. pachyptila and mussel (up to 23%), but lower in both crab species (4-17%). R. pachyptila had greater nonmethylene interrupted diunsaturated fatty acids (8-13%) than all other species (2-8%). R. pachyptila may desaturate and elongate 18:1(n 7)c to obtain essential polyunsaturated fatty acids 20:5(n-3) and 20:4(n-6). The sterol composition of R. pachyptila included similar amounts of cholesterol and desmosterol, whereas the other species had a more diverse sterol composition. These differences in lipids, fatty acids and sterols reflect diverse nutritional strategies and possibly temperature regimes in these species. PMID- 15893490 TI - [Hope and limits in cell therapies]. AB - The hematopoietic system is one of the best characterized human cellular system in which a multipotent adult stem cell is at the origin of all the cells of a tissue. These last years, the use of stem cells has given rise to many hopes in regenerative medicine, especially in diseases without efficient therapies. However the hematopoietic system in which stem cell transplantation has entered in clinical practice for many years has also shown the limits of these approaches. Especially the in vitro manipulation of hematopoietic stem cells remains a challenge which requires more fundamental knowledge on the biology of stem cells including self renewal and homing. Knowledge for other tissue system is even more preliminary but the same experimental strategy used for hematopoietic stem cell can be translated and may accelerate their use in cell therapies. Characterization of human adult pluripotent stem cells and the generation of human ES cells capable to differentiate towards several tissues have led to new hopes but the road to their use in therapies may be long and will require a lot of investment in basic biology. PMID- 15893491 TI - Establishment of a biological assay system for human retroviral protease activity. AB - In order to obtain indicator cell lines that are exquisitely susceptible to human T-lymphotropic virus type 1 (HTLV-1), luciferase gene driven by HTLV-1 long terminal repeat (LTR) was transfected into lymphocytic H9 cells with neo gene, and cell lines were selected by G418. A cell line (H9/K30luc) was found to produce an extremely high level of luciferase only when co-cultured with HTLV-1 producer MT-2 cells. Both in the absence and presence of a reverse transcriptase (RT) inhibitor azidothymidine, H9/K30luc cells generated similarly high luciferase activity upon co-cultivation with MT-2 cells. To develop an equivalent system for human immunodeficiency virus type 1 (HIV-1), H9/NL432 cells, which are stably infected with HIV-1 and producing a low level of the virus-like MT-2 cells for HTLV-1, were generated. Together with the indicator cell line H9/H1luc for HIV-1 already reported, antiviral effects of some agents on HTLV-1 and HIV-1 could be readily and sensitively evaluated by similar methods. In fact, by using our system, an HIV-1 protease inhibitor, saquinavir, was demonstrated to be highly effective against HIV-1 but not against HTLV-1. PMID- 15893492 TI - Inter-species genetic movement may blur the epidemiology of streptococcal diseases in endemic regions. AB - Streptococcus dysgalactiae subsp. equisimilis (human group G streptococcus, GGS) is generally regarded as a commensal organism but can cause a spectrum of human diseases very similar to that caused by S. pyogenes (group A streptococcus, GAS). Lateral acquisition of genes between these two phylogenetically closely related species is well documented. However, the extent and mechanisms of lateral acquisitions is not known. We report here genomic subtraction between a pathogenic GGS isolate and a community GGS isolate and analyses of the gene sequences unique to the pathovar. Our results show that cross-species genetic transfers are common between GGS and two closely related human pathogens, GAS and the group B streptococcus. We also demonstrate that mobile genetic elements, such as phages and transposons, play an important role in the ongoing inter-species transfers of genetic traits between extant organisms in the community. Furthermore, lateral gene transfers between GAS and GGS may occur more frequently in geographical regions of high GAS endemicity. These observations may have important implications in understanding the epidemiology of streptococcal diseases in such regions. PMID- 15893493 TI - CD4+CD25+ regulatory T cells in HIV infection. AB - The immune system faces the difficult task of discerning between foreign, potentially pathogen-derived antigens and self-antigens. Several mechanisms, including deletion of self-reactive T cells in the thymus, have been shown to contribute to the acceptance of self-antigens and the reciprocal reactivity to foreign antigens. Over the last decade it has become increasingly clear that CD4(+)CD25(+) T(Reg) cells are crucial for maintenance of T cell tolerance to self-antigens in the periphery, and to avoid development of autoimmune disorders. Recently, evidence has also emerged that demonstrates that CD4(+)CD25(+) T(Reg) cells can also suppress T cell responses to foreign pathogens, including viruses such as HIV. In this article we review the current knowledge and potential role of CD4(+)CD25(+) T(Reg) cells in HIV infection. PMID- 15893494 TI - Human CD4+ regulatory T cells and activation-induced tolerance. AB - In the past years growing attention has been given to the role of regulatory T (Tr) cells in inducing and monitoring peripheral tolerance. Various subsets of Tr cells have been described based on their surface phenotype and cytokine production. However, presently there are no specific reliable markers for any of the Tr subsets and their classification is based predominantly upon their mode of suppression. In addition, the molecular mechanisms underlying the induction and mode of action of all Tr cell subsets remain to be elucidated. Here we review recent developments regarding human CD4+ Tr cells, their origin, phenotype, antigen specificity and mode of suppression. PMID- 15893495 TI - Identification of a detrimental role for NK cells in pneumococcal pneumonia and sepsis in immunocompromised hosts. AB - Gram-positive sepsis is a major disease problem. However, the contribution of various immune cell types to pathogenesis remains unclear. By infecting scid and wild type BALB/c mice with Streptococcus pneumoniae we have found a situation in which natural killer (NK) cells can play a detrimental role in the response to infection. scid mice were found to be significantly more susceptible to local and systemic pneumococcal infection than controls; they had significantly higher bacterial loads, elevated inflammatory responses and more widespread lung pathology. Interestingly, depletion of NK cells in scid mice resulted in significantly lower bacteraemia and inflammatory cytokine production. Infection with pneumococci deficient in pneumolysin revealed the toxin was involved in cytokine production. Overall results indicate that elevated NK cell activity during pneumococcal pneumonia amplifies pulmonary and systemic inflammation, increases bacteraemia and results in poor outcome. PMID- 15893496 TI - Microbial compounds induce the expression of pro-inflammatory cytokines, chemokines and human beta-defensin-2 in vaginal epithelial cells. AB - Vaginal epithelium has a powerful innate immune system that protects the female reproductive organs from bacterial and fungal infections. In the present study, we aimed to explore whether the Toll-like receptor (TLR) signaling pathway and the induction of pro-inflammatory cytokines and antimicrobial peptides could contribute to the protection against pathogenic microorganisms in vaginal epithelia, using an immortalized vaginal epithelial cell line PK E6/E7 as a model. We found that TLR2 and TLR4 receptors are expressed in vivo in the vaginal epithelia and in vitro in PK E6/E7 vaginal epithelial cell line. The Gram negative cell wall compound lipopolysaccharide (LPS), the Gram-positive compound peptidoglycan (PGN), heat-killed Candida albicans and zymosan significantly (P<0.05) induced the expression of pro-inflammatory cytokines and chemokines such as TNF-alpha and IL-8/CXCL8 in vaginal epithelial cells. Furthermore, the expression and production of human beta-defensin-2 (hBD2), an antimicrobial peptide with chemotactic functions, was also up-regulated in PK E6/E7 cells after treatment with LPS, PGN or C. albicans. Treatment of vaginal epithelial cells with microbial compounds induced the activation and nuclear translocation of NF kappaB transcription factor, a key element of innate and adaptive immune responses. In our work, we provide evidence that microbial compounds induce the production of pro-inflammatory cytokines, chemokines and antimicrobial peptides in vaginal epithelial cells. In vivo, vaginal epithelial cell-derived inflammatory mediators and antimicrobial peptides may play important roles in vaginal immune responses and in the elimination of pathogens from the female reproductive tract. PMID- 15893497 TI - Vaccinia virus infection and gene transduction in cultured neurons. AB - The study of neurons in culture would benefit from the development of a gene transduction system capable of delivering foreign genes at high efficiency, as transduction of primary neurons with existing systems is inefficient. The efficacy of lytic vaccinia virus (VV) infection of primary retinal cultures and PC12 cells (a model of neuronal differentiation) was examined in order to determine the efficiency of gene transduction using VV in neuronal primary culture. VV was able to infect retinal cells and PC12 cells and express transgenes of Escherichia coli beta-galactosidase (lacZ) and epithelial fatty acid binding protein (E-FABP) in a virus dose-dependent manner. Most (50-100%) of the retinal cells were positive for transgene protein at multiplicities of infection (MOI) between 10 and 100 plaque-forming units (PFU), while over 50% of VV-infected PC12 cells expressed the virus encoded gene at an MOI = 10. The production of foreign mRNA and protein by VV following infection was verified by PCR and Western blot. Because VV is a lytic virus, cytopathic effects were examined. Retinal cultures maintained for 0.5 days in vitro showed greater than 90% survival at 24 h post-infection, while 14-day cultures were equally viable for 48 h. Retinal ganglion cells and differentiated PC12 cells appear to be more protected against lytic VV infection than proliferating glial and undifferentiated PC12 cells. These data suggest that VV may be a useful vector for delivering foreign genes to neuronal cells with an efficient transient transgene expression. PMID- 15893498 TI - Adoptive transfer of NK 1.1+ lymphocytes in immune-mediated colitis: a pro inflammatory or a tolerizing subgroup of cells? AB - T lymphocytes expressing NK1.1 marker (NKT) have been suggested to play crucial roles in immune modulation. AIM: To determine the role of NK1.1+ cells in induction and maintenance of pro-inflammatory and/or tolerizing responses. METHODS: Colitis was induced in C57/B6 donor mice by intracolonic instillation of trinitrobenzenesulfonic acid (TNBS). Donor mice received five oral doses of colonic proteins extracted from TNBS-colitis colonic wall. Depletion of NK1.1+ lymphocytes was performed before lymphocyte harvesting. Splenocytes were harvested and separated into T-cell subpopulations, and transplanted into recipient mice before intracolonic instillation of TNBS. Standard clinical, macroscopic, and microscopic scores, and intracellular staining, flow cytometry, and cytotoxicity assays were performed. RESULTS: The adoptive transfer of CD4+ and NK1.1+ cells harvested from tolerized mice markedly ameliorated the colitis in recipient mice. In contrast, the adoptive transfer of CD8+ and double negative lymphocytes failed to transfer the tolerance. Recipients of splenocytes from tolerized mice exhibited an increase in CD4+ IL4+/CD4+ IFNgamma+ ratio. In contrast, recipients of splenocytes from NK1.1-depleted-tolerized mice exhibited severe colitis with a significant decrease of the CD4+ IL4+/CD4+ IFNgamma+ ratio. However adoptive transfer of splenocytes from non-tolerized NKT-depleted mice led to an alleviation of colitis with a relative increase of the CD4+ IL4+/CD4+ IFNgamma+ ratio. CONCLUSIONS: NK1.1+ lymphocytes play a critical role in immune regulation. They may be accountable for an alteration of the inflammatory response and the CD4+ IL4+/CD4+ IFNgamma ratio immune-mediated colitis and in peripheral tolerance induction. PMID- 15893499 TI - [Accuracy of MR imaging combined with sonography for the diagnosis of persistent adnexal masses during pregnancy: about nine cases]. AB - OBJECTIVE: To evaluate the accuracy of MR imaging combined with sonography for the diagnosis of persistent adnexal masses during pregnancy. PATIENTS AND METHODS: From January 1999 to November 2003, nine patients with a persistent adnexal masse were included in this retrospective study. All patients underwent both transvaginal and transabdominal sonography combined with MR imaging. Accuracy of imaging techniques was evaluated by comparison with histology. RESULTS: Adnexal masse was detected by systematic sonography in six women whereas three patients were symptomatic. Mean gestational age at the diagnosis was 17 weeks of amenorrhea (range 5-36). Mean tumor size was 96 mm (range 2-15). Two patients underwent a surgery during pregnancy; one for acute abdominal pain related to an adnexal torsion and the second for suspicion of malignancy. One patient underwent a caesarean delivery for obstetrical reasons. In all patients, benign ovarian tumours were found. For eight patients the diagnosis suspected by imaging techniques was confirmed by histology. DISCUSSION AND CONCLUSION: The combination of sonography and MR imaging allows diagnosing accurately adnexal masses during pregnancy. PMID- 15893501 TI - Body mass index in Parkinson's disease. PMID- 15893502 TI - Developing terminology for documenting perioperative nursing interventions. AB - OBJECTIVE: To develop terminology for documenting perioperative nursing interventions. METHOD: Nursing documentation of 250 patients was explored by content analysis. The study was conducted to validate further the Perioperative Nursing Data Set (PNDS) in Finland. The data included 3442 terms that were clustered into 104 nursing interventions. A Delphi-panel (N=5) evaluated the relevance of each term on a scale from 1 to 4. The structure of each nursing intervention was checked to meet the minimum requirements of the ISO reference terminology model for nursing action concepts. Finally, the nursing interventions were compared with the original PNDS-interventions. RESULTS: The Delphi-panel accepted 98 nursing interventions with Content Validity Index from 0.88 to 1.0. Of the 133 nursing interventions in the PNDS, 60% (n=80) were covered by nursing interventions identified in this study. A notable portion of the nursing interventions related to preventive aspects of perioperative care and the patients' coping with perioperative experience. CONCLUSION: The findings supported the need for cross-cultural validation of a nursing language prior to clinical implementation. The ISO reference terminology model is recommended as framework in constructing concepts for nursing interventions in a regular form. PMID- 15893503 TI - The effect of environmental stress on absolute and mass-specific scope for growth in Daphnia magna Strauss. AB - Daphnids were reared for 2 weeks in different concentrations of food or cadmium, and growth and reproduction were measured as endpoints. At the end of the 14-day experimental period, scope for growth (SFG) was measured and expressed per individual (mJ/ind/h=absolute SFG) and per mg dry weight (mJ/mg/h=mass-specific SFG). Both food deprivation and cadmium stress decreased body size, and absolute SFG decreased with decreasing body size in both exposure scenarios. Also mass specific SFG decreased with decreasing body size under cadmium exposure, but an increase in mass-specific SFG was observed in the food ration experiment. This suggested that cadmium stress, apart from decreasing energy assimilation, also disturbs energy metabolism. Changes in both absolute and mass-specific SFG were mainly determined by changes in energy uptake, whereas energy loss varied little in response to both environmental stressors. With the cadmium-stressed daphnids, reproduction correlated positively with both absolute and mass-specific SFG. With the food-stressed daphnids however, reproduction correlated positively with absolute SFG but negatively with mass-specific SFG. Mass-corrected SFG still decreased with increasing cadmium stress, but did not differ between ration groups. Thus, mass-corrected SFG provides an indication of metabolic functioning, but appears less suited as an indicator of reproduction. PMID- 15893504 TI - Modulation of hypoglossal motoneuron excitability by intracellular signal transduction cascades. AB - Motoneuronal excitability is highly modulated by various inputs; however, comparatively little is known about postsynaptic signal transduction cascades that affect motoneuron excitability. In this review, we discuss the role of intracellular signaling cascades in the modulation of respiratory motoneuronal excitability. In particular, protein kinases and phosphatases dynamically and constitutively modulate respiratory-modulated inputs to XII motoneurons: (i) activation of protein kinase A (PKA) potentiates both excitatory and inhibitory drive currents; (ii) protein kinase G (PKG) depresses excitatory currents, and (iii) inhibition of protein phosphatases potentiates excitatory drive currents. We also describe a novel form of persistent plasticity (in vitro long-term facilitation; ivLTF) of motoneuronal output. ivLTF is induced by episodic activation of 5-HT(2) or alpha(1)-adrenoreceptors and is manifested as an increase in the amplitude of XII nerve output due to an increase in alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-mediated motoneuronal drive currents. Blockade of Group 1 metabotropic glutamate receptors or protein kinase C (PKC) prevents the induction of ivLTF. PMID- 15893505 TI - Protein-water displacement distributions. AB - The statistical properties of fast protein-water motions are analyzed by dynamic neutron scattering experiments. Using isotopic exchange, one probes either protein or water hydrogen displacements. A moment analysis of the scattering function in the time domain yields model-independent information such as time resolved mean square displacements and the Gauss-deviation. From the moments, one can reconstruct the displacement distribution. Hydration water displays two dynamical components, related to librational motions and anomalous diffusion along the protein surface. Rotational transitions of side chains, in particular of methyl groups, persist in the dehydrated and in the solvent-vitrified protein structure. The interaction with water induces further continuous protein motions on a small scale. Water acts as a plasticizer of displacements, which couple to functional processes such as open-closed transitions and ligand exchange. PMID- 15893506 TI - Disulfide-linked dimers of human adrenaline synthesizing enzyme PNMT are catalytically active. AB - The crystal structure of human phenylethanolamine N-methyltransferase (hPNMT) reveals a disulfide-linked dimer, despite the presence of reducing agent in the crystallisation conditions. By removing the reducing agent, hPNMT crystals grow more rapidly and at lower protein concentrations. However, it was unclear whether the disulfide bonds are only present in the crystal form or whether these affect enzyme activity. The solution oligomeric state of hPNMT was investigated using biochemical techniques and activity assays. We found that in the absence of reducing agent, hPNMT forms dimers in solution. Furthermore, the solution dimer of hPNMT incorporates disulfide bonds, since this form is sensitive to reducing agent. The C48A and C139A mutants of hPNMT, which are incapable of forming the disulfide bond observed in the crystal structure, have a decreased propensity to form dimer in solution. Those dimers that do form are also sensitive to reducing agent. Further, the C48A/C139A double mutant shows only monomeric behaviour. Both dimeric and monomeric hPNMT, as well as mutants have wildtype enzyme activity. These results show that a variety of disulfides, including those observed in the crystal structure, can form in solution. In addition, disulfide-linked dimers are as active as the monomeric enzyme indicating that the crystal structure of the protein is a valid target for inhibitor design. PMID- 15893507 TI - Conserved protein TTHA1554 from Thermus thermophilus HB8 binds to glutamine synthetase and cystathionine beta-lyase. AB - TTHA1554 was found as a hypothetical protein composed of 95 amino acids in the genome of the extremely thermophilic bacterium, Thermus thermophilus HB8. Proteins homologous to TTHA1554 are conserved in several bacteria and archaea, although their functions are unknown. To investigate the function of TTHA1554, we identified interacting proteins by using a pull-down assay and mass spectrometry. TTHA1329, which is glutamine synthetase, and TTHA1620, a putative aminotransferase, were identified as TTHA1554 binding proteins. The interactions with TTHA1329 and TTHA1620 were validated using in vitro pull-down assays and surface plasmon resonance biosensor assays with recombinant proteins. Since sequence homology analyses suggested that TTHA1620 was a pyridoxal 5'-phosphate dependent enzyme, such as an aminotransferase, a cystathionine beta-lyase or a cystalysin, putative substrates were investigated. When cystathionine, cystine and S-methylcysteine were used as substrates, pyruvate was produced by TTHA1620. The data revealed that TTHA1620 has cystathionine beta-lyase enzymatic activity. When TTHA1554 was added to the reaction mixtures, the glutamine synthetase and cystathionine beta-lyase enzymatic activities both increased by approximately two fold. These results indicated that TTHA1554 is a novel protein (we named it GCBP: glutamine synthetase and cystathionine beta-lyase binding protein) that binds to glutamine synthetase and cystathionine beta-lyase. PMID- 15893508 TI - Practice of adult liver transplantation in Italy. Recommendations of the Italian Association for the Study of the Liver (A.I.S.F.). AB - Liver transplantation is an efficient procedure as performed in Italy, yet major differences are present in terms of practice. In an effort to facilitate an homogeneous practice of liver transplantation in Italy, the Italian Association for the Study of Liver Disease has instituted a Commission aimed at providing recommendations on non-urgent liver transplantation in adults, based on current evidence. This nation-wide commission which included experienced hepatologists, surgeons and pathologists with major interest in liver transplantation has drafted a final document in October 2004, approved by the Italian Association for the Study of Liver Governing Board, whose key arguments and main conclusions are summarised in the present paper. The Commission has made specific recommendations on the following topics: the current needs of liver transplantation in Italy; the indications to liver transplantation and re-liver transplantation, with special reference to controversial issues and the minimal listing criteria; the use of marginal donors and the need to optimise donor/recipient matching; the use of living donor liver transplantation; the management of the waiting list and the introduction of Model for End-Stage Liver Disease to define priorities; the clinical management of liver transplantation recipients and disease recurrence; the implementation of audits and outcome monitoring; the training of transplant surgeons and hepatologists and the requirements for Centre accreditation; the pathology of liver transplantation. PMID- 15893509 TI - Environmental factors that enhance the action of the cell penetrating peptide pep 1 A spectroscopic study using lipidic vesicles. AB - Pep-1 is a cell penetrating peptide (CPP) derived from the nuclear localization sequence of Simian Virus 40 large antigen T and from reverse transcriptase of Human Immunodeficiency Virus. Although it has been successfully used to transport proteins into cells, its action at the molecular level is not yet clear, mainly the local environmental factors that condition partition and translocation. Characterization in aqueous medium and quantification of partition into bilayers were carried out. Dynamic light scattering studies show that pep-1 self associates in aqueous medium. The role of the bilayer phase, anionic lipids, ionic strength of the medium, reducing agents and pep-1 concentration on the extent and kinetics of partition were studied. Unlike others cationic CPP (e.g. penetratin) pep-1 has a high affinity to neutral vesicles (Kp = 2.8 x 10(3)), which is enhanced by anionic lipids. In a reduction environment partition is strongly inhibited (Kp = 2.2 x 10(2)), which might be a key-feature in the biological action of pep-1. Peptide incorporation takes place in the millisecond time-range to the lipidic interfaces. These environmental factors are systematized to enlighten how they help cellular uptake. PMID- 15893510 TI - Protein mediated glycolipid transfer is inhibited FROM sphingomyelin membranes but enhanced TO sphingomyelin containing raft like membranes. AB - The mammalian glycolipid transfer protein, GLTP, catalyzes the transfer in vitro of glycolipids between membranes. In this study we have examined on one hand the effect of the variations in the donor vesicle composition and on the other hand the effects of variations in the acceptor vesicle composition on the GLTP catalyzed transfer kinetics of galactosylceramide between bilayer vesicles. For this purpose a resonance energy transfer assay was used, the energy donor being anthrylvinyl-galactosylceramide and the energy acceptor DiO-C16. First, we show that the transfer of anthrylvinyl-galactosylceramide from palmitoyl-oleoyl phosphatidylcholine donor vesicles was faster than from dipalmitoyl phosphatidylcholine vesicles, and that there is no transfer from palmitoyl sphingomyelin vesicles regardless of the cholesterol amount. In this setup the acceptor vesicles were always 100% palmitoyl-oleoyl-phosphatidylcholine. We also showed that the transfer in general is faster from small highly curved vesicles compared to that from larger vesicles. Secondly, by varying the acceptor vesicle composition we showed that the transfer is faster to mixtures of sphingomyelin and cholesterol compared to mixtures of phosphatidylcholines and cholesterol. Based on these experiments we conclude that the GLTP mediated transfer of anthrylvinyl-galactosylceramide is sensitive to the matrix lipid composition and membrane bending. We postulate that a tightly packed membrane environment is most effective in preventing GLTP from accessing its substrates, and cholesterol is not required to protect the glycosphingolipid in the membrane from being transferred by GLTP. On the other hand GLTP can more easily transfer glycolipids to 'lipid raft' like membranes, suggesting that the protein could be involved in raft assembly. PMID- 15893511 TI - The role of cysteine residues in the sulphate transporter, SHST1: construction of a functional cysteine-less transporter. AB - We investigated the role of cysteine residues in the sulphate transporter, SHST1, with the aim of generating a functional cysteine-less variant. SHST1 contains five cysteine residues and none was essential for function. However, replacement of C421 resulted in a reduction in transport activity. Sulphate transport by C205 mutants was dependent on the size of the residue at this position. Alanine at position 205 resulted in a complete loss of function whereas leucine resulted in a 3-fold increase in sulphate transport relative to wild type SHST1. C205 is located in a putative intracellular loop and our results suggest that this loop may be important for sulphate transport. By replacing C205 with leucine and the other four cysteine residues with alanine, we constructed a cysteine-less variant of SHST1 that has transport characteristics indistinguishable from wild type. This construct will be useful for further structure and function studies of SHST1. PMID- 15893512 TI - Internalisation of cell-penetrating peptides into tobacco protoplasts. AB - Cells are protected from the surrounding environment by plasma membrane which is impenetrable for most hydrophilic molecules. In the last 10 years cell penetrating peptides (CPPs) have been discovered and developed. CPPs enter mammalian cells and carry cargo molecules over the plasma membrane with a molecular weight several times their own. Known transformation methods for plant cells have relatively low efficiency and require improvement. The possibility to use CPPs as potential delivery vectors for internalisation in plant cells has been studied in the present work. We analyse and compare the uptake of the fluorescein-labeled CPPs, transportan, TP10, penetratin and pVEC in Bowes human melanoma cells and Nicotiana tabacum cultivar (cv.) SR-1 protoplasts (plant cells without cell wall). We study the internalisation efficiency of CPPs with fluorescence microscopy, spectrofluorometry and fluorescence-activated cell sorter (FACS). All methods indicate, for the first time, that these CPPs can internalise into N. tabacum cv. SR-1 protoplasts. Transportan has the highest uptake efficacy among the studied peptides, both in mammalian cells and plant protoplast. The internalisation of CPPs by plant protoplasts may open up a new effective method for transfection in plants. PMID- 15893513 TI - Ras couples phosphoinositide 3-OH kinase to the epithelial Na+ channel. AB - Aldosterone induces the expression of the small G protein K-Ras. Both K-Ras and its 1st effector phosphoinositide 3-OH kinase (PI3-K) are necessary and sufficient for the activation of ENaC increasing channel open probability. The cell signaling mechanism by which K-Ras enhances ENaC activity, however, is uncertain. We demonstrate here that K-Ras significantly activates human ENaC reconstituted in Chinese hamster ovary cells approximately 3-fold. Activation in response to K-Ras was sensitive to the irreversible PI3-K inhibitor wortmannin but not the competitive LY294002 inhibitor of this phospholipid kinase. Similarly, a PI3-K 1st effector-specific Ras mutant (G12:C40) enhanced ENaC activity in a wortmannin but not LY294002 sensitive manner. Constitutively active PI3-K also enhanced ENaC activity but in a wortmannin and LY294002 sensitive manner with the effects of PI3-K and K-Ras not being additive. The activation of ENaC by PI3-K was also sensitive to intracellular GDPbetaS. Constitutively active PI3-K that is incapable of interacting with K-Ras (K227E p110alpha) acted as dominant negative with respect to the regulation of ENaC even in the presence of K-Ras. K-Ras is known to directly interact with PI3-K with aldosterone promoting this interaction. Here we demonstrate that K-Ras also interacts with ENaC through an, as yet, undetermined mechanism. We conclude that K-Ras enhances ENaC activity by localizing PI3-K near the channel and stimulating of PI3-K activity. PMID- 15893514 TI - Precision parameters from spin-probe studies of membranes using a partitioning technique. application to two model membrane vesicles. AB - A new version of the ESR spin probe partitioning method is developed and applied to the study of hydration properties of dimyristoyl-phosphatidylglycerol (DMPG) and dimyristoyl-phosphatidylcholine (DMPC) vesicles as functions of salt concentration and temperature above the lipid phase transition. The small spin probe di-tert-butyl nitroxide (DTBN) is used in order to achieve motionally narrowed Electron Spin Resonance (ESR) spectra which may be analyzed with high precision. The new method relies on the use of the second harmonic display of the ESR spectrum followed by spectral line fitting. Spectral fitting yields precise ESR parameters giving detailed information on the surroundings of the spin probe in both phospholipid and aqueous phases. The nitrogen hyperfine coupling constant of DTBN arising from those probes occupying the vesicles is used to study the hydration of the vesicle surface. The hydration properties of the negatively charged vesicle surface of DMPG vesicles are affected by the addition of salt at all temperatures. In contrast, the hydration of DMPC vesicles does not change with salt concentration at the low temperatures. However, at higher temperatures the hydration properties of DMPC vesicle are affected by salt which is interpreted to be due to the faster motion of the phospholipid molecules. The partitioning of the spin probe increases with salt concentration for both DMPG and DMPC vesicles, while water penetration decreases simultaneously. The spin probe in the phospholipid bilayer exhibits anisotropic motion and the extent of the anisotropy is increased at the higher salt concentrations. PMID- 15893515 TI - Structural rearrangement of model membranes by the peptide antibiotic NK-2. AB - We have developed a novel alpha-helical peptide antibiotic termed NK-2. It efficiently kills bacteria, but not human cells, by membrane destruction. This selectivity could be attributed to the different membrane lipid compositions of the target cells. To understand the mechanisms of selectivity and membrane destruction, we investigated the influence of NK-2 on the supramolecular aggregate structure, the phase transition behavior, the acyl chain fluidity, and the surface charges of phospholipids representative for the bacterial and the human cell cytoplasmic membranes. The cationic NK-2 binds to anionic phosphatidylglycerol liposomes, causing a thinning of the membrane and an increase in the phase transition temperature. However, this interaction is not solely of electrostatic but also of hydrophobic nature, indicated by an overcompensation of the Zeta potential. Whereas NK-2 has no effect on phosphatidylcholine liposomes, it enhances the fluidity of phosphatidylethanolamine acyl chains and lowers the phase transition enthalpy of the gel to liquid cristalline transition. The most dramatic effect, however, was observed for the lamellar/inverted hexagonal transition of phosphatidylethanolamine which was reduced by more than 10 degrees C. Thus, NK-2 promotes a negative membrane curvature which can lead to the collapse of the phosphatidylethanolamine-rich bacterial cytoplasmic membrane. PMID- 15893516 TI - High zinc sensitivity and pore formation in an invertebrate P2X receptor. AB - To investigate fast purinergic signaling in invertebrates, we examined the functional properties of a P2X receptor subunit cloned from the parasitic platyhelminth Schistosoma mansoni. This purinoceptor (SmP2X) displays unambiguous homology of primary sequence with vertebrate P2X subunits. SmP2X subunits assemble into homomeric ATP-gated channels that exhibit slow activation kinetics and are blocked by suramin and PPADS but not TNP-ATP. SmP2X mediates the uptake of the dye YO-PRO-1 through the formation of large pores and can be blocked by submicromolar concentrations of extracellular Zn2+ ions (IC50 = 0.4 microM). The unique receptor phenotype defined by SmP2X suggests that slow kinetics, modulation by zinc and the ability to form large pores are ancestral properties of P2X receptors. The high sensitivity of SmP2X to zinc further reveals a zinc regulation requirement for the parasite's physiology that could potentially be exploited for therapeutic purposes. PMID- 15893518 TI - Stabilized plasmid-lipid particles containing PEG-diacylglycerols exhibit extended circulation lifetimes and tumor selective gene expression. AB - Stabilized plasmid lipid particles (SPLP) consist of a single copy of DNA surrounded by a lipid bilayer. The particles are small ( approximately 100 nm), stable, monodisperse and have a low surface charge. A diffusible polyethylene glycol (PEG) coating attached to a lipid anchor is critical to the SPLP's functionality. The PEG-lipid exchanges out of the bilayer at a rate determined by the size of the lipid anchor. Here we show that SPLP can be prepared using a series of PEG-diacylglycerol lipids (PEG-S-DAGs). SPLP were prepared incorporating PEG-dimyristoylglycerol (C14), PEG-dipalmitoylglycerol (C16) or PEG distearoylglycerol (C18) and the rate of PEG-lipid diffusion from the bi-layer determined using a FRET assay. SPLP pharmacokinetics confirm a correlation between the stability of the PEG-lipid component and circulation lifetime. PEG-S DAGs with longer lipid anchors yield more stable SPLP particles with longer circulation half-lives yielding an increase in tumor delivery and gene expression. PEG-distearoylglycerol (C18) containing SPLP bypass so-called 'first pass' organs, including the lung, and elicit levels of gene expression in distal tumor tissue 100- to 1000-fold greater than that observed in any other tissue. The incorporation of PEG-S-DAG in SPLP confirms that small size, low surface charge and extended circulation lifetimes are prerequisite to the accumulation and tumor selective expression of plasmid DNA following systemic administration. PMID- 15893517 TI - Nectin-like molecule 1 is a protein 4.1N associated protein and recruits protein 4.1N from cytoplasm to the plasma membrane. AB - Nectins are immunoglobulin superfamily adhesion molecules that participate in the organization of epithelial and endothelial junctions. Sharing high homology with the poliovirus receptor (PVR/CD155), nectins were also named poliovirus receptor related proteins (PRRs). Four nectins and five nectin-like molecules have been identified. Here we describe the cloning and characterization of human and mouse nectin-like molecular 1 (NECL1). Human and mouse NECL1 share 87.3% identity at the amino acid level. NECL1 contains an ectodomain made of three immunoglobulin like domains, and a cytoplasmic region homologous to those of glycophorin C and contactin-associated protein. RNA blot and in situ hybridization analysis showed that NECL1 predominantly expressed in the central nervous system, mainly in neuronal cell bodies in a variety of brain regions including the cerebellum, cerebral cortex and hippocampus. In vitro binding assay proved the association of NECL1 with protein 4.1N. NECL1 localizes to the cell-cell junctions and recruits protein 4.1N to the plasma membranes through its C-terminus, thus may regulate the function of the cell-cell junction. We propose that the NECL1 and protein 4.1N complex is involved in the morphological development, stability, and dynamic plasticity of the nervous system. PMID- 15893519 TI - Changes in the morphology of cell-size liposomes in the presence of cholesterol: formation of neuron-like tubes and liposome networks. AB - Spontaneous changes in the morphology of cell-size liposomes (dioleoylphosphatidylcholine, DOPC and egg PC) as model cells were investigated in the presence of cholesterol. Tube structures and liposome networks connected by the tubes were observed in the presence of 5-30% cholesterol by dark-field and laser-scanning microscopy. Furthermore, in the presence of more than 40 mol% of cholesterol, the tubes disappeared and changed to small liposomes. Thus, cholesterol induced a morphological change in giant liposomes from tubes to small liposomes. These phenomena may be related to the role of cholesterol in the morphological changes in living cells such as neurons. PMID- 15893520 TI - Phospholipid diversity: correlation with membrane-membrane fusion events. AB - The transport of various metabolically important substances along the endocytic and secretory pathways involves budding as well as fusion of vesicles with various intracellular compartments and plasma membrane. The membrane-membrane fusion events between various sub-compartments of the cell are believed to be mainly mediated by so-called "fusion proteins". This study shows that beside the proteins, lipid components of membrane may play an equally important role in fusion and budding processes. Inside out (ISO) as well as right side out (RSO) erythrocyte vesicles were evaluated for their fusogenic potential using conventional membrane fusion assay methods. Both fluorescence dequenching as well as content mixing assays revealed fusogenic potential of the erythrocyte vesicles. Among two types of vesicles, ISO were found to be more fusogenic as compared to the RSO vesicles. Interestingly, ISO retained nearly half of their fusogenic properties after removal of the proteins, suggesting the remarkable role of lipids in the fusion process. In another set of experiments, fusogenic properties of the liposomes (subtilosome), prepared from phospholipids isolated from Bacillus subtilis (a lower microbe) were compared with those of erythrocyte vesicles. We have also demonstrated that various types of vesicles upon interaction with macrophages deliver encapsulated materials to the cytosol of the cells. Membrane-membrane fusion was also followed by the study, in which a protein synthesis inhibitor ricin A (that does not cross plasma membrane), when encapsulated in the erythrocyte vesicles or subtilosomes was demonstrated to gain access to the cytosol. PMID- 15893521 TI - Agonist-induced up-regulation of human somatostatin receptor type 1 is regulated by beta-arrestin-1 and requires an essential serine residue in the receptor C tail. AB - We have previously shown that the human somatostatin receptor type 1 (hSSTR1) does not undergo agonist-induced internalization, but is instead up-regulated at the membrane upon prolonged somatostatin (SST) exposure. The deletion of the carboxyterminal C-tail of the receptor completely abolishes up-regulation. To identify molecular signals that mediate hSSTR1 up-regulation, we created mutant receptors with progressive C-tail deletions. Up-regulation was found to be absent in mutants lacking residues Lys359-Ser360-Arg361. Moreover, point mutation of Ser360 to Ala completely abolished up-regulation. The coexpression of wild type hSSTR1 with V53D, a dominant negative mutant of beta-arrestin-1, completely blocked hSSTR1 up-regulation. Further analysis demonstrated that calcium calmodulin (CaM) dependent kinases were essential for the SST-induced up regulation response. Like wild type receptors, all mutants failed to internalize after agonist exposure and were able to inhibit forskolin-stimulated cAMP accumulation. Taking these data together, we suggest that SST-induced hSSTR1 up regulation is critically dependent upon a specific Lys-Ser-Arg sequence in the C tail of the receptor, with Ser360 being essential. Up-regulation also requires the participation of CaM protein kinases and interactions with beta-arrestins. In contrast, coupling to adenyl cyclase (AC) and internalization occur independently of molecular signals in the receptor's C-tail. PMID- 15893522 TI - New effects in polynucleotide release from cationic lipid carriers revealed by confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking. AB - We report on new insights into the mechanisms of short single and double stranded oligonucleotide release from cationic lipid complexes (lipoplexes), used in gene therapy. Specifically, we modeled endosomal membranes using giant unilamellar vesicles and investigated the roles of various individual cellular phospholipids in interaction with lipoplexes. Our approach uses a combination of confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking, revealing several new aspects of the release: (a) phosphatidylserine and phosphatidylethanolamine are equally active in disassembling lipoplexes, while phosphatidylcholine and sphingomyelin are inert; (b) in contrast to earlier findings, phosphatidylethanolamine alone, in the absence of anionic phosphatidylserine triggers extensive release; (c) a double-stranded DNA structure remains well preserved after release; (d) lipoplexes exhibited preferential binding to transient lipid domains, which appear at the onset of lipoplex attachment to originally uniform membranes and vanish after initiation of polynucleotide release. The latter effect is likely related to phosphatidyleserine redistribution in membranes due to lipoplex binding. Real time tracking of single DOTAP/DOPE and DOTAP/DOPC lipoplexes showed that both particles remained compact and associated with membranes up to 1-2 min before fusion, indicating that a more complex mechanism, different from suggested earlier rapid fusion, promotes more efficient transfection by DOTAP/DOPE complexes. PMID- 15893523 TI - Relational language and the development of relational mapping. AB - We test the claim that learning and using language for spatial relations can influence spatial representation and reasoning. Preschool children were given a mapping task in which they were asked to find a "winner" placed in a three-tiered box after seeing one placed in a virtually identical box. The correct choice was determined by finding the corresponding relative location in the test box, making it a difficult task for preschool children. We found that hearing language for spatial relations facilitated children's mapping performance. We found effects at younger ages on easier tasks (Experiments 1 and 2) and at older ages on harder tasks (Experiment 3). The effects of spatial relational language differed predictably according to the semantics of the terms children heard (Experiment 4). Finally, the effects of spatial language were maintained over time (Experiment 5): children given one initial exposure to the spatial terms maintained their advantage over baseline children when they again carried out the mapping task 2 days later, with no further exposure to the spatial terms. The evidence is consistent with the explanation that language bolsters children's spatial encodings, which in turn supports their mapping performance. PMID- 15893524 TI - Infants use meter to categorize rhythms and melodies: implications for musical structure learning. AB - Little is known about whether infants perceive meter, the underlying temporal structure of music. We employed a habituation paradigm to examine whether 7-month old infants could categorize rhythmic and melodic patterns on the basis of the underlying meter, which was implied from event and accent frequency of occurrence. In Experiment 1, infants discriminated duple and triple classes of rhythm on the basis of implied meter. Experiment 2 replicated this result while controlling for rhythmic grouping structure, confirming that infants perceived metrical structure despite occasional ambiguities and conflicting group structure. In Experiment 3, infants categorized melodies on the basis of contingencies between metrical position and pitch. Infants presented with metrical melodies detected reversals of pitch/meter contingencies, while infants presented with non-metrical melodies showed no preference. Results indicate that infants can infer meter from rhythmic patterns, and that they may use this metrical structure to bootstrap their knowledge acquisition in music learning. PMID- 15893525 TI - Color categories: evidence for the cultural relativity hypothesis. AB - The question of whether language affects our categorization of perceptual continua is of particular interest for the domain of color where constraints on categorization have been proposed both within the visual system and in the visual environment. Recent research (Roberson, Davies, & Davidoff, 2000; Roberson et al., in press) found substantial evidence of cognitive color differences between different language communities, but concerns remained as to how representative might be a tiny, extremely remote community. The present study replicates and extends previous findings using additional paradigms among a larger community in a different visual environment. Adult semi-nomadic tribesmen in Southern Africa carried out similarity judgments, short-term memory and long-term learning tasks. They showed different cognitive organization of color to both English and another language with the five color terms. Moreover, Categorical Perception effects were found to differ even between languages with broadly similar color categories. The results provide further evidence of the tight relationship between language and cognition. PMID- 15893527 TI - MR imaging of articular cartilage: current state and recent developments. AB - Osteoarthritis is the most common type of arthritis and a frequent cause of pain and disability. A number of exciting surgical treatment modalities have been introduced recently, including autologous chondrocyte transplantation and osteochondral allografting or autografting. MR imaging offers the distinct advantage of visualizing the articular cartilage directly. MR imaging can detect signal and morphologic changes in the cartilage and has been used to detect cartilage surface fraying, fissuring, and varying degrees of cartilage thinning. PMID- 15893528 TI - New techniques for cartilage imaging: T2 relaxation time and diffusion-weighted MR imaging. AB - In view of recent therapeutic approaches to cartilage damage in osteoarthritis, it is necessary to develop and further refine noninvasive quantitative tools for specific diagnosis and follow-up studies. There is considerable experimental and some clinical experience with T2 relaxation time measurements. Motivation for diffusion-weighted imaging and diffusion-tensor imaging as comparably new techniques for cartilage imaging is to obtain directly additional three dimensional architectural and directional information about the cartilage matrix. PMID- 15893529 TI - MR imaging of epiphyseal lesions of the knee: current concepts, challenges, and controversies. AB - This article focuses on spontaneous painful conditions involving the subchondral bone and marrow of mature knee epiphyses. MR imaging is the technique of choice for the work-up of these lesions and enables distinction of two main categories of lesions on the basis of T1-weighted images: avascular necrosis, and lesions presenting the bone marrow edema pattern. This latter category encompasses spontaneous osteonecrosis of the knee, and a variety of self-resolving conditions that may be differentiated by the study of the subchondral bone marrow area on T2 weighted images. Behind definite appellation of lesions, the challenge for the radiologist is to provide a prognosis: the distinction between self-resolving lesions from those that may evolve to epiphyseal collapse and joint impairment should be possible in most cases. PMID- 15893530 TI - High- versus low-field MR imaging. AB - The role of MR imaging as a noninvasive technique in the detection and evaluation of musculoskeletal diseases is unquestionable. Most of the studies reported in the literature are based on high-field MR imaging. Initial studies performed with low-field-strength have reported unsatisfactory results in the assessment of the musculoskeletal system. Recent improvements, however, have generated a renewed interest in low-field-strength MR imaging. This article presents the principal applications and results published in the literature. PMID- 15893531 TI - Shoulder MR arthrography: how, why, when. AB - This article reviews current MR techniques for shoulder imaging, discusses advantages and disadvantages of each, and reviews the literature regarding sensitivity, specificity, and accuracy of MR arthrography versus nonenhanced MR in the evaluation of shoulder pathology, specifically, glenoid labral tears and rotator cuff tears. PMID- 15893532 TI - Ankle MR arthrography: how, why, when. AB - MR arthrography has become an important tool for the assessment of a variety of ankle disorders. MR arthrography may facilitate the evaluation of patients with suspected intra-articular pathology in whom conventional MR imaging is not sufficient for an adequate diagnosis and be useful for therapy planning. MR arthrography is valuable in the evaluation of ligamentous injuries, impingement syndromes, cartilage lesions, osteochondral lesions of the talus, loose bodies, and several synovial joint disorders. Indirect MR arthrography is a useful adjunct to conventional MR imaging and may be preferable to direct MR arthrography in cases in which an invasive procedure is contraindicated or when fluoroscopy is not available. PMID- 15893533 TI - Wrist MR arthrography: how, why, when. AB - MR imaging of the wrist frequently represents a diagnostic challenge for radiologists because of the complex anatomy of this joint, small size of its components, and little known pathologic conditions. MR arthrography combines the advantages of conventional MR imaging and arthrography by improving the visualization of small intra-articular abnormalities. This article reviews the current role of MR arthrography in the evaluation of wrist joint disorders considering the relevant aspects of anatomy, techniques, and applications. PMID- 15893534 TI - MR arthrography of the knee: how, why, when. AB - MR arthrography combines the techniques of arthrography with MR imaging to benefit from the added imaging information afforded by intra-articular distention. This article reviews technical considerations for MR arthrography, potential complications, indications, pitfalls in imaging diagnosis, and commonly encountered pathology. It is an elegant study that can offer precise diagnostic information in the appropriate clinical setting. PMID- 15893535 TI - MR imaging of the diabetic foot: diagnostic challenges. AB - Pedal complications of diabetes have long presented a challenge for the clinician and radiologist predominately related to the difficulty in distinguishing infection from neuroarthropathy. The spectrum of diabetic foot infections is broad, ranging from callous and ulcer formation, to septic arthritis, abscess formation, and osteomyelitis. This article summarizes the MR imaging findings in the diabetic foot and the optimal pulse sequences. Focus is placed on aids in differentiating diabetic infection from other entities and increasing the specificity of diagnosing diabetic foot complications. PMID- 15893536 TI - Normal variants and frequent marrow alterations that simulate bone marrow lesions at MR imaging. AB - MR imaging of the spine is routinely performed for the assessment of patients with spine-related symptoms and of patients with cancer. This article addresses normal variants and frequent alterations of the vertebral bone marrow that are encountered on MR imaging studies and can simulate lesions. PMID- 15893537 TI - The complementary roles of MR imaging and ultrasound of tendons. AB - The choice to use MR imaging or ultrasound to depict tendon pathology has traditionally depended on the imager's level of experience and comfort with the modality, and less so the individual strengths of either modality. Although this may be an acceptable rationale, it does not fully take advantage of the strength of either modality or the potential benefits of combining both modalities. This article demonstrates the complementary roles of these two modalities through a variety of clinical examples, based on experience working in a subspecialty hospital dedicated to orthopedic and rheumatologic diseases. PMID- 15893538 TI - Inhibition of leukocyte function and interleukin-2 gene expression by 2 methylarachidonyl-(2'-fluoroethyl)amide, a stable congener of the endogenous cannabinoid receptor ligand anandamide. AB - Arachidonylethanolamide (anandamide, AEA) has been identified as an endogenous ligand for cannabinoid receptors CB1 and CB2. Characterization of the direct cannabimimetic actions of anandamide has been hampered by its short duration of action and rapid degradation in in vivo and in vitro systems to arachidonic acid, a precursor in the biosynthesis of a broad range of biologically active molecules. In the present studies, we utilized 2-methylarachidonyl-(2' fluoroethyl)amide (F-Me-AEA), an analog of anandamide resistant to enzymatic degradation, to determine whether F-Me-AEA modulated T cell function similar to that of plant-derived cannabinoids. Indeed, F-Me-AEA at low micromolar concentrations exhibited a marked inhibition of phorbol ester plus calcium ionophore (PMA/Io)-induced IL-2 protein secretion and steady state mRNA expression. Likewise, a modest suppression of the mixed lymphocyte response was observed in the presence of F-Me-AEA indicating an alteration in T cell responsiveness to allogeneic MHC class II antigens. F-Me-AEA was also found to modestly inhibit forskolin-stimulated adenylate cyclase activity in thymocytes and splenocytes, a hallmark of cannabinoid receptor agonists. Further characterization of the influence of F-Me-AEA on the cAMP signaling cascade revealed an inhibition of CREB-1/ATF-1 phosphorylation and subsequently, an inhibition of CRE DNA binding activity. Characterization of nuclear binding proteins further revealed that NF-AT and, to a lesser extent, NF-kappaB DNA binding activities were also suppressed. These studies demonstrate that F-Me-AEA modulates T cell function in a similar manner to plant-derived and endogenous cannabinoids and therefore can be utilized as an amidase- and hydrolysis resistant endogenous cannabinoid. PMID- 15893539 TI - Comparative metabolism of methacrylonitrile and acrylonitrile to cyanide using cytochrome P4502E1 and microsomal epoxide hydrolase-null mice. AB - Methacrylonitrile (MAN) and acrylonitrile (AN) are metabolized via glutathione (GSH) conjugation or epoxide formation. We have recently shown that CYP2E1 is essential for AN epoxidation and subsequent cyanide liberation. Current studies were designed to compare the enzymatic basis of MAN vs. AN metabolism to cyanide using wild-type (WT), CYP2E1-, and mEH-null mice. Mice received a single gavage dose of 0.047, 0.095, 0.19, or 0.38 mmol/kg of MAN or AN, and blood cyanide was measured at 1 or 3 h later. Blood cyanide levels in WT mice treated with AN or MAN were dose and time dependent. At equimolar doses, significantly higher levels of cyanide were detected in the blood of MAN- vs. AN-treated mice. Further, while significant reduction in blood cyanide levels occurred in MAN-treated CYP2E1-null vs. WT mice, AN metabolism to cyanide was largely abolished in CYP2E1-null mice. Pretreatment of mice with 1-aminobenzotriazole (ABT, CYP inhibitor) demonstrated that CYPs other than CYP2E1 also contribute to MAN metabolism to cyanide. Blood cyanide levels in mEH-null mice treated with aliphatic nitriles are generally lower than levels in similarly treated WT mice. Western blot analysis showed that expression of sEH was greater in male vs. female mice. The role of various epoxide hydrolases (EHs) in the production of cyanide from aliphatic nitriles is apparently structure and dose dependent. Regardless of genotype, significantly higher levels of cyanide were measured in the blood of male vs. female mice treated with MAN or AN. In conclusion, these data showed that (1) at equimolar doses, higher blood cyanide levels were detected in mice treated with MAN vs. AN; (2) while CYP2E1 is the only enzyme responsible for AN metabolism to cyanide, other CYPs also contribute to MAN metabolism; and (3) significantly higher levels of cyanide were measured in the blood of male vs. female treated with either nitrile. Higher blood cyanide levels in male vs. female mice and in MAN- vs. AN treated mice may explain the gender-related differences in the toxicity of these chemicals and the greater potency of MAN vs. AN. PMID- 15893540 TI - The effect of marimastat, a metalloprotease inhibitor, on allergen-induced asthmatic hyper-reactivity. AB - This pilot study was designed to assess whether a synthetic matrix metalloproteinase (MMP) inhibitor has anti-inflammatory properties in mild asthma. Tumor necrosis factor alpha (TNFalpha) has been shown to be an important cytokine in the pathogenesis of allergic airway inflammatory responses, and its release can be inhibited by MMP inhibitors. Twelve atopic asthmatic subjects received the MMP inhibitor marimastat (5 mg) or placebo, twice daily for 3 weeks, separated by a 6-week washout period in a randomized, double-blind, cross-over manner. All subjects underwent an allergen inhalation provocation test to Dermatophagoides pteronyssinus before and after each study phase. Spirometry, exhaled NO (eNO) levels, differential sputum cell counts, an asthma symptom questionnaire, peak flow, and beta(2)-agonist usage were measured. Nine subjects completed the study, and, when compared with placebo, marimastat reduced bronchial hyper-responsiveness to inhaled allergen in these subjects from an allergen PC(20) of 22.2 AU/ml (95%CI 11.7-32.6) to 17.0 AU/ml (95%CI 7.6-26.4, P = 0.02). The marimastat phase showed a nonsignificant fall in sputum inflammatory cells. Marimastat did not modify eNO, FEV(1), asthma symptoms, or albuterol usage. In conclusion, airway responsiveness to allergen may be modified by a MMP inhibitor, perhaps via TNFalpha playing a role in airway inflammation and remodeling. PMID- 15893541 TI - Tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) induces cell proliferation in normal human bronchial epithelial cells through NFkappaB activation and cyclin D1 up-regulation. AB - Cigarette smoke contains several carcinogens known to initiate and promote tumorigenesis as well as metastasis. Nicotine is one of the major components of the cigarette smoke and the 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a tobacco-specific carcinogen. Here, we demonstrated that NNK stimulated cell proliferation in normal human bronchial epithelial cells (NHBE) and small airway epithelial cells (SAEC). Cells exposed to NNK resulted in an increase in the level of cyclin D1 protein (as early as 3-6 h). Increased phosphorylation of the Rb Ser(795) was detected at 6-15 h after NNK treatment and thereby promoted cells entering into the S phase (at 15-21 h). The increased cyclin D1 protein level was induced through activation of the transcription factor, nuclear factor kB (NFkappaB), in the NHBE cells. Treatment of the NHBE cells with PD98059, an ERK1/2 (extracellular signal-regulated protein kinase)-specific inhibitor, specifically suppressed the NNK-induced IkappaBalpha phosphorylation at position 32 of the serine residue, suggesting that the ERK1/2 kinase was involved in the IkappaBalpha phosphorylation induced by NFkappaB activation. To determine whether the NNK-induced NFkappaB activation and cyclin D1 induction were also observed in vivo, A/J mice were treated with NNK (9.1 mg) for 20 weeks and the results showed a significant induction of cyclin D1 and NFkappaB translocation determined by immunoblotting analyses. We further demonstrated that the nicotine acetylcholine receptor (nAchR), which contains the alpha3-subunit, was the major target mediating NNK-induced cyclin D1 expression in the NHBE cells. In summary, our findings demonstrate for the first time that NNK could stimulate normal human bronchial cell proliferation through activation of the NFkappaB, which in turn up regulated the cyclin D1 expression. PMID- 15893542 TI - Platelet-activating factor acetylhydrolase: selective inhibition by potent n alkyl methylphosphonofluoridates. AB - Platelet-activating factor (PAF) is a potent endogenous phospholipid modulator of diverse biological activities, including inflammation and shock. PAF levels are primarily regulated by PAF acetylhydrolases (PAF-AHs). These enzymes are candidate secondary targets of organophosphorus (OP) pesticides and related toxicants. Previously known OP inhibitors of other serine hydrolases were tested with PAF-AH from mouse brain and testes of established functional importance compared with the structurally different human plasma enzyme. Several key OP pesticides and their oxon metabolites were very poor inhibitors of mouse brain and human plasma PAF-AH in vitro but moderately active for mouse brain and blood PAF-AH in vivo (e.g., tribufos defoliant and profenofos insecticide, presumably following oxidative bioactivation). OP compounds were then designed for maximum in vitro potency and selectivity for mouse brain PAF-AH vs. acetylcholinesterase (AChE). Lead compounds were found in a series of benzodioxaphosphorin 2-oxides. Ultrahigh potency and selectivity were achieved with n-alkyl methylphosphonofluoridates (long-chain sarin analogs): mouse brain and testes IC50 < or = 5 nM for C(8)-C(18) analogs and 0.1-0.6 nM for C(13) and C(14) compounds; human plasma IC50 < or = 2 nM for C(13)-C(18) analogs. AChE inhibitory potency decreased as chain length increased with maximum brain PAF-AH/AChE selectivity (>3000-fold) for C(13)-C(18) compounds. The toxicity of i.p. administered PAF (LD50 ca. 0.5 mg/kg) was increased less than 2-fold by pretreatment with tribufos or the C(13)n-alkyl methylphosphonofluoridate. These studies with a mouse model indicate that PAF-AH is not a major secondary target of OP pesticide poisoning. The optimized PAF-AH inhibitors may facilitate investigations on other aspects of PAF metabolism and action. PMID- 15893543 TI - Formation of PAH-DNA adducts after in vivo and vitro exposure of rats and lung cells to different commercial carbon blacks. AB - OBJECTIVE: The current study was designed to test the possible release and bioavailability of polycyclic aromatic hydrocarbons (PAHs) from a set of commercial carbon blacks (CBs) as well as the ability of these PAHs to form bulky DNA adducts. METHODS: In four commercial CBs (Printex 90, Sterling V, N330, Lampblack 101), leaching of PAH was examined through (1) release of parent PAHs in saline with or without surfactant, and (2) PAH adducts in lung epithelial cells (A549) or in rat lungs after exposure to two CBs (Printex 90, Sterling V) for 13 weeks (50 mg/m(3)). In vitro experiments were done with original and extracted particles, as well as organic extracts of CB in DMSO. As positive controls, B[a]P (0.03 microM) and a mixture of 16 PAHs (0.1 microM) were used. RESULTS: No leaching of PAHs was measured in saline or surfactant-containing saline. In vitro incubations with CB particles (30-300 microg/cm(2)) revealed no adduct spots except for Sterling V. However, the spot was not concentration dependent and remains unidentified. Lung DNA from rats after inhalation of Printex 90 or Sterling V showed no spots related to PAH-DNA adduct formation compared to sham-exposed rats. CONCLUSION: The results suggest that PAHs are very tightly bound to these CBs. Only using organic extracts or particles of low surface Sterling V, with high PAH content, PAHs may become available to form PAH DNA adducts. However, the in vitro conditions showing this effect will not be encountered in vivo and renders this mechanism in particle-induced lung cancer at in vivo exposures highly unlikely. PMID- 15893544 TI - Increase in a distinct pulmonary macrophage subset possessing an antigen presenting cell phenotype and in vitro APC activity following silica exposure. AB - Silica inhalation results in chronic lung inflammation and fibrosis. While the role of the alveolar macrophage (AM) is considered key to the effects of silica on lung pathology, the etiology is not completely understood. Evidence suggests an increase in antigen presenting cell (APC) activity as a contributing factor to this process, as well as potential roles for both AM and interstitial macrophages (IM) in silicosis. In order to study the effects of crystalline silica on the APC activity of pulmonary macrophages, mice were exposed intranasally and changes in pulmonary macrophage populations were assessed using flow cytometry. Following intranasal instillation of silica, a significant increase in the APC activity of AM was observed, as well as a significant increase in a subset of IM expressing classic APC markers (MHC class II, CD11c). In addition, an in vitro system using bone marrow-derived macrophages (BMDM) was generated to assess the effects of silica on the APC activity of macrophages in vitro. Data using BMDM in the in vitro APC assay demonstrated a significant increase in APC activity following silica exposure, but not following exposure to saline or a control particle (TiO(2)). Using a combination of in vivo and in vitro experiments, the current study describes a significant increase in an interstitial macrophage subset with an APC phenotype, as well as an increase in the APC activity of both AM and BMDM, as a direct result of exposure to crystalline silica. These studies suggest a specific mechanism, macrophage subset activation, by which crystalline silica exposure results in chronic pulmonary inflammation and, eventually, fibrosis. PMID- 15893545 TI - Accumulation of methylmercury or polychlorinated biphenyls in in vitro models of rat neuronal tissue. AB - In vivo exposure levels for neurotoxicants are often reported in parts per million (ppm) concentration in tissue, whereas exposure levels in experiments utilizing in vitro models are most commonly reported in micromolar (muM) concentration in the exposure solution. The present experiments sought to determine whether or not in vitro solution concentration was an appropriate dose metric for comparison to in vivo tissue levels for lipophilic compounds. To do so, the accumulation of the polychlorinated biphenyl (PCB) mixture Aroclor 1254 (A1254) or methylmercury (MeHg) was examined in three commonly utilized in vitro neuronal tissue models: nerve growth factor differentiated pheochromocytoma (PC12) cells, primary cultures of rat neocortical cells, and adult rat hippocampal slices. Tissues were exposed to A1254 (0.65 ppm) or to MeHg (0.0033 0.33 ppm) in serum-free media for 1 or 24 h. Total PCB or mercury accumulation was measured by dual column gas chromatography with electron capture detection or by cold vapor atomic absorption, respectively. PC12 cells accumulated 66.7 and 103.8 ppm PCBs after 1 and 24 h exposure to A1254. Neocortical neurons also accumulated significant concentrations of PCBs, but less so than PC12 cells. After 1 h exposure to 0.65 ppm A1254, slices contained 3.46 and 0.81 ppm PCBs when exposed in a static and perfused system, respectively. After 1 h exposure to 0.0033, 0.033, and 0.33 ppm MeHg, PC12 cells contained 0.3, 2.2, and 17.7 ppm mercury, respectively; after 24 h, PC12 cells contained 0.4, 2.8, and 21.9 ppm. Hippocampal slices accumulated 1.7 and 4.8 ppm mercury after 1 and 3 h exposure to 0.33 ppm MeHg. For comparison, mercury accumulation in rat fetal and pup brain tissue after maternal exposure [0, 0.1, 1.0, or 2.0 mg/kg/day MeHg from gestational day (GD) 6-15] ranged from 0.05 to 7.89 ppm in 0.1 mg/kg dose animals on postnatal day 10 and 2.0 mg/kg dose animals on GD16, respectively. These results demonstrate that accumulation of PCBs and MeHg in vitro is tissue-, time , and concentration-dependent and indicates that tissue levels rather than exposure concentrations are a more appropriate metric for comparison of in vitro to in vivo effects. PMID- 15893546 TI - Alteration at translational but not transcriptional level of transferrin receptor expression following manganese exposure at the blood-CSF barrier in vitro. AB - Manganese exposure alters iron homeostasis in blood and cerebrospinal fluid (CSF), possibly by acting on iron transport mechanisms localized at the blood brain barrier and/or blood-CSF barrier. This study was designed to test the hypothesis that manganese exposure may change the binding affinity of iron regulatory proteins (IRPs) to mRNAs encoding transferrin receptor (TfR), thereby influencing iron transport at the blood-CSF barrier. A primary culture of choroidal epithelial cells was adapted to grow on a permeable membrane sandwiched between two culture chambers to mimic blood-CSF barrier. Trace (59)Fe was used to determine the transepithelial transport of iron. Following manganese treatment (100 microM for 24 h), the initial flux rate constant (K(i)) of iron was increased by 34%, whereas the storage of iron in cells was reduced by 58%, as compared to controls. A gel shift assay demonstrated that manganese exposure increased the binding of IRP1 and IRP2 to the stem loop-containing mRNAs. Consequently, the cellular concentrations of TfR proteins were increased by 84% in comparison to controls. Assays utilizing RT-PCR, quantitative real-time reverse transcriptase-PCR, and nuclear run off techniques showed that manganese treatment did not affect the level of heterogeneous nuclear RNA (hnRNA) encoding TfR, nor did it affect the level of nascent TfR mRNA. However, manganese exposure resulted in a significantly increased level of TfR mRNA and reduced levels of ferritin mRNA. Taken together, these results suggest that manganese exposure increases iron transport at the blood-CSF barrier; the effect is likely due to manganese action on translational events relevant to the production of TfR, but not due to its action on transcriptional, gene expression of TfR. The disrupted protein-TfR mRNA interaction in the choroidal epithelial cells may explain the toxicity of manganese at the blood-CSF barrier. PMID- 15893547 TI - Coupled map lattice approximations for spatially explicit individual-based models of ecology. AB - Spatially explicit individual-based models are widely used in ecology but they are often difficult to treat analytically. Despite their intractability they often exhibit clear temporal and spatial patterning. We demonstrate how a spatially explicit individual-based model of scramble competition with local dispersal can be approximated by a stochastic coupled map lattice. The approximation disentangles the deterministic and stochastic element of local interaction and dispersal. We are thus able to understand the individual-based model through a simplified set of equations. In particular, we demonstrate that demographic noise leads to increased stability in the dynamics of locally dispersing single-species populations. The coupled map lattice approximation has general application to a range of spatially explicit individual-based models. It provides a new alternative to current approximation techniques, such as the method of moments and reaction-diffusion approximation, that captures both stochastic effects and large-scale patterning arising in individual-based models. PMID- 15893548 TI - A Michaelis-Menten-style model for the autocatalytic enzyme prostaglandin H synthase. AB - Prostaglandin H synthase (PGHS) is an autocatalytic enzyme which plays a key role in the arachidonic acid metabolic pathway. PGHS mediates the formation of prostaglandin H2, the precursor for a number of prostaglandins which are important in a wide variety of biological processes, including inflammation, blood clotting, renal function, and tumorigenesis. Here we present a Michaelis Menten-style model for PGHS. A stability analysis determines when the reaction becomes self-sustaining, and can help explain the regulation of PGHS activity in vivo. We also consider a quasi-steady-state approximation (QSSA) for the model, and present conditions under which the QSSA is expected to be a good approximation. Applying the QSSA for this model can be useful in computationally intensive modeling endeavors involving PGHS. PMID- 15893549 TI - Nonlinear waves in double-stranded DNA. AB - We propose a nonlinear model derived from first principles, to describe bubble dynamics of DNA. Our model equations include a term derived from the dissipative effect of intermolecular vibrational modes. Such modes are excited by the propagating bubble, and we term this 'curvature dissipation'. The equations that we derive allow for stable pinned localized kinks which form the bubble. We perform the stability analysis and specify the energy requirements for the motion of the localized solutions. Our findings are consistent with properties of DNA dynamics, and can be used in models for denaturation bubbles, RNA and DNA transcription, nucleotide excision repair and meiotic recombination. PMID- 15893550 TI - Checkpoint method for choice recovery in dynamic programming. AB - Many dynamic programming algorithms consist of a 'forward pass' computation to optimize a cost function, followed by a 'choice recovery' computation to construct a configuration that optimizes the cost function. 'Checkpointing' is a method to perform choice recovery using limited storage. During a forward pass, checkpoints in an optimal configuration are identified. Then recursion is used to fill in the portions of an optimal configuration between successive checkpoints. Choosing the number of checkpoints to use mediates a tradeoff between storage and speed. PMID- 15893551 TI - Sublinear growth of information in DNA sequences. AB - We introduce a novel method to analyse complete genomes and recognise some distinctive features by means of an adaptive compression algorithm, which is not DNA-oriented, based on the Lempel-Ziv scheme. We study the Information Content as a function of the number of symbols encoded by the algorithm and we analyse the dictionary created by the algorithm. Preliminary results are shown concerning regions showing a sublinear type of information growth, which is strictly connected to the presence of highly repetitive subregions that might be supposed to have a regulatory function within the genome. PMID- 15893552 TI - A decrease in drug resistance levels of the HIV epidemic can be bad news. AB - Transient decreases in the proportion of individuals newly infected with an HIV resistant virus (primary resistance) are documented for several cities of North America, including San Francisco. Using a staged SI deterministic model, we identified three potential causes consistent with the history of the epidemic: (1) increase in risky behaviour, (2) reduction in the proportion of HIV-acutely infected individuals undergoing treatment, and (3) replacement of mono- and dual drug therapies with triple-drug therapies. Although observed patterns resemble scenario 1 most closely, these explanations are not mutually exclusive and may have contributed synergistically to the decline. Under scenario 1 the counterintuitive situation arises where, although the proportion of primary resistance cases decreases transiently, the epidemic worsens because the actual numbers of infected individuals and of drug resistance carriers increases. Our results call for improved efforts to control the epidemic in developed nations, and highlight the usefulness of drug resistant strains as epidemiological markers. PMID- 15893553 TI - Drug resistance in an immunological model of HIV-1 infection with impulsive drug effects. AB - We consider an SIR-type model of immunological behaviour for HIV dynamics, including the effects of reverse transcriptase inhibitors and other drugs which prevent cellular infection. We use impulsive differential equations to model drug behaviour. We classify different regimes according to whether the drug efficacy is negligible, intermediate or high. We consider two strains of the virus: a wild type strain that can be controlled by both intermediate and high drug concentrations, and a mutant strain that can only be controlled by high drug concentrations. Drug regimes may take trajectories through one, two or all three regimes, depending on the dosage and the dosing schedule. We demonstrate that drug resistance arises at both intermediate and high drug levels. At low drug levels resistance does not emerge, but the total T cell count is proven to be significantly lower than in the disease-free state. At intermediate drug levels, drug resistance is guaranteed to emerge. At high drug levels, either the drug resistant strain will dominate or, in the absence of longer-lived reservoirs of infected cells, both viral sub-populations will be cleared. In the latter case the immune system is maintained by a population of T cells which have absorbed sufficient quantities of the drug to prevent infection by even the drug-resistant strain. We provide estimates of a range of dosages and dosing schedules which would, if physiologically tolerable, theoretically eliminate free virus in this system. Our results predict that to control viral load, decreasing the interval between doses is more effective than increasing the dose. PMID- 15893554 TI - Modelling the flow [corrected] cytometric data obtained from unperturbed human tumour cell lines: parameter fitting and comparison. AB - In this paper we firstly present three alternative formulations of a mathematical model for human tumour cell lines unperturbed by cancer therapy. The model counts the number density of cells in each phase of the cell cycle over time where cells are differentiated by their DNA content. Data are available from the Auckland Cancer Society Research Centre, Auckland, New Zealand, in the form of DNA histograms or profiles from 11 different human tumour cell lines (i.e. in vitro) unperturbed by cancer therapy. We then apply one (computationally fast) formulation of the model and discover that although in general different combinations of parameter values give rise to very different DNA profiles it is possible that different combinations of parameter values give rise to virtually identical profiles. Experimental estimates of the rate of transition from the G1 phase (growth) to the S-phase (DNA synthesis) enable us to uniquely determine other model parameters of interest that give the least square error between the model and data. We finally apply our model to each of the 11 different cell lines and compare cell cycle phase transit times. Although the DNA histograms of each of the cell lines have similar shapes these cell lines have different combinations of transit times to each other, which could explain why they often react very differently when exposed to anti-cancer therapies during laboratory experiments. An understanding of the in vitro situation may give an insight into why some human cancer patients do not respond to cancer therapy. PMID- 15893555 TI - Modeling physiological resistance in bacterial biofilms. AB - A mathematical model of the action of antimicrobial agents on bacterial biofilms is presented. The model includes the fluid dynamics in and around the biofilm, advective and diffusive transport of two chemical constituents and the mechanism of physiological resistance. Although the mathematical model applies in three dimensions, we present two-dimensional simulations for arbitrary biofilm domains and various dosing strategies. The model allows the prediction of the spatial evolution of bacterial population and chemical constituents as well as different dosing strategies based on the fluid motion. We find that the interaction between the nutrient and the antimicrobial agent can reproduce survival curves which are comparable to other model predictions as well as experimental results. The model predicts that exposing the biofilm to low concentration doses of antimicrobial agent for longer time is more effective than short time dosing with high antimicrobial agent concentration. The effects of flow reversal and the roughness of the fluid/biofilm are also investigated. We find that reversing the flow increases the effectiveness of dosing. In addition, we show that overall survival decreases with increasing surface roughness. PMID- 15893556 TI - Novel moment closure approximations in stochastic epidemics. AB - Moment closure approximations are used to provide analytic approximations to non linear stochastic population models. They often provide insights into model behaviour and help validate simulation results. However, existing closure schemes typically fail in situations where the population distribution is highly skewed or extinctions occur. In this study we address these problems by introducing novel second- and third-order moment closure approximations which we apply to the stochastic SI and SIS epidemic models. In the case of the SI model, which has a highly skewed distribution of infection, we develop a second-order approximation based on the beta-binomial distribution. In addition, a closure approximation based on mixture distribution is developed in order to capture the behaviour of the stochastic SIS model around the threshold between persistence and extinction. This mixture approximation comprises a probability distribution designed to capture the quasi-equilibrium probabilities of the system and a probability mass at 0 which represents the probability of extinction. Two third-order versions of this mixture approximation are considered in which the log-normal and the beta binomial are used to model the quasi-equilibrium distribution. Comparison with simulation results shows: (1) the beta-binomial approximation is flexible in shape and matches the skewness predicted by simulation as shown by the stochastic SI model and (2) mixture approximations are able to predict transient and extinction behaviour as shown by the stochastic SIS model, in marked contrast with existing approaches. We also apply our mixture approximation to approximate a likelihood function and carry out point and interval parameter estimation. PMID- 15893557 TI - Fertility assurance and breeding synchrony. AB - Extrapair fertilizations are an important factor in reproduction of many bird species. It has been suggested that pursuit of extrapair fertilizations provides a selective pressure capable of affecting the "ecological" attributes of some bird species. It is known that in some cases the pursuit of extrapair copulations is motivated by fertility assurance. That is, there are populations in which (i) some males and females cannot produce viable offspring, and (ii) there is no pre copulatory way to choose a compatible mate. In such situations individuals mate for "economic" reasons and attempt to secure reproduction by copulating with multiple partners. Here we formulate and analyze a simple game theoretical model addressing the interplay between environmental constraints and the pursuit of fertility assurance via extrapair copulations in determining breeding synchrony. Our results indicate that breeding synchrony in such cases is determined by the magnitude of the costs (due to environmental constraints) of breeding asynchronously versus the worth of the attendant increase in extrapair opportunities. PMID- 15893558 TI - Duration of an intermittent episode of viremia. AB - HIV-1 infected patients after being treated with potent combinations of antiretroviral drugs for 2-6 months typically reach a state in which virus can no longer be detected within their blood. These patients with undetectable virus occasionally have viral load measurements that are above the limit of detection of current assays. Such measurements are called blips. Here we examine the possibility that such blips represent infrequent measurements taken during a period of time in which there is a transient elevation of virus in the patient's blood, i.e., a so-called transient episode of viremia. By analyzing time series of blips from a large number of patients, we conclude that transient episodes of viremia exist and that on average they extend for a period of about 3 weeks. PMID- 15893559 TI - Analysis of Ebola virus and VLP release using an immunocapture assay. AB - Ebola virus (EBOV), an emerging pathogen, is the causative agent of a rapidly progressive hemorrhagic fever with high mortality rates. There are currently no approved vaccines or treatments available for Ebola hemorrhagic fever. Standard plaque assays are currently the only reliable techniques for enumerating the virus. Effective drug-discovery screening as well as target identification and validation require simple and more rapid detection methods. This report describes the development of a rapid ELISA that measures virus release with high sensitivity. This assay detects both Ebola virus and EBOV-like particles (VLPs) directly from cell-culture supernatants with the VP40 matrix protein serving as antigen. Using this assay, the contribution of the EBOV nucleocapsid (NC) proteins in VLP release was determined. These findings indicate that a combination of NC proteins together with the envelope components is optimal for VLP formation and release, a finding that is important for vaccination with Ebola VLPs. Furthermore, this assay can be used in surrogate models in non biocontainment environment, facilitating both basic research on the mechanism of EBOV assembly and budding as well as drug-discovery research. PMID- 15893560 TI - An inhibition enzyme-linked immunosorbent assay for the detection of antibody to Rift Valley fever virus in humans, domestic and wild ruminants. AB - This paper describes the development and validation of an inhibition ELISA based on gamma-irradiated tissue culture-derived antigen for the detection of antibody to Rift Valley fever virus (RVFV) in humans, domestic and wild ruminants. Validation data sets derived from field-collected sera in Africa (humans=1367, cattle=649, goats=806, sheep=493, buffalo=258, camels=156) were categorized according to the results of a virus neutralisation test. In addition, individual sera from 93 laboratory workers immunized with inactivated RVF vaccine, 136 serial bleeds from eight sheep experimentally infected with wild-type of RVFV, and 200 serial bleeds from 10 sheep vaccinated with the live-attenuated strain of the virus, were used to study the kinetics of RVFV antibody production under controlled conditions. At cut-off values selected at 95% accuracy level by the two-graph receiver operating characteristic analysis the ELISA sensitivity ranged from 99.47% (humans) to 100% (sheep, buffalo, camels). The specificity ranged from 99.29% (sheep) to 100% (camels). Compared to virus neutralisation and haemagglutination-inhibition tests, the ELISA was more sensitive in detection of the earliest immunological responses in experimentally infected and vaccinated sheep. Our results demonstrate that the ELISA format reported here can be used as a safe, robust and highly accurate diagnostic tool in disease-surveillance and control programmes, import/export veterinary certification, and for monitoring of the immune response in vaccinees. PMID- 15893561 TI - A novel accurate amplification created restriction site method for determination of the wild type and the precore mutant hepatitis B virus variants. AB - The most commonly occurring hepatitis B virus (HBV) mutation is the G to A mutation at nucleotide 1896 in the precore region. The aim of this study was to develop a novel accurate amplification created restriction site (ACRS) method for determination of the TGG wild type and the TAG precore mutant HBV variants. Two conserved and consensus specific and diagnostic primers introducing BstXI and XagI cleavage sites were designed in order to determine the G1896 wild type and the A1896 precore mutant HBV variants in all HBV genotypes. The results of the ACRS method were compared with sequencing data. With the ACRS method, three different patterns could be distinguished for the wild type, the precore mutant and mixed infection HBV variants. The results of the ACRS method on 30 HBV isolates revealed the TAG precore mutant in 50% (15/30), the TGG wild type variant in 30% (9/30) and the mixed infection in 20% (6/30). The sequencing data of these samples were in agreement with the ACRS results. The ACRS method is a rapid and cost-effective technique for detecting both the TGG wild type and the TAG HBV precore mutant variants. It can be carried out for follow-up of G1896A precore mutant variant in hepatitis B virus infected subjects at routine molecular diagnostic laboratories. PMID- 15893562 TI - Comparative study of the PrPBSE distribution in brains from BSE field cases using rapid tests. AB - The distribution of PrP(BSE) in the brain of nine confirmed BSE field cases was analyzed using immunohistochemistry and compared to the levels of PrP(BSE) determined by two rapid tests (Prionics-Check WESTERN and Prionics-Check LIA). Each brain was dissected into 16 areas: spinal cord, medulla oblongata, pons, mesencephalon, thalamus, hippocampus, cerebellar vermis, cerebellar medulla, cerebellar hemispheres, occipital cortex, temporal cortex, parietal cortex, striatum, frontal cortex, piriform lobe and olfactory bulbs. The highest levels of PrP(BSE) were detected in the medulla oblongata, spinal cord and pons, and correspondingly both rapid tests showed 100% correlation with the immunohistochemistry with regard to sensitivity and specificity. Some inconsistencies between the levels of PrP(BSE) determined either by immunohistochemistry or by the rapid tests were found in brain areas with medium to low levels of PrP(BSE). These brain areas included the cerebellar hemisphere, olfactory bulb, and the temporal and parietal cortices. A brain PrP(BSE) distribution curve (BPDC) was designed by plotting the PrP(BSE) signals obtained from the two rapid tests versus the anatomical region along the caudal-rostral axis of the brain. Comparison of the BPDC of the nine BSE cases showed that all cases had a similar PrP(BSE) distribution in the brain but with variable intensities, which could be explained by different stages in the progression of the disease. We propose that the BPDC could be used as a tool to differentiate classical cases of BSE from the recently identified atypical BSE cases. PMID- 15893563 TI - Fluorometric cell-ELISA for quantifying rabies infection and heparin inhibition. AB - The purpose of the present study was to implement a fluorometric method for detecting and quantifying viral antigens in human meduloblastoma cells infected by two types of fixed rabies virus (CVS-MB and CVS-BHK) and a street virus using a cell-enzyme linked immunosorbent assay (cell-ELISA) technique; alkaline phosphatase was used as the antibody-marker enzyme and 4-methyl-umbelliferyl phosphate as the fluorogenic substrate. The system was used for detecting up to 1:10,000 viral inoculums, followed by evaluating the effect of heparin on infection. Infected cultures were reliably differentiated from their respective negative controls in both assays allowing data to be analysed statistically. As reported in another study, heparin produces strong inhibition when the CVS-BHK viral strain is used for infection; it has thus been suggested that it binds to the neural cell adhesion molecule and could be blocked by using this drug. This fluorometric method is less time-consuming, has increased reproducibility and useful for quantitation of collected data and can therefore be considered as a useful tool for research. PMID- 15893564 TI - Quantitative real-time PCR for titration of infectious recombinant AAV-2 particles. AB - In this report, we present a fast, reliable and easy to perform method to quantify infectious titers of recombinant AAV-2 (rAAV-2) particles using the LightCycler technology, which is independent from the therapeutic transgene and without the presence of a marker gene. The method is based on the life cycle of AAV-2: after infection of the host cell, the single stranded (ss) AAV-2 genome is converted into a double stranded (ds) form. Following infection with rAAV-2, HeLa cells were lysed and ssDNA of transcriptionally inactive particles were efficiently removed by ssDNA-specific S1 nuclease digestion. The remaining viral dsDNA can be quantified by quantitative real-time PCR (qPCR). For validation of the new method, rAAV-2 preparations were analyzed by two other standard methods for titration of infectious particles in parallel, i.e. the infectious center assay (ICA) as well as flow cytometry using GFP as a marker. Comparing the infectious titers of 40 different AAV-2 fractions assessed by qPCR with the titers determined by FACS analysis a significant correlation (r=0.87, p<0.001) with a mean ratio of the titers assessed by qPCR and FACS of 1.92 (S.D.+/-1.59) was found. Further, the titers of seven rAAV-2 fractions using qPCR and ICA covering 5 log ranges were compared and a significant correlation was found between the results (r=0.80, p<0.001) with a mean ratio of 3.38 (S.D.+/-1.79), respectively. PMID- 15893565 TI - Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA. AB - This report describes the production of several MAbs against N195 protein, a major immunodomain of SARS CoV nucleocapsid protein [He, Q., Chong, K.H., Chang, H.H., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E., Kwang, J., 2004. Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome. Clin. Diagn. Lab. Immunol. 11 (2) 417-422.]. One representative IgG1 monoclonal antibody (MAb), S-A5D5, was selected and characterized. S-A5D5 reacted specifically react with both recombinant and native nucleocapsid protein of SARS CoV. The reactivity of S-A5D5 with purified N195 protein and utilization of the MAb as a detector antibody to develop an antigen capture ELISA was assessed. As little as 37.5 pg of purified N protein and 50 TCID(50) of SARS CoV could be detected by the antigen capture ELISA. Specific binding of the MAb S-A5D5 to both purified N195 and SARS CoV nucleocapsid antigen was effectively inhibited by human SARS positive serum and guinea pig anti-N195 serum. The N protein in N195-spike recombinant baculovirus infected Sf-9 cells could also be identified. N protein was detected in 18 IFA IgM-positive serum samples collected from SARS confirmed patients, but not in nine samples collected from SARS recovery patient. No false positive results were given when 60 samples from healthy individuals were tested, and no cross-reaction occurred when infectious bronchitis virus (IBV), chicken coronavirus, was tested. This monoclonal antibody-based antigen capture ELISA is thus a powerful tool for early diagnosis of SARS CoV infection. PMID- 15893566 TI - Multicenter evaluation of the NucliSens EasyQ HIV-1 v1.1 assay for the quantitative detection of HIV-1 RNA in plasma. AB - The Nuclisens EasyQ HIV-1 v1.1 assay (Biomerieux) is a real-time detection method combined with NASBA technology designed to measure plasma HIV-RNA. Its performance was assessed in 1008 clinical specimens collected from individuals infected with clade B (774) and non-B (234) HIV-1 variants at four European laboratories. The results were compared with those obtained using three other commercial viral load assays: Cobas Amplicor Monitor HIV-1 v1.5 (Roche), Versant HIV-1 RNA assay (Bayer) and Nuclisens HIV-1 QT (Biomerieux). Overall, the linearity, specificity and reproducibility of the EasyQ assay was comparable with that from the other tests. The correlation coefficient (R) between methodologies was 0.85 for Amplicor; 0.87 for Versant; and 0.91 for Nuclisens. The specificity of the assay was 99.4%. Of note, Versant missed 17% of specimens with non-B subtypes which could be detected by EasyQ, while Amplicor provided similar results than EasyQ. HIV-1 group O specimens were only detected by the EasyQ assay. In conclusion, the performance of the EasyQ assay seems to be similar to that of other HIV-1 viral load tests currently on the market, but it is more sensitive than Versant for HIV-1 non-B subtypes and shows a wider dynamic range than Amplicor. Moreover, as it incorporates the advantage of real-time detection procedures, it facilitates high throughput and short turnaround time. PMID- 15893567 TI - Enumeration of bacteriophages in water by different laboratories of the European Union in two interlaboratory comparison studies. AB - As part of a European project on bacteriophages in bathing waters two interlaboratory comparison studies were carried out (May 1997 and March 1998). During these studies phage reference materials as well as naturally polluted standard samples were analysed in 16 European laboratories. Three groups of bacteriophages were tested using standardised methods: somatic coliphages, F specific RNA-phages and phages of Bacteroides fragilis. Many of the participating laboratories applied one or more of the phage methods for the first time, after a one-week training session in a central laboratory. Nevertheless, the values of repeatability (r=1.35-1.38 calculated on log(10)-scale) and reproducibility (R=1.52-2.04 calculated on log(10)-scale) when analysing phage reference materials were close to the theoretical optimum for a Poisson distribution. When analysing the naturally polluted samples more variation in results within and between laboratories was found (r=1.63-2.34; R=3.10-5.72), in comparison with the results obtained with the pure phage reference materials. PMID- 15893568 TI - Utility of recombinant integrin alpha v beta6 as a capture reagent in immunoassays for the diagnosis of foot-and-mouth disease. AB - Recombinant integrin alpha v beta6 was evaluated as a capture ligand in a sandwich ELISA for the detection and serotyping of foot-and-mouth disease (FMD) virus. Our routinely applied method employs seven serotype-specific rabbit polyclonal antibodies as capture ligands and seven serotype-specific guinea pig polyclonal antibodies as detecting reagents. The recombinant integrin bound FMD virus of all seven serotypes but not that of another vesicular disease, swine vesicular disease (SVD). Considerable heterotypic cross-reactions were evident when using the integrin capture ligand in combination with guinea pig detecting antibodies but totally type-specific reactions resulted when serotype-specific monoclonal antibodies (mabs) were used instead of the guinea pig reagents. The specificity of reaction of the integrin capture/mab detector combination was superior to that of our routinely employed rabbit/guinea pig ELISA and offers an improvement for test interpretation. As a universal trapping reagent for all FMD virus serotypes the alpha v beta6 recombinant protein also has the potential for application in other test procedures for viral identification (e.g. pen-side chromatographic strip-tests, biosensors, immunocapture RT-PCR, antigenic characterization procedures and monoclonal antibody profiling of emerging field virus strains) and in antibody detection assays employed for the diagnosis of FMD. PMID- 15893569 TI - Rapid approach to identify an unrecognized viral agent. AB - For epidemic control, rapid identification and characterization of the responsible unknown agent are crucial. To address this critical question, a method was developed for virus discovery based on a flexible nested-PCR subtraction hybridization. As a positive control, we used hepatitis C virus as a hypothetical unrecognized virus and "discover" it in the sample. Using template switching universal long-PCR to produce large quantities of cDNA, our nested-PCR based subtractive hybridization coupled with a single-strand deletion technology removed most of the common cDNA. Following subtraction hybridization, a cDNA library was constructed and displayed by differential reverse dot blot hybridization. This new genomic subtraction hybridization method will be ideally suited to identify rapidly any previously unrecognized viral agent. PMID- 15893570 TI - Real-time RT-PCR differentiation and quantitation of infectious bursal disease virus strains using dual-labeled fluorescent probes. AB - A real-time RT-PCR assay was developed utilizing dual-labeled fluorescent probes binding to VP4 sequence that are specific to the classical (Cl), variant (V) and very virulent (vv) strains of infectious bursal disease virus (IBDV). The assay was highly sensitive and could detect as little as 3 x 10(2) to 3 x 10(3) copies of viral template. Viral genomic copy number could be accurately assayed over a broad range of 7-8 logs of viral genome. The variant sequence-specific probe was found to be highly specific in detecting isolates classified as variant A, D, E, G and GLS-5, and did not react with classical strains. A total of 130 field and experimental variant strain isolates were tested using this assay. The classical sequence-specific probe also demonstrated high sensitivity and specificity, and positively detected a total of 87 STC isolates, both field and experimental isolates, while differentiating between isolates that were variant and classical strains. The very virulent sequence-specific probe detected positively the Holland vvIBDV isolate and did not react with classical or variant strains. Rapid identification of viral strain is a primary concern to poultry flock health programs to ensure administered vaccines will protect against current strains of virus circulating in the flock. The ability to quantify virus concurrently is also of assistance in identifying the progression of disease outbreaks within the flock. PMID- 15893571 TI - The Lim Benyesh-Melnick antiserum pools for serotyping human enterovirus cell culture isolates--still useful, but may fail to identify current strains of echovirus 18. AB - Neutralisation tests are often used as a second line method of serotyping human enterovirus cell culture isolates after failure of indirect immunofluorescence. We reviewed the results of 85 consecutive neutralisation tests performed routinely on presumptive enterovirus isolates using the Lim Benyesh-Melnick antiserum pools after failure of immunofluorescence on most of them, in order to assess the value of these antiserum pools. Confirmation of serotype was determined by partial VP1 genome sequencing. Forty isolates were serotyped successfully and 45 isolates were untypable by the LBM pools, although 16 of the latter would not be expected to be typable by this means. Of eighteen echovirus 18 isolates tested, only two were typable by neutralisation. Of 15 coxsackievirus A16 isolates only nine were typable by neutralisation. Neutralisation with the LBM pools remains a useful second line method of serotyping human enteroviruses despite difficulties with at least two serotypes. PMID- 15893572 TI - Discrimination of lamivudine resistant minor HIV-1 variants by selective real time PCR. AB - A selective real-time PCR (SPCR) method was developed and evaluated for discrimination of resistance mutations in minor human immunodeficiency virus type 1 (HIV-1) populations, using the M184 mutation site as a model system due to its high clinical importance and the low genetic barrier to its development. The method enabled detection of minor viral populations down to 0.1%, and the relative proportions of different quasispecies could be easily displayed using cycle threshold (C(t)) values. An excellent concordance was found when the assay was compared with direct sequencing and cloning results. The impact of mismatch between virus and primer/probe sequences was evaluated, showing that 3' end mutations in the selective downstream primers were very disruptive and that 5' end polymorphisms in the probe area were directly fatal, while mutations in the middle or the 3' end of the probe were less disruptive. These effects were compensated by introducing wobble bases to accommodate the mutations. This sensitive and reliable point-mutation assay, analyzing M184I/V and other important mutations, will be fruitful in gaining new scientific knowledge about the kinetics of resistance mutations in minor viral populations of HIV-1 infected patients at failure of antiretroviral therapy. PMID- 15893573 TI - Large-scale imaging of cortical network activity with calcium indicators. AB - Bulk loading of calcium indicators has provided a unique opportunity to reconstruct the activity of cortical networks with single-cell resolution. Here we describe the detailed methods of bulk loading of AM dyes we developed and have been improving for imaging with a spinning disk confocal microscope. PMID- 15893574 TI - Neural mechanisms underlying the processing of Chinese words: an fMRI study. AB - The present study employed functional magnetic resonance imaging (fMRI) to investigate the neural mechanisms underlying orthographic, phonological and semantic processing of single character Chinese words. Twelve right-handed native Chinese speakers participated in the study. Three fundamental linguistic tasks including orthographic judgment, phonological matching and semantic association task were used. Our results demonstrated robust activation in the left posterior inferior temporal cortex (BA 37) for all three tasks. While the phonological matching task produced left-lateralized activation in the inferior frontal and parietal regions, semantic association task showed considerable bilateral activation in the inferior frontal and occipito-parietal regions. Direct comparison between phonological matching and semantic association task yielded semantic related activation in the anterior portion of the left inferior frontal gyrus (BA 47) and the right inferior frontal region (Broca's homology; BA 45). Behaviorally, there was no difference in response time between phonological matching and semantic association task. Our findings suggested that differential neural pathways were involved in the processing of meaning and sound of single character Chinese words. The present study provided systemic information of the neural substrates underlying the processing of different components of Chinese language. PMID- 15893575 TI - Hyperbaric exposure with high oxygen concentration enhances oxidative capacity of neuromuscular units. AB - The effects of hyperbaric exposure with high oxygen concentration on spinal motoneurons and the skeletal muscle fibers that they innervate were investigated. Five-week-old male rats were exposed to a hyperbaric (1.25 atmospheric pressure) environment with a high oxygen concentration (35.0%) for 6h daily. The number, cell body size, and oxidative enzyme activity of motoneurons innervating the soleus and plantaris muscles were examined after 8 weeks of hyperbaric exposure. In addition, the fiber type distribution, cell size, and oxidative enzyme activity of the slow soleus and fast plantaris muscles were examined. The oxidative enzyme activity of alpha motoneurons innervating the soleus and plantaris muscles increased after hyperbaric exposure, irrespective of their cell body sizes. The percentage of high-oxidative fibers in the soleus and plantaris muscles increased after hyperbaric exposure. The oxidative enzyme activity of all types of fibers in the soleus and plantaris muscles increased after hyperbaric exposure. It is concluded that hyperbaric exposure with high oxygen concentration enhances the oxidative capacity of neuromuscular units. PMID- 15893576 TI - Feed-forward associative learning for volitional movement control. AB - One of the most difficult problems in motor learning is determining the source of a learning signal, sometimes called an error signal. This problem is hidden in the adaptations of simple reflexive movements by attributing its source to sensory organs. The feed-forward associative motor learning theory proposed here attributes the source to the movement system itself. When a subject performs a corrective movement after his primary movement, the proposed neural learning device learns to associate the primary motor command with the corrective motor command by using a place-coding system. In the subsequent trials, the primary movement will involve a correction due to the participation of this mechanism, thus resulting in better performance. The theory assumes three conditions, namely, that a motor center and the learning device share the same place-encoded motor information; the motor center issues a command and a learning signal simultaneously from the same unit; and a learning signal issued with a corrective command has a heterosynaptic interaction with the previous primary command. The cerebellum is a reasonable candidate for the device satisfying these conditions. The reaction time of a corrective movement, usually 100-300 ms, almost satisfies the coincidence condition for long-term depression of the granule-to-Purkinje synapses. As an application, this theory is demonstrated to account for behavioral results regarding saccadic adaptation. PMID- 15893577 TI - Altered food-anticipatory activity rhythm in Cryptochrome-deficient mice. AB - In nocturnal rodents, restricted feeding to daytime (RF) causes feeding associated diurnal locomotor activity that persists for the next 1-2 days when food is withheld. Along with this anticipatory behavior, the expression pattern of clock genes such as mPer1 and mPer2 changes from a nocturnal to diurnal pattern in the liver and cerebral cortex but not in the suprachiasmatic nucleus (SCN). Whether the molecular clockwork, in which mCry1 and mCry2 genes are essential components, is involved in food-anticipatory circadian rhythms is unknown. In this study, we investigated the impact of the absence of mCRY products upon the locomotion pattern induced by RF. RF caused an increase in daytime activity that lasted even for 2 days after food was withheld, in wild type and mCry1-/-mCry2-/- mice. However, RF-induced activity was less stable and appeared more gradually in mutant mice. Similar results were obtained with mice housed under constant darkness or with SCN-lesioned wild-type and mutant mice. Our data reveal that mCry proteins are basically dispensable for food-entrainable oscillation. However, it is also important to note that mCry deficiency affects the stability and development of RF-induced anticipatory locomotor activity. PMID- 15893578 TI - Electrophysiological and morphological properties of identified crossed tecto reticular neurons in the rat superior colliculus. AB - Previously we classified randomly sampled neurons in the intermediate layer (SI) of the rat superior colliculus (SC) into six subclasses according to their firing responses to depolarizing current pulses and five subclasses based on their morphological properties in slice preparations. In the present study, we investigated properties of a major output cell group of the rat SC (PND 17-24), crossed tecto-reticular neurons (cTRNs), which project to the contralateral medial pontine reticular formation. The cTRNs were identified by retrograde labeling with a fluorescent tracer (n=112). We compared their properties with those of presumed interneurons (n=127). We found that a majority of cTRNs were regular spiking neurons with moderate firing frequency (73%) and were multipolar shaped (66%). The cTRNs had larger membrane capacitance, larger soma size and lower input impedance than presumed interneurons. Electrical stimulation of the superficial gray layer induced oligosynaptic EPSPs in the cTRNs. When bicuculline was added to the extracellular solution, the EPSPs were markedly enhanced and bursting spike responses were induced. The bursting responses were suppressed by applying D-2-amino-5-phosphonovalerate. These results suggest that the cTRNs exhibit NMDA receptor-dependent bursting responses to visual inputs. These observations give insights into the neuronal mechanism of generating burst activity in cTRNs, which triggers orienting behaviors. PMID- 15893579 TI - A serotonin 5-HT1A receptor agonist prevents behavioral sensitization to L-DOPA in a rodent model of Parkinson's disease. AB - Marked fluctuation of dopamine concentration in the striatum following long-term L-DOPA administration contributes to the development of L-DOPA-induced motor complications including L-DOPA-induced dyskinesias and wearing-off in patients with Parkinson's disease. We have shown that pretreatment with 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT), a 5-HT1A (5-hydroxytryptamine) receptor agonist, alleviates fluctuation of dopamine levels in the dopamine-denervated striatum of 6-hydroxydopamine-lesioned (hemiparkinsonian) rats after L-DOPA treatment. To determine whether co-administration of 8-OH-DPAT with L-DOPA prevents L-DOPA-induced motor complications, we examined rotation behavior and levels of messenger RNAs coding for dynorphin and glutamic acid decarboxylase in the striatum of 6-hydroxydopamine-lesioned rats treated with L-DOPA alone or L DOPA + 8-OH-DPAT, twice daily, for 2 weeks. Co-administration of 8-OH-DPAT inhibited an increase of rotation behavior to L-DOPA and L-DOPA-induced increases in levels of messenger RNAs coding for dynorphin and glutamic acid decarboxylase in the dopamine-denervated striatum, both of which are established indices of L DOPA-induced motor complications. These results suggest that pharmaceutical products that stimulate 5-HT1A receptors could prove useful in prevention of the development of L-DOPA-induced motor complications in patients with Parkinson's disease. PMID- 15893580 TI - No evidence for interaction between 5-HT2A receptor and serotonin transporter genes in schizophrenia. AB - This study was to aim at investigating the potential interaction for the serotonin receptor gene (5-HTR) 2A and serotonin transporter gene (5-HTTLPR) polymorphisms in the development of schizophrenia, as well as the interaction of the two polymorphisms in relation with symptomatology, family history, age of onset and antipsychotic response. Genomic DNA analysis with polymerase chain reaction (PCR) was used for the genotyping. One hundred and eleven (111) patients with schizophrenia and 172 normal controls participated in the study. We did not find any association between the individual polymorphism and schizophrenia. The significant interaction effect between 5-HTTLPR and 5-HTR2A polymorphisms on the development of schizophrenia as well as on the antipsychotics response, family history, symptomatology and age at onset, was not found. However, subject with 5 HTR2A*TT genotype were found to have lower age of onset, compared to their counterparts (p=0.01). These results suggest that the interaction between 5 HTTLPR and 5-HTR2A polymorphisms may not contribute to susceptibility to schizophrenia as well as some clinical factors such as antipsychotic response, at least in the Korean population. PMID- 15893581 TI - Identification of a functional CRE in the promoter of Fukuyama congenital muscular dystrophy gene fukutin. AB - We isolated a fragment of the fukutin gene promoter from differentiated human NT2 cells using chromatin immunoprecipitation technique with an anti-CREB antibody. This fragment contained a CRE-like sequence and here we describe its functional validation. The results showed that the element was functional in vitro and in vivo and that CREB in neurons was involved in the transcriptional regulation of the fukutin gene. Moreover, its expression in neurons was regulated by cAMP and calcium ions, known triggers of CREB phosphorylation. To our knowledge, this is the first report on the regulation of fukutin gene by transcription factor CREB in response to the signals generated by synaptic activity. The true biological function of fukutin, the gene responsible for Fukuyama-type congenital muscular dystrophy and mental retardation, is at present not known. However, it has been suggested that it might possess glycosyltransferase activity and its intracellular localization within the Golgi structures is consistent with this function. As such, fukutin might play a significant role in post-translational modification of synaptic proteins in neuronal cells. PMID- 15893582 TI - Rat nicastrin gene: cDNA isolation, mRNA variants and expression pattern analysis. AB - Nicastrin is a type 1 transmembrane glycoprotein that interacts with presenilin, Aph-1, and Pen-2 proteins to form a high molecular complex with gamma secretase activity. Then, nicastrin has a central role in presenilin-mediated processing of beta-amyloid precursor protein and in some aspects of Notch/glp-1 signaling in vivo. Here, we isolated a rat nicastrin cDNA and investigated gene expression in embryonic and adult rat tissues. The predicted amino acid sequence is comprised of 708 residues and showed a high degree of identity with other vertebrate orthologs. Besides full-length nicastrin mRNA, we identified an alternative spliced variant lacking the whole exon 3 and predicted to encode a 62-residue long truncated protein. Full-length nicastrin mRNA was observed to be ubiquitously expressed, while the spliced variant was preferentially transcribed in the nervous system, whether in embryonic or adult neural tissues. Studies performed on primary cell cultures demonstrated that the short isoform was expressed in neurons, but not in astrocyte and microglial cells. Further experiments performed to verify the presence of the variant in neuroblastoma culture failed to show any truncated protein. Treatments by cyclohexamide showed the involvement of a quality control-based surveillance mechanism, which selectively degrades the exon 3-skipped isoform. In summary, this is the first report describing a novel skipped isoform of nicastrin which may suggest a new possible control mechanism based on the alternative splicing and nonsense mediated mRNA decay to regulate brain protein expression and provide newer insights into potential implication in Alzheimer's disease. PMID- 15893583 TI - Contralateral cytokine gene induction after peripheral nerve lesions: dependence on the mode of injury and NMDA receptor signaling. AB - There is increasing evidence that unilateral nerve injury evokes contralateral responses, but the underlying mechanisms are largely unknown. In the present investigation, we analyzed cytokine and chemokine gene induction in contralateral, non-lesioned nerves after sciatic nerve crush and chronic constriction injury (CCI) by quantitative reverse transcriptase polymerase chain reaction in mice. After sciatic nerve crush, contralateral changes in cytokine gene expression were restricted to interleukin (IL)-1beta, which showed a monophasic peak at the first postoperative day. Following CCI, contralateral transcripts for IL-1beta, IL-10 and monocyte chemoattractant protein-1 (MCP-1) were significantly increased already at day 1 and upregulation persisted over the next 4 weeks. In contrast, tumor necrosis factor alpha (TNF-alpha) levels remained unchanged. Contralateral gene induction was restricted to the homonymous opposite sciatic nerve, but spared the femoral nerve. NMDA receptor blockade completely abolished contralateral cytokine expression after CCI on the mRNA level. In contralateral dorsal root ganglia, only IL-10 mRNA levels were modified after nerve injury. Sham operation significantly increased the cytokine and chemokine gene expression at the ipsilateral side, but could not mediate contralateral effects. Our study confirms that nerve injury evokes contralateral responses and identifies NMDA-mediated signaling as one underlying mechanism. PMID- 15893584 TI - Neurotrophin levels in postmortem brains of suicide victims and the effects of antemortem diagnosis and psychotropic drugs. AB - Suicide is a major public health problem but the neurobiological factors of risk are poorly understood. Recent studies have mentioned changes in the serotoninergic system and in neuronal plasticity, as well. The present investigation was undertaken to examine whether there is an abnormality in brain derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) proteins in suicide victims. The effect of diagnosis and drug treatments on the neurotrophins was also assessed. Thirty suicide victims (11 F/19 M) and twenty-four (10 F/14 M) drug-free non-suicide subjects, devoid of psychiatric or neurological disease, were examined. Antemortem diagnoses and toxicological analyses had been performed. The ventral prefrontal cortex (PFC), the hippocampus, and the entorhinal cortex were selected. BDNF and NT-3 levels were assayed either with the Western blot or with the ELISA method. Results indicated a significant decrease in BDNF and NT-3 levels in the hippocampus and PFC (only BDNF) but not in the entorhinal cortex, of suicide victims who were drug-free compared with non suicide controls. The decrease was observed in all suicide victims, regardless of diagnosis. In drug-treated suicide victims, neurotrophin levels were not significantly different from non-suicide controls. This study supports a role of BDNF and NT-3 neurotrophin, in the pathophysiology of suicidal behavior. Anatomically, this role may implicate the hippocampus and the PFC but not the entorhinal cortex. The absence of change in BDNF and NT-3 levels of drug-treated suicide victims suggests that both neurotrophins are mediators of psychotropic drugs. A better understanding of the neurobiology of suicide could help detect populations at risk. PMID- 15893585 TI - Localization of kainate receptor subunit GluR5-immunoreactive cells in the rat hypothalamus. AB - Glutamate is the major excitatory neurotransmitter in the hypothalamus, which exerts its effects by activating ion channel-forming (ionotropic) or G-protein coupled (metabotropic) receptors. Kainate-preferring glutamate receptor subunits (GluR5, GluR6, GluR7, KA1, and KA2) form one of the three ionotropic receptor families. In the present study, we analyzed the distribution of GluR5 subunit protein in the rat hypothalamus with immunohistochemistry. GluR5 immunoreactivity was observed in perikarya and processes of many hypothalamic cells some of which, based upon their morphological differentiation by size and structure, appeared to be neurons and others glial cells. Analyses revealed that higher number of glial cells were GluR5 positive when compared to the moderate number of GluR5-labeled neurons in the anteroventral periventricular nucleus. Numerous GluR5-expressing neurons and similar number of glia were detected in the suprachiasmatic nucleus. In the arcuate nucleus more glial cells were identified with GluR5 immunoreactivity than the number of labeled neurons. Scattered GluR5-positive cells were present in the periventricular nucleus. Specific immunostaining was not seen in the ventromedial nucleus or dorsomedial nucleus. In conclusion, it is suggested that the GluR5 subunits participate in the glutamatergic regulation of several neuroendocrine systems, such as the tubero-infundibular systems as well as in the control of circadian output through neuron-to-neuron and/or neuron-to glia interactions. PMID- 15893586 TI - The regional and cellular expression profile of the melatonin receptor MT1 in the central dopaminergic system. AB - The physiological effects of pineal melatonin are primarily mediated by melatonin receptors located in the brain and periphery. Even though there are a number of studies demonstrating the regulatory role of melatonin in the development of dopaminergic behaviors, such as psychostimulant-induced diurnal locomotor sensitization or drug seeking, little is known about the contribution of melatonin receptors (i.e., MT1) to this role. Therefore, as a first step in understanding the functional role of melatonin receptors in dopaminergic behaviors, we focused on determining the expression pattern of MT1 receptors in the dopaminergic system of the human and rodent brain. Regional (e.g., nucleus accumbens shell) and cellular (e.g., tyrosine hydroxylase immunopositive cells) expression of MT1 mRNA was characterized by applying the immuno-laser capture microdissection (immuno-LCM) technique coupled with nested RT-PCR. Moreover, employing quantitative Western immunoblotting and RT-PCR, we found that the mouse MT1 receptor expression presents diurnal variations (i.e., low mRNA and high protein levels at night, ZT21). The dopaminergic system-based presence of MT1 receptor proteins was not limited to rodents; we found these receptors in postmortem human brain as well. Further research is needed to understand the regional/cellular functional role of melatonin receptors in the regulation of dopaminergic behaviors, using models such as melatonin receptor knockout mice. PMID- 15893587 TI - Purification and mass spectrometric analysis of the delta opioid receptor. AB - A mouse delta opioid receptor was engineered to contain a FLAG epitope at the amino-terminus and a hexahistidine tag at the carboxyl terminus to facilitate purification. Selection of transfected human embryonic kidney (HEK) 293 cells yielded a cell line that expressed the receptor with a B(max) of 10.5 pmol/mg protein. [3H]Bremazocine exhibited high affinity binding to the epitope-tagged delta opioid receptor with a K(D) of 1.4 nM. The agonists DADL, morphine, and DAMGO competitively inhibited bremazocine binding to the tagged delta receptor with K(I)'s of 0.9, 370, and 620 nM, respectively. Chronic treatment of cells expressing the epitope-tagged delta receptor with DADL resulted in downregulation of the receptor, indicating that the tagged receptor retained the capacity to mediate signal transduction. The delta receptor was solubilized from HEK 293 cell membranes with n-dodecyl-beta-d-maltoside in an active form that maintained high affinity bremazocine binding. Sequential use of Sephacryl S300 gel filtration chromatography, wheat germ agglutinin (WGA)-agarose chromatography, immobilized metal affinity chromatography, immunoaffinity chromatography, and SDS/PAGE permitted purification of the receptor. The purified delta opioid receptor was a glycoprotein that migrated on SDS/PAGE with an apparent molecular mass of 65 kDa. MALDI-TOF mass spectrometry was used to identify and characterize peptides derived from the delta opioid receptor following in-gel digestion with trypsin, and precursor-derived ms/ms confirmed the identity of peptides derived from enzymatic digestion of the delta opioid receptor. PMID- 15893588 TI - Early changes in gene expression in the dorsal root ganglia after transection of the sciatic nerve; effects of amphiregulin and PAI-1 on regeneration. AB - To characterize the gene activity that may be required for neuronal survival and regeneration, we used the Affymetrix GeneChip Mu74A to screen 12000 genes and expressed sequence tag (EST) mRNA from L4 and L5 mouse dorsal root ganglia (DRG) 12 h and 24 h after sciatic nerve transection. At 12 h, we found 17 upregulated transcripts, and at 24 h, 49 that met our criteria of a significant 2-fold increase in expression. The alterations included a total of eight transcription factors and several genes associated with TGF-beta- and IL-6-mediated signaling. Two of the changes, amphiregulin and plasminogen activator inhibitor-1 (PAI-1), were confirmed by real-time quantitative PCR (QPCR). Addition of amphiregulin (20 ng/ml) to organ-cultured DRG stimulated axonal outgrowth while PAI-1 (20 nM) inhibited migration of Schwann cells from the ganglia. PMID- 15893589 TI - Up-regulation of gamma-glutamyl transpeptidase (GGT) activity in growth perturbed C6 astrocytes. AB - Activity of gamma-glutamyl transpeptidase (GGT) was studied in astrocyte-like C6 glial cells modulated in growth and maturation by different concentration of serum and dibutyryl cyclic AMP (Db-cAMP) supplement in culture medium. After reduction of serum concentration from 10% to 0.1%, the number of GGT positive cells determined histochemically increased 3.1 times and the GGT activity/mg protein in whole cell lysates was 5.1 times higher. In cultures with 0.1% serum + Db-cAMP, the histochemically and biochemically assayed GGT activity exceeded 5.1 and 7.9 times the values measured in control 10% serum cultures, respectively. The up-regulation of GGT was accompanied by an inhibition of proliferation, enhanced differentiation and hypertrophy of cells. In addition, the process of metabolic perturbation and/or cellular stress was revealed in these cultures by the (i) growth-support release followed by shrinkage and death of a small number of cells and (ii) higher oxidation of 2'7'dichlorofluorescein diacetate to its fluorescent form in the adherent/viable cells. The observed up-regulation of GGT is considered to primarily reflect increased metabolism of glutathione and/or the maintenance of the redox potential in cells stressed by sub-optimal concentration of serum and Db-cAMP supplement. The concomitant cellular hypertrophy and differentiation and their relationship to increased activity of GGT await further investigation. The study suggests that up-regulation of GGT can contribute to adaptation of astrocytic cells to metabolic and/or oxidative perturbances occurring under various pathological conditions, including radiation- and drug induced toxicity. PMID- 15893590 TI - Mutational effects at the tetramerization site of nonerythroid alpha spectrin. AB - Spectrin, a prominent cytoskeletal protein, exerts its fundamental role in cellular function by forming a sub-membrane filamentous network. An essential aspect of spectrin network formation is the tetramerization of spectrin alphabeta heterodimers. We used laboratory methods, the yeast two-hybrid system and random mutagenesis, to investigate, for the first time, effects of amino acid mutations on tetramerization of nonerythroid (brain) spectrin (fodrin). Based on high sequence homology with erythroid spectrin, we assume the putative tetramerization region of nonerythroid alpha-spectrin at the N-terminal region. We introduced mutations in the region consisting of residues 1-45 and studied mutational effects on spectrin alphabeta association to form tetramers. We detected single, double, and triple mutations involving 24 residues in this region. These amino acid mutations of nonerythroid alpha-spectrin exhibit full, partial, or no effect on the association with nonerythroid beta-spectrin. Single amino acid mutations in the region of residues 1-9 (D2Y, G5V, V6D, and V8M) did not affect the association. However, seven single mutations (I15F, I15N, R18G, V22D, R25P, Y26N, and R28P) affected the alphabeta association. These mutations were clustered in the region predicted by sequence alignment to be crucial in nonerythroid alpha spectrin for tetramerization, a region that spanned residues 12-36, corresponding to the partial domain Helix C' (residues 21-45) in erythroid alpha-spectrin. In addition, two other mutations, one upstream and one downstream of this region at positions 10 (E10D) and 37 (R37P), also affected the alphabeta association. Our results implied nonerythroid alpha-spectrin partial domain helix may be longer than Helix C' (residues 21-45 and a total of 25 residues) in erythroid alpha spectrin and spanned at least residues 10-37. It is interesting to note that seven out of these nine single mutations (I15F, I15N, R18G, V22D, R25P, Y26N, R37P) were at the a, d, e or g heptad positions based on sequence alignment with erythroid alpha-spectrin. Four of the mutated residues (I15, R18, V22, R25) are conserved in both erythroid and nonerythroid spectrin. These positions were previously identified as hot spots in erythroid alpha-spectrin that lead to severe hematological symptoms. This study clearly demonstrated that single mutation in a region predicted to be critical functionally in nonerythroid alpha spectrin indeed leads to functional abnormalities and may lead to neurological disorders. PMID- 15893591 TI - TRPM8 protein localization in trigeminal ganglion and taste papillae. AB - TRPM8 is a TRP family cation channel which can be activated by cold stimuli or l menthol. However, TRPM8 protein localization of nerve terminals in sensory organs remains unknown. Here we generated an antibody against TRPM8 and analyzed TRPM8 protein localization in trigeminal ganglia (TG) and in sensory nerve fibers in the tongue. TRPM8 immunoreactivity was detected in a subset of neurons with a small diameter in TG and in nerve fibers in the tongue. TRPM8-immunoreactive nerve fibers were rich in fungiform papillae, but sparse in foliate and circumvallate papillae. The TRPM8-immunoreactive nerve fibers reached the outer epithelial layer in each papilla, while no TRPM8-immunoreactive nerve fibers penetrated into taste buds. Double labeling analysis revealed that TRPM8 immunoreactivity was co-expressed with a part of TRPV1 or CGRP-immunoreactive neurons in TG. However, TRPM8 immunoreactivity was not observed in TRPV1- or CGRP positive nerve fibers in fungiform, foliate, and circumvallate papillae. These results suggest that TRPM8 protein is present in sensory lingual nerve fibers mainly projected from TG and might work as cold and l-menthol receptors on tongue. PMID- 15893592 TI - Robustness of gene expression profiling in glioma specimen samplings and derived cell lines. AB - One of the most promising applications of microarrays is class distinction through gene expression profiling as a diagnostic tool. However, as there is apparent spatial heterogeneity in the morphology of cancer cells within a tumor, it is unclear if tumor sampling can be applied and yield consistent signals. In this report, we examined six brain tumors, four glioblastoma, and two oligodendroglioma biopsies. The six brain tumor tissues from two distinct different classes were dissected in four distinct areas and gene expression was profiled using microarrays. We used hierarchical clustering to compare the variability of gene expression profiles between spatially distinct biopsies of the same tumor as compared to the variability between tumors of the same histologic group. We conclude that, in general, repeat spatially distinct samples are not needed for microarray experiments and the gene expression signatures are robust across the tumor. Predominantly, variation was much greater between samples from different patients than from the multiple samplings of given tumor. Further, we compared biopsy expression profiles to the cell lines derived from those tissues. In general, the tumor cell lines vary greatly from the parental tissues and cluster more strongly with each other than the parental tissue. We select and examine the set of genes altered in expression to allow adaptation to cell culture. PMID- 15893593 TI - Magnesium blocks the loss of protein kinase C, leads to a transient translocation of PKC(alpha) and PKC(epsilon), and improves recovery after anoxia in rat hippocampal slices. AB - Magnesium is a potent neuroprotective agent against damage to synaptic transmission during cerebral anoxia and reoxygenation. We investigated the mechanisms of anoxic transmission damage and magnesium neuroprotection by examining the response of PKC isoforms to an anoxic insult in the rat hippocampal slice model. A 2-min anoxic period, which resulted in almost complete recovery of synaptic function, did not result in PKC downregulation. In contrast, inducing long-term damage with 10-min anoxia resulted in the downregulation of the conventional PKCs betaI, betaII and gamma immediately after the insult and after 1-h reoxygenation. There was additional loss of PKC(alpha) and PKC(epsilon) after 1-h reoxygenation. Magnesium treatment improved the recovery of synaptic transmission, blocked the loss of PKC and resulted in a transient translocation of PKC(alpha) and PKC(epsilon) to the membrane fraction. Selective downregulation of cPKCs and PKC(epsilon) correlated with permanent damage to synaptic transmission while translocation of PKC(alpha) and PKC(epsilon) correlated with preservation of synaptic function. The mechanisms of magnesium neuroprotection may include altering the PKC response to an anoxic insult. PMID- 15893594 TI - Genetic variants of angiotensin converting enzyme and methylenetetrahydrofolate reductase may act in combination to increase migraine susceptibility. AB - Migraine, with and without aura (MA and MO), is a prevalent and complex neurovascular disorder that is likely to be influenced by multiple genes some of which may be capable of causing vascular changes leading to disease onset. This study was conducted to determine whether the ACE I/D gene variant is involved in migraine risk and whether this variant might act in combination with the previously implicated MTHFR C677T genetic variant in 270 migraine cases and 270 matched controls. Statistical analysis of the ACE I/D variant indicated no significant difference in allele or genotype frequencies (P > 0.05). However, grouping of genotypes showed a modest, yet significant, over-representation of the DD/ID genotype in the migraine group (88%) compared to controls (81%) (OR of 1.64, 95% CI: 1.00-2.69, P = 0.048). Multivariate analysis, including genotype data for the MTHFR C677T, provided evidence that the MTHFR (TT) and ACE (ID/DD) genotypes act in combination to increase migraine susceptibility (OR = 2.18, 95% CI: 1.15-4.16, P = 0.018). This effect was greatest for the MA subtype where the genotype combination corresponded to an OR of 2.89 (95% CI:1.47-5.72, P = 0.002). In Caucasians, the ACE D allele confers a weak independent risk to migraine susceptibility and also appears to act in combination with the C677T variant in the MTHFR gene to confer a stronger influence on the disease. PMID- 15893595 TI - Expression of the protein inhibitor of nitric oxide synthase in the adult rat retina before and after optic nerve lesion. AB - The molecular messenger nitric oxide (NO) not only serves a number of physiologic functions, but is also involved in the pathophysiology of neurodegeneration. It is produced by the nitric oxide synthase (NOS) isoenzymes. One of the many players regulating NOS activity is the Protein Inhibitor of NOS, PIN. To gain further insight into the mechanisms of NOS regulation and NO-mediated cell death after nerve trauma, we examined PIN expression in a standard model of lesion induced neurodegeneration, the rat optic nerve transsection model. In both the axotomized retinae and the control retinae PIN expression was predominantly observed in the retinal ganglion cell layer. Optic nerve lesion did neither change the amount of PIN mRNA, as determined by in situ hybridization and real time RT-PCR, nor did it change the amount of PIN as determined by immunohistochemistry and Western blot analysis. These results suggest that in our model, NOS activity is not regulated by altered PIN levels, which contributes to our understanding of apoptotic mechanisms in injured neurons. PMID- 15893596 TI - In situ hybridization evidence for the coexistence of ASIC and TRPV1 within rat single sensory neurons. AB - The activation of nociceptors by protons plays a crucial role in the initiation and maintenance of acidosis-linked pain. Acid-sensing ion channel (ASIC) and transient receptor potential/vanilloid receptor subtype-1 (TRPV1) encode proton activated cation channels expressed by nociceptors and the opening of these channels results in nociceptor excitation. Histological relations among ASIC clones and the colocalization of each ASIC subunit and TRPV1 within single sensory neurons were examined on serial sections of rat dorsal root ganglia (DRG) using in situ hybridization histochemistry. ASIC1a transcripts were expressed in 20-25% of the DRG neurons, and most of the neurons had small (<30 microm) diameter cell bodies. ASIC1b transcripts and ASIC3 transcripts were expressed in approximately 10% and 30-35% of the DRG neurons, respectively, and the greater part of each population was located in small-to-medium (30-50 microm)-diameter cells. The ASIC1a transcripts and ASIC1b transcripts were basically localized in the distinct populations of the DRG neurons, while approximately 20% of the ASIC1a-positive neurons and approximately 10% of the ASIC1b-positive neurons expressed ASIC3 transcripts. TRPV1 transcripts were expressed in 35-40% of the DRG neurons, and most of the TRPV1-positive neurons had small-diameter cell bodies. Intense expression signals for ASIC1a transcripts were detected in 40-45% of the TRPV1-positive neurons. Neurons expressing both ASIC1b and TRPV1 transcripts were barely detected in the DRG. Approximately 30% of the TRPV1 positive neurons expressed ASIC3 transcripts, and the double-labeled neurons were comprised of both small-diameter and medium-diameter cells. Approximately 13% of the TRPV1-positive neurons expressed both ASIC1a and ASIC3 transcripts. PMID- 15893597 TI - Differential localization of MAPK-activated protein kinases RSK1 and MSK1 in mouse brain. AB - RSK1 and MSK1 are closely related members of the MAP kinase-activated kinase family and are direct substrates and effectors of the well-studied mitogen activated protein kinases. Although extensively characterized at the biochemical level, little is known about the localization of these protein kinases in the brain. We utilized immunohistochemistry to determine the cellular and subcellular localization of RSK1 and MSK1 in the adult mouse brain. RSK1 is expressed at highest levels in cerebellum, especially in granule neurons and within neuropil of the molecular layer. RSK1 is also expressed in microglia throughout the brain. In a focal trauma model, RSK1 immunoreactivity is increased in activated microglia. RSK1 expression is also prominent in many large pyramidal neurons throughout the brain. At the subcellular level, RSK1 is highly concentrated in the golgi apparatus of both neurons and astroglia. In contrast, MSK1 is expressed at highest levels in striatal and olfactory tubercle neurons and to a lesser degree in cerebellar Purkinje cells. MSK1 is also expressed in a subset of astroglia. At the subcellular level, MSK1 is confined to the nucleus of all expressing cell types. The differential cellular and subcellular localizations of RSK1 and MSK1 suggest divergent functional roles in the brain, with RSK1 poised to regulate membrane trafficking or membrane-localized signaling, and MSK1 involved in modification of nuclear histones and transcription factors. PMID- 15893598 TI - In situ hybridization of dihydroxyacetone phosphate acyltransferase, the regulating enzyme involved in plasmalogen biosynthesis. AB - In situ hybridization can be carried out using different methods. The experimenter has to choose various parameters: the type of tissue fixation, the time of incubation, and the duration of the exposure time. All these parameters are determinant for the sensitivity and the resolution of this technique. This publication of technical aspects described different experiments performed for in situ hybridization on liver tissue. We may conclude on the parameters to optimize each step of the hybridization procedure. Moreover, this technique could be transposed to the brain and applied to little structures with a light expression of DHAP-AT. PMID- 15893599 TI - Opposite effects of sleep rebound on orexin OX1 and OX2 receptor expression in rat brain. AB - Orexins (hypocretins) have been implicated in the regulation of the normal sleep wake cycle, in sensorimotor programming, and in other homeostatic and neuroregulatory processes. The present study examined the effects of sleep deprivation (SD) and sleep recovery on the expression of orexin 1 receptors (OX1R) and orexin 2 receptors (OX2R) throughout the brain. Rats were sacrificed either immediately after 96 h of sleep deprivation (SD group) or after SD followed by 24 h of sleep recovery (Rebound group). Prepro-orexin mRNA showed a non-significant increase in the SD group relative to controls, but a pronounced and significant increase in the Rebound group (+88%, P < 0.007). Similarly, sleep deprivation produced no effect on OX1R or OX2R mRNA levels. However, in the Rebound group, OX1R mRNA levels increased significantly, compared to either control or SD groups, in 37 of 92 brain regions analyzed, with particularly strong effects in the amygdala and hypothalamus. Changes in OX2R mRNA levels were also seen only in the sleep Rebound group, but they were fewer in number (10 out of 86 regions), were in the direction of decreased rather than increased expression, and were predominantly confined to cerebral cortical areas. These observations indicate that some factor associated with sleep recovery, possibly the compensatory increase in REM sleep, has strong effects on the orexin system at the mRNA level. They further indicate that,pOX1 and OX2 receptors are affected in opposite way and that the former are more vulnerable to these effects than the latter. PMID- 15893600 TI - Intercellular trafficking activity of herpes simplex virus US11 gene product in the mouse brain. AB - The US11 gene product of herpes simplex virus is an RNA-binding protein with a C terminal arginine-X-proline (RXP) repeating motif. We found that the RXP repeat mediates intercellular trafficking activity and accumulation in neuronal nuclei following in vivo transfection with the US11 gene, direct injection of the purified RXP-repeat fusion protein, or infection with herpes simplex virus. We discuss a possible therapeutic application of the US11 protein RXP repeat as a tactic against neurodegenerative disorders of the central nervous system. PMID- 15893601 TI - From synapse to gene product: prolonged expression of c-fos induced by a single microinjection of carbachol in the pontomesencephalic tegmentum. AB - It is not known how the brain modifies its regulatory systems in response to the application of a drug, especially over the long term of weeks and months. We have developed a model system approach to this question by manipulating cholinergic cell groups of the laterodorsal and pedunculopontine tegmental (LDT/PPT) nuclei in the pontomesencephalic tegmentum (PMT), which are known to be actively involved in the timing and quantity of rapid eye movement (REM) sleep. In a freely moving feline model, a single microinjection of the cholinergic agonist carbachol conjugated to a latex nanosphere delivery system into the caudolateral PMT elicits a long-term enhancement of one distinguishing phasic event of REM sleep, ponto-geniculo-occipital (PGO) waves, lasting 5 days but without any significant change in REM sleep or other behavioral state. Here, we test the hypothesis that cholinergic activation within the caudolateral PMT alters the postsynaptic excitability of the PGO network, stimulating the prolonged expression of c-fos that underlies this long-term PGO enhancement (LTPE) effect. Using quantitative Fos immunohistochemistry, we found that the number of Fos immunoreactive (Fos-IR) neurons surrounding the caudolateral PMT injection site decreased sharply by postcarbachol day 03, while the number of Fos-IR neurons in the more rostral LDT/PPT increased >30-fold and remained at a high level following the course of LTPE. These results demonstrate a sustained c-fos expression in response to pharmacological stimulation of the brain and suggest that carbachol's acute effects induce LTPE via cholinergic receptors, with subsequent transsynaptic activation of the LDT/PPT maintaining the LTPE effect. PMID- 15893602 TI - NF-(kappa)B mediates amyloid beta peptide-stimulated activity of the human apolipoprotein E gene promoter in human astroglial cells. AB - The apolipoprotein E gene (APOE) plays an important role in the pathogenesis of Alzheimer's disease (AD), and amyloid plaque comprised mostly of the amyloid-beta peptide (A(beta)) is one of the major hallmarks of AD. However, the relationship between these two important molecules is poorly understood. We examined how A(beta) treatment affects APOE expression in cultured cells and tested the role of the transcription factor NF-(kappa)B in APOE gene regulation. To delineate NF (kappa)B's role, we have characterized a 1098 nucleotide (nt) segment containing the 5'-flanking region of the human APOE gene (-1054/+44, +1 transcription start site). Sequence analysis of this region suggests the presence of two potential NF (kappa)B elements. To demonstrate promoter activity, the region was cloned upstream of a promoterless luciferase (reporter) gene. This segment was able to drive expression of luciferase in transient transfections of human fetal glial cells. Promoter activity was stimulated twofold by A(beta)(1-40) (25 microM, 24 h) treatment. Pretreatment with double-stranded DNA decoy oligonucleotides against NF-(kappa)B (2 microM) reduced A(beta) stimulation. Deletion and mutagenetic analyses demonstrated that the distal NF-(kappa)B element was functional and showed a strong DNA-protein complex band in gel shift analysis, similar to that from control NF-(kappa)B consensus element. An anti-inflammatory and anti-NF-(kappa)B drug, sodium salicylate, significantly blocked A(beta) induced APOE promoter function. Our data provide evidence that upregulation of APOE by A(beta) in astroglial cells is mediated by an NF-(kappa)B-element present in the 5'-flanking region of the APOE gene. PMID- 15893603 TI - Cloning, characterization and neuronal expression profiles of tumor endothelial marker 7 in the rat brain. AB - Tumor endothelial marker7 (TEM7) is a putative transmembrane protein that is highly expressed in the tumor endothelium. In the present study, the expression profile of TEM7 was investigated in TEM7-transfected human embryonic kidney (HEK) 293 cells and the rat brain. The extracellular secretion of the recombinant N terminal ectodomain of TEM7, not full-length TEM7, was observed in the transiently transfected HEK 293 cells. The full-length TEM7 was found inside and membrane part of cells as demonstrated by confocal microscopy. In situ hybridization study revealed that TEM7 mRNA expressions were localized to specific neuronal areas, such as cerebellar Purkinje cells, the layer IV and V of cerebral cortex, hippocampal pyramidal cells and hypothalamic magnocellular nuclei. Immunohistochemical investigation of TEM expression with specific antibodies against TEM7 further supported the spatial expression patterns of TEM7 mRNA. The temporal expression of TEM7 mRNA in the cerebellar Purkinje cells demonstrated a postnatal developmental regulation of TEM7 expression. Our results indicate that TEM7 plays a role as a transmembrane receptor in some neuronal populations of the vertebrate brains. PMID- 15893604 TI - The circadian clock-containing photoreceptor cells in Xenopus laevis express several isoforms of casein kinase I. AB - The frog (Xenopus laevis) retina has been an important model for the analysis of retinal circadian rhythms. In this paper, several isoforms of X. laevis casein kinase I (CKI) were analyzed to address whether they are involved in the phosphorylation and degradation of period protein (PER), as they are in the circadian oscillators of other species. cDNAs encoding two splice variants of CKI(delta) (a full-length form and deletion isoform, which is missing an exon that encodes a putative nuclear localization signal and two evolutionarily conserved protein kinase domains) were isolated and analyzed, together with a previously isolated CKI(epsilon) isoform. Both CKI(delta) and CKI(epsilon) were shown to be constitutively expressed in the photoreceptors of the retina, where a circadian clock has been localized. Both the full-length CKI(delta) and CKI(epsilon) were shown to have kinase activity in vitro, and the full-length CKI(delta) phosphorylated and degraded Drosophila PER when expressed in Drosophila S2 cells. The expression and biochemical characteristics of these CKIs are consistent with an evolutionarily conserved role for CKI in the Xenopus retinal clock. The CKI(delta) deletion isoform did not exhibit kinase activity and did not trigger degradation of PER. Subcellular localization of both CKI(delta) isoforms was cytoplasmic in several cell culture lines, but the full length CKI(delta) , and not the deletion CKI(delta) isoform, was localized to both the nucleus and the cytoplasm in Drosophila S2 cells. These results indicate that the sequences missing in the deletion CKI(delta) isoform are important for the nuclear localization and kinase activity of the full-length isoform and that one or both of these features are necessary for degradation of Drosophila PER. PMID- 15893605 TI - Anti-angiogenic activity of the mutant Dutch A(beta) peptide on human brain microvascular endothelial cells. AB - Cerebral amyloid angiopathy is a common pathological feature of patients with Alzheimer's disease (AD) and it is also the hallmark of individuals with a rare autosomal dominant disorder known as hereditary cerebral hemorrhage with amyloidosis-Dutch type. We have shown previously that wild type A(beta) peptides are anti-angiogenic both in vitro and in vivo and could contribute to the compromised cerebrovascular architecture observed in AD. In the present study, we investigated the potential anti-angiogenic activity of the Dutch A(beta)(1-40) (E22Q) peptide. We show that compared to wild type A(beta), freshly solubilized Dutch A(beta) peptide more potently inhibits the formation of capillary structures induced by plating human brain microvascular endothelial cells onto a reconstituted basement membrane. Aggregated/fibrillar preparations of wild type A(beta) and Dutch A(beta) do not appear to be anti-angiogenic in this assay. The stronger anti-angiogenic activity of the Dutch A(beta) compared to wild type A(beta) appears to be related to the increased formation of low molecular weight A(beta) oligomers in the culture medium surrounding human brain microvascular endothelial cells. Using oligonucleotide microarray analysis of human brain microvascular endothelial cells, followed by a genome-scale computational analysis with the Ingenuity Pathways Knowledge Base, networks of genes affected by an anti-angiogenic dose of Dutch A(beta) were identified. This analysis highlights that several biological networks involved in angiogenesis, tumorigenesis, atherosclerosis, cellular migration and proliferation are disrupted in human brain microvascular endothelial cells exposed to Dutch A(beta). Altogether, these data provide new molecular clues regarding the pathological activity of Dutch A(beta) peptide in the cerebrovasculature. PMID- 15893606 TI - Gene expression profiling during the embryonic development of mouse brain using an oligonucleotide-based microarray system. AB - We analyzed gene expression profiles in embryonic day 12, 15, 18 and postnatal day 0 mouse brains by utilizing a GeneChip microarray. Significant differential expression was observed in 1413 of 12,422 (11.4%) represented on the chip. Then, 397 genes known to be related to neural development and functions were selected and analyzed in more detail. Clustering of the differentially expressed genes in terms of gene function and their temporal expression patterns indicated an aspect of the genetic foundation that underlies cellular events. Moreover, we identified a novel gene that encodes a putative protein kinase, Ebr kinase, which is differentially expressed in the developing brain. PMID- 15893607 TI - Molecular cloning, expression, purification, and characterization of shorter forms of human glutamic decarboxylase 67 in an E. coli expression system. AB - Previously, we reported the presence of truncated form of human brain l-glutamic decarboxylase 65 (tGAD65) in vivo as well as in vitro and found that tGAD65 was more active than the full-length GAD65 (Wei et al., J. Biomed. Sci., 10: 617-624, 2003). Here, we report the presence of two shorter forms of hGAD67, namely, hGAD67 (Delta1-70) and hGAD(67) (Delta1-90), referring to a deletion of 1-70 and 1-90 amino acids from the N-terminal, respectively. The molecular masses of hGAD67 (Delta1-70) and hGAD67 (Delta1-90) were found to be 59 kDa and 57 kDa, respectively. Both shorter forms were cloned, expressed, and characterized. In contrast to hGAD65, the shorter forms of hGAD67 were much less active than the full-length due to decrease in affinity of PLP towards the shorter enzymes. Both the full-length and one of the shorter forms of GAD67 were detected in porcine brain extract. Furthermore, the full-length GAD67 could be converted to both shorter forms by crude brain extract, suggesting that an endogenous protease may be present in the brain, which is responsible for the conversion. The cleavage of GAD67 seems to be Ca+(2)-dependent. The model for the conversion of GAD from full length GAD to shorter forms of GAD and its physiological implications was proposed. PMID- 15893608 TI - Prepubertal chronic ethanol administration alters TTF-1 and Oct-2 expression in the hypothalamus of female rats. AB - We found that prolonged administration of ethanol (3 g/kg i.p. at 08:00, once per day) to young female rats starting on postnatal day 24 caused delayed puberty. We further found that prolonged ethanol administration changed the typical hypothalamic expression patterns of TTF-1 and Oct-2 and reduced GnRH mRNA expression. We suggest that these changes may cause the ethanol-induced disturbances in the regulation of GnRH in the hypothalamus and may be responsible for the ethanol-induced reduction in GnRH and LH associated with delayed puberty. PMID- 15893609 TI - A role of IL-1 in MPTP-induced changes in striatal dopaminergic and serotoninergic transporter binding: clues from interleukin-1 type I receptor deficient mice. AB - In mice, the MPTP-induced striatal dopaminergic denervation is followed by a spontaneous partial DAT recovery and by serotoninergic hyperinnervation. We show that IL-1RI-deficient mice have a higher DAT decrease in the ventromedial striatum after MPTP and a higher basal serotoninergic innervation of the whole striatum. These data point to a possible role of IL-1RI in the early MPTP-induced structural or functional remodeling of the nigrostriatal dopamine system. PMID- 15893610 TI - delta-Opioid agonist induced regulation of E2F1 DNA binding activity in NG108-15 cells. AB - Activation of opioid receptors have been implicated in the modulation of cell proliferation and the E2F family of transcription factors may play a role in opioid inhibition of DNA synthesis. Gel shift assays and Western blotting of nuclear extracts from NG108-15 cells revealed increased E2F1 DNA binding activity and higher levels of E2F1 following activation of delta-opioid receptors. It is suggested that DADLE-induced regulation of E2F DNA binding activity involves ERKs. PMID- 15893611 TI - Upregulation of mRNAs coding for AMPA and NMDA receptor subunits and metabotropic glutamate receptors in the dorsal horn of the spinal cord in a rat model of diabetes mellitus. AB - Recent studies suggest that glutamate plays a pivotal role in the processing of sensory information in the spinal cords of patients with diabetic neuropathy. However, the specific glutamate receptors that that are involved have yet to be determined. We therefore conducted a study to characterize the expression of messenger RNAs (mRNAs) coding for subunits of alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA) receptors and N-methyl-d-aspartate (NMDA) receptors and for metabotropic glutamate receptors (mGluRs) in the dorsal horn of the lumbar segment of the spinal cord in a rat model (streptozotocin [STZ] induced) of diabetic neuropathy. The levels of mRNAs coding for AMPA receptor subunits, GluR1, GluR2, and GluR3, were significantly increased in all layers (laminae I-V) of the dorsal horn in diabetic (STZ-injected) rats compared to control (vehicle-injected) rats. The hybridization signals for NR2A mRNA and NR2B mRNA were significantly elevated in the deep layer of the dorsal horn of diabetic rats. In diabetic (STZ-induced) rats, the levels of expression of mGluR1 mRNA and mGluR5 mRNA were significantly increased in all layers of the dorsal horn. These results suggest that abnormal expression of multiple glutamate receptors is involved in the development of diabetic neuropathy and that glutamate receptors are promising targets in the treatment of this disorder. PMID- 15893612 TI - New live mycobacterial vaccines: the Geneva consensus on essential steps towards clinical development. AB - As the disease caused by Mycobacterium tuberculosis continues to be a burden, which the world continues to suffer, there is a concerted effort to find new vaccines to combat this problem. Of the various vaccines strategies, one viable option is the development of live mycobacterial vaccines. A meeting with researchers, regulatory bodies, vaccines developers and manufactures was held to consider the challenges and progress, which has been achieved with live mycobacterial vaccines (either modified BCG or attenuated M. tuberculosis). Discussion led to the production of a consensus document of the proposed entry criteria for Phase I clinical trials of candidate live mycobacterial vaccines. The vaccine must be characterised thoroughly to prove identity and consistency, as clinical trial lots are prepared. In pre-clinical studies, greater protective efficacy as well as improved safety potential relative to BCG should be considered when assessing potential vaccine candidates. A standard way to measure the protective efficacy to facilitate comparison between vaccine candidates was suggested. Additional safety criteria and verification of attenuation must be considered for attenuated M. tuberculosis. Two non-reverting independent mutations are recommended for such vaccines. When entering Phase I trials, enrollment should be based upon an acceptable characterisation of the study population regarding mycobacterium status and exclude HIV(+) individuals. BCG could be used as a comparator for blinding during the trials and to properly assess vaccine-specific adverse reactions, while assays are being developed to assess immunogenicity of vaccines. The proposed criteria suggested in this consensus document may facilitate the movement of the most promising vaccine candidates to the clinic and towards control of tuberculosis. PMID- 15893613 TI - Development and characterisation of outer membrane vesicle vaccines against serogroup A Neisseria meningitidis. AB - Neisseria meningitidis bacteria of serogroup A are causing recurring meningitis epidemics on the African continent. An outer membrane vesicle (OMV) vaccine against serogroup A meningococci made from a subgroup III serogroup A meningococcal strain was previously shown to induce antibodies with serum bactericidal activity (SBA) in mice. We have here further investigated the properties of OMV vaccines made from five different subgroup III serogroup A meningococcal strains grown in a synthetic medium with low iron content. In addition to the major outer membrane proteins (PorA, PorB, RmpM, Opa and OpcA), small amounts of the NadA, TdfH, Omp85, FetA, FbpA and NspA outer membrane proteins, as well as lipooligosaccharides, were detected in the vaccines. The OMV vaccines were used to immunise mice. Anti-meningococcal IgG antibodies in the mouse sera were analysed by immunoblotting and by enzyme-linked immunosorbent assay against OMVs, and against live meningococcal cells in SBA and a flow cytometric assay. The vaccines induced antibodies with high SBA and opsonophagocytic activity. The strongest IgG responses were directed against PorA. Significant SBA responses were also observed against a subgroup III strain, which did not express PorA, whereas no SBA was observed against a clone IV-1 serogroup A strain. An OMV vaccine from serogroup A meningococci may be an alternative to polysaccharide and conjugate polysaccharide vaccines for Africa. PMID- 15893614 TI - Immunization with pseudorabies virus harboring Fc domain of IgG makes a contribution to protection of mice from lethal challenge. AB - To enhance the efficacy of an inactivated vaccine against pseudorabies virus (PRV), we evaluated the adjuvant properties of Fc domain of IgG. A cell line expressing mouse IgG Fc chimera on its surface was established. We found that when PRV was propagated in the cells expressing the Fc chimera, PRV virion incorporated the Fc. Immunization of BALB/c mice with inactivated PRV harboring Fc, which had been propagated in the cells expressing Fc on its surface, induced higher antibody production against PRV and protected mice more effectively from lethal challenge of virulent strain, comparing to the immunization with normal inactivated virus. Virus harboring Fc has a great potential as a new inactivated vaccine. PMID- 15893615 TI - Identification and characterization of HIV-1-specific CD8+ T cell epitopes presented by HLA-A*2601. AB - Since HLA-A*26 is one of the most common alleles in Asia, where approximately 20% of people have this allele, identification of HIV-1-specific epitopes presented by HLA-A*26 is necessary for studies on the immunopathogenesis of AIDS and vaccine development in Asia. As presented herein, we used the reverse immunogenetics approach to identify HIV-1 epitopes presented by HLA-A*2601, one of the major HLA-A*26 subtypes. We selected 24 HLA-A*2601-binding peptides out of 110 HIV-1 peptides by using a HLA-A*2601 stabilization assay. The ability of these HLA-A*2601-binding peptides to induce peptide-specific CD8(+) T cells was tested by stimulating PBMCs from HIV-1-infected individuals having HLA-A*2601 with these peptides. Four HLA-A*2601-binding peptides induced peptide-specific CD8 T cells. Analysis using HIV-1 recombinant vaccinia-infected C1R-A*2601 cells indicated that these four peptides were HIV-1 epitopes endogenously presented by HLA-A*2601. Two epitope-specific CD8(+) T cells were predominantly detected in HIV-1 infected individuals, suggesting that these epitopes may be useful for vaccine development. PMID- 15893616 TI - Porcine rotavirus strain Gottfried-based human rotavirus candidate vaccines: construction and characterization. AB - Rotavirus gastroenteritis remains the leading cause of severe diarrheal disease in infants and young children worldwide, and thus, a safe and effective rotavirus vaccine is urgently needed in both developing and developed countries. Various candidate rotavirus vaccines that were developed by us and others have been or are being evaluated in different populations in various parts of the world. We have recently confirmed that a porcine rotavirus Gottfried strain bears a P (VP4) serotype (P2B[6]) closely related to human rotavirus P serotype 2A[6] which is of epidemiologic importance in some regions of the world. Based on the modified Jennerian approach to immunization, we have constructed 11 Gottfried-based single VP7 or VP4 gene substitution reassortant vaccine candidates which could provide: (i) an attenuation phenotype of a porcine rotavirus in humans; and (ii) antigenic coverage for G serotypes 1-6 and 8-10 and P serotype 1A[8], 1B[4] and 2A[6]. In addition, following immunization of guinea pigs with Gottfried VP4, we found low but consistent levels of neutralizing antibodies to VP4 with P1A[8] or P1B[4] specificity, both of which are of global epidemiologic importance. Thus, porcine based VP7 reassortant rotavirus vaccines may provide an advantage over rhesus- or bovine-based VP7 reassortant vaccines since the VP4s of the latter vaccines do not evoke antibodies capable of neutralizing the viruses bearing P1A[8], P1B[4] or P2A[6] VP4. PMID- 15893617 TI - Low-frequency ultrasound as a transcutaneous immunization adjuvant. AB - Percutaneous vaccine delivery offers an advantageous mode of immunization due to the unique ability of cutaneous immune cells, especially Langerhans cells, to present antigens to the immune system. Langerhans cells, upon activation, migrate to the regional lymph nodes and lead to the generation of systemic and mucosal immune responses. However, simple topical application of vaccines does not deliver sufficient amounts of antigen in the skin to generate an adequate immune response. Co-administration of strong adjuvants such as cholera toxin or invasive skin abrasion is usually necessary to induce an adequate immune response by topical vaccine application. Here, we report on the use of low-frequency ultrasound as a potent physical adjuvant for successful transcutaneous immunization (TCI). Using tetanus toxoid as a model vaccine, we show that low frequency ultrasound enhances the immune response induced by topical application of tetanus toxoid. The adjuvant effect of ultrasound is partly explained by the enhanced delivery of tetanus toxoid into the skin after ultrasound application and partly by the activation of immune cells after ultrasonic TCI. These studies demonstrate generation of a potent systemic immune response through TCI without using toxin adjuvants or skin abrasion. Ultrasonic TCI offers a needle-free and painless mode of immunization. PMID- 15893618 TI - Recombinant canarypox vectored West Nile virus (WNV) vaccine protects dogs and cats against a mosquito WNV challenge. AB - The safety and efficacy of a canarypox vector expressing PrM and E genes of West Nile virus (WNV) (ALVAC-WNV) was evaluated in dogs and cats. One group of 17 dogs (vaccinated with 10(5.6) TCID(50)) and two groups of cats (groups 1 [n=14] vaccinated with 10(7.5) TCID(50) and 2 [n=8] 10(5.6) TCID(50)) were vaccinated twice at 28-day intervals. Fifteen dogs and eleven cats served as negative controls. The cats and dogs were challenged 120 and 135 days after the second immunization, respectively via the bites of Aedes albopictus mosquitoes infected with WNV. The first dose of vaccine induced a detectable antibody response in four dogs and five cats (one immunized with low and four with high doses). After the second dose, all the vaccinated dogs and all of the cats, immunized with high dose had detectable antibody titers, whereas only four of eight cats in the low dose group were seropositive. None of the vaccinated dogs and one vaccinated cat developed viremia following the WNV mosquito-challenge. In contrast, 14 of the 15 control dogs and 9 of the 11 control cats developed viremia. The experimental vaccine described in this study may be of value in the prevention of WNV infection in dogs and cats. PMID- 15893619 TI - A vector expressing feline mature IL-18 fused to IL-1beta antagonist protein signal sequence is an effective adjuvant to a DNA vaccine for feline leukaemia virus. AB - DNA vaccination using vectors expressing the gag/pol and env genes of feline leukaemia virus (FeLV) and plasmids encoding feline interleukin-12 (IL-12) and IL 18 completely protected cats from viraemia following challenge [Hanlon L, Argyle D, Bain D, Nicolson L, Dunham S, Golder MC, et al. Feline leukaemia virus DNA vaccine efficacy is enhanced by coadministration with interleukin-12 (IL-12) and IL-18 expression vectors. J Virol 2001;75:8424-33]. However, the relative contribution of each cytokine gene towards protection is unknown. This study aimed to resolve this issue. IL-12 and IL-18 constructs were modified to ensure effective expression, and bioactivity was demonstrated using specific assays. Kittens were immunised intramuscularly with FeLV DNA and various cytokine constructs. Together with control kittens, these were challenged oronasally with FeLV and monitored for 15 weeks. All six kittens given FeLV, IL-12 and IL-18 were protected from the establishment of persistent viraemia and four from latent infection. Of six kittens immunised with FeLV DNA and IL-18, all were protected from viraemia and five from latent infection. In contrast, three of five kittens given FeLV DNA and IL-12 became persistently viraemic. Therefore, the adjuvant effect on the FeLV DNA vaccine appears to reside in the expression of IL-18. PMID- 15893620 TI - Immunization of rabbits with Glossina pallidipes tsetse fly midgut proteins: effects on the fly and trypanosome transmission. AB - Proteins isolated from the midgut of Glossina pallidipes were used to immunize rabbits and their efficacy as vaccine candidate(s) against the fly, and their potential to block transmission of Trypanosoma brucei rhodesiense assessed. Two fractions, detergent (DET) and aqueous (AQ) fractions were separated using a non ionic detergent (Triton X-114) and a series of bioassay experiments carried out using serum obtained from rabbits immunized with either of the two fractions. The mortality rates of tsetse flies fed on serum from rabbits immunized with DET and AQ was 56 and 35%, respectively, as compared to 20% mortality in controls. The DET antigen(s) caused considerably higher mortality (chi(2)=1.194, P<0.05) than that on controls. These findings suggest that midgut proteins contain antigens that are lethal to tsetse flies, and are potential candidates for the development of anti-tsetse vaccine. When flies fed on serum derived from DET immunized rabbits were fed on T. b. rhodesiense infected blood, only 20% of them picked the infection. Very few flies (20%) fed on serum derived from DET immunized rabbits had infection of T. b. rhodesiense. In the control flies 45% of them had infection in the midgut with a higher and actively motile parasite load. Assessment of fecundity indicated significantly higher (chi(2)=2.117, P<0.05) larviposition for the control flies when compared to the AQ group of flies (chi(2)=1.054, P<0.05). Significant differences in abortions and pupal weights were also observed. These results suggest that midgut proteins contain antigens with potential for use in development of vaccine to block transmission of trypanosomes through tsetse. PMID- 15893621 TI - Immunogenicity and reactogenicity of combined acellular pertussis/tetanus/low dose diphtheria vaccines given as a booster to UK teenagers. AB - Sustained high incidence of pertussis, particularly amongst unvaccinated infants, is of concern. Inclusion of pertussis vaccination with tetanus and low dose diphtheria (Td) teenage boosters may protect individuals through reproductive years, and prevent transmission to offspring. UK teenagers who had previously received only a three-dose primary course of whole cell pertussis vaccination in infancy and who were due to receive a Td booster (n=323) were randomised to four groups: Td, TdaP, TdaP-inactivated polio vaccine (IPV) (Aventis Pasteur), TdaP (GlaxoSmithKline). There were significant pre- to post-vaccination GMC and GMFR increase for vaccine-contained pertussis antigens (p<0.001) in recipients of aP containing vaccine. All groups demonstrated significant increases pre- to 4 weeks post-vaccination in diphtheria (D)/tetanus (T) geometric mean concentrations (GMCs) and fold rises (GMFRs) (p<0.001). Groups all achieved similar D and T post vaccination GMCs. Local reactogenicity was generally similar between groups and was not associated with pre-booster diphtheria/tetanus antibody levels. A minority of vaccinees reported systemic symptoms with similar proportions between groups for each symptom assessed. This study demonstrated that addition of aP and/or IPV to Td vaccine did not materially alter reactogenicity or immunogenicity of Td components, and induced immune responses to pertussis antigens in teenagers who had received no pertussis vaccine since infancy. PMID- 15893622 TI - Transgene vaccination using Ulex europaeus agglutinin I (UEA-1) for targeted mucosal immunization against HIV-1 envelope. AB - Receptor-mediated gene transfer using an M cell ligand has been shown to be an efficient method for mucosal DNA immunization. To investigate further into alternative M cell ligands, the plant lectin, Ulex europaeus agglutinin I (UEA 1), was tested. UEA-1 binds to human intestinal Caco-2 cells, and these cells can be transfected with poly-l-lysine (PL)-conjugated UEA-1 for expression of reporter cDNAs. When tested in vivo, mice nasally immunized with UEA-1-PL complexed to plasmid encoding HIV-1 envelope showed elevated systemic and mucosal antibody responses, and these were supported by tissue antibody-forming cells. Likewise, elevated envelope-specific CTLs were induced. Thus, UEA-1 mediated DNA delivery represents an alternative mucosal formulation for inducing humoral and cellular immunity against HIV-1. PMID- 15893623 TI - If pneumonia is the "old man's friend", should it be prevented by vaccination? An ethical analysis. AB - Because pneumococcal disease is a major problem among the elderly, pneumococcal polysaccharide vaccination is widely promoted. However, Sir William Osler called pneumonia the friend of the aged, leading to an ethical discussion. Mortality from pneumonia is higher with increasing degrees of underlying illness, outweighing the age effect. Although some symptoms are less common in the elderly, other symptoms are not and the duration may be longer. Problematic criteria for limiting pneumococcal polysaccharide vaccination include age, social value, and quality of life. Recommended criteria for limiting vaccination include autonomous patient refusal, imminent death, and lack of medical benefit, as would be seen in hospice cases. PMID- 15893624 TI - Construction of novel human monoclonal antibodies neutralizing Porphyromonas gingivalis hemagglutination activity using transgenic mice expressing human Ig loci. AB - Porphyromonas gingivalis has been implicated as an important pathogen in the development of adult periodontitis, and its colonization of subgingival sites is critical in the pathogenic process. One potential virulence factor, hemagglutinin, may mediate bacteria attachment onto and penetration into host cells, as well as agglutinate and lyses erythrocytes to intake heme, an absolute requirement for growth. Toward the development of passive immunotherapy, the construction of a human type monoclonal antibody, which is capable of inhibiting the hemagglutinating ability, will be significant and important. The human mAbs, both exhibiting a high degree of specificity and affinity against the recombinant 130 kDa hemagglutinin domain protein have been prepared using XenoMouse technology. The constructed Xeno-mAbs, IgG2 subclass, significantly inhibited hemagglutination of P. gingivalis and its vesicles. The newly constructed Xeno mAbs may prove to be useful for the development of passive immunization against periodontal diseases caused by P. gingivalis infection, pending the results of fertility study in disease mode. PMID- 15893625 TI - Activation of dendritic cells by microparticles containing Bacillus anthracis protective antigen. AB - We have carried out an in vitro investigation into the mechanism by which microencapsulation enhances the immunogenicity of recombinant protective antigen (rPA) from Bacillus anthracis. Murine bone marrow derived dendritic cells (DC) were cocultured with soluble and microencapsulated rPA and the activation status of the cells monitored using FACS. As compared with soluble rPA, it was found that coculture of DC with rPA-loaded microparticles stimulated higher levels of MHC II, CD54, CD80 and CD86 expression (p<0.05). To investigate the longevity of antigen presentation, splenocytes from naive mice were pulsed overnight with (3)H thymidine following 1, 3 or 6 days coculture with DC transiently exposed to soluble or microencapsulated rPA. Splenocyte proliferation was more pronounced, and continued for a more protracted period, if the 'feeder' cells were exposed to microencapsulated antigen as compared with soluble antigen or 'empty' microspheres. To this end, our findings indicate that microsphere uptake increases the surface expression of MHC and co-stimulatory molecules on DC and can facilitate prolonged presentation of antigen to T-cells, possibly by acting as an intracellular depot. PMID- 15893626 TI - Characterization of DNA vaccines encoding the domains of calreticulin for their ability to elicit tumor-specific immunity and antiangiogenesis. AB - Antigen-specific cancer immunotherapy and antiangiogenesis are feasible strategies for cancer therapy because they can potentially treat systemic tumors at multiple sites in the body while discriminating between neoplastic and non neoplastic cells. We have previously developed a DNA vaccine encoding calreticulin (CRT) linked to human papillomavirus-16 E7 and have found that this vaccine generates strong E7-specific antitumor immunity and antiangiogenic effects in vaccinated mice. In this study, we characterized the domains of CRT to produce E7-specific antitumor immunity and antiangiogenic effects by generating DNA vaccines encoding each of the three domains of CRT (N, P, and C domains) linked to the HPV-16 E7 antigen. We found that C57BL/6 mice vaccinated intradermally with DNA encoding the N domain of CRT (NCRT), the P domain of CRT (PCRT), or the C domain of CRT (CCRT) linked with E7 exhibited significant increases in E7-specific CD8(+) T cell precursors and impressive antitumor effects against E7-expressing tumors compared to mice vaccinated with wild-type E7 DNA. In addition, the N domain of CRT also showed antiangiogenic properties that might have contributed to the antitumor effect of NCRT/E7. Thus, the N domain of CRT can be linked to a tumor antigen in a DNA vaccine to generate both antigen-specific immunity and antiangiogenic effects for cancer therapy. PMID- 15893627 TI - Regional hippocampal alteration associated with cognitive deficit following experimental brain injury: a systems, network and cellular evaluation. AB - Cognitive deficits persist in patients who survive traumatic brain injury (TBI). Lateral fluid percussion brain injury in the mouse, a model of human TBI, results in hippocampal-dependent cognitive impairment, similar to retrograde amnesia often associated with TBI. To identify potential substrates of the cognitive impairment, we evaluated regional neuronal loss, regional hippocampal excitability and inhibitory synaptic transmission. Design-based stereology demonstrated an approximate 40% loss of neurons through all subregions of the hippocampus following injury compared with sham. Input/output curves recorded in slices of injured brain demonstrated increased net synaptic efficacy in the dentate gyrus in concert with decreased net synaptic efficacy and excitatory postsynaptic potential-spike relationship in area CA1 compared with sham slices. Pharmacological agents modulating inhibitory transmission partially restored regional injury-induced alterations in net synaptic efficacy. Both evoked and spontaneous miniature inhibitory postsynaptic currents (mIPSCs) recorded in surviving dentate granule neurons were smaller and less frequent in injured brains than in uninjured brains. Conversely, both evoked and spontaneous mIPSCs recorded in surviving area CA1 pyramidal neurons were larger in injured brains than in uninjured brains. Together, these alterations suggest that regional hippocampal function is altered in the injured brain. This study demonstrates for the first time that brain injury selectively disrupts hippocampal function by causing uniform neuronal loss, inhibitory synaptic dysfunction, and regional, but opposing, shifts in circuit excitability. These changes may contribute to the cognitive impairments that result from brain injury. PMID- 15893628 TI - Cytokine modulation of defensive rage behavior in the cat: role of GABAA and interleukin-2 receptors in the medial hypothalamus. AB - Defensive rage behavior is a form of aggressive behavior occurring in nature in response to a threatening stimulus. It is also elicited by stimulation of the medial hypothalamus and midbrain periaqueductal gray (PAG) and mediated through specific neurotransmitter-receptor mechanisms within these regions. Since interleukin (IL)-2 modulates the release of neurotransmitters linked to aggression and rage, we sought to determine whether IL-2 microinjected into the medial hypothalamus would modulate defensive rage. Microinjections of relatively low doses of IL-2 into the medial hypothalamus significantly suppressed defensive rage elicited from the PAG in a dose-dependent manner and in the absence of signs of sickness behavior. Pre-treatment with an antibody directed against IL-2Ralpha or a GABA(A) receptor antagonist blocked IL-2's suppressive effects upon defensive rage. Since the suppression of defensive rage is also mediated by 5 HT(1) receptors in the medial hypothalamus, a 5-HT(1) antagonist was microinjected into this region as a pretreatment for IL-2; however, it did not block IL-2's suppressive effects. Immunocytochemical data provided anatomical support for these findings by revealing extensive labeling of IL-2Ralpha on neurons in the medial hypothalamus. IL-2 microinjected into the medial hypothalamus did not modulate predatory attack elicited from the lateral hypothalamus. In summary, we provide evidence for a novel role for IL-2 in the medial hypothalamus as a potent suppressor of defensive rage behavior. These effects are mediated through an IL-2-GABA(A) receptor mechanism. PMID- 15893629 TI - Computation of inverse functions in a model of cerebellar and reflex pathways allows to control a mobile mechanical segment. AB - The command and control of limb movements by the cerebellar and reflex pathways are modeled by means of a circuit whose structure is deduced from functional constraints. One constraint is that fast limb movements must be accurate although they cannot be continuously controlled in closed loop by use of sensory signals. Thus, the pathways which process the motor orders must contain approximate inverse functions of the bio-mechanical functions of the limb and of the muscles. This can be achieved by means of parallel feedback loops, whose pattern turns out to be comparable to the anatomy of the cerebellar pathways. They contain neural networks able to anticipate the motor consequences of the motor orders, modeled by artificial neural networks whose connectivity is similar to that of the cerebellar cortex. These networks learn the direct biomechanical functions of the limbs and muscles by means of a supervised learning process. Teaching signals calculated from motor errors are sent to the learning sites, as, in the cerebellum, complex spikes issued from the inferior olive are conveyed to the Purkinje cells by climbing fibers. Learning rules are deduced by a differential calculation, as classical gradient rules, and they account for the long term depression which takes place in the dendritic arborizations of the Purkinje cells. Another constraint is that reflexes must not impede voluntary movements while remaining at any instant ready to oppose perturbations. Therefore, efferent copies of the motor orders are sent to the interneurones of the reflexes, where they cancel the sensory-motor consequences of the voluntary movements. After learning, the model is able to drive accurately, both in velocity and position, angular movements of a rod actuated by two pneumatic McKibben muscles. Reflexes comparable to the myotatic and tendinous reflexes, and stabilizing reactions comparable to the cerebellar sensory-motor reactions, reduce efficiently the effects of perturbing torques. These results allow to link the behavioral concepts of the equilibrium-point "lambda model" [J Motor Behav 18 (1986) 17] with anatomical and physiological features: gains of reflexes and sensori-motor reactions set the slope of the "invariant characteristic," and efferent copies set the "threshold of the stretch reflex." Thus, mathematical and physical laws account for the raison d'etre of the inhibitory nature of Purkinje cells and for the conspicuous anatomical pattern of the cerebellar pathways. These properties of these pathways allow to perform approximate inverse calculations after learning of direct functions, and insure also the coordination of voluntary and reflex motor orders. PMID- 15893630 TI - Neurotrophin-3 is a target-derived neurotrophic factor for penile erection inducing neurons. AB - The aim of this study was to determine whether the neurotrophins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin (NT)-3 could act as endogenous target-derived trophic factors for erection-inducing, i.e. penis-projecting major pelvic ganglion (MPG) neurons, and/or penile sensory neurons in adult rat. This was accomplished by studying the expression of NT mRNAs in the penis and their cognate receptors in the MPG and dorsal root ganglia (DRGs), and the retrograde axonal transport of radioiodinated NTs injected into the corpora cavernosa. Northern hybridization showed that NGF, BDNF, and NT-3 mRNAs are expressed in the shaft of the penis. In situ hybridization combined with usage of the retrograde tracer Fluoro-Gold showed that TrkC and p75 receptors are expressed in penis-projecting neurons of the MPG whereas the mRNAs for TrkA and TrkB receptors were undetectable. However, all the NT receptor mRNAs were expressed in penile sensory neurons of sacral level 1 (S1) DRG. (125)I-NT-3 injected into the shaft of the penis was retrogradely transported into the MPG and S1 DRG, whereas radioiodinated NGF and BDNF were transported specifically into the S1 DRG, thus confirming the existence of functional NT receptors in these penile neurons. In conclusion, these data suggest that NT-3 may act as a target-derived neurotrophic factor for both erection-inducing and penile sensory neurons, whereas NGF and BDNF may be more important for the sensory innervation of the penis. PMID- 15893631 TI - Generation and phenotypic characterization of a galanin overexpressing mouse. AB - In most parts of the peripheral nervous system galanin is expressed at very low levels. To further understand the functional role of galanin, a mouse overexpressing galanin under the platelet-derived growth factor-B was generated, and high levels of galanin expression were observed in several peripheral tissues and spinal cord. Thus, a large proportion of neurons in autonomic and sensory ganglia were galanin-positive, as were most spinal motor neurons. Strong galanin like immunoreactivity was also seen in nerve terminals in the corresponding target tissues, including skin, blood vessels, sweat and salivary glands, motor end-plates and the gray matter of the spinal cord. In transgenic superior cervical ganglia around half of all neuron profiles expressed galanin mRNA but axotomy did not cause a further increase, even if mRNA levels were increased in individual neurons. In transgenic dorsal root ganglia galanin mRNA was detected in around two thirds of all neuron profiles, including large ones, and after axotomy the percentage of galanin neuron profiles was similar in overexpressing and wild type mice. Axotomy reduced the total number of DRG neurons less in overexpressing than in wild type mice, indicating a modest rescue effect. Aging by itself increased galanin expression in the superior cervical ganglion in wild type and transgenic mice, and in the latter also in preganglionic cholinergic neurons projecting to the superior cervical ganglion. Galanin overexpressing mice showed an attenuated plasma extravasation, an increased pain response in the formalin test, and changes in muscle physiology, but did not differ from wild type mice in sudomotor function. These findings suggest that overexpressed galanin in some tissues of these mice can be released and via a receptor-mediated action influence pathophysiological processes. PMID- 15893632 TI - Adenosine A1 and A2A receptors are co-expressed in pyramidal neurons and co localized in glutamatergic nerve terminals of the rat hippocampus. AB - Adenosine is a neuromodulator that controls neurotransmitter release through inhibitory A1 and facilitatory A2A receptors. Although both adenosine receptor mediated inhibition and facilitation of glutamate release have been observed, it is not clear whether both A1 and A2A receptors are located in the same glutamatergic nerve terminal or whether they are located on different populations of these terminals. Thus, we have tested if single pyramidal glutamatergic neurons from the hippocampus simultaneously expressed A1 and A2A receptor mRNA and if A1 and A2A receptors were co-localized in hippocampal glutamatergic nerve terminals. Single cell PCR analysis of visually identified pyramidal neurons revealed the simultaneous presence of A1 and A2A receptor mRNA in four out 16 pyramidal cells possessing glutamatergic markers but not GABAergic or astrocytic markers. Also, A1 and A2A receptor immunoreactivities were co-localized in 26 +/- 4% of nerve terminals labeled with antibodies against vesicular glutamate transporters type 1 or 2, i.e. glutamatergic nerve terminals. This indicates that glutamatergic neurons in the hippocampus co-express A1 and A2A receptors and that these two receptors are co-localized in a subset of glutamatergic nerve terminals. PMID- 15893633 TI - Functional neural stem cells derived from adult bone marrow. AB - Pluripotent hematopoietic cells from adult bone marrow may give rise not only to neurons, oligodendrocytes and astrocytes after transplantation into newborn brains, but also to neural stem cells (NSC). These NSC localize to both the ventricular epithelium and subventricular zone, persist in the transplanted brain, and may generate neurospheres 1 month after transplant, which after in vitro expansion differentiate into the different neural lineages. Furthermore, the bone marrow-derived NSC differentiate in vivo into functional oligodendrocytes and neurons following demyelinating lesions, thus, demonstrating the ability of adult bone marrow progenitors to generate self-renewing, functional neural stem cells, validating this approach as an alternative source of long-lasting neural stem cells with therapeutic implications in neurodegenerative diseases. PMID- 15893634 TI - Mu-opioid-mediated inhibition of glutamate synaptic transmission in rat central amygdala neurons. AB - The central nucleus of the amygdala (CeA) plays an important role both in stimulus-reward learning for the reinforcing effects of drugs of abuse and in environmental condition-induced analgesia. Both of these two CeA functions involve the opioid system within the CeA. However, the pharmacological profiles of its opioid receptor system have not been fully studied and the synaptic actions of opioid receptors in the CeA are largely unknown. In this study with whole-cell voltage-clamp recordings in brain slices in vitro, we examined actions of opioid agonists on glutamate-mediated excitatory postsynaptic currents (EPSCs) in CeA neurons. Opioid peptide methionine-enkephalin (ME; 10 microM) produced a significant inhibition (38%) in the amplitude of evoked EPSCs, an action mimicked by the mu-opioid receptor agonist [D-Ala(2),N-MePhe(4),Gly-ol(5)]-enkephalin (DAMGO; 1 microM, 44%). Both effects of ME and DAMGO were abolished by the mu receptor antagonist CTAP (1 microM), suggesting a mu receptor-mediated effect. Neither delta-opioid receptor agonist [D-Pen(2),D-Pen(5)]-enkephalin (1 microM) nor kappa-opioid receptor agonist U69593 (300 nM) had any effect on the glutamate EPSC. ME significantly increased the paired-pulse ratio of the evoked EPSCs and decreased the frequency of miniature EPSCs without altering the amplitude of miniature EPSCs. Furthermore, the mu-opioid inhibition of the EPSC was blocked by 4-aminopyridine (4AP; 100 microM), a voltage-dependent potassium channel blocker, and by phospholipase A(2) inhibitors AACOCF(3) (10 microM) and quinacrine (10 microM). These results indicate that only the mu-opioid receptor is functionally present on presynaptic glutamatergic terminals in normal CeA neurons, and its activation reduces the probability of glutamate release through a signaling pathway involving phospholipase A(2) and the presynaptic, 4AP-sensitive potassium channel. This study provides evidence for the presynaptic regulation of glutamate synaptic transmission by mu-opioid receptors in CeA neurons. PMID- 15893635 TI - Comparative expression profiles of ShcB and ShcC phosphotyrosine adapter molecules in the adult brain. AB - Shc family of adaptor molecules has been demonstrated to play an important role during the transition from proliferating neural stem cells to postmitotic neurons. Previous studies from our group demonstrated a progressive decrease of ShcA levels occurring in coincidence with the end of embryonic neurogenesis and neuronal maturation, being ShcB and ShcC the major Shc molecules expressed in the mature brain. A growing body of evidence indicates that ShcB and ShcC are neuronal specific molecules exerting important roles in neuronal survival and phenotypic stability thus becoming potential attracting target molecules for development of drugs for interfering with brain demises. Here, we examine the expression pattern of ShcB and ShcC in neuronal populations composing the adult central and peripheral nervous system, in order to better elucidate their roles in vivo. We found a heterogeneous and peculiar presence and subcellular localization of ShcB and ShcC in specific neuronal populations, enlightening a potential specific requirement of these two molecules in the survival/maintenance of defined neuronal subtypes. PMID- 15893636 TI - Rotenone induces aggregation of gamma-tubulin protein and subsequent disorganization of the centrosome: relevance to formation of inclusion bodies and neurodegeneration. AB - Neurodegenerative disorders are characterized by progressive loss of specific neurons in the central nervous system. Although they have different etiologies and clinical manifestations, most of them share similar histopathologic characteristics such as the presence of inclusion bodies in both neurons and glial cells, which represent intracellular aggregation of misfolded or aberrant proteins. In Parkinson's disease, formation of inclusion bodies has been associated with the aggresome-related process and consequently with the centrosome. However, the significance of the centrosome in the neurodegenerative process remains obscure. In the present study, the morphological and functional changes in the centrosome induced by rotenone, a common insecticide used to produce experimental Parkinsonism, were examined both in vitro and in vivo. Aggregation of gamma-tubulin protein, which is a component of the centrosome matrix and recently identified in Lewy bodies of Parkinson's disease, was observed in primary cultures of mesencephalic cells treated with rotenone. Rotenone-treated neurons and astrocytes showed enlarged and multiple centrosomes. These centrosomes also displayed multiple aggregates of alpha-synuclein protein. Neurons with disorganized centrosomes exhibited neurite retraction and microtubule destabilization, and astrocytes showed disturbances of mitotic spindles. The Golgi apparatus, which is closely related to the centrosome, was dispersed in both rotenone-treated neuronal cells and the substantia nigra of rotenone-treated rats. Our findings suggested that recruitment of abnormal proteins in the centrosome contributed to the formation of inclusion bodies, and that rotenone markedly affected the structure and function of the centrosome with consequent induction of cytoskeleton disturbances, disassembly of the Golgi apparatus and collapse of neuronal cells. PMID- 15893637 TI - Movements restriction and alterations of the number of spines distributed along the apical shafts of layer V pyramids in motor and primary sensory cortices of the peripubertal and adult rat. AB - The number and distribution of spines along apical shafts of rapid-Golgi-stained layer V pyramidal cells from visual, motor and somatosensory cortical areas were analyzed in control and movement-restricted (beginning at 20 days old) Wistar rats killed at 30, 40, 80 and 120 days of age (experiment A). In other group of rats, spine density was analyzed when restriction initiates on day 40 and the animals were killed at 50, 60 and 80 days postpartum, or after restriction starting on day 80 and killed at 120 days of age (experiment B). It has been found that the restriction of movements significantly reduces the total number of spines on apical shafts in the three cortical areas, when this condition starts at 20, 40 or 80 days without changing the overall distribution of spines. Also present findings indicate that the effects of movements restriction are attenuated when they were concurrent with maturational brain processes (20-40 days) than when they occurred later in life. The question remains open of which part of the measured reduction on the number of spines is due to the immobilization and which to the stress associated with this maneuver. PMID- 15893638 TI - Pregnenolone and dehydroepiandrosterone administration in neonatal rats alters the immunoreactivity of hippocampal synapsin I, neuropeptide Y and glial fibrillary acidic protein at post-puberty. AB - It is well documented that neurosteroids administered during the neonatal period influence the development of several brain systems. In our previous study, pregnenolone administered to rats during the neonatal period altered adenosinergic and dopaminergic functions in the striatum and cerebral cortex. The present study examined the effects of the treatment with pregnenolone and dehydroepiandrosterone (DHEA) from the postnatal day (P) 3-P7 on synapsin I (a marker for presynaptic terminals) and glial fibrillary acidic protein (GFAP: a marker for astroglia) levels in the hippocampus of Sprague-Dawley rats at 3 and 7 weeks of age. In addition, neuropeptide Y and dynorphin A immunoreactivity was measured. The administration of pregnenolone and DHEA to neonatal rats significantly altered the expression of synapsin I in the dentate gyrus and CA3 region at post-puberty but not at pre-puberty. A significantly greater expression of GFAP-immunoreactive astrocytes or processes was demonstrated in the pregnenolone- and DHEA-treated groups at both pre-puberty and post-puberty. A significant increase in the number and size of GFAP-immunoreactive astrocytes and in the extension of arborization was seen in the overall hippocampus. The number of neuropeptide Y-positive cells in the hilus region was also significantly increased in the neurosteroid-treated group at post-puberty. No differences were detected in dynorphin A immunoreactivity among the experimental groups. These results of this study suggest that pregnenolone and DHEA play an important role in the development of hippocampus. PMID- 15893639 TI - Minocycline attenuates lipopolysaccharide-induced white matter injury in the neonatal rat brain. AB - Our previous studies have shown that intracerebral administration of endotoxin, lipopolysaccharide (LPS), induces selective white matter injury and hypomyelination in the neonatal rat brain and that the LPS-induced brain injury is associated with activation of microglia. To test the hypothesis that inhibition of microglial activation may protect against LPS-induced white matter injury, we examined roles of minocycline, a putative suppressor of microglial activation, on LPS-induced brain injury in the neonatal rat. A stereotactic intracerebral injection of LPS (1 mg/kg) was performed in postnatal day 5 Sprague Dawley rats and control rats were injected with sterile saline. Minocycline (45 mg/kg) was administered intraperitoneally 12 h before and immediately after LPS injection and then every 24 h for 3 days. Inflammatory responses, activation of microglia and brain injury were examined 1 and 3 days after LPS injection. LPS injection resulted in brain injury in selective brain areas, including bilateral ventricular enlargement, cell death at the sub- and periventricular areas, loss of O4+ and O1+ oligodendrocyte (OL) immunoreactivity and hypomyelination, as indicated by decreased myelin basic protein immunostaining, in the neonatal rat brain. Minocycline administration significantly attenuated LPS-induced brain injury in these rat brains. The protective effect of minocycline was associated with suppressed microglial activation as indicated by the decreased number of activated microglial cells following LPS stimulation and with consequently decreased elevation of interleukin 1beta and tumor necrosis factor-alpha concentrations induced by LPS and a reduced number of inducible nitric oxide synthase expressing cells. Protection of minocycline was also linked with the reduction in LPS-induced oxidative stress, as indicated by 4-hydroxynonenal positive OLs. The overall results suggest that reduction in microglial activation may protect the neonatal brain from LPS-induced white matter injury and inhibition of microglial activation might be an effective approach for the therapeutic treatment of infection-induced white matter injury. PMID- 15893640 TI - Nerve-induced release of nitric oxide from the rabbit corpus cavernosum is modulated by cyclic guanosine 3',5'-monophosphate. AB - Nitric oxide (NO) is a neurotransmitter of the autonomic nerves in the urogenital tract, in particular the release of NO in the cavernous tissue is of importance for maintaining erection. However, the regulation of NO formation in neurons of the corpus cavernosum is poorly understood. Here, we report, that upon electrical stimulation of isolated rabbit corpus cavernosum, NO/NO(2-) was formed and released in a reproducible fashion. The NO synthase inhibitor N(omega)-nitro-L arginine methyl ester decreased the amount of NO/NO(2-) released to 50+/-18% (P<0.05). The neurotoxin tetrodotoxin diminished the nerve-induced release of NO/NO(2-), to 35+/-10% (P<0.001). Blockage of the cholinergic and noradrenergic pathways by application of scopolamine and guanethidin (both 10(-5) M) did not alter the basal or nerve-evoked formation of NO/NO(2-). We also applied modulators of the soluble guanylate cyclase (sGC)/cyclic guanosine 3',5' monophosphate (cGMP) pathway to study if and to what extent cGMP might affect the release of NO from the erectile tissue. In the presence of the cGMP analog 8-Br cGMP (10(-4) M), and, the sGC stimulator 3-(5'-hydroxymethyl-2'-furyl)-1 benzylindazole (10(-4) M), the release of NO/NO(2-) was increased to 385+/-120% (P<0.05) and 282+/-78% (P<0.05), respectively. The effect of the phosphodiesterase inhibitor zaprinast (10(-4) M), was not significant (209+/-53%, n.s). In contrast, inhibition of sGC by 1-H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1 one (10(-5) M) decreased the release of NO/NO(2-) to 64+/-14% (P<0.05). Our results suggest that NO/NO(2-) is released by nitrergic neurons within the rabbit corpus cavernosum and that the release is subject to modulation by the sGC/cGMP pathway, but not to modulation by acetylcholine or noradrenaline. PMID- 15893641 TI - Extrasynaptic localization of glycine receptors in the rat supraoptic nucleus: further evidence for their involvement in glia-to-neuron communication. AB - Neurons of the rat supraoptic nucleus (SON) express glycine receptors (GlyRs), which are implicated in the osmoregulation of neuronal activity. The endogenous agonist of the receptors has been postulated to be taurine, shown to be released from astrocytes. We here provide additional pieces of evidence supporting the absence of functional glycinergic synapses in the SON. First, we show that blockade of GlyRs with strychnine has no effect on either the amplitude or frequency of miniature inhibitory postsynaptic currents recorded in SON neurons, whereas they were all suppressed by the GABA(A) antagonist gabazine. Then, double immunostaining of sections with presynaptic markers and either GlyR or GABA(A) receptor (GABA(A)R) antibodies indicates that, in contrast with GABA(A)Rs, most GlyR membrane clusters are not localized facing presynaptic terminals, indicative of their extrasynaptic localization. Moreover, we found a striking anatomical association between SON GlyR clusters and glial fibrillary acidic protein (GFAP) positive astroglial processes, which contain high levels of taurine. This type of correlation is specific to GlyRs, since GABA(A)R clusters show no association with GFAP-positive structures. These results substantiate and strengthen the concept of extrasynaptic GlyRs mediating a paracrine communication between astrocytes and neurons in the SON. PMID- 15893642 TI - Chronic exposure to ammonia alters the modulation of phosphorylation of microtubule-associated protein 2 by metabotropic glutamate receptors 1 and 5 in cerebellar neurons in culture. AB - Hyperammonemia impairs signal transduction associated to glutamate receptors and phosphorylation of some neuronal proteins including microtubule-associated protein 2 (MAP-2). The aim of this work was to analyze the effects of hyperammonemia on modulation of MAP-2 phosphorylation by metabotropic glutamate receptors (mGluRs) in rat cerebellar neurons in culture. Hyperammonemia increased basal phosphorylation of MAP-2 (180%). Activation of mGluRs 1 and 5 with (S)-3,5 dihydroxyphenylglycine (DHPG) increased MAP-2 phosphorylation (170%) in control neurons but not in neurons exposed to ammonia. Activation of mGluRs 2 and 3 with (2S,3S,4S)-CCG/(2S, 1'S,2'S)-2-(carboxycyclopropyl)glycine increased slightly (25%) MAP-2 phosphorylation in neurons exposed to ammonia or not. Activation of mGluR5 with (+/-)-trans-azetidine-2,4-dicarboxylic acid increased MAP-2 phosphorylation (24%) in control neurons but decreased it by 56% in neurons exposed to ammonia. Activation of mGluR1 using 2-methyl-6-(phenylethynyl)pyridine and DHPG increased MAP-2 phosphorylation 183% in control neurons but only 89% in neurons exposed to ammonia. In control neurons mGluR1 activation greatly increases phosphorylation of MAP-2, while activation of mGluRs 5, 2 or 3 increased it slightly. Taken together, hyperammonemia reduces the increase in MAP 2 phosphorylation induced by mGluR1activation. Moreover, in neurons exposed to ammonia activation of mGluR5 reduces MAP-2 phosphorylation. These effects reflect significant alterations in signal transduction associated to mGluR1 and mGluR5 in hyperammonemia that may contribute to altered glutamatergic neurotransmission and to the neurological alterations in hyperammonemia and hepatic encephalopathy. PMID- 15893643 TI - Intrinsic connections of the cingulate cortex in the rat suggest the existence of multiple functionally segregated networks. AB - The cingulate cortex is a functionally and morphologically heterogeneous cortical area comprising a number of interconnected subregions. To date, the exact anatomy of intracingulate connections has not been studied in detail. In the present study we aimed to determine the topographical and laminar characteristics of intrinsic cingulate connections in the rat, using the anterograde tracers Phaseolus vulgaris-leucoagglutinin and biotinylated dextran amine. For assessment of these data we further refined and compared the existing cytoarchitectonic descriptions of the two major cingulate constituents, the anterior cingulate and retrosplenial cortices. The results of this study demonstrate that rostral areas, i.e. the infralimbic and prelimbic cortices and the rostral one third of the dorsal anterior cingulate cortex are primarily interconnected with each other and not with other cingulate areas. The caudal two thirds of the dorsal anterior cingulate cortex project to the caudal part of the ventral anterior cingulate cortex, whereas the entire ventral anterior cingulate cortex projects to only the mid-rostro-caudal part of the dorsal anterior cingulate cortex. Dense reciprocal connections exist between the remaining, i.e. the supracallosal parts of the anterior cingulate and retrosplenial cortices with a general rostro-caudal topography, in the sense that the rostral part of the anterior cingulate cortex and caudal part of the retrosplenial cortex are interconnected and the same holds true for the caudal part of the anterior cingulate cortex and rostral part of the retrosplenial cortex. This topographical pattern of intracingulate connections relates to the results of several functional studies, suggesting that specific cingulate functions depend on a number of interconnected cingulate subregions. Through their intricate associational connections, these subregions form functionally segregated networks. PMID- 15893644 TI - Identification and characterization of six new alternatively spliced variants of the human mu opioid receptor gene, Oprm. AB - The mu opioid receptor plays an important role in mediating the actions of morphine and morphine-like drugs. Receptor binding and a wide range of pharmacological studies have proposed several mu receptor subtypes, but only one mu opioid receptor (Oprm) gene has been isolated. Like the mouse and rat, the human Oprm gene undergoes alternative splicing. In the present studies, we have identified and characterized six new splice variants from the human Oprm gene using a reverse transcription-polymerase chain reaction strategy, yielding a total of 10 human splice variants of the mu opioid receptor MOR-1. All the variants identified contained exons 1, 2 and 3, but differed from MOR-1 itself and each other by splicing downstream from exon 3, resulting in different amino acid sequences. Northern blot analysis demonstrated expression of the variant mRNAs. Receptor binding assays established that these variants belonged to the mu opioid receptor family with limited differences in mu opioid ligand affinities and selectivity. However, adenylyl cyclase and [35S]GTPgammaS binding assays revealed major differences in both potency and efficacy among these variants. The dissociation between binding affinity, potency and efficacy for the opioids among these variants may provide insights into the wide range of opioid responses among these agents observed clinically and opens new avenues in designing selective drugs based upon their efficacy and potency rather simple binding affinity. PMID- 15893645 TI - Antenatal glucocorticoids blunt the functioning of the hypothalamic-pituitary adrenal axis of neonates and disturb some behaviors in juveniles. AB - Antenatal glucocorticoids are highly effective in preventing respiratory distress of premature babies but can induce physiological and behavioral disturbances in young infants as well as in animals. Therefore, the hypothalamic-pituitary adrenal (HPA) axis of rat neonates, and the consequences on behavioral development of offspring have been studied after five antenatal injections of dexamethasone (DEX) or vehicle. DEX decreased offspring body weight at birth, and significantly delayed the normal growth for the first 3 weeks of life. This paralleled diminished behavioral performances measured on postnatal day 3 (righting reflex) and postnatal day 10 (grasping test). Circulating levels of adrenocorticotrophin (ACTH) and corticosterone were significantly decreased on postnatal day 1 and this was related to a diminution of HPA axis activity shown by the decrease of central expression of corticotropin releasing hormone (CRH) mRNA, immunoreactive content in paraventricular neurons (PVN) and in the median eminence endings were significantly decreased. On the other hand, expression of another secretagogue of ACTH, arginine vasopressin (AVP), was differently affected in the PVN parvocellular neurons of offspring of the DEX group since AVP mRNA increased whereas immunoreactive content of the PVN parvocellular neurons was lowered. Simultaneously, the co-production of AVP and CRH in PVN neurons was stimulated. This can support the view that antenatal DEX reached the fetus and produced some damage which did not parallel that induced by prenatal stress of the pregnant females, especially the low body weight of offspring. The harmful consequence of antenatal DEX treatment was not restrictively due to the blunting of the HPA axis but also to the low body weight, which disturbed behavioral performances for the first weeks of life and could participate in other disorders in adult life. PMID- 15893646 TI - A brainstem substrate for analgesia elicited by intraoral sucrose. AB - Previous studies demonstrated that nursing or intraoral infusion of certain components of mother's milk (e.g. sugars and fats) produces calming and opiate receptor-dependent analgesia in newborn rats and humans. However, the neural circuitry underlying such analgesia is unknown. The aim of the present study was to specify the central pathways by which taste stimuli engage neural antinociceptive mechanisms. For this purpose, midcollicular transactions were used to investigate the role of the forebrain in analgesia elicited by intraoral infusion of 0.2 M sucrose in neonatal rats. Sucrose-induced analgesia persisted, and was enhanced, following midcollicular transection, indicating that it did not require neural circuits confined to the forebrain. Fos immunohistochemistry was used to identify brainstem neurons activated by a brief (90 s) intraoral infusion of a small volume (90 microl, 0.2M) of sucrose or a salt solution (0.1 M ammonium chloride) in 10-day-old rat pups. Compared with control groups (intact, cannula, distilled water), both sucrose and ammonium chloride induced Fos expression in the rostral nucleus tractus solitarius, the first relay in the ascending gustatory pathway. Sucrose also elicited Fos expression in several brainstem areas associated with centrally mediated analgesia, including the periaqueductal gray and the nucleus raphe magnus. Taken together, these findings demonstrate that analgesia elicited by intraoral sucrose does not require involvement of the forebrain. Intraoral sucrose activates neurons in the periaqueductal gray and nucleus raphe magnus, two key brainstem sites critically involved in descending pain modulation. PMID- 15893647 TI - Painful stimuli induce in vivo phosphorylation and membrane mobilization of mouse spinal cord NKCC1 co-transporter. AB - The Na+ --Cl- --K+ isoform 1 (NKCC1) is a co-transporter that increases the intracellular concentration of chloride. NKCC1 plays a critical role in neuronal excitability and it has been recently suggested that it can contribute to hyperalgesic states by modulating the chloride concentration inside nociceptive neurons. In the spinal cord, trafficking of neurotransmitter receptors from the cytosol to the plasma membrane has been demonstrated to contribute to the development of hyperalgesia. However, it is unknown if trafficking of co transporters can also occur in the nervous system or if it can be induced by painful stimulation. In this study, we have induced referred mechanical hyperalgesia in vivo by intracolonic instillation of capsaicin in mice. Using subcellular fractionation of proteins and cross-linking of membrane proteins we have observed that intracolonic capsaicin induced a 50% increase in NKCC1 in the plasma membrane of lumbosacral spinal cord 90 and 180 min after instillation, in parallel with a similar decrease in the cytosolic fraction. These effects returned to basal levels 6 h after capsaicin treatment. Intracolonic capsaicin also evoked a rapid (10 min) and transient phosphorylation of NKCC1, however, intracolonic saline did not produce significant changes in either NKCC1 trafficking or phosphorylation and none of the treatments induced any alterations of NKCC1 in the thoracic spinal cord. These results suggest that phosphorylation and recruitment of NKCC1 might play a role in referred mechanical hyperalgesia evoked by a painful visceral stimulus. The time course of the effects observed suggests that phosphorylation could contribute to the initial generation of hyperalgesia whereas trafficking could participate in the maintenance of hyperalgesic states observed at longer time points. PMID- 15893648 TI - Three-dimensional culture of newborn rat utricle using an extracellular matrix promotes formation of a cyst. AB - The vestibule is the end organ devoted to sensing of head movements in space. To function properly, its mechano-receptors require the presence of a unique apical extracellular medium, the endolymph. Numerous studies have elucidated the mechanisms involved in the production and homeostasis of this unique medium and the responses of sensory cells to stimulation. However, anatomical constraints have prevented direct and simultaneous studies of their relationships. The aim of this study was the development of an in vitro model that would allow concomitant investigations on maturation and physiological properties of both the hair cells and their endolymphatic compartment. A three-dimensional (3D) culture of newborn rat utricles using an extracellular matrix sustaining 3D cellular growth was developed during 3, 6, or 10 days in vitro (DIV). Using morphological and electrophysiological techniques, we describe the de novo formation of a cyst. It was composed of the sensory epithelium and non-sensory cells-canalar, dark and intermediate cells-that polarized so that their apical surface faced its lumen. During the time of culture, the utricular potential (UP) was steady (-1.1+/-5.0 mV) in oxygenated condition, while in anoxia, the UP significantly decreased to 8.4+/-1.0 mV at 8 DIV. Over the same period, the K+ concentration in the cyst increased up to 86.1+/-33.9 mM (versus 5.6+/-1.5 mM in the bath). These observations indicated that the mechanisms generating the UP and the K-secretory activity were functional at this stage. Concomitantly, the hair cells acquired mature and functional properties: the type 1 and type 2 phenotypes, a mean resting membrane potential of -68.1+/-4.6 mV and typical electrophysiological responses. This preparation provides a powerful means to simultaneous access the hair cells and their endolymphatic compartment, with the possibility to use multi technical approaches to investigate their interdependent relationships. PMID- 15893649 TI - Role of neuropeptide Y in the regulation of gonadotropin releasing hormone system in the forebrain of Clarias batrachus (Linn.): immunocytochemistry and high performance liquid chromatography-electrospray ionization-mass spectrometric analysis. AB - Although the importance of neuropeptide Y (NPY) in the regulation of gonadotropin releasing hormone (GnRH) and reproduction has been highlighted in recent years, the neuroanatomical substrate within which these substances might interact has not been fully elucidated. Present work was undertaken with a view to define the anatomical-physiological correlates underlying the role exercised by NPY in the regulation of GnRH in the forebrain of the teleost Clarias batrachus. Application of double immunocytochemistry revealed close associations as well as colocalizations of the two peptides in the olfactory receptor neurons (ORNs), olfactory nerve fibers and their terminals in the glomeruli, ganglion cells of nervus terminalis, medial olfactory tract, fibers in the area ventralis telencephali/pars supracommissuralis and cells as well as fibers in the pituitary. NPY containing axons were found to terminate in the vicinity of GnRH cells in the pituitary with light as well as electron microscopy. Double immunoelectron microscopy demonstrated gold particles for NPY and GnRH colocalized on the membrane and in dense core of the secretory granules in the cells distributed in all components of the pituitary gland. To assess the physiological implication of these observations, NPY was injected via the intracranial route and the response of GnRH immunoreactive system was evaluated by relative quantitative morphometry as well as high performance liquid chromatography (HPLC) analysis. Two hours following NPY (20 ng/g body weight) administration, a dramatic increase was observed in the GnRH immunoreactivity in the ORNs, in the fibers of the olfactory bulb (163%) and medial olfactory tract (351%). High performance liquid chromatography-electrospray ionization-mass spectrometric analysis confirmed the immunocytochemical data. Significant rise in the salmon GnRH (sGnRH)-like peptide content was observed in the olfactory organ (194.23%), olfactory bulb (146.64%), telencephalon+preoptic area (214.10%) and the pituitary (136.72%) of the NPY-treated fish. However, GnRH in the hypothalamus was below detection limit in the control as well as NPY-treated fish. Present results suggest the involvement of NPY in the up-regulation of sGnRH containing system at different level of neuraxis extending from the olfactory epithelium to the pituitary in the forebrain of C. batrachus. PMID- 15893650 TI - Expression of the glucocorticoid-induced receptor mRNA in rat brain. AB - The glucocorticoid-induced receptor (GIR) is an orphan G-protein-coupled receptor awaiting pharmacological characterization. GIR was originally identified in murine thymoma cells, and shows a widespread, yet not completely complementary distribution in mouse and human brain. Expression of the mouse GIR gene is modulated by dexamethasone in the brain and periphery, suggesting that GIR function is directly responsive to glucocorticoid signals. The rat GIR was cloned from rat prefrontal cortex by our group and was shown to be up-regulated following chronic amphetamine. The physiological role of GIR in the rat is not known at present. In order to gain a clearer understanding of the potential functions of GIR in the rat, we performed a detailed mapping of GIR mRNA expression in the rat brain. GIR mRNA showed widespread distribution in forebrain limbic and thalamic structures, and a more restricted distribution in hindbrain areas such as the spinal trigeminal nucleus and the median raphe nucleus. Areas with moderate to high levels of GIR include olfactory regions such as the nucleus of olfactory tract, hippocampus, various thalamic nuclei, cortical layers, and some hypothalamic nuclei. In comparison with previous studies, significant regional differences exist in GIR distribution in mouse and rat brain, particularly in the thalamus, striatum and in hippocampus at a cellular level. Overall, the expression of GIR in rat brain more closely approaches that seen previously in human than mouse, suggesting that rat models may be more informative for understanding the role of GIR in glucocorticoid physiology and glucocorticoid-related disease states. GIR mRNA distribution in the rat indicates a potential role of this receptor in the control of feeding and ingestive behavior, regulation of stress and emotional behavior, learning and memory, and, drug reinforcement and reward. PMID- 15893651 TI - Entrainment by a palatable meal induces food-anticipatory activity and c-Fos expression in reward-related areas of the brain. AB - Rats maintained under restricted feeding schedules (RFS) develop food anticipatory activity and entrainment of physiological parameters. Food entrainment is independent of the suprachiasmatic nucleus and depends on food entrainable oscillators (FEO). Restricted feeding schedules lead animals toward a catabolic state and to increase their food driven motivation, suggesting that in this process metabolic- and reward-related mechanisms are implicated. This study explored if motivation driven by a palatable meal is sufficient to produce food entrainment. To address this question, we evaluated whether daily fixed access to a highly palatable meal entrained (PME) locomotor activity, serum glucose and free fatty acids concentrations in rats maintained without food deprivation. The entrained response of PME rats was compared with rats entrained to RFS. In a second experiment, we used c-Fos-IR to identify structures in the central nervous system involved with PME. Rats showed anticipatory activity to a daily palatable meal, with a lower intensity than rats entrained to RFS. Anticipatory activity persisted at least for four cycles after interrupting palatable meal, suggesting that this persistence depends on an endogenous oscillator. Glucose and free fatty acids were not entrained in PME rats. c-Fos expression in limbic system nuclei was in phase with PME time, but not in the hypothalamus. Results suggest 1) that food deprivation, i.e. a catabolic state is not necessary for the expression of anticipatory activity; 2) that an increase in the motivational state due to taste and/or nutritional contents of palatable meal is sufficient to entrain behavior; and 3) that structures in the limbic system are involved in this entrainment process. The present study indicates that metabolic and motivational mechanisms are involved in food entrainment, and suggests that the FEO may be a multi oscillatory system distributed over different regulatory systems in the brain. PMID- 15893652 TI - Cannabinoids depress excitatory neurotransmission between the subthalamic nucleus and the globus pallidus. AB - The globus pallidus receives its major glutamatergic input from the subthalamic nucleus and subthalamic nucleus neurons synthesize CB1 cannabinoid receptors. The hypothesis of the present work was that CB1 receptors are localized in terminals of subthalamo-pallidal glutamatergic axons and that their activation leads to presynaptic modulation of neurotransmission between these axons and globus pallidus neurons. Patch-clamp studies were carried out on oblique-sagittal mouse brain slices. The subthalamic nucleus was stimulated electrically and the resulting excitatory postsynaptic currents (EPSCs) were recorded in globus pallidus neurons. The mixed CB1/CB2 receptor agonist R(+)-[2,3-dihydro-5-methyl-3 [(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-yl]-(1 naphthalenyl)methanone mesylate (WIN55212-2; 3 x 10(-7) M) had no effect on EPSCs. WIN55212-2 (10(-5) M) decreased the amplitude of EPSCs by 44+/-8%. The inhibition by WIN55212-2 (10(-5) M) was prevented by the CB1 antagonist N piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-3 pyrazolecarboxamide (10(-6) M). WIN55212-2 (10(-5) M) did not change the amplitude of spontaneous EPSCs (sEPSCs) recorded in globus pallidus neurons but lowered their frequency. Moreover, WIN55212-2 (10(-5) M) had no effect on currents elicited by direct activation of postsynaptic receptors on globus pallidus neurons by glutamate (10(-3) M) ejected from a pipette. In a final series of experiments, the firing of subthalamic nucleus neurons was recorded; WIN55212-2 (10(-5) M) did not change the firing of these neurons. The results show that activation of CB1 receptors inhibits glutamatergic neurotransmission between the subthalamic nucleus and the globus pallidus. Lack of effect of cannabinoids on the amplitude of sEPSCs and on currents evoked by direct stimulation of postsynaptic glutamate receptors indicates that the mechanism is presynaptic inhibition of glutamate release from axon terminals. Cannabinoids seem to act preferentially presynaptically: in contrast to their action on axon terminals, they have no effect on somadendritic receptors regulating firing rate. Cannabinoids elicit catalepsy in vivo. The observed inhibition of glutamatergic neurotransmission in the globus pallidus would favor catalepsy. PMID- 15893653 TI - Mapping the effects of the selective dopamine D2/D3 receptor agonist quinelorane using pharmacological magnetic resonance imaging. AB - Dopamine agonists with a high affinity for D2 and D3 receptors have a biphasic effect on rodent locomotion, inducing hypolocomotion at low doses and hyperlocomotion at higher doses. Controversy surrounds the role of the D3 receptor in mediating the hypolocomotor response to low agonist doses. This study examines patterns of neuronal activation induced by varying doses of the D2/D3 receptor agonist quinelorane using blood oxygen level dependent (BOLD) pharmacological magnetic resonance imaging (phMRI), and compares them with corresponding behavioural responses. Quinelorane (3 microg/kg) induced hypolocomotion in rats naive to the testing environment, and in phMRI experiments increased neuronal activity within the anterior olfactory nuclei, nucleus accumbens and islets of Calleja, regions containing a high density of D3 receptors. A 30 microg/kg dose of quinelorane resulted in biphasic locomotor effects, with initial hypolocomotion followed by sustained hyperlocomotion. phMRI indicated that this higher dose increased cerebral activity within limbic and olfactory regions, as did the lower drug dose, but induced additional activation in the caudate-putamen and globus pallidus, areas dense in D2 receptors but containing few D3 receptors. The more restricted pattern of activation at low agonist doses and close temporal relationship between behavioural and BOLD signal responses to quinelorane suggest that those nuclei most dense in D3 receptors play a key role in mediating the hypolocomotor effects of quinelorane. However, the presence of D3 receptors in activated brain regions may be coincidental, and further studies are required to show definitively which class of receptors mediates agonist-induced hypolocomotion. In contrast, the activation of D2 receptors within the striatum appears necessary for quinelorane-induced hyperlocomotion. PMID- 15893654 TI - Initiation and spread of epileptiform discharges in the methylazoxymethanol acetate rat model of cortical dysplasia: functional and structural connectivity between CA1 heterotopia and hippocampus/neocortex. AB - Neuronal migration disorders (NMDs) are often associated with medically intractable epilepsy. In utero injection of methylazoxymethanol acetate into pregnant rats gives rise to dysplastic cell clusters ("heterotopia") in hippocampus (and nearby regions), providing an animal model of NMD. In the present study, we have examined the structural and functional integration of hippocampal heterotopic cells into circuits that link the heterotopia with surrounding "normal" brain. Bi-directional morphological connectivity between the heterotopia and hippocampus/neocortex was demonstrated using the neurotracer, biotinylated dextran amine. Single cell recordings in hippocampal slices showed that heterotopia neurons form functional connections with the surrounding hippocampus and neocortex. However, simultaneous field recordings from the CA1 heterotopia, normotopic hippocampus, and neocortex indicated that epileptiform discharges (spontaneous events seen in slices bathed with high [K+]o and bicuculline) were rarely initiated in the heterotopia (although the heterotopia was capable of generating epileptiform discharges independently of normal brain regions). Further, in most of the experiments, the aberrant connectivity provided by CA1 heterotopia failed to function as a "bridge" for epileptiform discharges to propagate directly from low-threshold hippocampus to neocortex. These data do not support the hypothesis that NMDs (heterotopic cell populations) serve as a focus and/or trigger for epileptiform activity, and/or facilitate propagation of epileptiform events. PMID- 15893655 TI - 17-Beta estradiol rapidly enhances extracellular signal-regulated kinase 2 phosphorylation in the rat brain. AB - Physiological doses of 17-beta Estradiol (E2) rapidly induce mitogen-activated protein kinase (MAPK) phosphorylation in a variety of cell culture and tissue explant preparations. Rapid MAPK phosphorylation has been implicated as a critical step in estrogen's effects on neuronal activity, gene transcription and neuroprotection. The present series of in vivo experiments were designed to determine whether acute administration of estrogen rapidly increased extracellular signal-regulated protein kinase (ERK) 2 phosphorylation. Brains were harvested 20 min after a single i.p. injection of 15 microg/kg of 17-beta or 17-alpha estradiol. Twelve brain structures were micro-dissected, homogenized and processed for Western blotting. E2-treated rats exhibited a statistically significant increase in ERK2 phosphorylation in the diagonal band of Broca, rostral nucleus accumbens, paraventricular nucleus, arcuate nucleus and anteromedial visual cortex. Administration of the same dose of 17-alpha estradiol did not enhance ERK phosphorylation in any of the brain regions examined. The in vivo data presented here extend previously published in vitro data indicating that E2 rapidly activates MAPK in primary neuronal cultures, explants and cell lines. These data also indicate that MAPK activation is a potential mediator of estrogens effects in some but not all estrogen receptor containing regions of the brain. PMID- 15893656 TI - Transmembrane interactions of alpha/beta integrin signaling. PMID- 15893657 TI - Allosteric control of O2 reactivity in Rieske oxygenases. PMID- 15893658 TI - Reduce to increase: copper binding to a mitochondrial chaperone. PMID- 15893659 TI - Circular proteins: ring around with NOESY. PMID- 15893660 TI - Processing of a 22 kDa precursor protein to produce the circular protein tricyclon A. AB - Cyclotides are a family of plant proteins that have the unusual combination of head-to-tail backbone cyclization and a cystine knot motif. They are exceptionally stable and show resistance to most chemical, physical, and enzymatic treatments. The structure of tricyclon A, a previously unreported cyclotide, is described here. In this structure, a loop that is disordered in other cyclotides forms a beta sheet that protrudes from the globular core. This study indicates that the cyclotide fold is amenable to the introduction of a range of structural elements without affecting the cystine knot core of the protein, which is essential for the stability of the cyclotides. Tricyclon A does not possess a hydrophobic patch, typical of other cyclotides, and has minimal hemolytic activity, making it suitable for pharmaceutical applications. The 22 kDa precursor protein of tricyclon A was identified and provides clues to the processing of these fascinating miniproteins. PMID- 15893661 TI - A coiled-coil structure of the alphaIIbbeta3 integrin transmembrane and cytoplasmic domains in its resting state. AB - One of the hallmark features of the integrin receptors is the ability to transmit signals bidirectionally through the cell membrane. The transmembrane integrin domains are pivotal to the signaling events. An understanding of the signaling mechanism requires structural information. Here, we report a structural model of the transmembrane and part of the cytosolic domains of the alphaIIbbeta3 integrin in its resting state. The model was obtained computationally by a restrained conformational search of helix-helix interactions. It agrees with one published NMR structure of the cytoplasmic complex and can put many experimental findings on structural grounds. According to our model, integrins form an intricately designed coiled-coil structure in the resting state. The conserved Glycophorin A (GpA)-like sequence motif of the alpha, but not the beta, subunit, is in the interface of this model. Based on our calculations and other data, a signaling mechanism that involves a transient GpA-like structure is proposed. PMID- 15893662 TI - Folding studies of Cox17 reveal an important interplay of cysteine oxidation and copper binding. AB - Cox17 is a key mitochondrial copper chaperone involved in the assembly of cytochrome c oxidase (COX). The NMR solution structure of the oxidized apoCox17 isoform consists of a coiled-coil conformation stabilized by two disulfide bonds involving Cys(26)/Cys(57) and Cys(36)/Cys(47). This appears to be a conserved tertiary fold of a class of proteins, localized within the mitochondrial intermembrane space, that contain a twin Cys-x(9)-Cys sequence motif. An isomerization of one disulfide bond from Cys(26)/Cys(57) to Cys(24)/Cys(57) is required prior to Cu(I) binding to form the Cu(1)Cox17 complex. Upon further oxidation of the apo-protein, a form with three disulfide bonds is obtained. The reduction of all disulfide bonds provides a molten globule form that can convert to an additional conformer capable of binding up to four Cu(I) ions in a polycopper cluster. This form of the protein is oligomeric. These properties are framed within a complete model of mitochondrial import and COX assembly. PMID- 15893663 TI - SOMO (SOlution MOdeler) differences between X-Ray- and NMR-derived bead models suggest a role for side chain flexibility in protein hydrodynamics. AB - Reduced numbers of frictional/scattering centers are essential for tractable hydrodynamic and small-angle scattering data modeling. We present a method for generating medium-resolution models from the atomic coordinates of proteins, basically by using two nonoverlapping spheres of differing radii per residue. The computed rigid-body hydrodynamic parameters of BPTI, RNase A, and lysozyme models were compared with a large database of critically assessed experimental values. Overall, very good results were obtained, but significant discrepancies between X ray- and NMR-derived models were found. Interestingly, they could be accounted for by properly considering the extent to which highly mobile surface side chains differently affect translational/rotational properties. Models of larger structures, such as fibrinogen fragment D and citrate synthase, also produced consistent results. Foremost among this method's potential applications is the overall conformation and dynamics of modular/multidomain proteins and of supramolecular complexes. The possibility of merging data from high- and low resolution structures greatly expands its scope. PMID- 15893664 TI - Recent cyanobacterial Kai protein structures suggest a rotary clock. AB - The cyanobacterial circadian oscillator consists of three Kai proteins, KaiA, KaiB, and KaiC, in its oscillation feedback loop. Structural comparison reveals that the Kai system resembles the F1-ATPase system in which KaiC is equivalent to alpha(3)beta(3), KaiA to gammadelta, and KaiB to its inhibitory factor. It also suggests that there exists a possible haemagglutinin-like spring-loaded mechanism for the activation of KaiA during the formation of Kai complexes. PMID- 15893665 TI - The AXH domain adopts alternative folds the solution structure of HBP1 AXH. AB - AXH is a protein module identified in two unrelated families that comprise the transcriptional repressor HBP1 and ataxin-1 (ATX1), the protein responsible for spinocerebellar ataxia type-1 (SCA1). SCA1 is a neurodegenerative disorder associated with protein misfolding and formation of toxic intranuclear aggregates. We have solved the structure in solution of monomeric AXH from HBP1. The domain adopts a nonclassical permutation of an OB fold and binds nucleic acids, a function previously unidentified for this region of HBP1. Comparison of HBP1 AXH with the crystal structure of dimeric ATX1 AXH indicates that, despite the significant sequence homology, the two proteins have different topologies, suggesting that AXH has chameleon properties. We further demonstrate that HBP1 AXH remains monomeric, whereas the ATX1 dimer spontaneously aggregates and forms fibers. Our results describe an entirely novel, to our knowledge, example of a chameleon fold and suggest a link between these properties and the SCA1 pathogenesis. PMID- 15893666 TI - Scorpion-toxin mimics of CD4 in complex with human immunodeficiency virus gp120 crystal structures, molecular mimicry, and neutralization breadth. AB - The binding surface on CD4 for the HIV-1 gp120 envelope glycoprotein has been transplanted previously onto a scorpion-toxin scaffold. Here, we use X-ray crystallography to characterize atomic-level details of gp120 with this transplant, CD4M33. Despite known envelope flexibility, the conformation of gp120 induced by CD4M33 was so similar to that induced by CD4 that localized measures were required to distinguish ligand-induced differences from lattice variation. To investigate relationships between structure, function, and mimicry, an F23 analog of CD4M33 was devised. Structural and thermodynamic analyses showed F23 to be a better molecular mimic of CD4 than CD4M33. F23 also showed increased neutralization breadth, against diverse isolates of HIV-1, HIV-2, and SIVcpz. Our results lend insight into the stability of the CD4 bound conformation of gp120, define measures that quantify molecular mimicry as a function of evolutionary distance, and suggest how such evaluations might be useful in developing mimetic antagonists with increased neutralization breadth. PMID- 15893667 TI - The active conformation of the PAK1 kinase domain. AB - The p21-activated kinases (PAKs) participate in cytoskeletal control networks, downstream of Rho-family GTPases. A structure of PAK1 in an autoregulated, "off" state showed that a regulatory region, N-terminal to the kinase domain, forces the latter into an inactive conformation, prevents phosphorylation of Thr423 in the activation loop, and promotes dimerization. We have now determined structures at 1.8 A resolution for the free PAK1 kinase domain, with a mutation in the active site that blocks enzymatic activity, and for the same domain with a "phosphomimetic" mutation in the activation loop. The two very similar structures show that even in the absence of a phosphorylated Thr423, the kinase has an essentially active conformation. When Cdc42 binds the regulatory region and dissociates the dimer, PAK1 will be in an "intermediate-active" state, with a capacity to phosphorylate itself or other substrates even prior to modification of its activation loop. PMID- 15893668 TI - Crystal structure of a RuBisCO-like protein from the green sulfur bacterium Chlorobium tepidum. AB - Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzes the incorporation of atmospheric CO(2) into ribulose 1,5-bisphosphate (RuBP). RuBisCOs are classified into four forms based on sequence similarity: forms I, II and III are bona fide RuBisCOs; form IV, also called the RuBisCO-like protein (RLP), lacks several of the substrate binding and catalytic residues and does not catalyze RuBP-dependent CO(2) fixation in vitro. To contribute to understanding the function of RLPs, we determined the crystal structure of the RLP from Chlorobium tepidum. The overall structure of the RLP is similar to the structures of the three other forms of RuBisCO; however, the active site is distinct from those of bona fide RuBisCOs and suggests that the RLP is possibly capable of catalyzing enolization but not carboxylation. Bioinformatic analysis of the protein functional linkages suggests that this RLP coevolved with enzymes of the bacteriochlorophyll biosynthesis pathway and may be involved in processes related to photosynthesis. PMID- 15893669 TI - Implications for switching restriction enzyme specificities from the structure of BstYI bound to a BglII DNA sequence. AB - The type II restriction endonuclease BstYI recognizes the degenerate sequence 5' RGATCY-3' (where R = A/G and Y = C/T), which overlaps with both BamHI (GGATCC) and BglII (AGATCT), and thus raises the question of whether BstYI DNA recognition will be more BamHI-like or BglII-like. We present here the structure of BstYI bound to a cognate DNA sequence (AGATCT). We find the complex to be more BglII like with similarities mapping to DNA conformation, domain organization, and residues involved in catalysis. However, BstYI is unique in containing an extended arm subdomain, and the mechanism of DNA capture has both BglII-like and BamHI-like elements. Further, DNA recognition is more minimal than BglII and BamHI, where only two residues mediate recognition of the entire core sequence. Taken together, the structure reveals a mechanism of degenerate DNA recognition and offers insights into the possibilities and limitations in altering specificities of closely related restriction enzymes. PMID- 15893670 TI - Structural studies of the parainfluenza virus 5 hemagglutinin-neuraminidase tetramer in complex with its receptor, sialyllactose. AB - The paramyxovirus hemagglutinin-neuraminidase (HN) functions in virus attachment to cells, cleavage of sialic acid from oligosaccharides, and stimulating membrane fusion during virus entry into cells. The structural basis for these diverse functions remains to be fully understood. We report the crystal structures of the parainfluenza virus 5 (SV5) HN and its complexes with sialic acid, the inhibitor DANA, and the receptor sialyllactose. SV5 HN shares common structural features with HN of Newcastle disease virus (NDV) and human parainfluenza 3 (HPIV3), but unlike the previously determined HN structures, the SV5 HN forms a tetramer in solution, which is thought to be the physiological oligomer. The sialyllactose complex reveals intact receptor within the active site, but no major conformational changes in the protein. The SV5 HN structures do not support previously proposed models for HN action in membrane fusion and suggest alternative mechanisms by which HN may promote virus entry into cells. PMID- 15893671 TI - 2-Oxoquinoline 8-monooxygenase oxygenase component: active site modulation by Rieske-[2Fe-2S] center oxidation/reduction. AB - 2-Oxoquinoline 8-monooxygenase is a Rieske non-heme iron oxygenase that catalyzes the NADH-dependent oxidation of the N-heterocyclic aromatic compound 2 oxoquinoline to 8-hydroxy-2-oxoquinoline in the soil bacterium Pseudomonas putida 86. The crystal structure of the oxygenase component of 2-oxoquinoline 8 monooxygenase shows a ring-shaped, C3-symmetric arrangement in which the mononuclear Fe(II) ion active site of one monomer is at a distance of 13 A from the Rieske-[2Fe-2S] center of a second monomer. Structural analyses of oxidized, reduced, and substrate bound states reveal the molecular bases for a new function of Fe-S clusters. Reduction of the Rieske center modulates the mononuclear Fe through a chain of conformational changes across the subunit interface, resulting in the displacement of Fe and its histidine ligand away from the substrate binding site. This creates an additional coordination site at the mononuclear Fe(II) ion and can open a pathway for dioxygen to bind in the substrate containing active site. PMID- 15893672 TI - Structure of the angiopoietin-2 receptor binding domain and identification of surfaces involved in Tie2 recognition. AB - The angiopoietins comprise a small class of secreted glycoproteins that play crucial roles in the maturation and maintenance of the mammalian vascular and lymphatic systems. They exert their effects through a member of the tyrosine kinase receptor family, Tie2. Angiopoietin/Tie2 signaling is unique among tyrosine kinase receptor-ligand systems in that distinct angiopoietin ligands, although highly homologous, can function as agonists or antagonists in a context dependent manner. In an effort to understand this molecular dichotomy, we have crystallized and determined the 2.4 A crystal structure of the Angiopoietin-2 (Ang2) receptor binding region. The structure reveals a fibrinogen fold with a unique C-terminal P domain. Conservation analysis and structure-based mutagenesis identify a groove on the Ang2 molecular surface that mediates receptor recognition. PMID- 15893675 TI - Epidemiology and etiology of multiple sclerosis. AB - The author believes that the Faroese saga provides major insight into what seems to him to be the essential nature of MS: There is a specific, widespread, but unidentified, infection that we call the primary multiple sclerosis affection (PMSA). PMSA is a persistent infection that is transmitted from person to person. A small proportion of persons who has PMSA will develop clinical neurologic multiple sclerosis (CNMS) years later. Prolonged exposure is needed to acquire PMSA. Acquisition follows first adequate exposure. Susceptibility to PMSA is limited to approximately age 11 to age 45 at start of exposure. CNMS is not transmissible.PMSA transmissibility is limited to a period that is less than the usual age of onset of CNMS. On the Faroe Islands, this period is approximately from age 13 to age 26. The existence of PMSA now must be inferred from the presence of CNMS. PMID- 15893676 TI - Multiple sclerosis: an immunologic perspective. AB - The precise cause of MS remains unknown, although immunopathologic studies provide solid evidence of participation of activated immune cells in the formation of MS lesions. Cell-mediated and antibody-mediated(humoral) immune responses are involved in the immunopathogenesis of MS. The Thl and Th2 cells, by releasing a variety of pro- or anti-inflammatory cytokines, work in opposition to balance the immune response and to determine the net effect of the inflammation. The current understanding of immunopathogenesis shapes the contemporary approach to the treatment of MS. The article also reviewed the mechanisms of action of commonly used immunotherapy agents and their impact on the immune response of patients who have MS. PMID- 15893677 TI - Diagnosing multiple sclerosis and its imitators. AB - The diagnosis of MS requires central nervous system symptoms that are disseminated in time and space, and that have no better explanation. Dissemination in time and space may be demonstrated clinically or by MRI imaging. The differential diagnosis is broad, and requires the exclusion of several diseases that are described in the text. Following the new guidelines for the diagnosis of MS allows an early and accurate diagnosis of the disease. PMID- 15893678 TI - MRI in multiple sclerosis. AB - MRI provides multiple uses and applications in multiple sclerosis(MS). The basic features of the MRI-detected lesions, including the underlying pathology, are discussed. MRI allows assessment of the normal-appearing white and gray matter, and neuronal tract and functional system disturbances. An overview of the clinical significance of these MRI measures is included, as a basis for understanding their role as outcome measures in clinical trials. MRI recently assumed greater importance in its role in establishing an earlier diagnosis of MS after a first clinical event, and in monitoring subclinical disease before or subsequent to the formal diagnosis. The background to these applications and practical issues are discussed. PMID- 15893679 TI - Neuropsychological evaluation and treatment of multiple sclerosis: the importance of a neuro-rehabilitation focus. AB - This article describes the components of a neuropsychologic evaluation and some of the primary indications for its use in multiple sclerosis (MS). We also detail the kinds of neurocognitive and neurobehavioral problems that are cited commonly in the relevant literature and seen in the clinical setting. We provide a brief overview of the brain structures that are affected commonly by MS, and their implications for neuropsychologic function. We have included an overview of some of the current medications that are used to target cognitive and emotional symptoms that can be a direct result of the disease. We also present four representative case examples of composite patients, and briefly review the ways in which neuropsychologic evaluation and neuro-rehabilitation treatments can help people who have MS. PMID- 15893680 TI - Depression in persons with multiple sclerosis. AB - This article provides a critical review of what is currently known concerning the nature and scope of depression in persons who have MS. It begins with a discussion of relevant terminology,which is followed by a summary of the research literature concern-ing the prevalence; severity; and risk factors for, and impact of, depression in MS. Assessment issues also are reviewed. Gaps in the literature on the treatment of depression in persons who have MS also are discussed. The article concludes with a discussion of future directions that will improve our understanding of depression, its treatment, and the other side-resilience in the face of a serious neurologic condition. PMID- 15893681 TI - A practical approach to immunomodulatory therapy for multiple sclerosis. AB - This article provides a current overview of the definition and pathogenesis of the disease, the different types of MS, a new diagnostic criteria, the rationale for early therapy, a review of the approved MS therapies, the strategies to evaluate ongoing treatment efficacy,the management of suboptimal treatment responders, and the prospects for future therapies. The article focuses on relapsing remitting MS because most of the therapeutic data deal with this type of MS. The role of mitoxantrone as a "rescue therapy" for suboptimal responders to IMTs is discussed. PMID- 15893682 TI - Multiple sclerosis and spasticity. AB - Spasticity is a common impairment in MS. It can result in significant medical complications and is associated with increased disability. Treatment strategies include skilled rehabilitation strategies, neuromuscular blocks, oral agents, intrathecal management, and surgery. Rehabilitation strategies are central, whereas other strategies are added based on the level of impairment and functional loss. Treatment strategies for spasticity management are far from optimal and are complicated in MS as a result of lesions in the brain and the spinal cord. Pharmaceutical management in MS is complicated by the numerous secondary impairments in MS and its associated polypharmacy.Head-to-head studies of the various agents are rare. The studies that exist are small and do not point to any one strategy over another. Although management is difficult, it is essential for the health, functional status, and well-being of the individual who has MS. Providers must use well-developed clinical skills to arrive at optimal individualized treatment programs and monitor them frequently. For spasticity that is unresponsive, referral to a MS Center with a spasticity program is ideal. PMID- 15893683 TI - Multiple sclerosis-related fatigue. AB - Fatigue is a significant factor in the lives of many MS patients and the most commonly reported symptom in many studies. Fatigue is an important symptom to consider because it affects patients' social lives, occupations, and activities of daily living. Efforts to predict fatigue have been mixed, but it appears to be related to overall quality of life and mood. From a pathophysiologic perspective, fatigue in MS is multifactorial and complex,involving dysregulation of the immune system, changes in the nervous system related to the disease process, neuroendocrine and neurotransmitter changes, and other factors such as physical deconditioning, sleep disturbance, pain, and medication side effects. Various attempts to assess fatigue have been made, and many measures are now available for use in clinical practice and research. In clinical practice, these measures help guide treatment considerations. Recent research has provided valuable strategies to ameliorate fatigue in MS, and although many patients continue to experience fatigue despite interventions, many receive substantial relief.Nonpharmacologic approaches-considered the first step in treatment-include exercise programs, cooling, dietary considerations, and energy conservation strategies. For patients who continue to experience significant fatigue, several medications (although not specifically approved for use in the reduction of MS related fatigue) have proved effective in this regard.The first-line agents include amantadine for mild fatigue and modafinil for more severe cases. Second line agents include pemoline and antidepressant medications. Other pharmacologic agents have also shown some promise. PMID- 15893684 TI - Chronic pain in persons with multiple sclerosis. AB - The MS literature clearly indicates that chronic pain is a significant problem for many, although not all, persons with MS. The rates of pain have been found to vary in different studies, from 44% to 80%, depending on the sample and the specific questions used to assess the incidence and severity of pain. What is not clear is the proportion of persons who have acute pain relative to chronic pain. Although the specific frequency of pain problems in patients with MS may not be clear, there is a subgroup of patients (about 38% of those with pain in one sample) who report experiencing severe pain [8]. Preliminary research suggests that chronic pain can have a significant negative impact on a number of aspects of functioning in persons with MS, such as the ability to engage in household work and psychologic functioning. A biopsychosocial model of chronic pain, which has proved to be useful in understanding chronic pain as a primary condition and chronic pain in persons with other physical disabilities, may also be useful for understanding pain in persons with MS. Research,however, has not yet tested the utility of this model among MS populations. Longitudinal research is needed to help us learn how MS-related pain may fluctuate over time and with changes in disease status. There also is a strong need for research that examines access to pain treatment and that evaluates the efficacy of currently available pain treatments in persons with MS. The results of such research, as it is applied to help patients with MS, should contribute to an overall increase in well-being and a decrease in suffering among persons with MS and chronic pain. PMID- 15893685 TI - Exercise and rehabilitation for individuals with multiple sclerosis. AB - It is the coexistence of physical and cognitive impairments, together with emotional and social issues in a disease with an uncertain course, that makes MS rehabilitation unique and challenging. Inpatient rehabilitation improves functional independence but has only limited success improving the level of neurologic impairment. Benefits are usually not long lasting. Severely disabled people derive equal or more benefit than those who are less disabled, but cognitive problems and ataxia tend to be refractory. There is now good evidence that exercise can improve fitness and function for those with mild MS and helps to maintain function for those with moderate to severe disability. Therapy can be performed over 6 to 15 weeks in outpatient or home-based settings or as a weekly day program lasting several months. Several different forms of exercise have been investigated. For most individuals, aerobic exercise that incorporates a degree of balance training and socialization is recommended. Time constraints, access, impairment level, personal preferences, motivations, and funding sources influence the prescription for exercise and other components of rehabilitation. Just as immunomodulatory drugs must be taken on a continual basis and be adjusted as the disease progresses, so should rehabilitation be viewed as an ongoing process to maintain and restore maximum function and QOL. PMID- 15893686 TI - Health promotion in people with multiple sclerosis. AB - Chronic diseases like MS present unique challenges and opportunities for patients and the medical care system. Patients are challenged because they are under tremendous pressure to actively engage themselves in multiple prevention, treatment, and health maintenance behaviors, often before they feel ready. Health care providers are challenged because health-promotion activities require more time, counseling skills, and organizational resources than traditional, acute medical care. Patients, clinicians, and researchers face the challenge of determining which health-promotion activities are not only supported by the evidence but also appropriate for a given patient. New models of health promotion are being developed that integrate self-help and professional help. These approaches have been applied in other chronic diseases and should be adapted and studied among people with MS. PMID- 15893687 TI - Mitigating the impact of multiple sclerosis on employment. AB - This article reviews the employment status of people with MS, the importance of employment, key barriers to employment, and compensatory and accommodation strategies to enhance employment. Resources and recommendations for health care providers to assist their patients in maintaining, securing, or leaving employment are presented. PMID- 15893688 TI - Taking part in life: enhancing participation in multiple sclerosis. AB - Health care professionals participating in rehabilitation for people with MS can play a critical role in enhancing limited outcomes such as enhanced mobility, reductions in symptoms such as pain and depression, and the metaoutcome participation. This role will be significantly more effective if the health care professional acknowledges and validates the different perspectives of the professional and the patient and recognizes the expertise of the patient who has lived with MS in the context of his or her life. Assuming this role effectively requires that the health care professional develop a collaborative relationship with the patient and understand that the role may change depending on the stage of MS and the individual's circumstances. PMID- 15893689 TI - Novel biological networks modulated by complement. AB - The almost complete deciphering of the human genome has paved the way for the application of new technology platforms in understanding the contribution of complex biological pathways to human pathophysiology and disease. In the post genomic era, the concept of systems biology has gained significant momentum and biomedical research is now being conducted on an integrated and cross disciplinary platform that pulls together its resources from diverse fields such as computational biology, bioinformatics, functional genomics, structural biology, and proteomics. In this perspective, the identity of established biologic systems is being re-examined in the light of novel findings that suggest novel associations between otherwise unrelated pathways and individual proteins. Complement exemplifies such a system that, transcending its innate immune identity, has forged functional associations with multiple pathways and networks in modulating basic biologic processes. In the present article, we provide a global overview of these unusual system associations of complement with the aid of a powerful and high-throughput bioinformatics platform. Using a novel approach called systems literature analysis that allows the rapid extraction of text-based associations between genes and pathways from the ever expanding scientific article database, we have selected a broad range of biologic processes modulated by complement proteins and have constructed an integrated map of complement mediated networks that incorporates well over 85 diverse biologic pathways. Expanding the complement cascade beyond its approximately 35 designated components, we discuss protein-protein interactions involving novel ligands and associations with signaling cascades and cellular networks that affect both inflammatory and non-inflammatory processes. This integrated consideration of complement within a unified 'systems biology' framework underscores the concept that innate immunity goes well beyond the protection of 'self' extending links to critical developmental, homeostatic, and metabolic processes. PMID- 15893690 TI - Seeing the light with celiac. PMID- 15893691 TI - Combinational IL-2/IL-15 induction does not further enhance IL-15-induced lymphokine-activated killer cell cytotoxicity against human leukemia/lymphoma cells. AB - In vitro comparisons of induction of perforin (PFP), granzyme B (GRB), production of cytokines, and cell-mediated cytotoxicity by interleukin-2 (IL-2), interleukin 15 (IL-15), or combinational IL-2/IL-15-induced lymphokine-activated killer cells were studied in this study. Whereas IL-2-induction was associated with a decrease in cultured cell population over a 7-day period, IL-15 alone or in combination with IL-2 resulted in significant increase including cytotoxic T lymphocytes and subsets of CD56+ lymphocytes, particularly cytokine-induced killer and cytolytic natural killer-T lymphocytes. The overall PFP, GRB, and tumor necrosis factor alpha expression in different subtypes were also significantly higher with IL-15 alone or in combination with IL-2 induction with resultant superior cytotoxicity compared to IL-2 treatment. There was no significant advantage of addition of IL 2 over IL-15 induction. These results offer further information on the cytotoxic potency of these cytokines and their mechanisms of action implicating potential use of IL-15 as part of cytokine adoptive immunotherapy. PMID- 15893692 TI - Infliximab induces apoptosis of monocytes and T lymphocytes in a human-mouse chimeric model. AB - Tumor necrosis factor (TNF) antagonism with monoclonal antibodies is an effective therapy for severe Crohn's disease and rheumatoid arthritis. Recent studies have suggested that induction of apoptosis of inflammatory cells contributes to this therapeutic effect. We investigated whether infliximab (a mouse-human IgG1 chimeric anti-TNF monoclonal antibody) could induce apoptosis in vivo in human mouse chimeras, created by reconstitution of severe combined immunodeficiency/beige mice with THP-1 (human monocytic cell line) or Jurkat cells (human T cell line). Infliximab treatment of chimeric mice depleted spleen and peritoneum from THP-1 cells and Jurkat cells and decreased production of the human cytokines IL-10 and IL-12 in vivo. Cell death was shown to occur already within 1 h of treatment. Infliximab effects were independent of FcgammaR binding or complement activation. Cell death resulted from apoptosis induction in a caspase-dependent pathway, as evidenced by the in vitro protective effect of the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethyketone (Z-VAD FMK). These data provide support for caspase-dependent apoptosis induction being the mechanism of action of infliximab in vivo. PMID- 15893693 TI - IA-2 combined epitope assay: a new, highly sensitive approach to evaluate IA-2 humoral autoimmunity in type 1 diabetes. AB - Islet tyrosine phosphatase 2 (IA-2) is one of the major autoantigens in type 1 diabetes. The aim of this work was to evaluate which IA-2 construct(s) among those usually employed has the highest sensitivity and specificity for detecting IA-2 autoantibodies in autoimmune diabetes and whether the combination of different IA-2 constructs into a single assay allows the detection of immunoreactivities otherwise not detectable by a single construct. For this purpose, we tested the single immunoreactivities of IA-2 FL(aa 1-979), IA 2(BDC)(aa 256-556:630-979), IA-2 IC(aa 605-979), IA-2(aa 256-760), IA-2(aa 761 928), and of 7 combinations of these fragments in the sera of 203 newly diagnosed type 1 diabetic patient (DM: 109 males,94 females, mean age 12.9 +/- 7.5 years) and 43 prediabetic subject (PDM: 20 males, 23 females, mean age 10.3 +/- 6.0 years) sera. IA-2 IC was the single construct that showed the highest sensitivity and specificity both in DM and PDM subjects; however, all of the other IA-2 constructs investigated detected additional immunoreactivities with respect to it. The combined use into the same assay of IA-2 IC, IA-2 FL, and IA-2 (256-760) constructs allowed detection of IA-2 Abs in additional 13.3% DM and 30.4% PDM subjects compared to the single IA-2 IC construct, suggesting this methodology as a new, highly sensitive approach to the study of IA-2 autoimmunity in type 1 diabetes. PMID- 15893694 TI - IL-17R activation of human airway smooth muscle cells induces CXCL-8 production via a transcriptional-dependent mechanism. AB - Airway neutrophilia has been recognized as a predominant feature of acute lung disorders. While it has been shown that IL-17 induces expression of the CXC chemokines in the airways leading to neutrophil recruitment, the IL-17R expression in human ASM cells and the molecular mechanism by which IL-17 mediates neutrophilic chemo-attractant CXCL-8 (IL-8) production have not been determined. Our study showed that ASM cells express steady state IL-17R protein, mRNA and surface-bound receptor. Interestingly, airway sections from COPD patients revealed IL-17R-positive immunostaining within ASM bundles. IL-17 was capable of stimulating CXCL-8 protein release from ASM cells which was significantly decreased by neutralizing anti-IL-17 mAb. Furthermore, IL-17 induction of CXCL-8 mRNA and protein release from ASM cells was abrogated by transcriptional inhibitor actinomycin D. CXCL-8 promoter reporter analysis using wild type and site specific mutant constructs demonstrated a key role for AP1 and NF-kappaB binding sites in IL-17-induced CXCL-8 expression. These data demonstrate that IL 17 mediates CXCL-8 expression in ASM cells via a transcriptional mechanism depending on NF-kappaB and AP-1 pathways. Together, our findings suggest that ASM cells play an important role in airway neutrophilia. PMID- 15893695 TI - Analysis of class switch recombination and somatic hypermutation in patients affected with autosomal dominant hyper-IgM syndrome type 2. AB - Autosomal recessive form of hyper-IgM syndrome type 2 (AR-HIGM2) is secondary to mutations affecting both alleles of AICDA gene encoding activation-induced cytidine deaminase, characterized by defects of immunoglobulin class switch recombination (CSR) and somatic hypermutation (SHM) in most of the patients. We herein report the immunological phenotype of seven patients carrying a single heterozygous R190X mutation in AICDA. Variable defect in in vivo CSR inherited as an autosomal dominant (AD) trait strongly suggests that this heterozygous AICDA mutation causes HIGM (AD-HIGM2). In AD-HIGM2 B cells, CSR was consistently found impaired in vitro. However, in contrast to AR-HIGM2, the CSR-induced double stranded DNA breaks in the switch region of IgM heavy chain gene were detected. The SHM frequency in V regions of IgM heavy chain gene in B cells was normal in all (but one patient). The characteristics of the AD-HIGM2 phenotype indicate that the AID C-terminal region may be involved in DNA repair machinery required for CSR. PMID- 15893696 TI - Increased expression of soluble decoy receptor 3 in acutely inflamed intestinal epithelia. AB - Decoy receptor 3 (DcR3), a soluble receptor in the tumor necrosis factor (TNF) receptor family, is known to inhibit apoptosis mediated by pro-apoptotic TNF family cytokines such as Fas ligand (FasL), TL1A, and LIGHT. Therefore, the regulation of DcR3 expression under certain pathophysiological conditions is of interest since the level of soluble DcR3 would most likely affect the homeostasis of cells and tissues. We found that human intestinal epithelial cell (IEC) lines (SW480, SW620, and HT29) could selectively increase DcR3 release in response to lipopolysaccharide (LPS) and that all the cells preferentially expressed Toll like receptor 4 (TLR-4). LPS-induced DcR3 releases in IECs appeared to be via the activation of mitogen-activated protein kinases (MAPK) such as extracellular signal-regulated kinase 1 and 2 (ERK1/2) and c-Jun NH2-terminal protein kinase (JNK), and the transcription factor NF-kappaB. Moreover, the increased expression of DcR3 in appendix epithelia from patients with acute appendicitis was demonstrated. Taken together, the results indicated that DcR3 might play an important role in the human intestinal epithelium during acute inflammatory processes caused by endotoxin challenge. PMID- 15893697 TI - Integration of high-risk human papillomavirus DNA correlates with HLA genotype aberration and reduced HLA class I molecule expression in human cervical carcinoma. AB - In human cervical cancer (CC), local immunity against this human papilloma virus (HPV)-associated neoplasia has been signified. To stratify the possibility of HPV integration on HLA mutations, we measured the genotypic and phenotypic integrity of all available HLA class I loci in 30 cases of CC. Paired normal and cancer genomic DNA was analyzed with DNA typing trays, including 57 subtypes of HLA-A, 120 subtypes of HLA-B, and 60 subtypes of HLA-C. We demonstrated significant mutations of HLA genotype with reduced HLA molecule expression in CC. HPV coincide in > 70% cases of aberrant HLA genes. Southern blot analysis revealed the presence of HPV DNA within the mutated HLA foci. Our study reveals a plausible role of HPV integration in the contexts of aberrant HLA genotypes in CC cells. Disruptions of the HLA genes can be possible tactics of HPV to attain the potential carcinogenetic purposes, and thus the cancer immune escape. PMID- 15893698 TI - Comparative potency of Ara h 1 and Ara h 2 in immunochemical and functional assays of allergenicity. AB - To assess the relative potency of the major peanut allergens, Ara h 1 and Ara h 2, we examined the relative ability of purified proteins to bind IgE on immunoblots, to cross-link allergen specific IgE in an in vitro assay of degranulation based on RBL SX-38 cells, and to bind IgE in the ImmunoCap assay. Sera from 12 highly sensitive, peanut allergic patients were studied in all assays. IgE immunoblots with crude peanut extracts showed binding of IgE to multiple bands including the 63 kDa and 17-19 kDa bands that contain Ara h 1 and Ara h 2, respectively. In the functional assay, Ara h 2 was more potent than Ara h 1 in 11 of 12 sera tested with a median potency that was 52.5-fold more than Ara h 1 (P < 0.005). Contrary to findings with the functional assay, IgE immunoblots with purified Ara h 1 and Ara h 2 showed substantially lighter binding of IgE to Ara h 2 compared with Ara h 1 (P = 0.02). The ImmunoCap assay gave intermediate results with slightly more IgE binding to Ara h 2 than to Ara h 1 (P = 0.005). In conclusion, Ara h 2 is a very potent allergen and is much more potent than Ara h 1 for most sera using an in vitro assay of IgE cross-linking and cell activation. This finding is different from what was predicted based on immunoblots or with the ImmunoCap assay. PMID- 15893699 TI - Characterization of in vivo expanded OspA-specific human T-cell clones. AB - A panel of CD4 T-cell clones was isolated from synovial fluid by single cell flow cytometry from a patient with treatment-resistant Lyme arthritis using a DRB1*0401 major histocompatibility complex (MHC) class II tetramer covalently loaded with outer surface protein A (OspA) peptide164-175, an immunodominant epitope of Borrelia burgdorferi. Sequencing of the T-cell receptors of the OspA reactive clones showed significant skewing of the T-cell receptor repertoire. Of the 101 T-cell clones sequenced, 81 possessed TCR beta chains that were present in at least one other clone isolated. Complete sequencing of both alpha and beta chains of a subset of clones showed that at least two distinct T-cell clones were expanded in vivo. Binding studies using a panel of Ala-substituted peptide ligands were performed to determine potential MHC binding sites of the OspAp164 175 to DRB1*0401. In addition, T-cell clones were tested functionally for their reactivity to the wild-type peptide as well as to altered peptide ligands (APLs) and peptide libraries based on the OspA epitope in order to determine the TCR contact residues and the stringency in T cell recognition. We are among the first to define the characteristics of TCR usage of T cells isolated from an inflamed immune compartment in an individual with an autoimmune disease potentially triggered by a microbial antigen. PMID- 15893700 TI - Morphine modulates chemokine gene regulation in normal human astrocytes. AB - Chemokines and their receptors have been implicated in the pathogenesis of neuroAIDS. Herein we describe the effects of morphine on the gene expression of beta chemokines and their receptors by primary normal human astrocytes (NHA). Our results show that NHA treated with morphine showed significant downregulation of the gene expression of beta chemokines, MCP-1, and MIP-1 beta, while reciprocally upregulating the expression of their specific receptors, CCR2b, CCR3, and CCR5 as detected by real-time quantitative PCR. These morphine-induced effects on NHA cells were reversed by the opioid mu receptor antagonist, naloxone. Further, our results indicate that morphine-induced effects are mediated via the modulation of MAPK and CREB signaling pathways. These results support our hypothesis that opiates act as co-factors in the neuropathogenesis of HIV infection. PMID- 15893701 TI - SARM1: A candidate gene in the onset of hereditary infectious/inflammatory diseases. PMID- 15893702 TI - Gastric electrical stimulation modulates neuronal activity in nucleus tractus solitarii in rats. AB - Implantable gastric electric stimulation (GES) has been under investigation for the treatment of gastric motor disorders and obesity. However, possible central mechanisms involving the effects of GES on gastric function are unclear. The purpose of this study was to examine the effects of GES with different parameters on neuronal activity in the nucleus tractus solitarii (NTS) of the medulla. Extracellular potentials of single neurons in NTS were recorded in pentobarbital anesthetized, paralyzed, ventilated male rats. GES with four sets of parameters was applied for one minute: GES-A (6 mA, 0.3 ms, 40 Hz, 2 s-on and 3 s-off), GES B (20 mA, 0.3 ms, 40 Hz, 2 s-on and 3 s-off), GES-C (6 mA, 6 ms, 40 Hz, 2 s-on and 3 s-off), and GES-D (6 mA, 200 ms, 12 imps/min). 35/118 (30%) neurons in NTS were responsive to gastric distension (GD, 20 mmHg, 20 s). Forty-one percent, 67%, 76% and 42% of all the responsive NTS neurons were affected by GES-A, -B, -C and -D, respectively. More NTS neurons with gastric inputs were affected with GES C (19/25) than with GES-A (11/27, P<0.05) and GES-D (10/24, P<0.05). Maximal excitatory responses (17.9+/-2.6 imp/s) of NTS neurons to GES-C were significantly greater than GES-D (9.7+/-4.8 imp/s, P<0.05), whereas average duration of excitatory response (74.8+/-4.3 s) of NTS neurons to GES-B was significant longer than GES-A (60.3+/-3.3 s). Gastric electrical stimulation primarily has an excitatory effect on NTS neurons receiving input from the stomach; the central neuronal response to GES is enhanced with stimulation using an increased pulse width and/or amplitude. This modulatory effect of GES on the central neurons receiving vagal inputs may contribute to the neural mechanisms of GES therapy for the treatment of patients with obesity and gastric motility disorders. PMID- 15893703 TI - Ventricular reflex interactions during transmural myocardial ischemia. AB - The role of left ventricular receptors with sympathetic afferent fibers in the reflex response to myocardial ischemia is controversial, particularly in the canine model. Previous experiments have shown that reflex excitatory responses mediated by left ventricular sympathetic afferents can be detected in sinoaortic denervated and vagotomized dogs during transmural myocardial ischemia. The purpose of these experiments was to determine if reflex excitatory responses occur in dogs with intact left ventricular vagal afferents. Experiments were performed in 27 chloralose-anesthetized dogs with sinoaortic denervation. Responses of efferent renal sympathetic nerve activity, arterial, and left atrial pressures to transmural and non-transmural inferoposterior myocardial ischemia were measured before and after interruption of left ventricular sympathetic afferents by stellectomy. The adequacy of sympathetic deafferentation was assessed by measurement of renal nerve responses to epicardial bradykinin. Prior to stellectomy, excitatory responses were observed in 10 animals and inhibitory responses in 9 animals. The remaining animals had no responses and were excluded from analysis. In the excitatory group, reflex increases in renal nerve activity during both transmural and non-transmural inferoposterior ischemia were abolished by stellectomy and not replaced by inhibitory responses. In the inhibitory group, non-transmural inferoposterior ischemia elicited greater reflex decreases in renal nerve activity when left ventricular sympathetic afferents were intact. After stellectomy, transmural ischemia elicited greater reflex inhibition of renal nerve activity. Renal nerve responses to epicardial bradykinin were abolished by stellectomy. These results indicate that reflex excitatory responses mediated by left ventricular receptors with sympathetic afferent fibers can be elicited in dogs with intact vagal afferents. These excitatory responses are most apparent during transmural myocardial ischemia. In dogs with inhibitory responses to coronary occlusion, activation of sympathetic afferents during transmural ischemia appears to attenuate reflex inhibitory responses mediated by left ventricular vagal afferents. PMID- 15893704 TI - A neurophysiological evidence of capsaicin-sensitive nerve components innervating interscapular brown adipose tissue. AB - Neurophysiological basis for the heterogeneity of the nerve components in the brown adipose tissue (BAT) was examined in this experiment. Efferent nerve signals were recorded from the central cut end of the small nerve filament dissected from the nerve fibers innervating the interscapular BAT (IBAT). By focusing on qualitative aspects of observed compound action potentials (spikes), we found two distinctive types of spikes exhibited by the intercostal nerves innervating IBAT. The spikes mainly appeared upon sympathetic stimulations (cold stimulation and glucose administration) were characterized by low amplitude with relatively short duration (small spike) and their sensitivity to the ganglion blocker, hexamethonium (C6). On the other hand, the spikes seen throughout the experiments were characterized by high amplitude with long duration (large spike) and their insensitivity to C6. Since BAT is activated by cold and feeding via sympathetic nervous system, the small spikes seemed to be exhibited by sympathetic fibers. On the other hand, appearance of the large C6-insensitive spikes was strongly attenuated in capsaicin-desensitized rats. Even though the functional link between IBAT and C6 insensitive fibers remains unanswered, our results suggest that IBAT is under control of various nerve types including capsaicin-sensitive fibers in addition to the control of sympathetic nervous system. PMID- 15893705 TI - Different modulation of spontaneous activities by nitrergic inhibitory nerves between ileum and jejunum in W/Wv mutant mice. AB - We compared the spontaneous electrical and mechanical activities between the jejunum and ileum in the W/Wv mutant mouse, where ICC in the myenteric region (ICC-MY) are deficient. Electrical slow waves (SWs) superimposed with spike potentials, and synchronous circular and longitudinal muscle mechanical activities at a regular rhythm under approximately 1 cm H2O were recorded in the jejunum and ileum of wild-type mice. However, in the jejunum and ileum of W/Wv mice, irregular electrical and mechanical activities without discernable SWs were recorded. N-nitro-L-arginine methyl ester (L-NAME) significantly decreased the mean interval of longitudinal muscle contractions from 4.43+/-3.39 to 2.50+/-1.23 s in the ileum of W/W(V) mice. L-NAME also significantly decreased mean coefficient of variance (decreased irregularity) in the intervals from 2.59+/ 0.84 to 0.48+/-0.46 in the ileum. Tetrodotoxin also significantly decreased mean interval and coefficient of variance in the ileum. Neither L-NAME nor tetrodotoxin affected contractile activity in the jejunum. These results suggest that enteric nitrergic nerves in the ileum, but not the jejunum, mediate a steady state inhibition of myogenic activity in W/Wv mice. PMID- 15893706 TI - Short-term synaptic plasticity in rabbit pancreatic ganglia. AB - The extrinsic innervation of the pancreas converges on a plexus of intrinsic pancreatic ganglia whose cholinergic neurons innervate acini, ducts, islets and blood vessels. Therefore, understanding ganglionic transmission is essential for understanding neural control of pancreatic secretion. Intracellular recordings of nicotinic fast excitatory postsynaptic potentials (fEPSPs) and action potentials (APs) were used to characterize and compare transmission in ganglia from the head/neck and body regions of the rabbit pancreas. Paired-pulse facilitation (PPF) or depression (PPD) of fEPSPs was observed in ganglia from both regions with PPF peaking and disappearing at shorter inter-stimulus intervals than PPD. PPF was most frequent in the head/neck (60%) and PPD (50%) in the body. Repetitive stimulation (10 Hz/5 s) evoked multiple forms of mid- and post-train plasticity. Facilitation during the first 1-2 s of train stimulation was reduced or reversed with continued stimulation due to development of synaptic depression and mid-train depression was of greater magnitude in the head/neck region. A brief (approximately 10 s) post-train augmentation was followed by a 1-2 min post train depression that appeared to result from inhibition of ACh release. Regional differences in the frequency, magnitude, or duration of all forms of synaptic plasticity suggested regional differences in the extrinsic innervation patterns and possibly the function of pancreatic ganglia. In conclusion, rabbit pancreatic ganglia exhibit multiple forms of short-term synaptic plasticity that markedly alter the probability of postsynaptic firing, consistent with these ganglia being critical sites of synaptic integration and autonomic regulation of pancreatic secretion. PMID- 15893707 TI - Effects of body posture on local sweating and sudomotor outflow as estimated using sweat expulsion. AB - To estimate the effects of changes in body posture on sudomotor function, sweat rates on the forearm, chest and thigh, tympanic temperature (Tty), and skin temperatures were recorded in an upright sitting and a supine position under a hot environment of 40 degrees C Ta and 40% relative humidity for 60 min. Sweat expulsions were identified on sweat rate curves and their rates (Fsw) were calculated. Tty was higher, and its initial fall was greater, in the supine position than in the sitting position. On the forearm and the chest, the regression line relating sweat rate to mean body temperature (Tmb) had a gentler slope in the supine position, whereas on the thigh, it showed a steeper slope. The regression line relating Fsw to Tmb had a steeper slope in the supine position than in the sitting position, suggesting that the gain in the mechanisms for central integration and rhythm-generation was enhanced in the supine position. The parameter of sweat rate divided by Fsw was lower on the forearm and the chest, whereas it was higher on the thigh in the supine position than in the sitting position, suggesting that sudomotor outflow was modified at the spinal cord in association with skin pressure. It was concluded that body posture affects sudomotor functions through both brain and spinal mechanisms. PMID- 15893708 TI - Metaiodobenzylguanidine (MIBG) scintigraphy at various parts of the body in Parkinson's disease and multiple system atrophy. AB - We compared MIBG uptake at various parts of the body in controls and patients with Parkinson's disease and multiple system atrophy. In the heart, MIBG uptake in Parkinson's disease (early H/M: 1.668+/-0.325, late H/M: 1.500+/-0.402) was less than that in multiple system atrophy (early H/M: 2.395+/-0.186, late H/M: 2.530+/-0.391) and controls (early H/M: 2.635+/-0.508, late H/M: 2.575+/-0.635) (early: P<0.0001, late: P<0.0001). There were no significant differences in uptake by the lung, thyroid, or liver in the three groups. Only on early images, uptake in the shoulder in multiple system atrophy (early S/M: 0.473+/-0.78) and Parkinson's disease (early S/M: 0.470+/-0.710) was decreased compared to that in controls (early S/M: 0.560+/-0.118) (P=0.0252). MIBG is reported to be taken up in the terminal part of sympathetic nerves and demonstrates sympathetic nerve activity, especially on late images. The cause of differences between the heart and other parts of the body remains unknown. We consider the following possibilities: (a) differences in the sympathetic nervous system between Parkinson's disease and multiple system atrophy are more subtle in organs other than the heart; (b) the cause of MIBG uptake reduction by the heart in Parkinson's disease involves factors in addition to sympathetic nervous system damage; and (c) MIBG uptake by organs other than the heart involves not only the sympathetic nervous system but also non-neuronal components. In conclusion, MIBG uptake by organs other than the heart cannot differentiate Parkinson's disease from multiple system atrophy at present. PMID- 15893709 TI - Closed-loop analysis of cardiovascular variability in rats under restraint stress. AB - Causal transfer function analysis was applied to the heart rate variability and blood pressure variability in normotensive male Sprague-Dawley rats those were measured before, during, and after acute restraint stress. The causal transfer gain (CTG) from systolic blood pressure (SBP) to RR interval (RRI) and CTG from RRI to SBP were estimated. The mean value of the CTG from SBP to RRI in the low frequency (LF) band (0.27-0.74 Hz) was significantly lower during the restraint period than during the baseline period and remained significantly lower during the recovery period. The mean value of the CTG from RRI to SBP in the LF band, in contrast, was significantly higher during the restraint period than during the baseline period, and during the recovery period it tended to return relatively rapidly to the baseline level. Arterial pressure tended to increase only at the onset of the restraint stress and then to decline not only during the restraint period but also during the recovery period, finally leveling off significantly below the baseline level. These results suggest that restraint stress suppresses the baroreflex control gain not only during the restraint period but also during the recovery period and that this results in the arterial hypotension during the recovery period. PMID- 15893710 TI - Impaired clearance of dying cells in systemic lupus erythematosus. AB - Impaired clearance of apoptotic cell material has been implicated in the pathogenesis of systemic lupus erythematosus (SLE). Besides many other molecules, C1q and DNaseI contribute to an efficient clearance of dying cells. A frequently observed factor in SLE patients is the accumulation of unusually large amounts of apoptotic cells in various tissues. We showed that in a subgroup of patients with SLE, apoptotic cells accumulated in the germinal centers of the lymph nodes. The numbers of tingible body macrophages usually containing engulfed apoptotic nuclei were significantly reduced in these patients. Furthermore, we differentiated macrophages from CD34+ stem cells of SLE patients and NHD in vitro to analyze whether the observed clearance defects are intrinsic. Indeed, macrophages from SLE patients showed a reduced phagocytic capability. Very interestingly, those macrophages from different SLE patients, as well as granulocytes from these patients, showed in part different phagocytic defects, suggesting a heterogeneous clearance defect. We conclude that a failure of clearance in the early phase of apoptosis leads to a secondary necrotic status of the cells. Danger signals are released, modified autoantigens are accessible, and an autoimmune reaction gets started. PMID- 15893711 TI - Clinical and pathological characteristics of Mikulicz's disease (IgG4-related plasmacytic exocrinopathy). AB - Mikulicz's disease (MD) has been considered part of primary Sjogren's syndrome (SS) since Morgan's report in 1953. MD represents a unique condition involving enlargement of the lacrimal and salivary glands, as is also seen in SS; however, MD is characterized by few autoimmune reaction and its good responsiveness to glucocorticoid. Recent reports have shown that the frequency of apoptosis in glands of MD patients is lower when compared with SS. This phenomenon reflects the histologically reversible gland secretion in MD. Elevated IgG4 concentrations in the serum and prominent infiltration by plasmacytes expressing IgG4 in the lacrimal and salivary glands have also been confirmed in MD. Plasma cells expressing IgG4 are also detected in lymph nodes and bone marrow. MD may be a systemic disease, rather than a lacrimal and salivary gland disease. We here propose the new entity "IgG4-related plasmacytic exocrinopathy" and expect future development with regard to its relationship with autoimmune pancreatitis, which similarly presents elevated serum IgG4 levels. PMID- 15893712 TI - Peptidylarginine deiminase type 4, anticitrullinated peptide antibodies, and rheumatoid arthritis. AB - Anticitrullinated peptide antibodies seem to be one of the most clinically reliable serologic markers for rheumatoid arthritis (RA). A genetic approach revealed that one of the citrullinating enzymes has a RA-susceptible variant. Peptidyl citrullination alters the chemical character of peptides and, subsequently, their antigenicity as well. This change in antigenicity of self peptides seems to invoke citrulline-related autoimmunity. Although the precise physiologic role of citrullination is still unknown, accumulating data indicate that citrullination has a definite role in biologic phenomena, along with other posttranslational protein modifications, such as methylation and phosphorylation. In RA synovial tissue, two of five PADI isotypes are known to be expressed, and their expression is regulated at multiple steps: transcription, translation, intracellular localization, and activation/inactivation of PADI proteins. Further investigations on citrulline and PADIs from various aspects will provide a more profound understanding of RA-related autoimmunity. PMID- 15893713 TI - Transient autoimmunity related to maternal autoantibodies: neonatal lupus. AB - Neonatal lupus (NLE) is an autoimmune disease associated with maternal antibodies to Ro/La and characterized by cutaneous lesions, heart block, cardiomyopathy, hepatobiliary disease, and hematologic cytopenias. In most cases, only one organ is affected, although multiple organ involvement is not unusual. Since NLE is presumably caused by maternal autoantibodies, the disease process is transient. However, cardiac NLE, in particular, may be fatal or persistently disabling. Optimal therapy has not yet been determined. Mothers of babies with NLE are often initially asymptomatic, but eventually most develop symptoms of autoimmune disease, particularly diseases associated with anti-Ro/La autoantibodies, such as Sjogren's syndrome and systemic lupus erythematosus. Children who have had NLE are probably at increased risk for autoimmunity later in life, sometimes as early as pre-adolescence, but the magnitude of the risk for the children is not known. Only a small percentage of babies exposed to maternal autoantibodies to Ro and/or La develop NLE. The factors governing which babies develop disease and, if disease develops, which organs will be affected have yet to be fully elucidated. In this review the clinical features, diagnosis, therapy, and prognosis of NLE are discussed, and a summary of experimental data relating to pathogenesis is presented. PMID- 15893714 TI - Xenobiotic-induced autoimmunity and protein aggregation diseases share a common subnuclear pathology. AB - The cell nucleus constitutes a prime target of idiopathic and xenobiotic-induced autoimmunity. Despite of the high prevalence of rheumatic autoimmune diseases, the molecular mechanisms inducing systemic autoimmunity are largely unknown. In appreciation of Rudolf Virchow's cellularpathology, this review introduces the new concept of subnuclear pathology to autoimmune responses against the cell nucleus. Aberrant nucleoplasmic clusters consisting of nuclear autoantigens and proteasomes are observed in xenobiotic-treated cell lines, splenic cells from animal models of xenobiotic-induced autoimmunity, and dendritic cells of scleroderma patients. Aggregation of nuclear proteins in clusters inhibits nuclear functions such as replication and transcription, and induces altered proteasomal degradation of nuclear autoantigens and cellular senescence. Since these modifications of nuclear structure, function and proteolysis resemble the pathology of neurodegenerative protein aggregation diseases, the hypothesis is put forward that xenobiotic-induced autoimmunity may also be a consequence of protein aggregation. PMID- 15893715 TI - Koebner phenomenon in lupus erythematosus with special consideration of clinical findings. AB - The isomorphic response of Koebner can be observed not only in psoriasis, but also in other diseases, such as lichen planus and some systemic diseases including LE (lupus erythematosus) or sarcoidosis. Several clinical findings in LE skin were presented and discussed in this review. The mutually-interactive-, negative-, and internal-Koebner phenomena were introduced and discussed with some speculative views. Many forms of environmental stress on the skin were reported as provocating factors of the Koebner phenomenon, including trauma, scratching, UV-exposure, and various types of dermatitis. Clinical observations of the nature, localization, and movement of lesions should be carefully made. The pathophysiology of the Koebner phenomenon may be classified into two steps. A first non-specific inflammatory step and a second disease-specific step. The inflammatory products released from the first step would be targeted in the second step. In the first step, there could be many substances including cytokines, stress proteins, adhesion molecules, or autoantigens translocated from intra-cellular areas. In the second step after latent periods, there may be disease-specific reactions, including ones by T-cells, B-cells, autoantibodies, and immune deposits, under the restriction of genetic backgrounds. The Koebner phenomenon may prove useful in understanding the pathophysiology of diseases of unknown origin. PMID- 15893716 TI - Immunogenicity of an inflammation-associated product, tyrosine nitrated self proteins. AB - To understand the mechanism leading to autoantibody production, it is of importance to reveal how self-components that are otherwise inactive as antigens acquire immunogenicity. One possible mechanism is the generation of structurally modified self-proteins in apoptotic or inflamed tissues. The post-translational modification of proteins might give rise to the generation of new epitopes to which T and B lymphocytes are not rendered tolerant. Among the protein modifications, this review is focussed on the generation and the immunogenicity of self-proteins carrying 3-nitrotyrosine (NT), an inflammation-associated marker. NT-proteins are generated in vivo by nitration with peroxynitrite, which is formed from nitric oxide and superoxide that are released from activated inflammatory cells. Interestingly, many anti-DNA Abs from autoimmune mice have been shown cross-reactive with NT. Analysis of the immunogenicity of NT-carrying self-proteins has revealed that they elicit both humoral and cellular immune responses in mice. Thus, NT-containing epitopes created on self-proteins may serve as a trigger to impair or bypass immunological tolerance. PMID- 15893717 TI - Improved approach to identify cancer-associated autoantigens. AB - The failure to identify biomarkers of clinical significance for cancer diagnosis and prognosis generated a great deal of skepticism in regard to the usefulness of autoantibody-based methods. SEREX was a major advancement in immunoscreening that resulted in the identification of a large group of autoantigens recognized by cancer sera. However, few SEREX-defined autoantigens have proven to have definitive diagnostic value in clinical practice. Often, the identified antigens are patient-specific rather than tumor-specific and many tumor-associated antigens are rare in expression libraries made from non-autologous cells. Since autoantibodies are part of the normal immune response, it can be difficult to single out tumor-associated antibodies from the scores of irrelevant patient specific responses. In our view, any practical approach for identifying cancer related autoantigens must include an integral strategy for demonstrating tumor relevance early in the screening process. Care must also be taken not to exclude potentially important autoantibodies by pre-screening manipulations to patient sera. We have introduced substantial modifications in SEREX, designed to minimize confounding effects of unrelated autoantibodies and to eliminate steps that preclude the identification of cancer-related autoantigens commonly recognized by cancer sera. In addition, we incorporate methodology to identify candidate antigens that have potential diagnostic or prognostic value prior to their molecular cloning and characterization. PMID- 15893718 TI - The immunological basis of lichenoid tissue reaction. AB - An autoimmune attack by T cells on the epidermis is the primary pathological event in the lichenoid tissue reaction (LTR). The severity of epidermal damage in the LTR is dependent on the relative balance between the intensity and perpetuation of T cell activation and the capacity of epidermal cells to protect from the attack. In natural disease processes, T cells activated by some exogenous agents migrate from the blood to the skin sites and invade the epidermis. Although this epidermotropic migration of T cells resulting in epidermal destruction is a complicated multistep process, this can be bypassed in fixed drug eruption (FDE). Thus, FDE is the most simplified disease model for investigating the pathogenesis of the LTR, in that large numbers of effector CD8+ T cells persist as a stable population in the resting lesions without causing epidermal damage and activation of these T cells resulting in localized epidermal injury can only be induced after administration of the causative drugs. Based on the findings observed with these CD8+ skin-resident T cells, we hypothesize that CD8+ T cells primed during viral infections could evolve into long-lived effector memory phenotype T cells; they would be subsequently trapped either specifically or nonspecifically in the inflamed skin sites and eventually persist as a stable population. Once cross-reacted with exogenous stimuli, such as drug or self antigens, however, they would become effectors of epidermal damage as seen in various lichenoid skin diseases. PMID- 15893719 TI - Socioeconomic factors in lupus erythematosus. AB - For a long time, systemic lupus erythematosus (SLE) was considered a potentially deadly disease. Since the introduction of immunosuppressive therapy, the life expectancy and the quality of life of patients suffering from lupus erythematosus has been dramatically improved. Today, the 5-year survival rate for SLE varies between 50% and 95%. Still, not all patients benefit equally from medical advances. Ethnic and/or socioeconomic minorities show severely disadvantageous prognosis or outcome in various studies. A substantial reduction in the quality of life as well as unemployment are other frequent side effects of this disease. Vocational handicaps related to discoid lupus erythematodes (DLE) was seen in nearly 45% of the patients. Therefore, the management of lupus erythematosus patients requires interdisciplinary cooperation between physicians, psychologists and social workers. The major aim of this article is to summarize the history of lupus erythematosus on the one and the other hand to consider the role of the socioeconomic factors influencing the prognosis of systemic and cutaneous lupus erythematosus. PMID- 15893720 TI - Enhancement of autoantibody pathogenicity by viral infections in mouse models of anemia and thrombocytopenia. AB - Viral infections are involved in the pathogenesis of blood autoimmune diseases such as hemolytic anemia and thrombocytopenia. Although antigenic mimicry has been proposed as a major mechanism by which viruses could trigger the development of such diseases, it is not easy to understand how widely different viruses might induce these blood autoimmune diseases by this sole mechanism. In mice infected with lactate dehydrogenase-elevating virus (LDV), or mouse hepatitis virus, and treated with anti-erythrocyte or anti-platelet monoclonal autoantibodies at a dose insufficient to induce clinical disease by themselves, the infection sharply enhances the pathogenicity of autoantibodies, leading to severe anemia or thrombocytopenia. This effect is observed only with antibodies that induce disease through phagocytosis. Moreover, the phagocytic activity of macrophages from infected mice is increased and the enhancing effect of infection on autoantibody-mediated pathogenicity is strongly suppressed by treatment of mice with clodronate-containing liposomes. Finally, the disease induced by LDV after administration of autoantibodies is largely suppressed in animals deficient for gamma-interferon receptor. Together, these observations suggest that viruses may trigger autoantibody-mediated anemia or thrombocytopenia by activating macrophages through gamma-interferon production, a mechanism that may account for the pathogenic similarities of multiple infectious agents. PMID- 15893721 TI - How cholesterol-dependent cytolysins bite holes into membranes. AB - In a show piece for electron microscopy (EM), Tilley at al. used single-particle cryo-EM to visualize the structural rearrangements in the bacterial toxin pneumolysin that occur when it assembles into a membrane-associated prepore and when the prepore subsequently transitions into a fully membrane-inserted pore. PMID- 15893722 TI - Epigenetic silencing mechanisms in budding yeast and fruit fly: different paths, same destinations. AB - Transcriptional silencing in budding yeast and fruit fly is mediated by fundamentally unrelated proteins that assemble very different chromatin structures. Surprisingly, the repressive mechanisms evolved from these very different materials have similar features, including an epigenetic mode of inheritance and a block to transcription based on interference with the assembly or function of the promoter complex rather than with the binding of gene-specific activators. PMID- 15893723 TI - Eukaryotic transcription activation: right on target. AB - The unambiguous identification of the direct targets of eukaryotic transcriptional activators has been a major challenge in the field. Recently, the authentic targets of several yeast and mammalian activators have been determined, and the results of these studies have important implications for our understanding of transcriptional activation mechanisms. PMID- 15893724 TI - Sequence of steps in ribosome recycling as defined by kinetic analysis. AB - After termination of protein synthesis in bacteria, ribosomes are recycled from posttermination complexes by the combined action of elongation factor G (EF-G), ribosome recycling factor (RRF), and initiation factor 3 (IF3). The functions of the factors and the sequence in which ribosomal subunits, tRNA, and mRNA are released from posttermination complexes are unclear and, in part, controversial. Here, we study the reaction by rapid kinetics monitoring fluorescence. We show that RRF and EF-G with GTP, but not with GDPNP, promote the dissociation of 50S subunits from the posttermination complex without involving translocation or a translocation-like event. IF3 does not affect subunit dissociation but prevents reassociation, thereby masking the dissociating effect of EF-G-RRF under certain experimental conditions. IF3 is required for the subsequent ejection of tRNA and mRNA from the small subunit. The latter step is slower than subunit dissociation and constitutes the rate-limiting step of ribosome recycling. PMID- 15893725 TI - Structural basis for an unexpected mode of SERM-mediated ER antagonism. AB - Tamoxifen is effective for the prevention and treatment of estrogen-dependent breast cancers, but is associated with an increased incidence of endometrial tumors. We report the crystal structure of the estrogen receptor alpha (ERalpha) ligand binding domain (LBD) bound to the structurally similar compound GW5638, which has therapeutic potential and does not stimulate the uterus. Like tamoxifen, GW5638 relocates the carboxy-terminal helix (H12) to the known coactivator-docking site in the ERalpha LBD. However, GW5638 repositions residues in H12 through specific contacts with the N terminus of this helix. In contrast to tamoxifen, the resulting increase in exposed hydrophobic surface of ERalpha LBD correlates with a significant destabilization of ERalpha in MCF-7 cells. Thus, the GW5638-ERalpha LBD structure reveals an unexpected mode of SERM mediated ER antagonism, in which the stability of ERalpha is decreased through an altered position of H12. This dual mechanism of antagonism may explain why GW5638 can inhibit tamoxifen-resistant breast tumors. PMID- 15893726 TI - Identification of FBL2 as a geranylgeranylated cellular protein required for hepatitis C virus RNA replication. AB - We recently reported that Hepatitis C virus (HCV) RNA replication requires one or more geranylgeranylated host proteins. Using a combination of [(3)H]mevalonate labeling, coimmunoprecipitation, and bioinformatic search, we identified a geranylgeranylated host protein required for HCV RNA replication. This protein, FBL2, contains an F box domain and a CAAX motif (CVIL). It forms a stable immunoprecipitable complex with the HCV nonstructural protein 5A (NS5A). The association of FBL2 with NS5A requires the CAAX motif of FBL2, but not the F box. Deletion of the F box created a dominant-negative protein that inhibited replication of HCV RNA when overexpressed in Huh7-K2040 cells; this inhibition was overcome by coexpression of NS5A. siRNA-mediated knockdown of FBL2 mRNA by 70% in Huh7-HP cells reduced HCV RNA by 65%; this reduction was overcome by expression of a cDNA encoding a wobble mutant of FBL2. The current data indicate that geranylgeranylated FBL2 binds to NS5A in a reaction crucial for HCV RNA replication. PMID- 15893727 TI - Nucleophosmin/B23, a nuclear PI(3,4,5)P(3) receptor, mediates the antiapoptotic actions of NGF by inhibiting CAD. AB - Phosphatidylinositol 3,4,5-triphosphate [PI(3,4,5)P(3)] is an essential second messenger implicated in various cellular processes. Cytoplasmic PI(3,4,5)P(3) has been well characterized, but little is known about the physiological role of nuclear PI(3,4,5)P(3). Here, we describe a nuclear PI(3,4,5)P(3) receptor, nucleophosmin (NPM)/B23, that mediates the antiapoptotic effects of NGF by inhibiting DNA fragmentation activity of caspase-activated DNase (CAD). Employing PI(3,4,5)P(3) column and NGF-treated PC12 nuclear extracts, we identified B23 as a nuclear PI(3,4,5)P(3) binding protein. Purification from nuclear extract demonstrates that B23 contributes to DNA fragmentation inhibitory activity. Depletion of B23 from nuclear extracts or knockdown B23 in PC12 cells abolishes NGF-provoked protective effect, whereas overexpression of B23 in PC12 cells prevents apoptosis. Further, hydrolyzing PI(3,4,5)P(3) with PTEN or SHIP abrogates its antiapoptotic activity. Moreover, B23 mutants that can not associate with PI(3,4,5)P(3) fail to prevent DNA fragmentation. Thus, the nuclear B23-PI(3,4,5)P(3) complex regulates the antiapoptotic activity of NGF in the nucleus. PMID- 15893728 TI - The transcriptional coactivator Yes-associated protein drives p73 gene-target specificity in response to DNA Damage. AB - The transcriptional coactivator Yes-associated protein (YAP) has been shown to interact with and to enhance p73-dependent apoptosis in response to DNA damage. Here, we show that YAP requires the promyelocytic leukemia gene (PML) and nuclear body localization to coactivate p73. YAP imparts selectivity to p73 by promoting the activation of a subset of p53 and/or p73 target promoters. Endogenous p73, YAP, and p300 proteins are concomitantly recruited onto the regulatory regions of the apoptotic target gene p53AIP1 only when cells are exposed to apoptotic conditions. Silencing of YAP by specific siRNA impairs p300 recruitment and reduces histone acetylation on the p53AIP1 target gene, resulting in delayed or reduced apoptosis mediated by p73. We also found that YAP contributes to the DNA damage-induced accumulation of p73 and potentiates the p300-mediated acetylation of p73. Altogether, our findings identify YAP as a key determinant of p73 gene targeting in response to DNA damage. PMID- 15893729 TI - Dynamic error correction and regulation of downstream bubble opening by human RNA polymerase II. AB - The nucleotide triphosphate (NTP)-driven translocation hypothesis posits that NTP substrates bind to templated DNA sites prior to translocation into the active site. By using millisecond phase kinetics, we demonstrate this prediction in three different ways. First, we show that, in the presence of the translocation blocker alpha-amanitin, NTPs (but not deoxynucleotide triphosphate [dNTPs]) templated at downstream sites (i + 2 and i + 3) dislodge an active site (i + 1) NTP, which was otherwise fated to complete bond synthesis. Second, we show that NTPs templated at i + 2 and/or i + 3 downstream sites suppress misincorporation errors. Third, we show that NTPs templated at downstream sites stabilize the posttranslocated elongation complex at a stall position. Therefore, at least two NTP substrates pair to DNA templated sites downstream of the active site. These results demonstrate the mechanisms of NTP loading and transcriptional efficiency and fidelity for human RNA polymerase II and indicate regulation of downstream bubble opening by NTPs. PMID- 15893730 TI - Functional characterization of core promoter elements: DPE-specific transcription requires the protein kinase CK2 and the PC4 coactivator. AB - Downstream core promoter elements are an expanding class of regulatory sequences that add considerable diversity to the promoter architecture of RNA polymerase II transcribed genes. We set out to determine the factors necessary for downstream promoter element (DPE)-dependent transcription and find that, against expectations, TFIID and the GTFs are not sufficient. Instead, the protein kinase CK2 and the coactivator PC4 establish DPE-specific transcription in an in vitro transcription system containing TFIID, Mediator, and the GTFs. Chromatin immunoprecipitation analyses using the DPE-dependent IRF-1 and TAF7 promoters demonstrated that CK2, and PC4 are present on these promoters in vivo. In contrast, neither PC4 nor CK2 were detected on the TAF1-dependent cyclin D promoter, which contains a DCE type of downstream element. Our findings also demonstrate that CK2 activity alters TFIID-dependent recognition of DCE sequences. These data establish that CK2 acts as a switch, converting the transcriptional machinery from functioning on one type of downstream element to another. PMID- 15893731 TI - Regulation of tissue-specific and extracellular matrix-related genes by a class I histone deacetylase. AB - Class I histone deacetylases (HDACs) repress transcription by deacetylating histones and have been shown to play crucial roles in mouse, Xenopus, zebrafish, and C. elegans development. To identify the molecular networks regulated by a class I HDAC in a multicellular organism, we carried out a global gene expression profiling study using C. elegans embryos, and identified tissue-specific and extracellular matrix (ECM)-related genes as major HDA-1 targets. Ectopic expression of HDA-1 or C. elegans cystatin, an HDA-1 target identified from the microarray, significantly perturbed mammalian cell invasion. Similarly, RNAi depletion or overexpression of human HDAC-1 also affected cell migration. These findings suggest that HDA-1/HDAC-1 may play a critical, evolutionarily conserved role in regulating the extracellular microenvironment. Because human HDACs are targets for cancer therapy, these findings have significant implications in cancer treatment. PMID- 15893732 TI - Negative control contributes to an extensive program of meiotic splicing in fission yeast. AB - Despite a high frequency of introns in the fission yeast Schizosaccharomyces pombe, regulated splicing is virtually unknown. We present evidence that splicing constitutes a major mechanism for controlling gene expression during meiosis, as 12 of 96 transcripts tested, which encode known components as well as previously uncharacterized ORFs, retain introns until specific times during differentiation. The meiotically spliced pre-mRNAs include two cyclins, rem1 (discovered by Ayte and Nurse) and crs1. Consistent with the use of regulated splicing to block protein production, expression of crs1 in vegetative cells is toxic. Analyses of gene chimeras indicate that splicing is prevented in mitotically growing cells via inhibition, in contrast to the positive control of meiotic splicing in budding yeast. Most strikingly, splicing of crs1 and rem1 is regulated by sequences located outside the coding regions, far from the target introns, a phenomenon previously observed only in metazoans. PMID- 15893733 TI - Human CD69 associates with an N-terminal fragment of calreticulin at the cell surface. AB - CD69 is thought to be a pluripotent signaling molecule expressed on the surface of a number of activated leukocytes including B, T, and NK cells, monocytes, neutrophils, and platelets. While some advances have been made regarding the mechanisms by which CD69 may participate in such diverse functions as cell aggregation, cellular cytotoxicity, and release of cytokines and inflammatory mediators, the most proximal links of signal initiation have not been identified. Our study has identified, by immunoprecipitation and direct protein sequencing (LC/MS/MS), binding of CD69 to an N-terminal protein fragment of calreticulin expressed on the cell surface of human PBMCs. Given the recently identified roles calreticulin plays in cell adhesion and angiogensis, the identification of CD69 binding directly to calreticulin may provide insights into mechanism(s) by which CD69 or other CD69 family members, i.e., LLT1 and AICL participates in such diverse functions. PMID- 15893734 TI - Oestrogen receptor-immunoreactive neurons in the trigeminal sensory system of male and cycling female rats. AB - Many common craniofacial pain conditions are more prevalent in women than men and may be related to the phase of the menstrual cycle. Long-term effects of oestrogen in the nervous system are produced by receptor-mediated [oestrogen receptor alpha (ERalpha) and beta (ERbeta) isoforms] mechanisms; however, it is not known if the distribution of ER-positive neurons in the trigeminal system is similar in males and females. Quantitative immunocytochemistry was used to compare the distribution of ERalpha-labelled neurons in the trigeminal brainstem complex (TBC) and ganglion of male and female rats at different stages of the oestrous cycle. A high density of ERalpha-labelled neurons was seen in the superficial laminae (I-III) throughout the trigeminal subnucleus caudalis (Vc) and the upper cervical dorsal horn. Counts of ERalpha-positive neurons in laminae I-III were similar for prooestrous and dioestrous females, while males had fewer cells. The deeper laminae (IV-V) of the Vc and the cervical dorsal horn had few ERalpha-positive neurons in all groups. At the region surrounding the central canal at caudal levels of the Vc, prooestrous females had more ERalpha-positive neurons than dioestrous females or males. Few labelled cells were seen rostral to the trigeminal subnucleus interpolaris/caudalis transition region (Vi/Vc) in any group. In the trigeminal ganglion, prooestrous and dioestrous females had a moderate (8-10%) number of nuclear-labelled small or medium-sized neurons, while males had fewer labelled cells (4.5%). Qualitatively, the pattern of staining for ERbeta was similar, although weaker, than for ERalpha in the trigeminal dorsal horn or ganglion. These results were consistent with the hypothesis that oestrogen acts through trigeminal ganglion cells and caudal portions of the Vc to modulate sensory and autonomic aspects of craniofacial pain in a sex-related manner. PMID- 15893735 TI - The effect of age on morphology and eruption of the lower incisors in mature rats. AB - OBJECTIVE: This study investigated the effect of age on tooth and periodontium morphology and on tooth eruption in the lower incisors of 28 female rats, initial age 10 weeks. DESIGN: Morphometric data of the incisors in three young mature rats (weight 199+/-2.4 g) were compared to that of three 18-week-old (weight 260+/-5 g) animals. The rate of eruption was monitored during 12 months in 11 rats with all teeth in occlusion and in 11 rats with one lower incisor kept constantly out of occlusion. RESULTS: : The incisors continued to enlarge after the animals reached maturity with the size of the teeth increasing both longitudinally and circumferentially. Tooth volume increased by 21%, while that of the periodontal ligament was enlarged by only 12%. The rate of impeded eruption declined steadily from a mean value of 542+/-49 microm/day during the first experimental month to a mean value of 443+/-25 microm/day during the concluding month (p<0.05). The eruption rate of the teeth exceeded that of attrition rate by 2.4%. The unimpeded eruption proceeded steadily throughout the experimental year, at a mean rate of 811+/-12. CONCLUSIONS: Apparently in mature rats age has no effect on the genetic potential as expressed by the unimpeded eruption. In the impeded teeth age does affect the phenotypic expression of the eruptive function governed by occlusal forces as well as by the form and size ratio of the tooth and its PDL. PMID- 15893736 TI - Plasmid vectors harboring cellular promoters can induce prolonged gene expression in hematopoietic and mesenchymal progenitor cells. AB - Although prolonged transgene expression in progenitor cells might be desirable for modified cell therapy, the viral promoter-based expression vector tends to promote transgene expression only for a limited period. Here, we examined the ability of cellular promoters from elongation factor-1alpha (EF-1alpha) and ubiquitin C to drive gene expression in hematopoietic TF-1 and mesenchymal progenitor cells. We compared the expression levels and duration of a model gene, interleukin-2, generated by the cellular promoters to those by the cytomegalovirus (CMV) promoter. The EF-1alpha and ubiquitin C promoters drove prolonged gene expression in hematopoietic TF-1 and mesenchymal progenitor cells, whereas the CMV promoter did not. At day 7 after transfection in TF-1 cells, the mRNA expression levels of interleukin-2 driven by the EF-1alpha and ubiquitin C promoters were 118- and 56-fold higher, respectively, than those driven by the CMV promoter. Similarly, in mesenchymal progenitor cells, the expression levels of interleukin-2 driven by the EF-1alpha and ubiquitin C promoters were 98- and 20-fold higher, respectively, than that driven by the CMV promoter-encoding plasmid. Moreover, the ubiquitin C promoter directed higher levels of green fluorescence protein expression in mesenchymal progenitor cells than did the CMV promoter. These results indicate that the use of cellular promoters such as those for EF-1alpha and ubiquitin C might direct prolonged gene expression in hematopoietic and mesenchymal progenitor cells. PMID- 15893737 TI - Inhibitors of cation-chloride-cotransporters affect hypoxic/hypoglycemic injury in hippocampal slices. AB - Electroneutral cation-chloride cotransporters are abundantly expressed in the brain and are involved in the regulation of the intracellular Cl(-) concentration and thus gamma-aminobutyric acid-dependent inhibition of neuronal excitability. As yet there is little evidence whether or not Na(+)-K(+)-2Cl(-) or K(+)-Cl(-) cotransporters are involved in neuronal hyperexcitability and death in cerebral ischemia. In this study, by measuring propidium iodide staining in organotypic hippocampal slice cultures from young rats and population spike recovery in acutely isolated hippocampal slices from adult rats after a hypoxic/hypoglycemic insult, we were able to assess if cation-chloride cotransport inhibitors reduce neuronal injury. The Na(+)-K(+)-2Cl(-) cotransport inhibitor bumetanide in the range of 1-10 microM reduced neuronal damage in the slice cultures by 25%, but did not affect population spike recovery in acutely isolated slices. In contrast the K(+)-Cl(-) cotransport inhibitor [(dihydroindenyl)oxy] alkanoic acid (DIOA, 100 microM) significantly diminished the restitution of the population spikes from 33% before to 8% after hypoxia/hypoglycemia and increased the damage in the slice cultures by 60%. Consequently, our data suggest that the Na(+)-K(+)-2Cl(-) cotransporter may contribute to neuronal injury and that the activity of the K(+) Cl(-) cotransporters is an intrinsic protective mechanism of neurons against ischemic damage. PMID- 15893738 TI - Protective effect of donepezil against Abeta(1-40) neurotoxicity in rat septal neurons. AB - Donepezil, a potent acetylcholinesterase (AChE) inhibitor used for the treatment of Alzheimer's disease (AD), is thought to have a neuroprotective effect in AD patients. Because a deficit in cholinergic neurotransmission is a major feature in AD, and amyloid-beta (Abeta) accumulation has been proposed as a possible causative phenomenon, we were interested to examine the effect of donepezil on Abeta(1-40) induced neurotoxicity in primary cultures of rat septal neurons. Using immunohistochemical staining, almost all the neurons were found to be positive for vesicular acetylcholine transporter (VAChT) in these septal cultures. Septal neuronal cells were cultured for 7 days and then 15 micromol/L of Abeta(1-40) was added to the cell medium for 48 h. The cultured septal neurons were highly susceptible to Abeta toxicity, as shown by morphological examination and lactate dehydrogenase (LDH) assay. Donepezil concentration-dependently reduced the LDH efflux induced by Abeta(1-40), and the effect was significant at 100 nmol/L and above. Donepezil decreased both the negative peak at around 215 nm in the circular dichroism (CD) spectrum and the fluorescence intensity of thioflavin T in the presence of Abeta(1-40). These results suggest that donepezil exerts a neuroprotective effect by reducing the amount of the toxic form of Abeta fibrils in septal neuron cultures. These findings support the idea that the clinical efficacy of donepezil in AD is due to not only activation of cholinergic transmission, but also attenuation of neuronal damage. PMID- 15893739 TI - Regulation of myeloid leukemia factor-1 interacting protein (MLF1IP) expression in glioblastoma. AB - The myelodysplasia/myeloid leukemia factor 1-interacting protein MLF1IP is a novel gene which encodes for a putative transcriptional repressor. It is localized to human chromosome 4q35.1 and is expressed in both the nuclei and cytoplasm of cells. Northern and Western blot analyses have revealed MLF1IP to be present at very low amounts in normal brain tissues, whereas a number of human and rat glioblastoma (GBM) cell lines demonstrated a high level expression of the MLF1IP protein. Immunohistochemical analysis of rat F98 and C6 GBM tumor models showed that MLF1IP was highly expressed in the tumor core where it was co localized with MLF1 and nestin. Moreover, MLF1IP expression was elevated in the contralateral brain where no tumor cells were detected. These observations, together with previous data demonstrating a role for MLF1IP in erythroleukemias, suggest a possible function for this protein in glioma pathogenesis and potentially in other types of malignancies. PMID- 15893740 TI - Activation-flow coupling during graded cerebral ischemia. AB - Most functional neuroimaging techniques rely on activation-flow coupling (AFC) to detect changes in regional brain function, but AFC responses may also be altered during pathophysiological conditions such as ischemia. To define the relationship between progressive ischemia and the AFC response, graded levels of cerebral blood flow reduction were produced using a rat compression ischemia model, and the cerebral hemodynamic response to forepaw stimulation was measured. Graded levels of cortical ischemia of the somatosensory cortex were induced in male Sprague-Dawley rats (n = 16) by compressing the intact dura with a 4-mm-diameter cylinder equipped with a laser-Doppler probe, combined with ipsilateral common carotid artery occlusion. At each level of CBF reduction, electric forepaw stimulation was conducted, and signal-averaged laser Doppler and evoked potential responses were recorded. A visible AFC response was present at all levels of CBF reduction (0-90% reduction from baseline), and the temporal characteristics of the response appeared largely preserved. However, the amplitude of the AFC response began to decline at levels of mild ischemia (10% flow reduction) and progressively decreased with further CBF reduction. The amplitude of the evoked response appeared to decrease in concert with the AFC amplitude and appeared to be equally sensitive to ischemia. AFC appears to be a sensitive marker for cerebral ischemia, and alterations in the AFC response occur at CBF reductions above the accepted thresholds for infarction. However, the AFC response is also preserved when flow is reduced below ischemic thresholds. PMID- 15893741 TI - Clinical proteomics: translation of new insights into clinical practice. Proceedings of the International Conference on Laboratory Medicine. October 29, 2004. Padua, Italy. PMID- 15893742 TI - Postprandial hypertriglyceridemia and oxidative stress in patients of type 2 diabetes mellitus with macrovascular complications. AB - BACKGROUND: Oxidative stress has been implicated in vascular complications of diabetes mellitus (DM). This study aims to evaluate the relationship between postprandial hypertriglyceridemia (PP-HTG) and oxidative stress in Indian patients of type 2 DM with macrovascular complications. METHODS: Plasma triglycerides (TG), thiobarbituric acid reactive substances (TBARS), erythrocyte reduced glutathione (GSH) and superoxide dismutase (SOD) were measured in fasting and postprandial (PP) state at 2, 4, 6 and 8 h after a high fat meal challenge in controls (Group I) and patients of type 2 DM without (Group II) and with macrovascular complications (Group III). RESULTS: Postprandial TGs increased significantly in patients with type 2 DM, which showed an exaggerated response to high fat meal challenge in Group III as compared to Group II. Highest PP-TBARS were also observed in Group III which correlated positively with TG. However, GSH and SOD were lower in both groups of diabetics as compared to controls. CONCLUSIONS: The magnitude of PP-HTG appears to be the major determinant of oxidative stress in type 2 DM, which along with a compromised antioxidant status may lead to endothelial dysfunction and macrovascular complications. PMID- 15893743 TI - Evaluation and analysis of exposure levels of di(2-ethylhexyl) phthalate from blood bags. AB - BACKGROUND: The US FDA and The Ministry of Health, Labor and Welfare of Japan have indicated that the risk assessment of di(2-ethylhexyl) phthalate (DEHP) released from polyvinyl chloride (PVC) medical devices requires immediate attention. In particular, the analysis of the exposure to DEHP from blood bags is very important for medical treatment. However, human exposure to DEHP via blood transfusion remains poorly understood. We evaluated DEHP and mono(2-ethylhexyl) phthalate (MEHP) levels, migration patterns, and metabolism in blood products for the detailed assessment of exposure to DEHP. METHODS: A method that is based on column-switching liquid chromatography-electrospray mass spectrometry (LC-MS) coupled with on-line extraction was used for the direct analysis of DEHP and MEHP in the blood products. From the Japanese Red Cross Society, 78 blood products (red blood cell concentrate: n=18, irradiated red blood cell concentrate: n=18, whole blood: n=18, blood platelet: n=18, and frozen plasma: n=6) were sampled in January 2003 for use in this study. RESULTS: The detection levels of DEHP and MEHP ranged from 1.8 to 83.2 microg/ml and from 0.1 to 9.7 microg/ml, respectively. The levels of MEHP and DEHP in the blood products were increased with increasing storage time. In addition, whole blood products in PVC bags had the highest DEHP levels compared to the other blood products. Our results indicate that the maximum level of human exposure to DEHP released from blood bags is 0.7 mg/kg weight/time. CONCLUSION: This first quantitative evidence may be useful for the risk assessment of DEHP released from blood bags. PMID- 15893744 TI - Expression analysis and prognostic significance of human kallikrein 11 in prostate cancer. AB - BACKGROUND: Kallikrein 11 (KLK11) is a newly discovered human kallikrein gene that is mainly expressed in the central nervous system and endocrine tissues. KLK11 has two alternative splicing isoforms, known as the brain type and prostate type. Many members of the human kallikrein gene family are differentially expressed in cancer and a few have potential as diagnostic/prognostic markers. METHODS: In the present study, the expression of prostate type variant of KLK11 gene was analyzed by RT-PCR in 66 prostate cancer tissues. Tumors were pulverized, total RNA was extracted, and cDNA was prepared by reverse transcription. KLK11 was amplified by PCR using gene specific primers and its identity was verified by sequencing. Prostate tissues were then classified as KLK5 positive or negative based on eithidium bromide staining in agarose gels and image analysis. RESULTS: KLK11 was found to be highly expressed in 43/66 (65%) of prostate cancer samples. We found a significant negative relationship between KLK11 expression and Gleason score (p = 0.004) and disease stage (p = 0.038). Serum total PSA concentration was found to be lower in patients with overexpression of KLK11 (p = 0.044). CONCLUSIONS: We conclude that down regulation of the KLK11 gene in advanced and more aggressive tumors may open the possibility of being used as a future biological marker distinguishing the tumor aggressiveness as well as a useful prognostic biomarker for prostate cancer. PMID- 15893745 TI - Impact of missing data in evaluating artificial neural networks trained on complete data. AB - This study investigated the impact of missing data in the evaluation of artificial neural network (ANN) models trained on complete data for the task of predicting whether breast lesions are benign or malignant from their mammographic Breast Imaging and Reporting Data System (BI-RADS) descriptors. A feed-forward, back-propagation ANN was tested with three methods for estimating the missing values. Similar results were achieved with a constraint satisfaction ANN, which can accommodate missing values without a separate estimation step. This empirical study highlights the need for additional research on developing robust clinical decision support systems for realistic environments in which key information may be unknown or inaccessible. PMID- 15893746 TI - Up-regulation of GABA(B) receptors by chronic administration of the GABA(B) receptor antagonist SCH 50,911. AB - Chronic treatment of mice with the specific gamma-aminobutyric acid(B) (GABA(B)) receptor antagonist (2S)(+)-5,5-dimethyl-2-morpholineacetic acid (SCH 50,911) increased both the number of GABA(B) receptors in the whole brain (measured as [3H]CGP 54626 [S-(R,R)]-3-[[1-(3,4-dichlorophenyl)amino]-2 hydroxypropyl](cyclohexylmethyl)phosphinic acid hydrochloride binding) and the ability of baclofen to activate GABA(B) receptor coupled G-protein (measured as % reduction of the EC50 of baclofen to activate [35S]GTP(gamma)S binding). The results indicate that persistent blockade of GABA(B) receptors leads to their compensatory up-regulation and suggest that GABA(B) receptors are tonically activated by endogenous GABA. PMID- 15893747 TI - Nicotine produces antidepressant-like actions: Behavioral and neurochemical evidence. AB - Converging lines of evidence indicate the involvement of nicotinic acetylcholine receptors in depressive illness and antidepressant drug action. We investigated the effects of sub-chronic and chronic treatment with imipramine, nicotine and their combination on: (a) the ability of a dopamine-mimetic challenge to produce locomotor stimulation and (b) cortical density of beta-adrenoceptors. One week of treatment with imipramine (10 mg/kg, twice daily) did not result in an altered response to the apomorphine (0.15 mg/kg) challenge, but after 2 weeks, the imipramine-treated rats demonstrated hyperactivity. Conversely, such increased locomotor response was observed in rats treated with nicotine (0.4 mg/kg, twice daily) for 1 but not for 2 weeks. Groups treated with nicotine+imipramine for 1 and 2 weeks demonstrated equally high hyperactivity in response to the apomorphine challenge. This effect was not different from the effects of 1-week treatment with nicotine or 2-week treatment with imipramine. The density of beta adrenoceptors was equally decreased by 2 (but not 1) weeks of the treatment with imipramine, nicotine and their combination. The present behavioral and neurochemical data suggest the antidepressant-like effect of the chronic treatment with nicotine. It appears that the potentiation of the dopamine-mimetic induced hyperactivity cannot be explained by beta-adrenoceptor down-regulation. PMID- 15893748 TI - Etheric biology. AB - Several observations made during the course of studies on stealth-adapted viruses are explainable by a pervasive, energy-rich, ether environment. Activation of an alternative cellular energy (ACE) pathway provides stealth virus damaged cells with a repair mechanism that is independent of the cellular immune response. ACE activation can also assist in the systemic healing of infections caused by conventional viruses such as Herpes simplex virus, Herpes zoster virus and human papillomavirus. ACE pigments convert conventional forms of physical energies into a biological cell healing energy, the nature of which is still uncertain. More recent studies suggest that ACE pigments may also capture etheric energy. In addition to cellular repair, ACE pigment activation can lead to the biogenesis of lipid-like chemical structures. ACE pigment and virus culture healing activities were also seen with several natural products, including a homeopathic formulation. A colloidal silver solution appeared to facilitate the transmission of ACE and to enhance its biosynthetic activity. These results open a window into a greater understanding of a fundamental force of nature of potential therapeutic importance. PMID- 15893749 TI - Muscle costameric protein, Chisel/Smpx, associates with focal adhesion complexes and modulates cell spreading in vitro via a Rac1/p38 pathway. AB - The murine X-linked gene Chisel (Csl/Smpx) encodes a 9-kDa protein that associates in heart and skeletal muscle cells with the costameric cytoskeleton, implicated in maintaining muscle integrity and responses to biomechanical stress. After expression in C2C12 myoblasts, MYC epitope-tagged Csl co-localized with actin networks at peripheral membranes, and with focal adhesion proteins vinculin, paxillin, integrin beta1, and the small GTPase Rac1. Csl could be co immunoprecipitated with vinculin from extracts of C2C12 cells and native muscle. MYC-Csl induced cell spreading and lamellipodia formation in C2C12 cells at the expense of filopodia, suggestive of modulation of Rac1 activity. Lamellipodia formation was indeed Rac1-dependent, and in MYC-Csl cells replated on fibronectin, Rac1 activity was increased relative to controls. Expression of MYC Csl led to an increased association between vinculin and p34, a subunit of the Arp2/3 actin nucleation complex, a Rac1-dependent event. Induced cell spreading was also dependent upon p38 kinases that act downstream of Rac1 to control the actin capping activity of heat shock protein 27. Our data suggest that Csl localizes to the costameric cytoskeleton of muscle cells through an association with focal adhesion proteins, where it may participate in regulation of cytoskeletal dynamics through the Rac1-p38 pathway. PMID- 15893750 TI - MZF-1 and DbpA interact with DNase I hypersensitive sites that correlate with expression of the human MUC1 mucin gene. AB - The MUC1 mucin is a large membrane-tethered glycoprotein that shows differential expression in many adenocarcinomas, where it contributes to their invasive and metastatic properties. We previously identified DNase I hypersensitive sites at 750 and -250 bp in the human MUC1 gene promoter and showed concordance between the -250 site and MUC1 mRNA levels in vivo. Transient expression assays using promoter constructs, in which the core DHS was deleted, to drive reporter gene expression revealed in vivo evidence for their activity. DNase I footprinting using nuclear extracts from HPAF human pancreatic carcinoma cells and MCF7 breast carcinoma cells identified three protein-binding elements in these regions ( 250FP1, FP2 and -750FP). Electrophoretic mobility shift assays detected several complexes between HPAF nuclear proteins and labeled FP DNA probes. Southwestern blots and UV cross-linking experiments identified myeloid zinc finger-1 (MZF-1) as a candidate transcription factor among proteins binding to the -250FP1 and FP2 sequences. Another candidate that was identified by screening an HPAF cDNA expression library with the -250FP1 probe is DNA binding protein A (DbpA). Exogenous DbpA expression in COS-7 cells was accompanied by upregulation of MUC1 promoter activity via the -250 DHS, suggesting that DbpA binding to the -250 DHS can influence human MUC1 gene expression. PMID- 15893751 TI - p85 beta-PIX is required for cell motility through phosphorylations of focal adhesion kinase and p38 MAP kinase. AB - Lysophosphatidic acid (LPA) mediates diverse biological responses, including cell migration, through the activation of G-protein-coupled receptors. Recently, we have shown that LPA stimulates p21-activated kinase (PAK) that is critical for focal adhesion kinase (FAK) phosphorylation and cell motility. Here, we provide the direct evidence that p85 beta-PIX is required for cell motility of NIH-3T3 cells by LPA through FAK and p38 MAP kinase phosphorylations. LPA induced p85 beta-PIX binding to FAK in NIH-3T3 cells that was inhibited by pretreatment of the cells with phosphoinositide 3-kinase inhibitor, LY294002. Furthermore, the similar inhibition of the complex formation was also observed, when the cells were transfected with either p85 beta-PIX mutant that cannot bind GIT or dominant negative mutants of Rac1 (N17Rac1) and PAK (PAK-PID). Transfection of the cells with specific p85 beta-PIX siRNA led to drastic inhibition of LPA-induced FAK phosphorylation, peripheral redistribution of p85 beta-PIX with FAK and GIT1, and cell motility. p85 beta-PIX was also required for p38 MAP kinase phosphorylation induced by LPA. Finally, dominant negative mutant of Rho (N19Rho)-transfected cells did not affect PAK activation, while the cells stably transfected with p85 beta-PIX siRNA or N17Rac1 showed the reduction of LPA-induced PAK activation. Taken together, the present data suggest that p85 beta-PIX, located downstream of Rac1, is a key regulator for the activations of FAK or p38 MAP kinase and plays a pivotal role in focal complex formation and cell motility induced by LPA. PMID- 15893752 TI - Regulation of neuronal and glial galectin-1 expression by peripheral and central axotomy of rat primary afferent neurons. AB - Galectin-1 (Gal1) is an endogenously-expressed protein important for the embryonic development of the full complement of primary sensory neurons and their synaptic connections in the spinal cord. Gal1 also promotes axonal regeneration following peripheral nerve injury, but the regulation of Gal1 by axotomy in primary afferent neurons has not yet been examined. Here, we show by immunohistochemistry and in situ hybridization that Gal1 expression is differentially regulated by peripheral nerve injury and by dorsal rhizotomy. Following peripheral nerve injury, the proportion of Gal1-positive DRG neurons was increased. An increase in the proportion of large-diameter DRG neurons immunopositive for Gal1 was paralleled by an increase in the depth of immunoreactivity in the dorsal horn, where Gal1-positive terminals are normally restricted to laminae I and II. Dorsal rhizotomy did not affect the proportions of neurons containing Gal1 mRNA or protein, but did deplete the ipsilateral dorsal horn of Gal1 immunoreactivity, indicating that it is transported centrally by dorsal root axons. Dorsal rhizotomy also resulted in an increase in Gal1 mRNA the nerve peripheral to the PNS-CNS interface (likely within Schwann cells and/or macrophages), and to a lesser extent within deafferented spinal cord regions undergoing Wallerian degeneration. This latter increase was notable in the dorsal columns and along the prior trajectories of myelinated afferents into the deeper dorsal horn. These results show that neuronal and glial expressions of Gal1 are tightly correlated with regenerative success. Thus, the differential expression pattern of Gal1 following peripheral axotomy and dorsal rhizotomy suggests that endogenous Gal1 may be a factor important to the regenerative response of injured axons. PMID- 15893753 TI - Trypanosoma brucei s.l.: characterisation of stocks from Central Africa by PCR analysis of mobile genetic elements. AB - To better understand the epidemiology of sleeping sickness in the Central African sub-region, notably the heterogeneity of Human African Trypanosomiasis (HAT) foci, the mobile genetic element PCR (MGE-PCR) technique was used to genotype Trypanosoma brucei s.l. (T. brucei s.l.) isolates from this sub-region. Using a single primer REV B, which detects positional variation of the mobile genetic element RIME, via amplification of flanking regions, MGE-PCR revealed a micro genetic variability between Trypanosoma brucei gambiense (T. b. gambiense) isolates from Central Africa. The technique also revealed the presence of several T. b. gambiense genotypes and allowed the identification of minor and major ubiquitous genotypes in HAT foci. The presence of several T. b. gambiense genotypes in HAT foci may explain the persistence and the resurgence phenomena of the disease and also the epidemic and the endemic status of some Central African sleeping sickness foci. The MGE-PCR technique represents a simple, rapid, and specific method to differentiate Central African T. brucei s.l. isolates. PMID- 15893754 TI - Identification of B cell adaptor for PI3-kinase (BCAP) as an Abl interactor 1 regulated substrate of Abl kinases. AB - In previous work we showed that Abl interactor 1 (Abi-1), by linking enzyme and substrate, promotes the phosphorylation of Mammalian Enabled (Mena) by c-Abl. To determine whether this mechanism extends to other c-Abl substrates, we used the yeast two-hybrid system to search for proteins that interact with Abi-1. By screening a human leukocyte cDNA library, we identified BCAP (B-cell adaptor for phosphoinositide 3-kinase) as another Abi-1-interacting protein. Binding experiments revealed that the SH3 domain of Abi-1 and the C-terminal polyproline structure of BCAP are involved in interactions between the two. In cultured cells, Abi-1 promoted phosphorylation of BCAP not only by c-Abl but also by v Abl. The phosphorylation sites of BCAP by c-Abl were mapped to five tyrosine residues in the C-terminal region that are well conserved in mammals. These results show that Abi-1 promotes Abl-mediated BCAP phosphorylation and suggest that Abi-1 in general coordinates kinase-substrate interactions. PMID- 15893755 TI - Analysis of telomeric single-strand overhang length in human endometrial cancers. AB - The 3' single-strand telomeric overhang (3'-OH) is a key component of telomere structure. Although telomere length has been well analyzed in a variety of human cancers, no information is available on the 3'-OH length in cancers. In the present study, we examined the 3'-OH length in normal and malignant endometria using telomere-oligonucleotide ligation assay. Although 3'-OH lengths varied among patients, 3'-OH length observed in endometrial cancers was significantly shorter than that found in samples derived from normal endometria (P < 0.001: Student's t-test), suggesting that erosion of 3'-OH length induces impaired telomeric integrity and genomic instability, leading to carcinogenesis. Interestingly, we found that the most aggressive subtypes of endometrial cancers harbored significantly longer 3'-OH length than those with non-aggressive subtypes (P < 0.001: Sheffe's test), suggesting that cancer cells with long 3'-OH length have growth advantage due to their stabilized telomere ends. In contrast, we failed to observe an association between overall telomere length and any clinicopathological characteristics of endometrial cancers. These findings suggest that erosion of 3'-OH length, rather than overall telomere length, play roles in endometrial carcinogenesis. Furthermore, long 3'-OH may serve as a molecular marker for aggressive phenotype of tumors. PMID- 15893756 TI - Analysis of dyslipidemia in patients on chronic hemodialysis in Catalonia. AB - Chronic hemodialysis patients show a high incidence and prevalence of cardiovascular disease of multifactorial etiology and an association between dyslipidemia and accelerated atherosclerosis. We analyzed characteristics of dyslipidemia in 1824 hemodialysis patients (59% men; mean age 65 +/- 15 years) in Catalonia and identified risk factors by logistic regression. Prevalence of dyslipidemia was high (63%). Most frequent lipid alterations were decreased HDL cholesterol (40%), hypertriglyceridemia (31%) and hypercholesterolemia (19%). Total cholesterol/HDL ratio was elevated in 23%. Body mass index (OR 1.08; 95% CI 1.05-1.11), diabetes mellitus (1.4; 1.09-1.79), ischemic heart disease (1.38, 1.08-1.75) and stroke (1.30; 1.0-1.69) were independent factors associated with dyslipidemia. Lengthy time (> 7 years) on dialysis (0.77; 0.59-0.99) and female sex (0.78; 0.64-0.96) were independent protective factors. A significant reduction in the risk of developing dyslipidemia was observed after the age of 50. Lipid-lowering drug use was low (19%), with statins being the most frequent (83%). The percentage of patients reaching target LDL levels according to individual cardiovascular risk (ATPIII) was unsatisfactory, particularly in high risk patients (52%). In light of the high prevalence of dyslipidemia and low adherence to target LDL goals, we conclude that strict control of dyslipidemia should be included in cardiovascular risk prevention strategies for chronic hemodialysis patients. PMID- 15893757 TI - Antioxidants and endothelium protection. AB - The endothelium is a complex organ system that controls the homeostasis of the vasculature by integrating signals between the vascular wall and the vessel lumen. Under physiological conditions, it maintains a normal vascular tone and blood fluidity by elaborating a variety of factors, such as nitric oxide, prostacyclin and endothelin. However, in pathological situations the endothelium can also modify its phenotype facilitating vasoconstriction, inflammation, and thrombotic events. These abnormal responses manifest in different clinical settings, such as hypercholesterolemia, hypertension, diabetes mellitus, and occur in the absence of any morphological change of the vessel. The etiology of these altered endothelial functions is multi-factorial, and the mechanisms underlying them are complex and not yet fully elucidated. Today, there is substantial evidence that many endothelial functions are sensitive to the presence of reactive oxygen species and subsequent oxidative stress. Here, I will review the increasing number of studies showing that exogenous antioxidants can modulate the endothelium-dependent vasodilation responses, the homeostatic endothelium-leukocyte interactions, the balance between pro- and anti-thrombotic properties, and the vascular apoptotic responses. Finally, the non-antioxidant activities of some antioxidant will also be described. PMID- 15893758 TI - Lightweight low-profile nine-accelerometer package to obtain head angular accelerations in short-duration impacts. AB - Despite recognizing the importance of angular acceleration in brain injury, computations using data from experimental studies with biological models such as human cadavers have met with varying degrees of success. In this study, a lightweight and a low-profile version of the nine-accelerometer system was developed for applications in head injury evaluations and impact biomechanics tests. The triangular pyramidal nine-accelerometer package (PNAP) is precision machined out of standard aluminum, is lightweight (65 g), and has a low profile (82 mm base width, 35 mm vertex height). The PNAP assures accurate orthogonal characteristics because all nine accelerometers are pre-aligned and attached before mounting on a human cadaver preparation. The feasibility of using the PNAP in human cadaver head studies is demonstrated by subjecting a specimen to an impact velocity of 8.1 m/s and the resultant angular acceleration peaked at 17 krad/s2. The accuracy and the high fidelity of the PNAP device at high and low angular acceleration levels were demonstrated by comparing the PNAP-derived angular acceleration data with separate tests using the internal nine accelerometer head of the Hybrid III anthropomorphic test device. Mounting of the PNAP on a biological specimen such as a human cadaver head should yield very accurate angular acceleration data. PMID- 15893759 TI - Development of a sensitive, colorometric microarray assay for allergen-responsive human IgE. AB - With the existence of several thousand unique human allergens, a multiplex format, such as protein microarrays, is an attractive option for allergy screening. To determine the feasibility and sensitivity of using an enzyme-based, colorimetric protein microarray assay, three common allergens (mold, dust-mite, grass) were arrayed and added sera assayed for responsive human IgE. Normal, low positive, and negative control samples were assayed to determine optimal reaction parameters. Sensitivity of the assay (in international units, IU) was determined by constructing a standard curve using World Health Organization (WHO) standards. The system described here can reliably detect allergen-specific IgE below 0.35 IU, the current WHO standard cutoff. By taking advantage of the sensitivity of enzyme-linked immunosorbent assays (ELISAs) and the multiplex format of microarrays, we have achieved a high throughput system, capable of screening patients for allergen-susceptibility with optimal sensitivity. PMID- 15893760 TI - Horizontal and vertical transmission of wild-type and recombinant Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus. AB - Transmission plays a central role in the ecology of baculoviruses and the population dynamics of their hosts. Here, we report on the horizontal and vertical transmission dynamics of wild-type Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV-WT) and a genetically modified variant (HaSNPV-AaIT) with enhanced speed of action through the expression of an insect selective scorpion toxin (AaIT). In caged field plots, horizontal transmission of both HaSNPV variants was greatest when inoculated 3rd instar larvae were used as infectors, transmission was intermediate with 2nd instar infectors and lowest with 1st instar infectors. Transmission was greater at a higher density of infectors (1 per plant) than at a lower density (1 per 4 plants); however, the transmission coefficient (number of new infections per initial infector) was lower at the higher density of infectors than at the lower density. HaSNPV-AaIT exhibited a significantly lower rate of transmission than HaSNPV-WT in the field cages. This was also the case in open field experiments. In the laboratory, the vertical transmission of HaSNPV-AaIT from infected females to offspring of 16.7+/ 2.1% was significantly lower than that of HaSNPV-WT (30.9+/-2.9%). Likewise, in the field, vertical transmission of HaSNPV-AaIT (8.4+/-1.1%) was significantly lower than that of HaSNPV-WT (12.6+/-2.0%). The results indicate that the recombinant virus will be transmitted at lower rates in H. armigera populations than the wild-type virus. This may potentially affect negatively its long-term efficacy as compared to wild-type virus, but contributing positively to its biosafety. PMID- 15893761 TI - Seasonal dynamics of entomopathogenic nematodes of the genera Steinernema and Heterorhabditis as a response to abiotic factors and abundance of insect hosts. AB - The seasonal dynamics of entomopathogenic nematodes (EPNs) of the genus Steinernema and Heterorhabditis were studied during one season in meadow and oak wood habitats, in the vicinity of Ceske Budejovice, Czech Republic. The influences of soil temperature, moisture, and abundance of suitable hosts on EPN dynamics were investigated. The host range of these nematodes, in both habitats was also observed. A total of four EPN species were found in both habitats. Steinernema affine was the dominant species both in oak wood and in meadow. Additionally, the oak wood habitat was inhabited by S. kraussei and S. weiseri; the meadow habitat by Heterorhabditis bacteriophora. The mean abundance of total EPN community was 28,000ind./m(2) in oak wood and 11,000ind./m(2) in meadow. The seasonal dynamics of entomopathogenic nematodes in both habitats were characterized by high nematode densities in the beginning of the season, followed by a rapid decrease, and then stabilization. EPN abundances did not show any apparent correlation with soil temperature and moisture, but they were negatively correlated with the abundance of suitable insect hosts. Inter- and intraspecific competition for limited nutrients (hosts) probably played a major role in EPN seasonal dynamics. Broad host range of entomopathogenic nematodes in both habitats was predominantly represented by dipteran and coleopteran larvae. Most common hosts belonged to the families Asilidae, Bibionidae, and Empididae (Diptera), as well as Carabidae and Curculionidae (Coleoptera). PMID- 15893762 TI - Effects of menadione and its derivative on cultured cardiomyocytes with mitochondrial disorders. AB - Mitochondrial disorder is characteristic of many myocardial injuries such as endotoxemia, shock, acidosis, ischemia/reperfusion, and others. The goal of possible therapy is to increase ATP production. Derivatives of vitamins K may be a potent electron carrier between various mitochondrial electron-donating and electron-accepting enzyme complexes. We aimed to test the possibility that menadione or its water-soluble derivative AK-135, the newly synthesized analogues of vitamin K1--N-derivatives of 2-methyl-3-aminomethyl 1.4-naphthoquinone, would reduce cardiomyocyte damage after hypoxia or mitochondrial respiratory chain inhibition in culture. Menadione, and more effectively, AK-135, restored the electron flow in defective respiratory chain (hypoxia or rotenone) systems. As was shown in this study, 3 microM of AK-135 restored ATP production after blockade of electron flow through mitochondrial complex I with 5 microM rotenone up to 13.18+/-1.56 vs. 3.21+/-1.12 nmol/mg protein in cells treated with rotenone only. In cultures pretreated with 4 microM dicumarol (DT-diaphorase inhibitor), the protective effect of AK-135 and menadione was abolished completely (1.67+/ 1.43 and 2.97+/-0.57 nmol/mg protein, respectively). Inhibition of mitochondrial oxidative phosphorylation caused an increase in intracellular Ca(2+) levels. Here we have demonstrated restoration of calcium oscillations and cardiomyocyte contractility by menadione and its derivative after blockade of NADH: ubiquinone oxidoreductase with rotenone, and decrease of Ca(2+) overloading during hypoxia. PMID- 15893763 TI - Nuclear-mitochondrial cross-talk in cardiomyocyte T3 signaling: a time-course analysis. AB - Thyroid hormone (TH) induces marked changes in the biochemical and physiological functioning of cardiac muscle affecting its bioenergetics, contractility and structure. Using a time-course analysis of in vitro treatment of neonatal rat cardiomyocytes with triiodothyronine (T3), mitochondrial biogenesis, functional bioenergetics and cardiomyocyte hypertrophic phenotype were assessed. Activity of respiratory complexes II, IV, V and citrate synthase (CS), levels of mitochondrial enzyme subunits (e.g. COXI, COXIV) and nuclear-encoded transcription factors, involved in mitochondrial biogenesis (e.g. PGC-1, mtTFA and PPAR-alpha), were significantly elevated with 72 h T3 treatment. A time course analysis showed an early increase (between 3 and 12 h) in activity and levels of subunits of complex IV and V, mitochondrial Ca2+ accumulation and a late increase (at 72 h) in complex II and CS activities, mitochondrial protein content and mitochondrial respiration. Based on overall protein content and specific peptide levels (e.g. actin or myosin) only mild cardiomyocyte hypertrophy was detected. T3 mediates an early stimulation of enzymes containing mtDNA encoded subunits (e.g. complex IV and V) in contrast to a different regulatory pattern for the entirely nuclear-encoded enzymes (e.g. CS and complex II). T3-regulation was similar in both neonatal and young adult cardiomyocytes (ARCM) but absent in the senescent cardiomyocytes. This model offer an opportunity to study the rapid timing of events involved in myocardial cell signaling, bioenergetics and growth dynamics in a timeframe not available with whole animal studies. PMID- 15893764 TI - Phospholipase C and cAMP-dependent positive inotropic effects of ATP in mouse cardiomyocytes via P2Y11-like receptors. AB - ATP is released as a cotransmitter together with catecholamines from sympathetic nerves. In the heart ATP has been shown to cause a pronounced positive inotropic effect and may also act in synergy with beta-adrenergic agonists to augment cardiomyocyte contractility. The aim of the present study was to investigate the inotropic effects mediated by purinergic P2 receptors using isolated mouse cardiomyocytes. Stable adenine nucleotide analogs were used and the agonist rank order for adenine nucleotide stimulation of the mouse cardiomyocytes was AR C67085>ATPgammaS>2-MeSATP>>>2-MeSADP=0, that fits the agonist profile of the P2Y11 receptor. ATPgammaS induced a positive inotropic response in single mouse cardiomyocytes. The response was similar to that for the beta1 receptor agonist isoproterenol. The most potent response was obtained using AR-C67085, a P2Y11 receptor agonist. This agonist also potentiated contractions in isolated trabecular preparations. The adenylyl cyclase blocker (SQ22563) and phospholipase C (PLC) blocker (U73122) demonstrated that both pathways were required for the inotropic response of AR-C67085. A cAMP enzyme immunoassay confirmed that AR C67085 increased cAMP in the cardiomyocytes. These findings are in agreement with the P2Y11 receptor, coupled both to activation of IP3 and cAMP, being a major receptor for ATP induced inotropy. Analyzing cardiomyocytes from desmin deficient mice, Des-/-, with a congenital cardiomyopathy, we found a lower sensitivity to AR-C67085, suggesting a down-regulation of P2Y11 receptor function in heart failure. The prominent action of the P2Y11 receptor in controling cardiomyocyte contractility and possible alterations in its function during cardiomyopathy may suggest this receptor as a potential therapeutic target. It is possible that agonists for the P2Y11 receptor could be used to improve cardiac output in patients with circulatory shock and that P2Y11 receptor antagonist could be beneficial in patients with congestive heart failure (CHF). PMID- 15893765 TI - Targeted nNOS gene transfer into the cardiac vagus rapidly increases parasympathetic function in the pig. AB - Nitric oxide (NO) derived from neuronal nitric oxide synthase (nNOS) facilitates cardiac vagal neurotransmission and bradycardia in vitro. Here we provide evidence of rapid (within 9 h) protein expression and increased vagal responsiveness in vivo following targeted gene transfer of nNOS into the cardiac vagus of the pig. Right vagi were injected with vector encoding nNOS (Ad.nNOS) or saline, while left vagi received an injection of vector encoding enhanced green fluorescent protein (Ad.eGFP). Enhanced nNOS protein expression was detected exclusively in the right vagus nerve, with no evidence of iNOS expression. This was associated with increased baroreflex sensitivity and greater heart rate responsiveness to right vagal stimulation. In contrast, responsiveness of left vagi, or sham-injected right vagi remained constant over the same time period. Basal heart rate was unchanged following gene transfer, suggesting no change in vagal tone. These results support the pre-/post-ganglionic synapse as a site for NO-mediated facilitation of vagal bradycardia in the pig. In addition they demonstrate in vivo that functional gene expression induced with adenoviral vectors occurs earlier than first thought, and may therefore, provide a novel intervention to acutely modulate the neural control of cardiac excitability. PMID- 15893766 TI - Regulation of RANKL-induced osteoclastic differentiation by vascular cells. AB - Vascular calcification is a regulated process of biomineralization resembling osteogenesis. Many bone-related factors, including resorptive osteoclast-like cells, although in low abundance, have been found in calcified atherosclerotic lesions. The regulatory mechanisms governing them in the vasculature, however, are not clear. Previously, we found that calcifying vascular cells (CVC), a subpopulation of bovine aortic smooth muscle cells (BASMC), undergo osteoblastic differentiation and form mineralized nodules. Since osteoblasts and marrow stromal preosteoblasts regulate osteoclastic differentiation in bone, we hypothesized that vascular cells also regulate differentiation of osteoclastic precursors in the artery wall. Peripheral blood monocytes, which are osteoclast precursors, were co-cultured with CVC or BASMC. Results showed that monocytes co cultured with both of the vascular cells yielded fewer resorption pits than monocytes cultured alone. Furthermore, monocytes co-cultured with CVC had fewer resorption pits than those co-cultured with BASMC. Conditioned media from the vascular cells also inhibited resorptive activity of monocytes suggesting that the inhibitory effect was mediated in part by soluble factors. Compared with BASMC, CVC had lower mRNA expression for osteopontin, which promotes osteoclast attachment, but greater mRNA expression for the soluble inhibitory cytokine, IL 18. Increased osteoclastic differentiation was observed when neutralizing antibody to IL-18 receptor was added to the cultures of preosteoclasts with CVC conditioned media. Osteoprotegerin, another osteoclast inhibitory cytokine, was expressed at similar levels in both cultures. These results suggest that vascular cells inhibit osteoclastic differentiation, and that CVC have greater inhibitory effects than BASMC. PMID- 15893767 TI - Two novel targeting peptide degrading proteases, PrePs, in mitochondria and chloroplasts, so similar and still different. AB - Two novel metalloproteases from Arabidopsis thaliana, termed AtPrePI and AtPrePII, were recently identified and shown to degrade targeting peptides in mitochondria and chloroplasts using an ambiguous targeting peptide. AtPrePI and AtPrePII are classified as dually targeted proteins as they are targeted to both mitochondria and chloroplasts. Both proteases harbour an inverted metal binding motif and belong to the pitrilysin subfamily A. Here we have investigated the subsite specificity of AtPrePI and AtPrePII by studying their proteolytic activity against the mitochondrial F(1)beta pre-sequence, peptides derived from the F(1)beta pre-sequence as well as non-mitochondrial peptides and proteins. The degradation products were analysed, identified by MALDI-TOF spectrometry and superimposed on the 3D structure of the F(1)beta pre-sequence. AtPrePI and AtPrePII cleaved peptides that are in the range of 10 to 65 amino acid residues, whereas folded or longer unfolded peptides and small proteins were not degraded. Both proteases showed preference for basic amino acids in the P(1) position and small, uncharged amino acids or serine residues in the P'(1) position. Interestingly, both AtPrePI and AtPrePII cleaved almost exclusively towards the ends of the alpha-helical elements of the F(1)beta pre-sequence. However, AtPrePI showed a preference for the N-terminal amphiphilic alpha-helix and positively charged amino acid residues and degraded the F(1)beta pre-sequence into 10-16 amino acid fragments, whereas AtPrePII did not show any positional preference and degraded the F(1)beta pre-sequence into 10-23 amino acid fragments. In conclusion, despite the high sequence identity between AtPrePI and AtPrePII and similarities in cleavage specificities, cleavage site recognition differs for both proteases and is context and structure dependent. PMID- 15893768 TI - Crystal structures of the tricorn interacting factor F3 from Thermoplasma acidophilum, a zinc aminopeptidase in three different conformations. AB - The tricorn interacting factor F3 is an 89 kDa zinc aminopeptidase from the archaeon Thermoplasma acidophilum. Together with the tricorn interacting factors F1 and F2, F3 degrades the tricorn protease products and thus completes the proteasomal degradation pathway by generating free amino acids. Here, we present the crystal structures of F3 in three different conformations at 2.3 A resolution. The zinc aminopeptidase is composed of four domains: an N-terminal saddle-like beta-structure domain; a thermolysin-like catalytic domain; a small barrel-like beta-structure domain; and an alpha-helical C-terminal domain, the latter forming a deep cavity at the active site. Three crystal forms provide snapshots of the molecular dynamics of F3 where the C-terminal domain can adapt to form an open, an intermediate and a nearly closed cavity, respectively. With the conserved Zn(2+)-binding motifs HEXXH and NEXFA as well as the N-terminal substrate-anchoring glutamate residues, F3 together with the leukotriene A4 hydrolase, represents a novel gluzincin subfamily of aminoproteases. We discuss the functional implications of these structures with respect to the underlying catalytic mechanism, substrate recognition and processing, and possible component interactions. PMID- 15893769 TI - Tertiary interactions stabilise the C-terminal region of human glutathione transferase A1-1: a crystallographic and calorimetric study. AB - The C-terminal region in class Alpha glutathione transferase A1-1 (GSTA1-1), which forms an amphipathic alpha-helix (helix 9), is known to contribute to the catalytic and non-substrate ligand-binding functions of the enzyme. The region in the apo protein is proposed to be disordered which, upon ligand binding at the active-site, becomes structured and localised. Because Ile219 plays a pivotal role in the stability and localisation of the region, the role of tertiary interactions mediated by Ile219 in determining the conformation and dynamics of the C-terminal region were studied. Ligand-binding microcalorimetric and X-ray structural data were obtained to characterise ligand binding at the active-site and the associated localisation of the C-terminal region. In the crystal structure of the I219A hGSTA1-1.S-hexylglutathione complex, the C-terminal region of one chain is mobile and not observed (unresolved electron density), whereas the corresponding region of the other chain is localised and structured as a result of crystal packing interactions. In solution, the mutant C-terminal region of both chains in the complex is mobile and delocalised resulting in a hydrated, less hydrophobic active-site and a reduction in the affinity of the protein for S hexylglutathione. Complete dehydration of the active-site, important for maintaining the highly reactive thiolate form of glutathione, requires the binding of ligands and the subsequent localisation of the C-terminal region. Thermodynamic data demonstrate that the mobile C-terminal region in apo hGSTA1-1 is structured and does not undergo ligand-induced folding. Its close proximity to the surface of the wild-type protein is indicated by the concurrence between the observed heat capacity change of complex formation and the type and amount of surface area that becomes buried at the ligand-protein interface when the C terminal region in the apo protein assumes the same localised structure as that observed in the wild-type complex. PMID- 15893770 TI - Bacterial repression loops require enhanced DNA flexibility. AB - The Escherichia coli lac operon provides a classic paradigm for understanding regulation of gene transcription. It is now appreciated that lac promoter repression involves cooperative binding of the bidentate lac repressor tetramer to pairs of lac operators via DNA looping. We have adapted components of this system to create an artificial assay of DNA flexibility in E.coli. This approach allows for systematic study of endogenous and exogenous proteins as architectural factors that enhance apparent DNA flexibility in vivo. We show that inducer binding does not completely remove repression loops but it does alter their geometries. Deletion of the E.coli HU protein drastically destabilizes small repression loops, an effect that can be partially overcome by expression of a heterologous mammalian HMG protein. These results emphasize that the inherent torsional inflexibility of DNA restrains looping and must be modulated in vivo. PMID- 15893771 TI - Expression of STAT3 and SOCS3 in pancreatic acinar cells. AB - BACKGROUND: The signal transducer and activator of transcription (STAT) and suppressor of cytokine signaling 3 (SOCS3) pathways are involved in organ inflammation. In the pancreas, however, the STAT3 and SOCS3 response to inflammatory stimuli is unknown. Therefore, we hypothesized that mRNA expression for these signaling proteins would be induced in response to pro-inflammatory mediators TNFalpha and LPS. Because activation of STAT3 and SOCS3 is also linked to cytokine stimulation, we tested TNFalpha and LPS, either alone or in combination with IL-1beta or IL-6 in an in vitro model using rat pancreatic acinar cells. METHODS: Rat pancreatic acinar cells (AR42J) were treated with combinations of LPS (10 microg/ml) or TNFalpha (10, 100, or 200 ng/ml) in the presence or absence of IL-1beta or IL-6 for 15, 30, 45, 60, 180, or 360 min. At each time point, total RNA was purified and analyzed via RT-PCR for STAT3 and SOCS3 mRNA expression. RESULTS: LPS and TNFalpha activated STAT3 and SOCS3 in pancreatic acinar cells. STAT3 mRNA expression was significantly (P < 0.01) increased above controls without further stimulation by IL-6 or IL1-beta. Significant increases in SOCS3 expression were observed with LPS + IL-6 at 30, 45, 60, 180, min (P < 0.05). SOCS3 mRNA expression with LPS + IL-1beta treatment was maximal by 1 h (P < 0.05). Enhanced STAT3 expression was evident by 3 h with TNFalpha alone (P < 0.01). The addition of cytokines kept STAT3 mRNA levels high. At 200 ng/ml TNFalpha, SOCS3 mRNA levels were increased, but significantly reduced in the presence of IL-1beta or IL-6 (P < 0.05). CONCLUSIONS: We have shown that LPS and TNFalpha are potent mediators of STAT3 and SOCS3 expression in the pancreas and that the cytokines IL-6 and IL-1beta are indirectly involved in the signaling pathway. Alteration of the STAT pathway should be explored as a therapeutic target for treatment of pancreatic inflammation. PMID- 15893772 TI - Puberty installation and adrenergic response of seminal vesicle from rats exposed prenatally to hydrocortisone. AB - We investigated the effects of hydrocortisone during the prenatal period and its repercussion on puberty installation and adrenergic response of seminal vesicle in adult rats. The efficacy of the hydrocortisone treatment in reducing adrenal wet weight immediately after delivery in both the treated mothers and respective pups at birth may indicate impairment of the hypothalamus--pituitary--adrenal axis. This parameter was unchanged in the adult phase of these descendants, suggesting recuperation of this axis. In addition, the treatment with hydrocortisone delayed the age of puberty installation, probably by absence of both physiologic production and liberation of luteinizing hormone and testosterone. Despite the significant reduction in testosterone level as well as of wet weights of both vas deferens and testis in the adult phase, no difference was observed in the sensitivity of the seminal vesicle to the studied sympathetic agonist. However, the observed reduction in contractile response of the seminal vesicle may be a consequence of contractile-system damage in this organ. It is possible that this alteration may cause a reduction in the amount of vesicular secretion so important in the process of ejaculation. In conclusion, these results suggest that administration of hydrocortisone in late prenatal life did not influence the hypothalamus--pituitary--adrenal axis in adult life, although it altered the hypothalamus--pituitary--gonadal axis, and reduced testosterone production starting at puberty, as a consequence of an incomplete masculinization of the hypothalamus plus a reduction in the contractile response of the seminal vesicle. PMID- 15893773 TI - Adiponectin-mediated stimulation of AMP-activated protein kinase (AMPK) in pancreatic beta cells. AB - The adipocyte-derived hormone adiponectin was recently shown to stimulate glucose utilization and to increase fatty acid oxidation in liver and muscle. The effects were ascribed to adiponectin-receptor mediated activation of the key metabolic regulator AMP-activated protein kinase (AMPK). In pancreatic beta cells, AMPK activation is known to affect cellular function. We therefore investigated a possible adiponectin-induced activation of AMPK in beta cells. RT-PCR analysis confirmed the expression of adiponectin receptor subtypes 1 and 2 in rat beta cells and showed their expression in insulin-secreting MIN6 cells. Culture with physiological concentrations (2.5 microg/ml) of globular adiponectin was found to increase the phosphorylation of both AMPK and acetylcoA carboxylase (ACC) in these cell types. Like the pharmacological AMPK activator 5-amino-imidazole-4 carboxamide-riboside (AICAR), adiponectin activated AMPK in beta cells and MIN6 cells. In short-term incubations of MIN6 cells with either adiponectin (2.5 microg/ml) or AICAR (1 mM), the flux of glucose-carbon to acyl CoA/cholesterol biosynthetic intermediates was reduced. We conclude that adiponectin induces an activation of AMPK in beta cells, which inhibits their cataplerosis of glucose carbon to lipids. PMID- 15893774 TI - Ketone bodies affect the enzymatic activity of macrophage migration inhibitory factor. AB - Macrophage migration inhibitory factor (MIF), a long known proinflammatory cytokine exhibits perplexing enzymatic activities: tautomeric conversion of D dopachrome and phenylpyruvate. Whether these catalytic activities bear functional relevance regarding MIF's multifaceted roles is under current scrutiny. Nevertheless, intense search has already started for pharmacological agents that target MIF's tautomerase activity. We have probed several antiinflammatory compounds against keto--enol (enolase) and enol--keto (ketonase) conversion of phenylpyruvate by MIF with spectrophotometry. We have identified acidic CH groups as markers of inhibitor potency toward MIF phenylpyruvate tautomerase. Among simple model molecules with strong acidic CH groups we found acetylacetone the best inhibitor particularly against the ketonase activity. Ketones of physiological importance - ketone bodies - also feature acidic CH groups and have been reported to exert certain anti-inflammatory effects. In this paper we report that ketone bodies inhibit preferentially the ketonase activity of MIF in vitro. Future studies should address whether such an interaction might operate in vivo and delineate its possible relevance concerning cytokine and non-cytokine roles of MIF. PMID- 15893775 TI - The effect of local nitric oxide synthase inhibition on the diameter of pulpal arteriole in dental bond material-induced vasodilation in rat. AB - Local application of dental bond materials can cause pulpal vasodilation and hyperemia. Such local hemodynamic changes may be mediated by alterations in the levels of locally generated nitric oxide (NO). In different species systemic administration of NO synthase inhibitors leads to a decrease in pulpal blood flow. In contrast, the local administration of these inhibitors has not been tested yet. Therefore, the effect of locally blocked NO synthase on the internal diameter of rat pulpal arterioles under basal conditions and immediately after dental bond material application was studied by using vitalmicroscopic technique. The NO synthase blocker (L-NAME) was locally administered on a thinned dentine layer of the left lower incisor. L-NAME reduced the diameter of the pulpal arteriole both in basal and after bond material-induced hyperaemic conditions. These data suggest that the local formation of NO may have a significant role in the acute vasodilation induced by bond material application and also in maintenance of basal pulpal arteriolar tone. PMID- 15893776 TI - Unilateral basal ganglia damage causes contralesional force control deficits: a case study. AB - When grasping to lift an object, the grip force is usually scaled to the mass of the object. However, it has been shown that lifting objects of different sizes but equal masses results in the generation of higher forces for larger compared to smaller objects. The objective of this study was to investigate whether a similar effect is present in an individual (RI) with a unilateral lesion to the basal ganglia (BG). It was hypothesized that if the BG have an influence on the use of visual information in updating of the internal model used to anticipate the forces required for grasping, damage to these structures should result in the inability of RI's contralesional hand to anticipate object mass based on size cues. To test this hypothesis three objects of equal mass but different sizes were grasped and lifted by RI and six control individuals. The forces that were generated during these lifts were quantified. The controls showed the expected increases in peak grip force as object size increased. RI showed no effect of object size for his contralesional hand, but did show force scaling with his ipsilesional hand. In conclusion, RI's BG damage affected the on-line control of grip forces and the inability to integrate visual and tactile information in the programming of finger forces. PMID- 15893777 TI - Peripersonal spatial attention in children with spina bifida: associations between horizontal and vertical line bisection and congenital malformations of the corpus callosum, midbrain, and posterior cortex. AB - Horizontal and vertical line bisection was studied in 129 children and adolescents between 8 and 19 years of age, one group (n=32) of typically developing controls and one group (n=97) with spina bifida (SBM), a neurodevelopmental disorder associated with dysmorphology of the corpus callosum, posterior cortex, and midbrain. For each participant, structural brain MRIs were analyzed qualitatively to identify beaking of the midbrain tectum and corpus callosum agenesis and hypoplasia and quantitatively by segmentation and volumetric analyses of regional cortical white and gray matter. Each group showed the line length effect, whereby greater estimation errors are made with longer lines. The group with SBM differed from controls in terms of both accuracy and variability of line bisection. Children with SBM showed pseudoneglect, attending more than controls to left hemispace. The extent of rightward bisection bias was unrelated to right posterior brain volumes, although an intact corpus callosum during development moderated and normalized the exaggerated leftward line bisection bias. More children with SBM than controls attended to inferior hemispace. A normal midbrain and greater posterior cortex volume during development moderated and normalized the downward bias. Children with SBM showed more intra-subject variability than controls. Line bisection in children with SBM reflects three deficits: an exaggerated attentional bias to left hemispace, an abnormal attentional bias to inferior hemispace; and a larger zone of subjective uncertainty in bisection judgments. PMID- 15893778 TI - Feeding response to melanocortin agonist predicts preference for and obesity from a high-fat diet. AB - Overconsumption and increased selection of high fat (HF) foods contribute to the development of common obesity. Because the hypothalamic melanocortin (MC) system plays an integral role in the regulation of food intake and dietary choice, we tested the hypothesis that proneness (-P) or resistance (-R) to dietary-induced obesity (DIO) may be due to differences in MC function. We found that prior to developing obesity and while still maintained on chow, acute, central administration of MTII, an MC agonist, produced a greater anorectic response in DIO-P rats than in DIO-R rats. However, after only 5 days of exclusive HF feeding, the DIO-R rats had significantly greater suppression of intake after MTII treatment than they did when maintained on chow. In addition, the DIO-P rats were much less responsive to MTII treatment than the DIO-R rats after only 5 days of the HF diet. In fact, MTII-induced anorexia during HF feeding correlated negatively with body weight gained on the HF diet. These results suggest that the voluntary decrease of HF feeding in DIO-R rats may be mediated by increased endogenous MC signaling, a signal likely compromised in DIO-P rats. Differences in MC regulation may also explain the observed preference for HF over a lower fat food choice in DIO-P rats. Finally, the results indicate that responses to exogenous MC challenge can be used to predict proneness or resistance to DIO. PMID- 15893779 TI - Four tetranortriterpenoids from the stem bark of Khaya anthotheca. AB - Eight limonoids, anthothecanolide (1), 3-O-acetylanthothecanolide (2), 2,3-di-O acetylanthothecanolide (3), 6R,8alpha-dihydroxycarapin (4), 3beta-acetoxy-3-deoxo 6R-hydroxycarapin (5), methyl angolensate, methyl 6-hydroxyangolensate and khayalactone together with sitosterol glucoside, have been isolated from the extracts of the stem bark of Khaya anthotheca. Compounds 1-4 are described for the first time. Their structures were established by analysis of the high-field NMR and MS data. The structure of compound 4 was confirmed by a single crystal X ray structure analysis. PMID- 15893780 TI - Predictors of smoking cessation during pregnancy among the women of Yamato and Ayase municipalities in Japan. AB - Cigarette smoking during pregnancy affects the unborn fetus. This study aimed to investigate: (1) the smoking status of pregnant women before, during and after pregnancy in the Yamato and Ayase municipalities, Kanagawa, Japan; (2) the characteristics that differentiate successful spontaneous smoking quitters from continuous smokers during pregnancy; and (3) the awareness of risks of smoking that may influence smoking cessation during pregnancy. A community-based, cross sectional survey of 420 postpartum women, who delivered their babies from July 2002 to October 2002, was performed. Out of 420 respondents, 275 (65.4%) did not smoke before and during pregnancy. Of the 145 women who smoked before pregnancy, 101 (69.3%) quit successfully while pregnant. Smoking prevalence during pregnancy was 10.4%. Out of 101 successful quitters during pregnancy, 22 (21.8%) women resumed smoking after childbirth. A multiple regression analysis showed that the independent predictors of smoking cessation during pregnancy were primiparous women and women living with non-smokers in the household. Although maternal age predicted smoking cessation in a simple regression analysis, this predictive value was lost in the multiple analysis. A simple analysis showed no significant association between the awareness of smoking risks and smoking cessation during pregnancy. It is suggested that Japanese smokers are more likely to quit while pregnant than women in other countries. In the future, smoking cessation programmes should be designed to help multiparous women and women living with smokers to give up smoking. PMID- 15893781 TI - Treatable factors associated with severe anaemia in adults admitted to medical wards in Blantyre, Malawi, an area of high HIV seroprevalence. AB - Severe anaemia is a common presentation in non-pregnant adults admitted to hospital in southern Africa. Standard syndromic treatment based on data from the pre-HIV era is for iron deficiency, worms and malaria. We prospectively investigated 105 adults admitted consecutively to medical wards with haemoglobin < 7 g/dl. Those with acute blood loss were excluded. Patients were investigated for possible parasitic, bacterial, mycobacterial and nutritional causes of anaemia, including bone marrow aspiration, to identify potentially treatable causes. Seventy-nine per cent of patients were HIV-positive. One-third of patients had tuberculosis, which was diagnosed only by bone marrow culture in 8% of HIV-positive patients. In 21% of individuals bacteria were cultured, with non typhi salmonella predominating and Streptococcus pneumoniae rare. Iron deficiency, hookworm infection and malaria were not common in HIV-positive anaemic adults, although heavy hookworm infections were found in 6 (27%) of the 22 HIV-negative anaemic adults. In conclusion, conventional treatment for severe anaemia in adults is not appropriate in an area of high HIV prevalence. Occult mycobacterial disease and bacteraemia are common, but iron deficiency is not common in HIV-positive patients. In addition to iron supplements, management of severe anaemia should include investigation for tuberculosis, and consideration of antibiotics active against enterobacteria. PMID- 15893782 TI - Immunomodulatory effects of the fungicide Mancozeb in agricultural workers. AB - Available data suggest that ethylenebisdithiocarbamates (EBDCs) may have immunomodulatory effects. This study aimed to investigate the immunological profile of farmers exposed to Mancozeb, an EBDC fungicide, through the determination of several serum, cellular, and functional immune parameters. Twenty-six healthy subjects entered the study, 13 vineyards exposed to Mancozeb and 13 unexposed controls. Exposure was assessed through the determination of ethylentiourea (ETU) in urine. Complete and differential blood count, serum immunoglobulins, complement fractions, autoantibodies, lymphocyte subpopulations, proliferative response to mitogens, natural killer (NK) activity, and cytokine production were measured. Post-exposure samples showed ETU urine concentration significantly higher than pre-exposure and control groups. A significant increase in CD19+ cells, both percentage and absolute number, and a significant decrease in the percentage of CD25+ cells were found in post-exposure samples compared to controls. A statistically significant increase in the proliferative response to phorbol myristate acetate plus ionomycin (PMA + ionomycin) was observed in the post-exposure group compared to controls and baseline, while a significant reduction in LPS-induced TNF-alpha release in post-exposure samples was observed. Overall, our results suggest that low-level exposure to Mancozeb has slight immunomodulatory effects, and point out a method adequate to reveal immune modifications in workers occupationally exposed to potential immunotoxic compounds, based on a whole blood assay. PMID- 15893783 TI - The fusion protein of wild-type canine distemper virus is a major determinant of persistent infection. AB - The wild-type A75/17 canine distemper virus (CDV) strain induces a persistent infection in the central nervous system but infects cell lines very inefficiently. In contrast, the genetically more distant Onderstepoort CDV vaccine strain (OP-CDV) induces extensive syncytia formation. Here, we investigated the roles of wild-type fusion (F(WT)) and attachment (H(WT)) proteins in Vero cells expressing, or not, the canine SLAM receptor by transfection experiments and by studying recombinants viruses expressing different combinations of wild-type and OP-CDV glycoproteins. We show that low fusogenicity is not due to a defect of the envelope proteins to reach the cell surface and that H(WT) determines persistent infection in a receptor-dependent manner, emphasizing the role of SLAM as a potent enhancer of fusogenicity. However, importantly, F(WT) reduced cell-to-cell fusion independently of the cell surface receptor, thus demonstrating that the fusion protein of the neurovirulent A75/17-CDV strain plays a key role in determining persistent infection. PMID- 15893784 TI - Effects of electrical microstimulation in monkey frontal eye field on saccades to remembered targets. AB - Spatially selective delay activity in the frontal eye field (FEF) is hypothesized to be part of a mechanism for the transformation of visual signals into instructions for voluntary movements. To understand the linkage between FEF activity and eye movement planning, we recorded neuronal responses of FEF neurons while monkeys performed a memory-saccade task. We then electrically stimulated the same sites during the memory-delay epoch of the task. The stimulation currents used were subthreshold for evoking saccades during a gap-fixation task. Microstimulation resulted in changes in the spatial and temporal components of saccade parameters: an increase in latency, and a shift in amplitude and direction. We performed a vector analysis to determine the relative influence of the visual cue and electrical stimulus on the memory-saccade. In general, the memory-saccade was strongly weighted toward the visual cue direction, yet the electrical stimulus introduced a consistent bias away from the receptive/movement field of the stimulation site. The effects of sub-threshold stimulation were consistent with a combination of vector subtraction and averaging, but not with vector summation. Vector subtraction may play a role in spatial updating of movement plans for memory-guided saccades when eye position changes during the memory period. PMID- 15893785 TI - Crowding degrades saccadic search performance. AB - The identity of a target is more difficult to acquire when it is surrounded by distracters. The purpose of the present experiments was to investigate the implications of this crowding phenomenon for performance and eye movements in a real-life task as search with eye movements. The participants searched for a target in a one dimensional search strip. Above and below this search strip additional elements were added. In three conditions, the similarity of these mask elements to the search elements was varied. The spatial extent of crowding is known to increase with target-mask similarity [Nazir, T. A. (1992). Effects of lateral masking and spatial precueing on gap-resolution in central and peripheral vision. Vision Research, 32, 771-777, Kooi, F. L., Toet, A., Tripathy, S. P., & Levi, D. M. (1994). The effect of similarity and duration on spatial interaction in peripheral vision. Spatial Vision, 8(2), 255-279]. One condition did not contain masks. In a visibility experiment, we firstly validated this crowding manipulation. In the search experiment, we subsequently found that with increasing crowding search times were up to 76% longer. Eye movements were also affected. The number of fixations and fixation duration increased and saccade amplitude decreased with increasing crowding. We conclude that in order to understand eye movements in (everyday) tasks that require active exploration of the visual scene, crowding should be taken into account. PMID- 15893786 TI - Interactions between binocular rivalry and Gestalt formation. AB - A question raised a long time ago in binocular rivalry research is whether the phenomenon of binocular rivalry is purely determined by local stimulus properties or that global stimulus properties also play a role. More specifically: do coherent features in a stimulus influence rivalrous behavior? After decades of underexposure of the subject, recently this question seemed to be answered in the affirmative. This paper presents additional evidence for an influence of coherent features. In an experiment in which eye movements cannot bias conclusions it is demonstrated that Gestalt formation influences binocular rivalry positively, i.e., stronger Gestalts have longer total dominance times. Gestalt formation appears to intervene in the states of dominance ("what"), not directly in the dominance durations ("how long"). This generates questions about the nature of interactions between binocular rivalry and Gestalt formation. Gestalt formation seems to be fed by signals that are generated after binocular convergence and only leaves its mark on binocular rivalry by feedback to monocular channels, a conclusion which has been drawn before by Alais and Blake [Alais, D., & Blake, R. (1998). Interaction between global motion and local binocular rivalry. Vision research 38, 637-644]. PMID- 15893787 TI - Chemical and biochemical generation of carbohydrates from lignocellulose feedstock (Lupinus nootkatensis)--quantification of glucose. AB - Different chemical and enzymatic methods were applied for the hydrolysis of main stems from Lupinus nootkatensis (harvest November 2002). The whole process (all steps) is based on the lignocellulose-feedstock biorefinery regime. The acid hydrolysis of L. was performed with concentrated hydrochloric acid; advantages in this process are exothermic hydrolysis and the possibility of acid recovery. Enzymatic hydrolysis achieved high yields of fermentable carbohydrates (regarding to input cellulose) with high selectivity. However, this way requires the generation of cellulose from L. by chemical pulping. Monosaccharide derivatives thus obtained were identified by their GC retention times and the corresponding MS fragmentation. Hexamethyldisilazane was used as derivatization reagent to prepare the trimethylsilyl derivatives of the carbohydrates and of the degradations products of cellulose from the different fractions. The glucose content was quantified by GC peak integration with respect to an internal standard. PMID- 15893788 TI - Chemical characterization of extractable water soluble matter associated with PM10 from Mexico City during 2000. AB - We report the chemical composition of PM10-associated water-soluble species in Mexico City during the second semester of 2000. PM10 samples were collected at four ambient air quality monitoring sites in Mexico City. We determined soluble ions (chloride, nitrate, sulfate, ammonium, sodium, potassium), ionizable transition metals (Zn, Fe, Ti, Pb, Mn, V, Ni, Cr, Cu) and soluble protein. The higher PM(10) levels were observed in Xalostoc (45-174 microg m(-3)) and the lowest in Pedregal (19-54 microg m(-3)). The highest SO2 average concentrations were observed in Tlalnepantla, NO2 in Merced and O3 and NO(x) in Pedregal. The concentration range of soluble sulfate was 6.7-7.9 and 19-25.5 microg m(-3) for ammonium, and 14.8-29.19 for soluble V and 3.2-7.7 ng m(-3) for Ni, suggesting a higher contribution of combustion sources. PM-associated soluble protein levels varied between 0.038 and 0.169 mg m(-3), representing a readily inhalable constituent that could contribute to adverse outcomes. The higher levels for most parameters studied were observed during the cold dry season, particularly in December. A richer content of soluble metals was observed when they were expressed by mass/mass units rather than by air volume units. Significant correlations between Ni-V, Ni-SO4(-2), V-SO4(-2), V-SO2, Ni-SO2 suggest the same type of emission source. The variable soluble metal and ion concentrations were strongly influenced by the seasonal meteoclimatic conditions and the differential contribution of emission sources. Our data support the idea that PM10 mass concentration by itself does not provide a clear understanding of a local PM air pollution problem. PMID- 15893789 TI - Distribution and partition of heavy metals in surface and sub-surface sediments of Naples city port. AB - The aim of the study was to delineate the extent of heavy metal pollution in the marine sediments within the port of Naples. Total metal contents from twenty surface sediments were compared with those from a long sediment core representing the natural geochemical baseline. Enrichment factors were computed for each metal and for each site in order to assess the polluting metals and the degree of pollution at each site. Results revealed that heavy metal pollution is mainly localized in the port area devoted to shipbuilding activities and in the south east sector, which is under the influence of petroleum refineries. Data from sequential extractions indicate that metals from anthropogenic sources are mainly Cd, Zn, Cr and Cu and are potentially more mobile than those inherited from geological parent material. PMID- 15893790 TI - LP/LIF study of the formation and consumption of mercury (I) chloride: kinetics of mercury chlorination. AB - The laser photolysis/laser induced fluorescence (LP/LIF) technique has been applied to studies of gas-phase mercury (Hg) chlorination. Mercury (I) chloride (HgCl) was been detected via LIF at 272 nm from reactions of elemental Hg with Cl atoms generated from the 193 nm photolysis of carbon tetrachloride. While the formation of HgCl was too fast to be observed on millisecond time scales, the kinetics of the consumption of HgCl have been determined at temperatures characteristic of post-combustion conditions. Rate coefficients and Arrhenius parameters for the reaction of HgCl with Cl2, HCl and Cl atoms were determined. The reaction of HgCl with Cl2 was the fastest reaction studied, while the reaction of HgCl with HCl was the only reaction to show any measurable temperature dependence. Estimates of the rate coefficient for the reaction Hg + Cl --> HgCl were determined using a modeling approach. Comparisons of these new measurements with model predictions are discussed. PMID- 15893791 TI - Size- and density-distributions and sources of polycyclic aromatic hydrocarbons in urban road dust. AB - Polycyclic aromatic hydrocarbons (PAHs) present in size- and density-fractionated road dust were measured to identify the important fractions in urban runoff and to analyse their sources. Road dust was collected from a residential area (Shakujii) and a heavy traffic area (Hongo Street). The sampling of road dust from the residential area was conducted twice in different seasons (autumn and winter). The collected road dust was separated into three or four size-fractions and further fractionated into light (<1.7 g/cm3) and heavy (>1.7 g/cm3) fractions by using cesium chloride solution. Light particles constituted only 4.0+/-1.4%, 0.69+/-0.03% and 3.4+/-1.0% of the road dust by weight for Shakujii (November), Shakujii (February) and Hongo Street, respectively but contained 28+/-10%, 33+/ 3% and 44+/-8% of the total PAHs, respectively. The PAH contents in the light fractions were 1-2 orders of magnitude higher than those in the heavy fractions. In the light fractions, the 12PAH contents in February were significantly higher than the 12PAH contents in November (P<0.01), whereas in the heavy fractions, no significant difference was found (P>0.05). Cluster analysis revealed that there was a significant difference in the PAH profiles between locations rather than between size-fractions, density-fractions and sampling times. Multiple regression analysis indicated that asphalt/pavement was the major source of Shakujii road dust, and that tyre and diesel vehicle exhaust were the major sources of finer and coarser fractions collected from Hongo Street road dust, respectively. PMID- 15893792 TI - Transport and fate of dieldrin in poplar and willow trees analyzed by SPME. AB - Dieldrin is a hydrophobic organochlorine insecticide that is persistent in the environment. The fate and transport of dieldrin in trees is important both in the context of potential remediation, as well as food chain impacts through dieldrin transport to shoots and leaves. Experiments were conducted to measure the degree of dieldrin partitioning to plant tissue and the potential for biodegradation of dieldrin in the microbe rich tree rhizosphere. Dieldrin was analyzed in water and plant tissue using headspace solid-phase microextraction (SPME) coupled with gas chromatography. Poplar and willow saplings planted in soil and watered with 10 microgl(-1) dieldrin for up to 9 months showed no adverse effects due to dieldrin exposure and no dieldrin was observed in plant shoots with a method detection limit (MDL) of 7 ngg(-1). One-week hydroponic tests of poplar saplings exposed to aqueous dieldrin also showed no detection of dieldrin in shoots, with an average of 66% of the dieldrin partitioned to the plant roots and an overall mass balance recovery of 76% in the plant-water system. The root concentration factor (RCF) was found to be 30+/-3 ml water g(-1) root. Biodegradation of dieldrin was not observed in an aqueous batch bioreactor containing 8 microgl(-1) dieldrin, nutrients and bacteria from the root zone of a poplar sapling that had been exposed to dieldrin for 9 months. These results show that planting trees is likely to be safe and potentially useful at sites containing low-levels of dieldrin in groundwater. PMID- 15893793 TI - Distribution of endocrine disruptors in the Llobregat River basin (Catalonia, NE Spain). AB - The Llobregat basin is a Mediterranean fluvial system with major agricultural, urban and industrial impacts. We combined chemical quantification by liquid chromatography-mass spectrometry with electrospray interface (LC-ESI-MS) and the recombinant yeast assay (RYA) assays to estimate the loads of endocrine disrupting compounds (EDCs) along the basin. Chemical analysis revealed maximum concentrations (at microg l(-1) level) of alkylphenols at the lower course of the Llobregat River, which correlated with high levels of estrogenic activity detected by RYA. Analysis by RYA and LC-ESI-MS of influent and effluent waters from four sewage treatment plants (STP) discharging into the basin showed the removal of 80-95% of EDCs by STP treatment. Chemical analysis data and RYA data showed a quasi-linear correlation, demonstrating the complementariness of both methods. Our data suggest that the concentrations of the analysed compounds were enough to explain the total estrogenicity of water and STP samples from the Llobregat basin. PMID- 15893794 TI - Congener profiles of PCBs and PCDD/Fs in Yucheng victims fifteen years after exposure to toxic rice-bran oils and their implications for epidemiologic studies. AB - In 1978-1979, a mass poisoning occurred in central Taiwan from rice-bran oil contaminated by heat-degraded PCBs was later called the Yucheng (oil disease in Chinese). Only a few studies have so far investigated the levels of specific polychlorinated biphenyl (PCB) or polychlorinated dibenzodioxin/furan (PCDD/F) congeners in the Yucheng victims. This study aimed to investigate the serum residual levels of thirty-three PCBs and seventeen 2,3,7,8-substituted PCDD/F congeners in the Yucheng victims 15 years after the exposure. Forty-one blood samples were collected from individual Yucheng victims in 1994-1995. The mean levels of total 33 PCBs and 17 PCDD/Fs were 2468 ng/g lipid (13.3 ng/g sample) and 6550 pg/g lipid (30.9 pg/g sample) respectively. The higher levels were found in PCBs #99, #138, #153, #156, #170, #179, and #180 among 33 PCB congeners, while 2,3,4,7,8-PeCDF, 1,2,3,4,7,8-HxCDF, and OCDD had the higher concentrations among 17 PCDD/F congeners. The total TEQ was contributed in decreasing order by 10 PCDFs (44%), three non-ortho-PCBs (24%), six mono-ortho-PCBs (20%), and seven PCDDs (12%). The mean total PCB levels and TEQ value of the 17 PCDD/Fs in the Yucheng victims 15 years after the toxic exposure were still 9 and 46 times higher than those in the general population in Taiwan. Principle component analysis (PCA) indicated that seven PCB congeners, PCBs #99, #138, #153, #156, #170, #179, and #180, accounted for 73% of the total variances in PCBs. On the other hand, six PCDD/F congeners, 2,3,4,7,8-PeCDF, 1,2,3,4,7,8-HxCDF, 1,2,3,6,7,8 HxCDF, 1,2,3,6,7,8-HxCDD, 1,2,3,4,6,7,8-HpCDF, and OCDD, accounted for 97% of the total variances in PCDD/Fs. In addition, PCA revealed that at least three characteristic patterns of congener profiles for PCBs were observed among the Yucheng victims. Similar trend was also observed for PCDD/Fs. These patterns may reflect distinctive exposure scenarios and/or different metabolizing capabilities among the Yucheng victims. We suggest that these patterns, in contrast to total PCB and PCDD/F levels, may be valuable for the future epidemiologic studies when linking exposure with specific health effect. PMID- 15893795 TI - Testing VOC emission measurement techniques in wood-coating industrial processes and developing a cost-effective measurement methodology. AB - Availability of reliable emission measurements of concentrated volatile organic compounds (VOCs) bear great significance in facilitating the selection of a feasible emission abatement technique. There are numerous methods, which can be used to measure VOC emissions, however, there is no single method that would allow sampling of the whole range of volatile organics. In addition, research efforts are usually directed to the development of measuring VOCs in diluted concentrations. Therefore, there is a need for a novel measurement method, which can give reliable results while entailing simple operations and low costs. This paper represents a development effort of finding a reliable measurement procedure. A methodology is proposed and used to measure solvent emissions from coating processes. PMID- 15893796 TI - Characterization of trace metal particles deposited on some deciduous tree leaves in an urban area. AB - In 1996 and 1997 horse chestnut (Aesculus hippocastanum L.) and Turkish hazel (Corulys colurna L.) leaves were sampled at 2 m height in the Belgrade Botanic Garden, located in an urban area with heavy traffic. Using a scanning electron microscope with an energy dispersive X-ray spectroscopy (SEM-EDAX), the size, size distribution, morphology and chemical composition of individual particles were examined on the adaxial and abaxial surfaces of leaf discs of both species. The majority of particles observed on leaves belonged to a class of fine particles (D < 2 microm). Morphological and chemical composition indicated that the most abundant particles were soot and dust with minor constituents such as Pb, Zn, Ni, V, Cd, Ti, As and Cu. Using an electrochemical technique (DPASV), it was possible to measure trace metal concentrations (Pb, Cu, Zn) in a water soluble fraction of deposits on each single leaf. Trace metal contents in the leaf deposits, increased during the vegetation period for both species and were considerably higher in A. hippocastanum due to different epidermal characteristics. The higher trace metal concentrations in deposits in 1997 reflected greater atmospheric pollution in the Belgrade urban area. PMID- 15893797 TI - Application of semipermeable membrane device for assessing toxicity in drinking water. AB - Semipermeable membrane devices (SPMDs) mimic passive diffusive transport of bioavailable hydrophobic organic compounds through biological membranes and their partitioning between lipids and environmental levels. Our study was developed on a surface water treatment plant based in Turin, Northern Italy. The investigated plant treats Po River surface water and it supplies about 20% of the drinking water required by Turin city (about one million inhabitants). Surface water (input) and drinking water (output) were monitored with SPMDs from October 2001 to January 2004, over a period of 30 days. The contaminant residues, monthly extracted from SPMDs by dialysis in organic solvent, were tested with the Microtox acute toxic test and with the Ames mutagenicity test. Same extracts were also analyzed with gas chromatography--mass spectrometry technique in order to characterise the organic pollutants sampled, especially Polycyclic Aromatic Hydrocarbons (PAHs). Although the PAHs mean concentration is about one hundred times lower in the output samples, the mean toxic units are similar in drinking and surface water. Our data indicate that the SPMD is a suitable tool to assess the possible toxicity in drinking water. PMID- 15893798 TI - Decolourisation of simulated reactive dyebath effluents by electrochemical oxidation assisted by UV light. AB - This study is focused on the optimisation of the electrochemical decolourisation of textile effluents containing reactive dyes with the aim of making feasible technically and economically-this method at industrial scale. Coloured waters were treated in continuous at low current density, to reduce the electrical consumption. Ti/PtO(x) electrodes were used to oxidize simulated dyebaths prepared with an azo/dichlorotriazine reactive dye (C.I. Reactive Orange 4). The decolourisation yield was dependent on the dyeing electrolyte (NaCl or Na(2)SO(4)). Dyeing effluents which contained from 0.5 to 20 gl(-1) of NaCl reached a high decolourisation yield, depending on the current density, immediately after the electrochemical process. These results were improved when the effluents were stored for several hours under solar light. After the electrochemical treatment the effluents were stored in a tank and exposed under different lighting conditions: UV light, solar light and darkness. The evolution of the decolourisation versus the time of storage was reported and kinetic constants were calculated. The time of storage was significantly reduced by the application of UV light. A dye mineralization study was also carried out on a concentrated dyebath. A TOC removal of 81% was obtained when high current density was applied for a prolonged treatment with recirculation. This treatment required a high electrical consumption. PMID- 15893799 TI - A comparison of concentration ratios for technetium and nutrient uptake by three plant species. AB - Technetium (Tc) is a non-essential element for which accumulation mechanisms in plants have recently been discussed, but only from the viewpoint of existence of anion transport proteins in plant cells. In this study, using three kinds of plants (Cucumis sativus L., Raphanus sativus L., and Brassica chinensis L.), uptake of Tc and Re (a chemical analogue of Tc) were observed. The results showed that Tc and Re uptake occurred not only with water mass flow or active nutrient uptake, but also with uptake of nutrient cations such as K+. It is suggested here that most stable chemical form under aerobic conditions, TcO4-, is used in cation transport as a substitute ions, such as Cl-. After TcO4- passes through a root surface, it moves through the xylem together with cations. Due to these uptake mechanisms, Tc is highly accumulated in plants. PMID- 15893800 TI - Adsorption of microcystins by carbon nanotubes. AB - The production of cyanobacterial toxins microcystins (MCs) by cyanobacterial bloom which may promote the growth of tumor in human liver is a growing environmental problem worldwide. In this paper, the adsorption of MC-RR and LR, which were extracted from cyanobacterial cells in Dianchi Lake in China, by carbon nanotubes (CNTs), wood-based activated carbon (ACs) and clays were investigated. Compared with ACs and clay materials of sepiolite, kaolinite and talc tested, CNTs were found to have a strong ability in the adsorption of MCs. At the concentrations of 21.5 mg l(-1) MC-RR and 9.6 mg l(-1) MC-LR in 50 mmol phosphate buffer solution (pH 7.0), the adsorption amounts of MCs by CNTs with the range of outside diameter from 2 to 10nm were 14.8 and 5.9 mg g(-1), which were about four times higher than those by other adsorbents tested. It was shown that with the decrease of CNTs outside diameters from 60 to 2 nm, the adsorption amount of MCs was apparently increased, however the size of CNTs particles formed in solution declined. This result implies that the size of CNTs tube pore that is fit for the molecular dimension of MCs plays a dominant role. Furthermore the specific surface area of CNTs was also found to be a factor in the adsorption of MCs. The results suggested that the selection of suitable size of CNTs as a kind of adsorbent is very important in the efficient eliminating MCs from drinking water in future. PMID- 15893801 TI - Exposure to the organophosphorus pesticide chlorpyrifos inhibits acetylcholinesterase activity and affects muscular integrity in Xenopus laevis larvae. AB - The effect of organophosphate pesticide chlorpyrifos (CPF) on acetylcholinesterase (AChE) activity and on skeletal muscle development in Xenopus laevis larvae was studied. To achieve our purpose embryos were exposed to 100, 250 and 3000 microg/l CPF concentrations from late blastula stage (8 h postfertilization, p.f.) to stage 47 (120 h p.f.) and the appearance of AChE activity was monitored every 24 h. Compared with control, CPF treated larvae showed a dose dependent AChE inhibition during the early stages (beginning from 24 h until 120 h p.f.) that are crucial for neuromuscular development. The amount of AChE activity that can still be measured in treated larvae at stage 47 relative to that of the control, ranged from 28% in CPFs 100 microg/l to 4% in CPFs 3000 microg/l. These low AChE activities were associated with muscular damages such as reduced myotome size and hypertrophies coupled with extensive vacuolated regions in myocytes. The occurrence of this tissue-specific injury was related to CPF concentrations and was most pronounced in CPFs 3000 microg/l which revealed a very severe AChE inhibition during the exposure. Since AChE is the major neurotransmitter of the neuromuscular system, this initial descriptive study will be an useful starting-point to ongoing and future subcellular/molecular studies that correlate the morphological damage with changes in AChE activity. PMID- 15893802 TI - XANES study of Cr sorbed by a kitchen waste compost from water. AB - A kitchen waste compost was used to sorb Cr for various times from water containing either Cr(NO3)3 or CrO3 in different concentrations. Scanning electron microscopy (SEM) results show that the composts have been partially oxidized by Cr(VI) during the sorption experiments. X-ray absorption near edge structure (XANES) simulation suggests that about 54.1-61.0% Cr sorbed on the compost is in form of organic Cr(III) through ionic exchange process with the rest being existent as Cr(NO3)3 in the Cr(III) sorption case; no Cr(OH)3 is observed or expected because the solution pH after sorption experiments is or= 5.94. Moreover, organic Cr(III) represents about 51.7-69.0% of the total sorbed Cr, and the rest (6.1-28.5%) is Cr(VI). PMID- 15893803 TI - Tissue localisation of tetra- and pentabromodiphenyl ether congeners (BDE-47, -85 and -99) in perinatal and adult C57BL mice. AB - The distribution of polybrominated diphenyl ether (PBDE) congeners was followed in C57BL mice. The animals were subjected to whole-body autoradiography using (14)C-labelled 2,2',4,4'-tetrabromodiphenyl ether (BDE-47), 2,2',3,4,4' pentabromodiphenyl ether (BDE-85) and 2,2',4,4',5-pentabromodiphenyl ether (BDE 99). Labelled BDE-85 and -99 were also used in quantitative studies on milk transfer and tissue concentrations during the neonatal period (12-15 days post partum), by use of liquid scintillation technique. The results show that in adult mice the studied PBDEs were effectively taken up, retained in fatty tissues and concentrated in some specific organs, i.e. the liver, adrenal cortex, ovary, lung and (initially) the brain. At longer post-injection time, the concentration in most tissues was considerably lower, and radioactivity was mainly found in fat depots and the liver. No significant difference in distribution between the three studied congeners was observed. Following maternal exposure, the foetal uptake was limited. On the other hand, during lactation a considerable fraction of the dose (about 20% of the studied penta-BDEs) given to the dam was transferred to the offspring. As in several cases the presently observed organ accumulation corresponds with earlier reports on PBDE effects in the same organs, the present results should be taken into consideration in the risk assessment of this compound group. PMID- 15893804 TI - Dietary restriction in rodents--delayed or retarded ageing? AB - Dietary restriction (DR) feeding increases survival significantly in strains of rats and mice. There remains however, the question as to whether these two species are always responding in an identical manner to the feeding regime. Enhanced survival can be achieved either through a set-point effect, where there is a change in the elevation of the Ln age-specific mortality rate or, by a decrease in the slope of the Ln age-specific mortality rate that results in a significant increase in the time to double the rate of mortality. It is only the second response that is evidence of a slower rate of ageing. These two possible responses to DR feeding may confound attempts to identify the biochemical mechanisms underlying the effect of DR on survival. A general lack of consistency is evident in the data and this is apparent when evaluating the free radical hypothesis of ageing in this model. Further, this hypothesis as currently viewed may be too simplistic to explain the variety and complexity of the ageing phenotype. What may be more important is not oxidative macromolecular damage but the slow transition to this cellular endpoint through the slow development of oxidative stress and the role it plays in modifying cell gene expression profiles. PMID- 15893805 TI - Food restriction, evolution and ageing. AB - The food restriction model for life extension is nearing "three-score and 10" years of age and remains in good shape, preserving much of the mystique of its youth. Although originally described for laboratory rodents, recent work shows that food restriction also appears to slow ageing processes in a range of other animal species, raising the question of whether this response represents some generalised evolutionary adaptation, perhaps a strategy to cope with periods of famine. If the food restriction response does have an adaptive basis, this would suggest that specific gene regulatory processes have evolved to shape the organism's physiological response to food restriction. It will then be important to investigate how these are organised and whether the same or different factors are at play in the various species in which food restriction extends life span. PMID- 15893806 TI - The impact of fine sediment accumulation on the survival of incubating salmon progeny: implications for sediment management. AB - This paper draws on results from a recent research programme on the impact of fine sediment transport through catchments to present a case for the development of new approaches to improving the quality of salmonid spawning and incubation habitats. To aid the development of these programmes, this paper summarises the mechanisms by which fine sediment accumulation influences the availability of oxygen (O2) to incubating salmon embryos. The results of the investigation indicate that incubation success is inhibited by: (i) the impact of fine sediment accumulation on gravel permeability and, subsequently, the rate of passage of oxygenated water through the incubation environment; (ii) reduced intragravel O2 concentrations that occur when O2 consuming material infiltrates spawning and incubation gravels; and (iii) the impact of fine particles (clay) on the exchange of O2 across the egg membrane. It is concluded that current granular measures of spawning and incubation habitat quality do not satisfactorily describe the complexity of factors influencing incubation success. Furthermore, an assessment of the trends in fine sediment infiltration indicates that only a small proportion of the total suspended sediment load infiltrates spawning and incubation gravels. This casts doubt over the ability of current catchment-based land use management strategies to adequately reduce fine sediment inputs. PMID- 15893807 TI - Excluding pulmonary embolism at the bedside with low pre-test probability and D dimer: safety and clinical utility of 4 methods to assign pre-test probability. AB - INTRODUCTION: Less than 35% of patients suspected of having pulmonary embolism (PE) actually have PE. Safe bedside methods to exclude PE could save scarce health care resources if they exclude large proportions of patients with suspected PE and are widely applicable. Non-Elisa D-dimer in combination with pre test probability of suspected PE can safely exclude PE at the bedside. Pre-test probability can be assigned by gestalt or by using clinical models (Wells, Wicki, Rodger). MATERIALS AND METHODS: We combined two databases from studies of patients with suspected PE and retrospectively compared the diagnostic test characteristics of the different methods of assigning pre-test probability. RESULTS: 535 patients were studied. PE was confirmed in 20.8% of study patients. Two clinical predictive models (Rodger and Wells) and overall diagnostic impression have similar sensitivities ranging from 96% (95% confidence interval (CI) 89-99%) to 99% (93-100%). Wicki's model has a sensitivity of 89% (77-96%). The Wells' model with a cutoff of less than 2 points in association with semi quantitative D-dimer has a specificity of 11% (CI 7-15%). The specificities for the other clinical predictive model are ranging from 21% (17-25%) to 49% (CI 42 55%). CONCLUSION: Semi-quantitative D-dimer must be combined with safe clinical probability assessment to safely exclude PE in a significant proportion of patients. Wicki's model in association with semi-quantitative D-dimer has the lowest sensitivity and should be used carefully to exclude PE at the bedside. The Wells' model with a cutoff of less than 2 points when combined with semi quantitative D-dimer excluded very few patients and therefore limits its clinical utility. PMID- 15893808 TI - ABO blood group and precocity of coronary artery disease. PMID- 15893809 TI - Histochemical localization of complex carbohydrates in the nasolabial glands of the Japanese deer (Cervus nippon yakushimae). AB - The localization of complex glycoconjugates in the nasolabial glands of the Japanese deer (C. nippon yakushimae) was studied using various histochemical methods, including lectin histochemistry, viewed using both light and electron microscopy. The secretory epithelium and luminal secretion of the deer nasolabial glands exhibited neutral and acidic glycoconjugates with different saccharide residues (alpha-D-mannose, alpha-N-acetyl-D-galactosamine, beta-D-galactose, beta N-acetyl-D-glucosamine and sialic acid). Additionally, O-acetylated sialic acids were detectable in the glandular acini. The results obtained are discussed with regard to the specific functions of the glandular secretion, which may particularly improve water retention on the skin surface and protect against physical damage as well as microbial contamination. Furthermore, our results support the view of a salivary nature of this gland type. PMID- 15893810 TI - Loss of expression of von Hippel-Lindau tumor suppressor protein associated with improved survival in patients with early-stage clear cell renal cell carcinoma. AB - OBJECTIVES: To determine whether actual expression of the von Hippel-Lindau (VHL) protein product itself (pVHL) is associated with clear cell renal cell carcinoma (CC-RCC) survival. Recent data have suggested that somatic mutations of the VHL tumor suppressor gene are associated with better cancer-specific survival in patients with CC-RCC. METHODS: Using a large, clinic-based cohort of 273 patients with CC-RCC, we tested the hypothesis that those patients with CC-RCC tumors lacking pVHL expression [pVHL(-)] will experience better cancer-specific survival than those patients with tumors that show pVHL expression [pVHL(+)]. RESULTS: Using a Cox proportional hazard model adjusting for age, patients with pVHL(-) tumors were not at a decreased risk of CC-RCC death compared with patients with pVHL(+) tumors (hazard ratio 1.0, 95% confidence interval 0.7 to 1.5). Adjustment for the Mayo SSIGN score had little effect on the risk estimate (hazard ratio 0.8; 95% confidence interval 0.5 to 1.2). In our stratified analysis, we found evidence of an inverse association with loss of pVHL expression among those patients presenting with early-stage disease (hazard ratio 0.4; 95% confidence interval 0.2 to 0.8), even after adjustment for the Mayo SSIGN score. CONCLUSIONS: Although we report no overall association, the data from this investigation are consistent with earlier findings that suggest somatic VHL alteration is associated with better cancer-specific survival among those patients presenting with early-stage (pT1 and pT2) CC-RCC. PMID- 15893811 TI - Nocturia in men with lower urinary tract symptoms is associated with both nocturnal polyuria and detrusor overactivity with positive response to ice water test. AB - OBJECTIVES: To investigate whether detrusor overactivity (DO) and the response to the ice water test (IWT) influence nighttime urinary frequency in patients with lower urinary tract symptoms. METHODS: A total of 114 patients with lower urinary tract symptoms, who were older than 50 years, with an International Prostate Symptom Score 8 or more points and a quality of life index of 2 or greater, were evaluated by a 48-hour frequency-volume chart, free flowmetry, pressure flow study, and IWT. RESULTS: The DO-positive IWT responders had a significantly greater bladder outlet obstruction index than did the DO-positive IWT nonresponders and the DO-negative IWT nonresponders. The DO-positive IWT responders had significantly more frequent nocturia and smaller nighttime maximal and minimal voided volumes than did the DO-negative IWT nonresponders without any difference in the nocturnal voided volume. The patients with nocturia two or more times had a significantly larger nocturnal voided volume and smaller nighttime minimal voided volume than the patients with nocturia less than two times. The incidence of DO-positive IWT responders was significantly greater among the patients with nocturia three or more times than that among those with nocturia less than three times. In the multivariate logistic model, the nocturnal voided volume and nighttime minimal voided volume were independently associated with nocturia two or more times and the DO-positive IWT responders were independently associated with nocturia three or more times. CONCLUSIONS: Once high-grade bladder outlet obstruction induces C-fiber-related DO, it is strongly suggested that this process, together with nocturnal polyuria, plays an important role in the consequent clinical manifestations of nocturia in patients with lower urinary tract symptoms. PMID- 15893812 TI - Therapeutic options for proximal ureter stone: extracorporeal shock wave lithotripsy versus semirigid ureterorenoscope with holmium:yttrium-aluminum garnet laser lithotripsy. AB - OBJECTIVES: To compare the safety and cost-effectiveness of ureterorenoscopic holmium:yttrium-aluminum-garnet (YAG) laser lithotripsy (URSL) with extracorporeal shock wave lithotripsy (ESWL) for proximal ureteral stones. METHODS: This investigation assessed 220 patients with upper ureteral stones. Those in the ESWL group were treated on an outpatient basis using the Medispec Econolith 2000 (Medispec, Germantown, MD) under intravenous sedation. URSL was performed with a 6/7.5F semirigid tapered ureterorenoscope and holmium:YAG laser under spinal anesthesia on an inpatient basis. A successful outcome was defined as the patient being stone free on radiography 1 month after treatment. The stone size, success rate, postoperative complications, and cost were evaluated in each group. RESULTS: A total of 220 patients were enrolled in this study. Hematuria and flank pain were the most common complaints in each group. The mean stone burden +/- SD was 58.7 +/- 3.1 mm2 in the ESWL group and 108.4 +/- 10.0 mm2 in the URSL group (P = 0.000). The accessibility of the semirigid ureterorenoscope for upper ureteral stones was 98.1% (101 of 103), and the stone-free rate achieved after one treatment was 83.2% (84 of 101). The initial stone-free rate of in situ ESWL was 63.9% (76 of 119). Significantly, the initial stone-free rate of the URSL group was superior to that of the ESWL group (P = 0.001). The average cost in the URSL group appeared to be lower than that in the ESWL group (P = 0.000). CONCLUSIONS: The results of this study have demonstrated that URSL achieved excellent results for upper ureter calculi. In terms of cost and effectiveness, this procedure should be the first-line therapy for proximal ureter stones. PMID- 15893813 TI - Polymorphism in ARE-I region of prostate-specific antigen gene associated with low serum testosterone level and high-grade prostate cancer. AB - OBJECTIVES: To examine the impact of polymorphism in the androgen-responsive element I region of the prostate-specific antigen (PSA) gene on the serum testosterone level and Gleason score in patients with newly diagnosed, untreated prostate cancer (PCa). High-grade PCa is associated with a low serum testosterone level, and the testosterone level has been negatively correlated with the expression of PSA. METHODS: Endocrine factors (including testosterone, follicle stimulating hormone, and luteotropic hormone), PSA level, prostate volume, and Gleason score were measured in 134 patients with untreated, biopsy-verified PCa. PSA polymorphism was determined by polymerase chain reaction-based methods using DNA from peripheral blood samples. RESULTS: Patients with the PSA G/G genotype had lower serum testosterone levels (3.5 +/- 1.2 ng/mL) than those with the A/A genotype (4.3 +/- 1.6 ng/mL) or the A/G genotype (4.4 +/- 1.5 ng/mL). The PSA level in the A/A and A/G genotype groups were significantly lower than that in the G/G genotype group (18.2 +/- 55.0 ng/mL versus 20.5 +/- 27.6 ng/mL, P = 0.013). In a multiple logistic regression model, the odds ratio for the G/G polymorphism was significantly increased for Gleason score (odds ratio 2.4, 95% confidence interval 1.6 to 10.4; P = 0.02) and serum testosterone level (odds ratio 0.44, 95% confidence interval 0.36 to 0.94; P = 0.01) relative to genotypes A/A and A/G. CONCLUSIONS: Our results showed that the PSA G/G genotype is associated with a greater Gleason score and serum PSA level but lower serum testosterone level and could be considered a risk factor for a poor outcome of PCa. PMID- 15893814 TI - Cocaine's effects on the perception of socially significant vocalizations in baboons. AB - The effects of cocaine on the ability of baboons to discriminate among their natural affiliative 'grunt' vocalizations were examined to determine whether cocaine would produce discrimination impairments similar to those observed previously with acoustically-similar human vowel sounds , or whether differences in cocaine's effects might occur associated with the social significance of the calls. The task employed digitized calls of actual vocalizations recorded in the wild . Baboons pressed a lever to produce a repeating 'standard' grunt, and released the lever only when one of four other 'target' grunts was selected to occur in place of the standard grunt. Cocaine (0.01-.56 mg/kg, i.m.) impaired call perception, and these impairments were more pronounced than those observed previously for acoustically-similar human vowel sounds. Cocaine also elevated reaction times as a function of dose. The results demonstrate that cocaine impairs perceptual discriminations of the natural grunt vocalizations of baboons, and suggest that cocaine may have more pronounced effects on the perception of biologically-relevant as opposed to non-relevant communication signals. PMID- 15893815 TI - The incidence of apoptosis after IVF with morphologically abnormal bovine spermatozoa. AB - Normal embryonic development depends on the maintenance of a population of normal healthy cells within each embryo. The aim of this study was to use a combination of apoptotic measures to assess differences in embryo quality after IVF with semen samples with high percentages of abnormal spermatozoa. Semen samples were obtained and cryopreserved from four Holstein bulls before (5 day prior) and after (2 week-post-insult; 2 week-PI and day 20; 3 week-PI) a scrotal insulation period of 48 h (day 0). The swim-up sperm separation method was used. The post thaw morphology revealed a decrease (P < 0.01) in the percentages of normal spermatozoa in the 3 week-PI samples in comparison with the pre-insult samples for Bulls I and III (74-22.3 and 67.7-0.5%, respectively). The percentage of vacuolated spermatozoa increased significantly for Bull II. After 18 h of sperm oocyte co-incubation, zygotes were cultured and subpopulations were removed from culture at day 8 and subjected to either the TUNEL or caspase assay. On day 8, caspase intensity increased significantly for both Bull I (217+/-147) and Bull III (229+/-98) for the 3 week-PI embryo groups compared to the equivalent embryo groups for Bull II (98+/-115) and Bull IV (90+/-111). In conclusion, the inability to consistently measure apoptosis with TUNEL alone complicated the assessment of differences in embryo quality. Thus, it is uncertain exactly when during early pre-implantation development the differences in embryo quality are first manifest. Despite discrepancies, our results clearly indicated a difference in the embryo quality between embryos obtained after IVF with semen samples from bulls that had an intense response to scrotal insulation. PMID- 15893816 TI - AC electrospray biomaterials synthesis. AB - A rapid, viable and safe fabrication method for biomaterials synthesis is reported using high-frequency AC electrospraying. We demonstrate its potential for polymeric nanoparticle fabrication, drug encapsulation in mono-dispersed micron-sized biodegradable polymer shells and the synthesis of 1microm biodegradable fibers with adjustable pore sizes as bioscaffolds for tissue/orthopaedic engineering and wound care therapy. The absence of charge in the ejected drops and fibers facilitates pulmonary drug delivery, polymer encapsulation and minimizes protein/DNA denaturing or compound ionization. PMID- 15893817 TI - Galaninergic mechanisms at the spinal level: focus on histochemical phenotyping. AB - The 29/30 amino acid neuropeptide galanin is present in a small population of DRG neurons under normal condition but is strongly upregulated after nerve injury. There is evidence that this upregulated galanin has trophic actions, for example promoting neurite outgrowth as well as influencing pain processing. In fact, both pro- and antinociceptive effects have been reported, probably relating to activation of different receptors. It has been proposed that presynaptic GalR2 receptors are pro-nociceptive by enhancing release of excitatory transmitters in the dorsal horn, and anti-nociceptive via an action on GalR1-positive interneurons. These neurons have recently been shown to be glutamatergic. Several other peptides and molecules are also regulated by nerve injury. Here we focus on neuropeptide tyrosine (NPY), which is upregulated in parallel with galanin. We review data reporting on coexistence between galanin and NPY and between these two peptides and the two NPY receptors Y1 and Y2. The data show considerable overlap, and it will be an important task to analyse how cross-talk between these neuropeptides can influence pain processing. It is proposed that such cross-talk can occur by release of peptides from DRGs neuron somata within dorsal root ganglia. To what extent these mechanisms shown to exist in rodents also occur in human is important, if one wants to discuss novel strategies for pain treatment on the basis of these findings. So far information is limited, but it has been demonstrated that galanin is expressed in DRGs and possibly also regulated. PMID- 15893818 TI - A proapoptotic function of p21 in differentiating granulocytes. AB - p21(waf 1/cip 1) (p21), best known for its ability to regulate the cell cycle, has been noted also to exert cell cycle-independent effects on apoptosis and differentiation. Inhibition of apoptosis by p21 has been reported in hematopoietic models, particularly in monocytes exposed to apoptogenic agents. The effect of p21 on survival has not hitherto been analyzed during the myeloblast to granulocyte transition. Using 32 Dc l3 murine myeloblasts, a cell line that proliferates in IL-3 and differentiates in G-CSF, we studied the effects of forced expression of p21 on cell survival. We hypothesized that exogenous p21 would suppress the modest levels of cell death associated with G CSF-mediated differentiation of 32 Dc l3 cells. Contrary to expectations, we found that exogenous p21 enhanced apoptosis of cells removed from IL-3. The p21 overexpression led to decreased cell growth, caspase-3 activation and annexin positivity. These effects occurred only in the presence of G-CSF. These findings suggest that p21 is proapoptotic in granulopoiesis, and that this effect is masked by IL-3-mediated survival signals. Our results also indicate there are distinct and opposing effects of p21 on monocytic and granulocytic survival. Aberrantly high levels of p21 may contribute to disease processes involving excessive apoptosis of granulocyte precursors. PMID- 15893819 TI - Effects of the antitumoural dequalinium on NB4 and K562 human leukemia cell lines. Mitochondrial implication in cell death. AB - Dequalinium (DQA) is a delocalized lipophylic cation that selectively targets the mitochondria of carcinoma cells. However, the underlying mechanisms of DQA action are not yet well understood. We have studied the effects of DQA on two different leukemia cell lines: NB4, derived from acute promyelocytic leukemia, and K562, derived from chronic myeloid leukemia. We found that DQA displays differential cytotoxic activity in these cell lines. In NB4 cells, a low DQA concentration (2microM) induces a mixture of apoptosis and necrosis, whereas a high DQA concentration (20microM) induces mainly necrosis. However, K562 cell death was always by necrosis as the cells showed a resistance to apoptosis at all time periods and DQA concentrations assayed. In both cell lines, the cell death seems to be mediated by alterations of mitochondrial function as evidenced by loss of mitochondrial transmembrane potential, O2*- accumulation and ATP depletion. The current study improves the knowledge on DQA as a novel anticancer agent with a potential application in human acute promyelocytic leukemia chemotherapy. PMID- 15893820 TI - Stressor controllability and learned helplessness: the roles of the dorsal raphe nucleus, serotonin, and corticotropin-releasing factor. AB - The term 'learned helplessness' refers to a constellation of behavioral changes that follow exposure to stressors that are not controllable by means of behavioral responses, but that fail to occur if the stressor is controllable. This paper discusses the nature of learned helplessness, as well as the role of the dorsal raphe nucleus, serotonin, and corticotropin-releasing hormone in mediating the behavioral effects of uncontrollable stressors. Recent research indicates that (a) uncontrollable stressors sensitize serotonergic neurons in the dorsal raphe, and that a corticotropin-releasing factor-related ligand, acting at the Type II receptor, is essential to this sensitization process, and (b) the consequent exaggerated release of serotonin in response to subsequent input is at least in part responsible for the behavioral changes that occur. Finally, implications for the general role of corticotropin-releasing hormone in stress related phenomena and for the learned helplessness paradigm as an animal model of either depression or anxiety are discussed. PMID- 15893821 TI - Psychostimulant withdrawal as an inducing condition in animal models of depression. AB - A large body of evidence indicates that the withdrawal from high doses of psychostimulant drugs in humans induces a transient syndrome, with symptoms that appear isomorphic to those of major depressive disorder. Pharmacological treatment strategies for psychostimulant withdrawal in humans have focused mainly on compounds with antidepressant properties. Animal models of psychostimulant withdrawal have been shown to demonstrate a wide range of deficits, including changes in homeostatic, affective and cognitive behaviors, as well as numerous physiological changes. Many of these behavioral and physiological sequelae parallel specific symptoms of major depressive disorder, and have been reversed by treatment with antidepressant drugs. These combined findings provide strong support for the use of psychostimulant withdrawal as an inducing condition in animal models of depression. In the current review we propound that the psychostimulant withdrawal model displays high levels of predictive and construct validity. Recent progress and limitations in the development of this model, as well as future directions for research, are evaluated and discussed. PMID- 15893822 TI - Assessing substrates underlying the behavioral effects of antidepressants using the modified rat forced swimming test. AB - Selective serotonin reuptake inhibitors (SSRIs) are the most widely prescribed antidepressant class today and exert their antidepressant-like effects by increasing synaptic concentrations of serotonin (5-HT). The rat forced swim test (FST) is the most widely used animal test predictive of antidepressant action. Procedural modifications recently introduced by our laboratory have enabled SSRI induced behavioral responses to be measured in the modified FST. The use of this model to understand the pharmacological and physiological mechanisms underlying the role of 5-HT in the behavioral effects of antidepressant drugs is reviewed. Although all antidepressants reduced behavioral immobility, those antidepressants that increase serotonergic neurotransmission predominantly increase swimming behavior whereas those that increase catacholaminergic neurotransmission increase climbing behavior. The 5-HT(1A), 5-HT(1B/1D) and 5-HT(2C) receptors are the 5-HT receptors most important to the therapeutic effects of SSRIs, based on extensive evaluation of agonists and antagonists of individual 5-HT receptor subtypes. Studies involving chronic administration have shown that the effects of antidepressants are augmented following chronic treatment. Other studies have demonstrated strain differences in the response to serotonergic compounds. Finally, a physiological model of performance in the rat FST has been proposed involving the regulation of 5-HT transmission by corticotropin releasing factor (CRF). PMID- 15893823 TI - Thinking clockwise. AB - Throughout the Animal Kingdom, the time of embryonic development is maintained and strictly controlled. Each step of the process is successful only when it occurs at the right time and place. This raises the question: how is time controlled during embryonic development? Time control is particularly crucial during embryo segmentation processes, where the number of generated segments, as well as the time of formation of each segment, is extraordinarily constant and specific for each species. Somitogenesis is the process through which the vertebrate presomitic mesoderm is segmented along its anterior-posterior axis into round-shaped masses of epithelial cells, named somites. In the chick embryo, a new pair of somites is formed every 90 min. The discovery that this clock-like precision is dictated by the somitogenesis molecular clock constituted a landmark in the Developmental Biology field. Several genes exhibit cyclic gene expression in the embryo presomitic mesoderm from which the somites arise, presenting a 90 min oscillation period, the time required to form a pair of somites. The combined levels of dynamic gene expression throughout the presomitic mesoderm enable cells to acquire positional information, thus giving them a notion of time. Anterior posterior patterning of the vertebrate nervous system also involves partition into discrete territories. This is particularly evident in the hindbrain where overt segmentation occurs. Nevertheless, little is known about the segmentation genes and mechanisms that may be involved. This paper intends to describe the molecular clock associated with vertebrate somitogenesis, suggesting that it may be operating in many other patterning processes. PMID- 15893824 TI - Gender role, sexual orientation and suicide risk. AB - BACKGROUND: There has been interest in the relationship between homosexuality, gender role and suicide risk. Though homosexuals are more likely to identify as cross-gender, research has not simultaneously examined sexual orientation and gender role in assessing suicide risk. In the current study, the unique and interactive effects of sexual orientation and gender role were assessed in regard to suicidal ideation, related psychopathology and measures of coping. METHODS: 77 participants were recruited from an undergraduate psychology subject pool (n=47) or from gay, lesbian and transgender student organizations (n=30) and assessed on measures of gender role, homosexuality, and psychopathology. RESULTS: Consistent with expectations, cross-gender role (i.e., personality traits associated with the opposite sex) is a unique predictor of suicidal symptoms. Moreover, gender role accounted for more of the overall variance in suicidal symptoms, positive problem orientation, peer acceptance and support, than sexual orientation. After accounting for gender role, sexual orientation contributed little to the variance in suicidal symptoms, associated pathology and problem-solving deficits. There was no support for gender role by sexual orientation interaction effects. LIMITATIONS: The cross-sectional nature of the data limits statements regarding causality. CONCLUSIONS: Cross-gendered individuals, regardless of sexual orientation, appear to have higher risk for suicidal symptoms. Researchers and clinicians should assess gender role in evaluations of youth samples. PMID- 15893825 TI - Obsessive-compulsive disorder symptom dimensions show specific relationships to psychiatric comorbidity. AB - The goals of this study were to examine relationships among symptom categories in obsessive-compulsive disorder (OCD), to establish OCD symptom dimensions by factor- and cluster-analytic analyses, and to explore associations between OCD symptom dimensions and comorbid neuropsychiatric conditions. A total of 317 OCD participants underwent a systematic diagnostic interview using the Structured Clinical Interview for DSM-IV. OCD symptoms assessed by the Yale-Brown Obsessive Compulsive Scale Symptom Checklist (N=169) and by the Thoughts and Behaviors Inventory (N=275) were subjected to factor and cluster analyses. An identical four-factor solution emerged in two different data sets from overlapping samples, in agreement with most smaller factor-analytic studies employing the YBOCS checklist alone. The cluster analysis confirmed the four-factor solution and provided additional information on the similarity among OCD symptom categories at five different levels. OCD symptom dimensions showed specific relationships to comorbid psychiatric disorders: Factor I (aggressive, sexual, religious and somatic obsessions, and checking compulsions) was broadly associated with comorbid anxiety disorders and depression; Factor II (obsessions of symmetry, and repeating, counting and ordering/arranging compulsions) with bipolar disorders and panic disorder/agoraphobia; and Factor III (contamination obsessions and cleaning compulsions) with eating disorders. Factors I and II were associated with early onset OCD. This study encourages the use of cluster analyses as a supplementary method to factor analyses to establish psychiatric symptom dimensions. The frequent co-occurrence of OCD with other psychiatric disorders and the relatively specific association patterns between OCD symptom dimensions and comorbid disorders support the importance of OCD subtyping for treatment, genetic, and other research studies of this heterogeneous disorder. PMID- 15893826 TI - Production of a monoclonal antibody against SARS-CoV spike protein with single intrasplenic immunization of plasmid DNA. AB - Severe acute respiratory syndrome (SARS) is a highly infectious disease caused by a novel coronavirus (SARS-CoV). Specific monoclonal antibodies (mAbs) against the SARS-CoV are vital for early diagnosis and pathological studies of SARS. Direct intrasplenic inoculation of plasmid DNA encoding antigen is an effective and fast approach to generate specific mAb when the protein antigen is difficult to prepare or dangerous in use. In this study, we selected one fragment of SARS-CoV spike protein (S1-(3)) as antigenic determinant by immunoinformatics. Single intrasplenic immunization of plasmid DNA encoding S1-(3) induced anti-spike protein antibodies. We established one hybridoma cell line secreting specific mAb and evaluated this mAb with murine leukemia virus pseudotyped with SARS-CoV spike protein (MLV/SARS-CoV). The mAb could recognize the spike protein on the MLV/SARS CoV-infected Vero E6 cells albeit with no neutralizing effect on the infectivity of the pseudotype virus. Our results show that a single-shot intrasplenic DNA immunization is efficient for the production of specific mAb against SARS spike protein, and a linear epitope of the spike protein is recognized in this study. PMID- 15893827 TI - Implicit versus explicit ranking: on inferring ordinal preferences for health care programmes based on differences in willingness-to-pay. AB - The paper explores the merit of the willingness-to-pay (WTP) method as a way to elicit public preferences regarding health care priorities. The aim is to test the extent to which the implicit ranking inferred from the ordinal differences in WTP-values corresponds with respondents' explicit ranking of the same programmes. This issue of convergent validity is explored by face-to-face interviewing of population samples in six European countries-in total 1240 respondents. The most consistent result is the inconsistency of WTP and explicit ranking in all six countries. The convergent validity of WTP is low, particularly among those who did not state different WTP-values on the three programmes being considered. PMID- 15893828 TI - An extremely rapid and simple DNA-release method for detection of M. tuberculosis from clinical specimens. AB - A single-step, 5-min lysis method was investigated as a rapid technique to extract genomic DNA from mycobacteria for PCR detection of M. tuberculosis directly from clinical specimens. Of 67 smear-positive clinical specimens, 64 (95.5%) were positive by PCR after this rapid extraction method. PMID- 15893829 TI - Fabrication and optimization of the multiplex PCR-based oligonucleotide microarray for detection of Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum. AB - To detect and identify the pathogens responsible for sexually transmitted diseases (STDs) at the early stage of infection and with a high throughput, a new microarray with a bifunctional probe modification was prepared using Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum as a model system. During the fabrication of the microarray, an asymmetric fluorescently labeled multiplex PCR was introduced. The fabrication optimization proved that the best hybridization results would be obtained by spotting N. gonorrhoeae probe at a position near the side of the fluorescently labeled reverse primer within its target gene and spotting each probe at a concentration of 50 microM onto the aldehyde-derived glass slides using spotting solution S1 and using hybridization solution H2 for hybridization. The probes designed by our laboratory could specifically discriminate the pathogens of N. gonorrhoeae, C. trachomatis and U. urealyticum in the presence of the internal control on the microarray simultaneously and separately. By incorporating the key features of DNA microarray with those of multiplex PCR, the microarray provides a fast high throughput platform for multiple infections and multiple samples to be detected and identified simultaneously for STD clinics. It also provides a new platform for other diseases and gene mutations to be detected and identified at a high throughput. PMID- 15893830 TI - PCR-DGGE analysis for the identification of microbial populations from Argentinean dry fermented sausages. AB - Different PCR-DGGE protocols were evaluated to monitor fermentation process and to investigate bacterial communities developed in two artisanal Argentinean fermented sausages. Bacterial universal primers frequently used in PCR-denaturing gradient gel electrophoresis (DGGE) were evaluated. Lactic acid bacteria (LAB) and staphylococci species isolated from Tucuman sausages were used to determine the experimental conditions for PCR amplification and DGGE differentiation. Total microbial DNA extracted directly from both fermented sausages was subjected to DGGE analysis. PCR-DGGE results were different for each set of primers used. Primers Bact-0124f(GC)-Uni-0515r and V1f(GC)-V1r showed to be efficient to differentiate LAB and Staphylococcus cultures while the set V3f(GC)-Uni-0515r allowed to demonstrate the succession of different Lactobacillus and Staphylococcus species during ripening process. An intense band corresponding to Lactobacillus sakei was observed to be present in both samples. Staphylococcus saprophyticus was only observed in Tucuman sausage while a band identified as Brochothrix thermophacta was detected in Cordoba sausage. PCR-DGGE analysis of different 16S rDNA amplicons was able to discriminate between LAB and Gram positive, coagulase-negative cocci, resulting an effective tool to establish the microbiota developed in artisanal dry sausages. PMID- 15893831 TI - Real-time detection of common microbial volatile organic compounds from medically important fungi by Selected Ion Flow Tube-Mass Spectrometry (SIFT-MS). AB - We describe a new method, Selected Ion Flow Tube-Mass Spectrometry (SIFT-MS) for the rapid and sensitive real-time detection and quantification of volatile organic compounds from medically important fungi, grown on a range of laboratory media. SIFT-MS utilises the chemical ionisation reactions of mass-selected ions to characterise volatile organic compounds (VOCs) that are produced as metabolites from fungi. This technique has the distinct advantage over others in that it readily detects low molecular weight, reactive volatiles, and allows for real-time, quantitative monitoring. The fungi examined in this study were Aspergillus flavus, Aspergillus fumigatus, Candida albicans, Mucor racemosus, Fusarium solani, and Cryptococcus neoformans grown on or in malt extract agar, Columbia agar, Sabouraud's dextrose agar, blood agar, and brain-heart infusion broth. Common metabolites (ethanol, methanol, acetone, acetaldehyde, methanethiol, and crotonaldehyde) were detected and quantified. We found the fingerprint of volatiles, in terms of presence and quantity of volatiles to be strongly dependent on the culture medium, both in terms of variety and quantity of volatiles produced, but may form the basis for species specific identification of medically important fungi. PMID- 15893832 TI - Overview of national guidelines for infrastructure and staffing of radiotherapy. ESTRO-QUARTS: work package 1. AB - BACKGROUND AND PURPOSE: The aim of this study was to collect the available guidelines for infrastructure and staffing throughout Europe and to develop general guidelines. MATERIALS AND METHODS: Questionnaires were sent to representatives in all European countries. Countries were divided into three groups, based on the GDP per capita (low, medium and high resource countries). Based on the results, general guidelines were developed. RESULTS: 41 countries (93.2%), representing over 99% of the European population responded. Guidelines for the number of linear accelerators and personnel were available in about 40% of the countries. For accelerators, the average was 1 per 183,000 inhabitants in the high, 1 per 284,000 in the medium and 1 per 500,000 in the low resource countries. For radiation oncologists, it varied between 1 per 150-400 patients (average: 250) and there was no clear difference between high, medium and low resource countries. For physicists, the average was 1 per 530 patients for high and 1 per 610 for medium or low resource countries. Guidelines for other equipment and staffing are highly dependent on local habits and complexity of the treatments. CONCLUSION: An accurate overview of existing guidelines for radiotherapy infrastructure and staffing is obtained. It is suggested to have one linear accelerator per 450 patients, one radiation oncologist per 200-250 patients and one physicist per 450-500 patients (or one per linac). It should be stressed, however, that these are only crude guidelines and that the actual needs heavily depend on population structure, cancer incidence and treatment strategies, which differ between the various countries. PMID- 15893833 TI - Treatment outcome after linac-based radiosurgery in cerebral arteriovenous malformations: retrospective analysis of factors affecting obliteration. AB - PURPOSE: We investigated patient outcome and factors affecting obliteration rate after radiosurgery in cerebral arteriovenous malformations (AVM). MATERIAL AND METHODS: We have treated 110 patients with cerebral AVM with linac-based radiosurgery (RS). AVM classification according Spetzler-Martin was 17 patients grade I (15%), 39 grade II (36%), 41 grade III (37%), 12 grade IV (11%) and 1 grade V (1%). Median single dose was 18 Gy. Mean treatment volume was 4.7 cc (range, 0.1-24.0 cc). Fifty-two patients experienced hemorrhage prior to RS. Median follow-up was 2.5 years. RESULTS: Actuarial complete obliteration rate (CO) was 51% after 3 years and 67% after 4 years. CO rate was significantly higher in AVM <3 cm (64% vs. 43%, P<0.04) and in patients with grade I/II vs. III V (71% vs. 33%, P<0.001). CO was significantly improved after doses >18 Gy (P<0.02) and in male gender (P<0.04). In multivariate analysis Spetzler-Martin grade remained significant. Intracranial hemorrhage after RS occurred in 9 patients 13.9 months median after RS. Neurological dysfunction improved/completely dissolved or remained stable in 95% of patients. No new onset of neurological dysfunction was seen. No significant adverse effects after RS were seen. CONCLUSIONS: The rate of obliteration after RS in AVM depends on applied single dose as well as size and Spetzler-Martin grade. RS is an alternative to neurosurgery, especially in patients with small or surgically inaccesible AVM. PMID- 15893834 TI - Binocular rivalry between emotional and neutral stimuli: a validation using fear conditioning and EEG. AB - When two incompatible pictures are projected to the two eyes, they compete for perceptual dominance. Previous research has claimed that meaningful and emotionally valenced pictures predominate over neutral pictures in this rivalry. This may be interpreted as evidence for preferential processing of emotionally significant stimuli in the visual system but it is difficult to dismiss that the physical characteristics of the different pictures or response biases influenced the results of these studies. Thus, we set out to examine the influence of emotion using methods eliminating the influence of physical characteristics and minimizing response biases. We used simple visual patterns and induced emotional valence by fear conditioning. In Experiment 1 the aversive CS+ predominated over the CS-. In Experiment 2 we extended previous findings by showing that participants' self-reported perception is validated by corresponding steady-state visually evoked potentials in the EEG in the context of such a conditioning experiment. This was accomplished by frequency coding the rivalling stimuli with a stimulus specific pattern reversal and extracting the corresponding frequency from the occipital lobe EEG. Taken together, these studies provide further evidence that picture valence can influence perception in binocular rivalry. This is discussed in terms of subcortical mechanisms supporting the efficient processing of threatening information. PMID- 15893835 TI - Quantitative analysis of functional clustering of neurons in the macaque inferior temporal cortex. AB - Neurons with similar preferences for two-dimensional shapes of intermediate complexity cluster in area TE of the monkey inferior temporal cortex. To further characterize the functional structure of area TE, we quantitatively analyzed various aspects of the visual responses of closely located neurons by applying multiple single-unit recording techniques in anesthetized monkeys. Examination of the visual responses elicited with a large, predetermined set of visual stimuli confirmed previous findings that nearby neurons, on average, exhibited positively correlated preferences for a set of visual stimuli. Nearby neurons also tended to be similar in their receptive-field organization and contrast-polarity preference. In contrast, no correlation was found in the size tuning of neighboring neurons. Pooling or subtraction of activities between a pair of nearby neurons was shown to improve stimulus discriminability, if the neuron pair had positively or negatively correlated stimulus preferences, respectively. These results indicate that nearby TE neurons share some aspects of stimulus preference, but their response selectivity differ in other aspects. Both pooling and subtraction between nearby neurons can reduce across-trial response variability, if these decoding strategies are applied to appropriate neuronal pools. PMID- 15893836 TI - Topological relation of chick thalamofugal visual projections with hyper pallium revealed by three color tracers. AB - In birds, there are two visual projections from retina to higher pallium, i.e., tectofugal and thalamofugal pathways. The latter one is lateralized in chick and suggested to be involved in visually evoked social behavior, like recognition of novelty, predator, and conspecific animals. We wanted to establish functionally relevant topological connection map between thalamic nuclei and hyperpallium apicale (HA) and carried out tracing study with three color fluorescent tracers. The tracers were serially injected in HA either along with the medial-lateral (M L) or anterior-posterior (A-P) axis. We found that M-L axis and A-P axis in HA were transferred into the dorsal-ventral axis and the medial-lateral axis, respectively within the nucleus geniculatus lateralis pars dorsalis (GLd). In another word, the medial part of nucleus dorsolateralis anterior thalami pars lateralis (DLLv) projected to anterior part of HA and the ventral part of nucleus dorsolateralis anterior thalami pars lateralis pas ventralis (DLLd) projects to lateral HA. This result suggests that thalamus would process information in parallel through each subnuclei and elaborate coordination among them in relation to topological map presented in higher pallium. PMID- 15893837 TI - Molecular mechanisms of measles virus persistence. AB - As measles virus causes subacute sclerosing panencephalitis and measles inclusion body encephalitis due to its ability to establish human persistent infection, without symptoms for the time between the acute infection and the onset of clinical symptoms, it has been the paradigm for a long term persistent as opposed to chronic infection by an RNA virus. We have reviewed the mechanisms of persistence of the virus and discuss specific mutations associated with CNS infection affecting the matrix and fusion protein genes. These are placed in the context of our current understanding of the viral replication cycle. We also consider the proposed mechanisms of persistence of the virus in replicating cell cultures and conclude that no general mechanistic model can be derived from our current state of knowledge. Finally, we indicate how reverse genetics approaches and the use of mouse models with specific knock-out and knock-in modifications can further our understanding of measles virus persistence. PMID- 15893838 TI - Theiler's virus persistence in the central nervous system of mice is associated with continuous viral replication and a difference in outcome of infection of infiltrating macrophages versus oligodendrocytes. AB - Theiler's murine encephalomyelitis virus (TMEV) infection of mice, in which persistent central nervous system (CNS) infection induces Th1 CD4+ T cell responses to both virus and myelin proteins, provides a relevant experimental animal model for MS. During persistence, >10(9) TMEV genome equivalents per spinal cord are detectable by real-time reverse transcription-polymerase chain reaction (RT-PCR). Because of the short half-life of TMEV (<1 day), continual viral replication is needed to sustain these very high TMEV copy numbers. An essential role for macrophages in TMEV persistence has been documented and, although limited by host anti-viral immune responses, TMEV nonetheless spreads during persistence to infect other cells, particularly oligodendrocytes, in which the infection is productive and lytic. Virus factors influencing persistence of TMEV are expression of the out-of-frame L* protein and use of sialic acid co receptors. PMID- 15893839 TI - Transport of nerve growth factor encapsulated into liposomes across the blood brain barrier: in vitro and in vivo studies. AB - A nerve growth factor (NGF) was encapsulated into liposomes in order to protect it from the enzyme degradation in vivo and promote it permeability across the blood-brain barrier (BBB). RMP-7, a ligand to the B2 receptor on brain microvascular endothelial cells (BMVEC), was combined with 1,2-dioleoyl-sn glycero-3-phosphoethanolamine-n-[poly(ethylenegly-col)]-hydroxy succinamide (DSPE PEG-NHS) to obtain DSPE-PEG-RMP-7. Then DSPE-PEG-RMP-7 was incorporated into the liposomes' surface to target sterically stabilized liposomes (SSL-T) to the brain. The highest percent of NGF encapsulated into liposomes was about 34%, and the average size of liposomes was below 100 nm. A primary model of BBB was established and evaluated by morphological, permeability, and transendothelial electrical resistance (TEER). The BBB model was employed to study the permeability of NGF liposomes in vitro. The results indicated that the liposomes could enhance transport of NGF across the BBB. The best transport rate was received with NGF-SSL-T. The brain distribution of NGF liposomes was studied in vivo, the amount of NGF in the brain was increased in the order: NGF-SSL-T>NGF SSL+RMP-7>NGF-SSL>NGF-L. The maximum concentration of NGF was recorded in 30 min following the intravenous injection. In particular, a majority of NGF was distributed in striatum, hippocampus and cortex, and the concentration of NGF was relatively lower in olfactory bulb, cerebellum and brain stem. There was a close relationship between P(e) (permeability coefficient on in vitro BBB model) and T(e) (brain targeted coefficient in vivo) for NGF encapsulated into the liposomes. PMID- 15893840 TI - Heparin-loaded zein microsphere film and hemocompatibility. AB - Zein was studied as a drug-eluting coating film composed of zein microspheres for cardiovascular devices (e.g. stent). In vitro 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) analysis showed that both zein film and its degraded product had better biocompatibility compared with Corning culture plate on the growth of human umbilical veins endothelial cells (HUVECs, p<0.05, n=6), and the effect of zein degraded product on HUVECs was dose-dependent. The best result was obtained at 0.3 mg/ml of the addition. The encapsulation efficiency of heparin and heparin loading varied with the amount of both zein and heparin, and the highest encapsulation efficiency (heparin 1.33 mg/ml and zein 16 mg/ml) was 22.77+/-1.33% (n=3). Scanning electron microscope (SEM) observation indicated that the zein film was made of microspheres in diameter from nano- to micrometer, which could be controlled. Sizes of heparin-loaded zein microspheres changed before and after release of heparin because of conglutination among zein microspheres. Release rate of heparin from microsphere film reached to 33.5+/ 1.2% within 12 h, and began to get into subsequent "slow release" phase; about 55% of the entrapped heparin was released after 20 days. Both zein film and heparin-loaded zein microsphere film were effective in suppressing platelet adhesion, and the heparin-loaded film showed a better anticoagulation as determined with thrombin time (TT) assay. These results suggest that zein film could be used directly as a new type of coating material for its better biocompatibility with HUVECs. Moreover, the heparin-loaded zein microsphere film can significantly improve the hemocompatibility. PMID- 15893841 TI - Inhibition of VEGF receptor-2 decreases the development of hyperdynamic splanchnic circulation and portal-systemic collateral vessels in portal hypertensive rats. AB - BACKGROUND/AIMS: Portal hypertension is characterized by the development of a hyperdynamic splanchnic circulation. To determine whether this process is angiogenesis-dependent, we assessed the effects of SU5416, a specific inhibitor of VEGF receptor-2, in portal hypertensive rats. METHODS: Rats with portal hypertension induced by partial portal vein ligation were treated with SU5416 or vehicle during 5 days. Then, hemodynamic studies were performed using radioactive microspheres. Protein expressions of CD31, VEGF receptor-2 and VEGF were also determined by Western blotting. RESULTS: Treatment of portal hypertensive rats with SU5416 resulted in a significant and marked decrease (by 44%) in portal venous inflow, and increases in splanchnic arteriolar resistance (by 68%) and portal venous resistance (by 93%). In addition, SU5416 administration significantly inhibited the formation of portal-systemic collateral vessels (52% inhibition), as well as the splanchnic CD31 and VEGF receptor-2 protein expressions in portal hypertensive rats, compared with those receiving vehicle. CONCLUSIONS: This study demonstrates that the development of hyperdynamic splanchnic circulation and the formation of portal-systemic collateral vessels in portal hypertensive rats are angiogenesis-dependent processes that can be markedly inhibited by blockade of the VEGF signaling pathway. Therefore, modulation of angiogenesis may represent a potential target in the treatment of portal hypertension. PMID- 15893842 TI - Taurine inhibits oxidative damage and prevents fibrosis in carbon tetrachloride induced hepatic fibrosis. AB - BACKGROUND/AIMS: The aim of the study was to examine the effects of taurine on hepatic fibrogenesis and in isolated hepatic stellate cells (HSC). METHODS: The rats of the hepatic damage (HD) group were administered carbon tetracholoride (CCl4) for 5 weeks and a subgroup received, in addition, a 2% taurine containing diet for 6 weeks (HDT). The HSC were isolated from normal rats and cultured for 4 days. RESULTS: The hepatic taurine concentration was decreased in the HD group. This loss and the hepatic histological damage and fibrosis (particularly in the pericentral region), were reduced following taurine treatment. Furthermore, the hepatic alpha-SMA, lipid hydroperoxide and 8-OHdG levels in serum and liver, as well as hepatic TGF-beta1 mRNA and hydroxyproline levels were significantly increased in the HD group, and most of these parameters were significantly reduced following taurine treatment. In contrast to the MAP-kinase and Akt expressions, which remained unchanged, the lipid hydroperoxide and hydroxyproline concentrations, as well as TGF-beta1 mRNA levels were significantly reduced by taurine in activated HSC. CONCLUSIONS: Oral taurine administration enhances hepatic taurine accumulation, reduces oxidative stress and prevents progression of hepatic fibrosis in CCl4-induced HD rats, as well as inhibits transformation of the HSC. PMID- 15893843 TI - VEGF-induced angiogenesis drives collateral circulation in portal hypertension. PMID- 15893844 TI - A novel sodium overload test predicting ascites decompensation in rats with CCl4 induced cirrhosis. AB - BACKGROUND/AIMS: We aimed to develop a non-invasive test to identify the initial alterations of sodium homeostasis and prospectively predict decompensation in preascitic cirrhotic rats. METHODS: The sodium overload test (SOT) was performed in control (CT) and CCl4-induced cirrhotic rats (CH) by calculating the percentage of sodium excreted in the urine after NaCl oral administration (0.5 g/kg). Liver fibrosis was quantified by image cytometry. RESULTS: From the 8th week of CCl4 intoxication, while the daily sodium balance did not change in CH and CT, SOT became significantly lower in the former (62.1+/-13.2 vs 78.8+/ 13.2%; P=0.035). At sacrifice, ascites was only present in one animal. The degree of liver fibrosis correlated with SOT. In subsequent experiments, 17 cirrhotic rats developed ascites between the 9th and 14th weeks. SOT remained stable up to 3 weeks before ascites appearance, while it fell significantly to 35+/-19 and 26+/-21% at 2 and 1 week before ascites diagnosis, respectively. Nearly all the rats (95%) with a SOT<60% developed ascites within 3 weeks. CONCLUSIONS: In preascitic cirrhotic rats, SOT unveils sodium metabolism abnormalities earlier than the daily sodium balance and prospectively predicts ascites appearance, identifying rats in a homogeneous stage of cirrhosis, which is essential in pathophysiological studies on sodium retention. PMID- 15893845 TI - Morpholino oligonucleotide-triggered beta-catenin knockdown compromises normal liver regeneration. AB - BACKGROUND/AIMS: Wnt/beta-catenin activation is seen during early liver regeneration (LR) observed as stabilization and translocation to the nucleus followed by an overall decrease. However, beta-catenin continues to be in hepatocyte nucleus and membrane, secondary to its increased gene expression at 6 72 h. METHODS: In the present study, we examined the effect of ablating beta catenin transcription on LR. Twelve male fisher rats were subjected to two-third partial hepatectomy followed by administration of beta-catenin antisense phospho morpholino oligonucleotide (AS) in six or mismatch control (CON) injection in the remaining 6 via superior mesenteric vein. Three animals from each group were sacrificed at 24 h and 7 days for liver assessment. RESULTS: AS group exhibited a significant decrease in total beta-catenin at 24 h. A significant decrease in liver/body weight ratio was also observed in the AS group at 24 h and 7 days that was due to decreased proliferation. Among the targets of this pathway c-myc and uPAR levels showed significant decrease while cyclin-D1 remained unaffected. CONCLUSIONS: We demonstrate the importance of beta-catenin in early liver regeneration especially in hepatocyte proliferation. Also, c-myc and uPAR might be crucial downstream effectors of beta-catenin during liver regeneration. PMID- 15893846 TI - Treatment of acute C hepatitis in intravenous drug users. PMID- 15893847 TI - Differentiation of rat bone marrow cells cultured on artificial basement membrane containing extracellular matrix into a liver cell lineage. AB - BACKGROUND/AIMS: Bone marrow (BM) cells have been shown to be capable of differentiating into a liver cell lineage in vitro. However, their differentiation and proliferation is poor, and the cell characteristics are poorly understood. METHODS: We cultured rat BM cells on an artificial basement membrane containing extracellular matrix (ECM) with hepatocyte growth factor (HGF). The expression of mRNA for liver-specific genes was analyzed by reverse transcription PCR. The expression of albumin and Musashi-1 by cultured cells was analyzed using a fluorescence-activated cell sorter (FACS). The proportions of albumin-positive cells when culture was performed with different concentrations of HGF were analyzed by FACS. RESULTS: On culture day 21, polygonal cells proliferated and formed cell colonies. These cells expressed mRNA for all the liver-specific genes analyzed, and showed heterogeneous differentiation, some cells expressing albumin, others expressing Musashi-1. Albumin-positive differentiated cells were large and rich in intracellular structures, while Musashi-1-positive undifferentiated cells had the opposite characteristics. Culturing cells with higher concentrations of HGF induced an increased proportion of albumin-positive cells. CONCLUSIONS: The results suggest that cell culture on an ECM with a high concentration of HGF increases the extent to which BM cells differentiate into a liver cell lineage and proliferate in vitro. PMID- 15893848 TI - Life expectancy and health care expenditures: a new calculation for Germany using the costs of dying. AB - Some people believe that the impact of population ageing on future health care expenditures will be quite moderate due to the high costs of dying. If not age per se but proximity to death determines the bulk of expenditures, a shift in the mortality risk to higher ages will not affect lifetime health care expenditures as death occurs only once in every life. We attempt to take this effect into account when we calculate the demographic impact on health care expenditures in Germany. From a Swiss data set, we derive age-expenditure profiles for both genders, separately for persons in their last 4 years of life and for survivors, which we apply to the projections of the age structure and mortality rates for the German population between 2002 and 2050 as published by the Statistische Bundesamt. In the extreme case, we assume that morbidity is compressed at the end of life in such a way that a 60-year old in 2050 is as healthy as a 56-year old today if his life expectancy is 4 years higher. We calculate that at constant prices, per-capita health expenditures of Social Health Insurance would rise from 2596 Euro in 2002 to between 2959 Euro and 3102 Euro in 2050 when only the age structure of the population changes and everything else remains constant at the present level, and to between 5232 Euro and 5485 Euro with a technology-driven exogenous cost increase of 1% per annum. A "naive" projection based only on the age distribution of health care expenditures, but not distinguishing between survivors and decedents, yields values of 3217 Euro and 5688 Euro for 2050, respectively. Thus, the error of excluding the "costs of dying" effect is small compared with the error of underestimating the financial consequences of expanding medical technology. PMID- 15893849 TI - A conserved mRNA expression profile of SREB2 (GPR85) in adult human, monkey, and rat forebrain. AB - SREB is a subfamily of G-protein-coupled receptors, which consists of SREB1 (GPR27), SREB2 (GPR85), and SREB3 (GPR173). Its high evolutionary conservation and predominant expression in the CNS suggest that SREB family members and their undiscovered ligand(s) may have significant functions in the nervous system. SREB2 is the most conserved receptor throughout vertebrate evolution. As a first step in understanding the function of the SREB family, we have determined the anatomical gene expression profile of SREB2 in adult human, monkey, and rat forebrain using in situ hybridization histochemistry. The expression pattern of SREB2 mRNA was well conserved across three mammalian species. SREB2 mRNA was expressed in neurons throughout the brain and the most abundant expression was detected in the hippocampal dentate gyrus in all species examined. The areas expressing high levels of SREB2 mRNA overlap with brain structures known to possess high levels of plasticity, namely, the hippocampal formation, olfactory system, and supraoptic and paraventricular nuclei. Further, the anatomical expression of SREB1 and SREB3 overlapped with that of SREB2 in the adult monkey brain. Together, these data suggest a possible link between SREB family and neural plasticity, which may explain its extremely high conservation throughout vertebrate evolution. PMID- 15893850 TI - Outbreaks of contaminated broncho-alveolar lavage related to intrinsically defective bronchoscopes. AB - From 5 March 2001 to 19 October 2001, outbreaks of broncho-alveolar lavage (BAL) contamination with Enterobacteraceae were detected in our 700-bed institution. We report the investigation of these outbreaks. A case was defined as the occurrence of pairs of specific Enterobacteraceae in BAL specimens among any patients who underwent bronchoscopy in the respiratory unit during the period of the outbreak. Contamination was identified in 117 BAL samples during three outbreaks among 418 patients, and was associated with bronchoscopes 11 and 12 (P<0.001). The other five devices in use were not linked with the outbreaks. During the first outbreak, particular pairs of micro-organisms were associated with a specific bronchoscope (Klebsiella pneumoniae/Proteus vulgaris with bronchoscope 11, and Morganella morganii/Proteus mirabilis with bronchoscope 12). Cultures of sputa from two patients also yielded M. morganii some days after bronchoscopic examination. Isolates from contaminated BAL samples and bronchoscope 11 had similar patterns by pulsed-field gel electrophoresis. No further cases occurred after removal of the implicated bronchoscopes. No deficiencies in disinfection procedures were detected and the source of contamination was found to be a loose port of the biopsy channel of the bronchoscope. Our findings underscore the urgent need to test bronchoscopic samples regularly and to improve the design and structure of bronchoscopes. PMID- 15893851 TI - Acinetobacter baumannii: emergence and spread in Israeli hospitals 1997-2002. AB - The incidence of multi-drug-resistant Acinetobacter baumannii bloodstream infections (BSIs) increased two- to four-fold in three Israeli hospitals between 1997 and 2002, accounting for 3.5-18% of all hospital-acquired BSIs. This was associated with increasing carbapenem resistance reaching 35-54%, and by a dramatic increase in carbapenem consumption. In-hospital fatality rates ranged between 47% and 58% and were significantly higher than those seen with other nosocomial Gram-negative pathogens. A. baumannii was not restricted to intensive care units, but had spread to all hospital wards. Multi-drug-resistant A. baumannii has the potential to reach endemicity in hospitals and warrants more vigorous and innovative efforts to limit its spread. PMID- 15893852 TI - Colonization by high-level aminoglycoside-resistant enterococci in intensive care unit patients: epidemiology and clinical relevance. AB - A cohort study was performed to investigate the risk factors for colonization with high-level aminoglycoside-resistant enterococci (HLARE) in intensive care unit (ICU) patients. Colonization was investigated by performing surveillance samples during ICU stay. Clonal relatedness of the isolates was assessed by pulsed-field gel electrophoresis. Eighty-six patients with an ICU stay of >48 h were included; two were colonized with HLARE at admission, and 24 (28.5%) acquired HLARE during their stay in the ICU. HLARE were initially isolated from rectal swabs alone. Thirty-five percent of Enterococcus faecalis and 57% of E. faecium showed high-level resistance to gentamicin or streptomycin. Most isolates were clonally unrelated. Using multi-variate analysis, the only variable associated with HLARE colonization was previous antimicrobial use. Five patients had HLARE isolated from clinical samples, three of them with infection; in all of these, colonization with the same clone had been detected previously by surveillance samples. We conclude that most infections due to HLARE in the ICU are preceded by previous colonization, and that antimicrobial use is the main risk factor for colonization. PMID- 15893853 TI - Emergence of antibiotic resistance amongst hospital-acquired urinary tract infections and pharmacokinetic/pharmacodynamic considerations. AB - Bacterial urinary tract infections (UTIs) are frequent infections in the nosocomial setting. Nosocomial UTIs are almost exclusively complicated UTIs, although the complicating factors may be very heterogenous. The bacterial spectrum of nosocomial UTIs is broad and antibiotic resistance is common. The results of international and national surveillance studies on the bacterial spectrum and antibiotic resistance of nosocomial uropathogens are provided. The treatment of nosocomial UTIs encompasses treatment of the complicating factors as well as antimicrobial chemotherapy. At least in serious UTIs, adequate initial antibiotic therapy results in lower mortality. Therefore, the initial antibiotic regimen must provide sufficient antibiotic cover. However, this can only be achieved if the bacterial spectrum and antibiotic resistance patterns of uropathogens in the institution are followed continuously. Provisional microbiological findings, such as reports on Gram stain or certain biochemical results, can lead to early stratification of pathogens and allow more tailored empiric antibiotic therapy. Antibiotic therapy of nosocomial UTIs has to consider two different aspects: (1) therapeutic success in the individual patient; and (2) prevention of emergence of antibiotic-resistant mutants. The emergence of resistance can possibly be lowered by adequate drug selection and dosing. Increasing antibiotic resistance requires more prudent use of antimicrobial drugs. PMID- 15893854 TI - Competition between methicillin-sensitive and -resistant Staphylococcus aureus in the anterior nares. AB - Colonization of the anterior nares with Staphylococcus aureus has been shown to be a risk factor for infection. The purpose of this study was to test the hypothesis that methicillin-resistant strains of S. aureus (MRSA) compete with methicillin-sensitive (MSSA) strains for colonization of the anterior nares. As part of the local National Health Service trust MRSA infection control strategy, patients who have been in a healthcare institution in the last year are routinely sampled and tested for MRSA colonization at the time of hospital admission. The sampling and testing methods were modified for the six-month period of this study to allow the detection of both MSSA and MRSA/MSSA co-colonization. MRSA alone was carried by 56 (8%) of 680 patients, MSSA alone by 115 patients (17%), 505 patients (74.3%) carried neither, and four patients (0.6%) carried both MRSA and MSSA. The deviance between the observed number of co-colonized swabs and that expected under the null hypothesis of no competition between MSSA and MRSA was significant (P=0.02, Fisher's exact test). The statistical approach is unaffected by the confounding effect of factors that affect the relative frequencies of MRSA or MSSA colonization. When logistic regression was used to estimate the extent of competition, controlling for effects of age and sex, we estimated a protective efficacy of MSSA colonization in the prevention of MRSA colonization of 78% (95% CI 29-99%). Results from this cross-sectional study support the hypothesis that MRSA and MSSA compete for colonization space, and provides an estimate of the extent to which MSSA interferes with MRSA colonization. PMID- 15893855 TI - PVN galanin increases fat storage and promotes obesity by causing muscle to utilize carbohydrate more than fat. AB - To understand the function of the feeding-stimulatory peptide, galanin (GAL), in eating and body weight regulation, the present experiments tested the effects of both acute and chronic injections of this peptide into the paraventricular nucleus (PVN) of rats. With food absent during the test, acute injection of GAL (300 pmol/0.3 microl) significantly increased phosphofructokinase activity in muscle, suggesting enhanced capacity to metabolize carbohydrate, and reduced circulating glucose levels. It also decreased beta-hydroxyacyl-CoA dehydrogenase activity in muscle, indicating reduced fat oxidation, while increasing circulating non-esterified fatty acids (NEFA) and lipoprotein lipase activity in adipose tissue (aLPL). Chronic PVN injections of GAL (300 pmol/0.3 microl/injection) versus saline over 7-10 days significantly stimulated daily caloric intake and increased the weight of four dissected fat depots by 30-40%. These effects, accompanied by elevated levels of leptin, triglycerides, NEFA and aLPL activity, were evident only in rats on a diet with at least 35% fat. Thus, by favoring carbohydrate over fat metabolism in muscle and reversing hyperglycemia, PVN GAL may have a function in counteracting the metabolic disturbances induced by a high-fat diet. As a consequence of these actions, GAL can promote the partitioning of lipids away from oxidation in muscle towards storage in adipose tissue. PMID- 15893856 TI - Alteration of protein kinase C conformation in red blood cells: a potential marker for Alzheimer's disease but not for Parkinson's disease. AB - There is a growing evidence of early changes of blood cells in Alzheimer's disease (AD). We have developed an original novel method for quantifying the alteration of protein kinase C (PKC) by its fluorescence spectrum: by using Fim 1, a specific fluorescent probe made for protein kinase C that detects the conformational changes of this. We show that the PKC conformation is altered in red blood cells (RBC) from AD patients as compared to RBC from healthy controls. This alteration is independent of the patient's age and of the stage of the disease. It is not observed in the RBC of non-demented patients suffering from Parkinson's disease (PD). If PKC alteration is proven to be specific to AD as compared with other dementia, this method could be for a simple, low cost screening test among patients suspected of having AD and may have a strong predictive value. PMID- 15893857 TI - To screen or not to screen for adult malnutrition? AB - BACKGROUND: There is some controversy about whether all adults receiving healthcare should be routinely screened for nutritional problems. METHODS: (i) A systematic review examined the proposition that malnutrition is under-recognised and under-treated, and that nutritional interventions in malnourished patients, identified through a screening procedure produce clinical benefits (assessed using randomised controlled trials, RCTs). (ii) A systematic review of nutritional screening interventions in populations of malnourished and well nourished subjects (RCTs and non-RCTs). RESULTS: (i) The prevalence of malnutrition varies according to the criteria used, but is estimated to affect 10 60% of patients in hospital and nursing homes, 10% or more of older free-living subjects, and less than 5% of younger adults. In the absence of formal screening procedures, more than half the patients at risk of malnutrition in various settings do not appear to be recognised and/or are not referred for treatment. RCTs show that nutritional interventions in malnourished patients produce various clinical benefits. (ii) Interventions with nutritional screening in different care settings also generally suggest clinical benefits, but some are limited by small sample sizes and inadequate methodology. Factors that influence outcomes include validity, reliability and ease of using the screening procedure, the 'care gap' that exists between routine and desirable care and the need for other resources, which may increase or decrease following screening. CONCLUSIONS: The frequent failure to recognise and treat malnutrition, especially where it is common, is unacceptable. In such circumstances, the routine use of a simple screening procedure is recommended. Each health care setting should have a transparent policy about nutritional screening, which may vary according to the 'care gap', available resources, and specific populations of patients, in which the prevalence of malnutrition may vary widely. PMID- 15893858 TI - Gamma interferon levels and antibody production induced by two PvMSP-1 recombinant polypeptides are associated with protective immunity against P.vivax in Aotus monkeys. AB - Effector mechanisms responsible for providing protective immunity against Plasmodium vivax (Pv) infection were examined in Aotus monkeys vaccinated with two Pv Merozoite Surface Protein-1 (PvMSP-1) recombinant polypeptides that had previously been shown to protect vaccines against parasite challenge. Vaccine efficacy was reproducible in this trial, showing that one out of the five monkeys immunised with the recombinant protein mixture was partially protected while three others controlled parasitaemia. Antibodies reactive to the parasite's native proteins, the recombinant polypeptides and peptides spanning both recombinant fragments were detected in most vaccinees. Despite substantial Peripheral Blood Mononuclear Cell (PBMC) antigen-specific cellular proliferation not being detected, high rPvMSP-1(20) specific gamma interferon (IFN-gamma) production was found in the three animals that controlled parasitaemia. Altogether the results suggest that antibody titres and antigen-specific IFN gamma production mediate protective immunity against P. vivax. PMID- 15893859 TI - Alternative codon usage of PRRS virus ORF5 gene increases eucaryotic expression of GP(5) glycoprotein and improves immune response in challenged pigs. AB - Pigs exposed to GP(5) protein of PRRSV by means of DNA immunization develop specific neutralizing and protecting antibodies. Herein, we report on the consequences of codon bias, and on the favorable outcome of the systematic replacement of native codons of PRRSV ORF5 gene with codons chosen to reflect more closely the codon preference of highly expressed mammalian genes. Therefore, a synthetic PRRSV ORF5 gene (synORF5) was constructed in which 134 nucleotide substitutions were made in comparison to wild-type gene (wtORF5), such that 59% (119) of wild-type codons were replaced with known preferable codons in mammalian cells. In vitro expression in mammalian cells of synORF5 was considerably increased comparatively to wtORF5, following infection with tetracycline inducible replication-defective human adenoviral vectors (hAdVs). After challenge inoculation, SPF pigs vaccinated twice with recombinant hAdV/synORF5 developed earlier and higher antibody titers, including virus neutralizing antibodies to GP(5) than pigs vaccinated with hAdV/wtORF5. Data obtained from animal inoculation studies suggest direct correlation between expression levels of immunogenic structural viral proteins and immune response. PMID- 15893860 TI - Preparedness of households and catering establishments for incidents involving radioactive contamination. AB - This short paper describes a number of investigations carried out to ensure preparedness for crises involving radioactivity to catering operations and private households in Finland. The specific recommendations for catering kitchens during crises were published in 1994. A study to determine the level of adherence to these recommendations is summarised here, together with its findings and subsequent recommendations. Another study on the pre-planning of crisis menus is described. New challenges for the catering kitchens are touched upon. A crisis food preparation booklet for households is described and based on consumers' attitudes suggestions are made for how this can be improved in the future. PMID- 15893861 TI - Workshop to extend the involvement of stakeholders in decisions on restoration management. PMID- 15893862 TI - Empirical evaluation of the assumptions in identifying evidence based treatments in mental health. AB - Extensive analyses of data from the remarkably comprehensive data set established by the Treatment of Depression Collaborative Research Program (TDCRP), initiated and conducted by the National Institute of Mental Health (NIMH), enabled us to examine the contributions of three dimensions of the treatment process (type of treatment, aspects of the therapeutic relationship, and patients' pretreatment personality characteristics) to three assessments of therapeutic change (symptom reduction, reduction of vulnerability, and development of adaptive capacities) evaluated at termination and extended follow-up. The most consistent factors predicting therapeutic gain were the quality of the therapeutic relationship and patients' pretreatment personality dimensions. The implications of these findings for clinical practice, training, and research are discussed. PMID- 15893864 TI - Towards a wireless patient: chronic illness, scarce care and technological innovation in the United Kingdom. AB - 'Modernization' is a key health policy objective in the UK. It extends across a range of public service delivery and organizational contexts, and also means there are radical changes in perspective on professional behaviour and practice. New information and communications technologies have been seen as one of the key mechanisms by which these changes can be engendered. In particular, massive investment in information technologies promises the rapid distribution and deployment of patient-centred information across internal organizational boundaries. While the National Health Service (NHS) sits on the edge of a pound sterling 6 billion investment in electronic patient records, other technologies find their status as innovative vehicles for professional behaviour change and service delivery in question. In this paper, we consider the ways that telemedicine and telehealthcare systems have been constructed first as a field of technological innovation, and more recently, as management solutions to problems around the distribution of health care. We use NHS responses to chronic illness as a medium for understanding these shifts. In particular, we draw attention to the shifting definitions of 'innovation' and to the ways that these shifts define a move away from notions of technological advance towards management control. PMID- 15893865 TI - Are we employing the most effective CA 125 and CA 19-9 cut-off values to detect endometriosis? PMID- 15893866 TI - Evaluation of monocyte chemotactic protein-1 levels in peripheral blood of infertile women with endometriosis. AB - OBJECTIVE: To test whether serum monocyte chemotactic protein-1 (MCP-1) chemokine levels correlate with endometriosis in infertile women. STUDY DESIGN: A group of women with endometriosis (n = 18, infertile) was compared with patients with uterine leiomyoma (n = 16, fertile), unexplained infertility (n = 5, infertile), and healthy women (n = 16, fertile). MCP-1 expression levels were evaluated by ELISA assay. The data obtained were statistically analyzed using the Mann-Whitney test. P-Values <0.05 were considered as significant. RESULTS: MCP-1 concentrations (median; range of values) in serum were as follows: women with endometriosis (221; 101-635 pg/ml), women with unexplained infertility (167, 114 234 pg/ml), women with uterine leiomyoma (137; 88-200 pg/ml), and healthy donors (123; 98-194 pg/ml). Significant differences were observed in the women with endometriosis compared with those with uterine leiomyoma (p = 0.02) and healthy donors (p = 0.002). Among the women with endometriosis, the level of significance in MCP-1 level at rAFS stages III-IV was higher than that at rAFS stages I-II compared with healthy donors and women with leiomyoma (p = 0.002 and p = 0.02, respectively). CONCLUSIONS: These data show that an increased level of MCP-1 can characterize infertile women with endometriosis. However, further studies are needed to be able to determine whether increased MCP-1 chemokine expression can be related to infertility or is a result of endometriosis progress. PMID- 15893867 TI - Evaluation of natural coagulation inhibitor levels in various hypertensive states of pregnancy. AB - OBJECTIVE: To evaluate the role of natural coagulation inhibitors in various classifications of pregnancy associated hypertension in Turkish population living in Trakya region of Turkey. STUDY DESIGN: Serum uric acid levels, plasma protein C (PC), protein S (PS), antithrombin III (AT III) activities and activated protein C resistance (APCR) were measured in 80 pregnant women with hypertension (preeclampsia, n = 32; severe preeclampsia, n = 25; eclampsia, n = 14; chronic hypertension, n = 9) and 58 healthy pregnant women. Tukey and Tamhane multiple comparison tests, Kruskal-Wallis, chi2 and Fisher's exact tests were performed for comparison of means and/or medians. RESULTS: Serum uric acid levels were significantly elevated in women with preeclampsia and severe preeclampsia, but PS activity decreased in women with severe preeclampsia (33.2 +/- 18.9% versus 50.4 +/- 22.7%, p = 0.015) and chronic hypertension (29.5 +/- 14.5% versus 50.4+ /- 22.7%, p = 0.045) compared to healthy controls. There was no significant difference in APCR, and PC or AT III activity between the groups. Platelet counts were significantly lower in women with severe preeclampsia, compared to controls and women with chronic hypertension. CONCLUSION(S): Serum uric acid levels and plasma protein S activity may be useful as indices of severity of pathology in pregnancy associated hypertension. PMID- 15893868 TI - Effect of activin A on tumor necrosis factor-alpha/intercellular adhesion molecule-1 pathway in endometrial stromal cells. AB - OBJECTIVE[S]: Activin A and inhibin A are growth factors expressed by human endometrium involved in the control of endometrial functions. In the present study we investigated the effects of activin A and inhibin A in modulating the tumor necrosis factor (TNF)-alpha/intercellular adhesion molecule (ICAM)-1 system in cultured human endometrial stromal cells. STUDY DESIGN: Endometrial samples were obtained from 34 reproductive age women undergoing laparoscopy for benign ovarian cysts or infertility. Endometrial stromal cells were cultured and soluble ICAM-1 and TNF-alpha were measured in cell-free supernatants following treatment with or without activin A or inhibin A. Cell surface ICAM-1 was assayed by flow cytometry by staining endometrial cells with specific monoclonal antibodies. RESULTS: Activin A and inhibin A did not influence either the expression of cell surface ICAM-1 or soluble ICAM-1 shedding by cultured endometrial cells. On the other hand, TNF-alpha secretion significantly increased in presence of activin A but not of inhibin A. CONCLUSIONS: Since TNF-alpha modulates several endometrial processes such as menstruation, proliferation, apoptosis, implantation and decidualization, an effect of activin A in the physiological control of endometrium is further supported by the present data. PMID- 15893869 TI - Detection of interleukin 6 mRNA by RT-PCR in vaginal secretions: association with preterm delivery and neonatal infection in women with preterm labour and intact membranes. AB - OBJECTIVE: To find a biological marker associated with preterm delivery or neonatal infection in pregnant women with preterm labour and intact membranes. STUDY DESIGN: Cervical secretions were collected from 286 women hospitalized for preterm labour with intact membranes at 24-34 weeks' gestation. The outcomes studied were delivery before 33 and 35 weeks' gestation, chorioamnionitis, and neonatal infection, and their association with the presence of IL-6 mRNA in cervical secretions as detected by RT-PCR. The other infectious markers tested were: bacterial vaginosis and fetal fibronectin in cervical secretions; serum CRP and white blood cell count. RESULTS: The vaginal secretions of 13 of 286 women (4.7%) contained IL-6 mRNA. The only other marker tested significantly associated with IL-6 mRNA+ was the presence of streptococcus in vaginal secretions (30.8% versus 9.4% in the IL-6+ and-groups, p = 0.03). Although the difference did not reach statistical significance (p<0.06 and 0.08, respectively), in women with IL 6 mRNA in cervical secretions we observed a tendency to give birth before 33 and 35 weeks more often than the population as a whole. This group was at higher risk of neonatal infection (38.5% versus 15.1%; p = 0.04). After adjustment for infectious risk factors, IL-6 remained significantly associated with neonatal infection (OR = 4.6, 95% CI [1.1-18.9]). The sensitivity of IL-6 mRNA for neonatal infection was 11.1%. The specificity was 96.7%. CONCLUSION: The detection of IL-6 mRNA by RT-PCR in vaginal secretions allows identification of a small group of women at high risk of neonatal infection, independently of other markers of infection. PMID- 15893870 TI - Peripartum cardiomyopathy complicated by pulmonary embolism and pulmonary hypertension. PMID- 15893871 TI - Th1/Th2 cytokines balance--yin and yang of reproductive immunology. AB - For years conception of Th2 overbalance during pregnancy has been a paradigm for immunology of reproduction, while Th1 activity has been presented as unwanted component. Studies concerning Th1/Th2 balance in physiological and complicated pregnancy have been reviewed. Th1 activity during early peri-implantation period, premature and term labour not only accompanies but even predominates over Th2 activity. Th1 activity plays important role in promotion of Th2 response, regulation of placentation process, defense against infections and initiation of delivery. Together with Th2 activity it is necessary component of immunological reactions during pregnancy, both activities being inseparable like yin and yang. So paradigm of "Th1-Th2 cooperation" is much closer to reality than "Th2 phenomenon". PMID- 15893872 TI - Factors affecting the outcome of endometrial ablation using Cavaterm plus. AB - OBJECTIVE: To study treatment outcome using the modified technique: Cavaterm plus. STUDY DESIGN: Retrospective postal questionnaire, in a large teaching hospital. One hundred and twenty-eight women with menorrhagia were treated between February 2001 and April 2003. Data were collected prospectively for the duration of the procedure and alternatives offered. Follow up questionnaire was distributed during November 2003 to assess menstrual status. Multiple binary logistic regression was performed to assess factors influencing success. RESULTS: The mean follow up was 72 weeks, 103 patients (80.5%) completed the questionnaire. In 26 (25.2%) cases, there were one or more important deviations from recommended procedure. Twenty (19.4%) women had procedure-related amenorrhoea, 6 (5.8%) had spotting, 35 (34%) had light, and 26 (25.2%) had moderate bleeding. Eleven (10.7%) had a hysterectomy. The risk of failure was inversely related to age (OR 0.778, 95% CI 0.669-0.905), was higher in women who prior to surgery had longer duration of bleeding (OR 1.29, 95% CI 1.1-1.52), and when recommended selection or operative procedures were not followed (OR 5.056, 95% CI 1.097-23.3). CONCLUSION: Cavaterm plus is associated with high patient satisfaction. The technique remains a good choice for women wishing to avoid hysterectomy, but there is a need to observe determinants of poor outcome. PMID- 15893873 TI - Interactions of cytochrome c with DNA at glassy carbon surface. AB - Cyclic voltammetry (CV) was used to investigate the interactions of Cytochrome c (Cyt c) with deoxyribonucleic acid (DNA) at glassy carbon (GC) electrodes. The results indicate that there are strong interactions between Cyt c and DNA. The binding constant (k(A)) and binding free energy (Delta(r)G) of Cyt c with dsDNA are (1.69+/-0.38) x 10(5) L.mol(-1) and -(29.76+/-0.56) kJ.mol(-1), respectively; and those of Cyt c with ssDNA are (3.35+/-0.50) x 10(5) L.mol(-1) and -(31.49+/ 0.37) kJ.mol(-1), respectively. The binding sites are achieved to be 3.3 bp per Cyt c molecule with dsDNA and 4.0 nucleotides (ssDNA) binding one Cyt c molecule. This experiment affords a valid method for investigating the interactions between DNA and proteins by electrochemical techniques. PMID- 15893874 TI - Intracranial lipomas--a clinical study. AB - OBJECTIVE: This study aimed to investigate the localization of the lipoma, as well as associated intracranial and extracranial lesions in 14 patients immediately following hospital admission. The pathological findings from the neurological examinations of these patients are also investigated. METHODS: Fourteen patients who were admitted to our clinic with a variety of symptoms and diagnosed with intracranial lipoma were included in the study. Problems presented upon admission, neurological findings, and other existing system abnormalities were evaluated. Localization of the lipomas and accompanying pathologies were determined by using computerised tomography and magnetic resonance imaging. RESULTS: The most frequent reasons for admission of patients with intracranial lipomas were: headache 7 (50%), trauma 3 (21.5%), epilepsy 3 (21.5%) and one with symptoms due to the local mass effect of tumor (7%). Although the pericallosal region is accepted as the region where lipomas commonly occur, this study found the most frequent occurrence in the quadrigeminal cistern. Intracranial lipoma calcification was only evident in 1 of the 14 patients. In addition, contrary to the expectations, intracranial and extracranial lesions accompanying lipomas were rare. All patients received systematic treatment. CONCLUSION: This study showed that intracranial lipomas are more frequent in the quadrigeminal region of the brain; most are asymptomatic, generally caught incidentally; and accompanying intracranial and extracranial pathologies are less common than expected. PMID- 15893875 TI - The role of Rac1 in maintaining malignant phenotype of mouse skin tumor cells. AB - We have previously developed an in vitro tumor progression model with mouse skin keratinocytes to study the molecular targets that mediate the tumor cell's progression from a benign to a malignant phenotype. The malignantly transformed cells were found to have elevated MAP kinase signaling and increases in AP-1, NFkappaB and cAMP response element (CRE) transcription factors activities compared to their benign counter-part. In this study, we showed that Rac1, a member of the Rho superfamily of small GTPases, functions as a key signaling molecule that mediates these malignant phenotypes. We used a doxycycline inducible system to express dominant negative Rac1 (N17 Rac1) in the squamous cell carcinomas producing 6M90 cell line. Conditional expression of the N17 Rac1 was able to decrease multiple markers of malignancy including: growth rate, colony formation, migration, invasion and most importantly, in vivo tumor growth. In addition, these phenotypic changes were accompanied by decreases in mitogenic signals, which include ERK1/2, JNK, and PI-3 kinase/Akt activation. Transactivation mediated by AP-1, NFkappaB, and CRE were also attenuated by expression of dominant negative Rac1. These observations led us to conclude that Rac1 signaling is required for the malignant phenotypes of the squamous cell carcinoma cells. PMID- 15893876 TI - Cerebral interstitial levels of glutamate and glutamine after intravenous administration of nutritional amino acids in neurointensive care patients. AB - After severe trauma or disease glutamine (GLN) is mobilised from all muscles, including the heart and smooth muscles. The result is weakness and fatigue which affects recovery. The breakdown of muscle tissue can be counteracted by external GLN supply. There are concerns, however, that increasing the blood glutamine (Blood-GLN) concentration in patients with acute brain diseases is harmful by elevating the CNS interstitial (IS) concentration of glutamate (CNS-GLT), and that this may result in a secondary excitotoxic injury. We therefore studied the IS CNS-GLN and CNS-GLT when a commercially available nutritional amino acid solution was given intravenously. Ten NICU patients were included. The IS concentrations of amino acids in the brain were measured using intracerebral microdialysis. Blood concentrations of amino acids were measured before and after the amino acid infusion. The change in Blood-GLN was 2.14 (median; range 1.34 3.22) times the basal levels and Blood-GLT increased 1.37 (median; range 0.93 3.45) times basal levels. Both changes were statistically significant. The changes in CNS-GLN was 1.21 (median; range 0.72-1.92) and for CNS-GLT 0.96 (median; range 0.45-1.53) times the basal levels. This was statistically significant for CNS-GLN but not for CNS-GLT. A high initial CNS-GLT (55.3 micromol/l) in one patient increased even further to 84.4 micromol/l after infusion of amino acid solution. We submit that nutritional amino acid solutions can be administrated to some patients with acute brain disease without increasing the CNS-GLT values. However, since BBB function was not quantified in our study, further evaluation of this issue is warranted. PMID- 15893877 TI - When word category information encounters morphosyntax: an ERP study. AB - The present study investigated the relationship between two different syntactic information types, namely word category and morphosyntax. The event-related brain potential (ERP) pattern of acoustically presented sentences containing two syntactic anomalies (word category and subject-verb agreement) was compared to the ERP response to sentences containing a single violation. The ERPs for the agreement violation revealed a left anterior negativity (LAN) indicating the detection of the morphosyntactic error, followed by a P600 reflecting processes of reanalysis. The ERPs for both the category and the combined violation showed an early negativity reflecting processes of phrase structure building, followed by a P600 indicating syntactic reanalysis. Additionally, a broadly distributed negativity following the early negativity and preceding the P600 was observed. This ERP component is suggested to reflect reference specification processes arising from the specific sentence structure used in the present study. The ERP pattern for the combined violation suggests no additivity or interaction between the two syntactic anomalies in the early time windows (early negativity, reference-related negativity, and LAN), whereas interactive effects are observed in a late time range (P600). PMID- 15893878 TI - Glucose-derived Amadori compounds of glutathione. AB - Under the chromatographic conditions used in these studies we observed time- and concentration-dependent formation of N-1-Deoxy-fructos-1-yl glutathione as the major glycation product formed in the mixtures of GSH with glucose. N-1-Deoxy fructos-1-yl glutathione had a characteristic positively charged ion with m/z=470 Th in its LC-MS spectra. Mixtures of glutathione disulfide and glucose generated two compounds: N-1-Deoxy-fructos-1-yl GSSG (m/z=775 Th) as major adduct and bis di-N, N'-1-Deoxy-fructos-1-yl GSSG (m/z=937 Th) as the minor one. All three compounds showed a resonance signal at 55.2 ppm in the 13C-NMR spectra as C1 methylene group of deoxyfructosyl, which represents direct evidence that they are Amadori compounds. All three compounds purified from GSSG/Glc or GSH/Glc mixtures also showed LC-MS/MS fragmentation patterns identical to those of the synthetically synthesized N-1-Deoxy-fructos-1-yl glutathione, N-1-Deoxy-fructos-1 yl GSSG and bis di-N, N'-1-Deoxy-fructos-1-yl GSSG. N-1-Deoxy-fructos-1-yl glutathione was shown to be a poor substrate for glutathione peroxidase (6.7% of the enzyme's original specific activity) and glutathione-S-transferase (25.7% of the original enzyme's specific activity). Glutathione reductase failed to recycle the disulfide bond within the structure of di-substituted bis di-N, N'-1-Deoxy fructos-1-yl GSSG. It showed only 1% of the original enzyme's specific activity, but retained its ability to reduce the disulfide bond within the structure of N-1 Deoxy-fructos-1-yl GSSG by 57% of its original specific activity. Since the GSH concentration in diabetic lens is significantly decreased and the glucose concentration can increase 10-fold and higher, the formation of Amadori products of the different forms of glutathione with this monosaccharide may be favored under these conditions and could contribute to a lowering of glutathione levels and an increase of oxidative stress observed in diabetic lens. PMID- 15893879 TI - Regulation of SNARK activity in response to cellular stresses. AB - SNARK is a member of the AMPK subfamily of serine/threonine protein kinases. In this study, we examined the regulation of SNARK activity in kidney (BHK, HEK293), pancreatic beta-cell insulinoma (INS-1), hepatocarcinoma (H4IIE) and keratinocyte (NRKC)-derived cell lines in response to diverse cellular stresses. We show that SNARK activity is regulated by glucose- or glutamine-deprivation, induction of endoplasmic reticulum stress by homocysteine or DTT, elevation of cellular AMP and/or depletion of ATP, hyperosmotic stress, salt stress, ultraviolet B radiation and oxidative stress caused by hydrogen peroxide. Moreover, the regulation of SNARK activity in response to cellular stresses depends greatly upon cell type. Furthermore, SNARK activity is downregulated by metformin in a dose- and time-dependent manner in H4IIE cells. These observations support a role for SNARK as a molecular component of the cellular stress response. PMID- 15893880 TI - Effects of light and chloramphenicol stress on incorporation of nitrogen into cyanophycin in Synechocystis sp. strain PCC 6308. AB - (1)H NMR spectroscopy was used to compare the uptake of nitrogen into cyanobacterial cyanophycin from two sources: from the breakdown of intracellular proteins and amino acids, and directly from the external growth medium. Cells grown initially in medium containing (14)N-nitrate were transferred to (15)N nitrate medium in the presence of chloramphenicol in both low (4 microE m(-2) s( 1)) and normal (100 microE m(-2) s(-1)) light, and in low light alone. Cyanophycin was separated from cells and analyzed by (1)H NMR spectroscopy. Cyanophycin is synthesized both from (14)N (degradation of cellular proteins) and from (15)N in the medium, the latter at a faster rate and to a greater extent under all conditions. SDS-PAGE showed that cyanophycin synthesis takes place by addition of monomers to already synthesized polymer. PMID- 15893881 TI - Up-regulated phosphorylation of signal transducer and activator of transcription 3 and cyclic AMP-responsive element binding protein by peripheral inflammation in primary afferent neurons possibly through oncostatin M receptor. AB - Oncostatin M (OSM), a member of interleukin-6 family cytokines, contributes to the development of nociceptive sensory neurons. However, little is known about the role of OSM in dorsal root ganglia (DRGs) of adult mice after peripheral inflammation. In the present study, we showed that OSM mRNA was highly expressed in the inflamed skin during acute inflammation induced by complete Freund's adjuvant (CFA), while the expression of oncostatin M receptor (OSMR) did not change in the ipsilateral DRG. Although peripheral inflammation induced significant increases in the number of neurons with phosphorylated extracellular signal-regulated kinase (p-ERK) and phosphorylated p38 mitogen-activated protein kinase (p-p38) in ipsilateral DRGs, OSMR-positive neurons exhibited neither p-ERK nor p-p38. In addition, we found significant increases in the number of neurons with phosphorylated signal transducer and activator of transcription 3 (p-STAT3) and phosphorylated cAMP-responsive element binding protein (p-CREB) in the ipsilateral DRGs. Interestingly, OSMR-positive neurons with p-STAT3 and p-CREB were significantly increased after peripheral inflammation. Thus, our results suggest that acute inflammation induce the phosphorylations of several signal molecules, including ERK, p38, cAMP-responsive element binding protein, and STAT3. Among them, the up-regulation of p-STAT3 and p-CREB may be induced possibly through OSMR. PMID- 15893882 TI - Pigment epithelium-derived factor induces pro-survival genes through cyclic AMP responsive element binding protein and nuclear factor kappa B activation in rat cultured cerebellar granule cells: Implication for its neuroprotective effect. AB - Pigment epithelium-derived factor (PEDF) protects immature cerebellar granule cell neurons (CGCs) against apoptosis induced by K+ and serum deprivation. However, the precise mechanism of this protection remains unknown. We recently reported that the transcription factor nuclear factor kappa B (NF-kappaB) is activated in PEDF-treated CGCs. Although it is well known that NF-kappaB blocks apoptotic cell death through the induction of pro-survival factors, the effects of PEDF on the expression of these factors are not fully understood. In this study, we employed the use of reverse transcriptase-polymerase chain reaction to analyze the gene expression of certain pro-survival genes and found that genes such as c-IAP1, c-IAP2, FLIPs, A1/Bfl-1 and Mn-SOD were induced in PEDF-treated neurons. On the other hand, no induction was observed of the pro-apoptotic Bcl-2 family members Bax and Bid at any time from 3 to 24 h following PEDF addition. Furthermore, phosphorylation of cyclic AMP-responsive element binding protein (CREB) and increment of nuclear cyclic AMP-response element (CRE)-like DNA binding were observed in PEDF-treated CGCs. The anti-apoptotic effect of PEDF was blocked by overexpression of dominant negative CREB or a mutated form of IkappaBalpha. These results suggested that induction of both CRE- and NF-kappaB dependent genes is required for the observed neuroprotective effects of PEDF on CGCs. PMID- 15893883 TI - Dexamethasone modulates the development of morphine tolerance and expression of glutamate transporters in rats. AB - We recently demonstrated an increase in spinal cerebrospinal fluid (CSF) excitatory amino acids (EAAs) in morphine-tolerant rats after morphine challenge. The present study examined whether co-infusion of the glucocorticoid dexamethasone (DEX) co-infusion inhibited morphine tolerance and the morphine challenge-induced EAAs increase after long-term morphine infusion. Intrathecal (i.t.) catheters and one microdialysis probe were implanted to male Wistar rats. Rats were divided into four groups: i.t. morphine (15 microg/h), saline (1 microl/h), DEX (2 microg/h), or DEX (2 microg/h) plus morphine (15 microg/h) infusion for 5 days. Tail-flick responses were examined before drug infusion and daily after the start of infusion for 5 days. Moreover, on day 5 after morphine challenge (50 microg, i.t.), CSF EAAs was also measured. Rat spinal cords were removed on day 5, and prepared for Western blot analysis of different glutamate transporters (GTs). The AD50 (analgesic dose) on day 5 was 1.33 microg in saline infused rats, 83.84 microg in morphine-tolerant rats, and 10.15 microg in DEX plus morphine co-infused rats. Single DEX (2 microg, i.t.) injection did not enhance morphine's antinociceptive effect in either naive or morphine-tolerant rats. No difference in CSF EAA level was observed in all groups between baseline (before drug infusion) and on day 5 after tolerance developed. Surprisingly, on day 5, after morphine challenge, an increase in glutamate and aspartate (284+/ 47% and 201+/-18% of basal) concentration was observed, and morphine lost its antinociceptive effect (maximum percent effect, MPE = 41+/-12%), whereas DEX/morphine co-infusion inhibited morphine-evoked EAA increase with a MPE = 97+/ 2%. DEX co-infusion prevented the downregulation of glial glutamate transporters (GLAST (Glu-Asp transporter) and GLT-1 (Glu transporter-1)), but not the neuronal GT EAAC1 (excitatory amino acid carrier). Upregulation of GLT-1 was also observed (204+/-20% of basal). DEX co-infusion inhibits the morphine-challenge induced EAA increase and prevents the loss of morphine's antinociceptive effect after long term morphine infusion. PMID- 15893884 TI - Cyclic AMP response element-binding protein is required for normal maternal nurturing behavior. AB - Analysis of mice with targeted disruptions of fosB or the gene encoding dopamine beta-hydroxylase suggests that FosB and adrenergic signaling play critical roles in maternal nurturing behavior. The majority of neonates born to null females from either mutation fail to thrive, and virgin mutant females of both lines exhibit impaired pup retrieval. Considering whether FosB and adrenergic signaling might share a signaling pathway important for maternal behavior, we examined the role of a potential intermediary, cyclic AMP response element-binding protein (CREB). Here we report that approximately 40% of neonates (all heterozygous) born to mice lacking the major isoforms of CREB (Creb-alphaDelta-/-) died within several days of birth. In contrast, heterozygotes born to Creb-alphaDelta+/- females thrived. Cross-fostering demonstrated that neonates born to Creb alphaDelta(-/dagger/-) females thrived when reared by wild-type females, and that Creb-alphaDelta-/- females were capable of rearing neonates whose maternal care was initiated by wild-type females. Further, virgin Creb-alphaDelta-/- females were deficient in pup retrieval despite exhibiting normal investigation of pups and of novel objects. No maternal behavior phenotype was present in mice with a null mutation of the cyclic AMP response element modulator (Crem) gene. Interestingly, the number of cells immunostaining for phospho-CREB (on Ser(133)) in the medial preoptic area of the hypothalamus, a key region for the expression of maternal behavior, increased nearly three-fold in wild-type mice following exposure to pups but not to novel objects. On the other hand, basal expression and induction of FosB in response to pup exposure appeared to be independent of CREB because levels were equivalent between wild-type and Creb-alphaDelta-/- females. These results implicate CREB in maternal nurturing behavior and suggest that CREB is not critical for expression or induction of FosB in adult virgin female mice. PMID- 15893885 TI - Components and antibacterial activity of the roots of Salvia jaminiana. AB - The acetone extract of the roots of Salvia jaminiana, containing the sterols campestanol, stigmasterol and sitosterol, and five known diterpenoids, ferruginol, cryptanol, 6,7-dehydroroyleanon, 6-hydroxysalvinolone and microstegiol, remarkably inhibited the growth of Bacillus subtilis, Staphylococcus aureus ATCC 25923 and Streptococcus alpha-hemolitic. PMID- 15893886 TI - A new dihydroxysterol from the marine phytoplankton Diacronema sp. AB - Diacronema sp. was cultured and its sterols were separated by column chromatography on silica gel. The new sterol 24-ethyl-4alpha-methyl-cholestane 3,20-diol (1) was characterised by NMR and MS spectrometry, as well as (22E)-24 ethyl-4alpha-methyl-5alpha-cholest-22-en-3beta-ol (2) and beta-sitosterol, the major components of the sterol fractions. Neither the biosynthetic origin of the new dihydroxysterol nor its role in the biochemistry of Diacronema is known. PMID- 15893887 TI - Antibacterial effects of commercial essential oils over locally prevalent pathogenic strains in Mexico. AB - Locally prevalent pathogenic bacteria 189 Gram (-) and 135 Gram (+) strains, all isolated from pediatric patients severely infected, were tested in vitro against 11 essential oils from commercial origin. All the strains showed resistance to selected antibiotics. Cinnamomum verum, Origanum vulgare and Thymus vulgaris exhibited the highest and broadest antibacterial activity. Emphasis is made in the potential implications of these resources, uncommon at the clinical setting of the study, employed against non-commercial, locally pathogenic strains, being a step to submit in the ensuing period essential oils from plants used in Mexican traditional medicine. PMID- 15893888 TI - Antimicrobial activity of Viola tricolor herb. AB - The antimicrobial activity of infusion, decoction, ethanol extract and fractions obtained by successive extraction of Viola tricolor herb with dichloromethane, ethyl acetate and methanol was evaluated. The infusion, decoction and ethanol extract were found to be most effective against the tested microorganisms. PMID- 15893889 TI - Ethanol-induced FOS immunoreactivity in the brain of mu-opioid receptor knockout mice. AB - Using mu-opioid receptor knockout (MKO) mice, we examined ethanol-induced FOS immunoreactivity (FOSir) in the brain as an indicator of neuronal activation to assess the role of the mu-opioid receptor in modulating ethanol's actions in the central nervous system (CNS). Saline stimulated FOSir in the paraventricular thalamic nucleus (PVA) and the dorsal hypothalamic area (DA) in MKO mice, but not in wild-type (WT), suggesting that MKO homozygotes may differ responsively from WT. Treatment with ethanol (4 g/kg, i.p.) induced FOSir in the PVA, DA, supraoptic (SO), paraventricular hypothalamic (PVN), lateral parabrachial (LPB), locus coeruleus (LC) and Edinger-Westphal (EW) nuclei in both MKO and WT mice. However, ethanol stimulated modest FOSir in the lateral septal division (LSV), suprachiasmatic nucleus (SCh) and the dorsal and ventral lateral geniculate nuclei (DLG and VLG) in WT mice, but not in MKO mice. In contrast, higher levels of ethanol-induced FOSir were observed in the ventral pallidum (VP) and globus pallidus (GP) of MKO mice as compared to WT. These data suggest that ethanol continues to activate several brain regions, even without the mu-opioid receptor pathway. However, the mu-opioid receptor may be significant in mediating ethanol's effects in some restricted areas of the brain. PMID- 15893890 TI - Interaction between yeast mitochondrial and nuclear genomes: null alleles of RTG genes affect resistance to the alkaloid lycorine in rho0 petites of Saccharomyces cerevisiae. AB - Some nuclear genes in Saccharomyces cerevisiae (S. cerevisiae) respond to signals from the mitochondria in a process called by Butow (Cell Death Differ. 9 (2002) 1043-1045) retrograde regulation. Expression of these genes is activated in cells lacking mitochondrial function by involvement of RTG1, RTG2 and RTG3 genes whose protein products bind to "R-boxes" in the promoter region; RTG2p is a cytoplasmic protein. Since S. cerevisiae rho0 strains, lacking the entire mitochondrial genome, are resistant to lycorine, an alkaloid extracted from Amaryllis plants, it could be hypothesized that in rho0 cells the dysfunctional mitochondrial status stimulates overexpression of nuclear genes very likely involved in both nuclear and mitochondrial DNA replication. In this report we show that the resistance of rho0 cells to lycorine is affected by the deletion of RTG genes. PMID- 15893891 TI - Topical exposure to carbon disulfide induces epidermal permeability alterations in physiological and pathological changes. AB - Carbon disulfide (CS2) has been suggested its possible skin toxicity. Neither a dose-response relationship nor any mechanism of CS2-exposure regarding epidermal permeability alterations has been postulated. The objectives of this study were to evaluate the dose-dependent association and the pathological changes with CS2 topically applied to mouse epidermis. Four concentrations of CS2 (0% (controls), 10%, 15%, and 20% in ethanol) were topically applied to a 1.8 cm2 area of the lateral abdomen of female nude mice for 10 min. Time-series transepidermal water loss (TEWL) profile, morphological examinations by both light microscopy (hematoxylin/eosin stain and Nile Red stain) and electronic microscopy, and lipid analysis by high performance thin-layer chromatography (HPTLC) were used to evaluate the epidermal impairment. We found no recovery occurred within 72 h exposure to 20% CS2 in contrast to substantial recovery found in 10% and 15% CS2 exposure. Clear dose-dependent fashions were shown in TEWL elevations, recovery retardation, and lipid extraction across the ethanol (control), 10%, 15%, and 20% CS2 exposures. Two mechanistic pathways were raised to account for CS2-induced epidermal alterations: intercellular lipid depletion and keratinocyte damage. A study with different test animal species is warranted owing to the discrepancies in epidermis between nude mice and other species. PMID- 15893892 TI - Serological status for N. caninum, bovine viral diarrhea virus, and infectious bovine rhinotracheitis virus at pregnancy testing and reproductive performance in beef herds. AB - Neospora caninum, infectious bovine rhinotracheitis (IBR), and bovine viral diarrhea virus (BVDV) are important differentials for the diagnosis of infectious reproductive loss in beef herds. The objective of this study was to describe the serological status of both pregnant and non-pregnant beef cows from herds with varying levels of reproductive success. The study provided an opportunity to examine whether there were any associations between serological status for BVDV, IBR, and N. caninum and pregnancy status, as well as the subsequent risk of abortion, or stillbirth. Samples were collected from 2516 cows and heifers from 66 herds; 31 herds where the proportion pregnant was <90% and 35 randomly selected herds where the proportion pregnant was > or =90%. Of these samples 5.9% were positive for antibodies to N. caninum, 20.4% had titres >1:80 to IBR, 91.8% had titres > or =1:256 to BVDV type 1, and 23.9% had titres > or =1:256 to BVDV type 2. N. caninum antibody concentration was associated with an increased individual animal risk of non-pregnancy (OR(logS/P), 1.9; 95% CI, 1.2-2.9) and abortion (OR(pos/neg), 2.8; 95% CI, 1.1-7.5). The proportion of animals at pregnancy testing with antibodies to BVDV type 2 above 1:3000 (OR(10%changeinprevalence), 2.3; 95% CI, 1.5-3.5) was also associated with an increased risk of abortion. No other measures of antibody status were associated with reduced reproductive performance in this group of herds. Antibodies to Mycobacterium avium spp.paratuberculosis were also measured; 0.7% of samples were positive (sample to positive (S/P) >0.25) and 3.6% were suspicious (S/P, 0.10 0.25). PMID- 15893893 TI - Topical absorption of piroxicam from organogels--in vitro and in vivo correlations. AB - In view of their good skin tolerability, glyceryl fatty acid esters were used as organogelators, and their effects in the topical penetration of piroxicam (Px) were investigated. The in vivo skin penetration was evaluated by measuring the anti-inflammatory effect in rats, where we found that Px incorporated into glyceryl fatty acid ester organogels exhibited a significantly greater inhibition of oedema than that of the placebo control either when applied locally (p < 0.001), or via transdermal absorption (p < 0.01 and < 0.05, respectively). As the Px concentration was increased, the extent of oedema inhibition rose in accordance with a power law. Comparisons with traditional galenic organogels and a marketed product revealed that the relative biological availability of Px was better from glyceryl fatty acid ester organogels, except when calculated for D1 versus T2 and T3. In order to predict the extent of in vivo skin absorption, we measured the penetration coefficient and the in vitro penetration. In accordance with theory, the extent of in vivo oedema inhibition increased as P(oct/w) increased, and maximum inhibition was observed at logP = 2.0211. However, the in vitro penetration through a synthetic membrane did not correlate with the in vivo results, the reason for which might be the different natures of the model barriers. PMID- 15893894 TI - Impurity formation studies with peptide-loaded polymeric microspheres Part I. In vivo evaluation. AB - The purpose of the present investigation was to assess the peptide related substances or impurities formed during incubation of drug loaded poly-(D,L lactide-co-glycolide) (PLGA) and poly-(D,L-lactide) (PLA) microspheres under in vivo conditions. Sprague-Dawley rats were injected with separate batches of octreotide microspheres prepared by either an oil/water or oil/oil dispersion technique. At specified time points (days 14, 22, 30, and 41), animals were sacrificed and microsphere particles were recovered from the subcutaneous injection sites. The recovered particles were further extracted with 1:1 mixture of dimethylsulfoxide:dichloromethane for subsequent impurity analysis by HPLC and mass spectrometry. During incubation, the percentage purity of parent compound depended on the PLGA co-monomer ratio (e.g. 50:50, 85:15, and 100:0 glycolide:lactide ratios). After 41 days of incubation, for instance, octreotide area percentage by HPLC was determined to be approximately 47% for PLGA 50:50 microspheres, approximately 75% for PLGA 85:15 microspheres, and approximately 87% for PLA microspheres. Spectral analysis of particle extracts revealed the presence of peptide related substances with 58 m/z and 72 m/z units higher than the parent peptide m/z value. This indicated the presence of glycoyl and lactoyl covalent substitutions on the drug compound, resulting from chemical interaction between peptide amine groups and PLGA or PLA ester groups. PMID- 15893895 TI - Regulatory effect of atopic allergic reaction by Carpopeltis affinis. AB - Carpopeltis affinis Okamura (CA, Halymeniaceae) has long been used as therapeutics for various allergic diseases in Korea. The precise effects of CA in experimental models, however, have remained unknown. We studied the effects of a methanol extract of CA on atopic allergic reaction. Histamine content was measured by the o-phthalaldehyde spectrofluorometric procedure. Cytokines were measured by a modified enzyme-linked immunosorbent assay. Cytotoxicity was determined by the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. CA significantly inhibited the histamine release and beta hexosaminidase release from rat peritoneal mast cells. CA also inhibited interleukin-8 and tumor necrosis factor-alpha secretion from the phorbol 12 myristate 13-acetate and A23187-induced HMC-1 cells (human mast cell line). 48 h exposure to CA (1.0, 10, and 100 microg/ml) had little effect on HMC-1 cell viability. Our results suggest that CA has an inhibitory effect on mast cell dependent allergic reaction and thus may be useful in the treatment of atopic dermatitis. PMID- 15893896 TI - Anti-inflammatory activities of flavonoids isolated from Caesalpinia pulcherrima. AB - The anti-inflammatory activities of five flavonoids, namely 5,7 dimethoxyflavanone (1), 5,7-dimethoxy-3',4'-methylenedioxyflavanone (2), isobonducellin (3), 2'-hydroxy-2,3,4',6'-tetramethoxychalcone (4) and bonducellin (5), all of them isolated from Caesalpinia pulcherrima L. was studied in lipopolysaccharide (LPS) and interferon (IFN)-gamma activated murine peritoneal macrophages. These five compounds significantly and dose-dependently inhibited the inflammatory mediators; nitric oxide (NO), and cytokines [tumor necrosis factor (TNF)-alpha and interleukin (IL)-12]. According to their inhibitory results, the order of anti-inflammatory potency was compounds 3>5>4>2>1. Furthermore, peritoneal macrophages were pre-activated with LPS/IFN-gamma for 24h, and determined the inhibitory effects of the above-mentioned isolates on the production of NO after a further 24h. The present study supports the use of Caesalpinia pulcherrima for the treatment of inflammatory diseases in traditional medicine. This is the first study on compounds 1-5 about their anti-inflammatory activities. PMID- 15893897 TI - Identification of N,N-dimethylamphetamine formed by methylation of methamphetamine in formalin-fixed liver tissue by multistage mass spectrometry. AB - Methamphetamine is methylated in the presence of unbuffered formalin solutions within hours at room temperature. The product, N,N-dimethylamphetamine, is also found in human liver exposed to methamphetamine followed by incubation with formalin. In the present study, a direct mass spectrometric method was developed to identify N,N-dimethylamphetamine in human liver before and after treatment with formalin. Human liver samples were obtained from four deaths that were investigated by the West Virginia Office of Chief Medical Examiner. Full toxicological analysis was conducted on samples from the decedents and methamphetamine was among the positive findings in each case. The method used to expose liver tissue to formaldehyde involved treating a small piece of liver from each case with formalin solution (20% v/v) for 24 h at room temperature. The formalin treated tissues were homogenized and the resulting suspension was sonicated for 5 min, and then centrifuged. Supernatant aliquots were directly analyzed by electrospray ionization (ESI) mass spectrometry without chromatographic isolation. Positive ion multistage mass spectra recorded in MS, MS/MS and MS/MS/MS (MS3) modes were used to confirm the presence of N,N dimethylamphetamine and methamphetamine in the mixture. Liver tissue not treated with formalin did not contain a detectable level of N,N-dimethylamphetamine. Decreases in methamphetamine concentrations in liver tissue resulting from treatment with formalin were measured using deuterium-labeled methamphetamine as internal standard. The method can be completed in less than 2 h on thawed tissue. The results suggest that the process of fixing tissues with formalin may lead to false negative findings for methamphetamine. PMID- 15893898 TI - The value of contrast and subtraction arthrography in the assessment of aseptic loosening of total hip prostheses: a meta-analysis. AB - OBJECTIVE: To summarize and compare the diagnostic accuracy of contrast and subtraction arthrography in the assessment of aseptic loosening of total hip arthroplasties. DESIGN: This meta-analysis was performed using methods described by the Cochrane Methods Group on Systematic Reviews of Screening and Diagnostic Tests. We included original, English-language papers published between January 1975 to October 2004 that examined contrast-enhanced arthrography with or without subtraction for diagnosis of loosening of total hip prostheses. A qualitative and quantitative analysis was performed by two investigators. RESULTS: With regard to the acetabular component, pooled sensitivity and specificity for contrast arthrography was 70% (95% confidence interval, 52-84) and 74% (95% CI, 53-87), respectively. Subtraction arthrography had a significantly higher sensitivity of 89% (95% CI, 84-93) (p=0.01), with a similar specificity of 76% (95% CI, 68-82). For the femoral component, pooled sensitivity and specificity for contrast arthrography were 63% (95% CI, 53-72) and 78% (95% CI, 68-86). Pooled estimates for subtraction arthrography revealed a significantly higher sensitivity of 86% (95% CI, 74-93) (p=0.003). Specificity was 85% (95% CI, 77-91) and was similar to the data of contrast arthrography (p=0.23). CONCLUSION: Using the present data we found that the subtraction arthrography is a sensitive technique for detection of loosening of total hip prostheses, offering added value over contrast arthrography, especially for evaluation of the femoral component. PMID- 15893899 TI - Quantitative NMR spectroscopy--applications in drug analysis. AB - NMR spectroscopy being a primary ratio method of measurement is highly suitable to evaluate the quality of drugs. NMR spectroscopy can be used for the identification of a drug substance, the identification and quantification of impurities arising from the synthesis pathway and degradation, or residual solvents as well as the determination of the content in the assay. This review gives an overview of the application of quantitative NMR spectroscopy in International Pharmacopoeias and for licensing purposes. PMID- 15893900 TI - Trends in nalidixic acid resistance in nontyphoidal Salmonella isolated from 1999 to 2002: decreased susceptibility to 6 fluoroquinolones. AB - We determined the resistance to nalidixic acid and 6 fluoroquinolones of 771 nontyphoidal Salmonella strains isolated from humans between 1999 and 2002. A total of 22 different serotypes were identified among the Salmonella isolates, the most common being Salmonella Enteritidis (79%) and Salmonella Derby (8%). Resistance to nalidixic acid increased from 38% in 1999 to 43% in 2002. This resistance was not homogeneous among the different serotypes, with the highest percentage of resistant isolates belonging to Salmonella Hadar (79%) followed by Salmonella Enteritidis (46%). Reduced ciprofloxacin susceptibility (minimal inhibitory concentrations [MICs] from 0.12 to 0.5 microg/mL) was observed in 300 (39%) Salmonella. In our study, the nalidixic acid-resistant strains had an MIC90 at least 4-fold higher than the susceptible ones for all the fluoroquinolones tested, thus showing that resistance to nalidixic acid is an indicator of low level resistance to all fluoroquinolones. PMID- 15893901 TI - Effects of voriconazole, granulocyte-macrophage colony-stimulating factor, and interferon gamma on intracellular fluconazole-resistant Candida glabrata and Candida krusei in human monocyte-derived macrophages. AB - Infections caused by fluconazole-resistant Candida glabrata and Candida krusei are increasingly common causes of morbidity and mortality. We investigated the intracellular killing of fluconazole-resistant C. glabrata and C. krusei by cytokine-activated human monocyte-derived macrophages (MDM) in the presence and absence of voriconazole. For C. glabrata, MDM were activated with either granulocyte-macrophage colony-stimulating factor (GM-CSF) or interferon gamma (IFN-gamma) before infection, after infection, or both before and after infection, whereas for C. krusei MDM were activated with cytokines both before and after infection. Activated MDM were infected, treated with voriconazole, and then lysed, and viable yeast in the lysates enumerated at 0, 24, or 48 h after infection. In the presence of voriconazole (2.5 x MIC), the best activity against C. glabrata occurred when MDM were activated with GM-CSF for 24 h before infection as well as after infection or when they were activated for 24 h before infection alone. A lesser effect was observed when MDM were activated for at least 1 h before infection or when they were treated with cytokines only after infection. IFN-gamma activation had a significant but lesser effect than GM-CSF. Activity against C. krusei in the presence of voriconazole was greatest when MDM were activated with IFN-gamma rather than GM-CSF. Our results suggest that cytokines increase the intracellular anticandidal effect of voriconazole and may be useful as therapeutic adjuvants to voriconazole for treatment of infections caused by fluconazole-resistant C. glabrata and C. krusei. PMID- 15893902 TI - Sterol uptake in Candida glabrata: rescue of sterol auxotrophic strains. AB - Candida glabrata is emerging as a more common and important human pathogen. It is less susceptible to azole antifungals than Candida albicans, thus, posing some unique treatment challenges. Previously undetected C. glabrata isolates were identified from clinical specimens by adding bile to the growth medium. Cholesterol was found to be the responsible ingredient in bile. Six bile dependent isolates were characterized and were found to exhibit wild-type equivalent growth when provided human or bovine serum or free cholesterol. Sterol profiles of the 6 isolates and a C. glabrata matching wild-type strain not requiring cholesterol indicated that 2 were defective in squalene epoxidase (encoded by the ERG1 gene) activity, 3 were defective in lanosterol synthase (encoded by the ERG7 gene) activity, and the sixth was defective in heme biosynthesis. All 7 isolates produced profiles that contained cholesterol transported from the media. Because Saccharomyces cerevisiae mutants unable to synthesize heme will take up exogenous sterol under aerobic conditions, hem1 nulls of C. glabrata and C. albicans were generated and tested for growth on ergosterol media. Only the C. glabrata hem1 was able to grow indicating significant differences in exogenous sterol uptake between the 2 organisms. The ability of C. glabrata to replace ergosterol with host sterol may be responsible for its elevated azole resistance. PMID- 15893903 TI - Reduction in turnaround time for laboratory diagnosis of pulmonary tuberculosis by routine use of a nucleic acid amplification test. AB - Every first diagnostic specimen from suspected patients with pulmonary TB was tested by a nucleic acid amplification test (NAAT) to determine the reduction in turnaround time (TAT) for detecting Mycobacterium tuberculosis (MTB) that was possible under normal laboratory operating conditions. NAAT (Gen-Probe Mycobacterium tuberculosis Direct Testtrade mark) was performed on the first specimen and liquid culture (BACTEC 460), solid culture (Lowenstein-Jensen [LJ] agar and selective 7H11 [7H11S] agar), and fluorescent acid-fast bacilli (AFB) smear were performed on all 3 specimens from each patient. Eighty-one (10.2%) of 797 patients tested were diagnosed with pulmonary TB. The sensitivity of NAAT, BACTEC, LJ, 7H11S, and smear for the first specimen was 90%, 85%, 67%, 53%, and 58%, respectively, whereas the sensitivity for the series of 3 specimens was 90%, 95%, 74%, 74%, and 70%, respectively. Positive predictive value was 100% for all tests except AFB smear, which was 79%. The time to detect 75% of all TB cases was 4 days for NAAT and 21 days for liquid culture; other tests had a sensitivity of less than 75%. Identification and testing every first diagnostic specimen by NAAT has the potential to reduce the overall TAT for laboratory TB diagnosis by approximately 2 weeks. PMID- 15893904 TI - In vitro nonspecific mitogenic response of T-cell subsets in acute and chronic brucellosis. AB - The aim of the present study was to determine the cellular immune response on the course of brucellosis by investigating the proliferation response of T-cell subsets to phytohemagglutinin (PHA), that is, nonspecific mitogen in the patients with acute and chronic brucellosis. The study was performed in 19 patients with untreated brucellosis (acute, n = 11; chronic, n = 8) and 19 healthy controls. Standard tube agglutination and Coombs tests for brucellosis were performed. CD4+ and CD8+ T cell were investigated by the flow cytometry and sorting methods in all of cases. After these cells were cultured and stimulated with PHA, [H3] thymidine uptake and stimulation indices (SIs) were established. In all of the patients with brucellosis, CD4+ SIs and CD8+ SIs were found to be 1.40 +/- 0.63 and 1.45 +/- 0.42, respectively, and in the controls CD4+ SIs and CD8+ SIs were 1.59 +/- 0.36 and 1.64 +/- 0.37, respectively. In acute cases, CD4+ SIs were 1.71 +/- 0.64 and CD8+ SIs were 1.54 +/- 0.45. CD4+ SIs were 0.97 +/- 0.25 and CD8+ SIs were 1.32 +/- 0.37 in chronic cases. Although in acute cases CD4+ SIs and CD8+ SIs were not different from those in the control group, CD4+ SIs of chronic brucellosis cases were found to be significantly low as compared with those of acute brucellosis cases and the controls (P < 0.01). CD8+ SIs of acute and chronic brucellosis cases were not found to be different from those in the controls. Brucella agglutination titers of the patients with acute and chronic brucellosis were not found related with CD4 SIs and CD8 SIs. The findings of significantly low results of CD4+ T-cell proliferative responses of chronic brucellosis to PHA as compared with control and acute brucellosis cases remind that the development of chronic infection might be a result of T-helper proliferation defect. PMID- 15893905 TI - The oncologic surgery of Serefeddin Sabuncuoglu in the 15th century. AB - Serefeddin Sabuncuoglu (AD 1385-1468) was the author of the first illustrated surgical textbook in the Turkish-Islamic literature, Cerrahiyyet'ul-Haniyye (Imperial Surgery), written in old Turkish in 1465. Cerrahiyyet'ul-Haniyye, consists of three chapters and 193 sections which include coloured miniatures of surgical procedures, incisions, and instruments, for a variety of surgical specialties, including oncology. Serefeddin Sabuncuoglu was a surgeon, medical teacher, deontologist and a miniature artist who should be remembered as a contributor to the modern medical practice six centuries later. PMID- 15893906 TI - The quality of chemotherapy and its quality assurance. AB - AIMS: Assessment of the quality of chemotherapy care and its quality assurance in clinical trials and daily practice. METHODS: Using Medline, literature was searched combining the following words: quality assurance or quality of care, combined with anti-neoplastic agents. The bibliography of each article was reviewed for additional literature. Those reports in English, French, German or Dutch focusing quality assurance or quality of care and chemotherapy were selected. RESULTS: One hundred and five articles were selected by Medline and after review and adding of additional literature 53 articles remained. In clinical trials information on quality of chemotherapy is sparse. Different cooperative groups reported on suboptimal dosing, suboptimal registration of chemotherapy and several trials indicated that suboptimal dosing led to impaired outcome. Most quality assurance activities in clinical trials are concerned with audit and feedback and on-site visits. In daily practice the quality of chemotherapy is mostly impaired by the fact that it is not given although indicated and if it is given non-evidence based chemotherapy or administration schedules and reduced dose intensity decrease the quality of care. Especially, age, comorbidity and socio-economic status reduce the chance of receiving good quality of care regarding chemotherapy. Activities mostly used for quality assurance are generation of guidelines, specialisation and multidisciplinary care. CONCLUSIONS: Most quality assurance activities in clinical trials and daily practice are directed to structure and process parameters. More evidence that quality of care is related to outcome should be sought. Quality assurance in daily practice should aim at guideline implementation, specialisation and multidisciplinary care and should pay attention especially to the older patients, patients with comorbidity and patients from lower socio-economic classes. PMID- 15893907 TI - Sentinel node biopsy or limited oriented axillary dissection (LOAD) in early breast cancer. AB - AIMS: The value of the sentinel node biopsy technique is recognised by the majority of surgical teams as an alternative to conventional axillary lymph node dissection for the treatment of small breast cancers. Secondary procedures are necessary when lymph node invasion not detected by frozen section examination is discovered post-operatively. In order to avoid or limit these reoperations, our sentinel node biopsy technique has gradually been transformed into limited oriented axillary dissection (LOAD), which avoids secondary procedures in the majority of patients. PATIENTS AND METHODS: Three hundred and eighty two patients were operated on by the same surgeon, using the patent blue sentinel node identification technique. This technique failed in nine patients, seven of whom were obese. Only one lymph node was removed in 75 patients, two in 88 patients, 3 5 in 174 patients and more than five lymph nodes were removed in the remaining patients. Eighty-eight percent of patients had no lymph node invasion on intraoperative and post-operative examination. RESULTS: Only seven patients were reoperated by secondary conventional lymph node dissection and there was residual cancer in only one patient. CONCLUSION: The oriented limited axillary dissection technique, combined with frozen section histological examination, avoids the usually unnecessary secondary operations in small breast cancers, in which axillary lymph node invasion rarely exceeds more than two nodes. This technique requires surgeons experienced in axillary surgery and conventional sentinel lymph node biopsy. It needs to be validated on a larger scale by a multicentre randomized prospective trial comparing LOAD to conventional axillary lymph node dissection. PMID- 15893908 TI - (18)F-FDG PET/CT in the evaluation of recurrent ovarian cancer: a prospective study on forty-one patients. AB - AIM: Many patients with ovarian cancer are at high risk of recurrence especially in the 2 years following first-line therapy. CA125 serum levels measurement associated to computed tomography (CT), ultrasound (US) and magnetic resonance imaging (MRI) are currently used during follow-up to detect recurrent disease. Unfortunately, in a relevant percentage of cases all of these traditional imaging techniques provide a significant number of doubtful/equivocal results or turn out negative even in presence of elevated Ca125 levels. Aim of our study was to evaluate sensitivity, specificity and accuracy of (18)F-FDG PET/CT in a group of patients with suspicion of ovarian cancer recurrence. METHODS: We prospectively evaluated 41 patients with a mean age of 59.4 years who had been previously treated for ovarian cancer with surgery and radio-chemotherapy or radio chemotherapy alone. Following the performance of traditional radiologic imaging (US, CT, MRI) and Ca125 measurement, all patients underwent additional (18)F-FDG PET/CT. PET/CT results were compared with histologic findings or clinical, laboratory and repeated traditional imaging techniques during subsequent follow up data. RESULTS: Of 41 patients 32 had a positive PET-CT (30 true positive, two false positive) whereas nine a negative PET/CT (five true negative, four false negative). Overall, in our experience (18)F-FDG PET/CT provided a good sensitivity (88.2%), specificity (71.4%) and accuracy (85.4%), superior to that reported in literature for traditional radiologic imaging. CONCLUSIONS: It can be concluded that (18)F-FDG PET/CT appears to be a useful and accurate tool in disclosing early recurrent ovarian cancer. PMID- 15893909 TI - The influence on treatment outcome of structuring rectal cancer care. AB - Clinical trials and registers data for quality assurance have been mandatory to achieve the good results and the enormous evolution which has been involved in rectal cancer surgery during the past 20 years. The whole business came into focus when local recurrences were considered as a matter of tumour biology and radiotherapy was introduced in many countries as a standard treatment in rectal cancer patients to reduce the local recurrence rate and to improve survival. During the last 30 years more than 8000 patients have been randomized in trials using pre- or post-operative radiotherapy. Those data are summarized in two good meta-analyses. In short, a summary of those meta-analyses has shown that radiotherapy reduces the local recurrence rate with 50%. Moreover, it has been revealed that pre-operative radiotherapy is better than post-operative radiotherapy in attempt to reduce the local recurrence rate and finally that there is a survival benefit with this reduction of the local recurrence rate. PMID- 15893910 TI - Prenatal exposure to the Dutch famine and disease in later life: an overview. AB - Low birth weight is associated with cardiovascular disease in adulthood. Poor maternal nutrition during gestation contributes to low birth weight. In this paper, we review the findings from a cohort of 2414 people, aged 50 years, born as term singletons around the time of the 1944-1945 Dutch famine, of which 912 people participated in an interview and 741 subjects were also available for hospital examination. We found more coronary heart disease, raised lipids, altered clotting and more obesity after exposure to famine in early gestation compared to those not exposed to the famine. Exposure in mid gestation was associated with obstructive airways disease and microalbuminuria. We found decreased glucose tolerance in people exposed to famine in late gestation. These findings show that maternal undernutrition during gestation has important effects on health in later life, but that the timing of the nutritional insult determines which organ system is affected. Future research should shed more light upon the underlying pathophysiology of the far-reaching effects of prenatal exposure to famine. PMID- 15893911 TI - Multimodal and three-dimensional imaging of prostate cancer. AB - Accurate characterization of prostate cancer is crucial for treatment planning and patient management. Non-invasive SPECT imaging using a radiolabeled monoclonal antibody, 111In-labeled capromab pendetide, offers advantage over existing means for prostate cancer diagnosis and staging. However, there are difficulties associated with the interpretation of these SPECT images. In this study, we developed a 3D surface-volume hybrid rendering method that utilizes multi-modality image data to facilitate diagnosis of prostate cancer. SPECT and CT or MRI (or both) images were aligned either manually or automatically. 3D hybrid rendering was implemented to blend prostate tumor distribution from SPECT in pelvis with anatomic structures from CT/MRI. Feature extraction technique was also implemented within the hybrid rendering for tumor uptake enhancement. Autoradiographic imaging and histological evaluation were performed to correlate with the in-vivo SPECT images. Warping registration of histological sections was carried out to compensate the deformation of histology slices during fixation to help the alignment between histology and in-vivo images. Overall, the rendered volumetric evaluation of prostate cancer has the potential to greatly increase the confidence in the reading of radiolabeled monoclonal antibody scans, especially in patients where there is a high suspicion of prostate tumor metastasis. PMID- 15893912 TI - Adaptive penalty likelihood for reconstruction of multidimensional confocal microscopy images. AB - In this paper we devise a penalty likelihood with noise constraints method to restore 2D and 3D confocal microscope images. Regularization is a commonly used technique in image restoration to balance restored image quality and noise suppression, but despite this noise is usually amplified. Taking into account common confocal imaging system degradation, we develop an algorithm by using a gradient descent method (PLGDA) to approach the minimum solution of the penalty likelihood equation. A Lagrange parameter controls the balance between the penalty and likelihood terms and is estimated using an adaptive method. We show that the a priori information is key to the regularization and Lagrange parameter estimation. The convergence characteristics are analysed and discussed. PLGDA and a traditional maximum likelihood expectation maximization are used to restore 2D and 3D confocal images. The point spread function (PSF), used to restore the data is collected from an experiment and modelled by bi-cubic splines to give an accurate noise free representation. Our experimental results show that the restored images are significantly improved by PLGDA. PMID- 15893913 TI - An integrated environment for plastic surgery support: building virtual patients, simulating interventions, and supporting intraoperative decisions. AB - In the last decade a number of environments for Computer Supported Plastic Surgery have been presented. Nevertheless, an overall approach for training and intraoperative support is still missing or has not been widely exploited yet. We developed a fully integrated system which allows surgical simulation, planning, and support for computer-guided plastic surgery procedures starting from image acquisition to final intraoperative assistance. The system also provides the user with a radiological workstation able to analyse patient medical images and case studies, with advanced bidimensional and three dimensional image processing functionalities. We intend to demonstrate that such a platform can be built at an affordable cost. The radiological workstation is capable of supporting radiologists and surgeons in real patient case studies and the simulation workstation may be adopted by plastic surgeons in teaching and training of complex surgical planning. Moreover, results of simulation can be used in the operating room with a relatively high benefit in terms of improved accuracy, reduction of surgical risks, and decrease in training costs. PMID- 15893914 TI - Study of myocardial glucose metabolism in rats with PET using wavelet analysis techniques. AB - The parametric images are attractive in PET imaging since they provide a global view of the imaged tissue including healthy and diseased structures. However these images are hampered by the noise due to the limited injected dose of the tracer. In this work we compare myocardial metabolic rates of glucose (MMRG) in rats obtained in the spatial and in the wavelet domains with the usual graphical method. The images were decomposed in the wavelet domain and all correlated pixels in approximations and details images were considered. Mean MMRG obtained from regions-of-interest drawn on parametric images in both methods showed values 15% higher while the variance was about 18% lower in the wavelet images. In conclusion, the wavelet filtering process allowed to enhance the image quality and to reduce the variance in the parametric images while preserving the spatial resolution. PMID- 15893915 TI - Oligomerization of the alpha and beta isoforms of the thromboxane A2 receptor: relevance to receptor signaling and endocytosis. AB - Thromboxane A(2) (TXA(2)) is a potent mediator of inflammation, vasoconstriction and oxidative stress. The TXA(2) receptor (TP) is a G protein-coupled receptor (GPCR) that is expressed as two alternatively spliced isoforms, alpha (343 residues) and beta (407 residues) that share the first 328 residues. For many years GPCRs were assumed to exist and function as monomeric species, but increasing evidence suggests that a dimer is the minimal functional unit of GPCRs. In the present report, using co-immunoprecipitation of differentially tagged TP expressed in HEK293 cells, we demonstrate that TPalpha and TPbeta form homo- and hetero-oligomers. Immunoblotting of lysates from human platelets with an anti-TP specific antibody revealed the presence of endogenously expressed TP oligomers. We show that TP oligomerization is an agonist-independent process highly affected by the reducing agent dithiothreitol suggesting the involvement of disulfide bonds in TP oligomerization. Over-expression of G protein-coupled receptor kinases and arrestins did not modulate the extent of receptor dimerization/oligomerization. Co-expression of two TP signaling-deficient mutants, R60L and E2402R, resulted in rescuing of receptor signal transduction suggesting that dimers/oligomers constitute the functional units of this receptor. Interestingly, TPalpha which does not undergo constitutive or agonist induced endocytosis on its own was subjected to both types of endocytosis when co expressed with TPbeta, indicating that TPalpha can display intracellular trafficking when complexed through hetero-oligomerization with TPbeta. PMID- 15893916 TI - Dissociation between medial temporal lobe and basal ganglia memory systems in schizophrenia. AB - The purpose of this study was to investigate basal ganglia (BG) and medial temporal lobe (MTL) dependent learning in patients with schizophrenia. Acquired equivalence is a phenomenon in which prior training to treat two stimuli as equivalent (if two stimuli are associated with the same response) increases generalization between them. The learning of stimulus-response pairs is related to the BG, whereas the MTL system participates in stimulus generalization. Forty three patients with DSM-IV schizophrenia and 28 matched healthy controls participated. Volunteers received the Rutgers acquired equivalence task (face fish task) by [Myers, C.E., Shohamy, D., Gluck, M.A. et al., 2003. Dissociating hippocampal versus basal ganglia contributions to learning and transfer. J. Cogn. Neurosci. 15, 185-193.], the California Verbal Learning Test (CVLT), and the n back working memory test. The Rutgers acquired equivalence task investigates BG dependent processes (stimulus-response learning) and MTL-dependent processes (stimulus generalization) with a single test. Results revealed that patients with schizophrenia showed a selective deficit on stimulus generalization, whereas stimulus-response learning was spared. The stimulus generalization deficit correlated with the CVLT performance (total scores from trials 1-5 and long-delay recall), but not with the n-back test performance. The number of errors during stimulus-response learning correlated with the daily chlorpromazine-equivalent dose of antipsychotics. In conclusion, this is the first study to show that patients with schizophrenia exhibit deficits during MTL-dependent learning, but not during BG-dependent learning within a single task. High-dose first generation antipsychotics may disrupt BG-dependent learning by blocking dopaminergic neurotransmission in the nigro-stiratal system. PMID- 15893917 TI - Quinolone antimicrobial resistance in some enterobacteria: a 10-year study in a Venezuelan general hospital. PMID- 15893918 TI - Uptake of fluorescein isothiocyanate-labelled dextran into the CSF after intranasal and intravenous administration to rats. AB - With the growing number of patients suffering from central nervous system (CNS) diseases a suitable approach for drug targeting to the brain becomes more and more important. In the present study, the contribution of the nose-CSF pathway to the uptake of the model drug fluorescein isothiocyanate-labelled dextran with a molecular weight of 3.0 kDa (FD3) into the CSF was determined in rats. FD3 was administered intranasally (489 microg/rat) and by intravenous infusion (24.4 microg/ml; 119 microg/rat) in the same set of animals (n=6). Blood samples were taken from the tail vein and CSF was sampled by cisternal puncture using a stereotaxic frame. The contribution of the olfactory pathway to the uptake of FD3 into the CSF was determined by comparing the AUCCSF/AUCplasma ratios after intranasal and after intravenous application of FD3 mimicking the blood levels after intranasal delivery. No significant difference was observed between the AUCCSF/AUCplasma ratios of FD3 after intranasal administration (1.33+/-0.40%) and intravenous infusion (1.03+/-0.56%). This indicates that in rats about 1% of the amount of FD3 in plasma reaches the CSF both after nasal and intravenous administration and that no direct transport of FD3 from the nose-CSF could be found. PMID- 15893919 TI - Controlled-release matrix tablets of ibuprofen using cellulose ethers and carrageenans: effect of formulation factors on dissolution rates. AB - The study was conducted to investigate the effects of carrageenans, and cellulose ethers on the drug release rates of ibuprofen controlled-release tablet matrices prepared by direct compression. Polymer blends containing carrageenans or cellulose ethers were used for the formulation and the effect of varying the polymer concentration on the release of the drug was studied. Other factors such as changes in surface topography of the matrices due to hydration were observed using a cryogenic scanning electron microscopy technique. Multiple regression analysis was used to predict the time for 50% release (t50) as a function of the concentration of the polymers used. Most of the formulations showed linear release profiles (r(2)>or=0.96-0.99) and sustained the release of ibuprofen over 12-16 h. The highest t50 (9.3 h) was for the formulation that contained a blend of 1:2 ratio of Viscarin and HPMC, while the lowest (3 h) was for the matrices that contained a 2:1 ratio of methylcellulose and Gelcarin. The majority of the matrix tablets that contained 10% polymer disintegrated prematurely. Of all the polymer blends that were investigated, the combination of Viscarin and HPMC gave almost linear release profiles over the entire range of concentration that was studied. The least effective combination was methylcellulose in combination with HPMC. Most of the formulations released ibuprofen by an anomalous (non-Fickian) transport mechanism, except those matrices that contained methylcellulose and Gelcarin (in a 1:1 and 1:2 ratio), which showed zero-order release. PMID- 15893920 TI - Simulation of fasting gastric conditions and its importance for the in vivo dissolution of lipophilic compounds. AB - In this study, the importance of accurate simulation of fasting gastric environment for the assessment of the absorption process of two model lipophilic compounds, GR253035X (weak base) and atovaquone (non-ionizable), was assessed. Dissolution profiles were constructed in previously proposed simulated gastric fluids and in a new medium that comprises only of components that have been recovered from the fasting stomach. Dissolution data obtained in a more physiologically relevant medium led to better correlation between the simulated and actual intralumenal dissolution vs. time profiles for GR253035X. In contrast, accurate simulation of gastric environment did not affect the simulated plasma profile of atovaquone. Accurate simulation of the fasting gastric contents may be crucial for the assessment of the absorption profile of lipophilic weak bases. PMID- 15893921 TI - Depression, anxiety and health status after hospitalisation for COPD: a multicentre study in the Nordic countries. AB - Patients with chronic obstructive pulmonary disease (COPD) often report anxiety, depression and poor health status, not least if they experience repeated hospitalisations due to acute exacerbations. The aim of this study was to analyse the interrelationships between health status, anxiety, depression and physical status in COPD patients being discharged after hospitalisation. This was a prospective study of 416 patients in five university hospitals in each of the Nordic countries. Data included demographic information, lung function and co morbidity. The Hospital Anxiety and Depression Scale and St. George's Respiratory Questionnaire (SGRQ) were applied to all patients. Both anxiety and depression were common among these patients. Anxiety was more common in women than in men (47% vs. 34%, P=0.009) and current smokers had a higher prevalence of both anxiety (54% vs. 37%) and depression (43% vs. 23%) than non-smokers (P<0.01). In general, the studied COPD patients had poor health status, especially those with anxiety, depression or both. Psychological status was independently related to all dimensions of SGRQ. Higher GOLD stages were significantly associated with increasing impairment in health status. In conclusion this multicentre study showed that anxiety and depression are common in patients with COPD, and, furthermore, that patients with psychological disorders have poor health status. Screening for depression and anxiety may help to identify patients with poor quality of life and an urgent need for intervention in order to improve their health status. PMID- 15893922 TI - Bronchoesophageal fistula due to broncholithiasis: a case series. AB - OBJECTIVE: To describe the clinical presentation, diagnostic evaluation, and treatment of bronchoesophageal (BE) fistula due to broncholithiasis, a rare cause of chronic cough with specific therapeutic implications. METHODS: Retrospective chart review of 9 patients diagnosed with BE fistulae at our tertiary-care institution between 1964 and 2002. RESULTS: The median age of patients (3 men, 6 women) was 56 years (range, 34-72 years). Six patients had never smoked. Eight presented with intractable cough typically worse after drinking that was relieved by lying on either side; 1 patient presented with dysphagia. Six patients reported lithoptysis, and most had a history of recurrent pneumonia. Chest findings were nonspecific. Diagnosis was established by radiocontrast studies of the esophagus (5 patients), surgical procedures (3), or bronchoscopy (1). Fistulae were right-sided in 7 patients and left-sided in 2. All patients had surgical repair of the fistulae; 6 patients experienced symptom resolution. CONCLUSION: BE fistula caused by broncholithiasis most commonly affects the right bronchial tree and should be considered in patients with chronic cough associated with drinking, lithoptysis, or recurrent pneumonia. The diagnosis is usually established by radiocontrast studies of the esophagus or incidentally during operations. Surgical repair is required. PMID- 15893923 TI - Global Initiative on Obstructive Lung Disease (GOLD) classification of lung disease and mortality: findings from the Atherosclerosis Risk in Communities (ARIC) study. AB - OBJECTIVE: To determine whether a modified Global Initiative on Obstructive Lung Diseases (GOLD) classification of chronic obstructive pulmonary disease (COPD) predicts mortality in a cohort of subjects followed for up to 11 years. METHODS: We analyzed data from 15,759 adult participants, aged 43-66 years at baseline, in the Atherosclerosis Risk in Communities (ARIC) study. All baseline and follow-up data were available for 15,440 (97.9%) of the initial participants. We classified subjects using a modification of the GOLD criteria for COPD (prebronchodilator forced expiratory volume in 1s (FEV(1)) stratification of disease severity), and added a "restricted" category (FEV(1)/FVC>70% and FVC<80% predicted). We used Cox proportional hazard models to determine the risk of impaired lung function on subsequent mortality, after adjusting for age, race, sex and smoking status. RESULTS: 1242 (8.0%) subjects died by the end of 1997. The overall rate of death was 8.9 per 1000 person years, but varied from 5.4/1000 among normal subjects to 42.9/1000 among subjects with GOLD Stage 3 or 4 COPD. After adjusting for covariates, all GOLD categories, along with the restricted category, predicted a higher risk of death: GOLD Stage 3 or 4, hazard ratio (HR) 5.7, 95% confidence interval (CI) 4.4, 7.3; GOLD Stage 2 HR 2.4, 95% CI 2.0, 2.9; GOLD Stage 1 HR 1.4, 95% CI 1.1, 1.6; GOLD Stage 0 HR 1.5, 95% CI 1.3, 1.8; and restricted HR 2.3, 95% CI 1.9, 2.8. CONCLUSION: The modified GOLD classification system of COPD predicts mortality in this cohort of middle-aged Americans followed for up to 11 years. PMID- 15893924 TI - Screen-printed bienzymatic sensor based on sol-gel immobilized Nippostrongylusbrasiliensis acetylcholinesterase and a cytochrome P450 BM-3 (CYP102-A1) mutant. AB - Here, we describe the development of a bi-enzymatic biosensor that simplifies the sample pretreatment steps for insecticide detection, and opens the way for a highly sensitive detection of phosphorothionates in food. These compounds evolve their inhibitory activity towards acetylcholinesterases (AChEs) only after oxidation, which is performed in vivo by P450 monooxygenases. Consequently, phosphorothionates require a suitable sample pretreatment by selective oxidation to be detectable in AChE based systems. In this study, enzymatic phosphorothionate activation and AChE inhibition were integrated in a single biosensor unit. A triple mutant of cytochrome P450 BM-3 (CYP 102-A1) and Nippostrongylus brasiliensis AChE (NbAChE) was immobilized using a fluoride catalyzed sol-gel process. Different sol-gel types were fabricated and characterized regarding enzyme loading capacity and enzyme activity containment. The enzyme sol-gel itself already proved to be suitable for the highly sensitive detection of paraoxon and parathion in a spectrometric assay. A method for screen printing of this enzyme sol-gel on thick film electrodes was developed. Finally, amperometric biosensors containing coimmobilized NbAChE and the cytochrome P450 BM-3 mutant were produced and characterized with respect to signal stability, organophosphate detection, and storage stability. The detection limits achieved were 1 microg/L for paraoxon and 10 microg/L for parathion, which is according to EC regulations the highest tolerable pesticide concentration in infant food. PMID- 15893925 TI - Piezoelectric immunochip for the detection of dengue fever in viremia phase. AB - The global prevalence of dengue fever has grown so dramatically in recent years that it is endemic in more than 100 countries and has become a major international public health concern. Moreover, since the flu-like symptoms that accompany dengue fever are atypical and varied, the detection procedures currently used to identify it are cumbersome and time-consuming, making early stage epidemiological control and effective medical treatment of this epidemic almost impossible. In this study, a QCM-based detection system was developed in which two monoclonal antibodies against dengue E and NS-1 protein, respectively, were control orientated immobilized on QCM via protein A to produce an immunochip. Various sample pretreatment procedures were evaluated to ascertain the most suitable combination, and both the simulating samples and the clinical specimen were examined by the immunochip. The results revealed that the cibacron blue 3GA gel-heat denature (CB-HD) method was the most effective sample pretreatment technique. Due to the complex composition of the serum, the immunochip could only effectively quantify dengue viral antigens in a 1/1000 untreated simulated sample. With the help of the CB-HD method, the dilution folds were found to capable of being reduced from 1000 to 100, and the detection limit lowered to 1.727 microg/ml (E protein) and 0.740 microg/ml (NS-1 protein) in the original sample. While the cocktail immunochip could not quantify both antigens separately, the higher signal level rendered it a more effective qualification tool for suspect screening. Moreover, the results of the analysis of clinical specimens also proved the ability and future potential of cocktail immunochip in discriminating dengue-positive cases from negative serum specimens in the viremia phase. PMID- 15893926 TI - Molecular cloning of P450 aromatase from the leopard gecko and its expression in the ovary. AB - In this study, we identified the cDNA of P450 aromatase in the leopard gecko, a lizard with temperature-dependent sex determination. The cDNA encodes a putative protein of 505 amino acids. The deduced amino acid sequence of leopard gecko aromatase cDNA showed 80% identity with that of turtles, 70% with humans and 77% with chickens. This is the first report of the identification of P450 aromatase cDNA in squamata species. It has been reported that this gene is expressed in different layers of cells in the ovary of mammalian species and avian species. Thus, we also investigated cells expressing the mRNA of this gene in the ovary of the leopard gecko by RT-PCR and in situ hybridization. The mRNA expression of leopard gecko P450 aromatase was localized in both the thecal and granulosa cell layers in the ovary. The expression in thecal and granulosa cell layers was examined in the largest follicle, second largest follicle and third largest follicle by RT-PCR. A higher level of mRNA expression was observed in the granulosa cell layer of the second largest follicle than in other cell layers. This result may reflect the characteristics of follicles in species with automonochronic ovulation. PMID- 15893927 TI - Hologram quantitative structure-activity relationships for a series of farnesoid X receptor activators. AB - The farnesoid X receptor (FXR) is an attractive drug target for the development of novel therapeutic agents for the treatment of dyslipidemia and cholestasis. Hologram quantitative structure-activity relationship (HQSAR) studies were conducted on a series of potent FXR activators originated from natural product like libraries. A training set containing 82 compounds served to establish the models. The best HQSAR model was generated using atoms, bonds, connections, chirality, and donor and acceptor as fragment distinction and fragment size default (4-7) with six components. The model was used to predict the potency of 20 test set compounds that were not included in the training set, and the predicted values were in good agreement with the experimental results. The final HQSAR model and the information obtained from HQSAR 2D contribution maps should be useful for the design of novel FXR ligands having improved potency. PMID- 15893928 TI - Synthesis and biological evaluation of chalcones and their derived pyrazoles as potential cytotoxic agents. AB - A series of substituted chalcones and their corresponding pyrazoles were synthesized and evaluated for in vitro cytotoxic activity against a panel of human cancer cell lines. Out of 93 compounds screened, 8 compounds, 1s, 3i,j,n, 4i,j,n and 4s, showed marked activity. Compounds 4j,n and 4s were found to be the most promising in this study. SAR is also discussed. PMID- 15893929 TI - Beta-lactam compounds as apparently uncompetitive inhibitors of HIV-1 protease. AB - Compounds of a combinatorial monocyclic beta-lactam library were found to be apparently uncompetitive inhibitors of HIV-1 protease, providing lead compounds for a new class of HIV protease inhibitors. PMID- 15893930 TI - Novel semicarbazide-derived inhibitors of human dipeptidyl peptidase I (hDPPI). AB - Human dipeptidyl peptidase I (hDPPI, cathepsin C, EC 3.4.14.1) is a novel putative drug target for the treatment of inflammatory diseases. Using 1 as a starting point (IC50>10 microM), we have improved potency by more than 500-fold and successfully identified novel inhibitors of DPPI via screening of a one-bead two-compounds library of semicarbazide derivatives. Selected compounds were shown to inhibit intracellular DPPI in RBL-2H3 cells. These compounds were further characterized for adverse effects on HepG2 cells (cytotoxicity and viability) and their metabolic stability in rat liver microsomes was estimated. One of the most potent inhibitors, 8 (IC50=31+/-3 nM; Ki=45+/-2 nM, competitive inhibition), is selective for DPPI over other cysteine and serine proteases, has a half-life of 24 min in rat liver microsomes, shows approximately 50% inhibition of intracellular DPPI at 20 microM and is noncytotoxic. PMID- 15893931 TI - Synthesis of a C-terminally biotinylated macrocyclic peptide mimetic exhibiting high Grb2 SH2 domain-binding affinity. AB - Although considerable effort has been devoted to developing Grb2 SH2 domain binding antagonists, important questions related to ligand specificity, and identification of intracellular targets remain unanswered. In order to begin addressing these issues, the design, synthesis, and evaluation of a novel biotinylated macrocycle are reported that bears biotin functionality at a C terminal rather than the traditional N-terminal position. With a Grb2 SH2 domain binding K(eq) value of 3.4 nM, the title macrocycle (5) is among the most potent biotinylated SH2 domain-binding ligands yet disclosed. This should be a useful tool for elucidating physiological targets of certain Grb2 SH2 domain-binding antagonists. PMID- 15893932 TI - Radical scavenging activity of dicaffeoyloxycyclohexanes: contribution of an intramolecular interaction of two caffeoyl residues. AB - Six regio- and stereoisomers of dicaffeoyloxycyclohexanes and 2,4-di-O-caffeoyl 1,6-anhydro-beta-D-glucose were synthesized as model compounds of dicaffeoylquinic acids, and their radical scavenging activity was evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3 ethylbenzthiazoline-6-sulfonic acid) diammonium salt) radical scavenging tests. Both DPPH and ABTS radical scavenging reactions of these compounds consisted of two different steps. In the first step, catechol moieties of the caffeoyl residues were rapidly converted to o-quinone structures and no significant difference in the reactivity was observed among the tested compounds. In the second step, however, the rate of the reaction increased as the intramolecular distance of the two caffeoyl residues decreased. A novel intramolecular coupling product, which could scavenge additional radicals, was isolated from the reaction mixture of trans-1,2-dicaffeoyloxycyclohexane and DPPH radical. The result suggests that the second step of the radical scavenging reaction is arising from an intramolecular interaction between the two caffeoquinone residues to regenerate catechol structures, and that the closer their distance is, the more rapidly they react. The radical scavenging activity of natural dicaffeoylquinic acids in a biological aqueous system might also depend on the positions of caffeoyl ester groups. PMID- 15893933 TI - Determination of human plasma phospholipid transfer protein mass and activity. AB - Plasma phospholipid transfer protein (PLTP) plays an important role in lipoprotein metabolism. PLTP is an 80-kDa glycoprotein that is expressed/secreted by a wide variety of tissues including lung, liver, adipose tissue, brain, and muscle. PLTP mediates a net transfer of phospholipids between vesicles and plasma HDLs. It also generates from small HDL particles large fused HDL particles with a concomitant formation of small lipid-poor apolipoprotein (apo) A-I-containing particles which are thought to act as the primary acceptors of cell-derived cholesterol from peripheral tissue macrophages. Another important function of PLTP is connected to lipolysis. Its role in the transfer of surface remnants from triglyceride-rich particles, very-low-density lipoproteins, and chylomicrons, to HDL is of importance for the maintenance of HDL levels. Recent observations from our laboratory have demonstrated that in circulation two forms of PLTP are present, one catalytically active (high-activity form, HA-PLTP) and the other a low-activity form (LA-PLTP). In view of the likely relevancy of PLTP in human health and disease, reliable and accurate methods for measuring plasma/serum PLTP activity and concentration are required. In this chapter, two radiometric PLTP activity assays are described: (i) exogenous, lipoprotein-independent phospholipid transfer assay and (ii) endogenous, lipoprotein-dependent phospholipid transfer assay. In addition, an ELISA method for quantitation of serum/plasma total PLTP mass as well as HA-PLTP and LA-PLTP mass is reported in detail. PMID- 15893934 TI - Development of experimental designs for atherosclerosis studies in mice. AB - The mouse has become the de facto model for the majority of atherosclerosis studies. Studies involving the quantification of lesions in mouse models of the disease represent the basis of our evolving concepts on the biochemical and cellular mechanisms underlying the atherogenic process. Many issues of experimental design, including specific model, strain, gender, atherogenic stimulus, duration of study, group size, and statistical analysis may influence the outcome and interpretation of atherosclerosis studies. The selection of vascular bed in which to quantify atherosclerotic lesion size could also impact the interpretation of results. Early studies quantified atherosclerotic lesion size in either specific regions or all of the aortic sinus. Measurement of atherosclerosis throughout the aortic intimal surface has become a common mode for defining lesion size. It is likely that other vascular regions will be increasingly used. In addition to size, there is an increased emphasis on identifying and quantifying the cellular and chemical composition of atherosclerotic lesions. PMID- 15893935 TI - Studying phospholipid metabolism using yeast systematic and chemical genetics. AB - Most phospholipid metabolic pathways in the budding yeast Saccharomyces cerevisiae are analogous to their mammalian counterparts. The biological tractability of yeast provides for an opportunity to rapidly determine functions of specific lipids or lipid metabolic pathways using both classical and chemical genetic techniques. The recent generation of the yeast genome deletion collection revealed that approximately 75% of yeast genes are not essential for life. Coupling analysis of the yeast deletion collection with automation using high throughput robotics enables yeast genetic screens to be more thorough and bypasses the requirement for library screens to identify genes of interest. Two high-throughput yeast genetic methods are described, systematic synthetic lethality and chemical genetics. Systematic synthetic lethality is based on the principle that inactivation of two genes separately has minimal effects on cell growth whereas inactivation of both genes simultaneously results in growth defects due to their shared requirement in a particular cellular process. Chemical genetics is the analysis of bioactive compounds to determine processes that regulate susceptibility to the compound under study, and provides powerful data regarding precise targets and mechanism of action that regulate action of the compound. PMID- 15893936 TI - Techniques to measure lipase and esterase activity in vitro. AB - Lipases and esterases constitute a large category of enzymes. They are ubiquitous in nature, found in bacteria, fungi, and animals. The family members address a wide variety of structurally diverse substrates. Appropriately, a large number of assays have been developed to analyze their activity in vitro. Here, we present an overview of these enzymes, along with protocols for common assays performed in solution. An emphasis is placed on assays for enzymes that can hydrolyze triacylglycerols. PMID- 15893937 TI - LDL receptor family: isolation, production, and ligand binding analysis. AB - Members of the low density lipoprotein receptor gene family have recently received particular attention because of their involvement not only in lipoprotein transport, but also in signal transduction pathways. The main characteristic feature of this protein group is their cysteine-rich ligand binding domain, which is able to bind many unrelated proteins, such as apolipoproteins, proteases, and protease/inhibitor complexes, signaling molecules such as reelin, and several other groups of proteins. The main challenges of studying these proteins in vitro are their extremely high content of disulfide bridges and the detergent-sensibility of their classical ligands, i.e, lipoproteins. Here, we describe generally applicable procedures for the analysis of these receptors. We present an outline of established methodology for their isolation and visualization, the production of recombinant fragments, in particular of soluble ligand binding domains, and we describe standard procedures for the analysis of the functionality of the receptors and recombinant receptor ligand binding fragments, respectively. PMID- 15893938 TI - Neuronal models for studying lipid metabolism and transport. AB - New methods have been developed for studying lipid metabolism and transport in primary cultures of neurons. Sympathetic neurons from rats and mice, as well as retinal ganglion neurons from rats, can be cultured in three-compartmented culture dishes in which the cell bodies reside in a compartment separate from that housing the distal axons. In addition, the three compartments contain completely independent fluid environments. Consequently, these neuronal cultures represent an excellent model for studying the intra-neuronal transport of lipids and proteins between cell bodies and distal axons. In addition, compartmented neuron cultures are particularly appropriate for investigating factors that regulate axonal growth and neuronal survival. The application of the compartmented culture model for use with murine neurons has opened up many new possibilities for studying lipid metabolism in neurons derived from genetically modified mice. Examples are given in which compartmented cultures of primary neurons have been used in studies on (i) lipid analysis of distal axons and cell bodies/proximal axons, (ii) immunoblotting of neuronal proteins involved in lipid metabolism, (iii) the compartmentalization of lipid metabolism, (iv) the role of lipids in axonal growth and survival, and (v) intracellular lipid transport. PMID- 15893939 TI - Expression and one-step ion-exchange purification of (AAR)IL-8 (human IL-8 receptor antagonist). AB - Interleukin-8 (IL-8) is C-X-C chemokine, which is produced by a variety of cells. IL-8 plays an important role in the inflammatory response and may be a therapeutic target for some inflammatory diseases. To develop an IL-8 receptor antagonist, (AAR)IL-8 (IL-8 receptor antagonist) was constructed and successfully expressed in Escherichia coli. (AAR)IL-8 could be easily purified by one-step SP Sepharose fast flow column after the lysate of recombinant bacterial cells was heated at 70 degrees C for 10 min. The purity of (AAR)IL-8 is more than 95%. This purification process resulted in final purified yields of 4.29 mg (AAR)IL-8/g cell paste. In addition, the purified (AAR)IL-8 can significantly inhibit the chemotaxis that was induced by human IL-8 in vitro and in vivo. These results showed that this purification process is very simple and effective. It could be easily amplified at a larger scale. (AAR)IL-8 might find use as a new therapeutic IL-8 receptor antagonist for some acute and chronic inflammatory diseases. PMID- 15893940 TI - Modulation of the semantic system by word imageability. AB - A prevailing neurobiological theory of semantic memory proposes that part of our knowledge about concrete, highly imageable concepts is stored in the form of sensory-motor representations. While this theory predicts differential activation of the semantic system by concrete and abstract words, previous functional imaging studies employing this contrast have provided relatively little supporting evidence. We acquired event-related functional magnetic resonance imaging (fMRI) data while participants performed a semantic similarity judgment task on a large number of concrete and abstract noun triads. Task difficulty was manipulated by varying the degree to which the words in the triad were similar in meaning. Concrete nouns, relative to abstract nouns, produced greater activation in a bilateral network of multimodal and heteromodal association areas, including ventral and medial temporal, posterior-inferior parietal, dorsal prefrontal, and posterior cingulate cortex. In contrast, abstract nouns produced greater activation almost exclusively in the left hemisphere in superior temporal and inferior frontal cortex. Increasing task difficulty modulated activation mainly in attention, working memory, and response monitoring systems, with almost no effect on areas that were modulated by imageability. These data provide critical support for the hypothesis that concrete, imageable concepts activate perceptually based representations not available to abstract concepts. In contrast, processing abstract concepts makes greater demands on left perisylvian phonological and lexical retrieval systems. The findings are compatible with dual coding theory and less consistent with single-code models of conceptual representation. The lack of overlap between imageability and task difficulty effects suggests that once the neural representation of a concept is activated, further maintenance and manipulation of that information in working memory does not further increase neural activation in the conceptual store. PMID- 15893941 TI - Two distinct neural effects of blinking on human visual processing. AB - Humans blink every few seconds, yet the changes in retinal illumination during a blink are rarely noticed, perhaps because visual sensitivity is suppressed. Furthermore, despite the loss of visual input, visual experience remains continuous across blinks. The neural mechanisms in humans underlying these two phenomena of blink suppression and visual continuity are unknown. We investigated the neural basis of these two complementary behavioural effects using functional magnetic resonance imaging to measure how voluntary blinking affected cortical responses to visual stimulation. Two factors were independently manipulated in a blocked design; the presence/absence of voluntary blinking, and the presence/absence of visual stimulation. To control for the simple loss of visual input caused by eyelid closure, we created a fifth condition where external darkenings were dynamically matched to each subjects' own blinks. Areas of lateral occipital cortex, including area V5/MT, showed suppression of responses to visual stimulation during blinking, consistent with the known loss in visual sensitivity. In contrast, a medial parieto-occipital region, homologous to macaque area V6A, showed responses to blinks that increased when visual stimulation was present. Our data are consistent with a role for this region in the active maintenance of visual continuity across blinks. Moreover, both suppression in lateral occipital and activation in medial parieto-occipital cortex were greater during blinks than during matched external darkenings of the visual scene, suggesting that they result from an extra-retinal signal associated with the blink motor command. Our findings therefore suggest two distinct neural correlates of blinking on human visual processing. PMID- 15893942 TI - fMRI-acoustic noise alters brain activation during working memory tasks. AB - Scanner noise during functional magnetic resonance imaging (fMRI) may interfere with brain function and change blood oxygenation level dependent (BOLD) signals, a problem that generally worsens at the higher field strengths. Therefore, we studied the effect of increased acoustic noise on fMRI during verbal working memory (WM) processing. The sound pressure level of scanner noise was increased by 12 dBA from "Quiet" to "Loud" echo planar imaging (EPI) scans by utilizing resonant vibration modes of the gradient coil. A WM paradigm with graded levels of task difficulty was used to further access WM load. Increased scanner noise produced increased BOLD responses (percent signal change) bilaterally in the cerebellum, inferior (IFG), medial (medFG), and superior (SFG) frontal, fusiform (FusG), and the lingual (LG) gyri, and decreased BOLD responses bilaterally in the anterior cingulate gyrus (ACG) and the putamen. This finding suggests greater recruitment of attention resources in these brain regions, probably to compensate for interference due to louder scanner noise. Increased working memory load increased the BOLD signals in IFG and the cerebellum, but decreased the BOLD signals in the putamen and the LG. These findings also support the idea that brain function requires additional attention resources under noisier conditions. Load- and acoustic-noise-related changes in BOLD responses correlated negatively in the WM network. This study demonstrates that MR noise affects brain activation pattern. Future comparisons between studies performed under different acoustic conditions (due to differing magnetic field strengths, pulse sequences, or scanner manufacturers) might require knowledge of the sound pressure level of acoustic noise during fMRI. PMID- 15893943 TI - Localization of the angiopoietin receptors Tie-1 and Tie-2 on the primary cilia in the female reproductive organs. AB - Blood vessel homeostasis and endothelial cell survival depend on proper signalling through angiopoietin receptors such as the receptor tyrosine kinases Tie-1 and Tie-2. We have studied the presence and subcellular localization of these receptors in murine female reproductive organs using confocal microscopy analysis of antibody stained tissue sections of ovary and oviduct. We show that Tie-2 principally localizes to primary cilia of the surface epithelium of the ovary, bursa and extra-ovarian rete ducts as well as to plasma membranes of ovarian theca and endothelial cells. Primary cilia of follicular granulosa cells were negative. Further, Tie-1 and Tie-2 localized to motile cilia of the oviduct. Western blotting detection and immunolocalization of anti-Tie-2 in ovary and oviduct were abolished by administration of an anti-Tie-2 blocking peptide, confirming antibody specificity. In a series of immunohistochemical analysis on human ovarian tissues we also observed a unique localization of Tie-2 to the primary cilia of ovarian surface epithelium. These observations are the first to show ciliary localization of angiopoietin receptors. Our results support the hypothesis that cilia of the female reproductive organs play a novel and important sensory role in relaying physiochemical changes from the extracellular environment to epithelial cells of the oviduct, the ovary and extra-ovarian tissues. PMID- 15893944 TI - Role of exosomes in sperm maturation during the transit along the male reproductive tract. AB - Even tough differentiated spermatozoa are unable of transcriptional or translational activity; the sperm surface undergoes major modifications in macromolecules composition during the transit along the male reproductive tract. This is the result of sequential, well orchestrated interactions between the male reproductive tract secretions and the transiting male gamete. This is particularly true when spermatozoa transit along the epididymis. The epididymis is a long convoluted tubules in which the spermatozoa leaving the testis have to transit. The unraveled epididymal tubule can be as long as 80 m in stallion, and the transit time of spermatozoa is of 3-12 days depending on the species. The epididymis is usually divided in three segments: the caput (proximal part), the corpus, and cauda. While the cauda epididymides acts as a sperm reservoir, the caput and corpus are responsible for sperm maturation. This means that, under androgen control, the epididymal epithelium secretes proteins that will interact sequentially with sperm surface. Some of the sperm proteins acquired during maturation along the excurrent duct behave as integral membrane proteins. In fact, some epididymal originating proteins are glycosylphosphatidylinositol (GPI) anchored to the sperm plasma membrane. Our laboratory has shown that some of these proteins are secreted in an apocrine manner by the epididymal epithelium and are associated to exosomes, called epididymosomes. Epididymosomes are rich in sphingomyelin and are characterized by a high cholesterol/phospholipids ratio. Many proteins are associated to epididymosomes, some of which are selectively transferred to spermatozoa during the epididymal transit. We have identified some of these exosomes associated proteins transferred to the maturing spermatozoa. These include two enzymes involved in the polyol pathway: an aldose reductase and a sorbitol dehydrogenase. A cytokine named MIF (macrophage migration inhibitory factor) is another protein associated to exosomes who is transferred to spermatozoa during the epididymal transit. We hypothesized that both the polyol pathway and MIF secreted in an apocrine fashion by the epididymal epithelium modulate sperm motility during the transit along the male reproductive tract. Finally, P25b, belonging to a family of sperm surface proteins (P26h/P34H) necessary for the binding to the surface of the egg, is also acquired through the interaction between epididymosomes and the male gamete. In vitro studies have defined the conditions of protein transfer when epididymal spermatozoa are co incubated with epididymosomes. The transfer of selected proteins to specific membrane domains of spermatozoa is saturable, temperature and pH-dependent, being optimal at pH 6.5. The presence of zinc in the incubation medium, but not of calcium neither magnesium, significantly increases the efficiency of protein transfer. These results show that exosomes play a role in sperm epididymal maturation which is an essential event to produce male gametes with optimal fertilizing ability. PMID- 15893945 TI - Inducible nitric oxide synthase expression is inhibited by myeloperoxidase. AB - Nitric oxide (NO) plays key roles in vasodilation and host defense, yet the overproduction of NO by inducible nitric oxide synthase (iNOS) at inflammatory sites can also be pathogenic. Here, we investigate the role of MPO in modulating the induction of iNOS by IFNgamma/LPS (IL). In monocyte-macrophages (Mvarphi) treated with IL, MPO gene expression was found to be downregulated as iNOS was upregulated. In Mvarphi from MPO-knockout (KO) mice, the induction of iNOS by IL was earlier and higher than in MPO-positive cells, suggesting MPO is inhibitory. Consistent with that interpretation, the addition of purified MPO enzyme to cultured macrophages inhibited iNOS induction by IL. In addition, an inhibitor of MPO enzyme, 4-aminobenzohydrazide, enhanced iNOS induction in MPO-positive cells, but not in MPO-KO cells. Similarly, taurine, a scavenger of MPO-generated HOCl, enhanced iNOS induction in MPO-positive cells, but not in MPO-KO cells. MPO affects an early event, suppressing iNOS induction when added within 2h of IL, but not when added several hours after IL. The suppression by MPO was alleviated by NO donor, sodium nitroprusside, suggesting the suppression results from scavenging of NO by MPO. This interpretation is consistent with earlier reports that MPO consumes NO, and that low levels of NO donor augment induction of iNOS by IFNgamma/LPS. The implication of these findings is that MPO acts as gatekeeper, suppressing the deleterious induction of iNOS at inflammatory sites by illegitimate signals. The combined signaling of IFNgamma/LPS overrides the gatekeeper function by suppressing MPO gene expression. PMID- 15893946 TI - Crossed-coil detection of two-photon excited nuclear quadrupole resonance. AB - Applying a recently developed theoretical framework for determining two-photon excitation Hamiltonians using average Hamiltonian theory, we calculate the excitation produced by half-resonant irradiation of the pure quadrupole resonance of a spin-3/2 system. This formalism provides expressions for the single-quantum and double-quantum mutation frequencies as well as the Bloch-Siegert shift. The dependence of the excitation strength on RF field orientation and the appearance of the free-induction signal along an axis perpendicular to the excitation field provide an unmistakable signature of two-photon excitation. We demonstrate single and double-quantum excitation in an axially symmetric system using 35Cl in a single crystal of potassium chlorate (omega(Q) = 28 MHz) with crossed-coil detection. A rotation plot verifies the orientation dependence of the two-photon excitation, and double-quantum coherences are observed directly with the application of a static external magnetic field. PMID- 15893947 TI - Comparative skin permeability of neonatal and adult timber rattlesnakes (Crotalus horridus). AB - Skin permeability and lipid content were determined using shed epidermis of neonatal and adult timber rattlesnakes (Crotalus horridus) from the Coastal Plain Pine Barrens of New Jersey and from the Appalachian Mountains of northern Pennsylvania. Differences between populations due to habitat and within populations due to age were tested. Skin permeability was not found to differ according to locality (P>0.05), but rates were significantly different for age. Permeability of adult epidermis was greater than that of neonates (P<0.01). Lipid content did not differ by locality (P>0.05), but differed between ages, paralleling the results found for permeation rates. Neonate sheds had a greater amount of extractable lipids than adult sheds (P<0.01). Despite the lower skin permeability of neonates, our estimates indicate that the percentage of their total body water content lost per hour may still be 2.2 times that of adults. Resistance to cutaneous water loss may be advantageous to neonates given their relatively large surface area-to-volume ratio. PMID- 15893948 TI - Red blood cell function and haematology in two tropical freshwater fishes from Australia. AB - Salmon catfish and tarpon occur in habitats that periodically become deficient in oxygen resulting in high mortalities of other fish species. The water-breathing catfish, Arius leptaspis, and the facultative air-breathing tarpon, Megalops cyprinoides, both have high haemoglobin and haematocrit, and the oxygen carrying capacity in the air-breather is exceptionally high (15.6+/-1.2 vol%). Iso-pH oxygen equilibria of the red blood cells at 25 degrees C revealed high affinity (P(50)=9 mmHg, pH 7.4) and co-operativity (n(50)>2.2, pH 7.4) in the catfish, and contrasted with low affinity (P(50)=32 mmHg, pH 7.4) and co-operativity (n(50) approximately 1) in the air-breathing tarpon. Oxygen binding was further distinguished by relative pH insensitivity (Bohr factor, O=Deltalog P(50)/Deltalog pH=-0.22) in the catfish, compared with a significant Bohr effect in the tarpon (O=-0.96). The potential for modulation of haemoglobin-oxygen affinity was indicated by a high ratio of GTP to ATP in the erythrocytes of the catfish, whereas regulation in the tarpon appeared due to ATP alone. Differences in blood respiratory functions between the two species are likely to reflect reduced opportunity for activity under extreme hypoxia in the catfish. PMID- 15893949 TI - Do arachidonic acid and its metabolites, secreted by rheumatoid and osteoarthritic synovial tissue, account for the strong inhibition of DNA synthesis in cultured human articular chondrocytes? A novel approach to the mechanism of tissue damage. AB - AIMS: To further characterize factors secreted in vitro by osteoarthritic and rheumatoid arthritis synovial membranes that inhibit DNA synthesis by cultured human articular chondrocytes, and extend these findings to synovial fluid. MATERIAL AND METHODS: Synovial tissue, synovial fluid and articular cartilage were obtained at surgery from two patients suffering rheumatoid arthritis and two other patients suffering from osteoarthritis. Synovial tissue was incubated in DMEM, then condition media and synovial fluids were extracted with methanol. Methanol extracts and extracted residues (hyaluronic acid, proteins) were assayed for their capacity to inhibit DNA synthesis in articular chondrocytes. Methanol extracts were also fractionated by thin layer chromatography on silica-coated plates and recovered fractions similarly tested. RESULTS: All extracts exhibited strong and concentration-dependent inhibition of [3H]-thymidine incorporation. The most potent inhibition was obtained with the extracts from rheumatoid joints and the least potent inhibition was with synovial fluids. The removal of active substances with methanol leaves an inactive residue. Methanol extraction does not alter the mitogenic activity of five exogenous growth factors and two cytokines, thus suggesting that such activity is entirely due to lipids. The bulk of anti mitotic factors extracted by methanol co-migrate when fractionated by thin layer chromatography on silica-coated plates with arachidonic acid and its lipo oxygenase metabolites. IN CONCLUSION: Inflamed synovium produces and releases lipids, most probably arachidonic acid metabolites that inhibit cell proliferation thus limiting inflammation and pannus formation in arthritis. PMID- 15893950 TI - Bone status in primary hyperparathyroidism assessed by regional bone mineral density from the whole body scan and QUS imaging at calcaneus. AB - To assess the bone mineral density status in primary hyperparathyroidism (PHPT), we studied 64 females with PHPT and 17 healthy women. Regional BMD (arms, trunk, legs) from the whole body scan and conventional sites (lumbar spine, femur, radius) were assessed by DXA. Quantitative ultrasound (QUS) imaging measurements were performed at calcaneus. Sixteen women had history of renal lithiasis, 11 had low impact fracture and 37 women had neither renal lithiasis nor fracture. In the entire group, the mean Z-scores were significantly decreased at all sites (lumbar spine, femur, radius). In all clinical subgroups, the mean Z-scores were significantly decreased at radius. The mean Z-scores in premenopausal women were significantly decreased comparatively to postmenopausal women at lumbar spine and femur. In a group of PHPT females matched to controls for age and BMI, only BMD values at radius were lower in PHPT patients than in control (P < 0.03). However, from the whole body scan data, all sites but no trunk were significantly involved in PHPT patients (P < 0.04). Using QUS measurements at calcaneus, the BUA but not SOS in PHPT females was significantly lower (P = 0.03) than in controls. Our results suggest that low BMD at lumbar spine and femur is encountered preferentially in premenopausal women. The BMD decrease predominates at limbs in PHPT with presumably a gradient from proximal to distal part of the limbs. Indeed, the distal part of the limbs are the most affected areas in PHPT whatever the amount of cortical or trabecular bone. PMID- 15893951 TI - Analysis of fibrous proteins from electron microscopy images. AB - This paper considers an approach for analyzing fibrillar collagen structures from electron microscopy images. It enables the quantitative comparison between collagen structural data (electron-optical data) and chemical data. The particular objectives of the paper are to model the electron microscopy images according to the periodic structure of collagen, provide methods for extracting periodic features directly from the experimental data and propose schemes for comparing these features with the theoretical amino-acid distributions of the examined collagen tissue. Theoretical models in the form of sequence-generated histograms are used as reference for extracting and analyzing the structural unit in images from collagen fibrils. In this respect, collagen provides a valuable model system for studying the chemical basis of ultra-structure and the mechanisms of various treatments on a protein, as well as detecting the alterations in collagen fibril structure produced by a disorder. The algorithms developed in this study can be applied to any fibrous protein, provided that its amino acid sequences and structural properties are known. Several application examples are presented. The algorithmic results are compared with clinical studies as to verify the applicability and potential of the proposed methodology. PMID- 15893952 TI - Ca(2+)-dependent proteolysis in muscle wasting. AB - Skeletal muscle wasting is a prominent feature of cachexia, a complex systemic syndrome that frequently complicates chronic diseases such as inflammatory and autoimmune disorders, cancer and AIDS. Muscle wasting may also develop as a manifestation of primary or neurogenic muscular disorders. It is now generally accepted that muscle depletion mainly arises from increased protein catabolism. The ubiquitin-proteasome system is believed to be the major proteolytic machinery in charge of such protein breakdown, yet there is evidence suggesting that Ca(2+) dependent system, lysosomes and, in some conditions at least, even caspases are involved as well. The role of Ca(2+)-dependent proteolysis in skeletal muscle wasting is reviewed in the present paper. This system relies on the activity of calpains, a family of Ca(2+)-dependent cysteine proteases, whose regulation is complex and not completely elucidated. Modulations of Ca(2+)-dependent proteolysis have been associated with muscle protein depletion in various pathological contexts and particularly with muscle dystrophies. Calpains can only perform a limited proteolysis of their substrates, however they may play a critical role in initiating the breakdown of myofibrillar protein, by releasing molecules that become suitable for further degradation by proteasomes. Some evidence would also support a role for lysosomes and caspases in muscle wasting. Thus it cannot be excluded that different intracellular proteolytic systems may coordinately concur in shifting muscle protein turnover towards excess catabolism. Many different signals have been proposed as potentially involved in triggering the enhanced protein breakdown that underlies muscle wasting. How they are transduced to initiate the hypercatabolic response and to activate the proteolytic pathways remains largely unknown, however. PMID- 15893953 TI - Cerebrovascular segmentation from TOF using stochastic models. AB - In this paper, we present an automatic statistical approach for extracting 3D blood vessels from time-of-flight (TOF) magnetic resonance angiography (MRA) data. The voxels of the dataset are classified as either blood vessels or background noise. The observed volume data is modeled by two stochastic processes. The low level process characterizes the intensity distribution of the data, while the high level process characterizes their statistical dependence among neighboring voxels. The low level process of the background signal is modeled by a finite mixture of one Rayleigh and two normal distributions, while the blood vessels are modeled by one normal distribution. The parameters of the low level process are estimated using the expectation maximization (EM) algorithm. Since the convergence of the EM is sensitive to the initial estimate of the model parameters, an automatic method for parameter initialization, based on histogram analysis, is provided. To improve the quality of segmentation achieved by the proposed low level model especially in the regions of significantly vascular signal loss, the high level process is modeled as a Markov random field (MRF). Since MRF is sensitive to edges and the intracranial vessels represent roughly 5% of the intracranial volume, 2D MRF will destroy most of the small and medium sized vessels. Therefore, to reduce this limitation, we employed 3D MRF, whose parameters are estimated using the maximum pseudo likelihood estimator (MPLE), which converges to the true likelihood under large lattice. Our proposed model exhibits a good fit to the clinical data and is extensively tested on different synthetic vessel phantoms and several 2D/3D TOF datasets acquired from two different MRI scanners. Experimental results showed that the proposed model provides good quality of segmentation and is capable of delineating vessels down to 3 voxel diameters. PMID- 15893954 TI - trans-Parinaric acid as a versatile spectroscopic label to study ligand binding properties of bovine beta-lactoglobulin. AB - Advantageous spectroscopic properties of the plant derived polyunsaturated trans parinaric acid (tPnA) was demonstrated in obtaining valuable data on the ligand binding characteristics of the lipocalin member bovine beta-lactoglobulin A (BLG A). Titration of the protein with tPnA resulted in the appearance of an intense negative induced circular dichroism (CD) band and bathochromic shift of the ultraviolet (UV) peak of the ligand. The extrinsic optical activity was interpreted by the chiral contribution of the allylic axial CH bonds of tPnA to the pi-pi(*) transition of the planar tetraene chromophore. Analysis of the series of induced CD curves obtained by CD titration experiment indicated the complexation of a single ligand molecule to a uniform protein binding site. Additionally, the dramatic increase of fluorescence intensity of the lactoglobulin bound ligand suggested the hydrophobic nature of the binding site. CD and fluorescence titration data were utilized to calculate the binding constant (K(a)) of which high value ( approximately 10(6)M(-1)) refers to strong protein association of tPnA. pH dependent reversible dis- and reappearance of the induced CD signal unambigously proved the inclusion of tPnA into the central hydrophobic cavity of the lactoglobulin governed by the protonation induced conformational movement of the EF loop at the opening of the calyx. This conclusion was supported and complemented by molecular docking calculations. PMID- 15893955 TI - Photophysical studies of some dyes in aqueous solution of triton X-100. AB - The spectral (both absorption and fluorescence) and photoelectrochemical studies of a few selective dyes, namely, anionic erythrosin B, neutral riboflavin and cationic safranin O have been carried out in aqueous solution of triton X-100, a neutral surfactant. The results show that the ionic dyes, erythrosin B and safranin O form 1:1 electron donor-acceptor (EDA) or charge-transfer (CT) complexes with triton X-100 both in the ground and excited states, whereas neutral dye riboflavin in its excited state forms 1:1 complex with triton X-100. In these complexes, the dyes act as electron acceptors whereas triton X-100 acts as an electron donor. The fluorescence spectra of erythrosin B and safranin O in presence of triton X-100 show enhancement of fluorescence intensity with red and blue shifts respectively while riboflavin shows normal quenching of fluorescence. A good correlation has been found among photovoltage generation of the systems consisting of these dyes and triton X-100, spectral shift due to complex formation and thermodynamic properties of these complexes. PMID- 15893956 TI - Expanding the frontiers of existing antiviral drugs: possible effects of HIV-1 protease inhibitors against SARS and avian influenza. AB - When unexpected diseases such as the severe acute respiratory syndrome (SARS) and avian influenza become a serious threat to public health, an immediate response is imperative. This should take into consideration existing licensed antiviral drugs against other viral diseases already known to be safe for use in humans. In this report, evidence is presented that HIV-1 protease inhibitors (PIs) currently used in anti-HIV-1 therapies might exert some effects on SARS and perhaps, on avian influenza. Evidence for the potential benefits of PIs against the SARS coronavirus (SARS-CoV) is provided by empirical clinical studies, in vivo viral inhibition assays and computational simulations of the docking of these compounds to the active site of the main SARS-CoV protease. As suggested by in silico docking of these molecules to a theoretical model of a subunit of type A influenza virus RNA-dependent RNA polymerase, there also exists a remote possibility that these PIs may have an effect on avian influenza viruses. Although this evidence is still far from being definitive, the results so far obtained suggest that PIs should be seriously taken into consideration for further testing as potential therapeutic agents for SARS and avian influenza. PMID- 15893957 TI - A real-time Taqman method for hepatitis C virus genotyping. AB - BACKGROUND: There is the need for a rapid, inexpensive method for genotyping hepatitis C virus (HCV) to support clinical practice. OBJECTIVES: To develop a real-time (Rotor-Gene 3000) Taqman assay for HCV genotyping in a single tube. STUDY DESIGN: Seven type-specific probes, two for genotypes 1-3 and one for genotype 4 were designed around genotype-specific motifs in the 5' non-coding (NC) region to create two panels of probes. The first panel included two probes for genotype 1 detection and a single probe each for genotypes 2 and 3. The second panel had two probes for confirmation of genotypes 2 and 3 and a first line probe for genotype 4 detection. A comparative analysis of the Taqman assay against our in-house sequence-based method using 154 consecutive clinical samples, from HCV carriers in Cambridge, and four samples from the Quality Control for Molecular Diagnostics (QCMD) System was undertaken. RESULTS: 158 samples were analysed by conventional sequencing: 49% (n=78) were genotype 1, 11% (n=18) genotype 2, 30% (n=47) genotype 3 and 6% (n=10) genotype 4. For two samples, the sequence data was heterogeneous and difficult to analyse, suggesting mixed infection and for three samples, the viral load was insufficient for sequencing. Concordant results were obtained with the novel Taqman assay for 77/78 (99%) of genotype 1 isolates (positive with both genotype 1 probes), 17/18 (94%) of genotype 2 isolates, 43/47 (91%) of genotype 3 isolates and 10/10 (100%) genotype 4 isolates. One isolate, untypeable with sequencing was genotyped with the Taqman assay. CONCLUSIONS: The Taqman assay was sensitive, specific and reliable over a wide range of viral loads and could identify mixed infections. These results highlight the potential of the Taqman assay as a fast, accurate and convenient method for routine HCV genotyping. PMID- 15893958 TI - Down-regulated expression of PPARalpha target genes, reduced fatty acid oxidation and altered fatty acid composition in the liver of mice transgenic for hTNFalpha. AB - The present study investigated the hepatic regulation of fatty acid metabolism in hTNFalpha transgenic mice. Reduced hepatic mRNA levels and activities of carnitine palmitoyltransferase-II (CPT-II) and mitochondrial HMG-CoA synthase were observed, accompanied by decreased fatty acid oxidation, fatty acyl-CoA oxidase and fatty acid synthase (FAS) activities and down-regulated gene expression of mitochondrial acetyl-CoA carboxylase 2 (ACC2). The mRNA levels of peroxisome proliferator-activated receptor alpha (PPARalpha) and PPARdelta were reduced. The hepatic fatty acid composition was altered, with increased amounts of saturated and polyunsaturated fatty acids. The relative amounts of Delta(9) desaturated fatty acids were decreased, as was Delta(9)desaturase mRNA. The CPT-I mRNA level remained unchanged. The PPARalpha targeted genes CPT-II and HMG-CoA synthase are potential regulators of mitochondrial fatty acid oxidation and ketogenesis in hTNFalpha transgenic mice, and the increased propionyl-CoA level found is a possible inhibitor of these processes. Reduced mitochondrial and peroxisomal fatty acid oxidation may explain the increased hepatic triglyceride level induced by TNFalpha. This is not due to de novo fatty acid synthesis as both FAS activity and gene expression of ACC2 were reduced. PMID- 15893959 TI - Negotiating compliance in heart failure: remaining issues and questions. AB - Living with heart failure (HF) means living with a chronic illness characterized by periods of acute decompensation alternating with periods of relative stability. Improved medical care for patients with cardiovascular diseases, coupled with the aging of the populations in the developed world, has resulted in a steadily increasing prevalence of HF. Rehospitalization rates are high for this patient population. In 20-64% of the cases, poor compliance by patients with the prescribed HF treatment is a contributing factor to hospitalization. This article uses a review of the literature on HF non-compliance, including the prevalence, barriers, consequences, and the long-term outcomes of non-compliance with HF therapy, to illustrate remaining issues and questions. Original studies published in English or German between 1966 and June 2004 identified by combining patient compliance, non-compliance, adherence, self-care, rehospitalization, patient education, and management programs, with heart failure in the search strategy are included. Creative approaches to achieving a true partnership between providers and patients are needed if clinical outcomes are to improve. PMID- 15893960 TI - Treatment of experimental immune complex glomerulonephritis by sodium alginate. AB - We have studied the therapeutic efficacy of the sodium alginate in experimental immune complex glomerulonephritis. Bovine serum albumin (BSA) nephritis was induced in rats by a subcutaneous immunization and daily intravenous administration of BSA. Sodium alginate at two different doses (25 and 50 mg/kg) was administered intraperitoneally at regular 72-h intervals for 6 weeks. Onset of treatment was day 42. Urinary protein was measured weekly and serum anti-BSA antibody was assessed by ELISA method at different intervals. Animals were euthanized at the 12th experimental week and blood samples and kidney specimens were obtained. BUN, serum creatinine and serum cholesterol and triglyceride were measured at the time of sacrifice. Kidney specimens were processed for light and immunofluorescent microscopic examination. The tolerability and inhibitory effect of LVA on matrix metalloproteinase 2 (MMP-2) were tested using WEHI-164 cell line and zymography method. Results of this experiment showed that treatment with sodium alginate could significantly reduce the urinary protein excretion and serum creatinine in treated rats vs. nontreated controls. Anti-BSA antibody titers were lower in treated rats than in controls at the 12th week post immunization. There was no significant difference in the level of BUN and serum lipids between two groups. Whereas, glomerular hypercellularity, PMN infiltration and glomerular deposition of BSA were less intense in treated rats vs. controls. Moreover, in vitro examinations revealed that treatment with LVA, as a very safe agent could diminish MMP-2 activity. These results suggest that treatment with sodium alginate as a new immunosuppressive agent can reduce proteinuria, inhibit MMP-2 activity and suppress the antibody production as well as the development of glomerular lesions in a rat model of immune complex glomerulonephritis. PMID- 15893961 TI - Separation and detection of vitellogenin in fish plasma by capillary zone electrophoresis. AB - A method for coupling capillary zone electrophoresis (CZE) with rapid membrane chromatography purification (RMCP) was established for the analysis of vitellogenin (VTG) in male fish plasma induced with 17ss-estrodiol. CZE analyses of purified VTG were performed in a buffer containing 25 mM sodium borate (pH 8.4). A 50 microm i.d. fused-silica capillary was used for separation and the detection was carried out by UV-diode array at 214 nm. Inter- and intra-assay variabilities of the proposed method were less than 10.06 and 1.95%, respectively. The method has good linear relationship over the scope of 15-2250 microg/ml with a correlation coefficient of R2 = 0.9965 and a detection limit of 7.0 microg/ml. The established CZE method was also applied to directly separate and identify VTG from fish plasma. The results indicated this method could minimize interferences from plasma proteins, allowing the detection of at least 62.5 microg/ml of VTG proteins in total proteins. This is a rapid and easy method to determine the quantity and purity of VTG compared to Bradford method and SDS PAGE. PMID- 15893962 TI - A simple liquid chromatographic method based on intramolecular excimer-forming derivatization and fluorescence detection for the determination of tyrosine and tyramine in urine. AB - A liquid chromatographic (LC) method for sensitive and selective fluorometric determination of p-hydroxyphenylethylamino group containing compounds is described. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butanoyl chloride, followed by reversed-phase LC. The analytes, containing an amino moiety and a phenolic hydroxyl moiety in a molecule, were converted to the corresponding dipyrene labeled derivatives by one-step derivatization. The dipyrene-labeled derivatives afforded intramolecular excimer fluorescence (440-540 nm), which can clearly be discriminated from the normal fluorescence (360-420 nm) emitted from reagent blanks. The derivatives of tyrosine and tyramine could be separated by reversed phase LC on ODS column under conditions of isocratic elution. The detection limits (signal-to-noise ratio = 3) for tyrosine and tyramine were 4.5 and 2.6 fmol per 20 microL injection, which corresponded to analyte concentrations of 0.9 and 0.5 nM, respectively. PMID- 15893963 TI - Liquid chromatographic tandem mass spectrometric determination of five coccidiostats in poultry eggs and feed. AB - A method is described which permits the quantitative detection of the chemical coccidiostats halofuginone, robenidine, diclazuril, nicarbazin and dimetridazole and its main metabolite 2-hydroxydimetridazole in poultry eggs and feed. Sample preparations were kept very simple and are based upon extraction with an organic solvent. Sample extracts were injected into the liquid chromatography tandem mass spectrometry (LC-MS/MS) system on a C18 column and a gradient elution was performed. Dimetridazole-D3 and diclazuril-bis, a structural analogue of diclazuril, were used as internal standards. Detection was performed on a triple quadrupole mass spectrometer in the selected reaction monitoring mode after ionisation in the positive or negative electrospray ionisation mode. Argon was applied as collision gas for collision induced dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC [Official Journal of the European Communities L221 (2002) 8]. PMID- 15893964 TI - Determination of thymidine phosphorylase activity by a non-radiochemical assay using reversed-phase high-performance liquid chromatography. AB - Thymidine phosphorylase (TP) catalyses the conversion of thymidine into thymine. A non-radiochemical assay procedure for TP was developed in which thymine was detected at 265 nm after separation with reversed-phase HPLC. A complete separation of thymidine and thymine was achieved in 6 min and the minimum amount of thymine that could be detected was 0.8 pmol. The assay was linear with reaction times, up to at least 4 h, and protein concentrations up to at least 65 microg/ml. Population analysis showed no differences in TP activity between man and women or with increasing age. PMID- 15893965 TI - Involvement of placental peptidases associated with renin-angiotensin systems in preeclampsia. AB - Preeclampsia is characterized by pregnancy-induced hypertension accompanied with protein urea and generalized edema. Preeclampsia develops during the second half of pregnancy and resolves postpartum promptly, implicating the placenta as a primary cause in the disorder. Normal pregnancy is associated with reductions in arterial pressure and attenuated pressor response to exogenous infused angiotensin II (ANG II). In contrast, women with preeclampsia show the similar sensitivity to the pressor effect of ANG II as do non-pregnant women. To elucidate the involvement of placental peptidases associated with renin angiotensin systems, we determined the localization of angiotensin-converting enzyme (ACE) and aminopeptidase A (AP-A), ANG II degrading enzyme, in the placenta and compared the expression of mRNA and protein in uncomplicated and preeclamptic placenta. In addition, AP-A expression in trophoblastic cells treated with ANG II and ACE expression in HUVECs under hypoxic condition were analyzed, respectively. The expression of both peptidases in the preeclamptic placenta was significantly higher than those from uncomplicated. ACE was primarily localized to venous endothelial cells of stem villous whereas AP-A expression was recognized in the trophoblast and pericytes of fetal arterioles and venules within stem villous. Hypoxia induced ACE expression in HUVECs while both hypoxia and ANG II evoked AP-A expression in trophoblast. These results suggested that hypoxic condition in preeclampsia induces ACE activation in feto placental unit to maintain the fetal hemodynamics and placental AP-A plays a role as a component of the barrier of ANG II between mother and fetus. PMID- 15893966 TI - Stability of proteins: temperature, pressure and the role of the solvent. AB - We focus on the various aspects of the physics related to the stability of proteins. We review the pure thermodynamic aspects of the response of a protein to pressure and temperature variations and discuss the respective stability phase diagram. We relate the experimentally observed shape of this diagram to the low degree of correlation between the fluctuations of enthalpy and volume changes associated with the folding-denaturing transition and draw attention to the fact that one order parameter is not enough to characterize the transition. We discuss in detail microscopic aspects of the various contributions to the free energy gap of proteins and put emphasis on how a cosolvent may either enlarge or diminish this gap. We review briefly the various experimental approaches to measure changes in protein stability induced by cosolvents, denaturants, but also by pressure and temperature. Finally, we discuss in detail our own molecular dynamics simulations on cytochrome c and show what happens under high pressure, how glycerol influences structure and volume fluctuations, and how all this compares with experiments. PMID- 15893967 TI - Prediction of response to infliximab in Crohn's disease. PMID- 15893968 TI - Antisecretory drugs, Helicobacter pylori infection and symptom relief in GORD: still an unexplored triangle. PMID- 15893969 TI - The NLRR gene family and mouse development: Modified differential display PCR identifies NLRR-1 as a gene expressed in early somitic myoblasts. AB - During vertebrate embryogenesis, the somites form by segmentation of the trunk mesoderm, lateral to the neural tube, in an anterior to posterior direction. Analysis of differential gene expression during somitogenesis has been problematic due to the limited amount of tissue available from early mouse embryos. To circumvent these problems, we developed a modified differential display PCR technique that is highly sensitive and yields products that can be used directly as in situ hybridisation probes. Using this technique, we isolated NLRR-1 as a gene expressed in the myotome of developing somites but not in the presomitic mesoderm. Detailed expression analysis showed that this gene was expressed in the skeletal muscle precursors of the myotome, branchial arches and limbs as well as in the developing nervous system. Somitic expression occurs in the earliest myoblasts that originate from the dorsal lip in a pattern reminiscent of the muscle determination gene Myf5, but not at the ventral lip, indicating that NLRR-1 is expressed in a subset of myotome cells. The NLRR genes comprise a three-gene family encoding glycosylated transmembrane proteins with external leucine-rich repeats, a fibronectin domain, an immunoglobulin domain and short intracellular tails capable of mediating protein-protein interaction. Analysis of NLRR-3 expression revealed regulated expression in the neural system in developing ganglia and motor neurons. NLRR-2 expression appears to be predominately confined to the adult. The regulated embryonic expression and cellular location of these proteins suggest important roles during mouse development in the control of cell adhesion, movement or signalling. PMID- 15893970 TI - Activation of Akt (PKB) and suppression of FKHRL1 in mouse and rat oocytes by stem cell factor during follicular activation and development. AB - Although communications between mammalian oocytes and their surrounding granulosa cells mediated by the Kit-Kit ligand (KL, or stem cell factor, SCF) system have been proven to be crucial for follicular development, Kit downstream signaling pathways in mammalian oocytes are largely unknown. In this study, by using ovaries and isolated oocytes from postnatal mice and rats, we demonstrated for the first time that components of the PI3 kinase pathway, the serine/threonine kinase Akt (PKB) which enhances cellular proliferation and survival, and an Akt substrate FKHRL1 which is a transcription factor that leads to apoptosis and cell cycle arrest, are expressed in mammalian oocytes. By using an in vitro oocytes culture system, we found that oocytes-derived Akt and FKHRL1 are regulated by SCF. Treatment of cultured oocytes with SCF cannot only rapidly phosphorylate and activate Akt, but also simultaneously phosphorylate and may therefore functionally suppress FKHRL1, through the action of PI3 kinase. Together with our in situ hybridization and immunohistochemistry data that Akt and FKHRL1 are mostly expressed in oocytes in primordial and primary ovaries and reports that FKHRL1 gene-deficient mice exhibited excessive activation from primordial to primary follicles as well as enlarged oocyte sizes, we suppose that in mammalian oocytes, actions of granulosa cell derived SCF on primordial to primary follicle transition and subsequent follicle development may involve activation of Akt and inhibition of FKHRL1 activities in oocytes. The role of oocyte's Akt may be to enhance follicle development and the role of oocyte's FKHRL1 may be to inhibit follicle development. We propose that the cascade from granulosa cell SCF to oocyte Kit-PI3 kinase-Akt-FKHRL1 may play an important role to regulate the growth rate of mammalian oocytes and hypothetically also the oocyte secretion of factors that may regulate the activation and early development of ovarian follicles. PMID- 15893971 TI - Embryonic deregulation of muscle stress signaling pathways leads to altered postnatal stem cell behavior and a failure in postnatal muscle growth. AB - PW1 is a mediator of p53 and TNFalpha signaling pathways previously identified in a screen to isolate muscle stem cell regulators. We generated transgenic mice carrying a C-terminal deleted form of PW1 (DeltaPW1) which blocks p53-mediated cell death and TNFalpha-mediated NFkappaB activation fused to the myogenin promoter. Embryonic/fetal muscle development appears normal during transgene expression, however, postnatal transgenic pups display severe phenotypes including runtism, reduced muscle mass and fiber diameters resembling atrophy. Atrogin-1, a marker of skeletal muscle atrophy, is expressed postnatally in transgenic mice. Electron microscopic analyses of transgenic muscle reveal a marked decrease in quiescent muscle satellite cells suggesting a deregulation of postnatal stem cells. Furthermore, transgenic primary myoblasts show a resistance to the effects of TNFalpha upon differentiation. Taken together, our data support a role for PW1 and related stress pathways in mediating skeletal muscle stem cell behavior which in turn is critical for postnatal muscle growth and homeostasis. In addition, these data reveal that postnatal stem cell behavior is likely specified during early muscle development. PMID- 15893972 TI - Oocyte nucleus controls progression through meiotic maturation. AB - We analyzed progression through the meiotic maturation in oocytes manipulated to replace the prophase oocyte nucleus with the nucleus from a cumulus cell, a pachytene spermatocyte or the pronucleus from a fertilized egg. Removal of the oocyte nucleus led to a significant reduction in histone H1 kinase activity. Replacement of the oocyte nucleus by a pronucleus followed by culture resulted in premature pseudomeiotic division and occasional abnormal cytokinesis; however, histone H1 kinase activity was rescued, microtubules formed a bipolar spindle, and chromosomes were condensed. In addition to the anomalies observed after pronuclear transfer, those after transfer of the nucleus from a cumulus cell or spermatocyte included a dramatically impaired ability to form the bipolar spindle or to condense chromosomes, and histone H1 kinase activity was not rescued. Expression of a cyclin B-YFP in enucleated oocytes receiving the cumulus cell nucleus rescued histone H1 kinase activity, but spindle formation and chromosome condensation remained impaired, indicating a pleiotropic effect of oocyte nucleus removal. However, when the cumulus cell nucleus was first transformed into pronuclei (transfer into a metaphase II oocyte followed by activation), such pronuclei supported maturation after transfer into the oocyte in a manner similar to that of normal pronuclei. These results show that the oocyte nucleus contains specific components required for the control of progression through the meiotic maturation and that some of these components are also present in pronuclei. PMID- 15893973 TI - Dynamic rearrangement of telomeres during spermatogenesis in mice. AB - Chromosomal structure within the nucleus influences various biological processes such as transcription and replication. Telomeres are located at the end of eukaryotic chromosomes and they can be a decisive factor for correct chromosomal positioning. To gain new insight into telomere dynamics, we examined telomere length and positional changes during spermatogenesis using improved fluorescence in situ hybridization (FISH) and in situ telomeric repeat amplification protocols (TRAP) on histological sections. FISH revealed telomere length and chromosome position within nuclei change dynamically. Telomere extension occurred during spermiogenesis. In situ TRAP analysis verified elevated telomerase activity in elongating spermatids. Together, these data show that elongated spermatids have longer telomeres than precursor spermatogenic cells. This observation indicates that telomere elongation in haploid cells occurs after meiosis and in the absence of genomic replication. Analyses of testes from telomerase null mice further support the significance of telomere dynamics during spermatogenesis and the existence of an alternative telomere extension pathway. PMID- 15893974 TI - Phylogenetic footprinting and genome scanning identify vertebrate BMP response elements and new target genes. AB - The complex gene regulatory networks governed by growth factor signaling are still poorly understood. In order to accelerate the rate of progress in uncovering these networks, we explored the usefulness of interspecies sequence comparison (phylogenetic footprinting) to identify conserved growth factor response elements. The promoter regions of two direct target genes of Bone Morphogenetic Protein (BMP) signaling in Xenopus, Xvent2 and XId3, were compared with the corresponding human and/or mouse counterparts to identify conserved sequences. A comparison between the Xenopus and human Vent2 promoter sequences revealed a highly conserved 21 bp sequence that overlaps the previously reported Xvent2 BMP response element (BRE). Reporter gene assays using Xenopus animal pole ectodermal explants (animal caps) revealed that this conserved 21 bp BRE is both necessary and sufficient for BMP responsiveness. We combine the same phylogenetic footprinting approach with luciferase assays to identify a highly conserved 49 bp BMP responsive region in the Xenopus Id3 promoter. GFP reporters containing multimers of either the Xvent2 or XId3 BREs appear to recapitulate endogenous BMP signaling activity in transgenic Xenopus embryos. Comparison of the Xvent2 and the XId3 BRE revealed core sequence features that are both necessary and sufficient for BMP responsiveness: a Smad binding element (SBE) and a GC-rich element resembling an OAZ binding site. Based on these findings, we have implemented genome scanning to identify over 100 additional putative target genes containing 2 or more BRE-like sequences which are conserved between human and mouse. RT-PCR and in situ analyses revealed that this in silico approach can effectively be used to identify potential BMP target genes. PMID- 15893975 TI - Developmental genetic basis for the evolution of pelvic fin loss in the pufferfish Takifugu rubripes. AB - Paired appendages were a key developmental innovation among vertebrates and they eventually evolved into limbs. Ancient developmental control systems for paired fins and limbs are broadly conserved among gnathostome vertebrates. Some lineages including whales, some salamanders, snakes, and many ray-fin fish, independently lost the pectoral, pelvic, or both appendages over evolutionary time. When different taxa independently evolve similar developmental morphologies, do they use the same molecular genetic mechanisms? To determine the developmental genetic basis for the evolution of pelvis loss in the pufferfish Takifugu rubripes (fugu), we isolated fugu orthologs of genes thought to be essential for limb development in tetrapods, including limb positioning (Hoxc6, Hoxd9), limb bud initiation (Pitx1, Tbx4, Tbx5), and limb bud outgrowth (Shh, Fgf10), and studied their expression patterns during fugu development. Results showed that bud outgrowth and initiation fail to occur in fugu, and that pelvis loss is associated with altered expression of Hoxd9a, which we show to be a marker for pelvic fin position in three-spine stickleback Gasterosteus aculeatus. These results rule out changes in appendage outgrowth and initiation genes as the earliest developmental defect in pufferfish pelvic fin loss and suggest that altered Hoxd9a expression in the lateral mesoderm may account for pelvis loss in fugu. This mechanism appears to be different from the mechanism for pelvic loss in stickleback, showing that different taxa can evolve similar phenotypes by different mechanisms. PMID- 15893976 TI - A POU factor binding site upstream of the Chx10 homeobox gene is required for Chx10 expression in subsets of retinal progenitor cells and bipolar cells. AB - Retinal progenitor cells (RPCs) undergo a series of changes over time that affect their competency to produce different cell types at different times in development. The transcriptional machinery that regulates these changes, as well as associated gene expression changes, have not been characterized. An analysis of the regulatory region of the retinal homeodomain transcription factor, Chx10, was carried out using in ovo electroporations in chick and transgenic mice. An RPC enhancer was defined that mediates reporter activity in subsets of RPCs and directs high-level expression in intermediate and late RPCs. Using bioinformatic and biochemical analysis, a key binding site in this enhancer was found and was shown to be bound by the POU domain factors, Brn-2 and Tst-1/SCIP, in retinal extracts. Analysis of the Brn-2 expression pattern shows that it is expressed in intermediate and late RPCs, but not early RPCs, and thus partially overlaps with expression of the reporter activated by the defined Chx10 enhancer. Biochemical analysis also revealed binding of both Chx10 and Brn-2 to an enhancer of the CNS progenitor cell marker, Nestin. Nestin expression in the retina is restricted to intermediate/late RPC subsets, and genetic evidence is presented that demonstrates that Chx10 represses Nestin expression in early RPCs. A bipolar cell enhancer for Chx10 also was defined, and a role for Brn-2 in expression of Chx10 in bipolar cells is predicted. These data identify Brn-2 as a new marker of subsets of RPCs and suggest a mechanism by which a combination of POU factors and Chx10 define RPC gene expression patterns, such as that of Nestin. PMID- 15893977 TI - Regulation of the lmo2 promoter during hematopoietic and vascular development in zebrafish. AB - The Lmo2 transcription factor, a T-cell oncoprotein, is required for both hematopoiesis and angiogenesis. To investigate the fate of lmo2-expressing cells and the transcriptional regulation of lmo2 in vivo, we generated stable transgenic zebrafish that express green fluorescent protein (EGFP) or DsRed under the control of an lmo2 promoter. A 2.5-kb fragment contains the cis-regulatory elements required to recapitulate endogenous lmo2 expression in embryonic hematopoietic and vascular tissues. We further characterized embryonic Lmo2+ cells through transplantation into vlad tepes (vlt), an erythropoietic mutant. These Lmo2+ primitive wave donor cells differentiated into circulating hematopoietic cells and extended the life span of vlt recipients, but did not demonstrate long-term repopulation of the erythroid lineage. Promoter analysis identified a 174-bp proximal promoter that was sufficient to recapitulate lmo2 expression. This element contains critical ETS-binding sites conserved between zebrafish and pufferfish. Furthermore, we show that ets1 is coexpressed with lmo2, and overexpression experiments indicate that ets1 can activate the lmo2 promoter through this element. Our studies elucidate the transcriptional regulation of this key transcription factor, and provide a transgenic system for the functional analysis of blood and blood vessels in zebrafish. PMID- 15893978 TI - Allocation and specification of the genital disc precursor cells in Drosophila. AB - The adult structures of Drosophila melanogaster are derived from larval imaginal discs, which originate as clusters of cells within the embryonic ectoderm. The genital imaginal disc is composed of three primordia (female genital, male genital, and anal primordia) that originate from the embryonic tail segments A8, A9, and A10, respectively, and produce the sexually dimorphic genitalia and analia. We show that the genital disc precursor cells (GDPCs) are first detectable during mid-embryogenesis as a 22-cell cluster in the ventral epidermis. Analysis of mutant and double mutant phenotypes of embryonic patterning genes in the GDPCs, together with their expression patterns in these cells, revealed the following with respect to the origins and specification of the GDPCs. The allocation of the GDPCs from the ventral epidermis requires the function of ventral patterning genes, including the EGF receptor and the spitz group of genes. The ventral localization of the GDPCs is further restricted by the action of dorsal patterning genes. Along the anterior-posterior axis, several segment polarity genes (wingless, engrailed, hedgehog, and patched) are required for the proper allocation of the GDPCs. These segment polarity genes are expressed in some, but not all of the GDPCs, indicating that anterior and posterior compartments are not fully established in the GDPCs. In addition, we found that the three primordia of the larval genital disc have already been specified in the GDPCs by the coordinated actions of the homeotic (Hox) genes, abdominal-A, Abdominal-B, and caudal. By identifying how these different patterning networks regulate the allocation and primordial organization of the 22 embryonic precursors of the compound genital disc, we demonstrate that at least some of the organization of the larval disc originates as positional information in the embryo, thus providing a context for further studies on the development of the genital disc. PMID- 15893979 TI - Brn1/2/4, the predicted midgut regulator of the endo16 gene of the sea urchin embryo. AB - A specific prediction of our detailed cis-regulatory analysis of the Strongylocentrotus purpuratus (Sp) endo16 gene was that the later expression of this gene would be driven by a midgut-specific transcriptional regulator. We have now identified this factor and determined some of its functions. The cDNA sequence reveals it to be a POU domain factor related closely to the mammalian factors Brain-1, -2, and -4. The factor was termed SpBrn1/2/4 (henceforth Brn1/2/4). Quantitative measurements of transcript prevalence show that the gene is first activated in the 20-h blastula, but there remain only about 100 molecules of brn1/2/4 mRNA per embryo (only a few per endoderm cell) until an abrupt 10-fold increase occurs as gastrulation begins. Measured in the same embryos, the late rise in prevalence of endo16 transcripts follows that of brn1/2/4 transcripts. As predicted by the endo16 model, brn1/2/4 expression is confined perfectly to the midgut, coincident with the domain of endo16 expression. The kinetics of accumulation of these transcripts indicates that the switch into the late phase of endo16 expression occurs when the brn1/2/4 transcript level nears its plateau (2000 molecules mRNA per embryo), after which each endo16 gene produces about 1 mRNA every 2 min (about 380 molecules mRNA per min in the whole embryo). Arrest of Brn1/2/4 translation by MASO treatment blocks the late phase of endo16 expression and specifically abolishes expression of cis regulatory Module B of endo16, while not affecting Module A, also as predicted. The brn1/2/4 gene lies downstream of the regulatory genes executing post gastrular specification of the midgut, as shown by further gene expression perturbation experiments which provide an initial glimpse of the underlying network architecture. PMID- 15893980 TI - An ancient transcriptional regulatory linkage. AB - Changes in gene regulatory networks are a major engine for creating developmental novelty during evolution. Conversely, regulatory linkages that survive for very long evolutionary periods might be characteristic of ancient and abstract functions of fundamental utility to all metazoans. The proneural genes, which encode a distinctive family of basic helix-loop-helix (bHLH) transcriptional activators, act to promote neural cell fates in the ectoderm of diverse species. Here we report that these genes have been associated for at least 600-700 million years--since before the cnidarian/bilaterian divergence--with a high-affinity binding site for Hairy/Enhancer of split (Hes) repressor proteins. We suggest that the systematic identification of such ancient and conserved connections will be a powerful means of uncovering the primordial functions of transcription factors and signaling systems. PMID- 15893981 TI - Impact of transcription factor Sox8 on oligodendrocyte specification in the mouse embryonic spinal cord. AB - The myelin-forming oligodendrocytes of the mouse embryonic spinal cord express the three group E Sox proteins Sox8, Sox9, and Sox10. They require Sox9 for their specification from neuroepithelial cells of the ventricular zone and Sox10 for their terminal differentiation and myelination. Here, we show that during oligodendrocyte development, Sox8 is expressed after Sox9, but before Sox10. Loss of Sox8 did not impair oligodendrocyte specification by itself, but enhanced the Sox9-dependent defect. Oligodendrocyte progenitors were still generated in the Sox9-deficient spinal cord, albeit at 20-fold lower rates than in the wildtype. Combined loss of Sox8 and Sox9, in contrast, led to a near complete loss of oligodendrocytes. Other cell types such as ventricular zone cells and radial glia remained unaffected in their numbers as well as their rates of proliferation and apoptosis. Oligodendrocyte development thus relies on the differential contribution of all three group E Sox proteins at various phases. PMID- 15893982 TI - A novel hes5/hes6 circuitry of negative regulation controls Notch activity during neurogenesis. AB - HES transcriptional repressors are important components of the Notch pathway that regulates neurogenesis from Drosophila to vertebrates. These proteins are normally induced by Notch activity and inhibit neural commitment by antagonizing the activity of proneural genes. We describe here four chick hes genes that are expressed during neurogenesis: three hes5-like genes (hes5-1, hes5-2 and hes5-3) and one hes6-like (hes6-2). We show that hes6-2 represses transcription of the hes5 genes, thus functioning as a negative regulator of Notch signaling. Conversely, hes6-2 may be repressed by hes5 activity. In cells committing to differentiation, we find that hes6-2 is up-regulated by proneural genes and contributes to the proneural program of neuronal commitment by preventing Notch activity in these cells. In neural progenitors, Notch signaling produces an initial burst of hes5 activity, which represses hes6-2. However, as hes5 transcription declines due to negative auto-regulation, hes6-2 may become active and inhibit the remaining hes5 activity to end Notch signaling. These cells can then enter a new cycle of fate decisions and will be kept as progenitors if a new pulse of Notch activity occurs. Maintenance of progenitors during vertebrate neurogenesis therefore requires that these cells go through successive cycles of Notch activity. We propose that the hes5/hes6 circuitry of negative cross regulations is a conserved feature of the Notch pathway that underlies these cycles in neural progenitors. PMID- 15893983 TI - Evolution of T lymphocytes and cytokine expression in classical swine fever (CSF) virus infection. AB - This study characterized the cell-mediated immune response in pigs inoculated with the Alfort 187 isolate of classical swine fever (CSF) virus. Quantitative changes in the T-lymphocyte population (CD3(+), CD4(+) and CD8(+)) and qualitative changes in cytokine expression (IL-2, IL-4 and IFNgamma) by these cells in serum, thymus and spleen were demonstrated. These changes coincided spatially and temporally with previously described quantitative and qualitative changes in monocyte-macrophage populations, thus demonstrating the contribution of the two cell populations to lymphoid depletion. Moreover, examination of cytokine expression in thymus and spleen samples revealed a type 1 cell-mediated immune response in the early and middle stages of the experiment, giving way to a type 2 immune response towards the end of the experiment; these findings, which accorded with the serological results and lymphopenia, may influence the delayed humoral response characteristic of CSF. PMID- 15893984 TI - Distribution of leucocyte subsets in the canine respiratory tract. AB - Histochemistry and immunohistochemistry were used to characterize leucocyte subsets in the respiratory tract of 15 outbred dogs (five aged <6 months and 10 aged >1 year) that had no evidence of respiratory disease. No organized nose- or bronchus-associated lymphoid tissue was observed in any of the sections examined. IgA(+) plasma cells predominated in nasal mucosa and in all parts of the bronchial tree, with fewer IgG(+) and IgM(+) plasma cells. The numbers of IgA(+) and IgM(+) cells were significantly greater in the nasal mucosa than in any other part of the respiratory mucosa. There were significantly fewer IgA(+), IgG(+) and IgM(+) cells in all parts of the respiratory tract in the puppies than in the adults. The number and distribution of mast cells and cells expressing MHC class II, L1 or CD1c were recorded. Mast cells were mainly found in the subepithelial lamina propria of nasal and bronchial mucosa and in the alveolar interstitium, and cells expressing IgE had a similar distribution. Mast cells were also present within muscle layers of the bronchial tree. The numbers of mast cells and MHC class II(+) cells were significantly greater in the nasal mucosa than in any other part of the respiratory mucosa. In the nose, carina and primary and secondary bronchus, there were significantly more mast cells and MHC class II(+) cells in puppies than in adult dogs, whereas the numbers of L1(+) cells and CD1c(+) cells in most sites were significantly greater in older dogs. There were significantly more CD3(+) and CD8(+) cells in the nasal mucosa than in any part of the bronchial mucosa. In most parts of the respiratory mucosa, CD4(+), CD8(+) and TCR alphabeta(+) cells were present in significantly greater numbers in adults than in puppies. All parts of the respiratory tract had similar numbers of mucosal CD4(+) and CD8(+) T lymphocytes. TCR gammadelta(+) cells were absent or sparse in all samples. These data, obtained from dogs without respiratory disease, will enable comparisons to be made with dogs suffering from infectious or inflammatory nasal, bronchial and pulmonary diseases. PMID- 15893985 TI - Expression of prion protein in the gut of mice infected orally with the 301V murine strain of the bovine spongiform encephalopathy agent. AB - Transmissible spongiform encephalopathies (TSEs) are characterized by the accumulation of an abnormal, disease-associated prion protein (PrP(d)). Expression of its normal cellular counterpart (PrP(c)) by the host is a pre requisite for the spread of infection to the central nervous system and the development of disease. Moreover, cells expressing PrP(c) at specific sites such as the gastrointestinal tract might be regarded as the initial point of PrP(c) PrP(d) conversion after infection by the oral route. In this study, inbred mice of the I/M strain were infected orally with the 301V murine strain of the bovine spongiform encephalopathy agent. The expression of PrP(c) and the accumulation of PrP(d) in the intestine was then investigated immunohistochemically, together with the variations in immunoreactivity that resulted from different pretreatments of the tissue. After proteinase K (PK) pretreatment, abnormal PrP was still detectable only in the gut-associated lymphoid tissue (GALT) of clinically affected mice and, to a much more limited degree, in the enteric nervous system (ENS). Cellular PrP that disappeared after PK treatment was particularly conspicuous in the ENS and present to a lesser extent in the GALT of all mice examined after inoculation with 301V or with normal brain homogenates, as well as in uninoculated controls. These findings suggested that not all PrP found in infected mice was PrP(d) and that part of the PrP(d) was sensitive to PK treatment. Reactivity to PrP antibody 1A8 was consistently found in the absorptive epithelium of the intestinal villi, with or without PK pretreatment. However, epithelial immunolabelling was comparable in inoculated and uninoculated mice and was also consistently seen in PrP "knockout" mice used as controls. It is therefore concluded that immunohistochemically detectable accumulation of PrP(d) in the gut of mice is a relatively late event in the pathogenesis of experimental infection in this model and that the immunoreactivity observed in the intestinal epithelium does not correspond to PrP expression. While enterocytes may still play a role in the uptake of infection from the intestinal lumen, the results do not suggest that these cells are a site of initial accumulation of PrP(d). PMID- 15893986 TI - An immunohistochemical study of canine nasal aspergillosis. AB - In this study, histochemistry and immunohistochemistry were used to characterize the phenotype and distribution of leucocytes in the distal nasal mucosa of 15 dogs with nasal aspergillosis. The most consistent histopathological finding was a severe, predominantly lymphoplasmacytic, inflammatory infiltration of the lamina propria. Fungal hyphae were not observed to invade the mucosa but were found at the mucosal surface and within material collected from the nasal cavity. The main immunohistochemical findings were (1) a predominance of IgG(+) plasma cells over IgA(+) and IgM(+) plasma cells, (2) significant numbers of macrophages and dendritic cells expressing MHC class II molecules, (3) macrophages and neutrophils expressing L1 antigen and (4) a mixture of CD4(+) and CD8(+) T cells. These findings are consistent with a dominant Th1-regulated cell-mediated immune response. The nature of the inflammatory infiltrate and the lack of invasiveness of the mucosa by the fungus, together with the clinical course of the disease and the apparent immunocompetence of the affected dogs, suggest that canine nasal aspergillosis resembles the chronic erosive non-invasive fungal sinusitis described in human patients. PMID- 15893987 TI - Proinflammatory cytokines induce lymphocyte apoptosis in acute African swine fever infection. AB - Twenty-one pigs inoculated with a highly virulent isolate (E70) of African swine fever (ASF) virus were killed 1-7 days later; a further three animals served as uninfected controls. An early increase in TNF-alpha, IL-1alpha, IL-1beta and IL-6 expression was detected in lymphoid organs from infected animals, together with an increase in the serum concentrations of TNF-alpha and IL-1beta. These changes were accompanied by increased apoptosis of lymphocytes, and the presence of infected and uninfected macrophages showing changes indicative of secretory and phagocytic activation. The present study demonstrated an increase in the number of macrophages expressing TNF-alpha, IL-1 and IL-6 in proximity to lymphocytes undergoing apoptosis, supporting previous suggestions that in acute ASF proinflammatory cytokines induce lymphocyte apoptosis. PMID- 15893988 TI - Renal lipofuscinosis in Danish slaughter cattle. AB - A study was performed to characterize dark brown or black discoloured kidneys ("black kidneys") in Danish slaughter cattle and to investigate the aetiology and pathogenesis. In 133 939 cattle entering four abattoirs, 359 cases of "black kidneys" were recorded. Of these, 57 cases were submitted for macroscopical, microscopical, and ultrastructural examination. A pigment with characteristics similar to those of lipofuscin was found in secondary lysosomes in epithelial cells of the proximal tubules. Pigment accumulation was the cause of discoloration, with a positive correlation between the discoloration of the renal cortex and the degree of pigment accumulation. Cases occurred only in cattle of the Holstein breed or the Red Danish Dairy breed and mainly in animals aged 3 years or older. In these breeds, prevalences of 0.44% and 2.51% were found, respectively. Epidemiological analyses indicated that affected animals aged 4.5 to 6.5 years or 7.5 to 8.5 years were culled more frequently than unaffected cattle. Epidemiological and genealogical analyses strongly indicated a genetic aetiology with simple autosomal recessive inheritance. PMID- 15893989 TI - Filter function and immune complex trapping in splenic ellipsoids. AB - The role of splenic ellipsoids in the trapping of particulate material and immune complexes was investigated in mink (Mustela vison). The ellipsoids were prominent, with typical features such as a permeable endothelium and a discontinuous basement membrane surrounded by a sheath of macrophages and reticular cells. Ellipsoidal trapping of circulating particles was demonstrated 10 min after intracardiac injection of colloidal carbon and fluorescent microspheres. Preformed peroxidase-antiperoxidase immune complexes were detected in ellipsoids 10 min and also 1 h after intracardiac injection. Erythrocytes were frequently observed in the ellipsoidal sheath, and many phagocytized fragments of erythrocytes were found in the ellipsoidal macrophages. It was concluded that mink ellipsoids are effective blood filters with a role in retention of circulating particulate material, and that mammalian splenic ellipsoids also have the ability to trap immune complexes. PMID- 15893990 TI - Antigenicity of ovine strains of Anaplasma phagocytophilum grown in tick cells and ovine granulocytes. AB - Antigens prepared from ovine granulocytes and tick cells infected with ovine strains of Anaplasma phagocytophilum, the causative agent of tick-borne fever, were tested in respect of their suitability for the assay of antibodies in ovine sera by enzyme-linked immunosorbent assay (ELISA) and by indirect immunofluorescent antibody test (IFAT). Antigens prepared from tick cells were as sensitive and specific as those expressed in ovine granulocytes for the detection of specific antibodies by ELISA, but they failed to react in the IFAT with immune sera obtained from sheep previously infected with ovine strains of A. phagocytophilum. PMID- 15893991 TI - Cyclin D3 expression in normal fetal, normal adult and neoplastic feline tissue. AB - Cyclin D3 is a tightly regulated cell cycle protein and member of the cyclin D family-a group of proteins that facilitates the progression of a cell through G(1) and into the S phase of the cell cycle. All cells use at least one of the cyclin D proteins for cell cycle regulation. In this study, feline tissues (normal fetal and adult, and neoplastic) were examined immunohistochemically for expression and topographical distribution of cyclin D3. Its distribution was similar to that in human tissues in health and neoplasia, and suggested a dual role of cyclin D3 in cell proliferation and differentiation. Immature lymphoid tissue and proliferating epithelial cells in health and neoplasia were immunoreactive for cyclin D3, whereas expression of the protein in other immunoreactive tissues reflected differentiated cell types. Immunoreactivity for cyclin D3 was particularly striking in germinal centre cells of normal lymph nodes and B-cell lymphomas, and in normal suprabasal epithelial cells of the skin and mucous membranes of the oropharynx and in squamous cell carcinomas at these sites. PMID- 15893992 TI - Primary endocardial malignant spindle-cell sarcoma in the right atrium of a dog resembling a malignant peripheral nerve sheath tumour. AB - An unusual malignant spindle-cell sarcoma in the right atrium of a 7-year-old male hunting terrier is described. The neoplasm arose from the endocardium, protruded into the lumen of the right atrium, was covered with endothelium, and showed local invasive growth into the atrial wall. The tumour was composed of interlacing bundles of spindle cells, sometimes arranged in whorls resembling the Antoni type A pattern. The extracellular matrix showed abundant reticulin fibres. Immunohistochemistry revealed an intense labelling of tumour cells for vimentin, and a partial labelling for neuron-specific enolase, S100 protein, and Melan-A. The morphology of the tumour indicated a primary malignant peripheral nerve sheath tumour, resembling a neoplasm described in rats. PMID- 15893993 TI - Ectodysplasin-1 deficiency in a German Holstein bull associated with loss of respiratory mucous glands and chronic rhinotracheitis. AB - A 2-year-old German Holstein bull was identified as a carrier of a mutation within the X-chromosomal ED1 gene, which encodes a TNF-related signalling molecule mainly involved in ectodermal development. The clinicopathological appearance was associated with hypotrichosis, hypodontia, and a reduced number of eccrine glands, in addition to chronic rhinotracheitis and partial squamous metaplasia. Furthermore, for the first time in an ED1-deficient animal, a complete lack of respiratory mucous glands was observed. This suggests that the ED1 gene plays a role in the development of mucous glands, the absence of which resembles a feature of X-linked anhidrotic ectodermal dysplasia (ED1) in human patients. PMID- 15893994 TI - Clinicopathological features of globoid cell leucodystrophy in cats. AB - Clinical and pathological findings consistent with globoid cell leucodystrophy (GLD) were evaluated in two domestic shorthaired cats, aged 3 and 4 months. Both showed neurological signs mainly characterized by progressive pelvic limb ataxia, paraplegia with loss of deep pain perception in the pelvic limb, and intentional tremors of the thoracic limbs. Pathological changes affecting the central and peripheral nervous systems were characterised by diffuse, bilateral and symmetrical myelin loss, and marked astrogliosis. In the leucodystrophic areas there was perivascular accumulation of large PAS-positive, non-metachromatic macrophages (globoid cells), with intracytoplasmic accumulation of crystalloid tubular aggregates. Peripheral nerves showed demyelinating features with thin myelin sheaths, myelin splitting, and ballooning; the nerve fibres had bizarre shapes due to the presence of pale inclusions in the Schwann cells. GLD in cats shares clinical and pathological features with the disease described in other animals and human beings. The neurological signs differed from those of other feline inborn neurometabolic diseases and cerebellar hypoplasia. PMID- 15893995 TI - Characterization of stretch-activated calcium permeable cation channels in freshly isolated myocytes of the cricket (Gryllus bimaculatus) lateral oviduct. AB - Stretch-activated channels (SACs) were investigated in myocytes isolated from the lateral oviduct in cricket Gryllus bimaculatus using the cell-attached or excised inside-out patch clamp technique. Application of both negative and positive pressure (10-100 cm H(2)O) into the patch pipettes induced the unitary channel current openings. The open probability (NPo) of the channel increased when negative pressure applied into the patch pipettes increased. The single channel conductance for this channel was approximately 20 pS with 140 mM Na(+), K(+), or Cs(+) in the patch pipettes and was approximately 13 pS with 100mM Ca(2+) or Ba(2+) in the patch pipettes. External application of Gd(3+), La(3+), Cd(2+) and Zn(2+)inhibited the channel with the IC(50) values of 14, 15, 28, and 18 microM respectively. Interestingly external application of TEA, a specific blocker of K(+) channel, also inhibited this channel with IC(50) value of 8.8mM. These results show for the first time the presence of stretch activated Ca(2+) permeable nonselective cation channel in myocytes isolated from the cricket lateral oviduct. The physiological significance of this channel in oviposition behavior is discussed. PMID- 15893996 TI - Heartbeat patterns during the postembryonic development of Drosophila melanogaster. AB - Pulsations of the dorsal vessel were recorded in vivo during the whole postembryonic development of D. melanogaster, by means of a newly invented, pulse light opto-cardiographic method. The young larvae of the 1st and 2nd instars submerged in the feeding medium exhibited extremely high rates of heartbeat, 7Hz at room temperature. These values are among the highest rates of heartbeat ever recorded in the animal kingdom. The fully grown larvae of the 3rd instar showed approximately half of the maximum heartbeat rate (3.5-4Hz), which became stabilized after pupariation to 2.5-2.7Hz. The larval heartbeat was always uni directional, in the forward-oriented or anterograde direction and it was almost continuous. The slowly disintegrating, old larval heart used to beat at the constant frequency of 2.5-2.7Hz until complete cessation of all cardiac functions in 1-day-old puparium. In spite of the persisting constant heartbeat frequency, the transformation process of the larval heart was associated with successively decreasing amplitude of the systolic contractions and with the prolongation of the resting periods. The newly formed heart of the pupal-adult structure exhibited a qualitatively new pattern of heartbeat activity, which was manifested by periodic reversal of the heartbeat with the faster anterograde and slower retrograde phases. The frequencies of both of these reciprocal cardiac pulsations gradually increased during the advanced pharate adult period, reaching the values of 4-5Hz at the time of adult eclosion. Adult males and females also exhibited a perfect pattern of heartbeat reversal, with still very high rates of the anterograde heartbeat, in the range of 5-6Hz. In addition to the cardiac functions, we have recorded several kinds of extracardiac pulsations, which often interfered severely with the recordings of the heartbeat. There were strong, irregular extracardiac pulsations of a neurogenic nature (somatic muscles, oral armature) and relatively slow extracardiac pulsations of a myogenic nature (intestinal peristaltics, 0.2-0.3Hz). The extracardiac and cardiac pulsations were independent, their functions were not correlated. A possibility of creating new challenges in combination of molecular biology with the functional physiology of the heart have been discussed. PMID- 15893997 TI - Parasitization of Manduca sexta larvae by the parasitoid wasp Cotesia congregata induces an impaired host immune response. AB - During oviposition, the parasitoid wasp Cotesia congregata injects polydnavirus, venom, and parasitoid eggs into larvae of its lepidopteran host, the tobacco hornworm, Manduca sexta. Polydnaviruses (PDVs) suppress the immune system of the host and allow the juvenile parasitoids to develop without being encapsulated by host hemocytes mobilized by the immune system. Previous work identified a gene in the Cotesia rubecula PDV (CrV1) that is responsible for depolymerization of actin in hemocytes of the host Pieris rapae during a narrow temporal window from 4 to 8h post-parasitization. Its expression appears temporally correlated with hemocyte dysfunction. After this time, the hemocytes recover, and encapsulation is then inhibited by other mechanism(s). In contrast, in parasitized tobacco hornworm larvae this type of inactivation in hemocytes of parasitized M. sexta larvae leads to irreversible cellular disruption. We have characterized the temporal pattern of expression of the CrV1-homolog from the C. congregata PDV in host fat body and hemocytes using Northern blots, and localized the protein in host hemocytes with polyclonal antibodies to CrV1 protein produced in P. rapae in response to expression of the CrV1 protein. Host hemocytes stained with FITC labeled phalloidin, which binds to filamentous actin, were used to observe hemocyte disruption in parasitized and virus-injected hosts and a comparison was made to hemocytes of nonparasitized control larvae. At 24h post-parasitization host hemocytes were significantly altered compared to those of nonparasitized larvae. Hemocytes from newly parasitized hosts displayed blebbing, inhibition of spreading and adhesion, and overall cell disruption. A CrV1-homolog gene product was localized in host hemocytes using polyclonal CrV1 antibodies, suggesting that CrV1-like gene products of C. congregata's bracovirus are responsible for the impaired immune response of the host. PMID- 15893998 TI - Characterization of two midgut proteinases of Helicoverpa armigera and their interaction with proteinase inhibitors. AB - Two serine proteinases from the midgut of Helicoverpa armigera have been partially purified and characterized. One proteinase, HGP-1, was capable of hydrolyzing a synthetic substrate of elastase and was inhibited by elastatinal. The second proteinase, HGP-2, was inhibited by a trypsin inhibitor. Molecular weights of HGP-1 and HGP-2 were approximately 26.0 and 29.0kDa, respectively. Both the proteinases exhibited alkaline pH optima in the range of 10-11. Furthermore, interaction of HGP-1 and HGP-2 with proteinase inhibitors (PIs) from host and non-host plants was studied. HGP-1 was not only insensitive to a PI from chickpea (host) but was also able to degrade it. The same PI from chickpea was able to inhibit over 50% activity of HGP-2. On the contrary, PIs from potato (non host) showed strong inhibition of both, HGP-1 and HGP-2 and also demonstrated protection of chickpea seed proteins from digestion by both the HGPs. These results could provide important clues in designing strategies for sustainable use of plant PIs in developing insect-tolerant transgenic plants. PMID- 15893999 TI - Regulation of JH epoxide hydrolase versus JH esterase activity in the cabbage looper, Trichoplusia ni, by juvenile hormone and xenobiotics. AB - JH III esterase and JH III epoxide hydrolase (EH) in vitro activity was compared in whole body Trichoplusia ni homogenates at each stage of development (egg, larva, pupa and adult). While activity of both enzymes was detected at all ages tested, JH esterase was significantly higher than EH activity except for day three of the fifth (last) stadium (L5D3). For both enzymes, activity was highest in eggs. Adult virgin females had 4.6- and 4.0-fold higher JH esterase and EH activities, respectively, than adult virgin males. JH III metabolic activity also was measured in whole body homogenates of fifth stadium T. ni that were fed a nutritive diet (control) or starved on a non-nutritive diet of alphacel, agar and water. With larvae that were starved for 6, 28 and 52 h, EH activity per insect equivalent was 48%, 5% and 1%, respectively, of the control insects. At the same time points, JH esterase activity levels in starved T. ni were 29%, 4% and 3% of that of insects fed the nutritive diet. Selected insect hormones and xenobiotics were administered topically or orally to fifth stadium larvae for up to 52 h, and the effects on whole body EH and JH esterase activity analyzed. JH III increased the JH III esterase activity as high as 2.2-fold, but not the JH III EH activity. The JH analog, methoprene, increased both JH esterase and EH activity as high as 2.5-fold. The JH esterase inhibitor, 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP), had no impact on EH activity. The epoxides trans- and cis-stilbene oxide (TSO and CSO) in separate experiments increased the EH activity approximately 2.0 fold. TSO did not alter JH esterase levels when topically applied, but oral administration reduced activity to 70% of the control at 28 h, and then increased the activity 1.8-fold at 52 h after the beginning of treatment. CSO had no effect on JH esterase activity. Phenobarbital increased EH activity by 1.9-fold, but did not change JH esterase levels. Clofibrate and cholesterol 5alpha,6alpha-epoxide had no effect on EH. JH esterase activity also was not affected by clofibrate, but cholesterol 5alpha,6alpha-epoxide reduced the JH esterase activity to 60-80% of the control. The biological significance of these results is discussed. PMID- 15894000 TI - Impact of male mating history on the temporal sperm dynamics of Choristoneura rosaceana and C. fumiferana females. AB - In the oblique-banded leafroller, Choristoneura rosaceana, and the spruce budworm, C. fumiferana, male reproductive performance decreases with consecutive matings. While the onset time of mating did not vary, the time spent mating was longer in mated than in virgin males. Furthermore, a decline observed in the spermatophore mass with successive matings was associated with a concomitant decline in its apyrene and eupyrene spermatozoa content. In the hours following mating, spermatozoa migrate from the spermatophore, located in the bursa copulatrix, to the spermatheca. Regardless of the male's previous mating history, the number of apyrene sperm dropped rapidly in the days following mating whereas the number of eupyrene spermatozoa declined gradually. As the temporal pattern of sperm movement was similar in all treatments, females mated with previously-mated males would suffer from sperm shortage sooner than those mated with virgins. Large C. rosaceana females stored more apyrene spermatozoa in their spermatheca than small ones, irrespective of the time after mating or male mating history, while only large females mated with once-mated males received more apyrene sperm and accessory gland secretions than small ones mated with virgin or twice-mated males. The results obtained in this study are discussed in relation with their potential impact on the reproductive success of both sexes. PMID- 15894001 TI - Effects of adult-derived carbohydrates, amino acids and micronutrients on female reproduction in a fruit-feeding butterfly. AB - It is generally believed that butterflies (and other holometabolous insects) rely primarily on reserves accumulated during the larval stage for reproduction, whereas the carbohydrate-rich adult diet is thought to mainly cover energy requirements. In at least some species though, realization of the full reproductive potential is extensively affected by post-eclosion nutrition. While the importance of carbohydrates is fairly well understood, the role of adult derived amino acids and micronutrients is controversial and largely unknown, respectively. We here focus on the effects of different adult diets on female reproduction in the tropical, fruit-feeding butterfly Bicyclus anynana (Nymphalidae). Carbohydrates were the most important adult-derived nutrients affecting reproduction. Adding amino acids, vitamins or minerals to sucrose-based solutions did not yield a reproductive output equivalent to that of fruit-fed females, which showed the highest performance throughout. This suggests that either not yet identified compounds of fruit substantially contribute to reproduction, or that resource congruence (the use of nutrient types in a specified ratio) rather than any specific nutrient component is of key importance. Apart from adult income, realized fecundity depended on egg size and longevity, with the former dominating when dietary quality was low, but the latter when quality was high. Thus, the egg size-number trade-off seems to be affected by female nutrition. PMID- 15894002 TI - Clustering of adhesion receptors following exposure of insect blood cells to foreign surfaces. AB - Cell-mediated immune responses of insects involve interactions of two main classes of blood cells (hemocytes) known as granular cells and plasmatocytes. In response to a foreign surface, these hemocytes suddenly transform from circulating, non-adherent cells to cells that interact and adhere to each other and the foreign surface. This report presents evidence that during this adhesive transformation the extracellular matrix (ECM) proteins lacunin and a ligand for peanut agglutinin (PNA) lectin are released by granular cells and bind to surfaces of both granular cells and plasmatocytes. ECM protein co-localizes on cell surfaces with the adhesive receptors integrin and neuroglian, a member of the immunoglobulin superfamily. The ECM protein(s) secreted by granular cells are hypothesized to interact with adhesion receptors such as neuroglian and integrin by cross linking and clustering them on hemocyte surfaces. This clustering of receptors is known to enhance the adhesiveness (avidity) of interacting mammalian immune cells. The formation of ring-shaped clusters of these adhesion receptors on surfaces of insect immune cells represents an evolutionary antecedent of the mammalian immunological synapse. PMID- 15894003 TI - Prolonged maintenance of water balance by adult females of the American spider beetle, Mezium affine Boieldieu, in the absence of food and water resources. AB - Moisture requirements were evaluated for female adults of spider beetles Mezium affine Boieldieu and Gibbium aequinoctiale Boieldieu to determine how they are differentially adapted for life in a dry environment. Features showing extreme desiccation resistance of M. affine were an impermeable cuticle wherein activation energies (43kJ/mol) were suppressed, daily water losses as little as 0.3%/day with an associated group effect, a low 64% water content and an impressive ability to survive nearly 3 months with no food and water. Behaviorally, the extended period of water stress and fasting was marked by long intervals of physical inactivity (quiescence), as though dead. These characteristics emphasizing water retention rather than gain are shared by G. aequinoctiale and reflect a typical xerophilic water balance profile. Water uptake was restricted to imbibing liquid, as evidenced by uptake of dye-stained droplets of free water and a critical equilibrium activity of 1.00a(v), where the inability to absorb water vapor from the air fails to equilibrate declining water levels (gain not equal to loss) except at saturation. Four-fold reduction in survival time within dry air and accelerated water loss rates with high activation energies for female adults of the closely related winged Prostephanus truncatus (Say) suggest that the enhanced water conservation of spider beetles is due, in part, to fusion of their elytra supplemented by entering into quiescence. PMID- 15894004 TI - Cyclic AMP affects the haemocyte responses of larval Galleria mellonella to selected antigens. AB - Signal transduction of the innate immediate responses of insect haemocytes to foreign matter is rarely considered. Herein using a combination of adenylate cyclase inhibitors and activators and phosphodiesterase inhibitors we determined that cyclic adenosine monophosphate (cAMP) at high levels normally impairs non self response. Haemocyte contact with glass and bacteria lowered cAMP in vitro. Inactive phosphodiesterases, including type 4, impaired haemocyte reactions in vitro. Using the drugs in vivo to modulate adenylate cyclase and phosphodiesterases altered the total and types of haemocytes. Adenylate cyclase inhibitors and etazolate (a type 4 phosphodiesterase inhibitor) alone produced changes in the haemograms similar to those caused by Bacillus subtilis. Sequential injections of an enzyme modulator followed by B. subtilis impaired bacterial removal due (1) in the case of enzyme inhibitors, to the removal of haemocytes prior to bacterial challenge and (2) in the case of forskolin and IBMX to the shut-down of the haemocytes. Activating adenylate cyclase or inhibiting phosphodiesterase impaired bacterial removal when co-injecting the compounds and bacteria. PMID- 15894005 TI - Plasmatocyte sensitivity to plasmatocyte spreading peptide (PSP) fluctuates with the larval molting cycle. AB - Plasmatocyte spreading peptide (PSP) is a cytokine from the moth Pseudoplusia includens that activates a class of hemocytes called plasmatocytes to bind and spread on foreign surfaces. Previous structure-function studies on PSP used plasmatocytes collected from P. includens larvae that were in the late stages of the last (fifth) instar. Here, we report that plasmatocyte sensitivity to PSP varied significantly during the fourth and fifth instar. PSP weakly activated plasmatocytes early in the instar when hemolymph juvenile hormone (JH) titers were relatively high and ecdysteroid titers were low, but strongly activated plasmatocytes late in the instar after JH titers declined and ecdysteroid titers rose. In contrast, plasmatocytes did not vary in their response to plasma, which contains other factors besides PSP that affect plasmatocyte function. In vitro assays indicated that 20-hydroxyecdysone (20E) dose-dependently synergized PSP activity, whereas the JH analog methoprene antagonized PSP activity. Methoprene had no effect on adhesion and spreading of granular cells, but plasmatocytes from larvae topically treated with methoprene exhibited a reduction in sensitivity to PSP. Collectively, these results indicate that plasmatocyte sensitivity to PSP fluctuates in relation to the molting cycle, and that PSP activity is affected by juvenoids and ecdysone. PMID- 15894006 TI - Lanthanide-based luminescent assays for ligand-receptor interactions. AB - The evaluation of receptor ligand interactions is important in the field of drug discovery and development. Currently these interactions are typically measured with cumbersome (low throughput) radiolabels. Higher throughput screens are available such as fluorescent measurements of G-protein coupled receptor-induced Ca2+ increases or fluorescence anisotropy, yet these have limited applicability and/or low signal to noise. Hence, there is a need to develop more widely applicable and more sensitive labels that can be used to monitor ligand-receptor interactions. Lanthanides provide an attractive alternative to the traditional labels used for monitoring ligand-receptor interactions. The incorporation of lanthanide labels into traditional assays used to assess receptor-ligand interactions can make these assays more affordable, less time consuming and amenable to automation. Lanthanides can be coupled to ligands and provide strong luminescent signals that can be detected using time-resolved fluorescence (TRF) methods. This approach takes advantage of the long fluorescence lifetime of the lanthanide and can detect less than one attomole of europium in a multiwell plate sample. This short review provides a basic introduction into lanthanides and TRF and describes some of the recent assays which have utilized lanthanides as labels to assess ligand-receptor interactions. PMID- 15894007 TI - Effects of phencyclidine on schedule-controlled responding following neurotoxic lesions of the striatum. AB - The effects of phencyclidine on an operant task were evaluated prior to and after neurotoxic lesions of the striatum in rats. Subjects were trained to respond on a fixed-interval 90-second schedule for water presentation. The degree to which phencyclidine disrupted responding was first evaluated (dose range 1.0-4.0 mg/kg). The subjects were then divided into three matched groups and received bilateral intraventricular injections of 6-hydroxydopamine (6-OHDA) (100 microg), kainic acid (0.25 microg), or vehicle delivered stereotaxically. 6-OHDA was used to destroy the presynaptic neurons of the nigro-striatal pathway and kainic acid was employed to destroy the postsynaptic neurons whose cell bodies are located in the striatum. Following recovery, the phencyclidine dose-response curve was repeated in the fixed-interval paradigm. It was observed that 6-OHDA-induced damage resulted in a rightward shift of the dose-response curve indicating tolerance to phencyclidine and caused a significant depletion of striatal dopamine and gamma-aminobutyric acid (GABA). Kainic acid-induced damage resulted in a leftward shift in the dose-response curve indicating sensitivity to the schedule-disruptive effects of phencyclidine and produced a significant GABA depletion. The vehicle-treated rats exhibited no shift in their sensitivity to phencyclidine. These observations indicate that the effects of phencyclidine are mediated, at least in part, by striatal dopaminergic neurons. PMID- 15894008 TI - Electro-acupuncture potentiates the disulphide-reducing activities of thioredoxin system by increasing thioredoxin expression in ischemia-reperfused rat brains. AB - Reactive oxygen species can directly affect the conformation and activity of sulfhydryl-containing proteins by oxidation of their thiol moiety. During the process of ischemia-reperfusion, the thioredoxin (Trx) system (consisting of thioredoxin reductase (TR), Trx and NADPH) prevents susceptible proteins from this oxidative modification. Oxidative damage is one of the most damaging stress in ischemia. If oxidative stress could be minimized, the damage occurred will be minimized accordingly. We therefore investigated whether electroacupuncture (EA) treatment at Fengchi (GB20) or Zusanli (ST36) acupoints in post-ischemic rats could increase TR-related activities and Trx expression which would translate into maintaining the intact thiol moiety of susceptible proteins in the surrounding. Our results indicated that EA treatment at either acupoint increased the Trx expression in ischemic-reperfused brain tissues. Induced Trx expressed levels gradually increased from post-ischemia day 1 to day 4. Statistical analysis revealed that there was no observable difference in the effect of EA treatment at GB20 and ST36. Sham EA treatment did not induce any Trx expression. EA at either acupoint did not alter TR activities in both non-ischemic and ischemic-reperfused rat brains. Taken overall, our finding suggests that EA treatment at GB20 or ST36 could increase Trx expression which could minimize oxidative modifications of thiol groups of surrounding proteins. PMID- 15894009 TI - A novel polysaccharide of black soybean promotes myelopoiesis and reconstitutes bone marrow after 5-flurouracil- and irradiation-induced myelosuppression. AB - The aim of this study was to investigate the promotion of myelopoiesis by an active polysaccharide of black soybean (PSBS). Murine spleen cells were collected from ICR mice and conditioned media (SCM) was prepared by incubating these cells without PSBS (normal-SCM) or with PSBS in concentrations ranging from 12.5 to 100 microg/ml (PSBS-SCM). Murine bone marrow cells were treated with PSBS alone or SCM to induce the formation of colonies, including CFU-GM, CFU-GEMM, BFU-E and HPP-CFC. The concentrations of six hematopoietic growth factors contained in SCM were measured using enzyme-linked immunoassay. In the live animal experiment, PSBS was administered orally to total body-irradiated (TBI) and 5-fluorouracil (5 FU)-treated mice to assess the reconstitution of bone marrow after myelosuppression. PSBS-SCM stimulated CFU-GM, CFU-GEMM, BFU-E and HPP-CFC colony formation with 45.0, 5.0, 6.2 and 6.6-fold increases, respectively. However, neither PSBS alone nor normal-SCM had such a colony-stimulating effect. In PSBS SCM, the levels of IL-6, IL-17, G-CSF and GM-CSF were markedly increased, but not those of IL-3 and SCF. Oral administration of PSBS in mice not only restored the leukocyte counts reduced by TBI and 5-FU treatment but also enhanced CFU-GM colony formation of bone marrow cells without a significant change in body weight. We conclude that PSBS promotes myelopoiesis activity in the bone marrow, stimulates production of various hematopoietic growth factors from spleen cells, and reconstitutes bone marrow that has been myelosuppressed by irradiation and 5 FU. PMID- 15894010 TI - Increased cold allodynia following intrathecal N-methyl-D-aspartate in rats with a mononeuropathy. AB - NMDA receptors are involved in the modulation of neuropathic pain behavior and central sensitization. In vivo, this is mostly demonstrated by the use of specific antagonists such as MK801. Because studies evaluating the direct impact of spinal NMDA in neuropathic pain models are lacking, we performed a series of experiments to study the role of spinal NMDA injection on existing cold allodynia, as a measurement of neuropathic pain behavior in rodents. Intrathecal injection of NMDA resulted in an enhanced neuropathic pain behavior in CCI rats on a cold plate. The activity was present from a dose of 5 ng/rat onward and could selectively be reversed by intraperitoneal injections of doses of > or = 0.01 mg/kg MK801. These results support the regulatory role of NMDA receptors in the hypersensitivity to cold observed in neuropathic pain behavior in rodents. PMID- 15894011 TI - Coronary effects of endothelin-1 and vasopressin during acute hypotension in anesthetized goats. AB - Coronary effects of endothelin-1 and vasopressin during acute hypotension, and the role of NO and prostanoids in these effects were examined in anesthetized goats. Left circumflex coronary artery flow was measured electromagnetically, and hypotension was induced by constriction of the caudal vena cava in animals non treated (7 goats) or treated with the inhibitor of NO synthesis N(w)-nitro-L arginine methyl esther (L-NAME, 5 goats), the cyclooxygenase inhibitor meclofenamate (5 goats) or both drugs (5 goats). Under normotension (22 goats), mean arterial pressure averaged 93 +/- 3 mm Hg and coronary vascular conductance (CVC) 0.37 +/- 0.025 ml/min/mm Hg. Endothelin-1 (0.01-0.3 nmol) and vasopressin (0.03-1 nmol), intracoronarily injected, dose-dependently decreased CVC by up to 56% for endothelin-1 and 40% for vasopressin. During hypotension in every condition tested, mean arterial pressure decreased to approximately 60 mm Hg, and CVC only decreased during hypotension pretreated with L-NAME (23%) or L-NAME + meclofenamate (34%). Under non-treated hypotension, the decreases in CVC by endothelin-1 were augmented approximately 1.5 fold, and those by vasopressin were not modified. This increase in CVR by endothelin-1 was not affected by L-NAME and was reversed by meclofenamate or L-NAME + meclofenamate. The coronary effects of vasopressin were not modified by any of these treatments. Therefore, acute hypotension increases the coronary vasoconstriction in response to endothelin-1 but not to vasopressin. This increased response to endothelin-1 may be related to both inhibition of NO release and release of vasoconstrictor prostanoids. PMID- 15894012 TI - Interrelation between long-chain fatty acid oxidation rate and carnitine palmitoyltransferase 1 activity with different isoforms in rat tissues. AB - This study examined the interrelation between the long-chain fatty acid (LCFA) oxidation rate and the carnitine palmitoyltransferase (CPT) 1 activity in various tissues containing L-CPT1 or M-CPT1. The Liver, kidney, heart, white and red gastrocnemius muscles, and white and brown adipose tissues obtained from Sprague Dawley rats were examined. In the tissues containing L-CPT1 the liver showed a significantly higher (P<0.01) palmitate oxidation rate and CPT1 activity than the kidney. Among the tissues containing M-CPT1, the brown adipose tissue showed the highest palmitate oxidation rate and CPT1 activity. The tissues containing M-CPT1 (r2=0.907, p<0.001) showed a strong positive correlation between the palmitate oxidation rate and the CPT1 activity. The ratios of the palmitate oxidation rate to the CPT1 activity were calculated. The ratio in the liver was highest and the ratio in the kidney was lowest among the tissues. The ratios of the tissues containing M-CPT1 were similar. These results showed that the LCFA oxidation rates in the tissues containing M-CPT1 were directly proportional to the CPT1 activity, but not similarly proportional to the CPT1 activity in the tissues containing L-CPT1. In conclusion, CPT1 activity seems very important factor for LCFA oxidation, but it might be not the only rate-limiting step in LCFA oxidation. PMID- 15894014 TI - Serotonin transporter kinetics in rats selected for extreme values of platelet serotonin level. AB - By selective breeding of Wistar rats for the extreme values of platelet serotonin (5HT) level (PSL), we have developed earlier two sublines of animals differing markedly in this parameter. Further studies, performed on the protein and mRNA levels, revealed platelet serotonin transporter (5HTt) as parameter underlying mentioned differences in PSL between sublines. In this work, we have performed full-kinetic analysis of platelet serotonin uptake (PSU) in animals from the genetically selected sublines. The results demonstrated marked differences in maximal velocity (V(max)) of the 5HT transporter, as contrasted to the lack of any difference in the Michaelis constant (K(m)). High correlation between PSL and V(max) of PSU was demonstrated, revealing that the number of membrane 5HT transporter sites is under genetic control and responsible for marked differences in PSL between high- and low-5HT sublines. These results enabled further selective breeding of animals for the extremes of V(max) of platelet 5HT transporter, and so the development of more specific model "Wistar-Zagreb 5HT rats". PMID- 15894013 TI - Protective effects of glutathione and cysteine on the methylmercury-induced striatal dopamine release in vivo. AB - The possible protective effects of glutathione (GSH), cysteine (CYS) and methionine (MET) on the Methylmercury (MeHg)-induced dopamine (DA) release from rat striatum were investigated using in vivo microdialysis coupled to HPLC with electrochemical detection. Intrastriatal infusion of MeHg 400 microM increased extracellular DA levels to 1941 +/- 199% in terms of basal levels. Infusion of MeHg 400 microM in GSH 400 microM pretreated animals, only increased striatal DA levels to 465 +/- 104%, in terms of basal levels, this increase being 76% lower than induced by MeHg alone. Conversely, the infusion of MeHg 400 microM after infusion of GSH 400 microM increased DA levels to 1019 +/- 96% in terms of basal levels, this increase being 47.5% lower than that observed in MeHg non-pretreated animals. The infusion of MeHg 400 microM in CYS 400 microM -pretreated animals, increased striatal DA levels to 740 +/- 149%, in terms of basal levels, this increase being 62% lower than that induced by MeHg in non-pretreated animals. The infusion of MeHg 400 microM in MET 400 microM pretreated animals increased striatal DA levels to 2011 +/- 230% in terms of basal, an increase that was not significantly different from that produced by MeHg 400 muM alone. In summary, the administration of compounds containing free -SH groups prevented the MeHg-induced DA release from rat striatum, probably due to the binding of MeHg to -SH groups. This would result in a lower metal availability to interact with -SH membrane proteins groups, which would decrease MeHg ability to interact with DA transporter. PMID- 15894017 TI - Canine mycoplasmas. AB - This review aims to summarise our current understanding of the role of mycoplasmas in domestic dogs. Canine mycoplasmology is a small field, with less than 50 publications in the past 40 years. In this time we have gained knowledge about the number of species and have made associations with infections in dogs. However much evidence is still lacking. The importance of all canine mycoplasmas remains unknown, yet certain species are associated with canine anaemia (Mycoplasma haemocanis), respiratory disease (Mycoplasma cynos) and urogenital tract infections (Mycoplasma canis). Mycoplasmas can be isolated in pure culture from canine clinical specimens and it is hoped that this review will stimulate veterinarians to consider mycoplasmas as a potential cause of disease in dogs, especially when antibiotic therapy is failing. PMID- 15894015 TI - Involvement of novel protein kinase C isoforms in carbachol-stimulated insulin secretion from rat pancreatic islets. AB - The roles of protein kinase C (PKC) isoforms in cholinergic potentiation of glucose-induced insulin secretion were investigated in rat pancreatic islets. Western-blot analysis showed the presence of PKC-alpha, betaII, delta, epsilon, eta, and zeta, but not PKC-betaI, gamma, or iota, in the islets. Carbachol (CCh) caused translocations of PKC-alpha, betaII, delta, and epsilon from the cytosol to the plasma membrane. CCh facilitated 7-mM glucose-induced insulin secretion from isolated rat islets. The CCh-stimulated insulin secretion was significantly suppressed by the generic PKC inhibitor chelerythrine. In contrast, Go 6976, an inhibitor of conventional PKC isoforms, had no effect on the insulin secretion stimulated by CCh, although it significantly inhibited that induced by phorbol 12 myristate 13-acetate. These results suggest that the novel PKC isoforms activated by CCh, i.e., PKC-delta and/or epsilon, participate in the stimulatory effect of CCh on insulin secretion. PMID- 15894018 TI - Effect of submaximal exercise on horse homocysteinaemia: possible implications for immune cells. AB - Physical exercise induces a reduction of immune defences and an imbalance of red ox status. In this study plasma levels of cysteine and homocysteine (Hcy) were determined in horses before and after submaximal treadmill exercise as well as the effect on horse lymphocyte proliferation. The exercise induced a significant increase in plasma Hcy levels, which remained high both after the 20 min recovery period and after 2 h of rest. Moreover, a reduction of lymphocyte responsiveness to the proliferative stimulus induced by Concanavalin A was observed. The effects of different Hcy concentrations on the proliferative capacity of lymphocytes in culture were also tested. The results indicated that 10 microM of this amino acid can reduce the proliferative capacity of resting lymphocytes as well as their responsiveness to mitogen. Moreover, our results suggest that homocysteinaemia could be considered one of the parameters affected by physical exercise in horses and that this amino acid could be implicated in the effects of physical exercise on the immune system. PMID- 15894016 TI - Molecular classification of doxazosin-induced alterations in the rat prostate using gene expression profiling. AB - We investigated molecular changes that occurred during chronic administration of doxazosin, an alpha1-adrenoceptor (AR) antagonist, using Affymetrix GeneChip analysis of gene expression. Rats were treated with doxazosin (4 mg/kg/day subcutaneously, supplemented with 4 mg/kg/day orally) for 12 weeks. Labeled cRNA was prepared and the subsequent hybridization to rat 230A arrays was performed. The alterations in gene expression levels of candidate genes identified by microarray analysis with potential biological relevance were verified by real time reverse transcription polymerase chain reaction (RT-PCR) using SYBR Green I. Doxazosin treated rats had significantly heavier prostates compared to control rats. Microarray analysis revealed that chronic doxazosin treatment caused changes in the expression levels of 625 genes, of which 39 were related to cell death, necrosis, growth, proliferation and G-protein signalling pathways in the rat prostate. Furthermore, RT-PCR experiments, in accord with the microarray analysis, indicated that chronic doxazosin treatment caused an up-regulation in the mRNA expression level of clusterin, an antiapoptotic mediator, and epiregulin, a mitogen, in the ventral and dorsolateral prostate, respectively. These findings, that demonstrate chronic doxazosin administration causes significant changes in the expression of several hundred genes in the rat prostate, may provide insight into the long-term efficacy of alpha1-AR antagonists in the treatment of benign prostatic hyperplasia. PMID- 15894019 TI - Serum paraoxonase activity in dairy cows during pregnancy. AB - Preparturient dairy cows are at high risk of metabolic and reproductive disorders and oxidative stress is considered to be involved in these events. We investigated the serum paraoxonase activity in dairy cows during pregnancy and alterations in lipid and lipoprotein patterns in this period. The relation between paraoxonase activity and HDL-cholesterol concentration was also compared. The study was carried out on 76 pregnant lactating and 26 pregnant dry Holstein dairy cows. The serum paraoxonase activity was determined by the method of hydrolysing of paraoxon, while triglyceride, cholesterol and HDL-cholesterol concentrations were measured by the enzymatic kit methods. A significantly higher serum triglyceride concentration (P<0.001) was observed in dry cows compared to lactating cows. The total cholesterol and HDL-cholesterol concentrations were significantly lower (P<0.001) in dry cows than in lactating ones. In dry cows, paraoxonase activity was significantly lower than in those lactating (P<0.001). There was no significant difference in paraoxonase/HDL-cholesterol ratio between the investigated groups. It seems that the lower HDL concentration could be one of the causes of reduced paraoxonase activity considering the role of HDL as a carrier of most paraoxonase molecules in the blood. A decreased serum paraoxonase activity could diminish the effectiveness and total capacity of the whole antioxidative system during prepartum period in dairy cattle. PMID- 15894020 TI - Rat, caprine, equine and bovine erythrocyte ghosts exposed to t-butyl hydroperoxide as a model to study lipid peroxidation using a chemiluminescence assay. AB - The aim of the present study was to analyze the time-course of t-butyl hydroperoxide-induced changes in lipid peroxidation, fatty acid composition and chemiluminescence intensity in rat, caprine, equine and bovine erythrocyte ghosts. A relatively high content of arachidonic acid (C20:4 n6) and docosahexaenoic acid (C22:6 n3) was characteristic of the rat erythrocyte ghosts. The fatty acid composition of native erythrocyte ghosts obtained from caprine, equine and bovine was characterized by a high content of oleic acid (C18:1 n9) and a low content of the peroxidable polyunsaturated fatty acids (C20:4 n6 and C22:6 n3). The proportion of linoleic acid (C18:2 n6) was higher in equine and bovine compared to rat and caprine. Increase in lipid peroxidation in rat erythrocyte ghosts was maximal within 12 min of incubation, t-butyl hydroperoxide concentration dependent and was paralleled by a decrease in C18:2 n6, C20:4 n6 and C22:6 n3 and an increase in chemiluminescence formation. Polyunsaturated fatty acids (PUFAs) present in rat erythrocyte ghosts exhibit the highest sensitivity to oxidative damaged and their sensitivity increases as a power function of the number of double bonds per fatty acid molecule. Light emission in caprine, equine and bovine erythrocyte ghosts was very low, t-butyl hydroperoxide concentration-dependent but changes in fatty acid composition were not observed. The main conclusion of this work is that a low unsaturation degree of fatty acids in erythrocyte ghosts of caprine, equine and bovine prevent the lipid peroxidation on those membranes when they are incubated with t-butyl hydroperoxide. PMID- 15894021 TI - Immunohistochemical studies of scrapie archival material from Irish ARQ/ARQ sheep for evidence of bovine spongiform encephalopathy-derived disease. AB - Since scrapie and bovine spongiform encephalopathy (BSE) in sheep are clinicopathologically indistinguishable, BSE in sheep may have been misdiagnosed as scrapie. Disease-specific prion protein (PrP(d)) patterns in archival tissues of 38 Irish ARQ/ARQ sheep diagnosed as scrapie-affected were compared to those in four Dutch BSE-challenged sheep. When medulla oblongata was immunolabelled with an antibody directed against amino acids 93-99 of ovine prion protein (ovPrP), intraneuronal PrP(d) was apparent in all 38 Irish sheep but was absent in BSE challenged sheep. When lymphoid follicles were immunolabelled with antibodies directed against amino acids 93-106 of ovPrP, granule clusters of PrP(d) were seen in 34 of the 38 Irish sheep. Follicles of the remaining four archive sheep contained either no PrP(d) or single PrP(d) granules, similar to follicles from BSE-challenged sheep. Based on the medulla results, none of the archival cases had BSE-derived disease. The identification of some scrapie sheep with little or no intrafollicular PrP(d) suggests that this technique may be limited in discriminating between the two diseases. PMID- 15894022 TI - Immunotherapeutic potential of Ocimum sanctum (L) in bovine subclinical mastitis. AB - Immunotherapeutic potential of aqueous extract of Ocimum sanctum (O. sanctum) leaf in bovine sub-clinical mastitis (SCM) was investigated. Somatic cell count (SCC), total bacterial count (TBC), milk differential leukocyte count (DLC), phagocytic activity and Phagocytic index and leukocyte lysosomal enzymes like myeloperoxidase and acid phosphatase content were evaluated after intramammary infusion of aqueous leaf extract of O. sanctum. The results revealed that the aqueous extract of O. sanctum treatment reduced the TBC and increased neutrophil and lymphocyte counts with enhanced phagocytic activity and phagocytic index. Similarly, the lysosomal enzymes contents of the milk polymorphonuclear cells (PMNs) were also enhanced significantly in animals treated with the extract. The results suggest that the crude aqueous extract of O. sanctum (leaf) possesses some biologically active principles that are antibacterial and immunomodulatory in nature. As such, the present wok substantiates the therapeutic use of medicinal herb and also emphasizes on the potential of the commonly available non toxic substances to enhance the mammary immunity. PMID- 15894023 TI - Expression and distribution of monocarboxylate transporter 1 (MCT1) in the gastrointestinal tract of calves. AB - In the present study the expression and distribution of monocarboxylate transporter 1 (MCT1) along the gastrointestinal tract (rumen, reticulum, omasum, abomasum, duodenum, jejunum, ileum, cecum and colon) of calves were investigated on both mRNA and protein levels. The expression of MCT1 protein and its distribution were determined by Western blotting and immunohistochemical staining, respectively by using antibody for MCT1. MCT1 protein was visualized as a 43-kDa band on immunoblots of the membrane proteins prepared from the various regions examined, and it was more highly expressed in forestomach and large intestine than in abomasum and small intestine. With the use of reverse transcriptase-polymerase chain reaction, mRNA encoding for MCT1 was demonstrated in the different tissues examined. The immunohistochemical study confirmed the Western blot findings and showed strong MCT1 immunopositive staining in the stratified squamous epithelia of the forestomach as well as the epithelial cells lining the digestive tract in the cecum, proximal colon, and distal colon. The results suggest that MCT1 may play a role in the transport of SCFA and their metabolites in the gastrointestinal tract of bovines. PMID- 15894024 TI - Seroepidemiology of Toxoplasma gondii and Neospora caninum in dogs from the state of Paraiba, Northeast region of Brazil. AB - A cross-sectional study was conducted to determine the seroprevalence of anti Toxoplasma gondii and anti-Neospora caninum antibodies and to investigate the risk factors related to antibodies against T. gondii and N. caninum in dogs of the city of Campina Grande, state of Paraiba, Northeast region of Brazil. For this purpose, 286 blood samples were collected from dogs during the rabies vaccination campaign, in September 2003, and on this occasion questionnaires addressing epidemiological aspects of the infections were given to each dog owner. The sera were analyzed for anti-T. gondii and anti-N. caninum antibodies by indirect fluorescent antibody tests. Of the total of 286 dogs, 129 were positive for T. gondii (titer16) with a prevalence value of 45.1% (95% CI=39.24 51.07%). For N. caninum, 24 dogs were positive (titer50), with a prevalence value of 8.4% (95% CI=5.45-12.23%). Antibodies to T. gondii and N. caninum were found simultaneously in 14 dogs (4.9%; 95% CI=2.7-8.08%). For T. gondii infection, the risk factors associated with seroprevalence was the age of the animals, with dogs older than one year presenting higher values of odds ratio, and co-habitation of cats in the household. For N. caninum infection, dogs that have street contact had higher odds of seropositivity than dogs that remained exclusively in a domestic environment. PMID- 15894025 TI - Do canine parvoviruses affect canine neurons? An immunohistochemical study. AB - In cats (most of which died from panleukopenia), cerebral neurons have recently been shown to be susceptible to canine parvovirus infection. In addition to positive immunostaining and distinct in situ hybridization signals, signs of neurodegeneration were identified by histopathology, mainly in the diencephalic area. Similar histological lesions of the diencephalic regions in dogs have also attracted attention; therefore, an immunohistochemical study was initiated to determine the possible infection of canine neurons with canine parvoviruses. The study was carried out on formalin-fixed and paraffin-embedded brain tissue, with and without signs of neurodegeneration, from 40 dogs, most of them dying from parvovirus enteritis. Immunohistochemistry, using polyclonal antiserum against canine parvoviruses, was negative in all 40 cases, suggesting that, unlike cats, canine parvoviruses do not seem capable of infecting canine neurons. PMID- 15894026 TI - Evaluation of a single dose versus a divided dose regimen of amoxycillin in treatment of Actinobacillus pleuropneumoniae infection in pigs. AB - The theory of a time-dependent effect of amoxycillin was examined in a model of porcine Actinobacillus pleuropneumoniae (Ap)-infection using clinically relevant dosage regimens. Twenty hours after infection of fourteen pigs, when clinical signs of pneumonia were present, one group of pigs received a single dose of amoxycillin (20 mg/kg, i.m.), whereas another group received four doses of 5 mg/kg injected at 8-h intervals. A similar AUC of the plasma amoxycillin concentration versus time curve was obtained in the two groups, whereas the maximum concentration was threefold higher using the single high dose. Plasma amoxycillin was above the MIC for twice as long using the fractionated dosage scheme. The condition of the animals was evaluated by clinical and haematological observations combined with quantification of biochemical infection markers: C reactive protein, zinc and ascorbic acid. Within 48 h of treatment, the pigs in both treatment groups recovered clinically. No significant differences in the time-course of clinical observations or plasma concentrations of the biomarkers of infection were observed between the two treatments. In conclusion, the efficacy of these two dosage regimens of amoxycillin was not significantly different in treatment of acute Ap-infection in pigs. PMID- 15894027 TI - Efficacy of low molecular weight heparin in a canine model of thromboplastin induced acute disseminated intravascular coagulation. AB - The aim of this study was to test the efficacy of different dosages of low molecular weight heparin (LMWH) in acute DIC which was induced in anaesthetised dogs by 4 h infusions of a canine lung thromboplastin extract. In all animals during the first 2 h, development of acute DIC was characterised by decreasing fibrinogen concentrations, platelet numbers, factor V- and antithrombin activities. Two hours after starting the thromboplastin infusion, intravenous LMWH treatment in different dosages started in groups 2 and 3 to achieve plasma levels between 0.27+/-0.01 and 0.36+/-0.02 anti-FXaUml(-1) or 0.62+/-0.08 and 0.90+/-0.07 antiFXaUml(-1) (mean+/-SD), respectively, during the time period of parallel administration of thromboplastin and LMWH (group 1=control; 4 dogs/group). In this time period, changes in factor V activity and fibrinogen concentration did not differ between group 2 and the control group. This was in contrast to group 3. The results of this study indicate that an efficacious interruption of the consumption reaction in cases of severe canine DIC requires high plasma heparin levels. PMID- 15894028 TI - Haematological profile on non-lactating Mediterranean buffaloes (Bubalus bubalis) ranging in age from 24 months to 14 years. AB - Haematological studies were performed on 100 clinically normal non-lactating Mediterranean buffalo species ranging in age from 24 months to 14 years, to determinate the range of normal haematological values for this ruminant species. The animals were divided in 5 groups according to age: Group I, 2-3 years old which had never calved, Group II, 3-4 years old (primipara buffaloes), Group III, 5-7 years old, Group IV 8-10 years old and Group V over 10 years of age. All the haematological values obtained were comparable with the normal values found in adult cattle, and similar to those reported in Indian water buffalo species. The heifer buffalo showed an higher values for packed cell volume (PCV) compared with the older animals, but lower values for mean corpuscular haemoglobin concentration (MCHC) and mean corpuscular haemoglobin (MCH) (P 0.01). In animals above 8 years of age, the white cell count was lower with a significant reduction in absolute values of lymphocytes (P 0.01). Higher absolute values of eosinophils levels was found in the group V (P 0.01). PMID- 15894029 TI - Detection of classical swine fever virus in archival formalin-fixed tissues by reverse transcription-polymerase chain reaction. AB - Classical swine fever (CSF) is an economically important Office International des Epizooties list A disease of swine. Inadequate diagnostic infrastructure in developing countries may sometimes make the conduit of viable samples to diagnostic laboratories difficult and at times formalin-fixed tissues may be the only morbid materials available which are not amenable to most laboratory tests. The potential diagnostic abilities in such situations would be enhanced if a highly sensitive and specific method as reverse transcription-polymerase chain reaction (RT-PCR) could be applied to such samples. In the present study, archival formalin-fixed CSF samples were subjected to successful RT-PCR amplification of a 172-bp fragment from E2/NS2 region of CSFV genome. The fidelity of amplification was established by a dot-blot hybridization assay employing a homologous biotinylated cDNA probe. The optimized procedure may be resorted to as a diagnostic tool if viable samples are unavailable and may have the potential to facilitate retrospective studies on archival samples. PMID- 15894030 TI - Bovine herpesvirus type 1 abortions detected by a semi-nested PCR in Brazilian cattle herds. AB - Bovine herpesvirus type 1 (BHV-1) was investigated by a semi-nested polymerase chain reaction (SN-PCR) and by MDBK cell culture virus isolation in organ fragments from 55 aborted fetuses collected from beef and dairy cattle herds with history of reproductive problems in the North of Parana State, Brazil. A 425 bp amplicon of the BHV-1 glycoprotein D gene was detected in 14 (25.4%) aborted fetuses. BHV-1 was isolated in MDBK cells from the tissue of 5 (9.1%) fetuses. The specificity of positive results was evaluated by Restriction Fragment Length Polymorphism (RFLP) with Bgl I restriction of DNA amplified by SN-PCR, and by virus-neutralization and immunofluorescence with rabbit anti-BHV-1 polyclonal antibodies for virus isolated in cell culture. The results of this work demonstrate the importance of using other diagnostic techniques, like SN-PCR, for BHV-1 detection in organ fragments from aborted fetuses and the high frequency of this virus in reproductive failures in Brazilian cattle herds. PMID- 15894031 TI - Polyhydroxylated steroids from the soft coral Sinularia dissecta. AB - A repeated silica gel column chromatography followed by HPLC purification on the methanol extract of marine soft coral Sinularia dissecta, resulted in the isolation of fifteen polyhydroxylated steroids (1-15), involving six new C-18 functionalized steroids, 3beta-acetoxy-1alpha,11alpha-dihydroxygorgost-5-en-18 oic acid (1), gorgost-5-en-1alpha,3beta,11alpha,18-tetrol (2), 18-acetoxy 1alpha,3beta,11alpha-trihydroxygorgost-5-ene (3), 24(S)-3beta-acetoxy-1alpha, 11alpha-dihydroxyergost-5-en-18-oic acid (4), 24(S)-ergost-5-en 1alpha,3beta,11alpha,18-tetrol (5), and dissectolide (6). The structures of the new compounds were determined on the basis of extensive spectroscopic data (IR, MS, (1)H and (13)C NMR, HMQC, HMBC, and NOESY) analysis. Compound 6 was found as an unusual sterol bearing a lactone functionality. PMID- 15894032 TI - Synthesis of some new steroidal [16alpha,17alpha-d]-isoxazolines. AB - Regioselective synthesis of novel steroidal anti-inflammatory ante drug analogues, viz., [16alpha,17alpha-d]-isoxazolines 1(a-h) and 2(a-h) prepared in a single step in good yield by the reaction of 16-dehydropregnenolone acetate (16 DPA) 1 or related 21-chloro-20-oxopregnane 2 with various aldoximes (a-h) in presence of chloramine-T in refluxing ethanol. PMID- 15894033 TI - Formation of 19-norsteroids by in situ demethylation of endogenous steroids in stored urine samples. AB - The formation of 19-norsteroids by demethylation of endogenous steroids in stored urine samples was observed. Suspicious urine samples (i.e. containing trace amounts of 19-norandrosterone and 19-noretiocholanolone) were selected and spiked with deuterated analogues of androsterone and etiocholanolone at concentrations corresponding to high endogenous levels (4 microg/mL). After incubation, respective 19-norsteroids (19-norandrosterone-d4 and 19-noretiocholanolone-d5) were identified in these samples by high-resolution mass spectrometry. The transformation of the 5 beta-isomer (etiocholanolone) yields about three-fold higher concentrations, compared to the 5 alpha-isomer. A significant temperature dependence was observed by comparison of reaction kinetics at room temperature (23+/-2 degrees C) and 37 degrees C. Concentrations of 19-norandrosterone-d4 and 19-noretiocholanolone-d5, respectively, were 2.7 and 3.6 times higher at elevated temperature. The conversion of androsterone-d4 to 19-norandrosterone-d4 did not exceed a relative amount of 0.1%. Incubation of the urine samples with androsterone-d4-glucuronide led to the production of 19-norandrosterone-d4 glucuronoide. A partial stabilization was observed after addition of metabolic inhibitors (e.g. EDTA). The application of the incubation experiments described may contribute to the clarification of adverse analytical findings regarding low levels of 19-norsteroid metabolites. PMID- 15894034 TI - Inhibition of 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) activity of human lung microsomes by genistein, daidzein, coumestrol and C(18)-, C(19)- and C(21) hydroxysteroids and ketosteroids. AB - Epidemiologic data suggest a relationship between dietary intake of phytochemicals and a lower incidence of some cancers. Modulation of steroid hormone metabolism has been proposed as a basis for this effect. It has been shown that aromatase, 3beta-hydroxysteroid dehydrogenase and 17beta hydroxysteroid dehydrogenase (17beta-HSD) are inhibited by the isoflavones, genistein and daidzein, and by coumestrol. In general, the extent of inhibition has been expressed in terms of IC50-values, which do not give information as to the pattern of inhibition, i.e., competitive, non-competitive, or mixed. Less is known of the effects of these compounds on 3alpha-HSD. The human lung is known to have a high level of 17beta-HSD and 3alpha-HSD activity. During the course of studies to characterize both activities in normal and inflamed lung and lung tumors we noted that 3alpha-HSD activity with 5alpha-DHT of microsomes from normal, adult lung was particularly susceptible to inhibition by coumestrol. To clarify the pattern of inhibition, the inhibition constants Ki and K'i were evaluated from plots of 1/v versus [I] and [S]/v versus [I]. Genistein, daidzein and coumestrol gave mixed inhibition patterns versus both 5alpha-DHT and NADH. In contrast, 5alpha-androstane-3,17-dione and 5alpha-pregnane-3,20-dione were competitive with 5alpha-DHT. NAD inhibited competitively with NADH. Our findings demonstrate that phytochemicals have the potential to inhibit 5alpha-DHT metabolism and thereby affect the androgen status of the human lung. The observation of a mixed inhibition pattern suggests these compounds bind to more than one form of the enzyme within the catalytic pathway. PMID- 15894035 TI - Altered profiles of serum neuroactive steroids in premenopausal women treated for alcohol addiction. AB - Long-term alcohol consumption results in menstrual irregularities due to the inhibition of progesterone secretion. Some progesterone metabolites, including three pregnanolone isomers (PI), abate, while pregnenolone sulfate (PregS) and dehydroepiandrosterone sulfate (DHEAS) increase, alcohol tolerance. The rationale of this study was to evaluate how the neuroactive steroids reflect the impaired progesterone formation in premenopausal women treated for alcohol addiction, and whether detoxification therapy could restore female reproductive functions and psychosomatic stability by reinstatement of the steroid biosynthesis. Accordingly, serum allopregnanolone (3alpha-hydroxy-5alpha-pregnan-20-one (P3alpha5alpha)), pregnanolone (P3alpha5beta), isopregnanolone (P3beta5alpha) and epipregnanolone (P3beta5beta), progesterone, PregS, pregnenolone, 17alpha-hydroxy pregnenolone (Preg17), 17alpha-hydroxy-progesterone (Prog17), DHEA, DHEAS, cortisol and estradiol were measured in 20 women during the therapy (start, 3 days, 14 days, 1 month, 4 months), and in 17 controls, using GC-MS or RIA and evaluated by age-adjusted ANCOVA with status and phase of the menstrual cycle (PMC) as factors, and status-PMC interaction. The patients exhibited depressed progesterone, Prog17, PI, and estradiol, a decreased progesterone/pregnenolone ratio, a decreased ratio of neuroinhibiting P3alpha5alpha to neuroactivating PregS, and an elevated PregS and PregS/pregnenolone ratio. The treatment mostly restored the indices. The reduction of neuroinhibiting pregnanolone isomers in the patients is primarily associated with the impairment in ovarian steroid biosynthesis. Nevertheless, changes in enzyme activities connected with the formation of PI and the influence of altered physiological requirements on the balance between endogenous neuroinhibiting and neuroactivating steroids are also likely. The reinstatement of serum estradiol, progesterone, and PI during the therapy demonstrates its favorable effect on both reproductive functions and the psychosomatic stability of the patients. PMID- 15894036 TI - Characterization of the pharmacokinetics of dioscin in rat. AB - Dioscin (diosgenyl 2,4-di-O-alpha-l-rhamnopyranosyl-beta-d-glucopyranoside) is an important constituent of some traditional Chinese medicines with several bioactivities. We have investigated the pharmacokinetics of dioscin in rat after intravenous and oral administrations. Compartmental methods were used to perform pharmacokinetic data analysis. The dose-dependent pharmacokinetics of dioscin was characterized after intravenous administrations (0.064, 0.16, 0.4 and 1.0mg/kg) to rats. There was significant decrease in clearance with increasing dose (4.67+/ 0.09 ml/min/kg (0.064 mg/kg) versus 3.49+/-0.23 ml/min/kg (1.0 mg/kg), P<0.05), and the plot of reciprocal clearance values versus the doses was linear (r=0.909, P<0.05). After an I.V. dose of 1mg/kg, simultaneous oral gavage of activated charcoal did not change the pharmacokinetic parameters indicating enterohepatic recycling of dioscin is not important in rat. The absolute oral bioavailability was very low (0.2%). In tissue distribution and bile excretion studies after I.V. and oral administrations, dioscin was shown to undergo a prolonged absorption from the intestinal tract and slow elimination from organs, and only a small amount of drug was recovered in bile. The cumulative amounts of dioscin in feces and urine indicated that the parent drug is mainly excreted in the feces. PMID- 15894037 TI - Selective acylation of cholic acid derivatives with multiple methacrylate groups. AB - Bile acids in the family of steroid compounds can be chemically modified for biochemical and other applications. Derivatives of cholic acid with multiple methacrylate groups can be prepared by the use of methacrylic acid, methacryloyl chloride and methacryloyl anhydride as the acylating agents. The hydroxyl groups of cholic acid methyl ester and cholic acid ethylene glycol ester have been selectively acylated by changing the acylating agents and the number of substitutions have been varied by changing the amount of the acylating agents used. In the acylation reactions with methacryloyl chloride, the reactivity of secondary hydroxyl groups on the steroid skeleton of cholic acid derivatives follows the order of C3>C12>C7. PMID- 15894038 TI - Glycyrrhetinic acid, the active principle of licorice, can reduce the thickness of subcutaneous thigh fat through topical application. AB - Cortisol is involved in the distribution and deposition of fat, and its action is regulated by the activity of 11beta-hydroxysteroid dehydrogenase. Glycyrrhetinic acid, the active principle of licorice root, blocks 11beta-hydroxysteroid dehydrogenase type 1, thus reducing the availability of cortisol at the level of adipocytes. We evaluated the effect of topical application of a cream containing glycyrrhetinic acid in the thickness of fat at the level of the thigh. Eighteen healthy women (age range 20-33 years) with normal BMI were randomly allocated to treatment, at the level of the dominant thigh, with a cream containing 2.5% glycyrrhetinic acid (n=9) or with a placebo cream containing the excipients alone (n=9). Before and after 1 month of treatment both the circumference and the thickness of the superficial fat layer of the thighs (by ultrasound analysis) were measured. The circumference and the thickness of the superficial fat layer were significantly reduced in comparison to the controlateral untreated thigh and to control subjects treated with the placebo cream. No changes were observed in blood pressure, plasma renin activity, plasma aldosterone or cortisol. The effect of glycyrrhetinic acid on the thickness of subcutaneous fat was likely related to a block of 11beta-hydroxysteroid dehydrogenase type 1 at the level of fat cells; therefore, glycyrrhetinic acid could be effectively used in the reduction of unwanted local fat accumulation. PMID- 15894039 TI - Oxidative chemistry of 2-nitro and 4-nitroestradiol: Dichotomous behavior of radical intermediates and novel potential routes for oxyfunctionalization and B ring fission of steroidal scaffolds. AB - Oxidation of 4-nitro-17beta-estradiol (1) with the peroxidase/H(2)O(2) system gave the symmetric C(2)-linked dimer (3) through phenoxy radical coupling. Similar oxidation of 2-nitro-17beta-estradiol (2), in which the nitro group is coplanar with the aromatic ring, yielded 9alpha- and 9beta-hydroxy-2-nitro-17beta estradiol (4a,b), (17beta)-2-nitroestra-1(10),2,4,9(11)-tetraene-3,17-diol (5), and (12alpha,17beta)-2-nitroestra-1(10),2,4,9(11)-tetraene-3,12,17-triol (6). With higher concentrations of H(2)O(2), the novel secoestra-1(10),2,4-trien-9-one derivative 7 was obtained from 2. Theoretical calculations suggested that the peculiar behavior of 2 may be due to the generation of a relatively stable radical intermediate at C(9), which would then be converted to the reactive quinone methide 8. The chemistry described in this paper appears to be an intriguing example of control of the site of substitution over evolution of phenoxy radicals, and opens new vistas toward selective oxyfunctionalization of the estrane skeleton. PMID- 15894040 TI - Preparation of (25R)- and (25S)-26-functionalized steroids as tools for biosynthetic studies of cholic acids. AB - A new synthesis of both epimeric forms of 26-cholestanoic acids and 26-alcohols containing a 3beta-hydroxy-Delta(5)- or a Delta(4)-3-keto-functionality in ring A is described starting from stigmasterol or (20S)-3beta-acetoxy-pregn-5-en-20 carboxylic acid. The obtained compounds are useful as standards for studies of cholic acids. Construction of the side chain was achieved by linkage of steroidal 23-iodides to sulfones prepared from (2R)- and (2S)-3-hydroxy-2 methylpropanoates. Oxidation of intermediate 26-alcohols into the corresponding carboxylic acids ensuring preservation of stereochemistry at C-25 and functional groups in the cyclic part was achieved with sodium chlorite catalyzed by TEMPO and bleach. PMID- 15894041 TI - Uncertainty assessment of spatial patterns of soil organic carbon density using sequential indicator simulation, a case study of Hebei province, China. AB - The spatial patterns of soil organic carbon (SOC) are closely related to the global climate change. In quantifying the spatial patterns of SOC density, the concept of uncertainty of the SOC density values at unsampled locations is particularly important because such uncertainty can be propagated into the subsequent global climate change modelling and has fundamental impacts on the ultimate results of the model. A total of 361 SOC density data of topsoil (0-20 cm) in Hebei province and sequential indicator simulation (SIS) were applied to perform a conditional stochastic simulation in this study to quantitatively assess the uncertainty of mapping SOC density. The results showed that a great variation exists in the SOC density data. The conditional variance of 500 realizations generated by SIS was larger in mountainous areas of the study area where the SOC density fluctuated the most, and the uncertainty was smaller on the plain area where SOC density was consistently small. Realizations generated by SIS can represent the possible spatial patterns of SOC density without smoothing effect. A set of realizations can be used to explore all possible spatial patterns of SOC density and provide a visual and quantitative measure of the spatial uncertainty of mapping SOC density. With a given threshold of SOC density, SIS can quantitatively assess both local uncertainty and spatial uncertainty of SOC density that is greater the threshold. PMID- 15894042 TI - Composition of wet and bulk deposition in Erzurum, Turkey. AB - Seasonal variations in the chemical characteristics of wet and bulk deposition samples collected in Erzurum were investigated for the period March 2002-January 2003. Major cations (Ca2+, K+, Mg2+) and major anions (SO4(2-),NO3-) were determined in bulk and wet deposition samples; pH was also measured in wet deposition. The average pH of the wet deposition at Erzurum was 6.6 due to extensive neutralization of the acidity. A strong relationship between pH and SO4(2-) concentrations was observed in all seasons; however, only a weak relationship was found between pH and NO3-. On a seasonal basis, the correlation between Ca2+ and SO4(2-) concentrations was stronger in winter than in summer. Seasonal variations of ions were examined in both wet and bulk deposition samples. Although maximum concentrations of anions generally occurred during winter and spring, cation concentrations peaked in summer for both wet and bulk deposition. Results indicated that Ca2+ was the dominant cation and SO4(2-) the dominant anion in all deposition samples at Erzurum. Even though correlations among the crustal ions (calcium, magnesium and potassium) were high, the relationship between anthropogenic ions (sulfate and nitrate) was less clear in bulk deposition. PMID- 15894043 TI - Mitigation of methane and nitrous oxide emissions from drained irrigated rice fields. AB - One of the important cultural practices that affect methane and nitrous oxide emissions from tropical rice plantations is the water drainage system. While drainage can reduce methane emissions, it can also increase nitrous oxide emissions, as well as reduce yields. In this experiment, four different water drainage systems were compared in a rice field in central Thailand including: (1) continuous flooding, (2) mid-season drainage, (3) multiple drainage and (4) a local method (drainage was done according to local cultural practice) in order to find a system of drainage that would optimize yields while simultaneously limiting methane and nitrous oxide emissions. Methane and nitrous oxide emission were observed and compared with rice yield and physical changes of rice plants. It was found that drainage during the flowering period could reduce methane emission. Interestingly, nitrous oxide emission was related to number of drain days rather than the frequency of draining. Fewer drain days can help reduce nitrous oxide emission. The mid-season drainage and the multiple drainage, with 6.9% and 11.4% reduction in rice yield, respectively, had an average methane emission per crop 27% and 35% lower when compared to the local method. Draining with fewer drain days during the flowering period was recommended as a compromise between emissions and yield. The field drainage can be used as an option to reduce methane and nitrous oxide emissions from rice fields with acceptable yield reduction. Mid-season drainage during the rice flowering period, with a shortened drainage period (3 days), is suggested as a compromise between the need to reduce global warming and current socio-economic realities. PMID- 15894044 TI - Characteristics of roadside air pollution in Korean metropolitan city (Daegu) over last 5 to 6 years: temporal variations, standard exceedances, and dependence on meteorological conditions. AB - The present study performed a roadside data analysis to provide baseline data for exploring associations between environmental exposure to four gaseous pollutants and health effects on residents living near roadways. The yearly roadside concentrations of CO and SO2 showed a well-defined decreasing trend, whereas those of NO2 and O3 exhibited the reverse trend. In most cases, the diurnal trends of the roadside concentrations were well-defined for all seasons, plus the daytime concentrations were higher than the nighttime concentrations. In contrast to the other target pollutants, the daytime O3 concentrations observed at the roadside sites were lower than those observed at the residential site, likely due to high-levels of fresh NO from traffic emissions that rapidly react with O3, thereby reducing the O3 roadside level. The Sunday roadside concentrations of CO, NO2, and SO2 were similar to or somewhat lower than the weekday concentrations. Conversely, for O3 the Sunday roadside concentrations were similar to or somewhat higher than the weekday concentrations. The higher O3 concentrations on Sunday may be due to the reduced titration from a decrease in NOx emissions under VOC limited conditions (low VOC/NOx conditions). The monthly averages of O3 concentrations exhibited the reverse seasonal variation to the other target compounds, with peak O3 concentrations between April and June, and the second peak between August and October. It is also suggested that for O3, the 8-h standard is more stringent than the 1-h standard, while for NO2 and SO2, the 1-h standard is more stringent than the 24-h standard. The multiple regression equations obtained from the relationship between the concentrations and five meteorological parameters indicated that the number and type of meteorological variables in the equations varied according to the pollutant, monitoring station, or season. PMID- 15894045 TI - Baseline study of methane emission from open digesting tanks of palm oil mill effluent treatment. AB - Anthropogenic release of greenhouse gases, especially CO2 and CH4 has been recognized as one of the main causes of global warming. Several measures under the Kyoto Protocol 1997 have been drawn up to reduce the greenhouse gases emission. One of the measures is Clean Development Mechanisms (CDM) that was created to enable developed countries to cooperate with developing countries in emission reduction activities. In Malaysia, palm oil industry particularly from palm oil mill effluent (POME) anaerobic treatment has been identified as an important source of CH4. However, there is no study to quantify the actual CH4 emission from the commercial scale wastewater treatment facility. Hence, this paper shall address the CH4 emission from the open digesting tanks in Felda Serting Hilir Palm Oil Mill. CH4 emission pattern was recorded for 52 weeks from 3600 m3 open digesting tanks. The findings indicated that the CH4 content was between 13.5% and 49.0% which was lower than the value of 65% reported earlier. The biogas flow rate ranged between 0.8l min(-1)m(-2) and 9.8l min(-1)m(-2). Total CH4 emission per open digesting tank was 518.9 kgday(-1). Relationships between CH4 emission and total carbon removal and POME discharged were also discussed. Fluctuation of biogas production was observed throughout the studies as a result of seasonal oil palm cropping, mill activities, variation of POME quality and quantity discharged from the mill. Thus only through long-term field measurement CH4 emission can be accurately estimated. PMID- 15894046 TI - Methane fluxes from tundra soils and snowpack in the maritime Antarctic. AB - Methane fluxes were measured from three exposed tundra sites and four snowpack sites on the Fildes Peninsula in the maritime Antarctic in the summertime of 2002. The average fluxes at two normal tundra sites were -15.3 microgm(-2)h(-1) and -14.3 microgm(-2)h(-1), respectively. The fluxes from tundra site with fresh penguin dropping addition showed positive values with the average of 36.1 microgm(-2)h(-1), suggesting that the deposition of fresh droppings greatly enhanced CH4 emissions from the poor Antarctic tundra during penguin breeding periods. The summertime variation in CH4 flux was correlated with surface ground temperature and the precipitation. The correlation between the flux and PT0, which is the product of the precipitation and surface ground temperature, was quite strong. The diurnal cycle of CH4 flux from the tundra soils was not obtained due to local fluky weather conditions. The fluxes through four snowpack sites were also obtained by the vertical CH4 concentration gradient and their average fluxes were -46.5 microgm(-2)h(-1), -28.2 microgm(-2)h(-1), -46.4 microgm(-2)h(-1) and -17.9 microgm(-2)h(-1), respectively, indicating that tundra soils under snowpack also consume atmospheric CH4 in the maritime Antarctic; therefore these fluxes could constitute an important part of the annual CH4 budget for Antarctic tundra ecosystem. PMID- 15894047 TI - Application of micrometeorological approaches to measure methane exchange in a dry paddy field in the western coast of Korea. AB - The exchange processes of CH4 were investigated in a paddy field in the Hari area of Kang Hwa Island over an 8 day period in late April 2002. The quantification of CH4 fluxes was made under dry field condition of early spring by concurrently measuring its concentrations (at the two heights of 1 and 5 m) and the relevant micrometeorological parameters. To help elucidate the factors determining the mobilization characteristics of CH4, the results of our measurement data were examined using a number of approaches. The results of the trajectory analysis indicated that its concentration changed very sensitively with the influence of different source types, as seen from the air mass movement patterns. The concentrations and fluxes of methane, when examined over this short-term scale, showed moderately strong patterns across 24h period in which higher values tend to occur during morning or evening. The overall results of our field measurements suggest that CH4 exchange processes in the paddy area proceeded in a fairly complicated manner. The study area behaved as a net source of CH4 to the atmosphere with a net daily emission rate of 3.6 mgm(-2) despite the fact that downward deposition was observed more frequently than upward emission. PMID- 15894048 TI - A microcosm approach to assessing the effects of earthworm inoculation and oat cover cropping on CO2 fluxes and biological properties in an amended semiarid soil. AB - We designed a microcosm experiment to assess the influence of inoculation with Eisenia foetida earthworms and the establishment of an Avena sativa cover crop on biological (enzyme activities and labile carbon fractions) soil quality indicators in a soil treated with a composted organic residue, and to determine the contribution of these treatments to carbon dioxide emissions from the soil to the atmosphere of the microcosm. The microcosms were incubated for 53 days under 28 degrees C/18 degrees C day/night temperatures. The addition of earthworms and the planting of A. sativa increased dehydrogenase activity of compost amended soil by about 44% after 23 days of incubation. The metabolic potential, calculated as the ratio dehydrogenase activity/water soluble C, was higher in the compost amended soil planted with A. sativa. The highest total amount of CO2-C evolved occurred in the soil treated with composted residue and earthworms (about 40% of the total amount of CO2 evolved came from earthworm activity). The planting of A. sativa increased the decomposition rate constant of organic matter in the amended soil but decreased the potentially mineralizable C pool. In conclusion, the establishment of an A. sativa cover crop and the addition of E. foetida to a degraded agricultural soil treated with composted residue were effective treatments for improving the biological and biochemical quality and the metabolic potential of the soil. PMID- 15894049 TI - Impacts of some meteorological parameters on SO2 and TSP concentrations in Erzurum, Turkey. AB - The air pollution is the one of the most important environmental problems in Erzurum, situated in the eastern of Turkey, during winter periods. The unfavorable climate as well as the city's topography, and inappropriate urbanization cause serious air pollution problems. The air pollutant concentrations in a city have a close relationship with its meteorological parameters. In the present study, the relationship between daily average total suspended particulate (TSP) and sulphur dioxide (SO2) concentrations with meteorological factors, such as wind speed, temperature, relative humidity, pressure and precipitation, in 1995-2002 winter seasons was statistically analyzed using the stepwise multiple linear regression analysis. According to the results obtained through analysis, higher TSP and SO2 concentrations are strongly related to colder temperatures, lower wind speed, higher pressure system and weakly lower precipitation and higher relative humidity. The statistical models of SO2 and TSP including meteorological parameters gave R2 of 0.74 and 0.88, respectively. Furthermore, the correlation between the previous day's SO2, TSP concentrations and actual concentrations of these pollutants on that day was investigated and found as 0.84 and 0.53, respectively. In order to develop this model, previous day's SO2 and TSP concentrations were added to the equations. The new model for SO2 enhanced considerably (R2 = 0.92), but for TSP new model was not enhanced (R2 = 0.89). PMID- 15894050 TI - Emission of volatile organic compounds (VOC) from tropical plant species in India. AB - Foliar emission of volatile organic compounds (VOC) from common Indian plant species was measured. Dynamic flow enclosure technique was used and the gas samples were collected onto Tenax-GC/Carboseive cartridges. The Tenax GC/Carboseive cartridges were attached to the thermal disorber sample injection system and the gas sample was analysed using gas chromatography (GC) with flame ionisation detection (FID). Fifty-one local plant species were screened, out of which 36 species were found to emit VOC (4 high emitter; 28 moderate emitter; and 4 low-emitter), while in the remaining 15 species no VOC emission was detected or the levels of emission were below detection limit (BDL). VOC emission was found to vary from one species to another. There was a marked seasonal and diurnal variation in VOC emission. The minimum and maximum VOC emission values were < 0.1 and 87 microgg(-1) dry leaf h(-1) in Ficus infectoria and Lantana camara respectively. Out of the 51 plant species studied, 13 species are reported here for the first time. Among the nine tree species (which were selected for detailed study), the highest average hourly emission (9.69+/-8.39 microgg(-1) dry leaf) was observed in Eucalyptus species and the minimum in Syzygium jambolanum (1.89+/ 2.48 microgg(-1) dry leaf). An attempt has been made to compare VOC emission from different plant species between present study and the literature (tropical and other regions). PMID- 15894051 TI - Hydrocarbon and elemental carbon signatures in a tropical wetland: biogeochemical evidence of forest fire and vegetation changes. AB - Evidence of changing vegetation in the tropical wetland (Rawa Danau, west Java, Indonesia) over the past 7428 years is illustrated by elemental (soot) carbon (EC) and n-alkane composition of sedimentary geolipids. In this study, vegetation changes and relevant controlling factors (e.g. forest fire and climate change) were documented on a decadal to centennial scale. The n-alkane composition that changes with depth might record changes in sources of organic matter (OM) in the wetland. The presence of EC (0.01-0.24% of organic carbon: OC) during late (0 1700 cal. year BP) and mid (3500-4500 cal. year BP) Holocene (at depths 0-50 cm, and 160-210 cm) indicated that large-scale forest fires severely affected the tropical vegetation. The hydrocarbon indices (CPI: carbon preference index, MCN: mean carbon number, and HVI: hydrocarbon vegetation index) significantly correlated with one another while a comparison of EC profile with the profiles of hydrocarbon indices indicated that n-alkane composition of the geolipid in lake sediment could record signatures of changes in catchment vegetation. Forest fire and vegetation changes might be related to regional climatic shifts relating to ENSO activity as well as being influenced by human influences. PMID- 15894052 TI - Nitrous oxide emissions from tundra soil and snowpack in the maritime Antarctic. AB - The nitrous oxide emissions were measured at three tundra sites and one snowpack on the Fildes Peninsula in the maritime Antarctic in the summertime of 2002. The average fluxes at two normal tundra sites were 1.1+/-2.2 and 0.6+/-1.7 microg N2O m(-2)h(-1), respectively. The average flux from tundra soil site with penguin dropping addition was 3.7+/-2.0 microg N2O m(-2)h(-1), 3-6 times those from the normal tundra soils, suggesting that the deposition of fresh droppings enhanced N2O emissions during penguin breeding period. The summer precipitation had an important effect on N2O emissions; the flux decreased when heavy precipitation occurred. The diurnal cycle of the N2O fluxes from Antarctic tundra soils was not obtained due to local fluky weather conditions. The N2O fluxes through four snowpack sites were obtained by the vertical N2O concentration gradient and their average fluxes were 0.94, 1.36, 0.81 and 0.85 microg N2O m(-2)h(-1), respectively. The tundra soils under snowpack emitted N2O in the maritime Antarctic and increased local atmospheric N2O concentrations; therefore these fluxes could constitute an important part of the annual N2O budget for Antarctic tundra ecosystem. PMID- 15894053 TI - Methane emission characteristics and its relations with plant and soil parameters under irrigated rice ecosystem of northeast India. AB - Methane flux from rice varieties grown under two identical soils of Assam were monitored. In the first experiment, variety Jaya and GRT was grown in sandy loam soil of Lower Brahmaputra Valley Zone of Assam and the second experiment was conducted with variety Jyotiprasad and Bishnuprasad in sandy to sandy loam soils of Upper Brahmaputra Valley Zones of Assam. Methane flux recorded from variety Jyotiprasad and GRT was higher compared to variety Bishnuprasad and Jaya. The seasonal integrated flux recorded was 10.76 gm(-2), 9.98 gm(-2), 9.74 gm(-2) and 11.31 gm(-2) for variety GRT, Jaya, Bishnuprasad and Jyotiprasad, respectively. All the varieties exhibited two methane peaks one at maximum tillering stage and other at panicle initiation stage of the crop. Crop growth parameters such as leaf number, number of tillers and leaf area index (LAI) showed strong positive relationship with total methane flux. In both the experiments it was calculated that CH4 emission was substantially influenced by crop phenology and growth. This study emphasise the relationship of different growth parameters with methane emission. PMID- 15894054 TI - Seasonal variations of monoterpene emissions from coniferous trees of different ages in Korea. AB - Seasonal variations of emission rates and compositions from coniferous species were measured under controlled conditions using a vegetation enclosure method. Total emission rates and compositions of monoterpene compounds from young and adult trees in different seasons were compared. It was found that the total emission rates and the components of monoterpene varied significantly with tree species, age, and season. Total emissions from C. japonica and P. koraiensis were higher for older trees than for younger trees; however, significantly higher emissions were found from younger trees for C. obtusa. Higher monoterpene emission rates from each plant were found in spring and summer compared with autumn and winter emissions. PMID- 15894055 TI - Biogeochemical distinction of methane releases from two Amazon hydroreservoirs. AB - Biogeochemical distinction of methane emissions to the atmosphere may essentially rely on the surface area and morphometry of Amazon hydroreservoirs. Tucurui (deep) and Samuel (shallow) reservoirs released in average 13.82+/-22.94 and 71.19+/-107.4 mg CH4 m(-2)d(-1), respectively. delta13C-CH4 values from the sediments to the atmosphere indicate that the deep reservoir has extended methanotrophic layer, oxidizing large quantities of light isotope methane coming from the sediments, while sediment-generated methane can easily evade the shallow reservoir. PMID- 15894056 TI - Cold season CH4, CO2 and N2O fluxes from freshwater marshes in northeast China. AB - Cold season (winter and thaw) CH4, CO2 and N2O fluxes from freshwater marshes (47 degrees 35'N, 133 degrees 31'E, Northeast China) were measured, using the static chamber method. The mean CH4 and CO2 fluxes from Carex lasiocarpa (Cl) were 0.5+/ 0.19 and 6.23+/-1.36 mgCm(-2)h(-1), respectively, and those from Deyeuxia angustifoli (Da) were 0.18+/-0.15 and 5.22+/-2.48 mgCm(-2)h(-1), respectively in winter. There was no significant difference between Cl and Da (p > 0.05). The contributions of winter CH4 fluxes were about 5.5% and 3% in the Cl and Da, respectively. Marshes are an important potential N2O sink in winter season in northeast China. During thaw, the CH4 and CO2 emissions rapidly increased, 4.5-6 times of winter emissions. Wetland became a source of N2O. Cold season gases flux from northern wetlands play an important role in the seasonal gas exchange. PMID- 15894057 TI - Withdrawal, tolerance, and sensitization to dopamine mediated interoceptive cues in rats trained on a three-lever drug-discrimination task. AB - In the present experiment rats were trained on a three-lever, drug-discrimination task to discriminate the cues associated with 0.30 mg/kg of the indirect dopamine (DA) agonist, amphetamine (AMPH), saline (SAL), and 0.03 mg/kg of the DA, D2 receptor antagonist, haloperidol (HAL). Choice behavior determined from tests on 0.30 and 0.15 mg/kg AMPH, SAL 0.03 and 0.015 mg/kg HAL provided a behavioral baseline presumed to represent changes along a continuum of DA mediated, interoceptive cues. Results from separate groups tested on 0.30 and 0.15 mg/kg AMPH, SAL, 0.03 and 0.015 mg/kg HAL, 24 h post-treatment with an acute 7.5 mg/kg dose of AMPH, showed rapid tolerance and withdrawal to the AMPH cue and sensitization to the HAL cue. The same tests 24 h following treatment with 1.0 mg/kg HAL showed rapid tolerance to the HAL cue, sensitization to the AMPH cue, but not AMPH-like withdrawal cues. Analysis of the results showed that tolerance to the AMPH and HAL cues reflected neuroadaptive baseline shifts and not weaker cue properties. These findings are consistent with predictions from opponent process theory of motivation and provide an animal model to study the motivational consequences that aversive symptoms of AMPH withdrawal such as dysphoria and anhedonia can have on drug-taking behavior. PMID- 15894058 TI - Exacerbation of harmaline-induced tremor by imipramine. AB - Imipramine is a well-established tricyclic antidepressant which was first approved for the treatment of depression in the late fifties. Antidepressant effect of imipramine is attributed to inhibition of serotonin (5HT) and noradrenaline (NA) reuptake in brain. These monoamines have been implicated in a variety of neurological disorders including tremor. In the present investigation attempt was made to study the effect of imipramine on harmaline-induced tremor in rats. Male Sprague Dawley rats weighing 115+/-2.5 g were given harmaline (10 mg/kg, i.p.) alone or along with imipramine (30 min before harmaline) in doses of 60 and 90 mg/kg respectively. The latency of onset, intensity and duration of tremor and EMG were recorded. To substantiate the role of 5HT in aetiopathology of tremor the above experiment was repeated in the rats pretreated with P chlorophenylalanine (PCPA), a potent 5HT depleter. The levels of 5HT and 5 hydroxyindole acetic acid (5HIAA) in the brain stem were measured using high performance liquid chromatography. Imipramine dose-dependently exacerbated the duration, intensity and amplitude of EMG following harmaline-induced tremor. Imipramine treatment further decreased harmaline-induced 5HT turnover in the brain stem. However, this was statistically insignificant. Depletion of 5HT produced a significant reduction in the intensity and duration of harmaline induced tremor. In conclusion, this study suggests that imipramine exacerbates harmaline-induced tremor. Clinical use of imipramine for the treatment of depression in patients who also suffer from tremors may require a close monitoring. PMID- 15894059 TI - Behavioral, biochemical and histological studies in a model of pilocarpine induced spontaneous recurrent seizures. AB - Although the presence of profound cognitive disturbances in lithium-pilocarpine induced spontaneous recurrent seizures (SRS) has been well documented, much less is known about changes in emotional behavior, in this model of temporal lobe epilepsy. To that end, a lithium-pilocarpine model of SRS was used to evaluate behavior of experimental animals (SRS, non-SRS and saline-treated rats) in different tests of anxiety (open field test, fear conditioning freezing and footshock-induced ultrasonic vocalization). Flinch-jump test, allowing determination of pain threshold, was employed to confirm specificity of data from anxiety tests. Moreover, neurotransmitters' (dopamine, serotonin and their metabolites) concentration was measured in selected brain structures involved in emotional and motor processing (hippocampus, frontal cortex and striatum). Finally, different brain structures were examined histologically in order to determine structures likely to be involved in behavioral changes. It was found that SRS rats, tested in a seizure free period, revealed an increase in motor activity, and a decrease in fear-related reactions (a freezing response to the aversively conditioned context, and a spontaneous, emotion-related ultrasonic vocalization). No changes in the pain threshold were present. The activity of dopamine and serotonin systems in examined brain structures remained unchanged. The neuropathological effects were widespread and involved a loss of neurons, proliferation of astroglial cells and the presence of activated ramified and ameboid microglial cells in the hippocampus proper, piriform cortex, amygdala and lateral posterior thalamic nuclei. The obtained results suggest a prevalence of disinhibitory effects on behavior in SRS rats, as shown by the results of contextual fear and aversive vocalization tests (i.e. a release of rat behavior controlled by fear). It is conceivable that the lesions to the limbic structures involved in the origin of emotions; the hippocampus, amygdala, and piriform cortex, may underlie changes in anxiety reactions in SRS rats. These results indicate that lithium-pilocarpine-induced SRS are also accompanied by profound alteration of animal emotional behavior. PMID- 15894060 TI - Autonomic efferents affect intake of imbalanced amino acid diets by rats. AB - An anorectic response occurs following ingestion of imbalanced amino acid (IMB) diets. There are three phases to this response: 1, recognition of the IMB diet; 2, conditioned development of an aversion to the IMB diet; and 3, adaptation. Blockade of peripheral serotonin-3 (5-HT3) receptors or vagotomy attenuates Phase 2 of the anorectic response. We investigated whether sympathetic efferents interact with the ventral gastric branch (VGB), by cutting it (X), or with the 5 HT3 receptor in these responses. First, VGBX and sham-operated (SHAM) groups were injected with vehicle or phenoxybenzamine (alpha-blocker), or nadolol (beta blocker) before introducing the IMB diet. At 3 h suppression of the IMB diet ingestion was unchanged, showing no sympathetic efferent effect on Phase 1. Intake of the IMB diet increased 12-24 h later only in the SHAM+phenoxybenzamine group, so the VGB was necessary for alpha-blockade to enhance IMB diet intake during Phase 2 or possibly Phase 3. On days 2-5, intakes by the SHAM+phenoxybenzamine, VGBX+phenoxybenzamine and VGBX+nadolol groups were elevated. Therefore, alpha-blockade enhanced adaptation alone, but VGBX was necessary for beta-receptor blockade to augment Phase 3 adaptation. Both sympathetic efferents and the VGB are involved in Phases 2-3. Second, rats received vehicle or nadolol or scopolamine (nonselective muscarinic blocker) or pirenzepine (muscarinic M-1 receptor blocker),w+/-tropisetron (5-HT3 blocker). Pirenzepine attenuated the tropisetron effect between 6-9 h, but then pirenzepine and nadolol enhanced the tropisetron effect between 9-12 h. Scopolamine attenuated the tropisetron effect between 9-12 h. While neither experiment showed effects during the recognition phase, the autonomic and serotonergic systems interact in the learned and adaptive responses to IMB diets. PMID- 15894061 TI - Effects of nitric oxide synthase inhibition on spatial discrimination learning and central DA2 and mACh receptors. AB - Cholinergic and dopaminergic systems are involved in spatial memory and are modulated by nitric oxide (NO); NO has well documented effects on place learning in rodents. The aim of the present study was to investigate the effect of NOS inhibition on place learning in the water maze and to evaluate the relationships between NOS inhibition, learning performance, dopamine (DA) D2 and muscarinic acetylcholine (mACh) receptors. Male Sprague-Dawley rats received the NOS inhibitor Nomega-Nitro-l-Arginine (l-NA), or saline and were trained in the water maze. Rats that were not trained, but received the same treatments were also included. Following treatments with or without water maze training, [3H]-QNB and [3H]-spiperone binding in cortex, striatum and hippocampus were determined to assess the effects of NOS inhibition and/or learning on DA D2 and mACh receptor regulation. The overall results of the present study showed that: (1) NOS inhibition impairs performance in the MWM; (2) NOS inhibition does not affect specific binding to DA D2 (striatum and hippocampus) and mACh (cortex and hippocampus) receptors; (3) MWM training lowers D2 and mACh receptor binding in cortical regions. PMID- 15894062 TI - The effects of chronic food restriction on hypothalamic-pituitary-adrenal activity depend on morning versus evening availability of food. AB - Partial food restriction (FR) protocols have been used not only to study behavioral and physiological consequences of decrease food intake, but as a necessary treatment of the animals in some operant learning tasks. It is well established in rodents that restricting food availability to a few hours in the morning causes an alteration of the daily rhythm of corticosterone, thus making it difficult to evaluate whether or not such treatments are stressful. In the present experiment adult male Sprague-Dawley rats were subjected to two different FR schedules: food availability after 1100 h (LFR) or after 1900 h (DFR). After 14 days, animals from both groups, together with corresponding controls, were killed under resting conditions, either in the morning or in the evening, just before daily access to food in FR rats. Both FR schedules reduced body weight gain to the same extent, but their impact on the hypothalamic-pituitary-adrenal (HPA) axis was different: DFR increased relative, but not absolute, adrenal weight and morning and evening levels of corticosterone, whereas LFR increased both absolute and relative adrenal weights and increased morning corticosterone levels to a greater extent than DFR rats. Neither serum ACTH nor corticotropin releasing factor (CRF) mRNA levels in the paraventricular nucleus of the hypothalamus were altered by DFR or LFR protocols, suggesting that factors other than CRF and ACTH are involved in the control of adrenocortical secretion under FR. It appears that LFR caused more alterations in the HPA axis than DFR and, therefore, the latter FR schedule should be used in those protocols necessarily involving partial FR. PMID- 15894063 TI - Amino acid levels in some lethargic mouse brain areas before and after pentylenetetrazole kindling. AB - Genetic animal models have contributed significantly to our understanding of epilepsy causes. Lethargic mice are considered a valid model of absence epilepsy, which have been shown to possess behavioral, electrographic and pharmacological profiles similar to those of humans with absence epilepsies. Single gene mutations that comprise the beta4 subunit of voltage-sensitive Ca2+ channels underlie the spontaneous discharges of the absence, non-convulsive seizures of lethargic mice. There are no available data concerning how the mutant channels actually behave at terminals in response to chemical activation by subconvulsant stimulation with pentylenetetrazole. In this study, we found no significant difference in the convulsive dose 50 between lethargic and control mice. Lethargic mice showed a more rapid development of kindling to pentylenetetrazole than control animals. No significant differences were observed between the groups of mice rechallenged with pentylenetetrazole 30 or 60 days after the end of the chronic treatment. Marked differences in brain amino acid levels were found between the two strains of mice in basal conditions and after kindling. In conclusion, our results indicate that lethargic mice show a range of biochemical and behavioral changes, correlated in particular with a higher susceptibility to develop kindled seizures. PMID- 15894064 TI - Neonatal ethanol and nicotine exposure causes locomotor activity changes in preweanling animals. AB - Sprague-Dawley rats were used to investigate the effects of neonatal ethanol (ETOH) and nicotine (NIC) exposure on activity levels in preweanling offspring. Male and female pups received daily oral intubations of ethanol ((ETOH) 5 g/kg/day), nicotine ((NIC) 12 mg/kg/day), ethanol and nicotine ((ETOH+NIC) 5 g/kg/day+12 mg/kg/day) or isocaloric maltose (control) on either postnatal days (PND) 1-7 or PND 8-14. A non-treated control group was also included. Peak blood ethanol concentrations (BECs) measured in a separate subset of animals ranged from 167 and 344 mg/dl depending upon neonatal treatment and period of exposure. Subjects were tested in an open field apparatus on PND 19-21. Animals exposed to ETOH or ETOH+NIC on PND 1-7 were hyperactive relative to the other treatment groups. In contrast, animals exposed to NIC or ETOH+NIC during PND 8-14 were hypoactive relative to other treatment groups. Males appeared more sensitive than females on measures of anxiety (distance traveled in the center of the open field) but this also varied dependent on neonatal treatment and period of exposure. These findings suggest that the third trimester is a critical period for ETOH and NIC effects on offspring activity although the pattern of effects on activity are different depending on when drug exposure occurred during the neonatal period. PMID- 15894065 TI - Effects of monoamine reuptake inhibitors on wet-dog shakes mediated by 5-HT2A receptors in ACTH-treated rats. AB - We examined the influence of imipramine, a serotonin (5-HT) and noradrenaline (NA) reuptake inhibitor, desipramine, a NA reuptake inhibitor, bupropion, a dopamine reuptake inhibitor, fluvoxamine, a selective 5-HT reuptake inhibitor, and mazindol, a catecholamine reuptake inhibitor, on a 5-HT2A receptor-mediated behavior, (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI)-induced wet dog shakes, in naive and adrenocorticotropic hormone (ACTH)-treated rats. Chronic administration of imipramine, desipramine and mazindol suppressed the number of wet-dog shakes in naive rats. Chronic ACTH (100 microg/rat, s.c.) treatment increased the number. Chronic administration of imipramine did not decrease the number of wet-dog shakes in ACTH-treated rats. On the other hand, desipramine and mazindol inhibited the increase in wet-dog shakes in ACTH-treated rats. Fluvoxamine and bupropion did not have any effect on the (+/-)-DOI-induced response in naive and ACTH-treated rats. NA reuptake inhibitors may improve the hyperfunction of 5-HT2A receptors induced by chronic ACTH treatment. PMID- 15894066 TI - Acute delta9-tetrahydrocannabinol exposure facilitates quinpirole-induced hyperlocomotion. AB - The endogenous cannabinoid system works as a feedback signal controlling dopamine induced facilitation of motor behaviors. The present study explored whether a single acute stimulation of CB1 cannabinoid receptors with (-)-Delta9 tetrahydrocannabinol (THC, 5 mg kg(-1) i.p.) results in modifications in the sensitivity to the acute behavioral effects of the dopamine D2/D3 receptor agonist quinpirole (0.025, 0.25 and 1 mg kg(-1), s.c.) 24 h after THC administration. Cannabinoid pretreatment increased the sensitivity to quinpirole induced hyperlocomotion 24 h after its administration. The data indicated that THC induced a desensitization of cannabinoid receptors, as revealed by a reduction in CB1 receptor-agonist induced GTP-gamma-S incorporation in striatal membranes. These results might be relevant for understanding the effect of cannabinoid exposure in dopamine-related neuropsychiatric disorders. PMID- 15894067 TI - Behavioral effects of the novel cannabinoid full agonist AM 411. AB - AM 411 ((-)-1-adamantyl-Delta8-tetrahydrocannabinol) is a novel full agonist at cannabinoid CB1 receptors. The present studies were conducted to provide behavioral characterization of this compound in rats. It was hypothesized that AM 411 should produce behavioral effects similar to known cannabinoid agonists, and that these effects should be inhibited by co-treatment with a CB1 antagonist. In Experiments 1 and 2, AM 411 dose-dependently produced behaviors consistent with CB1 agonism, including analgesia, hypothermia, catalepsy and reductions in locomotion, which were blocked by a CB1-selective antagonist. In Experiment 3, AM 411 produced a dose-dependent suppression of lever-pressing on a fixed-ratio 5 (FR5) schedule, a task known to be sensitive to administration of CB1 agonists. Detailed analysis of the temporal patterns of operant responding showed that AM 411 altered the distribution of interresponse times. Experiment 4 showed that AM 411 decreased relative interior activity in the open field, which is suggestive of an anxiogenic effect. It is concluded that AM 411 produces CB1 agonist-like behavior with potency between that of WIN 55,212-2 and AM 356. AM 411 could be a useful tool for understanding the behavioral and neural effects of CB1 receptor stimulation. PMID- 15894068 TI - Analgesic effects of loperamide in bone cancer pain in mice. AB - The intratibial inoculation of NCTC 2472 cells induces an osteosarcoma in C3H/HeJ mice. These mice show thermal hyperalgesic responses which may be blocked by the local administration of opiates over the tibial tumoral mass (Menendez L, Lastra A, Hidalgo A, Meana A, Garcia E, Baamonde A. Peripheral opioids act as analgesics in bone cancer pain in mice. NeuroReport 2003b; 14:867-9). The aim of this report was to characterize the analgesic responses obtained by activating peripheral opioid receptors in bone cancer pain. Here, we initially describe that this osteosarcoma induces mechanical as well as thermal hyperalgesia. Loperamide, an opioid agonist unable to cross the blood-brain barrier, inhibits both thermal and mechanical hyperalgesia when s.c. injected, locally over the tibial tumoral mass (7.5-75 microg) or distantly, under the fur of the neck (4 mg/kg). These analgesic effects seem peripherally mediated since they are reverted by the administration of naloxone methiodide (10 mg/kg) and because the withdrawal latencies of the contralateral, non-affected, paws remain unaltered. Furthermore, only cyprodime (1 mg/kg) but not naltrindole (0.1 mg/kg) or nor-binaltorphimine (10 mg/kg) blocked these effects, showing the involvement of gamma-opioid receptors in the peripheral analgesia induced by loperamide on thermal and mechanical hyperalgesia. The advantages of using peripheral acting opiates -- devoid of central colateral effects -- for the treatment of cancer related pain are suggested. PMID- 15894069 TI - Pharmacological modulation of repeated ethanol withdrawal-induced anxiety-like behavior differs in alcohol-preferring P and Sprague-Dawley rats. AB - Previous work with Sprague-Dawley (SD) rats indicated that subjecting these rats to multiple episodes of ethanol diet could provoke anxiety-like responses. Because alcohol-preferring P rats have been reported to have neurochemical alterations in many systems shown to modulate anxiety-like responses, P rats were compared to SD rats. Rats were subjected to one or three cycles of 5 days' exposure to 4.5% or 7% ethanol diet to assess anxiety-like behavior. The social interaction test was conducted 5 h after ethanol was removed. Other groups of P and SD rats were injected with flumazenil (5 mg/kg), a benzodiazepine (BZD) receptor antagonist, CP-154,526 (10 mg/kg), CRF1 receptor antagonist, SB243,213, a 5-HT2C receptor inverse agonist, or vehicle during the 1st and 2nd withdrawals but not the third. After a single 5-day cycle of ethanol exposure, SD rats did not exhibit a change in social interaction, but P rats exhibited a decrease after exposure to the 7% ethanol. Both strains of rats exhibited anxiety-like behavior following three cycles of exposure to ethanol and the concentration of ethanol in the diet did not influence the response. It was confirmed that flumazenil, CP 154,523, and SB243,213 had prophylactic effects on anxiety-like behavior in the SD rats. Neither flumazenil nor SB243,213 was as effective in the P rats, while the CRF1 receptor antagonist completely counteracted the reduced social interaction in repeatedly withdrawn P rats. A small study showed that buspirone, a 5-HT1A agonist, also had prophylactic effects in P rats. These findings show that alcohol-preferring P rats exhibit anxiety-like behavior more readily following exposure to ethanol-containing diets and that this behavior is counteracted more readily by pretreatment with a CRF1 receptor antagonist than with BZD or 5-HT2C receptor antagonists. PMID- 15894070 TI - The influence of dextromethorphan on morphine analgesia in Swiss Webster mice is sex-specific. AB - NMDA (N-methyl-d-aspartate) antagonists are known to enhance the analgesic effects of opioids. However, virtually, all studies of this phenomenon have been done using male subjects. Here, the noncompetitive NMDA receptor antagonist dextromethorphan (DEX) was tested over a range of doses (10-200 microg intracerebroventricularly [i.c.v.]) in male and female Swiss Webster mice in combination with 5 mg/kg intraperitoneal (i.p.) morphine. Males exhibited enhanced morphine analgesia following either 100 or 200 microg DEX, but there was no evidence of DEX-mediated potentiation in females at any dose. Instead, DEX attenuated morphine analgesia in females. We also evaluated the effect of 100 microg i.c.v. DEX with different doses of morphine (1, 5 and 10 mg/kg). Again, DEX significantly enhanced morphine analgesia in male mice and attenuated it in females. Next, ovariectomized (OVX) female mice were compared to males following 5 mg/kg i.p. morphine and 100 microg i.c.v. DEX. Male and OVX females exhibited equivalent maximal levels of analgesia following administration of DEX. Morphine analgesia was not enhanced by DEX in sham-treated females and OVX mice with estradiol treatment (5 microg i.p. once daily for 7 days) also did not show DEX enhancement. These experiments demonstrate that the ability of NMDA receptor antagonists to potentiate morphine analgesia is modulated by an estrogen sensitive mechanism and suggest that sex differences may play a critical role toward a more general understanding of the potentiation of opioid-induced analgesia through NMDA receptor antagonists. PMID- 15894071 TI - Extinction and reacquisition of a fear-motivated memory require activity of the Src family of tyrosine kinases in the CA1 region of the hippocampus. AB - Evidences indicate that extinction represents a NMDA receptor (NMDAr)-dependent learning rather than erasure of previously stored information. Several members of the Src family of tyrosine kinases are activated by stimulation of the NMDAr and are involved in both induction of hippocampal long-term potentiation and consolidation of hippocampal-dependent, NMDAr-sensitive, memories. Here we analyzed the role of the Src family within the CA1 region of the hippocampus in extinction and reacquisition of the memory for step-down, inhibitory avoidance learning task (IA). Rats trained in IA were submitted to 5 daily extinction sessions during which the avoidance response was elicited in the absence of the unconditioned stimulus. Immediately or 180 min after each extinction session animals received intra-CA1 infusions of either 0.1% DMSO, the Src-family inhibitor PP2 or its inactive analog, PP3. PP2 blocked extinction of the IA response which was otherwise evident in DMSO and PP3-treated animals. After being submitted to a new training session the animals reacquired the avoidance response; however, they failed to do so if they received intra-CA1 infusions of PP2 immediately following retraining. Our results indicate that, like the original learning, extinction and reacquisition of the IA response require activity of the Src family in the CA1 region of the hippocampus. PMID- 15894072 TI - Electroacupuncture combined with MK-801 prolongs anti-hyperalgesia in rats with peripheral inflammation. AB - Our previous study showed that electroacupuncture (EA), an adjuvant to conventional medicine, significantly attenuated hyperalgesia in a rat model of inflammatory pain. In the present study, we evaluated the potential additive and/or synergism of EA and a sub-effective dose of dizocilpine maleate (MK-801), a non-competitive N-methyl-d-aspartate (NMDA) receptor antagonist, on hyperalgesia in the same rat model of inflammatory pain. Hyperalgesia, manifesting as decreased paw withdrawal latency (PWL) to a noxious stimulus, was induced by injecting complete Freund's adjuvant (CFA) into the plantar surface of one hind paw of each rat. EA treatments were given at acupoint GB30 immediately after and 2 h after CFA. MK-801 at 0.001 mg/rat was given (i.t.) 10 min before each of the two EA treatments. PWL was measured prior to and 2.5 and 5 h post CFA. Ten and 100 Hz EA significantly inhibited CFA-induced hind paw hyperalgesia. Both 10 and 100 Hz EA combined with the sub-effective dose of 0.001 mg/rat MK-801 showed prolonged anti-hyperalgesia with no side effects. These results demonstrate that EA combined with a sub-effective dose of this NMDA receptor antagonist enhances anti-hyperalgesia, and this combination may provide an effective strategy for pain management. PMID- 15894073 TI - Involvement of hypothalamic pituitary adrenal axis on the effects of nifedipine in the development of morphine tolerance in rats. AB - It has been shown that nifedipine, as a calcium channel blocker, can attenuate the development of tolerance to the antinociceptive effect of morphine; however, the role of HPA axis on this action has not been elucidated. We examined the effect of nifedipine on morphine analgesic tolerance in intact and adrenalectomized (ADX) rats and on HPA activity induced by morphine. Adult male rats were rendered tolerant to morphine by daily injection of morphine (15 mg/kg i.p.) for 8 days. To determine the effect of nifedipine on the development of morphine tolerance, nifedipine (1, 2 and 5 mg/kg i.p.) was injected concomitant with morphine. The tail-flick test was used to assess the nociceptive threshold, before and 30 min after morphine administration in days 1, 3, 5 and 8. Our results showed that despite the demonstration of tolerance in both ADX and sham operated rats, nifedipine in ADX rats prevented morphine tolerance development at a lower dose (2 mg/kg) than in sham operated rats, however corticosterone replacement prevented nifedipine effect in ADX rats. Acute administration of morphine produced significant increase in plasma corticosterone level, and with repeated injection, a tolerance to this neurosecretory effect was developed. Nifedipine (5 mg/kg) attenuated the acute effect of morphine, but could not block its neurosecretory tolerance. PMID- 15894074 TI - Placebo cigarettes in a spaced smoking paradigm. AB - Existing evidence supports the notion that nicotine delivery and recentness of smoking mediate the effects of smoking, including decreases in tobacco craving. However, smoking placebo (denicotinized) cigarettes decreases tobacco craving after overnight abstinence. The present study tested whether the recentness of smoking was an important determinant in the ability of a placebo cigarette to reduce tobacco craving. Placebo (0.07 mg nicotine) and conventional (1.1 mg nicotine) cigarettes were used in a spaced smoking paradigm. In six experimental sessions lasting 240 min, subjects smoked either a placebo or conventional nicotine cigarette in intervals of either 30, 60, or 240 min. Heart rate (HR), exhaled carbon monoxide (CO) levels, and subjective (Schuh-Stitzer, QSU) measures of tobacco craving were obtained throughout the spaced smoking paradigm. HR and CO levels increased after smoking both types of cigarettes. Increasing the interval since the last cigarette significantly (p<0.001) increased the baseline values of tobacco craving. Smoking either the placebo or the conventional cigarette caused a significant (p<0.01) reduction in the craving score after smoking. However, the nicotine yield of the cigarette did not influence these patterns. It is concluded that acute tobacco cravings can be repeatedly diminished with cigarettes that do not deliver nicotine. PMID- 15894075 TI - Mecamylamine attenuates ephedrine-induced hyperactivity in rats. AB - Ephedrine is a central nervous system stimulant that has a pharmacological profile similar to amphetamines. Ephedrine induces hyperactivity after acute administration to rats and locomotor sensitization develops to ephedrine with repeated administration. Recent research suggests that nicotinic receptors (nAChRs) play a role in the development of locomotor sensitization to d amphetamine and the goal of the present study was to determine if nAChRs similarly mediate the effects of ephedrine after acute and repeated drug injection. On 12 consecutive days, rats were pretreated with the nAChR antagonist mecamylamine (0.3-3.0 mg/kg) or saline followed by (-)-ephedrine (10-30 mg/kg) or saline injection and locomotor activity was measured. Ephedrine produced a dose dependent increase in locomotor activity, and sensitization to ephedrine developed with repeated injection. Mecamylamine pretreatment attenuated the hyperactivity and sensitization produced by repeated, but not acute, ephedrine (10 mg/kg) injection. The inhibitory effect of mecamylamine was overcome at the higher ephedrine dose (30 mg/kg). The present results indicate that nAChRs play a mediating role in the development of locomotor sensitization to ephedrine. PMID- 15894076 TI - Behavioral and pharmacological characterization of a distal peripheral nerve injury in the rat. AB - Previous rat neuropathic pain models have utilized peripheral nerve injuries that damage a significant proportion of large nerves such as the sciatic nerve or its divisions. Injuries that lead to neuropathic pain in humans may involve the peripheral extremities. The current study evaluated the behavioral effects of injury to the plantar nerves in the rat (distal nerve injury-DNI). A delayed onset of hypersensitivity to an innocuous mechanical stimulus was observed following cutting of the left plantar nerves, whereas mechanical hypersensitivity developed more rapidly in rats with either an injury near the sciatic nerve trunk (chronic constriction injury (CCI), spared nerve injury (SNI)) or a spinal nerve root (spinal nerve ligation (SNL). Similar to other nerve injury pain models, rats with injured plantar nerves also developed an early onset and persistent sensitivity to a cooling stimulus. The effects of morphine, gabapentin and imipramine on mechanical and cold hypersensitivity were evaluated in rats with a DNI, CCI and SNI. In all three models, morphine dose-dependently suppressed mechanical and cold hypersensitivity, whereas gabapentin only suppressed mechanical hypersensitivity. Imipramine had no effect on either cold or mechanical hypersensitivity in any of the nerve-injured rats. The pharmacological data suggest that the underlying basis of neuropathic pain may be similar irrespective of the site of nerve injury. PMID- 15894077 TI - Recovery from ischemic brain injury in the rat following a 10 h delayed injection with MLN519. AB - In the present study, we evaluated delayed treatment effects of the proteasome inhibitor and anti-inflammatory agent MLN519 (initiated 10 h post-injury) to improve recovery following ischemic brain injury in rodents. Male rats were exposed to 2 h of middle cerebral artery occlusion (MCAo) and treated with MLN519 (1.0 mg/kg, i.v. @ 10, 24, and 48 h post-occlusion) or vehicle. By 2 weeks post injury, 60% (6/10) of vehicle animals survived, which was improved (although non significantly) to 78% (7/9) following MLN519 treatment. The percent loss of tissue in the ipsilateral brain hemisphere (at 2 weeks) was significantly reduced from 27+/-4% (vehicle) to 15+/-4% (MLN519). MLN519 treated animals also lost significantly less body weight (39%) and showed significant improvement in overall neurological function across the 2-week recovery period. However, no significant treatment effects were observed to reduce foot-fault deficits (balance beam) or improve recovery of operant performance (active avoidance test). Overall, delayed treatment with MLN519 provided significant improvement in 3 of 6 test metrics (histopathology, body weight, and neurological dysfunction) supporting improved outcome for brain-injured subjects. PMID- 15894078 TI - The selective dopamine D3 receptor antagonist SB-277011-A attenuates ethanol consumption in ethanol preferring (P) and non-preferring (NP) rats. AB - The mesolimbic dopamine (DA) system plays an important role in mediating addiction to alcohol and other drugs of abuse. Recent evidence points toward the role of the DA D3 receptor (D3R) in drug-induced reward, drug-taking, as well as cue-, drug-, and stress-triggered relapse to drug-seeking behavior. Accordingly, the present study examined the effects of acute selective antagonism of the D3R on ethanol consumption in alcohol Preferring (P) and Non-Preferring (NP) rats. We employed the two-bottle choice paradigm to monitor ethanol consumption in these rats before and after treatment with 3, 10, and 30 mg/kg (i.p.) of the selective D3R antagonist SB-277011-A. Results indicated a significant attenuation in ethanol preference, intake and lick responses in P rats treated with 10 and 30 mg/kg SB-277011-A. A similar, though not as robust effect was observed in ethanol consumption in the NP rats when treated with 30 mg/kg SB-277011-A. Finally, the acute administration of SB-277011-A did not produce extrapyramidal side effects, as indicated by stable lick response-volume ratios and lick response time distributions. These results further support the notion that the D3R is important in mediating the addictive properties of alcohol and suggest that selective blockade of the D3R may constitute a new and useful target for prospective pharmacotherapeutic approaches to alcoholism. PMID- 15894079 TI - Relation between methamphetamine-induced monoamine depletions in the striatum and sequential motor learning. AB - Methamphetamine (METH) use results in depletion of monoamines in the striatum. The purpose of this study was to evaluate the relation between the degree of METH induced monoamine depletion in the striatum and impairment on a striatally dependent learning task in rats. Male Sprague-Dawley rats received four injections of METH (10 mg/kg) or saline at 2-h intervals. METH treatment produced a 38.5% (+/-5.6) and 46.7% (+/-6.7) dopamine (DA) depletion in the medial and lateral striatum, respectively. Serotonin (5-HT) was depleted 15.6% (+/-10.4) and 21.1% (+/-8.2) in the medial and lateral striatum, respectively. One month after treatment, rats were trained on a sequential-memory task on an 8-arm radial maze. METH-treated rats made significantly fewer direct movements between arms in the maze sequence across days of trials. The learning impairment was significantly correlated with the degree of DA depletion in the medial striatum, as well as serotonin tissue content in striatum. Only rats with a greater than 40% DA depletion in medial striatum showed significant impairments. These results provide additional evidence for METH-induced learning impairments and suggest that this impairment is dependent on the striatal monoamine loss, in general, and the degree of DA loss in medial striatum, in particular. PMID- 15894080 TI - Effect of chronic Albizzia julibrissin treatment on 5-hydroxytryptamine1A receptors in rat brain. AB - Quantitative receptor autoradiography and behavioral studies were employed to investigate whether the aqueous extract of Albizzia julibrissin (AEAJ) specifically targets serotonergic systems in rat brain. AEAJ was orally administered at 50 and 200 mg/kg to adult male SD rats for 7 days. Treatment with AEAJ (200 mg/kg) significantly increased time-spent in open arms and the number of open arm entries in an elevated plus-maze (EPM) versus saline controls (P<0.05). Moreover, those effects of AEAJ were blocked by WAY 100635, a 5-HT1A receptor antagonist. Following behavioral evaluation, the binding of [3H]8-hyroxy 2-(di-n-propylamino) tertalin ([3H]8-OH-DPAT) to 5-HT1A receptors in rat brain was investigated. [3H]8-OH-DPAT binding after AEAJ (200 mg/kg) treatment showed a marked increase in the frontal cortex, hippocampus (CA2 and CA3 regions) and in the lateral septum versus vehicle-treated controls. No changes of [3H]8-OH-DPAT binding were observed in the caudate putamen, dentate gyrus and CA1 areas of the hippocampus or in the hypothalamus. In the dorsal raphe region, [3H]8-OH-DPAT binding was significantly reduced by AEAJ (50 mg/kg) treatment but was unchanged by AEAJ (200 mg/kg). These results suggest that the anxiolytic-like effect of A. julibrissin is mediated by the changes of serotonergic nervous system, especially 5-HT1A receptors. PMID- 15894081 TI - Dopamine D2, but not D4, receptor agonists are emetogenic in ferrets. AB - Agents that activate the dopamine D2-like family of receptors elicit emesis in humans and other species with a vomiting/emetic reflex; however, the lack of dopamine receptor subtype selective agonists has hampered an understanding of which dopamine D2-like receptor subtype(s) contributes to the emetic response. In this study, stable cell lines expressing the ferret dopamine D2-long (D2L) and D4 receptors were used to characterize known dopamine agonists via radioligand binding and calcium ion flux assays, while emetic activity of these dopamine receptor agonists was determined in male ferrets. Latencies to first emetic event, average number of emetic episodes, and stereotypical behaviors which may be indicative of nausea were also determined. Agonists at dopamine D1-like and D4 receptors had no emetic effect in ferrets. Conversely, stimulation of dopamine D2 and/or D3 receptors resulted in a robust emetic response characterized by a relatively short latency (<15 min) and multiple emetic events. Competitive antagonists of dopamine D2-like receptors (domperidone, haloperidol) dose dependently blocked the emetic response to PNU95666E, a dopamine D2 receptor selective agonist. Thus, dopamine D2 and/or D3 receptor agonists elicit emesis, while dopamine D1/D5 or D4 receptor-selective agonists are devoid of emetic properties. PMID- 15894082 TI - The 2000 EBCTCG overview: a widening gap. PMID- 15894083 TI - Laparoscopic-assisted resection of colorectal carcinoma. PMID- 15894084 TI - Enhanced access to emergency contraception. PMID- 15894085 TI - Missed contraceptive pills and the critical pill-free interval. PMID- 15894086 TI - Tailoring breast cancer therapy to genetic status. PMID- 15894087 TI - The National Service Framework: paediatric emergency care. PMID- 15894090 TI - Prediction of cancer outcome with microarrays. PMID- 15894091 TI - Prediction of cancer outcome with microarrays. PMID- 15894093 TI - Prediction of cancer outcome with microarrays. PMID- 15894094 TI - Prediction of cancer outcome with microarrays. PMID- 15894095 TI - Prediction of cancer outcome with microarrays. PMID- 15894097 TI - Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. AB - BACKGROUND: Quinquennial overviews (1985-2000) of the randomised trials in early breast cancer have assessed the 5 year and 10-year effects of various systemic adjuvant therapies on breast cancer recurrence and survival. Here, we report the 10-year and 15-year effects. METHODS: Collaborative meta-analyses were undertaken of 194 unconfounded randomised trials of adjuvant chemotherapy or hormonal therapy that began by 1995. Many trials involved CMF (cyclophosphamide, methotrexate, fluorouracil), anthracycline-based combinations such as FAC (fluorouracil, doxorubicin, cyclophosphamide) or FEC (fluorouracil, epirubicin, cyclophosphamide), tamoxifen, or ovarian suppression: none involved taxanes, trastuzumab, raloxifene, or modern aromatase inhibitors. FINDINGS: Allocation to about 6 months of anthracycline-based polychemotherapy (eg, with FAC or FEC) reduces the annual breast cancer death rate by about 38% (SE 5) for women younger than 50 years of age when diagnosed and by about 20% (SE 4) for those of age 50 69 years when diagnosed, largely irrespective of the use of tamoxifen and of oestrogen receptor (ER) status, nodal status, or other tumour characteristics. Such regimens are significantly (2p=0.0001 for recurrence, 2p<0.00001 for breast cancer mortality) more effective than CMF chemotherapy. Few women of age 70 years or older entered these chemotherapy trials. For ER-positive disease only, allocation to about 5 years of adjuvant tamoxifen reduces the annual breast cancer death rate by 31% (SE 3), largely irrespective of the use of chemotherapy and of age (<50, 50-69, > or =70 years), progesterone receptor status, or other tumour characteristics. 5 years is significantly (2p<0.00001 for recurrence, 2p=0.01 for breast cancer mortality) more effective than just 1-2 years of tamoxifen. For ER-positive tumours, the annual breast cancer mortality rates are similar during years 0-4 and 5-14, as are the proportional reductions in them by 5 years of tamoxifen, so the cumulative reduction in mortality is more than twice as big at 15 years as at 5 years after diagnosis. These results combine six meta analyses: anthracycline-based versus no chemotherapy (8000 women); CMF-based versus no chemotherapy (14,000); anthracycline-based versus CMF-based chemotherapy (14,000); about 5 years of tamoxifen versus none (15,000); about 1-2 years of tamoxifen versus none (33,000); and about 5 years versus 1-2 years of tamoxifen (18,000). Finally, allocation to ovarian ablation or suppression (8000 women) also significantly reduces breast cancer mortality, but appears to do so only in the absence of other systemic treatments. For middle-aged women with ER positive disease (the commonest type of breast cancer), the breast cancer mortality rate throughout the next 15 years would be approximately halved by 6 months of anthracycline-based chemotherapy (with a combination such as FAC or FEC) followed by 5 years of adjuvant tamoxifen. For, if mortality reductions of 38% (age <50 years) and 20% (age 50-69 years) from such chemotherapy were followed by a further reduction of 31% from tamoxifen in the risks that remain, the final mortality reductions would be 57% and 45%, respectively (and, the trial results could well have been somewhat stronger if there had been full compliance with the allocated treatments). Overall survival would be comparably improved, since these treatments have relatively small effects on mortality from the aggregate of all other causes. INTERPRETATION: Some of the widely practicable adjuvant drug treatments that were being tested in the 1980s, which substantially reduced 5-year recurrence rates (but had somewhat less effect on 5-year mortality rates), also substantially reduce 15-year mortality rates. Further improvements in long-term survival could well be available from newer drugs, or better use of older drugs. PMID- 15894098 TI - Short-term endpoints of conventional versus laparoscopic-assisted surgery in patients with colorectal cancer (MRC CLASICC trial): multicentre, randomised controlled trial. AB - BACKGROUND: Laparoscopic-assisted surgery for colorectal cancer has been widely adopted without data from large-scale randomised trials to support its use. We compared short-term endpoints of conventional versus laparoscopic-assisted surgery in patients with colorectal cancer to predict long-term outcomes. METHODS: Between July, 1996, and July, 2002, we undertook a multicentre, randomised clinical trial in 794 patients with colorectal cancer from 27 UK centres. Patients were allocated to receive laparoscopic-assisted (n=526) or open surgery (n=268). Primary short-term endpoints were positivity rates of circumferential and longitudinal resection margins, proportion of Dukes' C2 tumours, and in-hospital mortality. Analysis was by intention to treat. This trial has been assigned the International Standard Randomised Controlled Trial Number ISRCTN74883561. FINDINGS: Six patients (two [open], four [laparoscopic]) had no surgery, and 23 had missing surgical data (nine, 14). 253 and 484 patients actually received open and laparoscopic-assisted treatment, respectively. 143 (29%) patients underwent conversion from laparoscopic to open surgery. Proportion of Dukes' C2 tumours did not differ between treatments (18 [7%] patients, open vs 34 [6%], laparoscopic; difference -0.3%, 95% CI -3.9 to 3.4%, p=0.89), and neither did in-hospital mortality (13 [5%] vs 21 [4%]; -0.9%, -3.9 to 2.2%, p=0.57). Apart from patients undergoing laparoscopic anterior resection for rectal cancer, rates of positive resection margins were similar between treatment groups. Patients with converted treatment had raised complication rates. INTERPRETATION: Laparoscopic-assisted surgery for cancer of the colon is as effective as open surgery in the short term and is likely to produce similar long term outcomes. However, impaired short-term outcomes after laparoscopic-assisted anterior resection for cancer of the rectum do not yet justify its routine use. PMID- 15894099 TI - Breast cancer. AB - Breast cancer remains a public-health issue on a global scale. We report new information about the disease from the past 5 years. Early age at first birth, increasing parity, and tamoxifen use are related to long-term lifetime reduction in breast-cancer risk. Ductal carcinomas in situ has been suggested to be renamed ductal intraepithelial neoplasia to emphasise its non-life-threatening nature. An alternative approach, the progenitor/stem cell theory, predicts that only some tumour cells cause cancer progression and that these should be targeted by treatment. Mammography and ultrasonography are still the most effective for women with non-dense and dense breast tissues, respectively. Additionally, MRI, lymphatic mapping, the nipple-sparing mastectomy, partial breast irradiation, neoadjuvant systemic therapy, and adjuvant treatments are promising for subgroups of breast-cancer patients. Although tamoxifen can be offered for endocrine responsive disease, aromatase inhibitors are increasingly used. Assessment of potential molecular targets is now important in primary diagnosis. Tyrosine kinase inhibitors and other drugs with anti-angiogenesis properties are currently undergoing preclinical investigations. PMID- 15894100 TI - Breast cancer. PMID- 15894101 TI - An act of unkindness. PMID- 15894102 TI - A girl with persistent hyperferritinaemia. PMID- 15894103 TI - Immunology: turning basic research into a therapy. AB - One of the driving forces in the field of immunology is the ambition to translate experimental research into novel useful therapies. Therefore, the aim of this mini-review is to exemplify emerging therapies as well as highlight hurdles that need to be overcome before they can be introduced into the clinic. PMID- 15894104 TI - Dendritic cells in tolerance induction. AB - Regulatory T cells (Treg) in broader terms consist of different subsets of T cells that are characterized by their ability to suppress proliferation of conventional effector T cells by various means. To date three main groups of Treg can de distinguished, mainly by their functional properties. The regulatory T cell family: distinct subsets and their interrelations. Briefly, T regulatory (Tr)-1 cells as well as T helper (Th)-3 T cells express common T cell markers such as CD4 and are characterized by secretion of IL-10 and TGF-beta, which provides a means by which proliferation of conventional CD4(+) cells is blocked. In contrast, genuine Treg that are characterized by their expression of CD25 block T cell proliferation by an unknown cell to cell contact dependent mechanism. However, there are many overlapping features shared by the different subtypes of regulatory T cells and the common denominator is the production of regulatory cytokines such as IL-10 and TGF-beta. PMID- 15894105 TI - Who controls the crowd? New findings and old questions about the intestinal microflora. AB - It is probably one of the most complex tasks to understand the mutual relationship between members of the microbial community in the gut and their interaction with the host. Although many animals including humans can live without gut flora, it is beneficial for the growth and protection of the host against pathogens. On the other hand, clinical and experimental intestinal inflammatory reactions have been attributed to immune recognition of the intestinal microflora. This review discusses recent and old findings on function and immunological relevance of the endogenous microflora. PMID- 15894106 TI - T helper (h)1/Th2 and Leishmania: paradox rather than paradigm. AB - Studies on Leishmania major have been largely responsible for the characterisation of the Th1/Th2 paradigm of healing/non-healing associated with intracellular infection. IFN-gamma and IL-4 were identified respectively as the counter-regulatory Th1 and Th2 cytokines promoting resistance and susceptibility to infection. While resistance against infection remains largely associated with an IL-12 induced type-1 response studies using in particular gene-deficient mice have questioned the paramount role of IL-4 in the non-healing disease and implicated several alternative candidates. Indeed IL-4 has been shown to have no exacerbatory role in murine visceral leishmaniasis while its contribution to the progression of cutaneous disease has been clearly shown to be influenced not only by the parasite species but also the mouse strain used. Furthermore, it is now well established that not only can Th2 responses be induced independently of IL-4 but IL-4 under certain circumstances can prime for IL-12 production and a type-1 response. Clearly a reappraisal of our current understanding of the role of IL-4 and the Th2 response in the immunobiology of leishmaniasis is required and this review seeks to address this issue. PMID- 15894107 TI - Inhibition of interleukin-12 production in mouse macrophages by hydroquinone, a reactive metabolite of benzene, via suppression of nuclear factor-kappaB binding activity. AB - Exposure to cigarette smoke is known to increase the risk of the development of allergic disease associated with T helper type 2 (Th2)-mediated immune responses. IL-12 is known to suppress Th2 responses. In this study we investigated the effects of hydroquinone (HQ), a major metabolite of benzene present in large quantities in cigarette tar, on the production of IL-12 from mouse macrophages stimulated with lipopolysaccharide (LPS). HQ potently inhibited the LPS-induced IL-12 production in both primary mouse macrophages and RAW164.7 monocytic cells in a dose-dependent manner. The effect of HQ on IL-12 p40 promoter activation was analyzed by transfecting RAW264.7 monocytic cells with p40 promoter/luciferase constructs. The repressive effect mapped to a region in the p40 promoter containing a binding site for nuclear factor-kappaB (p40-kappaB). Furthermore, activation of macrophages by LPS resulted in markedly enhanced binding activity to the kappaB site, which significantly decreased upon addition of HQ. Pre incubation with HQ significantly prevented degradation of IkappaB protein in LPS stimulated macrophage cells, indicating that HQ suppressed NF-kappaB binding activity by inhibiting the degradation of IkappaB protein. These findings suggest that HQ may, at least in part, enhance allergic immune responses by inhibiting the production of IL-12 in macrophages. PMID- 15894108 TI - Interleukin 18 (IL-18) upregulation in acute rejection of kidney allograft. AB - Interleukin 18 (IL-18) is a potent proinflammatory cytokine involved in the host defence by upregulating both innate and acquired immune responses and may be of particular importance also in mechanisms of kidney allograft rejection. Immunohistochemical staining of protocol biopsies showed constitutive IL-18 expression in the epithelium of distal tubules with the induction of immunoreactivity in acute rejection patients where also proximal tubules, infiltrating leukocytes, and endothelium were strongly positive. Furthermore, serum levels of IL-18 were significantly elevated in patients with acute rejection of kidney allograft (1247+/-389 pg/l) as compared to patients with uncomplicated outcome of kidney transplantation (444+/-164 pg/l) and subjects with acute tubulointerstitial nephropathy (385+/-155 pg/l, p<0.0001 for both comparisons). Tissue culture model of renal epithelial cells expressed IL-18 mRNA constitutively and released mature IL-18 in response to TNF-alpha and IFN-gamma. We assume that upregulation of epithelial IL-18 plays an important role in immune and immunopathological reactions in renal parenchyma and contributes to rejection mechanisms of kidney allograft. PMID- 15894109 TI - Intestinal TSH production is localized in crypt enterocytes and in villus 'hotblocks' and is coupled to IL-7 production: evidence for involvement of TSH during acute enteric virus infection. AB - The immune and neuroendocrine systems have been shown to work conjointly in a number of ways. One aspect of this has to do with a potential role for thyroid stimulating hormone (TSH) in the regulation of the mucosal immune system, although the mechanisms by which this occurs remain vague. To more thoroughly understand how TSH participates in intestinal intraepithelial lymphocyte (IEL) development and immunity, experiments have been conducted to define local sites of intestinal TSH production, and to characterize changes that occur in the synthesis of TSH during acute enteric virus infection. Here, we demonstrate that TSH in the small intestine is specifically localized to regions below villus crypts as seen by immunocytochemical staining, which revealed high-level TSH staining in lower crypts in the absence of IL-7 staining, and TSH and IL-7 co staining in upper crypt regions. Additionally, prominent TSH staining was evident in TSH 'hotblocks' sparsely dispersed throughout the epithelial layer. In rotavirus-infected mice, the TSH staining pattern differed significantly from that of non-infected animals. Notably, at 2 and 3 days post-infection, TSH expression was high in and near apical villi where virus infection was greatest. These findings lend credence to the notion that TSH plays a role both in the development of intestinal T cells, and in the process of local immunity during enteric virus infection. PMID- 15894110 TI - LY294002 inhibits LPS-induced NO production through a inhibition of NF-kappaB activation: independent mechanism of phosphatidylinositol 3-kinase. AB - Phosphatidylinositol 3-kinase (PI3K) is critical player in cell proliferation and survival. The effects of LY294002 and wortmannin, inhibitors of PI3K, on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipoploysaccharide (LPS)-induced Raw 264.7 cells were investigated. Significant inhibition of LPS-induced protein kinase B (PKB, Akt) phosphorylation occurred at 25 microM LY294002 or 0.5 microM wortmannin. At the same concentrations, LY294002, but not wortmannin, significantly inhibited NO production and iNOS expression. LY303511, an inactive analogue of LY294002, also inhibited NO production and iNOS expression. In addition, LY294002 and LY303511 significantly inhibited the DNA binding activity of NF-kappaB and NF-kappaB dependent reporter gene expression. These results suggest that LY294002 inhibits iNOS expression at least in part via inhibition of NF-kappaB activation, independent of PI3K. PMID- 15894111 TI - Dramatic changes in the serum levels of anti-cholesterol antibodies after eversion endarterectomy in patients with severe carotid atherosclerosis. AB - Our goal was to study changes in anti-cholesterol antibodies (ACHA) levels in patients with severe carotid stenosis after eversion endarterectomy. Seventy consecutive patients who underwent eversion endarterectomy at the Department of Cardiovascular Surgery, Semmelweis University, Budapest, were included in the study. Serum samples from 66 healthy volunteers served as controls. Patients had medical check up at 5.7 (4.6-8.0) weeks (median (interquartile range)), 6.8 (6.2 7.9) months as well as 13.8 (12.3-19.0) months after endarterectomy. In all patients the carotid arteries were investigated by color duplex ultrasound. ACHA concentrations were determined in the serum samples taken before operation as well as at the first and last follow-up visits by using an ELISA method. ACHA concentrations (median (interquartile range) were found to be significantly (p<0.0001) lower in the sera of the patients with carotid atherosclerosis (13.5 (8.4-21.3)AU/ml) than in the healthy subjects (26.1 (20.9-33.2)AU/ml) (Mann Whitney test). Strong negative correlation was found between the preoperative ACHA and LDL-cholesterol levels (r=-0.413, p=0.0004). Serum ACHA concentrations significantly (p<0.0001) increased from the values measured before operation (13.5 (8.4-21.3)AU/ml) to 27.1 (19.9-34.7)AU/ml measured at the end of the 14 months long follow-up. Increase occurred only in patients with low or medium baseline ACHA concentration. Our present findings indicate that after surgical removal of atherosclerotic plaques from the carotid arteries the reduced baseline levels of ACHA reach normal values in 1 year. Different not mutually exclusive mechanisms (binding of ACHA to lipid/lipoproteins particles, to advanced plaques or reversibly injured endothelial cells in CNS) can be responsible for this novel finding. PMID- 15894112 TI - Predominance of Type 1 cytokines and decreased number of CD4(+)CD25(+high) T regulatory cells in peripheral blood of patients with recurrent aphthous ulcerations. AB - Recurrent aphthous ulcerations (RAU) are a chronic inflammatory disease with evidence of inappropriate immune response. Previous studies have suggested cell mediated activation of immune response towards common micro-organisms of oral cavity in RAU. In this investigation, we explored cytokine production by peripheral blood mononuclear cells (PBMC) and T regulatory cell population in blood of active and remission RAU patients as crucial factors for maintenance of peripheral tolerance. Ten patients with minor RAU and 12 healthy individuals were selected for the study. Cytokine levels were analysed in supernatants using Cytometric Bead Array Kit for flow cytometry and ELISA. We have demonstrated increased production of Type 1 cytokines IL-2, IFN-gamma and TNF-alpha as well as IL-5, IL-6 and IL-8 by peripheral blood mononuclear cells in RAU. In contrast, IL 10 and TGF-beta anti-inflammatory cytokine production was decreased in RAU patients compared to healthy individuals. Moreover, we have found that CD4(+)CD25(+high) T regulatory cell proportion was decreased in RAU and represented 3.58+/-0.654% of CD4(+) T cells in active RAU, 4.66+/-0.561% of CD4(+) T cells in remission RAU, whereas in healthy controls CD4(+)CD25(+high) T cells represented 7.30+/-1.238% of CD4(+) T cells (p<0.001). Thus, the obtained results indicate that disproportion in cytokine production may be contributing factor in the pathogenesis of RAU. Alteration in the number of CD4(+)CD25(+high) T regulatory cells in RAU may additionally influence the development of the disease. We propose that imbalance in pro- and anti-inflammatory cytokine network may lead to the breakdown of peripheral tolerance in RAU and the excessive immune response towards harmless micro-organisms colonized oral mucosa or self-antigens. PMID- 15894113 TI - Identification of NH(2)-terminal amino acid residues essential for the biological activity of leukotactin-1. AB - Leukotactin-1 (Lkn-1), a human CC chemokine that binds to both CC chemokine receptor (CCR)1 and CCR3, is distinct from other human CC chemokines in that it has long amino acid residues preceding the first cysteine at the NH(2)-terminus. Serial deletion studies showed that at least three amino acid residues, alanine alanine-aspartic acid (A-A-D), preceding the first cysteine at the NH(2)-terminus are essential for the biological activity of Lkn-1. Point mutation and deletion studies for the three amino acids were performed in the present study. Substitutions of the first alanine residue with other amino acids did not cause significant loss of biological activities. Deletion of the third amino acid, aspartic acid, resulted in more than 100-fold loss of the activity. Deletion of two amino acids, alanine-alanine (A-A) or alanine-aspartic acid (A-D), resulted in almost complete loss of the activity. Loss of agonistic activity by deletion of two amino acids was due to impaired binding to CCR1. These results identify that alanine-aspartic acid residues preceding the first cysteine at the NH(2) terminus are essential for the binding and biological activity of Lkn-1. PMID- 15894114 TI - Localization in membrane microdomains of the Ig-alpha/beta component of the BCR expressed by a B lymphoma variant. AB - The B cell antigen receptor (BCR) composed of the ligand-binding membrane IgM (mIgM) and the signaling component, Ig-alpha/beta, is known to inducibly associate with membrane microdomains rich in cholesterol and sphingolipids, termed lipid rafts. In this study we tested whether the Ig-alpha/beta portion of the BCR has targeting information that allows it to be localized in lipid rafts. In order to do this, we cross-linked the Ig-alpha/beta on the cell surface of the variant B cell line, WEHI 303.1.5, a derivative of the immature murine B cell line WEHI 231 that lacks mu heavy chain and expresses the Ig-alpha/beta on the cell surface by itself. Using two methods to isolate detergent-insoluble, lipid raft-like fractions, we found that Ig-alpha/beta without accompanying mIgM was constitutively located in these raft-like fractions and that the amount was marginally increased after Ig-alpha/beta cross-linking. These results suggest that the Ig-alpha/beta portion of the BCR has the ability to be compartmentalized into raft-like membrane domains even when not associated with mIgM and perhaps this targeting information is normally regulated by the presence of the mIgM portion of the receptor. PMID- 15894115 TI - PKC- and PI3K-dependent but ERK-independent proliferation of murine splenic B cells stimulated by chondroitin sulfate B. AB - High molecular weight polyanions such as dextran sulfate are known to be weak polyclonal activators of murine B cells, but the molecular mechanism of their mitogenic activitiy is not fully elucidated. Although chondroitin sulfate A (CSA), B (CSB) and C (CSC) are highly charged polyanions, little is known about their effects on the proliferation of B cells. In this study, we demonstrated that CSB stimulated proliferation of murine B cells as markedly as did anti-IgM antibody, more markedly than did dextran sulfate and much more markedly than did CSA, CSC, heparin and hyaluronic acid. CSB caused translocation of protein kinase C (PKC) isoform beta from cytosol to membrane fractions and increased phosphorylation of Akt but not phosphorylation of extracellular signal-regulated kinase (ERK) of B cells. CSB-induced B cell proliferation was almost completely blocked by either the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 or the PKC inhibitor GF109203X but was not significantly inhibited by the ERK kinase inhibitor PD98059. The mitogenic effect of anti-IgM was significantly inhibited by all the three inhibitors, while the mitogenic effect of LPS was inhibited only by LY294002. These findings indicate that CSB stimulated proliferation of murine B cells more markedly than did dextran sulfate and suggest that PKC and PI3K are crucial but that ERK is less important for the mitogenic activity of CSB, the signaling pathways of which may be at least partly distinct from those of anti IgM and LPS. PMID- 15894116 TI - Specific antitumor immune response induced by a novel DNA vaccine composed of multiple CTL and T helper cell epitopes of prostate cancer associated antigens. AB - The loss of immunogenic epitopes by tumors has urged the development of vaccines against multiple epitopes. Recombinant DNA technologies have opened the possibility to develop multiepitope vaccines in a relatively rapid and efficient way. In this study, several DNA fragments encoding multiple cytotoxic T lymphocyte (CTL) and T helper (Th) cell epitopes were selected from human prostate-specific membrane antigen (hPSM), mouse prostatic acid phosphatase (mPAP), and human prostate-specific antigen (hPSA), These DNA fragments were ligated together to form a novel fusion gene, termed 3P gene. The 3P gene and human IgG Fc gene were inserted into pcDNA3.1 to construct a DNA vaccine designated psig-3P-Fc. Vaccination with psig-3P-Fc by gene gun inoculation induced strong antitumor response in a mouse tumor model, which significantly inhibited tumor growth and prolonged survival time of the tumor-bearing mice. In vitro, when lymphocytes were stimulated by psig-3P-Fc-transfected autologous peripheral blood mononuclear cells (PBMC), CTLs were induced which could specifically kill hPSM-, hPAP-, or hPSA-expressing tumor cells. These observations provide a new vaccine strategy for cancer therapy through concomitant enhancement of antigen specific CD4(+) helper and CD8(+) cytotoxic T cell responses against tumors. PMID- 15894117 TI - Trephine biopsies are enriched for activated T/NK cells and cytotoxic T cells. AB - Although bone marrow aspiration is the most commonly obtained human marrow sample type, it yields liquid samples that contain a varying degree of blood contamination. Trephine biopsies, on the other hand, are solid marrow cores and are, therefore, much less likely to contain contaminating peripheral blood. In this study, we utilised a technique to extract viable cells from solid trephine biopsy specimens, by means of mechanical and enzymatic digestion, allowing cytometric comparison of cells in these biopsies and simultaneously obtained liquid aspirate samples. Having established that the digestion procedure itself was not causing any significant alteration in the immunophenotypic composition of the marrow samples, our data show that trephine biopsies were enriched for CD8(+) T cells, with concomitant decrease in the CD4(+) subset, compared to paired aspirates. Furthermore, T cells, NK cells and T cells expressing NK cell receptor (NKR) molecules were all significantly more likely to express both early (CD69) and late (HLA-DR) markers of activation. Bone marrow aspirates do not, therefore, provide truly representative data on the phenotypic composition of bone marrow, and the effect of peripheral blood contamination in aspirates should be taken into account when comparisons are being made between the bone marrow and other human issues or, perhaps more so, between human and murine marrow. PMID- 15894118 TI - Coculture with dendritic cells promotes proliferation but not cytotoxic activity of gamma/delta T cells. AB - T cells bearing the gamma9/delta2 T cell receptor (TCR) have recently raised interest as non-MHC restricted effector cells against multiple myeloma. They are described to be stimulated by phosphoantigens without the need of antigen presenting cells. However, in the past a positive effect of cells of the monocyte lineage on activation of gamma/delta T cells has been shown. Monocyte derived dendritic cells (DC) are professional antigen presenting cells widely investigated as stimulators of alpha/beta T cells. But only little is known about the interaction of gamma/delta T cells and monocyte derived DC. Here, we investigated the effect of coculture of mature DC unpulsed or pulsed with ibandronate on the proliferation and cytotoxic activity of isolated gamma/delta T cells. After coculturing monocyte derived DC with isolated gamma/delta T cells, proliferation of gamma/delta T cells was enhanced as determined by the (3)H thymidine uptake assay. Also, IFN-gamma secretion was increased after coculture with DC. As DC are well known to induce activation of alpha/beta T cells we investigated whether the cytotoxic activity of gamma/delta T cells could be increased by coculture with DC. We found no difference in cytotoxic activity of gamma/delta T cells alone or cocultured with unpulsed or pulsed mature DC. Also, sensitizing of myeloma cells by addition of ibandronate could not increase lysis by gamma/delta T cells. In conclusion, monocyte derived DC are capable of stimulating proliferation and secretion of IFN-gamma of gamma/delta T cells but do not exert an effect on cytotoxic activity of gamma/delta T cells against myeloma cells. PMID- 15894119 TI - Interferon gamma bound to extracellular matrix changes the hyporesponsiveness to LPS in crypt but not villous intestinal epithelial cells. AB - Intestinal epithelial cells (IEC) are hyporesponsive to LPS. Responsiveness to luminal bacteria has been implicated in the pathogenesis of inflammatory bowel diseases (IBD). In support of this, previous studies have demonstrated that some intestinal epithelial cell lines are induced by IFN-gamma to respond to LPS. However, both the responsiveness to LPS and the effect of IFN-gamma in intestinal cell lines are heterogeneous. In addition, IFN-gamma may be sequestered in the extracellular matrix (ECM) compartment. The ECM-bound form is more effective than soluble IFN-gamma in producing its biological effects in several experimental models. We investigated the effect of ECM-bound and soluble IFN-gamma treatment on interleukin-8 (IL-8) secretion in response to LPS in freshly isolated villous and crypt cells. We demonstrate that ECM-bound, but not soluble IFN-gamma, induced an increase in IL-8 secretion in response to LPS in undifferentiated crypt cells. This effect was associated with an increase in TLR4 expression. In contrast, mature villous cells did not modify their response to LPS when treated with IFN-gamma (ECM-bound or soluble). These results suggest that selective changes in immature crypt cells induced by IFN-gamma bound to extracellular matrix could contribute to inappropriate responsiveness to commensal bacteria in IBD. PMID- 15894120 TI - Adoptive transfer of mast cells does not enhance the impaired survival of Kit(W)/Kit(W-v) mice in a model of low dose intraperitoneal infection with bioluminescent Salmonella typhimurium. AB - Mast cells are important effector cells in IgE-associated immune responses, but also can contribute to host defense in certain examples of bacterial infection. We found that genetically mast cell-deficient WBB6F1-Kit(W)/Kit(W-v) mice exhibited more bacterial CFUs per spleen by 6 days after intraperitoneal injection of bioluminescent Salmonella typhimurium, and died more rapidly after infection, than did the congenic WBB6F1-Kit(+/+) wild type mice. Adoptive transfer of bone marrow-derived cultured mast cells of Kit(+/+) origin to the peritoneal cavity of Kit(W)/Kit(W-v) mice resulted in engraftment of mast cells in the peritoneal cavity and mesentery of the recipient mice, and the development of large numbers of mast cells in the spleen. However, such mast cell-engrafted Kit(W)/Kit(W-v) mice appeared sicker after intraperitoneal injection with S. typhimurium than did mast cell-deficient Kit(W)/Kit(W-v) mice, and exhibited numbers of CFUs of bacteria per spleen, and a survival curve, that were not significantly different than those of Kit(W)/Kit(W-v) mice. These results, when taken together with prior studies investigating the roles of mast cells in innate immunity, strongly suggest that whether mast cells can be shown to have a significant role in enhancing survival during bacterial infections may depend critically on the details of the particular experimental systems examined. PMID- 15894121 TI - Mitogen-activated protein kinase pathway controls autoimmune regulator (AIRE) gene expression in granulo-monocyte colony stimulating factor (GM-CSF)-stimulated myelomonocytic leukemia OTC-4 cells. AB - Autoimmune regulator (AIRE) gene is a responsible gene for the rare autosomal recessive autoimmune disease: autoimmune-polyendocrinopathy-candidiasis ectodermal dystrophy (APECED). Although it has been reported that AIRE is expressed in the thymic epithelial cells and monocyte-dendritic cell lineage, the regulatory mechanisms of AIRE gene expression have as yet been poorly understood. Here we show that the expression of AIRE gene was induced in granulo-monocyte colony stimulating factor (GM-CSF)-stimulated myelomonocytic leukemia OTC-4 cells. In GM-CSF-stimulated OTC-4 cells, stat5 was not phosphorylated, while mitogen-activated protein kinases (MAPKs), including MAPK kinase (MEK) 1/2 and p38 MAPK, were phosphorylated, indicating activation of MAPK pathway. In addition, the expression of AIRE gene was inhibited by specific p38 MAPK inhibitor (SB203580), whereas the expression was rather enhanced by the MEK1/2 inhibitor (U0126), suggesting that AIRE gene expression is regulated by mitogen activated protein kinase pathway. PMID- 15894122 TI - Lack of relationship between interleukin-6 and CRP levels in healthy male athletes. AB - Interleukin-6 (IL-6) is the main cytokine involved in the induction of acute phase response, which includes synthesis of certain proteins in the liver, one of which is C-reactive protein (CRP). The aim of this study was to assess the impact of IL-6 released during physical exercise on CRP generation in healthy male athletes. Fourteen young cyclists were enrolled in the study, which involves the performance of strenuous physical exercise. Serum levels of IL-6 and CRP were measured at rest before exercise, and immediately after and 2h after cessation of exercise. IL-6 level was increased 2.42-fold immediately after, and 21.67-fold 2h after exercise. Serum CRP level did not change significantly over the course of observation: it was 3.25 mg/dl before, 2.36 mg/dl immediately after and 2.71 mg/dl 2h after exercise and unrelated to IL-6 level. No correlation between serum levels of IL-6 and CRP was observed during the period of observation. We conclude that under certain circumstances, acute, pulsatile release of IL-6 does not stimulate synthesis of CRP. PMID- 15894123 TI - Antioxidant approach to disease management and the role of 'Rasayana' herbs of Ayurveda. AB - The disease preventive and health promotive approach of 'Ayurveda', which takes into consideration the whole body, mind and spirit while dealing with the maintenance of health, promotion of health and treating ailments is holistic and finds increasing acceptability in many regions of the world. Ancient Ayurvedic physicians had developed certain dietary and therapeutic measures to arrest/delay ageing and rejuvenating whole functional dynamics of the body system. This revitalization and rejuvenation is known as the 'Rasayan chikitsa' (rejuvenation therapy). Traditionally, Rasayana drugs are used against a plethora of seemingly diverse disorders with no pathophysiological connections according to modern medicine. Though, this group of plants generally possesses strong antioxidant activity, only a few have been investigated in detail. Over about 100 disorders like rheumatoid arthritis, hemorrhagic shock, CVS disorders, cystic fibrosis, metabolic disorders, neurodegenerative diseases, gastrointestinal ulcerogenesis and AIDS have been reported as reactive oxygen species mediated. In this review, the role of free radicals in these diseases has been briefly reviewed. 'Rasayana' plants with potent antioxidant activity have been reviewed for their traditional uses, and mechanism of antioxidant action. Fifteen such plants have been dealt with in detail and some more plants with less work have also been reviewed briefly. PMID- 15894124 TI - Protective effect of Khamira Abresham Uood Mastagiwala against free radical induced damage in focal cerebral ischemia. AB - The effect of Khamira Abresham Uood Mastagiwala (KAUM) (a preparation of Indian System of Unani Medicine) on the activity of antioxidant enzymes, glutathione reductase (GR), glutathione S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT) and the content of glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) was studied in the middle cerebral artery occluded (MCAO) rats after 15 days pretreatment (200 mg/kg body weight (b.wt.), orally) of Khamira Abresham Uood Mastagiwala. The rats were trained and assessed for neurobehavioral activity using Cook's climbing pole. The middle cerebral artery of adult male Wistar rats was occluded for 2 h and reperfused for 22 h. The activity of GPx, GST, GR, catalase and content of GSH was decreased significantly in MCAO group as compared with sham. The rats of MCAO + KAUM group have shown a significant protection in the activity of above-mentioned antioxidant enzymes and content of glutathione when compared with MCAO group. The significantly elevated level of TBARS in MCAO group was depleted significantly by the pretreatment of animals with KAUM in MCAO group. The neurobehavioral assessment has also strengthened the above biochemical data thereby indicating that the therapeutic intervention of KAUM, which is a potent cardiac and melancholic tonic, can be used to prevent or reduce the deterioration caused by free radicals thereby preventing subsequent pathological and biochemical changes which occur during cerebral ischemia. PMID- 15894125 TI - Immunosuppressive properties of an ethyl acetate fraction from Euphorbia royleana. AB - The objective of the study was to investigate the activity of the ethyl acetate (EA) fraction of Euphorbia royleana latex on cellular and humoral-mediated immune responses and phagocytic function of the cells of the reticuloendothelial system in mice. Oral administration of EA at doses of 50, 100 and 200 mg/kg p.o. in mice with sheep red blood cells (SRBC) as an antigen-inhibited both the delayed-type hypersensitivity reaction and the production of circulating antibody titre. Reduction of CD4+ T cell counts in the peripheral whole blood and the neutrophil counts in pleural exudates of the animals treated with EA was observed by flowcytometric analysis. Process of phagocytosis was also inhibited in in vivo and in vitro experimental test models. The oral LD50 in both rats and mice was more than 2.5 g/kg body weight. PMID- 15894126 TI - Antimicrobial properties of Phrygilanthus acutifolius. AB - Ethanol extract of flowers of Phrygilanthus acutifolius (Ruiz & Pav.) Eichler (Loranthaceae) inhibited the growth of both Gram (+) bacteria (Staphylococcus aureus, Staphylococcus saprophyticus and Enterococcus faecalis) and Gram (-) bacteria (Serratia marcescens, Acinetobacter sp. and Pseudomonas aeruginosa). This extract was bactericidal against Staphylococcus aureus and bacteriostatic against Pseudomonas aeruginosa. Morphological evidence suggests that the extract causes the swelling of the bacterial body of Staphylococcus aureus, the disintegration of the cell surface and the cell death. Bactericidal activity was optimal at pH 7.5 and was not affected by different ionic strengths. The presence of Mg2+ in the culture medium of Phrygilanthus acutifolius diminished the sensitivity of Pseudomonas aeruginosa strain against the extract. Test results would tend to corroborate the folk belief that the flowers of this plant are efficacious against respiratory infections and would justify its further investigation. PMID- 15894127 TI - Antidiabetic plants used by Sikkim and Darjeeling Himalayan tribes, India. AB - Sikkim and Darjeeling Himalayan region is characterized by a rich floral diversity and an equally rich ethnomedicinal tradition. Herbal medicine is the dominant system of medicine practiced by the local tribes of this region for the treatment of diabetes. During the course of the present studies it was found that 37 species of plants belonging to 28 families are used as antidiabetic agents in the folk medicinal practices in the region and 81% of these plants are hitherto unreported as hypoglycemic agents. This finding may lead to serious research towards developing new and efficient drugs for diabetes. PMID- 15894128 TI - Preliminary pharmacological studies on Piper chaba stem bark. AB - Piper chaba Hunter (Piperaceae) is a common pepper in the southern part of Bangladesh. Various parts of this plant have been extensively used in different traditional formulations including ayurveda. In order to rationalize the ethnomedical uses of this plant in a number of ailments, the methanol extract of the stem bark was subjected to preliminary evaluation for analgesic, anti inflammatory, diuretic, anti-diarrhoeal, effect on gastrointestinal motility and CNS depressant activity in mice and rat at 125, 250 and 500 mg/kg body weight doses. The extract at given doses significantly and dose dependently reduced the frequency of acetic acid induced writhing in mice, prolonged the tail flicking latency in mice, reduced Carrageenan-induced paw edema volume in rat, delayed the onset as well as reduced the frequency of castor oil induced diarrhoeal episodes in mice, decreased gastrointestinal motility as assessed by the charcoal motility test in mice and prolonged pentobarbitone induced sleeping time in mice. However at the same doses, the extract exhibited moderate diuretic activity only at the highest dose. PMID- 15894129 TI - Ethnoveterinary medicine in the search for antimicrobial agents: antifungal activity of some species of Pterocaulon (Asteraceae). AB - Based on informal interview, ethnoveterinary information about plants used in the treatment of skin diseases were obtained. Plants from the genus Pterocaulon (Asteraceae) known as "quitoco" are used to treat problems popularly diagnosed as "mycoses", which can have both fungic and bacterial etiology. In order to validate this traditional practice, the crude methanolic extracts and fractions from the aerial parts of three species of Pterocaulon (Pterocaulon alopecuroides (Lam.) D.C., Pterocaulon balansae Chodat. and Pterocaulon polystachyum D.C.) grown in southern Brazil were analyzed for the in vitro antifungal activity against a panel of standardized and clinical opportunistic pathogenic yeasts and filamentous fungi including dermatophytes by the agar dilution method. The crude methanolic extract of Pterocaulon alopecuroides was the most active followed by the extract of Pterocaulon polystachyum. Pterocaulon balansae crude methanolic extract was the less active but its lipophilic fractions showed remarkable activity mainly against the dermatophytes. PMID- 15894130 TI - Anti-inflammatory and analgesic properties of water-ethanolic extract from Pothomorphe umbellata (Piperaceae) aerial parts. AB - Anti-inflammatory and analgesic activities as well as the median lethal dose (LD50) of water-ethanolic extract (PHE) of the aerial parts of Pothomorphe umbellata were evaluated in animal models. The ED(50) (oral) for the inhibition of carrageenan-induced rat paw edema assay was determined to be 550 mg/kg, while the LD(50) was higher than 2.0 g/kg. At a dose of 550 mg/kg, PHE inhibited the inflammatory process by 48.7% (P < 0.05) on the third hour of the assay (edema peak) when compared to the untreated control. Indomethacin, the positive control used in this test, inhibited the edema by 58.6% at a dose of 10 mg/kg, when compared to the untreated control (P < 0.05). All three fractions--hexane, methylene chloride and ethyl acetate--obtained by partition of PHE with respective solvents also showed inhibition of the edema induced by carrageenan over a period of 4h but the methylene chloride fraction showed the best activity. The activity shown by the methylene chloride fraction at 200 mg/kg was comparable to that exhibited by indomethacin at a dose of 10mg/kg. The number of writhings induced by a 0.6% acetic acid solution intraperitoneal injection was decreased by 22% (P < 0.05) in the group treated orally with Pothomorphe umbellata crude extract. PHE also inhibited the granulomatous tissue formation in rats by 6.2% (P < 0.05). In the same assay, topically applied dexamethasone decreased the granuloma formation by 14.2%. The above results suggest that Pothomorphe umbellata crude extract has analgesic and anti-inflammatory properties supporting its folkloric use for the treatment of these conditions. PMID- 15894131 TI - Model of inhibition of Thermus aquaticus polymerase and Moloney murine leukemia virus reverse transcriptase by tea polyphenols (+)-catechin and (-) epigallocatechin-3-gallate. AB - Non-nutritional polyphenolic compounds such as (+)-catechin and (-) epigallocatechin-3-gallate (EGCG) are known as anticancer chemopreventive agents and have been utilised for medical purposes in form of tea drinking. Documented anticancer properties of these compounds result from their antioxidant effects. However, also direct alteration of an enzyme performance has been reported and deserves more attention. In this paper, a direct effect of catechin and EGCG on the performance of reverse transcription (RT) and/or polymerase chain reaction (PCR) was studied. Both tea polyphenolic compounds were added into real-time RT PCR reactions and the fluorescence data obtained were fitted with a mathematical model. Several parameters of PCR performance were compared, obtained from the mathematical model for reactions with and without addition of (+)-catechin and EGCG. Addition of EGCG to enzyme reaction seems to inhibit the RT reaction (p < 0.05) and to slow down the DNA polymerase reaction (p < 0.001). Similarly, (+) catechin inhibited the DNA amplification (p < 0.01) but had no effect on the RT reaction. The effects could be observed in physiological flavanol concentrations ranging from 10(-5) to 10(-8)M. PMID- 15894132 TI - Use of Pavo cristatus feather extract for the better management of snakebites: neutralization of inflammatory reactions. AB - In Indian traditional medicine, peacock feather in the form of ash (Bhasma) or water extract are used against snakebite and to treat various problems associated with lungs. This study was aimed to evaluate the water extract of peacock feather (PCF) against the local tissue damage caused due to snakebite. PCF water extract showed inhibition towards phospholipase A2 enzyme activity from snake venom (Naja naja and Vipera russelii), inflammatory fluids (synovial, pleural, ascites) and normal serum in a dose-dependent manner. Hyaluronidase and proteases are other major enzymes in snake venoms responsible for local tissue damage. PCF water extract inhibited hyaluronidase and proteolytic enzyme activities from Vipera russelii, Naja naja and Trimeresurus malabaricus venom. The active principle is a hydrophilic molecule easily extractable in water or polar solvents. PCF water extract gave positive results for the presence of protein and secondary metabolites like carotenoids and steroids. Analysis of metal ions revealed that iron is the major ion (> 20-fold). Other metal ions detected in smaller amount are copper, chromium, zinc and nickel. The least amount of ion detected is gold. Co-injection of PCF water extract with snake venom and inflammatory PLA2 enzymes neutralize the edema inducing activity of all the PLA2 enzymes studied. Since it inhibits hyaluronidase and proteases enzyme activity from snake venom PCF water extract is a powerful neutralizing agent, which has therapeutic application against venom toxicity. PMID- 15894133 TI - Punica granatum flower extract, a potent alpha-glucosidase inhibitor, improves postprandial hyperglycemia in Zucker diabetic fatty rats. AB - Postprandial hyperglycemia plays an important role in the development of type 2 diabetes and has been proposed as an independent risk factor for cardiovascular diseases. The flowering part of Punica granatum Linn. (Punicaceae) (PGF) has been recommended in Unani literature as a remedy for diabetes. We investigated the effect and action mechanism of a methanolic extract from PGF on hyperglycemia in vivo and in vitro. Oral administration of PGF extract markedly lowered plasma glucose levels in non-fasted Zucker diabetic fatty rats (a genetic model of obesity and type 2 diabetes), whereas it had little effect in the fasted animals, suggesting it affected postprandial hyperglycemia in type 2 diabetes. In support of this conclusion the extract was found to markedly inhibit the increase of plasma glucose levels after sucrose loading, but not after glucose loading in mice, and it had no effect on glucose levels in normal mice. In vitro, PGF extract demonstrated a potent inhibitory effect on alpha-glucosidase activity (IC50: 1.8 microg/ml). The inhibition is dependent on the concentration of enzyme and substrate, as well as on the length of pretreatment with the enzyme. These findings strongly suggest that PGF extract improves postprandial hyperglycemia in type 2 diabetes and obesity, at least in part, by inhibiting intestinal alpha glucosidase activity. PMID- 15894134 TI - Effect of bee venom on aromatase expression and activity in leukaemic FLG 29.1 and primary osteoblastic cells. AB - The effect of bee venom aqua-acupuncture (BVA) (api-toxin), a traditional immunosuppressive Korean aqua-acupuncture, on the bone function in human osteoblastic cells was studied. To provide insights into the effect of BVA on aromatase activity in bone-derived cells, we examined the human leukaemic cell line FLG 29.1, which is induced to differentiate toward the osteoclastic phenotype by TPA and TGF-beta1, and the primary first-passage osteoblastic cells (hOB). Southern blot of RT-PCR products with a 32P-labeled cDNA probe for the human aromatase demonstrated that FLG 29.1 and hOB cells express aromatase mRNA. Gene expression and enzyme activity were stimulated in a time-dependent fashion by 5.0 microl/ml BV and by either 1-50 nM TPA or 0.01-0.5 ng/ml TGF-beta1, with maximal responses after 2-3 h exposure. After 24 h incubation of the cells in the absence of these stimuli the aromatase mRNA and the protein were barely detectable. These findings demonstrate that cells of the osteoclastic lineage synthesize aromatase in vitro by the local cytokine of TGF-beta1 and BVA. These can offer an explanation for the lack of development of osteoarthritis in BVA treated patients. PMID- 15894135 TI - Antibacterial properties of common herbal remedies of the southwest. AB - Curanderismo, widely practiced in the southwest, is an alternative medical system that has been neglected by scientific research. This project analyzed the antibiotic properties of 23 common herbal remedies used in South Texas to treat wounds and infections. Ethanolic tinctures and aqueous extracts of each plant were prepared and applied to blank diffusion disks. These disks were desiccated and used in Kirby-Bauer disk diffusion susceptibility tests on three bacteria: Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. Control disks contained solvent only. The efficacy of the tinctures and aqueous extracts was compared to that of commercially prepared antibiotic diffusion disks. No inhibition was observed with the aqueous extracts. The various tincture-saturated disks produced zones of clearance ranging from 1 to 5 mm. Ten plants consistently inhibited bacterial growth of Staphylococcus aureus. None of the plants tested produced consistent inhibition of the two Gram-negative species, Pseudomonas aeruginosa and Escherichia coli. No zones of clearance were produced by the solvent-only control disks. The zones of clearance produced by commercial antibiotics were, on average, larger and more uniform than those produced by the tincture disks. Thus, it appears that some of the herbal remedies used in folk medicine are potentially effective antibacterial agents against Staphylococcus aureus. PMID- 15894136 TI - Antioxidant phenolic constituents from Fagopyrum dibotrys. AB - Fagopyrum dibotrys (D. Don.) Hara. is an erect perennial Polygonaceous herb. In China, its rhizome was used as folk medicine for the treatment of lung diseases, dysentery and rheumatism. The crude aqueous acetone extract from the rhizomes of this plant exhibited high antioxidant activity (SC50 = 10.95 microg/mL) in 1,1 diphenyl-2-picryldydrazyl (DPPH) radical scavenging assay. Detailed chemical investigation on the extract led to the isolation of two new phenols (1 and 2), together with 14 known antioxidant phenolic compounds (3-16). Their structures were determined by detailed spectroscopic analysis. The two new compounds were characterized as 3-methyl-gossypetin 8-O-beta-D-glucopyranoside (1) and 1,3,6' tri-p-coumaroyl-6-feruloyl sucrose (diboside A, 2). The radical scavenging activity of all the isolated compounds was also described. PMID- 15894137 TI - Influence of extracts of Stryphnodendron polyphyllum Mart. and Stryphnodendron obovatum Benth. on the cicatrisation of cutaneous wounds in rats. AB - Stem bark of the two species Stryphnodendron polyphyllum Mart. and Stryphnodendron obovatum Benth., Leguminosae, was investigated for wound healing, antibacterial and antioxidant activity. These plants contain 12 and 19% tannins in their stem bark, respectively, and are widely used in traditional medicine in Brazil. The total content of phenolics of the crude extract (CE) of Stryphnodendron obovatum was 76.95 +/- 2.98% (CV = 3.87%) and of the ethyl acetate fraction (EAF) was 89.13 +/- 0.34% (CV = 0.38%); whereas in Stryphnodendron polyphyllum the CE phenolics content was 51.62+/-1.53% (CV = 2.96%) and the EAF phenolics content was 59.00 +/- 1.91% (CV = 3.24%). The tannin content of CE from Stryphnodendron obovatum [36.58 +/- 0.35% (CV = 0.98%)] was about 11% higher than in CE from Stryphnodendron polyphyllum [25.43 +/- 0.96% (CV = 3.77%)]. The difference between the species was even greater in the EAF: in Stryphnodendron obovatum the EAF phenolics content was 55.01 +/- 0.36% (CV = 0.65%), whereas in Stryphnodendron polyphyllum the content was 36.16 +/- 0.42% (CV = 1.16%). The healing effect of ointments containing 2.5% crude lyophilised extract (PCE) and 2.5% ethyl-acetate lyophilised fraction (PEA) of the stem bark of Stryphnodendron polyphyllum and Stryphnodendron obovatum was studied in cutaneous wounds of Wistar rats after 4, 7 and 10 days of treatment. Epithelial cell proliferation in the area of re-epithelialisation of the wounds was evaluated by counting the metaphases blocked by vincristine sulfate. With PCE an increase in epidermal growth was observed after 4 and 7 days of treatment with Stryphnodendron polyphyllum, and after 7 and 10 days of treatment with Stryphnodendron obovatum. Wounds treated with PEA of Stryphnodendron obovatum showed increased epidermal growth only 4 days after the treatment; for Stryphnodendron polyphyllum, epidermal growth was observed after 4 and 7 days of treatment. Both the CE and the EAF fractions of Stryphnodendron polyphyllum and Stryphnodendron obovatum showed antibacterial activity against Staphylococcus aureus with MIC values of 125 and 250 microg/ml, respectively. Gram-negative bacteria tested were not inhibited by extracts and fractions at concentrations > 1000 microg/ml. The antioxidant activity through reduction of the DPPH radical in TLC, confirmed the anti-radical properties of these extracts in both species. CE and EAF of both species showed a radical scavenging activity (RSA) and protected DPPH from discolouration, already at 0.032 microg/ml. The extract from Stryphnodendron polyphyllum were more effective than those Stryphnodendron obovatum, although the former had a lower tannin content. PMID- 15894138 TI - Ethnobotanical study of some Ghanaian anti-malarial plants. AB - An ethnobotanical study was conducted in the Wechiau Community Hippopotamus Sanctuary area in Ghana, through interviews and quadrate studies, to investigate the range and abundance of species used in the treatment of malaria. Forty-one species belonging to 17 families were encountered during the study. Of the 17 families studied Leguminosae and Anacardiaceae predominated in terms of number of species used to treat malaria. Eight plant species namely, Afraegle paniculata (Rutaceae), Haematostaphis barteri (Anacardiaceae), Indigo era pulchra (Leguminosae), Monanthotaxis sp. (Annonaceae), Ozoroa insignis (Anacardiaceae), Strychnos innocua (Loganiaceae), Strychnos spinosa (Loganiaceae) and Xeroderris stuhlmannii (Leguminosae) have not previously been documented for the treatment of malaria in Ghana. The results are discussed and recommendations made for future research to support the conservation and sustainable harvesting of the species reported to have medicinal properties. PMID- 15894139 TI - Research of enzymatic activities of fresh juice and water infusions from dry herbs. AB - Research was done on the presence of enzymes in juice obtained from fresh plant material from Chamomilla recutita L. (Rauschel)-anthodium, Lamium album L.-flos, Calendula officinalis L.-flos, Plantaginis lanceolata L.-folium and Euphrasiae rostkoviana Hayne-herba, and in the prepared water infusion of these materials; the objective was to determine the activity of enzymes which beside biologically active substances may have an influence of the final therapeutic effect of the applied plant preparations. The research was conducted by means of the API ZYM system (bioMerieux). Higher enzymatic activities were found in fresh juices of the examined plant material than in prepared water infusions from dried plants. In both cases naphthol-AS-BI-phosphohydrolase should have highest activity. The second one in terms of activity out of 17 studied enzymes was acidic phosphatase. The highest enzymatic activity of fresh juice was found in Lamii albi flos and Calendulae officinalis flos. Water infusions showed the highest enzymatic activity in Lamii albi flos, Chamomille recutita anthodium and Plantaginis lanceolata folium. Drying the plant material resulted in decreased enzymatic activities but not in the case of naphthol-AS-BI-phosphohydrolase and acidic phosphatase which showed very low activities. The complex composition of plant materials in terms of content of biologically active substances may imply that the therapeutic effect might be directly related to the quantity and activity of plant enzymes present in preparations applied in therapeutics. PMID- 15894140 TI - Isolation and antimicrobial activity of two phenolic compounds from Pulicaria odora L. AB - The essential oil of Pulicaria odora, a Moroccan medicinal plant; was analyzed by GC-MS, and subjected to column chromatography on silica gel. Two major constituents were isolated and identified as 2-isopropyl-4-methylphenol (1) and isobutyric acid 2-isopropyl-4-methylphenylester (2), by analysis of spectroscopic data (MS, 1H NMR, 13C NMR, DEPT, COSY, HMQC and HMBC experiments). The isolated compounds are reported for the first time from Pulicaria genus. The essential oil and its major constituents (compounds 1 and 2) were examined for antibacterial and antifungal activity in vitro using the diffusion and dilution methods. Results showed that the essential oil and the 2-isopropyl-4-methylphenol (1) exhibited a very significant antibacterial and antifungal activity, while the isobutyric acid 2-isopropyl-4-methylphenylester (2) was inactive for all tested strains. PMID- 15894141 TI - Ultrasound-assisted extraction methodology as a tool to improve the antioxidant properties of herbal drug Xiao-chia-hu-tang. AB - Xiao-chai-hu-tang (XCHT) is an important Chinese herbal prescription for curing many types of liver diseases. The contents of bioactive constituents (saikosaponins a, c and d, baicalin, baicalein, and glycyrrhizic acid), and antioxidant properties of XCHT extracts prepared with ultrasound-assisted (US) extraction in combination with ethanol (up to 95%) as extraction modifier were studied. The results showed that the US extraction significantly increased the bioactive constituents concentrations and antioxidant properties of XCHT extracts when compared with the XCHT prepared with traditional boiling-water extraction. Among the XCHT extracts made with US extraction, the sample prepared with 95% ethanol showed the highest bioactive constituent concentrations and the best antioxidant functionalities. The results suggest that US extraction of XCHT is feasible to replace the traditional time-consuming and low efficiency preparation procedure in the future modernized and commercialized manufacture of this highly valuable Chinese herbal medicine. PMID- 15894142 TI - Antifungal activities of six South African Terminalia species (Combretaceae). AB - A serial microplate dilution method developed for bacteria was modified slightly and gave good results with several fungi. The antifungal activity of acetone, hexane, dichloromethane and methanol leaf extracts of six Terminalia species (Terminalia prunioides, Terminalia brachystemma, Terminalia sericea, Terminalia gazensis, Terminalia mollis and Terminalia sambesiaca) were tested against five fungal animal pathogens (Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, Microsporum canis and Sporothrix schenkii). Methanol extracted the highest quantity, but the acetone extracts had the highest antifungal activity. Some of the extracts had antioxidant activity. Most of the antifungal extracts had MIC values of c. 0.08 mg/ml, some were with MIC values as low as 0.02 mg/ml. Microsporum canis was the most susceptible microorganism and Terminalia sericea extracts were the most active against nearly all microorganisms tested. PMID- 15894143 TI - Antibacterial screening of some Peruvian medicinal plants used in Calleria District. AB - Nine ethanol extracts of Brunfelsia grandiflora (Solanaceae), Caesalpinia spinosa (Caesalpiniaceae), Dracontium loretense (Araceae), Equisetum giganteum (Equisetaceae), Maytenus macrocarpa (Celastraceae), Phyllanthus amarus (Euphorbiaceae), Piper aduncum (Piperaceae), Terminalia catappa (Combretaceae), and Uncaria tomentosa (Rubiaceae), medicinal plants traditionally used in Calleria District for treating conditions likely to be associated with microorganisms, were screened for antimicrobial activity against nine bacterial strains using the broth microdilution method. Among the plants tested, Phyllanthus amarus and Terminalia catappa showed the most promising antibacterial properties, inhibiting all of the strains tested with minimum inhibitory concentrations (MICs) ranging from 0.25 to 16 mg/ml. The extract from aerial part of Piper aduncum was significantly more active against Gram-positive (MICs ranging from 1 to 2 mg/ml) than against Gram-negative bacteria (MICs > 16 mg/ml). PMID- 15894144 TI - Assessing the burden of disease in the United States using disability-adjusted life years. AB - OBJECTIVES: Burden of disease studies have been implemented in many countries using the disability-adjusted life year (DALY) to assess major health problems. METHODS: We applied methods developed by the World Bank and World Health Organization (WHO) to data specific to the United States to compute DALYs. We compared the results of this analysis to international estimates published by WHO for developed and developing regions of the world. RESULTS: In the mid-1990s, the leading sources of premature death and disability in the United States, as measured by DALYs, were cardiovascular conditions, breast and lung cancers, depression, osteoarthritis, diabetes mellitus, and alcohol use and abuse. In addition, motor vehicle-related injuries and the HIV epidemic exacted a substantial toll on the health status of the U.S. population, particularly among racial/ethnic minorities. The major sources of death and disability in these latter populations were more similar to patterns of burden in developing rather than developed countries. CONCLUSIONS: This analysis provides the first detailed, comprehensive estimates using DALYs of the fatal and nonfatal conditions that exact large health burdens in the United States. PMID- 15894145 TI - Prediction of adult-onset smoking initiation among U.S. Air force recruits using the pierce susceptibility questionnaire. AB - BACKGROUND: At one time, it was expected that if an individual reached young adulthood without smoking, his/her risk for initiation was minimal. However, recent data suggest that smoking initiation among young adults may be increasing. One particularly vulnerable population of young adults for smoking initiation is military recruits. This study examined whether a modified version of the Pierce Susceptibility Questionnaire (PSQ) might be useful for identifying military recruits who are vulnerable for the initiation of tobacco use. METHODS: Active duty recruits entering the U.S. Air Force over a 1-year period (n=31,107) were assessed upon entry into basic military training (BMT), and a subsample was also assessed at 12 months following training. RESULTS: The modified PSQ predicted initiation of cigarette use for all categories of baseline smokers (never, experimenters, former smokers, and regular users). The likelihood of initiating smoking after BMT was substantially higher among those identified as susceptible than among those identified as not susceptible. CONCLUSIONS: It is likely that the PSQ would prove to be a valuable tool in attempts to reduce tobacco use in the military. The predictive abilities of the measure could be used to target intervention and prevention efforts towards those most at risk. PMID- 15894146 TI - Long-term consequences of childhood sexual abuse by gender of victim. AB - BACKGROUND: Childhood sexual abuse (CSA) is a worldwide problem. Although most studies on the long-term consequences of CSA have focused on women, sexual abuse of both boys and girls is common. Thus, a comparison of the long-term effects of CSA by gender of the victim will provide perspective on the need for future research, prevention activities, and treatment of survivors. METHODS: A retrospective cohort study was conducted from 1995 to 1997 among 17,337 adult HMO members in San Diego, California. Participants completed a survey about abuse or household dysfunction during childhood, and multiple other health-related issues. Multivariate logistic regression was used to examine the relationships between severity of CSA (intercourse vs no intercourse) and long-term health and social problems (substance use and abuse, mental illness, and current problems with marriage and family) by gender of victim. Models controlled for exposure to other forms of adverse childhood experiences that co-occur with CSA. Among men, the relationship between the gender of the CSA perpetrator to the outcomes was also examined. RESULTS: Contact CSA was reported by 16% of males and 25% of females. Men reported female perpetration of CSA nearly 40% of the time, and women reported female perpetration of CSA 6% of the time. CSA significantly increased the risk of the outcomes. The magnitude of the increase was similar for men and women. For example, compared to reporting no sexual abuse, a history of suicide attempt was more than twice as likely among both men and women who experienced CSA (p<0.05). Compared with those who did not report CSA, men and women exposed to CSA were at a 40% increased risk of marrying an alcoholic, and a 40% to 50% increased risk of reporting current problems with their marriage (p<0.05). CONCLUSIONS: In this cohort of adult HMO members, experiencing CSA was common among both men and women. The long-term impact of CSA on multiple health and social problems was similar for both men and women. These findings strongly indicate that boys and girls are vulnerable to this form of childhood maltreatment; the similarity in the likelihood for multiple behavioral, mental, and social outcomes among men and women suggests the need to identify and treat all adults affected by CSA. PMID- 15894147 TI - Colorectal cancer screening attitudes and practices preferences for decision making. AB - BACKGROUND: The availability of several effective screening options for colorectal cancer (CRC) screening calls for involving patients in decision making about CRC screening. The current study examined (1) participant characteristics associated with their preferences for participation in CRC screening decision making, (2) correspondence between participant preferences for decision making and their usual participation in decision making, and (3) associations between participant decision-making preferences and CRC screening practices and attitudes. METHODS: Data were obtained using a random, population-based telephone survey, conducted during August 2001 and April 2002, of 2119 community-living adults aged 50 to 75 years (56% female) residing in Long Island, NY. RESULTS: Overall, 77% reported that preferences for CRC screening decision making matched how screening decisions were usually made (simple kappa coefficient=0.67 [0.64 0.69]). Fifteen percent preferred to make screening decisions themselves, while 25% preferred to make decisions after considering their physician's opinion; nearly 50% preferred to share decision making, and 16% preferred that their physician make all screening decisions. Less education was associated with preferring that the physician make all screening decisions. Preferring physician involvement in screening decision making was associated with greater odds of citing no physician recommendation as a barrier to CRC screening, when compared to those who preferred no physician involvement. Preferring no physician involvement in decision making was associated with lower odds of reporting a recent CRC screening exam, as well as lower odds of endorsing positive attitudes and greater odds of endorsing negative attitudes toward CRC screening, when compared to participants who preferred physician involvement in decision making. Their attitudes also reflected intentions not to screen for CRC if they were asymptomatic, as well as the perception that they were not at personal risk for CRC. CONCLUSIONS: Several factors were identified as significantly associated with preferences for decision making and deserve further exploration for their application to clinical practice. PMID- 15894148 TI - Adolescent patterns of physical activity differences by gender, day, and time of day. AB - BACKGROUND: More information about the physical activity of adolescents is needed. This study used objective measurement to investigate differences in activity patterns related to gender, body mass index (BMI), day, and time of day. METHODS: Eighth-grade adolescents (37 boys, 44 girls) wore the Manufacturing Technologies Inc. (MTI) accelerometer for 4 days and kept a previous-day physical activity recall diary in the fall of 2002. Minutes per hour in sedentary, light, and moderate/vigorous activity, as recorded by the MTI, and in nine activity categories, as recorded by the diary, were calculated for three time periods (6:00 am to 2:59 pm, 3:00 pm to 6:59 pm, 7:00 pm to midnight) on each day (Thursday through Sunday). RESULTS: Doubly multivariate analysis of variance revealed significant gender by day by time differences in sedentary (p =0.005) and moderate/vigorous (p <0.001) activity, but no significant BMI interactions. Except on Sunday, boys were less sedentary and more active than girls during the late afternoon period. Significant gender by category (p <0.001) and day by category (p <0.001) interactions were also found in the log data. Boys spent more time engaged in TV/electronics and sports, while girls spent more time in personal care. Three activity categories (sports, social interaction, active transportation) stayed at consistent levels across days, while others varied widely by day of the week. CONCLUSIONS: Except on Sunday, consistent gender differences were found in activity levels, especially for the late afternoon period. Significant increases in sitting, TV/electronic games, and chores were seen for weekend days. Results support strategies to reduce sitting and electronic recreation, which may increase physical activity. PMID- 15894149 TI - Exercise effects on bone mineral density relationships to changes in fitness and fatness. AB - BACKGROUND: Weight loss typically reduces bone mineral density (BMD). Exercise may preserve or increase BMD even while reducing fatness. We examined the relationships among exercise-induced changes in fitness and fatness with BMD. DESIGN: Randomized controlled trial conducted between July 1999 and November 2003. PARTICIPANTS: Men and women (n =115) aged 55 to 75 years. INTERVENTION: Six months of exercise training. MAIN OUTCOME MEASURES: Fitness measured as peak oxygen uptake and muscle strength, body composition by anthropometry, dual-energy x-ray absorptiometry, and magnetic resonance imaging. RESULTS: A total of 51 men and 53 women completed the trial. Exercise increased aerobic and strength fitness and lean body mass, and reduced general and abdominal obesity. BMD did not change among men in either group. Among women exercisers, there were reductions in total skeleton BMD (p =0.02) and greater trochanter BMD (p =0.02). By bivariate correlation, among women, increased femoral neck BMD was associated with increased aerobic fitness (p =0.01) and with reduced body weight (p =0.02) and BMI (p =0.02). In the final regression model, 13% of the change in femoral neck BMD was explained by the change in aerobic fitness (p <0.01). Among the men, increased total hip BMD and femoral shaft BMD were associated with increased lean mass and lower-body strength. In the final regression models, the change in lean mass explained 9% of the variance in total hip BMD (p =0.04). The change in lean mass explained 20% of the change in femoral shaft BMD (p <0.01), and the change in lower-body strength explained an additional 6% (p <0.04). CONCLUSIONS: When examined by group assignment, 6 months of exercise had no effect on BMD among men, and reduced BMD among women. When examined by change in fitness and fatness, women who had the greatest increases in aerobic capacity and men who had the greatest increases in strength and lean mass were more likely to increase their BMD. Exercise-induced reductions in fatness did not lead to bone loss. PMID- 15894150 TI - Objective measures of neighborhood environment and physical activity in older women. AB - BACKGROUND: Regular physical activity is known to help prevent chronic disease and promote healthy aging. Yet, most older women are not regularly active. This study attempts to identify objectively measured attributes of the neighborhood environment that may be associated with physical activity levels in older women. METHODS: Sociodemographics and physical activity level, as measured by pedometer, were assessed in 158 overweight Caucasian and African-American postmenopausal women from southwestern Pennsylvania at the baseline evaluation of a randomized clinical trial in 2002-2003. Geographic information systems technology was used to obtain neighborhood-level data, including neighborhood socioeconomic status (SES) indicators, the median year that homes were built (as a proxy measure for urban form), and proximity to businesses and facilities. Multiple linear regression was used to test associations between individuals' physical activity level and neighborhood characteristics. RESULTS: After controlling for individual age, race/ethnicity, education, smoking status, and body mass index, indicators of low neighborhood SES, living in a neighborhood with homes built between 1950 and 1969 (representing an urban form that is more pedestrian-friendly than after 1969), and living within walking distance (1500 m) of specific types of businesses and facilities were positively associated with individuals' physical activity level measured by pedometer (p <0.05). CONCLUSIONS: Results suggest that certain aspects of the neighborhood environment may have an important influence on the physical activity levels of postmenopausal women. Results warrant future research to clarify the role of these environmental attributes in other populations. PMID- 15894151 TI - Test-retest reliability of adult surveillance measures for physical activity and inactivity. AB - BACKGROUND: Several physical activity measures used in U.S. surveillance systems lack estimates of reliability in this country. This information is needed among diverse populations of women and men, to aid in interpretation and use of the measures. The objective of this study was to document the test-retest reliability of several measures of physical activity and inactivity used in surveillance in a diverse adult population. METHODS: Test and retest surveys were conducted over the telephone with 106 African-American and white women and men living in Forsyth County, North Carolina or Jackson, Mississippi in 2003. Physical activity and inactivity were self-reported using surveillance measures, such as from the Behavioral Risk Factor Surveillance System. Reliability was determined using kappa and intraclass correlation coefficients (ICCs) overall and separately by gender and race. RESULTS: Thirteen percent of the participants met recommendations for physical activity, 44% were insufficiently active, and 43% were inactive. Reliability of the measures to categorize participants into these categories was 0.44 (95% confidence interval [CI]=0.27-0.58). The reliability of several categoric definitions of leisure activity ranged from 0.46 to 0.68. Occupational activity had substantial reliability (0.82, 95% CI=0.72-0.89), while any transportation activity (0.27, 95% CI=0.09-0.44) and walking (0.40, 95% CI=0.23-0.55) were lower. Indicators of inactivity categorized at >7 hours/week included time per week on the computer (0.83, 95% CI=0.57-0.78) and time per week watching television (0.40, 95% CI=0.22-0.54). Some gender and racial differences were noted in the reliability estimates. CONCLUSIONS: In conclusion, this study provides estimates of test-retest reliability for several physical activity and inactivity measures used for surveillance purposes. Validity data, coupled with the reliability estimates reported here and elsewhere, can aid in interpretation and use of these measures in surveillance, as well as in epidemiologic studies. PMID- 15894152 TI - Colorectal cancer screening among African-American and white male veterans. AB - BACKGROUND: Population-based studies from Medicare and privately insured individuals have consistently identified lower rates of colorectal cancer screening tests among African-American versus white individuals. The purpose of this study was to evaluate whether, at a Veterans Affairs (VA) medical center, similar racial/ethnic differences in colorectal cancer screening could be identified. METHODS: Study participants were male veterans, aged > or =50, attending a general medicine clinic in a VA hospital, who had not had either a fecal occult blood test (FOBT) within the past year or a flexible sigmoidoscopy/colonoscopy within the past 5 years. Based on review of electronic medical records, rates of physician recommendation for FOBT, flexible sigmoidoscopy, or colonoscopy, and patient completion of these tests were obtained and compared by race/ethnicity. RESULTS: Sixty percent of 1599 veterans had not undergone recent colorectal cancer screening. Physicians recommended colorectal screening tests equally among African-American and white patients (71.0% vs 68.2%, p=0.44). African-American patients were 1.3 times more likely than white patients to receive colorectal screening procedures (36.3% vs 28.9%, p=0.03). CONCLUSIONS: In contrast to other settings, in a general medicine clinic at a VA hospital, rates of colorectal cancer-screening tests were not lower for African-American patients compared to white patients. PMID- 15894153 TI - Keeping up with evidence a new system for WHO's evidence-based family planning guidance. AB - The World Health Organization (WHO) is responsible for providing evidence-based family planning guidance for use worldwide. WHO currently has two such guidelines, Medical Eligibility Criteria for Contraceptive Use and Selected Practice Recommendations for Contraceptive Use, which are widely used globally and often incorporated into national family planning standards and guidelines. To ensure that these guidelines remain up-to-date, WHO, in collaboration with the Centers for Disease Control and Prevention and the Information and Knowledge for Optimal Health (INFO) Project at the Johns Hopkins Bloomberg School of Public Health's Center for Communication Programs, has developed the Continuous Identification of Research Evidence (CIRE) system to identify, synthesize, and evaluate new scientific evidence as it becomes available. The CIRE system identifies new evidence that is relevant to current WHO family planning recommendations through ongoing review of the input to the POPulation information onLINE (POPLINE) database. Using the Meta-Analysis of Observational Studies in Epidemiology guidelines and standardized abstract forms, systematic reviews are conducted, peer-reviewed, and sent to WHO for further action. Since the system began in October 2002, 90 relevant new articles have been identified, leading to 43 systematic reviews, which were used during the 2003-2004 revisions of WHO's family planning guidelines. The partnership developed to create and manage the CIRE system has pooled existing resources; scaled up the methodology for evaluating and synthesizing evidence, including a peer-review process; and provided WHO with finger-on-the-pulse capability to ensure that its family planning guidelines remain up-to-date and based on the best available evidence. PMID- 15894154 TI - The multiple layers of prevention science research. AB - A systematic framework for prevention science clinical research is described that explicitly links the identification of prevention needs to the implementation of research results in public health programs. The framework integrates five elements: advocacy and policy, community participation, prevention research, acceptability research, and operations and program development. These five elements are often viewed as disparate agendas, but an effective prevention science research enterprise requires that they be coordinated and integrated through all research stages--from the conceptual, to the experimental, and ultimately to the applied. Examples are provided of how this integration can be achieved from our experience in reproductive health and HIV prevention. PMID- 15894155 TI - In response to the 2005, Vol. 28, Issue 1, article entitled public perceptions of childhood obesity. PMID- 15894156 TI - With both eyes open: The Guide to Community Preventive Services. PMID- 15894157 TI - Evidence-based public health policy and practice: promises and limits. PMID- 15894158 TI - Recommendations to improve targeted vaccination coverage among high-risk adults. PMID- 15894159 TI - Methods for conducting systematic reviews of targeted vaccination strategies for the guide to community preventive services. PMID- 15894160 TI - Interventions to improve influenza, pneumococcal polysaccharide, and hepatitis B vaccination coverage among high-risk adults: a systematic review. PMID- 15894161 TI - Effectiveness of designated driver programs for reducing alcohol-impaired driving: a systematic review. PMID- 15894162 TI - Effectiveness of school-based programs for reducing drinking and driving and riding with drinking drivers: a systematic review. PMID- 15894164 TI - Involvement of extracellular Ca2+ influx through voltage-independent Ca2+ channels in endothelin-1 function. AB - This article reviews the types and roles of voltage-independent Ca(2+) channels involved in the endothelin-1 (ET-1)-induced functional responses such as vascular contraction, cell proliferation, and intracellular Ca(2+)-dependent signaling pathways and discusses the molecular mechanisms for the activation of voltage independent Ca(2+) channels by ET-1. ET-1 activates some types of voltage independent Ca(2+) channels, such as Ca(2+)-permeable nonselective cation channels (NSCCs) and store-operated Ca(2+) channels (SOCC). Extracellular Ca(2+) influx through these voltage-independent Ca(2+) channels plays essential roles in ET-1-induced vascular contraction, cell proliferation, activation of epidermal growth factor receptor tyrosine kinase, regulation of proline-rich tyrosine kinase, and release of arachidonic acid. The experiments using various constructs of endothelin receptors reveal the importance of G(q) and G(12) families in activation of these Ca(2+) channels by ET-1. These findings provide a potential therapeutic mechanism of a functional interrelationship between G(q)/G(12) proteins and voltage-independent Ca(2+) channels in the pathophysiology of ET-1, such as in chronic heart failure, hypertension, and cerebral vasospasm. PMID- 15894165 TI - Pathophysiological roles of G-protein-coupled receptor kinases. AB - G-protein-coupled receptor kinases (GRKs) interact with the agonist-activated form of G-protein-coupled receptors (GPCRs) to effect receptor phosphorylation and to initiate profound impairment of receptor signalling, or desensitization. GPCRs form the largest family of cell surface receptors known and defects in GRK function have the potential consequence to affect GPCR-stimulated biological responses in many pathological situations. This review focuses on the physiological role of GRKs revealed by genetically modified animals but also develops the involvement of GRKs in human diseases as, Oguchi disease, heart failure, hypertension or rhumatoid arthritis. Furthermore, the regulation of GRK levels in opiate addiction, cancers, psychiatric diseases, cystic fibrosis and cardiac diseases is discussed. Both transgenic mice and human pathologies have demonstrated the importance of GRKs in the signalling pathways of rhodopsin, beta adrenergic and dopamine-1 receptors. The modulation of GRK activity in animal models of cardiac diseases can be effective to restore cardiac function in heart failure and opens a novel therapeutic strategy in diseases with GPCR dysregulation. PMID- 15894166 TI - IB1/JIP-1 controls JNK activation and increased during prostatic LNCaP cells neuroendocrine differentiation. AB - The scaffold protein Islet-Brain1/c-Jun amino-terminal kinase Interacting Protein 1 (IB1/JIP-1) is a modulator of the c-Jun N-terminal kinase (JNK) activity, which has been implicated in pleiotrophic cellular functions including cell differentiation, division, and death. In this study, we described the presence of IB1/JIP-1 in epithelium of the rat prostate as well as in the human prostatic LNCaP cells. We investigated the functional role of IB1/JIP-1 in LNCaP cells exposed to the proapoptotic agent N-(4-hydroxyphenyl)retinamide (4-HPR) which induced a reduction of IB1/JIP-1 content and a concomittant increase in JNK activity. Conversely, IB1/JIP-1 overexpression using a viral gene transfer prevented the JNK activation and the 4-HPR-induced apoptosis was blunted. In prostatic adenocarcinoma cells, the neuroendocrine (NE) phenotype acquisition is associated with tumor progression and androgen independence. During NE transdifferentiation of LNCaP cells, IB1/JIP-1 levels were increased. This regulated expression of IB1/JIP-1 is secondary to a loss of the neuronal transcriptional repressor neuron restrictive silencing factor (NRSF/REST) function which is known to repress IB1/JIP-1. Together, these results indicated that IB1/JIP-1 participates to the neuronal phenotype of the human LNCaP cells and is a regulator of JNK signaling pathway. PMID- 15894167 TI - Macrophage colony-stimulating factor receptor induces tyrosine phosphorylation of SKAP55R adaptor and its association with actin. AB - The production, survival, and function of monocytes and macrophages are regulated by the macrophage colony-stimulating factor (M-CSF or CSF-1) through its tyrosine kinase receptor. M-CSF receptor activates multiple cytoplasmic pathways in which adaptor and scaffolding proteins play a central role. In this study, we showed that SKAP55-related (SKAP55R) adaptor protein is expressed in myeloid cells and macrophages and is rapidly and transiently tyrosine-phosphorylated in response to M-CSF. M-CSF induced SKAP55R association with other tyrosine-phosphorylated proteins and with actin. When overexpressed in myeloid cells, SKAP55R decreased M CSF-dependent proliferation without affecting differentiation. Altogether, these results demonstrate that SKAP55R adaptor is implicated in the M-CSF signaling pathway and suggest its role as a negative regulator of growth. Moreover, specific association between SKAP55R and actin support the idea that SKAP55R is implicated in the regulation of actin dynamics under the control of M-CSF. PMID- 15894168 TI - Tyrosine phosphatase PTP1B interacts with TRPV6 in vivo and plays a role in TRPV6 mediated calcium influx in HEK293 cells. AB - This study investigates the role of tyrosine phosphorylation and dephosphorylation in the regulation of the Ca(2+) permeant TRPV6 channel. HEK293 cells co-transfected with TRPV6 and the tyrosine phosphatase PTP1B show a constitutive Ca(2+) entry which was independent of tyrosine phosphorylation under resting conditions. Following depletion of intracellular Ca(2+) stores, TRPV6 mediated Ca(2+) entry could be increased in the presence of a tyrosine phosphatase inhibitor (bis-(N,N-dimethyl-hydroxamido) hydroxo-vanadate; DMHV). Inhibition of Src-kinases completely abolished DMHV-induced increase in TRPV6 mediated Ca(2+) influx. Co-transfection with Src led to tyrosine phosphorylation of TRPV6 which could be dephosphorylated by PTP1B. In vivo interaction of TRPV6 with PTP1B was visualized using the bimolecular fluorescence complementation (BiFC) method and proved by co-immunoprecipitation of both proteins. These data indicate that tyrosine phosphorylation is involved in the regulation of the TRPV6 channel protein. PMID- 15894169 TI - CD44v6 promotes proliferation by persisting activation of MAP kinases. AB - CD44v6 is transiently expressed during T cell activation, and constitutively CD44v4-v7 expressing transgenic T cells show accelerated responses towards nominal antigens. The underlying mechanism is unknown. The mouse thymoma EL4 was transfected with CD44 standard isoform (CD44s) or CD44v6 cDNA (EL4-s, EL4-v6). Only EL4-v6 cells proliferated at an over 10-fold higher rate than untransfected cells, displayed up-regulated expression of CD69, CD25, and IL-2, and were protected from apoptosis by CD44v6 cross-linking. In the absence of any stimulus, ERK1/2 was partly phosphorylated, and phosphorylation was significantly increased by CD44v6 cross-linking. The same accounted for JNK, c-jun, and IkappaBalpha. Moreover, NF-kappaB was partly translocated into the nucleus. Instead, CD44s cross-linking induced ERK1/2, JNK, c-jun, and IkappaBalpha phosphorylation only in the context of TCR engagement. No selectively CD44v6 associated transmembrane proteins were uncovered in EL4 cells. However, CD44v6, as opposed to CD44s, did not colocalise with the TCR/CD3 complex after CD3 cross-linking. Furthermore, a CD44-associated 85-kDa protein became hypophosphorylated only after CD44v6 cross linking. Threonine hypophosphorylation of this protein coincided with the activation of MAP and SAP kinases, which was prohibited in the presence of a phosphatase inhibitor. Thus, CD44v6, distinct to CD44s, stimulates autonomously growth and IL-2 secretion of a thymoma line and rescues cells from apoptosis. PMID- 15894170 TI - Activation of phosphoinositide 3-kinase in response to inflammation and nitric oxide leads to the up-regulation of cyclooxygenase-2 expression and subsequent cell proliferation in mesangial cells. AB - In this study, we showed that nitric oxide (NO) donors induced the mesangial cell proliferation and cyclooxygenase-2 (COX-2) protein expression in murine mesangial cells. An inflammatory condition [lipopolysaccharide (LPS) plus interferon-gamma (IFN-gamma)] could also induce cell proliferation and significantly enhance inducible nitric oxide synthase (iNOS) and COX-2 expression. Phosphoinositide 3 kinase (PI3K) inhibitor, LY294002, inhibited these responses. LPS/IFN-gamma induced COX-2 expression in mesangial cells could be inhibited by iNOS inhibitor, aminoguanidine. Selective COX-2 inhibitor, NS398, was capable of inhibiting NO donor- or LPS/IFN-gamma-induced mesangial cell proliferation. Both NO donor and LPS/IFN-gamma markedly activated the PI3K activity and the phosphorylation of Akt and nuclear factor (NF)-kappaB DNA binding activity in mesangial cells, which could be inhibited by LY294002 and transfection of dominant-negative vectors of PI3K/p85 and Akt. These results indicate that a PI3K/Akt-dependent pathway involved in the NO-regulated COX-2 expression and cell proliferation in mesangial cells under inflammatory condition. PMID- 15894171 TI - Direct association of hepatopoietin with thioredoxin constitutes a redox signal transduction in activation of AP-1/NF-kappaB. AB - It has been demonstrated that growth factors quiescin Q6 family was created by the fusion of the sulfhydryl oxidase fragment of the yeast essential for respiration and vegetative growth (ERV)1 prototype [an orthologue of hepatopoietin (HPO)] and thioredoxin (TRX)/disulfide isomerase domain during evolution. In this paper, our results demonstrated that two components of this composite protein, i.e., HPO and TRX, were involved in the same signal transduction and interacted physically in eukaryocyte. When HPO and TRX were cotransfected into COS7 cells, the activity of activator protein-1 (AP-1) and NF kappaB was evidently enhanced compared with the transfection with HPO or TRX alone, at the same time, the phosphorylation of c-Jun was increased. They were colocalized in the cells. By Co-IP and GST pull-down experiments, we found that HPO could physically interact with TRX, which was also confirmed by yeast two hybrid assay. By further investigation, we found both HPO and TRX were sensitive to cellular oxidative state. HPO dimer is in its natural state and could be reduced by dithiothreitol (DTT) in vitro and in vivo. Under the treatment of oxidants such as H(2)O(2) and diamide, the amount of HPO monomer was decreased significantly and assembled into dimer, and the free thiol in TRX was oxidized. HPO could transfer oxidizing equivalents to TRX via direct thiol-disulfide exchange in vitro, the redox state of TRX was also affected by HPO in vivo. Taken together, it was implicated that the oxidizing equivalents might flow from HPO to TRX and then to substrate protein by the dimerization of HPO, and its interaction with TRX finally activates the redox-sensitive transcription factor, suggesting a new redox signal pathway conducted by thiol-disulfide transformation in eukaryocytic cytoplasm. PMID- 15894172 TI - Phosphorylation-independent internalisation and desensitisation of the human sphingosine-1-phosphate receptor S1P3. AB - Here we demonstrate that phosphorylation of the sphingosine-1-phosphate (S1P) receptor S1P(3) is increased specifically in response to S1P. Truncation of the receptor's carboxyl-terminal domain revealed that the presence of a serine-rich stretch of residues between Leu332 and Val352 was essential to observe this effect. Although agonist-occupied wild-type (WT) S1P(3) could be phosphorylated in vitro by G-protein-coupled receptor kinase 2 (GRK2), a role of S1P(3) phosphorylation in controlling S1P(3)-G(q/11) coupling was excluded since A) a phosphorylation-resistant S1P(3) mutant desensitised in a manner indistinguishable from the WT receptor and was phosphorylated to a greater extent than the WT receptor by GRK2 in vitro, and B) co-expression with GRK2 or GRK3 failed to potentiate S1P(3) phosphorylation. S1P(3) phosphorylation was also not required for receptor sequestration away from the cell surface. Together, these data suggest that S1P(3) function is not subject to conventional regulation by GRK phosphorylation and that novel aspects of S1P(3) function distinct from classical G-protein coupling and receptor internalisation may be controlled its carboxyl-terminal domain. PMID- 15894173 TI - Ca2+-independent activation of Bruton's tyrosine kinase is required for store mediated Ca2+ entry in human platelets. AB - Store-mediated Ca(2+) entry (SMCE), which is rapidly activated by depletion of the intracellular Ca(2+) stores, is a major mechanism for Ca(2+) influx. Several studies have involved tyrosine kinases in the activation of SMCE, such as pp60(src), although at present those involved in the early activation steps are unknown. Here we report the involvement of Bruton's tyrosine kinase (Btk) in the early stages of SMCE in human platelets. Cell treatment with thrombin or thapsigargin (TG) plus ionomycin (Iono) results in rapid activation of Btk, which was independent of rise in intracellular Ca(2+) concentration ([Ca(2+)](i)) but dependent on H(2)O(2) generation. Platelet treatment with Btk inhibitors, LFM-A13 or terreic acid, significantly reduced TG+Iono- and thrombin-evoked SMCE. Btk was rapidly activated by addition of low concentrations of H(2)O(2), whose effect on Ca(2+) entry was prevented by Btk inhibitors. Our results indicate that pp60(src) and Btk co-immunoprecipitate after platelet stimulation with TG+Iono, thrombin or H(2)O(2). In addition, we have found that LFM-A13 impaired actin filament reorganization after store depletion and agonist-induced activation of pp60(src), while the inhibitor of pp60(src), a protein that requires actin reorganization for its activation, did not modify Btk activation, suggesting that Btk is upstream of pp60(src). We propose a role for Btk in the early steps of activation of SMCE in human platelets. PMID- 15894174 TI - Heat treatment decreases melanin synthesis via protein phosphatase 2A inactivation. AB - In the present study, we investigated the effects of heat treatment on melanogenesis in a mouse melanocyte cell line (Mel-Ab). It has been reported that activated extracellular signal-regulated kinase (ERK) is responsible for microphthalmia-associated transcription factor (MITF) degradation, which leads to a reduction in tyrosinase protein production and melanin synthesis. Here we demonstrate that heat treatment induces sustained ERK activation, which may inhibit melanogenesis. However, the specific ERK pathway inhibitors, PD98059 or U0126 did not restore heat-induced hypopigmentation. Furthermore, PD98059 or U0126 hardly blocked the heat-induced activation of ERK. These results suggest that heat treatment may inactivate protein phosphatase, and thus ERK activation is maintained. To support this hypothesis, we examined the effects of heat treatment on protein phosphatase 2A (PP2A) activity. The results obtained show that heat treatment inactivates PP2A, which may subsequently cause ERK activation and that heat treatment inhibits MITF promoter activity. Overall, our results demonstrate that heat treatment reduces melanin production in a temperature dependent manner. PMID- 15894175 TI - Merging multimedia presentations and semistructured temporal data: a graph-based model and its application to clinical information. AB - OBJECTIVE: In this paper, we focus on the issue of providing physicians with the capability of representing in a seamless way both temporal aspects of multimedia semistructured data and their temporal presentation requirements. BACKGROUND: Semistructured data are data having some structure, that may be irregular or incomplete and does not necessarily conform to a fixed schema. Semistructured data often contain the description of histories of the considered real world. The eXtensible Markup Language (XML) is becoming a cross compatible and standardized means for representing semistructured clinical data. In the field of medical informatics, there are many ongoing activities concerning XML. In the field of multimedia database systems, the topic related to the integration of several media objects (with their temporal aspects) have been considered both for data modeling and querying issues and for modeling multimedia presentations. METHODOLOGY: We first propose the Multimedia Temporal Graphical Model (MTGM), by representing a clinical database for cardiology patients undergoing cardiac angiographies and then describe it in a formal way. We deal with the problem of expressing MTGM data by XML and of managing MTGM clinical data through an XML based system. We provide both a technique for translating (a part of) an MTGM database into an XML document and some techniques allowing us to obtain presentations defined by means of the Synchronized Multimedia Integration Language (SMIL) from MTGM presentations. RESULTS: MTGM allows one to represent and store clinical information in a semistructured, temporal, and multimedia database. The physician can define multimedia presentations based on the stored data. Multimedia presentations are then stored in the same MTGM database together with temporal clinical information and are thus represented according to the same data model. A prototype based on an XML native database system has been designed and implemented. DISCUSSION AND CONCLUSIONS: In this work we have considered the theoretical and methodological issues concerning the definition of a general data model for describing temporal and multimedia features of semistructured clinical information. Other research and application oriented features, which have not been considered in MTGM, could be investigated for completing MTGM with regard to its applicability to clinical domains: MTGM does not allow one to express times at different levels of granularities, i.e. with different time units, or with indeterminacy; besides the considered valid time, it could be interesting to manage also other temporal dimensions such as the transaction and availability times. Besides being useful for managing multimedia data stored according to widely accepted standards as MPEG and DICOM, nowadays semistructured data, and XML in particular, are becoming the most important way for expressing and exchanging medical knowledge and data: MTGM can be considered as a data model allowing the seamless representation of both (multimedia and temporal) clinical data and knowledge. PMID- 15894176 TI - Predicting breast cancer survivability: a comparison of three data mining methods. AB - OBJECTIVE: The prediction of breast cancer survivability has been a challenging research problem for many researchers. Since the early dates of the related research, much advancement has been recorded in several related fields. For instance, thanks to innovative biomedical technologies, better explanatory prognostic factors are being measured and recorded; thanks to low cost computer hardware and software technologies, high volume better quality data is being collected and stored automatically; and finally thanks to better analytical methods, those voluminous data is being processed effectively and efficiently. Therefore, the main objective of this manuscript is to report on a research project where we took advantage of those available technological advancements to develop prediction models for breast cancer survivability. METHODS AND MATERIAL: We used two popular data mining algorithms (artificial neural networks and decision trees) along with a most commonly used statistical method (logistic regression) to develop the prediction models using a large dataset (more than 200,000 cases). We also used 10-fold cross-validation methods to measure the unbiased estimate of the three prediction models for performance comparison purposes. RESULTS: The results indicated that the decision tree (C5) is the best predictor with 93.6% accuracy on the holdout sample (this prediction accuracy is better than any reported in the literature), artificial neural networks came out to be the second with 91.2% accuracy and the logistic regression models came out to be the worst of the three with 89.2% accuracy. CONCLUSION: The comparative study of multiple prediction models for breast cancer survivability using a large dataset along with a 10-fold cross-validation provided us with an insight into the relative prediction ability of different data mining methods. Using sensitivity analysis on neural network models provided us with the prioritized importance of the prognostic factors used in the study. PMID- 15894177 TI - Evaluation of radiological features for breast tumour classification in clinical screening with machine learning methods. AB - OBJECTIVE: In this work, methods utilizing supervised and unsupervised machine learning are applied to analyze radiologically derived morphological and calculated kinetic tumour features. The features are extracted from dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) time-course data. MATERIAL: The DCE-MRI data of the female breast are obtained within the UK Multicenter Breast Screening Study. The group of patients imaged in this study is selected on the basis of an increased genetic risk for developing breast cancer. METHODS: The k-means clustering and self-organizing maps (SOM) are applied to analyze the signal structure in terms of visualization. We employ k-nearest neighbor classifiers (k-nn), support vector machines (SVM) and decision trees (DT) to classify features using a computer aided diagnosis (CAD) approach. RESULTS: Regarding the unsupervised techniques, clustering according to features indicating benign and malignant characteristics is observed to a limited extend. The supervised approaches classified the data with 74% accuracy (DT) and providing an area under the receiver-operator-characteristics (ROC) curve (AUC) of 0.88 (SVM). CONCLUSION: It was found that contour and wash-out type (WOT) features determined by the radiologists lead to the best SVM classification results. Although a fast signal uptake in early time-point measurements is an important feature for malignant/benign classification of tumours, our results indicate that the wash-out characteristics might be considered as important. PMID- 15894178 TI - Characterization of clustered microcalcifications in digitized mammograms using neural networks and support vector machines. AB - OBJECTIVE: Detection and characterization of microcalcification clusters in mammograms is vital in daily clinical practice. The scope of this work is to present a novel computer-based automated method for the characterization of microcalcification clusters in digitized mammograms. METHODS AND MATERIAL: The proposed method has been implemented in three stages: (a) the cluster detection stage to identify clusters of microcalcifications, (b) the feature extraction stage to compute the important features of each cluster and (c) the classification stage, which provides with the final characterization. In the classification stage, a rule-based system, an artificial neural network (ANN) and a support vector machine (SVM) have been implemented and evaluated using receiver operating characteristic (ROC) analysis. The proposed method was evaluated using the Nijmegen and Mammographic Image Analysis Society (MIAS) mammographic databases. The original feature set was enhanced by the addition of four rule based features. RESULTS AND CONCLUSIONS: In the case of Nijmegen dataset, the performance of the SVM was Az=0.79 and 0.77 for the original and enhanced feature set, respectively, while for the MIAS dataset the corresponding characterization scores were Az=0.81 and 0.80. Utilizing neural network classification methodology, the corresponding performance for the Nijmegen dataset was Az=0.70 and 0.76 while for the MIAS dataset it was Az=0.73 and 0.78. Although the obtained high classification performance can be successfully applied to microcalcification clusters characterization, further studies must be carried out for the clinical evaluation of the system using larger datasets. The use of additional features originating either from the image itself (such as cluster location and orientation) or from the patient data may further improve the diagnostic value of the system. PMID- 15894179 TI - Neural network predictions of significant coronary artery stenosis in men. AB - OBJECTIVE: A neural network system was designed to predict whether coronary arteriography on a given patient would reveal any occurrence of significant coronary stenosis (>50%), a degree of stenosis which often leads to coronary intervention. METHODOLOGY: A dataset of 2004 records from male cardiology patients was derived from a national cardiac catheterization database. The catheterizations selected for analysis from the database were first-time and elective, and they were precipitated by chest pain. Eleven patient variables were used as inputs in an artificial neural network system. The network was trained on the earliest 902 records in the dataset. The next 902 records formed a cross validation file, which was used to optimize the training. A third file composed of the next 100 records facilitated the choice of a cutoff number between 0 and 1. The cutoff number was applied to the last 100 records, which comprised a test file. RESULTS: When a cutoff of 0.25 was compared to the network outputs of all 100 records in the test file, 12 of 46 (specificity=26%) patients without significant stenosis had outputs0.25 (sensitivity=100%). Therefore, the network identified a fraction of the patients in the test file who did not have significant coronary artery stenosis, while at the same time the network identified all of the patients in the test file who had significant stenosis capable of causing chest pain. CONCLUSION: Artificial neural networks may be helpful in reducing unnecessary cardiac catheterizations. PMID- 15894180 TI - A spatio-temporal Bayesian network classifier for understanding visual field deterioration. AB - OBJECTIVE: Progressive loss of the field of vision is characteristic of a number of eye diseases such as glaucoma which is a leading cause of irreversible blindness in the world. Recently, there has been an explosion in the amount of data being stored on patients who suffer from visual deterioration including field test data, retinal image data and patient demographic data. However, there has been relatively little work in modelling the spatial and temporal relationships common to such data. In this paper we introduce a novel method for classifying visual field (VF) data that explicitly models these spatial and temporal relationships. METHODOLOGY: We carry out an analysis of our proposed spatio-temporal Bayesian classifier and compare it to a number of classifiers from the machine learning and statistical communities. These are all tested on two datasets of VF and clinical data. We investigate the receiver operating characteristics curves, the resulting network structures and also make use of existing anatomical knowledge of the eye in order to validate the discovered models. RESULTS: Results are very encouraging showing that our classifiers are comparable to existing statistical models whilst also facilitating the understanding of underlying spatial and temporal relationships within VF data. The results reveal the potential of using such models for knowledge discovery within ophthalmic databases, such as networks reflecting the 'nasal step', an early indicator of the onset of glaucoma. CONCLUSION: The results outlined in this paper pave the way for a substantial program of study involving many other spatial and temporal datasets, including retinal image and clinical data. PMID- 15894181 TI - Drug resistance models for malaria. AB - The main factors affecting the evolution of drug resistance in malaria according to theoretical models are reviewed here. The overwhelming influence on the emergence and rate of spread of drug resistance is the proportion of infected hosts that are treated with drugs. A second important effect is drug efficacy in killing parasites. Factors such as average transmission rate, recombination, the biological cost of resistance, and the mode of gene action also influence the rate of spread but have relatively minor impacts. A simple population dynamics model that captures the epidemiological effects of drug treatment and resistance, as opposed to a population genetics model that does not, is presented in order to illustrate the main conclusions. PMID- 15894182 TI - Genetic manipulation of insects. PMID- 15894183 TI - Ethical, legal and social issues of genetically modifying insect vectors for public health. AB - The use of genetically modified (GM) insects for control of human disease can be consistent with common ethical norms of international society to reduce human suffering. This paper considers a range of ethical issues including animal rights, informed consent, community consensus and environmental viewpoints. Each community needs to decide its own priorities for methodology of disease policy guidance for ethical genetic engineering, and to negotiate with neighbouring countries. The approach to genetically modify insects raises few intrinsic ethical issues; however, important environmental and human health concerns need to be assessed before release of any GM insects. The policy that each community adopts should be the product of open dialogue involving all sectors of society. It can be expected that this process will take years and not all communities will endorse genetic control approaches to insect vectors. PMID- 15894184 TI - Nutritional regulation of vitellogenesis in mosquitoes: implications for anautogeny. AB - Anautogeny is a successful reproductive strategy utilized by many mosquito species and other disease-transmitting arthropod vectors. Developing an understanding of the mechanisms underlying anautogeny in mosquitoes is very important because this reproductive strategy is the driving force behind the transmission of disease to millions of people. Information gained from mosquito studies may also be applicable to other blood feeding insect vectors. The conversion of protein from blood into yolk protein precursors for the developing oocytes is an essential part of the reproductive cycle, and understanding how this process is regulated could lead to safe, specific, and effective ways to block reproduction in blood feeding insects. Great gains have been made in elucidating the mechanisms that regulate vitellogenesis in mosquitoes, especially Ae. aegypti. However, a number of questions remain to be answered to make the picture more complete. In this review, we summarize what is currently known about the nutritional regulation of vitellogenesis in mosquitoes and the questions that remain to be answered about this important biological phenomenon. PMID- 15894185 TI - Mosquito immunity against Plasmodium. AB - Understanding the molecular mechanisms of the innate immune responses of Anopheles gambiae against Plasmodium parasites is of great importance for current efforts to develop novel strategies for malaria disease control. The parasite undergoes substantial stage-specific losses during its development in the mosquito, which in some cases lead to complete refractoriness of the mosquito against the parasite. The underlying genetics of refractoriness are complex and multifactorial. Completion of the genome sequence of An. gambiae 2 years ago, together with the development of DNA microarrays in this species and the extension of the RNAi technique to adult mosquitoes, has allowed comparative and functional genomic approaches of the mosquito innate immune system. A variety of factors were shown to negatively affect the development of Plasmodium parasites in the mosquito, in some cases leading to complete transmission blockage. In addition, mosquito factors have been identified that play positive roles and are required for successful transmission of the parasite. These findings indicate a highly complex interplay between parasite and vector. Research is continuing to identify new factors involved in this interaction and to decipher the interplay of these molecules and their regulation. PMID- 15894186 TI - Interactions among multiple genomes: tsetse, its symbionts and trypanosomes. AB - Insect-borne diseases exact a high public health burden and have a devastating impact on livestock and agriculture. To date, control has proved to be exceedingly difficult. One such disease that has plagued sub-Saharan Africa is caused by the protozoan African trypanosomes (Trypanosoma species) and transmitted by tsetse flies (Diptera: Glossinidae). This presentation describes the biology of the tsetse fly and its interactions with trypanosomes as well as its symbionts. Tsetse can harbor up to three distinct microbial symbionts, including two enterics (Wigglesworthia glossinidia and Sodalis glossinidius) as well as facultative Wolbachia infections, which influence host physiology. Recent investigations into the genome of the obligate symbiont Wigglesworthia have revealed characteristics indicative of its long co-evolutionary history with the tsetse host species. Comparative analysis of the commensal-like Sodalis with free living enterics provides examples of adaptations to the host environment (physiology and ecology), reflecting genomic tailoring events during the process of transitioning into a symbiotic lifestyle. From an applied perspective, the extensive knowledge accumulated on the genomic and developmental biology of the symbionts coupled with our ability to both express foreign genes in these microbes in vitro and repopulate tsetse midguts with these engineered microbes now provides a means to interfere with the host physiological traits which contribute to vector competence promising a novel tool for disease management. PMID- 15894187 TI - Using bacteria to express and display anti-parasite molecules in mosquitoes: current and future strategies. AB - Vector-borne diseases impose enormous health and economical burdens throughout the world. Unfortunately, as insecticide and drug resistance spread, these burdens will increase unless new control measures are developed. Genetically modifying vectors to be incapable of transmitting parasites is one possible control strategy and much progress has been made towards this goal. Numerous effector molecules have been identified that interfere with parasite development in its insect vectors, and techniques for transforming the vectors with genes encoding these molecules have been established. While the ability to generate refractory vectors is close at hand, a mechanism for replacing a wild vector population with a refractory one remains elusive. This review examines the feasibility of using bacteria to deliver the anti-parasitic effector molecules to wild vector populations. The first half briefly examines paratransgenic approaches currently being tested in both the triatomine bug and tsetse fly. The second half explores the possibility of using midgut bacteria to control malaria transmission by Anopheles mosquitoes. PMID- 15894188 TI - Global gene expression analysis of Anopheles gambiae responses to microbial challenge. AB - Anopheles gambiae transcript responses to experimental challenge with heat inactivated Salmonella typhimurium, Staphylococcus aureus and Beauveria bassiana have been analyzed with an Affymetrix GeneChip comprising the entire predicted mosquito transcriptome. Significant up- or down-regulation (greater than 2-fold) can be assayed for approximately 2% of the mosquito transcriptome and affected genes represent a variety of functional classes that include immunity, apoptosis, stress response, detoxification, metabolism, blood digestion, olfaction and others. Transcript responses to the 3 microbial elicitors exhibit an exceptionally high degree of specificity and only a few genes are significantly regulated by more than 1 of the tested elicitors. This study identifies several transcripts that have not been linked directly to immune response in A. gambiae previously; their infection responsiveness and sequence features do however suggest implication in defence reactions; examples are genes encoding leucine rich repeat domain proteins, cuticle domain proteins and proteins containing immunoglobulin and fibronectin domains. PMID- 15894189 TI - Ookinete-induced midgut peroxidases detonate the time bomb in anopheline mosquitoes. AB - Previous analysis of the temporal-spatial relationship between ookinete migration and the cellular localization of genes mediating midgut immune defense responses suggested that, in order to survive, parasites must complete invasion before toxic chemicals ("a bomb") are generated by the invaded cell. Recent studies indicate that ookinete invasion induces tyrosine nitration as a two-step reaction, in which NOS induction is followed by a localized increase in peroxidase activity. Peroxidases utilize nitrite and hydrogen peroxide as substrates, and detonate the time bomb by generating reactive nitrogen intermediates, such as nitrogen dioxide, which mediate nitration. There is evidence that peroxidases also mediate antimicrobial responses to bacteria, fungi and parasites in a broad range of biological systems including humans and plants. Defense reactions that generate toxic chemicals are also potentially harmful to the host mounting the response and often results in apoptosis. The two-step nitration pathway is probably an ancient response, as it has also been described in vertebrate leukocytes and probably evolved as a mechanism to circumscribe the toxic products generated during defense responses involving protein nitration. PMID- 15894190 TI - Is the mature endotoxin Cry1Ac from Bacillus thuringiensis inactivated by a coagulation reaction in the gut lumen of resistant Helicoverpa armigera larvae? AB - Bacillus thuringiensis endotoxins (Bt-toxins) are the most important biopesticides used in controlling insect pests and vectors of diseases. The emergence of widespread resistance to Bt in some insect species is a serious threat to agricultural production. Analysis of Bt-resistant and susceptible laboratory strains of Helicoverpa armigera revealed elevated immune responses involving increased melanization and the presence of a soluble toxin-binding glycoprotein in the hemolymph and gut lumen of the resistant strain. We propose a resistance mechanism against toxins based on a systemic immune-induction that can be transmitted to the next generation by a maternal effect. PMID- 15894191 TI - Insect renal tubules constitute a cell-autonomous immune system that protects the organism against bacterial infection. AB - Innate immunity is a widespread and important defence against microbial attack, which in insects is thought to originate mainly in the fat body. Here we demonstrate that the fluid-transporting Malpighian (renal) tubule of Drosophila melanogaster constitutes an autonomous immune-sensing tissue utilising the nitric oxide (NO) signalling pathway. Reverse transcriptase PCR (RT-PCR) shows that tubules express those genes encoding components of the Imd pathway. Furthermore, isolated tubules bind and respond to lipopolysaccharide (LPS), by upregulating anti-microbial peptide (diptericin) gene expression and increased bacterial killing. Excised, LPS-challenged tubules, as well as tubules from LPS-infected flies, display increased NO synthase (NOS) activity upon immune challenge. Targetted expression of a Drosophila NOS (dNOS) transgene to only principal cells of the tubule main segment using the GAL4/UAS system increases diptericin expression. In live flies, such targetted over-expression of dNOS to tubule principal cells confers increased survival of the whole animal upon E. coli challenge. Thus, we describe a novel role of Malpighian tubules in immune sensing and insect survival. PMID- 15894192 TI - On the distribution and genetic differentiation of Anopheles gambiae s.s. molecular forms. AB - This paper summarises published and unpublished data on the spatial and temporal distribution, and on the genetic characterisation of molecular forms M and S of Anopheles gambiae s.s. The two forms are characterised by a high level of gene flow restriction, by a largely overlapping geographical and temporal distribution, and by a low degree of genetic differentiation. Floating paracentric inversions on chromosome-2 are shown to be shared by the two forms, although with very different frequencies of alternative arrangements, confirming that these inversions are most probably involved in ecotypic adaptation, rather than in the building of reproductive barriers. Further studies and tools are needed to throw light on the genetic and biological differentiation of M and S to improve the knowledge of the real composition of the vector system, of its demography, population genetics and dynamics, also in view of the possible consequences on the transmission of human pathogens in sub-Saharan Africa. Preliminary results and perspectives of the use of transposable element insertion sites as markers of genetic differentiation and tools for population genetic studies are discussed. PMID- 15894193 TI - Association mapping of segregating sites in the early trypsin gene and susceptibility to dengue-2 virus in the mosquito Aedes aegypti. AB - Evidence suggests that midgut trypsins in Aedes aegypti condition the mosquito's ability to become infected with the dengue-2 flavivirus (DEN2). The activity of early trypsin protein peaks approximately 3 h after blood feeding and then drops within a few hours. We use association mapping to test the hypothesis that segregating sites in early trypsin condition midgut susceptibility to DEN2 virus. A total of 1642 females from throughout Mexico and the southern US were fed an artificial blood meal containing DEN2. After 2 weeks, mosquito heads and midguts were tested for DEN2. Mosquitoes with an infected head were classified as susceptible, those without a midgut infection had an infection barrier, and those with an infected gut but no head infection had an escape barrier. The early trypsin gene was amplified in two overlapping pieces from each mosquito and analyzed for single strand conformation polymorphisms (SSCPs). Unique SSCP genotypes were sequenced and 90 segregating sites were found. The dataset was divided into the four geographic regions within which Ae. aegypti is panmictic in Mexico. Heterogeneity chi2 analyses between alleles or genotypes and infection phenotypes demonstrated significant associations but allelic and genotypic effects were inconsistent among geographic regions. No consistent associations were found between segregating sites in early trypsin and susceptibility to DEN2 in Ae. aegypti in Mexico. PMID- 15894194 TI - Nanos (nos) genes of the vector mosquitoes, Anopheles gambiae, Anopheles stephensi and Aedes aegypti. AB - A number of genetics-based strategies for the control of vector-borne diseases require the development of genetic drive systems for introgressing antipathogen effector genes into wild populations of insects. Modified transposons whose mobilization is controlled by the DNA elements of developmentally regulated genes offer a potential solution for introducing effector genes into mosquitoes. Such elements could exhibit sex-, stage- and species-specific transposition, thus mitigating some of the concerns associated with autonomous transposition. Hybridizations in situ show that the transcription products of the nanos orthologous genes of Anopheles gambiae (Anga nos), An. stephensi (Anst nos) and Aedes aegypti (Aeae nos) accumulate in developing oocytes in adult females and localize to the posterior pole in early embryos. These features make nos genes promising candidates for donating control sequences to modified transposons. PMID- 15894195 TI - Economic burden of osteoporosis, breast cancer, and cardiovascular disease among postmenopausal women in an employed population. AB - BACKGROUND: Postmenopausal women have a significant risk of developing a number of chronic conditions including osteoporosis (OP), breast cancer (BrCa), and cardiovascular disease (CVD). These diseases can result in significant direct (medical treatment) and indirect (workplace) costs. The objective of this study is to assess these costs among an employed population. METHODS: Deidentified medical and disability claims data from seven large employers (n = 585,441) were analyzed from 1998 through 2000 for female employees, age 50-64 years. Medical claim ICD-9CM codes were used to identify patients treated for: OP (n = 2,314), BrCa (n = 555), and CVD (n = 1,710). Each disease cohort was compared to a random sample of 50- to 64-year-old female employees (n = 7,575). Descriptive and multivariate techniques were used to characterize direct and indirect costs attributable to each condition. RESULTS: Average annual direct costs were higher (p < .001) for female employees treated for OP (6,259 dollars), BrCa (13,925 dollars), or CVD (12,055 dollars) when compared with the random sample (2,951 dollars). In addition, average annual indirect costs associated with OP (4,039 dollars), BrCa (8,236 dollars), and CVD (4,990 dollars) were higher (p < .001) than indirect costs for the random sample (2,292 dollars). Even when controlling for each disease-state cohort's demographics and disease-specific comorbidities, patients treated for OP, BrCa, and CVD continued to have significantly greater direct and indirect costs (p < .001) than the random sample. CONCLUSIONS: Chronic conditions such as OP, BrCa, and CVD, which occur more frequently in women after menopause, impose a significant financial burden. Greater health care utilization and work-loss prevalence among women treated for these conditions contribute to these additional costs. PMID- 15894196 TI - Doulas as childbirth paraprofessionals: results from a national survey. AB - Fourteen randomized trials have demonstrated that continuous caregiver support during childbirth can lead to shorter labors and decrease the need for intervention. In response, there has been a significant increase in the number and use of doulas as paraprofessionals who provide social and emotional support to women during labor/birth for a fee. We conducted a mailed survey of a nationally representative sample of certified and certification-in-process doulas in the United States (n = 626, 64.4% response rate) to gather some descriptive information on their sociodemographic backgrounds, practice characteristics, and beliefs/attitudes on a number of salient issues. The survey results suggest that, in 2003, doulas were primarily white, well-educated married women with children. The majority of certified doulas worked in solo practice and provided childbirth support services on average to nine clients per year. Very few doulas were earning more than 5,000 dollars per year from this work, and only 10% of certified doulas reported receiving third-party reimbursement for their services. Thus, while almost all doulas found their work emotionally satisfying, only one in three saw their work as financially rewarding. Doulas also reported challenges in getting support/respect from clinicians and in balancing doula work and family life. In addition, one in four doulas reported that they were preparing for a career in midwifery. Doulas can play an important and unique role in the childbirth process and reap many personal rewards engaging in this type of work. However, a number of financial, personal, and professional challenges present significant obstacles to the growth of doulas as childbirth paraprofessionals in the United States. PMID- 15894197 TI - Threats to family planning services in Michigan: organizational responses to economic and political challenges. AB - Title X is the only federal funding specifically for contraception and family planning services. This study identifies the threats and challenges Title X family planning organizations face in Michigan, and examines organizational responses to these challenges. We hypothesized that organizational responses to current challenges, including recent legislation, would differ between organizational types. We used a multiple case study design to examine safety net providers that received Title X funding in 2001. Cases were selected to represent economic and geographic diversity and included a mix of population densities. Key informants at each organization participated in face-to-face, semistructured interviews. Interviews collected data on current challenges, organizational planning processes, and organizational responses. All Title X organizations reported significant challenges, including rising costs, increasing need, and inadequate funding. Private organizations were more concerned about political challenges, especially recent Michigan legislation, than health departments. Organizational type was associated with the type of response. Health departments tended to close clinics or cut services, whereas private organizations recruited insured populations and increased patient fees. Based on these findings, the family planning safety net in Michigan appears to be undergoing significant change. These changes may decrease the availability of affordable family planning services in Michigan. PMID- 15894198 TI - Gender differences in smoking cessation services received among veterans. AB - INTRODUCTION: Smoking is the leading preventable cause of death among women in the United States. It is a particular problem for women using the Veterans Health Administration (VA), where the prevalence of smoking among women is 30%. We compared the baseline characteristics of male and female smokers and then assessed the smoking cessation services they received to determine whether there are important gender differences in care. METHODS: As part of a study of implementing national guidelines for smoking cessation taking place at 18/23 VA centers in the southwestern and western United States, we conducted a baseline survey of a random sample of 1,941 smokers in primary care (129 women, 1,812 men) to assess the smoking cessation services received by female and male veterans. Subjects were followed 1 year later (73 women, 1007 men). Results for men and women were compared using chi-square tests and analysis of variance. Logistic regression analyses were conducted to determine factors that were independently associated with receipt of smoking cessation services. RESULTS: Female smokers were younger, more educated, and less likely to be married than male smokers. Women were equally likely to report being advised to quit smoking or referred to a smoking cessation program but were much less likely to report receiving a prescription for nicotine patches (OR .5, 95% CI .3-.9). One year later, female smokers were less likely to have successfully quit smoking. CONCLUSION: Women were less likely to report receiving nicotine patches for smoking cessation. Future interventions to increase use of smoking cessation medications for female smokers will also hopefully increase their quit rate. PMID- 15894199 TI - Portuguese-speaking women voice their opinions: using their words to teach about wife abuse. AB - BACKGROUND: This study examined Portuguese-speaking women's definitions of wife abuse and beliefs about appropriate responses to abuse. The goals were to determine the breadth of definitions and to examine cultural stereotyping. This information was sought in an effort to design education strategies in the Portuguese-speaking community. METHODS: One-on-one semistructured interviews were conducted with 163 Portuguese-speaking women living in Toronto, Canada. The participants' open-ended answers were first coded using Qualitative Software Research NUDIST and then narrowed and grouped. Responses were then tagged by group label to generate descriptive statistics using Statistical Package for Social Sciences. RESULTS: Participants defined wife abuse broadly with respect to kind, prevalence, and severity. Six themes for abuse emerged: physical, psychological, sexual, financial, patriarchal, and infidelity. Participants provided various responses to what women should do and actually do in response to abuse. Most participants believed that Portuguese-speaking women should leave their husbands and/or seek help for the abuse, but that they actually remain quiet about the abuse and take very little action. Their responses and explanations demonstrate that they hold stereotypes about their culture. CONCLUSIONS: The range of definitions reported by participants suggests that a continuum of wife abuse, using Portuguese-speaking women's own words, can be created to educate them about how abusive acts are linked and used to maintain control over women. The differences that emerged with respect to appropriate responses to abuse suggest that it may be important to challenge Portuguese speaking women about the cultural stereotypes that they hold to help them see patriarchy as widespread rather than particular to their culture. PMID- 15894200 TI - Body mass, depressive symptoms and menopausal status: an examination of the "Jolly Fat" hypothesis. AB - PURPOSE: Estrogens are linked with depression due to their ability to alter the function of the serotonin neural systems. We hypothesize that postmenopausal women should have a higher degree of depressive symptoms than premenopausal women. Further, because estrogen levels in postmenopausal women positively correlate with body fat, we hypothesize that there is an inverse relationship between body fat and depressive symptoms among postmenopausal women. METHODS: We enrolled 1156 Polish urban women aged 45 to 64 in a cross-sectional study. Depressive symptoms were assessed by the Center for Epidemiologic Studies Depression Scale (CES-D) scale. Menopausal status and education level was assessed by a standardized questionnaire. MAIN FINDINGS: Postmenopausal women had higher mean CES-D scores of depressive symptoms than premenopausal women (14.4 versus 13.2 respectively, p = .018). Both among pre- and postmenopausal women, those with higher education had lower scores of depressive symptoms. In addition, in postmenopausal women with lower education an inverse relationship was observed between body mass index (BMI) and depressive symptoms: a higher BMI was associated with a lower score of depressive symptoms (p = .009). Such a relationship was not present among premenopausal women or women who were postmenopausal but better educated. CONCLUSIONS: This study indicates that menopausal status is related to differences in the degree of occurrence of depressive symptoms. Our results support the "Jolly Fat" hypothesis for postmenopausal women with lower education, namely, a higher BMI is associated with lower score of depressive symptoms. PMID- 15894201 TI - [Evidence-based medicine: a new concept]. PMID- 15894202 TI - ["Don't mess with my gynaecologist!"]. PMID- 15894203 TI - [Clinical expression of precocious pubertal development in girls]. AB - The paediatric endocrinologist is frequently asked whether pubertal development in a girl is normal, early or too early (precocious). This review will cover all clinical expression of premature development of puberty: central precocious puberty (neurogenic, secondary, and idiopathic) where treatment with GnRHa is considered, early puberty, partial puberty or pubertal variants and peripheral or pseudo precocious puberty related to an antonomous hypersecretion of estrogens by the ovaries. A special attention should be paid also to the role of environmental disruptors in the development of peripheral precocious puberty. GnRHa treatment should be considered only when evidence of central activation of the gonadotropic axis is proved by the LHRH-test. PMID- 15894204 TI - [Thierry's spatula instrumental extraction: a study of foetal-maternal morbidity]. AB - OBJECTIVE: Assisted delivery is necessary in many obstetrical conditions but is involved in maternal and foetal complications. The legal pressure and the commendable aim consisting in less neonatal morbidity and mortality have called forth a reflection about the type and the way of instrumental foetal extraction. In 1950, Thierry had already felt this problem and he invented spatula to replace obstetrical forceps. Although this instrument appears empirically little deleterious, literature about its evaluation is very poor. We studied this instrument in a retrospective 190 cases series. PATIENTS AND METHOD: Retrospective study of 190 Thierry's spatula extractions, over a seven-year period (January 1996 to December 2002), at the Centre Hospitalier General of Montbeliard. RESULTS: Out of a total of 8126 deliveries for the study period, the instrumental extraction rate was 5.3%, with 40.6% spatula extractions (190 cases). No failure of Thierry's spatula extraction was noted. DISCUSSION ET CONCLUSION: Our study concludes that spatula is efficient but does not usually permit anterior rotation of occipito-posterior presentation. Maternal and foetal morbidity is not frequent. PMID- 15894205 TI - [Sentinel node biopsy without systematic axillary dissection: study about 1000 procedures]. AB - OBJECTIVE: To assess daily practice of 1000 sentinel node (SN) biopsies in breast cancer. PATIENTS AND METHOD: Prospective review of 1000 consecutive sentinel node biopsies between February 2001 and June 2004. Analyses concerned technical aspects of sentinel node detection, pathologic results of the tumor and sentinel node, treatment and follow-up. RESULTS: Nine hundred and seventy-eight SN were detected (98.7%). In univariate analyses, age, pathologic tumor size (20 mm) and method of detection (blue dye or isotopic vs. combined) were statistically significant. One hundred and fifty-six cases (16%) underwent immediate axillary dissection (AD), whereas 116 (12%) had a delayed AD. There were 923 invasive or micro-invasive carcinoma with detected SN: 282 SN (30.5%) were involved, either with macrometastases (166) or with micrometastases (116), 34% had positive non sentinel node. Age and metastasis size were predictive for AD involvement. Sixteen percent of micrometastatic SN had positive AD, there was no predictive factor for axillary involvement. After a median follow-up of 20 months, there were 4 axillary recurrences: 1 (0.1%) after negative SN without AD, 1 (0.1%) after positive SN with positive AD, 1 (4.3%) after micrometatastatic SN without AD, and 1 (8.3%) after macrometastatic SN without AD. There were 55 ductal carcinoma in situ and 54 micro-invasive cancer: positive SN (with negative AD) were detected in only 2 cases (2.3%). There were initially 112 ductal carcinoma in situ diagnosed by percutaneaous biopsy, 25 of them (22%) had invasive disease on definitive histology. Among there, 12 had involved SN (with 4 positive AD). DISCUSSION AND CONCLUSION: With a high detection rate and low recurrence rate, SN biopsy is considered in our institute as a reliable procedure and is used to evaluate regional nodal status of early breast cancer. Thus, 70% of AD can be omitted. PMID- 15894206 TI - [The efficacy and acceptability of mifepristone medical abortion with home administration misoprostol provided by private providers linked with the hospital: a prospective study of 433 patients]. AB - BACKGROUND: Until July 2001 medical abortion was only authorized in France in public hospitals or private clinics. A new law effective in July 2001 allows private practitioners (gynaecologists or general practitioners) to provide medical abortion in their offices as long as they are linked to a hospital official agreement. Unfortunately mifepristone was not available outside hospitals before September 2004, so the study was conducted still providing the drugs in the hospital family planning clinic. OBJECTIVE: To evaluate the efficacy and the acceptability of mifepristone medical abortion with home administration of misoprostol provided by private practitioners linked with the hospital. PATIENTS AND METHODS: Four hundred thirty-three women seeking medical abortion before 7 weeks LMP were included between 2 January 2003 and 7 July 2004. All consultations before abortion and 2 weeks after abortion took place in a private provider's office. Drugs were administrated in the hospital family planning clinic: patients were given 3 tablets of mifepristone (600 mg) orally by the midwife and received 2 tablets of misoprostol (400 microg) that they would take at home 48 hours later. In case of any problems or complications, patients could phone or meet their private providers, phone the hospital midwife or go to the hospital emergency service. Private providers received training in medical abortion training and could at any time reach a medical specialist in the hospital family planning clinic for information or to refer a patient. RESULTS: - Efficacy was evaluated for 339 women, because 94 patients were lost to follow-up (21.7%). Efficacy of medical abortion was 93.8% (318/339). There were 21 surgical aspirations (6.2%): for women's decision in 1.5% of cases, for medical decision without complications in 3.5% of cases, and for failure of the method in 1.2% of cases (2 ongoing pregnancies and 2 heavy haemorrhages with transfusion). The family planning midwife received a phone call from 21 patients after mifepristone (4.8%), Twenty-five patients had an emergency consultation (5.7%), and 22 patients went back to their private providers before their appointment for follow up (5%). Twenty-two patients (5%) were referred by the private provider to the hospital medical specialist. Acceptability is known for 26% of patients; 96.2% thought that the abortion procedure was acceptable. DISCUSSION AND CONCLUSIONS: The failure rate of medical abortion in this study is largely due to aspirations for incomplete abortion. To improve the efficacy of medical abortion offered by private providers linked with the hospital, all the relevant professionals (private providers, residents in the emergency service, family planning providers) must be well trained in medical abortion, especially in how to interpret and react to ultrasound images obtained in the follow up visit. The procedure is very acceptable to women. Medical abortion offered via a network should be well accepted by practitioners, since only 5% of women will need more than two consultations and only 6.2% will need surgical aspiration in the hospital. This study allows us to be optimistic about the expansion of medical abortion in France outside the hospital via a provider-hospital network based on the fact that since September 2004 private providers can get mifepristone directly in the pharmacies of the city. PMID- 15894207 TI - [Prenatal ultrasound diagnosis of neuroblastoma]. AB - With the progress of foetal ultrasonography, diagnosis of foetal tumors can be made before birth. We report five cases of prenatally detected neuroblastoma of the adrenal glands. Diagnosis was made during the third trimester in all cases. At birth no clinical sign related to the tumor was present, and urinary catecholamines were within normal ranges. On ultrasound scans the tumor was cystic in 1 case, solid in 2 cases and of mixed echogenicity in 2 cases. All children were operated on without pre or postoperative chemotherapy. In each setting the tumor was a stage I neuroblastoma according to the Evans classification. All children are alive and disease-free with a follow-up of 32 months at 14 years. PMID- 15894208 TI - [Laparoscopic total hysterectomy after radiochemotherapy in an obese woman with neuroendocrine carcinoma of the cervix: surgical and anesthesiological aspects]. AB - Massive obesity is an important risk factor in gynaecologic surgery. The traumatic effect of traditional laparotomy on the parietal wall is responsible for important perioperative morbidity. We describe the first reported case of an obese woman (Body Mass Index = 55 kg/m2) with stage IIA neuroendocrine carcinoma of the cervix treated by laparoscopy after radiochemotherapy. After a complete response to radiochemotherapy, the patient underwent laparoscopic hysterectomy and bilateral salpingo-oophorectomy. The laparoscopic procedure was performed with a low-pressure pneumoperitoneum. She was discharged at day 2. No major complication was observed. Surgical and anesthesiological laparoscopic management in obese women are discussed. PMID- 15894209 TI - [Pre-implantation genetic diagnosis and spontaneous pregnancies: an unexpected acting out]. AB - OBJECTIVES: To understand the psychological motives that led these couples to take risks involved in genetic transmission and the possibility of another spontaneous pregnancy. To report our psychoanalytic experience to explain why after a request of pre-implantation genetic diagnosis (PGD), some women prefer to be spontaneously pregnant. PATIENTS AND METHOD: Between 2000 and 2004, out of 643 requests of PGD, 541 have been admitted, among which 123 spontaneous pregnancies appeared during the process of PGD. RESULTS: Some encounters with the patients have shown some reasons to these spontaneous pregnancies - denial of IVF, ambivalence of this desire for a child, denial of the risk, control of the procreation, family loyalty and fertility affirmation. DISCUSSION AND CONCLUSION: The resort to PGD is more ambivalent than the thought of it because it means, beyond the request, conflict with transmission, culture, and history origins. PMID- 15894210 TI - [Women with endometriosis: are they different from others?]. AB - The objective of this short review is to identify the particularities of women with endometriosis, especially those complaining of pain and with the most severe lesions. Genetic aberrations play, with a high probability, a major role in the development of this disease, its severity, its tendency to recur and also in its capacity to degenerate. The abnormalities of the endometrium, with exacerbated biological activities, are an example. The woman with endometriosis seems more sensitive to pain through various mechanisms, such as central hypersensitivity and decrease threshold to somatoceptive pain and several associated psychological disorders. Endometriosis is often associated with other painful conditions such as irritable bowel syndrome, interstitial cystitis and fibromyalgia. Finally, also appears also to have a higher risk to develop non Hodgkin's lymphoma or ovarian cancer. These particularities, some of them being still speculative or controversial, should be known in routine practise, in order to offer a better multidisciplinary management, not only for short term, but also long term issues. PMID- 15894211 TI - [Diagnostic hysteroscopy and risk of peritoneal dissemination of tumor cells]. AB - Questions have been raised about the safety of diagnostic hysteroscopy preceding surgical treatment of endometrial carcinoma. Several studies showed that the risk of a positive cytology among patients presenting endometrial adenocarcinoma was increased after diagnostic hysteroscopy, suggesting a peritoneal dissemination of tumor cells due to the exploration. We studied this hypothesis on the basis of a systematic review of the scientific data. Five studies fulfilling inclusion criteria have been selected and have been introduced into a fixed model of meta analysis. On a total of 756 studied patients, 79 presented a positive peritoneal cytology. The diagnostic hysteroscopy did not increase significantly the risk of abdominal dissemination of tumor cells, the peritoneal cytology being positive among 38 patients in the group having undergone this intervention vs 41 patients in the control group (OR = 1,64; 95% CI: 0,96-2,80). In conclusion, no formal evidence is currently available concerning the role of diagnostic hysteroscopy on the frequency of peritoneal dissemination of tumor cells, or on the vital prognosis of the patients presenting with endometrial carcinoma. From the data available, there is not any reason to avoid diagnostic hysteroscopy in the initial workup of endometrial cancer. PMID- 15894212 TI - [Zinc and pregnancy]. AB - Zinc, an essential trace element, plays a critical role in normal growth and development, cellular integrity and many biological functions, including protein synthesis and nucleic acid metabolism. Since all these are involved in cell division and growth, zinc is believed to be important for foetal growth and development. Zinc requirement is increased during pregnancy but the lack of a valid indicator precludes a true estimate of zinc deficiency in pregnancy even in developed countries. This review examines the possibility that a gestational deficiency of zinc can adversely affect the pregnancy outcome. Preliminary human data suggest a beneficial effect of prenatal zinc supplementation trials in particular on infant's neurobehavioral development. In the light of the currently available information, zinc supplementation at therapeutic load (30 mg/day) as it is proposed for the treatment of hormonal skin disorders to adolescents, cannot be toxic. PMID- 15894213 TI - [Licensed or non-licensed tocolysis?]. PMID- 15894214 TI - [Tocolysis by first intention with atosiban]. PMID- 15894215 TI - [Tocolysis by first intention with nifedipine]. PMID- 15894216 TI - [Postpartum haemorrhage: methods used and topics discussed in the CNGOF Recommendations for clinical practice (December 2004)]. PMID- 15894217 TI - [Postpartum haemorrhage: recommendations for clinical practice by the CNGOF (December 2004)]. PMID- 15894218 TI - [Response of Jeanine Henry-Suchet to the article by F. Hamdad et al]. PMID- 15894219 TI - [Response of P. Judlin and O. Thiebaugeorges to the article by F. Hamdad et al]. PMID- 15894221 TI - Proton pump inhibitors to be used more? PMID- 15894222 TI - Genotypic guidance for chemotherapy choices. PMID- 15894223 TI - Mark Fishman on changing the grammar of drug discovery. Interviewed by Christopher Watson. PMID- 15894224 TI - When counting sheep does not help: the growing incidence of sleep disorders. PMID- 15894225 TI - The promise of biomarkers: research and applications. PMID- 15894226 TI - Peroxidases: a role in the metabolism and side effects of drugs. AB - Current safety screening of drug candidates or new chemical entities for reactive metabolite formation focuses on the role of cytochrome P450. However, peroxidases also have a major role in drug metabolism, and peroxidase-catalyzed drug oxidation could lead to reactive metabolite formation, resulting in oxidative stress and cytotoxicity. Here, the different classes of human peroxidases are summarized and the molecular mechanisms of peroxidase-catalyzed drug metabolism are discussed. In addition, evidence is presented that indicates a role of these enzymes in drug toxicity. PMID- 15894227 TI - cGMP-dependent protein kinases in drug discovery. AB - Cyclic guanosine-3', 5'-monophosphate (cGMP)-dependent protein kinases (cGKs) are key enzymes of nitric oxide-cGMP and natriuretic peptide signalling cascades. These kinases mediate most of the effects of cGMP-elevating drugs, such as nitrates and phosphodiesterase inhibitors. cGKs modulate smooth muscle relaxation (e.g. the vasculature, gastrointestinal tract, bladder and penile), platelet aggregation, renin release, intestinal secretion, learning and memory. Furthermore, several cGK substrates have been identified. Isozyme-specific inhibitors and activators of cGK and its downstream substrates might act more specifically than upstream signalling activators, such as organic nitrates and phosphodiesterase inhibitors. PMID- 15894228 TI - FT-ICR mass spectrometry in the drug discovery process. AB - The high mass accuracy and resolution of Fourier transform (FT)-ion cyclotron resonance (ICR) mass spectrometry are making it an increasingly useful tool in drug discovery and development. The basics of FT-ICR are described here, including modern ion sources and fragmentation methods. Although FT-ICR is not a high-throughput method in the traditional sense, previously difficult and complex problems are being efficiently approached using steadily improving instruments and magnets. Applications are surveyed in fields such as proteomics, metabonomics, natural product analysis and non-covalent complexes. PMID- 15894229 TI - Staphylococcus aureus: the search for novel targets. AB - In the UK, 20,000 cases of Staphylococcus aureus bacteraemia are reported each year, half of which are antibiotic resistant and approximately 4% are fatal, exemplifying a worldwide phenomenon of tremendous economic and human impact. Novel treatments and prophylaxis are urgently required to combat such a serious threat. A common goal in the postgenomic era is to identify new targets for drug intervention (using small molecules) and immunologicals. Several promising cellular targets are now being developed in the quest to control such a life threatening pathogen. PMID- 15894230 TI - Biological networks and analysis of experimental data in drug discovery. AB - Cellular life can be represented and studied as the 'interactome'--a dynamic network of biochemical reactions and signaling interactions between active proteins. Systemic networks analysis can be used for the integration and functional interpretation of high-throughput experimental data, which are abundant in drug discovery but currently poorly utilized. The composition and topology of complex networks are closely associated with vital cellular functions, which have important implications for life science research. Here we outline recent advances in the field, available tools and applications of network analysis in drug discovery. PMID- 15894231 TI - In silico strategies for modeling membrane transporter function. AB - Transporter proteins facilitate the transfer of solutes across the cell membrane and have an intricate role in drug absorption, distribution and excretion. Because of their substrate promiscuity, several transporters represent viable pharmacological targets for enhancing drug absorption, preventing drug toxicity or facilitating localized tissue delivery. However, the slow emergence of high resolution structures for these proteins has hampered the intelligent design of transporter substrates. Nonetheless, currently available functional, as well as structural, data provide an attractive scaffold for generating fusion models that merge substrate-based SARs and protein-based homology structures. The resultant models offer features that extend single modality paradigms in predictive function. PMID- 15894234 TI - Can we justify the cost of echocardiography? Lessons from outcomes research. AB - The inexorable growth in the performance of cardiac imaging studies has led to a significant reduction in reimbursement for these services, which are already being reimbursed at levels that are quite similar to their cost. This paper seeks to apply the principles of analysis of cost-effectiveness to echocardiography. Opportunities are summarized for reducing cost and maximizing effectiveness, and different approaches to defining cost-effectiveness are examined for valvular, myocardial and ischemic indications. PMID- 15894235 TI - Usefulness of quantitative intravenous myocardial contrast echocardiography to analyze microvasculature perfusion in patients with a recent myocardial infarction and an open infarct-related artery: comparison with intracoronary myocardial contrast echocardiography. AB - AIMS: We analyzed the usefulness of quantitative intravenous myocardial contrast echocardiography to study microvasculature perfusion after infarction in comparison with intracoronary myocardial contrast echocardiography. METHODS AND RESULTS: Thirty-two patients with a first ST elevation myocardial infarction, single-vessel disease and an open artery (TIMI 3) were studied before discharge. Myocardial perfusion in the risk area was quantified with intracoronary and intravenous myocardial contrast echocardiography. Perfusion was normal (intracoronary contrast echocardiography normalized videointensity >0.75) in 78 out of 97 dysfunctional segments (80%). Sensitivity and specificity of intravenous contrast echocardiography to predict normal perfusion were 87% and 63% for 'first-pass myocardial blood flow' (upslope of contrast arrival x peak intensity after intravenous bolus injection of contrast) and 91% and 89% for end systolic single-triggered images captured every 6 cycles, respectively. In an analysis per patients, normal perfusion (0 or 1 hypoperfused segments with intracoronary contrast echocardiography) was observed in 22 cases (69%). End systolic single-triggered images showed a strong correlation with intracoronary contrast echocardiography (R2 = 0.82, p = 0.0001). CONCLUSIONS: Intravenous contrast echocardiography is a useful technique to analyze microvasculature perfusion soon after infarction. A quantitative analysis of single-triggered images is an easy-to-obtain and reliable method to define perfusion when compared with intracoronary contrast echocardiography. PMID- 15894236 TI - Age-related intramyocardial patterns in healthy subjects evaluated with Doppler tissue imaging. AB - BACKGROUND: The aim of this study is to analyse spatial distribution of myocardial velocities (MV) and myocardial velocity gradient (MVG) with color M mode Doppler tissue imaging (DTI) and to analyse the influence of age in such parameters. METHODS AND RESULTS: A prospective study including 66 healthy volunteers was carried out with color M-mode DTI. Postprocessing of images was performed using proprietary software allowing the division of the myocardial wall into subendocardium, mesocardium and subepicardium. MV corresponding to the three layers and MVG time curves were obtained and systolic, early diastolic and late diastolic peak values were identified. MV were highest in subendocardium in systole, protodiastole and telediastole compared to external layers. Protodiastolic peak MV decreased in all layers with age, but with a higher impact in the subendocardium (r = 0.72, b = 0.136 (IC 95% 0.107-0.164), p = 0.0005). Older age resulted in larger telediastolic peak MV, without significant differences among layers. Linear correlation between protodiastolic peak mitral flow and peak protodiastolic velocity was higher in endocardium than in other layers (r = 0.79, p = 0.0005). CONCLUSIONS: Color M-mode DTI multilayer analysis showed that endocardium is more susceptible to age-related changes involving diastolic function. This dependency on age should be considered when assessing MV in other clinical settings. PMID- 15894237 TI - Ultrasonic strain/strain rate imaging--a new clinical tool to evaluate the transplanted heart. AB - OBJECTIVE: The aim of this study was to investigate the clinical applicability of strain and strain rate imaging (epsilon/SRI) in heart transplantation (Htx) patients and to establish "normal" post-Htx regional systolic deformation values. BACKGROUND: Epsilon/SR indices have been shown to be a more sensitive measure of regional systolic function than standard echo measurements. Thus, they might provide a new tool to better define both normal cardiac graft function and detect changes due to post-Htx complications. However, prior to investigating the role of epsilon/SRI in detecting abnormalities, "normal" post-Htx regional deformation values must be established as graft regional function can be altered by a number of factors such as ischemic time, surgical technique or accelerated graft ageing. METHODS: A total of 57 Htx patients (age 36+/-12 years; post-Htx 5.5+/-3 years) without any documented complication were studied. Epsilon/SRI data were acquired from the septum, left ventricular (LV) free walls and right ventricular free wall (RVFW). A total of 29 age-matched healthy subjects served as controls. RESULTS: Htx longitudinal peak systolic velocities (Vsys) were lower in inferior, septal and RVFW segments compared to controls. Peak systolic epsilon/SR (epsilonsys/SRsys)) did not differ from controls except in septum and RVFW in which the values were significantly reduced. Radial Vsys) in the Htx group were higher than controls while epsilonsys/SRsys were reduced. There was a significant decrease in SR(sys) in apical LV segments with increasing time post-Htx, whereas those measured in RVFW showed an increase by that time. CONCLUSION: Epsilon/SRI demonstrated that "healthy" Htx hearts have normal global systolic function but altered regional systolic deformation indices compared to normal hearts. Post-Htx time has a diminishing effect on the regional systolic deformation indices in LV segments but an improving effect in RVFW. These "normal" Htx values should provide the basis for subsequent studies into the role of epsilon/SRI in the non invasive detection of post-Htx complications. PMID- 15894238 TI - Impact of hand-carried cardiac ultrasound on diagnosis and management during cardiac consultation rounds. AB - AIM: To evaluate the impact of hand-carried cardiac ultrasound (HCU) on the diagnosis and management of patients during cardiac consultation rounds. METHODS AND RESULTS: One hundred and fifty patients hospitalized in non-cardiac units were included after the consulting cardiologist felt that an echocardiographic examination was indicated as part of his work-up. They were randomly allocated to echocardiography with an HCU device (SonoHeart, SonoSite, Inc.) (75 patients) or with a full-featured standard echo (FE) system (75 patients). The consulting cardiologist noted whether a definitive diagnosis was made or further study was necessary. Diagnosis and change in management were noted. In the HCU patient group there were 103 clinical questions. Seventy-two percent of the referral questions required no comprehensive echocardiographic evaluation. For questions of left ventricular function, valve abnormalities and pericardial effusion this was 98%. In 48% there was an immediate change in clinical management. In the FE patient group there were 94 clinical questions. In 32% the FE examination led to change in clinical management. CONCLUSION: HCU echocardiography provides clinically worthwhile assessment of left ventricular function, valve abnormalities and pericardial effusion in 98% of the cases. A direct assessment of cardiac function and anatomy at the bedside by an experienced cardiologist results in a significant immediate change in clinical management during consultation. PMID- 15894239 TI - Improvement in diastolic left ventricular function after coronary artery bypass grafting as assessed by recordings of mitral annular velocity using Doppler tissue imaging. AB - AIMS: The aim of the study was to evaluate the changes in diastolic function after coronary artery bypass grafting (CABG), using pulsed-wave Doppler tissue imaging (DTI). METHODS: Fifty-three patients with coronary artery disease were studied before and 3 and 12 months after CABG. Using pulsed-wave DTI, the mitral annular velocities were determined at 4 sites in the left ventricle (LV). Patients were also examined with dobutamine stress echocardiography and myocardial scintigraphy before and 3 months after CABG. RESULTS: The conventional transmitral velocity profiles were unchanged after CABG. DTI showed a marked improvement in diastolic LV function after CABG (early diastolic velocity: 7.5+/ 1.9, 8.2+/-1.7 and 9.3+/-2.7 cm/s before and 3 and 12 months after CABG, respectively, P < 0.01). The improvement in early diastolic velocity was more pronounced in patients showing no sign of residual ischemia in comparison to those with residual ischemia determined by myocardial scintigraphy (7.41+/-2.04 vs. 9.25+/-2.61 cm/s, P < 0.01 in the nonischemic group; 7.29+/-2.16 vs. 8.41+/ 2.55 cm/s, n.s., in the ischemic group). Before CABG, a significant increase in the systolic velocity (6.4+/-1.3 vs. 8.7+/-2.5 cm/s, P < 0.001), but not the early diastolic velocity (7.6+/-1.9 vs. 8.0+/-2.2 cm/s), was noted during stress echocardiography. Three months after CABG, both the systolic (6.5+/-1.3 vs. 9.3+/ 2.8 cm/s, P < 0.001) and the early diastolic velocities (8.1+/-1.8 vs. 10.3+/-2.2 cm/s, P < 0.001) improved during stress echocardiography. CONCLUSION: The results of the present study show that diastolic function improves at rest and under stress in patients after CABG. The improvement was seen only in patients without postoperative signs of reversible ischemia. PMID- 15894240 TI - Improved stroke volume assessment in the aortic and mitral valves with a new method in subjects without regurgitation. AB - BACKGROUND: Echocardiography combining Doppler and two-dimensional data is recommended for quantitative assessments of valvular regurgitation. We applied a new method to calculate the mitral annulus (MA) area in combination with multiple sample sites. Individuals without regurgitation in whom the valvular and left ventricular stroke volumes (SV) should be identical were investigated in order to evaluate the feasibility in quantitative assessments of valvular regurgitation. METHODS AND RESULTS: Twenty subjects were included. Flow velocity was registered with pulsed Doppler in different positions in the left ventricular outflow tract (LVOT) and in the MA. The MA area was assumed to be either circular, using the diameter from a four-chamber projection, or elliptic, using the major diameter from a parasternal short axis and a minor diameter from an apical long axis. Left ventricular (LV) SV was measured from LV volumes using the biplane method. The overall difference between LVOT SV and mitral SV using one centrally located measurement and elliptic MA was 3.2+/-15.6 ml (P=0.38), 0.9+/-15.7 ml between LVOT SV and LV SV (P=0.80) and -2.2+/-15.2 ml between mitral SV and LV SV (P=0.54). The corresponding standard deviation of the differences as a percentage of the mean value was 24%, 25% and 23%. A circular shaped MA overestimated the mitral SV compared with LVOT SV (P=0.009) and LV SV (P=0.004). Increasing the number of sample sites in the LVOT or MA did not further improve the results. CONCLUSION: Doppler and two-dimensional echocardiography can be used to quantify regurgitation in groups of patients. In individual patients the wide distribution of differences between valves and LV SV implies that the method should be used in conjunction with other Doppler echocardiographic parameters. PMID- 15894241 TI - Imaging of isolated ventricular non-compaction with dynamic cardiomyoplasty. PMID- 15894242 TI - Rare cause of right heart failure: contained rupture of a sinus of Valsalva aneurysm associated intraventricular septal aneurysm. AB - Sinus of Valsalva aneurysm (SVA) is a rare congenital lesion described first in 1840 by John Thurnam (Cited by Boutefou JM, Moret PR, Hahn C, Hanf E. Aneurysms of the sinus of Valsalva: report of seven cases and review of the literature. Am J Med 1978;65:18-24). In most cases unruptured Sinus of Valsalva aneurysm (SVA) is clinically silent; however, if it progressively enlarges it may cause coronary artery compression, complete heart block, or right ventricular outflow tract obstruction (Meier JH, Seward JD, Miller FA, Oh J, Sarano ME. Aneurysms in the left ventricular outflow tract: clinical presentation, causes, and echocardiographic features. J Am Soc Echocardiogr 1998;11:729-45; D'Silva SA, Dalve BV, Lokhandwala YY, Kale PA, Tendolkar AG. Unruptured congenital aneurysm of the left sinus of Valsalva presenting as acute right heart failure. Chest 1992;101:578-79) or is a potential source of cerebrovascular emboli. (Shahrabani RM, Jairaj PS. Unruptured aneurysm of the sinus of Valsalva: a potential source of cerebrovascular embolism. Br Heart J 1993;69:266-67). In this report, we describe a case of right coronary sinus of Valsalva aneurysm with a contained rupture. The containing rupture is in intraventricular septal aneurysm; the patient presents with right-sided heart failure. PMID- 15894243 TI - Primary left heart malignant fibrous histiocytoma. AB - We present the case of a 53 years-old woman presenting with congestive heart failure and pleural and pericardial effusions, in whom transthoracic and transesophageal echocardiography revealed multilocular cardiac tumor involving the left atrium wall, extending into the pericardium. Tumor was excised surgically and proved to be a malignant fibrous histiocytoma, primarily confined to the heart. Despite surgery followed by chemotherapy, the patient died a few months later. This is the 47th case of primary cardiac fibrous malignant histiocytoma reported in the literature. The pericardial involvement has been only rarely reported. Echocardiography was the method of diagnosis before rapid referral to surgery. PMID- 15894244 TI - Acute ischemia as a complication by transoesophageal echocardiography. AB - Transoesophageal echocardiography (TEE) is a safe procedure with a low complication rate. We present a patient with severe coronary artery stenosis or disease who developed acute coronary ischemia of the anterior wall as a complication of TEE procedure. The possible mechanism can be stress during the procedure. PMID- 15894245 TI - Multicavitated left atrial myxoma mimicking a hydatid cyst. AB - A 67 year-old asymptomatic patient was referred to our echo-lab because of hypertension. Transthoracic 2D-echocardiogram showed a non-prolapsing ovoid mass attached to the left side of the interatrial septum. Transesophageal echocardiography evidenced an ovoid cavitated mass with internal areas of calcification. Color Doppler revealed flow inside the cavities. At surgery, a multicavitated mass was observed attached to interatrial septum. Macroscopically revealed cavities filled with blood, as well as partially calcified areas. Microscopically there were collections of "lipidic" cells embedded in the myxoid matrix, typical of cardiac myxoma. Cardiac hydatid cysts usually have a rounded shape. Most myxomas are solid masses without a cystic architecture or cavitations. Calcification is usually identified at a microscopic level. The combination of a polycystic appearance of the mass and macroscopic areas of calcification is more frequently observed in hydatid cysts than in cardiac myxomas. This appearance of the mass leads us to consider a cardiac echinococcal cyst as the first diagnostic possibility. This peculiar structure of cardiac myxoma, to the best of our knowledge, has never been documented. Transthoracic echocardiography and particularly transesophageal imaging, enable us to delineate this kind of tumors. Surgical resection is the appropriate treatment for these tumors, even in asymptomatic patients. PMID- 15894246 TI - Improving the quality of residential care using goal attainment scaling. AB - OBJECTIVE: To illustrate the potential of goal attainment scaling (GAS) as a means to improve the quality of care provided by residential care facilities to frail older adults. DESIGN: A pre-test/post-test single-group design. SETTINGS/PARTICIPANTS: Twenty facilities providing inadequate care to at least one resident. These facilities were identified with a case-finding questionnaire followed by a thorough investigation of the quality of care delivered to a sample of frail older adults. INTERVENTION: The 6-month intervention was conducted by three interdisciplinary teams of health professionals experienced in caring for frail older adults. The intervention was tailored to the main quality problems identified at baseline in the facility. The first task of the intervention team was to set weighted priority goals in conjunction with the facility manager. Subsequent monthly on-site visits, interspersed with frequent telephone calls, were devoted to assisting the manager and staff implement permanent changes in the areas of care targeted for improvement. MEASUREMENTS: Pre- and post intervention GAS scores. RESULTS: Two facilities were lost to follow-up. A total of 81 facility goals were established at the onset of the study. Goals per facility ranged from 2 to 9 (mean = 4.5, SD = 2.04). GAS scores increased significantly between pre- and post-intervention (P < .001). Goal achievement varied with the manager's own rating of the quality of care provided in the facility (P = .008), his/her education level (P = .037), and the intervention team (P = .049). CONCLUSIONS: This study shows that quality improvement objectives established with the manager following a thorough evaluation of the quality of care provided in the facility can, on average, be attained with the help of experienced health professionals. It also provides some insight into the types of facilities where preset objectives are more likely to be achieved. PMID- 15894247 TI - Quality improvement in nursing homes in Texas: results from a pressure ulcer prevention project. AB - BACKGROUND: Pressure ulcer prevalence, cost, associated mortality, and potential for litigation are major clinical problems in nursing homes despite guidelines for prevention and treatment. OBJECTIVE: To improve the use of pressure ulcer prevention procedures at nursing homes in Texas through implementation of process of care system changes in collaboration with a state quality improvement organization (QIO). DESIGN: Preintervention and postintervention measurement of performance for process of care quality indicators and of pressure ulcer incidence rates. SETTING: Twenty nursing homes in Texas. PARTICIPANTS: Quality improvement teams at participating nursing homes. MEASUREMENT: Data were abstracted from medical records on performance measures (quality indicators) and pressure ulcer incidence rates between November 2000 and August 2002. Descriptive and inferential statistics were used. INTERVENTIONS: Process of care system changes consisting of tools and education to prevent pressure ulcers were introduced to participating nursing homes. RESULTS: Participating nursing homes showed statistically significant improvement in 8 out of 12 quality indicators. Pressure ulcer incidence rates also decreased, although not quite significantly. Furthermore, facilities with the greatest improvement in quality indicator scores had significantly lower pressure ulcer incidence rates than the facilities with the least improvement in quality indicator scores (S = 131.0, P = .03). This suggests that the interventions positively affected not only the process of care but also led to a decrease in pressure ulcer incidences. CONCLUSIONS: These results show that nursing homes in a collaborative effort with a QIO were able to improve their processes of care. Although significant improvement was noted on most of the quality indicators, opportunity remains for further improvement. Furthermore, these results suggest that implementation of process of care system changes by nursing homes in a collaborative relationship with a QIO may yield improvements in measures of patient outcome (eg, pressure ulcer incidence). PMID- 15894248 TI - Impact of the Eden Alternative on family satisfaction. AB - OBJECTIVES: The purpose of the study was to determine the impact of the Eden Alternative on family satisfaction. DESIGN: The study was a one-group pre-post design. Families were surveyed prior to the beginning of implementation of the Eden Alternative and again 2 years later. SETTING: The study was conducted at a 150-bed nursing home in the Midwest. PARTICIPANTS: Thirty-seven family members completed the survey at both time points. INTERVENTION: More than 80 in-services on the 10 principles of the Eden Alternative were provided to the staff on all shifts over the first year. Implementation of the Eden Alternative began during the second year. MEASUREMENTS: The Family Questionnaire, which measures perception of skill and caring nature of caregivers, quality of activities, the environment, contentment of elders, and relationships with administration, was mailed to a family member of each elder living in the nursing home in March of 2002 and again in March of 2004. RESULTS: There was a significant improvement in family satisfaction, as measured by the Family Questionnaire (P < .0001) after implementation of the Eden Alternative. CONCLUSION: The Eden Alternative provided many opportunities for family involvement. The improved satisfaction scores reflected greater communication and interaction among families, staff, and residents. PMID- 15894249 TI - Current status and future promise of pharmacotherapeutic strategies for Alzheimer's disease. AB - BACKGROUND AND OBJECTIVE: The pharmacotherapy of Alzheimer's disease (AD) is evolving rapidly. Unless new discoveries continue to emerge to facilitate prevention and effective treatment of the disease, the anticipated burden of this disease on caregivers and society at large will overwhelm resources. The objective of this paper is to review the state of development of approaches likely to yield effective interventional measures with regard to AD in the future. DESIGN: A comprehensive systematic search of MEDLINE using focused search criteria, a search of reference lists from these studies and reviews, a review of the Cochrane Database of Systematic Reviews, and a hand search of relevant journals was conducted. Selection of articles was based on the clinical focus. Additional inclusion criteria preferentially selected key articles that contained higher-level evidence in accordance with explicit, validated criteria. RESULTS: Pharmaceutical interventions are being developed and tested that confer neuroprotective benefits by targeting causative mechanisms. CONCLUSION: The paradigm that AD is pharmacologically unresponsive is shifting. Our understanding of the molecular mechanisms of neurodegeneration will soon allow us to more specifically target and interrupt the processes that contribute to this dementia. PMID- 15894250 TI - What is the proper use of hemoglobin A1c monitoring in the elderly? AB - Diabetes mellitus (DM) is a major health problem for the aging population. Glycemic control is fundamental to the management of diabetes, as glycemic levels are closely linked to development of diabetes-related complications. Measurement of the hemoglobin A1c (A1c) to assess chronic glycemic control is an integral component of diabetes care. Currently, there is no clear evidence that age alters the relationship between A1c and average blood glucose. The Diabetes Control and Complications trial and the United Kingdom Prospective Diabetes Study are the 2 main studies that have provided evidence leading to the widespread recommendation of A1c monitoring. The American Diabetes Association recommends achieving an A1c level of 7% or lower. However, older diabetics represent a heterogeneous population ranging from frail nursing home residents to active community-dwelling elderly with variable life expectancies. One needs to look at the individual in order to best balance risk versus benefit associated with tight glycemic control. Benefits of intensive therapy in an effort to lower A1c must always be weighed against the greater risk of disabling and unpredictable hypoglycemia, as the geriatric population is less likely to benefit from reducing the risk of microvascular complications and more likely to suffer serious adverse effects from hypoglycemia. PMID- 15894251 TI - Transition through the continuum of care in a continuing care retirement community: can a functional rating scale be a decision-making tool? AB - This retrospective descriptive study was done to determine if the Geriatric Functional Rating Scale (GFRS) can be used as an objective measure of a resident's need for a higher level of care in a continuing care retirement community (CCRC). A total of 290 of the 300 available independent living residents in a CCRC underwent assessment with the GFRS. At the end of the study period (February 2000 through December 2003), 84% of those who initially had a score of > or =40 were still living independently (alone or with a spouse), of those scoring 20 to <40, 11% were living independently, 40% required some assistance, and 20% entered a Long-Term Care Facility (LTC). Of those scoring <20 initially, none were living independently at the end of the study period. GFRS scores of > or =40 seemed to correlate with continued independence and <40 with an increasing need for assistance. The GFRS provides a functional database on each resident and may be a good objective predictor of future level of care. The sensitivity and specificity of GFRS scores <40 were 42% and 96%, respectively, for either admission to a facility or death. The positive predictive value was 87% and the negative predictive value was 72%. PMID- 15894252 TI - Psychiatry billing for nursing home services: understanding coding options. AB - This article reviews information on coding options, documentation, and payment for both initial and follow-up visits in the nursing home setting. Specific information is provided for both the psychiatry as well as the Evaluation & Management (E&M) code series. Payments are compared for comparable services in order to provide the nursing home psychiatrist with an understanding of the options available. Documentation, background information, and proposed note formatting are also provided. Proper use of the E&M series can provide valuable coding options. PMID- 15894253 TI - Individualizing nutritional care with between-meal snacks for nursing home residents. PMID- 15894254 TI - Information exchange between general practitioner and nursing home physician in The Netherlands. AB - BACKGROUND: In response to the specific characteristics of nursing home residents, the Netherlands has become the only country to develop the specialty of nursing home medicine. The "nursing home physician" has attained independent status. This development has, however, created a division between medical care in the community and medical care in nursing homes, which challenges the quality of the transitional processes taking place when a patient is admitted to or discharged from nursing home care. OBJECTIVES: To give insight into the type of medical information exchanged between general practitioners (GPs) and nursing home physicians (NHPs) at the time of admission, while a patient is under care of the NHP, and at the time of discharge. METHODS: Questionnaires were sent to a sample of 780 GPs, who were selected using a 2-phase sample strategy. Three hypothetical patient vignettes, involving the admission of a patient to a nursing home, to day care, and to an outreaching nursing home care project, were constructed and presented in the questionnaire. GPs were asked to answer questions about the information exchanged during the care of a patient illustrated in each vignette only if they were really familiar with a patient such as presented. The advantage of hypothetical patient vignettes is that each physician reacts to a standardized situation. RESULTS: In the case of admitting a patient to or discharging a patient from the nursing home, results indicate that the continuity of care at those moments will be better ensured if GPs have more frequent personal contacts with NHPs. In the case of day care patients, the study also reveals that GPs who have frequent personal contact with NHPs will share relevant patient information significantly more often at the start of the day care program, both during day care and also when intercurrent medical problems occur. Similar findings can be expected in patients receiving outreaching nursing home care. CONCLUSIONS: The findings indicate the advantages of personal contacts between different medical professionals in exchanging specific patient information. It can be expected that this will lead to more tailor-made medical care for the patient. Adequate exchange of relevant information is an important aspect of mutual collaboration between professionals. Recommendations as to how to achieve more personal contact and a better collaboration among medical professionals are proposed. PMID- 15894256 TI - Is a pressure ulcer a marker for quality of care? PMID- 15894257 TI - The Medicare Modernization Act of 2003, part D. PMID- 15894258 TI - Grass roots efforts: can they make a difference? PMID- 15894259 TI - The role of the immune system in early epithelial carcinogenesis: B-ware the double-edged sword. AB - Cancer is commonly described as a disease of genetic mutations. However, epidemiologic and clinical evidence points to the important but multifaceted role of the host. The immune system has something to say about cancer evolution through promotion of malignancy by inflammatory myeloid cells of the innate immune system. In a report in this issue of Cancer Cell, B cells are implicated as key players in the regulation of chronic inflammation that promotes early events in epithelial carcinogenesis. These are surprising observations, linking antibodies of the adaptive immune system to innate immune responses that drive epithelial carcinogenesis. PMID- 15894260 TI - Chromosomal instability in mouse metastatic pancreatic cancer--it's Kras and Tp53 after all. AB - A human pancreatic cancer progression model from intraepithelial neoplasia to ductal adenocarcinoma has been proposed. This process has been modeled in the mouse by activation of mutant Kras in pancreatic progenitor cells. In this issue of Cancer Cell, present a modification of their initial model by introducing a mutant Tp53. This combination of genetic alterations leads to rapid and increased frequency of neoplasia progression resulting in pancreatic cancers that manifest chromosomal instability in the presence of apparent intact telomeres. These findings introduce Tp53-mediated chromosomal instability as key event for carcinoma development in this mouse model. PMID- 15894261 TI - Considering the critical interface between tumor cells and stromal cells in the search for targets for anticancer therapy. AB - In this issue of Cancer Cell, a paper by Lynch et al. demonstrates how the careful study of changes that occur at the interface between tumor cells and stromal cells led to the discovery of a new function for matrix metalloproteinase 7 (MMP-7) in the formation of osteolytic lesions in prostate cancer. The data suggest that MMP-7 inhibition could be a therapeutic target in prostate cancer. PMID- 15894262 TI - De novo carcinogenesis promoted by chronic inflammation is B lymphocyte dependent. AB - Chronic inflammation predisposes tissue to cancer development; however, regulatory mechanisms underlying recruitment of innate leukocytes toward developing neoplasms are obscure. We report that genetic elimination of mature T and B lymphocytes in a transgenic mouse model of inflammation-associated de novo epithelial carcinogenesis, e.g., K14-HPV16 mice, limits neoplastic progression to development of epithelial hyperplasias that fail to recruit innate immune cells. Adoptive transfer of B lymphocytes or serum from HPV16 mice into T and B cell deficient/HPV16 mice restores innate immune cell infiltration into premalignant tissue and reinstates necessary parameters for full malignancy, e.g., chronic inflammation, angiogenic vasculature, hyperproliferative epidermis. These findings support a model in which B lymphocytes are required for establishing chronic inflammatory states that promote de novo carcinogenesis. PMID- 15894263 TI - Constitutive NF-kappaB activation by the t(11;18)(q21;q21) product in MALT lymphoma is linked to deregulated ubiquitin ligase activity. AB - Mucosa-associated lymphoid tissue (MALT) lymphoma is a common type of lymphoma in extranodal sites. The most frequent chromosome translocation associated with MALT lymphoma is t(11;18)(q21;q21), which generates a chimeric protein of c-IAP2 and MALT1/paracaspase. The c-IAP2/MALT1 fusion protein activates the NF-kappaB pathway, which is considered critical to malignant B cell transformation and lymphoma progression. The mechanism by which this fusion protein activates NF kappaB, however, remains unclear. Here we show that self-oligomerization of the c IAP2/MALT1 protein causes deregulated ubiquitin ligase activity of MALT1/paracaspase. The chimeric protein targets NEMO for polyubiquitination and thereby activates NF-kappaB. Consistent with this finding, NEMO ubiquitination is increased in t(11;18)(q21;q21)-positive MALT lymphoma samples. Thus, t(11;18)(q21;q21) deregulates MALT1/paracaspase ubiquitin ligase activity, causing constitutive NF-kappaB activation and promoting tumorigenesis. PMID- 15894264 TI - Targeting ornithine decarboxylase in Myc-induced lymphomagenesis prevents tumor formation. AB - Checkpoints that control Myc-mediated proliferation and apoptosis are bypassed during tumorigenesis. Genes encoding polyamine biosynthetic enzymes are overexpressed in B cells from E mu-Myc transgenic mice. Here, we report that disabling one of these Myc targets, Ornithine decarboxylase (Odc), abolishes Myc induced suppression of the Cdk inhibitors p21(Cip1) and p27(Kip1), thereby impairing Myc's proliferative, but not apoptotic, response. Moreover, lymphoma development was markedly delayed in E mu-Myc;Odc(+/-) transgenic mice and in E mu Myc mice treated with the Odc inhibitor difluoromethylornithine (DFMO). Strikingly, tumors ultimately arising in E mu-Myc;Odc(+/-) transgenics lacked deletions of Arf, suggesting that targeting Odc forces other routes of transformation. Therefore, Odc is a critical Myc transcription target that regulates checkpoints that guard against tumorigenesis and is an effective target for cancer chemoprevention. PMID- 15894265 TI - Deregulated BCL6 expression recapitulates the pathogenesis of human diffuse large B cell lymphomas in mice. AB - Diffuse large B cell lymphomas (DLBCL) derive from germinal center (GC) B cells and display chromosomal alterations deregulating the expression of BCL6, a transcriptional repressor required for GC formation. To investigate the role of BCL6 in DLBCL pathogenesis, we have engineered mice that express BCL6 constitutively in B cells by mimicking a chromosomal translocation found in human DLBCL. These mice display increased GC formation and perturbed post-GC differentiation characterized by a decreased number of post-isotype switch plasma cells. Subsequently, these mice develop a lymphoproliferative syndrome that culminates with the development of lymphomas displaying features typical of human DLBCL. These results define the oncogenic role of BCL6 in the pathogenesis of DLBCL and provide a faithful mouse model of this common disease. PMID- 15894266 TI - Rational design of shepherdin, a novel anticancer agent. AB - Anticancer agents that selectively kill tumor cells and spare normal tissues are urgently needed. Here, we engineered a cell-permeable peptidomimetic, shepherdin, modeled on the binding interface between the molecular chaperone Hsp90 and the antiapoptotic and mitotic regulator, survivin. Shepherdin makes extensive contacts with the ATP pocket of Hsp90, destabilizes its client proteins, and induces massive death of tumor cells by apoptotic and nonapoptotic mechanisms. Conversely, shepherdin does not reduce the viability of normal cells, and does not affect colony formation of purified hematopoietic progenitors. Systemic administration of shepherdin in vivo is well tolerated, and inhibits human tumor growth in mice without toxicity. Shepherdin could provide a potent and selective anticancer agent in humans. PMID- 15894267 TI - Trp53R172H and KrasG12D cooperate to promote chromosomal instability and widely metastatic pancreatic ductal adenocarcinoma in mice. AB - To define the genetic requirements for pancreatic ductal adenocarcinoma (PDA), we have targeted concomitant endogenous expression of Trp53(R172H) and Kras(G12D) to the mouse pancreas, revealing the cooperative development of invasive and widely metastatic carcinoma that recapitulates the human disease. The primary carcinomas and metastases demonstrate a high degree of genomic instability manifested by nonreciprocal translocations without obvious telomere erosion-hallmarks of human carcinomas not typically observed in mice. No mutations were discovered in other cardinal tumor suppressor gene pathways, which, together with previous results, suggests that there are distinct genetic pathways to PDA with different biological behaviors. These findings have clear implications for understanding mechanisms of disease pathogenesis, and for the development of detection and targeted treatment strategies. PMID- 15894268 TI - MMP-7 promotes prostate cancer-induced osteolysis via the solubilization of RANKL. AB - We developed a rodent model that mimics the osteoblastic and osteolytic changes associated with human metastatic prostate cancer. Microarray analysis identified MMP-7, cathepsin-K, and apolipoprotein D as being upregulated at the tumor-bone interface. MMP-7, which was produced by osteoclasts at the tumor-bone interface, was capable of processing RANKL to a soluble form that promoted osteoclast activation. MMP-7-deficient mice demonstrated reduced prostate tumor-induced osteolysis and RANKL processing. This study suggests that inhibition of MMP-7 will have therapeutic benefit in the treatment of prostate cancer-induced osteolysis. PMID- 15894269 TI - The alphabeta versus gammadelta T cell fate decision: when less is more. PMID- 15894270 TI - T cell expansion: the survivin interface between cell proliferation and cell death. PMID- 15894271 TI - Dipping into the cytosol to broaden the MHC class II peptide repertoire. PMID- 15894272 TI - Phagocytosis: elegant complexity. AB - Phagocytosis requires receptor-mediated recognition of particles, usually in the guise of infectious agents and apoptotic cells. Phagosomes fuse with lysosomes to generate phagolysosomes, which play a key role in enzymatic digestion of the internalized contents into component parts. Recent findings indicate that a simple paradigm of a single cognate receptor interaction that guides the phagosome to phagolysosome formation belies the complexity of combinatorial receptor recognition and diversity of phagosome function. In fact, phagosomes are comprised of hundreds of proteins that play a key role in deciphering the contents of the phagosome and in defining host response. In this review we discuss how the challenge of recognizing diverse molecular patterns is met by combinatorial interactions between phagocytic receptors. Furthermore, these combinations are dynamic and both sculpt the balance between a proinflammatory or anti-inflammatory response and direct phagosome diversity. We also indicate an important role for genetically tractable model organisms in defining key components of this evolutionarily conserved process. PMID- 15894273 TI - Complexity and complementarity of outer membrane protein A recognition by cellular and humoral innate immunity receptors. AB - Outer membrane protein A (OmpA) is a conserved major component of the outer membrane of Enterobacteriaceae. Here, we report that OmpA from Klebsiella pneumoniae (KpOmpA) activates macrophages and dendritic cells (DCs) in a TLR2 dependent way. However, TLR2 does not account for binding of KpOmpA to innate immune cells. KpOmpA binds the scavenger receptors (SRs) LOX-1 and SREC-I, but not other members of the same family. LOX-1 colocalizes and cooperates with TLR2 in triggering cellular responses. The TLR2-activated functional program includes production of the long pentraxin PTX3, a soluble pattern recognition receptor involved in resistance against diverse pathogens. PTX3, in turn, binds KpOmpA but does not affect recognition of this microbial moiety by cellular receptors. KpOmpA-elicited in vivo inflammation is abrogated in TLR2(-/-) mice and significantly reduced in PTX3(-/-) mice. Thus, SR-mediated KpOmpA recognition and TLR2-dependent cellular activation set in motion a nonredundant PTX3-mediated humoral amplification loop of innate immunity. PMID- 15894274 TI - Dendritic cells maximize the memory CD8 T cell response to infection. AB - Costimulatory signals from dendritic cells (DCs) are required for naive T cells to respond to antigenic stimulation. To what extent DCs reactivate memory T cells during recall responses is not known. Here, an in vivo depletion system has been used to analyze the role of DCs in reactivating CD8 memory T cells during recall responses to three different microbial infections. We show a profound decrease in the numbers of responding memory CD8 T cells in both lymphoid and nonlymphoid tissues during the recall responses to infection with vesicular stomatitis virus, Listeria monocytogenes (Lm), or influenza virus. These data show that interaction with DCs is a major mechanism driving T cell reactivation in vivo, even during a tissue-specific infection of the respiratory tract. PMID- 15894275 TI - Lamp-2a facilitates MHC class II presentation of cytoplasmic antigens. AB - Extracellular antigens are internalized and processed before binding MHC class II molecules within endosomal and lysosomal compartments of professional antigen presenting cells (APC) for subsequent presentation to T cells. Yet select cytoplasmic peptides derived from autoantigens also intersect and bind class II molecules via an unknown mechanism. In human B lymphoblasts, inhibition of the peptide transporter associated with antigen processing (TAP) failed to alter class II-restricted cytoplasmic epitope presentation. By contrast, decreased display of cytoplasmic epitopes via class II molecules was observed in cells with diminished expression of the lysosome-associated membrane protein-2 (Lamp-2). Overexpression of Lamp-2 isoform A (Lamp-2a), an established component of chaperone-mediated autophagy, enhanced cytoplasmic autoantigen presentation. Manipulating APC expression of heat shock cognate protein 70 (hsc70), a cofactor for Lamp-2a, also altered cytoplasmic class II peptide presentation. These results demonstrate a novel role for the lysosomal Lamp-2a-hsc70 complex in promoting immunological recognition and antigen presentation. PMID- 15894276 TI - TCR signal strength influences alphabeta/gammadelta lineage fate. AB - Signals transduced by T cell antigen receptors (TCRs) have been shown to be critical for alphabeta and gammadelta T cell development, but their role in lineage determination remains poorly defined. Two models have been forwarded for alphabeta/gammadelta lineage choice: the instructive model and the stochastic model. Recent data, however, are inconsistent with either model. In this study, we devised an experimental system in which lineage fate was controlled exclusively by the gammadeltaTCR. We then analyzed the impact of TCR signal strength on alphabeta/gammadelta lineage development by altering the surface expression or signaling potential of the gammadeltaTCR complex. We found that increasing the gammadeltaTCR signal strength favored gammadelta lineage development, whereas weakening the gammadeltaTCR signal favored alphabeta lineage development. These results support a model in which the strength of the TCR signal is a critical determinant in the lineage fate decision. PMID- 15894277 TI - Attenuation of gammadeltaTCR signaling efficiently diverts thymocytes to the alphabeta lineage. AB - The role of the T cell antigen receptor complex (TCR) in alphabeta/gammadelta lineage commitment remains controversial, in particular whether different TCR isoforms intrinsically favor adoption of a certain lineage. Here, we demonstrate that impairing the signaling capacity of a gammadeltaTCR complex enables it to efficiently direct thymocytes to the alphabeta lineage. In the presence of a ligand, a transgenic gammadeltaTCR mediates almost exclusive adoption of the gammadelta lineage, while in the absence of ligand, the same gammadeltaTCR promotes alphabeta lineage development with efficiency comparable to the pre-TCR. Importantly, attenuating gammadeltaTCR signaling through Lck deficiency causes reduced ERK1/2 activation and Egr expression and diverts thymocytes to the alphabeta lineage even in the presence of ligand. Conversely, ectopic Egr overexpression favors gammadelta T cell development. Our data support a model whereby gammadelta versus alphabeta lineage commitment is controlled by TCR signal strength, which depends critically on the ERK MAPK-Egr pathway. PMID- 15894278 TI - Evidence that HIV-1 encodes an siRNA and a suppressor of RNA silencing. AB - In plants and invertebrate animals, RNA silencing is a form of nucleic acid-based adaptive immunity. By contrast, jawed vertebrates have evolved complex protein based adaptive immunity. Although short interfering RNAs (siRNAs) have been used as artificial tools to silence viral infection in human cells, it remains unknown whether mammalian viruses naturally elicit such immunity in vertebral cells. Here, we report the evidence that HIV-1 encodes viral siRNA precursors in its genome and that natural HIV-1 infection provokes nucleic acid-based immunity in human cells. To combat this cellular defense, HIV-1 has evolved in its Tat protein a suppressor of RNA silencing (SRS) function. Tat abrogates the cell's RNA-silencing defense by subverting the ability of Dicer to process precursor double-stranded RNAs into siRNAs. PMID- 15894279 TI - Sustained survivin expression from OX40 costimulatory signals drives T cell clonal expansion. AB - Sustained signaling from the T cell receptor (TCR) and costimulatory molecules is thought necessary for generating high numbers of effector T cells. Here, we show that Survivin is controlled in peripheral T cells by OX40 cosignaling via sustained PI3k and PKB activation. Survivin is induced by OX40 independent of mitotic progression in late G1, and blocking Survivin suppresses S-phase transition and division of T cells and leads to apoptosis. Moreover, Survivin expression alone is sufficient to restore proliferation and to antagonize apoptosis in costimulation-deficient T cells and can rescue T cell expansion in vivo. Survivin allows effector T cells to accumulate in large numbers, but Bcl-2 family proteins are required for T cell survival after the phase of active division. Thus, sustained Survivin expression from costimulatory signaling maintains T cell division over time and regulates the extent of clonal expansion. PMID- 15894280 TI - GCN2 kinase in T cells mediates proliferative arrest and anergy induction in response to indoleamine 2,3-dioxygenase. AB - Indoleamine 2,3 dioxygenase (IDO) catabolizes the amino acid tryptophan. IDO expressing immunoregulatory dendritic cells (DCs) have been implicated in settings including tumors, autoimmunity, and transplant tolerance. However, the downstream molecular mechanisms by which IDO functions to regulate T cell responses remain unknown. We now show that IDO-expressing plasmacytoid DCs activate the GCN2 kinase pathway in responding T cells. GCN2 is a stress-response kinase that is activated by elevations in uncharged tRNA. T cells with a targeted disruption of GCN2 were not susceptible to IDO-mediated suppression of proliferation in vitro. In vivo, proliferation of GCN2-knockout T cells was not inhibited by IDO-expressing DCs from tumor-draining lymph nodes. IDO induced profound anergy in responding wild-type T cells, but GCN2-knockout cells were refractory to IDO-induced anergy. We hypothesize that GCN2 acts as a molecular sensor in T cells, allowing them to detect and respond to conditions created by IDO. PMID- 15894281 TI - Dynamics and function of Langerhans cells in vivo: dermal dendritic cells colonize lymph node areas distinct from slower migrating Langerhans cells. AB - Langerhans cells (LCs) are prominent dendritic cells (DCs) in epithelia, but their role in immunity is poorly defined. To track and discriminate LCs from dermal DCs in vivo, we developed knockin mice expressing enhanced green fluorescent protein (EGFP) under the control of the langerin (CD207) gene. By using vital imaging, we showed that most EGFP(+) LCs were sessile under steady state conditions, whereas skin inflammation induced LC motility and emigration to lymph nodes (LNs). After skin immunization, dermal DCs arrived in LNs first and colonized areas distinct from slower migrating LCs. LCs reaching LNs under steady state or inflammatory conditions expressed similar levels of costimulatory molecules. Langerin and EGFP were also expressed on thymic DCs and on blood derived, CD8alpha(+) DCs from all secondary lymphoid organs. By using a similar knockin strategy involving a diphtheria toxin receptor (DTR) fused to EGFP, we demonstrated that LCs were dispensable for triggering hapten-specific T cell effectors through skin immunization. PMID- 15894282 TI - Heterocoagulation of chalcopyrite and pyrite minerals in flotation separation. AB - Heterocoagulation between various fine mineral particles contained within a mineral suspension with different structural and surface chemistry can interfere with the ability of the flotation processes to selectively separate the minerals involved. This paper examines the interactions between chalcopyrite (a copper mineral) and pyrite (an iron mineral often bearing gold) as they approach each other in suspensions with added chemicals, and relates the results to the experimental data for the flotation recovery and selectivity. The heterocoagulation was experimentally studied using the electrophoretic light scattering (ELS) technique and was modelled by incorporating colloidal forces, including the van der Waals, electrostatic double layer and hydrophobic forces. The ELS results indicated that pyrite has a positive zeta potential (zeta) up to its isoelectric point (IEP) at approximately pH 2.2, while chalcopyrite has a positive zeta up to its IEP at approximately pH 5.5. This produces heterocoagulation of chalcopyrite with pyrite between pH 2.2 and pH 5.5. The heterocoagulation was confirmed by the ELS spectra measured with a ZetaPlus instrument from Brookhaven and by small-scale flotation experiments. PMID- 15894283 TI - Interfacial rheology of adsorbed layers with surface reaction: on the origin of the dilatational surface viscosity. AB - A theoretical study of the phenomena, occurring in an adsorbed layer, subject to small dilatational perturbations was carried out. Two main processes, provoked by the perturbations (surface reaction and surfactant transport onto the surface) were considered. The reaction was described by means of the reaction coordinate. The derived general rheological equation for insoluble surfactants, gave as limiting cases Voight and Maxwell type equations for fast and slow reactions, respectively. Expressions for all characteristics of the process (surface elasticity, reaction elasticity, reaction relaxation time and dilatational surface viscosity) were obtained. The obtained generalized rheological equation for reactions involving soluble surfactants is a dynamic analog of Gibbs adsorption isotherms for a multi-component system with surface reaction, since similarly to Gibbs equation it relates the surface stress only to surface variables. It gives as limiting cases generalized forms for soluble surfactants of Voight and Maxwell equations. All new rheological equations were analyzed for deformations with constant rate and periodic oscillations and they were applied to three simple surface reactions (monomolecular with one product, dimerization and association). The mass transfer was analyzed initially in the absence of surface reaction. In this system the surface stress is purely elastic, but it was shown that if the adsorption perturbation is small, regardless of the type of surface perturbation and the mechanism of adsorption, the process of mass transport always obeys a Maxwell type rheological equation. For all considered processes surface viscosities were defined, but they were called "apparent", because they stem from diffusion, rather than from interaction between the surfactant molecules and they depend not only on surface parameters, but also on the geometry of the system. The often used in the literature correlations between the lifetime of emulsions and foams and the imaginary elasticity were analyzed. It was shown that this approach lacks serious scientific foundations and could lead to erroneous conclusions. Finally, the problem for the coupling of the surfactant diffusion with the chemical reaction was analyzed and it was demonstrated on a simple example how it could be tackled. PMID- 15894284 TI - Impact of railroad ballast type on frontal plane ankle kinematics during walking. AB - Five healthy male subjects walked on a control surface (level concrete), and two sloped rock surfaces (walking ballast-rock about 1.9 cm across; main line ballast rock about 3.8 cm across) while their rearfoot motion (defined throughout as ankle inversion/eversion as seen from the frontal plane) was measured to determine if the different walking surfaces caused different ankle kinematics. The ballast was placed in 5m long trays that were tilted 7 degrees in the transverse plane. Rearfoot motion was measured while the subjects walked the length of the respective surfaces wearing work boots. A repeated measures ANOVA and a subsequent multiple comparison test revealed that the rearfoot range of motion was significantly greater walking on the main line ballast than walking on either the walking ballast or the level concrete. Meanwhile, the mean range of rearfoot motion for walking ballast was not significantly different from that resulting from walking on concrete. Variability was more than twice as great walking on main line ballast than walking on level concrete. Rearfoot angular velocities walking on level concrete and walking ballast were not significantly different, but both were significantly less than walking on main line ballast. Results suggested that rearfoot motion could be reduced if railroads placed walking ballast where trainmen have to walk as part of their jobs. PMID- 15894285 TI - Parallel development of cardiomyocytes and neurons in embryonic stem cell culture. AB - Recent studies suggest that there are strong parallels between development and patterning of the vertebrate vascular system and the nervous system. While previous observations reported generation of vascular and neuronal progenitors from embryonic stem (ES) cells, the question of parallel development of vascular and neuronal cells in the same culture has not yet been investigated. Mouse D3 ES cells were cultured for 4 days in differentiation medium IMDM with 15% FBS in 100 mm non-adhesive Petri dishes to allow cells to aggregate and form embryoid bodies. At day 5, fibronectin or all-trans retinoic acid with fibronectin was added to the culture. On day 9, the embryoid bodies were seeded on poly-L ornithine/fibronectin-coated plates. After plating, half of the plates were treated with laminin for 3 days and maintained for 1 week in Neurobasal media with B27. Here we show that ES cells differentiate into interconnected rhythmically contracting aggregates of functional cardiomyocytes and neurons. Double immunofluorescence with anti-phospholamban, anti-SERCA2 antibodies to detect cardiomyocytes and with anti-MAP2 antibodies to detect neurons revealed the cell aggregates consisting entirely of cardiomyocytes with neuronal cells located on the periphery or covering the aggregate's surface. The observed concurrent development of cardiomyocytes and neurons suggests bidirectional communication between both cell types. We propose that crosstalk between cardiovascular and neuronal progenitors is an important mechanism for the development of both systems. PMID- 15894286 TI - Maturation efficiency, trypsin sensitivity, and optical properties of Arg96, Glu222, and Gly67 variants of green fluorescent protein. AB - Spontaneous chromophore biosynthesis in green fluorescent protein (GFP) is initiated by a main-chain cyclization reaction catalyzed by the protein fold. To investigate the structural prerequisites for chromophore formation, we have substituted the conserved residues Arg96, Glu222, and Gly67. Upon purification, the variants can be ordered based on their decreasing extent of chromophore maturation according to the series EGFP, E222Q, R96K, G67A, and R96M. Arg96 and Glu222 appear to play catalytic roles, whereas Gly67 is likely important in interior packing to enforce correct hydrogen bonding to Arg96. The effect of Arg96 can be partially compensated for by a lysine, but not by a methionine residue, confirming its electrophilic role. Limited trypsinolysis data suggest that protein stability is largely unaffected by the presence of the chromophore, inconsistent with the mechanical compression hypothesis. Trends in optical properties may be related to the degree of chromophore charge delocalization, which is modulated by residue 96. PMID- 15894287 TI - Magic roundabout, a tumor endothelial marker: expression and signaling. AB - Molecular signals that guide blood vessels to specific paths are not fully deciphered, but are thought to be similar to signals that mediate neuronal guidance. These cues are not only critical for normal blood vessel development, but may also play a major role in tumor angiogenesis. In this study, we have demonstrated the tumor endothelial specific expression of a Robo family member, magic roundabout (MRB), functionally characterized its role in endothelial cell migration and defined a signaling pathway that might mediate this function. We show that MRB is differentially over-expressed in tumor endothelial cells versus normal adult endothelial cells in numerous solid tumors. Moreover, over expression of MRB in endothelial cells activates MRB in a ligand-independent fashion, and activation of MRB via Slit2, a putative ligand, results in inhibition of VEGF and FGF induced migration. We also demonstrate that MRB induced inhibition of endothelial migration is partially mediated by the Ras-Raf Mek-Erk signaling pathway. We therefore hypothesize that expression of MRB is involved in regulating the migration of endothelial cells during tumor angiogenesis. PMID- 15894288 TI - Plant K(in) and K(out) channels: approaching the trait of opposite rectification by analyzing more than 250 KAT1-SKOR chimeras. AB - Members of the Shaker-like plant K(+) channel family share a common structure, but are highly diverse in their function: they behave as either hyperpolarization activated inward-rectifying (K(in)) channels, or leak-like (K(weak)) channels, or depolarization-activated outward-rectifying (K(out)) channels. Here we created 256 chimeras between the K(in) channel KAT1 and the K(out) channel SKOR. The chimeras were screened in a potassium-uptake deficient yeast strain to identify those, which mediate potassium inward currents, i.e., which are functionally equivalent to KAT1. This strategy allowed us to identify three chimeras which differ from KAT1 in three parts of the polypeptide: the cytosolic N-terminus, the cytosolic C-terminus, and the putative voltage-sensor S4. Additionally, mutations in the K(out) channel SKOR were generated in order to localize molecular entities underlying its depolarization activation. The triple mutant SKOR-D312N-M313L I314G, carrying amino-acid changes in the S6 segment, was identified as a channel which did not display any rectification in the tested voltage-range. PMID- 15894289 TI - Enhanced expression from the human cytomegalovirus immediate-early promoter in a non-replicating adenovirus encoded reporter gene following cellular exposure to chemical DNA damaging agents. AB - We have examined expression from the human cytomegalovirus (CMV) promoter of a reporter gene encoded in a replication-deficient adenovirus following cellular exposure to heat shock and chemical DNA damaging agents. Expression of the reporter gene was enhanced following prior treatment of cells with cisplatin and N-acetoxy-acetylaminofluorine, but not heat shock. This enhancement was more pronounced and induced by lower chemical concentrations in xeroderma pigmentosum (XP) and Cockayne syndrome fibroblasts that are deficient in the transcription coupled repair (TCR) pathway of nucleotide excision repair (NER) compared to that in TCR-proficient XP-C and normal strains. This is consistent with an induction of expression from the CMV promoter mediated by persistent (unrepaired) DNA damage in active genes. We show also that expression of the CMV-driven reporter is enhanced following treatment of several human tumour cell lines. This later finding has implications for combined chemotherapy and gene therapy using CMV driven expression vectors. PMID- 15894290 TI - A role for manganese superoxide dismutase in apoptosis after photosensitization. AB - The oxidative stress triggered by photodynamic therapy (PDT) involves generation of cytotoxic reactive oxygen species, including superoxide radical, accumulation of de novo-generated ceramide, and induction of apoptosis. Since PDT with the photosensitizer phthalocyanine Pc 4 induces mitochondrial damage and the superoxide scavenger manganese superoxide dismutase (MnSOD) is localized to mitochondria, here we tested genetically the role of MnSOD in apoptosis and ceramide accumulation after photosensitization with Pc 4. Jurkat cells overexpressing wild-type MnSOD were protected from Pc 4-PDT-initiated apoptosis, but not from increased ceramide response to Pc 4-PDT. In Jurkat cells overexpressing mutant MnSOD, however, DEVDase activation and ceramide formation were promoted post-Pc 4-PDT. Similarly, in MnSOD-null cells, Pc 4-PDT-induced apoptosis, as well as ceramide accumulation, were enhanced compared to their normal counterparts. The data show that MnSOD affects sensitivity of cells to Pc 4-PDT-initiated apoptosis, and partly ceramide accumulation, suggesting that the processes are superoxide-mediated. PMID- 15894291 TI - Licofelone, a novel 5-LOX/COX-inhibitor, attenuates leukocyte rolling and adhesion on endothelium under flow. AB - The main mechanism of action of non-steroidal anti-inflammatory drugs (NSAIDs) is the inhibition of cycloxygenases COX-1 and COX-2. During recent years, combined 5 LOX/COX-inhibition, interfering with the biosynthesis of both prostaglandins and leukotrienes (LTs), has emerged as a possibility to avoid side effects related to COX-inhibition. The aim of the present study was to investigate if there is a contribution of mechanisms other than the reduction of inflammatory prostaglandins and leukotrienes to the anti-inflammatory effect of the LOX/COX inhibitor licofelone. In a flow chamber assay, licofelone (10-30 microM) dose dependently decreased both the rolling and adhesion of leukocytes on endothelial cells (EC). In contrast, no effects were found after treatment of EC with the unselective COX-1/COX-2 inhibitor indomethacin (30 microM), the potent and selective 5-LOX inhibitor, ZD-2138 (30 microM), the mainly COX-2 inhibitor aceclofenac (30 microM), the selective COX-2 inhibitor celecoxib (30 microM) and the combination of ZD-2138 with the selective COX-2 inhibitor celecoxib (30 microM). In the presence of licofelone (30 microM) the expression of E-selectin mRNA in cytokine-stimulated EC was attenuated, whereas no NSAID (30 microM) tested showed any effect on E-selectin expression. Moreover, licofelone treatment (30 microM) attenuated expression of VCAM-1 and ICAM-1 on inflammatory EC. The effect of licofelone on leukocyte recruitment was also evaluated in vivo. Using a mouse peritonitis model it was found that leukocyte accumulation was markedly reduced in licofelone treated animals (100mg/kg) compared to untreated mice. Thus, the novel 5-LOX/COX inhibitor licofelone possesses anti-inflammatory activity that, in addition to COX/LOX inhibition, involves effects on leukocyte endothelial interactions. PMID- 15894292 TI - Reactive oxygen species generation and its role in the differential cytotoxicity of the arylhydroxylamine metabolites of sulfamethoxazole and dapsone in normal human epidermal keratinocytes. AB - Cutaneous drug reactions (CDR) are responsible for numerous minor to life threatening complications. Though the exact mechanism for CDR is not completely understood, evidence suggests that bioactivation of drugs to reactive oxygen or nitrogen species is an important factor in the initiation of these reactions. Several CDR-inducing drugs having an arylamine functional group, such as sulfamethoxazole (SMX) and dapsone (DDS), undergo bioactivation to reactive arylhydroxylamine metabolites. These metabolites can generate cellular oxidative stress by forming reactive oxygen species (ROS). Several studies have demonstrated a higher cytotoxicity with DDS hydroxylamine (DDS-NOH) compared to SMX hydroxylamine (SMX-NOH). To investigate the role of differential ROS generation in the higher cytotoxicity of DDS-NOH, hydroxylamine metabolites of SMX and DDS were synthesized and ROS formation by these metabolites determined. DDS-NOH and its analogues/metabolites consistently resulted in higher ROS formation as compared to SMX-NOH. However, comparison of the ROS generation and cytotoxicity of a series of arylhydroxylamine analogues of DDS did not support a simple correlation between ROS generation and cell death. Numerous ROS scavengers were found to reduce metabolite-induced ROS formation, with differences in the potency between the agents. The decrease in DDS-NOH-induced ROS generation in NHEK with ascorbic acid, N-acetylcysteine, Trolox, and melatonin was 87, 86, 44, and 16%, respectively. Similarly, the cytotoxicity and adduct formation of DDS NOH in NHEK was reduced in the presence of ascorbic acid. In summary, these studies show that arylhydroxylamine metabolites of SMX/DDS induce ROS generation in NHEK, though such generation is not directly related to cytotoxicity. PMID- 15894293 TI - Inhibition of the mitochondrial ATP synthesis by polygodial, a naturally occurring dialdehyde unsaturated sesquiterpene. AB - Polygodial is a naturally occurring sesquiterpene dialdehyde that exhibits several pharmacologically interesting activities. Among them, its antifungal properties have been more thoroughly studied. The mitochondrial ATPase has been suggested as one of the possible targets for polygodial action. However, its mechanism of action is not well defined yet. The effect of polygodial on the mitochondrial energy metabolism is described in this paper. Polygodial inhibited ATP synthesis coupled to succinate oxidation in beef-heart submitochondrial particles at concentrations (IC(50)=2.4+/-0.1 microM) which marginally affected electron transport and ATPase activity (IC(50)=97+/-4 microM). A transitory stimulation of the electron transport in intact rat liver mitochondria in state 4 was also obtained at low polygodial concentrations (EC(50)=20+/-4 microM). These results suggest that polygodial uncouples ATP synthesis from electron transport at low concentrations. Similar concentrations of polygodial partially abolished the ANS fluorescence enhancement (IC(50)=2.2+/-0.4 microM) induced by succinate oxidation in submitochondrial particles but did not collapse the DeltapH. We postulate that polygodial uncouples mitochondrial ATP synthesis by affecting the electrical properties of the membrane surface and consequently collapsing the membrane potential (Deltapsi) and/or the localized transmembrane pH difference (DeltapH(S)) without affecting the DeltapH between the two bulk aqueous phases (DeltapH(B)). The relevance of these findings for the understanding of the biochemical basis of the antifungal activity of polygodial and the evaluation of its potentiality as a therapeutic agent are discussed. PMID- 15894294 TI - Site specific alterations of adipose tissue mitochondria in 3'-azido-3' deoxythymidine (AZT)-treated rats: an early stage in lipodystrophy? AB - Although it is well accepted that treatment with nucleoside reverse transcriptase inhibitors (NRTIs) modifies fat metabolism and fat distribution in humans, the mechanisms underlying these modifications are not yet known. The present investigation examines the effects of chronic oral administration of 3'-azido-3' deoxythymidine (AZT) on the mitochondrial metabolism and the redox status management of rat white adipose tissues originating from two anatomical sites, as well as of the rat liver. Results showed that AZT treatment induced differential effects on the mitochondrial functions depending on the anatomical localisation. Indeed, in inguinal adipose tissue, a significant decrease in the cytochrome c oxidase activity and in the mitochondrial DNA (mtDNA) content was observed, whereas the activity of citrate synthase, a mitochondrial protein exclusively encoded by the nucleus, was not affected. In contrast, no significant change in these parameters could be detected for epididymal tissue and for liver. In parallel, no oxidative stress could be detected after treatment, for both white adipose tissues and for liver, even though treated liver exhibited several modifications in redox management. Taken together, these data demonstrate differential mitochondrial effects of AZT on subcutaneous versus visceral white adipose tissue. Moreover, the decrease in mitochondrial oxidative capacity of inguinal adipocyte consecutive to AZT treatment is not primarily due to an oxidative stress per se, but rather to a depletion of the mtDNA content per cell. PMID- 15894295 TI - Cell apoptosis induced by a synthetic carbazole compound LCY-2-CHO is mediated through activation of caspase and mitochondrial pathways. AB - The mechanisms involved in the apoptotic effect of LCY-2-CHO [9-(2-chlorobenzyl) 9H-carbazole-3-carbaldehyde], a synthetic carbazole derivative identified as an anti-inflammatory compound, were studied. Cell cycle analysis by propidium iodide staining in human THP-1 monocytic leukemia cells showed the ability of LCY-2-CHO to increase cell population in sub-G1 stage with time- and concentration dependent manners. LCY-2-CHO-mediated cell death was also demonstrated by DNA laddering and was not related to the release of lactate dehydrogenase. Apoptosis in THP-1 cells induced by LCY-2-CHO was accompanied by the Bid cleavage, collapse of mitochondrial transmembrane potential, the release of cytochrome c and the activation of caspase-3. The apoptotic effect of LCY-2-CHO was diminished by the presence of zVEID-fmk (caspase-6 inhibitor), zIETD-fmk (caspase-8 inhibitor), and zVAD-fmk (non-selective caspase inhibitor), but was not altered by several antioxidants, and cathepsin inhibitor. The Bid cleavage and loss of mitochondrial transmembrane potential, but not the cytochrome c release, were reversed by zIETD fmk. Comparing the cell selectivity of LCY-2-CHO, we found T-cell acute lymphoblastic CEM leukemia cells were sensitive to 1 microM LCY-2-CHO, acute myeloid leukemia HL-60 cells underwent apoptosis at 10 microM, while adherent cancer cells, such as PC3, HT29 and MCF-7, were resistant to 30 microM LCY-2-CHO within 24-h incubation. Taken together in the present study, we demonstrated LCY 2-CHO might be apoptotic for malignant hematopoietic cells but not anchorage dependent cells. This action is mediated by an intrinsic caspase-dependent apoptotic event involving mitochondria. PMID- 15894296 TI - Functional expression of particular isoforms of excitatory amino acid transporters by rodent cartilage. AB - In the present study, we have attempted to demonstrate functional expression by the rodent cartilage of particular isoforms of excitatory amino acid transporters (EAATs) essentially required for central glutamatergic signal termination. Constitutive expression of mRNA was shown for the first time with the neuronal EAAT subtype excitatory amino acid carrier-1 (EAAC1), in addition to glial subtypes such as glutamate aspartate transporter (GLAST) and glutamate transporter-1 (GLT-1), in rat costal chondrocytes cultured for 7-21 days on reverse transcription polymerase chain reaction (RT-PCR). Western blotting analysis confirmed the expression of corresponding proteins for both GLAST and GLT-1 in cultured chondrocytes. The accumulation of [(3)H]glutamate (Glu) occurred in a temperature- and sodium-dependent manner with biochemical and pharmacological profiles similar to those seen for brain EAATs in chondrocytes cultured for 7 days, while [(3)H]Glu accumulation consisted of a single component with a K(m) of 39.1+/-2.3 microM and a V(max) of 1320+/-120pmol/mg protein/min, respectively. In organotypic cultured metatarsals isolated before vascularization from embryonic mice, where cells underwent maturational development from resting to proliferating, prehypertrophic, hypertrophic and calcified chondrocytes in a progressive order of cellular differentiation, moreover, mRNA expression was seen for GLAST, GLT-1 and EAAT4 but not for EAAC1 subtypes. Immunohistochemical analysis revealed distribution profiles different from each other with GLAST, GLT 1 and EAAT4 isoforms in sections of cultured metatarsals and isolated tibiae. These results suggest that extracellular Glu could be cleared up into intracellular locations through particular glial and/or neuronal EAAT isoforms functionally expressed by the rodent cartilage. PMID- 15894297 TI - Serotonergic hyperinnervation into the dopaminergic denervated striatum compensates for dopamine conversion from exogenously administered l-DOPA. AB - The aim of this study was to determine whether raphe-striatal serotonergic neurons of adult rats with extensive nigro-striatal dopaminergic denervation are induced by injection of exogenous L-DOPA to contain dopamine. Double-labeling immunofluorescence study was conducted. In the lesioned striatum of rats that received L-DOPA, serotonergic hyperinnervation was observed, and dopamine was detected in serotonergic varicose fibers. These findings suggest that striatal serotonergic hyperinnervation can compensate for the lost function of dopaminergic neurons. PMID- 15894298 TI - Mycobacterium bovis BCG induces CXC chemokine ligand 8 secretion via the MEK dependent signal pathway in human epithelial cells. AB - Current knowledge of the cellular signaling by Mycobacterium bovis bacillus Calmette-Guerin (BCG) in epithelial cells is still limited. In this study, we provide evidence that the signaling events induced by M. bovis BCG in these cells included phosphorylation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK). Our data also demonstrate that M. bovis BCG-induced CXC chemokine ligand (CXCL)8 release in epithelial cells was reduced by a mitogen-activated protein/ERK kinase (MEK) inhibitor (PD98059), but not by a p38 MAPK (SB203580) inhibitor. In addition, we found that a second and more potent MEK inhibitor (U0126) significantly blocked CXCL8 release in epithelial cells by M. bovis BCG. Evaluation of CXCL8 RNA messages by reverse transcription polymerase chain reaction (RT-PCR) revealed that the inhibitory effect of PD98059 and U0126 was associated with a reduction in this parameter. Moreover, the induction of CXCL8 secretion in epithelial cells by M. bovis BCG occurs at the transcription level. Collectively, the findings reported in the present study suggest that MEK signaling is essential for the induction of CXCL8 in epithelial cells in response to M. bovis BCG. PMID- 15894299 TI - Fatty acid methyl esters are detectable in the plasma and their presence correlates with liver dysfunction. AB - BACKGROUND: Methanol is a component of certain alcoholic beverages and is also an endogenously formed product. On this basis, we have proposed that methanol may promote synthesis of fatty acid methyl esters (FAMEs) in the same way that ethanol promotes fatty acid ethyl ester (FAEE) synthesis. We tested the hypothesis that FAMEs appear in the blood after ethanol intake. METHODS: Patient plasma samples obtained from our laboratory (n=78) were grouped according to blood ethanol concentrations (intoxicated, blood ethanol >800 mg/l) and non intoxicated. These samples were further subdivided into groups based on whether the patient had normal or abnormal liver function tests (abnormal, defined as > or =1 abnormality of plasma alanine and aspartate aminotransferase, albumin, total bilirubin, and alkaline phosphatase). A separate set of plasma samples were also divided into normal and abnormal groups based on pancreatic function tests (amylase and lipase). There were no patients with detectable ethanol in this group. Patients with abnormalities in pancreatic function tests were included upon recognition of endogenously produced FAMEs by patients with liver function test abnormalities. FAMEs were extracted from plasma and individual species of FAMEs quantified by gas chromatography-mass spectrometry (GC/MS). RESULTS: Increased concentrations of FAME were found in patient samples with evidence of liver dysfunction, regardless of whether or not they were intoxicated (n=21, p=0.01). No significant differences in plasma FAME concentrations were found between patients with normal (n=15) versus abnormal pancreatic function tests (n=22, p=0.72). CONCLUSIONS: The presence of FAMEs in human plasma may be related to the existence of liver disease, and not to blood ethanol concentrations or pancreatic dysfunction. The metabolic pathways associated with FAME production in patients with impaired liver function remain to be identified. PMID- 15894300 TI - A Rwandan altitude blood gas, acid-base and hemoglobin study. PMID- 15894301 TI - Laboratory screening of connective tissue diseases by a new automated ENA screening assay (EliA Symphony) in clinically defined patients. AB - BACKGROUND: The measurement of antinuclear antibodies (ANA) is used in the autoimmune laboratory for the screening of connective tissue diseases (CTD). ANA measurements are mainly performed by indirect immunofluorescence (IIF) on HEp-2 cells or by enzyme immunoassay (EIA). The objective of this study was to clinically evaluate an automated EIA for extractable nuclear antigens (ENA) which lacks anti-dsDNA for the screening of CTD. METHODS: The study involved a total of 170 serum samples, 54 from patients with CTD, 26 from patients with other autoimmune diseases, and 90 from patients with non-autoimmune diseases. For all sera, ANA detection was performed by IIF and by EliA Symphony (Pharmacia Diagnostics, Freiburg, Germany), an ENA screening which detects the following autoantibodies: SSA/Ro, SSB/La, U1RNP (70 kDa, A, C), Scl-70, JO-1, centromere B and Sm. Also, anti-dsDNA (EliA dsDNA, Pharmacia Diagnostics, Freiburg, Germany) was measured on all samples. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), efficiency, positive likelihood ratio (PLR), and negative likelihood ratio (NLR) were calculated. RESULTS: Diagnostic efficiency was similar for IIF (82.6%) and EliA Symphony (82.3%), as well as PLR (6.5 for IIF, and 7.3 for Eli Symphony), and NLR (0.35 for IIF, and 0.41 for EliA Symphony). The combined measurement of EliA Symphony and dsDNA increased sensitivity but not PLR. Area under receiver operator characteristic (ROC) curve was similar for IIF (0.847) and EliA Symphony (0.823). CONCLUSIONS: The results of the study demonstrate that EliA Symphony solely or combined with anti-dsDNA detection has an efficiency similar to HEp-2 cells IIF with a cut-off of 1:160 for the diagnosis of CTD. PMID- 15894302 TI - What does an intermediate success rate mean? An analysis of a Piagetian liquid conservation task in the great apes. AB - The study investigates what an intermediate success rate means in bonobos, chimpanzees, and orangutans. Apes participated in liquid conservation experiments where they had to track the larger of two different quantities of juice after various kinds of transformations [Suda, C., & Call, J. (2004). Piagetian liquid conservation in the great apes (Pan paniscus, Pan troglodytes, and Pongo pygmaeus). Journal of Comparative Psychology, 118, 265-279). When making a decision, apes sometimes demonstrated hesitant behavior, concurrently pointing to both alternatives or successively changing their choice. Moderately successful apes showed more hesitation than highly successful or unsuccessful apes. The results are consistent with the cognitive conflict model: The experiments created a higher degree of cognitive conflict on moderately successful apes than on very successful or unsuccessful apes. This indicates that an intermediate performance reflects the joint operation and potential conflict between two different cognitive strategies (identity and appearance) inherent to the Piagetian conservation task. PMID- 15894303 TI - Estradiol protects against alteration of protein kinase C(epsilon) in a binge model of ethanol dependence and withdrawal. AB - This study tested the hypothesis that a binge type of ethanol intake and ethanol withdrawal disturbs protein kinase C (PKC) homeostasis in a manner protected by 17beta-estradiol. Ovariectomized rats implanted with 17beta-estradiol or oil pellets received ethanol (7.5% weight/volume, 7 days) or control solution by a gavage method. The cerebelli were collected during ethanol exposure or ethanol withdrawal to assess the activity, protein levels, and cellular distribution of PKC(epsilon) and total PKC, using an ATP phosphorylation and immunoblot assays. While both ethanol exposure and ethanol withdrawal increased membrane protein levels and membrane translocation, only ethanol withdrawal enhanced activity of PKC(epsilon). Ethanol withdrawal not ethanol exposure increased the three parameters of total PKC. 17beta-Estradiol treatment prevented these changes in PKC profiles. These data suggest that an excessive episodic intake of ethanol followed by ethanol withdrawal disturbs PKC homeostasis and cellular distribution of PKC, in particular PKC(epsilon), in a manner that is protected by estrogen. PKC(epsilon) appears more vulnerable during ethanol withdrawal than during ethanol exposure. PMID- 15894304 TI - Time and brain region specific up-regulation of low affinity neuronal nicotinic receptors during chronic nicotine administration in mice. AB - We studied the effects of chronic oral nicotine on brain low affinity nicotine binding sites. Mice received nicotine in the drinking water for 4 or 7 weeks. Receptor binding was measured at 24 or 48 h after cessation of nicotine administration with [3H]methyllycaconitine, an antagonist in alpha7 and alpha3/alpha6beta2beta3* nicotinic receptors in striatum, midbrain, hippocampus and cortex. Chronic nicotine for 4 weeks resulted in a significant increase in the [3H]methyllycaconitine binding in the striatum and cortex, whereas after 7 weeks the increase in binding could be found in the hippocampus but not in the other brain areas studied. For comparison, high affinity nicotine binding sites (mostly alpha4beta2) were measured with [3H]epibatidine after 7-week chronic nicotine treatment. [3H]Epibatidine binding sites were increased in the hippocampus, midbrain and cortex, but not in the striatum. The up-regulation of [3H]methyllycaconitine binding was significant at 24 h but that of [3H]epibatidine binding sites was not observed until at 48 h after cessation of chronic nicotine. These results suggest that up-regulation of low affinity nicotine binding sites does occur during chronic nicotine administration. Furthermore, the low affinity and high affinity binding differ clearly as regards regions and duration suggesting that different nicotinic receptors respond differently to nicotine administration. PMID- 15894305 TI - The effect of gliclazide and glibenclamide on preconditioning of the human myocardium. AB - The cardioprotection of ischaemic preconditioning may be abolished in diabetic patients especially when some oral hypoglycaemics are used. The dose-response effect of gliclazide and glibenclamide on ischaemic preconditioning and the action of glibenclamide on signal transduction in human myocardium were investigated using right atrial appendages from cardiac surgery patients. Glibenclamide (0.1, 1, 3 and 10 microM) and gliclazide (1, 10, 30 and 100 microM) were added for 10 min prior to ischaemic preconditioning. The cardioprotection was abolished by glibenclamide at all concentrations and by gliclazide at supratherapeutic concentrations of 30 and 100 microM. Glibenclamide abolished the protective effect of mitoK(ATP) channel opening but not that of protein kinase C (PKC) or p38 mitogen activated protein kinase (p38MAPK) activation. In conclusion, glibenclamide and gliclazide differential effects may be a result of differential sensitivities. Glibenclamide does not block protection conferred by either PKC or p38MAPK activation. These findings may have clinical implications in ischaemic heart disease. PMID- 15894306 TI - Trapidil inhibits platelet-derived growth factor-induced migration via protein kinase A and RhoA/Rho-associated kinase in rat vascular smooth muscle cells. AB - Trapidil suppresses platelet-derived growth factor (PDGF)-induced vascular smooth muscle cell (VSMC) proliferation by inhibiting Raf-1/extracellular signal regulated kinase (ERK) via cAMP/protein kinase A (PKA). We examined whether trapidil inhibits PDGF-induced VSMC migration and investigated its mechanisms of action. VSMC migration was inhibited to a similar extent by trapidil and forskolin. A PKA inhibitor N-(2-[p-bromocinnamylamino]ethyl)-5 isoquinolinesulfonamide (H89) blocked the inhibition by forskolin to a greater degree than that by trapidil. Trapidil but not forskolin suppressed PDGF stimulated RhoA activation. In the presence of both H89 and (+)-(R)-trans-4-(1 aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide dihydrochloride monohydrate, an inhibitor of Rho-associated kinase (ROCK), trapidil and forskolin inhibited migration to a similar extent. Thus, in addition to cAMP/PKA activation, trapidil inhibits RhoA/ROCK activation, which may be important in trapidil's inhibitory effect on migration. PMID- 15894307 TI - Tacrolimus (FK506) has protective actions against murine bleomycin-induced acute lung injuries. AB - The effects of tacrolimus on murine acute lung injury were tested, especially in comparison to dexamethasone. Acute lung injury was induced by intratracheal instillation of bleomycin. Oral tacrolimus significantly improved survival rates of bleomycin-exposed mice, while cyclosporin A or dexamethasone did not. After instillation of bleomycin (day 0), a migration of neutrophils into alveolar spaces peaked on day 3, with concomitant increases of chemokines. On day 6, marked morphological changes in the lungs were observed. All these changes were significantly inhibited by tacrolimus. Furthermore, DNA ladder and immunohistochemical analyses of lungs showed that apoptosis of lung cells appeared on day 6 and was abolished only by the treatment of tacrolimus. These results suggest that both anti-inflammatory and anti-apoptotic action of tacrolimus contribute to improvement of bleomycin-induced acute lung injury. PMID- 15894308 TI - Endothelial mechanisms underlying responses to acetylcholine in the horse deep dorsal penile vein. AB - This study evaluates the mechanisms underlying endothelium-dependent responses to acetylcholine in horse deep dorsal penile veins. Acetylcholine-induced relaxation was abolished by endothelium removal, the soluble guanylyl cyclase-inhibitor, and the nitric oxide (NO) synthase inhibitors. Acetylcholine-induced relaxation was inhibited by high K+ concentrations and blockade of large-conductance Ca(2+) activated potassium (BK(Ca)) channels, and voltage-dependent potassium (K(v)) channels. Relaxations were unaffected by a small-conductance K(Ca) (SK(Ca)) channel blocker, or an ATP-sensitive potassium (K(ATP)) channel blocker. Relaxation in response to a NO donor was unaffected by K(Ca) channel blockers, but inhibited by high K+ concentrations and a K(v) channel blocker. In the presence of a NO synthase inhibitor, acetylcholine-induced contractions were inhibited by a cyclooxygenase blocker and abolished by endothelial removal. The contractile response was competitively inhibited by muscarinic receptor antagonists, high affinity M1 and M3 antagonists, while the M2 antagonist had no effect. The pharmacological profile suggests that acetylcholine contraction is mediated by muscarinic M1 receptors. Our findings indicate that acetylcholine induced relaxation in the horse deep dorsal penile vein is essentially mediated by NO, acting via the cGMP-dependent pathway and opening of K+ channels. The contraction elicited by acetylcholine is prostanoid-mediated and induced by endothelial muscarinic M1 receptor activation. PMID- 15894309 TI - Interaction of acetylcholine with Kir6.1 channels heterologously expressed in human embryonic kidney cells. AB - Kir6.1 subunit is one of the pore-forming components of K(ATP) channel complex. The endogenous modulation of Kir6.1 subunit function has been largely unknown. Whether acetylcholine modulated the function of Kir6.1 subunit stably expressed in human embryonic kidney (HEK-293) cells was examined in the present study using the whole-cell patch-clamp technique. Acetylcholine from 1-100 microM concentration-dependently stimulated the heteologously expressed and PNU-37883A sensitive Kir6.1 channels (p<0.05). Co-expression of sulphonylurea receptor 1 subunit with Kir6.1 significantly inhibited the stimulatory effect of acetylcholine on K(ATP) currents. Pretreatment of the transfected HEK-293 cells with atropine, alpha-bungarotoxin, mecamylamine, prazocine, propranolol, or dihydro-beta-erythroidine hydrobromide did not alter the stimulatory effect of acetylcholine on Kir6.1 currents. When intracellular ATP was increased from 0.3 mM to 5 mM, acetylcholine at 10 microM still exhibited its stimulatory effect ( 16.4+/-2.3 to -25.5+/-3.8 pA/pF, n=8, p<0.05). In conclusion, we have demonstrated an excitatory effect of acetylcholine on Kir6.1 channels, which is mediated neither by an acetylcholine receptor-dependent mechanism, nor by alteration in ATP metabolism. PMID- 15894310 TI - Synergistic interaction of gabapentin and oxcarbazepine in the mouse maximal electroshock seizure model--an isobolographic analysis. AB - The anticonvulsant effects produced by mixtures of oxcarbazepine and gabapentin (two second-generation antiepileptic drugs) in numerous fixed-ratio combinations of 1:1, 1:2, 1:5, 1:10, 1:15, and 1:20 were examined isobolographically in the mouse maximal electroshock seizure model. Results displayed that mixtures of both drugs at the fixed-ratios of 1:2, 1:5, 1:10, 1:15, and 1:20 exerted supra additive (synergistic) interactions against electroconvulsions. Only a fixed ratio of 1:1 was indifferent with isobolography, although the combination displayed the trend towards supra-additivity. Furthermore, the combinations of oxcarbazepine with gabapentin, administered at their median effective doses (ED(50 mix)s), did not alter motor performance of animals challenged with the chimney test. Additionally, neither gabapentin nor oxcarbazepine affected total brain concentrations of co-administered drug, indicating a pharmacodynamic nature of interaction between these antiepileptics. Finally, based on preclinical data presented here the combination of oxcarbazepine and gabapentin is of particular importance for further therapy in patients with refractory partial seizures. PMID- 15894311 TI - Aripiprazole's low intrinsic activities at human dopamine D2L and D2S receptors render it a unique antipsychotic. AB - Aripiprazole is the first clinically approved atypical antipsychotic agent having dopamine D2 receptor partial agonist activities. To evaluate aripiprazole's agonist and antagonist properties, we established a Chinese hamster ovary cell line expressing high and low densities of the long and short isoforms of human dopamine D2 receptors, then compared its properties with 7-{3-[4-(2,3 dimethylphenyl)piperazinyl]propoxy}-2(1H)-quinolinone (OPC-4392), S(-)-3-(3 hydroxyphenyl)-N-n-propylpiperidine ((-)-3-PPP), and terguride (other partial agonists) using forskolin-stimulated cAMP accumulation as an index. In cells expressing high receptor densities, all partial agonists predominantly behaved as agonists. However, in cells expressing low receptor densities, the partial agonists showed significantly lower maximal effects than dopamine. Aripiprazole showed the lowest intrinsic activities. In addition, all compounds blocked the action of dopamine with a maximum effect equal to that of each compound alone. Aripiprazole's low intrinsic activities may account for the clinical finding that, unlike the other partial agonists, it is substantially active against both positive and negative symptoms of schizophrenia. PMID- 15894312 TI - Quantitative multi-gene transcriptional profiling using real-time PCR with a master template. AB - We previously reported a method for quantitative multi-gene transcriptional profiling with gene-specific standard curves using real-time PCR. Here, we report an approach that increases experimental throughput by using a master template to generate a single standard curve for the estimation of mRNA copy numbers from all genes. We prepared fifty-nine different templates and measured eNOS mRNA copy numbers in Matrigel VEGF-transfectant samples. The copy numbers obtained using each of the fifty-nine templates were within 50% of the copy number obtained using the previously reported method. Analysis of primer design parameters, and subsequent tests, showed that eliminating complementarities between the first nucleotides at the 5'-ends of the forward and reverse primers reduces the influence of saturation effects and produces copy numbers similar to those generated with gene-specific templates-generally, within 20%. Measurements on a panel consisting of eNOS, iNOS, and nNOS further validated the master-template approach. The master-template approach enables rapid quantification of mRNA abundances from panels of hundreds of genes, and will be a valuable tool for screening large numbers of genes as part of a search for biomarkers, the validation of DNA-microarray data, or research into the dynamics of the gene protein network. PMID- 15894313 TI - Subconfluent endothelial cells form podosomes downstream of cytokine and RhoGTPase signaling. AB - Adhesion, migration and invasion of endothelial cells are prerequisites for the formation of blood vessels and have to be controlled on a subcellular level. We report that subconfluent human umbilical vein endothelial cells (HUVEC) are able to constitutively form podosomal adhesions that are sites of matrix metalloprotease concentration and matrix degradation. Importantly, incubation of serum-starved cells with VEGF or TNFalpha revealed the dependence of podosomes on cytokine signaling. Podosome formation was also stimulated by addition of monocytes to HUVEC. Microinjection/application of specific inhibitors or active/inactive mutants showed that regulatory pathways include Src kinase and RhoGTPase signaling, N-WASP activation and Arp2/3 complex-dependent actin nucleation. In sum, our data show that HUVEC displaying a migratory phenotype constitutively form f-actin-rich adhesions with podosomal characteristics downstream of cytokine signaling. We propose that HUVEC podosomes play an important role in endothelial cell migration and invasion. PMID- 15894314 TI - The mitochondrial ribosome-specific MrpL55 protein is essential in Drosophila and dynamically required during development. AB - We report on the essential Drosophila mRpL55 gene conserved exclusively in metazoans. Null mRpL55 mutants did not grow after hatching, moved slowly and died as first instar larvae. MrpL55 is similar to mammalian MRPL55, a protein that, in a large-scale mass spectrometry study, has been found as a mitoribosome-specific large subunit protein. We showed that MrpL55 was localised to the mitochondrion in S2 cells and tissues and was enriched in cells with a higher protein synthesis activity. The MrpL55 protein contains a KOW-like motif present in proteins with a role in transcriptional anti-termination and regulation of translation. Modulation of mRpL55 expression level is critical for development. Somatic clonal analysis showed that MrpL55 was not required in larval eye imaginal discs but required in pupal discs apparently during the second mitotic wave. Therefore, our results showed that the MrpL55 protein acts dynamically in the cell during development. We propose that MrpL55 is involved in Drosophila mitochondrial biogenesis and G2/M phase cell cycle progression. PMID- 15894315 TI - Laminins 2 (alpha2beta1gamma1, Lm-211) and 8 (alpha4beta1gamma1, Lm-411) are synthesized and secreted by tooth pulp fibroblasts and differentially promote neurite outgrowth from trigeminal ganglion sensory neurons. AB - The tooth pulp innervation originates from the trigeminal ganglion (TG) and represents an illustrative example of tissue targeting by sensory nerves. Pulpal fibroblasts strongly promote neurite outgrowth from TG neurons in vitro. In the present study, we have investigated the possible participation of laminins (LNs), potent neuritogenic extracellular matrix components. Immunohistochemistry of human tooth pulp demonstrated expression of LN alpha1, alpha2, alpha4, alpha5, beta1 and gamma1, and laminin-binding integrin alpha3, alpha6, beta1 and beta4 chains in nerves. Though faintly stained for laminins in situ, pulpal fibroblasts reacted, once cultured and permeabilized, with antibodies to LN alpha2, alpha4, beta1 and gamma1 chains by flow cytometry. The cells also expressed the corresponding mRNAs and were able to assemble and secrete LN-2 (alpha2beta1gamma1, Lm-211) and LN-8 (alpha4beta1gamma1, Lm-411). LN-8 displayed a chondroitin sulphate (CS) modification in its alpha4 chain. In functional assays, mouse LN-1 (alpha1beta1gamma1, Lm-111) and recombinant human (rh) LN-8, but not native or rhLN-2, strongly promoted neurite outgrowth from TG neurons, mimicking the effect of cultured pulp fibroblast. Altogether, the results indicate that LN-2 and LN-8 are synthesized by tooth pulp fibroblasts and differentially promote neurite outgrowth from TG neurons. LN-8 may contribute to sensory innervation of teeth and other tissues during development and/or regeneration. PMID- 15894316 TI - cDNA microarray reveals signaling pathways involved in hormones expression of human pituitary. AB - Pituitary, a master gland of neuroendocrine system, secretes hormones that orchestrate many physiological processes, under the regulation of multiple signaling pathways. To investigate the genes involved in hormones expression of human pituitary, homemade cDNA microarray containing 14,800 human genes/ESTs were used to profile the gene expression in both fetal and adult pituitaries. Seven hundred and twelve known genes changed over 2-fold between the both tissues. Of which, 23 genes were changed with hormones expression in aging were confirmed by RT-PCR, not only the known regulators such as Pit1, GATA4, ESRRA, GABA-A, and EMK, but also LOC55884, DUSP3, PNN, and RCL, which had not been reported to be involved in the hormones expression. Correspondingly, the mRNAs of GH, PRL, POMC, TSH-beta, FSH-beta, and LH-beta, was increased as much as 6- to 20-fold in adult pituitary than those in fetal pituitary, by real-time quantitative RT-PCR assay. In addition, the mRNAs of signaling pathways, such as cAMP-PKA-CREB, PI3K-Akt, and PKA-ERK were further investigated. Of them, it was only cAMP-PKA-CREB pathway, but not PI3K-Akt and PKA-ERK have the same expressing pattern as hormones. It suggested that cDNA microarray is highly advantages to profile the differential expressed genes that were involved in hormones expression of human pituitary, but it might ignore some responding proteins regulated posttranscriptionally. PMID- 15894317 TI - Purification of follicle-stimulating hormone from immature Japanese eel, Anguilla japonica, and its biochemical properties and steroidogenic activities. AB - Follicle-stimulating hormone (FSH) was purified, for the first time, from immature Japanese eel, Anguilla japonica, and its biochemical properties were investigated. FSH was extracted from immature eel pituitaries and purified by gel filtration on Sephadex G-100, and two step anion-exchange chromatography: stepwise elution on DE-52, followed by gradient elution on TSK-gel Super-Q using HPLC. Purification was performed using its molecular mass and the positive reaction with anti-Japanese eel (je) FSHbeta antiserum. Purified eel FSH was detected as a single band after separation by SDS-PAGE under a non-reducing condition, showing positive reaction with both anti-je glycoprotein (GP) alpha and anti-jeFSHbeta antisera. The molecular mass of purified eel FSH was estimated to be approximately 33 kDa. After separation by SDS-PAGE under reducing condition, the intact molecule was detected as distinct proteins, whose N terminus amino acid sequences coincided with those predicted from cDNA sequences for jeGPalpha and jeFSHbeta mature peptides. Deglycosylation of these subunits led to a decrease in their molecular mass. These results suggest that eel FSH is a heterodimeric molecule which consists of distinct glycoprotein subunits, GPalpha and FSHbeta. Cells reacting with anti-jeFSHbeta antiserum were observed in the proximal pars distalis of an immature eel pituitary, while jeLHbeta immunoreactive cells were not detected. Gonadotropic activities of eel FSH were demonstrated in vitro by stimulating testosterone and 11-ketotestosterone secretions in immature eel testes. Purified eel FSH stimulated the secretion of both androgens from the immature eel testis in a dose-dependent manner, similar to immature eel pituitary homogenate and recombinant eel FSH produced by yeast. These results show that endogenous and recombinant FSH in this species possess similar activities, presumably stimulating the gametogenesis through the sex steroid secretion during the early stages of gonadal development. PMID- 15894318 TI - Chronic stress reduces body fat content in both obesity-prone and obesity resistant strains of mice. AB - Unpredictable stressors have been used to assess the effect of stress on energy metabolism in obesity-prone (C57BL6J) and obesity-resistant (AJ) mice. Mice were exposed for 25 days to a stress protocol. Both strains of mice were divided into groups of control and stressed mice, which had access to either a high-fat or a high-carbohydrate diet. Twenty-four hours after the last session of stress, mice were sacrificed for blood and brain collections. Insulin, corticosterone, and glucose concentrations in plasma were measured, and expressions of corticotropin releasing factor (CRF) in the paraventricular hypothalamic nucleus (PVH) and the central amygdala (CeA) were determined by in situ hybridization. Stressed mice in all groups had lower body fat contents than control mice, and all mice fed with the high-fat diet had heavier retroperitoneal and inguinal fat pads than mice fed with carbohydrate. CRF mRNA level in the CeA was lower in B6 mice than in AJ mice. Stressed mice had a lower expression of CRF in the CeA than control mice. In conclusion, chronic stress reduces body fat content in obesity-prone as well as in obesity-resistant mice. PMID- 15894319 TI - Native pentameric C-reactive protein displays more potent pro-atherogenic activities in human aortic endothelial cells than modified C-reactive protein. AB - Inflammation is pivotal in atherogenesis. Numerous prospective studies have shown that levels of high sensitive C-reactive protein (CRP) predict cardiovascular events. Recently, data suggests that CRP could be a mediator in atherothrombosis. Loss of pentameric symmetry in CRP has been shown to result in the formation of modified CRP (mCRP). The main aim of this study was to examine the biological effects of the native, pentameric form of CRP compared to a modified form in human aortic endothelial cells. Human pentameric native CRP (n-CRP) from two different sources (recombinant and serum) was purified and used. It was then subjected to EDTA chelation and urea treatment to prepare modified CRP (m-CRP). Purity of both n-CRP and m-CRP preparations was checked by gel electrophoresis. Both n-CRP and m-CRP were incubated with human aortic endothelial cells (HAEC) and biological activities was tested by assaying for interleukin-8 (IL-8), plasminogen activator inhibitor-1(PAI-1), cyclic GMP and prostaglandin F1-alpha. n-CRP significantly upregulated IL-8 at concentrations > or = 10 microg/mL while m-CRP upregulated IL-8 only at concentrations of 50 microg/mL (p < 0.05). PAI-1 levels were significantly increased to a greater extent with native compared to m CRP (p < 0.05). While both decreased PGF1-alpha at concentrations of 50 microg/mL, the effect of native CRP was more pronounced and was evident at 10 microg/mL (p < 0.05). The most pronounced difference was observed with regard to inhibition of eNOS activity as assessed by cGMP which was observed at 10 microg/ml of native CRP but only at 50 microg/mL for m-CRP (native CRP versus mCRP: p < 0.001). Thus, native pentameric CRP compared to modified CRP exerts more potent atherogenic effects in human aortic endothelial cells. PMID- 15894320 TI - Circulating levels of MCP-1 and eotaxin are not associated with presence of atherosclerosis or previous myocardial infarction. AB - The chemokines are a family of signalling proteins that participate in regulation of the immune system and have been implicated in the pathogenesis of vascular diseases. Deleting the gene encoding the chemokine MCP-1 in mouse models of atherosclerosis reduces lipid lesion formation and circulating chemokines are upregulated in man immediately following myocardial infarction (MI) or coronary angioplasty. We have therefore investigated whether circulating levels of two chemokines (MCP-1 and eotaxin) differ between subjects with and without atherosclerosis. We have used three different methods of measuring the presence and extent of atherosclerosis in human subjects: duplex ultrasonography of the carotid arteries and clinical diagnosis of coronary heart disease on individuals from the general population and coronary angiography on patients with suspected heart disease. There was no difference in the levels of circulating MCP-1 or eotaxin, measured by ELISA, between subjects with and without atherosclerosis. Furthermore, any increase in circulating MCP-1 following acute MI must be short lived, since chemokine levels were not different in subjects who had had an MI previously compared to those who had not. We conclude that although there may be a transient increase in circulating chemokine levels following coronary angioplasty, there is no difference in the levels of circulating MCP-1 or eotaxin in subjects with and without atherosclerosis. PMID- 15894321 TI - Fibrates and coronary risk reduction. AB - Many medications are currently available to correct lipoprotein abnormalities when lifestyle measures alone are not sufficient. No single agent or class of agents is able to correct all of the lipoprotein abnormalities. This paper reviews the role of one class, the fibrates, in the management of lipid disorders and summarizes the clinical trial information relating to their impact on coronary artery disease. PMID- 15894322 TI - Problematic sites of third body embedment in polyethylene for total hip wear acceleration. AB - A computational model was developed to identify the sites of third body particle embedment in a total hip acetabular component surface that are most problematic in terms of roughening the overpassing regions of the femoral head counterface, leading in turn to most severely accelerated polyethylene wear. The analytical approach used was to calculate loci of acetabular sites that, during the gait cycle, overpass previously documented regions of kinetically most critical femoral head roughening. Instantaneous local contact stress and sliding distance were postulated as factors contributing to the severity of the femoral head scratching/roughening which would be expected, due to otherwise-similar particles embedded along each such acetabular overpass locus. The computational results showed that the location of debris embedment was a potent determinant of the amount of polyethylene wear acceleration expected. The data also showed that the supero-lateral aspect of the acetabular cup is consistently and by far the most problematic area for third body particle embedment. PMID- 15894323 TI - Human calf muscle responses during repeated isometric plantarflexions. AB - In the present study, we measured the contraction-induced shortening (dL) of individual synergistic human muscles in a repeated motor task to assess their contractile behaviour. Ultrasonography was used to obtain dL measurements in the gastrocnemius (GS) and soleus (SOL) muscles of six men performing 11 consecutive isometric plantarflexions. Contractions 1 and 11 were performed with maximal effort, and contractions 2-4, 5-7 and 8-10 were performed with efforts generating 50, 70 and 90%, respectively, of the plantarflexion moment produced in contraction 1. In contractions 5-10, the SOL muscle dL was similar (p > 0.05) to that produced in contraction 1 (approximately 6 mm), indicating that the SOL muscle became fully activated at 70% of the maximum plantarflexion moment. The GS muscle dL in contractions 10 and 11 exceeded by approximately 0.5 mm (p < 0.05) and 1.3 mm (p < 0.01), respectively, that generated in contraction 1 (approximately 10 mm), despite evidence obtained by superimposed stimulation that contraction 1 was produced with full motor unit activation. The consequent paradox that the GS muscle would produce in contractions 10 and 11 a greater activation and therefore more force than its actual potential is resolved when considering the interaction between the time-dependent tensile response of tendon and the performance of muscle as dictated by the sliding filament mechanism of contraction. PMID- 15894324 TI - The safe-zones for combined cup and neck anteversions that fulfill the essential range of motion and their optimum combination in total hip replacements. AB - Reduction of the range of motion (ROM) until prosthetic impingement of a total hip replacement may lead to frequent impingement, subluxation and dislocation especially for patients with good hip movement. The ROM until prosthetic impingement can be calculated using the technical ROM (theta) and the cup and neck positions by a previously created mathematical formula. A larger (theta) with proper cup and neck positions results in a larger ROM. However there was only one paper written in English, which revealed the optimum theoretical combination of cup and neck anteversions. ROM of more than 110 degrees flexion, 30 degrees internal-rotation at 90 degrees flexion, 30 degrees extension and 40 degrees external-rotation were defined as the criteria for essential ROM for ADL. The safe-zones for combined cup anteversion (betaanat) and neck anteversion (b) were defined as the areas that fulfill all the criteria of ROM without prosthetic impingement. The safe-zones were created for 35 degrees , 45 degrees and 55 degrees cup abductions (alpha) and for 120 degrees and 135 degrees (theta). The safe-zones for combined (betaanat) and (b) were much larger for a 135 degrees (theta) than a 120 degrees (theta). Their safe-zones showed that (b) should be reduced if (betaanat) is increased and choosing a lower (alpha) requires that the sum of (betaanat) and (b) should be higher and vice versa. A (theta) of more than 135 degrees is recommended as it further increases the size of the safe-zone and provides a larger ROM, and the optimum values of combined cup and neck anteversions can be estimated by the formula: (alpha) + (betaanat) + 0.77(b) = 84.3. PMID- 15894325 TI - Simulation of Codman's paradox reveals a general law of motion. AB - Codman's paradox refers to a specific pattern of motion at the shoulder joint. It asked how a mysterious axial rotation about the longitudinal axis of the arm occurred during two or three sequential arm rotations that did not involve rotation about the long-axis. The objective of this paper was to find how the mysterious axial rotation occurred in the Codman's paradox. First, Codman's paradox and Codman's rotation were defined in general situations that involved arm rotations about orthogonal axes starting from the neutral attitude as well as rotations about non-orthogonal axes and starting from non-neutral attitudes. Then a general law of motion was proposed to answer the question of the Codman's paradox, which is stated as when the long-axis of the arm performs a closed-loop motion by three sequential rotations defined as Codman's rotation, it produces an equivalent axial rotation angle about the long-axis. The equivalent axial rotation angle equals the angle of swing-the second rotation in the three sequential long-axis rotations. Validity of the proposed law of motion is demonstrated by computer simulation of various Codman's rotations. Clinical relevance of the proposed law of motion is also discussed in the paper. PMID- 15894326 TI - Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV). AB - Accurate assessment of neutralizing antibody activities is important either for patients infected with Severe Acute Respiratory Syndrome (SARS) or for animals and volunteers immunized with the experimental vaccines against the SARS associated coronavirus (SCV). However, the current assay based on the cytopathic effect (CPE) which has been frequently cited in literature has several limitations. The CPE assay relies on the visual observation on the damage of SCV infected target cells under a microscope. It is subjected to observer variations and it is difficult to generate a quantitative determination of neutralizing activities based on the level of CPE. In the current study, we established the utility of two additional assays to measure the neutralizing activities against SCV: the plaque reduction (PR) and the neutral red staining (NRS) assays. The PR assay described in this study was modified from the traditional viral plaque reduction assay by using an improved crystal staining method to achieve better plague formation in SCV infected Vero E6 cells. The NRS neutralization assay was adopted from a similar system used for detecting neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). In this assay, the protective effect of neutralizing antibodies was determined by the cell viability which is measured by the uptake of neutral red dye at A540. The neutralizing antibody titers can be easily determined with either of the two new assays. In this report, we described the utility of these two new neutralization assays in measuring the neutralizing activities against SCV infection from rabbit sera immunized with various forms of spike protein of SCV. PMID- 15894327 TI - The human Burkitt lymphoma cell line Namalwa represents a homogenous cell system characterized by high levels of Toll-like receptor 9 and activation by CpG oligonucleotides. AB - Human B cells and plasmacytoid dendritic cells constitutively express Toll-like receptor (TLR)9 and respond to TLR9 ligands, as evidenced by nuclear factor kappa B translocation and cytokine secretion. However, TLR9 expression on B lymphocytes appears to be dependent upon both the state of activation and differentiation of the B cell population. In the current study, TLR9 mRNA expression was evaluated in transformed human B cell lines and correlated with their response to CpG. Among the B cell lines, the Burkitt lymphoma-derived Namalwa line had the highest level of TLR9 expression, 20-fold greater than spleen. Following incubation with CpG oligonucleotide, Namalwa cells secreted increased amounts of TNF, IL-6, and IL-10 and expressed the costimulator molecules CD40, CD80, and CD86. These functional responses to TLR9 activation occurred with similar EC(50) values in the 30- to 60-nM range. These results suggest that the Namalwa Burkitt lymphoma line may serve as a useful cell-based assay for the detection of novel TLR9 agonists as well as a model to further explore the regulation of TLR9 expression and signaling. PMID- 15894328 TI - Measurement of vasoactive intestinal peptide using a competitive fluorescent microsphere immunoassay or ELISA in human blood samples. AB - The concentration of Vasoactive Intestinal Peptide (VIP) as measured by recycling immunoaffinity chromatography (RIC) has been reported to be elevated in the blood of patients with autism as compared with normal subjects. In this study, we have developed a "Competitive Fluorescent Microsphere Immunoassay" (cFMI) in which VIP competes with biotinylated VIP in binding to polyclonal antibodies on microspheres. The results were obtained using the Luminex100 system. We measured VIP in serum, plasma, and material eluted from dried blood spots on filter paper with both the cFMI and an ELISA procedure. We found that a purification procedure was necessary for obtaining useful results from plasma and serum, however, a preincubation step was required with the blood eluates. This newly developed cFMI was more sensitive (2.5 vs. 20.0 pg/ml), and more reproducible than the ELISA. To get accurate measurements of VIP in eluted material high sensitivity is especially important. Thus, the cFMI using the Luminex system has definite advantages over a conventional ELISA including the possibility that samples can be assayed at higher dilutions. We have determined that the VIP concentrations of serum, plasma, and dried blood spot eluate specimens as measured with the cFMI assay system were similar to those measured with ELISA. Thus, the new cFMI using Luminex system may be useful for detection of VIP in human blood samples. PMID- 15894329 TI - The nature of Thelohania solenopsae (Microsporidia) cysts in abdomens of red imported fire ants, Solenopsis invicta. AB - Sixty four percent of Solenopsis invicta workers infected with Thelohania solenopsis contained 1-6 "cysts" ranging from 70 to 260 microm in diameter. Light and electron microscope analyses showed that cysts are hypertrophied adipocytes transformed by the parasites, each cyst presumably forming from a single cell. In the first step of the pathogenesis, Nosema-like spores functioning in autoinfection are produced; a diplokaryotic sequence leading to their formation causes fat body hypertrophy. When meiosis occurs, it switches parasite development to production of octospores and/or megaspores. Adipocytes become 2 4xlarger than normal in conjunction with intensive parasite multiplication and octospore maturation. Infected cells eventually lose their cellular organization and are converted into reservoirs for spores. There were no manifestations of cellular immunity, such as encapsulation or nodule formation. Similarly, there were no signs of specialized host-parasite interaction that might be interpreted as xenoma-like complexes. The role of the cysts in the parasite's life cycle is unclear. They may represent a defensive reaction of the host sacrificing the infected cells to segregate the infection. Alternatively, the cyst may help protect spores from environmental hazards and provide a concentrated infectious dose to aid horizontal transmission of the microsporidium. We propose to refer to hypertrophied adipocytes filled with T. solenospsae spores as "sporocytosacs", not "cysts." PMID- 15894330 TI - The remarkable mechanical strength of polycystin-1 supports a direct role in mechanotransduction. AB - Polycystin-1 is a large membrane-associated protein that interacts with polycystin-2 in the primary cilia of renal epithelial cells to form a mechanosensitive ion channel. Bending of the cilia induces calcium flow into the cells, mediated by the polycystin complex. Antibodies to polycystin-1 and polycystin-2 abolish this activation. Based on this, it has been suggested that the extracellular region of polycystin-1, which has a number of putative binding domains, may act as a mechanosensor. A large proportion of the extracellular region of polycystin-1 consists of beta-sandwich PKD domains in tandem array. We use atomic force microscopy to investigate the mechanical properties of the PKD domains of polycystin-1. We show that these domains, despite having a low thermodynamic stability, exhibit a remarkable mechanical strength, similar to that of immunoglobulin domains in the giant muscle protein titin. In agreement with the experimental results molecular dynamics simulations performed at low constant force show that the first PKD domain of polycystin (PKDd1) has a similar unfolding time as titin I27, under the same conditions. The simulations suggest that the basis for this mechanical stability is the formation of a force stabilised intermediate. Our results suggest that these domains will remain folded under external force supporting the hypothesis that polycystin-1 could act as a mechanosensor, detecting changes in fluid flow in the kidney tubule. PMID- 15894331 TI - Cannabinoids and neuroprotection in CNS inflammatory disease. AB - The current failure of potent immunosuppressive agents to control progressive disease in multiple sclerosis has moved a focus from immunotherapy towards the need for neuroprotection. There is increasing evidence for cannabinoid-mediated control of symptoms, which is being more supported by the underlying biology. However there is accumulating evidence in vitro and in vivo to support the hypothesis that the cannabinoid system can limit the neurodegenerative possesses that drive progressive disease, and may provide a new avenue for disease control. PMID- 15894332 TI - Genetic dissection of neurodegeneration and CNS inflammation. AB - Inflammation and neurodegeneration characterize multiple sclerosis, as well as many other diseases of the central nervous system (CNS). The understanding of the molecular pathways that regulate these processes is of fundamental importance for the development of new therapies. Nerve lesions paradigms in animals can serve as important tools to dissect central features of human CNS disease and by using these models certain key regulators have also been identified. However, our knowledge of how aspects of neurodegeneration and CNS inflammation are regulated on a genomic level is very limited. Such knowledge may help to unravel disease mechanisms. By using a standardized nerve trauma model, ventral root avulsion (VRA), in a series of inbred rat strains we here demonstrate a potent genetic regulation of the degree of neuron death and glial activation. Genome wide mapping of these phenotypes in experimental rat strain crosses identifies several quantitative trait loci (QTLs) controlling nerve lesion-induced nerve cell death, local T cell accumulation and expression of MHC class II on microglia. This approach may lead to the identification of evolutionary conserved genetic polymorphisms in key controlling genes, which can serve as prime candidates for association studies in several human CNS diseases. PMID- 15894333 TI - Axonal signals in central nervous system myelination, demyelination and remyelination. AB - Axonal signals are key players in central nervous system myelination. During development, the onset of myelination depends on a balance between positive and negative axonal signals. Among negative signals are inhibitory adhesion molecules that need to be removed from the cell surface for the myelination process to proceed. Positive signals necessary to initiate myelination consist of both interactions with specific adhesion molecules and electrical activity-induced release of promyelinating factors. In multiple sclerosis, demyelination induces major modifications of axonal surface components. The disruption of these factors might participate to the failure of the myelin repair process. PMID- 15894334 TI - Sodium-mediated axonal degeneration in inflammatory demyelinating disease. AB - Axonal degeneration is a major cause of permanent neurological deficit in multiple sclerosis (MS). The mechanisms responsible for the degeneration remain unclear, but evidence suggests that a failure to maintain axonal sodium ion homeostasis may be a key step that underlies at least some of the degeneration. Sodium ions can accumulate within axons due to a series of events, including impulse activity and exposure to inflammatory factors such as nitric oxide. Recent findings have demonstrated that partial blockade of sodium channels can protect axons from nitric oxide-mediated degeneration in vitro, and from the effects of neuroinflammatory disease in vivo. This review describes some of the reasons why sodium ions might be expected to accumulate within axons in MS, and recent observations suggesting that it is possible to protect axons from degeneration in neuroinflammatory disease by partial sodium channel blockade. PMID- 15894335 TI - Neuroprotection by IL-10-producing MOG CD4+ T cells following ischemic stroke. AB - Mucosal tolerance has been used successfully to treat animal models of autoimmune diseases and is being tested in human diseases. In this work we demonstrate the reduction of infarct size following mucosal tolerance by myelin oligodendrocyte glycoprotein (MOG) (35-55) peptide in mouse stroke model. Nasal MOG was most efficacious and reduced ischemic infarct size by 70% at 24 h as well as improving behavior score. Using immunohistological methods and IL-10 -/- mice, we demonstrate the importance of IL-10-producing CD4+ T cells in the reduction of the ischemic infarct volume following middle cerebral artery occlusion (MCAO). Furthermore, adoptive transfer of CD4+ T cells from nasally tolerized mice to untreated mice prior to MCAO surgery significantly decreased stroke size (p<0.001 vs. control), whereas CD4+ T cells from nasally tolerized IL-10-deficient mice had no significant effect. Based on these results, modulation of cerebral inflammation by mucosal tolerance to myelin antigens may have applicability both as prophylactic therapy and treatment following ischemia attacks. PMID- 15894336 TI - Role of cGMP signals in tetramethylpyrazine induced relaxation of the isolated rat aortic strip. AB - In the present study, role of guanosine-3',5'-cyclic monophosphate (cGMP) in the vasodilatation of tetramethylpyrazine (TMP), one of the active ingredients of the Chinese herb Chuang-xion, was investigated. We found that the TMP could decrease the vascular tone of isolated rat aorta precontracted with phenylephrine (10(-8) M) in a concentration-dependent manner from 10(-5) M to 10(-3) M. Also, the TMP induced relaxation was reduced by 1H-(1,2,4)-oxadiazol-(4,3-a)-quinoxalin-1-one (ODQ) or methylene blue, the inhibitor of soluble guanylyl cyclase. Moreover, the vasodilative response to TMP was enhanced significantly in the presence of sildenafil, a well-known inhibitor of phosphodiestrase type 5 that is sensitive to cGMP. In addition, TMP could increase the cGMP level in the isolated aortic rings and TMP-induced vasodilatation was deleted by cGMP-dependent protein kinases (PKG) blockade. These results suggest that relaxation of rat aortic strip by TMP is induced in the cGMP-dependent manner. PMID- 15894337 TI - Different pharmacological profile of two closely related endocannabinoid ester analogs. AB - The pharmacological and neuroprotective properties of two ester analogs of the endocannabinoids, arachidonoylethyleneglycol (AA-EG) and alpha,alpha,-dimethyl arachidonoylethyleneglycol (DMA-EG), were investigated. We examined the interaction of both compounds with cannabinoid receptors (CB1 and CB2) and their efficacy in functional assays. In competition binding assays, AA-EG and DMA-EG had low potency to displace the CB1/CB2 agonist [3H]CP-55,940 in membrane preparations expressing rodent or human receptors. Binding data correlate with low efficacy of both compounds as regards to inhibition of adenylyl cyclase activity. It was also shown that DMA-EG resists hydrolysis by rat brain membranes while AA-EG undergo complete splitting under these conditions. In the cannabinoid tetrad, AA-EG induced hypomotility, analgesia, catalepsy and decreased rectal temperature indicating cannabimimetic activity. By contrast, DMA-EG was completely inactive in the same models. DMA-EG and AA-EG potently protected rat cortical neurons in culture against oxygen deprivation at nanomolar concentrations. In glutamate-induced damage, the compounds were less active protecting neurons at micromolar concentrations. The data obtained indicate that the ester endocannabinoid template can be used for the development of new compounds with potent biological activity lacking some of the undesirable behavioral side effects. PMID- 15894338 TI - alpha-Adrenergic and muscarinic cholinergic receptors are not involved in the modulation of the parasympathetic baroreflex by the medial prefrontal cortex in rats. AB - The medial prefrontal cortex (MPFC) is involved in cardiovascular control and baroreflex modulation. Recent studies indicated that stimulation of MPFC muscarinic receptors causes hypotensive responses whereas stimulation of alpha1- but not of alpha2-adrenoceptors causes pressor responses in unanesthetized rats. It has also been shown that the MPFC is involved in the modulation of the parasympathetic component of the baroreflex in rats. We report that bilateral injections of CoCl2 in the ventral portion of the MPFC (vMPFC) reduced the parasympathetic component of the baroreflex, thus confirming the involvement of local synapses. We further evaluated the effect of the pharmacologic block of vMPFC alpha1- or alpha2-adrenoceptors and muscarinic receptors on the vMPFC related modulation of the parasympathetic component of the baroreflex in unanesthetized rats. Bilateral microinjections of 10 nmol of the selective alpha1 adrenoceptor antagonist WB4101 or 10 nmol of the selective alpha2-adrenoceptors antagonist RX821002 into the MPFC did not affect the baroreflex. Bilateral microinjections of 9 nmol of the muscarinic antagonist atropine also did not affect baroreflex activity. The present results indicate that although vMPFC alpha-adrenergic and muscarinic receptors are involved in cardiovascular regulation, they do not mediate the vMPFC-related modulation of the parasympathetic component of the baroreflex. PMID- 15894339 TI - Stimulation of acyl-CoA oxidase by alpha-linolenic acid-rich perilla oil lowers plasma triacylglycerol level in rats. AB - The effect of dietary polyunsaturated fatty acids (PUFA) on hepatic peroxisomal oxidation was investigated with respect to the postprandial triacylglycerol levels. Male Sprague--Dawley rats were fed semipurified diets containing either 1% (w/w) corn oil, or 10% each of beef tallow, corn oil, perilla oil, and fish oil for 4 weeks and 4 days. Hepatic and plasma triacylglycerol levels were reduced in rats fed fish and perilla oil diets compared with corn oil and beef tallow diets. The peroxisomal beta-oxidation, catalase activity, and acyl-CoA oxidase (AOX) activity were markedly increased by fish oil feeding. To a lesser extent, perilla oil elevated AOX activity in a 4-day feeding although the effect gradually decreased in a 4-week feeding. Similarly, the mRNA levels were increased in rats fed fish and perilla oils. AOX activity was negatively correlated with postprandial triacylglycerol levels. In addition, the stimulation of AOX was highly associated with the content of long chain n-3 PUFA such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in hepatic microsome. These effects were evident within 4 days of initiating feeding. Therefore, alpha linolenic perilla oil exerts a similar effect to fish oil in stimulating hepatic activity and gene expression of AOX by enriching long chain n-3 PUFA in hepatic membrane fraction, which can partly account for the reduction of postprandial triglyceridemia. PMID- 15894340 TI - Nature, distribution and origin of polycyclic aromatic hydrocarbons (PAHs) in the sediments of Olbia harbor (Northern Sardinia, Italy). AB - The nature, origin and distribution of US EPA polycyclic aromatic hydrocarbons (PAHs) in the sediments of Olbia harbor (North Sardinia, Italy) were investigated by gas-chromatography/mass spectrometry (GCMS). PAH concentrations in the sediments (SigmaPAHs) ranged from 0.16 to 0.77 microg g(-1), indicating a homogeneously low level of pollution. A rather exceptional prevalence of low molecular weight PAHs was substantiated: nearly 80% of SigmaPAHs include naphthalene (15.19%) and phenanthrene (64.47%). Carcinogenic compounds were present in very low (BaP, BkF, BaA and DBA) or negligible (BbF and Inp) concentrations. As indicated by the Phen/Ant molar ratio, the main source of PAHs is petrogenic, probably due to oil spills from shipping. The low/high molecular weight ratio (ranging between 1.2 and 26) distinguishes the sediments of the tourist harbor from those of the commercial/industrial harbor. Moreover, a slight but meaningful pyrolytic contribution to pollution was found in the tourist harbor, which has the most polluted sediments in the whole harbor. Finally, good linear correlations were found between a selected PAH (Phen and Naph) and selected sums of PAHs (i.e. the total amount of the other 15 US EPA and the sum of low molecular weight PAHs). PMID- 15894341 TI - Imposex levels in the dogwhelk Nucella lapillus (L.)--continuing improvement at high latitudes. AB - In 1990, restrictions on the use of tributyltin (TBT)-based antifouling paints were implemented in Iceland. A previous study showed that the level of imposex in the dogwhelk, Nucella lapillus, in Icelandic waters had decreased significantly between 1992 and 1998. In this study, we repeated the survey on imposex in N. lapillus at 33 locations from the Icelandic coast in 2003. The results indicated that both Vas Deferens Sequence Index (VDSI) and Relative Penis Size Index (RPSI) had further declined in 13 locations since 1998. Among these 13 sites, RPSI was reduced to zero in five cases. While improvements from 1992/1993 to 1998 were seen in reduced levels of imposex near both large and small harbours, the pattern from 1998 to 2003 was somewhat different, with improvement mainly observed near smaller harbours. No significant changes in imposex levels near larger harbours occurred over this period. Although the imposex levels still remain high near the large harbour complexes in Reykjavik and Hafnarfjordur, it is evident that regulations, including the use of less toxic antifouling paints and community action, have lead to substantial improvements in the marine environment of Iceland. International Maritime Organisation's ban on the application of TBT after 2003 is apparently necessary to allow further improvements in larger harbours. The environmental effects of new antifoulants replacing TBT need to be further evaluated. PMID- 15894342 TI - Heterogeneity of capillary flow in the retrograde microcirculation induced in rat limb by arteriovenous shunting. AB - Arteriovenous (AV) fistulas have been used clinically for improving adjunctive bypass patency. Such AV shunting induces retrograde flow in the microvascular network, which may induce microvascular remodeling and angiogenesis at the chronic phase. This paper was aimed to examine heterogeneity of blood flow among capillaries in the retrograde microcirculation induced by AV shunting. An AV anastomosis was created in rat hind limb. Using a dual window method or frame-by frame technique on the fluorescence microscopic video images, we measured velocities of red blood cells (RBCs) flowing in the capillary network in three flow conditions: control (normal flow), arterial occlusion, and AV shunting (retrograde flow). For each flow condition, RBC velocities were obtained in 155 capillaries of 6 rats. By classifying all the capillaries into four groups based on the levels of RBC velocity in the occlusion state, we evaluated the mean velocities, coefficient of variation (CV), and histograms for each group of capillaries. The mean velocity and CV in each group changed significantly from the control to AV shunting states. Especially, most significant changes appeared in capillary groups where the superficial femoral artery or its collateral arteries might have a direct influence. Though the AV shunting improved capillary perfusion in the mean level, major parts of capillaries still remained at low perfusion. PMID- 15894343 TI - Environmental enrichment alters plus maze, but not maternal defense performance in mice. AB - Maternal defense behavior (or maternal aggression) is a highly conserved behavior for protecting offspring that is normally associated with decreased fear and anxiety in rodent dams. Environmental enrichment can decrease indices of fearfulness and anxiety (and elevate novelty seeking) in laboratory animals. This study examined whether enrichment could alter levels of maternal aggression and elevated plus maze performance in lactating mice. One group of female mice was exposed to a series of novel objects for 1.5 months while the other group was not. As expected, mice that had been exposed to an enriched environment showed significantly more entries to and time on the open arms of an elevated plus maze while they were lactating. Enriched mice showed similar maze performances when tested 3 months later in a non-lactating state, even though enrichment had been removed for 3 months. Further, offspring of enriched dams exhibited maze performance similar to that of their dams, suggesting possible epigenetic influences on maze performance. In contrast, no differences in measures of maternal aggression or pup retrieval were detected between enriched and control lactating mice. Together, these results indicate that environmental enrichment has long lasting effects on plus maze performance, but does not alter maternal aggression. The findings also suggest that whatever neural pathways are altered by enrichment do not strongly overlap or regulate those governing maternal aggression. PMID- 15894344 TI - Heart rate variability in dairy cows-influences of breed and milking system. AB - Heart rate variability parameters in the time, frequency and nonlinear domains were investigated in two breeds of dairy cows (Austrian Simmental and Brown Swiss) milked either in an automatic milking system with partially forced cow traffic or in a herringbone milking parlour. Recordings were made of 24 cows (six of each breed and milking system) during lying, standing idle, and standing being milked, and analysed with linear mixed effects models taking the covariates time of day, live body weight, milk yield, stage of lactation and stage of pregnancy into account. Heart rate and nonlinear deterministic shares were higher, and heart rate variability in the time and frequency domains was lower, later in the day, in cows with higher body weight and in Simmental compared to Brown Swiss cows. Differences in the linear and nonlinear domains during lying indicated an increased level of chronic stress in cows in the automatic milking system with partially forced cow traffic, compared to cows milked in the herringbone milking parlour. No effects of milking system were found during milking, indicating that the stressor in the automatic milking system was not the milking process itself. PMID- 15894345 TI - Health system frailties in tuberculosis service provision in Russia: an analysis through the lens of formal nutritional support. AB - OBJECTIVE: The aim of this study was to describe health system challenges faced by tuberculosis (TB) treatment facilities in Russia through an analysis of formal institutional dietary provisions to patients in an inpatient facility that provides care for poor patients. METHODS: Analysis of formal dietary provisions by institutions and financing data from TB hospitals in Samara Oblast, Russia. RESULTS: Formal dietary provision for inpatients with TB has fallen substantially in recent years. In a hospital providing inpatient care for the poorest patients with fewest social support networks, this has been very pronounced. The likely reason for this is that financial support for other budget lines, principally salaries, has required protection. CONCLUSION: Formal institutional nutritional support in institutions providing care for the poorest patients with TB is unlikely to be enhancing the speed of recovery, or reducing the duration of infectiousness. Furthermore, the role that hospital may have played in the past in enabling patients to regain weight lost before admission may have been limited by reductions in formal financing. Reductions in state provision of food for patients may serve as an important illustration of wider TB control system frailties in the Russian Federation. PMID- 15894346 TI - Initial ocular following in humans: a response to first-order motion energy. AB - Visual motion is sensed by low-level (energy-based) and high-level (feature based) mechanisms. Ocular following responses (OFR) were elicited in humans by applying horizontal motion to vertical square-wave gratings lacking the fundamental ("missing fundamental stimulus"). Motion consisted of successive 1/4 wavelength steps, so the features and 4n+1 harmonics (where n=integer) shifted forwards, whereas the 4n-1 harmonics--including the strongest Fourier component (the 3rd harmonic)--shifted backwards (spatial aliasing). Initial OFR, recorded with the electromagnetic search coil technique, were always in the direction of the 3rd harmonic, e.g., leftward steps resulted in rightward OFR. Thus, the earliest OFR were strongly dependent on the motion of the major Fourier component, consistent with early spatio-temporal filtering prior to motion detection, as in the well-known energy model of motion analysis. PMID- 15894347 TI - The role of (micro)saccades and blinks in perceptual bi-stability from slant rivalry. AB - We exposed the visual system to an ambiguous 3D slant rivalry stimulus consisting of a grid for which monocular (perspective) and binocular (disparity) cues independently specified a slant about a horizontal axis. When those cues specified similar slants, observers perceived a single slant. When the difference between the specified slants was large, observers alternatively perceived a perspective- or a disparity-dominated slant. Eye movement measurements revealed that there was no positive correlation between a perceptual flip and both saccades (microsaccades as well as larger saccades) and blinks that occurred prior to a perceptual flip. We also found that changes in horizontal vergence were not responsible for perceptual flips. Thus, eye movements were not essential to flip from one percept to the other. After the moment of a perceptual flip the occurrence probabilities of both saccades and blinks were reduced. The reduced probability of saccades mainly occurred for larger voluntary saccades, rather than for involuntary microsaccades. We suggest that the reduced probability of voluntary saccades reflects a reset of saccade planning. PMID- 15894348 TI - Metabolism of chlorpyrifos in relation to its effect on the availability of some plant nutrients in soil. AB - A laboratory experiment was conducted to study the persistence and metabolism of chlorpyrifos in Gangetic Alluvial soil of West Bengal and also to evaluate their effect on the availability of the major plant nutrients (N, P and K) in soil following the application of chlorpyrifos @ 1 kg (T1), 10 kg (T2) and 100 kg (T3) a.i.ha(-1). The dissipation followed first order kinetics and the calculated half life (T1/2) values ranged from 20 to 37 days. The primary metabolite of chlorpyrifos, 3,5,6-trichloropyridinol (TCP) was detected from 3rd day after application and was at maximum on 30th day which decreased progressively to non detectable level (NDL) on 120th day for all the treatment doses. The secondary metabolite 3,5,6-trichloro-2-methoxy pyridine (TMP) was detected on 30th, 15th and 7th day in T1, T2 and T3 doses respectively which decreased to NDL during 90 120th day. ANOVA study revealed significant decrease in the available N and P content in soil treated with chlorpyrifos in comparison to the control set. The inhibitory effect on available N was attributable to TMP and for P it was due to the presence of TCP and TMP rather than chlorpyrifos itself as revealed by the step wise multiple regression technique. In the later stage of incubation, however the average N and P status was recovered significantly at 120 days which might be due to the disappearance of the metabolites. The variation due to time of observations or treatment doses was minimum in case of available K in soil. PMID- 15894349 TI - Further observations on the primate community at Rudabanya II (late Miocene, early Vallesian age), Hungary. AB - It has been proposed that the pliopithecid Anapithecus hernyaki was hunted by the ape Dryopithecus brancoi based on analyses of the age and sex distribution of 25 individuals from the late Miocene site of Rudabanya, Hungary (ca. 10 Ma). In this study, the minimum number of individuals (MNI) and age distribution of the assemblage are recalculated and considered in relation to their sedimentary context. We suggest that Dryopithecus juveniles may be underrepresented in the assemblage as a consequence of taphonomic loss of deciduous teeth. Furthermore, we find that while Dryopithecus and Anapithecus were contemporaneous, there was little spatial overlap between them, as each taxon is primarily associated with a separate lithofacies. We conclude that when the revised estimates of MNI and age distribution of these primates are reconsidered in conjunction with their context of burial, the case for the Dryopithecus hunting hypothesis cannot be sustained. PMID- 15894350 TI - Total mercury and methylmercury accumulation in periphyton of Boreal Shield lakes: influence of watershed physiographic characteristics. AB - Little is known about Hg accumulation in littoral communities, especially in periphyton biofilm of unperturbed lakes. The objectives of this study were to investigate and establish relationships between total mercury (THg) and methylmercury (MeHg) concentrations in periphyton communities of Boreal lakes and watershed physiographic and lake morphometric characteristics. This study was carried out on 23 Boreal Canadian Shield lakes located between 47-50 degrees N and 73-77 degrees W. Periphyton was sampled on rocks, the dominant periphyton substrate in the littoral zone of these lakes. Periphyton algal biomass (Chla) ranged from 12 to 164 mg m(-2) whereas THg concentrations varied from 42 to 271 ng g(-1) DW and MeHg levels varied from 3 to 55 ng Hg g(-1) DW. Periphyton biomass was positively correlated to latitude, watershed wetland area and negatively correlated to watershed slope and depth of the lake. THg concentrations in periphyton were negatively correlated to watershed wetland area whereas MeHg concentrations were negatively correlated to latitude and positively correlated to watershed slope, dissolved sulfate concentration and the presence of beavers in the lake. This study confirms that periphyton can accumulate large amounts of Hg and the accumulation is strongly influenced by watershed characteristics and periphyton biomass. PMID- 15894351 TI - DDT and human health. AB - 1,1,1-Trichloro-2,2'bis(p-chlorophenyl) ethane (DDT) was the first widely used synthetic pesticide and is extremely persistent in both the environment and the human body. The introduction of DDT revolutionised agricultural production and has been credited with the elimination of malaria from the United States and Europe. However, DDT is also known to have had major environmental consequences and has been associated with dramatic declines in many animal populations. Although DDT use has generally been restricted since the early 1970s, exposure to the pesticide remains widespread. In developed countries, slow elimination from the body means a large proportion of the population still have detectable levels of DDT, or its metabolite DDE, in their serum or adipose tissue. In developing countries, the pesticide continues to be used for vector control and a significant proportion of breast-fed babies has daily intakes above recommended levels. This review considers the epidemiological evidence for possible adverse effects of human exposure to DDT. Much of this research is weakened by methodological flaws. However, recent methods in breast cancer research using nested studies in cohorts with stored biological samples have allowed a more rigorous assessment of a putative role for DDT in disease aetiology. While DDT does not appear to play a causative role in breast cancer development, there is suggestive evidence for a role in the aetiology of other conditions such as pancreatic cancer, neuropsychological dysfunction, and reproductive outcomes. Research into these and other conditions would benefit from the same rigorous approaches used in breast cancer research. Until further high quality evidence is available, it is still too early, even 60 years after the introduction of this once ubiquitous chemical, to pass judgement on the role of DDT in a number of common diseases. PMID- 15894352 TI - Plasminogen interaction with platelets: the importance of carboxyterminal lysines. AB - INTRODUCTION: Thrombin stimulation enhances plasminogen binding to platelets and promotes platelet-dependent plasmin generation. The objective of this study was to determine whether carboxyterminal lysines (C-lysines) are important for these processes, as they are in other cell types. MATERIALS AND METHODS: 125I plasminogen and varying concentrations of unlabeled plasminogen were added to washed platelets that were either resting or stimulated with thrombin, thrombin receptor activating peptide, or ADP. In some experiments the platelets were digested with carboxypeptidase B to remove C-lysines. Platelet-dependent plasmin generation was also studied by adding plasminogen and tissue plasminogen activator to platelet suspensions and monitoring the conversion of a plasmin specific chromogenic substrate. The cells were either resting or stimulated with thrombin, thrombin receptor activating peptide, or ADP. The effect of the thrombin inhibitor lepirudin and the plasmin inhibitor aprotinin on plasminogen binding and the appearance of C-lysines was also investigated. RESULTS: Thrombin, but not thrombin receptor activating peptide or ADP, stimulated high-affinity binding of plasminogen and greatly promoted platelet-dependent plasmin generation. Digestion with carboxypeptidase B eliminated thrombin-induced high affinity binding and reduced thrombin-induced plasmin generation by increasing the Michaelis constant. Lepirudin, but not aprotinin, inhibited thrombin stimulated plasminogen binding to platelets. CONCLUSION: C-terminal lysines are necessary for high-affinity binding of plasminogen to platelets and for platelet supported plasmin generation. The origin of the C-lysines is not clear, but they may result from a direct effect of thrombin, rather than an intermediate enzyme such as plasmin. PMID- 15894353 TI - Upregulation of GP IIb/IIIa receptors during platelet activation: influence on efficacy of receptor blockade. AB - INTRODUCTION: GP IIb/IIIa inhibitor doses high enough to inhibit platelet macroaggregation may not completely prevent formation of microaggregates. Platelets contain an internal pool of GP IIb/IIIa receptors which externalizes with activation and supports microaggregation. This study assesses microaggregation in the presence of the two GP IIb/IIIa inhibitors abciximab and tirofiban. METHODS: Citrated whole blood was preincubated with abciximab (5 microg/ml), tirofiban (50 ng/ml), or saline as control and activated with TRAP (5 microM) or ADP (2 microM) at 37 degrees C under constant stirring. Microaggregate formation and receptor expression were determined with flow cytometry. RESULTS: Within few seconds after TRAP-activation the platelet count dropped from 266,000/microl to 20,000/microl, the number of microaggregates increased from 3700/microl to 10,100/microl and the mean number of GP IIb/IIIa receptors increased from 53,000 to 65,000/platelet. With TRAP+abciximab the platelet count dropped from 259,000 to 113,000/microl, microaggregates increased from 2500 to 9300/microl, GP IIb/IIIa receptors from 56,000 to 77,000/platelet. Platelet microaggregate formation was reversible. With TRAP and tirofiban platelet count dropped to only 190,000/microl, there was no increase in platelet microaggregates, receptors increased to 66,000/platelet. Platelet activation with ADP gave similar results. CONCLUSIONS: During the early phase of activation additional GP IIb/IIIa receptors externalize to the platelet surface. Abciximab does not block these new receptors sufficiently to prevent microaggregate formation. However, the number of unblocked receptors is not high enough to maintain a stable aggregate, microaggregation is reversible. With tirofiban there is no microaggregate formation, possibly because the inhibitor rapidly binds to newly externalized receptors. PMID- 15894354 TI - Squamous cell carcinoma antigen serum levels as prognostic parameter in patients with early stage vulvar cancer. AB - OBJECTIVE: To determine whether SCC-Ag serum levels can be used as a prognostic parameter in surgically treated early stage vulvar cancer. METHODS: SCC-Ag serum levels were measured preoperatively in 61 surgically staged patients with squamous cell vulvar cancer (UICC pT1 and pT2). Results were correlated to clinical data. RESULTS: Mean (standard deviation) SCC-Ag serum levels in patients with vulvar cancer were 1.5 (1.99) ng/mL. SCC-Ag serum levels were significantly higher in patients with pT2 vulvar cancer (2.2 [2.6] ng/mL) compared with patients with pT1 vulvar cancer (1.0 [1.2] ng/mL, P = 0.034). SCC-Ag serum levels were not associated with lymph node involvement (P = 0.1), tumor grade (P = 0.6), and patients' age (P = 0.5). Multivariate Cox regression models considering tumor stage, lymph node involvement, patients' age, and SCC-Ag serum levels as covariates showed that lymph node involvement (P = 0.04 and P = 0.01) and tumor stage (P = 0.006 and P = 0.009), but not SCC-Ag serum levels (P = 0.8 and P = 0.6), and patients' age (P = 0.08 and P = 0.22) are prognostic factors for disease-free and overall survival, respectively. CONCLUSION: SCC-Ag serum levels cannot be used as an additional prognostic parameter in patients with surgically treated early stage vulvar cancer. PMID- 15894355 TI - Identifying patients unlikely to benefit from further chemotherapy: a descriptive study of outcome at each relapse in ovarian cancer. AB - OBJECTIVE: Not all patients with relapsed ovarian cancer (EOC) benefit from further treatment and thus treatment should be selectively applied. METHODS/PATIENTS: A retrospective review of survival and response outcomes in 120 women with relapsed EOC, all treated at original diagnosis with surgery and platin plus paclitaxel, who had all their initial and subsequent relapse therapy carried out at the BCCA. RESULTS: In those patients selected for re-treatment upon relapse, lack of progression rates were 63%, 50%, 45%, 44%, 29%, and 20% respectively for first through sixth relapse. The corresponding median survivals from that relapse were 14, 10, 6, 7, 8, and 5 months. A predictive model based upon the length of the interval between the two preceding relapses (or diagnosis to 2nd relapse) predicted which patients would survive less than 6 months (patient defined "lack of benefit" to chemotherapy criterion): diagnosis to second relapse <12 months; first to third relapse <6 months; second to fourth <6 months; third to fourth <6 months, and fifth to sixth <6 months. CONCLUSION: In selected patients, multiple episodes of re-treatment are of value. A time-based statistic identifies those who will not benefit, defined as survival less than 6 months. PMID- 15894356 TI - Fas gene promoter -670 polymorphism (A/G) is associated with cervical carcinogenesis. AB - OBJECTIVE: To investigate the biological significance of single nucleotide polymorphism (SNP) at Fas gene promoter in cervical carcinogenesis. METHODS: SNP at -670 of Fas gene promoter (A/G) together with human papillomavirus (HPV) types were examined in a total of 279 cervical smear samples and 8 human cervical squamous carcinoma cell lines using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) techniques. RESULTS: 49 patients with high-grade squamous intraepithelial lesion (HSIL) had higher frequency of high risk HPV and GA + GG genotype than 167 with low-grade SIL (LSIL) and 63 controls. G allele frequency was also higher in HSIL than in LSIL and controls. There was an increased OR (6.00; CI, 1.32-27.37; P = 0.021) for GA + GG genotype in HSIL cases compared to controls among 96 patients with high-risk HPV. 7 of 8 cervical carcinoma cell lines also showed GA or GG genotype. CONCLUSION: Fas gene promoter -670 polymorphism (A/G) may be closely associated with cervical carcinogenesis in a Japanese population. PMID- 15894357 TI - Baseline endometrial assessment before tamoxifen for breast cancer in asymptomatic menopausal women. AB - OBJECTIVE: The aim of this study is to estimate the prevalence of endometrial pathology before the start of tamoxifen therapy in menopausal breast cancer patients. METHODS: Ninety-one gynecologically asymptomatic patients, suffering from estrogen receptor-positive breast cancer and scheduled for adjuvant tamoxifen, underwent pretreatment endometrial assessment. In all patients, a transvaginal ultrasonography was carried out; a double-layered endometrial stripe measuring above 4 mm was considered as abnormal. In these patients, outpatient hysteroscopy and endometrial biopsy were performed. Pathologic findings were considered the reference test in estimating the prevalence of endometrial morbidity. RESULTS: In 34 patients (37.3%) a thickened endometrium was an indication for hysteroscopic and pathologic assessment. Endometrial polyps, simple hyperplasias, and complex atypical hyperplasias were found in 10 (10.9%), 4 (4.3%), and 3 (3.2%) patients, respectively, leading to an overall prevalence of baseline endometrial morbidity of 18.6%. Established individual risk factors for development of endometrial pathology, such as body mass index, age at menarche and menopause, and parity, did not significantly differ in patients with and without endometrial abnormalities. Only patients' age (63.8 +/- 8.6 and 52.2 +/- 11.8; P = 0.03) and endometrial thickness (10.5 +/- 3.5 and 3.9 +/- 3.0; P > 0.001) were significant predictive factors of endometrial pathology. CONCLUSIONS: Menopausal women with estrogen receptor-positive breast cancer appear to have high risk of baseline subclinical endometrial abnormalities; therefore, an endometrial assessment, before the start of tamoxifen therapy, is always recommended in such patients. PMID- 15894358 TI - Cyclins D1, D3, E, and A in vulvar carcinoma patients. AB - OBJECTIVE: The treatment of vulvar squamous cell carcinoma patients is often mutilating. Effort is being made to individualize treatment in order to reduce negative side effects for patients with good prognosis. Molecular markers have been able to predict patient outcome in several tumors. The aim of this study was to characterize the expression of cyclins D1, D3, E, and A in a comparatively large series of patients with vulvar squamous cell carcinoma and look for prognostic impact. METHODS: A total of 224 vulvar squamous cell carcinomas were immunohistochemically investigated for expression of cyclins D1, D3, E, and A using the biotin-streptavidin-peroxidase method and the OptiMax Plus automated cell staining system. RESULTS: High protein levels of cyclin D1 (any positive nuclei) were found in 58 (26%) cases, cyclin D3 (> or =50% positive nuclei) in 61 (27%) cases, cyclin E (> or =50% positive nuclei) in 41 (18%) cases, and cyclin A (> or =5% positive nuclei) in 156 (70%) cases. No prognostic impact was found for the cyclins D1, D3, E, or A. CONCLUSIONS: The high number of cases showing increased levels of cyclin A suggests that this protein may be important in the pathogenesis of vulvar squamous cell carcinoma. No prognostic impact was found for the cyclins D1, D3, E, or A. PMID- 15894359 TI - Immunosuppressive activity of proteases in cervical carcinoma. AB - OBJECTIVE: The host immune response is essential for restraining both HPV infections and HPV-related cervical cancer. We previously reported a direct correlation between proteolytic activity and malignant progression from precursor lesions to invasive cervical carcinoma. The present study was undertaken to investigate whether proteinases from cervical carcinoma extracts and representative purified proteinases involved in tumor progression could regulate lymphocyte proliferation to phytohemagglutinin (PHA) mitogen. METHODS: Extracts were prepared from tissue samples obtained from patients with invasive cervical squamous carcinoma, squamous intra-epithelial lesions or women with normal cervix. Lymphocytes obtained from a single healthy donor were pre-incubated with one of these extracts in the presence or absence of proteinase inhibitors, and stimulated with PHA during 72 h. The proliferative response was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) method (re-validated with thymidine uptake). RESULTS: Lymphocyte proliferation was significantly decreased by cervical carcinoma extracts, while only slightly decreased by squamous intra-epithelial lesions or normal extracts. Inhibitor assays indicated that proteinases from cervical carcinoma were responsible for 53.30% of total suppressive activity. We found that purified enzymes such as trypsin, cathepsin B, uPA and type IV collagenase suppressed the proliferative response in a dose-dependent fashion. CONCLUSIONS: Our data suggest that in addition to the classic role in tumor invasion, proteases could represent an immune evasion mechanism in cervical carcinoma. PMID- 15894360 TI - A multicenter phase II study of gemcitabine, paclitaxel, and cisplatin in chemonaive advanced ovarian cancer. AB - OBJECTIVES: The objectives of this multicenter phase II study were to evaluate the effects of gemcitabine-paclitaxel-cisplatin combination chemotherapy on response rate, survival, and toxicity in patients with advanced epithelial ovarian cancer (AEOC). METHODS: Chemonaive AEOC patients with bidimensionally measurable disease or an elevated serum cancer antigen 125 level received cisplatin (70 mg/m(2)) on day 1 and paclitaxel (80 mg/m(2)) and gemcitabine (1000 mg/m(2)) on days 1 and 8, every 3 weeks. RESULTS: Between October 2000 and September 2001, 46 patients were enrolled. Sixteen patients underwent debulking surgery prior to chemotherapy. In 45 evaluable patients, overall response rate was 64.4% (7 CR and 22 PR). Median time-to-progression was 13.4 months (95% CI, 9.6-17.4 months); median progression-free survival was 12.3 months (95% CI, 8.8 15.6 months); median overall survival was 26.0 months (95% CI, 18 months-not reached); and 1-year survival was 74% (95% CI, 60-88%). The relative dose intensities of gemcitabine, paclitaxel, and cisplatin were 81.4%, 80.2%, and 89.8%, respectively. Grade 3/4 neutropenia was the predominant hematologic toxicity observed (73.9% of patients) followed by grade 3/4 leukopenia (56.5%), anemia (45.7%), thrombocytopenia (23.9%), and febrile neutropenia/neutropenic sepsis (26.1%). The predominant grade 3 nonhematologic toxicities were alopecia (43.5%) and diarrhea (19.6%). Grade 4 nonhematologic toxicities were nausea/vomiting, constipation, and uremia (2.2% each). Two treatment-related deaths occurred (neutropenic sepsis and uremia). CONCLUSION: Gemcitabine paclitaxel-cisplatin combination chemotherapy is active with manageable toxicity in chemonaive patients with advanced ovarian cancer and should be explored in larger phase III trials. PMID- 15894361 TI - Phase one dose finding study of capecitabine (Xeloda), radiotherapy and cisplatin in the treatment of locally advanced squamous cervical cancer. AB - OBJECTIVE: A dose escalation study was performed for patients with locally advanced squamous carcinoma of cervix to determine the maximum tolerated dose (MTD) of daily capecitabine when combined with weekly cisplatin and radiotherapy. MTD would be reached if at least 2 out of 6 at any particular dose level developed Grade 3 or 4 diarrhoea, mucositis, skin or bladder toxicity or Grade 4 neutropenia associated with fever lasting more than 7 days or Grade 3 or 4 thrombocytopenia. The secondary endpoints were to define the toxicity profile, compliance with treatment, late radiation effects and to determine the tumour responses. METHODS: Thirteen patients were enrolled (7 Stage II(b), 6 Stage III(b)). Cisplatin was given weekly at 40 mg/m2 (maximum 70 mg) concurrently with radiotherapy. Two doses of capecitabine were studied 300 mg/m2 bid (cohort 1) and 450 mg bid (cohort 2). RESULTS: One patient in cohort 1 developed Grade 3 diarrhoea and 2 patients had Grade 3 leucopenia. Two patients in cohort 2 developed Grade 4 leucopenia and neutropenic fever; one of these patients also had Grade 3 diarrhoea and thrombocytopenia. Only 1 patient had Grade 1 palmar plantar erysthrodysesthesia. Three patients have developed late (RTOG/ECOG) Grade 3 toxicity bladder or vaginal mucosa at 6, 9 and 15 months. The MDT of continuous capecitabine, when given with pelvic radiotherapy and weekly cisplatin, is 450 mg m2 bid. CONCLUSIONS: The recommended dose of capecitabine in this combination is 300 mg/m2 bid. The regimen was well tolerated and the compliance was high. Progression-free survival at 12 months was 69.2%, and at 24 months, it was 49.2% (SE 15.4%) with an overall survival of 57.7% (SE 15.0%) at 24 months. Further evaluation of this regimen in a phase II/III study is warranted. PMID- 15894362 TI - Determination of HER2/neu status in uterine serous papillary carcinoma: Comparative analysis of immunohistochemistry and fluorescence in situ hybridization. AB - OBJECTIVE: To evaluate and compare HER2/neu protein overexpression and gene amplification in uterine serous papillary endometrial cancer (USPC). STUDY DESIGN: Immunohistochemical (IHC) and fluorescent in situ hybridization (FISH) assays were used to analyze and compare HER2/neu protein expression and gene amplification, respectively, in paraffin blocks from 26 women harboring stage IA to IV USPC treated at the University of Arkansas for Medical Sciences from 1997 to 2004. Chromosome 17 polysomy status by FISH was also assessed in all specimens. RESULTS: Moderate-to-strong expression of HER2/neu protein was noted in 16 (62%) of 26 USPC samples evaluated, with 7 (27%) samples showing moderate staining (2+) and 9 (35%) showing strong staining (3+) for HER2/neu. Amplification of the ERBB2 gene by FISH was observed in 11 of the 26 (42%) cases. Protein overexpression and gene amplification were found to correlate in 100% (9 of 9) of the 3+ positive tumors and 2 out of 7 (29%) of the 2+ positive tumors. Heterogeneity was noted in 3 cases in the amplification of the HER2/neu gene within the same tumor samples with pockets of amplified tumor cells amidst nonamplified tumor cells. None of the 10 USPC cases scored by IHC as 0 or 1+ was found positive for ERBB2 amplification by FISH. CONCLUSIONS: Amplification of the HER2/neu oncogene represents a common finding in USPC. FISH analysis should be used for confirmation of gene amplification in USPC showing 2+ expression of HER2/neu. Prior screening and selection of appropriate immunohistochemistry positive areas may be beneficial in the selection of some USPC patients undergoing FISH analysis. PMID- 15894363 TI - Predictors of vaginal relapse in stage I endometrial cancer. AB - OBJECTIVE: To identify factors predictive of vaginal relapse in stage I endometrial cancer, thereby potentially facilitating the selection of patients who may benefit from vaginal brachytherapy. METHODS: The study population included 632 patients with stage I endometrial cancer managed with hysterectomy at our institution between 1984 and 1996. Median follow-up was 73 months; 122 patients (19%) received adjuvant radiotherapy. RESULTS: Overall, 2.9% of the stage I cohort developed vaginal relapse at 5 years. Vaginal relapse was observed in 1.7% of patients who received radiotherapy and in 3.0% of those whose treatment did not include radiotherapy (P = 0.36). Cox regression analysis (including radiotherapy) identified only grade 3 differentiation (hazard ratio = 3.83, P = 0.007) as an independent predictor of vaginal relapse. Patients with a low-grade tumor had a 5-year vaginal relapse rate of 2%, compared with 7% for those with a grade 3 tumor. When only patients who did not receive adjuvant radiotherapy were considered, both grade 3 tumor and lymphovascular invasion were significant predictors of vaginal relapse (P < 0.05). When neither variable was present, 2% of patients experienced vaginal relapse at 5 years, compared with 11% when either 1 was present (P < 0.001). Depth of myometrial invasion was not a significant predictor of vaginal recurrence. CONCLUSION: Histologic grade 3 tumor and lymphovascular invasion were the cogent predictors of vaginal relapse in our population. The cost and morbidity of vaginal brachytherapy should be balanced against the potential risk of vaginal relapse in this group of patients. PMID- 15894364 TI - Human papillomavirus type and tobacco use as predictors of survival in early stage cervical carcinoma. AB - OBJECTIVE: Molecular and environmental co-factors are known risk factors for cervical cancer. The aim of this study was to define the prognostic significance of HPV 18 and its phylogenetically related viruses and smoking on survival in patients with early stage cervical cancer. METHODS: HPV typing was performed on stage IB-IIB cervical tumors. Subjects positive for HPV 18 or 45 were compared to the remainder of the cohort and to women with tumors containing HPV 16, 31, or 52. Tobacco use was ascertained by patient questionnaire. RESULTS: Tumors of 255 women were evaluated. The presence of HPV 18 or 45 was associated with decreased survival. In a multivariable Cox proportional hazards analysis comparing patients with HPV 18 or 45 containing tumors to the rest of the cohort, the hazard ratio for death from cervical cancer was 2.08 (95% CI, 1.07-4.04). The hazard ratio for death from cervical cancer was 2.41 (95% CI, 1.17-4.96) when the HPV 18 and 45 group was compared to women with HPV 16 or its related viruses, 31 and 52. Smoking was associated with a decreased survival for women with HPV 18 or 45, even after adjusting for other known prognostic factors (P = 0.031). CONCLUSIONS: In addition to pathologic indicators, molecular and environmental co-factors are important determinates of outcome in early stage cervical cancer. The presence of HPV 18 or 45 is associated with a decreased survival. The adverse effect of HPV 18 and 45 on survival is compounded by tobacco use. PMID- 15894365 TI - Demethylation of the MCJ gene in stage III/IV epithelial ovarian cancer and response to chemotherapy. AB - OBJECTIVE: Methylation of a CpG island within the Methylation controlled DNAJ (MCJ) gene results in loss of expression in normal and neoplastic cells. Normal ovarian surface epithelial cells are methylated at the MCJ CpG island and do not express the MCJ gene. Furthermore, re-expression of the MCJ gene, in ovarian cancer cell lines, has been correlated with increased sensitivity to several important chemotherapeutic drugs. The objective of this study was to determine the extent of MCJ promoter methylation in epithelial ovarian cancer patients and address the possible role of MCJ methylation levels in response to chemotherapy in ovarian cancer patients. METHODS: The methylation status of 35 CpG sites within the MCJ CpG island was determined by sequencing of sodium bisulfite modified tumor DNA in 41 patients with stage III/IV epithelial ovarian tumors. Levels of methylation of the MCJ CpG island were then compared with response to therapy and overall survival in the patients. RESULTS: The analysis identified frequent loss of MCJ methylation in ovarian tumors, with only a subset retaining high methylation levels. While 93% (38/41) of tumors examined showed some level of MCJ methylation, only 17% (7/41) retained very high levels (>90% methylation). The presence of such high levels of CpG island methylation correlated significantly with poor response of patients' tumors to therapy (P = 0.027) and poor overall survival (P = 0.023, hazard ratio = 2.9). CONCLUSIONS: These results suggest that MCJ methylation may be useful as a marker of response to chemotherapy in ovarian cancer and are consistent with previous in vitro data linking loss of MCJ expression with drug resistance. PMID- 15894366 TI - Comparison between conventional surgery and radiotherapy for FIGO stage I-II cervical carcinoma: a retrospective Japanese study. AB - PURPOSE: To compare treatment outcome results of conventional surgery vs. radiotherapy (RT) for carcinoma of the uterine cervix. MATERIALS AND METHODS: A retrospective analysis was conducted of 152 patients with uterine cervical cancer radically treated with surgery or high dose-rate intracavitary brachytherapy (HDR ICBT) with or without external RT from June 1991 to May 2004. The median follow up time was 43.5 months (range, 1.0-130.0 months). The median age was 53 years (range, 25-81 years). There were 13 patients (9%) in stage IA, 52 (34%) in stage IB, 24 (16%) in stage IIA, and 63 (41%) in stage IIB. The conventional surgery group included 115 patients (76%) who underwent hysterectomy with pelvic lymph node dissection. Of these, 72 (63%) received postoperative radiotherapy. Thirty seven patients (24%) were assigned to the RT group. Of these, 14 (38%) received chemoradiotherapy. Three patients with stage I received ICBT-alone without external beam irradiation. RESULTS: The 5-year cause-specific survival (CSS) rates for surgery and RT were 79.9% and 82.3%, respectively; the difference between these two treatments was not statistically significant (P = 0.8524). The differences in the survival rates between the two treatments for each of the stage I or stage II patients were also not statistically significant (P = 0.8407 for stage I and P = 0.6418 for stage II). CONCLUSIONS: This retrospective study suggests that RT results in compatible survival with conventional surgery for patients with stage I-II cervical carcinoma. PMID- 15894367 TI - Psychosocial factors and the course of cervical intra-epithelial neoplasia: a prospective study. AB - OBJECTIVE: To investigate the influence of psychosocial factors on the course of cervical intra-epithelial neoplasia (CIN). METHODS: A group of 93 patients with CIN 1 or 2 was followed for 2.25 years by half-yearly colposcopy and cytology. Negatively-rated life events, social support, and coping style were studied in relation to distress during follow-up and in relation to time till progression and regression of CIN. Human papillomavirus (HPV) infection was controlled for as well as sick role bias caused by suspicion of having cervical cancer and distress due to the abnormal cervical smear. RESULTS: During follow-up, progression was found in 20 patients (22%), stable disease in 22 patients (24%), and regression in 51 patients (55%). Negatively-rated life events and lack of social support predicted distress longitudinally. No association was found between progression or regression of CIN and negatively-rated life events, lack of social support, coping style, and distress. CONCLUSION: We found no evidence that psychosocial factors influence the course of CIN. PMID- 15894368 TI - Well-differentiated papillary mesothelioma of the peritoneum: a pathological analysis and review of the literature. AB - BACKGROUND: Well-differentiated papillary mesothelioma (WDPM) of the peritoneum is a rare subtype of peritoneal epithelioid mesothelioma which typically has low malignant potential. It most commonly occurs in young women lacking a history of asbestos exposure. Only 38 female patients with peritoneal WPDM have been reported in the literature, and no uniform treatment recommendation has been established. CASE REPORT: A 74-year-old asymptomatic woman without significant past medical history underwent workup and subsequent surgery for an adnexal mass with a normal serum CA-125 level. Exploratory laparotomy identified an ovarian serous cystadenoma and an incidental multifocal peritoneal neoplasm with extensive calcifications. Histology and cytology confirmed WDPM with extensive, intimately associated mesothelial cystic inclusions and zonal calcifications with osseous metaplasia. Our patient did not receive adjuvant therapy and was without clinical or radiologic evidence of disease 12 months after diagnosis. CONCLUSION: WDPM of the peritoneum in women is frequently asymptomatic and associated with an indolent course. Patient outcomes are usually favorable after tumor-debulking surgery without adjuvant therapy. PMID- 15894369 TI - Irinotecan hydrochloride (CPT-11) and mitomycin C as the first line chemotherapy for ovarian clear cell adenocarcinoma. AB - OBJECTIVE: The purpose of this study was to report the results of adjuvant CPT-11 and MMC combination chemotherapy (CPT-M) for ovarian clear cell adenocarcinoma (OCCA). METHODS: Between 1996 and 2002, 20 patients with OCCA underwent primary debulking surgery and received 6 treatments of CPT-11 (140 mg/m2) in combination with MMC (7 mg/m2), 2 weeks apart with a space of 3-4 weeks between the 3rd and 4th treatment in adjuvant setting. Overall survival was compared with our historical control treated between 1983 and 1995, in which 14 patients with OCCA were treated with an initial optimal standard surgery and postoperative adjuvant cyclophosphamide, doxorubicin, and cisplatin (CAP) combination chemotherapy. RESULTS: Median age was 51 years old (range, 29-74). Twelve patients were in stage Ic, 1 in stage IIa, 5 in stage IIc, 1 in stage IIIc, and 1 in stage IV. Optimal cytoreduction with standard surgery was obtained in all 20 patients. The major toxicity with this regimen was neutropenia, which was reversible. The incidences of grade 3 and 4 neutropenia were 25% and 15%, respectively. The non hematological toxicities were generally mild and well tolerated. One patient with stage Ic refused chemotherapy after the first cycle of CPT-M, and died of her disease 8 months after initial surgery. Five-year survival rate was 95.0% for CPT M group, and 63.5% for CAP group (P = 0.042). Survival was significantly better for patients treated with CPT-M. CONCLUSION: This preliminary study shows that the combination of CPT-M appears to be safe and useful in patients with OCCA. Prospective randomized trials should be conducted to assess this regimen appropriate for women with OCCA. PMID- 15894370 TI - Chitosan: a versatile biopolymer for orthopaedic tissue-engineering. AB - Current tissue engineering strategies are focused on the restoration of pathologically altered tissue architecture by transplantation of cells in combination with supportive scaffolds and biomolecules. In recent years, considerable attention has been given to chitosan (CS)-based materials and their applications in the field of orthopedic tissue engineering. Interesting characteristics that render chitosan suitable for this purpose are a minimal foreign body reaction, an intrinsic antibacterial nature, and the ability to be molded in various geometries and forms such as porous structures, suitable for cell ingrowth and osteoconduction. Due to its favorable gelling properties chitosan can deliver morphogenic factors and pharmaceutical agents in a controlled fashion. Its cationic nature allows it to complex DNA molecules making it an ideal candidate for gene delivery strategies. The ability to manipulate and reconstitute tissue structure and function using this material has tremendous clinical implications and is likely to play a key role in cell and gene therapies in coming years. In this paper we will review the current applications and future directions of CS in articular cartilage, intervertebral disk and bone tissue engineering. PMID- 15894371 TI - Electrospun protein fibers as matrices for tissue engineering. AB - Electrospinning has recently emerged as a leading technique for generating biomimetic scaffolds made of synthetic and natural polymers for tissue engineering applications. In this study, we compared collagen, gelatin (denatured collagen), solubilized alpha-elastin, and, as a first, recombinant human tropoelastin as biopolymeric materials for fabricating tissue engineered scaffolds by electrospinning. In extending previous studies, we optimized the shape and size (diameter or width) of the ensuing electrospun fibers by varying important parameters of the electrospinning process, such as solute concentration and delivery rate of the polymers. Our results indicate that the average diameter of gelatin and collagen fibers could be scaled down to 200-500 nm without any beads, while the alpha-elastin and tropoelastin fibers were several microns in width. Importantly, and contrary to any hitherto reported structures of electrospun polymers, fibers composed of alpha-elastin, especially tropoelastin, exhibited "quasi-elastic" wave-like patterns at increased solution delivery rates. The periodicity of these wave-like tropoelastin fibers was partly affected by the delivery rate. Atomic force microscopy was utilized to profile the topography of individual electrospun fibers and microtensile testing was performed to measure their mechanical properties. Cell culture studies confirmed that the electrospun engineered protein scaffolds support attachment and growth of human embryonic palatal mesenchymal (HEPM) cells. PMID- 15894372 TI - Poly(d,l-lactide-co-glycolide)/montmorillonite nanoparticles for oral delivery of anticancer drugs. AB - This research developed a novel bioadhesive drug delivery system, poly(d,l lactide-co-glycolide)/montmorillonite (PLGA/MMT) nanoparticles, for oral delivery of paclitaxel. Paclitaxel-loaded PLGA/MMT nanoparticles were prepared by the emulsion/solvent evaporation method. MMT was incorporated in the formulation as a matrix material component, which also plays the role of a co-emulsifier in the nanoparticle preparation process. Paclitaxel-loaded PLGA/MMT nanoparticles were found to be of spherical shape with a mean size of around 310 nm and polydispersity of less than 0.150. Adding MMT component to the matrix material appears to have little influence on the particles size and the drug encapsulation efficiency. The drug release pattern was found biphasic with an initial burst followed by a slow, sustained release, which was not remarkably affected by the MMT component. Cellular uptake of the fluorescent coumarin 6-loaded PLGA/MMT nanoparticles showed that MMT enhanced the cellular uptake efficiency of the pure PLGA nanoparticles by 57-177% for Caco-2 cells and 11-55% for HT-29 cells, which was dependent on the amount of MMT and the particle concentration in incubation. Such a novel formulation is expected to possess extended residence time in the gastrointestinal (GI) tract, which promotes oral delivery of paclitaxel. PMID- 15894373 TI - Re: It is a mistake to conclude that sexual abuse and sexualized behavior are not related: a reply to Drach, Wientzen, and Ricci (2001). PMID- 15894374 TI - Inhibitory control of the acute mu-withdrawal response by indirectly activated adenosine A1 and kappa-opioid systems in the Guinea-pig ileum; reversal by cholecystokinin. AB - In the isolated guinea-pig ileum (GPI), the acute mu-opioid withdrawal response is inhibited by the kappa-opioid system, indirectly activated by the opioid agonist; yet, other inhibitory mechanisms are probably operating. On the other hand, cholecystokinin (CCK-8) strongly enhances the withdrawal response. In this study, we have shown that the adenosine A1 antagonist 8-cyclopenthyl-1,3 dimethylxantine (CPT) increased the withdrawal response in dermorphin/naloxone (NLX) tests but lacked any effect if the withdrawal tests were carried out in presence of CCK-8. In tissue preparations coming from a same animal both CPT and the kappa-opioid antagonist, nor-binaltorphimine (BNI), increased the intensity of the withdrawal responses; the effects of the two antagonists were additive. The intensity of withdrawal contractile responses in presence of CCK-8 was similar to those obtained in presence of the two antagonists. Tissue preparations tested with dermorphin/CCK-8/NLX and then washed out yielded contractile responses when subsequently challenged with CPT, BNI or BNI+CPT, with a percentage markedly higher than the percentage of the response to NLX challenge. BNI+CPT also increased the intensity of the response to NLX challenge. These data suggest that acute exposure of GPI to dermorphin induces the activation of both the adenosine A1 and kappa-opioid systems, which in turns inhibit the mu withdrawal response. CCK-8 antagonises the inhibitory effect of the indirectly activated systems. PMID- 15894375 TI - A cellular system to study the role of nitric oxide in cell death, survival, and migration. AB - The gaseous messenger nitric oxide (NO) plays a bewildering number of roles in fundamental processes, such as cell locomotion, differentiation, proliferation and death. Its different and often contrasting roles may depend on its concentration and intracellular site of generation. We describe here a simple system with which to investigate the roles of NO generated at physiological levels in HeLa cells by eNOS transfected under an inducible promoter. This system has allowed us to uncover unexpected signalling circuits between NO and ceramide, involved in the response of cells to apoptotic stimuli. At present, we are using these cells as a tool to investigate the role of NO in migration. PMID- 15894376 TI - Antioxidant treatment inhibited glutamate-evoked NF-kappaB activation in primary astroglial cell cultures. AB - In glial cells, glutamate exposure causes alterations in cell redox status, mainly mediated by glutathione depletion and reactive oxygen species generation. These effects finally lead to astrocyte dysfunction which contributes to the pathogenesis of several neurological disorders. This study was aimed to investigate the involvement of the NF-kappaB pathway in oxidative stress induced by glutamate exposure in primary cultures of astrocytes. Further, we evaluated the power of the antioxidants genistein (0.1-10 microM) and IRFI 016 (20-80 microM), a synthetic tocopherol analogue, compared with glutathione ethyl ester (10-50 microM) and cysteamine-HCl (100-500 microM), to antagonize the effects elicited by glutamate (500 microM). Alterations of cell redox status were reduced, in a dose-dependent way, by antioxidants; in particular, 80 microM IRFI 016 and 10 microM genistein almost completely restored glutathione basal levels and significantly diminished ROS production, as well as 100 microM glutathione ethyl ester. These antioxidant effects were stronger than those caused by 500 microM cysteamine-HCl. Further, glutamate promoted the up-regulation of p50 and p65 NF-kappaB subunits and their nuclear translocation, as revealed by Western blot analysis and electrophoretic mobility shift assay of both subunits. The activation of p50 and p65 NF-kappaB subunits induced by glutamate exposure was significantly reduced by IRFI 016, acting in a dose-dependent manner. Altogether, these data confirm that the NF-kappaB pathway is involved in cell response to oxidative stress induced by glutamate injury in primary astrocyte cultures, and suggest that the use of antioxidants, such as IRFI 016, may be a helpful pharmacological strategy for neuroprotection. PMID- 15894377 TI - Cu(II) complexes with a sulfonamide derived from benzoguanamine. Oxidative cleavage of DNA in the presence of H2O2 and ascorbate. AB - Reaction between benzoguanamine (2,4-diamino-6-phenyl-1,3,5-triazine) and 2 mesitylenesulfonyl chloride leads to formation of a sulfonamide able to form two mononuclear Cu(II) complexes with a CuL(2) stoichiometry. The local environment of the metal cation is a distorted octahedron, with two ligands and two solvent molecules; both complexes crystallize in the monoclinic structure, space group P2(1), with Z=2. In the presence of ascorbate/H(2)O(2,) the two complexes significantly cleavage double-strand pUC18 DNA plasmid. Both complexes exhibit more nuclease efficiency that the copper phenantroline complex. From scavenging reactive oxygen studies we conclude that the hydroxyl radical and a singlet oxygen-like entity, such a peroxide copper complex, are the radical species involved in the DNA damage. PMID- 15894378 TI - The clinical potential of chemokine receptor antagonists. AB - Chemokines belong to a family of chemotactic cytokines that direct the migration of immune cells towards sites of inflammation. They mediate their biological effects by binding to cell surface receptors, which belong to the G protein coupled receptor superfamily. Since chemokines and their receptors have been implicated in the pathophysiology of a number of autoinflammatory diseases, chemokine receptor antagonists could prove to be useful therapeutics to target these diseases. Here, we review the role of chemokines in autoimmunity, concentrating mainly on the chemokine receptors CCR1 and CCR5, and discuss the potential utility of antagonists that target these 2 receptors as they progress through the clinic. PMID- 15894379 TI - Applying resampling methods to neurophysiological data. AB - Standard statistical techniques do not always provide answers to complex physiological questions because often there are no parametric or non-parametric distributions on which significance can be estimated. Resampling methods provide a battery of tests that can be used in such circumstances. In the past few years these methods have been explored theoretically and are now employed frequently. In this paper we describe a unified framework for the use of such methods in the context of neurophysiological data analysis. We construct specific tests for placing confidence limits on estimates of mutual information and on parameters of circular data, and we present procedures for testing hypotheses on circular and on partitioned data. These tests are explained in detail and illustrated with real data from experiments with behaving monkeys. PMID- 15894380 TI - Panic disorder and bipolar disorder: anxiety sensitivity as a potential mediator of panic during manic states. AB - BACKGROUND: Panic disorder (PD) occurs at high rates in bipolar disorder and more commonly than in unipolar depression. Reports of PD onset during hypomania and depressive mania (i.e., mixed states) raise questions about whether the affective disturbances of bipolar disorder play a specific role in the exacerbation or onset of PD. Anxiety sensitivity (AS), a risk factor for PD appears greater in bipolar disorder compared to unipolar depression, although the association of specific mood states with AS remains unknown. METHODS: We examined the association of current mood state (i.e., mixed state, mania or hypomania, bipolar depression, unipolar depression, and euthymia) with Anxiety Sensitivity Index (ASI) scores in 202 individuals with bipolar disorder (n=110) or major depressive disorder (n=92). RESULTS: Current mood state was significantly associated with ASI score (Chi-square=21.2, df=4, p=0.0003). In multiple regression analyses, including covariates for comorbid anxiety disorders, current mania or hypomania was a significant predictor of ASI scores (p<0.04). Current mixed state tended toward a similar association (p<0.10). LIMITATIONS: Conclusions are limited by the study's cross-sectional nature and relatively small sample size. CONCLUSIONS: These findings of elevated AS during manic states, independent of comorbid anxiety disorders, provide preliminary support for the hypothesis that manic states contribute to risk for the development or exacerbation of PD, and that AS may contribute to the high prevalence and severity of PD comorbid with bipolar disorder. PMID- 15894381 TI - Comorbid psychiatric disorders in depressed outpatients: demographic and clinical features. AB - BACKGROUND: This study evaluated the clinical and sociodemographic features associated with various degrees of concurrent comorbidity in adult outpatients with nonpsychotic major depressive disorder (MDD). METHODS: Outpatients enrolled in the STAR*D trial completed the Psychiatric Diagnostic Screening Questionnaire (PDSQ). An a priori 90% specificity threshold was set for PDSQ responses to ascertain the presence of 11 different concurrent DSM-IV Axis I disorders. RESULTS: Of 1376 outpatients, 38.2% had no concurrent comorbidities, while 25.6% suffered one, 16.1% suffered two, and 20.2% suffered three or more comorbid conditions. Altogether, 29.3% met threshold for social anxiety disorder, 20.8% for generalized anxiety disorder, 18.8% for posttraumatic stress disorder, 12.4% for bulimia, 11.9% for alcohol abuse/dependence, 13.4% for obsessive-compulsive disorder, 11.1% for panic disorder, 9.4% for agoraphobia, 7.3% for drug abuse/dependence, 3.7% for hypochondriasis, and 2.2% for somatoform disorder. Those with more concurrent Axis I conditions had earlier ages at first onset of MDD, longer histories of MDD, greater depressive symptom severity, more general medical comorbidity (even though they were younger than those with fewer comorbid conditions), poorer physical and mental function, health perceptions, and life satisfaction; and were more likely to be seen in primary care settings. LIMITATIONS: Participants had to meet entry criteria for STAR*D. Ascertainment of comorbid conditions was not based on a structured interview. CONCLUSIONS: Concurrent Axis I conditions (most often anxiety disorders) are very common with MDD. Greater numbers of concurrent comorbid conditions were associated with increased severity, morbidity, and chronicity of their MDD. PMID- 15894382 TI - Is "relationship fear" unique to Japan? Symptom factors and patient clusters of social anxiety disorder among the Japanese clinical population. AB - BACKGROUND: There have been only a limited number of cross-cultural studies of social anxiety disorder (SAD), especially as diagnosed with modern operationalized diagnostic criteria and as measured with recently developed assessment instruments. We examined the symptomatological structure and clinical subtypes of patients with DSM-IV SAD among the Japanese clinical population. METHODS: We performed confirmatory and exploratory factor analyses of the joint Social Interaction Anxiety Scale and Social Phobia Scale from 149 psychiatric patients diagnosed with SAD. Based on the derived symptom factors, we further ran cluster analysis to identify patient subgroups. RESULTS: Factor analyses revealed three factors which were named "scrutiny fears", "conversation fears" and "relationship fears". The first two appeared common to Western clinical populations but the third appeared unique to the Japanese. Cluster analysis based on these three factor scores yielded three subgroups, which were externally validated and which overall corresponded with mild, moderate and pervasive subtypes of social phobia. LIMITATIONS: Both factor analysis and cluster analysis employed in the present study are exploratory in nature. Further empirical examination in different settings and cultures is necessary to provide definitive answers. CONCLUSIONS: It is suggested that we may need three symptom subscales and three subtypes in order to better account for cultural variations in the presentation of SAD. PMID- 15894383 TI - "Altered-self" or "near-self" in the positive selection of lymphocyte repertoires? AB - Positive selection of lymphocyte repertoires is now recognized as applying to both B and T cells. However, much of the early literature on positive selection focussed on cell-mediated immunity (T cells), which biased consideration of its general biological role. The term "altered-self," which initially captured the idea of self (i.e. MHC) altered by the addition of what was later found to be a peptide fragment, has not proven robust and may now be clouding our understanding. It is recommended that the term "near-self" be reintroduced since it captures the essence of the probable underlying adaptive process-sub-threshold self-reactivity to countermand rapid pathogen mutation. PMID- 15894384 TI - Repeated attacks of venous air embolism during endoscopic sinus tumor surgery: a case report. AB - Repeated attacks of venous air embolism during an operation is a very rare situation. We report a case of multiple venous air emboli during an endoscopic surgery of a sphenoidal sinus tumor. PMID- 15894385 TI - Normalization of maxillary sinus mucosa after functional endoscopic sinus surgery in pediatric chronic sinusitis. AB - OBJECTIVES: To observe the recovery of maxillary sinus mucosa after endoscopic sinus surgery in children with chronic maxillary sinusitis. Functional endoscopic sinus surgery (FESS) was used to perform a middle meatal antrostomy, leaving the antral mucosa intact for later observation. METHODS: Between January 1998 and December 2003, 43 children with chronic sinusitis were enrolled in the study. Pre operative patient profiles, including a history of symptoms, signs and allergies, were collected. Phidiatap test was used to check allergy. Saccharine transit time tests were performed for each side of the nasal cavity. Under endoscopic observation, the antral mucosa was macroscopically divided into edematous and polypoid types. RESULTS: Sixty-four (76.2%) of the 84 antrums had edematous type mucosa in which 73.4% (48/64) of cases were found to have returned to normal within 8 weeks. The polypoid antral mucosa exhibited a slower recovery with 80% (16/20) returning to normal within 4 months. The preoperative saccharine transit time significantly correlated with recovery of the antral mucosa (p < 0.05), but allergy did not (p > 0.05). CONCLUSION: The antral mucosa in children with chronic maxillary sinusitis was predominantly of the edematous type. Most recovered within 2 months of having FESS. The prolonged saccharine transit time and polypoid type antral mucosa were associated with delayed mucosal recovery, warranting follow-up of more than 4 months. PMID- 15894386 TI - Clivus osteomyelitis secondary to Enterococcus faecium infection in a 6-year-old girl. AB - A 6-year-old girl was diagnosed to suffer from clivus osteomyelitis secondary to Enterococcus faecium infection. On the basis of the magnetic resonance image, the abscess was drained via the posterior wall of the pharyngeal tract immediately. Subsequent antibiotic therapy allowed rapid improvement and long-term healing of the osteomyelitic process without any side effect. Osteomyelitis or abscess of the clivus is very rare in adult patients and extremely rare in children. Some etiopathogenetic hypotheses are discussed in this case. PMID- 15894387 TI - Visualization of discrete L1 oligomers in human papillomavirus 16 virus-like particles by gel electrophoresis with Coomassie staining. AB - The recombinant major capsid protein (L1) of human papillomavirus (HPV) can self assemble into virus-like particles (VLPs) with 360 L1 molecules per VLP. These tightly associated L1 oligomers in the assembled VLPs were disrupted in a pH-, denaturant-, time-, and temperature-dependent fashion. With non-reducing Laemmli type SDS-PAGE, primarily the monomeric L1 protein ( approximately 55 kDa) is observed when analyzing VLP preparations. When the pH was lowered to pH 7.0 in NuPAGE system and the gel temperature during electrophoresis was maintained at a lower temperature ( approximately 7 degrees C), a ladder of protein bands in approximately 55 kDa increments were detected above the monomeric p55 band. These discrete bands visualized as a ladder are likely the disulfide-linked L1 oligomers. In addition to the gel running conditions, an increase in pH, temperature, or SDS concentration during sample treatment was also shown to significantly reduce the amount of detectable oligomers, further corroborating the labile nature of these oligomers. Altogether, the results also implicate the redox-responsive nature of the HPV capsid comprising of >95% L1 protein. Molecular basis of the facile disulfide bond inter-change is discussed. This electrophoretic technique for trapping the disulfide-linked oligomers may be employed to detect the oligomeric status of other protein aggregates or assembled particles. PMID- 15894388 TI - Method for detection and quantitation of fathead minnow vitellogenin (Vtg) by liquid chromatography and matrix-assisted laser desorption/ionization mass spectrometry. AB - Vitellogenin (Vtg) is a well-recognized biomarker of estrogen exposure in many species, particularly fish. This large protein shares a high degree of sequence homology across a large number of species. Quantitative measurement is currently done using antibody-based assays. These assays frequently require purification of Vtg and antibody production from each species because there is poor cross reactivity between antibodies for different fish. Therefore, complementary methods of measuring Vtg are desirable. Mass spectrometric (MS) analysis coupled to database searching offers the promise of a general method for protein identification. In this study, we used the well-characterized Vtg from rainbow trout (Oncorhynchus mykiss) to evaluate the analytical parameters for matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis of the intact protein. The low sensitivity observed for the intact protein demonstrated that a proteolytic digestion would be necessary for MALDI-MS identification of Vtg. An analytical scale high performance liquid chromatography (HPLC) separation combined with MALDI-MS was used to measure and confirm the identity of Vtg from the plasma of an important species for regulatory agencies, fathead minnow (Pimephales promelas). The small volume requirement of this method (<10 microL) was found to be compatible with the plasma volume obtained from a few minnows. Importantly, the HPLC-MALDI-MS measurement of fathead minnow Vtg abundance after exposure to estradiol was similar to previously obtained enzyme linked immunosorbant assays (ELISA) data. PMID- 15894389 TI - Identification and characterisation of two runx2 homologues in zebrafish with different expression patterns. AB - Genome and gene duplications are considered to be the impetus to generate new genes, as the presence of multiple copies of a gene allows for paralogues to adopt novel function. After at least two rounds of genome/gene duplication, the Runt gene family consists of three members in vertebrates, instead of one in invertebrates. One of the family members, Runx2, plays a key role in the development of bone, a tissue that first occurs in vertebrates. The family has thus gained new gene function in the course of evolution. Two Runx2 genes were cloned in the vertebrate model system the zebrafish (Danio rerio). The expression patterns of the two genes differ and their kinetics differ up to four fold. In addition, splice forms exist that are novel when compared with mammals. Together, these findings comprise opportunities for selection and retention of the paralogues towards divergent and possibly new function. PMID- 15894390 TI - Multiple proteins are involved in the protein-DNA complex in the proximal promoter of the human alpha1(III) collagen gene (COL3A1). AB - We have characterized the proximal promoter of the human alpha1(III) collagen gene (COL3A1). Transient transfection assays using a series of chimeric constructs linked to the luciferase gene indicated that the segment from -96 to 34 is necessary to activate transcription. Electrophoretic mobility shift assays (EMSAs) showed that the multiple proteins form the DNA-protein complex in different combinations depending on the cell types. A competition assay using mutant oligonucleotides showed that the sequence 5'-GCTCTCATATTTCAGAA-3' (-79 to 63 bp) is critical for DNA-protein complex formation. This sequence is contained in the B element of mouse alpha1(III) collagen gene (Col3a1) reported by Ruteshouse and de Crombrugghe (J. Biol. Chem., 1993). In the rhabdomyosarcoma cell line, A204, at least two proteins of 92-118 kDa and 40-52 kDa are involved in the DNA-protein complex bound to this motif. PMID- 15894391 TI - Human myotonic dystrophy protein kinase effect in S. cerevisiae. AB - Human myotonic dystrophy protein kinase (DMPK), the product of the myotonic dystrophy (DM) locus, is a member of a novel class of multidomain serine threonine protein kinases, which interacts with members of the Rho family of small GTPases. DMPK has been shown to affect the cell growth, size and shape in different organisms, from fission yeast to man, but its physiological role is still unclear. We examined the effect of the overexpression of two forms of human DMPK, full-length (DMFL) and a C-terminal truncated form (DMT) on the growth and cell morphology of S. cerevisiae, which possesses a DMPK homologous gene (CBK1) important for polarized growth and cell division. We report that the overexpression of either forms of human DMPK did not complement the CBK1 function in the haploid strain WR208-1a, deleted for CBK1. The truncated form, but not the full length one, slowed down growth rate and induced elongation of the haploid wild type strain CBK1. Similar results were obtained in the diploid wild type strain RS112 of S. cerevisiae where also the full-length form was effective. These effects were abolished when either DMFL or DMT were mutated in the ATP binding site (K100R mutation), suggesting that the kinase activity of DMPK is required. Interestingly, DMPK localization in yeast is similar to that of Cbk1 protein suggesting that it might affect a pathway, which regulates cell morphogenesis and progression through cell cycle, possibly involving CBK1. PMID- 15894392 TI - On the surveillance for animal diseases in small herds. AB - Small herds may present a problem in surveillance for infectious animal diseases because typical levels of a within-herd design prevalence are not directly applicable. We suggest a definition of small herds as those smaller than 2/(within-herd design prevalence) on the basis that such herds would be expected to have less than two (i.e. only one) infected animals. Consequently, the probability of detecting small herds cannot be improved by choosing a larger sample size within the herd. We derive necessary sample sizes of herds and the probability ("confidence") of detecting disease within a stratum of small herds, given the among-herd design prevalence and test diagnostic sensitivity. Both a binomial model and a Poisson model can be used to establish the confidence for a given sample size of herds (and vice versa). The results of a simulation study suggest that the Poisson model provides more conservative (lower) estimates of the confidence for a given sample size and should therefore be preferred. PMID- 15894393 TI - Buccal penetration enhancers--how do they really work? AB - Certain agents that increase drug delivery through the skin, including surfactants, bile salts, and fatty acids, have been shown to exert a similar effect on the buccal mucosa. These agents enhance skin permeability by interacting with and disrupting the ordered intercellular lipid lamellae within the keratinized stratum corneum, and it has been assumed that a similar mechanism of action occurs in the nonkeratinized buccal mucosa. However, the chemical and structural nature of the lipids present within the intercellular regions of the buccal mucosa is quite different to that found within the stratum corneum, and so extrapolation of results between these two tissues may be misleading. To assume that the mechanism of action of buccal penetration enhancers is based on the disruption of intercellular lipids may be erroneous, and may result in the inappropriate prediction that certain skin penetration enhancers will similarly enhance drug delivery through the buccal mucosa. The data available in the literature suggest that agents that enhance buccal penetration exert their effect by a mechanism other than by disruption of intercellular lipids. Rather, buccal penetration enhancement appears to result from agents being able to (a) increase the partitioning of drugs into the buccal epithelium, (b) extract (and not disrupt) intercellular lipids, (c) interact with epithelial protein domains, and/or (d) increase the retention of drugs at the buccal mucosal surface. The purpose of this review is to identify the major differences in the structural and chemical nature of the permeability barriers between the buccal mucosa and skin, to clarify the mechanisms of action of buccal penetration enhancers, and to identify the limitations of certain models that are used to assess the effect of buccal penetration enhancers. PMID- 15894394 TI - Synthesis, characterization, and tumor selectivity of a polyphosphazene platinum(II) conjugate. AB - A new amphiphilic poly(organophosphazene) was synthesized by stepwise nucleophilic substitutions with a hydrophilic methoxy poly(ethylene glycol) with an average molecular weight of 350 (MPEG350) and a hydrophobic glycyl-L-glutamate as side groups, and then an antitumor (dach)platinum(II) (dach: trans-(+/-)-1,2 diaminocyclohexane) moiety was conjugated to the polymer using the dipeptide as a spacer. This polymeric platinum conjugate was found to be accumulated in the tumor tissue to a remarkably greater extent than in the normal tissue (tumor/tissue ratio >4), probably due to the excellent EPR effect and the long circulating properties of the polymer conjugate (t1/2beta=6.2 h and AUC=4020 nmol h/ml) compared with carboplatin (t1/2beta=0.42 h and AUC=120 nmol h/ml). The polymer conjugate also exhibited high in vitro cytotoxicity comparable to cisplatin against several human tumor cells tested. PMID- 15894395 TI - Effect of electroporation on the electroosmosis across hairless mouse skin in vitro. AB - The effect of electroporation on the iontophoresis-produced electroosmosis across the skin was evaluated by measuring the permeability of hairless mouse skin, to mannitol, a non-electrolyte, in vitro. Immediately after electroporation by squared pulses (10 times/s) at 100, 150 or 200 V for 1 ms, anodal iontophoretic permeations were determined at 0.4 mA/cm2 for 4 h. The observed iontophoretic permeability of mannitol was higher with electroporation pretreatment than without pretreatment. The enhanced flux of mannitol induced by electroporation, however, was due to increased passive diffusion. The contribution of convective or osmotic flow caused by anodal iontophoresis on skin permeation of mannitol was decreased by the pretreatment. In addition, osmotic flow was decreased with an increase in the applied voltage for electroporation. In contrast, mannitol flux during cathodal iontophoresis at 0.4 mA/cm2 after 150 or 200 V electroporation was higher than without electroporation as well as anodal iontophoresis, but cathodal iontophoretic flux after electroporation was lower than without iontophoresis. The neutral high-molecular compound dextran rhodamine B was also used as a second model. Anodal iontophoresis alone did not increase skin permeability of the compound. However, electroporation pretreatment before anodal iontophoresis enhanced the skin permeation of dextran rhodamine B, which was due to increased osmotic flow induced by this combination. These results suggest that electroporation decreases the electroosmosis produced by iontophoresis, and that electroporation increases skin permeability to neutral low and high model compounds (mannitol and dextran rhodamine B) probably due to an enlarged permeation pathway. Thus, electroporation affects osmotic flow from the anode to cathode during iontophoresis. Therefore, one has to pay attention to the change in electroosmosis produced by iontophoresis for the combined use of electroporation and iontophoresis to attain a high skin-penetration enhancing effect. PMID- 15894396 TI - Aspirin sensitivity of platelet aggregation in diabetes mellitus. AB - Although aspirin is cardioprotective in high-risk populations, many with diabetes mellitus (DM) are unresponsive to these benefits. We questioned whether cardiovascular unresponsiveness might be demonstrated by lack of aspirin sensitivity to in vitro platelet functions especially in subjects with poorly controlled diabetes. Six women and 4 men (48+/-8 years [mean+/-S.D.]), selected for poor control (glycohemoglobin 11.9+/-2.2%) and 10 sex-age (+/-5 years) matched controls received 81 mg aspirin daily. There was a 2-week washout from aspirin and related drugs. After the aspirin dose on day-7, blood for platelet aggregation assays, and 24-h urine for 2,3 dinor thromboxane B2 (TxB2) and 2,3 dinor 6-keto (PGF1alpha) were obtained. Aspirin sensitivity was defined as inhibition (i.e., lower than expected) platelet aggregation after exposure to an agonist. Those with diabetes and controls were sensitive to aspirin inhibition of platelet aggregation induced by 1.6 mM arachidonic acid (9.5+/-3.9% versus 9.1+/ 3.1%, normal range 40-100%) and by 0.83 microg/mL collagen (17.4+/-13.9% versus 13.2+/-9.3%, normal range 60-93%), respectively. Aspirin sensitivity to 2 microM ADP was present in five with diabetes and five controls. Urinary prostaglandin metabolites were suppressed below reference ranges, without differences between those with DM or controls for TxB2 (350+/-149 pg/mg versus 348+/-93 pg/mg creatinine) and PGF1alpha (255+/-104 pg/mg versus 222+/-88 pg/mg creatinine). In conclusion, in poorly controlled diabetes, there was no differential lack of aspirin sensitivity to platelet aggregation, or lack of aspirin suppression of urinary TxB2 or PGF1alpha, compared with controls on aspirin. Despite suppression of urinary prostaglandin metabolites, aspirin resistance was most apparent to ADP mediated platelet aggregation. It is not known what level of inhibition of in vitro tests is necessary for the cardioprotective benefits of aspirin in diabetes mellitus. Thus, the lack of aspirin protection in diabetes may be due to undefined aspects of platelet function. PMID- 15894398 TI - Proteomics in the study of liver pathology. PMID- 15894397 TI - Prediction of hepatic fibrosis in HIV/HCV co-infected patients using serum fibrosis markers: the SHASTA index. AB - BACKGROUND/AIMS: To examine if serum fibrosis biomarkers could accurately identify the stage of liver disease amongst hepatitis C (HCV) and HIV co-infected patients. METHODS: One hundred and thirty seven HIV/HCV co-infected persons were randomly selected from the Johns Hopkins HIV Clinic cohort. Ninety five had complete testing for fibrosis markers in sera collected at the time of liver biopsy. Biopsies were scored according to Ishak modified histological activity index (F0 no fibrosis to F6 cirrhosis). Fibrosis was evaluated against alanine aminotransferase (ALT), aspartate aminotransferase (AST), AST to platelet ratio (APRI), albumin, total bilirubin, hyaluronic acid (HA) and YKL-40. RESULTS: Sixty nine (73%) had no or minimal portal fibrosis (F0-2) and were compared with remaining subjects (F3-6). Fibrosis scores > or =F3 were found 27 times more often in persons with HA levels >86 ng/ml and 5.5 times more often in persons with HA levels 41-86 ng/ml. Less substantial associations were detected with levels of albumin <3.5 g/dl (OR 4.85) and AST >60 iu (OR 5.91). All 35 subjects who had favorable results of HA, albumin, and AST had minimal fibrosis (F0-2). CONCLUSIONS: Amongst HIV/HCV co-infected patients, serum testing for HA, albumin, and AST (SHASTA Index) was able to accurately stage mild and advanced fibrosis. PMID- 15894400 TI - Pediatric gallstone disease in familial hypobetalipoproteinemia. AB - Familial hypobetalipoproteinemia (FHBL) is an monogenic co-dominant disorder characterized by reduced plasma levels of cholesterol, low density lipoproteins (LDL) and apolipoprotein B (apoB) often associated with non-alcoholic fatty liver disease (NAFLD). It has been suggested that FHBL might predispose to gallstone disease (GD). We report a hypocholesterolemic 10 year old girl with obstructive jaundice due to cholesterol stones in gallbladder and common bile duct which required cholecistectomy. The analysis of patient's plasma lipoproteins revealed a marked reduction of LDL and apoB, a lipid profile consistent with the clinical diagnosis of heterozygous FHBL. The same profile was found in her mother who had severe NAFLD. The analysis of apoB gene, the main candidate gene in FHBL, revealed that the patient and her mother were heterozygotes for a novel nonsense mutation (Y1220X) predicted to cause the formation of a short truncated apoB (apoB-26.87) not secreted into the plasma. The presence of cholesterol stones could result from increased biliary cholesterol secretion as a compensatory mechanism for the reduced capacity of the liver to export cholesterol incorporated into apoB-containing lipoproteins. FHBL should be considered as a possible predisposing factor for cholesterol gallstones in children (190). PMID- 15894401 TI - Microfocal X-ray computed tomography post-processing operations for optimizing reconstruction volumes of stented arteries during 3D computational fluid dynamics modeling. AB - Restenosis caused by neointimal hyperplasia (NH) remains an important clinical problem after stent implantation. Restenosis varies with stent geometry, and idealized computational fluid dynamics (CFD) models have indicated that geometric properties of the implanted stent may differentially influence NH. However, 3D studies capturing the in vivo flow domain within stented vessels have not been conducted at a resolution sufficient to detect subtle alterations in vascular geometry caused by the stent and the subsequent temporal development of NH. We present the details and limitations of a series of post-processing operations used in conjunction with microfocal X-ray CT imaging and reconstruction to generate geometrically accurate flow domains within the localized region of a stent several weeks after implantation. Microfocal X-ray CT reconstruction volumes were subjected to an automated program to perform arterial thresholding, spatial orientation, and surface smoothing of stented and unstented rabbit iliac arteries several weeks after antegrade implantation. A transfer function was obtained for the current post-processing methodology containing reconstructed 16 mm stents implanted into rabbit iliac arteries for up to 21 days after implantation and resolved at circumferential and axial resolutions of 32 and 50 microm, respectively. The results indicate that the techniques presented are sufficient to resolve distributions of WSS with 80% accuracy in segments containing 16 surface perturbations over a 16 mm stented region. These methods will be used to test the hypothesis that reductions in normalized wall shear stress (WSS) and increases in the spatial disparity of WSS immediately after stent implantation may spatially correlate with the temporal development of NH within the stented region. PMID- 15894402 TI - CAT 53: a protein phosphatase 1 nuclear targeting subunit encoded in the MHC Class I region strongly expressed in regions of the brain involved in memory, learning, and Alzheimer's disease. AB - We identified CAT 53 by cDNA hybridization selection as an expressed sequence tag (EST), located in the vicinity of HLA-C and designated as CAT (for HLA-C associated transcript) 53. CAT 53 encodes a protein described by others and commonly known as phosphatase 1 nuclear targeting subunit (PNUTS). PNUTS is a potent inhibitor of nuclear serine/threonine protein phosphatase 1 (PP1). We present the genomic organization of CAT 53, localize specific sites of mRNA transcription in thin sections of mouse brain by in-situ hybridization, and perform a structural analysis of the peptide domains. We also characterize the protein expression pattern for PNUTS by Western blotting and immunohistochemistry with PNUTS antibody in Alzheimer's disease (AD) brains and age-matched control brains. In-situ hybridization and immunohistochemistry analysis of human and mouse brain show high CAT 53 expression in the olfactory cortex, piriform cortex, and hippocampus. Very high expression of CAT 53 was found mainly in the hippocampus, frontal, and entorhinal cortex of control brains and in the neurofibrillary tangles of AD brain. In the hippocampus, CAT 53 is expressed in CA1 and CA3 cell layers and in the dentate gyrus. The hippocampus is known to play a fundamental role in learning and episodic memories and has been implicated in a number of neurological and psychiatric disorders, including AD, epilepsy, and schizophrenia. Our findings suggest that PNUTS, encoded by CAT 53 on 6p21.3, may have a role in the progression of AD. PMID- 15894403 TI - Prognostic factors in mesothelioma. AB - Prognostic factors can help clinicians and patients when deciding a treatment plan. Patients in the best prognostic groups can be considered for more intensive or experimental therapy. Alternatively, patients in the best prognostic groups might prefer a period of observation prior to commencement of therapy. For patients with mesothelioma prognostic factors are potentially especially important because of the lack of a widely applicable anatomical staging system. Both the International Mesothelioma Interest Group (IMIG) and Brigham staging systems are of limited relevance to patients not undergoing radical debulking surgery. Radiological prediction of IMIG or Brigham stage is of little value. Review of the best-known prognostic scoring systems from the EORTC and CALGB has shown that the most important predictors of poor prognosis are: poor performance status; non-epithelioid histology; male gender; low hemoglobin; high platelet count; high white blood cell count; and high lactate dehydrogenase (LDH). The EORTC model was validated at St Bartholomew's Hospital in a group of 145 patients treated in sequential phase II chemotherapy trials. For 70 patients treated with vinorelbine, those having the best EORTC prognosis had a median survival of 19.2 months [95% C.I.=14.7-23.7] compared to 9.9 months [95% C.I.=8.5-11.3] for those in the worst group. The suggestion is that all clinical and biological factors relevant to prognosis should be recorded prospectively in mesothelioma patients selected for clinical trials. PMID- 15894404 TI - Functional foods: health claim-food product compatibility and the impact of health claim framing on consumer evaluation. AB - Two studies are reported, which aim to strengthen the scientific underpinning of strategic decisions regarding functional food development, as to (1) which health benefits to claim, (2) with which product (category), and (3) in which communication format. The first exploratory study is a secondary analysis of 10 different health claims systematically combined with 10 different food carriers to evaluate their combined suitability for functional food positioning. The results show that consumers tend to prefer functional food concepts that primarily communicate disease-related health benefits in carriers with a healthy image or health positioning history. Study 2 examines health claim format and systematically varies the way in which specific health benefits are being communicated to the consumer. Two physiologically oriented claims (heart disease and osteoporosis) and two psychologically oriented food claims (stress and lack of energy) are expressed in enhanced function format versus disease risk reduction format. Also, it includes the individual difference variable of 'regulatory focus' and the health status of the respondent to explore how these factors impact health claim evaluation. The results show that consumer evaluations primarily differ to the extent that health claims are personally relevant in addressing an experienced disease state. Framing is important, but its effect differs by health benefit. No strong effects for consumers' regulatory focus were found. Underlying mechanisms of these effects and their implications for the development of functional foods are discussed. PMID- 15894405 TI - Correlation between the activities of alpha-helical antimicrobial peptides and hydrophobicities represented as RP HPLC retention times. AB - PTP7 is a 13-amino acid residue peptide designed from gaegurin 6, an antimicrobial peptide isolated from skin secretions of Rana rugosa. In order to examine the effect of hydrophobicity on antimicrobial activity, a series of PTP7 derivatives were constructed and analyzed the activity against bacteria and artificial membrane. We found that the mean hydrophobicity by simple summation of hydrophobicity of each constituent amino acid did not necessarily describe the hydrophobic property of antimicrobial peptides. The mean hydrophobicity did not show close correlation with the observed hydrophobicity by measuring reverse phase high performance liquid chromatography (RP HPLC) retention time. The observed hydrophobicity represented as RP HPLC retention time correlated well with the activity against artificial membrane and Gram positive bacterial species, such as Staphylococcus aureus, Staphylococcus epidermidis, and Micrococcus luteus, rather than mean hydrophobicity. However, antimicrobial activity against Gram negative bacteria, such as Escherichia coli, did not show correlation with RP HPLC retention time. These data indicate that the RP HPLC retention time should be exploited rather than the mean hydrophobicity in the analysis of the relationship between hydrophobicity and antimicrobial activity. PMID- 15894406 TI - Feeding, body weight, and sensitivity to non-ingestive reward stimuli during and after 12-day continuous central infusions of melanocortin receptor ligands. AB - The brain melanocortin system mediates downstream effects of hypothalamic leptin and insulin signaling. Yet, there have been few studies of chronic intracerebroventricular (i.c.v.) melanocortin receptor (MCR) agonist or antagonist infusion. Although there is evidence of interaction between melanocortin and dopamine (DA) systems, effects of chronic MCR ligand infusion on behavioral sensitivity to non-ingestive reward stimuli have not been investigated. The objective of this study was to investigate effects of chronic i.c.v. infusion of the MCR agonist, MTII, and the MCR antagonist, SHU9119, on food intake, body weight, and sensitivity to rewarding lateral hypothalamic electrical stimulation (LHSS) and the reward-potentiating (i.e., threshold lowering) effect of D-amphetamine. The MCR antagonist, SHU9119 (0.02 microg/h) produced sustained hyperphagia and weight gain during the 12-day infusion period, followed by compensatory hypophagia and an arrest of body weight gain during the 24-day post-infusion period. At no point during the experiment was sensitivity to LHSS or D-amphetamine (0.25mg/kg, i.p.) altered. The MCR agonist, MTII (0.02 microg/h) produced a brief hypophagia (3 days) followed by a return to control levels of daily intake, but with body weight remaining at a reduced level throughout the 12-day infusion period. This was followed by compensatory hyperphagia and weight gain during the 24-day post-infusion period. There was no change in sensitivity to non-ingestive reward stimuli during the infusion of MTII. However, sensitivity to D-amphetamine was increased during the 24-day post infusion period. It therefore seems that changes in ingestive behavior that occur during chronic MCR ligand infusion may not affect the response to non-ingestive reward stimuli. However, it is possible that the drive to re-feed and restore body weight following MCR agonist treatment includes neuroadaptations that enhance the incentive effects of drug stimuli. PMID- 15894407 TI - White matter mapping in Alzheimer's disease: A neuropathological study. AB - White matter disease (WMD) with pervasive non-focal subtotal tissue loss is frequently seen in Alzheimer's disease (AD) upon neuropathological examination. Although WMD has varying effects on AD symptoms, accurate clinical detection is difficult due partly to scarcity of correlative structural imaging and histopathological studies. Neuropathological studies of WMD severity and distribution have been conducted earlier using semi-quantitative methods. A technique for quantifying WMD objectively in large white matter areas, based on optical density (OD) measurements on images of scanned whole-brain sections, was developed and was validated using conventional microscopic assessment. Altogether, 16 AD cases with concomitant WMD (AD-WMD) and 9 cases of AD without WMD (AD-only) were analysed. The OD values correlated significantly with the neuropathological severity of WMD and were significantly lower in AD-WMD than in AD-only in frontal, frontoparietal, temporal and parietal white matter but not in the occipital white matter, the frontal OD difference being greatest. Useful baseline information on WMD distribution in AD to relate to in vivo imaging results was obtained. PMID- 15894408 TI - Bosentan preserves endothelial function in mice overexpressing APP. AB - This study was designed to test the hypothesis that Alzheimer's disease (AD) is associated with endothelial dysfunction and that chronic endothelin-1 antagonism preserves endothelial function in mice overexpressing the AD amyloid precursor protein (APP). Three groups of mice were studied: C57BL/6 (normal control, n = 6), transgenic mice overexpressing APP (Tg2576, n = 5), and Tg2576 mice fed Bosentan (100 mg/(kg day)(-1)), a combined endothelin A and B receptor antagonist, for 4 months (Tg2576+Bosentan, n = 5). Mice were sacrificed at the age of 7 months. In vitro, the endothelium-dependent aortic vasorelaxation was significantly attenuated in Tg2576 mice as compared to C57BL/6 and Tg2576+Bosentan mice. In contrast, Tg2576+Bosentan and C57BL/6 mice showed similar endothelium-dependent aortic vasorelaxation. Similarly, endothelium dependent carotid vasorelaxation was significantly attenuated in Tg2576 mice compared to C57BL/6 and Tg2576+Bosentan mice. There was no difference between the three groups in the response to nitroprusside. The current study demonstrates the presence of endothelial dysfunction in both carotid and aortic arteries in mice overexpressing APP and suggests a pathophysiological role for the endogenous endothelin system in AD. PMID- 15894409 TI - Beta-amyloid deposition and Alzheimer's type changes induced by Borrelia spirochetes. AB - The pathological hallmarks of Alzheimer's disease (AD) consist of beta-amyloid plaques and neurofibrillary tangles in affected brain areas. The processes, which drive this host reaction are unknown. To determine whether an analogous host reaction to that occurring in AD could be induced by infectious agents, we exposed mammalian glial and neuronal cells in vitro to Borrelia burgdorferi spirochetes and to the inflammatory bacterial lipopolysaccharide (LPS). Morphological changes analogous to the amyloid deposits of AD brain were observed following 2-8 weeks of exposure to the spirochetes. Increased levels of beta amyloid precursor protein (AbetaPP) and hyperphosphorylated tau were also detected by Western blots of extracts of cultured cells that had been treated with spirochetes or LPS. These observations indicate that, by exposure to bacteria or to their toxic products, host responses similar in nature to those observed in AD may be induced. PMID- 15894410 TI - Differences in regional brain atrophy in genetic forms of Alzheimer's disease. AB - Multiple degenerative hallmarks characterize Alzheimer's disease: insoluble protein deposition, neuronal loss and cortical atrophy. Atrophy begins in the medial temporal lobe and becomes global by end stage. In a small proportion of cases, these tissue changes are caused by mutations in three known genes. These cases are affected earlier in life and have more abundant protein deposition, which may indicate greater tissue atrophy and degeneration. This issue remains unresolved. Grey matter atrophy in different cortical regions was determined in genetic cases of Alzheimer's disease (N = 13) and compared to sporadic cases (N = 13) and non-diseased controls (N = 23). Genetic mutations were found to influence the degree and regional pattern of atrophy. The majority of cases had greater medial temporal atrophy than sporadic disease, suggesting that abnormalities affecting Abeta metabolism selectively increase hippocampal degeneration. Cases with mutations in presenilin-1 demonstrated additional increased frontotemporal atrophy. This effect may be due to the influence of presenilin-1 on tau phosphorylation and metabolism. These differences may explain the earlier onset ages in these different forms of Alzheimer's disease. PMID- 15894411 TI - Self-directed learning: implications and limitations for undergraduate nursing education. AB - Self-directed learning (SDL) is an educational concept that has received increasing attention in recent years, particularly in the context of higher education. Whilst the benefits of SDL have been espoused by a number of adult education theorists (Brookfield, S., 1986. Understanding and Facilitating Adult Learning. Jossey-Bass, San Francisco; Houle, C., 1984. Patterns of Learning: New Perspectives on Life-Span Education. Jossey-Bass, San Francisco; Knowles, M., 1998. The Adult Leaner: A Neglected Species, fifth ed., Gulf, Houston; Tough, A., 1979. The Adults Learning Project: A Fresh Approach to Theory and Practice in Adult Learning. Ontario Institute for Studies in Education, Toronto), its introduction into curricula has not always been successful (Nolan, J., Nolan, M., 1997a. Self-directed and student-centred learning in nurse education: 1. British Journal of Nursing 6 (1), 51-55; Nolan, J., Nolan, M., 1997b. Self-directed and student-centred learning in nurse education: 2. British Journal of Nursing 6 (2), 103-107; Slevin, O., Lavery, M., 1991. Self-directed learning and student supervision. Nurse Education Today 11, 368-377). The indiscriminate application of SDL principles and poorly prepared teachers and/or students has at times meant that the introduction of SDL has been resented rather than welcomed (Iwasiw, C., 1987. The role of the teacher in self-directed learning. Nurse Education Today 7, 222-227; Turunen, H., Taskinen, H., Voutilainen, U., Tossavainen, K., Sinkkonen, S., 1997. Nursing and social work students' initial orientation towards their studies. Nurse Education Today 17, 67-71). This paper clarifies and explores these issues by: (a) examining the origins of SDL; (b) discussing the relevance of self-directed learning to Knowles' theory of adult learning and contemporary educational practices such as enquiry based learning and problem based learning; and (c) highlighting the implications and limitations of SDL with regard to adult education in general, and undergraduate nursing education in particular. PMID- 15894412 TI - Uptake of heavy metals and As by Brassica juncea grown in a contaminated soil in Aznalcollar (Spain): the effect of soil amendments. AB - Two crops of Brassica juncea (L.) Czern. were grown in a field experiment, at the site affected by the toxic spillage of acidic, metal-rich waste in Aznalcollar (Seville, Spain), to study its metal accumulation and the feasibility of its use for metal phytoextraction. The effects of organic soil amendments (cow manure and mature compost) and lime on biomass production and plant survival were also assessed; plots without organic amendment and without lime were used as controls. Plots, with or without organic amendment, having pH < 5 were limed for the second crop. Soil acidification conditioned plant growth and metal accumulation. The addition of lime and the organic amendments achieved higher plant biomass production, although effects concerning metal bioavailability and accumulation were masked somewhat by pH variability with time and between and within plots. Tissue metal concentrations of B. juncea were elevated for Zn, Cu and Pb, especially in leaves of plants from plots with low pH values (maxima of 2029, 71 and 55 microg g(-1), respectively). The total uptake of heavy metals in the plants was relatively low, emphasising the problems faced when attempting to employ phytoextraction for clean-up of pluri-contaminated sites. PMID- 15894413 TI - Depauperate macroinvertebrates in a mine affected stream: clean water may be the key to recovery. AB - Acid mine drainage (AMD) is frequently linked with changes in macroinvertebrate assemblages, but the relative contribution of water and sediment to toxicity is equivocal. We have shown that the macroinvertebrate fauna of Neubecks Ck, a mine impacted stream in New South Wales, Australia, was much poorer than in two reference streams. Multivariate RELATE analyses indicated that the patterns in the biological data were more strongly correlated with the concentrations of common metals in the surface water than the pore water of these streams. From this we hypothesised that the water was more toxic to the biota than the sediment and we tested this hypothesis with a sediment transplant experiment. Sediment from Neubecks Ck that was placed in reference streams retained high concentrations of metals throughout the experiment, yet supported a macroinvertebrate assemblage similar to that in the reference streams. Sediment from the reference streams that was placed in Neubecks Ck supported few, if any, animals. This indicates that water in Neubecks Ck is toxic to biota, but that sediment is able to support aquatic biota in clean water. Therefore, remediation should focus on improving water quality rather than sediment quality. PMID- 15894414 TI - Phytoremediation in the tropics--influence of heavy crude oil on root morphological characteristics of graminoids. AB - When studying species for phytoremediation of petroleum-contaminated soils, one of the main traits is the root zone where enhanced petroleum degradation takes place. Root morphological characteristics of three tropical graminoids were studied. Specific root length (SRL), surface area, volume and average root diameter (ARD) of plants grown in crude oil-contaminated and uncontaminated soil were compared. Brachiaria brizantha and Cyperus aggregatus showed coarser roots in polluted soil compared to the control as expressed in an increased ARD. B. brizantha had a significantly larger specific root surface area in contaminated soil. Additionally, a shift of SRL and surface area per diameter class towards higher diameters was found. Oil contamination also caused a significantly smaller SRL and surface area in the finest diameter class of C. aggregatus. The root structure of Eleusine indica was not significantly affected by crude oil. Higher specific root surface area was related to higher degradation of petroleum hydrocarbons found in previous studies. PMID- 15894415 TI - Tree age dependence and within-canopy variation of leaf gas exchange and antioxidative defence in Fagus sylvatica under experimental free-air ozone exposure. AB - We characterized leaf gas exchange and antioxidative defence of two-year-old seedlings and 60-year-old trees of Fagus sylvatica exposed to ambient (1 x O3) or two-fold ambient (2 x O3) O3 concentrations (maximum of 150 ppb) in a free-air canopy exposure system throughout the growing season. Decline in photosynthesis from sun-exposed to shaded conditions was more pronounced in adult than juvenile trees. Seedling leaves and leaves in the sun-exposed canopy had higher stomatal conductance and higher internal CO2 concentrations relative to leaves of adult trees and leaves in shaded conditions. There was a weak overall depression of photosynthesis in the 2 x O3 variants across age classes and canopy positions. Pigment and tocopherol concentrations of leaves were significantly affected by canopy position and tree age, whereas differences between 1 x O3 and 2 x O3 regimes were not observed. Glutathione concentrations were significantly increased under 2 x O3 across both age classes and canopy levels. Seedlings differed from adult trees in relevant physiological and biochemical traits in ozone response. The water-soluble antioxidative systems responded most sensitively to 2 x O3 without regard of tree age or canopy position. PMID- 15894416 TI - Does NHS Direct empower patients? AB - NHS Direct is a 24h telephone helpline established in England and Wales, UK to offer advice and information for people about health, illness and the National Health Service (NHS) so that they are better able to care for themselves and their families. In 2001/2002 we undertook in-depth home interviews with 60 users of the service in two NHS Direct sites in England. In this paper we consider the extent to which NHS Direct facilitates patient empowerment in terms of helping people to be in control of their health and health care interactions. Our research suggests that NHS Direct facilitates patient empowerment by enabling patients to self care and to access health advice and services. It is also seen to offer the prerequisites for empowerment perceived to be lacking in the wider NHS, including time, respect, listening, support, and information. The service also functions by offering an alternative contact point for people seeking to avoid being labelled 'time wasters' by other busy health care providers. In the context of a wider health service which appears to problematise individuals' ability to make decisions about the appropriateness of seeking health care, NHS Direct legitimises help-seeking actions. Empowerment in the context of NHS Direct has been associated with self care as a way of reducing 'unnecessary' demand on health services. However, health professional and patient perspectives on what is considered necessary demand differ, and in certain contexts, patient empowerment may involve service use as well as self care. Further, our data reveal the context-dependent nature of a concept like empowerment. For example, when people are ill, in pain, or anxious about a loved one, they may value being cared for more than being empowered. Our research suggests that, in addition to its other functions, NHS Direct is also valued as contributing to a sense of being cared for. PMID- 15894417 TI - The association between p53 expression, stage and histological features in endometrial cancer. AB - OBJECTIVE: Alterations of the p53 gene have been widely suggested to be relevant to the development of endometrial carcinoma. However, contradictory results have been reported when immunohistochemical determination of p53 expression has been correlated with stage and histological features of the tumours. STUDY DESIGN: Pathology findings were reviewed and p53 immunoperoxidase staining was performed in 240 cases of endometrial carcinoma. RESULTS: Uterine papillary serous adenocarcinomas showed significantly higher p53 overexpression than uterine endometrioid adenocarcinomas (100.0% versus 61.0%, p<0.005). p53 overexpression was significantly higher in the secretory variant (85.7%) than in the typical endometrioid carcinoma (60.0%) (p<0.05). p53 expression did not differ between early (stage I) and advanced (stage II-IV) carcinomas. Likewise, no difference was observed in p53 expression among different architectural grades. The incidence of metastasis to lymph nodes was similar in p53 positive (13.7%) and in p53 negative tumours (12.5%). CONCLUSION: In the present series, p53 immunostaining did not differ between cases with different FIGO stages or histologic characteristics of the tumours. No simple relationship exists between the immunohistochemical determination of p53 expression and the biological aggressiveness of endometrial carcinomas. PMID- 15894418 TI - Increased levels of macrophage migration inhibitory factor (MIF) in preeclampsia. AB - OBJECTIVE: MIF is a proinflammatory cytokine involved in reproduction. Systemic activation of maternal inflammatory cell responses may play an important role in the pathogenesis of preeclampsia (PE). We hypothesized that MIF could be involved in preeclampsia. STUDY DESIGN: Concentration of immunoreactive MIF was assayed by enzyme-linked immunoassorbent assay (ELISA) in maternal serum samples obtained from 41 term control pregnancies and 21 severe preeclamptic pregnancies (14 delivered before and 7 at or after 34 weeks). RESULTS: MIF serum levels were significantly higher in preeclamptic pregnancies (median 12.74 ng/ml) than in control group (median 5.3n g/ml) p = 0.001. MIF concentration was significantly higher when delivery occurred <34 weeks (median 17.80 ng/ml; range 2.80-80.20) than in the group delivered > or = 34 weeks (median 6.16 ng/ml; range 1.62-23.65) p = 0.037. CONCLUSIONS: High maternal serum levels MIF in pregnancies complicated by severe preeclampsia strongly support the role of inflammation in the pathogenesis of this disease. PMID- 15894419 TI - A study of the perturbation effects of the local anesthetic procaine on human erythrocyte and model membranes and of modifications of the sodium transport in toad skin. AB - The interaction of the local anesthetic procaine with human erythrocytes, isolated unsealed human erythrocyte membranes (IUM), isolated toad skins, and molecular models is described. The latter consisted of phospholipid multilayers built-up of dimyristoylphosphatidylcholine (DMPC) and of dimyristoylphosphatidylethanolamine (DMPE), representatives of phospholipid classes located in the outer and inner monolayers of the human erythrocyte membrane, respectively. Optical and scanning electron microscopy of human erythrocytes revealed that procaine induced the formation of stomatocytes. Experiments performed on IUM at 37 degrees C by fluorescence spectroscopy showed that procaine interacted with the phospholipid bilayer polar groups but not with the hydrophobic acyl chains. X-ray diffraction indicated that procaine perturbed DMPC structure to a higher extent when compared with DMPE, its polar head region being more affected. Electrophysiological measurements disclosed a significant decrease in the potential difference (PD) and in the short-circuit current (Isc) after the application of procaine to isolated toad skin, reflecting inhibition of active ion transport. PMID- 15894420 TI - C-terminus of a long alpha-neurotoxin is highly mobile when bound to the nicotinic acetylcholine receptor: a time-resolved fluorescence anisotropy approach. AB - To better understand how alpha-neurotoxins interact with the acetylcholine receptor, four fluorescein isothiocyanate derivatives of the siamemsis alpha cobratoxin were prepared (conjugated to the epsilon-amino group in Lys(23), Lys(35), Lys(49), or Lys(69)) and the time-resolved fluorescence anisotropy of each conjugate was measured free in solution and bound to the Torpedo acetylcholine receptor. All the conjugated reporter groups displayed a high and comparable level of mobility free in solution. When receptor bound, on the other hand, significant differences in the conformational dynamics of the reporter groups were observed with the C-terminal Lys(69) derivative displaying by far the greatest mobility strongly suggesting that the C-terminal domain of the bound neurotoxin is highly mobile and does not participate in the toxin-nAChR binding surface. Additionally, this study demonstrates the utility of time-resolved fluorescence anisotropy to characterize the interaction of heteroproteins. PMID- 15894421 TI - Induction of pulmonary neoplasia in the smoke-exposed ferret by 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK): a model for human lung cancer. AB - Research into dietary chemoprevention against lung carcinogenesis has been limited by the lack of appropriate animal models that closely mimic smoking related human lung cancer. Ferrets (Mustela putorius furo) have been used to study the biologic activities of carotenoids against smoke-induced lung lesions, but this model has yet to be thoroughly established and validated. To determine the appropriateness of the ferret as a model for human lung cancer, we have performed a 6-month in vivo study in ferrets exposed to both tobacco smoke and a carcinogen (4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone, NNK) found in cigarette smoke. Results showed that six out 12 ferrets exposed to both NNK injection and cigarette smoke developed grossly identifiable lung tumors whereas none of nine ferrets from the sham treatment group developed any lung lesions. The histopathological types of these tumors (squamous cell carcinoma, adenosquamous carcinoma and adenocarcinoma) in ferret lungs are very similar to those in humans. In addition, 10 out of 12 ferrets exposed to both NNK and cigarette smoke developed preneoplastic lesions (squamous metaplasia, dysplasia, and atypical adenomatous hyperplasia) with complex growth patterns whereas the sham group did not show any of these lesions. Furthermore, the expression of proliferating cellular nuclear antigen increased markedly in both gross tumors and preneoplastic lesions in the lungs. In summary, the development of both preneoplastic lesions and gross lung tumors in ferrets provides an excellent and unique model for studying lung cancer chemoprevention with agents such as carotenoids, and for studying the molecular mechanism of carcinogenesis in the earlier stages of smoke-related lung cancer. PMID- 15894422 TI - Alterations of liver function test indices of filling station workers with respect of genetic polymorphisms of GSTM1 and GSTT1. AB - To determine the health effects of gasoline exposure on liver function test indices of filling station workers the present study was done. This case-control study was conducted in Shiraz on 56 male gasoline workers and 56 age- and sex matched control subjects with no occupational exposure to gasoline. To elucidate the role of hepatic detoxifying enzymes, the genotypes of glutathione-S transferases (GST) M1 and T1 were determined. Data analysis was done by multiple linear regression analysis and non-parametric Kruskal-Wallis test. The present study showed that all measurements were in normal range, although sub-clinical changes were detected in some indices. In liver function tests, exposure was associated with lower serum albumin (t=-3.88, P<0.001) and total proteins (t= 3.016, P=0.003) but higher alanine aminotransferase (t=2.856, P=0.005) and aspartate aminotransferase (t=2.11, P=0.038) levels in workers comparing to controls. Other investigators reported that GSTs involved in detoxification of several toxins including some of the compounds present in gasoline. Therefore, the possible influence of GSTT1 and GSTM1 genetic polymorphisms on alteration of liver function tests indices was investigated. The present findings showed that the genotype combinations of GSTM1 and GSTT1 did not alter the effects of exposure to gasoline in workers except for serum albumin. Serum albumin significantly decreased in workers with both active GST enzymes who had more than 5 years of employments (P=0.01). It is suggested that GSTM1 and GSTT1 are not involved in detoxification of toxicants present in gasoline which are hazardous to liver. Overall, due to detection of sub-clinical changes in hepatic test in gasoline station workers, exposure limitation and administrating safety device are recommended. PMID- 15894423 TI - Removal of lead (II) ions from synthetic and real effluents using immobilized Pinus sylvestris sawdust: adsorption on a fixed-bed column. AB - The purpose of this work was to evaluate the potential of Pinus sylvestris sawdust, in a continuous flow removal of lead (II) ions from synthetic and industrial aqueous effluents. The kinetic parameters obtained in a batch process were used to scale-up the process on a mini-column and to choose the breakthrough model. The column experimental data concerning the volumes treated were correlated using the bed depth service time model. These experimental data closely fitted the bed depth service time model at 10% of the breakthrough curve. The results from the bed depth service time model on the mini-column were then used to design a pilot plant adsorption unit. The performance of the pilot plant column accurately agreed with that obtained from the mini-column. The experiments carried out in a dynamic reactor allowed to bring out the influence of various parameters on the efficiency of the P. sylvestris sawdust. In addition, the process was checked for the treatment of industrial aqueous effluents on a pilot plant scale and the results were in accordance with those obtained from synthetic effluents. PMID- 15894424 TI - Human plasma semicarbazide sensitive amine oxidase (SSAO), beta-amyloid protein and aging. AB - Semicarbazide sensitive amine oxidase (SSAO) metabolizes oxidative deamination of primary aromatic and aliphatic amines. The final products of its catalysis, ammonia, hydrogen peroxide (H2O2) and the corresponding aldehyde, may contribute to diseases involving vascular degeneration. SSAO is selectively expressed in blood vessels in the brain, but is also present in blood plasma. We have previously reported that membrane-bound SSAO is overexpressed in the cerebrovascular tissue of Alzheimer's disease (AD) patients. The aim of the present work is to study whether the circulating SSAO is also altered in this neurodegenerative disease. SSAO activity was determined in plasma of control cases (n = 23) and patients suffering sporadic Alzheimer dementia, distributed according to the Global Deterioration Scale (GDS): mild (n = 33), moderate (n = 14), moderate-severe (n = 15) and severe dementia (n = 19). Results show a clear increase of plasma SSAO activity (p < 0.001) in moderate-severe and severe AD patients, with patient age being an independent correlative factor. However, plasma SSAO activity was not altered in AD patients with mild or moderate dementia compared to controls. beta-Amyloid (Abeta) (40-42) immunoreactivity in plasma samples was also determined, and no correlation was observed between Abeta 40-42 levels and the severity of the dementia or the plasma SSAO activity. Our results suggest that an increase in circulating SSAO activity could contribute to oxidative stress and vascular damage in advanced Alzheimer's disease. PMID- 15894425 TI - The flavanoide caffeic acid phenethyl ester blocks 6-hydroxydopamine-induced neurotoxicity. AB - Parkinson's disease (PD) is a neurodegenerative disorder characterized by progressive loss of dopaminergic (DA) neurons of the substantia nigra pars compacta. 6-Hydroxydopamine (6-OHDA) is specific to dopaminergic neurons in intrastriatal rodent models. It induces neuronal death either via uncoupling mitochondrial oxidative phosphorylation resulting in energy deprivation or alternatively, is associated with its ability to produce hydrogen peroxide, hydroxyl and superoxide radicals. Caffeic acid phenethyl ester (CAPE), an antioxidant flavanoid, has antiviral, anti-inflammatory, antioxidant, and immunomodulatory properties. Recent studies have shown that CAPE has also a neuroprotective effects in ischemia and low potassium-induced neuronal apoptotic models. In cerebellar granule neurons CAPE significantly blocks 6-OHDA mediated cell death (70 microM) in a dose-dependent manner. Furthermore, CAPE was able to modulate the Ca(2+)-induced release of cyctochrome c in isolated liver mitochondria. Caspase-3 activation following 6-OHDA treatment was markedly inhibited in the presence of CAPE. Although the molecular mechanisms associated with CAPE's neuroprotective effects remain to be elucidated in more detail, our results clearly demonstrate a considerable neuroprotective effect of CAPE. Since a mitochondrial insult is a major cause for the degeneration of nigral neurons in PD, we hypothesize that propolis derivatives, in particular CAPE, may have a neuroprotective effect on those cells and may be a promising drug candidate to be taken into in vivo models of PD. PMID- 15894426 TI - Event-related potentials suggest early interaction between syntax and semantics during on-line sentence comprehension. AB - Event-related potentials (ERPs) were used to investigate interaction between syntactic parsing and semantic integration processes during a visual sentence comprehension task. The linguistic stimuli were Finnish five-word sentences containing morphosyntactic and/or semantic violations. Single morphosyntactic violations evoked left anterior negativity (LAN) and P600 components. Single semantic violations elicited a robust N400 effect over the left hemisphere. A later and weaker N400-like response was also observed in the right hemisphere, left-right hemispheric latency difference being 40 ms. Combined morphosyntactic and semantic violations elicited a P600 component and a negative ERP component within the latency range of the LAN and N400 components. Further analysis of these ERP effects provided evidence for early processual interaction between syntax and semantics during on-line sentence comprehension. The hemispheric distribution of the LAN and N400 components was taken to suggest lateralization of initial morphosyntactic parsing and semantic integration processes to the left hemisphere. In contrast, the later syntax-related P600 component was observed as being more pronounced over the posterior areas of the right hemisphere. PMID- 15894427 TI - State of the Heartworm. Proceedings of the 11th Triennial Symposium of the American Heartworm Society, Philadelphia, Pennsylvania, USA, July 23-24, 2004. PMID- 15894428 TI - Engineering dihydropteroate synthase (DHPS) for efficient expression on M13 phage. AB - Phage display is a commonly used selection technique in protein engineering, but not all proteins can be expressed on phage. Here, we describe the expression of a cytoplasmic homodimeric enzyme dihydropteroate synthetase (DHPS) on M13 phage, established by protein engineering of DHPS. The strategy included replacement of cysteine residues and screening for periplasmic expression followed by random mutagenesis and phage display selection with a conformation-specific anti-DHPS antibody. Cysteine replacement alone resulted in a 12-fold improvement in phage display of DHPS, but after random mutagenesis and three rounds of phage display selection, phage display efficiency of the library had improved 280-fold. Most of the selected clones had a common Asp96Asn mutation that was largely responsible for the efficient phage display of DHPS. Asp96Asn affected synergistically with the cysteine replacing mutations that were needed to remove the denaturing effect of potential wrong disulfide bridging in phage display. Asp96Asn alone resulted in a 1.8-fold improvement in phage display efficiency, but in combination with the cysteine replacing mutations, a total of 130-fold improvement in phage display efficiency of DHPS was achieved. PMID- 15894429 TI - Fludarabine increases complete response but not survival compared with conventional alkylator-based regimens for previously untreated chronic lymphocytic leukaemia. PMID- 15894430 TI - The inevitable rise of mediocrity in academic medicine. AB - The present analysis addresses the question of how it has been possible for academic medicine to grow and lose its creative productivity at the same time. A mathematical model is developed to simulate the mechanisms that govern growth of medical systems over time. Time-dependent growth of system size increases the occurrence of statistical deviations of all system parameters. Deviations are correlated with costs and creative output. As deviation-induced costs start to strain the system's tolerance, means become implemented to restrict deviation, which ultimately also reduces its creative output. To maintain growth combined with high levels of creative output, an academic medical system would need to continuously branch off and nurture smaller subsystems, which pursue their own set of goals relatively independently of the overall academic structure. PMID- 15894431 TI - Ilex paraguariensis extracts inhibit AGE formation more efficiently than green tea. AB - Glycation, the nonenzymatic adduct formation between sugar dicarbonyls and proteins, is one key molecular basis of diabetic complications due to hyperglycemia. Given the link between glycation and oxidation, we hypothesized that herbal extracts with a high concentration of antioxidant phenolics might possess significant in vitro antiglycation activities as well. The aim of the present study was to address the hypothesis that polyphenol-rich Ilex paraguariensis (IP) extracts are capable of inhibiting advanced glycation end products (AGEs) formation and to compare the potency of these extracts with green tea and with the standard antiglycation agent aminoguanidine. When we studied the effects of IP extract on AGE fluorescence generated on bovine serum albumin (BSA) by glycation with methylglyoxal, a dose-dependent effect that reaches 40% at 20 mul/ml of extract was demonstrated. Green tea did not display any significant effect. IP polyphenols are about 2- to 2.5-fold higher in our preparations compared with green tea. The effect of IP, therefore, may be due not only to the higher concentrations but to the different composition in phenolics of the two botanical preparations as well. To better discriminate between an antioxidant or a carbonyl quenching mechanism of action, we explored tryptophan fluorescence and cross-linking by sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) electrophoresis. The conformational changes induced by glycation and substitution of positive charges in arginine and/or lysine produce a decrease in tryptophan fluorescence. We show that incubation of BSA with methylglyoxal produces dramatic changes in tryptophan fluorescence that are prevented by aminoguanidine. This also prevents the downstream effect of AGE formation. Neither green tea nor IP extracts displayed any significant effect which rules out any significant participation as inhibitors in the first phase of the glycation cascade. The results from the SDS-PAGE serve to confirm the above-mentioned data. The effect is therefore due mainly to an inhibition of the second phase of the glycation reactions, namely the free-radical mediated conversion of the Amadori products to AGE. Taken together our results demonstrate a significant, dose-dependent effect of water extracts of I. paraguensis on AGE adducts formation on a protein model in vitro, whereas green tea displays no significant effect. The inhibition of AGE formation was comparable to that obtained by using millimolar concentrations of the standard antiglycation agent aminoguanidine. PMID- 15894432 TI - What were they thinking? Adolescents' interpretations of DSM-IV alcohol dependence symptom queries and implications for diagnostic validity. AB - OBJECTIVE: DSM-IV alcohol dependence criteria of tolerance to alcohol and drinking more or longer than intended have relatively high prevalence among youth, and may be vulnerable to false positive symptom assignments that degrade diagnostic validity. We conducted a methodological study of DSM-IV symptom queries used to assess alcohol tolerance and impaired control over drinking to determine potential sources of measurement error. METHOD: Adolescents recruited from addictions treatment participated in either a focus group (n = 9) or an individual interview (n = 41) to provide data on their interpretation of selected items contained in a semi-structured diagnostic interview. RESULTS: When alcohol tolerance is operationally defined as a change in quantity to obtain the same effect, large individual differences in the change in quantity that represents a high level of tolerance limit the utility of this operational definition as an indicator of dependence. The symptom "drinking more or longer than intended", includes the embedded assumption that a limit on use had been set. Teens, however, typically intended to become intoxicated, rather than to keep to a limit. CONCLUSIONS: Adolescents' understanding of symptom queries suggests how validity of DSM-IV alcohol symptoms and diagnoses can be improved through greater attention to developmental considerations affecting assessment. PMID- 15894433 TI - Rating the severity and character of transient cocaine-induced delusions and hallucinations with a new instrument, the Scale for Assessment of Positive Symptoms for Cocaine-Induced Psychosis (SAPS-CIP). AB - BACKGROUND: Cocaine can induce transient psychotic symptoms. We examined the phenomenology of such cocaine-induced psychosis (CIP) using a modified version of the Scale for Assessment of Positive Symptoms (SAPS), a well-validated instrument for the assessment of schizophrenic psychosis. METHODS: We developed a new instrument, the Scale for Assessment of Positive Symptoms for Cocaine-Induced Psychosis (SAPS-CIP), based on the well-validated SAPS. We interviewed 243 unrelated cocaine-dependent adults using both the SAPS-CIP and an instrument for the identification of cocaine-induced paranoia, the Cocaine Experience Questionnaire (CEQ). RESULTS: One hundred and eighty-one (75%) of the subjects endorsed CIP using the CEQ. With the SAPS-CIP, hallucination (HAL) and delusion (DEL) scores correlated strongly, and the DEL domain showed excellent concurrent validity with the CEQ. We observed significant positive correlations, respectively, between severity of HAL and DEL, and lifetime number of episodes of cocaine use, and negative correlations with age at onset of cocaine use. CONCLUSIONS: The results suggest that CIP consists of transient delusional and hallucinatory symptoms, which tend to occur together and co-vary in severity. It appears that rating cocaine-induced paranoia alone (e.g., with the CEQ) can identify most subjects experiencing CIP. However, the SAPS-CIP is useful for quantifying the severity of CIP according to operational criteria. Our data provide additional evidence that CIP is a sensitizing response. PMID- 15894434 TI - Contrasting predictors of readiness for substance abuse treatment in adults and adolescents: a latent variable analysis of DATOS and DATOS-A participants. AB - Although readiness for substance abuse treatment is a potent predictor of short term clinical response, predictors of readiness are still poorly understood. Predictive models are customarily not predicated on dependence or abuse, but usually focus instead on intrapsychic motivational change or external contextual incentives. Based on a secondary analysis of the drug abuse treatment outcome studies (DATOS) data set for adults and a similar data set for adolescents (DATOS A), latent variables were constructed to represent readiness for treatment, various psychosocial and behavioral features of substance use, substance-related problems and specific substance use frequency. Identical measures were used for each group. The two groups were then directly compared using a series of constrained covariance structure models. A path model using all hypothesized latent variables, treatment modality and demographics predicted Readiness for treatment in each separate group. Substance problems, a latent variable reflecting cross-substance symptoms of dependence and abuse, was the strongest predictor of Readiness for treatment in both age groups. Health problems and deviant family/peers were also strong predictors of readiness in both age groups. Demographic variables and specific substances used predicted readiness directly and indirectly in an age-specific manner. This study links readiness for substance abuse treatment to the core disorders targeted by that treatment in adults and adolescents for the first time. PMID- 15894435 TI - Mitochondrial DNA impacts the morphology of mitochondrial compartments. AB - Mitochondrial compartments of the yeast Saccharomyces cerevisiae experience continual morphological alterations. Mitochondrial compartments of wild-type yeast, when observed using fluorescent markers, are usually found to be a network of extended tubular structures. However, a quantitative analysis of mitochondrial structures in a genetically homogenous population of wild-type yeast revealed that although the majority of individual yeast cells contained the expected extended network of mitochondrial tubules, a significant number of cells were found to exclusively contain condensed globular mitochondrial compartments or a mixture of extended and globular mitochondrial compartments. Additionally, this distribution of mitochondrial morphologies was found to be dependent upon the presence of mitochondrial DNA. Cells containing intact wild-type genomes or a deletion mutation of the COX2 gene gave rise to populations of yeast in which at least 80% of the cells contained only extended tubular networks of mitochondria. In isogenic yeast strains lacking mitochondrial DNA or containing a mitochondrial genome composed of reiterated COX2 sequences, only 30 to 40% of the cells in the population had exclusively extended mitochondrial networks, and the remaining cells in the population were composed of cells exhibiting either exclusively condensed or both condensed and extended mitochondrial profiles. We conclude that either a specific sequence element or a mitochondrially encoded gene product is required for promoting a pervasive distribution of extended tubular mitochondrial compartments. PMID- 15894436 TI - Acid stress adaptation protects Saccharomyces cerevisiae from acetic acid-induced programmed cell death. AB - In this work evidence is presented that acid stress adaptation protects Saccharomyces cerevisiae from acetic acid-mediated programmed cell death. Exponential-phase yeast cells, non-adapted or adapted to acid stress by 30 min incubation in rich medium set at pH 3.0 with HCl, have been exposed to increasing concentrations of acetic acid and time course changes of cell viability have been assessed. Adapted cells, in contrast to non-adapted cells, when exposed to 80 mM acetic acid for 200 min did not display loss of cell viability associated to morphological alterations typical of apoptosis. Thus, 80 mM acetic acid death inducing conditions were selected to further characterize the early molecular events leading to such active cell death process. Catalase was specifically activated during acid stress adaptation and protection against acetic acid induced death was associated with maintenance of its activity during treatment with 80 mM acetic acid. On the other hand, intracellular superoxide dismutase activity was found present at comparable levels both in adapted and in dying yeast cells, excepting in non-adapted cells which displayed a maximum activity value after 15 min acetic acid exposure, corresponding to more than 80% cell viability. This study gives first experimental evidence that H2O2, rather than superoxide, detoxification may have a major role in preventing yeast cell death in response to acetic acid. The results, as a whole, suggest that commitment of S. cerevisiae to a programmed cell death process in response to acetic acid is mediated through a ROS-dependent apoptotic pathway. PMID- 15894437 TI - The silk moth Bombyx mori U1 and U2 snRNA variants are differentially expressed. AB - Five U1 and eight U2 isoforms of the silk moth Bombyx mori exhibiting internal nucleotide differences have been previously identified and characterized in various tissues and developmental stages. In this investigation, it is demonstrated that the levels of some snRNA variants differ in egg and silk gland tissue and change during development. Qualitative and quantitative differences in the U1 and U2 variant populations were observed at three developmental points (early, middle and late) of the silk gland (SG) during the fifth instar larval stage of the silk moth. Statistical analyses of the various isoform populations across the fifth instar larval and egg stages show significant differences for some of the U1 and U2 variants. The representation of variant sequences in expressed U1 and U2 sequences (RT-PCR libraries) and in a whole-genome shotgun (WGS) assembly database was confirmed. In addition, conserved elements in the promoter 5'-flanking region of the U1 and U2 variants were identified in the WGS. PMID- 15894438 TI - Effects of low-dose perinatal cadmium exposure on tissue zinc and copper concentrations in neonatal mice and on the reproductive development of female offspring. AB - It has been suggested that the toxic effects of cadmium (Cd) are the result of interactions with essential metals, such as zinc (Zn) and copper (Cu). Previous studies have shown altered Zn and/or Cu levels in the tissues of rodents that drank water supplemented with >50 ppm Cd. To evaluate the effects of lower level Cd exposure on maternal and neonatal Zn and Cu levels and on the reproductive organs of female offspring, mice were exposed to 0, 1 and 10 ppm Cd in the drinking water from conception to 10 days after birth. The Cd concentrations in the brains of the offspring were higher in the exposed group than in the control group at birth. In the kidneys and livers, the Cd concentrations were higher in the Cd-exposed group 10 days after birth. At birth, increased Zn concentrations were observed in the kidneys and livers of the Cd-exposed offspring, although the Cd concentrations in these tissues did not differ between the exposed and non exposed groups. The hepatic Cu concentrations of the exposed mice tended to be lower than those of the control mice at birth and were significantly lower 10 days after birth. In addition, Cd exposure tended to delay the timing of vaginal opening and perturbed the estrous cycles of the female offspring. These findings suggest that perinatal Cd exposure, even at low levels, affects the Zn and Cu concentrations of neonates and the reproductive functions of female offspring. PMID- 15894439 TI - Effects of teat number on litter size in gilts. AB - This study was carried out to investigate the effects of teat number and interval at first estrus and mating on litter size in Duroc, Landrace and Yorkshire gilts. Gilt body weight at first estrus was from 101.5 kg to 115.3 kg and gilts normally attained puberty at 170.5-181.5 days of age. Breed differences among Duroc, Landrace and Yorkshire in body weight and age at first estrus and mating were found. Total teat number of Duroc, Landrace and Yorkshire were 12.5, 14.9 and 13.7, respectively. Teat interval from pectoral to inguinal region and from left to right at first estrus and mating did not show any differences among the breeds. In conclusion, 14 or more teat number compared to 11-13 teat number in gilts increased litter size at birth and at 21 day weaning. PMID- 15894440 TI - Sexual function in menopausal women in Kelantan, Malaysia. AB - OBJECTIVES: The aim of the study was to document sexual function in Kelantanese postmenopausal women. METHOD: A semi-structured questionnaire in Malay language was administered to 326 women (mean age of 57.1+/-6.58 (S.D.) years) residing in Kelantan. The subjects comprised of naturally menopaused, healthy women. RESULTS: Of the total respondents, 70% (n=227) were with a spouse at the time of the study. Of these, more than two-thirds reported a decrease in sexual activity following menopause. Varying degree of dyspareunia was reported by 44% of the women. A small fraction (8.8%) reported inability of the vagina to stretch sufficiently to enable the complete penetration of an erect penis. Of the total married respondents, vaginal secretion during sexual intercourse was decreased in 52.4%, did not change in 31% but increased in 1.3% of the women following menopause. Sexual desire was reportedly decreased or absent in two-thirds of the total respondents (n=326). CONCLUSION: It appears that sexual function significantly decreases during menopause. This may be due to dyspareunia, poor lubrication, loss of sexual desire, and the spouse's health status and ageing itself. Although declining sexual function was recognised by nearly two-thirds of the women, more than half did not take any action to improve their sexual function. Of those who did, they used hormonal therapy, traditional, alternative medicine or practiced healthy lifestyle or a varied combination of above self help actions. PMID- 15894441 TI - Effects of tibolone on selectins in postmenopausal women. AB - OBJECTIVE: The first step in atherosclerosis is characterized by the adherence of lymphocytes and monocytes to cell adhesion molecules expressed by endothelial cells. The precise mechanism by which steroid hormones may be exerting a protective action against atherogenesis remains unclear. Therefore, we wanted to investigate the effect of tibolone on the circulating levels of various selectins in postmenopausal women. METHODS: Thirty healthy postmenopausal women were enrolled in a prospective, randomized, double blind, placebo-controlled outpatient trial. RESULTS: Patients treated with tibolone revealed a significant decrease for the variables sE-selectin, sL-selectin, and sPECAM-1 after 8 weeks of treatment. CONCLUSIONS: By reducing leukocyte adhesion molecule expression on human endothelial cells, tibolone may have the intrinsic potential to exert additional, lipid-independent, cardiovascular protective effects that may explain the clinical benefits of cardiovascular diseases in postmenopausal women. PMID- 15894442 TI - The expression of the hormone receptors in the endometrium and endometrial polyps in postmenopausal women and its relationship to body mass index. AB - OBJECTIVE: To evaluate and compare the relationship of body mass index (BMI) and the immunoexpression of estrogen (ER) and progesterone receptors (PR) in endometrial polyps (EP) and endometrium, in gland and stromal cells of postmenopausal women. METHODS: Thirty-five postmenopausal women with benign endometrial polyps, who had not been taking medication with hormonal effects for at least 6 months, were submitted to operative hysteroscopy. The presence of ER and PR were evaluated by immunohistochemical method using a semiquantitative analysis. RESULTS: BMI was significantly higher among patients with lower expression of ER in the glands of endometrium (p=0.02). EP and adjacent endometrium showed significantly higher proportion of positive cells in the glands than in the stroma, for both ER (p=0.0015 and 0.0018, respectively) and PR (p=0.0176 and p<0.0001, respectively). Glands and stroma cells showed significantly higher proportion of positive cells in polyps than in the endometrium, for ER (p<0.0001 and p=0.0034, respectively). CONCLUSIONS: The higher proportion of positive gland cells for ER in EP as compared to endometrium supports an implication of these receptors in the pathogenesis of polyps. Association of higher BMI with lower expression of ER in endometrial glands, but not in EP, may indicate that factors influencing ER expression do not affect EP, supporting an autonomous function of polyps. PMID- 15894443 TI - In vitro evaluation of coating polymers for enteric coating and human ileal targeting. AB - Recombinant interleukin-10 producing Lactococcus lactis is an alternative therapy for Crohn's disease. For in vivo interleukin-10 production, thymidine, the essential feed component of these recombinant bacteria should be coadministered. Different coating polymers were evaluated in vitro for enteric properties and targeting suitability to the ileum, the major site of inflammation in Crohn's disease. To guarantee ileal delivery, the polymer must dissolve from pH 6.8 and allow complete release within 40 min. Aqoat AS-HF coated pellets (15%) showed poor enteric properties and thymidine was released below pH 6.8. Eudragit FS30D coated pellets (15%) showed good enteric properties, but no thymidine was released within 40 min at pH 6.8. Eudragit S coated pellets (15%) showed good enteric properties after curing at elevated temperature while no thymidine was released within 40 min at pH 6.8. In another approach to pass the proximal small intestine intact, pellets were coated with 30% Eudragit L30D-55. At pH 6.0, they showed a lag-phase of 20 min. No influence of layer thickness was seen above pH 6.5. Alternatively, pellets were coated with a mixture of Eudragit FS30D/L30D-55 but they showed poor enteric properties and thymidine was released below pH 6.8. In conclusion, none of the tested polymers/mixtures ensured enteric properties and ileal targeting. PMID- 15894444 TI - Anti-diabetic effect on alloxanized and normoglycemic rats and some pharmacological evaluations of Tournefortia hartwegiana. AB - The purpose of the present investigation was to assess the pharmacological properties of Tournefortia hartwegiana Steud (Boraginaceae), used in traditional medicine for the treatment of diabetes, diarrhea and kidney pain in Morelos, Mexico. Administration of methanol extract from aerial parts of Tournefortia hartwegiana (310 mg/kg body weight/day) for 10 days, to normoglycemic and alloxan induced diabetic rats, significantly lowered their blood glucose levels (37 and 36%, respectively, p<0.05). The anti-diabetic and hypoglycemic activities due to the MeOH extract were similar to those produced by metformin at 120 mg/kg (positive control, p<0.05). In contrast, the hexane, dichloromethane and MeOH extracts from the same species showed no significant spasmolytic effect and did not have activity in antibacterial and Artemia salina toxicity bioassays. PMID- 15894445 TI - Genetic profile of nine STR loci among Goud and Padmashali populations of Andhra Pradesh, India. AB - We have analysed nine autosomal STR loci (D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820) in Goud and Padmashali populations belonging to Chittoor and Cuddapah districts of Andhra Pradesh, India, respectively. This data was compared with the available data on Indian populations. Heterozygosity (H), power of discrimination (PD), probability exclusion (PE), typical paternity index (TPI), polymorphism information content (PIC), exact test with Bonferroni corrections and Arlequin analysis were carried out. Average heterozygosity observed in Goud and Padmashali populations were 0.82 and 0.81, respectively. Similarly, PD, PE, TPI and PIC have been found to be almost same between these two populations. Overall data analysis suggests the existence of sub-structuring in Indian populations. PMID- 15894446 TI - Percutaneous gastrostomy in patients with complete obstruction of the upper digestive tract. AB - Gastrostomy for nutritional support can be performed radiologically when endoscopic technique is not possible. In patients with complete obstruction of the upper digestive tract, the radiological technique in its conventional method may not be possible, as insertion of naso-gastric tube for gastric insufflation is not always successful. We reviewed our experience of gastrostomy insertion in nine such patients after failure of the conventional method. In seven of the nine patients, initial gastric puncture was achieved with a 22G needle under direct ultrasound visualisation. In the remaining two patients, initial puncture was made into locules of gas in the stomach with fluoroscopy. The stomach was then distended with air and a gastrostomy tube inserted by conventional technique. We conclude that percutaneous gastrostomy can be undertaken safely even in the presence of complete obstruction of the upper digestive tract and recommend this technique in this selective group of patients. PMID- 15894447 TI - Development of a rapid HPLC method for determination of famotidine in human plasma using a monolithic column. AB - A rapid and sensitive HPLC method using a monolithic column has been developed for quantification of famotidine in plasma. The assay enables the measurement of famotidine for therapeutic drug monitoring with a minimum detectable limit of 5 ngml(-1). The method involves simple, one-step extraction procedure and analytical recovery was complete. The separation was carried out in reversed phase conditions using a Chromolith Performance (RP-18e, 100 mm x 4.6 mm) column with an isocratic mobile phase consisting of 0.03 M disodium hydrogen phosphate buffer-acetonitrile (93:7, v/v) adjusted to pH 6.5. The wavelength was set at 267 nm. The calibration curve was linear over the concentration range 20-400 ngml( 1). The coefficients of variation for inter-day and intra-day assay were found to be less than 8%. PMID- 15894448 TI - Simultaneous determination of andrographolide and dehydroandrographolide in Andrographis paniculata and Chinese medicinal preparations by microemulsion electrokinetic chromatography. AB - The present paper describes the development of a microemulsion electrokinetic chromatographic (MEEKC) method for simultaneous determination of andrographolide and dehydroandrographolide in traditional Chinese medicines and Chinese medicinal preparations. The MEEKC method involved the use of sodium dodecyl sulfate (SDS) as surfactant, heptane as organic solvent and butan-1-ol as co-solvent. The effect of temperature and pH of running buffers on separation were examined. The optimized conditions (heptane 0.81% (w/w), SDS 3.31% (w/w), butan-1-ol 6.61% (w/w) and 10mM sodium tetraborate buffer, pH 9.2) allowed a useful and good reproducible separation of the studied analytes to be achieved. PMID- 15894449 TI - Stability of baicalin in biological fluids in vitro. AB - The stability of baicalin in buffered aqueous solutions at different pHs and in biological fluids, including plasma, urine and tissue homogenates, were investigated in vitro. Structures of the degradation products of baicalin were elucidated by liquid chromatography-electrospray ion trap mass spectrometry and the degradation mechanism was proposed with the aid of electron paramagnetic resonance spectrometry. The results showed that the degradation of baicalin was pH- and temperature-dependent. The oxidation-reduction reaction intermediated by phenoxyl radicals is the major degradation process for baicalin in plasma and urine in vitro, whereas baicalin mainly undergoes hydrolysis and phase II metabolic pathways when spiked in tissue homogenates. It is found that acidification can stabilize baicalin in urine and plasma (the final pH adjusted to 3.0-4.0), and baicalin is relatively stable in tissues homogenized with methanol. PMID- 15894450 TI - Separation and simultaneous determination of rutin, puerarin, daidzein, esculin and esculetin in medicinal preparations by non-aqueous capillary. AB - A simple method for the simultaneous determination of five bioactive components (rutin, puerarin, daidzein esculin and esculetin) in traditional medicinal preparations by non-aqueous capillary electrophoresis with UV detection has been developed for the first time. A running buffer composed of 15% acetonitrile, 2.5% acetic acid and 90 mM sodium cholate in methanol was found to be the most suitable for this separation. The limits of detection for five analytes were over the range of 0.050-1.216 microg ml(-1). The relative standard deviations (R.S.Ds.) of the migration times and the peak areas of the analytes were in the range of 1.3-2.9% and 2.2-2.7% (intraday), 1.7-1.9% and 2.8-3.6% (interday), respectively. In the tested concentration range, linear relationships (correlation coefficients: 0.9974 for rutin, 0.9976 for puerarin, 0.9981 for daidzein, 0.9972 for esculin and 0.9929 for esculetin) between peak areas and concentrations of the analytes were obtained. This method has been successfully applied to simultaneous determination of the five bioactive components with recoveries over the range of 89.4-107.4%. PMID- 15894451 TI - Investigation of the retention/pH profile of zwitterionic fluoroquinolones in reversed-phase and ion-interaction high performance liquid chromatography. AB - The retention/pH profiles of three fluoroquinolones, ofloxacin, norfloxacin and ciprofloxacin, was investigated by means of reversed-phase high performance liquid chromatography (RP-HPLC) and reversed-phase ion-interaction chromatography (RP-IIC), using an octadecylsilane stationary phase and acetonitrile as organic modifier. Sodium hexanesulphonate and tetrabutylammonium hydroxide were used as sources of counter ions in ion-interaction chromatography. The retention/pH profiles under in RP-HPLC were compared to the corresponding lipophilicity/pH profiles. Despite the rather hydrophilic nature of the three fluoroquinolones positive retention factors were obtained while there was a shift of the retention maximum towards more acidic pH values. This behavior was attributed mainly to non hydrophobic silanophilic interactions with the silanized silica gel material of the stationary phase. In ion-interaction chromatography the effect of counter ions over a broad pH range was found to be ruled rather by the ion pair formation in the mobile phase which led to a drastic decrease in retention as a consequence of the disruption of the zwitterionic structure and thereupon the deliberation of a net charge in the molecules. At pH values at which zwitterionic structure was not favored both the ion-exchange and ion pair formation mechanisms were assumed to contribute to the retention. PMID- 15894452 TI - Prediction of protein retention in hydrophobic interaction chromatography. AB - Hydrophobic interaction chromatography (HIC) is a powerful technique for protein separation. This review examines methodologies for predicting protein retention time in HIC involving elution with salt gradients. The methodologies discussed consider three-dimensional structure data of the protein and its surface hydrophobicity. Despite their limitations, the methods discussed are useful in designing purification processes for proteins and easing the tedious experimental work that is currently required for developing purification protocols. PMID- 15894454 TI - The prognostic role of the sentinel lymph node in clinically node-negative patients with cutaneous melanoma: experience of the Genoa group. AB - AIM: To define the benefit of intraoperative frozen section examination of the sentinel lymph node (sN), and to assess its prognostic value in clinically node negative melanoma patients. MATERIALS AND METHODS: Between July 1993 and December 2001, 214 patients with Stage I-II cutaneous melanoma underwent sN biopsy; complete follow-up data are available in 169 of 175 patients who underwent preoperative lymphoscintigraphy, lymphatic mapping with Patent Blue-V and radio guided surgery (RGS). RESULTS: In an initial subset, the sN was identified in 35 out of 39 patients; in the principal group of 169 patients, the sN was detected in all patients. The benefit of frozen section examination, that is the proportion of all patients having intraoperative histologic examination who tested positive, was 17.2% (29/169); notably, in patients with pT(1-2) vs pT(3-4) melanoma the corresponding values were 2.3 and 33.3%, respectively, (P=0.000). Cox regression analysis for overall survival indicated that sN-positive patients had a two-fold increased risk of death; the most significant predictors of relapse-free survival were sN status (P=0.004), age (P=0.015), and T stage grouping (P=0.033). CONCLUSIONS: The sN is a reliable predictor of regional lymph node status in patients with cutaneous melanoma. Frozen section examination can be useful in avoiding a 'two-stage' operative procedure in patients with tumour positive sN, but its greatest benefit seems to be restricted to patients with pT(3)-pT(4) primary melanoma. PMID- 15894455 TI - An examination of four models predicting fatigue in multiple sclerosis. AB - Fatigue is a common symptom of multiple sclerosis (MS) that is purported to cause significant distress and have detrimental effects on daily functioning, social and occupational obligations, and overall well-being. The prevalence of fatigue in MS is high, with 53-87% of patients reporting significant problems with fatigue across different studies reported in the literature. The cause of fatigue in MS is still poorly understood. Some researchers have suggested that fatigue is a direct consequence of the MS disease process, but several studies have failed to find a relationship between disease severity and MS fatigue. A number of investigations have reported that depression and sleep are significantly related to fatigue in MS, as well as to one another. The purpose of the present investigation was to examine the relationships among disease severity, depression, and sleep disturbance in MS, and their possible role in predicting fatigue. Four models were proposed to explore these relationships. The best fitting model showed that all three were significant independent contributors to fatigue in MS, accounting for 43% of the variance, with sleep disturbance reigning as the largest contributor. Furthermore, although disease severity predicted fatigue in our sample, both depression and sleep disturbance emerged as stronger predictors. These findings suggest that, beyond core physical/neurological MS symptomatology, there are other factors that contribute to fatigue in MS, namely, depression and sleep disturbance. PMID- 15894456 TI - Performance of forensic and non-forensic adult psychiatric inpatients on the Test of Memory Malingering. AB - This study compared performance on the Test of Memory Malingering (TOMM [Tombaugh, T. N. (1996). Test of Memory Malingering (TOMM). New York: Multi Health Systems]) between a Forensic Psychiatric group and a Non-forensic Psychiatric group of 20 men each. It was hypothesized that the Forensic group would perform less well on the TOMM due to greater secondary gain for that population. The Forensic group (age, M=32.65 years; 16/20 were minorities) was composed of inpatients from a forensic psychiatric facility who had been referred for pre-trial evaluations. The Psychiatric group (age, M=41.00 years; 15/20 were Caucasian) were chosen from an inpatient psychiatric facility and had no pending legal involvement. As hypothesized, the Psychiatric group performed significantly better than the Forensic group on all TOMM trials. A TOMM score of below 45 on Trial 2 or the Retention Trial is consistent with probable response bias. Only one member of the Psychiatric group (the same individual) met this criterion, whereas seven members of the Forensic group met this criterion. The TOMM identified patients with pending legal charges as more likely to exert less effort than those with no obvious secondary gain. PMID- 15894457 TI - Genomic annotation and expression analysis of the zebrafish Rho small GTPase family during development and bacterial infection. AB - The zebrafish genomic sequence database was analyzed for the presence of genes encoding members of the Rho small GTPases. The analysis shows the presence of 32 zebrafish Rho genes representing one or more homologs of the human RHOA, RND3, RHOF, RHOG, RHOH, RHOJ, RHOU, RHOV, CDC42, RAC1, RAC2, RAC3, RND1, RHOBTB1, RHOBTB2, RHOBTB3, and RHOT1 genes. By expression analysis using reverse transcriptase-PCR we show that at least 20 of the predicted zebrafish small GTPase genes are expressed in the adult stage. Interestingly, only 5 of these were found to be expressed at early embryonic stages, including rhoab, rhoad, cdc42a, cdc42c, and rac1a. We observed a strong upregulation of zebrafish rhogb expression after Mycobacterium marinum infection of adult fish. This complete annotation study provides a firm basis for the use of zebrafish as a model for analysis of Rho GTPase function in vertebrate development and the innate immune system. PMID- 15894458 TI - A temporal framework for understanding the effects of stressful life events on inflammation in patients with multiple sclerosis. AB - A growing literature reports that stressful life events are associated with exacerbation and the subsequent development of brain lesions in patients with multiple sclerosis (MS). The evolution an MS exacerbation occurs over a period of many months and involves many different biological processes that change over time. Likewise, the experience of stress also occurs over time, with an onset, a shift from acute to chronic in some cases, and resolution. Each of these phases is associated with unique biological features. Thus, the impact of stress on MS exacerbation may depend on the temporal trajectories of stress and MS exacerbation, and when the intersection between these two trajectories occurs. This paper presents a temporal model, along with three different temporal relationships and associated mechanisms by which stress may impact MS exacerbation. These include the onset of a stressor, which may be mediated by mast cell activation, the point that a stressor begins to become chronic, which may be mediated by glucocorticoid resistance in immune cells, and the resolution of the stressor, which may be mediated by a drop in cortisol. These three hypotheses are not necessarily mutually exclusive. Data on psychosocial mediators and moderators are also briefly reviewed and future research directions are discussed. PMID- 15894459 TI - Failed surgery for temporal lobe epilepsy: predictors of long-term seizure-free course. AB - OBJECTIVES: To identify prognostic factors which predict the outcome 2 years after TLE surgery in those patients who were not seizure-free at the 6-month postoperative examination. METHODS: We included 86 postoperative TLE patients who had undergone presurgical evaluation, including video-EEG and high-resolution MRI, and who had seizures between the second and sixth postoperative months. RESULTS: 32% of patients were seizure-free in the second postoperative year. We found that normal MRI findings and secondarily generalized seizures (SGTCS) preoperatively were associated with a non-seizure-free outcome, while rare postoperative seizures and ipsilateral temporal IED with seizure-free outcome. Newly administered levetiracetam showed a significant positive effect on the postoperative outcome independent of other prognostic factors. Five of seven patients who received levetiracetam became seizure-free (p = 0.006). CONCLUSION: One-third of patients who did not become seizure-free immediately after surgery, eventually achieved long-term seizure freedom. We suggest watching for long-term seizure freedom after failed epilepsy surgery especially in patients who had rare postoperative seizures, focal MRI abnormality, ipsilateral temporal spikes, or no SGTCS preoperatively. Levetiracetam may have a positive effect on postsurgical seizures. PMID- 15894460 TI - Patient characteristics and the likelihood of initiation on olanzapine or risperidone among patients with schizophrenia. AB - Although pharmacologic treatments are available for patients with schizophrenia, little is known about how prescription patterns of atypical antipsychotic agents are related to patient characteristics. In this study, we examined the association between patient characteristics and the likelihood of being initiated on olanzapine or risperidone, two of the most frequently prescribed atypical agents for schizophrenia. We selected patients who were diagnosed with schizophrenia or schizoaffective disorder based on > or = 1 inpatient or > or = 2 outpatient ICD-9-CM codes (> or = 7 days apart) between 7/1/98 and 6/30/99 from the Veterans Health Administration (VA). We classified patients into one of three types of initiation: (a) not on olanzapine or risperidone, (b) not on any atypical agents, or (c) not on any antipsychotic agents for 6 months, and then subsequently being prescribed the target drugs. Using logistic regression, we examined whether the odds ratio of being initiated on olanzapine versus risperidone are related to patient sociodemographic and clinical characteristics. Compared to risperidone initiators, olanzapine initiators used more drugs for psychiatric conditions (including antiparkinsonian agents, typical antipsychotics, and mood stabilizers) than risperidone initiators. On the other hand, risperidone initiators had more medical comorbidities and more non psychiatric hospitalizations. Olanzapine and risperidone appear to be prescribed to patients with different characteristics. Initiation of risperidone was more common among patients who presented with more medical comorbid conditions, whereas initiation of olanzapine was more common among patient who presented with more mental comorbid conditions. Future research needs to determine the reasons for those differences. PMID- 15894461 TI - Depressed mood and its functional correlates in institutionalized schizophrenia patients. AB - OBJECTIVE: To determine the frequency of depressed mood in institutionalized schizophrenia patients and its association with illness-related and functional variables. METHODS: Out of 657 institutionalized schizophrenia patients, patients with depressed mood were identified and compared to non-depressed patients, matching for potential confounders. RESULTS: Forty-eight (7.3%) patients had moderate to severe depressed mood. They were younger, more educated and had fewer years since their first hospitalization than non-depressed patients. After matching for these variables, depressed patients showed more positive symptoms and exhibited better social and cognitive functioning. When controlling for negative symptoms, the differences in social and cognitive functioning between the depressed and non-depressed patients disappeared, and depressed patients showed more positive symptoms and more impaired impulse control. CONCLUSIONS: Unlike the negative impact of depressed mood in other populations, this study shows that symptoms of depressed mood may identify a subgroup of institutionalized schizophrenia patients who show better functioning across a variety of indicators. Future studies should determine differential treatment responses and long-term outcomes of these patients. PMID- 15894462 TI - Xerosis in primary Sjogren syndrome: immunohistochemical and functional investigations. PMID- 15894463 TI - Hepatocyte growth factor in chronic leg ulcers--no biological activity--no improvement. PMID- 15894464 TI - Assessment of the efficacy of telithromycin simulating human exposures against S. pneumoniae with ribosomal mutations in a murine pneumonia model. AB - Telithromycin (TEL) is a ketolide antimicrobial agent with in vitro activity against Streptococcus pneumoniae (SPN), including macrolide resistant strains. The purpose of this study was to assess the efficacy of TEL against clinical SPN isolates with various genotypic mutations including the newly recognized ribosomal mutations. Pneumonia was induced in either immunocompetent and immunosuppressed mice. Six isolates were included in the study and all were resistant to azithromycin (AZI) by MIC testing. Three oral regimens of TEL were chosen to simulate the human pharmacokinetic (PK) exposures observed in young healthy, healthy elderly (> or =65 years), and infected subjects. An additional group was given AZI in human simulated doses. Bacterial density in lung was determined after each treatment. Telithromycin administered simulating infected patients showed greater efficacy (i.e., change in logCFU) than the azithromycin treated group for all isolates except P1660008. The immune system was responsible for increased efficacy (ranging from 45-146%) for all but one of the telithromycin treatment regimens. Unlike other isolates studied in this in vivo model, P1660008 displayed a highly variable response to therapy, such that the reductions in CFU were not consistent with the microbiological and PK profiles of either compound. For all other isolates, the activity of AZI was comparable with untreated controls. Human simulated exposures of TEL displayed 0.5-3.4 log kill in vivo despite the ribosomal mutations studied. These data support the in vivo efficacy of TEL against a variety of genotypic resistance profiles observed in pneumococci, however, additional studies are required to fully characterize the killing profile of the compound against these recently determined ribosomal mutations. PMID- 15894465 TI - Bactericidal activity of oral beta-lactam antibiotics in plasma and urine versus isogenic Escherichia coli strains producing broad- and extended-spectrum beta lactamases. AB - Bacteria harbouring extended-spectrum beta-lactamases (ESBLs), derived by mutation from TEM-1, TEM-2 or SHV-1 beta-lactamases, have been described world wide. The in vitro activities of these enzymes against beta-lactam antibiotics, including oral cephalosporins, are well recognised. The aim of this investigation was to assess the bactericidal activity of oral beta-lactam antibiotics available in Croatia (amoxicillin/clavulanate, cephalexin, cefuroxime, cefadroxil and ceftibuten), in biological fluids against isogenic Escherichia coli strains producing broad-spectrum (TEM-1, TEM-2 and SHV-1) and extended-spectrum beta lactamases (SHV-2, SHV-3, SHV-4, SHV-5, SHV-12). Bactericidal activity of oral beta-lactams in plasma and urine was tested in time-kill experiments and by determining bactericidal titres at different time intervals post-dose. The killing rate of antibiotics in urine was slower than in plasma, but faster than in Mueller-Hinton broth. High bactericidal titres in urine were only maintained throughout the whole dosing interval by ceftibuten against strains producing broad-, SHV-2 and SHV-3 beta-lactamases. The older generation cephalosporins can be considered for the therapy of urinary tract infections caused by E. coli harbouring TEM-1, TEM-2 and SHV-1 beta-lactamases but a shorter dosing interval is needed. Ceftibuten can be recommended with caution in ESBL producing E. coli except those producing SHV-4, SHV-5 and SHV-12 that confer resistance to it. If these enzymes are produced, fluoroquinolones or carbapenems could be considered. PMID- 15894466 TI - Reduced insulin-mediated citrate synthase activity in cultured skeletal muscle cells from patients with type 2 diabetes: evidence for an intrinsic oxidative enzyme defect. AB - In myotubes established from patients with type 2 diabetes (T2D), lipid oxidation and insulin-mediated glucose oxidation are reduced, whereas in myotubes from obese non-diabetic subjects, exposure to palmitate impairs insulin-mediated glucose oxidation. To determine the underlying mechanisms of these metabolic malfunctions, we studied mitochondrial respiration, uncoupled respiration and oxidative enzyme activities (citrate synthase (CS), 3-hydroxy-acyl-CoA dehydrogenase activity (HAD)) before and after acute exposure to insulin and/or palmitate in myotubes established from healthy lean and obese subjects and T2D patients. Basal CS activity was lower (14%) in diabetic myotubes compared with myotubes from lean controls (P=0.03). Incubation with insulin (1 microM) for 4 h increased the CS activity (26-33%) in myotubes from both lean (P=0.02) and obese controls (P<0.001), but not from diabetic subjects. Co-incubation with palmitate (0.6 mM) for 4 h abolished the stimulatory effect of insulin on CS activity in non-diabetic myotubes. No differences were detected in mitochondrial respiration and HAD activity between myotubes from non-diabetic subjects and T2D patients, and none of these measures responded to high levels of insulin and/or palmitate. These results provide evidence for an intrinsic defect in CS activity, which may play a role in the pathogenesis of T2D. Moreover, the data suggest that insulin resistance at the CS level can be induced by exposure to high free fatty acid levels. PMID- 15894467 TI - Cardiac volume overload rapidly induces oxidative stress-mediated myocyte apoptosis and hypertrophy. AB - Oxidative stress stimulates both growth and apoptosis in cardiac myocytes in vitro. We investigated the role of oxidative stress in the initial phases of cardiac remodeling induced in an animal model by volume overload. As plausible candidates for a connection between oxidative stress and cardiomyocyte apoptosis or hypertrophy, we explored the behaviour of two MAPKs, specifically JNK and ERK. At 48 h of overload, the greatest increase in oxidative stress coincided with a peak of cardiomyocyte apoptosis. This was possibly induced through the mitochondrial metabolism, as evidenced by the release of cytochrome c and a significant increase in the active forms of caspase-9 and -3, but not caspase-8. Oxidative stress markers significantly decreased at 96 h of overload, combined with a marked attenuation of apoptosis and the appearance of hypertrophy. The highest levels of JNK and the lowest levels of ERK phosphorylation were observed at 48 h of overload. Conversely, a sharp increase in ERK phosphorylation was detected at 96 h of overload coinciding with the hypertrophic response. Together these results show that oxidative stress is an early and transient event in myocardial volume overload. They suggest that oxidative stress mediates amplitude dependent apoptotic and hypertrophic responses in cardiomyocytes through the selective activation of, respectively, JNK and ERK. PMID- 15894468 TI - Testing the limits of the semantic illusion phenomenon: ERPs reveal temporary semantic change deafness in discourse comprehension. AB - In general, language comprehension is surprisingly reliable. Listeners very rapidly extract meaning from the unfolding speech signal, on a word-by-word basis, and usually successfully. Research on 'semantic illusions' however suggests that under certain conditions, people fail to notice that the linguistic input simply doesn't make sense. In the current event-related brain potentials (ERP) study, we examined whether listeners would, under such conditions, spontaneously detect an anomaly in which a human character central to the story at hand (e.g., "a tourist") was suddenly replaced by an inanimate object (e.g., "a suitcase"). Because this replacement introduced a very powerful coherence break, we expected listeners to immediately notice the anomaly and generate the standard ERP effect associated with incoherent language, the N400 effect. However, instead of the standard N400 effect, anomalous words elicited a positive ERP effect from about 500-600 ms onwards. The absence of an N400 effect suggests that subjects did not immediately notice the anomaly, and that for a few hundred milliseconds the comprehension system has converged on an apparently coherent but factually incorrect interpretation. The presence of the later ERP effect indicates that subjects were processing for comprehension and did ultimately detect the anomaly. Therefore, we take the absence of a regular N400 effect as the online manifestation of a temporary semantic illusion. Our results also show that even attentive listeners sometimes fail to notice a radical change in the nature of a story character, and therefore suggest a case of short-lived 'semantic change deafness' in language comprehension. PMID- 15894469 TI - Letter processing interferes with inhibition of return: evidence for cortical involvement. AB - Inhibition of return (IOR) refers to the finding that, when the time lag between a cue and a target is prolonged, the reaction to the target, when it eventually appears, is actually slower than with no cue. This phenomenon is thought to make visual search more efficient, and it is subserved by the left inferior parietal cortex and the supramarginal gyrus bilaterally. Interestingly, the very same brain structures are also involved in letter processing. Accordingly, we asked whether the two mental processes interfere with each other when simultaneously probed. The first experiment used a typical IOR procedure, but the cue/target placeholders were either simple geometric shapes or English letters. The results show that, although IOR is approximately the same across visual fields when shape placeholders are used, it is significantly lessened in the right visual field when letters are used as cue and target placeholders. To examine if this finding was due to potential spatial frequency differences between the placeholders, a second experiment using shapes and Japanese letters was conducted, and no differences in IOR were found. The supramarginal gyrus appears to be the most likely locus for the letter-IOR interference effect because it is active bilaterally in IOR, but only in the left hemisphere during letter processing. These findings provide support for the notion that IOR is not simply due to subcortical processes but also involves processing from cortical structures. PMID- 15894470 TI - Summation of semantic priming and complex sentence comprehension in Parkinson's disease. AB - Research has suggested that the integrity of semantic processing may be compromised in Parkinson's disease (PD), which may account for difficulties in complex sentence comprehension. In order to investigate the time course and integrity of semantic activation in PD, 20 patients with PD and 23 healthy controls performed a lexical decision task based on the multi-priming paradigm. Semantic priming effects were measured across stimulus onset asynchronies of 250 ms, 600 ms, and 1200 ms. Further, PD participants performed an auditory comprehension task. The results revealed significantly different patterns of semantic priming for the PD group at the 250-ms and 1200-ms SOAs. In addition, a delayed time course of semantic activation was evident for PD patients with poor comprehension of complex sentences. These results provide further support to suggest that both automatic and controlled aspects of semantic activation may be compromised in PD. Furthermore, the results also suggest that some sentence comprehension deficits in PD may be related to a reduction in information processing speed. PMID- 15894471 TI - Paradoxical lateralization of brain potentials during imagined foot movements. AB - Though each foot is controlled primarily by the contralateral hemisphere, the event-related brain potentials preceding an overt foot movement are largest over the ipsilateral side of the head. Because such "paradoxical lateralization" results from the spatial organization of the motor homunculus, it can provide a sign of motor-cortex activation. We report paradoxical lateralization in the potentials accompanying imagined foot movements, thereby demonstrating a contribution of cortical areas directly involved in movement execution. PMID- 15894472 TI - Salt formation in solid dispersions consisting of polyacrylic acid as a carrier and three basic model compounds resulting in very high glass transition temperatures and constant dissolution properties upon storage. AB - The purpose of the study was to investigate the suitability of polyacrylic acid (PAA) as a carrier in solid dispersions, with the aim to delay crystallization of basic drugs and improve their dissolution behaviour. The physicochemical properties were investigated in order to link the physical state of some model compounds to their dissolution properties. Loperamide and two structurally related substances were selected as model compounds. Solid dispersions were prepared by spray drying. The amount of residual solvents and water was determined with gas chromatography (GCS: S: solvent) and thermogravimetric analysis (TGA). The drug loading of the dispersions was determined using high performance liquid chromatography (HPLC). ADSC (alternating or temperature modulated DSC), XRD and FT-IR-spectroscopy were used to evaluate the physical state and in vitro dissolution tests were performed to measure the dissolution properties. IR-measurements demonstrated the formation of a salt between the COOH groups of the polymer and the amino-groups of the compounds. This phenomenon results in high T(g)-values of the dispersions, suppression of crystallization of the fragment molecules during preparation and an increase of the dissolution rate. Furthermore, the stability study conducted on the dispersions with loperamide showed that both, the amorphous state of the drug and the dissolution behaviour are stable under the applied storage conditions. Hence, from the experimental results it could be concluded that PAA is a suitable carrier in the formulation of stable solid dispersions for the basic compounds that were investigated. PMID- 15894473 TI - [Neonatal consequences of benzodiazepines used during the last month of pregnancy]. AB - INTRODUCTION: Since benzodiazepines (BZD) are largely prescribed during pregnancy, the Regional Pharmacovigilance Center (RPVC) of Tours regularly deals with questions about the risk of their administration to pregnant women and the monitoring of the newborns exposed in utero to these drugs. During the third trimester, we recommend a switch in the BZD maternal treatment to oxazepam, which has an intermediary half-life and no active metabolite, and a hospitalisation of the newborn in order to monitor his respiratory rate. The purpose of our study was to evaluate the neonatal consequences of BZD used at the end of pregnancy and to analysed if our recommendations were taken into account and if they were appropriate. METHODS: From 1989 to the end of 2002, we studied the files in which women had received a BZD during the 30 days prior to delivery. We analysed maternal treatments, the outcome of pregnancy and the development of the newborn, the therapeutically attitude recommended and whether or not it was respected. RESULTS: A total of 73 files were selected. Seventy neonates were born to 73 women. The newborns were hospitalised (73%) and they developed adverse reactions possibly related to the use of BZD (51,5%) : an impregnation syndrome (42%) characterized by hypotonia and hypoventilation, and a withdrawal syndrome (20%) with tremulations as the main symptom. CONCLUSION: Considering the most frequent neonatal manifestations, hospitalization and the respiratory monitoring recommended by the RPVC seemed adequate. However, the switch to oxazepam was seldom done and its advantages should be pointed out. PMID- 15894474 TI - Targeting delivery of oligonucleotide and plasmid DNA to hepatocyte via galactosylated chitosan vector. AB - Delivery of oligonucleotide to specific cells and maintenance of its biological function are important for nucleic acid therapy. The objective of this paper is to demonstrate that galactosylated low molecular weight chitosan (gal-LMWC) is a safe and effective vector of antisense oligonucleotide (ASO) and plasmid DNA for the hepatocyte targeting delivery. Gal-LMWC has been successfully prepared and MTT cytotoxic assay shows that cytotoxicity of gal-LMWC is lower than that of high molecular weight chitosan (HMWC) and low molecular weight chitosan (LMWC) in HepG2 cells. Using a complex coacervation process, gal-LMWC can form stable nano complexes with plasmid DNA or with ASO by the electrostatic interaction. The morphometrics, particle size, and the zeta potential of gal-LMWC/ASO complexes and gal-LMWC/plasmid DNA complexes are very similar. The transfection efficiency by using gal-LMWC vector is significantly higher than that of naked DNA or naked ASO in HepG2 cells. Transfection efficiency of gal-LMWC/ASO complexes and gal LMWC/plasmid DNA complexes depends on the molar ratio of the positive chitosan amino group and the negative DNA phosphate group (N/P ratio) strongly. Inhibition experiments confirm that the enhanced transfection efficiency is due to the ASGR mediated endocytosis of the gal-LMWC/ASO complexes or gal-LMWC/DNA complexes. These results suggest that gal-LMWC can be used in gene therapy to improve the transfection efficiency in vitro and in vivo. PMID- 15894475 TI - Physical characterisation of formulations for the development of two stable freeze-dried proteins during both dried and liquid storage. AB - The development of stable freeze-dried proteins requires maintaining the physical and biological integrity of the protein as well as increasing the efficiency of the manufacturing process. Our objective was to study the effects of various excipients on both the physical characterisation and the dried and liquid stability of two proteins. Thermo-physical properties of 13 formulations were determined using both differential scanning calorimetry and freeze-drying microscopy. The antigenic activity was evaluated immediately after freeze-drying and after subsequent storage in both dried and liquid state. From the comparison between glass transition (T'g) and collapse (T coll) temperatures, we concluded that the collapse temperature was a more relevant parameter than T'g for freeze drying cycle development and optimisation. One crystalline formulation composed of 4% mannitol and 1% of sucrose protected efficiently both proteins during subsequent storage in dried state (6 months at 25 degrees C) and in liquid state (3 months at 4 degrees C after rehydration). However, the freeze-drying behaviour of this crystalline formulation remained difficult to predict and control. On the other hand, two amorphous formulations composed of 4% of maltodextrin and 0.02% of Tween 80, or 5% of BSA preserved antigenic activity during storage in dried state. The glassy character of these formulations as well as their high collapse temperature values (-9 and -12 degrees C, respectively) should allow simplification and shortening of freeze-drying process. PMID- 15894476 TI - Seasonal variations of pulmonary embolism in hospitalized patients. PMID- 15894477 TI - Severity distribution of chronic obstructive pulmonary disease (COPD) in Dutch general practice. AB - The actual burden of chronic obstructive pulmonary disease (COPD) in terms of health care use and costs strongly depends on the distribution of disease severity. For the Netherlands, the distribution of diagnosed COPD was estimated by classifying all patients with a physician diagnosis of COPD from two different sources of general practitioners (GP)-data into mild (27%), moderate (55%), severe (15%) or very severe COPD (3%) based on their post-bronchodilator FEV1% predicted, according to the GOLD-guidelines. This distribution will most likely shift to the less severe stages when under-reporting and under-diagnosis are reduced. PMID- 15894478 TI - Inspiratory muscle training in adults with chronic obstructive pulmonary disease: a systematic review. AB - The purpose of this study was to conduct a systematic review to determine the effect of inspiratory muscle training (IMT) on inspiratory muscle strength and endurance, exercise capacity, dyspnea and quality of life for adults with chronic obstructive pulmonary disease (COPD). A systematic review of the literature was conducted according the Cochrane Collaboration protocol using Medline and CINAHL. Nineteen of 274 extracted articles met the inclusion criteria and addressed comparisons of interest which included: IMT versus sham; IMT versus no intervention; low- versus high-intensity IMT; and two different modes of IMT. Thirteen meta-analyses were reported. Results indicate that targeted resistive or threshold IMT was associated with significant improvements in some outcomes of inspiratory muscle strength (PI(max) (cm H2O)) and endurance (Inspiratory Threshold Loading (kPa)), exercise capacity (Borg Scale for Respiratory Effort (modified Borg scale), Work Rate maximum (Watts)), and dyspnea (Transition Dyspnea Index), whereas IMT without a target or not using threshold training did not show improvement in these variables. There was no conclusive evidence regarding quality of life measures. IMT is effective for adults with COPD when using threshold or targeted devices that control or provide a target for training intensity. PMID- 15894479 TI - Effects of inhaled HFA beclomethasone on pulmonary function and symptoms in patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease is characterized by progressive airflow limitation and pulmonary inflammation. Inhaled corticosteroids (ICS) have been shown to be effective in the reduction of the number of exacerbations and the rate of deterioration in health status in patients with more advanced chronic obstructive pulmonary disease (COPD). Therefore current international guidelines recommend ICS for patients with severe COPD (FEV1 < 50%) with at least one exacerbation within the last year. We determined the short-term effect of the inhaled corticosteroid beclomethasone in HFA 134 formulation (Ventolair) on Health related Quality of Life (HRQOL), pulmonary function and the release of the cytokines Interleukin-10 (IL-10), Granulocyte-Macrophage Colony-stimulating Factor (GM-CSF), Interferon-gamma (IFN-gamma) and Macrophage Inflammatory Protein 1alpha (MIP-1alpha) from peripheral blood monocytes of patients with COPD (n = 11) in a 12 week double blind cross over placebo-controlled study. Baseline lung function and the St. George Respiratory Questionnaire (SGRQ) were performed at the start and the end of each treatment phase. Monocytes were separated from blood at the end of each treatment phase. The treatment with Ventolair) resulted in an increase of PEF from 4.92 to 5.53 l/s and a decrease of RV% TLC (% predicted) values from 144.52 to 131.36. Inhaled HFA beclomethasone did not affect the cytokine release of IL-10, IFN-gamma, GM-CSF and MIP-1alpha. All cytokines were measured using commercially available Enzyme Linked Immun Sorbent Assay (ELISA) kits. The symptom score of the St. George Respiratory Questionnaire significantly decreased from 55.12 to 47.77 units in the active period compared to the placebo period after the treatment with HFA beclomethasone. The present study shows that a short-term treatment with inhaled steroid beclomethasone in fine particle HFA formulation decreases the hyperinflation and improves the PEF and the COPD symptoms. PMID- 15894480 TI - Methodological improvements for measuring eicosanoids and cytokines in exhaled breath condensate. AB - BACKGROUND: Exhaled breath condensate (EBC) is simple to collect and as such a non-invasive method that has attracted substantial interest in the last few years. However, several methodological concerns have been raised and it has been difficult to reproduce results between different centres. Because of low concentrations of inflammatory markers, potential loss in the sampling system may have great influence. The aim of the present study was to se if evaporation and plastic coating could facilitate detection. METHODOLOGY: Through methodological improvements, we have now made it possible to measure EBC concentrations of eicosanoids and cytokines in our system. Due to absorbance of both fatty acid derivates and proteins to several plastics, the first step is coating of all surfaces with bovine serum albumin and Tween 20. Since several assays are sensitive to these factors, the methodology has to be standardised to avoid false results. Secondly, larger amounts of EBC have to be vacuum-dried, and thereafter resolved in the respective assay buffers. The EBCs have to be concentrated 5-10 times, depending on samples and assay sensitivity. RESULTS: Due to these improvements we can measure, for example, cysteinyl-leukotrienes, leukotriene B4, prostaglandin E and 8-isoprostane. High sensitivity assays have also made it possible to measure cytokines, for example, interleukin (IL)-1beta, IL-8 and IL 13. SUMMARY: We are aware of different results from other labs. However, it seems essential to coat and to concentrate the samples in order to achieve reliable and measurable results. PMID- 15894481 TI - WITHDRAWN: Interferometric biosensing of DNA-damaging chemicals. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 15894482 TI - Learning from collaboratives. PMID- 15894483 TI - The synchronisation of lower limb responses with a variable metronome: the effect of biomechanical constraints on timing. AB - Stepping in time with a metronome has been reported to improve pathological gait. Although there have been many studies of finger tapping synchronisation tasks with a metronome, the specific details of the influences of metronome timing on walking remain unknown. As a preliminary to studying pathological control of gait timing, we designed an experiment with four synchronisation tasks, unilateral heel tapping in sitting, bilateral heel tapping in sitting, bilateral heel tapping in standing, and stepping on the spot, in order to examine the influence of biomechanical constraints on metronome timing. These four conditions allow study of the effects of bilateral co-ordination and maintenance of balance on timing. Eight neurologically normal participants made heel tapping and stepping responses in synchrony with a metronome producing 500 ms interpulse intervals. In each trial comprising 40 intervals, one interval, selected at random between intervals 15 and 30, was lengthened or shortened, which resulted in a shift in phase of all subsequent metronome pulses. Performance measures were the speed of compensation for the phase shift, in terms of the temporal difference between the response and the metronome pulse, i.e. asynchrony, and the standard deviation of the asynchronies and interresponse intervals of steady state synchronisation. The speed of compensation decreased with increase in the demands of maintaining balance. The standard deviation varied across conditions but was not related to the compensation speed. The implications of these findings for metronome assisted gait are discussed in terms of a first-order linear correction account of synchronisation. PMID- 15894484 TI - Maximum entropy image deconvolution applied to structure determination for crystal Nd1.85Ce0.15CuO4-delta. AB - Eight [100] images from a through-focus series of tetragonal crystal Nd1.85Ce0.15CuO4-delta were transformed separately into the structure images by means of maximum entropy image deconvolution. The constructed projected structure model based on the deconvoluted image is confirmed by image simulation. It is demonstrated that the image deconvolution is still successful even when some reflections fall in the vicinity of zero cross of contrast transfer function. The effectiveness and advantages of the technique are discussed. PMID- 15894485 TI - Observations on the background spectra of four LaCl 3 Ce scintillation detectors. AB - Our laboratory has purchased four LaCl (3)(Ce) scintillation crystals over the course of the last year for evaluation as part of an environmental management research and development project. In addition to the expected content of naturally radioactive (138)La, we have found that all four of these detectors are contaminated to various degrees with alpha particle emitting nuclides that we have determined to be (227)Ac and its daughters. The impact of these radionuclides on the background spectra and, thus, on the detection sensitivity of these detectors is presented and discussed. PMID- 15894486 TI - Synphilin-1 and parkin show overlapping expression patterns in human brain and form aggresomes in response to proteasomal inhibition. AB - Lewy bodies (LBs) are the characteristic inclusions of Parkinson's disease brain but the mechanism responsible for their formation is obscure. Lewy bodies (LBs) are composed of a number of proteins of which alpha-synuclein (alpha-SYN) is a major constituent. In this study, we have investigated the distribution patterns of synphilin-1 and parkin proteins in control and sporadic PD brain tissue by immunohistochemistry (IH), immunoblotting, and immunoelectron microscopy (IEM). We demonstrate the presence of synphilin-1 and parkin in the central core of a majority of LBs using IH and IEM. Using IH, we show an overlapping distribution profile of the two proteins in central neurons. Additionally, we show sensitivity of both endogenous synphilin-1 and parkin to proteolytic dysfunction and their co localization in aggresomes formed in response to the proteasome inhibitor MG-132. We confirm that synphilin-1 and parkin are components of majority of LBs in Parkinson's disease and that both proteins are susceptible to proteasomal degradation. PMID- 15894487 TI - Cytokine release induced by killed bacteria associated with anti-IFN-gamma antibody in Shigella infection. AB - An effort was made to analyze the effect of in vitro stimulation on macrophages using killed Shigella dysenteriae type-1 (KSD1) coupled with anti-Interferon Gamma (anti-IFN-gamma) antibody. The stimulated macrophages were co-cultured with primed or non-primed T-cells from Shigella infected patients. T-cell cultures were also established by co-culturing KSD1 coupled with or without PHA stimulated macrophages. Emulsified KSD1 coupled with anti-IFN-gamma antibody was found to act as a potent immunogen, inducing the release of Th1 cytokine from primed T cells cultured in acute stage of the disease. It was observed that the levels of IFN-gamma and IL-2 production rather than IL-4 and IL-6 were increased as the disease became more severe. On comparison, the subsequent values of IL-6, IFN gamma, IL-4 and IL-2 were found to be less significant in healthy primed T-cell cultures. This was also associated with a substantial production of superoxide ions (O2-), which probably inhibits the colonization of intracellular Shigella due to the presence of anti-IFN-gamma antibodies. On the other hand KSD1 with or without PHA failed to induce such responses. The above findings reflect that in the presence of anti-IFN-gamma antibody, KSD1 acts as a potential immunogen for eliciting cellular immunity against shigellosis. PMID- 15894488 TI - Plasma free amino acid profile in cancer patients. AB - Redistribution or translocation of plasma free amino acids (PFAAs) to support visceral or tumor protein synthesis is an essential feature in cancer patients. An abnormal PFAA profile might be presented via the total reflection of cancer induced protein metabolism in tumors, skeletal muscle and the liver in cancer patients. Clinical data from 13 studies have demonstrated a cancer-related PFAA profile, especially in digestive organ cancers. The PFAA profile can differ between the early and late stages of cancer. The profile is also affected by the type of cancer. Therefore, it is postulated that a detailed analysis of the PFAA profile may serve as one of the biological markers for cancer patients. PMID- 15894489 TI - Ontogeny of postsynaptic density proteins at glutamatergic synapses. AB - In glutamatergic synapses, glutamate receptors (GluRs) associate with many other proteins involved in scaffolding and signal transduction. The ontogeny of these postsynaptic density (PSD) proteins involves changes in their composition during development, paralleling changes in GluR type and function. In the CA1 region of the hippocampus, at postnatal day 2 (P2), many synapses already have a distinct PSD. We used immunoblot analysis, subcellular fractionation, and quantitative immunogold electron microscopy to examine the distribution of PSD proteins during development of the hippocampus. Synapses at P2 contained substantial levels of NR1 and NR2B and most GluR-associated proteins, including SAP102, SynGAP, the chain of proteins from GluRs/SAP102 through GKAP/Shank/Homer and metabotropic glutamate receptors, and the adhesion factors, cadherin, catenin, neuroligin, and Nr-CAM. Development was marked by substantial decreases in NR2B and SAP102 and increases in NR2A, PSD-95, AMPA receptors, and CaMKII. Other components showed more moderate changes. PMID- 15894491 TI - Sphingolipidomics: high-throughput, structure-specific, and quantitative analysis of sphingolipids by liquid chromatography tandem mass spectrometry. AB - Sphingolipids are a highly diverse category of compounds that serve not only as components of biologic structures but also as regulators of numerous cell functions. Because so many of the sphingolipids in a biological system are bioactive and are often closely related structurally and metabolically (for example, complex sphingolipids<-->ceramide<-->sphingosine<-->sphingosine 1 phosphate), to understand the role(s) of sphingolipids in a given context one must conduct a "sphingolipidomic" analysis-i.e., a structure-specific and quantitative measurement of all of these compounds, or at least all members of a critical subset. Liquid chromatography tandem mass spectrometry (LC MS/MS) is currently the only technology with the requisite structural specificity, sensitivity, quantitative precision, and relatively high-throughput capabilities for such analyses in small samples ( approximately 10(6) cells). This review describes a series of protocols that have been developed for the relatively rapid analysis of all of the molecular species from 3-ketosphinganines through sphingomyelins and some glycosphingolipids (including all the compounds that are presently regarded as sphingolipid "second messengers") using normal- and reverse phase LC to separate isometric and isobaric species (such as glucosylceramides and galactosylceramides) in combination with triple quadrupole (for MS/MS) and hybrid quadrupole-ion trap (for MS3) mass spectrometry. Also discussed are some of the issues remaining to be resolved in the analysis of the full sphingolipidome. PMID- 15894492 TI - Dynamics of replication foci in early S phase as visualized by cross-correlation function. AB - To monitor gradual changes in the replication foci distribution during early S phase, different segments of newly synthesized DNA were visualized by immunocytochemical mapping of two consecutively incorporated deoxythymidine analogs in pulse-chase-pulse experiments in HeLa cells. The resulting dual labeled fluorescence images were evaluated using cross-correlation function (CCF) analysis. General changes of CCF shape due to image deterioration caused by blur, noise, and lateral sampling (pixel size) were also discussed. Using CCF analysis of model images simulating either random initiation of new replication foci, or the firing of new foci in close proximity to completed ones, we were able to ascribe the changes in the early S replication foci distribution to the latter mechanism. In contrast to the data published previously, we monitored the dynamics of all replication foci for up to 3 h. In addition, we showed that the replication foci dynamics is well described by random walk model, so that the average de-localization of individual foci is proportional to square root of the applied chase. PMID- 15894490 TI - Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM(TM)): application to lipid-specific membrane protein topogenesis. AB - We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition. PMID- 15894493 TI - Processing of sub-syllabic speech units in the posterior temporal lobe: an fMRI study. AB - The objective of this study was to investigate phonological processing in the brain by using sub-syllabic speech units with rapidly changing frequency spectra. We used isolated stop consonants extracted from natural speech consonant-vowel (CV) syllables, which were digitized and presented through headphones in a functional magnetic resonance imaging (fMRI) paradigm. The stop consonants were contrasted with CV syllables. In order to control for general auditory activation, we used duration- and intensity-matched noise as a third stimulus category. The subjects were seventeen right-handed, healthy male volunteers. BOLD activation responses were acquired on a 1.5-T MR scanner. The auditory stimuli were presented through MR compatible headphones, using an fMRI paradigm with clustered volume acquisition and 12 s repetition time. The consonant vs. noise comparison resulted in unilateral left lateralized activation in the posterior part of the middle temporal gyrus and superior temporal sulcus (MTG/STS). The CV syllable vs. noise comparison resulted in bilateral activation in the same regions, with a leftward asymmetry. The reversed comparisons, i.e., noise vs. speech stimuli, resulted in right hemisphere activation in the supramarginal and superior temporal gyrus, as well as right prefrontal activation. Since the consonant stimuli are unlikely to have activated a semantic-lexical processing system, it seems reasonable to assume that the MTG/STS activation represents phonetic/phonological processing. This may involve the processing of both spectral and temporal features considered important for phonetic encoding. PMID- 15894494 TI - The brain uses single-trial multisensory memories to discriminate without awareness. AB - Multisensory experiences enhance perceptions and facilitate memory retrieval processes, even when only unisensory information is available for accessing such memories. Using fMRI, we identified human brain regions involved in discriminating visual stimuli according to past multisensory vs. unisensory experiences. Subjects performed a completely orthogonal task, discriminating repeated from initial image presentations intermixed within a continuous recognition task. Half of initial presentations were multisensory, and all repetitions were exclusively visual. Despite only single-trial exposures to initial image presentations, accuracy in indicating image repetitions was significantly improved by past auditory-visual multisensory experiences over images only encountered visually. Similarly, regions within the lateral-occipital complex-areas typically associated with visual object recognition processes-were more active to visual stimuli with multisensory than unisensory pasts. Additional differential responses were observed in the anterior cingulate and frontal cortices. Multisensory experiences are registered by the brain even when of no immediate behavioral relevance and can be used to categorize memories. These data reveal the functional efficacy of multisensory processing. PMID- 15894495 TI - Association between heterozygosity for HFE gene mutations and hepatitis viruses in hepatocellular carcinoma. AB - Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are strong and independent risk factors for hepatocellular carcinoma (HCC) development. Patients with hereditary hemochromatosis (HH) are considered at risk of developing cancer. However, the interaction between HFE gene mutations and hepatitis viruses for HCC development has not been systematically searched for. To assess the interaction between HFE gene mutations and exogenous risk factors in the risk of HCC occurrence, a case-only approach, in which just a series of patients is enrolled, was used. Three hundred three cirrhotic patients (231 males, 72 females) from five liver units in different geographic areas of Italy, who developed HCC during regular follow-up between January 1999 and March 2003, and whose blood DNA was available, were analyzed. In all subjects, hepatitis B surface antigen (HBsAg), anti-HCV and HFE gene mutations were assayed; alcohol intake was recorded by history. The interaction between HFE genotypes and hepatitis viruses for HCC was estimated by multivariate analysis adjusting for the confounding effect of alcohol intake, area of residence and months of follow-up. Of the 303 HCC cases, 12 (4.0%) were heterozygous for the C282Y mutation, 93 (30.7%) for the H63D, and 198 (65.3%) homozygous for the wild allele. Multivariate analysis showed that C282Y heterozygous males were 3.8-fold (95% CI=1.0-15.2) more likely to be HBV positive and that H63D heterozygous females were 6.0-fold (95% CI=1.2-113.8) more likely to be HCV positive than wild type subjects. In conclusion, given the association between C282Y mutation and HBV infection in male patients with HCC, a careful evaluation and follow-up should be considered in the C282Y-positive subjects with hepatitis B virus related liver disease. The interaction between the H63D mutation and HCV, observed only in women, may reflect a higher sensitivity to H63D-induced iron metabolism abnormalities and a reduced antioxidant capability in the presence of an even minor increase of iron which may occur as a consequence of the coexistence of hepatitis C infection and heterozygosity for HH. PMID- 15894496 TI - What sense lies in antisense inhibition of inducible nitric oxide synthase expression? AB - The impact of nitric oxide (NO) synthesized after activation by proinflammatory cytokines and/or bacterial products by an inducible NO synthase (iNOS) is still contradictory. Expression of iNOS in inflammatory reactions is often found predominantly in cells of epithelial origin, and in these cases NO may serve as a protective agent limiting pathogen spreading, downregulating local inflammatory reactions by inducing production of Th2-like responses in a classical feedback circle, or limiting tissue damage during stress conditions. However, an abundant amount of data on chronic human disorders with predominant proinflammatory Th1 like reactions points to a destructive role of iNOS activity calling for a specific inhibition. Various methods to inhibit iNOS have been established to elucidate a protective versus a destructive role of NO during various stresses. In this review, we focus on antisense (AS)-mediated gene knock-down as a relatively new method to inhibit NO production and summarize the techniques applied and their successes. At least in theory, it provides a specific, rapid, and potentially high-throughput method for inhibiting gene expression and function. We here discuss the opportunities of iNOS-directed AS-ODN, and extensively deal with limitations and experimental problems. PMID- 15894497 TI - Seasonal variation in plasma thyroxine concentrations in juvenile alligators (Alligators mississippiensis) from three Florida Lakes. AB - Circulating concentrations of thyroxine (T(4)) vary seasonally in many vertebrates. This study examined the seasonal variation in plasma concentrations of T(4) in juvenile American alligators (Alligator mississippiensis) from three populations in central Florida, USA. One site, Lake Woodruff National Wildlife Refuge, is considered a reference site whereas the other two lakes, Lake Apopka and Orange Lake, are significantly impacted by human activity. Juvenile American alligators ranging from 75-150 cm in total length were hand-captured at night from November 2000-April 2002. Plasma thyroxine concentrations were analyzed using a radioimmunoassay (RIA) previously validated for alligator plasma. Juvenile American alligators display seasonal variation in circulating T(4) concentrations. Plasma T(4) concentrations decrease from August/September to November and then begin a slow rise until April, at which point they plateau. Sex of juveniles influenced plasma concentrations of T(4) in some months but did not appear to alter the pattern in seasonal variation. The pattern we observed in plasma T(4) concentrations is not directly related to an environmental factor such as ambient temperature but is similar to that seen in plasma sex steroid concentrations during the reproductive cycle of adult alligators. Although the pattern and plasma concentration of T(4) exhibits significant variation among the three lakes studied, the pattern in seasonal variation appears similar. Comparing the seasonal pattern in plasma T(4) with plasma concentrations of sex steroids (testosterone and estradiol-17beta) or corticosterone could provide important information on the peripubescent life stage of the American alligator. PMID- 15894498 TI - [Blood transfusion practice]. AB - The publication of the Circular DGS/DHO/Afssaps N(o) 2003-582 of the December 15 2003 relating to the administration of blood and blood components, means that an update of statutory requirements is needed. This article presents the overall process from the writing of prescriptions to the care and monitoring of the transfused patient. An update and a critical analysis of regulations currently in force are presented for each step as well as a description of the main adverse events. PMID- 15894499 TI - [Towards bacterial detection in labile blood products]. AB - Bacterial contamination of blood components represents today the highest infectious risk in blood transfusion, the risk is particularly high when it affects platelet concentrates. The prevention methods developed over the past ten years (donor selection, phlebotomy site preparation, first 30 ml diversion, systematic leuko-reduction...) which aimed at limiting the introduction of bacteria in donations and bacterial proliferation, has reduced the risk of transfusion reaction due to the bacterial contamination. Improving strategies for reducing the risks of bacterial contamination is one of the priorities of the French National Blood Transfusion Service (l'Etablissement francais du sang - EFS). It is essential to improve existent prevention methods and develop the implication of all the actors (from donation to transfusion) involved in the prevention of this risk. Bacterial detection or pathogens inactivation are now available and are able to reduce (for detection) or prevent (for inactivation) the occurrence of reaction due to bacterial contamination of PC. Up to now, the choice is in favour of bacterial detection. Three methods (BacT/Alert, BioMerieux; eBDS, Pall; ScanSystem, Hemosystem) of detection of bacterial contamination in PC can be generalised now. Adaptations, need for their implementation are acceptable, especially concerning PC availability. PMID- 15894500 TI - [Electronic learning: interactive learning in medicine or Socrates in electronic guise]. AB - E-learning has been widely used for training in different fields. More recently, it was introduced during medical studies or for continuous medical education. The Canadian Universities are pioneers in e-learning creating special departments dedicated to pedagogy. Developing countries like Brazil or Central Europe have made some pilot experiments, which were successful. Several electronic companies have given a free access to the programmes and sites. The use of electronic media leads to an adaptation of teaching methods making them more interactive. PMID- 15894501 TI - [Management and evolution of French blood establishment (EFS) laboratories' accreditation]. AB - The French Committee of Accreditation (COFRAC) is the independent organism, internationally acknowledged, that authenticate the competences of the medical biology analysis laboratories, which volunteer for the accreditation. The main EFS laboratories are candidates for this recognition of their competences by the Cofrac, according to the ISO/CEI 17025 standard and its documents of application. A national project at horizon 2009 has been elaborated, involving internal resources from every part of the establishment. This is a constructive approach lays the foundations of a methodology of initial and continuous control of analysis methods, which meets the automation and computerization requirements. It highlights gaps as many as new orientations opening the way at for developments and a concerted harmonization of the practices harmonization. PMID- 15894502 TI - [Indications for use of labile blood products and the physiology of blood transfusion in medicine. The French Agency for the Health Safety of Health Products]. AB - This article presents the French national recommendations for the use of blood products in medicine. PMID- 15894503 TI - [The BOTIA project ("Blood and Organ Transmissible Infectious Agents"): a European collection of blood samples and an observatory of agents transmitted by blood transfusion or organ transplantation]. PMID- 15894504 TI - [Incidents of blood donation]. AB - In France, more than two million blood donations are needed for transfusion each year. The rules and requirements for blood donors selection are strict and aim to protect the donors' health as well as the patients'security. However, the review of the literature shows that adverse effects of blood withdrawal occur quite frequently. They can be classified in immediate or delayed events, or local (hematoma, nerve injury, arterial puncture injury, allergy, veinitis, thrombosis) and general reactions (vasovagal reaction, citrate toxicity, cardiovascular failure, allergy to ethylene oxide, adverse effects of drugs used for some types of donation). This article reviews the principal adverse events described in the literature and concludes with the necessity to set up hemovigilance for donors, as defined in directive 2002/98/EC of the European parliament and of the council. PMID- 15894505 TI - [Cellular mechanisms implicated in anti-erythrocyte alloimmunization]. AB - In many clinical situations patients are dependent on blood transfusions. Occurrence of alloimmunization to blood group antigens (BGA) complicates the transfusion strategy and may be involved in clinical transfusion stalemate situations. B cell differentiation into antibody-secreting plasma cells is triggered by antigen and requires helper T cells which produce cytokines. Although antibodies implicated in BGA alloimmunization have been studied for many years, little is known about helper T cell responses that drive their production. Few studies on BGA specific T cell responses have been published today. This review summarizes the new developments in the field of cellular mechanisms implicated into antibody production. The definition of immunodominant peptides derived from RhD and Jk(a) BGAs, the cytokine patterns induced and the HLA class II molecules implicated in their presentation are analyzed. A tolerogenic route for RhD immunodominant peptides is experimented. Identification of such immunodominant peptides, the cytokine patterns induced and the HLA class II molecules implicated in their presentation, would facilitate the design of new therapeutic strategies including the specific control of alloimmunization with peptide antigen tolerogens or the ex-vivo induction of regulatory T cells. PMID- 15894506 TI - [Bacterial detection leading to pathogen inactivation]. AB - Bacterial contamination of blood components remains the highest infectious risk in blood transfusion, the risk is particularly high when it affects platelet concentrates (PC). In France, the residual risk of transfusion reaction due to bacterial contamination of PC has been decreasing slowly since 1994 but for all severity 1 case occurs with about 25,000 distributed PC and one death occurs with 200,000 distributed units. This reduction of the risk may be due to the measures which were implemented during the last 10 years in order to prevent contamination during donation. Improving strategies for reducing the risks of bacterial contamination is one of the priorities of the French National Blood Transfusion Service (l'Etablissement Francais du sang - EFS). The main target remains PC. Bacterial detection or pathogens inactivation are now available and are able to reduce (for detection) or prevent (for inactivation) the occurrence of reaction due to bacterial contamination of PC. Up to now, the choice is in favour of bacterial detection. A national study was carried out in seven regional EFS at the end of 2004. It aims at confirming the feasibility of a systematic bacterial screening of PC before their delivery. The first conclusions show that this screening can be implemented with acceptable modifications in term of platelets availability. We can expect in a next future that new pathogens reduction technique and/or new detection systems will be available, certainly more efficient to prevent reaction due to bacterial contamination. Implementation of actual detection methods is probably a temporary solution. PMID- 15894507 TI - [Equipment qualification]. AB - The qualification of the equipment is a particularly important stage in the transfusional process. On the one hand, of many standards such as those of certification or that of accreditation require it, just as the good transfusional practices; in addition, the practices of steps of quality assurance develop this aspect. Indeed, the absence of the realization of this qualification of material having an influence on the finished product, can lead to an error in the product. This qualification passes by various stages of which some are major such as the drafting of the schedule of conditions, the drafting of the operational protocol of qualification, the decision made for the setting in routine. Finally so that this qualification takes all its dimensions it is necessary to carry out methods linked to the international system of measurement. Moreover certain questions after reflexions must find response such as which unit to check, and only this one, the equipment is - it a complex one, is there a maintenance contract? Once all these elements taken into account, the questions having found their answer, the operational protocol will then well be built, the decisions of settings in routine could be done and the sets of the finalized stages. PMID- 15894508 TI - [TRALI: from diagnosis to prevention]. AB - Transfusion-Related Acute Lung Injury (TRALI) is a post-transfusion acute respiratory distress syndrome (ARDS). TRALI is a non-cardiogenic lung oedema occurring within 6 hours following the infusion of a blood component. Its frequency has been estimated from 1 in 5.000 to 1 in 500.000 injected blood products. Its aetiology is still controversial. Antibodies against HLA class I molecules or granulocyte surface molecules recognizing recipient leukocyte antigens were implicated at first. More recently anti HLA class II were involved in some cases. Finally granulocyte activating lipids released from cells during blood storage were claimed to account for cases in which no antibody were detected. Nevertheless, in most cases, none of these triggering factors alone seems sufficient to induce a TRALI. A predisposing condition, associated with leucostasis in pulmonary vessels, is required. Whatever the mechanism, the pulmonary lesion is eventually due to release of neutrophil granule content in contact with endothelial cells of lung micro-capillary vessels. The basement membrane damages leads to fluid and cell extravasations in interstitial and alveolar spaces. Among blood donors multiparous women are the most frequently involved. Up to now there are neither definite guidelines regarding detection of harmful antibodies nor regulation for deferral of potentially dangerous blood donors. PMID- 15894509 TI - [Evolution of the practices of labile blood product distribution]. PMID- 15894510 TI - [Haematopoietic stem cell specimens]. PMID- 15894511 TI - Localization of anatomical point landmarks in 3D medical images by fitting 3D parametric intensity models. AB - We introduce a new approach for the localization of 3D anatomical point landmarks. This approach is based on 3D parametric intensity models which are directly fitted to 3D images. To efficiently model tip-like, saddle-like, and sphere-like anatomical structures we introduce analytic intensity models based on the Gaussian error function in conjunction with 3D rigid transformations as well as deformations. To select a suitable size of the region-of-interest (ROI) where model fitting is performed, we also propose a new scheme for automatic selection of an optimal 3D ROI size based on the dominant gradient direction. In addition, to achieve a higher level of automation we present an algorithm for automatic initialization of the model parameters. Our approach has been successfully applied to accurately localize anatomical landmarks in 3D synthetic data as well as 3D MR and 3D CT image data. We have also compared the experimental results with the results of a previously proposed 3D differential approach. It turns out that the new approach significantly improves the localization accuracy. PMID- 15894512 TI - Modifications of DNA by platinum complexes. Relation to resistance of tumors to platinum antitumor drugs. AB - The importance of platinum drugs in cancer chemotherapy is underscored by the clinical success of cisplatin [cis-diamminedichloroplatinum(II)] and its analogues and by clinical trials of other, less toxic platinum complexes that are active against resistant tumors. The antitumor effect of platinum complexes is believed to result from their ability to form various types of adducts with DNA. Nevertheless, drug resistance can occur by several ways: increased drug efflux, drug inactivation, alterations in drug target, processing of drug-induced damage, and evasion of apoptosis. This review focuses on mechanisms of resistance and sensitivity of tumors to conventional cisplatin associated with DNA modifications. We also discuss molecular mechanisms underlying resistance and sensitivity of tumors to the new platinum compounds synthesized with the goal to overcome resistance of tumors to established platinum drugs. Importantly, a number of new platinum compounds were designed to test the hypothesis that there is a correlation between the extent of resistance of tumors to these agents and their ability to induce a certain kind of damage or conformational change in DNA. Hence, information on DNA-binding modes, as well as recognition and repair of DNA damage is discussed, since this information may be exploited for improved structure-activity relationships. PMID- 15894513 TI - Increased prevalence of transfusion-transmitted virus and cross-reactivity with immunodominant epitopes of the HRES-1/p28 endogenous retroviral autoantigen in patients with systemic lupus erythematosus. AB - OBJECTIVE: Systemic lupus erythematosus (SLE) patients produce autoantibodies to HRES-1/p28, a human endogenous retrovirus-encoded nuclear protein. To identify cross-reactive viral antigens capable of triggering autoreactivity, HRES-1/p28 epitopes were mapped by SLE antibodies. METHODS: Forty-four peptides overlapping HRES-1/p28 and 13 viral peptides were synthesized on cellulose membrane and tested for recognition by antibodies from 16 HRES-1 Western blot seropositive SLE patients. Transfusion-transmitted virus (TTV) was detected by gene amplification in sera of 211 SLE patients, 78 healthy SLE family members, 199 unrelated healthy donors, and 91 rheumatoid arthritis (RA) patients. RESULTS: HRES-1/p28 residues 41-55, 121-135, and 156-170 were recognized by 12/16 (75.0%), 11/16 (68.8%), and 9/16 lupus sera (56.25%) and considered immunodominant. HRES-1/p28 residues 121 135 harbor cross-reactive epitope with retroviral peptides and the 70 K U1snRNP lupus autoantigen. HRES-1/p28 residues 41-55 and 156-170 exhibited the highest prevalence of cross-reactivity with TTV peptide ORF2a (14/16, 87%). Prevalence of TTV DNA was increased in lupus patients (120/211) with respect to healthy (66/199; P < 0.0001) or RA controls (23/91; P < 0.0001). TTV prevalence in healthy lupus relatives (40/78) was decreased with respect to lupus patients (80/121; P = 0.0184) and increased with respect to unrelated healthy donors (66/199; P = 0.0026). HRES-1/p28 Western blot reactivity was observed in 12/23 TTV PCR-negative donors and 43/58 TTV PCR-positive donors (P < 0.0281). CONCLUSIONS: Increased prevalence of TTV and molecular mimicry with HRES-1/p28 may contribute to generation of antinuclear antibodies and pathogenesis of SLE. PMID- 15894514 TI - Preserved systolic function with isolated left ventricular noncompaction in an elderly patient. AB - Isolated left ventricular noncompaction (IVNC) is a rare congenital cardiomyopathy resulting from an arrest in normal endomyocardial embryogenesis. Clinical presentations of IVNC include systolic and diastolic dysfunction, systemic embolism and ventricular arrhythmias. In recent years there has been an increasing awareness of this anomaly; however, especially in elderly, clinical characterization and natural course of IVNC are still in question. In this case, we report a case of a 78-year-old patient with the diagnosis of IVNC with preserved left ventricular (LV) systolic function. To the best of our knowledge, this is the oldest IVNC case with preserved LV systolic function in the literature. PMID- 15894515 TI - Interest of tumor necrosis factor-alpha -308 G/A and interleukin-10 -592 C/A polymorphisms in human African trypanosomiasis. AB - This study aimed to determine whether single nucleotide polymorphisms (SNPs) within tumour necrosis factor-alpha (TNF) and interleukin-10 (IL10) promoters and genes are associated with human African trypanosomiasis (HAT). The polymorphisms used in the analysis were TNF(-308G/A), TNF(-238G/A), TNF(-1031T/C), TNF(+488G/A), IL10(-1082G/A) and IL10(-592C/A). A familial case-control sample of 277 individuals (102 cases and 175 parents) and a matched case-control group of 225 subjects (88 cases and 137 unrelated controls) were gathered together in this study. A conditional logistic regression was used to test for association. We carried out this analysis in the overall population and after stratification by time of exposure, age and ethnic group. Our results show that in the overall population, and after stratification by time of exposure, the IL10(-592A) allele is associated with a lower risk of disease, suggesting the possibility of a protective effect. After stratification by time of exposure, individuals homozygous for the SNP located in the TNF(-308) promoter were shown to present a higher risk of developing the disease early after exposure. Our study shows that TNF(-308G/A) and IL10(-592C/A) SNPs are polymorphisms of interest in the investigation of the genetic susceptibility to human African trypanosomiasis. Larger studies are currently underway to confirm these results. PMID- 15894516 TI - Neonatal maternal separation and early life programming of the hypoxic ventilatory response in rats. AB - The neonatal period is critical for central nervous system (CNS) development. Recent studies have shown that this basic neurobiological principle also applies to the neural circuits regulating respiratory activity as exposure to excessive or insufficient chemosensory stimuli during early life can have long-lasting consequences on the performance of this vital system. Although the tactile, olfactory, and auditory stimuli that the mother provides to her offspring during the neonatal period are not directly relevant to respiratory homeostasis, they likely contribute to respiratory control development. This review outlines the rationale for the link between maternal stimuli and programming of the hypoxic ventilatory response during early life, and presents recent results obtained in rats indicating that experimental disruption of mother-pup interaction during this critical period elicits significant phenotypic plasticity of the hypoxic ventilatory response. PMID- 15894518 TI - Simultaneous retention index analysis of urinary amino acids and carboxylic acids for graphic recognition of abnormal state. AB - Simultaneous profiling analysis of urinary amino acids (AAs) and carboxylic acids (CAs) was combined with retention index (I) analysis for graphic recognition of abnormal metabolic state. The temperature-programmed I values of the AA and CA standards measured as ethoxycarbonyl (EOC)/methoxime (MO)/tert-butyldimethylsilyl (TBDMS) derivatives were used as the reference I values. Urine samples were subjected to the sequential EOC, MO and TBDMS reactions for the analysis by gas chromatography (GC) and GC-mass spectrometry. The complex GC profiles were then transformed into their respective I patterns in bar graphic forms by plotting the normalized peak area ratios (%) of the identified AAs and CAs against their reference I values as the identification numbers. When the present method was applied to infant urine specimens from normal controls and patients with inherited metabolic diseases such as phenylketonuria, maple syrup urine disease, methylmalonic aciduria or isovaleric aciduria, each I pattern of bar graph more distinctly displayed quantitative abundances of urinary AAs and CAs in qualitative I scale, thus allowing graphic discrimination between normal and abnormal states. PMID- 15894519 TI - Features of the acid protease partition in aqueous two-phase systems of polyethylene glycol-phosphate: chymosin and pepsin. AB - The partitioning of chymosin (from Aspergilus niger) and pepsin (from bovine stomach) was carried out in aqueous-two phase systems formed by polyethyleneglycol-potassium phosphate. The effects of polymer concentration, molecular mass and temperature were analysed. The partition was assayed at pH 7.0 in systems of polyethyleneglycol of molecular mass: 1450, 3350, 6000 and 8000. Both proteins showed high affinity for the polyethyleneglycol rich phase. The increase of polyethyleneglycol concentration favoured the protein transfer to the top phase, suggesting an important protein-polymer interaction. Polyethyleneglycol proved to have a stabilizing effect on the chymosin and pepsin, increasing its protein secondary structure. This finding agreed with the enhancement of the milk clotting activity by the polyethyleneglycol. The method appears to be suitable as a first step for the purification of these proteins from their natural sources. PMID- 15894520 TI - Bilirubin removal from human plasma by Cibacron Blue F3GA using immobilized microporous affinity membranous capillary method. AB - A novel affinity sorbent system for direct bilirubin removal from human plasma was developed. These new adsorbents comprise Cibacron Blue F3GA as the specific ligand, and microporous membranous poly(tetrafluoroethylene) capillary (modified by coating with a hydrophilic layer of poly(vinyl alcohol) after activation) as the carrier matrix. The affinity adsorbents carrying 126.5 micromol Cibacron Blue F3GA/g polymer was then used to remove bilirubin in a flow-injection system. Non specific adsorption on the poly(vinyl alcohol) coated capillary remains low, and higher affinity adsorption capacity, of up to 76.2 mg/g polymer was obtained after dye immobilization. The bilirubin adsorption capacity of the affinity capillary decreased with increase in the recirculation rate of plasma. The adsorption capacity increased with increase the temperature while decreased with increase the ionic strength. The maximum adsorption was only observed in neutral solution (pH 6-7). The adsorption isotherm fitted the Langmuir model well. These new adsorbents have higher velocity of mass transfer, better adsorption capacity, less fouling, longer service life and good reusability. The results of blood tests suggested the dye affinity capillary has good blood compatibility. PMID- 15894521 TI - Stability, pKa and plasma protein binding of roscovitine. AB - In the present investigation, the binding of roscovitine (100, 500 and 1500 ng/mL) to plasma proteins was studied at 25 and 37 degrees C by ultrafiltration and equilibrium dialysis methods. Drug stability in plasma was assessed during a 48 h at 4, 25 and 37 degrees C. The effect of thawing and freezing on drug stability was studied. The pKa of roscovitine was measured using capillary electrophoresis coupled with mass spectrometry. Roscovitine was quantified utilizing liquid chromatography and tandem mass spectrometry. Roscovitine is highly bound to plasma proteins (90%). Binding of roscovitine to human serum albumin was constant (about 90%) within concentration range studied while the binding to alpha1-acid glycoprotein decreased with increasing drug concentration indicating that albumin is more important in clinical settings. However, alpha1 acid glycoprotein might be important when plasma proteins change with disease. Protein binding was higher at 25 degrees C compared to 37 degrees C. The results obtained by equilibrium dialysis were in good agreement with those obtained by ultrafiltration. Roscovitine was stable at all temperatures studied during 48 h. Roscovitine has a pKa of 4.4 showing that the drug mainly acts like a weak mono base. The results obtained in our studies are important prior to clinical trials and to perform pharmacokinetic studies. PMID- 15894522 TI - Love, careers, and heights in France, 2001. AB - Short men are less likely to be married or live in a permanent relationship than their taller counterparts. This pattern is not due to their social status. While blue-collar workers are shorter on average than managers, the effects of height on finding a mate are similar in the two social groups. Being tall is also economically advantageous for men. With identical educational attainment levels, tall men have better careers than short men as they are given greater supervisory responsibilities. In making a commitment, some women might take height into account as an anticipated indicator of future resources of the household. Choice of partner is also influenced by social norms--i.e., partners should be physically well-matched--which is more difficult for shorter men. PMID- 15894523 TI - Gene regulation and physiological function of placental leucine aminopeptidase/oxytocinase during pregnancy. AB - Human pregnancy serum and placenta have the ability to degrade uterotonic peptide oxytocin (OT). Placental leucine aminopeptidase (P-LAP), which is also called cystine aminopeptidase, is the only membrane aminopeptidase known to functionally degrade OT as oxytocinase (OTase). P-LAP/OTase hydrolyzes several peptides other than OT including vasopressin and angiotensin III. P-LAP/OTase predicted from cDNA sequence is a type II integral membrane protein, which is converted to a soluble form existing in maternal serum by metalloproteases, possibly ADAM (a disintegrin and metalloproteinase) members. P-LAP/OTase activity increases with normal gestation, while decreases in the patients with preterm delivery and severe preeclampsia. In placenta, P-LAP/OTase is predominantly expressed in differentiated trophoblasts, syncytiotrophoblasts. Activator protein-2 (AP-2) and Ikaros transcription factors play significant roles in exerting high promoter activity of P-LAP/OTase in the trophoblastic cells. Moreover, P-LAP/OTase is transcriptionally regulated in a trophoblast-differentiation-dependent fashion via up-regulation of AP-2, putatively AP-2alpha. P-LAP/OTase may be involved in maintaining pregnancy homeostasis via metabolizing peptides such as OT and vasopressin. PMID- 15894524 TI - Crystallization and preliminary crystallographic analysis of Bacillus thuringiensis AHL-lactonase. AB - The quorum sensing (QS) systems in Gram-negative bacteria are mostly associated with diffusible N-acyl-L-homoserine lactones (AHLs). AHL-degrading enzymes hydrolyze the AHLs into inactive molecules, thereby blocking the QS systems that are closely linked to virulence factor production and biofilm formation. Consequently, these enzymes have recently attracted intense interest for the development of anti-infection therapies for plants and animals. However, despite significant progress in the investigation of AHL-degrading enzymes, no structure is yet available. Accordingly, this study reports on the expression and purification of the AHL-lactonase from Bacillus thuringiensis subsp. kurstaki HD263, as well as the successful crystallization of the enzyme. High-quality native crystals were obtained and a complete data set collected at 2.0 A resolution. The native crystal was found to belong to the space group P2(1)2(1)2(1), with unit cell parameters a=52.7 A, b=55.9 A, and c=74.1 A and one molecule in the asymmetric unit. MAD data were also collected at 2.4 A resolution for a SeMet-substituted crystal. PMID- 15894525 TI - Enhanced tumor regression and tissue repair when zoledronic acid is combined with ifosfamide in rat osteosarcoma. AB - The efficacy of zoledronic acid (ZOL), with or without the anticancer drug ifosfamide (IFO), was tested on primary bone tumor growth using a rat transplantable model of osteosarcoma. The effects on bone remodeling and tumor growth were analyzed by radiography, micro-computed tomography (micro-CT), and histological staining. The in vitro effects of ZOL were studied by proliferation, apoptosis, and cell cycle analyses on the osteosarcoma cells OSRGA compared to rat primary osteoblasts. Treatment with ZOL was effective in preventing the formation of osteolytic lesions that developed in bone sites and in reducing the local tumor growth, as compared to the untreated rats. The combination of ZOL and IFO was more effective than each agent alone in preventing tumor recurrence, improving tissue repair, and increasing bone formation as revealed by the analysis of trabecular architecture. In vitro studies demonstrated that ZOL was more potent against the OSRGA cell line than osteoblasts (with a half-maximal inhibitory effect on proliferation seen at 0.2 and 20 microM, respectively), the ZOL-induced inhibition of OSRGA proliferation being due to cell cycle arrest in S phase. No effect on OSRGA apoptosis could be observed in vitro, as assessed by Hoechst staining and caspase-1 and -3 activation. In situ cell death was determined by TUNEL staining on tumor tissue sections. No significant difference in TUNEL-positive cells could be observed between ZOL-treated and -untreated rats. This is the first report of the anti-bone resorption and antitumoral activities of zoledronic acid in a rat model of osteosarcoma, and its beneficial association with an antitumoral chemotherapeutic drug in preventing tumor recurrence. PMID- 15894526 TI - Deletion of iNOS gene impairs mouse fracture healing. AB - Nitric oxide (NO) is a signaling molecule synthesized from l-arginine by nitric oxide synthases (NOSs). NOS isoforms are either constitutive (endothelial NOS [eNOS] and neuronal NOS [nNOS]) or inducible NOS (iNOS). Previously, our group has reported that NO is expressed during and modulates fracture healing. In this study, we evaluated the specific contribution of iNOS to fracture healing by using iNOS gene therapy in iNOS-deficient mice. Twelve-week-old female wild-type mice and iNOS-KO mice had a right femoral midshaft osteotomy fixed with an intramedullary 0.5-mm-diameter needle. A gelatine sponge was implanted across the fracture site. The gelatine sponge received either Ad5-CMViNOS (in iNOS-deficient mice; n=16) or Ad5-CMVempty (in wild-type mice; n=15, and iNOS-deficient mice; n=15) at a dose of 10(7) pfu. Mice were sacrificed at day 14, and their right and left hind limbs were harvested. Cross-sectional area (CSA) was determined by measuring the callus diameter across the mediolateral and anteroposterior plane using a vernier caliper. Specimens were loaded to failure torsionally in a biaxial INSTRON testing system, and maximum torque, torsional stiffness, and maximal and total energy were determined. Deletion of the iNOS gene decreased the total and maximum energy absorption of the healing femoral fracture by 30% and by 70% (P<0.01), respectively, in comparison to the wild-type mice. This reduction in energy absorption was reversed by iNOScDNA administration via adenovirus vector. Furthermore, iNOScDNA caused an increase in torsional failure by 20% (P=0.01) in comparison to iNOS(-/-) mice that did not receive the iNOScDNA. There were no significant differences in the biomechanical properties of intact femora. These data indicate that iNOS is important in mouse fracture healing. However, the clinical utility of NOS gene therapy to enhance fracture healing will need further evaluation. PMID- 15894527 TI - The effect of risedronate on bone mineralization as measured by micro-computed tomography with synchrotron radiation: correlation to histomorphometric indices of turnover. AB - The primary goal of our study was to determine changes in bone mineralization in postmenopausal osteoporotic women treated for 3 years with risedronate or placebo. A secondary goal was to determine the relationship between mineralization and indices of bone turnover measured on the same biopsies. The degree of mineralization was measured by micro-computed tomography using Synchrotron radiation (Synchrotron microCT) in the trabecular bone of paired transiliac biopsies taken at baseline and after 3 years of treatment from patients receiving risedronate 5 mg daily (n=11) or placebo (n=8). In the risedronate-treated patients, the average mineralization (Avg-MIN) and peak mineralization (Peak-MIN) at 3 years were significantly increased from baseline by 4.7% (P<0.0001) and 5.4% (P=0.0003), respectively and showed significant negative correlation to turnover indices. In the placebo-treated patients, the increases in Avg-MIN (2.0%) and Peak-MIN (1.6%) were not significantly different from baseline and correlation to turnover indices was weaker. Risedronate significantly reduced the ratio of low- to high-mineralized bone fractions estimated by volume (BMR-V) and surface area (BMR-S) by 70.1% and 54.1%, respectively from baseline. These changes were consistent with the significant reduction of turnover from baseline assessed by reductions in mineralizing surface, MS/BS (-72.8%); activation frequency, Ac.F (-60.4%); and bone formation rate, BFR-BV (-63.6%) in the same biopsies in the risedronate-treated patients. Comparing the pair-wise changes from baseline, risedronate significantly reduced the low-mineralized bone fraction in comparison to placebo, as indicated by a larger reduction of BMR-V (P=0.015) and BMR-S (P=0.035). In the risedronate group, BMR-V and BMR-S showed significant positive correlation to MS/BS (R2: 0.83 and 0.92, respectively). The correlations to Ac.F and BFR-BV were also significant, with BMR-S showing a strong relation (R2: 0.77 and 0.79, respectively). The data suggest that BMR-V and BMR-S are markers of turnover of trabecular bone and may be used to assess treatment effect on turnover in bone biopsies. The results demonstrate that the reduction of turnover by risedronate increased the degree of mineralization and reduced the ratio of low- to high mineralized bone fractions which may increase bone's resistance to fracture. PMID- 15894528 TI - Titan's atmospheric temperatures, winds, and composition. AB - Temperatures obtained from early Cassini infrared observations of Titan show a stratopause at an altitude of 310 kilometers (and 186 kelvin at 15 degrees S). Stratospheric temperatures are coldest in the winter northern hemisphere, with zonal winds reaching 160 meters per second. The concentrations of several stratospheric organic compounds are enhanced at mid- and high northern latitudes, and the strong zonal winds may inhibit mixing between these latitudes and the rest of Titan. Above the south pole, temperatures in the stratosphere are 4 to 5 kelvin cooler than at the equator. The stratospheric mole fractions of methane and carbon monoxide are (1.6 +/- 0.5) x 10(-2) and (4.5 +/- 1.5) x 10(-5), respectively. PMID- 15894529 TI - Cassini measurements of cold plasma in the ionosphere of Titan. AB - The Cassini Radio and Plasma Wave Science (RPWS) Langmuir probe (LP) sensor observed the cold plasma environment around Titan during the first two flybys. The data show that conditions in Saturn's magnetosphere affect the structure and dynamics deep in the ionosphere of Titan. The maximum measured ionospheric electron number density reached 3800 per cubic centimeter near closest approach, and a complex chemistry was indicated. The electron temperature profiles are consistent with electron heat conduction from the hotter Titan wake. The ionospheric escape flux was estimated to be 10(25) ions per second. PMID- 15894530 TI - On the origin of leprosy. AB - Leprosy, a chronic human disease with potentially debilitating neurological consequences, results from infection with Mycobacterium leprae. This unculturable pathogen has undergone extensive reductive evolution, with half of its genome now occupied by pseudogenes. Using comparative genomics, we demonstrated that all extant cases of leprosy are attributable to a single clone whose dissemination worldwide can be retraced from analysis of very rare single-nucleotide polymorphisms. The disease seems to have originated in Eastern Africa or the Near East and spread with successive human migrations. Europeans or North Africans introduced leprosy into West Africa and the Americas within the past 500 years. PMID- 15894531 TI - Benefits of early invasive treatment for acute coronary syndromes: lost in translation? PMID- 15894532 TI - Early asymmetry of gene transcription in embryonic human left and right cerebral cortex. AB - The human left and right cerebral hemispheres are anatomically and functionally asymmetric. To test whether human cortical asymmetry has a molecular basis, we studied gene expression levels between the left and right embryonic hemispheres using serial analysis of gene expression (SAGE). We identified and verified 27 differentially expressed genes, which suggests that human cortical asymmetry is accompanied by early, marked transcriptional asymmetries. LMO4 is consistently more highly expressed in the right perisylvian human cerebral cortex than in the left and is essential for cortical development in mice, suggesting that human left-right specialization reflects asymmetric cortical development at early stages. PMID- 15894533 TI - New aspects of vascular remodelling: the involvement of all vascular cell types. AB - Conventionally, the architecture of arteries is based around the close-packed smooth muscle cells and extracellular matrix. However, the adventitia and endothelium are now viewed as key players in vascular growth and repair. A new dynamic picture has emerged of blood vessels in a constant state of self maintenance. Recent work raises fundamental questions about the cellular heterogeneity of arteries and the time course and triggering of normal and pathological remodelling. A common denominator emerging in hypertensive remodelling is an early increase in adventitial cell density suggesting that adventitial cells drive remodelling and may initiate subsequent changes such as re-arrangement of smooth muscle cells and extracellular matrix. The organization of vascular smooth muscle cells follows regular arrangements that can be modelled mathematically. In hypertension, new patterns can be quantified in these terms and give insights to how structure affects function. As with smooth muscle, little is known about the organization of the vascular endothelium, or its role in vascular remodelling. Current observations suggest that there may be a close relationship between the helical organization of smooth muscle cells and the underlying pattern of endothelial cells. The function of myoendothelial connections is a topic of great current interest and may relate to the structure of the internal elastic lamina through which the connections must pass. In hypertensive remodelling this must present an organizational challenge. The objective of this paper is to show how the functions of blood vessels depend on their architecture and a continuous interaction of different cell types and extracellular proteins. PMID- 15894534 TI - Brain stem excitatory and inhibitory signaling pathways regulating bronchoconstrictive responses. AB - This review summarizes recent work on two basic processes of central nervous system (CNS) control of cholinergic outflow to the airways: 1) transmission of bronchoconstrictive signals from the airways to the airway-related vagal preganglionic neurons (AVPNs) and 2) regulation of AVPN responses to excitatory inputs by central GABAergic inhibitory pathways. In addition, the autocrine paracrine modulation of AVPNs is briefly discussed. CNS influences on the tracheobronchopulmonary system are transmitted via AVPNs, whose discharge depends on the balance between excitatory and inhibitory impulses that they receive. Alterations in this equilibrium may lead to dramatic functional changes. Recent findings indicate that excitatory signals arising from bronchopulmonary afferents and/or the peripheral chemosensory system activate second-order neurons within the nucleus of the solitary tract (NTS), via a glutamate-AMPA signaling pathway. These neurons, using the same neurotransmitter-receptor unit, transmit information to the AVPNs, which in turn convey the central command to airway effector organs: smooth muscle, submucosal secretory glands, and the vasculature, through intramural ganglionic neurons. The strength and duration of reflex induced bronchoconstriction is modulated by GABAergic-inhibitory inputs and autocrine-paracrine controlling mechanisms. Downregulation of GABAergic inhibitory influences may result in a shift from inhibitory to excitatory drive that may lead to increased excitability of AVPNs, heightened airway responsiveness, and sustained narrowing of the airways. Hence a better understanding of these normal and altered central neural circuits and mechanisms could potentially improve the design of therapeutic interventions and the treatment of airway obstructive diseases. PMID- 15894535 TI - Very intense exercise-training is extremely potent and time efficient: a reminder. PMID- 15894536 TI - Metabolic energy and muscular activity required for leg swing in running. AB - The metabolic cost of leg swing in running is highly controversial. We investigated the cost of initiating and propagating leg swing at a moderate running speed and some of the muscular actions involved. We constructed an external swing assist (ESA) device that applied small anterior pulling forces to each foot during the first part of the swing phase. Subjects ran on a treadmill at 3.0 m/s normally and with ESA forces up to 4% body weight. With the greatest ESA force, net metabolic rate was 20.5% less than during normal running. Thus we infer that the metabolic cost of initiating and propagating leg swing comprises approximately 20% of the net cost of normal running. Even with the greatest ESA, mean electromyograph (mEMG) of the medial gastrocnemius and soleus muscles during later portions of stance phase did not change significantly compared with normal running, indicating that these muscles are not responsible for the initiation of leg swing. However, with ESA, rectus femoris mEMG during the early portions of swing phase was as much as 74% less than during normal running, confirming that it is responsible for the propagation of leg swing. PMID- 15894537 TI - Muscle strength response to strength training is influenced by insulin-like growth factor 1 genotype in older adults. AB - Strength training (ST) is considered an intervention of choice for the prevention and treatment of sarcopenia. Reports in the literature have suggested that the insulin-like growth factor I protein (IGF-I) plays a major role in ST-induced skeletal muscle hypertrophy and strength improvements. A microsatellite repeat in the promoter region of the IGF1 gene has been associated with IGF-I blood levels and phenotypes related to IGF-I in adult men and women. To examine the influence of this polymorphism on muscle hypertrophic and strength responses to ST, we studied 67 Caucasian men and women before and after a 10-wk single-leg knee extension ST program. One repetition maximum strength, muscle volume via computed tomography, and muscle quality were assessed at baseline and after 10 wk of training. The IGF1 repeat promoter polymorphism and three single-nucleotide polymorphisms were genotyped. For the promoter polymorphism, subjects were grouped as homozygous for the 192 allele, heterozygous, or noncarriers of the 192 allele. After 10 wk of training, 1-repetition maximum, muscle volume, and muscle quality increased significantly for all groups combined (P < 0.001). However, carriers of the 192 allele gained significantly more strength with ST than noncarriers of the 192 allele (P = 0.02). There was also a nonsignificant trend for a greater increase in muscle volume in 192 carriers than noncarriers (P = 0.08). No significant associations were observed for the other polymorphisms studied. Thus these data suggest that the IGF1 promoter polymorphism may influence the strength response to ST. Larger sample sizes should be used in future studies to verify these results. PMID- 15894538 TI - Decreased intestinal polyp multiplicity is related to exercise mode and gender in ApcMin/+ mice. AB - Moderate-intensity treadmill running can alter male Apc(Min/+) mouse polyp formation. This purpose of this study was to examine whether exercise mode differentially affects Apc(Min/+) mouse intestinal polyp development in male and female mice. Male and female Apc(Min/+) mice were randomly assigned to control, treadmill (18 m/min; 60 min/day; 6 days/wk), or voluntary wheel running (24-h access) groups. Nine weeks of training decreased total intestinal polyps by 29% in male treadmill runners (66 +/- 9; P = 0.038) compared with male controls (93 +/- 7). The number of large polyps (>/=1-mm diameter) were also reduced by 38% in male treadmill runners (49 +/- 6; P = 0.005) compared with male controls (79 +/- 6). Treadmill running in female Apc(Min/+) mice and wheel running in both genders did not affect polyp number or size. Spleen weight decreased in male treadmill runners (91 +/- 9 mg; P = 0.011) and wheel runners (75 +/- 6 mg; P = 0.004) compared with controls (141 +/- 13 mg). Plasma IL-6 was reduced by 96% in male treadmill runners (1.2 +/- 0.6 pg/ml) and 78% in male wheel runners (6.6 +/- 3.3 pg/ml) compared with control mice (27.9 +/- 2.8 pg/ml; P < 0.05). Female mice responded similarly with an 86% decrease in plasma IL-6 with treadmill running (3.2 +/- 1.2 pg/ml) and 90% decrease with wheel running (2.9 +/- 2.0 pg/ml) compared with control mice (21.1 +/- 5.3 pg/ml; P < 0.05). The crypt depth-to villus height ratio in the intestine, an indirect marker of intestinal inflammation, decreased by 21 (P = 0.024) and 24% (P = 0.029), respectively, in male and female treadmill runners but not wheel runners. Physical activity induced attenuation of intestinal polyp number and size is dependent on exercise mode and differs between genders. The modulation of systemic and intestinal inflammation may also depend on exercise mode. PMID- 15894540 TI - Mechanical, biochemical, and extracellular matrix effects on vascular smooth muscle cell phenotype. AB - The vascular smooth muscle cell (VSMC) is surrounded by a complex extracellular matrix that provides and modulates a variety of biochemical and mechanical cues that guide cell function. Conventional two-dimensional monolayer culture systems recreate only a portion of the cellular environment, and therefore there is increasing interest in developing more physiologically relevant three-dimensional culture systems. This review brings together recent studies on how mechanical, biochemical, and extracellular matrix stimulation can be applied to study VSMC function and how the combination of these factors leads to changes in phenotype. Particular emphasis is placed on in vitro experimental studies in which multiple stimuli are combined, especially in three-dimensional culture systems and in vascular tissue engineering applications. These studies have provided new insight into how VSMC phenotype is controlled, and they have underscored the interdependence of biochemical and mechanical signaling. Future improvements in creating more complex in vitro culture environments will lead to a better understanding of VSMC biology, new treatments for vascular disease, as well as improved blood vessel substitutes. PMID- 15894541 TI - Yeast mRNA Poly(A) tail length control can be reconstituted in vitro in the absence of Pab1p-dependent Poly(A) nuclease activity. AB - Regulation of poly(A) tail length during mRNA 3'-end formation requires a specific poly(A)-binding protein in addition to the cleavage/polyadenylation machinery. The mechanism that controls polyadenylation in mammals is well understood and involves the nuclear poly(A)-binding protein PABPN1. In contrast, poly(A) tail length regulation is poorly understood in yeast. Previous studies have suggested that the major cytoplasmic poly(A)-binding protein Pab1p acts as a length control factor in conjunction with the Pab1p-dependent poly(A) nuclease PAN, to regulate poly(A) tail length in an mRNA specific manner. In contrast, we recently showed that Nab2p regulates polyadenylation during de novo synthesis, and its nuclear location is more consistent with a role in 3'-end processing than that of cytoplasmic Pab1p. Here, we investigate whether PAN activity is required for de novo poly(A) tail synthesis. Components required for mRNA 3'-end formation were purified from wild-type and pan mutant cells. In both situations, 3'-end formation could be reconstituted whether Nab2p or Pab1p was used as the poly(A) tail length control factor. However, polyadenylation was more efficient and physiologically more relevant in the presence of Nab2p as opposed to Pab1p. Moreover, cell immunofluorescence studies confirmed that PAN subunits are localized in the cytoplasm which suggests that cytoplasmic Pab1p and PAN may act at a later stage in mRNA metabolism. Based on these findings, we propose that Nab2p is necessary and sufficient to regulate poly(A) tail length during de novo synthesis in yeast. PMID- 15894542 TI - Disabled-2 (Dab2) mediates transforming growth factor beta (TGFbeta)-stimulated fibronectin synthesis through TGFbeta-activated kinase 1 and activation of the JNK pathway. AB - The multifunctional cytokine transforming growth factor beta (TGFbeta) exerts many of its effects through its regulation of extracellular matrix components, including fibronectin (FN). Although expression of both TGFbeta and FN are essential for embryonic development and wound healing in the adult, overexpression leads to excessive deposition of extracellular matrix observed in many fibroproliferative disorders. We previously have demonstrated that TGFbeta stimulated FN induction requires activation of the c-Jun N-terminal kinase (JNK) pathway; however, the signaling molecules that link the TGFbeta receptors to the JNK pathway remain unknown. We show here that the cytosolic adaptor protein disabled-2 (Dab2) directly stimulates JNK activity, whereas stable small interfering RNA-mediated ablation of Dab2 in NIH3T3 mouse fibroblasts and A10 rat aortic smooth muscle cells demonstrates that its expression is required for TGFbeta-mediated FN induction. We demonstrate that TGFbeta treatment stimulates the association of Dab2 with the mitogen-activated protein kinase kinase kinase, TAK1. Attenuation of cellular TAK1 levels by transient double-stranded RNA oligonucleotide transfection as well as overexpression of kinase-deficient TAK1 leads to abrogation of TGFbeta-stimulated FN induction. Furthermore, cell migration, another JNK-dependent response, is attenuated in NIH3T3-siDab2 expressing clones. We, therefore, delineate a signaling pathway proceeding from the TGFbeta receptors to Dab2 and TAK1, leading to TGFbeta-stimulated JNK activation, FN expression, and cell migration. PMID- 15894543 TI - The Jak1 SH2 domain does not fulfill a classical SH2 function in Jak/STAT signaling but plays a structural role for receptor interaction and up-regulation of receptor surface expression. AB - The presence of a Src homology 2 (SH2) domain sequence similarity in the sequence of Janus kinases (Jaks) has been discussed since the first descriptions of these enzymes. We performed an in depth study to determine the function of the Jak1 SH2 domain. We investigated the functionality of the Jak1 SH2 domain by stably reconstituting Jak1-defective human fibrosarcoma cells U4C with endogenous amounts of Jak1 in which the crucial arginine residue Arg466 within the SH2 domain has been replaced by lysine. This mutant still binds to the receptor subunits gp130 and OSMR. Moreover, the SH2 R466K mutation does not affect the subcellular distribution of Jak1 as assessed by cell fractionation and confocal microscopy of cells expressing endogenous levels of non-tagged or a yellow fluorescent protein (YFP)-tagged Jak1-R466K, respectively. Likewise, the signaling capacity of Jak1 was not affected by this point mutation. However, we found that the SH2 domain is structurally important for cytokine receptor binding and surface expression of the OSMR. PMID- 15894545 TI - Reconstruction of the conserved beta-bulge in mammalian defensins using D-amino acids. AB - Defensins are cationic antimicrobial mini-proteins that play important roles in the innate immune defense against microbial infection. Six invariant Cys residues in each defensin form three structurally indispensable intramolecular disulfide bridges. The only other residue invariant in all known mammalian defensins is a Gly. Structural studies indicate that the invariant Gly residue is located in an atypical, classic-type beta-bulge with the backbone torsion angles (Phi, Psi) disallowed for L-amino acids but permissible for D-enantiomers. We replaced the invariant Gly17 residue in human neutrophil alpha-defensin 2 (HNP2) by L-Ala or one of the D-amino acids Ala, Glu, Phe, Arg, Thr, Val, or Tyr. Although L-Ala17 HNP2 could not be folded, resulting in massive aggregation, all of the D-amino acid-substituted analogs folded with high efficiency. The high resolution x-ray crystal structures of dimeric D-Ala17-HNP2 were determined in three different crystal forms, showing a well preserved beta-bulge identical to those found in other defensins. The seven D-analogs of HNP2 exhibited highly variable bactericidal activity against Gram-positive and Gram-negative test strains, consistent with the premise that interplay between charge and hydrophobicity dictates how amphiphilic defensins kill. Further, the bactericidal activity of these d-amino acid analogs of HNP2 correlated well with their ability to induce leakage from large unilamellar vesicles, supporting membrane permeabilization as the lethal event in microbial killing by HNP2. Our findings identify a conformational prerequisite in the beta-bulge of defensins essential for correct folding and native structure, thereby explaining the molecular basis of the Gly Xaa-Cys motif conserved in all mammalian defensins. PMID- 15894546 TI - Cost-effectiveness analysis of breast cancer adjuvant treatment: FEC 50 versus FEC 100 (FASG05 study). AB - BACKGROUND: The aim of the study was to assess the incremental cost-effectiveness ratio (ICER) of the FEC 100 compared with the FEC 50 in the FASG05 trial. MATERIALS AND METHODS: A cost-effectiveness analysis was performed using a multi state Markov process model. Relevant clinical data introduced into the model were obtained from 10-year follow-up of the clinical trial FASG05. Survival curves for each health state were assessed by survival parametric model. The model allowed assessments from the start of adjuvant chemotherapy until death. The costs of adjuvant treatment and follow-up were estimated. The costs of recurrence were evaluated from the medical records of 146 patients. A prospective survey was performed on a cohort of 87 patients to quantify the resources external to the hospital (including cost of transportation). The inpatient costs were evaluated using the French diagnosis-related groups. The ambulatory costs were assessed using the French nomenclature. Costs were expressed in 2002 Euro (), according to the French societal perspective. The ICER assessed the cost of one additional life year saved. A discount rate of 5% per year was used for cost, and alternatively 0%, 3% and 5% for effectiveness. We validated the results with a probabilistic sensitivity analysis incorporating parametric and non-parametric bootstraps, and with the acceptability curves. RESULTS: The mean total discounting cost of adjuvant treatments was 11 465 for FEC 50 and 13 815 for FEC 100; the mean total discounting cost of recurrences was 14 636 and 13 503, respectively. According to the discount rate of effectiveness, the life expectancy was 16.5, 11.4 and 9.3 years for FEC 50 and 18.4, 12.5 and 10.2 years for FEC 100. The ICER (cost per life year saved) were 642, 1084 and 1460, respectively. The probability according to which FEC 50 is strictly dominated by FEC 100 was 0.15. CONCLUSION: The clinical benefit of FEC 100 generates a negligible cost increase when compared with FEC 50. PMID- 15894547 TI - Treatment of advanced breast cancer: the good, the bad and the ugly. PMID- 15894548 TI - A phase I-II study of weekly oxaliplatin, 5-fluorouracil continuous infusion and preoperative radiotherapy in locally advanced rectal cancer. AB - BACKGROUND: Oxaliplatin (OXA) significantly enhanced the antitumour activity of 5 fluorouracil (FUra) in patients with advanced colorectal cancer and displayed radiosensitising properties in preclinical studies. This study was thus performed to test the feasibility, identify the recommended doses (RDs) and explore preliminarily the clinical activity of weekly OXA and infused FUra combined with preoperative pelvic radiotherapy. PATIENTS AND METHODS: Forty-six patients with recurrent or locally advanced (cT3-4 and/or N+) adenocarcinomas of the mid-low rectum were treated with escalating doses of OXA (25, 35, 45, 60 mg/m2, weekly for 6 weeks) and FUra (200-225 mg/m2/day, 6-week infusion) concurrent to preoperative pelvic radiotherapy (50.4 Gy/28 fractions). The RDs for the phase II part of the study were immediately below the level resulting in dose-limiting toxicities in more than one third of the patients, or corresponded to the last planned dose level. RESULTS: In the escalation phase, dose-limiting toxicities only occurred in one patient at the fourth level and one of six patients treated at the last planned dose level (grade III diarrhoea). OXA 60 mg/m2 and FUra 225 mg/m2/day are therefore the RDs for the regimen. Among 25 patients globally treated at these doses (phase II part), the incidence of grade III diarrhoea was 16% with no grade IV toxicity. Neurotoxicity did not exceed grade II (12%). All patients completed radiotherapy and were operated on as scheduled. Twenty-one of 25 patients had the tumour down-staged after chemoradiation with seven (28%) pathological complete responses and 12 (48%) residual tumours limited to ypT1 2N0. CONCLUSIONS: Weekly OXA, at doses potentially active systemically, can be combined with full-dose, infused FUra and radiotherapy. Given the low toxicity and promising activity, this regimen is being compared to standard FUra-based pelvic chemoradiation in a randomised study. PMID- 15894549 TI - Carbohydrate concentrations in crown fractions from winter oat during hardening at sub-zero temperatures. AB - BACKGROUND AND AIMS: Contradictory results in correlation studies of plant carbohydrates with freezing tolerance may be because whole crown tissue is analysed for carbohydrates while differences exist in the survival of specific tissue within the crown. The aim of this study was to see if carbohydrate changes in tissue within oat crowns during second phase hardening (sub-zero hardening) are tissue specific. METHODS: The lower portion of oat (Avena sativa) crowns was exposed to mild grinding in a blender and the remaining crown meristem complex, consisting of tough root-like vessels, was ground in a device developed specifically for grinding cereal crown tissue. Carbohydrates were extracted by water and measured by HPLC. Carbohydrate concentrations were compared in the two regions of the crown before and after hardening at sub-zero temperatures. KEY RESULTS: Fructan of all size classes except DP>6 decreased during sub-zero hardening in both stems (base of leaf sheath) and crown meristem complex. Total simple sugar increase, including sucrose, was significantly higher in the crown meristem complex than in the stem. CONCLUSIONS: Results support the hypothesis that carbohydrate change in mildly frozen plants is tissue specific within crowns and underscore the need to evaluate specific tissue within the crown when correlating the biochemistry of plants with freezing tolerance. PMID- 15894550 TI - Phytochromes and shade-avoidance responses in plants. AB - BACKGROUND AND AIMS: The ability to detect and respond to the impending threat of shade can confer significant selective advantage to plants growing in natural communities. This Botanical Briefing highlights (a) the regulation of shade avoidance responses by endogenous and exogenous factors and (b) current understanding of the molecular components involved in red to far-red ratio signal transduction. SCOPE: The Briefing covers: (a) the shade-avoidance syndrome in higher plants; (b) the adaptive significance of shade avoidance in natural light environments; (c) phytochrome regulation of shade-avoidance responses; (d) the role of blue light signals in shade avoidance; (e) gating of rapid shade avoidance responses by the circadian clock; (f) potential signalling components and future perspectives. PMID- 15894552 TI - Newly diagnosed HIV infections: review in UK and Ireland. PMID- 15894551 TI - Cellulose biosynthesis: current views and evolving concepts. AB - AIMS: To outline the current state of knowledge and discuss the evolution of various viewpoints put forth to explain the mechanism of cellulose biosynthesis. * SCOPE: Understanding the mechanism of cellulose biosynthesis is one of the major challenges in plant biology. The simplicity in the chemical structure of cellulose belies the complexities that are associated with the synthesis and assembly of this polysaccharide. Assembly of cellulose microfibrils in most organisms is visualized as a multi-step process involving a number of proteins with the key protein being the cellulose synthase catalytic sub-unit. Although genes encoding this protein have been identified in almost all cellulose synthesizing organisms, it has been a challenge in general, and more specifically in vascular plants, to demonstrate cellulose synthase activity in vitro. The assembly of glucan chains into cellulose microfibrils of specific dimensions, viewed as a spontaneous process, necessitates the assembly of synthesizing sites unique to most groups of organisms. The steps of polymerization (requiring the specific arrangement and activity of the cellulose synthase catalytic sub-units) and crystallization (directed self-assembly of glucan chains) are certainly interlinked in the formation of cellulose microfibrils. Mutants affected in cellulose biosynthesis have been identified in vascular plants. Studies on these mutants and herbicide-treated plants suggest an interesting link between the steps of polymerization and crystallization during cellulose biosynthesis. * CONCLUSIONS: With the identification of a large number of genes encoding cellulose synthases and cellulose synthase-like proteins in vascular plants and the supposed role of a number of other proteins in cellulose biosynthesis, a complete understanding of this process will necessitate a wider variety of research tools and approaches than was thought to be required a few years back. PMID- 15894553 TI - Temporal trends in multiple births after in vitro fertilisation in Sweden, 1982 2001: a register study. PMID- 15894554 TI - Optimal search strategies for retrieving scientifically strong studies of treatment from Medline: analytical survey. AB - OBJECTIVE: To develop and test optimal Medline search strategies for retrieving sound clinical studies on prevention or treatment of health disorders. DESIGN: Analytical survey. DATA SOURCES: 161 clinical journals indexed in Medline for the year 2000. MAIN OUTCOME MEASURES: Sensitivity, specificity, precision, and accuracy of 4862 unique terms in 18 404 combinations. RESULTS: Only 1587 (24.2%) of 6568 articles on treatment met criteria for testing clinical interventions. Combinations of search terms reached peak sensitivities of 99.3% (95% confidence interval 98.7% to 99.8%) at a specificity of 70.4% (69.8% to 70.9%). Compared with best single terms, best multiple terms increased sensitivity for sound studies by 4.1% (absolute increase), but with substantial loss of specificity (absolute difference 23.7%) when sensitivity was maximised. When terms were combined to maximise specificity, 97.4% (97.3% to 97.6%) was achieved, about the same as that achieved by the best single term (97.6%, 97.4% to 97.7%). The strategies newly reported in this paper outperformed other validated search strategies except for two strategies that had slightly higher specificity (98.1% and 97.6% v 97.4%) but lower sensitivity (42.0% and 92.8% v 93.1%). CONCLUSION: New empirical search strategies have been validated to optimise retrieval from Medline of articles reporting high quality clinical studies on prevention or treatment of health disorders. PMID- 15894555 TI - Role of radiography in predicting progression of osteoarthritis of the hip: prospective cohort study. AB - OBJECTIVES: To investigate which variables identify people at high risk of progression of osteoarthritis of the hip. DESIGN: Population based cohort study. SETTING: Ommoord district in Rotterdam, Netherlands. PARTICIPANTS: 1904 men and women aged 55 years and older from the Rotterdam study were selected on the basis of the presence of osteoarthritic signs on radiography at baseline, as defined by a Kellgren and Lawrence score > or = grade 1. MAIN OUTCOME MEASURES: Radiological progression of osteoarthritis of the hip, defined as a decrease of joint space width (> or = 1.0 mm) at follow-up or the presence of a total hip replacement. METHODS: Potential determinants of progression of hip osteoarthritis were collected at baseline. x Ray films of the hip at baseline and follow-up (mean follow-up time 6.6 years) were evaluated. Multivariate logistic regression models were used to assess the association between potential risk factors and progression of hip osteoarthritis. RESULTS: In 13.1% (1904 subjects) of the study population (mean age 66.2 years), progression of hip osteoarthritis was evident on the radiograph. Starting with a simple model of only directly obtainable variables, the Kellgren and Lawrence score at baseline, when added to the model, was a strong predictor (odds ratio 5.8, 95% confidence interval 4.0 to 8.4), increasing to 24.3 (11.3 to 52.1) in subjects with hip pain at baseline. CONCLUSIONS: The Kellgren and Lawrence score at baseline was by far the strongest predictor for progression of hip osteoarthritis, especially in patients with existing hip pain at baseline. In patients with hip pain, a radiograph has strong additional value in identifying those at high risk of progression of hip osteoarthritis. PMID- 15894556 TI - Modeling the oscillation dynamics of activated airway smooth muscle strips. AB - When strips of activated airway smooth muscle are stretched cyclically, they exhibit force-length loops that vary substantially in both position and shape with the amplitude and frequency of the stretch. This behavior has recently been ascribed to a dynamic interaction between the imposed stretch and the number of actin-myosin interactions in the muscle. However, it is well known that the passive rheological properties of smooth muscle have a major influence on its mechanical properties. We therefore hypothesized that these rheological properties play a significant role in the force-length dynamics of activated smooth muscle. To test the plausibility of this hypothesis, we developed a model of the smooth muscle strip consisting of a force generator in series with an elastic component. Realistic steady-state force-length loops are predicted by the model when the force generator obeys a hyperbolic force-velocity relationship, the series elastic component is highly nonlinear, and both elastic stiffness and force generation are adjusted so that peak loop force equals isometric force. We conclude that the dynamic behavior of airway smooth muscle can be ascribed in large part to an interaction between connective tissue rheology and the force velocity behavior of contractile proteins. PMID- 15894557 TI - Differences in the translation efficiency and mRNA stability mediated by 5'-UTR splice variants of human SP-A1 and SP-A2 genes. AB - Surfactant protein A (SP-A) plays an important role in host defense, modulation of inflammatory processes, and surfactant-related functions of the lung. The human SP-A (hSP-A) locus consists of two functional genes, SP-A1 and SP-A2. Several hSP-A 5'-untranslated region (UTR) splice variants for each gene have been characterized and shown to be translated in vitro and in vivo. In this report, we investigated the role of hSP-A 5'-UTR splice variants on SP-A production and molecular mechanisms involved. We used in vitro transient expression of hSP-A 5'-UTR constructs containing luciferase as the reporter gene and quantitative real-time PCR to study hSP-A 5'-UTR-mediated gene expression. We found that 1) the four (A'D', ABD, AB'D', and A'CD') 5'-UTR splice variants under study enhanced gene expression, by increasing luciferase activity from 2.5- to 19.5-fold and luciferase mRNA from 4.3- to 8.8-fold compared with the control vector that lacked hSP-A 5'-UTR; 2) all four 5'-UTR splice variants studied regulated mRNA stability. The ABD variant exhibited the lowest rate of mRNA decay compared with the other three constructs (A'D', AB'D', and A'CD'). These three constructs also exhibited significantly lower rate of mRNA decay compared with the control vector; 3) based on the indexes of translational efficiency (luciferase activity/mRNA), ABD and AB'D' exhibited higher translational efficiency compared with the control vector, whereas the translational efficiency of each A'D' and A'CD' was lower than that of the control vector. These findings indicate that the hSP-A 5'-UTR splice variants play an important role in both SP A translation and mRNA stability. PMID- 15894558 TI - IL-6 induces neuroendocrine dedifferentiation and cell proliferation in non-small cell lung cancer cells. AB - Interleukin-6 (IL-6) has been identified as an important growth regulator of lung cancer cells. Elevation of serum levels of IL-6 has been found in a subpopulation of lung cancer patients, but rarely in patients with benign lung diseases. Approximately 15% of non-small cell lung cancer (NSCLC) tumors exhibit neuroendocrine (NE) properties (NSCLC-NE) and have been suggested to have the biological characteristics similar to small cell lung cancer (SCLC) with early metastasis and initial responsiveness to chemotherapy. We recently showed that IL 6 promotes cell proliferation and downregulates the expression of neuron-specific enolase (NSE, one of the major NE markers) in NSCLC-NE cells. In this study, we show that IL-6 stimulates a transient increase of tyrosine phosphorylation of STAT3 in a dose-dependent fashion. Inhibition of STAT3 signaling pathway by either AG-490 (JAK2-specific inhibitor) or overexpression of STAT3Y705F (a dominant-negative STAT3) reverses NSE expression in IL-6- treated NSCLC-NE cells. In addition, IL-6 induces phosphorylation and activation of p38 MAPK. SB-203580, a p38 MAPK-specific inhibitor, inhibits IL-6-induced p38 MAPK phosphorylating activity and suppresses IL-6-stimulated cell proliferation. Together, our results indicate that STAT3 signaling pathway is involved in IL-6-induced NE differentiation and that p38 MAPK is associated with IL-6-stimulated growth regulation in NSCLC-NE cells. These data suggest that both kinase pathways play critical roles in the pathogenesis of NSCLC-NE malignancies, providing new molecular targets for future therapeutic approaches. PMID- 15894559 TI - Role of IL-6 in neuroendocrine differentiation and chemosensitivity of non-small cell lung cancer. AB - Interleukin-6 (IL-6) has been shown to regulate both growth and neuroendocrine (NE) differentiation in some types of human cancer cells, and erbB2 may be a critical component of IL-6 signaling. Non-small cell lung cancer (NSCLC) tumors that demonstrate NE properties have been suggested to have biological characteristics similar to small cell lung cancers with initial responsiveness to chemotherapy. We investigated whether IL-6 is implicated in the cell growth, NE differentiation, and chemosensitivity of NSCLC-NE cells. NSCLC-NE cells were treated with exogenous IL-6, and a subclone of an IL-6-transfected NSCLC cell line that constitutively expressed IL-6 receptor was also generated. These cells were assessed for cell proliferation by cell counting and 3-[4,5-dimethylthiazol 2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays, chemosensitivity to cisplatin and etoposide by MTT assays, and NE differentiation by observing morphological changes and immunoblotting for neuron-specific enolase (NSE). The IL-6-treated cells and the IL-6-transfected cells showed enhanced cell proliferation and downregulated NSE expression, but little change in chemosensitivity. In the culture medium, IL-6-transfected cells grew as looser aggregates than the parental cells. IL-6 could not activate the erbB genes. In conclusion, IL-6 can induce cell proliferation and NE dedifferentiation but has little effect on chemosensitivity in IL-6 receptor-expressing NSCLC-NE cells. The status of NSE expression is unlikely to be a crucial factor for chemosensitivity in NSCLC cells. PMID- 15894560 TI - Propofol injection pain in children: a prospective randomized double-blind trial of a new propofol formulation versus propofol with added lidocaine. AB - BACKGROUND: The incidence of pain on injection of propofol remains unacceptably high in children, despite various strategies to reduce it. A new drug formulation of propofol has, in adult studies, been reported to cause less injection pain compared with other propofol solutions. The aim of the present prospective randomized double-blind clinical trial was to compare the incidence of pain-free injection following the use of this new formulation with that following the use of propofol with added lidocaine in children undergoing day case surgery. METHODS: Eighty-three children (age range 2-18 yr) were randomized to receive 3 mg kg(-1) of either Propofol-Lipuro (propofol dissolved in a mixture of medium- and long-chain triglycerides [MCT-LCT]; group pL, n=42) or Diprivan (propofol dissolved in long-chain triglycerides [LCT]) with added lidocaine (0.3 mg kg(-1)) (group pD, n=41). A specially trained nurse anaesthetist assessed the occurrence of injection pain using a four-graded pain scale. RESULTS: Significantly fewer patients had an entirely pain-free propofol injection in group pL (33.3%) than in group pD (61.0%) (P=0.016). CONCLUSIONS: A new MCT-LCT propofol formulation as a plain solution was associated with a higher incidence of injection pain than LCT propofol with added lidocaine when used for induction of anaesthesia in children. PMID- 15894561 TI - Pressure recording analytical method (PRAM) for measurement of cardiac output during various haemodynamic states. AB - BACKGROUND: Cardiac output (CO) can be measured using the pressure recording analytical method (PRAM), which is a new, less invasive technique allowing beat by-beat stroke volume monitoring from the pressure signals recorded in femoral or radial arteries. METHODS: We investigated PRAM by comparing its cardiac output (PRAM-CO) with paired measurements obtained by electromagnetic flowmetry (EM-CO) and by standard thermodilution (ThD-CO) during various haemodynamic states in a swine model. Nine pigs were monitored with a pulmonary artery catheter and a femoral artery catheter at baseline, in a hyperdynamic state produced by administration of dobutamine and in a hypodynamic state induced by progressive exsanguination. Bland-Altman analysis was used. RESULTS: One hundred and eight paired cardiac output values over a range of EM-CO of 1.8-10.4 litre min(-1) resulted. We found close agreement between the techniques. Mean bias between EM CO and PRAM-CO was -0.03 litre min(-1) (precision 0.58 litre min(-1)). The 95% limits of agreement were -0.61 to +0.55 litre min(-1). Similar results between ThD-CO and PRAM-CO were found. CONCLUSIONS: In a porcine model we have demonstrated accuracy of PRAM during various haemodynamic states. PRAM is a reliable tool to detect changes in cardiac output in pigs and has ability as a basic research tool. PMID- 15894562 TI - Dysaesthesia associated with sternotomy for heart surgery. AB - BACKGROUND: Chronic pain occurs in 40-50% patients following cardiac surgery. Dysaesthesia, either in the form of heightened or diminished skin sensation, are frequently associated with chronic neuropathic pain. Therefore, dysaesthesia in the early postoperative period may predict chronic pain. However, the character and causes of dysaesthesia in the early postoperative period are unknown. The aim of this study was to investigate the incidence, extent, and causes of dysaesthesia following cardiac surgery by sternotomy. METHODS: In a prospective cohort study, 50 patients undergoing sternotomy for cardiac surgery were admitted to the study: 38 underwent coronary artery bypass graft (CABG), nine valve surgery, and three combined surgery. Forty-eight hours postoperatively, acute pain was measured by four-point verbal scale. Manual pinprick and cotton wool brushing was used to detect the areas of dysaesthesia. RESULTS: Some form of dysaesthesia was found in 27 (54%) of the patients. Using multivariate regression analysis, the total area of dysaesthesia was positively associated with CABG surgery and the severity of postoperative pain (P<0.001). CONCLUSION: Dysaesthesia is common in the early postoperative period following cardiac surgery using a sternotomy and is associated with CABG surgery. The association with severity of pain may indicate a neuropathic element that is unrelieved by conventional opioid analgesia. PMID- 15894563 TI - The decrease of the cytoskeleton tubulin follows the decrease of the associating molecular chaperone alphaB-crystallin in unloaded soleus muscle atrophy without stretch. AB - The cytoskeletal component tubulin/microtubule commonly allows the cell to respond mechanically to the environment. The concentration of free tubulin dimer is autoregulated in the balance of free dimer and polymeric forms of microtubule (MT) protein, having an intrinsic property of "dynamic instability", and through cotranslational beta-tubulin mRNA degradation. Recently, we have demonstrated that alphaB-crystallin is a key molecule of muscle atrophy, since alphaB crystallin has a chaperone-like-activity that suppresses tubulin aggregation and protects the MT disassembly against both Ca2+ and depolymelizing alkaloid in vitro. Most of the small heat-shock proteins (sHsps), including alphaB crystallin, are expressed in skeletal muscle. However, no report to date has studied the changes of tubulin/MT during muscle adaptation. Here, we examined changes in tubulin content in rat soleus muscles after hindlimb suspension (HS) with/without passive stretch and the recovery. HS induced rapid decreases of soleus muscle mass, most Hsps (alphaB-crystallin, Hsp90, Hsp70, Hsp27, and p20) and tubulin contents in soleus muscle, while heat-shock cognate 70-kDa protein (Hsc70) did not decrease. Soleus muscle mass, most Hsps, and tubulin were maintained with passive stretch. After 5 days' recovery, the levels of tubulin and Hsps, but not Hsc70, were restored to control levels. The interactions of alphaB-crystallin and tubulin/MT were observed with immunoprecipitation with an anti-alpha-tubulin antibody and taxol-dependent MT assembly. Other sHsps were also associated with alphaB-crystallin and MT, whereas Hsp90 and Hsp70 did not co precipitate with them. These data imply an interaction and close relationship between alphaB-crystallin and tubulin/MTs in muscle tissues. The amount of mRNA of alphaB-crystallin decreased with the muscle atrophy level, whereas the gene expression level of betaI-tubulin was maintained during HS. This means a significant role of post-transcriptional regulation in tubulin/MT system in muscle adaptation, whereas alphaB-crystallin and most sHsps are regulated at the transcriptional level. Additional functional contribution of alphaB-crystallin to tubulin/MTs during myotube formation was examined using C2C12 myoblast cultured cells, the alphaB-crystallin expression of which was decreased or increased. It indicated the necessity of alphaB-crystallin during microtubule reorganization. In conclusion, tubulin/MTs were revealed to be one of the substrates of alphaB crystallin, and also serial decreases of alphaB-crystallin and tubulin/MT in early soleus muscle atrophy suggest that the chaperone effect of alphaB crystallin on the cytoskeleton, which may be also dynamically regulated in the muscle cell, is a key mechanism for muscle adaptation and protection of the atrophy and also muscle differentiation. PMID- 15894564 TI - Glucagon-like peptide-1, corticotropin-releasing hormone, and hypothalamic neuronal histamine interact in the leptin-signaling pathway to regulate feeding behavior. AB - Glucagon-like peptide-1 (GLP-1), corticotropin-releasing hormone (CRH), and hypothalamic neuronal histamine suppress food intake, a target of leptin action in the brain. This study examined the interactions of GLP-1, CRH, and histamine downstream from the leptin-signaling pathway in regulating feeding behavior. Infusion of GLP-1 into the third cerebral ventricle (i3vt) at a dose of 1 mug significantly decreased the initial 1 h cumulative food intake in rats as compared with phosphate-buffered saline (PBS) controls. The GLP-1-induced suppression of feeding was partially attenuated by intraperitoneal pretreatment with alpha-fluoromethylhistidine (FMH), a specific suicide inhibitor of histidine decarboxylase, which depletes hypothalamic neuronal histamine. Pretreatment with alpha-helical CRH (10 microg/rat, i3vt), a nonselective CRH antagonist, abolished the GLP-1-induced suppression of feeding completely. I3vt infusion of GLP-1 increased the CRH content and histamine turnover assessed using the pargyline induced accumulation of tele-methyl histamine (t-MH), a major metabolite of neuronal histamine, in the hypothalamus. The central infusion of CRH also induced the increase of histamine turnover and CRH receptor type 1 was localized on the cell body of histamine neuron. Pretreatment with exendin(9-39), a GLP-1 receptor antagonist, attenuated the leptin-induced increase in CRH content of the hypothalamus. Finally, i3vt infusion of leptin also increased histamine turnover in the hypothalamus. Pretreatment with exendin(9-39), alpha-helical CRH or both antagonists attenuated the leptin-induced responses of t-MH levels in the hypothalamus. These results suggest that CRH or hypothalamic neuronal histamine mediates the GLP-1-induced suppression of feeding behavior, that CRH mediates GLP 1 signaling to neuronal histamine and that a functional link from GLP-1 to neuronal histamine via CRH constitutes the leptin-signaling pathway regulating feeding behavior. PMID- 15894565 TI - A role for endocannabinoids in the generation of parkinsonism and levodopa induced dyskinesia in MPTP-lesioned non-human primate models of Parkinson's disease. AB - Endocannabinoids and cannabinoid CB1 receptors play a role in the control of movement by modulating GABA, glutamate, and other neurotransmitters throughout the basal ganglia. Roles for abnormalities in endocannabinoid signaling in Parkinson's disease (PD) and the major side effect of current treatments, levodopa-induced dyskinesia (LID), have been suggested by rodent studies. Here we show that signaling by endocannabinoids contributes to the pathophysiology of parkinsonism and LID in MPTP-lesioned, non-human primate models of Parkinson's disease. In MPTP-lesioned marmosets previously treated with levodopa to establish LID, attenuation of CB1 signaling by systemic administration of rimonabant (1 and 3 mg/kg) had anti-parkinsonian actions, equivalent to a 71% increase in motor activity at 3 mg/kg. Rimonabant did not elicit dyskinesia. Co-administration of levodopa (8 mg/kg) and rimonabant (1 and 3 mg/kg) resulted in significantly less dyskinesia than levodopa alone, without significantly affecting the anti parkinsonian action of levodopa. These data suggest that enhanced endocannabinoid signaling may be involved in the pathophysiology of both parkinsonism and LID. To define potential mechanisms by which such a role might be mediated, we determined the levels of the endocannabinoids anandamide and 2-arachidonyl glycerol (2-AG) throughout the basal ganglia in normal and three groups of MPTP-lesioned cynomolgus monkeys (untreated; acutely treated with L-DOPA, non-dyskinetic; long term treated, with levodopa-induced dyskinesia). In the untreated, MPTP-lesioned primate, parkinsonism was associated with increases in both 2-AG (+88%) and anandamide (+49%) in the striatum, and of 2-AG (+97%) in the substantia nigra, changes that are consistent with the previously suggested role for endocannabinoids in mechanisms attempting to compensate for loss of dopamine in untreated parkinsonism. Increased levels of anandamide (+34%) in the external globus pallidus of MPTP-lesioned animals were normalized by levodopa treatment and may contribute to the generation of parkinsonian symptoms. However, no clear alteration in endocannabinoid levels could be correlated with the expression of LID. These data highlight the potential roles played by endocannabinoids and CB1 in PD and LID and suggest the need for further research to pursue the multiple therapeutic opportunities for manipulating this system in movement disorders. PMID- 15894566 TI - Human eNOS gene delivery attenuates cold-induced elevation of blood pressure in rats. AB - We previously showed that chronic cold exposure inhibits endothelial nitric oxide synthase (eNOS) expression and decreases nitric oxide (NO) production. The aim of the present study was to evaluate the possible role of the NO system in the development of cold-induced hypertension (CIH) by testing the hypothesis that adenoviral delivery of human eNOS gene increases NO production and attenuates CIH in rats. The effect of in vivo delivery of adenovirus carrying human eNOS full length cDNA (rAdv.heNOS) on CIH was tested using four groups of Sprague-Dawley rats (6 rats/group). Blood pressure (BP) did not differ among the four groups during the control period at room temperature (24 degrees C). Two groups of rats received intravenous injection of rAdv.heNOS (1 x 10(9) plaque-forming units/rat), and the other two groups received the same dose of rAdv.LacZ to serve as controls. After gene delivery, one rAdv.heNOS-treated group and one rAdv.LacZ treated group were exposed to cold (6 degrees C) while the remaining groups were kept at 24 degrees C. We found that the BP of the rAdv.LacZ group increased significantly within 1 wk of exposure to cold and reached a peak level at week 5 (152.2 +/- 6.4 mmHg). In contrast, BP (118.7 +/- 8.4 mmHg) of the cold-exposed rAdv.heNOS group did not increase until 5 wk after exposure to cold. The rAdv.heNOS increased plasma and urine levels of NO significantly in cold-exposed rats, which indicates that eNOS gene transfer increased NO production. Notably, rAdv.heNOS decreased plasma levels of norepinephrine and plasma renin activity in cold-exposed rats, which suggests that eNOS gene transfer may decrease the activities of the sympathetic nervous system and the renin-angiotensin system. Immunohistochemical analysis showed that the transferred human eNOS was expressed in both endothelium and adventitia of mesenteric arteries. We conclude that 1) eNOS gene transfer attenuates CIH by increasing NO production and inhibiting the sympathetic nervous system and the renin-angiotensin system; and 2) the NO system appears to mediate this nongenetic, nonpharmacological, nonsurgical model of hypertension. PMID- 15894567 TI - Circadian periodicity of cerebral blood flow revealed by laser-Doppler flowmetry in awake rats: relation to blood pressure and activity. AB - Cardiovascular parameters such as arterial blood pressure (ABP) and heart rate display pronounced circadian variation. The present study was performed to detect whether there is a circadian periodicity in the regulation of cerebral perfusion. Normotensive Sprague-Dawley rats (SDR, approximately 15 wk old) and hypertensive (mREN2)27 transgenic rats (TGR, approximately 12 wk old) were instrumented in the abdominal aorta with a blood pressure sensor coupled to a telemetry system for continuous recording of ABP, heart rate, and locomotor activity. After 5-12 days, a laser-Doppler flow (LDF) probe was attached to the skull by means of a guiding device to measure changes in brain cortical blood flow (CBF). After the animals recovered from anesthesia, measurements were taken for 3-4 days. The time series were analyzed with respect to the midline estimating statistic of rhythm (i.e., mean value of a periodic event after fit to a cosine function), amplitude, and acrophase (i.e., phase angle that corresponds to the peak of a given period) of the 24-h period. The LDF signal displayed a significant circadian rhythm, with the peak occurring at around midnight in SDR and TGR, despite inverse periodicity of ABP in TGR. This finding suggests independence of LDF periodicity from ABP regulation. Furthermore, the acrophase of the LDF was consistently found before the acrophase of the activity. From the present data, it is concluded that there is a circadian periodicity in the regulation of cerebral perfusion that is independent of circadian changes in ABP and probably is also independent of locomotor activity. The presence of a circadian periodicity in CBF may have implications for the occurrence of diurnal alterations in cerebrovascular events in humans. PMID- 15894568 TI - PKC-{epsilon}-dependent survival signals in diabetic hearts. AB - Diabetes mellitus is complicated by the development of a primary cardiomyopathy, which contributes to the excess morbidity and mortality of this disorder. The protein kinase C (PKC) family of isozymes plays a key role in the cardiac phenotype expressed during postnatal development and in response to pathological stimuli. Hyperglycemia is an activating signal for cardiac PKC isozymes that modulate a myriad of cell events including cell death and survival. The epsilon isozyme of the PKC family transmits a powerful survival signal in cardiac muscle cells. Accordingly, to test the hypothesis that endogenous activation of cardiac PKC-epsilon will protect against hyperglycemic cell injury and left ventricular dysfunction, diabetes mellitus was induced using streptozotocin in genetically engineered mice with cardiac-specific expression of the PKC-epsilon translocation activator [psiepsilon-receptors for activated C kinase (psiepsilon-RACK)]. The results demonstrate a striking PKC-epsilon cardioprotective phenotype in diabetic psiepsilon-RACK (epsilon-agonist) mice that is characterized by inhibition of the hyperglycemia apoptosis signal, attenuation of hyperglycemia-mediated oxidative stress, and preservation of parameters of left ventricular pump function. Hearts of diabetic epsilon-agonist mice exhibited selective trafficking of PKC-epsilon to membrane and mitochondrial compartments, phosphorylation/inactivation of the mitochondrial Bad protein, and inhibition of cytochrome c release. We conclude that activation of endogenous PKC-epsilon in hearts of diabetic epsilon-agonist mice promotes the survival phenotype, attenuates markers of oxidative stress, and inhibits the negative inotropic properties of chronic hyperglycemia. PMID- 15894569 TI - Effects of spironolactone and eprosartan on cardiac remodeling and angiotensin converting enzyme isoforms in rats with experimental heart failure. AB - Angiotensin-converting enzyme (ACE)-2 is a newly described enzyme with antagonistic effects to those of the classical ACE (ACE-1). Both ANG II and aldosterone play an important role in the pathophysiology of congestive heart failure (CHF) and in the adverse cardiac remodeling during its development. In this study, we examined the effects of experimental CHF induced by an aortocaval fistula (ACF) and of its treatment with ANG II and aldosterone inhibitors on the relative levels of ACE-1 and ACE-2. We also compared the effects of spironolactone, an aldosterone antagonist, and eprosartan, an ANG II receptor antagonist, on heart hypertrophy and fibrosis in rats with ACF. Spironolactone (15 mg x kg(-1) x day(-1) ip, via minipump) or eprosartan (5 mg x kg(-1) x day( 1) ip, via minipump) was administered into rats with ACF for 14 and 28 days. Specific antibodies were used to determine the protein levels of myocardial ACE-1 and ACE-2. ACF increased the cardiac levels of ACE-1 and decreased those of ACE 2. Heart-to-body weight ratio significantly increased from 0.30 +/- 0.004% in sham-operated controls to 0.50 +/- 0.018% and 0.56 +/- 0.044% (P < 0.001) in rats with ACF, 2 and 4 wk after surgery, respectively, in association with increased plasma levels of aldosterone. The area occupied by collagen increased from 2.33 +/- 0.27% to 6.85 +/- 0.65% and 8.03 +/- 0.93% (P < 0.01), 2 and 4 wk after ACF, respectively. Both spironolactone and eprosartan decreased cardiac mass and collagen content and reversed the shift in ACE isoforms. ACF alters the ratio between ACE isoforms in a manner that increases local ANG II and aldosterone levels. Early treatment with both ANG II and aldosterone antagonists is effective in reducing this effect. Thus ACE isoform shift may represent an important component of the development of cardiac remodeling in response to hemodynamic overload, and its correction may contribute to the beneficial therapeutic effects of renin-angiotensin-aldosterone system inhibitors. PMID- 15894570 TI - Preserved left ventricular structure and function in mice with cardiac sympathetic hyperinnervation. AB - Cardiac-specific overexpression of nerve growth factor (NGF), a neurotrophin, leads to sympathetic hyperinnervation of heart. As a consequence, adverse functional changes that occur after chronically enhanced sympathoadrenergic stimulation of heart might develop in this model. However, NGF also facilitates synaptic transmission and norepinephrine uptake, effects that would be expected to restrain such deleterious outcomes. To test this, we examined 5- to 6-mo-old transgenic (TG) mice that overexpress NGF in heart and their wild-type (WT) littermates using echocardiography, invasive catheterization, histology, and catecholamine assays. In TG mice, hypertrophy of the right ventricle was evident (+67%), but the left ventricle was only mildly affected (+17%). Left ventricular (LV) fractional shortening and fractional area change values as indicated by echocardiography were similar between the two groups. Catheterization experiments revealed that LV +/-dP/dt values were comparable between TG and WT mice and responded similarly upon isoproterenol stimulation, which indicates lack of beta adrenergic receptor dysfunction. Although norepinephrine levels in TG LV tissue were approximately twofold those of WT tissue, TG plasma levels of the neuronal norepinephrine metabolite dihydroxyphenylglycol were fivefold those of WT plasma. A greater neuronal uptake activity was also observed in TG LV tissue. In conclusion, overexpression of NGF in heart leads to sympathetic hyperinnervation that is not associated with detrimental effects on LV performance and is likely due to concomitantly enhanced norepinephrine neuronal uptake. PMID- 15894571 TI - Functional changes in adenylyl cyclases and associated decreases in relaxation responses in mesenteric arteries from diabetic rats. AB - To assess the functional change in adenylyl cyclases (AC) associated with the diabetic state, we investigated AC-mediated relaxations and cAMP production in mesenteric arteries from rats with streptozotocin (STZ)-induced diabetes. The relaxations induced by the water-soluble forskolin (FSK) analog NKH477, which is a putative AC5 activator, but not by the beta-adrenoceptor agonist isoproterenol (Iso) and the AC activator FSK, were reduced in intact diabetic mesenteric artery. In diabetic rats, however, Iso-, FSK-, and NKH477-induced relaxations were attenuated in the presence of inhibitors of nitric oxide synthase and cyclooxygenase. To exclude the influence of phosphodiesterase (PDE), we also examined the relaxations induced by several AC activators in the presence of 3 isobutyl-1-methylxanthine (IBMX; a PDE inhibitor). Under these conditions, the relaxation induced by Iso was greatly impaired in STZ-diabetic rats. This Iso induced relaxation was significantly attenuated by pretreatment with SQ-22536, an AC inhibitor, in mesenteric rings from age-matched controls but not in those from STZ-diabetic rats. Under the same conditions, the relaxations induced by FSK or NKH477 were impaired in STZ-diabetic rats. Neither FSK- nor A-23187 (a Ca2+ ionophore)-induced cAMP production was significantly different between diabetics and controls. However, cAMP production induced by Iso or NKH477 was significantly impaired in diabetic mesenteric arteries. Expression of mRNAs and proteins for AC5/6 was lower in diabetic mesenteric arteries than in controls. These results suggest that AC-mediated relaxation is impaired in the STZ-diabetic rat mesenteric artery, perhaps reflecting a reduction in AC5/6 activity. PMID- 15894572 TI - Cellular energy status modulates translational control mechanisms in ischemic reperfused rat hearts. AB - Mechanisms regulating ischemia and reperfusion (I/R)-induced changes in mRNA translation in the heart are poorly defined, as are the factors that initiate these changes. Because cellular energy status affects mRNA translation under physiological conditions, it is plausible that I/R-induced changes in translation may in part be a result of altered cellular energy status. Therefore, the purpose of the studies described herein was to compare the effects of I/R with those of altered energy substrate availability on biomarkers of mRNA translation in the heart. Isolated adult rat hearts were perfused with glucose or a combination of glucose plus palmitate, and effects of I/R on various biomarkers of translation were subsequently analyzed. When compared with hearts perfused with glucose plus palmitate, hearts perfused with glucose alone exhibited increased phosphorylation of eukaryotic elongation factor (eEF)2, the alpha-subunit of eukaryotic initiation factor (eIF)2, and AMP-activated protein kinase (AMPK), and these hearts also exhibited enhanced association of eIF4E with eIF4E binding protein (4E-BP)1. Regardless of the energy substrate composition of the buffer, phosphorylation of eEF2 and AMPK was greater than control values after ischemia. Phosphorylation of eIF2alpha and eIF4E and the association of eIF4E with 4E-BP1 were also greater than control values after ischemia but only in hearts perfused with glucose plus palmitate. Reperfusion reversed the ischemia-induced increase in eEF2 phosphorylation in hearts perfused with glucose and reversed ischemia induced changes in eIF4E, eEF2, and AMPK phosphorylation in hearts perfused with glucose plus palmitate. Because many ischemia-induced changes in mRNA translation are mimicked by the removal of a metabolic substrate under normal perfusion conditions, the results suggest that cellular energy status represents an important modulator of I/R-induced changes in mRNA translation. PMID- 15894573 TI - Effects of hyperthyroidism on delayed rectifier K+ currents in left and right murine atria. AB - Hyperthyroidism has been associated with atrial fibrillation (AF); however, hyperthyroidism-induced ion channel changes that may predispose to AF have not been fully elucidated. To understand the electrophysiological changes that occur in left and right atria with hyperthyroidism, the patch-clamp technique was used to compare action potential duration (APD) and whole cell currents in myocytes from left and right atria from both control and hyperthyroid mice. Additionally, RNase protection assays and immunoblotting were performed to evaluate the mRNA and protein expression levels of K(+) channel alpha-subunits in left and right atria. The results showed that 1) in control mice, the APD was shorter and the ultra-rapid delayed rectifier K(+) conductance (I(Kur)) and the sustained delayed rectifier K(+) conductance (I(ss)) were larger in the left than in the right atrium; also, mRNA and protein expression levels of Kv1.5 and Kv2.1 were higher in the left atrium; 2) in hyperthyroid mice, the APD was shortened and I(Kur) and I(ss) were increased in both left and right atrial myocytes, and the protein expression levels of Kv1.5 and Kv2.1 were increased significantly in both atria; and 3) the influence of hyperthyroidism on APD and delayed rectifier K(+) currents was more prominent in right than in left atrium, which minimized the interatrial APD difference. In conclusion, hyperthyroidism resulted in more significant APD shortening and greater delayed rectifier K(+) current increases in the right vs. the left atrium, which can contribute to the propensity for atrial arrhythmia in hyperthyroid heart. PMID- 15894574 TI - Acute systemic hypoxia elevates venous but not interstitial potassium of dog skeletal muscle. AB - Potassium release through ATP-sensitive potassium (K(ATP)) channels contributes to hypoxic vasodilation in the skeletal muscle vascular bed: It is uncertain whether K(ATP) channels on muscle cells contribute to the process. Potassium from muscle cells must cross the interstitial space to reach the vascular tissues, whereas that from vascular endothelium would have a higher concentration in venous blood than in interstitial fluid. We determined the effect of systemic hypoxia on arterial, venous, and interstitial potassium in the constant-flow perfused gracilis muscles of anesthetized dogs. Hypoxia reduced arterial Po(2) from 138 to 25 and Pco(2) from 28 to 26 mmHg. Arterial pH and potassium were well correlated (r(2) = 0.9): Both increased in early hypoxia and decreased during the postcontrol. In denervated muscles, perfusion pressure decreased from 95 to 76 mmHg by the end of the hypoxic period; neither venous nor interstitial potassium was elevated. In innervated muscles, perfusion pressure increased from 110 to 172 mmHg by the 11th min of hypoxia and then decreased to 146 mmHg by the end of the hypoxic period; venous potassium increased from 5.0 to 5.3 mM, but interstitial potassium remained unchanged. Glibenclamide abolished both the increase in venous potassium and the hypoxic vasodilation in the innervated muscle. Thus skeletal muscle cells were unlikely to have contributed to the release of potassium, which was suggested to originate from vascular endothelium. The sympathetic nerve supply may play a direct or indirect role in the opening of K(ATP) channels under hypoxic conditions. PMID- 15894575 TI - Effect of mechanical stretch on HIF-1{alpha} and MMP-2 expression in capillaries isolated from overloaded skeletal muscles: laser capture microdissection study. AB - Under physiological nonhypoxic conditions, angiogenesis can be driven by mechanical forces. However, because of the limitations of the specific gene expression analysis of microvessels from in vivo experiments, the mechanisms regulating the coordinated expression of angiogenic factors implicated in the process remain intangible. In this study, the technique of laser capture microdissection (LCM) was adapted for the study of angiogenesis in skeletal muscles. With a combination of LCM and real-time quantitative PCR it was demonstrated that capillary endothelial cells produce matrix metalloproteinase (MMP)-2 and that mechanical stretch of capillaries within muscle tissue markedly increases MMP-2 mRNA (2.5-fold increase vs. control; P < 0.05). In addition, we showed that transcription factor hypoxia-inducible factor (HIF)-1alpha expression was 13.5-fold higher in capillaries subjected to stretch compared with controls (P < 0.05). These findings demonstrate the feasibility of this approach to study angiogenic gene regulation and provide novel evidence of HIF-1alpha induction in stretched capillary endothelial cells. PMID- 15894576 TI - Role of nitric oxide scavenging in vascular response to cell-free hemoglobin transfusion. AB - Modified Hb solutions have been developed as O(2) carrier transfusion fluids, but of concern is the possibility that increased scavenging of nitric oxide (NO) within the plasma will alter vascular reactivity even if the Hb does not readily extravasate. The effect of decreasing hematocrit from approximately 30% to 18% by an exchange transfusion of a 6% sebacyl cross-linked tetrameric Hb solution on the diameter of pial arterioles possessing tight endothelial junctions was examined through a cranial window in anesthetized cats with and without a NO synthase (NOS) inhibitor. Superfusion of a NOS inhibitor decreased diameter, and subsequent Hb transfusion produced additional constriction that was not different from Hb transfusion alone but was different from the dilation observed by exchange transfusion of an albumin solution after NOS inhibition. In contrast, abluminal application of the cross-linked Hb produced constriction that was attenuated by the NOS inhibitor. Neither abluminal nor intraluminal cross-linked Hb interfered with pial arteriolar dilation to cromakalim, an activator of ATP sensitive potassium channels. Pial vascular reactivity to hypocapnia and hypercapnia was unaffected by Hb transfusion. Microsphere-determined regional blood flow indicated selective decreases in perfusion after Hb transfusion in the kidney, small intestine, and neurohypophysis, which does not have tight endothelial junctions. Administration of a NOS inhibitor to reduce the basal level of NO available for scavenging before Hb transfusion prevented further decreases in blood flow to these regions compared with NOS inhibition alone. In contrast, blood flow to skeletal and left ventricular muscle increased, and cerebral blood flow was unchanged after Hb transfusion. This cross-linked Hb tetramer is known to appear in renal lymph but not in urine. We conclude that cell-free tetrameric Hb does not scavenge sufficient NO in the plasma space to significantly affect baseline tone in vascular beds with tight endothelial junctions but does produce substantial constriction in beds with porous endothelium. The data support increasing the molecular size of Hb by polymerization or conjugation to limit extravasation in all vascular beds to preserve normal vascular reactivity. PMID- 15894577 TI - Leptin resistance extends to the coronary vasculature in prediabetic dogs and provides a protective adaptation against endothelial dysfunction. AB - Hyperleptinemia, associated with prediabetes, is an independent risk factor for coronary artery disease and a mediator of coronary endothelial dysfunction. We previously demonstrated that acutely raising the leptin concentration to levels comparable with those observed in human obesity significantly attenuates coronary dilation/relaxation to acetylcholine (ACh) both in vivo in anesthetized dogs and in vitro in isolated canine coronary rings. Accordingly, the purpose of this investigation was to extend these studies to a model of prediabetes with chronic hyperleptinemia. In the present investigation, experiments were conducted on control and high-fat-fed dogs. High-fat feeding caused a significant increase (131%) in plasma leptin concentration. Furthermore, in high-fat-fed dogs, exogenous leptin did not significantly alter vascular responses to ACh in vivo or in vitro. Coronary vasodilator responses to ACh (0.3-30.0 microg/min) and sodium nitroprusside (1.0-100.0 microg/min) were not significantly different from those observed in control dogs. Also, high-fat feeding did not induce a switch to an endothelium-derived hyperpolarizing factor as a major mediator of muscarinic coronary vasodilation, because dilation to ACh was abolished by combined pretreatment with N(omega)-nitro-l-arginine methyl ester (150 microg/min ic) and indomethacin (10 mg/kg iv). Quantitative, real-time PCR revealed no significant difference in coronary artery leptin receptor gene expression between control and high-fat-fed dogs. In conclusion, high-fat feeding induces resistance to the coronary vascular effects of leptin, and this represents an early protective adaptation against endothelial dysfunction. The resistance is not due to altered endothelium-dependent or -independent coronary dilation, increased endothelium derived hyperpolarizing factor, or changes in coronary leptin receptor mRNA levels. PMID- 15894578 TI - Effect of serum triiodothyronine on regulation of cardiac gene expression: role of histone acetylation. AB - Thyroid hormone regulates the transcription of several important cardiac genes. Although the thyroid gland produces predominantly thyroxine (T(4)), it is triiodothyronine (T(3)) that is transported across the sarcolemma and binds to nuclear thyroid hormone receptor proteins; yet various studies suggest that serum T(3) levels do not accurately reflect cellular T(3) action. To address this question, we studied the dose-response relationship of T(3) administered by constant infusion in hypothyroid animals with the simultaneous in vivo transcription rate of the cardiac-specific alpha-myosin heavy chain (MHC) gene, measured by quantitating alpha-MHC heteronuclear (hn)RNA content. Constant infusion of 4 mug T(3) x kg body wt(-1) x day(-1) for 3 days normalized serum T(3) and restored transcription to euthyroid levels; in contrast, daily injections of the same dose increased alpha-MHC transcription by only 55% of that obtained by infusion. Although infusion of T(3) at 1.25 microg T(3) x kg body wt( 1) x day(-1) was not sufficient to restore serum T(3) to normal, it was capable of restoring transcription to normal at 3 days, but when administered for 12 days, transcription of alpha-MHC was found to be 50% of euthyroid levels, demonstrating a decreased sensitivity to T(3) over time. Treatment with trichostatin A (TSA) to inhibit histone deacetylation increased levels of total nuclear acetylated histone H4 by almost 50% but was without effect on the real time PCR measures of alpha-MHC hnRNA. TSA administered together with T(3) (10 mug T(3)/kg body wt) significantly increased transcription of alpha-MHC after 30 h, thus demonstrating a potential role for histones as cofactors in the T(3) regulation of cardiac alpha-MHC transcription. PMID- 15894579 TI - Focal gap junction uncoupling and spontaneous ventricular ectopy. AB - Genetic studies in the mouse have demonstrated that conditional cardiac restricted loss of connexin43 (Cx43), the major ventricular gap junction protein, is highly arrhythmogenic. However, whether more focal gap junction remodeling, as is commonly seen in acquired cardiomyopathies, influences the propensity for arrhythmogenesis is not known. We examined electrophysiological properties and the frequency of spontaneous and inducible arrhythmias in genetically engineered chimeric mice derived from injection of Cx43-deficient embryonic stem cells into wild-type recipient blastocysts. Chimeric mice had numerous well-circumscribed microscopic Cx43-negative foci in their hearts, comprising approximately 15% of the total surface area as determined by immunohistochemical analysis. Systolic function in the chimeric mice was significantly depressed as measured echocardiographically (19.0% decline in fractional shortening compared with controls, P < 0.05) and by invasive hemodynamics (17.6% reduction in change of pressure over time, P < 0.01). Chimeras had significantly more spontaneous arrhythmic events than controls (P < 0.01), including frequent runs of nonsustained ventricular tachycardia in some of the chimeric mice. However, in contrast to mice with conditional cardiac-resricted loss of Cx43 in the heart, no sustained ventricular tachyarrhythmias were observed. We conclude that focal areas of uncoupling in the myocardium increase the likelihood of arrhythmic triggers, but more widespread uncoupling is required to support sustained arrhythmias. PMID- 15894580 TI - Functional alterations in cerebrovascular K(+) and Ca(2+) channels are comparable between simulated microgravity rat and SHR. AB - Exposure to microgravity leads to a sustained elevation in transmural pressure across the cerebral vasculature due to removal of hydrostatic pressure gradients. We hypothesized that ion channel remodeling in cerebral vascular smooth muscle cells (VSMCs) similar to that associated with hypertension may occur and play a role in upward autoregulation of cerebral vessels during microgravity. Sprague Dawley rats were subjected to 4-wk tail suspension (Sus) to simulate the cardiovascular effect of microgravity. Large-conductance Ca(2+)-activated K(+) (BK(Ca)), voltage-gated K(+) (K(V)), and L-type voltage-dependent Ca(2+) (Ca(L)) currents of Sus and control (Con) rat cerebral VSMCs were investigated with a whole cell voltage-clamp technique. Under the same experimental conditions, K(V), BK(Ca), and Ca(L) currents of cerebral VSMCs from adult spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) were also investigated. K(V) current density decreased in Sus rats vs. Con rats [1.07 +/- 0.14 (n = 22) vs. 1.31 +/- 0.28 (n = 16) pA/pF at +20 mV (P < 0.05)] and BK(Ca) and Ca(L) current densities increased [BK(Ca): 1.70 +/- 0.37 (n = 23) vs. 0.88 +/- 0.22 (n = 19) pA/pF at +20 mV (P < 0.05); Ca(L): -2.17 +/- 0.21 (n = 35) vs. -1.31 +/- 0.10 (n = 26) pA/pF at +10 mV (P < 0.05)]. Similar changes were also observed in SHR vs. WKY cerebral VSMCs: K(V) current density decreased [1.03 +/- 0.33 (n = 9) vs. 1.62 +/- 0.64 (n = 9) pA/pF at +20 mV (P < 0.05)] and BK(Ca) and Ca(L) current densities increased [BK(Ca): 2.54 +/- 0.47 (n = 11) vs. 1.12 +/- 0.33 (n = 12) pA/pF at +20 mV (P < 0.05); Ca(L): -3.99 +/- 0.53 (n = 12) vs. -2.28 +/- 0.20 (n = 10) pA/pF at +20 mV (P < 0.05)]. These findings support our hypothesis, and their impact on space cardiovascular research is discussed. PMID- 15894581 TI - Use of translucent indium tin oxide to measure stimulatory effects of a passive conductor during field stimulation of rabbit hearts. AB - Biomathematical models and experiments have indicated that passive extracellular conductors influence field stimulation. Because metallic conductors prevent optical mapping under the conductor, we have evaluated a passive translucent indium tin oxide (ITO) thin-film conductor to allow mapping of transmembrane potential (V(m)) and stimulatory current under the conductor. A 1-cm ITO disk was patterned photolithographically and positioned between 0.3-cm(2) mesh shock electrodes on the ventricular epicardium of isolated perfused rabbit hearts stained with 4-{2-[6-(dibutylamino)-2-naphthylenal]ethenyl}-1-(3-sulfopropyl)-, hydroxide, inner salt (di-4-ANEPPS). For a 1-A, 10-ms shock during the action potential plateau, optical maps from fluorescence collected using emission ratiometry (excitation at 488 nm and emissions at 510-570 and >590 nm) indicated that the disk altered V(m) by as much as the height of an action potential. DeltaV(m) became more positive near the edge of the disk, where the ITO conductance gradient was parallel to applied current, and more negative near the opposite edge, where the gradient was not parallel to current. For diastolic shocks, the disk expedited membrane excitation at the sites of positive DeltaV(m) in the heart and in a cardiac model with realistic ITO disk surface and interfacial conductances. Optical maps of ITO transmittance and the model indicated that the disk introduced anodal and cathodal stimulatory current at opposite edges of the disk. Thus ITO allows study of the stimulatory effects of a passive conductor in an electric field. PMID- 15894582 TI - Escherichia coli K1 inhibits proinflammatory cytokine induction in monocytes by preventing NF-kappaB activation. AB - Phagocytes are well-known effectors of the innate immune system to produce proinflammatory cytokines and chemokines such as tumor necrosis factor alpha (TNF alpha), interleukin (IL)-1beta, and IL-8 during infections. Here, we show that infection of monocytes with wild-type Escherichia coli K1, which causes meningitis in neonates, suppresses the production of cytokines and chemokines (TNF-alpha, regulated on activation, normal T expressed and secreted, macrophage inflammatory protein-1beta, IL-1beta, and IL-8). In contrast, infection of monocytes with a mutant E. coli, which lacks outer membrane protein A (OmpA- E. coli) resulted in robust production of cytokines and chemokines. Wild-type E. coli K1 (OmpA+ E. coli) prevented the phosphorylation and its degradation of inhibitor of kappaB, thereby blocking the translocation of nuclear factor (NF) kappaB to the nucleus. OmpA+ E. coli-infected cells, subsequently subjected to lipopolysaccharide challenge, were crippled severely in their ability to activate NF-kappaB to induce cytokine/chemokine production. Selective inhibitors of the extracellular signal-regulated kinase (ERK) 1/2 pathway and p38 mitogen-activated protein kinase (MAPK), but not Jun N-terminal kinase, significantly reduced the activation of NF-kappaB and the production of cytokines and chemokines induced by OmpA- E. coli, indicating a role for these kinases in the NF-kappaB/cytokine pathway. It is interesting that the phosphorylation of ERK 1/2 and p38 MAPK was notably reduced in monocytes infected with OmpA+ E. coli when compared with monocytes infected with OmpA- E. coli, suggesting that the modulation of upstream events common for NF-kappaB and MAPKs by the bacterium is possible. The ability of OmpA+ E. coli K1 to inhibit the macrophage response temporarily may enable bacterial survival and growth within the host for the onset of meningitis by E. coli K1. PMID- 15894583 TI - Identification of CCR2, flotillin, and gp49B genes as new G-CSF targets during neutrophilic differentiation. AB - Granulocyte colony-stimulating factor (G-CSF) is a cytokine that stimulates myeloid progenitor cells to proliferate and differentiate into neutrophilic granulocytes. To identify genes induced by G-CSF during neutrophil differentiation, interleukin-3-dependent murine myeloid precursor FDC-P1 cells expressing the G-CSF receptor were stimulated with G-CSF, and the gene expression profile was characterized by DNA microarray analysis. In addition to known signal transducer and activator of transcription-3 target genes, such as suppressor of cytokine signaling-3 (SOCS3), JunB, and p19(INK4D), we newly identified several G CSF targets, including genes for the CC chemokine receptor-2 (CCR2), raft proteins flotillin-1 and flotillin-2, and immunoglobulin-like receptor gp49B. Real-time, quantitative polymerase chain reaction analyses revealed that the expression of these genes was induced in various myeloid cell lines by G-CSF. Furthermore, when HoxA9-immortalized bone marrow progenitors were induced by G CSF to differentiate into mature neutrophils, all of these genes were strongly activated. These genes could be categorized into three groups based on their time course of expression: immediate-early (approximately 20 min, SOCS3), mid-early (2 4 h, flotillin-1/2 and gp49B), and late (>12 h, CCR2). This suggests that different transcriptional mechanisms are involved in the regulation of these genes. We show that bone marrow neutrophils express functional CCR2, which suggest that CC chemokines may play previously unknown roles in neutrophil activation and chemotaxis. PMID- 15894584 TI - Interferon-gamma-activated STAT-1alpha suppresses MMP-9 gene transcription by sequestration of the coactivators CBP/p300. AB - Interferon-gamma (IFN-gamma) is a pleiotropic cytokine involved in aspects of immune regulation, cell proliferation, and host defense mechanisms directed toward various cancers. Some of the biological functions of IFN-gamma are achieved through inhibition of gene expression, although the mechanisms by which IFN-gamma suppresses gene transcription are poorly understood. Herein, we demonstrate the molecular basis by which IFN-gamma mediates suppression of the matrix metalloproteinase-9 (MMP-9) gene. IFN-gamma-activated signal transducer and activator of transcription-1alpha (STAT-1alpha) suppresses MMP-9 gene transcription, which is dependent on phosphorylation of tyrosine 701 but not phosphorylation of serine 727. The coactivator cyclic AMP response element binding protein-binding protein (CBP) is an important component of induction of MMP-9 gene transcription. IFN-gamma induces the in vivo association of STAT 1alpha and CBP and decreases the association of CBP to the MMP-9 promoter. IFN gamma does not influence the stability of CBP nor does IFN-gamma affect chromatin remodeling events on the MMP-9 promoter. IFN-gamma inhibits the assembly of the MMP-9 transcription complex by suppressing H3/H4 acetylation and inhibiting recruitment of Pol II to the MMP-9 promoter. These findings indicate that IFN gamma/STAT-1alpha exert their inhibitory effects by affecting multiple aspects of MMP-9 gene transcription. PMID- 15894585 TI - Polysaccharide purified from Ganoderma lucidum induced activation and maturation of human monocyte-derived dendritic cells by the NF-kappaB and p38 mitogen activated protein kinase pathways. AB - Ganoderma lucidum, a fungus native to China, has been widely used to promote health and longevity in the Chinese. The polysaccharide component with a branched (1-->6)-beta-D-glucan moiety of G. lucidum (PS-G) has been reported to exert anti tumor activity and activation of natural killer cells. In this study, we investigated the effects of PS-G on human monocyte-derived dendritic cells (DC). Treatment of DC with PS-G resulted in the enhanced cell-surface expression of CD80, CD86, CD83, CD40, CD54, and human leukocyte antigen (HLA)-DR, as well as the enhanced production of interleukin (IL)-12p70, p40, and IL-10 and also IL 12p35, p40, and IL-10 mRNA expression, and the capacity for endocytosis was suppressed in DC. In addition, treatment of DC with PS-G resulted in enhanced T cell-stimulatory capacity and increased T cell secretion of interferon-gamma and IL-10. Neutralization with antibodies against Toll-like receptor (TLR)-4 inhibited the PS-G-induced production of IL-12 p40 and IL-10, suggesting a vital role for TLR-4 in signaling DC upon incubation with PS-G. Further study showed that PS-G was able to augment inhibitor of kappaB (IkappaB) kinase and nuclear factor (NF)-kappaB activity and also IkappaB alpha and p38 mitogen-activated protein kinase (MAPK) phosphorylation. Further, inhibition of NF-kappaB by helenalin and p38 MAPK by SB98059 prevented the effects of PS-G in the expression of CD80, CD86, CD83, CD40, CD54, and HLA-DR and production of IL-12p70, p40, and IL-10 in various degrees. Taken together, our data demonstrate that PS-G can effectively promote the activation and maturation of immature DC, suggesting that PS-G may possess a potential in regulating immune responses. PMID- 15894586 TI - T cell homeostasis in tolerance and immunity. AB - The size of the peripheral T cell pool is remarkably stable throughout life, reflecting precise regulation of cellular survival, proliferation, and apoptosis. Homeostatic proliferation refers to the process by which T cells spontaneously proliferate in a lymphopenic host. The critical signals driving this expansion are "space," contact with self-major histocompatibility complex (MHC)/peptide complexes, and cytokine stimulation. A number of studies have delineated an association between T cell lymphopenia, compensatory homeostatic expansion, and the development of diverse autoimmune syndromes. In the nonobese diabetic mouse model of type 1 diabetes, lymphopenia-induced homeostatic expansion fuels the generation of islet-specific T cells. Excess interleukin-21 facilitates T cell cycling but limited survival, resulting in recurrent stimulation of T cells specific for self-peptide/MHC complexes. Indeed, data from several experimental models of autoimmunity indicate that a full T cell compartment restrains homeostatic expansion of self-reactive cells that could otherwise dominate the repertoire. This review describes the mechanisms that govern T cell homeostatic expansion and outlines the evidence that lymphopenia presents a risk for development of autoimmune disease. PMID- 15894587 TI - Increased Fas ligand expression of CD4+ T cells by HCV core induces T cell dependent hepatic inflammation. AB - Hepatitis C virus (HCV) infection is associated with a high rate of viral persistence and the development of chronic liver disease. The expression of HCV core protein in T cells has previously been reported to alter T cell activation and has been linked to the development of liver inflammation. However, the molecular and cellular basis for the role of HCV core-expressing T cells in liver inflammation is not understood. Here, using double-transgenic mice of CD2/HCV core transgenic mice and ovalbumin (OVA)-specific T cell receptor transgenic mice, we demonstrated that in vivo antigenic stimulation (OVA peptide administration) triggers a marked influx of core-expressing, antigen-specific, transgenic CD4+ T cells into the liver of these mice. Phenotypic analysis of the liver-infiltrating T cells revealed high expression levels of CD44 and Fas ligand (FasL). Adoptive transfer of liver-infiltrating, core-expressing CD4+ T cells into severe combined immunodeficiency mice directly demonstrated the capacity of these activated T cells to induce liver inflammation. It is important that anti FasL antibody treatment of the mice at the time of cell transfer abrogated the liver inflammation induced by core-expressing CD4+ T cells. These findings suggest that activated T lymphocytes expressing elevated levels of FasL may be involved in the bystander killing of hepatocyte, as well as the induction of chronic liver inflammation, by promoting recruitment of proinflammatory cells to the liver. PMID- 15894588 TI - Impaired inflammatory angiogenesis, but not leukocyte influx, in mice lacking TNFR1. AB - The majority of biological responses classically attributed to tumor necrosis factor alpha (TNF-alpha) is mediated by p55 receptor (TNFR1). Here, we aimed to clarify the biological role of TNFR1-mediated signals in an in vivo inflammatory angiogenesis model. Polyester-polyurethane sponges, used as a framework for tissue growth, were implanted in C57Bl/6 mice. These implants were collected at days 1, 7, and 14 post-implant for enzyme-linked immunosorbent assay or at days 7 and 14 for hemoglobin, myeloperoxidase, and N-acetylglucosaminidase measurements, used as indexes for angiogenesis, neutrophil, and macrophage accumulation, respectively. In TNFR1-deficient C57Bl/6 mice, there was a significant decrease in sponge vascularization but not in late inflammatory cell influx. It is interesting that levels of vascular endothelial growth factor were significantly lower in TNFR1-deficient than in wild-type mice at days 1 and 7. Levels of angiogenic chemokines, CC chemokine ligand 2/murine homologue of monocyte chemoattractant protein-1 and CXC chemokine ligand 1-3/keratinocyte-derived chemokine, were significantly lower in TNFR1-deficient mice at days 1 and 7 after implantation, respectively. These observations suggest that TNFR1-mediated signals have a critical role in sponge-induced angiogenesis, possibly by influencing the effector state of inflammatory cells and hence, modulating the angiogenic molecular network. PMID- 15894589 TI - Sphingosine 1-phosphate receptor agonist FTY720-phosphate causes marginal zone B cell displacement. AB - FTY720 is an immunosuppressive agent that modulates lymphocyte trafficking. It is phosphorylated in vivo to FTY720-phosphate (FTY-P) and binds to a family of G protein-coupled receptors recognizing sphingosine 1-phosphate (S1P) as the natural ligand. It has previously been reported that FTY-P blocks egress of lymphocytes from the thymus and lymph nodes, resulting in peripheral blood lymphopenia. We now report that FTY-P also causes displacement of marginal zone (MZ) B cells to the splenic follicles, an effect that is similar to that observed after in vivo administration of lipopolysaccharide. This effect is specific to B cells in the MZ, as treatment with FTY-P does not cause redistribution of the resident macrophage population. A small but statistically significant decrease in the expression of beta1 integrin on MZ B cells was observed with FTY-P treatment. The redistribution of MZ B cells from the MZ sinuses does not abolish the ability of these cells to respond to the T-independent antigen, trinitrophenol-Ficoll. It has been proposed that the displacement of MZ B cells to the follicles is an indication of cell activation. Consistent with this, FTY-P caused an increase in percentage of MZ B cells expressing activation markers CD9, CD1d, and CD24. These results suggest that S1P receptors on MZ B cells are responsible for their mobilization to follicles. PMID- 15894590 TI - The mucosal immune system: from control of inflammation to protection against infections. AB - The IV meeting of the European Mucosal Immunology Group, held October 8-10, 2004, in Lyon, gathered fundamental and clinical research scientists to discuss the most recent updates on basic and clinical aspects of mucosal immunology. The meeting was focused on innate and acquired immune mechanisms underlying handling and immune recognition of commensals, allergens, and pathogens by the mucosal immune system and its outcome in health and disease as well as for vaccine development. The scientific program featured five topics of growing interest for fundamental research scientists and clinicians, including the role of commensal bacteria in mucosal immunity; function of dendritic cells in infection, inflammation, and tolerance; control of mucosal inflammation by regulatory T cells; novel routes and adjuvants for mucosal vaccines; and mucosal immunity against HIV infection and vaccination strategies. PMID- 15894591 TI - Cohort Profile: the Australian Longitudinal Study on Women's Health. PMID- 15894592 TI - Maternal and childhood nutrition and later blood pressure levels in young Guatemalan adults. AB - BACKGROUND: Low birth weight and subsequent rapid child growth are associated with later blood pressure levels. The role of maternal and child nutrition in this association remains unclear. METHODS: We studied 450 men and women (ages 21 29 years) born during a randomized trial of protein-energy supplementation (Atole) vs low energy/no protein supplementation (Fresco) in pregnancy and early childhood in four rural Guatemalan villages from 1969 to 1977. RESULTS: Protein energy supplementation was not associated with differences in blood pressure in adulthood (diastolic blood pressure (DBP): beta = 0.69 mm Hg, 95% confidence internal (CI) (20.82-2.19); P = 0.37; systolic blood pressure (SBP): beta = 0.17 mm Hg, 95% CI (21.68-2.02); P = 0.86). Within the Atole group, maternal height was associated with later SBP (0.22 mm Hg/cm, 95% CI (20.002-0.45); P = 0.05). No other associations between maternal nutritional status, birth size, child growth, or supplement intake were observed for adult blood pressure. CONCLUSIONS: Our data do not support the role of maternal nutrition during pregnancy, birth size, or early child growth in programming adult blood pressure. Likewise, we found no effect of protein-energy supplementation in pregnancy or in early childhood on blood pressure in young adults. PMID- 15894593 TI - Trends in primary care consultations for asthma in Switzerland, 1989-2002. AB - BACKGROUND: There is widespread debate about trends in the occurrence of asthma in industrialized countries. This study was conducted to investigate time trends in consultations for asthma in primary care in Switzerland. METHODS: Prospective observational study from 1989 to 2002 within the Swiss Sentinel Surveillance Network; a primary care surveillance system. We used time series analysis and non parametric smoothing methods to investigate long-term and short-term trends in rates of asthma episodes per 1000 consultations. From 1994 to 2002 we compared rates of first episodes with all subsequent consultations for asthma. RESULTS: Overall consultation rates for asthma per 1000 primary care consultations increased from 1989 to 1994 then stabilized and have declined since 2000. Long term trends showed a small decline in first consultations for asthma from an average of 0.78 (95% credibility intervals (CI) 0.74-0.81) in 1999 to 0.62 (95% CI 0.55-0.69) per 1000 consultations in 2002. Subsequent consultations for asthma have been declining since at least 1994, from an average of 1.5 (95% CI 1.40 1.61) per 1000 consultations in 1994 to 0.93 (95% CI 0.82-1.04) in 2002. In addition, the ratio of subsequent to first episodes of asthma fell in all age groups. CONCLUSIONS: In Switzerland, primary care consultations for asthma, subsequent to the initial diagnosis, have been declining since 1994. This is more likely to be owing to an increase in the use of home medication than to a shift in care to hospital settings. The incidence of diagnosed asthma might also be decreasing. PMID- 15894594 TI - POMT2 mutations cause alpha-dystroglycan hypoglycosylation and Walker-Warburg syndrome. AB - BACKGROUND: Walker-Warburg syndrome (WWS) is an autosomal recessive condition characterised by congenital muscular dystrophy, structural brain defects, and eye malformations. Typical brain abnormalities are hydrocephalus, lissencephaly, agenesis of the corpus callosum, fusion of the hemispheres, cerebellar hypoplasia, and neuronal overmigration, which causes a cobblestone cortex. Ocular abnormalities include cataract, microphthalmia, buphthalmos, and Peters anomaly. WWS patients show defective O-glycosylation of alpha-dystroglycan (alpha-DG), which plays a key role in bridging the cytoskeleton of muscle and CNS cells with extracellular matrix proteins, important for muscle integrity and neuronal migration. In 20% of the WWS patients, hypoglycosylation results from mutations in either the protein O-mannosyltransferase 1 (POMT1), fukutin, or fukutin related protein (FKRP) genes. The other genes for this highly heterogeneous disorder remain to be identified. OBJECTIVE: To look for mutations in POMT2 as a cause of WWS, as both POMT1 and POMT2 are required to achieve protein O mannosyltransferase activity. METHODS: A candidate gene approach combined with homozygosity mapping. RESULTS: Homozygosity was found for the POMT2 locus at 14q24.3 in four of 11 consanguineous WWS families. Homozygous POMT2 mutations were present in two of these families as well as in one patient from another cohort of six WWS families. Immunohistochemistry in muscle showed severely reduced levels of glycosylated alpha-DG, which is consistent with the postulated role for POMT2 in the O-mannosylation pathway. CONCLUSIONS: A fourth causative gene for WWS was uncovered. These genes account for approximately one third of the WWS cases. Several more genes are anticipated, which are likely to play a role in glycosylation of alpha-DG. PMID- 15894595 TI - Pseudoxanthoma elasticum: a clinical, pathophysiological and genetic update including 11 novel ABCC6 mutations. AB - Pseudoxanthoma elasticum (PXE) is an inherited systemic disease of connective tissue primarily affecting the skin, retina, and cardiovascular system. It is characterised pathologically by elastic fibre mineralisation and fragmentation (so called "elastorrhexia"), and clinically by high heterogeneity in age of onset and the extent and severity of organ system involvement. PXE was recently associated with mutations in the ABCC6 (ATP binding cassette subtype C number 6) gene. At least one ABCC6 mutation is found in about 80% of patients. These mutations are identifiable in most of the 31 ABCC6 exons and consist of missense, nonsense, frameshift mutations, or large deletions. No correlation between the nature or location of the mutations and phenotype severity has yet been established. Recent findings support exclusive recessive inheritance. The proposed prevalence of PXE is 1/25,000, but this is probably an underestimate. ABCC6 encodes the protein ABCC6 (also known as MRP6), a member of the large ATP dependent transmembrane transporter family that is expressed predominantly in the liver and kidneys, and only to a lesser extent in tissues affected by PXE. The physiological substrates of ABCC6 remain to be determined, but the current hypothesis is that PXE should be considered to be a metabolic disease with undetermined circulating molecules interacting with the synthesis, turnover, or maintenance of elastic fibres. PMID- 15894596 TI - Nine unknown rearrangements in 16p13.3 and 11p15.4 causing alpha- and beta thalassaemia characterised by high resolution multiplex ligation-dependent probe amplification. AB - BACKGROUND: Approximately 80% of the alpha- and 10% of the beta-thalassaemias are caused by genomic deletions involving the alpha- and beta-globin gene clusters on chromosomes 16p13.3 and 11p15.5, respectively. Gap-PCR, Southern blot analysis, and fluorescent in situ hybridisation are commonly used to identify these deletions; however, many deletions go undetected using conventional techniques. METHODS: Patient samples for which no abnormalities had been found using conventional DNA techniques were analysed by a three colour multiplex ligation dependent probe amplification assay. Two sets of 35 and 50 probes, covering a region of 700 kb of the alpha- and 500 kb of the beta-globin gene cluster, respectively, were designed to detect rearrangements in the alpha- and beta globin gene clusters. RESULTS: In 19 out of 38 patient samples, we found 11 different alpha-thalassaemia deletions, six of which were not previously described. Two novel deletions leaving the alpha-globin gene cluster intact were found to cause a complete downregulation of the downstream alpha-genes. Similarly, 31 out of 51 patient samples were found to carry 10 different deletions involving the beta-globin gene cluster, three of which were not previously described. One involves the deletion of the locus control region leaving the beta-globin gene cluster intact. CONCLUSIONS: These deletions, which are not easily detected by conventional techniques, may have clinical implications during pregnancy ranging from mild to life threatening microcytic haemolytic anaemia in neonates. The approach as described here is a rapid and sensitive method for high resolution analysis of the globin gene clusters and for any region of the genome. PMID- 15894597 TI - Camurati-Engelmann disease: review of the clinical, radiological, and molecular data of 24 families and implications for diagnosis and treatment. AB - Camurati-Engelmann disease (CED) is a rare autosomal dominant type of bone dysplasia. This review is based on the unpublished and detailed clinical, radiological, and molecular findings in 14 CED families, comprising 41 patients, combined with data from 10 other previously reported CED families. For all 100 cases, molecular evidence for CED was available, as a mutation was detected in TGFB1, the gene encoding transforming growth factor (TGF) beta1. Pain in the extremities was the most common clinical symptom, present in 68% of the patients. A waddling gait (48%), easy fatigability (44%), and muscle weakness (39%) were other important features. Radiological symptoms were not fully penetrant, with 94% of the patients showing the typical long bone involvement. A large percentage of the patients also showed involvement of the skull (54%) and pelvis (63%). The review provides an overview of possible treatments, diagnostic guidelines, and considerations for prenatal testing. The detailed description of such a large set of CED patients will be of value in establishing the correct diagnosis, genetic counselling, and treatment. PMID- 15894598 TI - High-resolution computed tomography in idiopathic pulmonary fibrosis: diagnosis and prognosis. AB - RATIONALE: High-resolution computed tomography (HRCT) is an integral aspect of the evaluation of patients with suspected idiopathic pulmonary fibrosis (IPF). However, few studies have evaluated its use in a large cohort. OBJECTIVES: To describe HRCT features in patients with mild to moderate IPF, compare diagnostic evaluations by a radiology core (three thoracic radiologists) with those by study site radiologists, correlate baseline clinical and physiologic variables with HRCT findings, and evaluate their association with mortality. METHODS: We assessed HRCT scans from patients with IPF (n = 315) enrolled in a randomized controlled study evaluating IFN-gamma1b. MEASUREMENTS AND MAIN RESULTS: There was concordance between study-site and core radiologists regarding the diagnosis of IPF in 86% of cases. Diffusing capacity of carbon monoxide (DLCO) was the physiologic characteristic most highly correlated with HRCT findings. Multivariate analysis identified three independent predictors of mortality: a higher extent of fibrosis score increased the risk of death (p < 0.0001), whereas a higher percent-predicted DLCO (p = 0.004) and treatment assignment to IFN gamma1b rather than placebo (p = 0.04) reduced the risk of death. CONCLUSIONS: A study-site diagnosis of IPF on HRCT was regularly confirmed by core radiologists. Extent of reticulation and honeycombing on HRCT is an important independent predictor of mortality in patients with IPF. PMID- 15894599 TI - Interleukin-10 gene expression in acute virus-induced asthma. AB - RATIONALE: Virus-induced asthma is characterized by marked neutrophil influx and eosinophil degranulation, suggesting a mode of immunopathogenesis different from that of allergen-induced asthma. OBJECTIVES: This study compared induced sputum cytokine responses in subjects with severe asthma exacerbation and respiratory virus infection with those of patients with stable asthma, healthy control subjects, and virus-infected nonasthmatic subjects. METHODS: Subject infection status and pulmonary history were established on the basis of common cold and asthma questionnaires, and lung function and atopy tests were performed. Respiratory virus infection was diagnosed by cell culture and direct polymerase chain reaction, using induced sputum. The induced sputum cellular profile was examined and cytokine gene expression was assessed by quantitative real-time polymerase chain reaction. RESULTS: A respiratory virus was detected in 78% of subjects with acute asthma. Specific viruses detected were rhinovirus (83%), influenza (15%), enterovirus (4%), and respiratory syncytial virus (2%). Virus infected subjects with acute asthma or no asthma had increased RANTES (regulated on activation, normal T cell expressed and secreted) and macrophage inflammatory protein-1alpha messenger RNAs compared with other groups. Interleukin (IL)-10 mRNA was significantly increased in virus-infected acute asthma and reduced on recovery from acute asthma. IL-5, eotaxin, and IL-8 mRNA transcripts were similar across groups. CONCLUSIONS: Asthma exacerbation triggered by respiratory virus infection is characterized by increased IL-10 gene expression that may explain the suppressed eosinophil influx in acute asthma. Airway neutrophilia due to respiratory virus infection is associated with chemokine gene expression involving RANTES and macrophage inflammatory protein-1alpha. PMID- 15894600 TI - Voltaire, Walpole, and Pasteur: variations on the theme of discovery. PMID- 15894601 TI - Physiologic, biochemical, and imaging characterization of acute lung injury in mice. AB - RATIONALE: Most models of acute lung injury in mice have yet to be fully characterized. OBJECTIVES: To directly compare and contrast endotoxin and oleic acid models of acute lung injury in mice in terms of their physiologic, biochemical, histopathologic, and imaging manifestations. METHODS: Survival studies, lung weights, x-ray computed tomographic scanning, light and electron microscopy, bronchoalveolar lavage, lung uptake of ((18)F)fluorodeoxyglucose, tissue myeloperoxidase, arterial blood gases, mean arterial pressure, and lung tissue prostanoids were measured in separate groups of C57Bl/6 mice (normal animals, endotoxin only [20 microg/g], oleic acid only [0.15 microl/g], or endotoxin + oleic acid). RESULTS: Endotoxin alone caused only mild pulmonary neutrophilic inflammation with little functional or structural damage to the alveolar architecture. In contrast, oleic acid caused severe alveolar damage with the development of alveolar edema of the increased-permeability type with associated abnormalities in gas exchange. When given together, endotoxin and oleic acid acted synergistically to increase pulmonary edema and to worsen gas exchange and hemodynamics, thereby increasing mortality. This synergism was significantly attenuated by the prior administration of the endotoxin antagonist E5564 (eritoran). CONCLUSIONS: Under the conditions of these studies, only mice exposed to oleic acid showed both structural and functional characteristics of acute lung injury. Nevertheless, endotoxin had potent synergistic physiologic effects that increased mortality. Overall, these models, which can be translated to genetically altered mice, are amenable to study with state-of-the-art imaging techniques, and with experimental interventions that can probe the underlying mechanisms of injury. PMID- 15894602 TI - Pretransplant lung function, respiratory failure, and mortality after stem cell transplantation. AB - RATIONALE: The role of pulmonary function before stem cell transplant as a potential risk factor for the development of early post-transplant respiratory failure and mortality is controversial. METHODS: We conducted a retrospective analysis of the pretransplant pulmonary function of 2,852 patients who received their transplant between 1990 and 2001. MEASUREMENTS: Pretransplant FEV(1), FVC, total lung capacity (TLC), diffusing capacity of carbon monoxide (DL(CO)), and the alveolar-arterial oxygen tension difference P(A-a)O(2) were measured and assessed for association with development of early respiratory failure and mortality in Cox proportional hazard logistic models. MAIN RESULTS: In multivariate analyses, progressive decrease of all lung function parameters was associated with a stepwise increase in risk of developing early respiratory failure and mortality when assessed in independent models. On the basis of a significant correlation between FEV(1) and FVC (r = 0.81), FEV(1) and TLC (r = 0.61), and FVC and TLC (r = 0.80), and a lack of correlation between FEV(1) and DL(CO), we developed a pretransplant lung function score based on pretransplant FEV(1) and DL(CO) to determine the extent of pulmonary compromise before transplant. Multivariate analysis indicated that higher pretransplant lung function scores are associated with a significant increased risk for developing early respiratory failure (category II hazard ratio [HR], 1.4; category III HR, 2.2; category IV HR, 3.1; p < 0.001) and death (category II HR, 1.2; category III HR, 2.2; category IV HR, 2.7; p < 0.005). CONCLUSIONS: These results suggest that not only does compromised pretransplant lung function contribute to the risk for development of early respiratory failure and mortality but this risk may be estimated before transplant by grading the extent of FEV(1) and DL(CO) compromise. PMID- 15894603 TI - Prognostic value of bronchiolitis obliterans syndrome stage 0-p in single-lung transplant recipients. AB - RATIONALE: Early diagnosis of bronchiolitis obliterans syndrome (BOS) is critical in understanding pathogenesis and devising therapeutic trials. Although potential BOS stage (BOS 0-p), encompassing early changes in FEV(1) and forced expiratory flow, midexpiratory phase (FEF(25-75%)), has been proposed, there is a paucity of data validating its utility in single-lung transplantation. OBJECTIVE: The aim of this study was to define the predictive ability of BOS 0-p in single-lung transplantation. METHODS: We retrospectively analyzed spirometric data for 197 single-lung recipients. Sensitivity, specificity, and positive predictive value of BOS 0-p were examined over time using Kaplan-Meier methodology. RESULTS: BOS 0 p FEV(1) was associated with higher sensitivity, specificity, and positive predictive value than the FEF(25-75%) criterion over different time periods investigated. The probability of testing positive for BOS 0-p FEV(1) in patients with BOS (sensitivity) was 71% at 2 years before the onset of BOS. The probability of being free from development of BOS 0-p FEV(1) in patients free of BOS at follow-up (specificity) was 93% within the last year. Of patients who met the BOS 0-p FEV(1) criterion, 81% developed BOS or died within 3 years. The specificity and positive predictive value curves for the BOS 0-p FEV(1) were significantly different between patients with underlying restrictive versus obstructive physiology (p = 0.05 and 0.01, respectively). CONCLUSION: The FEV(1) criterion for BOS 0-p provides useful predictive information regarding the risk of development of BOS or death in single-lung recipients. The predictive value of this criterion is higher in patients with underlying restriction and is superior to the FEF(25-75%) criterion. PMID- 15894604 TI - 5-Lipoxygenase deficiency prevents respiratory failure during ventilator-induced lung injury. AB - RATIONALE: Mechanical ventilation with high VT (HVT) progressively leads to lung injury and decreased efficiency of gas exchange. Hypoxic pulmonary vasoconstriction (HPV) directs blood flow to well-ventilated lung regions, preserving systemic oxygenation during pulmonary injury. Recent experimental studies have revealed an important role for leukotriene (LT) biosynthesis by 5 lipoxygenase (5LO) in the impairment of HPV by endotoxin. OBJECTIVES: To investigate whether or not impairment of HPV contributes to the hypoxemia associated with HVT and to evaluate the role of LTs in ventilator-induced lung injury. METHODS: We studied wild-type and 5LO-deficient mice ventilated for up to 10 hours with low VT (LVT) or HVT. RESULTS: In wild-type mice, HVT, but not LVT, increased pulmonary vascular permeability and edema formation, impaired systemic oxygenation, and reduced survival. HPV, as reflected by the increase in left pulmonary vascular resistance induced by left mainstem bronchus occlusion, was markedly impaired in animals ventilated with HVT. HVT ventilation increased bronchoalveolar lavage levels of LTs and neutrophils. In 5LO-deficient mice, the HVT-induced increase of pulmonary vascular permeability and worsening of respiratory mechanics were markedly attenuated, systemic oxygenation was preserved, and survival increased. Moreover, in 5LO-deficient mice, HVT ventilation did not impair the ability of left mainstem bronchus occlusion to increase left pulmonary vascular resistance. Administration of MK886, a 5LO activity inhibitor, or MK571, a selective cysteinyl-LT(1) receptor antagonist, largely prevented ventilator-induced lung injury. CONCLUSIONS: These results indicate that LTs play a central role in the lung injury and impaired oxygenation induced by HVT ventilation. PMID- 15894605 TI - Oxidative stress in pulmonary fibrosis: a possible role for redox modulatory therapy. AB - Idiopathic ulmonary fibrosis (histopathology of usual interstitial pneumonia) is a progressive lung disease of unknown etiology. No treatment has been shown to improve the prognosis of the patients with this disease. Recent evidence, including the observations that the patients with idiopathic pulmonary fibrosis have higher levels of oxidant stress than control patients, and a recent multicenter European study examining the effect of the antioxidant N acetylcysteine on the progression of idiopathic pulmonary fibrosis suggest that the cellular redox state may play a significant role in the progression of this disease. These complex mechanisms include activation of growth factors as well as regulation of matrix metalloproteinases and protease inhibitors. Potential future approaches for the therapy of interstitial pulmonary fibrosis may involve synthetic agents able to modulate cellular redox state. Investigation into therapeutic approaches to inhibit oxidant-mediated reactions in the initiation and progression of pulmonary fibrosis may provide hope for the future treatment of this disease. PMID- 15894606 TI - Three-dimensional structure of a halotolerant algal carbonic anhydrase predicts halotolerance of a mammalian homolog. AB - Protein molecular adaptation to drastically shifting salinities was studied in dCA II, an alpha-type carbonic anhydrase (EC 4.2.1.1) from the exceptionally salt tolerant unicellular green alga Dunaliella salina. The salt-inducible, extracellular dCA II is highly salt-tolerant and thus differs from its mesophilic homologs. The crystal structure of dCA II, determined at 1.86-A resolution, is globally similar to other alpha-type carbonic anhydrases except for two extended alpha-helices and an added Na-binding loop. Its unusual electrostatic properties include a uniformly negative surface electrostatic potential of lower magnitude than that observed in the highly acidic halophilic proteins and an exceptionally low positive potential at a site adjoining the catalytic Zn(2+) compared with mesophilic homologs. The halotolerant dCA II also differs from typical halophilic proteins in retaining conformational stability and solubility in low to high salt concentrations. The crucial role of electrostatic features in dCA II halotolerance is strongly supported by the ability to predict the unanticipated halotolerance of the murine CA XIV isozyme, which was confirmed biochemically. A proposal for the functional significance of the halotolerance of CA XIV in the kidney is presented. PMID- 15894607 TI - Systems analysis of transcriptome and proteome in retinoic acid/arsenic trioxide induced cell differentiation/apoptosis of promyelocytic leukemia. AB - Understanding the complexity and dynamics of cancer cells in response to effective therapy requires hypothesis-driven, quantitative, and high-throughput measurement of genes and proteins at both spatial and temporal levels. This study was designed to gain insights into molecular networks underlying the clinical synergy between retinoic acid (RA) and arsenic trioxide (ATO) in acute promyelocytic leukemia (APL), which results in a high-quality disease-free survival in most patients after consolidation with conventional chemotherapy. We have applied an approach integrating cDNA microarray, 2D gel electrophoresis with MS, and methods of computational biology to study the effects on APL cell line NB4 treated with RA, ATO, and the combination of the two agents and collected in a time series. Numerous features were revealed that indicated the coordinated regulation of molecular networks from various aspects of granulocytic differentiation and apoptosis at the transcriptome and proteome levels. These features include an array of transcription factors and cofactors, activation of calcium signaling, stimulation of the IFN pathway, activation of the proteasome system, degradation of the PML-RARalpha oncoprotein, restoration of the nuclear body, cell-cycle arrest, and gain of apoptotic potential. Hence, this investigation has provided not only a detailed understanding of the combined therapeutic effects of RA/ATO in APL but also a road map to approach hematopoietic malignancies at the systems level. PMID- 15894608 TI - On the orientation of the backbone dipoles in native folds. AB - The role of electrostatic interactions in determining the native fold of proteins has been investigated by analyzing the alignment of peptide bond dipole moments with the local electrostatic field generated by the rest of the molecule with and without solvent effects. This alignment was calculated for a set of 112 native proteins by using charges from a gas phase potential. Most of the peptide dipoles in this set of proteins are on average aligned with the electrostatic field. The dipole moments associated with alpha-helical conformations show the best alignment with the electrostatic field, followed by residues in beta-strand conformations. The dipole moments associated with other secondary structure elements are on average better aligned than in randomly generated conformations. The alignment of a dipole with the local electrostatic field depends on both the topology of the native fold and the charge distribution assumed for all of the residues. The influences of (i) solvent effects, (ii) different sets of charges, and (iii) the charge distribution assumed for the whole molecule were examined with a subset of 22 proteins each of which contains <30 ionizable groups. The results show that alternative charge distribution models lead to significant differences among the associated electrostatic fields, whereas the electrostatic field is less sensitive to the particular set of the adopted charges themselves (empirical conformational energy program for peptides or parameters for solvation energy). PMID- 15894609 TI - Physics-based protein-structure prediction using a hierarchical protocol based on the UNRES force field: assessment in two blind tests. AB - Recent improvements in the protein-structure prediction method developed in our laboratory, based on the thermodynamic hypothesis, are described. The conformational space is searched extensively at the united-residue level by using our physics-based UNRES energy function and the conformational space annealing method of global optimization. The lowest-energy coarse-grained structures are then converted to an all-atom representation and energy-minimized with the ECEPP/3 force field. The procedure was assessed in two recent blind tests of protein-structure prediction. During the first blind test, we predicted large fragments of alpha and alpha+beta proteins [60-70 residues with C(alpha) rms deviation (rmsd) <6 A]. However, for alpha+beta proteins, significant topological errors occurred despite low rmsd values. In the second exercise, we predicted whole structures of five proteins (two alpha and three alpha+beta, with sizes of 53-235 residues) with remarkably good accuracy. In particular, for the genomic target TM0487 (a 102-residue alpha+beta protein from Thermotoga maritima), we predicted the complete, topologically correct structure with 7.3-A C(alpha) rmsd. So far this protein is the largest alpha+beta protein predicted based solely on the amino acid sequence and a physics-based potential-energy function and search procedure. For target T0198, a phosphate transport system regulator PhoU from T. maritima (a 235-residue mainly alpha-helical protein), we predicted the topology of the whole six-helix bundle correctly within 8 A rmsd, except the 32 C-terminal residues, most of which form a beta-hairpin. These and other examples described in this work demonstrate significant progress in physics-based protein-structure prediction. PMID- 15894610 TI - The elongata mutants identify a functional Elongator complex in plants with a role in cell proliferation during organ growth. AB - The key enzyme for transcription of protein-encoding genes in eukaryotes is RNA polymerase II (RNAPII). The recruitment of this enzyme during transcription initiation and its passage along the template during transcription elongation is regulated through the association and dissociation of several complexes. Elongator is a histone acetyl transferase complex, consisting of six subunits (ELP1-ELP6), that copurifies with the elongating RNAPII in yeast and humans. We demonstrate that point mutations in three Arabidopsis thaliana genes, encoding homologs of the yeast Elongator subunits ELP1, ELP3 (histone acetyl transferase), and ELP4 are responsible for the phenotypes of the elongata2 (elo2), elo3, and elo1 mutants, respectively. The elo mutants are characterized by narrow leaves and reduced root growth that results from a decreased cell division rate. Morphological and molecular phenotypes show that the ELONGATA (ELO) genes function in the same biological process and the epistatic interactions between the ELO genes can be explained by the model of complex formation in yeast. Furthermore, the plant Elongator complex is genetically positioned in the process of RNAPII-mediated transcription downstream of Mediator. Our data indicate that the Elongator complex is evolutionarily conserved in structure and function but reveal that the mechanism by which it stimulates cell proliferation is different in yeast and plants. PMID- 15894611 TI - High-resolution x-ray crystal structures of the villin headpiece subdomain, an ultrafast folding protein. AB - The 35-residue subdomain of the villin headpiece (HP35) is a small ultrafast folding protein that is being intensely studied by experiments, theory, and simulations. We have solved the x-ray structures of HP35 and its fastest folding mutant [K24 norleucine (nL)] to atomic resolution and compared their experimentally measured folding kinetics by using laser temperature jump. The structures, which are in different space groups, are almost identical to each other but differ significantly from previously solved NMR structures. Hence, the differences between the x-ray and NMR structures are probably not caused by lattice contacts or crystal/solution differences, but reflect the higher accuracy of the x-ray structures. The x-ray structures reveal important details of packing of the hydrophobic core and some additional features, such as cross-helical H bonds. Comparison of the x-ray structures indicates that the nL substitution produces only local perturbations. Consequently, the finding that the small stabilization by the mutation is completely reflected in an increased folding rate suggests that this region of the protein is as structured in the transition state as in the folded structure. It is therefore a target for engineering to increase the folding rate of the subdomain from approximately 0.5 micros(-1) for the nL mutant to the estimated theoretical speed limit of approximately 3 micros( 1). PMID- 15894612 TI - The activating NKG2D receptor assembles in the membrane with two signaling dimers into a hexameric structure. AB - The activating NKG2D receptor plays a critical role in innate and adaptive immune responses by natural killer cells and subpopulations of T cells. The human receptor assembles with the DAP10 signaling dimer, and it is thought that one NKG2D homodimer pairs with a single DAP10 dimer by formation of two salt bridges between charged transmembrane (TM) residues. However, direct stoichiometry measurements demonstrated that one NKG2D homodimer assembles with four DAP10 chains. Selective mutation of one of the basic TM residues of NKG2D resulted in loss of two DAP10 chains, indicating that each TM arginine serves as an interaction site for a DAP10 dimer. Assembly of the hexameric structure was cooperative because this mutation also significantly reduced NKG2D dimerization. A monomeric NKG2D TM peptide was sufficient for assembly with a DAP10 dimer, indicating that the interaction between these proteins occurs in the membrane environment. Formation of a three-helix interface among the TM domains involved ionizable residues from all three chains, the TM arginine of NKG2D and both TM aspartic acids of the DAP10 dimer. The organization of the TM domains thus shows similarities to the T cell antigen receptor-CD3 complex, in particular to the six chain assembly intermediate between T cell antigen receptor and the CD3delta epsilon and CD3gamma epsilon dimers. Binding of a single ligand can thus result in phosphorylation of four DAP10 chains, which may be relevant for the sensitivity of NKG2D receptor signaling, in particular in situations of low ligand density. PMID- 15894613 TI - Inhibition of amyloid precursor protein processing by beta-secretase through site directed antibodies. AB - Amyloid-beta peptide (AbetaP) that accumulates in the Alzheimer's diseased brain is derived from proteolytic processing of the amyloid precursor protein (APP) by means of beta- and gamma-secretases. The beta-secretase APP cleaving enzyme (BACE), which generates the N terminus of AbetaP, has become a target of intense research aimed at blocking the enzyme activity, thus reducing AbetaP and, subsequently, plaque formation. The search for specific inhibitors of beta secretase activity as a possible treatment for Alzheimer's disease intensified with the discovery that BACE may be involved in processing other non-APP substrates. The presence of the APP-BACE complex in early endosomes highlights the cell surface as a potential therapeutic target, suggesting that interference in APP-BACE interaction at the cell surface may affect amyloid-beta production. We present here a unique approach to inhibit AbetaP production by means of antibodies against the beta-secretase cleavage site of APP. These antibodies were found to bind human APP overexpressed by CHO cells, and the formed immunocomplex was visualized in the early endosomes. Indeed, blocking of the beta-secretase site by these antibodies interfered with BACE activity and inhibited both intracellular and extracellular AbetaP formation in these cells. PMID- 15894614 TI - CDX1 is an important molecular mediator of Barrett's metaplasia. AB - The molecular pathogenesis of Barrett's metaplasia (BM) of the esophagus is poorly understood. The change to an intestinal phenotype occurs on a background of esophagitis due to refluxing acid and bile. CDX1, an important regulator of normal intestinal development, was studied as a potential key molecule in the pathogenesis of BM. CDX1 mRNA and protein were universally expressed in all samples of BM tested but not in normal esophageal squamous or gastric body epithelia. This tissue-specific expression was attributable to the methylation status of the CDX1 promoter. Conjugated bile salts and the inflammatory cytokines TNF-alpha and IL-1beta were all found to increase CDX1 mRNA expression in vitro. These effects were primarily mediated by NF-kappaB signaling but only occurred when the CDX1 promoter was unmethylated or partially methylated. The data suggest that CDX1 is a key molecule linking etiological agents of BM to the development of an intestinal phenotype. Although the initial trigger for CDX1 promoter demethylation is not yet identified, it seems likely that demethylation of its promoter may be the key to the induction and maintenance of CDX1 expression and so of the BM phenotype. PMID- 15894615 TI - Prion and water: tight and dynamical hydration sites have a key role in structural stability. AB - The propensity to form fibril in disease-related proteins is a widely studied phenomenon, but its correlation, if any, with structural characteristics of the associated proteins is not clearly understood. However, the observation has been made that some proteins that readily form amyloid have a significant number of backbone H bonds that are exposed to solvent molecules, suggesting that these regions have a propensity toward protein interaction and aggregation [Fernandez, A. & Scheraga, H. A. (2003) Proc. Natl. Acad. Sci. USA 100, 113-118]. High resolution x-ray structures of the sheep and human C-terminal prion protein have provided a useful description of surface and partially buried waters. By molecular dynamics simulations, we investigated the structural role of these water molecules. The solvent dynamical behavior on the protein surface reveals significant features about the stability and the potential interactions of the prion protein. The protein presents regions of tightly bound conserved waters that are necessary to hold in place local elements of the fold, as well as regions where the local water is in fast exchange with bulk water. These results are evidenced by a map of the spatial distribution entropy of the solvent around the protein. The particular behavior of the solvent around these regions may be crucial in the folding stability and in terms of aggregation loci. PMID- 15894616 TI - Autophagy promotes MHC class II presentation of peptides from intracellular source proteins. AB - MHC-peptide complexes mediate key functions in adaptive immunity. In a classical view, MHC-I molecules present peptides from intracellular source proteins, whereas MHC-II molecules present antigenic peptides from exogenous and membrane proteins. Nevertheless, substantial crosstalk between these two pathways has been observed. We investigated the influence of autophagy on the MHC-II ligandome and demonstrated that peptide presentation is altered considerably upon induction of autophagy. The presentation of peptides from intracellular and lysosomal source proteins was strongly increased on MHC-II in contrast with peptides from membrane and secreted proteins. In addition, autophagy influenced the MHC-II antigen processing machinery. Our study illustrates a profound influence of autophagy on the class II peptide repertoire and suggests that this finding has implications for the regulation of CD4(+) T cell-mediated processes. PMID- 15894617 TI - Structural basis for lysidine formation by ATP pyrophosphatase accompanied by a lysine-specific loop and a tRNA-recognition domain. AB - Lysidine, a lysine-combined modified cytidine, is exclusively located at the anticodon wobble position (position 34) of eubacterial tRNA(Ile)(2) and not only converts the codon specificity from AUG to AUA, but also converts the aminoacylation specificity from recognition by methionyl-tRNA synthetase to that by isoleucyl-tRNA synthetase (IleRS). Here, we report the crystal structure of lysidine synthetase (TilS) from Aquifex aeolicus at 2.42-A resolution. TilS forms a homodimer, and each subunit consists of the N-terminal dinucleotide-binding fold domain (NTD), with a characteristic central hole, and the C-terminal globular domain (CTD) connected by a long alpha-helical linker. The NTD shares striking structural similarity with the ATP-pyrophosphatase domain of GMP synthetase, which reminds us of the two-step reaction by TilS: adenylation of C34 and lysine attack on the C2 carbon. Conserved amino acid residues are clustered around the NTD central hole. Kinetic analyses of the conserved residues' mutants indicated that C34 of tRNA(Ile)(2) is adenylated by an ATP lying across the NTD central hole and that a lysine, which is activated at a loop appended to the NTD, nucleophilically attacks the C2 carbon from the rear. Escherichia coli TilS (called MesJ) has an additional CTD, which may recognize the tRNA(Ile)(2) acceptor stem. In contrast, a mutational study revealed that A. aeolicus TilS does not recognize the tRNA acceptor stem but recognizes the C29.G41 base pair in the anticodon stem. Thus, the two TilS enzymes discriminate tRNA(Ile)(2) from tRNA(Met) by strategies similar to that used by IleRS, but in distinct manners. PMID- 15894618 TI - Male genital size reflects a tradeoff between attracting mates and avoiding predators in two live-bearing fish species. AB - Male genitalia may experience more rapid, divergent evolution than any other animal character, but why? Research during the past several decades has culminated in the view that genital diversification primarily results from postmating sexual selection (e.g., sperm competition or cryptic female choice). However, the potential roles of premating sexual selection (e.g., mate choice) and natural selection have received little attention. We examined the possible importance of these mechanisms by investigating divergence in male genitalia among populations differing in predator regime for two species of live-bearing fish (Gambusia affinis in Texas and Gambusia hubbsi in The Bahamas). When controlled for body size, males exhibited a larger gonopodium (sperm-transfer organ) in predator-free environments than in predatory environments, a trend that persisted across space (multiple populations), time (multiple years), and species. By conducting laboratory experiments with G. affinis, we found that premating sexual selection seems to favor larger male genitalia (females exhibited mating preference for males having larger gonopodia), but natural selection in the presence of predatory fishes seems to favor reduced genital size (larger gonopodium size was associated with reduced burst-swimming performance, an important antipredator behavior). Although postmating sexual selection is widely presumed to be the most important mechanism driving genital diversification, these findings suggest that alternative mechanisms, particularly for organisms that cannot retract their genitalia, may also prove important. PMID- 15894619 TI - A single amino acid converts a repressor to an activator of flowering. AB - Homologous proteins occurring through gene duplication may give rise to novel functions through mutations affecting protein sequence or expression. Comparison of such homologues allows insight into how morphological traits evolve. However, it is often unclear which changes are key to determining new functions. To address these ideas, we have studied a system where two homologues have evolved clear and opposite functions in controlling a major developmental switch. In plants, flowering is a major developmental transition that is critical to reproductive success. Arabidopsis phosphatidylethanolamine-binding protein homologues TERMINAL FLOWER 1 (TFL1) and FLOWERING LOCUS T (FT) are key controllers of flowering, determining when and where flowers are made, but as opposing functions: TFL1 is a repressor, FT is an activator. We have uncovered a striking molecular basis for how these homologous proteins have diverged. Although <60% identical, we have shown that swapping a single amino acid is sufficient to convert TFL1 to FT function and vice versa. Therefore, these key residues may have strongly contributed to the selection of these important functions over plant evolution. Further, our results suggest that TFL1 and FT are highly conserved in biochemical function and that they act as repressors or activators of flowering through discrimination of structurally related interactors by a single residue. PMID- 15894620 TI - The Arabidopsis SUMO E3 ligase SIZ1 controls phosphate deficiency responses. AB - Plants sense phosphate (Pi) deficiency and initiate signaling that controls adaptive responses necessary for Pi acquisition. Herein, evidence establishes that AtSIZ1 is a plant small ubiquitin-like modifier (SUMO) E3 ligase and is a focal controller of Pi starvation-dependent responses. T-DNA insertional mutated alleles of AtSIZ1 (At5g60410) cause Arabidopsis to exhibit exaggerated prototypical Pi starvation responses, including cessation of primary root growth, extensive lateral root and root hair development, increase in root/shoot mass ratio, and greater anthocyanin accumulation, even though intracellular Pi levels in siz1 plants were similar to wild type. AtSIZ1 has SUMO E3 ligase activity in vitro, and immunoblot analysis revealed that the protein sumoylation profile is impaired in siz1 plants. AtSIZ1-GFP was localized to nuclear foci. Steadystate transcript abundances of Pi starvation-responsive genes AtPT2, AtPS2, and AtPS3 were moderate but clearly greater in siz1 seedlings than in wild type, where Pi is sufficient. Pi starvation induced the expression of these genes to the same extent in siz1 and wild-type seedlings. However, two other Pi starvation responsive genes, AtIPS1 and AtRNS1, are induced more slowly in siz1 seedlings by Pi limitation. PHR1, a MYB transcriptional activator of AtIPS1 and AtRNS1, is an AtSIZ1 sumoylation target. These results indicate that AtSIZ1 is a SUMO E3 ligase and that sumoylation is a control mechanism that acts both negatively and positively on different Pi deficiency responses. PMID- 15894621 TI - Diacylglycerol kinase iota regulates Ras guanyl-releasing protein 3 and inhibits Rap1 signaling. AB - To study the physiological function of diacylglycerol (DAG) kinase iota (DGKiota), which converts DAG to phosphatidic acid, we deleted this gene in mice. In contrast to previous studies showing that DGK isoforms decrease Ras activity, signaling downstream of Ras in embryonic fibroblasts was significantly reduced in cells lacking DGKiota. DGKs regulate Ras signaling by attenuating the function of the DAG-dependent Ras guanyl nucleotide-releasing proteins (RasGRPs). We tested whether DGKiota inhibited the four known RasGRPs and found that it inhibited only RasGRP3. In addition to activating Ras, RasGRP3 also activates Rap1, which in some cases can antagonize the function of Ras. We demonstrate that DGKiota bound to RasGRP3 and inhibited its activation of Rap1 by metabolizing DAG. This inhibition consequently affected Ras signaling. We tested the physiological consequence of deleting DGKiota by crossing wild-type or DGKiota-deficient mice with mice carrying a v-Ha-Ras transgene, and then we assessed tumor formation. We observed significantly fewer tumors in DGKiota-deficient mice. Because Rap1 can antagonize the function of Ras, our data are consistent with a model in which DGKiota regulates RasGRP3 with a predominant effect on Rap1 activity. Additionally, we found that DGKzeta, which is structurally similar to DGKiota, inhibited RasGRPs 1, 3, and 4 and predominantly affected Ras signaling. Thus, type IV DGKs regulate RasGRPs, but the downstream effects differ depending on the DGK. PMID- 15894622 TI - An ancestral oomycete locus contains late blight avirulence gene Avr3a, encoding a protein that is recognized in the host cytoplasm. AB - The oomycete Phytophthora infestans causes late blight, the potato disease that precipitated the Irish famines in 1846 and 1847. It represents a reemerging threat to potato production and is one of >70 species that are arguably the most devastating pathogens of dicotyledonous plants. Nevertheless, little is known about the molecular bases of pathogenicity in these algae-like organisms or of avirulence molecules that are perceived by host defenses. Disease resistance alleles, products of which recognize corresponding avirulence molecules in the pathogen, have been introgressed into the cultivated potato from a wild species, Solanum demissum, and R1 and R3a have been identified. We used association genetics to identify Avr3a and show that it encodes a protein that is recognized in the host cytoplasm, where it triggers R3a-dependent cell death. Avr3a resides in a region of the P. infestans genome that is colinear with the locus containing avirulence gene ATR1(NdWsB) in Hyaloperonospora parasitica, an oomycete pathogen of Arabidopsis. Remarkably, distances between conserved genes in these avirulence loci were often similar, despite intervening genomic variation. We suggest that Avr3a has undergone gene duplication and that an allele evading recognition by R3a arose under positive selection. PMID- 15894623 TI - Virulence and competitive ability in genetically diverse malaria infections. AB - Explaining parasite virulence is a great challenge for evolutionary biology. Intuitively, parasites that depend on their hosts for their survival should be benign to their hosts, yet many parasites cause harm. One explanation for this is that within-host competition favors virulence, with more virulent strains having a competitive advantage in genetically diverse infections. This idea, which is well supported in theory, remains untested empirically. Here we provide evidence that within-host competition does indeed select for high parasite virulence. We examine the rodent malaria Plasmodium chabaudi in laboratory mice, a parasite host system in which virulence can be easily monitored and competing strains quantified by using strain-specific real-time PCR. As predicted, we found a strong relationship between parasite virulence and competitive ability, so that more virulent strains have a competitive advantage in mixed-strain infections. In transmission experiments, we found that the strain composition of the parasite populations in mosquitoes was directly correlated with the composition of the blood-stage parasite population. Thus, the outcome of within-host competition determined relative transmission success. Our results imply that within-host competition is a major factor driving the evolution of virulence and can explain why many parasites harm their hosts. PMID- 15894624 TI - Molecular analysis reveals tighter social regulation of immigration in patrilocal populations than in matrilocal populations. AB - Human social organization can deeply affect levels of genetic diversity. This fact implies that genetic information can be used to study social structures, which is the basis of ethnogenetics. Recently, methods have been developed to extract this information from genetic data gathered from subdivided populations that have gone through recent spatial expansions, which is typical of most human populations. Here, we perform a Bayesian analysis of mitochondrial and Y chromosome diversity in three matrilocal and three patrilocal groups from northern Thailand to infer the number of males and females arriving in these populations each generation and to estimate the age of their range expansion. We find that the number of male immigrants is 8 times smaller in patrilocal populations than in matrilocal populations, whereas women move 2.5 times more in patrilocal populations than in matrilocal populations. In addition to providing genetic quantification of sex-specific dispersal rates in human populations, we show that although men and women are exchanged at a similar rate between matrilocal populations, there are far fewer men than women moving into patrilocal populations. This finding is compatible with the hypothesis that men are strictly controlling male immigration and promoting female immigration in patrilocal populations and that immigration is much less regulated in matrilocal populations. PMID- 15894625 TI - Dissecting coherent vibrational spectra of small proteins into secondary structural elements by sensitivity analysis. AB - The response of proteins to sequences of femtosecond infrared pulses provides a multidimensional view into their equilibrium distribution of structures and snapshot pictures of fast-triggered dynamical events. Analyzing these experiments requires advanced computational tools for assigning regions in the resulting multi-dimensional correlation plots to specific secondary-structure elements and their couplings. A differential sensitivity analysis technique based on a perturbation of the local (real space) Hamiltonian is developed to achieve that goal. Application to the amide I region of a small globular protein reveals regions associated with the alpha-helix, beta-sheet, and their coupling. Comparison of signals generated in different directions shows that the double quantum-coherence signal has a higher sensitivity to the couplings compared with the single-quantum-coherence (photon echo) technique. PMID- 15894626 TI - Subsecond reorganization of the actin network in cell motility and chemotaxis. AB - Actin networks are continuously reorganized in cells that rapidly change their shape. Applying total internal reflection fluorescence microscopy at acquisition rates of 10-20 Hz, we measured an average growth rate of 3 microm.sec(-1) for filamentous actin structures throughout the entire substrate-attached cortex of Dictyostelium cells. New filaments often proceed along preexisting ones, resulting in bundle formation concurrent with filament growth. In cells that orientate in a gradient of chemoattractant, prominent assemblies of actin enriched in the Arp2/3 complex are inserted into the network, primarily at the base of filopods that point into the direction of the gradient. We propose that high turnover rates of actin filaments confer the plasticity to the cell cortex that is required for rapid accommodation to external stimuli. PMID- 15894628 TI - Using fingerprint image quality to improve the identification performance of the U.S. Visitor and Immigrant Status Indicator Technology Program. AB - Motivated by the difficulty of biometric systems to correctly match fingerprints with poor image quality, we formulate and solve a game-theoretic formulation of the identification problem in two settings: U.S. visa applicants are checked against a list of visa holders to detect visa fraud, and visitors entering the U.S. are checked against a watchlist of criminals and suspected terrorists. For three types of biometric strategies, we solve the game in which the U.S. Government chooses the strategy's optimal parameter values to maximize the detection probability subject to a constraint on the mean biometric processing time per legal visitor, and then the terrorist chooses the image quality to minimize the detection probability. At current inspector staffing levels at ports of entry, our model predicts that a quality-dependent two-finger strategy achieves a detection probability of 0.733, compared to 0.526 under the quality independent two-finger strategy that is currently implemented at the U.S. border. Increasing the staffing level of inspectors offers only minor increases in the detection probability for these two strategies. Using more than two fingers to match visitors with poor image quality allows a detection probability of 0.949 under current staffing levels, but may require major changes to the current U.S. biometric program. The detection probabilities during visa application are approximately 11-22% smaller than at ports of entry for all three strategies, but the same qualitative conclusions hold. PMID- 15894627 TI - Sexually dimorphic expression of trkB, a Z-linked gene, in early posthatch zebra finch brain. AB - Sexual differentiation of the zebra finch (Taeniopygia guttata) neural song circuit is thought to be initiated by sex differences in sex chromosome gene expression in brain cells. One theory is that Z-linked genes, present in the male's ZZ genome at double the dose of females' (ZW), are expressed at higher levels and trigger masculine patterns of development. We report here that trkB (tyrosine kinase receptor B) is Z-linked in zebra finches. trkB is the receptor for neurotrophic factors BDNF and neurotrophin 4, and mediates their influence on neuronal survival, migration, and specification. trkB mRNA is expressed at a higher level in the male telencephalon or whole brain than in corresponding regions of the female in adulthood, and at posthatch day (P) 6, when the song circuit is undergoing sexual differentiation. Moreover, this expression is higher in the song nucleus high vocal center (HVC) than in the surrounding telencephalon at P6, and in males relative to females. In addition, trkB protein is expressed more highly in male than female whole brain at P6. These results establish trkB as a candidate factor that contributes to masculine differentiation of HVC because of its Z-linkage, which leads to sex differences in expression. BDNF is known to be stimulated by estrogen and to be expressed at higher levels in males than females at later ages in HVC. Thus, the trkB-BDNF system may be a focal point for convergent masculinizing influences of Z-linked factors and hormones. PMID- 15894629 TI - Helix packing and orientation in the transmembrane dimer of gp55-P of the spleen focus forming virus. AB - gp55-P is a dimeric membrane protein with a single transmembrane helix that is coded by the env gene of the polycythemic strain of the spleen focus forming virus. gp55-P activates the erythropoietin (Epo) receptor through specific transmembrane helix interactions, leading to Epo-independent growth of erythroid progenitors and eventually promoting erythroleukemia. We describe the use of magic angle spinning deuterium NMR to establish the structure of the transmembrane dimer of gp55-P in model membranes. Comparison of the deuterium lineshapes of leucines in the center (Leu(396-399)) and at the ends (Leu(385), Leu(407)) of the transmembrane sequence shows that gp55-P has a right-handed crossing angle with Leu(399) packed in the dimer interface. We discuss the implications of the structure of the gp55-P transmembrane dimer for activation of the Epo receptor. PMID- 15894630 TI - Interfacial water as a "hydration fingerprint" in the noncognate complex of BamHI. AB - The molecular code of specific DNA recognition by proteins as a paradigm in molecular biology remains an unsolved puzzle primarily because of the subtle interplay between direct protein-DNA interaction and the indirect contribution from water and ions. Transformation of the nonspecific, low affinity complex to a specific, high affinity complex is accompanied by the release of interfacial water molecules. To provide insight into the conversion from the loose to the tight form, we characterized the structure and energetics of water at the protein DNA interface of the BamHI complex with a noncognate sequence and in the specific complex. The fully hydrated models were produced with Grand Canonical Monte Carlo simulations. Proximity analysis shows that water distributions exhibit sequence dependent variations in both complexes and, in particular, in the noncognate complex they discriminate between the correct and the star site. Variations in water distributions control the number of water molecules released from a given sequence upon transformation from the loose to the tight complex as well as the local entropy contribution to the binding free energy. We propose that interfacial waters can serve as a "hydration fingerprint" of a given DNA sequence. PMID- 15894631 TI - Orientation of the myosin light chain region by single molecule total internal reflection fluorescence polarization microscopy. AB - To study the orientation and dynamics of myosin, we measured fluorescence polarization of single molecules and ensembles of myosin decorating actin filaments. Engineered chicken gizzard regulatory light chain (RLC), labeled with bisiodoacetamidorhodamine at cysteine residues 100 and 108 or 104 and 115, was exchanged for endogenous RLC in rabbit skeletal muscle HMM or S1. AEDANS-labeled actin, fully decorated with labeled myosin fragment or a ratio of approximately 1:1000 labeled:unlabeled myosin fragment, was adhered to a quartz slide. Eight polarized fluorescence intensities were combined with the actin orientation from the AEDANS fluorescence to determine the axial angle (relative to actin), the azimuthal angle (around actin), and RLC mobility on the <<10 ms timescale. Order parameters of the orientation distributions from heavily labeled filaments agree well with comparable measurements in muscle fibers, verifying the technique. Experiments with HMM provide sufficient angular resolution to detect two orientations corresponding to the two heads in rigor. Experiments with S1 show a single orientation intermediate to the two seen for HMM. The angles measured for HMM are consistent with heads bound on adjacent actin monomers of a filament, under strain, similar to predictions based on ensemble measurements made on muscle fibers with electron microscopy and spectroscopic experiments. PMID- 15894632 TI - Measurement of single macromolecule orientation by total internal reflection fluorescence polarization microscopy. AB - A new approach is presented for measuring the three-dimensional orientation of individual macromolecules using single molecule fluorescence polarization (SMFP) microscopy. The technique uses the unique polarizations of evanescent waves generated by total internal reflection to excite the dipole moment of individual fluorophores. To evaluate the new SMFP technique, single molecule orientation measurements from sparsely labeled F-actin are compared to ensemble-averaged orientation data from similarly prepared densely labeled F-actin. Standard deviations of the SMFP measurements taken at 40 ms time intervals indicate that the uncertainty for individual measurements of axial and azimuthal angles is approximately 10 degrees at 40 ms time resolution. Comparison with ensemble data shows there are no substantial systematic errors associated with the single molecule measurements. In addition to evaluating the technique, the data also provide a new measurement of the torsional rigidity of F-actin. These measurements support the smaller of two values of the torsional rigidity of F actin previously reported. PMID- 15894633 TI - Quaternary structures of intermediately ligated human hemoglobin a and influences from strong allosteric effectors: resonance Raman investigation. AB - The Fe-histidine stretching (nu(Fe-His)) frequency was determined for deoxy subunits of intermediately ligated human hemoglobin A in equilibrium and CO photodissociated picosecond transient species in the presence and absence of strong allosteric effectors like inositol(hexakis)phosphate, bezafibrate, and 2,3 bisphosphoglycerate. The nu(Fe-His) frequency of deoxyHb A was unaltered by the effectors. The T-to-R transition occurred around m = 2-3 in the absence of effectors but m > 3.5 in their presence, where m is the average number of ligands bound to Hb and was determined from the intensity of the nu(4) band measured in the same experiment. The alpha1-beta2 subunit contacts revealed by ultraviolet resonance Raman spectra, which were distinctly different between the T and R states, remained unchanged by the effectors. This observation would solve the recent discrepancy that the strong effectors remove the cooperativity of oxygen binding in the low-affinity limit, whereas the (1)H NMR spectrum of fully ligated form exhibits the pattern of the R state. PMID- 15894634 TI - Membrane elasticity in giant vesicles with fluid phase coexistence. AB - Biological membranes are known to contain compositional heterogeneities, often termed rafts, with distinguishable composition and function, and these heterogeneities participate in vigorous transport processes. Membrane lipid phase coexistence is expected to modulate these processes through the differing mechanical properties of the bulk domains and line tension at phase boundaries. In this contribution, we compare the predictions from a shape theory derived for vesicles with fluid phase coexistence to the geometry of giant unilamellar vesicles with coexisting liquid-disordered (L(d)) and liquid-ordered (L(o)) phases. We find a bending modulus for the L(o) phase higher than that of the L(d) phase and a saddle-splay (Gauss) modulus difference with the Gauss modulus of the L(o) phase being more negative than the L(d) phase. The Gauss modulus critically influences membrane processes that change topology, such as vesicle fission or fusion, and could therefore be of significant biological relevance in heterogeneous membranes. Our observations of experimental vesicle geometries being modulated by Gaussian curvature moduli differences confirm the prediction by the theory of Juelicher and Lipowsky. PMID- 15894635 TI - Two-photon absorption of bacteriorhodopsin: formation of a red-shifted thermally stable photoproduct F620. AB - By means of high-intensity 532 nm laser pulses, a photochemical conversion of the initial B(570) state of bacteriorhodopsin (BR) to a stable photoproduct absorbing maximally at approximately 620 nm in BR suspensions and at approximately 610 nm in BR films is induced. This state, which we named F(620), is photochemically further converted to a group of three products with maximal absorptions in the wavelength range from 340 nm to 380 nm, which show identical spectral properties to the so-called P(360) state reported in the literature. The photoconversion from B(570) to F(620) is most likely a resonant two-photon absorption induced step. The formation of F(620) and P(360) leads to a distinguished photo-induced permanent optical anisotropy in BR films. The spectral dependence of the photo induced anisotropy and the anisotropy orientations at the educt (B(570)) and product (F(620)) wavelengths are strong indicators that F(620) is formed in a direct photochemical step from B(570). The chemical nature of the P(360) products probably is that of a retro-retinal containing BR, but the structural characteristics of the F(620) state are still unclear. The photo-induced permanent anisotropy induced by short laser pulses in BR films helps to better understand the photochemical pathways related to this transition, and it is interesting in view of potential applications as this feature is the molecular basis for permanent optical data storage using BR films. PMID- 15894636 TI - Backbone dynamics of a symmetric calmodulin dimer in complex with the calmodulin binding domain of the basic-helix-loop-helix transcription factor SEF2-1/E2-2: a highly dynamic complex. AB - Calmodulin (CaM) interacts specifically as a dimer with some dimeric basic-Helix Loop-Helix (bHLH) transcription factors via a novel high affinity binding mode. Here we report a study of the backbone dynamics by (15)N-spin relaxation on the CaM dimer in complex with a dimeric peptide that mimics the CaM binding region of the bHLH transcription factor SEF2-1. The relaxation data were measured at multiple magnetic fields, and analyzed in a model-free manner using in-house written software designed to detect nanosecond internal motion. Besides picosecond motions, all residues also experience internal motion with an effective correlation time of approximately 2.5 ns with squared order parameter (S(2)) of approximately 0.75. Hydrodynamic calculations suggest that this can be attributed to motions of the N- and C-terminal domains of the CaM dimer in the complex. Moreover, residues with significant exchange broadening are found. They are clustered in the CaM:SEF2-1mp binding interface, the CaM:CaM dimer interface, and in the flexible helix connecting the CaM N- and C-terminal domains, and have similar exchange times (approximately 50 micros), suggesting a cooperative mechanism probably caused by protein:protein interactions. The dynamic features presented here support the conclusion that the conformationally heterogeneous bHLH mimicking peptide trapped inside the CaM dimer exchanges between different binding sites on both nanosecond and microsecond timescales. Nature has thus found a way to specifically recognize a relatively ill-fitting target. This novel mode of target-specific binding, which neither belongs to lock-and-key nor induced-fit binding, is characterized by dimerization and continuous exchange between multiple flexible binding alternatives. PMID- 15894637 TI - Determining diffusion coefficients in inhomogeneous tissues using fluorescence recovery after photobleaching. AB - Diffusion plays an important role in the transport of nutrients and signaling molecules in cartilaginous tissues. Diffusion coefficients can be measured by fluorescence recovery after photobleaching (FRAP). Available methods to analyze FRAP data, however, assume homogeneity in the environment of the bleached area and neglect geometrical restrictions to diffusion. Hence, diffusion coefficients in inhomogeneous materials, such as most biological tissues, cannot be assessed accurately. In this study, a new method for analyzing data from FRAP measurements has been developed, which is applicable to inhomogeneous tissues. It is based on a fitting procedure of the intensity recovery after photobleaching with a two dimensional finite element analysis, which includes Fick's law for diffusion. The finite element analysis can account for distinctive diffusivity in predefined zones, which allows determining diffusion coefficients in inhomogeneous samples. The method is validated theoretically and experimentally in both homogeneous and inhomogeneous tissues and subsequently applied to the proliferation zone of the growth plate. Finally, the importance of accounting for inhomogeneities, for appropriate assessment of diffusivity in inhomogeneous tissues, is illustrated. PMID- 15894638 TI - Structural changes of actin-bound myosin heads after a quick length change in frog skeletal muscle. AB - Changes in the x-ray diffraction pattern from a frog skeletal muscle were recorded after a quick release or stretch, which was completed within one millisecond, at a time resolution of 0.53 ms using the high-flux beamline at the SPring-8 third-generation synchrotron radiation facility. Reversibility of the effects of the length changes was checked by quickly restoring the muscle length. Intensities of seven reflections were measured. A large, instantaneous intensity drop of a layer line at an axial spacing of 1/10.3 nm(-1) after a quick release and stretch, and its partial recovery by reversal of the length change, indicate a conformational change of myosin heads that are attached to actin. Intensity changes on the 14.5-nm myosin layer line suggest that the attached heads alter their radial mass distribution upon filament sliding. Intensity changes of the myosin reflections at 1/21.5 and 1/7.2 nm(-1) are not readily explained by a simple axial swing of cross-bridges. Intensity changes of the actin-based layer lines at 1/36 and 1/5.9 nm(-1) are not explained by it either, suggesting a structural change in actin molecules. PMID- 15894639 TI - Molecular dynamics of a protein surface: ion-residues interactions. AB - Time-resolved measurements indicated that protons could propagate on the surface of a protein or a membrane by a special mechanism that enhanced the shuttle of the proton toward a specific site. It was proposed that a suitable location of residues on the surface contributes to the proton shuttling function. In this study, this notion was further investigated by the use of molecular dynamics simulations, where Na(+) and Cl(-) are the ions under study, thus avoiding the necessity for quantum mechanical calculations. Molecular dynamics simulations were carried out using as a model a few Na(+) and Cl(-) ions enclosed in a fully hydrated simulation box with a small globular protein (the S6 of the bacterial ribosome). Three independent 10-ns-long simulations indicated that the ions and the protein's surface were in equilibrium, with rapid passage of the ions between the protein's surface and the bulk. However, it was noted that close to some domains the ions extended their duration near the surface, thus suggesting that the local electrostatic potential hindered their diffusion to the bulk. During the time frame in which the ions were detained next to the surface, they could rapidly shuttle between various attractor sites located under the electrostatic umbrella. Statistical analysis of the molecular dynamics and electrostatic potential/entropy consideration indicated that the detainment state is an energetic compromise between attractive forces and entropy of dilution. The similarity between the motion of free ions next to a protein and the proton transfer on the protein's surface are discussed. PMID- 15894640 TI - Enzyme activity and flexibility at very low hydration. AB - Recent measurements have demonstrated enzyme activity at hydrations as low as 3%. This raises the question of whether hydration-induced enzyme flexibility is important for activity. Here, to address this, picosecond dynamic neutron scattering experiments are performed on pig liver esterase powders at 0%, 3%, 12%, and 50% hydration by weight and at temperatures ranging from 120 to 300 K. At all temperatures and hydrations, significant quasielastic scattering intensity is found in the protein, indicating the presence of anharmonic, diffusive motion. As the hydration increases, a temperature-dependent dynamical transition appears and strengthens involving additional diffusive motion. The implication of these results is that, although the additional hydration-induced diffusive motion in the protein detected here may be related to increased activity, it is not required for the enzyme to function. PMID- 15894641 TI - Changes of enzyme activity in lipid signaling pathways related to substrate reordering. AB - The static fluid mosaic model of biological membranes has been progressively complemented by a dynamic membrane model that includes phospholipid reordering in domains that are proposed to extend from nanometers to microns. Kinetic models for lipolytic enzymes have only been developed for homogeneous lipid phases. In this work, we develop a generalization of the well-known surface dilution kinetic theory to cases where, in a same lipid phase, both domain and nondomain phases coexist. Our model also allows understanding the changes in enzymatic activity due to a decrease of free substrate concentration when domains are induced by peptides. This lipid reordering and domain dynamics can affect the activity of lipolytic enzymes, and can provide a simple explanation for how basic peptides, with a strong direct interaction with acidic phospholipids (such as beta-amyloid peptide), may cause a complex modulation of the activities of many important enzymes in lipid signaling pathways. PMID- 15894642 TI - Osmolyte trimethylamine-N-oxide does not affect the strength of hydrophobic interactions: origin of osmolyte compatibility. AB - Osmolytes are small organic solutes accumulated at high concentrations by cells/tissues in response to osmotic stress. Osmolytes increase thermodynamic stability of folded proteins and provide protection against denaturing stresses. The mechanism of osmolyte compatibility and osmolyte-induced stability has, therefore, attracted considerable attention in recent years. However, to our knowledge, no quantitative study of osmolyte effects on the strength of hydrophobic interactions has been reported. Here, we present a detailed molecular dynamics simulation study of the effect of the osmolyte trimethylamine-N-oxide (TMAO) on hydrophobic phenomena at molecular and nanoscopic length scales. Specifically, we investigate the effects of TMAO on the thermodynamics of hydrophobic hydration and interactions of small solutes as well as on the folding unfolding conformational equilibrium of a hydrophobic polymer in water. The major conclusion of our study is that TMAO has almost no effect either on the thermodynamics of hydration of small nonpolar solutes or on the hydrophobic interactions at the pair and many-body level. We propose that this neutrality of TMAO toward hydrophobic interactions-one of the primary driving forces in protein folding-is at least partially responsible for making TMAO a "compatible" osmolyte. That is, TMAO can be tolerated at high concentrations in organisms without affecting nonspecific hydrophobic effects. Our study implies that protein stabilization by TMAO occurs through other mechanisms, such as unfavorable water mediated interaction of TMAO with the protein backbone, as suggested by recent experimental studies. We complement the above calculations with analysis of TMAO hydration and changes in water structure in the presence of TMAO molecules. TMAO is an amphiphilic molecule containing both hydrophobic and hydrophilic parts. The precise balance of the effects of hydrophobic and hydrophilic segments of the molecule appears to explain the virtual noneffect of TMAO on the strength of hydrophobic interactions. PMID- 15894643 TI - Surface viscosity, diffusion, and intermonolayer friction: simulating sheared amphiphilic bilayers. AB - The flow properties of an amphiphilic bilayer are studied in molecular dynamics simulations, by exposing a coarse grained model bilayer to two shear flows directed along the bilayer surface. The first field, with a vorticity perpendicular to the bilayer, induces a regular shear deformation, allowing a direct calculation of the surface viscosity. In experiments this property is measured indirectly, by relating it to the diffusion coefficient of a tracer particle through the Saffman-Einstein expression. The current calculations provide an independent test of this relation. The second flow field, with a vorticity parallel to the bilayer, causes the two constituent monolayers to slide past one another, yielding the interlayer friction coefficient. PMID- 15894644 TI - Assessing oligomerization of membrane proteins by four-pulse DEER: pH-dependent dimerization of NhaA Na+/H+ antiporter of E. coli. AB - The pH dependence of the structure of the main Na(+)/H(+) antiporter NhaA of Escherichia coli is studied by continuous-wave (CW) and pulse electron paramagnetic resonance (EPR) techniques on singly spin-labeled mutants. Residues 225 and 254 were selected for site-directed spin labeling, as previous work suggested that they are situated in domains undergoing pH-dependent structural changes. A well-defined distance of 4.4 nm between residues H225R1 in neighboring molecules is detected by a modulation in double electron-electron resonance data. This indicates that NhaA exists as a dimer, as previously suggested by a low resolution electron density map and cross-linking experiments. The modulation depth decreases reversibly when pH is decreased from 8 to 5.8. A quantitative analysis suggests a dimerization equilibrium, which depends moderately on pH. Furthermore, the mobility and polarity of the environment of a spin label attached to residue 225 change only slightly with changing pH, while no other changes are detected by CW EPR. As antiporter activity of NhaA changes drastically in the studied pH range, residues 225 and 254 are probably located not in the sensor or ion translocation sites themselves but in domains that convey the signal from the pH sensor to the translocation site. PMID- 15894645 TI - Position-sensitive scanning fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) uses a stationary laser beam to illuminate a small sample volume and analyze the temporal behavior of the fluorescence fluctuations within the stationary observation volume. In contrast, scanning FCS (SFCS) collects the fluorescence signal from a moving observation volume by scanning the laser beam. The fluctuations now contain both temporal and spatial information about the sample. To access the spatial information we synchronize scanning and data acquisition. Synchronization allows us to evaluate correlations for every position along the scanned trajectory. We use a circular scan trajectory in this study. Because the scan radius is constant, the phase angle is sufficient to characterize the position of the beam. We introduce position-sensitive SFCS (PSFCS), where correlations are calculated as a function of lag time and phase. We present the theory of PSFCS and derive expressions for diffusion, diffusion in the presence of flow, and for immobilization. To test PSFCS we compare experimental data with theory. We determine the direction and speed of a flowing dye solution and the position of an immobilized particle. To demonstrate the feasibility of the technique for applications in living cells we present data of enhanced green fluorescent protein measured in the nucleus of COS cells. PMID- 15894646 TI - Role of intramolecular vibrations in long-range electron transfer between pheophytin and ubiquinone in bacterial photosynthetic reaction centers. AB - The dynamics of the elementary electron transfer step between pheophytin and primary ubiquinone in bacterial photosynthetic reaction centers is investigated by using a discrete state approach, including only the intramolecular normal modes of vibration of the two redox partners. The whole set of normal coordinates of the acceptor and donor groups have been employed in the computations of the Hamiltonian matrix, to reliably account both for shifts and mixing of the normal coordinates, and for changes in vibrational frequencies upon ET. It is shown that intramolecular modes provide not only a discrete set of states more strongly coupled to the initial state but also a quasicontinuum of weakly coupled states, which account for the spreading of the wave packet after ET. The computed transition probabilities are sufficiently high for asserting that electron transfer from bacteriopheophytin to the primary quinone can occur via tunneling solely promoted by intramolecular modes; the transition times, computed for different values of the electronic energy difference and coupling term, are of the same order of magnitude (10(2) ps) of the observed one. PMID- 15894647 TI - Differential osmotic behavior of water components in living skeletal muscle resolved by 1H-NMR. AB - Using frog sartorius muscle, we observed transverse relaxation processes of (1)H NMR signals from myowater. The process could be well described by four characteristic exponentials: the extremely slow exponential of relaxation time constant T(2) > 0.4 s, the slow one of T(2) approximately 0.15 s, the intermediate one of 0.03 s < T(2) < 0.06 s, and the rapid one of T(2) < 0.03 s. Addition of isotonic extracellular solution affected only the extremely slow exponential, linearly increasing its amplitude and gradually increasing its T(2) toward that of the bulk solution (1.7 s). Therefore, this exponential should represent extracellular surplus solution independently of the other exponentials. At two thirds to three times the isotonicity, the amplitude of the intermediate exponential showed normal osmotic behavior in parallel with the volume change of the myofilament lattice measured with x-ray diffraction. In the same tonicity range, the amplitude of the rapid exponential showed converse osmotic behavior. Lower tonicities increased the amplitude of only the slow exponential. Studied tonicities did not affect the T(2) values. The distinct osmotic behavior indicated that each characteristic exponential could be viewed as a distinct water group. In addition, the converse osmotic behavior suggested that the rapid exponential would not be a static water layer on the macromolecule surface. PMID- 15894648 TI - Dynamics of vesicles in a wall-bounded shear flow. AB - We report a detailed study of the behavior (shapes, experienced forces, velocities) of giant lipid vesicles subjected to a shear flow close to a wall. Vesicle buoyancy, size, and reduced volume were separately varied. We show that vesicles are deformed by the flow and exhibit a tank-treading motion with steady orientation. Their shapes are characterized by two nondimensional parameters: the reduced volume and the ratio of the shear stress with the hydrostatic pressure. We confirm the existence of a force, able to lift away nonspherical buoyant vesicles from the substrate. We give the functional variation and the value of this lift force (up to 150 pN in our experimental conditions) as a function of the relevant physical parameters: vesicle-substrate distance, wall shear rate, viscosity of the solution, vesicle size, and reduced volume. Circulating deformable cells disclosing a nonspherical shape also experience this force of viscous origin, which contributes to take them away from the endothelium and should be taken into account in studies on cell adhesion in flow chambers, where cells membrane and the adhesive substrate are in relative motion. Finally, the kinematics of vesicles along the flow direction can be described in a first approximation with a model of rigid spheres. PMID- 15894649 TI - The predominance of the environment over genes in cancer causation: implications for genetic epidemiology. PMID- 15894650 TI - Analyzing patterns of staining in immunohistochemical studies: application to a study of prostate cancer recurrence. AB - BACKGROUND: Immunohistochemical studies use antibodies to stain tissues with the goal of quantifying protein expression. However, protein expression is often heterogeneous resulting in variable degrees and patterns of staining. This problem is particularly acute in prostate cancer, where tumors are infiltrative and heterogeneous in nature. In this article, we introduce analytic approaches that explicitly consider both the frequency and intensity of tissue staining. METHODS: Compositional data analysis is a technique used to analyze vectors of unit-sum proportions, such as those obtained from soil sample studies or species abundance surveys. We summarized specimen staining patterns by the proportion of cells staining at mild, moderate, and intense levels and used compositional data analysis to summarize and compare the resulting staining profiles. RESULTS: In a study of Syndecan-1 staining patterns among 44 localized prostate cancer cases with Gleason score 7 disease, compositional data analysis did not detect a statistically significant difference between the staining patterns in recurrent (n = 22) versus nonrecurrent (n = 22) patients. Results indicated only modest increases in the proportion of cells staining at a moderate intensity in the recurrent group. In contrast, an analysis that compared quantitative scores across groups indicated a (borderline) significant increase in staining in the recurrent group (P = 0.05, t test). CONCLUSIONS: Compositional data analysis offers a novel analytic approach for immunohistochemical studies, providing greater insight into differences in staining patterns between groups, but possibly lower statistical power than existing, score-based methods. When appropriate, we recommend conducting a compositional data analysis in addition to a standard score-based analysis. PMID- 15894651 TI - Sex hormones, risk factors, and risk of estrogen receptor-positive breast cancer in older women: a long-term prospective study. AB - OBJECTIVE: Antiestrogens reduce the risk of estrogen receptor-positive (ER+) but not ER-negative (ER-) breast cancer. Women at high risk of ER+ cancer would be the most likely to benefit from these treatments, but the best approach to predicting ER+ cancer is uncertain. METHODS: We prospectively assessed putative risk factors for breast cancer and archived serum at -190 degrees C from a community-based cohort of 7,676 women ages > or =65 years who had no history of breast cancer. Follow-up for breast cancer over 10.5 years was 99% complete. Using a case-cohort design, we measured baseline levels of estradiol and testosterone in 196 cases of invasive ER+ cancer and 378 randomly selected controls. RESULTS: Women whose testosterone level in highest two quintiles had a 4-fold increased risk of ER+ breast cancer (P < 0.0001). High estradiol concentration also indicated an increased risk but was not a significant predictor after adjustment for testosterone. Women with >16 years of education had a 2.1 times increased risk (P = 0.03) of ER+ cancer, but no other risk factors were significantly related to an increased risk of ER+ cancer. Women with a family history of breast cancer had a 2.9-fold increased risk of ER- cancer (P = 0.002) but no increased risk of ER+ cancer (relative hazard = 1.2, 0.8-1.8). CONCLUSIONS: High serum testosterone and advanced education predicted ER+ breast cancer. If confirmed, high testosterone level may be more accurate than family history of breast cancer and other conventional risk factors for identifying older women who are most likely to benefit from antiestrogen chemoprevention. PMID- 15894652 TI - Mammographic features and subsequent risk of breast cancer: a comparison of qualitative and quantitative evaluations in the Guernsey prospective studies. AB - Mammographic features are known to be associated with breast cancer but the magnitude of the effect differs markedly from study to study. Methods to assess mammographic features range from subjective qualitative classifications to computer-automated quantitative measures. We used data from the UK Guernsey prospective studies to examine the relative value of these methods in predicting breast cancer risk. In all, 3,211 women ages > or =35 years who had a mammogram taken in 1986 to 1989 were followed-up to the end of October 2003, with 111 developing breast cancer during this period. Mammograms were classified using the subjective qualitative Wolfe classification and several quantitative mammographic features measured using computer-based techniques. Breast cancer risk was positively associated with high-grade Wolfe classification, percent breast density and area of dense tissue, and negatively associated with area of lucent tissue, fractal dimension, and lacunarity. Inclusion of the quantitative measures in the same model identified area of dense tissue and lacunarity as the best predictors of breast cancer, with risk increasing by 59% [95% confidence interval (95% CI), 29-94%] per SD increase in total area of dense tissue but declining by 39% (95% CI, 53-22%) per SD increase in lacunarity, after adjusting for each other and for other confounders. Comparison of models that included both the qualitative Wolfe classification and these two quantitative measures to models that included either the qualitative or the two quantitative variables showed that they all made significant contributions to prediction of breast cancer risk. These findings indicate that breast cancer risk is affected not only by the amount of mammographic density but also by the degree of heterogeneity of the parenchymal pattern and, presumably, by other features captured by the Wolfe classification. PMID- 15894653 TI - Hormone replacement therapy, percent mammographic density, and sensitivity of mammography. AB - OBJECTIVE: We examine to what extent the lower mammographic sensitivity found in hormone replacement therapy (HRT) users can be explained by any association of HRT use with higher mammographic density and more difficult to detect cancers. METHODS: We used logistic regression to estimate the odds of a false-negative screen (a breast cancer diagnosed in the 24 months after a negative screening examination) for HRT users and to estimate, and adjust for, mammographic density (measured on a continuous scale, blinded, using a reliable, computer-assisted method), tumor characteristics (size, grade, and morphology), and potential confounders (age, symptom status, family history, and prior screening) among women ages > or =55 years who attended BreastScreen Victoria for first round screening mammography in 1994 and 1995 (1,086 breast cancers) and for subsequent round screening (471 breast cancers) in 1995 and 1996. RESULTS: After adjusting for confounders, HRT users were more likely to have a false-negative screen [first round: odds ratio (OR), 1.99; 95% confidence interval (95% CI), 1.4-2.9; subsequent round: OR, 2.29; 95% CI, 1.4-3.8]. This effect was modestly attenuated by adjusting for mammographic density (first round: OR, 1.54; 95% CI, 1.0-2.3; subsequent round: OR, 1.97; 95% CI, 1.2-3.3). Adjusting for tumor characteristics resulted in a modest increase in the odds of a false negative at first round but had no effect at subsequent round. CONCLUSIONS: Mammographic density only partly explains the effect of HRT on sensitivity. Further research needs to clarify whether hyperemic breast tissue changes affect cancer detectability in HRT users as well as the possibility that the quality of mammography may be poor in some HRT users. PMID- 15894654 TI - Insulin-like growth factor-I, IGF-binding protein-3, and mammographic breast density. AB - Some studies have suggested that insulin-like growth factor (IGF) pathway is related to premenopausal breast density, one of the strongest known breast cancer risk factors. This study was designed specifically to test the hypothesis that higher levels of IGF-I and lower levels of IGF-binding protein (IGFBP)-3 are associated with high mammographic breast density among premenopausal but not among postmenopausal women. A total of 783 premenopausal and 791 postmenopausal healthy women were recruited during screening mammography examinations. Blood samples were collected at the time of mammography, and plasma IGF-I and IGFBP-3 levels were measured by ELISA. Mammographic breast density was estimated using a computer-assisted method. Spearman's partial correlation coefficients (r(s)) were used to evaluate the associations. Adjusted mean breast density was assessed by joint levels of IGF-I and IGFBP-3 using generalized linear models. Among premenopausal women, high levels of IGF-I and low levels of IGFBP-3 were independently correlated with high breast density (r(s) = 0.083; P = 0.021 and r(s) = -0.124; P = 0.0005, respectively). Correlation of IGF-I with breast density was stronger among women in the lowest tertile of IGFBP-3 than among those in the highest tertile of IGFBP-3 (r(s) = 0.138; P = 0.027 and r(s) = 0.039; P = 0.530, respectively). In contrast, the correlation of IGFBP-3 with breast density was stronger among women in the highest tertile of IGF-I than among those in the lowest tertile of IGF-I (r(s) = -0.150; P = 0.016 and r(s) = 0.008; P = 0.904, respectively). Women in the combined top tertile of IGF-I and bottom tertile of IGFBP-3 had higher mean breast density than those in the combined bottom tertile of IGF-I and top tertile of IGFBP-3 (53.8% versus 40.9%; P = 0.014). No significant association was observed among postmenopausal women. Our findings confirm that IGF-I and IGFBP-3 are associated with breast density among premenopausal women. They provide additional support for the idea that, among premenopausal women, these growth factors may affect breast cancer risk, at least in part, through their influence on breast tissue morphology as reflected on mammogram. PMID- 15894655 TI - Dietary and plasma lycopene and the risk of breast cancer. AB - Lycopene is potentially effective in the prevention of breast cancer from laboratory and observational studies. Among 39,876 women initially free of cardiovascular disease and cancer, we first conducted a prospective cohort study of dietary lycopene and its food sources. Participants completed a baseline food frequency questionnaire and provided self-reports of breast cancer risk factors. Dietary lycopene levels were divided into quintiles, and lycopene food sources were categorized. During 9.9 years of follow-up, 1,076 breast cancer cases were confirmed by medical record review. In a nested case-control study, we then identified 508 breast cancer cases and 508 controls matched by age, smoking, and follow-up time. Plasma lycopene and other carotenoids were measured. In the prospective cohort study, women with increasing quintiles of dietary lycopene had multivariate relative risks (RR) of breast cancer of 1.00 (ref), 0.95, 1.00, 1.10, and 1.00 (P, linear trend = 0.71). Women consuming <1.5, 1.5 to <4, 4 to <7, 7 to <10, and > or =10 servings/week of tomato-based products had RRs of 1.00 (ref), 1.00, 1.20, 1.18, and 1.16 (P, linear trend = 0.11). No individual lycopene food sources were associated with breast cancer. In the nested case control study, women in increasing quartiles of plasma lycopene had multivariate RRs of breast cancer of 1.00 (ref), 0.95, 1.15, and 0.93 (P, linear trend = 0.86). The stepwise addition of individual plasma carotenoids did not impact the RRs for plasma lycopene, nor were other carotenoids associated with breast cancer. In conclusion, neither higher dietary nor plasma lycopene levels were associated with a reduced risk of breast cancer in middle-aged and older women. PMID- 15894656 TI - Ductal lavage of fluid-yielding and non-fluid-yielding ducts in BRCA1 and BRCA2 mutation carriers and other women at high inherited breast cancer risk. AB - OBJECTIVE: Nipple fluid production and atypical breast duct cells in women at high risk of breast cancer have been associated with further increased risk. Most publications on ductal lavage for cell collection report cannulating fluid yielding ducts only. We report lavage of fluid-yielding and non-fluid-yielding ducts in women at high inherited breast cancer risk. METHODS: A pilot breast cancer screening study including ductal lavage was conducted in 75 women at high inherited risk, 56 (74.7%) of whom had BRCA1/2 mutations. Ductal lavage was attempted in any duct identifiable with a catheter. RESULTS: Ducts were successfully catheterized in 60 of 75 patients (80%). Successfully catheterized patients were younger (median age 41 versus 53 years, P = 0.0003) and more often premenopausal (51.7% versus 20%, P = 0.041). Thirty-one successfully catheterized patients [51.6%, 95% confidence interval (39.4-63.9%)] had non-fluid-yielding ducts only. Seventeen patients [28.3% (18.5-40.9%)] had atypical cells. Twelve of seventeen [70.6% (46.8-87.2%)] samples with atypia were from non-fluid-yielding ducts. Patients with non-fluid-yielding ducts (versus fluid-yielding ducts) were more likely to have had prior cancer (48.4% versus 17.2%, P = 0.014) or chemotherapy (45.2% versus 17.2%, P = 0.027); this was also true in patients with atypia from non-fluid-yielding ducts. CONCLUSION: Successfully lavaged women were younger and more often premenopausal. Atypical cells can be found in non-fluid yielding ducts in patients at high inherited breast cancer risk. Non-fluid yielding ducts, and atypia from non-fluid-yielding ducts, are more common in patients with prior cancer and chemotherapy. Larger studies are needed to identify risk factors and prognostic significance associated with atypia and non fluid-yielding ducts in high-risk populations, and define their role as biomarkers. PMID- 15894657 TI - Genetic polymorphisms of SULT1A1 and SULT1E1 and the risk and survival of breast cancer. AB - We examined whether common single nucleotide polymorphisms (SNP) in SULT1A1 (c.779G>A, *14A>G, and *85C>T) and SULT1E1 (IVS1-447C>A, IVS4-1653T>C, and *959G>A) genes influenced the risk and survival of breast cancer. Our study population consisted of 989 histologically confirmed sporadic breast cancer patients and 1,054 controls without history of cancer recruited from three teaching hospitals in Seoul. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated by logistic regression model. In the survival analysis for 529 breast cancer patients with completed treatments, the hazard ratios (HR) were calculated with Cox proportional hazard model. Women with the SULT1E1 *959 GA/AA genotype had a moderately decreased breast cancer risk compared with those with the GG genotypes (OR, 0.8; 95% CI, 0.70-1.00). When the haplotypes were considered, the homozygous *959 AA genotype together with the IVS4-1653 T>C base change (CTA-CCA haplotype) was associated with halved breast cancer risk (OR, 0.5; 95% CI, 0.24-0.88) compared with the wild type CTG-CTG haplotype. No other significant overall association was observed between the SULT1A1 and SULT1E1 SNPs nor haplotypes and breast cancer risk. When stratified by survival, patients with the SULT1E1 IVS4-1653 TC/CC genotypes showed a >3-fold risk of recurrence (HR, 3.2; 95% CI, 1.39-7.48) compared with those with the TT genotype. Moreover, when the haplotypes were considered, the SULT1E1 *959 G>A base change together with the IVS4-1653 T>C base change (CTG-CCA haplotype) was associated with a >4-fold risk of breast cancer (OR, 4.2; 95% CI, 1.15-15.15). These findings suggest that genetic polymorphisms of SULT1E1 are associated with increased risk and a disease free survival of breast cancer in Korean women. PMID- 15894658 TI - Association of breast cancer risk with a common functional polymorphism (Asp327Asn) in the sex hormone-binding globulin gene. AB - Sex hormones play a central role in the development of breast cancer. Sex hormone binding globulin (SHBG) modulates the bioavailability of circulating sex hormones and regulates their signaling system in the breast tissue. We evaluated the association of a common functional polymorphism (Asp327Asn) in the SHBG gene with breast cancer risk in a population-based case-control study (1,106 cases and 1,180 controls) conducted in Shanghai, China. The variant Asn allele was associated with a reduced breast cancer risk in postmenopausal women [odds ratio (OR), 0.73; 95% confidence interval (95% CI), 0.53-0.99], but not in premenopausal women (OR, 1.03; 95% CI, 0.82-1.27). The protective association was much stronger in postmenopausal women with a low body mass index (BMI; OR, 0.46; 95% CI, 0.29-0.75) or waist-to-hip ratio (OR, 0.51; 95% CI, 0.32-0.83) than those with a high BMI or waist-to-hip ratio (P for interaction < 0.05). Furthermore, the association was stronger for estrogen receptor-positive (OR, 0.64; 95% CI, 0.42-0.98) than for estrogen receptor-negative breast cancer (OR, 0.85; 95% CI, 0.50-1.45). Among postmenopausal controls, blood SHBG levels were 10% higher in carriers of the variant Asn allele than noncarriers (P = 0.06). Postmenopausal control women with the Asn allele and low BMI or waist-to-hip ratio had 20% higher SHBG levels (P < 0.05). This study suggests that the Asn allele in the SHBG gene may be related to a reduced risk of breast cancer among postmenopausal women by increasing their blood SHBG levels. PMID- 15894659 TI - Investigation of genetic variants of genes of the hemochromatosis pathway and their role in breast cancer. AB - Iron overload has been noticed as a feature of human breast cancer. Cellular iron uptake is regulated by the hemochromatosis and transferrin receptor system, mutations of which cause the iron storage disease hereditary hemochromatosis. To understand the role of hemochromatosis and transferrin receptor system mutations in breast cancer, we analyzed 19 sequence variations at HFE, TFR1, TFR2, and FPN1 and compared genotype frequencies between cases and controls in a German population. There were 688 breast cancer patients and 724 population-based and age-matched controls. For genotyping, we applied the Hemochromatosis Strip Assay and TaqMan allelic discrimination analyses. In addition to genotype frequencies, we established frequencies of compound genotypes. The frequencies of HFE at His63Asp, Ser65Cys, and Cys282Tyr, and of TFR1 at Ser142Gly minor alleles in this German population were 15.9%, 1.8%, 5.6%, and 46.0%, respectively. No rare variants at 15 more loci at HFE, TFR2, and FPN1 were observed in breast cancer patients. There were no significant differences of allele and genotype frequencies between cases and controls. Triple and quadruple compound genotypes at HFE_His63_Cys282-TFR1_Ser142Gly and HFE_His63_Ser65_Cys282-TFR1_Ser142Gly showed a nonsignificant increase in cases. Although limited by low numbers, an increased prevalence of the HFE Tyr282 minor allele was observed in breast cancer cases with a high number of affected lymph nodes (P = 0.032). Our data suggest that variants of the hemochromatosis-transferrin receptor system have no direct effect on the incidence of breast cancer in Germany. Possible effects on tumor progression and prognosis remain elusive. PMID- 15894660 TI - A basal epithelial phenotype is more frequent in interval breast cancers compared with screen detected tumors. AB - Interval breast cancer reduce the effectiveness of mammography screening programs. We studied 95 interval cancers, diagnosed during 1996 to 2001 as part of the population-based Norwegian Breast Cancer Screening Program. These cases were matched on size (+/-2.0 mm) to 95 screen-detected breast cancers, and the tumors were compared by immunohistochemical methods using tissue microarrays. Patients with interval cancers were more likely to be younger [odds ratio (OR), 4.7; P = 0.0001], to have dense breasts (OR, 3.4; P = 0.004), and to have estrogen receptor-negative tumors (OR, 2.6, P = 0.01), and p53 expression was more frequent (OR, 4.0; P = 0.001). Notably, interval cancers were more likely to have a basal epithelial phenotype, in that expression of cytokeratin 5/6 (OR, 2.3; P = 0.04) and P-cadherin (OR, 2.5; P = 0.04) was more frequent in interval cases than in size-matched, screen-detected tumors. In a logistic regression model, p53 expression, age, and breast density were independent predictors of interval cancers. Our data suggest that breast cancers with a basal epithelial phenotype are more likely than nonbasal breast cancers to present between regular mammograms. PMID- 15894661 TI - A correlation study of organochlorine levels in serum, breast adipose tissue, and gluteal adipose tissue among breast cancer cases in India. AB - We used data from a breast cancer pilot study carried out in Kerala, India in 1997, for which organochlorine levels were measured in three biological media, blood serum, breast adipose tissue, and gluteal adipose tissue, of 37 fasting breast cancer cases (pretreatment). Our objective was to investigate the relationships between organochlorine concentrations in different biological media. Gas-liquid chromatography determined serum, breast adipose, and gluteal adipose tissue levels of dichlorodiphenyltricholorethane, dichlorodiphenyldichloroethane, beta-benzene hexachloride, and polychlorinated biphenyl (PCB) congeners, PCB-153 and PCB-180. Correlation plots were made and Spearman correlation coefficients (r) calculated for breast adipose tissue versus serum, gluteal adipose tissue versus serum, and breast adipose versus gluteal adipose tissue. We also examined paired ratios of all summary statistics. There were strong correlations among serum, breast adipose tissue, and gluteal adipose tissue concentrations for most organochlorines analyzed, one exception being gluteal versus serum for PCB-153. The correlations for all other comparisons ranged from r = 0.65 to 0.94. Serum (ng/g) versus adipose ratios approached 1:1 for most of the organochlorine pesticide comparisons and did not vary by summary statistic. To our knowledge, this is the first study to use three different media from fasting subjects and to comprehensively investigate the relationship between organochlorines measured across the three media for both organochlorine pesticides and PCBs. These data indicate that blood serum reflects the present body burden of a range of organochlorines to the same extent as adipose tissue, and they support the view that serum may be collected in lieu of adipose tissue to obtain similar information. However, such measurements are a combination of both recent exposures and past exposures, which have metabolized slowly and may still persist. Therefore, investigators should use caution when assigning a level as lifetime body burden. PMID- 15894662 TI - Conditions associated with antibodies against the tumor-associated antigen MUC1 and their relationship to risk for ovarian cancer. AB - Many cancers, including ovarian, overexpress epithelial mucin (MUC1) and promote anti-MUC1 antibodies that may correlate with more favorable prognosis. By extension, risk for ovarian cancer might be reduced by preexisting MUC1-specific immunity. We measured anti-MUC1 antibodies in 705 control women, identified events predicting antibodies, and estimated ovarian cancer risk by comparing profiles of events generating antibodies in controls with those in 668 ovarian cancer cases. Factors predicting antibodies included oral contraceptive use, breast mastitis, bone fracture or osteoporosis, pelvic surgeries, nonuse of talc in genital hygiene, and to a lesser extent intrauterine device use and current smoking. There was a significant increase in the likelihood of having anti-MUC1 antibodies from 24.2% in women with 0 or 1 condition, to 51.4% in those with five or more conditions. By the same index of events, the risk for ovarian cancer was inversely associated with number of conditions predisposing to anti-MUC1 antibodies. Compared with having experienced 0 or 1 event, the adjusted risk for ovarian cancer decreased progressively with relative risks (and 95% confidence limits) of 0.69 (0.52-0.92), 0.64 (0.47-0.88), 0.49 (0.34-0.72), and 0.31 (0.16 0.61), respectively for women with two, three, four, and five or more events related to the presence of antibodies (P(trend) < 0.0001.) We conclude that several traditional and new risk factors for ovarian cancer may be explained by their ability to induce MUC1 immunity through exposure of MUC1 to immune recognition in the context of inflammatory or hormonal processes in various MUC1 positive tissues. PMID- 15894663 TI - Endometrial cancer incidence trends in Europe: underlying determinants and prospects for prevention. AB - More than one in 20 female cancers in Europe are of the endometrium. Surveillance of incidence rates is imperative given the rapidly changing profile in the prevalence and distribution of the underlying determinants. This study presents an analysis of observed and age-period-cohort-modeled trends in 13 European countries. There were increasing trends among postmenopausal women in many Northern and Western countries. Denmark and possibly France and Switzerland were exceptions, with decreasing trends in postmenopausal women. In premenopausal and perimenopausal women, declines were observed in Northern and Western Europe, most evidently in Denmark, Sweden, and the United Kingdom, affecting consecutive generations born after 1925. These contrast with the increasing trends regardless of menopausal age in some Southern and Eastern European countries, particularly Slovakia and Slovenia. These observations provide evidence of changes in several established risk factors over time and have implications for possible primary prevention strategies. In postmenopausal women, changes in reproductive behavior and prevalence of overweight and obesity may partially account for the observed increases, as well as hormone replacement therapy use in certain countries. Combined oral contraceptive use may be responsible for the declines observed among women aged <55 years. Whereas there are some prospects for chemoprevention in premenopausal women as oral contraceptive use becomes more widespread in Europe, increases in obesity and decreases in fertility imply that endometrial cancer in postmenopausal women will become a more substantial public health problem in the future. PMID- 15894664 TI - Physician recommendation for papanicolaou testing among U.S. women, 2000. AB - OBJECTIVE: Many women in the U.S. undergo routine cervical cancer screening, but some women have rarely or never had a Papanicolaou (Pap) test. Studies of other cancer screening tests (for example, mammograms) have shown that physician recommendation to get a screening test is one of the strongest predictors of cancer screening. METHODS: In this study, we examined whether women in the U.S. had received a physician recommendation to get a Pap test using data from the 2000 National Health Interview Survey. Reported reasons for not receiving a Pap test were also explored. RESULTS: Among women aged > or =18 years who had no history of hysterectomy, 83.3% [95% confidence interval (CI), 82.4-84.1%] of the 13,636 women in this sample had had a Pap test in the last 3 years. Among 2,310 women who had not had a recent Pap test, reported reasons for not receiving a Pap test included: "No reason/never thought about it" (48.0%; 95% CI, 45.5-50.7), "Doctor didn't order it" (10.3%; 95% CI, 8.7-12.0), "Didn't need it/didn't know I needed this type of test" (8.1%; 95% CI, 6.7-9.6), "Haven't had any problems" (9.0%; 95% CI, 7.6-10.5), "Put it off" (7.4%; 95% CI, 6.2-8.7), "Too expensive/no insurance" (8.7%; 95% CI, 7.3-10.2), "Too painful, unpleasant, embarrassing" (3.5%; 95% CI, 2.5-4.6), and "Don't have doctor" (1.7%; 95% CI, 1.2-2.4). Among women who had had a doctor visit in the last year but who had not had a recent Pap test, about 86.7% (95% CI, 84.5-88.6) reported that their doctor had not recommended a Pap test in the last year. African-American women were as likely as White women to have received a doctor recommendation to get a Pap test. Hispanic women were as likely as non-Hispanic women to have received a doctor recommendation to get a Pap test. In multivariate analysis, factors positively associated with doctor recommendation to get a Pap test included being aged 30 to 64 years, having been born in the U.S., and having seen a specialist or general doctor in the past year. CONCLUSION: These findings suggest that lack of a physician recommendation contributes to underuse of Pap screening by many eligible women. Given research that shows the effectiveness of physician recommendations in improving use, increased physician recommendations could contribute significantly to increased Pap screening use in the U.S. PMID- 15894665 TI - Modifiable risk factors associated with clearance of type-specific cervical human papillomavirus infections in a cohort of university students. AB - BACKGROUND: Previous findings regarding risk factors for human papillomavirus (HPV) persistence, other than viral determinants, identified from prospective cohort studies have been inconsistent in part because study designs have differed with respect to differing HPV detection methods and varying lengths of follow-up time. Therefore, the objectives of this study were to continue the search for epidemiologic risk factors of persistent cervical HPV infections and determine what behaviors differed between those women with transient HPV infections and those women who cannot clear their type-specific HPV infections. METHODS: Female university students (n = 621) in Montreal were followed for 24 months at 6-month intervals. At each visit, a cervical cell specimen was collected. HPV DNA was detected using the MY09/MY11 PCR protocol and 27 HPV genotypes were identified by the line blot assay (Roche Molecular Systems, Inc., Alameda, CA). Proportional hazards regression was used to estimate the crude and adjusted hazard ratios of clearing a type-specific high-risk (n = 222) or low-risk (n = 105) HPV infection over time according to specific baseline and time-dependent covariates. RESULTS: Daily consumption of vegetables seemed to increase the rate of HPV clearance independent of type. The use of tampons was associated with a reduced rate of high-risk HPV clearance, whereas regular condom use was associated with an increased rate of low-risk HPV clearance only. CONCLUSION: Some proactive measures can be taken to increase the rate of HPV clearance, and there may be some differences between the sets of predictors of low-risk and high-risk HPV clearance. PMID- 15894666 TI - Human papillomavirus genotype distribution in low-grade cervical lesions: comparison by geographic region and with cervical cancer. AB - Low-grade squamous intraepithelial lesions (LSIL) associated with certain human papillomavirus (HPV) genotypes may preferentially progress to cervical cancer. HPV genotyping may thus have the potential to improve the effectiveness of screening programs and to reduce overtreatment. LSIL cases (n = 8,308) from 55 published studies were included in a meta-analysis. HPV genotype distribution was assessed by geographic region and in comparison with published data on cervical squamous cell carcinoma (SCC). HPV detection in LSIL was 80% in North America but less than 70% in other regions, most likely reflecting regional differences in LSIL diagnosis. Among 5,910 HPV-positive LSILs, HPV16 was the most common genotype (26.3%) followed by HPV31 (11.5%), HPV51 (10.6%), and HPV53 (10.2%). HPV positive LSILs from Africa were 2-fold less likely to be infected with HPV16 than those in Europe, and HPV-positive LSILs from North America were more likely to be infected with HPV18 than those from Europe or South/Central America. Interpretation for rarer genotypes was hampered by variation in HPV testing methodology. SCC/LSIL prevalence ratios indicated that HPV16 was 2-fold and HPV18 was 1.5-fold more common in SCC than in HPV-positive LSIL, thus appearing more likely to progress than other high-risk genotypes (SCC/LSIL prevalence ratios between 0.05 and 0.85). HPV53 and HPV66 showed SCC/LSIL ratios of 0.02 and 0.01, respectively. HPV genotype distribution in LSIL differs from that in cervical cancer, highlighting the importance of HPV genotype in the risk of progression from LSIL to malignancy. Some regional differences in the relative importance of HPV genotypes in LSIL were noted. PMID- 15894667 TI - Smoking is a risk factor for cervical intraepithelial neoplasia grade 3 among oncogenic human papillomavirus DNA-positive women with equivocal or mildly abnormal cytology. AB - BACKGROUND: Smoking is a potential risk factor for cervical cancer and its immediate precursor, cervical intraepithelial neoplasia grade 3 (CIN3), but few studies have adequately taken into account the possible confounding effect of oncogenic human papillomavirus (HPV) infection. METHODS: Women (n = 5,060) with minimally abnormal Papanicolaou smears were enrolled in the ASCUS and LSIL Triage Study, a clinical trial to evaluate management strategies, and were seen every 6 months for the 2-year duration of the study. Cervical specimens were tested for HPV DNA using both Hybrid Capture 2 and PGMY09/11 L1 consensus primer PCR with reverse line blot hybridization for genotyping. Multivariate logistics regression models were used to assess associations [odds ratio (OR) with 95% confidence intervals (95% CI)] between smoking behaviors and rigorously reviewed cases of cervical intraepithelial neoplasia grade 3 or cancer (> or =CIN3) identified throughout the study (n = 506) in women with oncogenic HPV (n = 3,133). RESULTS: Current smoking was only weakly associated with increased HPV infection. Among infected women, current smokers (OR, 1.7; 95% CI, 1.4-2.1) and past smokers (OR, 1.7; 95% CI, 1.2-2.4) were more likely to be diagnosed with > or =CIN3 than nonsmokers. Greater smoking intensity (P(Trend) < 0.0005) and duration (P(Trend) < 0.0005) increased the strength of the association, with smoking > or =2 packs/d (OR, 3.3; 95% CI, 1.5-7.5) and smoking for > or =11 years (OR, 2.1; 95% CI, 1.5 2.9) most strongly associated with > or =CIN3 as compared to non-smokers. The effects of intensity and duration seemed additive. CONCLUSIONS: Women with oncogenic HPV and minimally abnormal Papanicolaou smears who smoke were up to three times more likely to be diagnosed with > or =CIN3 than nonsmokers. Smoking cessation trials targeting this population might be warranted. PMID- 15894668 TI - Multistage carcinogenesis and lung cancer mortality in three cohorts. AB - Experimental evidence indicates that tobacco smoke acts both as an initiator and a promoter in lung carcinogenesis. We used the two-stage clonal expansion model incorporating the ideas of initiation, promotion, and malignant conversion to analyze lung cancer mortality in three large cohorts, the British Doctors' cohort and the two American Cancer Society cohorts, to determine how smoking habits influence age-specific lung cancer rates via these mechanisms. Likelihood ratio tests indicate that smoking-related promotion is the dominant model mechanism associated with lung cancer mortality in all cohorts. Smoking-related initiation is less important than promotion but interacts synergistically with it. Although no information on ex-smokers is available in these data, the model with estimated variables can be used to project risks among ex-smokers. These projected risks are in good agreement with the risk among ex-smokers derived from other studies. We present 10-year projected risks for current and former smokers adjusted for competing causes of mortality. The importance of smoking duration on lung cancer risk in these cohorts is a direct consequence of promotion. Intervention and treatment strategies should focus on promotion as the primary etiologic mechanism in lung carcinogenesis. PMID- 15894669 TI - Mortality from lung cancer and tobacco smoking in Ohio (U.S.): will increasing smoking prevalence reverse current decreases in mortality? AB - BACKGROUND: Despite significant changes in smoking patterns within the past few decades, lung cancer remains a major cause of cancer deaths in many developed countries in people of each sex, and one of the most important public health issues. The study aims to analyze the possible impact of changes in tobacco smoking practices in the state of Ohio (U.S.) on current and future trends and patterns of lung cancer mortality. MATERIALS AND METHODS: Mortality rates from lung cancer were calculated for the period 1970 to 2001 on the basis of data from the National Center for Health Statistics. The Joinpoint regression approach was used to evaluate changes in time trends by sex, age, and race. Data on smoking prevalence in Ohio were retrieved from the Centers for Disease Control and Prevention website. RESULTS: Lung cancer mortality rates in Ohio have declined among men of all ages as well as in specific age groups in the 1990s, and the rate of increase among middle-aged and elderly women has dropped over time. The mortality rate among young women (ages 20-44) began to increase during the early 1990s. The prevalence of smoking in Ohio has increased since the early 1990s, especially among young persons. CONCLUSIONS: Recent trends in tobacco smoking in Ohio indicate that the declining trends in lung cancer mortality might be reversed in the future. An early indicator of possible change is the recent increase in mortality among young women. Implementation of the Ohio Comprehensive Tobacco Use Prevention Strategic Plan might help to disseminate proven prevention strategies among the inhabitants of Ohio and might thus prevent future increases in lung cancer mortality rates in the state. PMID- 15894670 TI - Polymorphisms of methionine synthase and methionine synthase reductase and risk of squamous cell carcinoma of the head and neck: a case-control analysis. AB - Although tobacco and alcohol use are the major risk factors, folate deficiency has been implicated in the risk of squamous cell carcinoma of the head and neck (SCCHN). We hypothesized that polymorphisms of methionine synthase (MTR) and methionine synthase reductase (MTRR) in the folate metabolic pathway are associated with SCCHN risk. In a hospital-based case-control study of 721 SCCHN cases and 1,234 controls of non-Hispanic Whites, frequency matched by age, sex, and smoking status, we genotyped the MTR A2756G and MTRR G66A polymorphisms. We found that the MTR variant AG and AG/GG genotypes were associated with a significantly increased SCCHN risk [adjusted odd ratio (OR), 1.31; 95% confidence interval (95% CI), 1.07-1.60 for AG and OR, 1.28; 95% CI, 1.05-1.56 for AG/GG] compared with the AA genotype. In contrast, the MTRR variant AA genotype was associated with a significantly decreased SCCHN risk (OR, 0.68; 95% CI, 0.52 0.90) compared with the 66GG genotype. When the two polymorphisms were evaluated together by the number of risk alleles, the SCCHN risk was significantly increased in a dose-dependent manner (P(trend) = 0.002). The risk of SCCHN was 1.47 (95% CI, 1.08-1.99) for one risk allele, 1.67 (95% CI, 1.23-2.27) for two risk alleles, and 1.74 (95% CI, 1.18-2.54) for three or four risk alleles compared with the wild-type (0 risk allele) genotype. In conclusion, our data provide evidence that support the association between the MTR A2756G and MTRR G66A polymorphisms and SCCHN risk and that these two polymorphisms may have a joint effect on risk of SCCHN. PMID- 15894671 TI - No association between gastroesophageal reflux and cancers of the larynx and pharynx. AB - PURPOSE: To examine the proposed relation between gastroesophageal reflux disease and cancers of the larynx and pharynx. EXPERIMENTAL DESIGN: A cohort of 66,965 patients with discharge diagnoses of heartburn, hiatal hernia, or esophagitis was identified in the Swedish Inpatient Register. We observed a total of 376,622 person-years in the cohort. Identification of laryngeal and pharyngeal cancers was achieved through the Swedish Cancer Register. Standardized incidence ratios, the ratio of the observed to the expected number of cancers, derived from the general Swedish population and adjusted for sex, age, and calendar year, was used to estimate the relative risk. RESULTS: During follow-up, 30 cases of laryngeal and 31 cases of pharyngeal cancer were detected in the cohort. Slightly increased risks of laryngeal [relative risk, 1.6; 95% confidence interval (CI), 1.1-2.2] and pharyngeal carcinomas (relative risk, 1.9; 95% CI, 1.3-2.8) were observed in the total reflux cohort. After stratification to exclude cohort members with a diagnosis of alcoholism, no significant increase in the risk of laryngeal (relative risk, 1.3; 95% CI, 0.8-2.0) or pharyngeal carcinomas (relative risk, 1.0; 95% CI, 0.5-1.6) was found compared with the general population. Furthermore, there were no dose-response patterns of the risk for laryngeal and pharyngeal cancers with years of cohort follow-up, indicators of reflux severity, or specificity of reflux diagnosis. CONCLUSION: This study provides no evidence in support of the proposed association between gastroesophageal reflux disease and cancers of the larynx or pharynx. PMID- 15894672 TI - The MTHFR C677T polymorphism and colorectal cancer: the multiethnic cohort study. AB - Methylenetetrahydrofolate reductase (MTHFR) is a key regulatory enzyme in the metabolism of folate, a nutrient that has been inversely related to colorectal cancer risk. The common C677T variant in the MTHFR gene results in a reduced activity of this enzyme, thereby increasing the availability of folate for the production of thymidylate and purine for DNA synthesis and repair. We investigated the association of the 677TT genotype with colorectal cancer in a case-control study of 822 cases and 2,021 controls nested within the Multiethnic Cohort Study. The Multiethnic Cohort Study is a large prospective study of men and women of Japanese, White, African American, Latino, and Native Hawaiian origin, residing in Hawaii and Los Angeles. After adjusting for covariates, we found an inverse association between colorectal cancer risk and the TT genotype, with odds ratios (OR; and 95% confidence intervals) for the CC, CT, and TT genotypes of 1.00, 1.01 (0.84-1.21), and 0.77 (0.58-1.03), respectively. This association was similar in both sexes, stronger at high levels of folate intake, and limited to light and nondrinkers (P for interaction with ethanol = 0.02). An analysis by subsite (rectum versus colon) and stage (regional/distant versus in situ/localized) showed that the inverse association with the TT genotype was limited to colon tumors, especially those diagnosed at an advanced stage. The OR for the TT versus CC genotype for early- and late-stage colon cancer was 0.88 (0.58-1.33) and 0.52 (0.32-0.85), respectively (P for difference in OR = 0.04). The frequency of the T allele was relatively low in African Americans (0.13) and Native Hawaiians (0.22), consistent with their greater likelihood of presenting at a late stage when diagnosed with colorectal cancer. This study corroborates previous findings of an inverse association of the MTHFR 677TT genotype with colorectal cancer, especially at high levels of folate and low levels of ethanol intake. It also suggests that this effect may be specific to advanced colon cancer. PMID- 15894673 TI - Insulin-like growth factor polymorphisms and colorectal cancer risk. AB - Several modifiable lifestyle factors, such as physical activity, obesity, and postmenopausal hormone use, have been associated with colorectal cancer risk. It has been hypothesized that some or all of these factors may mediate their effects through alterations in insulin-like growth factor-1 (IGF-1) and its binding proteins (IGFBP). To evaluate the role of IGFs in colorectal cancer, we examined the relationship of two common genetic polymorphisms in IGF-1 (a cytosine adenosine dinucleotide repeat) and IGFBP-3 (a G --> C single nucleotide polymorphism) with colorectal cancer risk, as well as their potential modification by physical activity, body mass index (BMI), and postmenopausal hormone use. Subjects included 782 male and female colorectal cancer cases diagnosed between 1998 and 2002 and reported to the statewide registry in the metropolitan Seattle area, and 503 age- and sex-matched cancer-free population controls. Colorectal cancer was modestly associated with having an IGF-1 genotype other than homozygous for 19 repeats (odds ratio, 1.3; 95% confidence interval, 1.0-1.6) and having the GG IGFBP-3 genotype (odds ratio, 1.3; 95% confidence interval, 1.0-1.8). There was evidence that IGF-1 genotype modified the relationship between BMI and colorectal cancer among women, such that high BMI increased risk of colorectal cancer only among those with the 19/19 genotype (P(interaction) = 0.02). IGFBP-3 genotype was also a significant effect modifier of the relationship between risk factors and colorectal cancer: The positive association between BMI and colorectal cancer was observed only among men (P(interaction) < 0.01) and women (P(interaction) = 0.06) with the GG genotype; the inverse association between postmenopausal hormone use and colorectal cancer was observed only among women with the GG genotype (P = 0.01) and the inverse association between physical activity and colorectal cancer was observed only among men who carried the C allele (P < 0.01). The current study provides some support for a role of IGFs in colorectal cancer etiology, particularly in mediating the relationship of common risk factors (physical activity, BMI, and postmenopausal hormone use). PMID- 15894674 TI - Oral contraceptive use, reproductive factors, and colorectal cancer risk: findings from Wisconsin. AB - We investigated the association of oral contraceptive (OC) use and reproductive factors with colorectal cancer risk in a large population-based case-control study. Cases were women ages 20 to 74 years, living in Wisconsin, with a new diagnosis of colon (n = 1,122) or rectal (n = 366) cancer. Control participants were randomly selected from population lists of similarly aged female Wisconsin residents (n = 4,297). Risk factor information was collected through structured telephone interviews. Compared with never users, OC users had an odds ratio (OR) of 0.89 [95% confidence interval (95% CI), 0.75-1.06] for colorectal cancer. OC use associations did not differ significantly between colon and rectal cancer sites; however, when compared with never users, recent OC users (<14 years) seemed at reduced risk of rectal cancer (OR, 0.53; 95% CI, 0.28-1.00). Women with age at first birth older than the median (23 years) had 0.83 times the risk of colon cancer compared with women with age at first birth below the median (95% CI, 0.70-0.98). We observed an inverse trend between increasing parity and rectal cancer risk (P = 0.05). Compared with nulliparous women, women with five or more births had 0.66 times the risk of rectal cancer (95% CI, 0.43-1.02). Compared with postmenopausal women, premenopausal women were at reduced risk (OR, 0.67; 95% CI, 0.47-0.97) of colorectal cancer. No significant associations were observed between colorectal cancer risk and age at menarche or age at menopause. These findings suggest differential roles of reproductive factors in colon and rectal cancer etiology. PMID- 15894675 TI - Gene-specific methylation and subsequent risk of colorectal adenomas among participants of the polyp prevention trial. AB - Hypermethylation of tumor suppressor and other regulatory genes is thought to play an important role in colorectal neoplasia and tumorigenesis. This study examined the association between gene methylation status in baseline adenomas and subsequent adenoma recurrence in a randomized dietary intervention study, the Polyp Prevention Trial. The methylation status of four genes [CDKN2A (p16), PTGS2 (COX2), ESR1 (ER-alpha), and PGR(PR)] was determined by MethyLight in 284 baseline adenomas from 196 trial participants. The association of gene methylation with recurrence was determined using logistic regression models. Gene methylation was evaluated as percent of methylated reference, a measure of methylation of each gene relative to control DNA. ESR1methylation status was inversely associated with adenoma recurrence, odds ratio = 0.36 (95% confidence interval, 0.15-0.88; P = 0.02) for the highest compared with the lowest quartile of the ESR1methylation. Further, ESR1 methylation status was inversely associated with the recurrence of multiple adenomas, advanced adenomas, and the recurrence of adenomas in the proximal but not distal bowel. No association between CDKN2A, PTGS2, or PGR methylation and adenoma recurrence was observed. These data suggest that ESR1 methylation may play a role in subsequent adenoma recurrence. PMID- 15894676 TI - Interactions of peroxisome proliferator-activated receptor {gamma} and diet in etiology of colorectal cancer. AB - The peroxisome proliferator-activated receptor gamma (PPARgamma) is one of a group of ligand-activated nuclear receptors responsible for regulation of glucose, lipid homeostasis, cell differentiation, and apoptosis. The 12 proline to-alanine (Pro12Ala) substitution polymorphism in PPARgamma produces proteins with lower activity. Variation in PPARgamma expression in the bowel and the role of dietary fatty acids as ligands for PPARgamma led investigation of whether the associations of diet with colon and rectal cancer risk were modified by PPARgamma genotype. Data (diet, lifestyle, and DNA) came from case-control studies of colon (1,577 cases and 1,971 controls) and rectal cancer (794 cases and 1,001 controls) conducted in Northern California, Utah, and the Twin City, Minnesota Metropolitan area (colon cancer study only). Unconditional logistic regression models were adjusted for age at selection, body mass index, physical activity, energy intake, dietary fiber, and calcium. We found no significant interactions between macronutrient (fat, protein, and carbohydrate) and colorectal cancer. High lutein intake [odds ratio (OR), 0.63; 95% confidence interval (95% CI), 0.44-0.89], low refined grain intake (OR, 0.70; 95% CI, 0.53-0.94), or a high prudent diet score (OR, 0.66; 95% CI, 0.49-0.89) and PA/AA PPARgamma genotype were associated with reduced colon cancer risk. Risk of rectal cancer was increased among those with the PA/AA PPARgamma genotype and a high mutagen index (OR, 1.63; 95% CI, 1.12, 2.36). Its unclear whether the alterations in risk in those with the less active phenotype for PPARgamma is related to activation of PPARgamma by nutrients or dietary patterns acting as ligands or direct influences of these nutrients on colon and rectal cancer processes that are important with lower PPARgamma activity. PMID- 15894677 TI - Tissue-specific attenuation of endogenous DNA I-compounds in rats by carcinogen azoxymethane: possible role of dietary fish oil in colon cancer prevention. AB - I-compounds are bulky covalent DNA modifications that are derived from metabolic intermediates of nutrients. Some I-compounds may play protective roles against cancer, aging, and degenerative diseases. Many carcinogens and tumor promoters significantly reduce I-compound levels gradually during carcinogenesis. Colon cancer is the second leading cause of cancer death in the United States, whereas cancer of the small intestine is relatively rare. Here we have studied levels of I-compounds in DNA of colon and duodenum of male Sprague-Dawley rats treated with azoxymethane. The effects of dietary lipids (fish oil or corn oil) on colon and duodenal DNA I-compounds were also investigated. Rats fed a diet containing fish oil or corn oil were treated with 15 mg/kg azoxymethane. Animals were terminated 0, 6, 9, 12, or 24 hours after injection. I-compound levels were analyzed by the nuclease P1-enhanced (32)P-postlabeling assay. Rats treated with azoxymethane displayed lower levels of I-compounds in colon DNA compared with control groups (0 hour). However, I-compound levels in duodenal DNA were not diminished after azoxymethane treatment. Animals fed a fish oil diet showed higher levels of I compounds in colonic DNA compared with corn oil groups (mean adduct levels for fish and corn oil groups were 13.35 and 10.69 in 10(9) nucleotides, respectively, P = 0.034). Taken together, these results support claims that fish oil, which contains a high level of omega-3 polyunsaturated fatty acids, may have potent chemopreventive effects on carcinogen-induced colon cancer. The fact that duodenal I-compounds were not diminished by azoxymethane treatment may have been due to the existence of tissue-specific factors protecting against carcinogenesis. In conclusion, our observations show that endogenous DNA adducts may serve not only as sensitive biomarkers in carcinogenesis and cancer prevention studies, but are also helpful to further our understanding of the chemopreventive properties of omega-3 fatty acids and mechanisms of carcinogenesis. PMID- 15894678 TI - Childhood and adult cancer in twins: evidence from the Utah genealogy. AB - OBJECTIVE: Evidence suggests that the in utero environment may contribute to subsequent development of cancers in childhood and adulthood. Raised levels of estrogen during pregnancy may be the primary in utero etiologic factor. Mothers of twins have higher estrogen levels during pregnancy than mothers of singletons, therefore, assessment of cancer risk in twins may be informative. METHOD: We conducted a retrospective cohort study of cancer among twin and singleton newborns selected from the Utah Population Database, matched on birth year and sex. Cancer diagnoses were determined by linkage with the Utah state cancer register. Relative rates of all cancers in childhood and in adulthood in twins compared with singletons, and for specific cancers including testicular, breast and melanoma, were calculated using Poisson regression. RESULTS: Twin (35,271) and singleton (74,199) births were identified, among whom there were 336 and 691 cancer diagnoses, respectively. The relative risk (RR) of childhood cancer in twins compared with singletons was 0.82 [95% confidence interval (CI) 0.55-1.24] and of adult cancer was 1.06 (0.92-1.22). We found nonsignificant increases in risk among adult twins for cancers of the breast, prostate, testis, lymphatic system, thyroid, and large bowel. The largest departures from unity were for testicular cancer (RR 1.47; 95% CI, 0.73-2.95) and melanoma (RR 0.67; 95% CI, 0.42-1.06). CONCLUSIONS: These results are consistent with the body of evidence suggesting that twins have a reduced risk of cancer in childhood. Although there is no overall differential in adult cancer risk, these data support the hypothesis that the in utero environment may play an important role in specific cancers. PMID- 15894679 TI - Risk factors for melanoma by body site. AB - It has been hypothesized that cutaneous melanoma at different anatomic sites develops through divergent pathways. We examined this hypothesis prospectively. We followed 152,949 women and 25,204 men free of cancer at baseline for up to 14 years in three large prospective studies. We examined risk factors for melanoma by anatomic location (head or neck, trunk, upper extremity, and lower extremity). Polytomous logistic regression was used to test the difference among risk factors by location of melanoma. A total of 511 incident cases of invasive melanoma (49 head or neck, 188 trunk, 98 upper extremity, and 176 lower extremity) were included in the analysis. Compared with females, males had a higher risk of developing melanoma on the head or neck and trunk. History of severe and painful sunburn was most strongly related to melanoma of upper extremity; individuals with >10 burns had a 6.86-fold (95% confidence interval, 2.62-18.00) higher risk of melanoma of upper extremity compared with those with no burns (P for trend < 0.0001; P for difference by body site = 0.04). Number of moles was most strongly related to melanoma of the trunk; the multivariate relative risk for having >10 moles was 4.67 (95% confidence interval, 3.07-7.11) compared with having no moles (P for trend < 0.0001; P for difference by body site = 0.04). Age, family history of melanoma, and hair color did not statistically differ by anatomic site of the cancer. These data support divergent etiologic pathways of melanoma development by anatomic sites. PMID- 15894680 TI - No evidence for anticipation in lymphoproliferative tumors in population-based samples. AB - Genetic anticipation in familial non-Hodgkin's lymphoma, Hodgkin's lymphoma, and chronic lymphocytic leukemia (CLL) has been consistently reported in the literature. However, most of these findings were based on data from families ascertained for genetic studies. Fecundity bias, right censoring bias, and secular trends can lead to erroneous conclusions regarding the presence of anticipation. Our report investigates anticipation in four lymphoproliferative cancers, non-Hodgkin's lymphoma, Hodgkin's lymphoma, CLL, and multiple myeloma, drawn from Swedish and Danish population-based registries. We used marginal survival methods to test for a relative difference in age at diagnosis between parents and offspring and to account for other risk factors, staggered entries, censored data, and correlations among relatives. Changes in incidence rates of lymphoproliferative tumors were accommodated in the models by using time-varying covariates for different periods of diagnosis. Whereas no anticipation was observed for Hodgkin's lymphoma, CLL, and multiple myeloma, our initial model, which controlled for gender and country, suggested a significant difference (hazard ratio, 0.5; 95% confidence interval, 0.33-0.75) in age at diagnosis between the parents and offspring in the non-Hodgkin's lymphoma sample. However, once we accounted for the significant change in non-Hodgkin's lymphoma incidence over time, the statistical difference between parents and offspring disappeared (hazard ratio, 0.99; 95% confidence interval, 0.56-1.76). Our results emphasize the importance of considering secular trends when evaluating the possibility of anticipation in lymphoproliferative cancers. This is the first study to consider the changes of incidence over time as a source of bias when evaluating anticipation in lymphoproliferative cancers. PMID- 15894681 TI - Polymorphisms in ghrelin and neuropeptide Y genes are associated with non-Hodgkin lymphoma. AB - We previously reported a positive association among body mass index, single nucleotide polymorphisms (SNP) in the leptin and leptin receptor genes that are involved in body weight regulation, and non-Hodgkin lymphoma (NHL). Polymorphisms in the ghrelin (GHRL) and neuropeptide Y (NPY) genes were examined in the same population-based case-control study of NHL to further explore the role of genes involved in energy homeostasis and obesity in susceptibility to NHL. Ghrelin is an orexigenic hormone that induces NPY release and inhibits proinflammatory cytokines via its antagonistic relationship with leptin. NPY is a potent appetite stimulator controlled by ghrelin and leptin and also acts as a mediator of immune function. DNA from 458 cases and 812 controls was genotyped. Among genotyped GHRL SNPs, the variant allele for GHRL -4427G>A was inversely associated with all NHL [odds ratios (OR), 0.78; 95% confidence interval (95% CI), 0.59-1.0] and more specifically with diffuse large cell lymphoma (DLCL; homozygous variant: OR, 0.31; 95% CI, 0.13-0.74). Another SNP, GHRL 5179A>G, decreased the risk of DLCL (homozygous variant: OR, 0.35; 95% CI, 0.10-1.2). NPY -485T>C, 1258G>A, and 5671C>T were in total linkage disequilibrium (D' = 0.99) and the homozygous variants were associated with an increased risk of NHL in NPY SNPs -485T>C (OR, 1.7; 95% CI, 1.1-2.5), 1258G>A (OR, 1.7; 95% CI, 1.1-2.5), and 5671C>T (OR, 1.9; 95% CI, 1.3-2.8). When stratified by subtype, the variant allele for NPY 1128T>C was positively associated with follicular lymphoma (OR, 2.3; 95% CI, 1.1-4.9) as were homozygous variants for NPY SNPs -485T>C (OR, 2.4; 95% CI, 1.3-4.4), 1258G>A (OR, 2.0; 95% CI, 1.1-3.5), and 5671C>T (OR, 1.8; 95% CI, 1.1-3.0). These results add further support for the hypothesis that SNPs in energy-regulating genes affect risk of NHL. PMID- 15894682 TI - CYP3A43 Pro(340)Ala polymorphism and prostate cancer risk in African Americans and Caucasians. AB - The human cytochrome P450 3A subfamily of enzymes is involved in the metabolism of steroid hormones, carcinogens, and many drugs. A cytosine-to-guanine polymorphism in CYP3A43 results in a proline-to-alanine substitution at codon 340. Although the functional significance of this polymorphism is unknown, we postulate that the substitution of proline, an alpha-imino acid, with alanine, an amino acid, could be of biochemical significance. In a case-control study with 490 incident prostate cancer cases (124 African Americans and 358 Caucasians) and 494 controls (167 African Americans and 319 Caucasians), we examined the association between CYP3A43 Pro(340)Ala polymorphism and prostate cancer risk. When all subjects were considered, there was a 3-fold increase in risk of prostate cancer among individuals with the CYP3A43-Ala/Ala genotype (odds ratio, 3.0; 95% confidence interval, 1.2-7.2) compared with those with the CYP3A43 Pro/Pro genotype after adjusting for age, race, and smoking. The prevalence of the polymorphism was significantly higher in African Americans than Caucasians (45% versus 13%). In African Americans, there was a 2.6-fold increase in prostate cancer risk among individuals with the CYP3A43-Ala/Ala genotype (odds ratio, 2.6; 95% confidence interval, 1.0-7.0) compared with those with the CYP3A43-Pro/Pro genotype. Among Caucasians, the small number of homozygotes precluded computing risk estimates; there were only three individuals with the CYP3A43-Ala/Ala genotype. Our results suggest that the CYP3A43-Pro(340)Ala polymorphism contributes to prostate cancer risk. PMID- 15894683 TI - Sex steroid hormones and the androgen receptor gene CAG repeat and subsequent risk of prostate cancer in the prostate-specific antigen era. AB - OBJECTIVE: Sex steroid hormones are thought to contribute to the growth, differentiation, and progression of prostate cancer. We investigated plasma levels of sex steroid hormones and length of the androgen receptor gene CAG repeat in relation to incident prostate cancer diagnosed in the prostate-specific antigen (PSA) era. METHODS: Using a nested case-control design, we included 460 prostate cancer cases diagnosed after providing a blood specimen in 1993 but before February 1998 among men in the Health Professionals Follow-up Study. Controls were 460 age-matched men without prostate cancer who had a screening PSA test after the date of providing a blood specimen. We measured plasma concentrations of total testosterone, free testosterone, dihydrotestosterone, androstanediol glucuronide, estradiol, and sex hormone binding globulin (SHBG) and determined the length of the androgen receptor gene CAG repeat. Conditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) of prostate cancer. RESULTS: Mean concentrations of the sex steroids adjusted for SHBG, and mean CAG repeat length did not differ significantly between the prostate cancer cases and controls. No significant associations with total prostate cancer were detected for plasma total testosterone concentration (comparing highest versus lowest quartiles: OR, 0.78; 95% CI, 0.48-1.28; P(trend) = 0.73) or the other sex hormones after adjusting for SHBG. However, plasma total testosterone concentration was positively associated with low-grade disease (Gleason sum < 7: OR, 1.91; 95% CI, 0.89-4.07; P(trend) = 0.02) and inversely associated with high-grade disease (Gleason sum > or = 7: OR, 0.26; 95% CI, 0.10-0.66; P(trend) = 0.01). Similar patterns for grade were observed for free testosterone. Short CAG repeat length was not associated with total prostate cancer (< or = 19 versus > or = 24: OR, 0.84; 95% CI, 0.57-1.23; P(trend) = 0.22) or grade of disease. No clear associations with regionally invasive or metastatic (> or = T3b, N1, or M1) were found for any of the hormones or the CAG repeat, although the number of these cases was small. CONCLUSIONS: The overall lack of association of prostate cancer diagnosed in the PSA era with sex steroid hormones and the androgen receptor gene CAG repeat length is consistent with the hypothesis that these factors do not substantially contribute to the development of early prostate cancer in the PSA era. The influence of plasma total and free testosterone concentrations on prostate cancer grade merits further evaluation. PMID- 15894684 TI - Serum insulin-like growth factor-I levels and prostatic intraepithelial neoplasia: a clue to the relationship between IGF-I physiology and prostate cancer risk. AB - Serum insulin-like growth factor-I (IGF-I) levels at the higher end of the reference range have been associated with increased risk for the future development of prostate cancer. We determined whether high serum IGF-I levels are associated with precancerous lesions of the prostate. We conducted a case-control study to determine whether high serum IGF-I levels were associated with the presence of high-grade prostatic intraepithelial neoplasia (HGPIN) among patients who presented for prostate biopsy because of an abnormal serum prostate-specific antigen level or digital rectal exam. We measured serum IGF-I and insulin-like growth factor binding protein-3 (IGFBP-3) prior to prostate biopsy and compared them between 103 men with HGPIN (cases) and 205 men with normal prostate histology (controls). The mean IGF-I level in patients with HGPIN (130.2 ng/mL) was significantly higher compared with controls (118.8 ng/mL, P = 0.01). The mean IGFBP-3 level in patients with HGPIN (2,393.9 ng/mL) was also higher compared with controls (2,276.0 ng/mL, P = 0.06). After adjusting for age, prostate specific antigen, digital rectal examination, and ethnic background, the odds ratio for a HGPIN diagnosis among men in the highest relative to the lowest quartile of serum IGF-I level was 1.94 (95% confidence interval, 1.0-3.7; P = 0.04). The potential association between a high serum IGF-I level and the presence of HGPIN may represent an important clue to understanding the basis for the relationship between IGF-I physiology and prostate cancer risk. Larger studies will be required to confirm this relationship. PMID- 15894685 TI - MT1G hypermethylation is associated with higher tumor stage in prostate cancer. AB - PURPOSE: Zinc is involved in several physiologic processes, including cell growth and proliferation. Although in normal prostate tissue zinc levels are high, there is a marked decrease in prostate cancer. Metallothioneins control the bioavailability of zinc and one isoform, MT1G, was reported down-regulated in prostate cancer. Here, we investigated whether promoter methylation might cause MT1G silencing in prostate cancer. PATIENTS AND METHODS: The MT1G promoter was assessed by quantitative methylation-specific PCR on prospectively collected tissue samples from 121 patients with prostate cancer, 39 paired high-grade prostatic intraepithelial neoplasias (HGPIN), 29 patients with benign prostatic hyperplasia, 13 normal prostate tissue samples from cystoprostatectomy specimens, and prostate cancer cell lines. The methylation levels were calculated and were correlated with clinical and pathologic variables. Reverse transcription-PCR was done in cell lines to assess MT1G mRNA expression before and after demethylating treatment. RESULTS: MT1G promoter hypermethylation was found in 29 of 121 prostate cancer, 5 of 39 HGPIN, 3 of 29 benign prostatic hyperplasia, and 0 of 13 normal prostate tissue samples. No significant differences in methylation frequencies or levels were found (P = 0.057, for both). Methylation levels were found to correlate with tumor stage but not with Gleason grade. MT1G hypermethylation was more frequent in prostate cancer that spread beyond the prostate capsule. All prostate cancer cell lines tested showed MT1G promoter methylation, but no differences in expression were apparent after demethylation. CONCLUSIONS: Our findings suggest that MT1G promoter methylation is associated with tumor aggressiveness in prostate cancer and it might be a marker of locally advanced disease. PMID- 15894686 TI - Tumor antigen markers for the detection of solid cancers in inflammatory myopathies. AB - Dermatomyositis and polymyositis patients have an increased risk of developing cancers. We have assessed the diagnostic values of serum tumor markers for the detection of solid cancer in dermatomyositis/polymyositis patients. Serum carcinoembryonic antigen, CA15-3, CA19-9, and CA125 were assayed by immunoradiometric methods in 102 dermatomyositis/polymyositis patients. All the patients had complete physical examination, chest X-ray, echocardiogram, gastrointestinal tract endoscopic explorations, thoracoabdomino-pelvic computed tomography scan, and all women had gynecologic examination and mammogram. Exclusion criteria for study were childhood dermatomyositis, inclusion body myositis, myositis associated with a connective tissue disease, prior history of cancer, and the presence of benign conditions known to elevate serum tumor markers. After a median follow-up of 59 months, 10 (9.8%) patients had a solid cancer. Initial elevation of CA125 was associated with an increased risk of developing solid cancer [P = 0.0001 by Fisher's exact test; odds ratio (OR), 29.7; 95% confidence interval (95% CI), 8.2-106.6]. For CA19-9, there was a trend towards a significant association (P = 00.7; OR, 4.5; 95% CI, 1-18.7, respectively). Diagnostic values of elevated CA125 and CA19-9 at screening increased when the study analysis was restricted to patients who developed a cancer within 1 year (P < 0.0001 and P = 0.018, respectively) or to patients without interstitial lung disease (P = 0.00001; OR, 133; 95% CI, 6.5-2733 and P = 0.027; OR, 9; 95% CI, 1.5-53, respectively). Individual comparisons of the baseline and the second CA125 value showed that three of the eight patients with cancers versus 3 of the 76 patients without, displayed an increase of their CA125 level (P = 0.01 by Fisher's exact test). We conclude that CA125 and CA19-9 assessment could be useful markers of the risk of developing tumors for patients with dermatomyositis and polymyositis and should therefore be included in the search for cancer in dermatomyositis/polymyositis patients, especially for patients without interstitial lung disease. PMID- 15894687 TI - Urinary metabolites of a tobacco-specific lung carcinogen in nonsmoking hospitality workers. AB - Exposure of nonsmokers to environmental tobacco smoke results in increased risk for cancer and other diseases. In spite of this finding, some restaurants and bars continue to permit smoking. This study examined the uptake of nicotine and 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a potent lung carcinogen, in nonsmokers who work in restaurants and bars that permitted smoking. Urine samples were collected for 24 hours on working and nonworking days and were analysed for total NNAL [the sum of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronides (NNAL-Glucs)], metabolites of NNK. In addition, urine samples were analysed for total nicotine (nicotine plus nicotine glucuronide), and total cotinine (cotinine plus cotinine-N-glucuronide). The results showed significant increases in urinary levels of total NNAL, total nicotine, and total cotinine on working days compared with nonworking days. The results of this study show that smoke exposure in bars and restaurants may have important health effects on nonsmoking employees, elicited by the increase in carcinogen levels. PMID- 15894688 TI - Urinary sucrose and fructose as biomarkers for sugar consumption. AB - The use of 24-hour urinary sucrose and fructose as potential biomarkers for sugars consumption was investigated in two studies of 21 healthy participants living in a volunteer suite where dietary intake was known and all specimens collected. The dose-response was assessed in 12 males using a randomized crossover design of three diets containing constant levels of 63, 143, and 264 g of sugars for 10 days each. Both sugars and sucrose intake were significantly correlated with the sum of sucrose and fructose concentration in urine (0.888; P < 0.001). To assess effects with volunteers consuming their habitual varying diets, seven males and six females were fed their usual diet (assessed beforehand from four consecutive self-completed 7-day food diaries) for 30 days under controlled conditions in the volunteer suite. The mean (+/-SD) calculated total sugars intake was 202 +/- 69 g/d, 41% from sucrose. Mean (+/-SD) urinary sucrose and fructose were 36.6 +/- 16.6 and 61.8 +/- 61.3 mg/d, respectively. The sum of sucrose and fructose in urine was significantly correlated with sugars (0.841; P < 0.001) and sucrose intake (0.773; P = 0.002). In the regression, 200 g of sugars intake predicted approximately 100 mg of sucrose and fructose in urine. The correlation between individual means of randomized 16 days of sugars intake and 8 days of sugars excretion data (as used in validation studies) remained as high as that obtained with the means of 30-day measurements and the regression estimates were very similar. Twenty-four-hour urinary sucrose and fructose could be grouped into a new category of biomarkers, predictive biomarkers, that can be used in studies determining the structure of dietary measurement error in free living individuals and to relate sugars intake to disease risk. PMID- 15894689 TI - The association of XRCC1 haplotypes and chromosomal damage levels in peripheral blood lymphocyte among coke-oven workers. AB - Theoretically, a haplotype has a higher level of heterozygosity than individual single nucleotide polymorphism (SNP) and the association study based on the haplotype may have an increased power for detecting disease associations compared with SNP-based analysis. In this study, we investigated the effects of four haplotype-tagging SNPs (htSNP) and the inferred haplotype pairs of the X-ray cross-complementing group 1 (XRCC1) gene on chromosome damage detected by the cytokinesis-block micronucleus assay. The study included 141 coke-oven workers with exposure to a high level of polycyclic aromatic hydrocarbons and 66 nonexposed controls. The frequencies of total MN and MNed cells were borderline associated with the Arg(194)Trp polymorphism (P = 0.053 and P = 0.050, respectively) but not associated with the Arg(280)His, Arg(399)Gln and Gln(632)Gln polymorphisms among coke-oven workers. Five haplotypes, including CGGG, TGGG, CAGG, CGAG, and CGGA, were inferred based on the four htSNPs of XRCC1 gene. The haplotype CGGG was associated with the decreased frequencies of total MN and MNed cells, and the haplotypes TGGG and CGAG were associated with the increased frequencies of total MN and MNed cells with adjustment for covariates among coke-oven workers. This study showed that the haplotypes derived from htSNPs in the XRCC1 gene were more likely than single SNPs to correlate with the polycyclic aromatic hydrocarbon-induced chromosome damage among coke-oven workers. PMID- 15894690 TI - Increased rates of chromosome breakage in BRCA1 carriers are normalized by oral selenium supplementation. AB - Women who are born with constitutional heterozygous mutations of the BRCA1 gene face greatly increased risks of breast and ovarian cancer. The product of the BRCA1 gene is involved in the repair of double-stranded DNA breaks and it is believed that increased susceptibility to DNA breakage contributes to the cancer phenotype. It is hoped therefore that preventive strategies designed to reduce chromosome damage will also reduce the rate of cancer in these women. To test for increased mutagenicity of cells from BRCA1 carriers, the frequency of chromosome breaks was measured in cultured blood lymphocytes following in vitro exposure to bleomycin in female BRCA1 carriers and was compared with noncarrier relatives. The frequency of chromosome breaks was also measured in BRCA1 carriers following oral selenium supplementation. Carriers of BRCA1 mutations showed significantly greater mean frequencies of induced chromosome breaks per cell than did healthy noncarrier relatives (0.58 versus 0.39; P < 10(-4)). The frequency of chromosome breaks was greatly reduced following 1 to 3 months of oral selenium supplementation (mean, 0.63 breaks per cell versus 0.40; P < 10(-10)). The mean level of chromosome breaks in carriers following supplementation was similar to that of the noncarrier controls (0.40 versus 0.39). Oral selenium is a good candidate for chemoprevention in women who carry a mutation in the BRCA1 gene. PMID- 15894691 TI - Body mass index in relation to ovarian cancer survival. AB - Evidence for an association between indicators of adiposity and survival after ovarian cancer has been inconsistent. A prospective cohort study was conducted in China to examine the relationship between ovarian cancer survival and body mass index (BMI). From the 214 patients recruited in 1999 to 2000 with histopathologically confirmed invasive epithelial ovarian cancer, 207 patients or their close relatives (96.7% of cases) were traced and followed to 2003. Deaths were recorded and Cox proportional hazards regression was used to obtain hazard ratios (HR) and 95% confidence intervals (95% CI) from multivariate models. Reduced survival was observed among patients with BMI > or = 25 kg/m(2) at 5 years before diagnosis (P = 0.001). There were 98 (59.8%) of 164 patients with BMI <25 kg/m(2) survived to the time of interview compared with only 15 women (34.9%) among the 43 patients whose BMI was > or =25 kg/m(2). The HRs significantly increased with higher BMI at 5 years before diagnosis but not at diagnosis nor at age 21 years. The adjusted HR was 2.33 (95% CI, 1.12-4.87) for BMI of > or =25 versus <20 kg/m(2), with a significant dose-response relationship. The HR was 3.31 (95% CI, 1.26-8.73) among patients who had been overweight or obese at age 21 years, but a linear dose-response was not found. We conclude that premorbid BMI may have independent prognostic significance in ovarian cancer. PMID- 15894692 TI - Semiquantitative human papillomavirus type 16 viral load and the prospective risk of cervical precancer and cancer. AB - We examined whether higher human papillomavirus type 16 (HPV16) viral load predicted risk of cervical intraepithelial neoplasia 3 (CIN3) or cancer (together termed > or =CIN3) within a cohort of 20,810 women followed for 10 years with cytologic screening. Semiquantitative viral load for HPV16 was measured on baseline cervicovaginal specimens using a type-specific hybridization probe test with signal amplification. An increased risk of > or =CIN3 associated with higher HPV16 viral load was found only among cytologically negative women in early follow-up, suggesting that these cases were related to the detection of prevalent lesions missed at baseline. Women with higher HPV16 viral load were more likely to undergo ablative treatment during follow-up than those with lower viral load (P(trend) = 0.008), possibly diminishing any additional risk for > or =CIN3 attributable to higher HPV16 viral loads. PMID- 15894693 TI - Aspirin use and risk of biliary tract cancer: a population-based study in Shanghai, China. AB - The association of gallbladder and bile duct cancers with gallstones, cholecystitis, and cholangitis suggest that chronic inflammation contributes to the carcinogenic process. However, the effect of nonsteroidal anti-inflammatory drugs, such as aspirin, on biliary tract cancer has not been well studied. In a population-based case-control study conducted in Shanghai, China, we examined the relationship between aspirin use and the risk of biliary disease. A total of 627 patients with biliary tract cancer, including cancers of the gallbladder (n = 368), extrahepatic bile duct (n = 191), and ampulla of Vater (n = 68); 1,037 patients with biliary stones; and 958 healthy adults were included in the study. Self-reported data on aspirin use was collected from study participants by in person interview. The prevalence of aspirin use was low, with 5.7% of the population controls being regular users. After controlling for age, sex, education, and biliary stone status, aspirin use was associated with a reduced risk of gallbladder cancer [odds ratio (OR), 0.37; 95% confidence interval (CI), 0.17-0.88]. An inverse relationship was also observed for frequency and duration of use and with younger age when starting use. In addition, there was a nonsignificant reduction in the risk of bile duct (OR, 0.48; 95% CI, 0.19-1.19) and ampullary cancers (OR, 0.22; 95% CI, 0.03-1.65) associated with aspirin use, whereas no clear association was seen with biliary stones (OR, 0.92; 95% CI, 0.59 1.44). Further studies of biliary tract cancer in other populations are needed to confirm these results and to elucidate the mechanisms that underlie the reduced risk associated with use of aspirin and possibly other nonsteroidal anti inflammatory drugs. PMID- 15894694 TI - Association of an exon 1 polymorphism in the IGFBP3 gene with circulating IGFBP-3 levels and colorectal cancer risk: the multiethnic cohort study. AB - Laboratory and seroepidemiologic studies have suggested that insulin-like growth factor binding protein-3 (IGFBP-3), the main binding protein for IGF-I, may be protective against colorectal cancer. We investigated the association of two polymorphisms (A-202C and G2133C) in the IGFBP3 gene with plasma IGF hormone levels among 887 randomly selected participants in the Multiethnic Cohort study. We found that these two genetic variants were in strong linkage disequilibrium and were both inversely associated with plasma IGFBP-3. However, the effect on plasma IGFBP-3 levels was stronger for the G2133C variant than the A-202C variant. Thus, we assessed the colorectal cancer risk associated with the G2133C in a case-control study of 817 cases and 1,995 controls nested within the Multiethnic Cohort study. Under the assumption of dominant genetic model, carriers of the 1233C allele were at 32% increased risk of colorectal cancer [95 % confidence interval (95% CI) for the odds ratio (OR), 1.07-1.62] and that this effect seemed stronger for the rectum (OR for the C allele, 1.95; 95% CI, 1.35 2.83) than the colon (OR, 1.16; 95% CI, 0.92-1.45). These data suggest that the exon 1 G2133C missense variant in IGFBP3 may be a susceptibility factor for colorectal cancer. PMID- 15894695 TI - Polymorphism of the CD30 promoter microsatellite repressive element is associated with development of primary cutaneous lymphoproliferative disorders. AB - Lymphomatoid papulosis is a preneoplastic cutaneous lymphoproliferative disorder characterized by overexpression of CD30, a member of the tumor necrosis factor receptor superfamily. CD30 signaling is known to have an effect on the growth and survival of lymphoid cells. Therefore, we hypothesized that the development of lymphomatoid papulosis and progression to an associated neoplasm such as cutaneous and systemic anaplastic large cell lymphoma may reflect an underlying genetic defect. In this study, we determined that two allelic forms of the CD30 promoter microsatellite repressive element, designated 30M377 and 30M362, are associated with the development of lymphomatoid papulosis and CD30+ lymphomas in lymphomatoid papulosis patients, respectively. These findings suggest that allele specific differences in the control of CD30 transcription may determine the pathogenesis of the spectrum of CD30+ cutaneous lymphoproliferative disorders. PMID- 15894696 TI - Screening for deleterious nonsynonymous single-nucleotide polymorphisms in genes involved in steroid hormone metabolism and response. AB - To facilitate selection of single-nucleotide polymorphisms (SNP) for molecular epidemiologic studies investigating the hormonal carcinogenesis hypothesis, we used two sequence homology-based tools [Sort Intolerant from Tolerant (SIFT) and Polymorphism Phenotype (PolyPhen)] to predict the potential impact a nonsynonymous SNP (nsSNP), which results in an amino acid substitution, may have on the activity of proteins encoded by genes involved in the steroid hormone metabolism and response pathway. We screened 137 variants. Of these, 28% were predicted by SIFT and PolyPhen as having a potentially damaging effect on protein function. Investigation into the association of these variant alleles with hormone-related cancers may prove to be fruitful. PMID- 15894697 TI - Response rates to a mailed survey targeting childhood cancer survivors: a comparison of conditional versus unconditional incentives. AB - OBJECTIVE: Mailed surveys are widely used to collect epidemiologic and health service data. Given that nonresponse can threaten the validity of surveys, modest incentives are often used to increase response rates. A study was undertaken among childhood cancer survivors and their parents to determine if response rate to a mailed survey differed with provision of immediate versus delayed incentives. DESIGN: A self-administered survey designed to ascertain health behaviors was mailed to 397 childhood cancer survivors (and their parents if the survivor was <18 years of age). Subjects were randomized into two groups based on gender, age, race, and cancer type. One group received a 10 US dollars incentive with their blank survey (unconditional incentive), whereas the other group received the incentive upon receipt of their completed survey (conditional incentive). If children were minors, both the parent and the child received incentives. RESULTS: No significant differences in response rates were observed with respect to gender, age, race, or cancer type. However, significant differences in response rates were observed between incentive groups, with unconditional incentives yielding significantly higher response rates than conditional incentives for child survivors who were > or =18 years (64.4% versus 49.0%), as well as younger child survivors (62.5% versus 43.6%) and their parents (64.8% versus 41.5%; all P < 0.05). CONCLUSIONS: The provision of an immediate incentive generated significantly higher response rates to this mailed health survey among childhood cancer survivors and their parents. Given that survey studies are commonly conducted across various pediatric populations, these findings may help inform the design of future pediatric survey research. PMID- 15894698 TI - Association of vegetable intake with urinary 6-sulfatoxymelatonin level. AB - Melatonin is present in plants consumed as vegetables; however, only a limited number of vegetables have been tested for melatonin. The antiproliferative, antioxidative, and immunostimulatory effects of melatonin have been reported from laboratory studies. The potential protective effects of vegetable against cancer and cardiovascular disease may be partially attributable to an increased melatonin intake from vegetables. As a first step to test this hypothesis, we evaluated whether vegetable intake is associated with an increased urinary melatonin in 289 community-dwelling Japanese women. Diet, including vegetable consumption, was assessed with a validated 169-item semiquantitative food frequency questionnaire. Urinary 6-sulfatoxymelatonin (aMT6-s) was measured in the first-void morning urines. There was a significant positive association between vegetable intake and urinary aMT6-s levels. The mean urinary aMT6-s was 16% higher in women with the highest quartile of vegetable intake than it was in those with the lowest quartile of intake. This association may be explained by the melatonin contained in vegetables. However, data should be regarded as preliminary because it is impossible to estimate dietary melatonin intake from vegetables and or from the entire diet because of incomplete data for melatonin in plants. PMID- 15894699 TI - Prenatal and postnatal milk supplementation and adult insulin-like growth factor I: long-term follow-up of a randomized controlled trial. AB - OBJECTIVE: Insulin-like growth factors (IGF) are increasingly recognized as important determinants of adult health, in particular risk of certain cancers. However, little is known about the determinants of adult IGFs and to what degree they may be programmed by early life influences. DESIGN: Randomized controlled trial of prenatal and postnatal milk supplementation among 951 subjects born in 1972 to 1974 in South Wales. MAIN OUTCOME MEASURE: Measures of IGF-I, IGF binding protein 3, and the molar ratio. RESULTS: Data on adult IGFs were available from 663 subjects at a mean age of 25 years. Subjects in the intervention arm had lower IGF-I (-8.5 ng/mL; 95% confidence interval, -15.1 to -1.8, P = 0.01) and ratio (-1.20; 95% confidence interval, -2.33 to -0.04, P = 0.04). These differences could not be explained by follow-up bias or confounding factors. CONCLUSIONS: These results provide experimental data on the role of early life programming either in the intrauterine or postnatal period that may have long term influences on the IGF axis, with potential implications for disease risk. PMID- 15894700 TI - Transition metals in personal samples of PM2.5 and oxidative stress in human volunteers. AB - Ambient particulate matter (PM) has been associated with increased risk of lung cancer. One proposed mechanism is that PM induces oxidative stress mediated by transition metals contained within this mixture. We examined the relationship between the personal exposure to water-soluble transition metals in PM(2.5) and oxidative DNA damage. In 49 students from central Copenhagen, we determined PM(2.5) exposure by personal sampling twice in 1 year, and measured in these PM(2.5) samples the concentration of the soluble transition metals vanadium, chromium, iron, nickel, copper, and platinum. Collected lymphocytes and 24-hour urine samples were analyzed for DNA damage in terms of 7-hydro-8-oxo-2' deoxyguanosine (8-oxodG). We found that the 8-oxodG concentration in lymphocytes was significantly associated with the vanadium and chromium concentrations with a 1.9% increase in 8-oxodG per 1 microg/L increase in the vanadium concentration and a 2.2% increase in 8-oxodG per 1 microg/L increase in the chromium concentration. We have previously reported that in this study population the personal exposure to PM(2.5) was associated with an increase in 8-oxodG in lymphocytes. However, vanadium and chromium were associated with the 8-oxodG concentration in lymphocytes independently of the PM(2.5) mass concentration. The four other transition metals were not associated with 8-oxodG in lymphocytes and none of the transition metals was significantly associated with 8-oxodG in urine. Our results could indicate that vanadium and chromium present in PM(2.5) have an effect on oxidative DNA damage that is independent of particle mass and/or other possible toxic compounds contained within this particulate mixture. PMID- 15894701 TI - Dietary feeding of silibinin prevents early biomarkers of UVB radiation-induced carcinogenesis in SKH-1 hairless mouse epidermis. AB - Solar radiation is the causal etiologic factor in the development of nonmelanoma skin cancer (NMSC). Depletion of the stratospheric ozone layer leads to an increase in ambient UV radiation loads, which are expected to further raise skin cancer incidence in many temperate parts of the world, including the United States, suggesting that skin cancer chemopreventive approaches via biomarker efficacy studies or vice versa are highly warranted. Based on our recent study reporting strong efficacy of silibinin against photocarcinogenesis, we assessed here the protective effects of its dietary feeding on UVB-induced biomarkers involved in NMSC providing a mechanistic rationale for an early-on silibinin efficacy in skin cancer prevention. Dietary feeding of silibinin at 1% dose (w/w) to SKH-1 hairless mice for 2 weeks before a single UVB irradiation at 180 mJ/cm(2) dose resulted in a strong and significant (P < 0.001) decrease in UVB induced thymine dimer-positive cells and proliferating cell nuclear antigen, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling, and apoptotic sunburn cells together with an increase (P < 0.001) in p53 and p21/cip1 positive cell population in epidermis. These findings suggest that dietary feeding of silibinin affords strong protection against UVB-induced damages in skin epidermis by (a) either preventing DNA damage or enhancing repair, (b) reducing UVB-induced hyperproliferative response, and (c) inhibiting UVB-caused apoptosis and sunburn cell formation, possibly via silibinin-caused up-regulation of p53 and p21/cip1 as major UVB-damage control sensors. PMID- 15894702 TI - Microsomal epoxide hydrolase polymorphisms are not associated with colon cancer risk. PMID- 15894703 TI - No association between BRCA2 N372H and breast cancer risk. PMID- 15894704 TI - Measuring DNA damage among smokers. PMID- 15894705 TI - The impact of medical interpreter services on the quality of health care: a systematic review. AB - Twenty-one million Americans are limited in English proficiency (LEP), but little is known about the effect of medical interpreter services on health care quality. Asystematic literature review was conducted on the impact of interpreter services on quality of care. Five database searches yielded 2,640 citations and a final database of 36 articles, after applying exclusion criteria. Multiple studies document that quality of care is compromised when LEP patients need but do not get interpreters. LEP patients' quality of care is inferior, and more interpreter errors occur with untrained ad hoc interpreters. Inadequate interpreter services can have serious consequences for patients with mental disorders. Trained professional interpreters and bilingual health care providers positively affect LEP patients' satisfaction, quality of care, and outcomes. Evidence suggests that optimal communication, patient satisfaction, and outcomes and the fewest interpreter errors occur when LEP patients have access to trained professional interpreters or bilingual providers. PMID- 15894706 TI - Access to ambulatory medical and long-term care services among elderly Medicare and Medicaid beneficiaries: organizational, financial, and geographic barriers. AB - The comprehensive insurance coverage afforded low-income elders with both Medicare and Medicaid coverage (dual enrollees) has substantially reduced financial barriers to care. However, other studies show reduced and less appropriate utilization patterns among dual enrollees compared to Medicare beneficiaries with private supplemental insurance, suggesting access barriers remain. This study found that 59 percent of elderly dual enrollees needed an ambulatory medical or long-term care service in a 1-year period. One third of these individuals experienced barriers to access; organizational and geographic barriers were more prevalent than financial barriers. African American race, trouble paying basic living expenses, fair or poor health status, and an unfavorable assessment of physician information giving were significantly associated with an increased likelihood of organizational and geographic access barriers among elderly dual enrollees. PMID- 15894707 TI - The impact of managed care on publicly funded outpatient adolescent substance abuse treatment: service use and six-month outcomes in Oregon and Washington. AB - This study assessed the impact of managed care on publicly funded adolescent substance abuse treatment by comparing differences in service utilization and outcomes across prospective samples from two states: Oregon, which uses managed care practices in service financing and delivery, and Washington, which does not. One hundred and six adolescents from Washington and 94 from Oregon, who entered outpatient substance abuse treatment in 1998 and 1999, completed self-report surveys about their substance use before and after receiving treatment (follow-up rate = 75 percent). In addition, clinical chart reviews conducted at the 6-month follow-up assessed the type and amount of treatment these adolescents received during the study period. It was found that service utilization and treatment outcomes were comparable across the two state samples. The evidence presented here suggests that managed care is capable of delivering substance abuse treatment services of comparable quality to state-administered substance abuse treatment services. PMID- 15894708 TI - Explaining variation in physician practice patterns and their propensities to recommend services. AB - Variations in physician practice patterns have important implications for quality and cost. The purpose of this article is to explain variation in physicians' practice patterns in terms of physician personal characteristics, practice setting, patient population, and managed care involvement. Data on 2,455 primary care physicians were derived from the Community Tracking Study Physician Survey (1996-1997). Factor scores were determined based on responses to three clinical scenarios that represent discretionary medical decisions. These scenarios include a specialist referral for benign prostatic hyperplasia, prescription drugs for elevated cholesterol, and an office visit for vaginal discharge. Physician age, being a foreign medical school graduate, being a solo practitioner, and having a larger proportion of Medicaid patients were all associated with higher factor scores, a greater likelihood of ordering a service. Being board certified was associated with lower factor scores. Managed care involvement was not a significant predictor of factor scores. PMID- 15894709 TI - It isn't just about choice: the potential of a public performance report to affect the public image of hospitals. AB - Can a well-designed public performance report affect the public image of hospitals? Using a pre/postdesign and telephone interviews, consumer views and reports of their use of public hospital report are examined. The findings show that the report did influence consumer views about the quality of individual hospitals in the community 2 to 4 months after the release of the report. PMID- 15894711 TI - A novel germline mutation, 1793delG, of the MEN1 gene underlying multiple endocrine neoplasia type 1. AB - Pulmonary carcinoids are rare neuroendocrine tumors which comprise 1-2% of all lung tumors. They usually occur sporadically; however, their association with multiple endocrine neoplasia type 1 (MEN1) syndrome has been documented. We report a case of a Thai woman with a pulmonary carcinoid tumor and a null cell pituitary tumor. Her family history was unremarkable for any MEN-related lesions. Genetic testing revealed a novel deletion mutation at exon 10 (1793delG) of the MEN1 gene, resulting in a stop codon 26 amino acids downstream. This mutation is predicted to cause a loss of the second nuclear localization signal of the menin protein. PMID- 15894712 TI - Arsenite-induced aryl hydrocarbon receptor nuclear translocation results in additive induction of phase I genes and synergistic induction of phase II genes. AB - Complex mixtures of carcinogenic metalloids, such as arsenic, and polycyclic aromatic hydrocarbons or halogenated aromatic hydrocarbons are common environmental contaminants. The biological consequences of exposure to these mixtures are unpredictable and, although the health effects of individual chemicals may be known, the toxicity of environmental mixtures is largely unexplored. Arsenic, not a potent mutagen by itself, is co-mutagenic with many DNA-damaging agents. Mixtures of arsenite plus benzo[a]pyrene (B[a]P) augment B[a]P mutagenicity, suggesting that arsenite might uncouple expression of phase I and II genes responsible for detoxification. We have studied the effects of arsenite exposure on the activation of the aryl hydrocarbon receptor (AHR) and its subsequent role in gene transactivation. Treatment of mouse Hepa-1 cells with arsenite induces AHR nuclear translocation and binding to the Cyp1a1 gene promoter with the same efficiency as tetrachlorodibenzo-p-dioxin (TCDD), the most potent ligand of the AHR; however, TCDD and B[a]P are an order of magnitude more potent than arsenite in up-regulating Cyp1a1 transcription. Global profiling analyses of cells treated with arsenite plus B[a]P indicate that several phase I and II detoxification genes are in some cases additively and in others synergistically deregulated by the mixtures. Real-time reverse transcription polymerase chain reaction analyses of mouse embryonic fibroblasts showed that the mixtures had an additive effect on the mRNA levels of Cyp1b1, a prototypical phase I detoxification gene, and an AHR-dependent synergistic effect on the corresponding levels of Nqo1, a prototypical phase II gene. We conclude that exposure to arsenite/B[a]P mixtures causes regulatory changes in the expression of detoxification genes that ultimately affect the metabolic activation and disposition of toxicants. PMID- 15894713 TI - Single nucleotide polymorphisms in the human norepinephrine transporter gene affect expression, trafficking, antidepressant interaction, and protein kinase C regulation. AB - The role of norepinephrine (NE) in attention, memory, affect, stress, heart rate, and blood pressure implicates NE in psychiatric and cardiovascular disease. The norepinephrine transporter (NET) mediates reuptake of released catecholamines, thus playing a role in the limitation of signaling strength in the central and peripheral nervous systems. Nonsynonymous single nucleotide polymorphisms (SNPs) in the human NET (hNET) gene that influence transporter function can contribute to disease, such as the nonfunctional transporter, A457P, identified in orthostatic intolerance. Here, we examine additional amino acid variants that have been identified but not characterized in populations that include cardiovascular phenotypes. Variant hNETs were expressed in COS-7 cells and were assayed for protein expression and trafficking using cell-surface biotinylation and Western blot analysis, transport of radiolabeled substrate, antagonist interaction, and regulation through protein kinase C (PKC)-linked pathways by the phorbol ester beta-phorbol-12-myristate-13-acetate. We observed functional perturbations in 6 of the 10 mutants studied. Several variants were defective in trafficking and transport, with the most dramatic effect observed for A369P, which was completely devoid of the fully glycosylated form of transporter protein, was retained intracellularly, and lacked any transport activity. Furthermore, A369P and another trafficking variant, N292T, impeded surface expression of hNET when coexpressed. F528C demonstrated increased transport and, remarkably, exhibited both insensitivity to down-regulation by PKC and a decrease in potency for the tricyclic antidepressant desipramine. These findings reveal functional deficits that are likely to compromise NE signaling in SNP carriers in the population and identify key regions of NET contributing to transporter biosynthesis, activity, and regulation. PMID- 15894714 TI - Pigment deficiency in nightshade/tobacco cybrids is caused by the failure to edit the plastid ATPase alpha-subunit mRNA. AB - The subgenomes of the plant cell, the nuclear genome, the plastome, and the chondriome are known to interact through various types of coevolving macromolecules. The combination of the organellar genome from one species with the nuclear genome of another species often leads to plants with deleterious phenotypes, demonstrating that plant subgenomes coevolve. The molecular mechanisms behind this nuclear-organellar incompatibility have been elusive, even though the phenomenon is widespread and has been known for >70 years. Here, we show by direct and reverse genetic approaches that the albino phenotype of a flowering plant with the nuclear genome of Atropa belladonna (deadly nightshade) and the plastome of Nicotiana tabacum (tobacco) develops as a result of a defect in RNA editing of a tobacco-specific editing site in the plastid ATPase alpha subunit transcript. A plastome-wide analysis of RNA editing in these cytoplasmic hybrids and in plants with a tobacco nucleus and nightshade chloroplasts revealed additional defects in the editing of species-specific editing sites, suggesting that differences in RNA editing patterns in general contribute to the pigment deficiencies observed in interspecific nuclear-plastidial incompatibilities. PMID- 15894715 TI - Differential recognition of highly divergent downy mildew avirulence gene alleles by RPP1 resistance genes from two Arabidopsis lines. AB - The perception of downy mildew avirulence (Arabidopsis thaliana Recognized [ATR]) gene products by matching Arabidopsis thaliana resistance (Recognition of Peronospora parasitica [RPP]) gene products triggers localized cell death (a hypersensitive response) in the host plant, and this inhibits pathogen development. The oomycete pathogen, therefore, is under selection pressure to alter the form of these gene products to prevent detection. That the pathogen maintains these genes indicates that they play a positive role in pathogen survival. Despite significant progress in cloning plant RPP genes and characterizing essential plant components of resistance signaling pathways, little progress has been made in identifying the oomycete molecules that trigger them. Concluding a map-based cloning effort, we have identified an avirulence gene, ATR1NdWsB, that is detected by RPP1 from the Arabidopsis accession Niederzenz in the cytoplasm of host plant cells. We report the cloning of six highly divergent alleles of ATR1NdWsB from eight downy mildew isolates and demonstrate that the ATR1NdWsB alleles are differentially recognized by RPP1 genes from two Arabidopsis accessions (Niederzenz and Wassilewskija). RPP1-Nd recognizes a single allele of ATR1NdWsB; RPP1-WsB also detects this allele plus three additional alleles with divergent sequences. The Emco5 isolate expresses an allele of ATR1NdWsB that is recognized by RPP1-WsB, but the isolate evades detection in planta. Although the Cala2 isolate is recognized by RPP1-WsA, the ATR1NdWsB allele from Cala2 is not, demonstrating that RPP1-WsA detects a novel ATR gene product. Cloning of ATR1NdWsB has highlighted the presence of a highly conserved novel amino acid motif in avirulence proteins from three different oomycetes. The presence of the motif in additional secreted proteins from plant pathogenic oomycetes and its similarity to a host-targeting signal from malaria parasites suggest a conserved role in pathogenicity. PMID- 15894716 TI - Phylloplanins of tobacco are defensive proteins deployed on aerial surfaces by short glandular trichomes. AB - In plants, defensive proteins secreted to leaf aerial surfaces have not previously been considered to be a strategy of pathogen resistance, and the general occurrence of leaf surface proteins is not generally recognized. We found that leaf water washes (LWW) of the experimental plant Nicotiana tabacum tobacco introduction (TI) 1068 contained highly hydrophobic, basic proteins that inhibited spore germination and leaf infection by the oomycete pathogen Peronospora tabacina. We termed these surface-localized proteins tobacco phylloplanins, and we isolated the novel gene T-Phylloplanin (for Tobacco Phylloplanin) and its promoter from N. tabacum. Escherichia coli-expressed T phylloplanin inhibited P. tabacina spore germination and greatly reduced leaf infection. The T-phylloplanin promoter, when fused to the reporter genes beta glucuronidase and green fluorescent protein, directed biosynthesis only in apical tip cell clusters of short, procumbent glandular trichomes. Here, we provide evidence for a protein-based surface defense system in the plant kingdom, wherein protein biosynthesis in short, procumbent glandular trichomes allows surface secretion and deposition of defensive phylloplanins on aerial surfaces as a first point-of-contact deterrent to pathogen establishment. As yet uncharacterized surface proteins have been detected on most plant species examined. PMID- 15894717 TI - Identification and functional analysis of in vivo phosphorylation sites of the Arabidopsis BRASSINOSTEROID-INSENSITIVE1 receptor kinase. AB - Brassinosteroids (BRs) regulate multiple aspects of plant growth and development and require an active BRASSINOSTEROID-INSENSITIVE1 (BRI1) and BRI1-ASSOCIATED RECEPTOR KINASE1 (BAK1) for hormone perception and signal transduction. Many animal receptor kinases exhibit ligand-dependent oligomerization followed by autophosphorylation and activation of the intracellular kinase domain. To determine if early events in BR signaling share this mechanism, we used coimmunoprecipitation of epitope-tagged proteins to show that in vivo association of BRI1 and BAK1 was affected by endogenous and exogenous BR levels and that phosphorylation of both BRI1 and BAK1 on Thr residues was BR dependent. Immunoprecipitation of epitope-tagged BRI1 from Arabidopsis thaliana followed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) identified S-838, S 858, T-872, and T-880 in the juxtamembrane region, T-982 in the kinase domain, and S-1168 in C-terminal region as in vivo phosphorylation sites of BRI1. MS analysis also strongly suggested that an additional two residues in the juxtamembrane region and three sites in the activation loop of kinase subdomain VII/VIII were phosphorylated in vivo. We also identified four specific BAK1 autophosphorylation sites in vitro using LC/MS/MS. Site-directed mutagenesis of identified and predicted BRI1 phosphorylation sites revealed that the highly conserved activation loop residue T-1049 and either S-1044 or T-1045 were essential for kinase function in vitro and normal BRI1 signaling in planta. Mutations in the juxtamembrane or C-terminal regions had only small observable effects on autophosphorylation and in planta signaling but dramatically affected phosphorylation of a peptide substrate in vitro. These findings are consistent with many aspects of the animal receptor kinase model in which ligand-dependent autophosphorylation of the activation loop generates a functional kinase, whereas phosphorylation of noncatalytic intracellular domains is required for recognition and/or phosphorylation of downstream substrates. PMID- 15894718 TI - Rapid elimination of cefaclor from the cerebrospinal fluid is mediated by a benzylpenicillin-sensitive mechanism distinct from organic anion transporter 3. AB - The purpose of this study was to investigate the carrier-mediated elimination of cephalosporins from the cerebrospinal fluid (CSF) via the choroid plexus. Cefaclor and cefalexin are structural analogs with similar lipophilicity, differing by only one functional group (cefaclor, -Cl; cephalexin, -CH(3)), and they are substrates of rat peptide transporter PEPT2 with similar transport activities. However, cefaclor was cleared from the CSF more rapidly than cefalexin after intracerebroventricular administration (the elimination rate constants were 0.11 and 0.050 min(-1), respectively). The elimination of cefaclor from the CSF was inhibited by benzylpenicillin, but not by glycylsarcosine (GlySar), whereas GlySar, but not benzylpenicillin, had an inhibitory effect on the elimination of cefalexin from the CSF. The uptake of cefaclor by the freshly isolated rat choroid plexus was saturable, with a K(m) value of 250 muM, and the uptake clearance corresponding to saturable components accounts for the major part of the in vivo clearance from the CSF (17 versus 26 mul/min, respectively). The uptake of cefaclor by the choroid plexus was inhibited by benzylpenicillin, but not by GlySar. However, the inhibitory effect of benzylpenicillin was weaker than expected from its own K(m) value, and furthermore, organic anion transporter (Oat)3 substrates (cimetidine or p-aminohippurate) had no effect. These results suggest that cefaclor and cefalexin are eliminated from the CSF by different transporters, and rapid elimination of cefaclor from the CSF is accounted for by a benzylpenicillin-sensitive mechanism distinct from Oat3. A slight modification of a single chemical group of cephalosporins can greatly affect the contribution of the transporters involved, and their duration in the CSF. PMID- 15894719 TI - Vet to be struck off after charges found proven. PMID- 15894724 TI - Dealing with the aftermath: the role of the vet following conflict or disaster. PMID- 15894727 TI - Preliminary study using an indirect ELISA for the detection of serum antibodies to Alternaria in domestic cats. AB - Alternaria is a saprophytic fungus that is widespread in the environment; it is an opportunistic pathogen and causes disease in human beings and domestic animals. Fungal spores gain entry to the host through skin lesions and cause slow growing, soft to firm, subcutaneous swellings, either with or without ulcers. An indirect ELISA was developed for the detection of anti-Alternaria immunoglobulin G (IgG) antibodies in serum to determine the prevalence of Alternaria exposure in domestic cats. Fifty-two of 63 cats had detectable levels of anti-Alternaria IgG antibody. There were no correlations between the concentration of antibody and the sex, breed or living environment of the cats, but cats less than two years of age had significantly lower concentrations than older cats. The cats with disease caused by culture-confirmed Alternaria infections did not have significantly higher concentrations of antibody than the healthy cats or cats with other diseases. PMID- 15894728 TI - Biochemical reference ranges for groups of ewes of different ages. AB - Reference biochemical ranges for young adult and adult ewes in mid-gestation were derived from 83 pooled serum samples taken from 30 flocks in the Province of Quebec, Canada. In each flock the samples were pooled into three age categories and each category contained between six and 10 samples. The blood samples were taken by jugular venepuncture in December 1999. All the values were within the normal published ranges. As the age of the ewes increased there were slight increases in the concentrations of globulin and total protein and decreases in the concentrations of calcium, phosphorus, creatinine, glucose and albumin, and in the activities of alkaline phosphatase and glutathione peroxidase. PMID- 15894729 TI - Evaluation of the analgesic effect of lidocaine and bupivacaine used to provide a brachial plexus block for forelimb surgery in 10 dogs. AB - Twenty adult dogs weighing between 1.4 and 53.5 kg and aged between six months and nine years were anaesthetised and the brachial plexus was localised with the aid of a nerve stimulator. In 10 of the dogs a brachial plexus block was induced with a mixture of lidocaine and bupivacaine and the other 10 each received 0.25 ml/kg saline as a control. The end-tidal isoflurane concentration was maintained between 1.3 and 1.4 per cent during surgery for carpal arthrodesis or a fracture of the radius or ulna. Acute heart rate or blood pressure increases of 20 per cent or more were treated with 1 microg/kg fentanyl intravenously. Postoperatively, signs of pain were scored by a single blinded observer at hourly intervals until eight hours after the block had been induced, on a scale from 0 to 18. Dogs with pain scores above 5 received 0.1 to 0.2 mg/kg methadone intravenously, repeated as necessary. During surgery the control dogs received significantly more fentanyl (median 0.05 microg/kg/minute, range 0.02 to 0.20 microg/kg/minute) than the group given local anaesthetic (median 0 microg/kg/minute, range 0 to 0.02 microg/kg/minute). Postoperatively, the control group required significantly more methadone (median 0.2 mg/kg, range 0.1 to 1 mg/kg) than the treated group (median 0 mg/kg, range 0 to 0.13 mg/kg). PMID- 15894730 TI - Masseter myodegeneration as a cause of trismus or dysphagia in adult horses. AB - The medical records of eight horses with histological evidence of myodegeneration of the masseter muscles were examined. While they were alive their most common clinical signs had included difficulty in eating or opening their mouths, weight loss, difficulty in moving, and noticeable atrophy of the masseter muscles. The serum activities of muscle enzymes were abnormally high in all of the horses. Whole blood and/or liver selenium and vitamin E concentrations were less than the reference ranges in some of the horses. The lesions varied with the stage of the disease and consisted of swelling and discoloration, or muscle atrophy and fibrosis. Histologically, the muscle changes ranged from acute to subacute degeneration, with regenerative changes accompanying ongoing degeneration, to chronic degeneration with fibrotic replacement of muscle tissue. There were changes in the masseter muscle of all the horses, but some had widespread lesions in skeletal muscle, and a few also had myocardial lesions. PMID- 15894731 TI - Efficacy of live infectious bronchitis vaccines against a novel European genotype, Italy 02. PMID- 15894732 TI - Use of ultrafast Papanicolaou stain for exfoliative vaginal cytology in bitches. PMID- 15894733 TI - Spontaneous squamous cell carcinoma of the cheek pouch in two dwarf hamsters (Phodopus sungorus). PMID- 15894734 TI - Extracranial meningioma in a dairy cow. PMID- 15894736 TI - RCVS Practice Standards Scheme. PMID- 15894735 TI - Residues surveillance of OTMS animals. PMID- 15894737 TI - Shower dipping in diazinon or cypermethrin dipwash to control ovine psoroptic mange (sheep scab). PMID- 15894739 TI - Erysipelas outbreak in racing pigeons following ingestion of compost. PMID- 15894740 TI - Garden bird health. PMID- 15894741 TI - Genome-wide identification of genes expressed in Arabidopsis pistils specifically along the path of pollen tube growth. AB - Plant reproductive development is dependent on successful pollen-pistil interactions. In crucifers, the pollen tube must breach the stigma surface and burrow through the extracellular matrix of the stigma epidermal cells and transmitting tract cells before reaching its ovule targets. The high degree of specificity in pollen-pistil interactions and the precision of directional pollen tube growth suggest that signals are continually being exchanged between pollen/pollen tubes and cells of the pistil that line their path. However, with few exceptions, little is known about the genes that control these interactions. The specialized functions of stigma epidermal cells and transmitting tract cells are likely to depend on the activity of genes expressed specifically in these cells. In order to identify these genes, we used the Arabidopsis (Arabidopsis thaliana) ATH1 microarray to compare the whole-genome transcriptional profiles of stigmas and ovaries isolated from wild-type Arabidopsis and from transgenic plants in which cells of the stigma epidermis and transmitting tract were specifically ablated by expression of a cellular toxin. Among the 23,000 genes represented on the array, we identified 115 and 34 genes predicted to be expressed specifically in the stigma epidermis and transmitting tract, respectively. Both gene sets were significantly enriched in predicted secreted proteins, including potential signaling components and proteins that might contribute to reinforcing, modifying, or remodeling the structure of the extracellular matrix during pollination. The possible role of these genes in compatible and incompatible pollen-pistil interactions is discussed. PMID- 15894742 TI - Regulation of plant disease resistance, stress responses, cell death, and ethylene signaling in Arabidopsis by the EDR1 protein kinase. AB - ENHANCED DISEASE RESISTANCE 1 (EDR1) encodes a CTR1-like kinase and was previously reported to function as a negative regulator of disease resistance and ethylene-induced senescence. Here, we report that the edr1 mutant displays enhanced stress responses and spontaneous necrotic lesions under drought conditions in the absence of pathogen, suggesting that EDR1 is also involved in stress response signaling and cell death regulation. Double mutant analysis revealed that these drought-induced phenotypes require salicylic acid but not ethylene signaling pathways. In addition, the edr1-mediated ethylene-induced senescence phenotype was suppressed by mutations in EIN2, but not by mutations in SID2, PAD4, EDS1, or NPR1, suggesting that EDR1 functions at a point of cross talk between ethylene and salicylic acid signaling that impinges on senescence and cell death. Two edr1-associated phenotypes, drought-induced growth inhibition and ethylene-induced senescence, were suppressed by mutations in ORE9, implicating ubiquitin-mediated protein degradation in the regulation of these phenotypes. However, the ore9 mutation did not suppress edr1-mediated enhanced disease resistance to powdery mildew or spontaneous lesions, indicating that these phenotypes are controlled by separate signaling pathways. To investigate the function of the EDR1 kinase domain, we expressed the C-terminal third of EDR1 in wild-type Columbia and edr1 backgrounds under the control of a dexamethasone inducible promoter. Overexpression of the EDR1 kinase domain in an edr1 background had no obvious effect on edr1-associated phenotypes. However, overexpression of the EDR1 kinase domain in a wild-type Columbia background caused dominant negative phenotypes, including enhanced disease resistance to powdery mildew and enhanced ethylene-induced senescence; thus, the overexpressed EDR1 kinase domain alone does not exert EDR1 function, but rather negatively affects the function of native EDR1 protein. PMID- 15894743 TI - Analysis of a sugar response mutant of Arabidopsis identified a novel B3 domain protein that functions as an active transcriptional repressor. AB - A recessive mutation hsi2 of Arabidopsis (Arabidopsis thaliana) expressing luciferase (LUC) under control of a short promoter derived from a sweet potato (Ipomoea batatas) sporamin gene (Spo(min)LUC) caused enhanced LUC expression under both low- and high-sugar conditions, which was not due to increased level of abscisic acid. The hsi2 mutant contained a nonsense mutation in a gene encoding a protein with B3 DNA-binding domain. HSI2 and two other Arabidopsis proteins appear to constitute a novel subfamily of B3 domain proteins distinct from ABI3, FUS3, and LEC2, which are transcription activators involved in seed development. The C-terminal part of HSI2 subfamily proteins contained a sequence similar to the ERF-associated amphiphilic repression (EAR) motif. Deletion of the C-terminal portion of HSI2 lost in the hsi2 mutant caused reduced nuclear targeting of HSI2. Null allele of HSI2 showed even higher Spo(min)LUC expression than the hsi2 mutant, whereas overexpression of HSI2 reduced the LUC expression. Transient coexpression of 35SHSI2 with Spo(min)LUC in protoplasts repressed the expression of LUC activity, and deletion or mutation of the EAR motif significantly reduced the repression activity of HSI2. These results indicate that HSI2 and related proteins are B3 domain-EAR motif active transcription repressors. PMID- 15894744 TI - Starch-related alpha-glucan/water dikinase is involved in the cold-induced development of freezing tolerance in Arabidopsis. AB - Cold-induced soluble sugar accumulation enhances the degree of freezing tolerance in various cold-hardy plants including Arabidopsis (Arabidopsis thaliana), where soluble sugars accumulate in only a few hours at 2 degrees C. Hence, along with photosynthesis, starch degradation might play a significant role in cold-induced sugar accumulation and enhanced freezing tolerance. Starch-related alpha glucan/water dikinase (EC 2.7.9.4), encoded by Arabidopsis STARCH EXCESS 1 (SEX1), is hypothesized to regulate starch degradation in plastids by phosphorylating starch, thereby ensuring better accessibility by starch-degrading enzymes. Here, we show that Arabidopsis sex1 mutants, when incubated at 2 degrees C for 1 d, were unable to accumulate maltooligosaccharides or normal glucose and fructose levels. In addition, they displayed impaired freezing tolerance. After 7 d at 2 degrees C, sex1 mutants did not show any of the above abnormal phenotypes but displayed slightly higher leaf starch contents. The impaired freezing tolerance of sex1 mutants was restored by overexpression of wild-type SEX1 cDNA using the cauliflower mosaic virus 35S promoter. The results demonstrate a genetic link between the SEX1 locus and plant freezing tolerance, and show that starch degradation is important for enhanced freezing tolerance during an early phase of cold acclimation. However, induction of starch degradation was not accompanied by significant changes in alpha-glucan/water dikinase activity in leaf extracts and preceded cold-induced augmentation of SEX1 transcripts. Therefore, we conclude that augmentation of SEX1 transcripts might be a homeostatic response to low temperature, and that starch degradation during an early phase of cold acclimation could be regulated by a component(s) of a starch degradation pathway(s) downstream of SEX1. PMID- 15894745 TI - Arabidopsis thickvein mutation affects vein thickness and organ vascularization, and resides in a provascular cell-specific spermine synthase involved in vein definition and in polar auxin transport. AB - Polar auxin transport has been implicated in the induction of vascular tissue and in the definition of vein positions. Leaves treated with chemical inhibitors of polar auxin transport exhibited vascular phenotypes that include increased vein thickness and vascularization. We describe a recessive mutant, thickvein (tkv), which develops thicker veins in leaves and in inflorescence stems. The increased vein thickness is attributable to an increased number of vascular cells. Mutant plants have smaller leaves and shorter inflorescence stems, and this reduction in organ size and height is accompanied by an increase in organ vascularization, which appears to be attributable to an increase in the recruitment of cells into veins. Furthermore, although floral development is normal, auxin transport in the inflorescence stem is significantly reduced in the mutant, suggesting that the defect in auxin transport is responsible for the vascular phenotypes. In the primary root, the veins appear morphologically normal, but root growth in the tkv mutant is hypersensitive to exogenous cytokinin. The tkv mutation was found to reside in the ACL5 gene, which encodes a spermine synthase and whose expression is specific to provascular cells. We propose that ACL5/TKV is involved in vein definition (defining the boundaries between veins and nonvein regions) and in polar auxin transport, and that polyamines are involved in this process. PMID- 15894746 TI - Very late thrombosis after implantation of sirolimus eluting stent. AB - Stent thrombosis after sirolimus eluting stent implantation has been reported to occur at six hours to 375 days after the procedure and usually within the two weeks after discontinuation of antiplatelet medication. A case is reported of very late stent thrombosis after 17 months of sirolimus eluting stent implantation and eight months after clopidogrel discontinuation despite aspirin continuation. This case underlines the possible need for long term antiplatelet medication among patients receiving sirolimus eluting stents. PMID- 15894747 TI - Managing a complication after direct stenting: removal of a maldeployed stent with rotational atherectomy. AB - A 40 year old patient presented with acute anterior wall infarction. A non calcified lesion was stented directly without significant expansion of the stent. Rotational atherectomy successfully removed parts of the maldeployed stent and resistant arterial wall substance allowing full dilatation of the lesion. PMID- 15894748 TI - Molecular technique identifies the pathogen responsible for culture negative infective endocarditis. AB - A case of culture negative endocarditis complicated by immune complex glomerulonephritis and severe aortic regurgitation necessitated aortic valve replacement. Empirical treatment with penicillin and gentamicin according to UK guidelines was started. The pathogen, Streptococcus sanguis, was later identified by polymerase chain reaction amplification and sequencing of bacterial 16S ribosomal RNA. This molecular technique is likely to be of increasing importance in determining the aetiology of culture negative infective endocarditis, thus providing essential treatment and epidemiological information. PMID- 15894749 TI - Complications of supra-annular mitral valve placement in infants. AB - Two infants underwent supra-annular placement of prosthetic mitral valves. The objective of this strategy was to insert a larger valve and delay replacement. This approach was initially successful but by two and three years later the patients developed impairment of cardiac function. The prosthesis decreased the volume and compliance of the left atrium causing high left atrial and pulmonary venous pressures. The "ventricularised" atrium below the prosthesis dilated. In neither case was it possible to delay second valve replacement. PMID- 15894750 TI - Left atrial myxoma in transplanted heart. AB - Primary cardiac tumours are discovered in about 1 in 2000 necropsies. Cardiac myxomas make up half of all benign intracardiac tumours. Seventy five per cent are located in the left atrium. The first left atrial myxoma arising from the left atrium of a transplanted heart is reported. PMID- 15894751 TI - Rheumatic involvement of all four cardiac valves. AB - Rheumatic involvement of all four heart valves is rare. A 35 year old woman presented with gradually progressive exertional dyspnoea for the preceding 10 years. On evaluation she was in atrial fibrillation with congestive heart failure. Clinical examination found evidence of stenosis of the mitral aortic and tricuspid valves with a history of rheumatic fever in childhood. Transthoracic echocardiography showed the involvement of all four cardiac valves. Few reports are available in the literature describing rheumatic quadrivalvar damage. Operator awareness of possible rheumatic involvement of all four valves is essential for appropriate diagnosis. PMID- 15894752 TI - Fatal infection after rapamycin eluting coronary stent implantation. AB - Septic complications after coronary stenting are extremely rare. The occurrence of cardiac related sepsis after rapamycin eluting stent deployment has not been previously reported. The potential role of drug eluting stents in locally blunting the innate response to bacterial agents is discussed. PMID- 15894753 TI - The changing face of acute myocardial infarction. AB - The appointment of new cardiologists and their involvement in emergency care may lead to decreased mortality in patients suffering acute myocardial infarction. PMID- 15894754 TI - A guide catheter removed a massive intracoronary thrombus: a case of acute coronary syndrome. PMID- 15894755 TI - Xanthine oxidase inhibition for chronic heart failure: is allopurinol the next therapeutic advance in heart failure? AB - A substantial body of evidence has accumulated to suggest a role for the xanthine oxidase metabolic pathway in the pathophysiology of chronic heart failure and other cardiovascular diseases. PMID- 15894756 TI - Educational governance for the regulation of industry sponsored continuing medical education in interventional and device based therapies. AB - The relationship between industry and clinicians in educational programmes needs to be regulated. Industry may be best placed to deliver educational programmes in "craft" related specialties and particularly in areas where device implantation/technology based therapy has a major clinical role. The authors supervise industry sponsored clinical teaching at a purpose built independent teaching facility, and have developed the concept of educational governance to regulate their relationship with their industry sponsor and that concept is presented. PMID- 15894757 TI - Pregnancy in women with congenital heart disease: the importance of evaluation and counselling. AB - Issues surrounding the growing population of pregnant women with congenital heart disease are becoming increasingly important as more women with surgically corrected heart conditions reach childbearing age. PMID- 15894758 TI - Prophylaxis of infective endocarditis: French recommendations 2002. PMID- 15894759 TI - Sixteen row cardiac computed tomography in the diagnosis of arrhythmogenic right ventricular cardiomyopathy. PMID- 15894760 TI - Is coronary heart disease rising in India? A systematic review based on ECG defined coronary heart disease. AB - The aim of this study was to investigate whether coronary heart disease (CHD) is rising in India and assess the quality of the evidence. Thirty one studies were reviewed. The sample sizes of the studies varied from approximately 500 to 14,000, with response rates generally over 90%. Many did not fulfil basic requirements for epidemiological research and most studies were in or around Delhi. The three incidence studies used different diagnostic criteria; however, the incidence of myocardial infarction (MI) in urban India in the 14 years to 1991 remained similar at about 6/1000 in males and 2/1000 in females. Prevalence range was higher in urban than rural areas in men (35-90/1000 v 17-45/1000) and women (28-93/1000 v 13-43/1000). There was no clear rise in age specific rates in men over a 27 year period, whereas a rise was seen in women. In conclusion, using a relatively objective measure of CHD it was found that CHD is more common in urban than rural areas of India, but there was little evidence of a rise in CHD over time, especially in men. PMID- 15894761 TI - Reduced between-hospital variation in short term survival after acute myocardial infarction: the result of improved cardiac care? AB - OBJECTIVES: To re-examine interhospital variation in 30 day survival after acute myocardial infarction (AMI) 10 years on to see whether the appointment of new cardiologists and their involvement in emergency care has improved outcome after AMI. DESIGN: Retrospective cohort study. SETTING: Acute hospitals in Scotland. PARTICIPANTS: 61,484 patients with a first AMI over two time periods: 1988-1991; and 1998-2001. MAIN OUTCOME MEASURES: 30 day survival. RESULTS: Between 1988 and 1991, median 30 day survival was 79.2% (interhospital range 72.1-85.1%). The difference between highest and lowest was 13.0 percentage points (age and sex adjusted, 12.1 percentage points). Between 1998 and 2001, median survival rose to 81.6% (and range decreased to 78.0-85.6%) with a difference of 7.6 (adjusted 8.8) percentage points. Admission hospital was an independent predictor of outcome at 30 days during the two time periods (p < 0.001). Over the period 1988-1991, the odds ratio for death ranged, between hospitals, from 0.71 (95% confidence interval (CI) 0.58 to 0.88) to 1.50 (95% CI 1.19 to 1.89) and for the period 1998 2001 from 0.82 (95% CI 0.60 to 1.13) to 1.46 (95% CI 1.07 to 1.99). The adjusted risk of death was significantly higher than average in nine of 26 hospitals between 1988 and 1991 but in only two hospitals between 1998 and 2001. CONCLUSIONS: The average 30 day case fatality rate after admission with an AMI has fallen substantially over the past 10 years in Scotland. Between-hospital variation is also considerably less notable because of better survival in the previously poorly performing hospitals. This suggests that the greater involvement of cardiologists in the management of AMI has paid dividends. PMID- 15894762 TI - A paclitaxel coated stent used for in-stent restenosis within a sirolimus coated stent fails to protect against recurrent restenosis. PMID- 15894763 TI - Post-challenge hyperinsulinaemia rather than hyperglycaemia is associated with the severity of coronary artery disease in patients without a previous diagnosis of diabetes mellitus. AB - OBJECTIVE: To ascertain the prevalence of abnormal glucose metabolism in patients with coronary artery disease (CAD) but no previous diagnosis of diabetes mellitus (DM) and to examine the relation between the severity of CAD and responses of glucose and insulin to the glucose tolerance test. METHODS AND RESULTS: Abnormalities of glucose metabolism and insulin response were analysed in 144 patients with CAD without a previous diagnosis of DM who underwent both coronary arteriography and 75 g oral glucose tolerance test. The proportions of impaired and diabetic glucose tolerance were very high (39% for impaired and 21% for diabetic glucose tolerance); only 40% had normal glucose tolerance. The parameters of glucose metabolism were not associated with the number of diseased coronary arteries or the presence of previous myocardial infarction (MI). However, the insulin concentration at 60 minutes or 120 minutes after glucose challenge, insulin area, and the ratio of insulin to glucose area were significantly higher in patients with significant coronary stenosis and with previous MI. Fasting glucose concentration and most conventional risk factors did not predict post-challenge hyperinsulinaemia. CONCLUSION: Patients with CAD without a previous diagnosis of DM had a high prevalence of abnormal glucose tolerance. Post-challenge hyperinsulinaemia was associated with the number of diseased coronary arteries and the presence of previous MI. The insulin response to the glucose challenge test requires further investigation as a potential risk factor for CAD and a potential target for intervention. PMID- 15894764 TI - Massive air embolism after Maze. PMID- 15894765 TI - Long term outcome in patients with silent versus symptomatic ischaemia during dobutamine stress echocardiography. AB - OBJECTIVES: To compare the long term prognosis of patients having silent versus symptomatic ischaemia during dobutamine stress echocardiography (DSE). DESIGN: Observational study. SETTING: Tertiary referral centre. PATIENTS: 931 patients who experienced stress induced myocardial ischaemia during DSE. RESULTS: Silent ischaemia was present in 643 of 931 patients (69%). The number of dysfunctional segments at rest (mean (SD) 9.6 (5.1) v 8.8 (5.0), p = 0.1) and of ischaemic segments (3.5 (2.2) v 3.8 (2.1), p = 0.2) was comparable in both groups. During a mean (SD) follow up of 5.5 (3.3) years, there were 169 (18%) cardiac deaths and 86 (9%) non-fatal infarctions. Multivariable Cox regression analysis showed age (hazard ratio (HR) 1.1, 95% confidence interval (CI) 1.02 to 1.05), previous myocardial infarction (HR 1.4, 95% CI 1.1 to 2.0), and number of ischaemic segments during the test (HR 2.0, 95% CI 1.0 to 3.7) as independent predictors of cardiac death and myocardial infarction. For every additional ischaemic segment there was a twofold increment in risk of late cardiac events. The annual cardiac death or myocardial infarction rate was 3.0% in patients with symptomatic ischaemia and 4.6% in patients with silent ischaemia (p < 0.01). Silent induced ischaemia was an independent predictor of cardiac death and myocardial infarction (HR 1.7, 95% CI 1.1 to 2.0). During follow up symptomatic patients were treated more often with cardioprotective therapy (p < 0.01) and coronary revascularisation (145 of 288 (50%) v 174 of 643 (27%), p < 0.001). CONCLUSIONS: Patients with silent ischaemia had a similar extent of myocardial ischaemia during DSE compared to patients with symptomatic ischaemia but received less cardioprotective treatment and coronary revascularisation and experienced a higher cardiac event rate. PMID- 15894766 TI - Long term clinical and echocardiographic results of mitral balloon valvotomy in children and adolescents. AB - AIMS: To assess the safety, efficacy, and long term results of mitral balloon valvotomy (MBV) for rheumatic mitral stenosis in children and adolescents in comparison to adults. METHODS: The results of 468 patients with mitral stenosis who underwent successful MBV and were followed up for 0.5-13 years were analysed. Patients were divided according to age at the time of MBV into group 1 consisting of 84 patients < or = 20 years of age (children and adolescents) and group 2 that included 384 patients, age > 20 (adults). RESULTS: Patients in group 1 had a lower mitral echo score (mean (SD) 7.5 (1.3) v 8 (1.1), p < 0.001), smaller Doppler mitral valve area (MVA) (0.84 (0.17) v 0.92 (0.18) cm2, p < 0.001), and higher Doppler mitral valve gradient (15.0 (5.3) v 12.7 (4.5) mm Hg, p < 0.001) than group 2. Immediately after MBV group 1 had larger MVA, whether measured by Doppler (2.0 (0.30) v 1.96 (0.28) cm2, p < 0.05) or by catheter (2.0 (0.59) v 1.8 (0.52) cm2, p < 0.001), and similar complication rates, compared to group 2. After a mean follow up of 5 (3.5) years there was no significant difference between groups 1 and 2 in the incidence of restenosis (14.3% v 16.1%, NS). Event free survival rates at 5, 10, and 12.5 years were 93%, 79%, and 79% for group 1 and 94%, 90%, and 84% for group 2 (p = 0.18). CONCLUSIONS: MBV is safe and effective in children and adolescents with rheumatic mitral stenosis. It provides better immediate results than in adults and excellent long term results that are comparable to those seen in adults. PMID- 15894767 TI - Right ventricular outflow tract obstruction in an infant. PMID- 15894768 TI - Allopurinol reduces B-type natriuretic peptide concentrations and haemoglobin but does not alter exercise capacity in chronic heart failure. AB - OBJECTIVE: To study whether the effect of allopurinol on improvement of endothelial dysfunction in chronic heart failure (CHF) translates into improved exercise capacity and to examine whether allopurinol also improves B-type natriuretic peptide (BNP), the other important prognostic marker of CHF. DESIGN: Randomised, double blind, placebo controlled crossover trial. SETTING: Teaching hospital. PATIENTS: 50 patients with CHF (New York Heart Association functional classes II and III) were recruited. INTERVENTIONS: 50 patients with CHF were randomly assigned to three months' treatment with allopurinol (300 mg/day) or placebo. At two and three months into treatment, they underwent a modified Bruce exercise protocol and a six minute walk test. Blood was taken for BNP and haemoglobin analysis. RESULTS: Neither exercise test was altered by allopurinol. However, plasma BNP concentrations fell significantly (p = 0.035) with allopurinol (11.9 pmol/l) versus placebo (14.4 pmol/l). Haemoglobin concentrations also fell highly significantly with allopurinol (p = 0.001). CONCLUSIONS: An important negative finding is that despite high hopes for it, allopurinol had no effect on exercise capacity in CHF. On the other hand, allopurinol did reduce BNP, which is the best available surrogate marker for prognosis in CHF. PMID- 15894769 TI - Increased risk of heart failure as a consequence of perioperative myocardial injury after coronary artery bypass grafting. AB - OBJECTIVE: To analyse the relation between perioperative myocardial injury (PMI) and the risk of subsequent heart failure after coronary artery bypass grafting (CABG). DESIGN AND SETTING: Clinical data were documented prospectively in all patients and stored in a computer. All hospital readmissions were identified and the registered primary diagnoses were analysed. Survival information on all patients was obtained by use of combined registers. The study was carried out at the cardiac surgical referral centre of University Hospital, Uppsala, Sweden. PATIENTS: 7493 patients discharged alive after primary CABG between 1987 and 1996 were followed up until the first hospital readmission for heart failure, death, or 31 December 1996 was reached. MAIN OUTCOME MEASURES: Hospital readmission for heart failure or late mortality. RESULTS: Of the patients studied 576 (7.7%) were readmitted for heart failure. Actuarial freedom from readmission for heart failure after four years was 93%, and after seven years, 89%. Of the 576 patients, 114 (20%) had had PMI, which increased the risk of heart failure independently (hazard ratio (HR) 2.3, 95% confidence interval (CI) 1.8 to 2.8). Increased age, female sex, diabetes, previous myocardial infarction, dyspnoea, preoperative atrial fibrillation, left ventricular dysfunction, and triple vessel disease were independent risk factors for heart failure. The use of an internal mammary artery decreased the risk. PMI implied increased mortality (HR 1.4, 95% CI 1.1 to 1.8). Late mortality was greatly increased in patients readmitted for heart failure. CONCLUSION: PMI increased the risk of heart failure and late death after CABG, and heart failure had a notable adverse effect on late survival. PMID- 15894770 TI - Effects of congestive heart failure on plasma von Willebrand factor and soluble P selectin concentrations in patients with non-valvar atrial fibrillation. AB - OBJECTIVE: To examine further the relations of plasma von Willebrand factor (vWf, an index of endothelial damage and dysfunction) and soluble P-selectin (sP-sel, an index of platelet activation) concentrations to the presence and onset of clinical congestive heart failure (CHF) and the degree of left ventricular (LV) dysfunction in patients taking part in the SPAF (stroke prevention in atrial fibrillation) study. METHODS: Plasma concentrations of vWf and sP-sel were measured by enzyme linked immunosorbent assay (ELISA) in 1321 participants in the SPAF III study and related to the presence and onset of clinical CHF, as well as echocardiographic findings. Of the 1321 patients with atrial fibrillation (AF), 331 (25%) had a documented history of clinical heart failure, of which 168 cases were related to a new or recurrent episode of acute decompensated heart failure occurring within the preceding three months. RESULTS: Mean plasma vWf was higher among patients with AF and CHF (154 (29) v 144 (31) IU/dl, p < 0.001), particularly those with acute or recent decompensated symptoms. Patients with severe LV dysfunction on two dimensional echocardiography and low fractional shortening also had significantly higher vWf concentrations than those with no LV dysfunction. CHF patients with clinical features--with (156 (28) IU/dl) and without (152 (31) IU/dl) LV dysfunction--also had higher mean vWf concentrations than patients with asymptomatic LV dysfunction (146 (31) IU/dl, p < 0.001). The presence of mitral regurgitation in CHF was associated with lower vWf concentrations. Plasma sP-sel concentrations were not affected by presence, onset, or severity of heart failure. CONCLUSIONS: CHF may contribute to hypercoagulability and thrombotic risk in AF through increased endothelial damage and dysfunction. Patients with acute or recent decompensated features have the highest degree of endothelial damage and dysfunction. The presence of CHF clinical features was an important determinant of plasma vWf concentrations. PMID- 15894771 TI - Effects of ventricular rate and regularity on the velocity and magnitude of left atrial appendage flow in atrial fibrillation. AB - OBJECTIVE: To prospectively determine whether ventricular rate and regularity are significant determinants of the velocity and magnitude of left atrial appendage (LAA) flow. DESIGN AND PATIENTS: 12 patients with atrial fibrillation (AF), high degree atrioventricular block, and indwelling permanent pacemakers were studied. SETTING: Cardiology department of a tertiary referral centre. INTERVENTIONS: Pacing was triggered by an external programmable transcutaneous device. Patients were paced at 60, 120, and 150 beats/min in both regular and irregular rhythm. LAA flow velocity and magnitude were assessed with transoesophageal Doppler echocardiography. MAIN OUTCOME MEASURES: Peak and mean LAA inflow and outflow velocity, and time-velocity interval (TVI) of LAA flow. RESULTS: Increasing ventricular rate was associated with significantly lower peak inflow (p < 0.01), peak outflow (p < 0.05), mean inflow (p < 0.01), and mean outflow (p < 0.05) velocities and with a lower TVI of LAA filling and emptying velocities (p < 0.01). This effect was noted at rates of 60 beats/min compared with both 120 and 150 beats/min. At a pacing rate of 120 beats/min there was a significantly higher total TVI when pacing at a regular than at an irregular rhythm (40.16 (14.6) cm v 30.74 (10.9) cm, p < 0.05). CONCLUSIONS: In this study, LAA filling velocities in patients in AF were significantly influenced by paced ventricular rate and to a much lesser extent ventricular rhythm. These results suggest that rapid ventricular rates may predispose to stasis in the LAA in AF. PMID- 15894773 TI - A vascular ring without compression: double aortic arch presenting as a coincidental finding during cardiac catheterisation. PMID- 15894772 TI - Ventriculo-vascular interactions in patients with beta thalassaemia major. AB - OBJECTIVES: To determine potential interactions between the heart and arterial system in patients with beta thalassaemia major. DESIGN AND PATIENTS: Vascular compliance, systemic vascular resistance, and left ventricular (LV) contractility was determined in 34 asymptomatic thalassaemia patients at 2-4 hours after blood transfusion and also in 34 age and sex matched controls using a non-invasive device. The results were compared between groups and inter-relationships between LV contractility and indices of vascular load were explored. SETTING: Tertiary paediatric cardiac centre. RESULTS: When compared with controls, patients had greater systemic vascular resistance (1633 (259) v 1377 (276) dynes/s/cm5, p < 0.001) and effective arterial elastance (E(a)) (1.86 (0.25) v 1.65 (0.29) mm Hg/ml, p = 0.001), an index of combined pulsatile and static vascular load. On the other hand, their systolic blood pressure (104 (9) v 112 (13) mm Hg, p = 0.006), pulse pressure (45 (9) v 57 (10) mm Hg, p < 0.001), adjusted systemic vascular compliance (1.21 (0.09) v 1.37 (0.14), p < 0.001), adjusted brachial artery distensibility (21 (0.29) v 7.95 (0.29)%/mm Hg, p < 0.001) and LV+dP/dt (1059 (183) v 1239 (237) mm Hg/s, p = 0.001) were significantly lower. Significant determinants of LV contractility, as reflected by LV+dP/dt, were age (standardised beta = -0.24, p = 0.003), body mass index (standardised beta = 0.34, p = 0.004), systolic blood pressure (standardised beta = 0.90, p < 0.001), and effective E(a) (standardised beta = -0.50, p < 0.001) (model R2 = 0.69). No significant correlation existed between serum ferritin concentration and any of the cardiac or vascular indices. CONCLUSION: An unfavourable ventriculo-vascular interaction, as characterised by impaired cardiac contractility and increased static and pulsatile vascular load, occurs in patients with beta thalassaemia major. PMID- 15894774 TI - Left ventricular pseudoaneurysm in a child. PMID- 15894775 TI - N-acetylcysteine does not prevent contrast induced nephropathy after cardiac catheterisation with an ionic low osmolality contrast medium: a multicentre clinical trial. AB - OBJECTIVE: To evaluate oral N-acetylcysteine in the prevention of contrast induced nephropathy (CIN) in patients at low to moderate risk undergoing cardiac catheterisation with ionic low osmolality contrast medium. METHODS: In a multicentre double blind clinical trial 156 patients undergoing coronary angiography or percutaneous coronary intervention with serum creatinine > or = 106.08 micromol/l or creatinine clearance < 50 ml/min or diabetes mellitus were randomly assigned to receive N-acetylcysteine 600 mg orally twice daily for two days or placebo. Only low osmolality ionic contrast medium was used. RESULTS: Sixteen patients developed CIN, defined as an increase of 44.2 micromol/l in creatinine in 48 hours: eight of 77 patients (10.4%) in the N-acetylcysteine group and eight of 79 patients (10.1%) in the placebo group (p = 1.00). The mean (SD) change in serum creatinine was similar in both groups: 7.96 (35.36) micromol/l in the N-acetylcysteine group and 6.19 (25.64) micromol/l in the placebo group (p = 0.67). No difference was observed in the change in endogenous creatinine clearance (-0.54 (10.4) ml/min v -2.52 (12.3) ml/min, N-acetylcysteine and placebo, respectively, p = 0.28). CONCLUSION: Oral N-acetylcysteine did not prevent CIN in patients at low to moderate risk undergoing cardiac catheterisation with ionic low osmolality contrast medium. PMID- 15894776 TI - Transoesophageal echocardiography for the identification of a giant aortic thrombus. PMID- 15894777 TI - Predictive value of EuroSCORE on long term outcome in cardiac surgery patients: a single institution study. AB - OBJECTIVES: To assess the value of the European system for cardiac operative risk evaluation (EuroSCORE), a validated model for prediction of in-hospital mortality after cardiac surgery, in predicting long term event-free survival. DESIGN AND SETTING: Single institution observational cohort study. PATIENTS: Adult patients (n = 1230) who underwent cardiac surgery between January 2000 and August 2002. RESULTS: Mean age was 65 (11) years and 32% were women. Type of surgery was isolated coronary artery bypass grafting in 62%, valve surgery in 23%, surgery on the thoracic aorta in 4%, and combined or other procedures in 11%. Mean EuroSCORE was 4.53 (3.16) (range 0-21); 366 were in the low (0-2), 442 in the medium (3-5), 288 in the high (6-8), and 134 in the very high risk group (> or = 9). Information on deaths or events leading to hospital admission after the index discharge was obtained from the Regional Health Database. Out of hospital deaths were identified through the National Death Index. In-hospital 30 day mortality was 2.8% (n = 34). During 2024 person-years of follow up, 44 of 1196 patients discharged alive (3.7%) died. By Cox multivariate analysis, EuroSCORE was the single best independent predictor of long term all cause mortality (hazard ratio (HR) 1.55, 95% confidence interval (CI) 1.03 to 2.34, p < 0.0001). In the time to first event analysis, 227 either died without previous events (n = 20, 9%) or were admitted to hospital for an event (n = 207, 91%). EuroSCORE (HR 1.60, 95% CI 1.36 to 1.89, p < 0.0001), the presence of > or = 2 co-morbidities versus one (HR 1.49, 95% CI 1.09 to 2.02, p < 0.0001), and > 96 hours' stay in the intensive care unit after surgery (HR 2.04, 95% CI 1.42 to 2.95, p = 0.0001) were independently associated with the combined end point of death or hospital admission after the index discharge. CONCLUSIONS: EuroSCORE and a prolonged intensive care stay after surgery are associated with long term event-free survival and can be used to tailor long term postoperative follow up and plan resource allocation for the cardiac surgical patient. PMID- 15894778 TI - Arrhythmogenic right ventricular cardiomyopathy with fibrofatty atrophy, myocardial oedema, and aneurysmal dilation. PMID- 15894779 TI - Subxiphoid pericardiostomy in the management of pericardial effusions: case series analysis of 368 patients. AB - OBJECTIVE: To assess the effectiveness of subxiphoid pericardiostomy in the treatment and diagnosis of pericardial effusions. METHODS: 368 patients who underwent subxiphoid pericardiostomy and tube drainage for cardiac tamponade, moderate to severe pericardial effusion, or suspicious bacterial aetiology were retrospectively analysed. Biopsies of the pericardium and fluid samples for diagnostic tests were obtained from each patient. RESULTS: The mean age of the patients was 38.4 years, and the male to female ratio was 220:148. The pericardial effusion was classified by echocardiography as severe in 53% of the patients, moderate in 43%, and mild in 4%. The incidence of cardiac tamponade was 25%. Myocardial injury requiring sternotomy occurred as an operative complication in 0.8% of the patients and recurrent effusion necessitating further surgical intervention developed in 10% of patients. Histopathological examination and the polymerase chain reaction of specimens of pericardium and fluid were helpful for establishing a diagnosis in 90% of patients with malignancy and 92% of patients with tuberculous pericarditis. The overall 30 day mortality rate was 0.8%. Patients were followed up for at least one year. Pericardial constriction requiring pericardiectomy developed in 3% of the patients. CONCLUSIONS: Pericardial effusions of various causes can be safely, effectively, and quickly managed with subxiphoid pericardiostomy in both adults and children. PMID- 15894780 TI - Severe mitral regurgitation caused by annular abscess fistulating into the left atrium. PMID- 15894781 TI - Thoracoscopic ASD closure is a reliable supplement for percutaneous treatment. AB - OBJECTIVE: To determine the feasibility and effectiveness of endoscopic atrial septal defect (ASD) closure when percutaneous ASD closure is impossible or has failed. PATIENTS: Between March 1997 and February 2003, 74 patients (63% female, mean (SD) age 44 (16) years) underwent an endoscopic ASD closure. Median preoperative New York Heart Association functional class was I. Clinical and echocardiographic follow up was obtained for all patients (mean (SD) 38 (19) months). Patients were assessed for scar aesthetics, procedure related pain, functional recovery, and overall patient satisfaction. RESULTS: ASD closure was successful in all patients (two primum ASD, 68 secundum ASD, four sinus venosus type). Patch repair was performed in 42%. Mean aortic cross clamp and cardiopulmonary bypass times were 54 (24) minutes and 98 (35) minutes, respectively. There were no in-hospital deaths and no conversions to sternotomy. Complications included one iliac vein stenting, one femoral arterioplasty, two revisions for suspected bleeding, and seven cases of atrial fibrillation. Two patients required late reoperation: one for atrial thrombus and another for tricuspid regurgitation. Echocardiographic control confirmed complete ASD closure in 71 patients and a small residual shunt in three patients. Ninety three per cent of the patients were highly satisfied with very low procedure related pain and 97% felt they had an aesthetically pleasing scar. CONCLUSION: Endoscopic ASD closure can be safely done with a high degree of patient satisfaction. It is now the authors' exclusive surgical approach whenever percutaneous treatment is not indicated or has failed. PMID- 15894782 TI - Infective endocarditis in congenital heart disease: Japanese national collaboration study. AB - OBJECTIVE: To provide pure cohorts of paediatric and adult patients with congenital heart disease (CHD) and infective endocarditis (IE) for making future guidelines. DESIGN: Japanese nationwide survey. SETTING: 66 Japanese institutions. PATIENTS: 170 children, mean (SD) age 7.4 (5.7) years (range 14 days to 17 years), and 69 adults, age 32.5 (14.1) years (range 18-69) who developed IE between 1997 and 2001 (one in 240 admissions with CHD). MAIN OUTCOME MEASURES: Clinical presentation of IE. RESULTS: 119 patients including 88 with cyanotic CHD had previous cardiac surgery. Procedures preceding IE were dental (12%) followed by cardiovascular surgery (8%). Sites of infection were left sided in 46% and right sided in 51%. Vegetation with diameter of 11 mm was documented in 151 (63%). Frequent complications were embolic events (stroke 11%, other emboli 20%) and cardiac failure (23%). The most common microorganisms were streptococci (50%) and staphylococci (37%) with methicillin resistant Staphylococcus aureus in 7.5%. Empirical treatments were penicillins (alone or with other antibiotics 57%) followed by cephems (22%) and vancomycin (11%). Surgery during active IE was common (26%), with vegetation (45%) and heart failure (29%) as the most frequent indications. Mortality was 8.8%: 8.0% among patients who received medical treatment alone and 11.1% among those with active IE who underwent surgery. The causes of death (n = 21) were surgery (7), infection (7), cardiac failure (6), and renal failure (1). CONCLUSIONS: Because of a recent increase in the incidence of IE and high mortality and complication rate, it is mandatory to establish well formulated recommendations for management of IE in paediatric and adult patients with CHD based on a large cohort. Results of this nationwide multicentre database should be helpful in establishing guidelines. PMID- 15894783 TI - Pregnancy, fertility, and recurrence risk in corrected tetralogy of Fallot. AB - OBJECTIVE: To determine in women with surgically corrected tetralogy of Fallot the risk of pregnancy for mother and fetus, whether fertility was compromised, and the recurrence risk of congenital heart disease. DESIGN: Data were collected from 83 patients through interviews and review of medical records. RESULTS: In 29 patients 63 pregnancies were observed, of which 13 ended in an abortion. Fifty successful pregnancies were observed in 26 patients. During six successful pregnancies (12%) complications (symptomatic right sided heart failure, arrhythmias, or both) occurred. Both patients who developed symptomatic heart failure had severe pulmonary regurgitation. No clear relation between offspring mortality, premature birth or being small for gestational age, and cardiac characteristics of the mother was identified. Fifty seven patients were childless (41 (72%) voluntarily). Recurrence risk for congenital heart disease was 2.2%. Infertility was uncommon. CONCLUSIONS: Although complications did occur in five of 26 (19%) of the patients with a corrected tetralogy of Fallot, pregnancy was generally well tolerated in this largest report so far. No obvious predictors for maternal events or child outcome were determined, except for a possible relation between severe pulmonary regurgitation and symptomatic heart failure. PMID- 15894784 TI - Chest x rays illustrating successful resynchronisation therapy. PMID- 15894785 TI - Atorvastatin inhibits calcification and enhances nitric oxide synthase production in the hypercholesterolaemic aortic valve. AB - OBJECTIVE: To study in a rabbit model the expression of endothelial nitric oxide synthase (eNOS) in association with the development of calcification of the aortic valve, and to assess the effects of atorvastatin on eNOS expression, nitrite concentration, and aortic valve calcification. METHODS: Rabbits (n = 48) were treated for three months: 16, forming a control group, were fed a normal diet; 16 were fed a 0.5% (wt/wt) high cholesterol diet; and 16 were fed a 0.5% (wt/wt) cholesterol diet plus atorvastatin (2.5 mg/kg/day). The aortic valves were examined with eNOS immunostains and western blotting. Cholesterol and high sensitivity C reactive protein (hsCRP) concentrations were determined by standard assays. Serum nitrite concentrations were measured with a nitric oxide analyser. eNOS was localised by electron microscopy and immunogold labelling. Calcification in the aortic valve was evaluated by micro-computed tomography (CT). RESULTS: Cholesterol, hsCRP, and aortic valve calcification were increased in the cholesterol fed compared with control animals. Atorvastatin inhibited calcification in the aortic valve as assessed by micro-CT. eNOS protein concentrations were unchanged in the control and cholesterol groups but increased in the atorvastatin treated group. Serum nitrite concentrations were decreased in the hypercholesterolaemic animals and increased in the group treated with atorvastatin. CONCLUSION: These data provide evidence that chronic experimental hypercholesterolaemia produces bone mineralisation in the aortic valve, which is inhibited by atorvastatin. PMID- 15894786 TI - Left atrial membrane, left ventricular dysfunction, and mitral regurgitation in an adult. PMID- 15894787 TI - Quantitative assessment of regional peak myocardial acceleration during isovolumic contraction and relaxation times by tissue Doppler imaging. AB - OBJECTIVE: To examine regional wall acceleration and its relation to relaxation. STUDY DESIGN: 8 sheep were examined by tissue Doppler ultrasound imaging (VingMed Vivid FiVe) in apical four chamber views to evaluate the left ventricular wall divided into six segments and the mitral annulus in two segments. Peak myocardial acceleration during isovolumic periods (pIVA) derived from tissue Doppler echocardiography was analysed during isovolumic contraction (ICT) and relaxation times (IRT) in each segment. INTERVENTIONS: After scanning at baseline, haemodynamic status was changed by administration of blood, dobutamine, and metoprolol. Changes of pIVA during IRT and ICT were compared over the four haemodynamic conditions in parallel with their peak positive and negative dP/dt measured with a high frequency manometer tipped catheter. RESULTS: pIVA of the basal lateral segment during ICT correlated most strongly with peak positive dP/dt (r = 0.96, p < 0.0001) and there was good correlation between pIVA of the mitral valve annulus in the septum during IRT and peak negative dP/dt (r = 0.80, p < 0.0001). pIVA differed significantly between the four haemodynamic conditions during ICT in all segments (p < 0.05); pIVA during IRT did not differ significantly between the four conditions. CONCLUSIONS: pIVA of the basal lateral wall during ICT correlated most strongly with peak positive dP/dt, and pIVA of the septal mitral valve annulus during IRT correlated well with peak negative dP/dt. PMID- 15894788 TI - Impact of asynchronous ventricular activation on pro-inflammatory cytokines and oxidative stress in paced patients. PMID- 15894789 TI - Prognostic implications of sonographic characteristics of carotid plaques in patients with acute coronary syndromes. PMID- 15894790 TI - Fibrinolytic treatment for recurrent left sided prosthetic valve thrombosis. PMID- 15894791 TI - Repeated beta irradiation for failed intracoronary radiation therapy in patients with in-stent restenosis. PMID- 15894792 TI - Endothelin antagonism in pulmonary hypertension, heart failure, and beyond. PMID- 15894793 TI - Inter-professional team approach to patients with heart failure. PMID- 15894794 TI - The Fontan circulation. PMID- 15894795 TI - Identification of CELF splicing activation and repression domains in vivo. AB - CUG-BP and ETR-3 like factor (CELF) proteins are regulators of pre-mRNA alternative splicing. We created a series of truncation mutants to identify the regions of CELF proteins that are required to activate and to repress alternative splicing of different exons. This analysis was performed in parallel on two CELF proteins, ETR-3 (CUG-BP2, NAPOR, BRUNOL3) and CELF4 (BRUNOL4). We identified a 20 residue region of CELF4 required for repression or activation, in contrast to ETR 3, for which the required residues are more disperse. For both ETR-3 and CELF4, distinct regions were required to activate splicing of two different alternative exons, while regions required for repression of an additional third exon overlapped with regions required for activation. Our results suggest that activation of different splicing events by individual CELF proteins requires separable regions, implying the nature of the protein-protein interactions required for activation are target-dependent. The finding that residues required for activation and repression overlap suggests either that the same region interacts with different proteins to mediate different effects or that interactions with the same proteins can have different effects on splicing due to yet-to-be defined downstream events. These results provide a foundation for identifying CELF-interacting proteins involved in activated and/or repressed splicing. PMID- 15894796 TI - Unusual features of fibrillarin cDNA and gene structure in Euglena gracilis: evolutionary conservation of core proteins and structural predictions for methylation-guide box C/D snoRNPs throughout the domain Eucarya. AB - Box C/D ribonucleoprotein (RNP) particles mediate O2'-methylation of rRNA and other cellular RNA species. In higher eukaryotic taxa, these RNPs are more complex than their archaeal counterparts, containing four core protein components (Snu13p, Nop56p, Nop58p and fibrillarin) compared with three in Archaea. This increase in complexity raises questions about the evolutionary emergence of the eukaryote-specific proteins and structural conservation in these RNPs throughout the eukaryotic domain. In protists, the primarily unicellular organisms comprising the bulk of eukaryotic diversity, the protein composition of box C/D RNPs has not yet been extensively explored. This study describes the complete gene, cDNA and protein sequences of the fibrillarin homolog from the protozoon Euglena gracilis, the first such information to be obtained for a nucleolus localized protein in this organism. The E.gracilis fibrillarin gene contains a mixture of intron types exhibiting markedly different sizes. In contrast to most other E.gracilis mRNAs characterized to date, the fibrillarin mRNA lacks a spliced leader (SL) sequence. The predicted fibrillarin protein sequence itself is unusual in that it contains a glycine-lysine (GK)-rich domain at its N terminus rather than the glycine-arginine-rich (GAR) domain found in most other eukaryotic fibrillarins. In an evolutionarily diverse collection of protists that includes E.gracilis, we have also identified putative homologs of the other core protein components of box C/D RNPs, thereby providing evidence that the protein composition seen in the higher eukaryotic complexes was established very early in eukaryotic cell evolution. PMID- 15894797 TI - Automated band mapping in electrophoretic gel images using background information. AB - Some popular methods for polymorphism and mutation discovery involve ascertainment of novel bands by the examination of electrophoretic gel images. Although existing strategies for mapping bands work well for specific applications, such as DNA sequencing, these strategies are not well suited for novel band detection. Here, we describe a general strategy for band mapping that uses background banding patterns to facilitate lane calling and size calibration. We have implemented this strategy in GelBuddy, a user-friendly Java-based program for PC and Macintosh computers, which includes several utilities to assist discovery of mutations and polymorphisms. We demonstrate the use of GelBuddy in applications based on single-base mismatch cleavage of heteroduplexed PCR products. Use of software designed to facilitate novel band detection can significantly shorten the time needed for image analysis and data entry in a high throughput setting. Furthermore, the interactive strategy implemented in GelBuddy has been successfully applied to DNA fingerprinting applications, such as AFLP. GelBuddy promises to make electrophoretic gel analysis a viable alternative to DNA resequencing for discovery of mutations and polymorphisms. PMID- 15894798 TI - Ferredoxin-NADP+ reductase. Kinetics of electron transfer, transient intermediates, and catalytic activities studied by flash-absorption spectroscopy with isolated photosystem I and ferredoxin. AB - The electron transfer cascade from photosystem I to NADP+ was studied at physiological pH by flash-absorption spectroscopy in a Synechocystis PCC6803 reconstituted system comprised of purified photosystem I, ferredoxin, and ferredoxin-NADP+ reductase. Experiments were conducted with a 34-kDa ferredoxin NADP+ reductase homologous to the chloroplast enzyme and a 38-kDa N-terminal extended form. Small differences in kinetic and catalytic properties were found for these two forms, although the largest one has a 3-fold decreased affinity for ferredoxin. The dissociation rate of reduced ferredoxin from photosystem I (800 s(-1)) and the redox potential of the first reduction of ferredoxin-NADP+ reductase (-380 mV) were determined. In the absence of NADP+, differential absorption spectra support the existence of a high affinity complex between oxidized ferredoxin and semireduced ferredoxin-NADP+ reductase. An effective rate of 140-170 s(-1) was also measured for the second reduction of ferredoxin-NADP+ reductase, this process having a rate constant similar to that of the first reduction. In the presence of NADP+, the second-order rate constant for the first reduction of ferredoxin-NADP+ reductase was 20% slower than in its absence, in line with the existence of ternary complexes (ferredoxin-NADP+ reductase)-NADP+ ferredoxin. A single catalytic turnover was monitored, with 50% NADP+ being reduced in 8-10 ms using 1.6 microM photosystem I. In conditions of multiple turnover, we determined initial rates of 360-410 electrons per s and per ferredox in-NADP+ reductase for the reoxidation of 3.5 microM photoreduced ferredoxin. Identical rates were found with photosystem I lacking the PsaE subunit and wild type photosystem I. This suggests that, in contrast with previous proposals, the PsaE subunit is not involved in NADP+ photoreduction. PMID- 15894799 TI - Regiochemistry of neuroprostanes generated from the peroxidation of docosahexaenoic acid in vitro and in vivo. AB - Isoprostanes (IsoPs) are isomers of prostaglandins that are generated from the free radical-initiated peroxidation of arachidonic acid (C20.4 omega-6). IsoPs exert potent bioactivity and are regarded as the "gold standard" to assess oxidative stress in various human diseases. Analogously, autoxidation of docosahexaenoic acid (DHA, C22.6 omega-3) generates an array of IsoP-like compounds that are termed neuroprostanes (NPs). A major class of NPs identified in vitro and in vivo contains F-type prostane rings and are know as F4-NPs. A number of different F4-NP regioisomers are formed from the peroxidation of DHA. Among the eight possible regioisomeric groups, we hypothesize that 4- and 20 series NPs are generated in greater amounts than other classes because the precursors that lead to regioisomers other than those of the 4- and 20-series can be further oxidized to form novel dioxolane-IsoP-like compounds, analogous to those generated from arachidonate. Various mass spectrometric approaches, including electron capture atmospheric pressure chemical ionization mass spectrometry, were utilized to analyze NPs formed in vitro and in vivo based on their characteristic fragmentation in the gas phase. Experimental results were consistent with our hypothesis that 4- and 20-series NP regioisomers are preferentially generated. The discovery of regioselectivity in the formation of NPs will allow studies of the biological activities of NPs to focus on the more abundantly generated compounds to determine their role in modulating the pathophysiological consequences of DHA oxidation and oxidant stress. PMID- 15894800 TI - Thiocyanate modulates the catalytic activity of mammalian peroxidases. AB - We investigated the potential role of the co-substrate, thiocyanate (SCN-), in modulating the catalytic activity of myeloperoxidase (MPO) and other members of the mammalian peroxidase superfamily (lactoperoxidase (LPO) and eosinophil peroxidase (EPO)). Pre-incubation of SCN- with MPO generates a more complex biological setting, because SCN- serves as either a substrate or inhibitor, causing diverse impacts on the MPO heme iron microenvironment. Consistent with this hypothesis, the relationship between the association rate constant of nitric oxide binding to MPO-Fe(III) as a function of SCN- concentration is bell-shaped, with a trough comparable with normal SCN- plasma levels. Rapid kinetic measurements indicate that MPO, EPO, and LPO Compound I formation occur at rates slower than complex decay, and its formation serves to simultaneously catalyze SCN- via 1e- and 2e- oxidation pathways. For the three enzymes, Compound II formation is a fundamental feature of catalysis and allows the enzymes to operate at a fraction of their possible maximum activities. MPO and EPO Compound II is relatively stable and decays gradually within minutes to ground state upon H2O2 exhaustion. In contrast, LPO Compound II is unstable and decays within seconds to ground state, suggesting that SCN- may serve as a substrate for Compound II. Compound II formation can be partially or completely prevented by increasing SCN- concentration, depending on the experimental conditions. Collectively, these results illustrate for the first time the potential mechanistic differences of these three enzymes. A modified kinetic model, which incorporates our current findings with the mammalian peroxidases classic cycle, is presented. PMID- 15894801 TI - Ryanodine receptors in muscarinic receptor-mediated bronchoconstriction. AB - Ryanodine receptors (RyRs), intracellular calcium release channels essential for skeletal and cardiac muscle contraction, are also expressed in various types of smooth muscle cells. In particular, recent studies have suggested that in airway smooth muscle cells (ASMCs) provoked by spasmogens, stored calcium release by the cardiac isoform of RyR (RyR2) contributes to the calcium response that leads to airway constriction (bronchoconstriction). Here we report that mouse ASMCs also express the skeletal muscle and brain isoforms of RyRs (RyR1 and RyR3, respectively). In these cells, RyR1 is localized to the periphery near the cell membrane, whereas RyR3 is more centrally localized. Moreover, RyR1 and/or RyR3 in mouse airway smooth muscle also appear to mediate bronchoconstriction caused by the muscarinic receptor agonist carbachol. Inhibiting all RyR isoforms with > or = 200 microM ryanodine attenuated the graded carbachol-induced contractile responses of mouse bronchial rings and calcium responses of ASMCs throughout the range of carbachol used (50 nM to > or = 3 microM). In contrast, inhibiting only RyR1 and RyR3 with 25 microM dantrolene attenuated these responses caused by high (>500 nM) but not by low concentrations of carbachol. These data suggest that, as the stimulation of muscarinic receptor in the airway smooth muscle increases, RyR1 and/or RyR3 also mediate the calcium response and thus bronchoconstriction. Our findings provide new insights into the complex calcium signaling in ASMCs and suggest that RyRs are potential therapeutic targets in bronchospastic disorders such as asthma. PMID- 15894802 TI - Protein kinase C phosphorylation of the metabotropic glutamate receptor mGluR5 on Serine 839 regulates Ca2+ oscillations. AB - The activation of Group 1 metabotropic glutamate receptors, mGluR5 and mGluR1alpha, triggers intracellular calcium release; however, mGluR5 activation is unique in that it elicits Ca2+ oscillations. A short region of the mGluR5 C terminus is the critical determinant and differs from the analogous region of mGluR1alpha by a single amino acid residue, Thr-840, which is an aspartic acid (Asp-854) in mGluR1alpha. Previous studies show that mGluR5-elicited Ca2+ oscillations require protein kinase C (PKC)-dependent phosphorylation and identify Thr-840 as the phosphorylation site. However, direct phosphorylation of mGluR5 has not been studied in detail. We have used biochemical analyses to directly investigate the phosphorylation of the mGluR5 C terminus. We showed that Ser-839 on mGluR5 is directly phosphorylated by PKC, whereas Thr-840 plays a permissive role. Although Ser-839 is conserved in mGluR1alpha (Ser-853), it is not phosphorylated, as the adjacent residue (Asp-854) is not permissive; however, mutagenesis of Asp-854 to a permissive alanine residue allows phosphorylation of Ser-853 on mGluR1alpha. We investigated the physiological consequences of mGluR5 Ser-839 phosphorylation using Ca2+ imaging. Mutations that eliminate Ser-839 phosphorylation prevent the characteristic mGluR5-dependent Ca2+ oscillations. However, mutation of Thr-840 to alanine, which prevents potential Thr-840 phosphorylation but is still permissive for Ser-839 phosphorylation, has no effect on Ca2+ oscillations. Thus, we showed that it is phosphorylation of Ser 839, not Thr-840, that is absolutely required for the unique Ca2+ oscillations produced by mGluR5 activation. The Thr-840 residue is important only in that it is permissive for the PKC-dependent phosphorylation of Ser-839. PMID- 15894803 TI - Nicotinic acetylcholine receptor subunits and associated proteins in human sperm. AB - We demonstrated previously the involvement of a nicotinic acetylcholine receptor containing an alpha7 subunit in the human sperm acrosome reaction (a modified exocytotic event essential to fertilization). Here we report the presence in human sperm of alpha7, alpha9, alpha3, alpha5, and beta4 nicotinic acetylcholine receptor subunits and the following proteins known to be associated with the receptor in the somatic cell: rapsyn and the tyrosine kinases c-SRC and FYN. The alpha7 subunit appears to exist as a homomer in the posterior post-acrosomal and neck regions of sperm and is probably linked to the cytoskeleton via rapsyn. The alpha3, alpha5, and beta4 subunits are present in the sperm flagellar mid-piece of sperm and possibly exist as alpha3alpha5beta4 and/or alpha3beta4 channels. The alpha9 subunit is present in the sperm mid-piece. We detected the FYN and c-SRC tyrosine kinases in the flagellar mid-piece region. Both co-precipitated only with the nicotinic acetylcholine receptor beta4 subunit. Immunolocalization with a C-terminal SRC kinase antibody, which recognizes several members of SRC kinase family, detected a SRC kinase co-localized with the alpha7 subunit in the neck region of sperm. Immunoprecipitation studies with that antibody demonstrated that the alpha7 subunit is associated with a SRC kinase. Antagonists of tyrosine phosphorylation inhibited the acetylcholine-initiated acrosome reaction, suggesting the involvement of a SRC kinase in the acrosome reaction. PMID- 15894804 TI - Translocation of dynorphin neuropeptides across the plasma membrane. A putative mechanism of signal transmission. AB - Several peptides, including penetratin and Tat, are known to translocate across the plasma membrane. Dynorphin opioid peptides are similar to cell-penetrating peptides in a high content of basic and hydrophobic amino acid residues. We demonstrate that dynorphin A and big dynorphin, consisting of dynorphins A and B, can penetrate into neurons and non-neuronal cells using confocal fluorescence microscopy/immunolabeling. The peptide distribution was characterized by cytoplasmic labeling with minimal signal in the cell nucleus and on the plasma membrane. Translocated peptides were associated with the endoplasmic reticulum but not with the Golgi apparatus or clathrin-coated endocytotic vesicles. Rapid entry of dynorphin A into the cytoplasm of live cells was revealed by fluorescence correlation spectroscopy. The translocation potential of dynorphin A was comparable with that of transportan-10, a prototypical cell-penetrating peptide. A central big dynorphin fragment, which retains all basic amino acids, and dynorphin B did not enter the cells. The latter two peptides interacted with negatively charged phospholipid vesicles similarly to big dynorphin and dynorphin A, suggesting that interactions of these peptides with phospholipids in the plasma membrane are not impaired. Translocation was not mediated via opioid receptors. The potential of dynorphins to penetrate into cells correlates with their ability to induce non-opioid effects in animals. Translocation across the plasma membrane may represent a previously unknown mechanism by which dynorphins can signal information to the cell interior. PMID- 15894805 TI - PDGF-BB regulates p27 expression through ERK-dependent RNA turn-over in vascular smooth muscle cells. AB - Cyclin-dependent kinase inhibitor p27, a critical determinant for cell cycle progression, is an important regulation target of mitogenic signals during arterial injury. In this study, we show in rat aortic smooth muscle cells that PDGF-BB down-regulated p27 protein and mRNA in an ERK-dependent mechanism. Inhibition of ERK, but not other subtypes of the mitogen-activated protein kinase family, prevented the reduction of p27 protein and mRNA. Conversely, direct activation of ERK via adenovirus-mediated expression of a constitutively active form of MEK led to a reduction of p27 protein and mRNA, further supporting the central role of ERK in regulation of p27 expression. Rapamycin, which potently inhibited PDGF-induced activation of p70 S6 kinase as well as proliferation of smooth muscle cells, did not alter the expression of p27. To delineate the molecular mechanism underlying the p27 down-regulation, we examined the effect of PDGF-BB on p27 promoter activity as well as mRNA stability. Stimulation with PDGF BB significantly shortened the half-life of p27 mRNA without affecting its promoter activity. To further understand the PDGF-stimulated p27 mRNA turnover, we inserted the 5'- and/or 3'-untranslated regions of p27 cDNA into a non-PDGF responsive luciferase gene. Only those chimeric genes that contained the 3' untranslated region responded to PDGF-BB with reduced expression. Moreover, inhibition of ERK completely prevented the effect of PDGF on the chimera expression. In summary, our data suggest that p27 is down-regulated by PDGF-BB in vascular smooth muscle cells through an ERK-dependent posttranscriptional mechanism. PMID- 15894806 TI - Expression of two genes encoding gibberellin 2- and 3-oxidases in developing seeds of Phaseolus coccineus. AB - We have isolated PcGA3ox1, a cDNA clone from developing runner bean (Phaseolus coccineus) seeds that shows significant amino acid homology with the gibberellin (GA) 3-oxidases. A recombinant fusion protein of PcGA3ox1 converted GA20 and GA9 to GA1 and GA4, respectively. In situ hybridization results showed that transcripts of this gene accumulate specifically within the suspensor of globular stage embryos. PcGA3ox1 mRNA begins to accumulate in the epidermal cells of the embryo proper and is also detectable in the endosperm during the transition from globular- to heart-stage embryos. PcGA3ox1 transcripts were localized exclusively in the cotyledons from the early cotyledonary stage up to the cotyledonary stage. Transcripts of the previously cloned GA 2-oxidase (PcGA2ox1) from developing seeds of runner bean were found primarily within the suspensor neck region from the late globular stage up to the heart stage. PcGA2ox1 mRNA was detectable in the whole suspensor from the early cotyledonary stage, and was found in the inner layer of integuments at the cotyledonary stage. Soluble enzyme preparations made from suspensors and embryos at two stages of embryogenesis (the heart and cotyledonary stages) were incubated with [14C]GA20 and [14C]GA1. Only young suspensor preparations converted GA20 to GA1 and GA5. Both suspensor preparations converted GA1 to GA8. Both embryo preparations converted GA20 to GA1, but were unable to convert GA1 to GA8. PMID- 15894807 TI - Living donor transplantation in the USA: are there any lessons for Europe? PMID- 15894808 TI - Angiotensin II down-regulates the SR-BI HDL receptor in proximal tubular cells. AB - BACKGROUND: The kidney plays an important role in the metabolism of lipoproteins, but renal cells are also a target of lipids under pathophysiological conditions contributing to organ damage and progression of disease. The majority of studies has focused on the interaction of renal cells with low-density lipoproteins. Relatively little is known of potential metabolism of high-density lipoproteins (HDL) on renal cells However, diverse pathophysiological situations, such as the nephrotic syndrome and acute renal injury, may be associated with an activated renin-angiotensin system as well as altered renal handling of HDL. Therefore, the present study sought to gain insight into the expression of the HDL receptor scavenger receptor class B type I (SR-BI) in cultured renal cells and a potential regulation by angiotensin II (ANG II). METHODS: Different renal cells lines and primary cultures (proximal tubular and mesangial cells) were screened by western blot for the expression of SR-BI. MCT cells, a mouse proximal tubular cell line, were selected for further studies. SR-BI protein and mRNA expression were determined after treatment with various doses of ANG II in the presence or absence of AT(1)- or AT(2)-receptor blocker. Uptake of HDL-associated cholesteryl ester into MCT cells was determined. Finally, rats were infused intraperitoneally with ANG II for 3-7 days, proximal tubules were isolated by differential centrifugation and SR-BI protein expression was assessed. Results. SR-BI protein was expressed in various primary cultures and permanent renal cell lines. ANG II (10(-10)-10(-6) M) treatment for 24 h induced a significant down-regulation of SR BI protein and mRNA expression in MCT cells. This suppression was attenuated by an AT(1)-receptor antagonist whereas an AT(2)-blocker was without effect. MCT cells revealed a high selective uptake of HDL cholesteryl ester that was significantly higher than that in syngeneic mesangial cells. ANG II for 24 h significantly reduced this selective HDL cholesteryl ester uptake into MCT, but not mesangial cells. Finally, ANG II- infusion into rats for 3 and 7 days induced a significant decrease of SR-BI protein expression in isolated tubules. CONCLUSIONS: Our data show that ANG II mediates down-regulation of SR-BI expression on proximal tubular cells in vivo and in vitro. However, the effects were small and additional experiments are necessary to confirm these first observations. The attenuated SR-BI expression is functionally relevant and associated with a decrease in cholesteryl ester uptake. ANG II-mediated suppression may contribute to various pathophysiological situations, such as acute tubular injury, the nephrotic syndrome and atherosclerosis. PMID- 15894809 TI - Surviving the pills and the doctor! PMID- 15894810 TI - Correlation between maternal plasma homocysteine and zinc levels in preeclamptic women. AB - Women with preeclampsia have been shown to have elevated blood levels of the metabolite homocysteine, and alterations in blood levels of zinc and copper have also been reported. This study measured plasma levels of zinc, copper, and homocysteine in women with preeclampsia and in women with healthy, normotensive pregnancies. For the patients with preeclampsia compared with controls, significantly higher mean plasma levels were found of homocysteine (16.39 vs 9.45 nmol/mL; p pamidronate. However, coincubation with excess Zn2+ or Mg2+ completely abolished this inhibitory effect. Electrophoretic analysis rendered very similar results: namely a decrease in the enzymatic activity of the bone-ALP band by BPs and a reversion of this inhibition by divalent cations. This study shows that N-containing BPs directly inhibit bone ALP activity, in a concentration range to which this exoenzyme is probably exposed in vivo. In addition, this inhibitory effect is most possibly the result of the chelation of Zn2+ and Mg2+ ions by BPs. PMID- 15894814 TI - Blood pressure relationship to nitric oxide, lipid peroxidation, renal function, and renal blood flow in rats exposed to low lead levels. AB - The results of experiments designed to show that inhibition of nitric oxide production in rats exposed to low lead levels increases vascular resistance, decreases renal blood flow and glomerular function, and enhances oxidative stress. Forty-five adult male Sprague-Dawley rats were divided into four groups. Group A was used as controls and consisted of rats that received no treatment; group B acted as NO-inhibited controls by receiving L-NAME (N(G)-nitro-l-arginine methyl ester) as the NO inhibitor; group C was injected intraperitoneally with 8 mg/kg lead acetate for 2 wk; and group D receiving lead acetate plus L-NAME. Compared to healthy controls, significant elevation of the mean (p<0.01), systolic (p<0.04), and diastolic (p<0.01) blood pressures was found in the lead treated rats. The renal blood flow was 1550+/-468 blood per unit (bpu) in the controls, 488+/-220 bpu in the L-NAME controls, 1050+/-458 bpu in the lead treated group, and 878+/-487 bpu in the Pb plus L-NAME group. Low-level lead exposure did not change the urinary flow rate, creatinine clearance, and the creatinine, potassium, phosphorus, glucose, and protein excretion in 24-h urine. In the lead plus NO-inhibited rats, a significant decrease in sodium ion excretion was observed (p<0.01). The NO levels of the lead exposed, L-NAME treated controls, and L-NAME plus lead-exposed groups are significantly lower compared to untreated controls: p<0.002, p<0.001, and p<0.01, respectively. When compared to untreated controls, the plasma malondialdehyde levels were not significantly different in the lead exposed, lead plus L-NAME, and L-NAME control groups. These results suggest that lead-induced hypertension might be related to a decrease of NO and consequent vasoconstriction, rather than to a decrease of renal blood flow or to decreases in renal sodium. PMID- 15894815 TI - Persistent alterations in biomarkers of oxidative stress resulting from combined in utero and neonatal manganese inhalation. AB - Neonatal female and male rats were exposed to airborne manganese sulfate (MnSO4) during gestation and postnatal d 1-18. Three weeks postexposure, rats were killed and we assessed biochemical end points indicative of oxidative stress in five brain regions: cerebellum, hippocampus, hypothalamus, olfactory bulb, and striatum. Glutamine synthetase (GS) protein levels, metallothionein (MT) and GS mRNA levels, and total glutathione (GSH) levels were determined for all five regions. Overall, there was a statistically significant effect of manganese exposure on decreasing brain GS protein levels (p=0.0061), although only the highest dose of manganese (1 mg Mn/m3) caused a significant increase in GS messenger RNA (mRNA) in both the hypothalamus and olfactory bulb of male rats and a significant decrease in GS mRNA in the striatum of female rats. This highest dose of manganese had no effect on MT mRNA in either males or females; however, the lowest dose (0.05 mg Mn/m3) decreased MT mRNA in the hippocampus, hypothalamus, and striatum in males. The median dose (0.5 mg Mn/m3) led to decreased MT mRNA in the hippocampus and hypothalamus of the males and olfactory bulb of the females. Overall, manganese exposure did not affect total GSH levels, a finding that is contrary to those in our previous studies. Only the cerebellum of manganese-exposed young male rats showed a significant reduction (p<0.05) in total GSH levels compared to control levels. These data reveal that alterations in biomarkers of oxidative stress resulting from in utero and neonatal exposures of airborne manganese remain despite 3 wk of recovery; however, it is important to note that the doses of manganese utilized represent levels that are 100-fold to a 1000-fold higher than the inhalation reference concentration set by the US Environmental Protection Agency. PMID- 15894816 TI - Effect of selenium on expression of selenoproteins in mouse fibrosarcoma cells. AB - Selenium (Se), an essential trace element, is incorporated into selenoproteins as selenocysteine using insertion machinery, including UGA codon and selenocysteine insertion sequence (SECIS) element in the 3'-untranslated region (3'-UTR) of mRNA. To assess the biological effects of tumor cells exposed to the elevated, but nontoxic Se level on glutathione peroxidase (GPx1 [cellular] and GPx3 [extracellular]), thioredoxin reductase (TrxR), and selenoprotein P (SeP) mRNA expression, we introduced a semiquantitative reverse transcription-polymerase chain reaction technique for each selenoprotein transcript using beta-actin as a reference housekeeping gene in mouse fibroblasts (WEHI 164). Cell lines were cultured with 1.0, 2.5, and 5.0 ng of Se in 1 mL of medium for 3 and 7 d, apart from the control cell line with standard medium. It was found that Se exerts a statistically significant (p<0.05) effect only on GPx3 mRNA, referred to as the optical density (OD) ratio (GPx3/beta-actin). Moreover, the lowest Se level affected GPx3 mRNA expression more strongly than its highest concentrations. In an in vitro model applied in this study, GPx3 gene expression is most specific for Se supplementation. PMID- 15894817 TI - In vivo iron and zinc deficiency diminished T- and B-selective mitogen stimulation of murine lymphoid cells through protein kinase C-mediated mechanism. AB - Zinc and iron are crucial mineral components of human diet, because their deficiency leads to several disorders, including alterations of the immune function. It has been demonstrated, in both humans and rodents, that a diminished number of lymphoid cells and a loss of lymphocyte activity accompany deprivation of these essential minerals. The aim of this work was to analyze if iron and/or zinc imbalances regulate lymphocyte activity and the intracellular signals involved in the effect. Mice from the BALB/c strain were fed with iron- and/or zinc-deficient or mineral-supplemented diets, according to the American Institute of Nutrition Rodent Diets. Levels of iron and zinc were assessed in blood, liver, or bone samples. Selective mitogen stimulation of T- and B-lymphocytes were performed. We found a diminished proliferative response in T- and B-lymphocytes from zinc- and/or iron-deficient animals with respect to controls. These effects were related to decreased mitogen-induced translocation of protein kinase C (PKC) activity to cell membranes on both cell types from all animals fed with deficient diets. Our results demonstrate that iron and zinc deficiencies affect both T- and B-lymphocyte function by PKC-dependent mechanisms. PMID- 15894818 TI - Effect of La(III) on the growth and aging of root of loquat plantlet in vitro. AB - The effect of lanthanum on the adventitious root growth, activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD), O2* production rate, MDA content, and the relative plasma membrane permeability in roots of loquat test tube plantlet were studied. Addition of lanthanum chloride (0.5 micromol/L) to the culture medium significantly increased the length of roots, increased the dry weight, enhanced the activities of SOD, CAT, and POD, decreased O2* production rate, malondialdehyde (MDA) contents, and the relative plasma membrane permeability. The relatively stable membrane structure of cell could defer the root aging of the plantlet in vitro. There are important theory meaning and practical value in applying LaCl3 in the rooting medium to raise the rate of rooting and transplant for wood plant. PMID- 15894819 TI - Passive transverse mechanical properties as a function of temperature of rat skeletal muscle in vitro. AB - The objective of the current study was to determine the in vitro passive transverse mechanical properties of skeletal muscle with Dynamic Mechanical Thermal Analysis (DMTA) tests. The starting hypotheses was that the time temperature-superposition principle could be used to expand the DMTA results to a 1 kHz frequency range. Experiments were performed with rat hind leg skeletal muscle tissue samples on a rotational rheometer using a parallel plate geometry. Because of the small size and low modulus of the samples, the standard test geometry was altered and the samples were shifted from the center to the edge of the plates. From strain sweep tests it became clear that for strains smaller than 0.003 the muscle tissue behaves linearly. In the linear region storage moduli ranged between 24 kPa (omega = 1 rad/s) and 42 kPa (omega = 100 rad/s) at T = 4 degrees C and 22 kPa and 33 kPa at 29 degrees C within the experimental frequency range. The loss modulus decreased with increasing frequency and ranged between 7 and 4 kPa at 4 degrees C and 4.5 and 3.5 kPa at 29 degrees C. Although the properties are clearly temperature dependent, a temperature shift in phase angle delta could not be detected, thus Time Temperature Superposition is not allowed for skeletal muscle in vitro. PMID- 15894820 TI - Shear linear behavior of brain tissue over a large frequency range. AB - The literature review about the shear linear properties of brain tissue reveals both a large discrepancy in the existing data and a crucial lack of information at high frequencies associated with traffic road and non-penetrating ballistic impacts. The purpose of this study is to clarify and to complement the linear material characterisation of brain tissue. New data at small strains and high frequencies were obtained from oscillatory experiments. The tests were performed on thin porcine white matter samples (corona radiata) using an original custom designed oscillatory shear testing device. At 37 degrees C, the results showed that the mean storage modulus (G') and the mean loss modulus (G'') increased with the frequency (0.1 to 6310 Hz) from 2.1+/-0.9 kPa to 16.8+/-2.0 kPa and from 0.4+/-0.2 kPa to 18.7+/-2.3 kPa respectively. The reliability of these new dynamic data was checked over a partially common frequency range by conducting similar experiments using a standard rheometer (Bohlin C-VOR 150). Data were also compared in the time field. From these experiments, the relaxation modulus (G(t)) was found to decrease from 24.4+/-2.1 kPa to 1.0+/-0.3 kPa between 10(-5) s and 270 s. PMID- 15894821 TI - Role of Toll-like receptor 4/NF-kappaB pathway in monocyte-endothelial adhesion induced by low shear stress and ox-LDL. AB - TLR4 plays an important role in atherosclerosis, but little is known about the precise mechanism. Herein, we investigated the role of TLR4/NF-kappaB signaling pathway in monocyte-endothelial adhesion induced by low shear stress and Ox-LDL. We found that low shear stress up-regulated TLR4 expression in endothelial cells, and that ox-LDL exerted an obvious synergistic action as revealed by RT-PCR and Western blotting analysis. Low shear stress also significantly up-regulated IL-8 expression in endothelial cells. Meanwhile, NF-kappaB activity and the adhesion force of monocytes were increased, and there was a synergetic action of ox-LDL. However, following transfection with a functional mutant of TLR4 (C3H/HeJ, TLR4 Dicd) or addition of anti-human TLR4 mAb, IL-8 expression was obviously decreased, NF-kappaB activity in cells remarkably inhibited, and the adhesion force of monocyte significantly reduced. Nevertheless, anti-human TLR2 mAb had no similar effects. These findings suggest that TLR4 may be involved in the early stages of atherosclerosis, associating ox-LDL, inflammation/infection, and low shear stress. Therefore, TLR4 is expected to be a new target for preventing and treating atherosclerosis. PMID- 15894822 TI - An automated tube-type blood viscometer: validation studies. AB - The technical complexity of previous rheometers has tended to limit the availability of blood viscosity data obtained over a wide range of shear rates. However, an automated tube-type viscometer, the Rheolog, has been developed; it employs a disposable flow assembly and less than five minutes are required to obtain blood viscosity results over a shear rate range of 1-1500 s(-1). We have carried out validation studies of the Rheolog using normal human blood and have compared these results with those obtained by cone-plate and Couette viscometers; storage time and temperature effects were also evaluated. Replicate measurements indicated mean CV levels less than 5%, and were independent of hematocrit and shear rate. Rheolog blood viscosity data agreed closely with those from other viscometers: average Rheolog differences from mean cone-plate and Couette values were -0.3% at 28% hematocrit, -1.4% at 41% hematocrit (i.e., native), and 1.0% at 56% hematocrit. Storage at room temperature up to 8 hours and at 4 degrees C up to 4 days had minimal effects whereas notable changes were observed when stored for 3 hours at 37 degrees C. Our results indicate that, within the hematocrit and shear rate limits employed herein, the Rheolog provides rapid, accurate and reproducible blood viscosity data, and suggest its usefulness for both basic science and clinical studies. PMID- 15894823 TI - Cell morphological changes in venous remodeling induced by arteriovenous grafting in rat limb. AB - Vascular remodeling induced in rat limb by arteriovenous (AV) shunting was investigated by evaluating changes in vascular diameter and cell morphology. In Wistar rats, a vein graft was implanted in situ in the hind limb. Flow-rate in the grafted vein was assessed by measuring flow in the common femoral artery using an ultrasonic flowmeter. Nuclei and actin filaments of the venous wall were stained with propidium iodine and phalloidine-FITC, and the samples were observed using confocal laser microscopy. The grafted veins became circular in cross section with increase in diameter during two weeks after AV shunting. Owing to the increase in diameter, the estimated wall shear stress was not increased so much as the flow-rate. The confocal laser microscopic observation showed that endothelial cells (ECs) and smooth muscle cells (SMCs) in the grafted veins were either aligned well (2 out of 8 samples), or ECs were denudated and SMCs were disrupted (in 6 out of 8 samples). The cell density of ECs was unchanged from the control level. In conclusion, the grafted vein was remodeled with morphological changes in ECs and SMCs during 2 weeks after AV shunting. PMID- 15894824 TI - Adhesion of monocyte-derived dendritic cells to human umbilical vein endothelial cells in flow field decreases upon maturation. AB - Dendritic cells (DC) are sentinels of the immune system. They and their precursors undergo complex migration to perform their function in vivo. Binding of DC to vascular endothelial cells in a flow field has not been investigated. We therefore determined adhesion of DC and their precursors, MO, to human umbilical vein endothelial cells (HUVECs) under various shear stresses by using a flow chamber system. The results showed that the binding was reduced with developmental stages of DC, which partially depended on CD11a and cell surface charges. The data had potential relevance for anti-cancer immunotherapy strategies favoring the application of mDC. PMID- 15894825 TI - Analysis of aggregation parameters of erythrocytes in diabetes mellitus. AB - Aggregation properties of normal and diabetic subjects were analyzed by optical scattering under dynamic conditions. Aggregation of erythrocytes was determined using on-line erythrocyte aggregometer based on sequential analysis of transmitted laser light intensity after passing through the erythrocyte suspension. Diabetes samples were divided into four groups depending upon their blood glucose levels. The aggregation mechanism was determined in terms of aggregation size index, effective number of cells and effective cellular sedimentation duration. These parameters thus obtained were used to quantitatively describe the dynamic nature of aggregation process under gravitational sedimentation. The aggregation parameters decreased in diabetic samples. This indicates that the aggregation of erythrocytes was significantly increased in diabetes mellitus. These changes may affect flow properties of erythrocytes in the microcirculation. PMID- 15894826 TI - Vasodilatation of human gingiva and neurogenic inflammation. AB - BACKGROUND: Neurogenic Inflammation (NI) is the consequence of amyelinic sensitive neuron activation. Recent studies on rats proved that NI could be experimentally induced by topical capsaicin application. The aim of this study is to evaluate the effects of topical capsaicin application on human periodontal mucosa and to assess if NI might have a role in the pathogenesis of periodontal diseases. METHODS: 15 patients were examined in our laboratory. NI was experimentally induced in the gingival mucosa close to: (1) the interdental papilla corresponding to the upper central incisors; (2) the interdental papilla corresponding to the lower left lateral incisor and canine after ipsilateral nerve trunk anaesthesia. The characteristics of gingival microcirculation were observed using computerised videocapillaroscopic techniques. RESULTS: axon reflected vasodilatation was observed close to the papilla corresponding to the upper central incisors. An important correlation was observed between capsaicin application and capillary tortuosity. No significant modifications in vascular diameter and tortuosity were observed after capsaicin application close to the interdental papilla corresponding to the lower left lateral incisor and canine. CONCLUSIONS: the study shows that NI can be induced in human gingiva, and such evidence could be extremely important in the pathogenesis and treatment of periodontal diseases. PMID- 15894827 TI - Serum C-reactive protein and circulating endothelial cells in patients with acute myocardial infarction. AB - As an in vivo indicator of vascular injuries, circulating endothelial cells (CECs) have been used as a marker of endothelial damage in a variety of vascular disorders. C-reactive protein (CRP) is a sensitive indicator of inflammation. To investigate the presence of CECs in patients with acute myocardial infarction (AMI) and to evaluate their clinical associations and possible relationship with infection, CECs from peripheral blood were isolated by using immunomagnetic beads coated with antibodies against CD146. Their endothelial origin was confirmed by the positive labeling of von Willebrand Factor (vWF), CD31 and electron microscope. Results showed that CECs number and CRP level were 52 (28-81.5) cell/ml, 7.98+/-2.25 mg/l and 10.5 (6-16.5) cell/ml, 1.41+/-2.05 mg/l in AMI patients (n=37) and health controls (n=42) respectively (p<0.001). The apoptosis rate and necrotic rate of CECs in AMI were 25% and 19%, respectively. Correlation analysis revealed a significant positive correlation between CECs and CRP (r=0.505, p=0.001). CECs numbers did not correlate with age, gender, serum cholesterol, hypertension, obesity, history of cardiovascular disease, or smoking. These results suggest a strong correlation between level of CRP and counts of CECs in patients with AMI. The number of CECs and CRP may be combined to reflecting the endothelial cell injury. PMID- 15894828 TI - Effect of low-intensity aerobic exercise training on arterial compliance in postmenopausal women. AB - Regular aerobic exercise training attenuates age-related reduction in central arterial compliance, an independent risk factor of cardiovascular diseases. We tested the hypothesis that even low-intensity exercise training could increase central arterial compliance in postmenopausal women. Using B-mode ultrasound, we studied the central arterial compliance of 15 postmenopausal females (age: 52-66 years) before and after a 12-week aerobic exercise intervention. Subjects performed aerobic exercise training of the same energy expenditure (cycle exercise, total 900 kcal/week, 3-5 sessions/week) at two different exercise intensities: 7 trained at low intensity (40% heart rate reserve: L-TR) and 8 trained at moderate intensity (70% heart rate reserve: M-TR). Arterial compliance increased after exercise training in the L-TR group (0.70+/-0.32 vs. 1.06+/-0.55 mm2/mmHgX10(-1), p <0.05) and in the M-TR group (0.82+/-0.37 vs. 1.14+/-0.39 mm2/mmHgX10(-1), p <0.05). There was no significant difference in increases of arterial compliance in either group (L-TR: 0.35+/-0.38 vs. M-TR: 0.32+/-0.33 mm2/mmHgX10(-1)). These results suggest that the improvement of central arterial compliance by aerobic exercise training might not be influenced by the intensity of exercise training if the energy expenditure of the training is the same. Accordingly, even low-intensity exercise training may have the effect of improving central arterial compliance. PMID- 15894829 TI - Association study using single nucleotide polymorphisms in the estrogen receptor beta (ESR2) gene for preeclampsia. AB - The pathophysiology of preeclampsia (PE) remains uncertain despite many research efforts. Actual hypotheses seek to explain the vascular damage that characterizes the disease. Recently, it was reported that the mouse disrupted estrogen receptor beta (ESR2) gene was associated with abnormal vascular function and hypertension. Moreover, some investigators have reported that subjects with a family history of hypertension have a statistically significant increased risk for PE. Thus, it is thought that the pathophysiology of PE overlaps that for hypertension. The aim of the present study was to investigate the relationships between single nucleotide polymorphisms (SNPs) in the human ESR2 gene and PE in Japanese subjects, and to assess the involvement of a family history of hypertension in these relationships. Based on a database search on the web site of the National Center of Biotechnology Information (NCBI), we chose four SNPs in the human ESR2 gene, and performed an association study in 84 PE patients and 160 age-matched non-PE subjects. The overall distribution in each SNP did not differ significantly between the two groups. However, after dividing the groups into subjects with and without a family history of hypertension, the allelic distribution of one of the SNPs (rs928554) revealed a positive association. Thus, a possible mutation linked to a SNP may prescribe a genetic predisposition for patients with a family history of hypertension in PE. PMID- 15894830 TI - A consistent abnormality in the average local smoothness of fetal heart rate in growth-restricted fetuses affected by severe pre-eclampsia. AB - An abnormality in cardiovascular regulation during the prenatal period has been suggested to be the pathophysiological link between fetal growth restriction and adult hypertension. The purpose of this study was to determine how consistently abnormal the local smoothness of the very-short-term heart rate is in growth restricted fetuses associated with severe pre-eclamptic pregnancy. Multifractal Hurst analysis on the structure function of heart rate was performed in control fetuses (n =150), in fetuses affected by severe pre-eclampsia and not showing growth restriction (n =66) and in fetuses affected by severe pre-eclampsia and showing growth restriction (n =58). The very-short-term (< or =15 heart beats) generalized Hurst exponents of the order of -5 to 5 in three groups were compared. Each exponent quantifies an average local heart rate smoothness at 15 successive-heart rate sites, which were specified by the magnitude of the heart rate variation within the sites determined by and positively correlated with the order of the exponent. This means that the fetal heart rates within the sites of q > or =2 have a large fetal heart rate (FHR) variation, and those within the sites of q < or =-2 have a small FHR variation. In the fetuses affected by severe pre-eclampsia and not showing growth restriction, only values of the exponents of the order > or =2 were abnormally lower. In the fetuses affected by severe pre eclampsia and showing growth restriction, the values of the exponents of all orders were abnormally lower. In conclusion, the local smoothness of heart rate is consistently abnormal regardless of the magnitude of heart rate variation within a very-short-term period in growth-restricted fetuses affected by severe pre-eclampsia. PMID- 15894831 TI - Association of a GNAS1 gene variant with hypertension and diabetes mellitus. AB - Previous studies have shown that the T allele of the GNAS1 T393C polymorphism is associated with poor responsiveness to beta-blockade and that the T393C polymorphism interacts with cigarette smoking and alcohol consumption in the pathogenesis of hypertension. Thus, the T393C polymorphism is likely to interact with beta-adrenoceptor (beta-AR) stimulation in the pathogenesis of hypertension. Although this interaction might be caused by a direct effect of Gs proteins on the cardiovascular system, it could also result from an indirect effect of Gs proteins mediated by glucose metabolism. Moreover, association studies are often irreproducible. We therefore examined the possible interaction between the T393C polymorphism and gamma-glutamyl transpeptidase (GGT), which is an established biomarker of alcohol consumption, in the association with glucose metabolism as well as with hypertension in a Japanese population. Genotyping for GNAS1 was performed by using the polymerase chain reaction-restriction fragment length polymorphism method in all 821 samples. The present study showed a significant interaction between the T393C polymorphism and GGT in the association with hypertension (p =0.033). This interaction was even more significant after adjustment for all confounding factors (p =0.0025). In contrast, analysis of the possible interaction of the T393C polymorphism with GGT in the association with diabetes mellitus or fasting plasma glucose failed to show a significant result. These results did not support the hypothesis that the interaction between the T393C polymorphism and GGT in the association with hypertension could be caused by an indirect effect of Gs proteins mediated by glucose metabolism. PMID- 15894832 TI - Relationship between brachial-ankle pulse wave velocity and heart rate variability in young Japanese men. AB - This study examined the relationship between arterial stiffness and autonomic nervous function in a young population. A cross-sectional study was conducted on 382 Japanese males, aged 24 to 39 years, who worked at the same information service company. Brachial-ankle pulse wave velocity (baPWV) was measured using an automatic waveform analyzer, and the spectral power of heart rate variability in the low frequency (LF: 0.04-0.15 Hz) and the high frequency (HF: 0.15-0.40 Hz) band was evaluated by the maximum entropy method. LF/HF and HF were used as the indicators of sympathetic and parasympathetic nervous activity, respectively. Psycho-hormonal responses were examined by the Profile of Mood State (tension anxiety and anger-hostility scales) and Hamilton's Depression Scale with serum cortisol and catecholamine levels. In a univariate analysis, baPWV was positively associated with the following variables (all p <0.05): LF/HF, age, body mass index, systolic and diastolic blood pressures, heart rate, serum total cholesterol and triglycerides, blood glucose, and plasma cortisol and noradrenaline. Multiple regression analysis indicated that LF/HF was an independent predictor of baPWV (p <0.05), after controlling for significant effects of age, systolic blood pressure, and plasma noradrenaline levels. There was no significant effect of HF on baPWV in this multivariate analysis. Neither mood state nor health-related lifestyle factors such as smoking were significant. It was suggested that baPWV is closely associated with sympathetic nervous activity in young men. PMID- 15894833 TI - Relationship of bradykinin B2 receptor gene polymorphism with essential hypertension and left ventricular hypertrophy. AB - The bradykinin B2 receptor shows a protective role in the development of hypertension and renal and cardiovascular complications. It was recently reported that a polymorphism of the bradykinin B2 receptor gene (BDKRB2) is a genetic predisposing factor for hypertension and cardiovascular disease. The aim of this study was to examine the relationship of a polymorphism (-58 T/C and exon 1 +9/ 9) of BDKRB2, and an insertion/deletion polymorphism (I/D) of the angiotensin converting enzyme gene (ACE) with essential hypertension and cardiovascular mortality in the Japanese population. Genotyping was carried out in 275 hypertensive and 441 normotensive subjects. Left ventricular hypertrophy (LVH) was detected by ECG in 242 untreated patients with hypertension. All participants were Japanese and gave their written informed consent. The polymorphism (-58 T/C) in the promoter region of the BDKRB2 was determined using the TaqMan-polymerase chain reaction (PCR) method, the exon 1 +9/-9 polymorphism of the BDKRB2 and I/D polymorphism of the ACE were monitored by PCR and gel electrophoresis. The genotypes and allelic frequencies were in Hardy-Weinberg equilibrium. The polymorphism (-58 T/C) in the promoter of the BDKRB2 was associated with LVH in the hypertensive group (n =242) (p =0.048; chi2 =3.9; odds ratio: 1.8; 95% confidence interval (CI): 1.0-3.3). Furthermore, the frequency of LVH in hypertensives was significantly higher in the subjects with both the BDKRB2 CC and ACE D allele than those with other genotypes (p =0.002, chi2 =9.4). However, no relationship could be found between polymorphism of the BDKRB2 (p =0.86, chi2 =0.3) or the ACE (p =0.21, chi2 =3.1) and hypertension in this group of subjects. These results suggest that the polymorphism (-58 T/C) in the promoter region of BDKRB might be a risk factor and might have a synergetic effect with the ACE for LVH in hypertensives, but it is not associated with hypertension in the Japanese population. PMID- 15894834 TI - Morning rise in blood pressure is a predictor of left ventricular hypertrophy in treated hypertensive patients. AB - To assess the relationship between home blood pressure and left ventricular mass, we evaluated cardiac echocardiography in 297 hypertensive subjects (188 men and 109 women; mean age, 62.8+/-10.3 years) who were treated with amlodipine monotherapy over 1 year (mean dose, 5.5+/-2.3 mg/day). The morning hypertension group (n=57; 19.2%), who had a morning home systolic blood pressure (HSBP) > or =135 mmHg and an evening HSBP <135 mmHg, had a significantly greater left ventricular mass index (LVMI) concomitant with an increase in the homeostasis model assessment insulin resistance index (HOMA-IR) compared to the good control group (n=174; 58.6%), whose morning and evening HSBP were both <135 mmHg, and had a LVMI roughly equivalent to that of the poor control group (n=63; 21.2%), whose morning and evening HSBP were both > or =135 mmHg. By grouping of subjects according to the difference between morning and evening HSBP (delta HSBP), subjects with a delta HSBP> or =10 mmHg had a significantly greater LVMI than subjects with a delta HSBP <10 mmHg. Increases in LVMI in these patients were still significant after adjustment for age, gender, dose of amlodipine, alcohol consumption, body mass index, office systolic blood pressure, and morning and evening HSBP. In a stepwise multivariate regression analysis, delta HSBP (r2=36.2%, p <0.001), morning HSBP (r2=5.5%, p <0.001), HOMA-IR (r2=1.4%, p=0.016) and age (r2=1.0%, p=0.026) were determined to be significant contributing factors for LVMI. This regression model could explain 44.1% of LVMI variability. These results suggest that morning rise in blood pressure is a dominant predictor of left ventricular hypertrophy. PMID- 15894835 TI - Moderate regular exercise increases basal production of nitric oxide in elderly women. AB - Vascular endothelial cells produce nitric oxide (NO), which is a potent vasodilator substance and is thought to have antiatherosclerotic properties. Therefore, it has also been proposed that NO may be useful to regulate vascular tonus and prevent progression of atherosclerosis. On the other hand, NO activity reduces with aging. We previously reported that the plasma nitrite/nitrate (NOx: the stable end product of NO) concentration was significantly increased by intense aerobic exercise training in healthy young humans. We hypothesized that lifestyle modification (e.g., even mild regular exercise training) can increase NO production in previously sedentary older humans. We measured the plasma NOx concentration before and after a mild aerobic exercise training regimen (cycling on a leg ergometer at 80% ventilatory threshold for 30 min, 5 days/week) for 3 months in elderly women. In addition, we assessed the plasma concentration of cyclic guanosine monophosphate (cGMP), a second messenger of NO, in the same samples. The individual ventilatory threshold increased significantly after the 3 month exercise training. The blood pressure at rest significantly decreased after exercise training. These results suggest that the 3-month exercise training in the older women produced favorable physiological effects. The plasma concentration of NOx significantly increased by the exercise training, and the plasma concentration of cGMP also increased by the exercise training. The present study suggests that even a mild regular aerobic-endurance exercise increases NO production in previously sedentary older humans, which may have beneficial effects (i.e., antihypertensive and antiatherosclerotic effects by endogenous NO) on the cardiovascular system. PMID- 15894836 TI - C-reactive protein and peripheral artery disease among Japanese elderly: the Tsurugaya Project. AB - We investigated the cross-sectional relationship between ankle brachial index and cardiovascular disease risk factors, including C-reactive protein (CRP), among Japanese elderly, a topic which has had little prior epidemiologic study. Our study population comprised 946 subjects aged at least 70 years in whom both CRP and ankle brachial index were measured. The participants were classified into a low (ankle brachial index<0.9) and normal ankle brachial index group. We found that current smoking, high-density lipoprotein cholesterol <40 mg/dl, a low body mass index (continuous variable), hypertension, diabetes and statin use were all significantly related to a lower ankle brachial index. Higher log-transformed CRP level was significantly related to a lower ankle brachial index after adjustment for the cardiovascular risk factors mentioned above (p <0.01). The odds ratios for low ankle brachial index compared to 0-1 risk factors were 5.79 (95% confidence interval [CI]: 2.99-11.20) for 2 risk factors and 17.45 (95% CI: 6.78 49.91) for 3 or more risk factors; independently of other risk factors, the odds ratio for CRP>1.0 mg/l was 2.10 (95% CI: 1.13-3.88) compared to lower CRP values. Thus, a high level of CRP is related to a low ankle brachial index among Japanese elderly as well as Western subjects. This is the first study to report the relationship between CRP and low ankle brachial index among Japanese elderly. PMID- 15894837 TI - Low-dose combination therapy with temocapril and losartan reduces proteinuria in normotensive patients with immunoglobulin a nephropathy. AB - This study investigates the ability of low doses of angiotensin-converting-enzyme inhibitors, in combination with angiotensin II receptor blockers, to exert antiproteinuric effects in normotensive and proteinuric outpatients with immunoglobulin A (IgA) nephropathy confirmed by biopsy. We performed a prospective, randomized, 6-month study of the effects of temocapril 1 mg (n=10), losartan 12.5 mg (n=10), and both (n=11) on mild-to-moderate proteinuria 0.76+/ 0.35 g/day (range, 0.4 to 1.6 g/day) and renal function. The study subjects comprised 31 normotensive and proteinuric outpatients with IgA nephropathy accompanied by normal, or mild-to-moderately reduced but stable renal function (glomerular filtration rate>50 ml/min) without steroid or immunosuppressive therapy. We prospectively evaluated blood pressure, proteinuria, renal function and biochemical parameters before and after 6 months of therapy. The combination therapy significantly reduced proteinuria (63.2%) compared with either temocapril or losartan alone (41.3% and 36.6%, respectively, p=0.04 and 0.01, respectively). Blood pressure was most decreased in the group that received combination therapy. The reduced proteinuria did not correlate with reduced systolic or diastolic blood pressure or mean arterial pressure in any of the groups. The glomerular filtration rate fell during the first 3 months of combined therapy, but became reversible after a further 3 months of therapy. The combination significantly decreased angiotensin II (p <0.01), and this decrease was greater than that by either drug alone. In conclusion, the effectiveness of the combined therapy may have been at least partly due to the greater inhibition of the action of angiotensin II in patients with IgA nephropathy. This strategy apparently reduced mild-to-moderate proteinuria in patients with normotensive IgA nephropathy. PMID- 15894838 TI - Spironolactone in combination with cilazapril ameliorates proteinuria and renal interstitial fibrosis in rats with anti-Thy-1 irreversible nephritis. AB - Blockade of the renin-angiotensin system has been established as a treatment for heart failure with hypertension and left ventricular hypertrophy, and for progressive kidney diseases. The present study was conducted to examine whether spironolactone, a mineralocorticoid receptor antagonist, alone or in combination with cilazapril, an angiotensin converting enzyme (ACE) inhibitor, ameliorates proteinuria and renal lesions in an immune-initiated progressive nephritis model. Wistar rats were uninephrectomized 7 days before injection of anti-Thy-1 monoclonal antibody 1-22-3 to induce progressive glomerulonephritis. The nephritic rats were untreated or treated with spironolactone (400 mg/kg body weight/day), cilazapril (1 mg/kg body weight/day), or both for 10 weeks. Proteinuria was increased in the untreated rats 1 week after nephritis induction and was maintained throughout the experiment. Compared with the untreated animals (212.9+/-49.2 mg/day), proteinuria was significantly reduced in the spironolactone-treated group (62.0+/-4.0 mg/day, p=0.0046) and the cilazapril treated group (71.8+/-26.0 mg/day, p=0.0048) on day 70 after antibody injection. Further reduction of proteinuria (42.4+/-4.5 mg/day, p=0.0019 vs. the untreated group) and less renal cortex interstitial fibrotic change (fibrosis score: 142.0+/-18.4 vs. 80.3+/-18.5 in the untreated group, p=0.0123) were detected in the spironolactone plus cilazapril-treated group. Blood pressure did not differ among the three treatment groups. In conclusion, spironolactone ameliorates proteinuria to the same degree as cilazapril, and concomitant use of spironolactone and an ACE inhibitor further suppresses renal disease progression. These data suggest that concomitant treatment with spironolactone and an ACE inhibitor has beneficial effects on immune-initiated progressive kidney disease. PMID- 15894839 TI - A serine protease inhibitor, nafamostat mesilate, suppresses aldosterone secretions in vivo. AB - To examine the role of serine proteases in the control of aldosterone (Ald) secretion, we studied the effects of nafamostat mesilate (Naf), a serine protease inhibitor, on in vivo Ald secretion and Ald content in the rat adrenal gland. Either Naf (2 mg/kg/h; n=10) or saline (2 ml/h; n=10) was administered intravenously for 30 min to anesthetized Wistar rats whose left adrenal vein was cannulated selectively via the inferior vena cava. Naf caused a significant decrease in Ald secretion rate compared to saline (1.99+/-0.32 vs. 3.42+/-0.56 ng/min, p <0.001), while adrenal blood flow, mean arterial pressure and plasma renin activity in the adrenal venous blood did not differ between the two groups. In a separate trial, adrenal Ald content, adrenal renin content, plasma adrenocorticotropic hormone (ACTH) and plasma potassium did not differ between rats treated with Naf (n=7) and those administered saline (n=7). These data suggested that Naf-inhibitable serine proteases may participate in the control of Ald secretion through mechanism(s) other than hemodynamic changes, adrenal renin, ACTH, and/or plasma potassium. PMID- 15894840 TI - Two cases of renovascular hypertension and ischemic renal dysfunction: reliable choice of examinations and treatments. AB - We experienced two aged patients with atherosclerotic renovascular stenosis associated with hypertension and ischemic nephropathy. Both patients exhibited sudden rise in blood pressure (BP) and progressive aggravation of renal dysfunction. In these patients, the use of contrast medium to screen for renal artery stenosis (RAS) ran the risk of further deterioration of renal function. We therefore used magnetic resonance angiography (MRA), which is less conducive to renal damage, to screen for RAS. One-sided RAS was treated by percutaneous transluminal angioplasty of the renal artery (PTRA) and stenting. As a result, BP decreased in both patients. Serum creatinine (Cr) decreased slightly in one patient, whereas, in the other, serum Cr increased transiently and then decreased and stabilized to pre-treatment levels. Thus, although it is unclear whether the combination of PTRA and stenting is among the best treatments for patients with RAS and moderate-to-severe renal dysfunction, PTRA and stenting are clearly of benefit in selected patients. In addition, recent progress in characterizing the pathophysiology of ischemic nephropathy associated with renovascular hypertension has created interest in the therapeutic potential of angiotensin II receptor antagonists, sympatholytic agents, and antioxidants. Therefore, we discuss the therapeutic utility of PTRA and stenting and the above-mentioned medications in patients with RAS and renal dysfunction. PMID- 15894841 TI - Histopathological findings in 851 autopsies of drug addicts, with toxicologic and virologic correlations. AB - This investigation was carried out on 851 consecutive judicial autopsies of drug addicts who died mostly of heroin overdose from 1977 to 1996. Research of anti HIV/HBV/HCV antibodies was performed, and histologic sections were retrospectively reviewed. More than 65% were HBV/HCV-positive and about 17% HIV positive; females were HIV-positive more often than males. Intracranial microhemorrhages were frequently found; cerebral infectious diseases were rare. Inflammatory heart lesions, myocardial fibrosis, and acute ischemia were common. Interstitial nephritis (found in about 8%) was more frequent in females, in older patients, and in those carrying HIV infection; glomerular sclerosis was detected in about 12%. Acute bronchitis and/or pneumonia was demonstrated in 12%, without significant association with HIV infection; pulmonary hemorrhages, foreign body granulomas, and food aspiration were also commonly seen; hyperplasia of pulmonary perivascular lymphatic tissue was rather characteristic. Liver was carrying steatosis in 66.3% and/or hepatitis in 64.5%; acute hepatitis was more frequent in females, chronic hepatitis in older subjects and in those proven positive for hepatotropic viruses; cirrhosis occurred more often in older patients, in those carrying virus infection, and in consumers of nonnarcotics drugs such as ethanol. No pathologic finding was clearly related to drug abuse; therefore, during autopsy, drug addiction can be suspected, but anamnestic and circumstantial data are needed to lead pathologists to request toxicologic analysis to ascertain the cause of death. The present investigation emphasizes that, in addition to the risk of death by overdose, the high incidence of acute and chronic diseases could seriously undermine the health status of heroin and/or other drug consumers. PMID- 15894842 TI - Effect of wrist restraint on maximal exercise capacity in healthy volunteers. AB - Each year, reports occur of deaths in individuals, while struggling against physical restraint. The mechanism of these deaths remains unclear. This study aimed to determine the effect of wrist restraint on cardiorespiratory function during maximal exercise. Twelve healthy volunteers underwent 3 incremental maximal exercise tests on a cycle ergometer. In a randomized order, they exercised while unrestrained, with the wrists tied in front of the body or wrists tied behind the back. The primary outcome measures were the number of minutes exercised in each position and heart rate and whole blood lactate level for each stage and on reaching maximum exercise capacity. The mean exercise duration was 19 minutes 6 seconds unrestrained (95% confidence interval [CI] 16 minutes 52 seconds to 20 minutes 57 seconds), 18 minutes 51 seconds (95% CI 17 minutes 51 seconds to 20 minutes 50 seconds) with arms restrained in front and 16 minutes 51 seconds (95% CI 14 minutes 6 seconds to 19 minutes 20 seconds) with the arms restrained behind the body (P = 0.16). There was no significant difference in heart rate or lactate measurements. It is probable that other factors make a more important contribution than wrist restraint behind the body to cardiorespiratory compromise in a struggling and fatigued individual. PMID- 15894843 TI - Lethal injuries occurring during illegal break-ins. AB - Individuals who undertake criminal activity are often at risk of injury for a variety of reasons. Two cases are reported where individuals who were engaged in illegal breaking and entering activities sustained stab wounds from broken window glass and died of exsanguination. The cases both involved young males who were under the influence of alcohol and/or drugs and who sustained relatively minor tissue damage that had fatal consequences due to a variety of coincident factors. Critical location of wound sites over major vessels resulted in rapid bleeding that was enhanced by vasodilator effects of alcohol and stimulant effects of amphetamines. In addition, adrenergic effects of being engaged in illegal activities, and the subsequent pain from the wounds, may also have contributed to accelerated heart rate and blood loss. Failure to appreciate the potential effects of such wounds, which may initially have appeared relatively minor based on their small size, added to an impaired ability to apply first aid or seek appropriate medical assistance, may also have been due to the combined effects of drugs, alcohol, and concerns regarding the illegal nature of the underlying activity. Such a scenario constitutes a specific subgroup of offender injury that could be termed the break, enter, and die syndrome. PMID- 15894844 TI - Forensic medical examination of refugees who claim to have been tortured. AB - The United Nations Convention against torture prohibits repatriation of refugees if there is reason to believe they will be tortured on return to their country. A history of torture is therefore an important factor in making a case for asylum. In this study, the results of the medical examinations of 59 torture victims investigated at the Department of Forensic Medicine, University of Aarhus, Denmark, 1996-2002, are presented and discussed. Variables including age, sex, education, health, torture methods, condition of confinement, torture aftereffects, and findings at the forensic examination were registered in a database (Paradox) and analyzed. In 70%, aftereffects of torture could be documented. These included scars after fixation, burns, incisions, or flogging. Symptoms and signs from joints, muscles, and nerves were common in victims who had been suspended. Many of the victims of phalanga had painful feet and signs of walking impairment. A majority of the victims suffered from posttraumatic stress syndrome. An independent medical report offers an opportunity to evaluate and elaborate the story told by the victim and should be used in disputed cases. We have a duty to protect refugees against torture. PMID- 15894845 TI - The lingual tonsillar hyperplasia in relation to unanticipated difficult intubation: is there any relationship between lingual tonsillar hyperplasia and tonsillectomy? AB - The lingual tonsillar hyperplasia (LTH) is a rare and dangerous condition of the upper airway because it is one of several risks during intubation which can cause serious complications like cardiac arrest, cerebral anoxia, and death. The LTH is rarely detectable during a routine oropharyngeal examination. The lingual tonsil is of particular interest to forensic medicine to assess cases with malpractice in connection with intubation difficulties. Our study looked at how often autopsy specimens had enlarged lingual tonsils and whether any relationship existed between LTH and tonsillectomies. In total, 497 corpses were examined. Only in 16 cases (3.2%), an enlarged lingual tonsil was detected. Of the 16 cases, 10 (62.5%) had normal palatine tonsils; the other 6 cases (37.5%) had no palatine tonsils or were scarred, atrophied, or only remnants remained. Statistically, it was not possible to prove any relationship between enlarged lingual tonsils and tonsillectomies. From the forensic point of view, a description of the lingual tonsil should appear in special cases in autopsy reports. When reviewing malpractice cases, special anatomic features should be described. PMID- 15894846 TI - Bite motivation of sharks reflected by the wound structure on humans. AB - Bite wounds on humans have rarely been comparatively analyzed, and the behavior leading to such bites is virtually unknown. Nevertheless, the behavior of a shark is reflected in the bite structure and should be an essential part of shark accident analysis. This paper compares 3 nonfatal accidents on humans, caused by bull sharks, Carcharhinus leucas, that occurred within a 12-month period in the same area of the Bahamas. Examination focused on wound analysis and accident reconstruction to determine the most likely bite motivation of the sharks. Two sharks targeted the left calf areas of the victims; another one bit the back area of a person. Although both calf bites had a very similar appearance, examination concluded that one of them showed the same triggering behavior as for the shark who inflicted the very different-looking back bite. Those 2 bites were competitive, whereas the other calf bite was initially of exploratory nature, turning into a stress-related bite. PMID- 15894847 TI - Homicide in Adana, Turkey: a 5-year review. AB - Violence is a significant public health problem. Thus, so as to prevent this problem, homicide, the severest form of violence depriving a human being of his right to live, deserves a detailed examination. This study is a retrospective research examining the 2951 cases of medicolegal autopsies in Adana during a period of 5 years (1997-2001). Among these cases, 620, which were determined to be homicidal, were taken into the scope of this study. The cases were examined with respect to sex, age groups, the method used during the act of homicide, the number and the localization of the wounds on the body. A total of 620 (21%) of the medicolegal autopsies conducted within this period were homicides. Of these cases, 515 (83.06%) were male and 105 (16.94%) female, and the rate of the males to females was 4.9; 72.74% of the victims were between the ages of 21 and 50. It was seen that 54.83% of the homicides involved firearms, while 35.16% of the victims were stabbed to death with a cutting object. It was also determined that the victims suffered a single wound in 47.35% of firearm-related murders and 29.35% of stabbings resulted in death. Alcohol was found in the blood of 7.58% of the homicide victims, while none had any illicit drugs. PMID- 15894848 TI - Vitreous humor biochemical constituents: evaluation of between-eye differences. AB - The present study was conducted to investigate the differences in the vitreous humor biochemical concentrations for vitreous electrolytes and calcium in the same pair of eyes at identical postmortem interval (PMI). The vitreous humor samples were collected independently in both eyes from 48 autopsies (PMI range, 4.5-84.3 hours) with documented time of death. The samples were analyzed for potassium, sodium, chloride, and calcium using a Beckman Coulter LX20 Automated Analyzer based on ion-selective electrode methodology. There were no statistically significant between-eye differences at identical postmortem interval. A significantly high correlation was observed between paired potassium concentrations of both the eyes. A highly significant linear correlation was observed between the individual eye and mean potassium concentrations of both the eyes with postmortem interval. The observed differences were not significantly correlated with postmortem interval. The results demonstrated that the between eye differences for vitreous electrolytes and calcium are insignificant. Therefore, the utility of vitreous biochemistry, particularly potassium in postmortem interval estimation and other forensic applications, cannot be questioned solely on the basis of these differences. PMID- 15894849 TI - Vertebral artery laceration mimicking elder abuse. AB - Elder abuse was first described almost 30 years ago. Today, approximately 1 in 25 elders is abused each year in the United States. A newly described form of domestic violence, the incidence of elder abuse will surely increase as the elderly population grows. Physical abuse/inflicted trauma is generally considered the most extreme form of elder mistreatment and includes blunt trauma, sexual assault, traumatic alopecia, and burns. Elder homicide is usually due to gunshot wounds, blunt trauma, stab wounds, or asphyxia. However, the difficult aspect of assessing the possible elder abuse homicide victim is delineating such inflicted trauma from accidental trauma. We report the case of a 94-year-old "demented" male, who reportedly fell out of his wheelchair. He was transported to a local emergency room, where he became unresponsive during examination. He experienced respiratory distress and was pronounced dead shortly thereafter. At autopsy, he had periorbital contusions and a midline abrasion between the eyes, with underlying supraorbital contusion. The skull, brain, and spinal cord were unremarkable for signs of trauma. The major traumatic finding was in the neck region. Neck dissection revealed hemorrhage extending from the base of the skull to the level of T-1 and anteriorly about the soft tissues, strap muscles, and vasculature. The strap muscles were individually examined and were free of hemorrhage. The carotid arteries and jugular veins were unremarkable. The larynx, hyoid, and thyroid were intact, with only surrounding hemorrhage. Further examination revealed a horizontal fracture of the C5 vertebral body and a medial laceration of the left vertebral artery at the C5 level; subarachnoid hemorrhage was absent. What initially appeared to be trauma to the neck, worrisome for strangulation or blunt force trauma, was a large retropharyngeal hematoma from the left vertebral artery laceration. Traumatic rupture of the vertebral artery usually occurs at the C1 and C2 levels, with resultant subarachnoid hemorrhage. This is an especially vulnerable location since it is where the artery turns and then enters the skull. Associated injuries include spinal cord transection or contusion, brachial plexus injury, pharyngoesophageal injury, and vertebral fractures. Retropharyngeal hemorrhage may result from deep neck infection, tumor, and trauma. Hemorrhage associated with trauma often involves flexion of the cervical spine, followed by hyperextension. The accumulation of blood slowly impinges on the pharynx/larynx and vasculature structures. The exact injuries and etiology of the hemorrhage must be determined to distinguish strangulation from blunt force trauma. The presentation of signs and symptoms can be helpful in assessing the decedent; however, in the practice of forensic pathology such a history is more often lacking. PMID- 15894850 TI - Unexpected perinatal death and sudden infant death syndrome (SIDS): anatomopathologic and legal aspects. AB - This work intends to be a review of the recent histopathological findings elicited by research into sudden and unexpected perinatal death and sudden infant death syndrome (SIDS) that have dictated a novel approach to the inherent problems by pathologists, especially those entrusted with forensic medical authority. The new approach stems from the recent advances made in the understanding of neuro- and/or cardiac-conduction-system diseases present in unexpected perinatal death and SIDS. These demand that an accurate morphologic examination be performed of these structures, which modulate respiratory, cardiovascular, digestive, and arousal activities, in all victims of sudden death. A histopathologic study of an ample register of cases of victims of sudden death, either perinatally or in early infancy, has demonstrated frequent alterations both of the autonomic nervous system (especially hypoplasia of the arcuate nucleus) and of the cardiac conduction system (accessory atrioventricular pathways). The present research provides an in-depth study of the many still controversial aspects underlying perinatal unexpected death and SIDS and is recommended for professionals working in the forensic field, whose greater insight into this problem will allow more complete medicolegal documentation. PMID- 15894851 TI - Forensic genetic analysis of insect gut contents. AB - Entomological evidence is most often used for estimating the postmortem interval, but fly larvae can also be a source of vertebrate DNA. Forensic analysis of DNA recovered from a larva's gut can be used to identify what the larva had been feeding on. During our previous research studies, we used the same DNA extraction for the dual purpose of identifying the insect species and associating a maggot with its last meal. In our experience, we have encountered several situations where this method for associating a maggot with a corpse would have been useful, such as removal of remains from a suspected crime scene, an alternative food source is nearby the scene or the body, and a chain-of-evidence dispute. However, since maggot gut content analysis is a quite brand-new area of study, many of the limitations of the technique have not yet been explored. The results of our most recent research studies suggest that third-instar larvae actively feeding on the corpse can be considered the best source of human DNA, better than postfeeding or starved larvae. In this paper, the state of the art of forensic genetic analysis of maggot gut contents is reviewed. PMID- 15894852 TI - Role of chemical tests and scene investigation in determination of range of fire. AB - Forensic pathologists have historically found several characteristics of the entrance wound invaluable in determining range of fire in gunshot fatalities. Among these characteristics are the pattern and constituents of any deposited material such as soot and/or gun powder residues. We describe a case in which the application of previously described characteristics, in the absence of laboratory testing and examination of the crime scene, would have led to an erroneous conclusion with potentially grave consequences. We suggest that all attempts be made to use available laboratory tests and to perform detailed examination of crime scenes in determining the circumstances surrounding fatal gunshot injuries. PMID- 15894853 TI - Accidental death resulting from acetylene cylinder impact. AB - Acetylene is an inflammable gas commonly used for welding in small-scale industries. We present a case of a 34-year-old male welder who died following injuries sustained from explosion of an acetylene gas-welding cylinder. In this case report, we discuss the circumstances leading to the explosion of the welding cylinder, the autopsy findings, and a brief review of the literature on deaths resulting from blasts of acetylene cylinders. PMID- 15894854 TI - Self-inflicted penetrating head injury in a patient with manic-depressive disorder. AB - A 32-year-old Caucasian male with a history of repeated self-injury drilled a hole in his skull using a power tool and subsequently introduced intracerebrally a binding wire from a sketch pad. An emergency craniotomy was performed around the site of cranial injury, and the foreign body was carefully extracted. The wire was located partially in the subdural space and partially in the right hemisphere of the brain. The patient made an excellent recovery and was referred to a psychiatrist for further treatment. This is a rare case of unusual and complex repetitive self-destructive behavior without apparent suicidal intent. The pertinent literature is reviewed and the surgical and psychiatric implications of such injuries are discussed. PMID- 15894855 TI - A suicide using a homemade carbon monoxide "death machine". AB - Deaths related to carbon monoxide (CO) poisoning are common. Most represent accidents and suicides, and most result from CO production via the incomplete combustion of carbon-containing substances. Suicide via CO toxicity is not uncommon and typically involves the use of motor-vehicle exhaust as a source of CO. Presented herein is a case of suicidal CO poisoning in which the CO was produced via a chemical reaction between formic acid and sulfuric acid within a homemade device. PMID- 15894856 TI - Death by sewer gas: case report of a double fatality and review of the literature. AB - Hydrogen sulfide (H2S), the toxic gas associated with the smell of "rotten eggs," is an important cause of work-related sudden death. The gas is particularly insidious due to the unpredictability of its presence and concentration and its neurotoxicity at relatively low concentrations, causing olfactory nerve paralysis and loss of the warning odor. We report a double fatality involving 2 surveyors working near a man-hole, who fell into the sewer and died due to sudden exposure to hydrogen sulfide gas. Key historical, physical, and toxicologic findings are described. Additionally, we present a discussion of the clinical presentations and differential diagnosis, mechanism of injury, metabolism and toxicology, incidence, and scene and safety concerns in fatal hydrogen sulfide exposures. PMID- 15894857 TI - Distinctive patterned injuries caused by an expandable baton. AB - Identification and documentation of patterned blunt-force injuries at autopsy is of utmost forensic importance, particularly when the object or surface producing the injury is unknown or uncertain. Documentation of patterned injuries produced by known objects contributes to the catalogue of forensic knowledge regarding those objects and the injuries they cause. This report presents a case in which a 27-year-old male sustained multiple nonlethal patterned blunt-force injuries produced by an expandable baton and subsequent multiple gunshot wounds during apprehension by police. PMID- 15894858 TI - Pseudolaminar necrosis in cyanide intoxication: a neuropathology case report. AB - We describe the gross and microscopic neuropathological changes in the brain of a 17-year-old male who died 4 days after being poisoned with cyanide. Previous reports indicate that following cyanide intoxication, the brain develops diffuse hypoxic/ischemic changes, predominantly of the basal ganglia. The case we describe here had similar features but in addition showed striking laminar necrosis of the cerebral cortex. This finding in cyanide poisoning has been previously demonstrated by neuroimaging, but not pathologically. PMID- 15894859 TI - West Nile virus encephalitis. AB - West Nile virus (WNV) is a mosquito-borne virus that has caused a large number of deaths in the United States since the first outbreak in New York City in 1998. The outbreak initially was limited to the northeast but has since spread across the entire continental United States. WNV causes a variety of clinical symptoms, but the most severe consequences result from central nervous system infection, resulting in meningitis, encephalitis, or meningoencephalitis. We present a case of a 62-year-old male with metastatic cancer, who died as a result of WNV encephalitis. This is followed by a discussion on the epidemiology of WNV and a detailed summary of the methods and resources available to make a diagnosis of WNV infection postmortem. The material presented in the discussion should provide the forensic pathologist with all the information necessary to make a diagnosis of WNV infection postmortem. If nothing else, the routine collection and storage of serum, cerebrospinal fluid, and tissue for every case can enable the forensic pathologist to make this diagnosis even in cases in which WNV is not suspected until after autopsy. PMID- 15894860 TI - Spontaneous rupture of coronary artery in human immunodeficiency virus-positive patient treated with highly active anti-retroviral therapy (HAART). PMID- 15894861 TI - Decapitation in helmeted motorcyclists. PMID- 15894862 TI - Functional valvular incompetence in decompensated heart failure: noninvasive monitoring and response to medical management. AB - OBJECTIVE: We hypothesized that functional mitral and tricuspid valvular incompetence (MR and TR, respectively) are reversible causes of reduced cardiac output in decompensated heart failure (DF) that accompanies systolic dysfunction in ischemic or nonischemic cardiomyopathy. BACKGROUND: DF, defined as signs and symptoms of heart failure at rest, is rooted in a salt-avid state transduced by neurohormonal activation secondary to impaired renal perfusion. Functional MR and TR are reversible causes of reduced systemic blood flow. Their impact on cardiac output, thoracic fluid content, cardiac chamber dimensions, and valvular apparatus function can be monitored noninvasively, before and after optimized medical management. METHODS: Fourteen male subjects (66 +/- 8 years old) with reduced ejection fraction (24 +/- 5%) secondary to ischemic (71%) or nonischemic (29%) cardiomyopathy, who developed DF with clinical evidence of mitral (MR) and tricuspid (TR) valvular incompetence, were each assessed by bioimpedance and echocardiography before and 1 week after optimized medical management restored compensated failure. RESULTS: Pharmacologic elimination of DF was accompanied by a reduction in body weight (P < 0.01). Hemodynamic improvements included a rise in cardiac index (2.1 to 2.6 L/min/m2; P < 0.01) and a reduction in predicted pulmonary artery systolic pressure (58 to 35 mm Hg; P < 0.001), thoracic fluid content (39 to 32 kOhm; P < 0.001), and systemic vascular resistance (1633 to 1209 dynes/sec/cm5; P < 0.001). Improvements in functional MR and TR included reductions in left and right atrial areas (27 to 24 cm and 26 to 23 cm2, respectively; P < 0.001), color-flow grading of MR and TR severity (P < 0.01), mitral regurgitant volume (105 to 65 mL; P < 0.001), and effective MR orifice size (0.8 to 0.6 cm2; P < 0.01). CONCLUSIONS: In DF, functional MR and TR contribute to reduced cardiac output, increased thoracic fluid content, and systemic vascular resistance, together with enlarged atria and valvular orifice size, which can be improved by medical management. Bioimpedance and echocardiography provide for serial noninvasive assessments of hemodynamic status and valvular function in such cases. PMID- 15894863 TI - Angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers effectively and directly potentiate superoxide scavenging by polymorphonuclear leukocytes from patients with type 2 diabetes mellitus. AB - BACKGROUND: Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin II type 1 receptor blockers (ARBs) have potent antioxidant effects in addition to antihypertensive effects. METHODS: We investigated the ability of ACEIs and ARBs to enhance the superoxide scavenging ability of polymorphonuclear leukocytes (PMNLs) from type 2 diabetic patients (n = 32) and healthy subjects (n = 32). The scavenging ability (U/10(3) cells) of superoxide was measured by electron spin resonance. We used ascorbic acid as a positive control antioxidant and tested captopril, temocapril (an inactive form of ACEI), and temocaprilate (an active form of ACEI) as ACEIs, as well as RNH-6270 as an ARB. RESULTS: Captopril, temocaprilate, and RNH-6270 showed dose-dependent enhancement in scavenging ability. The scavenging ability with captopril and temocaprilate was greater than with RNH-6270. The changes in scavenging ability induced by all of the drugs in diabetic patients were similar to the changes in healthy subjects. A high-glucose medium (400-800 mg/dL) greatly attenuated the drug-induced enhancement of scavenging ability. CONCLUSIONS: We demonstrated that both ACEIs and ARBs enhance superoxide scavenging by PMNLs from type 2 diabetic patients and that a high glucose environment markedly attenuates the ability of these drugs to augment superoxide scavenging. PMID- 15894864 TI - Similar decline in post-myocardial infarction mortality among subjects with and without diabetes. AB - BACKGROUND: Data from the 1970s and 1980s suggest that the rate of mortality from coronary disease for patients with diabetes has changed less than that for patients without diabetes. We evaluated trends in post-myocardial infarction mortality and morbidity in patients with and without diabetes over a 7-year period from 1990 through 1997, when substantial changes occurred in the management of coronary disease. METHODS: All patients discharged with the primary diagnosis of acute myocardial infarction (MI) from any Veterans Affairs Medical Center in the country between October 1990 and September 1997 were identified. Demographic, comorbid conditions, inpatient, outpatient, mortality, and readmission data were extracted. Mortality, trends in mortality over time, revascularization, readmissions, and length of hospital stay for MI were compared for the group with diabetes and the group without diabetes. Independent predictors of survival using a Cox regression model were examined. RESULTS: We identified 67,889 patients with MI, of whom 17,756 (26%) had diabetes. At 60 days post-MI, there was a 29% higher mortality rate in the group with diabetes (5.2% versus 4.0%, P < 0.001), which increased to 35% at 1 year (16.1% versus 11.9%, P < 0.001). Diabetes was independently associated with increased overall mortality. Age-adjusted 1-year post-MI mortality from 1991 to 1998 had a significant downward trend (4.9% decrease in odds of mortality per year, P < 0.001) regardless of diabetes status. CONCLUSIONS: Patients with diabetes showed a trend toward declining 1-year post-MI mortality rate that was not significantly different from that seen in patients without diabetes. Further work needs to be done to narrow the gap between the two groups. PMID- 15894865 TI - Nontyphoid Salmonella bacteremia in patients with liver cirrhosis. AB - BACKGROUND: Bacteremia is reported to occur in 4% to 9% of hospitalized patients with liver cirrhosis. Escherichia coli and Klebsiella pneumoniae are the most commonly isolated organisms. Only sporadic cases of nontyphoid Salmonella bacteremia are reported in the literature. In this study, we sought to determine the clinical features and prognosis of patients with liver cirrhosis and bacteremia due to nontyphoid Salmonella. METHOD: Data were collected by retrospective chart review. RESULTS: From December 1996 to May 2003, we identified 23 patients (18 males, 5 females) with a median age of 58 years. The Child classification for liver cirrhosis was A in 4, B in 9, and C in 10 patients. Solid organ cancers were present in 14 patients: hepatocellular carcinoma in 13 patients and gastric carcinoma in 1 patient. Hospital death occurred in 11 patients (48%): 7 died of sepsis and 4 of hepatic failure. Using a logistic regression model, the independent risk factors for death in patients with nontyphoid Salmonella bacteremia were young age and an advanced stage of liver cirrhosis. CONCLUSION: Most nontyphoid Salmonella bacteremia in patients with liver cirrhosis was community-acquired. An advanced stage of liver cirrhosis and hepatocellular carcinoma were common. The prognosis for young patients was unfavorable; this was seemingly due to hepatocellular carcinoma producing more unfavorable results in younger cirrhotic patients. PMID- 15894866 TI - The use of implantable cardioverter defibrillators for the prevention of sudden cardiac death: a review of the evidence and implications. AB - Sudden cardiac death (SCD) claims approximately 460,000 lives per year in the United States, and half of these deaths occur in people with a history of coronary artery disease. Patients with left ventricular dysfunction and a history of myocardial infarction are at especially high risk. There is now strong evidence from multiple well-designed randomized controlled trials that implantable cardioverter defibrillators (ICDs) save lives when used for both primary and secondary prevention. As indications for ICD implantation have broadened, considerable debate has taken place because of the substantial cost involved in widespread ICD utilization. This article summarizes the epidemiology of SCD, reviews the evidence supporting the use of ICDs in patients with ischemic cardiomyopathy, and explores some of the controversy surrounding ICD utilization that has arisen in the wake of recent trials that have utilized ICDs for the primary prevention of SCD. PMID- 15894867 TI - The emerging role of endoscopic ultrasonography in cancer staging. AB - The management of solid organ cancers relies on accurate staging. Once distal metastasis has been excluded by conventional radiographic methods, local staging of the tumor and its nodal involvement is essential to determine the most optimal therapeutic approach. Many radiographic imaging modalities are unable to determine subtle tumor involvement. However, the emergence of endoscopic ultrasonography (EUS) has given promise to improved staging with its unique ability to examine tumors from within the gastrointestinal lumen with extremely close proximity. An additional advantage is the ability to perform fine-needle aspiration (FNA) biopsy to confirm or exclude tumor involvement. Many studies have shown superior accuracy in staging with EUS and EUS-FNA for tumors of the esophagus, stomach, pancreas, rectum, and mediastinum, including lung cancer. This review illustrates the principles of EUS and its role in staging of a variety of cancers and particularly its role alongside other imaging modalities. PMID- 15894868 TI - Hypoglycemia and hyperglycemia due to insulin antibodies against therapeutic human insulin: treatment with double filtration plasmapheresis and prednisolone. AB - Two diabetic patients, who had been treated with human insulin, suffered from fasting hypoglycemia and postprandial hyperglycemia. The insulin-binding capacities of their sera were high, and a large amount of total insulin and prolonged presence of free insulin in the sera were shown. Scatchard analysis of these insulin antibodies revealed that high-affinity insulin antibodies had larger capacity and stronger affinity compared with commonly insulin-treated patients. Treatment with double filtration plasmapheresis and subsequent administration of prednisolone in the second patient reduced such antibodies and resulted in recovery of glycemic control by insulin. Hypoglycemia and hyperglycemia could be incurred when insulin antibodies with strong affinity and high capacity in high-affinity sites arise. This condition can be treated with double filtration plasmapheresis and subsequent administration of prednisolone. PMID- 15894869 TI - Vascular involvement in a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes. AB - A 26-year-old man with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) was admitted to our hospital for further cardiovascular examination. A muscle biopsy demonstrated strongly succinate dehydrogenase-reactive blood vessels. Pulse wave contour analysis revealed that both capacitive and oscillatory compliance were markedly reduced in this patient compared with 45 normal age-matched control subjects. Hepatocyte growth factor was remarkably elevated in this patient over that of 10 normal control subjects. These findings suggest that a MELAS patient has not only pathologic but also functional vascular involvement. If so, patients with MELAS need systemic vascular assessment. PMID- 15894870 TI - Successfully treated mitral valve Proteus mirabilis endocarditis. AB - Infective endocarditis caused by Proteus mirabilis is rare and is fatal in most cases. A case of a 64-year-old man presenting with fever, dysuria, and murmur is reported. P mirabilis endocarditis was diagnosed based on clinical presentation, blood culture findings, and the presence of a large mitral valve vegetation seen on echocardiogram. The patient survived after long-term antibiotics and surgical replacement of the mitral valve. The clinical characteristics and complications from reported cases of P mirabilis endocarditis are reviewed. PMID- 15894872 TI - Advances and methods in liver surgery: haemostasis. PMID- 15894873 TI - UV Overdose vs Hyperoxia. PMID- 15894874 TI - The diagnosis and characteristics of moderate dry eye in non-contact lens wearers. AB - PURPOSE: To identify and characterize moderate dry eye in non-contact lens wearers with a new scoring system-based dry eye questionnaire and to determine which objective tests better differentiate patients with moderate dry eye from healthy patients. METHODS: Fifty-two healthy subjects (21 women and 31 men with a mean age of 27.8 +/- 9.2 years) and 37 subjects with moderate dry eye (33 women and 4 men with a mean age of 36.4 +/- 12.9 years) completed a 42-item dry eye questionnaire. Seventeen healthy subjects (11 women and 6 men with a mean age of 30.5 +/- 9.7 years) and 28 subjects with moderate dry eye (24 women and 4 men with a mean age of 38.50 +/- 3.8 years) underwent additional objective assessment of ocular surface health, tear osmolality, tear stability, and tear volume. RESULTS: Subjects with moderate dry eye scored significantly higher (49.8 +/- 20.3, P<0.0001) on the dry eye questionnaire than did normal subjects (11.7 +/- 10.3). Ocular irritation symptoms worsened with progression of time of day in both groups of subjects. Internal reliability (0.95 Cronbach alpha) was excellent, and concurrent validity (Spearman rho 0.507) was acceptable when compared to the McMonnies and Ho dry eye questionnaire. Significant differences in tear osmolality (P<0.00001), invasive tear breakup time (P<0.034), and corneal vital dye staining (P<0.0001) were detected between the two groups of subjects. A stepwise linear regression on objective clinical tests, however, did not account for 77% of the total variance in the questionnaire scores. CONCLUSIONS: A unique scoring system-based dry eye questionnaire was validated to separate non-contact lens wearers with moderate dry eye from healthy subjects. Objective tests of tear osmolality and stability and ocular surface integrity were better than other clinical measures at identifying differences between the two subject groups. The results strongly support the evidence that the diagnosis and treatment of moderate dry eye requires a detailed assessment of self-perceived symptoms and that objective clinical testing alone may be insufficient. PMID- 15894875 TI - Safety and efficacy of the technique of complete tenon's membrane excision and mitomycin C in pterygium surgery. AB - PURPOSE: Despite the established efficacy of mitomycin C (MMC) in preventing recurrence after pterygium surgery, there is no consensus on the optimal operative technique or dose of MMC. METHODS: The authors retrospectively evaluated 90 single-headed pterygium resections performed with excision of the entire pterygium to the plica and removal of all Tenon's membrane from the scleral bed and rectus muscle. Bare sclera was exposed to 0.4 mg/mL MMC for 5 minutes for primary pterygia and 6 minutes for recurrent pterygia. These procedures were compared to a previous series of 58 procedures using bare sclera technique with MMC, but without extensive Tenon's membrane removal. RESULTS: Extensive Tenon's membrane removal was associated with significantly fewer recurrences (6.7% vs. 21%; P< or =0.025) and significantly less pyogenic granuloma formation (24.4% vs. 40%; P< or =0.05). In addition, there were no vision-threatening complications, such as scleral melting or persistent corneal epithelial defects, in any of the patients. CONCLUSIONS: These findings suggest that complete removal of all Tenon's membrane in the affected quadrant combined with topical MMC in the aforementioned dose is a safe method of pterygium excision with a low rate of recurrence and pyogenic granuloma formation. PMID- 15894876 TI - Ocular surface injury from a microwave superheated egg resulting in a pseudopterygium. AB - PURPOSE: To describe the first case of ocular surface injury resulting in a pseudopterygium from a microwave superheated egg. METHODS: Case report. RESULTS: A 12-year-old girl sustained an ocular surface injury resulting in a pseudopterygium from a microwave superheated egg. CONCLUSION: Microwave superheated eggs can result in ocular injury. This case illustrates the potential ocular danger involved with microwave ovens. PMID- 15894877 TI - Conductive keratoplasty and the coupling phenomenon. AB - PURPOSE: To determine whether changes in corneal astigmatism with astigmatic conductive keratoplasty (CK) treatment obey Gaussian coupling (i.e., the steepening of the flat axis associated with the flattening of the steep axis) and to measure the coupling ratio and the coupling constant to determine the effect of astigmatic CK treatment on spherical equivalent. METHODS: Retrospective review of 33 eyes in 24 patients who had undergone CK for hyperopia and who were treated intraoperatively for induced astigmatism. Induced astigmatism was determined by comparison of keratometric readings before and after CK. RESULTS: The coupling ratio was calculated according to two equations: clinical coupling ratio (1.61 +/ 0.81 diopters) and coupling ratio of the surgically induced refractive change (1.57 +/- 1.16 diopters). Values of the coupling ratio between 0.72 and 1.88 indicate coupling. CONCLUSIONS: The cornea does not behave according to Gauss's law of elastic domes. The targeted flat axis and steep axis 90 degrees away are affected by CK in opposite but not equal amounts. Therefore, when correcting surgically induced astigmatism with CK, the overall spherical equivalent of the patient will change because the coupling ratio is not equal to 1. PMID- 15894878 TI - Ocular surface temperature: a review. AB - PURPOSE: To review the evolution in ocular temperature measurement during the last century and examine the advantages and applications of the latest noncontact techniques. The characteristics and source of ocular surface temperature are also discussed. METHODS: The literature was reviewed with regard to progress in human thermometry techniques, the parallel development in ocular temperature measurement, the current use of infrared imaging, and the applications of ocular thermography. RESULTS: It is widely acknowledged that the ability to measure ocular temperature accurately will increase the understanding of ocular physiology. There is a characteristic thermal profile across the anterior eye, in which the central area appears coolest. Ocular surface temperature is affected by many factors, including inflammation. In thermometry of the human eye, contact techniques have largely been superseded by infrared imaging, providing a noninvasive and potentially more accurate method of temperature measurement. Ocular thermography requires high resolution and frame rate: features found in the latest generation of cameras. Applications have included dry eye, contact lens wear, corneal sensitivity, and refractive surgery. CONCLUSIONS: Interest in the temperature of the eye spans almost 130 years. It has been an area of research largely driven by prevailing technology. Current instrumentation offers the potential to measure ocular surface temperature with more accuracy, resolution, and speed than previously possible. The use of dynamic ocular thermography offers great opportunities for monitoring the temperature of the anterior eye. PMID- 15894879 TI - Microbial changes in conjunctival flora with 30-day continuous-wear silicone hydrogel contact lenses. AB - PURPOSE: To determine the effect of 30-day continuous-wear silicone hydrogel contact lenses on the conjunctival flora in asymptomatic wearers. METHODS: The authors studied 29 eyes of 15 patients wearing Focus NIGHT & DAY silicone hydrogel contact lenses for up to 30 nights of continuous wear. The average age of the patients was 25.54 +/- 8.98 years. Cultures of the inferior cul-de-sac were taken bilaterally from all eyes, before and after lens wear in asymptomatic patients. The isolation and identification of bacteria were made by standard clinical laboratory methods. RESULTS: The number of eyes whose conjunctival cultures were sterile before using the lenses significantly decreased (P = 0.0005), and the number of eyes with a growth of coagulase-negative staphylococci and diphtheroid rods in their conjunctival cultures significantly increased after using these lenses (P = 0.001 and P = 0.031, respectively). Conversely, a statistically significant difference was not found in the number of eyes that carried Propionibacterium acnes and Fusobacterium nucleatum in their conjunctival cultures before and after using the 30-day continuous-wear silicone hydrogel lenses (P = 0.998 and P = 0.488, respectively). CONCLUSIONS: The results suggest that the sterility of the conjunctiva significantly decreased after using 30-day continuous-wear silicone hydrogel contact lenses. In addition, the number of bacteria of the normal conjunctival flora significantly increased after the use of these lenses. Contamination by the bacteria of the eyelids may be a possible colonization factor in this study group. Therefore, it is appropriate to examine the patients who wear these lenses more frequently. PMID- 15894880 TI - Reverse-geometry gas-permeable lens design for pellucid marginal degeneration. AB - PURPOSE: To report a case of treating a patient with pellucid marginal degeneration (PMD) by using a reverse-geometry gas-permeable lens design. METHODS: A 38-year-old Hispanic man was referred for having reduced visual acuity secondary to distorted corneas. The patient was diagnosed with PMD after an extensive slitlamp examination showed a thinning of the inferior peripheral cornea in both eyes. Corneal topography (Orbscan II) was performed to help confirm the diagnosis. A Reverse Aspheric Ortho Focus (RAOF) gas-permeable lens was fitted on this patient. RESULTS: The Orbscan II corneal topography showed distinct peripheral steepening, a pronounced astigmatic pattern (greater in the left eye than in the right), and thinner corneas in the inferior periphery than centrally. The fluorescein pattern of the RAOF 5 lenses showed central alignment, good edge lift 360 degrees, mid peripheral bearing with 0.5 mm vertical movement on blinking, and good centration. The patient's distance visual acuity with the lenses was 20/20-1 in the right eye and 20/20 in the left. CONCLUSIONS: The Orbscan II was an important tool in making the diagnosis of early PMD, because few clinical signs were observed. Not only does the Orbscan II provide the clinician with a topographic map of the cornea, but it also measures the corneal thickness, which aided in the diagnosis of this patient. In addition, the posterior float measurement provided by the Orbscan II may be instrumental in making a differential diagnosis. A reverse-geometry gas-permeable lens provided the patient with improved peripheral fit over conventional designs, adequate comfort, and optimal visual acuity. PMID- 15894881 TI - Fluid-ventilated, gas-permeable scleral contact lens is an effective option for managing severe ocular surface disease and many corneal disorders that would otherwise require penetrating keratoplasty. AB - PURPOSE: To examine the diagnostic indications and relative merits of a fluid ventilated, gas-permeable scleral lens for improving vision impaired by irregular astigmatism and for providing a therapeutic environment for managing severe ocular surface disease. METHODS: After a review of scleral lens development and a description of current design and manufacturing innovations, indications for fitting the Boston Scleral Lens were evaluated based on a retrospective review of all available records of patients fitted with this device, and outcome experiences were described. RESULTS: A total of 875 eyes of 538 patients were fitted with the fluid-ventilated, gas-permeable scleral lens during the past 18 years. Most patients were fitted in the past 4 years. Indications included managing severe ocular surface disease and rehabilitating vision impaired by irregular astigmatism associated with corneal disorders. Rigid gas-permeable contact lenses either were not tolerated or were contraindicated in all eyes. Outcomes included improved vision and reduced ocular pain and photophobia associated with severe ocular surface disease. Scleral lenses promoted healing of persistent epithelial defect (PED) refractory to other treatments and prevented PED recurrence in stem cell-deficient and neurotrophic corneas. Microbial keratitis occurred in 4 of 22 eyes treated with extended scleral lens wear for PED after penetrating keratoplasty. CONCLUSIONS: The fluid-ventilated, gas permeable scleral lens is an important front-line tool for managing many corneal disorders refractory to other treatment measures or otherwise requiring keratoplasty. PMID- 15894883 TI - Evidence-based medicine in hypertension: what type of evidence? AB - Although placebo-controlled trials have been a powerful instrument to answer the question whether blood pressure lowering in hypertensive patients were beneficial or not, event-based trials comparing different active treatments have been less successful, and have generated the opinion that blood pressure lowering is the only relevant beneficial mechanism. Several limitations of these event-based trials should be considered, however only a proportion of the patients have received the initial randomized treatment alone, the goal reducing blood pressure level has rarely been achieved and small blood pressure differences have often been underestimated, patients have been randomized to given regimens as if blood pressure of all them should be equally responsive to different agents, and, finally, most trials have only included elderly or complicated hypertensive. The hypothesis is advanced that blood pressure lowering and ancillary properties of antihypertensive agents may play a different role at various stages of the continuum of cardiovascular disease. Only when hypertension-related complications are not too advanced, the different ability of different antihypertensive agents to influence subclinical organ damage progression ("intermediate" endpoints) may be capable of influencing morbidity and mortality over a longer term than that usually explored by event-based trials. PMID- 15894884 TI - Preventing pregnancy-induced hypertension: are there regional differences for this global problem? AB - Developing countries have had persistently higher rates of maternal and child mortality due to pre-eclampsia in comparison with developed countries. Moreover, evidence from studies of interventions to prevent pre-eclampsia have given contradictory results. In this review, we discuss the underlying causes of pre eclampsia, and the results of clinical trials performed to prevent this disease, that support the proposal that the causes and strategies to prevent pre-eclampsia are different in developed and developing countries. We also suggest that the establishment of an adequate prenatal care is the only effective way to reduce the incidence of pre-eclampsia in populations from developing countries, especially in women at high risk of pregnancy-induced hypertension. PMID- 15894885 TI - Nocturnal fall of blood pressure with antihypertensive therapy and recurrence of ischaemic stroke: 'the lower the better' revisited. PMID- 15894886 TI - Mesenchymal stem cells: unknown mechanisms of differentiation. PMID- 15894887 TI - How to manage blood pressure in acute stroke. PMID- 15894888 TI - Assessment of 24-h neuroadrenergic profile: does the dream come true? PMID- 15894889 TI - Interarm differences in seated systolic and diastolic blood pressure: the Hypertension Genetic Epidemiology Network study. AB - OBJECTIVE: To examine whether blood pressure (BP) differs between arms in hypertensive siblings and randomly selected volunteers, and whether this difference is explained by cardiovascular risk factors. METHODS: The Hypertension Genetic Epidemiology Network recruited 2395 hypertensive siblings and 854 volunteers. BP was measured six times (three measurements per arm) in seated participants using a Dinamap monitor. The average of three measurements was calculated per arm and the difference taken between arms (i.e. interarm BP differences). RESULTS: The mean age of the subjects was 56 years, and about one half of the sample was male. More than one-half of the sample was African American. The mean diastolic BP was equal in the two arms in the random sample (68.8 versus 68.7 mmHg) and in hypertensive siblings (73.4 versus 73.1 mmHg), as was systolic BP (random, 119.6 versus 119.3 mmHg; hypertensives, 130.8 versus 130.7 mmHg). The mean interarm diastolic and systolic BP differences were 2.96 +/ 2.51 and 4.61 +/- 4.10 mmHg, respectively, in the random sample and were 3.09 +/ 2.73 and 5.35 +/- 4.98 mmHg, respectively, in hypertensive siblings. Few (random, 1.6%; hypertensives, 2.8%) had interarm diastolic BP differences > 10 mmHg, but 9.2% of the random sample and 14.2% of hypertensive siblings had systolic BP differences > 10 mmHg. Obesity, higher heart rate, and higher systolic BP were associated with larger interarm BP differences. These results have implications for blood pressure measurement in research settings and in screening programs. PMID- 15894890 TI - Correlation between left ventricular mass and urinary sodium excretion in specific genotypes of CYP11B2. AB - BACKGROUND: Aldosterone has essential roles in regulating intravascular volume and blood pressure, and is suggested to influence cardiac structure. However, the association of polymorphisms in the aldosterone synthase gene (CYP11B2) with hypertension or cardiac hypertrophy remains controversial. OBJECTIVE: To evaluate the distribution of polymorphisms in the CYP11B2 gene and the possible associations between genotypes and blood pressure, urinary excretion of aldosterone or electrolytes and echocardiographic measurements, in a Japanese population. METHODS AND RESULTS: We examined the association of two common diallelic polymorphisms within CYP11B2, one in the promoter -344T/C and the other an intron 2 gene conversion, with blood pressure, 24-h urinary excretion of aldosterone and electrolytes, and echocardiographic measurements, in a Japanese population. We confirmed significant linkage disequilibrium between these polymorphic loci and ethnic differences in frequency of the alleles. The -344C and -344T haplotypes apparently diverged before the intron conversion polymorphism was generated on the latter haplotype. Allele frequencies did not differ between 535 normotensive and 360 hypertensive individuals or between hypertensive individuals with higher and lower concentrations of renin. The only significant correlation was a positive correlation of left ventricular mass with 24-h urinary excretion of sodium, which occurred only in individuals with the 344CC genotype or the intron 2 conversion (-/-) genotype. CONCLUSIONS: The -344CC or intron 2 conversion (-/-) genotype in CYP11B2 may be a risk factor for developing sodium-sensitive cardiac hypertrophy. Ethnic differences in the distribution of CYP11B2 genotypes combined with differences in salt intake might account for inconsistencies between previous reports. PMID- 15894891 TI - Aldosterone synthase gene polymorphism, stroke volume and age-related changes in aortic pulse wave velocity in subjects with hypertension. AB - PURPOSE: In rats, chronic aldosterone administration with high diet intake increases aortic stiffness independent of mechanical stress. In hypertensive humans, enhanced plasma aldosterone and arterial stiffness are positively associated. Whether the aldosterone synthase gene polymorphism (ASGP) CYP11B2 influences the age-related changes in blood pressure (BP) and arterial stiffness in hypertensive subjects has never been investigated. METHODS: In 425 untreated hypertensive men and women, ASGP was evaluated together with aortic pulse wave velocity (PWV). In 191 of these subjects, cardiac haemodynamics were measured using echo-Doppler techniques. RESULTS: In the overall population, independently of sex, the TC and CC genotypes of ASPG had significantly higher heart rate (HR) (P < 0.05) and lower stroke index (P < 0.01) than the TT genotype, but did not affect BP. In men, the adjusted slopes of the curves relating age to PWV and HR were significantly steeper (P = 0.04; P = 0.002) for the TC and CC than for the TT genotype. Such gene-related differences were not observed for the age-systolic BP relationship. CONCLUSION: In hypertensive subjects, the TC and CC genotypes of ASGP involve, by comparison with the TT genotype, significantly higher HR and reduced stroke index. In men with the C allele, the reduced stroke index (cardiac effect) compensates for the steep increase of PWV with age (arterial effect), thus modulating the cardiovascular phenotype and explaining the lack of increased incidence of systolic hypertension. The results are consistent with a local role of endogenous aldosterone on both heart and vessels. PMID- 15894892 TI - Participation of aldosterone in the vascular inflammatory response of spontaneously hypertensive rats: role of the NFkappaB/IkappaB system. AB - OBJECTIVE: To investigate the participation of aldosterone in the vascular inflammatory process associated with hypertension, as well as the possible involvement of the NFkappaB/IkappaB system. METHODS: Male spontaneously hypertensive rats (SHR; 20-22 weeks old) untreated or treated with either the aldosterone receptor antagonist, eplerenone (100 mg/kg per day) or triple antihypertensive therapy (HHR: hydralazine + hydrochlorothiazide + reserpine; 20 + 7 + 0.15 mg/kg per day) were used in the study. Wistar-Kyoto rats (WKY) were used as a normotensive reference group. Aortic mRNA expression and plasma levels of interleukin (IL)-1beta, IL-6 and tumour necrosis factor alpha (TNFalpha) were measured. Likewise, the aortic expression of the nuclear factor kappaB (NFkappaB) p50 subunit precursor, p105, and its inhibitor (IkappaB) were measured. RESULTS: SHR showed higher aortic expression of IL-1beta, IL-6 and TNFalpha than WKY (P < 0.05) and higher plasma levels of IL-1beta and IL-6 than WKY (P < 0.05). Moreover, SHR also presented increased aortic expression of nuclear transcription factor NFkappaB p50 subunit precursor (p105), and a reduction of its inhibitor IkappaB. Both eplerenone and HHR decreased blood pressure to a comparable extent (P < 0.05). This effect was accompanied by a reduction in plasma levels of IL 1beta and IL-6 and aortic mRNA expression of IL-1beta, IL-6 and TNFalpha. However, the effect of eplerenone was more marked, since eplerenone-treated rats showed significantly lower inflammatory parameters than SHR receiving HHR. In addition, both antihypertensive treatments increased IkappaB mRNA expression in a similar manner, but only eplerenone reduced NFkappaB mRNA expression. CONCLUSIONS: Aldosterone, as well as an increase in haemodynamic forces produced by hypertension, participate in the vascular inflammatory process associated with hypertension in SHR. This effect seems to be mediated by enhanced vascular expression of cytokines through a modification of the NFkappaB/IkappaB system. PMID- 15894893 TI - Chronic reductions in carotid blood flow cause salt-sensitive hypertension in rats. AB - OBJECTIVE: We determined whether chronic reductions in carotid blood flow elicit salt-sensitive hypertension through regulation of the brain renin-angiotensin system (RAS). DESIGN AND METHODS: Both internal carotid arteries of male Wistar rats were ligated over a 1-week period. Carotid-ligated or sham-operated rats were treated with a high-salt (8% NaCl diet and 1% NaCl drinking water) or low salt (0.3% NaCl diet and distilled water) diet for 6 weeks. At the end of the experiment, expression of the RAS mRNAs in the brain was measured. Effects of a 6 day intracerebroventricular infusion of CV-11974, a selective non-peptide angiotensin II type 1 (AT1) receptor blocker, were investigated in carotid ligated rats administered high-salt diet. RESULTS: High-salt administration increased systolic arterial pressure compared with low-salt administration in sham-operated rats [168 +/- 4 mmHg (n = 10) versus 149 +/- 3 mmHg (n = 10), P < 0.001] and in carotid-ligated rats [202 +/- 5 mmHg (n = 10) versus 153 +/- 2 mmHg (n = 10), P < 0.0001]. Systolic arterial pressure, urinary excretion of vasopressin and norepinephrine, and expression of renin, angiotensin I converting enzyme, and AT1 receptor mRNAs in the hypothalamus were greater in carotid ligated rats than in sham-operated rats treated with high salt. In contrast, these parameters did not differ between carotid-ligated and sham-operated rats treated with low salt. Intracerebroventricular infusion of CV-11974 abolished the increase in these parameters in carotid-ligated rats treated with the high-salt diet. CONCLUSIONS: Chronic reductions in carotid blood flow may cause salt sensitive hypertension in normotensive rats by activating the hypothalamic RAS. PMID- 15894894 TI - Stimulation of NADPH oxidase by angiotensin II in human neutrophils is mediated by ERK, p38 MAP-kinase and cytosolic phospholipase A2. AB - OBJECTIVE: The present research was designed to study the involvement of ERK and p38 MAP-kinase in cytosolic phospholipase A2 (cPLA2) and NADPH-oxidase activation by angiotensin II (Ang II) in human neutrophils. METHODS: NADPH-oxidase activity was measured by reduction of cytochrome C. cPLA2 activity was measured in cell lysate using sonicated dispersions of 1-stearoyl-2-[C]arachidonyl phosphatidylcholine. Cells were incubated with MEK inhibitor UO126 or with p38 MAP-kinase inhibitor SB202190 prior to stimulation with Ang II. Translocation of p47, p67 and cPLA2 and phosphorylation of ERK and p38 MAP-kinase were measured by immunoblot analysis. RESULTS: Ang II induced a dose-dependent activation of NADPH oxidase in neutrophils and monocytes as well as in differentiated PLB-985 cells towards neutrophil or monocyte lineages, but not in cPLA2-deficient differentiated PLB-985 cells. An immediate activation of both ERK and p38 MAP kinase and of cPLA2 was induced by Ang II in human neutrophils. In addition, Ang II induced translocation of the cytosolic oxidase components, detected by translocation of p47, which preceded the translocation of cPLA2 induced by this agonist. The p38 MAP-kinase inhibitor SB202190 or the MEK-ERK pathway inhibitor UO126 totally inhibited the activation of both NADPH oxidase and cPLA2 as well as the translocation of cytosolic oxidase components and of cPLA2 to the membrane fractions. CONCLUSIONS: These results suggest that either ERK or p38 MAP-kinase are involved in the activation of both cPLA2 and NADPH oxidase, and that cPLA2 is required for activation of the NADPH oxidase by Ang II in human neutrophils. PMID- 15894895 TI - Differential regulation of smooth muscle markers in human bone marrow-derived mesenchymal stem cells. AB - OBJECTIVE: To study smooth-muscle differentiation and de-differentiation of human bone marrow-derived mesenchymal stem cells (MSCs), which have been shown to enter the circulation and to contribute to vascular repair and atherosclerosis. DESIGN: Human MSCs from bone marrow were cultured with 20% fetal calf serum (FCS) or with 10% FCS and various concentrations of dimethyl sulfoxide (DMSO). Expression of smooth muscle markers was determined by Western blot analysis and immunofluorescence. For signalling studies, involvement of the mammalian target of rapamycin (mTOR) pathway was tested by treatment with rapamycin. RESULTS: MSCs cultured with 20% FCS acquired a smooth muscle-like appearance and expressed the smooth muscle (sm) markers sm-alpha-actin, desmin, sm-calponin and myosin light chain kinase (MLCK). DMSO induced a spindle-like morphology with marked reduction of stress fibers. As judged by Western blot analysis, treatment with 2.5% DMSO strongly downregulated expression of sm-calponin (-85%), short MLCK (-98%) and sm alpha-actin expression (-51%). Reduced calponin expression was detected by day 2 of treatment with 0.5-2.5% DMSO. After withdrawal of DMSO, MSCs regained high expression of sm-calponin. Treatment with 6 nmol/l rapamycin partly antagonized the effect of DMSO, indicating the involvement of mTOR in regulation of the smooth muscle phenotype of MSCs. CONCLUSIONS: DMSO strongly downregulates the smooth muscle markers sm-calponin, short MLCK and sm-alpha-actin in human MSCs, indicating a transition from a smooth muscle-like phenotype to an undifferentiated state by an mTOR-dependent mechanism. Regulating the phenotype of human MSCs may be of relevance for novel therapeutic approaches in atherosclerosis and intimal hyperplasia after vascular injury. PMID- 15894896 TI - Carotid wall composition in hypertensive patients after 4-year treatment with lacidipine or atenolol: an echoreflectivity study. AB - OBJECTIVE: In the European Lacidipine Study on Atherosclerosis (ELSA), against a similar antihypertensive effect, a significantly greater effect of lacidipine was found on carotid intima-media thickness (IMT) progression, indicating a specific anti-atherosclerotic effect of lacidipine. However, not only the extent but also the composition of an atherosclerotic plaque is a determinant of subsequent events. DESIGN AND METHODS: Among the 2334 patients enrolled in ELSA, 420 with 4 year treatment were chosen, videodensitometric analysis of their ultrasound carotid scan was performed and the maximum wall lesion (Tmax) was classified as lipidic, fibrolipidic and fibrotic by means of software previously validated against histology. Of the 420 patients, 244 had scans suitable for videodensitometry. RESULTS: Excellent reproducibility of videodensitometry analysis was found using the Bland-Altman and other methods. At baseline, ELSA hypertensive patients had predominantly fibrolipidic walls (70%), with an echoreflectivity indicating a mean collagen content of 25%. After 4 years of antihypertensive treatment, little change in the frequency distribution of carotid lesions (fibrolipidic 73%) was found, with no significant differences between patients randomized to lacidipine or atenolol. CONCLUSIONS: Our study provides previously unavailable information that carotid wall composition changes to an extremely small extent during 4-year effective blood pressure lowering. With lacidipine, stable composition is associated with a lower IMT progression than with atenolol. PMID- 15894897 TI - Arterial stiffness is an independent risk factor for cognitive impairment in the elderly: a pilot study. AB - BACKGROUND: Loss of cognitive function is a common condition in the elderly population. Cognitive impairment is defined as the transitional stage of cognitive decline, between normal aging and early dementia. We tested whether arterial stiffness, evaluated as pulse wave velocity (PWV), is associated with cognitive impairment in older subjects, and whether PWV is increased at a comparable extent in older subjects with cortical or subcortical cerebral lesions when compared with age-matched controls referred for memory deficits. SUBJECTS AND METHODS: Eighty-four subjects (78 +/- 5 years, 30 men and 54 women) referred for memory deficit with no history of stroke or atrial fibrillation were studied. Carotid-femoral PWV was determined non-invasively with Complior. The Mini Mental State Examination was assessed as a measure of global cognitive function. The sum of the score on the Activities of Daily Living and Instrumental Activities of Daily Living scales was used as a measure of personal independency. Based upon brain imaging, subjects were classified as referred for memory deficits with normal brain imaging, or control, with subcortical microvascular lesions or with cortical atrophy. RESULTS: PWV, normalized for mean blood pressure, was inversely correlated with the Mini Mental State Examination (r = -0.26, P < 0.05), even after controlling for education, prevalent cardiovascular (CV) disease, CV risk factors, and medication use (beta coefficient = -0.28, P < 0.01). PWV was also inversely correlated with personal independency (r = -0.36, P < 0.01; beta coefficient = -0.38, P < 0.01, after multiple adjustment). In the presence of no significant differences in age, education, traditional CV risk factor levels, carotid plaques, or prevalence of CV disease, higher PWV values were more frequent in subjects with cortical atrophy than in patients with subcortical microvascular lesions or controls (P < 0.05). CONCLUSIONS: PWV was associated with cognitive impairment and with a greater personal dependency, independently of major modifiable CV risk factors. PMID- 15894898 TI - Effects of blood pressure levels on case fatality after acute stroke. AB - OBJECTIVE: We evaluated the relationship between admission blood pressure (BP) and early prognosis in patients with acute stroke in a single cohort. DESIGN: The subjects comprised 1004 cases of brain infarction and 1097 cases of brain hemorrhage, who were admitted to hospitals on the day of stroke onset. Death within 30 days after onset was evaluated in relation to admission BP levels. RESULTS: In brain infarction, a U-shaped relationship was found between BP levels and mortality rate, with a nadir at systolic blood pressure (SBP) of 150-169 mmHg and at diastolic blood pressure (DBP) of 100-110 mmHg. After adjustments for age and sex, the highest relative risks (RR) was observed in the lowest BP levels compared with nadir groups, and were 2.69 [95% confidence interval (CI), 1.43 5.07] in SBP and 3.49 (95% CI, 1.58-7.74) in DBP. In subjects with previous hypertension, the relationship between prognosis and SBP level shifted significantly toward higher pressure by about 10 mmHg compared with those without previous hypertension. In subjects with brain hemorrhage, the relationship between BP levels and mortality rate showed a J-shape in SBP and a U-shape in DBP. Highest BP levels had the poorest prognoses (>/= 230 mmHg in SBP, RR = 4.13, 95% CI = 2.45-6.94; >/= 120 mmHg in DBP, RR = 1.83, 95% CI = 1.11-3.04). CONCLUSION: Lower and higher BP after brain infarction and higher BP after brain hemorrhage were predictors for poor early prognosis. In subjects with brain infarction, patients with previous hypertension had better outcomes at higher admission BP level than did normotensive patients. PMID- 15894899 TI - Acute pressure-natriuresis function shows early impairment in Lyon hypertensive rats. AB - OBJECTIVE: This study aimed to determine whether the alteration of the pressure natriuresis seen in Lyon genetically hypertensive (LH) rats occurs early, and the possible involvement in this alteration of the most important extra-renal factors that influence natriuresis. METHODS: In LH rats and their normotensive (LL) controls, acute pressure natriuresis was studied in denervated kidneys with or without controlling extra-renal influence; that is, adrenalectomy and an intravenous infusion of vasopressin, norepinephrine, hydrocortisone and aldosterone. RESULTS: With controlling the cited extra-renal influence, LH rats already exhibited, at 5 weeks of age, a slightly higher blood pressure (+9%) and a markedly reduced renal blood flow (-33%) compared with LL rats; their pressure diuresis and pressure-natriuresis curves were significantly blunted. Between 16 and 50 weeks of age, although BP levels did not change, renal blood flow and glomerular filtration rate declined in LH rats while their pressure-diuresis and pressure-natriuresis curves continued to shift to higher pressures. When studied without controlling extra-renal influence, the values of pressure diuresis and natriuresis were significantly higher than in controlled conditions both in LH and LL rats. However, in 16-week-old rats, the LH/LL ratios for sodium and water excretion remained close under the two experimental conditions. CONCLUSIONS: The pressure-natriuresis function in LH rat shows early impairment and aggravates with age. This alteration is observed with, as well as without, controlling the influence of the main extra-renal factors that affect natriuresis. PMID- 15894900 TI - Long-term exercise training and angiotensin-converting enzyme inhibition differentially enhance myocardial capillarization in the spontaneously hypertensive rat. AB - OBJECTIVES: To investigate whether combined treatment with lisinopril, an angiotensin-converting enzyme (ACE) inhibitor and exercise training would have an additive effect in enhancing the capillary supply of the left ventricular (LV) myocardium in spontaneously hypertensive rats (SHR). DESIGN: Twelve-week-old male SHR were divided into four groups (10-12 each): sedentary, sedentary treated with lisinopril (15-20 mg/kg per day by gavage), exercise trained, and exercise trained while treated with lisinopril. Exercise training consisted of 1 h a day/5 days a week of running on a treadmill. METHODS: After 10 weeks of experimental protocols, capillary surface density and length density were sterologically determined in 1 mum thick LV tissue samples from perfuse-fixed hearts. RESULTS: Lisinopril significantly reduced systolic blood pressure (SBP) and LV mass in the sedentary with lisinopril and exercise trained with lisinopril groups but did not affect the heart rate (HR). Exercise training did not reduce SBP or LV mass, but significantly reduced HR in the exercise trained and exercise trained with lisinopril groups. Lisinopril treatment (sedentary with lisinopril), exercise training (exercise) and their combination (exercise trained with lisinopril) significantly increased myocardial capillary surface area density by 26, 38 and 65% and length density by 38, 48 and 67%, respectively. CONCLUSION: Lisinopril administration and exercise training independently enhanced myocardial capillarization through a reduction of myocardial mass and stimulation of angiogenesis, respectively. A combination of the two treatments enhanced myocardial capillarization more than either intervention alone. This may aid in the restoration of the normal nutritional status of cardiac myocytes compromised by the hypertrophic state of hypertension. PMID- 15894901 TI - Effects of a lifestyle programme on ambulatory blood pressure and drug dosage in treated hypertensive patients: a randomized controlled trial. AB - OBJECTIVE: To assess effects of multifactorial lifestyle modification on antihypertensive drug needs in treated hypertensive individuals. DESIGN: Randomized controlled trial. SETTING: Research studies unit. PARTICIPANTS: Overweight hypertensive patients, receiving one or two antihypertensive drugs, were recruited by advertising, and allocated randomly to a usual care group (controls; n = 118) or a lifestyle modification group (programme group; n = 123). INTERVENTION: A 4-month programme of weight loss, a low-sodium 'Dietary Approaches to Stop Hypertension'-type diet with added fish, physical activity and moderation of alcohol intake. After 4 months, if mean 24-h ambulatory blood pressure (ABP) was less than 135/85 mmHg, antihypertensive drugs were withdrawn over 4 weeks and long-term home blood pressure monitoring was begun. MAIN OUTCOME MEASURES: Antihypertensive drug requirements, ABP, weight, waist girth at 4 months and 1-year follow-up. RESULTS: Ninety control group and 102 programme group participants completed the study. Mean 24-h ABP changed after 4 months by 1.0/-0.3 +/- 0.5/0.4 mmHg in controls and -4.1/-2.1 +/- 0.7/0.5 mmHg with the lifestyle programme (P < 0.01). At follow-up, changes in the two groups were not significantly different (4.1/1.3 +/- 1.1/1.0 mmHg in controls; 2.5/-0.1 +/- 1.1/0.8 mmHg in the programme group; P = 0.73). At 4 months, drug withdrawal differed significantly between the groups (P = 0.038) in men (control 44%; programme 66%) but not in women (65 and 64%, respectively; P = 0.964). At follow up, sex-related differences were not significant, and 41% in the control group and 43% in the programme group maintained drug-withdrawal status. With the programme, net weight loss was 3.3 kg (P < 0.001) at 4 months and 3.0 kg (P < 0.001) at follow-up; respective net decreases in waist girth were 3.3 cm (P < 0.001) and 3.5 cm (P < 0.001). CONCLUSIONS: A 4-month multifactorial lifestyle modification in patients with treated hypertension reduced blood pressure in the short-term. Decreased central obesity persisted 1 year later and could reduce overall cardiovascular risk. PMID- 15894902 TI - Effect of the renin-angiotensin system or calcium channel blockade on the circadian variation of heart rate variability, blood pressure and circulating catecholamines in hypertensive patients. AB - OBJECTIVE: To determine the effects of 8 weeks of therapy with amlodipine, ramipril or telmisartan on the autonomic system over 24 h in hypertensives. METHODS: After a placebo run-in, 57 patients were included in a prospective randomized open-label design protocol for therapy with amlodipine (5 mg for 4 weeks followed by 10 mg for 4 weeks, n = 22), or ramipril (2.5 mg for 1 week, 5.0 mg for 3 weeks and 10 mg for 4 weeks, n = 17) or telmisartan (80 mg for 8 weeks, n = 18). Autonomic functions were assessed by norepinephrine (NE) and epinephrine (E), as well as by the spectral analysis of heart rate variability (HRV). RESULTS: The 24-h ambulatory blood pressure, plasma NE and HRV demonstrated the characteristic day-night circadian rhythm in hypertensives. Higher values for SBP and DBP and for NE levels, as well as for spectral analysis components - low frequency band (LF) and low frequency/high frequency (LF/HF) ratio - were found during the day, whereas the HF was higher during the night. In patients treated with amlodipine, the HF decreased significantly during the night, while the LF and the LF/HF ratio increased during the day in association with the rise in NE. The therapy with telmisartan did increase the HF during the night and the day, while ramipril did not influence all HRV components during the night but significantly increased the HF, and decreased the LF/HF ratio during the day. No changes were observed in plasma NE with telmisartan or ramipril, but a 50% increase in NE levels throughout the 24-h period was found in amlodipine-treated patients. CONCLUSION: These data suggest a sympathetic activation during the day and a decrease in parasympathetic activity during the night after therapy with amlodipine, correlated with increases in plasma NE. In contrast, the therapy with telmisartan significantly increased parasympathetic activity without changes in NE during the night and day. The therapy with ramipril increased the parasympathetic activity only during the day. PMID- 15894903 TI - Is interview a reliable method to verify the compliance with antihypertensive therapy? An international central-European study. AB - BACKGROUND: Non-compliance with prescribed antihypertensive medication is an important contributor to the failure of antihypertensive therapy. OBJECTIVE: To assess the validity of a short questionnaire in the identification of non compliant patients. METHODS: In three central-European countries, work-site screening for hypertension was conducted. Blood pressure was measured using an automatic electronic blood pressure measuring device (BpTRU). Respondents were interviewed by trained personnel and a short questionnaire focused on blood pressure awareness and treatment compliance was completed. RESULTS: A total of 2812 persons were screened: 841(29.9%) respondents were hypertensive, and out of these the total number of treated hypertensive subjects was 359 (42.6%). Mean systolic blood pressure and diastolic blood pressure were significantly lower in the compliant group than the non-compliant group (systolic blood pressure, 139.4 and 146.2 mmHg, respectively, P = 0.002; and diastolic blood pressure, 89.2 and 92.3 mmHg, respectively, P < 0.01). The non-compliant group was younger than the compliant group (mean age, 46.7 versus 48.9 years, respectively, P = 0.01). Females, patients on combined therapy and non-smokers were more compliant than males, those on mono-therapy and smokers (P = 0.01, P = 0.004 and P = 0.005, respectively). CONCLUSION: Patients reporting strict compliance with prescribed drug therapy have significantly lower systolic blood pressure and diastolic blood pressure than those who admit even an occasional lapse in taking medication. A properly formulated questionnaire can identify non-compliant patients. PMID- 15894904 TI - Alberto C. Taquini and the 'links' that led to the discovery of angiotensin: on the 100th anniversary of his birth. PMID- 15894905 TI - The relationship between baseline blood pressure and magnitude of postexercise hypotension. PMID- 15894907 TI - Bartter's and Gitelman's syndromes: a confirm in humans of the utility of Rho kinase inhibition for cardiovascular protection. PMID- 15894908 TI - The prevalence of hypertension in Persian Gulf countries and its correlation with demographic and socio-economic factors. PMID- 15894912 TI - Image metrics for predicting subjective image quality. AB - PURPOSE: Despite the proliferation of wavefront sensors to characterize the optical quality of individual eyes, there is not yet an accurate way to determine from a wave aberration how severely it will impact the patient's vision. Some of the most commonly used metrics, such as RMS wavefront error and the Strehl ratio, predict subjective image quality poorly. Our goal is to establish a better metric to predict subjective image quality from the wave aberration. METHODS: We describe three kinds of experiments designed to compare the effectiveness of different metrics in determining the subjective impact of the wave aberration. Subjects viewed a visual stimulus through a deformable mirror in an adaptive optics system that compensated for the subject's wave aberration. In the first experiment, we show that some Zernike modes such as spherical aberration and defocus interact strongly in determining subjective image quality. In the second experiment, the subject's wave aberration was replaced by the wave aberration corresponding to an individual Zernike mode. The subject then adjusted the coefficient of the Zernike mode to match the blur of a standard stimulus. In the third experiment, the subject viewed the same stimulus through the wave aberration of one of 59 different postoperative patients who had undergone LASIK and matched the blur by adjusting defocus. We then determined which among many image quality metrics best predicted these matching data. RESULTS: RMS wavefront error was a poor predictor of the data, as was the Strehl ratio. CONCLUSIONS: The neural sharpness metric best described the subjective sharpness of images viewed through the wave aberrations of real eyes. This metric can provide a single number that describes the subjective impact of each patient's wave aberration and will also increase the accuracy of refraction estimates from wavefront-based autorefractors and phoropters. PMID- 15894913 TI - Short-term effects of artificial tears on visual performance in normal subjects. AB - PURPOSE: Temporal changes in tear film structure can distort the optical wavefront as it passes through the tear layer and reduce contrast sensitivity. Theoretically, any substance applied to the tear layer that alters its structure could affect contrast sensitivity. The purpose of this study is to investigate how different formulations of carboxymethylcellulose sodium (CMC) applied to the tear layer affect contrast sensitivity over time. Additionally, the visual effect of these drops applied over soft and rigid, gas-permeable contact lenses was also investigated. METHODS: Twenty normal subjects took part in this project. Refresh Celluvisc (Allergan, Irvine, CA, 1.0% high-viscosity CMC) was compared with Refresh Liquigel (Allergan, 1.0% total CMC made by blending 0.35% high-viscosity with 0.65% medium viscosity CMC). Ten of the subjects were habitual soft contact lens wearers and 10 were habitual gas-permeable lens wearers. The stimulus, viewed monocularly, was a stationary, vertically oriented, sine wave grating (14 CPD). A temporal, two-alternative, forced-choice paradigm combined with a self paced method of limits was used to monitor threshold over time. After baseline data collection, a drop of the artificial tear was applied to the tear layer and the procedure continued for 30 min. This allowed continual tracking of the threshold. Data were collected while viewing the stimulus with the subject's contact lens or with their spectacle prescription. RESULTS: One drop of Liquigel or Celluvisc decreased contrast sensitivity for a 14 CPD sine wave grating (all p values < 0.005). This decrease in contrast sensitivity was observed during spectacle, soft contact lens, and gas-permeable contact lens wear. Soft contact lens wear resulted in a greater decrease in contrast sensitivity than spectacles when Liquigel was applied to the tear layer. None of the other conditions were different between contact lens and spectacle wear. The return to baseline contrast sensitivity was not significantly different between soft or gas permeable contact lens wear and spectacles for either Liquigel or Celluvisc. In general, Celluvisc had a greater effect on visual performance than Liquigel. CONCLUSIONS: These results suggest that Liquigel and Celluvisc alter the tear layer and affect contrast sensitivity. The results agree with patient observations that Celluvisc causes a moderate amount of blur that gradually subsides. In such patients, the shorter duration of blur with Liquigel (about half that of Celluvisc) may be more acceptable. The technique of blending various viscosity CMC materials while maintaining the total CMC concentration of 1.0% may be beneficial in dry eye therapy without causing excessive blur to patients. PMID- 15894914 TI - Spectral absorbance and spatial distribution of macular pigment using heterochromatic flicker photometry. AB - PURPOSE: The validity of the heterochromatic flicker photometric (HFP) technique for measuring macular pigment (MP) was assessed by evaluating the spatial density and spectral absorption curves of normal subjects. METHODS: Standard Maxwellian view optics were used to measure MP spatial density and spectral absorption curves. Four subjects with normal color vision and one protanope were tested. All of the subjects were experienced psychophysical observers. RESULTS: Spatial density profiles for all subjects were nearly symmetric. The width of the spatial distribution, at half maximal density, averaged 0.70 degrees (standard deviation = 0.15). A first-order exponential decay with eccentricity described the distribution profile well (average r = 0.95). The shape of the spectral absorption curves matched an ex vivo template closely at wavelengths greater than approximately 430 nm. Stimulus size did not influence the derived spectral curves. CONCLUSION: Based on data from this study and others, the HFP method appears to be a valid method for measuring MP density in subjects without retinal disease. PMID- 15894915 TI - Assessment of the validity of in vivo methods of measuring human macular pigment optical density. AB - PURPOSE: As a result of the putative protective role of lutein and zeaxanthin at the site of the retina (macular pigment [MP]), interest in the noninvasive measurement of these retinal carotenoids is increasing. In principle, any method used to measure MP in vivo should provide spectral absorption curves that match extinction spectra of MP measured ex vivo. METHODS: In this article, we assess published spectral curves with respect to this criterion. RESULTS AND DISCUSSION: Variation in derived MP measures resulting from methodological differences between techniques is noted and discussed. PMID- 15894916 TI - Repeatability of corneal topography measurement in keratoconus with the TMS-1. AB - PURPOSE: The purpose of this study was to report the test-retest variability of simulated indices derived from the TMS-1 topography instrument (Tomey Technology, Waltham, MA) in keratoconus subjects enrolled in the Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study. METHODS: Four images were taken at an initial visit and at a repeat visit several weeks later. From these images, 17 indices were simulated from published formulas. Mixed-model analysis was used on test-retest data from the TMS-1 videokeratography instrument during the baseline year. This analysis yields estimates of within- and between-visit variability. RESULTS: Repeatability analysis revealed that within-visit standard errors were 1.0 to 5.9 times greater in keratoconus eyes than in normal controls when two images were analyzed from each visit. These values changed only slightly when more images were used. The ratio of between-visit standard errors of the indices were nearly equally greater than normal controls for (0.9-4.6 and 0.9-4.3) two images per eye and all images per eye, respectively. CONCLUSIONS: These results suggest that the repeatability of simulated indices derived from TMS-1 topography in keratoconus subjects is poorer than in normal controls. PMID- 15894917 TI - Comparison of human central cornea and limbus in vivo using optical coherence tomography. AB - PURPOSE: The purpose of this study was to compare central corneal and limbal total and epithelial thickness using a commercially available optical coherence tomographer. METHODS: A Humphrey-Zeiss Optical Coherence Tomographer (OCT [Carl Zeiss, Meditec, Dublin, CA]) was used to obtain corneal images from 10 subjects. Central corneal and limbal total and epithelial thickness of both eyes were measured using the OCT. Each OCT image comprised 100 measurements, 10 nasal, 10 central, and 10 temporal measurements from each image were analysed. RESULTS: The central corneal and epithelial thickness of the right and the left eyes were 507.9 +/- 35.8 microm, 58.4 +/- 2.5 microm, 506.9 +/- 37.4 microm, and 58.5 +/- 2.5 microm, respectively. There were no differences between eyes (p > 0.05). The nasal and temporal limbal total and epithelial thickness of the right and left eyes were 703.8 +/- 32.1 microm, 704.9 +/- 31.0 microm, 76.8 +/- 3.5 microm, 77.9 +/- 2.9 microm, 704.4 +/- 31.8 microm, 706.3 +/- 32.5 microm, 77.5 +/- 2.8 microm, and 77.8 +/- 2.5 microm, respectively. There were no differences between the nasal and temporal total and epithelial thickness of both eyes (p > 0.05). However, there was a statistical difference between the central corneal and limbal total and epithelial thickness (both p < 0.05). CONCLUSIONS: Central cornea and limbus are measurably different using OCT. Central cornea is thinner than limbus for both total thickness and epithelial thickness. There is no difference between eyes of central corneal and limbal total and epithelial thickness. Optical Coherence Tomography is a useful instrument for in vivo human limbal morphometry. PMID- 15894918 TI - A comparative study of the performance of different corneal topographers on children with respect to orthokeratology practice. AB - PURPOSE: The purpose of this study was to evaluate and compare the performance of three topographers, Medmont E300, Keratron Scout, and Humphrey Atlas 991, for the measurement of corneal parameters of interest in orthokeratology practice on young children. METHODS: Apical radius (R0), flattest corneal curvature (flat k), eccentricity (e) value, and elevation at 9-mm chord were measured twice on 22 healthy subjects of mean +/- standard deviation (SD) age 11.2 +/- 2.2 years using the topographers under investigation. RESULTS: The repeatability of the Medmont and Humphrey were good for the measurements of R0 and flat k. The repeatability of the Humphrey was also good for the measurement of e-value. However, for both topographers, the measurement of elevation was not precise enough for orthokeratology empirical lens design. The Keratron showed poor repeatability for all the parameters evaluated. The agreement of the parameters of interest between these instruments was poor. CONCLUSIONS: The Medmont and the Humphrey showed good repeatability on children, but the precision of elevation measurements was not good enough to be used for designing empiric orthokeratology lenses on children. The Keratron is not recommended to be used on children in orthokeratology fitting and management. Interinstrumental agreements were significantly different and therefore measurements from different topographers should not be used interchangeably. PMID- 15894919 TI - High interocular corneal symmetry in average simulated keratometry, central corneal thickness, and posterior elevation. AB - PURPOSE: The purpose of this study was to assess interocular corneal symmetry in average simulated keratometry, corneal thickness, and posterior corneal elevation. METHODS: This retrospective analysis included data from scanning slit topography (Orbscan II; Bausch and Lomb, Rochester, NY) on 242 eyes from 121 consecutive patients undergoing standard evaluation for consideration of elective laser vision correction. The symmetry between the right and left eye in average simulated keratometry, minimum central corneal thickness, and posterior corneal elevation was assessed by comparative data analysis. RESULTS: Simulated keratometry ranged from 39.9 to 48.6 D. The interocular difference in average simulated keratometry was 0.47 D (standard deviation [SD] 0.43). The interocular Pearson correlation coefficient for average simulated keratometry was 0.90 (p < 0.001). The range of minimum corneal thickness was 432 to 628 microm. The interocular Pearson correlation coefficient for minimum central corneal thickness was 0.95 (p < 0.001). Right and left eye minimum corneal thickness differed by an average 8 microm (SD 7). The range of posterior elevation was -4 to 54 microm. The average difference in posterior corneal elevation between the right and left eye was 6 microm (SD 5). The interocular Pearson correlation coefficient for posterior corneal elevation was 0.72 (p < 0.001). The average posterior elevation was 19 microm (SD 11). CONCLUSIONS: Although a wide range of values exists in simulated keratometry, minimum corneal thickness, and posterior corneal elevation, interocular symmetry in all these parameters was very high in this group of consecutive patients. Asymmetry of these interocular parameters may warrant repeat clinical testing for accuracy and may predict corneal abnormalities. Normative data on posterior cornea elevation is presented. This study points out potentially clinically important high interocular corneal symmetry data in simulated keratometry, corneal thickness, and posterior corneal elevation. PMID- 15894920 TI - Sensitivity of screening tests for detecting vision in preschoolers-targeted vision disorders when specificity is 94%. AB - PURPOSE: To compare the sensitivity of 11 preschool vision screening tests administered by licensed eye care professionals for the detection of the 4 Vision in Preschoolers (VIP)-targeted vision disorders when specificity is 94%. METHODS: This study consisted of a sample (n = 2588) of 3- to 5-year-old children enrolled in Head Start programs, 57% of whom had failed an initial Head Start vision screening. Screening results from 11 tests were compared with results from a standardized comprehensive eye examination that was used to classify children with respect to the four VIP-targeted vision disorders: amblyopia, strabismus, significant refractive error, and unexplained reduced visual acuity (VA). With overall specificity set to 94%, we calculated the sensitivity for the detection of each targeted vision disorder. RESULTS: With the overall specificity set to 94%, the most accurate tests for detection of amblyopia were noncycloplegic retinoscopy (NCR) (88% sensitivity), the SureSight Vision Screener (80%), and the Retinomax Autorefractor (78%). For detection of strabismus, the most accurate tests were the MTI Photoscreener (65%), the cover-uncover test (60%), the Stereo Smile II stereoacuity test (58%), the SureSight Vision Screener (54%), and the Retinomax Autorefractor (54% in year 1, 53% in year 2). The most accurate tests for detection of significant refractive error were NCR (74%), the Retinomax Autorefractor (66%), the SureSight Vision Screener (63%), and the Lea Symbols VA test (58%). For detection of reduced VA, the most accurate tests were the Lea Symbols Distance VA test (48%), the Retinomax Autorefractor (39%), and NCR (38%). CONCLUSIONS: Similar to the previously reported results at 90% specificity, the screening tests vary widely in sensitivity with specificity set at 94%. The rankings of the sensitivities for detection of the 4 VIP-targeted vision disorders are similar to those with specificity set to 90%. PMID- 15894921 TI - T-cell/histiocyte-rich large B-cell lymphoma displays a heterogeneity similar to diffuse large B-cell lymphoma: a clinicopathologic, immunohistochemical, and molecular study of 30 cases. AB - T-cell/histiocyte-rich large B-cell lymphoma (THRLBCL), a proliferating peripheral B-cell neoplasm, is a morphologic variant of diffuse large B-cell lymphoma (DLBCL), which may be confused with Hodgkin's lymphoma, non-Hodgkin's lymphoma, and reactive lymphadenopathies. Though more recent studies suggested that it might be a distinct clinicopathologic entity and/or a heterogeneous entity with derivation from germinal center B cells, its histogenetic derivation remains controversial. The authors analyzed 30 cases of THRLBCL to further characterize the origin of the neoplastic cells using immunohistochemical and molecular studies for expression of Bcl-6, CD10, and CD138, as well as rearrangements of IgH/bcl-2 genes on paraffin-embedded tissue. Half of the cases (15/30) showed Bcl-6 expression and five cases (19%) showed CD10 expression, but none had CD138 expression (0/20). Only three cases showed coexpression of both Bcl-6 and CD10. Molecular studies performed in 21 cases detected rearrangement of immunoglobulin heavy gene in 18 cases, with none having detectable Bcl-2 gene rearrangement. These data indicate that similar to DLBCL, the cell origin of neoplastic cells in THRLBCL is composed of a heterogeneous group of proliferating peripheral B cells, with only some cases originating from germinal center B cells and others derived from heterogeneous origins. Lack of Bcl-2 gene rearrangements seems to argue against a possible progression from preexisting follicular lymphoma. Thus, the normal counterpart of the neoplastic cells cannot at this time be the sole basis for the subclassification of THRLBCL. PMID- 15894922 TI - The t(14;18) in diffuse large B-cell lymphoma: correlation with germinal center associated markers and clinical features. AB - The clinical and biologic relevance of the t(14;18) and features of germinal center (GC) differentiation in diffuse large B-cell lymphoma (DLBCL) remain controversial. The authors examined the association of t(14;18) with GC associated markers and clinical features in 44 de novo DLBCLs (22 nodal and 22 primary extranodal). CD10, bcl-2, and bcl-6 were expressed in 50%, 62%, and 54% of cases respectively. There were no significant differences in expression of these markers between nodal and extranodal cases. Coexpression of CD10 and bcl-6 was seen in 12 of 41 cases, and was more frequent in nodal than extranodal DLBCL (9 of 21 vs. 3 of 20; P = 0.05). A CD10+/bcl-6+ phenotype was not significantly associated with bcl-2 expression, stage, complete remission rate, or survival. The t(14;18) was found in 7 of 44 (16%) cases (6 nodal, 1 extranodal; P = 0.09). It was associated with a CD10+/bcl-6+ phenotype (5 of 7 vs. 7 of 27; P = 0.015) and a trend toward more frequent bcl-6 expression (6 of 7 vs. 15 of 34; P = 0.09), but no association with bcl-2 expression, CD10, clinical stage, complete remission, or survival. Among nodal or high-stage (III-IV) DLBCL, cases with the t(14;18) showed a trend toward decreased survival (P = 0.12). PMID- 15894923 TI - Centrocytoid plasma cells of the germinal center. AB - Germinal centers within the lymph node follicles are T-cell-dependent, antigen driven B-cell proliferations that develop from the rapid clonal expansion of a few founder cells. The end results of this B-cell expansion are memory B cells or plasma cells. Two morphologic forms of plasma cell can be recognized in the germinal center: classic plasma cells, characterized morphologically by peripherally clumped arrangement of nuclear chromatin, and cells with a nuclear morphology more resembling that of the centrocytes, which the authors have termed "centrocytoid plasma cells." In this study the authors examined the distribution and immunohistochemical characteristics of these two populations of germinal center plasma cells. The centrocytoid plasma cells were arranged in a band stretching from the junction of the dark and light zone to the periphery of the germinal centers, while the classic plasma cells were mainly present at the germinal center periphery. Both marked with CD38, CD138, and VS38c, recognized markers for plasma cells; however, EMA and CD31 were present only in the classic form of plasma cell. The proliferation marker Ki67 was present in less than 1% of the cells labeling with CD138. Others have demonstrated Ki67 in 50% of the cells labeled with Blimp-1, which is consistent with Blimp-1 appearing earlier than CD138 in ontogeny. CD10 is co-expressed with CD138 in about 10% of cells and CD45 with CD138 in about 5% of cells. Their topographic features, together with the progressive acquisition of plasma cell markers, suggest that the centrocytoid plasma cells may be the precursors of the classic plasma cells. Of note, both the forms of plasma cell were absent in follicles of follicular lymphoma, which supports the concept that in this disease, lymphocytes fail to differentiate and mature beyond the centrocyte stage. PMID- 15894924 TI - Differential expression of WT1 gene product in non-Hodgkin lymphomas. AB - The tumor suppressor gene wt1 (Wilms tumor 1) encodes a zinc finger transcription factor reported to be expressed in many tumors, including mesotheliomas, carcinomas, and acute leukemias. However, WT1 expression in non-Hodgkin lymphomas (NHLs) has not been studied. The authors assessed for WT1 expression in six lymphoma/leukemia cell lines using Western blot methods after subcellular fractionation. We also assessed for WT1 expression in 167 NHLs using immunohistochemical methods. The B-cell NHLs analyzed were 18 diffuse large B cell lymphomas, 13 marginal zone B-cell lymphomas, 9 small lymphocytic lymphomas, (DLBCLs), 8 follicular lymphomas, 6 mantle cell lymphomas, 5 Burkitt lymphomas, 3 lymphoplasmacytic lymphomas, and 2 B-cell lymphoblastic lymphomas. The T-cell NHLs analyzed were 43 anaplastic large cell lymphomas (ALCLs), 26 peripheral T cell lymphomas unspecified, 13 angioimmunoblastic T-cell lymphomas, 6 cutaneous ALCLs, 6 cases of mycosis fungoides, 5 extranodal NK/T-cell lymphomas of nasal type, and 4 T-cell lymphoblastic lymphomas. WT1 levels were higher in cytoplasmic extracts than in nuclear extracts of the Karpas 299 and SU-DHL-1 lymphoma cell lines but were higher in nuclear extracts than in the cytoplasmic extracts of the Jurkat, HH, U-937, and K562 leukemia cell lines. In NHLs, WT1 was positive in 4 of 5 (80%) Burkitt lymphomas, 9 of 12 (75%) ALK-positive ALCLs, 3 of 6 (50%) lymphoblastic lymphomas (2 of 4 T-cell, 1 of 2 B-cell), 14 of 31 (45%) ALK negative ALCLs, 6 of 18 (33%) DLBCLs, and 1 of 6 (17%) cutaneous ALCLs. WT1 was negative in all other NHLs tested. WT1 immunoreactivity was primarily cytoplasmic in all positive NHLs except T-cell lymphoblastic lymphoma. In conclusion, WT1 protein is frequently detected in the cytoplasm of a subset of high-grade NHLs. PMID- 15894925 TI - Coexpression of CD43 by benign B cells in the terminal ileum. AB - Expression of CD43 by B cells is often used as a diagnostic criterion in favor of a B-cell lymphoproliferative disorder, including small lymphocytic lymphoma/chronic lymphocytic leukemia, mantle cell lymphoma, Burkitt lymphoma, precursor B-lymphoblastic lymphoma, and a subset of marginal zone B-cell lymphomas. Benign B cells generally do not coexpress CD43. The authors analyzed 20 biopsies of the terminal ileum for nonneoplastic disease for expression of CD43 and compared them with other sites and with CD20, CD138, and CD3 reactivity. The majority of cases (85%) showed strong coexpression of CD43 by benign perifollicular B cells. The presence of CD43 coexpression in B-cell populations of the terminal ileum, including those of Peyer's patches, should not be used as a diagnostic parameter to differentiate extranodal marginal zone B-cell lymphoma of MALT type from reactive processes. PMID- 15894926 TI - Expression of CDX2, cytokeratins 7 and 20 in sinonasal intestinal-type adenocarcinoma. AB - CDX2 is a transcription factor expressed by intestinal epithelium. It is considered as a sensitive marker for a colorectal or-less frequently-gastric origin of adenocarcinomas. The pattern of coordinated expression of cytokeratin (CK) 7 and CK20 is also useful for the diagnosis of the origin of metastatic adenocarcinomas. Expression of CDX2, CK7 and CK20 was investigated in 14 cases of sinonasal intestinal-type adenocarcinoma (SIA), a particular tumor with an enteric-cell-type appearance. Formalin-fixed, paraffin embedded tissue sections were immunostained with monoclonal antibodies using the biotin-labeled streptavidin technique. All of the cases expressed CDX2, being stained 50 to 100% of the tumor cells (mean: 87.2%). CK7 positivity was detected in 8 cases (57.1%), with the staining of 10 to 100% of cells (mean: 65.6%), and CK20 was found in all the tumors (10 to 100% of cells; mean: 78.8%). The histologic resemblance between SIA and colorectal adenocarcinoma is reinforced by the expression of CDX2 and CK20, which are virtually constant in both neoplasms. CDX2 seems to be a marker for cellular phenotype better than an indicator of the origin of adenocarcinomas. CK7 is expressed in SIA less frequently than in colonic adenocarcinoma, but with a rate similar to the subset of rectal tumors, making the differential diagnosis between the two neoplasms difficult. PMID- 15894927 TI - Expression of vascular endothelial growth factor and proliferation marker MIB1 are influenced by neoadjuvant chemotherapy in locally advanced breast cancer. AB - Neoadjuvant chemotherapy (NACT) has become the standard of care for patients with locally advanced breast cancer (LABC). This was a retrospective review of 21 consecutive women who received NACT as initial treatment of LABC, followed by surgical excision. The pre- and post-treatment breast specimens and post treatment axillary lymph nodes with metastases were immunostained to evaluate for proliferative index (PI) (MIB-1 Immunotech) and vascular endothelial growth factor (VEGF) expression (Santa Cruz, CA, clone A-20). Thirteen of the 21 patients (62%) had more than 50% tumor shrinkage following NACT. The breast's mean PI decreased from 47.86% to 23.95% after treatment (P = 0.005). The mean PI in the post-treatment lymph nodes was 24.47%. A nodal post-NACT PI of less than 10% and progesterone receptor-positive tumor status were associated with better survival, as all such patients are alive. A high PI after NACT was associated with recurrence or death. All of the patients who showed an excellent clinical response had either a decrease in the PI or an absence of a high level of VEGF after NACT. Most patients exhibited persistent expression of VEGF after NACT. Pathologic response in the primary tumor did not correlate with the response in the lymph nodes or with overall survival. NACT reduces the size and PI of the primary breast tumor independent of the patient's node status. The PI may be an early means by which to identify tumors most likely to reduce in size with chemotherapy. A low PI after NACT is associated with better survival. There is persistent expression of VEGF in post-NACT residual breast carcinoma. Thus, anti VEGF drugs after conventional chemotherapy may benefit patients with residual carcinoma. PMID- 15894928 TI - Epithelioid gastrointestinal stromal tumor with PDGFRA activating mutation and immunoreactivity. AB - The authors report a unique case of an intra-abdominal, epithelioid mesenchymal tumor that had an activating mutation of PDGFRA and a strong PDGFRA immunoreactivity but lacked both c-kit mutation and c-kit protein (CD117) expression. IHC study showed that the tumor cells were diffusely and strongly positive for PDGFRA, vimentin, CD34, and Bcl-2 but completely negative for CD117 as well as for muscle, epithelial, endothelial, endocrine, mesothelial, neural, and melanocytic cell markers. Molecular study revealed a mutation at the juxtamembrane domain of exon 12 in PDGFRA gene with GTC to GAC transition at codon 561 (V561D), as shown in the previous mutational studies on gastrointestinal stromal tumor (GIST). This case likely represents an example of GIST with PDGFRA activating mutation and PDGFRA immunoreactivity without CD117 positivity, which has not been documented in the literature. STI 571 (imatinib mesylate [Gleevec]) might be an effective therapy in this case, since Gleevec targets both PDGFRA and c-kit oncoproteins. PMID- 15894929 TI - C-Kit (CD117) expression in AIDS-related, classic, and African endemic Kaposi sarcoma. AB - Kaposi sarcoma (KS) is a multicentric vascular neoplasm characterized histologically by the progressive proliferation of spindle-shaped tumor cells in all epidemiologic (AIDS-related, classic, endemic, and iatrogenic) forms. Human herpesvirus 8 (HHV8) is associated with all epidemiologic forms of KS and has been shown in vitro to induce the tyrosine receptor kinase c-kit in HHV8-infected cells. To date, c-kit immunoreactivity has not been systematically studied in KS lesions. Therefore, the aim of this study was to evaluate c-kit expression by immunohistochemistry in different proliferative stages and epidemiologic forms of KS. Archival cases of formalin-fixed, paraffin-embedded KS lesions, including 9 classic, 11 AIDS-related, and 15 African (endemic) forms at various histologic stages (5 patch, 8 plaque, 22 nodular), biopsied from different sites, were stained using immunohistochemistry with antibodies to HHV8 (LNA-1) and c-kit (CD117). C-kit immunoreactivity of lesional cells was demonstrated in 15 (43%) cases overall. A total of five (56%) classic, five (45%) AIDS-related, and five (33%) endemic KS cases were positive for c-kit. There was no difference in the intensity of c-kit staining between the different epidemiologic groups and histologic stages of KS. HHV8 (LNA-1) immunoreactivity was present in all (100%) classic, 10 (91%) AIDS-related, and 9 (60%) endemic cases. LNA-1 staining was demonstrated in 13 (93%) of the c-kit-positive and 15 (75%) of the c-kit-negative KS lesions. These findings indicate that c-kit expression in lesional cells can be detected by immunohistochemistry in different epidemiologic forms and histologic stages of KS. Furthermore, the expression of c-kit does not correspond with the presence of HHV8 (LNA-1) immunoreactivity in KS lesions. PMID- 15894930 TI - Imatinib mesylate (gleevec)--targeted kinases are expressed in uterine sarcomas. AB - The purpose of this study was to determine whether 3 tyrosine kinases known to be inhibited by imatinib mesylate are expressed in a variety of uterine sarcomas. The authors assessed c-kit, abl, and platelet-derived growth factor receptor-beta (PDGFR-beta) expression in 8 endometrial stromal sarcomas (ESSs), 5 leiomyosarcomas (LMSs), 4 high-grade endometrial sarcomas (HGESs), and 21 malignant mixed mullerian tumors (MMMTs). Tissue sections were stained with commercially available antibodies for c-kit, abl, and PDGFR-beta. Staining intensity was described as 0 (no staining), +1 (weak), +2 (moderate), and +3 (strong). Positive staining was defined as moderate to strong if found in more than 10% of tumor cells. Expression of c-kit ranged from 0% in LMSs to 25% in HGESs. Protein expression of abl was more significant, ranging from 25% in LMSs and ESSs to 43% in MMMTs. Only 1 LMS sample stained focally for abl (+1). Abl expression was observed in only the carcinomatous elements of the MMMTs, with diffuse staining in the cytoplasm and nucleus. In most, the staining intensity was +2. All tumors stained positive for PDGFR-beta. MMMT samples showed PDGFR beta expression in both the carcinomatous and sarcomatous portions. In all samples, staining for PDGFR-beta was concentrated at the cell membrane and diffusely in the cytoplasm. These results indicate that many uterine sarcomas express 1 or more of the kinases targeted by imatinib mesylate and that further investigation of imatinib as a therapy for uterine sarcomas is warranted. PMID- 15894931 TI - Diagnostic utility of cell cycle and apoptosis regulatory proteins in verrucous squamous carcinoma. AB - A major problem in the diagnosis of verrucous squamous cell carcinoma is the lack of readily reproducible objective criteria for distinguishing this malignant lesion from reactive epithelial hyperplasia. Both lesions are characterized by thickened (well-differentiated) squamous epithelium without cellular atypia and subjacent stroma densely infiltrated by lymphocytes and plasma cells. This study was carried out to evaluate the use of cell cycle and apoptosis-related regulatory proteins in the diagnosis of verrucous carcinoma. The study materials consisted of representative formalin-fixed and paraffin-embedded tissue blocks from 19 cases of verrucous carcinoma, 18 classic squamous cell carcinoma, and 14 squamous epithelial hyperplasia (acanthosis). The immunohistochemical expression of the following of cell cycle and apoptosis-related regulatory proteins was evaluated using avidin-biotin complex detection technique: p16, p21, p53, Ki67, and retinoblastoma gene product (RBGP) (also known as retinoblastoma protein [pRb]). Expression of Ki67 was detected only in the single basal layer of the epithelium in all 14 cases of acanthosis. In verrucous carcinoma, Ki67 was detected in basal and suprabasal cells in the lower third of the neoplastic epithelium in 19 of 19 cases (100%). In neoplastic squamous epithelium with frankly invasive squamous cell carcinoma, Ki67 was diffusely expressed throughout the entire thickness of the epithelium as well as in the underlying invasive tumor nests. The pattern of p53 expression was similar to that of Ki67 in all the experimental groups, with a Pearson correlation coefficient of 0.98. In addition, immunohistochemical expression of p53 in the hyperplastic squamous epithelium was very weak, in contrast to the more intense immunoreactivity observed in verrucous carcinoma and classic squamous cell carcinoma. There was an overlapping in the expression of p16, p21, and RGBP in all the experimental groups, being present in more than half the thickness of the epithelium in 50% to 100% cases in each study group. We therefore conclude that the pattern of Ki67 and p53 expression in verrucous carcinoma is readily reproducible and distinctly different from that observed in epithelial hyperplasia and that seen in invasive squamous cell carcinoma. Thus Ki67, and p53 immunostains are reliable adjuncts that may be helpful in resolving diagnostic problems associated with verrucous carcinoma. PMID- 15894932 TI - Expression of eukaryotic initiation factor 4E in astrocytic tumors. AB - Upregulation of expression of the eukaryotic initiation factor 4E (eIF4E) has been identified in breast carcinomas, squamous cell carcinomas in the head and neck regions, and transitional cell carcinomas of urinary bladder. In this immunohistochemical study, eIF4E protein expression was investigated in human brain tissue from patients without central nervous system diseases and brain biopsy tissues from patients with anaplastic astrocytoma and glioblastoma multiforme. Expression of eIF4E protein was observed in normal pyramidal neurons but not in neuroglial components. In anaplastic astrocytoma and glioblastoma multiforme, there was diffuse uniform expression of eIF4E immunoreactivity in malignant astrocytes. A similar pattern of immunoreactivity was also present in proliferative endothelial cells and vascular lining endothelial cells in glioblastoma multiforme. This study provides evidence that eIF4E is upregulated in high-grade astrocytic tumors. As in other malignancies, a high level of eIF4E may play an important role in the neoplastic transformation, angiogenesis, and tumor growth in astrocytic tumors. Because eIF4E is crucial in regulation of tumor growth, eIF4E could be a potential target for inhibitors as an adjuvant therapy for brain tumors. PMID- 15894933 TI - Expression of urokinase-type plasminogen activator receptor (uPAR) in primary central nervous system neoplasms. AB - The cellular receptor for urokinase-type plasminogen activator receptor (uPAR) is a member of the glycosylphosphatidylinositol (GPI) anchored protein family. It is a specific cell surface receptor for its ligand, urokinase-type plasminogen activator, which catalyzes the formation of plasmin from plasminogen to generate the proteolytic cascade and leads to the breakdown of the extracellular matrix. uPAR has been shown to correlate with a propensity to tumor invasion and metastasis in several types of non-central nervous system tumors. In this study, the authors examined the immunohistochemical expression of uPAR in 65 primary brain tumors (5 pilocytic astrocytomas, 5 diffuse astrocytomas, 6 anaplastic astrocytomas, 8 glioblastomas, 5 oligodendrogliomas, 4 oligoastrocytomas, 6 anaplastic oligoastrocytomas, 4 gangliogliomas, 4 ependymomas, 5 medulloblastomas, 6 schwannomas, 5 meningiomas, 2 atypical meningiomas). The specimens were evaluated for intensity of immunostaining (0-3 scale), cellular localization of staining, and specific or unique patterns of staining. Some degree of uPAR expression was observed in all tumors. A significant positive correlation (P = 0.0006) between tumor grade and staining intensity was identified within the astrocytoma/glioblastoma subgroup, suggesting a possible correlation with anaplastic change and propensity to tumor invasion. Expression of uPAR in nonmalignant, noninvasive tumors such as schwannoma and meningioma suggests that uPAR may have other biologic functions in addition to promotion of tumor invasion. PMID- 15894934 TI - Immunohistochemical Localization of Fas-associated phosphatase-1 (FAP-1) in Alzheimer disease hippocampus. AB - Fas-associated phosphatase-1 (FAP-1) is a regulatory peptide inhibiting apoptotic signal transduction via the death receptor Fas. Because apoptosis is a common mechanism leading to neuronal death in neurodegenerative disorders, the authors investigated the immunohistochemical distribution of FAP-1 in the hippocampus of elderly control subjects and Alzheimer disease (AD) patients. The current study provides the first evidence that FAP-1 is localized in the human hippocampus in pyramidal neurons of the hippocampal subfields CA1-4 and in granular cells. Cellular and extracellular FAP-1 intensity was increased in some control subjects and AD patients, but was not related to the stage of the illness. Rather, these data may indicate a general role for FAP-1 in neuronal death both in adult CNS and during the course of neurodegenerative disorders. PMID- 15894935 TI - HER2 positivity in breast carcinoma: a comparison of chromogenic in situ hybridization with fluorescence in situ hybridization in tissue microarrays, with targeted evaluation of intratumoral heterogeneity by in situ hybridization. AB - An accurate and reproducible assay method for determining HER2 status is crucial, as a positive HER2 gene status is an eligibility requirement for Herceptintrade mark therapy. Although immunohistochemical (IHC) assessment is both practical and inexpensive, a worrying trend of high false-positive rates has been reported. Fluorescence in situ hybridization (FISH) is the universally accepted gold standard for confirming IHC 2+ cases and ambiguous results but is costly and requires specialized equipment and technical expertise. Chromogenic in situ hybridization (CISH) amalgamates the practical advantages of IHC with the reproducibility of FISH, and high concordance between the CISH and FISH methods has been reported in conventional sections. Tissue microarrays (TMAs) allow high throughput of specimens, and HER2 status assessment in TMA cores using IHC and FISH has correlated well with scores in conventional sections. The authors used TMA technology to compare the efficacy of ZYMED(R) CISH with PathVysiontrade mark FISH in a cohort of 119 archival breast resection cases and investigated possible intratumoral heterogeneity in a "mini-array" of 21 HercepTest "equivocal"/2+ cases. Concordance between FISH and CISH in TMA sections was 99%. All prescored 2+ HercepTest cases were nonamplified. Four 3+ HercepTest cases were classed as potential false-positives. The authors suggest that confirmatory ISH should be performed on all positive HercepTest cases. CISH was easier to perform and quicker to enumerate than FISH. The authors conclude that CISH is a practical alternative to FISH as a confirmatory tool for HER2 gene amplification status. Intratumoral heterogeneity did not affect the patient's HER2 status. PMID- 15894936 TI - Pleomorphic atypical extraosseous ewing sarcoma in a 25-year-old woman: a cytogenetic diagnosis. PMID- 15894937 TI - Pain and anxiety control in Down syndrome. AB - The most recent pain and anxiety control techniques employed in patients with Down syndrome are described in relation to how cooperative the patient is and what assessment is made of his or her general condition. The growing deinstitutionalization of these patients and the growing social opportunities for them are matched by an increasing demand for dental treatment. Down syndrome is thus one of the disabilities with which the dentist is most likely to come into contact in his own surgery. Pain control is one of the keys to building the relationship between physician and patient and obtaining patient compliance in the conviction that only regular controls can maintain the oral health of these patients. A local anaesthetic, combined when possible with psychological techniques, remains the approach of choice and only a level of compliance that is really wanting or the need to carry out numerous operations in a single session should lead the dentist to choose conscious sedation or a general anaesthetic. The choice of technique, however, should take into account any systemic disease present, such as congenital heart and neurological diseases, bearing in mind that conscious or deep sedation and general anaesthesia should only be adopted in a hospital environment and require the presence of an anaesthesiologist. PMID- 15894938 TI - Overdentures on natural teeth: a new approach. AB - AIM: The study presents a new type of copings for overdentures on natural teeth. METHODS: A new type of custom-made copings was prepared on 10 extracted teeth and their marginal fit was observed microscopically by means of a mechanical device, and software was employed to measure the gap. RESULTS: The marginal fit evaluation gave satisfactory values with mean values of the gap measurements below the clinically accepted limits (mean gap: 25.3 microm; minimum 7.3 microm, maximum 56.5 microm). CONCLUSIONS: The advantages of these new copings are: the rapidity of their preparation; the protection of the root canal treatment, because the coping with this chair-side method is prepared and cemented in one session; the low costs. PMID- 15894939 TI - Comparison of masticatory dysfunction in temporomandibular disorders and fibromyalgia. AB - AIM: Many theories have been proposed as to the relationship between fibromyalgia (FM) and temporomandibular disorders (TMD). The aim of this study was to investigate the clinical features of the involvement of the stomatognathic system in patients with fibromyalgia, and to compare signs and symptoms of masticatory dysfunction between TMD and FM patients. METHODS: The study sample consisted of 30 subjects with fibromyalgia recruited at the Rheumatology Disease Department, University of Pisa, Italy, and of 30 with temporomandibular disorders at the Section of Prosthetic Dentistry, Department of Neurosciences, University of Pisa, Italy. Masticatory dysfunction has been compared between the 2 groups by means of a clinical assessment conducted according to the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD). Patients of the TMD group underwent a rheumatologic examination to investigate for the presence of previously undiagnosed fibromyalgia. RESULTS: As regards specific clinical variables and RDC/TMD diagnoses, differences between the 2 groups appear to be not relevant, with the exception of the values of mouth opening, both voluntary and passive, and the presence of trigger points. FM patients showed a decrease in both maximum voluntary and passive mouth opening which was about 2 mm worse than that recorded in TMD patients. FM patients also showed a significantly higher number of trigger points, even though muscle palpation evoked tenderness in the same number of sites in the 2 groups. CONCLUSIONS: This study showed that most patients with fibromyalgia (86.7%) report signs and symptoms localized at the stomatognathic system; by contrast, only a minority of patients with temporomandibular disorders (10%) are actually affected by fibromyalgia. PMID- 15894940 TI - 17-beta-estradiol and progesterone serum levels in temporomandibular disorder patients. AB - AIM: Considering the hypothesis that some temporomandibular joint (TMJ) tissues could be a potential target for sexual hormones, the aim of the study was to evaluate estrogen (17-beta-estradiol) and progesterone serum levels in a young adult population affected by articular forms of temporomandibular disorders (TMD) versus a control group of healthy subjects. METHODS: A total of 35 patients with Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) Axis I Group II diagnosis of disk displacement and/or Group III diagnosis of arthralgia, osteoarthritis or osteoarthrosis, were recruited at the Section of Prosthetic Dentistry, Department of Neurosciences, University of Pisa, Italy, along with a sex- and age-matched group of 24 healthy controls. In all patients, 17-beta estradiol, progesterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH) serum levels were determined using a radioimmunoassay. A T-test was performed to compare mean 17-beta-estradiol and progesterone serum levels in the TMD groups with mean serum levels of their respective control groups. Significance was set at p<0.05. RESULTS: Significant differences between patients affected by TMJ disorders and healthy controls were found for serum concentration of 17-beta-estradiol, both in males (p<001) and in the luteal phase of the menstrual cycle in females (p<0.05). No difference was found for progesterone serum levels in the different experimental samples. CONCLUSIONS: The results of this study suggest that high serum estrogens levels might be implicated in the physiopathology of temporomandibular joint disorders, since subjects with these pathologies showed significantly higher serum levels with respect to a group of healthy controls. PMID- 15894941 TI - Temporomandibular disorders and sleep disorders: which relationship? AB - AIM: The aim of this case-control study was to establish statistically significant associations between temporomandibular disorders (TMD) and obstructive sleep apnea (OSA) or excessive daytime sleepiness (EDS) using, respectively, the Douglass Sleep Disorders Questionnaire and the Epworth Sleepiness Scale. METHODS: A total of 100 patients affected by TMD and 100 age- and sex-matched controls randomly chosen from the general population, answered the 2 questionnaires, which investigated their sleep habits, sleep hygiene and sleep disorders. OSA was considered as present if the total Douglass questionnaire score reached or exceeded the cut-off point of 32 for women and 36 for men. The total score on the Epworth Sleepiness Scale was calculated for each subject, a total >10 considered to indicate the presence of EDS. The data were analysed by using the chi squared test. RESULTS: EDS was found in 19% of the patients and in 10% of the control subjects, and OSA in 6 patients and in 4 control subjects. No statistically significant association emerged between TMD and OSA. EDS was more frequent in patients with myofascial pain. On the basis of the results of the first questionnaire, we were able to develop a profile of OSA patients: they are predominantly smokers, middle-aged (40-60 years old), overweight men. CONCLUSIONS: Although most TMD patients complain of poor sleep quality, the age and sex prevalence of OSA was lower than that of TMD, and EDS was found to be present in only a few cases. PMID- 15894942 TI - Early childhood caries. Oral health survey in kindergartens of the 19th district in Rome. AB - AIM: The aim of this cross-sectional survey was to determine the prevalence of early childhood caries (ECC) in a pediatric sample randomly chosen in public kindergartens or the 19th district in Rome, according to the WHO guidelines and to access the dental and orthodontic status of the affected children. METHODS: A total of 546 children aged 3-to 5-years were examined. Clinical examination had been executed from January 1999 to June 2000 by 4 calibrated operators; WHO clinical assessment form and anamnestic questionnaires for parents were performed. RESULTS: Children affected by ECC with a dmft score of 6.3 were 8.2% and 93% were ''treatment need'' (TN); low levels of oral health was significantly related with father's low socioeconomical status, both occupational and educational: most of these families were from less industrialized countries; 40% of these kids used to suck sucking-bottles with honey: many others used to drink too much sweet things (milk and sugar, fruit juice etc.); the most frequent orthodontic features in the ECC group were: brachifacial (26.7%), convex profile (70%), 3rd class terminal plane (30%). CONCLUSIONS: The results obtained point out that information, health education and health promotion are lacking throughout this part of roman population. Primary preventive dentistry programs should be therefore organized targeting parents, nursery schools and kindergartens' operators and pediatricians. PMID- 15894943 TI - Orthognatic therapy for the resolution of a presumed mandibular follicular cyst. Clinical case. AB - The case of a presumed mandibular follicular cyst associated with the retention of 4.5 in a 12 year-old patient is described. Objective examination revealed the absence of 4.5 in the dental arch. Orthopantomography revealed a monofocal area of radiotransparency with a sharp outline by 4.5 tooth. The lesion was treated only by fixed multiband orthodontic therapy to reopen the space in the dental arch. This treatment permitted the total remission of the cystic lesion without any surgical action, as confirmed by the following radiographical exams. PMID- 15894944 TI - Benign cementoblastoma: a clinical case of conservative surgical treatment of the involved tooth. AB - The benign cementoblastoma (BC) or ''true'' cementoma is a rare benign neoplasm arising from the odontogenic ectomesenchyme and representing about 1% to 6.2% of all odontogenic tumors. The BC more frequently affects young males in an age range of 20-30 years, occurring in the mandible about 3 times more than in the maxilla, and it is always physically attached to the tooth roots. This tumor is often asymptomatic until it produces pain, expansion or swelling of the jaw segment or compression of the inferior alveolar nerve. Early diagnosis is essential to save the tooth by enucleating the tumor, filling the root canals and apicectomy or curettage of the affected roots. A case of BC embedding the mandibular first right molar and resorbing the vestibular cortical bone, in a 48 year-old male, is reported. The radiographic examination showed a well-defined mixed-density unilocular mass, confluent with both the tooth roots of the mandibular first molar and surrounded by a radiolucent rim. A combined endodontic surgical treatment was performed with the aim to remove the tumor while saving the tooth. Histological findings, differential diagnosis and surgical treatment of the tumor are discussed and compared with similar cases in the literature. PMID- 15894945 TI - Residual and inflammatory radicular cysts. Clinical and pathological aspects of 2 cases. AB - Inflammatory odontogenic cysts include radicular cysts and its etiological variance, residual cysts. Among these lesions, the radicular cyst is the most frequent. It is caused by the growth of remnants of Malassez cells involved in the development of the dental organ. Clinically, radicular cysts are difficult to diagnose. Histologic diagnosis is of primary importance in order to definitely discriminate the different kinds of periapical lesions. In this paper, the clinical, radiographic, etio-physio-pathological and microscopic features of these pathological formations are described. A case of a large radicular cyst and a case of residual cyst are reported and the surgical treatment and histologic differential diagnosis are presented. PMID- 15894946 TI - Effects of angiotensin-converting enzyme inhibitor plus irbesartan on maximal and submaximal exercise capacity and neurohumoral activation in patients with congestive heart failure. AB - BACKGROUND: In patients with symptomatic congestive heart failure receiving optimal therapy with an angiotensin-converting enzyme (ACE) inhibitor and a beta blocker, the impact of using an angiotensin receptor blocker on submaximal exercise capacity and on neurohumoral activation at rest and during stress has not been investigated. METHODS: Thirty-three patients with congestive heart failure, New York Heart Association II or III symptoms, and left ventricular ejection fraction 25.5% +/- 7.2% treated with an ACE inhibitor and a beta-blocker were recruited. Patients were randomly assigned to receive irbesartan 150 mg per day (n = 22) or a placebo (n = 11) for 6 months. Maximal exercise capacity was assessed using a ramp protocol. Submaximal exercise duration was assessed using a constant load protocol, and plasma norepinephrine and angiotensin II (A-II) were measured in resting state, at 6 minutes, and at peak exercise. RESULTS: Patients treated with irbesartan presented a 26% increase in submaximal exercise time (+281 seconds, P = .08) whereas exercise duration increased by only 7% in patients treated with a placebo (+128 seconds, P = NS irbesartan vs placebo). Norepinephrine levels increased to a similar extent in both groups, whereas A-II levels did not increase or change in response to therapy. CONCLUSIONS: Dual A-II suppression with an ACE inhibitor plus irbesartan provides a small but a significant increase in submaximal exercise capacity. This beneficial effect is observed despite no significant changes in maximal exercise capacity, and in resting or exercise-induced increase in neurohumoral activation. PMID- 15894947 TI - An alternative approach for exercise prescription and efficacy testing in patients with chronic heart failure: a randomized controlled training study. AB - BACKGROUND: Prescription of endurance training and documentation of its efficacy in patients with chronic heart failure (CHF) is usually done with reference to maximal ergometric measurements which are subject to form on the day changes and motivational influences. However, the use of submaximal parameters might represent an alternative approach. METHODS: This was tested in 54 patients with CHF (57 +/- 10 years, NYHA II: n = 40; NYHA III: n = 14) who were randomized into training (T; n = 26) or control group (CO; n = 28). Training consisted of 45 minutes cycling at an intensity corresponding to the anaerobic threshold (AT) and was conducted for 12 weeks, 4 to 5 times per week. Cardiorespiratory exercise testing was done before and after the experimental phase. Changes in well-being were investigated using a 5-point Likert scale. RESULTS: A significant rightward shift in the heart rate curve was demonstrated in T compared with CO (P = .01; T: decrease in resting heart rate by 8 per minute, during exercise by 7 to 11/min; CO: -1 and -1 to -3 per minute, respectively). Anaerobic threshold increased significantly by 11.6% in T (+0.11 +/- 0.11 L min(-1) oxygen uptake) compared with CO (-0.02 +/- 0.10 L min(-1)). Positive changes in well-being were significantly larger in T (P < .01). CONCLUSIONS: In patients with CHF, training can be prescribed and its efficacy can be evaluated by the exclusive use of submaximal parameters. Anaerobic threshold represents an appropriate training intensity in this population. PMID- 15894948 TI - Thrombolytic therapy for eligible elderly patients with acute myocardial infarction. PMID- 15894950 TI - Assessment of left ventricular mass using echocardiography: use of fundamental imaging. PMID- 15894952 TI - Improvements in quality improvement. PMID- 15894953 TI - A critical appraisal of current models of risk stratification for percutaneous coronary interventions. PMID- 15894954 TI - The Women's Recovery from Sternotomy (WREST) study: design of a randomized trial of a novel undergarment for early use after sternotomy. AB - BACKGROUND: Clinicians who work with women poststernotomy often suggest that they wear a supportive brassiere to ameliorate pain, discomfort, and potential wound complications. There is no empirical evidence that supports this practice. METHODS: Despite methodological challenges, a clinical trial is currently underway to investigate the efficacy of early use of a novel undergarment after sternotomy. Women (N = 430) having first time sternotomy in 9 Canadian centers will be randomized to receive either the usual care of the institution or early use of a novel undergarment. Follow-up is planned over 12 postoperative weeks. Coprimary outcomes are pain, discomfort, and return to function. Wound healing is a secondary outcome. An economic evaluation substudy is also underway. CONCLUSIONS: The WREST Study is a unique ongoing trial examining the efficacy of a novel undergarment in reducing women's pain and enhancing their comfort and return to function. The findings of the trial and its economic substudy will enable health care providers to make rational evidence-based clinical decisions regarding women's early care after sternotomy. PMID- 15894956 TI - Systolic hypertension: an overview. PMID- 15894957 TI - Evaluation and management of patients with uncontrolled systolic hypertension: is another new paradigm really needed? PMID- 15894958 TI - Barriers to hypertension control. PMID- 15894959 TI - Improving blood pressure control by tailored feedback to patients and clinicians. PMID- 15894960 TI - Diet and blood pressure: applying the evidence to clinical practice. PMID- 15894961 TI - Impact of invasive management versus noninvasive management on functional status and quality of life following non-Q-wave myocardial infarction: a randomized clinical trial. AB - BACKGROUND: Multiple studies have examined whether clinical outcomes are improved by invasive management following non-Q-wave myocardial infarction (NQWMI). However, it remains unclear whether functional status and quality of life are affected by an invasive strategy. METHODS: Following NQWMI, we randomized 88 patients to invasive management vs noninvasive management. The primary end point was functional status assessed at 12 months using maximal endurance exercise treadmill testing measured in metabolic equivalents. Secondary end points included changes in scores between baseline and 12 months on the Duke Activity Status Index, the Seattle Angina Questionnaire, and the Medical Outcomes Study 36 Item Short-Form Survey. RESULTS: Of the 42 patients in the invasive arm, 83% underwent initial angiography. Of the 46 patients in the noninvasive arm, 91% underwent initial stress testing. Inhospital and 12-month revascularization rates were similar in the 2 arms (24% vs 22%, P > or = .99; 31% vs 30%, P > or = .99). Maximal endurance exercise treadmill testing was also similar at 12 months (7.8 vs 6.7 metabolic equivalents, P = .24). Patients in the invasive arm showed improved functional status by mean difference in their Duke Activity Status Index scores (4.3 vs -3.5, P = .04). Improvements in angina-specific quality of life for patients in the invasive arm were demonstrated by the Seattle Angina Questionnaire measures of anginal stability (21.6 vs -5.3, P = .02), anginal frequency (22.9 vs 2.3, P = .02), treatment satisfaction (11.2 vs -10.3, P = .02), and disease perception (24.7% vs 10.9%, P = .07). CONCLUSIONS: Compared with patients undergoing noninvasive management of NQWMI, patients undergoing invasive management have some measures indicative of improved functional status. PMID- 15894962 TI - Potential antiatherogenic and anti-inflammatory properties of sevelamer in maintenance hemodialysis patients. AB - BACKGROUND: Patients affected by end-stage renal disease (ESRD) demonstrate a very high cardiovascular risk mediated by traditional cardiovascular risk factors as well as abnormal mineral metabolism and a state of chronic inflammation. Sevelamer is a nonabsorbable non-calcium-based hydrogel with potential antiatherosclerotic properties. METHOD AND RESULTS: One hundred eight patients undergoing maintenance hemodialysis were randomized to sevelamer or calcium acetate as treatment for hyperphosphatemia. A coronary artery calcium score, as a measure of plaque burden, was calculated at baseline and 1 year, along with serial measurements of serum lipoproteins, beta2-microglobulin, and high sensitivity C-reactive protein (hs-CRP). At 1 year, coronary artery calcium score progressed significantly from baseline in calcium acetate-treated subjects ( P < .001) but not in sevelamer-treated patients (P = NS). Total cholesterol (P < .0001), low-density lipoprotein cholesterol (P < .0001), apolipoprotein B (P < .0001), beta2-microglobulin (P = .018), and hs-CRP (P < .002) decreased, and high density lipoprotein increased significantly (P = .036) from baseline in the sevelamer-treated subjects but not in subjects treated with calcium acetate despite the more frequent use of statins in the latter group (46% vs 22%, P < .05). The changes in total and low-density lipoprotein cholesterol, apolipoprotein B, and hs-CRP were significantly different between treatment groups (all P < .01). CONCLUSIONS: Sevelamer leads to favorable changes in lipids and inflammatory markers with potentially useful antiatherogenic effects in hemodialysis patients. PMID- 15894963 TI - Impact of the application of the American College of Emergency Physicians recommendations for the admission of patients with syncope on a retrospectively studied population presenting to the emergency department. AB - BACKGROUND: The goal of our study was to determine the accuracy of the American College of Emergency Physicians (ACEP) clinical policy in identifying patients with a cardiac cause for their syncope and its potential effect on syncope management. METHODS: Adult patients with syncope presenting to the emergency department (ED) from January 1996 to December 1998 were identified. Diagnosis was established retrospectively by reviewing medical records. The ACEP guidelines were applied to this population. RESULTS: Of the 200 patients identified, 115 (57.5%, 95% CI 60-64) were admitted from the ED and 24 (12%) were found to have cardiogenic syncope. Of the 24 patients with cardiac syncope, 23 were admitted. By applying ACEP level B recommendations to our population, all patients who on further workup were found to have cardiac syncope would have been admitted from the ED (100% sensitivity, 95% CI 86-100) and 81% of patients with no cardiac syncope would have been discharged from the ED (81% specificity, 95% CI 75-87). The admission rate would have been 28.5% (95% CI 22-35). By extending admission to patients satisfying level C in addition to level B recommendations, the sensitivity, specificity, and admission rate would have been 100% (95% CI 86 100), 33% (95% CI 26-40), and 71.0% (95% CI 64-77), respectively. CONCLUSION: High sensitivity and specificity in identifying patients with cardiogenic syncope and significant reduction in the hospital admission rate were observed by applying ACEP level B recommendations to patients presenting to our ED. Application of level C recommendations did not offer any advantage. PMID- 15894964 TI - Cigarette smoking is associated with increased circulating proinflammatory and procoagulant markers in patients with chronic coronary artery disease: effects of aspirin treatment. AB - BACKGROUND: Smoking is associated with endothelial dysfunction. Cytokines released by injured endothelium promote vascular interactions with leukocytes and platelets. We investigated whether (a) cigarette smoking is linked to increased cytokine production, which may mediate platelet activation and thrombin generation in chronic coronary artery disease (CAD), and (b) aspirin treatment inhibits smoking-related changes on cytokines, platelets, and thrombin. METHODS AND RESULTS: Plasma macrophage-colony-stimulating factor (M-CSF) and C-reactive protein (CRP) were measured in 100 patients with chronic CAD, 60 of whom were chronic smokers. Prothrombin fragments 1+2 and urinary 11-dehydro-thromboxane B2 (TXB2) were additionally measured in 60 of 100 patients (30 of whom were smokers) and in 24 healthy controls. Smokers (n = 20) matched for age, myocardial ischemia, and other risk factors with 20 nonsmokers entered a double-blind crossover trial of aspirin (300 mg/d for 3 weeks) versus placebo. Blood and urine measurements were repeated after each treatment. Compared with nonsmokers, smokers had 3-fold median M-CSF (1499 vs 476 pg/mL), 2-fold CRP (1.5 vs 0.8 mg/L), and higher 11-dehydro-TXB 2 (3.6 vs 2.1 ng/mg creatinine, P < .01 for all comparisons). After aspirin treatment, M-CSF, CRP, 11-dehydro-TXB 2 , and prothrombin fragments 1+2 remained higher in smokers compared with nonsmokers despite a significant reduction of these markers by aspirin (P < .05). M-CSF remained related to 11-dehydro-TXB 2 excretion during both treatment phases (P < .01) suggesting that cytokine-mediated thromboxane A 2 production was not altered by aspirin. CONCLUSIONS: Smoking is associated with increased M-CSF, CRP, and platelet activity. Although aspirin treatment reduces the proinflammatory and procoagulant markers in smokers, it does not abolish the proinflammatory effects of smoking in patients with chronic CAD. PMID- 15894965 TI - Microalbuminuria: a strong predictor of 3-year adverse prognosis in nondiabetic patients with acute myocardial infarction. AB - BACKGROUND: The aim of this study is to evaluate the significance of microalbuminuria (MA) as a 3-year prognostic index in nondiabetic patients with acute myocardial infarction (AMI). METHODS: One hundred seventy-five patients with AMI were followed prospectively for 3 years. The study end point was cardiac death or rehospitalization for an acute coronary event. RESULTS: Forty-two patients (24%) developed a new cardiac event during the follow-up. Microalbuminuria (P < .001), pulmonary edema during initial hospitalization (P < .001) and postinfarction angina (P = .0364), advanced age (P = .001), severe atherosclerosis (high Gensini score) (P = .036), ejection fraction <50% (P = .0013), history of bypass surgery (P = .0265), and early conservative management (P = .0214) were all associated with adverse prognosis. Cox proportional hazards regression analysis showed that MA was an independent predictor of 3-year adverse prognosis in all the models tested, with an adjusted relative risk for the development of a cardiac event ranging from 2.1 to 4.3. CONCLUSIONS: In nondiabetic patients with AMI, MA is a strong and independent predictor of an adverse cardiac event within the next 3 years. PMID- 15894966 TI - Correlation between the TIMI risk score and high-risk angiographic findings in non-ST-elevation acute coronary syndromes: observations from the Platelet Receptor Inhibition in Ischemic Syndrome Management in Patients Limited by Unstable Signs and Symptoms (PRISM-PLUS) trial. AB - BACKGROUND: The TIMI risk score for unstable angina and non-ST elevation myocardial infarction is an effective tool for predicting the risk of death and ischemic events among patients with non-ST elevation acute coronary syndromes, as well as for identifying those who are likely to benefit most from low-molecular weight heparin and glycoprotein IIb/IIIa inhibition. METHODS: To explore the pathobiologic basis for this interaction, we evaluated the relationship between the risk score, assessed at presentation, and angiographic findings among patients with non-ST elevation acute coronary syndromes. Angiographic data regarding thrombus, epicardial flow, and lesion severity were available for 1491 patients from the angiographic substudy of PRISM-PLUS. RESULTS: Patients with risk scores of 5 to 7 (N = 435) were more likely to have a severe culprit stenosis (81% vs 58%, P < .001) and multivessel disease (80% vs 43%, P < .001) compared to those with scores of 0 to 2 (N = 220). The probability of left main disease (P = .01), visible thrombus, and impaired flow in the culprit lesion also increased progressively with rising risk scores (P < .001). Of the risk indicators that comprise the score, history of coronary disease, advanced age, and ST changes showed the strongest association with severe epicardial disease. Positive biomarkers of necrosis, ST changes, and prior aspirin use emerged as stronger correlates of visible thrombus and/or impaired culprit artery flow. CONCLUSIONS: The TIMI risk score identifies patients who are more likely to have intracoronary thrombus, impaired flow, and increased burden of coronary atherosclerosis. These findings likely explain in part the particular benefit of potent antithrombin and antiplatelet agents among patients with higher risk scores. PMID- 15894967 TI - Depression and heart failure in patients with a new myocardial infarction. AB - BACKGROUND: Heart failure (HF) is a disabling chronic illness that is increasing in prevalence. Despite advances in its medical treatment, little is known about its psychosocial correlates. This investigation compared the prevalence of depression in patients with and without HF who were hospitalized with myocardial infarction (MI) and also at high psychosocial risk. METHODS: Baseline data from the 2444 participants in the ENRICHD clinical trial were used. Within 28 days of the MI, all patients were assessed for depression using Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria, for HF based upon left ventricular dysfunction or history of HF and for demographic and medical confounders. RESULTS: Eight hundred forty-seven patients (34.7%) met the criteria for HF. Major depression was observed in 43% in the HF group compared with 36% in the non-HF group (P < .001). Multivariate modeling showed a 38% increase in odds of HF for patients with major depression, after adjustment for sociodemographic factors and medical comorbidities (adjusted OR 1.38, 95% CI 1.09 1.76). CONCLUSIONS: These findings suggest that in patients with new MI who are at psychosocial risk, major depression is more prevalent in those who also have HF. Because major depression undermines adherence and lowers quality of life, special efforts to diagnose and treat it in post-MI patients with HF appear warranted. PMID- 15894968 TI - Data feedback and clinical process improvement in acute myocardial infarction. AB - BACKGROUND: Numerous reports have shown substantial undertreatment of acute myocardial infarction (AMI). The goal of this intervention study was to improve both the acute and discharge care of patients with AMI. Thirty-eight Tenet hospitals participated in a cardiac improvement collaborative called the Partnership for Change. Interventions included the implementation of a computerized concurrent data feedback system and the organization, staffing, and deployment of performance improvement teams at each hospital. METHODS: Data were collected on 11 394 patients with AMI between January 1, 2001, and June 30, 2002. We calculated rates of treatments including aspirin (first 24 hours and at discharge), beta-blocking agent (first 24 hours and at discharge), reperfusion using thrombolysis (first 30 minutes) or percutaneous intervention (first 2 hours), angiotensin-converting enzyme inhibitor (at discharge), smoking cessation counseling (during hospital stay), and referral to a cardiac rehabilitation program (at discharge). RESULTS: Statistically significant increases were seen for all indicators. Aspirin use during the first 24 hours increased from 86.4% to 96.5% and use at discharge increased from 70.0% to 87.4%. The use of beta blocking agents during the first 24 hours increased from 51.4% to 88.4% and use at discharge increased from 62.4% to 83.5%. Reperfusion during the first 2 hours increased from 25.9% to 35.3%. Discharge use of angiotensin-converting enzyme inhibitors increased from 29.3% to 43.3%. Smoking cessation counseling increased from 35.1% to 80.6%. Referral to a cardiac rehabilitation program increased from 16.9% to 41.7%. All P values were <.001. CONCLUSIONS: The implementation of a rapid-cycle computerized concurrent data feedback system along with on-site process improvement teams was associated with a substantial increase in the use of both acute and discharge treatments for AMI. PMID- 15894969 TI - Impact of program duration and contact frequency on efficacy and cost of cardiac rehabilitation: results of a randomized trial. AB - BACKGROUND: Secondary prevention through cardiac rehabilitation (CR) has been recommended for most patients with coronary artery disease (CAD). Although generally reimbursed for 3 months, to date, optimal CR program duration and frequency of patient contact has yet to be identified. This study compared standard (33 sessions for 3 months) versus distributed (33 sessions for 12 months) CR for effects on exercise variables, risk factors, health-related quality of life (HRQL), depressive symptoms, and direct costs to the cardiac health care system. METHODS: We randomly assigned 392 patients to either standard CR (n = 196) or distributed CR (n = 196). Outcomes were cardiorespiratory fitness, daily physical activity, coronary risk factors, generic and heart disease HRQL, and depressive symptoms, measured 12 and 24 months after program intake. Secondary outcomes included these variables measured after 3 months. Costs to the cardiac health care system were determined 2 years after program initiation. RESULTS: Both groups showed improvements over time in cardiorespiratory fitness, daily physical activity, low-density lipoprotein cholesterol, generic and heart disease HRQL, and depressive symptoms. Over time, blood pressure and body mass index values worsened. Smoking status, high-density lipoprotein cholesterol, and triglyceride levels remained unchanged. There were no clinically meaningful or statistically significant between group differences for outcomes at 12 or 24 months. The costs of the programs to the cardiac health care system were not different. CONCLUSIONS: From a clinical standpoint, this study indicates that both standard and distributed program formats serve patients with CAD equally well over the longer term. Programs could use either program delivery model (standard or distributed) depending on patient or program needs. Costs to the cardiac health care system are similar. PMID- 15894970 TI - The relation of renal function to ischemic and bleeding outcomes with 2 different glycoprotein IIb/IIIa inhibitors: the do Tirofiban and ReoPro Give Similar Efficacy Outcome (TARGET) trial. AB - BACKGROUND: Renal function significantly impacts morbidity and mortality after a percutaneous coronary intervention. Platelet glycoprotein (GP) IIb/IIIa inhibitors reduce ischemic complications during percutaneous coronary intervention; little is known of whether their safety and efficacy are influenced by renal function. In particular, whether outcome differences exist between agents that are renally excreted (tirofiban) or not (abciximab) in patients with mild renal impairment is not known. METHODS: The TARGET trial randomized 4623 patients to tirofiban or abciximab. In this analysis, patients were grouped in creatinine clearance quartiles (<70, 70-90, 90-114, >114 mL/min) and analyzed for efficacy and bleeding risk. Univariate and multivariate analyses were performed to identify interactions between GP IIb/IIIa inhibitor used and creatinine clearance with respect to ischemic outcomes and bleeding. RESULTS: Using unadjusted logistic regression tests for trend, 30-day death/myocardial infarction/urgent target vessel revascularization was greater in patients with lower creatinine clearances (7.3%, 8.5%, 5.1%, and 5.8%, P = .005), as were both major and minor bleeding. There was no interaction between assigned GP IIb/IIIa inhibitor, creatinine clearance and ischemic outcome, major bleeding or minor bleeding. CONCLUSIONS: Both ischemic and bleeding complications are highest in the lowest creatinine clearance quartile of patients treated with GP IIb/IIIa inhibitors. Although tirofiban is renally cleared and abciximab is not, there was no interaction between these GP IIb/IIIa inhibitors and creatinine clearance regarding ischemic or bleeding events. PMID- 15894971 TI - Stent implantation, but not pathogen burden, is associated with plasma C-reactive protein and interleukin-6 levels after percutaneous coronary intervention in patients with stable angina pectoris. AB - BACKGROUND: The systemic inflammatory response to percutaneous coronary intervention (PCI) is associated with recurrent cardiac events; however, the pathophysiology of this inflammatory response is not well understood. The present study was performed to investigate the role of pathogen burden of infection in determining the magnitude of C-reactive protein (CRP) and interleukin 6 (IL-6) response to PCI. METHODS: One hundred patients with stable angina pectoris undergoing elective PCI at a single center were recruited. Antibodies against cytomegalovirus, Chlamydia pneumoniae , Epstein-Barr virus, Helicobacter pylori , and herpes simplex virus types 1 and 2 were determined before PCI. Plasma CRP and IL-6 levels were measured before and 6, 24, 48, 72 hours after PCI and data presented as area under the curve. RESULTS: Plasma CRP and IL-6 concentrations increased significantly after PCI. Neither antibodies against single nor multiple pathogens were associated with the CRP or IL-6 response to PCI. No correlations were found between the inflammatory markers and troponin T levels after PCI. With the exception for CRP and body mass index (R = 0.20, P < .05), neither risk factors for coronary heart disease nor medication but stent implantation was associated with increased plasma CRP (76 vs 61 mg/L, P < .005) and IL-6 (74 vs 64 pg/mL, P < .005) levels after PCI. CONCLUSION: Stent implantation, but not pathogen burden, is associated with the plasma CRP and IL-6 response to PCI. PMID- 15894972 TI - Renal and iliac artery stenting by interventional cardiologists and vascular surgeons: the Foundation to Advance Medical Education (FAME) initiative. AB - The American College of Cardiology, in concert with the Society for Coronary Angiography and Interventions and the Society for Vascular Surgery, planned and implemented an initiative to teach stenting of renal and iliac arteries to their peers in practice. Supported by the Foundation to advance medical Education (FAME), the initiative involved 25 preceptees and 6 preceptors and included a didactic portion (a remote learning exercise), an animal practicum, a procedural simulation, and a preceptor/preceptee training segment, all of which was followed by evaluations by the participants. Assessments of the success of the initiative differed between preceptees, who were positive about the experience, and preceptors, who were more critical of the endeavor (with surgeons more critical than cardiologists). Technical problems, such as obtaining temporary licensure to practice medicine in another state, interfered with the hands-on experiences of several preceptees. Programs such as the FAME initiative will grow in importance as new technologies continue to be introduced into medical practice and the need to train physicians in practice in their use expands. PMID- 15894973 TI - Marked differences between patients referred for stress echocardiography and myocardial perfusion imaging studies. AB - BACKGROUND: Patients referred for stress echocardiography may differ significantly from those referred for stress myocardial perfusion imaging (MPI). Such differences, if present, should be considered when comparing the accuracy and discriminatory performance of these tests. METHODS: We prospectively collected demographic and clinical information on all stress imaging studies performed at our institution between 1998 and 2001. The data were reviewed, summarized, and compared using the t test and chi2 test where appropriate. RESULTS: Of 5320 stress imaging studies performed, 3383 were stress echocardiographies and 1937 were MPI studies. Patients referred for MPI were older (59 vs 54, P < .0001), and more likely to have diabetes (32% vs 20%, P < .0001), prior myocardial infarction (39% vs 15%, P < .0001), and prior revascularization (38% vs 12%, P < .0001). Pharmacologic stress testing was much more common in the MPI group (66% vs 17%, P < .0001). More patients referred for MPI had decreased left ventricular function (23% vs 7%, P < .0001) and abnormal stress test results (41% vs 18%, P < .0001). CONCLUSIONS: Patients with a history of myocardial infarction, revascularization, or higher risk profiles are more likely to be referred for MPI compared to stress echocardiography at our institution. These differences in referral patterns are likely to exist in other centers, and it is reasonable to assume that systematic differences in test selection occur, resulting in patient populations with differing clinical risk profiles. Caution in interpreting analyses comparing the accuracy of stress imaging modalities is appropriate. PMID- 15894974 TI - Assessment of patients with low-risk chest pain in the emergency department: Head to-head comparison of exercise stress echocardiography and exercise myocardial SPECT. AB - OBJECTIVES: The aim of the study was to compare head-to-head the performance of exercise tolerance test-stress echocardiography (ex-Echo) and exercise stress perfusion nuclear imaging (exercise-single-photon emission computed tomography [ex-SPECT]) for the diagnosis of coronary artery disease (CAD) in patients evaluated at the chest pain unit with delay from chest pain (CP) onset. BACKGROUND: As an early triage strategy for CAD in emergency medicine, ex-Echo could have the advantage of widespread availability and low costs. METHODS: In the years 2000-2002, 503 consecutive patients (mean age 60 years) with recent (<24 hours) CP and nonischemic electrocardiogram (ECG), in whom CAD remained undiagnosed after first line 6-hour work-up including serum markers of myocardial injury and resting echocardiogram, underwent ex-Echo and ex-SPECT within 24 hours. Patients with (+)ex-Echo or (+)ex-SPECT or (+)ex-ECG or abnormal troponin I were referred to coronary angiography; otherwise, they were discharged and followed up. End points were coronary stenosis > or =50% and cardiovascular events at 6-month follow-up. RESULTS: Ninety-nine patients (20%) had (+)ex-Echo and 121 (24%) (+)ex-SPECT; CAD was diagnosed in 81% and 67%, respectively; positive tests were concordant in 69%. In negative ex-Echo and ex-SPECT, final evidence of CAD emerged in 14 and 13, respectively. Ex-Echo demonstrated higher accuracy than ex-SPECT (93% +/- 1% vs 89% +/- 1%), optimal specificity (95% +/- 5% vs 90% +/- 5%), and positive predictive value (81% +/- 4% vs 67% +/- 4%); moreover, in the case of (-)ex-ECG, observed effective likelihood ratio indicates a (+)synergy between ex-ECG and ex-Echo. CONCLUSIONS: Ex-Echo can be an effective diagnostic strategy in the early triage of CP patients, improving diagnosis in case of (-)ex-ECG and reducing unnecessary angiography number. Its drawback is represented by the 5% of missed diagnosis. PMID- 15894975 TI - A Doppler-derived index of combined left ventricular systolic and diastolic function is an independent predictor of cardiovascular mortality in elderly men. AB - BACKGROUND: Few studies on echocardiographic and Doppler indices of cardiac function as predictors for cardiovascular mortality have been performed in the general population. The novel myocardial performance index (MPI, also denoted as Tei-Doppler index) has been suggested to reflect both left ventricular (LV) systolic and diastolic function. METHODS: We investigated whether 10 different echocardiographic and Doppler indices of cardiac function predicted cardiovascular mortality by using a population-based cohort of 583 seventy-year old men with a mean follow-up time of 6.8 years. RESULTS: In a multivariate Cox proportional hazard analysis for a 1-SD increase including LV ejection fraction, E/A ratio, MPI, and traditional cardiovascular risk factors (previous myocardial infarction, hyperlipidemia, hypertension, diabetes, smoking status, and LV hypertrophy), both ejection fraction (hazard ratio [HR] = 0.59, 95% CI 0.38-0.90, P < .05) and MPI (HR = 2.92, 95% CI 1.71-4.97, P < .0001) were found to be independent predictors for cardiovascular mortality. After excluding subjects with coronary heart disease, only MPI remained a significant predictor (HR = 2.18, 95% CI 1.11-4.27, P < .03, for 1-SD increase), and neither ejection fraction, E/A ratio, nor any of the cardiovascular risk factors remained significant predictors. CONCLUSION: Myocardial performance index is a predictor for cardiovascular mortality, independent of other measurements of cardiac function and of traditional cardiovascular risk factors in elderly men free from coronary heart disease. PMID- 15894976 TI - Left ventricular outflow tract obstruction provoked during dobutamine stress echocardiography predicts future chest pain, syncope, and near syncope. AB - BACKGROUND: Although dobutamine stress echocardiography (DSE) is associated with dynamic left ventricular (LV) obstruction, it is unknown whether such obstructive event, in general, and the specific site of obstruction, in particular, have unique clinical prognostic significance. We sought to determine whether dynamic LV outflow tract (LVOT) versus LV midcavitary obstruction provoked during DSE would predict future chest pain, syncope, and/or near syncope. METHODS: Two hundred thirty-seven patients (145 men and 92 women, mean age 58 +/- 13 [+/-SD] years) without DSE-provoked ischemia underwent continuous wave Doppler interrogation to detect any inducible dynamic flow obstruction. Patients were prospectively followed for a mean duration of 31 +/- 13 months. RESULTS: One hundred fifty-four of 237 patients had no provoked LV obstruction (group 1). Fifty-four (22.8%) had provoked LV midcavitary (group 2) obstruction, and 29 (12.2%) had outflow tract (group 3) obstruction. During follow-up, chest pain occurred more frequently in groups 2 (46%, P < .05) and 3 (52%, P = .05) as compared with group 1 (31%). A higher incidence of syncope and/or near syncope was noted in group 3 (21% vs 9% in group 1). LVOT obstruction but not midcavitary obstruction was a significant predictor of future chest pain (relative risk 2.63, P = .0021) and syncope and/or near syncope (relative risk 3.11, P = .036). Kaplan Meier analysis showed a significantly less event-free survival (P = .025) for the combined end point of chest pain, syncope, and/or near syncope in patients with LVOT obstruction. CONCLUSIONS: This is the first prospective study to identify the differential prognostic implications of the site of dynamic obstruction noted during DSE. Our results demonstrate that DSE-provoked LVOT obstruction is an independent positive predictor of future episodes of chest pain and syncope and/or near syncope. These findings warrant larger studies addressing treatment options to ameliorate symptoms in this subgroup of patients. PMID- 15894977 TI - Effect of coronary artery bypass grafting and aneurysmectomy on QT dispersion in moderate or severe left ventricular dysfunction. AB - BACKGROUND: The aim of the study was to evaluate the effect that coronary artery bypass grafting (CABG) and aneurysmectomy in the same session have on QT dispersion (QTd) in moderate or severe left ventricular dysfunction. METHODS: Forty-four patients underwent QT interval analyses before and 1 month after aneurysmectomy and CABG. QT interval and QTd were corrected for heart rate using the square root formula of Bazett (QTCB), the cubic root formula of Fridericia (QTCF), and the linear formula of Sagie et al (QT CS ). RESULTS: The mean pre- and postsurgery QTd results were 65.29 +/- 29.25 and 51.76 +/- 18.49 milliseconds, respectively; the corresponding findings for QT CF were 68.06 +/- 31.26 and 55.16 +/- 24.56 milliseconds; and the corresponding findings for QT CS were 66.53 +/- 32.22 and 51.10 +/- 18.29 milliseconds. With these 3 methods, the postoperative findings were significantly lower than the preoperative findings ( P < .05 for all). In contrast, the opposite was true with the QT CB method (preoperative 71.1 +/- 65.80 vs postoperative 76.43 +/- 7.96 milliseconds, P < .05). CONCLUSION: The study showed that based on the methods of Fridericia (QT CF ) and Sagie et al (QT CS ), QT intervals are significantly decreased after CABG and aneurysmectomy. PMID- 15894978 TI - Why do patients fail to receive beta-blockers for chronic heart failure over time? A "real-world" single-center, 2-year follow-up experience of beta-blocker therapy in patients with chronic heart failure. AB - BACKGROUND: The longitudinal pattern of beta-blocker use in a heart-failure practice setting has not been explored. Previous studies have not specifically addressed the use of beta-blockers over time to determine the rate of use and reasons for discontinuation. The long-term compliance rate for beta-blocker drugs outside the context of a clinical trial has not been established. METHODS: We prospectively followed a cohort of 500 consecutive patients between March and May 2001, with a clinical diagnosis of chronic heart failure seen in a specialized heart failure clinic and determined the longitudinal pattern of beta-blocker use and clinical outcomes over a 2-year period. RESULTS: The final cohort consists of 340 patients with a complete 2-year follow-up data (mean age 61 +/- 14 years, 69% men, 53% with ischemic etiology, mean ejection fraction 27.6 +/- 15%). At 6, 12, and 24 months, beta-blocker utilization rates were maintained in 69%, 70%, and 74% of patients, respectively. Of the 120 confirmed initial non-beta-blocker users, 28 (23%) were subsequently started on beta-blocker, despite suspected relative contraindications in 53% of patients. Over a period of 2 years, the discontinuation rate was 10%, with failure to restart a beta-blocker after hospitalization as the most common reason for beta-blocker discontinuation. CONCLUSION: Utilization rates of beta-blockers in our heart failure clinic have remained constant at approximately 70% throughout a 2-year follow-up. Of those who discontinued beta-blockers (10%), the most common documented cause was failure to restart beta-blockers after hospitalization. PMID- 15894979 TI - Hypothermia is a bedside predictor of imminent death in patients with congestive heart failure. AB - BACKGROUND: Most studies on predictors of mortality for patients with congestive heart failure (CHF) have described predictors that are either difficult to measure in routine practice or are only modestly sensitive and specific. Having observed 3 patients whose body temperature decreased shortly before death, we hypothesized that hypothermia may predict inhospital mortality. METHODS: The medical records of 291 patients with a primary discharge diagnosis of CHF were selected from 423 admissions to Memorial Hermann Hospital, Houston, Tex, 1998, after excluding patients with comorbidities that confound body temperature, deaths for causes other than progressive pump failure, and readmissions except the last. Three groups were defined on the basis of admission body temperature (T adm): hypothermia groups T adm (95.5 degrees F-96.5 degrees F) and T adm < 95.5 degrees F, and reference group T adm > or = 96.6 degrees F. Several other known CHF risk factors were studied for confounding, and adjusted hazard ratios were calculated using Cox regression. RESULTS: Of the 291 patients (mean age 73 years, 47% men), 17 (6%) had hypothermia on admission. Mean hospital stay was 5 days. Of the 17 (6%) patients who died of pump failure, 5 had been hypothermic on admission. Hypothermia was significantly associated with survival, and after adjusting for New York Heart Association functional class, hazard ratio for T adm < 95.5 degrees F was 4.46 (95% confidence interval 1.38-14.3) (P trend = .0283). CONCLUSIONS: Hypothermia predicted inhospital death in these patients with CHF. If confirmed by future studies, this finding could prove useful, because temperature can be measured continuously, rapidly, and inexpensively, in or out of the hospital. PMID- 15894980 TI - Blood pressure variability and prognosis in uncomplicated mild hypertension. AB - BACKGROUND: The independent prognostic value of blood pressure (BP) variability in essential hypertension and particularly in uncomplicated mild hypertension is not yet completely clear. We investigated the relationship between BP variability, evaluated by noninvasive monitoring, and cardiovascular outcome in uncomplicated mild hypertension patients. METHODS: The occurrence of fatal and nonfatal cardiovascular events was evaluated in 1088 uncomplicated mild hypertension patients, 626 with low BP variability (SD of daytime systolic BP below the mean of the study population) and 462 with high BP variability (SD of daytime systolic BP above the mean of the study population). RESULTS: During the follow-up (4.74 +/- 2.4 years, range 0.5-9.7 years), the event rates per 100 patient-years in subjects with low and high BP variability were 0.72 and 1.5, respectively. Event-free survival was significantly different between the groups (P = .01). However, after adjustment for other covariates in a Cox multivariate analysis, the adverse prognostic relevance of high BP variability was no longer detectable, whereas age, daytime systolic BP, low-density lipoprotein cholesterol, and smoking habit resulted as independent predictors of cardiovascular events. CONCLUSIONS: In uncomplicated mild hypertension population, BP variability evaluated by noninvasive monitoring is not an independent predictor of outcome. PMID- 15894981 TI - The data monitoring experience in the Candesartan in Heart Failure Assessment of Reduction in Mortality and morbidity (CHARM) program. AB - BACKGROUND: The CHARM program was designed as 3 separate randomized trials comparing candesartan with placebo in patients with chronic heart failure (CHF) who (1) were intolerant to angiotensin-converting enzyme inhibitor and had left ventricular ejection fraction (LVEF) < or =0.40; (2) were on angiotensin converting enzyme inhibitor and had LVEF < or =0.40; or (3) had LVEF >0.40. CHARM provides an interesting example of the challenges faced by a Data and Safety Monitoring Committee (DSMC). METHODS: Although the primary efficacy end point for each component trial was cardiovascular (CV) death or hospitalization for CHF, the primary outcome for the overall program was all-cause mortality. The DSMC received monthly safety reports and also met every 6 months (7 times in all) to review interim reports. Statistical stopping guidelines were predefined for all cause mortality in the overall program. The overarching principle of the DSMC was proof beyond a reasonable doubt that would be likely to influence clinical practice. RESULTS: There were significant treatment differences in all-cause mortality for the overall program at several interim analyses, and the statistical stopping guideline was reached on one occasion. However, even a conventional level of statistical significance (P < .050) was achieved in only 1 of the 3 component trials. The DSMC consistently recommended that the program continue as planned. The final published result for all-cause death over a median of 3.1 years was a 9% reduction in hazard (95% CI 0%-17%, P = .055), whereas for CV death or hospitalization for CHF, there was a 16% reduction in hazard (95% CI 9%-23%, P < .0001). Subsequent exploratory analyses suggest that the hazard reduction in CV death was more marked in the first year after randomization and that, if real, this apparent treatment-time interaction offers a plausible explanation for why the interim mortality data showed statistically more extreme findings than the overall final results. CONCLUSION: The DSMC experience in the CHARM program illustrates the importance of continuing a trial to its scheduled completion unless there is proof beyond a reasonable doubt that would influence clinical practice rather than strict reliance on a statistical stopping guideline. PMID- 15894993 TI - The tip of the genomic health care iceberg. PMID- 15894994 TI - Genetic competencies essential for health care professionals in primary care. AB - The completion of the sequencing of the human genome in 2003 signaled the onset of the genomic era in health care. The knowledge gleaned from the Human Genome Project has led to the understanding that every health problem has a genetic component and that clinicians should include the application of genetic information in all aspects of health care. This article describes the genetic competencies essential for all health care professionals in primary care. Health care professionals should augment their current practice by obtaining a multigenerational genetic family history for each patient, assessing all patients for potentially heritable conditions, providing referrals to genetic health professionals as needed, offering genetic testing when indicated, and considering an individual's genetic makeup in the selection of medications and treatments for that person. Finally, all health care professionals ought to be prepared to address the complex personal, cultural, theological, ethical, legal, and social issues associated with genetic testing and other genetic issues commonly encountered in clinical practice. PMID- 15894996 TI - Harnessing the power of the pedigree. AB - Primary care providers are in an ideal position to practice genomic-based medicine. Family history data can be used to assess reproductive risks or determine an individual's risk for developing specific diseases. The US Department of Health and Human Services has recently launched the US Surgeon General's Family History Initiative, a national public health campaign designed to encourage Americans to learn more about their family health histories. Furthermore, several national associations now recommend that primary care providers collect family history data to identify patients at risk for these diseases. Ideally, family history data should be ascertained, documented, and analyzed in a standardized manner. Graphic representation of a family history in the form of a pedigree may be preferable to a text format, but further research will clarify this issue. Family history tools are now being developed and studied to identify which methods are most beneficial in different clinical settings. PMID- 15894995 TI - Speaking the language of genetics: a primer. AB - Genetic research advances will continue to result in clinical applications for genetics in primary care settings. Fluency with the evolving genetic terminology will enable primary care providers to provide better clinical care to their patients, particularly when helping patients understand genetic concepts. This article will help clinicians use genetic terminology with greater precision. PMID- 15894997 TI - A primer on genetic testing. AB - Use of genetic tests in the clinical practice setting is a current reality, and an increasing number of patients are asking about and requesting genetic testing. The push to translate genetic research findings and technological innovations into clinical practice will continue as our understanding of the genetic basis of disease increases. Special consideration is required when ordering genetic tests, beyond that of other laboratory tests, and an understanding of the unique aspects involved will help optimize clinical outcomes. The purpose of this primer is to provide a basic understanding of genetic testing, discuss current issues related to the use of tests, and outline practical steps for critically using genetic tests in clinical practice. PMID- 15894998 TI - Cystic fibrosis screening. AB - The integration of universal cystic fibrosis screening in women's health has considerably altered the way we care for obstetric patients and likely will be the foundation for incorporating other genetic tests into routine women's health care. Prior to this change in the standard of care, screening for genetic disease was primarily limited to those individuals who had a personal or family history of the genetic condition or who belonged to a particular ethnic or racial group characterized by a high frequency of carrier and affected individuals. However, technological advances have resulted in facile and economic methodologies for detecting an increasing number of genetic mutations and in the realization that screening for common and uncommon disorders will likely be a not-too-distant future part of routine health care. Programs that permit clinicians to properly implement genetic protocols and allow patients to make informed decisions about genetic screening and diagnostic tests are needed. The implementation of universal cystic fibrosis screening allows clinicians to recognize the benefits and pitfalls of genetic testing of the general population and encourages the development of programs that will effectively communicate genetic information to professionals and laity. PMID- 15894999 TI - Options for Down syndrome screening: what will women choose? AB - Down syndrome screening has been offered to pregnant women since the early 1980s. Protocols have changed as research confirmed improvements that result in higher detection rates and lower false-positive rates. Results from 2 clinical trials evaluating screening protocols that include ultrasound measurement of nuchal translucency and biochemical testing in the first and second trimester are now available. First-trimester screening is an option if there are adequate ultrasound, diagnostic, and counseling services available. Regional variation in the availability of these services may limit the implementation of first trimester screening. Combining screening tests for Down syndrome from both trimesters as an integrated test offers the highest detection rate with the lowest false-positive rate. The possibility of avoiding a positive screen will make this an attractive option for some. Timing, detection rate, false-positive rates, and personal factors influence the decision women make regarding screening versus diagnostic testing. This article reviews the efficacy of current protocols for Down syndrome screening. Accurate information about available screening tests will facilitate informed decisions about screening and testing. PMID- 15895000 TI - Newborn screening and genetic testing. AB - New screening techniques and diagnostic tests for genetic diseases available for newborn screening can provide information about many diseases long before they are clinically detected. However, this information creates complex questions and ethical dilemmas regarding which newborns should be tested, when testing should occur, availability and costs of tests, and how families should be counseled. There is no national policy regarding newborn screening, which leads to great variation among states' newborn screening programs. This article reviews newborn genetic testing and provides a blueprint for clinicians to improve practice by incorporating into their care knowledge of new developments in newborn testing and screening. PMID- 15895001 TI - Genetic testing and the family. AB - The family experience of genetic testing is explored in this article. Two family stories are presented to illustrate how families define and manage the ethical and social issues that emerge during 2 types of genetic testing: mutation analysis for Huntington's disease and genetic testing for breast and ovarian cancer susceptibility. These 2 families were purposefully selected because their stories exemplify the complexity of the genetic testing experience. In addition, the story of the family living with Huntington's disease shows how negative consequences can occur for the individual tested, other family members, the marital relationship, and the family system, even when the test results indicate that the individual does not carry a deleterious gene mutation. Both of the families presented in this article participated in an ongoing study, Family Experience of Genetic Testing: Ethical Dimensions , in which 118 family members from 67 families have participated. The guiding framework for this research was the family management style framework developed and refined by Knafl and colleagues. PMID- 15895002 TI - Ethical issues in genetic testing. AB - Genetic factors contribute to risk for disease. Information clarifying risk in an individual and his/her family members can be identified through clinical screening and genetic testing. In some circumstances, this information can be used in clinical decisions about surveillance and prevention or treatment of disease. However, use of this information is not always a straightforward process. Application of knowledge about risk of genetic disease in provision of primary health care for women requires understanding of new genetic discoveries as well as the ability to participate in resolution of ethical dilemmas that may result when genetic screening and testing are considered. These dilemmas arise not only from the current state of knowledge about genetic risk factors and utility of genetic tests but also result from conflicts that can arise when the needs of the client are not in unison with needs of others within the family or society. Ethical theories and principles provide a framework for resolving ethical dilemmas in maternal screening for genetic conditions during a pregnancy, carrier testing prior to or during a pregnancy, clinical genetic testing, and newborn metabolic screening. PMID- 15895003 TI - Special children. PMID- 15895004 TI - Incidental finding of ultrasound markers for Down syndrome in the second trimester of pregnancy: a case study. PMID- 15895005 TI - Genetic resources for midwifery practice. PMID- 15895013 TI - Prenatal tests for Down syndrome. PMID- 15895014 TI - The integration of feminism and midwifery. PMID- 15895016 TI - Mary Breckenridge and the birth of ACNM. PMID- 15895020 TI - Tribute to Teresa Marsico, CNM, MEd, FACNM (1937-2005). PMID- 15895022 TI - Treatment of bitumen burns: effective dissolution of hardened hydrocarbon residue on periorbital and eyelid burns using butter. PMID- 15895023 TI - Prevalence of pterygium and cataract in indigenous populations of the Brazilian Amazon rain forest. AB - PURPOSE: To compare the prevalence of pterygium and cataract in four indigenous populations of the Brazilian Amazonian rain forest (Arawak, Tukano, Maku, and Yanomami) with different ethnic and social behaviour backgrounds. METHODS: A cross-sectional pterygium and cataract survey was performed in 624 adult Indians of the Brazilian rain forest belonging to four different ethnic groups. The Indians were classified according to their social behaviour in two groups: Arawak and Tukano (group 1) and Maku and Yanomami (group 2). Slit-lamp biomicroscopy was employed to examine the entire sample. All subjects were classified as 1 or 0 according to the presence or absence pterygium and cataract. Sex and age were also recorded. RESULTS: chi(2)-tests revealed that the prevalence of pterygium and cataract differed significantly between groups 1 and 2. For pterygia: 36.6% (97/265) and 5.0% (18/359), respectively (chi(2)=101.2, P<0.0001), and for cataracts: 24.5% (65/265) and 13.7% (49/359) respectively (chi(2)=12.09, P=0.0005). Gender was not associated with pterygium (P=0.1326) and cataract (P=0.2263) in both groups. Elderly subjects showed a significantly higher prevalence of cataract (P<0.0001). The prevalence of pterygia did not increase with age (P=0.8079) in both groups. CONCLUSION: Indians of group 1 have higher prevalence of pterygia and cataract than Indians of group 2. Social behaviour, especially the rate of sun exposure, appears to be the main factor for the different rates of pterygium and cataract displayed by these indigenous people of the Brazilian rain forest. PMID- 15895024 TI - Screening for age-related macular degeneration using nonstereo digital fundus photographs. AB - Age-related macular degeneration (AMD) is a disease with significant visual morbidity and accounts for the majority of blind registrations in the developed world including the UK. Certain forms of neovascular AMD are amenable to treatment but require expeditious referral to a retinal specialist. AIM: To evaluate the possibility of using nonstereo fundus photographs as a low-cost screening tool for neovascular AMD. DESIGN: Retrospective review of patients referred to the macular clinic of a teaching hospital in London. METHODS: A total of 198 randomised digital fundus photographs, without any other clinical information, were presented to two independent ophthalmic interns who graded them into one of the three categories: normal, age-related maculopathy (ARM), or neovascular age-related macular degeneration (AMD) to determine the urgency of referral to clinic. The results were compared with the known diagnosis for each patient and sensitivities and specificities for each diagnostic category calculated. RESULTS: The intraobserver Kappa statistic was 0.75 and 0.91 for grader 1 and 2, respectively. The interobserver Kappa was 0.54. The mean sensitivity and specificity for the identification of ARM was 60.5 and 76.3%, respectively The mean sensitivity and specificity for the identification of AMD was 85.7 and 78.8%, respectively. CONCLUSION: Nonstereo digital fundus photograph is a reasonable screening tool for CNV and may aid in decreasing the visual morbidity it causes by enabling timely referrals and treatment. PMID- 15895025 TI - The usefulness of the Amsler chart. PMID- 15895026 TI - Red dots visual field test with blue on yellow & blue on red macula test grid. PMID- 15895028 TI - Neuroblastoma with orbital metastasis: ophthalmic presentation and role of ophthalmologists. AB - INTRODUCTION: Neuroblastoma is predominantly a tumour of early childhood, which metastasises to the orbits. In such cases, ophthalmologists are involved in the multidisciplinary management. This unique series from a tertiary referral centre is used to elaborate the ophthalmic associations and the ophthalmologist's role in this rare condition. METHODS: A review of case notes was performed on six patients who presented to the paediatric ophthalmology - oncology liaison service at the Leeds teaching hospitals between 1998 and 2003. The ophthalmic outcome and role of the ophthalmologist were assessed. RESULTS: Average age of presentation was 29.8 months (range 15-69 months). Average duration of follow-up was 19.5 months (range 2-58 months). One child died during treatment. Two have completed treatment and are under follow-up. Presenting features of the six children were proptosis in four, periorbital ecchymosis in two, ocular motility restriction in two, and subconjunctival haemorrhage in one. Only one case developed blindness. CONCLUSIONS: The role of the ophthalmologist in patients with metastatic orbital neuroblastoma can vary from a supportive role to one of active intervention and management of ophthalmic complications. The ophthalmologist is involved in diagnosis and staging as well as monitoring response to treatment of both the primary disease and secondary ophthalmic complications. PMID- 15895027 TI - Mesenchymal stem cell transplantation in a rabbit corneal alkali burn model: engraftment and involvement in wound healing. AB - PURPOSE: To investigate whether systemically transplanted mesenchymal stem cells (MSCs) can home and engraft in tissue to promote cornea wound healing after alkali burn, as a new source for treatment. METHODS: Corneal alkali burn was created in four group rabbits: Group I, normal bone marrow function, without MSCs transplantation; Group II, normal bone marrow function, with MSCs transplantation; Group III, bone marrow suppressed by cyclophosphamide, without MSCs; Group IV, bone marrow suppressed by cyclophosphamide, with MSCs. Clinical outcome was evaluated by cornea re-epithelization, cornea opacity, and neovascularization. Cell engraftment into bone marrow, circulation, and cornea was monitored. Immunohistochemistry, using proliferating cell nuclear antigen (PCNA), P63, vimentin, and alpha-smooth muscle actin (alpha-SMA) was carried out to assess the cell proliferative and differentiative ability. RESULTS: At the time of 1-month follow-up, Group II rabbits showed the best clinical results with a clearer healed cornea compared with other groups. Well-formed neovascularization appeared on day 14 after alkali burn in Group II, that coincided with the maximum engraftment of MSCs. PCNA, P63, vimentin were more strongly expressed in Group II at multiple time points. DiI-labelled MSCs were differentiated into myofibroblast by the expression of alpha-SMA. Delayed and insufficient cell engraftment, with malformed neovascularization and retarded corneal wound healing was found in Groups III and IV. CONCLUSIONS: Systemically transplanted MSCs can engraft to injured cornea to promote wound healing, by differentiation, proliferation, and synergizing with haemotopoietic stem cells. PMID- 15895029 TI - Optical coherence tomography of fundus findings in type II mesangiocapillary glomerulonephritis. PMID- 15895030 TI - Bilateral jugular vein thrombosis: a rare cause of papilloedema. PMID- 15895031 TI - Comment on 'quantification of the role of temporal artery biopsy in diagnosing clinically suspected giant cell arteritis'. PMID- 15895032 TI - What's the point? PMID- 15895033 TI - Dental project. PMID- 15895034 TI - Injury assessing. PMID- 15895035 TI - Foreign dentists. PMID- 15895044 TI - The dental role in smoking cessation advice for patients with mental and behavioural disorders--time for improvement? AB - The past few years have seen an increasing emphasis upon the role of the dental team in smoking cessation. The recent advances in knowledge of the level and role of tobacco use in patients with mental and behavioural disorders, the biological factors involved and the influences of tobacco on psychiatric medication metabolism and side effects has not been reflected in dental postgraduate education. Perhaps it is time for an evidence based improvement in care standards by referring these patients for specialist cessation advice? PMID- 15895045 TI - Post and core systems, refinements to tooth preparation and cementation. AB - With a plethora of post systems available, it is often difficult to decide which one to use. This is made more difficult by the fact that new posts are introduced before existing ones are fully evaluated in laboratory and clinical studies. This paper therefore describes the different post types and the main advantages and disadvantages of each. In addition, the choice of post system will influence whether further tooth preparation is required and will dictate which luting cement and core material are most appropriate. Whilst the choice of post will, for many dentists, be driven by personal preference and a history of clinical success, there are certain pit falls to avoid and these are outlined. PMID- 15895046 TI - An extraction complicated by lateral and medial pterygoid tethering of a fractured maxillary tuberosity. AB - We report a case in which the extraction of an upper second molar was complicated by a maxillary tuberosity fracture. Delivery of the tooth and bone fragment under local anaesthesia was unable to be achieved because of pain, brisk bleeding and tethering by the lateral and medial pterygoid muscles. The eventual removal of the fragment under general anaesthetic required the control of haemorrhage deep within the infratemporal fossa. When this complication is recognised by the general dentist the maxillary tuberosity should not be removed and the patient referred to a specialist unit. PMID- 15895047 TI - Mediastinitis from odontogenic infection. A case report. AB - We report a case of mediastinitis complicating a dental infection in a 40-year old male. Despite drainage of the localised neck abscess and the administration of systemic antibiotics, his submandibular abscess extended to involve the pericardial and pleural cavities. Drainage procedures and thoracotomies were required to treat the empyema and purulent pericarditis. Computed tomography was used to follow the progression of disease and assess the efficacy of treatment. PMID- 15895048 TI - The Palmer notation system and its use with personal computer applications. AB - The arguments for and against the Palmer dental notation system are briefly discussed, including the perceived difficulty of reproducing this on personal computers. Some technical solutions to the problems encountered in everyday Windows-based programs are outlined, with suggestions as to possible future applications. PMID- 15895055 TI - Classifications. PMID- 15895056 TI - Potential toxicity. PMID- 15895057 TI - A comparison of two radiographic assessment protocols for patients with periodontal disease. AB - OBJECTIVE: Radiographic assessment of patients with generalised severe periodontitis may be undertaken with a panoramic view and supplementary periapicals. The purpose of this study was to estimate the effective radiation dose from this form of radiographic assessment, and to compare it with an estimate of the dose from a series of periapicals of all the affected teeth. DESIGN: Cross-sectional observational study. SETTING: Departments of Periodontology and Radiology, Glasgow Dental Hospital and School. METHOD: Fifty consecutive patients [were recruited] with sufficiently widespread advanced periodontitis to require at least seven periapical radiographs. [Following new local guidelines, a panoramic view was taken.] The adequacy of the image of every affected tooth and the number of supplementary periapicals required was determined by a panel of four examiners who also calculated the number of periapicals which would have been taken if panoramic radiography had not been available. An effective dose of 0.001 mSv for one periapical and 0.007 mSv for a panoramic view was assumed. RESULTS: The panoramic-plus-periapicals approach delivered an estimated additional effective dose to 86% of patients, in the order of 0.001-0.007 mSv. CONCLUSIONS: Within the parameters of this investigation, the anticipated effective radiation dose from a series of periapical radiographs of all selected teeth would, for the great majority of patients, have been less than the dose from a panoramic-plus-periapicals approach. PMID- 15895059 TI - Fairy tales. AB - Everyone knows the story of the tooth fairy, but fine art degree student Cordelia Cembrowicz has transformed the mythical figure in a most unusual way. PMID- 15895058 TI - Views and experiences of parents and siblings of adults with Down Syndrome regarding oral healthcare: a qualitative and quantitative study. AB - AIMS: To investigate experiences and expectations of parents/siblings of adults with Down Syndrome (DS) regarding oral healthcare, and explore factors impacting on access and experience of dental care for this group. DESIGN: A two phase qualitative and quantitative study using in-depth interviews with a convenience sample of six parents/siblings, and a postal questionnaire of 200 parents/siblings of adults with DS who are members of the Down Syndrome Association. RESULTS: The main themes elicited from the qualitative interviews related to concern, experiences, parents'/siblings' attitudes, preferences and information. The response rate from the postal questionnaire was 63.5%. Adults with DS attended the dentist regularly but received little restorative treatment. Experience of oral healthcare was influenced by the attitudes and skills of dental health professionals; stigma; and relatives' expectations of dentists, their oral health beliefs, information and support received, knowledge and priorities. Parents/siblings wanted dentists to be proactive in providing more information on oral health issues in collaboration with other health and social care professionals. CONCLUSIONS: Whilst most adults with DS visited the dentist regularly, relatively little treatment had been provided. Parents highlighted a need for appropriate and timely oral health information early in their child's life, and access to dentists who were sympathetic, good communicators and well informed about DS. PMID- 15895072 TI - Mycobacterium tuberculosis isocitrate lyases 1 and 2 are jointly required for in vivo growth and virulence. AB - Genes involved in fatty acid catabolism have undergone extensive duplication in the genus Mycobacterium, which includes the etiologic agents of leprosy and tuberculosis. Here, we show that prokaryotic- and eukaryotic-like isoforms of the glyoxylate cycle enzyme isocitrate lyase (ICL) are jointly required for fatty acid catabolism and virulence in Mycobacterium tuberculosis. Although deletion of icl1 or icl2, the genes that encode ICL1 and ICL2, respectively, had little effect on bacterial growth in macrophages and mice, deletion of both genes resulted in complete impairment of intracellular replication and rapid elimination from the lungs. The feasibility of targeting ICL1 and ICL2 for chemical inhibition was shown using a dual-specific ICL inhibitor, which blocked growth of M. tuberculosis on fatty acids and in macrophages. The absence of ICL orthologs in mammals should facilitate the development of glyoxylate cycle inhibitors as new drugs for the treatment of tuberculosis. PMID- 15895073 TI - Stat3 is required for ALK-mediated lymphomagenesis and provides a possible therapeutic target. AB - Anaplastic large cell lymphomas (ALCLs) are caused by chromosomal translocations that juxtapose the anaplastic lymphoma kinase (ALK) proto-oncogene to a dimerization partner, resulting in constitutive expression of ALK and ALK tyrosine kinase activity. One substrate of activated ALK in human ALCLs is the transcription factor Stat3, and its phosphorylation is accurately recapitulated in a new nucleophosmin (NPM)-ALK transgenic mouse model of lymphomagenesis. Here we show by gene targeting that Stat3 is required for the transformation of mouse embryonic fibroblasts in vitro, for the development of B-cell lymphoma in transgenic mice and for the growth and survival of both human and mouse NPM-ALK transformed B and T cells. Ablation of Stat3 expression by antisense oligonucleotides significantly (P < 0.0001) impaired the growth of human and mouse NPM-ALK tumors in vivo. Pharmacological ablation of Stat3 represents a new candidate approach for the treatment of human lymphoma PMID- 15895074 TI - Inflammatory arthritis requires Foxo3a to prevent Fas ligand-induced neutrophil apoptosis. AB - In inflammatory arthridities such as rheumatoid arthritis, cognate lymphocytes have long been considered instigators of autoimmunity, but accumulating evidence indicates that innate immune cells such as neutrophils and mast cells are responsible for a vast majority of acute and ongoing inflammation; however, the molecular mechanisms that govern them remain largely unknown. Here we show that such inflammation requires the forkhead transcription factor Foxo3a: Foxo3a deficient mice are resistant to two models of neutrophilic inflammation, immune complex-mediated inflammatory arthritis and thioglycollate-induced peritonitis. This reflects a need for Foxo3a to maintain neutrophil vitality during inflammation by suppressing Fas ligand; because Foxo3a can bind and suppress the Fasl promoter, Foxo3a-deficient neutrophils upregulate Fas ligand and undergo apoptosis in response to TNF-alpha and IL-1, and Fas ligand blockade renders Foxo3a-deficient mice susceptible to both arthritis and peritonitis. Thus, Foxo3a ensures neutrophil survival during inflammation, identifying Foxo3a as therapeutic target in inflammation. PMID- 15895075 TI - Intermolecular complementation achieves high-specificity tumor targeting by anthrax toxin. AB - Anthrax toxin protective antigen (PrAg) forms a heptamer in which the binding site for lethal factor (LF) spans two adjacent monomers. This suggested that high cell-type specificity in tumor targeting could be obtained using monomers that generate functional LF-binding sites only through intermolecular complementation. We created PrAg mutants with mutations affecting different LF-binding subsites and containing either urokinase plasminogen activator (uPA) or matrix metalloproteinase (MMP) cleavage sites. Individually, these PrAg mutants had low toxicity as a result of impaired LF binding, but when administered together to uPA- and MMP-expressing tumor cells, they assembled into functional LF-binding heteroheptamers. The mixture of two complementing PrAg variants had greatly reduced toxicity in mice and was highly effective in the treatment of aggressive transplanted tumors of diverse origin. These results show that anthrax toxin, and by implication other multimeric toxins, offer excellent opportunities to introduce multiple-specificity determinants and thereby achieve high therapeutic indices. PMID- 15895076 TI - Microtubule-induced focal adhesion disassembly is mediated by dynamin and focal adhesion kinase. AB - Imaging studies implicate microtubule targeting of focal adhesions in focal adhesion disassembly, although the molecular mechanism is unknown. Here, we develop a model system of focal adhesion disassembly based on the finding that microtubule regrowth after nocodazole washout induces disassembly of focal adhesions, and that this disassembly occurs independently of Rho and Rac, but depends on focal adhesion kinase (FAK) and dynamin. During disassembly, dynamin interacts with FAK and colocalizes with focal adhesions. Inhibition of dynamin prevents migration of cells with a focal adhesion phenotype. Our results show that focal adhesion disassembly involves microtubules, dynamin and FAK, and is not simply the reversal of focal adhesion formation. PMID- 15895077 TI - Caenorhabditis elegans RME-6 is a novel regulator of RAB-5 at the clathrin-coated pit. AB - Here we identify a new regulator of endocytosis called RME-6. RME-6 is evolutionarily conserved among metazoans and contains Ras-GAP (GTPase-activating protein)-like and Vps9 domains. Consistent with the known catalytic function of Vps9 domains in Rab5 GDP/GTP exchange, we found that RME-6 binds specifically to Caenorhabditis elegans RAB-5 in the GDP-bound conformation, and rme-6 mutants have phenotypes that indicate low RAB-5 activity. However, unlike other Rab5 associated proteins, a rescuing green fluorescent protein (GFP)-RME-6 fusion protein primarily localizes to clathrin-coated pits, physically interacts with alpha-adaptin, a clathrin adaptor protein, and requires clathrin to achieve its cortical localization. In rme-6 mutants, transport from the plasma membrane to endosomes is defective, and small 110-nm endocytic vesicles accumulate just below the plasma membrane. These results suggest a mechanism for the activation of Rab5 in clathrin-coated pits or clathrin-coated vesicles that is essential for the delivery of endocytic cargo to early endosomes. PMID- 15895078 TI - Prediction of preadipocyte differentiation by gene expression reveals role of insulin receptor substrates and necdin. AB - The insulin/IGF-1 (insulin-like growth factor 1) signalling pathway promotes adipocyte differentiation via complex signalling networks. Here, using microarray analysis of brown preadipocytes that are derived from wild-type and insulin receptor substrate (Irs) knockout animals that exhibit progressively impaired differentiation, we define 374 genes/expressed-sequence tags whose expression in preadipocytes correlates with the ultimate ability of the cells to differentiate. Many of these genes, including preadipocyte factor-1 (Pref-1) and multiple members of the Wnt signalling pathway, are related to early adipogenic events. Necdin is also markedly increased in Irs knockout cells that cannot differentiate, and knockdown of necdin restores brown adipogenesis with downregulation of Pref-1 and Wnt10a expression. Insulin receptor substrate proteins regulate a necdin-E2F4 interaction that represses peroxisome proliferator-activated receptor gamma (PPARgamma) transcription via a cyclic AMP response element binding protein (CREB)-dependent pathway. Together these define a key signalling network that is involved in brown preadipocyte determination. PMID- 15895079 TI - Alternative splicing and gene duplication are inversely correlated evolutionary mechanisms. AB - Gene duplication and alternative splicing are distinct evolutionary mechanisms that provide the raw material for new biological functions. We explored their relationships in human and mouse and found an inverse correlation between the size of a gene's family and its use of alternatively spliced isoforms. A cross organism analysis suggests that selection for genome-wide genic proliferation might be interchangeably met by either evolutionary mechanism. PMID- 15895080 TI - Developmental stage-selective effect of somatically mutated leukemogenic transcription factor GATA1. AB - Acquired mutations in the hematopoietic transcription factor GATA binding protein 1 (GATA1) are found in megakaryoblasts from nearly all individuals with Down syndrome with transient myeloproliferative disorder (TMD, also called transient leukemia) and the related acute megakaryoblastic leukemia (DS-AMKL, also called DS-AML M7). These mutations lead to production of a variant GATA1 protein (GATA1s) that is truncated at its N terminus. To understand the biological properties of GATA1s and its relation to DS-AMKL and TMD, we used gene targeting to generate Gata1 alleles that express GATA1s in mice. We show that the dominant action of GATA1s leads to hyperproliferation of a unique, previously unrecognized yolk sac and fetal liver progenitor, which we propose accounts for the transient nature of TMD and the restriction of DS-AMKL to infants. Our observations raise the possibility that the target cells in other leukemias of infancy and early childhood are distinct from those in adult leukemias and underscore the interplay between specific oncoproteins and potential target cells. PMID- 15895081 TI - Epistasis between mouse Klra and major histocompatibility complex class I loci is associated with a new mechanism of natural killer cell-mediated innate resistance to cytomegalovirus infection. AB - Experimental infection with mouse cytomegalovirus (MCMV) has been used to elucidate the intricate host-pathogen mechanisms that determine innate resistance to infection. Linkage analyses in F(2) progeny from MCMV-resistant MA/My (H2 (k)) and MCMV-susceptible BALB/c (H2 (d)) and BALB.K (H2 (k)) mouse strains indicated that only the combination of alleles encoded by a gene in the Klra (also called Ly49) cluster on chromosome 6, and one in the major histocompatibility complex (H2) on chromosome 17, is associated with virus resistance. We found that natural killer cell-activating receptor Ly49P specifically recognized MCMV-infected cells, dependent on the presence of the H2 (k) haplotype. This binding was blocked using antibodies to H-2D(k) but not antibodies to H-2K(k). These results are suggestive of a new natural killer cell mechanism implicated in MCMV resistance, which depends on the functional interaction of the Ly49P receptor and the major histocompatibility complex class I molecule H-2D(k) on MCMV-infected cells. PMID- 15895082 TI - Epigenome analyses using BAC microarrays identify evolutionary conservation of tissue-specific methylation of SHANK3. AB - CpG islands are present in one-half of all human and mouse genes and typically overlap with promoters or exons. We developed a method for high-resolution analysis of the methylation status of CpG islands genome-wide, using arrays of BAC clones and the methylation-sensitive restriction enzyme NotI. Here we demonstrate the accuracy and specificity of the method. By computationally mapping all NotI sites, methylation events can be defined with single-nucleotide precision throughout the genome. We also demonstrate the unique expandability of the array method using a different methylation-sensitive restriction enzyme, BssHII. We identified and validated new CpG island loci that are methylated in a tissue-specific manner in normal human tissues. The methylation status of the CpG islands is associated with gene expression for several genes, including SHANK3, which encodes a structural protein in neuronal postsynaptic densities. Defects in SHANK3 seem to underlie human 22q13 deletion syndrome. Furthermore, these patterns for SHANK3 are conserved in mice and rats. PMID- 15895083 TI - Fine-scale structural variation of the human genome. AB - Inversions, deletions and insertions are important mediators of disease and disease susceptibility. We systematically compared the human genome reference sequence with a second genome (represented by fosmid paired-end sequences) to detect intermediate-sized structural variants >8 kb in length. We identified 297 sites of structural variation: 139 insertions, 102 deletions and 56 inversion breakpoints. Using combined literature, sequence and experimental analyses, we validated 112 of the structural variants, including several that are of biomedical relevance. These data provide a fine-scale structural variation map of the human genome and the requisite sequence precision for subsequent genetic studies of human disease. PMID- 15895084 TI - ATP mediates rapid microglial response to local brain injury in vivo. AB - Parenchymal microglia are the principal immune cells of the brain. Time-lapse two photon imaging of GFP-labeled microglia demonstrates that the fine termini of microglial processes are highly dynamic in the intact mouse cortex. Upon traumatic brain injury, microglial processes rapidly and autonomously converge on the site of injury without cell body movement, establishing a potential barrier between the healthy and injured tissue. This rapid chemotactic response can be mimicked by local injection of ATP and can be inhibited by the ATP-hydrolyzing enzyme apyrase or by blockers of G protein-coupled purinergic receptors and connexin channels, which are highly expressed in astrocytes. The baseline motility of microglial processes is also reduced significantly in the presence of apyrase and connexin channel inhibitors. Thus, extracellular ATP regulates microglial branch dynamics in the intact brain, and its release from the damaged tissue and surrounding astrocytes mediates a rapid microglial response towards injury. PMID- 15895085 TI - Ovarian cycle-linked changes in GABA(A) receptors mediating tonic inhibition alter seizure susceptibility and anxiety. AB - Disturbances of neuronal excitability changes during the ovarian cycle may elevate seizure frequency in women with catamenial epilepsy and enhance anxiety in premenstrual dysphoric disorder (PMDD). The mechanisms underlying these changes are unknown, but they could result from the effects of fluctuations in progesterone-derived neurosteroids on the brain. Neurosteroids and some anxiolytics share an important site of action: tonic inhibition mediated by delta subunit-containing GABA(A) receptors (deltaGABA(A)Rs). Here we demonstrate periodic alterations in specific GABA(A)R subunits during the estrous cycle in mice, causing cyclic changes of tonic inhibition in hippocampal neurons. In late diestrus (high-progesterone phase), enhanced expression of deltaGABA(A)Rs increases tonic inhibition, and a reduced neuronal excitability is reflected by diminished seizure susceptibility and anxiety. Eliminating cycling of deltaGABA(A)Rs by antisense RNA treatment or gene knockout prevents the lowering of excitability during diestrus. Our findings are consistent with possible deficiencies in regulatory mechanisms controlling normal cycling of deltaGABA(A)Rs in individuals with catamenial epilepsy or PMDD. PMID- 15895086 TI - Subunit interaction with PICK and GRIP controls Ca2+ permeability of AMPARs at cerebellar synapses. AB - At many excitatory central synapses, activity produces a lasting change in the synaptic response by modifying postsynaptic AMPA receptors (AMPARs). Although much is known about proteins involved in the trafficking of Ca2+-impermeable (GluR2-containing) AMPARs, little is known about protein partners that regulate subunit trafficking and plasticity of Ca2+-permeable (GluR2-lacking) AMPARs. At cerebellar parallel fiber-stellate cell synapses, activity triggers a novel type of plasticity: Ca2+ influx through GluR2-lacking synaptic AMPARs drives incorporation of GluR2-containing AMPARs, generating rapid, lasting changes in excitatory postsynaptic current properties. Here we examine how glutamate receptor interacting protein (GRIP, also known as AMPAR binding protein or ABP) and protein interacting with C-kinase-1 (PICK) regulate subunit trafficking and plasticity. We find that repetitive synaptic activity triggers loss of synaptic GluR2-lacking AMPARs by selectively disrupting their interaction with GRIP and that PICK drives activity-dependent delivery of GluR2-containing receptors. This dynamic regulation of AMPARs provides a feedback mechanism for controlling Ca2+ permeability of synaptic receptors. PMID- 15895087 TI - RNA editing produces glycine receptor alpha3(P185L), resulting in high agonist potency. AB - The function of supramedullary glycine receptors (GlyRs) is still unclear. Using Wistar rat collicular slices, we demonstrate GlyR-mediated inhibition of spike discharge elicited by low glycine (10 microM). Searching for the molecular basis of this phenomenon, we identified a new GlyR isoform. GlyR alpha3(P185L), a result of cytidine 554 deamination, confers high glycine sensitivity (EC50 approximately 5 microM) to neurons and thereby promotes the generation of sustained chloride conductances associated with tonic inhibition. The level of GlyR alpha3-C554U RNA editing is sensitive to experimentally induced brain lesion, inhibition of cytidine deamination by zebularine and inhibition of mRNA transcription by actinomycin D, but not to blockade of protein synthesis by cycloheximide. Conditional regulation of GlyR alpha3(P185L) is thus likely to be part of a post-transcriptional adaptive mechanism in neurons with enhanced excitability. PMID- 15895088 TI - LINGO-1 negatively regulates myelination by oligodendrocytes. AB - The control of myelination by oligodendrocytes in the CNS is poorly understood. Here we show that LINGO-1 is an important negative regulator of this critical process. LINGO-1 is expressed in oligodendrocytes. Attenuation of its function by dominant-negative LINGO-1, LINGO-1 RNA-mediated interference (RNAi) or soluble human LINGO-1 (LINGO-1-Fc) leads to differentiation and increased myelination competence. Attenuation of LINGO-1 results in downregulation of RhoA activity, which has been implicated in oligodendrocyte differentiation. Conversely, overexpression of LINGO-1 leads to activation of RhoA and inhibition of oligodendrocyte differentiation and myelination. Treatment of oligodendrocyte and neuron cocultures with LINGO-1-Fc resulted in highly developed myelinated axons that have internodes and well-defined nodes of Ranvier. The contribution of LINGO 1 to myelination was verified in vivo through the analysis of LINGO-1 knockout mice. The ability to recapitulate CNS myelination in vitro using LINGO-1 antagonists and the in vivo effects seen in the LINGO-1 knockout indicate that LINGO-1 signaling may be critical for CNS myelination. PMID- 15895089 TI - CD14 is required for MyD88-independent LPS signaling. AB - The recessive mutation 'Heedless' (hdl) was detected in third-generation N-ethyl N-nitrosourea-mutated mice that showed defective responses to microbial inducers. Macrophages from Heedless homozygotes signaled by the MyD88-dependent pathway in response to rough lipopolysaccharide (LPS) and lipid A, but not in response to smooth LPS. In addition, the Heedless mutation prevented TRAM-TRIF-dependent signaling in response to all LPS chemotypes. Heedless also abolished macrophage responses to vesicular stomatitis virus and substantially inhibited responses to specific ligands for the Toll-like receptor 2 (TLR2)-TLR6 heterodimer. The Heedless phenotype was positionally ascribed to a premature stop codon in Cd14. Our data suggest that the TLR4-MD-2 complex distinguishes LPS chemotypes, but CD14 nullifies this distinction. Thus, the TLR4-MD-2 complex receptor can function in two separate modes: one in which full signaling occurs and one limited to MyD88-dependent signaling. PMID- 15895090 TI - Enhanced Toll-like receptor responses in the absence of signaling adaptor DAP12. AB - DAP12 is a signaling adaptor containing an immunoreceptor tyrosine-based activation motif (ITAM) that pairs with receptors on myeloid cells and natural killer cells. We examine here the responses of mice lacking DAP12 to stimulation through Toll-like receptors (TLRs). Unexpectedly, DAP12-deficient macrophages produced higher concentrations of inflammatory cytokines in response to a variety of pathogenic stimuli. Additionally, macrophages deficient in spleen tyrosine kinase (Syk), which signals downstream of DAP12, showed a phenotype identical to that of DAP12-deficient macrophages. DAP12-deficient mice were more susceptible to endotoxic shock and had enhanced resistance to infection by the intracellular bacterium Listeria monocytogenes. These data suggest that one or more DAP12 pairing receptors negatively regulate signaling through TLRs. PMID- 15895091 TI - Signaling conformations of the tall cytokine receptor gp130 when in complex with IL-6 and IL-6 receptor. AB - gp130 is a shared cytokine signaling receptor and the founding member of the 'tall' class of cytokine receptors. A crystal structure of the ligand-binding domains of gp130 in complex with human interleukin-6 (IL-6) and its a-receptor (IL-6Ralpha) revealed a hexameric architecture in which the gp130 membrane-distal regions were approximately 100 A apart, in contrast to the close apposition seen between short cytokine receptor complexes. Here we used single-particle EM to visualize the entire extracellular hexameric IL-6-IL-6Ralpha-gp130 complex, containing all six gp130 domains. The structure reveals that gp130 is bent such that the membrane-proximal domains of gp130 are close together at the cell surface, enabling activation of intracellular signaling. Variation in the receptor bend angles suggests a possible conformational transition from open to closed states upon ligand binding; this transition is probably representative of the other tall cytokine receptors. PMID- 15895092 TI - Insights into pneumococcal pathogenesis from the crystal structure of the modular teichoic acid phosphorylcholine esterase Pce. AB - Phosphorylcholine, a specific component of the pneumococcal cell wall, is crucial in pathogenesis. It directly binds to the human platelet-activating factor (PAF) receptor and acts as a docking station for the family of surface-located choline binding proteins (CBP). The first structure of a complete pneumococcal CBP, Pce (or CbpE), has been solved in complex with the reaction product and choline analogs. Pce has a novel modular structure, with a globular N-terminal module containing a binuclear Zn(2+) catalytic center, and an elongated choline-binding module. Residues involved in substrate binding and catalysis are described and modular configuration of the active center accounts for in vivo features of teichoic acid hydrolysis. The hydrolysis of PAF by Pce and its regulatory role in phosphorylcholine decoration of the bacterial surface provide new insights into the critical function of Pce in pneumococcal adherence and invasiveness. PMID- 15895093 TI - Solution structure of the HIV-1 integrase-binding domain in LEDGF/p75. AB - Lens epithelium-derived growth factor (LEDGF)/p75 is the dominant binding partner of HIV-1 integrase (IN) in human cells. We have determined the NMR structure of the integrase-binding domain (IBD) in LEDGF and identified amino acid residues essential for the interaction. The IBD is a compact right-handed bundle composed of five alpha-helices. Based on folding topology, the IBD is structurally related to a diverse family of alpha-helical proteins that includes eukaryotic translation initiation factor eIF4G and karyopherin-beta. LEDGF residues essential for the interaction with IN were localized to interhelical loop regions of the bundle structure. Interaction-defective IN mutants were previously shown to cripple replication although they retained catalytic function. The initial structure determination of a host cell factor that tightly binds to a retroviral enzyme lays the groundwork for understanding enzyme-host interactions important for viral replication. PMID- 15895094 TI - The RISC subunit Tudor-SN binds to hyper-edited double-stranded RNA and promotes its cleavage. AB - Long perfect double-stranded RNA (dsRNA) molecules play a role in various cellular pathways. dsRNA may undergo extensive covalent modification (hyper editing) by adenosine deaminases that act on RNA (ADARs), resulting in conversion of up to 50% of adenosine residues to inosine (I). Alternatively, dsRNA may trigger RNA interference (RNAi), resulting in silencing of the cognate mRNA. These two pathways have previously been shown to be antagonistic. We show a novel interaction between components of the ADAR and RNAi pathways. Tudor staphylococcal nuclease (Tudor-SN) is a subunit of the RNA-induced silencing complex, which is central to the mechanism of RNAi. Here we show that Tudor-SN specifically interacts with and promotes cleavage of model hyper-edited dsRNA substrates containing multiple I.U and U.I pairs. This interaction suggests a novel unsuspected interplay between the two pathways that is more complex than mutual antagonism. PMID- 15895095 TI - Biomaterials functionalization using a novel peptide that selectively binds to a conducting polymer. AB - The goal in biomaterial surface modification is to retain a material's bulk properties while modifying only its surface to possess desired recognition and specificity. Here we develop a unique strategy for surface functionalization of an electrically conductive polymer, chlorine-doped polypyrrole (PPyCl), which has been widely researched for various electronic and biomedical applications. An M13 bacteriophage library was used to screen 10(9) different 12-mer peptide inserts against PPyCl. A binding phage (phiT59) was isolated, and its binding stability and specificity to PPyCl was assessed using fluorescence microscopy and titer count analysis. The relative binding strength and mechanism of the corresponding 12-mer peptide and its variants was studied using atomic force microscopy and fluorescamine assays. Further, the T59 peptide was joined to a cell adhesive sequence and used to promote cell attachment on PPyCl. This strategy can be extended to immobilize a variety of molecules to PPyCl for numerous applications. In addition, phage display can be applied to other polymers to develop bioactive materials without altering their bulk properties. PMID- 15895096 TI - Inverse magnetocaloric effect in ferromagnetic Ni-Mn-Sn alloys. AB - The magnetocaloric effect (MCE) in paramagnetic materials has been widely used for attaining very low temperatures by applying a magnetic field isothermally and removing it adiabatically. The effect can also be exploited for room-temperature refrigeration by using giant MCE materials. Here we report on an inverse situation in Ni-Mn-Sn alloys, whereby applying a magnetic field adiabatically, rather than removing it, causes the sample to cool. This has been known to occur in some intermetallic compounds, for which a moderate entropy increase can be induced when a field is applied, thus giving rise to an inverse magnetocaloric effect. However, the entropy change found for some ferromagnetic Ni-Mn-Sn alloys is just as large as that reported for giant MCE materials, but with opposite sign. The giant inverse MCE has its origin in a martensitic phase transformation that modifies the magnetic exchange interactions through the change in the lattice parameters. PMID- 15895097 TI - Non-viral gene delivery regulated by stiffness of cell adhesion substrates. AB - Non-viral gene vectors are commonly used for gene therapy owing to safety concerns with viral vectors. However, non-viral vectors are plagued by low levels of gene transfection and cellular expression. Current efforts to improve the efficiency of non-viral gene delivery are focused on manipulations of the delivery vector, whereas the influence of the cellular environment in DNA uptake is often ignored. The mechanical properties (for example, rigidity) of the substrate to which a cell adheres have been found to mediate many aspects of cell function including proliferation, migration and differentiation, and this suggests that the mechanics of the adhesion substrate may regulate a cell's ability to uptake exogeneous signalling molecules. In this report, we present a critical role for the rigidity of the cell adhesion substrate on the level of gene transfer and expression. The mechanism relates to material control over cell proliferation, and was investigated using a fluorescent resonance energy transfer (FRET) technique. This study provides a new material-based control point for non viral gene therapy. PMID- 15895098 TI - Shape-memory nanoparticles from inherently non-spherical polymer colloids. AB - Samples of polymeric materials generally have no intrinsic shape; rather their macroscopic form is determined by external forces such as surface tension and memory of shear (for example, during extrusion, moulding or embossing). Hence, in the molten state, the thermodynamically most stable form for polymer (nano)particles is spherical. Here, we present the first example of polymer nanoparticles that have an intrinsic non-spherical shape. We observe the formation of high-aspect-ratio ellipsoidal polymer nanoparticles, of controlled diameter, made from main-chain liquid crystalline polymers using a mini-emulsion technique. The ellipsoidal shape is shown to be an equilibrium (reversible) characteristic and a direct result of the material shape memory when a liquid crystal nanoparticle is in its monodomain form. PMID- 15895099 TI - Pressure-induced ferromagnetism in (In,Mn)Sb dilute magnetic semiconductor. AB - Recent advances in III(1-x)Mn(x)V ferromagnetic semiconductors (for example in Ga(1-x)Mn(x)As) have demonstrated that electrical control of their spin properties can be used for manipulation and detection of magnetic signals. The Mn(2+) ions in these alloys provide magnetic moments, and at the same time act as a source of valence-band holes that mediate the Mn(2+)-Mn(2+) interactions. This coupling results in the ferromagnetic phase. In earlier workit was shown that the ferromagnetic state can be enhanced or suppressed by varying the carrier density. Here we demonstrate that, by using hydrostatic pressure to continuously tune the wavefunction overlap, one can control the strength of ferromagnetic coupling without any change in the carrier concentration. Tuning the exchange coupling by this process increases the magnetization spectacularly, and can even induce the ferromagnetic phase in an initially paramagnetic alloy. These results may open new directions for strain-engineering of nanodevices. PMID- 15895100 TI - Nanowire dye-sensitized solar cells. AB - Excitonic solar cells-including organic, hybrid organic-inorganic and dye sensitized cells (DSCs)-are promising devices for inexpensive, large-scale solar energy conversion. The DSC is currently the most efficient and stable excitonic photocell. Central to this device is a thick nanoparticle film that provides a large surface area for the adsorption of light-harvesting molecules. However, nanoparticle DSCs rely on trap-limited diffusion for electron transport, a slow mechanism that can limit device efficiency, especially at longer wavelengths. Here we introduce a version of the dye-sensitized cell in which the traditional nanoparticle film is replaced by a dense array of oriented, crystalline ZnO nanowires. The nanowire anode is synthesized by mild aqueous chemistry and features a surface area up to one-fifth as large as a nanoparticle cell. The direct electrical pathways provided by the nanowires ensure the rapid collection of carriers generated throughout the device, and a full Sun efficiency of 1.5% is demonstrated, limited primarily by the surface area of the nanowire array. PMID- 15895101 TI - Metabolic and vascular actions of endothelin-1 are inhibited by insulin-mediated vasodilation in perfused rat hindlimb muscle. AB - Endothelin-1 (ET-1) is a potent endothelium-derived vasoactive peptide and may be involved in the microvascular actions of insulin for the normal delivery of nutrients to muscle, although higher levels may be antagonistic. Our aim was to observe the interaction between ET-1 and insulin. Initially, we attempted to distinguish the vascular from the metabolic effects of ET-1 in the constant-flow pump-perfused rat hindlimb by using various doses of ET-1 and measuring changes in perfusion pressure (PP), oxygen consumption (VO(2)), glucose uptake (GU) and lactate release (LR). Sodium nitroprusside (SNP) was used to block vasoconstriction and to thus assess the relationship between vascular and metabolic effects. Insulin was included in later experiments to determine the interaction between insulin and ET-1 on the above parameters. ET-1 caused a dose dependent increase in PP. Effects on VO(2) were biphasic, with low doses increasing VO(2), and higher doses leading to a net inhibition. GU and LR were increased at lower doses (ET-1 < or =1 nM), but this effect was lost at higher doses (> or =10 nM ET-1). SNP (50 microM) fully blocked the increase in pressure and metabolism due to low-dose ET-1 and partly blocked both pressure and metabolic responses by the high dose. ET-1 vasodilatory activity was minimal at high or low dose. Insulin (15 nM) alone caused GU, which was not affected by ET 1. Of the other parameters measured, insulin behaved essentially the same as SNP, inhibiting the pressure and oxygen effects. Overall, these results show that ET-1 has a biphasic dose-dependent vasoconstrictor effect on hindlimb blood vessels, able to modulate flow to cause both the stimulation and inhibition of metabolism, although these effects are blocked by insulin, which is able to vasodilate against both low and high doses of ET-1. PMID- 15895102 TI - New insights into adenosine-mediated myocardial protection. PMID- 15895103 TI - Cardioprotection with adenosine: 'a riddle wrapped in a mystery'. AB - Review of the published literature on adenosine and cardioprotection could lead one to paraphrase the famous words of Sir Winston Churchill (Radio broadcast, 1 October 1939 (in reference to Russia)) and conclude: 'I cannot forecast to you the action of adenosine. It is a riddle wrapped in a mystery inside an enigma'. That is, although it is well-established that adenosine can render cardiomyocytes resistant to lethal ischemia/reperfusion-induced injury, new and intriguing insights continue to emerge as to the mechanisms by which adenosine might limit myocardial infarct size. PMID- 15895104 TI - Intravenous adenosine protects the myocardium primarily by activation of a neurogenic pathway. AB - Endogenous adenosine is a trigger for ischemic myocardial preconditioning (IPC). Although intravascular administration of adenosine has been used to further unravel the mechanism of protection by IPC, it is questionable whether adenosine and IPC employ the same signaling pathways to exert cardioprotection. We therefore investigated whether the active metabolic barrier of the endothelium prevents an increase in myocardial interstitial adenosine concentrations by intravenous adenosine, using microdialysis, and also the role of NO and activation of a neurogenic pathway in the cardioprotection by adenosine. In pentobarbital-anesthetized rats, area at risk and infarct size (IS) were determined 120 min after a 60-min coronary artery occlusion (CAO), using trypan blue and nitro-blue-tetrazolium staining, respectively. IPC with a single 15-min CAO and a 15-min adenosine infusion (ADO, 200 microg min(-1) i.v.) limited IS to the same extent (IS = 41 +/- 6% and IS = 40 +/- 4%, respectively) compared to control rats (IS = 63 +/- 3%, both P < 0.05). However, IPC increased myocardial interstitial adenosine levels seven-fold from 4.3 +/- 0.7 to 27.1 +/- 10.0 microM (P < 0.05), while ADO had no effect on interstitial adenosine (4.1 +/- 1.2 microM), or any of the other purines. The NO synthase inhibitor N(omega)-nitro-L arginine (LNNA), which did not affect IS (IS = 62 +/- 3%), attenuated the protection by ADO (IS = 56 +/- 3%; P < 0.05 vs ADO, P = NS vs LNNA). The ganglion blocker hexamethonium, which had also no effect on IS (IS = 66 +/- 3%), blunted the protection by ADO (IS = 55 +/- 4%; P < 0.05 vs ADO and vs hexamethonium). These observations demonstrate that cardioprotection by ADO is dependent on NO, and is primarily mediated by activation of a neurogenic pathway. PMID- 15895105 TI - Bradykinin-induced, endothelium-dependent responses in porcine coronary arteries: involvement of potassium channel activation and epoxyeicosatrienoic acids. AB - In coronary arteries, bradykinin opens endothelial intermediate- and small conductance Ca2+-sensitive K+ channels (IK(Ca) and SK(Ca)) and, additionally, releases epoxyeicosatrienoic acids (EETs) from the endothelium. To clarify the involvement of these pathways in endothelium-dependent myocyte hyperpolarization, bradykinin-induced electrical changes in endothelial cells and myocytes of porcine coronary arteries (following nitric oxide (NO) synthase and cyclooxygenase inhibition) were measured using sharp microelectrodes. Hyperpolarization of endothelial cells by bradykinin (27.0 +/- 0.9 mV, n = 4) was partially inhibited (74%) by blockade of IK(Ca) and SK(Ca) channels using 10 microM TRAM-39 (2-(2-chlorophenyl)-2,2-diphenylacetonitrile) plus 100 nM apamin (leaving an iberiotoxin-sensitive component), whereas the response to substance P was abolished. After gap junction blockade with HEPES, (N-(2 hydroxyethyl)piperazine-N'-(2-ethanesulphonic acid)) hyperpolarization of the endothelium by 100 nM bradykinin was abolished by TRAM-39 plus apamin, whereas myocyte hyperpolarization still occurred (12.9 +/- 1.0 mV, n=4). The residual hyperpolarizations to 100 nM bradykinin were antagonized by the EET antagonist, 14,15-EEZE (14,15-epoxyeicosa-5(Z)-enoic acid) (10 microM), and abolished by iberiotoxin. Bradykinin-induced myocyte hyperpolarizations were also reduced by 14,15-EEZE-mSI (14,15-EEZE-methylsulfonylimide) (5,6- and 14,15-EET antagonist), whereas those to exogenous 11,12-EET were unaffected. These data show that bradykinin-induced hyperpolarization of endothelial cells (due to the opening of IK(Ca) and SK(Ca) channels) is electrotonically transferred to the myocytes via gap junctions. Bradykinin (but not substance P) also hyperpolarizes myocytes by a mechanism (independent of endothelial cell hyperpolarization) which involves endothelial cell production of EETs (most likely 14,15- and/or 11,12-EET). These open endothelial IK(Ca) and SK(Ca) channels and also activate large-conductance calcium-sensitive K+ channels (BK(Ca)) on the surrounding myocytes. PMID- 15895106 TI - Heteroactivation of cytochrome P450 1A1 by teas and tea polyphenols. AB - We studied 7-ethoxyresorufin deethylase as an index of cytochrome P4501A1 (CYP1A1) activity in liver microsomes from rats pretreated with 3 methylcholanthrene. The enzyme had complex kinetics compatible with a multisite model. At 1 microM substrate, brewed black, green and white teas had complex effects on enzyme activity consisting of activation at low concentrations and inhibition at higher concentrations. Data fit well to a two-site model that allowed us to determine maximal activation (% increase above control), pEC(50) for activation (g ml(-1)) and pIC(50) for inhibition (g ml(-1)). These parameters were 190+/-40, 5.9+/-0.1 and 4.51+/-0.09 for green tea, 350+/-40, 5.43+/-0.05 and 5.43+/-0.05 for black tea and 230+/-80, 5.3+/-0.3 and 4.7+/-0.2 for white tea, respectively. The effects of the brewed teas were mimicked to different degrees by the green tea polyphenols. Maximal activation, pEC(50) (M) and pIC(50) (M) were: (-)-epicatechin, 55+/-9, 5.4+/-0.3, 2+/-1; (-)-epicatechin gallate, 160+/ 60, 6.2+/-0.3, 5.28+/-0.06; (-)-epigallocatechin 30+/-10, 6.5+/-0.5, 3.37+/-0.08; and (-)-epigallocatechin gallate 130+/-40, 6.7+/-0.3, 5.0+/-0.1. A crude extract of black tea polyphenols inhibited 7-ethoxyresorufin deethylase, but did not cause enzyme activation consistently. Enzyme activation was dependent upon substrate concentration. Heteroactivation of CYP1A1 may partially explain the lack of agreement between biological and epidemiological evidence of a role for tea in cancer prevention. PMID- 15895107 TI - Inhibition of fatty acid amide hydrolase and monoacylglycerol lipase by the anandamide uptake inhibitor VDM11: evidence that VDM11 acts as an FAAH substrate. AB - There is some dispute concerning the extent to which the uptake inhibitor VDM11 (N-(4-hydroxy-2-methylphenyl) arachidonoyl amide) is capable of inhibiting the metabolism of the endocannabinoid anandamide (AEA) by fatty acid amide hydrolase (FAAH). In view of a recent study demonstrating that the closely related compound AM404 (N-(4-hydroxyphenyl)arachidonylamide) is a substrate for FAAH, we re examined the interaction of VDM11 with FAAH. In the presence of fatty acid-free bovine serum albumin (BSA, 0.125% w v(-1)), both AM404 and VDM11 inhibited the metabolism of AEA by rat brain FAAH with similar potencies (IC(50) values of 2.1 and 2.6 microM, respectively). The compounds were about 10-fold less potent as inhibitors of the metabolism of 2-oleoylglycerol (2-OG) by cytosolic monoacylglycerol lipase (MAGL). The potency of VDM11 towards FAAH was dependent upon the assay concentration of fatty acid-free bovine serum albumin (BSA). Thus, in the absence of fatty acid-free BSA, the IC(50) value for inhibition of FAAH was reduced by a factor of about two (from 2.9 to 1.6 microM). A similar reduction in the IC(50) value for the inhibition of membrane bound MAGL by both this compound (from 14 to 6 microM) and by arachidonoyl serinol (from 24 to 13 microM) was seen. An HPLC assay was set up to measure 4-amino-m-cresol, the hypothesised product of FAAH-catalysed VDM11 hydrolysis. 4-Amino-m-cresol was eluted with a retention time of approximately 2.4 min, but showed a time dependent degradation to compounds eluting at peaks of approximately 5.6 and approximately 8 min. Peaks with the same retention times were also found following incubation of the membranes with VDM11, but were not seen when the membranes were preincubated with the FAAH inhibitors URB597 (3'-carbamoyl biphenyl-3-yl-cyclohexylcarbamate) and CAY10401 (1-oxazolo[4,5-b]pyridin-2-yl-9 octadecyn-1-one) prior to addition of VDM11. The rate of metabolism of VDM11 was estimated to be roughly 15-20% of that for anandamide. It is concluded that VDM11 is an inhibitor of FAAH under the assay conditions used here, and that the inhibition may at least in part be a consequence of the compound acting as an alternative substrate. PMID- 15895108 TI - Lipids modulate ligand binding to sulphonylurea receptors. AB - ATP-sensitive K(+) channels (K(ATP) channels) are complexes of inwardly rectifying K(+) channels (Kir6.x) and sulphonylurea receptors (SURs). Kir6.2 containing channels are closed by ATP binding to Kir6.2, and opened by MgADP binding to SUR. Channel activity is modulated by synthetic compounds such as the channel-blocking sulphonylureas and the K(ATP) channel openers, which both act by binding to SUR. By interacting with Kir6.2, phosphatidylinositol-4,5-bisphosphate (PIP(2)) and oleoyl-coenzyme A (OCoA) decrease the ATP-sensitivity of the channel and abolish the effect of the synthetic channel modulators. Here, we have investigated whether lipids and related compounds interfered with the binding of the sulphonylurea, glibenclamide (GBC) and of the opener, N-cyano-N'-(1,1 dimethylpropyl)-N''-3-pyridylguanidine (P1075), to the SUR subtypes. Lipids (100 300 microM) inhibited binding of [(3)H]GBC and [(3)H]P1075 to SUR subtypes in the rank order OCoA>dioleylglycerol-succinyl-nitriloacetic acid (DOGS NTA)>oleate>malonyl-CoA>PIP(2.)OCoA inhibited radioligand binding to SUR completely, with IC(50) values ranging from 6 to 44 microM. Inhibition was reversed by increasing the concentration of the radioligands in agreement with an essentially competitive mechanism. MgATP and coexpression with Kir6.2 decreased the potency of OCoA. DOGS-NTA inhibited radioligand binding to SUR by 40-88%, with IC(50) values ranging from 38 to 120 microM. Poly-lysine increased radioligand binding to SUR by up to 30% but did not affect much the inhibition of ligand binding by OCoA and DOGS-NTA. Radioligand binding to SUR2A but not to the other SUR subtypes was slightly (10-20%) stimulated by lipids at concentrations approximately 10 x lower than required for inhibition. The data show that certain lipids, at high concentrations, interact with SUR and inhibit the binding of GBC and P1075; with SUR2A, a modest stimulation of ligand binding precedes inhibition. Regarding K(ATP) channel activity, these effects will be overruled by the interaction of the lipids with Kir6.2 at lower (physiological) concentrations. They are, however, of pharmacological importance and must be taken into account if high concentrations of these compounds (e.g. OCoA>10 microM) are used to interfere with the action of sulphonylureas and openers on K(ATP) channel activity. PMID- 15895109 TI - The ventral tegmental area as a putative target for tachykinins in cardiovascular regulation. AB - Tachykinin receptor agonists and antagonists were microinjected into the ventral tegmental area (VTA) to study the relative participation of the three tachykinin receptors in cardiovascular regulation in freely behaving rat. Selective agonists (1-100 pmol) for NK1 ([Sar9, Met (O2)11]SP), NK2 ([beta-Ala8]NKA (4-10)) and NK3 (senktide) receptors evoked increases in blood pressure, heart rate (HR) along with behavioural manifestations (face washing, sniffing, head scratching, rearing, wet dog shake). At 1 pmol, NK1 and NK3 agonists did not affect behaviour and blood pressure but only HR. Tachykinin agonists-induced cardiovascular responses were selectively and reversibly blocked by the prior injection of antagonists for NK1 receptors (LY 303870 ((R)-1-[N-(2-methoxybenzyl)acetylamino] 3-(1H-indol-3-yl)-2-[N-(2-(4-(piperidin-1-yl)piperidin-1 yl)acetyl)amino]propane), 5 nmol), NK2 receptors (SR 48968 ([(S)-N-methyl-N-[4 acetylamino-4-phenylpiperidino-2-(3,4-dichlorophenyl)butyl]benzamide]), 250 pmol) and NK3 receptors (SB 235375 ((-)-(S)-N-(alpha-ethylbenzyl)-3-(carboxymethoxy)-2 phenylquinoline-4-carboxamide), 25 nmol). With the exception of the NK2 agonist, most behavioural effects were also blocked by antagonists. Tachykinin agonists induced cardiovascular responses were inhibited by intravenous (i.v.) treatments with antagonists for D1 dopamine receptor (SCH23390, 0.2 mg kg(-1)) and beta1 adrenoceptor (atenolol, 5 mg kg(-1)) but not for D2 dopamine receptor (raclopride, 0.16 mg kg(-1)). Behavioural responses were blocked by SCH23390 only. The present study provides the first pharmacological evidence that the three tachykinin receptors in the rat VTA can affect the autonomic control of blood pressure and HR by increasing midbrain dopaminergic transmission. This mechanism may be involved in the coordination of behavioural and cardiovascular responses to stress and noxious stimulation. PMID- 15895110 TI - Key roles of hydrophobic rings of TM2 in gating of the alpha9alpha10 nicotinic cholinergic receptor. AB - We have performed a systematic mutagenesis of three hydrophobic rings (17', 13' and 9') within transmembrane region (TM) 2 of the alpha9alpha10 nicotinic cholinergic receptor (nAChR) to a hydrophilic (threonine) residue and compared the properties of mutant receptors reconstituted in Xenopus laevis oocytes. Phenotypic changes in alpha9alpha10 mutant receptors were evidenced by a decrease in the desensitization rate, an increase in both the EC(50) for ACh as well as the efficacy of partial agonists and the reduction of the allosteric modulation by extracellular Ca(2+). Mutated receptors exhibited spontaneous openings and, at the single-channel level, an increased apparent mean open time with no major changes in channel conductance, thus suggesting an increase in gating of the channel as the underlying mechanism. Overall, the degrees of the phenotypes of mutant receptors were more overt in the case of the centrally located V13'T mutant. Based on the atomic model of the pore of the electric organ of the Torpedo ray, we can propose that the interactions of side chains at positions 13' and 9' are key ones in creating an energetic barrier to ion permeation. In spite of the fact that the roles of the TM2 residues are mostly conserved in the distant alpha9alpha10 member of the nAChR family, their mechanistic contributions to channel gating show significant differences when compared to other nAChRs. These differences might be originated from slight differential intramolecular rearrangements during gating for the different receptors and might lead each nAChR to be in tune with their physiological roles. PMID- 15895111 TI - Antidiabetic sulphonylureas activate mitochondrial permeability transition in rat skeletal muscle. AB - Antidiabetic sulphonylureas can bind to various intracellular organelles including mitochondria. The aim of this study was to monitor the influence of antidiabetic sulphonylureas on membrane permeability in mitochondria isolated from rat skeletal muscle. The effects of glibenclamide (and other sulphonylurea derivatives) on mitochondrial function were studied by measuring mitochondrial swelling, mitochondrial membrane potential, respiration rate and Ca2+ transport into mitochondria. We observed that glibenclamide induced mitochondrial swelling (EC50 = 8.2 +/- 2.5 microM), decreased the mitochondrial membrane potential and evoked Ca2+ efflux from the mitochondrial matrix. These effects were blocked by 2 microM cyclosporin A, an inhibitor of the mitochondrial permeability transition. Moreover, 30 microM glibenclamide accelerated the respiratory rate in the presence of glutamate/malate, substrates of complex I of the mitochondrial respiratory chain. In conclusion, we postulate that the antidiabetic sulphonylureas activate the mitochondrial permeability transition in skeletal muscle by increasing its sensitivity to Ca2+. PMID- 15895112 TI - The graft-versus-lymphoma effect: clinical review and future opportunities. AB - Numerous lines of preclinical and clinical evidence support the existence of a graft-versus-leukemia effect, but less evidence supporting a comparable graft versus-lymphoma effect exists. We review here current clinical data addressing the graft-versus-lymphoma effect, including comparisons of autologous, syngeneic, and allogeneic transplantation; responses to immunomodulation; and responses to nonmyeloablative stem cell transplantation. Despite several limitations of the data, we believe that there is sufficient evidence suggesting a significant graft versus-lymphoma effect. In addition, we discuss approaches for clinical management of lymphoma patients, opportunities for mechanistic studies afforded by donor leukocyte infusions and nonmyeloablative transplantation, and suggestions for clinical studies to further define the magnitude and applicability of the graft-versus-lymphoma effect. PMID- 15895113 TI - Complete remission after myeloablation with bone-seeking 186Re-HEDP and high-dose melphalan followed by autologous stem cell transplantation in a patient with chemorefractory multiple myeloma. PMID- 15895114 TI - Expansion of cord blood CD34+ hematopoietic progenitor cells in coculture with autologous umbilical vein endothelial cells (HUVEC) is superior to cytokine supplemented liquid culture. AB - Expansion of hematopoietic progenitor cells (HPC) in the presence of endothelium has been shown to result in grafts capable of restoring hematopoiesis in a myeloablated host. However, the use of xenogeneic endothelium or cell lines may carry risks in a clinical transplantation setting. We explored the feasibility of cord blood progenitor cell expansion in vitro in an autologous coculture system using umbilical vein endothelial cells (HUVEC). CD34+ HPC and HUVEC were isolated from the same umbilical cord. For 3 days, HPC were maintained in serum-free medium supplemented with a single cytokine (SCF) or a cytokine combination (SCF, Flt3-ligand, IL-6). Meanwhile, adherent HUVEC cultures were established. After addition of VEGF and IL-1 at day 3, the cells were either added to HUVEC cultures or grown without endothelial cells for further 7 days. Total cells, CD34+ and clonogenic progenitors were significantly increased when coculture was compared to liquid culture. Long-term culture-initiating cells (LTC-IC) and cobble stone area-forming cells (CAFC, limiting dilution analysis) were detected more frequently after coculture with endothelial cells. Also precursors and mature myeloid cells were observed after expansion. We conclude that coculture with autologous HUVEC represents a feasable approach for ex vivo expansion of cord blood HPC. PMID- 15895115 TI - An elective single autograft with high-dose melphalan: single-center study of 451 patients. AB - In all, 451 myeloma patients, 51% previously untreated, underwent elective single autotransplantation after 200 mg/m(2) melphalan between 1985 and 2001 at the Royal Marsden Hospital. The therapy sequence was: Induction (vincristine, doxorubicin, methylprednisolone+/-cyclophosphamide), marrow or filgrastim mobilized blood stem cell harvest, autograft, and interferon-alpha2b maintenance. A total of 27 (6%) died of transplant-related toxicity, all within 3 months. Complete or near-complete remission was seen in 59% with an overall response rate of 91%. Subsequent disease progression was seen in 285, and 17 died of unrelated causes. In all, 206 patients were alive at the last follow-up, 6 months to 17.7 years post-transplant (median 65 months); 122 without disease progression at 6 months to 17.7 years (median 58 months). The median overall (OS) and event-free (EFS) survivals were 5.9 and 2.4 years, with 10-year OS and EFS probabilities of 31.4 and 16.5%, respectively. In Cox analysis, it was seen that significantly longer OS occurred for patients who had beta-2-microglobulin <3.5 mg/l (P<0.0001), age <60 years (P=0.001) and albumin > or =35 g/l (P=0.009). EFS was also longer if beta-2-microglobulin was <3.5 mg/l (P=0.0056) and patients were <60 years of age (P=0.033). We conclude that with a single planned autograft, patients with myeloma have an excellent outcome. PMID- 15895116 TI - Treatment of cutaneous chronic graft-versus-host disease with topical pimecrolimus. PMID- 15895117 TI - Reduced intensity conditioning bone marrow transplantation for pure red cell aplasia: successful outcome but difficult post transplant course. PMID- 15895120 TI - [Surgical performance]. PMID- 15895121 TI - [Isolated surgery of tricuspid insufficiency in valvular reoperation]. AB - BACKGROUND AND AIMS OF THE STUDY: Development of late tricuspid regurgitation (TR) following prior operation on left heart valves is associated with an important impairement of functional status and adverse prognosis. Indications and outcomes of surgery in this setting are not well defined. Our Department experience in isolated TR in valve reoperation is evaluated. METHODS: Twenty-four patients underwent isolated tricuspid valve surgery because of severe TR following a prior operation on left heart valves between January/1990 and July/2003. Patients with congenital heart disease, traumatic TR, infectious endocarditis or left heart valve disfunction were excluded. The retrospective and observational study of this group of patients included clinical echocardiographic, haemodynamic and surgical data evaluation. RESULTS: Mean follow-up time was 4.4+/-3.5 years and included all patients. There was no operative mortality but actuarial survival at six years was 48%. No risk factors for late death were identified. CONCLUSIONS: Isolated tricuspid surgery for severe TR in patients with previous left heart valve surgery can be done with acceptable hospital mortality. Low operative mortality does not mean medium term good results. High late mortality and unsatisfactory functional results should lead us to consider other options when treating tricuspid valve regurgitation. PMID- 15895122 TI - [Coronary revascularization with extracorporeal circulation, aortic clamping or OPCAB: does the method's choice influence the results?]. AB - BACKGROUND AND OBJECTIVES: Off pump CABG-OPCAB is the most recent development in coronary surgery and is aimed to avoid the complications of extracorporeal circulation and to compete with interventional cardiology. The objective of this study is to compare 3 methods for cardiac support for CABG. METHODS: Between February 2001 and December 2002 ninety consecutive patients were operated by the same surgeon and the same team with the following techniques: extracorporeal circulation (CEC) and ischaemic arrest, CEC and beating heart, and OPCAB. RESULTS: There were no difference in mortality and morbidity for the 3 groups of patients. The OPCAB group showed less ventilation time, less blood transfusion needs and shorter hospital stays than the other 2 groups of patients that showed no differences between each other. CONCLUSION: OPCAB is safe and showed slight advantage over CEC supported groups but the number of grafts per patient was smaller. PMID- 15895123 TI - [Pulmonary carcinoid tumors--ten years experience]. AB - Pulmonary carcinoid tumors are rare, accounting for as many as 2% of all pulmonary neoplasms and for 10% of carcinoid tumors overall. Previously classified as bronchial adenomas, actually are classified as neuroendocrine tumors. They have a subclassification into typical classed as low-grade malignant neoplasm and atypical more aggressive, with more potential to cause local invasion. In this paper, the authors report a retrospective study of 25 patients, who had the diagnosis of pulmonary carcinoid tumors and had been operated between January of 1994 and August of 2004. We conclude that this tumors must be considered malignant in the surgical approach. PMID- 15895124 TI - [Techniques and "tricks": different uses of Foley catheter in cardiovascular surgery]. AB - The use of Foley catheter is well established for bladder catheterization. The authors have used it, occasionally, in many unconventional ways to cope with difficult situations found at surgery. 1.Catastrophic haemorrhage during redo sternotomy. 2. Non touch technique for graft to aorta proximal anastomosis during CABG surgery in patients with heavily calcified aortas. 3. Proximal anastomosis of ventricle to pulmonary artery conduits, done with a beating heart. The use of this simple ''Colombo egg'' method has been useful to us in many difficult situations. It is safe and non expensive and should make part of everyone's surgical armamentarium. PMID- 15895125 TI - [Radiation-induced lesions of carotid arteries: report of five cases]. AB - This paper is aimed at the analysis and discussion of a personal experience with an unusual entity, the radiation-induced lesions of carotid arteries, based on 5 patients that were observed for the last twelve years. The series is composed of 4 men and woman, with an average age of 65 (between 51 to 77). Malignancies that required radiotherapy were diverse and the carotid lesions were recognized between 8 and 20 years after the irradiation (average 15) . They consisted in 3 occlusions and 5 stenosis of the internal carotid, 4 occlusions of the external carotid, and 2 stenosis of the common carotid artery. Four patients presented with symptoms of cerebrovascular insufficiency and 1 was asymptomatic. Three patients underwent 4 surgical procedures (1 bilateral) which consisted uniformly in the segmental resection of the stenosed arteries followed by its replacement with a graft, autologous or prosthetic. There was no operative mortality, nor significant morbidity. The main features of etiopathogeny, diagnosis and treatment of this entity are discussed, based on data published in literature, where a controversy between conventional surgery and endovascular management is evident. Conventional surgery is recognized as an effective method, bearing excellent and durable results. Endovascular management requires more consistent and long-lasting results, to be confirmed and assumed as a reliable alternative to the surgical treatment of this peculiar form of carotid arteries pathology. PMID- 15895126 TI - [Aneurysm of the superior mesenteric artery in the context of a multianeurismatic disease. Case report]. AB - Aneurysms of the superior mesenteric artery are one of the most uncommon visceral artery aneurysms. Despite its rarity they course with high risks of rupture and many constitute a permanent threat to patients's life. The authors report the clinical condition of a male patient, with a large aneurysm of the superior mesenteric artery, diagnosed through the image techniques made during the follow up of a previously operated aorto-iliac and bilateral popliteal aneurysms. Its clinical presentation and the surgical management are subjected to presentation and discussion. PMID- 15895127 TI - [Phlegmasia caerulea dolens: therapeutic considerations]. AB - Phlegmasia caerulea dolens is a rare complication of a deep venous thrombosis and is the result of a massive occlusion of all venous outflow of the extremity. Diagnosis must be made early in the course of the process for treatment effectiveness although it can only produce modest results. The authors analyse retrospectively 3 cases that were treated between 2001 and 2005. The three patients suffered from malignancies (2 from lung and one fom prostate). In all patients a venous thrombectomy was performed and the method employed was described, which includes the systematic use of a caval filter. Two patients coursed with good results, with complete resolution or minor amputation. However, one patient needed an above knee amputation. The authors concluded that the time that mediate to the precise diagnosis and the best option of treatment are important determinations on the results, that however are very dependent on the etiology of this clinical situation. PMID- 15895128 TI - [Spinal cord ischemia after thoracic and thoracoabdominal aortic aneurysm repair]. AB - In this paper, the author reviews the problematic around the spinal cord ischemic disfunction that sometimes occurs following the surgical management of thoracic and thoracoabdominal aortic aneurysm. After an anatomical review of the spinal cord vascularization, the diverse pathogenic mechanisms involved are described together with its importance and clinical significance, as well as the multiple procedures, techniques and pharmacotherapy employed nowadays aimed at lowering the occurrence of this most dramatic complication of thoracic and thoracoabdominal aortic aneurysm repair. PMID- 15895129 TI - [Rupture of pulsatile varices in the elderly]. PMID- 15895130 TI - [Influence of grandmothers on breastfeeding practices]. AB - OBJECTIVE: To assess the influence of grandmothers on breastfeeding practices. METHODS: This was a prospective study on 601 mothers of normal babies born in a university hospital in the city of Porto Alegre, State of Rio Grande do Sul. Data were collected in the maternity ward and at home, one, two, four and six months after delivery, by means of interviews with the mothers. Information about grandmothers was obtained at the time of the first home visit. Multiple logistic regression was used to test associations between variables related to the grandmothers and the prevalence of breastfeeding. RESULTS: Abandonment of exclusive breastfeeding within the first month was significantly associated with maternal or paternal grandmothers who advised that water or tea (OR=2.2 and 1.8, respectively) and other kinds of milk (OR=4.5 and 1.9, respectively) should be given. Abandonment of breastfeeding within the first six months was associated with maternal and paternal grandmothers who advised that other kinds of milk (OR=2.4 and 2.1, respectively) should be given. Non-daily contact with the maternal grandmother was a protective factor for maintaining breastfeeding until six months. CONCLUSIONS: Grandmothers may have a negative influence on breastfeeding, both on its duration and its exclusivity. This information could be useful in the planning of strategies for promoting breastfeeding. PMID- 15895131 TI - [Assessment of lunch served in the Workers' Food Program, Brazil]. AB - OBJECTIVE: In the light of the Workers' Food Program (WFP) growth and its recent review of nutritional parameters regulations, the study aimed at evaluating food intake in WFP through dietary assessment of lunch served in the program and workers' nutritional status. METHODS: A cross-sectional study was carried out in a representative sample of workers in Brasilia, Federal District, Brazil. A total of 1,044 subjects who had lunch at 52 food and nutrition units were evaluated. Social-economic and demographic data were collected as well as anthropometric measures for calculating the Body Mass Index. Food intake was assessed by dish weight and direct observation of dish composition. RESULTS: Of all subjects, 43% had excess weight, 33.7% were overweight and 9.3% were obese. Males were most affected. Median lunch energy intake was 515 kcal in women and 736 kcal in men. Median dietary fiber intake was 6.0 g among women and 8.3 g among men, and median cholesterol intake was over 90 mg among subjects with excess weight. CONCLUSIONS: The results indicate that the study population who is often seen as healthy is at nutritional risk. Workers in WFP should be targeted for health promotion strategies using especially nutritionists' skills as educators. PMID- 15895132 TI - [The symbolic and utilitarian facets of pacifiers according to mothers]. AB - OBJECTIVE: Despite governmental initiatives that control, discourage and even prohibit the divulgation of the usage of dummies and pacifiers in maternity wards, the frequency of pacifier use by Brazilian children is still high. In light of this phenomenon, the aim of the present study was to investigate the social representations of the pacifier constructed by mothers whose children used pacifiers. METHODS: The present study is based on the conceptual framework of social representations, proposed by Moscovici. We studied women who gave birth at a teaching hospital in the city of Sao Paulo that prohibits the use of pacifiers during hospital admission. We conducted non-structured individual interviews that were transcribed in full and organized for analysis according to the collective subject discourse method. RESULTS: Our results indicate maternal representations that the pacifier "symbolizes the child," that is "is a tranquilizer for the child and an aid for the mother," and that "its use is inherited from generation to generation". CONCLUSIONS: Pacifiers are an alternative for the mother for comforting and hushing her child in moments of agitation or for when the mother is not able to tend to the child in a direct and continuous manner. PMID- 15895133 TI - [Body mass index and measures of adiposity among elderly adults]. AB - OBJECTIVE: To evaluate the nutritional status of elderly persons and to compare the correlation between the body mass index (BMI=kg/m2) with measures of adiposity and fat distribution among elderly and middle-aged adults. METHODS: Elderly persons (N=699; 60 years or older) and middle-aged adults (N=1,306; 40 59.9 years) participating in a population-based survey conducted in 1996 in the municipality of Rio de Janeiro were evaluated as to body mass index, arm, waist, and hip circumferences, skinfolds, and arm fat and muscle areas using standardized procedures. The cutoff points proposed by the World Health Organization for waist circumference, waist-to-hip ratio, and body mass index were used. Analyses were performed using the Spearman correlation coefficient and linear regression adjusted for age. RESULTS: About 50% of elderly subjects were classified as overweight and more than 50% of women in all age groups had waist circumference and waist-to-hip ratio outside the normal range. Among men, these percentages were about 40% for waist circumference and 20% for waist-to-hip ratio. Among elderly subjects, the partial correlation (adjusted for age) between body mass index and measures of adiposity (waist circumference, skinfolds, and arm fat area) ranged from 0.45 to 0.85 for men and 0.55 to 0.86 for women. Weight and waist circumference were the variables more strongly correlated with body mass index among both elderly persons and adults. CONCLUSIONS: The prevalence of overweight among elderly persons was high for both men and women. Body mass index shows a similar relationship with adiposity regardless of ageing. PMID- 15895134 TI - [Anthropometry of elderly people living in geriatric institutions, Brazil]. AB - OBJECTIVE: To provide anthropometric and body composition information on elderly people living in geriatric institutions. METHODS: Three-hundred and five elderly people, of both sexes, living in six geriatric institutions in Fortaleza were assessed. The following anthropometric variables were studied: weight, height, body mass index, mid-arm circumference, triceps skinfold thickness, arm muscle circumference, and corrected arm-muscle area. The body mass index was calculated as weight divided by the square of the height (m2). The arm muscle circumference and corrected arm-muscle area were calculated using specific equations. The results are presented as means, standard deviations and percentiles (5th, 10th, 25th, 50th, 75th, 90th and 95th). The analyses included Student t-test to detect differences in mean values of the variables between both sexes. Age impact was investigated by ANOVA. RESULTS: In all variables, mean values in men were higher than those in women, except for triceps skinfold thickness . The mean difference of the variables body mass index and mid-arm circumference for both sexes were not statistically significant (p>0.05). Age has significantly contributed to reducing the variables' values. This means that specific reference standards are needed for elderly people. CONCLUSIONS: Despite being institutionalized, there was seen a trend of decreasing anthropometric values in the study population similar to that found in other studies of elderly people but with different values. Thereby, such values could be useful in the nutritional assessment of institutionalized elderly people. PMID- 15895135 TI - [Spatial analysis of socioeconomic determinants of homicide in Brazil]. AB - OBJECTIVE: To investigate the association between homicide rates and socio economic variables taking into account the spatial site of the indicators. METHODS: An ecological study was conducted. The dependent variable was the rate of homicides among the male population aged 15 to 49 years, residing in the districts of the State of Pernambuco from 1995 to 1998. The independent variables were an index of the living conditions, per capita family income, Theil inequality index, Gini index, average income of the head of the family, poverty index, rate of illiteracy, and demographic density. The following techniques were used in the analysis: a spatial autocorrelation test determined by the Moran index, multiple linear regression, a spatial regression model (CAR) and a generalized additive model for the detection of spatial trend (LOESS). RESULTS: The illiteracy and the poverty index explained 24.6% of the total variability of the homicide rates and there was an inverse relationship. Moran's I statistics indicated spatial autocorrelation between municipalities. The multiple linear regression model best fitted for the purposes of this study was the Conditional Auto Regressive (CAR) model. The latter confirmed the association between the poverty index, illiteracy and homicide rates. CONCLUSIONS: The inverse association observed between socio-economic indicators and homicides may be expressing a process that propitiates improvement in living conditions and that is linked predominantly to conditions that generate violence, such as drug traffic. PMID- 15895136 TI - [Blood glucose control in diabetes patients seen in primary health care centers]. AB - OBJECTIVE: To identify factors associated to poor glycemic control among diabetic patients seen at primary health care centers. METHODS: A cross-sectional study was carried out in a sample of 372 diabetic patients attending 32 primary health care centers in southern Brazil. Data on three hierarchical levels of health unit infrastructure, medical care and patient characteristics were collected. RESULTS: The frequency of poor glycemic control was 50.5%. Multivariate analysis (multilevel method) showed that patients with body mass indexes below 27 kg/m2, patients on oral hypoglycemic agents or insulin, and patients diagnosed as diabetic over five years prior to the interview were more likely to present poor glycemic control when compared to their counterparts. CONCLUSIONS: Given the hierarchical data structuring, all associations found suggest that factors associated to hyperglycemia are related to patient-level characteristics. PMID- 15895137 TI - [Social inequality and homicide rates in Sao Paulo City, Brazil]. AB - OBJECTIVE: The relation between income and mortality due to violence has been studied in recent years. The Synthesis of Social Indicators of 2002 [Sintese de Indicadores Sociais, 2002], published by The Brazilian Institute of Geography and Statistics (IBGE), states that one of the most outstanding characteristic of Brazilian society is inequality. The proposal of this ecological study was to test the association between homicide rates, and some health and socioeconomic indicators. METHODS: This is an ecological cross-sectional study. Data regarding Sao Paulo City, Brazil in the year 2000 was analyzed. The association between homicide coefficients and the following five indicators were tested: infant mortality rates, monthly average income of household heads, percentage of adolescents aged 15 to 17 years not attending school, proportion of pregnant adolescent women aged 14 to 17 years and demographic density. Pearson's correlation coefficient and a multiple linear regression model were utilized to test these associations. RESULTS: The municipal homicide rate was 57.3/100,000. The correlation between homicide rates and average monthly income was strong and negative (r=-0.65). Higher homicide rates were found in the districts whose inhabitants had lower incomes and lower rates were found in those districts whose inhabitants had higher incomes. The correlation between homicide rates and proportion of adolescents not attending school was positive and strong (r=0.68). The correlation between homicide rates and the proportion of pregnant adolescent women was positive and strong (r=0.67). The correlation between homicides and the rate of infant mortality was r=0.24 (for all: p<0.05). The correlation between demographic density and homicides was not significant. Although the univariate regression was positive for four indicators, the multivariate regression test was only significant for average monthly income (negative) and proportion of adolescents not attending school (positive) (for both indicators: p<0.01). CONCLUSIONS: The findings highlight the problem of homicides and socioeconomic disparities in S. Paulo City. Economic development and reducing socioeconomic inequality may have an impact on the rates of mortality due to violence. PMID- 15895138 TI - [AIDS mortality and socioeconomic indexes in the city of Sao Paulo, 1994-2002]. AB - OBJECTIVE: To investigate the correlation between AIDS mortality rates and social inclusion/exclusion indexes among male and female populations aged between 25 to 49 years. METHODS: The study was carried out in 96 administrative districts of the municipality of Sao Paulo in the period 1994-2002. Mortality data were collected from the Program for Improving Local Mortality Data and from population estimates based on the 1991 and 2000 census of the State System Data Analysis (SEADE) Institute and the Municipal Department of Planning. The indicators were obtained from the city's map of exclusion (1996 and 2000). Statistical analysis was performed using Pearson's correlation test (at 5% level). RESULTS: A significant positive correlation was seen among men between AIDS mortality and the district life quality index from 1994 to 1998 (p<0.05). Among women, a significant negative correlation (p<0.05) was seen for the whole study period between AIDS mortality and the equity index, which measures the proportion of illiterate women who are family heads. From 2000, it was also observed a significant negative correlation (p<0.05) among women between AIDS mortality and the global social exclusion index. CONCLUSIONS: The study results suggest a shift of AIDS mortality to exclusion areas and might indicate a relationship between AIDS mortality and socioeconomic factors. Further epidemiological and social sciences studies are needed to investigate these associations. PMID- 15895139 TI - [Anthropophilic activity of Aedes aegypti and of Aedes albopictus in area under control and surveillance]. AB - OBJECTIVE: To describe the hematophagous activity of Ae. aegypti and Ae. albopictus in an area under control and surveillance. METHODS: The study was conducted during 18 months, from April, 1993 to October, 1994, in Cosmopolis, Sao Paulo state, Brazil. Human baits were used to collect mosquitoes. The number of females captured is presented monthly by area of the city and local in the household. The rainfall was measured and indices are presented without model adjustment. RESULTS: The presence of females of both species was observed in 83% (Ae albopictus) and 61% (Ae. aegypti) of the period studied. The months of January, February and March presented the highest rates of activity for females of both species, with Ae. albopictus being more frequently captured than Ae. aegypti. Both species were captured in central and peripheral areas of the city, during the day from 9 to 12 am and from 4 to 7 pm. By a Poisson regression, it was observed that Ae. albopictus females were more frequently captured in the peri-housed area, in an independent way considering the area of the city. CONCLUSIONS: It has been identified difference on the hematophagous activity only for Ae. albopictus, being of importance the outside area of the house. PMID- 15895140 TI - [Population statistics of Triatoma rubrovaria in laboratory]. AB - OBJECTIVE: To obtain T. rubrovaria population parameters in order to describe its demographic characteristics. METHODS: The study was carried out in the laboratory of Arthropods, Corrientes, Argentina, from October 2000 to February 2003. Eggs were grouped to form five 100-egg cohorts. Insects were fed on chickens (Gallus domesticus). The cohorts were monitored weekly and kept under controlled temperature (28 +/- 3 degrees C) and relative humidity (63 +/- 10%). A life table was constructed and other vital statistics were calculated and recorded. RESULTS: Higher mortality was recorded in the first through the fourth nymphal stadium. A constant decrease was seen from the fifth nymphal instar. Life expectancy dropped linearly after overcoming the critical stages. Adults mean survival was 50.2 weeks. The first oviposition was after 40.6 weeks. The fecundity was 859.6 eggs with an average 22.8 eggs per female. The reproductive period was 37.7 weeks. The generation time was 55.3 weeks and the net reproduction rate was 133.7. The intrinsic rate of weekly increment was 0.088. In a stable age distribution the population would be composed of 25.3% eggs, 72.3% nymphs and 2.4% adults. Adults accounted for more than 70% of the total reproductive value. CONCLUSIONS: Triatoma rubrovaria had a long survival as imago, a late first reproduction and a low intrinsic rate of natural increase. PMID- 15895141 TI - [Frequency of pathological gambling among substance abusers under treatment]. AB - OBJECTIVE: To investigate the frequency of pathological gambling among alcohol and other substance abusers who sought specialized treatment. METHODS: Seventy four outpatients from three different substance abuse treatment services were interviewed. The South Oaks Gambling Screen (SOGS) scale was used for the diagnosis of pathological gambling. The diagnosis of alcohol and other substances abuse was established according to the DSM-IV criteria and the Short Alcohol Dependence Data (SADD) scale. The Portuguese version of the Self-Report Questionnaire (SRQ) scale was used to detect psychiatric symptoms and the Center for Epidemiological Studies Depression Scale (CES-D) for depressive symptoms. Average scores obtained from the application of these scales were compared using the Student t-test. RESULTS: All subjects met the criteria for drug abuse, 61.6% met the alcohol dependence criteria, 60.3% for cocaine/crack, and 34.2% for cannabis. According to the SOGS scale, the majority of drug addicts (70.3%) were classified as social gamblers, 10.8% as problem gamblers and 18.9% as pathological gamblers. Psychiatric and depression symptoms were found in the sample. Pathological gambling patients showed more depression symptoms than non pathological gambling patients. CONCLUSIONS: A high frequency of pathological gambling was found among the drug addicts interviewed. It is emphasized the importance of investigating pathological gambling among patients under treatment of drug abuse and to include strategies for the treatment of this disorder. PMID- 15895142 TI - [Evaluation of pharmaceutical assistance in public primary care in Brasilia, Brazil]. AB - OBJECTIVE: Pharmaceutical assistance is essential in health care and a right of citizens according to Brazilian law and drug policies. The study purpose was to evaluate aspects of pharmaceutical assistance in public primary health care. METHODS: A cross-sectional study using WHO drug indicators was carried out in Brasilia in 2001. From a random sample of 15 out of 62 centers thirty exiting patients per center were interviewed. RESULTS: Only 18.7% of the patients fully understood the prescription, 56.3% could read it, 61.2% of the prescribed drugs were actually dispensed, and mean duration of pharmaceutical dispensing was 53.2 seconds. Each visit lasted on average 9.4 minutes. Of prescribed and non dispensed drugs, 85.3% and 60.6% were on the local essential drug list (EDL) respectively. On average 83.2% of 40 essential drugs were in stock, and only two centers had a pharmacist in charge of the pharmacy. The mean number of drugs per prescription was 2.3, 85.3% of prescribed drugs were on the EDL, 73.2% were prescribed using the generic denomination, 26.4% included antibiotics and 7.5% were injectables. The most prescribed groups were: cardiovascular drugs (26.8%), anti-infective drugs (13.1%), analgesics (8.9%), anti-asthmatic drugs (5.8%), anti-diabetic drugs (5.3%), psychoactive drugs (3.7%), and combination drugs (2.7%). CONCLUSIONS: Essential drugs were only moderately available almost 30 years after the first Brazilian EDL was formulated. While physician use of essential drugs and generic names was fairly high, efficiency was impaired by the poor quality of pharmaceutical care, resulting in very low patient understanding and insufficient guarantee of supply, particularly for chronic diseases. PMID- 15895143 TI - [Problem-solving approach in the training of healthcare professionals]. AB - OBJECTIVE: To discuss the problem-solving approach in the training of healthcare professionals who would be able to act both in academic life and in educational practices in services and communities. METHODS: This is an analytical description of an experience of problem-based learning in specialization-level training that was developed within a university-level healthcare education institution. The analysis focuses on three perspectives: course design, student-centered learning and the teacher's role. RESULT: The problem-solving approach provided impetus to the learning experience for these postgraduate students. There was increased motivation, leadership development and teamworking. This was translated through their written work, seminars and portfolio preparation. The evaluation process for these experiences presupposes well-founded practices that express the views of the subjects involved: self-assessment and observer assessment. The impact of this methodology on teaching practices is that there is a need for greater knowledge of the educational theories behind the principles of significant learning, teachers as intermediaries and research as an educational axiom. CONCLUSIONS: The problem-solving approach is an innovative response to the challenges of training healthcare professionals. Its potential is recognized, while it is noted that educational innovations are characterized by causing ruptures in consolidated methods and by establishing different ways of responding to demands presented at specific moments. The critical problems were identified, while highlighting the risk of considering this approach to be a technical tool that is unconnected with the design of the teaching policy. Experiences and analyses based on the problem-solving assumptions need to be shared, thus enabling the production of knowledge that strengthens the transformation of educational practices within healthcare. PMID- 15895144 TI - [Prevalence of noise-induced hearing loss in metallurgical company]. AB - OBJECTIVE: To analyze the prevalence of cases suggestive of noise-induced hearing loss (NIHL) among metalworkers who were potentially exposed to occupational noise (from 83 to 102 dB). METHODS: A cross-sectional study was carried out in a metalworking company providing services in Rio de Janeiro, Brazil. Clinical and occupational data on 182 workers who were active between November 2001 and March 2002 were obtained from the company's Hearing Conservation Program and analyzed. In order to characterize the noise exposure status within the work environment, the acoustic classifications from the operating units of the client companies were used, due to difficulty in quantification at an individual level. Associations between these cases and variables such as age, length of service in the company, length of exposure to occupational noise and degree of use of individual protection equipment were tested by means of prevalence ratios and logistic regression analysis. RESULTS: The prevalence of cases suggestive of NIHL was 15.9% and significant associations (p<0.05) were identified from multivariate analysis between these cases and the variables of age and degree of use of individual protection equipment. CONCLUSIONS: The results found contribute towards better understanding of the behavior of some of the main characteristics of NIHL, in a particular situation of the organizing of work that is relatively common in the Brazilian industrial context. PMID- 15895145 TI - [Job control, job demands, social support at work and health among adolescent workers]. AB - OBJECTIVE: To evaluate physical and psychological dimensions of adolescent labor (such as job demands, job control, and social support in the work environment), and their relation to reported body pain, work injuries, sleep duration and daily working hours. METHODS: A total of 354 adolescents attending evening classes at a public school in Sao Paulo, Brazil, answered questionnaires regarding their living and working conditions (Karasek's Job Content Questionnaire, 1998), and their health status. Data collection took place in April and May 2001. Multiple logistic regression analysis was used to determine relations among variables. RESULTS: Psychological job demands were related to body pain (OR=3.3), higher risk of work injuries (OR=3.0) and reduced sleep duration in weekdays (Monday to Thursday) (p<0.01). Lower decision authority in the workplace (p=0.03) and higher job security (p=0.02) were related to longer daily working hours. CONCLUSIONS: It was concluded that besides physical stressors, psychological factors are to be taken into account when studying adolescent working conditions, as they may be associated with negative job conditions and health effects. PMID- 15895146 TI - [Underreporting of non-fatal work-related injuries in Brazil]. AB - OBJECTIVE: The implementation of work-related injury prevention policies has been hindered by underreporting of incidents among formal workers, and substantial underreporting among informal workforce. This study aimed at estimating the underreporting of work-related injury in a median-sized city. METHODS: A random survey was carried out among residences in the urban area of Brazil. Residents were interviewed about the occurrence of work-related injury to people aged more than nine years in the last 90 days. All incidents reported were double checked in the National Social Security Institute (INSS) records. RESULTS: There were 9,626 residences visited. It was estimated 79.5% (CI 95%: 78.8%-80.3%) of underreporting of work-related injury. CONCLUSIONS: Work-related injury reporting is poor in the study location and this may be occurring in other cities. Data suggest the need to build up information systems on Brazilian workers' health. It should incorporate methods, materials and human recourses necessary to recognize, store, analyze, and spread information to support injury prevention policies and promote workers' health programs. PMID- 15895147 TI - [The meanings attributed to the voice by menopause women]. AB - OBJECTIVE: To analyze the meanings attributed to the voice by menopausal women. METHODS: Data were collected from 148 medical records of women seen in the Climacteric Women Health Program of the Irmandade Santa Casa de Misericordia Gynecology Outpatient Clinic in Curitiba, Brazil, between February 2000 and October 2001. Of them, 30 women aged between 48 and 59 years who did not consider themselves as voice professionals and had been at least 12 months in amenorrhea were selected. They were interviewed using a thematic guide with semi-structured questions. Interviews were recorded and then transcribed. Their description, analysis, and interpretation were based on social representation of the collective individual's discourse through a hermeneutics-dialectics approach. RESULTS: The resulting 27 collective discourses showed that women related their voice to biological and psychological characteristics and social aspects of daily life, identifying vocal changes during their life course. CONCLUSIONS: Social representations of communicational and functional nature were observed and highlight voice as an element of personal identity construction conceived in the social sphere. There is a need for further research based on social sciences as well as epidemiological studies and to examine voice therapy in a frailer larynx and also prioritize voice health as part of comprehensive health care delivered to climacteric women. PMID- 15895148 TI - [Knowledge, attitudes and practice related to Papanicolaou smear test among Argentina's women]. AB - OBJECTIVES: To evaluate the knowledge, attitudes, and practice regarding the Papanicolaou test in an Argentinean community. METHODS: Two hundred women were interviewed at their homes in Puerto Leoni, Misiones, Argentina. Women were selected by simple random sampling. Answers were described in terms of knowledge, attitudes, and practice, ant their respective adequacies with respect to the Papanicolaou test, as previously defined. Adequacy was compared between the categories of the control variables by chi2 test with a 5% significance level. RESULTS: Knowledge and practice of the Papanicolaou test were adequate in 49.5% and 30.5% of subjects, respectively, although the attitude towards the test was considered adequate in 80.5% of subjects. Another important finding was that women reported as the main reason for not undergoing the test the lack of a request by a physician or healthcare professional. CONCLUSIONS: Our results show a need for increasing the information provided to the public, especially by healthcare services and professionals, thus generating knowledge among the population of the advantages and benefits of Papanicolaou testing. PMID- 15895149 TI - [Impact of health services delivery on hospital admission utilization in Brazil]. AB - OBJECTIVE: To assess hospital services utilization in Brazil incorporating information about health services delivery. METHODS: Data from the National Household Survey carried out by Brazilian Institute of Statistics and Geography (IBGE) in 1998 and from other sources were collected. Hierarchical models having the individual in the first level and the state of residence in the second level were used. Two models were separately adjusted for adults and children: logistic regression was used when to have been or not admitted was the response variable, and Poisson's regression was used when the number of admissions was the response variable. This last model was adjusted only for those individuals who had been admitted at least once. RESULTS: The main determinant of hospital admissions was health need. Poor people were more likely to be admitted when controlling for health needs and enabling factors (health insurance coverage and regular health service). Only 1 to 3% of the variability in hospital admission utilization could be attributed to differences in services delivery at the state level. In the logistic models, the number of beds was positively associated and the number of doctors was negatively associated with the likelihood of admission. Poisson's models did not show any delivery variables associated with the likelihood of admission. CONCLUSIONS: These results suggest a delivery-induced demand impact concerning hospital beds. The inverse association of the number of doctors and hospital admissions suggests the impact of outpatient care on hospital utilization. PMID- 15895150 TI - [Evaluation of simplified DMFT indices in epidemiological surveys of dental caries]. AB - OBJECTIVE: To analyze whether two simplified indices, "DMF in 6 Teeth"(DMF6T) and "DMF in two quadrants" (DMF2Q), could be indicated for epidemiological surveys of oral health according to dental caries distribution. METHODS: The sample came from epidemiological data in 29 municipalities. A total of 2,378 examinations were done on 12-year-old schoolchildren. The mean DMFT from each locality was utilized to obtain three DMFT prevalence groups (low, moderate and high), for which the simplified indices were calculated. Statistical analysis was done using intraclass correlation and the Wilcoxon and chi-squared tests, with a significance level of 5%. RESULTS: Intraclass correlation between the DMFT index and the simplified indices ranged from 0.82 to 0.95 (p<0.05). No significant differences in the prevalence of the studied parameters were observed between the means of the DMFT and DMF2Q indices (p>0.05), although DMF6T showed differences. The proportions of decayed, missed and filled teeth were similar between the DMFT and DMF2Q indices (p>0.05). CONCLUSIONS: The simplified DMF2Q index can be used in epidemiological surveys in areas with low, moderate and high prevalence of dental caries. However, better evaluation of the DMF6T index needs to be made. PMID- 15895151 TI - [Toxocara sp. eggs and Ancylostoma sp. larva in public parks, Brazil]. AB - Visceral and cutaneous larva migrans are parasitic zoonoses caused by the infection of larval nematodes Toxocara sp. and Ancylostoma sp. respectively. The objective of this study was to investigate the contamination by Toxocara sp. eggs and Ancylostoma sp. eggs and larva of soil samples collected from public parks and children's playground areas in state of Minas Gerais, Brazil, using both Baermann's method and centrifugal flotation technique. Toxocara sp. and Ancylostoma sp. eggs were observed in soil samples collected from public squares in 17.4% (4/23) and 69.6 (16/23) respectively. In schools and child day care settings the contamination by Ancylostoma sp. larva in sand samples was 11.1% (2/18). Public parks are settings of more potential risk of Toxocara sp. eggs and Ancylostoma sp. infection. Stool parasitology testing of 174 stool samples showed 58% and 23% of Ancylostoma sp and Toxocara sp eggs infection respectively. PMID- 15895152 TI - [Impact of interventions promoting condom use among HIV-infected individuals]. AB - In order to identify educational interventions promoting male condom use during intercourse among HIV+ persons, we conducted a systematic review of publications. Lilacs/Bireme, Medline and Popline data sets and CDC and UNAIDS sites were searched without time limit. Uniterms used were: women; men; interventions; HIV; Aids; HIV positive; risk behaviors; sexual risk behaviors; and intervention studies. Fourteen interventions were included, eight of which reported a positive result. Most frequent methodological weaknesses observed were lack of randomization, no control for confounding, high rates of losses to follow-up, small sample sizes, and outcome of condom use stated by patient self-report. Publication bias favoring studies showing a beneficial effect has to be considered. Effective interventions aiming to promote condom use among HIV positive persons are currently an important tool in the prevention of HIV dissemination. PMID- 15895153 TI - [Outside clinical setting experience in a public hospital and oral health promotion]. AB - OBJECTIVE: To describe a teaching aid activity aimed at providing experience for promoting collective oral health to graduating dental students. METHODS: This experience was based on the evaluation of students' performance as oral health educators as they had, among other duties, to motivate inpatients and their families to practice healthy habits aiming at a comprehensive and more human care of hospitalized patients. RESULTS: The results show that oral hygiene was incorporated into hospital routine, evidenced by the differences between baseline and final dental biofilm indices (1.72 and 1.17, respectively). Using the U-Mann Whitney test, this difference was extremely significant (p<0.001) and reveals that mother-child were highly motivated with respect to oral hygiene. CONCLUSIONS: It is concluded that teaching-learning experiences derived from interdisciplinary and interdisciplinary activities have allowed for a better understanding of the health-disease process by dental students. It is also an opportunity for learning about planning and implementing education-prevention activities, which complement these students' technical-professional experience and promotes social sensitivity, which is essential to professional training. PMID- 15895154 TI - [Geriatric outpatient healthcare: hierarchical demand structuring]. AB - In Brazil, the rapid growth of the elderly population has been causing a great impact on the healthcare system, with increased costs and service utilization. The inefficiency of traditional models for geriatric healthcare has made it essential to change the healthcare concepts for this population. This can take place through the development of new healthcare models that include the means to identify, assess and treat elderly patients with a variety of morbid and functional conditions, and which can be applied diverse healthcare scenarios. An outpatient model is proposed, with two stages that differ in the depth and coverage of their actions. These stages are organized as increasing levels of complexity and are capable of selecting subgroups of individuals that, because of their risk characteristics, should follow different paths through the healthcare structure. This paper discusses the first stage of this model, which involves risk identification among large groups of elderly people, by means of structuring a hierarchical flow of actions and using assessment tools of adequate sensitivity and specificity. Individuals aged 65 years or over who are detected through walk in outpatient consultation, home visits or telephone interview are classified using a rapid screening risk evaluation instrument composed of eight items. Depending upon the level of risk presented, the individual will either be referred to another level of functional evaluation (medium-high and high risk levels), or to normal clinical care and old people's community centers (low and medium risk levels). The second stage will be the subject of a subsequent paper. PMID- 15895155 TI - [Use of standardized questionnaire in epidemiological survey of asthma]. PMID- 15895156 TI - [Investigation of an outbreak of typhoid fever in Santos, SP, Brazil 2004]. PMID- 15895160 TI - Golden mussel Limnoperna fortunei (Bivalvia: Mytilidae) distribution in the main hydrographical basins of Uruguay: update and predictions. AB - Limnoperna fortunei, an Asiatic rivers bivalve has become a worldwide problematic invasive species causing several water quality and macrofouling problems. In the Neotropical region it was first recorded in 1991 in the Rio de la Plata coast, Buenos Aires province. Since this, it showed a quick upstream invasion into the principals aquatic systems of the Plata Basin. Nevertheless, there is not a study about its invasion and distribution process in aquatic systems of Uruguay. We describe the new records of Limnoperna fortunei in Uruguayan coast of Rio de la Plata, Santa Lucia, Negro and Uruguay Rivers. With these results we aim to estimate its distributional limits for Uruguay main hydrographical basins. We also deal with the role of salinity as the main abiotic factor in limiting the east distribution of this mussel in Uruguayan coast of Rio de la Plata and as a potential determinant of the "new" colonization on the Atlantic and the Merin Lagoon Basins. Its presence in the ecosystems not only can cause changes at the ecosystem level but also endanger the associated community, favoring the displacement and the disappearance of endemic species. PMID- 15895161 TI - Evidence for a role of nitric oxide in hindlimb vasodilation induced by hypothalamic stimulation in anesthetized rats. AB - Electrical stimulation of the hypothalamus produces cardiovascular adjustments consisting of hypertension, tachycardia, visceral vasoconstriction and hindlimb vasodilation. Previous studies have demonstrated that hindlimb vasodilation is due a reduction of sympathetic vasoconstrictor tone and to activation of beta2 adrenergic receptors by catecholamine release. However, the existence of a yet unidentified vasodilator mechanism has also been proposed. Recent studies have suggested that nitric oxide (NO) may be involved. The aim of the present study was to investigate the role of NO in the hindquarter vasodilation in response to hypothalamic stimulation. In pentobarbital-anesthetized rats hypothalamic stimulation (100 Hz, 150 microA, 6 s) produced hypertension, tachycardia, hindquarter vasodilation and mesenteric vasoconstriction. Alpha-adrenoceptor blockade with phentolamine (1.5 mg/kg, iv) plus bilateral adrenalectomy did not modify hypertension, tachycardia or mesenteric vasoconstriction induced by hypothalamic stimulation. Hindquarter vasodilation was strongly reduced but not abolished. The remaining vasodilation was completely abolished after iv injection of the NOS inhibitor L-NAME (20 mg/kg, iv). To properly evaluate the role of the mechanism of NO in hindquarter vasodilation, in a second group of animals L-NAME was administered before alpha-adrenoceptor blockade plus adrenalectomy. L-NAME treatment strongly reduced hindquarter vasodilation in magnitude and duration. These results suggest that NO is involved in the hindquarter vasodilation produced by hypothalamic stimulation. PMID- 15895162 TI - Meristematic activity of the Endodermis and the Pericycle in the primary thickening in monocotyledons: considerations on the "PTM". AB - This paper proposes a new interpretation for primary thickening in monocotyledons. The anatomy of the vegetative organs of the following species was examined: Cephalostemon riedelianus (Rapataceae), Cyperus papyrus (Cyperaceae), Lagenocarpus rigidus, L. junciformis (Cyperaceae), Echinodorus paniculatus (Alismataceae) and Zingiber officinale (Zingiberaceae). The endodermis with meristematic activity was observed in the root of all the species, in the stem of Cyperus, Cephalostemum and Lagenocarpus rigidus, and in the leaf trace of Cyperus and leaf of Echinodorus. Considering the continuity of tissues through the root, stem and leaf, the authors conclude that in the stem the pericycle remains active throughout the life of the plant as the generator of the vascular tissue. The "Primary Thickening Meristem" is in fact the pericycle plus the endodermis and its derivatives (or only the pericycle). Close to the stem apex, the assemblage of seems to be a unique meristem, giving rise to the inner cortex and vascular tissues. PMID- 15895163 TI - Targeting exosites on blood coagulation proteases. AB - The high specificity of blood coagulation proteases has been attributed not only to residues surrounding the active site but also to other surface domains that are involved in recognizing and interacting with macromolecular substrates and inhibitors. Specific blood coagulation inhibitors obtained from exogenous sources such as blood sucking salivary glands and snake venoms have been identified. Some of these inhibitors interact with exosites on coagulation enzymes. Two examples are discussed in this short revision. Bothrojaracin is a snake venom-derived protein that binds to thrombin exosites 1 and 2. Complex formation impairs several exosite-dependent activities of thrombin including fibrinogen cleavage and platelet activation. Bothrojaracin also interacts with proexosite 1 on prothrombin thus decreasing the zymogen activation by the prothrombinase complex (FXa/FVa). Ixolaris is a two Kunitz tick salivary gland inhibitor, that is homologous to tissue factor pathway inhibitor. Recently it was demonstrated that ixolaris binds to heparin-binding exosite of FXa, thus impairing the recognition of prothrombin by the enzyme. In addition, ixolaris interacts with FX possibly through the heparin-binding proexosite. Differently from FX, the ixolaris-FX complex is not recognized as substrate by the intrinsic tenase complex (FIXa/FVIIIa). We conclude that these inhibitors may serve as tools for the study of coagulation exosites as well as prototypes for new anticoagulant drugs. PMID- 15895164 TI - Ouabain exacerbates activation-induced cell death in human peripheral blood lymphocytes. AB - Lymphocytes activated by mitogenic lectins display changes in transmembrane potential, an elevation in the cytoplasmic Ca2+ concentrations, proliferation and/or activation induced cell death. Low concentrations of ouabain (an inhibitor of Na+,K+-ATPase) suppress mitogen-induced proliferation and increases cell death. To understand the mechanisms involved, a number of parameters were analyzed using fluorescent probes and flow cytometry. The addition of 100 nM ouabain to cultures of peripheral blood lymphocytes activated with 5 microg/ml phytohemagglutinin (PHA) did not modify the increased expression of the Fas receptor or its ligand FasL induced by the mitogen. However, treatment with ouabain potentiated apoptosis induced by an anti-Fas agonist antibody. A synergy between ouabain and PHA was also observed with regard to plasma membrane depolarization. PHA per se did not induce dissipation of mitochondrial membrane potential but when cells were also exposed to ouabain a marked depolarization could be observed, and this was a late event. It is possible that the inhibitory effect of ouabain on activated peripheral blood lymphocytes involves the potentiation of some of the steps of the apoptotic process and reflects an exacerbation of the mechanism of activation-induced cell death. PMID- 15895165 TI - Vaccines based on the cell surface carbohydrates of pathogenic bacteria. AB - Glycoconjugate vaccines, in which a cell surface carbohydrate from a micro organism is covalently attached to an appropriate carrier protein are proving to be the most effective means to generate protective immune responses to prevent a wide range of diseases. The technology appears to be generic and applicable to a wide range of pathogens, as long as antibodies against surface carbohydrates help protect against infection. Three such vaccines, against Haemophilus influenzae type b, Neisseria meningitidis Group C and seven serotypes of Streptococcus pneumoniae, have already been licensed and many others are in development. This article discusses the rationale for the development and use of glycoconjugate vaccines, the mechanisms by which they elicit T cell-dependent immune responses and the implications of this for vaccine development, the role of physicochemical methods in the characterisation and quality control of these vaccines, and the novel products which are under development. PMID- 15895170 TI - Retrospective study of 151 patients with cutaneous leishmaniasis treated with meglumine antimoniate. AB - We retrospectively analyzed a series of 151 cases of cutaneous leishmaniasis treated between 1967 and 1982. One-hundred-and-thirty-nine (92%) patients presented with active lesions and were treated with daily doses of meglumine antimoniate: 81 adults received a 5-ml vial IM and 58 children received 1 to 5 ml. Forty-five (32.4%) patients underwent continuous treatment with meglumine antimoniate for 25 to 116 days without rest intervals, and 94 (67.6%) intermittent treatment with 2 to 5 series of meglumine antimoniate. Intermittent series could include schedules of daily IM applications for 10 to 25 days each and intervals varying from 10 to 60 days. Antimony dose was calculated for 66 (47.5%) patients and ranged from 3.9 to 28.7 Sb5+/kg/day. Of these, 35 patients received > or =10 mg and 31 patients < 10 mg Sb5+/kg/day. Median time of healing was longer for lesions on the legs and feet -- 67.5 days versus 48.7 days (p < 0.001) for other sites. However, there were no significant differences in the median time of healing between adults and children, intermittent and continuous regimens or high and low antimony doses. Fifty-one patients were reassessed 5 to 14 years after treatment and showed no evidence of disease. These results support further investigation (clinical trials) on treatment using low doses of antimony. PMID- 15895171 TI - [Seroprevalence of hepatitis B and malaria infection in Labrea, Brazilian western Amazon: estimates of coinfection rates]. AB - The Amazon region is known for the high occurrence of hepatitis B virus (HBV) infection, and accounts for more than 98% of malaria cases in Brazil. Despite the controversy, it has been proposed that when associated they may lead to important effects in the natural history of both infections. This study estimates the prevalence of coinfection within general population of an endemic region of HBV and malaria in the Brazilian Amazon. The prevalence of HBsAg was 3.3% (95% CI 2.1%-5.1%,) and total anti-HBc 49.9% (95% CI 45.9%-53.8%). The prevalence of antibodies against Plasmodium vivax and Plasmodium falciparum antigens was 51.4% (311/605) (95% CI 47.3%-55.4%). Related to the simultaneous presence of malaria antibodies and HBV serological markers, in 1.8% (11/605), (95% CI 1.0%-3.3%), the presence of HBsAg was also demonstrated, mean age 26 years (p <0.001). This study points to similarities in the distribution of these diseases, such as the occurrence mainly among young adults. The events may occur in different times. Also shown differences such as the low risk of malaria in the group up to fifteen years, where HBV circulates with moderate intensity. The prevalence of HBV and malaria coinfection is in fact less than the rates of HBsAg carriage, showing a heterogeneous pattern related to the clinical spectrum of HBV infection. PMID- 15895172 TI - Bancroftian filariasis in an endemic area of Brazil: differences between genders during puberty. AB - Gender differences in susceptibility to infectious diseases have been observed in various studies. A survey was performed in a bancroftian filariasis endemic area in the city of Olinda, Brazil. All residents aged 5 years or older were examined by thick blood film. People aged 9 to 16 years were interviewed and also tested for filarial antigenaemia. Data were analyzed by contingency table methods and regression models. The risk of microfilaraemia for males was significantly higher. Among those aged 9 to 16 years, the analysis of gender and filariasis by age showed that boys from 15 to 16 years had a higher risk of infection than girls. No association was found between menarche and filariasis in girls. The data suggest that variations between gender in filariasis could result, at least in part, from an increase in susceptibility of men. This epidemiologic feature needs to be considered while formulating elimination plans. PMID- 15895173 TI - [American tegumentary leishmaniasis caused by Leishmania (Viannia) braziliensis in military training area of Zona da Mata in Pernambuco]. AB - The aim of work was to study the epidemiology of American tegumentary leishmaniasis in a military training unit situated in Zona da Mata region of Pernambuco State. Between 2002 and 2003 twenty-three cases were notified by clinical exam, detection and/or isolation of parasite and Montenegro skin test. Seven stocks of Leishmania (Viannia) braziliensis were obtained from patients, identified by a panel of specific monoclonal antibodies and isoenzymatic electrophoresis profiles. An epidemiologic survey on prevalence of infection was carried out by Montenegro skin test in the population that underwent training activities during the same period, out of which 25.3% were identified as positive. These results in association with previous data from this area, shows the maintenance of a primary transmission cycle and the occurrence of periodical outbreaks after training activities in local areas of remnant Atlantic rain forest. PMID- 15895174 TI - [Clinical-epidemiologic profile of paracoccidioidomycosis at the Stomatology Department of Sao Lucas Hospital, Pontificia Universidade Catolica of Rio Grande do Sul]. AB - The aim of the article was to describe clinical and epidemiological features of 61 oral paracoccidioidomycosis cases from the Stomatology Department of Sao Lucas Hospital, analyzed from July 1976 to June 2004. The State of Rio Grande do Sul is an endemic region due to ecoepidemiologic conditions, that are favorable for the development of the yeast. Men are the most affected gender, with 58 (95%) affected compared to 3 (5%) women. Ii is most prevalent (70.5%) between 40 and 59 years of age. Agricultural labor was the most common occupation with 27/61 (44.3%) patients. Tobacco smoking was prevalent in 52 (85.3%) of the sample, and alcoholism was a frequent finding. All the patients showed stomatologic manifestations, with ulcers and mulberry-like stomatitis the most prevalent, these oral lesions were also observed in many anatomical sites. Alterations were present in 32 (65.3%) of the pulmonary radiographs. PMID- 15895175 TI - [Aedes (Stegomyia) aegypti (Linnaeus, 1762) breeding sites in native bromeliads in Vitoria City, ES]. AB - Some insects that are vectors of human diseases have accompanied man in his migrations throughout the world and breed exclusively in the proximity of human dwellings. The mosquito Aedes aegypti has been responsible for epidemics of dengue in Brazil and its presence also constitutes a serious risk for future outbreaks of urban yellow fever. The failure of campaigns to eradicate this species justifies the search for alternative breeding sites, which may be beyond the reach of present control measures. In this study the occurrence of Aedes aegypti breeding sites in native bromeliads on rocky slopes was investigated in five areas of Vitoria, capital of the Brazilian State of Espirito Santo, ES. Water contained in the bromeliads was collected with the aid of a suction apparatus to search for culicid larvae. The degree of infestation of buildings in adjacent urban areas was evaluated simultaneously. Culicid larvae were found in bromeliads in four of the five areas investigated, Aedes aegypti being present in two areas. The presence of breeding sites in bromeliads was not related to indices of infestation of buildings in adjacent areas. Further studies are necessary to define whether breeding sites in bromeliads constitute primary foci of Aedes aegypti, or are a consequence of high infestation levels in urban areas. PMID- 15895176 TI - The role of integrated home-based care in patient adherence to antiretroviral therapy. AB - Non-adherence is one of the primary obstacles to successful antiretroviral therapy in HIV+ patients worldwide. In Brazil, the Domiciliary Therapeutic Assistance is a multidisciplinary and integrated home-based assistance program provided for HIV+ patients confined in their homes due to physical deficiency. This study investigated ADT's ability to monitor and promote appropriate adherence to ARV therapy. Fifty-six individuals were recruited from three study groups: Group 1 -- patients currently in the ADT program, Group 2 -- 21 patients previously treated by the ADT program, and Group 3 -- 20 patients who have always been treated using conventional ambulatory care. Using multivariable self reporting to evaluate adherence, patients in the ADT program had significantly better adherence than patients in ambulatory care (F = 6.66, p = 0.003). This effect was independent of demographic and socioeconomic characteristics as well as medical history. Patients in the ADT program also showed a trend towards greater therapeutic success than ambulatory patients. These results suggest the incorporation of characteristics of ADT in conventional ambulatory care as a strategy to increase adherence to ARV therapy. PMID- 15895177 TI - [Identification of places with potential transmission of dengue fever in Porto Alegre using Geographical Information Systems]. AB - The social-environmental conditions that promote the continuity of Aedes aegypti in urban areas and its capacity for dengue transmission have yet to be determined. The aim of this work was to locate dengue cases and vector presence, then to identify social-environmental factors that characterize these sites, through Geographical Information Systems (GIS), with a view to developing a dengue fever prevention model. The vector was mainly located in south and east areas of the city, with a broad dispersion, while most of the cases were located in the central region of the city. The sectors with cases present characteristics of high income. Otherwise, sectors with vector present a predominance of houses and good sanitation infrastructure. The difference between cases and vector spatial patterns ensured for 2002 an absence of dengue transmission in the city. PMID- 15895178 TI - [Triatoma rubrovaria (Blanchard 1843): life table of nymphs, duration of the forms and posture of the females]. AB - The authors followed the development of 150 eggs of Triatoma rubrovaria to enable the construction of a life table for the nymphs. The winged form was reached by 94.64% of the individuals, of which 79 were males and 65 females. The mean lifetime of the males was 115 days and 99 days for the females. Part of this group was divided into 30 couples and kept separated to facilitate the count of egg laying by each female. Another lot was also formed of 30 males and 30 females but these were housed together in a glass crystallizer that allowed the observation of a larger number of eggs laid (7,832 eggs) in relationship to that of the females with only one male (total 5,167 eggs). The good development of the individuals and reproduction of the species in the experimental conditions demonstrated the capacity to maintain colonies of Triatoma rubrovaria in laboratory conditions. PMID- 15895179 TI - [Purpura in patient with disseminated strongiloidiasis]. AB - The association of systemic corticosteroid therapy and dissemination of Strongyloides stercoralis has been increasingly documented in the literature. Skin involvement in disseminated strongyloidiasis has been reported and the most commonly described cutaneous manifestations are rash and petechial eruptions. We present a case of an immunosuppressed man that developed disseminated strongyloidiasis with extensive purpura. PMID- 15895180 TI - [A fatal case of mucocutaneous leishmaniasis after pentavalent antimonial use]. AB - The authors report a case of mucocutaneous leishmaniasis in a 45-year-old patient who was unsuccessfully treated with pentavalent antimonial for 30 days. After 10 days from the initial treatment and before starting a new therapeutic regimen with the same drug the patient died due to sudden cardiac arrest. PMID- 15895181 TI - [Evaluation of the API20C AUX system for the identification of clinically important yeasts]. AB - Fifty yeasts belonging to the genera Candida, Pichia, Saccharomyces, Cryptococcus, Rhodotorula and Trichosporon were identified by classical methods and by the API 20C AUX system. The commercial system correctly identified 92% of the species, with the need for additional tests in 16% of cases. The results were interpreted with good, very good and excellent identification. PMID- 15895182 TI - [Phlebotomine sandflies in fragments of rain forest in Recife, Pernambuco State]. AB - An investigation was conducted into the distribution of sandfly fauna in 4 fragments of Atlantic forest in the Metropolitan Area of Recife. It consisted of the capture adult insects using CDC light traps. A total of 1,173 specimens were distributed in 11 species of Lutzomyia: Lutzomyia evandroi, Lutzomyia choti, Lutzomyia walkeri, Lutzomyia umbratilis, Lutzomyia brasiliensis, Lutzomyia sordellii, Lutzomyia claustrei, Lutzomyia wellcomei, Lutzomyia fluviatilis, Lutzomyia furcata e Lutzomyia aragaoi. PMID- 15895183 TI - [Children day care center: exposition or protection environment to intestinal parasites infestation in Aracaju, SE]. AB - The work aimed to evaluate whether child daycare centers are an environment that protects against or exposes children to intestinal parasite infestation. Stool samples were analyzed from children attending such centers and a control group. It was concluded that attending daycare centers is related to intestinal parasitosis (630% vs. 41.4 %; p <0.01) and the risk of infestation is 1.5 times higher. PMID- 15895184 TI - [Adaptation of the Rugai et al. method for analysis of soil parasites]. AB - This study offers an adaptation of the Rugai method for the collection of cysts, eggs and parasite larvae from beaches, parks and public squares. PMID- 15895185 TI - [Epidemiological aspects of a malaria focus in the districts of Sao Luis, MA]. AB - The study was developed in the neighborhood of Residencial Paraiso, Sao Luis, Maranhao, Brazil, from 1999 to 2001. The aim was to analyze the epidemiological aspects of malaria. The location is receptive and vulnerable to malaria. There were 129 cases. Men from 20 to 29 years of age were most involved. Plasmodium vivax was the agent of all cases and Anopheles aquasalis was the vector. There was no evidence of transmission occurring outside of the houses. PMID- 15895186 TI - [Infectology dimension]. PMID- 15895187 TI - [Melioidosis in a little rural community of Ceara State]. PMID- 15895188 TI - [Myeloradiculopathy in acute schistosomiasis mansoni]. PMID- 15895189 TI - [Gergorio Volcan]. PMID- 15895190 TI - The Women's Health Initiative conundrum. AB - The WHI has been designed to evaluate the metabolic risks and benefits of Estrogen/Progestagen Therapy (HT) or Estrogen Therapy (ET) in women in their later postmenopause. It has not been designed to study the effect of HT or ET on symptomatic peri- and early postmenopausal women. Furthermore, the selection criteria used in the WHI are not congruent with the profiles of women treated in daily medicine by HT/ET: women starting HT/ET in clinical routine are younger, less obese and healthier than the WHI population. Therefore, the results and the risk-benefit-conclusions of the WHI cannot be applied to normal symptomatic peri- and immediately postmenopausal women, and even less to women with early (40-50 years) or premature (40 yrs.) menopause. PMID- 15895191 TI - The design and implementation of the immune epitope database and analysis resource. AB - Epitopes are defined as parts of antigens interacting with receptors of the immune system. Knowledge about their intrinsic structure and how they affect the immune response is required to continue development of techniques that detect, monitor, and fight diseases. Their scientific importance is reflected in the vast amount of epitope-related information gathered, ranging from interactions between epitopes and major histocompatibility complex molecules determined by X-ray crystallography to clinical studies analyzing correlates of protection for epitope based vaccines. Our goal is to provide a central resource capable of capturing this information, allowing users to access and connect realms of knowledge that are currently separated and difficult to access. Here, we portray a new initiative, "The Immune Epitope Database and Analysis Resource." We describe how we plan to capture, structure, and store this information, what query interfaces we will make available to the public, and what additional predictive and analytical tools we will provide. PMID- 15895192 TI - Diagnostic value of measuring liver volume for detecting occult hepatic metastases from colorectal or gastric cancer. AB - Some studies have shown reduced portal blood flow in patients with occult hepatic metastases, which may lead to decreased liver volume. A retrospective study was conducted in patients undergoing curative resection for colorectal (n = 63) or gastric (n = 52) cancer. The ratio of the preoperative computed tomography (CT) estimated liver volume to the standard liver volume (CV/SV ratio) was calculated. The mean +/- SD CT-estimated liver volume was 858 +/-109 in 14 patients who subsequently developed hepatic metastases and 1173 +/- 230 ml in 101 patients without metastases (p < 0.0001). The CV/SV ratio was smaller in patients with metachronous hepatic metastases than in those without (0.78 +/- 0.08 vs. 1.02 + 0.13; p < 0.0001). The results suggest that the liver with occult metastases decreases in size before metastases develop that are detectable using conventional imaging techniques. The CV/SV ratio may be of value in detecting occult hepatic metastases from colorectal and gastric cancer. PMID- 15895193 TI - Clinical significance of occult metastatic melanoma in sentinel lymph nodes and other high-risk factors based on long-term follow-up. AB - Selective sentinel lymphadenectomy (SSL) following preoperative lymphoscintigraphy is the most significant recent advance in the management of patients with primary melanoma. This study evaluates the prognostic value of sentinel lymph node (SLN) status and other risk factors in predicting survival and recurrence in patients with primary cutaneous melanoma. From October 1993 to July 1998 a series of 412 patients with primary invasive melanoma underwent SSL at the UCSF/ Mt. Zion Melanoma Center. The outcome of 363 evaluable patients is summarized in this study. The factors related to survival and disease recurrence were analyzed by Cox proportional hazard regression models. The overall incidence of patients with positive SLNs was 18%. Over a median follow-up of 4.8 years, the overall mortality rate in patients with primary cutaneous melanoma was 18.7%, and 74 recurrences occurred (20.4%). Mortality was significantly related to SLN status [HR = 2.06; 95% Confidence interval (CI) 1.18, 3.58], angiolymphatic invasion (HR = 2.21; 95% CI 1.08, 4.55), ulceration (HR = 1.79; 95% CI 1.02, 3.15), mitotic index (HR =1.38; 95% CI 1.01, 1.90), and tumor thickness (HR = 2.20, 95% CI 1.21, 3.99). Factors significantly related to disease-free survival included SLN status (HR = 2.09; 95% CI 1.31, 3.34), tumor thickness (HR = 1.89; 95%. CI 1.20,2.98), and age (HR= 1.26 95% CI 1.08, 1.47). SLN status was the most significant factor for melanoma recurrence and death. Other important predictors include tumor thickness, ulceration, lymphatic invasion, and mitotic index. PMID- 15895194 TI - Safe and rapid laparoscopic access--a new approach. AB - Despite numerous recent technical advances in minimally invasive surgical technique, the potential exists for serious morbidity during initial laparoscopic access. Safe access depends on adhering to well-recognized principles of trocar insertion, knowledge of abdominal anatomy, and recognition of hazards imposed by previous surgery. Applying these principles, we describe a safe, rapid, and cost effective technique for laparoscopic access using readily available instruments. This technique emphasizes identification and incision of the point at which the midline abdominal fascia is fused with the base of the umbilicus, and the importance of the application of countertraction directly at the point of insertion. This method allows penetration under direct vision with minimal controlled axial force, and without the requirement for fascial sutures or other cumbersome aspects of the traditional open technique. While previous reports describe techniques for laparoscopic access entry based on similar anatomic and surgical principles, we describe an alternative method not yet discussed in the surgical literature. PMID- 15895195 TI - Outcomes after pylorus preserving gastrectomy for early gastric cancer. PMID- 15895196 TI - Functional and structural characterization of the minimal Sec translocase of the hyperthermophile Thermotoga maritima. AB - The genome of the hyperthermophilic bacterium Thermotoga maritima contains the genes that encode core subunits of the protein translocase, a complex consisting of the molecular motor SecA and the protein conducting pore SecYE. In addition, we identified an erroneous sequence in the genome encoding for a putative secG gene. The genes of the T. maritima translocase subunits were overexpressed in Escherichia coli and purified to homogeneity. T. maritima SecA showed a basal thermostable ATPase activity that was stimulated up to 4-fold by phospholipids with an optimum at 74 degrees C. Membrane vesicles and proteoliposomes containing SecYE or SecYEG supported 2- to 4-fold stimulation of the precursor dependent SecA ATPase activity. Imaging of small two-dimensional crystals of the SecYE complex using electron microscopy showed square-shaped particles with a side length of about 6 nm. These results demonstrate that in T. maritima a highly thermostable translocase complex is operational. PMID- 15895197 TI - Healthcare consumption and direct costs of rheumatoid arthritis in Belgium. AB - The aim of this study was to compare the socioeconomic consequences of early and late rheumatoid arthritis in Belgium and to assess the patient out-of-pocket contributions. This multicentre longitudinal study in Belgium evaluated patients with rheumatoid arthritis. Early disease was defined as diagnosis since less than 1 year. At baseline sociodemographic and disease characteristics were assessed and during the following year patients recorded all healthcare- and non healthcare-related direct costs and out-of-pocket contributions. The study included 48 patients with early and 85 patients with late rheumatoid arthritis. Mean disease duration was 0.5 vs 12.5 years in patients with early and late rheumatoid arthritis, respectively. The disease activity score (DAS28) was comparable between both groups (4.1 vs 4.5, p = 0.14), but physical function (Health Assessment Questionnaire, HAQ) was more impaired in patients with long standing disease (1.0 vs 1.7, p < 0.001). Work disability had increased from 2% in patients with early to 18% in patients with late disease. The annual societal direct costs per patient were 3055 Euros (median: 1518 Euros) opposed to 9946 Euros (median: 4017 Euros) for early and late rheumatoid arthritis, respectively. The higher direct cost for patients with long-standing disease was seen for all categories, but especially for physiotherapy and need for devices and adaptations. Patients with early as well as late disease contribute out of pocket about one-third to the direct healthcare costs. Within each group, HAQ was a strong determinant of costs. In Belgium, patients with long-standing rheumatoid arthritis are nine times more likely to be work disabled than patients with less than 1 year disease duration and have a threefold increase in costs. Differences in healthcare consumption between patients could be mainly explained by differences in physical function (HAQ). PMID- 15895198 TI - Significance of immunohistochemical expression of Rab3B and SNAP-25 in growth hormone-producing pituitary adenomas. AB - Exocytosis proteins play an important role in the secretory activities of anterior pituitary cells and adenoma cells. An immunohistochemical study was conducted to elucidate the functional significance of these proteins in growth hormone (GH)-producing adenomas. We studied 40 GH-producing adenomas, 10 prolactinomas, and 10 clinically nonfunctioning (NF) adenomas immunohistochemically with antibodies specific for Rab3B and SNAP-25, both of which are considered essential in secretory activities of the pituitary, and keratin 8 (clone CAM5.2). The tumor volume (TV) was estimated with high resolution magnetic resonance imaging. Immunoreactivity for Rab3B, which was granular in the cytoplasm, varied in GH adenomas, but was negative in prolactinomas and NF adenomas. Reactivity for SNAP-25, which was linear on the plasma membrane, varied in GH adenomas, and was intensely positive in prolactinomas, and negative in NF adenomas. In GH adenomas, an increased percentage of adenoma cells with dot-like immunoreactivity for keratin 8 was associated with decreased reactivities for Rab3B (R=0.739, P<0.0001) and SNAP-25 (R=0.840, P<0.0001). Increased reactivity for SNAP-25 correlated positively with plasma GH level per unit TV (R=0.685, P<0.0001). The immunoreactivities for Rab3B and SNAP-25 may reflect the number of secretory granules and exocytosis activity, respectively, in pituitary adenomas, including GH adenomas. PMID- 15895199 TI - [Dynamic decision making in emergency medicine. Example of paraplegia after a traffic accident]. AB - Dynamic decision making is one of the key skills in crew resource management training in aviation. In emergency medicine it is important to practice this skill as a prerequisite for effective treatment of patients. We report a case of paraplegia after a road traffic accident and cervical spine injury. During the prehospital treatment the patient's state was re-evaluated at different times. Although the patient was initially unconscious the physician at the scene decided not to intubate the trachea as the level of consciousness improved during resuscitation. In the emergency room a C5 fracture and a prolapsed intervertebral disc were diagnosed and immediate decompression and stabilisation of the cervical spine were performed. Dynamic decision-making has been in practise for a long time in aviation, similarities to decisions in medicine and the psychological background are described on the basis of the case report. PMID- 15895200 TI - [Preoperative abstinence from smoking. An outdated dogma in anaesthesia?]. AB - For decades it has been assumed, that smoking within 6 hours of anesthesia and surgery raises the incidence of perioperative cardiopulmonary complications (PPC) including aspiration. Therefore, every patient is advised to stop smoking at the day before surgery, and not to smoke at all at the day of surgery. If the patient does not follow this advice, this will result in a postponement of anesthesia and surgery. The present article aims at re-investigating the scientific basis of this dogma in anesthesia, which virtually forbids smoking at short-term prior to surgery. The influence of short-term (6 h) abstinence from smoking on the perioperative pulmonary morbidity has not been systematically investigated. Interestingly, giving up smoking less than two months prior to surgery does not significantly decrease, but rather may increase the incidence of PPC. With respect to the risk of aspiration, smoking does not increase either the volume or the acidity of gastric juices. A short-lived reduction in the tone of the lower esophageal sphincter is reversible within minutes after termination of smoking. While the emptying of liquid gastric juices is not influenced by smoking, there is a certain delay in the propulgation of solid food. This effect, however, is probably of no clinical relevance in patients, who had their last solid meal the evening before surgery. Hence, we conclude that the anesthesia dogma, which rules out smoking shortly prior to anesthesia, cannot be based on an otherwise increased incidence of pulmonary aspiration or other pulmonary morbidity. However, acute smoking (probably by an increase in COHb) may increase the incidence of myocardial ischemia during exercise and anesthesia. With reference to this possible cardiac complication it still seems reasonable to discourage smoking at least 12 to 48 hours prior to surgery in patients with elevated cardiac risk. PMID- 15895201 TI - Control of conformation changes associated with homologue recognition during meiosis. AB - During early meiosis, chromosomes pair via their telomeres and centromeres. This pairing induces a conformational change which propagates from these regions along each chromosome, making the chromatin of the partners accessible for intimate pairing. In the present study, we show by exploiting wheat-rye hybrids that the signal is initiated in both the presence and absence of either the Ph1 or Ph2 locus. However, the chromatin change only continues to propagate through rye telomeric heterochromatin when Ph1 is absent. This failure to propagate the chromatin change through the rye heterochromatin in the absence of Ph2 correlates with a subsequent lack of wheat-rye chromosome association at metaphase I. PMID- 15895202 TI - Identification of quantitative trait loci for race-nonspecific resistance to tan spot in wheat. AB - Tan spot, caused by Pyrenophora tritici-repentis (Ptr), is an economically important foliar disease in the major wheat growing areas throughout the world. Multiple races of the pathogen have been characterized based on their ability to cause necrosis and/or chlorosis on differential wheat lines. In this research, we evaluated a population of recombinant inbred lines derived from a cross between the common wheat varieties Grandin and BR34 for reaction to tan spot caused by Ptr races 1-3 and 5. Composite interval mapping revealed QTLs on the short arm of chromosome 1B and the long arm of chromosome 3B that were significantly associated with resistance to all four races. The effects of the two QTLs varied for the different races. The 1B QTL explained from 13% to 29% of the phenotypic variation, whereas the 3B QTL explained from 13% to 41% of the variation. Additional minor QTLs were detected but not associated with resistance to all races. The host-selective toxin Ptr ToxA, which is produced by races 1 and 2, was not a significant factor in the development of disease in this population. The race-nonspecific resistance derived from BR34 may take precedence over the gene for-gene interaction known to be associated with the wheat-Ptr system. PMID- 15895203 TI - Inheritance of field resistance to Stagonospora nodorum leaf and glume blotch and correlations with other morphological traits in hexaploid wheat (Triticum aestivum L.). AB - Breeding for wheat varieties resistant to Stagonospora nodorum blotch (SNB) is the most sustainable strategy for controlling the disease. In order to map quantitative trait loci (QTLs) for SNB resistance we analysed 204 recombinant inbred lines of the cross between the winter wheat (Triticum aestivum L.) variety Forno and the winter spelt (Triticum spelta L.) variety Oberkulmer. We determined the level of resistance of adult plants to leaf blotch (SNL) and glume blotch (SNG) as well as morphological traits for 2 years after artificial inoculation with S. nodorum. Using composite interval mapping and LOD > 3.7, we detected ten QTLs for SNG blotch resistance (six inherited from the susceptible parent Forno) and 11 QTLs for SNL resistance (four inherited from Forno) across 2 years. Both resistance traits were moderately correlated (r = 0.52) and had only one common QTL. For SNL resistance, seven QTLs were not associated with QTLs for morphological traits. Among them, QSnl.eth-2D, QSnl.eth-4B and QSnl.eth-7B3 had major effects (R(2) > 13%) and were potential candidates for marker-assisted selection. For SNG, the major QTL on chromosome 5A, explaining 36% of the phenotypic variance for resistance, was associated with the q locus conferring the spelt morphology (long lax ear, long culm and hard glumes). Only QSng.eth 1BS, which explained 7% of the variance for resistance to SNG blotch, was not associated with QTLs for morphological traits. The consequences for breeding programmes are discussed. PMID- 15895204 TI - Genealogical use of chloroplast DNA variation for intraspecific studies of Aegilops tauschii Coss. AB - Intraspecific patterns of chloroplast DNA variation was studied in Aegilops tauschii Coss., the D-genome progenitor of bread wheat. Nucleotide sequences of ten chloroplast microsatellite loci were analyzed for 63 accessions that cover the central part of the species distribution. As is often the case with nuclear microsatellites, those of chloroplasts of Ae. tauschii bear complex mutations. Several types of mutations other than change in the microsatellite repeat number were found, including base substitutions and length mutations in flanking regions. In total, eight mutations were present in the flanking regions of four loci. Most mutations in the flanking regions of microsatellite repeats are associated with biallelic polymorphisms. Phylogeographic analyses showed that such biallelic polymorphisms are useful to investigate intraspecific patterns of monophyletic lineage divergence. In contrast, most microsatellite repeat sites are multiallelic, variable within intraspecific lineages, and useful to compare degrees of genetic diversity between lineages. These findings show that the chloroplast genome harbors evolutionary variations informative for intraspecific studies of Ae. tauschii and can be analyzed by genealogical approaches. PMID- 15895206 TI - Anatomical analysis of the medial patellofemoral ligament of the knee, especially the femoral attachment. AB - The purpose of this study was to investigate the anatomical morphology and measurement of the medial patellofemoral ligament (MPFL), especially the femoral attachment. A total of 20 knee specimens were dissected and the total length, width, thickness, inclination, as well as the attachment points of the MPFL were measured. The MPFL was well-developed in seven knees, moderate in ten knees and wispy in three knees. Total length of the MPFL was 58.8 +/- 4.7 mm. The width and thickness was 12.0 +/- 3.1 mm and 0.44 +/- 0.19 mm at the middle point. The long axis of the MPFL inclined at 15.9 +/- 5.6 degrees proximally. The center of the patellar attachment was located at 27 +/- 10% from the upper end of the patella in the longitudinal patellar height. The femoral attachment was superoposterior to the medial femoral epicondyle and just distal to the adductor tubercle. The center of the anterior edge of the femoral attachment was 9.5 +/- 1.8 mm proximally and 5.0 +/- 1.7 mm posteriorly from the center of the medial femoral epicondyle. The femoral attachment was located at 61 +/- 4% of anteroposterior length of the medial femoral condyle from the anterior edge. PMID- 15895205 TI - Impact of selective decontamination of the digestive tract on fungal carriage and infection: systematic review of randomized controlled trials. AB - OBJECTIVE: To determine the impact of the antifungal component of selective decontamination of the digestive tract on fungal carriage, infection and fungaemia. DESIGN: Meta-analysis of randomized controlled trials of selective decontamination of the digestive tract. STUDY SELECTION: Data sources included Medline, Embase, Cochrane Register of Controlled Trials, previous meta-analyses, personal communications and conference proceedings, without restriction of language or publication status. All randomized trials were selected that compared oropharyngeal and/or intestinal administration of antifungals amphotericin B or nystatin, as part of selective decontamination protocol, with no treatment in the controls. There were 42 randomized controlled trials with a total of 6,075 critically ill patients. METHODS: Three reviewers independently applied selection criteria, performed quality assessment and extracted the data. The main outcome measures were patients with fungal carriage, patients with fungal infections and patients with fungaemia. Odds ratios were pooled with the random effect model. MEASUREMENTS AND RESULTS: Enteral antifungals significantly reduced fungal carriage (odds ratio 0.32, 95% confidence interval 0.19-0.53) and overall fungal infections (0.30, 0.17-0.53). Fungaemia was not significantly reduced in the treatment group (0.89, 0.16-4.95). CONCLUSIONS: Antifungals, as part of selective decontamination of the digestive tract, reduce fungal carriage and infection but not fungaemia in critically ill patients and may justify the inclusion of an antifungal component in the decontamination protocol. PMID- 15895207 TI - Short-term and long-term in vivo exposure to an ephedra- and caffeine-containing metabolic nutrition system does not induce cardiotoxicity in B6C3F1 mice. AB - Although conventional biomedical research has largely focused on mechanisms of weight loss and genetic aspects of obesity, most medical solutions are plagued by side-effects and fraught with complex questions. As a consequence, consumers are seriously considering herbal products, nutraceuticals and functional foods as alternatives to conventional medications. This is evidently driven by a growing consumer understanding of diet/disease links, aging-related consequences, rising health care costs, and advances in food technology and nutrition. This study investigated the effects of up to 12 months exposure to a multinutrient and botanical extract supplement (Metabolic Nutrition System Orange (MNSO) - sold by AdvoCare, Carrollton, TX, USA) at five dietary concentrations on serum biochemistry and target organ histopathology of the hearts of B6C3F1 mice. The MNSO is a unique combination of vitamins, minerals, omega-3 fatty acids and herbal extracts designed to provide a strong foundation of nutritional support, and to enhance thermogenesis and perception of energy. The MNSO contain extracts of citrus, ephedra, guarana, gingko, green tea and Ocimum. In this study, female B6C3F1 mice were fed control (-MNSO) or MNSO (one time to ten times, one time = daily human dose) diets. Animals were sacrificed after 4, 8 and 12 months', at which time blood was collected for serum chemistry analysis, and hearts were prepared for histopathology and tissue biochemistry. Food consumption and body weight changes were also monitored throughout the study. The MNSO exposure did not significantly affect any of the cardiosensitive enzymes [including creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST)] and normal histopathological architecture of the heart was observed. Although animals given the MNSO diet consumed more food, they were relatively leaner and more active compared to controls. The results indicate that ingestion of ephedra and caffeine for one year in the doses used as part of a comprehensive metabolic nutrition system does not significantly alter normal serum chemistry or induce any irreversible histological changes in the mouse heart, since this study employed up to ten times the normal human consumption dose of ephedra and the metabolic nutrition system. PMID- 15895209 TI - Chirally-functionalized monolithic materials for stereoselective capillary electrochromatography. PMID- 15895210 TI - Microgravimetric flow analysis of nucleic acid based on adsorption of nanoparticle-bioconjugate. AB - A novel nanoparticle-bioconjugate has been prepared by specific hybridization of the target with complementary thiol-labeled and nanoparticle-labeled probes. The rapid adsorption of the nanoparticle-bioconjugate onto a gold surface via a thiol gold reaction was monitored in real-time using a quartz crystal microbalance, and used to perform microgravimetric flow analysis of nucleic acid for the first time. This innovative assay is highly reproducible and sensitive, and shows great promise for clinical applications. PMID- 15895211 TI - Effluent analysis in analytical chemistry: an overview. AB - This review summarizes and discusses effluent analysis, focusing on the methods and techniques that have been most frequently described in the literature since 1975. The methods are classified into four main categories: (1) physical and chemical properties; (2) inorganic metals analysis; (3) inorganic non-metallic analysis; (4) organic analysis. PMID- 15895212 TI - The selectivity in MEKC of pseudo-stationary phases based on polyelectrolyte complexes: I. Composition of the complex. AB - Polyelectrolyte-surfactant complexes (PSC) of polycarboxylic acids with alkyl trimethylammonium salts look very promising as a new type of pseudo-stationary phase in micellar electrokinetic chromatography. PSC produce an intramolecular micellar phase, and the morphology of the micelles is significantly different from that of the corresponding "typical" surfactant micelles. Pseudo-stationary phases based on PSC have unique selectivity. In this paper, the effect of the composition (phi) of the PSC of polyacrylic acid (PAA) M(W) 130,000 with dodecyltrimethylammonium bromide (DTAB) and of the PSC of PAA M(W) 450,000 with DTAB on the separation of DNS-amino acids and phenol derivatives in these systems was investigated. Relative retention and relative selectivity were used to describe the electrophoretic behavior of the amino acids and phenol derivatives. The main advantage of PSC pseudo-phases is that the nature and the structure of micelle-like units, and hence the selectivity of electrophoretic separation, could easily be modified by changing the composition of the complex. PMID- 15895213 TI - The use of etched, chemically modified, rectangular capillaries as a separation medium for open tubular capillary electrochromatography. AB - The use of etched, chemically modified, capillaries with a rectangular inner channel for open tubular electrochromatography is investigated. Comparisons of separation capabilities are made between circular and rectangular capillaries undergoing the same etching and chemical modification processes. With the long dimension of the rectangular column aligned in the direction of the optical light path, the relative sensitivities of the two capillary geometries are evaluated. The electrochromatographic properties of two catechins found in tea are investigated on the rectangular etched octadecyl-modified capillary. PMID- 15895214 TI - Simultaneous determination of phenolic compounds by means of an automated voltammetric "electronic tongue". AB - This contribution describes the simultaneous determination of three phenolic compounds, o-cresol, p-chlorophenol and 4-chloro-3-methylphenol, using direct oxidation and amperometric detection coupled by signal deconvolution, accomplished via chemometric methods. Direct oxidation of phenolic compounds is performed at the surface of an epoxy-graphite transducer, by linear scan voltammetry. Due to strong signal overlapping, artificial neural networks (ANNs) were used during data treatment, in a combination of chemometrics and electrochemical sensors known as an "electronic tongue". To calibrate this system properly, a total of 80 mixed samples were prepared automatically by employing a sequential injection analysis (SIA) system designed to automatically generate the information needed to train the network. The phenolic compound concentration varied from 1 to 70 microM for o-cresol, from 0.5 microM to 140 microM for p chlorophenol and from 1 microM to 100 microM for 4-chloro-3-methylphenol. A good prediction capability was obtained, with correlation coefficients >0.964 when the obtained values were compared with those expected for a set of 24 external test samples not used for training. The results presented here indicate that this technique is a simple and robust analytical method of environmental interest. PMID- 15895215 TI - Detection and quantification of PAH in drinking water by front-face fluorimetry on a solid sorbent and PLS analysis. AB - The direct determination in drinking water of perylene, chrysene, pyrene, benzo[a]pyrene, and benzo[k]fluoranthene, by front-face synchronous fluorimetry on a commercial SPE disk, has been evaluated. Sorbent treatment, influence of humic substances, and pH effect are discussed. In pure water the detection limits were estimated to be in the range 0.03-0.01 microg L(-1). A working pH in the range 10-11 was found to minimize the fluorescence quenching effect of humic substances. The proposed method combined with a partial-least-square (PLS) treatment was tested for quantitative analysis of mixtures of four PAH in a spiked drinking water. PMID- 15895216 TI - Amperometric PMMA-microchip with integrated gold working electrode for enzyme assays. AB - The use of a gold film in-channel detector combined with a poly(methyl methacrylate) (PMMA) CE microchip has been tested for alkaline phosphatase (AP) enzymatic assays. Tris-borate or Tris-Gly (pH 9.0, 50 mmol L(-1)) buffer solutions were appropriate as running buffer. Signals for three common AP products: alpha-naphthol, p-nitrophenol, and ascorbic acid, were obtained. They were reproducible (RSD 4.4% for six successive electropherograms corresponding to 5 mmol L(-1) alpha-naphthol solution) and the response was dependent on concentration (linear relationship for ascorbic acid solutions between 5 and 20 mmol L(-1) concentration). Use of an end-channel gold film electrode was also investigated. If one of the reagents (substrate or enzyme) is included in the running buffer, two different types of enzymatic assay are feasible in less than 3 min. PMID- 15895217 TI - Human movement coordination implicates relative direction as the information for relative phase. AB - The current studies explore the informational basis of the coupling in human rhythmic movement coordination tasks. Movement stability in these tasks is an asymmetric U-shaped function of mean relative phase; 0 degrees is maximally stable, 90 degrees is maximally unstable and 180 degrees is intermediate. Bingham (2001, 2004a, 2004b) hypothesized that the information used to perform coordinated rhythmic movement is the relative direction of movement, the resolution of which is determined by relative speed. We used an experimental paradigm that entails using a circular movement to produce a linear motion of a dot on a screen, which must then be coordinated with a linearly moving computer controlled dot. This adds a component to the movement that is orthogonal to the display. Relative direction is not uniquely defined between orthogonal components of motion, but relative speed is; it was therefore predicted that the addition of the component would only introduce a symmetric noise component and not otherwise contribute to the U-shape structure of movement stability. Results for experiment 1 supported the hypothesis; movement that involved the additional component was overall less stable than movement that involved only the parallel component along which relative direction can be defined. Two additional studies ruled out alternative explanations for the pattern of data in experiment 1. Overall, the results strongly implicate relative direction as the information underlying performance in rhythmic movement coordination tasks. PMID- 15895218 TI - On the nature of the vestibular control of arm-reaching movements during whole body rotations. AB - Recent studies report efficient vestibular control of goal-directed arm movements during body motion. This contribution tested whether this control relies (a) on an updating process in which vestibular signals are used to update the perceived egocentric position of surrounding objects when body orientation changes, or (b) on a sensorimotor process, i.e. a transfer function between vestibular input and the arm motor output that preserves hand trajectory in space despite body rotation. Both processes were separately and specifically adapted. We then compared the respective influences of the adapted processes on the vestibular control of arm-reaching movements. The rationale was that if a given process underlies a given behavior, any adaptive modification of this process should give rise to observable modification of the behavior. The updating adaptation adapted the matching between vestibular input and perceived body displacement in the surrounding world. The sensorimotor adaptation adapted the matching between vestibular input and the arm motor output necessary to keep the hand fixed in space during body rotation. Only the sensorimotor adaptation significantly altered the vestibular control of arm-reaching movements. Our results therefore suggest that during passive self-motion, the vestibular control of arm-reaching movements essentially derives from a sensorimotor process by which arm motor output is modified on-line to preserve hand trajectory in space despite body displacement. In contrast, the updating process maintaining up-to-date the egocentric representation of visual space seems to contribute little to generating the required arm compensation during body rotations. PMID- 15895219 TI - White matter lesions in watershed territories studied with MRI and parenchymography: a comparative study. AB - Brain aging affects an increasing segment of the population and the role of chronic cerebrovascular disease is considered to be one of the main parameters involved. For this purpose we compared retrospectively MRI data with digitized subtraction angiography (DSA) data in a group of 50 patients focusing onto the watershed area of the carotid artery vascular territories. In order to evaluate the presence of white matter lesions (WML) in the hemispheric watershed areas, coronal fluid-attenuated inversion-recovery or axial T2 weighted MRI images of patients with symptomatic cerebrovascular insufficiency areas were compared with the capillary phase of DSA studies in anterior-posterior projection. Presence of cerebrovascular occlusive disease was evaluated on DSA using North American symptomatic carotid endarterectomy trial criteria and including evaluation of collateral vascular supply. Pathological MRI findings in the region of the watershed territories correlated overall in 66% of cases with a defect or delayed filling on DSA. In the case of asymmetrical MRI findings, there was a pathological finding of the capillary phase in the watershed area in 92% of DSA studies. Hypoperfusion in the capillary phase of the watershed area as seen on DSA correlated with the stenosis degree of the concerned carotid artery. Our findings suggest that asymmetrical findings of WML in the watershed areas as seen on MRI are caused by hemodynamic effect and a differentiation between small vessel disease and a consequence of distant stenosis may be possible under such conditions. PMID- 15895220 TI - Filling cystoureterography facilitates stone visualization in patients with previously inserted double-J ureteral stents. AB - In our study, we assessed the ability of a filling cystogram to induce vesicoureteral reflux (VUR) and to evaluate its role in stone targeting during ESWL in the presence of a double-J ureteral stent (JJ-S). Filling cystoureterography was performed during ESWL in 17 patients with previously inserted 4.7 F JJ-S. The examination ended with stone localization. In every procedure, 3,500 shock waves were delivered with an energy range of 18-23 kV. Patients were evaluated 4 weeks after ESWL. A stone-free result was defined as no evidence of calculi in the first or second visits. Because contrast material was employed in this study, we also evaluated its possible interference with the results of lithotripsy. All radiolucent or poorly calcified stones were successfully localized. In four patients, rhythmic suprapubic manual pressure was performed to initiate VUR. Reflux was low grade in 35% and high grade in 65% of renal units. The efficiency quotient reached 59%. In vitro artificial stones were successfully disintegrated both in water and solutions of Ultravist. In the presence of a JJ-S, filling cystography can easily localize radiolucent stones during SWL. No special catheters or stents are required for this technique. Ultravist in particular does not affect the results of ESWL unfavorably. PMID- 15895221 TI - A predictive model for the selective accumulation of chemicals in tumor cells. AB - Cationic lipophilic dyes can accumulate in mitochondria, and especially in mitochondria of tumor cells. We investigated the chemical properties and the processes allowing selective uptake into tumor cells using the Fick-Nernst-Planck equation. The model simulates uptake into cytoplasm and mitochondria and is valid for neutral molecules and ions, and thus also for weak electrolytes. The differential equation system was analytically solved for the steady-state and the dynamic case. The parameterization was for a generic human cell, with a 60 mV more negative potential at the inner mitochondrial membrane of generic tumor cells. The chemical input data were the lipophilicity (log K(OW)), the acid/base dissociation constant (pK(a)) and the electric charge (z). Accumulation in mitochondria occurred for polar acids with pK(a) between 5 and 9 owing to the ion trap, and for lipophilic bases with pK(a)>11 or permanent cations owing to electrical attraction. Selective accumulation in tumor cells was found for monovalent cations or strong bases with log K(OW) of the cation between -2 and 2, with the optimum near 0. The results are in agreement with experimental results for rhodamine 123, a series of cationic triarylmethane dyes, F16 and MKT-077, an anticancer drug targeting tumor mitochondria. PMID- 15895222 TI - Effect of environmental factors on the production of oxygenated unsaturated fatty acids from linoleic acids by Bacillus megaterium ALA2. AB - We identified many novel oxygenated fatty acids produced from linoleic acid by Bacillus megaterium ALA2: 12,13,17-trihydroxy-9(Z)-octadecenoic acid (12,13,17 THOA); 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA); 12-hydroxy 13,16-epoxy-9(Z)-octadecenoic acid; and 12,17;13,17-diepoxy-16-hydroxy-9(Z) octadecenoic acid. 12,13,17-THOA, the main product, has antiplant pathogenic fungal activity. To develop an industrial process for the production of these new oxygenated fatty acids by strain ALA2, the effect of environmental factors on the production and their impact on the amount of various products were studied. Dextrose at 5 g/l was the optimum amount for the carbon source. A combination of 15 g yeast extract and 10 g tryptone showed good results as nitrogen sources. Among the metal ions tested, the optimum concentrations for the reaction for the different ions were as follows (in mM): magnesium 2.0, iron 0.5, zinc 0.1, nickel 0.01, and cobalt 0.05. Copper ions did not affect the production of oxygenated products; however, manganese ions inhibited the reaction. Addition of these metal ions did not alter the distribution of products. The optimum temperature and pH for the production of THOAs were 30 degrees C and pH 6.5. Time course studies showed 40-48 h is the optimum for the production of both THOAs. These data provide the basis for engineering scale-up production of these new products. PMID- 15895223 TI - The quadruple cruciate sign of simultaneous bicompartmental medial and lateral bucket-handle meniscal tears. AB - We report the second known case of bicompartmental bucket-handle tears of the medial and lateral menisci and the first documented case of the bucket-handle tears occurring simultaneously following trauma, which occurred after a motorcycle accident. Both bucket-handle fragments were displaced into the intercondylar notch. An anterior cruciate ligament tear was also present. Coronal images demonstrated four structures in the intercondylar notch: the anterior and posterior cruciate ligaments and the medial and lateral bucket-handle meniscal fragments. PMID- 15895224 TI - Rosai-Dorfman disease presenting as multiple soft tissue masses. AB - Rosai-Dorfman disease is a rare clinical disorder which may present in many forms. While classically a disease of lymph nodes, soft tissue involvement is fairly common. Soft tissue involvement can occur without any lymphatic or systemic involvement, and may be difficult to diagnose. We describe a patient presenting with multiple soft tissue masses which on biopsy proved to be isolated cutaneous Rosai-Dorfman disease. MR imaging showed two well-defined nonspecific superficial masses that enhanced intensely. Review of the literature suggests that when this disease presents in soft tissue, multiple foci of involvement may be common. Although rare, Rosai-Dorfman disease should be considered in the differential diagnosis of patients presenting with multiple soft tissue masses. PMID- 15895225 TI - Kimura's disease of the elbows. AB - Kimura's disease rarely involves a location outside the head and neck. We report a case of Kimura's disease in a young Asian man involving both elbows. Magnetic resonance imaging showed a soft-tissue mass of irregularly infiltrative strands in the subcutaneous fat accompanied with medial epitrochlear lymphadenopathy. The MRI appearance is described and the entity is briefly reviewed. PMID- 15895226 TI - Evaluation of posterior tibial pathology: comparison of sonography and MR imaging. AB - OBJECTIVE: To compare the results of sonographic (US) and magnetic resonance (MR) imaging in detecting pathology of the posterior tibial tendon (PTT) in patients with PTT dysfunction. DESIGN: Twenty-two ankles that were clinically suspected by the orthopedic surgeon to have PTT dysfunction were evaluated with US (10 MHz linear-array transducer) and 1.5 T MR examinations within the same day. The US and MR studies were conducted and interpreted by two sonologists and two musculoskeletal radiologists who were masked to the results of the other study. Four patients had bilateral studies. Classic clinical findings were utilized as a standard reference in staging PTT dysfunction. PATIENTS: Eighteen women (mean age 61 years, age range 39-86 years). RESULTS: Based on a commonly accepted staging system for PTT dysfunction, 6 ankles were classified as stage I, 11 ankles as stage II, and 5 ankles as stage III. All stage I ankles were interpreted as having an intact PTT by both MR imaging and US. In the stage II and III tendons, MR imaging demonstrated PTT tears in 12 of 22 examinations, including 11 partial tears and 1 complete tear. US demonstrated PTT tears in 8 of 22 examinations, including 8 partial tears and no complete tears. The findings of US and MR imaging were consistent in 17 of 22 cases (77%). The five inconsistencies were as follows: in 4 cases, US reported tendinosis when MR imaging interpreted partial tears (no change in management); in one case, US diagnosed a partial tear when MR reported a complete tear of the PTT (no change in management because the clinical findings were more consistent with a partial tear). CONCLUSIONS: In this study, US and MR imaging of the PTT were concordant in the majority of cases. US was slightly less sensitive than MR imaging for PTT pathology, but these discrepancies did not affect clinical management. PMID- 15895227 TI - FDG imaging should be considered the preferred technique for accurate assessment of myocardial viability: against. PMID- 15895228 TI - 67Ga-SPECT/CT with a hybrid system in the clinical management of lymphoma. AB - PURPOSE: The purpose of this study was to investigate the added value of co registered fusion imaging using a hybrid system in patients with lymphoma. METHODS: Twenty-four lymphoma patients underwent (67)Ga-SPECT/CT using a hybrid tomograph consisting of a dual-head, variable-angle gamma camera and a low-dose X ray tube. Results were compared with those of SPECT alone. RESULTS: Forty-five lesions were identified by SPECT alone, while 49 were detected by SPECT/CT. Forty out of the 45 lesions observed on SPECT were confirmed as lymphoma, but five were due to other causes (thoracic aorta blood pool activity, sialoadenitis in the submandibular gland, bowel activity, rib fracture and bone marrow activation due to radiotherapy). SPECT/CT identified nine more neoplastic lesions compared with SPECT alone: four areas of radiopharmaceutical accumulation were observed in para aortic lymph nodes, three in the spleen, one in the liver and one in para-iliac lymph nodes. In five cases, SPECT/CT provided additional anatomical information over SPECT alone. In four patients, four large areas of (67)Ga uptake (one mediastinal, two supraclavicular and one para-aortic) were better characterised; in one subject uptake was localised in the seventh thoracic vertebra only by SPECT/CT. Hybrid imaging provided additional data in 13 patients (54.2%), thus inducing oncologists to reconsider the therapeutic approach in eight subjects (33.2%): unnecessary treatment was avoided in four (16.6%) while therapy was altered in another four (16.6%). CONCLUSION: SPECT/CT hybrid system is able to provide information not obtained by SPECT alone. It allows the anatomical localisation of lymphoma and physiological radiopharmaceutical uptake, facilitates the diagnosis of tumours located in the abdomen (subdiaphragmatic lesions) and provides information that may cause a change in therapeutic strategy. PMID- 15895229 TI - Hemolytic uremic syndrome following prolonged gemcitabine therapy: report of four cases from a single institution. Muller S, Schutt P, Bojko P, Nowrousian MR, Hense J, Seeber S, Moritz T. Ann. Hematol. 2005 Feb; 84(2):110-4. PMID- 15895230 TI - Modulation of irinotecan with cyclosporine: a phase II trial in advanced colorectal cancer. AB - INTRODUCTION: Despite the extensive clinical experience with irinotecan, significant concerns remain regarding its toxicity. In a phase I trial, we modulated irinotecan pharmacokinetics by inhibiting biliary excretion of SN-38, the active metabolite of irinotecan, using cyclosporine. The modulation appeared to decrease the gastrointestinal toxicity of irinotecan and suggested that irinotecan activity might also be retained. Hence, we conducted this phase II trial in patients with colorectal cancer (CRC) to further evaluate the toxicity and activity of irinotecan modulated with cyclosporine. PATIENTS AND METHODS: Sixteen patients with 5-fluorouracil refractory CRC were treated. Cyclosporine (5 mg/kg) was administered as a 6-h infusion and irinotecan (60 mg/m2/day, 90-min infusion) was started 3 h after initiation of the Cyclosporine. Both agents were given weekly for 4 weeks, every 6 weeks. Responses were assessed every 12 weeks, and toxicity was monitored weekly. RESULTS: Sixteen patients were evaluable for toxicity and 11 for response. There was 1 partial response (6%). Five patients had SD lasting a median of 12 weeks. Grade 3/4 diarrhea was observed in only 13% of the patients. CONCLUSION: Pharmacokinetic modulation of irinotecan using parenteral cyclosporine appears to decrease the incidence of diarrhea in CRC patients. Given the modest activity of irinotecan monotherapy, a larger study would be required to assess if the modulation improves the toxicity without compromising this activity. The available clinical data suggest that pharmacokinetic modulation of irinotecan should be evaluated further to define its optimal clinical utility. PMID- 15895231 TI - Augmenting tumor sensitivity to topotecan by transient hypoxia. AB - We examined how the effect of topotecan is modulated by transient hypoxia in three different tumor lines, Lewis lung carcinoma (LLC), U87 human glioblastoma and DMS273 human small cell lung cancer. Four groups of tumor bearing mice were treated with saline or a single dose of topotecan, immediately followed by 6-h or 72-h exposure to a hypoxic environment (10% O(2)) or normal air. Topotecan + hypoxia resulted in significantly greater suppression of tumor growth than normoxic topotecan or hypoxia alone. Correspondingly, the sensitivity of LLC cells to topotecan in a clonogenic survival assay was significantly higher under hypoxia. This effect of hypoxia was not a general phenomenon, since the tumor growth inhibitory effect of the alkylating agent cisplatin was not changed by hypoxic environment. In a parallel series of in vitro experiments, cell cultures were exposed to hypoxia (0.1% or 0.7% O(2)) in a hypoxic chamber or normoxia for 24 h. We found a dose-dependent downregulation of HIF-1alpha by topotecan (30-270 nM). The hypoxic upregulation of Glucose transporter-1 and VEGF secretion to the culture medium was inhibited by the addition of topotecan, while doses up to 270 nM had no effect on VEGF under normoxia. VEGF protein levels in tumors were also reduced by topotecan. These data show that the effect of topotecan is increased by transient hypoxia, and this may be a direct effect on the ability of cells to survive under hypoxia as well as an antiangiogenic effect, mediated through the HIF-1 inhibitory effect of topotecan. PMID- 15895232 TI - Platinum-acridinylthiourea conjugates show cell line-specific cytotoxic enhancement in H460 lung carcinoma cells compared to cisplatin. AB - Recently, we reported a new class of DNA-targeted hybrid platinum-acridine agents. The parent intercalator, ACRAMTU, a 9-aminoacridine derivative, intercalates into the minor groove of DNA, causing the corresponding prototypical conjugate, PT-ACRAMTU (type I/n=2), to form DNA adducts dissimilar to traditional platinum drugs. Both these agents show cytotoxic activity in leukemic and ovarian cancer cells. Following the use of clonogenic survival assays, we report on the cytotoxic effects of ACRAMTU, PT-ACRAMTU, and three PT-ACRAMTU derivatives, on additional cell lines including colon (RKO), lung (H460), and cisplatin-sensitive (A2780) and cisplatin-resistant (A2780/CP) ovarian cells. While a dose-dependent effect was observed with both ACRAMTU and PT-ACRAMTU, an enhanced cytotoxic effect was seen with PT-ACRAMTU in all cell lines. PT-ACRAMTU appeared to have a similar IC50 value to cisplatin except in H460 lung cancer cells in which PT ACRAMTU had a twofold lower IC50 value. PT-ACRAMTU appeared to act in a time dependent manner. In H460 cells the IC50 value of PT-ACRAMTU was 235-fold higher following a 1-h incubation than following a 24-h incubation (0.27 microM), while following an 8-h incubation the IC50 value was 0.41 microM. Three derivatives of PT-ACRAMTU were also tested. A tetraalkylated derivative, type II/n=2, generated the highest IC50 values in all cell lines, while the trialkylated derivative, type III/n=2, generated IC50 values similar to its isomer, PT-ACRAMTU. PT-ACRAMTU with an added CH2 group in the thiourea linker (type I/n=3) showed IC50 values similar to the type I/n=2 prototype in H460 lung cells. An apoptotic response to PT-ACRAMTU appeared to be generated in H460 cells as evidenced by DNA laddering. These results suggest that type I/n=2 and type I/n=3 may be promising agents for the treatment of lung cancer and should be pursued in animal models. PMID- 15895233 TI - Metabolic profile of XK469 (2(R)-[4-(7-chloro-2-quinoxalinyl)oxyphenoxy] propionic acid; NSC698215) in patients and in vitro: low potential for active or toxic metabolites or for drug-drug interactions. AB - As part of an ongoing phase 1 study, we studied the excretion of XK469 and its metabolism in patients and in vitro. Five primary metabolites were identified by HPLC/MS/MS. An oxidized product formed by cytosolic aldehyde oxidase was the predominant species both in urine and human hepatocytes in vitro. Conjugates of XK469 with glycine, taurine, and glucuronic acid, as well as the microsomal product, 4-oxo-XK469, were also found in urine and in vitro, but none were major contributors to the mass balance for XK469 elimination. Based upon the relative concentrations circulating in plasma, systemic exposure to parent drug was 100 fold higher than for the metabolites. Thus, both toxicity and efficacy of XK469 are most likely to be produced by the parent molecule, rather than the metabolites. Urinary recovery of parent drug was low (2% of dose in 24 h), partly because of the long half-life of XK469 (approximately 3 days). In addition, the metabolite profile in urine indicates that only 25% of the XK469-derived material was unchanged drug. Thus, urinary excretion was not a major factor in XK469 elimination. Variations in systemic exposure to XK469 will be strongly influenced by factors that alter the activity of aldehyde oxidase, including pharmacogenetics, enzyme inhibition, and enzyme induction, but no specific modifiers have been reported. The multiday half-life of XK469 hampered our ability to obtain a complete mass balance, and the possibility exists that other routes, such as biliary excretion, may also play a substantial role in XK469 disposition. PMID- 15895234 TI - Small-bowel intussusception as a rare differential diagnosis in HIV-positive patients with acute abdominal pain. PMID- 15895235 TI - Whole-body magnetic resonance angiography of patients using a standard clinical scanner. AB - The purpose of this study was to evaluate the technique of whole-body magnetic resonance angiography (MRA) of patients with a standard clinical scanner. Thirty three patients referred for stenoses, occlusions, aneurysms, assessment of patency of vascular grafts, vasculitis and vascular aplasia were examined in a 1.5-T scanner using its standard body coil. Three-dimensional sequences were acquired in four stations after administration of one intravenous injection of 40 ml conventional gadolinium contrast agent. Different vessel segments were evaluated as either diagnostic or nondiagnostic and regarding the presence of stenoses with more than 50% diameter reduction, occlusions or aneurysms. Of 923 vessel segments, 67 were not evaluable because of poor contrast filling (n=31), motion artefacts (n=20), venous overlap (n=12) and other reasons (n=4). Stenoses of more than 50%, occlusions or aneurysms were observed in 26 patients (129 segments). In nine patients additional unsuspected pathology was found. In 10 out of 14 patients (71/79 segments) there was conformity between MRA and digital subtraction angiography regarding the grade of stenosis. This study shows that whole-body MRA with a standard clinical scanner is feasible. Motion artefacts and the timing of the contrast agent through the different segments are still problems to be solved. PMID- 15895236 TI - [Influence of intraocular lens material and design on the development of posterior capsule opacification]. AB - Based on the literature of 1999-2005, we attempted to clarify the influence of material and design of an intraocular lens (IOL) on the development of posterior capsule opacification (PCO) and identify the future problems to be solved. The sharp edge design of an IOL has significantly reduced PCO in recent years. Histopathological studies showed that a sharp capsular bend was formed at the sharp posterior optic edge, and the migration of lens epithelial cells was obviously inhibited at the site. Experimental and clinical studies suggest that the sharper the edge is, the sharper the capsular bend, and the greater the preventive effect, regardless of IOL material. The sharp capsule bend appears to represent a physical hindrance, which may induce contact inhibition of cell movement. How material participates in the preventive effect should be clarified in a future study. Thus, at the moment, the formation of capsular bend at the posterior optic edge appears to be the decisive criterion to evaluate the influence of IOL material and design on the development of PCO. Optimization of design and material of an IOL, which facilitates capsular bend formation as sharply and quickly as possible, may reduce PCO to a clinically negligible level. PMID- 15895237 TI - Spatial memory and orientation strategies in the elasmobranch Potamotrygon motoro. AB - We investigated whether juvenile freshwater stingrays (Potamotrygon motoro) can solve spatial tasks by constructing a cognitive map of their environment. Two experimental conditions were run: allocentric and ego-allocentric. Rays were trained to locate food within a four-arm maze placed in a room with visual spatial cues. The feeding location (goal) within the maze (room) remained constant while the starting position varied for the allocentrically but not for the ego-allocentrically trained group. After training, all rays solved the experimental tasks; however, different orientation strategies were used within and between groups. Allocentrically trained rays reached the goal via novel routes starting from unfamiliar locations, while ego-allocentrically trained rays primarily solved the task on the basis of an egocentric turn response. Our data suggest that P. motoro orients by constructing a visual cognitive map of its environment, but also uses egocentric and/or other orientation strategies alone or in combination for spatial orientation, a choice which may be governed by the complexity of the problem. We conclude that spatial memory functions are a general feature of the vertebrate brain. PMID- 15895238 TI - Poly (adp-ribose) synthetase inhibition reduces oxidative and nitrosative organ damage after thermal injury. AB - Poly (ADP-ribose) synthetase (PARS) is a nuclear enzyme activated by DNA single strand breakage, which can be triggered by reactive oxygen and nitrogen species. Activation of this enzyme depletes the intracellular concentration of energetic substrates such as nicotinamide adenine dinucleotide (NAD). Eventually, this process results in cell dysfunction and cell death. PARS inhibitors have successfully shown benefits in several experimental models of ischemia reperfusion injury, inflammation, and sepsis. In our experimental study, we investigated the role of 3-aminobenzamide (3-AB), a nonspecific PARS inhibitor, in systemic organ damage after burn. Twenty-four Wistar rats were randomly divided into three groups. The sham group (n=8) was exposed to 21 degrees C water, and the burn group (n=8) and the burn-plus-3-AB group (n=8) were exposed to boiling water for 12 s to produce a full-thickness burn of 35-40% of total body surface area. In the burn-plus-3-AB group, 3-AB 10 mg/kg was given intraperitoneally 10 min before thermal injury. Twenty-four hours later, tissue samples were obtained for biochemical analysis from lung, intestine, and kidney. In the burn group, tissue malondialdehyde, myeloperoxidase, and 3-nitrotyrosine levels in all organs were significantly increased compared with the sham group (p<0.05). Pretreatment with 3-AB significantly reduced burn-induced organ damage (p<0.05). These data provide evidence of the relationship between the PARS pathway and lipid peroxidation in systemic organ damage after thermal injury. PMID- 15895239 TI - Mitogen-activated protein kinases are developmentally regulated during stress induced microspore embryogenesis in Brassica napus L. AB - Plant mitogen-activated protein kinase (MAPK) cascades are involved in extracellular stress signalling pathways, leading to different cellular responses. Stress-induced microspore embryogenesis involves the internalization of an extracellular stress signal, generating a number of cellular responses where MAPK cascades might be involved. These responses include a change of the developmental programme, the entry into an early proliferative stage and, subsequently, into differentiation stages during haploid embryogenesis. In this work we studied the expression during microspore embryogenesis of several kinases, to assess their putative role in these events. The known Brassica napus MAP kinase kinase kinases (MAP3Ks BnMAP3Kalpha1, BnMAP3Kbeta1 and BnMAP3Kepsilon, the BnBSKtheta kinase and B. napus extracellular signal-regulated kinase (ERK) homologues were analysed by electron microscope (EM) in situ hybridization, immuno-gold labelling, immunofluorescence and western blotting. The differential in situ expression of these kinases suggests a role for them during embryogenesis. Two different expression patterns were observed, indicating a different regulation. BnMAP3Kalpha1, BnMAP3Kepsilon, and the ERKs showed a pattern consistent with a role mainly in proliferative events. Conversely, BnMAP3Kbeta1 and BnBSKtheta, presented a pattern that suggested an involvement in differentiation stages. In addition, ERK homologues migrate to the nucleus immediately after induction, being found in a phosphorylated state in a larger amount. PMID- 15895240 TI - Transient upregulation of connexin43 gap junctions and synchronized cell cycle control precede myoblast fusion in regenerating skeletal muscle in vivo. AB - The spatio-temporal expression of gap junction connexins (Cx) was investigated and correlated with the progression of cell cycle control in regenerating soleus muscle of Wistar rats. Notexin caused a selective myonecrosis followed by the complete recapitulation of muscle differentiation in vivo, including the activation, commitment, proliferation, differentiation and fusion of myogenic cells. In regenerating skeletal muscle, only Cx43 protein, out of Cx-s 26, -32, 37, -40, -43 and -45, was detected in desmin positive cells. Early expression of Cx43 in the proliferating single myogenic progenitors was followed by a progressive upregulation in interacting myoblasts until syncytial fusion, and then by a rapid decline in multinucleate myotubes. The significant upregulation of Cx43 gap junctions in aligned myoblasts preceding fusion was accompanied by the widespread nuclear expression of cyclin-dependent kinase inhibitors p21(waf1/Cip1) and p27(kip1) and the complete loss of Ki67 protein. The synchronized exit of myoblasts from the cell cycle following extensive gap junction formation suggests a role for Cx43 channels in the regulation of cell cycle control. The potential of Cx43 channels to stimulate p21(waf1/Cip1) and p27(kip1) is known. In the muscle, proving the involvement of Cx43 in either a direct or a bystander cell cycle regulation requires functional investigations. PMID- 15895241 TI - Expression of the potassium channel ROMK in adult and fetal human kidney. AB - The renal potassium channel ROMK is a crucial element of K+ recycling and secretion in the distal tubule and the collecting duct system. Mutations in the ROMK gene (KCNJ1) lead to hyperprostaglandin E syndrome/antenatal Bartter syndrome, a life-threatening hypokalemic disorder of the newborn. The localization of ROMK channel protein, however, remains unknown in humans. We generated an affinity-purified specific polyclonal anti-ROMK antibody raised against a C-terminal peptide of human ROMK. Immunoblotting revealed a 45 kDa protein band in both rat and human kidney tissue. In human kidney sections, the antibody showed intense staining of epithelial cells in the cortical and medullary thick ascending limb (TAL), the connecting tubule, and the collecting duct. Moreover, a strong expression of ROMK protein was detected in cells of the macula densa. In epithelial cells of the TAL expression of ROMK protein was mainly restricted to the apical membrane. In human fetal kidney expression of ROMK protein was detected mainly in distal tubules of mature nephrons but not or only marginally in the collecting system. No expression was found in early developmental stages such as comma or S shapes, indicating a differentiation dependent expression of ROMK protein. In summary, these findings support the proposed role of ROMK channels in potassium recycling and in the regulation of K+ secretion and present a rationale for the phenotype observed in patients with ROMK deficiency. PMID- 15895242 TI - Employment status and quality of life in patients with chronic obstructive pulmonary disease. AB - OBJECTIVES: Recently, the conditions for disability benefits were redefined in several countries, stimulating employees to participate on the labour market as long as reasonably possible. Little is known of labour participation and quality of life (QoL) of employees with chronic diseases. This study examines the associations between employment status and QoL in COPD patients. Additionally, the role of lung function, and work-related symptoms and exposures on QoL were explored. METHODS: Secondary analyses were conducted on baseline data from a randomized control trail. Patients were categorized as: "paid-workers"; "voluntary non-paid workers" (e.g., early retired, house wives) or "disabled for work". QoL was assessed by the Chronic Respiratory Questionnaire (CRQ). Between group differences in CRQ scores, and associations between work-related symptoms or irritants and CRQ scores were uni- and multivariately analysed. RESULTS: Two hundred and ten patients with COPD were included [mean age 53.9 (SD 6.8) years, FEV(1) %predicted 63.5 (SD 18.5)]. No statistically significant differences in lung function between the employment status subgroups were observed. Multivariable analysis showed that the "disabled" had lower CRQ scores as compared with "paid workers" (0.52 point difference, p<0.001). The CRQ scores of "voluntary non-paid workers" were not significantly different from "paid workers". Within the group paid workers, patients with many work-related clinical symptoms and being susceptible to various work-related irritants experienced a lower QoL than patients who had respectively no symptoms, or who were not susceptible to these factors. CONCLUSIONS: Patients with COPD who were disabled for work showed equal severity of airflow limitation but worse QoL, as compared with paid workers. PMID- 15895243 TI - Nephrotoxicity of organic solvents: biomarkers for early detection. AB - OBJECTIVES: Evidence for a relationship between chronic kidney diseases or progression of already existing diseases (glomerulonephritides) and occupational solvent exposure has been found in case reports, in case-control studies and also in cross-sectional studies. An analysis of the available literature was performed with respect to markers measured in cross-sectional studies that might be useful for an early detection of solvent-induced effects on the kidney. METHODS: The relevant cross-sectional studies were evaluated and the following markers were analyzed with respect to their suitability as biomarker for renal damage: total protein, albumin, transferrin, IgG, beta(2)-microglobulin, retinol-binding protein, N-acetyl-beta-D: -glucosaminidase, alanine aminopeptidase, beta galactosidase, beta-glucuronidase, leucin aminopeptidase, alkaline phosphatase, lysozyme, Tamm-Horsfall protein and laminin fragments in urine as well as E selectin, laminin and anti-laminin antibodies and anti-glomerular basement membrane antibodies in serum. RESULTS: An increased albumin excretion was observed more frequently in groups of workers exposed to various solvents (like toluene, styrene, aliphatic/aromatic hydrocarbon mixtures, tetrachloroethene, mixtures of chlorinated hydrocarbons) than in controls. No clear pattern emerged for the other markers. CONCLUSIONS: The determination of albumin excretion in the urine appears to be a useful parameter for monitoring solvent-exposed workers. PMID- 15895244 TI - Neurobehavioral science in hazard identification and risk assessment of neurotoxic agents--what are the requirements for further development? AB - OBJECTIVES: Modern neurobehavioral methods find useful application in research into the early effects of exposure to neurotoxic agents in the environment. This paper briefly describes the history and evolution of neurobehavioral toxicology, reviews some current trends in research in this specific discipline and identifies the most important needs and challenges to be addressed in future studies. METHODS: All published literature was considered, including ad hoc meeting reports. Further information was obtained directly from experts in the field. RESULTS: The number of studies (including those in occupational, environmental and pediatric exposure) using neurobehavioral evaluation is constantly increasing. Regulatory agencies are using scientific data obtained through neurobehavioral assessment, which includes other areas such as neurosensory toxicology. However, further development of this discipline is facing a number of problems and issues. Three major areas that deserve further attention have been identified: (1) specific technical issues regarding testing development, (2) epidemiological issues regarding the study design, including the need for meta-analysis/multi-center studies and for longitudinal observation, and statistical issues regarding the most adequate models for the analysis and treatment of complex neurobehavioral datasets, and (3) the need for scientific consensus on the significance of adverse effects identified with neurobehavioral methods. CONCLUSIONS: The importance of neurobehavioral toxicology in the evaluation of mechanisms of action and for preventive purposes is progressively growing. Further development is needed for the advancement of this discipline through collaboration between experts from different fields. PMID- 15895245 TI - Respiratory symptoms and diseases among construction painters. AB - OBJECTIVE: To study the self-reported prevalence of respiratory symptoms and diseases among construction painters and estimate the potential risk for this group compared with a representative group of carpenters sharing the construction work environment but without significant exposure to paint. METHODS: A questionnaire study was conducted on 1,000 male Finnish construction painters and 1,000 carpenters (mean response rate 60.5%). Symptoms and diseases of the respiratory tract were studied, by logistic regression modelling, in relation to occupation and duration of painting experience. Age, atopy and smoking habits were taken into account. RESULTS: The painters reported more asthma-like, rhinitis, laryngeal and eye symptoms than the carpenters [odds ratio (OR) 1.4 1.8]. The difference in the prevalence of asthma between the occupations was not statistically significant, but the painters with 1-10 years of painting experience had a threefold risk of asthma compared with the carpenters. Chronic bronchitis was linked to painting occupation [OR 1.9, 95% confidence interval (CI) 1.2-3.0] and to the duration of exposure; OR (CI) for over 30 years of painting was 2.2 (1.2-4.0). Occupation was not associated with allergic rhinitis or conjunctivitis. CONCLUSIONS: The results indicate a higher risk for respiratory symptoms and chronic bronchitis among construction painters than among carpenters. PMID- 15895246 TI - The mammalian melastatin-related transient receptor potential cation channels: an overview. AB - The mammalian melastatin-related transient receptor potential (TRPM) subfamily contains eight members. TRPM proteins, consisting of six putative transmembrane domains and intracellular N and C termini, form monovalent-permeable cation channels with variable selectivity for Ca(2+), Mg(2+) and other divalent cations. Some functions are linked to their individual cation selectivity: the highly divalent-permeable cation channels TRPM6 and TRPM7 are involved in the control of Mg(2+) influx, whereas the Ca(2+)-impermeable channels TRPM4 and TRPM5 modulate cellular Ca(2+) entry by determining the membrane potential. TRPM2, TRPM3 and TRPM8 mediate a direct influx of Ca(2+) in response to specific stimuli. Electrophysiological properties of the founding member, melastatin (TRPM1), are unexplored. The individual TRPM members are activated by different stimuli, including voltage, Ca(2+), temperature, cell swelling, lipid compounds and other endogenous or exogenous ligands. This review summarizes molecular features, activation mechanisms, biophysical properties and modulators of TRPM channels. PMID- 15895247 TI - Inhibition of TRPC5 channels by Ca2+-binding protein 1 in Xenopus oocytes. AB - The transient receptor potential canonical type 5 (TRPC5) channel is a member of the channels that has been implicated in neurite extension and growth cone morphology of hippocampal neurons. Although homomeric TRPC5 channels are activated following stimulation of G(q/11)-coupled receptors, the exact mechanism for this activation remains unresolved. Using two-electrode voltage clamp recordings, we show that the activity of TRPC5 channels expressed in Xenopus oocytes is dependent on the presence of Ca2+ at the extracellular as well as the cytoplasmic side of the plasma membrane. TRPC5 was activated by the stimulation of coexpressed M5 muscarinic receptors or by ionomycin. The TRPC5 activity was detectable with the presence of submillimolar levels of extracellular Ca2+, but it was eliminated by the injection of 5 mM 1,2-bis(o-aminophenoxy)ethane N,N,N',N'-tetraacetic acid into the oocytes. Lanthanum could substitute for extracellular Ca2+ to support TRPC5 activity. Coexpression of Ca2+-binding protein 1 (CaBP1), but not calmodulin (CaM), inhibited the TRPC5 activity, without affecting the cell surface expression of TRPC5 proteins. Using in vitro binding assays, we demonstrated direction interactions between CaBP1 and TRPC5. The CaBP1-binding sites at the C terminus of TRPC5 are closely localized, but not identical, to CaM-binding sites. We conclude that TRPC5 is a Ca2+-regulated channel, and its activity is negatively controlled by CaBP1. PMID- 15895248 TI - Subcellular localization and trafficking of polycystins. AB - Polycystin-2 is a member of the transient receptor potential (TRP) family of ion channels that is mutated in autosomal dominant polycystic kidney disease. Although its function as a non-selective cation channel has been demonstrated in several model systems, the precise subcellular localization of polycystin-2 (TRPP2) in tubular epithelial cells has remained controversial. Recent evidence suggests that the subcellular localization of TRPP2 is regulated by multiple protein interactions. This review will summarize our current knowledge about polycystin trafficking and highlight the experimental data that supports a compartment-specific function of 'cystogenic' proteins. PMID- 15895249 TI - Immunocytochemical characterization of the incubated rat renal cortical slices. AB - The use of renal cortical slices in vitro and the data obtained in these studies have been subjects of controversy, largely due to uncertain viability, e.g., structural and functional integrity of the proximal and other tubules. However, detailed studies of tubule integrity have not been reported. To correlate functional and structural viability of the hand-cut rat renal cortical slices, incubated in optimally conditioned media for up to 25 h, we studied the time course of p-aminohippurate (PAH) uptake, the immunocytochemical distribution of several proteins that reside in the proximal tubule basolateral [Na/K-ATPase, organic anion transporters (OAT)1 and OAT3], or brush border [megalin, sodium proton exchanger (NHE)3] membrane, as well as the general integrity of the tubule epithelium and its cytoskeleton (actin filaments, microtubules). PAH uptake in slices was proportional to time within 1 h of incubation and gradually declined thereafter. The immunostaining experiments indicated a fast, time-dependent loss of basolateral transporters, at a rate of OAT1 > Na/K-ATPase > OAT3. In the brush border membrane, the loss of megalin was faster than that of NHE3, and a partial redistribution of NHE3 into the basolateral domain indicated the loss of cell polarity. The loss of intracellular actin and tubulin cytoskeleton in the proximal tubule was already visible after 15 min of incubation and gradually increased with time, whereas a partial redistribution of actin to the basolateral domain indicated a compromised polarity of the cells. The data also revealed very early (after 15 min) necrotic events in the proximal tubule epithelium, with sloughing of brush border and cell debris into the tubule lumen, detachment of cells from the basal membrane, and opening and widening of the tubule lumen. We conclude that the loss of cellular structure, cytoskeleton, and cell membrane transporters in the nephron epithelium is a very early event in the incubated rat renal cortical slices. PMID- 15895250 TI - A simple way to build a grinder for carbon-fibre electrodes for amperometry or voltammetry. AB - Carbon-fibre electrodes are used widely for studying exocytosis by amperometry. Currently, there are two major methods for insulating fibres so as to leave the tip as the only conductive surface: encapsulation with plastic or glass. The latter offers advantages such as better insulation and a known electro-active surface. In addition, such electrodes are suitable for in vivo electrochemistry because they can penetrate brain tissues. However, the construction of glass encapsulated electrodes requires a grinder to polish the electrode surface with precision. This apparatus is expensive because it needs a very stable motor, a diamond surface and a micromanipulator. We describe the construction of a cheap precision grinder using a computer drive and an old microscope. PMID- 15895251 TI - Essential oil poisoning: N-acetylcysteine for eugenol-induced hepatic failure and analysis of a national database. AB - We present a 15-month-old boy who developed fulminant hepatic failure after ingesting 10 ml of clove oil. After 24 h, the ALT level was in excess of 13,000 U/l, with blood urea and creatinine of 11.8 mmol and 134 micromol/l respectively. The hepatic impairment resolved after intravenous administration of N acetylcysteine so that 6 h later, the ALT level was approximately 10,000 U/l. His liver synthetic function and clinical status improved over the next 4 days. This is the first such case report of its kind in Europe. Analysis of a national database revealed a 14-fold increase in home accidents related to aromatherapy from 1994-1999. Clove oil has important hepatotoxic effects. CONCLUSION: Recent growth in aromatherapy sales has been accompanied by an unfortunate increase in accidental poisoning from these products. Clove oil warrants special attention. Ingesting as little as 10 ml causes hepatotoxicity which can be treated with N acetylcysteine. PMID- 15895252 TI - Reduction of drug leakage by negative-balance isolated pelvic perfusion: correlation between leakage and in-out flow rate in a pig model. AB - PURPOSE: Isolated pelvic perfusion (IPP) therapy exposes target tissues to high doses of anticancer drugs with low systemic concentrations, but the major drawback is drug leakage into the systemic circulation, which often thwarts the increased drug concentration. In this study, the efficacy of altering the in-out flow rate during IPP in order to decrease the leakage was assessed in adult pigs. METHODS: The abdominal aorta and the infrarenal vena cava were occluded with two balloon catheters, blood in the extracorporeal circuit was circulated with twin rotary pumps, and the IPP was performed with platinum. Three sets of in-out flow rates were used, and the degree of drug leakage into the systemic circulation was evaluated. The volume of blood withdrawn was equal to the volume returned (300 ml/min; group A), 5% higher (group B), or 10% higher (group C). The platinum concentrations in the pelvic circulation, systemic circulation, and urine were measured and compared. RESULTS: The average and maximum plasma platinum concentrations in the pelvic circulation did not significantly differ among the three groups. The plasma platinum concentrations in the systemic venous circulation of the three groups significantly (P<0.01) decreased as the volume withdrawn during IPP increased. The percentage of platinum eliminated in the urine during IPP was significantly (P<0.01) lower in group B and C than in group A. CONCLUSIONS: Setting the volume withdrawn higher than the volume returned decreased leakage into the systemic circulation under isolated pelvic perfusion. PMID- 15895253 TI - Isolation of a heat-resistant allergen from the fish parasite Anisakis simplex. AB - The thermal stability of allergenic peptides from the fish parasite Anisakis simplex has not been fully elucidated. This is of special relevance for physicians who should clearly indicate if sensitized patients should avoid ingestion of raw fish only or whether well-cooked fish should also be avoided, if allergenic peptides derived from the parasite remain immunologically detectable. An allergen was purified after heating a crude parasite extract for 30 min. The allergen was further purified by an ethanol fractionation procedure followed by a reversed-phase HPLC. The N-terminal amino acid sequence was obtained. This allergen was detected by 27% of sensitized subjects. The N-terminal amino acid sequence of the 9 kDa allergen showed no similarities to other known proteins. A minor low molecular weight allergen from A. simplex is highly resistant to heating and it could therefore have significant clinical relevance. PMID- 15895254 TI - Rapid diagnosis of Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD) in salmonid fish by a novel DNA amplification method, loop-mediated isothermal amplification (LAMP). AB - A new molecular diagnostic assay was developed for detection of Tetracapsuloides bryosalmonae the causative agent of proliferative kidney disease (PKD) in salmonid fish using a loop-mediated isothermal amplification method (LAMP). The PKD-LAMP assay amplifies the T. bryosalmonae DNA extracted from infected kidney, under constant temperature of 65 degrees C within 1 h. The required equipment for DNA amplification is only a water bath. The amplification products were detected visually by using SYBR green I dye, which turns green in the presence of amplified products and remains orange in its absence, and by electrophoresis without any difference in the sensitivity of both methods. The developed PKD-LAMP assay demonstrated an exceptionally higher sensitivity than the conventional PCR. PKD-LAMP assay was found to be 100-fold more sensitive than the PCR assay. The developed assay is simple, rapid, cost-effective, specific and highly sensitive. The assay is also characterized by its field applicability, as it does not require the use of sophisticated equipment or skilled personnel. PMID- 15895255 TI - Multi-membrane-bound structures of Apicomplexa: I. the architecture of the Toxoplasma gondii apicoplast. AB - Apicomplexan parasites carry a plastid-like organelle termed apicoplast. The previous documentation of four membranes bordering the Toxoplasma gondii apicoplast suggested a secondary endosymbiotic ancestry of this organelle. However, a four-membraned apicoplast wall could not be confirmed for all Apicomplexa including the malarial agents. The latter reportedly possesses a mostly tri-laminar plastid wall but also displays two multi-laminar wall partitions. Since these sectors apparently evolved from regional wall membrane infoldings, the malarial plastid could have lost one secondary wall membrane in the course of evolution. Such wall construction was however not unambiguously resolved. To examine whether the wall of the T. gondii apicoplast is comparably complex, serial ultra-thin sections of tachyzoites were analyzed. This investigation revealed a single pocket-like invagination within a four-laminar wall segment but also disclosed that four individual membranes do not surround the entire T. gondii apicoplast. Instead, this organelle possesses an extensive sector that is bordered by two membranes. Such heterogeneous wall construction could be explained if the inner two membranes of a formerly four-membraned endosymbiont are partially lost. However, our findings are more consistent with an essentially dual-membraned organelle that creates four-laminar wall sectors by expansive infoldings of its interior border. Given this architecture, the T. gondii apicoplast depicts a residual primary plastid not a secondary one as presently proposed. PMID- 15895256 TI - The ratio of maternal to paternal UPD associated with recessive diseases. PMID- 15895257 TI - Functional evaluation of Dent's disease-causing mutations: implications for ClC-5 channel trafficking and internalization. AB - ClC-5 is a member of the ClC family of voltage-gated chloride channels. Loss-of function mutations of its corresponding gene (CLCN5) cause Dent's disease, an X linked kidney disorder, characterized by low-molecular weight proteinuria, hypercalciuria, nephrocalcinosis/nephrolithiasis, and progressive renal failure. Here, we examined the effect of different mutations on function and cellular trafficking of the recombinant protein. Mutant CLCN5 cDNAs were generated by site directed mutagenesis for two premature stop codon variants (R347X and M517IfsX528), and several missense mutations (C221R, L324R, G462 V, and R516 W). We also tested L521R (instead of L521RfsX526 observed) and mutants G506E and R648X (previously reported by others). After heterologous expression in Xenopus oocytes, ClC-5 channel activity and surface expression were determined by two electrode voltage-clamp analysis and ClC-5 surface ELISA, respectively. Except for the R516 W and R648X variants, none of the mutated proteins induced functional chloride currents or reached the plasma membrane. This is readily understandable for the truncation mutations. Yet, the tested missense mutations are distributed over different transmembrane regions, implying that correct channel structure and orientation in the membrane is not only a prerequisite for proper ClC-5 function but also for Golgi exit. Interestingly, the R648X mutant although functionally compromised, displayed a significant increase in surface expression. This finding might be explained by the deletion of a ClC-5 carboxy terminal PY-like internalization signal, which in turn impairs channel removal from the membrane. Our observations further imply that recruitment of ClC-5 to alternative routes (plasma membrane or early endosomes) in the trans-Golgi network is mediated via different signal sequences. PMID- 15895258 TI - Decreased transcription of the human FCGR2B gene mediated by the -343 G/C promoter polymorphism and association with systemic lupus erythematosus. AB - The role for inhibitory Fc gamma receptors class IIb (FcgammaRIIb) in the onset, progression and severity of several animal models of autoimmune diseases is well established. By contrast, the pathogenic potential of FcgammaRIIb in human autoimmune diseases remains largely unknown. Here we report the identification of a polymorphism in the human FCGR2B promoter (dbSNP no. rs3219018) that is associated in homozygosity with systemic lupus erythematosus (SLE) phenotype in European-Americans (OR=11.1, P=0.003). Experimental evidence correlates the polymorphism (a G-C substitution at position -343 relative to the start of transcription) with altered FcgammaRIIb expression and function. The G-C substitution correlated with decreased transcription of the FCGR2B promoter, and resulted in decreased binding of the AP1 transcription complex to the mutant promoter sequence. The surface expression of FcgammaRIIb receptors was significantly reduced in activated B cells from (-343 C/C) SLE patients. These findings suggest that genetic defects may lead to deregulated expression of the FCGR2B gene in -343 C/C homozygous subjects, and may play a role in the pathogenesis of human SLE. PMID- 15895259 TI - An investigation into the use of MR imaging to determine the functional cross sectional area of lumbar paraspinal muscles. AB - The purpose of this study was to investigate the use of magnetic resonance (MR) imaging and image processing software to determine the functional cross-sectional area (FCSA) (the area of muscle isolated from fat) of the lumbar paraspinal muscles. The measurement of the morphology of the lumbar paraspinal muscles has become the focus of several recent investigations into the aetiology of low back pain. However, the reliability and validity of determining the FCSA of the lumbar paraspinal muscles using MR imaging are yet to be reported. T2 axial MR scans at the L1-S1 spinal levels of six subjects were obtained using identical MR systems and scanning parameters. Lean paraspinal muscle, vertebral body bone and intermuscular fat were manually segmented using image analysis software to assign a grey scale range to the MR signal intensity emitted by each tissue type. The resultant grey scale range for muscle was used to determine FCSA measurements for each of the paraspinal muscles, psoas, quadratus lumborum, erector spinae and lumbar multifidus on each scan slice. As various biological, instrument and measurement factors can affect MR signal intensity, a sensitivity analysis was conducted to determine the error associated in calculating FCSA for paraspinal muscle using a discrete grey scale range. Cross-sectional area and FCSA measurements were repeated three times and reliability indices for the FCSA measurements were obtained, showing excellent reliability, intra class correlation coefficient (mean=0.97, range 0.90-0.99) and %SEM (mean=2.6%, range 0.7-4.8%). In addition, the error associated with miscalculation of the grey scale range for the MR signal intensity of muscle was calculated and found to be low with an error of 20 grey scale units at the upper end of the muscle's grey scale range resulting in a very small error in the measured muscle FCSA. The method presented in this paper has a variety of practical applications in areas such as evidence-based rehabilitation, biomechanical modelling and the determination of segmental inertial parameters. PMID- 15895260 TI - Chondroid chordoma of the L5 spinous process and lamina: a case report. AB - Chordoma is a rare bone tumor that originates from the remnants of the notochord. These tumors have axial distribution particularly at the upper and lower ends of the vertebral column. This paper reports a rare occurrence of a chordoma in the posterior elements of the L5 vertebra. A differential diagnosis of a benign tumor (giant cell tumor, aneurysmal bone cyst or osteoblastoma) was made initially. Other differential diagnoses included plasmacytoma and metastasis. The tumor was removed enbloc. Histopathological examination revealed the tumor mass to be chordoma. There were no clinical or radiological signs of recurrence at 21 months follow-up. Chordomas are tumors of the axial skeleton. However, they may occur in unusual sites in ectopic notochordal tissue. The case is being presented for its unusual site of occurrence. PMID- 15895261 TI - A nonverbal test of knowledge attribution: a comparative study on dogs and children. AB - The sensitivity of eleven pet dogs and eleven 2.5-year-old children to others' past perceptual access was tested for object-specificity in a playful, nonverbal task in which a human Helper's knowledge state regarding the whereabouts of a hidden toy and a stick (a tool necessary for getting the out-of-reach toy) was systematically manipulated. In the four experimental conditions the Helper either participated or was absent during hiding of the toy and the stick and therefore she knew the place(s) of (1) both the toy and the stick, (2) only the toy, (3) only the stick or (4) neither of them. The subjects observed the hiding processes, but they could not reach the objects, so they had to involve the Helper to retrieve the toy. The dogs were more inclined to signal the place of the toy in each condition and indicated the location of the stick only sporadically. However the children signalled both the location of the toy and that of the stick in those situations when the Helper had similar knowledge regarding the whereabouts of them (i.e. knew or ignored both of them), and in those conditions in which the Helper was ignorant of the whereabouts of only one object the children indicated the place of this object more often than that of the known one. At the same time however, both dogs and children signalled the place of the toy more frequently if the Helper had been absent during toy-hiding compared to those conditions when she had participated in the hiding. Although this behaviour appears to correspond with the Helper's knowledge state, even the subtle distinction made by the children can be interpreted without a casual understanding of knowledge-formation in others. PMID- 15895262 TI - ISOFIT: a model-based method to measure muscle-tendon properties simultaneously. AB - Estimation of muscle parameters specifying force-length and force-velocity behavior requires in general a large number of sophisticated experiments often including a combination of isometric, isokinetic, isotonic, and quick-release experiments. This study validates a simpler method (ISOFIT) to determine muscle properties by fitting a Hill-type muscle model to a set of isovelocity data. Muscle properties resulting from the ISOFIT method agreed well with muscle properties determined separately in in vitro measurements using frog semitendinosus muscles. The force-length curve was described well by the results of the model. The force-velocity curve resulting from the model coincided with the experimentally determined curve above approximately 20% of maximum isometric force (correlation coefficient R>0.99). At lower forces and thus higher velocities the predicted curve underestimated velocity. The stiffness of the series elastic component determined with direct experiments was approximately 10% lower than that determined by the ISOFIT method. Use of the ISOFIT method can decrease experimental time up to 80% and reduce potential changes in muscle parameters due to fatigue. PMID- 15895263 TI - Cultivation conditions and properties of extracellular crude lipase from the psychrotrophic fungus Penicillium chrysogenum 9'. AB - Among 97 fungal strains isolated from soil collected in the arctic tundra (Spitsbergen), Penicillium chrysogenum 9' was found to be the best lipase producer. The maximum lipase activity was 68 units mL(-1) culture medium on the fifth day of incubation at pH 6.0 and 20 degrees C. Therefore, P. chrysogenum 9' was classified as a psychrotrophic microorganism. The non-specific extracellular lipase showed a maximum activity at 30 degrees C and pH 5.0 for natural oils or at pH 7.0 for synthetic substrates. Tributyrin was found to be the best substrate for lipase, among those tested. The Km and Vmax were calculated to be 2.33 mM and 22.1 units mL(-1), respectively, with tributyrin as substrate. The enzyme was inhibited more by EDTA than by phenylmethylsulfonyl fluoride and was reactivated by Ca2+. The P. chrysogenum 9' lipase was very stable in the presence of hexane and 1,4-dioxane at a concentration of 50%, whereas it was unstable in presence of xylene. PMID- 15895264 TI - Genomic insights into the iron uptake mechanisms of the biomining microorganism Acidithiobacillus ferrooxidans. AB - Commercial bioleaching of copper and the biooxidation of gold is a cost-effective and environmentally friendly process for metal recovery. A partial genome sequence of the acidophilic, bioleaching bacterium Acidithiobacillus ferrooxidans is available from two public sources. This information has been used to build preliminary models that describe how this microorganism confronts unusually high iron loads in the extremely acidic conditions (pH 2) found in natural environments and in bioleaching operations. A. ferrooxidans contains candidate genes for iron uptake, sensing, storage, and regulation of iron homeostasis. Predicted proteins exhibit significant amino acid similarity with known proteins from neutrophilic organisms, including conservation of functional motifs, permitting their identification by bioinformatics tools and allowing the recognition of common themes in iron transport across distantly related species. However, significant differences in amino acid sequence were detected in pertinent domains that suggest ways in which the periplasmic and outer membrane proteins of A. ferrooxidans maintain structural integrity and relevant protein protein contacts at low pH. Unexpectedly, the microorganism also contains candidate genes, organized in operon-like structures that potentially encode at least 11 siderophore systems for the uptake of Fe(III), although it does not exhibit genes that could encode the biosynthesis of the siderophores themselves. The presence of multiple Fe(III) uptake systems suggests that A. ferrooxidans can inhabit aerobic environments where iron is scarce and where siderophore producers are present. It may also help to explain why it cannot tolerate high Fe(III) concentrations in bioleaching operations where it is out-competed by Leptospirillum species. PMID- 15895265 TI - Microbial metabolomics: replacing trial-and-error by the unbiased selection and ranking of targets. AB - Microbial production strains are currently improved using a combination of random and targeted approaches. In the case of a targeted approach, potential bottlenecks, feed-back inhibition, and side-routes are removed, and other processes of interest are targeted by overexpressing or knocking-out the gene(s) of interest. To date, the selection of these targets has been based at its best on expert knowledge, but to a large extent also on 'educated guesses' and 'gut feeling'. Therefore, time and thus money is wasted on targets that later prove to be irrelevant or only result in a very minor improvement. Moreover, in current approaches, biological processes that are not known to be involved in the formation of a specific product are overlooked and it is impossible to rank the relative importance of the different targets postulated. Metabolomics, a technology that involves the non-targeted, holistic analysis of the changes in the complete set of metabolites in the cell in response to environmental or cellular changes, in combination with multivariate data analysis (MVDA) tools like principal component discriminant analysis and partial least squares, allow the replacement of current empirical approaches by a scientific approach towards the selection and ranking of targets. In this review, we describe the technological challenges in setting up the novel metabolomics technology and the principle of MVDA algorithms in analyzing biomolecular data sets. In addition to strain improvement, the combined metabolomics and MVDA approach can also be applied to growth medium optimization, predicting the effect of quality differences of different batches of complex media on productivity, the identification of bioactives in complex mixtures, the characterization of mutant strains, the exploration of the production potential of strains, the assignment of functions to orphan genes, the identification of metabolite-dependent regulatory interactions, and many more microbiological issues. PMID- 15895266 TI - Repeated-batch production of kojic acid in a cell-retention fermenter using Aspergillus oryzae M3B9. AB - A cell-retention fermenter was used for the pilot-scale production of kojic acid using an improved strain of Aspergillus oryzae in repeated-batch fermentations. Among the various carbon and nitrogen sources used, sucrose and yeast extract promoted pellet morphology of fungi and higher kojic acid production. Repeated batch culture using a medium replacement ratio of 75% gave a productivity of 5.3 gL(-1)day(-1) after 11.5 days of cultivation. While batch culture in shake-flasks resulted in a productivity of 5.1 gL(-1)day(-1), a productivity of 5 gL(-1)day( 1) was obtained in a pilot-scale fermenter. By converting the batch culture into repeated batches, the non-productive downtime of cleaning, filling and sterilizing the fermenter between each batch were eliminated, thereby increasing the kojic acid productivity. PMID- 15895267 TI - A preliminary report on patient acceptance of a novel intra-oral lubricating device for the management of radiotherapy-related xerostomia. AB - Management of radiotherapy-related xerostomia is difficult. Saliva substitutes are helpful but the effects are short-lived. The purpose of the study was to develop a prototype intra-oral lubricating device for the management of radiotherapy-related xerostomia and to evaluate patient acceptance. An intra-oral lubricating device was fabricated that incorporated a reservoir in the palatal vault and permitted slow release of saliva substitute by the patient. Preliminary clinical testing was done in five patients with radiotherapy-related xerostomia. A measure incorporating seven questions was used to explore patient acceptance. The device was simple to fabricate using materials available in a technical laboratory. All patients were able to wear the device for at least 4 h per day throughout the test period. The device was considered easy to use and clean. Some impairment of speech and chewing was noted although this appeared to be related to the bulkiness of the reservoir. General oral comfort was improved due to the lubricating effect. The bulk of the reservoir was reduced as a consequence of patient feedback. The design addressed key problems associated with previous lubricating systems. Patient reports on oral functioning with the device in situ provided pivotal information on the device's utility. PMID- 15895268 TI - "Medicamentation" of society, non-diseases and non-medications: a point of view from social pharmacology. AB - This review presents the definition and goals of "social pharmacology", a new branch of clinical pharmacology, investigating relationships between drugs and society through the example of "medicamentation", defined as the use of drugs for social problems previously not requiring drug utilisation (ageing, smoking cessation, vigilance troubles, sleep synchronisation, loss of libido, etc.). The involvement of the different "actors" from our society (patients, physicians, pharmaceutical industries, clinical pharmacologists, regulatory agencies, etc.) in this phenomenon is also discussed. PMID- 15895269 TI - The impact of hospitalisation on the initiation and long-term use of benzodiazepines. AB - BACKGROUND: Inappropriate (long-term) use of benzodiazepines (BZDs) is a reason for concern. Several studies have suggested that hospitalisation may be a determinant for initiation of BZD use as well as for long-term use. However, the available evidence is conflicting. OBJECTIVE: To determine whether hospitalisation induces initiation of BZD use and subsequent long-term use. METHODS: A retrospective follow-up study was conducted. Randomly, 10,000 patients who had been hospitalised were selected (index date). Non-hospitalised patients, matched on age and gender, were sampled from the same living region and assigned the same index date as the corresponding hospitalised patient. Patients were included if adequate medication data were available from 18 months before to 18 months after the index date. Initiation of BZD use was defined as a prescription for a BZD or BZD-related hypnotic without a prescription for any of these drugs during the prior 6 months. Long-term use was defined as a period of consecutive use for at least 6 months following initiation. RESULTS: In this study, 8,681 hospitalised patients and an equal number of non-hospitalised patients were finally included. Overall, the relative risk for initiation of BZD use was almost twice as high [IDR 1.97 (95%CI 1.84-2.10)] among hospitalised patients as in non hospitalised patients. This relative risk was most clearly elevated during the time window from 3 months before to 3 months after hospitalisation [IDR 4.81 (95%CI 4.08-5.67)]. The relative risk for long-term use during the entire 36 month observation period was not higher [IDR 1.04 (95%CI 0.95-1.13)] among hospitalised patients than among non-hospitalised patients. Within the time window of 3 months before and after hospitalisation, the relative risk for long term use was significantly lower for the hospitalised group [RR 0.82 (CI 0.69 0.98)]. CONCLUSION: Our results confirm that hospitalisation is associated with an increased risk for initiation of BZD use; the risk is highest during the 3 months just before and after hospitalisation. However, hospitalisation appeared not to be a determinant for long-term use of BZDs. PMID- 15895270 TI - Press coverage and sales of Xenical in Sweden, 1998-2000. AB - OBJECTIVE: The anti-obesity drug Xenical (orlistat, Roche) was launched on the Swedish market in February 1999. The sales peaked in May 1999 and then declined. The purpose of this study was to investigate the press coverage of Xenical during the period 1998-2000 and, if possible, relate press attention to Xenical sales during this period. METHODS: We analysed all articles published in Sweden's four biggest-selling newspapers mentioning Xenical during the period 1998-2000. Promotion activities aimed at healthcare professionals were measured by registering the number of advertisements in the Journal of the Swedish Medical Association. Sales figures for Xenical were obtained from the National Corporation of Swedish Pharmacies. RESULTS: Approximately twice as many positive effect messages were published than negative or neutral messages. Only six out of 42 positive messages referred to results from scientific studies. The initial high peaks for sales and positive-effect articles coincided with the launch of Xenical on the Swedish market. CONCLUSIONS: It is not possible to discern a causal relationship between press coverage and sales; however, it seems safe to say that the large number of positive articles published before and during the launch greatly increased public awareness of Xenical, thus promoting sales. PMID- 15895271 TI - High-resolution crystal structures of Desulfovibrio vulgaris (Hildenborough) nigerythrin: facile, redox-dependent iron movement, domain interface variability, and peroxidase activity in the rubrerythrins. AB - High-resolution crystal structures of Desulfovibrio vulgaris nigerythrin (DvNgr), a member of the rubrerythrin (Rbr) family, demonstrate an approximately 2-A movement of one iron (Fe1) of the diiron site from a carboxylate to a histidine ligand upon conversion of the mixed-valent ([Fe2(II),Fe1(III)]) to diferrous states, even at cryogenic temperatures. This Glu<-->His ligand "toggling" of one iron, which also occurs in DvRbr, thus, appears to be a characteristic feature of Rbr-type diiron sites. Unique features of DvNgr revealed by these structures include redox-induced flipping of a peptide carbonyl that reversibly forms a hydrogen bond to the histidine ligand to Fe1 of the diiron site, an intra-subunit proximal orientation of the rubredoxin-(Rub)-like and diiron domains, and an electron transfer pathway consisting of six covalent and two hydrogen bonds connecting the Rub-like iron with Fe2 of the diiron site. This pathway can account for DvNgr's relatively rapid peroxidase turnover. The characteristic combination of iron sites together with the redox-dependent iron toggling between protein ligands can account for the selectivity of Rbrs for hydrogen peroxide over dioxygen. PMID- 15895272 TI - Frequency of stages of the seminiferous cycle in the thick-tailed bush baby (Otolemur garnetti), a prosimian primate: possible phylogenetic implications? AB - Spermatogenesis in the thick-tailed bush baby, Otolemur garnetti, was studied using light microscopy. The stages and stage frequencies of the cycle of the seminiferous epithelium were determined using semithin sections stained with methylene blue-azure II. These sections were obtained from the testes of six healthy adult males (n = 6). They revealed 11 stages of the seminiferous epithelial cycle in this species. The mean relative frequencies of the stages I XI were 10.9, 6.0, 5.9, 7.3, 13.2, 10.7, 11.7, 9.2, 7.6, 8.9 and 8.6, respectively. Comparisons were made between the frequency data in the thick tailed bush baby and equivalent data in the rat, hamster, macaque, baboon, chimpanzee and man. There was a significant correlation (P < 0.05) between the Otolemur data and equivalent stage frequency data of two rodent species (rat and hamster) and monkey (Macaca arctoides). However, there was no significant correlation between the present data and those of the baboon, chimpanzee and man. Possible phylogenetic implications of these findings are discussed. PMID- 15895273 TI - [Mycotic infections of the anogenital region]. AB - Mycotic infections of the anogenital region are very common in dermatological practice. While dermatophyte infections are more frequent in men, genital candidosis is a more striking problem in women. The increasing prevalence of non albicans species (esp. C. glabrata, C. krusei, C. guilliermondii) with their resistance against azole derivatives may be responsible for therapeutic failures and a relapsing course in some instances. Most superficial infections of the anogenital area respond satisfactorily to topical antifungal treatment, especially if provocative factors and the possibility of sexual transmission are considered. Systemic treatment is recommended in cases of widespread dermatophyte infections, candidosis or systemic mycosis. PMID- 15895274 TI - An overview of the patient with ataxia. AB - Ataxia, a neurological sign characterized by the incoordination of voluntary movements, is the most prominent manifestation of cerebellar disease. The cardinal features of cerebellar dysfunction involve disturbances of stance, gait, eye movements, muscle tone, skilled movements, and speech. Classification and differential diagnosis of ataxic syndromes have intrinsic complexity owing to the variability in phenotypic presentations and in etiologies, which include trauma, toxic and metabolic causes, neoplasms, immune mechanisms, and genetic diseases. Pure cerebellar symptoms are rarely observed, while the clinical picture of both genetic and sporadic ataxia syndromes is sometimes complicated by the presence of extracerebellar neurological or multisystem extraneural pathology. Clinical presentation and assessment of the patients together with classification, genetic aspects, and principles in differential diagnosis of ataxias are briefly reviewed. PMID- 15895275 TI - Soluble vascular cell adhesion molecule (VCAM) is associated with treatment effects of interferon beta-1b in patients with secondary progressive multiple sclerosis. AB - BACKGROUND: Subcutaneous IFNbeta-1b (Betaferon) is an established immunomodulatory treatment for relapsing remitting MS and active secondary progressive multiple sclerosis (SPMS). It modulates cytokine and adhesion molecule expression but long term in vivo effects of IFNbeta-1b on the immune system are not known in multiple sclerosis. OBJECTIVE: To address the effects of IFNbeta-1b on serum levels for soluble adhesion molecules and cytokine receptors from MS patients. METHODS: Serial blood samples were obtained from 40 patients of the frequent MRI subgroup (20 patients each from the placebo and the IFNbeta-1b treatment group), participating in the European multi-center clinical trial with IFNbeta-1b for secondary progressive MS, at regular intervals for up to 36 months. Soluble adhesion molecules (sVCAM, sICAM-1, sL-Selectin) as well as TNF receptor I and II were analysed in the serum of patients by enzyme linked immunosorbent assays (ELISAs). Monthly brain MRI was performed in 34 of these patients (16 patients from the placebo and 18 from the IFNbeta-1b group) during months 1-6 and 19-24 to monitor disease activity as assessed by newly occurring gadolinium (Gd) enhancing lesions. RESULTS: An early and significant increase in sVCAM and sTNF-RII serum levels was detected in 16 out of 20 patients (80 %) treated with subcutaneous IFNbeta-1b already at month 1 but was absent in all but one patient during placebo treatment (p < 0.01). Raised sVCAM and TNF RII serum levels during months 1-6 inversely correlated with less MRI activity in the 19-24 months treatment interval in the IFNa-1b treatment group ( p = 0.0093 for TNF RII; p = 0.047 for VCAM). CONCLUSIONS: sVCAM and sTNF RII levels in the serum of SPMS patients are increased during IFNbeta-1b therapy and may at least in part explain some of the treatment effects, like reduced immune cell transmigration. PMID- 15895276 TI - Paul Julius Mobius (1853-1907). PMID- 15895280 TI - Severe bone deformities in young children from vitamin D deficiency and fluorosis in Bihar-India. AB - A case-control study was undertaken to understand the etiopathology of the bone deformities among young children in a fluoride-affected village of the Bihar State. Two villages were selected: one village with high fluoride in drinking water (7.9 +/- 4.15 ppm), and the other village with normal levels of fluoride (0.6 +/- 0.31 ppm) as the control village. The source of drinking water was bore wells in both the villages. Two hundred and forty subjects from 54 households (HHs) of the high-fluoride village (HFV) and 1443 subjects from 197 HHs of the control village were selected for the study. Dental mottling (DM) was observed in 50% and skeletal deformities of various forms were observed in 20% of the total population of HFV, whereas, in the control village, DM was 6% and skeletal deformities were absent. The prevalence of both, DM and skeletal deformities was high in the younger age group of 1.5 to 14 years. Genu valgum, genu varum, bowing of tibia, saber shin, and widening of the lower ends of long bones at the wrist were the typical skeletal deformities observed among affected children in the HFV. X-rays of the children with deformities revealed varying degrees of bending of bones and enlargement of epiphyseal ends of metaphyses with fraying of bone and ligamental calcification. A survey indicated significantly low calcium and high phosphorus intake among the population of the HFV as compared to that of the control village, possibly resulting from low intake of milk and high intake of potatoes, respectively. The mean urinary fluoride level was significantly higher in the children of the HPV, both with and without deformities, as compared to that of the control village. The mean serum 25 OHD3 (25 Hydroxy Vitamin D) and calcium levels were significantly lower and alkaline phosphatase activity was significantly higher among the children with deformities as compared to those without deformities from the HFV and the control village. Serum intact parathyroid hormone (IPTH) levels were high in children both with and without deformities in the HFV as compared to those in the control village. No significant differences were observed in the concentration of serum and urinary creatinine, and Cu, and Mg levels between the HFV and the control village. It can be concluded that some of the children from the HFV manifested severe bone deformities (rickets), which were confirmed by the existence of low serum calcium and vitamin D levels. PMID- 15895281 TI - Association of COLIA1 Sp1 alleles with defective bone nodule formation in vitro and abnormal bone mineralization in vivo. AB - Previous work identified a G/T polymorphism affecting a Sp1 binding site in a regulatory region of the COLIA1 gene that predisposes to osteoporotic fractures by affecting bone strength through mechanisms that are partly independent of differences in bone mineral density (BMD). To clarify the mechanisms by which COLIA1 Sp1 alleles influence bone strength we used quantitative backscattered electron imaging (qBEI) to characterize bone mineralization in biopsy samples from subjects of different COLIA1 genotype and studied the ability of osteoblast like cells cultured from subjects of different genotypes to form mineralized bone nodules. The qBEI analysis showed a significant (P = 0.014) reduction in mineralization in bone biopsies from G/T heterozygotes (n = 6) compared with G/G homozygotes (n = 7) and a significant increase in heterogeneity of mineralization (P = 0.017). The in vitro studies showed that osteoblasts derived from G/T heterozygotes (n = 5) were significantly less able to produce mineralized bone nodules than G/G homozygotes (n = 10) at all time-points examined (P < 0.0001). We conclude that carriage of the COLIA1 Sp1 "T" allele is associated with an impaired ability of osteoblast-like cells to form mineralized bone nodules in vitro and with abnormalities of bone mineralization in vivo. This suggests that the increased bone fragility in carriers of the COLIA1 Sp1 allele may result in part from defects in bone mineralization. PMID- 15895282 TI - Inhibition of inflammatory and bone-resorption-inhibitory effects of alendronate by etidronate. AB - Among the bisphosphonates (BPs), the aminobisphosphonates (aminoBPs) have much stronger bone-resorption-inhibitory activities (BRIAs) than nonaminobisphosphonates (nonaminoBPs). However, aminoBPs have inflammatory effects. We previously reported that in mice: (i) all aminoBPs tested (10-40 micromol/kg) induced various inflammatory reactions (including induction of histidine decarboxylase), whereas clodronate (a non-aminoBP) (10-160 micromol/kg) inhibited these reactions; and (ii) a clear sclerotic line (tentatively called the BP line) was detectable in the tibia by radiography a few weeks after a single injection of either alendronate (a typical aminoBP) (1.6 micromol/kg) or clodronate (160 micromol/kg), and this BP-line formation (a marker for the BRIAs of BPs) was not reduced in mice given both alendronate and clodronate. In this study, using this murine model, we compared clodronate, etidronate (another typical non-aminoBP), alendronate, etidronate + alendronate, and clodronate + alendronate in terms of their inflammatory effects and/or BP-line formation. For BP-line formation, 480 micromol/kg etidronate was needed (single injection). At 160 micromol/kg, etidronate inhibited the histidine decarboxylase induction, but not the other inflammatory reactions induced by alendronate. However, etidronate (unlike clodronate) also inhibited alendronate-induced BP-line formation (even at 40 micromol/kg). Etidronate (160 micromol/kg) also inhibited the physicochemical changes in the tibia induced by six, weekly injections of alendronate. Therefore, depending on the dose, etidronate can inhibit alendronate's inflammatory actions and its BRIA. These results, together with those reported previously, suggest that a strategy utilizing clodronate (but not etidronate) plus an aminoBP might prevent or reduce the inflammatory side effects induced by aminoBPs while preserving their powerful BRIAs. We discuss the mechanisms underlying the antagonism between aminoBPs and non-aminoBPs. PMID- 15895283 TI - Do young New Zealand Pacific Island and European children differ in bone size or bone mineral? AB - Although Pacific Island adults have been shown to have larger bones and greater bone mineral density than caucasians, no previous studies have been undertaken to determine whether differences are present in prepubertal children. Forty-one Pacific Island children (both parents of Pacific Island descent) and 38 European children, aged 3 to 7 years, living in New Zealand were studied. Heights and weights were determined by simple anthropometry and body mass index (BMI, kg/m2) was calculated. Body composition, bone size, and bone mineral content (BMC, g) were measured by dual energy X-ray absorptiometry (DXA) of the total body and the non-dominant forearm. Compared to European children, in data adjusted for age and gender, Pacific Island children had significantly greater (P < 0.05) BMC in the total body (12%), the ultradistal radius (16%), and the 33% radius (8%), and also greater total body bone area (10%). Bone mineral density (BMD, g/cm2) was higher only at the ultradistal radius (11%). However, after adjustment for body weight, in particular lean mass, no differences were seen between Pacific Island and European children in any bone measure. The larger bone area and BMC of young Pacific Island children can be explained by their greater height and weight. Therefore, this study has shown that prepubertal Pacific Island children do not have greater bone size or BMC for their weight. PMID- 15895284 TI - Cyclic tensile stretch stimulates the release of reactive oxygen species from osteoblast-like cells. AB - It is known that the excessive generation of reactive oxygen species (ROS) is a significant factor in tissue injury observed in many disease states. To determine whether extreme levels of mechanical stress applied to osteoblasts enhances ROS synthesis, we loaded cyclic tensile stretch on osteoblast-like HT-3 cells. Cyclic tensile stretch loaded on these cells clearly enhanced ROS synthesis in a time- and magnitude-dependent fashion. Cyclic tensile stretch also enhanced superoxide dismutase (SOD) activity. The disruption of microfilaments with cytochalasin D abolished the stress-induced ROS synthesis. Rotenone, an inhibitor of the mitochondrial electron transport chain, enhanced stress-induced ROS synthesis. These data suggest that actin filament and mitochondria are involved in this action. PMID- 15895285 TI - Morphometric changes in the epiphyseal plate of the growing and young adult male rat after long-term salmon calcitonin administration. AB - The function of the epiphyseal plate is related to the differentiation and maturation of the chondrocytes, especially of the hypertrophic zone. Salmon calcitonin exerts a positive effect on chondrocytes of different types of cartilage, e.g., articular cartilage, osteochondral callus formation, and the epiphyseal plate. In the present study, the effect of long-term daily salmon calcitonin treatment upon epiphyseal plate function was examined in 80 male Wistar rats aged 12 weeks at the beginning of the experiment. A daily dose of 6 IU of salmon calcitonin enhanced the number of the chondrocytes of the hypertrophic zone of the upper tibial epiphyseal plate, increased the mean thickness of the epiphyseal plate, and accelerated the longitudinal growth of long bones. It was found that the peripheral growth of the epiphyseal plate was delayed after calcitonin treatment in comparison with the placebo-treated animals. The most effective period for calcitonin treatment on epiphyseal plate function seems to be the late accelerated period of growth, i.e., puberty. In conclusion, long-term salmon calcitonin treatment has a beneficial effect on longitudinal skeletal growth and this effect remains throughout the adult life of the animal. Salmon calcitonin does not enlarge the surface of the epiphyseal plate. PMID- 15895286 TI - Assessment of tailor-made prevention of atherosclerosis with folic acid supplementation: randomized, double-blind, placebo-controlled trials in each MTHFR C677T genotype. AB - This study aimed at assessing the effect of folic acid supplementation quantitatively in each MTHFR C677T genotype and considered the efficiency of tailor-made prevention of atherosclerosis. Study design was genotype-stratified, randomized, double-blind, placebo-controlled trials. The setting was a Japanese company in the chemical industry. Subjects were 203 healthy men after exclusion of those who took folic acid or drugs known to effect folic acid metabolism. Intervention was folic acid 1 mg/day p.o. for 3 months. The primary endpoint was plasma total homocysteine level (tHcy). In all three genotypes, there were significant tHcy decreases. The greatest decrease was in the TT homozygote [6.61 (3.47-9.76) micromol/l] compared with other genotypes [CC: 2.59 (1.81-3.36), CT: 2.64 (2.16-3.13)], and there was a significant trend between the mutated allele number and the decrease. The tHcy were significantly lowered in all the genotypes, but the amount of the decrease differed significantly in each genotype, which was observed at both 1 and 3 months. Using these time-series data, the largest benefit obtained by the TT homozygote was appraised as 2.4 times compared with the CC homozygote. Taking into account the high allele frequency of this SNP, this quantitative assessment should be useful when considering tailor-made prevention of atherosclerosis with folic acid. PMID- 15895287 TI - Highly polymorphic human CYP4A11 gene. AB - The cytochrome P450 (CYP) is a monooxidase, which regulates metabolism of drugs and fatty acids in the liver and kidney. Among isoforms of the CYP4A subfamily, CYP4A11 is a major lauric acid (medium-length fatty acids) omega hydroxylase and is involved in the balance of lipids in the human liver. We performed direct DNA sequencing in 24 unrelated Korean individuals in the whole gene, including the 1 kb upstream region of CYP4A11. Seventy sequence variants were identified: six in exons, including two nonsynonymous SNPs; 60 in introns; and four in 3'UTR. In comparison with SNPs enrolled in the SNP database (dbSNP) of the National Center for Biotechnology Information (NCBI), 26 novel polymorphisms (24 in introns and two in 3'UTR) were identified in Korean subjects (n=24). The distributions of polymorphisms confirmed were significantly different from those in the dbSNP of the NCBI. Information clarified in this study would provide valuable for further studies, including genetic association studies for various diseases and drug responses. PMID- 15895289 TI - Evidence of changes in sural nerve conduction mediated by light emitting diode irradiation. AB - The introduction of light emitting diode (LED) devices as a novel treatment for pain relief in place of low-level laser warrants fundamental research on the effect of LED devices on one of the potential explanatory mechanisms: peripheral neurophysiology in vivo. A randomised controlled study was conducted by measuring antidromic nerve conduction on the peripheral sural nerve of healthy subjects (n=64). One baseline measurement and five post-irradiation recordings (2-min interval each) were performed of the nerve conduction velocity (NCV) and negative peak latency (NPL). Interventional set-up was identical for all subjects, but the experimental group (=32) received an irradiation (2 min at a continuous power output of 160 mW, resulting in a radiant exposure of 1.07 J/cm(2)) with an infrared LED device (BIO-DIO preprototype; MDB-Laser, Belgium), while the placebo group was treated by sham irradiation. Statistical analysis (general regression nodel for repeated measures) of NCV and NPL difference scores, revealed a significant interactive effect for both NCV (P=0.003) and NPL (P=0.006). Further post hoc LSD analysis showed a time-related statistical significant decreased NCV and an increased NPL in the experimental group and a statistical significant difference between placebo and experimental group at various points of time. Based on these results, it can be concluded that LED irradiation, applied to intact skin at the described irradiation parameters, produces an immediate and localized effect upon conduction characteristics in underlying nerves. Therefore, the outcome of this in vivo experiment yields a potential explanation for pain relief induced by LED. PMID- 15895290 TI - Arteriovenous malformation of the floor of the mouth: a case report. AB - Arteriovenous malformations of the head and neck are rare lesions with unclear pathogenesis. They usually present during childhood, growing proportionately to the child. Although preoperative superselective embolization followed by surgical resection is the treatment of choice, complete removal is often not feasible, leading to high recurrence rates. The case of a patient with an arteriovenous malformation of the floor of the mouth diagnosed late in her adulthood and its management are presented. PMID- 15895291 TI - Audiometric evaluation of carriers of the connexin 26 mutation 35delG. AB - Mutation in a gap junction protein gene (GJB2 also named connexin 26) is a major cause of autosomal recessive congenital deafness, which is responsible for about 80% of the cases in Mediterranean families, but actually little is known about the influence of GJB2 mutations on the hearing of obligate carriers. We examined GJB2 35delG mutation carrier individuals to test the possible presence and incidence of audiometric abnormalities among carriers of 35delG mutations. Tonal audiometric analysis was performed on a 35delG mutation carrier group (H) and on a non-carrier control group (N). Audiometric evaluations in the control group showed the presence of thresholds within normal limits at all frequencies, while carriers of 35delG mutations presented a decrease of hearing principally at 6,000 and 8,000 Hz. The difference at 6,000 and 8,000 Hz between groups H and N is statistically significant. PMID- 15895292 TI - Malignant cervical teratoma: report of a case in a newborn. AB - Malignant cervical teratoma (MCT) usually appears in newborns as an enlarging mass of the neck that causes respiratory distress, requiring prompt airway control. We report a case of MCT in an infant electively delivered at 32 weeks to prevent airway impairment. At first, the preoperative diagnosis was hygroma of the neck, and a surgical excision was performed when the newborn was 9 days old. Diagnosis was benign extragonadic immature teratoma, but it was changed in MCT when cervical metastases appeared and the alpha-fetoprotein (AFP) level increased. Subsequent surgical procedures and chemotherapy were necessary. The child has been free from disease and healthy for 7 years since the last surgery. The preoperative diagnosis of MCT is difficult because of its rarity and non specific clinical findings. Surgical excision is required for an adequate cure and airway repair; a long-term follow-up is mandatory to promptly treat any recurrence. PMID- 15895293 TI - Partial release of the superficial medial collateral ligament for open-wedge high tibial osteotomy. A human cadaver study evaluating medial joint opening by stress radiography. AB - To perform an open-wedge high-tibial osteotomy (HTO), the medial proximal tibia is frequently exposed by partial distal release of the overlying insertion of the medial collateral ligament (MCL). Biomechanically, any release of the MCL can increase knee laxity when valgus stress is applied. Clinically however, post surgical valgus instability following HTO with partial MCL release is an uncommon complication. It is known that the open-wedge procedure can re-tention an intact MCL by the width of the base of the wedge. However, this re-tentioning effect is uncertain in small wedge sizes, preexisting medial compartment laxity and in the presence of a partially detached MCL. Considering the good clinical results after HTO, we hypothesized that a partial release of the superficial MCL for HTO does not play a crucial role in stabilizing valgus forces in the human knee. We therefore measured the effect of partial versus complete release of the superficial MCL to determine medial knee laxity represented by the amount of medial joint opening (MJO) under valgus stress in this human cadaver study. In ten knee pairs, the superficial and deep MCL were sectioned in sequence with a standardized abduction force of 15 kp with a Scheuba apparatus applied. In group 1 (5 knee pairs), the superficial MCL was completely sectioned whereas in group 2 (5 knee pairs), sectioning of the superficial MCL was restricted to the anterior border to mimic the surgical exposure for an HTO. To account for the interindividual variability of ligamentous laxity, only increments of MJO within knee pairs were statistically evaluated. Stress radiography did not reveal any significant differences in increments of MJO between knee pair specimens with complete versus partial release of the superficial MCL. We disproved our hypothesis and concluded that the anterior fibers of the superficial MCL do play a crucial role in maintaining valgus stability in this biomechanical setting. Therefore, the release of the superficial MCL for open-wedge HTO should be kept to a minimum to decrease the potential of late valgus instability. This is especially important in patients with small wedge sizes and medial compartment laxity since the anterior MCL fibers are the main contributor to medial joint stability and the re-tentioning effect of the remaining MCL fibers is presumably decreased. PMID- 15895294 TI - The Oxford unicompartmental knee prosthesis: an independent 10-year survival analysis. AB - One hundred forty-nine medial prostheses were implanted in 140 patients between 1988 and 1996. After a mean of 67 months 28 patients had died, without the need for revision. Seventeen prostheses were lost to follow-up. Revision surgery using a total knee prosthesis was performed in 16 cases. In four others, a lateral prosthesis was implanted subsequently to a medial one. One of these four was revised to a total knee prosthesis 6 years later. In another four cases, late complications of the meniscal bearing were treated with replacement of this bearing. The surviving prostheses were seen back after a mean of 126 months. The cumulative survival rate at 10 years was 82% for the whole population and 84% when knees with a previous high tibial osteotomy were excluded. Since these results compare poorly to the survival of total knee arthroplasty, this prosthesis is not the first-choice implant. Because it preserves a maximum of bone stock and is revised to a total prosthesis almost without difficulty, it is the first-choice implant for medial unicompartmental osteoarthritis in patients younger than 65. Further research is mandatory to confirm that this prosthesis very rarely needs revision in patients older than 75. It should not be used in osteotomized knees. PMID- 15895295 TI - Proteinase 3 hydrolysis of peptides derived from human elastin exon 24. AB - In normal and pathological tissues, elastin-derived peptides proceed of elastin degradation by polymorphonuclear leukocyte proteases: elastase, cathepsin G and proteinase 3. They were demonstrated to have a chemotactic activity, to promote cell proliferation and protease release, . . .. To be biologically active, their structures, which reflect elastase specificity, must adopt a beta-turn conformation which accommodate to the cell surface-located elastin binding protein. In this study, we establish that human elastin exon 24-derived peptides containing at least two repeated VGVAPG sequences are hydrolyzed by the proteinase 3 (Pr3). As shown by mass spectrometry analyses, the demonstrated cleavage sites are in agreement with previously reported Pr3 substrate specificity and its lengthy substrate binding site. The characterization of the Pr3-generated products indicate that they contain at least one GXXPG sequence known to stimulate cellular effects after binding to the elastin receptor. PMID- 15895296 TI - Recurrent laryngeal nerve damage and phonetic modifications after total thyroidectomy: surgical malpractice only or predictable sequence? AB - Modifications of phonation occurring after total thyroidectomy (TT) are usually attributed to surgical malpractice, but other causes of voice impairment even in nonoperated subjects should also be taken into account. This study analyzes 208 patients who underwent TT from January 1, 1999 through December 31, 2001. Follow up ended on December 31, 2003. Only cases in which the surgeon ruled out the possibility of operative damage to the laryngeal nerves were included. All patients underwent pre- and postoperative clinical and instrumental nose and throat examination (NTE). Preoperatively, 86 patients (41%) showed hoarseness or dysphagia: 4 (2%) monoplegia and 12 (6%) hypomobility of the vocal cords due to impaired function of the recurrent laryngeal nerve (RLN); 6 (3%) cord hypotonia due to impairment of the superior laryngeal nerve (SLN); 34 (16%) dysphagia: and 30 (14%) hoarseness due to other causes. At follow-up 1 month after surgery, 71 patients (34%) had an onset of previously absent signs and symptoms: 8 (4%) had palsy of one vocal cord (2% permanent); 6 (3%) had cord hypomobility (all temporary); 12 (6%) had cord hypotonia due to disease of the SLN, 4 of which (2%) were permanent; 44 patients (21%) had symptoms due to scarring and adhesions between the laryngotracheal axis and the prethyroid muscles and between these and the skin. One patient (0.5%) had a nodular cord lesion that occurred after 3 months. Overall, more than one-third of the patients had preoperative voice modifications or swallowing impairment, around one-third had these problems after TT, and less than one-third were free of pre- and postoperative complications. The surgeon's care to avoid damage to the anatomica integrity of the of laryngeal nerves does not exclude functional problems of the nerves and of laryngeal dynamics. In fact, such problems could be referred to outcomes linked to the operation itself (hematoma, edema, scarring adhesion) or to events that only temporarily follow surgery but must be considered as an unavoidable sequel (e.g., neuritis, viral neuritis, myopathy). The patient should undergo a careful clinical and instrumental NTE to detect conditions prior to surgery, and the information provided by the surgeons should be thorough to allow the patient to be aware of all possible sequels and consequences. PMID- 15895297 TI - Inhibition of gastric cancer cell adhesion in nude mice by inraperitoneal phospholipids. AB - Adhesion of tumor cells to mesothelial cells or extracellular matrix components is a pivotal step in developing peritoneal dissemination after gastric cancer. As phospholipids were found to reduce adhesion formation, especially at sites of peritoneal lesions, we assessed the inhibition of attachment of NUGC-4 gastric cancer cells by local treatment with phospholipids to the peritoneum in nude mice. Gastric cancer cells (1xl0(6)) suspended in either normal saline (controls) or phospholipid suspension 75 mg/kg body weight (PL75) or 150 mg/kg (PL150) were injected intraperitoneally into 90 female BALB/c nu/nu mice. The treatment groups were subdivided into animals with defined peritoneal lesions and animals without lesions. After 30 days the extent of peritoneal carcinosis and the Peritoneal Cancer Index were evaluated. Statistical analysis was performed with two factorial ANOVAs. The level of significance was adjusted according to Bonferrorni (alpha = 0.00278). During a 90-day observation period the survival rate was determined using the log rank test. After 30 days the intraperitoneal tumor volume was reduced by PL150 up to 0.6 ml (SEM 0.16) and 0.48 ml (SEM 0.09) in mice with peritoneal lesions compared to 0.9 ml (SEM 0.2) and 0.9 ml (SEM 0.1) in the control group (P = 0.04). The mean area of tumor adhesion amounted to 145 mm(2) (SEM 17) (P = 0.08) and 164 mm(2) (SEM 32.8) (P = 0.049) with peritoneal lesions after treatment with PL150 [controls: 216 mm(2) (SEM 28.5) and 245 mm(2) (SEM 29.3)]. The peritoneal cancer index was 16.4 (SEM 1.7) in the control group and 9 (SEM 1.68) with PL150 (P = 0.0002). In the subgroup with peritoneal lesions, the respective values were as follows: controls: 20.8 (SEM 0.85); PL 150:14.3 (SEM 1.07) (P = 0.0001). We found a prolonged survival rate after treatment with PL150. However, this effect was not significantly different to that seen in the control group. Treatment with PL75 had no significant influence. Phospholipids may be an efficacious and economic tool for reducing peritoneal tumor cell adhesion and consequently the development of peritoneal carcinosis after resection of gastric cancer. PMID- 15895298 TI - Recurrence in pediatric craniopharyngiomas: analysis of clinical and histological features. AB - OBJECTIVE: The purpose of this study was to investigate the recurrence pattern and significance of various clinical and histological features as predictors of recurrence in pediatric craniopharyngiomas. METHODS: A series of 116 pediatric craniopharyngiomas (68 boys and 48 girls; age range, 1.6-18 years) was reviewed. Mean follow-up period was 18.53 months. Tumors recurred in 15 patients within 96 months [mean recurrence-free survival (RFS), 12.67 months]. Of the recurrence cases, 2 had complete (mean RFS, 16 months) and 13 had subtotal tumor excision (mean RFS, 9.03 months). Histologically, an adamantinous pattern was seen in 95% of cases, whereas a papillary pattern was noted in 5%. Brain tissue was included in 41 cases. In 32 of 41 cases, brain invasion was noted, and all were of adamantinous histology. No correlation was noted of histopathological subtyping or brain invasion with recurrence. CONCLUSIONS: The significant clinical factors associated with recurrence included extent of resection, tumor size >4 cm, and cystic tumors. PMID- 15895299 TI - alpha-Synuclein-immunoreactive structure formation is enhanced in sympathetic ganglia of patients with multiple system atrophy. AB - We immunohistochemically examined the sympathetic ganglia (SG) and brains of 26 patients with multiple system atrophy (MSA), 19 age-matched controls, and 25 patients with amyotrophic lateral sclerosis (ALS). alpha-Synuclein-immunoreactive structures were found in the neuronal cytoplasm and processes of the SG in 11 of the 26 MSA cases (42.3%) and 1 of the 25 ALS cases (4%), but not in the 19 controls. No alpha-synuclein-immunoreactive structures were found in Schwann cells or the neuronal nucleus. Mean disease duration of MSA cases with alpha synuclein-immunoreactive structures was significantly longer than that of MSA cases without alpha-synuclein-immunoreactive structures. alpha-Synuclein immunoreactive structures in 4 cases proved to be Lewy bodies (LB) based on hematoxylin-eosin staining. A few LB were also found in the brains of 3 of these 4 cases. In the other 7 MSA cases, diffuse or focal neuronal cytoplasmic aggregates and swollen neurites were detected with alpha-synuclein immunostaining, but not with hematoxylin-eosin staining. However, a few LB-like structures with ring-like staining were observed in those aggregates, which suggested those aggregates had progressed to form LB. Immunoelectron microscopically, those aggregates were composed of filaments and granular materials which closely resembled the ultrastructural features of LB. We inferred that alpha-synuclein aggregates found in the SG in our study evidenced LB-related pathologies. MSA, a type of synucleinopathy, is characterized by glial cytoplasmic inclusions in oligodendrocytes, but also frequently develops LB pathology in the late stage, especially in the SG. PMID- 15895300 TI - Risk factors and early signs of Alzheimer's disease in a family study sample. Risk of AD. AB - OBJECTIVE: Several predictors of Alzheimer's disease (AD) have been identified. However, the relevance and independent contribution of risk factors and of possible early signs such as mild cognitive impairment and subjective memory impairment on the development of AD has not been investigated prospectively in a cohort of non-demented elderly including first-degree relatives of AD subjects. METHOD: The development of AD was investigated in 757 non-demented elderly. Initial diagnoses were made from personal interviews. Information on 633 subjects after 4.7 +/- 1.2 years (mean +/- SD) was obtained either from personal or family history interviews. Using forward logistic regression analysis, predictors were identified by comparing their presence in 38 subjects who developed AD and 577 subjects who remained non-demented. RESULTS: The most important predictors of later Alzheimer's disease were increased age (Odds ratio OR = 1.086/additional year, p < 0.001), initial subjective memory complaints (OR = 2.68, p = 0.019), initial mild cognitive impairment (OR = 2.51, p = 0.032) and female gender (OR = 2.84, p = 0.069). Exploratory analysis revealed that previous depression after the age of 60 years (OR = 2.37, p = 0.033) and the presence of the apolipoprotein E4 allele (OR = 2.49, p = 0.043) individually predicted new AD during follow-up. A positive family history of AD (i. e. being a first degree relative of a subject suffering from AD) did not significantly influence the development of AD (p > 0.2). CONCLUSIONS: Increased age, the presence of mild cognitive impairment, subjective memory impairment and gender are the most relevant independent predictors of later Alzheimer's disease that may be used in combination for clinical prediction of AD. PMID- 15895302 TI - Dimensions of the Typus melancholicus personality type. AB - The Typus melancholicus personality type (TMP) is characterised by orderliness, conscientiousness and interpersonal dependence. Several standardised instruments have been developed for the assessment of the Typus melancholicus personality. To date there has been no systematic comparison of these instruments and in particular it has been unclear whether TMP represents a single trait or a personality trait constellation. The aim of this study was the comparison of four TMP questionnaires and the investigation of the dimensionality of the personality as revealed by these questionnaires. The factorial validity of four TMP questionnaires was examined based on a sample of n = 264 psychiatric inpatients and normal controls. In a factor analysis of the items of the TMP questionnaires, four dimensions could be differentiated: Dependence, Intolerance of Ambiguity, Norm-Orientation, and Perfectionism. Psychometric evaluation showed good values for the individual items and the new TMP scales. The four subscales had a differential correlation profile in relation to the dimensions of the five-factor model of personality. The TMP scales could distinguish a group of depressed patients from a group of normal controls. The results show that TMP personality is not a single trait but consists of four related but separate traits. These can be clearly distinguished from those of the five-factor model of personality. The analysis of the TM concept therefore also represents a theoretical perspective for the integration of the personality characteristics which are relevant for depression. Based on this analysis, we constructed a multidimensional TMP inventory which forms the basis for the investigation of the effect of TM personality on clinical outcome and on psychotherapeutic treatment. PMID- 15895303 TI - Falls in outpatients with Parkinson's disease: frequency, impact and identifying factors. AB - Falls are one of the most serious complications of gait disturbances in patients with Parkinson's disease (PD). Among previous reports, the percentage of patients with PD who fall varies between 38% to 68%. We sought to determine the frequency of falls and the factors associated with falls in a group of patients with idiopathic PD who attended an outpatient, tertiary movement disorders clinic. 350 ambulatory, non-demented patients (230 males) were studied. Mean age was 69.7+/ 10.6 years (range: 43-97 yrs) and mean duration of PD symptoms was 8.6+/- 6.2 years (range: 1-33 yrs). Assessments included characterization of demographics, disease duration, disease severity as measured by the Hoehn and Yahr Scale (H&Y), co-morbidities, the presence of depressive symptoms, the presence of urinary incontinence, use of anti-parkinsonian medications, and two performance-based tests of balance and gait (tandem standing and Timed Up & Go). Fall history was determined during three time periods: previous week, previous month, and previous year. Univariate and multivariate logistic regression models were applied to evaluate the relationship between the above-mentioned factors and falls. 46% of the subjects reported at least one fall in the previous year and 33% reported 2 or more falls and were classified as Fallers. Fallers had significantly more prolonged and advanced PD compared with Non-fallers (p=0.001 and p<0.001, respectively). Urinary incontinence was the factor most closely associated with falls (crude and adjusted OR were 1.95 and 5.89, respectively). Other factors significantly associated with fall status included increased Timed Up & Go times and increased PD duration. These findings confirm that falls are a common problem among patients with advanced PD and suggest easily measurable features that may be used to prospectively identify those PD patients with the greatest risk of falls. PMID- 15895304 TI - Relapse of primary CNS lymphoma after more than 10 years in complete remission. PMID- 15895305 TI - Automatic respiration in amyotrophic lateral sclerosis. PMID- 15895306 TI - Discrimination of white matter lesions and multiple sclerosis plaques by short echo quantitative 1H-magnetic resonance spectroscopy. AB - Multiple sclerosis (MS) plaques and age related white matter lesions (WML) are of similar morphological appearance on T2 weighted MRI. Therefore their differentiation is sometimes crucial. Proton magnetic resonance spectroscopy ((1)H-MRS) adds metabolic information to conventional imaging and may help to distinguish inflammatory MS plaques from vascular related WML. This study was performed to evaluate the metabolite pattern in MS plaques and WML. 15 MS patients, 14 elderly individuals with WML and 16 controls were investigated by conventional MRI and short echo quantitative (1)H-MRS (TE: 30ms, TR: 3000ms). The mean metabolite concentrations in normal control white matter (NCWM), MS plaques and WML were: t-NAA: 8.96 mmol/l (SD: 0.93) vs 6.79 mmol/l (SD: 1.99) vs 7.18 mmol/l (SD: 1.41); Cho:1.66 mmol/l (SD: 0.4) vs 1.49 mmol/l (SD: 0.45) vs 1.46 mmol/l (SD: 0.34); PCr:5.64 mmol/l (SD: 0.83) vs 4.9mmol/l (SD: 1.3) vs 4.95 mmol/l (SD: 0.86); myo-Ins: 4.57 mmol/l (SD:1.05) vs 6.34 mmol/l (SD: 2.03) vs 4.5 mmol/l (SD: 0.96). t-NAA reduction in MS plaques and WML was significant compared with controls (p 0.05). Tritrac activity counts were highly correlated with DLW-PAEE (r = 0.81). High correlations between Digiwalker and DLW were observed when body weight was considered. A low correlation (r = 0.49) was observed between the physical activity energy expenditure by SAPAC and DLW. In conclusion, with some limitations the Tritrac and Digiwalker can provide useful and accurate information about PA and EE in 7- to 10-year-old children. PMID- 15895320 TI - Preferred pedal rate: an index of cycling performance. AB - The purpose of this study was to determine whether preferred pedal rate (PPR) could be used as an index of cycling performance. Thirteen competitive cyclists cycled at PPR during a graded test and a supra-threshold constant power test (CPT). The results showed that PPR values reported in CPT were correlated to both peak power determined from the graded test and exhaustion time assessed in CPT. Furthermore, multiple regression revealed that PPR values and P (peak) were two predictors of exhaustion time in CPT. Hence, this study suggests that coaches and sports scientists have to take into account PPR values complementary to P (peak) in order to evaluate the capacity of cycling performance. PMID- 15895321 TI - Rowing performance and estimated training load. AB - We related the rowing performance and the associated physiological parameters to the training load as estimated by a questionnaire addressing the mean habitual weekly energy expenditure (MHWEE) of twenty-one international and national level oarsmen. The questionnaire also addressed the energy expenditure during training (EET) sessions classified as low- (EE1), moderate- (EE2), and high-intensity (EE3). To evaluate the physiological capability of the oarsmen, they performed incremental exercise to determine their maximal oxygen uptake (V.O(2max)) and the V.O(2) relative to V.O(2max) corresponding to the 4 mmol.l(-1) blood lactate concentration (V.O(2)4 %). The mean work rate sustained during a 2000-m all-out event on a rowing ergometer was considered as the rowing performance. On average, the rowers spent 16.4 +/- 1.0 h.wk(-1) in training with 56 +/- 3 % of the time spent on the water. EET represented 43.5 +/- 1.7 % of MHWEE. Rowing performance and V.O(2max) were both related to MHWEE and EET. Also, rowing performance was related to EE1, EE2, and EE3. In contrast, V.O(2)4 % was not related to the estimated energy expenditures. These results suggest that rowing performance and V.O(2max) are related to training load while V.O(2)4 % was not in the present group of highly trained oarsmen. PMID- 15895322 TI - Reliability of sprint test indices in well-trained cyclists. AB - The study aim was to assess reliability of repeated laboratory sprint tests in well-trained endurance cyclists. Eleven male cyclists (mean +/- standard deviation: 27 +/- 6 yr, 1.79 +/- 0.04 m, 70.1 +/- 3.3 kg) performed a maximal 30 second sprint test on four separate occasions using their own bicycle fitted with an SRM powermeter on a Kingcycle air-braked ergometer. Peak power output (W (peak)), mean power (W (mean)) and an index of fatigue (FI) were calculated. Three minutes post sprint, capillarised blood lactate measurements were taken and analysed. No significant differences (p > 0.05) were found between trials for W (peak), W (mean), FI and blood lactate concentration. Repeatability of W (peak), W (mean), and fatigue index improved across trials 2 and 3 when compared to trials 1 and 2. The highest CV for these variables was recorded between trials 3 and 4. The CV for W (peak) was 4.5 +/- 1.6 %, W (mean) 2.4 +/- 1.2 %, and FI 17.2 +/- 7.1 %. Intraclass reliability coefficients were 0.93 (95 % CI 0.84 - 0.98), 0.94 (95 % CI 0.86 - 0.98) and 0.89 (95 % CI 0.69 - 0.95) respectively. Blood lactate concentration ranged between 5.35 and 14.52 mmol.l(-1), with a mean CV of 12.1 +/- 4.2 %. The CV for trials 2 and 3 revealed the highest CV for blood lactate concentration (15.1 %). The lowest CV for this variable (10.2 %) was recorded between trials 3 and 4. The intraclass reliability coefficient for blood lactate concentration was 0.79 (95 % CI 0.58 - 0.93). The results of this study indicate that there is no improvement in the reliability of sprint test indices when assessing well-trained, experienced cyclists, riding on their own cycle equipment. PMID- 15895323 TI - Biomechanics and energetics of basketball wheelchairs evolution. AB - The aim of this study was to investigate metabolic demand and mechanical work of different basketball wheelchairs that represented significant stages of its evolution from 1960 to date. Four subjects pushed each model on a basketball court at different speeds (from 0.90 to 2.35 m.s(-1)). During the trials, oxygen consumption was measured. Also, the different forms of mechanical work involved in the exercise were investigated. The oxygen consumption decreased from the oldest model to the next ones, remaining then quite constant. This was also the same with breathing and pushing frequencies. Both the work against air drag and rolling resistance decreased, air drag always played a minor role due to the low speeds investigated. The total mechanical work was highest in the oldest wheelchair and lowest in the newest one. The efficiencies were found similar for all the chairs but the most recent one (less efficient). Already by the 1970's the wheelchair economy had reached an acceptable level, at least partially because of its improved ergonomics. Yet, when focusing on the efficiency, the surprisingly low value with the newest model suggests factors other than the economy (need of better balance, responsiveness, and ground grip) as determinants of the evolution of this device. PMID- 15895324 TI - On the methodology of the Conconi test. PMID- 15895326 TI - Strong association of the Y402H variant in complement factor H at 1q32 with susceptibility to age-related macular degeneration. AB - Using a large sample of cases and controls from a single center, we show that a T ->C substitution in exon 9 (Y402H) of the complement factor H gene is strongly associated with susceptibility to age-related macular degeneration, the most common cause of blindness in the elderly. Frequency of the C allele was 0.61 in cases, versus 0.34 in age-matched controls (P<1x10(-24)). Genotype frequencies also differ markedly between cases and controls (chi2=112.68 [2 degrees of freedom]; P<1x10(-24)). A multiplicative model fits the data well, and we estimate the population frequency of the high-risk C allele to be 0.39 (95% confidence interval 0.36-0.42) and the genotype relative risk to be 2.44 (95% confidence interval 2.08-2.83) for TC heterozygotes and 5.93 (95% confidence interval 4.33-8.02) for CC homozygotes. PMID- 15895327 TI - The effect of a scheduled telephone intervention on outcome after moderate to severe traumatic brain injury: a randomized trial. AB - OBJECTIVE: To measure the effectiveness of a scheduled telephone intervention offering counseling and education to people with traumatic brain injury (TBI) on behavioral outcomes compared with standard follow-up at 1 year postinjury. DESIGN: Two-group randomized, prospective clinical trial throughout the first year after injury. SETTING: Subjects' homes via telephone in an urban-rural catchment area from a level I trauma center. PARTICIPANTS: Subjects (N=171; age range, 18-70 y) with a primary diagnosis of TBI who were discharged from an acute rehabilitation unit. They were randomly assigned to the telephone intervention (n=85) or to standard follow-up (n=86) groups at discharge. Of these, 79 participated in the intervention and completed the outcome assessment (3 withdrew; 3 were lost to follow-up), and 78 participated in usual care and completed the outcome assessment (8 were lost to follow-up). INTERVENTIONS: Subjects were randomly assigned to receive telephone calls at 2 and 4 weeks and 2, 3, 5, 7, and 9 months after discharge. The calls consisted of brief motivational interviewing, counseling, and education, plus facilitating usual care or usual care alone through follow-up appointments and therapy prescriptions. MAIN OUTCOME MEASURES: A composite outcome was used as the primary endpoint on an intent-to-treat basis. Secondary analyses were conducted with individual measures, including the FIM instrument, Disability Rating Scale, Community Integration Questionnaire, Neurobehavioral Functioning Inventory, Functional Status Examination, Glasgow Outcome Scale-Extended, Medical Outcomes Study 36-Item Short-Form Health Survey, Brief Symptom Inventory, EuroQol, and Modified Perceived Quality of Life scale. The primary analysis was a blocked Mann Whitney U test. RESULTS: At 1-year follow-up, those who had received scheduled telephone intervention fared significantly better on the primary composite outcome index ( P =.002). In addition, this group fared better on specific composites such as functional status ( P =.003) and quality of well-being ( P =.006). There were no significant differences on vocational status ( P =.08) or community integration status ( P =.13). CONCLUSIONS: Scheduled telephone counseling and education resulted in improved overall outcome, particularly for functional status and quality of well-being, when compared with usual outpatient care. Telephone counseling shows promise as a low-cost, widely available rehabilitation intervention for TBI. PMID- 15895328 TI - Increasing days at work using function-centered rehabilitation in nonacute nonspecific low back pain: a randomized controlled trial. AB - OBJECTIVE: To evaluate the effect of function-centered compared with pain centered inpatient rehabilitation in patients whose absence from work is due to chronic nonspecific low back pain (LBP). DESIGN: Single-blinded randomized controlled trial with follow-up assessments immediately after treatment and at 3 months. SETTING: Center for work rehabilitation in Switzerland. PARTICIPANTS: Patients with more than 6 weeks of work absence due to chronic nonspecific LBP (N=174; 137 men, 37 women; mean age +/- standard deviation, 42+/-8 y; mean sick leave before study, 6.5 mo). INTERVENTIONS: Function-centered treatment (FCT) (4h/d, 6d/wk, for 3 wk) consisted of work simulation, strength, endurance, and cardiovascular training. Pain-centered treatment (PCT) (2.5h/d, 6d/wk, for 3 wk) used a mini back school, individually selected passive and active mobilization, stretching, and low-intensity strength training. MAIN OUTCOME MEASURES: The number of days at work in 3 months after treatment, self-efficacy, lifting capacity, pain, mobility, strength, and global perceived effect. Effect sizes (ESs) (Cohen d ) were defined as small (ES range, 0.2-0.5), moderate (ES range, 0.5-0.8), and large (ES, >0.8). RESULTS: Groups were comparable at baseline. Moderate ESs for the FCT group versus PCT group were found for days at work (25.9 d vs 15.8d, ES=.36, P =.029), self-efficacy (5.9 points vs -7.4 points, ES=.55, P =.003), and lifting capacity (2.3 kg vs 0.2 kg, ES=.54, P =.004). CONCLUSIONS: Function-centered rehabilitation increases the number of work days, self efficacy, and lifting capacity in patients with nonacute nonspecific LBP. PMID- 15895329 TI - Effects of home strength training and stretching versus stretching alone after lumbar disk surgery: a randomized study with a 1-year follow-up. AB - OBJECTIVE: To assess the adherence to and effects of a 12-month combined strength and stretching home exercise regimen versus stretching alone, on patient outcome after lumbar disk surgery. DESIGN: Randomized controlled trial. SETTING: Departments of physical medicine and rehabilitation and orthopedics at a Finnish hospital. PARTICIPANTS: Patients (N=126) were randomized into either a combined strength training and stretching group (STG, n=65) or a control group (CG, n=61). INTERVENTION: The STG was instructed to perform strength training and both the STG and CG were instructed in the same stretching and stabilization exercises for 12 months. MAIN OUTCOME MEASURES: Pain on the visual analog scale (VAS), the Oswestry and the Million disability indexes, isometric and dynamic trunk muscle strength, mobility in the lumbar spine, and straight-leg raising were measured. RESULTS: The trial was completed by 71% and 77% of the patients from the STG and the CG, respectively. The mean strength training frequency decreased from 1.5 to 0.6 times a week in the STG during the intervention. The mean stretching frequency decreased from 3.7 to 1.6 times a week in both groups. Median back and leg pain varied between 17 and 23 mm (VAS), and the Million and Oswestry indices varied between 14 and 23 points 2 months postoperatively. No statistically significant changes took place in these outcome measures during the 12-month follow-up in both groups. The changes in isometric trunk extension favored the STG ( P =.016) during the first 2 months. However, during the whole 12-month training period, both dynamic and isometric back extension and flexion strength, as well as mobility of the spine and repetitive squat-test results, improved significantly in both groups, and no differences were found in any of the physical function parameters between the STG and CG. CONCLUSIONS: At the 12-month follow-up, no statistically significant changes were found in the physical function, pain, or disability measures between the groups. In the STG, training adherence with regard to training frequency and intensity remained too low to lead to specific training-induced adaptations in the neuromuscular system. Progressive loading, supervision of training, and psychosocial support is needed in long-term rehabilitation programs to maintain patient motivation. PMID- 15895330 TI - Moderate-heeled shoes and knee joint torques relevant to the development and progression of knee osteoarthritis. AB - OBJECTIVE: To determine if women's dress shoes with heels of just 1.5 in (3.8 cm) in height increases knee joint torques, which are thought to be relevant to the development and/or progression of knee osteoarthritis (OA) in both the medial and patellofemoral compartments. DESIGN: Randomized controlled trial. SETTING: A 3 dimensional motion analysis gait laboratory. PARTICIPANTS: Twenty-nine healthy young women (age, 26.7+/-5.0 y) and 20 healthy elderly adult women (age, 75.3+/ 6.5 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Peak external varus knee torque in early and late stance and prolongation of flexor knee torque in early stance. Three-dimensional data on lower-extremity torques and motion were collected during walking while (1) wearing shoes with 1.5-in high heels and (2) wearing control shoes without any additional heel. Data were plotted and qualitatively compared; major peak values and timing were statistically compared between the 2 conditions using paired t tests. RESULTS: Peak knee varus torque during late stance was statistically significantly greater with the heeled shoes than with the controls, with increases of 14% in the young women and 9% in the elderly women. With the heeled shoes, the early stance phase knee flexor torque was significantly prolonged, by 19% in the young women and by 14% in elderly women. Also, the peak flexor torque was 7% higher with the heeled shoe in the elderly women. CONCLUSIONS: Even shoes with moderately high heels (1.5 in) significantly increase knee torques thought to be relevant in the development and/or progression of knee OA. Women, particularly those who already have knee OA, should be advised against wearing these types of shoes. PMID- 15895331 TI - Are we selecting the right patients for stroke rehabilitation in nursing homes? AB - OBJECTIVE: To examine the effect of stroke rehabilitation in the nursing home on community discharge rates and functional status among patients stratified by propensity to receive rehabilitation. DESIGN: Retrospective cohort. SETTING: Medicaid-certified nursing homes (N=945) in Ohio. PARTICIPANTS: Patients with stroke (N=2013) admitted to an Ohio nursing home. INTERVENTION: Rehabilitation therapy services. MAIN OUTCOME MEASURES: The propensity to receive rehabilitation, used to adjust for selection bias, was calculated for each patient by using a logistic regression model. Community discharge and change in functional status, measured by using a crosswalk to the FIM instrument, were determined 3 months after admission. RESULTS: By 3 months after admission, 36.9% of the patients were discharged to the community, 16.6% had died, and 46.5% remained in the nursing home. The overall effect of rehabilitation on community discharge (relative risk [RR]=1.58; 95% confidence interval [CI], 1.33-1.85) was not homogeneous across subgroups stratified by propensity to receive rehabilitation. Patients less likely to receive rehabilitation, as measured by a lower propensity score, had a significant benefit in terms of community discharge (RR=1.65; 95% CI, 1.35-1.97), but those more likely to receive services did not (RR=1.21; 95% CI, 0.87-1.56). Among long-term nursing home residents, rehabilitation services were not associated with improved functional status. CONCLUSIONS: With respect to community discharge, patients who were less likely to receive rehabilitation therapy appear to receive greater benefit from rehabilitation services than those who were more likely to receive rehabilitation. This finding raises concerns about current selection practices for rehabilitation services. Research is needed to identify the patients most likely to benefit, especially in the present fiscally constrained reimbursement environment. PMID- 15895332 TI - Primary care practitioners' understanding of physiatric practice: effects on intention to refer. AB - OBJECTIVES: To learn what family practice and internal medicine physicians understand about the scope of practice of physical medicine and rehabilitation (PM&R) and to study what effect that understanding and various demographic variables have on their intention to refer to physiatrists. DESIGN: Survey-based. SETTING: National survey. PARTICIPANTS: One thousand internal medicine and family practice physicians were contacted, with 460 respondents. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Intention to refer patients to physiatrists using 13 case scenarios (10 appropriate referrals, 4 inappropriate referrals) and self-reported number of referrals per year associated with understanding of 7 skills of physiatrists analyzed by multiple logistic regression analyses. RESULTS: Although most respondents were likely to refer to physiatrists, a wide variation existed in the types of patients referred. Physicians with a greater understanding of the scope of physiatric practice were more likely to refer ( P =.003). Female physicians were more likely to refer than male physicians ( P =.003). CONCLUSIONS: There appears to be an association between an understanding of physiatric practice and primary care practitioners' willingness to refer to PM&R. Primary care physicians should be educated about the benefits of referring patients to physiatrists. PMID- 15895333 TI - Impairment and partial recovery of medical decision-making capacity in traumatic brain injury: a 6-month longitudinal study. AB - OBJECTIVE: To investigate empirically change in medical decision-making capacity (MDC) in patients with traumatic brain injury (TBI). DESIGN: Longitudinal study comparing control and TBI groups at hospitalization and at 6 months postinjury. SETTING: Inpatient brain injury rehabilitation unit. PARTICIPANTS: Twenty healthy controls and 24 patients with moderate to severe TBI. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: MDC was measured by using the Capacity to Consent to Treatment Instrument (CCTI). The CCTI evaluates performance on a series of 4 accepted consent abilities, or standards: S1 (evidencing/communicating choice), S3 (appreciating consequences), S4 (reasoning about treatment), and S5 (understanding the treatment situation and choices), and 1 experimental standard [S2] (making the reasonable treatment choice when the alternative choice is unreasonable). In addition, TBI patients were assigned 1 of 3 capacity outcomes (capable, marginally capable, incapable) for each standard. RESULTS: At hospitalization, TBI patients performed equivalently with controls on standards S1 and [S2] but significantly below controls on S3 ( P <.001), S4 ( P <.02), and S5 ( P <.001). At 6-month follow-up, TBI patients showed significant within-group improvement on these 3 standards (S3, S4, S5) but continued to fall significantly below controls on S3 ( P <.006) and S5 ( P <.001). A group by time interaction emerged on S5 ( P <.02). The TBI group showed increasing proportions of capable outcomes on all standards over the 6 months. CONCLUSIONS: Patients with TBI showed initial impairment and subsequent partial recovery of MDC over a 6-month period. Complex consent abilities of appreciation, reasoning, and understanding were significantly impaired in hospitalized acute TBI patients. At follow-up, TBI patients showed substantial recovery of reasoning and partial recovery of appreciation and understanding consent abilities. The study suggests the importance in the rehabilitation setting of serial evaluations of MDC in patients with TBI. PMID- 15895334 TI - Multidimensional assessment of acute confusion after traumatic brain injury. AB - OBJECTIVES: To describe the phenomenology of posttraumatic confusional state (PTCS) and to provide preliminary validation of a new procedure, the Confusion Assessment Protocol (CAP), for assessing PTCS. DESIGN: Criterion standard investigation. SETTING: Inpatient traumatic brain injury (TBI) rehabilitation program. PARTICIPANTS: Two consecutive series of patients (n=62, n=93) with TBI admitted for inpatient rehabilitation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Clinical diagnosis of delirium based on Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (DSM-IV) criteria, classification of posttraumatic amnesia (PTA) based on the Galveston Orientation and Amnesia Test (GOAT), and Disability Rating Scale score at time of rehabilitation hospital discharge. Results Agreement between the diagnosis of PTCS with the CAP and DSM-IV classification of delirium was 87%, and agreement between PTCS and PTA using GOAT criteria was 90%. Patients classified as in PTCS sustained more severe injuries and required longer rehabilitation stays. Confusion status was associated with poorer functional status at rehabilitation discharge. CONCLUSIONS: The CAP is a brief, structured, repeatable measure of multiple neurobehavioral aspects of PTCS. Confusion status as determined by CAP assessment contributed to prediction of outcome at rehabilitation discharge after adjustment for other potential predictors. PMID- 15895335 TI - Mental illness, traumatic brain injury, and medicaid expenditures. AB - OBJECTIVE: To estimate the rates of mental illness among Medicaid beneficiaries with traumatic brain injury (TBI) and associated Medicaid-paid expenditures. DESIGN: Retrospective claims-based calendar year data. SETTING: Claims data. PARTICIPANTS: Medicaid recipients with diagnosed TBI and mental illness who received Medicaid services in 4 states in 1995. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Annual expenditures for total, inpatient, and noninpatient services, as derived from Medicaid personal summary files. Mental illness and TBI were identified by using International Classification of Diseases, 9th Revision , Clinical Modification codes recorded in Medicaid claims. RESULTS: Of a total of 493,663 Medicaid recipients, 3641 (0.7%) were diagnosed with TBI in the 4 states. Significant demographic and racial differences were found in the rates of TBI; 18% of patients with TBI were diagnosed with serious mental illness. People with TBI in the age group 40 to 49 years were more likely to have a mental disorder. There were significant differences in estimated total, inpatient, and noninpatient expenditures between those with and without mental illness. In general, those with serious mental illness had higher Medicaid-paid expenditures than those without any mental illness. CONCLUSIONS: Psychiatric comorbidity in TBI increases the overall expenditures in this population. This increased cost is an important consideration in programming for those with TBI. PMID- 15895336 TI - Borg's rating of perceived exertion scales: do the verbal anchors mean the same for different clinical groups? AB - OBJECTIVE: To examine the interpretation of the verbal anchors used in the Borg rating of perceived exertion (RPE) scales in different clinical groups and a healthy control group. DESIGN: Prospective experimental study. SETTING: Rehabilitation center. PARTICIPANTS: Nineteen subjects with brain injury, 16 with chronic low back pain (CLBP), and 20 healthy controls. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Subjects used a visual analog scale (VAS) to rate their interpretation of the verbal anchors from the Borg RPE 6-20 and the newer 10-point category ratio scale. RESULTS: All groups placed the verbal anchors in the order that they occur on the scales. There were significant within group differences ( P >.05) between VAS scores for 4 verbal anchors in the control group, 8 in the CLBP group, and 2 in the brain injury group. There was no significant difference in rating of each verbal anchor between the groups ( P >.05). CONCLUSIONS: All subjects rated the verbal anchors in the order they occur on the scales, but there was less agreement in rating of each verbal anchor among subjects in the brain injury group. Clinicians should consider the possibility of small discrepancies in the meaning of the verbal anchors to subjects, particularly those recovering from brain injury, when they evaluate exercise perceptions. PMID- 15895337 TI - Late auditory and event-related potentials can be useful to predict good functional outcome after coma. AB - OBJECTIVE: To investigate whether late auditory and event-related potentials, and in particular N100 and mismatch negativity, together with clinical parameters, can help to predict good functional outcome in comatose patients. DESIGN: Prospective cohort study. SETTING: Hospital. PARTICIPANTS: Consecutively sampled comatose patients (N=346) whose etiologies of coma were stroke (125 patients), brain injury (96 patients), anoxia (64 patients), complication of neurosurgery (54 patients), and encephalitis (7 patients). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Glasgow Outcome Scale score at 1 year postonset. Patients in a minimally conscious state and those who awoke and died during the follow-up period were classified separately. RESULTS: Univariate analysis showed that all variables studied, except brainstem auditory evoked potentials, correlated significantly with functional outcome. Mismatch negativity showed the highest positive predictive value for good outcome. A validated model was obtained with multivariate logistic analysis, including pupillary light reflex, N100, mismatch negativity, etiology, and age. CONCLUSIONS: Late auditory and event-related potentials, and particularly N100 and mismatch negativity, provide strong prognostic factors for good functional outcome. Furthermore, these components may enhance the accuracy of prognosis when associated with other clinical parameters available at the early stage of coma. PMID- 15895338 TI - Effective rehabilitation for children and adolescents with brain injury: evaluating and disseminating the evidence. AB - OBJECTIVE: To develop and evaluate literature reviews of the effectiveness of rehabilitation interventions for children with brain injury. DESIGN: We wrote 6 research summaries, which were evaluated by study participants. SETTING: Community. PARTICIPANTS: A convenience sample that included 18 parents of children with brain injury, 18 service providers, and 12 insurance industry representatives. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Questionnaire designed for the evaluation of printed material. RESULTS: Few articles were found that focused on evaluation of rehabilitation interventions for children with brain injury. Study participants were representative of a broad range of educational and professional backgrounds. Before reading the research summaries, service providers reported greater familiarity with the topics than did parents and insurance representatives. Despite this finding, there were no significant between-group differences in the format, content, and impact ratings provided by the 3 participant groups. CONCLUSIONS: Research summaries written in a clear, straightforward manner are appropriate for people with different educational and professional backgrounds. Further research into the effectiveness of rehabilitation interventions is needed to support informed decision-making that results in the best outcomes for children with brain injury. PMID- 15895339 TI - Assessing mobility in children using a computer adaptive testing version of the pediatric evaluation of disability inventory. AB - OBJECTIVE: To assess score agreement, validity, precision, and response burden of a prototype computerized adaptive testing (CAT) version of the Mobility Functional Skills Scale (Mob-CAT) of the Pediatric Evaluation of Disability Inventory (PEDI) as compared with the full 59-item version (Mob-59). DESIGN: Computer simulation analysis of cross-sectional and longitudinal retrospective data; and cross-sectional prospective study. SETTING: Pediatric rehabilitation hospital, including inpatient acute rehabilitation, day school program, outpatient clinics, community-based day care, preschool, and children's homes. PARTICIPANTS: Four hundred sixty-nine children with disabilities and 412 children with no disabilities (analytic sample); 41 children without disabilities and 39 with disabilities (cross-validation sample). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Summary scores from a prototype Mob-CAT application and versions using 15-, 10-, and 5-item stopping rules; scores from the Mob-59; and number of items and time (in seconds) to administer assessments. RESULTS: Mob-CAT scores from both computer simulations (intraclass correlation coefficient [ICC] range, .94-.99) and field administrations (ICC=.98) were in high agreement with scores from the Mob-59. Using computer simulations of retrospective data, discriminant validity, and sensitivity to change of the Mob-CAT closely approximated that of the Mob-59, especially when using the 15- and 10-item stopping rule versions of the Mob-CAT. The Mob-CAT used no more than 15% of the items for any single administration, and required 20% of the time needed to administer the Mob-59. CONCLUSIONS: Comparable score estimates for the PEDI mobility scale can be obtained from CAT administrations, with losses in validity and precision for shorter forms, but with a considerable reduction in administration time. PMID- 15895340 TI - Health-related quality of life in childhood cerebral palsy. AB - OBJECTIVE: To describe the health-related quality of life (HRQOL) of a cross section of children with cerebral palsy (CP) using the Child Health Questionnaire. DESIGN: Survey of the parents or guardians of children with CP. SETTING: Outpatient clinics at a tertiary care children's hospital. PARTICIPANTS: Children (N=177; 98 boys, 79 girls; mean age +/- standard deviation [SD], 8.6+/ 4.2y) with CP were enrolled as a convenience sample. Study subjects were stratified by severity of CP using the Gross Motor Function Classification System. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: HRQOL scores, as determined by the Child Health Questionnaire-Parent Form 50. RESULTS: Parents reported reduced HRQOL for their children with CP. In the physical function domain, most children scored over 3 SDs below a normative sample, and scores in parental impact domains were at least 1 SD below the normative sample. Psychosocial domains were less impaired. Most of the effects on physical and parent impact domains were greater in children with more severe CP, although they were significant in most strata of severity. CONCLUSIONS: Children with CP have reduced HRQOL and the degree to which it is reduced is related to the severity of their CP. PMID- 15895341 TI - Visual disability variables. I: the importance and difficulty of activity goals for a sample of low-vision patients. AB - OBJECTIVE: To test the validity and reliability of latent trait measures estimated from ratings by low-vision patients of the importance and difficulty of selected activity goals. DESIGN: Validation of a telephone-administered functional assessment instrument using Rasch analysis of self-assessment ratings. SETTING: Telephone interviews of respondents in their homes. Participants Consecutive series of 600 outpatients with low vision. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Ratings of the importance and difficulty of achieving 41 activity goals. Person and item traits were measured with the Andrich rating scale model. Measurement validity and reliability were tested statistically by comparing response patterns and distributions with measurement model expectations. RESULTS: Patients could distinguish only 3 categories of importance and 4 categories of difficulty. The distributions of person and item measure fit statistics were consistent with 2 unidimensional constructs: value of independence estimated from importance ratings and visual ability estimated from difficulty ratings. However, 8 of 41 activity goals were poor estimators of value of independence and 7 of 41 activity goals were poor estimators of visual ability. Person measure distributions could be divided into 3 statistically distinct strata for estimates from both importance ratings and difficulty ratings. Item measure distributions could be divided into 21 strata for estimates from importance ratings and 7 strata for estimates from difficulty ratings. CONCLUSIONS: The 2 variables that define visual disability-value of independence and visual ability-are valid constructs that can be estimated accurately and reliably from patient ratings of the importance and difficulty of activity goals. PMID- 15895342 TI - Visual disability variables. II: The difficulty of tasks for a sample of low vision patients. AB - OBJECTIVES: To test the validity and reliability of measures of visual ability and to evaluate the relation between measurements made at the task level and measurements made at the goal level of a hierarchical model for visual disability. DESIGN: Validation of a telephone-administered functional assessment instrument using Rasch analysis on self-assessment ratings. SETTING: Telephone interviews of respondents in their homes. PARTICIPANTS: Consecutive series of 600 outpatients with low vision. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Ordinal ratings of the difficulty in performing a subset of 337 tasks. Measures of the visual ability of each patient and the required visual ability to perform each task were made using the Andrich rating scale model. Measurement validity and reliability were tested statistically by comparing response patterns and distributions to measurement model expectations. RESULTS: Results were consistent with a single visual ability construct. Patients' visual ability estimated from task difficulty ratings agreed with estimates from goal difficulty ratings ( r =.74); the difficulty of individual goals was equal to the weighted average of the difficulties of subsidiary tasks ( r =.79). However, conclusions from the Rasch analysis were not confirmed by principal components analysis of item residuals, which indicated that visual ability had a 2-dimensional structure, with 1 factor related to mobility and the other related to reading. Factor analysis on person measures estimated from subsets of functionally grouped items confirmed the 2-dimensional structure of visual ability. CONCLUSIONS: Our study results confirm the hierarchical structure of the Activity Breakdown Structure model and show how the individualized Activity Inventory can produce measures of limitations in functional vision. PMID- 15895343 TI - Coronary artery disease and risk factors in people with posttraumatic vision loss. AB - OBJECTIVE: To examine the prevalence of coronary artery disease (CAD) and its risk factors in people with posttraumatic vision loss (PTVL). DESIGN: Cross sectional, controlled study. SETTING: The general community. PARTICIPANTS: Study groups included 82 subjects with PTVL, 49 siblings, 58 blind subjects with retinitis pigmentosa (RP), and the general population in Israel. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Sociodemographic and biomedical data collected by using a structured questionnaire and medical records. RESULTS: The prevalence of CAD among subjects with PTVL (24%) was 2 to 3 times higher than the control groups ( P <.001). However, the prevalence of the CAD risk factors in these subjects was similar to or lower than those in the control groups. For example, significantly fewer subjects with PTVL were physically inactive (16%) than patients with RP (55%, P <.01). The only variable that was significantly associated with CAD prevalence was the cause of blindness-that is, trauma versus disease; the odds of having CAD after traumatic vision loss was 3.75 times higher than after RP. CONCLUSIONS: People with PTVL exhibit elevated rates of risk for CAD similar to those of other groups with physical disability. The traumatic injury that caused vision loss might be an important factor underlying that risk. PMID- 15895344 TI - In vitro maturation potential of monocyte-derived dendritic cells is impaired in patients with spinal cord injury: a case-control study. AB - OBJECTIVE: To determine whether the T-cell reactivity and maturation potential of dendritic cells are impaired in subjects with spinal cord injury (SCI). DESIGN: Cross-sectional, case-control study. SETTING: University hospital in Taiwan. PARTICIPANTS: Thirty male SCI subjects, including 14 with paraplegia and 16 with tetraplegia, and 30 age- and sex-matched healthy controls. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: CD4+ and CD8+ T-cell reactivity was assessed by lymphoproliferative response (LPR) and CD69 expression in response to stimulation with antigens, anti-CD3 monoclonal antibody, and mitogen. Dendritic cell maturation potential was assessed by phenotypic (CD80 and CD83 expression in stimulated monocyte-derived dendritic cells) and functional (the LPR of alloreactive T cells in mixed leukocyte reactions) analysis. RESULTS: The potential of phenotypic and functional maturation of dendritic cells of subjects with SCI was significantly impaired compared with healthy controls, and that potential was worse for tetraplegic patients than for paraplegic patients. However, no significant difference was found in T-cell responses of CD4+ and CD8+ subsets between subjects with SCI and healthy controls. CONCLUSIONS: Impaired maturation potential of dendritic cells is a novel defect in innate immunity in people with SCI. PMID- 15895345 TI - Sex education, relationships, and sexuality in young adults with spina bifida. AB - OBJECTIVES: To assess the adequacy of sex education and to determine the incidence of various difficulties encountered in relationships and sexual contact by young adults who have spina bifida (SB) with and without hydrocephalus (HC) in the Netherlands. DESIGN: Cross-sectional. SETTING: Community. PARTICIPANTS: Patients with SB occulta or aperta (N=157; 41% male; mean age, 20.8 y; age range, 16-25 y). Interventions Not applicable. MAIN OUTCOME MEASURES: Structured interview on sex education, relationships, sexual activities, and sexual functioning. RESULTS: Although sex education had been provided to almost all patients, fewer than a quarter received information specific to people with SB. Of all patients, 25% had a partner, 70% desired sexual contact, 47% had had sexual contact, and 22% had had sexual intercourse during the last year. Only 52% were satisfied with their present sex life. Incontinence and lack of self confidence were important obstacles. Compared with patients without HC (HC-), patients with HC (HC+) less often had a partner, were sexually less active, and more often had problems with sexual functioning. Predictors of not having sexual contact were having HC and being male. CONCLUSIONS: Relationships and sexuality are important for young adults with SB. HC+ patients are less active and perceive more problems than HC- patients. Counseling in relationships and sexuality should be part of the regular care for this group. PMID- 15895346 TI - Prognostic value of pinprick preservation in motor complete, sensory incomplete spinal cord injury. AB - OBJECTIVE: To assess sacral and lower-extremity pinprick preservation as prognostic indicators for ambulation in motor complete, sensory incomplete spinal cord injury (SCI). DESIGN: Retrospective analysis. SETTING: Twenty-eight tertiary care centers in the United States and Canada. PARTICIPANTS: Subjects (N=131; mean age, 31.6y) with motor complete, sensory incomplete SCI. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Ambulation at 26 and 52 weeks postinjury (modified Benzel scale). RESULTS: A higher percentage of subjects with sacral pinprick preservation at baseline were ambulating at 26 (39.4% vs 28.3%) and 52 weeks (53.6% vs 41.5%). This finding did not reach statistical significance. The presence of sacral pinprick preservation at 4 weeks postinjury was significant for predicting ambulation at 52 weeks postinjury (36.0% vs 4.4%, P =.011) and approached significance at 26 weeks (15.2% vs 0.0%, P =.056). Significant differences in ambulation rates were also observed between subjects, based on the presence of baseline lower-extremity pinprick preservation (>/=50% of lower extremity L2-S1 dermatomes) at both 26 (50.0% vs 28.8%, P =.048) and 52 weeks (66.7% vs 40.3%, P =.023) after injury. CONCLUSIONS: Baseline lower-extremity pinprick preservation and sacral pinprick preservation at 4 weeks postinjury are associated with an improved prognosis for ambulation. PMID- 15895347 TI - Validation of a new lower-extremity motor coordination test. AB - OBJECTIVE: To determine the test-retest reliability and construct validity of a new lower-extremity motor coordination test, the Lower Extremity MOtor COordination Test (LEMOCOT). DESIGN: To test reliability, subjects with impairments in at least 1 lower extremity were evaluated twice by the same evaluator. To test construct validity, the LEMOCOT scores obtained from subjects who had had a stroke were correlated with physical, functional, cognitive, and perceptual tests. SETTING: Geriatric day hospital and functional intensive rehabilitation unit. PARTICIPANTS: In the reliability test, 29 people (mean age, 69.6y; range, 28-87y); in the construct validity, 144 people who recently had had a stroke. INTERVENTION: Not applicable. MAIN OUTCOME MEASURES: In addition to the LEMOCOT, the following measures were used for construct validity: the Fugl-Meyer Assessment (motor function), Berg Balance Scale, 5-m walking test, 2-minute walking test, Functional Autonomy Measurement System, Modified Mini-Mental State Examination, and Motor-Free Visual Perceptual Test. RESULTS: Intraclass correlation coefficients (ICCs) indicated that test-retest reliability is good (right-side ICC=.88; left-side ICC=.83). The construct validity of the LEMOCOT was demonstrated by obtaining high correlations with physical and functional tests ( r range, .62-.79; P <.001) and no correlations with cognitive ( r =.11, P =.20) or visual perceptual tests ( r =.15, P =.08) and by discriminating between subjects discharged to long-term care versus other living environments ( P <.001). CONCLUSIONS: The LEMOCOT is a simple lower-extremity motor coordination test that showed good test-retest reliability and construct validity. It can be used in clinical and research settings, specifically with people who have had a stroke. Other studies should be carried out to confirm its psychometric properties. PMID- 15895348 TI - The effect of external rhythmic cues (auditory and visual) on walking during a functional task in homes of people with Parkinson's disease. AB - OBJECTIVES: To evaluate (1) the influence of rhythmic cues on gait interference during a functional activity and (2) the relationship of clinical symptoms to gait interference. DESIGN: Repeated-measures study. SETTING: Participants' homes. PARTICIPANTS: Twenty subjects with idiopathic Parkinson's disease (PD) and a control group of 10 age-, sex-, and education-matched subjects. INTERVENTIONS: Subjects performed a simple functional task that included a walking component and a dual-motor task. The functional task was performed with and without external rhythmic (auditory and visual) cues. MAIN OUTCOME MEASURES: Walking speed, mean step length, and step frequency were compared during trials of the tasks. In addition, tests of cognitive executive function (Hayling and Brixton tests), anxiety and depression (Hospital Anxiety and Depression Scale), and fatigue (Multidimensional Fatigue Inventory) were undertaken. RESULTS: The use of auditory cues during a dual task involving gait reduced the interference effect on the task; significant increases in step length were observed in PD subjects ( P =.018), representing an increase of 19%. CONCLUSIONS: External auditory cues may be useful in reducing interference and maintaining gait performance during more complicated functional activities. Clinical symptoms, such as depression and fatigue, could influence the ability to focus attention and may increase gait interference during the performance of complex tasks, with subsequent implications for functional walking and safety. PMID- 15895349 TI - Quantitative gait analysis in Parkinson's disease: comparison with a healthy control group. AB - OBJECTIVE: To compare gait parameters in Parkinson's disease (PD) during the on phase of medication cycle with those of healthy elderly control subjects. DESIGN: A group-comparison study. SETTING: Gait analysis laboratory of a university hospital. PARTICIPANTS: Fifteen patients with PD and 9 healthy elderly controls. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Spatiotemporal, kinematic, and kinetic gait parameters. RESULTS: The PD spatiotemporal results showed a significant reduction in step length and walking velocity compared with controls. In the kinematics, the major feature of the PD group was a markedly reduced ankle plantarflexion excursion (at 50%-60% of the gait cycle). Most important, the kinetics showed reduced ankle push-off power and hip pull-off power. Unlike the control subjects, the patients with PD did not show any correlation between ankle generation (push-off) power and stride length ( r =.19) or with gait speed ( r =.29). Correction for walking velocity did not result in significant changes in the kinetics between the groups. CONCLUSIONS: Reduced ankle (push-off) power generation and reduced hip flexion (pull-off) power persisted in PD gait despite being tested in the on-phase of the medication cycle. Lack of a correlation between ankle and hip power generation and walking velocity suggests that peripheral and central factors contribute to lack of forward progression. Patients with PD may benefit from intervention strategies that correct the kinematic and the kinetic gait components. PMID- 15895350 TI - Validity and reliability of an internet-based temporal gait assessment tool with healthy adults: a pilot study. AB - OBJECTIVES: To assess in healthy adults the validity and the inter- and intrarater reliability of the Internet-based Shaw Gait Assessment (SGA). DESIGN: Concurrent test-retest reliability and validity study with participants, 4 raters, and the Elite motion analysis system (used as the criterion standard). SETTING: Motion analysis laboratory in a university physical therapy department. PARTICIPANTS: Convenience sample of 16 healthy men and women (age range, 28-53y). INTERVENTION: Each subject performed 2 consecutive walks "at a comfortable pace" on a 6-m walkway. A video camera from the Elite motion analysis system filmed reflective markers, which were attached to subjects' shoes, and the reflective markers provided the criterion standard. Four raters simultaneously recorded each walk by using laptop computers and the SGA. MAIN OUTCOME MEASURES: Paired t test (5% level) for average differences between each test and retest for raters and the Elite; Pearson correlations, limits of agreement, and coefficients of variation (CVs) for validity of the tool; intraclass correlation coefficients (ICCs) for inter- and intrarater reliability. RESULTS: Pearson product moment correlation coefficients between each of the raters and the Elite ranged from .92 to .95 for speed, from .85 to .97 for cadence, from .87 to .92 for step length, from .61 to .84 for left advance limb time, and from .68 to .83 for right advance limb time. Pooled CVs for all variables were below 8% for all raters and the Elite. Pooled ICCs for intrarater reliability were .89 for speed, .99 for cadence, .84 for step length, .76 for left limb advance time, and .84 for right limb advance time. Interrater ICCs were .89 for speed, .82 for cadence, .76 for step length, .66 for left limb advance time, and .81 for right limb advance time. CONCLUSIONS: The SGA is a valid and reliable tool for several key temporal measures of gait in a healthy adult population. PMID- 15895351 TI - Thermal pain perception after aerobic exercise. AB - OBJECTIVE: To examine thermal pain perception before, 5 minutes after, and 30 minutes after 30 minutes of treadmill exercise at 75% of maximal oxygen uptake (V o 2 max). DESIGN: Repeated-measures. SETTING: Sports science laboratory. PARTICIPANTS: Convenience sample of 14 healthy male and female volunteers (mean age +/- standard deviation, 32+/-3y). INTERVENTIONS: Sensory thresholds, pain thresholds, and pain ratings to hot and cold stimuli were measured before and after 30 minutes of treadmill exercise at 75% of V o 2 max. The hot and cold stimuli were delivered by using a thermode placed on the thenar eminence of the nondominant hand. Thermal sensory and pain thresholds were determined during continuous ramps in temperature of the thermode. MAIN OUTCOME MEASURES: Pain ratings were measured on a visual analog scale at 10-second intervals over 2 minutes of thermal pain stimulation. RESULTS: There were no significant changes in thermal sensitivity, pain thresholds, or pain ratings for either heat or cold after 30 minutes of exercise at 75% of V o 2 max. CONCLUSIONS: Pain perception to thermal stimuli was unaltered after 30 minutes of exercise at 75% of V o 2 max, an intensity and duration of exercise previously shown to alter pain perception to electric and mechanical stimuli. PMID- 15895352 TI - Comparison of cardiovascular responses between upright and recumbent cycle ergometers in healthy young volunteers performing low-intensity exercise: assessment of reliability of the oxygen uptake calculated by using the ACSM metabolic equation. AB - OBJECTIVES: To clarify (1) differences in cardiovascular response during low intensity exercise in the upright versus the recumbent position, and (2) whether the oxygen uptake (V o 2 ) calculated by the American College of Sports Medicine (ACSM) metabolic equation reflects the actual V o 2 at low-intensity testing. DESIGN: Repeated-measures comparison study. SETTING: University research laboratory. PARTICIPANTS: Thirty-one healthy, young volunteers (age, 23+/-2y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Blood pressure, rate pressure product (RPP), V o 2 , oxygen pulse, carbon dioxide output (V co 2 ), and ventilatory equivalent (V e ) were measured during graded exercise testing using upright and recumbent cycle ergometers. The estimated V o 2 was calculated by using the ACSM metabolic equation. RESULTS: Systolic blood pressure, RPP, V o 2 , oxygen pulse, V co 2 , and V e at 15 or 30W were significantly higher in the recumbent position than in the upright one ( P <.05), however, no significant differences were observed at 50 and 70W. The estimated V o 2 during exercise was significantly higher than the actual one, at every level of intensity, from 15 to 70W ( P <.05). CONCLUSIONS: Cardiovascular responses should be carefully monitored even during low-intensity exercise using a recumbent cycle ergometer. The V o 2 estimated using the ACSM metabolic equation did not reflect the actual V o 2 during low-intensity exercise at 70W or less. PMID- 15895353 TI - Metabolic syndrome in neuromuscular disease. AB - OBJECTIVES: To test the hypotheses that (1) people with neuromuscular disease (NMD) have multiple risk factors for cardiovascular disease and diabetes and (2) these risk factors worsen over time. DESIGN: Longitudinal testing with average 2.5-year follow-up. SETTING: Human performance laboratory of a university. PARTICIPANTS: Eleven ambulatory volunteers with slowly progressive NMD and 8 able bodied controls, group-matched for age and body mass index (BMI) at baseline. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Percentage of body fat (%BF), physical activity, energy expenditure, blood lipids and glucose, and blood pressure. RESULTS: At baseline, NMD subjects were more obese (37%BF vs 34%BF, respectively) and more sedentary than the controls, spending less time in total activity (144 min/d vs 214 min/d) and in exercise (11 min/d vs 45 min/d). The NMD group also had numerous cardiovascular and metabolic risk factors, with low high density lipoprotein cholesterol, high BMI, and high triglyceride being the most common. Additionally, 55% of the NMD group satisfied the criteria for metabolic syndrome, versus 0% in the control group. Most parameters did not significantly worsen during the average 2.5-year follow-up period in either group. CONCLUSIONS: People with NMD are at high risk for developing chronic diseases resulting from obesity and a sedentary lifestyle. Intervention studies aimed at reducing their risk for such chronic diseases are warranted. PMID- 15895354 TI - Strengthening of partially denervated knee extensors using percutaneous electric stimulation in a young man with spinal cord injury. AB - OBJECTIVE: To evaluate the effects of percutaneous electric stimulation on knee extensor strength and muscle hypertrophy, gait, and energy cost of walking in a young man with partial denervation of the knee extensors. DESIGN: One-way repeated measures. SETTING: Pediatric orthopedic hospital. PARTICIPANT: A man in his early twenties, who had an L2 American Spinal Injury Association class D spinal cord injury, presented with strength deficits in his left knee extensors and reported falling frequently. When walking, his left knee remained locked in extension throughout stance. Electromyographic testing revealed chronic denervation and reinnervation changes. INTERVENTION: Because of sensory difficulties with surface stimulation, a percutaneous electrode was surgically implanted near the femoral nerve. The subject exercised isometrically with a research grade stimulator for 1 hour a day until his strength plateaued. MAIN OUTCOME MEASURES: Quadriceps femoris strength and hypertrophy, gait, and energy cost of walking were recorded preintervention, every 2 months during the strengthening phase, and 2 months after withdrawal. RESULTS: Voluntary isometric torque improved from 7 to 14.8Nm (112%) and decreased to 8.5Nm after stimulation was withdrawn. Mean circumferential measures of the thigh improved from 12.3 to 13.5cm (9.8%) and then decreased to 13.1cm. Gait kinematics and kinetics were unchanged, although the subject reported greater stability in his left knee and fewer falls. CONCLUSIONS: The study indicates that percutaneous electric stimulation could be used to strengthen partially denervated muscle and to affect function. However, gains in strength may not be maintained once treatment is withdrawn. PMID- 15895355 TI - Factors affecting the burden on caregivers of stroke survivors in South Korea. AB - OBJECTIVE: To elucidate the factors related to the caregiver burden in stroke survivors in Seoul, Korea. DESIGN: Cross-sectional study. SETTING: Outpatient clinic of a university hospital in Korea. PARTICIPANTS: We randomly selected 340 stroke patients and evaluated them for 3 years poststroke. Among them, 147 caregivers and patients were finally interviewed. INTERVENTIONS: Patients' demographic data, neurologic symptoms, depression, emotional incontinence, and Modified Rankin Scale (MRS) score were assessed. In addition, primary caregivers' demographic data, hours of care per day, relationship with the patients, health status, anxiety and depression, and the presence of alternative caregivers were assessed. MAIN OUTCOME MEASURES: Caregiver burden was evaluated with the Sense of Competence Questionnaire, which has 3 subdomains with a total of 27 items. RESULTS: Caregiver burden score was high as compared with reports from Western society. Patient factors related to the burden scores were being unemployed ( P <.05), the presence of diabetes mellitus ( P <.05), aphasia ( P <.01), dysarthria ( P <.05), dysphagia ( P <.01), cognitive dysfunction ( P <.01), severe MRS score ( P <.01), emotional incontinence ( P <.05), and depression ( P <.01). Caregiver factors related to high burden scores were being female ( P <.05), unemployment ( P <.01), being daughters-in-law ( P <.01), anxiety ( P <.01), depression ( P <.01), and long caregiving hours ( P <.05). On multivariate analysis, the most significant predictor for overall caregiver burden was caregiver anxiety followed by patient MRS score and caregivers being daughters-in-law. Caregiver depression, current employment status, and patient depression were also factors related to their burden. CONCLUSIONS: Caregiver anxiety, patient physical deficits, and caregivers being daughter-in-law are important factors related to the caregiver burden in our population. Strategies to reduce caregiver burden should be developed based on understanding of these factors. PMID- 15895356 TI - Use of percutaneous endoscopic gastrostomy feeding tubes and functional recovery in stroke rehabilitation: a case-matched controlled study. AB - OBJECTIVE: To compare the morbidity, mortality, and functional recovery of patients who require percutaneous endoscopic gastrostomy (PEG) placement for the management of dysphagia after stroke. DESIGN: Retrospective case-matched controlled study. SETTING: Acute stroke rehabilitation inpatient unit. PARTICIPANTS: Patients (N=193) who were admitted for stroke rehabilitation with a PEG tube in place from January 1, 1993, to December 31, 2002, were matched with 193 case controls without PEG. Patients and controls were within 90 days of stroke onset, and were matched for age, sex, type of stroke, FIM instrument score, duration from onset to stroke unit admission, and year of admission. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Length of rehabilitation hospital stay, improvement in FIM scores, FIM efficiency score, need for transfer back to acute care hospital, diagnosis for which transfer was required, final discharge destination, and survival status. RESULTS: Significant differences were found between the 2 groups, PEG versus control, respectively, in the following variables: FIM efficiency (.42+/-.57 vs .56+/-.55, P =.016); need for transfer back to acute hospital (58/193 vs 23/193, P =.001); and survival status dead/alive (14/179 vs 3/190, P =.006). Nonsignificant differences were as follows: length of rehabilitation hospital stay (46.9+/-24.8d vs 43.3+/-19.7d, P =.11), improvement in total FIM score from admission to discharge (16.9+/-17.9 vs 21.0+/-15.5, P =.72), and final discharge destination home/institutional care (96/83 vs 101/89, P =.93). Pneumonia was the most frequent reason for transfer to acute care for patients with PEG. CONCLUSIONS: Patients who meet criteria for admission to a stroke rehabilitation unit and who have a PEG in place are at increased risk for medical complications and death. Those who survive, however, show similar functional recovery and rate of home discharge as case-matched controls. PMID- 15895357 TI - Erdheim-Chester disease: the effect of bisphosphonate treatment--a case report. AB - Erdheim-Chester disease is a distinctive pathologic and radiographic entity characterized by bilateral symmetric sclerosis of the diametaphyseal regions of long bones and infiltration of foamy lipid-laden histiocytes. It is a rare histiocytic disease of unknown etiology that is characterized pathologically by xanthogranulomatous infiltrates of multiple organs. We present a patient in her early sixties with bilateral mild knee and leg pain. The patient showed a typical bilateral symmetric medullary sclerosis at the diametaphyseal portions of long bones of the lower extremity. The diagnosis was confirmed by a bone biopsy, and bisphosphonate (alendronate, 70 mg/wk) was given to the patient. After 9 months of treatment, biochemical markers of bone turnover, which were high at baseline, decreased to normal ranges. However, the radiographs showed that bone lesions had changed to lytic lesions. We propose use of bisphosphonates, such as alendronate, to decrease the biochemical markers of bone turnover. But we suggest that it is premature to conclude that bisphosphonates have any effect on lytic lesions and the progression of the disease as shown by changes in radiographs. Further studies with long-term follow-up and ultrastructural evaluation are needed. PMID- 15895358 TI - Magnetic resonance neurography diagnosed brachial plexitis: a case report. AB - Idiopathic brachial plexitis is a rare disorder presenting with pain and weakness in the shoulder girdle and upper extremity. Idiopathic brachial plexitis can mimic other conditions that cause acute pain and weakness around the shoulder, and its diagnosis can be challenging. There is no special test for the diagnosis of idiopathic brachial plexitis, although electromyography may be useful. In this case of idiopathic brachial plexitis, we present magnetic resonance neurography findings for the first time. PMID- 15895359 TI - A novel approach to prevent repeated catheter migration in a patient with a baclofen pump: a case report. AB - We report a novel solution to a problem of repeated catheter migration that may aid others caring for patients with catheter migration problems. Catheter migration is a frequently reported complication of intrathecal drug delivery systems. We report on an ambulatory patient with a baclofen pump for control of spasticity due to cerebral palsy and dystonia; the patient suffered repeated episodes of catheter migration. The ultimate solution to the migrations was to place a pediatric pump in the lower thoracic, paraspinal region with the catheter entering the thoracic spine directly adjacent to the pump, thereby minimizing the differential motion between the pump and the spine, which was thought to be the cause of the repeated migration. This solution has not been previously described. Paraspinal pump placement may eliminate repeated catheter migration for patients with intrathecal drug pumps. PMID- 15895360 TI - Assessment of verticality perception by a rod-and-frame test: preliminary observations on the use of a computer monitor and video eye glasses. AB - A computer-based version of the rod-and-frame test for verticality perception has been developed. The task tested verticality perception of subjects by using the computer mouse to rotate a bright rod to a vertical orientation within a square frame that was tilted through a range of angles from 30 degrees counterclockwise to 30 degrees clockwise. In 16 healthy subjects, errors from the true vertical were generated in the direction of the frame tilt. The errors reached statistical significance ( P <.05) when the frame was rotated by 10 degrees in either direction. In a second study, the images were presented to the subjects who wore a pair of video eyeglasses. The pattern of errors generated by rotation of the frame were similar but were of larger amplitude than those produced in the first experiment and reached statistical significance with frame angles of 10 degrees and 20 degrees in both directions. These preliminary investigations suggest that the computer-based rod-and-frame test may provide a more convenient alternative to the mechanical rod-and-frame test for verticality perception. PMID- 15895361 TI - Design and validation of a methodology using the International Classification of Diseases, 9th Revision, to identify secondary conditions in people with disabilities. AB - OBJECTIVE: To design and validate a methodology that identifies secondary conditions using International Classification of Disease, 9th Revision (ICD-9) codes. DESIGN: Secondary conditions were identified through a literature search and a survey of Washington State physiatrists. These conditions were translated into ICD-9 codes and this list was then validated against a national sample of Medicare survey respondents with differing levels of mobility and activities of daily living (ADL) disability. SETTING: National survey. PARTICIPANTS: Participants (N=9731) in the 1999 Medicare Current Beneficiary Survey with no, mild, moderate, and severe mobility and ADL disability. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Percentage of survey respondents with a secondary condition. The secondary conditions were grouped into 4 categories: medical, psychosocial, musculoskeletal, and dysphagia related (problems associated with difficulty in swallowing). RESULTS: Our literature search and survey of 26 physiatrists identified 64 secondary conditions, including depression, decubitus ulcers, and deconditioning. Overall, 70.4% of all survey respondents were treated for a secondary condition. We found a significant relation between increasing mobility as well as ADL disability and increasing numbers of secondary conditions (chi 2 test for trend, P <.001). This relation existed for all categories of secondary conditions: medical (chi 2 test for trend, P <.001), psychosocial (chi 2 test for trend, P <.001), musculoskeletal (chi 2 test for trend, P <.001), and dysphagia related (chi 2 test for trend, P <.001). CONCLUSIONS: We created a valid ICD-9-based methodology that identified secondary conditions in Medicare survey respondents and discriminated between people with different degrees of disability. This methodology will be useful for health services researchers who study the frequency and impact of secondary conditions. PMID- 15895362 TI - Negative regulation of melanogenesis by phospholipase D1 through mTOR/p70 S6 kinase 1 signaling in mouse B16 melanoma cells. AB - Melanogenesis is a principal parameter of differentiation in melanocytes and melanoma cells. Our recent study has demonstrated that phospholipase D1 (PLD1) regulates the melanogenic signaling through modulating the expression of tyrosinase, the rate-limiting step enzyme in the melanin biosynthesis. The current study was designed to gain more insight into the involvement of PLD1 in the regulation of melanogenesis. To investigate the role of PLD1, we examined the effect of knockdown of endogenous PLD1 by small interference RNA (siRNA) on melanogenesis in B16 melanoma cells. It was shown that the melanin synthesis was induced in PLD1-knockdowned cells, and also that the level of melanin synthesis was well correlated with increases in expression level of tyrosinase and its related proteins (Tyrp1 and Dct). Furthermore, the reduction of expression levels of PLD1 by siRNA transfection was accompanied by diminution of ribosomal S6 kinase 1 (S6K1) phosphorylation. The activity of mammalian target of rapamycin (mTOR) is essential for phosphorylation of S6K1 and the treatment malanoma cells with rapamycin, a potent inhibitor of mTOR effectively induced melanogenesis. The results obtained here provide possible evidence that PLD1 exerts a negative regulatory role in the melanogenic process through mTOR/S6K1 signaling. PMID- 15895363 TI - Expression profile of the RNA-binding protein gene hermes during chicken embryonic development. AB - The hermes gene encodes an RNA-binding protein containing an RNA-recognition motif. Its expression has been described previously in Xenopus and in the developing heart of very young chicken embryos. We have analyzed the expression of cHermes in later heart development, where expression is maintained in the myocardium, and also in previously undescribed sites. cHermes expression first appears in the somites in the first terminally differentiated myocytes of both the epaxial and the hypaxial myotome. Expression is also seen in the primordium of the allantois and continues in the developing allantoic sac. cHermes expression in the pronephric and mesonephric kidneys coincides temporally and spatially with the appearance of the vascular components of the glomeruli. In addition, cHermes expression was seen in the mesoderm of the gut and in the notochord. PMID- 15895364 TI - Properties of Mg(2+)-dependent cation channels in human leukemia K562 cells. AB - The endogenous Mg(2+)-inhibited cation (MIC) current was recently described in different cells of hematopoietic lineage and was implicated in the regulation of Mg2+ homeostasis. Here we present a single channel study of endogenously expressed Mg(2+)-dependent cation channels in the human myeloid leukemia K562 cells. Inwardly directed unitary currents were activated in cell-attached experiments in the absence of Ca2+ and Mg2+ in the pipette solution. The current voltage (I-V) relationships displayed strong inward rectification and yielded a single channel slope conductance of approximately 30 pS at negative potentials. The I-V relationships were not altered by patch excision into divalent-free solution. Channel open probability (P(o)) and mean closed time constant (tau(C)) were strongly voltage-dependent, indicating that gating mechanisms may underlie current inward rectification. Millimolar concentrations of Ca2+ or Mg2+ applied to the cytoplasmic side of the membrane produced slow irreversible inhibition of channel activity. The Mg(2+)-dependent cation channels described in this study differ from the MIC channels described in human T-cells, Jurkat, and rat basophilic leukemia (RBL) cells in their I-V relationships, kinetic parameters and dependence on intracellular divalent cations. Our results suggested that endogenously expressed Mg(2+)-dependent cation channels in K562 cells and the MIC channels in other hematopoietic cells might be formed by different channel proteins. PMID- 15895365 TI - RBMX gene is essential for brain development in zebrafish. AB - The human RBMX gene was discovered recently through its homology to the spermatogenesis candidate gene RBMY. Its position on the human X chromosome suggests that it may be involved in X-linked mental retardation syndromes. However, to date there is scant information on the in vivo role of RBMX. To address this issue, we have isolated a zebrafish rbmx orthologue and characterized its embryonic expression pattern. Zebrafish rbmx is maternally expressed and then widely expressed in the embryo up to 24 hr postfertilization. In later stages of embryonic development, rbmx transcripts are localized predominantly in the brain, branchial arches, and liver primordium. The function of rbmx during embryonic development was examined by the use of an antisense morpholino targeting rbmx. The rbmx-morphants displayed an underdeveloped head and eyes, reduced body size, defective somite patterning, and absence of jaws. Furthermore, in the absence of functional rbmx, expression of specific markers for the fore- and hindbrain (otx2, krox20) was severely reduced. These studies demonstrate for the first time that rbmx is required for normal embryonic development, in particular of the brain, consistent with a role in X-linked mental retardation. PMID- 15895366 TI - Evidence for a role for anti-Mullerian hormone in the suppression of follicle activation in mouse ovaries and bovine ovarian cortex grafted beneath the chick chorioallantoic membrane. AB - The first critical transition in follicular development, the activation of primordial follicles to leave the pool of resting follicles and begin growth, is poorly understood, but it appears that the balance between inhibitory and stimulatory factors is important in regulating the exodus of follicles from the resting pool. There is evidence that anti-Mullerian hormone (AMH; also known as MIS) inhibits follicle activation in mice, but whether it plays a similar role in non rodent species is not known. When pieces of bovine ovarian cortex, rich in primordial follicles, are cultured in serum-free medium, most follicles initiate growth, but when cortical pieces are grafted beneath the chorioallantoic membrane (CAM) of chick embryos, follicle activation does not occur. Since embryonic chick gonads of both sexes produce and secrete high levels of AMH, the hypothesis that the AMH in the chick circulation inhibits follicle activation was tested. In Experiment 1, whole newborn mouse ovaries were grafted beneath the CAM (placed "in ovo") or cultured in vitro for 8 days. In vitro (or after 8 days in vivo) follicles activated and proceeded to the primary or secondary stage, but activation was suppressed in ovo. This inhibition was reversed if ovaries were removed from beneath the CAM and cultured in vitro. In contrast, when ovaries from mice null mutant for the AMH type II receptor were CAM-grafted in Experiment 2, follicle activation occurred in a similar fashion to activation in vitro. This finding strongly implicates AMH as the inhibitor of follicle activation in ovo. Since chick embryonic gonads are the source of circulating AMH, chicks were gonadectomized in Experiment 3, prior to grafting of pieces of bovine ovarian cortex beneath their CAMs. Bovine primordial follicles activated in the gonadectomized chicks, similar to the results for mice lacking the AMH type II receptor. Taken together these experiments provide strong evidence that AMH is the inhibitor of mouse follicle activation present in the circulation of embryonic chicks and provide indirect, and hence more tentative, evidence for AMH as an inhibitor of bovine follicle activation. PMID- 15895367 TI - Opposing roles of two isoforms of the Prx1 homeobox gene in chondrogenesis. AB - The Prx1 homeobox gene is critical for cartilage and bone development as suggested by previous expression studies and demonstrated by gene targeting. However, neither approach assessed the individual roles of the two isoforms Prx1a and Prx1b. In this study, Western blot analysis demonstrates that, in the early stages of chondrogenesis, during mesenchymal condensation, only Prx1a is expressed. Higher level Prx1b expression is concomitant with the formation of a defined perichondrium. Prx1a overexpression in limb micro mass cultures results in an increase in the number of prechondrogenic condensations and cartilage nodules, whereas overexpression of Prx1b results in a decrease. Prx1a increases the percentage of proliferating cells in micro mass cultures and decreases apoptosis. The Prx1b isoform does not alter proliferation, but it does increase apoptosis, which is opposite of Prx1a. These results suggest that the Prx1a:Prx1b ratio and the alternative splicing mechanism that generates these two isoforms are critical in controlling chondrogenesis. PMID- 15895368 TI - Nephrin expression and three-dimensional morphogenesis of the Xenopus pronephric glomus. AB - Nephrin (NHPS1) encodes a transmembrane protein of approximately 1,200 amino acids that plays a critical role in podocyte slit-diaphragm formation and the development of functional mammalian glomerular filtration barriers. In humans and mice with congenital defects in the nephrin gene, the glomerular filtration barrier is defective and protein leakage into the kidney filtrate causes a life threatening proteinuria. This protein also plays an essential role in the formation of the stellate cells of the Drosophila Malpighian tubules. In this report, the sequence and expression of a Xenopus ortholog of nephrin is described using both conventional and novel three-dimensional (3D) visualization methodologies. Xenopus nephrin encodes a protein of 1,238 amino acids and is expressed at high levels in the forming pronephric kidney glomus, the equivalent of the mammalian glomerulus. Expression commences at stage 25 and is specific to the pronephric glomus up until at least tadpole feeding stages. Two-color fluorescent whole-mount in situ analysis of nephrin expression allowed the 3D shape of the glomus to be imaged and contrasted to the pronephric tubules throughout its morphogenesis. Confocal data processing pipelines were established to generate both volumetric and surface models of the developing pronephros, and a Web-based visualization system was used to generate dynamic and manipulable models of the forming nephric organs. This system allows simple on-line morphometric analysis of the developing pronephric components. As in fish embryos, the glomera first form laterally then migrate medially as the pronephros matures. Unlike in the zebrafish, in Xenopus, this migration stops short of complete fusion of the two glomera at the midline, but a nephrin-positive glomeral nexus does form anteriorly and links the two structures from stage 38 onward. PMID- 15895369 TI - Zic4, a zinc-finger transcription factor, is expressed in the developing mouse nervous system. AB - Zic genes comprise a family of transcription factors, characterized by the presence of a zinc-finger domain containing two cysteines and two histidines (C2 H2). Whereas the embryonic expression patterns of Zic1, 2, 3, and 5 have been described in detail, Zic4 has not yet received close attention. We studied the expression of Zic4 by in situ hybridization during mouse embryogenesis. Zic4 mRNA was first detected at low intensity at embryonic day (E) 9 and, by E10.5, expression was up-regulated in the dorsal midline of the forebrain with a strong, expanded expression domain at the boundary between the diencephalon and telencephalon, the septum, and the lamina terminalis. The choroid plexus of the third ventricle expresses Zic4, as does the dorsal part of the spinal neural tube, excluding the roof plate. The dorsal sclerotome and the dorsomedial lip of the dermomyotome also express Zic4 whereas dorsal root ganglia are negative. At E12.5, Zic4 continues to be expressed in the midline of the forebrain and in the dorsal spinal neural tube. Postnatally, Zic4 is expressed in the granule cells of the postnatal day 2 cerebellum, and in the periventricular thalamus and anterior end of the superior colliculus. We conclude that Zic4 has an expression pattern distinct from, but partly overlapping with, other members of the Zic gene family. PMID- 15895370 TI - Lack of benefit of spinal irradiation in the primary treatment of intracranial germinoma: a multiinstitutional, retrospective review of 180 patients. AB - BACKGROUND: The current study assessed the contribution of spinal irradiation to the treatment outcome of patients with intracranial germinoma. METHODS: Clinical data from 180 patients with intracranial germinoma, who were treated with radiotherapy and/or chemotherapy from 1980 to 2001, were collected from 6 institutions. The patients' median age was 16 years (range, 1-47 yrs), and the male-to-female ratio was 133:47. Pathologic verification was obtained in 88 patients. A solitary tumor was seen in 129 patients, and multifocal or disseminated tumors were detected in 51 patients. The median tumor size was 2.5 cm (range, 0.6-7.0 cm). Local field and/or whole brain irradiation was performed in 114 patients, and craniospinal irradiation was performed in 66 patients. Fifty five patients were treated with chemotherapy. The median follow-up time was 89 months (range, 3-297 mos). RESULTS: Eight-year overall and event-free survival rates were 91% and 89%, respectively. The 8-year recurrence rates at the primary site, intracranial space, and the spinal space were 1%, 6%, and 6%, respectively. Cox regression analysis showed that spinal irradiation (hazard ratio, 1.050; 95% confidence interval [CI], 0.355-3.170) did not contribute to a favorable event free survival. CONCLUSIONS: Spinal irradiation did not contribute to favorable event-free survival in patients with intracranial germinoma. PMID- 15895372 TI - Vascular targeting therapies for treatment of malignant disease. PMID- 15895373 TI - Chondrosarcoma of the scapula: long-term oncologic outcome. AB - BACKGROUND: Chondrosarcoma is the second most common primary sarcoma of bone. It often develops within flat bones, such as the pelvis, ribs, and scapula. In the current study, the authors reviewed the surgical experience and long-term oncologic outcomes of patients with chondrosarcoma arising in the scapula. METHODS: The medical records of 29 consecutive patients with chondrosarcoma of the scapula were reviewed. The patients were treated between 1954 and 1994. All patients had localized disease at the time of presentation. The tumors were classified histologically as Grade 1 (10 patients), Grade 2 (10 patients), Grade 3 (7 patients), dedifferentiated (1 patient), and mesenchymal (1 patient) (using the criteria of Evans et al.). The mean maximal dimension of the tumors was 11 cm. Twenty-five patients underwent limb-sparing surgical resection and 4 patients underwent forequarter amputations. The median follow-up was 13 years (range, 1-35 years). RESULTS: At last follow-up, 22 patients (76%) were free of disease and 7 patients (24%) had died of their disease. Local recurrence occurred in 4 patients at 7 months, 16 months, 40 months, and 43 months, respectively. The local recurrence-free survival rate was 86% at 5 years, 10 years, and 20 years. Disease specific survival was 83% at 5 years, 74% at 10 years, and 74% at 20 years. Patients who had low-grade chondrosarcomas had better survival compared with patients who had high-grade chondrosarcomas (P = 0.07). CONCLUSIONS: Patients who had localized chondrosarcoma of the scapula had a favorable long-term outcome, most likely due to the unique anatomic features that improved the likelihood of achieving wide surgical margins with limb-sparing surgery, despite the frequent presentation of locally advanced disease. PMID- 15895374 TI - Zoledronic acid down-regulates adhesion molecules of bone marrow stromal cells in multiple myeloma: a possible mechanism for its antitumor effect. AB - BACKGROUND: Myeloma plasma cells interact with the bone marrow microenvironment which, in turn, supports their growth and protects them from apoptosis. In vitro studies have demonstrated the antitumor potential of zoledronic acid (ZOL) on myeloma cell lines, but few data are available on its effects on bone marrow stromal cells (BMSCs). The aim of the current study was to evaluate the antiproliferative and apoptotic effect of ZOL on BMSCs, as well as its effect on the expression of adhesion molecules. METHODS: BMSCs, obtained from bone marrow mononucleated cells of 8 patients with multiple myeloma, were treated with increasing concentrations of ZOL for 3 days. Cytotoxic effect was analyzed by 3 (4-5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide; thiazolyl blue (MTT) assay whereas the induction of apoptosis was evaluated by flow cytometric detection of fluorescein isothiocyanate-labeled annexin V, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay, and nuclear changes. Moreover, expression of CD106, CD56, CD50, CD49d, CD44, and CD40 was analyzed by flow cytometry. Data were evaluated by the Friedman test. RESULTS: After 3 days of exposure at concentrations of 10(-4) to 10(-5) M, ZOL induced a decrease in proliferation (P < 0.0001) and an increase in apoptosis (P < 0.002). Analysis of culture supernatants showed that myeloma BMSCs expressed interleukin (IL)-6, negligible levels of tumor necrosis factor-alpha, and no IL-1beta. In vitro exposure to the lowest concentrations of ZOL decreased IL-6 production by BMSCs. Among the adhesion molecules, CD106, CD54, CD49d, and CD40, which were strongly expressed at baseline, showed a statistically significant reduction compared with controls after exposure to ZOL. CONCLUSIONS: ZOL interfered with myeloma BMSCs by reducing proliferation, increasing apoptosis, and modifying the pattern of expression of adhesion molecules, especially those involved in plasma cell binding. These effects on BMSCs might explain the antitumor activity of ZOL. PMID- 15895375 TI - Reclassification of patients with pT3 and pT4 renal cell carcinoma improves prognostic accuracy. AB - BACKGROUND: The significance of adrenal invasion and tumor thrombus in renal cell carcinoma (RCC) has been debated recently. The authors evaluated the associations of direct adrenal invasion, perinephric fat invasion, and tumor thrombus level with outcome to determine whether reclassification would improve the prognostic accuracy of the current primary tumor classification. METHODS: The authors studied 697 patients treated with nephrectomy for pT3 and pT4 RCC between 1970 and 2000. Associations with outcome were evaluated using Cox proportional hazards regression and prognostic accuracy was measured using the c index. RESULTS: Among patients with pT3 RCC, direct adrenal invasion was significantly associated with death from RCC (risk ratio, 2.11; P = 0.004). No significant difference in survival was found between patients with pT4 RCC and pT3 tumors with direct adrenal invasion (P = 0.490). Among patients with pT3b RCC, those with level I III tumor thrombus were significantly more likely to die of RCC compared with patients harboring level 0 tumor thrombus (risk ratio, 1.62; P < 0.001). In addition, patients with fat invasion were more likely to die of RCC compared with pT3 patients without fat invasion (risk ratio, 1.87; P < 0.001). Therefore, patients with pT3 RCC were reclassified into 4 prognostic groups, and this reclassification significantly improved prediction of death from RCC compared with the current classification (c indices of 0.61 vs. 0.55, respectively). CONCLUSIONS: Direct adrenal invasion from RCC should be reclassified as pT4. In addition, the proposed reclassification for patients with pT3 RCC improved prognostic accuracy. PMID- 15895376 TI - Increase in platelet count in older, poor-risk patients with acute myeloid leukemia or myelodysplastic syndrome treated with valproic acid and all-trans retinoic acid. AB - BACKGROUND: The authors investigated the efficacy and safety of the histone deacetylase inhibitors valproic acid (VPA) and all-trans retinoic acid (ATRA) as differentiation agents in a cohort of older, poor-risk patients with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). METHODS: Twenty older patients with recurrent or refractory AML or MDS were treated in a Phase II protocol with sequential VPA and ATRA therapy. VPA was started at a dose of 10 mg/kg per day and then escalated to achieve the serum concentration of 45-100 microg/mL. ATRA was added at 45 mg/square meters (sm) per day when VPA reached the target serum concentration. Only patients treated continuously for > or = 2 months were considered evaluable. RESULTS: Hematologic improvement, according to World Health Organization criteria, was observed in 6 of 20 patients enrolled in the protocol but in 6 of 11 considered evaluable. In five patients, a major platelet response was observed, achieving platelet transfusion independence. Three of these five patients also exhibited a minor erythroid response. A sixth patient showed both a minor erythroid response and a platelet response. The median duration of response was 189 days (range, 63-550 days). No significant reduction in the blast count was observed. Grade 3 neurocortical toxicity was observed in four patients. Severe bone pain was experienced by 4 patients (2 Grade 4 and 2 Grade 3) and was associated with an increase in the peripheral blast cell count. Treatment with ATRA did not modify the response observed with VPA alone. CONCLUSIONS: Differentiation therapy with VPA was of clinical benefit in approximately 30% of elderly patients with AML and MDS of the refractory anemia with excess of blast type with unfavorable prognostic features. A striking platelet transfusion independence lasting several months may be obtained in some patients, reducing the burden of palliative care and improving the quality of life. PMID- 15895377 TI - CpG methylation at promoter site -140 inactivates TGFbeta2 receptor gene in prostate cancer. AB - BACKGROUND: The action of transforming growth factor beta (TGF-beta) is mediated through type 1 (TbetaRI) and type 2 (TbetaRII) receptors. Prostate cancer cells are often resistant to TGF-beta signaling due to loss of TbetaRII expression. The authors of the current study hypothesized that CpG methylation of the TbetaRII promoter at the Sp1 binding site -140 mediates this loss of TbetaRII expression in prostate cancer. METHODS: Sixty-seven prostate cancer (PC) samples, 8 benign prostatic hyperplasia (BPH) samples, and 4 prostate cancer cell lines (DUPro, LNCaP, ND-1 and PC-3) were analyzed for 1) TbetaRII mRNA expression by semiquantitative RT-PCR, 2) TbetaRII protein expression by immunohistochemistry, and 3) TGFbetaRII promoter methylation at CpG site -140 by methylation specific PCR and bisulfite DNA sequencing. Prostate cancer cell lines were treated with the demethylating agent 5aza2'deoxycytidine to determine if TbetaRII gene expression could be increased by blocking promoter methylation. RESULTS: mRNA and protein expression of TbetaRII was lower in the PC samples than in the BPH samples. CpG methylation at site -140 was higher in PC than in BPH (P < 0.01). Promoter methylation was inversely correlated with TbetaRII mRNA expression in the PC and BPH samples (P < 0.0001). PC3, ND1, and DUPro TbetaRII mRNA expression increased following treatment of cells with 5-aza-2'-deoxycytidine. CONCLUSION: CpG methylation of the TbetaRII promoter at CPG site -140 leads to functional loss of the TbetaRII gene in prostate cancer. Treatment with 5-aza-2' deoxycytidine can restore gene expression. The current study results report the first association between prostate cancer and loss of the TGF- beta signaling pathway by TbetaRII DNA promoter methylation. PMID- 15895378 TI - Outcomes and prognostic factors after recurrence in children and adolescents with nonmetastatic rhabdomyosarcoma. AB - BACKGROUND: Although > 90% of children with nonmetastatic rhabdomyosarcoma (RMS) achieve complete remission with current treatment, up to one-third of them experience a recurrence. Survival rates are not always poor in patients who develop recurrences; thus, prognostic factors are needed to tailor salvage treatment. METHODS: The current analysis included 125 children who were affected by localized RMS and were enrolled in 3 consecutive Italian protocols (RMS79, RMS88, and RMS96) who developed recurrences after complete remission. Patient, tumor, and treatment characteristics were studied in univariate and multivariate analyses to determine the independent significance of different factors. RESULTS: The median time from diagnosis to recurrence was 17.8 months. Most patients had local recurrences (72%). The 5-year overall survival (OS) rate was 28.3% +/- 8.7%. Multivariate analysis identified 4 factors that were associated with poor survival: 1) alveolar subtype (relative risk [RR], 2.0), 2) parameningeal or "other" sites (RR, 2.6), 3) systemic recurrence (RR, 3.1), and 4) recurrence on therapy (RR, 2.3). The absence of any of these risk factors identified a "favorable risk" group (12% of patients) with a 5-year OS rate of 71.8% +/- 23.5%. Patients with a single risk factor (32%) had an OS rate of 37.5% +/- 17.2%. Combining patients with 0 or 1 risk factor, the OS rate was 66.5% in the subgroup who had not received radiotherapy compared with an OS rate of 30.3% in the subgroup who had received radiotherapy; this difference was significant (P = 0.03). CONCLUSIONS: The results of the current analysis demonstrated that groups with a different prognosis can be identified among patients with recurrent RMS. Patients with a nonalveolar histology, a primary site other than the parameningeal or "other" sites, local recurrence, and recurrence off therapy had a better prognosis. First-line treatment may have an impact on prognostic variables. In fact, patients who had no or only one risk factor and patients who had tumors with a nonalveolar histology benefited more from salvage therapy if they had not received radiotherapy for their initial treatment. These data may be useful in planning risk-adapted salvage protocols. PMID- 15895379 TI - Controlling protein transport in ultrafiltration using small charged ligands. AB - Previous studies have demonstrated that protein transport during ultrafiltration can be strongly influenced by solution pH and ionic strength. The objective of this study was to examine the possibility of controlling protein transmission using a small, highly charged ligand that selectively binds to the protein of interest. Experiments were performed using bovine serum albumin and the dye Cibacron Blue. Protein sieving data were obtained with essentially neutral and negatively charged versions of a composite regenerated cellulose membrane to examine the effects of electrostatic interactions. The addition of only 1 g/L of Cibacron Blue to an 8 g/L BSA solution reduced the BSA sieving coefficient through the negatively-charged membrane by more than two orders of magnitude, with this effect being largely eliminated at high salt and with the neutral membrane. Protein sieving data were in good agreement with model calculations based on the partitioning of a charged sphere in a charged pore accounting for the change in net protein charge due to ligand binding and the increase in solution ionic strength due to the free ligand in solution. PMID- 15895380 TI - Virus detection using filament-coupled antibodies. AB - Two attractive features of ELISA are the specificity of antibody-antigen recognition and the sensitivity achieved by enzymatic amplification. This report describes the development of a non-enzymatic molecular recognition platform adaptable to point-of-care clinical settings and field detection of biohazardous materials. This filament-antibody recognition assay (FARA) is based on circumferential bands of antibody probes coupled to a 120 microm diameter polyester filament. One advantage of this design is that automated processing is achieved by sequential positioning of filament-coupled probes through a series of 25-60 microL liquid filled microcapillary chambers. This approach was evaluated by testing for the presence of M13KO7 bacterial virus using anti-M13KO7 IgG(1) monoclonal antibody coupled to a filament. Filament motion first positioned the antibodies within a microcapillary tube containing a solution of M13KO7 virus before moving the probes through subsequent chambers, where the filament-coupled probes were washed, exposed to a fluorescently labeled anti-M13K07 antibody, and washed again. Filament fluorescence was then measured using a flatbed microarray scanner. The presence of virus in solution produced a characteristic increase in filament fluorescence only in regions containing coupled antibody probes. Even without the enzymatic amplification of a typical ELISA, the presence of 8.3 x 10(8) virus particles produced a 30-fold increase in fluorescence over an immobilized negative control antibody. In an ELISA comparison study, the filament based approach had a similar lower limit of sensitivity of approximately 1.7 x 10(7) virus particles. This platform may prove attractive for point-of-care settings, the detection of biohazardous materials, or other applications where sensitive, rapid, and automated molecular recognition is desired. PMID- 15895381 TI - Continuous plug-flow bioreactor: experimental testing with Pseudomonas putida culture grown on benzoate. AB - The goals of this work were to test the feasibility of a continuous plug-flow (PF) bioreactor and to compare the growth in the PF bioreactor to that in a batch bioreactor. A culture of Pseudomonas putida was pumped through a tube made of Teflon with varying residence times. The culture was aerated by pumping of air simultaneously with liquid medium to provide air bubbles along the tubular culture. When the residence time in the PF bioreactor was greater than the time needed to reach the stationary phase in batch mode, the maximum biomass density reached in PF mode was the same as the maximum density reached in the batch bioreactor, and benzoate (the only carbon and energy source) was completely consumed. The drawbacks for practical application of PF were found to be fluctuations of cell concentration in the outflow cultural liquid due to cell aggregation, significant cell adhesion to the inner wall of Teflon tubing, and inadequate aeration. PMID- 15895382 TI - Perfusion bioreactor system for human mesenchymal stem cell tissue engineering: dynamic cell seeding and construct development. AB - Human mesenchymal stem cells (hMSCs) have great potential for therapeutic applications. A bioreactor system that supports long-term hMSCs growth and three dimensional (3-D) tissue formation is an important technology for hMSC tissue engineering. A 3-D perfusion bioreactor system was designed using non-woven poly (ethylene terepthalate) (PET) fibrous matrices as scaffolds. The main features of the perfusion bioreactor system are its modular design and integrated seeding operation. Modular design of the bioreactor system allows the growth of multiple engineered tissue constructs and provides flexibility in harvesting the constructs at different time points. In this study, four chambers with three matrices in each were utilized for hMSC construct development. The dynamic depth filtration seeding operation is incorporated in the system by perfusing cell suspensions perpendicularly through the PET matrices, achieving a maximum seeding efficiency of 68%, and the operation effectively reduced the complexity of operation and the risk of contamination. Statistical analyses suggest that the cells are uniformly distributed in the matrices. After seeding, long-term construct cultivation was conducted by perfusing the media around the constructs from both sides of the matrices. Compared to the static cultures, a significantly higher cell density of 4.22 x 10(7) cell/mL was reached over a 40-day culture period. Cellular constructs at different positions in the flow chamber have statistically identical cell densities over the culture period. After expansion, the cells in the construct maintained the potential to differentiate into osteoblastic and adipogenic lineages at high cell density. The perfusion bioreactor system is amenable to multiple tissue engineered construct production, uniform tissue development, and yet is simple to operate and can be scaled up for potential clinical use. The results also demonstrate that the multi-lineage differentiation potential of hMSCs are preserved even after extensive expansion, thus indicating the potential of hMSCs for functional tissue construct development. The system has important applications in stem cell tissue engineering. PMID- 15895383 TI - Diastolic coronary artery compression in a cardiac transplant recipient: treatment with a stent. AB - Myocardial bridges, with resultant systolic compression of the coronary artery, are common inborn anomalies that generally have a benign course. Diastolic compression of the coronary artery, however, is a rare finding that is believed to be an acquired lesion. It can be hypothesized that during diastole, when left ventricular filling occurs, the coronary artery is compressed against epicardial scar tissue or a noncompliant pericardium. This can then lead to diminished intracoronary blood flow. We present a case of functionally significant diastolic coronary artery compression in a cardiac transplant recipient who was successfully treated with intracoronary stent placement. PMID- 15895385 TI - Computations on the A-X transition of isoprene-OH-O2 peroxy radicals. AB - Calculations are carried out on the A state of HO2, CH3O2, and CH3CH2O2 and 10 isomers and conformers of the isoprene-OH-O2 peroxy radicals derived from OH addition to isoprene (2-methyl-1,3-butadiene). In addition to calculating vertical and adiabatic excitation energies, we consider the effect of excitation on molecular structure, and examine the OO stretching frequencies, which are known to be major features in the absorption spectra of the A states of the smaller radicals. The two methods used are the configuration interaction with single excitations (CIS) method and time-dependent density functional theory (TD DFT), both with a range of basis sets up to 6-311++G(2df,2pd). TD-DFT overestimates excitation energies considerably, while CIS tends to underestimate them slightly. TD-DFT does seem to capture the trend in excitation energy vs. size for the smaller peroxy radicals. Conformation and configuration strongly affect the excitation energies of the peroxy radicals from isoprene. CIS calculations indicate that the intramolecular OH--O hydrogen bonds, present in the ground state of some peroxy radicals from isoprene, are weakened or broken in the excited state, while TD-DFT calculations suggest they are retained. PMID- 15895384 TI - Developments in the prediction of type 1 diabetes mellitus, with special reference to insulin autoantibodies. AB - The prodromal phase of type 1 diabetes is characterised by the appearance of multiple islet-cell related autoantibodies (Aab). The major target antigens are islet-cell antigen, glutamic acid decarboxylase (GAD), protein-tyrosine phosphatase-2 (IA-2) and insulin. Insulin autoantibodies (IAA), in contrast to the other autoimmune markers, are the only beta-cell specific antibodies. There is general consensus that the presence of multiple Aab (> or = 3) is associated with a high risk of developing diabetes, where the presence of a single islet cell-related Aab has usually a low predictive value. The most commonly used assay format for the detection of Aab to GAD, IA-2 and insulin is the fluid-phase radiobinding assay. The RBA does not identify or measure Aab, but merely detects its presence. However, on the basis of molecular studies, disease-specific constructs of GAD and IA-2 have been employed leading to somewhat improved sensitivity and specificity of the RBA. Serological studies have shown epitope restriction of IAA that can differentiate diabetes-related from unrelated IAA, but current assays do not distinguish between disease-predictive and non predictive IAA or between IAA and insulin antibodies (IA). More recently, phage display technology has been successful in identifying disease-specific anti idiotopes of insulin. In addition, phage display has facilitated the in vitro production of antibodies with high affinity. Identification of disease-specific anti-idiotopes of insulin should enable the production of a high affinity reagent against the same anti-idiotope. Such a development would form the basis of a disease-specific radioimmunoassay able to identify and measure particular idiotypes, rather than merely detect and titrate IAA. PMID- 15895386 TI - A miniaturized nebulization catheter for improved gene delivery to the mouse lung. AB - BACKGROUND: The available methods for administration of gene delivery systems to the lungs of small animals via nebulization have several drawbacks. These include lack of control over the delivered dose and a negative impact on the stability of the formulation. This paper describes a new nebulization catheter device for the administration of plasmid-based gene delivery systems (polyplexes) as aerosols to the mouse lung in vivo. METHODS: The physical stability of naked pDNA and polyplexes formulated with chitosan oligomers and PEI was examined following nebulization with the catheter device. We also examined the in vitro transfection efficiency of the polyplexes recovered after nebulization. Lung distribution and gene expression after administration of the selected gene delivery systems to the mouse lung were also investigated. RESULTS: In contrast to previously described nebulization methods, the structural integrity of the unprotected naked pDNA was maintained following nebulization by the catheter device, which indicates relatively mild nebulization conditions. In addition, the nebulization procedure did not affect the physical stability of the formulated polyplexes. Small volumes of the pDNA aerosol (10-20 microl) were delivered in a highly controlled and reproducible manner. The aerosol droplet size varied with the molecular weight of the polycations. Aerosol delivery via this method resulted in improved lung distribution of pDNA polyplexes and a six-fold increase in the efficiency of gene delivery in vivo over that seen with the commonly used intratracheal instillation method. CONCLUSION: The use of the nebulization catheter device provides a promising alternative for aerosol gene delivery to the mouse lung. PMID- 15895387 TI - In vivo cutaneous interferon-gamma gene delivery using novel dicationic (gemini) surfactant-plasmid complexes. AB - BACKGROUND: Localized scleroderma (morphea and linear scleroderma) is a connective tissue disease, accompanied by excessive proliferation and deposition of collagen within the skin, inflammation, vasculopathy and a deranged immune system. Interferon gamma (IFNgamma), an inhibitor of collagen synthesis and an immunomodulator, could be a potential therapeutic agent if it could be delivered into or expressed locally in affected skin in a non-invasive manner. In this study, the feasibility of topical delivery of the IFNgamma gene and expression of IFNgamma were investigated in mice. METHODS: Novel dicationic (gemini) surfactant (spacer length n=2-16; alkyl chain m=12 or 16)-DNA complexes were formulated and characterized by circular dichroism and atomic force microscopy to select gemini analogues with the highest transfection efficiency (TE). Transfection and cellular expression of IFNgamma from the bicistronic pGTmCMV.IFN-GFP plasmid were evaluated in PAM 212 keratinocyte culture by ELISA and fluorescence microscopy. Topical delivery of plasmid using liposomal and nanoemulsion systems, based on gemini surfactant 16-3-16, was evaluated in mice by IFNgamma expression analysis. RESULTS: In vitro TE was found to be dependent on the spacer length of the gemini surfactant, with the C3 spacer showing the highest activity (both 12-3-12 and 16 3-16). Both gemini cationic liposomes and gemini nanoemulsion (3x25 microg DNA/animal) produced significantly higher levels of IFNgamma in the skin (359.4 and 607.24 pg/cm2) compared to naked DNA (135.69 pg/cm2) or a liposomal Dc-chol formulation (82.15 pg/cm2). IFNgamma expression in the lymph nodes was higher in the animals treated with gemini liposomes (422.74 pg/animal) compared to the nanoemulsion formulation (131.27 pg/animal) or the Dc-chol formulation (82pg/animal). CONCLUSIONS: The feasibility of topical delivery of pGTmCMV.IFN GFP plasmid in mice using gemini cationic surfactant based delivery systems was demonstrated. IFNgamma expression after treatment with gemini-DNA formulations in the skin was 3-5-fold higher compared to the treatment with naked DNA (p<0.05), and 4-6-fold higher than the Dc-chol-DNA complex, indicating a significant advance in topical DNA delivery across intact skin in vivo. PMID- 15895388 TI - Thg-1 pit gene expression in granule cells of the developing mouse brain and in their synaptic targets, mature Purkinje, and mitral cells. AB - We have studied the expression of Thg-1 pit in developing and adult mouse brain by in situ hybridization analysis. We show that, at day 12.5 of embryo development, Thg-1 pit expression is restricted to the rhombic lip, subventricular neuroepithelium/mantle zone, and lateral ganglionic eminence, namely the embryonic brain areas where granule cell precursors originate. Thereafter, Thg-1 pit expression landmarks both differentiative steps and the mature function of granule/interneuron cells in several brain districts, including cerebellum, basal forebrain, olfactory bulb, and hippocampus. In the adult, Thg-1 pit becomes also activated in mitral cells of olfactory bulb and in Purkinje cells of cerebellum, in concomitance with full development of the synaptic contacts that Purkinje and mitral cells establish with granule cells. We conclude that Thg-1 pit is relevant to specification, proliferation/migration, differentiation, and mature function of granule/interneuron cells in different brain districts, as well as to the function of mature, but not immature, Purkinje cells and mitral cells. PMID- 15895389 TI - GABA receptors and nitric oxide ameliorate constrictive collagen remodeling in hyperhomocysteinemia. AB - Elevated plasma levels of homocysteine (Hcy) are associated with vascular dementias and Alzheimer's disease. The role of Hcy in brain microvascular endothelial cell (MVEC) remodeling is unclear. Hcy competes with muscimol, an gamma-amino butyric acid (GABA)-A receptor agonist. GABA is the primary inhibitory neurotransmitter in the brain. Our hypothesis is that Hcy induces constrictive microvascular remodeling by altering GABA-A/B receptors. MVEC from wild type, matrix metalloproteinase-9 (MMP-9) knockout (-/-), heterozygote cystathionine beta synthase (CBS-/+), and endothelial nitric oxide synthase knockout (eNOS-/-) mouse brains were isolated. The MVEC were incorporated into collagen (3.2 mg/ml) gels and the decrease in collagen gel diameter at 24 h was used as an index of constrictive MVEC remodeling. Gels in the absence or presence of Hcy were incubated with muscimol or baclofen, a GABA-B receptor agonist. The results suggested that Hcy-mediated MVEC collagen gel constriction was ameliorated by muscimol, baclofen, MMP-9, and eNOS gene ablations. There was no effect of anti-alpha 3 integrin. However, Hcy-mediated brain MVEC collagen constriction was abrogated with anti-beta-1 integrin. The co-incubation of Hcy with L-arginine ameliorated the Hcy-mediated collagen gel constriction. The results of this study indicated amelioration of Hcy-induced MVEC collagen gel constriction by induction of nitric oxide through GABA-A and -B receptors. PMID- 15895390 TI - LIGHT regulates CD86 expression on dendritic cells through NF-kappaB, but not JNK/AP-1 signal transduction pathway. AB - The members of the tumor necrosis factor (TNF) family play pivotal roles in the regulation of the immune system. LIGHT is a type II transmembrane protein belonging to the TNF family that was originally identified as a weak inducer of apoptosis. This cytokine has been extensively studied for its role in T cell regulation. Recently, we identified its role in inducing maturation of dendritic cells, such as LIGHT upregulated CD86 expression on dendritic cells in our previous report. However, the signal transduction pathway on this regulation remains unknown. In this study, we found that LIGHT activated NF-kappaB, p44/42 MAPK, but not JNK. LIGHT upregulates CD86 expression on DCs through activation of NF-kappaB, but not p44/42 signal pathway, because inhibition of NF-kappaB activity by its inhibitor could blunt the effect of LIGHT in up-regulation of CD86 expression, but neither inhibitor of p44/42 MAPK nor JNK inhibitor has this effect. Thus we demonstrate that LIGHT regulates CD86 expression through NF kappaB signal transduction pathway but neither p44/42 MAPK nor JNK/AP-1 signaling pathway. We conclude that NF-kappaB signal plays a key role in LIGHT-mediated upregulation of CD86 expression. PMID- 15895391 TI - Histone deacetylase 1 is required for cell cycle exit and differentiation in the zebrafish retina. AB - Histone acetylation is an important epigenetic mechanism for the control of eukaryotic transcription. The histone deacetylase 1 (HDAC1) gene has been implicated in controlling the transcription of core cell cycle regulators, but the in vivo role of HDACs in cell cycle regulation is still poorly understood. Loss of HDAC1 activity causes underproliferation in several contexts during vertebrate development. In contrast, we show here that HDAC1 has the opposite effect in the zebrafish visual system, where loss of HDAC1 activity leads to failure of cells to exit the cell cycle in the retina and in the optic stalk. The effect of HDAC1 on cell cycle exit is cell-autonomous, and loss of HDAC1 in the retina leads to up-regulation of cyclin D and E transcripts. These results demonstrate that the in vivo role of HDAC1 in regulating cell cycle progression is region-specific, as HDAC1 promotes cell cycle exit in the retina but stimulates proliferation in other cellular contexts. PMID- 15895392 TI - Trying to compose the puzzle with all the pieces: epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer. AB - Small molecule epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have shown promising activity in patients with non-small cell lung cancer (NSCLC). Gefitinib has been the first of these drugs to be licensed for third line treatment of advanced NSCLC patients. More recently, erlotinib has been shown to be more effective than placebo in increasing overall survival (OAS) and has been approved for NSCLC patients after failure of chemotherapy. However, a large body of clinical and experimental evidence suggests that the benefit from these drugs is limited to a subgroup of patients. The availability of clinical or molecular criteria for predicting sensitivity to EGFR-TKIs is the most relevant issue for their correct use and for planning future research. Determination of EGFR expression is not sufficient to predict sensitivity to EGFR-TKIs. However, several clinical features (female gender, adenocarcinoma/bronchioloalveolar histotype, never-smoking status, Oriental Asian origin) are associated with major clinical responses. The identification of somatic mutations in the tyrosine kinase domain of the EGFR gene represents the most important molecular marker of sensitivity to EGFR-TKIs. These "activating" mutations can be found in a high proportion of gefitinib- or erlotinib-responding patients. However, clinical effectiveness might not be limited to patients carrying EGFR mutations, in which the objective response is probably the detectable effect of apoptosis induction in cancer cells. In fact, clinical efficacy with gefitinib or erlotinib is also observed in another subgroup of patients, in which a tumor growth delay, determined by a block in cancer cell proliferation, could induce a prolonged and clinically relevant disease stabilization. PMID- 15895393 TI - 24p3 in differentiation of myeloid cells. AB - 24p3 is a secreted lipocalin that has been variously related to apoptosis, proliferation, and the neutrophil lineage of blood cells. We have investigated the expression of 24p3 mRNA and protein in myeloid cell lines induced to differentiate by insulin-like growth factor 1 (IGF-1) and the granulocytic-colony simulating factor (G-CSF). Both these growth factors, which cause myeloid cells to differentiate into granulocytes, induced a marked increase in the expression of both 24p3 protein and mRNA. The mRNA especially appeared early after the cells were induced with either IGF-1 or G-CSF, at a time when the cells were still proliferating and are morphologically undifferentiated. 24p3 can be considered an early marker of granulocytic differentiation. PMID- 15895394 TI - Induction of TGF-beta1 in the trabecular meshwork under cyclic mechanical stress. AB - The pathophysiological mechanisms involved in the failure of the trabecular meshwork (TM) to maintain normal levels of aqueous outflow in glaucoma are not yet understood. Aberrant activation of the transforming growth factor beta-1 (TGF beta1) pathway has been implicated in several degenerative diseases. We investigated the possibility that chronic cyclic mechanical stress that affects the TM might result in increased production of TGF-beta1. Primary cultures of TM cells subjected to cyclic mechanical stress (5% stretching, 1 cycle/sec) demonstrate a significant increase in total and biologically active secreted TGF beta1 that was associated with activation of the TGF-beta1 promoter, measured using a recombinant adenovirus expressing the secreted reporter gene secreted alkaline phosphatase protein (SEAP) under the TGF-beta1 gene promoter (AdTGFbeta1 SEAP). Associated changes in the transcription of MMP-2, TIMP-2, and CTGF were assessed by semiquantitative PCR. Immunohistochemical analysis of TGF-beta1 in organ culture of human eyes revealed a generalized accumulation of this protein in the extracellular matrix (ECM) of the TM, while expression of the TGF-beta1 promoter, analyzed using the LacZ reporter gene, was localized in some specific cells within the outflow pathway. Induction of the TGF-beta1 promoter in organ culture was demonstrated using a novel model for cyclic mechanical stress in human perfused anterior segments infected with AdTGFbeta1-SEAP. Given the relevant physiological and pathophysiological roles of TGF-beta1, its induction after cyclic mechanical stress in the TM supports the hypothesis that this cytokine might play a significant role in the physiology of the TM, and contribute to the pathological changes of this tissue in certain forms of glaucoma. PMID- 15895395 TI - Poly(ADP-ribose)polymerase activity is reduced in circulating mononuclear cells from type 2 diabetic patients. AB - Poly(ADP-ribose)polymerase (PARP-1), a nuclear enzyme activated by DNA strand breaks, is involved in DNA repair, aging, inflammation, and neoplastic transformation. In diabetes, reactive oxygen and nitrogen species occurring in response to hyperglycemia cause DNA damages and PARP-1 activation. Because circulating mononuclear cells (MNCs) are involved in inflammation mechanisms, these cells were chosen as the experimental model to evaluate PARP-1 levels and activity in patients with type 2 diabetes. MNCs were isolated from 25 diabetic patients (18 M, 7 F, age, 63.5 +/- 10.2 years, disease duration 17.7 +/- 8.2 years) and 11 age and sex matched healthy controls. PARP-1 expression and activity were analyzed by semi-quantitative PCR, Western and activity blot, and immunofluorescence microscopy. PARP-1-mRNA expression was increased in MNCs from all diabetic patients versus controls (P < 0.01), whereas PARP-1 content and activity were significantly lower in diabetic patients (P < 0.0001). To verify whether low PARP-1 levels and activity were due to a proteolytic effect of caspase-3 like, the latter activation was measured by a fluorimetric assay. Caspase-3 activity in MNCs was significantly higher in diabetic patients versus control subjects (P < 0.0001). The different PARP-1 behavior in MNCs from patients with type 2 diabetes could therefore be responsible for the abnormal inflammation and infection responses in diabetes. PMID- 15895396 TI - Meeting report on the 12th International Congress of Histochemistry and Cytochemistry (ICHC). AB - The International Congress of Histochemistry and Cytochemistry (ICHC) promoted in San Diego La Jolla (CA, USA), the 12th meeting where researchers of all over the world presented their work and the most innovative methods in histochemical disciplines. A summary of the last meeting is reported. PMID- 15895397 TI - Regulating fragile X gene transcription in the brain and beyond. AB - The past several years have seen remarkable growth in our understanding of the molecular processes underlying fragile X syndrome (FXS). Many studies have provided new insights into the regulation of Fmr1 gene expression and the potential function of its protein product. It is now known that the promoter elements modulating Fmr1 transcription involve a complex array of both cis and trans factors. Moreover, recent studies of epigenetic modification of chromatin have provided novel clues to unlocking the mysteries behind the regulation of Fmr1 expression. Here, we review the latest findings on the regulation of Fmr1 transcription. PMID- 15895398 TI - Oxygen modifies artery differentiation and network morphogenesis in the retinal vasculature. AB - The mechanisms that control differentiation of immature blood vessels into either arteries or veins are not well understood. Because oxygen tension in arteries is higher than in veins, oxygen has the potential to be an instructive signal for artery/vein (AV) differentiation. We test this hypothesis by exposing newborn mice to moderate hypoxia (10% atmospheric oxygen) and studying AV differentiation in the developing retinal vasculature. Forming retinal arteries fail to express the artery-specific markers Delta-like 4 (Dll4) and EphrinB2 during hypoxia. However, other aspects of AV differentiation are retained such as high levels of alpha smooth muscle actin in arterial mural cells and vein-specific expression of the msr/apj gene. The capillary network between arteries and veins is denser, and capillaries expressing the venous marker msr/apj are found in territories normally occupied by arterial capillaries. Thus, it appears that high oxygen in arterial blood is required for arterial expression of Dll4 and EphrinB2, which could be involved in cell-cell repulsion pathways that dictate the normal segregation of arteries and veins. PMID- 15895400 TI - Basement membrane composition in the early mouse embryo day 7. AB - Basement membranes (BM) are specialized structures of the extracellular matrix known to be involved in various early developmental processes. Despite numerous investigations on the localization of BM components, it remains unknown which molecules are expressed in early developmental stages and by which germ layers these proteins are produced. Therefore, we tested for all known laminin chains, nidogens, collagen type IV, and perlecan by means of light microscopic immunostaining and performed in situ reverse transcriptase-polymerase chain reaction to detect the mRNAs specific for laminin alpha1, laminin beta1, the alpha1 chain of collagen type IV, nidogen-2, and perlecan in the early mouse embryo, day 7, in vivo. Only the laminin chains alpha1, beta1, and gamma1 were detected immunohistochemically throughout the entire endodermal and ectodermal BM zones of the embryo proper. The mRNA of laminin alpha1, laminin beta1, collagen type IV, nidogen-2 and perlecan were expressed in the ectoderm-derived mesoderm, in the endoderm as well as in the ectoderm. In contrast, Reichert's membrane was positive for all laminin chains except for the alpha4, alpha5, beta3, and gamma3 chains. Moreover, maternal epithelial as well as mesenchymal cells expressed laminins, nidogen-1 and nidogen-2, collagen type IV, and perlecan. In conclusion, laminin-1 might be the only laminin isoform in the early mouse embryo that, together with the other main BM components, nidogens, collagen type IV, and perlecan, is synthesized by all three germ layers. PMID- 15895401 TI - Prevalence of and risk factors for nonalcoholic fatty liver disease: the Dionysos nutrition and liver study. AB - The prevalence of and the risk factors for fatty liver have not undergone a formal evaluation in a representative sample of the general population. We therefore performed a cross-sectional study in the town of Campogalliano (Modena, Italy), within the context of the Dionysos Project. Of 5,780 eligible persons aged 18 to 75 years, 3,345 (58%) agreed to participate in the study. Subjects with suspected liver disease (SLD), defined on the basis of elevated serum alanine aminotransferase (ALT) and gamma-glutamyl-transferase (GGT) activity, hepatitis B surface antigen (HBsAg), or hepatitis C virus (HCV)-RNA positivity, were matched with randomly selected subjects of the same age and sex without SLD. A total of 311 subjects with and 287 without SLD underwent a detailed clinical, laboratory, and anthropometrical evaluation. Fatty liver was diagnosed by ultrasonography, and alcohol intake was assessed by using a 7-day diary. Multinomial logistic regression was used to detect risk factors for normal liver versus nonalcoholic fatty liver disease (NAFLD) and for alcoholic fatty liver (AFLD) versus NAFLD. The prevalence of NAFLD was similar in subjects with and without SLD (25 vs. 20%, P = .203). At multivariable analysis, normal liver was more likely than NAFLD in older subjects and less likely in the presence of obesity, hyperglycemia, hyperinsulinemia, hypertriglyceridemia, and systolic hypertension; AFLD was more likely than NAFLD in older subjects, males, and in the presence of elevated GGT and hypertriglyceridemia, and less likely in the presence of obesity and hyperglycemia. In conclusion, NAFLD is highly prevalent in the general population, is not associated with SLD, but is associated with many features of the metabolic syndrome. PMID- 15895402 TI - Angiographic predictors of femoral access site complications: implication for planned percutaneous coronary intervention. AB - This study examined the relationship between the femoral arteriotomy location and the risk of femoral access site complications after diagnostic and interventional cardiac catheterization procedures. One of the most common complication of cardiac catheterization and percutaneous coronary intervention (PCI) involves the vascular access site. The femoral approach is the most frequent site of vascular access during invasive cardiac procedures. This approach is associated with vascular complications, such as retroperitoneal bleeding, which can be life threatening. If angiographic predictors of retroperitoneal bleeding can be identified, this complication could be avoided. A prospective cohort of 33 patients with femoral access site complications was subgrouped based on the angiographic arteriotomy site. Concurrent patients without complications were randomly selected to form a control group. Study and control patients were compared on presenting risk factors and outcomes. Logistic regression analysis was used to identify independent predictors for femoral access site complications. Arteriotomy location above the most inferior border of the inferior epigastric artery in patients undergoing PCI was associated with 100% of all retroperitoneal bleeds (P < 0.001). Low, high middle, and high femoral arteriotomy sites were associated with 71% of all vascular access complications. The combination of these locations for the femoral arteriotomy was an independent predictor of adverse vascular access site complications beyond traditional risk factors (odds ratio = 28.7; CI = 6.73-122.40; P < 0.0001). Vascular complications occurred more frequently in patients who were of older age (72 vs. 66 years; P < 0.001). The location of the femoral arteriotomy site assessed by a femoral angiogram is predictive of life-threatening complications. Patients undergoing PCI with an arteriotomy above the most inferior border of the inferior epigastric artery are at an increased risk for retroperitoneal bleeding. This complication may be avoided by risk-stratifying patients prior to intervention with a femoral angiogram. PMID- 15895403 TI - Percutaneous occlusion of a pulmonary aneurysm causing hemoptysis in a patient with pulmonary atresia and aortopulmonary collaterals. AB - A 28-year-old male was referred for cardiac catheterization because of recurrent severe hemoptysis necessitating resuscitation and subsequently preventing weaning from ventilation. He had a history of pulmonary atresia, ventricular septal defect, overriding aorta with right-sided aortic arch diagnosed at birth. Eisenmenger's syndrome ensued and he was not felt to be suitable for corrective cardiac surgery. He had multiple major aortopulmonary collateral vessels to both lungs with a large aneurysm in an artery to the right lower lobe, which was suspected to be the source of his bleeding. Occlusion of this aneurysm was achieved percutaneously using an Amplatzer septal occluder device. He had no further bleeding and was successfully weaned from ventilation. Six months later, he has recovered to his functional baseline and has not had any further episodes of hemoptysis. PMID- 15895404 TI - Transradial stenting of the iliac artery: a case report. AB - In this case report, stenting of the common iliac artery via the radial access route with 6 Fr equipment, including an extra long multipurpose catheter, is presented. Radial access avoids bleeding and ischemia that may complicate retrograde or crossover access to the iliac artery. PMID- 15895405 TI - Comparative analysis of the natriuretic peptide precursor gene cluster in vertebrates reveals loss of ANF and retention of CNP-3 in chicken. AB - We identified and characterized the chicken natriuretic peptide precursor gene cluster and found its organization to be highly conserved compared with the mammalian Nppb-Nppa cluster. However, phylogenetic analysis indicated that the putative chicken natriuretic peptide precursor genes are the homologues of CNP-3 and Nppb, respectively. Comparative expression analysis revealed that, in human, mouse, and rat hearts, Nppb is a novel marker for the differentiating working myocardium. Its expression pattern is strikingly similar to that of Nppa before birth, and diverges only after birth. In contrast, whereas the chicken Nppb gene expression profile resembled that of mammalian Nppb, the CNP-3 gene showed very limited expression in the heart, not resembling the pattern of either Nppa or Nppb. These results show that, in chicken, the Nppa gene has been lost from the natriuretic peptide precursor gene cluster, whereas the CNP-3 gene has been retained. PMID- 15895406 TI - Left ventricular unloading and concomitant total cardiac output increase by the use of percutaneous Impella Recover LP 2.5 assist device during high-risk coronary intervention. AB - A number of techniques have been proposed for circulatory support during high risk percutaneous coronary interventions (PCI), but no single approach has achieved wide acceptance so far. We report on a patient with severe left ventricular (LV) impairment who underwent a PCI with the use of a new left ventricular assist device, the Impella Recover LP 2.5 system. The effects on global cardiac output were determined by thermodilution (TD) and LV pressure volume loops obtained by conductance catheter. The activation of the pump resulted in a rapid and sustained unloading effect of the LV. At the same time, the continuous expulsion of blood into ascending aorta throughout the cardiac cycle produced by the pump resulted in an increase of systemic overall CO, measured by the TD technique, of 1.43 L/min. The procedure was uncomplicated and the patient remained uneventful at follow-up. Our single experience gives new input for future trials to assess the effect of the Impella Recover LP 2.5 assist device on outcome in this subset of patients. PMID- 15895407 TI - Transgenic zebrafish that express tyrosine hydroxylase promoter in inner retinal cells. AB - We have generated a transgenic zebrafish line [Tg(Th:GFP)] that expresses green fluorescence proteins (GFP) driven by rat tyrosine hydroxylase (TH) promoter. In zebrafish, the transgene was expressed as early as 16 hr postfertilization (hpf). The first transgene expression was detected in the midbrain. Within a few hours of development, the expression spread to the forebrain and hindbrain. In the retina, the first transgene expression was detected at approximately 40 hpf, at which time a single GFP-positive cell was seen in the ventral-nasal patch of the retina. In late development, GFP spread across the inner retina. GFP was found in retinal cells that expressed TH or phenylethanolamine N-methyl-transferase (PNMT), the first and last enzymes for synthesis of catecholamine, respectively. This suggests that the transgene is expressed in catecholaminergic neurons. Of interest, GFP was also detected in some retinal cells that release gamma aminobutyric acid. These latter data suggest that the transgene may also be expressed in noncatecholaminergic cells. PMID- 15895408 TI - Optimizing vascular access: routine femoral angiography keeps the vascular complication away. PMID- 15895409 TI - Analysis of gene expression patterns in the developing chick liver. AB - The chick embryo has been used widely for studying liver development. However, in the past 30 years, the usage has decreased markedly due to lack of appropriate marker genes for differentiation in the developing chick liver. To use the chick embryo for analyzing the molecular mechanism of liver development, we surveyed marker genes in the developing chick liver by examining the expression pattern of genes that are well-characterized in the developing mammalian liver. By whole mount in situ hybridization, Fibrinogen-gamma (FIB) expression was first detected at stage 12, specifically in the anterior intestinal portal, and its liver specific expression persisted in the later stages. Albumin (ALB) expression was first detected at stage 30, when the liver starts maturing. Cytokeratin 19 (CK19) was first detected at stage 37 in the ductal plate of the liver, and its expression continued in the intrahepatic bile ducts derived from the ductal plate. Hex, a transcription factor, is an additional marker of bile duct differentiation. Hence, FIB, ALB, and CK19 expression can be used to trace hepatic induction, maturation, and bile duct differentiation, respectively. PMID- 15895410 TI - Expression pattern of four membrane-type matrix metalloproteinases in the normal and diseased mouse mammary gland. AB - Both mammary gland development and mammary carcinogenesis involve extensive remodeling of the mammary gland extracellular matrix. The expression of four membrane-type matrix metalloproteinases (MT-MMPs) with matrix remodeling potential in development and tumorigenesis was evaluated by in-situ hybridization on mouse mammary gland sections. MT1-MMP and MT3-MMP were found in the mammary stroma mainly around epithelial structures in both developing and mature mammary gland. In contrast, MT2-MMP was found exclusively in the mammary epithelium. Lactating gland expressed none of the examined MT-MMPs. Mammary gland tumors expressed MT1-MMP, MT2-MMP, and MT3-MMP while MT4-MMP was not expressed in any developmental or cancerous stage analyzed here. Our results suggest that MT1-MMP, MT2-MMP, and MT3-MMP may be involved in remodeling of both the normal and diseased mammary gland either directly or indirectly by activation of other MMPs. PMID- 15895411 TI - cAMP induced modifications of HOX D gene expression in prostate cells allow the identification of a chromosomal area involved in vivo with neuroendocrine differentiation of human advanced prostate cancers. AB - The acquisition of epithelial-neuroendocrine differentiation (ND) is a peculiarity of human advanced, androgen-independent, prostate cancers. The HOX genes are a network of transcription factors controlling embryonal development and playing an important role in crucial adult eukaryotic cell functions. The molecular organization of this 39-gene network is unique in the genome and probably acts by regulating phenotype cell identity. The expression patterns of the HOX gene network in human prostate cell phenotypes, representing different stages of prostate physiology and prostate cancer progression, make it possible to discriminate between different human prostate cell lines and to identify loci and paralogous groups harboring the HOX genes mostly involved in prostate organogenesis and cancerogenesis. Exposure of prostate epithelial phenotypes to cAMP alters the expression of lumbo-sacral HOX D genes located on the chromosomal region 2q31-33 where the cAMP effector genes CREB1, CREB2, and cAMP-GEFII are present. Interestingly, this same chromosomal area harbors: (i) a global cis regulatory DNA control region able to coordinate the expression of HOX D and contiguous phylogenetically unrelated genes; (ii) a prostate specific ncRNA gene associated with high-risk prostate cancer (PCGEM1); (iii) a series of neurogenic related genes involved with epithelial-neuronal cell conversion. We report the expression of neurexin 1, Neuro D1, dlx1, and dlx2 in untreated and cAMP treated epithelial prostate cells. The in vivo expression of Neuro D1 in human advanced prostate cancers correlate with the state of tumor differentiation as measured by Gleason score. Thus, we suggest that the chromosomal area 2q 31-33 might be involved in the epithelial-ND characteristic of human advanced prostate cancers. PMID- 15895412 TI - Percutaneous implantation of an aortic valve prosthesis. AB - Recently, percutaneous aortic valve implantation has become an alternative technique to surgical valve replacement in patient at high risk for surgery. Our animal experimentation evaluated the technical feasibility of aortic valve replacement using a bovine pericardium valve sutured on a self-expandable stent in a sheep model. Precise implantation with satisfactory attachment on the adjacent tissues and absence of migration was obtain in 8 out of 14 animals. This study confirmed the feasibility of the endovascular implantation of a pericardium valve sutured on a self-expandable stent in a sheep model. PMID- 15895414 TI - Who let the bugs in? PMID- 15895415 TI - Developmental regulation and expression of the zebrafish connexin43 gene. AB - We cloned and sequenced the zebrafish (Danio rerio) connexin43 (Cx43alpha1) gene. The predicted protein sequence shows a high degree of sequence conservation. Transcript analyses revealed multiple transcription start sites and a potential alternative transcript encoding a N-terminally truncated Cx43alpha1 protein. Maternal Cx43alpha1 transcripts were detected, with zygotic expression initiated before gastrulation. In situ hybridization revealed many Cx43alpha1 expression domains, including the notochord and brain, heart and vasculature, many resembling patterns seen in mammalian embryos. Of interest, a reporter construct under control of the mouse Cx43alpha1 promoter was observed to drive green fluorescent protein expression in zebrafish embryos in domains mimicking the native Cx43alpha1 expression pattern in fish and mice. Sequence comparison between the mouse and zebrafish Cx43alpha1 promoter sequences showed the conservation of several transcription factor motifs, which otherwise shared little overall sequence homology. The conservation of protein sequence and developmental gene regulation would suggest that Cx43alpha1 gap junctions are likely to have conserved roles in vertebrate embryonic development. PMID- 15895416 TI - Cloning and functional characterization of a novel connexin expressed in somites of Xenopus laevis. AB - Connexin-containing gap junctions play an essential role in vertebrate development. More than 20 connexin isoforms have been identified in mammals. However, the number identified in Xenopus trails with only six isoforms described. Here, identification of a new connexin isoform from Xenopus laevis is described. Connexin40.4 was found by screening expressed sequence tag databases and carrying out polymerase chain reaction on genomic DNA. This new connexin has limited amino acid identity with mammalian (<50%) connexins, but conservation is higher (approximately 62%) with fish. During Xenopus laevis development, connexin40.4 was first expressed after the mid-blastula transition. There was prominent expression in the presomitic paraxial mesoderm and later in the developing somites. In adult frogs, expression was detected in kidney and stomach as well as in brain, heart, and skeletal muscle. Ectopic expression of connexin40.4 in HEK293 cells, resulted in formation of gap junction like structures at the cell interfaces. Similar ectopic expression in neural N2A cells resulted in functional electrical coupling, displaying mild, asymmetric voltage dependence. We thus cloned a novel connexin from Xenopus laevis, strongly expressed in developing somites, with no apparent orthologue in mammals. PMID- 15895417 TI - Cx31 and Cx43 double-deficient mice reveal independent functions in murine placental and skin development. AB - The overlapping expression of gap junctional connexins in tissues has indicated that the channels may compensate for each other. During development, Cx31 and Cx43 are coexpressed in preimplantation embryos, in the spongiotrophoblast of the placenta and in the epidermis. This study shows that Cx31/Cx43 double-deficient mice exhibit the known phenotypes of the single-knockout strains but no combined effects. Thus, Cx43, coexpressed with Cx31 at midgestation in the spongiotrophoblast of the placenta, cannot be responsible for a partial rescue of the lethal Cx31 knockout phenotype, as assumed before (Plum et al. [2001] Dev Biol 231:334-337). It follows that both connexins have unique functions in placental development. Despite an altered expression of other epidermal connexin mRNAs, epidermal differentiation and physiology was unaltered by the absence of Cx31 and Cx43. Therefore, in epidermal and preimplantation development, gap junctional communication can probably be compensated by other isoforms coexpressed with Cx31 and Cx43. PMID- 15895418 TI - Bioarchaeological analysis of cultural transition in the southern Levant using dental nonmetric traits. AB - To many Near Eastern archaeologists, the Late Bronze Age-Early Iron Age transition in the southern Levant indicates the emergence of a new ethnicity. The question remains, however, whether changes in the material culture are the result of an invasion of foreigners, or instead arose from shifting cultural and technical practices by indigenous peoples. This study utilized dental morphological traits to assess phenetic relationships between the Late Bronze Age site of Dothan (1500-1100 BC) and the Iron Age II site of Lachish (Tell ed Duweir, 701 BC). Information on 30 dental crown and root traits was collected for 4,412 teeth, representing 392 individuals from Lachish and a minimum of 121 individuals from Dothan, using the Arizona State University Dental Anthropology System. Seventeen traits from Dothan and Lachish were compared with dentitions from a Byzantine Jerusalem monastery, Iron Age Italy, a Natufian group (early agrarians from the Levant), and a Middle Kingdom Egyptian site using C.A.B. Smith's mean measure of divergence statistic. The findings suggest that there are more similarities between Dothan and Lachish than either of them and other sites. This analysis indicates that the material culture changes were not the result of a foreign invasion. Rather, the Iron Age people of the southern Levant were related to their Bronze Age predecessors. PMID- 15895419 TI - Body proportions in ancient Andeans from high and low altitudes. AB - Living human populations from high altitudes in the Andes exhibit relatively short limbs compared with neighboring groups from lower elevations as adaptations to cold climates characteristic of high-altitude environments. This study compares relative limb lengths and proportions in pre-Contact human skeletons from different altitudes to test whether ecogeographic variation also existed in Andean prehistory. Maximum lengths of the humerus, radius, femur, and tibia, and femoral head breadth are measured in sex-specific groups of adult human skeletons (N = 346) from the central (n = 80) and the south-central (n = 123) Andean coasts, the Atacama Desert at 2,500 m (n = 102), and the southern Peruvian highlands at 2,000-3,800 m (n = 41). To test whether limb lengths vary with altitude, comparisons are made of intralimb proportions, limb lengths against body mass estimates derived from published equations, limb lengths against the geometric mean of all measurements, and principal component analysis. Intralimb proportions do not statistically differ between coastal groups and those from the Atacama Desert, whereas intralimb proportions are significantly shorter in the Peruvian highland sample. Overall body size and limb lengths relative to body size vary along an altitudinal gradient, with larger individuals from coastal environments and smaller individuals with relatively longer limbs for their size from higher elevations. Ecogeographic variation in relation to climate explains the variation in intralimb proportions, and dietary variation may explain the altitudinal cline in body size and limb lengths relative to body size. The potential effects of gene flow on variation in body proportions in Andean prehistory are also explored. PMID- 15895420 TI - Activity-related skeletal change in medieval humeri: cross-sectional and architectural alterations. AB - This paper examines humeral cross-sectional properties in two different samples of later medieval date: a group of blade-injured males from the sites of Towton, North Yorkshire, and Fishergate in the City of York, England, and a comparative group of nonblade-injured males also from the site of Fishergate in York. CT image slices were taken of the humeral shaft at 20%, 35%, 50%, 65%, and 80% from the distal end to investigate population differences in levels and patterns of mechanical loading. Bilateral asymmetry is investigated and comparisons are made with different populations of varying activity levels. Architectural changes such as humeral torsion are also investigated to determine the relationship between architectural changes and biomechanical efficiency. Results show significant differences in diaphyseal robusticity between the Towton sample and the comparative population, as well as significant differences in diaphyseal shape both between limbs within the Towton sample and between blade-injured samples. Population differences were also identified in the level of bilateral asymmetry, further demonstrating the differences in movement and activity patterns both between and within samples. These variations may relate to distinctive, more strenuous weapon use and differences in strenuous movement patterns in the two groups. PMID- 15895421 TI - Effect of repetitive TMS and fluoxetine on cognitive function in patients with Parkinson's disease and concurrent depression. AB - Previous studies show that cognitive functions are more impaired in patients with Parkinson's disease (PD) and depression than in nondepressed PD patients. We compared the cognitive effects of two types of antidepressant treatments in PD patients: fluoxetine (20 mg/day) versus repetitive transcranial magnetic stimulation (rTMS, 15 Hz, 110% above motor threshold, 10 daily sessions) of the left dorsolateral prefrontal cortex. Twenty-five patients with PD and depression were randomly assigned either to Group 1 (active rTMS and placebo medication) or to Group 2 (sham rTMS and fluoxetine). A neuropsychological battery was assessed by a rater blind to treatment arm at baseline and 2 and 8 weeks after treatment. Patients in both groups had a significant improvement of Stroop (colored words and interference card) and Hooper and Wisconsin (perseverative errors) test performances after both treatments. Furthermore, there were no adverse effects after either rTMS or fluoxetine in any neuropsychological test of the cognitive test battery. The results show that rTMS could improve some aspects of cognition in PD patients similar to that of fluoxetine. The mechanisms for this cognitive improvement are unclear, but it is in the context of mood improvement. PMID- 15895422 TI - Alpha-synuclein and Parkinson's disease: implications from the screening of more than 1,900 patients. AB - Data on the frequency of alpha-synuclein mutations in Parkinson's disease (PD) are limited. Screening the entire coding region in 1,921 PD patients with denaturing high performance liquid chromatography and subsequent sequencing we only detected silent mutations (g.2654A>G, g.10151G>A, and g.15986A>T) and the c.209G>A substitution corresponding to the p.A53T mutation. These results demonstrate that mutations in the alpha-synuclein gene are rare and suggest that other factors contribute to alpha-synuclein aggregation in the majority of PD patients. PMID- 15895423 TI - Speech-activated myoclonus: an uncommon form of action myoclonus. AB - We describe an unusual form of facial myoclonus activated by speech in 3 patients with different underlying neurological diseases and present the electrophysiological investigations and results of structural and functional imaging. In 1 of 2 patients in whom jerk-locked electroencephalogram (EEG) back averaging was done, a cortical potential clearly preceded the facial jerks. In the second patient, a cortical potential preceding the jerk was not certain. In the third patient, the resting EEG contained outbursts of symmetric, slower frequencies of indeterminate significance. An epileptiform disorder was suspected in this patient. PMID- 15895424 TI - Investigation into the correlation between sensation and leg movement in restless legs syndrome. AB - We evaluated rest effects on restless legs syndrome (RLS) sensory and motor symptoms. During two 60-minute Suggested Immobilization Tests (SIT) subject's signals of RLS leg sensations and periodic leg movements while awake (PLMW) were recorded. Sensations, PLMW, sensations preceding or after PLMW, sensations occurring without following PLMW, and PLMW occurring without preceding sensation were determined. The RLS patients were divided into equal-sized high and low PLMW groups for further analysis. Data from 46 subjects (28 RLS and 18 controls) revealed sensations increased linearly with rest in RLS patients and controls. Movement rate increased linearly with rest for controls but increased rapidly for the first 45 minutes for all RLS patients. PLMW/hour increased with further rest for low but not high PLMW patients. Sensations followed by PLMW and PLMW without preceding sensations followed similar patterns. Sensations without subsequent PLMW increased dramatically in the last 15 minutes of the SITs. Whereas both sensory and motor signs of RLS increase with rest, there is minimal increase for controls. Patients with higher but not lower PLMW rates reached a ceiling for PLMW after 35 to 40 minutes. The temporal dissociation between sensory and motor events supports viewing these motor and sensory events as separate but loosely linked manifestations of RLS. PMID- 15895425 TI - Coexistence of movement disorders and epilepsia partialis continua as the initial signs in probable Creutzfeldt-Jakob disease. AB - Movement disorders and epilepsy rarely occur in the early stage of Creutzfeldt Jakob disease (CJD) but have not been reported concurrently. We report on a 47 year-old patient with probable CJD who presented with generalized chorea and focal dystonia with myoclonic jerks on the right hand. Myoclonic jerks progressed to epilepsia partialis continua within 5 days of admission to the hospital. The diagnosis of our patient was compatible with probable CJD on the basis of clinical course, electroencephalogram, and diffusion-weighted magnetic resonance imaging findings, and presence of 14-3-3 protein in cerebrospinal fluid. To our knowledge, this is the first report of a case developing both movement disorders and epilepsia partialis continua in the early stage of the disease. PMID- 15895426 TI - Stimulation of the globus pallidus internus for childhood-onset dystonia. AB - We report the results of deep brain stimulation (DBS) of the globus pallidus internus (GPi) in 12 patients with childhood-onset generalized dystonia refractory to medication, including 3 patients with status dystonicus. There were 8 patients who had DYT1-negative primary dystonia, 1 had DYT1-positive dystonia, and 3 had symptomatic dystonia. Stimulation was effective in all but 1 patient. Dystonic postures and movements of the axis and limbs responded to DBS to a greater extent than oromandibular dystonia and fixed dystonic postures. These findings provide further evidence that pallidal stimulation is an effective treatment for intractable childhood-onset dystonia, including status dystonicus, and together with previous findings, suggest that it should be considered the treatment of choice for these conditions. PMID- 15895427 TI - Purification and characterization of mouse fetal liver epithelial cells with high in vivo repopulation capacity. AB - Epithelial cells in embryonic day (ED) 12.5 murine fetal liver were separated from hematopoietic cell populations using fluorescence-activated cell sorting (FACS) and were characterized by immunocytochemistry using a broad set of antibodies specific for epithelial cells (alpha-fetoprotein [AFP], albumin [ALB], pancytokeratin [PanCK], Liv2, E-cadherin, Dlk), hematopoietic/endothelial cells (Ter119, CD45, CD31), and stem/progenitor cells (c-Kit, CD34, Sca-1). AFP(+)/ALB(+) cells represented approximately 2.5% of total cells and were positive for the epithelial-specific surface markers Liv2, E-cadherin, and Dlk, but were clearly separated and distinct from hematopoietic cells (Ter119(+)/CD45(+)). Fetal liver epithelial cells (AFP(+)/E-cadherin(+)) were Sca 1(+) but showed no expression of hematopoietic stem cell markers c-Kit and CD34. These cells were enriched by FACS sorting for E-cadherin to a purity of 95% as defined by co-expression of AFP and PanCK. Purified fetal liver epithelial cells formed clusters in cell culture and differentiated along the hepatocytic lineage in the presence of dexamethasone, expressing glucose-6-phosphatase (G6P) and tyrosine amino transferase. Wild-type ED12.5 murine fetal liver cells were transplanted into adult dipeptidyl peptidase IV knockout mice and differentiated into mature hepatocytes expressing ALB, G6P, and glycogen, indicating normal biochemical function. Transplanted cells became fully incorporated into the hepatic parenchymal cords and showed up to 80% liver repopulation at 2 to 6 months after cell transplantation. In conclusion, we isolated and highly purified a population of epithelial cells from the ED12.5 mouse fetal liver that are clearly separate from hematopoietic cells and differentiate into mature, functional hepatocytes in vivo with the capacity for efficient liver repopulation. Supplementary material for this article can be found on the HEPATOLOGY website (http://www.interscience.wiley.com/jpages/0270 9139/suppmat/index.html). PMID- 15895428 TI - Response of anterior temporal cortex to syntactic and prosodic manipulations during sentence processing. AB - Previous research has implicated a portion of the anterior temporal cortex in sentence-level processing. This region activates more to sentences than to word lists, sentences in an unfamiliar language, and environmental sound sequences. The current study sought to identify the relative contributions of syntactic and prosodic processing to anterior temporal activation. We presented auditory stimuli where the presence of prosodic and syntactic structure was independently manipulated during functional magnetic resonance imaging (fMRI). Three "structural" conditions included normal sentences, sentences with scrambled word order, and lists of content words. These three classes of stimuli were presented either with sentence prosody or with flat supra-lexical (list-like) prosody. Sentence stimuli activated a portion of the left anterior temporal cortex in the superior temporal sulcus (STS) and extending into the middle temporal gyrus, independent of prosody, and to a greater extent than any of the other conditions. An interaction between the structural conditions and prosodic conditions was seen in a more dorsal region of the anterior temporal lobe bilaterally along the superior temporal gyrus (STG). A post-hoc analysis revealed that this region responded either to syntactically structured stimuli or to nonstructured stimuli with sentence-like prosody. The results suggest a parcellation of anterior temporal cortex into 1) an STG region that is sensitive both to the presence of syntactic information and is modulated by prosodic manipulations (in nonsyntactic stimuli); and 2) a more inferior left STS/MTG region that is more selective for syntactic structure. PMID- 15895429 TI - Ethnic expansions and between-group differences in children's health: a case study from the Rukwa Valley, Tanzania. AB - The Sukuma ethnic group appears to be expanding across Tanzania at a rate far greater than other ethnic groups in the area. In this paper, the household-level dynamics that may be fueling this expansion are explored by comparing measures of infant mortality and child health with another Tanzanian ethnic group, the Pimbwe. Consistent with expectations, the Sukuma appear to have comparable levels of fertility but lower child mortality. As predicted, compared to the Pimbwe, Sukuma children are also heavier and taller for their age, suggesting better nutritional status. Four hypotheses about why the Sukuma are so successful in this area are addressed. Surprisingly, the results show that household food security and wealth are not related to children's nutritional status, nor can maternal effects account for the observed health differences. Several lines of evidence suggest that different patterns of infant feeding practices may underlie the differences in children's nutritional status. PMID- 15895430 TI - Sexual dimorphism in the Japanese cranial base: a Fourier-wavelet representation. AB - An approach, computational shape analysis (CSA), is presented here which utilizes a Fourier-wavelet representation to numerically describe shape features of biological forms. Two elements are involved: 1) elliptical Fourier functions (EFFs), to provide estimates of global aspects, and 2) continuous wavelet transforms (CWTs) to generate an objective estimate of localized features. EFFs are computed, using a set of pseudohomologous points, to create a precise analog of the boundary. This computed contour is then normalized by scaling and rotated in two-dimensional space to insure a representation that is invariant with respect to starting point, size, and orientation. The predicted point coordinates derived from the EFFs are submitted to CWT for further processing. Wavelet coefficients are then computed to identify localized features, localization being a subjective process with EFFs. The advantage of wavelets is that they eliminate the inevitable subjectivity inherent in the choice of measurements. To test the usefulness of CSA, a sample of cranial base (CB) lateral radiographic outlines was available. Five archaeological periods, Yayoi, Kofun, Kamakura, Edo, and Modern, were utilized (n = 297). Statistically significant differences in sex and archaeological age were found. Although archaeological age differences were present, they were small and random in character, suggesting stability in the CB structures. In contrast, sexually dimorphic differences were present in every group from the Yayoi to the Modern period. This presence of sexually dimorphic differences in shape was consistent with earlier studies of M. nemestrina, G. gorilla, and P. troglodytes. Consequently, it is suggested that the pattern of sexual dimorphism documented in the Japanese CB is a primate pattern with an ancient evolutionary history. The results demonstrate, both visually and numerically, that CSA is a powerful approach for describing both global and localized features of craniofacial structures such as the CB. PMID- 15895431 TI - A new set of amino acid descriptors and its application in peptide QSARs. AB - In this work, a new set of amino acid descriptors, i.e., VHSE (principal components score Vectors of Hydrophobic, Steric, and Electronic properties), is derived from principal components analysis (PCA) on independent families of 18 hydrophobic properties, 17 steric properties, and 15 electronic properties, respectively, which are included in total 50 physicochemical variables of 20 coded amino acids. Using the stepwise multiple regression (SMR) method combined with partial least squares (PLS), the VHSE scales are then applied to QSAR studies of bitter-tasting dipeptides (BTD), angiotensin-converting enzyme (ACE) inhibitors, and bradykinin-potentiating pentapeptides (BPP). To validate the predictive power of resulting models, external validation are also performed. A comparison of the results to those obtained with z scores and other two dimensional (2D) or three-dimensional(3D) descriptors shows that the VHSE scales are comparable for parameterizing the structural variability of the peptide series. PMID- 15895432 TI - Mesiodistal tooth crown dimensions of the primary dentition: a worldwide survey. AB - This analysis reports on a spatial-temporal survey of published studies of primary tooth crown dimensions in humans (80 samples). Mesiodistal data are analyzed for the 10 tooth crown dimensions. The purpose was to evaluate the numerous case reports (descriptive analyses of single samples) in the literature in order to assess patterning of variation 1) in tooth size, 2) among tooth types, 3) across sexes, 4) with space (historical affinity), and 5) with time. Sexual dimorphism is low in the primary dentition, averaging 2% across all 10 tooth types. All size distributions of the samples are positively skewed because of megadont native Australians. Europeans, who are most frequently represented in the literature, have the smallest tooth crowns of any continental grouping assessed. The method by Darroch Mosimann ([1985] Biometrika 72:241-252) of reducing size effects was used, basically standardizing the data variable-wise, and then ordinating groups on their factor scores. Principal components analysis produced just two canonical axes: overall size (68%) and a front-back (i1-i2-c vs. m1-m2) polarity (11%), based on the intergroup (not ontogenetic) covariance matrix. This second component discriminates between groups with relatively large anterior teeth (Europeans) and those where relatively more tooth substance is apportioned to the molars (Africans and Asians). Size differences predominate over shape between sexes from the same groups. Europeans have small teeth with comparatively large anterior dimensions. Asian and sub-Saharan African samples share features of average crown size but large cheek teeth. Indian and European samples show considerable overlap on both canonical axes, with average size overall but comparatively large anterior teeth. The few Amerindian samples are too variable to characterize. Based on comparisons of archaeological and living samples, tooth size reductions are documented here for Europe, India, and the Near East compared to tooth sizes of Neolithic and Mesolithic samples. The temporal changes parallel those documented elsewhere for the permanent dentition. The biological and anthropological relevance of these distributions is discussed. PMID- 15895433 TI - Population continuity vs. discontinuity revisited: dental affinities among late Paleolithic through Christian-era Nubians. AB - The present study revisits a subject that has been a source of long-standing bioarchaeological contention, namely, estimation of Nubian population origins and affinities. Using the Arizona State University dental anthropology system, frequencies of 36 crown, root, and intraoral osseous discrete traits in 12 late Pleistocene through early historic Nubian samples were recorded and analyzed. Specifically, intersample phenetic affinities, and an indication of which traits are most important in driving this variation, were determined through the application of correspondence analysis and the mean measure of divergence distance statistic. The results support previous work by the author and others indicating that population discontinuity, in the form of replacement or significant gene flow into an existing gene pool, occurred sometime after the Pleistocene. This analysis now suggests that the break occurred before the Final Neolithic. Samples from the latter through Christian periods exhibit relative homogeneity, which implies overall post-Pleistocene diachronic and regional population continuity. Yet there are several perceptible trends among these latter samples that: 1) are consistent with documented Nubian population history, 2) enable the testing of several existing peopling hypotheses, and 3) allow the formulation of new hypotheses, including a suggestion of two post-Pleistocene subgroups predicated on an age-based sample dichotomy. PMID- 15895434 TI - Brief communication: mtDNA variation in North Cameroon: lack of Asian lineages and implications for back migration from Asia to sub-Saharan Africa. AB - The hypervariable region-1 and four nucleotide positions (10400, 10873, 12308, and 12705) of the coding region of mitochondrial DNA (mtDNA) were analyzed in 441 individuals belonging to eight populations (Daba, Fali, Fulbe, Mandara, Uldeme, Podokwo, Tali, and Tupuri) from North Cameroon and four populations (Bakaka, Bassa, Bamileke, and Ewondo) from South Cameroon. All mtDNAs were assigned to five haplogroups: three sub-Saharan (L1, L2, and L3), one northern African (U6), and one European (U5). Our results contrast with the observed high frequencies of a Y-chromosome haplogroup of probable Asian origin (R1*-M173) in North Cameroon. As a first step toward a better understanding of the evident discrepancy between mtDNA and Y-chromosome data, we propose two contrasting scenarios. The first one, here termed "migration and asymmetric admixture," implies a back migration from Asia to North Cameroon of a population group carrying the haplotype R1*-M173 at high frequency, and an admixture process restricted to migrant males. The second scenario, on the other hand, temed "divergent drift," implies that modern populations of North Cameroon originated from a small population group which migrated from Asia to Africa and in which, through genetic drift, Y-chromosome haplotype R1*-M173 became predominant, whereas the Asian mtDNA haplogroups were lost. PMID- 15895435 TI - Peptide hairpins with strand segments containing alpha- and beta-amino acid residues: cross-strand aromatic interactions of facing Phe residues. AB - The incporation of beta-amino acid residues into the strand segments of designed beta-hairpin leads to the formation of polar sheets, since in the case of beta peptide strands, all adjacent carbonyl groups point in one direction and the amide groups orient in the opposite direction. The conformational analysis of two designed peptide hairpins composed of alpha/beta-hybrid segments are described: Boc-Leu-betaPhe-Val-(D)-Pro-Gly-Leu-betaPhe-Val-OMe (1) and Boc-betaLeu-Phe betaVal-D-Pro-Gly-betaLeu-Phe-betaVal-OMe (2). A 500-MHz 1H-NMR (nuclear magnetic resonance) analysis in methanol supports a significant population of hairpin conformations in both peptides. Diagnostic nuclear Overhauser effects (NOEs) are observed in both cases. X-ray diffraction studies on single crystals of peptide 1 reveal a beta-hairpin conformation in both the molecules, which constitute the crystallographic asymmetric unit. Three cross-strand hydrogen bonds and a nucleating type II' beta-turn at the D-Pro-Gly segment are observed in the two independent molecules. In peptide 1, the betaPhe residues at positions 2 and 7 occur at the nonhydrogen-bonding position, with the benzyl side chains pointing on opposite faces of the beta-sheet. The observed aromatic centroid-to-centroid distances are 8.92 A (molecule A) and 8.94 A (molecule B). In peptide 2, the aromatic rings must occupy facing positions in antiparallel strands, in the NMR derived structure. Peptide 1 yields a normal "hairpin-like" CD spectrum in methanol with a minimum at 224 nm. The CD spectrum of peptide 2 reveals a negative band at 234 nm and a positive band at 221 nm, suggestive of an exciton split doublet. Modeling of the facing Phe side chains at the hydrogen-bonding position of a canonical beta-hairpin suggests that interring separation is approximately 4.78 A for the gauche+ gauche- (g+ g-) rotamer. A previously reported peptide beta-hairpin composed of only alpha-amino acids, Boc-Leu-Phe-Val D-Pro-Gly-Leu-Phe-Val-OMe also exhibited an anomalous far-UV (ultraviolet) CD (circular dichroism) spectrum, which was interpreted in terms of interactions between facing aromatic chromophores, Phe 2 and Phe 7 (C. Zhao, P. L. Polavarapu, C. Das, and P. Balaram, Journal of the American Chemical Society, 2000, Vol 122, pp. 8228-8231). PMID- 15895436 TI - Dental caries among the pre-Hispanic population from Gran Canaria. AB - The island of Gran Canaria was inhabited in pre-Hispanic times by people of North African origin who arrived on the island towards the second half of the first millennium BC. In previous studies, we reported that there are some differences between the coastal inhabitants from Gran Canaria, mainly buried in tumuli, and those from the central mountains, mainly buried in caves. For example, the prevalence of auricular exostoses is higher among the population interred in coastal tumuli when compared with the inland population. This finding may be explained by the practice of marine activities, as supported by chroniclers' reports, by archaeological data, and by bone trace-element analysis, which point to a more intense consumption of marine products by the inhabitants of the coastal regions. Dental caries epidemiology is one of the most important ways in which the diet of past populations can be reconstructed. The purpose of this study is to assess the prevalence and intensity of caries among the pre-Hispanic population of Gran Canaria, in order to increase our knowledge about the dietary habits of these people, and to search for differences among the population buried in caves and that buried in tumuli. We studied a total of 791 individuals. Sex was estimated in 561 cases, and age at death, following Brothwell's criteria, in 549. We found that 66.95% of individuals buried in caves and 58.91% of those buried in tumuli showed at least one carious lesion. The proportion of carious teeth was significantly higher among the population buried in caves (median = 15.71%, interquartile range (IR) = 0-33.33%) than among the population buried in tumuli (median = 6.25%, IR = 0-20%, P = 0.001). Type of burial is the main factor associated with the proportion of carious teeth.These data suggest that the population buried in caves had a different dietary pattern from that of those buried in tumuli. PMID- 15895437 TI - Testing conditional independence in diagnostic palaeoepidemiology. AB - Leprosy was a well-known and dreaded disease in the Middle Ages. A substantial fraction of the adult population carried leprosy-related lesions. Previous research analyzed the occurrence and implications of seven such lesions in samples of medieval skeletons. These analyses were carried out under the assumption of conditional independence among lesion scores. The present paper examines this assumption by developing a test based on the odds ratios and applying the test to three rural medieval samples from Europe: Tirup from the 12th-14th century AD in Jutland, Denmark; Refshale from the 12th-14th century AD on the island of Lolland, Denmark; and Lauchheim from AD 460-680 in southern Germany. Signs of nonzero prevalence of leprosy at death were found in all three samples: Tirup, 25.5% (95% CI, 17.2-34.6%); Refshale, 39.1% (95% CI, 25.5-54.7%); and Lauchheim, 16.2% (95% CI, 10.0-22.9%). It is shown that when leprosy is the prime factor creating variation in the lesion scores in and between samples, the assumption of conditional independence cannot be rejected. PMID- 15895438 TI - Quantification of T-cell-mediated apoptosis in heterogeneous leukemia populations using four-color multiparameter flow cytometry. AB - BACKGROUND: The unique capacity of dendritic cells to present antigens to naive T cells is being increasingly utilized in cancer therapy. The efficacy of cell based immunotherapy can be analyzed by determination of cytotoxic activity of T cells toward tumor cells in vitro. This study supplies a flow cytometric method to analyze T-cell-mediated cytotoxic activity toward heterogeneous leukemic cell populations at a single-cell level. METHODS: The fluorescent probe SYTO16 and the dead-cell dye 7-aminoactinomycine-D (7-AAD) were used to identify early and late stages of apoptosis in combination with leukemia cell-identifying markers. Determination of viable, apoptotic, and necrotic cells was performed by inclusion of fluorescent beads. RESULTS: In nine acute myeloid leukemia samples and three leukemic cell lines the use of SYTO16 next to the dead-cell marker 7-AAD significantly increased (P = 0.001) the sensitivity of the cytotoxicity assay as compared with single use of 7-AAD. Analysis of several effector-to-target ratios revealed the ability to determine dose-response effects. Enumeration of absolute numbers resulted in coefficients of variation of 4.1% and 8.4% for cell lines and leukemic samples, respectively. CONCLUSION: The presented flow cytometric cytotoxicity assay enables the study of T-cell-mediated apoptosis in a heterogenous leukemia population. PMID- 15895439 TI - Growth and nutritional status of male adolescent laborers in Ankara, Turkey. AB - Undernutrition, pathogenic agents, and poor living conditions are of primary importance in the evaluation of adverse environmental conditions' effects on human growth; but child labor (an equally significant factor, especially in underdeveloped countries) is generally overlooked or ignored. The aim of this study is to focus on this subject and clarify the effects of labor on the physical growth and nutritional status of child and adolescent laborers. In this study, the height and weight of 532 male adolescent laborers aged 13.5-18.5 years and their non-laboring peers (n = 451) (the control group) were measured by standard anthropometric techniques and equipment. The individuals of both groups come from lower socioeconomic strata and share similar living conditions. Data were transformed to z-scores, using the US Center for Disease Control and Prevention's 2000 growth charts. The analyses show that the z-scores for height for-age, weight-for-age, and body mass index (BMI)-for-age were negative in both groups. The z-scores of laborers' height-for-age and weight-for-age values lie below the controls', but there is no significant difference between the two groups' BMI-for-age scores. In the laboring group, the percentages of stunting ( 2 SD of height-for-age), underweight (-2 SD of weight-for-age), and wasting (-2 SD of BMI-for-age) were 14.3, 2.6, and 0.2, respectively. These values suggest that malnutrition is not a common problem among adolescent laborers living in Ankara; but laboring is an important cause of faltering in growth, particularly in linear growth, in less or underdeveloped economic environments. PMID- 15895440 TI - Gastric GI stromal tumors (GISTs): the role of surgery in the era of targeted therapy. AB - Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasm arising in the stomach. These tumors were previously classified as smooth muscle tumors, but in recent years it has become clear that they are clinically, pathologically, and molecularly distinct from other tumors and are much more common than previously appreciated. Historically, patients with primary localized or advanced GIST have been managed surgically, as there was no proven role of other treatment modalities such as radiation or chemotherapy. However, the field of GIST was revolutionized with the 1998 discovery that the vast majority of these tumors have oncogenic gain-of-function mutations of the KIT receptor tyrosine kinase. Follow-up studies have confirmed that KIT is both a useful diagnostic marker and an excellent therapeutic target. Imatinib, an inhibitor of KIT kinase activity, is now the standard front-line therapy for patients with advanced GIST. In this review, we discuss pathological and molecular features of gastric GISTs and review the historic and current roles of surgery in the treatment of patients with primary or metastatic GIST. The importance of a multi disciplinary approach using both surgery and imatinib therapy is emphasized. PMID- 15895442 TI - Minimally invasive treatment for gastric cancer: approaches and selection process. AB - Minimally invasive treatment of gastric cancer has emerged as a result of the technical advances, better understanding of gastric physiology, and more knowledge of the biologic behavior of gastric cancer. This treatment results in improved quality of life embodied by smaller incisions, reduced length of hospital stay, and a faster return to productive life. However, minimally invasive treatment for gastric cancer must take into consideration the potential effects of these techniques on tumor dissemination at the time of the treatment procedure, as well as the rates of recurrence and overall survival. Several technical treatment approaches to gastric cancer have now become possible, utilizing endoscopy, laparoscopy, or an open method. Endoscopic mucosal resection (EMR), limited resection, and laparoscopic surgical resection are the currently practiced modalities as the minimally invasive treatment. Lymph node dissection with the minimally invasive techniques is a barrier to its wide application. Although it is not commonly performed in Western countries, the use of minimally invasive treatment for gastric cancer is growing, especially in Korea and Japan. Minimally invasive treatment for early gastric cancer (EGC) has already been shown to be safe and effective in many retrospective series though no prospective randomized studies comparing it to open resection have been performed. Therefore, routine implementation of these procedures must await confirmatory outcomes generated by well-done randomized prospective clinical trials. PMID- 15895444 TI - Perioperative adjunctive treatment in the management of operable gastric cancer. AB - Outcome in the management of clinically resectable gastric carcinoma has been disappointing, at least in Western populations, despite increasingly radical surgery and extensive experience with adjunctive perioperative treatment with innumerable single and combined modality regimens. The United States Intergroup Study, a prospective, randomized, controlled trial of adjuvant chemoradiation, demonstrated significant improvement in disease-free and overall survival. Consequently, this regimen of postoperative fluoruracil plus leucovorin and locoregional radiation has been incorporated into current clinical practice. In hopes of further improving cure rates, many other regimens are under investigation, including the efficacy of neoadjuvant therapy alone, combined neoadjuvant and adjuvant therapy, and adjuvant therapy alone. In these clinical trials, therapeutic agents are prescribed alone or in multimodal regimens and include systemic chemotherapy, intraperitoneal (IP) chemotherapy with or without hyperthermia, intraoperative radiotherapy (IORT), and postoperative external beam irradiation. Several molecular markers have been identified, which seem to predict that a given tumor may be effective or resistant to a drug, raising the possibility of customized chemotherapy regimens. Preclinical studies suggest potential efficacy of angiogenesis inhibitors, monoclonal antibodies, and antisense agents. PMID- 15895446 TI - Eliminating and suppressing local-regional disease in gastric cancer. PMID- 15895448 TI - Status of extended lymph node dissection: locoregional control is the only way to survive gastric cancer. AB - There are many factors that are of influence on gastric cancer treatment. The only way to survive is complete locoregional control. More extended dissections should lead to better outcome, but increased morbidity and mortality probably offset its long-term effect in survival in randomised studies. In this article the factors of influence on outcome of gastric cancer treatment such as the extent of lymph node dissection, splenectomy, pancreatectomy, age, volume and additional treatments are discussed. A literature review of these factors in relation to the latest results of the Dutch Gastric Cancer Trials are presented. If morbidity and mortality can be reduced there might be an advantage of extended lymph node dissection. Splenectomy and pancreatectomy should be performed only in case of direct in growth from the tumour into these organs. Centralisation of gastric cancer treatment should be achieved in order to improve results and to facilitate research. By refining selection criteria in the treatment of gastric cancer further improvements are to be expected. PMID- 15895449 TI - Role of post-operative chemoradiation in resected gastric cancer. AB - The curative management of gastric adenocarcinoma depends upon complete resection of the primary tumor. In patients with lymph node metastases in the resected specimen, the relapse and death rates from recurrent cancer are at least 70%-80%. There is continued debate over whether more extensive lymph node dissection (D2) improves survival when compared to less extensive operations. Until recently, attempts at preventing recurrence have employed adjuvant chemotherapy and have been ineffective. A large U.S. Intergroup study (INT-0116) demonstrated that combined chemoradiation following complete gastric resection improves median time to relapse (30 vs. 19 months, P < 0.0001) and overall survival (35 vs. 28 months, P = 0.01). The improvements in disease-free and overall survival resulting from post-operative chemoradiation have defined a new standard of care. An update of the results of INT-0116 analysis performed in 2004 with 7 years median follow-up, not only confirms the benefits from post-operative chemoradiation but also shows that chemoradiation does not produce significant long-term toxicity. The recent publication of the first large adequately powered III neoadjuvant chemotherapy trial suggested this technique might down-stage tumors and increase resectability. Future advances in the therapy of resectable gastric cancer may come from studies of pre-operative neoadjuvant chemoradiation and the application of targeted therapies such as growth receptor antagonists and anti-angiogenesis agents. PMID- 15895450 TI - Recent advances in sentinel node navigation for gastric cancer: a paradigm shift of surgical management. AB - The sentinel node (SN) is defined as the first draining node from the primary lesion and it has proven to be a good indicator of the metastatic status of regional lymph nodes in solid tumors. In the past 5 years, the validity of SN concept for gastric cancer has been demonstrated by a number of single institutional studies. Using a dual tracer method as the optimal procedure, the radio-guided method allows us to confirm the complete harvest of SNs by gamma probing, while the dye procedure enables us to perform real time observation of the lymphatic vessels. From previous reports, clinically staged T1N0 gastric cancer seems to be a good entity on which to try to change the therapeutic approach based on SN biopsy. At present, two large scale prospective multicenter trials are on-going in Japan. To overcome some remaining issues, such as limited sensitivity of intra-operative diagnosis of metastasis, and technical difficulty in laparoscopic SN detection, further technical and instrumental developments will be required. During this transitional phase, focused lymph node dissection targeted to sentinel lymphatic basins and modified resection of the stomach is an acceptable approach. Although there are several remaining issues, SN navigation should provide a paradigm shift for the surgical management of early gastric cancer in near future. PMID- 15895452 TI - Biologic and clinical variations of adenocarcinoma at the esophago-gastric junction: relevance of a topographic-anatomic subclassification. AB - A topographic-anatomic subclassification of adenocarcinomas of the esophago gastric junction (AEG) in distal esophageal adenocarcinoma (AEG Type I), true carcinoma of the cardia (AEG Type II), and subcardial gastric cancer (AEG Type III) was introduced in 1987 and is now increasingly accepted and used worldwide. Our experience with now more than 1,300 resected AEG tumors indicates that the subtypes differ markedly in terms of surgical epidemiology, histogenesis and histomorphologic tumor characteristics. While underlying specialized intestinal metaplasia can be found in basically all patients with AEG Type I tumors, this is uncommon in Type II tumors and virtually absent in Type III tumors. Stage distribution and overall long-term survival after surgical resection also shows marked differences between the AEG subtypes. Surgical treatment strategies based on tumor type allow a differentiated approach and result in survival rates superior to those reported with other approaches. The subclassification of AEG tumors thus provides a useful tool for the selection of the surgical procedure and allows a better comparison of treatment results. PMID- 15895454 TI - New strategies for the prevention of gastric cancer: Helicobacter pylori and genetic susceptibility. AB - Recently acquired knowledge points to the potential of markedly improved strategies for the prevention of gastric cancer. The International Agency for Cancer Research has reached the conclusion that infection with Helicobacter pylori (H. pylori) is carcinogenic to humans (Group I). The bacterium displays marked genetic heterogeneity. Virulence related genes, especially cag A and vac A s1 m1 are associated with an increased cancer risk. Genetic susceptibility, especially polymorphisms of the cytokine genes may increase cancer risk. Highly susceptible individuals infected with high virulence bacterial genotypes have a markedly increased gastric cancer risk. They should be targeted for endoscopic monitoring to detect advanced precancerous lesions. The state of the art is briefly reviewed and the needed research identified. PMID- 15895456 TI - Walter Lawrence, Jr.: a tribute to a surgical oncologist. "Been there, done that". PMID- 15895459 TI - Gastric cancer: new genetic developments. AB - Gastric cancer's (GC) incidence shows large geographic differences worldwide with the lowest rates occurring in most Western industrialized countries including the United States and the United Kingdom; in contrast, relatively high rates of GC occur in Japan, Korea, China, and South America, particularly Chile. The Lauren classification system classifies GC under two major histopathological variants: 1) an intestinal type and 2) a diffuse type. The intestinal type is more common in the general population, more likely to be sporadic and related to environmental factors such as diet, particularly salted fish and meat as well as smoked foods, cigarette smoking, and alcohol use. It exhibits components of glandular, solid, or intestinal architecture, as well as tubular structures. On the other hand, the diffuse type is more likely to have a primary genetic etiology, a subset of which, known as hereditary diffuse gastric cancer (HDGC), is due to the E-cadherin (CDH1) germline mutation. The diffuse type pathology is characterized by poorly cohesive clusters of cells which infiltrate the gastric wall, leading to its widespread thickening and rigidity of the gastric wall, known as linitis plastica. Helicobacter pylori infection is associated with risk for both the intestinal and diffuse varieties of gastric cancer. Germline truncating mutations of the CDH1 gene, which codes for the E-cadherin protein, were initially identified in three Maori families from New Zealand that were predisposed to diffuse GC. Since then, similar mutations have been described in more than 40 additional HDGC families of diverse ethnic backgrounds. It is noteworthy that two-thirds of HDGC families reported to date have proved negative for the CDH1 germline mutation. A number of candidate genes have been identified through analysis of the molecular biology of E-cadherin. Patients with evidence of the CDH1 germline mutation in the context of a family history of HDGC must be considered as candidates for prophylactic gastrectomy, given the extreme difficulty in its early diagnosis and its exceedingly poor prognosis when there is regional or distant spread. Specifically, the E-cadherin cytoplasmic tail interacts with catenins, assembling the cell-adhesion complex involved with E cadherin mediated cell:cell adhesion. Beta-catenin and gamma-catenin compete for the same binding site on the E-cadherin cytoplasmic tail, directly linking the adhesion complex to the cytoskeleton through alpha-catenin. Beta-catenin gene (CTNNB1) mutations have been described predominantly in intestinal-type gastric cancers and CTNNB1 gene amplification and overexpression have recently been described in a mixed-type gastric cancer. This paper reviews the genetics of both intestinal and diffuse types of gastric carcinoma, their differential diagnosis, molecular genetics, pathology, and, when known, their mode of genetic transmission within families. PMID- 15895460 TI - Determination of variants in the 3'-region of the tyrosinase gene requires locus specific amplification. AB - Mutations in the Tyrosinase gene (TYR, 11q14-q21) cause oculocutaneous albinism type 1 (OCA1). The 3'-region of the TYR shows 98.55% sequence identity with a pseudogene, known as Tyrosinase-Like Gene (TYRL, 11p11.2-cen). A large number of publicly available nucleotide variants of TYR in this region are same as the bases present in the identical locations in the pseudogene. PCR amplification of these regions using primers with sequences common to both loci may result in coamplification of TYR and TYRL, and may lead to misinterpretation of the results. We have resolved this potential problem using locus-specific amplification conditions that could be used to identify unequivocally mutations and SNPs in exon 4 and exon 5 of TYR and proximal flanking sequences. PMID- 15895461 TI - TNF polymorphisms in Alzheimer disease and functional implications on CSF beta amyloid levels. AB - Alzheimer disease (AD), vascular dementia, and stroke are all associated with inflammation though their respective initiating factors differ. Recently a polymorphism in the proinflammatory cytokine tumor necrosis factor (TNF), in association with apolipoprotein E (APOE), was reported to increase AD risk. Two SNPs, rs1799724 (-850C>T; NT_007592.14:g.22400733C>T) and rs1800629 (-308G>A; [NT_007592.14:g.22401282G>A]), and the APOE polymorphism were genotyped in 506 patients with sporadic AD and in 277 cognitively healthy controls. In a subset of 90 individuals we also investigated whether these SNPs exerted any functional effects on cerebrospinal fluid (CSF) beta-amyloid (Abeta) levels. The frequency of the rs1799724 genotypes and the rs1799724-T allele were significantly different in AD individuals (P=0.009; odds ratio [OR], 1.63; 95% confidence interval [CI], 1.13-2.34), while the rs1800629 SNP was not associated with AD. Significant interaction was observed between the rs1799724-T and APOE epsilon4 alleles in that the rs1799724-T allele significantly modified risk associated with possession of the epsilon4 allele only (epsilon4 in absence of rs1799724-T: OR, 2.92; 95% CI, 2.00-4.27; epsilon4 in presence of rs1799724-T: OR, 6.65; 95% CI, 3.26-13.55; P=0.03). Haplotyping analysis revealed a significant overrepresentation of an rs1799724-T/rs1800629-G haplotype in AD (P=0.012; OR, 1.60; 95% CI, 1.11-2.29), although to a lesser degree than rs1799724-T alone. Further, the rs1799724-T allele was found to be associated with lower levels of CSF Abeta42 (P=0.023), thus corroborating the genetic findings. Inheritance of the rs1799724-T allele appears to synergistically increase the risk of AD in APOEepsilon4 carriers and is associated with altered CSF Abeta42 levels. Further investigations are warranted to assess the significance of these novel findings. PMID- 15895462 TI - The phenotypic spectrum of COL2A1 mutations. AB - Heterozygous mutations of COL2A1 create several clinical entities collectively termed type II collagenopathies. These disorders not only impair skeletal growth but also cause ocular and otolaryngological abnormalities. The classical phenotypes include the spondyloepiphyseal dysplasia (SED) spectrum with variable severity, Stickler dysplasia type I (STD-I), and Kniest dysplasia (KND). Most COL2A1 mutations occur in the triple helical region of alpha 1(II) chains: the SED spectrum is mostly attributed to missense mutations that substitute bulky amino acids for glycine residues, STD-I to haploinsufficiency of truncation mutations, and KND to exon skipping due to splice-site mutations. To further elucidate the genotype-phenotype relationship of type II collagenopathies, we examined COL2A1 mutations in 56 families that were suspected of having type II collagenopathies, and found 38 mutations in 41 families. Phenotypes for all 22 missense mutations and one in-frame deletion in the triple helical region fell along the SED spectrum. Glycine to serine substitutions resulted in alternating zones that produce severer and milder skeletal phenotypes. Glycine to nonserine residue substitutions exclusively created more severe phenotypes. The gradient of the SED spectrum did not necessarily correlate with the occurrence of extraskeletal manifestations. All nine truncation or splice-site mutations in the triple helical or N-propeptide region caused STD-I or KND, and extraskeletal changes were inevitable in both phenotypes. All six C-propeptide mutations produced a range of atypical skeletal phenotypes and created ocular, but not otolaryngological, changes. PMID- 15895463 TI - Quantitative validation of different protein precipitation methods in proteome analysis of blood platelets. AB - For the preparation of proteins for proteome analysis, precipitation is frequently used to concentrate proteins and to remove interfering compounds. Various methods for protein precipitation are applied, which rely on different chemical principles. This study compares the changes in the protein composition of human blood platelet extracts after precipitation with ethanol (EtOH) or trichloroacetic acid (TCA). Both methods yielded the same amount of proteins from the platelet preparations. However, the EtOH-precipitated samples had to be dialyzed because of the considerable salt content. To characterize single platelet proteins, samples were analyzed by two-dimensional fluorescence differential gel electrophoresis. More than 90% of all the spots were equally present in the EtOH- and TCA-precipitated samples. However, both precipitation methods showed a smaller correlation with nonprecipitated samples (EtOH 74.9%, TCA 79.2%). Several proteins were either reduced or relatively enriched in the precipitated samples. The proteins varied randomly in molecular weight and isoelectric point. This study shows that protein precipitation leads to specific changes in the protein composition of proteomics samples. This depends more on the specific structure of the protein than on the precipitating agent used in the experiment. PMID- 15895464 TI - A sensitive and rapid immunoassay for quantification of CA125 in human sera by capillary electrophoresis with enhanced chemiluminescence detection. AB - In this paper we have presented a sensitive and rapid immunoassay (IA) method by capillary electrophoresis with an enhanced chemiluminescence detection system (CE CL) based on the catalytic effects of horseradish peroxidase (HRP) on the luminol hydrogen peroxide reaction. The conditions for the CL reaction and electrophoresis were systematically investigated using HRP as a model sample. The linear range from 2.5 x 10(-11) to 1.0 x 10(-9) mol/L (R = 0.999), and the detection limit of 1.0 x 10(-12) mol/L (signal-to-noise ratio = 3) for HRP were achieved using para-iodophenol as CL enhancer. The relative standard deviations of the migration time and peak area for 5.0 x 10(-10) mol/L HRP (n = 7) were 0.26 and 4.8%, respectively, using a CE system with a home-built CL detector. Under the optimal condition, the HRP-labeled CA125 antibody (Ab) and the Ab-antigen complex were well separated within 4 min by CE using a high-pH buffer (pH 10.20). The assay was successfully used for quantification of CA125 in human sera from health controls and patients associated with ovarian cancer, and the recoveries of the standard addition experiments were 93-109%. Our primary results demonstrated that IA based on CE-CL detection is a powerful tool for clinical diagnosis combined with these commercial IA kits. PMID- 15895465 TI - An ultrasensitive method for the determination of thiouracil and phenylthiouracil using capillary zone electrophoresis and laser-induced fluorescence detection. AB - This paper reports the development of a method based on capillary electrophoresis with laser-induced fluorescence detection for the simultaneous determination of thiouracil (TU) and phenylthiouracil (PhTU) with high sensitivity (nanomolar range, i.e., attomoles detected). After derivatization with 5 iodoacetamidofluorescein, the analytes were separated by capillary zone electrophoresis using 20 mM phosphate buffer (pH 10.0) and quantified by fluorescence detection. The linearity range, precision, recovery, and detection limits were determined, and the method was shown to be applicable for the determination of TU and PhTU in spiked feed samples and urine. PMID- 15895466 TI - Molecular control of mitochondrial function in preimplantation mouse embryos. AB - Mitochondria play a key role in a number of physiological events during all stages of life, including the very first stages following fertilization. It is, therefore, important to understand the mechanisms controlling mitochondrial activity during early embryogenesis to determine their role in development outcome. The objective of this study was to investigate the molecular control of mitochondrial transcription and mitochondrial DNA (mtDNA) replication in mouse preimplantation embryos. We estimated the mtDNA copy number and characterized the expression patterns of two mitochondrial genes and several nuclear genes that encode mitochondrial transcription and replication factors throughout preimplantation development. Mitochondrial gene transcripts were present in larger quantities in morula and blastocyst stage embryos relative to other stages. A significant increase in the amount of mRNA for nuclear genes encoding mtDNA transcription factors was observed in eight-cell stage embryos. Although a similar increase in the mRNA levels of nuclear genes encoding mtDNA replication factors was observed in morula and blastocyst stage embryos, the number of mtDNA molecules remained stable during preimplantation stages, suggesting that nuclear encoded mitochondrial transcription factors are involved in the regulation of mtDNA transcription during early development. Although transcripts of replication factors are abundant at the morula and blastocyst stage, mtDNA replication did not occur until the blastocyst stage, suggesting that the inhibition of mtDNA replication is controlled at the post-transcriptional level during early embryogenesis. PMID- 15895467 TI - Decreasing analysis time in capillary electrophoresis: validation and comparison of quantitative performances in several approaches. AB - Capillary electrophoresis (CE) can be used for the rapid determination of pharmaceuticals, particularly in routine quality control analysis. This paper focuses on several approaches aimed at decreasing the analysis time with commercially available instrumentation by (i) application of a high electric field through a reduced capillary, (ii) use of a dynamically coated capillary to increase the electroosmotic flow, (iii) short-end injection (SEI) technique, and (iv) application of multiple sample injections. Moreover, SEIs were combined with the three other approaches. A pharmaceutical formulation containing lidocaine as an active component was selected, and the methods were validated according to the ICH guidelines. The seven approaches investigated fulfilled different statistical requirements and demonstrated their linearity and trueness, with good recoveries and confidence limits always inferior to 1.5%. Furthermore, relative standard deviation (RSD) values for repeatability and intermediate precision were inferior to 1.1 and 1.8%, respectively. These results confirmed that each approach is of utmost interest to increase the analyte throughput in CE. PMID- 15895468 TI - Search for peptide sequences involved in human antisperm antibody-mediated male immunoinfertility by using phage display technology. AB - The aim of the present study was to delineate peptide sequences against which antisperm antibodies (ASA) are raised in immunoinfertile men. Using the phage display technology, seven unique and novel dodecamer amino acid sequences were identified that reacted with the sera of immunoinfertile men. The peptides were synthesized based upon these amino acid sequences and examined for their immunoreactivity with sera from ASA-positive immunofertile men (n = 15) and ASA negative fertile men (n = 18) for IgM, IgG, and IgA class of antibody in the enzyme-linked immunosorbent assay (ELISA). All the seven synthetic peptides showed a significantly (P < 0.001) higher mean absorbance values for IgG and/or IgA class of antibody with the immunoinfertile sera compared to fertile control sera. Three of the seven peptides demonstrated a stronger reaction (>2 SD units) with 27%-40% of immunoinfertile sera compared to fertile controls. These peptide sequences may find applications in the specific diagnosis and treatment of immunoinfertility and in contraceptive vaccine development. The phage display technique provides an exciting and novel technology to delineate sperm epitopes involved in immunoinfertility. PMID- 15895469 TI - Expression of imprinted genes is aberrant in deceased newborn cloned calves and relatively normal in surviving adult clones. AB - Cattle are the species used most frequently for the development of assisted reproductive technologies, such as nuclear transfer. Cattle cloning can be performed by a large number of laboratories around the world, and the efficiency of nuclear transfer in cattle is the highest among all species in which successful cloning has been achieved. However, an understanding of the expression of imprinted genes in this important species is lacking. In the present study, real time reverse transcription polymerase chain reaction (RT-PCR) was utilized to quantify the expression of the bovine Igf2, Igf2r, and H19 genes in eight major organs (brain, bladder, heart, kidney, liver, lung, spleen, and thymus) of somatic cell cloned calves that died shortly after birth, in three tissues (skin, muscle, and liver) of healthy clones that survived to adulthood, and in corresponding tissues of control animals from natural reproduction. We found that, deceased bovine cloned calves exhibited abnormal expression of all three genes studied in various organs. Large variations in the expression levels of imprinted genes were also seen among these clones, which were produced from the same genetic donor. In surviving adult clones, however, the expression of these imprinted genes was largely normal, except for the expression of the Igf2 gene in muscle, which was highly variable. Our data showed disruptions of expression of imprinted genes in bovine clones, which is possibly due to incomplete reprogramming of donor cell nuclei during nuclear transfer, and these abnormalities may be associated with the high neonatal mortality in cloned animals; clones that survived to adulthood, however, are not only physically healthy but also relatively normal at the molecular level of those three imprinted genes. PMID- 15895470 TI - Hair analysis by GC/MS/MS to verify abuse of drugs. AB - Because of its peculiar characteristics, hair analysis provides a way of obtaining information that cannot be acquired by other commonly used forensic medical analyses, such as blood or urine analysis. In the keratin matrix many xenobiotics are incorporated permanently, in contrast to the situation with blood or urine where they are generally only detectable for a few hours or days. Therefore hair analysis should be the method of choice in the clinical and forensic toxicology field when the assessment of repeated or chronic exposure to a drug is required, e.g. in the case of criminal responsibility, revocation/restoration of a driving licence or in workplace testing. Some factors that can affect the concentrations of drugs in hair, such as passive contamination, age, ethnicity and cosmetic treatment, must be considered. Analytical methodology is also very important: GC/MS/MS has proved to be a highly sensitive and specific technique for the detection of very low concentrations of such drugs in hair. In this study five cases of the application of hair analyses using this technique for the determination of abused drugs (opiates, cocaine, amphetamine, anabolic steroids) are described. PMID- 15895471 TI - Glycopeptide and glycoprotein synthesis involving unprotected carbohydrate building blocks. AB - This review summarizes the chemical and chemoenzymatic synthesis of glycopeptides and glycoproteins using unprotected carbohydrates as key intermediates. The synthetic methods covered herein include the convergent synthesis of glycopeptides by chemoselective ligation of peptides and free glycans, solution- and solid-phase synthesis of glycopeptides by sequential peptide elongation with unprotected glycosyl amino acids or short glycopeptides as building blocks, and the synthesis of glycopeptides by enzymatic and/or chemical elongation of the free glycans. The use of unprotected carbohydrates in these syntheses can circumvent the final-stage carbohydrate deprotection, lead to highly convergent synthetic designs, and more significantly, take advantage of the commercially available free glycans isolated from nature, which could considerably facilitate the synthesis of complex glycopeptides and glycoproteins. PMID- 15895472 TI - Highly branched stimuli responsive poly[(N-isopropyl acrylamide)-co-(1,2 propandiol-3-methacrylate)]s with protein binding functionality. AB - Highly branched poly(NIPAM) have been prepared using the technique of reversible addition-fragmentation chain transfer (RAFT) polymerisation using a chain transfer agent that allows the incorporation of imidazole functionality in the polymer chain-ends. The lower critical solution temperature (LCST) of the polymers can be controlled by the amount of hydrophobe and GMA incorporated during copolymerisation procedures. These thermally responsive "smart" polymers were used to purify a His-tagged BRCA-1 protein fragment by affinity precipitation. [Diagram: see text] PMID- 15895473 TI - Lipopolymers from new 2-substituted-2-oxazolines for artificial cell membrane constructs. AB - We present the synthesis of novel 2-oxazoline monomers with different 2 substituents and their consecutive conversion into lipopolymers by living cationic polymerization. The side functions of these monomers were varied to realize different steric needs and hydrogen bonding interactions of the polymer side chains. 2-(2'-N-pyrrolidonyl-ethyl)-2-oxazoline, 2-(3' methoxymonoethyleneglycol)propyl-2-oxazoline, and 2-(3' methoxytriethyleneglycol)propyl-2-oxazoline were synthesized. All of the monomers could be converted into the corresponding lipopolymers by living cationic polymerization using 2,3-di-O-octadecyl-1-trifluormethansulfonyl-sn-glycerol as the initiator. The characterization of the 2,3-di-O-octadecyl-glycerol-poly(2 oxazoline) lipopolymers by NMR spectroscopy, IR spectroscopy, and gel permeation chromatography revealed that the targeted molar masses and compositions can be controlled by the initial initiator/monomer ([M](0)/[I](0)) ratio for all the synthesized lipopolymers. The polydispersities were found to be narrow (polydispersity indices from 1.06-1.3). The amphiphilic lipopolymers were spread at the air-water interface (Langmuir-Blodgett film balance) and the effect of the polymer side groups and chain lengths upon the Pi-area (A) isotherms of the corresponding lipopolymer monolayers were compared and analyzed. The impact of the polymer side functionalities on a 2D gel formation was examined using an interfacial rheometer operated in an oscillating stress-strain mode. Interestingly enough, none of the newly synthesized lipopolymers showed a rheological transition. This somewhat surprising result not only verified that these 2D gels are not established by hydrogen bonding among hydrophilic polymer moieties, as earlier proposed, but also supported the concept of jammed surface micelles as the more likely origin for the gelation phenomenon. [Diagram: see text] PMID- 15895474 TI - Alkaline phosphatase encapsulated in gellan-chitosan hybrid capsules. AB - Alkaline phosphatase (ALP) was encapsulated in gellan-chitosan polyion complex (PIC) capsules using a convenient procedure. The recovery of ALP was about 50% when the capsules were prepared by dropping a solution of ALP and gellan mixture (ALP/gellan) into a chitosan solution. When p-nitrophenyl phosphate (p-NPP) and 5 bromo-4-chloro-3-indolyl phosphate (BCIP) were incubated with ALP/gellan-chitosan capsules as substrates for ALP, the transparent colorless capsules changed to yellow and blue, respectively. The encapsulation of ALP into the PIC capsules was also confirmed by SDS-PAGE and immunoblot analyses. The ALP and polypeptides of more than 30 kDa remained without release even after incubation at 4 degrees C for 14 d. The biochemical properties of the encapsulated ALP activity were similar to those of the intact enzyme. When the solution containing p-NPP was loaded on a column packed with ALP/gellan-chitosan capsules at 27 degrees C, approximately 75% of p-NPP was hydrolyzed by passing through the column. No significant leakage of ALP was observed during the procedure, indicating that the capsules were resistant to pressure in the chromatographic operation. Furthermore, 70% of the hydrolytic activity of the packed capsules remained after storage at 4 degrees C for one month. These results suggest that the polyion complex capsules could be useful materials for protein fixation without chemical modification. [Diagram: see text] Encapsulation of ALP into PIC capsules and the morphological changes seen in the absence of the ALP substrate and in the presence of p-NPP and BICP. PMID- 15895475 TI - Preparation of alpha-cyclodextrin-terminated polyrotaxane consisting of beta cyclodextrins and pluronic as a building block of a biodegradable network. AB - A beta-CD-based biodegradable polyrotaxane was prepared by capping both terminals of polypseudorotaxane consisting of hydrazide-terminated PEG-block-PPG-block-PEG (Pluronic P-105) and beta-CD-succinates with mono-aldehyde alpha-CDs. By decreasing pH, the fluorescent intensity of TNS was increased with time, indicating cleavage of the terminal hydrazone bonds followed by beta-CD-succinate release. The terminal alpha-CD moieties of the polyrotaxane are useful for self assembled formation with some guest molecules. [Diagram: see text] PMID- 15895476 TI - Vascular endothelial and smooth muscle cell culture on NaOH-treated poly(epsilon caprolactone) films: a preliminary study for vascular graft development. AB - Tissue engineering offers the potential of providing vessels that can be used to replace diseased and damaged native blood vessels. The endothelization of a synthetic vascular graft minimizes the failures associated with blood clotting and platelet activation. The aim of this study was to culture vascular-derived endothelial and smooth muscle cells on both untreated and NaOH-treated poly(epsilon-caprolactone) (PCL) films, a biocompatible and bio-resorbable polymer, and to evaluate the behavior of both cell types as a preliminary study for vascular graft development. PCL films were prepared by hot pressing; characterized by DSC, IR, SEM, and scanning force microscopy; and treated with NaOH to increase the surface hydrophilicity before cell culture. Endothelial and smooth muscle cells, isolated from pig cava vein, were characterized by immunofluorescence and confocal microscopy studies of endothelial nitric oxide synthase and alpha-smooth muscle actin. Good adhesion, growth, viability and morphology of both the endothelial and smooth muscle cells on PCL films were obtained, but a light stimulation of mitochondrial activity was observed during short culture times. NaOH treatment improved the adhesion and enhanced the proliferation in both cell types. This verified the possible use of this modified polymer as a support in the preparation of a synthetic vascular graft. [Diagram: see text] SEM micrograph of smooth muscle cells cultured on NaOH-treated PCL film. (Original magnification: 1000x). PMID- 15895477 TI - Kinetic parameters of urease immobilized on modified acrylonitrile copolymer membranes in the presence and absence of Cu(II) ions. AB - Poly(acrylonitrile-methylmethacrylate-sodium vinylsulfonate) membranes were subjected to seven different chemical modifications and the amount of the newly formed groups was measured for each membrane. Urease was then covalently immobilized onto the modified membranes and the amount of bound protein was determined. The kinetic parameters V(max) and K(m) of the immobilized urease were studied under static and dynamic conditions. Results showed that the rate of the enzyme reaction was higher for the membranes modified with NH(2)OH . H(2)SO(4), NH(2)NH(2) . H(2)SO(4), NaOH + EDA and NaOH + GA + EDA. It was confirmed that the reaction rate, measured under dynamic conditions, was higher than that one determined under static conditions. The influence of Cu(II) ions, as inhibitors, on the enzyme reaction kinetics (V(i) and K(i)) was also investigated. It turned out that the most sensitive membranes towards Cu(II) were those modified with NH(2)NH(2) . H(2)SO(4), NaOH + EDA and H(2)O(2). The results initiated further investigations on the influence of other heavy metal ions (Cd(II), Zn(II), Ni(II) and Pb(II)) over urease bound to a NH(2)OH . H(2)SO(4)-modified membrane. It was found that the inhibition effect of the heavy metal ions over immobilized urease decreases in the order: Cu(II) > Cd(II) > Zn(II) > Ni(II) > Pb(II). [Diagram: see text] PMID- 15895478 TI - Effect of Th2 cytokine antagonist treatments on chemical-induced allergic response in mice. AB - To understand better the cellular and molecular mechanisms of chemical-induced occupational asthma, we examined the effects of the Th2 cytokine antagonists interferon-gamma (IFN-gamma), interleukin (IL)-12 and anti-IL-4 on the balance of the Th1/Th2 response induced by trimellitic anhydride (TMA) and phthalic anhydride (PA). Eight- to ten-week-old BALB/c mice were assigned to be exposed to either TMA or PA plus one of these Th2 cytokine antagonists. Both TMA (25% and 12.5% for sensitization and challenge, respectively) and PA (12.5% and 6.25% for sensitization and challenge, respectively) induced a Th2 response marked by an increasing production of IL-4 and IL-10 in the supernatants of ex vivo spleen cells cultured with concanavalin A and also of serum total IgE. Co-administration of IL-12 and antiIL-4 deviated these PA- or TMA-induced Th2 responses, as judged by an increasing serum total IgG2a production (up to 14-fold), associated with a slight decrease of IL-4 in three out of four experiments and of IL-10 in all four experiments. Co-administration of IFN-gamma, however, had only one weak effect. These findings suggest that the chemical-induced Th2-biased response may be diverted during an induction period by exogenous administration of the Th2 cytokine antagonists, particularly IL-12 and the anti-IL-4 antibody. These results would significantly enhance our understanding of the Th1/Th2 response induced by chemicals. PMID- 15895479 TI - Analysis of opiates, cocaine and metabolites in urine by high-performance liquid chromatography with diode array detection (HPLC-DAD). AB - An analytical method is proposed for the simultaneous determination of morphine, codeine, 6-acetyl-morphine (MAM), cocaine, benzoylecgonine (BEG), cocaethylene, methadone and 2-ethylen-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in urine using high performance liquid chromatography coupled to a diode array detector (HPLC-DAD). The selection of working wavelengths is based on the highest chromatographic response for each component: 233 nm for cocaine, BEG and cocaethylene; 285 nm for morphine, codeine and MAM; and 292 nm for methadone and EDDP. The mobile phase, which is a mixture of acetonitrile and 0.02 M phosphate buffer at pH 6.53, was eluted in gradient mode through an XTerra RP-8 column (250 mm x 4.6 mm i.d., 5 microm particle size). After applying a solid-phase extraction procedure with Bond Elut Certify cartridges, the recoveries obtained were between 60% (EDDP) and 97% (cocaethylene). A good linearity of the method in the 0.1-10 microg mL(-1) range of urinary concentrations was obtained because the coefficient of correlation exceeded 0.99 for each drug. The precision and accuracy were quite good, with values of <7% and within the range +/- 6%, respectively. Finally, the proposed method was applied to 23 urine samples from fatal intoxications related to methadone, heroin and[sol ]or cocaine. PMID- 15895480 TI - Estrogen therapy increases risk of gallstone development. PMID- 15895481 TI - I have bad nighttime leg cramps. What causes this, and what can I do about it? PMID- 15895482 TI - The dosages of some of my vitamins are given in terms of "IU". What is an IU? PMID- 15895483 TI - Efficacy of picroliv in combination with miltefosine, an orally effective antileishmanial drug against experimental visceral leishmaniasis. AB - Visceral leishmaniasis (VL) or kala-azar continues to persist as one of the major public health problems in many tropical countries. However, no effective treatment for radical cure of the disease is yet available. Miltefosine, an alkyl phospholipid compound, is the first orally effective drug, which has shown 98% cure rate of VL patients during phase III clinical trial in India. Since this drug requires long course of treatment and has long half-life, there are fairly good chances of emergence of resistance. Furthermore, this drug has produced severe side-effects in some of the cases. We therefore examined the possibility of minimizing these effects by applying miltefosine in lower doses in combination with picrloviv, an immunomodulator against Leishmania donovani in hamsters (Mesocricetus auratus). The picroliv per se showed no antileishmanaial potential. However, when given with suboptimal dose of miltefosine, it enhanced efficacy of the latter from 45 to 86% on day 7 post treatment and from 32 to 64% on day 28 post treatment. Interestingly, the efficacy of this combination was as good as the curative dose of miltefosine alone. Thus, this combination appears to offer a fruitful strategy for treatment of VL. PMID- 15895484 TI - An unusual Gram stain finding. PMID- 15895485 TI - Emerging into the light: working children's photodiaries. PMID- 15895486 TI - Socioeconomic position (again), causes and confounding. PMID- 15895487 TI - Take allergy drug and sniff flowers safely? PMID- 15895488 TI - Asymptomatic ruptured sinus of Valsalva aneurysm into the right ventricle. PMID- 15895489 TI - Magnetic resonance spectroscopic studies of alcoholism: from heavy drinking to alcohol dependence and back again. PMID- 15895490 TI - Applications of morphometric and diffusion tensor magnetic resonance imaging to the study of brain abnormalities in the alcoholism spectrum. PMID- 15895491 TI - [Schistosomiasis transmission in urban area]. PMID- 15895492 TI - Katherine Austin Lathrop. PMID- 15895493 TI - Healthier arteries, clearer thinking. What you do for your heart pays off for your brain and memory. PMID- 15895494 TI - A protein emerges as a heart threat, blunted by statins. PMID- 15895495 TI - How to choose a health club. As fitness centers sweat to attract you, stay cool and careful. PMID- 15895496 TI - Demystifying metabolic syndrome. How you can counter a hydra-headed menace to your heart. PMID- 15895497 TI - Whole grains: the whole truth. What to look for in food labels to ensure health benefits. PMID- 15895498 TI - Ask the doctors. I have great disparity in my blood pressures, which are generally around 145 over 65 with a pulse of 55. Am I still considered hypertensive? Should I be on medication for high blood pressure? PMID- 15895499 TI - Managing cancer-related fatigue in palliative care. AB - Cancer-related fatigue is the most frequently experienced symptom in palliative care patients suffering from cancer, and has a profound effect on their quality of life. This article explores the current literature relating to the assessment and management of this complex and distressing symptom in patients with advanced cancer. PMID- 15895500 TI - Introducing a hand-held urinal service in secondary care. AB - Chorley & South Ribble Primary Care Trust's multiprofessional continence service was established in 2000 (Pomfret, 2001). An integral part of this service development was the creation of a community-based, hand-held urinal library. There are 28 female hand-held urinals available in the UK and many more for males. Some of the urinals are available on prescription and others have to be purchased by the patient. The urinal library allows patients in the community to use the urinals before they are prescribed or buy them, because it is only by using them that they will know that the product will be suitable. The service is proving to be hugely successful. PMID- 15895501 TI - Space flight and bone formation. AB - Major physiological changes which occur during spaceflight include bone loss, muscle atrophy, cardiovascular and immune response alterations. When trying to determine the reason why bone loss occurs during spaceflight, one must remember that all these other changes in physiology and metabolism may also have impact on the skeletal system. For bone, however, the role of normal weight bearing is a major concern and we have found no adequate substitute for weight bearing which can prevent bone loss. During the study of this problem, we have learned a great deal about bone physiology and increased our knowledge about how normal bone is formed and maintained. Presently, we do not have adequate ground based models which can mimic the tissue loss that occurs in spaceflight but this condition closely resembles the bone loss seen with osteoporosis. Although a normal bone structure will respond to application of mechanical force and weight bearing by forming new bone, a weakened osteoporotic bone may have a tendency to fracture. The study of the skeletal system during weightless conditions will eventually produce preventative measures and form a basis for protecting the crew during long term space flight. The added benefit from these studies will be methods to treat bone loss conditions which occur here on earth. PMID- 15895502 TI - Control of gravitropic orientation. I. Non-vertical orientation by primary roots of maize results from decay of competence for orthogravitropic induction. AB - Plant organs may respond to gravity by vertical (orthogravitropic), oblique (plagiogravitropic) or horizontal (diagravitropic) growth. Primary roots of maize (Zea mays L.) provide a good system for studying such behaviours because they are reportedly capable of displaying all three responses. In current work using maize seedlings of the Silver Queen cultivar, stabilisation of growth at an oblique orientation was commonplace. Hypothetically, plagiogravitropism may be accomplished either by a process we call graded orthogravitropism or by hunting about a sensed non-vertical setpoint. In graded orthotropism primary bending is unidirectional and depends on facilitative stimuli that determine its extent. The hallmark of the setpoint mechanism is restorative curvature of either sign following a displacement; both diagravitropism and orthogravitropism are based on setpoints. Roots settled in a plagiogravitropic orientation were tested with various illumination and displacement protocols designed to distinguish between these two hypotheses. The tests refuted the setpoint hypothesis and supported that of graded orthotropism. No evidence of diagravitropism could be found, thus, earlier claims were likely based on inadequately controlled observations of graded orthotropism. We propose that orthotropism is graded by the sequential action of dual gravity receptors: induction of a vectorial gravitropic response requires gravitational induction of a separate facilitative response, whose decay in the absence of fresh stimuli can brake gravitropism at plagiotropic angles. PMID- 15895503 TI - Control of gravitropic orientation. II. Dual receptor model for gravitropism. AB - Gravitropism of vascular plants has been assumed to require a single gravity receptor mechanism. However, based on the evidence in Part I of this study, we propose that maize roots require two. The first mechanism is without a directional effect and, by itself, cannot give rise to tropism. Its role is quantitative facilitation of the second mechanism, which is directional like the gravitational force itself and provides the impetus for tropic curvature. How closely coupled the two mechanisms may be is, as yet, unclear. The evidence for dual receptors supports a general model for roots. When readiness for gravifacilitation, or gravifacilitation itself, is constitutive, orthogravitropic curvature can go to completion. If not constitutively enabled, gravifacilitation can be weak in the absence of light and water deficit or strong in the presence of light and water deficit. In either case, it can decay and permit roots to assume reproducible non-vertical orientations (plagiogravitropic or plagiotropic orientations) without using non-vertical setpoints. In this way roots are deployed in a large volume of soil. Gravitropic behaviours in shoots are more diverse than in roots, utilising oblique and horizontal as well as vertical setpoints. As a guide to future experiments, we assess how constitutive v. non constitutive modes of gravifacilitation might contribute to behaviours based on each kind of setpoint. PMID- 15895504 TI - Morphological aspects of oligomeric protein structures. AB - Features of multimeric proteins are reviewed to shed light on the formation of protein assemblies from a structural perspective. The features comprise biochemical and geometric properties. They are compiled on new low-redundancy sets of crystal structures of homomeric proteins with different symmetry and subunit multiplicity, as well as on a set of heteromeric proteins. Crystal structures of likely monomers provide a control group. PMID- 15895505 TI - Restaurant foodhandler-associated outbreak of Salmonella Heidelberg gastroenteritis identified by calls to a local telehealth service, Edmonton, Alberta, 2004. PMID- 15895506 TI - Confidential services for teenagers. PMID- 15895507 TI - Statement on personal protective measures to prevent arthropod bites. PMID- 15895508 TI - Update on meningococcal C conjugate vaccines. PMID- 15895509 TI - Statement on travel, influenza, and prevention. PMID- 15895510 TI - Interchangeability of diphtheria, tetanus, acellular pertussis, polio, Haemophilus influenzae type B combination vaccines presently approved for use in Canada for children < 7 years of age. PMID- 15895511 TI - [Cholecysto-cardiac syndrome: a myth or reality?]. PMID- 15895512 TI - Conversion ratio of tryptophan to niacin in Japanese women fed a purified diet conforming to the Japanese Dietary Reference Intakes. AB - In order to establish the human requirements of niacin, it is first important to know how much tryptophan is converted to niacin in the human body. In a general, 60 mg of tryptophan is equivalent to 1 mg of niacin, whereas the conversion ratio of tryptophan to niacin is yet to be confirmed. The aim of this study was to know the conversion ratio of tryptophan to niacin in Japanese females fed a purified diet, which followed the Japanese Dietary Reference Intakes. Ten young Japanese females were housed in the same facility and given the same daily living activity schedule for 7 d. The composition of their purified diet was conformed to the Dietary Reference Intakes in Japan. The diet was niacin free. In order to investigate the conversion ratio, daily urinary outputs were collected. Tryptophan-niacin metabolites in the urine were measured and the conversion ratio of tryptophan to niacin calculated. The conversion ratio was calculated by comparing the dietary intake of tryptophan and the sum of the niacin catabolites such as N1-methylnicotinamide, N1-methyl-2-pyridone-5-carboxamide, and N1-methyl 4-pyridone-3-carboxamide, which were derived only from the dietary intake of tryptophan. The ratio was calculated as 1.5 +/- 0.1 (mean +/-SE for 10 women; in molar basis) on the last day of the experiment. It was calculated that if the excretory percentage of niacin metabolites in the urine were 60%, of the tryptophan ingested, the conversion factor would be a value of 67, meaning that is 67 mg of tryptophan is equal to 1 mg of niacin. PMID- 15895513 TI - Changes in the urinary excretion of the metabolites of the tryptophan-niacin pathway during pregnancy in Japanese women and rats. AB - NAD is biosynthesized from tryptophan. Therefore, experimental studies including tryptophan metabolism studies could provide insight into niacin nutrition in pregnancy. Our aim was to determine the change in niacin metabolism during pregnancy by a systemic investigation of how pregnancy alters the tryptophan niacin metabolism in Japanese women and rats. For the human study, spot urine samples were collected from a total of 434 pregnant Japanese women who were at 5 40 wk of gestation, 50 women at 4-6 wk postpartum, and 10 nonpregnant women as the controls. For the animal study, pregnant rats were fed with a niacin-free diet, and daily urine samples were collected from day 6 of gestation to day 6 postpartum. The intermediates and metabolites of the tryptophan-niacin pathway in the urine samples were measured. The urinary excretions of niacin metabolites in humans and rats increased from mid pregnancy in a time-dependent manner, reached a peak of 2-3-fold during late pregnancy, and declined to control levels after childbirth. PMID- 15895514 TI - The effect of Lactobacillus casei strain Shirota on the cecal fermentation pattern depends on the individual cecal microflora in pigs. AB - Probiotic bacteria improve fermentation in the large intestine through interaction with indigenous bacteria. The microflora in the large intestine differ from one individual to another. The objective of this study was to determine whether the different cecal microflora induce different probiotic effects on cecal fermentation in three pigs (pig A, B and C). Pigs were administered fermented milk prepared with the Lactobacillus casei strain Shirota (LCS) as a probiotic for 2 wk. The average number of LCS was higher than 5.0 (log cfu/g cecal digesta) in all pigs. The hierarchical clustering of the temperature gradient gel electrophoresis (TGGE) profile of cecal bacteria showed that the cecal microflora was slightly altered by the LCS dose in all pigs. However, the molar ratios of cecal butyrate in pigs A and C were significantly increased by LCS. Inversely, the molar ratio of propionate in pigs A and C was significantly decreased by LCS. However, the molar ratio of individual short-chain fatty acid in pig B was not significantly affected by LCS. The hierarchical clustering of the TGGE profiles indicated that the cecal microflora of pig A resembled that of pig C and that of pig B was dissimilar to those of pigs A and C. These results indicated that the effect of LCS on fermentation in the large intestine was possibly dependent on the composition of indigenous microflora. PMID- 15895515 TI - Determination of molybdenum in foods and human milk, and an estimate of average molybdenum intake in the Japanese population. AB - To evaluate the molybdenum (Mo) status in the Japanese population, the Mo content in various foods and human milk was determined using inductively coupled plasma mass spectrometry (ICP-MS) and the average Mo intake was estimated. The difference in Mo content among food groups was marked; Mo levels in several plant foods such as cereals were more than 0.5 g/g while those in most animal foods were less than 0.1 microg/g. In particular, Mo contents in several samples of seeds and pulses were more than mixeo 1 mirog/g. The variation in Mo contents in each type of cereal was also conspicuous. Based on the present quantification of Mo in foods and the recent National Nutrition Survey in Japan, the average Mo intake of the Japanese population was estimated as 225 microg/d/capita. The principal Mo source in the Japanese diet was rice followed by soybean products, and approximately 90% of the Mo intake was derived from plant foods. Seventeen human milk samples were collected from 3 healthy mothers once or twice a month from 96 to 327 d after delivery. The median and range of Mo in human milk samples were 4.5 ng/mL and 2.0 to 8.8 ng/mL, respectively. Mo levels in Japanese formula milk were 2 to 3 ng/mL. Based on the Mo levels in human milk and formula milk, the Mo intake of Japanese infants was estimated to be 2 to 4 microg/d/capita. PMID- 15895516 TI - Seasonal changes in the relationship between serum concentration of beta cryptoxanthin and serum lipid levels. AB - Beta-cryptoxanthin (beta-CRX) is a carotenoid pigment found especially in Satsuma mandarin (Citrus unshiu Marc.) fruit, which is mainly produced in Japan. Previously, we found that serum beta-CRX concentrations rose to extreme levels as the frequency of consumption of Satsuma mandarin increases. Using 94 non-smoking female volunteers, the present study evaluated the relationship between serum concentration of beta-CRX and serum lipid levels in September, when the Satsuma mandarin is not in season, and in January of the next year, when it is in season. The mean of increment in serum beta-CRX concentrations from September to January was 95.9 +/- 84.0 microg/dl (mean +/- SD). Although the changes in serum lipid levels did not correlate with the changes in serum beta-CRX, the serum HDL cholesterol and apo-lipoprotein A1 levels in the highest quartile of the increment in serum beta-CRX from September to January were significantly higher than those in the lowest quartile in both September and January. In cross sectional analyses, serum beta-CRX concentrations were correlated positively with those of LDL-cholesterol and apo-lipoprotein B levels in September, but these correlations were not observed in January. In contrast, serum beta-CRX concentrations were correlated positively with those of HDL-cholesterol and apo lipoprotein A1 levels in January. These results suggest that habitual eating of Satsuma mandarin while it is in season may influence lipid metabolism throughout the year. PMID- 15895517 TI - Conjugated linoleic acid deteriorates insulin resistance in obese/diabetic mice in association with decreased production of adiponectin and leptin. AB - Dietary supplementation of conjugated linoleic acids (CLA) is known to have some beneficial effects such as anti-carcinogenic and anti-obesity effects in several animal species, while it also induces insulin resistance and fatty liver, especially in mice. To explore the possible factors responsible for the CLA induced insulin resistance, we examined the plasma and mRNA expression levels of several adipocytokines, which are likely involved in the regulation of insulin sensitivity, in normal C5 7BL, mildly obese/diabetic KK and morbidly obese/diabetic KKAy mice. Feeding a diet supplemented with 0.5%, CLA oil consisting of 30.5/% c9, t11-CLA and 28.9% t10, c12-CLA for 4 wk resulted in a decrease in white adipose tissue (WAT), an increase in liver weight with excess accumulation of triglyceride, and insulin resistance associated with hyperglycemia and hyperinsulinemia. The plasma and WAT mRNA levels of leptin were higher in KK and KKAy mice than C57BI. mice, whereas those of adiponectin were higher in C5 7BL mice. CLA-feeding decreased the levels of leptin, adiponectin and resistin, especially in KK and KKAy mice. In contrast, tumor necrosis factor alpha (TNFalpha) mRNA levels were higher in KK and KKAy mice than C57BL mice, and were increased by CLA feeding. The present results thus indicate that CLA feeding promotes insulin resistance in obese/diabetic mice by at least inverse regulation of leptin and adiponectin, and TNFalpha, adipocytokines known to either ameliorate or deteriorate insulin sensitivity, respectively. PMID- 15895518 TI - Effects of a fermented vegetable product on fat deposition and bone metabolism in ovariectomized rats. AB - We examined the effects of a fermented vegetable product (FVP), fermented by yeast as well as lactic acid bacteria, on body weight and fat deposition after ovariectomy in 10-wk-old Sprague-Dawley rats. The rats were divided into the following 5 groups: (1) ovari-ectomized rats fed as usual (OVX); (2) ones receiving FVP at a daily dose of 100 mg/kg (LOW); (3) ones receiving FVP at a daily dose of 1000 mg/kg (HIGH); (4) ones receiving a calcium-free diet (Ca FREE); (5) sham-operated rats (SHAM). A slight increase in body weight was observed in the LOW group compared with the OVA group. The rate of increase in total fat content was lowest in the LOW group, but both bone weight and strength were similar to those in the other ovariectomized groups. In conclusion, the low dose of FVP turned out to reduce fat content without affecting bone weight and strength. PMID- 15895519 TI - Hydrophilic antimutagens in fermented defatted soybeans with Neurospora intermedia (D-oncom). AB - The author has previously reported on the higher antioxidative activity in vivo of defatted soybean oncom (D-oncom), fermented defatted soybeans with Neurospora intermedia, in comparison with that of defatted soybeans (DSB). In this paper, the hydrophilic antimutagenicity of D-oncom against N-nitrosodimethylamine (NDMA) was investigated. Water-extract of D-oncom had a stronger antimutagenicity than that of DSB. The main antimutagenic fraction of D-oncom was anionic, and stable against heating at 37 degrees C. Its antimutagenicity was about one sixth of that of ascorbic acid. Actually the fraction scavenged superoxide anions. Therefore, the antimutagenicity of D-oncom against NDMA might be involved in the reduction of oxidative stress by scavenging superoxide anions. Nonetheless the main antimutagenic fraction was not polyphenol, peptide nor nucleotide, its molecular weight being distributed about 5000-12,500. PMID- 15895520 TI - Effects of seasonings on the stability of ascorbic acid in a cooking model system. AB - The thermolability of ascorbic acid (AA) in aqueous solution at 100 degrees C was assessed in the presence of various seasonings commonly used in Japanese-style cooking. A model system approximated Japanese cooking with regard to the concentrations of AA and seasonings and the heating time. The decrease of AA in the reaction system of this experiment was a first-order reaction with respect to the concentration of AA loss. Although kinetic constants for AA loss decreased with increasing concentrations of AA (25-400 microg/mL), the absolute amounts degraded were almost the same for all AA concentrations, suggesting that dissolved oxygen is one of main factors affecting the stability of AA solutions during heating at 100 degrees C. When each seasoning was added to AA solution, salt stabilized AA and Japanese alcohol-containing admixtures, such as sake and sweet sake (mirin), did not have a significant effect on the stability. Conversely, soy sauce, miso (fermented soybean paste) and broth powder from skipjack accelerated the decrease of AA in a concentration-dependent manner. The kinetic study suggested that oxygen was rapidly consumed and AA loss accelerated by addition of soy sauce or miso to AA solution. Consequently it is likely that a reaction mechanism shifts from aerobic to anaerobic and the forward reactions proceed. Of the constituents of Japanese seasonings, not only iron but also amino acids are involved in the acceleration of AA degradation. The presence of amino acids should be taken into account when considering the levels of AA in soups. PMID- 15895521 TI - Occurrence of vitamin B12 in green, blue, red, and black tea leaves. AB - Vitamin B12 contents of green (0.046-0.263 and 0.125-0.535 microg/100 g dry weight), blue (0.068-0.081 and 0.525-0.528 microg/l00 g dry weight), red (0.061 and 0.663 microg/100 g dry weight), and black (0.104-0.859 and 0.305-1.20 microg/100 dry weight) tea leaves were obtained by intrinsic factor chemiluminescence and microbiological methods, respectively. Although vitamin B12 was found in all tea leaves tested by both assay methods, the higher values by the microbiological method were not due to occurrence of both deoxyribosides and deoxynucleotides (known as an alkali-resistant factor), but may have been due to that of inactive corrinoid compounds for mammals in the tea leaves. PMID- 15895522 TI - The difference between the basal metabolic rate and the sleeping metabolic rate in Japanese. AB - Previous studies have demonstrated the difference between the basal metabolic rate (BMR) and the sleeping metabolic rate (SMR): however, the difference in the Japanese population has not yet been explored. This study examined the relationship between the BMR and SMR in ninety-four healthy Japanese subjects (37 males and 57 females, 39 +/- 12 y of age and 22.0 +/- 7.4% body fat) in a respiratory chamber. The SMR was significantly lower than the BMR (1416 +/- 245 vs. 1492 +/- 256 kcal/d): however, there was a highly significant correlation between the two (r = 0.867; p < 0.001). The ratio of SMR/BMR largely varied among individuals (0.95 +/-0.08, 8.4% of the coefficient of variation). The ratio was significantly lower in males than in females (0.93 +/- 0.10 vs. 0.97 +/- 0.06, p < 0.05). None of the anthropometric measures (age, weight, body mass index, body surface area or percent body fat) correlated with the ratio. These results showed that SMR was 95%, of BMR on average in a healthy Japanese group. However, when applied over a longer time period (24 h or more), the difference tends to become negligible for most analyses in a group. Although the difference between SMR and BMR will induce a 5% gap of physical activity level defined as the total energy expenditure divided by the BMR or SMR, this factor seems to have little practical importance in epidemiological research. PMID- 15895523 TI - Lymphatic absorption of structured triacylglycerols, 1(3)-stearoyl-2,3 (1) dilinoleoylglycerol and 2-linoleoyl-1,3-distearoylglycerol in rats. AB - Lymphatic recovery of 1(3)-stearoyl-2,3(1)-dilinoleoylglycerol (SLL) and 2 linoleoyl-1,3-distearoylglycerol (SLS) which had been enzymatically synthesized were compared with those of trilinoleoylglycerol (LLL) and the randomly esterified triacylglycerol which contained stearic acid and linoleic acid at 1:2. Recoveries of linoleic acid in all of the triacylglycerols were more than 94.0%. Lymphatic 24 h-recoveries of stearic acid given as SLL and SLS were significantly lower than that of stearic acid given as the randomly esterified triacylglycerol. Recoveries of stearic acid from SLL, SLS and the randomly esterified triacylglycerol were 88.49%, 68.3% and 101%. respectively. PMID- 15895524 TI - 15-amido erythromycins: synthesis and in vitro activity of a new class of macrolide antibiotics. AB - An array of 15-amido substituted erythromycin A compounds was synthesized using a chemobiosynthesis approach. It was found that while the in vitro antibacterial activities of aryl amides were inferior to erythromycin A, substituted benzylamides showed equivalent and in some cases improved activity against the macrolide-resistant strains. The 15-amidoerythromycins represent a new class of antibacterial macrolides. PMID- 15895525 TI - Generation of new epothilones by genetic engineering of a polyketide synthase in Myxococcus xanthus. AB - Epothilones, potent cytotoxic agents and potential anticancer drugs, are complex polyketides produced by a modular polyketide synthase (PKS). The epothilone PKS genes were introduced and expressed in Myxococcus xanthus and engineered to generate novel unnatural natural products which can be used as new scaffolds for chemical modification. Inactivation of the KR domain in module 6 of the epo PKS resulted in accumulation of 9-oxoepothilone D and its isomer 8-epi-9 oxoepothilone D as the major products. Modification of the KR domain in module 4 resulted in the production of the expected compound 12,13-dihydro-13 oxoepothilone C in trace amounts, and the unexpected compound 11,12-dehydro-12,13 dihydro-13-oxoepothilone D as the major product. The other expected compound, 12,13-dihydro-13-oxoepothilone D, was not detected. The unexpected 13-oxo derivative produced indicates that the ER domain of module 5 has substrate specificity requirements and suggests a second enzymatic role for the domain. PMID- 15895526 TI - Absolute stereostructures of cell-adhesion inhibitors, peribysins A, E, F and G, produced by a sea hare-derived Periconia sp. AB - Peribysins E-G (1-3) have been isolated from a strain of Periconia byssoides originally separated from the sea hare Aplysia kurodai. Their absolute stereostructures have been elucidated on the basis of spectroscopic analyses using 1D and 2D NMR techniques and some chemical transformations. In addition, the absolute configuration of peribysin A (4), previously undetermined, has been established by conversion to 2 and 3. All these metabolites inhibited the adhesion of human-leukemia HL-60 cells to HUVEC. PMID- 15895527 TI - New antibiotic Sch 725424 and its dehydration product Sch 725428 from Kitasatospora sp. AB - A new microbial metabolite Sch 725424 (1) was isolated from the culture of Kitasatospora sp. The structure elucidation of 1 was accomplished based on NMR spectroscopic analyses as well as extensive structure elucidation of its dehydration product Sch 725428 (2). Compound 1 showed inhibitory activity against Staphylococcus aureus with MIC values 1-2 microg/ml, and also displayed weak antifungal activity against Saccharomyces cerevisiae (PM503) with an MIC 32 microg/ml. PMID- 15895528 TI - Identification and antibacterial activity of a new oleandomycin derivative from Streptomyces antibioticus. AB - During the study on the oleandomycin production, we purified a new oleandomycin derivative having a macrolactone of which biosynthesis does not follow the genetic architecture of the oleandomycin PKS. The molecular formula for the compound was suggested as C35H59NO11 on the basis of the analysis of NMR and HRMS data (m/z 670.4185, Delta-1.9mmu, calcd for C35H60NO11). 13C NMR assignments and analysis of COSY, HMBC and HMQC data suggested that the compound differs from oleandomycin by formation of the olefinic functionality resulting from the dehydration of a hydroxy group in oleandomycin. The new oleandomycin derivative has antibacterial activities similar to those of oleandomycin agaisnt Enterococcus faecalis, Bacillus subtilis and Staphylococcus aureus. PMID- 15895529 TI - Effect of cytogenin, a novel immunomodulator, on streptozotocin-induced diabetes in mice. AB - The anti-diabetic effect of cytogenin was examined using streptozotocin-induced diabetes in mice. Cytogenin suppressed not only the increase of plasma glucose level but also the body weight reduction in diabetic mice. Histological examination of the pancreas taken from diabetic mice given cytogenin showed that cytogenin decreased the number of macrophages infiltrated into islet of pancreas. Further, cytogenin suppressed the nitric oxide generation by macrophages treated with lipopolysaccharide through decreasing of inducible nitric oxide synthase expression. Cytogenin suppressed interleukin-6 expression by macrophage treated with LPS, suggesting that the anti-diabetic activity of cytogenin might be partly attributed to the suppressive activity against nitric oxide generation. PMID- 15895530 TI - Prunustatin A, a novel GRP78 molecular chaperone down-regulator isolated from Streptomyces violaceoniger. AB - In the course of our screening program for regulators of a molecular chaperone GRP78 expression, we isolated a novel inhibitor of GRP78 expression, designated as prunustatin A, from Streptomyces violaceoniger 4521-SVS3. The structure of prunustatin A was determined by a series of NMR analyses to be an oxidized type of the neoantimycin family. Prunustatin A inhibited the expression of GRP78 induced by 2-deoxyglucose in human fibrosarcoma HT1080 cells accompanied by global cell death without showing cytotoxicity under normal nutrient condition. PMID- 15895531 TI - Deoxyverrucosidin, a novel GRP78/BiP down-regulator, produced by Penicillium sp. AB - Glucose-regulated protein 78 (GRP78) resides in endoplasmic reticulum (ER) and plays a role in protecting tumor cells against the toxic effects of anticancer agents. During the search for down-regulators of GRP78 using a reporter gene (luciferase) assay system, we isolated a novel compound designated as deoxyverrucosidin (1), a congener of verrucosidin (2), from Penicillium sp. and identified it as a down-regulator of the grp78 gene. The structure of 1 was determined by mainly ESI-mass and two-dimensional NMR spectra. 1 dose-dependently inhibited the expression of GRP78 promoter with an IC50 of 30 nM. PMID- 15895532 TI - What works best? Knowledge, skills, motivation, and opportunity. PMID- 15895533 TI - The conspicuous absence of people with disabilities in public fitness and recreation facilities: lack of interest or lack of access? AB - Critical Issues and Trends More than 50 million Americans have a disability. These people face enormous physical, social, and attitudinal barriers toward their participation in physical and recreational activities that they need to maintain their health and wellness. Furthermore, the concept of being "healthy" and "disabled" or "physically active" and "disabled" is not a common visualization in the mainstream media. This paper calls for a more inclusive vision within public health messages that target physical activity participation among its citizens, especially those with disability who are at greater risk for developing health problems associated with sedentary living. PMID- 15895534 TI - Healthy nutrition environments: concepts and measures. AB - The authors provide a conceptual model of a healthy nutrition environment, then review the types of measures required to assess various aspects of this environment. Measures fall into priority categories of consumer and community environments. PMID- 15895535 TI - Use of interactive health communication to affect smoking intentions in middle school students: a pilot test of the "Headbutt" risk assessment program. AB - PURPOSE: Developing and disseminating innovative and effective approaches for smoking prevention among middle school children remains a public health priority. This pilot study evaluates the use of a Web-based tobacco prevention program, Headbutt, to change intentions of middle school children to smoke tobacco. METHODS: Headbutt was implemented with the use of a single-group pretest-posttest study design in sixth grade classes of nine middle schools in Texas (student n = 2227). The program assesses cognitive determinants of smoking and provides intervention feedback tailored to the child's responses. RESULTS: Headbutt significantly affected smoking intentions, prosmoking attitudes, self-efficacy expectations, and knowledge of negative consequences (all p < or = .001) measured with scales adopted from the Texas Tobacco Initiative Survey. Change in prosmoking attitudes had the greatest predictive effect on smoking intentions (p < .001). These results were moderated by ethnicity and age of students. CONCLUSION: Findings need to be interpreted in the light of study design limitations. However, strong associations between the Headbutt program and intention change suggests that a more rigorous effectiveness trial is indicated. PMID- 15895536 TI - Smoking cessation and exercise promotion counseling in psychologists who practice psychotherapy. AB - PURPOSE: Little is known about health promotion within the context of psychotherapy. The present study assessed a sample of psychologists' attitudes and behavior with their psychotherapy clients regarding smoking cessation and exercise promotion counseling. DESIGN: This is a cross-sectional survey study. SETTING: Surveys were mailed. SUBJECTS: Licensed psychologists (1000) in Massachusetts were randomly selected to receive surveys. Psychologists practicing psychotherapy at least 5 h/wk were invited to participate. Out of 496 responses, 328 completed surveys were analyzed. MEASURES: Two parallel surveys were developed for smoking (N = 154) and exercise (N = 174) assessing health promotion behaviors and attitudes. RESULTS: Approximately 43% of respondents inquire about smoking and 53% inquire about exercise with new clients. Over 45% advise smoking clients to quit and 50% advise sedentary clients to exercise. Over 80% of respondents felt smoking and exercise should be addressed in psychotherapy; however only 41% of smoking and 65% of exercise respondents felt confident in their counseling abilities. Lack of confidence, beliefs about relevance of smoking and exercise to psychological functioning, and beliefs that such personal choices should not be addressed in psychotherapy significantly predicted smoking cessation and exercise promotion behaviors. CONCLUSION: Many respondents are engaging in health promotion with their psychotherapy clients. Exercise promotion is occurring more frequently and is viewed more favorably compared with smoking cessation counseling. Health promotion attitudes are associated with counseling behaviors. Limitations include self-selection bias and small sample size. PMID- 15895537 TI - Intention to quit smoking, attempts to quit, and successful quitting among Hong Kong Chinese smokers: population prevalence and predictors. AB - PURPOSE: To assess the prevalence of each step in the smoking-cessation process (intention to quit, attempts to quit, and successful quitting) and to examine the factors associated with them among Chinese smokers. DESIGN: A cross-sectional survey of subjects from randomly selected households. SETTING: Four thousand one hundred forty-two households in Hong Kong. SUBJECTS: A total of 11,779 persons, aged 15 years or older, were enumerated (response rate = 74.0%). MEASURES: A validated structured questionnaire was used for data collection. The questionnaire sought information on the subject's sociodemographic background, smoking habits, and workplace attitude to smoking. The predictors for successful quitting, past quitting attempts, and intention to quit were assessed by chi2 tests and multiple logistic regression. RESULTS: Of the respondents, 14.4% were current smokers, 7.5% were ex-smokers, and 78.1% were nonsmokers. Of the daily smokers, 52% intended to quit. The factors associated with quitting were being married, being in the student/retired/others category, being older, having received higher education, not smoking to kill time, and smoking because of curiosity. Being married and not smoking to kill time were associated with past quitting attempts. Being male, married, and not smoking to kill time were associated with the intention to quit smoking. CONCLUSION: The findings of this study indicate that differing predictors may contribute to the different transitional stages of smoking cessation. Population-based smoking-cessation programs should take these predictors into consideration in the design of interventions. PMID- 15895538 TI - Facilitating adolescent smoking: who provides the cigarettes? AB - PURPOSE: Most adolescent smokers obtain cigarettes through social sources. We examine the extent to which cigarettes are provided by facilitators of legal age to purchase cigarettes. DESIGN: Analyses of data from the 1999 California Tobacco Survey, a large population-based, random-digit-dialed telephone survey, are reported. SETTING: California. SUBJECTS: Data were from a subset of 1239 adolescent (12-17 years) respondents who reported ever having smoked a cigarette. The response rate for all adolescents selected for interview was 75.5%. MEASURES: We describe cigarette providers to adolescents in social (cigarettes given to the adolescent) and economic (someone else buys cigarettes for the adolescent) transactions by the reported facilitator's age. RESULTS: Of the 82.2% +/- 2.6% of adolescents who had ever smoked who usually obtained cigarettes from others, 21.6% +/- 2.5% used economic transactions; most (60.6% +/- 3.4%) were given cigarettes. The majority (73.3% +/- 3.6%) of those relying on social sources were given cigarettes by someone < 18 years of age; very few were given cigarettes by someone 21+ years old. Most (90.4% +/- 2.0%) usually given cigarettes reported friends as facilitators. Of those who relied on economic transactions, 56.1% +/- 6.6% reported facilitators who were 18- to 20-year-olds, another 24.7% +/- 6.3% had suppliers > or = 21 years of age. Altogether, 80.8% +/- 5.8% of facilitators in economic transactions were > or = 18 years of age. CONCLUSIONS: Until peer approval of smoking and sharing cigarettes and adult facilitation of adolescent smoking is reduced, it will be difficult to significantly reduce adolescents' access to cigarettes. PMID- 15895539 TI - Textual analysis of tobacco editorials: how are Key media gatekeepers framing the issues? AB - PURPOSE: The news media's potential to promote awareness of health issues is established, and media advocacy is now an important tool in combating tobacco use. This study examines newspaper editors' perspectives of tobacco-related issues. DESIGN: This study presents a textual analysis of tobacco-related editorials. The data consist of editorials on tobacco from a sample of 310 U.S. daily newspapers over the course of 1 year (2001). Data were sampled from a random one-third of the days per month, yielding 162 editorials for analysis. A qualitative textual analysis was conducted. Each editorial was coded for theme, position, and frame. We analyzed the topics gaining editorial attention and the arguments made to support various perspectives. RESULTS: Editorials discussed a variety of both positive and negative news events, largely conveying support for tobacco-control objectives. Various organizing frames were used-supporting policy interventions, condemning the industry, highlighting individual rights, and expressing general cynicism were most prevalent. CONCLUSION: Editors largely promoted tobacco-control efforts, particularly policy advances. There was, however, little coverage of key issues such as health effects and addiction perhaps because they are no longer perceived to be contentious. Advocates should seek to address this area and minimize the cynicism of key media gatekeepers to avoid undermining policy and individual change efforts. PMID- 15895540 TI - Characteristics of adults who use prayer as an alternative therapy. AB - PURPOSE: To describe the demographics, health-related and preventive-health behaviors, health status, and health care charges of adults who do and do not pray for health. DESIGN: Cross-sectional survey with 1-year follow-up. SETTING: A Minnesota health plan. SUBJECTS: A stratified random sample of 5107 members age 40 and over with analysis based on 4404 survey respondents (86%). MEASURES: Survey data included health risks, health practices, use of preventive health services, satisfaction with care, and use of alternative therapies. Health care charges were obtained from administrative data. RESULTS: Overall, 47.2% of study subjects reported that they pray for health, and 90.3% of these believed prayer improved their health. After adjustment for demographics, those who pray had significantly less smoking and alcohol use and more preventive care visits, influenza immunizations, vegetable intake, satisfaction with care, and social support and were more likely to have a regular primary care provider. Rates of functional impairment, depressive symptoms, chronic diseases, and total health care charges were not related to prayer CONCLUSIONS: Those who pray had more favorable health-related behaviors, preventive service use, and satisfaction with care. Discussion of prayer could help guide customization of clinical care. Research that examines the effect of prayer on health status should adjust for variables related both to use of prayer and to health status. PMID- 15895541 TI - Health promotion referrals in an urban clinic: removing financial barriers influences physician but not patient behavior. AB - PURPOSE: This study describes the prevalence and characteristics of physician health promotion referrals and patient adherence to referrals in a community based primary care clinic and associated wellness facility. The role of reimbursement for attendance to the wellness facility was specifically examined. DESIGN: Retrospective cohort study. SETTING: The Church Health Center of Memphis, Tennessee: a low-income urban clinic and its affiliated wellness center. SUBJECTS: Patients were primarily African-American, lower-income, urban residents of Shelby County, Tennessee. MEASURES: All study data came from existing medical clinic and wellness facility records of utilization, patient history, and diagnoses. RESULTS: Of 6321 clinic patients, 16.7% (n = 1069) received a provider health promotion referral. Logistic regression analyses identified that physician referral was related to patient factors of access to free wellness-facility membership, employment status, receiving a behavior-related diagnosis, and being African-American and female. Of patients receiving a referral 17.2% (n = 184) adhered to this advice and visited the wellness facility. New patients were more likely to adhere to a referral than established patients. CONCLUSION: Demographic, financial, and patient characteristics influenced whether health behavior change referrals were made by primary care physicians in a community clinic. Removing financial barriers did not influence patient adherence, but new patients were more likely to follow the recommendation than those previously seen at the clinic. PMID- 15895542 TI - City and regional planning: a primer for public health officials. AB - Recognizing that planners' decisions affect the public's health, some public health officials are becoming more involved in city and regional planning. This article describes city and regional planning fundamentals to help public health practitioners better understand plan making and plan implementation, including the development project review process; provides examples of how three local public health agencies are currently involved in planning; and discusses general strategies for such participation. With this information, public health officials could increase their influence on local planning with consequent public health benefits. PMID- 15895543 TI - On questions no one can answer. PMID- 15895544 TI - End-of-life care in a children's hospice program. AB - A project was completed in 1999 to evaluate the Canuck Place children's hospice program. This article reports only on the end-of-life care component. Results are provided from mail-out surveys with families who required end-of-life care and are supplemented by qualitative data which were collected from interviews with individuals prior to the survey. Eighteen families completed face-to-face interviews and another 70 families completed the mail-out questionnaire developed from the initial interviews. A total of 39 parents responded to the survey section about end-of-life care services. Results indicated that parents were reasonably comfortable discussing death with staff; they generally felt well prepared for their child's death because of the staff; the death of another child had a significant effect on families; parents' wishes at the time of their child's death were very supported by staff; and families were well supported by staff at the time of a child's death. Implications for practice and suggestions for future research are discussed. PMID- 15895545 TI - Educational needs of palliative care nurses in Quebec. AB - Education appears to be a useful strategy for improving quality of care and work related stress management. The present study assesses the educational needs of palliative care nurses working in acute care hospitals (n=88) and CLSCs (Centre Local de Services Communautaires, home-based, n=109) of the larger Quebec City area. It also describes relationships between educational needs, psychological distress, and self-efficacy. Nurses were surveyed and compared on training needs, preferred learning format, as well as barriers and factors facilitating attendance to workshops. Similar educational needs were observed in both work settings. Crisis, stress, and emotion management were rated as the most useful topics, followed by nonpharmacological ways to manage pain. Preferred educational formats were lectures and group discussions. Attending free training sessions during work hours was described as the best way to promote participation. Educational needs were positively related to psychological distress, and negatively related to perceived self-efficacy in providing good palliative care. PMID- 15895546 TI - Reaching common ground: a patient-family-based conceptual framework of quality EOL care. AB - Improvement in the quality of end-of-life (EOL) care is a priority health care issue since serious deficiencies in quality of care have been reported across care settings. Increasing pressure is now focused on Canadian health care organizations to be accountable for the quality of palliative and EOL care delivered. Numerous domains of quality EOL care upon which to create accountability frameworks are now published, with some derived from the patient/family perspective. There is a need to reach common ground on the domains of quality EOL care valued by patients and families in order to develop consistent performance measures and set priorities for health care improvement. This paper describes a meta-synthesis study to develop a common conceptual framework of quality EOL care integrating attributes of quality valued by patients and their families. PMID- 15895547 TI - The French version of the palliative care quiz for nursing (PCQN-F): development and evaluation. AB - This article describes the process and outcomes of translating the Palliative Care Quiz for Nursing into French (PCQN-F). A process of decentering, translation/back-translation, and review served to ensure grammatical, structural, and conceptual equivalence with the English version of the quiz. A total of 189 nursing personnel participated in the evaluation. Evaluation involved determining overall responses to the quiz, item to total correlations, reliability, mean score comparisons, and the most frequent misconceptions about palliative care nursing. Our initial review demonstrates the face and content validity of the PCQN-F. Further testing is encouraged to more fully demonstrate its psychometric and educational properties. PMID- 15895548 TI - Navigating uncharted territory: experiences of families when a child is dying. AB - In this grounded theory study, eight families moved through a process of navigating uncharted territory as they lived with their child who was dying from a neurodegenerative illness. The emotions of fear, uncertainty, and grief gave impetus to the process. Families lived much of their lives on plateaus of relative stability, where they often felt alone and isolated from health care professionals. Periods of instability led to families dropping off the plateau. Living with a dying child had profound physical, emotional, and financial effects on families. The process was characterized by four dimensions. Parents used strategies within these dimensions to manage the physical, cognitive, and emotional work arising from the situation. These dimensions and strategies were constrained and facilitated by four intervening conditions. In addition, the process occurred within the broader context of the acute, curative health care system and the sociocultural environment. Suggestions for practice and research are offered to assist health care professionals in providing optimal care to these families. PMID- 15895549 TI - Evaluation of education in palliative care: determining the effects on nurses' knowledge and attitudes. AB - Our study was an evaluation of a pilot course in palliative nursing care designed to improve practising nurses' knowledge regarding cancer pain management and attitudes toward care of the dying patient. The course involved 12 weeks (36 hours) of systematic classroom instruction for registered nurses, and a one-day observational experience on a designated palliative care unit. A total of 16 nurses participated in the study. A repeated measures design was used to evaluate the effectiveness of the course. Attitudes toward care of the dying patient and his/her family were also evaluated. Results showed that nurses' attitudes improved significantly from Time 1 (first day of course) to Time 2 (last day of course) (p=0.0007), and that this improvement was maintained at Time 3 (three months later) (p=0.064). The observational experience on a palliative care unit held in conjunction with the course was reported to be helpful to students in consolidating theoretical instruction. PMID- 15895550 TI - Continuity of care for advanced cancer patients. PMID- 15895551 TI - Confession at the end of life: a Jewish perspective. PMID- 15895552 TI - The wide spectrum of spinocerebellar ataxias (SCAs). AB - Spinocerebellar ataxias (SCAs) are a clinically heterogeneous group of disorders. Current molecular classification corresponds to the order of gene description (SCA1-SCA 25). The prevalence of SCAs is estimated to be 1-4/100,000. Patients exhibit usually a slowly progressive cerebellar syndrome with various combinations of oculomotor disorders, dysarthria, dysmetria/kinetic tremor, and/or ataxic gait. They can present also with pigmentary retinopathy, extrapyramidal movement disorders (parkinsonism, dyskinesias, dystonia, chorea), pyramidal signs, cortical symptoms (seizures, cognitive impairment/behavioral symptoms), peripheral neuropathy. SCAs are also genetically heterogeneous and the clinical diagnosis of subtypes of SCAs is complicated by the salient overlap of the phenotypes between genetic subtypes. The following clinical features have some specific values for predicting a gene defect: slowing of saccades in SCA2, ophthalmoplegia in SCA1, SCA2 and SCA3, pigmentary retinopathy in SCA7, spasticity in SCA3, dyskinesias associated with a mutation in the fibroblast growth factor 14 (FGF 14) gene, cognitive impairment/behavioral symptoms in SCA17 and DRPLA, seizures in SCA10, SCA17 and DRPLA, peripheral neuropathy in SCA1, SCA2, SCA3, SCA4, SCA8, SCA18 and SCA25. Neurophysiological findings are compatible with a dying-back axonopathy and/or a neuronopathy. Three patterns of atrophy can be identified on brain MRI: a pure cerebellar atrophy, a pattern of olivopontocerebellar atrophy, and a pattern of global brain atrophy. A remarkable observation is the presence of dentate nuclei calcifications in SCA20, resulting in a low signal on brain MRI sequences. Several identified mutations correspond to expansions of repeated trinucleotides (CAG repeats in SCA1, SCA2, SCA3, SCA6, SCA7, SCA17 and DRPLA, CTG repeats in SCA8). A pentanucleotide repeat expansion (ATTCT) is associated with SCA10. Missense mutations have also been found recently. Anticipation is a main feature of SCAs, due to instability of expanded alleles. Anticipation may be particularly prominent in SCA7. It is estimated that extensive genetic testing leads to the identification of the causative gene in about 60-75 % of cases. Our knowledge of the molecular mechanisms of SCAs is rapidly growing, and the development of relevant animal models of SCAs is bringing hope for effective therapies in human. PMID- 15895553 TI - The Anp32 family of proteins containing leucine-rich repeats. AB - Herein we describe the characteristic features of the Anp32 family represented by the cerebellar leucine-rich repeat protein (Lanp) and the cerebellar developmental-regulated protein 1 (Cpd1). The Anp32 family consists of 32 evolutionarily-conserved proteins and is included within the superfamily of leucine-rich repeat (LRR) proteins characterized by the presence of tandem arrays of a LRR, a structural motif implicated in the mediation of protein-protein interactions. We describe three novel human Anp32 proteins, reveal the evolutionary relationships of the members of the Anp32 family, provide insights into their biochemical and structural properties, and review their macromolecular interactions, substrate specificities, tissue distribution/expression patterns, and physiological and pathological roles. Recent findings indicate a conserved role of members of the Anp32 family during evolution in the modulation of cell signalling and transduction of gene expression to regulate the morphology and dynamics of the cytoskeleton, cell adhesion, neural development or cerebellar morphogenesis. PMID- 15895554 TI - Pathogenesis of spinocerebellar ataxias viewed from the RNA perspective. AB - The underlying cause of a number of autosomal dominant spinocerebellar ataxias is the expansion of various types of simple sequence repeats located in diverse functional regions of different single genes. The genetic heterogeneity of these diseases which contrasts with the similarity of their pathology sites and clinical symptoms justifies the search for the shared mechanism of pathogenesis. In this article we discuss the arguments which are in favor of the RNA-mediated pathomechanism. PMID- 15895555 TI - Embryonic stem cell models of CAG repeat disease. AB - Nine neurodegenerative disorders are caused by CAG/polyglutamine (polyQ) repeat expansions. The underlying molecular mechanisms responsible for disease specific neurodegeneration remain elusive. In vivo and in vitro models utilizing rodent tissues, immortalized human cell lines, and human post mortem samples have provided insight into disease mechanisms. Concern that cellular and molecular processes observed in these models may not faithfully reproduce human diseases or be useful to identify compounds of therapeutic utility has driven development of new disease models. In addition to their therapeutic potential, stem cells represent a renewable source of tissue that can be directed into neurons and glia and can be used to study neurodegenerative cascades from their inception. Neuronally differentiated human stem cells containing CAG repeat expansions have the potential to accurately replicate human CAG repeat diseases and may be a faithful predictor of which compounds will be of human benefit. As a first step in development of this type of model, we developed murine embryonic stem cell models to study the mechanisms of polyQ tract induced neuronal degeneration. PMID- 15895556 TI - HSP27 and cell death in spinocerebellar ataxia type 3. AB - Spinocerebellar ataxia type 3 (SCA3) is an autosomal dominant spinocerebellar degeneration characterized by a wide range of clinical manifestations. In this review, we discuss the role(s) that heat shock protein 27 (HSP27) may play in the cell death process of spinocerebellar ataxia type 3. PMID- 15895557 TI - Recent progress in spinocerebellar ataxia type-10 (SCA10). AB - Spinocerebellar ataxia type 10 (SCA10) is a dominantly inherited ataxia caused by expansion of ATTCT pentanucleotide repeat in intron 9 of a novel gene, E46L, on chromosome 22q13.3. SCA10 is a complex neurodegenerative condition. Initial studies characterized SCA10 as pure cerebellar ataxia associated with seizures. Recent identification of new SCA10 families revealed more diverse phenotypes, including polyneuropathy, pyramidal signs, cognitive and neuropsychiatric impairment. Moreover, several families manifest with ataxia without seizures. Thus a complete clinical spectrum is emerging. Progress has also been made in understanding the molecular and genetic mechanisms of pathogenesis. The length of expanded ATTCT repeats is variable in different tissues and highly unstable during paternal transmission, revealing complex genetic and pathogenetic processes. Under torsional stress, ATTCT repeats form unpaired DNA structure and may serve as an erroneous DNA replication origin, potentially contributing to repeat instability and aberrant cell cycle entry. E46L is a cytoplasmic protein with unknown function. Reduced expression of E46L in primary neuronal cultures from cerebellum and cortex by small interfering RNAs (siRNAs) caused increased apoptosis, raising the possibility that reduced expression of E46L might also play an important role in SCA10 pathogenesis. PMID- 15895558 TI - Spinocerebellar ataxia with mental retardation (SCA13). AB - Spinocerebellar ataxia 13 is a slowly progressive and relatively pure autosomal dominant cerebellar ataxia with childhood onset and mental deficiency. The responsible gene has been assigned to a 5.2 Mbases interval on chromosome 19q in a single French family. PMID- 15895559 TI - Spinocerebellar ataxia type 15. AB - Spinocerebellar ataxia type 15 (SCA15) was first reported in 2001 on the basis of a single large Anglo-Celtic family from Australia, the locus mapping to chromosomal region 3p24.2-3pter. The characteristic clinical feature was of very slow progression, with two affected individuals remaining ambulant without aids after over 50 years of symptoms. Head and/or upper limb action tremor, and gaze evoked horizontal nystagmus were seen in several persons. MRI brain scans showed predominant vermal atrophy, sparing the brainstem. In 2004, a Japanese pedigree was reported, which displayed very similar clinical features to the original SCA15 family, and which mapped to an overlapping candidate region. These two families might plausibly reflect a locus homogeneity, but for the present this remains an open question. PMID- 15895560 TI - Clinical, psychological, and genetic characteristics of spinocerebellar ataxia type 19 (SCA19). AB - The SCA19 locus on chromosome 1p21-q21 was identified in a Dutch family in 2002. Affected individuals displayed a lateonset slowly progressive mild cerebellar ataxia, hyporeflexia, and signs of frontal lobe dysfunction. A postural head tremor and myoclonic movements were observed occasionally. Before the SCA19 locus was identified, the SCA22 symbol had been assigned to a locus on 1p21-q23 following a linkage study of a Chinese family with spinocerebellar ataxia. Although both SCA19 and SCA22 are linked to 1p21-q21, the clinical features are slightly different. While it cannot be excluded that the genes lie in close approximation at this locus, it is more likely that the same gene is mutated in both the Dutch and Chinese families, and that SCA19 and SCA22 represent the same condition. PMID- 15895561 TI - Spinocerebellar ataxia type 20. AB - Spinocerebellar ataxia type 20 (SCA20) was reported in 2004 in a single Australian Anglo-Celtic pedigree. The phenotype is distinctive, with palatal tremor, and hypermetric saccades, and early dentate (but not pallidal) calcification in the absence of abnormalities of calcium metabolism. Dysarthria, rather than gait ataxia, was the initial symptom in most, and was typically conjoined with dysphonia, clinically resembling adductor spasmodic dysphonia. The onset of these speech abnormalities was abrupt in some cases. MRI scanning showed mild to moderate pancerebellar atrophy with dentate calcification, with olivary pseudohypertrophy in some cases, in the absence of other brainstem or cerebral changes. Nerve conduction studies were normal. Progression appeared to be slow. SCA20 is probably rare, as despite the distinctive phenotype, only this one pedigree has been described. The locus mapped to the pericentromeric region of chromosome 11 with a LOD score of 4.47, and its candidate region overlaps that of SCA5. It seems probable that these two SCAs may be separate genetic entities, on the basis of their divergent clinical features, but formal proof awaits discovery of one or both responsible genes. PMID- 15895562 TI - Spinocerebellar ataxia with sensory neuropathy (SCA25). AB - Spinocerebellar ataxia 25 (SCA25) is a rare form of autosomal dominant cerebellar ataxia associated with a severe sensory neuropathy. Clinical variability ranges from incomplete penetrance at age 61 to a Friedreich ataxia-like syndrome. The responsible locus was mapped to chromosome 2p in a large region of 14 Mbases in a single French kindred. PMID- 15895563 TI - The pathogenesis of spinocerebellar ataxia. AB - Six forms of spinocerebellar ataxia (SCA) are caused by pathological cytosine adenine-guanine (CAG) trinucleotide repeat expansions in the coding region of the mutated genes. The translated proteins contain abnormally long polyglutamine stretches, and SCA-1, SCA-2, SCA-3/Machado-Joseph disease (MJD), SCA-6, SCA-7, and SCA-17 are "polyglutamine diseases". Despite their clinical and genetic heterogeneity, the ataxia-causing lesions in the brain invariably affect the "cerebellar module" that is defined as a reciprocal circuitry between the cerebellar cortex, the dentate nuclei, and the inferior olivary nuclei. While the neurons of the basis pontis do not properly belong to this module, pontine atrophy is an important additional lesion in SCA-1, SCA-2, and SCA-7. The descriptive term olivopontocerebellar atrophy (OPCA) applies to these forms whereas SCA-6 is the prototype of "pure" cerebellar cortical or cerebello-olivary atrophy. Purkinje cells have an elaborate dendritic tree, and atrophy of these most remarkable cells has captured the attention of many morphologists. Almost invariably, the loss of Purkinje cells entails retrograde neuronal degeneration in the inferior olivary nuclei. However, SCA-6 is an exception, and many olivary neurons survive. Similarly, stellate, basket, and granule cells do not undergo commensurate retrograde atrophy when Purkinje cells disappear. The dentate nucleus displays "grumose" degeneration in SCA-3/MJD while the cerebellar cortex and the inferior olivary nuclei remain largely unaffected. The role of polyglutamine-containing intranuclear and cytoplasmic inclusion bodies in SCA remains unknown but protein aggregation may be the common step in the pathogenesis of these otherwise rather heterogeneous disorders. PMID- 15895564 TI - Graduation from a CAMPEP or equivalent accredited graduate or residency program should be a prerequisite for certification in radiological physics. For the proposition. PMID- 15895565 TI - Graduation from a CAMPEP or equivalent accredited graduate or residency program should be a prerequisite for certification in radiological physics. Against the proposition. PMID- 15895566 TI - Retrospective analysis of 2D patient-specific IMRT verifications. AB - We performed 858 two-dimensional (2D) patient-specific intensity modulated radiotherapy verifications over a period of 18 months. Multifield, composite treatment plans were measured in phantom using calibrated Kodak EDR2 film and compared with the calculated dose extracted from two treatment planning systems. This research summarizes our findings using the normalized agreement test (NAT) index and the percent of pixels failing the gamma index as metrics to represent the agreement between measured and computed dose distributions. An in-house dose comparison software package was used to register and compare all verifications. We found it was important to use an automatic positioning algorithm to achieve maximum registration accuracy, and that our automatic algorithm agreed well with anticipated results from known phantom geometries. We also measured absolute dose for each case using an ion chamber. Because the computed distributions agreed with ion chamber measurements better than the EDR2 film doses, we normalized EDR2 data to the computed distributions. The distributions of both the NAT indices and the percentage of pixels failing the gamma index were found to be exponential distributions. We continue to use both the NAT index and percent of pixels failing gamma with 5%/3 mm criteria to evaluate future verifications, as these two metrics were found to be complementary. Our data showed that using 2%/2 mm or 3%/3 mm criteria produces results similar to those using 5%/3 mm criteria. Normalized comparisons that have a NAT index greater than 45 and/or more than 20% of the pixels failing gamma for 5%/3 mm criteria represent outliers from our clinical data set and require further analysis. Because our QA verification results were exponentially distributed, rather than a tight grouping of similar results, we continue to perform patient-specific QA in order to identify and correct outliers in our verifications. The data from this work could be useful as a reference for other clinics to indicate anticipated trends in 2D verifications under various conditions. PMID- 15895567 TI - Cardiac cone-beam CT volume reconstruction using ART. AB - Modern computed tomography systems allow volume imaging of the heart. Up to now, approximately two-dimensional (2D) and 3D algorithms based on filtered backprojection are used for the reconstruction. These algorithms become more sensitive to artifacts when the cone angle of the x-ray beam increases as it is the current trend of computed tomography (CT) technology. In this paper, we investigate the potential of iterative reconstruction based on the algebraic reconstruction technique (ART) for helical cardiac cone-beam CT. Iterative reconstruction has the advantages that it takes the cone angle into account exactly and that it can be combined with retrospective cardiac gating fairly easily. We introduce a modified ART algorithm for cardiac CT reconstruction. We apply it to clinical cardiac data from a 16-slice CT scanner and compare the images to those obtained with a current analytical reconstruction method. In a second part, we investigate the potential of iterative reconstruction for a large area detector with 256 slices. For the clinical cases, iterative reconstruction produces excellent images of diagnostic quality. For the large area detector, iterative reconstruction produces images superior to analytical reconstruction in terms of cone-beam artifacts. PMID- 15895568 TI - Patient self-attenuation and technologist dose in positron emission tomography. AB - Positron emission tomography (PET), with 511-keV radiation and long patient uptake times, presents unique radiation safety concerns. This two-part study considers aspects of PET radiation safety as they relate to PET suite design, dose to the public, and technologist occupational dose. In the first part of the study, the self-attenuation of radiation by patients' bodies was quantified. The radiation exposure was measured at three positions from 64 patients injected with fluorine-18 fluorodeoxyglucose (FDG) during the uptake period. Compared with an in vitro control used as a point source, a significant decrease in exposure (>40% at 1 m) was observed due to nonuniform distribution of FDG and attenuation within the patients. The attenuation data are consistent with results from simulations [M. E. Phelps, "Comments and Perspectives," J. Nucl. Med. 45, 1601 (2004)] that treat the body as a uniform, water-filled cylinder. As distance is often the principal source of protection for 511-keV radiation, the considerable self attenuation may allow for more compact PET suites. However, despite high patient self-attenuation, shielding, and standard precautionary measures, PET technologist occupational doses can remain quite high (approximately 12 mSv/year). The second part of this study tracked the daily dose received by PET technologists. Close technologist-patient interaction both during and following FDG administration, as much as 20 min/study, contribute to the high doses and point to the need for a more innovative approach to radiation protection for PET technologists. PMID- 15895569 TI - Effects of x-ray and CT image enhancements on the robustness and accuracy of a rigid 3D/2D image registration. AB - A rigid body three-dimensional/two-dimensional (3D/2D) registration method has been implemented using mutual information, gradient ascent, and 3D texturemap based digitally reconstructed radiographs. Nine combinations of commonly used x ray and computed tomography (CT) image enhancement methods, including window leveling, histogram equalization, and adaptive histogram equalization, were examined to assess their effects on accuracy and robustness of the registration method. From a set of experiments using an anthropomorphic chest phantom, we were able to draw several conclusions. First, the CT and x-ray preprocessing combination with the widest attraction range was the one that linearly stretched the histograms onto the entire display range on both CT and x-ray images. The average attraction ranges of this combination were 71.3 mm and 61.3 deg in the translation and rotation dimensions, respectively, and the average errors were 0.12 deg and 0.47 mm. Second, the combination of the CT image with tissue and bone information and the x-ray images with adaptive histogram equalization also showed subvoxel accuracy, especially the best in the translation dimensions. However, its attraction ranges were the smallest among the examined combinations (on average 36 mm and 19 deg). Last the bone-only information on the CT image did not show convergency property to the correct registration. PMID- 15895570 TI - Four-dimensional computed tomography: image formation and clinical protocol. AB - Respiratory motion can introduce significant errors in radiotherapy. Conventional CT scans as commonly used for treatment planning can include severe motion artifacts that result from interplay effects between the advancing scan plane and object motion. To explicitly include organ/target motion in treatment planning and delivery, time-resolved CT data acquisition (4D Computed Tomography) is needed. 4DCT can be accomplished by oversampled CT data acquisition at each slice. During several CT tube rotations projection data are collected in axial cine mode for the duration of the patient's respiratory cycle (plus the time needed for a full CT gantry rotation). Multiple images are then reconstructed per slice that are evenly distributed over the acquisition time. Each of these images represents a different anatomical state during a respiratory cycle. After data acquisition at one couch position is completed, x rays are turned off and the couch advances to begin data acquisition again until full coverage of the scan length has been obtained. Concurrent to CT data acquisition the patient's abdominal surface motion is recorded in precise temporal correlation. To obtain CT volumes at different respiratory states, reconstructed images are sorted into different spatio-temporally coherent volumes based on respiratory phase as obtained from the patient's surface motion. During binning, phase tolerances are chosen to obtain complete volumetric information since images at different couch positions are reconstructed at different respiratory phases. We describe 4DCT image formation and associated experiments that characterize the properties of 4DCT. Residual motion artifacts remain due to partial projection effects. Temporal coherence within resorted 4DCT volumes is dominated by the number of reconstructed images per slice. The more images are reconstructed, the smaller phase tolerances can be for retrospective sorting. From phantom studies a precision of about 2.5 mm for quasiregular motion and typical respiratory periods could be concluded. A protocol for 4DCT scanning was evaluated and clinically implemented at the MGH. Patient data are presented to elucidate how additional patient specific parameters can impact 4DCT imaging. PMID- 15895571 TI - Quantitation of the reconstruction quality of a four-dimensional computed tomography process for lung cancer patients. AB - We have developed a four-dimensional computed tomography (4D CT) technique for mapping breathing motion in radiotherapy treatment planning. A multislice CT scanner (1.5 mm slices) operated in cine mode was used to acquire 12 contiguous slices in each couch position for 15 consecutive scans (0.5 s rotation, 0.25 s between scans) while the patient underwent simultaneous quantitative spirometry measurements to provide a sorting metric. The spirometry-sorted scans were used to reconstruct a 4D data set. A critical factor for 4D CT is quantifying the reconstructed data set quality which we measure by correlating the metric used relative to internal-object motion. For this study, the internal air content within the lung was used as a surrogate for internal motion measurements. Thresholding and image morphological operations were applied to delineate the air containing tissues (lungs, trachea) from each CT slice. The Hounsfield values were converted to the internal air content (V). The relationship between the air content and spirometer-measured tidal volume (v) was found to be quite linear throughout the lungs and was used to estimate the overall accuracy and precision of tidal volume-sorted 4D CT. Inspection of the CT-scan air content as a function of tidal volume showed excellent correlations (typically r>0.99) throughout the lung volume. Because of the discovered linear relationship, the ratio of internal air content to tidal volume was indicative of the fraction of air change in each couch position. Theoretically, due to air density differences within the lung and in room, the sum of these ratios would equal 1.11. For 12 patients, the mean value was 1.08 +/- 0.06, indicating the high quality of spirometry-based image sorting. The residual of a first-order fit between v and V was used to estimate the process precision. For all patients, the precision was better than 8%, with a mean value of 5.1% +/- 1.9%. This quantitative analysis highlights the value of using spirometry as the metric in sorting CT scans. The 4D reconstruction provides the CT data required to measure the three-dimensional trajectory of tumor and lung tissue during free breathing. PMID- 15895572 TI - Brachytherapy needle deflection evaluation and correction. AB - In prostate brachytherapy, an 18-gauge needle is used to implant radioactive seeds. This thin needle can be deflected from the preplanned trajectory in the prostate, potentially resulting in a suboptimum dose pattern and at times requiring repeated needle insertion to achieve optimal dosimetry. In this paper, we report on the evaluation of brachytherapy needle deflection and bending in test phantoms and two approaches to overcome the problem. First we tested the relationship between needle deflection and insertion depth as well as whether needle bending occurred. Targeting accuracy was tested by inserting a brachytherapy needle to target 16 points in chicken tissue phantoms. By implanting dummy seeds into chicken tissue phantoms under 3D ultrasound guidance, the overall accuracy of seed implantation was determined. We evaluated methods to overcome brachytherapy needle deflection with three different insertion methods: constant orientation, constant rotation, and orientation reversal at half of the insertion depth. Our results showed that needle deflection is linear with needle insertion depth, and that no noticeable bending occurs with needle insertion into the tissue and agar phantoms. A 3D principal component analysis was performed to obtain the population distribution of needle tip and seed position relative to the target positions. Our results showed that with the constant orientation insertion method, the mean needle targeting error was 2.8 mm and the mean seed implantation error was 2.9 mm. Using the constant rotation and orientation reversal at half insertion depth methods, the deflection error was reduced. The mean needle targeting errors were 0.8 and 1.2 mm for the constant rotation and orientation reversal methods, respectively, and the seed implantation errors were 0.9 and 1.5 mm for constant rotation insertion and orientation reversal methods, respectively. PMID- 15895573 TI - Monte Carlo simulation of a protontherapy platform devoted to ocular melanoma. AB - Patients with ocular melanoma have been treated since June 1991 at the medical cyclotron of the Centre Antoine Lacassagne (CAL). Positions and sizes of the ocular nozzle elements were initially defined based on experimental work, taking as a pattern functional existing facilities. Nowadays Monte Carlo (MC) calculation offers a tool to refine this geometry by adjusting size and place of beam modeling devices. Moreover, the MC tool is a useful way to calculate the dose and to evaluate the impact of secondary particles in the field of radiotherapy or radiation protection. Both LINAC and cyclotron producing x rays, electrons, protons, and neutrons are available in CAL, which suggests choosing MCNPX for its particle versatility. As a first step, the existing installation was input in MCNPX to check its aptitude to reproduce experimentally measured depth-dose profile, lateral profile, output-factor (OF), and absolute dose. The geometry was defined precisely and described from the last achromatic bending magnet of our proton beam line to the position of treated eyes. Relative comparisons of percentage depth-dose and lateral profiles, performed between measured data and simulations, show an agreement of the order of 2% in dose and 0.1 mm in range accuracy. These comparisons, carried out with and without beam modifying device, yield results compatible to the required precision in ocular melanoma treatments, as long as adequate choices are made on MCNPX input decks for physics card. Absolute dose and OF issued from calculations and measurements were also compared. Results obtained for these two kinds of data, carried out in the simplified situation of an unmodulated beam, indicate that MC calculation could effectively complement measurements. These encouraging results are a large source of motivation to promote further studies, first in a new design of the ocular nozzle, and second in the analysis of the influence of beam-modifying devices attached to the final patient collimator, such as wedge or compensators, on dose values. PMID- 15895574 TI - Investigation of using a power function as a cost function in inverse planning optimization. AB - The purpose of this paper is to investigate the use of a power function as a cost function in inverse planning optimization. The cost function for each structure is implemented as an exponential power function of the deviation between the resultant dose and prescribed or constrained dose. The total cost function for all structures is a summation of the cost function of every structure. When the exponents of all terms in the cost function are set to 2, the cost function becomes a classical quadratic cost function. An independent optimization module was developed and interfaced with a research treatment planning system from the University of North Carolina for dose calculation and display of results. Three clinical cases were tested for this study with various exponents set for tumor targets and sensitive structures. Treatment plans with these exponent settings were compared, using dose volume histograms. The results of our study demonstrated that using an exponent higher than 2 in the cost function for the target achieved better dose homogeneity than using an exponent of 2. An exponent higher than 2 for serial sensitive structures can effectively reduce the maximum dose. Varying the exponent from 2 to 4 resulted in the most effective changes in dose volume histograms while the change from 4 to 8 is less drastic, indicating a situation of saturation. In conclusion, using a power function with exponent greater than 2 as a cost function can effectively achieve homogeneous dose inside the target and/or minimize maximum dose to the critical structures. PMID- 15895575 TI - Experimental validation of the Wigner distributions theory of phase-contrast imaging. AB - Recently, a new theory of phase-contrast imaging has been proposed by Wu and Liu [Med. Phys. 31, 2378-2384 (2004)]. This theory, based upon Wigner distributions, provides a much stronger foundation for the evaluation of phase-contrast imaging systems than did the prior theories based upon Fresnel-Kirchhoff diffraction theory. In this paper, we compare results of measurements made in our laboratory of phase contrast for different geometries and tube voltages to the predictions of the Wu and Liu model. In our previous publications, we have used an empirical measurement (the edge enhancement index) to parametrize the degree of phase contrast effects in an image. While the Wu and Liu model itself does not predict image contrast, it does measure the degree of phase contrast that the system can image for a given spatial frequency. We have found that our previously published experimental results relating phase-contrast effects to geometry and x-ray tube voltage are consistent with the predictions of the Wu and Liu model. PMID- 15895576 TI - Determination of maximum leaf velocity and acceleration of a dynamic multileaf collimator: implications for 4D radiotherapy. AB - The dynamic multileaf collimator (MLC) can be used for four-dimensional (4D), or tumor tracking radiotherapy. However, the leaf velocity and acceleration limitations become a crucial factor as the MLC leaves need to respond in near real time to the incoming respiration signal. The aims of this paper are to measure maximum leaf velocity, acceleration, and deceleration to obtain the mechanical response times for the MLC, and determine whether the MLC is suitable for 4D radiotherapy. MLC leaf sequence files, requiring the leaves to reach maximum acceleration and velocity during motion, were written. The leaf positions were recorded every 50 ms, from which the maximum leaf velocity, acceleration, and deceleration were derived. The dependence on the velocity and acceleration of the following variables were studied: leaf banks, inner and outer leaves, MLC-MLC variations, gravity, friction, and the stability of measurements over time. Measurement results show that the two leaf banks of a MLC behave similarly, while the inner and outer leaves have significantly different maximum leaf velocities. The MLC-MLC variations and the dependence of gravity on maximum leaf velocity are statistically significant. The average maximum leaf velocity at the isocenter plane of the MLC ranged from 3.3 to 3.9 cm/s. The acceleration and deceleration at the isocenter plane of the MLC ranged from 50 to 69 cm/s2 and 46 to 52 cm/s2, respectively. Interleaf friction had a negligible effect on the results, and the MLC parameters remained stable with time. Equations of motion were derived to determine the ability of the MLC response to fluoroscopymeasured diaphragm motion. Given the present MLC mechanical characteristics, 4D radiotherapy is feasible for up to 97% of respiratory motion. For the largest respiratory motion velocities observed, beam delivery should be temporarily stopped (beam hold). PMID- 15895577 TI - Four-dimensional radiotherapy planning for DMLC-based respiratory motion tracking. AB - Four-dimensional (4D) radiotherapy is the explicit inclusion of the temporal changes in anatomy during the imaging, planning, and delivery of radiotherapy. Temporal anatomic changes can occur for many reasons, though the focus of the current investigation is respiration motion for lung tumors. The aim of this study was to develop 4D radiotherapy treatment-planning methodology for DMLC based respiratory motion tracking. A 4D computed tomography (CT) scan consisting of a series of eight 3D CT image sets acquired at different respiratory phases was used for treatment planning. Deformable image registration was performed to map each CT set from the peak-inhale respiration phase to the CT image sets corresponding to subsequent respiration phases. Deformable registration allows the contours defined on the peak-inhale CT to be automatically transferred to the other respiratory phase CT image sets. Treatment planning was simultaneously performed on each of the eight 3D image sets via automated scripts in which the MLC-defined beam aperture conforms to the PTV (which in this case equaled the GTV due to CT scan length limitations) plus a penumbral margin at each respiratory phase. The dose distribution from each respiratory phase CT image set was mapped back to the peak-inhale CT image set for analysis. The treatment intent of 4D planning is that the radiation beam defined by the DMLC tracks the respiration induced target motion based on a feedback loop including the respiration signal to a real-time MLC controller. Deformation with respiration was observed for the lung tumor and normal tissues. This deformation was verified by examining the mapping of high contrast objects, such as the lungs and cord, between image sets. For the test case, dosimetric reductions for the cord, heart, and lungs were found for 4D planning compared with 3D planning. 4D radiotherapy planning for DMLC-based respiratory motion tracking is feasible and may offer tumor dose escalation and/or a reduction in treatment-related complications. However, 4D planning requires new planning tools, such as deformable registration and automated treatment planning on multiple CT image sets. PMID- 15895578 TI - Method for a detailed measurement of image intensity nonuniformity in magnetic resonance imaging. AB - In magnetic resonance imaging (MRI), the MR signal intensity can vary spatially and this spatial variation is usually referred to as MR intensity nonuniformity. Although the main source of intensity nonuniformity arises from B1 inhomogeneity of the coil acting as a receiver and/or transmitter, geometric distortion also alters the MR signal intensity. It is useful on some occasions to have these two different sources be separately measured and analyzed. In this paper, we present a practical method for a detailed measurement of the MR intensity nonuniformity. This method is based on the same three-dimensional geometric phantom that was recently developed for a complete measurement of the geometric distortion in MR systems. In this paper, the contribution to the intensity nonuniformity from the geometric distortion can be estimated and thus, it provides a mechanism for estimation of the intensity nonuniformity that reflects solely the spatial characteristics arising from B1. Additionally, a comprehensive scheme for characterization of the intensity nonuniformity based on the new measurement method is proposed. To demonstrate the method, the intensity nonuniformity in a 1.5 T Sonata MR system was measured and is used to illustrate the main features of the method. PMID- 15895579 TI - A new data consistency condition for fan-beam projection data. AB - The sum of all attenuation data acquired in one view of parallel-beam projections is a view angle independent constant. This fact is known as a data consistency condition on the two-dimensional Radon transforms. It plays an important role in tomographic image reconstruction and artifact correction. In this paper, a novel fan-beam data consistency condition (FDCC) is derived and presented. Using the FDCC, individual projection data in one view of fan-beam projections can be estimated from filtering all other projection data measured from different view angles. Numerical simulations are performed to validate the new FDCC in correcting ring artifacts caused by malfunctioning detector cells. PMID- 15895580 TI - Accurate technique for complete geometric calibration of cone-beam computed tomography systems. AB - Cone-beam computed tomography systems have been developed to provide in situ imaging for the purpose of guiding radiation therapy. Clinical systems have been constructed using this approach, a clinical linear accelerator (Elekta Synergy RP) and an iso-centric C-arm. Geometric calibration involves the estimation of a set of parameters that describes the geometry of such systems, and is essential for accurate image reconstruction. We have developed a general analytic algorithm and corresponding calibration phantom for estimating these geometric parameters in cone-beam computed tomography (CT) systems. The performance of the calibration algorithm is evaluated and its application is discussed. The algorithm makes use of a calibration phantom to estimate the geometric parameters of the system. The phantom consists of 24 steel ball bearings (BBs) in a known geometry. Twelve BBs are spaced evenly at 30 deg in two plane-parallel circles separated by a given distance along the tube axis. The detector (e.g., a flat panel detector) is assumed to have no spatial distortion. The method estimates geometric parameters including the position of the x-ray source, position, and rotation of the detector, and gantry angle, and can describe complex source-detector trajectories. The accuracy and sensitivity of the calibration algorithm was analyzed. The calibration algorithm estimates geometric parameters in a high level of accuracy such that the quality of CT reconstruction is not degraded by the error of estimation. Sensitivity analysis shows uncertainty of 0.01 degrees (around beam direction) to 0.3 degrees (normal to the beam direction) in rotation, and 0.2 mm (orthogonal to the beam direction) to 4.9 mm (beam direction) in position for the medical linear accelerator geometry. Experimental measurements using a laboratory bench Cone-beam CT system of known geometry demonstrate the sensitivity of the method in detecting small changes in the imaging geometry with an uncertainty of 0.1 mm in transverse and vertical (perpendicular to the beam direction) and 1.0 mm in the longitudinal (beam axis) directions. The calibration algorithm was compared to a previously reported method, which uses one ball bearing at the isocenter of the system, to investigate the impact of more precise calibration on the image quality of cone beam CT reconstruction. A thin steel wire located inside the calibration phantom was imaged on the conebeam CT lab bench with and without perturbations in source and detector position during the scan. The described calibration method improved the quality of the image and the geometric accuracy of the object reconstructed, improving the full width at half maximum of the wire by 27.5% and increasing contrast of the wire by 52.8%. The proposed method is not limited to the geometric calibration of cone-beam CT systems but can be used for many other systems, which consist of one or more point sources and area detectors such as calibration of megavoltage (MV) treatment system (focal spot movement during the beam delivery, MV source trajectory versus gantry angle, the axis of collimator rotation, and couch motion), cross calibration between Kilovolt imaging and MV treatment system, and cross calibration between multiple imaging systems. Using the complete information of the system geometry, it was demonstrated that high image quality in CT reconstructions is possible even in systems with large geometric nonidealities. PMID- 15895581 TI - Respiratory motion estimation from slowly rotating x-ray projections: theory and simulation. AB - Understanding the movement of tumors caused by respiratory motion is very important for conformal radiatherapy. However, respiratory motion is very difficult to study by conventional x-ray CT imaging since object motion causes inconsistent projection views, leading to artifacts in reconstructed images. We propose to estimate the parameters of a nonrigid, free breathing motion model from a set of projection views of the thorax that are acquired using a slowly rotating cone-beam CT scanner. This approach involves deforming a motion-free reference thorax volume according to the estimated parameters and comparing its projections to the corresponding measured projection views. The parameters are optimized by minimizing a regularized squared error cost function. Simulation results with a fan-beam geometry show good agreement between the estimated motion and the true motion, which supports the potential of this approach for estimating four-dimensional (three-dimensional spatial + temporal) respiratory motion. PMID- 15895582 TI - Three-dimensional fluorescence lifetime tomography. AB - Near-infrared fluorescence tomography using molecularly targeted lifetime sensitive, fluorescent contrast agents have applications for early-stage cancer diagnostics. Yet, although the measurement of fluorescent lifetime imaging microscopy (FLIM) is extensively used in microscopy and spectroscopy applications, demonstration of fluorescence lifetime tomography for medical imaging is limited to two-dimensional studies. Herein, the feasibility of three dimensional fluorescence-lifetime tomography on clinically relevant phantom volumes is established, using (i) a gainmodulated intensified charge coupled device (CCD) and modulated laser diode imaging system, (ii) two fluorescent contrast agents, e.g., Indocyanine green and 3-3'-Diethylthiatricarbocyanine iodide differing in their fluorescence lifetime by 0.62 ns, and (iii) a two stage approximate extended Kalman filter reconstruction algorithm. Fluorescence measurements of phase and amplitude were acquired on the phantom surface under different target to background fluorescence absorption (70:1, 100:1) and fluorescence lifetime (1:1, 2.1:1) contrasts at target depths of 1.4-2 cm. The Bayesian tomography algorithm was employed to obtain three-dimensional images of lifetime and absorption owing to the fluorophores. PMID- 15895583 TI - ROC study of the effect of stereoscopic imaging on assessment of breast lesions. AB - An observer performance study was conducted to evaluate the usefulness of assessing breast lesion characteristics with stereomammography. Stereoscopic image pairs of 158 breast biopsy tissue specimens were acquired with a GE Senographe 2000D full field digital mammography system using a 1.8x magnification geometry. A phantom-shift method equivalent to a stereo shift angle of +/- 3 degrees relative to a central axis perpendicular to the detector was used. For each specimen, two pairs of stereo images were taken at approximately orthogonal orientations. The specimens contained either a mass, microcalcifications, both, or normal tissue. Based on pathological analysis, 39.9% of the specimens were found to contain malignancy. The digital specimen radiographs were displayed on a high resolution MegaScan CRT monitor driven by a DOME stereo display board using in-house developed software. Five MQSA radiologists participated as observers. Each observer read the 316 specimen stereo image pairs in a randomized order. For each case, the observer first read the monoscopic image and entered his/her confidence ratings on the presence of microcalcifications and/or masses, margin status, BI-RADS assessment, and the likelihood of malignancy. The corresponding stereoscopic images were then displayed on the same monitor and were viewed through stereoscopic LCD glasses. The observer was free to change the ratings in every category after stereoscopic reading. The ratings of the observers were analyzed by ROC methodology. For the 5 MQSA radiologists, the average Az value for estimation of the likelihood of malignancy of the lesions improved from 0.70 for monoscopic reading to 0.72 (p=0.04) after stereoscopic reading, and the average Az value for the presence of microcalcifications improved from 0.95 to 0.96 (p=0.02). The Az value for the presence of masses improved from 0.80 to 0.82 after stereoscopic reading, but the difference fell short of statistical significance (p=0.08). The visual assessment of margin clearance was found to have very low correlation with microscopic analysis with or without stereoscopic reading. This study demonstrates the potential of using stereomammography to improve the detection and characterization of mammographic lesions. PMID- 15895584 TI - The calibration of experimental self-developing Gafchromic HXR film for the measurement of radiation dose in computed tomography. AB - A prototype, self-developing Gafchromic HXR film has sensitivity an order of magnitude larger than that of the commercially available Gafchromic XR film used in interventional radiological applications. The higher sensitivity of the HXR film allows the possibility of acquisition of high-resolution calibrated dose profiles within the diagnostic range of exposure levels, below 10 R (87.7 mGy). We employed a commercially available, optical flatbed scanner for digitization of the film and image analysis software to determine the response of the HXR films to ionizing radiation. Spatial uniformity and temporal repeatability of the flatbed scanner were determined and used in optimization of the digitization protocol. The HXR film postexposure density growth and sensitivity to ambient light were determined using multiple scans of two simultaneously exposed sheets, one stored in light-tight conditions and the other continuously illuminated with white light. A calibrated step wedge of the HXR film was obtained by simultaneous irradiation of a portion of a film strip and a calibrated ionization chamber using a radiographic x-ray tube with beam characteristics matched to a typical CT scanner (8 mm Al HVL, 120 kVp). Repeated digitization of the calibration film was used to determine the precision of the film response measurements. The precision, as measured by the standard deviation of multiple measurements, was better than 1% over the full dynamic range of film response. This precision was measured using exposures ranging from 0.5 to 12 R (4.4 to 105.3 mGy). This exposure range is highly relevant to x-ray computed tomography. Preliminary radiation dose profiles demonstrate the utility of this technique. PMID- 15895585 TI - A least-squares error minimization approach in the determination of ferric ion diffusion coefficient of Fricke-infused dosimeter gels. AB - A least-squares error minimization approach was adopted to assess ferric ion diffusion coefficient of Fricke-agarose gels. Ferric ion diffusion process was modeled as a Gaussian-shaped degradation kernel operating on an initial concentration distribution. Diffusion coefficient was iteratively determined by minimizing the error function defined as the difference between the theoretically calculated and the experimentally measured dose distributions. A rapid MR image based differential gel dosimetry technique that time resolves the evolution of the ferric ion diffusion process minimizes smearing of the dose distribution. Our results showed that for a Fricke-agarose gel contained 1 mM ammonium ferrous sulfate, 1% agarose, 1 mM sodium chloride, and 50 mM sulfuric acid, its ferric ion diffusion coefficient is (1.59 +/- 0.28) x 10(-2) cm2 h(-1) at room temperature. This value falls within the 1.00-2.00 x 10(-2) cm2 h(-1) range previously reported under varying gelling ingredients and concentrations. This method allows a quick, nondestructive evaluation of the ferric ion diffusion coefficient that can be used in conjunction with the in situ gel dosimetry experiment to provide a practical diffusion characterization of the dosimeter gel. PMID- 15895586 TI - Reduction of eye lens radiation dose by orbital bismuth shielding in pediatric patients undergoing CT of the head: a Monte Carlo study. AB - Our aim in the study was to assess the eye lens dose reduction resulting from the use of radioprotective bismuth garments to shield the eyes of pediatric patients undergoing head CT. The Monte Carlo N-particle transport code and mathematical humanoid phantoms representing the average individual at different ages were used to determine eye lens dose reduction accomplished with bismuth shielding of the eye in the following simulated CT scans: (a) scanning of the orbits, (b) scanning of the whole head, and (c) 20 degrees angled scanning of the brain excluding the orbits. The effect of bismuth shielding on the eye lens dose was also investigated using an anthropomorphic phantom and thermoluminescence dosimetry (TLD). Eye lens dose reduction achieved by bismuth shielding was measured in 16 patients undergoing multiphase CT scanning of the head. The patient's scans were divided in the following: CT examinations where the eye globes were entirely included (n=5), partly included (n=6) and excluded (n=5) from the scanned region. The eye lens dose reduction depended mainly on the scan boundaries set by an operator. The average eye lens dose reduction determined by Monte Carlo simulation was 38.2%, 33.0% and <1% for CT scans of the orbits, whole head, and brain with an angled gantry, respectively. The difference between the Monte Carlo derived eye lens dose reduction factor values and corresponding values determined directly by using the anthropomorphic phantom head was found less than 5%. The mean eye lens dose reduction achieved by bismuth shielding in pediatric patients were 34%, 20% and <2% when eye globes were entirely included, partly included and excluded from the scanned region, respectively. A significant reduction in eye lens dose may be achieved by using superficial orbital bismuth shielding during pediatric head CT scans. However, bismuth garments should not be used in children when the eyes are excluded from the primarily exposed region. PMID- 15895587 TI - A comparison of two data analyses from two observer performance studies using Jackknife ROC and JAFROC. AB - The authors compared two methodological approaches, Jackknife ROC and JAFROC, in analyzing data ascertained during FROC (free-response receiver operating characteristics) type studies. Observer rating data obtained from two observer performance studies were analyzed. During the first study, seven radiologists interpreted 120 mammography examinations depicting 57 masses under five different conditions with and without the results of computer-aided detection (CAD). In the second study, eight radiologists interpreted 110 examinations depicting 51 masses under six different display conditions with and without CAD results. Readers rated the detection task in a FROC type response. Jackknife ROC (using the software of LABMRMC with the highest rating per case) and JAFROC were used to compute differences, if any, in summary performance levels among all reading modes in each study as well as for all paired data sets. The results of the different analytical approaches are compared. The overall results for all modes were significantly different for the first study (p < 0.05) and not significant (p > 0.05) for the second study using either analytical approach. In the first study, the performance levels represented by three paired data sets were significantly different (p < 0.05) when computed using LABMRMC and four pairs were significantly different (p < 0.05) using JAFROC. In eight of ten pairs, JAFROC produced lower p values than LABMRMC. In the second study, LABMRMC showed no significant differences for any paired data sets and JAFROC showed a significant difference for one pair. In 15 of 16 pairs, p values computed by JAFROC were lower than those computed by LABMRMC. PMID- 15895588 TI - Density resolution of proton computed tomography. AB - Conformal proton radiation therapy requires accurate prediction of the Bragg peak position. Protons may be more suitable than conventional x-rays for this task since the relative electron density distribution can be measured directly with proton computed tomography (CT). However, proton CT has its own limitations, which need to be carefully studied before this technique can be introduced into routine clinical practice. In this work, we have used analytical relationships as well as the Monte Carlo simulation tool GEANT4 to study the principal resolution limits of proton CT. The noise level observed in proton CT images of a cylindrical water phantom with embedded tissue-equivalent density inhomogeneities, which were generated based on GEANT4 simulations, compared well with predictions based on Tschalar's theory of energy loss straggling. The relationship between phantom thickness, initial energy, and the relative electron density resolution was systematically investigated to estimate the proton dose needed to obtain a given density resolution. We show that a reasonable density resolution can be achieved with a relatively small dose, which is comparable to or even lower than that of x-ray CT. PMID- 15895589 TI - Radiological properties of normoxic polymer gel dosimeters. AB - The radiological properties of the normoxic polymer gel dosimeters MAGIC, MAGAS, and MAGAT [methacrylic and ascorbic acid in gelatin initiated by copper; methacrylic acid gelatine gel with ascorbic acid; and methacrylic acid gelatine and tetrakis (hydroxymethyl) phosphonium chloride, respectively] have been investigated. The radiological water equivalence was determined by comparing the polymer gel macroscopic photon and electron interaction cross sections over the energy range from 10 keV to 20 MeV and by Monte Carlo modeling of depth doses. Normoxic polymer gel dosimeters have a high gelatine and monomer concentration and therefore mass density (kg m(-3)) up to 3.8% higher than water. This results in differences between the cross-section ratios of the normoxic polymer gels and water of up to 3% for the attenuation, energy absorption, and collision stopping power coefficient ratios through the Compton dominant energy range. The mass cross-section ratios were within 2% of water except for the mass attenuation and energy absorption coefficients ratios, which showed differences with water of up to 6% for energies less than 100 keV. Monte Carlo modeling was undertaken for the polymer gel dosimeters to model the electron and photon transport resulting from a 6 MV photon beam. The absolute percentage differences between gel and water were within 1% and the relative percentage differences were within 3.5%. The results show that the MAGAT gel formulation is the most radiological water equivalent of the normoxic polymer gel dosimeters investigated due to its lower mass density measurement compared with MAGAS and MAGIC gels. PMID- 15895590 TI - Low dose fraction behavior of high sensitivity radiochromic film. AB - A high sensitivity (HS) model of radiochromic film is receiving increasing use. The film's linear sensitometric response in the range of 0.5-40 Gy would make this film an ideal candidate for complex dosimetry applications that require tissue equivalence. This study investigates the potential use for clinical dosimetry of typical radiotherapy fractions at relatively low doses (0.5-5 Gy). The experiment involved exposing 25 pre-exposed pieces of HS film to five equal fractions of doses from 0.5 to 5 Gy 24 hours apart. The cumulative dose for each film was carefully monitored and optical density measurements were used as the sole determination of film response to dose. The average behavior of the various fractionation schemes was roughly consistent with previous observations of the MD 55 radiochromic film with about twice the overall sensitivity as expected. However, at low doses and low dose increments, unexpected variations beyond a well-documented low dose nonlinearity were observed. These unexpected variations may indicate complex polymer kinetics at low doses. This type of film would require extra care beyond that described in TG-55 for accurate use at low doses or low dose fraction schemes. PMID- 15895591 TI - Enlarged longitudinal dose profiles in cone-beam CT and the need for modified dosimetry. AB - In order to examine phantom length necessary to assess radiation dose delivered to patients in cone-beam CT with an enlarged beamwidth, we measured dose profiles in cylindrical phantoms of sufficient length using a prototype 256-slice CT scanner developed at our institute. Dose profiles parallel to the rotation axis were measured at the central and peripheral positions in PMMA (polymethylmethacrylate) phantoms of 160 or 320 mm diameter and 900 mm length. For practical application, we joined unit cylinders (150 mm long) together to provide phantoms of 900 mm length. Dose profiles were measured with a pin photodiode sensor having a sensitive region of approximately 2.8 x 2.8 mm2 and 2.7 mm thickness. Beamwidths of the scanner were varied from 20 to 138 mm. Dose profile integrals (DPI) were calculated using the measured dose profiles for various beamwidths and integration ranges. For the body phantom (320-mm-diam phantom), 76% of the DPI was represented for a 20 mm beamwidth and 60% was represented for a 138 mm beamwidth if dose profiles were integrated over a 100 mm range, while more than 90% of the DPI was represented for beamwidths between 20 and 138 mm if integration was carried out over a 300 mm range. The phantom length and integration range for dosimetry of cone-beam CT needed to be more than 300 mm to represent more than 90% of the DPI for the body phantom with the beamwidth of more than 20 mm. Although we reached this conclusion using the prototype 256 slice CT-scanner, it may be applied to other multislice CT-scanners as well. PMID- 15895592 TI - Distributions of beta+ decayed nuclei generated in the CH2 and H2O targets by the target nuclear fragment reaction using therapeutic MONO and SOBP proton beam. AB - In proton radiotherapy, the irradiation dose can be concentrated on a tumor. To use this radiotherapy efficiently in the clinical field, it is necessary to evaluate the proton-irradiated area and condition. The proton-irradiated area can be confirmed by coincidence detection of pair annihilation gamma rays from beta+ decayed nuclei generated by target nuclear fragment reaction of irradiated proton nuclei and nuclei in the irradiation target. In this study, we performed experiments of proton irradiation to a polyethylene (PE:CH2) target containing 12C nuclei, which is a major component of the human body, and a gelatinous water (H2O) target containing 16O nuclei at different proton irradiation energy levels under different beam conditions of mono-energetic Bragg peak and spread-out Bragg peak. The distribution of the activity in the target after proton irradiation was measured by a positron emission tomography (PET) apparatus, and compared with the calculated distribution. The temporal dependence of the activity distribution during the period between the completion of proton irradiation and the start of measurement by the PET apparatus was examined. The activity by clinical proton irradiation was 3 kB/cc in the PE target and 13 kB/cc in the water target, indicating that the intensity was sufficient for the evaluation of the distribution. The range of the activity distribution against the physical range was short (several millimeter water equivalent length), indicating the presence of target dependence. The range difference in the water target was slightly large with time dependence until the start of measurement. The difference of the lateral widths with full width half at maximum in the distributions of the measured irradiated dose and activity was within 1 mm. PMID- 15895593 TI - Improved accuracy and consistency in T1 measurement of flowing blood by using inversion recovery GE-EPI. AB - Problems associated with techniques currently used to measure the T1 of flowing blood are evaluated and a method to improve the consistency and repeatability of measurements is presented. Similar to some currently used techniques, the pulse sequence employs a nonselective adiabatic inversion pulse followed by a series of ECG-gated gradient echo EPI (echo planar imaging) images to obtain images where the blood (fluid) signal exhibits a T1-dependent inversion recovery signal from which the spin lattice relaxation constant (T1) of the flowing fluid can be measured. The new method combines curve fitting with a measure of the curve null point to acquire more accurate and consistent T1 values. Simulation and experimental results show that this combined fitting-nulling method is more stable and consistent in measuring the T1 of flowing fluid. The feasibility of temperature measurement of a flowing fluid based on the temperature dependence of the T1 of water protons is shown in this paper. ECG gating is used to reduce the effects of cyclic intensity changes for measurement of T1 in pulsatile flowing blood. PMID- 15895594 TI - Near-real-time radiotherapy dosimetry using optically stimulated luminescence of Al2O3:C: mathematical models and preliminary results. AB - In this paper we report investigations aimed toward applying optically stimulated luminescence (OSL) of Al2O3:C for near-real-time medical dosimetry, especially in radiotherapy. The classical mathematical model normally used for the description of OSL phenomena was expanded to predict the behavior of the luminescence signal in the case when the OSL sample is simultaneously irradiated and optically stimulated. The predictions obtained were used to develop different measurement approaches and correction algorithms for the luminescence signals, thus enabling dose estimation from OSL during rather then after the irradiation procedure. Radiation probes with diameters of less than 1 mm, suitable for the envisioned in vivo measurements were constructed by attaching small Al2O3:C crystals to optical fiber cables. The OSL fiber probes and a purpose-built, portable OSL stimulation and readout system were used to measure doses at speeds up to 1 data point every 3s, under irradiation at dose rates of the same order of magnitude as those found in conventional radiotherapy techniques. The corrected OSL signal was found to be proportional to the absorbed dose, and accurately followed sudden transitions in the irradiation dose rate. PMID- 15895595 TI - The effect of ambient pressure on well chamber response: Monte Carlo calculated results for the HDR 1000 plus. AB - The determination of the air kerma strength of a brachytherapy seed is necessary for effective treatment planning. Well ionization chambers are used on site at therapy clinics to determine the air kerma strength of seeds. In this work, the response of the Standard Imaging HDR 1000 Plus well chamber to ambient pressure is examined using Monte Carlo calculations. The experimental work examining the response of this chamber as well as other chambers is presented in a companion paper. The Monte Carlo results show that for low-energy photon sources, the application of the standard temperature pressure PTP correction factor produces an over-response at the reduced air densities/pressures corresponding to high elevations. With photon sources of 20 to 40 keV, the normalized PTP corrected chamber response is as much as 10% to 20% over unity for air densities/pressures corresponding to an elevation of 3048 m (10000 ft) above sea level. At air densities corresponding to an elevation of 1524 m (5000 ft), the normalized PTP corrected chamber response is 5% to 10% over unity for these photon sources. With higher-energy photon sources (>100 keV), the normalized PTP corrected chamber response is near unity. For low-energy beta sources of 0.25 to 0.50 MeV, the normalized PTP-corrected chamber response is as much as 4% to 12% over unity for air densities/pressures corresponding to an elevation of 3048 m (10000 ft) above sea level. Higher-energy beta sources (>0.75 MeV) have a normalized PTP corrected chamber response near unity. Comparing calculated and measured chamber responses for common 103Pd- and 125I-based brachytherapy seeds show agreement to within 2.7% and 1.9%, respectively. Comparing MCNP calculated chamber responses with EGSnrc calculated chamber responses show agreement to within 3.1% at photon energies of 20 to 40 keV. We conclude that Monte Carlo transport calculations accurately model the response of this well chamber. Further, applying the standard PTP correction factor for this well chamber is insufficient in accounting for the change in chamber response with air pressure for low-energy (<100 keV) photon and low-energy (<0.75 MeV)beta sources. PMID- 15895596 TI - Comprehensive Monte Carlo calculation of the point spread function for a commercial a-Si EPID. AB - Images produced by commercial amorphous silicon electronic portal imaging devices (a-Si EPIDs) are subject to multiple blurring processes. Implementation of these devices for fluence measurement requires that the blur be removed from the images. A standard deconvolution operation can be performed to accomplish this assuming the blur kernel is spatially invariant and accurately known. This study determines a comprehensive blur kernel for the Varian aS500 EPID. Monte Carlo techniques are used to derive a dose kernel and an optical kernel, which are then combined to yield an overall blur kernel for both 6 and 15 MV photon beams. Experimental measurement of the line spread function (LSF) is used to verify kernel shape. Kernel performance is gauged by comparing EPID image profiles with in-air dose profiles measured using a diamond detector (approximating fluence) both before and after the EPID images have been deconvolved. Quantitative comparisons are performed using the chi metric, an extension of the well-known y metric, using acceptance criteria of 0.0784 cm (1 pixel width) distance-to agreement (deltad) and 2% of the relative central axis fluence (deltaD). Without incorporating any free parameters, acceptance was increased from 49.0% of pixels in a cross-plane profile for a 6 MV 10 x 10 cm2 open field to 92.0%. For a 10 x 10 cm2 physically wedged field, acceptance increased from 40.3% to 73.9%. The effect of the optical kernel was found to be negligible for these chi acceptance parameters, however for (deltaD= 1%, deltad = 0.0784 cm) we observed an improvement from 66.1% (without) to 78.6% (with) of chi scores <1 (from 20.6% before deconvolution). It is demonstrated that an empirical kernel having a triple exponential form or a semiempirical kernel based on a simplified model of the detector stack can match the performance of the comprehensive kernel. PMID- 15895597 TI - Diffuse optical tomography of breast cancer during neoadjuvant chemotherapy: a case study with comparison to MRI. AB - We employ diffuse optical tomography (DOT) to track treatment progress in a female subject presenting with locally advanced invasive carcinoma of the breast during neoadjuvant chemotherapy. Three-dimensional images of total hemoglobin concentration and scattering identified the tumor. Our measurements reveal tumor shrinkage during the course of chemotherapy, in reasonable agreement with magnetic resonance images of the same subject. A decrease in total hemoglobin concentration contrast between tumor and normal tissue was also observed over time. The results demonstrate the potential of DOT for measuring physiological parameters of breast lesions during chemotherapy. PMID- 15895598 TI - Suitability of radiochromic medium for real-time optical measurements of ionizing radiation dose. AB - A system, consisting of a novel optical fiber-based readout configuration and model-based method, has been developed to test suitability of a certain radiochromic medium for real-time measurements of ionizing radiation dose. Using this system with the radiochromic film allowed dose measurements to be performed during, and immediately after, exposure. The rates of change in OD before, during, and after exposure differ, and the change in OD during exposure was found to be proportional to applied dose in the tested range of 0-4 Gy. Estimating applied dose within an average error of less than 5% did not require a waiting time of 24-48 h as generally recommended with this radiochromic film. The errors can be further reduced by performing a calibration for each individual dosimeter setup instead of relying on batch calibration. Measurements of change in OD were found to be independent of dose-rate in the 95-570 cGy/min range for applied dose of 1 Gy or less. Some error was introduced due to dose-rate variation for doses of 2 Gy and above. The major limiting factor in utilizing this radiation sensitive medium for real-time in vivo dosimetry is the strong dependence on temperature in the clinically relevant range of 20-38 degrees C. PMID- 15895599 TI - A stochastic convolution/superposition method with isocenter sampling to evaluate intrafraction motion effects in IMRT. AB - Current methods to calculate dose distributions with organ motion can be broadly classified as "dose convolution" and "fluence convolution" methods. In the former, a static dose distribution is convolved with the probability distribution function (PDF) that characterizes the motion. However, artifacts are produced near the surface and around inhomogeneities because the method assumes shift invariance. Fluence convolution avoids these artifacts by convolving the PDF with the incident fluence instead of the patient dose. In this paper we present an alternative method that improves the accuracy, generality as well as the speed of dose calculation with organ motion. The algorithm starts by sampling an isocenter point from a parametrically defined space curve corresponding to the patient specific motion trajectory. Then a photon is sampled in the linac head and propagated through the three-dimensional (3-D) collimator structure corresponding to a particular MLC segment chosen randomly from the planned IMRT leaf sequence. The photon is then made to interact at a point in the CT-based simulation phantom. Randomly sampled monoenergetic kernel rays issued from this point are then made to deposit energy in the voxels. Our method explicitly accounts for MLC specific effects (spectral hardening, tongue-and-groove, head scatter) as well as changes in SSD with isocentric displacement, assuming that the body moves rigidly with the isocenter. Since the positions are randomly sampled from a continuum, there is no motion discretization, and the computation takes no more time than a static calculation. To validate our method, we obtained ten separate film measurements of an IMRT plan delivered on a phantom moving sinusoidally, with each fraction starting with a random phase. For 2 cm motion amplitude, we found that a ten-fraction average of the film measurements gave an agreement with the calculated infinite fraction average to within 2 mm in the isodose curves. The results also corroborate the existing notion that the interfraction dose variability due to the interplay between the MLC motion and breathing motion averages out over typical multifraction treatments. Simulation with motion waveforms more representative of real breathing indicate that the motion can produce penumbral spreading asymmetric about the static dose distributions. Such calculations can help a clinician decide to use, for example, a larger margin in the superior direction than in the inferior direction. In the paper we demonstrate that a 15 min run on a single CPU can readily illustrate the effect of a patient-specific breathing waveform, and can guide the physician in making informed decisions about margin expansion and dose escalation. PMID- 15895600 TI - Photon-beam subsource sensitivity to the initial electron-beam parameters. AB - One limitation to the widespread implementation of Monte Carlo (MC) patient dose calculation algorithms for radiotherapy is the lack of a general and accurate source model of the accelerator radiation source. Our aim in this work is to investigate the sensitivity of the photon-beam subsource distributions in a MC source model (with target, primary collimator, and flattening filter photon subsources and an electron subsource) for 6- and 18-MV photon beams when the energy and radial distributions of initial electrons striking a linac target change. For this purpose, phase-space data (PSD) was calculated for various mean electron energies striking the target, various normally distributed electron energy spread, and various normally distributed electron radial intensity distributions. All PSD was analyzed in terms of energy, fluence, and energy fluence distributions, which were compared between the different parameter sets. The energy spread was found to have a negligible influence on the subsource distributions. The mean energy and radial intensity significantly changed the target subsource distribution shapes and intensities. For the primary collimator and flattening filter subsources, the distribution shapes of the fluence and energy fluence changed little for different mean electron energies striking the target, however, their relative intensity compared with the target subsource change, which can be accounted for by a scaling factor. This study indicates that adjustments to MC source models can likely be limited to adjusting the target subsource in conjunction with scaling the relative intensity and energy spectrum of the primary collimator, flattening filter, and electron subsources when the energy and radial distributions of the initial electron-beam change. PMID- 15895601 TI - Respiratory correlated cone beam CT. AB - A cone beam computed tomography (CBCT) scanner integrated with a linear accelerator is a powerful tool for image guided radiotherapy. Respiratory motion, however, induces artifacts in CBCT, while the respiratory correlated procedures, developed to reduce motion artifacts in axial and helical CT are not suitable for such CBCT scanners. We have developed an alternative respiratory correlated procedure for CBCT and evaluated its performance. This respiratory correlated CBCT procedure consists of retrospective sorting in projection space, yielding subsets of projections that each corresponds to a certain breathing phase. Subsequently, these subsets are reconstructed into a four-dimensional (4D) CBCT dataset. The breathing signal, required for respiratory correlation, was directly extracted from the 2D projection data, removing the need for an additional respiratory monitor system. Due to the reduced number of projections per phase, the contrast-to-noise ratio in a 4D scan reduced by a factor 2.6-3.7 compared to a 3D scan based on all projections. Projection data of a spherical phantom moving with a 3 and 5 s period with and without simulated breathing irregularities were acquired and reconstructed into 3D and 4D CBCT datasets. The positional deviations of the phantoms center of gravity between 4D CBCT and fluoroscopy were small: 0.13 +/- 0.09 mm for the regular motion and 0.39 +/- 0.24 mm for the irregular motion. Motion artifacts, clearly present in the 3D CBCT datasets, were substantially reduced in the 4D datasets, even in the presence of breathing irregularities, such that the shape of the moving structures could be identified more accurately. Moreover, the 4D CBCT dataset provided information on the 3D trajectory of the moving structures, absent in the 3D data. Considerable breathing irregularities, however, substantially reduces the image quality. Data presented for three different lung cancer patients were in line with the results obtained from the phantom study. In conclusion, we have successfully implemented a respiratory correlated CBCT procedure yielding a 4D dataset. With respiratory correlated CBCT on a linear accelerator, the mean position, trajectory, and shape of a moving tumor can be verified just prior to treatment. Such verification reduces respiration induced geometrical uncertainties, enabling safe delivery of 4D radiotherapy such as gated radiotherapy with small margins. PMID- 15895602 TI - Measuring blood flow velocities based on three image processing techniques. AB - The classification of microcirculation can be based on the size of the capillary and the velocity of the blood flow. For each type of microcirculation, the ad hoc method is required to measure the blood flow velocity. In this paper, the correlation method is used to measure blood flow velocity in the small-size capillary. The large-size capillaries are classified into three types based on the blood velocities, say low-speed, high-speed, and unstable one. The template matching algorithm, the Fast Fourier Transform algorithm and a high-speed video camera measurement are used to measure the blood velocities corresponding to those three types of large-size capillary. It was shown by several experiments that fairly good results for different types of capillary may be obtained by using the proposed three image processing algorithms. PMID- 15895603 TI - Physical characteristics of a low-dose gas microstrip detector for orthopedic x ray imaging. AB - A new scanning slit gas detector dedicated to orthopedic x-ray imaging is presented and evaluated in terms of its fundamental imaging characteristics. The system is based on the micromesh gaseous structure detector and achieves primary signal amplification through electronic avalanche in the gas. This feature, together with high quantum detection efficiency and fan-beam geometry, allows for imaging at low radiation levels. The system is composed of 1764 channels spanning a width of 44.8 cm and is capable of imaging an entire patient at speeds of up to 15 cm/s. The resolution was found to be anisotropic and significantly affected by the beam quality in the horizontal direction, but otherwise sufficient for orthopedic studies. As a consequence of line-by-line acquisition, the images contain some ripple components due to mechanical vibrations combined with variations in the x-ray tube output power. The reported detective quantum efficiency (DQE) values are relatively low (0.14 to 0.20 at 0.5 mm(-1)) as a consequence of a suboptimal collimation geometry. The DQE values were found to be unaffected by the exposure down to 7 microGy, suggesting that the system is quantum limited even for low radiation levels. A system composed of two orthogonal detectors is already in use and can produce dual-view full body scans at low doses. This device could contribute to reduce the risk of radiation induced cancer in sensitive clientele undergoing intensive x-ray procedures, like young scoliotic women. PMID- 15895604 TI - Assessment of display performance for medical imaging systems: executive summary of AAPM TG18 report. AB - Digital imaging provides an effective means to electronically acquire, archive, distribute, and view medical images. Medical imaging display stations are an integral part of these operations. Therefore, it is vitally important to assure that electronic display devices do not compromise image quality and ultimately patient care. The AAPM Task Group 18 (TG18) recently published guidelines and acceptance criteria for acceptance testing and quality control of medical display devices. This paper is an executive summary of the TG18 report. TG18 guidelines include visual, quantitative, and advanced testing methodologies for primary and secondary class display devices. The characteristics, tested in conjunction with specially designed test patterns (i.e., TG18 patterns), include reflection, geometric distortion, luminance, the spatial and angular dependencies of luminance, resolution, noise, glare, chromaticity, and display artifacts. Geometric distortions are evaluated by linear measurements of the TG18-QC test pattern, which should render distortion coefficients less than 2%/5% for primary/secondary displays, respectively. Reflection measurements include specular and diffuse reflection coefficients from which the maximum allowable ambient lighting is determined such that contrast degradation due to display reflection remains below a 20% limit and the level of ambient luminance (Lamb) does not unduly compromise luminance ratio (LR) and contrast at low luminance levels. Luminance evaluation relies on visual assessment of low contrast features in the TG18-CT and TG18-MP test patterns, or quantitative measurements at 18 distinct luminance levels of the TG18-LN test patterns. The major acceptable criteria for primary/ secondary displays are maximum luminance of greater than 170/100 cd/m2, LR of greater than 250/100, and contrast conformance to that of the grayscale standard display function (GSDF) of better than 10%/20%, respectively. The angular response is tested to ascertain the viewing cone within which contrast conformance to the GSDF is better than 30%/60% and LR is greater than 175/70 for primary/secondary displays, or alternatively, within which the on axis contrast thresholds of the TG18-CT test pattern remain discernible. The evaluation of luminance spatial uniformity at two distinct luminance levels across the display faceplate using TG18-UNL test patterns should yield nonuniformity coefficients smaller than 30%. The resolution evaluation includes the visual scoring of the CX test target in the TG18-QC or TG18-CX test patterns, which should yield scores greater than 4/6 for primary/secondary displays. Noise evaluation includes visual evaluation of the contrast threshold in the TG18-AFC test pattern, which should yield a minimum of 3/2 targets visible for primary/secondary displays. The guidelines also include methodologies for more quantitative resolution and noise measurements based on MTF and NPS analyses. The display glare test, based on the visibility of the low-contrast targets of the TG18-GV test pattern or the measurement of the glare ratio (GR), is expected to yield scores greater than 3/1 and GRs greater than 400/150 for primary/secondary displays. Chromaticity, measured across a display faceplate or between two display devices, is expected to render a u',v' color separation of less than 0.01 for primary displays. The report offers further descriptions of prior standardization efforts, current display technologies, testing prerequisites, streamlined procedures and timelines, and TG18 test patterns. PMID- 15895605 TI - Polycyclic aromatic hydrocarbons in smoke from different woods and their transfer during traditional smoking into chorizo sausages with collagen and tripe casings. AB - Polycyclic aromatic hydrocarbons (PAHs), at least those with high molecular mass, are classified as probable human carcinogens by the US Environmental Protection Agency. The importance of the material used to generate smoke is indicated by the attention that the European Union is paying to list the wood that can be used to produce smoking flavour agents. The paper examines the dependence on the nature of the wood used for smoking on the formation of eight PAHs (selected as markers of PAHs) and transfer of these PAHs into traditional Spanish smoked chorizo sausages with collagen and tripe casings. The results showed that the kind of combustion performed on different materials seriously affected the PAH levels in the smoke generated. Ignition and firing of the material with a flame compared with heating produced more PAHs in the smoke. The levels of PAHs found in the meat and in the collagen and tripe casings of the smoked chorizo samples suggest that the collagen-based casings behave as a better barrier to PAHs. PMID- 15895606 TI - Comparison of contaminant and residue levels in organic and conventional milk and meat products from northern Italy. AB - Due to the growing interest in organic products, a comparison between the chemical safety of organic and conventional products was undertaken. Milk and meat were the products chosen for study. The parameters evaluated to assess chemical safety were organochlorine pesticides, polychlorinated biphenyls (PCBs), lead, cadmium and mycotoxin contamination. Pesticides and PCBs residues in both organic and conventional milk and meat were lower than legal limits. Lead and cadmium residues were very low and did not differ between organic and conventional products. However, aflatoxin M1 contamination in some but not all samples of organic milk was significantly higher than those of conventional milk, although factors other than organic production might be implicated. PMID- 15895607 TI - Stability of the insecticide cypermethrin during tomato processing and implications for endocrine activity. AB - The thermal and pH stabilities of cypermethrin during food processing were investigated using tomato as a model food system and high-performance liquid chromatography as the analytical method. Cypermethrin was thermally unstable in aqueous conditions, where the hydrolysis of the pesticide was accelerated by heat. The mean proportion remaining after heating cypermethrin in water for 10 min was 66%, falling to 27% after 1 h. Similarly, thermal processing of canned tomatoes caused cypermethrin to degrade, with remaining levels in the final product ranging from 30 to 60% of the original. Cypermethrin was unstable at extreme pHs, with acid hydrolysis occurring faster than alkaline hydrolysis in phosphate buffers. The acidity of tomato paste (pH 4.3) caused cypermethrin levels to decrease by 30% within 12 days at 5 degrees C. The studies indicate that cypermethrin residues are likely to degrade by hydrolysis during food processing, thus reducing the exposure of consumers to cypermethrin. 3 Phenoxybenzaldehyde, a hydrolysis breakdown product of cypermethrin, was detected in the tomato paste and from the heating of cypermethrin in water at 100 degrees C. There is concern that the risk of breakdown products in terms of endocrine activity is unknown since in vitro studies reported that cypermethrin breakdown products display endocrine activity. PMID- 15895608 TI - Elimination of pesticides in olive oil by refining using bleaching and deodorization. AB - The effects of the processing steps on some pesticides during physical refining of olive oil were determined. Oil samples were spiked with four types of pesticides, i.e. endosulfan (alpha-endosulfan, beta-endosulfan and endosulfan sulphate), simazine, oxifluorfen and diflufenican, and subjected to physical refining (bleaching and deodorization). The removal of contaminants by bleaching earths was compared using five types of commercial earths. Deodorization assays under laboratory (10 kg) and discontinuous pilot plant (200 kg) conditions at different temperatures and treatment times were carried out. Bleaching seemed effective only for the elimination of simazine. For removal of the other pesticides tested, a physical refining treatment was required at 240 degrees C in the deodorizing stage for different periods (1 h for diflufenican, oxifluorfen and alpha-endosulfan, 2 h for beta-endosulfan and 3 h for endosulfan sulphate). PMID- 15895609 TI - Investigation into the validity of extrapolation in setting maximum residue levels for pesticides in crops of similar morphology. AB - Field trials were initiated to investigate if extrapolation procedures, which were adopted to limit costs of pesticide registration for minor crops, are valid. Three pairs of crops of similar morphology; carrots/swedes, cauliflower/calabrese (broccoli) and French beans/edible-podded peas; were grown in parallel at four different geographical locations within the UK. The crops were treated with both systemic and non-systemic pesticides under maximum registered use conditions, i.e. the maximum permitted application rates and the minimum harvest intervals. Once mature, the crops were harvested and analysed for residues of the applied pesticides. The limits of quantification were in the range 0.005-0.02 mg kg(-1). Analysis of variance and bootstrap estimates showed that in general, the mean residue concentrations for the individual pesticides were significantly different between crop pairs grown on each site. Similarly, the mean residue concentrations of most of the pesticides in each crop across sites were significantly different. These findings demonstrate that the extrapolations of residue levels for most of the selected pesticide/crop combinations investigated; chlorfenvinphos and iprodione from carrots to swedes; carbendazim, chlorpyrifos, diflubenzuron and dimethoate from cauliflower to calabrese; and malathion, metalaxyl and pirimicarb from French beans to edible-podded peas; appear invalid. PMID- 15895610 TI - Persistence of fenitrothion in oranges and clementines after treatment with emulsifiable concentrate and microencapsulate formulations. AB - The rate of decline of fenitrothion residues was investigated in oranges and clementines after treatment with two different kinds of commercial formulations: emulsifiable concentrate (Afidina M) and microencapsulate (Fenitrocap and IPM 400). The study was performed on the fruit and leaves over 131 and 161 days for oranges, and over 78 and 86 days for clementines, respectively. In fruit, the experimental data showed a similar behaviour of the active ingredient for both kinds of commercial formulations. High mean levels of fenitrothion (between about 0.4 and 0.8 mg kg(-1)) were persistent for at least 75 days after treatment in oranges and 50 days in clementines, with statistically significant declines observed only at days 110 and 78, respectively. A rapid decline of fenitrothion levels was observed in orange and clementine leaves during the starting phase followed by a slower decrease during the later stage; the decline was more pronounced with the treatment of emulsifiable concentrates. These findings are indicative of a poor degradability of fenitrothion in citrus fruits, and suggest that repeated or uneven applications of the pesticide should be avoided in order to exclude the risk of exceeding the maximum residue level permitted by the current regulations. PMID- 15895611 TI - Determination of trichothecenes in duplicate diets of young children by capillary gas chromatography with mass spectrometric detection. AB - Trichothecenes are mycotoxins produced by several fungal genera, mainly Fusarium species, that can contaminate a wide range of cereals used for human and animal consumption. They are associated with various adverse health effects in animals and humans such as feed refusal, vomiting and immunotoxic effects. A method based on capillary gas chromatography with mass spectrometric detection was developed and validated in-house for the determination of nine trichothecenes in duplicate diets of young children. The trichothecenes were extracted from the sample matrix by water/ethanol (90/10). The extracts were cleaned by means of ChemElut and Mycosep columns. The cleaned extracts were evaporated to dryness and derivatized to trimethylsilyl ethers at room temperature. The residues were dissolved in iso octane and washed with water. The final extracts were analysed for trichothecenes by GC-MS. The response was linear in the range tested (1-10 microg kg(-1)). Recoveries for the trichothecenes were between 70 and 111%, with the exception of nivalenol, which had a low recovery (34%). The limit of quantification for all trichothecenes was below 0.4 microg kg(-1). Seventy-four food samples from young children collected by 74 respondents in a duplicate diet study were analysed for trichothecenes with the developed method. The mean levels of deoxynivalenol, nivalenol, HT-2 toxin and T-2 toxin were 5.8, 0.3, 0.3 and 0.1 microg kg(-1), respectively. Based on the individual results, dietary intake calculations were made. For deoxynivalenol, the tolerable daily intake of 1 microg kg(-1) body weight was exceeded by nine respondents. For the combined intake of T-2 and HT-2 toxin, the temporary tolerable daily intake of 0.06 microg kg(-1) body weight was exceeded by nine respondents. PMID- 15895612 TI - Consumers' ability to discriminate aflatoxin-contaminated Brazil nuts. AB - The objectives of the study were to investigate the extent to which consumers can separate nuts with a high content of aflatoxin from sound nuts, and whether sorting results can be improved by information or whether they are affected by certain factors. A test panel consisting of 100 subjects was asked to crack 300 g Brazil nuts and to sort the nuts into those they considered edible and inedible. The test showed that consumers can, on current behaviour, discriminate aflatoxin contaminated Brazil nuts to a significant extent. The median and the 95th percentile of the total concentrations of aflatoxins (B1, B2, G1, G2) in the samples before sorting were 1.4 and 557 microg kg(-1), respectively, and in the edible fractions after sorting 0.4 and 56 microg kg(-1), respectively. Given that levels of aflatoxins before sorting exceed either 2 microg aflatoxin B1 kg(-1) or totally 4 microg aflatoxins kg(-1), there was no effect of aflatoxin concentrations before sorting on the probability of exceeding these thresholds in the edible fraction. This means that similar sorting results were obtained for samples with aflatoxin levels exceeding either of the two thresholds, irrespective of if the thresholds were exceeded with a few microg kg(-1) or up to more than 1000 microg kg(-1). None of the tested factors (such as sex, age, level of education, ethnic background or knowledge of mycotoxins) had any effects on the probability of exceeding either of the two aflatoxin thresholds. PMID- 15895613 TI - Bisphenol A in canned foods in New Zealand: an exposure assessment. AB - Exposure to bisphenol A (BPA) from the consumption of canned and bottled food has been determined for New Zealand adults. Eighty different canned foods purchased from retail outlets in Christchurch, New Zealand, between November 2003 and February 2004 were analysed for BPA concentration by gas chromatography/mass spectrometry. BPA was detected in all foods analysed except for soft drinks. Concentrations ranged from < 10 to 29 microg kg(-1), except for individual samples of tuna, corned beef and coconut cream, which were 109, 98 and 191 microg kg(-1) , respectively. The limit of quantitation was <10 microg kg(-1) for foods of low fat content (< 1%) and <20 microg kg(-1) for foods containing >1% fat. Mean concentration data were combined with 24-h dietary recall information for 4399 individual consumers. Mean and maximum exposures were 0.008 and 0.29 microg kg(-1) bw day(-1), respectively, well below the temporary tolerable daily intake of 10 microg kg(-1) bw day(-1) given by the European Commission in 2002. The results of the present survey suggest that the levels of BPA identified in canned foods are unlikely to be of concern to adult health, and there is no reason for consumers to change their consumption patterns as a result of these findings. When the concentration data found in the current survey are applied to an oestrogenicity model for an adult male, the contribution of BPA to the total oestrogenicity from 16 food components is 7%. The impact of this level of oestrogenicity remains unclear. PMID- 15895614 TI - Evaluation of migration models that might be used in support of regulations for food-contact plastics. AB - Materials and articles intended to come into contact with food must be shown to be safe because they might interact with food during processing, storage and the transportation of foodstuffs. Framework Directive 89/109/EEC and its related specific Directives provide this safety basis for the protection of the consumer against inadmissible chemical contamination from food-contact materials. Recently, the European Commission charged an international group of experts to demonstrate that migration modelling can be regarded as a valid and reliable tool to calculate 'reasonable worst-case' migration rates from the most important food contact plastics into the European Union official food simulants. The paper summarizes the main steps followed to build up and validate a migration estimation model that can be used, for a series of plastic food-contact materials and migrants, for regulatory purposes. Analytical solutions of the diffusion equation in conjunction with an 'upper limit' equation for the migrant diffusion coefficient, D(P), and the use of 'worst case' partitioning coefficients K(P,F) were used in the migration model. The results obtained were then validated, at a confidence level of 95%, by comparison with the available experimental evidence. The successful accomplishment of the goals of this project is reflected by the fact that in Directive 2002/72/EC, the European Commission included the mathematical modelling as an alternative tool to determine migration rates for compliance purposes. PMID- 15895615 TI - What to say to a skeptical metaphysician: a defense manual for cognitive and behavioral scientists. AB - A wave of recent work in metaphysics seeks to undermine the anti-reductionist, functionalist consensus of the past few decades in cognitive science and philosophy of mind. That consensus apparently legitimated a focus on what systems do, without necessarily and always requiring attention to the details of how systems are constituted. The new metaphysical challenge contends that many states and processes referred to by functionalist cognitive scientists are epiphenomenal. It further contends that the problem lies in functionalism itself, and that, to save the causal significance of mind, it is necessary to re-embrace reductionism. We argue that the prescribed return to reductionism would be disastrous for the cognitive and behavioral sciences, requiring the dismantling of most existing achievements and placing intolerable restrictions on further work. However, this argument fails to answer the metaphysical challenge on its own terms. We meet that challenge by going on to argue that the new metaphysical skepticism about functionalist cognitive science depends on reifying two distinct notions of causality (one primarily scientific, the other metaphysical), then equivocating between them. When the different notions of causality are properly distinguished, it is clear that functionalism is in no serious philosophical trouble, and that we need not choose between reducing minds or finding them causally impotent. The metaphysical challenge to functionalism relies, in particular, on a naive and inaccurate conception of the practice of physics, and the relationship between physics and metaphysics. PMID- 15895616 TI - Precis of the illusion of conscious will. AB - The experience of conscious will is the feeling that we are doing things. This feeling occurs for many things we do, conveying to us again and again the sense that we consciously cause our actions. But the feeling may not be a true reading of what is happening in our minds, brains, and bodies as our actions are produced. The feeling of conscious will can be fooled. This happens in clinical disorders such as alien hand syndrome, dissociative identity disorder, and schizophrenic auditory hallucinations. And in people without disorders, phenomena such as hypnosis, automatic writing, Ouija board spelling, water dowsing, facilitated communication, speaking in tongues, spirit possession, and trance channeling also illustrate anomalies of will--cases when actions occur without will or will occurs without action. This book brings these cases together with research evidence from laboratories in psychology to explore a theory of apparent mental causation. According to this theory, when a thought appears in consciousness just prior to an action, is consistent with the action, and appears exclusive of salient alternative causes of the action, we experience conscious will and ascribe authorship to ourselves for the action. Experiences of conscious will thus arise from processes whereby the mind interprets itself--not from processes whereby mind creates action. Conscious will, in this view, is an indication that we think we have caused an action, not a revelation of the causal sequence by which the action was produced. PMID- 15895617 TI - Grey matters. PMID- 15895618 TI - Listen to the people. PMID- 15895619 TI - Patients as experts. PMID- 15895620 TI - A role model for us all. PMID- 15895621 TI - Patients accelerating change. PMID- 15895622 TI - Evaluating the role of modern matrons. PMID- 15895623 TI - Implementing electronic integrated care pathways: learning from experience. PMID- 15895624 TI - Powerful relationships. PMID- 15895625 TI - The original spin doctors--the meeting of perception and insanity. PMID- 15895626 TI - Glowing greys and surface-white: the photo-geometric factors of luminosity perception. AB - The perception of luminosity is thought to depend upon the intensity of the stimulus: a surface begins to appear self-luminous when it emits or reflects a certain amount of light. This is known as the luminosity threshold. It is a common opinion among vision scientists that such a threshold is correlated to the intensity of a perceptually white surface, in the sense that only an area of the visual field with luminance higher than perceived surface-white will appear luminous. Here we show grey colours that appear luminous in virtue of surrounding luminance ramps. These ramps are intended to mimic halos seen around light sources in natural environments. The results of three experiments indicate that the phenomenon is in direct contradiction to the aforementioned assumptions and suggest the existence of separate perceptual pathways for self-luminosity perception and for surface-colour perception. PMID- 15895627 TI - The influence of illumination direction on the pictorial reliefs of Lambertian surfaces. AB - In order to assess the influence of illumination direction on shape constancy, we studied the pictorial relief of computer images of globular 3-D objects. We used two globally convex objects, one with a furrow and one with a dimple. Observers adjusted local surface attitude probes at 200-250 different locations in the image such that they seemed to lie on the pictorial surface. We manipulated the viewing direction and the illumination direction in a 2 x 2 orthogonal design. Viewing directions were chosen such that the image contained only a few, or no, contour singularities. Changes in the illumination direction were found to induce systematic changes in the settings for both viewing directions. Effects were especially pronounced for images that had no contour singularities. The results showed that a change in the illumination direction can change the local shape of the pictorial relief in addition to the bas-relief ambiguities of scaling and shearing in depth. We found that concavities in the pictorial relief are associated with the darker areas in the image. The deviation from shape constancy cannot be explained by bas-relief ambiguity since the required transformation between the shapes is nonlinear. PMID- 15895628 TI - Evidence for the correcting-mechanism explanation of the Kanizsa amodal shrinkage. AB - An object phenomenally shrinks in its horizontal dimension when shown on a 2-D plane as if the central portion of the object were partially occluded by another vertical one in 3-D space (the Kanizsa amodal shrinkage). We examined the predictions of the correcting-mechanism hypothesis proposed by Ohtsuka and Ono (2002, Proceedings of SPIE 4864 167-174), which states that an inappropriate operation of the mechanism that corrects a phenomenal increase in monocularly visible areas accompanied by a stereoscopic occluder gives rise to the illusion. In this study we measured the perceived width (or height in experiment 3) of a square seen behind a rectangle, while controlling other factors which potentially influence the illusion, such as the division of space or depth stratification. The results of five experiments showed that (a) the perceived width was not influenced when the occluder had a relatively large binocular disparity, but was underestimated when the occluder did not have disparity, and (b) the shrinkage diminished when the foreground rectangle was transparent, was horizontally oriented, or contained no pictorial occlusion cues. These results support the hypothesis that the correcting mechanism, triggered by pictorial occlusion cues, contributes to the Kanizsa shrinkage. PMID- 15895629 TI - Effect of attention on the detection and identification of masked spatial patterns. AB - The effect of attention on the detection and identification of vertically and horizontally oriented Gabor patterns in the condition of simultaneous masking with obliquely oriented Gabors was studied. Attention was manipulated by varying the set size in a visual-search experiment. In the first experiment, small target Gabors were presented on the background of larger masking Gabors. In the detection task, the effect of set size was as predicted by unlimited-capacity signal detection theory. In the orientation identification task, increasing the set size from 1 to 8 resulted in a much larger decline in performance. The results of the additional experiments suggest that attention can reduce the crowding effect of maskers. PMID- 15895630 TI - Attractiveness of own-race, other-race, and mixed-race faces. AB - Averaged face composites, which represent the central tendency of a familiar population of faces, are attractive. If this prototypicality contributes to their appeal, then averaged composites should be more attractive when their component faces come from a familiar, own-race population than when they come from a less familiar, other-race population. We compared the attractiveness of own-race composites, other-race composites, and mixed-race composites (where the component faces were from both races). In experiment 1, Caucasian participants rated own race composites as more attractive than other-race composites, but only for male faces. However, mixed-race (Caucasian/Japanese) composites were significantly more attractive than own-race composites, particularly for the opposite sex. In experiment 2, Caucasian and Japanese participants living in Australia and Japan, respectively, selected the most attractive face from a continuum with exaggerated Caucasian characteristics at one end and exaggerated Japanese characteristics at the other, with intervening images including a Caucasian averaged composite, a mixed-race averaged composite, and a Japanese averaged composite. The most attractive face was, again, a mixed-race composite, for both Caucasian and Japanese participants. In experiment 3, Caucasian participants rated individual Eurasian faces as significantly more attractive than either Caucasian or Asian faces. Similar results were obtained with composites. Eurasian faces and composites were also rated as healthier than Caucasian or Asian faces and composites, respectively. These results suggest that signs of health may be more important than prototypicality in making average faces attractive. PMID- 15895631 TI - Visual analysis of changes of motion in reaction-time tasks. AB - Subjects observed a random-dot pattern moving uniformly in the vertical direction (vector V1). The motion vector abruptly changed to V2, both in speed and direction simultaneously. It was found that the time of simple reaction to such changes V1 --> V2 can be described by a function of a single variable, [w(V1 - V2c) + (1 - w) V2N], 0 < w < 0.5, where V2c and V2N are the components of V2 collinear with and normal to V1. The choice-reaction time for changes in direction that are accompanied by changes in speed can be described by a function solely of the absolute value of V2N. Unlike the simple-reaction time, the choice reaction time was independent of the initial speed of motion. The processes that may be engaged in simple and choice reactions to motion are discussed. PMID- 15895632 TI - The trajectory effect in intermodal temporal order judgments. AB - Subjects judged which one of two patterns, a visual or a tactile pattern, had been presented first. The visual and tactile displays were placed in close spatial proximity. The patterns appeared to move across their respective displays. Although irrelevant to the temporal order judgment (TOJ), the direction of motion of the patterns--the trajectory--affected the judgments. When the leading pattern was moving towards the trailing pattern (consistent movement), subjects tended to judge it, correctly, as leading. When the leading pattern was moving away from the trailing pattern (inconsistent movement), subjects tended to judge it, incorrectly, as trailing. Changing the spatial position of the arrays such that the pattern trajectories were no longer towards one another eliminated the effect of movement on TOJs. Although there was a substantial difference in performance on consistent and inconsistent trials, there were no differences in subjects' ratings of their performances. The results demonstrate that the trajectory effect can be obtained multimodally. The issues whether the effect of motion alters the perceived temporal separation between the visual and tactile patterns, and whether the visual and tactile patterns are represented by a common framework, are discussed. PMID- 15895633 TI - Auditory saltation in the vertical midsagittal plane. AB - Auditory saltation is an illusion in which a train of clicks, the first half of which is presented at one location and the other half of which is presented from a second location, is perceived as originating not only from the anchor points, but also from locations between them. That is, intermediate members of the series of clicks have their spatial locations systematically misperceived. In the present study, auditory saltation was examined for the first time in the vertical midsagittal plane. Subjects rated the perceived continuity of motion for 8-click trains systematically varied in inter-click interval (ICI), direction of motion (up, down), and trial type ('saltation' versus 'real' motion). In all listeners, saltation stimuli supported robust saltation, but only for trials with ICIs less than about 120 ms. Real motion was rated as continuous for all ICIs. These data indicate that the auditory-saltation illusion can exploit monaural stimulus cues for source location in the generation of the illusory motion percept. PMID- 15895634 TI - Taking chances: risk assessment and management in a risk obsessed society. AB - Two earlier contributions to this journal on the topic of 'dangerous behaviour' are updated in the light of recent theoretical and practical developments in the assessment and management of risk in criminal justice and psychiatry. Such assessment and management need to be viewed against the background of current over-preoccupation with avoiding the hazards of daily living. PMID- 15895635 TI - Diaphragmatic pathology: a cause of clinically unexplained death in the perinatal/paediatric age group. AB - Sudden unexpected death in infancy and childhood requires a 'full' post-mortem investigation. Guidance from the Royal College of Pathologists recommends sampling of all the major organs. However, the diaphragm does not feature in this or in most lists of routine histology. Our aim is to emphasize the importance of sampling the diaphragm for histological examination during autopsy. We describe three autopsy cases of clinically unexplained death in the perinatal and paediatric age group that showed significant pathology of the diaphragm. In Case 1, a previously healthy five-year-old girl collapsed suddenly and died four days later. In Case 2, an eight-month-old infant had repeated episodes of respiratory arrest that culminated in death. Autopsy demonstrated a predominantly diaphragmatic myositis. In Case 3 a female neonate had a respiratory arrest three days after birth and died less than a month later. Autopsy showed multiple large calcified necrotic fibres in the diaphragm. The diaphragm is seldom sampled at autopsy. In the first two cases a predominantly diaphragmatic myositis was either the direct or underlying cause of death. In the third case long-standing diaphragmatic pathology of uncertain cause may have contributed to the original respiratory arrest. Had the diaphragm not been examined histologically, the cause of death would have remained unascertained in these cases. In cases of sudden death in infancy and childhood, failure to reach a diagnosis may lead to undue suspicion falling upon the child's carers. This underscores the need for full histology at post-mortem in child deaths, including diaphragmatic sampling. PMID- 15895636 TI - The impact of trends in psychotropic prescribing on the method of suicide in the elderly. AB - Suicide rates in the elderly have declined in many countries in recent years. This decline has been reported to be associated with increased prescribing of antidepressants, particularly selective serotonin reuptake inhibitors (SSRIs), antipsychotics and antimanic drugs and reduced prescribing of barbiturates, hypnotics and sedatives. This study examined the relationship between prescribing patterns of individual psychotropic drugs and suicide rates by specific methods of elderly suicides. There was a negative correlation between the prescription of tricyclic antidepressants, selective serotonin reuptake inhibitors, antipsychotics, antimanic drugs and non-opiate analgesics and a decline in elderly suicide rates due to poisoning by solid and liquid substances, hanging, strangulation and suffocation, drowning, firearms and explosives, and jumping from high places. There was a positive correlation between the prescription of barbiturates, hypnotics and sedatives and elderly suicide rates due to poisoning by solid and liquid substances, hanging, strangulation and suffocation, drowning, firearms and explosives, and jumping from high places. This study demonstrated that changes in prescribing patterns of individual psychotropic drugs do influence elderly suicide rates of the commonly used methods of suicide and suggest that this may be due to more accurate diagnostic-specific prescribing of psychotropic drugs. PMID- 15895637 TI - A postgraduate sexual assault forensic medicine program: sexual assault medicine from scratch. AB - A government-funded service to provide forensic and medical care to survivors of sexual assault was established in the Australian Capital Territory (ACT) in 2001. Doctors employed by the service lacked the specific skills required to care comprehensively for survivors. Our aim was to develop, implement and evaluate a sexual assault medical education program. It consisted of an 'in-house' education program, and external university course and incorporated team-building, networking activities and protocol development. Core elements were: forensic evidence collection, assessment and management of injuries, prevention of sexually transmissible infections and pregnancy, counselling and emotional support. Participant satisfaction and knowledge acquisition were evaluated using a semi-structured interview and a questionnaire. Seven doctors participated in a 16-session program conducted by the director and nurse coordinator with help from local forensic, legal and medical experts. All doctors successfully completed the Certificate in Forensic Medicine, and reported satisfaction with the program and their increased knowledge, particularly associated with collection of forensic evidence and court procedures. A compete set of protocols was developed and cohesive networks established. We have designed an effective education program for doctors working in the field of sexual assault and offer it as a template to other health professionals working in this area. PMID- 15895638 TI - Cross-sectional survey of the use of section 5(2), Mental Health Act 1983, within a psychiatric hospital. AB - A survey of the use of section 5(2), Mental Health Act 1983 within a psychiatric hospital was carried out between January and December 2001. Using computer-based patient records, it was found that 89 patients were placed on a section 5(2) between January and December 2001. Data on patient demographics, hospital admission date, date of implementation of section 5(2), date of reassessment after section 5(2) and outcome of the assessment was obtained from these records. In addition, the medical case notes of those patients identified (total of 81 case notes obtained) were studied with regard to the reasons for use of section 5(2), the process of implementing the section, the documentation of reassessments and the notification to the patient of his/her rights whilst under section. It was found that the conversion rates to a further section after implementation of section 5(2) were significantly higher amongst patients who are over 65 years than amongst patients under the age of 65 years (Yates corrected chi-square test 3.87, p=0.04.df=1. Risk ratio 0.68 (0.51-0.92, 95% CI). 53% of patients detained under section 5(2) were male, 47% were female. Sixty-four per cent of section 5(2)s were converted to a further section, 37% to a section 2 and 27% to a section 3. A longer period of time until reassessment of the patients led to a lower conversion rate to a further section. Fifty-four per cent of section 5(2)s were implemented within 24 hours of the patient's admission. Many current areas of practice do not meet with standards recommended by the Code of Practice. Higher conversion rates of section 5(2) to a further section may imply increasing psychiatric inpatient morbidity, hence higher numbers of patients being eligible for detention under the Mental Health Act 1983. PMID- 15895639 TI - Study of the relationship between a person's stature and the height of an ear imprint from the floor. AB - Occasionally ear prints are found at crime scenes. The height of the ear imprint may provide the police with information regarding the stature of the perpetrator and may therefore help to narrow down the number of suspects. The research provides calculations for the determination of stature from the height of the tragus imprint found at crime scenes. It takes into account various variables such as age, stature and gender. PMID- 15895640 TI - Patients with affective disorders admitted to maximum secure care (1999--2003). AB - The State Hospital, Carstairs, is a special secure psychiatric facility for Scotland and Northern Ireland. This study describes the background, illness and offence (where appropriate) characteristics of 25 patients admitted between 1999 and 2003 with a diagnosis of any affective disorder. Patients were, on average, 37 years old and had a lengthy history of psychiatric contact. Patients with an affective disorder were more likely to be admitted under civil proceedings, following a transfer from hospital, as opposed to being admitted under criminal procedure. Sixty per cent were discharged to prison, court or local hospitals within one year of their admission to special secure care. Significant violence during an episode of mania or hypomania (even in the presence of psychotic symptoms) appears rare. Recurrent unipolar depression resistant to first and second line treatments in women, and lithium withdrawal mania in male bipolar patients were the most common clinical problems. Treatments aimed at reducing co morbid substance misuse, and improving compliance with mood stabilisers, appear important in managing this patient group. PMID- 15895641 TI - Needs assessment in forensic learning disability. AB - This paper gives the results of a needs assessment of a group of learning disabled forensic patients from two Strategic Health Authority areas in the north of England. The patients were found to be a heterogeneous group with wide-ranging psychiatric needs. The majority were cared for outside their geographical area of origin, either in specialist NHS facilities or the independent sector. Those with an additional diagnosis of mental illness were most likely to be detained in NHS facilities within the region: a diagnosis of personality disorder was associated with placement in either a high secure setting or the independent sector. Individuals with a clinical diagnosis of mental retardation were most likely to be detained in services provided by specialist learning disability/mental health trusts out of area. There was a small group of females who were all placed outside the region. Offending behaviour was most likely to consist of violence against the person, sexual offences and arson. The majority assessed were felt to have long term needs. The study raised important implications for future provision of forensic services in the area, particularly the need to offer services with treatment programmes tailored to the needs of the population under review. PMID- 15895642 TI - Is there a need for elderly forensic psychiatric services? AB - The population of the elderly in most developed nations is on the increase. Furthermore, the prevalence of mental disorder amongst elderly offenders is high. The true extent of 'elderly' crime is unknown because much of it goes undetected and unreported. This leads to a failure to detect mental illness in such offenders. Court diversion schemes may improve recognition of mental illness but these schemes usually tend to deal with the more severe crimes. This may result in an overestimation of the amount of serious crime committed by the elderly and a failure to detect mental illness amongst those who commit less serious crimes. Efforts to service this hidden morbidity call for multi-agency collaboration. Improved detection and reporting of crimes is essential if mental health difficulties in the elderly are not to go unnoticed. The needs of elderly mentally-disordered offenders are complex and fall within the expertise of old age and forensic psychiatry, without being adequately met by either one. Therefore, consideration should be given to the development of a tertiary specialist forensic old-age psychiatry service. PMID- 15895643 TI - Dowry--a deep-rooted cause of violence against women in India. AB - The world has entered the new millenium but it is a harsh reality that the woman in India has long been ill-treated in our male-dominated society. She is deprived of her independent identity and is looked upon as a commodity. She is not only robbed of her dignity and pride by way of seduction by the men outside, but also may become a victim of cruelty by her saviours, within the four walls of her own house. However, her trauma does not end here, it may even go to the extent of forcing her to commit suicide or she may be burnt to death for various reasons, including that of dowry. This type of violence transgresses the boundaries of caste, class, region or religion and is prevalent in almost all societies in India. The system of dowry is a social practice which on its own has claimed the lives of scores of women--both young and old, and has made life a virtual hell for many more. Unfortunately, education among women has not produced a reformative effect on their social outlook, nor encouraged any change in them conducive to social upliftment. Those parents who prefer not to take dowry for their well-qualified and settled male children, are in fact considered 'strange' by the society and doubts about 'the respectability of the groom's family' are usually raised. The present study makes an in-depth review of the dowry system in India and analyses the reasons which have demonized dowry into its present commercialized and institutionalized form. PMID- 15895644 TI - Fatal occupational injuries in the South Delhi construction industry: a retrospective study. AB - One hundred and forty-five unselected autopsy cases of construction site accidents received from South Delhi were studied during the period from 1996- 2002. Data for the study was gathered from autopsy reports and hospital records. The cases represented approximately 1.61% of all autopsy cases received from South Delhi at the All India Institute of Medical Sciences, New Delhi (India). Data was analysed with regard to the age and sex of the victim, the part of the body involved, the manner of accident, the cause of death and the pattern of injuries in different body regions. Death occurred at the scene of the fatal event in thirty-four cases; forty-three cases were dead on arrival at the hospital; sixty-eight cases died after being admitted to the hospital. Ethanol was detected in the blood of 16% of the cases. PMID- 15895645 TI - 1. An uncommon case of zoophilia: a case report. AB - Humans show a wide array of sexual preferences and behaviour. Zoophilia, or bestiality, consists of sexual intercourse by a human being with a lower animal. There is a paucity of literature on zoophilia. We describe a rare case of preferential zoophilia involving multiple animals. The perpetrator developed this habit only during episodes of depression. It is recommended that due to the apparent common occurrence of this condition, questions exploring a topic which appears to be routinely ignored should be included in the psychiatric interview. PMID- 15895646 TI - 2. An unusual death due to the impalement of a gear stick into the brain stem through the nasal cavity: a case report. AB - A 59-year-old male was driving a car on the road and was involved in a traffic accident, colliding with a tanker and a big lorry. When an ambulance arrived at the scene, he was in a state of cardiopulmonary arrest and was bleeding profusely from his right nostril. He was confirmed dead at hospital. The autopsy showed a laceration of the right upper lip extending to the right nostril. In the basal skull there was a notable perforation at the ethmoid bone together with the central part of the sphenoid bone including the sella turcica. In accordance with the basal skull bone fractures, there were pronounced contusion injuries at the brain stem and a contusion injury was also observed in the right part of the cerebellum. After careful investigation of a causative stick-like item that was present inside the car, it was concluded that a severe movement of the man's body, as a result of the traffic collision, caused the gear stick which was fixed to the steering wheel to become impaled in the man's right nostril. The gear stick passed through the nasal cavity and into the basal skull bones, resulting in fatal brain stem injuries. PMID- 15895647 TI - 3. Sudden death due to pulmonary thromboembolism after car driving: a case report. AB - An autopsy case of sudden death due to pulmonary thromboembolism after a car trip is reported. A 56-year-old Japanese man with hypertension and atrial fibrillation suddenly died immediately after having driven for two and a half hours. At autopsy, the pulmonary arteries were found to be filled with dark-red, coiling thrombi consisting of fibrin and red blood cells. In the dilated right popliteal vein, a thrombus consisting of fibroblasts, a large number of collagen fibres, and newly formed capillaries was attached to the vessel wall. The cause of death was determined to be pulmonary embolism due to the thrombus of the right popliteal vein. Two and a half hours of prolonged sitting while driving a car encouraged thrombus formation. We believe that this case, the first autopsy case of fatal pulmonary thromboembolism after a car trip to be reported, highlights the importance of predicting venous thrombosis of the leg or fatal venous thromboembolism after a car trip. PMID- 15895648 TI - Shaken-baby syndrome: a challenging diagnosis. PMID- 15895649 TI - The semantics of disability. PMID- 15895650 TI - Private practice Southern style. PMID- 15895651 TI - Breaking barriers. PMID- 15895652 TI - The integrated future. PMID- 15895653 TI - A mixed bag. PMID- 15895654 TI - Stealing the scene. PMID- 15895656 TI - Easing patient discomfort. PMID- 15895655 TI - Focus on the family. PMID- 15895657 TI - Toward the Tibetan plateau. PMID- 15895658 TI - Uncertain times change health care focus. PMID- 15895659 TI - A pen to remember. PMID- 15895660 TI - Rehab and the bariatric patient. PMID- 15895661 TI - Successful aging. PMID- 15895662 TI - The power of prevention. PMID- 15895663 TI - Using surface electromyography. PMID- 15895664 TI - MJ Care never gives up. PMID- 15895665 TI - Carving out a niche. PMID- 15895666 TI - Polio returns from the dead. PMID- 15895667 TI - The outsourcing option. PMID- 15895668 TI - Aortic arch and descending thoracic aortic aneurysms: experience with stent grafting for second-stage "elephant trunk" repair. AB - Reconstruction of aortic arch and descending thoracic aortic aneurysms (TAAs) is technically challenging and associated with significant morbidity and mortality. We report our experience with extensive TAAs using a two-stage "elephant trunk" repair, with the second stage completed using an endovascular stent graft (ESG). Over 6 years, 111 patients underwent ESG treatment of TAAs at Mount Sinai Medical Center. Twelve of these patients were referred for ESG placement for the second stage of elephant trunk reconstruction because comorbidities placed them at high risk of open surgical repair. Our database was analyzed for technical and clinical success and perioperative complications. The mean follow-up was 11.8 months (range 1-64 months). Twelve patients (five women and seven men) with a mean age of 69 +/- 10 years underwent repair of their distal aortic arch and descending TAAs. These aneurysms included nine atherosclerotic aneurysms, one pseudoaneurysm, and two penetrating atherosclerotic ulcers. Three patients were symptomatic. Stent graft repair was technically successful in 91.7% or 11 of 12 patients. Excessive aortic arch tortuosity resulted in failure to deploy a stent graft in one patient. An antegrade approach through the open elephant trunk was used in two patients with severe iliac occlusive disease. Endoleaks (type 2) were identified in two patients with no aneurysm expansion; however, a 14 mm expansion over 1 year occurred in a patient with no identifiable endoleak. One early mortality occurred in a patient with a ruptured 6 cm infrarenal AAA after successful exclusion of the 8 cm TAA. Second-stage elephant trunk reconstruction of an extensive TAA using an ESG is effective in the short term. Its long-term durability remains to be determined. PMID- 15895669 TI - Type I aneurysmosis: complementary index for diagnosis. AB - The aim of this study was to investigate the diameters of the superficial femoral artery (SFA) and the popliteal artery (PA) in patients with type I aneurysmosis (diffuse aneurysm in the aortoiliac-common femoral segments combined with arteriomegaly of the femoropopliteal segments). The ratio between these diameters (SFA/PA index) was calculated. A retrospective study was carried out on 76 limbs of 38 patients with type I aneurysmosis. The SFA/PA index calculated on these patients was compared with the index of a control group: 76 limbs of 38 patients with abdominal aortic aneurysm. The SFA/PA index proved to be <1.00 in the limbs of patients with type I aneurysmosis, whereas it was > or =1.00 in all patients with an abdominal aortic aneurysm. There was a statistically significant difference between the type I aneurysmosis group and the control group (p < .001). This finding can be considered a complementary sign in the diagnosis of type I aneurysmosis. PMID- 15895670 TI - Use of proximal aortic cuffs as an adjunctive procedure during endovascular aortic aneurysm repair. AB - The purpose of this study was to evaluate the incidence and durability of additional proximal cuffs during endovascular abdominal aortic aneurysm repair (EVAR). A retrospective review of 90 EVAR patients was conducted. Postoperative survival, proximal sealing zone-related complications, and secondary procedures were analyzed. Additional proximal cuffs were used in 11%. Their use did not affect postoperative survival (p = .58), type I endoleak rate (4.4%; p = .19), or the need for sealing zone-related secondary procedures (6.3%; p = .38) compared with patients without cuff placement but was related to a higher cumulative graft migration rate (2.2% overall p = .02). Two patients (2.5%; p = .79) underwent conversion to open surgery, both for proximal sealing zone-related complications. Application of proximal cuffs appears to be an effective intraoperative adjunctive procedure to achieve a proximal seal during EVAR, with favorable midterm results. However, the risk of late endograft migrations may be elevated in this group. PMID- 15895671 TI - Selective shunting during carotid endarterectomy. AB - Carotid endarterectomy (CEA) is an effective treatment for significant carotid atherosclerosis. Perioperative stroke, a devastating complication, may be partially circumvented by shunting. However, routine shunt use is not without complications and does not benefit every patient. Our study is designed to determine whether CEA under general anesthesia, without cerebral monitoring, can be safely done with shunting only in the presence of poor internal carotid artery back-bleeding or contralateral carotid occlusion or critical stenosis. The medical records of 995 carotid operations were reviewed. A subset of 117 operations was performed on 112 patients using selective shunting. Data were analyzed and outcomes compared. For the selective shunt group, indications for redo operations (n=13) were recurrent asymptomatic high-grade stenosis in 69% and amaurosis fugax or transient ischemic attack in 31%. Indications for primary CEA (n=104) were asymptomatic high-grade stenosis in 59%, amaurosis fugax or transient ischemic attack in 36%, previous stroke in 3%, and global ischemia in 2%. A selective shunt was used in 29% of all symptomatic and 11% of all asymptomatic patients. No cerebral monitoring was used. There were no perioperative deaths and no permanent cranial nerve injuries, and there was one stroke (0.8%) from postoperative carotid thrombosis in a shunted patient. The average length of stay was 1.6 days for the non-shunt group and 2.2 days for the shunt group. The routine shunt group (n=878) had an overall stroke rate of 0.7%, no permanent cranial nerve deficits, and a mean hospital stay of 2.6 days. CEA under general anesthesia with selective shunting can be performed safely without cerebral monitoring. PMID- 15895672 TI - Effect of ultrasonography in the assessment of carotid artery stenosis. AB - Although ultrasonography (US) advantageously portrays lumen and wall thickness, velocity criteria have been used primarily to interpret carotid artery stenosis. The relationship of US and velocity measurements was investigated. Peak-systolic and end-diastolic velocities (PSV, EDV) increase exponentially as the lumen of the internal carotid artery narrows and the percent stenosis (%S) increases. We tested the consistency of the, relationship between carotid velocities and US %S in two distinct data sets. One data set was used to obtain regression equations relating velocity parameters and %S based on US. Validation of these equations was conducted using a separate, independent data set. US measurements were classified in 12 %S intervals, PSV, EDV, the ratio of the internal carotid artery to the common carotid artery PSV, and %S were entered consecutively until 10 records for each %S interval were obtained. Regression equations obtained in the first data set were used to predict %S in the second data set. Predicted %S was then compared with actual US %S. The highest correlation in the first data set (r = .89) was between %S and the natural logarithm (In) of PSV. This In PSV -%S equation was then applied to a second data set of an additional 120 carotid duplex images. In the second data set, actual %S and PSV-predicted %S differed by >10% in 38 cases (32%). When all velocity-%S regression equations were used for comparison, differences between actual and at least one velocity-predicted %S were >10% in 19% of the arteries. Conversely, actual %S matched at least one prediction of %S based on velocity data in 81% of the cases. US %S differed significantly from single velocity-based estimates of %S in at least one-third of the cases. On the other hand, four of five US measurements were confirmed by at least one velocity parameter. Emphasis on US, in addition to velocity data, is recommended for the interpretation of duplex US carotid examinations. PMID- 15895673 TI - Role of apoptosis and proteolysis in the pathogenesis of iliac artery aneurysms. AB - The objective of this study was to investigate the role of inflammation, programmed cell death, its molecular modulators, and proteolysis in the pathogenesis of iliac artery aneurysms (IAAs). Nineteen IAA specimens were obtained from patients undergoing elective surgical repair. All were males with ages ranging from 55 to 85 years (mean 73 years). Controls were iliac arteries (n=6) retrieved from surgical patients without aneurysmal disease. Standard histochemical techniques were used to assess elastic lamellae fragmentation and inflammatory infiltrate in aneurysmal and normal tissues Identification of different types of cells in the aneurysm wall and detection of death-pro molecules, Fas, p53, perforin, apoptosis-mediating bcl-2 family proteins, apoptotic death substrate, and poly(adenosine diphosphate-ribose) polymerase were performed immunohistochemically. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated digoxigenin-deoxyuridine triphosphate nick end-labeling (TUNEL) assay and caspase activity. Proteolytic activity was determined by 10% gelatin gel zymography. There is a conspicuous disruption and fragmentation of elastic lamellae in IAAs compared with normal arteries. Increased gelatinolytic activity was observed at 92, 72, and 67 kDa in the aneurysmal tissues. There was a significant loss of vascular smooth muscle cells (VSMCs) in the IAA walls compared with normal arteries (p < .02). Large numbers of inflammatory cells were observed in the IAA specimens (p = .01). Only aneurysmal arteries showed CD8+ T cells expressing death-promoting molecules. CD3+, CD8+, CD20+, CD30+, and CD68+ immunoreactive cells were significantly more prominent in the aneurysmal tissues than in the control arteries. There was a significant increase in the number of cells undergoing apoptosis in aneurysmal tissue than in the normal vessels (p < .02), as well as in the expression of bax, p53, CPP-32, and Fas. Apoptotic cells and proapoptotic molecules predominantly localized to the inflammatory infiltrate. VSMC apoptosis was significant in IAAs. The data confirm the architectural disruption of the IAA wall and illustrate an apparent biologic response involving inflammatory infiltrate, apoptosis, and signaling molecules capable of initiating cell death. In addition to compromising the mechanical integrity of the vessel wall, VSMC loss may contribute to imbalance in the protein profile, accelerating extracellular matrix degradation that could favor IAA development. PMID- 15895674 TI - Effect of clopidogrel on platelet aggregation and intimal hyperplasia following carotid endarterectomy in the rat. AB - Intimal hyperplasia results in significant morbidity and mortality following vascular intervention. Both platelets and elevated homocysteine have been implicated in the development of intimal hyperplasia. We previously demonstrated that a locally applied antiplatelet agent decreases the development of intimal hyperplasia. We were therefore interested in a systemic antiplatelet agent, clopidogrel. We hypothesized that clopidogrel would decrease platelet aggregation and activity and intimal hyperplasia. Male Sprague-Dawley rats underwent carotid endarterectomy (CEA) and treatment with either placebo or varying regimens of clopidogrel, including chronic, pre-CEA bolus, chronic plus pre-CEA bolus, and chronic plus post-CEA bolus; a homocystine diet was used to elevate both plasma homocysteine and the degree of intimal hyperplasia. Platelet aggregation, platelet activity, and intimal hyperplasia were then assessed. Platelet aggregation was not decreased with chronic clopidogrel; however, it was decreased with pre-CEA bolus clopidogrel. Similarly, platelet activity was not inhibited by chronic clopidogrel but was inhibited by pre-CEA and chronic plus pre-CEA bolus clopidogrel. Neither chronic, pre-CEA bolus, chronic plus pre-CEA bolus, nor chronic plus post-CEA bolus clopidogrel resulted in a decrease in intimal hyperplasia. Although pre-CEA bolus clopidogrel resulted in a decrease in both platelet aggregation and activity, it was unable to decrease the development of intimal hyperplasia at any dose. Additional factors must therefore contribute to the pathologic development of intimal hyperplasia. PMID- 15895675 TI - Distal peripheral microembolism. AB - Distal peripheral microembolism is caused by embolization of atherosclerotic debris into small arteries and arterioles. The recent advances in endovascular technique have been met with a gradual increase in the incidence of iatrogenic atheroembolism. This review seeks to explore the nature of distal peripheral microembolism, pathophysiology, and the management options, with a focus on iatrogenic distal peripheral microembolism. PMID- 15895676 TI - Endovascular management of a gunshot wound injury to the innominate artery and brachiocephalic vein. AB - Surgical repair of penetrating injuries of the thoracic outlet with combined arterial and venous involvement is associated with considerable morbidity and mortality. A 37-year-old man presented to the emergency room with a left-sided penetrating zone I neck injury caused by a close-range handgun shot. This had resulted in an injury to the innominate artery and the origin of the right common carotid artery, with shunting to the brachiocephalic vein. This was managed endovascularly by stenting of the innominate artery and by coiling of the origin of the carotid artery. An endovascular approach to this injury is feasible and has the advantage of appropriate visualization of the vascular lesions with limited blood loss during the repair. PMID- 15895677 TI - Suicide inhibition of acetohydroxyacid synthase by hydroxypyruvate. AB - Acetohydroxyacid synthase (Ec 2.2.1.6) catalyses the thiamine diphosphate dependent reaction between two molecules of pyruvate yielding 2-acetolactacte and CO2. The enzyme will also utilise hydroxypyruvate with a k(cat) value that is 12% of that observed with pyruvate. When hydroxypyruvate is the substrate, the enzyme undergoes progressive inactivation with kinetics that are characteristic of suicide inhibition. It is proposed that the dihydroxyethyl-thiamine diphosphate intermediate can expel a hydroxide ion forming an enol that rearranges to a bound acetyl group. PMID- 15895678 TI - Quantitative structure activity relationships (QSAR) of substituded (S) phenylpiperidines as preferential dopamine autoreceptor antagonists. AB - A QSAR analysis for substituted (S)-phenylpiperidines as dopamine (DA) antagonists is described. The studied derivatives differ at the nitrogen substitutent (R) and at the substitutents (X) of the phenyl-ring. The analysis was done using the C-QSAR suite program (Biobyte) through the Internet. Clog P, CMR, M(vol), B1 and L (the Verloop's sterimol parameters for the substitutents) were used as parameters. In all the three studied cases clog P plays a significant part in the QSAR of DA antagonists, followed by the steric factors. In one case the electronic effect contributes significantly. PMID- 15895679 TI - Amidine derivatives and Leishmania amazonensis: an evaluation of the effect of nitric oxide (NO) production on the parasite-macrophage interaction. AB - Previous work has demonstrated that N-N'-diphenyl-R-benzamidine was highly effective against Leishmania amazonensis promastigotes/axenic amastigotes and Trypanosoma evansi trypomastigotes and the compound with a methoxy substituent, was the most effective derivative in the parasite-macrophage interaction. Comparative analysis of the nitric oxide (NO) released from the culture infection's supernatant showed the amidine to be less effective than pentamidine Isethionate as a reference drug. Additionally, in order to verify if the methoxylated derivative interferes with NO production by L. amazonensis, the effect of the amidine on the constitutive nitric oxide synthase (cNOS) purified from parasites, was examined, but demonstrated less activity in comparison with the reference drug. This data contributes to studies concerning the metabolic targets present in Leishmania parasites for leishmanicidal drugs. PMID- 15895680 TI - Evaluation of effect of some corticosteroids on glucose-6-phosphate dehydrogenase and comparative study of antioxidant enzyme activities. AB - Corticosteroids are anti-inflammatory drugs that are similar to the natural corticosteroid hormones produced by the cortex of the adrenal glands. The objective of this study was to scrutinize effects of some corticosteroids on glucose-6-phosphate dehydrogenase (G6PD) and some antioxidant enzymes. Initially, G6PD was purified from human erythrocytes by using ammonium sulphate precipitation and affinity chromatography. The two drugs, dexamethasone phosphate and prednisolone, investigated on the purified enzyme inhibited the enzyme activity. Comparative in vivo studies were performed to determine the effects of dexamethasone phosphate on the antioxidant enzyme activities using Spraque-Dawley rats. G6PD and catalase (CAT) activities were found significantly lower than in the control, whereas glutathione peroxidase (GP) activity was significantly increased in the erythrocytes of rats the receiving drug; glutathione reductase (GR) activity was unaffected. The results imply that dexamethasone phosphate may affect oxidative stress by changing antioxidant enzyme activities. PMID- 15895681 TI - Structural requirements for inhibitors of cytochromes P450 2B: assessment of the enzyme interaction with diamondoids. AB - The series of diamondoids: adamantane, diamantane, triamantane, 2-isopropenyl-2 methyladamantane and 3-isopropenyl-3-methyldiamantane (3-IPMDIA), were employed to elucidate the molecular basis of their interaction with the active site of cytochromes P450 (CYP) of a 2B subfamily. These potent inhibitors of CYP2B enzymes were docked into the homology model of CYP2B4. Apparent dissociation constants calculated for the complexes of CYP2B4 with docked diamandoids agreed closely with the experimental data showing inhibition potency of the compounds and their binding affinity to CYP2B4. Superimposed structures of docked diamondoids mapped binding site residues. As they are mainly non-polar residues, the hydrophobicity plays the major role in the binding of diamondoids. Overlapping structure of diamondoids defined an elliptical binding cavity (5.9A inner diameter, 7.9A length) forming an angle of approximately 43 degrees with the heme plane. CYP2B specific diamondoids, namely 3-IPMDIA, showing the highest binding affinity, should be considered for a potential clinical use. PMID- 15895682 TI - Effects of low molecular weight plasma inhibitors of rainbow trout (Oncorhynchus mykiss) on human erythrocyte carbonic anhydrase-II isozyme activity in vitro and rat erythrocytes in vivo. AB - The effects of low molecular weight plasma inhibitors from rainbow trout (Oncorhynchus mykiss) (RT) were investigated on the carbonic anhydrase enzyme (CA) activities in in vitro human and in in vivo Sprague-Dawley rat erythrocytes. The RT blood was used as extracellular fluid (plasma) source and plasma inhibitors were obtained by dialysis of the plasma. For the in vitro study, human carbonic anhydrase-II (HCA-II) isozyme was obtained by Sepharose 4B-L-tyrosine sulfanylamide affinity chromatography with an overall purification of about 646 fold. The enzyme (specific activity of 7750 EU/mg protein) was obtained with a yield of 71.1% and SDS-PAGE showed a single band. From in vitro studies, the I50 value for RT plasma inhibitors obtained was 0.37 mg/ml. From in vivo studies on rat erythrocytes, CA activity was significantly inhibited by the inhibitors from the extracellular fluid of RT for up to 3 h (p < 0.05) following intraperitoneal administration. PMID- 15895683 TI - Synthesis and inhibitory potential towards acetylcholinesterase, butyrylcholinesterase and lipoxygenase of some variably substituted chalcones. AB - A series of variably substituted chalcones were synthesized by condensation of substituted acetophenones with mono-, di- or trisubstituded benzaldehydes. It was observed that some of these compounds have the potential to inhibit acetylcholinesterase, whereas others show activity against butyrylcholinesterase, depending on the substitution pattern at the two aromatic rings of these chalcones. Similarly, lipoxygenase was inhibited by two of these compounds. It has been observed that inhibition of the three enzymes was concentration dependent with the IC50 values ranging from 28.2-134.5 microM against acetylcholinesterase, 16.0-23.1 microM against butyrylcholinesterase and 57.6 71.7 microM against lipoxygenase, respectively. PMID- 15895684 TI - Inhibition of porcine small intestinal sucrase by valienamine. AB - Valienamine, an aminocyclitol, has been isolated from the enzymolysis broth of validamycins. The absolute configuration of valienamine is similar to that of alpha-D-glucose. The inhibitory effect of this amino-sugar analog of alpha-D glucose, valienamine, on porcine small intestinal sucrase was examined. Valienamine was found to be potent, competitive reversible inhibitor of porcine small intestinal sucrase in vitro with an IC50 value of 1.17 x 10(-3)M. Valienamine also exhibited dose-dependent, instantaneous inhibition of porcine small intestinal sucrase. The inhibition of porcine small intestinal sucrase by valienamine was pH-independent. PMID- 15895685 TI - Syntheses and anti-inflammatory activity of diphenylamine-2,2'-dicarboxylic acid and its metal complexes. AB - Diphenylamine-2,2'-dicarboxylic acid and its Cu(II), Ni(II), Co(II) and Zn(II) complexes have been synthesized and characterized by their elemental analyses, molecular weight determination, molar conductance, infrared and electronic spectra and magnetic measurements. The Zinc complex was tested by different methods for its anti-inflammatory activity and found to be equipotent to naproxen and ibuprofen, though at higher doses. PMID- 15895686 TI - Glutamylsulfamoyladenosine and pyroglutamylsulfamoyladenosine are competitive inhibitors of E. coli glutamyl-tRNA synthetase. AB - 5'-O-[N-(L-glutamyl)-sulfamoyl] adenosine is a potent competitive inhibitor of E. coli glutamyl-tRNA synthetase with respect to glutamic acid (K(i) = 2.8 nM) and is the best inhibitor of this enzyme. It is a weaker inhibitor of mammalian glutamyl-tRNA synthetase (K(i) = 70 nM). The corresponding 5'-O-[N-(L pyroglutamyl)-sulfamoyl] adenosine is a weak inhibitor (K(i) = 15 microM) of the E. coli enzyme. PMID- 15895687 TI - Effects of some antibiotics on glutathione reductase activities from human erythrocytes in vitro and from rat erythrocytes in vivo. AB - The effects of streptomycin sulfate, gentamicin sulfate, thiamphenicol, penicillin G, teicoplanin, ampicillin, cefotaxime, and cefodizime on the enzyme activity of glutathione reductase (GR) were studied using human and rat erythrocyte GR enzymes in in vitro and in vivo studies, respectively. The enzyme was purified 5,342-fold from human erythrocytes in a yield of 29% with 50.75 U/mg. The purification procedure involved the preparation of hemolysate, ammonium sulfate precipitation, 2',5'-ADP Sepharose 4B affinity chromatography and Sephadex G-200 gel filtration chromatography. Purified enzyme was used in the in vitro studies, and rat erythrocyte hemolysate was used in the in vivo studies. In the in vitro studies, I50 and K(i) values were 12.179 mM and 6.5123 +/- 4.1139 mM for cefotaxime, and 1.682 mM and 0.7446 +/- 0.2216 mM for cefodizime, respectively, showing the inhibition effects on the purified enzyme. Inhibition types were noncompetitive for cefotaxime and competitive for cefodizime. In the in vivo studies, 300 mg/kg cefotaxime and 1000 mg/kg cefodizime when administered to rats inhibited enzyme activity during the first 2h (p < 0.01). Cefotaxime led to increased enzyme activity at 4h (p < 0.05), but neither cefotaxime nor cefodizime had any significant inhibition or activation effects over 6 h (p > 0.05). PMID- 15895688 TI - Purification and characterization of carbonic anhydrase from bovine stomach and effects of some known inhibitors on enzyme activity. AB - Carbonic anhydrase (CA) was purified from four different cell localisation (outer peripheral, cytosolic, inner peripheral and integral) in bovine stomach using affinity chromatography with Sepharose-4B-L-tyrosine sulphanilamide. During the purification steps, the activity of the enzyme was measured using p-nitrophenyl acetate at pH 7.4. Optimum pH and optimum temperature values for all CA samples were determined, and their K(m) and V(max) values for the same substrate by Lineweaver-Burk graphics. The extent of purification for all CA localizations was controlled by SDS-PAGE. The K(m) values at optimum pH and 20 degrees C were 0.625 mM, 0.541 mM, 0.785 mM and 0.862 mM with p-nitro phenyl acetate, for all CA localizations. The respective V(max) values at optimum pH and 20 degrees C were 0.875 micromol/L min, 0.186 micromol/L min, 0.214 micromol/L min and 0.253 micromol/L min with the same substrate. The K(i) and I50 values for the inhibitors sulphanilamide, KSCN, NaN3 and acetazolamide were determined for all the CA localizations. PMID- 15895689 TI - Organometallic-based antibacterial and antifungal compounds: transition metal complexes of 1,1'-diacetylferrocene-derived thiocarbohydrazone, carbohydrazone, thiosemicarbazone and semicarbazone. AB - Organometallic-based, 1,1'-diacetylferrocene-derived antibacterial and antifungal thiocarbohydrazone, carbohydrazone, thiosemicarbazone and semicarbazone have been prepared by condensing equimolar amount of 1,1'-diacetylferrocene with thiocarbohydrazide, carbohydrazide thiosemicarbazide and semicarbazide, respectively. These were used as ligands for the preparation of their cobalt (II), copper (II), nickel (II) and zinc (II) metal complexes. All the synthesized ligands and their complexes were characterized by IR, NMR, elemental analyses, molar conductances, magnetic moments and electronic spectral data. These synthesized compounds were screened for their antibacterial activity against Escherichia coli, Bacillus subtillis, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella typhi, and for antifungal activity against Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium solani and Candida glaberata using the agar-well diffusion method. All the compounds showed good antibacterial and antifungal activity which increased on coordination with the metal ions thus, introducing a novel class of organometallic-based antibacterial and antifungal agents. PMID- 15895690 TI - Inhibition by N'-nitrosonornicotine of the catalytic activity of glutamate dehydrogenase in alpha-ketoglutarate amination. AB - The effect of N'-nitrosonornicotine (NNN), one of the tobacco-specific nitrosamines, on the catalytic activity of glutamate dehydrogenase (GLDH) in the alpha-ketoglutarate amination, using reduced nicotinamide adenine dinucleotide as coenzyme, was studied by a chronoamperometric method. The maximum reaction rate of the enzyme-catalyzed reaction and the Michaelis-Menten constant, or the apparent Michaelis-Menten constant, were determined in the absence and presence of NNN. NNN remarkably inhibited the bio-catalysis activity of GLDH, and was a reversible competitive inhibitior with K(i), estimated as 199 micromol l(-1) at 25 degrees C and pH 8.0. PMID- 15895691 TI - Purification and characterization of a XIP-type endoxylanase inhibitor from rice (Oryza sativa). AB - A rice XIP-type inhibitor was purified by affinity chromatography with an immobilized Aspergillus aculeatus family 10 endoxylanase. Rice XIP is a monomeric protein, with a molecular mass of ca. 32 kDa and a pI of ca. 5.6. Its N-terminal amino acid sequence was identical to that of a rice chitinase homologue, demonstrating the difficulty when using sequence information to differentiate between endoxylanase inhibitors and (putative) chitinases in rice. Rice XIP inhibited different endoxylanases to a varying degree. In particular, it most strongly inhibited family 10 endoxylanases from A. niger and A. oryzae, while several family 11 enzymes from Bacillus subtilis, A. niger and Trichoderma sp. were not sensitive to inhibition. The above mentioned A. aculeatus endoxylanase was not inhibited either, although gel permeation chromatography revealed that it complexed rice XIP in a 1:1 molar stoichiometric ratio. PMID- 15895692 TI - Effects of nicotine and vitamin E on carbonic anhydrase activity in some rat tissues in vivo and in vitro. AB - Effects of nicotine, nicotine + vitamin E and nicotine + Hippophea rhamnoides L. extract (HRe-1) on muscle, heart, lungs, testicle, kidney, stomach, brain and liver carbonic anhydrase (CA; EC 4.2.1.1.) enzyme activities were investigated in vivo. Groups of rats were given nicotine (0.5 mg/kg/day, i.p.), nicotine + vitamin E (75 mg/kg/day, i.g.), nicotine + HRe-1 (250 mg/kg/day, i.g.) and a control group vehicle only. The results showed that nicotine inhibited the heart, lung, stomach and liver CA enzyme activities by approximately 80% (p < 0.001), approximately 94% (p < 0.001), approximately 47% (p < 0.001) and approximately 81% (p < 0.001) respectively, and activated muscle and kidney, but had no effects on the testicle and brain CA activities. Nicotine + vitamin E inhibited the heart and liver CA enzyme activities by approximately 50% (p < 0.001), and approximately 50% (p < 0.001), respectively, and nicotine + vitamin E activated the muscle CA activity. However, nicotine + vitamin E had no effect on lung, testicle, kidney, stomach and brain CA activities. Nicotine + HRe-1 inhibited the heart and stomach CA enzyme activities by approximately 51% (p < 0.001), and approximately 32% (p < 0.002), respectively, and activated the muscle and brain CA activities, but had no effects on the lung, testicle, kidney, and liver CA activities. In vitro CA inhibition results for similar experiments correlated well with the in vivo experimental results in lungs, testicles, kidney, stomach, brain and liver tissues. PMID- 15895693 TI - Monte Carlo simulations of the age structure of the human population. AB - In this mini review, we present assumptions and some results of Monte Carlo simulations based on the Penna model, which support the mutation accumulation theory of aging. This microscopic model has been exploited for the quantitative description of many biological phenomena connected with the population evolution. We show how the results of simulations could describe the changes of mortality trajectories of the human populations during the last 150 years, and how the method could be used for predicting the human age distribution in the future. The main assumption of the model is that genes are expressed chronologically one after another in the same order in all individuals during their life span. Different forces of selection pressure exerted on genes expressed at different periods of life generate characteristic gradient of defective genes accumulated in the germline cells. The genes expressed after the minimum reproduction age are under weaker selection pressure, and the fraction of defects among them is higher than among the genes expressed before the minimum reproduction age. This gradient of defects generates a gradient of mortality for the part of the population in the reproduction age following the exponential Gompertz law. The limitations of the model and some biological interpretations of its parameters are also discussed. PMID- 15895694 TI - Modeling disease elimination. AB - The effect of the elimination of mortality from heart disease and cancer was modelled mathematically to allow for the effect of other competing causes of death. The model allows for potential dependence between heart disease or cancer and other causes of death by using cupola functions, which analyse the individual risk itself and the dependence structure between causes of death by using correlation coefficients. As the strength of these risk associations is unknown, the study investigated both full positive and negative dependence and compared this with no dependence. Depending upon the degree and type of correlation assumed, positive or negative, the life expectancy at birth is increased by between 3 months and 6.5 years if cancer mortality was eliminated, and between 5 months and 7.5 years in the case of heart disease. In addition, estimates of these effects on life insurance premia can be made with the greatest reduction for women with the elimination of cancer mortality. These figures provide a range of improvements in life expectancy and the consequent effect on life insurance risk premium rates which elimination of either of these important diseases would produce. PMID- 15895695 TI - Estimation of future mortality rates and life expectancy in chronic medical conditions. AB - Estimates of old-age mortality are necessary for the construction of life tables and computation of life expectancy, and are essential in the growing area of life insurance for the elderly. Two common assumptions are that either the excess death rate (EDR) or the relative risk (RR) stays constant with increasing age. It is known, however, that for most medical conditions the former underestimates the risk and the latter overestimates it. A third popular method is that of rating up: a subject is said to be "rated up k years" if his future mortality rates are assumed to be those of a person in the general population who is k years older. It is shown here that this method generally leads to gross overestimates of old age mortality. We consider two less-commonly used models, log-linear declining relative risk (LDR) and constant proportional life expectancy (PLE), and compare them to the methods of constant EDR, constant RR and rating up. Although slightly more complicated to employ than the other methods, both LDR and PLE generally give better estimates of mortality and life expectancy. When mortality rates for chronic conditions are known within a certain age range, and estimates outside of the range are required, the LDR and PLE methods may be preferable to the more familiar methods of constant EDR, constant RR, or rating up. PMID- 15895696 TI - Estimating mortality rates: the role of proportional life expectancy. AB - The computation of long-term survival is usually based on adjustments to the conventional life table. Assessing the validity of different types of adjustments can be difficult, partly because of the need to allow for two age-related trends the decline in the average (normal) life expectancy, as well as in the new (abnormal) estimate. In this paper, we illustrate the value of routinely expressing each new estimate as a percentage of normal at each age. An additional finding has been that in some common disorders this proportional life expectancy (PLE) remains remarkably constant over many years. PMID- 15895697 TI - How to prepare a life expectancy report for an attorney in a tort case. AB - The purpose of this methodology article is to describe a suitable format for a legally acceptable report on the life expectancy of the principal in a tort case that is being advocated or defended by an attorney. Life insurance medical directors and underwriters are clearly skilled and experienced in mortality risk classification for life insurance. However, the judicial system is accustomed to measuring excess mortality only in terms of reduced life expectancy. The analyst preparing the report must convert the excess mortality into a figure for reduced life expectancy and compare this with the life expectancy of persons matched by age, sex and race in the latest Decennial US Life Tables. This process is different from the life insurance underwriting process. A life table projected to age 109 must be constructed as an essential part of the report, and the entire process must be presented clearly and convincingly. There are good reasons why the excess death rate (EDR) should be used as the index of excess mortality in constructing the life table, in preference to the mortality ratio (MR), which is used most of the time in life insurance risk classification. All of these considerations are discussed in this article, which is based on a sample of 40 cases handled by the author, a retired life insurance medical director. PMID- 15895698 TI - Coronary heart disease in the elderly--underwriting and pricing issues. AB - OBJECTIVE: The purpose of this analysis was to estimate the degree of excess mortality in elderly people with coronary heart disease. METHODS: Pertinent studies were found via an Internet search of English language articles in the National Library of Medicine. Excess mortality was estimated by the traditional calculations used in mortality methodology. RESULTS: Mortality ratios for coronary heart disease were very high at young ages and tapered with advancing age, reaching levels at ages 80 and older of 130% to 180%. Comparison with other mortality studies indicated that this pattern was typical for most impairments. Present value calculations showed that relative risk (mortality ratio) was lower at older ages, but absolute risk (mortality cost) was much higher. CONCLUSIONS: Mortality ratios for coronary heart disease and most common impairments are generally lower in elderly people. This has important underwriting and pricing implications. Disregarding what might be considered "low" mortality ratios in the elderly means that large additional risks are accepted with no additional premium. PMID- 15895699 TI - Poor R-wave progression. AB - Poor R-wave progression is a common ECG pattern, which is often inconclusively interpreted by medical directors. Although this ECG pattern is commonly attributed to anterior myocardial infarction, it may also be caused by left bundle branch block, Wolff-Parkinson-White syndrome, right and left ventricular hypertrophy as well as by faulty ECG recording technique. Failure to make a definitive interpretation of this pattern may result in a delay or loss of business. PMID- 15895701 TI - Artifacts in telephonic transmitted ECGs. PMID- 15895700 TI - Interesting electrocardiogram: the appearance of left bundle branch block during treadmill testing--revisited. PMID- 15895702 TI - Osteopetrosis. AB - This case study describes the fairly rare entity of osteopetrosis and points out how a seemingly insignificant "anemia" can, on thorough investigation, be found to be a very significant disease with a poor prognosis. PMID- 15895703 TI - Chronic cutaneous (discoid) lupus erythematosus. PMID- 15895704 TI - Atherosclerosis--an inflammatory process. PMID- 15895705 TI - Fabry disease and the heart: an overview of the natural history and the effect of enzyme replacement therapy. AB - Fabry disease is a genetic disorder caused by the deficiency of alpha galactosidase A, resulting in the lysosomal accumulation of glycosphingolipids. Fabry disease may result in cardiac, cerebral and renal complications. Cardiac abnormalities in patients with Fabry disease were first described in the 1960s. In the 1990s a form of Fabry disease confined to the heart was reported; however, this variant is extremely rare and a more appropriate concept is of cardiac predominance of the disease in some patients. Up to 60% of males with classic Fabry disease have cardiac abnormalities, including left ventricular hypertrophy, valvular dysfunction and conduction abnormalities. Recent data suggest that left ventricular mass and systolic function in patients with Fabry disease improve after 12 months of enzyme replacement therapy (ERT); however, many of the patients studied are relatively young and have mild cardiac abnormalities, suggesting that more research into the efficacy of ERT in older patients is necessary. CONCLUSION: Cardiac manifestations are common in patients with Fabry disease and are not confined to a 'cardiac variant' of the disease. PMID- 15895706 TI - The right ventricle in Fabry disease. AB - AIM: Left ventricular (LV) hypertrophy is a common feature in Fabry disease related progressive infiltrative hypertrophic cardiomyopathy and affects both men and women, but at different ages. To date, however, little is known about the role of right ventricular (RV) function in Fabry disease. Therefore, this study aimed to investigate the extent of RV involvement in patients with Fabry disease. METHODS: Echocardiographic examination of the right and left ventricle was carried out in 129 patients (80 women and 49 men) with Fabry disease. RESULTS: RV hypertrophy was present in 46 patients (35.7%). Of these patients, 13 showed signs of severely depressed right systolic function (tricuspid annulus movement < 10 mm and a prolonged RV pre-ejection period/pulmonary ejection time ratio) and six patients showed additional severe depression of parameters of diastolic function (pseudo-normal or restrictive RV filling pattems). Those patients with RV hypertrophy and severely compromised systolic and diastolic function had the highest LV masses (92 +/- 11.7 g/m(2.7)). CONCLUSION: RV involvement is common in Fabry disease and ultimately progresses to severe systolic and diastolic RV dysfunction. These findings might explain why patients with preserved LV function can develop clinical features such as reduced exercise capacity, organomegaly and lymphoedema. PMID- 15895707 TI - Measurement of renal function in patients with Fabry disease. AB - Appropriate measurement of the glomerular filtration rate (GFR) is important for the assessment of renal function. This paper reviews the methods used to assess GFR in clinical trials of enzyme replacement therapy (ERT) in patients with Fabry disease, which include inulin clearance, 24-hour creatinine clearance, chromium ethylene diamine tetraacetate (51Cr-EDTA) clearance and cystatin C concentrations. GFR has also been estimated using calculations based on creatinine clearance (the Cockcroft-Gault formula) and the Modification of Diet in Renal Disease (MDRD) equation. Analysis of the results of these studies shows that there are striking discrepancies between estimated and measured GFR. For example, the MDRD equation overestimates GFR in patients with Fabry disease who have normal renal function. In addition, cystatin C has been shown to be of limited use for measuring renal function during ERT, because it is influenced by other factors such as age, gender and weight. CONCLUSION: The use of exact methods, such as inulin clearance, 124I-iothalamate, 99mTc-DTPA, 51Cr-EDTA and iohexol, appears to be mandatory for a robust evaluation of the effects of ERT on GFR in patients with Fabry disease. PMID- 15895708 TI - Natural history of the cerebrovascular complications of Fabry disease. AB - Fabry disease is a rare, X-linked lysosomal storage disease caused by an inborn deficiency of alpha-galactosidase A, which results in progressive accumulation of globotriaosylceramide in a range of cells and tissues. Neurological symptoms of Fabry disease, including peripheral neuropathy and cerebrovascular events, are among the most significant clinical aspects. In this paper we present the natural history and mechanisms involved in the cerebrovascular complications of Fabry disease using data reported in FOS-- the Fabry Outcome Survey--and other registries and clinical studies. We discuss ways in which these manifestations can be modified by intervention, including both general measures for cerebrovascular disease and enzyme replacement therapy. CONCLUSION: Data from FOS have provided important insights into the natural history of the cerebrovascular complications of Fabry disease. Furthermore, the database has demonstrated that significant renal or cardiac disease often co-exists with cerebrovascular disease, and may predispose patients with Fabry disease to neurological disability and stroke. PMID- 15895709 TI - Vascular complications of Fabry disease: enzyme replacement and other therapies. AB - Fabry disease is an X-linked glycosphingolipid storage disorder resulting from deficiency of alpha-galactosidase A. Storage of globotriaosylceramide ultimately results in multiorgan pathology, including cerebrovascular, cardiovascular and renal disease. Vascular involvement is evident throughout the body but the mechanisms by which storage on a cellular level leads to end-organ pathology are unknown. Here the evidence for abnormal blood flow, vessel architecture and endothelial function will be reviewed and possible models of vascular pathology discussed. The effects of reversal of storage within vessels by enzyme replacement therapy (ERT) and the possibilities for intervention with additional agents will be considered. CONCLUSION: The pathology of Fabry disease has an important vascular component, although the underlying pathophysiology is unclear. Preliminary evidence suggests that ERT may have beneficial effects on the vascular component of this multisystem disease. PMID- 15895710 TI - Biomarkers in lysosomal storage diseases: a review. AB - A biomarker is generally an analyte that indicates the presence or extent of a biological process, which is itself directly linked to the clinical manifestations and outcome of a particular disease. An ideal biomarker provides indirect but ongoing and specific determinations of disease activity. These characteristics emphasize the value of surrogate biomarkers for non-invasive and detailed monitoring to demonstrate the efficacy of orphan drugs in clinical trials. The emergence of novel laboratory methods has facilitated the search for biomarkers in lysosomal storage diseases (LSDs), by allowing the systematic identification of molecules whose expression is altered as a result of the primary storage pathology. In Gaucher disease, for example, a chemokine, CCL18, has been identified as a biomarker for clinical development that reflects disease severity and response to treatment. CONCLUSION: New methods for the identification of novel biomarkers have the potential to provide mechanistic insights into the molecular pathogenesis of LSDs, including Fabry disease and Gaucher disease. PMID- 15895711 TI - Identification and use of biomarkers in Gaucher disease and other lysosomal storage diseases. AB - The value of biomarkers in the clinical management of lysosomal storage diseases is best illustrated by the present use of plasma chitotriosidase levels in the diagnosis and monitoring of Gaucher disease. The enzyme chitotriosidase is specifically produced and secreted by the pathological storage macrophages (Gaucher cells). Plasma chitotriosidase levels are elevated on average 1000-fold in symptomatic patients with Gaucher disease and reflect the body burden on storage cells. Changes in plasma chitotriosidase reflect changes in clinical symptoms. Monitoring of plasma chitotriosidase levels is nowadays commonly used in decision making regarding initiation and optimization of costly therapeutic interventions (enzyme replacement therapy or substrate reduction therapy). A novel substrate has been developed that further facilitates the measurement of chitotriosidase in plasma samples. Moreover, an alternative Gaucher-cell marker, CCL18, has been very recently identified and can also be employed to monitor the disease, particularly in those patients lacking chitotriosidase due to a genetic mutation. There is a need for comparable surrogate markers for other lysosomal storage diseases and the search for such molecules is an area of intense investigation. CONCLUSION: The use of biomarkers can provide valuable insight into the molecular pathogenesis of LSDs, such as Gaucher disease and Fabry disease. PMID- 15895712 TI - Clinical evaluation of biomarkers in Gaucher disease. AB - Novel or candidate biomarkers require thorough evaluation to establish their utility in a clinical setting. This paper describes an evaluation of several established enzyme markers of Gaucher disease and a newly-described chemokine, pulmonary and activation-regulated chemokine (PARC). The ability of the biomarkers to rank patients with Gaucher disease in order of disease severity and organ bulk, and to reflect changes in key clinical parameters in response to enzyme replacement therapy were evaluated. PARC concentrations were found to be reliably correlated with visceral disease and with key clinical responses to enzyme replacement in an unbiased manner. Unlike chitotriosidase and serum angiotensin-converting enzyme activity, genetic variation in serum PARC did not appear to influence its utility as a biomarker. CONCLUSION: For each new candidate biomarker of lysosomal storage diseases, a similar clinical evaluation will be required, though the approach will need to be modified according to the clinical features and natural history of each disorder. PMID- 15895713 TI - Is globotriaosylceramide a useful biomarker in Fabry disease? AB - AIM: The aim of this study was to determine whether globotriaosylceramide (Gb3) is a useful biomarker in Fabry disease. METHODS: The levels of Gb3 were measured in plasma and urine by tandem mass spectrometry in untreated hemizygotes and heterozygotes with Fabry disease and in healthy controls, and the levels were monitored in patients on treatment with enzyme replacement therapy (ERT). RESULTS: Hemizygotes with classic Fabry disease showed elevated levels of Gb3 in both plasma and urine and could readily be distinguished from normal controls. Male patients with the N215S mutation had normal levels in their plasma but 50% had marginally elevated levels in their urine. Thirty-three percent of proven heterozygotes had elevated Gb3 concentrations in plasma but 97% of those without the N215S mutation (36/37) had an elevated level in urine. The four heterozygotes with the N215S mutation had normal Gb3 levels in urine. The level of Gb3 in plasma initially fell following the start of ERT in all patients who had an elevated level before treatment. However, in a few patients the level subsequently rose. Similar results were found for the levels of Gb3 in urine. CONCLUSION: Gb3 is not an ideal marker of Fabry disease or the response to treatment in all patients. Plasma and urine levels of Gb3 cannot be used as a marker of Fabry disease in patients with the N215S mutation and many heterozygotes do not have elevated Gb3 levels in plasma. The urine concentration is more informative in heterozygotes and can be used as a measure of the response to therapy. The fall in Gb3 levels in patients receiving ERT was not sustained in some patients, despite a clinical improvement. Additionally, Gb3 cannot be used to monitor the response to treatment in patients who initially have normal plasma and urine concentrations of this glycolipid. PMID- 15895714 TI - Enzyme replacement therapy in mucopolysaccharidosis type I. AB - Mucopolysaccharidosis (MPS) type I is a lysosomal storage disorder caused by deficiency of the enzyme alpha-L-iduronidase (IDUA), which presents with a wide spectrum of phenotypes. Recently, enzyme replacement therapy (ERT) became available for patients with MPS I and has been demonstrated to be safe and effective in patients with the milder Hurler-Scheie and Scheie phenotypes. Treatment for 26 weeks with recombinant human IDUA (laronidase) has been shown to significantly increase the percentage of predicted normal forced vital capacity and the distance walked in the 6-minute walk test. There was also a clear reduction in the volume of the liver and the levels of urinary glycosaminoglycan excretion. The drug was generally well tolerated. There were no drug-related severe adverse events, and although the majority of patients developed IgG antibodies, these declined by the end of the study. CONCLUSION: ERT seems to be a very promising new therapeutic regimen for patients with MPS I, especially for those with the less severe variants. However, as laronidase does not cross the blood-brain barrier it will probably not influence the central nervous manifestations in the most severely affected patients with the Hurler phenotype, although it may improve general lung and heart function, making bone marrow transplantation easier to tolerate. PMID- 15895715 TI - Pharmacokinetic profile of recombinant human N-acetylgalactosamine 4-sulphatase enzyme replacement therapy in patients with mucopolysaccharidosis VI (Maroteaux Lamy syndrome): a phase I/II study. AB - AIM: Mucopolysaccharidosis VI (Maroteaux-Lamy syndrome) is a lysosomal storage disease caused by a deficiency of the enzyme-N-acetylgalactosamine 4-sulphatase (ASB). Enzyme replacement therapy with recombinant human ASB (rhASB) has been studied in a randomized, double-blind, two-dose (0.2 and 1.0 mg/kg/week) phase I/II study (n = 7) followed by an open-label single dose (1.0 mg/kg/week) extension study. We report the pharmacokinetic profile of rhASB and the impact of antibody development. METHODS: Pharmacokinetic analysis was performed at weeks 1, 2, 12, 24, 83, 84 and 96. Infusions were administered over 4 hours using a ramp up protocol. Plasma ASB and rhASB antibody concentrations and urine glycosaminoglycan (GAG) concentrations were determined. RESULTS: The area under the plasma concentration-time curve (AUC(0-t)) for the high-dose group increased from week 1 to week 2, but remained unchanged at weeks 12 and 24. A large difference in mean AUC(0-t) was observed between the low- and high-dose groups. Pharmacokinetic results at weeks 83, 84 and 96 were similar to those at week 24. Six patients developed antibodies to rhASB. One patient developed high antibody levels in combination with a high ASB concentration, while a second patient also developed high antibody levels with undetectable ASB concentrations. Antibodies from the second patient blocked detection of ASB. By week 72, antibody levels had decreased in all patients. The high-dose rhASB produced a more rapid and greater percentage reduction in urinary GAG concentrations than the lower dose (70% versus 55% at 24 weeks). Antibody levels did not appear to influence urinary GAG concentrations. CONCLUSION: Pharmacokinetic parameters appear to be independent of the duration of treatment and are not linear between the 0.2 and 1.0 mg/kg/week doses. Antibodies to rhASB develop in most patients, but their concentration decreases over time. Antibody formation may influence pharmacokinetic parameters during the early phases of treatment, although it appears to have limited impact on biochemical efficacy. PMID- 15895716 TI - Substrate reduction therapy for lysosomal storage diseases. AB - The treatment of disordered lipoprotein metabolism with the statin class of drugs is one of the most striking successes in the field of applied medical science: here the use of selective inhibitors of the first committed step of cholesterol biosynthesis, in a complex and highly regulated pathway, leads to improved outcome from a common lipid storage disease that is a blight on whole populations -atherosclerosis. By the same token, substrate reduction is an emerging therapeutic strategy for the arcane field of the lysosomal storage diseases (LSDs). Reduced biosynthesis of glucosylceramide is postulated to allow correction of the imbalance between formation and breakdown of glycosphingolipids; the therapeutic effect of substrate reduction depends upon the presence of residual hydrolytic activity towards those accumulated glycosphingolipid substrates derived from glucosylceramide. First pioneered in the laboratory by Norman Radin, this approach has now been introduced into the clinic: based on the ability to inhibit uridine diphosphate glucosylceramide transferase, the semi-selective iminosugar, N-butyldeoxynojirimycin, is licensed for the treatment of type 1 Gaucher disease. CONCLUSION: Inhibition of substrate formation has wide application in the treatment of LSDs. Decreased glucosylceramide biosynthesis has therapeutic potential in glycosphingolipidoses other than Gaucher disease, and offers promise in several neurodegenerative storage disorders that are currently beyond the reach of other procedures. The results of ongoing clinical trials of miglustat in type 3 Gaucher disease, Niemann-Pick disease type C and GM2 gangliosidosis are eagerly awaited. PMID- 15895717 TI - What can cell biology tell us about heterogeneity in lysosomal storage diseases? AB - Lysosomal storage diseases are clinically heterogeneous with respect to their age of onset, progression of symptoms and the particular organs involved. Varying levels of residual enzyme activity, associated with different defective alleles that cause the respective diseases, are responsible in part for this clinical heterogeneity. In general, the higher the residual enzyme activity, the milder the phenotype. Enzyme activity in severe forms of disease is frequently zero, and in mild forms usually does not exceed approximately 5%. However, the correlation is not so strict as to allow prediction of the phenotype of individual patients. The molecular basis of the different levels of enzyme activity can only be revealed by biochemical investigations of the defective lysosomal proteins. Null alleles may be due to splice-site mutations or deletions. In the case of missense mutations, enzymes frequently fold incorrectly and are retained in the endoplasmic reticulum and subsequently degraded. As these enzymes do not reach the lysosome, they do not provide any functional residual activity. Residual enzyme activity is only observed in cases where the defective enzyme reaches the lysosome and has retained enzymatic activity. Patients carrying the same mutant alleles still show considerable phenotypic variability due to modifying genes and epigenetic factors. None of these has so far been elucidated. However, there are some indications that differences in splicing-factor machinery may influence the phenotypic expression of splice-site mutations and that hormonal modulation of secondary microglial activation in lipidosis may also influence the disease course. CONCLUSION: Phenotypic variability is a frequent phenomenon in lysosomal storage diseases. Residual enzyme activity has been identified as one of the factors influencing the clinical outcome of disease; however, it is obvious that other genetic and epigenetic factors also affect phenotypic variability, particularly in patients with late onset disease. PMID- 15895718 TI - Genotype and phenotype in Fabry disease: analysis of the Fabry Outcome Survey. AB - AIM: Mutations of the gene (GLA) encoding alpha-galactosidase A are implicated in Fabry disease, a progressive, X-chromosomal inherited lysosomal storage disorder. FOS--the Fabry Outcome Survey - was established as a long-term surveillance study to describe the natural course of Fabry disease and its response to enzyme replacement therapy in a large cohort of European patients. Clinical phenotype, age of onset and course of Fabry disease are very variable, even within the same family, which makes it difficult to define a genotype-phenotype relationship by analysing individual patients. The FOS database contains detailed medical information on a large cohort of patients and thus has the potential to provide important information to address this question. METHODS: At the time of analysis, information on 545 patients belonging to 157 families from nine European countries had been entered into the FOS database. A GLA mutation has been reported in 365 individuals (65% and 68% of all males and females, respectively) in FOS. These data were used to analyse the relationship between genotype and phenotype in Fabry disease. RESULTS: A highly significant positive correlation was found between the age at entry into FOS and the FOS Severity Index in male patients with GLA missense mutations (p < 0.001) as well as in those carrying other types of mutations (p < 0.001). A positive correlation was also found between the age at entry into FOS and the number of affected organs in male patients with missense mutations, irrespective of whether the change in the amino acid side chain predicted in the alpha-galactosidase A protein was classified as a conservative or non-conservative change. CONCLUSION: The data presented here suggest that there is a correlation between genotype and clinical severity. PMID- 15895719 TI - Persistence of Escherichia coli and Salmonella in surface soil following application of liquid hog manure for production of pickling cucumbers. AB - Liquid hog manure is routinely applied to farm land as a crop fertilizer. However, this practice raises food safety concerns, especially when manure is used on fruit and vegetable crops. The objectives of this project were to evaluate the persistence of Escherichia coli and Salmonella in surface soil after application of liquid hog manure to fields where pickling cucumbers were grown and to verify the microbiological quality of harvested cucumbers. Mineral fertilizers were replaced by liquid hog manure at various ratios in the production of pickling cucumbers in a 3-year field study. The experimental design was a randomized complete block comprising four replicates in sandy loam (years 1, 2, and 3) and loamy sand (year 3). Soil samples were taken at a depth of 20 cm every 2 weeks after June application of organic and inorganic fertilizers. Vegetable samples were also taken at harvest time. Liquid hog manure, soil, and vegetable (washed and unwashed) samples were analyzed for the presence of Salmonella and E. coli. An exponential decrease of E. coli populations was observed in surface soil after the application of manure. The estimated average time required to reach undetectable concentrations of E. coli in sandy loam varied from 56 to 70 days, whereas the absence of E. coli was estimated at 77 days in loamy sand. The maximal Salmonella persistence in soil was 54 days. E. coli and Salmonella were not detected in any vegetable samples. PMID- 15895720 TI - Biofilm formation, cellulose production, and curli biosynthesis by Salmonella originating from produce, animal, and clinical sources. AB - The ability of 71 strains of Salmonella enterica originating from produce, meat, or clinical sources to form biofilms was investigated. A crystal violet binding assay demonstrated no significant differences in biofilm formation by isolates from any source when tested in any of the following three media: Luria-Bertani broth supplemented with 2% glucose, tryptic soy broth (TSB), or 1/20th-strength TSB. Incubation was overnight at 30 degrees C under static conditions. Curli production and cellulose production were monitored by assessing morphotypes on Luria-Bertani agar without salt containing Congo red and by assessing fluorescence on Luria-Bertani agar containing calcofluor, respectively. One hundred percent of the clinical isolates exhibited curli biosynthesis, and 73% demonstrated cellulose production. All meat-related isolates formed curli, and 84% produced cellulose. A total of 80% of produce-related isolates produced curli, but only 52% produced cellulose. Crystal violet binding was not statistically different between isolates representing the three morphotypes when grown in TSB; however, significant differences were observed when strains were cultured in the two other media tested. These data demonstrate that the ability to form biofilms is not dependent on the source of the test isolate and suggest a relationship between crystal violet binding and morphotype, with curli- and cellulose-deficient isolates being least effective in biofilm formation. PMID- 15895721 TI - Application of microbial risk assessment to the development of standards for enteric pathogens in water used to irrigate fresh produce. AB - Microbial contamination of the surfaces of cantaloupe, iceberg lettuce, and bell peppers via contact with irrigation water was investigated to aid in the development of irrigation water quality standards for enteric bacteria and viruses. Furrow and subsurface drip irrigation methods were evaluated with the use of nonpathogenic surrogates, coliphage PRD1, and Escherichia coli ATCC 25922. The concentrations of hepatitis A virus (HAV) and Salmonella in irrigation water necessary to achieve a 1:10,000 annual risk of infection, the acceptable level of risk used for drinking water by the U.S. Environmental Protection Agency, were calculated with a quantitative microbial risk assessment approach. These calculations were based on the transfer of the selected nonpathogenic surrogates to fresh produce via irrigation water, as well as previously determined preharvest inactivation rates of pathogenic microorganisms on the surfaces of fresh produce. The risk of infection was found to be variable depending on type of crop, irrigation method, and days between last irrigation event and harvest. The worst-case scenario, in which produce is harvested and consumed the day after the last irrigation event and maximum exposure is assumed, indicated that concentrations of 2.5 CFU/100 ml of Salmonella and 2.5 x 10(-5) most probable number per 100 ml of HAV in irrigation water would result in an annual risk of 1:10,000 when the crop was consumed. If 14 days elapsed before harvest, allowing for die-off of the pathogens, the concentrations were increased to 5.7 x 10(3) Salmonella per 100 ml and 9.9 x 10(-3) HAV per 100 ml. PMID- 15895722 TI - Increase in activity of essential oil components carvacrol and thymol against Escherichia coli O157:H7 by addition of food stabilizers. AB - The major components of oregano and thyme essential oils that had previously been shown to inhibit Escherichia coli O157:H7 were determined by high-performance liquid chromatography with UV detection and liquid chromatographic tandem mass spectrometry. The MICs and MBCs of carvacrol, thymol, p-cymene, and gamma terpinene against a strain of E. coli O157: H7 phage type 34 isolated from bovine feces were determined by microdilution assay. The constituents were then tested in checkerboard assays to detect possible interactions. Carvacrol and thymol displayed bacteriostatic and bactericidal properties with MICs of 1.2 mmol/liter and were additive in combination. p-Cymene and gamma-terpinene displayed no measurable antibacterial activity up to 50 mmol/liter, and neither influenced the activity of carvacrol or thymol. Growth curves in the presence of nonlethal concentrations of carvacrol with the addition of agar (0.05%, wt/vol) or carrageenan (0.125%, wt/vol) as stabilizer were produced by optical density measurement. The stabilizers agar and carrageenan both significantly improved the effectiveness of carvacrol in broth, possibly because of a delay in the separation of the hydrophobic substrate from the aqueous phase of the medium. When carvacrol was dissolved in ethanol before addition to broth, stabilizers were not needed. Carvacrol and thymol, particularly when used in combination with a stabilizer or in an ethanol solution, may be effective in reducing the number or preventing growth of E. coli O157:H7 in liquid foods. PMID- 15895723 TI - Escherichia coli O157:H7, Campylobacter jejuni, and Salmonella Prevalence in cull dairy cows marketed in northeastern Ohio. AB - Preharvest management factors are predicted to impact the prevalence of foodborne pathogens in cattle sent to slaughter. We simultaneously examined the prevalence and antibiotic resistance patterns of Campylobacter jejuni, Salmonella, and Escherichia coli O157:H7 isolated from cull dairy cattle at two livestock auctions in northeastern Ohio. Between April and September 2002, a total of 1,026 fecal samples were collected. C. jejuni was isolated from 48 of 686 (7%) fecal samples, Salmonella was isolated from 39 of 585 (6.7%) samples, and E. coli O157:H7 was isolated from 21 of 1,026 (2.1%) samples. Of the 585 samples tested for all three pathogens, at least one pathogen was identified in 86 of 585 (15%) samples. One sample was positive for both E. coli O157:H7 and C. jejuni, and five samples yielded both C. jejuni and Salmonella. Size of herd of origin could be traced for 75 to 85% of samples collected. Salmonella was isolated at higher frequencies from herds larger than 60 cattle than from smaller herds (9.0 versus 3.5%, P = 0.02). In contrast, size of herd of origin did not significantly affect the E. coli O157:H7 and C. jejuni prevalence. Approximately 90% of Salmonella and E. coli O157:H7 isolates were pansensitive to a panel of 16 antibiotics. Thirty six percent of C. jejuni isolates were resistant to tetracycline. In this study, antibiotic resistance among the foodborne pathogens isolated from cull diary cattle was rare. Although size of dairy herd of origin was positively associated with Salmonella prevalence, herd size was not strongly associated with E. coli O157:H7 and C. jejuni prevalence in market dairy cattle. These results can be used to assess the food safety risks associated with the slaughter of cull dairy cattle. PMID- 15895724 TI - Identification of factors involved in recovery of heat-injured Salmonella Enteritidis. AB - Proteins and genes involved in the recovery of heat-injured Salmonella Enteritidis were investigated. Salmonella Enteritidis cells cultured overnight in tryptic soy broth (TSB; nonselective medium) were suspended in citric acid disodium hydrogen phosphate buffer (pH 6). After heat treatment at 55 degrees C for 15 min, the culturable counts measured by tryptic soy agar (TSA; nonselective medium) decreased from 10(8) to 10(7) CFU/ml. On the other hand, culturable counts measured by desoxycholate-hydrogen sulfite-lactose (DHL) agar (selective medium) were decreased from 10(8) to 10(4) CFU/ml by the same treatment. The results suggest that 99.9% of Salmonella Enteritidis detected on TSA were injured but recoverable. When injured Salmonella Enteritidis was incubated in TSB, the culturable count measured by TSA did not increase for 2 h, whereas that by DHL agar increased after incubation for 30 min. After incubation for 2 h, the culturable count measured by DHL agar reached a similar level with that by TSA, indicating that Salmonella Enteritidis had recovered. The two-dimensional polyacrylamide gel electrophoresis analysis revealed that elongation factor G (FusA) and pyruvate kinase (PykF) specifically increased in the cells just after heat treatment and in the recovery cells. The levels of transcription of 86 stress-inducible genes were also investigated by reverse transcription PCR. Nineteen heat-inducible (clpB, clpX, degP, dnaJ, fkpA, ftsJ, gapA, hflB, hslJ, hslU, hslV, htpG, htrA, lon, mopA, mopB, mreB, rpoE, and ppiD), and 12 oxidative stress and DNA damage-inducible (ahpC, ahpF, fldB, fur, grxA, dinF, katG, mutM, recA, soxR, trxC, and zwf) genes were transcribed extensively during recovery in TSB. The results obtained in this study will be used to develop the media or culture conditions that will promote recovery for the detection of food poisoning bacteria, including injured cells from food products. PMID- 15895725 TI - Thermal resistance of heat-, cold-, and starvation-injured Salmonella in irradiated comminuted Turkey. AB - To investigate the effects of sublethal stress on Salmonella thermal inactivation kinetics, an eight-strain Salmonella cocktail was subjected to heat shock (30 min at 54 degrees C), cold shock (2 h at 4 degrees C), and starvation stress (10 days in phosphate buffer at 4 degrees C), harvested by centrifugation, and inoculated into irradiated comminuted turkey. Immediately after stressing, the Salmonella cocktails contained 89.1% heat-injured, 44.7% cold-injured, and 67.7% starvation injured cells, as determined by plating on selective and nonselective media. D60 degrees C-values for the heat-shocked cocktail (0.64 min on Trypticase soy agar containing 0.6% yeast extract [TSAYE], 0.35 min on xylose lysine desoxycholate [XLD] agar) were higher (P < 0.05) than those for the unshocked control (0.41 min on TSAYE, 0.17 min on XLD), whereas D60 degrees -values for the cold-shocked cocktail (0.38 min on TSAYE, 0.17 min on XLD) were not significantly different from those for the control. Starved cells had the same D60 degrees C-value on TSAYE as did the unshocked cocktail, but the D60 degrees C-value on XLD was significantly lower (0.14 min). Although starvation and cold shock were not thermally protective, heat shock increased thermal resistance, indicating that product history and the physiological state of the Salmonella cells should be considered when developing and validating thermal processes. D60 degrees C-values observed on selective media were significantly lower than those observed on nonselective media for all stress treatments and for the control. Therefore, nonselective culture media should be used to assess the response of microorganisms to a thermal challenge when developing performance standards for lethality. PMID- 15895726 TI - Toward validation of process criteria for high-pressure processing of orange juice with predictive models. AB - Mathematical models were developed to predict time to inactivation (TTI) by high pressure processing of Salmonella in Australian Valencia orange juice (pH 4.3) and navel orange juice (pH 3.7) as a function of pressure magnitude (300 to 600 MPa) and inoculum level (3 to 7 log CFU/ml). For each model, the TTI was found to increase with increasing inoculum level and decrease with increasing pressure magnitude. The U.S. Food and Drug Administration Juice Hazard Analysis and Critical Control Point Regulation requires fruit juice processors to include control measures that produce a 5-log reduction of the pertinent microorganism of public health significance in the juice. To achieve a 5-log reduction of Salmonella in navel orange juice at 20 degrees C, the models predicted hold times of 198, 19, and 5 s at 300, 450, and 600 MPa, respectively. In Valencia orange juice at 20 degrees C, a 5-log reduction of Salmonella was achieved in 369, 25, and 5 s at 300, 450, and 600 MPa, respectively. At pressures below 400 MPa, Salmonella was more sensitive to pressure in the more acidic conditions of the navel orange juice and TTIs were shorter. At higher pressures, little difference in the predicted TTI was observed. Refrigerated storage (4 degrees C) of inoculated navel orange juice treated at selected pressure/time/inoculum combinations showed that under conditions in which viable Salmonella was recovered immediately after high-pressure processing, pressure-treated Salmonella was susceptible to the acidic environment of orange juice or to chill storage temperature. These TTI models can assist fruit juice processors in selecting processing criteria to achieve an appropriate performance criterion with regard to the reduction of Salmonella in orange juice, while allowing for processing flexibility and optimization of high-pressure juice processing. PMID- 15895727 TI - Contribution of enterococci to the spread of antibiotic resistance in the production chain of swine meat commodities. AB - Thirty-six samples, including fecal specimens, dry feedstuffs, raw and processed pork meat products, and dry fermented sausages, were collected from two production chains of swine meat commodities and analyzed for the presence of 11 antibiotic resistance (AR) genes. Specific PCR assays carried out on DNA extracted directly from the samples revealed a high incidence of the genes tet(K) (80.5%), ermB (66.7%), and tet(M) (66.7%). Feces and feedstuffs gave the largest number of positive amplifications. To elucidate the contribution of enterococci to the occurrence and spread of AR, 146 resistant enterococci were isolated, and their identity, genetic fingerprints, and AR gene profiles were determined by means of molecular techniques. Enterococcus faecalis and Enterococcus faecium were the predominant isolated species (43.8 and 38.4%, respectively); Other Enterococcus species identified were E. durans (8.9%), E. hirae (2.7%), E. gallinarum (2.1%), E. mundtii (2.1%), and E. casseliflavus (2.1%). A number of isolates displayed a complex AR gene profile comprising up to four different resistance determinants. The genes tet(M) and ermB were highly diffused, being present in 86.9 and 84.9%, respectively, of the isolates. The application of amplified fragment length polymorphism fingerprinting was particularly valuable to monitor the resistant enterococcal isolates along the production chain and to individuate steps in which contamination might occur. In fact, isolates of E. faecalis and E. faecium showing the same amplified fragment length polymorphism profile and AR gene pattern were detected in samples taken at different steps of the food chain suggesting three cases of bacterial clonal spread. PMID- 15895728 TI - Detection of viable Mycobacterium avium subsp. paratuberculosis in retail pasteurized whole milk by two culture methods and PCR. AB - Cattle with Johne's disease can shed live Mycobacterium avium subsp. paratuberculosis (MAP) in their milk, and MAP can survive under simulated commercial pasteurization conditions. In several studies conducted in the United Kingdom and Canada, MAP DNA has been detected in retail pasteurized milk samples; however, in one study in the United Kingdom viable MAP was identified in commercially pasteurized milk. A double-blind study involving two laboratories was undertaken to evaluate retail pasteurized whole milk in the United States. Marshfield Clinic Laboratories used solid culture medium (Herrold's egg yolk agar slants with mycobactin J and amphotericin B, nalidixic acid, and vancomycin), and TREK Diagnostic Systems, Research and Development used liquid culture medium (ESP culture system). Cultures at both laboratories were eonfirmed by PCR. A total of 702 pints of retail whole milk were purchased in three of the top five milk producing states (233 from California, 234 from Minnesota, and 235 from Wisconsin) over a 12-month period and were tested for the presence of viable MAP. The criteria used for identifying samples as positive for viable MAP were similar to those followed by most laboratories (positive culture with PCR confirmation). The combined data from the two laboratories revealed the presence of viable MAP in 2.8% of the retail whole milk pints tested. Although the number of samples containing viable MAP was similar among states (P > 0.05), there was a seasonal effect on the presence of viable MAP in retail milk (P = 0.05). More MAP-positive samples were identified during the third quarter of the year (July through September). Of the 22 brands of retail milk tested, 12 (55%) yielded at least one sample positive for viable MAP. PMID- 15895729 TI - Comprehensive survey of pasteurized fluid milk produced in the United States reveals a low prevalence of Listeria monocytogenes. AB - A comprehensive survey was undertaken to generate contemporary data on the prevalence of Listeria monocytogenes in pasteurized fluid milk produced in the United States. Samples (5,519) near the sell-by expiration date were purchased at retail outlets over a 5-week period and analyzed for presence of L. monocytogenes. Products consisted of whole milk, nonfat milk, and chocolate milk packaged in gallon, half gallon, quart, pint, and half-pint containers. Samples were collected from both large and small retail stores in urban and suburban locations in four FoodNet cities (Baltimore, Md., Atlanta, Ga., St. Paul/ Minneapolis, Minn., and San Francisco, Calif.). Samples were prescreened for L. monocytogenes by the AOAC-approved rapid Vitek immunodiagnostic assay system, enzyme-linked fluorescent assay method. Positive prescreening samples were cultured according to the Bacteriological Analytical Manual, enumerated for L. monocytogenes with a nine-tube most-probable-number (MPN) procedure, and confirmed by biochemical characterization. The frequency of isolation of L. monocytogenes in these products was 0% (0 of 1,897) in whole milk, 0.05% (1 of 1,846) in nonfat milk, 0% (0 of 1,669) in chocolate milk, and 0% (0 of 107) in other (reduced fat and low fat) milk samples. Overall, L. monocytogenes was confirmed in only 0.018% of pasteurized milk samples (1 of 5,519). Enumeration of the single confirmed positive nonfat milk sample revealed low-level contamination (<0.3 MPN/g), even when sampled 5 days past the expiration of the sell-by date. The results confirm the low frequency of contamination of pasteurized fluid milk products by L. monocytogenes for products sold in the United States and reaffirm the reduction of contamination frequency of fluid milk by L. monocytogenes when compared with earlier estimates from the U.S. Food and Drug Administration Dairy Safety Initiatives Program. PMID- 15895730 TI - Distribution of Listeria monocytogenes subtypes within a poultry further processing plant. AB - Samples from environmental sites and raw product in a chicken further processing plant were collected every 6 weeks for 12 months. Each sample site was examined before and after a complete production shift. All samples were examined for the presence of Listeria monocytogenes, which was detected in floor drains on the raw product side of the plant preoperation and in drains on both raw and cooked sides following 8 h of processing operation. L. monocytogenes also was detected in raw product and once in fully cooked product but never on cooked product contact surfaces. One hundred sixty-one isolates were collected from 75 positive samples. All isolates were subtyped using a sequence-based method, and 14 unique subtypes were detected through the course of the study. Four of these types were found repeatedly and appeared to be resident in the plant. Three of the four resident strains were detected on raw product at some point during the year-long study, suggesting that raw product may be one source of L. monocytogenes in the processing plant environment. These data highlight the need for research to investigate why some types of L. monocytogenes persist in a processing plant environment but others do not. PMID- 15895731 TI - Efficacy of electrolyzed water in inactivating Salmonella enteritidis and Listeria monocytogenes on shell eggs. AB - The efficacy of acidic electrolyzed (EO) water produced at three levels of total available chlorine (16, 41, and 77 mg/ liter) and chlorinated water with 45 and 200 mg/liter of residual chlorine was investigated for inactivating Salmonella Enteritidis and Listeria monocytogenes on shell eggs. An increasing reduction in Listeria population was observed with increasing chlorine concentration from 16 to 77 mg/liter and treatment time from 1 to 5 min, resulting in a maximal reduction of 3.70 log CFU per shell egg compared with a deionized water wash for 5 min. There was no significant difference in antibacterial activities against Salmonella and Listeria at the same treatment time between 45 mg/liter of chlorinated water and 14-A acidic EO water treatment (P > or = 0.05). Chlorinated water (200 mg/liter) wash for 3 and 5 min was the most effective treatment; it reduced mean populations of Listeria and Salmonella on inoculated eggs by 4.89 and 3.83 log CFU/shell egg, respectively. However, reductions (log CFU/shell egg) of Listeria (4.39) and Salmonella (3.66) by 1-min alkaline EO water treatment followed by another 1 min of 14-A acidic EO water (41 mg/liter chlorine) treatment had a similar reduction to the 1-min 200 mg/liter chlorinated water treatment for Listeria (4.01) and Salmonella (3.81). This study demonstrated that a combination of alkaline and acidic EO water wash is equivalent to 200 mg/liter of chlorinated water wash for reducing populations of Salmonella Enteritidis and L. monocytogenes on shell eggs. PMID- 15895732 TI - Postprocessing antimicrobial treatments to control Listeria monocytogenes in commercial vacuum-packaged bologna and ham stored at 10 degrees C. AB - The antilisterial effect of chemical dipping solutions on commercial bologna and ham slices, inoculated (3 to 4 log CFU/ cm2) after processing, was evaluated during storage in vacuum packages at 10 degrees C. Samples were inoculated with a 10-strain composite of Listeria monocytogenes and subsequently immersed (25+/-2 degrees C) for 2 min in 2.5% acetic acid (AA), 2.5% lactic acid (LA), 5% potassium benzoate (PB), or 0.5% Nisaplin (commercial form of nisin, equivalent to 5,000 IU/ml of nisin) solutions, either singly or sequentially (Nisaplin plus AA, Nisaplin plus LA, or Nisaplin plus PB), and then vacuum packaged and stored at 10 degrees C for 48 days. In addition to microbiological analysis, sensory evaluations were performed on uninoculated samples treated with AA, LA, or PB. Initial reductions (day 0) of the pathogen, compared with the controls, on bologna and ham samples treated with AA, LA, or PB ranged from 0.4 to 0.7 log CFU/cm2. Higher (P < 0.05) initial reductions (2.4 to 2.9 log CFU/cm2) were obtained for samples treated with Nisaplin alone and when followed by AA, LA, or PB. L. monocytogenes populations on control bologna and ham samples increased from 3.4 log CFU/cm2 (day 0) to 7.4 and 7.8 log CFU/ cm2, respectively, in 8 days at 10 degrees C. Listericidal effects were observed for all treatments tested, except for Nisaplin applied on its own, during storage at 10 degrees C. The sequential treatment of Nisaplin plus LA reduced L. monocytogenes to undetectable levels in both products at the end of storage. The sequential treatments were also found to inhibit growth of spoilage microorganisms. Sensory evaluations indicated that dipping (2 min) of ham samples in AA (2.5%), LA (2.5%), or PB (5%) led to lower sensory scores. However, since results of this study indicated that these treatments caused extensive listericidal effects, there is possibly a potential to reduce the levels of chemicals applied and still achieve adequate antilisterial activity without major negative effects on product quality. PMID- 15895733 TI - Prewashing with acidified sodium chlorite reduces pathogenic bacteria in lightly fermented Chinese cabbage. AB - Efficacy of prewashing with acidified sodium chlorite (ASC) for the sanitation of lightly fermented Chinese cabbage was evaluated. The population of the natural microflora on the cabbage leaves was reduced about 2.0 log CFU/g just after washing with ASC, a significant reduction compared with the control distilled water wash (P < or = 0.05). In the control experiment, viable aerobic bacteria increased gradually when incubated at 10 degrees C; however, ASC-washed cabbage maintained a lower microbial concentration. The treatment of Chinese cabbage with ASC reduced the population of artificially inoculated Escherichia coli O157:H7, Salmonella Enteritidis, Staphylococcus aureus, and Listeria monocytogenes by 2.4 log CFU/g. The sanitation efficacy of ASC was 1.6 log CFU/g higher than that of distilled water washing. The viable cell counts of all pathogenic bacteria tested remained constant during 8 days of storage at 10 degrees C for both washing treatments, with the exception of L. monocytogenes, whose viable cell counts increased gradually with time for both treatments. No significant differences in color, odor, taste, and texture in raw leaves were observed after the ASC wash compared with after the distilled water wash. These results indicate that prewashing with ASC could control bacterial growth in lightly fermented Chinese cabbage without changing the product quality. PMID- 15895734 TI - Growth and toxin production by Clostridium botulinum in steamed rice aseptically packed under modified atmosphere. AB - Sales and consumption of ready-to-eat aseptic steamed rice products have increased manyfold in Japan over the past 10 years. To determine the safety of steamed rice (water content 60%, pH 6.5) aseptically packaged under modified atmosphere, challenge studies were performed using a mixture of Clostridium botulinum proteolytic strains (five strains of type A and five strains of type B). Atmospheric conditions of 0 and 15% oxygen (with 5% CO2 and 5% N2 as the balance) were used. No neurotoxins were detected, and organoleptically acceptable conditions persisted for 24 weeks at 15% oxygen conditions. However, botulinum neurotoxin was found in one of three samples at 12 weeks and in one of two samples at 24 weeks at 0% oxygen and 30 degrees C. When samples were inoculated with C. botulinum with amylase (0% oxygen), neurotoxin and sample spoilage was detected after only 1 week of storage. Challenge studies using proteolytic strains of C. botulinum mixed with Bacillus subtilis (amylase formers) also were performed with atmosphere conditions of oxygen at 0, 5, 10, and 15% (with 5% CO2 and 5% N2 as the balance). Under 10 and 15% oxygen conditions, neurotoxin was not detected after 1 week of storage, but sample spoilage was detected after the same period. Under 0% oxygen conditions, neurotoxin was detected at 1 week, but the sample remained organoleptically acceptable even after 2 weeks of storage. Both neurotoxin and sample spoilage were detected at 1 week of storage under 5% oxygen conditions. Based on these results, cocontamination of amylase-producing Bacillus with C. botulinum would increase the risk of foodborne botulism when aseptic rice samples are packed under low-oxygen conditions (<5%). Therefore, to ensure the safety of these products, packing under atmospheric containing more than 10% oxygen is recommended. PMID- 15895735 TI - Antimicrobial activity of nisin adsorbed to surfaces commonly used in the food industry. AB - The adsorption isotherms of nisin to three food contact surfaces, stainless steel, polyethyleneterephthalate (PET), and rubber at 8, 25, 40, and 60 degrees C, were calculated. For all surfaces, the increase in temperature led to a decrease in the affinity between nisin and the surface. The rubber adsorbed a higher amount of nisin (0.697 microg/cm2) in comparison with PET (0.665 microg/cm2) and stainless steel (0.396 microg/cm2). Adsorption of nisin to the stainless steel surface described L-2 type curves for all temperatures assayed. However, for PET and rubber surfaces, the isotherms were L-2 type (at 40 and 60 degrees C) and L-4 type curves (at 8 and 25 degrees C). Nisin retained its antibacterial activity once adsorbed to the food contact surfaces and was able to inhibit the growth of Enterococcus hirae CECT 279 on Rothe agar medium. The attachment of three Listeria monocytogenes strains to the three surfaces was found to be dependent on the surface, the strain, and the initial bacterial suspension in contact with the surface. The adsorption of Nisaplin on surfaces reduced the attachment of all L. monocytogenes strains tested. The effect of PET based bioactive packaging in food was very encouraging. When applied to a food system, nisin-adsorbed PET bottles reduced significantly (P < 0.05) the levels of the total aerobic plate counts in skim milk by approximately 1.4 log units after 24 days of refrigerated storage (4 degrees C), thus extending its shelf life. PMID- 15895736 TI - Influence of packaging conditions on natural microbial population growth of endive. AB - The influence of three packaging conditions, i.e., unmodified atmosphere packaging (UAP), passive modified atmosphere packaging (MAP), and active MAP, on the natural microbial population growth of endive was investigated at 20 degrees C. For UAP, endive was placed in macroperforated oriented polypropylene pouches that maintained gas composition close to that of air (21 kPa O2 and 0 kPa CO2) but also limited superficial product dehydration. For MAP, endive was placed in low-density polyethylene pouches that induced a 3 kPa O2 and 5 kPa CO2 equilibrium atmosphere composition. Steady state was reached after 25 h of storage with an oxygen absorbing packet (active MAP) compared with 100 h without the packet (passive MAP) and was maintained for 200 h. After 312 h of storage, both active and passive MAP reduced total aerobic mesophile, yeast, and mold population growth compared with endive in UAP. Active MAP accelerated and improved the inhibition of Pseudomonas spp. and Enterobacteriaceae, respectively, probably because of the rapid O2 depletion during the transition period. A shift in the Enterobacteriaceae subpopulation from Rhanella aquatilis to Enterobacter agglomerans was observed for both passive and active MAP. PMID- 15895737 TI - Effect of milk inoculation with bacteriocin-producing lactic acid bacteria on a Lactobacillus helveticus adjunct cheese culture. AB - The effect of eight strains of lactic acid bacteria (two strains of Enterococcus, one strain of Lactobacillus, and five strains of Lactococcus, which produce enterocin AS-48, enterocin 607, nisin A, nisin Z, plantaricin 684, lacticin 481, or nisin Z plus lacticin 481) on acid production and proteolytic activity of Lactobacillus helveticus LH 92 (a highly peptidolytic strain used as an adjunct in cheese making) was evaluated in mixed cultures in milk. Acid production by mixed cultures depended on the sensitivity of L. helveticus LH 92 to the different bacteriocins and on the acidification rates of bacteriocin-producing strains. Proteolysis values of mixed cultures were, in all cases, lower than those of L. helveticus LH 92 single culture (control). Cell-free aminopeptidase activity values after 9 h of incubation did not increase in the presence of enterocin producers or the nisin A producer, whereas in the presence of the nisin Z producer, cell-free aminopeptidase activity was, at most, 3.7-fold greater than the control value. In mixed cultures with the plantaricin producer, a progressive lysis of L. helveticus LH 92 took place, with cell-free aminopeptidase activity values after 9 h being, at most, 10.5-fold greater than the control value. The highest cell-free aminopeptidase activity values after 9 h were recorded in the presence of lacticin 481 producers, with the values being, at most, 25.1-fold greater than the control value. L. helveticus LH 92 was extremely sensitive to small variations in the concentration of the inoculum of the nisin Z plus lacticin 481 producer, with there being a narrow optimum for the release of intracellular aminopeptidases. Plantaricin and lacticin 481 producers seemed the most promising strains to be combined with L. helveticus LH 92 as lactic cultures for cheese manufacture,because of the accelerated release of intracellular aminopeptidases. PMID- 15895738 TI - Production of bacteriocin-like inhibitory compounds by human fecal Bifidobacterium strains. AB - Acid- and bile-resistant Bifidobacterium strains were isolated from human feces and identified by genus-specific PCR and randomly amplified polymorphic DNA PCR. Twenty-four different strains were screened for possible production of proteinaceous antimicrobial compounds by assaying the inhibitory effects of their neutralized culture supernatants. Six Bifidobacterium strains (BIR-0304, BIR 0307, BIR-0312, BIR-0324, BIR-0326, and BIR-0349) were selected on the basis of their broad inhibitory spectra. These strains were active against gram-positive and gram-negative bacteria and yeasts relevant to food safety and human health. The antagonistic effects of the six selected Bifidobacterium strains were related to bacteriocin-like compounds, which were active at pH values between 3 and 10, stable at 100 degrees C for 10 min, resistant to alpha-amylase and lipase A, but sensitive to proteinases (trypsin, proteinase K, protease A, pepsin, and cathepsin B). The molecular masses of the antimicrobial compounds produced by Bifidobacterium BIR-0312 and BIR-0324 were in the range of 10 to 30 kDa, and those of the compounds produced by Bifidobacterium BIR-0304, BIR-0307, BIR-0326, and BIR-0349 were less than 10 kDa. All Bifidobacterium strains produced maximum antimicrobial activities in the late logarithmic phase of growth and in the presence of Tween 80. These results confirm that the synthesis of bacteriocin like inhibitory compounds is a key factor in the in vitro inhibition of pathogen and spoilage bacteria by Bifidobacterium strains. PMID- 15895739 TI - Thermal properties and granulometry of dried powders strongly influence the effectiveness of heat treatment for microbial destruction. AB - The thermal treatment of Saccharomyces cerevisiae cells, which were homogeneously incorporated into dried wheat flour particles, was performed for various particle radii (0.8 to 1.6, 1.6 to 2.8, 2.8 to 3.2, and 5 mm) and for an initial water activity of 0.20. A new high-temperature short-time process developed by our laboratory for powder decontamination was used at 150, 200, and 250 degrees C for 5 to 30 s, and significant destruction of up to a 6.7-log reduction, depending on treatment conditions and granule size, was achieved. This study confirms the strong influence of granulometry on the microbial destruction of homogeneously contaminated powdered products. Moreover, a thermal model was developed that takes into account the thermal properties of each component, the variations during heat treatment, and the energy required for phase change. This model provides a tool for predicting yeast destruction. PMID- 15895740 TI - Collaborative evaluation of a fluorometric method for measuring alkaline phosphatase activity in cow's, sheep's, and goat's milk. AB - Pasteurization of raw milk was introduced to extend product shelf life and destroy pathogens. The measurement of alkaline phosphatase (ALP) activity has been used as an indicator of proper pasteurization in dairy products for more than 65 years. This study was undertaken to evaluate six different fluid dairy products at lower phosphatase levels than previously verified using the Fluorophos Test System, a sensitive and precise method for ALP activity detection. Thirteen laboratories participated in this collaborative, international study to evaluate the fluorometric test at 20, 40, 100, 350, and 500 mU/liter and extend the scope of the method to include milk from not only cows but also goats and sheep. Initially, the statutory level of ALP measured fluorometrically was set to equivalent levels of colorimetric test standards (500 mU/liter). The European Union recently announced its intention of lowering the legal limit from 500 to 350 mU/liter and, in addition, setting a target value of 100 mU/liter, which if exceeded would trigger an investigation into the pasteurizer plant performance. At 500 mU/liter of ALP, this trial generated relative standard deviation of repeatability values of 6.48, 5.69, and 1.74% and relative standard deviation of reproducibility values of 14.66, 13.30, and 5.33% for all cow's, sheep's, and goat's milk samples, respectively. Data from this study are comparable to data from previous studies and indicate the suitability of the Fluorophos Test System method for measuring ALP activity in milk from cows, sheep, and goats not only at the current European statutory level of 500 mU/liter but also at much lower levels. PMID- 15895741 TI - Effect of water activity and temperature on growth and the relationship between fumonisin production and the radial growth of Fusarium verticillioides and Fusarium proliferatum on corn. AB - The two major fumonisin-producing Fusarium species are Fusarium verticillioides and Fusarium proliferatum. The growth and fumonisin production of these two isolates on corn was studied at water activities (a(w)) between 0.860 and 0.975 and at temperatures between 15 and 30 degrees C. Growth rates (g, mm/day) were obtained by linear regression during the linear phase of growth. In general, growth rates for both isolates increased significantly (P < 0.05) with increases in a(w) and temperature. Both fumonisin production and radial growth (mycelial development) for both isolates increased with a(w) at all temperatures investigated, but the effect of temperature on this relationship was not obvious. The effect of temperature on fumonisin production at high a(w) values optimal for growth was only marginal, whereas at lower a(w) values the effect of temperature was more pronounced, with more fumonisin production occurring at temperatures not optimal for growth. The optimum temperature for fumonisin production was between 15 and 25 degrees C. For F. proliferatum, the optimum temperature for growth at all a(w) values, 30 degrees C, resulted in the poorest fumonisin production. For both isolates, the slowest initial rate of fumonisin production was at 15 degrees C, the temperature at which the slowest growth rates were obtained. PMID- 15895742 TI - Microbiological and aflatoxin evaluation of Brazil nut pods and the effects of unit processing operations. AB - Harvesting of Brazil nuts not only helps to preserve the Amazon rainforest but also provides income to individuals who would otherwise have little means of making a livelihood. Recently, the European Community has tightened the quality requirements for Brazil nuts, particularly with regard to aflatoxin levels and microbiological contamination. The objectives of this research were to gain a better understanding of the origin of aflatoxins on Brazil nuts and to microbiologically evaluate some of the operations involved in processing. In this regard, five Brazil nut pods were aseptically picked from trees located in each of three concessions of the Peruvian Amazon rainforest (Madre de Dios province). The exteriors of the pods and the nuts were examined for yeast and molds, including Aspergillus flavus and Aspergillus parasiticus, and for bacteria, including Salmonella and Escherichia coli. Brazil nuts obtained from various commercial process operations located in Peru were similarly evaluated. Exteriors of all Brazil nut pods did not contain A. parasiticus, and only pods from one concession yielded A. flavus isolates. All isolates tested were aflatoxigenic (630 to 915 ppb total aflatoxin). Coliforms, E. coli, and salmonellae were not recovered from any of the pods. Whole, in-shell nuts obtained after opening the pods yielded no A. flavus or A. parasiticus. Aflatoxins were not detected (detection limit 1.75 ppb) in any of the nuts. Whole, in-shell and shelled nuts from various process operations were all positive for A. flavus but negative for E. coli and salmonellae. Soaking of whole, in-shell nuts before cracking or shelling increased coliform numbers, whereas levels of A. flavus decreased. In order to gain a better understanding of the sanitary performance of the unit process operations, additional evaluations should be conducted on product lots processed on different days. Also, the microbiology of product processed from common lots should be followed through the various unit operations and compared. PMID- 15895743 TI - A new procedure for marinating fresh anchovies and ensuring the rapid destruction of Anisakis larvae. AB - The consumption of marinated anchovies is the main route of transmission of anisakiasis in Spain. Because this country is one of the world's major tourist destinations, this traditional food also poses a potential health risk to millions of foreign visitors. Anisakis larvae are not destroyed by the traditional marinating procedure, and alternative methods, such as long-term storage in brine, freezing, or hydrostatic pressure treatment, all present major difficulties. In this study, we used high food-grade acetic acid concentrations (10, 20, 30, and 40% [vol/vol] in line with the quantum satis rule) to destroy these larvae rapidly, and we report data on the survival of Anisakis larvae exposed directly to different marinades and when the larvae are placed under the fish musculature. The percentage of salt and acetic acid in the fish tissue water phase was also determined. A marinating procedure is proposed that ensures the rapid death of Anisakis through the use of strong acetic acid concentrations. Posttreatment washes with water reduce these to levels acceptable to consumers. The sensory characteristics of the product were shown to be satisfactory. The actual selection of an acetic acid concentration for marinating depends on costs and the processing time available. The physiological stress of the larvae exposed to the different marinades was determined by measuring the levels of their stress proteins. The latter are good indicators of injury and might reflect the infectivity of larvae. In addition, we also used a rat model to determine the infectivity of larvae considered microscopically dead. PMID- 15895744 TI - A comparison of sample weight and culture methods for the detection of Salmonella in pig feces. AB - Five protocols were compared to determine the combined effects of different sample weights and culture methods for the recovery of Salmonella from 310 pig cecal samples taken in abattoirs as part of the Canadian Integrated Program for Anti-microbial Resistance Surveillance. Sample weights evaluated were 1 and 10 g. Culture methods used with each sample weight were modified semisolid Rappaport Vassiliadis agar (MSRV) and brilliant green agar with sulfa and novobiocin (BGSN) and xylose-lysine-tergitol-4 agar (XLT4). A preliminary sample preparation step in saline was also evaluated using a 10-g sample and MSRV. The Salmonella recovery rate varied from 20% for the saline MSRV 10-g protocol to 32% for the MSRV 10-g and the BGSN-XLT4 10-g protocols. A good agreement (K > 0.8) was observed between pairs of protocols except whenever the saline MSRV 10-g and the MSRV 1-g protocols were compared. Larger samples (10 g) yielded higher detection of Salmonella than 1-g samples for the MSRV protocol (32 versus 25%), whereas the differences were not statistically significant for the BGSN-XLT4 protocols. Protocols using the BGSN-XLT4 agar yielded higher detection rates of Salmonella compared with MSRV with 1-g samples (30 versus 25%), whereas it was equivalent with 10-g samples. Considering a greater recovery rate, the ease of use, and a better time and resource efficiency, the MSRV 10-g protocol was therefore adopted by the Canadian Integrated Program for Antimicrobial Resistance Surveillance. PMID- 15895745 TI - Contamination of Salmonella in retail meats and shrimps in the Mekong Delta, Vietnam. AB - From March 2000 to September 2001, 608 samples of retail meat (136 pork, 70 beef, 202 chicken, and 200 ducks) and 110 samples of retail shrimp from six provinces of the Mekong Delta in Vietnam were collected individually and examined for the prevalence of Salmonella. Of the 718 samples examined, 243 (33.8%) were Salmonella positive. Salmonella was isolated from 69.9% of the pork samples, 48.6% of the beef samples, 21.0% of the chicken meat samples, 22.3% of the duck meat samples, and 24.5% of the shrimp samples. From 261 Salmonella isolates, 24 different serovars were identified. The predominant serovars of the isolates were Salmonella Derby, Salmonella Weltevreden, and Salmonella London in pork; Salmonella Weltevreden, Salmonella London, and Salmonella Dessau in beef; Salmonella Emek, Salmonella Typhimurium, and Salmonella Dessau in chicken meat; Salmonella Lexington, Salmonella Derby, and Salmonella Dessau in duck meat; and Salmonella Weltevreden, Salmonella Tennessee, and Salmonella Dessau in shrimps. Salmonella Bovismorbificans, Salmonella Derby, Salmonella Dessau, and Salmonella Weltevreden were the most common serovars in all the samples examined. These results indicate a high rate of contamination by Salmonella in retail meats and shrimps in the Mekong Delta, Vietnam. PMID- 15895746 TI - Differences in heat resistance among pathogenic Yersinia enterocolitica depended on growth temperature and serotype. AB - To gain a better understanding about the effect of growth temperature on heat resistance of Yersinia enterocolitica, we determined decimal reduction times at 60 degrees C (D60-values) for O:3; O:5,27; O:8; and O:9 strains harboring virulence plasmid coding for Yersinia outer membrane protein and experimentally virulence plasmid-deleted strains after they were grown to stationary phase at 7, 25, or 37 degrees C. Bacteria were inoculated into Trypticase soy broth and were incubated at several temperatures. D60-values of O:3; O:5,27; and O:8 strains were larger when they were grown at 37 degrees C than at 7 or 25 degrees C, despite the presence or absence of virulence plasmids. However, similar D60 values were observed in O:9 strains, despite growth at 7, 25, or 37 degrees C. The results indicate two types of Y. enterocolitica strains, growth temperature dependent and -independent, and a Yersinia outer membrane protein that is not directly involved in growth temperature-dependent heat resistance. PMID- 15895747 TI - Development of a quantitative real-time PCR method for estimation of the total number of Vibrio parahaemolyticus in contaminated shellfish and seawater. AB - A real-time PCR method targeting the toxR gene of Vibrio parahaemolyticus was developed to quantify the number of V. parahaemolyticus cells, including those of both the hemolysin-producing and nonproducing strains. The specificity of the primer and probe set was confirmed using 25 strains of V. parahaemolyticus and 30 strains of other microbial species. We determined the threshold cycle number using the real-time PCR and the number of V. parahaemolyticus cells by plate count using serially diluted pure culture and developed a standard curve for quantification. Standard curves for V. parahaemolyticus in seawater and seafood were established using artificially inoculated samples. The threshold cycle number and the number of V. parahaemolyticus cells were correlated with 10(1) to 10(7) CFU/ml in pure culture, seawater, and shellfish homogenate. The real-time PCR method developed in this study was compared with the most-probable-number method in seafood samples that were naturally contaminated. The differences in the number of V. parahaemolyticus cells as determined by the culture method and the PCR method were less than 10-fold. PMID- 15895748 TI - Head space sensor array for the detection of aflatoxin M1 in raw ewe's milk. AB - A novel screening method was developed for simple and rapid detection of aflatoxin M1 contamination in raw ewe's milk samples without the need for sample pretreatment. The method was based on the use of a commercial head space sensor array system constituted by 12 metal oxide semiconductor sensors, 10 metal oxide semiconductor field-effect transistor sensors, and a pattern recognition software. Twenty-four raw milk samples collected from two different groups of ewes fed with a formulated feed that contained increasing amounts of aflatoxin B1 and six noncontaminated ewe's milk samples were analyzed. The results obtained by using the head space sensor array, processed by statistical methods, made it possible to group the samples according to the presence or the absence of aflatoxin M1. Sample classification was in complete agreement with the aflatoxin M1 content measured by an enzyme-linked immunosorbent assay procedure. This is the first report, to our knowledge, of detection of aflatoxin M1 in ewe's milk by a multisensor array. PMID- 15895749 TI - Detection of Cryptosporidium parvum oocysts on fresh vegetables and 1 herbs using antibodies specific for a Cryptosporidium parvum viral antigen. AB - The purpose of this study was to determine if the viral symbiont of Cryptosporidium parvum (CPV) sporozoites could be used as a target for sensitive detection of the parasite in food samples. Polyclonal sera specific to the recombinant viral capsid protein (rCPV40) was used in a dot blot hybridization assay to detect oocysts recovered from green onions and cilantro. Small batches of chopped green onions and cilantro leaves were artificially contaminated with three different concentrations of oocysts: 10(6), 10(2), and 10(1). rCPV40 was superior in detecting oocysts compared with other antibodies directed toward total oocyst protein and oocyst surface antigens. This study provides evidence that CPV is an excellent target for sensitive detection of C. parvum oocysts in foods. PMID- 15895750 TI - Fate of eprinomectin in goat milk and cheeses with different ripening times following pour-on administration. AB - The distribution of eprinomectin in goat milk and cheeses (cacioricotta, caciotta, caprilisco) with different ripening times following a pour-on administration at a single dose rate (500 microg/kg of body weight) and a double dose rate (1,000 microg/kg of body weight) to goats with naturally occurring infections of gastrointestinal nematodes was studied. Milk residues of eprinomectin reached a maximum of 0.55+/-0.18 microg/kg and 1.70+/-0.31 microg/kg at the single and double doses, respectively. The drug concentrations decreased progressively until the fifth day after treatment, when they were less than the detection limit at both dose rates. The eprinomectin levels measured in all cheese types (both treatments) were higher than those recovered in milk at all the sampling times. In caciotta cheeses, the eprinomectin residues levels were constantly higher than other cheeses. With the exception of cheeses made with milk the first day after treatment, eprinomectin concentrations were nearly constant up to the fourth day then decreased by the fifth and sixth days after treatment. In all cases, at both the single and double dosages, the maximum level of eprinomectin residues in goat milk and cheeses remained below the maximum residual level of 20 microg/liter permitted for lactating cattle. PMID- 15895751 TI - Bacteriophage control of foodborne bacteriat. AB - Bacteriophages are measurable components of the natural microflora in the food production continuum from the farm to the retail outlet. Phages are remarkably stable in these environments and are readily recovered from soil, sewage, water, farm and processing plant effluents, feces, and retail foods. Purified high-titer phage lysates have been used for the species-specific control of bacteria during the pre- and postharvest phases of food production and storage. For example, the inhibition of the phytopathogens Erwinia amylovara and Xanthomonas campestris has reduced the incidence of diseases such as fire blight in apples and bacterial spot of tomato and peaches. Research on preslaughter treatment of food animals has demonstrated phage control of salmonellosis in chickens, enteropathogenic Escherichia coli infections in calves, piglets, and lambs, and E. coli O157:H7 shedding by beef cattle. Phages have also been applied to control the growth of pathogens such as Listeria monocytogenes, Salmonella, and Campylobacter jejuni in a variety of refrigerated foods such as fruit, dairy products, poultry, and red meats. Phage control of spoilage bacteria (e.g., Pseudomonas spp. and Brochothrix thermosphacta) in raw chilled meats can result in a significant extension of storage life. Phage biocontrol strategies for food preservation have the advantages of being self-perpetuating, highly discriminatory, natural, and cost effective. Some of the drawbacks of biopreservation with phages are a limited host range, the requirement for threshold numbers of the bacterial targets, phage resistant mutants, and the potential for the transduction of undesirable characteristics from one bacterial strain to another. Most research to date has involved experimentally infected plants and animals or artificially inoculated foods. This technology must be transferred to the field and to commercial environments to assess the possibility of controlling natural contaminants under more realistic production and processing conditions. PMID- 15895752 TI - Proposed conditions for coverage directed toward dialysis facilities. PMID- 15895753 TI - "The birth of a clinic"? The IMS dispensary in Gyantse (Tibet), 1904--1910. PMID- 15895754 TI - "A mere matter of rock": organized labour, scientific evidence and British government schemes for compensation of silicosis and pneumoconiosis among coalminers, 1926--1940. PMID- 15895755 TI - Breastfeeding and health professionals in Britain, New Zealand and the United States, 1900--1970. PMID- 15895756 TI - Spectacle and secrecy: press coverage of conjoined twins in 1950s Britain. PMID- 15895757 TI - The anatomist by Thomas Rowlandson (1756--1827): the play's the thing. PMID- 15895758 TI - Presidential address: The state of the union of vascular surgery, 2004. PMID- 15895759 TI - Hemorrhagic complications during long-term postoperative warfarin administration in patients undergoing lower extremity arterial bypass surgery. AB - Lower extremity bypass procedures restore function and prevent amputation in many patients with severe peripheral arterial occlusive disease. The regular postoperative use of aspirin offers the dual benefit of extending bypass patency and patient survival. Previous trials of adjunctive oral anticoagulant therapy with warfarin have infrequently combined warfarin with aspirin. We hypothesized that the addition of oral anticoagulant therapy would further enhance the benefits of aspirin but may increase the risk of clinically important bleeding. Eligible patients (N = 831) scheduled for elective lower extremity arterial bypass surgery were randomized to receive either warfarin plus aspirin (WA) (n = 418) or aspirin alone (n = 413). At monthly intervals, the warfarin dose was adjusted to a target international normalized ratio (INR) of 1.4 to 2.8; both groups received aspirin (325 mg/d). The end point of major hemorrhagic events, defined as intracranial hemorrhage or bleeding that required intervention, is reported, and INR values and compliance with warfarin therapy are presented. Major hemorrhagic events occurred more frequently in the WA group (35 in the WA group vs 15 in the aspirin group; p = .02) during a mean follow-up of 38 months. In the WA group, an intracranial hemorrhage occurred in six patients (two had an INR > 3.0), of whom four died; one subdural hemorrhage occurred in the aspirin group. Transfusions and interventions for bleeding were more frequent in the WA group, as were minor bleeding events. Of the 8,946 INR determinations, 58% were in the target range, whereas a higher value occurred in 10% and a lower value in 32%. Compliance with warfarin was maintained in 65% of the patients after the first year of observation. In patients with elective lower extremity bypass procedures, the postoperative adjunctive use of warfarin with aspirin increased the risk of major hemorrhagic events. Most of these events occurred when the INR was in the target range. PMID- 15895760 TI - Carotid stent mobility with regard to head movements: in vitro analysis. AB - Given that considerable motion of the carotid artery is present during head movements, we hypothesized that a flexible stent with low torsion might be favorable to avoid stress imparted to the stent and carotid artery. Therefore, we evaluated the flexibility of different expanded carotid stents before and after deployment in a carotid artery in vitro. Subsequently, we evaluated torsion of the bare expanded stents. Five stents (Wallstent [Boston Scientific Corp., Natick, MA], Acculink [Guidant Corp., Indianapolis, IN], Precise [Cordis Corp., Johnson & Johnson Company, Warren, NJ], Carotid SE [Medtronic AVE, Santa Rosa, CA], and Protege [EV3, Plymouth, MN]) were tested. Flexibility was determined using a three-point bend test recording the bending load (BL) in grams required to flex the stent 25 degrees. Increased BL implies decreased flexibility. Torsion was measured by recording the rotation load (RL) in grams required to rotate the stents 30 degrees along its axis. Increased RL implies increased torsion. In the bare expanded state, the median BL was 6 g (range 1-22 g). The BL increased to 38 g (range 20-41 g) after deployment in a carotid artery, with the Carotid SE (21 g) and Wallstent (36 g) showing significantly lower BL (p < .0001 and p = .0016, respectively). Overall, the RL was 11 g (range 1-76 g). Significantly higher RL was required to rotate the Wallstent (73 g) and Precise (20 g) stents (p < .0001). The flexibility of the currently used stents decreases after deployment in a carotid artery irrespective of its flexibility in the bare state. Two stents showed increased torsion compared with the other stents. Limitations in both flexibility and torsion might influence the long-term performance of carotid angioplasty stenting. PMID- 15895761 TI - Venous thromboembolism: regional differences in the nationwide inpatient sample, 1993 to 2000. AB - Venous thromboembolism (VTE) is a costly complication of hospitalization. The sequelae make it a concern for public health planners. The Nationwide Inpatient Sample (NIS) contains data for hospital discharges in the United States. These data were reviewed to determine their suitability for health policy planning. International Classification of Diseases, Ninth Revision, Clinical Modification codes for VTE were applied to the NIS data. The sample was queried for demographic information, mortality, length of hospital stay, diagnosis, and treatment. The rates were standardized for geographic region and disease acuity. Statistical analysis included descriptive reporting of means and event rates; analysis of variance and logistic regression were used for regional effects and modeling of mortality. Between 1993 and 2000, 636,814 discharges involved VTE (1.2%). This rate was consistent over time and within regions. Regional differences existed in the acceptance of new technology and hospital charges. Mortality varied from 6.3% (Midwest) to 7.9% (Northeast) and was associated with admission type, comorbidities, pulmonary embolism, and discharge from the Northeast region. White race, chronic venous insufficiency, and female gender were protective variables. The NIS data report a consistent mortality rate despite improved therapy. Regional diagnostic, treatment, and economic differences exist. The data are useful for the purposes of public health care planning and stimulating clinical trial questions. PMID- 15895762 TI - Outcome of concomitant renal artery reconstructions in patients with aortic aneurysm and occlusive disease. AB - The question remains as to whether patients presenting with aortoiliac occlusive disease (AIOD) or abdominal aortic aneurysms (AAAs) have similar outcomes when concomitant renal artery reconstructions are performed. In this study, we analyzed our experience with simultaneous aortic and renal reconstructions using a retroperitoneal approach. Over a 5-year period, all patients with either AAAs > 5 cm or symptomatic AIOD who were found to have high-grade renal artery stenosis and who underwent aortic reconstructions with concomitant renal revascularization were analyzed through our vascular surgery registry. Morbidity and mortality were quantitatively evaluated. Data were analyzed using the chi-square test. A total of 1,133 patients with AAA (n = 832) and AIOD (n = 301) underwent aortic reconstructions. Two hundred thirty-one patients had 283 concomitant renal revascularizations, including bypass, reimplantation, and endarterectomy, for high-grade (> 70%) renal artery stenosis via a left retroperitoneal approach. The mortality rate of AAA repair with and without renal revascularization was 2.3% (4/178) and 1.5% (10/654), respectively, and that of aortobifemoral bypass for AIOD with and without renal revascularization was 5.7% (3/53) and 2.8% (7/248), respectively. Of the 7 deaths in patients requiring aortic and renal reconstructions, 4 occurred in patients with bilateral renal revascularization. Transient renal insufficiency, ischemic colitis, and cardiopulmonary failure occurred in 5.6%, 2.2%, and 9.6% of patients with AAA repair and in 5.7%, 0%, and 9.4% of patients with AIOD. Two patients developed acute occlusion of their renal bypasses; one was successfully revised, whereas the other led to a nephrectomy. In patients with AAAs, AIOD, and high-grade renal artery stenosis, simultaneous aortic and renal reconstructions can be performed through a retroperitoneal approach with a limited and acceptable mortality. With concomitant renal and aortic procedures, patients with AIOD have a higher mortality when compared with those with AAAs, although this difference is not statistically significant. PMID- 15895763 TI - Bilateral external carotid artery aneurysms. AB - A 60-year-old female was found on a physical examination to have bilateral palpable pulsatile neck masses. She denied local pain, cranial nerve compressive symptoms, or symptoms of cerebral ischemia. Duplex ultrasonography demonstrated bilateral 1.5 x 2.2 cm external carotid artery aneurysms. Isolated bilateral external carotid artery aneurysms were confirmed by computed tomography and angiography. The patient has been treated conservatively, and at 4-year follow up, she remains asymptomatic, and the carotid artery aneurysms are unchanged in size. PMID- 15895764 TI - Giant celiac artery aneurysm with associated visceral occlusive disease. AB - Celiac artery aneurysms are rarely seen in clinical practice. We report an unusual case of a large celiac artery aneurysm in a patient with associated visceral occlusive disease who presented with vague abdominal pain and underwent uneventful open surgical repair. PMID- 15895765 TI - In defense of listening. AB - This paper makes a case for listening and its role in establishing and maintaining meaningful psychotherapeutic relationships. Further, it examines those factors that have contributed to the devaluation and current decline of listening in mental health care. These factors include the abuses of managed care, the marketing and misuse of psychotropic medications, the growth of brief, manualized, empirically supported treatments (ESTs), and the lack of appreciation for unconscious processes and countertransference in the appeal, development and delivery of these interventions. PMID- 15895766 TI - The meanings of medicating: pills and play. AB - The author uses Winnicott's theory involving transitional objects, the therapeutic value of play, and the vital role of danger to elucidate the psychotherapeutic aspects of medication visits. A case vignette illustrates growth of the possibility of play in the course of visits whose overt agenda is "meds only." PMID- 15895767 TI - The use of the 'corrective emotional experience' and the search for the bad object in psychotherapy. AB - In this article the classical concept and the use of the 'corrective emotional experience' in psychotherapy is described. This concept is not widely accepted within the standpoint of contemporary psychoanalytic psychotherapy and object relations theories. It has an unfavorable reputation. However, this paper does not advocate discarding this therapeutic idea and action because it does have therapeutic value. The case is made that the search for the bad object, and patient's reenactment of early experiences as they had been, is a corrective emotional experience in that it is a reparative experience. Furthermore, the search for the bad object is necessary for attachment. In addition, the point is made that patients find and create the object, therefore, the corrective emotional experience must be viewed as something that patients find rather than what therapists do. PMID- 15895768 TI - The space between love and not touching in psychotherapy. AB - The present paper discusses the advent and significance of erotic feelings in psychotherapy based on the intersubjective theory. It briefly reviews the coalescence of the tradition of avoidance of physical contact in psychotherapy, and the classical and contemporary approaches to erotic transference. A clinical case is presented in an attempt to expand the significance attributed to erotic feelings in therapy and ways of relating to it: in their intrapersonal and interpersonal meanings and in the dialectics between them. I will later discuss the father figure in feminine development and its transferential and metaphoric meaning against the background of a clinical case, and the importance of the erotic component in the processes of change and development. The paper includes mythological metaphors illustrating the issues in a cultural perspective that constitutes the traditional framework of thinking relevant to the issues which are the subject of this paper. PMID- 15895770 TI - Agreeing to disagree. PMID- 15895769 TI - An outcome of psychodynamic psychotherapy: a case study of the change in serotonin transporter binding and the activation of the dream screen. AB - We explored the outcome of psychodynamic psychotherapy of a female patient with major depression using clinical evaluation and serotonin transporter (SERT) binding assessed with [123I]nor-beta-CIT SPECT. The psychotherapy process was analyzed with special emphasis on the change that was recognized in the dreaming process. The activation of the dream screen in transference seemed to form a turning point during the psychotherapy. Normalization of SERT binding at the midbrain level was found on 12-month follow-up. Major alleviation of depressive symptoms assessed by rating scales was evident only six months after SERT normalization. PMID- 15895771 TI - Smoking-cessation support. PMID- 15895772 TI - Smoking-cessation support. PMID- 15895773 TI - Clinical utility of autoantibodies directed against TSH-R. PMID- 15895774 TI - Assess your sharps injury prevention program. PMID- 15895775 TI - Inland Hospital debuts and no.1 with its "star" performers. PMID- 15895776 TI - SCC's bottom line reflects the value of training. PMID- 15895777 TI - The plus and minus sides of Rh transfusions. PMID- 15895778 TI - Chronic cough as a sign of laryngeal sensory neuropathy: diagnosis and treatment. AB - Chronic cough is often attributed to reflux, postnasal drip, or asthma. We present 28 patients who had chronic cough or throat-clearing as a manifestation of sensory neuropathy involving the superior or recurrent laryngeal nerve. They had been identified as having sudden-onset cough, laryngospasm, or throat clearing after viral illness, surgery, or an unknown trigger. Cough and laryngospasm were the most common complaints. Seventy-one percent of the patients had concomitant superior laryngeal nerve or recurrent laryngeal nerve motor neuropathy documented by laryngeal electromyography or videostroboscopy. After a negative workup for reflux, asthma, or postnasal drip, these patients were treated with gabapentin at 100 to 900 mg/d. Symptomatic relief was achieved in 68% of the patients. Sensory neuropathy of the recurrent laryngeal nerve or superior laryngeal nerve should be considered in the workup for chronic cough or larynx irritability. Symptomatic management of patients with cough and laryngospasm due to a suspected sensory neuropathy may include the use of antiseizure medications such as gabapentin. PMID- 15895779 TI - Altered laryngeal sensation: a potential cause of apnea of infancy. AB - Delayed maturation of respiratory control of breathing and the laryngeal adductor reflex (LAR) are commonly implicated in infant apnea. A swallow response occurs to remove the stimulus from the pharynx to prevent aspiration once the glottis reopens. Induction of apnea by poorly cleared endogenous upper airway secretions has been postulated to be a potential cause of infant apnea. Our purpose was to determine whether alteration in the LAR, an indicator of laryngeal sensation, and the presence of secretions influenced the responsiveness of the LAR in infants with apnea. The LAR was induced in 20 infants with apnea (median gestational age, 36.5 weeks) by application of air pulses of controlled duration (50 ms) and intensity (2.5 to 10 mm Hg) to the aryepiglottic fold. Twenty infants evaluated for upper respiratory tract anomalies were used as a comparison group (median gestational age, 39 weeks). The infants with apnea required higher-intensity stimuli (p < .001) to induce the LAR (6.2 +/- 1.6 mm Hg) than did the comparison group (4.3 +/- 1.0 mm Hg) and demonstrated poorer clearance of secretions (p < .001). These findings were significant even when we adjusted for postconceptional age at the time of the test (p = .007). The findings of this study suggest that decreased laryngeal sensitivity results in poor endogenous secretion clearance and that it may induce a prolonged glottic closure event to prevent aspiration. This closure may play a role in infant apnea. PMID- 15895780 TI - Suppression of thyroarytenoid muscle responses during repeated air pressure stimulation of the laryngeal mucosa in awake humans. AB - Repeated stimulation of the laryngeal mucosa occurs during speech. Single stimuli, however, can elicit the laryngeal adductor response (LAR). Our hypothesis was that the LAR to repeated rapid air pressure stimuli is centrally suppressed in humans. Hooked-wire electrodes were inserted into the thyroarytenoid and cricothyroid muscles on both sides and into the posterior cricoarytenoid muscle on one side. Pairs of air puff stimuli were presented to the mucosa over the arytenoids at pressure levels three times threshold with interstimulus intervals from 250 to 5,000 ms. Bilateral thyroarytenoid responses occurred at around 150 ms to more than 70% of the initial stimuli. With repeated presentation at intervals of 2 seconds or less, the percent occurrence decreased to less than 40% and response amplitudes were reduced by 50%. Central suppression of adductor responses to repeated air puff stimuli may allow speakers to produce voice without eliciting reflexive spasms that could disrupt speech. PMID- 15895781 TI - Protective glottic closure: biomechanical effects of selective laryngeal denervation. AB - Glottic closure constitutes the primary mechanism for prevention of intradeglutitive and postdeglutitive aspiration. Laryngeal paralysis therefore exerts a considerable impact on deglutition, yet little is understood regarding the biomechanical effects of selective denervation on the laryngeal protective function. We measured the glottic closing force (GCF) in each of 6 male, 40-kg Yorkshire pigs 1) after selective unilateral superior laryngeal nerve (SLN) section; 2) after selective unilateral recurrent laryngeal nerve (RLN) section; and/or 3) after combined SLN-RLN section as both right and left SLNs were simultaneously stimulated to evoke the glottic closure response. Stimulation was provided through an oscilloscope with bipolar platinum-iridium electrodes, and the GCF was measured with a pressure transducer positioned between the vocal cords. Six repetitive measures of GCF were obtained before nerve section, and 6 after nerve section, in each subject. Unilateral SLN section reduced the GCF to 54.14% of control, RLN section reduced the GCF to 23.39% of control, and combined SLN-RLN section reduced the GCF to 22.67% of control. These findings underscore the profound differential effects exerted by isolated lesions on the glottic closure function. PMID- 15895782 TI - Evaluation of middle and distal esophageal diverticuli with transnasal esophagoscopy. AB - To emphasize the utility of office-based transnasal esophagoscopy (TNE) in the evaluation of patients with swallowing complaints, we present 2 case reports and a review of the literature. The 2 patients both presented with complaints of chronic dysphagia, globus sensation, and a sensation of "food sticking" with swallowing. The patients were counseled to undergo esophagoscopy. Informed consent was obtained. The nasal cavities and pharynx were anesthetized with topical 4% lidocaine hydrochloride solution. Transnasal esophagoscopy was performed. The procedure was well tolerated by the patients. Esophagoscopy revealed diverticuli in various segments of the esophagus, including the midesophageal and distal areas. The cause of the patients' complaints could be well attributed to the endoscopic findings. No morbidity was associated with the TNE examination. A review of the literature concerning office-based TNE was performed. We found no published reports of middle or distal esophageal diverticuli detected on routine office TNE. We conclude that transnasal esophagoscopy is a relatively safe and efficient tool that can be used in the office setting for evaluation of swallowing complaints. PMID- 15895783 TI - Reepithelialization of orthotopic tracheal allografts prevents rejection after withdrawal of immunosuppression. AB - Prior work has demonstrated that immunosuppressed orthotopic tracheal allografts undergo progressive reepithelialization over a 48-day period with recipient derived tracheal epithelium. We hypothesized that reepithelialization of tracheal allografts would prevent rejection after withdrawal of immunosuppression. BALB/c murine tracheal grafts were transplanted orthotopically into either syngeneic or allogeneic C57/BL6 recipients. The recipients were either not immunosuppressed, immunosuppressed with cyclosporine A (10 mg/kg per day) continuously, or immunosuppressed for 48 days and then withdrawn from immunosuppression. The grafts were assessed for acute and chronic rejection 10 days and 50 days after immunosuppression withdrawal. The immunosuppressed allograft recipients maintained a ciliated epithelium acutely and chronically after immunosuppression withdrawal. Ten days after immunosuppression withdrawal, there was a mild cellular infiltrate, which resolved 50 days after withdrawal. Electron microscopy, lymphocyte subpopulation assays, and lamina propria analysis demonstrated that immunosuppression withdrawal did not result in tracheal allograft rejection. In vitro and in vivo assessments did not demonstrate evidence of systemic or local immune tolerance. We conclude that reepithelialization of orthotopic tracheal allografts with recipient-derived mucosa prevents rejection of allograft segments. Tracheal transplantation may require only transient immunosuppression, which can be withdrawn after tracheal reepithelialization. PMID- 15895784 TI - Role of vascular endothelial growth factor-A in recurrent respiratory papillomatosis. AB - Vascular endothelial growth factor-A (VEGF-A) is known to play an important role in the angiogenic response essential for tumor growth in a variety of human and experimental tumors. This study was designed to investigate whether VEGF-A may play a role in the pathogenesis of recurrent respiratory papillomatosis (RRP). A retrospective study with institutional review board approval was performed at a tertiary care medical center on 12 patients with a history of laryngeal RRP. Their ages at the time of initial diagnosis ranged from 19 to 96 months (mean, 56 months). All patients had involvement of right and left true vocal cords. All patients required multiple endoscopic procedures (range, 4 to 66; mean, 12). Normal pediatric larynx samples from 5 autopsy patients were used as controls. Formalin-fixed, paraffin-embedded sections of laryngeal squamous papillomas from the 12 patients with a diagnosis of RRP and the 5 control patients were examined by in situ hybridization for the presence of messenger RNA (mRNA) for VEGF-A and vascular endothelial growth factor receptor 1 (VEGFR-1) and vascular endothelial growth factor receptor 2 (VEGFR-2). The biopsy specimens were from the true vocal cord (N = 10) or subglottis (N = 2) in the patients with RRP and consisted of large sections of larynx including the true vocal cord in the control patients (N = 5). Strong expression of VEGF-A mRNA was noted in the squamous epithelium of papillomas of all 12 patients. Strong expression of VEGFR-1 and VEGFR-2 was noted in the endothelial cells of the underlying vessels in all 12 patients. Neither strong labeling of VEGF-A mRNA nor labeling of its receptors wasnoted in the control patients. We conclude that the angiogenic growth factor VEGF-A is strongly expressed in the epithelium of squamous papillomas in RRP. Also, VEGFR-1 and VEGFR-2 mRNAs are strongly expressed by underlying vascular endothelial cells, suggesting an important role in the pathogenesis of RRP. PMID- 15895785 TI - Acute histologic effects of extraesophageal reflux on vocal fold healing. AB - This study evaluates how extraesophageal reflux affects membranous vocal fold healing in a canine model. We created membranous vocal fold injuries in the animals and randomly assigned them to topical application of acid and pepsin at pH 2 or pH 6 or of normal saline solution every other day for 12 days. The experimental vocal folds were compared to uninjured, control vocal folds from animals painlessly sacrificed for other reasons. Hematoxylin and eosin, fibronectin, and procollagen I staining were performed for histologic analysis. The injured specimens had three times greater cellular infiltrate (p < or = .001, analysis of variance) and twice as much fibronectin and procollagen I (p < or = .001, analysis of variance) as did the specimens from the control animals. No significant differences or trends were identified for cellular infiltrate, fibronectin, or procollagen I within the injured groups (p > .05, Bonferroni t test). Acute wound healing did not appear to be influenced by the presence of acid and pepsin at pH 2 or 6 as compared to saline solution. PMID- 15895786 TI - Regeneration of aged vocal folds with basic fibroblast growth factor in a rat model: a preliminary report. AB - Aged vocal folds have been reported to have dense collagen deposition and decreased hyaluronic acid (HA) in the lamina propria. These characteristics are thought to contribute to vocal problems that occur with age (presbyphonia). To restore better viscoelasticity to aged vocal folds, an intervention that might increase HA and decrease collagen production from aged vocal fold fibroblasts would appear to be a potentially useful approach. Our previous in vitro study has revealed that basic fibroblast growth factor (bFGF) consistently stimulates HA production and decreases collagen production from aged rat vocal fold fibroblasts. The present in vivo study examined the effects of intracordal injection of bFGF into aged rats' vocal folds in terms of restoration of HA and collagen distribution in the lamina propria. We injected bFGF transorally into the lamina propria of (unilateral) vocal folds. The injection was repeated 4 times weekly, and rats were painlessly sacrificed 1 week, 1 month, and 2 months after the final injection. Histologic examination revealed that bFGF significantly increased the HA content of the lamina propria up to 2 months, but showed no effect on collagen, even after 2 months. Because it might take longer for excessive collagen to be degraded, further studies are necessary to clarify the long-term effect on collagen. A drug delivery system for bFGF also needs to be developed to maximize its effect in the future. The present study suggested at least a positive effect of bFGF in restoring the HA content in the aged vocal fold lamina propria. PMID- 15895787 TI - Techniques and outcomes of laryngeal cleft repair: an update to the Great Ormond Street Hospital series. AB - We present an update to the Great Ormond Street Hospital series of laryngeal clefts, describing a further 35 clefts of Benjamin-Inglis types 1 through 3 treated between 1992 and 2003. Associated congenital anomalies were common. Most type 1 and smaller type 2 clefts were repaired endoscopically, whereas larger clefts were repaired through an anterior approach. Increasing use was made recently of a 3-layer repair with an interposition graft of temporalis fascia. The rates of complication, revision surgery, and death were 54%, 26%, and 6%, respectively. Most of the children are now orally fed, and 9 still have a tracheotomy. PMID- 15895788 TI - Frontolateral vertical partial laryngectomy without tracheotomy for invasive squamous cell carcinoma of the true vocal cord: a 25-year experience. AB - On the basis of an inception cohort of 270 patients with a previously untreated invasive squamous cell carcinoma of the true vocal cord (232 T N0M0, 35 T2N0M0, and 3 T3N0M0) and a minimum of 3 years of follow-up, the authors analyze the oncological and functional outcomes following frontolateral vertical partial laryngectomy without tracheotomy. The 5-year Kaplan-Meier actuarial survival estimate ranged from 83.1% for T1 tumors to 67.2% for T2 tumors (p = .005). On univariate analysis, a significant statistical relationship was noted between reduced survival and the following variables: increased age, increased Charlson comorbidity index score over grade 0, increased tobacco intake, increased alcohol intake, increased T stage, local failure, nodal failure, and development of a metachronous second primary cancer. The hospital mortality rate was 0.4%. A significant postoperative surgical complication was noted in 49 patients (18.1%). The predominant significant surgical complication was wound infection (19 patients; 7%), followed by seroma and major subcutaneous emphysema. No significant statistical relationship was noted in a comparison of each, significant postoperative complication (including postoperative death) with the variables under analysis. The incidence of secondary tracheotomy was 0.4%. The incidence of completion laryngectomy due to functional problems was 0%. The 5 year Kaplan-Meier actuarial local control estimate was 91% for T1 tumors and 68.7% for T2 tumors (p <.0001). Within the T1 tumors, the 5-year Kaplan-Meier actuarial local control estimate ranged from 96.2% for tumors without anterior commissure involvement to 74.7% for tumors with anterior commissure involvement (p = .0002). On univariate analysis, a significant statistical relationship was noted between an increase in local recurrence and the following variables: increased T stage, anterior commissure involvement, and pathological margin involvement. The overall disease control rate and laryngeal preservation rate were 92.9% and 93.3%, respectively. PMID- 15895789 TI - Postauricular needle aspiration of subperiosteal abscess in acute mastoiditis. AB - To test the hypothesis that subperiosteal abscess, a complication of acute mastoiditis, can be treated equally well by needle aspiration as by cortical mastoidectomy, we performed a retrospective analysis of 78 pediatric patients hospitalized between 1995 and 2003 and performed an analysis of published data on types and outcomes of treatment approaches for acute mastoiditis. Postauricular pus aspiration resolved the subperiosteal abscess in 14 of 17 patients. The length of the hospital stay of patients who underwent aspiration was shorter than that of patients who underwent cortical mastoidectomy. We conclude that postauricular pus aspiration, a simple and minimally invasive procedure, is an effective treatment modality for subperiosteal abscess. Mastoidectomy should be reserved for nonresponsive cases or those with more serious complications. Broad spectrum antibiotics, myringotomy with daily toilet of the ear, and postauricular aspiration, when required, minimize the indications for surgery and reduce the hospital stay. PMID- 15895790 TI - Hearing loss as a late complication of radiotherapy in children with brain tumors. AB - Late postirradiation hearing loss has been well described in the adult population. Few reports exist on the pediatric population. We conducted a retrospective review of 157 consecutive children with brain tumors treated exclusively with irradiation at St Jude Children's Research Hospital. Twenty-six patients developed a hearing loss, 74 did not, and 57 were excluded because of incomplete records. We report a statistically significant 27.41% cumulative risk of a stringent 20-dB hearing loss in the voice frequency range by the fifth year after radiotherapy. The right side demonstrated a significant frequency effect, with a higher incidence of loss in the higher-frequency region. We found no difference in cumulative incidence of hearing shift between the low-, middle-, and high-frequency ranges for either ear. This risk should be anticipated and managed as part of the treatment plan for radiotherapy for the treatment of malignancies. Radiation-induced hearing loss is important to acknowledge so that techniques of hyperfractionation, total dose, ports, preservative infusion medical therapy, or prolonged medical intervention (such as anticoagulants) can be developed that might reduce this disabling problem of postirradiation sensorineural hearing loss in future patients. PMID- 15895791 TI - Hereditary congenital unilateral deafness: a new disorder? AB - Congenital unilateral deafness is a rare disorder. The prevalence rates are unknown. The prevalence of children with severe to profound hearing losses that are congenital (or acquired before the development of speech and language) is 0.5 to 3 per 1,000 live births. Evidently, congenital unilateral deafness must have a lower prevalence. The purpose of this research was to present a new disorder, hereditary congenital unilateral deafness. A pedigree is presented in which both male and female members display symptoms of congenital unilateral deafness. Two affected persons and a normal-hearing member of the family have vestibular abnormalities without dysequilibrium. The inheritance pattern of this new syndrome is not clear. We hypothesize that the disorder might be new. A family like this has never before been presented in the medical literature. PMID- 15895792 TI - [Epidemiology and prevention of cardiovascular diseases]. AB - Epidemiology and prevention of coronary artery disease. The author summarizes the most important risk factors of atherosclerosis and epidemiological data of coronary heart disease. According to epidemiological data during the next 10-20 years cardiovascular diseases will be the leading cause of death all around the world. Smoking is the most important risk factor causing half of all avoidable death, half of these deaths caused by cardiovascular diseases. Three strategies of prevention are discussed: population strategies, high risk strategy and secondary prevention. All of these preventive strategies should be implemented to reach the goal. During the last years many drug studies were finished proving the effectiveness of aspirin, statins, ACE inhibitors and beta blockers as an effective tool of prevention. PMID- 15895793 TI - [Roma colonies in Hungary--medical care of children and hygienic conditions]. AB - INTRODUCTION: The hygienic conditions and the public health safety of the Hungarian Roma living in colonies are poorly known, the health care of Romany children is often subject to criticism. AIM: The authors aim was to identify the blank areas present in public health and to analyze the medical care of children living in Gipsy colonies. METHODS: Medical care of children living in Gipsy colonies in Hungary and the hygienic situation of these colonies have been analyzed. The examinations, performed by the staff of the National Public Health and Medical Officers' Service included all counties and the capital. Data were gathered from answers given to the questions in the circular sent by the Chief Medical Officer of State, by questionnaire-interview method as well as, by on site hygienic examination. Data from the series of examinations were completed by the data of the National Institute for Basic Medical Care. RESULTS: According to the hygienic definition of Romany colonies, there are 767 colonies in Hungary, scattered in 530 habitats, while according to the definition of availability of public utilities and structure of the colonies the number of colonies is 300-400. There are no Romany colonies in the capital, while most of them are located in the counties Hajdu-Bihar, Baranya, Szabolcs-Szatmar-Bereg and Borsod-Abauj Zemplen. A population of 3 million is living in the habitats with Romany colonies; 138000 (4,7%) of them are living in Romany colonies, or in colony-like communities with similar hygienic situation. In these habitats 5,8% and 16,3% of the population are children of 0-5 years old and 0-14 years old, respectively, while in the colonies the same numbers are 15,8% and 35,6%. Medical care is provided by 456 family pediatrist for the the habitats with Romany colonies. Consulting hours for the children population of 11,4% of the family doctors seems to be insufficient (< 15 hours/day/5 days/week). Only 60% of care for children is provided by specialist in pediatrics. 94% of emergency care for children is provided for family doctors of the adult population (this proportion is characteristic for the whole country). 12,5% of district nurses caring for the colonies provide care for more than 2 habitats; 147 of them (22% of all district nurses) provide care for more than 750 persons (experiences show that appropriate care can be ensured if the number of care recipients does not exceed 300 and the maximum number of settlements under care is two). Illegal waste deposits and animal carcase disposal site are located within 1000 m of 15% and 11% of the colonies, respectively. Most of the dwelling sites are messy, hygienically neglected, rodents and unvaccinated stray dogs are frequently met in their neighborhood. CONCLUSIONS: 1. sufficiently accurate data are not available on the Gipsy colonies (number, location, population, children population), partly due to the related different definitions and partly because this issue of public health importance is not treated uniformly. The repeated examinations of the National Public Health and Medical Officers' Service presented here only improved the relevant information; 2. child health care of the Gipsy colonies does not differ from that of the rest of the Hungarian children, but similarly to that, a lot of corrections are needed--especially regarding the emergency service; 3. the hygienic situation of the Gipsy colonies is not acceptable, it endangers the public health and epidemiological safety of not only their inhabitants, but the whole country. PMID- 15895794 TI - [Diagnostic difficulties in kidney cancer filling up the retroperitoneal space]. AB - An autopsy case of a huge, rapidly progressive Bellini duct carcinoma is presented. During a 6-month period the patient was hospitalized many times in different medical units but her illness has not been diagnosed until the final weeks. The author discusses the pitfalls of diagnosis from clinicopathological point of view. PMID- 15895795 TI - [The state and models of specialisation in geriatrics in Europe and Hungary]. AB - This article is based on a lecture held in the 1st Debrecen Days of Internal Medicine, in which the author summarizes the situation of specialisation in geriatrics in EU members and Hungary. There are significant differences between countries in this field. Not all EU member countries accept geriatrics as an independent specialty, but the education in graduate level is present uniquely. Geriatrics is accepted as a specialty from 2000 in Hungary and there are nearly 100 geriaters up till now. Higher and higher percentage of european population is in age over 65 years, so the importance of education of geriatrics is a great demand in the near future. PMID- 15895796 TI - [Untreated hyperthyroidism with severe cardiac complication in adolescence--a case study]. AB - A 16-years old boy with poor family circumstances was admitted to hospital due to severe cardiac failure (stagnation in the pulmonary and systemic circulations, ejection fraction: 29%, pericardial fluid, atrial flatter) needing intensive care. Diagnostic tests revealed extreme hyper-function of the thyroid as the underlying cause. After stabilising the cardiac and hormonal states of the patient with medication, subtotal resection of the thyroid was carried out since the poor family circumstances did not make the provision of long-term thyreostatic pharmacotherapy feasible. Cardioversion solved the arrhythmia. The patient was discharged from hospital with stable circulation and normal thyroid functions after 2 months. The authors report the very uncommon complications of an endocrine syndrome with autoimmune origin rarely observed in childhood. They highlight the importance of early diagnosis and the immediate start of adequate therapy for good long-term outcome. PMID- 15895797 TI - [Distribution of haplotypes for four Y-sTR loci and validation in forensic science by using a double-fluorescent multiplex PCR system]. AB - OBJECTIVE: We focus on developing a multiplex PCR system for Y-STR loci that can be detected by double fluorescent system and assessing their usefulness in forensic mixture samples. METHODS: The primers of four Y-STR loci (DYS-GATA-A10, DYS531, DYS557 and DYS448) amplified by multiplex PCR technique were labeled with fluorescence, then the PCR products of these Y-STRs loci were detecting and typing by ABI PRISM310 Genetic Analyzer. RESULTS: When 120 unrelated individuals from the Han population in Chengdu were detected by the system, Y-GATA-A10, DYS531, DYS557 and DYS448 showed 5, 5, 8, 7 alleles, respectively. A total of 78 different haplotypes was identified and the genetic diversity reached 0.9881. To the three cases of mixture stains failed by using conventional autosomal STR analysis, our multiplex system drew conforming conclusion comparing to the suspect's Y-STRs genotypes. CONCLUSION: Our results show that the multiplex system of four Y-STR will be very powerful for Y-STR database establishing, the paternity testing and mixture stains identifying. PMID- 15895798 TI - [DNA samples preparation from single cell and its application in sensitivity test]. AB - OBJECTIVE: To establish a reliable, exact and practical method to prepare DNA samples for sensitivity-test purposes. METHODS: The micromanipulation method was employed to prepare exact quantity DNA samples used to study the sensitivity of Profiler Plus Kit-ABI PRISM 310 system. RESULTS: We succeed in establishing a micromanipulation method to prepare groups of DNA samples, which contain 1-11 cells in turn, and also succeed in using them to study the sensitivity of Profiler Plus Kit-ABI310 system. CONCLUSION: The method we have established is proved to be a reliable, exact and practical way to prepare DNA samples for sensitivity-test purposes. PMID- 15895799 TI - [Detecting haplotypes of four Y-STR by multiplex amplification]. AB - OBJECTIVE: To establish a multiplexing Y-STR system and study haplotype frequencies of 4 Y-specific loci in China Han population. METHODS: DYS439, DYS390, GATA-A7.2 and DYS393 loci were amplified simultaneously and were analyzed by polyacrylamide gel electrophoresis and silver staining. RESULTS: When 558 unrelated male individuals from the Han population in China were tested by the multiplex system, DYS439, DYS390, GATA-A7.2 and DYS393 show 7,7,7,6 alleles, respectively. 180 different haplotypes were detected. The power of discrimination of this system was 0.9853. CONCLUSION: The multiplex amplified system of these 4 Y-specific loci and their database are useful for human origin exploration and forensic practice. PMID- 15895800 TI - [A simple and rapid modified--new method for SNP typing by fragment length discrepant allele specific PCR]. AB - OBJECTIVE: To establish a new method for single nucleotide polymorphism (SNP) typing based on allele specific PCR: fragment length discrepant allele specific PCR (FLDAS-PCR), and study the influence on specific extension by introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers. METHODS: For SNP loci rs759117 and rs760887, two allele specific forward primers, with different length and a mismatch introduced at the third or fourth 3' terminal base, and a public reverse primer were designed for SNP typing. The genotyping of SNP was determined by the two allele specific fragments different in size after polyacrylamide gel and silver staining. RESULTS: The different homozygote genotypes comprised a single band with different size respectively, and the heterozygote genotypes comprised two bands. Typing results were completely consistent with those by direct sequencing. Non-specific primer extension was decreased remarkably after introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers, and the stringency of PCR reaction was cut down. CONCLUSION: FLDAS-PCR is a simple, rapid and efficient new method for SNP typing. During FLDAS-PCR, specific primers with a mismatch at the third or fourth 3'-terminal base have more power to identify two alleles. PMID- 15895801 TI - [Comprehensively analysis the correlation between the height of a person and the length of his/her footprint]. AB - OBJECTIVE: Comprehensively analysis the correlation and the law of variation between the height of a person and the length of his/her footprint. METHODS: Collecting footprint samples of those people of different age and sex from different area all of our country, and adopt the regressive analysis method to study these samples. RESULTS: A sum of useful data and regressive equation were obtained. CONCLUSION: The correlation between the height of a person and the length of his/her footprint is obviously, the approximate height of a person can be reckoned according to thelength of his/her footprints. PMID- 15895802 TI - [A preliminary study on estimation of postmortem interval according to beta-actin mRNA stability in rat]. AB - OBJECTIVE: In order to find a new parameter to estimate the postmortem interval, beta-actin mRNA in lung and thoracic muscle of rats was detected at different time point postmortem. METHODS: Rats were killed by neck dislocation and left in a temperature controlling system at 21 degrees C for 12 days postmortem. Total RNA in lung and thoracic muscle at different time point was extracted and beta actin mRNA expression was examined by RT-PCR. Semi-quantification analysis of the image of electrophoresis was performed to confirm the changes of beta-actin mRNA expression. RESULTS: beta-actin mRNA in lung of rats still could be detected at 12 days postmortem, but-disappeared in thoracic muscle at 8 days postmortem. CONCLUSION: The expression of beta-actin mRNA in lung and thoracic muscle could be a new parameter for estimation of PMI. PMID- 15895803 TI - [Primarily study and significance of ET-1 during ischemia with RT-PCR]. AB - OBJECTIVE: To explore the relationship between expression of endothelin in heart and ischemia. METHODS: Using the RT-PCR method, we enquiry changes of ET-1 RNA after 60min ischemia in hearts of rats. RESULTS: Although the two group appear positive, they are different significantly, which indicate that ischemia may lead to the increase of ET-1 mRNA. CONCLUSION: The experiment, therefore, pave a way for immunochemical study. PMID- 15895804 TI - [A study on the expression of HSP70 and iNOS after human brain contusion]. AB - OBJECTIVE: To study the relationship between expression of HSP70, iNOS and traumatic brain contusion (TBI) in different posttraumatic intervals. METHODS: 35 samples of brain contusion were examined using immunohistochemecal staining to evaluate the expression of HSP70 and iNOS. RESULTS: Maximal HSP70 expression was found at 0h after brain contusion. The intensity of HSP70 staining decreased remarkably to the minimum at 24h after TBI, then increased gradually. Expression of iNOS positive cells increased significantly and reached the maximum level 48h after TBI, then the expression decreased gradually from the 2nd day to the 11th day. CONCLUSION: The changes of HSP70 and iNOS immunohistochemical staining can be used as a referential data for estimating time interval after human brain contusion. PMID- 15895805 TI - [Detecting DNA damage of cell in rats using comet assay after tetramine poisoned]. AB - OBJECTIVE: To study the damage of DNA in lymphocytes, brain cells and cardiac muscle cells of rats induced by different dose of tetramine and to speculate the toxicant mechanism of tetramine. METHODS: The rat were poisoned by Tetramine, which was taken in by mouth. The rat poisoning models were used by 0.2, 0.1, 0.05, 0.01 mg x kg(-1) Tetramine, and comparison model was made by NS. Lymphocytes and brain cells and cardiac muscle cells of rats were separatd and collected form experimentation rat. DNA damages of cells which were exposed to different doses of tetramine were detected using the single cell gel electrophorresis (SCGE) or comet assay. RESULTS: DNA damages have been observed in lymphocytes, brain cells and cardiac muscle cells of rats which exposed form 0.01mg x kg(-1) doses of tetramine to 0.2mg x kg(-1) doses of tetramine. The test groups are very significantly statistical different to the control group (P<0.01). CONCLUSION: It is assumed that DNA damages of cells might be one of the toxicant mechanism of tetramine. PMID- 15895806 TI - [The diagnostic value of SEP in LDH and its forensic significance]. AB - OBJECTIVE: To evaluate the diagnostic value of somatosensory evoked potential(SEP) in lumbosacral disc herniation(LDH). METHODS: Posterior tibial nerve somatosensory evoked potential(PTNSEP) and dermatomal somatosensory evoked potential (DSEP) were recorded from 60 patients with radiculopathy caused by LDH and 30 healthy subjects. RESULTS: DSEP was abnormal in 56 cases (93.3%) and PTNSEP was abnormal in 26 cases (43.3%). The significant difference was observed (P<0.001). The abnormality of DSEP from L4 dermatome was detected mainly in patients with L3-4 LDH, the abnormality of DSEP from L5 dermatome was detected mainly in patients with L4-5 LDH, the abnormality of DSEP from L5S1 dermatome was detected mainly in patients with S1 LDH. DSEP can reflect the compressed extent of nerve root. CONCLUSIONS: There was a good correlation of DSEP with lumbosacral nerve root injury,which supplement the information that can not be provided by imaging examingation. DSEP is a sensitive electrophysic method that not only is used to evaluate single nerve root function, but also is helpful in the location of the lesion. PMID- 15895807 TI - [Separation and determination of aconitine in medicine wine by reversed-phase high performance liquid chromatography]. AB - OBJECTIVE: For the purpose of more accurate and rapid analysis of aconitine in medicine wine, a reversed-phase High Performance Liquid Chromatography (HPLC) method was developed. METHODS: Standard aconitine was added to the blank wine when the wine sample was pretreated. The pretreatment method of samples, the linear range, the precision, the recoveries in the plasma were investigated by using of white wine plasma spiked with standard aconitine. RESULTS: The linear range was 0.45 approximately 9.0 microg x mL(-1), r=0.998 8. The detective limit was 0.45 microg x mL(-1), The intra and inter-day precision of assay for aconitine were less than 3.1% and 4.7% (n=5) in wine respectively. The recoveries of aconitine were more (97.3+/-2.8)% in medicine wine. The HPLC method has been used to investigate the concentration of aconitine in one forensic medicine case. CONCLUSION: The HPLC method of quantitative analysis aconitine is rapid and sensitive. It is only in the 2.0 h that determination of aconitine in the medicine wine. PMID- 15895808 TI - [Concentration and determination of strychnine alkaloid in biological fluids]. AB - OBJECTIVE: To establish a new method for determination of strychnine alkaloid in biological fluids based on molecularly imprinted polymers. METHODS: A strychnine molecularly imprinted monolithic column was prepared by in-situ molecularly imprinted technique. The polymer was filled to a 1cm column, and a method was developed to concentrate and determine strychnine alkaloids in biological fluids. RESULTS: the limit of detection of the method was 4.9 ng, and the recoveries were more than 92%. The relative standard deviations were smaller than 6.59%. The linear correlation coefficients of standard curves were 0.999 1 and 0.9966 respectively. This method was applied to concentrate and determine strychnine in plasma and urine of poisoned rabbit. CONCLUSION: The new method could concentrate and simultaneously determine strychnine alkaloids in biological fluids, and it was applied to forensic toxicological analysis. PMID- 15895809 TI - [Advances in the research of vitamin D receptor gene]. AB - The polymorphism of vitamin D receptor (VDR) gene may be associated with bone density, osteoporosis and body height. In this review, the authors summarize the association of the four VDR single nucleotide polymorphisms (SNPs) loci FokI, BsmI, ApaI, TaqI with bone density, osteoporosis and body height reported at home and abroad in recent years. And find that though each country's report may be different, its valuable in the field of genetics, clinical endocrinology and metabolism, especially in the research area of body height in forensic medicine. PMID- 15895810 TI - [The research progress in Marfan syndrome]. AB - Marfan syndrome (MFS) is a potentially fatal connective disorder that is inherited as an autosomal dominant trait with a prevalence of around 2-3 in 10000 live births. It is characterized by defects in the cardiovascular, skeletal and ocular systems. Evidence from genetic indicates that mutations in FBN1, the gene that encodes fibrillin-1 are responsible for MFS. In addition to skeletal, ocular, and cardiovascular feathers, patients with MFS have also involvement of skin, integument, lungs, and muscle tissue, and the condition in sudden death is also very common due to severe abnormalities of cardiovascular system. PMID- 15895811 TI - [Applications of DNA methylation markers in forensic medicine]. AB - DNA methylation is a post-replication modification that is predominantly found in cytosines of the dinucleotide sequence CpG. Epigenetic information is stored in the distribution of the modified base 5-methylcytosine. DNA methylation profiles represent a more chemically and biologically stable source of molecular diagnostic information than RNA or most proteins. Recent advances attest to the great promise of DNA methylation markers as powerful future tools in the clinic. In the past decade, DNA methylation analysis has been revolutionized by two technological advances--bisulphite modification of DNA and methylation-specific polymerase chain reaction (MSP). The methylation pattern of human genome is space time specific, sex-specific, parent-of-origin specific and disease specific, providing us an alternative way to solve forensic problems. PMID- 15895812 TI - [Molecular biology advance in the Rh blood group system]. AB - The Rh blood group system is one of the most complex of the known human blood group polymorphisms, including above 45 blood group antigens. Considerable progress has been made in our understanding of the molecular basis of Rh in the past 10 years. The bases of Rh inheritance, RH gene and its evolution, the structure and function of Rh complex, as well as nonerythroid Rh homolog, have been determined. Further improvements have been made in the technology of Rh genotyping. The review provides an update on the advance of Rh blood group to give information in the practice of forensic science. PMID- 15895813 TI - [The study of the sequence of molecular markers of mitochondrial DNA of Sarcosaphagous Flies]. AB - Identifying sarcosaphagous flies specimens is an important first step in a forensic-entomological analysis. It is traditionally performed using morphological features of the Sarcosaphagous Flies. However, Morphological identification may be complicated by the numerical diversity of species and physical similarity between different species, particularly in immature stages. The sequences focused on some sections of the cytochrome oxidase I and II (CO I and CO II) encoding region of mtDNA could be as the prospective basis of a diagnostic technique. By Analysis of these sequences, forensic doctors can reveal abundant phylogenetic informative nucleotide substitutions that could effectively identify Sarcosaphagous flies to species group. It was not reported in our country before and was reviewed in this article now. PMID- 15895814 TI - Face value: challenges of transplant technology. PMID- 15895815 TI - Ethos and economics: examining the rationale underlying stem cell and cloning research policies in the United States, Germany, and Japan. PMID- 15895816 TI - Ethical and legal aspects of using an identical twin as a skin transplant donor for a severely burned minor. PMID- 15895817 TI - Defining a standard of care in the practice of acupuncture. PMID- 15895818 TI - Phorbol ester differentiates the levels of [3H]MK-801 binding in rats lines selected for differential sensitivity to the hypnotic effects of ethanol. AB - These studies addressed the possible involvement between sensitivity to the hypnotic action of ethanol and function of the NMDA receptor. The studies were carried out using high-alcohol sensitive (HAS) and low-alcohol sensitive (LAS) rats, two rats having differential sensitivity to the acute hypnotic action of ethanol. The animal models were developed by a selective breeding experiment. Using a quantitative autoradiograph technique, it was demonstrated that [3H]MK 801 binding to the NMDA receptor was highest in hippocampus in both HAS and LAS rats, but significant [3H]MK-801 binding was also detected in cortex, caudate putamen, and thalamus of HAS and LAS rats. The density of [3H]MK-801 binding was lower only in cerebellar granule layers of untreated HAS rats as compared to the same brain area in untreated LAS rats. Activation of protein kinase C (PKC) by 100 nM PDBu, increased [3H]MK-801 binding in cortex, caudate-putamen, thalamus, central gray, and cerebellum of HAS rats but activation of PKC did not influence [3H]MK-801 binding in LAS rats. These activation of PKC differentiates between [3H]MK-801 binding of HAS and LAS rats in frontal cortex (layer II-IV and cingulate), caudate-putamen, and ventral lateral thalamic nuclei. The basal level of PKC-gamma mRNA was higher in HAS rats than that of LAS rats. These results suggest that the activation of PKC potentiates NMDA receptor function of the rat line which is more sensitive to alcohol (HAS) but does not affect [3H]MK-801 binding of alcohol resistant (LAS) rats. PMID- 15895819 TI - A proton-dependent zinc uptake in PC12 cells. AB - Intracellular pH in pheochromocytoma (PC12) cells was manipulated by 'acid loading' the cells and the effect of such a change on radioactive zinc uptake was studied. It was found that zinc uptake was stimulated in cells loaded with protons without causing any measurable change in the intracellular pH. To confirm our assumption that the proton flux due to zinc entry is too small to be measured, we calculated the pH change that one would expect because of zinc influx. The intrinsic buffer capacity of PC12 cells was determined to be 8.03 mM/pH unit and was used in these calculations. It was found that at the five minute incubation, zinc uptake occurring under our experimental conditions could cause a pH change of 0.000277 pH units per minute (assuming a 1:2 zinc:proton stoichiometry). This study adds a new dimension towards understanding the role played by intracellular pH in causing zinc entry into cells. PMID- 15895820 TI - 2,5-Hexanedione induced decrease in cytoskeletal proteins of rat sciatic-tibial nerve. AB - Exposure chronically to n-hexane produces peripheral-central axonopathy mediated by 2,5-hexanedione (HD). Previous studies have demonstrated decreases in neurofilament (NF) contents of peripheral and central nervous regions from rats intoxicated with HD, and recent analysis has demonstrated that axonal atrophy, instead of NF-filled swellings, is a specific component of morphologic alterations. To deeply investigate the alterations of cytoskeletal proteins in HD peripheral neuropathy, the relative levels of NF-L, NF-M, NF-H, alpha-tubulin, beta-tubulin and beta-actin of rat sciatic-tibial nerves were determined by SDS PAGE and immunoblotting. HD was administrated to Wistar rats by intraperitoneal injection at dosage of 200 or 400 mg/kg/day (five-times per week). Rats were sacrificed after 6 weeks of treatment, and sciatic-tibial nerves were dissected, homogenized, and used for the determination of cytoskeletal proteins. Except for supernatant NF-L that could not be assayed, the results showed HD intoxication was associated with significant decreases in NF subunits in both of the supernatant and the pellet fractions of sciatic-tibial nerve homogenates (P < 0.01), and obvious reductions in alpha-tubulin, beta-tubulin and beta-actin only in the supernatant (P < 0.05 or P < 0.01). Among these alterations, the falls in the levels of NF subunits tended to be greater compared to those of the other cytoskeletal proteins in all HD-exposed groups, and the trend for decrements in NF-M was greater than those in the other NF subunits. Thus, HD intoxication was associated with significant declines in cytoskeletal protein contents in rat sciatic-tibial nerves, and the decreases might be related to the involvement of the peripheral axonopathy induced by HD. PMID- 15895821 TI - Short- and long-term memory are differentialy modulated by hippocampal nerve growth factor and fibroblast growth factor. AB - Rats were implanted with cannulae in the CA1 area of the dorsal hippocampus and trained in one-trial step-down inhibitory avoidance. Two retention tests were carried out in each animal, one at 1.5 h to measure short-term memory (STM) and another at 24 h to measure long-term memory (LTM). The purpose of the present study was to evaluate the modulation on hippocampal nerve growth factor (NGF) and basic fibroblast growth factor (bFGF) on short- and long-term memory. Immediately after training, animals received 5 microl of NGF (0.05, 0.5 or 5.0 ng), bFGF (1.25, 12.5 or 125 ng) or saline per side. At the higher dose, NGF blocked STM. In contrast, NGF at dose of 0.5 and 5.0 ng improved LTM. The bFGF infusion at a dose of 125 ng enhanced LTM. However, bFGF did not alter STM. These findings indicate that hippocampal NGF and bFGF modulate STM and LTM in a different manner. PMID- 15895822 TI - Estradiol protects against oxygen and glucose deprivation in rat hippocampal organotypic cultures and activates Akt and inactivates GSK-3beta. AB - Here we investigated the neuroprotective effect of 17beta-estradiol in an in vitro model of ischemia. We used organotypic hippocampal slice cultures, acute or chronically treated with 17beta-estradiol (10 nM), and exposed to oxygen and glucose deprivation (OGD). Cellular death was quantified by measuring uptake of propidium iodide (PI), a marker of dead cells. In OGD exposed cultures, treated only with vehicle, about 70% of the CA1 area of hippocampus was labeled with PI, indicating a great percentage of cellular death. When cultures were treated with 17beta-estradiol (acute or chronically), this cellular death was reduced to 15%. This effect was prevented by LY294002 but was not by PD98059. Immunoblotting revealed that both, chronic and acute, treatments with 17beta-estradiol induced the phosphorylation/activation of Akt and the phosphorylation/inactivation of GSK 3beta. Our results show a clear neuroprotective effect of 17beta-estradiol and suggest that this effect could involve PI3-K pathway. PMID- 15895823 TI - Histological co-localization of iron in Abeta plaques of PS/APP transgenic mice. AB - This study confirms the presence of iron, co-localized with Abeta plaques, in PS/APP mouse brain, using Perls' stain for Fe3+ supplemented by 3,3' diaminobenzidine (DAB) and Abeta immunohistochemistry in histological brains sections fixed with formalin or methacarn. In this study, the fixation process and the slice thickness did not interfere with the Perls' technique. The presence of iron in beta-amyloid plaques in PS/APP transgenic mice, a model of Alzheimer's disease (AD) pathology, may explain previous reports of reductions of transverse relaxation time (T2) in MRI studies and represent the source of the intrinsic Abeta plaque MR contrast in this model. PMID- 15895824 TI - Pattern of glial fibrillary acidic protein expression following kainate-induced cerebellar lesion in rats. AB - In the present study glial fibrillary acidic protein (GFAP) expression was assessed following intravermian injection of kainic acid (KA) or physiological saline to adult rat cerebellum. After 2- to 30-day recovery period, free-floating sections cut with a microtome were obtained and were proccessed for immunocytochemistry against GFAP. Injection of both kainate and physiological saline elicited significant astrogliotic reaction, i.e. in the area around the lesion thick GFAP-positive Bergmann fibers with typical orientation appeared in the molecular and hypertrophied astrocytes abundantly appeared in the granular layer. However, following kainate intoxication lesion was not surrounded by typical demarcation glial scar during 30-day recovery period in contrast to the appearance of usual glial scar in the group injected with physiological saline, as early as 7-day postlesion. Preserved spatial organization of Bergmann fibers and the absence of typical demarcating glial scar after kainate-induced cerebellar lesion suggest distinct pattern of astrogliosis that presents an interesting model system to study the importance of glial scar in the recovery after ischemic brain insults. PMID- 15895825 TI - Effects of L-mono methyl-arginine, N-acetyl-cysteine and diphenyleniodonium on free radical release in C6 glial cells enriched in hexacosenoic acid. AB - Previously, we have shown that C6 glial cells enriched in hexacosenoic acid (HA) incubated with oxidative stressors released higher amounts of nitric oxide (NO) products and superoxide (O2(-)), compared to native C6 cells. In the present study, we examined the effects of pretreatment with some of free radical release inhibitors. The aim was to determine the origin of the enhanced generation of NO and superoxide, and to test the possibility of preventing it. Pre-treatment with L-mono-methyl-arginine and N-acetyl-cysteine in oxidized low-density lipoprotein (ox-LDL) exposed HA cells, inhibited not only nitrite but also superoxide production suggesting that O2(-) anion could partially derive from inducible NO synthase. We also observed that ox-LDL treatment of HA cells reduced the intracellular glutathione levels and activated extracellular signal-related kinases. Since this signalling is related to neurotoxic effect, our data substantiate the role of the free radicals in X-linked adrenoleukodystrophy pathogenesis, as HA cells have been used as an in vitro model for this disease. PMID- 15895826 TI - Krebs cycle intermediates modulate thiobarbituric acid reactive species (TBARS) production in rat brain in vitro. AB - The aim of this study was to investigate the effect of Krebs cycle intermediates on basal and quinolinic acid (QA)- or iron-induced TBARS production in brain membranes. Oxaloacetate, citrate, succinate and malate reduced significantly the basal and QA-induced TBARS production. The potency for basal TBARS inhibition was in the order (IC50 is given in parenthesis as mM) citrate (0.37) > oxaloacetate (1.33) = succinate (1.91) > > malate (12.74). alpha-Ketoglutarate caused an increase in TBARS production without modifying the QA-induced TBARS production. Cyanide (CN-) did not modify the basal or QA-induced TBARS production; however, CN- abolished the antioxidant effects of succinate. QA-induced TBARS production was enhanced by iron ions, and abolished by desferrioxamine (DFO). The intermediates used in this study, except for alpha-ketoglutarate, prevented iron induced TBARS production. Oxaloacetate, citrate, alpha-ketoglutarate and malate, but no succinate and QA, exhibited significantly iron-chelating properties. Only alpha-ketoglutarate and oxaloacetate protected against hydrogen peroxide-induced deoxyribose degradation, while succinate and malate showed a modest effect against Fe2+/H2O2-induced deoxyribose degradation. Using heat-treated preparations citrate, malate and oxaloacetate protected against basal or QA induced TBARS production, whereas alpha-ketoglutarate induced TBARS production. Succinate did not offer protection against basal or QA-induced TBARS production. These results suggest that oxaloacetate, malate, succinate, and citrate are effective antioxidants against basal and iron or QA-induced TBARS production, while alpha-ketoglutarate stimulates TBARS production. The mechanism through which Krebs cycle intermediates offer protection against TBARS production is distinct depending on the intermediate used. Thus, under pathological conditions such as ischemia, where citrate concentrations vary it can assume an important role as a modulator of oxidative stress associated with such situations. PMID- 15895827 TI - Glutamate regulates dystrophin-71 levels in glia cells. AB - Glial glutamate receptors are likely to be involved in neuronal differentiation, migration, and plasticity. Dystrophin, the protein defective in Duchenne muscular dystrophy (DMD) is widely expressed in the Central Nervous System. Activation of internal promoters of the DMD gene leads to the production of several proteins, the Dystrophin-71 (Dp-71) being the most abundant in the encephalon. This protein is known to stabilize neurotransmitter receptors in clusters and its absence has been correlated with cognitive deficits in a mouse model. Using cultured chick Bergmann glia cells and mouse cerebellar fusiform astrocytes, we demonstrate here that glutamate receptor activation results in a time and dose dependent decrease of Dp-71 levels. This effect is mediated through alphaamino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptors. The present results suggest an involvement of Dp-71 in glutamate receptor signaling and possibly clustering and further support the notion of an active role of glia in the physiology of glutamatergic transmission. PMID- 15895828 TI - Cyclosporin-A inhibits constitutive nitric oxide synthase activity and neuronal and endothelial nitric oxide synthase expressions after spinal cord injury in rats. AB - Nitric oxide (NO) plays a role in the pathophysiology of spinal cord injury (SCI). NO is produced by three types of nitric oxide synthase (NOS) enzymes: The constitutive Ca2+/calmodulin-dependent neuronal NOS (nNOS) and endothelial NOS (eNOS) isoforms, and the inducible calcium-independent isoform (iNOS). During the early stages of SCI, nNOS and eNOS produce significant amounts of NO, therefore, the regulation of their activity and expression may participate in the damage after SCI. In the present study, we used Cyclosporin-A (CsA) to further substantiate the role of Ca-dependent NOS in neural responses associated to SCI. Female Wistar rats were subjected to SCI by contusion, and killed 4 h after lesion. Results showed an increase in the activity of constitutive NOS (cNOS) after lesion, inhibited by CsA (2.5 mg/kg i.p.). Western blot assays showed an increased expression of both nNOS and eNOS after trauma, also antagonized by CsA administration. PMID- 15895829 TI - Exposure of postnatal rats to a static magnetic field of 0.14 T influences functional laterality of the hippocampal high-affinity choline uptake system in adulthood; in vitro test with magnetic nanoparticles. AB - Our previous experiments indicated an age- and sex-dependent functional lateralization of a high-affinity choline uptake system in hippocampi of Wistar rats. The system is connected with acetylcholine synthesis and also plays a role in spatial navigation. The current study demonstrates that a single in vivo exposure of 7- or 14-day-old males to a static magnetic field of 0.14 T for 60 120 min evokes asymmetric alterations in the activity of carriers in adulthood. Namely, the negative field (antiparallel orientation with a vertical component of the geomagnetic field) mediated a more marked decrease in the right hippocampus. The positive field (parallel orientation) was ineffective. Moreover, differences between the carriers from the right and the left hippocampi were observed on synaptosomes pretreated with superparamagnetic nanoparticles and exposed for 30 min in vitro. The positive field enhanced more markedly the activity of carriers from the right hippocampus, the negative that from the left hippocampus, on the contrary. Our results demonstrate functionally teratogenic risks of the alterations in the orientation of the strong static magnetic field for postnatal brain development and suggest functional specialization of both hippocampi in rats. Choline carriers could be involved as secondary receptors in magnetoreception through direct effects of geomagnetic field on intracellular magnetite crystals and nanoparticles applied in vivo should be a useful tool to evaluate magnetoreception in future research. PMID- 15895830 TI - Role of ERK in hydrogen peroxide-induced cell death of human glioma cells. AB - Oxidative stress is known to induce cell death in a wide variety of cell types, apparently by modulating intracellular signaling pathways. Activation of extracellular signal-regulated kinase (ERK) in oxidative stress remains controversial. In some cellular systems, the ERK activation is associated with protection against oxidative stress, while in other system, the ERK activation is involved in apoptotic cell death. The present study was undertaken to examine the role of ERK activation in H2O2-induced cell death of human glioma (A172) cells. H2O2 resulted in a time- and dose-dependent cell death, which was largely attributed to apoptosis. H2O2 treatment caused marked sustained activation of ERK. The ERK activation and cell death induced by H2O2 was prevented by catalase, the hydrogen peroxide scavenger, and U0126, an inhibitor of ERK upstream kinase MEK1/2. Transient transfection with constitutive active MEK1, an upstream activator of ERK1/2, increased H2O2-induced cell death, whereas transfection with dominant-negative mutants of MEK1 decreased the cell death. The ERK activation and cell death caused by H2O2 was inhibited by antioxidants (N-acetylcysteine and trolox), Ras inhibitor, and suramin. H2O2 produced depolarization of mitochondrial membrane potential and its effect was prevented by catalase and U0126. Taken together, these findings suggest that growth factor receptor/Ras/MEK/ERK signaling pathway plays an active role in mediating H2O2 induced apoptosis of human glioma cells and functions upstream of mitochondria dependent pathway to initiate the apoptotic signal. PMID- 15895831 TI - Tra2alpha promotes RA induced neural differentiation of P19 cells. AB - The present study demonstrates a high level of the nuclear Transformer 2alpha (Tra2alpha) protein in adult mouse brain relative to other tissues, including muscle, heart, liver, lungs, kidney and small intestine, suggesting the potential importance of Tra2alpha in neural function. The level of Tra2alpha in mouse cerebrum is developmentally regulated, peaking at neonate stage. In P19 carcinoma cells, Tra2alpha is transiently up-regulated upon retinoic acid (RA) treatment. Although over-expression of Tra2alpha protein alone does not elicit P19 differentiation, under these conditions the response of P19 cells to RA is significantly increased. The results suggest that Tra2alpha proteins may act as a mediator in the signal pathway associated with RA-induced differentiation in P19 cells. PMID- 15895832 TI - Phosphothreonine-212 of Alzheimer abnormally hyperphosphorylated tau is a preferred substrate of protein phosphatase-1. AB - We isolated and characterized several phosphoseryl/phosphothreonyl phosphatase activities (P1-P11) from frontal lobe of six autopsied human brains. Of these, PP1 (P3) was a major tau phosphatase. The enzyme required metal ions and was maximally activated by Mn2+. Western blots with antibodies to known protein phosphatases showed PP1 and PP2B immunoreactivity. However, the removal of PP2B by immunoabsorption or its inhibition with EGTA did not result in appreciable loss of P3 activity. These observations suggest that P3 was an enriched PPI. Dephosphorylation of Alzheimer disease hyperphosphorylated tau (AD P-tau) by PP1 was site-specific. PPI preferentially dephosphorylated pT212 (40%), pT217 (26%), pS262 (33%), pS396 (42%) and pS422 (31%) of AD P-tau. Dephosphorylation of tau at pT181, pS199, pS202, pT205, pS214, and pS404, was undetectable. Of the sites dephosphorylated, pT212 was only a substrate for PP1, as purified/enriched PP2A and PP2B from the same brains did not dephosphorylate this site. PMID- 15895833 TI - Pain effects of glutamate injections into human jaw or neck muscles. AB - AIMS: To document and compare the intensity, localization, and quality of pain evoked by glutamate injections into the human masseter or splenius muscles and to determine the effect of glutamate-evoked pain on the pressure pain thresholds (PPTs) in both jaw and neck muscles. METHODS: Twenty-six healthy men were given painful injections of glutamate (1.0 mol/L) and control injections of isotonic saline (0.165 mol/L) into the masseter and splenius muscles. The subjects rated the perceived intensity of pain on a visual analog scale (VAS), drew the area of the pain on maps of the face and neck, and filled out a Danish version of the McGill Pain Questionnaire (MPQ). PPTs were used to assess the sensitivity of the masseter and splenius muscles to mechanical stimuli (n = 11). RESULTS: Glutamate injection into the masseter or splenius evoked pain lasting almost 10 minutes. Peak pain intensity usually occurred within 2 minutes of the injection, and VAS scores of peak pain were significantly higher for the masseter muscle compared with the splenius muscle (paired t test, P = .003). The pain area from the masseter injections did not extend into the neck region, although in some subjects the pain from the neck region extended into the temporal region. There were no significant relationships between the area of perceived pain and the VAS pain scores (Pearson correlation, P > .297). Glutamate-evoked pain in either the masseter or splenius muscles was associated with significant decreases in masseter or splenius PPTs, respectively (2-wayANOVAs, P < .016). Isometric saline injections were almost pain-free and caused no PPT changes. CONCLUSION: The data suggest that the masseter muscle is more sensitive to glutamate injections and mechanical stimuli than the splenius muscle. The relatively limited overlap between the sensory manifestations of pain from masseter and splenius muscles may have potential implications for diagnosis and management of myofascial pain complaints in the craniofacial and neck region. PMID- 15895834 TI - Pain patterns and mandibular dysfunction following experimental trapezius muscle pain. AB - AIMS: To clarify the effects of experimental trapezius muscle pain on the spread of pain and on jaw motor function. METHODS: In 12 male subjects aged 25 to 35 years, experimental pain was induced in the superior border of the trapezius muscle by injecting 0.5 mL of hypertonic (6%) saline. The control infusion consisted of a 0.5-mL isotonic (0.9%) saline solution. Pain intensity was evaluated on a visual analog scale (VAS). An experimental (EX) and a control (CT) injection were administered to the subjects in a randomized sequence. RESULTS: Pain intensity as scored on the VAS increased immediately after the EX injection and decreased gradually after reaching a peak of 68.0 +/- 16.1 mm at 60 seconds after injection. The VAS scores in the EX condition were significantly higher than after the CT condition from 30 to 330 seconds after injection (P < .05, analysis of variance [ANOVA]). Mean (+/- SD) maximal unassisted mouth opening before injection in the EX condition was 54 +/- 5.7 mm and decreased immediately after the injection, reaching a low of 47.8 +/- 5.1 mm. A gradual recovery to normal was then observed. This reduction of mouth opening in the EX condition was significant compared with the CT condition from immediately after the injection to 60 seconds after the injection (P < .05, ANOVA). According to the subjects, pain spread most often to the infra-auricular zone (n = 6), and the posterolateral part of the neck (n = 10). CONCLUSION: The present results suggest that experimental trapezius muscle pain can spread over a wide area and is also accompanied by a temporary reduction of mouth opening. PMID- 15895835 TI - Generalized pain and pain sensitivity in community subjects with facial pain: a case-control study. AB - AIMS: To investigate the existence of pain outside the facial area as well as pain sensitivity in a population-based sample of 34-year-old subjects with facial pain. METHODS: Fifty-two facial pain cases (10 men, 42 women) and 52 pain-free controls (10 men, 42 women) included in the Northern Finland Birth Cohort of 1966 underwent a clinical musculoskeletal examination. Pain outside the facial area during the week prior to the examination was defined by means of a pain drawing. Eighteen fibromyalgia points were palpated in response to digital palpation with an algometer. Pressure pain thresholds were measured from the dorsal side of the wrist and from the highest points of the temporalis muscles. RESULTS: Compared to controls, pain cases reported significantly more pain in areas outside the face, with the exception of the shoulder and lower back. The number of painful fibromyalgia points was significantly higher in cases than in controls. Mean pressure pain thresholds were slightly lower in cases than in controls; the difference was significant in the left wrist. CONCLUSION: Subjects with facial pain reported more pain and had more mascular tenderness outside the facial area compared to controls. Pain symptoms outside the facial area should be assessed in patients seeking treatment for facial pain, and they should be taken into account when treatment is planned. PMID- 15895836 TI - Cyclic effects on experimental pain response in women with temporomandibular disorders. AB - AIMS: Since cyclic effects on experimental pain response in women with temporomandibular disorders (TMD) have not been adequately studied, the aim of this study was to assess variations in experimental pain response at 4 phases of the menstrual cycle. METHODS: Eighteen normally cycling women with TMD, 25 women with TMD and taking oral contraceptives (OC), 25 normally cycling pain-free controls, and 26 pain-free controls taking OC underwent 3 experimental pain procedures at 4 phases during each of 3 menstrual cycles. These procedures included algometer palpations at fixed amounts of pressure and pressure pain thresholds at several body sites, and an ischemic arm pain task. Repeated measures analysis of variance was used to compare cycle phase, TMD group, and OC status differences in experimental pain response. RESULTS: Significant phase related differences were seen for palpation intensity measures (P values < .05). Normally cycling women with TMD showed higher palpation pain intensity at menses and midluteal phases, while women with TMD taking OC showed stable palpation pain intensity ratings at menses, ovulatory, and midluteal phases, with increased intensity at the late luteal phase. TMD subjects had greater palpation pain and ischemic pain intensity and lower pressure pain thresholds compared to controls. CONCLUSION: Phase-related differences in experimental pain response were not strong and were more often found for experimental stimuli with greater clinical relevance (ie, palpation pain) compared with an ischemic pain task. PMID- 15895837 TI - Prevalence of temporomandibular pain and subsequent dental treatment in Swedish adolescents. AB - AIMS: To assess the prevalence of temporomandibular disorder (TMD) pain in Swedish adolescents, to evaluate whether there are differences in TMD pain in regard to age, gender, and place of residence (urban or rural), and to evaluate treatment for TMD pain and compare it with documented treatment for teeth with caries. METHODS: All youths aged 12 to 19 years in Ostergotland County were eligible to participate in the investigation, which took place at public dental clinics during annual examinations in 2000. The subjects were asked 2 questions: (1) "Do you have pain in your temples, face, jaw joint, or jaws once a week or more?" and (2) "Do you have pain when you open your mouth wide or chew once a week or more?" Two hundred dental records of patients with TMD pain and 100 records of patients with documented caries were randomly selected from the population. These records were used to compare the treatment of TMD pain to the treatment of caries. RESULTS: Among the 28,899 youths participating, 4.2% reported TMD pain. The prevalence increased with age, and a significant difference was seen between boys (2.7%) and girls (6.0%) (P < .001). In 43% of the patients with TMD pain, the TMD condition was noted in the record. TMD related treatment was given to 34% of patients with TMD pain in dental clinics, whereas 100% of the patients with caries received restorative treatment. The most common treatment modalities for the TMD group were occlusal splints and information about TMD. CONCLUSION: The prevalence of self-reported TMD pain was relatively low, increased with age, and was higher among girls than boys. One third of the patients with TMD pain received some form of TMD treatment in the dental clinics. PMID- 15895838 TI - Controlled assessment of the efficacy of occlusal stabilization splints on sleep bruxism. AB - AIMS: To assess the efficacy of occlusal stabilization splints in the management of sleep bruxism (SB) in a double-blind, parallel, controlled, randomized clinical trial. METHODS: Twenty-one participants were randomly assigned to an occlusal splint group (n = 11; mean age = 34.2 +/- 13.1 years) or a palatal splint (ie, an acrylic palatal coverage) group (n = 10; mean age = 34.9 +/- 11.2 years). Two polysomnographic recordings that included bilateral masseter electromyographic activity were made: one prior to treatment, the other after a treatment period of 4 weeks. The number of bruxism episodes per hour of sleep (Epi/h), the number of bursts per hour (Bur/h), and the bruxism time index (ie, the percentage of total sleep time spent bruxing) were established as outcome variables at a 10% maximum voluntary contraction threshold level. A general linear model was used to test both the effects between splint groups and within the treatment phase as well as their interaction for each outcome variable. RESULTS: Neither occlusal stabilization splints nor palatal splints had an influence on the SB outcome variables or on the sleep variables measured on a group level. In individual cases, variable outcomes were found: Some patients had an increase (33% to 48% of the cases), while others showed no change (33% to 48%) or a decrease (19% to 29%) in SB outcome variables. CONCLUSION: The absence of significant group effects of splints in the management of SB indicates that caution is required when splints are indicated, apart from their role in the protection against dental wear. The application of splints should therefore be considered at the individual patient level. PMID- 15895839 TI - Bite force measurement in awake rats: a behavioral model for persistent orofacial muscle pain and hyperalgesia. AB - AIMS: To test the hypotheses that masseteric inflammation produces a reduction of mean bite force and success rates and that classical anti-inflammatory agents prevent inflammation-induced changes in bite force. METHODS: Rats were initially trained to produce a bite force greater than 400 g. Once the rats attained above 70% of successful responses in a 10-minute test period, the bite force required for reinforcement was increased gradually to the target force of 1.3 kg. Seven trained rats received bilateral masseteric injections of complete Freund's adjuvant (CFA; 50 microL in isotonic saline). The mean number of attempted bites, the percentage of correct responses, and the bite force measured before and 1, 2, 3, 7, 10, and 14 days following the CFA injection were compared. Five additional trained rats were injected with the same volume of vehicle control. Other rats (n = 10) were treated with anti-inflammatory agents before and after the CFA injection. RESULTS: Intramuscular CFA, but not the vehicle, produced a significant reduction of mean bite force and success rate at days 1, 2, and 3. Bite force and success rate gradually increased; they returned to baseline by 14 days. The CFA-mediated reduction of bite force and success rate was prevented in rats treated with anti-inflammatory agents administered intraperitoneally (dexamethasone, n = 5, or indomethacin, n = 5, 4 mg/kg). CONCLUSION: These results provide further evidence that bite force measurements in awake rats can be a useful method for the study of inflammatory muscle hyperalgesia. PMID- 15895840 TI - The diagnosis of burning mouth syndrome represents a challenge for clinicians. AB - AIMS: To evaluate the occurrence of professional delay in the diagnosis, referral, and treatment of patients with burning mouth syndrome (BMS). METHODS: Fifty-nine patients (51 women and 8 men; average age, 60.5 years; age range, 32 to 88 years) diagnosed with BMS at our institution were retrospectively studied. Data were collected about the onset of oral symptoms, consultations with medical and dental practitioners, and misdiagnosis before definitive BMS diagnosis and treatment. RESULTS: The average delay from the onset of the symptoms to definitive diagnosis was 34 months (range, 1 to 348 months; median, 13 months). The average number of medical and dental practitioners consulted by each patient over this period and who initially misdiagnosed BMS was 3.1 (range, 0 to 12; median, 3). Candidiasis and aspecific stomatitis were the most frequent misinterpretations of the symptoms before appropriate referral. In about 30% of cases, no diagnosis of the oral symptoms was made or explanation given. CONCLUSION: Professional delay in diagnosing, referring, and appropriately. managing BMS patients occurred frequently in the group studied. No significant differences were found in the number of medical and dental practitioners who were consulted. Emphasis must therefore be placed upon educational efforts to improve health care providers' awareness of BMS. This should increase the rate of recognition and appropriate referral or treatment of patients with chronic orofacial pain due to BMS. PMID- 15895841 TI - Metabolic syndrome and risk of isolated ST-T abnormalities and type 2 diabetes in Japanese male office workers. AB - Using a modified National Cholesterol Education Program (NCEP) definition of the metabolic syndrome (MS) with body mass index instead of waist circumference, we examined the associations of the MS with the risk of developing ST-T abnormalities in 3405 Japanese men aged 35-59 yr who did not have a history of cardiovascular disease or ST-T abnormalities. Of 3405 participants, 3166 men without type 2 diabetes (as diagnosed with the revised criteria of American Diabetes Association) also constituted a non-diabetic cohort. Examinations including electrocardiogram and fasting plasma glucose were repeated annually for 7 subsequent years. The subjects were classified as having ST-T abnormalities or type 2 diabetes when evidence of either of these disorders was found during at least 2 consecutive annual examinations. After adjustment for potential risk factors, the relative risks of ST-T abnormalities were 1.0 (referent), 2.66, 3.07, 4.27, and 8.40 for the presence of 0, 1, 2, 3, and > or =24 components of the MS, respectively (P for trend <0.001). The corresponding results for the risk of type 2 diabetes were 1.0 (referent), 3.49, 7.45, 15.00, and 24.04 (P for trend <0.001). The estimated incident rates for men in the low-WBC count (<7.3 x 10(9) cells/L)/no MS, high-WBC count (> or =7.3 x 10(9) cells/L)/no MS, low-WBC count/yes MS, and high-WBC count/yes MS were 3.4%, 4.6%, 7.4%, and 13.1% for ST-T abnormalities, respectively and were 3.6%, 7.1%, 18.0%, and 27.2% for type 2 diabetes, respectively. The respective multivariate-adjusted relative risks were 1.0 (referent), 1.26, 2.07, and 3.45 for ST-T abnormalities and were 1.0 (referent), 1.75, 5.14, and 6.90 for type 2 diabetes. A modified NCEP MS definition predicts ST-T abnormalities and type 2 diabetes. WBC count adds clinically important information to new-onset ST-T abnormalities and type 2 diabetes. PMID- 15895842 TI - Assessment of respirable dust and its free silica contents in different Indian coalmines. AB - Assessment of respirable dust, personal exposures of miners and free silica contents in dust were undertaken to find out the associated risk of coal workers' pneumoconiosis in 9 coal mines of Eastern India during 1988-91. Mine Research Establishment (MRE), 113A Gravimetric Dust Sampler (GDS) and personal samplers (AFC 123), Cassella, London, approved by Director General of Mines Safety (DGMS) were used respectively for monitoring of mine air dust and personal exposures of miners. Fourier Transform Infra-red (FTIR) Spectroscopy determined free silica in respirable dusts. Thermal Conditions like Wet Bulb Globe Temperature (WBGT) index, humidity and wind velocity were also recorded during monitoring. The dust levels in the face return air of both, Board & Pillar (B&P) and Long Wall (LW) mining were found above the permissible level recommended by DGMS, Govt. of India. The drilling, blasting and loading are the major dusty operations in B&P method. Exposures of driller and loader were varied between, 0.81-9.48 mg/m3 and 0.05-9.84 mg/m3 respectively in B&P mining, whereas exposures of DOSCO loader, Shearer operator and Power Support Face Worker were varied between 2.65-9.11 mg/m3, 0.22-10.00 mg/m3 and 0.12-9.32 mg/m3 respectively in LW mining. In open cast mining, compressor and driller operators are the major exposed groups. The percentage silica in respirable dusts found below 5% in all most all the workers except among query loaders and drillers of open cast mines. PMID- 15895843 TI - Effects of pesticides on the peripheral and central nervous system in tobacco farmers in Malaysia: studies on peripheral nerve conduction, brain-evoked potentials and computerized posturography. AB - We examined the effects of pesticides on the central and peripheral nervous system in the setting of a tobacco farm at a developing country. Maximal motor and sensory nerve conduction velocities (MCV and SCV, respectively) in the median, sural and tibial nerves, postural sway, and brain-evoked potentials (auditory event-related and visual-evoked potentials) were measured in 80 male tobacco farmers and age- and sex-matched 40 controls in Kelantan, Malaysia. Median SCV (finger-wrist) in farmers using Delsen (mancozeb, dithiocarbamate fungicide), who showed significant decrease of serum cholinesterase activities, were significantly lower compared with the controls. Sural SCV in farmers using Fastac (alpha-cypermethrin, pyrethroid insecticide) and median MCV (elbow-wrist) in farmers using Tamex (butralin, dinitroaniline herbicide) were significantly slowed compared with their respective controls. In Delsen (mancozeb, dithiocarbamate) users, the power of postural sway of 0-1 Hz was significantly larger than that in the controls both in the anterior-posterior direction with eyes open and in the right-left direction with eyes closed. The former type of sway was also significantly increased in Tamaron (methamidophos, organophosphorus insecticide) users. In conclusion, nerve conduction velocities and postural sway seem to be sensitive indicators of the effects of pesticides on the central and peripheral nervous system. PMID- 15895844 TI - Cigarette smoking and the risk of the metabolic syndrome in middle-aged Japanese male office workers. AB - To examine the association between cigarette smoking and the risk of the metabolic syndrome (MS), 3649 Japanese male office workers aged 35-59 yr who did not have a history of cardiovascular disease were enrolled in this study. 2994 men without the MS at entry were followed up over a 7-yr period. A modified National Cholesterol Education Program definition with body mass index instead of waist circumference was used for the MS. With adjustment for age, family history of diabetes, alcohol intake, and regular physical activity, the odds ratios of the MS were 1.0 (referent), 1.30 (95% confidence interval (CI), 1.00-1.68), 1.07 (95% CI, 0.82-1.39), 1.17 (95% CI, 0.88-1.56), and 1.66 (95% CI, 1.24-2.20) for never smokers, ex-smokers, and those who smoked 1-20 cigarettes/d, 21-30 cigarettes/d, and > or =31 cigarettes/d, respectively (P for trend for current smokers only =0.006). As for the risk of developing the MS, the respective multivariate-adjusted hazard ratios of developing the MS were 1.0 (referent), 1.43 (95% CI, 1.14-1.79), 1.14 (95% CI, 0.91-1.44), 1.45 (95% CI, 1.14-1.84), and 1.59 (95% CI, 1.24-2.05) (P for trend for current smokers only =0.001). Among men without the MS at entry, body weight gain over 7 yr, compared with never smokers, was significantly higher in smokers who quit smoking. It is important for the prevention of the MS not only to quit smoking but also to prevent weight gain after smoking cessation. PMID- 15895845 TI - Chronic laryngitis in glassblowers. AB - Glassblowers are exposed to numerous physical and chemical noxious factors at their workplace. Mucosal layer of upper aerodigestive tract (nose, oral cavity, pharynx and larynx) is particularly vulnerable to the influence of these factors. Inhalation of hot gases, dust particles, and oral contact with glassblower's pipe are the most important factors that can cause chronic inflammation of the upper respiratory tract. The aim of this study was to examine the prevalence of chronic laryngitis in the group of glassblowers and control group of workers and to examine whether the observed differences depend on the membership in these two groups, on the duration of exposure, or on both factors. The results of the study show significantly higher prevalence of chronic laryngitis among glassblowers than among control group of workers employed in the same plant. On the other hand, the duration of exposure was not statistically significant factor for chronic laryngitis. The exposure to risk factors on the working place represents an independent risk factor for the occurrence of chronic laryngitis among glassblowers, when adjusted for other possible confounders, such as age, years of service, smoking habits and alcohol consumption. PMID- 15895846 TI - Two-week oral toxicity study of 1,4-Dichloro-2-nitrobenzene in rats and mice. AB - Subacute toxicity of 1,4-dichloro-2-nitrobenzene (DCNB) was examined by feeding F344 rats and BDF1 mice of both sexes a diet containing DCNB at 625, 1250, 2500, 5000 or 10,000 ppm (w/w) for 2 weeks. All DCNB-fed rats survived to the end of the 2-week administration period, but 2 male and 6 female mice fed 10,000 ppm died during this period. The subacute toxicity was characterized by lesions affecting the liver, kidney, testis and hematopoietic system. The liver was the most responsive to DCNB, as evidenced by a dose-related increase in relative liver weight in rats and mice and centrilobular hypertrophy of hepatocytes in mice. An alteration in liver-associated lipid metabolism was suggested from the concomitant increases in serum concentrations of total cholesterol and phospholipid. A lower confidence limit of the benchmark dose yielding the response with 10% extra risk (BMDL10) for the relative liver weight indicated that rats were more responsive to DCNB than mice. The kidney lesion was characterized by alpha2upsilon-globulin-accumulated hyaline droplets in the renal tubular epithelial cells only in male rats, as indicated by positive anti alpha2upsilon-globulin immunohistochemical staining. Testicular and hematopoietic lesions appeared at higher dose levels than did the liver and kidney lesions. PMID- 15895847 TI - Neurobehavioral changes of shipyard painters exposed to mixed organic solvents. AB - This study was conducted to evaluate neurobehavioral changes arising from occupational exposure to organic solvents among shipyard painters and to establish whether a dose-effect relation existed where there was any observed impairment of neurobehavioral performance by running the test of Simple Reaction Time (SRT), Symbol Digit Substitution (SD), and Finger Tapping Speed (FT) with a computer-assisted neurobehavioral test battery. The study group consisted of 180 shipyard painters and 60 reference workers. The workers answered a self administered questionnaire on occupational, medical history, and demographic characteristics including age, work duration, education level, and quantity and frequency of alcohol and smoking, and performed three psychometric tests on the Korean Computerized Neurobehavioral tests. To estimate cumulative exposure level, samples of ambient air on 61 painters were analyzed using a gas chromatograph. Shipyard painters and the reference group showed significant differences in the results of test of SD, FT of dominant hand, and FT of non-dominant hand. The test results of SD of shipyard painters also showed significant difference by duration of work. This suggests that occupational exposure of organic solvent could induce neurobehavioral changes in the shipyard painters. Therefore an objective neurobehavioral tests recommended on evaluating neurobehavioral performance of long-term solvent-exposed shipyard workers. PMID- 15895848 TI - FirstAid knowledge among industry workers in Greece. AB - Occupational accidents cause serious consequences. Providing first aid (FA) in the workplace can reverse unpleasant results. Aim of our study was to evaluate the knowledge on FA of industry workers through a questionnaire. Our sample constituted of 236 workers, 40 (Group A) were trained on FA by a team of doctors authorised by the Hellenic National Centre of Emergency Help, while 196 had never have a formal training on FA (Group B). In Group A the mean value of correct answers (performance score) was higher than Group B (62.7 and 50.4 respectively), while the total sample of workers ranged from 21.1 to 84.2. Group A replied correctly about normal values (NVs) of pulse, arterial blood pressure (BP) and breaths at 67.5%, 35% and 47.5% respectively. Group B replied at 32.8%, 18.4% and 30.6% on the corresponding NVs. In Group A, 95% answered properly about the definition of Cardiopulmonary Rehabilitation (CPR) but only 25% were acquaint with the necessary number of chest compressions and 32.5% with the number of necessary mouth-to-mouth ventilations to practice in case of CPR. In Group B, the corresponding percentages were 72.4%, 26% and 36.7%. Only 5% from Group A and 1% from Group B answer correctly and simultaneously about estimation of vital signs and CPR practice. In conclusion, the vast majority of workers are unable to provide efficient FA as long as they can't estimate vital signs and provide satisfactory CPR. PMID- 15895849 TI - Simultaneous determination of 8-hydroxydeoyguanosine, a marker of oxidative stress, and creatinine, a standardization compound, in urine. AB - Recently, H. Kasai reported an automatic, precise method of 8 hydroxydeoxyguanosine (8-OH-dG) analysis in urine by high performance liquid chromatography coupled to an electrochemical detector (HPLC-ECD). It is based on a cleaning-up step by anion-exchange chromatography and a further purification step using reverse phase chromatography before detection, by the ECD. In this communication, we report a method for the simultaneous determination of 8-OH-dG and creatinine, an internal standard for normalizing the excretion of 8-OH-dG in urine. PMID- 15895850 TI - Psychophysically determined asymmetrical lifting capabilities for different frequencies and containers. AB - Ten young male participants were tested their psychophysically determined 4-h maximum acceptable weight of lifting (MAWL) of 90-degree asymmetric lifting for three lifting frequencies (1 lift/min, 2 lifts/min, 4 lifts/min) and three lifting containers (50 x 35 x 15 cm, 70 x 35 x 15 cm, 50 x 50 x 15 cm). The results showed that the MAWL decreased with increasing frequency and container width or length dimension, while the interaction effect of frequency and container on MAWL was not significant. The MAWL ranged from 13.7 kg to 18 kg for the nine (3 frequencies x 3 containers) lifting conditions. When averaged across the levels of the other independent variable, the MAWL decreased by 7.4% and 16.1% for 2 lifts/min and 4 lifts/min as compared with the MAWL of 1 lift/min, respectively, and decreased by 1.6% and 9.4% for 70 x 35 x 15 cm and 50 x 50 x 15 cm as compared with the MAWL of 50 x 35 x 15 cm, respectively. Additionally, the discrepancies between the MAWL data and the recommended weight limits derived from the revised NIOSH equation were discussed. PMID- 15895851 TI - A case of sick building syndrome in a Japanese office worker. AB - The adverse health effects caused by indoor air pollution are termed "sick building syndrome". We report such a patient whose symptoms appeared in the workplace. A 36-year-old female office worker developed nausea and headache during working hours in a refurbished office. After eight months of seeking help at other clinics or hospitals without improvement, she was referred to our hospital. At that time she reacted to the smells of various chemicals outside of the office building. Biochemical findings were all within normal ranges. Specific IgE antibody to cedar pollen was positive and the ratio of TH1/TH2 was 4.5. In the Eye Tracking Test (ETT), vertical eye movement was saccadic. Her anxiety level was very high according to the State-Trait Anxiety Inventory (STAI) questionnaire. Subjective symptoms, ETT findings and anxiety levels on STAI gradually improved during two years of follow-up. One year after the onset of her illness, the formaldehyde concentrations in the building air ranged from 0.017 0.053 ppm. Even though relatively low, chemical exposure from building materials such as formaldehyde induced a range of symptoms. Also, lack of recognition by superiors and doctors that sick building syndrome might have been the source of her illness coupled with her high state of anxiety may have exacerbated her symptoms and led to the onset of multiple chemical sensitivity. Thus psychosocial factors may contribute to sick building syndrome in the workplace. PMID- 15895852 TI - An autopsy case of primary pericardial mesothelioma in arc cutter exposed to asbestos through talc pencils. AB - An autopsy case of a primary pericardial mesothelioma in a 53-year-old arc cutter is reported. He had often had the chance to inhale dust generated by sharpening the slate pencils composed of talc. He was admitted for heart failure due to pericardial tumor, but later died. The tumor was mainly located on the pericardium with a thickness of about 2.5 cm. Small nodular disseminations were observed in the left parietal pleura. Microscopically, tumor cells were epithelial-like and rich in histochemical demonstrable hyaluronic acid. Findings of immunohistochemical markers revealed keratin (+), EMA (+), calretinin (+), and CEA (-), which were characteristics of mesothelioma of epithelial type. The number of asbestos bodies (AB) in the lung parenchyma was increased (2026 AB/gram dry lung tissue). Subsequent transmission electron microscopic examination equipped with an energy dispersive X-ray analyzer revealed that the fibers identified in the lungs were fibrous talc and actinolite. These findings suggested that this patient had been occupationally exposed to asbestos contaminated in the talc pencils, which induced the development of primary pericardial mesothelioma. PMID- 15895853 TI - Some engineering countermeasures to reduce exposure to welding fumes and gases avoiding occurrence of blow holes in welded material. AB - Recently, open-type push-pull ventilation systems have been widely employed as effective substitutes for the conventional local exhaust ventilation system, and have prevailed at many welding workshops in Japan. In this study, the effect of the uniform velocity on carbon dioxide (CO2) shielding arc welding was examined by laboratory experiments. The ventilation system examined in the experiments successfully fulfilled the requirement for open-type push-pull ventilators prescribed in Japanese regulations (ordinances). It was proved that the velocity at any points in the capture zone fell in the range of 50 to 150% of the average capture zone velocity. Welding defects could be avoided by controlling the flow rate of shielding gas. Unless the capture velocity exceeded a 0.8 m/s, the formation of blow-holes in the welded metal could be prevented at the shielding gas flow rate of 20 L/min. If the flow rate was provided at 30 L/min and 40 L/min, blow-holes didn't form at the capture velocity of 1.2 m/s and 1.6 m/s, respectively. At a capture velocity of faster than 0.3 m/s, the fume concentration at welder's breathing zone was reduced to a level below the limit values: ACGIH (TLV) and Japan Welding Engineering Society (CLV#). These data are important for designing open-type push-pull ventilation in the welding workshop. The other engineering countermeasures currently employed in the welding work in Japan, such as fume collecting torch and general ventilation, are also concerned in this report. #: Control Limit Value. PMID- 15895854 TI - [Neurobiological basis of human recognition of facial emotion]. AB - In the review of modern data and ideas concerning the neurophysiological mechanisms and morphological foundations of the most essential communicative function of humans and monkeys, that of recognition of faces and their emotional expressions, the attention is focussed on its dynamic realization and structural provision. On the basis of literature data about hemodynamic and metabolic mapping of the brain the author analyses the role of different zones of the ventral and dorsal visual cortical pathway, the frontal neocortex and amigdala in the facial features processing, as well as the specificity of this processing at each level. Special attention is given to the module principle of the facial processing in the temporal cortex. The dynamic characteristics of facial recognition are discussed according to the electrical evoked response data in healthy and disease humans and monkeys. Modern evidences on the role of different brain structures in the generation of successive evoked response waves in connection with successive stages of facial processing are analyzed. The similarity and differences between mechanisms of recognition of faces and their emotional expression are also considered. PMID- 15895855 TI - [Sensory integration: hierarchy and synchronization]. AB - This is the first in the series of mini-reviews devoted to the basic problems and most important effects of attention in terms of neuronal modeling. We believe that the absence of the unified view on wealth of new date on attention is the main obstacle for further understanding of higher nervous activity. The present work deals with the main ground problem of reconciling two competing architectures designed to integrate the sensory information in the brain. The other mini-reviews will be concerned with the remaining five or six problems of attention, all of them to be ultimately resolved uniformly in the framework of small modification of dominant model of attention and memory. PMID- 15895856 TI - [Cognitive set depends on the involvement of the ventral and dorsal visual systems]. AB - Healthy adults were examined in three series of experiments with formation of an unconscious visual set: 1) the set was formed by repeated presentation of pairs of unequal circles (control); 2) an additional task of recognition of words/pseudowords was introduced into the context of the set-forming trials; 3) in the task additionally introduced, a subject had to spatially localize a certain target letter in a letter matrix. Scores of stability of the visual set to circles were compared. Coherence of the cortical electric activity in the alpha band was analyzed. We revealed a significant decrease in stability (rigidity) of the nonverbal visual set in the series with the additional task of spatial localization of the target stimulus. On the contrary, the set rigidity increased in the series with additional recognition of the verbal stimulus. EEG coherence patterns and behavioral data suggest that successful cognitive performance that demands dynamic situation-dependent shifts of unconscious sets takes place under conditions of alternation of tasks involving, predominantly, either the ventral ("what?") or dorsal ("where?") visual streams and, respectively, anterior or posterior systems of selective attention. PMID- 15895857 TI - [EEG pattern as an instrument for evaluation of neurophysiological mechanisms underlying adaptation disorders]. AB - The method of statistical evaluation of probable interaction between the main constituents of bioelectrical activity of the brain (BEA) enables to reveal the character of intercentral relations both under normal conditions and at adaptation disorders of different severance. Several types of statistic interactions have been revealed between EEG components (SSIC) correlating with normal interrelations between cortex and subcortex, level of preferential involvement of non-specific brain formations (cortex, thalamic, hypothalamic and brain-stem) into the process, and the degree of instability of neuro-dynamic processes. The findings obtained have shown the degree of expression of adaptation disorders to be connected with gradual destruction of "functional nuclear" within alpha-range, amplification of statistical interrelations with beta-EEG and teta-EEG components and formation of pathological nuclear structures within the ranges aforesaid. PMID- 15895858 TI - [EEG correlates of individual differences in performance efficiency of students during examination stress]. AB - Performance of cognitive tests and EEG spectral power were evaluated in 39 students aged from 19-21 years in two conditions: during common educational process and immediately before examination (stress condition). Before examination, the performance was better in subjects with low level of spectral density in the delta band (in the occipital, parietal, central and frontal cortical areas) and high level of the alpha-rhythm spectral density in all the cortical areas, A decrease in performance scores before examination was correlated with an increase in the delta activity (in the right frontal and temporal cortical areas) and rise of the anxiety level (tested by Spielberger). PMID- 15895859 TI - [Brain mechanisms of the semantic analysis of words-homonyms]. AB - The brain mechanisms of semantic comprehension of a word were comparatively studied in three experimental conditions: simple perception of the nuclear value of a word-homonym determined by the preceding context, perception of its circumferential value, and during active semantic analysis of these values. It was shown that the amplitude of the evoked potential component P200 was correlated with complexity of the semantic analysis. A decrease in this amplitude under conditions of complication of semantic problem was associated with an increase in the activities of the caudate nucleus and hippocampus and a parallel slight decrease in the activity of cortical areas. PMID- 15895860 TI - [Transcranial magnetic stimulation in research of emotion in the healthy and patients with epilepsy]. AB - The main aim of this work was by using transcranial magnetic stimulation to investigate mechanisms of interhemispheric organization the emotion in the healthy and patients with epilepsy. The research was carried out on three groups: the first and second groups of healthy and third group of the patients with idiopatical epilepsy. The first and third groups received transcranial magnetic stimulation on right and left frontalis area. The second group was control (sham transcranial magnetic stimulation). Is shown, that transcranial magnetic stimulation of right frontalis area increases the examining time on negative photos and decreases on positive photos. Transcranial magnetic stimulation of left frontalis area in the healthy and patients with epilepsy increases the examining time on positive photos and decreases on negative photos. The right hemisphere at the healthy and patients with epilepsy in the greater degree is connected to negative marks of emotions, and left hemisphere with positive marks of emotions. PMID- 15895861 TI - [Effects of fluoxetine and its complexes with glycyrrizhinic acid on behavior and brain monoamine levels in rats]. AB - Effects of serotonin uptake inhibitor fluoxetine (F) and it's complexes with glycyrrizhinic acid (GA) in molar proportions 1GA : 1F (FGA-1) and 4GA : 1F (FGA 4) on rat behavior in elevated plus-maze and brain monoamine concentrations were studied. Drugs (25 mg/kg) were administered per os 1 h before investigations. F treated rats showed increased anxiety and reduced locomotor activity, whereas FGA 1 and FGA-4 had no effects on the behaviors. None of the compounds modified brain tissue serotonin content, but all of them decreased the level of its metabolite 5 hydroxyindole-3-acetic acid level in the hypothalamus, and FGA-4 also decreased it in the cortex. Noradrenaline levels were increased in the hypothalamus of rats treated with F in both combinations with GA. In the striatum, F increased dopamine and its metabolite DOPAC levels, but their ratio (an indicator of the neurotransmitter turnover) was not altered by this drug. Unlike F, FGA-1 significantly activated dopamine turnover in the striatum. The data obtained suggested that application of F in complexes with GA significantly modified the drug behavioral effects and these alterations may be related to specific effects of the pure compound and its complexes on the functions of the brain monoaminergic systems that regulate investigated behavior. PMID- 15895862 TI - [Long-lasting behavioral effects of chronic neonatal treatment with ACTH (4-10) analogue semax in white rat pups]. AB - It is well known that ACTH/MSH-like peptides (melanocortins) have neurotrophic and neuroprotective effects on the central and peripheral nervous systems in the early postnatal life. The aim of present work was to study consequences of the ACTH (4-10) analogue Semax influence on the developing brain. The work was carried out in white rat pups. The peptide (0.05 mg/kg, i/p) was injected daily on the 8th-21st postnatal days. Delayed long-lasting effects of such treatment on animal behavior were revealed. At the age of four to eight weeks, Semax-treated rats displayed elevated exploratory activity, decreased anxiety level and improved passive avoidance conditioning. The results suggest that neonatal Semax administration modulates the development of the central nervous system. PMID- 15895863 TI - [Training and extinction of passive avoidance in mice lacking monoamine oxidase A]. AB - The passive avoidance learning and memory trace retention in mice lacking monoamine oxidase A (MAO A) and control C3H strain were analyzed. It is shown that mice of both strains were well the passive avoidance learners. A delay of the memory trace extinction was found in lacking MAO A mice as compared with control C3H strain. Mice with a genetic MAO A knockout showed decreased amounts of transitions, rearings, looking to a dark compartment and appearing from it. These findings seem to reflect more expressed fear reaction to a dangerous compartment and increased anxiety promoting longer retention of memory trace on a high level of retrieval. PMID- 15895864 TI - [Poststressor change in attractiveness of mice odourgens]. AB - It was shown, that under physiological conditions, intact noninbred mice or high inbred mice-testers in reaction of preference-avoidance prefer volatile components secreted by mice of a genetically identical group. After exposure to stress factors, mice release with urine the volatile components, which are more preferable by testers than the volatile secretion of intact mice indepedently of the genotype of mice. PMID- 15895865 TI - [Features of the c-Fos gene expression along the hippocampal rostro-caudal axis in common voles after rapid spatial learning]. AB - The levels of the Fos protein expression in neurons was used as an index of transcription activation in the hippocampus of common voles (Microtus arvalis Pall.) after their rapid spatial learning. Fos-positive cells were stained and calculated in 20 brain sections along hippocampal rostro-caudal axis. Voles (learning group) were trained in a modified 8-arm radial maze to find the entry to the home cage through a target arm (6 trials per session, 2-hour session). The animals were pretrained to enter the home cage through an arm isolated from the maze. Animals of active control group continued entering the home cage through the isolated arm, and animals of the passive control group were taken for the Fos immunohistochemistry from the home cage. Both in the learning group and active control group, a significant increase in c-Fos expression was shown in all the examined areas (CA1, CA3 and the dentate gyrus) as compared to the passive control. A significant increase in the number of c-Fos positive neurons was observed in the caudal hippocampus of the learning animals as compared to the active control, however, no differences were found in the rostral part. The maximum effects were observed in the dentate gyrus and the CA3 field. The results suggest a functional rostro-caudal inhomogeneity of the vole's hippocampus in the spatial learning task. PMID- 15895866 TI - [The role of corticotropin-releasing hormone in alteration of adaptive behavior of the active and passive rats after inescapable stress]. AB - Effects of corticotropin-releasing hormone (CRH) on the formation of post-stress psychopathology were studied using of two genetic strains KHA (Koltushi high Avoidance) and KLA (Koltushi low Avoidance) selected on high or low acquisition of active avoidance, respectively. These strains are characterized by higher (KHA) and lower (KLA) behavioral activity in open field and adopted, respectively, active and passive strategies in stressful conditions. A widely used experimental paradigm of learned helplessness where behavioral depression was produced by inescapable uncontrollable footshock has been applied in our study. KHA rats demonstrated psychopathology already 1st day following exsposure to the stress faktor, and the depression progressed by the 5th and 10th post stress days. Intranasal application of CRH facilitated the development of depression in active rats. In KLA rats, which originally displayed low exploratory activity associated with high anxiety, the inescapable stress at first enhanced the exploratory behavior but 10 days later these rats displayed a progressive decline of exploration and locomotion. Initially, the application of CRH also enhanced the exploratory behavior in these rats, but to 10th post-stress day promoted development of depressive state. The results suggest that CRH in different ways affects the formation of depressive state in rats with different strategies of adaptive behavior. PMID- 15895867 TI - [Effects of neurotensin on active and passive avoidance performance in rats with lesions of serotoninergic neurons]. AB - After a lesion of serotoninergic neurons performed by administration of 5.7 dihydroxytriptamine into the dorsal raphe nucleus, effects of neurotensin microinjections into the substantia nigra on rat behavior were investigated. Serotoninergic lesions resulted in enhanced fear of rats manifested as an increase in the number of intersignal avoidance reactions and intensification of escape reactions. Neurotensin microinjections into the substantia nigra diminished the neurotoxin action thus increasing the adaptive character of defensive behavior of rats with deficit of functions of serotonin neurons. PMID- 15895868 TI - [Are WAG/Rij rats with genetic absence epilepsy anxious?]. AB - Behavior of susceptible and non-susceptible to audiogenic (convulsive) seizures rats from inbred WAG/Rij strain, genetically predisposed to absence epilepsy, and outbred Wistar strain, genetically not predisposed to absence epilepsy, was compared to assess the level of anxiety (in open field, light-dark choice and elevated plus-maze tests) and the level of depressiveness (in the sucrose consumption and forced swimming tests). Increased level of anxiety was found only in susceptible to audiogenic seizures rats both from WAG/Rij and Wistar strain, but increased level of depressiveness was found only in WAG/Rij strain rats as compared with Wistar rats independently of their susceptibility to audiogenic seizures. Results suggest that increased depressiveness in WAG/Rij strain rats is associated with absence epilepsy but increased anxiety with susceptibility to audiogenic seizures. PMID- 15895869 TI - [Role of filtration in localization of the evoked potential dipoles in the human brain: experiment and simulation]. AB - 3D dipole tracing with 1 ms step of visual evoked potentials recorded from 40 electrodes was performed under exposition of crosses in 5 healthy human subjects. The data on dipole displacement were compared with prediction of the simulation study on distortion of dipole localization by the signal filtration in the low frequency band. These predictions were experimentally confirmed: the effect depends on the degree of filtration (0.1 or 0.5 Hz) and on the latency of EP waves. Localization and strength of P1 dipoles were not changed under filtration, while for later components--N2 and especially P3--they changed significantly. For the improvement of these distortions time constant of the amplification tract must be some times longer than the time of the dipole activity. PMID- 15895870 TI - [Acceptor of action results as a structural functional basis of dynamic stereotype activities of the brain]. AB - The system mechanisms of brain dynamic stereotypes formation are considered. The brain dynamic stereotypes are shown to be formed on the structures of acceptor of action results by dominating motivations and reinforcements. Acceptors of action results are widely spread in brain structures. They are presented in functional systems which form behavioral acts of animals with spreading neural excitations in collaterals of axons of pyramidal tract. Reinforcing excitations form specific architectonic of acceptors of action results, which include brain structures corresponding to modalities of parameters of reinforcements. Dominating motivations, which predict future events, excite molecular engrams of action results which were formed by previous reinforcements. PMID- 15895871 TI - Common variable immunodeficiency: experience in Puerto Rico. AB - Common variable immunodeficiency (CVI) is a primary immunodeficiency characterized by hypogammaglobulinemia and an increased susceptibility to infections. The degree and the type of deficiency of serum immunoglobulins, as well as, the clinical course vary from patient to patient, hence the term "variable". The aim of this report is to describe the clinical characteristics and the response to gammaglobulin therapy of a group of patients with CVI followed at the University Hospital of the Puerto Rico Medical Center. To our knowledge, no data on primary immunodeficiencies in Puerto Rico has been reported in the literature. The study group exhibits specific characteristics as compared to other reported series. PMID- 15895872 TI - Infliximab in Hispanics: characterization of response to infliximab in an ethnic minority with Crohn's disease. AB - PURPOSE: The incidence and prevalence of Crohn's disease (CD) varies geographically and with racial/ ethnic background. The highest frequency of occurrence is in North America and Northern Europe. Incidence is highest among Caucasians, lower in blacks and Hispanics, and lowest in Asians. However in the mid-1980s and 1990s, the incidence and prevalence increased in continental Europe, the Middle East, the Pacific Rim, Africa, and Latin America. An increase in the incidence of CD has been noted in Puerto Rico, although our population differs genetically from other described CD populations. A study in our population showed lower prevalence of ASCA and no NOD2 in our CD patients. Infliximab, a TNFa antibody, is effective in refractory inflammatory CD and in fistulizing disease. Since limited data exists regarding CD in Hispanics, the fastest growing minority group in the United States, we designed this retrospective study with patients treated with infliximab at our institution. We wanted to determine if the response to infliximab in genetically admixed Hispanics differed from that previously reported. METHODS: Baseline characteristics, infusion related information and clinical response was abstracted from medical records. Clinical response was classified as complete response, partial response, and nonresponse. RESULTS: The study included 15 patients treated for refractory inflammatory disease, 9 for fistulizing disease, and 11 for both. The positive response rate was 83%(29/35) and the non response rate was 17%(6/35). Overall the patients with complete, partial, and no response were 13/35(37%), 16/35(46%), and 6/ 35(17%), respectively. No statistically significant association was found between response and disease location. Significant association was found between response and fistula type (p = 0.02). Steroid withdrawal was possible in 21/31 patients (68%). In terms of safety, 9/35 patients (26 %) suffered an adverse reaction, 4 patients required therapy discontinuation. CONCLUSION: This study suggests that infliximab has similar global response, allowance of steroid withdrawal and safety in Hispanics as in other populations. Ethnicity does not seem to influence response rate to infliximab. PMID- 15895873 TI - Recent developments in the therapeutic potential of cannabinoids. AB - OBJECTIVE: To examine the recent evidence that marijuana and other cannabinoids have therapeutic potential. METHODS: Literature published since 1997 was searched using the following terms: cannabinoid, marijuana, THC, analgesia, cachexia, glaucoma, movement, multiple sclerosis, neurological, pain, Parkinson, trial, vomiting. Qualifying clinical studies were randomized, double-blind, and placebo controlled. Selected open-label studies and surveys are also discussed. RESULTS: A total of 15 independent, qualifying clinical trials were identified, of which only three had more than 100 patients each. Two large trials found that cannabinoids were significantly better than placebo in managing spasticity in multiple sclerosis. Patients self-reported greater sense of motor improvement in multiple sclerosis than could be confirmed objectively. In smaller qualifying trials, cannabinoids produced significant objective improvement of tics in Tourette's disease, and neuropathic pain. A new, non-psychotropic cannabinoid also has analgesic activity in neuropathic pain. No significant improvement was found in levodopa-induced dyskinesia in Parkinson's Disease or post-operative pain. No difference from active placebo was found for management of cachexia in a large trial. Some immune system parameters changed in HIV-1 and multiple sclerosis patients treated with cannabinoids, but the clinical significance is unknown. Quality of life assessments were made in only three of 15 qualifying clinical trials. CONCLUSION: Cannabinoids may be useful for conditions that currently lack effective treatment, such as spasticity, tics and neuropathic pain. New delivery systems for cannabinoids and cannabis-based medicinal extracts, as well as new cannabinoid derivatives expand the options for cannabinoid therapy. More well-controlled, large clinical tests are needed, especially with active placebo. PMID- 15895874 TI - RNA interference: a novel and physiologic mechanism of gene silencing with great therapeutic potential. AB - The post-genomics scientific era has evolved rapidly while achieving advanced understanding of the structure and function of the genes responsible for both the phenotypic characteristics of higher organisms and the pathophysiology of several genetic diseases. Researchers in the fields of oncology and infectious diseases have become more convinced of the great potential of molecular biology approaches to further develop highly specific diagnostic and less toxic therapeutic strategies. During the last two decades, approaches for the specific silencing of essential viral genes and cellular oncogenes were evaluated with optimism for developing directed therapies. However, there were drawbacks in the use of antisense oligonucleotides as a practical mechanism of achieving gene silencing both in vitro and in vivo. Recently, a novel role for post-transcriptional gene silencing mediated by double-stranded RNA (dsRNA) was discovered in the experimental model of C. elegans. This mechanism, termed RNA interference (RNAi) has also been found in other eukaryotes, from plants to mammals, including humans. RNAi is presently being explored both in vitro and in vivo in functional genomics studies and possible therapeutic uses due to its highly specific and physiologic mode of gene silencing. This article focuses on the most current information available regarding the RNAi mechanism and its uses in models of cancer and infectious diseases. PMID- 15895875 TI - Effect of three extraction techniques on submitochondrial particle and Microtox bioassays for airborne particulate matter. AB - The effect that three extraction techniques (e.g., Soxhlet, ultrasound and microwave-assisted extraction) have on the toxicity, as measured by submitochondrial particle (SMP) and Microtox assays, of organic extracts was compared from three sources of airborne particulate matter (APM). The extraction technique influenced the toxicity response of APM extracts and it was dependent on the bioassay method, and APM sample source. APM extracts from microwave assisted extraction (MAE) were similar or more toxic than the conventional extraction techniques of Soxhlet and ultrasound, thus, providing an alternate extraction method. The microwave extraction technique has the advantage of using less solvent volume, less extraction time, and the capacity to simultaneously extract twelve samples. The ordering of APM toxicity was generally urban dust > diesel dust > PM10 (particles with diameter < 10 microm), thus, reflecting different chemical composition of the samples. This study is the first to report the suitability of two standard in-vitro bioassays for the future toxicological characterization of APM collected from Puerto Rico, with the SMP generally showing better sensitivity to the well-known Microtox bioassay. PMID- 15895876 TI - The microbial etiologies of diarrhea in hospitalized patients from the Puerto Rico Medical Center Hospitals. AB - The development of diarrhea in hospitalized patients is a frequently encountered clinical problem, which may be due to infectious or non-infectious causes. The purpose of this study was to identify which common community enteric pathogens, if any, are responsible for diarrheal episodes in hospitalized patients. Stool samples from 76 consecutive, hospitalized patients were analyzed utilizing routine bacterial cultures, smears for identification of ova and parasites and Enzyme-Link Immunoadsorbent Assay (ELISA) for enteric bacteria, parasites and viruses. The results obtained demonstrated that the usual community enteric pathogens were not identified as a major cause of nosocomial diarrhea. In hospital-acquired diarrhea, Clostridium difficile toxins assay was the only clinically significant test in the evaluation of these patients. As a result of this study a guideline for the management of this condition in hospitalized patients is presented. PMID- 15895877 TI - Renohepaticopancreatic dysplasia: diagnostic dilemma. AB - We present the case of an eighteen day old baby boy hospitalized with an abdominal mass, renal insufficiency and jaundice. Multiple radiographic, radionuclear and surgical interventions were required to diagnose renohepaticopancreatic dysplasia, also known as Ivemark II syndrome. In spite of aggressive intensive care support, the patient developed multisystemic organ failure and died. Clinical presentation and autopsy findings are presented. PMID- 15895878 TI - [Odontology in Spain: towards the integration to the European high education space]. AB - The European Higher Education Space (EHES) is a project to harmonize all the European University systems in order that all of them will posses a homogeneous structure of undergraduate, graduate and post graduate studies. The time period established to implement this project is year 2010. The present paper describes the establishment of a new career degree model in Spain, the degree in Dentistry that started twenty years ago. Also it has been stated the number of the present Departments and Schools, the access system to Public University Schools of Spain, the evaluation system, and which are the new mechanisms incorporated in the Spanish University system to implement this integration to the EHEE. This is the moment to design the new master guidelines to obtain homogeneous career degrees that may imply more accessibility and mobility of students and professionals. PMID- 15895879 TI - [Towards a hostosian bioethics]. PMID- 15895880 TI - Inherited autoinflammatory syndromes: an expanding new group of chronic inflammatory diseases. PMID- 15895881 TI - Disease-modifying antirheumatic drugs and bone mass in rheumatoid arthritis. AB - This article reviews the effects of DMARDs (including biologic agents) on bone metabolism in rheumatoid arthritis (RA). At present there is no evidence that methotrexate, at least at dosages ranging from 5 to 20 mg/week, negatively affects bone mass as measured by DXA (BMD) as documented in both cross-sectional and longitudinal studies. Most studies of cyclosporine (CyA) use reporting a reduction in erosions and joint damage with no adverse effects on bone, did not measure BMD; CyA treatment is associated with a dose-dependent increase of bone turnover as well as a decrease in both animal and human studies; however, its use in RA setting at a dose < or =5 mg/Kg/ day has so far not been associated with clinical relevant adverse effects on bone metabolism. Anti-TNF-alpha agents, infliximab reduced markers of bone turnover in two longitudinal studies. Data on BMD are not available in RA; nevertheless, an increase in BMD has been documented in spondyloarthropathies with infliximab and etanercept. No clinical data concerning BMD are available on leflunomide as well as on the newer biologic agents (adalimumab, rituximab, anakinra). PMID- 15895882 TI - Repeated infusions of low-dose infliximab plus methotrexate in psoriatic arthritis: immediate benefits are not maintained after discontinuation of infliximab. AB - OBJECTIVE: To assess the long-term efficacy and tolerability of a therapy consisting of infliximab at low dosage plus methotrexate in patients with psoriatic arthritis (PsA). As a second objective, we assessed whether the improvement obtained after 54 weeks of infliximab could be maintained with methotrexate alone. METHODS: A group of 26 patients with peripheral PsA resistant to various DMARDs were treated with infliximab + methotrexate for 54 weeks. RESULTS: The clinical response after the induction period was constant and progressive, with a high percentage of patients achieving an ACR50 response. The ESR and CRP values also declined continuously and gradually, but only CRP returned to normal values. During the follow-up period after 54 weeks, infliximab was stopped and the improvement obtained lasted for 2-6 months. The secondary end point was not achieved, and an extension period was designed. Results at 78 weeks are presented. CONCLUSIONS: Open questions for treating patients with infliximab and methotrexate are the schedule and the length of the administration and how to preserve the improvement obtained after the drug discontinuation. PMID- 15895883 TI - Distinctive effects of G-CSF, GM-CSF and TNFalpha on neutrophil apoptosis in systemic lupus erythematosus. AB - OBJECTIVE: To investigate the influence of culture with G-CSF GM-CSF and TNFalpha on neutrophil apoptosis, comparing neutrophils from SLE patients with rheumatoid arthritis (RA) patients and healthy control subjects. METHODS: Neutrophils were isolated from SLE (n= 10), RA (n= 10) and healthy control subjects (n= 10), and cultured with two different concentrations of G-CSF, GM-CSF and TNFalpha. Proportion of apoptotic neutrophils at T=0, T=2hrs and T=24hrs was measured using FITC-labelled annexinV and flow cytometry. RESULTS: Significantly more neutrophils were apoptotic at T=0 in the SLE subjects than in the other groups (median, range--Control 3.5% (0.3-7.9) SLE 9.5% (2.9-29.1) RA 3.0% (0.4-23.0) p<0.05). Following culture for 24 hours with 1ng/ml G-CSF the proportion of apoptotic neutrophils from SLE subjects was significantly increased (median, range = 51.6% (27.0-84.0) without G-CSF v 66.8% (31.8-89.2) with G-CSF p<0.05). This was not observed with RA or control subjects, in whom the trend was towards inhibition of apoptosis. Similar trends were seen with GM-CSF There was significant induction of apoptosis in SLE neutrophils after 2 hr culture with 1ng/ml TNFalpha (median, range = 2.3% (0.1-8.0) without TNFalpha v 5.2% (1.0 22.4) with TNFalpha). No significant change was seen in the other groups. There was an inverse correlation between total neutrophil count and the degree of induction of apoptosis by G-CSF and GM-CSF, determined at a range of time-points and cytokine concentrations CONCLUSIONS: Neutrophils from SLE patients display resistance to the apoptosis-inhibiting effects of G-CSF and possibly GM-CSF, and appear more susceptible to the apoptosis-inducing action of TNFalpha, the greatest resistance being observed in the more neutropenic patients. PMID- 15895884 TI - IL1beta+3953 exon 5 and IL-2 -330 promoter polymorphisms in patients with rheumatoid arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA) is chronic inflammatory disease in which cytokines play an important role. The aim of present study was to evaluate the exon 5 +3953 IL-1beta and IL-2 -330 promoter polymorphisms in patients with RA in association with disease activity and severity. METHODS: In the study 93 patients with rheumatoid arthritis diagnosed according to the criteria of American College of Rheumatology were included. Polymerase chain reaction amplification was used for analysis of the polymorphisms studied. RESULTS: The distribution of IL-1beta and IL-2 genotypes in RA patients did not differ from control subjects. Nevertheless in patients with A2 allele of IL-1beta and GG genotype of IL-2, the active form of RA was more frequently diagnosed. Moreover in these patients the measurements of disease activity (DAS 28 score, ESR, number of swollen and tender joints) were significantly increased. CONCLUSION: We suggest that exon 5 +3953 IL1beta and IL-2 -330 promoter polymorphisms may be a genetic risk factor for RA severity. PMID- 15895885 TI - Increased levels of amino terminal propeptide of type III procollagen are an unfavourable predictor of survival in systemic sclerosis. AB - OBJECTIVE: Investigation of the impact on survival of inflammatory parameters (C reactive protein, ESR), markers of immune activation (serum soluble IL-2 receptor, soluble CD30), and N-terminal propeptide of type III procollagen levels (PIIINP) in systemic sclerosis (SSc). METHODS: In a prospective follow up study, clinical and laboratory data of 80 patients with SSc were evaluated. Kaplan-Meier survival curves and Cox proportional hazards model were used. Eighty cases with SSc were evaluated. Female/male ratio was 8/72. The mean (+/-SD) age was 49.3 (+/ 12.3) years, 16 patients died during our mean follow up of 58.1 months. RESULTS: In the univariate analysis, the presence of a C-reactive protein level above 20 mg/l was an unfavourable prognostic sign (p<0.001). Increased level of PIIINP level also caused an unfavourable outcome of disease (p<0.001). Conversely, increased ESR, soluble IL-2 receptor, soluble CD30 levels, presence of anaemia, did not influence the prognosis. Male gender (p<0.005), diffuse cutaneous SSc, clinically significant lung involvement (p<0.001), kidney (p<0.0001), cardiac (p<0.05) manifestations including pericarditis (p<0.02) were unfavourable prognostic signs by univariate Kaplan-Meier method. Multivariate analysis by Cox proportional hazards model showed that the increased level of PIIINP (RR: 6.98), and presence of diffuse cutaneous SSc (RR: 5.14) were independent unfavourable prognostic signs. CONCLUSIONS: An increased collagen metabolism unfavourably influences the outcome of SSc. This parameter may also be a potential candidate as a disease activity marker. PMID- 15895886 TI - Elevated whole blood chemiluminescence in patients with systemic sclerosis. AB - OBJECTIVE: Systemic sclerosis (SSc) is accompanied by oxidative stress that in turn may accelerate endothelium degeneration and thus disease progression. We tested whether phagocytes from SSc patients release more reactive oxygen species (ROS) and whether this release correlates with some clinical parameters. METHODS: ROS production by blood phagocytes was measured with the luminol enhanced whole blood chemiluminescence (CL). Resting and N-formyl-methionyl-leucyl-phenylalanine -induced CL (fMLP-induced CL) was measured in 30 patients with SSc and 30 healthy controls matched as to age, sex, and level of cigarette smoking. RESULTS: Resting CL and fMLP-induced CL calculated per 10(4) phagocytes present in the assayed blood sample were higher in patients with systemic sclerosis than in healthy controls (median; range, 0.88; 0.47-1.39 vs. 0.73; 0.13-1.07 aU/10(4)p and 621; 293-3522 vs. 411; 289-810 aUxs/10(4)p, p<0.02). Patients treated with cyclophosphamide and/or prednisone for 11; 3-168 months did not differ in respect to CL from those that never received the medications. Similarly, no significant differences were found between patients with limited and diffuse SSc. Resting CL correlated (p<0.05) with clinically manifested interstitial lung disease (r=0.59), single breath carbon monoxide diffusing capacity (r= -0.56) and serum autoantibodies titre (r= 0.43). CONCLUSIONS: Blood phagocytes from patients with systemic sclerosis, especially from those with interstitial lung disease, generate elevated amounts of ROS as assessed with CL. This confirms the presence of systemic oxidative stress in SSc patients. PMID- 15895887 TI - Prognostic factors of low bone mineral density in systemic sclerosis. AB - OBJECTIVE: To analyse the results of bone densitometry in patients with systemic sclerosis (SSc), evaluating the prognostic factors of low bone mineral density (BMD) in fertile and postmenopausal patients, and comparing to a control healthy group. METHODS: Cross-sectional study analysing 61 female SSc patients, aged 25 to 51 years, who performed a bone densitometry using dual x-ray absorptiometry. BMD values (lumbar spine, femoral neck, Ward and trochanter) infertile and postmenopausal patients were compared according to SSc clinical variant (limited and diffuse), race, previous use of drugs (corticosteroids and cyclophosphamide) and bone mass index (BMI). These results were compared with 47 fertile and 60 postmenopausal healthy women; multivariate linear regression analysis was used to study the influence of the variables of interest in the BMD results. RESULTS: Twenty-seven SSc patients presented osteopenia and 14 densitometric osteoporosis. No statistical association was found between BMD values and SSc clinical variants, race and previous use of corticosteroids and cyclophosphamide, in the fertile and in the postmenopausal groups. Fertile SSc patients were paired by age and race with the control group, but BMI (p = 0.035) was significantly lower in the SSc group. BMD values of lumbar spine (p = 0.070, statistical trend), femoral neck (p = 0.003), Ward (p < 0.001) and trochanter (p = 0.003) were significantly lower in the SSc group. Postmenopausal SSc patients were paired by age and race with the control group, but BMI (p < 0.001) was also significantly lower in the SSc group. Age at menopause (p = 0.006) was also significantly lower and time from menopause (p < 0.001) was significantly higher in the SSc group. BMD values of femoral neck (p < 0.001), Ward (p < 0.001) and trochanter (p = 0.001) were significantly lower in the SSc group. Multivariate linear regression analysis showed that BMI was the main variable influencing BMD in the fertile and postmenopausal groups. CONCLUSION: In the present study, BMD results in fertile and postmenopausal SSc patients were independent of the SSc clinical variants, race and previous use of corticosteroids and cyclophosphamide. A low BMD in appendicular sites was observed infertile and postmenopausal SSc patients when compared to a control healthy group, associated to a low BMI. PMID- 15895888 TI - Inflammatory cell infiltrate and RANKL/OPG expression in rheumatoid synovium: comparison with other inflammatory arthropathies and correlation with outcome. AB - OBJECTIVES: To evaluate if the immunofluorescence analysis of synovial tissue (ST) using antibodies against RANKL/OPG, conjugated with the immunophenotyping of lymphocytes and macrophages, could be of diagnostic and prognostic value in rheumatoid arthritis (RA) patients. METHODS: 3-year prospective study of 103 consecutive patients submitted to closed needle biopsy for diagnostic purposes. ST was analyzed with routine histologic techniques and immunofluorescence, using monoclonal antibodies against RANKL, OPG, CD163, CD68, CD4, CD8, interferon-gamma and CD19. Patients were prospectively evaluated with a clinical, laboratorial and radiological protocol. At the end of the follow-up patients were divided according to the final diagnosis. Results of the initial histologic evaluation were compared between the main diagnostic groups and in RA patients histologic data was correlated with clinical and radiologic outcome measures. RESULTS: The RANKL/OPG ratio and the inflammatory infiltrate were significatively higher in RA (n = 25) as compared to the same ratio observed in other inflammatory joint diseases (OIJD, n = 48) and in osteoarthritis (n = 17). The difference between RA and OIJD was specifically confirmed when the comparison involved spondyloarthropathy (n = 26). Final HAQ score and radiologic outcome were correlated with the density of intimal CD68+ macrophages. Radiologic progression was correlated with subintimal CD4+ lymphocytes and CD68+ macrophages and intimal CD68 and CD163+ macrophages. CONCLUSION: The quantification of the RANKL/OPG ratio and of the number of lymphocytes in the ST might be useful to differentiate RA from other inflammatory joint diseases. The ST number of CD4+ lymphocytes and macrophages are probable predictors of radiologic progression in RA patients. PMID- 15895889 TI - The development of bone mineral density and the occurrence of osteoporosis from 15 to 20 years of disease onset in patients with rheumatoid arthritis. AB - OBJECTIVE: To ascertain the occurrence of osteoporosis and the development of central bone mineral density (BMD) in long-term rheumatoid arthritis (RA) METHODS: BMD of the lumbar spine (L2-L4) and the femoral neck were measured by dual-energy X-ray absorptiometry in a cohort of 59 patients (49 women and 10 men) with rheumatoid factor-positive RA followed up for 20 years. BMD measurements were obtained at the 15- and 20-year follow-up visits. RESULTS: At the 15-year check-up the mean age was 61 (SD 13)for men and 54 (SD 11) years for women. Bone densitometry of these patients revealed decreased BMD at both lumbar spine and femoral neck, the mean T-scores being -1.1 [95%CI: -1.6 to -0.6] and -1.3 [95%CI: -1.6 to -1], respectively). Eighteen (31 %) patients thus had osteoporosis (BMD T -score < or = -2.5) and 32 (54%) patients were osteopenic (BMD T-score -1.0 to 2.5). However, when compared with reference values, the decreases in central bone mineral in this patient group were of low degree; the mean Z-score -0.2 [95%CI: 0.7 to 0.2] at the lumbar spine and -0.5 [95%CI: -0.8 to -0.3] at the femoral neck, respectively. After the subsequent five years the mean Z-score increased 0.45 [95%CI: 0.32 to 0.58] at the lumbar spine and the mean T-score decreased 0.20 [95%CI: -0.32 to -0.08] at the femoral neck. ESR, Larsen score, gender and cumulative dose of prednisolone during the 5 year follow-up and HAQ-index were used as explanatory parameters of BMD change between the 15- and 20-year follow ups. None of these parameters explained the BMD change. CONCLUSION: We conclude that in long-term RA central bone densities seemed to be only moderately decreased after 15 years from eruption of RA. No essential change in central BMD was found after the consecutive 5 years. PMID- 15895890 TI - Prevalence of antiphospholipid antibodies in systemic sclerosis and association with primitive pulmonary arterial hypertension and endothelial injury. AB - OBJECTIVE: To investigate the prevalence and clinical significance of antiphospholipid antibodies in patients with systemic sclerosis (SSc). METHODS: Autoantibodies against cardiolipin (aCL) and beta2-glycoprotein 1 (beta2-GPI) were detected by enzyme-linked immunoabsorbent assays (ELISAs) in successively hospitalised SSc patients admitted during a 24-month period. These patients were compared to patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA). RESULTS: 108 SSc patients were included: 61 had limited cutaneous SSc, 47 had the diffuse sub-type, 16 had primitive pulmonary arterial hypertension (PAH) and 34 had digital ulcerations. The control groups consisted of 37 RA and 38 SLE patients. The prevalence of aCL positivity was lower in SSc patients vs SLE patients (14 vs 47%; p < 0.001), lower in RA patients vs SLE patients (19 vs 47%; p < 0.001), and not different in SSc vs RA patients (14 vs 19%; NS). The mean aCL titer was also lower in SSc vs SLE patients (8+/-10 vs 15+/-20; p < 0.001). In SSc patients, positivity for aCL was associated with PAH (p = 0.009) and the aCL titer correlated with that of the von Willebrand antigen factor (r= 0.23; p = 0.045). The prevalence of anti beta2-GPI positive patients (IgG and/or IgM) was 5% in the SSc group, 18% in the SLE group and 5% in the RA group (SLE vs SSc and SLE vs RA; p = 0.005). CONCLUSION: We found that the prevalence of antiphospholipid antibodies in SSc patients was low. However, aCL antibodies were associated with PAH and endothelial injury. PMID- 15895891 TI - Altered thiol pattern in plasma of subjects affected by rheumatoid arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA) is a chronic inflammatory disease which involves the synovial membrane of multiple diarthroidal joints causing damage to cartilage and bones. The damage process seems to be related to an overproduction of oxygen reactive species inducing an oxidative perturbation. Since sulfhydryl groups are primary antioxidant factors, we were interested in investigating the balance of plasma sulfhydryl/disulfides in patients with active RA compared to healthy control subjects. METHODS: Twenty-one patients with RA and 15 age-matched controls were studied. Plasmatic sulfhydryl groups and their disulfide form concentrations were measured by spectrophotometry or HPLC. RESULTS: RA patients showed significantly lower levels of plasma protein sulfhydryls and cysteinyl glycine compared to healthy controls (p < 0.001). Conversely, cystine and homocystine, and protein-bound cysteine and homocysteine were significantly increased (p < 0.005 in disulfides forms and p < 0.05 in protein mixed disulfides forms). There was a significant correlation between some clinical data (ESR, number of tender/swollen joints) and some of the parameters studied. CONCLUSION: The results of this study indicate a biochemical disturbance of plasma sulfhydryl/disulfides balance in patients with RA compared to controls with an increase in some oxidised forms (disulfides and protein mixed disulfides) and a decrease in free thiols. The increase in total homocysteine, correlated to the higher risk of cardiovascular diseases in RA patients, is associated with higher levels of the oxidised forms, disulfides and protein-thiol mixed disulfides. PMID- 15895892 TI - Comparison of two hyaluronan drugs in patients with advanced osteoarthritis of the knee. A prospective, randomized, double-blind study with long term follow-up. AB - OBJECTIVES: To compare the long-term effects of high and low molecular weight hyaluronic acid (HA) applications in severe (Kellgren Lawrence stage III) osteoarthritis (OA) of the knee. METHODS: In a prospective clinical trial 184 knees (92 patients) with radiographic Kellgren Lawrence stage III OA were randomized to receive either 3 intra-articular high molecular weight HA (Hylan G F 20) injections or 3 low molecular weight HA (Orthovisc) injections at one-week intervals. Patients were evaluated by the Hospital for Special Surgery (HSS) Knee Score and were followed-up for 12 months. RESULTS: The total HSS score in high molecular weight HA patients improved from 71.8+/-11.6 to 86.7+/-11.6 and in low molecular weight HA patients from 66.7+/-11.0 to 86.6+/-9.1 at the end of the trial (p < 0.01). There were no statistically significant differences between the groups and both had improved in all parameters at the latest follow-up (p = 0.000). CONCLUSIONS: Three intra-articular injections at intervals of 1 week of both HA preparations resulted in a pronounced reduction in pain and improved function as measured by the HSS score during a period of 52 weeks, without complications. PMID- 15895893 TI - Cytokine production in thromboangiitis obliterans patients: new evidence for an immune-mediated inflammatory disorder. AB - OBJECTIVE: To study IL-6, IL-10, IL-12 levels and circulating immune complexes (CIC) containing IgG, IgM or IgA in sera of 14 TAO patients and 12 healthy blood donors. To evaluates the ability of TAO PBMC to produce IL-6, IL-10, IL-12, as well as to detect PBMC apoptosis after stimulation with different stimuli. METHODS: In vitro stimulation of PBMC with lypopolysaccharide (LPS), phytohemagglutinin (PHA), C3 binding glycoprotein from uscuta europea (C3bgp), pokeweed mitogen (PWM), and dexamethasone (DM) were performed. The quantities of the secreted cytokines in sera and in culture supernatants, as well as CIC were detected by ELISA. The apoptosis was assessed according to nuclear morphology, after acridine orange staining, by fluorescence microscopy. RESULTS: Significantly higher IL-6 levels in the patients', than in the controls' sera was found. An increased production of IL-6 and IL-12 in TAO PBMC supernatants was detected, regardless of the stimuli used. A hyporeactivity of TAO PBMC toward IL 10 production was found after C3bgp, LPS, PHA and PWM stimulation, compared to the controls' PBMC. The spontaneous and induced apoptosis was significantly higher in TAO compared to the control group. Increased CIC quantities were detected in 75% of the patients tested. According to the CIC isotype, the IgG CIC positives (75%) prevailed over IgA CIC positives (50 %). CONCLUSION: The altered production of IL-6, IL-12 and IL-10, the increased apoptosis as well as the elevated levels of CIC could be a reason for the persisting immune inflammation in TAO. PMID- 15895894 TI - In vitro production of myeloperoxidase anti-neutrophil cytoplasmic antibody and establishment of Th1-type T cell lines from peripheral blood lymphocytes of patients. AB - OBJECTIVE: To investigate the pathogenic role of T cells in the development of anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitis. METHODS: Peripheral blood lymphocytes (PBL) were isolated from myeloperoxidase anti neutrophil cytoplasmic antibody (MPO-ANCA) associated vasculitis patients and cultured in medium. The production of MPO-ANCA in the medium of PBL stimulated with Concanavalin-A (Con-A), with or without cyclosporin (CyA), was measured by enzyme-linked immunosorbent assay (ELISA) on MPO coated plates. RNA isolated from PBMC of one patient was used for polymerase chain reaction (PCR) and single stranded conformational polymorphism (SSCP) studies, and MPO-specific T cell lines (TCL) were established by antigen stimulation techniques. RESULTS: PBL of patients with MPO-ANCA-associated vasculitis produced MPO-ANCA following Con-A stimulation, and this effect was inhibited by treatment with cyclosporin A (CyA) or elimination of CD4 cells. PCR-SSCP showed autoantigen-reactive oligoclonal T cell accumulation in PBMC of one of these patients. We established MPO-specific TCL which secreted interferon-gamma (IFN-gamma), but not interleukin-4 (IL-4); all TCL were CD4 positive, CD8 negative, and HLA-DR restricted. CONCLUSIONS: Our results suggest that Th1-type T cells may mediate MPO-ANCA production, and may play a role in the onset of MPO-ANCA vasculitis. PMID- 15895895 TI - Post-treatment residual tissue in idiopathic retroperitoneal fibrosis: active residual disease or silent "scar" ? A study using 18F-fluorodeoxyglucose positron emission tomography. AB - OBJECTIVE: Medical treatment is often effective in idiopathic retroperitoneal fibrosis (IRF) but frequently leads to residual retroperitoneal masses that may represent active disease or simply consist of inactive fibrotic tissue. 18F fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) is a functional imaging modality that reliably assesses disease activity in a number of inflammatory diseases including IRF. We used 18F-FDG PET to evaluate the metabolic activity of residual masses in a series of IRF patients. METHODS: We studied 7 consecutive IRF patients, all of whom presented constitutional symptoms and/or pain, and had high acute-phase reactant levels; 6 had ureteral involvement. IRF was diagnosed by means of computed tomography (CT), which revealed a peri-aortoiliac mass in all cases. Three patients underwent surgical ureterolysis and 2 received ureteral stents. Subsequently, 5 patients received prednisone, one sequential treatment with prednisone and tamoxifen, and one prednisolone plus methotrexate. All of the patients underwent 18F-FDG PET at varying times after the end of treatment. RESULTS: The presenting signs/symptoms improved in all patients and the levels of acute-phase reactants significantly decreased or normalised. Ureteral obstructive disease resolved in all cases. Post treatment CT revealed a considerable reduction in the amount of IRF but all of the patients had a residual retroperitoneal mass. PET revealed slight aorto-iliac 18F-FDG uptake in only one patient; all of the others were negative. No patient relapsed during the follow-up. CONCLUSIONS: Post-treatment residual masses are frequent in IRF patients but, in most cases, probably represent metabolically inactive tissue. PMID- 15895896 TI - How to define a Minimal Clinically Individual State (MCIS) with pain VAS in daily practice for patients suffering from musculoskeletal disorders. AB - OBJECTIVES: Pain is frequently the primary variable in symptomatic clinical trials for the evaluation of rheumatological disorders. The protocol of such trials mention a minimum level of pain as an entry criterion [e.g. a level above the Patient Acceptable Symptoms State (PASS)] and the changes in pain as the primary variable. Usually, the results are expressed at a group level as the mean changes in pain. However, the presentation at an individual level and, in particular, the percentage of patients with a Low Disease Activity State at the end of the study seems more clinically relevant. Pain is usually evaluated using a continuous variable such as a 0-100 visual analogue scale. The cut-offs permitting one to define both the entry criterion and the LDAS are not well established. The objective of this study was to evaluate such cut-offs using a patient-derived perspective. METHODS: STUDY DESIGN: cross-sectional study. PATIENTS: consecutive out patients suffering from chronic rheumatic diseases familiar with the use of a VAS to evaluate their level of pain. DATA COLLECTED: two questions were asked the patients at the end of the visit: "Based on the experience you have because of your chronic rheumatic disorder, could you please specify the level of pain below which you consider your disease as inactive ? Moreover, could you please also specify the level of pain above which you consider taking a pain killer?" Before answering the second question, it was explained to the patient that their answer to the second question could be similar to their response to the first one. For the two questions, the cumulative percentage of patients (disease inactive and pain killer intake) were calculated for each level of pain. RESULTS: The underlying disease of the 137 evaluated patients (mean age: 57+/-16 and female sex: 76%) was rheumatoid arthritis (n = 59), ankylosing spondylitis (n = 19), SLE (n = 2), back pain (n = 20), or peripheral osteoarthritis (n = 37). The mean disease duration was 12+/-10 years. At the time of the study, the current level of pain evaluated on a 0-100 VAS was 33+/-22. The LDAS was 49, 36 and 25 for our patient population at the 25th, 50th and 75th percentiles, respectively. The pain killer intake level was 32, 48, 64 at the 25th, 50th, 75th percentile respectively. CONCLUSION: This study suggests that LDAS and PASS may be distinct concepts. The methodological approach adopted here could be of interest for specifying the minimum level of symptoms at entry in a symptomatic trial (PASS) and also to present results in terms of the percentage of patients in good condition (LDAS) at the end of a trial. PMID- 15895897 TI - Monocyte chemoattractant protein-1 activates a regional Th1 immunoresponse in nephritis of MRL/lpr mice. AB - OBJECTIVE: Monocyte chemoattractant protein-1 (MCP-1) is upregulated and recruits and activates inflammatory cells in nephritis of MRL lpr mice. It has been shown that anti-MCP-1 gene therapy is specifically effective in nephritis, while it was apparent that an imbalance towards Th1 predominance accelerates nephritis in MRL/lpr mice. The aim of this study was to clarify whether blockade of the MCP-1 signal by anti-MCP-1 gene therapy influences the Th1/Th2 balance in MRL/lpr mice. METHOD: An NH2-terminal deletion mutant of the MCP-1 gene (7ND) was injected into the skeletal muscles of MRL/Ipr mice with advanced stage nephritis to suppress MCP-1 and its receptor (CCR2) signaling pathway. We evaluated the local tissue production of cytokines in splenocytes and microdissected infiltrating cells within the glomeruli or interstitium. RESULT: Although the production of cytokines in splenocytes was not influenced by anti-MCP-1 gene therapy, kidney glomeruli IL-12 mRNA production and interstitium-infiltrating cell production of IL-12 and IFN-gamma mRNA were significantly reduced. CONCLUSION: The blockade of MCP-1 gene therapy does not influence helper T cell polarization, but acts directly on the regional Th1 immunoreaction in MRL/lpr mice. PMID- 15895898 TI - Serum soluble interleukin-2 receptor predicts early remission in patients with recent-onset rheumatoid arthritis treated with a single disease-modifying antirheumatic drug. AB - OBJECTIVE: To study the value of baseline serum levels of circulating soluble interleukin-2 receptor (sIL-2R) and soluble E-selectin as predictors of early remission in patients with recent-onset rheumatoid arthritis (RA) receiving a single disease-modifying anti-rheumatic drug (DMARD) (SINGLE) or therapy with a combination of DMARDs (COMBI). METHODS: Baseline (n = 157) serum samples originate from the FIN-RACo (FINnish Rheumatoid Arthritis Combination therapy) trial, in which 195 patients with early and clinically active RA were randomly assigned to receive either SINGLE (initially sulfasalazine) with or without prednisolone, or COMBI therapy (sulfasalazine, methotrexate, hydroxychloroquine, and prednisolone). Of the samples, 76 were from SINGLE patients and 81 from COMBI patients. sIL-2R was measured by automated immunoassay analyzer and sE-selectin by enzyme-linked immunosorbent assay. RESULTS: At six months, 7 (9% [95% CI: 4 to 18]) SINGLE and 19 (23% [95% CI: 15 to 34]) COMBI patients were in remission. In multivariate logistic regression analysis, sIL-2R <442 U/ml and COMBI therapy were the only predictors of remission. The area under receiver operating characteristic curve for sIL-2R level was 0.86 (95% CI: 0.62 to 0.95) in SINGLE and 0.57 (95% CI: 0.42 to 0.71) in COMBI (p = 0.006). In SINGLE, the optimal cut offpoint was 442 U/ml, lower levels predicting remission with sensitivity of 83% (95% CI: 73% to 91%) and specificity of 86% (95% CI: 42% to 100%). Likelihood ratio for positive test was 5.9 (95% CI: 1.6 to 32.8). In multivariate logistic regression analysis, sIL-2R <442 U/ml and COMBI therapy were the only predictors of remission. CONCLUSION: Low baseline serum sIL-2R level predicts early remission of patients with active early RA treated with a single DMARD. PMID- 15895899 TI - Drug-induced agranulocytosis during treatment with infliximab in enteropathic spondyloarthropathy. AB - Agranulocytosis is a disorder characterized by a severe decrease in the number of granulocytes in blood, that frequently occurs as an adverse reaction to some drugs. By now, there are no reports in literature of agranulocytosis caused by tumur necrosis factor-alpha blockers. We describe the case of a 20-year-old Caucasian male affected by enteropathic (Crohn's disease) spondyloarthropathy HLA B27 negative, successfully treated with infliximab. After the second infliximab infusion, he was found to have a severe transient neutropenia (0.5 x 10(9)/L). Routine serum chemistry and full blood cell count (apart from neutrophil count) were normal. Serology excluded an active infection. Bone marrow needle aspirate showed a normal trilineage differentiation. Autoantibody assessment showed negative ANA, anti-dsDNA, anti-ENA, and ANCA, but positive granulocyte-bound antibodies (GBA) and neutrophil-specific (CD 16+)-bound antibodies (anti-NA). Ten weeks after infliximab infusion, neutrophil count and GBA and anti-NA assay returned spontaneously within normal range and we observed the same progress after every successive infliximab infusion we performed. These data indicated that infliximab possibly triggered production of granulocyte and neutrophil autoantibodies with resultant autoimmune agranulocytosis. PMID- 15895900 TI - Pseudopodagra: A presenting manifestation of infective endocarditis. AB - Podagra is a term used to describe acute monoarthritis of the first metatarsophalangeal (1st MTP) joint. The most common diagnoses of arthritis in this joint are: crystal-induced synovitis, septic arthritis, traumatic conditions and reactive arthritis. When etiologies other than gout are involved this is frequently referred to as pseudopodagra. We report the case of a patient who presented with pain and swelling of the 1st MTP The absence of intraarticular crystals and hyperuricemia encouraged further evaluation of the patient. A cardiac murmur was investigated by echocardiography, which revealed valvular vegetations and the diagnosis of infective endocarditis (IE) was established. This is the first reported case of a podagra-like presentation of IE. As in this case, the diagnosis of gout should rest on findings beyond the presence at 1st MTP arthritis, with evaluation of all extraarticular signs in order to rule out other possible diagnoses. PMID- 15895901 TI - The future of the treatment of systemic lupus erythematosus. AB - Despite recent advances, patients with systemic lupus erythematosus (SLE) still experience considerable morbidity and mortality. To try and improve their prognosis, varied novel biological interventions and immune manipulations are being developed. They may hold promise in particular for patients whose disease is organ-threatening and refractory to conventional treatment. In addition, awareness of the tendency of lupus patients to develop accelerated atherosclerosis as well as newly gained insights into the underlying mechanisms, may lead to better control of risk factors, earlier diagnosis of prevalent cardiovascular disease and more effective treatment. Infections also remain a significant threat that may be amenable to improved preventive measures. Evidence related to a better management of lupus patients by specialists, the need to address the impact of commonly associated stress and depression and other significant developments are also presented and discussed. PMID- 15895902 TI - Disease duration, hypertension and medication requirements are associated with organ damage in childhood-onset systemic lupus erythematosus. AB - OBJECTIVE: To investigate the frequency of organ damage in childhood-onset systemic lupus erythematosus (SLE) and to identify disease variables and patient characteristics related to organ damage. METHODS: A cohort of 71 patients was examined in a cross-sectional study after a mean disease duration of 10.8+/-8.2 years (mean age 26.4+/-9.8 years). The occurrence of organ damage was measured by the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SDI). Factors analysed as possible explanatory variables of organ damage were the following: demographic variables, clinical variables at diagnosis and during disease course, as well as medication use. Growth and self-reported health status were also measured. RESULTS: The most frequent areas of organ damage were in the neuropsychiatric (28%), renal (13%) and musculoskeletal (13%) organ systems. Forty-three patients (61%) had evidence of damage. The mean SDI score was 1.3 for the whole study population. Hypertension, longer disease duration and use of cyclophosphamide were factors significantly related to an increasing SDI score in multiple linear regression analyses. Furthermore, patients with damage (SDI > or =1) compared to those without damage (SDI = 0) had a significantly higher cumulative corticosteroid dose (24.7 g versus 10.6 g) and more frequently required high-dose prednisolone at diagnosis (68% versus 43%). CONCLUSION: Evidence of organ damage was found in 61% of all patients. Long disease duration, known hypertension and use of cylophosphamide were significantly associated with an increasing SDI score. Furthermore high-dose prednisolone at diagnosis and cumulative prednisolone dose were significantly related to the presence of organ damage. PMID- 15895903 TI - Stapling of knees with valgus deformity in children with juvenile chronic arthritis. AB - OBJECTIVE: Leg length discrepancy and excessive knee valgus are potential complications of juvenile chronic arthritis of the knee. The aim of the present study was to evaluate retrospectively the safety and efficacy of temporary stapling of the knee epiphyses in management of valgus deformities of the knee in children with JCA. METHODS: Medical data of the patients with temporary epiphyseodesis due to knee valgus deformity (KVD) were studied. 177 knees in 112 patients were found with sufficient data for evaluation. Patient documents and radiographs of these patients were evaluated. RESULTS: Mean age at the time of operation was 8 years (range: 2 - 17) in 19 males and 93 females. The patients are predominantly affected by aggressive polyarticular disease. Preoperative mean valgus angle was 11 degrees (IQR: 9, 14) and at staple removal 4 degrees (IQR: 2, 5). In 120 of 177 knees (68% [95% CI: 61 - 74], p < 0.001) the physiological angle (3-8 degrees) was reached. Median time of stapling was 10 months (IQR: 8, 13). Five reversible and one irreversible (3% [95% CI: 2 to 7]) major complications were documented among the 177 stapled knees. CONCLUSION: Temporary epiphyseal stapling enables flexible correction of KVD in children with JCA. Low complication rate encourages the use of the method. Prompt follow-up is, however, important in avoing excess over-correction to varus. PMID- 15895904 TI - Lupus erythematosus manifestations exacerbated by etanercept therapy in a patient with mixed connective tissue disease. PMID- 15895905 TI - Do MEFV mutations play a role in the development of Behcet's disease related amyloidosis? PMID- 15895906 TI - New-onset acute heart failure and ventricular tachycardia after therapy with a tumor necrosis factor antagonist. PMID- 15895907 TI - Chondrosarcoma in Paget's disease of bone. PMID- 15895908 TI - Fibrotic involution of sural muscles secondary to lupus panniculitis. PMID- 15895909 TI - Thyroid function, autoimmune thyroiditis and coeliac disease in juvenile connective tissue diseases. PMID- 15895910 TI - Systemic lupus erythematosus in a patient with beta-thalassemia major. PMID- 15895911 TI - Seronegative spondyloarthropathy associated with Takayasu's arteritis in a child. PMID- 15895912 TI - Myocarditis in a girl as an initial manifestation of systemic lupus erythematosus. PMID- 15895913 TI - Psychiatric rehabilitation and the science of possibilities. PMID- 15895914 TI - Moving forward: negotiating self and external circumstances in recovery. AB - This article presents a framework for understanding the concept of recovery from serious mental illnesses and other life struggles. The framework is based on findings from a longitudinal, qualitative study that involved in-depth interviews with 28 people who experienced serious mental health challenges. The purpose of this article is to clarify the concept of recovery by presenting a grounded theory analysis of the components of recovery. The framework recognizes the experiences of struggle constructed through the words of study participants and captures four main components of recovery: a) a drive to move forward, b) a spiral of positive and negative changes, c) the context of recovery, and d) a dialectical process of ongoing negotiation between self and external circumstances. PMID- 15895915 TI - Consumer-run drop-in centers: program operations and costs. AB - In-depth phone surveys were conducted with 32 consumer-run drop-in centers in Michigan. Results indicate that centers serve a diverse array of consumers at an average cost of about dollar 8 daily per person. Funding levels, salaries, and services are quite heterogeneous among centers. Those with higher funding levels, greater involvement with other human service agencies, and higher overall CMH county budgets differed significantly in total services and activities provided than those centers with less of each of these resources. Daily attendance was predicted by other-agency involvement, participation of volunteer personnel, and negative neighborhood context. PMID- 15895916 TI - Supported employment for people with co-occurring disorders. AB - Research shows that people with dual disorders (i.e., a co-occurring mental illness and substance use disorder) are successful in supported employment programs and that employment can be a crucial step in their recovery. Based on experience observing supported employment services for 15 years, we propose practice guidelines for people with dual disorders. Successful programs share several approaches: 1) encourage employment, 2) understand substance abuse as part of the vocational profile, 3) find a job that supports recovery, 4) help with money management, and 5) use a team approach to integrate mental health, substance abuse, and vocational services. PMID- 15895917 TI - Primary care experiences of people with psychiatric disabilities: barriers to care and potential solutions. AB - This study examined the barriers that 16 focus group participants with psychiatric disabilities confront in obtaining primary care services and their recommendations on improving quality of care. They cited a) difficulty identifying a primary care physician with good empathic and communication skills, b) physicians' misunderstanding of the nature of psychiatric disability, c) inadequate information about the side effects of psychotropic medications, and d) costs due to inadequate insurance coverage. In addition to suggestions for improving patient-physician communication and expanding physician knowledge, participants emphasized strategies to become empowered in their relationships with physicians and to obtain personal support. PMID- 15895918 TI - Identifying the core competencies of community support providers working with people with psychiatric disabilities. AB - The study was intended to identify core competencies for community support providers working with people with psychiatric disabilities. Using multiple methods developed from previous research in the field of developmental disabilities, 18 consumers receiving services and 16 staff members from two mental health community support programs identified a list of 68 competencies that included personal attributes, knowledge, and skills. Based on a card sort task, 34 consumers receiving services and 34 support workers from six mental health community support programs rated 59 of the 68 competencies as being either absolutely necessary or desirable. Results of a second card sort task found that a majority of competencies identified as being needed pre-employment were personal attributes consistent with adopting a person-centered approach. Competencies categorized as to be learned on the job involved special knowledge and skills specific to working with people with psychiatric disabilities. The range of personal attributes, knowledge, skills represented in the identified competencies reflects the complexity of contemporary mental health community support. Findings are indicative of the need for specialized training and supervision that has not been typically available in the community mental health sector. PMID- 15895919 TI - What affects self-esteem of persons with psychiatric disabilities: the role of causal attributions of mental illnesses. AB - This study addresses the factors which affect the self-esteem of persons with psychiatric disabilities, with a specific focus on the role of causal attributions of mental illnesses. It is based on data collected from 461 persons with psychiatric disabilities (consumers) served through psychiatric rehabilitation agencies. The results of regression analyses revealed that household income, diagnosis, psychiatric symptoms, service satisfaction, perceived stigma, and perceptions of social roles were all related to self esteem. Consumers' causal attributions did not have direct effects on self esteem, but the effects of causal attributions on self-esteem were moderated by psychiatric symptoms and by perceptions of social roles, suggesting that self esteem enhancement strategies should be tailored to consumers' psychiatric and cognitive characteristics. PMID- 15895920 TI - A strengths perspective in working with an adolescent with depression. AB - Depression has long been a problem in our society. In this paper, the writer tries to contrast a disease orientation and a strengths perspective in helping an adolescent with depression. The strengths perspective advocates identifying needs and ability behind client's symptoms and deficits; nurturing individuals' positive mood and hope to recovery; establishing enlightening encounter between worker and client; and developing positive coping in working with individuals with depression. PMID- 15895921 TI - Factors influencing success in a Fairweather Model mental health program. AB - A mixed design study explored factors contributing to success and satisfaction in a Fairweather Model mental health organization. Methodology included chart audits, employee surveys, and resident interviews. The study found multiple factors leading to success in this model with emphasis on motivation, mental and chemical health, and family support. Comprehensive services, high quality of living, and peer support were identified as leading to success and satisfaction in the model. PMID- 15895922 TI - From rhetoric to routine: assessing perceptions of recovery-oriented practices in a state mental health and addiction system. AB - The Recovery Self Assessment (RSA) was developed to gauge perceptions of the degree to which programs implement recovery-oriented practices. Nine hundred and sixty-seven directors, providers, persons in recovery, and significant others from 78 mental health and addiction programs completed the instrument. Factor analysis revealed five factors: Life Goals, Involvement, Diversity of Treatment Options, Choice, and Individually-Tailored Services. Agencies were rated highest on items related to helping people explore their interests and lowest on items regarding service user involvement in services. The RSA is a useful, self reflective tool to identify strengths and areas for improvement as agencies strive to offer recovery-oriented care. PMID- 15895923 TI - Weight gain information on the Internet for people who have schizophrenia. AB - Weight gain and chronic health problems are highly prevalent among people with schizophrenia. Basic information about diet and exercise changes would be very useful to people with schizophrenia as well as the family members and clinicians. Because people increasingly seek information from the World Wide Web, this paper reports on a survey of weight-related information on available web sites. A survey of 21 schizophrenia websites revealed that although fourteen sites (67%) included some information about weight gain and related health problems, more than half of the information was limited to site user-generated comments, and consisted of less than one paragraph of information. PMID- 15895924 TI - Role development: an evidenced-based intervention for individuals diagnosed with schizophrenia in a forensic facility. PMID- 15895925 TI - The client experience of assertive community treatment: a qualitative study. AB - Assertive Community Treatment (ACT) has demonstrated effectiveness in reducing hospital days for persons with schizophrenia. Utilizing depth interviews and Grounded Theory techniques (Strauss & Corbin, 1998), client reactions to this intensive form of treatment delivery were explored. This study illustrates the importance of the relationship between treatment providers and persons with schizophrenia. As providers assisted clients with practical needs, clients began to develop trust, which fostered a sense of belonging and relationship, leading toward a positive motivation regarding treatment. PMID- 15895926 TI - The effects of yoga on mood in psychiatric inpatients. AB - The effects of yoga on mood were examined in 13 psychiatric inpatients at New Hampshire Hospital. Participants completed the Profile of Mood States (POMS) prior to and following participation in a yoga class. Analyses indicated that participants reported significant improvements on all five of the negative emotion factors on the POMS, including tension-anxiety, depression-dejection, anger-hostility, fatigue-inertia, and confusion-bewilderment. There was no significant change on the sixth POMS factor, vigor-activity. Improvements in mood were not related to gender or diagnosis. The results suggest that yoga was associated with improved mood, and may be a useful way of reducing stress during inpatient psychiatric treatment. PMID- 15895927 TI - Enriching the process of "good science" through participation of people with psychiatric disabilities in all aspects of research. AB - The purpose of this study is to describe the experiences of researchers with a psychiatric disability participating in a project evaluating the services of Community Mental Health Centers (CMHCs) and to identify strategies for researchers with a psychiatric disability in mental health. A project coordinator and four researchers who were people with psychiatric disabilities were interviewed. The rationale for researchers with a psychiatric disability is discussed, and the facilitators of and barriers to them are described. People with psychiatric disabilities should be involved in formulating research questions, data collection, interpreting the results, identifying implications, and presenting the results to stakeholders. People with psychiatric disabilities' participation in CMHC program evaluation can promote service innovation and enhance people with psychiatric disabilities'satisfaction. PMID- 15895928 TI - [Febrile state without clinically evident signs: does it mean sepsis? Can one make a rapid determination?]. PMID- 15895929 TI - [Usefulness of procalcitonin in adults and children]. AB - In febrile children and adults, it is frequently difficult, based on the sole clinical examination, to differentiate a bacterial illness from systemic inflammatory syndromes or severe viral infections. However, the positive and rapid diagnosis of a severe bacterial infection or a sepsis is essential to initiate lifesaving therapies. Among the numerous infectious biomarkers that have recently been investigated, procalcitonin has the best diagnostic yield. Plasma levels below 0.5 microg/l usually rule out a severe bacterial disease, whereas values above 2 microg/l are strongly indicative of a bacterial sepsis. The usefulness and the limitations of the measurement of procalcitonin as a diagnostic and a prognostic tool during severe bacterial infections are discussed in this paper. PMID- 15895930 TI - [Utility of procalcitonin for the diagnosis and the follow-up of infections in febrile neutropenic patients]. AB - Procalcitonin is a marker of severe bacterial infections in non-neutropenic patients. The goal of this review is to assess its utility in the management of neutropenic patients. A delayed treatment of infection in this setting results in severe morbidity and high mortality. As traditional diagnostic tools often fail to exclude infection when fever occurs, all these patients receive empirical antimicrobial therapies during long periods of time. Present knowledge suggests that procalcitonin may contribute to identify patients in whom 1) antibiotics could be stopped in the absence of bacterial infection, 2) investigations and adjustments of the antimicrobial therapy for persistent fever are needed. The use of procalcitonin for the management of febrile neutropenic patients should be studied prospectively. PMID- 15895931 TI - [Detection and quantification of nucleic acids in infections diseases: utility and limitations]. AB - The amplification of nucleic acids is often used for the diagnosis and the follow up of infectious diseases. The interpretation of PCR results varies according to the pathogens detected, the site of infection and the clinical presentation. PCR tests might be the reference method or only an help for the diagnosis. With the development of antiviral treatments quantitative PCR tests are now essential for the evaluation of treatment efficacy. A discussion with the laboratory is important for the correct interpretation of PCR results. PMID- 15895932 TI - [Chlamydia: diagnostic and treatment]. AB - Chlamydia are obligate intracellular bacteria. Three species are considered human pathogens. Chlamydophila pneumoniae is one of the most common agents of atypical community-acquired pneumonia. Chlamydophila psittaci causes psittacosis, a severe zoonotic pneumonia transmitted by birds. Finally, Chlamydia trachomatis is the etiologic agent of trachoma and urogenital infections. The latter are commonly asymptomatic or paucisymptomatic. Thus, they may remain undiagnosed for years, leading to serious late complications such as salpingitis, ectopic pregnancy and infertility. Currently, the diagnosis of chlamydial infections is essentially based on molecular methods. Treatment should use an antibiotic with good intracellular bioavailability such as tetracycline, macrolides and new generation fluoroquinolones. PMID- 15895933 TI - [Diagnosis of invasive fungal infections]. AB - The diagnosis of invasive fungal infections--in particular those caused by Candida and Aspergillus--is difficult and often delayed. The conventional laboratory diagnosis relies on direct microscopic examination and culture of relevant clinical material, as well as on histology. To improve and speed up the microbiological diagnosis, two new approaches have been thought, the detection of circulating cell wall fungal antigens (galactomannane from Aspergillus, mannane from Candida, glucane), and the detection of fungal specific nucleic acids by PCR directly in clinical materials. Among these still experimental methods, the serial detection of galactomannane in the serum of at risk patients is now considered as a useful adjunct for the early diagnosis of invasive aspergillosis. PMID- 15895934 TI - [Hemolytic uremic syndrome in childhood: not so rare in Swiss children]. AB - Fifty years after the first report by Gasser and Gautier, hemolytic, uremic syndrome is rather rare but severe childhood disease. A recent survey demonstrates that more than 90% of the cases occurring in Switzerland are caused either by Escherichia coli that produces shigatoxin or by Streptococcus pneumoniae. PMID- 15895935 TI - [Psychotropic medication traps in children and adolescents]. PMID- 15895936 TI - [What is a good physician?]. PMID- 15895937 TI - [The loss of the "memory of water"]. PMID- 15895938 TI - [Colon cancer: a new French medical affair?]. PMID- 15895939 TI - [Life? health? water!]. PMID- 15895940 TI - Non-immune hydrops fetalis: prenatal diagnosis and perinatal outcomes. AB - PURPOSE: To describe the perinatal outcomes after a prenatal diagnosis of non immune hydrops fetalis (NIHF). METHODS: Retrospective case series. University of Mississippi ultrasound database from July 2000 to January 2003 was reviewed for cases of NIHF. Records abstracted for maternal demographics and neonatal outcomes. RESULTS: Thirty-one cases of NIHF were identified and full data was available for 29 cases. Median gestational age at diagnosis was 17.3 weeks. Structural fetal malformations were noted in 22 cases (76%); most common were cystic hygroma (N = 17, 59%), followed by cardiac (N = 10, 34%). Amniocentesis was performed in 20 cases and 10 were aneuploid. Only 10 fetuses (34%) were liveborn and only five (17%) survived the neonatal period. CONCLUSIONS: Antepartum diagnosis of NIHF conveys a grave perinatal prognosis. Associated anomalies can be expected in approximately 75% of cases. Even for those fetuses achieving viability, survival beyond the neonatal period is rare. PMID- 15895941 TI - Clinical problem-solving: shock and ahhh. PMID- 15895942 TI - Health savings accounts. PMID- 15895943 TI - Not "whether" but "how". PMID- 15895944 TI - The influenza epidemic at the Normal College in 1918. 1919. PMID- 15895945 TI - The unseen aesthetics of double decker televisions: a Southernism. PMID- 15895946 TI - [Bipolar disorders: improvement in their screening and management]. PMID- 15895947 TI - [Lyme borreliosis]. AB - Lyme borreliosis is a bacterial tick borne infection due to Borrelia burgdorferi sensu lato. In 50% of the cases, the disease begins by a centrifuge erythema migrans localized around the tick bite. Neurological, articular and rarely cardiac, ocular or cutaneous manifestation may be observed as secondary manifestations. They sign the dissemination, the proliferation of Borrelia burgdorferisensu lato and the development of the immune response. In the absence of erythema migrans, secondary clinical manifestations may be difficult to diagnose. Direct detection of Borrelia burgdorferi sensu lato by culture is fastidious, time consuming and of a low sensitivity. Molecular methods (polymerase chain reaction) are very specific but limited by a low bacterial burden in the tissues. Serological testing interpretation is tricky but guides the diagnosis at the disseminate phase. Curative antibiotherapy is efficient, especially at the initial phase of infection. The two main elements for prophylaxis are individual protection from tick bites and a rapid and complete tick extraction. Antibioprophylaxy of tick bites is not recommended and a strict clinical surveillance must be done. PMID- 15895948 TI - [Bipolar disorders: frequent but difficult and delayed diagnosis]. PMID- 15895949 TI - [Epidemiology of bipolar disorders]. AB - Recent emerging data provide converging evidence for a high prevalence of bipolar disorders (up to 5% of the general population). The proper recognition of the entire clinical spectrum of bipolarity, including subsyndromal manic/hypomanic symptoms is of major public health importance. Indeed, accurate diagnosis and appropriate treatment still lag by many years, leading to greater psychosocial impairments and higher suicidality. Earlier risk factor assessment could therefore substantially decrease the occurrence of suicidal behaviors. PMID- 15895950 TI - [Pathogenesis of bipolar disorders: genetic vulnerability and environmental factors]. AB - It is now well-established that there is a genetic vulnerability for developing bipolar disorders. For instance, the rate of bipolar disorders in first degree relatives of bipolar probands (19%) is higher than in the general population (1%); monozygote twins have higher concordance rates for bipolar diseases (67%) than dizygote (19%); finally affective disorders are more frequent in biological parents than in adoptive parents in adopted bipolar patients. However, currently genes predisposing for the disease are not known, in part because bipolar disorders are very heterogeneous. New research strategies should be developed based on more homogeneous groups of patients defined by the presence of "candidate symptoms" or by the evidence of endophenotypes in healthy relatives. However, it is clear now that there is an interaction between the genetic vulnerability and environmental factors in the aetiology and course of the disorder. Environmental factors have been neglected for a long time because bipolar disorder was considered an endogenous disease. Thus, very recently, studies have shown that early trauma could contribute to the occurrence or the severity of the disease. During a lifetime, many environmental components (stressful life events, changes in daily schedules, drug abuse, some medications) could also trigger the onset of the pathology or the occurrence of new episodes. It is very important to recognize these factors in order to develop preventive strategies for limiting their impact. PMID- 15895951 TI - [Clinical picture and diagnosis of bipolar disorder]. AB - Since more than one century, the illness considerably evolved. Between the extremes of full-blown manic-depressive disorder and the strictly defined unipolar depression, patients exhibit particular affective conditions characterised by complex or soft symptoms. So the actual classification encompasses 6 clinical forms with sometimes, sub-forms. In this context, an awareness of the different clinical presentations is an important issue in the daily practice, especially hypomania, cyclothymia, mixed states and affective episode with delusion. These conditions are often misdiagnosed and the bipolar trouble is under-diagnosed. Although, the lifetime prevalence of the classical manic-depressive is about 1%, current evidences indicate that the bipolar spectrum account for 5-8% in the general population. So, the public health significance of a rapid and proper diagnosis is quite essential because the treatment needs to be applied in the clinical care. PMID- 15895952 TI - [Bipolar disorders and psychiatric comorbidity]. AB - Comorbidity is very frequent in bipolar disorders. The most frequently psychiatric disorders associated with bipolar disorders are substance use disorders (particularly alcohol abuse) and anxiety disorders. Obsessive compulsive disorder, impulse control disorders, personality disorders and eating disorders co-occur also with bipolar disorder. Comorbidity complicates the diagnosis and the treatment of bipolar disorder and worsen the prognosis. For every bipolar patient, it is necessary to search for a comorbid psychiatric disorder, and vice-versa. There is a noticeable paucity of controlled studies to guide selection of the most effective treatment approches for these patients. The expert consensus guidelines about treatment of bipolar disorder propose divalproex as first-line treatment when bipolar disorder co-occurres with substance abuse, panic disorder or posttraumatic stress disorder. PMID- 15895953 TI - [Clinical masks of bipolar disorders]. AB - Bipolar disorders usually present as alternating episodes of melancholy and mania. However, other clinical situations appear in a masked form before being spotted. The principal one is hypomania that declares itself as an attenuated form of a manic episode. Its principal signs are an increase in energy, hyperactivity and disinhibition. In other cases, bipolar disorders express themselves as impulsive behaviours. More rarely, the mask of a bipolar illness is translated into behaviours such as intermittent alcohol abuse. The consummation of alcohol is sudden and paroxysmal. It is a type of dipsomania. Finally, bipolar disorders can express themselves as difficulty in impulse control. The two most frequent examples are impulse buying and pathological gambling. PMID- 15895954 TI - [Management of bipolar disorders]. AB - Management of the bipolar disorders is well codified and rests on the prescription of mood stabilisers and psychotherapeutic measures. Mood stabilisers have considerably improved the prognosis of bipolar disorder, in limiting the number of recurrences. Lithium is still the leader of mood stabilisers. It has a preventative and curative action that has effect on the two poles of the illness. Carbamazepine, valpromide and its derivatives, and olanzapine are indicated in the case of resistance or contra-indication to lithium therapy and are more easily handled in the curative treatment of the manic state. Other therapeutic measures have been proposed in order to optimise the medical treatment and to act in anticipation of the precipitating elements. Psycho educational measures, the roles of which have been undervalued for a long time, are today the best evaluated psychological treatments. An elevated level of proof exists for these treatments. Their value is stressed in most of the reference texts. The benefits of this complementary approach exists on different levels: early recognition of symptoms that announce a recurrence, improvement in compliance, acceptance of the illness, better management of the social life, both professional and emotional, control of precipitating factors... Recently published studies report a reduction in the number of recurrences and relapses, a reduction in the duration of hospitalisation, a better balance in the family life and an improvement in compliance. PMID- 15895955 TI - [Which signs in a depressed patient could indicate a manic transformation?]. AB - Which signs in a depressed patient could indicate a manic transformation? All depressive states can give way to a state of manic excitation. The transformation can be spontaneous or favoured by antidepressants. The principal indices of risk of mood transformation are a personal or family history of mood disorders, thymic lability, marked psychomotor inhibition, the existence of overeating and hypersomnia, early age of onset, acute onset and resolution of episodes. It is important in the presence of these elements to re-evaluate the indication for an antidepressant and to consider the prescription of a mood stabiliser as monotherapy or in association with an antidepressant. PMID- 15895956 TI - [Bipolar disorder in MEDLINE]. PMID- 15895957 TI - [Do you suffer from bipolar disorder?]. PMID- 15895958 TI - [Continuing medical education: a growth under control?]. PMID- 15895959 TI - [Nutritional problems in the child and the adult]. PMID- 15895960 TI - [The search for risk factors and the therapeutic consequences. Drug interactions]. PMID- 15895961 TI - [Recent neurological episodes]. PMID- 15895962 TI - [Principles of a program of quality assurance and evaluation of professional practices]. PMID- 15895963 TI - [Treating victims of sexual violence]. PMID- 15895964 TI - [Representations of uterine life on Graeco-Roman magical gems. See the invisible]. PMID- 15895965 TI - [Pharmaceutical hydrophilic gels]. AB - From the pharmacopoeial standpoint, gels, together with ointments, creams, pastes, cataplasmata, and medicated plasters, rank among the group of topical semisolid preparations applied to the skin. They are bicoherent systems composed of the internal phase made of a polymer producing a coherent three-dimensional net-like structure, which fixes the liquid vehicle as the external phase. Intermolecular forces bind the molecules of the solvent to a polymeric net, thus decreasing the mobility of these molecules and producing a structured system with increased viscosity. The physical and chemical bonds binding the particles of the internal phase provide a relatively stable structure, which can originate by swelling of solid polymers, or by decreasing the solubility of the polymer in a solution. An important group of gels used in pharmacy are hydrophylic gels, or hydrogels, usually made of hydrophyilc polymers, which under certain conditions and polymer concentration, jellify. Attention of pharmaceutical research now concentrates primarily on hydrophilic gels, as this dosage form seems to be prospective for the development of modern drugs based on systems with prolonged and controlled release of active ingredients. PMID- 15895966 TI - [Adsorption of substances on active charcoal--part 3: stobadin acylderivatives]. AB - The paper studies the adsorption of nine stobadin acylderivatives on active charcoal. Freundlich model of adsorption was employed to evaluate the course of adsorption in dependence on the concentration of substances. At the same time, the dependence of adsorption on lipophilicity and structural parameters of substances was evaluated. The dependence of the amount of the adsorbed substance on time served for the calculation of the rate constant of the 1st order of adsorption process. These values correlate with the parameters characterizing the size of the molecule. PMID- 15895967 TI - [Dispensation during pharmaceutical emergency service]. AB - The paper deals with the analysis of 24 694 medical prescriptions dispensed during pharmaceutical stand-by duty service within 12 months. 8072 medical prescriptions (32.7 %) were issued by first-aid medical service. 72.2 % of requirements for pharmaceutical stand-by duty service took place between 19.00 to 22.00. Patients under 20 requiring pharmaceutical stand-by duty service represented 37.8 %. One medical prescription issued by medical first-aid service included 1.8 packages of pharmaceutical preparations. Medical prescriptions issued by medical first-aid service to adults included altogether 529 different sorts of medicinal preparations, those issued to children, 269 sorts. PMID- 15895968 TI - [Dispensing care at the dispensing counter in the conditions of a Czech pharmacy]. AB - Evaluation of 108 cases when the pharmacist found a need to intervene into prescription verified the use of internationally employed system of classification of pharmaceutical interventions in the conditions of the Czech pharmacy of the basic type. The so-called dispensing care at the counter was provided by five participating pharmacists (out of seven working in the pharmacy) for a period of six months. The most frequently identified problems included: inappropriate use of medicaments--overdose, use at unsuitable time (28.7 %), a need to inform the patient (17.6 %), potentially ineffective therapy (12.9 %), drug interactions (7.4 %), suspected and potential undesirable effects of prescribed medicaments (7.4 %), and insufficient compliance (6.5 %). Solution of problems with medicaments was most frequently: a change in dosage (25.9 %), recommendation to consult the physician (24.1 %), and professional counselling on pharmacotherapy (23.1 %). The pharmacotherapeutic group identified as the one most frequently requiring a telephone consultation of the pharmacist and the physician was antibiotics, including antibacterial chemotherapeutic agents. PMID- 15895969 TI - [Preformulation studies of potential drug XIX M: partition coefficient]. AB - The factors which markedly influence availability of the drug from the dosage form include also auxiliary substances, which are an inevitable component in the formulation of the drug. In the selection of auxiliary substances for a newly formulated drug, it is necessary to know that the drug is never a simple mixture of mutually independent ingredients, but a dynamic system in which various physical and chemical processes take place. The present paper aims to study the effect of auxiliary substances from the group of humectants with graded concentrations and the effect of the preservative on the partition coefficient of potential drug XIX M. Partition coefficient (P') was estimated in the system n octanol/aqueous solution with graded concentrations of polyols. In these estimations, n-octanol simulated the horny layer, and the aqueous solution the base of the topical preparation. The auxiliary substances employed were polyols - glycerol, propylene glycol, and sorbitol in 5, 10, 15, and 20% concentrations and an antimicrobially effective solution of Ajatin (Solution benzododecinii bromati) in two concentrations of 0.01 and 0.1 wt %. It follows from the obtained results that the values of partition coefficient of potential drug XIX M are greatly influenced by auxiliary substances. The value of this parameter, and therefore also biological availability, depend not only on the sort of the polyol used and its concentration, but also on the concentration of the preservative employed, in this case Ajatin. PMID- 15895970 TI - [Arbutin, salicin: the possibilities of their biotechnological production]. AB - The paper aimed to transform exogenous precursors with in vitro cultures of Datura meteloides, Coronilla varia, Leuzea carthamoides and Schisandra chinensis. These cultures were added the precursors of arbutin and salicin (phenylalanine, cinnamic, p-coumaric, p-anisoic, o-coumaric, salicylic acids, salicylaldehyde, helicin), not yet tested by the present authors. The culture of Schisandra chinensis was also added, besides the above-mentioned precursors, hydroquinone, because this culture had not been employed for biotransformation purposes yet. The precursors tested were used in a concentration of 100 mg x l(-1) and the period of their action was 6; 12; 24; 48, and 168 hours. Positive results (both TLC and HPLC) in arbutin production were obtained in the culture of Schisandra chinensis after an addition of hydroquinone. The largest amount of arbutin in callus cultures was measured after a week's cultivation with hydroquinone (5.08 %). In this experimental variant, arbutin was released also to the culture medium. Our results revealed salicylaldehyde to be the optimal precursor of salicin. It was transformed by the culture of Datura meteloides after 6; 24, and 168 hours and by the culture of Coronilla varia after 6 hours. In comparison with arbutin, its amount was smaller. PMID- 15895971 TI - [Use of dinitrophenols in ion-pair extraction spectrophotometry of cationic tensides]. AB - The application of 2,4-dinitrophenol and 2,6-dinitrophenol as reagents for a ion pair extraction spectrophotometry was tested. The ion pair extraction spectrophotometry experiments were carried out using a simplified procedure (test tube method). Four cationic tensides were used as model analytes. After the optimal conditions of the assay were being determined, the influence of the concentration excess of the reagents on the absorbance of the chloroform extract was investigated. Calibration curves were constructed; an estimation of limits of quantitation was performed and apparent molar absorption coefficients were determined. The obtained results show that both investigated dinitrophenols are suitable for the studied purpose. PMID- 15895972 TI - [Use of capillary isotachophoresis for the determination of pheniramine in granulated powders]. AB - The method of capillary isotachophoresis was used to assay pheniramine in granulated powders. Several electrolyte systems of different composition and different pH were tested. Two electrolyte systems were selected for the validation of the method and pheniramine determination in real samples. Precision, accuracy, linearity, robustness, and selectivity of the ITP method for both electrolyte systems were evaluated. The pre-treatment of samples prior to analysis consisted in dissolving pheniramine-containing granulated powder in demineralized water and subsequent diluting with demineralized water to required concentration. PMID- 15895973 TI - [Pharmacoeconomic profile of antibiotics used in the treatment of lower respiratory tract infections in geriatric patients]. AB - The paper aims to evaluate the pharmacoeconomic profile of antibiotics (ATB) used in the treatment of lower respiratory tract (LRT) infections and thus contribute to rationalization of therapeutic procedures. Of 2870 patients hospitalized at the Geriatric Clinic of the Medical Faculty of Comenius University from 1 January 1999 to 31 December 2001, 189 patients with acute infections of the LRT were included in the retrospective study. For pharmacoeconomic evaluation, cost effectiveness analysis was employed, the principal parameter of which, cost effectiveness coefficient, was the ratio of the price of ATB treatment in Slovak crowns (SK) and the criterion of effectiveness (E), the number of asymptomatic days in a month. The authors separately evaluated ATBs administered perorally (p.o.); intravenously (i.v.), and sequentially, and they also compared i.v. and the corresponding sequentially administered ATBs. Statistical comparison revealed significant differences in the prices and cost effectiveness coefficients of individual alternatives of ATB treatment. Employed ATBs did not significantly differ in the criteria of effectiveness. According to the cost effectiveness coefficient (SK/asymptomatic day), fluoroquinolones were less expensive (median price/E: pefloxacin: p.o. 19.3; i.v. 29.1; sequentially administered 26.0, and ciprofloxacin: p.o., 14.7, i.v., 54.1, sequentially administered, 31.7). Sequential administration of ATBs (ampicillin-sulbactam, cefuroxime, amoxicillin klavulanate, ciprofloxacin) was significantly cheaper in comparison with i.v. administration. With therapeutic equivalence, the total pharmacoeconomic profile of ATB treatment depended on the price parameter. In the selection of ATB it is also necessary to consider the price of the drug. PMID- 15895974 TI - [Effect of polymer concentration on rheological properties of Carbopol 980 hydrogels containing trimecainium chloride]. AB - Rheological properties exert influence on the manufacturing processes, stability and usability of a dosage form as well as the biological availability of an active ingredient. The present paper examines the effect of Carbopol 980 concentration on rheological properties of hydrogels with and without trimecainium chloride for a period of 2, 7, and 14 days after their preparation. On the basis of an evaluation of the changes in structural viscosity in dependence on the period of measurement, it has been concluded that for formulation of trimecainium chloride to a dermal dosage form, Carbopol 980 concentrations of 0.5 and 1.2 % are suitable. PMID- 15895975 TI - Why enzymes are proficient catalysts: beyond the Pauling paradigm. AB - Pauling proposed that "enzymes are molecules that are complementary in structure to the activated complexes of the reactions that they catalyze, ..., [rather than] entering into reactions". This paradigm has dominated thinking in the field. While complementarity of the type proposed by Pauling can account for acceleration up to 11 orders of magnitude, most enzymes exceed that proficiency. Enzymes with proficiencies ((k(cat)/K(M))/k(uncat)) > 10(11) M(-1) achieve over 15 kcal/mol of "transition state binding" not merely by a concatenation of noncovalent effects but by covalent bond formation between enzyme or cofactor and transition state, involving a change in mechanism from that in aqueous solution. Enzymes enter into reactions with substrates and do not merely complement the transition states of the uncatalyzed reactions. PMID- 15895976 TI - Halogen bonding based recognition processes: a world parallel to hydrogen bonding. AB - Halogen bonding is the noncovalent interaction between halogen atoms (Lewis acids) and neutral or anionic Lewis bases. The main features of the interaction are given, and the close similarity with the hydrogen bonding will become apparent. Some heuristic principles are presented to develop a rational crystal engineering based on halogen bonding. The focus is on halogen-bonded supramolecular architectures given by halocarbons. The potential of the interaction is shown by useful applications in the field of synthetic chemistry, material science, and bioorganic chemistry. PMID- 15895977 TI - Reflection on medium effects on photochemical reactivity. AB - In reexamining medium effects on photochemical reactions, we have emphasized those on unequilibrated excited species such as the Franck-Condon species. Despite recent advances in femtochemistry, such a discussion in molecular photochemistry is uncommon, and the problem remains challenging on account of the extremely short-lived excited species. However, in such cases, a small perturbation resulting from, for example, weak guest-host interactions could turn into a determining factor in dictating the course of a photochemical channel of deactivation. Examples of medium-directed diabatic processes have been examined with this idea in mind. A modified view on rhodopsin photoisomerization is presented along with the consideration that confinement does not necessarily lead to inhibition of reactions of the trapped substrate. PMID- 15895978 TI - Conformationally constrained PNA analogues: structural evolution toward DNA/RNA binding selectivity. AB - Since its discovery 12 years ago, aminoethylglycyl peptide nucleic acid (aeg-PNA) has emerged as one of the successful DNA mimics for potential therapeutic and diagnostic applications. An important requisite for in vivo applications that has received inadequate attention is engineering PNA analogues for able discrimination between DNA and RNA as binding targets. Our approach toward this aim is based on structural preorganization of the backbone to hybridization competent conformations to impart binding selectivity. This strategy has allowed us to design locked PNAs to achieve specific hybridization with DNA or RNA with aims to increase the binding strength without losing the binding specificity. This Account presents results of our rationale in design of different conformationally constrained PNA analogues, their synthesis, and evaluation of hybridization specificities. PMID- 15895979 TI - The virtue of palladium-catalyzed domino reactions - diverse oligocyclizations of acyclic 2-bromoenynes and 2-bromoenediynes. AB - The domino-type combination of consecutive palladium-catalyzed cross-coupling reactions with subsequent pericyclic transformations offers a very fast access to various oligocyclic skeletons. This Account highlights all-intramolecular, intra intermolecular, and all-intermolecular organopalladation cascades leading to 1,3,5-hexatrienes, which undergo consecutive 6pi-electrocyclizations with closure of three, two, or just one ring, respectively. Sequences of cross-coupling reactions and Diels-Alder additions are also discussed, as well as tricyclizations of 2-bromoenediynes giving rise to bisannelated benzene and fulvene derivatives. PMID- 15895980 TI - Synthetic strategies of marine polycyclic ethers via intramolecular allylations: linear and convergent approaches. AB - Strategies for the synthesis of polycyclic ethers based on intramolecular allylations are overviewed. The intramolecular condensation of allylic stannanes and aldehydes is a powerful tool for the synthesis of oxepane derivatives. The reaction is successfully applied to the iterative total synthesis of hemibrevetoxin B (2). Further, the intramolecular allylation of alpha-acetoxy ethers provides an efficient method for the convergent synthesis of polycyclic ethers. The usefulness of the latter strategy is demonstrated in the convergent total synthesis of gambierol (4). PMID- 15895981 TI - Fifteen years of Raman spectroscopy of engineered heme containing peroxidases: what have we learned? AB - Spectroscopic techniques have been fundamental to the comprehension of peroxidase function under physiological conditions. This Account examines the contribution to our understanding of heme peroxidases provided by electronic and resonance Raman spectroscopies in conjunction with site-directed mutagenesis. The results obtained over 15 years with several heme peroxidases and selected mutants have provided important insights into the influence exerted by the protein in the vicinity of the active site via key amino acids on the functionality and stability of the enzymes. Moreover, resonance Raman spectroscopy has revealed that a common feature of heme peroxidases is the presence of an extensive network of H-bonds coupling the distal and proximal sides, which has a profound influence on the heme ligation, affecting both the fifth and the sixth coordination sites. PMID- 15895982 TI - Design of a redox-linked active metal site: manganese bound to bacterial reaction centers at a site resembling that of photosystem II. AB - Metals bound to proteins perform a number of crucial biological reactions, including the oxidation of water by a manganese cluster in photosystem II. Although evolutionarily related to photosystem II, bacterial reaction centers lack both a strong oxidant and a manganese cluster for mediating the multielectron and proton transfer needed for water oxidation. In this study, carboxylate residues were introduced by mutagenesis into highly oxidizing reaction centers at a site homologous to the manganese-binding site of photosystem II. In the presence of manganese, light-minus-dark difference optical spectra of reaction centers from the mutants showed a lack of the oxidized bacteriochlorophyll dimer, while the reduced primary quinone was still present, demonstrating that manganese was serving as a secondary electron donor. On the basis of these steady-state optical measurements, the mutant with the highest affinity site had a dissociation constant of approximately 1 microM. For the highest-affinity mutant, a first-order rate with a lifetime of 12 ms was observed for the reduction of the oxidized bacteriochlorophyll dimer by the bound manganese upon exposure to light. The dependence of the amplitude of this component on manganese concentration yielded a dissociation constant of approximately 1 muM, similar to that observed in the steady-state measurements. The three-dimensional structure determined by X-ray diffraction of the mutant with the high-affinity site showed that the binding site contains a single bound manganese ion, three carboxylate groups (including two groups introduced by mutagenesis), a histidine residue, and a bound water molecule. These reaction centers illustrate the successful design of a redox active metal center in a protein complex. PMID- 15895983 TI - Human glucocorticoid receptor alpha transcript splice variants with exon 2 deletions: evidence for tissue- and cell type-specific functions. AB - Alternative splicing of exon 9 in human glucocorticoid receptor (hGR) transcripts yields two native hGR transcripts and proteins, hGRalpha and hGRbeta. We have now identified four novel hGRalpha transcripts that have various deletions of exon 2 sequences. Among these hGRalpha splice variants, three of them, 1A1/E2dist hGRalpha, 1A2/E2prox hGRalpha, and 1A3/E3 hGRalpha, arise from the hGR 1A promoter, while 1B/E3 hGRalpha comes from the hGR 1B promoter. When fused to Flag and enhanced green fluorescent protein (EGFP) tags at the carboxy terminus, all transcript variants can be correctly translated in vitro and in vivo. The Flag tagged hGRalpha protein variants can functionally bind to a glucocorticoid response element (GRE) and can mediate hormonal stimulation of a pGRE-luciferase reporter gene. Compared to the "classical", native hGRalpha, these four variants exhibit a cell type-specific activation of a reporter gene, and this is influenced by the hGRalpha 3' untranslated region in the hGR transcript. When equal amounts of the cDNAs for these GRalpha variant proteins are transfected into cells, they can exhibit lower or higher transcriptional activation compared to the classical GR. Furthermore, the EGFP-tagged proteins are nuclear localized, even in the absence of hormone. Using quantitative reverse transcription PCR, we found that these transcripts exist at a low level in CEM-C7 cells and IM-9 cells, although the concentrations of the 1A3/E3 hGRalpha and 1B/E3 hGRalpha transcripts are higher than for hGRbeta transcripts, while 1A1/E2dist hGRalphaand 1A2/E2prox hGRalpha transcript levels are comparable to the 1A1 hGRalpha and 1A2 hGRalpha (without the exon 2 deletions) transcript levels, respectively. Because these novel hGR, N-terminal deleted, protein variants have altered biological activity, their expression could potentially affect the hormone sensitivity or resistance in leukemia and be useful in diagnosing hormone-sensitive or -resistant disease. PMID- 15895984 TI - FTIR studies of internal water molecules in the Schiff base region of bacteriorhodopsin. AB - In a light-driven proton pump protein, bacteriorhodopsin (BR), three water molecules participate in a pentagonal cluster that stabilizes an electric quadrupole buried inside the protein. Previously, low-temperature Fourier transform infrared (FTIR) difference spectra between BR and the K photointermediate in D(2)O revealed six O-D stretches of water in BR at 2690, 2636, 2599, 2323, 2292, and 2171 cm(-)(1), while five water bands were observed at 2684, 2675, 2662, 2359, and 2265 cm(-)(1) for the K intermediate. The frequencies are widely distributed over the possible range of stretching vibrations of water, and water molecules at <2400 cm(-)(1) were suggested to hydrate negative charges because of their extremely strong hydrogen bonds. In this paper, we aimed to reveal the origin of these water bands in the K minus BR spectra by use of various mutant proteins. The water bands were not affected by the mutations at the cytoplasmic side, such as T46V, D96N, and D115N, implying that the water molecules in the cytoplasmic domain do not change their hydrogen bonds in the BR to K transition. In contrast, significant modifications of the water bands were observed for the mutations in the Schiff base region and at the extracellular side, such as R82Q, D85N, T89A, Y185F, D212N, R82Q/D212N, and E204Q. From these results, we concluded that the six O-D stretches of BR originate from three water molecules, water401, -402, and -406, involved in the pentagonal cluster. Two stretching modes of each water molecule are highly separate (300-470 cm(-)(1) for O-D stretches and 500-770 cm(-)(1) for O-H stretches), which is consistent with the previous QM/MM calculation. The small amplitudes of vibrational coupling are presumably due to strong association of the waters to negative charges of Asp85 and Asp212. Among various mutant proteins, only D85N and D212N lack strongly hydrogen-bonded water molecules (<2400 cm(-)(1)) and proton pumpimg activity. We thus infer that the presence of a strong hydrogen bond of water is a prerequisite for proton pumping in BR. Internal water molecules in such a specific environment are discussed in terms of functional importance for rhodopsins. PMID- 15895985 TI - Structural difference between group I and group II cobra cardiotoxins: X-ray, NMR, and CD analysis of the effect of cis-proline conformation on three-fingered toxins. AB - Natural homologues of cobra cardiotoxins (CTXs) were classified into two structural subclasses of group I and II based on the amino acid sequence and circular dichroism analysis, but the exact differences in their three-dimensional structures and biological significance remain elusive. We show by circular dichroism, NMR spectroscopic, and X-ray crystallographic analyses of a newly purified group I CTX A6 from eastern Taiwan cobra (Naja atra) venoms that its loop I conformation adopts a type VIa turn with a cis peptide bond located between two proline residues of PPxY. A similar "banana-twisted" conformation can be observed in other group I CTXs and also in cyclolinopeptide A and its analogues. By binding to the membrane environment, group I CTX undergoes a conformational change to adopt a more extended hydrophobic domain with beta-sheet twisting closer to the one adopted by group II CTX. This result resolves a discrepancy in the CTX structural difference reported previously between solution as well as crystal state and shows that, in addition to the hydrophobicity, the exact loop I conformation also plays an important role in CTX-membrane interaction. Potential protein targets of group I CTXs after cell internalization are also discussed on the basis of the determined loop I conformation. PMID- 15895986 TI - Comparative investigation of the LOV1 and LOV2 domains in Adiantum phytochrome3. AB - Phototropin (phot) is a blue-light photoreceptor for phototropic responses, relocation of chloroplasts, and stomata opening in plants. Phototropin has two chromophore-binding domains named LOV1 and LOV2 in its N-terminal half, each of which binds a flavin mononucleotide (FMN) noncovalently. The C-terminal half is a Ser/Thr kinase. A transgenic study of Arabidopsis suggested that only LOV2 domain is necessary for the kinase activity, whereas X-ray crystallographic structures of LOV1 and LOV2 domains are almost identical. These facts imply that the detailed structures and/or structural changes are different between LOV1 and LOV2 domains. In this study, we compared light-induced structural changes of the LOV1 and LOV2 domains of a phototropin, Adiantum phytochrome3 (phy3), by means of UV visible and Fourier transform infrared (FTIR) spectroscopy. Photochemical properties of an adduct formation between FMN and a cysteine are essentially similar between phy3-LOV1 and phy3-LOV2. On the other hand, the S-H group of the reactive cysteine forms a hydrogen bond in phy3-LOV1, which is strengthened at low temperatures. This is possibly correlated with the fact that no adduct formation takes place for phy3-LOV1 at 77 K as revealed by the UV-visible absorption spectra. The most prominent difference was seen in the amide-I vibration that monitors the secondary structure of peptide backbone. Protein structural changes in phy3-LOV2 involve the regions of loops, alpha-helices, and beta-sheets, which differ significantly among various temperatures. Extended protein structural changes are probably correlated with the signal transduction activity of LOV2. In contrast, protein structural changes were very small in phy3 LOV1, and they were almost temperature independent. The photocycle of phy3-LOV1 takes 3.1 h, being more than 100 times longer than that of phy3-LOV2. These facts suggest that Adiantum phy3-LOV1 does not work for light sensing, being consistent with the previous transgenic study of Arabidopsis. It is likely that plants utilize a unique protein architecture (LOV domain) for different functions by regulating their protein structural changes. PMID- 15895987 TI - Robustness of downhill folding: guidelines for the analysis of equilibrium folding experiments on small proteins. AB - Previously, we identified the protein BBL as a downhill folder. This conclusion was based on the statistical mechanical analysis of equilibrium experiments performed in two variants of BBL, one with a fluorescent label at the N-terminus, and another one labeled at both ends. A recent report has claimed that our results are an artifact of label-induced aggregation and that BBL with no fluorescent labels and a longer N-terminal tail folds in a two-state fashion. Here, we show that singly and doubly labeled BBL do not aggregate, unfold reversibly, and have the same thermodynamic properties when studied under appropriate experimental conditions (e.g., our original conditions (1)). With an elementary analysis of the available data on the nonlabeled BBL (2), we also show that this slightly more stable BBL variant is not a two-state folder. We discuss the problems that led to its previous misclassification and how they can be avoided. Finally, we investigate the equilibrium unfolding of the singly labeled BBL with both ends protected by acetylation and amidation. This variant has the same thermodynamic stability of the nonlabeled BBL and displays all the equilibrium signatures of downhill folding. From all these observations, we conclude that fluorescent labels do not perturb the thermodynamic properties of BBL, which consistently folds downhill regardless of its stability and specific protein tails. The work on BBL illustrates the shortcomings of applying conventional procedures intended to distinguish between two-state and three-state folding models to small fast-folding proteins. PMID- 15895988 TI - 2.3 A crystal structure of tetanus neurotoxin light chain. AB - TeNT is the causative agent of the neuroparalytic disease tetanus. A key component of TeNT is its light chain, a Zn(2+) endopeptidase that targets SNAREs. Recent structural studies of closely related BoNT endopeptidases indicate that substrate-binding exosites remote from a conserved active site are the primary determinants of substrate specificity. Here we report the 2.3 A X-ray crystal structure of TeNT-LC, determined by combined molecular replacement and MAD phasing. As expected, the overall structure of TeNT-LC is similar to the other known CNT light chain structures, including a conserved thermolysin-like core inserted between structurally distinct amino- and carboxy-terminal regions. Differences between TeNT-LC and the other CNT light chains are mainly limited to surface features such as unique electrostatic potential profiles. An analysis of surface residue conservation reveals a pattern of relatively high variability matching the path of substrate binding around BoNT/A, possibly serving to accommodate the variations in different SNARE targets of the CNT group. PMID- 15895989 TI - Structure of B-DNA with cations tethered in the major groove. AB - Here, we describe the 1.6-A X-ray structure of the DDD (Dickerson-Drew dodecamer), which has been covalently modified by the tethering of four cationic charges. This modified version of the DDD, called here the DDD(4+), is composed of [d(CGCGAAXXCGCG)](2), where X is effectively a thymine residue linked at the 5 position to an n-propyl-amine. The structure was determined from crystals soaked with thallium(I), which has been broadly used as a mimic of K(+) in X-ray diffraction experiments aimed at determining positions of cations adjacent to nucleic acids. Three of the tethered cations are directed radially out from the DNA. The radially directed tethered cations do not appear to induce structural changes or to displace counterions. One of the tethered cations is directed in the 3' direction, toward a phosphate group near one end of the duplex. This tethered cation appears to interact electrostatically with the DNA. This interaction is accompanied by changes in helical parameters rise, roll, and twist and by a displacement of the backbone relative to a control oligonucleotide. In addition, these interactions appear to be associated with displacement of counterions from the major groove of the DNA. PMID- 15895990 TI - Relationship between folding and function in a sequence-specific miniature DNA binding protein. AB - Previously, we have described a miniature protein-based approach to the design of molecules that bind DNA or protein surfaces with high affinity and specificity. In this approach, the small, well-folded protein avian pancreatic polypeptide acts as a scaffold to present and stabilize an alpha-helical or PPII-helical recognition epitope. The first miniature protein designed in this way, a molecule called p007, presents the alpha-helical recognition epitope found on the bZIP protein GCN4 and binds DNA with nanomolar affinity and exceptional specificity. In this work we use alanine-scanning mutagenesis to explore the contributions of 29 p007 residues to DNA affinity, specificity, and secondary structure. Virtually every residue within the p007 alpha-helix, and most residues within the p007 PPII helix, contribute to both DNA affinity and specificity. These residues include those introduced to make specific and nonspecific DNA contacts, as well as those that complete the miniature protein core. Moreover, there exists a direct correlation between the affinity of a p007 variant for specific DNA and the ability of that variant to select for specific DNA over nonspecific DNA. Although we observe no correlation between alpha-helicity and affinity, we observe a limited correlation between alpha-helicity and sequence specificity that emphasizes the role of coupled binding/folding in the function of p007. Our results imply that formation of a highly evolved set of protein.DNA contacts in the context of a well-packed hydrophobic core, and not the extent of intrinsic alpha-helical structure, is the primary determinant of p007 function. PMID- 15895991 TI - Raft partitioning and dynamic behavior of human placental alkaline phosphatase in giant unilamellar vesicles. AB - Much attention has recently been drawn to the hypothesis that cellular membranes organize in functionalized platforms called rafts, enriched in sphingolipids and cholesterol. The notion that glycosylphosphatidylinositol (GPI)-anchored proteins are strongly associated with rafts is based on their insolubility in nonionic detergents. However, detergent-based methodologies for identifying raft association are indirect and potentially prone to artifacts. On the other hand, rafts have proven to be difficult to visualize and investigate in living cells. A number of studies have demonstrated that model membranes provide a valuable tool for elucidating some of the raft properties. Here, we present a model membrane system based on domain-forming giant unilamellar vesicles (GUVs), in which the GPI-anchored protein, human placental alkaline phosphatase (PLAP), has been functionally reconstituted. Raft morphology, protein raft partitioning, and dynamic behavior have been characterized by fluorescence confocal microscopy and fluorescence correlation spectroscopy (FCS). Approximately 20-30% of PLAP associate with sphingomyelin-enriched domains. The affinity of PLAP for the liquid-ordered (l(o)) phase is compared to that of a nonraft protein, bacteriorhodopsin. Next, detergent extraction was carried out on PLAP-containing GUVs as a function of temperature, to relate the lipid and protein organization in distinct phases of the GUVs to the composition of detergent resistant membranes (DRMs). Finally, antibody-mediated cross-linking of PLAP induces a shift of its partition coefficient in favor of the l(o) phase. PMID- 15895992 TI - Structural characterization of the molten globule state of apomyoglobin by limited proteolysis and HPLC-mass spectrometry. AB - A method to characterize the structural conformation of an acidic molten globule apomyoglobin (apoMb) at pH 4.2 was developed using limited proteolysis and HPLC mass spectrometry (HPLC-MS). Endoproteinase Glu-C, which has a double maximum activity at pH 4.0 and pH 7.8 toward glutamic acid (Glu), was used as a proteolytic enzyme. Using this method enabled us to compare the proteolytic cleavages of native apoMb (at pH 8.0) and molten globule (at pH 4.2) directly. Only the first cleavage event in each molecule was considered as reflecting original structural information since the original structure of the protein can be altered after the fist cleavage. Structural changes of apoMb in various pH conditions were studied here to elucidate the local helicity of molten globule apoMb. Among 13 Glu sites, only Glu83 and Glu85 in the F-helix were cleaved at pH 8.0, which confirms that only helix F is frayed upon removal of heme group. At acidic molten globule state, rapid cleavages at Glu38, Glu52, Glu54, Glu85, and Glu148 were detected, while the remaining eight sites were protected. Glu6 and Glu18 in the A-helix, and Glu105 in the G-helix were protected due to the helicity of the secondary structures. The cleavage at Glu38 and the protection at Glu41 in the C-helix indicate that the first half of the C-helix is frayed and the second half of the C-helix is structured. Cleavage at both Glu52 and Glu54 in the D-helix proves that the D-helix is disordered. The N-terminal end of the E helix at Glu59 was protected, and the beginning of the F-helix was protected by aid of the pH-induced C-cap of the E-helix. The cleavage at Glu148 in H suggests that the C-terminal end of the H-helix is disordered. The A-helix and the first half of the B-helix were highly stable. PMID- 15895993 TI - Solution structure of the K50 class homeodomain PITX2 bound to DNA and implications for mutations that cause Rieger syndrome. AB - We have determined the solution structure of a complex containing the K50 class homeodomain Pituitary homeobox protein 2 (PITX2) bound to its consensus DNA site (TAATCC). Previous studies have suggested that residue 50 is an important determinant of differential DNA-binding specificity among homeodomains. Although structures of several homeodomain-DNA complexes have been determined, this is the first structure of a native K50 class homeodomain. The only K50 homeodomain structure determined previously is an X-ray crystal structure of an altered specificity mutant, Engrailed Q50K (EnQ50K). Analysis of the NMR structure of the PITX2 homeodomain indicates that the lysine at position 50 makes contacts with two guanines on the antisense strand of the DNA, adjacent to the TAAT core DNA sequence, consistent with the structure of EnQ50K. Our evidence suggests that this side chain may make fluctuating interactions with the DNA, which is complementary to the crystal data for EnQ50K. There are differences in the tertiary structure between the native K50 structure and that of EnQ50K, which may explain differences in affinity and specificity between these proteins. Mutations in the human PITX2 gene are responsible for Rieger syndrome, an autosomal dominant disorder. Analysis of the residues mutated in Rieger syndrome indicates that many of these residues are involved in DNA binding, while others are involved in formation of the hydrophobic core of the protein. Overall, the role of K50 in homeodomain recognition is further clarified, and the results indicate that native K50 homeodomains may exhibit differences from altered specificity mutants. PMID- 15895994 TI - Kinetics of the inhibition of bovine liver dihydrofolate reductase by tea catechins: origin of slow-binding inhibition and pH studies. AB - Dihydrofolate reductase (DHFR) is the subject of intensive investigation since it appears to be the primary target enzyme for "antifolate" drugs, such as methotrexate and trimethoprim. Fluorescence quenching and stopped-flow fluorimetry show that the ester bond-containing tea polyphenols (-) epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG) are potent and specific inhibitors of DHFR with inhibition constants (K(I)) of 120 and 82 nM, respectively. Both tea compounds showed the characteristics of slow-binding inhibitors of bovine liver DHFR. In this work, we have determined a complete kinetic scheme to explain the slow-binding inhibition and the pH effects observed during the inhibition of bovine liver DHFR by these tea polyphenols. Experimental data, based on fluorimetric titrations, and transient phase and steady-state kinetic studies confirm that EGCG and ECG are competitive inhibitors with respect to 7,8-dihydrofolate, which bind preferentially to the free form of the enzyme. The origin of their slow-binding inhibition is proposed to be the formation of a slow dissociation ternary complex by the reaction of NADPH with the enzyme inhibitor complex. The pH controls both the ionization of critical catalytic residues of the enzyme and the protonation state of the inhibitors. At acidic pH, EGCG and ECG are mainly present as protonated species, whereas near neutrality, they evolve toward deprotonated species due to ionization of the ester-bonded gallate moiety (pK = 7.8). Although DHFR exhibits different affinities for the protonated and deprotonated forms of EGCG and ECG, it appears that the ionization state of Glu-30 in DHFR is critical for its inhibition. The physiological implications of these pH dependencies are also discussed. PMID- 15895995 TI - Engineering intracellular CMP-sialic acid metabolism into insect cells and methods to enhance its generation. AB - Previous studies have reported that insect cell lines lack the capacity to generate endogenously the nucleotide sugar, CMP-Neu5Ac, required for sialylation of glycoconjugates. In this study, the biosynthesis of this activated form of sialic acid completely from endogenous metabolites is demonstrated for the first time in insect cells by expressing the mammalian genes required for the multistep conversion of endogenous UDP-GlcNAc to CMP-Neu5Ac. The genes for UDP-GlcNAc-2 epimerase/ManNAc kinase (EK), sialic acid 9-phosphate synthase (SAS), and CMP sialic acid synthetase (CSAS) were coexpressed in insect cells using baculovirus expression vectors, but the CMP-Neu5Ac and precursor Neu5Ac levels synthesized were found to be lower than those achieved with ManNAc supplementation due to feedback inhibition of the EK enzyme by CMP-Neu5Ac. When sialuria-like mutant EK genes, in which the site for feedback regulation has been mutated, were used, CMP Neu5Ac was synthesized at levels more than 4 times higher than that achieved with the wild-type EK and 2.5 times higher than that achieved with ManNAc feeding. Addition of N-acetylglucosamine (GlcNAc), a precursor for UDP-GlcNAc, to the media increased the levels of CMP-Neu5Ac even more to a level 7.5 times higher than that achieved with ManNAc supplementation, creating a bottleneck in the conversion of Neu5Ac to CMP-Neu5Ac at higher levels of UDP-GlcNAc. The present study provides a useful biochemical strategy to synthesize and enhance the levels of the sialylation donor molecule, CMP-Neu5Ac, a critical limiting substrate for the generation of complex glycoproteins in insect cells and other cell culture systems. PMID- 15895996 TI - Mechanistic insights on the folding of a large ribozyme during transcription. AB - RNA folding during transcription resembles folding in a cellular environment. We previously investigated the folding of a large ribozyme derived from a bacterial RNase P RNA during its transcription by the Escherichia coli RNA polymerase and the effect of the elongation factor NusA. We found that transcriptional pausing at a specific site induced by NusA significantly altered the folding pathway. In this work, we compare folding during transcription by the E. coli RNA polymerase of circularly permuted variants and site-specific mutants of the RNase P ribozyme to elucidate the molecular mechanism of transcriptional pausing and RNA folding. The effect of NusA-induced pausing depends on the order of RNA synthesis and only affects local folding of the RNA. Pausing likely prevents a misfolded structure between the 5' strand of a helix and its adjacent junction located in the specificity domain and a region known to bind single-stranded RNA located in the catalytic domain. These results lead to a structural model on how transcriptional pausing affects folding of RNase P RNA. Structural rearrangements of a nascent RNA transcript enhanced by transcriptional pausing may be a general feature of RNA folding during transcription. PMID- 15895997 TI - The D-methyl group in beta-lactamase evolution: evidence from the Y221G and GC1 mutants of the class C beta-lactamase of Enterobacter cloacae P99. AB - The beta-lactam antibiotics act through their inhibition of D-alanyl-D-alanine transpeptidases (DD-peptidases) that catalyze the last step of bacterial cell wall synthesis. Bacteria resist beta-lactams by a number of mechanisms, one of the more important of which is the production of beta-lactamases, enzymes that catalyze the hydrolysis of these antibiotics. The serine beta-lactamases are evolutionary descendants of DD-peptidases and retain much of their structure, particularly at the active site. Functionally, beta-lactamases differ from DD peptidases in being able to catalyze hydrolysis of acyl-enzyme intermediates derived from beta-lactams and being unable to efficiently catalyze acyl transfer reactions of D-alanyl-D-alanine terminating peptides. The class C beta-lactamase of Enterobacter cloacae P99 is closely similar in structure to the DD-peptidase of Streptomyces R61. Previous studies have demonstrated that the evolution of the beta-lactamase, presumably from an ancestral DD-peptidase similar to the R61 enzyme, included structural changes leading to rejection of the D-methyl substituent of the penultimate D-alanine residue of the DD-peptidase substrate. This seems to have been achieved by suitable placement of the side chain of Tyr 221 in the beta-lactamase. We show in this paper that mutation of this residue to Gly 221 produces an enzyme that more readily hydrolyzes and aminolyzes acyclic D alanyl substrates than glycyl analogues, in contrast to the wild-type beta lactamase; the mutant is therefore a more efficient DD-peptidase. Molecular modeling showed that the D-alanyl methyl group fits snugly into the space originally occupied by the Tyr 221 side chain and, in doing so, allows the bound substrate to assume a conformation similar to that on the R61 DD-peptidase, which has a hydrophobic pocket for this substituent. Another mutant of the P99 beta lactamase, the extended spectrum GC1 enzyme, also has space available for a D alanyl methyl group because of an extended omega loop. In this case, however, no enhancement of activity against D-alanyl substrates with respect to glycyl was observed. Accommodation of the penultimate D-alanyl methyl group is therefore necessary for efficient DD-peptidase activity, but not sufficient. PMID- 15895998 TI - A phosphoserine-lysine salt bridge within an alpha-helical peptide, the strongest alpha-helix side-chain interaction measured to date. AB - Phosphorylation is ubiquitous in control of protein activity, yet its effects on protein structure are poorly understood. Here we investigate the effect of serine phosphorylation in the interior of an alpha-helix when a salt bridge is present between the phosphate group and a positively charged side chain (in this case lysine) at i,i + 4 spacing. The stabilization of the helix is considerable and can overcome the intrinsically low preference of phosphoserine for the interior of the helix. The effect is pH dependent, as both the lysine and phosphate groups are titratable, and so calculations are given for several charge combinations. These results, with our previous work, highlight the different, context-dependent effects of phosphorylation in the alpha-helix. The interaction between the phosphate(2)(-) group and the lysine side chain is the strongest yet recorded in helix-coil studies. The results are of interest both in de novo design of peptides and in understanding the structural modes of control by phosphorylation. PMID- 15895999 TI - The quorum-quenching lactonase from Bacillus thuringiensis is a metalloprotein. AB - Lactonases from Bacillus species hydrolyze the N-acylhomoserine lactone (AHL) signaling molecules used in quorum-sensing pathways of many Gram-negative bacteria, including Pseudomonas aeruginosa and Erwinia carotovora, both significant pathogens. Because of sequence similarity, these AHL lactonases have been assigned to the metallo-beta-lactamase superfamily of proteins, which includes metalloenzymes of diverse activity, mechanism, and metal content. However, a recent study claims that AHL lactonase from Bacillus sp. 240B1 is not a metalloprotein [Wang, L. H., et al. (2004) J. Biol. Chem. 279, 13645]. Here, the gene for an AHL lactonase from Bacillus thuringiensis is cloned, and the protein is expressed, purified, and found to bind 2 equiv of zinc. The metal bound form of AHL lactonase catalyzes the hydrolysis of N-hexanoyl-(S)-homoserine lactone but not the (R) enantiomer. Removal of both zinc ions results in loss of activity, and reconstitution with zinc restores activity, indicating the importance of metal ions for catalytic activity. Metal content, sequence alignments, and X-ray absorption spectroscopy of the zinc-containing lactonase all support a proposed dinuclear zinc binding site similar to that found in glyoxalase II. PMID- 15896000 TI - Integration of the thylakoid membrane protein cytochrome b6 in the cytoplasmic membrane of Escherichia coli. AB - An overexpression system for spinach apocytochrome b(6) as a fusion protein to a maltose-binding protein in Escherichia coli was established using the expression vector pMalp2. The fusion of the cytochrome b(6) to the periplasmic maltose binding protein directs the cytochrome on the Sec-dependent pathway. The cytochrome b(6) has a native structure in the bacterial cytoplasmic membrane with both NH(2) and COOH termini on the same, periplasmic side of the membrane but has the opposite orientation compared to that in thylakoid. Our data also show that in the E. coli cytoplasmic membrane, apocytochrome b(6) and exogenic hemes added into a culture media spontaneously form a complex with similar spectroscopic properties to native cytochrome b(6). Reconstituted membrane-bound cytochrome b(6) contain two b hemes (alpha band, 563 nm; average E(m,7) = -61 +/- 0.84 and 171 +/- 1.27 mV). PMID- 15896001 TI - The receptor binding domain of apolipoprotein E is responsible for its antioxidant activity. AB - Apolipoprotein E (apoE) is a 34-kDa lipid-associated protein present in plasma and in the central nervous system. Previous studies have demonstrated that apoE has multiple functions, including the ability to transport lipids, regulate cell homeostasis, and inhibit lipid oxidation. The lipid binding domain of apoE has been localized to the carboxyl-terminal domain, whereas a cluster of basic amino acid residues within the N-terminal domain is responsible for its receptor binding activity. This study was undertaken to identify the domain in apoE responsible for its antioxidant activity. Results showed that apoE inhibits Cu(2+)-induced LDL oxidation by delaying conjugated diene formation in a concentration-dependent manner. Reductive methylation of lysine residues or cyclohexanedione modification of arginine residues in apoE abolished its ability to inhibit LDL oxidation. Additional studies showed that a 22-kDa peptide containing the N-terminal domain of apoE3 was more effective than a similar peptide with the apoE4 sequence in inhibiting Cu(2+)-induced LDL oxidation. In contrast, the 10-kDa peptide that contains the C-terminal domain of apoE was ineffective. Inhibition of Cu(2+)-induced LDL oxidation can also be accomplished with a peptide containing either a single sequence or a tandem repeat sequence of the receptor binding domain (residues 141-155) of apoE. Taken together, these results localized the antioxidant domain of apoE to its receptor binding domain and the basic amino acids in this domain are important for its antioxidant activity. PMID- 15896002 TI - Development of small designer aldolase enzymes: catalytic activity, folding, and substrate specificity. AB - Small (24-35 amino acid residues) peptides that catalyze carbon-carbon bond transformations including aldol, retro-aldol, and Michael reactions in aqueous buffer via an enamine mechanism have been developed. Peptide phage libraries were created by appending six randomized amino acid residues to the C-terminus or to the N-terminus of an 18-mer alpha-helix peptide containing lysine residues. Reaction-based selection with 1,3-diketones was performed to trap the amino groups of reactive lysine residues that were necessary for the catalysis via an enamine mechanism by formation of stable enaminones. The selected 24-mer peptides catalyzed the reactions with improved activities. The improved activities were correlated with improved folded states of the peptides. The catalyst was then improved with respect to substrate specificity by appending a phage display derived substrate-binding module. The resulting 35-mer peptide functioned with a significant proportion of the catalytic proficiency of larger protein catalysts. These results indicate that small designer enzymes with good rate acceleration and excellent substrate specificity can be created by combination of design and reaction-based selection from libraries. PMID- 15896003 TI - Calcium-binding sites of calmodulin and electron transfer by inducible nitric oxide synthase. AB - Like that of the neuronal nitric oxide synthase (nNOS), the binding of Ca(2+) bound calmodulin (CaM) also regulates the activity of the inducible isoform (iNOS). However, the role of each of the four Ca(2+)-binding sites of CaM in the activity of iNOS is unclear. Using a series of single-point mutants of Drosophila melanogaster CaM, the effect that mutating each of the Ca(2+)-binding sites plays in the transfer of electrons within iNOS has been examined. The same Glu (E) to Gln (Q) mutant series of CaM used previously [Stevens-Truss, R., Beckingham, K., and Marletta, M. A. (1997) Biochemistry 36, 12337-12345] to study the role of the Ca(2+)-binding sites in the activity of nNOS was used for these studies. We demonstrate here that activity of iNOS is dependent on Ca(2+) being bound to sites II (B2Q) and III (B3Q) of CaM. Nitric oxide ((*)NO) producing activity (as measured using the hemoglobin assay) of iNOS bound to the B2Q and B3Q CaMs was found to be 41 and 43% of the wild-type activity, respectively. The site I (B1Q) and site IV (B4Q) CaM mutants only minimally affected (*)NO production (95 and 90% of wild-type activity, respectively). These results suggest that NOS isoforms, although all possessing a prototypical CaM binding sequence and requiring CaM for activity, interact with CaM differently. Moreover, iNOS activation by CaM, like nNOS, is not dependent on Ca(2+) being bound to all four Ca(2+)-binding sites, but has specific and distinct requirements. This novel information, in addition to helping us understand NOS, should aid in our understanding of CaM target activation. PMID- 15896006 TI - Microfluidics: from dynamic lattices to periodic arrays of polymer disks. AB - We used coupling of flow and geometric confinement to assemble emulsion droplets in two-dimensional gliding lattices with a high degree of order and symmetry. Highly monodisperse discoid droplets with circular shapes were generated in a microfluidic flow-focusing device. Originally, close-packed lattices formed from these circular discoid droplets. Progressive confinement led to the gradual deformation of the circular disks: first, they elongated in the direction parallel to the direction of flow and then transformed into hexagons. Assembly driven by the combination of flow and confinement also allowed for the formation of lattices from droplets with a bimodal size distribution. We used photopolymerization of the monomer droplets to trap the lattice structure in the solid state and produce highly periodic arrays of solid polymer disks. PMID- 15896007 TI - Sensing DNA hybridization via ionic conductance through a nanoporous electrode. AB - We show that nanoporous alumina modified with covalently linked DNA can be used to detect target DNA by monitoring the increase in impedance at the electrode upon DNA hybridization, which resulted from blocking the pores to ionic flow. Using cyclic voltammetry, direct current conductance, and impedance spectroscopy we confirm the importance of pore size: the effect is observed with 20-nm diameter pores and is absent for 200-nm pores. PMID- 15896008 TI - Scanning tunneling microscope imaging of (CH3S)2 on Cu(111). AB - Scanning tunneling microscope (STM) images of isolated molecules of dimethyl disulfide, (CH(3)S)(2), adsorbed on the Cu(111) surface were successfully obtained at a sample temperature of 4.7 K. A (CH(3)S)(2) molecule appears as an elliptic protrusion in the STM images. From density functional theory calculation, it was suggested that the bright part in the protrusion corresponds to the molecular orbital which is widely spread around H atoms in each CH(3) group in the (CH(3)S)(2) molecule. The STM images revealed that the molecules have a total of six equivalent adsorption orientations on Cu(111), which are given by the combination of three equivalent adsorption sites and two conformational isomers for each adsorption site. PMID- 15896009 TI - Perfecting imperfect "monolayers": removal of siloxane multilayers by CO2 snow treatment. AB - Self-assembled monolayers (SAMs) of N-(3-triethoxysilylpropyl)-4 hydroxybutyramide were prepared on silicon oxide on silicon (Si/SiO(2)). Initial silane adsorption and high-temperature annealing led to a stable base monolayer with many large over-lying islands of disordered multilayers as a result of the non-self-limited growth process. The disordered multilayers were hydrolyzed and subsequently removed by CO(2) snow treatment. The resulting films were one monolayer thick as measured by ellipsometry. Atomic force microscopy, attenuated total reflection Fourier transform infrared spectroscopy, and contact angle analysis showed that the films were composed of monolayers with full and uniform surface coverage rather than nonuniform coverage by islands or patches of multilayers. Monolayers of octadecyltrichlorosilane were also prepared by multilayer removal via CO(2) treatment, showing the general applicability of the technique toward siloxane SAMs. We believe that CO(2) is an excellent solvent for weakly bound and hydrolyzed molecules that compose multilayers, and this ability to prepare near-perfect monolayer films from imperfect ones allows for less stringent formation conditions. PMID- 15896010 TI - Dynamic self-assembly of polymer colloids to form linear patterns. AB - A dynamic self-assembling process is reported which involves drying a droplet of positively charged colloidal suspension on a flat negatively charged hydrophilic surface. This extremely simple method affords lines of colloidal particles with regular 1.5-4.5 microm line spacing and smaller than 2 microm line width over a broad surface area. The ordered region diffracts light to display an iridescent appearance and generates first-order diffraction spots when illuminated by a He Ne laser. A periodic stick-slip motion of the drying liquid front is observed during the drying process using optical microscopy. The periodic motion must be related to the periodic particle deposition. We propose that the simultaneous deposition of the particles at a fixed distance (i.e., the line spacing) behind the previous line of particles where the contact line is pinned is in turn responsible for the periodic stick-slip motion. The key distinguishing feature of the present system is the attractive interaction between the particles and the surface, which instigates the periodicity of the particle deposition. PMID- 15896011 TI - Evidence of surface charge at the air/water interface from thin-film studies on polyelectrolyte-coated substrates. AB - The stability of thin water films on silicon substrates coated with cationic and anionic polyelectrolytes was investigated by the thin film pressure balance technique. Depending on the surface charge of the substrate, the water films are either stable (on negatively charged wafers) or rupture rapidly (on positively charged wafers). It is supposed that this behavior is due to a negative surface charge of the free water surface. The underlying assumption that the films' stability is due to electrostatic interactions is supported by measurements of the disjoining pressure on silicon wafers with a native oxide layer, which indicates a decrease of the film thickness, and thus decreasing repulsive interaction between the two film interfaces, with increasing ionic strength. PMID- 15896012 TI - Thermally stable holographic surface relief gratings and switchable optical anisotropy in films of an azobenzene-containing polyelectrolyte. AB - In a search for effective polymer film material for holographic surface patterning, commercially available azobenzene polyelectrolyte has been employed. Films of good optical quality in a wide range of thickness were produced. Optical dichroism up to 0.19 was induced upon irradiation with linearly polarized light. Surface relief gratings with amplitudes up to 630 nm and diffraction efficiency of 37% were inscribed by holographic exposure to the light of 488 nm. Due to the ionic nature of the material, the relief was stable at least up to the temperature of decomposition (ca. 200 degrees C) but could be erased and inscribed again by light. PMID- 15896013 TI - Electrochemical functionalization of single-walled carbon nanotubes in large quantities at a room-temperature ionic liquid supported three-dimensional network electrode. AB - Electrochemical functionalization of single-walled carbon nanotubes (SWNTs) was one of selective, clean, and nondestructive chemical methods. But in previous studies it met difficulties in homogeneous electrografting of SWNTs in large quantities because the reaction was often localized on a very thin film (ca. 2 microm). In this report, a room-temperature ionic liquid (RTIL) supported three dimensional network SWNT electrode was first utilized to break through this barrier. In this work, large quantities of SWNTs were considerably untangled in RTILs so as to greatly increase the effective area of the electrode. N succinimidyl acrylate (NSA), as a model monomer, was dissolved in the supporting RTILs and was electrografted onto SWNTs (SWNTs-poly-NSA). As an application example, glucose oxidase was directly covalently anchored on the SWNTs-poly-NSA assembly, and the electrocatalytic oxidation of glucose in this assembly was investigated. RTILs opened a new path in electrochemical functionalization of SWNTs. PMID- 15896014 TI - Deposition of amorphous calcium carbonate hemispheres on substrates. AB - The amorphous calcium carbonate (ACC) hemispheres were deposited on the mica and poly(diallyldimethylammonium chloride) modified surface. The form of the ACC deposit on the substrates can be controlled by modifying the substrate surface, the introduction of additives, or both. It demonstrated that substrates (insoluble matrix) and additives (soluble macromolecules) have significant influence on the crystallization of CaCO(3). PMID- 15896015 TI - Three-dimensional crystal alignment using a time-dependent elliptic magnetic field. AB - A theoretical study has been presented to show that it is possible to align three different diamagnetic susceptibility axes (chi(3) < chi(2) < chi(1) < 0) of a crystallite with respect to the laboratory coordinate system (x, y, z). The time dependent magnetic field that periodically changes in direction and intensity on the xy plane in an elliptic manner (the intensity stronger in the x direction) at a rate quicker than the intrinsic rate of magnetic response causes the three dimensional alignment, that is, chi(1) parallel x, chi(2) parallel y, and chi(3) parallel z. The fluctuation of the three susceptibility axes around the corresponding laboratory coordinates is estimated in terms of the fluctuation around the minimum of the time-averaged magnetic potential. This technique enables the three-dimensional alignment of the crystallographic axes. PMID- 15896016 TI - Construction of a tethered poly(ethylene glycol) surface gradient for studies of cell adhesion kinetics. AB - Surface gradients can be used to perform a wide range of functions and represent a novel experimental platform for combinatorial discovery and analysis. In this work, a gradient in the coverage of a surface-immobilized poly(ethylene glycol) (PEG) layer is constructed to interrogate cell adhesion on a solid surface. Variation of surface coverage is achieved by controlled transport of a reactive PEG precursor from a point source through a hydrated gel. Immobilization of PEG is achieved by covalent attachment of the PEG molecule via direct coupling chemistry to a cystamine self-assembled monolayer on gold. This represents a simple method for creating spatial gradients in surface chemistry that does not require special instrumentation or microfabrication procedures. The structure and spatial distribution of the PEG gradient are evaluated via ellipsometry and atomic force microscopy. A cell adhesion assay using bovine arteriole endothelium cells is used to study the influence of PEG thickness and chain density on biocompatibility. The kinetics of cell adhesion are quantified as a function of the thickness of the PEG layer. Results depict a surface in which the variation in layer thickness along the PEG gradient strongly modifies the biological response. PMID- 15896017 TI - Nanoscale functionalization and site-specific assembly of colloids by particle lithography. AB - Numerous studies have demonstrated the bottom-up assembly of complex structures such as colloidal crystals, close-packed aggregates, and even rings and tetramers. In this paper we produce a simple localized and nanoscale charge distribution on the surfaces of individual colloidal microspheres using our technique of "particle lithography". In this technique parts of the microspheres are masked off, while polyelectrolytes (or other molecules) cover the remaining portions of the microspheres. The effectiveness of this process is demonstrated by the accurate and reproducible production of colloidal heterodoublets composed of oppositely charged microspheres. These "colloidal molecules" have the potential for significantly higher information content than previous attempts in the literature. The particle lithography technique is advantageous because it is not limited by the resolution of photolithography or by functionalizing chemistries, and the technique opens the door for complex site-specific functionalization of particles. PMID- 15896018 TI - Chemical functionalization of silica and alumina particles for dispersion in carbon dioxide. AB - The steric stabilization and flocculation of modified silica and alumina particle suspensions in condensed CO(2) were studied. Silica particles (average diameters of 7 and 12 nm) were functionalized using chlorosilanes of the form C(n)F(2n+1)CH(2)CH(2)Si(CH(3))(2)Cl (n = 8, 4, or 1) to give C(n)F(2n+1)-silica. Alumina particles (diameter of 8-14 nm) were grafted with C(8)F(17)CH(2)CH(2)Si(OEt)(3) and chemically modified with perfluorononanoic acid to yield C(8)F(17)-alumina and C(8)F(17)COOH-alumina, respectively. Elemental analysis and thermogravimetric analysis on the derivatized particles were carried out, and surface coverage was calculated. The stabilization of these modified particles in condensed CO(2) was quantified using turbidimetry. Particle stability was found to increase with increasing fluorinated tail length, temperature, and CO(2) density. Unmodified particles and those modified with only -CF(3) tails were unstable in condensed CO(2). Stabilization in supercritical CO(2) is continuous up to 24 h for the C(n)F(2n+1)-silica (n >/= 4) particles and 96 h for the C(8)F(17)-alumina particles. The C(8)F(17)COOH-alumina particles gave a significantly higher graft density than the C(8)F(17)-alumina particles but are not as stable in CO(2). The C(8)F(17)-alumina particles were stable at lower CO(2) densities than the modified silica particles. This stability difference may be attributed to the precursor organosilanes being monofunctional (modified silica) versus trifunctional (modified alumina), producing different structures on the surface. PMID- 15896019 TI - Effect of n-alkanes on asphaltene structuring in petroleum oils. AB - The interactions between asphaltenes and short- to medium-chain n-alkanes were studied using titration microcalorimetry and inverse chromatography. The exothermic heat effects observed upon mixing of asphaltenes and n-alkanes were interpreted in terms of assembling of the two types of compounds into mixed structures. We show that the energy of the interactions between n-alkanes and the asphaltene hydrocarbon chains is close to the energy of the interactions between the asphaltene chains. We propose that the latter interactions are responsible for the formation of the asphaltene aggregates and are the driving force of the aggregate assembly into higher structures. PMID- 15896020 TI - Formation of intra- and interparticle polyelectrolyte complexes between cationic nanogel and strong polyanion. AB - Polyelectrolyte complex formation of a strong polyanion, potassium poly(vinyl alcohol) sulfate (KPVS), with positively charged nanogels was studied at 25 degrees C in aqueous solutions with different KCl concentrations (C(s)) as a function of the polyion-nanogel mixing ratio based on moles of anions versus cations. Used as the gel sample was a polyampholytic nanogel consisting of lightly cross-linked terpolymer chains of N-isopropylacrylamide, acrylic acid, and 1-vinylimidazole; thus, the complexation was performed at pH 3 at which the imidazole groups are fully protonated to generate positive charges. Turbidimetric titration was employed to vary the mixing ratio. Also employed for studies of the resulting complexes at different stages of the titration were dynamic light scattering (DLS) and static light scattering (SLS) techniques. It was found from the titration as well as DLS and SLS that there is a critical mixing ratio (cmr) at which both the size and molar mass of the complexed gel particles abruptly increase. The value of the cmr at C(s) = 0 or 0.01 M (mol/L) was observed at approximately 1:1 mixing ratio of anions versus cations but at lower mixing ratios than the 1:1 ratio under conditions of C(s) = 0.05 and 0.1 M. At the mixing ratios less than the cmr, the molar mass of the complex agrees with that of one gel particle with the calculated amount of the bound KPVS ions, indicating the formation of an "intraparticle" KPVS-nanogel complex, by the aggregation of which an "interparticle" complex is formed at the cmr. During the process of the intraparticle complex formation, both the hydrodynamic radius by DLS and the radius gyration by SLS decreased with increasing mixing ratio, demonstrating the gel collapse due to the complexation. At C(s) = 0 or 0.01 M and under conditions where the amount of KPVS bindings was less than half of the nanogel cations, however, the decrease of the hydrodynamic radius was very small, while the radius gyration fell monotonically. These results were discussed in connection with a collapse of dangling chains attached to the nanogel surface by the binding of KPVS. PMID- 15896021 TI - Emulsion polymerization of styrene and methyl methacrylate using a hydrophobically modified inulin and comparison with other surfactants. AB - The use of a new class of graft polymer surfactants, based on inulin, in emulsion polymerization of poly(methyl methacrylate) (PMMA) and polystyrene (PS) particles is described. PS and PMMA were synthesized by emulsion polymerization, and stable particles with a high monomer content (50 wt %) were obtained with a very small amount of polymeric surfactant ([surfactant]/[monomer] = 0.0033). The latex dispersions were characterized by dynamic light scattering and by transmission electron microscopy to obtain the average particle size and the polydispersity index, and the stability was determined by turbidimetry measurements and expressed in terms of critical coagulation concentration. The last section gives a comparison of PMMA particles prepared by emulsion polymerization using classical surfactants from different types as emulsifiers with that obtained using the copolymer surfactant. It shows the superiority of INUTEC SP1 as it is the only one that allows stable particles at 20 wt % monomer content, with a smaller ratio [surfactant]/[monomer] = 0.002. PMID- 15896022 TI - Effects of electrolyte concentration and pH on the coalescence stability of beta lactoglobulin emulsions: experiment and interpretation. AB - Experimental results are presented about the effects of ionic strength and pH on the mean drop-size after emulsification and on the coalescence stability of emulsions, stabilized by a globular protein beta-lactoglobulin (BLG). The mean drop-size is determined by optical microscopy, whereas the coalescence stability is characterized by centrifugation. In parallel experiments, the zeta-potential and protein adsorption on drop surface are determined. The experiments are performed at two different BLG concentrations, 0.02 and 0.1 wt%. The electrolyte concentration in the aqueous phase, C(EL), is varied between 1.5 mM and 1 M, and pH is varied between 4.0 and 7.0. The experiments show that the mean drop-size after emulsification depends slightly on C(EL), at fixed protein concentration and natural pH = 6.2. When pH is varied, the mean drop-size passes through a maximum at fixed protein and electrolyte concentrations. A monolayer protein adsorption is registered in the studied ranges of C(EL) and pH at low BLG concentration of 0.02 wt%. In contrast, a protein multilayer is formed at higher BLG concentration, 0.1 wt%, above a certain electrolyte concentration (C(EL) > 100 mM, natural pH). The experimental results for the emulsion coalescence stability are analyzed by considering the surface forces acting between the emulsion drops. The electrostatic, van der Waals, and steric interactions are taken into account to calculate the barriers in the disjoining pressure isotherm at the various experimental conditions studied. The comparison of the theoretically calculated and the experimentally determined coalescence barriers shows that three qualitatively different cases can be distinguished. (1) Electrostatically stabilized emulsions, with monolayer protein adsorption, whose stability can be described by the DLVO theory. (2) Sterically stabilized emulsions, in which the drop-drop repulsion is created mainly by overlapping protein adsorption multilayers. A simple theoretical model is shown to describe emulsion stability in these systems. (3) Sterically stabilized emulsions with a monolayer adsorption on drop surface. PMID- 15896023 TI - Structure of a hydrophilic-hydrophobic block copolymer and its interactions with salt and an anionic surfactant. AB - The aggregation of a hydrophilic-hydrophobic diblock copolymer consisting of poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) and poly(methyl methacrylate) (PMMA) in aqueous solution has been investigated by small-angle neutron scattering. This polybase is extensively protonated at low pH and forms micelles with a dense core of PMMA and a diffuse coronal layer of cationic PDMAEMA. Addition of salt induced micellar growth, brought about by charge screening and more efficient packing of the chains. As a result, the aggregation number increased from 8 up to 31. A similar effect was observed at low concentrations of an anionic surfactant, sodium dodecyl sulfate (SDS) since the net cationic charge in the hydrophilic coronal layer was reduced due to surfactant binding. However, at higher surfactant concentrations, a drastic structural reorganization occurred, as the PMMA became solubilized into the SDS micellar cores and the PDMAEMA chains interacted with the surfactant micelles, resulting in a "pearl necklace" structure. The presence of the cationic polyelectrolyte significantly increased the population of SDS micelles by effectively lowering the critical micelle concentration of this anionic surfactant. PMID- 15896024 TI - Solubilization of phenols in anionic polyelectrolyte gels with adsorbed cationic surfactant. AB - Solubilization isotherms for various phenols in cetylpyridinium chloride (CPC) polyelectrolyte gel aggregates have been determined in order to compare solubilization within these aggregates with that in free micelles and to examine the effects of gel chemistry and structure on solubilization. The isotherms describing solubilization are quite similar to those found for free surfactant in solution. Solutes that are more hydrophobic give rise to larger solubilization constants with trends similar to what is seen for hydrophobic effects in adsorption from aqueous solutions onto hydrophobic solids. The solubilization constants decrease as the fraction of solute in the aggregates increases, indicating that the solutes partition into the palisade region of the aggregates. Solubilization is found to be quite insensitive to changes in gel structure (cross-linker varying from 1% to 3%) and chemistry (poly(acrylic acid) versus poly(methacrylic acid) and neutralization from 50% to 100%). However, the switch from poly(acrylic acid) to poly(methacrylic acid) did give rise to a slight decrease in magnitude of the slope of the isotherm. The most significant factors appear to be the initial concentration of surfactant in solution and the ratio of surfactant solution to gel amount. A decrease in surfactant concentration (especially combined with an increase in solution volume) gives rise to a decrease in solubilization constants. PMID- 15896025 TI - Aggregate formation of binary nonionic surfactant mixtures on hydrophilic surfaces. AB - Adsorption of surfactant mixtures on hydrophilic solid surfaces is of considerable theoretical and practical importance. Cooperative adsorption of nonionic surfactant mixtures of nonyl phenol ethoxylated decyl ether (NP-10) and n-dodecyl-beta-d-maltoside (DM) on silica and alumina was investigated in this study with a view to elucidate the nanostructures of their aggregates. In the mixed system, DM is identified to be the "active" component and NP-10 is the "passive" one for the process of adsorption on alumina, while their roles are reversed for silica. The difference in the adsorptive interactions of the surfactants with the above minerals is attributed to the differences in the molecular structures of the surfactants. To better understand the interaction between surfactants at solid/solution interface from a molecular structure point of view, the nanostructures of mixed surface aggregates have been quantitatively predicted for the first time using a modified packing parameter: the structures are spherical or cylindrical on silica and those on alumina undergo a spherical to-cylindrical-to-bilayer transition with the addition of the active component. This work offers a new way for developing of structure-performance relationships. PMID- 15896026 TI - Assembly of colloidal particles into microwires using an alternating electric field. AB - We have investigated the dielectrophoretic assembly of colloidal gold, carbon black, and carbon nanotubes into electrical wires. The resulting microwires have diameters less than 1 microm, with lengths ranging from 5 microm to 3 mm. Current voltage curves for these wires indicate an ohmic response, where the resistance is determined by the type of colloid and by the frequency of the alternating field used to grow the wires. The predicted frequency dependence of dielectrophoresis is confirmed by experiment. Measurements of the threshold voltage for initial wire growth are also presented. These experiments demonstrate that a variety of nanoparticles can be assembled into microwires for sensor applications. PMID- 15896027 TI - Charge stabilization in nonpolar solvents. AB - While the important role of electrostatic interactions in aqueous colloidal suspensions is widely known and reasonably well-understood, their relevance to nonpolar suspensions remains mysterious. We measure the interaction potentials of colloidal particles in a nonpolar solvent with reverse micelles. We find surprisingly strong electrostatic interactions characterized by surface potentials, |ezeta|, from 2.0 to 4.4 k(B)T and screening lengths, kappa(-1), from 0.2 to 1.4 microm. Interactions depend on the concentration of reverse micelles and the degree of confinement. Furthermore, when the particles are weakly confined, the values of |ezeta| and kappa extracted from interaction measurements are consistent with bulk measurements of conductivity and electrophoretic mobility. A simple thermodynamic model, relating the structure of the micelles to the equilibrium ionic strength, is in good agreement with both conductivity and interaction measurements. Since dissociated ions are solubilized by reverse micelles, the entropic incentive to charge a particle surface is qualitatively changed from aqueous systems, and surface entropy plays an important role. PMID- 15896028 TI - Inhibition of the beta-cyclodextrin catalyzed dediazoniation of 4 nitrobenzenediazonium tetrafluoroborate. blocking effect of sodium dodecyl sulfate. AB - We have investigated the effects of sodium dodecyl sulfate, SDS, on the reaction between 4-nitrobenzenediazonium, 4NBD, ions and beta-cyclodextrin, beta-CD, under acidic conditions at T = 60 degrees C by employing a combination of spectrophotometric, chromatographic, and conductometric techniques. Previous studies under acidic conditions indicate that the secondary -OH groups of beta-CD solvate 4NBD ions, which are included in the beta-CD cavity, leading to the formation of a highly unstable transient diazo ether complex that undergoes homolytic fragmentation with an observed rate constant about 1700 times higher than that in pure aqueous acid solution (t(1/2) = 6 h at T = 60 degrees C) when [beta-CD]/[4NBD] = 40. Addition of SDS to a 4NBD/beta-CD system makes the k(obs) values decrease up to its value in a SDS micellar solution, which is similar to that in aqueous acid solution. Dediazoniation product distribution is significantly affected; the reaction between 4NBD and beta-CD ([beta-CD]/[4NBD] = 40), in the absence of SDS, proceeds exclusively through a homolytic mechanism leading to the quantitative formation of nitrobenzene, ArH, but addition of SDS turns over the mechanism by promoting the heterolytic mechanism. In addition, mixtures of 4-nitrophenol, ArOH, and ArH dediazoniation products are formed; their relative yields depend on the amount of added SDS so that at very high [SDS(T)], the heterolytic mechanism becomes the predominant one. Results are consistent with conductometric measurements showing that addition of beta-CD to an aqueous surfactant solution inhibits micelle formation and elevates CMC(app) values because CD encapsulation of surfactant monomers competes with the micellization process and are interpreted in terms of SDS monomers blocking the beta-CD cavity by forming a nonreactive complex, releasing 4NBD to the bulk solution. PMID- 15896029 TI - Quasi-static electrorheological properties of hematite/silicone oil suspensions under DC electric fields. AB - In this work, a modified rheometer has been used to gain information on the "start-up" of the shear flow of an electrorheological (ER) fluid consisting of hematite particles dispersed in silicone oil. The results show that unelectrified suspensions behave essentially as fluids, continuously deforming upon application of shear. However, this behavior changes in the presence of an electric field. For low fields and low volume fractions of solids, a solidlike (drastic increase in shear stress after the strain is applied) behavior is observed for small deformations. If the strain is increased, the yield starts and a transition to a viscoelastic-plastic nature is observed. Finally, a plastic behavior is characteristic of the post-yield regime. If the field strength and solids content are high, a discontinuous flow profile develops. These results, together with direct structural observations, suggest that the observed behavior is compatible with the formation of layers of particles electrophoretically deposited on the electrodes; the layers turn into rings when the shear field is applied. It is the slip of the fluid between these rings that can be considered responsible for the ER effect in these suspensions. PMID- 15896030 TI - Temperature-induced growth of wormlike copolymer micelles. AB - We study the temperature-induced growth of polymer micelles based on Pluronic P84 in brine (2 M NaCl) using small-angle neutron scattering, static and dynamic light scattering, and viscometry as a function of temperature and polymer concentration. Spherical micelles below 30 degrees C are shown to grow between about 30 and 40 degrees C into wormlike micelles long enough to enter the semidilute regime for polymer volume fraction larger than 0.005. The entanglements in this regime are responsible for a huge increase in the viscosity. Above about 41 degrees C, the micellar aggregates become denser as the cloud point is approached and the viscosity drops. PMID- 15896031 TI - Influence of surfactant on gas bubble stability. AB - Gas-bubble stability is achieved either by a reduction in the Laplace pressure or by a reduction in the permeability of the gas-liquid interface. Although insoluble surfactants have been shown definitively in many studies to lower the permeability of the gas-liquid interface and hence increase the resistance to interfacial mass transfer, remarkably little work has been done on the effects of soluble surfactants. An experimental system was developed to measure the effect of the soluble surfactant dodecyl trimethylammonium bromide on the desorption and absorption of carbon dioxide gas through a quiescent planar interface. The desorption experiments conformed to the model of non-steady-state molecular diffusion. The absorption experiments, however, produced an unexpected mass transfer mechanism, with surface renewal, probably because of instability in the density gradient formed by the carbon dioxide. In general, the soluble surfactant produced no measurable reduction in the rate of interfacial mass transfer for desorption or absorption. This finding is consistent with the conclusion of Caskey and Barlage that soluble surfactants produce a significantly lower resistance to interfacial mass transfer than do insoluble surfactants. The dynamic adsorption and desorption of the surfactant molecules at the gas-liquid interface creates short-term vacancies, which presumably permit the unrestricted transfer of the gas molecules through the interface. This surfactant exchange does not occur for insoluble surfactants. Gas bubbles formed in the presence of a high concentration of soluble surfactant were observed to dissolve completely, while those formed in the presence of the insoluble surfactant stearic acid did not dissolve easily, and persisted for very long periods. The interfacial concentration of stearic acid rises during bubble dissolution, as it is insoluble, and must eventually achieve full monolayer coverage and a state of compression, lowering the permeability of the interface. Thus, insoluble surfactants or hydrophobic impurities from solid surfaces may account for increased bubble stability. PMID- 15896032 TI - Formation of self-supporting reversible cellular networks in suspensions of colloids and liquid crystals. AB - In mixtures of thermotropic liquid crystals with spherical poly (methyl methacrylate) particles, self-supporting networklike structures are formed during slow cooling past the isotropic-to-nematic phase transformation. To characterize the process of network formation in terms of morphology, phase transformation kinetics, and mechanical properties, we have combined data from polarization and laser scanning confocal microscopy with calorimetric, NMR, and rheological results. Our data suggest that the mechanism of network formation is dominated by a broadened temperature and time interval of phase transformation rather than by particle size or concentration. The observation that the width of the transformation interval strongly depends on sample preparation supports the hypothesis that a third component, most likely alkane remnants slowly liberated from the particles, plays a crucial role. In addition, calorimetric findings for liquid crystal/colloid mixtures, heated and cooled up to 13 times, point to separation of the liquid crystal into two compartments with different phase transformation kinetics. This could be explained by redistribution and enrichment of alkane in the particle-composed network walls. A further increase of the storage modulus, G', and incomplete dissolution of the networks in the isotropic state indicate that the formation of two compartments during repeated temperature cycles stabilizes the network and confers strong memory effects. PMID- 15896033 TI - Hexagonal liquid crystalline phases formed in ternary systems of Brij 97-water ionic liquids. AB - Phase diagrams of two ionic liquids: hydrophobic 1-butyl-3-methylimidazolium hexafluorophosphate (bmim-PF(6)) and relatively hydrophilic 1-butyl-3 methylimidazolium tetrafluoroborate (bmim-BF(4)) in aqueous solutions of Brij 97 were determined at 25 degrees C. Two hexagonal liquid crystalline phases formed in bmim-PF(6)- and bmim-BF(4)-containing ternary systems were investigated by means of small-angle X-ray scattering (SAXS) and rheological techniques, with comparison of composition and temperature effects. From analysis of the SAXS data, bmim-PF(6) is dominantly penetrated between the oxyethylene chains of surfactant molecules, whereas bmim-BF(4) is mainly located in the water layer of hexagonal phases. The strength of the network of hexagonal phase formed in the Brij 97/water/bmim-BF(4) system is appreciably stronger than that of the Brij 97/water/bmim-PF(6) system, indicated by the smaller area of the surfactant molecule at the interface and the higher moduli (G', G' '). Temperature has a converse effect on the lattice parameters of the two hexagonal phases. PMID- 15896034 TI - Cracking in drying latex films. AB - Thin films of latex dispersions containing particles of high glass transition temperature generally crack while drying under ambient conditions. Experiments with particles of varying radii focused on conditions for which capillary stresses normal to the film deform the particles elastically and generate tensile stresses in the plane of the film. Irrespective of the particle size, the drying film contained, simultaneously, domains consisting of a fluid dispersion, a fully dried packing of deformed spheres, and a close packed array saturated with water. Interestingly, films cast from dispersions containing 95-nm sized particles developed tensile stresses and ultimately became transparent even in the absence of water, indicating that van der Waals forces can deform the particles. Employing the stress-strain relation for a drying latex film along with the well known Griffith's energy balance concept, we calculate the critical stress at cracking and the accompanying crack spacing, in general agreement with the observed values. PMID- 15896035 TI - Light-directed movement of polymer microstructures. AB - Light has been used to induce photochemical changes in the surface chemistry of porous polymer microstructures giving rise to a substantial change in volume. When illumination is asymmetric, this results in light-directed motion of the structure. Swellable trimethylolpropane trimethacrylate cross-linked poly(2 hydroxylethyl methacrylate) microstructures were constructed by azo-bis isobutyronitrile photopolymerization using a 20 x 0.5 NA microscope objective and 365 nm laser excitation. Structures were aminiated with glycine and protected with the photolabile group 4-nitroveratryloxycarbanyl (NVOC). Addition of NVOC resulted in a volume increase >10% when performed in the solvent N,N' dimethylformamide. Photochemical cleavage of NVOC using asymmetric illumination of a cone-shaped microstructure with a 365 nm laser induced polymer shrinkage in excess of 4% at the base of the cone and resulted in a maximum velocity of 1 mm/s at the tip of the cone. Symmetric illumination gave rise to displacement of solvent from the microstructure due to shrinkage with a velocity in excess of 0.01 mm/s. This system could in principle be used for light-directed movement of micromechanical systems, optical control of microfluidics, or light activated chemical delivery. PMID- 15896036 TI - Influence of NaCl on the behavior of PEO-PPO-PEO triblock copolymers in solution, at interfaces, and in asymmetric liquid films. AB - The solution behavior of the polymeric surfactant Pluronic F127 (PEO(99)PPO(65)PEO(99)) and its adsorption behavior on aqueous-silica and aqueous air interfaces, as well as the disjoining pressure isotherms of asymmetric films (silica/aqueous film/air) containing F127, are studied. The interfacial properties of adsorbed F127 layers (the adsorbed amount Gamma and the thickness h) as well as the aqueous wetting film properties [film thickness (h) and refractive indexes] were studied via ellipsometry. The solution properties of F127 were investigated using surface tensiometry and light scattering. The interactions between the air-water and silica-water interfaces were measured with a thin film pressure balance technique (TFB) and interpreted in terms of disjoining pressure as a function of the film thickness. The relations between the behaviors of the asymmetric films, adsorption at aqueous air, and aqueous silica interfaces and the solution behavior of the polymeric surfactant are discussed. Special attention is paid to the influence of the concentrations of F127 and NaCl. Addition of electrolyte lowers the critical micelle concentration, diminishes adsorption on silica, and increases the thickness of the asymmetric film. PMID- 15896037 TI - Adsorption of formaldehyde on Pt(111) and Pt(100) electrodes: cyclic voltammetry and scanning tunneling microscopy. AB - The adsorption of formaldehyde (HCHO) on Pt(111) and Pt(100) electrodes was examined by cyclic voltammetry (CV) and in situ scanning tunneling microscopy (STM) in 0.1 M HClO(4). The extent of HCHO adsorption at both Pt electrodes was evaluated by comparing the CVs, particularly for the hydrogen adsorption and desorption between 0.05 and 0.4 V, obtained in 0.1 M HClO(4) with and without HCHO. The adsorption of HCHO on these Pt electrodes was significant only when [HCHO] >/= 10 mM. Adsorbed organic intermediate species acted as poisons, blocking Pt surfaces and causing delays in the oxidation of HCHO. Compared to Pt(111), Pt(100) was more prone to poisoning, as indicated by a 200 mV positive shift of the onset of HCHO oxidation. However, Pt(100) exhibited an activity 3 times higher than that of Pt(111), as indicated by the difference in peak current density of HCHO oxidation. Molecular resolution STM revealed highly ordered structures of Pt(111)-( radical7 x radical7)R19.1 degrees and Pt(100)-( radical2 x radical2) in the potential region between 0.1 and 0.3 V. Voltammetric measurements further showed that the organic poisons produced by HCHO adsorption behaved differently from the intentionally dosed CO admolecules, which supports the assumption for the formation of HCO or COH adspecies, rather than CO, as the poison. On both Pt electrodes, HCHO oxidation commenced preferentially at step sites at the onset potential of this reaction, but it occurred uniformly at the peak potentials. PMID- 15896038 TI - Template-directed assembly on an ordered microsphere array. AB - We investigated the capability of an ordered array of microspheres to act as a template for deposition and ordering of a subsequent layer of microspheres. An evaporation-based technique was used to deposit monolayers of large colloidal spheres. A novel technique for selective deposition of polyelectrolyte film was used to stabilize the arrays and optimize the bead-substrate interaction. The template behavior of face-centered cubic and body-centered cubic (bcc) microsphere arrays was studied by optical and scanning electron microscopy, and the packing geometry was found to have a dramatic effect on the arrangement of the subsequent layer. A geometrical interpretation of the experimental data explains why a bcc bead array is well suited to act as a template for an additional layer of microspheres. PMID- 15896039 TI - Probing the structure of supported membranes and tethered oligonucleotides by fluorescence interference contrast microscopy. AB - Fluorescence interference contrast microscopy (FLIC) is a powerful method to structurally characterize fluorescent objects with nanometer-scale resolution in the z direction. Here we use FLIC to characterize the water layer underlying supported membranes and membrane-tethered double-stranded oligonucleotides. FLIC measurements of supported membranes containing the lipid-anchored fluorescent dye DiI in both leaflets indicate the thickness of the water layer separating the solid support and the lower lipid leaflet is 1.3 +/- 0.2 nm. Addition of cobalt(II) chloride to a DiI-supported membrane quenches the fluorescence in the top leaflet of the supported membrane; FLIC measurements of this system precisely locate the DiI to the bottom leaflet. These experiments confirm the accuracy of the model and parameters used to determine the water layer thickness, demonstrate the ability to differentiate between fluorescent objects whose average position differs by approximately 1.9 nm, and provide a widely applicable method to test the resolution of other high-z-resolution fluorescent microscopies. FLIC measurements of Alexa-labeled double-stranded oligonucleotides tethered to a supported membrane indicate that the DNA double helix is oriented perpendicular to the surface. Complications that arise from uncertainly in the orientation of the fluorophore are discussed. Several improvements in FLIC methodology are described. These stringent tests of the resolution of FLIC and the ability to unambiguously determine fluorescent lipid distribution provide structural insight on assemblies at membrane interfaces and permit the detection of even subtle changes at such interfaces. PMID- 15896040 TI - Self-assembly of trimesic acid at the liquid-solid interface-a study of solvent induced polymorphism. AB - A scanning tunneling microscope operated under ambient conditions was utilized to study the self-assembly of trimesic acid (TMA) at the liquid-solid interface. On a graphite substrate, two different open, loosely packed, two-dimensional hydrogen-bond networks were found. Both structures exhibit a periodic arrangement of approximately 1.0 nm wide cavities, which can be used for the co-adsorption of another species (guest) within the cells of this host system. These two polymorphs ("chickenwire" and "flower" structures) differ in their molecular packing density and hydrogen-bonding schemes. Using a homologous series of alkanoic acids as solvents, ranging from butyric to nonanoic, selective self assembly of either the "flower" or "chickenwire" forms was achieved on a graphite surface. Solubility of TMA in these acid solvents was found to decrease with increasing chain length, and the longer-chain solvents favored formation of the chickenwire polymorph structure on the surface. PMID- 15896041 TI - Surface ordering of a perfluorinated, self-assembled, dendrimer on a water subphase. AB - We have investigated the surface ordering of a synthetic, asymmetric, fan-shaped dendrimer containing a carboxyl core and perfluorinated tails which was obtained by the esterification of the intermediary. X-ray diffraction patterns and transmission electron microscopy (TEM) images show the molecules self-assemble into a hexagonal, cylindrical mesophase. Surface pressure-area isotherms and Brewster angle microscopy measurements show the molecule forms a stable monolayer at the air-water interface with a single phase transition. As a condensed monolayer, the perfluorinated tails are well-packed with hexagonal symmetry with (10) spacing of approximately 0.5 nm from molecular-scale atomic force microscopy (AFM) images. Such dense molecular-scale packing has not been observed in other dendritic molecules thus far. Compared to the case of conventional dendritic molecules with alkyl tails, these molecules occupy a much smaller molecular area due to the strong microphase separation between the carboxylic core and perfluorinated tails at the air-water interface. After monolayer collapse, the irregular islands with terrace morphology are observed in contrast with conventional alkyl-terminated self-assembled dendritic molecules where irregular islands do not appear. The interfacial and internal structure of every terrace shows planar columnar morphology from AFM and TEM imaging. From these results, we discuss the stability of perfluorinated, self-assembled dendrimers on water, as well as how to generate planar morphology on a hydrophilic surface. PMID- 15896042 TI - Direct and indirect monitoring of peptide-silica interactions using time-resolved fluorescence anisotropy. AB - The present work extends the application of time-resolved fluorescence anisotropy (TRFA) of a cationic probe rhodamine 6G (R6G) in aqueous Ludox to in situ monitoring of peptide adsorption onto the silica particles. Steady-state anisotropy and TRFA of R6G in Ludox sols were measured to characterize the extent of the ionic binding of the probe to silica particles in the presence of varying levels of tripeptides of varying charge, including Lys-Trp-Lys (KWK), N acetylated Lys-Trp-Lys (Ac-KWK), Glu-Trp-Glu (EWE), and N-acetylated Glu-Trp-Glu (Ac-EWE). The results were compared to those obtained by direct observation of peptide adsorption using the steady-state anisotropy of the intrinsic tryptophan residue. Ionic binding of the peptides to Ludox particles produced an increase in the steady-state Trp anisotropy that was dependent on the number of cationic groups present, but the limiting anisotropy values were relatively low, indicating significant rotational freedom of the indole residue in the adsorbed peptides. On the other hand, R6G showed significant decreases in anisotropy in the presence of cationic peptides, consistent with the cationic peptides blocking the adsorption of the dye to the silica surface. Thus, R6G is able to indirectly report on the binding of peptides to Ludox particles. It was noteworthy that, while there were similar trends in the data obtained from steady-state anisotropy and TRFA studies of R6G, the use of steady-state anisotropy to assess binding of peptides overestimated the degree of peptide adsorption relative to the value obtained by TRFA. The study shows that the competitive binding method can be used to assess the binding of various biologically relevant compounds onto silica surfaces and demonstrates the potential of TRFA for probing peptide-silica and protein-silica interactions. PMID- 15896043 TI - Adsorption of organic matter at mineral/water interfaces. IV. Adsorption of humic substances at boehmite/water interfaces and impact on boehmite dissolution. AB - The adsorption of Suwannee River fulvic acid (SRFA) and Pahokee peat humic acid (PPHA) at the boehmite (gamma-AlOOH)/water interface and the impact of SRFA on boehmite dissolution have been examined over a wide range of solution pH conditions (pH 2-12), SRFA surface coverages (Gamma(SRFA), total SRFA binding site concentration normalized by the boehmite surface area) of 0.0-5.33 micromol m(-2), and PPHA surface coverages (Gamma(PPHA), PPHA binding site concentration normalized by boehmite surface area) of 0.0-4.0 micromol m(-2), using macroscopic adsorption and in situ attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. At relatively high SRFA surface coverages (Gamma(SRFA) = 5.33 micromol m(-2)), in situ ATR-FTIR spectral features of adsorbed SRFA are very similar to those measured for SRFA in solution at approximately 1-3 pH units higher. At sub-monolayer surface coverages (Gamma(SRFA) = 1.20 and 2.20 micromol m(-2)), several new peaks and enhancements of the intensities of a number of existing peaks are observed. The latter spectral changes arise from several nonorganic extrinsic species (i.e., adsorbed carbonate and water, for alkaline solution conditions), partially protonated SRFA carboxyl functional groups (near neutral pH conditions), and small quantities of inner-spherically adsorbed SRFA carboxyl groups and/or Al(III)-SRFA complexes (for acidic conditions). The spectra of PPHA adsorbed at boehmite/water interfaces also showed changes generally consistent with our observations for SRFA sorbed on boehmite. These observations confirm that SRFA and PPHA are predominantly adsorbed at the boehmite/water interface in an outer-sphere fashion, with minor inner-sphere adsorption complexes being formed only under quite acidic conditions. They also suggest that the positively charged boehmite/water interface stabilizes SRFA and PPHA carboxyl functional groups against protonation at lower pH. Measurements of the concentration of dissolved Al(III) ions in the absence and presence of SRFA showed that the boehmite dissolution process is clearly inhibited by the adsorption of SRFA, which is consistent with previous observations that outer spherically adsorbed organic anions inhibit Al-(oxyhydr)oxide dissolution. PMID- 15896044 TI - Kinetic control of the photochemical reactivity of hydrogen-terminated silicon with bifunctional molecules. AB - Carboxy-terminated crystalline silicon surfaces are of fundamental importance for biochip fabrication because of their reactivity toward biological macromolecules. To explore the feasibility of direct attachment of bifunctional molecules (e.g., omega-alkenoic acids) to hydrogen-terminated silicon crystal (H-Si) via Si-C linkages, we have investigated the photoreactivities of the alkene (-CH=CH(2)) and carboxy (-COOH) terminal groups of 1-dodecene, undecanoic acid, and undecylenic acid toward H-Si. The alkene terminus was found to react substantially faster than the carboxy terminus under UV irradiation (at 350 nm). By controlling the reaction time, high-quality carboxy-terminated monolayers, comparable to those formed by ester hydrolysis, can be obtained from a direct, one-step photochemical reaction between H-Si and undecylenic acid. PMID- 15896045 TI - Atomic force microscopy studies of mesoscopic membranous bubbles on monolayers derived from SiCl3-terminated carbosilane dendrons on mica. AB - Monolayers of dendrimers were prepared on mica by spin-coating of the second generation carbosilane dendrons with 9 SiCl(3) periphery groups. AFM images of the films showed the presence of soft yet robust, dome-shaped features with a base diameter of 100-2000 nm. The apparent height of the features, ranging from 10 to 200 nm, rapidly reduced under increasing compression force, eventually to the same value ( approximately 2.5 nm) corresponding to a bilayer of the flattened dendrons. The change in shape of the features in response to the compression force from the AFM tip was fully reversible, indicating that the features were robust. The contrast of the features in the tapping mode AFM (TMAFM) phase images flipped at a setpoint ratio of approximately 0.55. In contrast to the reported amplitude vs displacement (A/z) curves for compliant materials, A/z curves of the features showed that the reduction of amplitude was larger than the tip displacement as if the cantilever tip were repelled by the soft features. This result cautions the use of amplitude/phase vs displacement (APD) curves for interpreting TMAFM images and for optimizing conditions for TMAFM imaging of very soft and "sticky" surfaces. On the basis of the AFM studies, we believe that the dome-shaped features are membranous air bubbles. The membranes of the bubbles were probably composed of a bilayer of the dendron molecules bound through the peripheral silanol groups. The bilayer could be formed by self-assembly of the molecules on top of the air bubbles entrapped at the monolayer/solution interface during spin-coating. PMID- 15896046 TI - Alkyl chain conformation and the electronic structure of octyl heavy chalcogenolate monolayers adsorbed on Au(111). AB - Adsorption states of dioctyl dichalcogenides (dioctyl disulfide, dioctyl diselenide, and dioctyl ditelluride) arranged on Au(111) have been studied by X ray photoelectron spectroscopy (XPS), infrared-visible sum-frequency generation (SFG), and ultraviolet photoelectron spectroscopy (UPS). XPS measurements suggest that dioctyl dichalcogenides dissociatively adsorbed on Au(111) surfaces to form the corresponding monolayers having chalcogen-gold covalent bonds. The elemental compositions of octanechalcogenolates on Au(111) indicate that the saturation coverages of the octyl heavy chalcogenolate (OcSe, OcTe) monolayers are lower than that of the octanethiolate (OcS) self-assembled monolayers (SAMs). The SFG observations of the CH(2) vibrational bands for the heavy chalcogenolate monolayers strongly suggest that a discernible amount of gauche conformation exists in the monolayers, while OcS SAMs adopt highly ordered all-trans conformation. The intensity ratio of the symmetric and asymmetric CH(3) stretching vibration modes measured by SFG shows that the average tilt angle of the methyl group of the OcSe monolayers is greater than that of the OcS SAMs. The larger tilt angle of the methyl group and the existence of a discernible amount of gauche conformation in the OcSe monolayers are due to the lower surface coverage of the OcSe monolayers compared with the OcS SAMs. The smaller polarization dependence in the angle-resolved UPS (ARUPS) spectra of the OcSe monolayers than that of the OcS SAMs is caused by the more disordered structures of the alkyl chain in the former. XPS, SFG, and ARUPS measurements indicate a similar tendency for the OcTe monolayers. The density of states (DOS) observed by UPS at around 1.3 eV for OcS adsorbed on Au(111) is considered to be the antibonding state of the Au-sulfur bond. Similar DOS is also observed by UPS at around 1.0 eV for the OcSe monolayers and at approximately 1.6 eV for the OcTe monolayers on Au(111). PMID- 15896047 TI - Self-assembled diisocyanide monolayer films on gold and palladium. AB - Self-assembled monolayers (SAMs) of the aromatic diisocyanides, 1,4 phenylenediisocyanide, 2,3,5,6-tetramethyl-1,4-phenylenediisocyanide, 4,4' biphenyldiisocyanide, 3,3',5,5'-tetramethyl-4,4'-biphenyldiisocyanide, and 4,4' ' p-terphenyldiisocyanide, were prepared on gold and palladium surfaces. The SAMs were characterized by ellipsometry, polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), and grazing-angle attenuated total reflectance infrared spectroscopy (GATR). Based on the position of the metal coordinated isocyanide stretching band, the SAMs on gold were found to bind in the terminal (eta(1)) geometry, while the SAMs on palladium prefer a different geometry which is possibly a triply bridging (mu(3)-eta(1)) geometry. A side reaction of the unbound isocyanide in the SAM was identified as oxidation to an isocyanate group. PMID- 15896048 TI - Uniformly carbon-covered alumina and its surface characteristics. AB - Uniformly carbon-covered alumina (CCA) was prepared via the carbonization of sucrose highly dispersed on the alumina surface. The CCA samples were characterized by XRD, XPS, DTA-TG, UV Raman, nitrogen adsorption experiments at 77 K, and rhodamine B (RB) adsorption in aqueous media. UV Raman spectra indicated that the carbon species formed were probably conjugated olefinic or polycyclic aromatic hydrocarbons, which can be considered molecular subunits of a graphitic plane. The N(2) adsorption isotherms, pore size distributions, and XPS results indicated that carbon was uniformly dispersed on the alumina surface in the as-prepared CCA. The carbon coverage and number of carbon layers in CCA could be controlled by the tuning of the sucrose content in the precursor and impregnation times. RB adsorption isotherms suggested that the monolayer adsorption capacity of RB on alumina increased drastically for the sample with uniformly dispersed carbon. The as-prepared CCA possessed the texture of alumina and the surface properties of carbon or both carbon and alumina depending on the carbon coverage. PMID- 15896049 TI - Pulsed-flow microcalorimetric study of the template-monolayer region of nonionic surfactants adsorbed at the graphite/water interface. AB - The formation of half-cylindrical surfactant aggregates at the graphite/aqueous solution interface is templated by an ordered monolayer of molecules disposed parallel to the graphite basal plane. Beyond a critical alkyl chain length, monolayer formation is effectively irreversible. Since enthalpic interactions in this template-monolayer region cannot be resolved with adequate accuracy by the traditional adsorption calorimetric methods, we applied a novel method, pulsed flow calorimetry, for simultaneous measurement of the material balance and the enthalpy balance in this high-affinity region. For the three nonionic surfactants studied, n-octyl beta-D-glucoside (C(8)G(1)), dimethyl-n-decylamine oxide (C(10)DAO), and n-octyl tetraethylene glycol monoether (C(8)E(4)), the adsorption was found to be strongly exothermic and effectively irreversible at low adsorbate densities, and the differential heat of adsorption markedly decreased with increasing surface coverage in this region. This deviation from the ideal adsorption behavior was attributed to intermolecular interactions within the adsorption layer rather than to surface heterogeneity of the graphite basal planes. A thermodynamic consistency test clearly demonstrated that pulsed-flow calorimetry is a unique experimental method for the study of nonreversible adsorption phenomena at solid/solution interfaces, representing an excellent tool to complement traditional methods, e.g., frontal-flow and titration adsorption calorimetry. Studies by the frontal-flow method revealed that aggregation on top of the surfactant monolayer was endothermic and reversible. PMID- 15896050 TI - Predicting adsorption isotherms of low-volatile compounds by temperature programmed desorption: iodine on carbon. AB - Equilibrium adsorption isotherms for low-volatile compounds are extremely difficult to measure. A simple technique using temperature programmed desorption (TPD) is proposed. It is demonstrated that the two parameters needed for constructing the Langmuir isotherm can be derived with data from the TPD technique alone. Thus, the Langmuir isotherms of iodine on AX-21 super-activated carbon were obtained with this technique. A series of TPD experiments for samples with different initial loadings of iodine were carried out by varying the heating rates which resulted in different peak desorption temperatures. The peak desorption temperature decreased as the initial loading was increased because of the re-adsorption effect. The Langmuir constant was derived from kinetic theory with the activation energy for desorption obtained from the experiment. The activation energy for desorption was 12.3 kcal/mol. The Langmuir constants determined by this technique were in comparable order of magnitude to the reported values for iodine on activated carbon. The saturation capacity of AX-21 for iodine could also be determined from the TPD data obtained from samples with different initial loadings. The estimated saturation capacity from the TPD experiment was 2.96 g I(2)/g AX-21, which was close to the experimentally measured saturation capacity of 3.25 g I(2)/g AX-21 for the same system. PMID- 15896051 TI - Ellipsometric characterization of ethylene oxide-butylene oxide diblock copolymer adsorption at the air-water interface. AB - Ellipsometry was used to determine the adsorbed layer thickness (d) and the surface excess (adsorbed amount, Gamma) of a nonionic diblock copolymer, E(106)B(16), of poly(ethylene oxide) (E) and poly(butylene oxide) (B) at the air water interface. The results were obtained (i) by the conventional ellipsometric evaluation procedure using the change of both ellipsometric angles Psi and Delta and (ii) by using the change of Delta only and assuming values of the layer thickness. It was demonstrated that the calculated surface excesses from the different methods were in close agreement, independent of the evaluation procedure, with a plateau adsorption of about 2.5 mg/m(2) (400 A(2)/molecule). Furthermore, the amount of E(106)B(16) adsorbed at the air-water interface was found to be almost identical to that adsorbed from aqueous solution onto a hydrophobic solid surface. In addition, the possibility to use combined measurements with H(2)O or D(2)O as substrates to calculate values of d and Gamma was investigated and discussed. We also briefly discuss within which limits the Gibbs equation can be used to determine the surface excess of polydisperse block copolymers. PMID- 15896052 TI - Dynamics of ultrathin films in the glassy state. AB - We report hole growth experiments in free-standing polystyrene (PS) films at temperatures up to 10 degrees C below the bulk glass transition. The data show an unexpected result: the growth rate of nucleated holes increases with increasing molecular weight, up to a limiting value beyond which the rate is approximately constant. Film thicknesses of 45, 80, and 100 nm were studied, using PS molecular weights ranging from 65K to 11.4 Mg/mol. Hole diameters grew linearly with time, and no growing rims were observed to form around the developing holes. Possible explanations in terms of elasticity, yield, and influence of sample preparation and confinement effects are discussed. PMID- 15896053 TI - Tuning the surface hydrophobicity of polymer/nanoparticle composite films in the Wenzel regime by composition. AB - Surface hydrophobicity of composite films containing polymer and nanoparticles has been studied as a function of composition. We show that the hydrophobicity can be tuned by adjusting the amount of particles in the two-component system. A sharp transition from a polymer-rich surface to a nanoparticles-rich surface was observed with increasing mass fraction of particles in spin-coated thin films. Water drops on the films did not slide down even at tilt angles of 90 degrees . Contact angle hysteresis increased with the mass fraction of particles indicating that the surface roughness increased as the surfaces remained in the Wenzel regime. Contact angle hysteresis data were quantitatively consistent with predictions of a recent theory. PMID- 15896054 TI - Controllable growth of well-defined regular multiporphyrin array nanocrystals at the water-chloroform interface. AB - On the basis of the coordination geometry of metal ions, regular cubic, clubbed, and wirelike nanocrystals of Cd(2+)-/PtCl(6)(2-)-mediated, and Hg(2+)-/Ag(+) /PtCl(4)(2-)-mediated multiporphyrin arrays have been grown at the water chloroform interface. The nanocrystal growth process was monitored by the transmission electron microscopy (TEM), which revealed (1) an intrinsic rule for coordination polymers, that is, the geometries of metal ions (as connects for the coordination polymers) dominate the frameworks of the related polymeric nanocrystals, and (2) one kind of intuitive nanocrystal growth processes at the interfaces. Both electron diffraction and X-ray diffraction patterns indicated the formation of well-defined nanocrystals. It was found that single /microcrystals were formed at first, and then they grew into polycrystals. The nanocrystal layer was transferred onto Si and quartz substrate surfaces by the Langmuir-Blodgett method, with its composition analyzed by X-ray photoelectron spectroscopy as well as the arrangement of porphyrin macrocycles in the nanocrystals by UV-vis absorption spectroscopy. PMID- 15896055 TI - Alkane/Alcohol mixed monolayers at the solid/liquid interface. AB - In this work, we present the behavior of solid monolayers of binary mixtures of alkanes and alcohols adsorbed on the surface of graphite from their liquid mixtures. We demonstrate that solid monolayers form for all the combinations investigated here. Differential scanning calorimetry (DSC) is used to identify the surface phase behavior of these mixtures, and elastic neutron incoherent scattering has been used to determine the composition of the mixed monolayers inferred by the calorimetry. The mixing behavior of the alcohol/alkane monolayer mixtures is compared quantitatively with alkane/alkane and alcohol/alcohol mixtures using a regular solution approach to model the incomplete mixing in the solid monolayer with preferential adsorption determining the surface composition. This analysis indicates the preferential adsorption of alcohols over alkanes of comparable alkyl chain length and even preferential adsorption of shorter alcohols over longer alkanes, which contrasts strongly with mixtures of alkane/alkane and alcohol/alcohol of different alkyl chain lengths where the longer homologue is always found to preferentially adsorb over the shorter. The alcohol/alkane mixtures are all found to phase separate to a significant extent in the adsorbed layer mixtures even when molecules are of a similar size. Again, this contrasts strongly with alkane/alkane and alcohol/alcohol mixtures where, although phase separation is found for molecules of significantly different size, good mixing is found for similar size species. PMID- 15896056 TI - Adsorption of C7 hydrocarbons on biporous SBA-15 mesoporous silica. AB - In our recent studies (Vinh-Thang, H.; Huang, Q.; Eic, M.; Trong-On, D.; Kaliaguine, S. Langmuir 2005, 21, 2051-2057; Vinh-Thang, H.; Huang, Q.; Eic, M.; Trong-On, D.; Kaliaguine, S. Stud. Surf. Sci. Catal. 2005, in press), a series of synthesized SBA-15 materials were characterized using nitrogen adsorption/desorption isotherms at 77 K and SEM images. In the present paper, four of them (MMS-1-RT, MMS-1-60, MMS-1-80, and MMS-5-80) were further investigated with regard to their equilibrium characteristics using n-heptane and toluene as sorbates by the standard gravimetric technique. SBA-15 materials proved to have a broad pore size distribution within the micropore/small-mesopore range in the walls of their main mesoporous channels. The adsorption capacities for toluene were found to be higher than for n-heptane. The isosteric heats of adsorption, estimated by the Clausius-Clapeyron equation, are also higher for toluene compared to n-heptane. They were found to depend on framework microporosity of the relevant SBA-15 samples. The isosteric heats of adsorption for all sorbates decrease with increased loading and approach the heats of evaporation of the respective sorbate. The adsorption capacities of SBA-15 samples are significantly higher than those of silicalite, i.e., the MFI zeolite silica analogue. In contrast to that, the isosteric heats of adsorption in the mesopore channels of SBA-15 were found to be much smaller. This result also suggests that SBA-15 can potentially be a good candidate for separation of C(7) hydrocarbons. PMID- 15896057 TI - Phenylboronic acid monolayer-modified electrodes sensitive to sugars. AB - The surface of a gold (Au) electrode was modified with 4-mercaptophenylboronic acid (MPBA) and dithiobis(4-butyrylamino-m-phenylboronic acid) (DTBA-PBA) to prepare sugar-sensitive electrodes. MPBA and DTBA-PBA formed well-packed monomolecular layers on the Au electrode through a sulfur-Au bond. The MPBA- and DTBA-PBA-modified electrodes exhibited a nearly reversible cyclic voltammogram (CV) for the Fe(CN)(6)(3-)(/4-) ion in acidic solution, while the CVs were significantly attenuated in alkaline media as a result of addition of OH(-) ion to the boron atom to generate the negatively charged surface. In other words, the negatively charged monolayers blocked the surface of the Au electrode from the access of the Fe(CN)(6)(3-)(/4-) ion. The pK(a) values of the addition equilibrium of the OH(-) ion to the MPBA and DTBA-PBA monolayers were estimated to be 9.2 +/- 0.1 and 8.0 +/- 0.2, respectively, on the basis of the pH-dependent peak current (i(p)) in the CV of the Fe(CN)(6)(3-)(/4-) ion. On the other hand, in the presence of sugars, the addition of the OH(-) ion was accelerated by forming the phenylboronate esters of sugars on the surface of the monolayers. The pK(a) values for the MPBA and DTBA-PBA monolayers were 8.3 +/- 0.1 and 7.2 +/- 0.1, respectively, in the presence of 50 mM D-fructose. The MPBA- and DTBA-PBA modified electrodes can be used for determining sugars on the basis of the change in i(p) of the Fe(CN)(6)(3-)(/4-) ion in the presence of sugars. The calibration curves useful for determining 1-100 mM D-glucose, D-mannose, and D-fructose were obtained. PMID- 15896058 TI - Electrokinetic characterization of poly(acrylic acid) and poly(ethylene oxide) brushes in aqueous electrolyte solutions. AB - Surfaces carrying hydrophilic polymer brushes were prepared from poly(styrene) poly(acrylic acid) and poly(styrene)-poly(ethylene oxide) diblock copolymers, respectively, using a Langmuir-Blodgett technique and employing poly(styrene) coated planar glass as substrates. The electrical properties of these surfaces in aqueous electrolyte were analyzed as a function of pH and KCl concentration using streaming potential/streaming current measurements. From these data, both the zeta potential and the surface conductivity could be obtained. The poly(acrylic acid) brushes are charged due to the dissociation of carboxylic acid groups and give theoretical surface potentials of -160 mV at full dissociation in 10(-)(3) M solutions. The surface conductivity of these brushes is enormous under these conditions, accounting for more than 93% of the total measured surface conductivity. However, the mobility of the ions within the brush was estimated from the density of the carboxylic acid groups and the surface conductivity data to be only about 14% of that of free ions. The poly(ethylene oxide) (PEO) brushes effectively screen the charge of the underlying substrate, giving a very low zeta potential except when the ionic strength is very low. From the data, a hydrodynamic layer thickness of the PEO brushes could be estimated which is in good agreement with independent experiments (neutron reflectivity) and theoretical estimates. The surface conductivity in this system was slightly lower than that of the polystyren substrate. This also indicates that no significant amount of preferentially, i.e., nonelectrostatically attracted, ions taken up in the brush. PMID- 15896059 TI - Electrochemistry of surface-grafted stimulus-responsive monolayers of poly(ferrocenyldimethylsilane) on gold. AB - Poly(ferrocenyldimethylsilane)s with various degrees of polymerization and featuring a thiol end group were chemically end-grafted onto gold substrates by self-assembly, forming redox-active monolayers. The monolayers were characterized by contact angle measurements, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. Layer thickness values were determined by surface plasmon resonance spectroscopy and ellipsometry. The electrochemical properties of these films in aqueous NaClO(4) were studied using cyclic voltammetry (CV), differential pulse voltammetry, chronoamperometry, and chronocoulometry. Cyclic voltammograms showed two reversible redox peaks, indicating a stepwise oxidation of the electroactive sites. The first oxidation step showed reversible behavior at low scan rates and quasi-reversible behavior at higher scan rates. Peak currents (i(p)) plotted against the square root of scan rates (v(1/2)) for the first oxidation peak and for the corresponding reduction peak exhibited a linear dependence, indicating that the oxidation process in the first step is controlled by the diffusion of counterions into the polymer film. For the second oxidation peak and the corresponding reduction peak, i(p) varied linearly with v. This redox behavior is characteristic of surface-immobilized electroactive layers. The higher reversibility of the second oxidation and reduction waves in the CV experiments was explained from the solvation of the surface-grafted poly(ferrocenylsilane) (PFS) chains, which depends on the degree of oxidation. Oxidized PFS films are swollen in the aqueous electrolyte solutions, leading to a higher segmental mobility of the polymer chains and a much increased counterion mobility within the film. Kinetic parameters for the redox processes were obtained from chronocoulometry experiments. PMID- 15896060 TI - Electrocrystallization of rhodium clusters on thiolate-covered polycrystalline gold. AB - This study reports on the electrochemical deposition of rhodium metal clusters on a polycrystalline gold electrode, modified with a monolayer of dodecanethiol through self-assembly from solution. The deposition process was investigated using cyclic voltammetry, chronoamperometry, and electrochemical quartz crystal microbalance. It is shown that the presence of the thiol monolayer drastically alters the nucleation and growth mechanism compared with the mechanism on the bare gold electrode. The small uncovered gold domains, located at the imperfections in the thiolate monolayer which are induced by the gold nanoroughness, act as nucleation sites for small rhodium clusters. At longer times, these clusters can outgrow the organic monolayer. The resulting surface morphology was characterized by scanning electron microscopy. Rhodium electrocrystallization on the bare gold substrate resulted in an ensemble of a very large amount of very small clusters that are difficult to distinguish from the gold roughness. In contrast, in the presence of a self-assembled monolayer (SAM) of dodecanethiol covalently attached to the gold electrode, the resulting deposit consisted of an ensemble of hemispherical particles. The size distribution of the rhodium particles obtained by using double step chronoamperometry was compared to the ones obtained with cyclic voltammetry and "classical" chronoamperometry. It is shown by X-ray photoelectron spectroscopy that the SAM is still present after rhodium deposition on the thiolate-covered gold substrate. Because the rhodium clusters are directly attached to the gold substrate and can thus easily be electrified, the resulting interface could be used as a composite electrode consisting of a random array of gold supported rhodium nano/microparticles separated from each other by an organic phase. On the other hand, it is shown that the SAM is easily removed by electrochemical oxidation without dissolving the rhodium clusters and, thus, leaving a different array of rhodium clusters on the gold surface compared with the topography obtained in the absence of the SAM. From this point of view, substrate modification with such "removable" organic monolayers was found to be an interesting tool to tune the nano- or microtopography of electrochemically deposited rhodium. PMID- 15896061 TI - Evidence of impurities in thiolated single-stranded DNA oligomers and their effect on DNA self-assembly on gold. AB - The diversity of techniques used in the synthesis, treatment, and purification of the single-stranded DNA oligomers containing a thiol anchor group (SH-ssDNA) has led to a significant variation in the purity of commercially available SH-ssDNA. In this work, we use X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to study how the impurities present in commercially synthesized SH-ssDNA oligomers affected the structure of the resulting DNA films on Au. XPS results indicate that two of the purchased SH ssDNA oligomers contain excess carbon and sulfur. The molecular fragmentation patterns obtained with ToF-SIMS were used to determine the identity of several contaminants in the DNA films, including poly(dimethylsiloxane) (PDMS), lipid molecules, and sulfur-containing molecules. In particular, the ToF-SIMS results determined that the excess sulfur detected by XPS was due to the presence of dithiothreitol, a reductant often used to cleave disulfide precursors. Furthermore, we found that the SH-ssDNA self-assembly process is affected by the presence of these contaminants. When relatively pure SH-ssDNA is used to prepare the DNA films, the P, N, O, and C atomic percentages were observed by XPS to increase over a 24-h time period. In contrast, surfaces prepared using SH-ssDNA containing higher levels of contaminants did not follow this trend. XPS result indicates that, after the initial SH-ssDNA adsorption, the remaining material incorporated into these films was due to contamination. PMID- 15896062 TI - Biophysical characterization of complexation of DNA with block copolymers of poly(2-dimethylaminoethyl) methacrylate, poly(ethylene oxide), and poly(propylene oxide). AB - The interactions of DNA (salmon testes) with two new cationic block copolymers made of poly(2-dimethylaminoethyl) methacrylate and poly(ethylene oxide), PEO pDMAEMA, or poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide), L92 pDMAEMA, were studied with the aim to understand their different in vitro transfection efficiencies when used as nonviral delivery vectors. PEO-pDMAEMA does not show surface activity while L92-pDMAEMA is as surface active as its parent Pluronic L92. Surface tension, titration microcalorimetry, ethidium bromide displacement, and zeta-potential measurements were carried out in phosphate buffers at pH 5 and 7. The association of L92-pDMAEMA with DNA was strongly exothermic at both pHs; the critical aggregation concentration (CAC) corresponded to a N/P ratio of 0.3, the maximum energy evolved was reached for N/P ratios of 0.82 and 1.27 at pH 5 and pH 7, respectively, and the saturation occurred for N/P ratios close to 2. The presence of L92 in the structure of this new block copolymer apparently did not modify the thermodynamic parameters of the interaction with DNA. In contrast, the interaction with PEO-pDMAEMA was significantly less exothermic, and CAC and saturation occurred for N/Ps equal to 0.43 and 1.37, respectively. The strong affinity of L92-pDMAEMA for DNA was reflected in its capacity to displace ethidium bromide and in the jump in the values of the zeta potential when N/P is near 1. Above the N/P ratio at which electroneutral polyplexes are formed, only at pH 5 an excess of L92-pDMAEMA is incorporated in the complexes, resulting in positively charged complexes. The profile of the zeta-potential values obtained for mixtures of L92-pDMAEMA with Pluronic P123 showed a shift to a lower N/P ratio, owing to an easier interaction of L92-pDMAEMA molecules with DNA in the presence of P123. Additionally, a visual inspection of the systems indicates that P123 contributes to stabilize/solubilize the DNA/cationic polymer aggregates, by avoiding the typical phase separation near the charge neutralization point. The information obtained can be particularly useful to optimize the conditions to form efficient polyplexes for gene delivery systems. PMID- 15896063 TI - Measurement of the anchorage force between GPI-anchored alkaline phosphatase and supported membranes by AFM force spectroscopy. AB - Mammalian alkaline phosphatases (AP) are glycosylphosphatidylinositol (GPI) anchored proteins that are localized on the outer layer of the plasma membrane. The GPI anchors are covalently attached to the C-termini of proteins and consist of a glycan chain bonded to phosphatidylinositol with two acyl chains anchored into the membrane bilayer. Force spectroscopy, based on atomic force microscope (AFM) technology, was used to determine the adhesion of alkaline phosphatase in the absence and presence of anchors. The GPI anchors increase markedly the adhesion frequency (i.e., the protein affinity for the membrane). An adhesion force of 350 +/- 200 pN is measured between GPI-anchored AP (AP(GPI)) and supported phospholipid bilayers of dipalmitoylphosphatidylcholine (DPPC) presenting structural defects (holes). In the absence of defects, the adhesion force (103 +/- 17 pN) and the adhesion frequency are reduced. These results indicate that AP(GPI) poorly spontaneously insert into membranes in vivo and open new perspectives for the characterization of the interactions between GPI proteins and membranes. PMID- 15896064 TI - Conformation of microcontact-printed proteins by atomic force microscopy molecular sizing. AB - We investigated the structural changes occurring in proteins patterned via microcontact printing. This was done by molecular sizing using atomic force microscopy to observe the structure of printed individual metalloprotein molecules in the unlabeled and untreated states. We observed that the size of the printed proteins were more than 2-fold smaller than the native shape, which indicates that some deformations take place upon the contact-assisted adsorption on silanized silicon dioxide. This can be attributed to simultaneously occurring effects, and particularly to the sandwiching between surfaces of very different hydrophilic/hydrophobic properties during contact lithography. PMID- 15896065 TI - Aqueous phase behavior and dispersed nanoparticles of diglycerol monooleate/glycerol dioleate mixtures. AB - The first part of this study concerns the aqueous phase behavior of mixtures of diglycerol monooleate (DGMO) and glycerol dioleate (GDO) examined by X-ray diffraction (XRD). The ternary phase diagram displays a multitude of liquid crystalline phases (polymorphism). With increasing GDO content the following phase sequence was observed: lamellar (L(alpha)); two reversed bicontinuous cubic phases (Q(230) and Q(224)); reversed hexagonal (H(II)); the reversed micellar (L(2)) phase. The second part deals with the preparation and characterization of aqueous dispersions of the reversed hexagonal phase in the presence of the nonionic triblock copolymer Pluronic F127. Submicrometer-sized monocrystalline H(II) phase particles were obtained, as evidenced by cryo-transmission electron microscopy (cryo-TEM), laser diffraction, and XRD, by use of a simple and reproducible preparation method including a heat-treatment step. Moreover, the particle size distributions of the H(II) phase nanoparticle dispersions were narrow as determined by laser diffraction measurements. Using XRD, we show that the polymeric stabilizer is depleted from the core of the hexagonal particles and preferentially located at the surface. It is concluded that the preferential distribution of stabilizing agents at particle surfaces is a prerequisite for the formation of structurally well-defined and kinetically stable H(II) phase particles (Hexosome). PMID- 15896066 TI - Chloride ion effects on synthesis and directed assembly of copper nanoparticles in liquid and compressed alkane microemulsions. AB - Microemulsions are effective media for solution-based synthesis of metallic nanoparticles where surfactants and other ionic species influence the directed assembly of the nanomaterials with specific sizes, geometries, and compositions. This study demonstrates the effects of chloride ion on the synthesis of copper nanoparticles within the sodium bis(2-ethylhexyl)sulfosuccinate (AOT) reverse micelle system utilizing both liquid isooctane and compressed propane as the bulk solvent. Copper nanoparticle synthesis can be achieved in the presence of HCl in the micelle core, taking advantage of the buffering action of the AOT surfactant. The concentration of chloride ions influence the particle growth rate and dispersion in liquid isooctane. The presence of chloride ions during particle synthesis in compressed propane has a significant effect on the geometry and structure of the copper nanomaterials produced. Chloride ion addition to the compressed propane/Cu(AOT)(2)-AOT/water reverse micelle system at 20 degrees C and 310 bar results in the formation of diamond-shaped copper nanoparticle assemblies. The copper nanoparticle assemblies exhibit unique structure and retain this structure through repeated solvent processing steps, allowing separation and recovery of the assembled diamond-shaped copper nanoparticle structures. PMID- 15896067 TI - Interaction of sodium dodecyl sulfate with methacrylate-PEG comb copolymers. AB - A series of sodium methacrylate and poly(ethylene glycol) (PEG) comb copolymers (MAA/PEG) with approximate PEG chain lengths of 7, 11, and 22 ethylene oxide units were synthesized by free radical polymerization. Their weight-average molecular mass was found to be approximately 66 000. A commercial sample of a PEG comb polymer with an acrylic backbone was also used in the studies (Sokalan HP 80). The interaction of the MAA/PEG comb polymers and pure sodium methacrylate (SPMA) with sodium dodecyl sulfate (SDS) was studied by ESR spectroscopy using 5 doxyl stearic acid (5-DSA) spin probe and by conductivity measurements. Surfactant aggregation in water occurred at SDS concentrations lower than the surfactant critical micelle concentration (cmc) and depended on the polymer concentration. The observations have been attributed to changes in the effective ionic strength of the systems due to the polymer itself, and it has been concluded that there is no interaction between the MAA/PEG comb copolymers or SPMA and SDS. This has been confirmed by the fact that the decrease in surfactant aggregation concentration is similar in magnitude to the decrease observed on adding NaCl when counterion ion condensation effects are taken into account. It is apparent that the electrostatic repulsions between the surfactant molecules and the methacrylate backbone of the MAA/PEG comb copolymers inhibit association of SDS with the PEG side chains. PMID- 15896068 TI - Selective dissolution of the silver component in colloidal Au and Ag multilayers: a facile way to prepare nanoporous gold film materials. AB - Colloidal Au/Ag multilayer films were prepared by alternate assembly of Au nanoparticles with a size of 5 +/- 1.2 nm and Ag nanoparticles with a size of 10 +/- 2.4 nm by using 1,5-pentanedithiol as cross-linker. Nanoporous gold films with a ligament size of 26.7 +/- 4.6 nm were then prepared by selective dissolution of sacrificial templates of silver particles in colloidal Au/Ag multilayers. The complete dissolution of Ag particles in colloidal Au/Ag multilayers in a mixture solution of 3.0 mM HAuCl(4) and 3 M NaCl took place at room temperature without damage of the colloidal Au film. This method to prepare nanoporous gold films was further extended to the preparation of nanoporous gold nanotubes by depositing colloidal Au/Ag film on the inner wall of anodic aluminum oxides (AAO) followed by dissolution of colloidal Ag and removal of AAO templates. PMID- 15896069 TI - Synthesis of hydroxyapatite crystals using amino acid-capped gold nanoparticles as a scaffold. AB - Inorganic composites are of special interest for biomedical applications such as in dental and bone implants wherein the ability to modulate the morphology and size of the inorganic crystals is important. One interesting possibility to control the size of inorganic crystals is to grow them on nanoparticles. We report here the use of surface-modified gold nanoparticles as templates for the growth of hydroxyapatite crystals. Crystal growth is promoted by a monolayer of aspartic acid bound to the surface of the gold nanoparticles; the carboxylate ions in aspartic acid are excellent binging sites for Ca(2+) ions. Isothermal titration calorimetry studies of Ca(2+) ion binding with aspartic acid-capped gold nanoparticles indicates that the process is entropically driven and that screening of the negative charge by the metal ions leads to their aggregation. The aggregates of gold nanoparticles are believed to be responsible for assembly of the platelike hydroxyapatite crystals into quasi-spherical superstructures. Control experiments using uncapped gold nanoparticles and pure aspartic acid indicate that the amino acid bound to the nanogold surface plays a key role in inducing and directing hydroxyapatite crystal growth. PMID- 15896070 TI - Understanding the dynamics of signal transduction for adsorption of gases and vapors on carbon nanotube sensors. AB - Adsorption dynamics and their influence on signal transduction for carbon nanotube-based chemical sensors are explored using continuum site balance equations and a mass action model. These sensors are shown to possess both reversible and irreversible binding sites that can be modeled independently. For the case of irreversible adsorption, it is shown that the characteristic response time scales inversely with analyte concentration. It is inappropriate to report a detection limit for this type of sensor since any nonzero analyte concentration can be detected in theory but at a cost of increasing transduction time with decreasing concentration. The response curve should examine the initial rate of signal change as a function of analyte concentration. Conversely, a reversible sensor has a predefined detection limit, independent of the detector geometry with a characteristic time scaling that becomes constant in the zero analyte concentration limit. A simple analytical test is presented to distinguish between these two mechanisms from the transient response of a nanotube sensor array. Two systems appearing in the literature are shown to have an irreversible component, and regressed surface rate constants for this component are similar across different sensor geometries and analytes. PMID- 15896071 TI - Formation and structure of self-assembled silica nanoparticles in basic solutions of organic and inorganic cations. AB - The phase behavior of silica solutions containing organic and inorganic cations was studied at room temperature using conductivity, pH, and small-angle scattering experiments. A critical aggregation concentration (cac) was observed at approximately 1:1 ratio of SiO(2)/OH(-) for all cation solutions from conductivity and pH studies. From this cac, a phase diagram of the system was developed with three distinct phase regions in pseudoequilibrium: a monomer/oligomer region (I), a monomer/oligomer/nanoparticle region (II), and a gel region (III). Small-angle X-ray and neutron scattering (SAXS and SANS) on solutions of region II formed with tetrapropylammonium hydroxide (TPAOH) revealed that the nanoparticles have a core-shell structure. Structure analysis of the SAXS and SANS data was best fit by a core-shell oblate ellipsoid model. A polydisperse set of core-shell spheres also fit the data well although with lower agreement factors. Similar nanoparticle morphologies were found in solutions of TMAOH, CsOH, and NaOH. PMID- 15896072 TI - Luminescent core-shell photonic crystals from poly(phenylene ethynylene) coated silica spheres. AB - We describe the preparation and characterization of a photonic crystal filled with a luminescent conjugated polyelectrolyte, sulfonated poly(phenylene ethynylene). The conjugated polymer was coated onto the nanospheres by the layer by-layer method and assembled directly into a fluorescent opal structure avoiding the defects associated with post-filling schemes. These structures exhibit strong angle-dependent luminescent properties. By using multiple layers, we further demonstrate control over the emissive bands of the opal. PMID- 15896073 TI - High-density arrays of titania nanoparticles using monolayer micellar films of diblock copolymers as templates. AB - Highly dense arrays of titania nanoparticles were fabricated using surface micellar films of poly(styrene-block-2-vinylpyridine) diblock copolymers (PS-b P2VP) as reaction scaffolds. Titania could be introduced selectively within P2VP nanodomains in PS-b-P2VP films through the binary reaction between water molecules trapped in the P2VP domains and the TiCl(4) vapor precursors. Subsequent UV exposure or oxygen plasma treatment removed the organic matrix, leading to titania nanoparticle arrays on the substrate surface. The diameter of the titania domains and the interparticle distance were defined by the lateral scale present in the microphase-separated morphology of the initial PS-b-P2VP films. The typical diameter of titania nanoparticles obtained by oxygen plasma treatment was of the order of approximately 23 nm. Photoluminescence (PL) properties were investigated for films before and after plasma treatment. Both samples showed PL properties with major physical origin due to self-trapped excitons, indicating that the local environment of the titanium atoms is similar. PMID- 15896074 TI - Semiconductor nanoparticle/polystyrene latex composite materials. AB - Cadmium sulfide and cadmium selenide/cadmium sulfide core/shell nanoparticles stabilized with poly(cysteine acrylamide) have been bound to polystyrene (PS) latexes by three methods. First, anionic 5 nm diameter CdS particles were electrostatically attached to 130 nm surfactant-free cationic PS latexes to form stable dispersions when the amount of CdS particles was less than 10% of the amount required to form a monolayer on the surface of the PS particles or when the amount of CdS particles exceeded the amount required to form a monolayer on the PS particles. Transmission electron microscopy (TEM) showed nanoparticles on the surface of the latex particles. Fluorescence spectra showed unchanged emission from the nanoparticles. Second, anionic, surfactant-free PS latexes were synthesized in the presence of CdS and CdSe/CdS nanoparticles. TEM showed monodisperse latex particles with trapped nanoparticles. Third, surfactant stabilized latexes were synthesized by copolymerization of styrene with vinylbenzyl(trimethyl)ammonium chloride electrostatically bound to the CdSe/CdS nanoparticle surface. Brownian motion of the submicroscopic composite particles in water was detected by fluorescence microscopy. PMID- 15896075 TI - From homogeneous dispersion to micelles-a molecular dynamics simulation on the compromise of the hydrophilic and hydrophobic effects of sodium dodecyl sulfate in aqueous solution. AB - The structural and functional diversity of surfactant systems has attracted simulation works in atomistic, coarse grain, and mesoscopic models (Bandyopadhyay, S.; et al. Langmuir 2000, 16, 942; Senapati, S.; et al. J. Phys. Chem. B 2003, 107, 12906; Maiti, P. K.; et al. Langmuir 2002, 18, 1908; Srinivas, G.; et al. J. Phys. Chem. B 2004, 108, 8153; Groot, R. D.; et al. J. Chem. Phys. 1999, 110, 9739; Rekvig, L.; et al. Langmuir 2003, 19, 8195). However, atomistic models have suffered from their tremendous computational cost and are, so far, not able to simulate the structural behaviors in sufficient spatio-temporal scales (Shelley, J. C.; Shelley, M. Y. Curr. Opin. Colloid Interface Sci. 2000, 5, 101). The other two approaches are not microscopic enough to describe the configurations of the surfactants that determine their behaviors (Shelley and Shelley). In this study, we propose to simplify atomistic models based on the observation that the compromise of the hydrophilic and hydrophobic effects (Li, J.; Kwauk, M. Chem. Eng. Sci. 2003, 58, 521-535) and molecular structures of surfactants are the dominant factors shaping their structures in the systems. With this simplification, we are able to simulate with moderate computing cost the whole process of micelle formation from an initially uniform dispersion of sodium dodecyl sulfate (SDS) in aqueous solution. The resulting micelle structures are different from those predicted by atomistic simulations that started with a predefined micelle configuration at the same surfactant concentrations. However, if we use their initial micelle configuration, micelle structures the same as theirs are obtained. Analyses show that our results are more realistic and that the results of the atomistic simulations suffer from artificial initial conditions. Therefore, our model may serve as a reasonable simplification of atomistic models in terms of the general structure of micelles. PMID- 15896076 TI - Initial nitridation of the Ge(100)-2 x 1 surface by ammonia. AB - The reaction of ammonia (NH(3)) on the Ge(100)-2 x 1 surface is investigated using density functional theory (DFT). We find that NH(3) adsorbs molecularly onto Ge(100)-2 x 1 via the formation of a dative bond. The calculations also show that, unlike Si(100)-2 x 1, the activation barrier for subsequent dissociation of NH(3) adsorbed on Ge(100)-2 x 1 is higher than that of reversible desorption, which indicates that NH(3) has a low reactive sticking probability on the Ge(100) 2 x 1 surface. We also predict that nitrogen insertion into the Ge-Ge dimer requires NH(3) overexposure because the activation barrier for NH(2) insertion into the Ge-Ge dimer is significantly above the entrance channel. The nitridation reaction pathway results in the N-H bridge-bonded state, which is found to be 17.4 kcal/mol more stable than the reactants. We find that the reactions of NH(3) on the Ge(100)-2 x 1 surface generally involve higher activation barriers and less stable intermediates than the analogous reactions on the Si(100)-2 x 1 surface. PMID- 15896077 TI - Optical detection of tumors in vivo by visible light tissue oximetry. AB - Endoscopy is a standard procedure for identifying tumors in patients suspected of having gastrointestinal (G.I.) cancer. The early detection of G.I. neoplasms during endoscopy is currently made by a subjective visual inspection that relies to a high degree on the experience of the examiner. This process can be difficult and unreliable, as tumor lesions may be visually indistinguishable from benign inflammatory conditions and the surrounding mucosa. In this study, we evaluated the ability of local ischemia detection using visible light spectroscopy (VLS) to differentiate neoplastic from normal tissue based on capillary tissue oxygenation during endoscopy. Real-time data were collected (i) from human subjects (N = 34) monitored at various sites during endoscopy (enteric mucosa, malignant, and abnormal tissue such as polyps) and (ii) murine animal subjects with human tumor xenografts. Tissue oximetry in human subjects during endoscopy revealed a tissue oxygenation (StO2%, mean +/- SD) of 46 +/- 22% in tumors, which was significantly lower than for normal mucosal oxygenation (72 +/- 4%; P < or = 0.0001). No difference in tissue oxygenation was observed between normal and non-tumor abnormal tissues (P = N.S.). Similarly, VLS tissue oximetry for murine tumors revealed a mean local tumor oxygenation of 45% in LNCaP, 50% in M21, and 24% in SCCVII tumors, all significantly lower than normal muscle tissue (74%, P < 0.001). These results were further substantiated by positive controls, where a rapid real-time drop in tumor oxygenation was measured during local ischemia induced by clamping or epinephrine. We conclude that VLS tissue oximetry can distinguish neoplastic tissue from normal tissue with a high specificity (though a low sensitivity), potentially aiding the endoscopic detection of gastrointestinal tumors. PMID- 15896078 TI - Optical tomography with ultrasound localization: initial clinical results and technical challenges. AB - Optical tomography with ultrasound localization utilizes co-registered ultrasound lesion structure information to guide optical imaging reconstruction. A hand-held probe consisting of a commercial ultrasound transducer and near infrared optical imaging sensors was used to simultaneously acquire ultrasound images and optical measurements. A dual-mesh scheme was used to map the ultrasound-visible lesions to finer-grid lesion regions and coarser-grid background regions for optical imaging reconstruction. As a result, optical imaging reconstruction was well conditioned for inverse mapping of lesion hemoglobin concentration and blood oxygen saturation. Initial clinical results have shown that early stage invasive cancers may be distinguished by a two-fold greater total hemoglobin concentration compared with fibroadenomas and other benign lesions. Initial results of advanced cancers have shown that the hemoglobin distribution is highly distorted and heterogeneous and the distorted distributions correlate with histological microvessel density counts and could be used to assess chemotherapy response. PMID- 15896079 TI - Dexamethasone normalizes brain tumor hemodynamics as indicated by dynamic susceptibility contrast MRI perfusion parameters. AB - The purpose of this study is to demonstrate the utility of dynamic susceptibility contrast (DSC) MRI-derived perfusion parameters to characterize the hemodynamic effects of dexamethasone in a 9L gliosarcoma tumor model. Twenty-four rats underwent intracerebral inoculation with 9L tumor cells. Fifteen were treated with a total of 3mg/kg of dexamethasone on days 10-14 post-inoculation, while the remaining 9 rats served as controls. Fourteen days post-inoculation, MRI images, sensitive to total and micro-vascular cerebral blood flow (CBF), mean transit time (MTT), and intravoxel transit time distributions (TTD)s were obtained using a simultaneous gradient-echo(GE)/spin-echo(SE) DSC-MRI method. Dexamethasone treated animals had a microvascular (SE) tumor CBF that was 45.9% higher (p = 0.0008) and a MTT that was 47.8% lower (p = 0.0005) than untreated animals. With treatment, there was a non-significant 91.3% increase in total (GE) vascular CBF (p = 0.35), and a significant decrease in MTT (49.1%, p = 0.02). The total vascular and microvascular TTDs from the treated tumors were similar to normal brain, unlike the TTDs in the untreated tumors. These findings demonstrate that DSC-MRI perfusion methods can be used to non-invasively detect the morphological and functional changes in tumor vasculature that occur in response to dexamethasone treatment. PMID- 15896080 TI - Perineural invasion of squamous cell carcinoma of the lip with occult involvement of the infra-orbital nerve detected by PET-CT and treated with MRI-based IMRT: a case report. AB - A 51 year old male with a history of right facial numbness developed progressive upper lip swelling for one year, but an MRI of the head was unremarkable. A wide local excision of the upper lip was performed and pathology revealed a 1.7 cm mass, poorly differentiated squamous cell carcinoma with perineural invasion. Surgical margins were free of tumor. Two months postoperatively, a hybrid PET-CT of the whole body was performed due to the persistent right facial numbness. The CT portion identified an equivocal lesion at the base of the right orbit correlating to the right infraorbital nerve. However, the PET-CT image revealed avid uptake in this location suggesting perineural invasion which was confirmed with biopsy of the right infraorbital nerve demonstrating carcinoma. Subsequently, the patient was treated with Intensity Modulation Radiation Therapy (IMRT) using MRI fusion for proper delineation of the right infraorbital nerve to its origin in the base of skull. This case exemplifies the superiority of hybrid PET-CT over CT or MRI alone in head and neck imaging which can lead to significant impact on management for patients with head and neck cancer. PMID- 15896081 TI - Pharmacokinetic parameters analyzed from mr contrast enhancement kinetics of multiple malignant and benign breast lesions detected in the same patients. AB - Ninety-nine patients with confirmed breast cancer were reviewed to identify patients who had two confirmed malignant lesions of identical pathology (Group-1, N = 17), and patients who had one malignant lesion and the second benign lesion (Group-2, N = 8). Contrast enhancement kinetics from every lesion was measured and analyzed using three different models to obtain fitting parameters related to up-slope, enhancement amplitude, and wash-out, including Model-1: modified Tofts model (v(p), Ktrans, k(ep)), Model-2: standard Tofts model (Ktrans, k(ep)), and Model-3: a 3-parameter heuristic model (Tc, A, C). By analyzing lesions from same patients, the differences in whole body hemodynamics thus the blood kinetics could be controlled. Two questions were addressed in this study: i) What is the association between pharmacokinetic parameters analyzed from multiple cancers of identical pathology in same patients?; and ii) What is the difference between secondary malignant lesions and secondary benign lesions with reference to the primary cancer? All three models could fit the enhancement kinetics satisfactorily. Regardless of the analysis model the parameter obtained from the primary cancer and the secondary cancer showed significant correlations. In comparison between Group-1 and Group-2 subjects, the wash-out parameter kep in Models-1 and 2 could significantly differentiate benign from malignant lesions, but not the magnitude parameters, Ktrans in Model-2 or the parameter A in Model 3. If analyzed using appropriate models the early up-slope parameters, v(p) in Model-1 and Tc in Model-3, might be able to distinguish between benign and malignant lesions. When more data are available a reference database can be established with the method described in this study, and from which to determine the likelihood of malignancy for each incidental lesion found in preoperative MRI, with reference to the primary cancer. PMID- 15896082 TI - Analysis of spatial correlation between 99mTc-Sestamibi uptake and radiographic breast density. AB - Breast scintigraphy is a technique by which the biological properties of breast lesions can be assessed using an injected radiopharmaceutical. It may be particularly useful for women with radiographically dense breasts, in whose mammograms, lesions are often obscured by breast tissue. We are evaluating a dual modality breast scanner developed at the University of Virginia for its ability to distinguish between benign and malignant lesions. The scanner obtains a digital mammogram and a gamma ray emission image in quick succession with the breast held under mild compression, resulting in a fused image in which structures in the digital mammogram can be directly correlated with those in the scintigram. Our experience has shown that radiopharmaceutical uptake by normal breast tissue can sometimes obscure uptake by small lesions. It would therefore be advantageous to correct for this background uptake if possible. One potential way of accomplishing this is to use the information from the digital mammogram to help predict the background radiopharmaceutical distribution. With this in mind, we retrospectively investigated the degree of spatial correlation between the distribution of background activity and the distribution of radiodense breast tissue in normal breasts. Using a histogram-based analysis, we have quantified the degree of correlation in 16 images obtained from a total of 8 patients. We also used the mammographic images to quantify the radiographic density of each breast. Our results suggest that spatial correlation between areas of high radiopharmaceutical uptake and parenchymal density exists in the most dense regions of the breast for either extremely dense or heterogeneously dense breasts. High correlation was also observed for some homogeneously fatty breasts. In the latter case however, variation in breast thickness appeared to be the cause of the increased correlation. Correlation properties are approximately equal in both right and left breasts for a particular patient, except in cases exhibiting focal radiotracer uptake in a lesion. Although our preliminary results suggest that correlation between radiopharmaceutical uptake and parenchymal density exists, the number of cases thus far is too small for definitive conclusions. In addition, the planar nature of the dual modality scans imposes an inherent limitation on our ability to take into account attenuation of the emitted gamma radiation, which thus constitutes an uncontrolled variable in the correlation analysis. In principle, this problem can be eliminated by 3 dimensional imaging. PMID- 15896083 TI - A clinical interactive technique for MR-CT image registration for target delineation of intracranial tumors. AB - Replacement of current CT-based, three-dimensional (3D) treatment planning systems by newer versions capable of automated multi-modality image registration may be economically prohibitive for most radiation oncology clinics. We present a low-cost technique for MR-CT image registration on a "first generation" CT-based, 3D treatment planning system for intracranial tumors. The technique begins with fabrication of a standard treatment mask. A second truncated mask, the "minimask," is then made, using the standard mask as a mold. Two orthogonal leveling vials glued onto the minimask detect angular deviations in pitch and roll. Preservation of yaw is verified by referencing a line marked according to the CT laser on the craniocaudal axis. The treatment mask immobilizes the patient's head for CT. The minimask reproduces this CT-based angular treatment position, which is then maintained by taping the appropriately positioned head to the MR head coil for MR scanning. All CT and MR images, in DICOM 3.0 format, are entered into the treatment planning system via a computer network. Interactive registration of MR to CT images is controlled by real-time visual feedback on the computer monitor. Translational misalignments at the target are eliminated or minimized by iterative use of qualitative visual inspection. In this study, rotational errors were measured in a retrospective series of 20 consecutive patients who had undergone CT-MR image registration using this technique. Anatomic structures defined the three CT orthogonal axes from which angular errors on MR image were measured. Translational errors at the target isocenter were within pixel size, as judged by visual inspection. Clinical setup using the minimask resulted in overall average angular deviation of 3 degrees +/-2 degrees (mean +/- SD) and translational deviation within the edges of the target volume of typically less than 2 mm. The accuracy of this registration technique for target delineation of intracranial tumors is compatible with practice guidelines. This method, then, provides a cost-effective means to register MR and CT images for target delineation of intracranial tumors. PMID- 15896085 TI - Pancreatic cancer epidemiology. AB - Carcinoma of the exocrine pancreas remains a challenging disease mainly due to advanced stage diagnosis, the early systemic dissemination, aggressive local tumor progression, and subsequent short patient survival. Thus, assessment of incidence markers instead of prevalence indicators is recommended because the time between diagnosis and death is usually very short. Investigations to date have led to the discovery of many rare genes and environmental factors that contribute to pancreatic cancer. However, common genes involved in genetic polymorphisms, and specific risk factors have not been identified. Furthermore, the role of gene-environment interactions on the tumorogenesis and progression of pancreatic cancer need to be further investigated. Given current therapeutic modalities, only early detection of cancer of the pancreas, followed by surgical resection, offers the possibility of lengthening survival time. PMID- 15896084 TI - A review of progress in clinical photodynamic therapy. AB - Photodynamic therapy (PDT) has received increased attention since the regulatory approvals have been granted to several photosensitizing drugs and light applicators worldwide. Much progress has been seen in basic sciences and clinical photodynamics in recent years. This review will focus on new developments of clinical investigation and discuss the usefulness of various forms of PDT techniques for curative or palliative treatment of malignant and non-malignant diseases. PMID- 15896086 TI - Survey of resident education in intensity-modulated radiation therapy. AB - Intensity-modulated radiation therapy (IMRT) has been gaining increasing popularity among practicing physicians in the U.S., but the extent to which radiation oncology residents are taught the principles of this technology and are trained to use IMRT remains unknown. In this paper, we assessed the current level of resident education in IMRT in the United States. Chief residents at all 77 accredited radiation oncology programs were sent a 13-question survey addressing formal didactics and hands-on experience in IMRT. The survey assessed the frequency, subject, and format of IMRT didactics. Questions also addressed the number of IMRT patients and anatomical sites treated, resident involvement in the IMRT process, and the intent of IMRT use. Finally, residents were asked for their opinions on their IMRT education. Sixty-one surveys (79%) were completed. Overall, forty-three respondents (71%) reported receiving formal IMRT didactics, with nearly one-third reporting extensive didactics (> or = 3 lectures/seminars et cetera per year). The most common didactic formats were lectures (95%) and journal clubs (63%), most commonly supervised by physicists (98%). Involvement by physicians and radiobiologists were reported by 63% and 7% of respondents, respectively. Overall, 87% of respondents had hands-on IMRT training, with nearly one-half having treated > 25 patients. The most common sites treated were head and neck (94%) and prostate (81%). Involvement in all aspects of the IMRT process was common, particularly target and tissue delineation (98%) and plan evaluation (93%). Most respondents (79%) with hands-on experience reported receiving formal didactics. However, nearly one-third received no or only minimal formal didactics. The percentage of respondents desiring increased IMRT didactics and hands-on experience were 70% and 47%, respectively. Our results suggest that the great majority of radiation oncology residents in the United States are currently exposed to didactics and hands-on training in IMRT. Areas of potential improvement include increased involvement of physicians and radiobiologists in formal IMRT didactics. PMID- 15896087 TI - The effects of exercise on the storage and oxidation of dietary fat. AB - Obesity has become a worldwide problem of pandemic proportions. By definition, obesity is the accumulation of excess body fat and it represents the long-term results of positive energy and fat balance. The failures in the regulatory mechanisms leading to the development of obesity are still not well understood, but there is growing evidence that exercise is an important element in obesity prevention. Exercise promotes energy/fat balance while providing beneficial alterations to obesity/overweight-related comorbidities and mortality. Also, exercise, in large part, influences whether the fate of dietary fat is storage or oxidation. Many factors including intensity, duration and type (aerobic vs anaerobic) of exercise, energy expended during exercise and individual fitness level impact the amounts of fat oxidised at any given time. Evidence suggests that moderate-intensity exercise yields the most cumulative (during and post exercise) fat grams used for substrate in the average individual. All intensities of exercise, however, promote fat oxidation during the post-exercise period. We suggest that it is the effects of exercise on 24-hour fat balance that are most important in understanding the role of exercise in the prevention of fat accumulation and obesity. PMID- 15896088 TI - Exercise training in obese children and adolescents: current concepts. AB - Childhood obesity has reached epidemic proportions worldwide and is associated with increased cardiovascular mortality and morbidity in adult life. The increase in fat mass in children and adolescents has occurred concomitantly with a decline in reported time for exercise. Evidence suggests that non-physically active children are more likely to become non-physically active adults and that encouraging the development of physical activity habits in children helps establish patterns that continue into adulthood. Dietary treatment of obesity is relatively ineffective in adults and it has been suggested that prevention of obesity in childhood and adolescence should emphasise increased physical activity rather than diet because of fears relating to the adverse effects of inappropriate eating patterns. Despite this, there are very few randomised controlled studies investigating the efficacy of exercise training in obese children or adolescents and many of the extant studies have been poorly controlled and have not specifically stratified the independent effect of exercise versus dietary modification. This review focuses on the well designed controlled trials that have evaluated the effect of exercise training in obese children and adolescents on body composition, haemodynamic and metabolic variables, cardiovascular fitness, muscular strength and vascular function. These studies indicate that, although exercise training does not consistently decrease bodyweight or body mass index, it is associated with beneficial changes in fat and lean body mass, emphasising the importance of comprehensive assessment of body composition in future exercise-training studies. Exercise training improves cardiovascular fitness and muscular strength; however, it seems to have little effect on blood lipid profile or blood pressure in obese young people. Importantly, recent studies have demonstrated that exercise training improves vascular endothelial function, an important surrogate measure that may predict future atherosclerotic risk in obese children and adolescents. Given that improvement in vascular function in these training studies occurred in the absence of changes in lipid fractions, haemodynamic variables or glucose metabolism, exercise appears to have a direct beneficial effect on the vasculature, in addition to its putative benefits through risk-factor modification. PMID- 15896090 TI - Skeletal muscle damage with exercise and aging. AB - Skeletal muscle comprises the largest organ system in the human body and is essential for force generation and movement. Skeletal muscle is subjected to considerable stresses during everyday use. However, muscle has the unique ability to adapt and remodel to provide protection against such stresses. This adaptation occurs at the structural through to the cellular level, which includes changes in transcription of a range of protective proteins. Failure in such processes can be catastrophic. This failure in adaptation is particularly notable in older individuals. Our skeletal muscles become smaller and weaker as we age. This loss of muscle bulk results in a reduced capacity to generate force and results in a loss of the ability to undertake everyday tasks. This article describes the normal adaptive responses of muscle in younger individuals to the stress of various forms of exercise and the implications of a failure of these adaptive responses in the elderly. PMID- 15896089 TI - Interactions of metabolic hormones, adipose tissue and exercise. AB - Physiological and psychological systems work together to determine energy intake and output, and thus maintain adipose tissue. In addition, adipose tissue secretes leptin and cytokines, which induces satiety and has been linked to catecholamines, cortisol, insulin, human growth hormone, thyroid hormones, gonadotropin and lipolysis. Thus, adipose tissue is acted upon by a number of physiological stimuli, including hormones, and simultaneously, is an active component in the regulation of its own lipid content. All of the hormones mentioned above are associated with each other and respond to exercise and exercise training. Thus, exercise is one of the major links between the hormonal modulators of energy intake and output. It appears that the sympathetic nervous system and the catecholamines are key components facilitating the lipolytic activity during exercise. These two neuroendocrine factors directly affect adipose metabolism and metabolic hormones that influence adipose metabolism. Acute low- and moderate-intensity exercise causes hormonal changes that facilitate lipolytic activity. Exercise training reduces these hormonal responses, but the sensitivity to these hormones increases so that lipolysis may be facilitated. Large amounts of adipose tissue blunt the metabolic hormonal responses to exercise, but the sensitivity of these hormones is increased; thus maintaining normal lipolytic activity. Although the physiological role of the endocrine system during exercise and training is significant, other training effects may have as great, or greater influence on lipolytic activity in adipose tissue. PMID- 15896091 TI - The role of biomechanics in maximising distance and accuracy of golf shots. AB - Golf biomechanics applies the principles and technique of mechanics to the structure and function of the golfer in an effort to improve golf technique and performance. A common recommendation for technical correction is maintaining a single fixed centre hub of rotation with a two-lever one-hinge moment arm to impart force on the ball. The primary and secondary spinal angles are important for conservation of angular momentum using the kinetic link principle to generate high club-head velocity. When the golfer wants to maximise the distance of their drives, relatively large ground reaction forces (GRF) need to be produced. However, during the backswing, a greater proportion of the GRF will be observed on the back foot, with transfer of the GRF on to the front foot during the downswing/acceleration phase. Rapidly stretching hip, trunk and upper limb muscles during the backswing, maximising the X-factor early in the downswing, and uncocking the wrists when the lead arm is about 30 degrees below the horizontal will take advantage of the summation of force principle. This will help generate large angular velocity of the club head, and ultimately ball displacement. Physical conditioning will help to recruit the muscles in the correct sequence and to optimum effect. To maximise the accuracy of chipping and putting shots, the golfer should produce a lower grip on the club and a slower/shorter backswing. Consistent patterns of shoulder and wrist movements and temporal patterning result in successful chip shots. Qualitative and quantitative methods are used to biomechanically assess golf techniques. Two- and three-dimensional videography, force plate analysis and electromyography techniques have been employed. The common golf biomechanics principles necessary to understand golf technique are stability, Newton's laws of motion (inertia, acceleration, action reaction), lever arms, conservation of angular momentum, projectiles, the kinetic link principle and the stretch-shorten cycle. Biomechanics has a role in maximising the distance and accuracy of all golf shots (swing and putting) by providing both qualitative and quantitative evidence of body angles, joint forces and muscle activity patterns. The quantitative biomechanical data needs to be interpreted by the biomechanist and translated into coaching points for golf professionals and coaches. An understanding of correct technique will help the sports medicine practitioner provide sound technical advice and should help reduce the risk of golfing injury. PMID- 15896092 TI - Iliotibial band syndrome in runners: innovations in treatment. AB - Iliotibial band syndrome (ITBS) is the most common cause of lateral knee pain in runners. It is an overuse injury that results from repetitive friction of the iliotibial band (ITB) over the lateral femoral epicondyle, with biomechanical studies demonstrating a maximal zone of impingement at approximately 30 degrees of knee flexion. Training factors related to this injury include excessive running in the same direction on a track, greater-than-normal weekly mileage and downhill running. Studies have also demonstrated that weakness or inhibition of the lateral gluteal muscles is a causative factor in this injury. When these muscles do not fire properly throughout the support phase of the running cycle, there is a decreased ability to stabilise the pelvis and eccentrically control femoral abduction. As a result, other muscles must compensate, often leading to excessive soft tissue tightness and myofascial restrictions. Initial treatment should focus on activity modification, therapeutic modalities to decrease local inflammation, nonsteroidal anti-inflammatory medication, and in severe cases, a corticosteroid injection. Stretching exercises can be started once acute inflammation is under control. Identifying and eliminating myofascial restrictions complement the therapy programme and should precede strengthening and muscle re-education. Strengthening exercises should emphasise eccentric muscle contractions, triplanar motions and integrated movement patterns. With this comprehensive treatment approach, most patients will fully recover by 6 weeks. Interestingly, biomechanical studies have shown that faster-paced running is less likely to aggravate ITBS and faster strides are initially recommended over a slower jogging pace. Over time, gradual increases in distance and frequency are permitted. In the rare refractory case, surgery may be required. The most common procedure is releasing or lengthening the posterior aspect of the ITB at the location of peak tension over the lateral femoral condyle. PMID- 15896093 TI - Quality control in the regulation of pharmaceuticals. PMID- 15896094 TI - Willingness to pay for a QALY: theoretical and methodological issues. AB - What is a QALY worth in monetary units? This paper presents the main arguments in the literature regarding the obstacles involved in establishing one unique willingness to pay (WTP) estimate for the value of a QALY. To directly translate QALYs into monetary units, and in this manner translate existing and forthcoming cost-effectiveness analyses (CEA) to cost-benefit analyses (CBA), it is necessary that one unique WTP per QALY can be established irrespective of context-specific characteristics such as severity of illness, magnitude of health gain, patient characteristics, etc. Because CEA and CBA are two methods of economic evaluation that are based on two very different normative perceptions of the role of health versus other goods in society, the task of performing a linear translation from QALYs to WTP is theoretically unattainable. CBA is based on the welfarist perception that the welfare associated with health is measured by way of individual preferences for health outcomes relative to other goods in society. In contrast, CEA is based on the extra-welfarist notion, which focuses on maximising health and not welfare, and suppresses any variation across income/social groups in utility derived from improvements in health. Another obstacle to one unique WTP per QALY value is that marginal utility of income is non-constant, and a function of income level and possibly health status. When marginal utility of income varies across individuals as well as contexts, measuring the value of health in monetary units may result in valuations of health increments that are very different from valuations retrieved had another unit of measure been applied. In conclusion, from a theoretical point of view, establishing one unique WTP cannot be attained. Applying one sole WTP per QALY value will entail overriding individual preferences such as diminishing marginal utility of health and potential differences in the value of incremental health across population groups. However, one problem that can, and should, be overcome when seeking to establish a monetary value for a QALY is the problem of variance in the marginal utility of income. The importance of applying the appropriate perspective when formulating WTP questions to ensure that the marginal utility of income of the respondents equals that of the financiers of the costs invested to produce the health gains should not be overlooked. PMID- 15896095 TI - Effects of antidepressants on quality of life in women with premenstrual dysphoric disorder. AB - This review examines the effects of antidepressant medications on premenstrual dysphoric disorder (PMDD) and the diminished quality of life (QOL) that accompanies the disorder. PMDD is a chronic condition in women that emerges in the second half of the menstrual cycle and remits during the menstrual period. The affective and behavioural symptoms of PMDD adversely affect functioning and QOL to a disabling degree, particularly in the domains of family and personal relationships, work productivity and social activities. The serotonergic antidepressants, specifically the selective serotonin reuptake inhibitors (SSRIs), are effective for PMDD. Continuous and luteal-phase dosing regimens with SSRIs are similarly effective and well tolerated. Treatment of PMDD with a serotonergic antidepressant significantly improves functioning and QOL in all studies that have systematically examined QOL issues in this disorder. Although the data show that PMDD is effectively treated with serotonergic antidepressants and that functional impairment that accompanies the disorder is also improved with treatment, the social and economic burden of PMDD continues to be widely unrecognised. Greater awareness of the effectiveness of treatments and reliable measures of the direct and indirect healthcare costs of the disorder when it remains untreated are needed. PMID- 15896096 TI - Does pharmacogenomics provide an ethical challenge to the utilisation of cost effectiveness analysis by public health systems? AB - Pharmacogenomics promotes an understanding of the genetic basis for differences in efficacy or toxicity of drugs in different individuals. Implementation of the outcomes of pharmacogenomic research into clinical practice presents a number of difficulties for healthcare. This paper aims to highlight one of the unique ethical challenges which pharmacogenomics presents for the utilisation of cost effectiveness analysis by public health systems. This paper contends that pharmacogenomics provides a challenge to fundamental principles which underlie most systems for deciding which drugs should be publicly subsidised. Pharmacogenomics brings into focus the conflict between equality and utility in the context of using cost-effectiveness analysis to aid distribution of a limited national drug budget. PMID- 15896097 TI - Valuing patient and caregiver time: a review of the literature. AB - As healthcare expenditures continue to rise, financial pressures have resulted in a desire for countries to shift resources away from traditional areas of spending. The consequent devolution and reform have resulted in increased care being provided and received within homes and communities, and in an increased reliance on unpaid caregivers. Recent empirical work indicates that costs incurred by care recipients and unpaid caregivers, including time and productivity costs, often account for significant proportions of total healthcare expenditures. However, many economic evaluations do not include these costs. Moreover, when indirect costs are assessed, the methods of valuation are inconsistent and frequently controversial. This paper provides an overview and critique of existing valuation methods. Current methods such as the human capital method, friction cost method and the Washington Panel approach are presented and critiqued according to criteria such as potential for inaccuracy, ease of application, and ethical and distributional concerns. The review illustrates the depth to which the methods have been theoretically examined, and highlights a paucity of research on costs that accrue to unpaid caregivers and a lack of research on time lost from unpaid labour and leisure. To ensure accurate and concise reporting of all time costs, it is concluded that a broad conceptual approach for time costing should be developed that draws on and then expands upon theoretical work to date. PMID- 15896098 TI - The health-related quality of life and economic burden of constipation. AB - Constipation is a prevalent condition that disproportionately affects women and older adults and leads to self-medication and/or medical consultation. It occurs as a result of functional idiopathic causes or secondarily as a result of a variety of factors including dietary and exercise patterns, adverse effects of medication and disease processes. Constipation is often perceived to be a benign, easily treated condition with short-term treatment being relatively straightforward; however, chronic constipation is associated with mild complications that, left untreated, can develop into more serious bowel complaints (faecal impaction, incontinence and bowel perforations) with further implications for healthcare costs and the patient's health-related quality of life (HR-QOL). This review summarises the evidence of the HR-QOL impact and economic burden of constipation on patients. Relatively few studies have systematically explored the HR-QOL and economic impact of constipation; however, the existing evidence suggests that HR-QOL is lower in patients with constipation than in non-constipated individuals, and treatments for constipation improve HR QOL. Additionally, constipation represents an economic burden for the patient and healthcare provider. Resource utilisation associated with the diagnosis and management of constipation is a significant cost driver, whereas constipation prevention programmes have demonstrated cost savings. PMID- 15896099 TI - Economic considerations in the prescribing of third-generation antidepressants. AB - A comprehensive, multinational literature search was conducted of all articles published from 1993 to 2003 regarding the cost effectiveness of antidepressant drugs, with special emphasis on comparing third-generation antidepressants (TGAs) with tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs). Information from the collected articles was entered into a database and then analysed to assess the different approaches to cost effectiveness comparisons for the various classes of antidepressants. Factors examined included direct and indirect costs, treatment endpoints, healthcare cost burden and productivity gains for patients successfully treated for depression. Most model based studies published between 1993 and 2003 supported the cost effectiveness of TGAs compared with TCAs or SSRIs. While the purchase price of TGAs may be greater, cost savings in terms of direct and indirect costs were realised because of the reduced adverse effects of TGAs and subsequent improved patient compliance. Studies based on patient level cost data, however, were less conclusive about the economic benefits of TGAs compared with SSRIs or TCAs. While it may be premature to conclude that TGAs have a significant net economic benefit compared with other antidepressant classes, prescribers and payers may find it helpful when choosing antidepressants for depressed patients to consider that the higher drug prices for TGAs may be offset by savings, in terms of their greater compliance and resultant therapeutic success rates compared with TCAs or SSRIs. Additional research is needed to clarify how cost effectiveness is assessed in different patient populations, such as geriatric patients--who commonly have more co-morbidities and higher total healthcare costs than other patient populations. PMID- 15896100 TI - Economic evaluation of levetiracetam as an add-on therapy in patients with refractory epilepsy. AB - OBJECTIVES: This study provides the results of a cost-effectiveness analysis of levetiracetam as an adjunctive treatment for refractory epilepsy from the Canadian Ministry of Health perspective. The main objective is to estimate the expected cost-effectiveness ratio expressed as the incremental cost per seizure free day gained when using levetiracetam. In addition, this study examines the potential savings that might result by reducing the number of surgical evaluations and surgery when using levetiracetam. METHODS: A 1-year dose escalation decision-tree model comparing levetiracetam plus standard therapy with standard therapy alone was designed in order to combine probability, resource use and unit cost data (1999 Canadian dollars [$Can]). The short-term outcomes were derived from three phase III randomised, double-blind, placebo-controlled trials performed in 904 patients, aged 16-70 years, with at least 1 year history of epilepsy, two to four partial seizures per month, and receiving a maximum of two classic antiepileptic drugs. RESULTS: The average gain in seizure-free days attributed to levetiracetam was 19 days per patient per year and the incremental cost-effectiveness ratio (ICER) for levetiracetam add-on in the base-case scenario was $Can80.7 per seizure-free day gained per patient per year. Moreover, when surgical investigation and surgery are considered in the model, the use of levetiracetam may be dominant, with substantial savings to the overall healthcare budget. All univariate sensitivity analyses show that the model was robust to the assumptions made. CONCLUSIONS: The economic analysis presented in this paper suggests, given a wide range of assumptions, that the increased cost of treating patients (with refractory epilepsy) with levetiracetam may be partially offset by a reduction in other direct medical costs (from the Canadian Ministry of Health perspective), as a consequence of an increase in the number of seizure-free days. Moreover, potential cost savings may be foreseen when it is assumed that levetiracetam may reduce the number of candidates for surgical evaluation and surgery through a reduction of seizure frequency. PMID- 15896101 TI - Reductions in anaemia and fatigue are associated with improvements in productivity in cancer patients receiving chemotherapy. AB - OBJECTIVE: Cancer-related anaemia is associated with fatigue that adversely affects patients' everyday functioning and wellbeing. We explore the impact of fatigue on patient productivity and caregiver burden. METHODS: The analyses are based on data from a randomised, open-label, active-controlled, dose-finding trial of darbepoetin alfa among solid-tumour cancer patients with anaemia, who are receiving chemotherapy. Fatigue is assessed with the Functional Assessment of Cancer Therapy (FACT)-Fatigue subscale score. Productivity and caregiver outcomes include time (hours) missed from usual activities, amount of assistance (hours) needed from others, overall ability to perform desired activities and ability to perform family responsibilities. These outcomes are assessed at baseline and the end of the 12-week treatment period. ANOVA and linear regression models are used to evaluate associations. RESULTS: Patients (n=300) were aged 61 years on average, with a mean (SD) baseline haemoglobin of 9.9 (0.9) g/dL. FACT-Fatigue subscale score improvements were significantly (p=0.003) associated with haemoglobin improvements. Over a 2-week period, after controlling for age, sex and disease progression, one-point improvements in FACT-Fatigue subscale scores corresponded to a 1-hour (95% CI 0.5, 1.5) gain in productive time, 0.7-hour (95% CI 0.4, 1.0) reduction in caregiver time and 1.6% (95% CI 1.4, 1.7) improvement in overall activity. CONCLUSIONS: Reducing fatigue is associated with gains in productive time, reductions in caregiver burden and enhanced ability to perform activities. These outcomes may have broader implications for patients' wellbeing and for the societal impact of cancer-related fatigue and anaemia. PMID- 15896102 TI - Cost effectiveness of imatinib compared with interferon-alpha or hydroxycarbamide for first-line treatment of chronic myeloid leukaemia. AB - OBJECTIVE: To evaluate the cost utility of imatinib compared with interferon (IFN)-alpha or hydroxycarbamide (hydroxyurea) for first-line treatment of chronic myeloid leukaemia. DESIGN AND SETTING: A cost-utility (Markov) model within the setting of the UK NHS and viewed from a health system perspective was adopted. Transition probabilities and relative risks were estimated from published literature. Costs of drug treatment, outpatient care, bone marrow biopsies, radiography, blood transfusions and inpatient care were obtained from the British National Formulary and local hospital databases. Costs (pound, year 2001-03 values) were discounted at 6%. Quality-of-life (QOL) data were obtained from the published literature and discounted at 1.5%. The main outcome measure was cost per QALY gained. Extensive one-way sensitivity analyses were performed along with probabilistic (stochastic) analysis. RESULTS: The incremental cost-effectiveness ratio (ICER) of imatinib, compared with IFNalpha, was pound26,180 per QALY gained (one-way sensitivity analyses ranged from pound19,449 to pound51,870) and compared with hydroxycarbamide was pound86,934 per QALY (one-way sensitivity analyses ranged from pound69,701 to pound147,095) [ pound1=$US1.691=euro1.535 as at 31 December 2002].Based on the probabilistic sensitivity analysis, 50% of the ICERs for imatinib, compared with IFNalpha, fell below a threshold of approximately pound31,000 per QALY gained. Fifty percent of ICERs for imatinib, compared with hydroxycarbamide, fell below approximately pound95,000 per QALY gained. CONCLUSIONS: This model suggests, given its underlying data and assumptions, that imatinib may be moderately cost effective when compared with IFNalpha but considerably less cost effective when compared with hydroxycarbamide. There are, however, many uncertainties due to the lack of long term data. PMID- 15896103 TI - On pharmacogenomics and cost-effectiveness analysis at the individual level. PMID- 15896108 TI - [Probiotics: fantasy or reality?]. PMID- 15896109 TI - [Study of the influence of dietary yogurt in an allergic population]. AB - BACKGROUND: Prevalence of allergic diseases has been increased in last years; new alternative therapies have been employed. Beneficial effects of probiotics have been described in some pathologies such as tumors, diarrhea, and allergic disease. The aim of our work was to describe nutritional status and influence of probiotic consumption in allergic population. PATIENTS AND METHODS: 44 allergic patients were selected, in all patients were recorded; sex, age, place of residence, anthropometric evaluation, dietary questionnaire and allergic variables (rush, blood levels of Ig E, and number of crisis in a year). RESULTS: 18 patients (40.9%) were females and 26 (59.1%) males with an average age of 24.5 (10.3) years. Most of patients (20.5%) are located in percentile P25-50, showing a good nutritional status. 16 patients consumed probiotics (27.1%), with an average of consumption per week 1.79 (3.16) and an average amount per week (mg ml) (593.4 (461.9). The number of crisis year was 1.44 (1.8), average value of Ig E was 35.68 (31.93) UI/L and rush 10.09 (2.8) mm. Patients with a consumption of probiotics higher than 593 mg/week showed a IOW rush diameter (10.12(0.8) mm vs 8.85 (1.1) mm; p < 0.05), without statistical differences in other variables. CONCLUSION: Consumption of probiotics in allergic patients could be beneficial. Further studies with more patients and intervention designs will be necessary to analyze this relations. PMID- 15896110 TI - [Nosocomial infection by multiresistant pathogens during one year in a secondary hospital: clinical an epidemiological analysis]. AB - BACKGROUND: Currently growing medical and social significance of nosocomial infection by multiresistant pathogens (NIMP) prompted us to establish its incidence, nosology, presenting forms in admission areas, and mortality in a secondary hospital, Lleida (Spain). METHOD: For that purpose, we analyzed the first year experience of a unit for the control of nosocomial infection (NI) created in our hospital. From January to December 2000, 79 patients with a NIMP admitted to the University Hospital Arnau de Vilanova entered in this prospective, descriptive study. RESULTS: The overall annual incidence of NIMP was 4.0 per 103 patients admitted. Acinetobacter baumannii showed the highest individual rate of incidence, particularly, at the Intensive Care Unit (15.4 per 103 patients admitted; p < 0.001). By nosologies, infection prevailed over colonization (69.6% vs 30.4%; p < 0.001). Mean hospital stay length increased in colonized patients (38.9 days). Finally, overall mortality was high (29.1%); again, A. baumannii was the agent most frequently detected in death cases (66.6%; p < 0.001). CONCLUSIONS: Surveillance and control measures are required for the prevention of NIMP. Incidence studies how this, can be useful to create a database to establish the distribution and occurrence of NI, including the detection of multiresistant pathogen outbreaks. PMID- 15896175 TI - Indigenous health: partners in healing. AB - The past 12 months have brought considerable changes that affect the lives of Australia's Indigenous people. PMID- 15896176 TI - "Mutual" obligation in Indigenous health: can shared responsibility agreements be truly mutual? AB - Shared responsibility agreements between the Australian Government and Indigenous communities are based on a concept of mutual obligation but have overtones of paternalism and imposition. The nature and extent of choice in any such agreements need to be established. PMID- 15896177 TI - Bridging the treatment gap for Indigenous Australians. Demands for efficiency should not be met at the expense of equity. AB - Demands for efficiency should not be met at the expense of equity. PMID- 15896178 TI - Rates of percutaneous coronary interventions and bypass surgery after acute myocardial infarction in Indigenous patients. AB - OBJECTIVE: To compare rates of percutaneous coronary interventions (PCI) and bypass surgery after acute myocardial infarction (AMI) in Indigenous and non Indigenous patients. DESIGN: Cohort study of public-sector patients who were followed up for 1 year using administrative hospital data. PARTICIPANTS AND SETTING: We followed up 14 683 public-sector patients admitted to Queensland hospitals for AMI between 1998 and 2002. Of these, 558 (3.8%) identified as Indigenous. OUTCOME MEASURES: Rates of PCI and bypass surgery, adjusted for differences between the Indigenous and non-Indigenous cohorts according to age, sex, socioeconomic status, remote residence, hospital characteristics, and comorbidities. RESULTS: The adjusted rate for PCI during the index admission was significantly lower by 39% (rate ratio [RR], 0.61; 95% CI, 0.38-0.98) among Indigenous versus non-Indigenous patients with AMI; the adjusted rate for subsequent PCI was significantly lower by 28% (RR, 0.72; 95% CI, 0.54-0.96). Adjusted rates for bypass surgery were similar in the two cohorts. For any coronary procedure (ie, PCI or bypass surgery), the adjusted rate was significantly lower by 22% (RR, 0.78; 95% CI, 0.64-0.94) among Indigenous patients with AMI. Diabetes, chronic renal failure, pneumonia, and chronic rheumatic fever were at least twice as common among Indigenous patients with AMI as in the rest of the cohort, and chronic bronchitis and emphysema and heart failure were at least 60% more common. If a patient had at least one comorbidity, then their probability of having a coronary procedure was reduced by 40%. CONCLUSIONS: There are likely to be several reasons for the lower rates of coronary procedures among Indigenous patients, but their high rates of comorbidities and the association of comorbidities with lower procedure rates was an important finding. As investment in primary care can reduce the prevalence and severity of comorbidities, we suggest that adequate primary health care is a prerequisite for effective specialist care. PMID- 15896179 TI - Impact of a collaborative shared antenatal care program for urban Indigenous women: a prospective cohort study. AB - OBJECTIVES: To evaluate the impact of a community-based, collaborative, shared antenatal care intervention (the Mums and Babies program) for Indigenous women in Townsville. DESIGN AND PARTICIPANTS: Prospective cohort study of women attending Townsville Aboriginal and Islander Health Service (TAIHS) for shared antenatal care with a singleton Indigenous birth between 1 January 2000 and 31 December 2003 (456 women; the MB group), compared with a historical control group of 84 women who attended TAIHS for antenatal care before the intervention between 1 January 1998 and 30 June1999, and a contemporary control group of 540 women who had a singleton birth at Townsville Hospital between 1 January 2000 and 30 June 2003, but did not attend TAIHS for antenatal care. INTERVENTION: Integration of previously autonomous service providers delivering shared antenatal care from TAIHS. MAIN OUTCOME MEASURES: Patterns of antenatal visits, proportion of women undertaking key antenatal screening, and perinatal outcomes. RESULTS: The number of Indigenous women who entered the MB program and gave birth at Townsville Hospital rose from 23.8% in 2000 to 61.2% in 2003. The number of antenatal care visits per pregnancy increased from three (interquartile [IQ] range, 2-6) in the historical control group to seven (IQ range, 4-10) in the MB group (P < 0.001). 88% of women in the MB group had at least one ultrasound. About 90% of all women attending for antenatal care were screened for sexually transmitted infections. In the MB group, there was a significant reduction in preterm births compared with the contemporary control group (8.7% v 14.3%, P < 0.01). There was no significant reduction in the prevalence of low birthweight births or perinatal mortality. CONCLUSION: A community-based collaborative approach to shared antenatal care services increased access to antenatal care and was associated with fewer preterm births among Indigenous women in Townsville. The model may be adaptable in other urban centres with multiple antenatal care providers and significant numbers of Indigenous people across Australia. PMID- 15896180 TI - Sustaining remote-area programs: retinal camera use by Aboriginal health workers and nurses in a Kimberley partnership. AB - OBJECTIVE: To describe how a novel program of diabetic retinopathy screening was conceived, refined and sustained in a remote region over 10 years, and to evaluate its activities and outcomes. DESIGN: Program description; analysis of regional screening database; audit of electronic client registers of Aboriginal community controlled health services (ACCHSs). SETTING AND PARTICIPANTS: 1318 Aboriginal and 271 non-Aboriginal individuals who underwent retinal screening in the 5 years to September 2004 in the Kimberley region of north-west Australia; 11 758 regular local Aboriginal clients of Kimberley ACCHSs as at January 2005. MAIN OUTCOME MEASURES: Characteristics of clients and camera operators, prevalence of retinopathy, photograph quality, screening intervals and coverage. RESULTS: Among Aboriginal clients, 21% had diabetic retinopathy: 19% with non-proliferative retinopathy, 1.2% with proliferative retinopathy, and 2.8% with maculopathy. Corresponding figures for non-Aboriginal clients were 11%, 11%, 0 and 0.4%, respectively. Photograph quality was generally high, and better for non Aboriginal clients, younger Aboriginal clients and from 2002 (when mydriatic use became universal). Quality was not related to operator qualifications, certification or experience. Of 718 regular Aboriginal clients with diabetes on local ACCHS databases, 48% had a record of retinal screening within the previous 18 months, and 65% within the previous 30 months. CONCLUSIONS: Screening for diabetic retinopathy performed locally by Aboriginal health workers and nurses with fundus cameras can be successfully sustained with regional support. Formal certification appears unnecessary. Data sharing across services, client recall and point-of-care prompts generated by electronic information systems, together with policies making primary care providers responsible for care coordination, support appropriate timely screening. PMID- 15896181 TI - Point-of-care testing of HbA1c and blood glucose in a remote Aboriginal Australian community. AB - OBJECTIVES: To assess the accuracy of point-of-care (POC) measurements of capillary blood glucose and glycosylated haemoglobin (HbA(1c)) levels in a remote Aboriginal community with high diabetes prevalence. DESIGN: Cross-sectional study comparing POC capillary glucose and HbA(1c) results with those from corresponding venous samples measured in a reference laboratory. PARTICIPANTS AND SETTING: 152 residents aged 11-76 years (representing 76% of population aged over 11 years) had POC glucose measurement in November 2003; 88 with POC glucose level > or = 5.0 mmol/L, or self-reported diabetes, had POC HbA(1c) and laboratory glucose and HbA(1c) measurements. MAIN OUTCOME MEASURES: POC fasting capillary levels of glucose (HemoCue Glucose 201 analyser, Medipac Scientific, Sydney) and HbA(1c) (DCA 2000+ analyser, Bayer Australia, Melbourne); correlation and mean difference between capillary POC and venous blood laboratory measurements of glucose and HbA(1c). RESULTS: Mean and median POC capillary glucose levels were 7.99 mmol/L and 6.25 mmol/L, respectively, while mean and median laboratory venous plasma glucose concentrations were 7.63 mmol/L and 5.35 mmol/L. Values for POC capillary HbA(1c) and laboratory HbA(1c) were identical: mean, 7.06%; and median, 6.0%. The correlation coefficient r for POC and laboratory results was 0.98 for glucose and 0.99 for HbA(1c). The mean difference in results was 0.36 mmol/L for glucose (95% CI, 0.13-0.62; limits of agreement [LOA], - 2.07 to 2.79 mmol/L; P = 0.007) and < 0.01% for HbA(1c) (95% CI, - 0.07% to 0.07%; LOA, - 0.66% to 0.66%; P = 0.95), respectively. CONCLUSIONS: POC capillary HbA(1c) testing, in particular, offers an accurate, practical, community-friendly way of monitoring diabetes in rural and remote clinical settings. POC capillary glucose results should be confirmed by a laboratory test of venous plasma if the results are likely to significantly influence clinical decisions. PMID- 15896182 TI - The Royal Darwin Hospital as a centre of excellence for clinical training in Aboriginal health: still a dream. AB - Strong leadership is required to overcome the bureaucratic and financial obstacles. PMID- 15896183 TI - Zinc and vitamin A supplementation in Australian Indigenous children with acute diarrhoea: a randomised controlled trial. AB - OBJECTIVE: To evaluate the role of zinc and vitamin A supplementation in the recovery of Indigenous children hospitalised for acute diarrhoea. DESIGN: A randomised controlled 2 by 2 factorial trial of supplementation with zinc and vitamin A. SETTING AND PARTICIPANTS: Aboriginal children (aged < 11 years) hospitalised for acute diarrhoea at Alice Springs Hospital, Northern Territory, April 2001-July 2002. MAIN OUTCOME MEASURES: Duration of diarrhoeal illness; re admission for diarrhoeal illness within 120 days. RESULTS: Our study involved 392 Aboriginal children with 436 episodes of diarrhoea. Supplementation with zinc, vitamin A, or combined zinc and vitamin A had no significant effect on duration of diarrhoea or rate of re-admission compared with placebo. Median diarrhoea duration after starting supplementation was 3.0 days for the vitamin A and zinc supplemented and placebo groups (P values 0.25 and 0.69, respectively). The number of re-admissions did not differ significantly between those receiving vitamin A or zinc and the relevant placebo groups (relative risk [95% CI], 1.2 [0.7-2.1] and 1.3 [0.8-2.1], respectively). CONCLUSION: Vitamin A and zinc supplementation may not be indicated for in-hospital management of acute diarrhoeal disease in Aboriginal children living in remote areas. This finding may not apply to children with malnutrition, for whom other studies suggest a benefit. Larger trials incorporating more comprehensive data on the vitamin A and zinc status as well as nutritional status of study populations might help to explain the different results in different populations. PMID- 15896184 TI - Tropical sprue in Far North Queensland. PMID- 15896185 TI - Dreamtime and awakenings: facing realities of remote area Aboriginal health. AB - It takes humility to walk along the path towards better Aboriginal health. PMID- 15896186 TI - Fish traps -- a significant part of our health and wellbeing. PMID- 15896187 TI - The Dr Ross Ingram Memorial Essay Competition 2005. Fish traps--a significant part of our health and wellbeing. PMID- 15896188 TI - Prevalence of Helicobacter pylori in Indigenous Western Australians: comparison between urban and remote rural populations. PMID- 15896189 TI - Optimizing drug delivery systems using systematic "design of experiments." Part II: retrospect and prospects. AB - The first study on application of Design of Experiments (DoE) in optimizing drug formulation appeared in 1967. Since then the number of literature reports on the use of DoE optimization in development of drug delivery technologies has been piling up steadily. Such systematic techniques find their use in every type of conventional dosage form and modern drug delivery system. The drug delivery devices investigated for optimization using response surface methodology include controlled release compressed matrices, microparticulates, macroparticulates, vesicular systems, floating systems, bioadhesive systems, semisolids, transdermals, and inhalations. The optimized processes mainly include extrusion spheronization, pelletization, microencapsulation, coating, granulation, and tableting. Part I of this article [Crit Rev Ther Drug Carrier Syst 2005; 22(1):27 106] dealt with the salient steps involved in the DoE optimization methodology using diverse experimental designs. Part II deals with various retrospective literature findings as well as the prospective application of such DoE procedures while optimizing varied drug delivery technologies. A vast account of various DoE reports on optimization of diverse drug delivery system and processes along with the critical graphical analysis of various designs, input, and response variables have been presented here in a categorical form. Such an explicit and updated review on drug delivery optimization has not been published anywhere else in the recent past. PMID- 15896190 TI - Advances in buccal drug delivery. AB - The buccal route offers an attractive alternative for systemic drug delivery of drugs because of better patient compliance, ease of dosage form removal in emergencies, robustness, and good accessibility. Use of buccal mucosa for drug absorption was first attempted by Sobrero in 1847, and since then much research was done to deliver drugs through this route. Today, research is more focused on the development of suitable delivery devices, permeation enhancement, and buccal delivery of drugs that undergo a first-pass effect, such as cardiovascular drugs, analgesics, and peptides. In addition, studies have been conducted on the development of controlled or slow release delivery systems for systemic and local therapy of diseases in the oral cavity. In this review, the anatomy and physiology of buccal mucosa, followed by discussion of recent literature on the buccal permeation enhancement, and pathways of enhancement for various molecules are detailed. In addition, bioadhesion theories from historic perspective and current status are discussed. The various dosage forms on the market and in different stages of development are also reviewed. PMID- 15896191 TI - Artificial nerve conduits in peripheral-nerve repair. AB - Injuries to the nervous system are the result of mechanical, thermal, chemical or congenital pathologies and, if function is not restored, they lead to loss of muscle function, pain and impaired sensation. Current treatment modalities essentially coapt the two nerves ends together or place a nerve graft between the cut ends. However, clinical results have never been optimal, and therefore a quest for better options has taken place. In this review article we look at the synthetic and biomimetic options currently being tested as potential nerve grafts. PMID- 15896192 TI - Structural changes in the airways in asthma: observations and consequences. AB - Structural changes reported in the airways of asthmatics include epithelial fragility, goblet cell hyperplasia, enlarged submucosal mucus glands, angiogenesis, increased matrix deposition in the airway wall, increased airway smooth muscle mass, wall thickening and abnormalities in elastin. Genetic influences, as well as fetal and early life exposures, may contribute to structural changes such as subepithelial fibrosis from an early age. Other structural alterations are related to duration of disease and/or long-term uncontrolled inflammation. The increase in smooth muscle mass in both large and small airways probably occurs via multiple mechanisms, and there are probably changes in the phenotype of smooth muscle cells, some showing enhanced synthetic capacity, others enhanced proliferation or contractility. Fixed airflow limitation is probably due to remodelling, whereas the importance of structural changes to the phenomenon of airways hyperresponsiveness may be dependent on the specific clinical phenotype of asthma evaluated. Reduced compliance of the airway wall secondary to enhanced matrix deposition may protect against airway narrowing. Conversely, in severe asthma, disruption of alveolar attachments and adventitial thickening may augment airway narrowing. The encroachment upon luminal area by submucosal thickening may be disadvantageous by increasing the risk of airway closure in the presence of the intraluminal cellular and mucus exudate associated with asthma exacerbations. Structural changes may increase airway narrowing by alteration of smooth muscle dynamics through limitation of the ability of the smooth muscle to periodically lengthen. PMID- 15896193 TI - Eicosanoids: mediators and therapeutic targets in fibrotic lung disease. AB - Fibrosis is a common end-stage sequella of a number of acute and chronic lung diseases. Current concepts of pathogenesis implicate dysregulated interactions between epithelial cells and mesenchymal cells. Although investigative efforts have documented important roles for cytokines and growth factors in the pathogenesis of fibrotic lung diseases, these observations have not as yet been translated into efficacious therapies, and there is a pressing need for new pathogenetic insights and therapeutic approaches for these devastating disorders. Eicosanoids are lipid mediators derived from arachidonic acid, the most studied of which are the prostaglandins and leukotrienes. Although they are primarily known for their roles in asthma, pain, fever and vascular responses, present evidence indicates that eicosanoids exert relevant effects on immune/inflammatory, as well as structural, cells pertinent to fibrogenesis. In general, leukotrienes promote, whereas prostaglandin E(2) opposes, fibrogenic responses. An imbalance of eicosanoids also exists in pulmonary fibrosis, which favours the production of leukotrienes over prostaglandin E(2). This review highlights the role of this imbalance in the evolution of fibrotic lung disease, discusses the mechanisms by which it may arise and considers approaches for therapeutic targeting of eicosanoids in these conditions. PMID- 15896194 TI - NMR structural analysis of a peptide mimic of the bridging sheet of HIV-1 gp120 in methanol and water. AB - gp120 is a subunit of the Env (viral envelope protein) of HIV-1. The protein consists of inner and outer domains linked by a bridging sheet. Several gp120 residues that bind the neutralizing antibody 17b as well as the cellular co receptor CCR5 (CC chemokine receptor 5), are located in the bridging sheet. Peptides that mimic the 17b-binding regions of gp120 would be useful potential immunogens for the generation of neutralizing antibodies against HIV-1. Towards this end, a 26-residue, four-stranded beta-sheet peptide was designed on the basis of the structure of the bridging sheet, and its structure was characterized in methanol by NMR. In methanol, amide and alpha-proton resonances were well resolved and dispersed. A number of interstrand NOEs (nuclear Overhauser effects) were observed, providing good evidence for multiple turn beta-hairpin structure. NOEs also provided good evidence for all Xxx-D-Pro bonds in the trans configuration and all three turns formed by a two residue D-Pro-Gly segment to be of type II' turn. The structure conforms well to the designed four-stranded beta sheet structure. Approx. 20% of the peptide was estimated to adopt a folded conformation in water, as evidenced by CD spectroscopy. This was consistent with smaller, but still significant, downfield shifts of C(alpha)H protons relative to random-coil values. A second peptide was designed with two disulphide bonds to further constrain the peptide backbone. While structured in methanol, this peptide, like the previous one, also exhibits only partial structure formation in water, as evidenced by CD spectroscopy. PMID- 15896195 TI - External mechanical force as an inhibition process in kinesin's motion. AB - We analysed published force-velocity data for kinesin using classical Michaelis Menten kinetic theory and found that the effect of force on the stepping rate of kinesin is analogous to the effect of a mixed inhibitor in classical inhibition theory. We derived an analytical expression for the velocity of kinesin (the stepping rate, equal to the ATP turnover rate) as a function of ATP concentration and force, and showed that it accurately predicts the observed single molecule stepping rate of kinesin under a variety of conditions. PMID- 15896196 TI - NPC1 late endosomes contain elevated levels of non-esterified ('free') fatty acids and an abnormally glycosylated form of the NPC2 protein. AB - NPC (Niemann-Pick type C) disease is a rare lipidosis characterized by the accumulation of LDL (low-density lipoprotein)-derived non-esterified cholesterol in the E/L (endosomal/lysosomal) system. The gene products that are responsible for the two NPC complementation groups are distinct and dissimilar, yet their cellular and disease phenotypes are virtually indistinguishable. To investigate the relationship between NPC1 and NPC2 and their potential role in NPC disease pathogenesis, we have developed a method for the rapid and efficient isolation of late endocytic vesicles from mouse liver by magnetic chromatography. Late endosomes from Wt (wild-type) and NPC1 mice were found to differ not only in their cholesterol and sphingomyelin content, as expected, but also in their non esterified ('free') fatty acid content, with NPC1 vesicles showing an approx. 7 fold increase in non-esterified fatty acid levels compared with Wt vesicles. Furthermore, we show that the NPC2 protein is in an incompletely deglycosylated form in NPC1 late endosomes by a mechanism that is specific to the NPC2 protein and not a global aberration of protein glycosylation/deglycosylation or trafficking, since NPC2 secreted from NPC1 cells is indistinguishable from that secreted from Wt cells. Also, a greater proportion of the normally soluble cellular NPC2 protein partitions with detergent-insoluble late endosomal internal membrane domains in NPC1 vesicles. In addition, we show that, although a small amount of the NPC2 protein associates with these membranes in Wt vesicles, this localization becomes much more pronounced in NPC1 vesicles. These results suggest that the function of the NPC2 protein may be compromised as well in NPC1 endosomes, which might explain the paradoxical phenotypic similarities of the two NPC disease complementation groups. PMID- 15896197 TI - The impact of eicosanoids on the crosstalk between innate and adaptive immunity: the key roles of dendritic cells. AB - The innate immune response is essentially the first line of defense against an invading pathogen. Through specialized receptors, known as pattern recognition receptors, especially Toll-like receptors, specialized cells of myeloid origin, including macrophages and dendritic cells (DCs) are able to phagocytose microorganisms and induce an innate inflammatory response. Although B and T lymphocytes recognize tissue antigens with high specificity, they are unable to initiate immune responses. The decision to activate an appropriate immune response is made by unique DC, the most professional antigen-presenting cells (APCs) which control the responses of several types of lymphocytes and play central role in the transition between innate and adaptive immunity. Increased secretion of inflammatory endogenous mediators such as cytokines and arachidonic acid-derived lipid mediators, also termed eicosanoids, can activate APC, particularly DC, which in turn induce an adaptive immune response. There is an increasing evidence that eicosanoids play an important role in connecting innate and adaptive immunity by acting on cells of both systems. Prostanoids, a major class of eicosanoids, have a great impact on inflammatory and immune responses. PGE(2) is one of the best known and most well-characterized prostanoids in terms of immunomodulation. Although cytokines are known as key regulators of immunity, eicosanoids, including PGE(2), PGD(2), LTB(4), and LTC(4), may also affect cells of immune system by modulating cytokine release, cell differentiation, survival, migration, antigen presentation, and apoptosis. By acting on various aspects of immune and inflammatory reactions, these lipid mediators emerge as key regulators of the crosstalk between innate and adaptive immunity. PMID- 15896198 TI - Role of NK cells in mismatched unrelated haematopoietic stem-cell transplantation: fact or fiction? AB - The role of alloreactive NK cells in haematopoietic stem-cell transplantation is a controversial topic. Different studies come up with different conclusions. This is mainly due to heterogeneous patient populations, differences in the sources of haematopoietic stem cells and various conditioning regimens. International collaboration is essential to reveal in which situation NK cell alloreactivity plays a role and, if so, whether it is beneficial or detrimental for patient survival. PMID- 15896199 TI - Natural killer cell HLA-C epitopes and killer cell immunoglobulin-like receptors both influence outcome of mismatched unrelated donor bone marrow transplants. AB - Matching of donor and recipient for the class I human leukocyte antigen-C (HLA-C) encoded natural killer (NK) epitopes has been reported to influence stem-cell (SC) graft outcome, but a consistent picture has not yet emerged. We have analyzed transplant outcome in 104 unrelated SC grafts in relation to NK epitope (C1 and C2) matching and donor killer cell immunoglobulin-like receptor (KIR) genotype. NK epitope mismatching in the rejection direction was strongly associated with an increased probability of rejection subsequent to engraftment. The prevalence of grades III-IV acute graft-vs-host disease (GVHD) was significantly higher and occurred significantly earlier when there was NK epitope mismatching in the GVH direction. Higher transplant-related mortality and lower disease-free survival rates were associated with epitope mismatching regardless of the mismatch direction. A greater number of KIR receptors, both activating and inhibitory, in the donor protected against grades III-IV GVHD and improved survival. PMID- 15896200 TI - Complete coding sequences and haplotypic associations of HLA-B*0707, -B*1524, B*4405, -B*4802, -DRB1*0409, -DRB1*0411, -DRB1*1115, -DRB1*1305, and the novel allele -DRB1*0709. Group-specific amplification of cDNA from DRB1 alleles associated to DRB3 and DRB4. AB - We present here the characterization of the complete coding sequences, previously unavailable, of the human leukocyte antigen (HLA) alleles B*0707, B*1524, B*4405, B*4802, DRB1*0409, DRB1*0411, DRB1*1115, DRB1*1305, and that of a new allele, DRB1*0709. For the isolation of cDNA from the DRB1 gene, we designed a novel set of polymerase chain reaction (PCR) primers that makes it possible to amplify separately the groups of DRB1 alleles associated to each of the DRB3 and DRB4 loci. The primary structures, functional features, evolutionary relationships, haplotypic associations, and population distributions of each of the nine HLA-B and -DRB1 alleles reported here are reviewed. PMID- 15896201 TI - A11 Tetramer-assisted characterization of Rta-specific CD8+ T-cell responses in healthy virus carriers. AB - HLA Class I-restricted CD8(+) T-cell responses are believed to play an important role in controlling Epstein-Barr virus (EBV) infection, which has been consistently associated with nasopharyngeal carcinoma (NPC). Immediate early transactivator Rta of EBV has been shown to be associated with the reactivation of EBV from latency and drive the lytic cascade of EBV and comprise an important target for EBV-specific cellular cytotoxicity. Furthermore, BRLF1 is specifically expressed in NPC tumor cells. The protein product of BRLF1, Rta, could then be considered as a NPC tumor antigen. Therefore, cellular immunity against Rta represents a very important part of the immunity against NPC, as they should prevent the replication of EBV. In the present study, Rta-specific CD8(+) T-cell responses in healthy virus carriers were characterized by using A1101 tetramer containing the known Rta epitope ATIGTAMYK (134-142). We clearly showed A1101/ATIGTAMYK tetramer-reactive CD8(+) T cells in the circulation of healthy virus carriers, ranging from 2.13 to 9.03%. We then studied the expression of perforin and interferon-gamma (IFN-gamma) secretion in these Rta-specific T cells. Our study demonstrated that Rta-specific T cells are capable of IFN-gamma production and nearly 90% of the Rta-specific CD8(+) T cells expressed perforin. Presumably, these are the cells that play an important role in determining the initiation of the lytic cycle or the clearance of EBV. PMID- 15896202 TI - Interleukin-18-promoter polymorphisms are not relevant in rheumatoid arthritis. AB - Interleukin-18 (IL-18), a member of the IL-1 family, is known to play a relevant role in rheumatoid arthritis (RA) physiopathology mainly by promoting the inflammatory response. The aim of this work was to investigate the possible implication of two single-nucleotide polymorphisms (SNPs) [-607 A/C (rs1946518) and -137 G/C (rs187238)] within the IL-18-promoter region in RA predisposition and clinical course. A total of 362 unrelated RA patients and 339 healthy controls were genotyped using a real-time polymerase chain reaction (PCR) method for the -607 A/C SNP and a sequence-specific PCR method (PCR-SSP) for the -137 G/C polymorphism. No statistically significant differences were observed for both -607 and -137 IL-18-promoter polymorphisms between RA patients and controls, considering either allelic or genotypic frequencies. In addition, no association was found with the haplotypes inferred by the two polymorphisms and RA susceptibility. This was also the case when RA patients were stratified according to sex, age at the onset of the disease, rheumatoid factor status, and extraarticular manifestations. Our data suggest that -607 A/C (rs1946518) and 137 G/C (rs187238) polymorphisms within the IL-18-promoter region do not play a major role in RA predisposition. PMID- 15896203 TI - Sequence of the canine major histocompatibility complex region containing non classical class I genes. AB - We have sequenced a segment of 150,102 nucleotides of canine major histocompatibility complex (MHC) DNA, corresponding to the junction of the class I and class III regions. The distal portion contained five class III genes including two tumor necrosis factor genes and the proximal portion contained five genes or pseudogenes belonging to the class I region. The order of the class III region genes was conserved as in the porcine and human MHC regions. The order of the class Ib loci from the proximal side outwards was DLA-53, DLA-12a, DLA-64, stress-induced phosphoprotein-1, followed by DLA-12. Only DLA-64 and DLA-12 display an overall predicted protein sequence compatible with the expression of membrane-anchored glycoproteins. The other class 1b loci do not appear to be functional by sequence analysis. In all, these 10 genes spanned 24% of the total sequence. The remaining 76% comprised of a number of non-coding and repetitive DNA elements including long interspersed nuclear element (LINE) fragments, short interspersed nuclear elements (SINE), and microsatellites. PMID- 15896204 TI - Distribution of killer cell immunoglobulin-like receptor genes in the Chinese Han population. AB - Killer cell immunoglobulin-like receptors (KIRs) on natural killer cells recognize groups of HLA class I alleles. Seventeen KIR genes have been identified at present, and two kinds of KIR haplotypes (group A and B) have been described based on their gene contents. Immunogenetic analysis of different ethnic populations shows significant differences in terms of the distribution of group A and B haplotypes. Here, genomic DNA from 104 healthy unrelated Chinese Han individuals was typed for the presence or absence of KIR genes. All 17 KIR genes were observed in the population, and framework genes 3DL3, 3DP1, 2DL4, and 3DL2 were present in all individuals. Twenty-six different genotypes were found, four of which could not be assigned to haplotypes according to the model of Hsu et al. (J Immunol 2002: 169: 5118). Group A haplotypes outnumbered group B haplotypes in frequency by approximately 3:1, with individuals having two group A haplotypes accounting for 58.7%. Analysis indicated that some pairs of KIR genes showed remarkable linkage disequilibrium. Our data demonstrated that the Chinese Han population is distinct in KIR gene frequencies and putative KIR haplotypes in comparison to some other populations. PMID- 15896205 TI - Identification of a novel HLA-A*0278 allele in a Chinese family. AB - A novel human leukocyte antigen-A (HLA-A) allele, A*0278, has been identified in a Chinese family using DNA-based typing and molecular cloning methods. The alleles A*0278 differs from its closest matching HLA sequence of A*0256 by a silent substitution at 102 A > C and by two replacement substitutions, 98T > A and 292 C > G in exon 2, resulting in a change of codon 33 from Phe (TTC) to Tyr (TAC) and codon 98 from His (CAC) to Asp (GAC). Serology study revealed that A*0278 is associated with HLA-A2 broad specificity. A polymerase chain reaction sequence-specific primers-based assay was developed to identify A*0278. Family study indicated that the propositus inhered his father's HLA haplotype A*0278, B*35, DRB1*15. No further individuals of A*0278 were found in 5000 Chinese bone marrow donor volunteers. PMID- 15896206 TI - A novel A*24 allele, A*2442, was detected through routine bone marrow donor screening. AB - In this article, we report the identification of a new HLA-A allele found in a DNA sample which was part of the routine bone marrow donor typing performed in our laboratory. This novel allele officially designated as A*2442 was found in a sample from a female Caucasoid donor (Franken, Bavaria, Germany; lab code 142654) and differs from the closest related allele A*2408 by two nucleotide exchanges. In position 81, the A (A*2408) is changed to 81 C in the novel allele A*2442, resulting in an amino acid substitution at codon 27, glutamine (A*2408) is replaced by histidine ((31)Gln-->(31)His). In position 292, the G (A*2408) is changed to C also resulting in an amino acid replacement, the codon 98 asparagine is mutated to histidine ((98)Asn-->(98)His) in the new A*2442 allele. The second allele was determined to be A*0301. Further typing of this sample is B*1501 B*4001. PMID- 15896207 TI - HLA-B*5701 typing by sequence-specific amplification: validation and comparison with sequence-based typing. AB - Susceptibility to abacavir hypersensitivity (ABC HSR) is strongly associated with alleles carried on the 57.1 ancestral haplotype including HLA-B*5701 and Hsp70 Hom M493T. In one study, prospective testing for HLA-B*5701 and exclusion of individuals carrying this allele, from receiving abacavir, substantially lowered the incidence of ABC HSR to 0% (95% confidence interval 0-0.075%). The presence of HLA-B*5701 is usually detected by standard serological tests and by molecular genetic methods such as sequence-based typing (SBT). While the former test cannot discriminate between HLA-B57 subtypes, the expensive SBT may not be readily available in all laboratories. Hence, an alternate method was developed to detect HLA-B*5701 using allele and group-specific polymerase chain reaction-sequence specific primers (PCR-SSP) typing. This PCR-SSP-typing method positively amplified all HLA-B*5701 alleles in concordance with their SBT-assigned typing. This multiplexed SSP assay was able to distinguish between HLA-B*5701 (n = 10) and closely related HLA-B57 alleles B*5702 (n = 2), -B*5703 (n = 1), -B*5704 (n = 1) alleles and non-HLA-B*57 alleles (n = 61). In conclusion, this method of HLA B*5701 detection is a rapid and accurate typing method with high specificity, sensitivity and reproducibility. PMID- 15896208 TI - Characterization of a novel HLA-Cw*02 variant, Cw*0208, in a Caucasian individual. AB - We describe an additional HLA-Cw*02 variant, HLA-Cw*0208, which has been identified in a renal transplant recipient of Caucasian origin (Italy). After performing preliminary serological typing, we analyzed exons 2 and 3 of the HLA-C locus polymorphism by cloning the amplified DNA and using a sequence-based typing method. The new allele differs from Cw*020202 by one nucleotide substitution at nucleotide 61 (G-->A) of exon 2, which translates to a difference of one amino acid at residue 21 (His-->Arg) of the HLA-C heavy chain. We propose that Cw*0208 was generated by a random point mutation in codon 21 from the Cw*020202 allele, or through gene conversion of Cw*020202 with another allele, probably the Cw*1205 and Cw*1602 alleles. PMID- 15896209 TI - A novel HLA-Cw*16 allele, Cw*160102, identified by cloning and sequencing. PMID- 15896210 TI - Identification of a new HLA-DRB1 allele, HLA-DRB1*1212, and confirmation of HLA B*1586*. AB - A novel allele, HLA-DRB1*1212, was identified by sequence-based typing. DRB1*1212 was identical to DRB*120101 at exon 2 except for a single nucleotide at position 199 from A to C, leading to amino acidic substitution from Ile to Leu at codon 67. We also confirmed the HLA-B*1586 in another Chinese person. PMID- 15896211 TI - Nomenclature for factors of the HLA system, update January 2005. PMID- 15896212 TI - Nomenclature for factors of the HLA system, update February 2005. PMID- 15896213 TI - Nomenclature for factors of the HLA system, update March 2005. PMID- 15896214 TI - The management of bipolar disorder in primary care: a review of existing and emerging therapies. AB - Recent evidence suggests that the prevalence of bipolar disorder is as much as fivefold higher than previously believed, and may amount to nearly 5% of the population, making it almost as common as unipolar major depression. It is, therefore, not unrealistic to assume that primary care or family physicians will frequently encounter bipolar patients in their practice. Such patients may present with a depressive episode, for a variety of medical reasons, for longer term maintenance after stabilization, and even with an acute manic episode. Whatever the reason, a working knowledge of current trends in the acute and longer-term management of bipolar disorder would be helpful to the primary care physician. In addition, an understanding of important side-effects and drug interactions that occur with drugs used to treat bipolar disorder, which may be encountered in the medical setting, are paramount. This paper will attempt to review existing and emerging therapies in bipolar disorder, as well as their common drug interactions and side-effects. PMID- 15896215 TI - Geropsychiatric consultation in a general hospital in Taiwan. AB - The aim of this study was to characterize clinically significant issues in a psychiatric consultation service for geriatric inpatients in a general hospital in Taiwan. This was a case-control study. During a 5-month period, 100 geriatric (age > or =65 years) inpatients consecutively referred for consultation-liaison psychiatric service from non-psychiatric departments formed the study group. Another 100 medical inpatients, also referred for consultation-liaison to the psychiatric service, but aged 17-50, formed the control (non-geriatric) group. The diagnosis, demography, reason for referral, symptomatology, and other clinical characteristics were determined by consensus between two psychiatrists. Psychiatric diagnosis was made according to criteria in the 4th edition of the Diagnostic and Statistical Manual of Mental Disorders. The geropsychiatric consultation rate was 0.9%. Geriatric patients constituted 20.1% of all psychiatric referrals. Common reasons for referral of geriatric inpatients were confusion (32%), depression (17%), disturbing behaviors (14%), and psychosis (14%). The most common psychiatric disorder among geriatric patients was an organic mental disorder (79%), followed by a depressive disorder (13%). More geriatric patients suffered from cancers and cerebrovascular diseases than non geriatric patients. The geriatric group was more likely to have multiple physical illnesses. Organic mental disorder and depressive disorders are the most common psychiatric diagnoses in the geropsychiatric consultation service of the authors. In the authors' experience, both psychotropic medication treatment and psychosocial intervention are important in geropsychiatric consultation. PMID- 15896216 TI - Efficacy of 3-year psychiatric daycare treatment in patients with schizophrenia. AB - The present study investigated the efficacy of a 3-year psychiatric daycare (DC) program with regard to psychiatric symptoms and difficulties with daily living experienced by patients with schizophrenia. The subjects were 28 patients who met the diagnostic criteria from the 4th edition of the Diagnostic and Statistical Manual for schizophrenia and continued DC treatment for 3 years. The present study assessed participants at two points: at the start of DC and after 3 years, by evaluating socioeconomic factors and Brief Psychiatric Rating Scale (BPRS) scores. In addition, in order to measure difficulties with daily living, the Life Assessment Scale for the Mentally Ill (LASMI) and the Etoh Daycare Assessment Scale (ETODAS) developed at the Etoh Hospital, were used. Results indicated that no significant changes in socioeconomic factors or BPRS scores occurred during the 3-year period of DC treatment. However, over this period, mean scores (+/- standard deviation [SD]) for LASMI subcategories decreased from 1.6 +/- 0.8 points to 0.9 +/- 0.7 for daily living and from 1.7 +/- 0.8 to 1.2 +/- 0.7 for interpersonal relations, indicating significant improvement (P < 0.05). In addition, mean scores (+/-SD) for the ETODAS subcategories increased from 3.4 +/- 0.8 to 4.1 +/- 0.8 for expressiveness, from 3.3 +/- 0.9 to 4.0 +/- 0.8 for communication, from 3.1 +/- 0.6 to 3.6 +/- 1.0 for initiative within a group, and from 3.4 +/- 0.5 to 3.8 +/- 0.7 for cooperation in work activities, indicating significant improvement (P < 0.05). The present study suggests that DC can enable patients with schizophrenia to maintain their condition without worsening the psychiatric symptoms, and to improve their daily living skills, social skills in human relations, and work skills. PMID- 15896217 TI - Delirium in inpatients with respiratory diseases. AB - The features of delirium in patients being hospitalized due to respiratory diseases were investigated. From the inpatients in the respiratory medical ward of Tokyo Metropolitan Hiroo General Hospital over the course of 1 year, the patients who had delirium were diagnosed by a semistructured interview. The total number of subjects was 454, and patients with delirium were 43. Various clinical factors were compared between the delirium group and non-delirium group. In the delirium group, there were many elderly patients of 70 years or older. Moreover, there were many patients who had a chronic respiratory disease, patients in which the respiratory diseases were mutually complicated, and patients in whom other diseases combined with the respiratory disease in the delirium group. There were also many patients in the Intensive Care Unit (ICU), and patients with an endotracheal intubation or extubation. Based on the results of a multiple logistic regression analysis, for age, ICU accommodation, and endotracheal intubation, the value of the delirium group was more significant than that of the non-delirium group. In half of the patients from the delirium group, delirium developed within 1 week after hospitalization. In the patients who died in the hospital, however, delirium often developed days after they had been hospitalized. It was suggested that the later developed delirium had a relation to the prognosis. PMID- 15896218 TI - Relation of surgery, tumor site, and age group to the loss of reality testing in Japanese patients with malignant tumors: a study of a hospital-based sample with a consultation-liaison service. AB - The relation between surgery and the loss of reality testing (LRT) in Japanese patients with malignant tumors were examined, taking into account the influence of the tumor sites and age groups. The patients were comprised of 277 men and 225 women with malignant tumors in Kyushu University Hospital, Fukuoka, Japan, who underwent a check-up at the Department of Neuropsychiatry for the first time using the consultation-liaison system. Those with known schizophrenia, dementia, mental retardation, and paranoid or schizoid (schizotypal) personality disorder were excluded. Surgery was statistically significantly associated with LRT in elderly men (> or =65 years of age), and in men with malignant tumors of the digestive organs (odds ratio [OR], 9.7; 95% confidence interval [95% CI], 3.2 29.3). Even after adjusting for tumor site and age, surgery was statistically significantly associated with LRT in men (OR, 2.6; 95% CI, 1.4-4.6) and nearly significantly associated in women (OR, 1.8; 95% CI, 0.9-3.6). There were no material associations between surgery and LRT in patients with malignant tumors of sex-specific organs or the head and neck area. The present study showed a positive relationship between surgery and LRT in Japanese men and women with malignant tumors. The association was stronger in elderly patients. As for tumor site, surgery was most strongly associated with an increased risk of LRT in patients with malignant tumors of the digestive organs. PMID- 15896219 TI - Gender differences in association of plasma adiponectin with obesity reflect resultant insulin resistance in non-diabetic Japanese patients with schizophrenia. AB - Adipose tissues poorly produce adiponectin in the population with increased body fat mass and diabetes mellitus. It was investigated whether hypoadiponectinemia is associated with obesity and insulin resistance in patients with chronically medicated schizophrenia. A cross-sectional study was designed for 73 non-diabetic Japanese patients with schizophrenia. The patients aged <70 years with body mass index (BMI) > or =18.5 were selected. Anthropometrics and blood parameters including fat-derived cytokines were measured, and then the BMI and homeostasis model assessment-insulin resistance (HOMA-IR) was calculated. The variables were compared between the non-obesity (BMI, 18.5-24.9) and obesity (> or = 25.0) groups, and between genders. Plasma adiponectin negatively correlated with BMI (r = -0.554, P < 0.0003) and HOMA-IR (r = -0.380, P = 0.007) in men, but not in women. The obesity group in men, as compared with the non-obesity group, showed significantly lower plasma adiponectin (P = 0.008) and higher HOMA-IR (P < 0.05), but not in women. Plasma leptin showed a significant positive correlation with BMI (r = 0.604, P < 0.0001 in men; r = 0.763, P < 0.0001 in women) and HOMA-IR (r = 0.618, P < 0.0001 in men; r = 0.679, P < 0.0001 in women). The mean plasma leptin in the obesity group was significantly higher than that in the non-obesity group (P < 0.01 in men; P < 0.01 in women). In contrast to plasma leptin, plasma adiponectin showed gender difference in relation to BMI and HOMA-IR. PMID- 15896220 TI - Behavioral and psychological symptoms of dementia characteristic of mild Alzheimer patients. AB - In order to clarify the characteristics of Behavioral and Psychological Symptoms of Dementia (BPSD) in patients with mild Alzheimer's disease (AD), BPSD among the severities of Clinical Dementia Rating (CDR) in 74 patients with AD were compared using the Neuropsychiatric inventory (NPI). The result, when compared between mild (CDR = 0.5, 1) and moderate or severe (CDR = 2, 3) AD, was a significant difference in frequency of euphoria, disinhibition and aberrant motor behavior, but no significant difference was found in frequency of delusions, hallucinations, agitation, dysphoria, anxiety, apathy and irritability. In addition, a significant difference was found in the mean scores of the composite score for euphoria, apathy, disinhibition and aberrant motor behavior, but no significant difference was found in the mean scores of the composite score for delusions, hallucinations, agitation, dysphoria, anxiety and irritability. That is, the mild AD groups (CDR 0.5 or 1) had delusions, hallucinations, agitation, dysphoria, anxiety, apathy and irritability as frequently as the moderate or severe AD groups (CDR 2 or 3), and had the equivalent level of composite scores to the moderate or severe AD groups (CDR 2 or 3) in delusion, hallucination, agitation, dysphoria, anxiety and irritability. Therefore, it was supposed that psychotic symptoms (delusion, hallucination) and emotional symptoms (agitation, dysphoria, anxiety, irritability) are important BPSD in patients with mild AD as well as those with moderate or severe AD, and there are needs for health, welfare and medical services for these symptoms. PMID- 15896221 TI - Regional cerebral blood flow deficits in mild Alzheimer's disease using high resolution single photon emission computerized tomography. AB - In spite of its wide availability, single photon emission computerized tomography (SPECT) scanning is uncommonly used in the assessment of Alzheimer's disease (AD) and related dementias. In light of recent advances in scanning protocols and image analysis, SPECT needs to be re-examined as a tool in the diagnosis of dementia. A total of 18 subjects with early AD and 10 healthy elderly control subjects were examined with high resolution SPECT during the performance of a simple word discrimination task. SPECT images were coregistered with individual magnetic resonance imaging scans, allowing delineation of predetermined neuroanatomical Regions of Interest (ROI). There was a gradation of regional cerebral blood flow (rCBF) values in both groups, with the lowest values being in the hippocampus and the highest in the striatum, thalamus and cerebellum. Compared to healthy controls, AD subjects demonstrated lower relative rCBF in parietal and prefrontal cortices. Analysis of individual ROI demonstrated bilateral reduction of rCBF in prefrontal poles, posterior temporal and anterior parietal cortex, and unilateral reduction of rCBF in left dorsolateral prefrontal cortex, right posterior parietal cortex and the left cingulate body. There were no significant differences for hippocampal, occipital or basal ganglia rCBF. Discriminant function analysis indicated that rCBF in the prefrontal polar regions achieved the best classification of cases. SPECT has utility in the diagnostic assessment of AD if standardized and semiquantitative techniques are used. PMID- 15896222 TI - Obsessive-compulsive symptoms in schizophrenia: prevalance and clinical correlates. AB - Obsessive-compulsive symptoms (OCS) have been observed in a substantial proportion of schizophrenic patients. In this study, the rate of occurrence of OCS and obsessive-compulsive disorder (OCD) in schizophrenic patients, and also the interrelationship between OCS and schizophrenic symptoms and depressive symptoms were assessed. A total of 100 subjects with a diagnosis of schizophrenia from the 4th edition of the Diagnostic and Statistical Manual (DSM-IV) were evaluated by the structured and clinical interview for axis-1 DSM-IV disorders patient edition (SCID-P), the Positive and Negative Syndrome Scale (PANSS), Yale Brown Obsessive-Compulsive Scale (Y-BOCS), and the Calgary Depression Rating Scale for Schizophrenia. The prevalance of OCS in individuals meeting criteria for schizophrenia was 64%. A total of 30 of these patients (Y-BOCS total score > or =7) also met the DSM-IV criteria for OCD. The total score on Y-BOCS was significantly correlated with total score on PANSS, Positive-PANSS score, General PANSS score and total score on Calgary Depression Rating Scale for Schizophrenia. OCS and OCD relatively frequent in schizophrenic patients and OCS are significantly correlated with the severity of psychosis, positive symptoms, and depressive symptoms in schizophrenic patients. These findings provide further evidence for the importance of OCS in schizophrenia. PMID- 15896223 TI - Suicide patterns and characteristics in Akita, Japan. AB - Akita Prefecture currently has the highest rate of suicide in Japan. Given this alarming statistic, investigation of the underlying causes of suicide and identification of strategies for suicide prevention are imperative. Members of the Akita Prefectural Medical Association (APMA) see most of the individuals who commit suicides in Akita Prefecture, so data from the APMA would prove advantageous in any investigation of suicides. In this study, members of the APMA who had attended to individuals who had committed suicide were asked to complete a questionnaire about the case to determine the factors underlying suicide in Akita Prefecture. From 1 July 2001 to 30 June 2002, a total of 138 cases (102 males, 36 females) of suicide were reported. Most suicide cases were of 50-69 year olds. Many cases involved relatively lethal methods (such as hanging). Most suicides were performed at home and at a time when the rest of the family was asleep or absent. The most common complaint appeared to be economic-related problems. Depressive disorder was the most common psychiatric disorder, and many cases displayed high depressive trait scores. The present results do not exclude the possibility that economic-related problems are playing a major role in recent increases in suicide numbers. However, strategies for dealing with depression as well as economic-related problems are considered important. PMID- 15896224 TI - Gait disturbances in patients with schizophrenia and adaptation to treadmill walking. AB - This study evaluated the gait patterns of schizophrenic patients at free gait and at three fixed velocities on a treadmill. The effects of illness and antipsychotic treatment on gait parameters and on adaptation to treadmill walking were compared. Gait parameters of 14 drug-naive schizophrenic patients, 14 patients treated with conventional antipsychotics, 14 patients treated with olanzapine, as well as 14 matched controls were assessed on a walkway and on a treadmill at three different velocities (very slow, intermediately slow, and comfortable) using an ultrasonic movement analysis system. At free gait, all patients showed a significantly decreased gait velocity, predominantly due to a shorter stride length, when compared to the controls, with the most striking difference observed between the patients treated with conventional neuroleptics and the controls (ANOVA, P < or = 0.001). Cadence (steps per second) did not differ between the investigated groups. When gait was evaluated on the treadmill, differences in stride length and cadence were significant only at the very slow treadmill velocity (ANOVA, P < or = 0.05). In all patient groups, mean stride length was decreased and cadence compensationally increased. Significant differences between the patient groups were no longer detectable. With increasing treadmill velocities, gait parameters of all patient groups normalized. The results show that, like in patients with Parkinson's Disease, impaired gait parameters can also be normalized in schizophrenic patients by external stimulation via treadmill walking. PMID- 15896225 TI - Drug preferences in illicit drug abusers with a childhood tendency of attention deficit/hyperactivity disorder: a study using the Wender Utah Rating Scale in a Japanese prison. AB - The purpose of this study is to clarify the relationship between childhood tendencies of attention deficit/hyperactivity disorder (AD/HD) and illicit drug abuse in Japanese prisoners, and to clarify whether drug abusers with AD/HD prefer methamphetamine (MAP) more than other illicit drugs. The Japanese version of the Wender Utah Rating Scale (WURS), which is a self-reporting instrument to retrospectively identify childhood tendencies of AD/HD tendencies, was carried given to 413 prisoners without a drug addiction and 282 prisoners with a drug addiction (192, MAP; 53, toluene; and 37, cannabis). WURS scores were compared between prisoners with and without a drug addiction, and between MAP, toluene, and cannabis abusers. Consequently, prisoners with a drug addiction showed significantly higher WURS scores than those without the addiction (P < 0.001). Toluene abusers showed significantly higher WURS scores than cannabis abusers (P < 0.001), and included a higher proportion with scores over cut-off than MAP or cannabis abusers (P = 0.005). In conclusion, a close relationship existed between illicit drug abuse and childhood AD/HD tendencies. Drug-abusing prisoners with AD/HD tendencies were not prone to choose MAP over other illicit drugs. PMID- 15896226 TI - Characteristics of self-cutters among male inmates: association with bulimia and dissociation. AB - It was examined whether bulimia and dissociation are common in male self-cutters, as has been found in female self-cutters. The subjects were 796 male inmates of a juvenile prison. A self-reporting questionnaire was used to assess self-cutting, histories of psychoactive substance use, problem behaviors, and traumatic life events in the subjects. The Adolescent Dissociative Experience Scale and the Bulimia Investigatory Test of Edinburgh were also used. Subjects were divided into two groups: self-cutting and non-cutting. Questionnaire responses and dissociation and bulimia assessments were compared between the groups. Self cutters began smoking (P < 0.001) and drinking (P < 0.001) earlier, and more frequently used illicit psychoactive drugs (P < 0.001), experienced childhood physical abuse (P < 0.001), and reported suicide attempts (P < 0.001), suicidal ideation (P < 0.001), and outward violence toward a person (P < 0.001) or object (P < 0.001) than non-cutters. Self-cutters also scored significantly higher on the bulimia (P < 0.001) and dissociation tests (P < 0.001). Logistic regression analysis demonstrated that suicide attempt (odds ratio, 4.311) and suicidal ideation (odds ratio, 2.336) could discriminate between male inmates with and without self-cutting. Male self-cutters showed 'multi-impulsive bulimic' tendencies resembling those of female self-cutters, although to a lesser extent. Clinical features of male as opposed to female self-cutters were influenced by gender differences. PMID- 15896227 TI - Factors associated with suicidal ideation in an elderly urban Japanese population: a community-based, cross-sectional study. AB - The objective of the present study was to evaluate the association between suicidal ideation and potentially related factors in an elderly urban Japanese population. This was a community-based, cross-sectional study. Urban community residents aged 70 years or more were interviewed regarding suicidal ideation and sociodemographic and health-related variables. Subjects with depressive symptoms underwent further evaluation by psychiatrists using criteria of the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. Associations were evaluated using univariate and multivariate logistic regression analyses. Of the 1145 eligible participants, 52 (4.5%) reported thoughts of suicide. Of 143 subjects with depressive symptoms (Geriatric Depression Scale, 14+), 22 (15.4%) reported suicidal ideation over a 2-week period. After controlling for depressive symptoms, lack of social support and impaired instrumental activities of daily living were significantly associated with thoughts of suicide. After controlling for the potentially associated factors detected in the univariate analysis, depressive symptoms were strongly associated with thoughts of suicide. In the elderly with depressive symptoms, mental disorders, including depressive and alcohol-related disorders, were significantly associated with suicidal ideation over a 2-week period. In the urban community setting, screening for lack of social support, impaired instrumental activities of daily living, and depressive symptoms, followed by diagnostic evaluation for mental disorders, particularly for depressive syndromes and alcohol-related disorders, may provide a practical and effective means of identifying elders at high risk of suicide. PMID- 15896228 TI - Community-based suicide prevention through group activity for the elderly successfully reduced the high suicide rate for females. AB - The lack of social support is the most common risk factor for late-life suicide. Few previous community-based interventions against the lack of social relationships reduced suicide. This study aims to evaluate outcomes of a community-based prevention program against suicide amongst the elderly in rural Japan. During 1995-2002, the program based on population strategy including group activity, psychoeducation and self-assessment of depression but no screening for depression, was implemented for elderly residents in Yuri town, Japan (5-year average population 6817; 5-year average suicide rate [> or =65 years old] 291.4/10(5)). Changes in the relative risk of suicide for individuals (> or =65 years old) before and after the 8-year implementation were estimated by the incidence rate ratio (IRR), using a quasi-experimental design with a neighboring reference, Chokai town, Japan (5-year average population 8136; 5-year average suicide rate [> or =65 years old] 216.5/10(5)). The risk of elderly females in Yuri completing suicide was reduced by 76% (age-adjusted IRR, 0.24; 95% CI, 0.10 0.58), while there was no change in the risks for Yuri's elderly males and both Chokai's elderly males and females. General loglinear analysis estimated a ratio of the female IRR in Yuri to that in its Akita prefecture of 0.35 (95% CI, 0.14 0.84), showing that the reduction of the risk in the intervention area was greater than a historical trend. A community-based suicide prevention through a group intervention designed to increase knowledge and to cultivate social relationships would be effective for elderly females but not males. PMID- 15896229 TI - Lack of an association between 5-HT receptor gene polymorphisms and suicide victims. AB - An association between serotonergic dysfunction in the brain and suicidal behavior has previously been suggested. The high affinity of some antipsychotic and antidepressant drugs to serotonin 6 (5-HT6) receptors, and the predominant localization of 5-HT6 receptors in some limbic regions, suggest that 5-HT6 receptors play a role in the pathogenesis of suicide. The objective of the present study was to examine the association between suicide victims and two polymorphisms of the 5-HT6 receptor gene: a biallelic polymorphism (267C/T) in exon 1 and a trinucleotide repeat polymorphism ([GCC](2/3)) in the 5'-upstream region of the gene. The two polymorphisms were genotyped in 163 suicide victims and 166 controls, and the distribution of genotype and allele frequencies between the two groups were compared. Haplotype frequencies of these two polymorphisms were estimated from genotypic data by the maximum-likelihood method. In both polymorphisms, there were no significant differences in genotype or allele frequencies between the suicide victims and the controls. Moreover, there were no significant differences in the haplotype distributions of these polymorphisms between the two groups. These findings suggest that it is unlikely that the 5-HT6 receptor gene is involved in the susceptibility to suicide. PMID- 15896230 TI - Prevalence of restless legs syndrome and associated factors in the Korean adult population: the Korean Health and Genome Study. AB - The present study was purposed to identify the prevalence of restless legs syndrome (RLS) and its associated factors in the Korean adult population. Among a total of 9939 participants derived from the Korean Health and Genome Study, 12.1% of subjects (men, 8.5%; women, 15.4%) suffered from RLS. Factors independently related with RLS were older age and frequent fatigue in both men and women. PMID- 15896231 TI - Transcranial magnetic stimulation for the deficit syndrome of schizophrenia: a pilot investigation. AB - In an open study, four subjects with a stable deficit syndrome of schizophrenia received high frequency repetitive transcranial magnetic stimulation (15 Hz at 90% of motor threshold, 1800 pulses each session, daily for 20 sessions over 4 weeks) over the left dorsolateral prefrontal cortex. Subjects showed a significant reduction in negative symptoms and improvement in function, with no change in positive symptoms. This improvement was maintained at the 1 month follow up. Repetitive transcranial magnetic stimulation as a treatment of the deficit syndrome of schizophrenia is feasible, safe and may be beneficial. A systematic study in randomized control trials would be appropriate. PMID- 15896232 TI - Effects of intraperitoneally injected lithium, imipramine and diazepam on nitrate levels in rat amygdala. AB - Nitric oxide (NO) has been studied in relation to the etiologies of various neurologic and psychiatric diseases. However, little is known about whether clinically available psychotropic drugs affect the NO system in the brain. Using an in vivo brain microdialysis method, the effects of intraperitoneally administered lithium, imipramine and diazepam on levels of , a marker of in vivo NO production, were investigated in the rat amygdala. Lithium significantly reduced, while imipramine raised, levels as compared with controls. These observations suggest that lithium and imipramine induce opposite effects on NO related systems in the brain. PMID- 15896233 TI - Disappearance of specific phobia after removal of ventricular glioblastoma. PMID- 15896234 TI - The effect of aromatherapy massage on mild depression: a pilot study. PMID- 15896235 TI - Patient valuation of pharmacist services for self care with OTC medications. AB - BACKGROUND AND OBJECTIVES: The objectives of this study were to determine whether patients would be willing to pay for pharmacist self-care services on proper use of over-the-counter medications. In addition, we examined whether patients' willingness to pay was associated with community pharmacy setting and patients' socio-economic factors. METHODS: A self-administered questionnaire was handed out to patrons of community pharmacies in Arkansas, USA. The questionnaire contained question items related to patients' willingness to pay, community pharmacy setting, prescription drug insurance and socio-economic information. Patients' willingness to pay was measured using the checklist method. RESULTS AND DISCUSSION: A total of 262 questionnaires were completed. More than twice as many patients (51%) are now willing to pay for pharmacist services for patient self care than a decade ago. They were willing to pay about US5 dollars for a 5-min consultation. Willingness to pay was significantly associated with community pharmacy setting (chi-square, P = 0.030). Grocery/chain pharmacy patrons were more willing to pay than independent pharmacy patrons. CONCLUSION: This increased patient willingness to pay, along with growing self-care markets, provides pharmacists with opportunities to develop self-care clinics or services. PMID- 15896236 TI - Reduced oral itraconazole bioavailability by antacid suspension. AB - AIMS: To investigate the effects of antacid suspension on oral absorption of itraconazole. METHODS: A randomized, open-labelled, two-period, crossover study with a 1-week washout period was conducted in 12 healthy Thai male volunteers. The participants were allocated in either treatment A or B in the first period. In treatment A, the volunteers were orally administered with 200 mg of itraconazole alone. In treatment B, the volunteers were administered orally with 200 mg of itraconazole co-administered with antacid suspension. Serial serum samples were collected over the period of 24 h and subsequently analysed by using a validated high-pressure liquid chromatographic method with ultraviolet detection. Pharmacokinetic parameters were determined by non-compartmental analysis. RESULTS: Time to reach maximal concentration (Tmax), maximal concentration (Cmax) and area under the curve (AUC0-infinity) were markedly decreased in antacid-treated group. Tmax for treatment A was 3.0 +/- 0.4 and 5.1 +/- 2.7 h for treatment B. Cmax and AUC0-infinity of treatments A and B were 146.3 +/- 70.5 vs. 43.6 +/- 16.9 (ng/mL) and 1928.5 +/- 1114.6 vs. 654.8 +/- 452.2 (ng x h/mL) respectively. 90% Confidence interval (90% CI) of Cmax and AUC0 -infinity were 24.1-42.1 and 16.2-65.9 respectively. CONCLUSIONS: Rate and extent of itraconazole oral absorption were markedly decreased by concurrent use of antacid suspension. Hence, co-administration of itraconazole and antacid suspension should be avoided. PMID- 15896237 TI - Administration of enoxaparin by continuous infusion in a naturalistic setting: analysis of renal function and safety. AB - STUDY OBJECTIVE: To describe the clinical use and safety of continuous infusion (CI) enoxaparin in a naturalistic setting and to evaluate the influence of renal function on enoxaparin elimination. DESIGN: Retrospective medical record review. SETTING: 1000-bed tertiary care teaching centre. PATIENTS: Hospitalized patients that received enoxaparin by CI during a 2-year period. INTERVENTIONS: None. MEASUREMENTS: Specific details of dosage and monitoring were collected. Adverse drug reactions (ADR) were recorded. Creatinine clearance (CrCl) was calculated using Cockroft and Gault and Brater equations. A population pharmacokinetic analysis was performed using the non-linear mixed effect model (NONMEM). For patients located in the intensive care unit (ICU) and ward, POSTHOC pharmacokinetic parameter estimates were evaluated using the Wilcoxon rank-sum. Pearson correlation coefficient was calculated to determine the association between renal function and anti-Xa clearance. MAIN RESULTS: Sixty-seven patients received enoxaparin by CI of which 61.2% were in the ward and 38.8% in the ICU. The average initial rate and duration of infusion were 5.2 mg/h and 5.6 days, respectively. The number of anti-Xa concentration measurements averaged five per patient. Nine patients experienced an ADR. The most frequent ADR was gastrointestinal bleeding (n = 4). Among the 67 patients, 48 had available anti Xa concentrations and were included in the NONMEM model. The anti-Xa CL and volume of distribution for ICU and ward patients averaged 0.64 +/- 0.34 L/h, 10.6 +/- 1.55 L and 1.01 +/- 0.39 L/h, 9.08 +/- 1.17 L, respectively. CrCl was not a significant covariate when included in the NONMEM model, and the association between CrCl and anti-Xa clearance was not significant (R2 = 0.0005; P = 0.8916). CONCLUSIONS: This study is the first to report the use and safety of prolonged CI enoxaparin. Pharmacokinetic parameters of enoxaparin differ in ICU vs. ward patients. Overall, we found the safety of CI to be comparable to subcutaneous administration. Also, we found no effect of renal function on enoxaparin elimination. PMID- 15896238 TI - User-related pharmaceutical care problems and factors affecting them: the importance of clinical relevance. AB - BACKGROUND AND OBJECTIVES: Many studies determined the number and nature of user related Pharmaceutical Care Problems (PCP) and factors affecting them, but none considered the inclusion of clinical relevance. The aims of this study are (i) to investigate the type, number and clinical relevance of user-related PCP self reported by home dwelling elderly on polypharmacy and (ii) to develop a risk model for detecting elderly drug-users at risk of user-related PCP. METHODS: The study was a cross-sectional study conducted among 286 home dwelling elderly on polypharmacy (> or =75 years, > or =4 medicines) in the Netherlands. The user related PCP found were divided into problem categories and subsequently a pharmacist and a general practitioner classified the problems into those with low and those with (potential) clinical relevance. Factors possibly associated with PCP (both for all and relevant problems) were identified, and subsequently tested in multivariate models using logistic regression. RESULTS: Three hundred and ninety-eight user-related PCP were observed in 189 patients (66% of all participants). After classification of user-related PCP only 26% appeared to be of clinical relevance (26% of all participants). When including clinical relevance a shift in predominantly present problem categories is observed. Furthermore, the risk model for problems with clinical relevance contains more factors than the model which considered all problems. Factors associated with clinically relevant PCP are emotional or physical problems interfering with social life, communication skills (vision and hearing), using tablets that have to be divided, using inhaled medicines, and the number of medicines used. This risk-model has a specificity of 92% and a sensitivity of 32%. CONCLUSIONS: Although user-related PCP were seen in about two-thirds of the participants, in only one out of four participants was the PCP considered to be of clinical relevance. With inclusion of clinical relevance, other problem categories become more dominant. A specific risk model is designed to select elderly patients that are most likely to have PCP in need of more urgent intervention. Unfortunately higher specificity is accompanied by low sensitivity in the present model. PMID- 15896239 TI - Advice-giving on self-medication: perspectives of community pharmacists and consumers in Singapore. AB - BACKGROUND: In Singapore, community pharmacists provide an advice-giving service to consumers who seek self-medication for minor ailments management. This service has not been studied formally from the perspectives of pharmacists and consumers. OBJECTIVES: The study aimed to identify (i) the approach taken by pharmacists in providing advice for self-medication and (ii) consumers' behaviour in self treatment and their perception of the advice-giving role of the community pharmacist. METHOD: The pharmacists and consumers were surveyed independently using two structured questionnaires. RESULTS AND DISCUSSION: All community pharmacists who participated in the survey were confident in providing advice on self-medication. However, none of them recorded the consultations and only 17.5% of them had documented their general physician referrals. Most consumers (66.3%) would self-medicate and only consult a professional when the desired outcome was not achieved. Less than 10% of consumers would approach the pharmacists as the first option for advice. More than half of the pharmacists felt that the advice they rendered deserved a fee whereas only 28.4% of the consumers were willing to pay. Both parties thought the fee should not be more than S5 dollars (US3 dollars). CONCLUSION: Generally, there is congruence in the perspectives on self medication between the advice-giving pharmacist and the consumer. The consumers still lack awareness that pharmacists can help them to self-medicate more safely and effectively. Therefore, more effort in public education is warranted. The current state of poor documentation of the advisory function of community pharmacists should be improved. PMID- 15896240 TI - Clinical factors associated with hyperkalemia in patients with congestive heart failure. AB - BACKGROUND: Patients with congestive heart failure (CHF) are at risk for hyperkalemia because of coexisting comorbidities and use of multiple medications that impair potassium (K) excretion such as angiotensin converting enzyme (ACE) inhibitors. OBJECTIVE: To identify clinical factors associated with hyperkalemia on initial presentation in patients hospitalized for CHF. DESIGN: A case-control study. SETTING: Two university-affiliated tertiary-care hospitals. SUBJECTS: Using ICD-9 code for CHF, CHF admissions with hyperkalemia on presentation (cases) were identified from a population of 938 non-dialysis-dependent CHF patients. CHF admissions with normokalemia on presentation were used as controls. Hyperkalemia was defined as serum K > or = 5.6 mmol/L, and normokalemia as serum K > or = 3.5 and < or =5.5. METHODS: Data were collected on demographic characteristics, clinical variables, comorbidity and medication use. Factors associated with hyperkalemia on initial presentation were examined. RESULTS: Mean age did not differ between cases [76 years, standard deviation (SD) = 12] and controls (75 years, SD = 12) (P = 0.824). Mean potassium levels for cases and controls were 6.2 mmol/L (range 5.6 to 8.2) and 4.3 mmol/L respectively (P < 0.001). On multivariate analysis, diabetes mellitus [odds ratio (OR) = 2.42, 95% confidence interval (CI) = 1.04-5.59], creatinine clearance <40 mL/min (OR = 8.36, CI = 2.73-25.56), use of spironolactone (OR = 4.18, CI = 1.27-13.79), and use of ACE inhibitors (OR = 2.55, CI = 1.06-6.13) were independently associated with hyperkalemia. CONCLUSIONS: In CHF patients, hyperkalemia on presentation is independently associated with diabetes, creatinine clearance <40 mL/min, use of spironolactone, and use of ACE inhibitors. Recommendations for use of spironolactone and ACE inhibitors in CHF, and the intensity of serum K monitoring need to be clarified to account for patients at higher risk for hyperkalemia. PMID- 15896241 TI - Relationship of P450 2C9 genetic polymorphisms in Chinese and the pharmacokinetics of tolbutamide. AB - AIM: To study the relationship between P450 2C9 genetic polymorphisms and the pharmacokinetics of tolbutamide in Chinese subjects. METHODS: P450 2C9 genotype was determined by oligonucleotide microarray. Using tolbutamide as a probe of P450 2C9 activity, P450 2C9 phenotype in 20 healthy individuals expressing the P450 2C9*1/*1, *1/*3 and *3/*3 genotypes were evaluated. After administration of 500 mg tolbutamide, plasma and urine samples were collected from each subject over a 24-h period. The tolbutamide and its metabolites' concentrations in human plasma and urine were determined by solid-phase extraction and HPLC. RESULTS: Tolbutamide AUC0-->infinity increased by 20 and 116%, and T(1/2) increased by 60 and 813%, respectively, in subjects expressing the P4502C9*1/*3 and *3/*3 genotypes compared with *1/*1 subjects. Reductions in tolbutamide oral clearance (68 and 11%) and formation clearance (39 and 3%) were detected in the *1/*3 and *3/*3 individuals, respectively, compared with */*1 subjects. CONCLUSION: The P450 2C9 activity was significantly reduced in *1 heterozygotes compared with *1 homozygotes, and the metabolism of tolbutamide was more severely impaired in *3/*3 individuals compared with those expressing *1/*3. Using tolbutamide as a P450 2C9 probe, P450 2C9 genotype was the major determinant of P450 2C9 phenotype. PMID- 15896242 TI - Cost considerations of the new fixed combinations for glaucoma medical therapy. AB - OBJECTIVE: To compare the costs of the new fixed combinations for glaucoma medical therapy. METHODS: The studied drugs were: Cosopt (5-mL bottle), Combigan (5-mL bottle) and Xalacom (2.5-mL bottle). Five bottles of each drug were obtained from pharmacies, and the medications lot numbers were recorded. To calculate the drop volume, 10 drops and 1 mL of each bottle were weighed with a digital precision scale. Drop volume was calculated by the relation between volume and weight. The cost of each bottle of medication was determined from the average retail price in Canada. The prices were obtained in Canadian dollars (dollars). RESULTS: The drops of Cosopt (39.60 +/- 0.45 microL) were considerably larger than the drops of Combigan (33.75 +/- 0.60 microL) and Xalacom (30.87 +/- 0.37 microL). The average number of drops per millilitre varied from 25.25 +/- 0.29 (Cosopt) to 32.40 +/- 0.39 microL (Xalacom). Combigan presented the lowest daily cost (dollars 0.87 +/- 0.02) followed by Xalacom (dollars 1.09 +/- 0.01) and Cosopt (dollars 1.22 +/- 0.01). The average cost by year varied from dollars 316.75 +/- 5.59 (Combigan) to dollars 445.96 +/- 5.16 (Cosopt), with a total difference of dollars 129.21 per year of treatment. CONCLUSIONS: There was a statistically significant difference in average drop size and cost among the three studied drugs. Combigan presented the lowest daily cost followed by Xalacom and Cosopt. PMID- 15896243 TI - Stability of doxorubicin combined with Radioselectan, a contrast agent, for chemoembolization. AB - OBJECTIVE: To assess the stability of doxorubicin combined with Radioselectan. METHODS: Solutions of doxorubicin 5 mg/mL were prepared from commercially available 50 mg powder with 10 mL of Radioselectan. They were stored in glass syringes at 4, 25 and 45 degrees C. The concentrations of doxorubicin were determined using a stability-indicating high-performance liquid chromatography method. The initial and final pHs of solutions were compared. The times (t90) needed for doxorubicin to fall to 90% of its initial concentration were calculated by a linear regression analysis. RESULTS: The t90 [95% confidence limits] were 79 [75-83], 56 [53-59] and 22 [21-23] hours for the solutions stored at 4, 25 and 40 degrees C respectively. The initial pH of the solutions stored at 4, 25 and 40 degrees C were 6.52, 6.50 and 6.51 respectively. The final pH of solutions stored at 25 and 40 degrees C decreased significantly by 0.3 and 0.9 respectively. No change of pH solution stored at 4 degrees C was observed. CONCLUSION: Doxorubicin combined with Radioselectan stored at room temperature is stable for 48 h. PMID- 15896244 TI - Use of six main drug therapeutic groups across educational groups: self-reported survey and prescription records. AB - OBJECTIVES: To assess whether the use of six main therapeutic groups was congruent with the occurrence of related diseases across educational groups. METHODS: Two data sources were analysed: (i) Interview data from The Danish Health and Morbidity Survey 2000 on a representative sample of the Danish population ages 16 years and above (n = 16,690); (ii) Prescription records linked to a health survey on a representative sample of the population of Funen County 2000-2001 (n = 3,422). The use of six therapeutic main groups (ATC groups A, B, C, M, N and R) and related diseases in educational groups was analysed by indirect standardization. Age and gender standardized prevalence ratios (SPRs) and 95% confidence intervals were calculated on the basis of the total study population. RESULTS: In general, respondents in the two least educated groups used medicines more frequently and a higher proportion of them reported the related disease than could be expected from indirect standardization. The opposite picture appeared for respondents in the two highest educational groups (SPR < 100). The overall patterns were similar for the six medicine groups, although some of the SPRs were not significant. CONCLUSION: The results show the uneven distribution of disease in the general population. The distribution of medicine use generally followed this pattern, which means that those in the greatest medical need used the most medicine. Hence, individual co-payment for medicine did not seem to be a barrier to access to medicine in any of the educational groups. PMID- 15896245 TI - TGF-beta1 mRNA expression in liver biopsy specimens and TGF-beta1 serum levels in patients with chronic hepatitis C before and after antiviral therapy. AB - BACKGROUND AND OBJECTIVE: Transforming growth factor (TGF)-beta1 is the best characterized profibrogenic cytokine. TGF-beta1 increases the production of extracellular matrix proteins and their receptors and inhibits the synthesis of matrix degrading proteolytic enzymes. We undertook this study to simultaneously evaluate the effect of interferon alpha 2b plus ribavirin therapy on TGF-beta1 daily serum levels and on mRNA TGF-beta1 expression in liver biopsy specimens from 60 patients with chronic hepatitis C. METHODS: Serum levels of TGF-beta1 were measured by ELISA. The levels of the RNAs in liver biopsy specimens were measured by quantitative reverse transcriptase polymerase chain reaction. After treatment, patients were divided into two groups: 34 responders [undetectable hepatitis C virus (HCV)-RNA, normal ALT levels, decrease in histology activity index compared with pretreatment liver biopsy] and 26 non-responders (detectable HCV-RNA, elevated ALT levels, no decrease in the histology activity index). RESULTS AND DISCUSSION: In patients with hepatitis C, the 'responders' to the antiviral treatment showed significant decreases in both mean daily serum TGF beta1 levels and mRNA TGF-beta1 expression in the liver biopsy specimens. The 'non-responders' serum TGF-beta1 concentrations did not change significantly, but the mRNA TGF-beta1 expression did. CONCLUSION: Both serum TGF-beta1 concentration and mRNA TGF-beta1 expression in liver biopsy specimens may be useful as prognostic markers in patients with hepatitis C undergoing antiviral therapy. PMID- 15896246 TI - Assessing the accuracy of a computerized decision support system for digoxin dosing in primary care: an observational study. AB - BACKGROUND: This study was carried out as part of a European Union funded project (PharmDIS-e+), to develop and evaluate software aimed at assisting physicians with drug dosing. A drug that causes particular problems with drug dosing in primary care is digoxin because of its narrow therapeutic range and low therapeutic index. OBJECTIVES: To determine (i) accuracy of the PharmDIS-e+ software for predicting serum digoxin levels in patients who are taking this drug regularly; (ii) whether there are statistically significant differences between predicted digoxin levels and those measured by a laboratory and (iii) whether there are differences between doses prescribed by general practitioners and those suggested by the program. METHODS: We needed 45 patients to have 95% Power to reject the null hypothesis that the mean serum digoxin concentration was within 10% of the mean predicted digoxin concentration. Patients were recruited from two general practices and had been taking digoxin for at least 4 months. Exclusion criteria were dementia, low adherence to digoxin and use of other medications known to interact to a clinically important extent with digoxin. RESULTS: Forty five patients were recruited. There was a correlation of 0.65 between measured and predicted digoxin concentrations (P < 0.001). The mean difference was 0.12 microg/L (SD 0.26; 95% CI 0.04, 0.19, P = 0.005). Forty-seven per cent of the patients were prescribed the same dose as recommended by the software, 44% were prescribed a higher dose and 9% a lower dose than recommended. CONCLUSION: PharmDIS-e+ software was able to predict serum digoxin levels with acceptable accuracy in most patients. PMID- 15896247 TI - Ketoconazole increases plasma concentrations of antimalarial mefloquine in healthy human volunteers. AB - BACKGROUND: Antimalarial mefloquine has a structure related to quinine. The major metabolite of quinine is 3-hydroxyquinine formed by cytochrome P450 3A4 (CYP3A4). Ketoconazole, a potent inhibitor of CYP3A4, is known to markedly increase plasma concentrations of various co-administered drugs including quinine. OBJECTIVE: To assess the effect of ketoconazole on plasma concentrations of mefloquine in healthy Thai male volunteers. METHODS: In an open, randomized two-phase crossover study separated by a 1-month period, eight healthy Thai male volunteers received a single oral dose of 500 mg mefloquine alone or co-administration with 400 mg/day ketoconazole orally for 10 days. Serial blood samples were collected at specific time points for a 56-day period. Plasma mefloquine and mefloquine carboxylic metabolite concentrations during 56 days were measured by a modified and validated high-performance liquid chromatographic method with UV detection. RESULTS: Co-administration with ketoconazole markedly increased the mean values of mefloquine AUC0-t, t(1/2), and Cmax when compared with mefloquine alone by 79% (P < 0.001), 39% (P < 0.05) and 64% (P < 0.001) respectively. The AUC0-t , and Cmax of mefloquine carboxylic acid metabolite were decreased by 28% (P < 0.05) and 31% (P < 0.05), respectively when compared with mefloquine alone. CONCLUSIONS: Co-administration with ketoconazole increased plasma mefloquine concentrations in healthy human volunteers. One of possible mechanisms of the increase in plasma mefloquine concentrations may be the result of the inhibition of CYP3A4 by ketoconazole. In case of mefloquine is co-administered with ketoconazole, drug-drug interactions should be recognized and the dose of mefloquine should be adjusted to maximize the therapeutic efficacy and to reduce the cost of therapy. PMID- 15896248 TI - Proposed mechanisms and preventative options of Jarisch-Herxheimer reactions. AB - OBJECTIVE: To review the aetiologies and preventative methods associated with Jarisch-Herxheimer reactions (JHR). DATA SOURCES: Ovid Medline (1966-June Week 1 2004) was utilized to assess biomedical literature; a review of the bibliographies of articles was also performed. DATA SYNTHESIS: JHR often occurs with the treatment of spirochete infections. However, the mechanism by which the reaction takes place is not clearly defined. CONCLUSION: Studies suggest with conflicting evidence that the JHR is caused by release of endotoxin-like material from the spirochete as well as cytokine elevation in the body. It appears the type of drug and the rate of spirochete clearance from the body have little effect on the incidence of the reaction. Many pretreatment options have been explored with limited efficacy with the exception of anti-tumour necrosis factor antibodies. PMID- 15896249 TI - A study of the management of erectile dysfunction in general practice. AB - BACKGROUND AND OBJECTIVES: The Department of Health issued guidelines for the NHS treatment of erectile dysfunction (ED) with phosphodiesterase type 5 inhibitors (PDE 5 inhibitors) in 1999. There has been an increasing trend in the prescribing of PDE 5 inhibitors within Bebington and West Wirral Primary Care Trust (PCT) over the 3-year period from February 2001 to January 2004. The objective of the study was to investigate implementation of Government guidelines on prescribing of PDE 5 inhibitors for ED and the cost of prescribing outside these guidelines. METHODS: Practice data were collected for all patients prescribed a PDE 5 inhibitor in 16 surgeries within Bebington and West Wirral Primary Care Trust, from November 2002 to December 2003. The data were evaluated with respect to adherence to UK Government guidelines. Analysis was made on the cost to the PCT with respect to treatment provided outside the guidelines. RESULTS AND DISCUSSION: Prescribing for 78% of patients was within Government guidelines. With respect to frequency of prescribing, 89% of patients in the PCT received less than or equal to the recommended frequency of one tablet per week. The percentage range for practices was 67-100%. The cost to the PCT for PDE 5 inhibitor treatment provided outside the guidelines was 19,060 pounds sterling over the period of study. CONCLUSION: Prescribers generally follow Government guidelines, however, stricter adherence to guidelines could result in more efficient use of National Health Service resources. PMID- 15896252 TI - A call for deeper scrutiny of mental health care for people in Australian immigration detention centres. PMID- 15896253 TI - The experience and views of mental health nurses regarding nursing care delivery in an integrated, inpatient setting. AB - Positive and effective consumer outcomes hinge on having in place optimal models of nursing care delivery. The aim of this study was to ascertain the experience and views of mental health nurses, working in hospitals in an area mental health service, regarding nursing care delivery in those settings. Surveys (n = 250) were sent to all mental health nurses working in inpatient settings and 118 (47%) were returned. Results showed that the quality of nursing care achieved high ratings (by 87%), and that two-thirds of respondents were proud to be a mental health nurse and would choose to be a mental health nurse again. Similarly, the majority (71%) would recommend mental health nursing to others. Concern was, however, expressed about the continuity and consistency of nursing work and information technology resources. Nurses with community experiences rated the importance of the following items, or their confidence, higher than those without previous community placements: the importance of interdisciplinary teamwork; the importance of participating in case review; the importance of collaborating with community staff; confidence in performing mental state examinations; and confidence in collaborating with community staff, suggesting that this placement had positive effects on acute care nursing. PMID- 15896254 TI - The attitudes, knowledge and skills needed in mental health nurses: the perspective of users of mental health services. AB - Knowledge about the practice and roles of mental health nurses in New Zealand is currently limited. In a sociopolitical climate where the views of users of mental health services are increasingly incorporated into education and the planning and delivery of services, there is a need for research that reflects the perspective of users. In this study a qualitative descriptive methodology with focus group interviews was used to explore with users of mental health services, the attitudes, knowledge and skills that they need in mental health nurses. Users of mental health services valued the therapeutic work of mental health nurses, and identified positive attitudes towards users of mental health services as essential in mental health nurses. However, they did not consistently experience a therapeutic approach in their interactions with mental health nurses. PMID- 15896255 TI - Factors that influence Asian communities' access to mental health care. AB - This paper presents the findings of a qualitative study to identify factors that influence Asian communities' access to mental health care and how mental health care is delivered to them. Semistructured interviews were completed with Asian community members/leaders and health-care professionals. Content analysis identified major themes. Participants also completed a demographic data sheet. The research aimed to provide health professionals with an increased understanding of the values and beliefs held by people from Asian communities regarding the cause and treatment of mental illness. Data analysis identified six main themes that influenced Asian communities' access to mental health care and how mental health care is delivered to them. They were: shame and stigma; causes of mental illness; family reputation; hiding up; seeking help; and lack of collaboration. The findings highlighted that people from Asian communities are unwilling to access help from mainstream services because of their beliefs, and that stigma and shame are key factors that influence this reluctance. The findings also highlight that the mental health needs of refugee women are significant, and that they comprise a vulnerable group within Australian society. PMID- 15896256 TI - Cognitive behaviour therapy within acute mental health care: a critical appraisal. AB - This paper examines the evidence for the use of cognitive behavioural therapy (CBT) within acute mental health-care settings. Through the provision of a literature review the authors critique the research into the effectiveness of CBT when delivered within acute mental health-care settings, to clients who are acutely psychotic. The review concludes with recommendations for future research to expand the current evidence base. PMID- 15896257 TI - Nursing student attitudes to psychiatric nursing and psychiatric disorders in New Zealand. AB - Attitudes towards mental illness and psychiatric nursing can have far-reaching consequences for patients and the perceived desirability of this as a future career choice. Attitudes can be both brought into training and affected by training experiences, although the direction of these associations is unclear. Using a cross-section design, this study investigates the associations between attitudes, demographic variables, mental illness exposure, and career aspirations in 164 nursing students. Year of training and area of intended practice independently predicted attitudes. These findings have important implications for nursing training. They demonstrate the need for specifically focusing training to target the students with the most negative attitudes. PMID- 15896258 TI - Promoting a sustainable mental health nursing workforce: an evaluation of a transition mental health nursing programme. AB - The recruitment and retention of the mental health nursing workforce has been identified as a major issue both nationally and internationally. Transition to practice programmes has been identified as an important strategy in addressing these issues. There is, however, a paucity of literature addressing the potential or effectiveness of transition programmes in achieving these aims. This paper reports the findings of a survey administered to registered nurses at the commencement and completion of the Transition Programme into Mental Health Nursing, in Sydney, Australia. The findings suggest a high level of satisfaction with the programme. The results are directing the ongoing development of clinical placements, clinical supports, education programmes and recruitment and retention initiatives for nurses within the Central Sydney Area Mental Health Service. PMID- 15896259 TI - The validity and reliability of the World Health Organization Mental Disorders Checklist for use in a telehealth clinic in Hong Kong. AB - This research aimed to test the validity and reliability of the 'World Health Organization Mental Disorders Checklist' for use in a telehealth clinic in Hong Kong. The Checklist adopted four subscales: (i) depression; (ii) anxiety; (iii) alcohol use disorders; and (vi) functioning and disablement, and was translated from English into Chinese. It was validated by a panel of five experts to confirm its content validity (content validity index = 0.98) and cultural appropriateness in Hong Kong. The reliability of the checklist was supported by the findings of a test-retest procedure (Pearson correlation = 0.66-0.88, P < 0.01), internal consistency reliability (Cronbach's alpha = 0.54-0.83), and interrater reliability (Kendall's coefficient of concordance = 0.58-1.00, P < 0.01) involving a sample of 197 subjects from one telehealth clinic in Hong Kong. PMID- 15896260 TI - Interpersonal psychotherapy: an effective psychotherapeutic intervention for mental health nursing practice. AB - Interpersonal psychotherapy (IPT) is well-recognized for the treatment of depression with many efficacy trials supporting its use. However, there is little discussion of its use in mental health nursing practice. This paper explores how IPT can be a useful intervention for mental health nursing practice and demonstrates the process of IPT with reference to a case study. The case study illustrates how IPT facilitates a shift for one woman from a passive subject position to a more self-assertive one. This shift was facilitated by identifying how she was constructing herself in relation to others by utilizing tactics of passivity and avoidance of conflict. The development of more satisfying subject positions facilitated an improvement in mood and recovery from depression. PMID- 15896261 TI - The role of the registered nurse in an acute mental health inpatient setting in New Zealand: perceptions versus reality. AB - Following the closure of the large psychiatric institutions in New Zealand, there is an increasing demand for limited beds in acute inpatient facilities for acutely mentally ill patients. This change in location and downsizing of acute inpatient beds has challenged traditional roles of mental health nursing, resulting in confusion over what roles mental health nurses should now perform in the new context of care. This study compared the perceptions that registered psychiatric nurses have of their roles with their actual practice. This qualitative descriptive exploratory study observed nursing practice on three selected wards and used focus group interviews to establish from registered nurses what they perceived their roles to be. A key finding of this study was that many of the nursing roles related to delivering care from a crisis management perspective, which covers aspects such as assessment, stabilization of symptoms and discharge planning. Participants also believed that the therapeutic relationship was a fundamental role in inpatient care. Nurses used any opportunity to make it a reality such as kitchen organization, medications, or dealing with a challenging patient. This study highlighted the complexity of the roles that nurses performed and went some way to give voice to what at times seems an invisible practice. PMID- 15896262 TI - The phenomenon of resilience in crisis care mental health clinicians. AB - The purpose of this study, undertaken in 2003, was to explore the phenomenon of resilience as experienced by Australian crisis care mental health clinicians working in a highly demanding, complex, specialized and stressful environment. For the purpose of this research, the term 'resilience' was defined as the ability of an individual to bounce back from adversity and persevere through difficult times. The six participants for this study were drawn from Melbourne metropolitan mental health organizations - the disciplines of nursing, allied health and medicine. A number of themes were explicated from the participants' interview transcripts - Participants identified the experience of resilience through five exhaustive descriptions, which included: 'The team is a protective veneer to the stress of the work'; Sense of self; Faith and hope; Having insight; and Looking after yourself. These exhaustive descriptions were integrated into a fundamental structure of resilience for clinicians in this role. The study's findings have the potential to inform organizations in mental health to promote resilience in clinicians, with the potential to reduce the risk of burnout and hence staff attrition, and promote staff retention and occupational mental health. PMID- 15896263 TI - Sexual health knowledge and risk behaviour in young people with first episode psychosis. AB - The sexual health of clients has been poorly addressed by mental health service providers and sexually transmissible infection (STI) incidence rates are increasing among young people. A self-report questionnaire was used to gather information in order to describe and to conduct a cross-sectional analysis of sexual health knowledge, risk behaviour and STI screening history among young people who have experienced a first episode of psychosis, who present to community-based early psychosis programmes in south-eastern Sydney. STI knowledge was fair and sexual risk practice knowledge was poor regarding anal and oral sex. Women reported significantly more sexual partners than men. When those young people who had had unsafe sex attended for STI screens only, 5% received a complete screen. The survey result indicates a need for sexual health screening education to be conducted for clients. Against a background of escalating rates of HIV, STIs and high risk-taking behaviours among young people, it is essential that mental health staff are provided with the skills and education to address sexual health and harm minimization issues. PMID- 15896264 TI - Brain regional alpha-[11C]methyl-L-tryptophan trapping correlates with post mortem tissue serotonin content and [11C]5-hydroxytryptophan accumulation. PMID- 15896265 TI - Molecular genetics support Gray's personality theory: the interaction of COMT and DRD2 polymorphisms predicts the behavioural approach system. AB - The present study provides the first direct molecular genetics support for Gray's Reinforcement Sensitivity Theory (RST), which is one of the most influential biologically oriented personality theories. It was investigated whether the DRD2 TaqIA and the COMT polymorphisms were related to the dimensions of Gray's personality theory, as measured by the Carver and White BIS/BAS scales. In a sample of 295 healthy subjects results revealed significant DRD2xCOMT interactions (i.e. epistasis) for the total BAS scale (related to positive emotionality) and for the subscales Drive (D) and Fun Seeking (FS). High BAS scores were observed if the catabolic enzyme activity and the D2 receptor density as indicated by the two polymorphisms were in disequilibrium, i.e. in the presence of the Val-/A1- (low enzyme activity/high receptor density) or the Val+/A1+ (high enzyme activity/low receptor density) alleles. In a random subsample (n=48), it could be demonstrated that those allele combinations of COMT and DRD2 associated with high BAS scores also had significantly lower prolactin levels under resting conditions, indicating high dopamine activity, compared to those allele combinations with low BAS scores. Furthermore, two-way interactions of DRD2 TaqIAxsmoking status and of the Met allele of COMTxsmoking status on FS and Metxgender on BIS could be shown. PMID- 15896266 TI - Focus on clozapine: a new explanation for its atypical character. PMID- 15896267 TI - Labeling of human insulin-like growth factor-I eukaryotic expression vector with green fluorescent protein. AB - OBJECTIVE: To label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects. METHODS: GFP cDNA was inserted into pcDNA(3.1)-hIGF 1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGI was transfected into NIH 3T3 cells with the help of lipofectamine. RESULTS: Enzyme digestion and agarose gel electrophoresis analysis revealed that pcGI vector contained correct GFP and hIGF-I cDNA. Expression of hIGF-1 and GFP was confirmed in transfected NIH 3T3 cells by immunocytochemical analysis and fluorescence microscopy. CONCLUSIONS: hIGF-I eukaryotic expression vector has been successfully labeled with GFP. PMID- 15896268 TI - Study of the effects of mild hypothermia on cerebral PO2, PCO2 and pH and body temperature in patients with acute severe head injury. AB - OBJECTIVE: To study the effects of mild hypothermia on cerebral oxygen partial pressure, carbon dioxide partial pressure, pH and body temperature (PbrO2, PbrCO2, pHbr and BT) in patients with acute severe head injury. METHODS: Thirty eight patients with acute severe head injury were treated with mild hypothermia, meantime PbrO2, PbrCO2, pHbr and BT were monitored in order to study the changes of PbrO2, PbrCO2, pHbr and BT. RESULTS: In patients with acute head injury, mild hypothermia obviously increased PbrO2, decreased PbrCO2 and CO2 accumulation and acidosis in brain tissue. BT was 1-1.5 deg. higher than rectal temperature (RT) after injury. The BT and RT were decreased when the patients were treated with mild hypothermia, but at the same time the difference between BT and RT was increased. CONCLUSIONS: In patients with acute severe head injury the direct monitoring of PbrO2, PbrCO2, pHbr and BT was safe and reliable, and is helpful in estimating prognosis and mild hypothermia therapy. PMID- 15896269 TI - Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene. AB - OBJECTIVE: To construct an eukaryotic expression vector carrying rat brain derived neurotrophic factor receptor trkB gene. METHODS: Using the total RNA isolated from rat brain as template, the trkB gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers which contained the restrictive sites of EcoR I and BamH I. The amplified fragment of trkB gene was digested with EcoR I and BamH I, and then subcloned into cloning vector pMD18-T and expression vector pEGFP-C2 respectively. The recombinant plasmids were identified by restriction endonuclease enzyme analysis and PCR. RESULTS: The amplified DNA fragment was about 1461 bp in length. Enzyme digestion and PCR analysis showed that the gene of trkB had been successfully cloned into vector pMD18-T and pEGFP-C2. CONCLUSIONS: The trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2. PMID- 15896270 TI - Comparative study on effects of burn-blast combined injury and burn-firearm combined injury complicated with seawater immersion on vascular endothelial cells. AB - OBJECTIVE: To comparatively study the effects and mechanisms of burn-blast combined injury and burn-firearm combined injury complicated with seawater immersion on vascular endothelial cells. METHODS: A total of 40 healthy adult hybrid dogs of both sexes, weighing 12-15 kg, were used in this study. Randomly selected 20 dogs were established as models of burn-blast combined injury (the burn-blast injury group) and the other 20 dogs as models of burn-firearm combined injury (the burn-firearm injury group). Then the wounds of all the dogs were immediately immersed in seawater for 4 hours, and then they were taken out from the seawater. Blood samples were withdrawn from the central vein of the dogs before injury, and at 4, 7, 10, 20, and 28 hours after injury to measure the circulating endothelial cells and the von Willebrand factor. RESULTS: Circulating endothelial cells increased significantly at 4 hours after injury in all the dogs. But they reached peak at 7 hours after injury in the burn-blast injury group and at 28 hours after injury in the burn-firearm injury group. The changes of circulating endothelial cells in the burn-blast injury group were significantly different from those in the burn-firearm injury group at 4, 7, 20, and 28 hours after injury (P < 0.01). The von Willebrand factor reached peak at 4 hours after injury in the burn-blast injury group and at 28 hours in the burn firearm injury group. The changes of von Willebrand factor in the burn-blast injury group were significantly different from those in the burn-firearm injury group at 4, 20, and 28 hours after injury (P < 0.01). CONCLUSIONS: In burn-blast injury combined with seawater immersion, the vascular endothelial cells changed most significantly at 4 hours or 7 hours after injury, while burn-firearm injury combined with seawater immersion have the same at 20 hours or 28 hours after injury. PMID- 15896271 TI - Changes of osteocalcin and IGF-I during bone lengthening. AB - OBJECTIVE: To examine the changes of osteocalcin and insulin-like growth factor I (IGF-I) during bone lengthening, and to clarify the mechanism of bone healing. METHODS: Thirty-two shepherd dogs were divided into five groups randomly. Their tibiae were lengthened by Ilvzarov's external fixator at the rate of 1 mm/day. The lengthening area was the experimental side and the opposite side was the control. Samples were obtained on the 2nd, 4th, 6th, 8th, 12th weekend respectively. The samples were defatted, dried, powdered, centrifuged and measured by radioimmunoassay. RESULTS: The osteocalcin concentration increased at the subsequent periods, but it was significantly lower in the experimental side than that of the control side, P<0.05 and the IGF-I concentration was not significantly lowered. CONCLUSIONS: Different noncollagenous bone growth factors may be different at different stage. PMID- 15896272 TI - External fixation and bone grafting for collapsed and comminuted distal radius fracture. AB - OBJECTIVE: To study the therapeutic effect of collapsed and comminuted distal radius fracture. METHODS: Twenty-six patients with collapsed and comminuted distal radius fracture were hospitalized from July 1998 to June 2003. All fractures were treated by the methods of open reduction, sustained bone grafting and passing joint external fixator to restore the anatomic shape of distal radius. RESULTS: All 26 cases were followed up, and the results showed that the fractures have been united radiographically. The joint surfaces were intact and there was no length discrepancy occurred in patient's radius. The average volar tilt was 6 to 15 degrees and the average ulnar tilt was 18 to 25 degrees. According to the Dieust criterion, 19 cases were rated as excellent and 7 as good. CONCLUSIONS: The method that applying passing joint external fixator and bone grafting for the treatment of collapsed and comminuted distal radius fracture could maintain the stability of fracture and restore the length of radius and the intact of joint surface. PMID- 15896273 TI - Lower cervical spine injury treated with lateral mass plates and pedicle screws through posterior approach. AB - OBJECTIVE: To treat injury of the lower cervical spine C6 to C7 with cervical lateral mass plates and T1 pedicle screws through posterior approach. METHODS: The data of 8 patients with lower cervical spine C6 or C7 injury (6 patients with fracture and dislocation in C6 and C7 and 2 with fracture in C7) were analyzed retrospectively in this study. For the preoperative American Spinal Injury Association (ASIA) classification, Grade C was found in 3 cases and Grade D in 5 cases. Screws were placed on the lateral masses and the first thoracic pedicle with Margerl technique. Lamina or facet bone allografting was used to achieve a long-term stability. RESULTS: All the 8 patients were followed up for 5-37 months (mean: 15 months). No operative death occurred. There were no examples of aggravation of spinal cord injury or vertebral artery injury, cerebrospinal fluid leak, nerve roots injury, screw malposition or back-out, loose of alignment or implant failure. Clinical symptoms and ASIA classification were improved in all the patients. Postoperative MRI scanning confirmed the satisfactory screw placement in all the cases. CONCLUSIONS: Lateral mass plates and pedicle screws through posterior approach are safe and beneficial for patients with lower cervical spine C6 or C7 injury. PMID- 15896274 TI - The analysis of injured and poisoned patients during the construction of urbanization and industrialization from 1998 to 2002. AB - OBJECTIVE: To analyze the causes of injury and poisoning during construction of urbanization and industrialization and to explore proper measures. METHODS: The data of the patients with injury and poisoning during construction of urbanization and industrialization treated in out hospital from 1998 to 2002 were retrospectively analyzed. RESULTS: The number of discharged patients of injury and poisoning accounted for 29.2% of the total number (49800) of discharged patients in the corresponding period in our hospital, which was greater than that of other city and county hospitals, and accounted for 94.3% of the total number (15,411) of discharged patients of the Surgery Department in the corresponding period in our hospital. Injuries caused by motor vehicle traffic accidents, cutting and piercing instruments or objects, homicide and injury purposely inflicted by other persons and accidental falls held 78.9%. The number of the inpatients in 2002 increased by 83.3% compared with that of 1998, the number of injured and poisoned inpatients increased by 76.1% and these patients aged mainly at age of 15-39 years, holding 80.1%. In a year, the peak period of trauma patients was in summer because of hot weather; the fewest in February for the floating people went home for Spring Festival holidays. CONCLUSIONS: During construction of rural urbanization and industrialization, injuries and poisoning increase evidently and are the main tasks of surgical management. So to raise its treatment level and therapeutic effect is a key point. PMID- 15896275 TI - Neurogenic pulmonary edema in head injuries: analysis of 5 cases. AB - OBJECTIVE: To review the pathophysiology and study the diagnosis and clinical management of neurogenic pulmonary edema (NPE). METHODS: The data of 5 patients who developed NPE after head injury treated in our hospital form December 1995 to May 2003 were collected and analyzed. RESULTS: The patients developed dyspnea and respiratory failure 2-8 hours after neurologic event. Four of the 5 patients presented with pink frothy sputum. Chest radiography showed bilateral diffuse infiltrations in all the 5 patients. After supportive measures such as oxygen support and pharmacologic therapy, 4 patients recovered in 72 hours and one patient died. CONCLUSIONS: The pathophysiologic mechanisms of NPE is unclear. In acute respiratory failure following head injury, NPE must be given much attention and timely and effective measures should be taken. PMID- 15896276 TI - Effects of low-intensity pulsed ultrasound in repairing injured articular cartilage. AB - OBJECTIVE: To investigate the effects of low-intensity pulsed ultrasound in repairing injured articular cartilage. METHODS: Ten adult New Zealand rabbits with bilateral full-thickness osteochondral defects on the cartilage surface of intercondylar fossas were used in this study. The wounds in the left knees were treated with low-intensity pulsed ultrasound as the experimental group. The right knees received no treatment as the control group. All the animals were killed at 8 weeks after injury and the tissues in the wounds were collected for gross appearance grading, histological grading and proteoglycan quantity. RESULTS: The scores of the gross appearance grades, histological grades and the optical density of toluidine blue of the tissues in the experimental group were significantly higher than those of the controls at 8 weeks after injury (P < 0.05). CONCLUSIONS: Low-intensity pulsed ultrasound can accelerate the repair of injured articular cartilage. PMID- 15896277 TI - Time window characteristics of cultured rat hippocampal neurons subjected to ischemia and reperfusion. AB - OBJECTIVE: To explore cell death and apoptosis in rat hippocampal neurons at different time points after ischemia, hypoxia and reperfusion injury and to elucidate time window characteristics in ischemia neuronal injury. METHODS: Hippocampal neurons were obtained from rat embryo and were cultured in vitro. The ischemia and reperfusion of cultured rat hippocampal neurons were simulated by oxygen-glucose deprivation (OGD) and recovery. OGD at different time points (0.25 h to 3.0 h) and then the same recovery (24 h) were prepared. Annexin V-PI staining and flow cytometry examined neuron death and apoptosis at different time after injury. RESULTS: After OGD and recovery, both necrosis and apoptosis were observed. At different times after OGD, there were statistically significant differences in neuron necrosis rate (P < 0.05), but not in apoptosis rate (P > 0.05). At recovery, survival rate of hippocampal neurons further decreased while apoptosis rate increased. Furthermore, apoptosis rates of different time differed greatly (P < 0.05). Apoptosis rate gradually increased with significant difference among those of different time points (P < 0.05). However, 2 h after ischemia, apoptosis rate decreased markedly. CONCLUSIONS: Apoptosis is an important pathway of delayed neuron death. The therapeutic time window should be within 2 h after cerebral ischemia and hypoxia. PMID- 15896278 TI - The clinical characteristics and therapy of syndrome of craniocerebral-cervical vertebral injury. AB - OBJECTIVE: To explore the clinical characteristics and new treatment for syndrome of craniocerebral-cervical vertebral injury. METHODS: The clinical data of 52 patients with head injury accompanied by neck injury were analyzed retrospectively. RESULTS: Craniocerebral injury could result in damage to cervical vertebrae, muscles, vessels and nerves, and even cause vertebral artery injury, which may lead to insufficient blood-supply of vertebral-basal artery. All patients were treated with cervical vertebral traction and the results were good. CONCLUSIONS: Acute craniocerebral injury with symptom of insufficient blood supply of vertebral-basal artery, evident neurosis and atlas-axis half dislocation in X-ray should be treated by cervical vertebral traction, which will yield better outcome. PMID- 15896279 TI - Pathological study of rabbits' femoral arteries subjected to gunshot wounds combining with seawater immersion. AB - OBJECTIVE: To investigate pathological characteristics of gunshot wounds concomitant seawater immersion in rabbits' femoral arteries. METHODS: Thirty rabbits were divided randomly into 3 groups: simple gunshot-wound group (Group I, n = 10), gunshot wound with seawater immersion for 30 mins (Group II, n = 10), and 60 mins group (Group III, n = 10). Femoral arteries were impacted by 0.38 g steel spheres fired with a 7.62 mm rifle. After being wounded, rabbits in Groups II and III were immersed in seawater for 30 or 60 mins, but those in Group I were not. At 2, 4, 6, 8, 12 hours following injury, a 40 mm segment of the artery on each side of the gunshot point were excised and observed by light and electron microscopy. RESULTS: The patterns of arterial injuries were mainly contusion and transection. Completely transected artery was classified as primary-wound-tract area, contused area and shocked area. Compared with those in Group I, the primary wound-tract and contused areas in Group II manifested obvious swelling in the arterial wall especially at the outside 2/3 of the media. Vacuolar structures were often seen in smooth muscle cells of the media. Intercellular space among the smooth muscle cells were filled with homogeneous acidophilic substances. Deep rugae among endothelial cells flattened or rugal folds lost their longitudinal orientation, and marked fibrin and platelet deposition were noticed. No significant difference was detected between Group II and III. The pathological changes in the shocked area were similar in 3 groups. CONCLUSIONS: For gunshot wounds concomitant seawater immersion in rabbits' femoral arteries, there was a marked swelling of cells and intercellular space in primary-wound-tract area and contusion area. The influence of these pathological changes on surgical reparation deservers further study. PMID- 15896280 TI - Surgical treatment for traumatic transection of left common carotid artery. AB - Injury to the supraaortic artery is a rare event, with poor prognosis and high mortality. Improvement of the outcome may lie on the combination of several aspects, including better pre-hospital care, use of emergency cardiopulmonary bypass (CPB), improved surgical techniques and facilities, and advanced postoperative intensive care. Some researchers emphasized the importance of emergency CPB in the treatment and thought that it was responsible mainly for the improved outcome. However, there exists controversies about it. In this article, we reported that a patient with life-threatening hemorrhage due to traumatic transection of the left common carotid artery, who was admitted to our hospital in July 2003, was treated successfully with operations with help of emergency CPB and systemic hypothermia. PMID- 15896281 TI - Cognitive-behavioral therapy for ADHD in medication-treated adults with continued symptoms. AB - The purpose of the present study was to examine the potential efficacy, patient acceptability, and feasibility of a novel, cognitive-behavioral therapy (CBT) for adults with attention-deficit hyperactivity disorder (ADHD) who have been stabilized on medications but still show clinically significant symptoms. Thirty one adults with ADHD and stable psychopharmacology for ADHD were randomized to CBT plus continued psychopharmacology or continued psychopharmacology alone. Assessments included ADHD severity and associated anxiety and depression rated by an independent evaluator (IE) and by self-report. At the outcome assessment, those who were randomized to CBT had lower IE-rated ADHD symptoms (p < .01) and global severity (p < .002), as well as self-reported ADHD symptoms (p < .0001) than those randomized to continued psychopharmacology alone. Those in the CBT group also had lower IE-rated and self-report anxiety (p's < .04), lower IE-rated depression (p < .01), and a trend to have lower self-reported depression (p = .06). CBT continued to show superiority over continued psychopharmacology alone when statistically controlling levels of depression in analyses of core ADHD symptoms. There were significantly more treatment responders among patients who received CBT (56%) compared to those who did not (13%) (p < .02). These data support the hypothesis that CBT for adults with ADHD with residual symptoms is a feasible, acceptable, and potentially efficacious next-step treatment approach, worthy of further testing. PMID- 15896282 TI - Misperception of sleep can adversely affect daytime functioning in insomnia. AB - This experiment was designed to investigate the relationship between subjective perception of sleep and daytime processes in primary insomnia. Twenty-two individuals with primary insomnia received positive or negative feedback about their sleep, immediately on waking, on three consecutive mornings. The positive feedback was that last night's sleep was good quality. The negative feedback was that last night's sleep was poor quality. Objective sleep on each of the three nights was estimated by actigraphy and did not differ across the three nights or the two feedback conditions. Negative feedback (based on 32 nights of data) was associated with more negative thoughts, sleepiness, monitoring for sleep-related threat, and safety behaviours during the day, relative to positive feedback (based on 34 nights of data). These results indicate that the impaired daytime functioning reported by insomnia patients is maintained, at least in part, by subjective perception of sleep. PMID- 15896283 TI - Religiosity and obsessive-compulsive behavior in Israeli Jews. AB - Individual differences in obsessive-compulsive (OC) behavior in various cultures correlate with religiosity. The current paper explored the so far unstudied relationship between religiosity and OC behavior in Israeli Jews. Two studies were conducted. Study 1 focused on the relationship between religiosity and OC behavior in a representative sample of Israeli students. Study 2 focused on religious change and OC behavior in a non-random sample of 31 individuals who had become more religious (the MR group), and 30 individuals who were less religious (the LR group) than their parents. Instruments used were the Maudsley obsessive compulsive inventory (MOCI), the student religiosity questionnaire, and questions about parental home observance, upbringing, and changes in religiosity. In the first study, no association was found between religiosity and OC behavior. Religiosity was related to some degree to perfectionism and to the parental attitude to upbringing. In the second study, a significant difference was observed between the MR and the LR groups on OC behavior as measured by the MOCI. Conclusion, among Israeli Jews a lot of religious observance is non-reflective, and is not associated with individual differences in personality or OC symptoms. Those who undergo religious change may do so in response to their behavioral propensities. One such path is that the more OC become MR, and the less OC less religiously observant. PMID- 15896284 TI - Trichotillomania: impact on psychosocial functioning and quality of life. AB - This study explored the impact of hair pulling on psychosocial functioning for patients diagnosed with trichotillomania (TTM; n = 28). TTM patients were compared to age and gender-matched groups of psychiatric patients without TTM (n = 28) and nonpsychiatric control volunteers (NC, n = 28) on measures of psychological distress, functioning/quality of life, and self-esteem. Results indicated that TTM patients reported more severe psychosocial impairments than did NC volunteers; however, these differences were mediated by differences in level of depression. Regression analyses indicated significant relationships between some measures of psychosocial functioning and severity of hair pulling, independent of level of depression. Finally, an interview of the impact of hair pulling on 6 domains of daily functioning (negative affect/negative self evaluations, grooming, recreational activities, social interaction, work/housework productivity, and physical health) indicated common and wide ranging impairments for both lifetime and current (i.e., past week) ratings. These results highlight the importance of promoting and improving resources for the clinical care of TTM patients, and provide some directions for clinicians to enhance assessment of interference caused by TTM. PMID- 15896285 TI - Perception of control over anxiety mediates the relation between catastrophic thinking and social anxiety in social phobia. AB - Cognitive models of social phobia (social anxiety disorder) assume that individuals with social phobia experience anxiety in social situations in part because they overestimate the social cost associated with a potentially negative outcome of a social interaction. Some emotion theorists, on the other hand, point to the perception of control over anxiety-related symptoms as a determinant of social anxiety. In order to examine the relationship between perceived emotional control (PEC), estimated social cost (ESC), and subjective anxiety, we compared three alternative structural equation models: Model 1 assumes that PEC and ESC independently predict social anxiety; Model 2 assumes that ESC partially mediates the relationship between PEC and anxiety, and Model 3 assumes that PEC partially mediates the relationship between ESC and anxiety. We recruited 144 participants with social phobia and administered self-report measures of estimated social cost, perceived anxiety control, and social anxiety. The results support Model 3 and suggest that "costly" social situations are anxiety provoking in part because social phobic individuals perceive their anxiety symptoms as being out of control. PMID- 15896286 TI - Hoarding and its relation to obsessive-compulsive disorder. AB - Although hoarding is observed in some patients with obsessive-compulsive disorder (OCD), it has not been shown to share a specific relation with OCD. Across two studies, we found that (a) whereas the classic OCD symptoms of checking, rituals, and contamination intercorrelated consistently strongly with one another, hoarding related only moderately to both these OCD symptoms and to depression; (b) OCD patients were distinguished from both other patients and non-patients by classic OCD symptoms, but not by hoarding; and (c) whereas OCD symptoms showed consistent relations with Negative Affect, hoarding largely was uncorrelated with this dimension. These results do not support a specific OCD-hoarding relation but rather call into question the trend of considering it a specific symptom of OCD. PMID- 15896287 TI - Cognitive inhibition in trichotillomania and obsessive-compulsive disorder. AB - Cognitive inhibition was investigated in 21 trichotillomania (TTM), 21 obsessive compulsive disorder (OCD) and 26 healthy control (HC) participants using a block cued directed forgetting task. After encoding a word list, participants were instructed to intentionally FORGET these words and to REMEMBER another word list. Both lists included TTM-related and neutral (kitchen-related) words in equal proportions, with the TTM-related words generally of negative valence and the kitchen-related words generally of neutral valence for all participants. A superior free recall of REMEMBER versus FORGET words suggests intact cognitive inhibition. The performance of OCD participants indicated a specific deficit in inhibiting the retrieval of information with negative valence, which was not found in TTM. HC participants, in contrast to TTM and OCD participants, were rather inattentive to negative information. In conclusion, although attention was disproportionately directed towards negative information in both disorders, cognitive inhibition deficits appeared specific to OCD. PMID- 15896288 TI - Pain catastrophizing mediates the relationship between worry and pain suffering in patients with irritable bowel syndrome. AB - Although separate lines of behaviorally oriented pain research have drawn attention to the importance of pain catastrophizing and trait worry, little is known about how they work together to influence aspects of chronic pain. Integrating pain research with the broader anxiety, cognitive science, and learning literature, we hypothesized that the process (vs. content) of worry influences pain through catastrophizing. One hundred and eighty-six consecutive patients diagnosed (Rome II) with irritable bowel syndrome completed measures of three dimensions of pain (sensory pain, affective pain, long-term suffering), pain intensity, trait anxiety, worry, catastrophizing, and somatization during baseline assessment of an NIH-funded clinical trial of two psychological treatments. Worry was most strongly associated with the emotionally unpleasant aspects of pain, particularly suffering. Multivariate mediational analyses showed that catastrophizing mediated the link between worry and suffering. Worry, catastrophizing and control variables accounted for 46% of the variance in suffering. Chronic pain patients who worry excessively engage in more catastrophic thinking and through this cognitive process experience more intensely the suffering component of pain. Data are consistent with the notion that worry functions as an "experiential avoidance" strategy for aversive features of pain. Findings are discussed with respect to their relevance to behavioral models for understanding and treating anxiety-related chronic pain disorders. PMID- 15896289 TI - Impact of cognitive-behavioral therapy for panic disorder on comorbidity: a controlled investigation. AB - This study examined the effects of cognitive-behavioral therapy (CBT) for principal panic disorder with or without agoraphobia, on comorbidity in 30 individuals (16 female). To test the hypothesis that improvements in co-existing conditions were not due to spontaneous fluctuations across time, patients receiving immediate CBT were compared to those assigned to wait list (n = 11). Results indicated clinician-rated severity of comorbid specific phobia declined significantly following immediate CBT compared to no change after wait list. The number of patients without comorbidity of any severity increased after immediate CBT, with no such increase following wait list. However, the groups did not differ in the frequency of additional diagnoses or overall severity of comorbidity. In the total sample, results indicated reductions in comorbidity by 9-month follow-up, with marked declines in the severity of comorbid generalized anxiety disorder (GAD), social and specific phobia. Our findings suggest that targeted CBT for panic disorder has beneficial effects on comorbidity over the longer term and that some of its immediate effects exceed those due to the passage of time alone. PMID- 15896290 TI - Is NF-kappaB a culprit in type 2 diabetes? AB - It has been generally viewed that salicylates ameliorate type 2 diabetes through interfering with the NF-kappaB signaling. Earlier studies indicated that IKKbeta was the key for the development of insulin resistance. However, it was unknown whether IKKbeta itself, or its downstream target, NF-kappaB, plays major roles in insulin resistance. New data suggest that NF-kappaB and NF-kappaB-regulated cytokines are crucial for the diabetogenesis. PMID- 15896291 TI - A new aldehyde oxidase selectively expressed in chemosensory organs of insects. AB - Signal termination is a crucial step in the dynamic of the olfactory process. It involves different classes of odorant-degrading enzymes. Whereas aldehyde oxidase enzymatic activities have been demonstrated in insect antennae by previous biochemical studies, the corresponding enzymes have never been characterized at the molecular level. In the cabbage armyworm Mamestra brassicae, we isolated for the first time an aldehyde oxidase partial cDNA specifically expressed in chemosensory organs, with the strongest expression in antennae of both sexes. In these organs, expression was restricted to the olfactory sensilla. Our results suggest that the corresponding enzyme could degrade aldehyde odorant compounds, such as pheromones or plant's volatiles. PMID- 15896292 TI - Adoptive transfer of macrophages ameliorates renal fibrosis in mice. AB - We performed adoptive transfer of bone marrow-derived (BM) macrophages following pharmacological depletion of leukocytes in a mouse model of unilateral ureteral obstruction (UUO). Treatment with cyclophosphamide (CPM) caused marked decrease in the numbers of F4/80-positive interstitial macrophages as well as in peripheral blood leukocyte counts, and adoptive transfer of BM macrophages to CPM treated mice resulted in significant increase in the numbers of interstitial macrophages both at day 5 and at day 14 after UUO. At day 5 after UUO, no significant change was observed in the degree of renal interstitial fibrosis either by treatment with CPM or with CPM+macrophage. However, at day 14 after UUO, treatment with CPM caused significant increase in the degree of interstitial fibrosis, and adoptive macrophage transfer to these mice attenuated this enhancement in renal fibrosis. Our result suggests the role of infiltrating macrophages on facilitating tissue repair at late stage of UUO. PMID- 15896293 TI - Identification and characterization of splice variants of the human P2X7 ATP channel. AB - The P2X7 channel is a member of the P2X family of ligand-gated ion channels which respond to ATP as the endogenous agonist. Studies suggest that P2X7 has a potentially pivotal role in inflammatory responses largely stemming from its role in mediating the release of IL-1beta in response to ATP. We report the identification of seven variants of human P2X7 which result from alternative splicing. Two of these variants (one lacking the first transmembrane domain, the second lacking the entire cytoplasmic tail) were compared to the full-length channel. Real-time PCR analysis demonstrated that both variants were expressed in various tissues and that the cytoplasmic tail deleted variant is highly expressed. Deletion of the first transmembrane domain resulted in a non functional channel. Deletion of the cytoplasmic tail did not affect ion movement but severely affected the ability to form a large pore and to induce activation of caspases. PMID- 15896294 TI - Decrease in glucose-stimulated insulin secretion following exposure to magnetic fields. AB - We evaluated the effects of extremely low frequency magnetic field (ELFMF) on glucose-stimulated insulin secretion from HIT-T15 cells and investigated the mechanisms of these effects. We demonstrated that exposure to ELFMF at 5mT decreased glucose-stimulated insulin secretion by preventing the increases in cellular adenosine 5'-triphosphate/adenosine 5'-diphosphate, membrane depolarization, and cytosolic free calcium ion concentration. The glucose-induced upregulation of insulin mRNA expression was also attenuated by exposure to ELFMF, although cell viability was not affected. These findings demonstrate the potential of exposure to ELFMF for clinical use as a novel inhibitory method of insulin secretion. PMID- 15896295 TI - The conserved WRPW motif of Hes6 mediates proteasomal degradation. AB - Hes6 belongs to a subfamily of basic helix-loop-helix transcription factors that includes Drosophila Hairy and Enhancer of split genes. Like other members of the family, Hes6 features the WRPW motif which is consisted just of four amino acids at its C-terminus. Here, we show that WRPW motif deletion mutant protein is substantially stabilized in comparison to the full length protein and that the enhanced stability is due to its resistance to proteasomal degradation. The WRPW motif also appears to be sufficient for acceleration of proteolysis as its fusion to two heterologous proteins, the green fluorescent protein (GFP) of Aequoria victoria and Gal4 DNA binding domain of Saccharomyces cerevisiae, significantly destabilized the proteins. These findings demonstrate a novel function of this conserved motif as a degradation signal and raise the possibility of utilizing it for controlling the level of ectopically expressed gene products. PMID- 15896296 TI - Ethanol alters cellular activation and CD14 partitioning in lipid rafts. AB - Alcohol consumption interferes with innate immunity. In vivo EtOH administration suppresses cytokine responses induced through Toll-like receptor 4 (TLR4) and inhibits TLR4 signaling. Actually, EtOH exhibits a generalized suppressive effect on signaling and cytokine responses induced by through most TLRs. However, the underlying mechanism remains unknown. RAW264.7 cells were treated with LPS or co treated with EtOH or with lipid raft-disrupting drugs. TNF-alpha production, IRAK 1 activation, and CD14 partition were evaluated. EtOH or nystatin, a lipid raft disrupting drug, suppressed LPS-induced production of TNF-alpha. The suppressive effect of EtOH on LPS-induced TNF-alpha production was additive with that of methyl-beta-cyclodextrin (MCD), another lipid raft-disrupting drug. EtOH interfered with IRAK-1 activation, an early TLR4 intracellular signaling event. Cell fractionation analyses show that acute EtOH altered LPS-related partition of CD14, a critical component of the LPS receptor complex. These results suggest a novel mechanism of EtOH action that involves interference with lipid raft clustering induced by LPS. This membrane action of EtOH might be one of the mechanisms by which EtOH acts as a generalized suppressor for TLR signaling. PMID- 15896297 TI - Mitochondrial DNA deletions sensitize cells to apoptosis at low heteroplasmy levels. AB - A heterogeneous group of multisystem disorders affecting various tissues and often including neuromuscular symptoms is caused by mutations of the mitochondrial genome, which codes 13 polypeptides of oxidative phosphorylation (OXPHOS) complexes and 22 tRNA genes needed for their translation. Since the link between OXPHOS dysfunction and clinical phenotype remains enigmatic in many diseases, a possible role of enhanced apoptosis is discussed besides bioenergetic crisis of affected cells. We analyzed the proapoptotic impact of the mitochondrial 5kb common deletion (CD), affecting five tRNA genes, in transmitochondrial cybrid cell lines and found a slightly enhanced sensitivity to exogenous oxidative stress (H2O2) and a pronounced sensitization against death receptor stimulation (TRAIL) at a rather low CD heteroplasmy level of 22%. Mitochondrial deletions confer enhanced susceptibility against proapoptotic signals to proliferating cells, which might explain the elimination of deletions from hematopoietic stem cells. PMID- 15896298 TI - In cerebrospinal fluid ER chaperones ERp57 and calreticulin bind beta-amyloid. AB - The beta-amyloids (abetas) are the major components of the plaque observed in the brains of patients with Alzheimer's disease. The conundrum is that although they are produced in everyone during the posttranslational processing in the endoplasmic reticulum (ER) of the amyloid precursor protein (APP), deposits are only observed in the elderly. Our work suggests that normals have a carrier protein(s) keeping them in solution. Based on immunoblotting studies of cerebrospinal fluid (CSF) from normals, we find that the bulk of the abetas are bound to the ER chaperones, ERp57 and calreticulin, suggesting that these may be carrier proteins which prevent aggregation of the abetas and that the deposits are due to faulty ER posttranslational processing of APP with the failure to form this complex. If membrane protein synthesis is similarly affected, it could explain the neuronal dysfunction characteristic of Alzheimer's disease. PMID- 15896299 TI - The uncovering of a novel regulatory mechanism for PLD2: formation of a ternary complex with protein tyrosine phosphatase PTP1B and growth factor receptor-bound protein GRB2. AB - The regulation of PLD2 activation is poorly understood at present. Transient transfection of COS-7 with a mycPLD2 construct results in elevated levels of PLD2 enzymatic activity and tyrosyl phosphorylation. To investigate whether this phosphorylation affects PLD2 enzymatic activity, anti-myc immunoprecipitates were treated with recombinant protein tyrosine phosphatase PTP1B. Surprisingly, lipase activity and PY levels both increased over a range of PTP1B concentrations. These increases occurred in parallel to a measurable PTP1B-associated phosphatase activity. Inhibitor studies demonstrated that an EGF-receptor type kinase is involved in phosphorylation. In a COS-7 cell line created in the laboratory that stably expressed myc-PLD2, PTP1B induced a robust (>6-fold) augmentation of myc PLD2 phosphotyrosine content. The addition of growth factor receptor-bound protein 2 (Grb2) to cell extracts also elevated PY levels of myc-PLD (>10-fold). Systematic co-immunoprecipitation-immunoblotting experiments pointed at a physical association between PLD2, Grb2, and PTP1B in both physiological conditions and in overexpressed cells. This is the first report of a demonstration of the mammalian isoform PLD2 existing in a ternary complex with a protein tyrosine phosphatase, PTP1b, and the docking protein Grb2 which greatly enhances tyrosyl phosphorylation of the lipase. PMID- 15896300 TI - Mechanism of PDX-1 protein transduction. AB - PDX-1 plays a central role in differentiation of insulin-producing cells. We previously reported that exogenous PDX-1 protein can permeate cells and induce insulin gene expression in progenitor cells. These data suggest a strategy for facilitating differentiation into insulin-producing cells. Here we show the mechanism of PDX-1 protein transduction. Initially, a punctate cytoplasmic distribution of PDX-1 protein transduction domain (PTD), which co-localized with an endosomal marker, was observed in treated cells. However, homogeneous distribution of PDX-1-PTD was observed in some cells, indicating that PDX-1 is transduced by endocytosis and then released. The experiments using inhibitors suggested that the PDX-1 is transported through the Golgi complex and to the endoplasmic reticulum. Moreover, we observed in real-time PDX-1-PTD release from endosomes. These data suggest that mechanism of transduction of PDX-1 protein is by endocytosis and subsequent release from the endosome homogeneously in cytoplasm and nuclei, and that PDX-1 protein transduction could be a valuable strategy for facilitating differentiation of progenitor cells into insulin producing cells. PMID- 15896301 TI - Activation of phosphatidylinositol 3-kinase by cellular prion protein and its role in cell survival. AB - The cellular prion protein (PrP(C)) is thought to be involved in protection against cell death, however the exact cellular mechanisms involved are still controversial. Herein we present data that strongly indicate a functional link between PrP(C) expression and phosphatidylinositol 3-kinase (PI 3-kinase) activation, a protein kinase that plays a pivotal role in cell survival. Both mouse neuroblastoma N2a cells and immortalized murine hippocampal neuronal cell lines expressing wild-type PrP(C) had significantly higher PI 3-kinase activity levels than their respective controls. Moreover, PI 3-kinase activity was found to be elevated in brain lysates from wild-type mice, as compared to prion protein knockout mice. Recruitment of PI 3-kinase by PrP(C) was shown to contribute to cellular survival toward oxidative stress by using 3-morpholinosydnonimine (SIN 1) and serum deprivation. Moreover, both PI 3-kinase activation and cytoprotection by PrP(C) appeared to rely on copper binding to the N-terminal octapeptide of PrP(C). Thus, we propose a model in which the interaction of copper(II) with the N-terminal domain of PrP(C) enables transduction of a signal to PI 3-kinase; the latter, in turn, mediates downstream regulation of cell survival. PMID- 15896302 TI - Downregulation of ubiquitin-dependent protein degradation in murine myotubes during hyperthermia by eicosapentaenoic acid. AB - Muscle atrophy in a number of acute wasting conditions is associated with an increased activity and expression of the ubiquitin-proteasome proteolytic pathway. Although different initiators are involved, it is possible that the intracellular signalling events leading to upregulation of this pathway are the same in all catabolic conditions. This study investigates hyperthermia in murine myotubes as a model for increased protein degradation through the ubiquitin proteasome pathway. The effect of eicosapentaenoic acid (EPA) on this process should identify common elements, since EPA has been shown to attenuate induction of the ubiquitin-proteasome pathway in cancer cachexia. Increasing the temperature of myotubes caused a progressive increase in protein degradation. This was associated with an increased proteasome 'chymotrypsin-like' enzyme activity, as well as increased expression of both mRNA and protein for 20S proteasome subunits and the ubiquitin-conjugating enzyme (E2(14k)). This upregulation was not seen in cultures treated with EPA (50 microM), suggesting that it acts to prevent transcriptional activation of the ubiquitin-proteasome pathway in hyperthermia. These results suggest that protein catabolism in hyperthermia and cancer cachexia is mediated through a common pathway. PMID- 15896303 TI - Involvement of MEK-ERK signaling pathway in the inhibition of cell growth by troglitazone in human pancreatic cancer cells. AB - In the present study, we examined a role of mitogen-activated protein kinases (MAPKs), extracellular signal-related kinase (ERK), c-Jun N-terminal protein kinase, and p38 MAPK in troglitazone-induced inhibition of cell growth in human pancreatic cancer cells. Among the three kinases, troglitazone specifically inhibited the phosphorylation of ERK1/2 in a dose- and time-dependent manner. Troglitazone also down-regulated the protein expression of mitogen-activated protein kinase kinase (MEK)1/2, an upstream molecule that regulates ERK phosphorylation. Treatment of human pancreatic cancer cells with specific MEK inhibitor, PD98059 or U0126, inhibited ERK1/2 phosphorylation and cell growth. These results suggest for the first time that the inhibition of the MEK1/2-ERK1/2 signaling pathway may be implicated in the growth inhibitory effect by troglitazone in human pancreatic cancer cells. PMID- 15896304 TI - Genotype of inflammatory cytokines in limbal stem cell graft in Italian patients. AB - We tested the hypothesis that the genetic capability to mount an inflammatory response might contribute to the inter-individual variability of limbal stem cell graft (LSCG) outcome. Two functional polymorphisms in the IL-6 and TNF-alpha promoter regions were genotyped in 35 patients. A new score system (clinical assessment score, CAS) was set up in order to classify patients' clinical profile, and the main parameters relevant for LSCG as well as for the follow-up of the patients. Patients carrying at both loci a genotype associated with a lower production of both cytokines were classified as "low producers" (LP), while all the others were classified as "intermediate or high producers" (HP). LP patients did not show any difference in CAS before and after transplantation while a significant difference was present in HP patients. A similar trend was evident in the 35 months of follow-up. Polymorphisms of IL-6 and TNF-alpha can be used to identify subgroups of patients with higher risk of unsuccessful outcome. PMID- 15896305 TI - Differential regulation of diacylglycerol kinase isozymes in cardiac hypertrophy. AB - To examine the involvement of diacylglycerol kinase (DGK) and phosphatidic acid phosphatase (PAP) in pressure overloaded cardiac hypertrophy, rats were subjected to either ascending aortic banding for 3, 7, and 28 days or sham operation. In comparison with sham-operated rats, the left ventricular (LV) weight of the aortic-banded rats increased progressively. At 28 days after surgery, the expression of DGKepsilon mRNA but not DGKzeta or PAP2b mRNA in the LV myocardium significantly decreased in the aortic-banded rats compared with the sham-operated rats. DGKzeta protein in the LV myocardium translocated from the particulate to the cytosolic compartment in the aortic-banded rats. Furthermore, the myocardial content of 1,2-diacylglycerol and PKCdelta protein expression in the particulate fraction of the LV myocardium significantly increased in aortic-banded rats compared with sham-operated rats. These results suggest that DGKepsilon and DGKzeta play distinct roles in the development of pressure overloaded cardiac hypertrophy and that the two isozymes are differentially regulated. PMID- 15896306 TI - Detection of nanometer-sized particles in living cells using modern fluorescence fluctuation methods. AB - Nanosized materials are increasingly used in medicine and biotechnology but originate also from various aerosol sources. A detailed understanding of their interaction with cells is a prerequisite for specific applications and appraisal of hazardous effects. Fluorescence fluctuation methods are applied to follow the time-course of the translocation and distribution of fluorescent 20 nm polystyrene nanoparticles with negative surface charges in HeLa cells under almost physiological conditions. The experimental results demonstrate that singular particles enter the cell without significant contribution by endocytotic mechanisms and are distributed within the cytoplasm. Subsequently aggregation is observed, which can be blocked by cytotoxins, like Genistein and Cytochalasin B, interfering with cellular uptake processes. The observed non-active uptake is due to non-specific interactions with the cell surface and could be responsible for distribution of nanometer-sized materials in tissue. PMID- 15896307 TI - The CXCR1 tail mediates beta1 integrin-dependent cell migration via MAP kinase signaling. AB - In this study, we examined how IL-8 induces leukocyte migration on major beta1 integrin ligands derived from the extracellular matrix protein fibronectin. We assessed individual contributions of signaling by IL-8 receptors by transfection of CXCR1 and CXCR2 into rat basophilic leukemia (RBL) cells and human monocytic THP-1 cells. CXCR1 expressing cells migrated on the fibronectin ligands for alpha4beta1 and alpha5beta1 integrins in response to IL-8, whereas CXCR2 expressing cells did not. RBL cells expressing the chimeric CXCR1 receptor containing the cytoplasmic tail of CXCR2 had greatly blunted migration, while cells expressing the CXCR2 chimera with the tail of CXCR1 had augmented migration. Last, inhibitors of p38 and JNK MAP kinases blocked IL-8-induced migration in CXCR1+ cells. We conclude that IL-8 stimulated beta1 integrin mediated leukocyte migration on fibronectin through CXCR1 is dependent on the C terminal cytoplasmic domain of CXCR1 and subsequent p38 and JNK MAPK signaling. PMID- 15896308 TI - Expression of Reg/PAP family members during motor nerve regeneration in rat. AB - In this study, we examined the expression of mRNAs for Regenerating gene (Reg)/pancreatitis-associated protein (PAP) family members following hypoglossal nerve injury in rats. In addition to four rat family members (RegI, Reg-2/PAP I, PAP II, and PAP III) that had been identified, we newly cloned and sequenced a type-IV Reg gene in rats. Among these five family members, the expression of Reg 2/PAP I mRNA was predominantly enhanced in injured motor neurons after axotomy. Furthermore, a marked induction of PAP III mRNA was observed in the distal part of the injured nerve. A polyclonal antibody was raised against PAP III, and a Western blotting analysis using this antibody confirmed an increased level of PAP III protein in the injured nerve. These results suggest that Reg family members would be new mediators among injured neurons and glial cells, and may play pivotal roles during nerve regeneration. PMID- 15896309 TI - Transforming growth factor-beta2 enhances differentiation of cardiac myocytes from embryonic stem cells. AB - Stem cell therapy holds great promise for the treatment of injured myocardium, but is challenged by a limited supply of appropriate cells. Three different isoforms of transforming growth factor-beta (TGF-beta) -beta1, -beta2, and -beta3 exhibit distinct regulatory effects on cell growth, differentiation, and migration during embryonic development. We compared the effects of these three different isoforms on cardiomyocyte differentiation from embryonic stem (ES) cells. In contrast to TGF-beta1, or -beta3, treatment of mouse ES cells with TGF beta2 isoform significantly increased embryoid body (EB) proliferation as well as the extent of the EB outgrowth that beat rhythmically. At 17 days, 49% of the EBs treated with TGF-beta2 exhibited spontaneous beating compared with 15% in controls. Cardiac myocyte specific protein markers sarcomeric myosin and alpha actin were demonstrated in beating EBs and cells isolated from EBs. In conclusion, TGF-beta2 but not TGF-beta1, or -beta3 promotes cardiac myocyte differentiation from ES cells. PMID- 15896310 TI - Regional determinants of arterial endothelial phenotype dominate the impact of gender or short-term exposure to a high-fat diet. AB - Regional arterial hemodynamics correlates with distinct endothelial phenotypes that may be modified by risk factors to influence focal and regional susceptibility to atherosclerosis. We compared endothelial transcript profiles from hemodynamically distinct arterial regions in 15 mature pigs: males and females fed a normal diet, and males fed a high-fat diet (15% lard, 1.5% cholesterol) for two weeks. Hierarchical clustering analysis showed preferential grouping of arrays by region over risk factor. A set of differentially expressed genes was identified which clearly distinguished regions of disturbed flow from undisturbed flow; however, few differences were observed within the same region based on gender or diet. Consistent with previous results in the absence of risk factors, the balance in gene expression was not inherently pathological at this early time-point. The results implicate regional hemodynamics as a predominant epigenetic determinant of endothelial phenotypic heterogeneity underlying atherosusceptibility in vivo. PMID- 15896311 TI - Two membrane juxtaposed signaling modules in ANF-RGC are interlocked. AB - Atrial natriuretic factor (ANF) receptor guanylate cyclase ANF-RGC is a single transmembrane spanning modular protein. Juxtaposed to each side of the transmembrane module is a Cys423-Cys432 disulfide ANF signaling module motif and the ATP-regulated transduction module (ARM) motif. The signaling module motif is conserved in nearly all membrane guanylate cyclases and is believed to be critical in the signaling activities of all membrane guanylate cyclases. The present study with the model system of the olfactory membrane guanylate cyclase shows that this concept is not valid. Furthermore, the study shows that in ANF-GC the signaling motif works through the ARM domain. A new signaling model is proposed where in its natural state the disulfide structural motif represses the ARM domain activity, which, in turn, represses the catalytic module activity of ANF-RGC. ANF signaling relieves the disulfide structural motif restraint on the ARM inhibition and stimulates the catalytic module of the cyclase. PMID- 15896312 TI - An N-terminal nuclear localization sequence but not the calmodulin-binding domain mediates nuclear localization of nucleomorphin, a protein that regulates nuclear number in Dictyostelium. AB - Nucleomorphin is a novel nuclear calmodulin (CaM)-binding protein (CaMBP) containing an extensive DEED (glu/asp repeat) domain that regulates nuclear number. GFP-constructs of the 38 kDa NumA1 isoform localize as intranuclear patches adjacent to the inner nuclear membrane. The translocation of CaMBPs into nuclei has previously been shown by others to be mediated by both classic nuclear localization sequences (NLSs) and CaM-binding domains (CaMBDs). Here we show that NumA1 possesses a CaMBD (171EDVSRFIKGKLLQKQQKIYKDLERF195) containing both calcium dependent-binding motifs and an IQ-like motif for calcium-independent binding. GFP-constructs containing only NumA1 residues 1-129, lacking the DEED and CaMBDs, still localized as patches at the internal periphery of nuclei thus ruling out a direct role for the CaMBD in nuclear import. These constructs contained the amino acid residues 48KKSYQDPEIIAHSRPRK64 that include both a putative bipartite and classical NLS. GFP-bipartite NLS constructs localized uniformly within nuclei but not as patches. As with previous work, removal of the DEED domain resulted in highly multinucleate cells. However as shown here, multinuclearity only occurred when the NLS was present allowing the protein to enter nuclei. Site-directed mutation analysis in which the NLS was changed to 48EF49 abolished the stability of the GFP fusion at the protein but not RNA level preventing subcellular analyses. Cells transfected with the 48EF49 construct exhibited slowed growth when compared to parental AX3 cells and other GFP-NumA1 deletion mutants. In addition to identifying an NLS that is sufficient for nuclear translocation of nucleomorphin and ruling out CaM-binding in this event, this work shows that the nuclear localization of NumA1 is crucial to its ability to regulate nuclear number in Dictyostelium. PMID- 15896313 TI - Akt1 contains a functional leucine-rich nuclear export sequence. AB - Nuclear Akt1 expression and Akt activation are common in cancer invasion. However, the mechanisms for this association and its causal role in invasion are uncertain. In an effort to identify potential mechanisms for regulating Akt subcellular localization, we analyzed the Akt gene sequences and identified a highly conserved leucine-rich potential nuclear export sequence (NES). Initial experiments demonstrated that leptomycin B induced nuclear Akt1 localization. Transient expression experiments demonstrated that, in comparison to wild-type Akt1, NES-mutated (AKT/NES) Akt1 has reduced interactions with CRM-1 and persistent nuclear localization. Subsequent stable transfection experiments in Akt1-/- fibroblasts confirmed that expression of AKT/NES resulted in persistent nuclear localization and activation1. Finally, stable expression of AKT/NES in Akt1-/- fibroblasts was sufficient to enhance cell migration in vitro. Thus, Akt1 contains a functional NES and mutation of the NES results in nuclear-predominant Akt1 activation that is sufficient to induce migration. PMID- 15896314 TI - Posttranslational processing of SREBP-1 in rat hepatocytes is regulated by insulin and cAMP. AB - Insulin and cAMP have opposing effects on de novo fatty acid synthesis in liver and in cultured hepatocytes mediated by sterol-regulatory element binding protein (SREBP). To determine whether these agents regulate the cleavage of full-length SREBP to generate the transcriptionally active N-terminal fragment (nSREBP) in primary rat hepatocytes, an adenoviral vector (Ad-SREBP-1a) was constructed to constitutively express full-length SREBP-1a. Insulin increased, and dibutyryl (db)-cAMP inhibited, generation of nSREBP-1a from its full-length precursor. Insulin stimulated processing of SREBP-1a within 1h, and the effect was sustained for at least 24h. The initial stimulation of SREBP processing by insulin preceded measurable reduction in Insig-2 mRNA levels. Rat hepatocytes were also infected with an adenovirus expressing the nuclear form of SREBP-1c (Ad-nSREBP-1c). Insulin increased the half-life of constitutively expressed nSREBP-1c, and this effect of insulin was also inhibited by db-cAMP. PMID- 15896315 TI - RIP3 beta and RIP3 gamma, two novel splice variants of receptor-interacting protein 3 (RIP3), downregulate RIP3-induced apoptosis. AB - Receptor-interacting protein 3 (RIP3) is an apoptosis inducing member of the RIP family. Here we report two novel splice variants of human RIP3, designated RIP3 beta and RIP3 gamma respectively. Unlike full-length RIP3, both variants possess a truncated N-terminal kinase domain and a distinct and shorter C terminus, and therefore abrogate nucleocytoplasmic shuttling and apoptosis-inducing activity. Transient expression of either variant was found to downregulate RIP3-mediated apoptosis. Importantly, real-time PCR analysis reveals that the ratio of RIP3 gamma to RIP3 is significantly increased in colon and lung cancers relative to their matched normal tissues, indicating that RIP3 gamma might be a major splice form associated with tumorigenesis. PMID- 15896316 TI - The mouse CCR2 gene is regulated by two promoters that are responsive to plasma cholesterol and peroxisome proliferator-activated receptor gamma ligands. AB - We have previously shown that the expression of monocyte CCR2, the receptor for monocyte chemoattractant protein-1, is induced by plasma cholesterol. The present study examines the mechanisms that regulate monocyte CCR2 expression in hypercholesterolemia using a mouse model. Our data demonstrate that in the mouse, CCR2 expression in circulating monocytes is controlled by two promoters P1 and P2. The two distinct transcripts, which encode the same protein, are produced by alternative splicing in the 5'-untranslated region. Both promoters are constitutively active, but only P2 is stimulated by cholesterol. However, both promoters are repressed by peroxisome proliferator-activated receptor gamma. PMID- 15896317 TI - Systemic soluble Tie2 expression inhibits and regresses corneal neovascularization. AB - This study was designed to determine if soluble Tie2 (sTie2) expression inhibits and regresses corneal neovascularization, and if VEGF contributes to its effect. The corneas of BALB/c mice were scraped and the mice were injected with either an adenovirus expressing soluble Tie2 (Ad.sTie2) or an empty adenoviral vector. When injected at the inhibition timepoint (one day prior to corneal injury), the mean percentage of neovascularized corneal area two weeks later in Ad.sTie2-treated mice vs. controls was 56.37+/-9.15% vs. 85.79+/-3.55% (p=0.04). At the regression timepoint (4 weeks after corneal scrape), the mean area of corneal neovascularization in Ad.sTie2-treated mice was 42.89+/-4.74% vs. 75.01+/-3.22% in the control group (p=0.007). VEGF expression was significantly higher in Ad.sTie2-treated mice at the inhibition timepoint and there was no significant difference at the regression timepoint. These findings suggest that sTie2 inhibits and regresses corneal neovascularization in a VEGF-independent manner. PMID- 15896318 TI - Stimulated HSP90 binding to eNOS and activation of the PI3-Akt pathway contribute to globular adiponectin-induced NO production: vasorelaxation in response to globular adiponectin. AB - The present study examined potential interactions between endothelial NO synthase (eNOS), heat shock protein (HSP)90, and Akt in vascular endothelial cells stimulated with globular adiponectin to produce nitric oxide (NO). Globular adiponectin-induced eNOS phosphorylation was accompanied by eNOS-HSP90-Akt complex formation, resulting in a dose-dependent increase in NO release. Globular adiponectin stimulated binding of HSP90 to eNOS, and inhibition of HSP90 significantly suppressed globular adiponectin-stimulated NO release. Globular adiponectin also caused Akt phosphorylation, and inhibition of PI3 kinase significantly suppressed globular adiponectin-stimulated NO release. This study also examined whether globular adiponectin really induces endothelial-dependent vasodilation using rings from rat thoracic aorta. It was observed that globular adiponectin caused dose-dependent vasorelaxation in the aorta. These results indicate that stimulated HSP90 binding to eNOS and activation of the PI3-Akt pathway contribute to globular adiponectin-induced eNOS phosphorylation and NO production, and to endothelium-dependent vasorelaxation. PMID- 15896319 TI - Expression of the CDR1 efflux pump in clinical Candida albicans isolates is controlled by a negative regulatory element. AB - Resistance to azole antifungal drugs in clinical isolates of the human fungal pathogen Candida albicans is often caused by constitutive overexpression of the CDR1 gene, which encodes a multidrug efflux pump of the ABC transporter superfamily. To understand the relevance of a recently identified negative regulatory element (NRE) in the CDR1 promoter for the control of CDR1 expression in the clinical scenario, we investigated the effect of mutation or deletion of the NRE on CDR1 expression in two matched pairs of azole-sensitive and resistant clinical isolates of C. albicans. Expression of GFP or lacZ reporter genes from the wild type CDR1 promoter was much higher in the azole-resistant C. albicans isolates than in the azole-susceptible isolates, reflecting the known differences in CDR1 expression in these strains. Deletion or mutation of the NRE resulted in enhanced reporter gene expression in azole-sensitive strains, but did not further increase the already high CDR1 promoter activity in the azole-resistant strains. In agreement with these findings, electrophoretic mobility shift assays showed a reduced binding to the NRE of nuclear extracts from the resistant C. albicans isolates as compared with extracts from the sensitive isolates. These results demonstrate that the NRE is involved in maintaining CDR1 expression at basal levels and that this repression is overcome in azole-resistant clinical C. albicans isolates, resulting in constitutive CDR1 overexpression and concomitant drug resistance. PMID- 15896320 TI - Effect of neurturin on multipotent cells isolated from the adult skeletal muscle. AB - Ligands of the glial cell line-derived neurotrophic factors (GDNF)-family are trophic factors for the development and survival of multiple cell types, however their effects on non-neuronal stem cells are unknown. We examined the action of neurturin on a candidate stem cell population isolated from adult skeletal muscles. When grown as spheres, these cells expressed mRNAs for GDNF, persephin, GFR-alpha2, GFR-alpha4 (neurturin receptor), and Ret. Exposure of these cells to neurturin significantly augmented cell numbers via increased cell proliferation. After addition of retinoic acid, the cells exited the cell cycle, developed thin processes, and became immunoreactive for betaIII-tubulin, while Ret mRNA expression decreased, without changes in the level of GFR-alpha2 mRNA. Neurturin induced an outgrowth of processes on these betaIII-tubulin positive cells. Neurturin may therefore be beneficial in the use of these multipotent cells isolated from adult muscles for autologous transplants in neurological applications. PMID- 15896321 TI - Two L1-peptides are excellent tools for serological detection of HPV-associated cervical carcinoma lesions. AB - A persistent high risk human papillomavirus (HR-HPV) infection causes cervical intraepithelial lesions and cervical carcinoma. There is evidence that detecting anti-L1 antibodies could be successfully used for discriminating between cervical lesion patients and women having normal cytology. It was found that peptides 18283 (55PNNNKILVPKVSGLQYRVFR74) and 18294 (284LYIKGSGSTANLASSNYFPT300) from the L1-surface exposed regions were specifically recognised by antibodies from the cervical lesion patient sera. These peptides were tested against 165 womens' normal cytology sera and 148 cervical lesion or cervical cancer patients' sera. Less than 3.6% of women's normal cytology sera recognised peptides 18283 or 18294; on the contrary, 91% to 96% of the cervical lesion (CIN I to CIN III) or cervical cancer patient sera recognised peptides 18283 and 18294. These data show that anti-peptide 18283 and 18294 antibodies in the patients' sera are strongly associated with the presence of HR-HPV associated cervical lesions, showing 92 97% sensitivity and 89-95% specificity in recognising precancerous and cervical cancer patients. These two peptides could be excellent tools for use in large scale serological screening of women populations at risk of developing cervical carcinoma. PMID- 15896322 TI - Common anti-apoptotic roles of parkin and alpha-synuclein in human dopaminergic cells. AB - Parkin, a product of the gene responsible for autosomal recessive juvenile parkinsonism (AR-JP), is an important player in the pathogenic process of Parkinson's disease (PD). Despite numerous studies including search for the substrate of parkin as an E3 ubiquitin-protein ligase, the mechanism by which loss-of-function of parkin induces selective dopaminergic neuronal death remains unclear. Related to this issue, here we show that antisense knockdown of parkin causes apoptotic cell death of human dopaminergic SH-SY5Y cells associated with caspase activation and accompanied by accumulation of oxidative dopamine (DA) metabolites due to auto-oxidation of DOPA and DA. Forced expression of alpha synuclein (alpha-SN), another familial PD gene product, prevented accumulation of oxidative DOPA/DA metabolites and cell death caused by parkin loss. Our findings indicate that both parkin and alpha-SN share a common pathway in DA metabolism whose abnormality leads to accumulation of oxidative DA metabolites and subsequent cell death. PMID- 15896323 TI - Photoreceptor cells from mouse ES cells by co-culture with chick embryonic retina. AB - Degeneration of photoreceptors is a consistent and common endpoint in retinal diseases. Herein, we report the efficient induction of photoreceptor-like cells from mouse embryonic stem (ES) cells using chick embryonic retina tissue. Undifferentiated mouse ES cells were initially cultured in a preferential condition into a neural lineage, and ES cells were then co-cultured with chick embryonic day 6 (E6) retina tissues. After a 10-day co-culture, approximately 20% of the mouse ES derivatives became immuno-positive for rhodopsin. RT-PCR analysis demonstrated expression of the transcription factor crx and a distinct increase of rod photoreceptor-specific markers, IRBP and recoverin, after the start of the co-culture. These results indicate that co-culture of ES cells with chick embryonic retina tissue is a useful and efficient method for the induction of photoreceptor-like cells. PMID- 15896324 TI - Altered cathepsin D metabolism in PHEX antisense human osteoblast cells. AB - X-linked hypophosphatemia (XLH), the most common form of hereditary rickets, is caused by loss-of-function mutations of PHEX gene in osteoblast cells, leading to rachitic bone disease and hypophosphatemia. Available evidence today indicates that the bone defect in XLH is caused not only by hypophosphatemia and altered vitamin D metabolism, but also by locally released osteoblastic mineralization inhibitory factor(s), referred to as minhibin. In our present study, we found that suppression of PHEX expression by PHEX antisense in human osteoblast cells caused an increase in cathepsin D expression at protein, but not mRNA, levels. This was associated with a decrease in cathepsin D degradation and an increased cathepsin D release into culture media. Our results also showed that lowering cathepsin D activity in antisense cell conditioned media abolished their inhibitory effect on osteoblast cell calcification, suggesting the involvement of cathepsin D in mediating the minhibin activity of the antisense cell conditioned media. PMID- 15896325 TI - Multiple transcripts regulate glucose-triggered mRNA decay of the lactate transporter JEN1 from Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae JEN1 gene encoding the lactate transporter undergoes strong catabolic repression at both transcriptional and post-transcriptional levels. JEN1 mRNA decay is greatly accelerated upon the addition of a pulse of glucose, fructose or mannose to induced cell cultures. Mapping of the 5'UTRs and 3'UTRs of JEN1 transcripts revealed multiple transcription start-sites located at position -51, +391 or +972, depending on the cell culture conditions. The presence of the JEN1(+391) transcript correlated with rapid glucose-triggered mRNA degradation of the JEN1(-51) transcript, whereas when the small transcript started at position +972, the JEN1(-51) mRNA turnover rate was unaffected. Overexpressed JEN1(+391) transcript accelerated JEN1(-51) mRNA decay in all conditions tested but was not translated. We propose that the JEN1(+391) transcript may have a "sensor-like" function, regulating glucose-triggered degradation of JEN1(-51) protein-coding mRNA. PMID- 15896326 TI - Turnover and characterization of UDP-N-acetylglucosaminyl transferase in a stably transfected HeLa cell line. AB - To estimate the turnover of UDP-N-acetylglucosaminyl transferase (OGT), we exposed stably transfected HeLa cells to tetracycline for 16h to induce OGT gene expression and increase cytosolic enzyme levels. Removal of tetracycline led to a progressive decrease in OGT activity (after a 6h lag period), yielding an estimated OGT half-life of 13h. A similar half-life (12h) was obtained by measuring the loss of biosynthetically labeled OGT ([35S]methionine pulse-chase experiments). Since OGT turnover was relatively slow, it is unlikely that changes in OGT gene expression or protein expression play a role in the short-term regulatory actions mediated by the hexosamine signaling pathway. We also found that the overexpressed 110kDa murine OGT subunit (recombinant enzyme) was enzymatically similar to the endogenous holoenzyme derived from rat brain tissue. Thus, stably transfected HeLa cells provide an abundant source of enzyme that can be used to study the structure, function, and regulation of OGT. PMID- 15896327 TI - Nuclear localization of long-VEGF is associated with hypoxia and tumor angiogenesis. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic factor that has a pivotal role in normal and pathological angiogenesis. VEGF has a long 5' untranslated region harboring an open reading frame (ORF) initiated by a CUG codon that is in-frame with the VEGF coding region. The ORF translation leads to the expression of a long isoform termed L-VEGF that is extended by an additional 180 amino acids. In this communication, we provide evidence that L-VEGF is subjected to proteolytic cleavage leading to the detachment of the 180 aa extension from the VEGF moiety. Using immunofluorescence staining, we show that upon hypoxia this 180 aa extension translocates to the nuclei of expressing cells. Accordingly, immunohistochemical staining of both normal and tumor tissue samples demonstrated restricted nuclear localization of the ORF, which was correlated with cytoplasmic localization of VEGF. This suggests that the 180 aa ORF is involved in VEGF-mediated angiogenic processes. PMID- 15896328 TI - Cellular trafficking of the IL-1RI-associated kinase-1 requires intact kinase activity. AB - Upon stimulation of cells with interleukin-1 (IL-1) the IL-1 receptor type I (IL 1RI) associated kinase-1 (IRAK-1) transiently associates to and dissociates from the IL-1RI and thereafter translocates into the nucleus. Here we show that nuclear translocation of IRAK-1 depends on its kinase activity since translocation was not observed in EL-4 cells overexpressing a kinase negative IRAK-1 mutant (EL-4(IRAK-1-K239S)). IRAK-1 itself, an endogenous substrate with an apparent molecular weight of 24kDa (p24), and exogenous substrates like histone and myelin basic protein are phosphorylated by nuclear located IRAK-1. Phosphorylation of p24 cannot be detected in EL-4(IRAK-1-K239S) cells. IL-1 dependent recruitment of IRAK-1 to the IL-1RI and subsequent phosphorylation of IRAK-1 is a prerequisite for nuclear translocation of IRAK-1. It is therefore concluded that intracellular localization of IRAK-1 depends on its kinase activity and that IRAK-1 may also function as a kinase in the nucleus as shown by a new putative endogenous substrate. PMID- 15896329 TI - Transcriptional regulation of mouse 6-phosphogluconate dehydrogenase by ADD1/SREBP1c. AB - 6-Phosphogluconate dehydrogenase (6PGDH) constitutes the pentose phosphate pathway and produces NADPH. 6PGDH is also considered as a lipogenic gene since NADPH is a pivotal cofactor for lipogenesis. Thus, it is important to elucidate how 6PGDH is regulated by various signals related to energy homeostasis. Here, we provide several evidences that ADD1/SREBP1c regulates the expression of mouse 6PGDH gene. DNase I footprinting assay and point mutation studies revealed that the E-box (CANNTG) motif in the promoter of mouse 6PGDH is an important cis regulatory element for ADD1/SREBP1c. 6PGDH mRNA is highly expressed in white adipose tissue and tightly modulated by nutritional status. Furthermore, we found that ADD1/SREBP1c mediates insulin-dependent 6PGDH expression and that PI3-kinase is an important linker for its regulation. Taken together, these data suggest that ADD1/SREBP1c is a key transcription factor for 6PGDH gene expression and would coordinate glucose metabolism and lipogenesis for energy homeostasis. PMID- 15896330 TI - Molecular markers distinguish bone marrow mesenchymal stem cells from fibroblasts. AB - To characterize mesenchymal stem cells (MSC), we compared gene expression profiles in human bone marrow MSC (11 lines) and human fibroblasts (4 lines) by RT-PCR and real time PCR. Messenger RNA levels of MHC-DR-alpha, MHC-DR-beta, MHC DR-associated protein CD74, tissue factor pathway inhibitor-2, and neuroserpin were much higher in MSC than in fibroblasts, even in the presence of large interindividual variations. Those of adrenomedullin, apolipoprotein D, C-type lectin superfamily member-2, collagen type XV alpha1, CUG triplet repeat RNA binding protein, matrix metalloproteinase-1, protein tyrosine kinase-7, and Sam68 like phosphotyrosine protein/T-STAR were lower in MSC than in fibroblasts. FACS analysis showed that cell surface expression of MHC-DR was also higher in MSC than in fibroblasts. MHC-DR expression decreased after osteogenic differentiation, whereas the expression of adrenomedullin-a potent stimulator of osteoblast activity-along with collagen XV alpha1 and apolipoprotein D increased after osteogenic differentiation. The marker genes identified in this study should be useful for characterization of MSC both in basic and clinical studies. PMID- 15896331 TI - Characterization of the cloned full-length and a truncated human target of rapamycin: activity, specificity, and enzyme inhibition as studied by a high capacity assay. AB - The mammalian target of rapamycin (mTOR/TOR) is implicated in cancer and other human disorders and thus an important target for therapeutic intervention. To study human TOR in vitro, we have produced in large scale both the full-length TOR (289 kDa) and a truncated TOR (132 kDa) from HEK293 cells. Both enzymes demonstrated a robust and specific catalytic activity towards the physiological substrate proteins, p70 S6 ribosomal protein kinase 1 (p70S6K1) and eIF4E binding protein 1 (4EBP1), as measured by phosphor-specific antibodies in Western blotting. We developed a high capacity dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) for analysis of kinetic parameters. The Michaelis constant (Km) values of TOR for ATP and the His6-S6K substrate were shown to be 50 and 0.8 microM, respectively. Dose-response and inhibition mechanisms of several known inhibitors, the rapamycin-FKBP12 complex, wortmannin and LY294002, were also studied in DELFIA. Our data indicate that TOR exhibits kinetic features of those shared by traditional serine/threonine kinases and demonstrate the feasibility for TOR enzyme screen in searching for new inhibitors. PMID- 15896332 TI - Beta2-microglobulin required for cell surface expression of blastocyst MHC. AB - Blastocyst MHC is a mouse MHC class Ib gene that is selectively expressed in blastocysts and placenta like human HLA-G, which protect fetal trophoblasts and some tumor cells from NK cell attack, and in TAP-dependent expression on the cell surface. We expressed blastocyst MHC cDNA in beta2-deficient EL-4 S3 or beta2m transfected EL-4 S3 cells. In parental EL-4 S3 cells, only 47-kDa blastocyst MHC protein was expressed and retained in the cytoplasm. However, additional 51-kDa blastocyst MHC protein was expressed on the surface of beta2m-transfected EL-4 S3 cells. The 51-kDa protein was resistant to Endo-H, whereas the 47-kDa protein was sensitive for Endo-H. The results suggested that beta2m as well as TAP was necessary for the transportation of blastocyst MHC from endoplasmic reticulum to cell surfaces through the Golgi apparatus, similar to other MHC class I molecules. PMID- 15896333 TI - Role of PI3K/Akt and MEK/ERK signaling pathways in sulforaphane- and erucin induced phase II enzymes and MRP2 transcription, G2/M arrest and cell death in Caco-2 cells. AB - Isothiocyanate sulforaphane is an extensively studied cancer chemopreventive agent in human diet. In this study, the effects of sulforaphane (SFN) and its sulfide analog, erucin (ERN), have been examined on the induction of the phase II enzymes, quinine oxidoreductase (NQO1) and UDP-glucuronosyl transferase (UGT1A1), multidrug transporter (MRP2), cell cycle arrest and cell death in human colon adenocarcinoma Caco-2 cells. Additionally, the roles of PI3K/Akt and MEK/ERK signaling pathways have been assessed in these sulforaphane- and erucin-induced events. Although erucin and sulforaphane have similar IC50 values (21 and 23 microM after 72 h treatment), erucin was more effective in the induction of G2/M accumulation, depletion of mitochondrial potential, induction of cell death and mRNA induction of phase II enzymes and MRP2. Erucin (20 microM) induced the mRNAs of NQO1, UGT1A1 and MRP2 by 11.1-, 11.6- and 6.7-fold, whereas sulforaphane (20 microM) induced 3.3-, 5.3- and 2.2-fold, respectively. Both erucin and sulforaphane induced activation (phosphorylation) of ERK1/2 and Akt kinases but had no effect on JNK and p38 activation. Erucin-induced phase II enzyme transcriptions were decreased by PI3K and MEK1 inhibitors (LY294002 and PD98059), but the decreases in sulforaphane-induced transcription were less marked. Erucin induced a large increase in G2/M cell number than sulforaphane. The ability of kinase inhibitors to overcome G2/M block was low with the exception of PD98059 in sulforaphane-treated cells. Both, sulforaphane and erucin at high concentrations induced accumulation of sub-G1 cells, cell death and dissipation of mitochondrial membrane potential. Taken together, these results demonstrate that PI3K/Akt and MEK/ERK signals are important intracellular mediators in erucin- and sulforaphane mediated phase II enzyme transcription and cell cycle arrest in Caco-2 cells. PMID- 15896334 TI - The potential benefits of 1.5% hetastarch as a cardioplegia additive. AB - INTRODUCTION: Myocardial edema is a clinically relevant problem found in post ischemic reperfused hearts. The objective of this study was to understand the effects of hetastarch-supplemented cardioplegia on post-ischemic edema and cardiac function. MATERIALS AND METHODS: Swine hearts were arrested with either St. Thomas Hospital cardioplegia with (n=6) or without (n=7) 1.5% hetastarch. Following hypothermic global ischemia, hearts were crystalloid reperfused in a four-chamber isolated working mode. RESULTS: Hetastarch decreased myocardial water content gains after three hours of reperfusion (control versus hetastarch, hour 0: 67+/-5% versus 67+/-3%, NS; hour 3: 82+/-2% versus 78+/-1%, p=0.1). Post ischemic control group left ventricular end-diastolic pressures were elevated after 1h (in mm Hg, hour 0: 13+/-2, hour 1: 19+/-3, hour 2: 19+/-3, hour 3: 20+/ 2) but remained stable (<16 mm Hg) in the hetastarch group. Post-reperfusion creatine phosphokinase perfusate levels in the hetastarch treated hearts were decreased (control: 1.6 IU/l/g versus hetastarch: 0.6 IU/l/g, p=0.15). DISCUSSION/CONCLUSIONS: Hetastarch treatment delayed myocardial edema development and attenuated myocardial creatine kinase efflux, thereby preserving diastolic function. PMID- 15896335 TI - Antisense gene delivered by an adenoassociated viral vector inhibits iron uptake in human intestinal cells: potential application in hemochromatosis. AB - Hereditary hemochromatosis (HH) is a condition in which intestinal iron absorption is greatly elevated. Present treatment is weekly phlebotomy, affecting quality of life and leading to recurrent infections. The iron transporter divalent metal transporter-1 (DMT-1) of enterocytes is responsible for iron uptake from the intestinal lumen; iron is further extruded into the blood by the basolateral transporter ferroportin-1. A therapeutic approach for HH could start with a long-term reduction of iron transport by reduction of DMT-1 levels. We designed an AAV vector coding for a short antisense RNA (AAV-DMT-1-AS) against DMT-1, which reduced iron uptake by 50-60% in human intestinal cells (Caco-2). At low infection levels, DMT-1 mRNA virtually disappeared, suggesting RNAi-like and/or RNase H antisense effects. DMT-1 mRNA levels returned to normal at higher infection levels, indicating that an additional mechanism of mRNA occupation, able to block DMT-1 translation and to avoid feedback regulation by iron responsive elements (IRE), also exists. Cell morphology was normal in all cases and no increases in the interferon-related responses, measured by (a) 2'-5' A oligo synthetase (b) IFITM1 and (c) ISGF3gamma mRNA levels, were observed. Studies presented herein indicate that enterocyte targeting with a gene coding for a short antisense against iron transport blocks enterocyte iron uptake, which may have therapeutic value. PMID- 15896336 TI - Inositol 1,4,5-triphosphate-mediated shuttling between intracellular stores and the cytosol contributes to the sustained elevation in cytosolic calcium in FMLP activated human neutrophils. AB - The current study was designed to probe Ca2+ shuttling between intracellular stores and the cytosol as a potential mechanism contributing to the prolongation of elevated Ca2+ transients in N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP)-activated human neutrophils. Cytosolic Ca2+ concentrations and transmembrane fluxes of the cation were measured using spectrofluorimetric and radiometric procedures, respectively, while inositol 1,4,5-triphosphate (IP3) was measured using a radioreceptor assay. The Ca2+-chelating agent, ethylene glycol bis (beta-aminoethyl ether) N,N,N'N'-tetraacetic acid (EGTA; 10mM), was used to exclude store-operated influx of Ca2+ into neutrophils, while the IP3 receptor antagonist, 2-aminoethoxydiphenyl borate (2-APB, 100 microM), added to the cells 10s after FMLP (0.01 and 1 microM), at which time the increases in IP3 and cytosolic Ca2+ were maximal, was used to eliminate both sustained release from stores and influx of Ca2+. Addition of FMLP at 0.01 or 1 microM resulted in equivalent peak increases in cytosolic Ca2+, while the increase in IP3 was greater and the rate of clearance of Ca2+ from the cytosol slower, in cells activated with 1 microM FMLP. Treatment of the cells with either EGTA or 2-APB following addition of 1 microM FMLP, completely (EGTA) or almost completely (2 APB) abolished the influx of Ca2+ and accelerated the rate of clearance of the cation from the cytosol. Post-peak cytosolic Ca2+ concentrations were lower, and the Ca2+ content of the stores higher, in cells treated with 2-APB. The involvement of IP3 was confirmed by similar findings in cells treated with U 73122 (1 microM), a selective inhibitor of phospholipase C. Taken together, these observations are compatible with IP3-mediated Ca2+ shuttling in neutrophils activated with FMLP. PMID- 15896337 TI - Down-regulatory effect of quercitrin gallate on nuclear factor-kappa B-dependent inducible nitric oxide synthase expression in lipopolysaccharide-stimulated macrophages RAW 264.7. AB - Quercetin 3-O-beta-(2''-galloyl)-rhamnopyranoside (QGR) is a naturally occurring quercitrin gallate, a polyphenolic compound isolated from Persicaria lapathifolia (Polygonaceae). In the present study, QGR compound was discovered to have inhibitory effect on nitric oxide (NO) production in lipopolysaccharide (LPS) stimulated macrophages RAW 264.7. QGR compound attenuated LPS-induced synthesis of both mRNA and protein of inducible nitric oxide synthase (iNOS), in parallel, and inhibited LPS-induced luciferase expression as a reporter of iNOS promoter activity in the macrophages. As a mechanism of the anti-inflammatory action shown by QGR compound, suppression of nuclear factor (NF)-kappaB activation has been documented. QGR compound exhibited inhibitory effect on LPS-mediated NF-kappaB transcriptional activity in macrophages RAW 264.7. Furthermore, the compound inhibited LPS-mediated nuclear translocation of NF-kappaB p65 and DNA binding activity of NF-kappaB complex, in parallel, but did not influence LPS-mediated IkappaBalpha degradation. Taken together, QGR compound suppressed LPS-mediated NF kappaB activation, specifically to nuclear localization step of NF-kappaB p65, which was attributable to its down-regulatory action on LPS-induced NO production and iNOS expression. PMID- 15896338 TI - Relative importance of apoptosis and cell cycle blockage in the synergistic effect of combined R115777 and imatinib treatment in BCR/ABL-positive cell lines. AB - The combination of imatinib and a farnesyltransferase inhibitor might be effective for reducing the number of BCR/ABL-positive leukemia cells. In this study, we examined the differences in the mechanisms of the growth inhibitory effect of the combination of imatinib and R115777 (Zarnestra) among BCR/ABL positive cell lines. Steel and Peckham isobologram analysis indicated that this combination had a strong synergistic inhibitory effect on growth in all imatinib resistant cell lines and their parental cell lines. Levels of cleaved caspase 3 were increased by the combination treatment in all cell lines. However, both the level of cleaved PARP and the number of annexin-V-positive cells were much less increased in KCL22 and KCL22/SR cells than in K562, KU812, K562/SR and KU812/SR cells. The combination treatment promoted p27(KIP1) accumulation and induced a significant increase in the percentage of G0/G1 KCL22 and KCL22/SR cells. In other cell lines, the percentage of G0/G1 cells was not increased but rather decreased. The results indicate that induction of apoptosis and blockage of the cell cycle were major mechanisms of the synergistic inhibitory effect of the combination treatment, but the relative importance of these mechanisms differed among cell types. Additional treatment for overriding the G1 checkpoint may be required to eradicate leukemia cells, in which the combination induces cell cycle arrest. PMID- 15896339 TI - Regulation of CYP26A1 expression by selective RAR and RXR agonists in human NB4 promyelocytic leukemia cells. AB - All-trans retinoic acid (ATRA) can induce complete remission in acute promyelocytic leukemia (APL), but resistance to this treatment develops rapidly partly due to increased ATRA metabolism. Among the cytochrome P450s (CYPs) involved in ATRA metabolism, the ATRA-inducible cytochrome P450 26A1 (CYP26A1) is particularly active although the molecular mechanisms involved in its regulation are not well defined in the target leukemia cells. To study CYP26A1 expression and regulation in APL cells, we used the NB4 promyelocytic leukemia cell line. CYP26A1 constitutive expression was barely detectable in NB4 cells, but ATRA could induce high levels of CYP26A1 expression, which reached a maximum at 72h. To further define CYP26A1 induction mechanisms in the NB4 leukemia cells, we used RARs and RXR selective agonists. The RARalpha agonist BMS753 could elicit maturation, as expected, but not CYP26A1 expression. Treatment with the RARbeta agonist BMS641, or the RARbeta/gamma agonist BMS961, could not elicit maturation, as expected, nor induce CYP26A1 expression. Because CYP26A1 expression could not be induced by RAR ligands alone, NB4 cells were then co-treated with the RXR agonist BMS649. The RXR agonist alone could not induce CYP26A1 expression, nor in combination with either the RARbeta agonist or the RARbeta/gamma agonist. However, the combination of the RXR agonist and the RARalpha agonist could elicit a marked induction of CYP26A1 expression. In conclusion, we have shown that CYP26A1 induction is not essential for the granulocytic maturation of NB4 leukemia cells, and that CYP26A1 induction requires the activation of both RARalpha and RXR in these cells. PMID- 15896340 TI - Morin inhibits 12-O-tetradecanoylphorbol-13-acetate-induced hepatocellular transformation via activator protein 1 signaling pathway and cell cycle progression. AB - Flavonoids are constituents of fruits, vegetables, and plant-derived beverages, as well as components in herbal containing dietary supplements. They exhibit a remarkable spectrum of biochemical and pharmacological activities. In this study, we examined morin (3,5,7,2',4'-pentahydroxyflavone) for its effect on 12-O tetradecanoylphorbol-13-acetate (TPA)-treated human hepatocytes. Morin inhibited TPA-induced cellular transformation in Chang liver cells in a dose-dependent manner. Luciferase assay and electrophoretic mobility shift assay revealed that morin suppressed TPA-induced AP-1 activity, and the inhibition of AP-1 activity by morin was mediated through the inhibition of p38 kinase. Moreover, morin induced the S-phase arrest and inhibited the DNA synthesis in TPA-treated hepatocytes, suggesting that a cell cycle checkpoint was activated by morin to block DNA synthesis in S phase. In conclusion, our results suggested that morin was a potent anti-hepatocellular transformation agent that inhibited cellular transformation by suppressing the AP-1 activity and inducing the S-phase arrest in human hepatocytes. PMID- 15896341 TI - Dicoumarol relieves serum withdrawal-induced G0/1 blockade in HL-60 cells through a superoxide-dependent mechanism. AB - This work was set to study how dicoumarol affects the cell cycle in human myeloid leukemia HL-60 cells. Cells were accumulated in G0/1 after serum deprivation. However, when cells were treated with 5 microM dicoumarol in serum-free medium, a significant increment in the number of cells in S-phase was observed. Inhibition of G0/1 blockade was confirmed by the increase of thymidine incorporation, the phosphorylation of retinoblastoma protein, and the promotion of cell growth in long-term treatments in the absence of serum. Dicoumarol treatment increased superoxide levels, but did not affect peroxide. Increase of cellular superoxide was essential for inhibition of G0/1 blockade, since scavenging this reactive species with a cell-permeable form of SOD and the SOD mimetics 2-amino-3,5 dibromo-N-[trans-4-hydroxycyclohexyl]benzylamine (ambroxol, 100 microM) and copper[II]diisopropyl salicylate (CuDIPS, 10 microM) completely abolished the effect of dicoumarol. However, N-acetyl-cysteine, overexpression of Bcl-2 or a cell-permeable form of catalase were not effective. 5-Methoxy-1,2-dimethyl-3-[(4 nitrophenol)methyl]-indole-4,7-dione (ES936), a mechanism-based irreversible inhibitor of NAD(P)H:quinone oxidoreductase 1 (NQO1), did not promote S phase entry, and dicoumarol still inhibited G0/1 blockade in the presence of ES936. We demonstrate that dicoumarol inhibits the normal blockade in G0/1 in HL-60 cells through a mechanism involving superoxide, but this effect is not dependent solely on the inhibition of the NQO1 catalytic activity. Our results send a precautionary message about use of dicoumarol to elucidate cellular processes involving oxidoreductases. PMID- 15896342 TI - Azaspiracid-4 inhibits Ca2+ entry by stored operated channels in human T lymphocytes. AB - Azaspiracids (AZs) are a new group of phycotoxins discovered in the Ireland coast that includes the isolated analogues: AZ-1, AZ-2, AZ-3, AZ-4 and AZ-5 and the recently described AZ-6-11. Azaspiracid toxic episodes show gastrointestinal illness, but neurotoxic symptoms are also observed in mouse bioassay. Despite their great importance in human health, so far its mechanism of action is largely unknown. In this report, we present the first data about the effect of AZ-4 on cytosolic calcium concentration [Ca2+]i in freshly human lymphocytes. Cytosolic Ca2+ variations were determined by fluorescence digital imaging microscopy using Fura2 acetoxymethyl ester (Fura2-AM). AZ-4 did not modify cytosolic Ca2+ in resting cells. However, the toxin dose-dependent inhibited the increase in cytosolic Ca2+ levels induced by thapsigargin (Tg). AZ-4 decreased Ca2+-influx induced by Tg but did not affect the Ca2+-release from internal stores induced by this drug. The effects of AZ-4 on Ca2+-influx induced by Tg were reversible and not regulated by adenosine 3',5'-cyclic monophosphate (cAMP) pathway. When AZ-4 was added before, after or together with nickel, an unspecific blocker of Ca2+ channels, the effects were indistinguishable and additive. AZ-4 also inhibited maitotoxin (MTX)-stimulated Ca2+-influx by 5-10%. Thus, AZ-4 appeared to be a novel inhibitor of plasma membrane Ca2+ channels, affecting at least to store operated channels, showing an effect clearly different from other azaspiracid analogues. PMID- 15896343 TI - Muscarinic subtype affinity and functional activity profile of 1-methyl-2-(2 methyl-1,3-dioxolan-4-yl)pyrrolidine and 1-methyl-2-(2-methyl-1,3-oxathiolan-5 yl)pyrrolidine derivatives. AB - Starting from two previously studied muscarinic full agonists, characterized by a 1,3-dioxolane ((+)-1) and a 1,3-oxathiolane ((+)-2) cycle, two new series of muscarinic ligands were designed, obtained by the steric complication of the parent compounds produced by freezing the aminoalkyl chain into a pyrrolidine ring. Both tertiary amines and the corresponding iodomethyl derivatives were synthesised and studied, and several compounds of the series which behaved as muscarinic agonists have been selected, on the basis of preliminary binding experiments on rat cortex homogenates, for the present work. Results are presented obtained from testing the affinity of the selected compounds against cloned human muscarinic receptors expressed in CHO cells, in order to evaluate subtype selectivity. Their functional activity on classical models of M1-M4 receptors, in guinea pig and rabbit tissues is also reported. With respect to parent compounds, the new molecules present some selectivity toward hm2 receptors; fair M2 selectivity is also evident in functional studies, where these compounds behave as partial agonists. Among the other compounds of the series (2S, 4'R, 2'S)-1,1-dimethyl-2-(2-methyl-1,3-dioxolan-4-yl)pyrrolidinium iodide ( )-3 and (2R, 5'S, 2'S)-1-methyl-2-(2-methyl-1,3-oxathiolan-5-yl)pyrrolidine (+)-5 present a promising pharmacological profile. Compound (-)-3 shows modest hm2 selectivity in binding experiments but a clearcut M2 selectivity in functional tests, where it behaves as a weak antagonist on M1 and M4 subtypes, as a weak full agonist on the M3 subtype and as a potent partial agonist on M2 subtype. Tertiary amine (+)-5 presents a quite similar profile but appears more interesting since, lacking a permanent charge on the nitrogen atom, it may represent an interesting tool to study CNS muscarinic receptors. Our results confirm that sterical complication of parent compounds (+)-1 and (+)-2 produces more selective muscarinic agonists. PMID- 15896344 TI - Hypoxia induces changes in expression of isoforms of the divalent metal transporter (DMT1) in rat pheochromocytoma (PC12) cells. AB - Although hypoxia has been shown to increase the expression of a variety of proteins involved in iron homeostasis, including transferrin and its receptor, little is known about the effect of low oxygen on formation of isoforms of the major iron transport protein, divalent metal transporter 1, DMT1. Accordingly, we examined the effects of hypoxia on expression and subcellular distribution of the different isoforms of DMT1 in rat PC12 cells. Treatment with low oxygen modestly increased expression of protein and mRNA levels for both the +IRE and -IRE species of DMT1. In contrast, expression of the exon 1A containing species of DMT1 was greatly increased by hypoxia as indicated by Western blot and real-time RT-PCR analysis. Message levels for the 1A isoforms increased approximately 60 fold after exposure of PC12 cells to 1% oxygen for 5 h. The subcellular distribution of exon 1A isoforms of DMT1 remained consistently in the cytoplasmic milieu of the cell after hypoxic exposure, as also did the distribution of +IRE species of DMT1. The -IRE species of DMT1, however, responded to hypoxia by becoming increasingly associated with the regions adjoining the outer cellular membranes, while a portion partially colocalized with an early endosomal marker (EEA). Hypoxia also caused a significant increase in the uptake of manganese in PC12 cells. In summary, these results demonstrate that hypoxia selectively increases expression of exon 1A containing species of DMT1 with lesser increases in either the +IRE or -IRE isoforms the transporter. PMID- 15896345 TI - Reversal of in vitro cellular MRP1 and MRP2 mediated vincristine resistance by the flavonoid myricetin. AB - In the present study, the effects of myricetin on either MRP1 or MRP2 mediated vincristine resistance in transfected MDCKII cells were examined. The results obtained show that myricetin can inhibit both MRP1 and MRP2 mediated vincristine efflux in a concentration dependent manner. The IC50 values for cellular vincristine transport inhibition by myricetin were 30.5+/-1.7 microM for MRP1 and 24.6+/-1.3 microM for MRP2 containing MDCKII cells. Cell proliferation analysis showed that the MDCKII control cells are very sensitive towards vincristine toxicity with an IC50 value of 1.1+/-0.1 microM. The MDCKII MRP1 and MRP2 cells are less sensitive towards vincristine toxicity with IC50 values of 33.1+/-1.9 and 22.2+/-1.4 microM, respectively. In both the MRP1 and MRP2 cells, exposure to 25 microM myricetin enhances the sensitivity of the cells towards vincristine toxicity to IC50 values of 7.6+/-0.5 and 5.8+/-0.5 microM, respectively. The increase of sensitivity represents a reversal of the resistance towards vincristine as a result of MRP1 and MRP2 inhibition. Thus, the present study demonstrates the ability of the flavonoid myricetin to modulate MRP1 and MRP2 mediated resistance to the anticancer drug vincristine in transfected cells, indicating that flavonoids might be a valuable adjunct to chemotherapy to block MRP mediated resistance. PMID- 15896346 TI - Prevention of hemorrhagic shock-induced lung injury by heme arginate treatment in rats. AB - Hemorrhagic shock followed by resuscitation (HSR) induces oxidative stress, which leads to acute lung injury. Heme oxygenase (HO)-1 (EC 1.14.99.3), the rate limiting enzyme in heme catabolism, is inducible by oxidative stress and is thought to play an important role in the protection from oxidative tissue injuries. In this study, we examined expression of HO-1 as well as tissue injuries in the lung, liver, and kidney after HSR in rats. We also pretreated animals with heme arginate (HA), a strong inducer of HO-1, and examined its effect on the HSR-induced lung injury. HO-1 expression significantly increased in the liver and kidney following HSR, while its expression in the lung was very low and unchanged after HSR. In contrast to HO-1 expression, tissue injury and tumor necrosis factor-alpha (TNF-alpha) gene expression was more prominent in the lung compared with those in the liver and kidney. HA pretreatment markedly induced HO 1 in pulmonary epithelial cells, and ameliorated the lung injury induced by HSR as judged by the improvement of histological changes, while it decreased TNF alpha and inducible nitric oxide synthase gene expression, lung wet weight to dry weight ratio, and myeloperoxidase activity. In contrast, inhibition of HO-1 by tin-mesoporphyrin administration abolished the beneficial effect of HA pretreatment. These findings suggest that tissues with higher HO-1 may be better protected than those with lower HO-1 from oxidative tissue injury induced by HSR. Our findings also indicate that HA pretreatment can significantly suppress the HSR-induced lung injury by virtue of its ability to induce HO-1. PMID- 15896347 TI - Cell line dependent RNA expression profiles of prion-infected mouse neuronal cells. AB - The overall impact of prion disease on gene expression is not well characterized. We have carried out a large-scale expression analysis of specific cell types commonly employed in studies of prion disease. Neuroblastoma cells (N2a) and hypothalamic neuronal cells (GT1) can be persistently infected with mouse-adapted scrapie prions, the latter demonstrating cytopathologic effects associated with prion neuropathology. Exploiting a mouse DNA microarray containing approximately 21,000 spotted cDNAs, we have identified several hundred differentially expressed sequences in the two cell lines when infected with prion strain RML. ScN2a and ScGT1 cells demonstrate unique changes in RNA profiles and both differ from the reported changes in human microglia and prion-infected brain studies albeit with some overlap. In addition, several of the identified changes are shared in common with other neurodegenerative diseases such as Alzheimer's disease. The results illustrate that prion infection differs in effect depending on cell type, which could be exploited for diagnostic or therapeutic intervention. PMID- 15896348 TI - Crystal structure of a secreted insect ferritin reveals a symmetrical arrangement of heavy and light chains. AB - Ferritins are iron storage proteins made of 24 subunits forming a hollow spherical shell. Vertebrate ferritins contain varying ratios of heavy (H) and light (L) chains; however, known ferritin structures include only one type of chain and have octahedral symmetry. Here, we report the 1.9A structure of a secreted insect ferritin from Trichoplusia ni, which reveals equal numbers of H and L chains arranged with tetrahedral symmetry. The H/L-chain interface includes complementary features responsible for ordered assembly of the subunits. The H chain contains a ferroxidase active site resembling that of vertebrate H chains with an endogenous, bound iron atom. The L chain lacks the residues that form a putative iron core nucleation site in vertebrate L chains. Instead, a possible nucleation site is observed at the L chain 3-fold pore. The structure also reveals inter- and intrasubunit disulfide bonds, mostly in the extended N terminal regions unique to insect ferritins. The symmetrical arrangement of H and L chains and the disulfide crosslinks reflect adaptations of insect ferritin to its role as a secreted protein. PMID- 15896349 TI - Unfolding a linker between helical repeats. AB - In many multi-repeat proteins, linkers between repeats have little secondary structure and place few constraints on folding or unfolding. However, the large family of spectrin-like proteins, including alpha-actinin, spectrin, and dystrophin, share three-helix bundle, spectrin repeats that appear in crystal structures to be linked by long helices. All of these proteins are regularly subjected to mechanical stress. Recent single molecule atomic force microscopy (AFM) experiments demonstrate not only forced unfolding but also simultaneous unfolding of tandem repeats at finite frequency, which suggests that the contiguous helix between spectrin repeats can propagate a cooperative helix-to coil transition. Here, we address what happens atomistically to the linker under stress by steered molecular dynamics simulations of tandem spectrin repeats in explicit water. The results for alpha-actinin repeats reveal rate-dependent pathways, with one pathway showing that the linker between repeats unfolds, which may explain the single-repeat unfolding pathway observed in AFM experiments. A second pathway preserves the structural integrity of the linker, which explains the tandem-repeat unfolding event. Unfolding of the linker begins with a splay distortion of proximal loops away from hydrophobic contacts with the linker. This is followed by linker destabilization and unwinding with increased hydration of the backbone. The end result is an unfolded helix that mechanically decouples tandem repeats. Molecularly detailed insights obtained here aid in understanding the mechanical coupling of domain stability in spectrin family proteins. PMID- 15896350 TI - Protein misfolding and amyloid formation for the peptide GNNQQNY from yeast prion protein Sup35: simulation by reaction path annealing. AB - We study the early steps of amyloid formation of the seven residue peptide GNNQQNY from yeast prion-like protein Sup35 by simulating the random coil to beta sheet and alpha-helix to beta-sheet transition both in the absence and presence of a cross-beta amyloid nucleus. The simulation method at atomic resolution employs a new implementation of a Langevin dynamics "reaction path annealing" algorithm. The results indicate that the presence of amyloid-like cross-beta sheet strands both facilitates the transition into the cross-beta conformation and substantially lowers the free energy barrier for this transition. This model systems allows us to investigate the energetic and kinetic details of this transition, which is consistent with an auto-catalyzed, nucleation-like mechanism for the formation of beta-amyloid. In particular, we find that electrostatic interactions of peptide backbone dipoles contribute significantly to the stability of the beta-amyloid state. Furthermore, we find water exclusion and interactions of polar side-chains to be driving forces of amyloid formation: the cross-beta conformation is stabilized by burial of polar side-chains and inter residue hydrogen bonds in the presence of an amyloid-like "seed". These findings are in support of a "dry, polar zipper model" of amyloid formation. PMID- 15896351 TI - Graduated driver licensing research in 2004 and 2005. AB - This is the second update of research on graduated driver licensing (GDL) and teenage drivers. It briefly summarizes research in progress and research published since the January 2004 update (Hedlund, J. & Compton, R. [2004]. Graduated driver licensing research in 2003 and beyond. Journal of Safety Research 35(1), 5-11). Research has been very active, especially on teenage driver risk factors, GDL program evaluations, the role of parents in managing and training their teenage drivers, and driver education. Results have strengthened the case for GDL, for nighttime and passenger restrictions, and for extended supervised driving practice. PMID- 15896352 TI - Effects of two doses of methylphenidate on simulator driving performance in adults with attention deficit hyperactivity disorder. AB - INTRODUCTION: Numerous studies have documented an increased frequency of vehicular crashes, traffic citations, driving performance deficits, and driving related cognitive impairments in teens and adults with attention deficit hyperactivity disorder. METHOD: The present study evaluated the effects of two single, acute doses of methylphenidate (10 and 20 mg) and a placebo on the driving performance of 53 adults with ADHD (mean age=37 years, range=18-65) using a virtual reality driving simulator, examiner and self-ratings of simulator performance, and a continuous performance test (CPT) to evaluate attention and inhibition. A double-blind, drug-placebo, within-subjects crossover design was used in which all participants were tested at baseline and then experienced all three drug conditions. RESULTS: A significant beneficial effect for the high dose of medication was observed on impulsiveness on CPT, variability of steering in the standard driving course, and driving speed during the obstacle course. A beneficial effect of the low dose of medication also was evident on turn signal use during the standard driving course. An apparent practice effect was noted on some of the simulator measures between the baseline and subsequent testing sessions that may have interacted with and thereby obscured drug effects on those measures. CONCLUSIONS: The results, when placed in the context of prior studies of stimulants on driving performance, continue to recommend their clinical use as one means of reducing the driving risks in ADHD teens and adults. IMPACT ON INDUSTRY: Given the significantly higher risk of adverse driving outcomes associated with ADHD, industry needs to better screen for ADHD among employees who drive as part of employment so as to improve safety and reduce costs. Use of stimulants to treat the adult ADHD driver may reduce safety risks. PMID- 15896353 TI - Arachidonic acid cascade in endothelial pathobiology. AB - Arachidonic acid (AA) and its metabolites (eicosanoids) represent powerful mediators, used by organisms to induce and suppress inflammation as a part of the innate response to disturbances. Several cell types participate in the synthesis and release of AA metabolites, while many cell types represent the targets for eicosanoid action. Endothelial cells (EC), forming a semi-permeable barrier between the interior space of blood vessels and underlying tissues, are of particular importance for the development of inflammation, since endothelium controls such diverse processes as vascular tone, homeostasis, adhesion of platelets and leukocytes to the vascular wall, and permeability of the vascular wall for cells and fluids. Proliferation and migration of endothelial cells contribute significantly to new vessel development (angiogenesis). This review discusses endothelial-specific synthesis and action of arachidonic acid derivatives with a particular focus on the mechanisms of signal transduction and associated intracellular protein targets. PMID- 15896354 TI - Pigment epithelium-derived factor (PEDF) promotes growth of pericytes through autocrine production of platelet-derived growth factor-B. AB - Microvessels are composed of two types of cells, endothelial cells and pericytes. Pericyte loss or dysfunction participates in various types of disorders, including diabetic retinopathy. Recently, decreased levels of pigment epithelium derived factor (PEDF) in the eye have been found to predict progression of diabetic retinopathy. However, the effect of PEDF on pericyte growth remains to be unknown. In this study, we investigated whether or how PEDF could stimulate proliferation of cultured retinal pericytes. PEDF stimulated DNA synthesis in pericytes in a dose-dependent manner. PEDF up-regulated pericyte mRNA levels of platelet-derived growth factor-B (PDGF-B). Down-regulation of PDGF-B gene expression by small interfering RNAs completely inhibited the PEDF-induced DNA synthesis in pericytes. Furthermore, PEDF increased protein kinase C (PKC) activity in pericytes and staurosporine, a potent cell-permeable inhibitor of PKC, completely blocked the PDGF-B gene induction and subsequent increase in DNA synthesis in PEDF-exposed pericytes. These results demonstrate that PEDF promotes the growth of cultured pericytes possibly through autocrine production of PDGF-B via PKC activation. Our present study suggests that PEDF could act as a mitogen or survival factor for pericytes, thereby being involved in the maintenance of retinal microvascular homeostasis. PMID- 15896355 TI - Insulin enhances leukocyte-endothelial cell adhesion in the retinal microcirculation through surface expression of intercellular adhesion molecule-1. AB - The purpose of this study was to evaluate the effects of insulin on leukocyte endothelial cell adhesion in the retinal microcirculation in vitro and in vivo. Human retinal endothelial cells (HRECs) were cultured in medium with or without insulin, and neutrophils allowed to adhere. Adherent neutrophils were quantified by measuring myeloperoxidase activity. Surface expression of endothelial adhesion molecules were studied with the use of an enzyme immunoassay. Insulin at concentrations of 50 and 100 microU/ml significantly increased neutrophil adhesion to HRECs compared with the control cells (P < 0.01, respectively). Surface expression of intercellular adhesion molecule-1 (ICAM-1) significantly increased when HRECs were exposed to 100 microU/ml insulin, as compared with the control cells (P < 0.05). Anti-ICAM-1 antibody significantly inhibited neutrophils adhesion to HRECs (P < 0.0001). Brown-Norway rats received subcutaneous injection of 0.2 U per 100 g body weight insulin three times. Control rats received the same amount of phosphate-buffered saline. Leukocyte entrapment in the retina was evaluated using acridine orange leukocyte fluorography. The number of leukocytes trapped in the retina of insulin-treated rats was significantly elevated compared with that in the control animals (P < 0.0001). These results suggested that insulin enhances leukostasis in retinal microcirculation. Hyperinsulinemia may be a risk factor of retinal microcirculatory disturbances. PMID- 15896356 TI - Muscle arteriolar and venular reactivity in two models of hypertensive rats. AB - This study was designed to test if skeletal muscle fiber composition could influence vascular response in hypertensive rats. Muscle vessels were observed by intravital microscopy in anesthetized rats and changes in diameter were measured after local administration of endothelium-dependent and -independent vasodilators. Vascular reactivity was compared in two models of hypertension deoxicorticosterone acetate and salt load (DOCA-s) hypertensive rats and spontaneously hypertensive rats (SHR). The muscles used were: the fast-twitch glycolytic muscle, extensor digitorum longus (EDL), and the slow-twitch oxidative, soleus muscle. Maximal dilation induced by vasoactive drugs was of similar magnitude in EDL and soleus arterioles. Terminal arteriole reactivity to acetylcholine and adenosine was blunted in EDL (35% and 49% reduction, respectively) and soleus muscles (42% and 34% reduction, respectively) of SHR compared with Wistar Kyoto rats. Reactivity of DOCA-s rats to acetylcholine, adenosine, and sodium nitroprusside was reduced by 38%, 50%, 39% in EDL third- and fourth-order arterioles and by 30%, 38%, 38% in soleus fourth-order arterioles, respectively. These studies show that hypertension probably induced similar vascular changes in both muscles studied. Vascular reactivity is blunted for some vasodilator drugs and is more affected in DOCA-s rats. In addition, a preferential action for bradykinin was observed on upstream arterioles but not on venules. This effect was not observed for adenosine. PMID- 15896357 TI - The impact of hypercapnia on retinal capillary blood flow assessed by scanning laser Doppler flowmetry. AB - AIM: To determine the effect of hypercapnia on retinal capillary blood flow using scanning laser Doppler flowmetry (SLDF). METHODS: One randomly selected eye of each of 10 normal healthy subjects (mean age 25 years, SD 2.3) was studied. Subjects breathed unrestricted air for 15 min before (baseline) and after raising fractional (percent) end-tidal concentration of CO2 (FETCO2) for 15 min by adding low flows of CO2 to air entering a sequential gas delivery circuit attached to a nasal mask. Five good quality baseline SLDF images were acquired both of the optic nerve head (ONH) and of the macula. Subsequently, a minimum of 7 sequential images were acquired during hypercapnia. Five further images were acquired of the ONH, or of the macula, after returning to unlimited air breathing. The respiratory parameters of subjects were continually monitored. RESULTS: The group mean increase in end-tidal CO2 was 14.13% (SD 4.10) relative to baseline. The nasal macula (P = 0.028) and foveal (P = 0.042) areas showed a significant increase in retinal capillary blood flow in response to hypercapnia while no significant change was noted in the ONH or temporal macula areas. Change in blood flow significantly correlated with change of FETCO2 and/or end-tidal PO2 for 3 of the 4 locations. CONCLUSIONS: Hypercapnia provoked a significant increase in retinal capillary blood flow in 2 of 4 retinal locations. Hypercapnia also induced a change in respiratory parameters that significantly correlated with change in retinal capillary blood flow in 3 of the 4 locations. PMID- 15896358 TI - A design principle for vascular beds: the effects of complex blood rheology. AB - We propose a design principle that extends Murray's original optimization principle for vascular architecture to account for complex blood rheology. Minimization of an energy dissipation function enables us to determine how rheology affects the morphology of simple branching networks. The behavior of various physical quantities associated with the networks, such as the wall shear stress and the flow velocity, is also determined. Our results are shown to be qualitatively and quantitatively compatible with independent experimental observations and simulations. PMID- 15896359 TI - Visualisation of morphological changes in living intact human microvessels using confocal microscopy. AB - Conventional techniques to visualise microvascular structure often involve fixed tissue slices that provide two-dimensional images. A previous study using diffusive labelling of fresh, dissected tissue samples with fluorescently-tagged endothelial markers demonstrated the possibility of examining the three dimensional architecture of the microvasculature using confocal microscopy. The present study extends the use of this quick and simple method of diffusive labelling to examine the possibility of repeatedly measuring changes in the morphology of intact microvessel in response to pharmacological stimuli. Initially, three-dimensional surface-rendered images of the same microvessel derived from the placenta and subcutaneous biopsies demonstrated morphological and topological changes in response to temperature and increasing potassium changes of physiological salt solutions, respectively. Furthermore, a dose response study was performed with subcutaneous microvessels using the potent vasodilator, adrenomedullin. Analysis of a series of z-stack, superimposed to form a single maximum brightness image, demonstrated an inverse dose-response relationship, with responses to increasing adrenomedullin concentrations (10(-12) to 10(-8) M). In vessels that had constricted in response to noradrenaline (diameters: 22.4 to 58.0 microm), physiological concentrations of 10(-12) M increased vessel diameter by 108% above baseline conditions. Control treatment using physiological salt solution did not demonstrate any changes. The technique described suggest that diffusive labelling with vascular endothelial markers such as ulex europeaus agglutinin I in live tissue samples may be used in conjunction with confocal microscopy to demonstrate heterogeneous morphological and topological changes in intact segments of the microvasculature. PMID- 15896360 TI - Use of tadalafil in a patient with a secondary Raynaud's phenomenon not responding to sildenafil. AB - The Raynaud's phenomenon often accompanies systemic rheumatic diseases and is also known as a vascular side effect of chemotherapy. Therapy of the Raynaud's phenomenon with nitrates or calcium-channel-blockers is rarely beneficial. In contrast, the PDE-V-inhibitor sildenafil seems to be effective in these patients. For the first time we report on a patient with Raynaud's phenomenon due to chemotherapy not responding to sildenafil but to the new PDE-V-inhibitor tadalafil in an equivalent dosage. Measurement with a laser Doppler revealed an increased blood flow and a reduction of symptoms. Therefore, therapy of Raynaud's phenomenon with the new PDE-V-inhibitor tadalafil seems to be an effective treatment option in patients not responding to sildenafil. PMID- 15896361 TI - Quantification of hepatic microcirculation heterogeneity of perfusion: effects of endothelin-1. AB - Stress alters hepatic perfusion heterogeneously leading to mismatch of oxygen supply and demand. Endothelin-1 (ET-1) has been linked to stress-mediated microcirculation disruption; however, perfusion heterogeneity is difficult to quantify. Therefore, we developed an image analysis based automated methodology for quantification of heterogeneity (Ht) of perfusion from hepatic intravital microscopy images (HIVM). In vivo assessment of the liver microcirculation flow distribution, before and after ET-1 infusion, was performed in male Sprague Dawley rats using HIVM combined with FITC-labeled RBC infusion. Our method aimed to analyze parallel/spatial and series/temporal intralobular Ht of perfusion as well as interlobular Ht of perfusion and was based on automated RBC detection, tracking, and motion analysis. Compared with manual analysis, this method detected 40 times more RBC without increasing false positives (<0.5%). At baseline, parallel/spatial Ht was higher than series/temporal Ht of perfusion which suggested higher hepatic stellate cells asynchrony between sinusoids than within the same sinusoid. Following ET-1 infusion, parallel/spatial Ht transiently increased contrasting with a decrease in series/temporal Ht. In conclusion, we proposed a HIVM analysis methodology that allows assessment of Ht of perfusion within and between hepatic lobules and improved our understanding of ET-1 mediated effects on the hepatic microcirculation. PMID- 15896362 TI - Global phytochemistry: the Turkish frame. PMID- 15896363 TI - Expression, purification and characterization of recombinant (E)-beta-farnesene synthase from Artemisia annua. AB - A cDNA clone (GenBank Accession No. AY835398) encoding a sesquiterpene synthase, (E)-beta-farnesene synthase, has been isolated from Artemisia annua L. It contains a 1746-bp open reading frame coding for 574 amino acids (66.9 kDa) with a calculated pI=5.03. The deduced amino acid sequence is 30-50% identical with sequences of other sesquiterpene synthases from angiosperms. The recombinant enzyme, produced in Escherichia coli, catalyzed the formation of a single product, beta-farnesene, from farnesyl diphosphate. The pH optimum for the recombinant enzyme is around 6.5 and the K(m)- and k(cat)-values for farnesyl diphosphate, is 2.1 microM and 9.5 x 10(-3) s(-1), respectively resulting in the efficiency 4.5 x 10(-3) M(-1)s(-1). The enzyme exhibits substantial activity in the presence of Mg(2+), Mn(2+) or Co(2+) but essentially no activity when Zn(2+), Ni(2+) or Cu(2+) is used as cofactor. The concentration required for maximum activity are estimated to 5 mM, 0.5 mM and <10 microM for Mg(2+), Co(2+) or Mn(2+), respectively. Geranyl diphosphate is not a substrate for the recombinant enzyme. PMID- 15896364 TI - Purification and characterization of an allosteric fructose-1,6-bisphosphate aldolase from germinating mung beans (Vigna radiata). AB - Cytosolic fructose-1,6-P(2) (FBP) aldolase (ALD(c)) from germinated mung beans has been purified 1078-fold to electrophoretic homogeneity and a final specific activity of 15.1 micromol FBP cleaved/min per mg of protein. SDS-PAGE of the final preparation revealed a single protein-staining band of 40 kDa that cross reacted strongly with rabbit anti-(carrot ALD(c))-IgG. The enzyme's native M(r) was determined by gel filtration chromatography to be 160 kDa, indicating a homotetrameric quaternary structure. This ALD is a class I ALD, since EDTA or Mg(2+) had no effect on its activity, and was relatively heat-stable losing 0-25% of its activity when incubated for 5 min at 55-65 degrees C. It demonstrated: (i) a temperature coefficient (Q(10)) of 1.7; (ii) an activation energy of 9.2 kcal/mol active site; and (iii) a broad pH-activity optima of 7.5. Mung bean ALD(c) is bifunctional for FBP and sedoheptulose-1,7-P(2) (K(m) approximately 17 microM for both substrates). ATP, ADP, AMP and ribose-5-P exerted inhibitory effects on the activity of the purified enzyme. Ribose-5-P, ADP and AMP functioned as competitive inhibitors (K(i) values=2.2, 3.1 and 7.5mM, respectively). By contrast, the addition of 2mM ATP: (i) reduced V(max) by about 2-fold, (ii) increased K(m)(FBP) by about 4-fold, and (iii) shifted the FBP saturation kinetic plot from hyperbolic to sigmoidal (h=1.0 and 2.6 in the absence and presence of 2mM ATP, respectively). Potent feedback inhibition of ALD(c) by ATP is suggested to help balance cellular ATP demands with the control of cytosolic glycolysis and respiration in germinating mung beans. PMID- 15896365 TI - Potato tuber isoapyrases: substrate specificity, affinity labeling, and proteolytic susceptibility. AB - Apyrase/ATP-diphosphohydrolase hydrolyzes di- and triphosphorylated nucleosides in the presence of a bivalent ion with sequential release of orthophosphate. We performed studies of substrate specificity on homogeneous isoapyrases from two potato tuber clonal varieties: Desiree (low ATPase/ADPase ratio) and Pimpernel (high ATPase/ADPase ratio) by measuring the kinetic parameters K(m) and k(cat) on deoxyribonucleotides and fluorescent analogues of ATP and ADP. Both isoapyrases showed a broad specificity towards dATP, dGTP, dTTP, dCTP, thio-dATP, fluorescent nucleotides (MANT-; TNP-; ethene-derivatives of ATP and ADP). The hydrolytic activity on the triphosphorylated compounds was always higher for the Pimpernel apyrase. Modifications either on the base or the ribose moieties did not increase K(m) values, suggesting that the introduction of large groups (MANT- and TNP-) in the ribose does not produce steric hindrance on substrate binding. However, the presence of these bulky groups caused, in general, a reduction in k(cat), indicating an important effect on the catalytic step. Substantial differences were observed between potato apyrases and enzymes from various animal tissues, concerning affinity labeling with azido-nucleotides and FSBA (5'-p fluorosulfonylbenzoyl adenosine). PLP-nucleotide derivatives were unable to produce inactivation of potato apyrase. The lack of sensitivity of both potato enzymes towards these nucleotide analogues rules out the proximity or adequate orientation of sulfhydryl, hydroxyl or amino-groups to the modifying groups. Both apyrases were different in the proteolytic susceptibility towards trypsin, chymotrypsin and Glu-C. PMID- 15896366 TI - Grifolisin, a member of the sedolisin family produced by the fungus Grifola frondosa. AB - The pepstatin-insensitive carboxyl proteinase grifolisin was purified from fruiting bodies of the fungus Grifola frondosa, a maitake mushroom. The enzyme had an optimum pH of 3.0 for the digestion of hemoglobin and 2.8 for milk casein digestion. Its molecular mass was determined to be 43kDa by SDS-PAGE and 40kDa by gel chromatography on Superose 12, and its isoelectric point was found to be 4.6 by isoelectric focusing. The enzyme hydrolyzed four major bonds in the oxidized insulin B-chain: Phe1-Val2, Ala14-Leu15, Gly20-Glu21 and Phe24-Phe25 at pH 3.0. The first 15 amino acid residues in the N-terminal region were AVPSSCASTITPACL, and the coding region of the grifolisin gene (gfrF) has a 1960-base pair cDNA. The predicted mature grifolisin protein consisted of 365 residues and was 26% identical to that of sedolisin from Pseudomonas sp. 101 and 34% identical to that of aorsin from Aspergillus oryzae. Grifolisin is a member of the sedolisin S53 family and is not inhibited by pepstatin. PMID- 15896367 TI - A comparison of two strategies to modify the hydroxylation of condensed tannin polymers in Lotus corniculatus L. AB - A full-length sense Antirrhinum majus dihydroflavonol reductase (DFR) sequence was introduced into birdsfoot trefoil (Lotus corniculatus L.) in experiments aimed at modifying condensed tannin content and polymer hydroxylation in a predictable manner. Analysis of transgenic plants indicated lines that showed enhanced tannin content in leaf and stem tissues. In contrast to previous data from root cultures, levels of propelargonidin units were not markedly elevated in lines with enhanced tannin content. RT-PCR analysis of four selected lines indicated a correlation between enhanced tannin content and expression of the introduced DFR transgene. Using a contrasting approach we introduced a flavonoid 3'5' hydroxylase (F3'5'H) sequence derived from Eustoma grandiflorum into Lotus root cultures. Expression of the transgene was associated with increased levels of condensed tannins and in this case there was also no alteration in polymer hydroxylation. These results suggest that additional mechanisms may exist that control the hydroxylation state of condensed tannins in this model species. PMID- 15896368 TI - Brassinosteroids are inherently biosynthesized in the primary roots of maize, Zea mays L. AB - GC-MS analysis revealed that primary roots of maize contain 6-deoxocathasterone, 6-deoxoteasterone and 6-deoxotyphasterol. These brassinosteroids, and the previously identified campesterol, campestanol, 6-deoxocastasterone and castasterone, in the roots are members of a biosynthetic pathway to castasterone, namely the late C-6 oxidation pathway, suggesting that its biosynthetic pathway is operative in the roots. To verify this, a cell-free enzyme extract was prepared from maize roots, and enzymatic conversions from campesterol to castasterone through the aforementioned sterols and brassinosteroids were examined. The presence for the biosynthetic sequences, campesterol-->24 methylcholest-4-en-3beta-ol-->24-methylcholest-4-en-3-one-->24-methylcholest-5 alpha-cholestan-3-one-->campestanol and 6-deoxoteasterone-->6-deoxo-3 dehydroteasterone-->6-deoxotyphasterol-->6-deoxocastasterone-->castasterone were demonstrated. These results indicate that maize roots contain a complete set of enzymes involved in the late C-6 oxidation pathway, thereby demonstrating that endogenous brassinosteroids are biosynthesized in the roots. PMID- 15896369 TI - Metabolism of daidzein by Nocardia species NRRL 5646 and Mortierella isabellina ATCC 38063. AB - The phytoestrogen daidzein was metabolized by Nocardia species NRRL 5646 to give two metabolites obtained by hydroxylation and methylation. These metabolites were spectrally characterized as 7-methoxy-4'-hydroxyisoflavone (isoformononetin) and 7,8-dimethoxy-4'-hydroxyisoflavone. Mortierella isabellina ATCC 38063 was able to metabolize daidzein to the unusual metabolite daidzein-4'-rhamnopyranoside. PMID- 15896370 TI - Phytotoxins from the fungus Malbranchea aurantiaca. AB - Bioassay-directed fractionation of an ethyl acetate extract from cultures of the fungus Malbranchea aurantiaca led to the isolation of two phytotoxic compounds, namely, 1-hydroxy-2-oxoeremophil-1(10),7(11),8(9)-trien-12(8)-olide (1) and penicillic acid (2). The structure of 1 was established by spectroscopic and X ray crystallographic analyses. Metabolites 1 and 2 caused significant inhibition of radicle growth of Amaranthus hypochondriacus with IC(50) values 6.57 and 3.86 microM, respectively. In addition, 1 inhibited activation of the calmodulin dependent enzyme cAMP phosphodiesterase (IC(50)=10.2 microM). PMID- 15896371 TI - Inhibitory effects of Piper umbellatum and Piper peltatum extracts towards myotoxic phospholipases A2 from Bothrops snake venoms: isolation of 4 nerolidylcatechol as active principle. AB - Phospholipases A(2) (PLA(2)) are important constituents of snake venoms, being responsible for several of their toxic actions. Extracts from plants used in folk medicine were screened for inhibition of the enzymatic activity of myotoxin I, a PLA(2) from Bothrops asper. Piper umbellatum and Piper peltatum extracts tested positive, and their fractionation resulted in the isolation of 4 nerolidylcatechol. Its inhibitory effects towards toxic activities of two Bothrops myotoxins, representing catalytically active (Asp49) and catalytically inactive (Lys49) types of group II PLA(2)s, respectively, were characterized. The enzyme activity of B. asper myotoxin I was completely inhibited by 4 nerolidylcatechol at an inhibitor:toxin ratio of 10:1 (wt/wt) with an IC50 of approximately 1mM. In addition, 4-nerolidylcatechol inhibited representatives of groups I and III of PLA(2)s. Its preincubation with Bothrops myotoxins significantly reduced their myotoxic and edema-inducing activities in animal experiments. However, when 4-nerolidylcatechol was administered in situ, immediately after toxin injection, its inhibitory ability was substantially lower or negligible. This might be explained by the rapid action of these toxins in vivo, together with the slow inactivation of PLA(2) activity observed in vitro. Electrophoretic and chromatographic analyses of myotoxins ruled out major changes in protein charge, hydrophobicity, or gross molecular mass being involved in the inhibition mechanism. Mass spectrometry determinations are consistent with the covalent modification of myotoxin by one molecule of 4-nerolidylcatechol. Finally, a novel compound was isolated from both Piper species, sharing the nerolidyl skeleton, but nevertheless not being inhibitory towards the PLA(2)s studied. PMID- 15896372 TI - High molecular weight constituents from roots of Echinacea pallida: an arabinogalactan-protein and an arabinan. AB - This investigation shows structural features of two macromolecules from roots of Echinacea pallida (Nutt.) Nutt: an arabinogalactan-protein (AGP) and an arabinan. The arabinogalactan-protein was precipitated with beta-glucosyl Yariv reagent from a high molecular weight fraction. Investigations of the neutral sugar composition revealed Gal (52.1% w/w) and Ara (38.2% w/w) in a ratio of 1.4:1, accompanied by Glc (6.9% w/w) and Rha (2.8% w/w). The content of uronic acids was 6.2%. Mild acid hydrolysis detects Ara and Glc being located at the periphery of the molecule. Linkage analyses and NMR spectroscopy revealed a backbone of the polysaccharide mainly consisting of 3-linked and 3,6-linked Galp-residues. Side chains are composed of 3,6-linked or 6-linked Galp terminating in 5-linked Araf, terminal Araf, Glcp and GlcAp. The protein part (3.9% w/w) of the AGP is rich in Hyp, Ser, Ala, Thr, Glu, Asp and Gly. The amount of Hyp was determined by a colorimetric method and found to be (0.65% (w/w) of the AGP, which is in good agreement with the result obtained by amino acid hydrolysis (0.67% w/w). The arabinan was isolated from the supernatant of the Yariv precipitation on the basis of solubility in EtOH (80%). It mainly consists of Ara (85.8%). Linkage analyses and NMR spectroscopy indicate a highly branched molecule, consisting of 3,5-linked, 5-linked and terminal Araf-residues in equal amounts. PMID- 15896373 TI - Lipophilic flavones of Primula veris L. from field cultivation and in vitro cultures. AB - Ten lipophilic flavones were isolated from the leaves of Primula veris from field cultivation - the newly described 3'-hydroxy-4',5'-dimethoxyflavone and 3' methoxy-4',5'-methylenedioxyflavone, the previously known from chemical synthesis 3',4'-dimethoxyflavone, 2',5'-dimethoxyflavone, and also flavone, 2' hydroxyflavone, 2'-methoxyflavone, 3'-methoxyflavone, 3',4',5'-trimethoxyflavone and 5,6,2',6'-tetramethoxyflavone (zapotin) which were previously known from plants. The same flavones were found in the leaves of P. veris obtained by in vitro propagation. The structural assignments were derived from (1)H NMR, (13)C NMR, EIMS and UV spectral data and the influence of B-ring oxygen substituents on the C-2, C-3 and H-3 NMR resonances in flavones unsubstituted in the A ring is taken into consideration. PMID- 15896374 TI - Marsdenosides A-H, polyoxypregnane glycosides from Marsdenia tenacissima. AB - Eight polyoxypregnane glycosides, marsdenosides A-H, were isolated from the CHCl(3)-soluble fraction of the ethanolic extract of the stem of Marsdenia tenacissima, along with six known glycosides and two known polyoxypregnane aglycones. Three polyoxypregnanes, 12beta-O-2-methylbutyryl-tenacigenin A, 11alpha,12beta-di-O-acetyltenacigenin B, and 11alpha-O-tigloyltenacigenin B were also obtained. Their structures were established on the basis of spectroscopic analysis and chemical evidence. PMID- 15896375 TI - Terrestrins A-G: p-terphenyl derivatives from the inedible mushroom Thelephora terrestris. AB - Seven p-terphenyl derivatives named terrestrins A-G together with three known ganbajunin B, thelephantins F and H, were isolated from the methanol extract of fruiting bodies of the Japanese inedible mushroom Thelephora terrestris (Thelephoraceae). Their structures were elucidated by means of high-resolution MS, 2D NMR, IR and UV spectroscopy, and X-ray crystallographic analysis. PMID- 15896376 TI - Labdane diterpenes from Marrubium velutinum and Marrubium cylleneum. AB - From the aerial parts of Marrubium velutinum and Marrubium cylleneum, seven labdane diterpenes, velutine A, 15-epi-velutine A, velutine B, 15-epi-velutine B, velutine C, cyllenine A and 15-epi-cyllenine A, have been isolated together with five known diterpenes and four known flavones. The structures of the isolated compounds were established by means of NMR [(1)H-(1)H-COSY, (1)H-(13)C-HMQC, HMBC, HMQC-TOCSY, NOESY] and MS spectral analyses. Complete NMR assignments are reported for known compounds. PMID- 15896377 TI - Characterization of cell wall polysaccharides from the medicinal plant Panax notoginseng. AB - Panax notoginseng is a commonly used medicinal plant in south-western China. Recent studies indicate that wall polysaccharides are responsible for some of the immunostimulatory activity. Fractionation of the P. notoginseng root powder alcohol insoluble residue (AIR) and its compositional analysis enabled us to deduce the polysaccharide and protein composition of the root cell walls. P. notoginseng walls are composed primarily of polysaccharide (approximately 97% w/w) and some protein. The polysaccharides include pectic polysaccharides (neutral Type I 4-galactan (21%), arabinan (5%), acidic rhamnogalacturonan I (RG I, 2%) and homogalacturonan (HGA, 24%), non-cellulosic polysaccharides (heteroxylan, 3%), xyloglucan (XG, 3%) and heteromannan (1%)) and cellulose (24%). The root AIR also contains Type II AG/AGPs (5% w/w) typically associated with the plasma membrane and extracellular matrix. Thus, P. notoginseng roots contain polysaccharides typical of Type I primary cell walls but are distinguished by their very high levels of Type I 4-galactans and low levels of XGs. The major amino acids in the AIR were Leu (14 mol%), Asx (16 mol%), Glx (10 mol%), Ala (9 mol%), Thr (9 mol%) and Val (9 mol%). PMID- 15896378 TI - Cyclolignans from Scyphocephalium ochocoa via high-throughput natural product chemistry methods. AB - Two 2,7'-cyclolignans, ocholignans A and B, were obtained as mass-limited samples from Scyphocephalium ochocoa via high-throughput natural products chemistry methods. The rapid structure elucidation of each compound was primarily facilitated by NMR data acquisition using a capillary-scale NMR probe, CapNMR probe. Ocholignan A was found to possess significant in vitro antibacterial activity against Gram-positive bacteria methicillin-resistant Staphylococcus aureus ATCC 33591 and S. aureus 78-13607A with a MIC of 16 microg/mL, respectively. PMID- 15896379 TI - The crucial role of tense for verb production. AB - The case of an aphasic patient whose spontaneous speech contains very few lexical verbs is reported. Instead of sentences with lexical verbs, the patient produces many (grammatical) copular constructions. He also substitutes lexical verbs with the copula. Although this results in ungrammatical utterances, by doing so, a resemblance of sentence structure and a degree of grammaticality of his utterances are preserved. Although the patient is more impaired in naming action than object pictures, it is unlikely that lexical retrieval difficulties are solely responsible for the paucity of lexical verbs in his speech. A series of tests revealed a profound deficit in producing tense marking inflections and in understanding their significance. We argue that the unavailability of tense features is the primary reason for the lack of lexical verbs in his speech. An alternative possibility, that the tense deficit interacts with the verb retrieval deficit, is also discussed. The patient has a complex lesion and language profile, with features associated with both Broca's and Wernicke's aphasia. However, since the study focuses on his verb and tense deficits and the grammaticality of his utterances, issues that are often discussed in relation to agrammatic Broca's aphasia, the literature that is relevant to these topics and to Broca's aphasia is reviewed, despite the different diagnostic profile of the patient. PMID- 15896381 TI - Detection of sound rise time by adults with dyslexia. AB - Low sensitivity to amplitude modulated (AM) sounds is reported to be associated with dyslexia. An important aspect of amplitude modulation cycles are the rise and fall times within the sound. In this study, simplified stimuli equivalent to just one cycle were used and sensitivity to varying rise times was explored. Adult participants with dyslexia or compensated dyslexia and a control group performed a detection task with sound pairs of different rise times. Results showed that the participants with dyslexia differed from the control group in rise time detection and a correlation was found between rise time detection and reading and phonological skills. A subgroup of participants with lower sensitivity to rise time detection characterized by low accuracy in syllable level phonological skills was found within the dyslexic group. Short stimuli containing only one rise time produced associations with phonological skills and reading, even in a language where the perception of rise time contrasts are not crucial for the signaling of phonemic contrast. PMID- 15896380 TI - The Boston Naming Test in Swedish: normative data. AB - The purpose of the present study was to introduce a Swedish version of the Boston Naming Test and to offer normative data based on a sample of native Swedish speaking healthy adults stratified concerning age, gender, and length of education. The subjects were assessed with other lexical tests and half of the group also performed tests of global cognitive function. A semantic analysis of the responses was performed and the typical Swedish naming of the pictures of BNT was investigated. The results showed that long education, high performance on tests of global cognitive ability (FSIQ), verbal fluency, and other lexical tests had a significant positive association to a good performance on BNT whereas age and gender had a minor influence. The response analysis showed that the more difficult an item was, the more variations in the response pattern. To conclude, the Boston Naming Test is appropriate for use in a Swedish-speaking context and an effective test for assessment of naming ability. PMID- 15896382 TI - Patterns of comprehension performance in agrammatic Broca's aphasia: a test of the Trace Deletion Hypothesis. AB - We tested the core prediction of the Trace Deletion Hypothesis (TDH) of agrammatic Broca's aphasia, which contends that such patients' comprehension performance is normal for active reversible sentences but at chance level for passive reversible sentences. We analyzed the comprehension performance of 38 Italian Broca's aphasics with verified damage to Broca's area, who completed sentence-to-picture matching tasks using active and passive reversible sentences as stimuli. The results failed to confirm the predictions made by TDH: only a small minority (15%) performed at chance on passive sentences and better than chance on active sentences. Furthermore, the distribution of the 38 subjects' performance on passive sentences differed from that predicted by the TDH since many more subjects performed at better-than-chance levels than expected. We discuss the implication of these results for claims about the distribution of language processing mechanisms in the brain. PMID- 15896383 TI - Fluent versus nonfluent primary progressive aphasia: a comparison of clinical and functional neuroimaging features. AB - To better characterize fluent and nonfluent variants of primary progressive aphasia (PPA). Although investigators have recognized both fluent and nonfluent patients with PPA, the clinical and neuroimaging features of these variants have not been fully defined. We present clinical and neuropsychological data on 47 PPA patients comparing the fluent (n=21) and nonfluent (n=26) subjects. We further compared language features with PET/SPECT data available on 39 of these patients. Compared to the nonfluent PPA patients, those with fluent PPA had greater impairment of confrontational naming and loss of single word comprehension. They also exhibited semantic paraphasic errors and loss of single word comprehension. Patients with nonfluent PPA were more likely to be female, were more often dysarthric, and exhibited phonological speech errors in the absence of semantic errors. No significant differences were seen with regard to left hemisphere abnormalities, suggesting that both variants result from mechanisms that overlap frontal, temporal, and parietal regions. Of the language measures, only semantic paraphasias were strongly localized, in this case to the left temporal lobe. Fluent and nonfluent forms of PPA are clinically distinguishable by letter fluency, single word comprehension, object naming, and types of paraphasic errors. Nevertheless, there is a large amount of overlap between dysfunctional anatomic regions associated with these syndromes. PMID- 15896384 TI - Category-specific naming effect in non-brain-damaged individuals. AB - A category effect (i.e., living vs. nonliving exemplars) in confrontation naming has been reported in association with various cerebral pathologies. However, the published reports conflict as to the presence of this category effect in normal controls. The present experiment included 90 subjects in three age groups (young, young-elderly, and old-elderly) and controlled the stimuli in two major ways: by increasing the difficulty level to avoid a ceiling effect, and by equating the stimulus lists on five important word property variables (word frequency, familiarity, age of acquisition, visual complexity, and name agreement). The results confirmed that the category effect was present in the two elderly groups but not in the younger group. However, a gender effect was evidenced in the younger group but not in the two elderly groups. Overall, these results suggest that the category effect reported in neurological populations represents at least in part the influence of normal aging on semantic memory. PMID- 15896385 TI - Discrimination training of phonemic contrasts enhances phonological processing in mainstream school children. AB - Auditory perceptual learning has been proposed as effective for remediating impaired language and for enhancing normal language development. We examined the effect of phonemic contrast discrimination training on the discrimination of whole words and on phonological awareness in 8- to 10-year-old mainstream school children. Eleven phonemic contrast continua were synthesised using linear interpolation coding from real speaker endpoints. Thirty children were pre-tested on the Word Discrimination Test (WDT) and the Phonological Assessment Battery (PhAB). Eighteen then trained for 12 x 30min sessions over 4 weeks using an adaptive three interval two alternative phonemic matching task. The remaining children participated in regular classroom activities. In Post-testing, trained children significantly increased their age-equivalent scores on both the WDT and PhAB by about 2 years. For the PhAB, no improvement was found in the controls. Enhanced performance in the trained children was maintained in a delayed test 5-6 weeks following training. Enhancements on the trained discriminations were weak and variable. The results indicate a dramatic improvement in phonological awareness following phonemic discrimination training without matching perceptual learning. PMID- 15896386 TI - Semantic processing of living and nonliving concepts across the cerebral hemispheres. AB - Studies of patients with category-specific semantic deficits suggest that the right and left cerebral hemispheres may be differently involved in the processing of living and nonliving domains concepts. In this study, we investigate whether there are hemisphere differences in the semantic processing of these domains in healthy volunteers. Based on the neuropsychological findings, we predicted a disadvantage for nonliving compared to living concepts in the right hemisphere. Our prediction was supported, in that semantic decisions to nonliving concepts were significantly slower and more error-prone when presented to the right hemisphere. In contrast there were no hemisphere differences for living concepts. These findings are consistent with either differential representation or processing of concepts across right and left hemispheres. However, we also found a disadvantage for nonliving things compared to living things in the left hemisphere, which is not consistent with a simple representation account. We discuss these findings in terms of qualitatively different semantic processing in right and left hemispheres within the framework of a distributed model of conceptual representation. PMID- 15896387 TI - On the facilitatory effects of cognate words in bilingual speech production. AB - There is a growing body of evidence showing that a word's cognate status is an important dimension affecting the naming performance of bilingual speakers. In a recent article, Kohnert extended this observation to the naming performance of an aphasic bilingual (DJ). DJ named pictures with cognate names more accurately than pictures with non-cognate names. Furthermore, having named the pictures in Spanish helped the subsequent retrieval (with a delay of one week between the two tests) of the same pictures' names in English, but only for pictures with cognate names. That is, there was a language transfer but only for those translation words that were phonologically similar. In this article we first evaluate the conclusions drawn from these results by Kohnert, and second we discuss the theoretical implications of the facilitatory effects of cognate words for models of speech production in bilingual speakers. PMID- 15896388 TI - A structural account of phonological paraphasias. AB - The aim of this study is to develop a partial theory of phonological paraphasias which has some cross-syndrome and cross-linguistic validity. It is based on the distinction between content and structural units and emphasizes the role of the latter. The notion of structure holds the key to an understanding of the differences among the following three types of data: slips of the tongue, slips of the pen (defined as errors committed by healthy, competent adults), and paraphasias. A comparison of these three data types reveals that slips of the pen and paraphasias display striking similarities whereas slips of the tongue stand apart. This pattern emerges very clearly in an analysis of nine empirical effects all of which are found to be of a structural nature. On the basis of these results, it is argued that both the written output of normals and the oral output of aphasics are generated under a reduced structural representation (i.e., weak activation of structural nodes). However, the reasons for the diminished sensitivity to structure are not the same in the two modalities. While the impoverished structural representation is a likely consequence of the relatively slow production rate in writing, it may stem from an impaired transmission of activation among structural nodes in aphasia. By contrast, the normal speaking process occurs under the sway of a full-fledged structural representation. Hence, slips of the tongue are highly sensitive to structural effects. PMID- 15896389 TI - The role of sensory-motor information in object recognition: evidence from category-specific visual agnosia. AB - The role of sensory-motor representations in object recognition was investigated in experiments involving AD, a patient with mild visual agnosia who was impaired in the recognition of visually presented living as compared to non-living entities. AD named visually presented items for which sensory-motor information was available significantly more reliably than items for which such information was not available; this was true when all items were non-living. Naming of objects from their associated sound was normal. These data suggest that both information about object form computed in the ventral visual system as well as sensory-motor information specifying the manner of manipulation contribute to object recognition. PMID- 15896390 TI - Syntactic development in children with hemispherectomy: the I-, D-, and C systems. AB - This study reports on functional morpheme (I, D, and C) production in the spontaneous speech of five pairs of children who have undergone hemispherectomy, matching each pair for etiology and age at symptom onset, surgery, and testing. Our results show that following left hemispherectomy (LH), children evidence a greater error rate in the use of functional category elements than their right hemispherectomy (RH) counterparts. Nevertheless, error rates are surprisingly low and comparable across groups. We interpret these results as (a) weak empirical evidence for a left hemisphere advantage in acquisition of functional structure, (b) strong support that functional structure is a property of all human grammars, and (c) strong support that each isolated developing hemisphere has the potential to acquire a grammar embodying and constrained by highly specific structural principles defining human language. PMID- 15896391 TI - Identifying lesions on structural brain images--validation of the method and application to neuropsychological patients. AB - The study of neuropsychological disorders has been greatly facilitated by the localization of brain lesions on MRI scans. Current popular approaches for the assessment of MRI brain scans mostly depend on the successful segmentation of the brain into grey and white matter. These methods cannot be used effectively with large lesions because lesions usually impair segmentation. We propose a novel, fully automated approach for the delineation of brain lesions on MR scans. This method involves comparing a skull stripped, smoothed, unsegmented T1 images to a control group using the general linear model. We tested this method by using images with simulated lesions of different sizes and images containing real lesions from patients with language deficits. We also tested how varying the size of the Gaussian smoothing kernel affects detection. The simulation was informed by findings of a lesion morphological study also presented here. The proposed method detected simulated lesions effectively in the range of 30--90% 0.05). None of the participants in this study had used a discussion forum before and whilst some students had the skills and confidence to contribute to the on-line discussions, others 'lurked' and some did not access the discussion facility at all. Strategies for improving the engagement and quality of on-line learning are proposed from the lessons learned during this study. PMID- 15896412 TI - Nursing students' and tutors' perceptions of learning and teaching in a clinical skills centre. AB - BACKGROUND: Clinical Skills Centres (CSCs) can ease pressure on clinical skills development and assessment in clinical areas; and provide added value through experiential learning and self-directed learning. Published accounts of innovation in CSCs tell part of this story but little is known about perceptions of students and tutors engaged in day-to-day learning and teaching in CSCs. METHODS: This paper reports one strand of a mixed methods study in a busy multidisciplinary CSC: a questionnaire survey of nursing students' and tutors' perceptions of learning and teaching. Questionnaires focused on items representing commonly espoused views regarding the use and usefulness of CSCs. RESULTS: Students and tutors enjoyed learning and teaching within the CSC, although senior students were slightly muted in their views. All groups valued the supported practice of clinical and communication skills. The CSC was seen as a learning environment that supports the linking of theory and practice. There was some ambivalence, particularly among tutors, about the relationship between performance in the CSC and in clinical areas. DISCUSSION: The favoured pedagogic approach of expert demonstration by tutors followed by supported practice necessitates attention to tutors' training. The muted responses of senior students may signal a need to review the CSC learning experiences offered to them. PMID- 15896413 TI - Improving research supervision in nursing. AB - In this paper, four experienced researchers from the UK, China and Australia offer guidance in research supervision based on their experiences and the recent document, Improving standards in postgraduate research degree programmes [Higher Education Funding Council for England, 2003. Improving standards in postgraduate research degree programmes. Formal consultation. Department for Employment and Learning, Northern Ireland, Higher Education Funding Council for England, Higher Education Funding Council for Wales, Scottish Higher Education Funding Council, HEFCE, London]. Supervision is an important aspect of not only the development of the neophyte researcher, but of academic staff and research activity in general. With increased academic accountability, good supervision should be an integral component of a quality research governance framework and resourced as such. Recommendations include: adoption of these standards; rigorous selection of research students and supervisors and development of projects; development of departmental procedures for monitoring, feedback and intellectual property; and transparency, rigour and fairness in examination procedures. PMID- 15896414 TI - Strategies to overcome obstacles in the facilitation of critical thinking in nursing education. AB - This paper seeks to describe strategies that can be used to overcome obstacles in the facilitation of critical thinking in nursing education. A qualitative, explorative, descriptive and contextual design was used to conduct the research in which fourth year basic comprehensive students and nurse educators volunteered to take part in the study by signing an informed consent. The participants were purposively selected. Focus group interviews were used to collect data from both groups. Tesch's descriptive method of open coding described in (Creswell, J. 1994. RESEARCH DESIGN: Qualitative and quantitative approach. Sage, London.) was used to analyse data. (Democratic Nursing Organisation of South Africa, 1998. Ethical standards for nurse researchers. Denosa, Pretoria) ethical standards for research were observed to maintain the standard and quality of the research. (Lincoln, Y.S., Guba, E.G. 1985. Naturalistic inquiry. Sage, London.) framework was used to ensure trustworthiness of the study. The following obstacles were identified and recontextualised within the existing literature to be able to describe the strategies to overcome the identified obstacles to the facilitation of critical thinking of students: the educators' lack of knowledge; use of teaching and assessment methods that do not facilitate critical thinking of learners; the negative attitudes of educators and their resistance to change; inappropriate selection process and poor educational background that did not facilitate critical thinking of students; inadequate socialisation, cultural and instructional language incompetence. Findings indicated that there is a need for nurse educators to model critical thinking in all aspects of nursing education. It is recommended that there be a whole paradigm shift in nursing education from the traditional teacher-centred methods to a more learner-centred approach that will facilitate critical thinking of student nurses. PMID- 15896415 TI - Student learning in clinical nursing education: perceptions of the relationship between assessment and learning. AB - There is a wealth of literature demonstrating that clinical nursing education is an important part of the baccalaureate programme in preparing students for entry into the nursing profession. While much attention has been given to the factors that can affect learning outcomes in the clinical environment, student and teacher perceptions of the relationship between assessment and learning has remained an under-researched area. The purpose of this study was to explore the perceptions about what students learned and how they learned during their clinical practicum, and to examine the role played by assessment in influencing student learning. Data were collected through a series of focus group interviews with groups of nursing students, graduates, and teachers. It was revealed that students' learning during the clinical practicum was, to a large extent, affected by their perceptions of the assessment tasks. As a result, they adopted a surface approach to learning and focused on preparing for the assessment tasks to the detriment of their learning. Assessment, in this study, exerted what has been described as a negative "backwash" effect on learning. Since assessment may also foster student learning in a positive way, suggestions are offered as to what can be done to bring about a positive "backwash" effect. PMID- 15896416 TI - An exploration of student midwives' language to describe non-formal learning in professional practice. AB - The essence of non-formal learning in midwifery practice has not been previously explored. This paper provides an in-depth analysis of the language of a sample of student midwives' descriptions of their practice learning in a range of clinical settings. The students submitted audio-diaries as part of a national study (Pope, R., Graham. L., Finnerty. G., Magnusson, C. 2003. An investigation of the preparation and assessment for midwifery practice within a range of settings. Project Report. University of Surrey). Participants detailed their learning activities and support obtained whilst working with their named mentors for approximately 10 days or shifts. The rich audio-diary data have been analysed using Discourse Analysis. A typology of non-formal learning (Eraut, M. 2000. Non formal learning and implicit knowledge in professional work. British Journal of Educational Psychology 70, 113-136) has been used to provide a framework for the analysis. Non-formal learning is defined as any learning which does not take place within a formally organised learning programme (Eraut, M. 2000. Non-formal learning and implicit knowledge in professional work. British Journal of Educational Psychology 70, 113-136). Findings indicate that fear and ambiguity hindered students' learning. Recommendations include the protection of time by mentors within the clinical curriculum to guide and supervise students in both formal and non-formal elements of midwifery practice. This paper will explore the implications of the findings for practice-based education. PMID- 15896417 TI - Evaluation of doctoral nursing programs - a review and a strategy for follow up. AB - Enhancing and assuring the quality of doctoral nursing programs is currently of major concern to promote growth in quality and quantity of research in nursing. The aims of the paper were to review the literature about evaluation of higher education with focus on doctoral programs in nursing, and to present a strategy to evaluate a doctoral nursing program. A search of literature in relevant databases was done using the keywords doctoral program, evaluation, nursing, program evaluation and higher education. From the review it is concluded that more systematic evaluations are necessary to guide the development of quality in nursing. Attention must be given to the curricula, competence in the faculty, the research activity and to the students' involvement in courses and research. Therefore, a strategy for evaluation should be ongoing, flexible, systematic and comprehensive. It should involve students, graduates, employers and faculty members in evaluation, include process and outcome data and give possibility for comparison to internal and external standards. The strategy developed aims to facilitate ongoing and future improvement of the doctoral nursing program. This process is dependent on a methodological pluralism of evaluation to detect what is of most weight for growth in the process of research and education. PMID- 15896418 TI - Patient safety: do nursing and medical curricula address this theme? AB - In this literature review, we examine to what extent patient safety is addressed within medical and nursing curricula. Patient safety is the foundation of healthcare practice and education both in the UK and internationally. Recent research and policy initiatives have highlighted this issue. The paper highlights the significance of this topic as an aspect of study in its own right by examining not only the fiscal but also the human costs such events invite. In the United Kingdom patient safety issues feature prominently in the (Department of Health, 2000a. An organisation with a memory. The report of an expert group on learning from adverse events. The Stationery Office, London, Department of Health, 2000b. Handling complaints: monitoring the NHS complaints procedures (England, Financial year 1998-99). The Stationery Office, London.) policy documentation but this is not reflected within the formal curricula guidelines issued by the NMC and GMC. Yet if healthcare educational curricula were to recognise the value of learning from errors, such events could become part of a wider educational resource enabling both students and facilitators to prevent threats to patient safety. For this reason, the paper attempts to articulate why patient safety should be afforded greater prominence within medical and nursing curricula. We argue that learning how to manage errors effectively would enable trainee practitioners to improve patient care, reduce the burden on an overstretched health care system and engage in dynamic as opposed to defensive practice. PMID- 15896419 TI - Potential role of ubiquinone (coenzyme Q10) in pediatric cardiomyopathy. AB - Pediatric cardiomyopathy (PCM) represents a group of rare and heterogeneous disorders that often results in death. While there is a large body of literature on adult cardiomyopathy, all of the information is not necessarily relevant to children with PCM. About 40% of children who present with symptomatic cardiomyopathy are reported to receive a heart transplant or die within the first two years of life. In spite of some of the advances in the management of PCM, the data shows that the time to transplantation or death has not improved during the past 35 years. Coenzyme Q10 is a vitamin-like nutrient that has a fundamental role in mitochondrial function, especially as it relates to the production of energy (ATP) and also as an antioxidant. Based upon the biochemical rationale and a large body of data on patients with adult cardiomyopathy, heart failure, and mitochondrial diseases with heart involvement, a role for coenzyme Q10 therapy in PCM patients is indicated, and preliminary results are promising. Additional studies on the potential usefulness of coenzyme Q10 supplementation as an adjunct to conventional therapy in PCM, particularly in children with dilated cardiomyopathy, are therefore warranted. PMID- 15896420 TI - Aetiology of inflammatory bowel disease (IBD): role of intestinal microbiota and gut-associated lymphoid tissue immune response. AB - The aetiology of inflammatory bowel disease (IBD) probably involves a combination of genetic predisposition and environmental factors that may be channelled through an abnormality in gut-barrier function, with a loss of antigen tolerance. Some genetic markers that predispose to inflammatory disease have been identified (alleles DR2, DRB1*0103, DRB1*12 and mutations in the NOD2/CARD15 gene on chromosome 16). Alterations in the pattern of cytokine production by T cell subclasses leading to loss of tolerance to oral antigens have been documented. Moreover, a number of environmental factors (cigarette smoking, use of non steroid anti-inflammatory drugs, psychological stress and the presence of the caecal appendix) have been postulated as a trigger of IBD. It has also been suggested that the gut microbiota plays a major role in the development and persistence of IBD, and numerous modifications of intestinal microbiota composition have been identified. As a result, manipulation of the microbiota with antibiotics is a current therapeutic strategy; more recently, however, a number of studies have reported promising results when using probiotic organisms to manipulate gut microbiota composition in order to restore tolerance to microbial antigens of the host's own microbiota. PMID- 15896421 TI - Modulation of nitric oxide and cytokines production by L-arginine in human gut mucosa. AB - BACKGROUND: Arginine is a conditionally essential amino-acid with immuno modulatory properties, mainly through the nitric oxide (NO) pathway. AIM: To assess the effects of arginine on intestinal production of pro- and anti inflammatory cytokines and NO in human gut. METHODS: An enteral solution of arginine or a control solution of amino-acids was administered to 8 healthy volunteers on a randomized cross-over design. Duodenal biopsies were taken. Pro- (IL-6, IL-8) and anti-inflammatory (IL-4, IL-10) cytokines mRNA expression was assessed by RT-PCR. Other biopsies were cultured with 0.1, 0.5 or 2 mM arginine or control amino-acids, under basal or IL-1beta-induced inflammatory conditions. Interleukin-4, IL-6, IL-8 and IL-10 production was measured in culture supernatant by ELISA and NO production by Griess reaction. RESULTS: Arginine enhanced the production of NO under inflammatory conditions in a dose-dependent manner (P=0.03). IL-1beta increased the production of IL-8 and IL-6 (P<0.01). Arginine had no effect on pro- and anti-inflammatory cytokines production both under basal and inflammatory conditions. CONCLUSIONS: Arginine enhanced the production of NO but did not affect that of cytokines in inflammatory human gut. Further clinical studies are required to assess whether arginine-enhanced NO production plays a beneficial or deleterious effect in intestinal inflammation. PMID- 15896422 TI - Total and regional body composition and energy expenditure in multiple symmetric lipomatosis. AB - AIMS: The aim of the present study was to investigate possible alterations in body composition and resting energy expenditure (REE) in type 1 multiple symmetric lipomatosis (MSL). SUBJECTS AND METHODS: Thirteen men aged from 40 to 78 years affected by type I MSL were compared with 13 healthy control subjects. Fat mass (FM) and fat-free mass (FFM) were determined by DEXA using both standard analysis and specifically for the lipomatous region. REE was measured by indirect calorimetry. RESULTS: FM was higher in MSL subjects at proximal arm level, but significantly lower at distal leg level than in controls (left 1.63+/-0.55 vs. 2.26+/-0.49 kg, P<0.05; right 1.63+/-0.53 vs. 2.40+/-0.54 kg, P<0.01). Arm FFM was similar in the two groups, while distal leg FFM was significantly lower in MSL cases (left: 7.8+/-1.3 vs. 8.7+/-0.8 kg, P<0.05; right: 8.0+/-1.5 vs. 9.2+/ 0.9 kg, P<0.05). FFM strongly correlated with REE (r:0.86;P<0.001). REE, expressed as an absolute value and adjusted for FFM (1830+/-215 vs. 1675+/-120 kcal, P<0.05) was higher in MSL patients. CONCLUSION: In conclusion, MSL patients had a marked FFM and FM atrophy in the lower segments of the legs and an altered energy expenditure (hypermetabolism). PMID- 15896423 TI - Does dietary ornithine alpha-ketoglutarate supplementation protect the liver against ischemia-reperfusion injury? AB - Nutritional supplementation with glutamine, arginine and their precursors has been proposed to contribute to the protection against ischemia-reperfusion related injuries. The aim of this study was to evaluate in an isolated perfused rat liver model the preventive effect of a 4-day oral ornithine alpha ketoglutarate (OKG) supplementation against warm ischemia-reperfusion (I-R) injury, and the involvement of nitric oxide synthesis. Rats were fed a controlled regimen supplemented with either OKG (5 g kg(-1); n=15) or an isonitrogenous mixture of non-essential amino acids (Control; n=6) for 4 days. Livers were subsequently prepared for isolated perfusion experiments, including a 45 min no flow ischemic period. The OKG-treated group was divided into two groups according to the absence (OKG; n=8) or presence of a NO-synthase inhibitor, L-N(omega) nitro-arginine methyl ester (OKG L-NAME; n=7) during liver perfusion. Liver cytolysis after ischemia was demonstrated by an elevated alanine aminotransferase release during the last 15 min of reperfusion that was significantly higher in the OKG-L-NAME group. Tumor necrosis factor alpha (TNF(alpha)) production was transiently increased only in the control group just after ischemia. At the end of the reperfusion period, liver superoxide dismutase activity was significantly lower in the OKG-L-NAME group compared to control animals. Dietary OKG administration had only a limited effect in this model of mild hepatic I-R, leading mainly to reduced TNF(alpha) production. As the content of lipid peroxidation products was not modified, it seems that OKG acts on the inflammatory response rather than on oxidative reactions. This action can tentatively be attributed to the role of OKG as a glutamine precursor rather than to the synthesis of arginine and nitric oxide. PMID- 15896424 TI - Oxidant/antioxidant status in liver tissue of vitamin B6 deficient rats. AB - BACKGROUND AND AIMS: The aim of this study was to evaluate the lipid peroxidation and antioxidant function in liver tissue of vitamin B6 deficient rats and investigate relationship among these parameters in either group. METHODS: Twenty four male rats with a weight of 48-59 g were used for the experiment. The rats were divided into control (n=12) and vitamin B6 deficient groups. After 4 weeks of feeding, the rats were killed by cervical dislocation and liver tissues were removed. Biochemical measurements in liver tissue were carried out using a spectrophotometer. RESULTS: Liver tissue antioxidant potential, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, total (enzymatic plus non-enzymatic) superoxide scavenger activity, non-enzymatic superoxide scavenger activity, glutathione levels were significantly lower in vitamin B6 deficient rats than in control group. However, liver tissue glutathione reductase activity, and MDA values were significantly higher in vitamin B6 deficient rats than in control group. CONCLUSIONS: These results explicitly indicate that vitamin B6 deficiency causes a decrease in antioxidant defense system and an increase in oxidant stress in liver tissue in rats. PMID- 15896425 TI - Effect of oral administration of a whole formula diet on nutritional and cognitive status in patients with Alzheimer's disease. AB - AIMS: To evaluate the effect of a whole formula diet on nutritional and cognitive status in Alzheimer's disease patients. METHODS: Patients were randomly assigned to two interventions: a whole formula diet based on lyophilised foods (Treatment Group, n=24) or nutritional advice (Control Group, n=29). Energy intake, body weight, biochemistry, Mini Nutritional Assessment (MNA) and Pfeiffer's tests were determined at baseline and at 3 months of treatment. RESULTS: No differences were observed between groups at baseline. Energy intake tended to increase in the Treatment Group and to decrease in the Control Group, although differences were not significant. The improvement in MNA and Pfeiffer test scores was not significantly different between groups. Body weight increased by 2.06+/-1.9 kg in the Treatment Group and by 0.32+/-3.04 kg in the Control Group (P=0.007). The increases in albumin (P=0.007), haemoglobin (P=0.002) and serum ferritin (P=0.009) were higher in the Treatment Group than in controls. A similar rate of serious adverse events (hospitalisation or death) was observed in both groups. CONCLUSIONS: Administration of this whole formula has a positive impact on nutritional status. The great diversity in textures and tastes enable these formulations to be administered to a wide range of patients with or without liquid dysphagia. PMID- 15896426 TI - Does acute glutamine depletion enhance the response of glutamine synthesis to fasting in muscle in adult and old rats? AB - BACKGROUND AND AIMS: In earlier studies, skeletal muscle glutamine synthetase (GS) activity was shown to be enhanced by fasting and glucocorticoids, and inhibited by exogenous glutamine (Gln) supplementation. The current study was designed to determine whether phenylbutyrate (PhiB), a Gln-chelating agent in humans, (1) could trap Gln and produce a decline in plasma Gln in rats, as it does in humans, and (2) if so, whether (Phi)B would further enhance the response of muscle GS activity to fasting in rats. METHODS: Adult (6-8 months) and aged (20-21 months) rats were fasted for 5 days and received two doses of 0.5 g(Phi)Bby orogastric route at times 0 and 4 h, and were then sacrificed at 5.5 h. Plasma Gln was measured by enzymatic methods, other amino acids were quantified by amino acid analysis. GS activity was measured in soleus (SO) and tibialis anterior (TA) muscles. RESULTS: (Phi)B treatment was associated with: (1) a 20% decline in plasma Gln concentration from 572+/-54 to 424+/-34 micromol/L (P<0.05) and from 476+/-49 to 360+/-80 micromol/L (P<0.05) in fasted adult and old rats, respectively; and (2) a preservation of GS up-regulation by fasting in TA and SO muscles in both adult and aged rats, with TA muscle GS activities of 198+/-65 vs. 203+/-68 ((Phi)B-treated vs. vehicle-treated, NS), and 244+/-81 vs. 274+/-59 (NS) nmol/h/mg protein in adult and aged rats, respectively. CONCLUSION: These data suggest that: (1) large doses of (Phi)B deplete plasma Gln in fasted rats, regardless of age, (2) Gln depletion induced by Phi)B does not alter GS activity. PMID- 15896427 TI - The influence of oxygenated water on the immune status, liver enzymes, and the generation of oxygen radicals: a prospective, randomised, blinded clinical study. AB - BACKGROUND & AIMS: Oxygenated water with an oxygen concentration of 30-120 mg/l water is believed to improve the immune status, without any toxicological effects. The purpose of this clinical study was to assess the effects of long term drinking of oxygenated water on the immune status. METHODS: In this prospective, double-blinded, randomised study 24 volunteers of either sex (age 18 63 years) drank daily 3 times 500 ml either oxygenated (Verum-group: n=12) or normal mineral water (Placebo-group: n=12) for 28 days. On day 1 and day 28 standard laboratory tests, IgG, IgA and IgM, lymphocyte subpopulations and functional analysis of T-cells by flow cytometry, were done. Furthermore, the oxygen radicals were determined by the detection of the ascorbyl radicals. RESULTS: Drinking of normal or oxygenated water had no effect on whole blood count or the liver enzymes. Interestingly the volunteers in the Verum-group showed a significant increase in ascorbyl radicals after drinking oxygenated water for 14 and 21 days. CD4+ and CD4+CD45RA+ lymphocytes as well as lymphocyte activation marker (CD69) and soluble IL-2 receptor increased in both groups, in contrast T-helper2 cells and IgG decreased during the study. The only differences between the two groups were a significant decrease of NK-cells form 13.42%+ or 5.04 to 10.83%+ or -4.82 (P<.002) and an increase of the Th1/Th2-ratio from 2.77%+ or -1.07 to 6.68%+ or -5.33 (P<.03) in the Verum-group. CONCLUSION: Long term consumption of oxygenated water has no apparent harmful effect on the liver, blood and the immune system. Moreover it leads to a transient moderate increase of oxygen radicals in the blood. An interesting observation is the increase of the Th1/Th2-ratio in the Verum group, whereas in both groups T-cell activation after mitogen stimulation, the soluble IL-2 receptor, the CD4+ and the naive CD4+CD45RA+ cells increased. PMID- 15896428 TI - High serum phytosterol levels in short bowel patients on parenteral nutrition support. AB - BACKGROUND & AIMS: Patients with short bowel syndrome (SBS) are often depending on parenteral nutrition support (PNS), sometimes complicated by liver dysfunction. Phytosterols in parenteral lipid emulsions have been suspected to be responsible for cholestasis in paediatric nutrition support. The aim of the present study was to evaluate phytosterol intake and serum phytosterol levels in adult SBS patients. METHODS: We quantified serum levels of phytosterols, cholesterol, and markers for bile acid and cholesterol synthesis, by gas or liquid chromatography in 21 healthy controls, and in 24 adult SBS-patients, 8 with and 16 without PNS. Phytosterols and cholesterol in parenteral lipid emulsions were also quantified. RESULTS: Serum levels in SBS-patients without PNS; with PNS; and in controls, were on average for phytosterols 11; 63; and 23 micromol/l (P<0.05 for differences), cholesterol 4,2; 3,8; and 5,1 mmol/l, lathosterol 808; 824; and 228 micromol/100 mmol cholesterol, and 7alpha-hydroxy-4 cholesten-3-one 207;191; and 18 nmol/l, respectively (P<0.05 between controls and SBS). Phytosterols in lipid emulsions ranged from 591 to 958 micromol/l. CONCLUSIONS: SBS-patients on PNS have higher serum levels of phytosterols than other SBS-patients and controls, possibly because of phytosterols in lipid emulsions. Patients with SBS, regardless of nutrition support, have lower serum levels of cholesterol but higher cholesterol and bile acid synthesis compared to controls. PMID- 15896429 TI - Relationships between structural and functional measures of nutritional status in a normally nourished population. AB - BACKGROUND AND AIMS: Both anthropometric and functional measurements have been used in nutritional assessment and monitoring. Hand dynamometry is a predictor of surgical outcome and peak expiratory flow rate has been used as an index of respiratory muscle function. This study aims to measure in normal subjects the relationship between anthropometric measurements, voluntary muscle strength by hand grip dynamometry and respiratory muscle function by peak expiratory flow rate. METHODS: Ninety-eight subjects (46 male, 52 female) with a mean age of 45.9 years were studied. Hand grip strength was measured in the dominant and non dominant hands with a portable strain-gauge dynamometer. Peak expiratory flow rate was measured using a mini-Wright peak flow meter. Three readings were taken, each 1 min apart, and the average recorded. Midarm muscle circumference (MAMC) was derived from triceps skin fold thickness and midarm circumference (MAC) using standard anthropometric techniques. Statistical relationships were measured with Pearson's coefficient of correlation. RESULTS: In both sexes there was significant correlation between hand grip strength in the dominant and non dominant hands and peak expiratory flow rate (P<0.001). In men, there was a positive correlation between MAMC, hand grip strength (P<0.001) and peak expiratory flow rate (P<0.001). In women muscle function correlated with height (P<0.001) but not MAMC (P>0.05). CONCLUSIONS: In normal subjects bedside tests of skeletal and respiratory muscle function correlated with each other in both sexes, and with muscle mass in men but not in women. PMID- 15896430 TI - Lack of correlation between total lymphocyte count and nutritional status in the elderly. AB - BACKGROUND & AIMS: Malnutrition is a widespread but largely unrecognized problem in aged people. Although absolute total lymphocyte count (TLC) has been proposed as a useful indicator of nutritional status, there is little evidence that low TLC levels reflect malnutrition in the elderly. To examine whether TLC is a suitable marker of malnutrition in the elderly. METHODS: A total of 161 elderly subjects (44 males and 117 females, mean age+/-SD: 77.9+/-7.4; range: 65-95 years) were enrolled from geriatric clinical settings. The participants were categorized according to severely low, low, or normal TLC. Anthropometry measurements, serum albumin, total cholesterol levels, and total score on the mini-nutritional assessment (MNA) were determined. RESULTS: There were no significant differences among the three TLC groups with regard to anthropometry measurements, serum albumin, total cholesterol levels, or MNA score. There was a significant negative correlation of TLC with age, but not with other nutritional markers. The clinical nutritional screening tool, MNA score, was well correlated with all of the nutritional parameters used in the present study except for TLC. CONCLUSION: TLC is not a suitable marker of malnutrition in the elderly. PMID- 15896431 TI - Nutritional support and quality of life in stable chronic obstructive pulmonary disease (COPD) patients. AB - RATIONALE: Nutritional depletion is a common problem in chronic obstructive pulmonary disease (COPD) patients. It is caused, to a large extent, by an imbalance between low-energy intake and high-energy requirements. This problem adversely affects morbidity and mortality. However, the use of nutritional supplements to reach their energy necessities requires optimisation between positive and adverse effects on outcome before being used systematically as part of their comprehensive care. PURPOSE: The aim of our study was to investigate the effects of oral nutritional repletion on quality of life in stable COPD patients. METHODS: Prospective, randomised and multi-centre study. Stable COPD patients with a body mass index 22, a fat-free mass index 16, and/or a recent involuntary weight loss (5% during last month, or 10% during the last 3 months) were studied. Exclusion criteria were to present signs of an airway infection, to have a cardiovascular, neurological, or endocrine disease, to be treated with oral steroids, immunosuppressors or oxygen therapy at home, and to receive nutritional supplements. During 12 weeks, patients were encouraged to ingest a total daily defined energy intake. Randomly, in patients from group A the total daily energy load was Resting Energy Expenditure (REE)x1.7, and those from group B, REE x1.3. Total daily energy intake was achieved with regular food plus, if necessary, oral nutritional supplement rich in proteins (with 50% of whey protein), with predominance of carbohydrates over fat, and enriched in antioxidants. Primary end point variable was quality of life. Secondary end-point outcomes included body weight, body composition, lung function, handgrip strength, and compliance with the energy intake previously planned. Data were treated with a SAS System. Student's test, Wilcoxon's rank sum test, and Mann-Whitney's test were used. RESULTS: At baseline both groups of patients were comparable. All patients needed oral nutritional supplements to achieve total daily defined energy intake. After 12 weeks of follow-up, patients in both groups significantly increased energy intake. Patients in group A increased body weight (P=0.001), triceps skin fold thickness (P=0.009) and body fat mass (P=0.02), and decreased body fat-free mass index (P=0.02). In this group a marked increase in airflow limitation was observed. A tendency to increase body weight and handgrip strength, and to decrease airflow limitation was observed in patients from group B. Furthermore, patients in the later group showed a significant improvement in the feeling of control over the disease (P=0.007) and a tendency to better the other criteria in a quality of life scale. CONCLUSIONS: According to our results, total daily energy intake of REE x 1.3 is preferable to REE x 1.7 in mild stable COPD patients. The administration of oral nutritional supplements, rich in proteins (with 50% of whey protein), with predominance of carbohydrates over fat, and enriched in antioxidants, to achieve total daily defined energy intake in patients in group B was followed by a significant improvement of one criteria (mastery) among many others in a quality of life scale. PMID- 15896432 TI - Dietary treatment of rheumatoid cachexia with beta-hydroxy-beta-methylbutyrate, glutamine and arginine: a randomised controlled trial. AB - BACKGROUND & AIMS: Rheumatoid arthritis (RA) is complicated by cytokine-driven alterations in protein and energy metabolism and consequent muscle wasting (cachexia). The aim of this randomised controlled trial was to investigate the efficacy of a mixture of beta-hydroxy-beta-methylbutyrate, glutamine and arginine (HMB/GLN/ARG) as nutritional treatment for rheumatoid cachexia. METHODS: Forty RA patients supplemented their diet with either HMB/GLN/ARG or a nitrogen (7.19 g/day) and calorie (180 kcal/day) balanced mixture of alanine, glutamic acid, glycine, and serine (placebo) for 12 weeks. Body composition and other outcomes were assessed at baseline and follow-up, and analysed by mixed ANOVA. RESULTS: Dietary supplementation with HMB/GLN/ARG was not superior to placebo in the treatment of rheumatoid cachexia (groupxtime interactions P>0.05 for all outcomes). Both amino acid mixtures significantly increased (main effect of time) fat-free mass (727+/-1186 g, P<0.01), total body protein (719+/-1703 g, P=0.02), arms (112+/-183 g, P<0.01) and legs (283+/-534 g, P<0.01) lean mass, and some measures of physical function. No significant adverse event occurred during the study, but patients in the HMB/GLN/ARG group reported fewer gastrointestinal complaints compared to placebo. CONCLUSIONS: Dietary supplementation with HMB/GLN/ARG is better tolerated but not more effective in reversing cachexia in RA patients compared to the mixture of other non-essential amino acids used as placebo. Further controlled studies are necessary to confirm the beneficial anabolic and functional effects of increased nitrogen intake in this population. PMID- 15896433 TI - A comparison in five European Centres of case mix, clinical management and outcomes following either conventional or fast-track perioperative care in colorectal surgery. AB - BACKGROUND & AIMS: This study reviewed the case mix, clinical management, and clinical outcomes of patients undergoing colorectal resection in five European centres performing different forms of conventional or 'fast-track' perioperative care. METHODS: The perioperative care programme and surgical practice in each centre was defined. Patient data were collected by case-note review on an internet-based audit system. Case mix was determined using ASA classification and the P-POSSUM scoring system. RESULTS: A total of 451 consecutive patients from units practicing either conventional (Sweden, n=109; UK, n=87; Netherlands, n=76, Norway, n=61) or fast-track surgery (Denmark, n=118), were studied between 1998 and 2001. Elements of perioperative practice varied widely both between units practicing 'traditional' care and the reference 'fast-track' unit (Denmark). Based on the P-POSSUM scores, the case mix was similar between centres. There were no differences in morbidity or 30-day mortality between the different centres. The median length of stay was 2 days in Denmark and 7-9 days in the other centres (P<0.05). The readmission rate was 22% in Denmark and 2-16% in the other centres (P<0.05). CONCLUSION: Compared with traditional care, fast-track perioperative care results in a reduced length of hospital stay but may be associated with a higher readmission rate. Morbidity and mortality appears to be similar with either approach. Prospective evaluation of the potential benefits of the fast-track approach in different European centres is merited. PMID- 15896434 TI - Taurolidine lock: the key to prevention of recurrent catheter-related bloodstream infections. AB - The literature shows that repeated courses of antibiotics and catheter removals in a subset of patients suffering from multiple catheter-related bloodstream infections (CRBSI), are unlikely to prevent recurrence. In acceding to preventative strategies, we report our application of the antimicrobial chemotherapeutic Taurolidine used as a daily flush solution in seven home TPN patients suffering from multiple episodes. A pretreatment infection rate of 10.8 infections per 1000 catheter days decreased to 0.8 after treatment. PMID- 15896435 TI - Enhanced recovery after surgery: a consensus review of clinical care for patients undergoing colonic resection. AB - BACKGROUND & AIMS: Clinical care of patients undergoing colonic surgery differs between hospitals and countries. In addition, there is considerable variation in rates of recovery and length of hospital stay following major abdominal surgery. There is a need to develop a consensus on key elements of perioperative care for inclusion in enhanced recovery programmes so that these can be widely adopted and refined further in future clinical trials. METHODS: Medline database was searched for all clinical studies/trials relating to enhanced recovery after colorectal resection. Relevant papers from the reference lists of these articles and from the authors' personal collections were also reviewed. A combination of evidence based and consensus methodology was used to develop the resulting enhanced recovery after surgery (ERAS) clinical care protocol. RESULTS AND CONCLUSIONS: Within traditional perioperative practice there is considerable evidence supporting a range of manoeuvres which, in isolation, may improve individual aspects of recovery after colonic surgery. The present manuscript reviews these issues in detail. There is also growing evidence that an integrated multimodal approach to perioperative care can result in an overall enhancement of recovery. However, effects on major morbidity and mortality remain to be determined. A protocol is presented which is in current use by the ERAS Group and may provide a standard of care against which either current or future novel elements of an enhanced recovery approach can be tested for their effect on outcome. PMID- 15896436 TI - Effective probiotic therapy. PMID- 15896437 TI - Uses of benchmark dose methodology in quantitative risk assessment. PMID- 15896438 TI - Use of mode of action in risk assessment: past, present, and future. AB - The evolution of chemical risk assessment has been marked by a steadily increasing expectation for the use of chemical-specific dosimetric and mechanistic information to tailor the risk assessment approach. The information to be used can range from the broad physical properties of the chemical to detailed information on the mechanism by which it causes a particular toxic outcome, and the risk assessment decisions effected can in turn range from how to define equivalent exposures across species to whether a particular animal outcome is relevant to a human health assessment. A concept that has proven useful in support of these considerations is the "mode of action," a term coined by the USEPA in their new guidelines for carcinogen risk assessment. This paper describes the increasing use of mode-of-action considerations in risk assessment, beginning with early examples involving quantitative dosimetry on the one hand, and qualitative relevance on the other, which foreshadowed the current interest in mode of action. It then describes more recent developments regarding the use of the mode-of-action concept for the selection of a low-dose extrapolation approach, for harmonization of cancer and noncancer risk assessment approaches, and for cross-chemical evaluations. Finally, examples of recent controversies associated with the use of mode-of-action information in risk assessment are provided to demonstrate the challenges that must be overcome to assure the continued viability of the mode-of-action approach. PMID- 15896439 TI - Oral acute toxic class method: a successful alternative to the oral LD50 test. AB - The oral acute toxic class method (ATC method) was developed as an alternative to replace the oral LD50 test. The ATC method is a sequential testing procedure using only three animals of one sex per step at any of the defined dose levels. Depending on the mortality rate three but never more than six animals are used per dose level. This approach results in the reduction of numbers of animals used in comparison to the LD50 test by 40-70%. The principle of the oral ATC method is based on the Probit model and it was first evaluated on a biometric basis before a national and subsequently an international ring study were conducted. The results demonstrated an excellent agreement between the toxicity and the animal numbers predicted biometrically and observed in the validation studies. The oral ATC method was adopted as an official test guideline by OECD in 1996 and was slightly amended in 2001. The ATC method has been successfully used in Germany and in 2003 >85% of all tests on acute oral toxicity testing was conducted as oral ATC tests. In member states of the European Union the ATC method is used in the range of 50% of all tests conducted. Meanwhile the oral LD50 test has been deleted by OECD, by the European Union and by the USA, making the use of alternatives to the oral LD50 test mandatory. PMID- 15896440 TI - Effects of styrene and its metabolites on different lung compartments of the mouse--cell proliferation and histomorphology. AB - Styrene is not carcinogenic in rats but has caused pneumotoxicity and increased lung tumors after inhalation in mice. This study investigated whether styrene-7,8 oxide, ring-oxidized, and side-chain hydroxylated styrene metabolites induce cell proliferation, apoptosis, pathological changes, and glutathione depletion in mice lungs. Intraperitoneal treatment with phenylacetaldehyde and phenylacetic acid (3 x 100 mg/kg b.w./day) increased the levels of apoptosis and cell proliferation in the alveoli without producing any effects in the terminal bronchioli, the target site of tumor formation in mice. Only styrene-oxide (SO) at 3 x 100 mg/kg b.w./day and 4-vinyl-phenol (4-VP) at 3 x 35 and 3 x 20 mg/kg b.w./day, respectively, caused up to 19-fold increases in cell proliferation in the large/medium bronchi and terminal bronchioles; marginal increases in alveolar cell proliferation were noted with SO (1.6-fold) but not with 4-VP. These compounds also caused glutathione depletion in the bronchiolar epithelium and histomorphological changes of the bronchiolar epithelium in large and medium bronchi and terminal bronchioles. Changes were characterized by flattened cells and a loss of the typical bulging of the "dome-shaped" Clara cells, suggesting that Clara cells were primary target cells. The specific reactions of mouse lung to SO and 4-VP could serve as a verifiable hypothesis for the different response of rats and mice with regard to tumor formation. PMID- 15896441 TI - Iodine-deficient vegetarians: a hypothetical perchlorate-susceptible population? AB - Recent risk assessments of environmental perchlorate have been subject to much debate. A particular concern is whether appropriate susceptible sub-populations have been identified. Iodine-deficient pregnant women, especially vegetarians, have been proposed as such a potential susceptible sub-population, but there is no evidence of iodine deficiency in the US population and the adequacy of iodine nutrition has not been studied in US vegetarians. To understand the possibility that US vegetarians might be iodine deficient, we reviewed the prevalence, demography, and lifestyle characteristics of US vegetarians as well as the world literature on iodine nutrition in vegetarians. Our findings indicate that strict vegetarians and vegans, who comprise probably less than 0.1% of the US population, have higher education, higher incomes, and healthier lifestyles than the general population. Field studies indicate that vegetarian diets need not lead to iodine deficiency and vegans may suffer excess iodine intake. It is remains uncertain whether there are iodine-deficient vegans or pregnant women in the US. Of more general concern is whether the 10-fold default uncertainty factor is needed for intraspecies (i.e., within human) variability to protect such hypothetical susceptible sub-populations. PMID- 15896442 TI - The comparative toxicology of 4-chloro-2-methylphenoxyacetic acid and its plant metabolite 4-chloro-2-carboxyphenoxyacetic acid in rats. AB - 4-Chloro-2-carboxyphenoxyacetic acid (CCPA) residues have occasionally been observed in crops treated with 4-chloro-2-methylphenoxyacetic acid (MCPA). The oral toxicity of MCPA and CCPA was compared in a 4-week rat study at a dietary concentration of 2000 ppm. CCPA was also given at 12,000 ppm (equivalent to 1g/kg bodyweight/day). MCPA at 2000 ppm caused reduced food consumption and body weight gain and increased water consumption in females only. Changes in clinical chemistry confirmed the liver as a target organ. Increased serum creatinine and urobilinogen, degenerated transitional epithelial cells in the urine showed that the kidney was also affected. Response to CCPA was confined to the 12,000 ppm dose. The target organs were liver and kidney as for MCPA. Microscopic examination revealed an increased severity of basophilic tubules and calcification at the outer/inner medulla transition in the kidneys. The results demonstrate that CCPA is less toxic than MCPA, that CCPA has no different toxicological end points when compared to MCPA, and that any risks associated with consumption of CCPA will not be underestimated if the CCPA residue is treated as if it were parent MCPA. Based on the MCPA-CCPA comparison, criteria for read across and minimal information requirements are proposed. PMID- 15896443 TI - Lower birth weight as a critical effect of chlorpyrifos: a comparison of human and animal data. AB - Chlorpyrifos is an irreversible inhibitor of cholinesterase (ChE), and inhibition of ChE is believed to be the most sensitive effect in all animal species evaluated and in humans. Recent epidemiology studies reported associations between umbilical cord plasma chlorpyrifos levels and fetal birth weight decreases among minority women living in New York City during pregnancy. These associations raise questions whether impaired fetal development is the critical effect rather than the inhibition of ChE as is currently believed so. We analyze the available information from epidemiology studies and animal studies in order to identify the relative sensitivity of decreased birth weight and inhibition of ChE from exposure to chlorpyrifos. We find that the positive associations from some epidemiology studies are different from other epidemiology investigations. Moreover, a direct comparison of experimental animal neonatal information shows that cholinesterase inhibition is a more sensitive indicator of adverse effect than reduced body weight, and that neonates are equally, or perhaps less sensitive to cholinesterase inhibition than their maternal parent. Based on a review of human studies and comparison of human cord blood chlorpyrifos concentrations with blood chlorpyrifos concentrations that in animals caused effects with good dose-response, it appears unlikely that the exposure level encountered by the population reported in [Whyatt, R.M., Rauh, V., Barr, D.B., Camann, D.E., Andrews, H.F., Garfinkel, R., Hoepner, L.A., Diaz, D., Dietrich, J., Reyes, A., Tang, D., Kinney, P.L., Perera, F.P., 2004. Prenatal insecticide exposures and birth weight and length among an urban minority cohort. Environ. Health Perspect. 112, 1125-1132.] study would cause any fetal developmental effect. Moreover, the critical effect for chlorpyrifos still appears to be cholinesterase inhibition. PMID- 15896444 TI - Comparative carboxylesterase activities in infant and adult liver and their in vitro sensitivity to chlorpyrifos oxon. AB - Maturational expression of carboxylesterase activity in laboratory animals has been correlated with age-related differences in sensitivity to many organophosphorus insecticides including chlorpyrifos. Little information is available, however, on the maturational expression of liver carboxylesterases in humans. Human liver carboxylesterase activity was compared in tissues from infants (2-24 months) and adults (20-36 years). There was no significant difference between mean infant and adult carboxylesterase activities. The carboxylesterase activity rank order was: 2 months<3 months<20 years<24 months<4 months<36 years<21 years<8 months<34 years<35 years. Proteins (3 microg) were separated and blotted using antibodies against rat hydrolase S (HS), human carboxylesterase (HCE) types 1 and 2, and CYP3A4. Again, there were no significant differences in staining density between infant and adult tissues with any isozyme. Aliquots of each sample were pre-incubated (30 min, 37 degrees C) with chlorpyrifos oxon to evaluate in vitro sensitivity. Based on 95% confidence intervals, no significant differences in IC50 values were obtained in 3-month to 36-year samples (range: 1.42-2.12 nM), while the IC50 was significantly lower in the 2-month sample (0.45 nM). Carboxylesterase activity across samples was correlated with cytochrome b5 content and HS immunosignal but not with other microsomal activities (total cyt P450 content, testosterone hydroxylation, coumarin hydroxylation, and EROD). The results suggest that, in contrast to rodents, human liver carboxylesterase expression changes relatively little during postnatal maturation. PMID- 15896445 TI - Assessing prescription medications for priority regulatory review. AB - Poor concordance exists between medications that receive a priority review in Canada and those given an expeditious review in the United States. The objectives of this study were to obtain an evaluation of the clinical significance of new drugs approved in both countries from expert clinical pharmacologists, and to examine the concordance of their aggregate assessment with whether or not the product received an expeditious review in either country. Five experts assessed 146 new medications approved in both Canada and the United States between 1996 and early 2002. Overall, the concordance between the experts' assessments was poor and there was large variation in products considered to be of sufficient importance for priority status. Nevertheless, the experts' evaluations suggested that several priority-reviewed products did not warrant such a review. Regulatory agencies select new medications of potential clinical significance to receive shorter review times to minimize the delay in access to them, but, in Canada, only a low proportion of priority-status products had review times within Health Canada's performance target. The large variation in the assessment of clinical significance suggests that a more appropriate strategy in Canada is to devote sufficient resources to reviewing all medications in a timely manner. PMID- 15896446 TI - The prooxidant effect of sodium metabisulfite in rat liver and kidney. AB - Sodium metabisulfite (Na2S2O5) is used as an antioxidant and antimicrobial agent in a variety of drugs and functions as a preservative in many food preparations. In addition to their antioxidant activity, sulfites oxidize to sulfite radicals (SO3-) initiating lipid peroxidation. This study was performed to elucidate the effect of subchronic Na2S2O5 (520 mg/kg/day) ingestion on hepatic and renal antioxidant enzyme activities and lipid peroxidation in albino rats. The antioxidant effect of l-carnitine was also tested in rats treated with Na2S2O5. Plasma uric acid levels were monitored in all rats included in the study. Malondialdehyde (MDA) levels significantly increased in Na2S2O5 treated rats vs. controls, with kidney values of 2.21+/-0.21 vs. 1.22+/-0.35 and liver values of 79.85+/-19.5 vs. 31.36+/-5.0 nmol/mg protein, respectively. Selenium-glutathione peroxidase (GPx) activity was significantly increased in Na2S2O5 treated rats vs. controls, with kidney values of 38.22+/-2.21 vs. 8.09+/-0.76 and liver values of 31.11+/-6.37 vs. 11.70+/-1.02 U/g protein, respectively. Sodium metabisulfite treatment increased plasma uric acid levels in rats that were included in the study. No protective effect of l-carnitine was observed against lipid peroxidation in both liver and kidneys of rats treated with Na2S2O5. The presented data confirm the prooxidant activity of sulfites and suggest that increased GPx activity and plasma uric acid levels may partially reduce the observed renal and hepatocellular oxidative damage caused via the ingestion of sulfites. PMID- 15896447 TI - Risk assessment of chemicals and pharmaceuticals in the pediatric population: a workshop report. AB - ATSDR and RIVM organized an Expert Panel Workshop on the Differences Between Children and Adults and Their Relevance to Risk Assessment. The workshop was held in June 2003, in Brussels, Belgium. The purpose of the workshop was to identify data gaps in current scientific knowledge related to children's health and to recognize areas of mutual interest that would serve as the basis for upcoming ATSDR/RIVM cooperative projects. The aim for both agencies is a better understanding of the issues related to children's health, and the improvement of scientifically based (chemical) risk assessment in children. Topics discussed included clinical trials/toxicity studies, testing in juvenile animals, PBPK modeling in children, and children's risk assessment. PMID- 15896448 TI - Food packaging regulation in the United States and the European Union. AB - The regulation of food packaging has, indeed, become a global subject as the world's commercial channels have broadened since World War II. The first comprehensive regulatory statute governing the area was the Food Additives Amendment of 1958 in the United States, a modification of the 1938 Federal Food, Drug, and Cosmetic Act. Germany, Italy, The Netherlands, and Belgium began regulatory activity in the early 1960s and the European Community set in motion its own work in this direction beginning in 1976. Companies in the United States and the Europe have been struggling with each of the laws since then. The regulatory systems employed on the two continents are superficially similar but this is far from the truth in actual practice. Each has its own special history and set of exemptions; they vary tremendously. Each has its cadre of supporters as well but a considerable lack of understanding afflicts regulators and the regulated on both sides of the Atlantic. This three part report first discusses the history of the United States system and its characteristics; Book II covers the history of European Union regulations and their salient features; and Book III highlights the commonalities and differences in the systems, discusses their scientific basing points, and makes some recommendations as to how they can be brought closer together in the interests of harmonization and the removal of unnecessary trade barriers. PMID- 15896449 TI - A review and critique of the EPA's rationale for a fine particle standard. AB - I review the rationale for the Environmental Protection Agency's 1996 fine particle standard, which was based almost entirely on the epidemiological data with neither support from Toxicology nor understanding of mechanism. While many epidemiological papers available in 1996 reported associations between ambient particles and adverse effects on human health, many others did not and the evidence fell far short of supporting a causal association between particle mass concentration and human health. The literature appearing after 1996 further complicates the picture. The large studies that have appeared after 1996, such as National Mortality Morbidity and Air Pollution Study, and the reanalyses of the American Cancer Society II study, report risks that are substantially smaller than the risks reported in the 1996 Criteria Document and Staff Paper. Moreover, concerns about confounding by weather, temporal trends and co-pollutants remain unresolved. Other issues having to do with model choice have resurfaced as a result of reanalyses of critical data to address a glitch in a widely used software package for time-series epidemiology studies of air pollution. Finally, contemporary examples show that the results of observational epidemiology studies can be seriously biased, particularly when estimated risks are small, as is the case with studies of air pollution. The Agency has largely ignored these issues. I conclude that a particle mass standard is not defensible on the basis of a causal association between ambient particle mass and adverse effects on human health. Such a standard may be justifiable on the basis of the precautionary principle, however. The Agency could argue that the Science raises concerns about current levels of air pollution, and that reduction of ambient fine particulate matter mass, if it could be achieved without an increase in the level of the ultrafines, could have positive effects on human health. If the Agency justifies a particulate matter mass standard on these grounds then the debate over the form and level of the standard will, for all practical purposes, belong strictly in the Policy arena. PMID- 15896450 TI - Human health risks from exposures to perfluorooctanoic acid: a critique of Butenhoff et al. 2004. PMID- 15896452 TI - Chemopreventive and therapeutic effects of curcumin. AB - Chemoprevention is a promising anti-cancer approach with reduced secondary effects in comparison to classical chemotherapy. Curcumin, one of the most studied chemopreventive agents, is a natural compound extracted from Curcuma longa L. that allows suppression, retardation or inversion of carcinogenesis. Curcumin is also described as an anti-tumoral, anti-oxidant and anti-inflammatory agent capable of inducing apoptosis in numerous cellular systems. In this review, we describe both properties and mode of action of curcumin on carcinogenesis, gene expression mechanisms and drug metabolism. PMID- 15896453 TI - Involvement of PKC and ROS in the cytotoxic mechanism of anti-leukemic decursin and its derivatives and their structure-activity relationship in human K562 erythroleukemia and U937 myeloleukemia cells. AB - Protein kinase C (PKC) plays an important role in the proliferation and differentiation of various cell types including normal and leukemic hematopoietic cells. Recently, various PKC modulators were used as a chemotherapeutic agent of leukemia. Decursin (1), a pyranocoumarin from Angelica gigas, exhibits the cytotoxic effects on various human cancer cell lines and in vitro PKC activation. For the development of more effective anticancer agents with PKC modulation activity, 11 decursin derivatives 2-12 were chemically synthesized and evaluated for their ability to act as a tumor-suppressing PKC activator and as an antagonist to phorbol 12-myristate 13-acetate (PMA), a tumor-promoting PKC activator. In the presence of phosphatidylserine (PS), all of 12 compounds 1-12 activated PKC (mainly alpha, beta, and gamma isozymes) but only three compounds 1 3 activated PKC even in the absence of PS. Six compounds 1-6 containing the coumarin structure were cytotoxic to human K562 erythroleukemia and U937 myeloleukemia cells. A cytotoxic mechanism of decursin and its derivatives was investigated using TUR cells, a PKC betaII-deficient variant of U937 cells. Among six compounds 1-6 with cytotoxicity to K562 and U937 leukemia cells, only three compounds 1-3 were cytotoxic to TUR cells. Therefore, compounds 1-3 and 4-6 inhibit the proliferation of leukemia cells in a PKC betaII-independent and dependent manner, respectively, indicating that the side chain of compounds determines the dependency of their cytotoxicity on PKC betaII. To further elucidate the cytotoxic mechanism of compounds 1 and 2, levels of PKC isozymes and generation of reactive oxygen species (ROS) were investigated. Compounds 1-2 induced the down-regulation of PKC alpha and betaII in K562 cells and the production of ROS in U937 cells. Thus, PKC and ROS are probably important factors in the cytotoxic mechanism of compounds 1-2. From these results, the structure activity relationship of decursin and its derivatives is as follows: (i) the coumarin structure is required for anti-leukemic activity and (ii) the side chain is a determinant of PKC activation and the cytotoxic mechanism in leukemia cells. PMID- 15896454 TI - Polyethylene glycol reduces inflammation and aberrant crypt foci in carcinogen initiated rats. AB - Polyethylene glycol 8000 inhibits the formation of tumors and of aberrant crypt foci (ACF) in carcinogen-initiated rats. We asked: is the inhibition associated with a reduction of colonic inflammation and an increase in colonic cell permeability? Twenty-eight, male F 344 rats were divided into two groups, 10 control animals and 18 animals initiated with azoxymethane. Nine of the rats in the carcinogen-initiated group were given a diet with 5% PEG 8000 in an AIN-93 based, high fat diet. The other nine, and the control group received the diet without the addition of PEG. Nine weeks later, the rats receiving the diet containing PEG had a 43% reduction in ACF (P<0.001) compared with the carcinogen initiated rats on the control diet, a result confirming earlier observations that PEG inhibits colon carcinogenesis. The animals receiving the diet containing PEG also had a 10-fold reduction in fecal granulocyte marker protein (GMP) (P<0.001) compared with both the carcinogen-treated and the control animals. PEG reduced inflammation below the levels of carcinogen-treated and of untreated animals. Fecal water from the rats receiving PEG did not reduce transepithelial resistance of, or manitol flux through, human Caco-cells grown as monolayers in vitro. PEG may reduce colon carcinogenesis through a mechanism involving colonic inflammation. PMID- 15896455 TI - Actinonin induces apoptosis in U937 leukemia cells. AB - We have previously shown that actinonin causes inhibition of cellular proliferation in U937 leukemia cells. In this report we demonstrate that the inhibition of cell growth by actinonin occurs through the induction of apoptosis. Signs of apoptosis at high actinonin concentration included DNA fragmentation, exposure of phosphatidylserine and condensation of cell nuclei. Apoptosis caused by actinonin was inhibited by Z-VAD-FMK, a broad specificity inhibitor of caspases, implicating the caspase pathway of apoptosis. Further, apoptosis was associated with a large increase in intracellular caspase-3 and -7 activities. PMID- 15896456 TI - The DNA repair gene ERCC2/XPD polymorphism Arg 156Arg (A22541C) and risk of lung cancer in a Chinese population. AB - To determine the effect of the DNA repair gene XPD Arg156Arg polymorphism on the risk of lung cancer in a North-Eastern Chinese population, a hospital-based case control study was designed consisting of 149 newly diagnosis subjects with lung cancer and 137 cancer-free control subjects matched on age (+/-3 years), gender and ethnicity. In the whole study group, XPD Arg156Arg was not associated with risk of lung cancer. In stratified analyses, the variant A-allele of XPD Arg156Arg was associated with increased risk of adenocarcinoma of lung (AA/AC versus CC; adjusted OR=1.65; 95% CI=1.09-2.50) (P=0.02). Furthermore, the presence of one or two variant A-alleles was associated with increased risk for lung cancer (OR=2.49; 95% CI=1.10-5.64) (P=0.03) and adenocarcinoma of lung (OR=5.60; 95% CI=1.52-20.56) (P=0.005) among never-smokers only. These results suggest a possible gene-environment interaction. This is the first study to report a significant association of XPD Arg156Arg with risk of lung cancer. PMID- 15896457 TI - Synergistic interaction between platinum-based antitumor agents and demethylcantharidin. AB - A novel series of TCM-platinum complexes [Pt(C8H8O5)(NH2R)2] 1-5, designed from incorporating demethylcantharidin, a modified component from a traditional Chinese medicine (TCM) with a platinum moiety was found to circumvent cisplatin resistance in mouse leukemia and human hepatocellular carcinoma. These properties are most likely due to the inclusion of the protein phosphatase 2A (PP2A) inhibiting demethylcantharidin in the novel compounds. We have investigated the potential synergistic effect of combining demethylcantharidin with a platinum based antitumor agent, such as cisplatin, carboplatin, or oxaliplatin in vitro against L1210 mouse leukemia and SK-Hep-1 human hepatocellular carcinoma, and in vivo against a SK-Hep-1 subcutaneous-inoculated xenograft in nude mice, using median effect analysis. Demethylcantharidin and the platinum antitumor agents were synergistic in all cell lines tested in vitro, and the most effective antiproliferative regimen was when demethylcantharidin was added 24 h before cisplatin. Synergistic antitumor activity was also demonstrated in vivo without undue toxicity; no excessive loss in mouse body weight or overt pathology were observed at the effective doses. The results support a new approach for augmenting cytotoxic effect of established Pt-based drugs with demethylcantharidin in treating human hepatocellular carcinoma and other solid tumors. PMID- 15896458 TI - Induction of apoptosis in human leukemia cells by MCS-C2 via caspase-dependent Bid cleavage and cytochrome c release. AB - The purpose of the present study was to investigate the anti-proliferative and apoptotic effects of MCS-C2, a novel synthetic analogue of the pyrrolo[2,3 d]pyrimidine nucleoside toyocamycin and sangivamycin, in human promyelocytic leukemia (HL-60) cells. When treated with 5 microM MCS-C2, inhibited proliferation associated with apoptotic induction was found in the HL-60 cells in a concentration-dependent and time-dependent manner, plus nuclear DAPI staining revealed the typical nuclear features of apoptosis. However, MCS-C2 showed almost no antiproliferative effect and no apoptotic induction in normal lymphocyte cells used as a control when compared with those in HL-60 cancer cells. Moreover, a flow cytometric analysis of the HL-60 cells using FITC-dUTP and propidium iodide (PI) showed that the apoptotic cell population increased gradually from <1% at 0 h to 34% at 12 h after exposure to 5 microM MCS-C2. This apoptotic induction was associated with the cleavage of Bid and a release of cytochrome c from mitochondria into the cytosol, followed by the activation of caspase-3 and inactivation of poly(ADP-ribose) polymerase (PARP). However, there was no significant change in any other mitochondrial membrane proteins, such as Bcl-2 and Bax. Consequently, the current findings suggest that the mitochondrial pathway was primarily involved in the MCS-C2-induced apoptosis in the human promyelocytic leukemia HL-60 cells. PMID- 15896459 TI - Etoposide (VP-16) elicits apoptosis following prolonged G2-M cell arrest in p53 mutated human non-small cell lung cancer cells. AB - In this work, we described the proliferation of human non-small-cell-lung-cancer (NSCLC) cells H1437 harboring p53 alleles (proline-267) can be inhibited by low dosage topoisomerase II inhibitor etoposide (VP-16) in vitro and in vivo. The cytotoxicity was demonstrated by prolonged cell arrest at G2-M checkpoint exhibiting senescence-like phenotype followed by apoptotic cell death that appeared on the sixth day of VP-16 treatment. The experimental in vivo evidence of growth suppression was also demonstrated in xenograft tumors. The appearance of senescence-like state during extended G2-M phase arrest was indicated by slow proliferation and loss of growth sensitivity in culture accompanied with cellular morphological changes, time-dependent regulation of beta-galactosidase staining as well as distinct reduction of telomerase activity upon protracted VP-16 exposure. Further molecular determinants leading to G2-M cell arrest was also characterized by the concerted up-regulation of cyclin-dependent kinase inhibitors, p16(INK4a) and p21(Waf1/Cipi), beginning 2 days later following drug exposure at both translational and transcriptional levels, while human telomerase reverse transcriptase (hTERT) activities reduced progressively. The clinically important therapeutic agent VP-16-mediated prolonged cell arrest at G2-M phase prior to apoptotic death offered a different perspective in restraining human cancer cells at low drug dosage, thereby serving as an effective telomerase inhibitor as well as an apoptosis effector. The overall results demonstrated that apoptosis can be regulated differently in human NSCLC cells with disrupted p53. Further effort in elucidating G2-M arrest before leading to apoptosis promises to provide an alternative insight in reversing tumorigenic phenotype of human cancers. PMID- 15896460 TI - Cyclin D1 G870A polymorphism and squamous cell carcinoma of the uterine cervix in Korean women. AB - Though many investigators have reported relationships between the CCND1 polymorphism and susceptibility to various carcinomas, to our knowledge, no report has been issued concerning its relationship with uterine cervical cancer. Thus, we undertook this study to investigate the association between CCND1 polymorphisms and susceptibility to cervical cancer in Korean women. This study was carried on 222 patients with squamous cell carcinoma of uterine cervix and on 314 normal controls. CCND1 genotyping was determined by polymerase chain reaction and restriction fragment length polymorphism. The allelic frequencies of the cases (A, 0.53; G, 0.47) were not significantly different from those of the controls (A, 0.49; G, 0.51) (P=0.238). Regression analysis after adjusting for age showed that the CCND1 G870A genotypes are not related to the risk of squamous cell carcinoma of the uterine cervix. Our findings suggest that the CCND1 polymorphism is not associated with an increased risk of squamous cell carcinoma of uterine cervix in Korean women. PMID- 15896461 TI - Rapid detection of single nucleotide polymorphisms related with lung cancer susceptibility of Chinese population. AB - Genetic variations have been thought to contribute to individual differences in lung cancer susceptibility. In our study, the possibility of an association of CYP1B1, GSTP1 and hOGG1 genetic polymorphisms with lung cancer was investigated in Chinese population of Nanjing, by a new single nucleotide polymorphism (SNP) typing approach of di-allele-specific-amplification with artificially modified primers (diASA-AMP) technique. A matched case-control study of 227 patients with lung cancer was conducted to detect CYP1B1 Leu432Val, GSTP1 Ile105Val and hOGG1 Ser326Cys polymorphisms. Genotypes were analyzed by diASA-AMP technique. Results did not show a significant difference in distributions of allele frequencies or genotypes of CYP1B1, GSTP1 and hOGG1 between two groups. However, stratifying on smoking status demonstrated that CYP1B1 432Val genotype had a slightly combined effect on lung cancer with smoker subjects (OR=2.78, 95%CI=1.46-5.29). The interaction between GSTP1 105Val mutation and smoking in the development of lung cancer were not detected, nor was hOGG1 326Cys mutation. Variant allele frequencies of CYP1B1, GSTP1 and hOGG1 in control group were similar to other reports of Chinese population. The sequencing results of CYP1B1, GSTP1 and hOGG1 matched the ones of diASA-AMP technique. CYP1B1 432Val polymorphism may modulate the individual susceptibility of lung cancer among smokers in Chinese population. GSTP1 Ile105Val and hOGG1 Ser326Cys polymorphisms were not found to be risk factors of lung cancer in this study. The method diASA-AMP is rapid, specific and cost-effective. It can be used for rapid detection of the genes related with tumor susceptibility of population. PMID- 15896462 TI - UV induced bystander signaling leading to apoptosis. AB - Human keratinocytes (HaCaT) were exposed to UV (A+B) (UVA-350-400 mJ/cm2 and UVB 30 mJ/cm2) which induces apoptosis as evidenced by MTT assay, DNA laddering, Bax and Fas up-regulation. UV induced apoptotic conditioned media (6 h or earlier) did not cause apoptosis in unexposed cells. However, treatment with conditioned medium collected post UV exposure (1 h) induced Bax in unexposed cells as observed by RT-PCR. The induction of cell death was initiated by conditioned medium collected 12 h after UV exposure and the extent of death was increased progressively when conditioned medium collected 24 or 72 h post UV exposure was used. Medium collected 24 h after UV exposure also increased mitochondrial membrane permeability as determined by rhodamine uptake. Conditioned medium induced apoptosis did not involve reactive oxygen species (ROS) unlike UV induced apoptosis indicating that the apoptosis pathway could be different. Interestingly, at high dilution apototic conditioned medium did not induce apoptosis but actually protected cells from UV insult. The role of nerve growth factor (NGF) in UV induced bystander effects are also discussed. PMID- 15896463 TI - A novel missense germline mutation in exon 2 of the hMSH2 gene in a HNPCC family from Southern Italy. AB - Germline mutations within the mismatch repair (MMR) genes are generally found in colorectal cancer (CRC) patients with a positive family history for the presence of the neoplasia. Clinical standard criteria have been established to define hereditary-non-polyposis-colorectal cancer (HNPCC)-prone families. Interestingly, the number of MMR gene mutations found in kindreds not fulfilling these criteria is still increasing. In this work we report the identification of a novel germline mutation of the hMSH2 gene, in two CRC-bearing subjects. The two probands belong to a large kindred from South Italy with no history suggestive for cancer aggregation. On the other hand, the early-onset of the neoplasia as well as the presence of a high number of tumor infiltrating lymphocytes (TILs) in the histological specimens of both patients, prompted us to perform a comprehensive genetic analysis. This analysis included the evaluation of the microsatellite instability (MSI) status with five markers according to the National Cancer Institute recommendations, followed by direct sequencing of the hMLH1 and hMSH2 genes. Both probands were found to carry a germline missense (277 C>T) mutation leading to the change (L93F) of an amino acid residue in a highly conserved domain of the MSH2 protein. This mutation is accompanied by the loss of expression of the hMSH2 gene in the tumor tissue. Our findings suggest that in the presence of the above-mentioned criteria it may be useful to perform a molecular analysis of the MMR genes, even if the pedigree does not show marked aggregation of cancers. PMID- 15896464 TI - The mechanism of ellipticine-induced apoptosis and cell cycle arrest in human breast MCF-7 cancer cells. AB - Ellipticine, a cytotoxic plant alkaloid, is known to inhibit topoisomerase II. Here, we first report the molecular mechanism of ellipticine's apoptotic action in human breast MCF-7 cancer cells. Treatment of cells with ellipticine resulted in inhibition of growth, and G2/M phase arrest of the cell cycle. This effect was associated with a marked increase in the protein expression of p53 and, p21/WAF1 and KIP1/p27, but not of WAF1/p21. Ellipticine treatment increased the expression of Fas/APO-1 and its ligands, mFas ligand and sFas ligand, and subsequent activation of caspase-8. The mitochondrial apoptotic pathway amplified the Fas/Fas ligand death receptor pathway by Bid interaction. This effect was found to result in a significant increase in activation of caspase-9. Taken together, we have concluded that the molecular mechanisms during ellipticine-mediated growth inhibition and induction of apoptosis in MCF-7 cells were due to (1) cell cycle arrest and induction of apoptosis, (2) induction of p53 and KIP1/p27 expression, (3) triggering of Fas/Fas ligand pathway, (4) disruption of mitochondrial function, and (5) the apoptotic signaling was amplified by cross talk between Fas death receptor and mitochondrial apoptotic pathway. PMID- 15896465 TI - Functional definition of relevant epitopes on the tumor suppressor PTEN protein. AB - The binding of PTEN to PDZ-domain-containing proteins appears to play an important role in the control of cell growth, motility and apoptosis. In turn, this binding can be abrogated by cleavage of the PTEN C-terminal region by caspase-3. We have generated and characterized monoclonal antibodies (mAb) directed against distinct epitopes at the C-terminal region of PTEN, and used them to define protein-binding epitopes on PTEN and to study its cleavage by caspase-3. mAb directed against epitopes at the far C-terminus of PTEN blocked binding to PTEN cognate PDZ domains and did not recognize the caspase-3 cleaved PTEN fragments. On the other hand, mAb that recognized an epitope within the C2 domain of PTEN did not prevent binding to PDZ domains, but could detect the caspase-3 cleaved PTEN fragments. The analysis of PTEN cleavage by caspase-3 revealed that the lipid phosphatase activity of PTEN controls its own degradation by interfering with the PI3-K anti-apoptotic activity. Our results define protein binding sites on the PTEN tumor suppressor at the immunochemical level, and suggest a regulatory link between PTEN phosphatase activity, caspase-3 sensitivity and PTEN-protein interactions. PMID- 15896466 TI - Interactions between human colon carcinoma cells, fibroblasts and monocytic cells in coculture--regulation of cathepsin B expression and invasiveness. AB - To elucidate the importance of the tumor/host interaction in malignant tumors, we investigated the colon carcinoma cell line HT-29 in coculture with monocytic cells (THP-1) and fibroblasts (175BR) for cathepsin B expression and activity. The tumor cells were grown in monolayer cultures or as multicellular tumor spheroids. After coculture, the three cell types were separated by labeled magnetic beads for cathepsin B mRNA and protein analysis. The invasive potential was studied in in vitro invasion assays. The expression level of cathepsin B was found to be 10-fold increased in three dimensional spheroids of HT-29 compared to HT-29 monolayers. The coculture of HT-29 with THP-1 cells and/or human fibroblasts led to a considerable increase in cathepsin B mRNA expression in both tumor and tumor-associated cells. The invasive potential of the tumor cells was 5 times increased by adding monocytic cells to the assay system. This is dependent on the functional activity of cathepsin B as shown by specific siRNA's and seems to be regulated by activation of ERK1/2 and p38 signal transduction pathways. PMID- 15896467 TI - Tryptophan degradation in patients with gynecological cancer correlates with immune activation. AB - Tryptophan degradation by the enzyme indoleamine-(2,3)-dioxy genase (IDO) and neopterin production are induced within cellular immune activation by stimulation of monocyte-derived macrophages and dendritic cells with cytokine interferon gamma. Deprivation of tryptophan represents an important antimicrobial and antitumoral immune defence mechanism but it also suppresses T-cell proliferation. Recently tryptophan degradation by tumor cells was proposed as strategy to escape immune response. In this study the relationship between tryptophan degradation and immune activation was examined in 20 patients with gynecological cancer. Concentrations of tryptophan and kynurenine were measured by HPLC in sera of patients, and to estimate IDO activity, the kynurenine to tryptophan ratio was calculated. In parallel, neopterin concentrations were measured by ELISA. Tryptophan concentrations (median, interquartile range: 43.5, 31.2-56.3 microM) were lower in patients with gynecological cancer compared to healthy individuals of similar age (53.5, 47.0-64.2 microM; P<0.05). Kynurenine concentrations (median: 1.91 vs. 1.73 microM in controls) and kyn/trp (median: 41 vs. 35 micromol/mmol in controls) were slightly higher in patients, but not significantly different. Neopterin concentrations were significantly higher in patients (median: 10.8 vs. 7.0 nM in controls; P<0.05) and correlated with the kynurenine per tryptophan ratio (r(s)=0.555; P<0.02). In conclusion, tryptophan degradation is detectable in patients with gynecological cancer. The relationship between kyn/trp and neopterin concentrations indicates that cellular immune activation rather than tumor-mediated IDO-activity is responsible (228 words). PMID- 15896468 TI - The inhibitor of apoptosis (IAP) survivin is expressed in human testicular germ cell tumors and normal testes. AB - Deregulated apoptosis of germ cells may contribute to malignant transformation as well as male infertility. We analyzed expression of the inhibitor of apoptosis survivin by polymerase chain reaction (PCR) in testicular germ cell tumors (TGCTs, n=28), normal testes (n=19) and testes with defective spermatogenesis (n=22). In a subset of samples (n=35), survivin transcript levels were quantified using real-time PCR. While survivin mRNA was expressed at high levels in undifferentiated TGCTs and normal testes, it was lost in mature teratomas and germ cell aplasia. Survivin expression in both normal and transformed germ cells is in contrast to its sharp differential expression in other tissues. Survivin expression correlates with the differentiation state of TGCTs and may be relevant for their tumorigenesis and malignant potential. PMID- 15896469 TI - Correlation of Wnt-2 expression and beta-catenin intracellular accumulation in Chinese gastric cancers: relevance with tumour dissemination. AB - Wnt/beta-catenin signalling pathway is integrally associated with human tumour development and progression. Aberrant beta-catenin intracellular distribution has been found in gastric cancer, but the pattern of Wnt expression in stepwise gastrocarcinogenesis and its potential influence in beta-catenin distribution are still lesser known. By the methods of frozen tissue array-based immunohistochemistry, Western blot analysis and RT-PCR, a paralleled study was conducted to check Wnt2 expression and beta-catenin intracellular distribution in two major subtypes of gastric cancers (intestinal gastric cancer, i-GC and diffuse gastric cancer, d-GC) and their premalignant (intestinal metaplasia, IM and chronic gastritis, CG) and noncancerous counterparts. According to the results obtained and the clinical data collected, correlation of Wnt2 expression with beta-catenin translocalisation and their links with tumour dissemination were elucidated. The results demonstrated (1) that Wnt2 expression and cytoplasmic/nuclear beta-catenin accumulations appeared in most gastric cancers irrespective to their morphological phenotypes, (2) that over-expressed Wnt and nuclear translocalisation of beta-catenin were found in 68 and 58% of i-GCs and in 47 and 47% of d-GCs in a closely related pattern (P<0.01) and (3) that co existence of Wnt2 up-regulation/beta-catenin nuclear translocalisation were positively associated with lymph node metastasis (P<0.05) as well as T-stage. These data indicate that Wnt/beta-catenin signalling pathway is activated in most of gastric cancers, which may play pivotal roles either in gastric cancer formation or in tumour invasion and dissemination. PMID- 15896470 TI - Botulinum toxin: more than meets the naked eye! PMID- 15896471 TI - Gene expression of myogenic regulatory factors following intramuscular injection of botulinum A toxin in juvenile rats. PMID- 15896472 TI - Prenatal morphine exposure affects sympathoadrenal axis activity and serotonin metabolism in adult male rats both under basal conditions and after an ether inhalation stress. AB - We have previously shown that prenatal morphine exposure inhibited the hypothalamo-pituitary-adrenal (HPA) axis and altered the hypothalamic metabolism of serotonin during the early postnatal period in the rat and induced a chronic sympathoadrenal hyperactivity under resting conditions in adult male rats. In this study, we examined the effects of prenatal morphine exposure on the responsiveness to an acute ether inhalation stress of the sympathoadrenal and HPA axis and the hippocampal and hypothalamic concentrations of serotonin (5HT) and 5 hydroxylindoleacetic acid (5HIAA) in 3-month-old male rats. The plasma levels of adrenocorticopic hormone (ACTH) and corticosterone (B) did not differ between the two groups both under resting conditions and after ether exposure. Ether inhalation increased adrenal tyrosine hydroxylase (TH) and phenylethanolamine N methyltransferase (PNMT) mRNA expression as well as adrenal epinephrine (E) concentration in control rats but not in prenatally morphine-exposed (PM) animals. Under basal conditions, hypothalamic concentrations of 5HT and 5HIAA increased in PM animals. In contrast to control animals, PM rats showed, in response to stress, an increased level of 5HT and 5HIAA in both the hypothalamus and in the hippocampus. In conclusion, prenatal morphine exposure produces long lasting alterations in brain serotonin transmission and in the sympathoadrenal responsiveness to an acute systemic stress. PMID- 15896473 TI - Audiovisual prior entry. AB - It is almost one hundred years since Titchener [E.B. Titchener, Lectures on the Elementary Psychology of Feeling and Attention, Macmillan, New York, 1908] published his influential claim that attending to a particular sensory modality (or location) can speed up the relative time of arrival of stimuli presented in that modality (or location). However, the evidence supporting the existence of prior entry is, to date, mixed. In the present study, we used an audiovisual simultaneity judgment task in an attempt to circumvent the potential methodological confounds inherent in previous research in this area. Participants made simultaneous versus successive judgment responses regarding pairs of auditory and visual stimuli at varying stimulus onset asynchronies (SOAs) using the method of constant stimuli. In different blocks of trials, the participants were instructed to attend either to the auditory or to the visual modality, or else to divide their attention equally between the two modalities. The probability of trials containing intramodal stimulus pairs (e.g., vision-vision or audition-audition) was increased in the focused attention blocks to encourage participants to follow the instructions. The perception of simultaneity was modulated by this attentional manipulation: Visual stimuli had to lead auditory stimuli by a significantly smaller interval for simultaneity to be perceived when attention was directed to vision than when it was directed to audition. These results provide the first unequivocal evidence for the existence of audiovisual prior entry. PMID- 15896474 TI - Kainic acid (KA)-induced Ca2+/calmodulin-dependent protein kinase II (CaMK II) expression in the neurons, astrocytes and microglia of the mouse hippocampal CA3 region, and the phosphorylated CaMK II only in the hippocampal neurons. AB - In the present study, we investigated the role of Ca2+/calmodulin-dependent protein kinase II (CaMK II) and which types of neuronal cells contain CaMK II and phosphorylated CaMK II (p-CaMK II) in the CA3 hippocampal region of mice using confocal immunofluorescence study. KA increased the CaMK II, p-CaMK II, glial fibrillary acidic protein (GFAP) and complement receptor type 3 (OX-42) immunoreactivities (IR) at 30 min after KA treatment in mouse hippocampal area. In studies, nevertheless KA-induced CaMK II is expressed in neurons or astrocytes or microglia, p-CaMK II is expressed only in neurons. Thus, our results suggest that the activated CaMK II in early time may be performed important roles only in neurons but not in the astrocytes and microglia. PMID- 15896475 TI - Neuroglial activation in Niemann-Pick Type C mice is suppressed by intracerebral transplantation of bone marrow-derived mesenchymal stem cells. AB - Glial activation is thought to play a key role in pathogenesis of neurodegenerative disorders. Here we show that direct transplantation of bone marrow-derived mesenchymal stem cells (BM-MSC) results in alleviation of inflammatory responses associated with the cerebellum of Niemann-Pick disease Type C (NP-C) model mice. Immunohistochemical examinations using glial fibrillary acidic protein (GFAP) and F4/80 antibodies revealed that BM-MSC transplantation reduced significantly both of astrocytic and microglial activations in the cerebellum of NP-C mice. Expression of macrophage colony stimulating factor (M CSF), a microglial activator, was also considerably down-regulated by the BM-MSC transplantation. These findings suggest that BM-MSC transplantation may have potential for a therapeutic role in the treatment of NP-C and other neurodegenerative brain disorders. PMID- 15896476 TI - Sex difference in the distribution and size of glomeruli in the ferret's main olfactory bulb. AB - When exposed to male anal scent gland odorants in a previous study from our laboratory, the distribution of activated glomeruli in the ventral-caudal portion of the main olfactory bulb (MOB) was greater in female than in male ferrets. We asked whether this functional dimorphism corresponds to a morphological sex difference in the distribution, number, or size of glomeruli in the MOB of adult ferrets. Coronal serial sections through the rostro-caudal extent of the MOB from groups of breeding male and female ferrets were collected, and the glomeruli were visualized after staining of juxtaglomerular cells with an antiserum raised against neuronal nuclear protein. In both sexes the greatest density of glomeruli was seen in the ventral MOB; however, this dense cluster of glomeruli extended more caudally in males than in females. Also, the number of glomeruli per section across the caudal extent of the MOB and glomerular areas measured at three sites in the MOB were significantly greater in males than in females. We previously observed greater odor-induced glomerular activation in the ventral-caudal MOB of female than male ferrets. This functional sex difference was inversely correlated with the present observation that glomerular density, number and area were greater in the caudal MOB of male than female ferrets. PMID- 15896477 TI - Morphological analysis of the relation between immunoglobulin A production in the small intestine and the enteric nervous system. AB - To investigate the relation between the enteric nervous system and immunocytes, fecal immunoglobulin (Ig) A and the distribution of IgA-positive cells close to nerve fibers in the lamina propria around the crypts of the small intestine were examined after intraperitoneal injection of lipopolysaccharides (LPS). In mice, IgA in the feces 1 h (LPS1) and 6 h (LPS6) after LPS injection increased compared to that in the control group. In the ileum lamina propria of the LPS1 group, the number of IgA-positive cells close to the epithelial basement membrane was increased. In the LPS6 group, on the other hand, there was a significant increase in the number of IgA-positive cells close to both the basement membrane and the nerve fibers. Our data suggest that LPS induced an increase in the number of IgA positive cells migrating to the nerve fibers, and that migration to the nerve fibers is as important as migration to the basement membrane for IgA production. PMID- 15896478 TI - Spinocerebellar ataxia type 2 (SCA2) with white matter involvement. AB - Two sisters presented with olivopontocerebellar atrophy, neuronal loss in the substantia nigra, intranuclear ubiquitin-, ataxin-2-positive inclusions in neurons, and severe demyelination and axon loss of the cerebral white matter with no accompanying inflammatory pathology. The genetic study demonstrated a 22/36 CAG triplet expansion in the SCA2 gene in one of the sisters; SCA1, SCA3, SCA6, SCA7, SCA8, SCA12, SCA17 and DRPL were ruled out in this patient. The present report shows that severe cerebral white matter pathology may occur in the context of SCA2. PMID- 15896479 TI - Characterization of neural cell types expressing peroxiredoxins in mouse brain. AB - The differential expression patterns of antioxidant enzymes observed in the brains of patients with neurodegenerative diseases suggest an important role for reactive oxygen species and antioxidant enzymes in neurodegeneration. The six mammalian peroxiredoxins (Prxs) comprise a novel family of anti-oxidative proteins that are widely distributed in most tissues, but few studies of Prx in brain tissue have been reported. The specific histology of the neural cell types in which Prxs are expressed is an important issue related to biological function and defense against oxidative stress in the brain. This study analyzed mouse brain neural cell types expressing Prx isoforms using single- or double-label immunohistochemical techniques. In neurons, immunoreactivity for Prx II-V was observed in the cytoplasm. In particular, Prx II was found in the habenular nuclei, and Prx III and V were found in the stratum lucidum of the hippocampus. Astrocytes and microglia were immunoreactive only for Prx VI and Prx I, respectively. Prx I and IV immunoreactivity was apparent in oligodendrocytes, where it was principally localized in the nuclei. The observed distribution of Prx isoforms in the mammalian brain may be indicative of their specific roles in their preferred neural cell types and subcellular locales. The results of this study will help in unraveling the physiological and pathophysiological roles of the different Prx isoforms in neural function. PMID- 15896480 TI - Voltage-sensitive dye imaging of intervibrissal fur-evoked activity in the rat somatosensory cortex. AB - The intervibrissal fur-evoked activity in the rat somatosensory cortex was investigated using high-resolution optical imaging with a voltage-sensitive dye. The optical imaging revealed that the intervibrissal fur representation forms a U shaped band around the borders of the posteromedial barrel subfield (PMBSF), and that this representation is characterized by a rostral-to-caudal somatotopic organization. When GABA(A)-mediated inhibition was partially suppressed by treatment with bicuculline, stimulation of the intervibrissal fur elicited spreading of an excitation wave in an area outside the PMBSF. The spreading wave propagated in both directions along the aforementioned U-shaped band of cortex, but barely invaded the center of the PMBSF. These imaging results suggest a distinct subdivision of cortex adjacent to, but outside, the PMBSF in the rat somatosensory cortex; this region receives input from intervibrissal fur, and seems to process its sensory information through well-developed local horizontal connections. PMID- 15896481 TI - Time-efficient localization of the human secondary somatosensory cortex by functional magnetic resonance imaging. AB - Standardized, robust and time-efficient localization of the human secondary somatosensory cortex (S2) is a challenge in clinical blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI). A fully automated tactile stimulation was optimized in seven right-handed volunteers at 1.5 T for minimum scan time, high BOLD signals and robust localization of S2 by systematically varying the applied block-design. All volunteers had six different fMRI measurements of five stimulation-baseline-cycles (sbc) each with equal block duration that was changed between the measurements from 6 s to 30 s. Additional data sets of 4, 3 and 2 cycles were generated post hoc resulting in a total of 168 data sets that were evaluated individually for BOLD-signal intensity (dS%), correlation to the hemodynamic reference function (r) and Euclidean coordinates (x, y, z). Using different block-designs the S2 activation was highly variable regarding the localization rate (lr), the hemispheric symmetry and the BOLD signals. The protocol with 3 cycles, a block duration (dp) of 15 s and a total scan time (dt) of 105 s most robustly localized S2 (contralateral: lr=71.4%, r=0.65, dS=1.01%; ipsilateral: lr=100%, r=0.6, dS=1.14%) whereas the most time efficient protocol to localize SI (sbc=5, dp=6 s, dt=66 s) provided no robust localization of S2. Compared to other published fMRI protocols a scan time reduction up to 86% was achieved. PMID- 15896482 TI - The association between later cortical potentials and later phases of postural reactions evoked by perturbations to upright stance. AB - Previous studies have suggested that early cortical potentials (e.g. N1) that are evoked by perturbations to upright stance are associated with sensory processing of the initial perturbation and that later potentials may represent cognitive processing of this perturbation. However, it has also been suggested that later cortical potentials could reflect sensory and motor processing of later phases of the postural reaction. The current study set out to provide additional insight into the association between perturbation-evoked cortical potentials and postural reactions evoked by whole-body perturbations. By altering the deceleration onset of the perturbation, which altered the timing of later postural responses, we determined whether changes in later postural responses were associated with changes in later potentials. Based on previous work, we hypothesized that later potentials would not be associated with changes in later postural responses. During stance, seven healthy young adults were instructed to maintain their balance following two types of perturbations: (1) acceleration phase immediately followed by a deceleration phase (TASK 1), and (2) acceleration phase followed by a delayed deceleration phase (TASK 2). In spite of profound task differences in later postural responses, results revealed no significant differences in later potentials. This work provides additional support for the idea that latter elements of perturbation-evoked cortical responses are likely independent of evoked motor reactions required to maintain stability. PMID- 15896483 TI - Risk of late-onset Alzheimer's disease associated with BDNF C270T polymorphism. AB - We examined the frequency of the T allele of the C270T polymorphism of the brain derived nerve growth factor (BDNF) gene in a test and replication test design. Our objective was to determine if there is an association between the BDNF gene and Alzheimer's Disease (AD) in a US population. There were 106 autopsy-proven AD cases and 101 controls of similar ages in each test for a total of 212 AD cases and 202 controls. We found that there was a significant increase in the T allele in both the initial set (p=.04) and in the replication set (p=.018). For both groups combined p=.0008. Odds ratio=3.28, 95% CI=1.69-6.34. There were 54 cases of early-onset AD (EOAD) and 159 cases of late-onset AD (LOAD). The results were only significant for LOAD, p=.0002, odds ratio=3.81, 95% CI=1.93-7.52. The r2 or fraction of the variance attributed to the BDNF gene for the LOAD cases was .046. The results were independent of the APOE epsilon4 allele. When the younger controls were removed, providing a close age match to the AD subjects, the frequency of the T allele was even lower and the differences were still significant for both total AD and LOAD cases. In a logistic regression analysis including APOE, age, sex and BDNF, BDNF was significant at p<.0001. We concluded that BDNF gene variants are significant risk factors for late onset AD. PMID- 15896484 TI - Evidence for the involvement of the opioid system in the agmatine antidepressant like effect in the forced swimming test. AB - This study investigated the involvement of the opioid system in the antidepressant-like effect of agmatine in the mouse forced swimming test (FST). The antidepressant-like effects of agmatine (10 mg/kg, i.p.), as well as those of fluoxetine (32 mg/kg, i.p, a selective serotonin reuptake inhibitor, SSRI) or morphine (5 mg/kg, s.c., a nonselective opioid receptor agonist) in the FST was completely blocked by pretreatment of mice with naloxone (1 mg/kg, i.p., a nonselective opioid receptor antagonist). Pretreatment of mice with naltrindole (3 mg/kg, i.p., a selective delta-opioid receptor antagonist), clocinnamox (1 mg/kg, i.p., an irreversible mu-opioid receptor antagonist), but not with 2-(3,4 dichlorophenyl)-N-methyl-N-[(1S)-1-(3-isothiocyanatophenyl)-2-(1 pyrrolidinyl)ethyl]acetamide (DIPPA; 1 mg/kg, i.p., a selective kappa-opioid receptor antagonist) completely blocked the anti-immobility effect of agmatine (10 mg/kg, i.p.) in the FST. These results firstly demonstrate that the antidepressant-like effects of agmatine in the FST seem to be mediated, at least in part, by an interaction with the opioid system, that involves an activation of delta- and mu-opioid receptors. PMID- 15896485 TI - Association study of four activity SNPs of CYP3A4 with the precocious puberty in Chinese girls. AB - CYP3A4 plays an important role in the metabolism of a variety of endogenous and exogenous compounds. Earlier studies revealed that a high-activity variation of the CYP3A4 gene, CYP3A4*1B is significantly associated with the precocious puberty in girls. Other three variations, CYP3A4*4, CYP3A4*5 and CYP3A4*6, which were found in a study carried out in a Chinese population in Taiwan, were reported to down-regulate the enzymatic activity of CYP3A4. The four activity SNPs were typed in our study in two groups of Chinese girls: 176 girls with precocious puberty as the case group, and 192 normal girls as the control group. No variations of CYP3A4*1B and CYP3A4*4 were found in all the cases and controls. Heterozygous of CYP3A4*5 was found in five subjects of the 192 controls but none in the cases, heterozygous of CYP3A4*6 was found in two subjects of the controls but none in the cases. Fisher's exact test showed that the variation of CYP3A4*5 was associated with the onset of puberty in Chinese girls (P-trend=0.038), while the variation of CYP3A4*6 was not associated with the onset of puberty in Chinese girls (P-trend=0.272). The result suggests that these mutations in the CYP3A4 gene have no contribution to the early onset of puberty in Chinese girls, but are related in some way with the puberty development in Chinese girls. PMID- 15896486 TI - Comparative proteomic analysis of the rat spinal cord in inflammatory and neuropathic pain models. AB - Pathological pain associated either with peripheral tissue damage and inflammation (inflammatory pain) or peripheral nerve injury (neuropathic pain) is characterized by persistent pain hypersensitivity. This hypersensitivity is believed to be mediated by sensitization of nociceptors and spinal dorsal horn neurons leading to hyperalgesia and allodynia. Changes of protein expression and/or phosphorylation are known to contribute to the development of this hyperexcitability of the nociceptive system. In the present study we analyzed protein patterns in the spinal cord following paw inflammation or sciatic nerve injury using two-dimensional (2D) gel electrophoresis combined with MALDI-TOF mass spectrometry. 2D-PAGE revealed nine and five regulated proteins following paw inflammation and sciatic nerve damage, respectively. These regulated proteins had not been identified previously with other methods. There was no overlap of regulated proteins between models except for the small heat shock protein alpha crystallin, which was decreased in both models. In conclusion, this study illustrates that employment of the proteomic 2D-PAGE approach allows for identification of novel regulated proteins that may be involved in the central sensitization and possibly manifestation of chronic pain. PMID- 15896487 TI - Skin and platelet alpha-synuclein as peripheral biomarkers of Parkinson's disease. AB - Parkinson's disease (PD) is a heterogeneous disease that can be difficult to diagnose, and for which we have no simple effective biomarker. In this study we have investigated whether peripheral alpha-synuclein might represent a useful biomarker given that it has a central role in the pathogenesis of PD. We found that full length and truncated alpha-synuclein is present in platelets, but the amount is very variable and does not correlate with disease presence or severity. Furthermore, we show that alpha-synuclein can be detected by immunoblotting in some, but not all, human skin biopsies, but again its level does not correlate with disease presence or severity. We conclude that skin or platelet alpha synuclein would not be an appropriate diagnostic biomarker for PD. PMID- 15896488 TI - Differential inhibition of rat alpha3* and alpha7 nicotinic acetylcholine receptors by tetrandrine and closely related bis-benzylisoquinoline derivatives. AB - The patch-clamp technique was used to investigate the effects of bis benzylisoquinoline alkaloids on two of the major neuronal nicotinic acetylcholine receptors (nAChRs), the alpha3-containing nAChR (alpha3*nAChR) endogenously expressed in PC12 cells and the rat alpha7-nAChR heterologously expressed in GH4C1 cells. Tetrandrine and hernandezine reversibly inhibited both receptors displaying half-maximal inhibitory concentrations (IC50) of 8.1 microM and 5.8 microM for alpha3*nAChR and 407.4 nM and 372.2 nM, respectively, for alpha7 nAChR. E6-berbamine completely inhibited the alpha3*nAChR with an IC50 of 5.1 microM, but only partially inhibited the alpha7-nAChR at concentrations up to 30 microM. Tetrandrine inhibition of alpha3*nAChR was functionally non-competitive. All three compounds displaced radiolabelled methyllycaconitine ([3H]-MLA) binding to alpha7-nAChR providing some evidence of competitive antagonism. The results demonstrate that these alkaloids are nAChRs antagonists, with tetrandrine and hernandezine displaying selectivity for one of the major neuronal subtype, the alpha7 nAChR. The different potencies and multiple modes of action on nAChRs may help to better understand the pharmacology of these receptors and to aid in novel drug design. PMID- 15896489 TI - Intrathecal administration of a novel apoE-derived therapeutic peptide improves outcome following perinatal hypoxic-ischemic injury. AB - Perinatal hypoxic-ischemic brain injury remains a significant clinical problem for which there remains no adequate therapeutic intervention. Apolipoprotein E (apoE) is a 299 amino acid protein that has been demonstrated to modify functional recovery following acute ischemic and traumatic brain injury. The aim of the current study was to evaluate whether administration of an apoE-derived peptide could reduce CNS injury in a rodent model of perinatal hypoxia and ischemia. We found that intrathecal delivery of an apoE-mimetic peptide caused a significant reduction in post-ischemic brain necrosis, as reflected by decreased reduction in ipsilateral brain weight 7 days following hypoxic-ischemic injury. These results suggest that administration of an apoE-derived therapeutic peptide represents a novel therapeutic strategy in the clinical setting of perinatal hypoxic-ischemic injury. PMID- 15896490 TI - Intelligence related differences in EEG-bandpower. AB - Several studies on the relationship between event-related desynchronization/synchronization (ERD/ERS) and cognitive performance revealed contradictory results particularly for the alpha band. Studies from our laboratory have shown that good performers show a larger upper alpha ERD (interpreted in terms of larger cortical activation) than bad performers. In contrast, other researchers found evidence for the neural efficiency hypothesis, which states that more intelligent subjects exhibit a smaller extent of cortical activation, which is assumed to be reflected by a smaller upper alpha ERD. Here we address the question whether these divergent results may be due to differences in general task difficulty. Using a modified version of the RAVEN, individually divided into easy and difficult tasks, a group of average and a group of highly intelligent subjects (IQ- and IQ+) have been investigated. While in the theta frequency IQ+ subjects generally exhibited a significantly stronger activation, we found a significant interaction of task difficulty and IQ group in the upper alpha band, indicating both, a weaker activation for the high IQ group during the easy tasks, and a significant increase from easy to difficult tasks for IQ+ only. PMID- 15896491 TI - Comparison of the effects of nicotine and epibatidine given in combination with nomifensine on rotational behaviour in rats. AB - We studied the effects of nicotine and epibatidine on rotational behaviour in 6 hydroxydopamine-lesioned rats. The nicotinic agonists were given acutely and repeatedly either alone or in combination with a dopamine uptake inhibitor, nomifensine. Neither nicotine nor epibatidine alone produced acutely ipsilateral rotational behaviour in rats. However, when given repeatedly once a day for five days, nicotine (0.5 mg/kg) and epibatidine (0.6 microg/kg but not 3.0 microg/kg) induced ipsilateral rotations indicating sensitization of the nigrostriatal dopaminergic system to activation by repeated treatment with nicotinic agonists. Nomifensine (3 mg/kg) alone induced acutely ipsilateral rotations and the effect was enhanced after repeated administration of nomifensine. When the combination of nomifensine and nicotinic agonists was given acutely, the rotational effect of nomifensine was enhanced by nicotine (0.5 mg/kg) but not by either dose of epibatidine. When given repeatedly, nicotine and epibatidine 0.6 microg/kg tended to prolong whereas epibatidine 3.0 microg/kg tended to inhibit the enhanced rotational response to repeated nomifensine. The sensitization of the nigrostriatal dopaminergic pathway might be involved in the beneficial effects of nicotine in Parkinson's disease by allowing the remaining dopaminergic synapses to compensate for the functional deficiency of nigrostriatal pathway. PMID- 15896492 TI - Inhibitory effect of gaseous neuromodulators in vasopressin and oxytocin release induced by endotoxin in rats. AB - Nitric oxide (NO) and carbon monoxide (CO) are endogenously synthesized gaseous molecules that act as neurotransmitters in central nervous system. In this study we investigated the modulatory role of NO and CO in lipopolysaccharide (LPS) induced vasopressin and oxytocin secretion. Intracerebroventricular (i.c.v.) injection of N omega-L-nitro-arginine methyl ester (L-NAME), 3-morpholino sydnonimine (SIN-1), zinc deuteroporphyrin 2,4-bis glicol (ZnDPBG) or hemin did not change the basal vasopressin and oxytocin plasma levels. After endovenous LPS administration, plasma vasopressin and oxytocin increased, reaching a peak at 60 min, and returning to basal levels afterwards. LPS administration induced a higher vasopressin and oxytocin plasma levels in rats previously treated with L NAME and ZnDPBG (P<0.05) compared to rats pre-treated with vehicle. On the other hand, in rats previously treated with SIN-1 or hemin, there was a significant reduction in the vasopressin and oxytocin secretion. These findings confirm the inhibitory role of NO and CO in the LPS-induced vasopressin and oxytocin secretion. PMID- 15896493 TI - Evidence of roles of central alpha1-adrenoceptors and epinephrine in orexin A induced hyperactivity in mice. AB - Previous studies have shown that central alpha1-adrenoceptor activity is necessary, acutely, for gross behavioral activity in response to novel surroundings and various psychostimulants. The present experiment tested whether it is also necessary for the hyperactivity produced by the peptide, orexin A, which is present in several central monoaminergic nuclei. Mice, pretreated intraventricularly with the alpha1-antagonist, terazosin, or the alpha2 antagonist, atipamezole, were given orexin A, intraventricularly (i.v.t.), and videotaped for gross movement and locomotion in the home cage between 30 and 60 min post-infusion. The alpha1-antagonist was found to produce a significant dose dependent decrease of orexin A-induced activity, which was first seen at the 3 nmol dose and was near total at 30 nmol. The alpha2-antagonist, at 10 nmol, had no effect on the orexin A response. Pharmacological inhibition of the synthesis of epinephrine, a potential neurotransmitter at central motoric alpha1 adrenoceptors, with 2,3-dichloro-alpha-methylbenzylamine also significantly attenuated orexin A-induced hyperactivity. It is concluded that central alpha1 adrenoceptor activity, presumably caused by epinephrine release, is necessary for the gross behavioral activation produced by orexin A. PMID- 15896494 TI - Visceral nociceptive input to the area of the medullary lateral reticular nucleus ascends in the lateral spinal cord. AB - In halothane-anesthetized rats, neurons stereotaxically located in the region of the medullary lateral reticular nucleus (LRN) and responsive to urinary bladder distension (UBD) were characterized using extracellular electrodes. Most neurons excited by UBD were also excited by noxious stimuli applied to bilateral receptive fields comprising at least half of the body surface. These bilateral nociceptive specific (bNS) neurons exhibited graded responses to graded intensities of UBD. Neuronal responses to noxious UBD were highly positively correlated with responses to noxious colorectal distension, suggesting a convergence of visceral sensory information in the area of LRN. Bilateral lateral mid-cervical spinal cord lesions virtually abolished activity of bNS neurons evoked by noxious UBD, while dorsal midline lesions had no significant effect. These data support a role for neurons in the region of the LRN in visceral nociception and implicate traditional lateral spinal cord pain pathways in the transmission of visceral information to caudal ventrolateral medullary structures. PMID- 15896495 TI - Effects of endocannabinoids on discharge of baroreceptive NTS neurons. AB - Previously, we have shown that microinjection of endocannabinoids (ECBs) into the nucleus tractus solitarius (NTS) can modulate baroreflex control of blood pressure (BP), prolonging pressor-induced inhibition of renal sympathetic nerve activity. This suggests that ECBs can modulate excitability of baroreceptive neurons in the NTS. Studies by others have shown that neural cannabinoid (CB1) receptors are present on fibers in the NTS, suggesting that some presynaptic modulation of transmitter release could occur in this region which receives direct afferent projections from arterial baroreceptors and cardiac mechanoreceptors. This study, therefore, was performed to determine the effects of ECBs on NTS baroreceptive neuronal discharge. Picoinjection of the ECB anandamide (AEA) was found to significantly increase discharge of baroreceptive neurons (20 of 23). Picoinjection of the ECB uptake inhibitor, AM404, which enhances endogenous ECB activity, also significantly increased discharge of baroreceptive neurons (8 of 10 neurons). To determine if effects of ECBs involved a GABAA mechanism, the neuronal responses to AEA and AM404 were tested after prior blockade of postsynaptic GABAA receptors by bicuculline (BIC) or SR 95531 hydrobromide (gabazine--SR 95531), which would eliminate any effects due to modulation of GABA input. The increase in neuronal discharge to both AEA and AM404 was significantly attenuated following BIC or SR 95531, which alone significantly increased discharge of baroreceptive neurons tested. These results support the hypothesis that ECBs enhance baroreflex function through increases in NTS baroreceptive neuronal activity, due in part to modulation of GABAergic inhibitory effects at the neuronal level. PMID- 15896496 TI - A study of the association of (Val66Met) polymorphism in the brain-derived neurotrophic factor gene with alcohol dependence and extreme violence in Chinese males. AB - From studies of genetic-knockout animals, brain-derived neurotrophic factor (BDNF), a member of the neurotrophin growth-factor family, has been implicated in both alcohol preference and aggressive behaviour. To test whether a BDNF genetic variant may be associated with alcohol-dependent and violent behaviours, we studied Val66Met polymorphism of the BDNF-gene in 110 cases of alcohol dependence, in 134 extremely violent convicts, and in 149 individuals without psychosis or mood disorders. We also examined the association of this polymorphism with antisocial personality disorder comorbidity in the extremely violent convicts. The results showed that the genotype and allele frequencies for Val66Met polymorphism at the BDNF-gene site did not differ among the three groups. Furthermore, it was not demonstrated that this polymorphism is associated with antisocial personality disorder comorbidity in the extremely violent convicts. Based on these findings, it seems reasonable to suggest that this BDNF gene Val66Met polymorphism is unlikely to play a major role in the genetic susceptibility to the traits of alcohol-dependence or violence proneness. PMID- 15896497 TI - Preconditioning-specific reduction of c-fos expression in hippocampal granule and pyramidal but not other forebrain neurons of ischemic brain: a quantitative immunohistochemical study. AB - To specify targets for an ischemic preconditioning paradigm (ischemic tolerance), c-fos expressions in ischemic (induced by 10 min bilateral carotid-occlusions subsequent to coagulation of vertebral arteries) and preconditioned rats (treated for 4 min carotid-occlusions 72 h before ischemia) were compared in 12 forebrain areas/nuclei. Fos immunostaining was applied to serial sections of the forebrain and the density (cell number/area measured) of Fos-immunopositive (Fos+) neurons, as well as their percentile changes were determined in five hippocampal and seven extrahippocampal areas/nuclei of ischemic and preconditioned rats. The ratio of counts found in ischemic over control animals showed several fold increase of Fos+ cells in the three layers (granule cell, molecular and polymorphic) of the dentate gyrus, CA3 and CA1 pyramidal neurons, as well as in thalamic and hypothalamic nuclei and limbic cortical areas. In contrast, preconditioning did not alter c-fos expressions significantly in the extrahippocampal brain areas investigated. These results strengthen the hypothesis that the hippocampal and dentate neurons are more susceptible to ischemic tolerance than cells in other brain regions. In fact stress-response and induction of ischemic tolerance in different forebrain areas can be distinguished. PMID- 15896498 TI - Nicotinic ACh receptors in area postrema neurons of immature rat brain. AB - The electrophysiological properties of nicotinic ACh receptors (nAChR) were investigated in acutely dissociated area postrema (AP) neurons of the immature rat brain using the whole-cell patch-clamp recording method. ACh induced a transient inward current exhibiting a strong inward rectification. The ACh response was mimicked by nicotine and cytisine, and was inhibited by nAChR antagonists, but not by 10(-7) M atropine. Muscarinic AChR agonists did not induce any current. We confirmed the Ca2+ permeability of nAChR. These results indicate the presence of nAChR on AP neurons, and suggest that the activation of nAChR play important roles in cardiovascular functions in rats. PMID- 15896499 TI - The differential JunB responses to inhibition of succinate dehydrogenase in rat hippocampus and liver. AB - The inhibitor of mitochondrial enzyme succinate dehydrogenase, 3-nitropropionic acid (3-NPA), induces cellular energy deficit followed by oxidative stress, secondary excitotoxicity and neuronal degeneration. The fast activation of Jun and Fos proteins and other proteins encoding inducible transcription factors (ITFs) occurs in most tissues upon exposure to a variety of stressors including exposure to mitochondrial inhibitors. However, the consequences of this activation can differ dramatically in different organs. For example, while activation of the same ITFs may lead to apoptosis and necrosis in neurons it may stimulate liver regeneration. Here, we report the alterations in mRNAs levels of c-Fos, JunB, and Krox20 proteins induced in the rat brain and liver by the acute exposure to 3-NPA at 30 mg/kg, s.c. While the increase of c-fos transcripts was observed in both the hippocampus and liver, the junb transcript increased in the hippocampus but decreased in the liver. No changes were observed in krox-20 mRNA in the hippocampus. Interestingly, there was a large variation in krox-20 mRNA levels in the liver among animals within the same experimental group. In conclusion, out of the three ITFs transcripts examined here junb may activate different pathways depending on the tissue as indicated by differential responses to mitochondrial inhibition in the hippocampus and liver. PMID- 15896500 TI - Paediatric burn injuries-tomorrow is too late. PMID- 15896501 TI - Burn scar neoplasms: a literature review and statistical analysis. AB - BACKGROUND: Although squamous cell carcinoma (SCC) is the most common burn scar neoplasm, other neoplasms have also been reported. OBJECTIVE: To compile the burn scar neoplasm cases in the literature and to analyze their frequency and demographic characteristics. MATERIALS AND METHODS: Cases were obtained through literature searches. RESULTS: There were 412 cases gleaned from 146 articles between 1923 and 2004. Seventy-one percent (293) of the tumors were squamous cell carcinoma, 12% (48) were basal cell carcinoma (BCC), 6% (23) were melanoma, 5% (21) were sarcoma, 4% (16) were other neoplasms, 1% (6) were squamo-basal cell carcinoma, and 1% (5) squamous cell-melanoma. The mean age at tumor diagnosis was 50 years, the mean age at the time of burn injury was 20 years, the mean latency interval was 31 years. Only 5% of the reported cases were excised and grafted at the time of injury (p<0.001). BCC occurred at a significantly later age compared to SCC and sarcoma groups (p<0.02) and had a shorter latency period compared to SCC and sarcoma groups (p<0.004) and melanoma (p<0.008). Local recurrence was present in 16% of the cases, while regional lymph node involvement was seen in 22% of the cases and distant metastases was present in 14% of the patients. The mortality rate was 21%. CONCLUSIONS: Although the main burn scar neoplasm was SCC, other neoplasms contributed to the morbidity and mortality of burn patients. Excision and grafting of deep burns, and close follow-up of non-healing ulcerated burn scars is essential for the prevention and early detection of burn scar neoplasms. PMID- 15896502 TI - Prediction of burn healing time using artificial neural networks and reflectance spectrometer. AB - BACKGROUND: Burn depth assessment is important as early excision and grafting is the treatment of choice for deep dermal burn. Inaccurate assessment causes prolonged hospital stay, increased medical expenses and morbidity. Based on reflected burn spectra, we have developed an artificial neural network to predict the burn healing time. PURPOSE: Our study is to develop a non-invasive objective method to predict burn-healing time. METHODS AND MATERIALS: Burns less than 20% TBSA was included. Burn spectra taken on the third postburn day using reflectance spectrometer were analyzed by an artificial neural network system. RESULTS: Forty one spectra were collected. With the newly developed method, the predictive accuracy of burns healed in less than 14 days was 96%, and that in more than 14 days was 75%. CONCLUSIONS: Using reflectance spectrometer, we have developed an artificial neural network to determine the burn healing time with 86% overall predictive accuracy. PMID- 15896503 TI - Is surgery an effective and adequate treatment in advanced Marjolin's ulcer? AB - Malignancies in scars are generally known as Marjolin's ulcers. Between 1999 and 2004, 15 patients with Marjolin's ulcer were treated in our clinic. All lesions were secondary to burns of various causes. We perform a combined approach and aggressive surgery for treatment of Marjolin's ulcer; excision with safe margin, lymphatic dissection, postoperative radiotherapy, chemotherapy and amputation if needed. We think that the scar tissue acts as a barrier for the tumors, which will enlarge. We believe that, if we release this barrier like scar tissue, the virulent the spread of the tumor will be permitted. In this article, we consider whether or not surgical excision alone as recommended in the treatment of Marjolin's ulcers is adequate and effective. An aggressive combined approach is essential for treatment in early stages with high success rate. But there is no consensus for the treatment of advanced disease and results are generally unsuccessful. PMID- 15896504 TI - A global study of hospitalized paediatric burn patients. AB - The global incidence of hospitalized paediatric burn patients is unknown. In order to determine a working estimate of the size of the problem, one approach is to extrapolate from relevant published studies. A literature search of the Medline database was performed to identify epidemiological papers published since 1990, which addressed paediatric burn admissions. Extrapolation from population based studies allowed global figures to be estimated. The highest incidence of hospitalized paediatric burns patients is in Africa, the lowest in the Americas. Europe, the Middle East and Asia share similar figures, but the considerably larger population of Asia indicates that it bears over half of the world's paediatric burn population. It is unfortunate that despite many published studies describing burn admission, few meet criteria that allow for comparative epidemiological data. More attention needs to be focused on uniformity in data collection and presentation to enhance the value of the data available. PMID- 15896505 TI - Management of high voltage electrical injury in children. AB - High voltage electrical injuries in children are rare. They produce severe internal damage to limbs resulting in a high incidence of amputation. Early, aggressive serial debridement ensures adequate removal of dead tissue. Flap coverage to defects is necessary. Secondary surgery is common, and delayed sequelae such as tendon rupture should be expected. PMID- 15896506 TI - Application of tissue ultrasound palpation system (TUPS) in objective scar evaluation. AB - One of the characteristics of hypertrophic scarring is its raised appearance. Its maturation often results in increased thickness. Therapists usually rely on subjective observation and palpation to document scar thickness. The result of these subjective assessments may reflect only the superficial scar thickness but is unable to measure the whole scar volume and thickness under the skin surface. Measurement of scar thickness using ultrasound imaging has been previously reported, but has not been commonly used in clinics due to its complex operation method and high cost. In this study, we have adopted a newly developed and user friendly Tissue ultrasound palpation system (TUPS) for the assessment of scar thickness. It consists of a finger size palpation probe, connected to an ultrasound transducer and an in-series load cell to measure the thickness of the soft tissue over the human body. The method of operation is easy and it can be used to measure skin thickness on various parts of the body, thus reflecting the skin thickness. The reliability of the TUPS in clinical application was tested on 30 subjects with a hypertrophic scar at a local hospital. Three raters implemented two assessments on each subject to study its test-retest and inter rater reliabilities. It was then used to assess 100 subjects with various severity of hypertrophic scar caused by trauma, scald, burn or surgery. They were assessed using TUPS as well as the Vancouver Scar Scale (VSS) for rating scar thickness, pliability, pigmentation and vascularity. Two-way mixed intra-class correlation showed a high test-retest reliability with Intraclass Correlation Coefficient (ICC)=0.98 and inter-rater reliability ICC=0.84. Fair positive correlations with VSS thickness score and VSS total score r=0.34 (p<0.05) and 0.42 (p<0.05), respectively. A significant difference between two scar type groups (50 burn scald scars and 50 surgical scars) was demonstrated (d.f.=52.94, t=3.99, p<0.01). TUPS was proved to have high inter-rater, test-retest reliability and it had a moderate correlation with the VSS that clinicians used for assessment of the scar. This system is recommended for clinical assessment of scar thickness. PMID- 15896507 TI - Ligustrazine attenuates acute lung injury after burn trauma. AB - Acute lung injury is a common complication in patients with extensive burns in which the burned area exceeds 30% of the total body surface area (TBSA). This study was undertaken to evaluate the effect of Ligustrazine on burn-induced lung injury as well as the release of interleukin-8 (IL-8) in rats to characterize the role of Ligustrazine and IL-8 in lung injury after burn trauma. Sprague-dawley rats were divided into three groups: (1) sham group, rats who underwent sham burn; (2) control group, rats given third-degree burns over 30% TBSA and lactated Ringer solution for resuscitation; and (3) Ligustrazine group, rats given burn injury and lactated Ringer's solution with Ligustrazine inside for resuscitation. Pulmonary injury was assessed at 24 h by pulmonary capillary permeability determined with fluorescein isothiocyanate-labeled albumin and lung histologic analysis, and lung myeloperoxidase (MPO) activity as well as lung wet/dry weight ratio. The IL-8 levels were measured in serum by enzyme-linked immunosorbent assay. These studies showed that burn trauma results in increased pulmonary leakage permeability and lung wet/dry ratio, elevated serum IL-8 levels and MPO activity, and worsened histologic condition. Ligustrazine inhibited these changes, prevented burn-mediated lung injury, and the production of IL-8. This will likely provide further evidence for ligustrazine as a therapeutic strategy in burn-induced lung injury. PMID- 15896508 TI - Endothelial thrombomodulin (CD 141) in a rabbit burn model. AB - Thrombomodulin (CD 141) is an endothelial surface transmembrane glycoprotein. It is involved in the activation of protein C in the inactivation of thrombin. In severe sepsis CD 141 is shed from the endothelial surface. This leads to disseminated intravascular coagulation (DIC), disturbed organ functions and multi organ failure (MOF). In this study, we investigated if endothelial bound thrombomodulin is shed in thermal injuries. MATERIAL AND METHODS: In 10 New Zealand white rabbits full thickness and superficial partial thickness burns were produced. Dermal blood flow was analyzed by measuring the fluorescence of intravenously injected indocyanine green. Skin-biopsies were taken from the burn wounds from the zones of stasis between full thickness burns and from unburned skin 72 h post burn. Specimens were processed for immunoperoxidase staining using a specific monoclonal antibody against CD 141. RESULTS: Dermal blood flow was reduced in burned skin areas and in the zones of stasis. Thrombomodulin was only detectable on the surface of capillary endothelial cells in specimens taken from unburned skin areas. No thrombomodulin was detectable in specimens taken from burn wounds or from the zones of stasis. Thus, shedding of thrombomodulin was detectable in areas with reduced dermal blood flow. CONCLUSION: Thermal injuries affect the dermal endothelial surfaces resulting in a shedding of thrombomodulin. This mechanism might be involved in the development of progressive skin damage in the zone of stasis. Disseminated intravascular coagulation following inactivation of thrombomodulin might lead to multiple organ dysfunctions in severe burn injuries. PMID- 15896509 TI - A retrospective analysis of 19,157 burns patients: 18-year experience from Hallym Burn Center in Seoul, Korea. AB - This is a retrospective, epidemiological study of 19,157 acute burn patients admitted to the Hallym Burn Centre, Hangang Sacred Heart Hospital, Seoul, Korea, during the period 1986-2003. The study was conducted to identify characteristics of burn patients that could lead to development of preventive programs to reduce both the frequency and mortality of burn injury. The majority of patients admitted had relatively minor burns, which were treated conservatively. Males predominated in all age groups with a summer peak seasonal variation. Typical burn in the children <5 years of age was a scald injury, occurring at home, affecting upper limbs and resulting in a relatively short hospital stay. Electrical burns due to steel chopsticks and steam burns due to electric rice cookers were also found in this age group. Adult burns, affecting upper and lower limbs, were caused by flames, hot liquids or electricity. Burns were sustained at the workplace and outdoors. Flame accidents in males were the leading cause of death with overall mortality of 8.2%. 13.9% of admissions needed intensive care and average length of hospital stay was 32.6 days. The annual number of patients undergoing burn related surgery has continuously increased reflecting a more aggressive surgical management. Our ongoing efforts are to promote prevention program and look for changes in the incidence of burn injury based on these epidemiologic features. Further study for implications of changes in management protocols and policies on treatment outcomes would improve the provision of appropriate care for the patients. PMID- 15896510 TI - Case-controlled study of patients with self-inflicted burns. AB - The main objectives of this study were to investigate whether patients with self inflicted burns have larger burns, and a worse outcome, than patients with accidental burns. The secondary objective was to examine patient pre-injury characteristics to identify ways of preventing the burn occurring. A case controlled study was performed: 36 deliberate self-burn patients were matched separately to two groups of accidental burn patients. The first group was used to compare burn severity. Patients were matched for age and sex; they were excluded if they had a psychiatric diagnosis, or a non-burn injury. The second group was used to compare outcome. The same matching and exclusion criteria were used as in the first group, with the addition of burn-size. Deliberate self-burn patients have significantly larger burns (p<0.01; median total body surface area (TBSA) 10% versus 1.5%) than accidental burn patients. They also stay in hospital longer, even when matched for burn-size (p<0.02; median stay 15 days versus 9 days). Self-inflicted burns occurred in supervised environments in 28% of cases. The number of deliberate self-burns could be reduced with simple interventions such as restricting smoking in hospitals and prisons, and also by identifying high-risk patients. The poor outcome from deliberate self-burns could be improved by well-coordinated multidisciplinary patient management with early psychiatric team involvement. PMID- 15896511 TI - Microdialysis for detection of dynamic changes in tissue histamine levels in experimental thermal injury. AB - Histamine is an important mediator contributing to oedema formation after thermal injury. Tissue histamine concentrations have been previously determined by analyzing tissue biopsies. The microdialysis method enables continuous collection of samples from the extracellular tissue fluid. In this experimental burn study on pigs samples from the extracellular fluid for histamine analysis were collected from superficial, partial thickness and full thickness burn sites during a 24-h period. There was a burn depth-related increase in histamine concentrations during the first 2 h post injury. Deep burns induced a more profound initial increase in tissue histamine concentration than the partial thickness and superficial burns. Histamine concentrations at all burn sites declined until 12 h post injury. There was a second rise in tissue histamine concentrations between 12 and 24 h post injury without a rise in plasma histamine concentrations. Histamine concentrations at all burn sites were higher than at the non-burned control sites. The microdialysis technique is an easily applicable method of collecting on-line samples from burned tissue. This method provides a useful tool in investigating the effects of different treatment modalities on the secretion of substances into interstitial fluid within burned tissue. PMID- 15896512 TI - Prebiotic ingestion does not improve gastrointestinal barrier function in burn patients. AB - Prebiotics increase intestinal levels of health-promoting bacteria implicated in decreasing pathogen colonization, stimulating immune functions and stabilizing gut barrier functions, parameters which are altered in burn patients. We propose that regular intake of a prebiotic, oligofructose (OF), might help to improve the altered gastrointestinal (GI) permeability observed in burn patients. A randomized, double-blind, controlled clinical trial was carried out in 41 burn patients (mean burn surface area=17.1+/-8.2%) who ingested daily 6 g of oligofructose (OF group) or sucrose as placebo (Control group) during 15 days. Gastrointestinal permeability to sucrose and lactulose/mannitol (L/M) was evaluated on days 1 (before treatment) 3, 7, 14 and 21. A permeability test was also performed in 18 healthy subjects as controls. Thirty-one patients completed the protocol (dropout rate=24.4%). Healthy subjects had a basal sucrose excretion of 21.3 mg (14.0-32.5 mg) and a basal L/M ratio of 0.017% (0.009-0.022%). Sucrose excretion increased 5-fold and L/M ratio 4.4-fold in burn patients on day 1 and these high levels of marker excretion decreased significantly throughout the study (p=0.016 and 0.000001, respectively). No differences between the OF and Control groups were observed for sucrose excretion or L/M ratio. In conclusion, the normalization of gastrointestinal permeability is not accelerated by prebiotic intake. PMID- 15896513 TI - Povidone-iodine ointment: no effect of split skin graft healing time. AB - In major burns, local treatment of the split skin graft after a burn injury is important to prevent serious infectious complications. Topical burn wound therapy may improve the bacteriological condition of the wound, which in turn may improve the successful take of a skin graft. Delay of wound healing is an undesired side effect of these topical anti-microbial agents. It is known that povidone-iodine has many clinical advantages. In view of this, the total healing time of the freshly grafted burn wound was studied. In this prospective study, comparable areas of the same patient were treated with povidone-iodine ointment or with simple vaseline gauze. There was no statistical difference in the total wound healing time between the treated and the control group. PMID- 15896514 TI - Eyebrow reconstruction following burn injury. AB - Facial burns represent between one-fourth and one-third of all burns. Absence of the eyebrows or distortions in their position alter the character of the face. Thus, eyebrow repair or reconstruction can be an important "finishing touch" in the overall reconstruction of a burned face. Generally, there are three ways to reconstruct the eyebrow: use of superficial temporal artery island flap; composite graft from scalp; and mini or micrografts from scalp. This report presents 20 patients reconstructed with the above techniques. Eleven patients (eight male and three female) with superficial temporal artery island flaps; seven patients (two male and five female) with composite grafts from scalp; and two patients (female) with minigrafts. The results suggest that superficial temporal artery island flaps were more suitable for males and composite graft for females who generally require thinner and less dense eyebrows. Our experience with minigrafts for burn alopecia has not been adequate. Here in this article, different methods of eyebrow reconstruction are presented with the greater emphasis on superficial temporal artery flap. PMID- 15896515 TI - Re-emphasising the efficacy of the multi-purpose, self-adjustable, aeroplane splint for the splinting of axillary burns. AB - This article re-emphasises the efficacy of the previously described multi purpose, self-adjustable, aeroplane splint for the splinting of axillary burns. This superior to the commonly used contoured splint for various reasons namely (i) its usability during the acute burn care rehabilitation, (ii) its cost effectiveness, (iii) its durability, and (iv) its strength. Above all due to its easy maneuverability this splint provides easy accessibility around environmental barriers. Though we acknowledge that there is a possibility of 'chord of circle' development due to the use of this splint, we have been successful in preventing such a complication simply by prescribing pressure garment concomitantly with this splint. Thus this article highlights the efficacy of earlier described multi purpose, self-adjustable, aeroplane splint for the splinting of axillary burns. PMID- 15896516 TI - Adjustable aesthetic aeroplane splint for axillary burn contractures. AB - Managing axillary burns with an Aeroplane Splint has been known for its effectiveness for years. However, poor compliance in using the various models of currently available aeroplane splints leads to an inadequate outcome, because of the discomfort produced not only in wearing but also while ambulating within the community. In developing countries like India the biggest barrier to environmental accessibility is the presence of multiple permanent structures like narrow thoroughfares, entry/exit sites of public transport, and narrow aisles. These pose a challenge to rehabilitation professionals while prescribing large splints. An attempt to target these problems was made by Manigandan et al, but aeroplane splints, even the one described by them does not deal with the major problem of aesthetic appeal as identified by many of our patients. Accordingly this article presents a new model of the aeroplane splint with innovative changes, focussing on aesthetic appeal while maintaining all the benefits of the splint described by Manigandan et al. PMID- 15896517 TI - A modified technique of percutaneous subclavian venous catheterization in the oedematous burned patient. AB - Infraclavicular subclavian venepuncture in the oedematous burned patient is often difficult because of increased depth of the vein. In addition, proper patient positioning is not easily achieved because of extensive burns, generalised oedema and bulky dressings. To overcome these difficulties, a modified technique of infraclavicular subclavian venepuncture has been developed. The introducer needle is bent to create a mild curvature. It is inserted at a point 1-2 cm inferior to the palpable lower border of the clavicle along the junction of the middle and medial thirds of the bone, advanced along the deep surface of the clavicle and directed at the superior border of the suprasternal notch. This medial point of insertion shortens the distance of access to the subclavian vein. The curve allows the tip to be kept close to the undersurface of the clavicle as the needle is advanced, thereby reducing the risk of injury to deep structures. The advantages of the modified technique are demonstrated in anatomical dissections. This technique is a viable alternative when conventional techniques fail. PMID- 15896518 TI - Reconstruction of nose and lips with tubed cervical flap in electrical injury: a case report. PMID- 15896519 TI - Combined treatment using artificial dermis and basic fibroblast growth factor (bFGF) for intractable fingertip ulcers caused by atypical burn injuries. PMID- 15896520 TI - Burn injury caused by flammable flying toy balloons. PMID- 15896521 TI - Burn injuries caused by fire breathing. PMID- 15896522 TI - Salvage of the severely burned leg with anterolateral thigh flaps. PMID- 15896523 TI - A possible chemical burn to the scalp following hair highlights. PMID- 15896524 TI - The management of an autistic burns patient. A case for ventilation? PMID- 15896525 TI - The Asia pacific burns association. PMID- 15896526 TI - Re-emphasising the use of the cost-effective ear-cup as a pressure-relieving splint in the management of ear reconstruction. PMID- 15896527 TI - Establishing equivalence classes in preschool children with one-to-many and many to-one training protocols. AB - The present study set out to assess if the different probabilities reported in the literature of obtaining equivalence after baseline training with MTO and OTM protocols could be attributed to individual differences and, if so, whether equivalence formation could be facilitated by using familiar stimuli as nodes. In Experiment 1, 16 preschool children were trained on four sets of 2-choice match to-sample tasks, eight with a OTM protocol (A-B, A-C, A-D, A-E) and eight with a MTO protocol (B-A, C-A, D-A, E-A). For four OTM and four MTO children only abstract stimuli were used. The other four OTM children and four MTO children received the same training but with familiar stimuli as nodes. All children received tests for equivalence (first) and symmetry (second). In Experiment 2, eight children who served in Experiment 1 participated again, four who had passed the equivalence test, and four who had failed that test. All children received the same baseline training as in Experiment 1 but with the opposite type of nodes (abstract instead of familiar, and vice versa) and training protocol (MTO instead of OTM, and vice versa). The results showed that (a) the children's performances (training and testing) were not affected by the training protocol; (b) equivalence formation occurred more readily when being trained with all abstract stimuli than when familiar stimuli served as nodes; and (c) most children who passed or did not pass the equivalence test in Experiment 1 repeated their performance in Experiment 2, irrespective of the conditions that were used. PMID- 15896528 TI - A note on select- and reject-controlling relations in the simple discrimination of capuchin monkeys (Cebus apella). AB - Controlling relations in the simple discrimination performances of capuchin monkeys (Cebus apella) were studied in two experiments using a blank-comparison procedure. The main goal was to determine whether monkeys would (a) select an S+ stimulus if another stimulus was substituted for the S- (indicating a select controlling relation) and (b) reject an S- if another stimulus was substituted for S+ (indicating a reject-controlling relation). In experiment 1, two simple simultaneous discriminations were established, one of which was reversed repeatedly until rapid reversal learning was exhibited. During subsequent probe tests, some behavior was consistent with select- and reject-controlling relations, but there was also substantial variability. To control the variability, the procedures of experiment 2 were designed to establish select- and reject-control relations directly by training with the blank-comparison procedure. On subsequent probe trials, new stimuli were substituted for the blank comparison. Both animals exhibited consistent, reliable select- and reject controlling relations. These experiments are the first to employ the blank comparison procedure with non-human subjects. They also demonstrate a reliable method for generating select- and reject-controlling relations for experimental study. PMID- 15896529 TI - The effect of terpenoid extracts from 15 pine species on the feeding behavioural sequence of the late instars of the pine processionary caterpillar Thaumetopoea pityocampa. AB - The feeding behaviour of the pine processionary (PPC) caterpillar Thaumetopoea pityocampa Den. and Schiff. (Lepidoptera, Thaumetopoeidae) in L3-L4 stages was explored by means of laboratory arena feeding trials and natural substrates. In the bioassays, volatile extracts of 15 pine species, 8 of which are naturally growing in Greece, were incorporated. An analytical model was developed based on the principle of multinomial logit regression with five outcomes on the basis of the behavioural feeding sequence of the caterpillars. The outcomes were the five steps in which the feeding behavioural sequence was decomposed. The model's suitability (MacFadden's rho(2)=0.229, P<10(-4)) was examined when including 10 terpenes that were judged significant through a stepwise canonical discriminant analysis. The proposed model was superior to a random one and the two models resulting from the addition and subtraction of 4 terpenes to the already 10 existing compounds. The most informative model was built on the terpenes caryophyllene oxide, terpinolene, myrcene, germacrene D, eudesmol, limonene, beta pinene, beta-caryophyllene, alpha-pinene and manoyl oxide. The background terpenes were present in the model and of particular importance. No special behavioural role, either as promoter or inhibitor could be assigned to the individual volatile metabolites, since no constant pattern among behavioural steps was observed. For instance, beta-caryophyllene while acts as promoter of attraction and trial bite it is a suppressor of partial feeding and strongly inhibits complete needle consumption. The monoterpene limonene, on the other hand, seems to be a suppressor of partial and complete feeding. The overall methodological scheme and the analytical modelling tool could be proved a suitable research protocol in unfolding the ecological role of a complex mixture of secondary metabolites. Those who develop safe practical systems can use this. PMID- 15896530 TI - Mental rehearsal in great apes (Pan troglodytes and Pongo pygmaeus) and children. AB - The ability to rehearse possible future courses of action in the mind is an important feature of advanced social cognition in humans, and the "social brain" hypothesis implies that it might also be a feature of primate social cognition. We tested two chimpanzees, six orangutans and 63 children aged 3-7 years on a set of four puzzle boxes, half of which were presented with an opportunity to observe the box before being allowed to open it ("prior view"), the others being given without an opportunity to examine the boxes before handling them ("no prior view"). When learning effects are partialled out, puzzle boxes in the "prior view" condition were opened significantly faster than boxes given in the "no prior view" condition by the children, but not by either of the great apes. The three species differ significantly in the speed with which they opened boxes in the "no prior view" condition. The three species' performance on this task was a function of relative frontal lobe volume, suggesting that it may be possible to identify quantitative neuropsychological differences between species. PMID- 15896531 TI - The effect of auditory distractors on song discrimination in male canaries (Serinus canaria). AB - Male songbirds such as canaries produce complex learned vocalizations that are used in the context of mate attraction and territory defense. Successful mate attraction or territorial defense requires that a bird be able to recognize individuals based on their vocal performance and identify these songs in a noisy background. In order to learn more about how birds are able to solve this problem, we investigated, with a two-alternative choice procedure, the ability of adult male canaries to discriminate between conspecific song segments from two different birds and to maintain this discrimination when conspecific songs are superimposed with a variety of distractors. The results indicate that male canaries have the ability to discriminate, with a high level of accuracy song segments produced by two different conspecific birds. Song discrimination was partially maintained when the stimuli were masked by auditory distractors, but the accuracy of the discrimination progressively declined as a function of the number of masking distractors. The type of distractor used in the experiments (other conspecific songs or different types of artificial white noise) did not markedly affect the rate of deterioration of the song discrimination. These data indicate that adult male canaries have the perceptual abilities to discriminate and selectively attend to one ongoing sound that occurs simultaneously with one or more other sounds. The administration of a noradrenergic neurotoxin did not impair markedly the discrimination learning abilities although the number of subjects tested was too small to allow any firm conclusion. In these conditions, however, the noradrenergic lesion significantly increased the number failures to respond in the discrimination learning task suggesting a role, in canaries, of the noradrenergic system in some attentional processes underlying song learning and processing. PMID- 15896532 TI - Response-cost punishment via token loss with pigeons. AB - Two experiments were conducted to investigate punishment via response-contingent removal of conditioned token reinforcers (response cost) with pigeons. In Experiment 1, key pecking was maintained on a two-component multiple second-order schedule of token delivery, with light emitting diodes (LEDs) serving as token reinforcers. In both components, responding produced tokens according to a random interval 20-s schedule and exchange periods according to a variable-ratio schedule. During exchange periods, each token was exchangeable for 2.5-s access to grain. In one component, responses were conjointly punished according to fixed ratio schedules of token removal. Response rates in this punishment component decreased to low levels while response rates in the alternate (no-punishment) component were unaffected. Responding was eliminated when it produced neither tokens nor exchange periods (Extinction), but was maintained at moderate levels when it produced tokens in the signaled absence of food reinforcement, suggesting that tokens served as effective conditioned reinforcers. In Experiment 2, the effect of the response-cost punishment contingency was separated from changes in the density of food reinforcement. This was accomplished by yoking either the number of food deliveries per component (Yoked Food) or the temporal placement of all stimulus events (tokens, exchanges, food deliveries) (Yoked Complete), from the punishment to the no-punishment component. Response rates decreased in both components, but decreased more rapidly and were generally maintained at lower levels in the punishment component than in the yoked component. In showing that the response-cost contingency had a suppressive effect on responding in addition to that produced by reductions in reinforcement density, the present results suggest that response-cost punishment shares important features with other forms of punishment. PMID- 15896533 TI - Foot-use laterality in the Japanese jungle crow (Corvus macrorhynchos). AB - Laterality of foot-use behaviour was examined in jungle crows (Corvus macrorhynchos). Consistency of laterality was also verified in comparison with two different behaviours. One was beak-scratching, and another was food-holding. Significant bias of foot-use laterality was found in 11 of 13 tested subjects for beak-scratching and in 10 of 12 tested subjects for food-holding. Bias of foot use laterality was not common among footed subjects, suggesting no population level bias. However, a significant number of footed subjects (10 of 12) who were tested in both behaviours showed consistent laterality irrespective of behavioural differences. These results suggest that jungle crows have strong individual-level laterality in foot-use behaviour. PMID- 15896534 TI - Impaling in true shrikes (Laniidae): a behavioral and ontogenetic perspective. AB - The impaling of prey is a behavioral trait restricted to the true shrikes (Laniidae). Here, we suggest the ontogeny of this behavior. We believe impaling originated from wedging behavior that occurs among several other groups of birds, including corvids. Accidental impaling during wedging was likely the behavioral precursor of purposeful impaling. Fidelity to impaling sites lead to the creation of caches, which were eventually used by females for male evaluation. Caching allowed males to increase their fitness by using the caches as a display to attract potential mates. Further, caching is used by shrikes to demarcate territories, store food for inclement weather or periods of stress in the breeding cycle, divide labor between the breeding pair, and for "aging" while deterrent chemicals in prey decompose. PMID- 15896535 TI - Summation of latent inhibition and overshadowing in a generalized bait shyness paradigm of rats. AB - This paper reports a single experiment using four groups of rats. A group of thirsty rats showed aversion to sodium chloride (NaCl) solution in testing after drinking lithium chloride (LiCl) solution, demonstrating generalized bait shyness due to the common "salty" taste. The other two groups of rats were treated identically except that the rats were either exposed to NaCl solution prior to drinking the LiCl solution or allowed to drink LiCl-sucrose solution instead of the LiCl solution. The weaker salt aversion observed in these groups indicates latent inhibition and overshadowing, respectively. The rats in the fourth group were preexposed to NaCl and allowed to drink the LiCl-sucrose solution. The weakest salt aversion in this group suggests summation of latent inhibition and overshadowing effects. PMID- 15896536 TI - Life-threatening bullous dermatoses: Pemphigus vulgaris. AB - Pemphigus vulgaris (PV) is a rare autoimmune bullous dermatosis with a high mortality rate if untreated. The disease results from autoimmunity to normal components of keratinocyte cell membrane (desmogleins 3 and 1) belonging to the cadherin supergene family. Standard therapy for PV is based on a combined administration of high-dosed glucocorticoids and immunosuppressive drugs. In patients with severe, life-threatening, or recalcitrant PV, stronger therapeutic options should be considered, such as 'pulse-therapy' with discontinuous intravenous infusion of megadoses of immunosuppressive drugs over a short-time, plasmapheresis, and extracorporeal immunoadsorption of pathogenic autoantibodies using the extracellular domain of the PV main antigen (desmoglein 3) produced by baculovirus or, more recently, a tryptophan-linked polyvinyl alcohol adsorber. PMID- 15896537 TI - Life-threatening purpura and vasculitis. AB - Palpable purpura, the inflammation of blood vessels is the hallmark of vasculitis. It can be observed in a variety of settings, where vessels can be affected primarily or as a secondary event. Every patient with vasculitis should be considered to have a systemic disease unless proven otherwise. One or more systemic symptoms occur in at least 50% of patients and there is no way to predict systemic involvement. Patients may demonstrate mild systemic involvement like arthralgia and arthritis, fever and malaise or more severe symptoms such as massive proteinuria and raised creatinine leading to chronic renal failure, severe intestinal bleeding or perforation with a fatal outcome. In this article we will review the life-threatening aspects of purpura and vasculitis. PMID- 15896538 TI - Life-threatening dermatoses due to connective tissue disorders. AB - Connective tissue disorders such as scleroderma, dermatomyositis and lupus erythematosus are autoimmune, multi-system disorders whose clinical manifestations can be restricted to the skin or may involve many organs. The degree and rate of organ system involvement defer, as does the prognosis and rapidity of disease progression. In this article, scleroderma, dermatomyositis and lupus erythematosus will be reviewed in respect to their life-threatening potential. PMID- 15896539 TI - Life-threatening dermatoses in travelers. AB - Life-threatening dermatoses in travelers relates to infections, or allergic or drug reactions. Of the infectious dermatoses, most are parasitic or viral- because of the short time frame. Bacterial infections usually have a longer incubation. More significant are allergic reactions from environmental or dietary causes, and drug reactions, especially those drugs taken for prophylaxis from tropical diseases. All of these can be fulminant and therefore an awareness and early therapeutic intervention is mandatory. PMID- 15896540 TI - Life-threatening mycobacteria infection. AB - Most mycobacteria cause localized and often harmless infections of the skin. Leprosy, which dates back to approximately 60 bc in India, was supposed to be eliminated as a public health problem by the year 2000. With a new case detection rate between 600,000 and 700,000 yearly however, leprosy, with its sometimes devastating consequences, will be with us for many years to come. Buruli ulcer, named after the area in Uganda where prevalence was high, has spread to new areas, especially in Africa. PMID- 15896541 TI - Life-threatening dermatoses due to metabolic and endocrine disorders. AB - Cardiovascular heart disease due to atherosclerosis is the commonest cause of mortality and morbidity in the Western world. Atherosclerosis is directly related to disorders of lipid metabolism, diabetes mellitus (DM), and insulin resistance. Disorders of thyroid function and porphyrin metabolism occur less frequently but may cause life-threatening situations if unrecognized. All these disorders are associated with characteristic dermatoses, which should permit early diagnosis and meaningful intervention. PMID- 15896542 TI - Life-threatening disorders of mucous membranes. AB - Oral mucosa is one of the first barriers to the outside world which encounters various antigens, microorganisms and physical agents. Numerous oral pathologies challenge the dermatologists. Some may be the first sign of an underlying immunosuppression, while others are the inevitable serious outcomes of long lasting mucosal disease. The differential diagnosis is crucial in terms of prompt and effective treatment. PMID- 15896543 TI - Life-threatening environmental dermatoses and agents of mass destruction with serious systemic consequences. AB - The idea of this paper is to acquire a new view of some environmental conditions affecting the skin and carry with them significant morbidity. It is also to understand their etiology and nature to prevent their hazardous and serious complications which are life-threatening. Although most dermatologic disorders rarely have a fatal outcome, there are many potentially fatal medical disorders that have early or major dermatologic manifestations. Some cases lead to death because of the loss of barrier function of the epidermis with fluid and electrolyte loss or from systemic invasion of microorganisms. In others, dermatologic signs reveal an underlying systemic disease where death may occur from the underlying problem. PMID- 15896544 TI - Life-threatening graft-vs-host disease. AB - Hematopoietic stem cell transplant (SCT) is considered standard therapy for a variety of malignant and nonmalignant diseases. Graft-versus-host disease (GVHD) still represents today a major complication of hematopoietic SCT. Two types of GVHD have traditionally been recognized on the basis of the time of onset following transplantation, distinct pathobiological pathways, and different clinical presentations. The acute form commonly breaks out 2 to 6 weeks after transplantation, affecting up to 60% of patients receiving allogeneic transplants from HLA identical donors. Transfer of immunocompetent donor T cells contained in the graft may undergo alloreactivity against recipient cells because of major or minor histocompatibility antigens disparities between the donor and the immunosuppressed host. Target specificity in acute GVHD involves preferential injury to epithelial surfaces of the skin and mucous membranes, biliary ducts of the liver, and crypts of the intestinal tract. Chronic GVHD affects approximately 30% to 80% of patients surviving 6 months or longer after stem cell transplantation and is the leading cause of nonrelapse deaths occurring more than 2 years after transplantation. Chronic GVHD is a multiorgan syndrome with clinical features suggesting some autoimmune diseases, and possibly both alloreactive and autoreactive T cell clones are involved in its pathophysiology. Although GVHD may convey beneficial graft-versus-leukemia/lymphoma effects, it also entails a significant risk of morbidity and mortality. Patients with mild GVHD need only minimal, if any, immunosuppressive treatment, whereas prognosis of patients with extensive disease or resistant to standard immunosuppressive treatment may be dismal. Early recognition of GVHD followed by prompt therapeutic intervention may prevent the progression to higher-grade disease and improve the outcome for patients receiving hematopoietic SCT. PMID- 15896545 TI - Life-threatening paraneoplastic cutaneous syndromes. AB - Paraneoplastic syndromes are diseases or symptom complexes associated with malignancy, usually internal. In dermatology, we modify the definition to refer to dermatoses associated with internal malignancy. In this article, we discuss the link between malignancy and such dermatologic disorders as acanthosis nigricans, acrokeratosis paraneoplastica of Bazex, dermatomyositis, erythema gyratum repens, necrolytic migratory erythema (glucagonoma syndrome), and paraneoplastic pemphigus and discuss, where such information is known, the mechanism by which these paraneoplastic diseases occur. PMID- 15896546 TI - Skin torture. AB - The inhumanity of human beings to other human beings has transcended all continents and every civilization from time immemorial. No amount of prohibitions, be they religious or legal edicts, has prevented cruel and inhuman punishment inflicted by human beings on others to subjugate them. Torture continues to be a common method to subdue and intimidate people. The forms of bodily harm can take any number of ways, some of which might even defy the imagination of any civilized people. We need only to recall the Holocaust and the decimations of thousands in Africa in more recent times. PMID- 15896547 TI - Drug rash with eosinophilia and systemic symptoms vs toxic epidermal necrolysis: the dilemma of classification. AB - According to contemporary vernacular, when the cutaneous manifestations of drug rash with eosinophilia and systemic signs (DRESS) syndrome are those of Stevens Johnson syndrome (SJS) or toxic epidermal necrolysis (TEN), the condition is defined as "DRESS syndrome with severe cutaneous reactions". In this article, we have presented arguments for and against including patients with skin lesions of the SJS/TEN syndromes who also have fever (practically all of the patients) and internal organ involvement (most of the patients) under the definition of DRESS syndrome. After weighing the arguments for and against this alteration of definition, we conclude that it makes more sense for patients with SJS/TEN to be classified as such and not be lumped together under the misleading label of DRESS syndrome. PMID- 15896548 TI - It's time for a change. PMID- 15896549 TI - Purified human plasma glycosaminoglycans limit oxidative injury induced by iron plus ascorbate in skin fibroblast cultures. AB - A number of studies in vivo and in vitro showed that high levels of glycosaminoglycans (GAGs) are found as a consequence of free radical damage. The GAG over production may represent an endogenous mechanism capable to limit oxidative damage. Based on these hypotheses, the aim of this study was to evaluate the antioxidant property of GAGs of human origin in fibroblast cultures. Purified human plasma GAGs were added to the fibroblast cultures in which oxidative stress was induced by the oxidizing system employing iron (Fe2+) plus ascorbate. We assessed cell death, lactate dehydrogenase activity, membrane lipid peroxidation, DNA damage, protein oxidation, hydroxyl radical (OH*) generation and endogenous antioxidant depletion. The exposure of fibroblasts to FeSO4 produced cell death and increased OH* production. It also caused DNA strand breaks and protein oxidation as shown by the DNA fragment analysis and protein carbonyl content, respectively. In addition, FeSO4 enhanced lactate dehydrogenase activity and lipid peroxidation while decreased antioxidant defences. Purified human GAGs, at three different doses, reduced cell death, limited DNA fragmentation and protein oxidation, decreased OH* generation and lactate dehydrogenase activity, inhibited lipid peroxidation and improved endogenous antioxidant defences. These results further support the hypothesis that these molecules may function as antioxidants. PMID- 15896550 TI - The effects of some Mannich bases on heat shock proteins HSC70 and GRP75, and thioredoxin and glutaredoxin levels in Jurkat cells. AB - Mannich bases interact with cellular thiols and inactivate thioredoxin reductase. In this study, the effects of cytotoxic mono-Mannich bases 2, 3 and cyclic Mannich base C1 on the expression of cytoprotective heat shock proteins (HSC70 and GRP75) and on levels of thioredoxin (TRX) and glutaredoxin (GRX) were investigated in Jurkat cells. Cells were exposed to the compounds for 24 h in cell culture medium with 1% FBS. C1 and 2 increased the levels of HSC70 (200% of control) in all the concentrations tested, but 3 did not affect HSC70 levels. Whereas 3 increased GRP75 expression (123-154%), 2 and C1 either did not affect (95-87% for 2, and 88% for C1) or slightly decreased GRP75 expression (82% for 2 and 67% for C1). Mannich bases generally decreased GRX levels (68%, 63-77% and 33 71% for 2, 3 and C1, respectively), but 3 increased GRX levels at 1 microg/ml (142%). Whereas 2 and 3 decreased TRX levels (30-79% and 37-44% of control, respectively), C1 increased the expression of TRX (156-201%). Our results suggest that decreases in GRX and TRX due to the alkylating effects of Mannich bases might have prevented cell division and decreased survival in Jurkat cells, which could not be prevented by increased heat shock protein expression. PMID- 15896551 TI - Styrene and ethylene glycol have a synergetic effect on lipid peroxidation that is better protected than repaired by CoQ10. AB - Previous study of a group of 22 workers occupationally exposed to styrene, ethylene glycol and their mixture at a paint and lacquer industry indicated significantly elevated concentration of malondialdehyde with 4-hydroxynonenal (MDA+4-HNE) in the blood plasma, successfully decreased with coenzyme Q10 (CoQ10) supplementation. The aim of present study is to evaluate whether the exposure to styrene or/and ethylene glycol could be responsible for the increase in MDA level. The mechanism of a single solvent action and the mixture was examined, specially whether it is connected with hydroxyl radical (*OH) generation. It was also investigated whether coenzyme Q10 could be considered as a protective (given before the solvents) or repairing (given after the solvents) agent in oxidative stress caused by the solvents. The results indicate that ethylene glycol nor styrene increase MDA and *OH, but as a mixture give synergetic interaction, elevating MDA and *OH concentration to a statistically significant extent. Coenzyme Q10 at a dose of 3.0 microg/ml only protects, but does not repair increased lipid peroxidation caused by ethylene glycol with styrene. In order to obtain both a protective and repairing effect, a concentration of 12.0 microg/ml CoQ is needed. PMID- 15896552 TI - The effects of ruthenium red, dantrolene and nimodipine, alone or in combination, in NMDA induced neurotoxicity of cerebellar granular cell culture of rats. AB - In the present study, we used the L-type calcium channel blocker nimodipine, the endoplasmic reticulum (ER) calcium release blocker dantrolene and the calcium transport blocker in mitochondria ruthenium red (RuR), in NMDA induced neurotoxicity, to observe the most suitable combination for neuroprotection in cerebellar granular cell culture of rat pups. In addition we tested the effect of RuR on intact neuronal cells without adding NMDA into the flask, in order to compare the effects. As was expected, NMDA induced neuronal cell death. In NMDA induced neurotoxicity, RuR was the most neuroprotective agent of all three compounds tested, but interestingly, RuR alone was found to be neurotoxic in non NMDA treated cultures. RuR showed neuroprotectivity in a dose dependent manner in NMDA toxicity. Dantrolene and nimodipine were also found to have neuroprotective properties in NMDA induced cell death in rat cerebellum. On the other hand, the combined application of the compounds was not found as protective as ruthenium red applications alone. The present study revealed a neurotoxic feature of ruthenium red, however we also demonstrated a neuroprotective role for it in NMDA mediated neuron culture. This could be interpreted as a result of the partial agonistic effect of ruthenium red. PMID- 15896553 TI - Effect of fatty acids on herbicide transport across Caco-2 cell monolayers. AB - Oral ingestion of pesticides can be a major exposure route. These compounds are frequently consumed in the presence of triacylglycerides, which are then hydrolyzed to free fatty acids. The purpose of this work was to examine the effect of two common fatty acids, palmitic (PA) and oleic (OA) acids, and the biological emulsifier sodium taurocholate (TC) on the absorption of three herbicides (trifluralin, alachlor and atrazine) by Caco-2 cell monolayers. Trifluralin's absorption was enhanced (p < 0.05) in the presence of OA whereas the greatest absorption of atrazine and alachlor occurred with PA and the control media, respectively. Trifluralin had significantly lower absorption through the monolayer than either alachlor or atrazine (p < 0.001). A mass balance study demonstrated that trifluralin accumulated within the cell monolayer (13.85% of the donor after 3 h of exposure), but alachlor and atrazine (1.27% and 0.85%, respectively) did not. This response was linear with time (21.89% trifluralin after 6 h of exposure), and demonstrated the potential for continued release of trifluralin after source removal. These experiments demonstrated that fatty acids and an emulsifier can influence absorption of herbicides across small intestinal epithelium. PMID- 15896554 TI - Protective effects of antioxidant vitamins on Aroclor 1254-induced toxicity in cultured chicken embryo hepatocytes. AB - Primary culture of chicken embryo hepatocytes (CEHs) was established to reveal toxicity of polychlorinated biphenyls (PCBs) and attenuating effects of antioxidants vitamin E (VE), vitamin C (VC) and vitamin A (VA) on PCBs-induced cytotoxicity. CEHs were dispersed from 14-day-old chicken embryo livers and exposed to Aroclor 1254 (A1254) in the range of 0.1-10 microg/ml, A1254 (10 microg/ml) and each vitamin (10 microg/ml) for 24 h. Cell viability was evaluated by determinations of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) leakage. The antioxidant status, namely cellular lipid peroxidation, was evaluated by measuring the thiobarbituric acid reactive substances (TBARS) and glutathion (GSH) levels and superoxide dismutase (SOD) activities. The cultured CEHs maintained normal polygonal cell shape and formed confluent monolayer after 24-h culture. A1254 (10 microg/ml) caused irreversible damage to cell membrane integrity and induced cell death in a dose-dependent manner. It induced increased TBARS production, decreased SOD activity and GSH concentration. VE, VC and VA alone or combinations of VE+VC and VE+VA significantly attenuated A1254-induced toxic effects, which suggested that lipid peroxidation was involved in the sequence of events leading to A1254-induced damage or death of the cultured CEHs. These results indicated that CEHs in serum-free culture represented a suitable model for rapid toxicity assessment of environmental pollutants such as PCBs in a visible manner. Antioxidant vitamins displayed protective effects on CEHs from A1254-induced damage through preventing lipid peroxidation. PMID- 15896555 TI - Correlation of in vitro cytotoxicity with paracellular permeability in Caco-2 cells. AB - This in vitro study aims to develop a cell culture model that compares paracellular permeability (PP) with acute cytotoxicity (AC). Caco-2 cells were seeded in 96-well plates and on polycarbonate filter inserts. Confluent monolayers were exposed to increasing concentrations of 20 reference chemicals for 24-h and 72-h. Cytotoxicity was determined using MTT and NRU cell viability assays in 96-well plates. PP was measured using transepithelial electrical resistance (TEER) measurements, as well as passage of lucifer yellow (LY), [3H] mannitol (both low mw indicators), and FITC-dextran (higher mw indicator) in culture inserts. Inhibitory concentrations 50% (IC50s) suggest that there were good correlations between 24-h and 72-h exposures. NRU IC50 values correlated better with TEER, which is consistent with the Registry of Cytotoxicity (RC; ICCVAM) database report. Both cell viability assays indicate that cytotoxicity occurs before TEER is compromised. In addition, 24-h and 72-h NRU assays, and 72 h TEER measurements, displayed the highest correlations with established rodent LD50s. PP experiments showed that passive paracellular transport of the tight junction markers, especially [3H]-mannitol, correlates with the IC50s determined with the viability assays and TEER measurements. Our AC/TEER/PP model thus allows for the differentiation between the concentrations necessary for AC and those needed to interfere with PP. We propose that the in vitro AC, TEER and PP results be used to compute a formula which can "normalize" and improve the predictive ability of in vitro acute cytotoxicity assays for in vivo lethality. PMID- 15896556 TI - Symptom validity assessment: practice issues and medical necessity NAN policy & planning committee. AB - Symptom exaggeration or fabrication occurs in a sizeable minority of neuropsychological examinees, with greater prevalence in forensic contexts. Adequate assessment of response validity is essential in order to maximize confidence in the results of neurocognitive and personality measures and in the diagnoses and recommendations that are based on the results. Symptom validity assessment may include specific tests, indices, and observations. The manner in which symptom validity is assessed may vary depending on context but must include a thorough examination of cultural factors. Assessment of response validity, as a component of a medically necessary evaluation, is medically necessary. When determined by the neuropsychologist to be necessary for the assessment of response validity, administration of specific symptom validity tests are also medically necessary. PMID- 15896557 TI - The Ruff Figural Fluency Test: heightened right frontal lobe delta activity as a function of performance. AB - Research has indicated that the Ruff Figural Fluency Test [RFFT; Ruff, R. M., Light, R. H., & Evans, R. W. (1987). The Ruff Figural Fluency Test: A normative study with adults. Developmental Neuropsychology, 3, 37-51] is sensitive to right frontal lobe functioning. Indeed, research has differentiated between patients with left or right frontal lobe lesions using performance on the RFFT [Ruff, R. M., Allen, C. C., Farrow, C. E., Niemann, H., & Wylie, T. (1994). Figural fluency: Differential impairment in patients with left versus right frontal lobe lesions. Archives of Clinical Neuropsychology, 9, 41-55]. The present investigation used quantitative electroencephalography to test further whether the RFFT was sensitive to right frontal lobe functioning among a group of individuals with no history of head injury. To meet this objective, the RFFT was administered to a group of 45 right-handed men with no history of significant head injury or cerebral dysfunction. Delta magnitude (muV) at three right frontal electrode sites (FP2, F4, F8) was then used to compare those who performed the best (High Fluency) with those who performed the worst (Low Fluency) on the RFFT. The findings indicated heightened right frontal delta magnitude for the Low Fluency group relative to the High Fluency group at the F2 and F8 right frontal electrode sites. Thus, the present findings provide further support for the contention that the RFFT is sensitive to right frontal lobe functioning, even among those with no history of head injury. PMID- 15896558 TI - Alternate-form reliability of the Dementia Rating Scale-2. AB - The Dementia Rating Scale-2 (DRS-2) is a frequently used assessment of cognitive status among older adults in both research and clinical practice. Despite its well-established psychometric properties, its use in serial assessments has posed limitations with regard to practice effects. The primary purpose of the present study is to provide preliminary evidence of alternate-form reliability for the DRS-2. A heterogeneous sample of 52 community-dwelling adults over age 60 with no reported diagnosis of dementia were administered the DRS-2 as well as a newly developed alternate form [DRS-2: AF; Schmidt, K. S. (2004). Dementia Rating Scale 2 Alternate Form: Manual supplement. Lutz, FL: Psychological Assessment Resources]. Our results reveal strong correlations between the two forms; further, no significant differences were found between total scale and subscale scores obtained from the two forms. Therefore, the DRS-2: AF may be a valuable assessment tool in both research and clinical arenas. PMID- 15896559 TI - The impact of specific language impairment on working memory in children with ADHD combined subtype. AB - The objective of this study was to examine the impact of comorbid specific language impairment (SLI) on verbal and spatial working memory in children with DSM-IV combined subtype Attention Deficit Hyperactivity Disorder (ADHD-C). Participants were a clinical sample of 812- to 1212-year-old children diagnosed with ADHD-C. A group of ADHD-C with SLI was compared to a group of ADHD-C without SLI, and a group of normal children, matched on age and nonverbal intelligence. The results show that ADHD-C children with SLI scored significantly lower than those without SLI and normal children, on verbal working memory measures only. Both ADHD groups performed normally on spatial working memory measures. It is concluded that working memory deficits are not a specific characteristic of ADHD but are associated with language impairments. The importance of screening for language disorders in studies of neuropsychological functioning in children with ADHD is emphasized. PMID- 15896560 TI - ADHD subtypes: do they differ in their executive functioning profile? AB - The present study was designed to investigate the hypothesis that children with Attention Deficit Hyperactivity Disorder combined subtype (ADHD-C) have a generalized executive functioning (EF) [Barkley, R. A. (1997). Behavioural inhibition, sustained attention, and executive functions: Constructing a unifying theory of AD/HD. Psychological Bulletin, 121, 65-94; Barkley, R. A. (1997). ADHD and the nature of self-control. New York: The Guilford Press]. We tested whether ADHD-C and ADHD inattentive subtype (ADHD-I) can be differentiated from each other on EF measures. We compared 16 normally developing boys with 16 boys with ADHD-C and 16 with ADHD-I on five EF domains. The boys were all matched on age, IQ, and the presence of oppositional defiant disorder (ODD)/conduct disorder (CD). Despite carefully diagnosed groups and methodological controls, the results do not support the EF-hypothesis of ADHD-C. Children with ADHD-C differed from normal controls (NC) on tasks related to inhibition; they did not exhibit EF deficits on all EF tasks. Children with ADHD-C also exhibited deficits on non-EF tasks. Furthermore, the ADHD-C and ADHD-I subtypes did not differ from one another. Neuropsychological findings on the domains under study did not yield evidence for the distinctiveness of ADHD-C and ADHD-I subtypes. PMID- 15896561 TI - Norming subjects for the Halstead Reitan battery. AB - The adequacy of neuropsychological norms is dependent on the characteristics of the subjects whose data are used. Volunteer participants and neurologically normal participants in major normative studies show markedly different patterns of test performance that reflects on the inadequacy of using volunteer participants to develop norms. In this study when all of the Halstead Reitan Battery norming studies with an N of 200 or more were examined, Wechsler FSIQ score differences of approximately 1 standard deviation above average for the volunteer normative participants was found. The norms from the norming study using neurologically normal patients were essentially average. The exclusion of neurologically suspect participants from volunteer normative studies lead to a bias reflected in artificially inflated levels of performance, and a restricted range of variation. This sets inappropriately rigid assessment cutoffs for defective performance. Furthermore, data collection from neurologically normal subjects follows the same format as that used in the assessment of neurologically compromised subjects, while the more research oriented protocol used to collect normative data from volunteer participants does not. PMID- 15896562 TI - Test-retest reliable coefficients and 5-year change scores for the MMSE and 3MS. AB - The present study explored several different procedures for determining the amount of change that occurred on the Mini-Mental State Exam [MMSE; Folstein, M. F., Folstein, S. E., & McHugh, P. R. (1975). "Mini-Mental State": A practical method for grading the cognitive state of patients for the clinician. Journal of Psychiatric Research, 12, 189-198] and Modified Mini-Mental State Exam [3MS; Teng, E. L., & Chui, H. C. (1987). The Modified Mini-Mental State (3MS) examination. Journal of Clinical Psychiatry, 48, 314-318] over short and extended test-retest intervals. The test-retest scores were drawn from a selected sample of elderly individuals who participated in the Canadian Study of Health and Aging [Canadian Study of Health and Aging. (1994). The Canadian study of health and aging: Study methods and prevalence of dementia. Canadian Medical Association Journal, 150, 899-913] and were tested on two occasions (CSHA-1 and CSHA-2) separated by 5 years. On each occasion the MMSE and 3MS were administered twice at approximately 3-month intervals. Thus, the mental status tests were administered four times: times 1 and 2 at CSHA-1 and times 3 and 4 at CSHA-2. Mean difference scores and percent of baseline scores showed relatively small group changes over both short and long test-retest intervals for the MMSE and the 3MS. A reliable change index based on a linear regression model controlled for practice effects, psychometric errors due to low reliability, regression to the mean, and accounted for the effects of various demographic variables. Consequently, this reliable change index provided a better estimate of the amount of change that occurred for individual participants than did the mean Retest-Test 1 difference, percent of baseline change, or a reliable change index based on a Retest-Test 1 difference score. Normative data for the change scores are provided. PMID- 15896563 TI - Cognitive reserve and anosognosia in questionable and mild dementia. AB - Cognitive reserve (CR) theory posits that the clinical presentation of individuals with the same brain disease varies based upon premorbid variables (e.g., education, occupation, reading ability). Anosognosia (decreased insight regarding one's deficits) is common in dementia and has implications for safety, treatment, and caregiver burden. The current study examined the role of CR in anosognosia in individuals with mild dementia. Participants were individuals diagnosed with questionable or mild dementia (Clinical Dementia Rating 0.5 or 1) after neuropsychological evaluation. Anosognosia was measured by informant patient discrepancy on the Cognitive Difficulties Scale. High and Low CR groups were created based upon reading performance. Low CR showed greater anosognosia than High CR. Anosognosia was associated with reduced reading performance, even after controlling for global cognitive decline. These findings suggest CR is related to anosognosia in questionable and mild dementia, and have clinical implications for the assessment of awareness in dementia. PMID- 15896564 TI - Practice effects and the use of alternate forms in serial neuropsychological testing. AB - Accurate understanding of practice characteristics, performance stability, and error on neuropsychological tests is essential to both valid clinical assessment and maximization of signal detection for clinical trials of cognitive enhancing drugs. We examined practice effects in 28 healthy adults. As part of a larger study using donepezil and simulating a Phase I trial, participants were randomized into: placebo, no-treatment and donepezil. Donepezil results are presented elsewhere. Neuropsychological tests were administered in a fixed order for 6 weeks, with alternate forms available for most tests. Despite alternate forms, ANOVAs revealed significant improvements for the pooled control group (placebo and no-treatment) on all tests except Letter Number Sequencing and Trails B. Learning occurred principally in the first three to four sessions. PASAT and Stroop interference showed the greatest learning. Thus, serial assessment with alternate forms may attenuate retest effects on some tests, but continued learning occurs on novel tests or those in which an advantageous test taking strategy can be identified. Alternate forms and baseline practice sessions may help control early, rapid improvements in clinical trials. PMID- 15896565 TI - Analysis of the WAIS-III Index score scatter using the significant deviation from the mean of the four Index scores. AB - Despite their validity and their potential clinical interest, the WAIS-III Index scores seem less used by clinicians than the traditional IQs. One of reason of this lack of interest in the Indices is the relative complexity of their interpretation. We advocate the use of the average index score as a baseline to analyze the Index score variability and to identify strengths and weaknesses in the profile of the Index scores. Davis's formula provides reference values to be used to highlight the Index scores significantly moving away from the baseline. The usefulness of this method is illustrated with the analysis the Index score scatter in the standardization sample of the French adaptation of the WAIS-III. A rather important dispersion of the Index scores around the average index score was observed, a large percentage of standardization sample showing one or two Index scores significantly different from the baseline. PMID- 15896566 TI - Anxious-depression in boys: an evaluation of executive functioning. AB - The effects of anxiety and depression on frontal lobe functioning were tested in two groups of 9-11-year-old boys. Participants were screened for handedness, health, intelligence and classified as anxious-depressed or non-anxious, non depressed based on scores from the A-State scale of the State-Trait Anxiety Inventory for Children and the Child Depression Inventory. Previous research in our laboratory has indicated that boys high in anxious-depression may have neuropsychological deficits [e.g., Emerson, C. S., Harrison, D. W., & Everhart, D. E. (1999). Investigation of receptive affective prosodic ability in school aged boys with and without depression. Neuropsychiatry, Neuropsychology and Behavioral Neurology, 12(2), 102-109; Emerson, C. S., Harrison, D. W., Everhart, D. E., & Williamson, J. B. (2001). Grip strength asymmetry in depressed boys. Neuropsychiatry, Neuropsychology, and Behavioral Neurology, 14(2), 130-134]. In order to assess the effects of anxious-depression on cerebral functioning performance on the Trail Making Test (Forms A and B) and on the Concept Formation subtest of the Woodcock Johnson was compared between groups. As predicted, anxious-depressed boys demonstrated deficits in sequencing, alternation, and problem-solving tasks as evidenced by longer completion times and significantly more errors on the tests. These results provide supportive evidence for deficits in frontal lobe functioning. PMID- 15896567 TI - The relation of self-report of mood and anxiety to CVLT-C, CVLT, and CVLT-2 in a psychiatric sample. AB - The relation between mood and cognitive status has been examined extensively over the years suggesting a significant impact of mood and potentially anxiety on memory. This relation is of particular interest to practitioners conducting evaluations in settings that regularly treat individuals diagnosed with psychiatric disorders. With this in mind, the present study sought to evaluate the impact of self-report of depression and anxiety on the California Verbal Learning Test-children's version (CVLT-C), the California Verbal Learning Test (CVLT), and the California Verbal Learning Test-2nd edition (CVLT-2) in a mixed psychiatric sample. Records from 107 patients aged 6-78 evaluated in an outpatient psychiatry unit were examined. Results indicated minimal predictive utility was provided from self-report symptoms of anxiety or depression on CVLT C, CVLT, or CVLT-2: Trial 1, Trial 5, total score, Short Delay Free Recall, Short Delay Cued Recall, long delay free recall, or long delay cued recall performance above the variance predicted by age, gender, and Full Scale IQ. Additional variance predicted by depression as measured by the Beck Depression Inventory (BDI) and the Child Depression Inventory (CDI) or anxiety as measured by the State-Trait Anxiety Inventory (STAI) was less than 3.0% over that accounted for by the covariates for the great majority of measurements from the various CVLT versions. Exceptions included the CDI that tended to account for approximately 5.0% of the variance on all of the CVLT-C measures and the STAI that accounted for approximately 5.0-10.0% additional variance on the short and long delay measures of the CVLT-2. The present results suggest that performance on the various forms of the CVLT are minimally predicted by self-reported depression and anxiety in a psychiatric setting. PMID- 15896568 TI - Modified Wisconsin Card Sorting Test: proposal of a supplementary scoring method. AB - To assess categorizing ability, we propose a new scoring criterion for the MCST, the "categorizing efficiency", taking into account the number of cards used by the subject to complete a maximum of six categories. The advantage of adding that parameter to traditional ones is evaluated in a small population of normal children and adults and those affected with pathologies. PMID- 15896569 TI - A model of smooth pursuit in primates based on learning the target dynamics. AB - While the predictive nature of the primate smooth pursuit system has been evident through several behavioural and neurophysiological experiments, few models have attempted to explain these results comprehensively. The model we propose in this paper in line with previous models employing optimal control theory; however, we hypothesize two new issues: (1) the medical superior temporal (MST) area in the cerebral cortex implements a recurrent neural network (RNN) in order to predict the current or future target velocity, and (2) a forward model of the target motion is acquired by on-line learning. We use stimulation studies to demonstrate how our new model supports these hypotheses. PMID- 15896570 TI - Information theory, novelty and hippocampal responses: unpredicted or unpredictable? AB - Shannon's information theory provides a principled framework for the quantitative analysis of brain responses during the encoding and representation of event streams. In particular, entropy measures the expected uncertainty of events in a given context. This contextual uncertainty or unpredictability may, itself, be important for balancing [bottom-up] sensory information and [top-down] prior expectations during perceptual synthesis. Using event-related functional magnetic resonance imaging (fMRI), we found that the anterior hippocampus is sensitive to the entropy of a visual stimulus stream. In contrast, activity in an extensive bilateral cortico-thalamic network was dictated by the surprise or information associated with each particular stimulus. In short, we show that the probabilistic structure or context in which events occur is an important predictor of hippocampal activity. PMID- 15896572 TI - Global exponential stability of delayed competitive neural networks with different time scales. AB - A competitive neural network model was recently proposed to describe the dynamics of cortical maps, where there are two types of memories: long-term and short-term memories. Such a network is characterized by a system of differential equations with two types of variables, one models the fast neural activity and the other models the slow modification of synaptic strength. In this paper, we introduce a time delay parameter into the neural network model to characterize the signal transmission delays in real neural systems and the finite switch speed in the circuit implementations of neural networks. Then, we analyze the global exponential stability of the delayed competitive neural networks with different time scales. We allow the model has non-differentiable and unbounded functions, and use the nonsmooth analysis techniques to prove the existence and uniqueness of the equilibrium, and derive a new sufficient condition ensuring global exponential stability of the networks. PMID- 15896575 TI - The dynamic wave expansion neural network model for robot motion planning in time varying environments. AB - We introduce a new type of neural network--the dynamic wave expansion neural network (DWENN)--for path generation in a dynamic environment for both mobile robots and robotic manipulators. Our model is parameter-free, computationally efficient, and its complexity does not explicitly depend on the dimensionality of the configuration space. We give a review of existing neural networks for trajectory generation in a time-varying domain, which are compared to the presented model. We demonstrate several representative simulative comparisons as well as the results of long-run comparisons in a number of randomly-generated scenes, which reveal that the proposed model yields dominantly shorter paths, especially in highly-dynamic environments. PMID- 15896577 TI - Predictive neural networks for gene expression data analysis. AB - Gene expression data generated by DNA microarray experiments have provided a vast resource for medical diagnosis and disease understanding. Most prior work in analyzing gene expression data, however, focuses on predictive performance but not so much on deriving human understandable knowledge. This paper presents a systematic approach for learning and extracting rule-based knowledge from gene expression data. A class of predictive self-organizing networks known as Adaptive Resonance Associative Map (ARAM) is used for modelling gene expression data, whose learned knowledge can be transformed into a set of symbolic IF-THEN rules for interpretation. For dimensionality reduction, we illustrate how the system can work with a variety of feature selection methods. Benchmark experiments conducted on two gene expression data sets from acute leukemia and colon tumor patients show that the proposed system consistently produces predictive performance comparable, if not superior, to all previously published results. More importantly, very simple rules can be discovered that have extremely high diagnostic power. The proposed methodology, consisting of dimensionality reduction, predictive modelling, and rule extraction, provides a promising approach to gene expression analysis and disease understanding. PMID- 15896578 TI - Intraoperative transesophageal echocardiography: standard monitor and diagnostic instrument for difficult situations? PMID- 15896579 TI - CD11b may be a less satisfactory indicator for cardiac ischemia-reperfusion injury in coronary artery bypass graft surgery with cardiopulmonary bypass than cardiac troponin I. AB - STUDY OBJECTIVE: To investigate whether CD11b on neutrophils can be used as a marker to predict myocardial ischemia-reperfusion injury for patients undergoing coronary artery bypass graft (CABG) surgery on cardiopulmonary bypass (CPB). DESIGN: Prospective, observational study. SETTING: University teaching hospital and university hospital laboratories. PATIENTS: Twelve adults, physical status II and III (American Society of Anesthesiologists) patients scheduled for elective CABG surgery on CPB. INTERVENTIONS: Anesthesia and surgical interventions were performed according to standard procedures. Coronary sinus was cannulated for retrograde cardioplegia perfusion and for coronary sinus blood sampling. The blood samples were collected from systemic and coronary venous sinus blood respectively at several time points before CPB, during CPB, and after declamping of the ascending aorta. MEASUREMENTS AND MAIN RESULTS: CD11b expressions in systemic circulation blood increased significantly during CPB and maintained higher levels after cardiac reperfusion (P < .05); on the other hand, CD11b expressions in coronary sinus blood declined gradually and reached their lowest level at 5 minutes after aortic declamping. The differences in CD11b expressions between systemic circulatory blood and coronary sinus blood after cardiac reperfusion were significant (P < .05). Cardiac troponin I (cTnI) concentrations in both systemic circulatory blood and coronary sinus blood increased significantly after cardiac reperfusion (P < .05), and the concentrations of cTnI in coronary sinus blood increased much higher than the corresponding concentrations of cTnI at each time point (P < .05). CONCLUSION: CD11b expression on neutrophils may not be a reliable predictor for myocardial ischemia reperfusion injury in CABG surgery on CPB because of the possible sequestration of neutrophils in myocardium. PMID- 15896580 TI - The intubating laryngeal mask airway: rocuronium improves endotracheal intubating conditions and success rate. AB - STUDY OBJECTIVE: To assess intubating conditions without neuromuscular blocking drugs, to determine the relation between the dose of rocuronium and the probability of achieving excellent or at least good (good or excellent) intubating conditions with the intubating laryngeal mask airway (ILMA), and finally, to determine the relationship between rocuronium use and the success rate of endotracheal intubation. DESIGN: Prospective, randomized, double-blinded, placebo-controlled study. SETTING: University-affiliated medical center. PATIENTS: Sixty American Society of Anesthesiologists physical status I and II patients undergoing elective surgery. INTERVENTIONS: Anesthesia was induced with propofol 2.5 mg/kg and fentanyl 1 microg/kg. One minute after loss of consciousness, patients received rocuronium 0.2 mg/kg or saline. In the rocuronium group, if intubating conditions were scored as poor, rocuronium dose in the next patient was increased by 0.05 mg/kg. If intubating conditions were scored as good, no change was made, but if conditions were scored as excellent, the dose was decreased by 0.05 mg/kg. One minute after rocuronium or saline administration, an ILMA was used to intubate the trachea. If intubation was unsuccessful, a second attempt was made using the ILMA. MEASUREMENTS: We recorded intubating conditions and the success rate of tracheal intubation. MAIN RESULTS: Without rocuronium, the probability of achieving at least good intubating conditions with the ILMA was 30%. A rocuronium dose of 0.2 mg/kg resulted in a probability of 80% to achieve at least good intubating conditions. Rocuronium significantly increased the success rate of the second intubation attempt. CONCLUSION: To achieve good or excellent intubating conditions with the ILMA, a rocuronium dose lower than the standard intubating dose of 0.6 mg/kg can be used. Neuromuscular blockade increases the success rate of intubation if a second attempt is necessary. PMID- 15896581 TI - Addition of 0.1% bupivacaine to buprenorphine and droperidol in patient controlled epidural analgesia improved postoperative pain scores on coughing after gynecological surgery. AB - STUDY OBJECTIVE: To compare the analgesic efficacy of additional 0.1% bupivacaine to patient-controlled epidural analgesia (PCEA) using buprenorphine and droperidol after gynecological surgery. DESIGN: Randomized, double-blinded study. SETTING: Operating theater and general ward at Jichi Medical School Hospital. PATIENTS: Thirty patients with American Society of Anesthesiologists physical status I and II scheduled for gynecological surgery. INTERVENTIONS: Patients received combined general and epidural anesthesia for surgery and epidural analgesia for postoperative analgesia. Patients were assigned to receive PCEA with or without 0.1% bupivacaine. Group 1 (n = 15) received buprenorphine 20 microg and droperidol 0.1 mg diluted with saline, and group 2 (n = 15) received bupivacaine 2 mg, buprenorphine 20 microg, and droperidol 0.1 mg diluted with saline (0.1% bupivacaine solution) in a bolus dose of the PCEA, respectively. No background epidural infusion was used. MEASUREMENTS: Visual analog pain scale (VAPS) scores at rest and on coughing, and cumulative frequency of self administrated analgesic solution in PCEA were recorded at 24 and 48 hours postoperatively. MAIN RESULTS: There were no significant differences noted between the groups in VAPS scores at rest or in cumulative volumes of PCEA solution in 24 or 48 hours postoperatively. Median VAPS scores on coughing in group 2 were significantly lower than those values in group 1 at 24 hours (36 vs 65 mm, P < .001) and 48 hours (32 vs 54 mm, P = .036) postoperatively. CONCLUSIONS: Addition of 0.1% bupivacaine to PCEA using buprenorphine and droperidol provides better analgesia on coughing after gynecological surgery. PMID- 15896582 TI - High mortality rate in patients with advanced liver disease independent of exposure to general anesthesia. AB - STUDY OBJECTIVE: To evaluate the survival of patients with advanced liver disease to determine if known exposure to general anesthesia within a 5-year period has a measurable effect on mortality. DESIGN: Retrospective survival analysis of male veterans with advanced liver disease. SETTING: Tertiary referral VA Medical Center and university-affiliated teaching hospital. MEASUREMENTS: One hundred twenty-seven patients with a history of alcoholic cirrhosis and documented hepatitis C infection and stable platelet counts were identified and then divided into 3 groups. The 5-year survival rates in all 3 groups were compared using Kaplan-Meier survival curves. MAIN RESULTS: Ninety patients had marked thrombocytopenia (<100000/mm3). Their survival rates with and without known exposure to general anesthesia were compared with those of control subjects with alcoholic cirrhosis and hepatitis C infection but with platelet counts greater than 100000/mm3. The 5-year survival rate of 57% in the group that received general anesthesia was comparable to the 58% rate observed in the group without this exposure. Both groups' rates were statistically lower than the 5-year survival rate of 77% in the group with advanced liver disease but without thrombocytopenia. CONCLUSION: Comparably high mortality rates were observed in patients with advanced liver disease with or without exposure to general anesthesia. Higher survival rates were noted in patients with advanced liver disease who were not thrombocytopenic. PMID- 15896583 TI - Human middle cerebral artery flow velocity during controlled hypotension combined with hemodilution-transcranial Doppler study. AB - STUDY OBJECTIVE: To evaluate the effects of controlled hypotension combined with hemodilution on human middle cerebral artery flow velocity (Vmca) by transcranial Doppler ultrasonography. DESIGN: Randomized prospective study. SETTING: Inpatient surgery at Nagasaki Rosai Hospital. PATIENTS: Thirty American Society of Anesthesiologists physical status I and II patients scheduled for total hip arthroplasty. INTERVENTIONS: Anesthesia was maintained with nitrous oxide-oxygen (N(2)O-O(2)) and sevoflurane during normocapnia. Hemodilution was carried out after induction of anesthesia, in which blood was withdrawn then replaced with the same amount of hydroxyethyl starch to achieve a final hematocrit level of 32% (group A = mild hemodilution group, N = 15) or 24% (group B = moderate hemodilution group, N = 15). In both groups, controlled hypotension was induced with prostaglandin E(1) to maintain mean arterial pressure at approximately 55 mm Hg for 80 minutes. MEASUREMENTS AND MAIN RESULTS: Vmca and blood gas were measured before hemodilution, after hemodilution, 80 minutes after starting hypotension, and 60 minutes after recovery from hypotension. Vmca significantly increased in group A (+122%) and group B (+156%) after each hemodilution. In group B, Vmca was significantly greater than baseline values at 80 minutes after starting hypotension (+135%) and 60 minutes after recovery from hypotension (+140%). CONCLUSION: The combination of moderate hemodilution, such as hematocrit value of 24%, and prostaglandin E(1)-induced hypotension would not impair middle cerebral artery flow during sevoflurane-N(2)O-O(2) anesthesia during normocapnia. PMID- 15896584 TI - Reduction of postoperative emetic episodes and analgesic requirements with dexamethasone in patients scheduled for dental surgery. AB - STUDY OBJECTIVE: To evaluate the antiemetic and analgesic effects of dexamethasone for the first 24 hours postoperatively in patients scheduled for dental surgery. DESIGN: Prospective, randomized, double-blind, placebo-controlled study. SETTING: University hospital. PATIENTS: One hundred twenty ASA physical status I and II patients (45 men, 75 women; aged 17-48 years) undergoing general anesthesia for dental surgery. INTERVENTIONS: Patients received intravenously placebo or dexamethasone at 3 different doses (4, 8, or 16 mg) at the end of the surgical procedure. A standard general anesthetic technique, including sevoflurane and nitrous oxide in oxygen, was used. MEASUREMENT AND MAIN RESULTS: Emetic episodes and analgesic requirements were evaluated. The rate of patients who were emesis free (no nausea, retching, or vomiting) during 0 to 24 hours after anesthesia was 63% with dexamethasone 4 mg (P = NS), 83% with dexamethasone 8 mg (P < .05), and 90% with dexamethasone 16 mg (P < .05), compared to placebo (60%). The need for diclofenac sodium for intolerable pain was less in patients who had received dexamethasone 8 mg or dexamethasone 16 mg than in those who had received placebo or dexamethasone 4 mg (P < .05). No clinically important adverse events were observed in any group. CONCLUSIONS: Prophylactic dexamethasone 8 mg is effective for the prevention of nausea and vomiting after dental surgery and in the management of postoperative pain. Increasing the dose to 16 mg provides no further benefit. PMID- 15896585 TI - A comparison of fentanyl-propofol with a ketamine-propofol combination for sedation during endometrial biopsy. AB - STUDY OBJECTIVE: The purpose of this study was to compare the clinical activities of ketamine and fentanyl when used in combination with propofol for outpatients undergoing endometrial biopsy. The investigated parameters were respiration, sedation, recovery rate, side effects, time to discharge, and patient satisfaction. DESIGN: Prospective, randomized, double-blind study. SETTING: University hospital. PATIENTS: Forty American Society of Anesthesiologists physical status I and II patients (age range 38-61 years) scheduled for elective endometrial biopsy. INTERVENTIONS: Patients received intravenous bolus doses of either fentanyl 1 microg/kg and propofol 1 mg/kg in (group 1 n = 20) or ketamine 0.5 mg/kg and propofol 1 mg/kg in (group 2 n = 20). MEASUREMENTS: Heart rate, systolic and diastolic blood pressure, respiratory rate, and peripheral O(2) saturation were monitored in all patients. Depth of sedation was assessed by the Ramsay sedation score. The Aldrete score was used for postoperative recovery evaluation. The time to Aldrete score >/=8, side effects, and time to discharge were recorded. MAIN RESULTS: Respiratory depression was observed in 5 patients from group 1 and 1 patient from group 2, but the difference was not significant (P > .05). Nausea, vertigo, and visual disturbances were more frequent in group 2 (P < .05). The time to Aldrete score >/=8 was similar in the 2 groups (6.2 +/- 3.5 minutes in group 1 and 7.0 +/- 3.1 minutes in group 2); the time to discharge was significantly longer in group 2 (71.2 +/- 9.7 minutes in group 1 and 115.2 +/ 25.6 minutes in group 2). Frequency of patient satisfaction was 95% in group 1 and 60% in group 2. CONCLUSIONS: Hemodynamic change and degrees of sedation showed that fentanyl-propofol and ketamine-fentanyl combinations can be used safely in patients undergoing endometrial biopsy. However, with regard to side effects and patient satisfaction, the fentanyl-propofol was superior. PMID- 15896586 TI - Preoperative beta-blocker use: is titration to a heart rate of 60 beats per minute a consistently attainable goal? AB - STUDY OBJECTIVE: To quantify the prevalence of perioperative beta-blocker use and its impact on preoperative and preinduction heart rate (HR), in light of the recent publication of specific recommendations regarding perioperative beta blocker use and desired HR. DESIGN: Retrospective observational study in patients who underwent elective and coronary artery bypass graft (CABG) surgery between January 2001 and March 2002. SETTING: Tertiary-care teaching hospital. MEASUREMENTS: Percentage of eligible patients who received beta-blockers preoperatively and the impact of non-protocol-based beta-blocker therapy on preadmission and preinduction HR were recorded. Differences were assessed with unpaired t test and chi(2) analysis; P < .05 was considered significant, with corrections for multiple comparisons. RESULTS: Of the patients who underwent vascular surgery, 9 had documented prior beta-blocker intolerance. Of the remaining 172 patients, 94.8% had indication for perioperative beta-blocker use. However, only 47.7% of the eligible patients received beta-blockers. Of the 155 CABG patients, 74.2% were taking beta-blockers preoperatively. Only 29% of vascular patients and 32% of CABG patients who were receiving beta-blockers had HR less than 60 beats per minute (bpm) at preadmission. The mean preadmission HR in vascular surgery patients was 65.2 +/- 11 and 73.2 +/- 13.8 bpm in beta blocker and non-beta-blocker patients, respectively (P = .0001). In CABG surgery patients, preadmission HR values were 64.2 +/- 13 and 76.1 +/- 12 bpm in beta blocker and non-beta-blocker patients, respectively (P = .001). The preinduction HR subsequently increased in the beta-blocker as well as in the non-beta-blocker groups. CONCLUSION: Only half of the patients who qualify to receive preoperative beta-blockers by current recommendations actually receive them before noncardiac surgery, and the majority of these patients have preadmission and preinduction HR less than 60 bpm. Targeting beta-blocker therapy treatment to an HR less than 60 bpm may not be readily achievable in many patients. PMID- 15896587 TI - Anesthetic management for transurethral resection of the bladder in a 74-year-old man with uncorrected tetralogy of Fallot. AB - We report a case of general anesthesia for transurethral resection of the bladder in a 74-year-old male patient with uncorrected tetralogy of Fallot. This case illustrates the pertinent pathophysiology of the complex cardiac lesion related to tetralogy of Fallot as well as the feasibility and issues with regard to the anesthetic management. PMID- 15896588 TI - Systemic vasodilation is a predominant cause of hypotension in a patient with familial amyloid polyneuropathy during liver transplantation. AB - A patient with familial amyloid polyneuropathy underwent a living, nonrelated orthotopic liver transplant and developed hypotension after induction of anesthesia. Causes of hypotension in patients with familial amyloid polyneuropathy are discussed. Transesophageal echocardiographic monitoring was invaluable in differentiating various causes of hypotension and in diagnosing peripheral vasodilation as the predominant cause of hypotension. PMID- 15896589 TI - Cardiovascular collapse during elective orthopedic surgery: massive intraoperative pulmonary thromboembolism treated with emergent cardiopulmonary bypass. AB - Successful management strategies for pulmonary thromboembolism-primarily published as case reports-include a spectrum ranging from medical treatment with cardiovascular support and anticoagulation to more invasive interventions such as pulmonary embolectomy. We present a case of massive intraoperative pulmonary embolism that was managed with emergent pulmonary embolectomy. Unlike previously reported cases, this aggressive management strategy was unsuccessful, due in part perhaps to lack of discernible clot during embolectomy. PMID- 15896590 TI - Delayed pneumomediastinum and pneumothorax complicating laparoscopic extraperitoneal inguinal hernia repair. AB - A 53-year-old healthy man underwent elective laparoscopic, extraperitoneal, right sided herniorrhaphy. Postoperatively, he complained of chest pain on inspiration. Chest x-ray and computed tomographic scan revealed a pneumomediastinum and a right-sided pneumothorax. Previous case reports and possible etiologies are reviewed. PMID- 15896591 TI - Massive pulmonary thromboembolism during elective spine surgery. AB - Massive pulmonary thromboembolism (PTE) is a condition that can still be seen in the operating room despite the use of thromboprophylaxis. A high degree of suspicion of this condition is necessary to achieve an early diagnosis and a rapid treatment to improve patient outcome. We report on a 27-year-old patient who sustained a massive PTE while undergoing a second-stage anterior release and posterior fusion of his thoracolumbar spine for idiopathic scoliosis. PMID- 15896592 TI - A case of suspected severe pulmonary embolism in a living-related liver transplantation donor. AB - Optimal safety for a donor is an essential condition for living-related liver transplantation (LRLT). Severe pulmonary embolism may be a rare complication for the donors, but it is potentially fatal. Various risk factors and prophylactic methods for thromboembolic complications have been described. We report here a case of severe pulmonary embolism in a living-related liver transplantation donor who underwent right liver lobectomy, although the patient had no apparent risk factor for thromboembolism except for slight obesity (body mass index = 27). In addition, the donor received prophylactic use of graduated compression stockings and intermittent pneumatic compression by use of a special device from the start of anesthesia until the patient was able to ambulate. Fortunately, the patient was successfully treated with urokinase and heparin infusion followed by warfarin administration. PMID- 15896593 TI - Neostigmine-induced prolonged neuromuscular blockade in a patient with atypical pseudocholinesterase. AB - A patient with unrecognized atypical pseudocholinesterase was given succinylcholine and then vecuronium before neuromuscular monitoring was instituted. Subsequently, when neostigmine and glycopyrrolate were given to reverse what was thought to be a nondepolarizing block, the patient became further relaxed, and his trachea could not be extubated for more than 10 hours. In this report, we discuss drug interactions, phase II block, and the importance of timely neuromuscular monitoring. PMID- 15896594 TI - Changing of an anesthesiology clinical base year to create an integrated 48-month curriculum: experience of one program. AB - To allow for growth in our anesthesiology residency, we assumed control of the clinical base year (postgraduate year 1[PGY-1]) and adjusted the curriculum to accommodate the expanded size. With this opportunity to change the curriculum, we created a clinical base year to prepare PGY-1 for clinical anesthesia training in PGY-2 to PGY-4 using, for this purpose, the best resources of our clinical site. We describe the process and preliminary results of the change. PMID- 15896595 TI - Ultrasound guidance for epidural catheter placement: a coming of age? PMID- 15896596 TI - Identification of the obscured trachea using the Trachlight. PMID- 15896597 TI - Middle ear barotrauma in general anesthesia: special care. PMID- 15896598 TI - Surgical outcomes in congenital heart disease: expectations and realities. AB - The past 50 years of congenital heart surgery has produced enormous progress. Current results surpass expectations. Yet there are important residual problems in patients growing up after heart surgery for congenital heart disease. Our system of care must evolve to care for these people throughout their lives. The evolution of congenital heart surgery has reached a point in time when we should extend care to patients in under serviced emerging countries. Development of local expertise will be required within those countries that are willing to commit resources to an organized program of caring for people with congenital heart disease. Database technology is an essential tool for ensuring and improving quality of care in every congenital heart centre. Both Registry and Academic databases have much to offer in improving care for future patients. Yet overzealous privacy laws threaten the knowledge base provided by computerized databases. We need to guide our legislators in ensuring that the valuable resource provided by database technology is not lost. PMID- 15896599 TI - Evidence for palliative enlargement of the right ventricular outflow tract in severe tetralogy of Fallot. AB - OBJECTIVE: If the pulmonary artery (PA) tree in patients with Fallot's tetralogy (TOF) is extremely hypoplastic, a shunt procedure may be difficult and not desirable because of side-effects. Moreover, the failing catch-up growth of the pulmonary annulus is well known. In patients with a severe form of TOF, we performed palliative transannular patching of the right ventricular outflow tract. The early and long-term follow-up was evaluated. METHODS: Eleven patients (93 days (10-245 days); 3.5+/-0.7 kg (2.5-4.3 kg)) had highly symptomatic TOF (Hb: 18+/-2g/dl, SO2: 68+/-11%); angiographic diameters: RPA: 4.1 mm (2.5-6.4 mm), LPA: 3.4 mm (1.6-7.0 mm), PA trunc: 4.4 mm (2.5-7.0 mm). All 11 underwent transannular enlargement of the right ventricular outflow tract without closure of the ventricular septum defect. A PA index (cross-sectional area of the pulmonary arteries to BSA) was used to compare pre- and postoperative data. For follow-up, the patients were repetitively examined clinically and echocardiographically. RESULTS: Preoperative PA index was 87+/-40 mm2/m2 (normal: 330+/-35 mm2/m2). Postpalliation angiograms (age: 10-14 months) demonstrated a significant catch-up growth in nine patients (PA index from 99+/-40 to 310+/-54 mm2/m2) and inadequate growth in two patients (PA index 63 and 115 mm2/m2). Perioperative mortality was zero. Ten patients (43 months; 6-105 months) underwent elective repair. Six patients received pulmonary homograft valves (6-15 years after repair) because of severe pulmonary valve insufficiency and severe RV dilation. COMPLICATIONS: One patient died 10 months postpalliation due to pneumonia, one patient received a pacemaker after repair and died (2 months post repair) due to pacemaker failure, a 5-year-old patient died 1 month after repair due to sepsis. All eight long-term survivors (12-17 years) are in excellent clinical condition. Echocardiography revealed good RV function and near normal diameters at peak systolic pressures between 25 and 50 mmHg. Only one patient developed brady-arrhythmia; a pacemaker was implanted 8 years after repair and 2 years after homograft implantation. CONCLUSIONS: In a very severe form of TOF, palliative right ventricular outflow tract construction may provide the potential for complete repair. In the presented high-risk patient group, mortality was not related to the hypoplastic pulmonary arteries. Obviously, all patients need pulmonary valve implantation in the long run. PMID- 15896600 TI - Staged surgical repair of functional single ventricle in infants with unobstructed pulmonary blood flow. AB - OBJECTIVE: The infant with a functional single ventricle (SV) and unobstructed pulmonary blood flow (UPBF) requires early protection of the pulmonary vascular bed to ensure suitability for a subsequent Fontan procedure. Systemic obstruction by aortic arch obstruction, subaortic stenosis, or combination of both, has been widely recognized as an important risk factor for poor outcome in children with SV-UPBF who are palliated with pulmonary artery banding (PAB). We reviewed our experience with primary PAB in the subset of patients with SV-UPBF to identify risk factors for subsequent palliative procedures and Fontan completion. METHODS: Between January 1990 and May 2004, 80 patients (median age, 14 days) with functional SV and UPBF underwent PAB as their primary palliative procedure. Thirty-five neonates had concomitant aortic coarctation or interrupted aortic arch repair (44%). A Damus-Kaye-Stansel procedure was subsequently performed in 19 patients, and subaortic resection or ventricular septal defect or bulboventricular foramen enlargement was performed in five. RESULTS: There were 4 operative deaths, and 15 late deaths. The actuarial overall survival is 84% at 1 year, 76% at 5 and 15 years. Follow-up is complete in all but six children at a mean interval of 4.9+/-3.7 years (range, 2 months-15 years). Thirty-seven patients (49%; 37 of 76) have undergone the hemi-Fontan procedure (with three hospital deaths) and 40 patients (53%; 40 of 76; 12 children without previous hemi-Fontan) have undergone the completion Fontan procedure without mortality or Fontan takedown. CONCLUSION: In infants with single ventricle physiology with or without systemic outflow obstruction and unobstructed pulmonary blood flow, a strategy of pulmonary artery banding carries acceptable operative and mid-term mortality in a high-risk group of patients. Pulmonary artery banding does not compromise performance of subsequent Damus-Kaye-Stansel procedure or completion Fontan palliation. PMID- 15896601 TI - Right ventricle-to-pulmonary artery shunt and modified Blalock-Taussig shunt in preparation to hemi-Fontan procedure in children with hypoplastic left heart syndrome. AB - OBJECTIVE: The advantageous effect of right ventricle-to-pulmonary artery shunt (RV-PA) on the early postoperative hemodynamics in the Norwood procedure for hypoplastic left heart syndrome (HLHS) is well known. Numerous controversies still exist with respect to the late consequences of this new palliation method in preparation for the second stage procedure. METHODS: Between September 1997 and September 2004, a consecutive series of 78 children with HLHS from a single institution underwent the hemi-Fontan procedure: Group 1 (n=27) after Blalock Taussig shunt (BT), and Group 2 (n=51) after RV-PA. Hemodynamic, echocardiographic and clinical perioperative data were analyzed. RESULTS: There were no significant differences in the age and operative weight (Group 1: 6.9+/ 1.04 months, 6.22+/-0.99 kg; Group 2: 6.57+/-1.12 months, 6.36+/-0.86 kg). Children after RV-PA were characterized by a significantly higher preoperative hematocrit value (P=0.014), lower aortic and superior vena cava oxygen blood saturation (P<0.001, P=0.024), severe right ventricle hypertrophy more rarely diagnosed in echocardiography (P<0.004), lower Qp:Qs ratio (P=0.011), larger right (P=0.001) and left (P=0.006) pulmonary artery index and a shorter intensive care unit stay after the hemi-Fontan procedure (P=0.004). CONCLUSIONS: The Norwood procedure with the RV-PA shunt provides satisfactory late hemodynamics and improves the development of the pulmonary arteries. Children with hypoplastic left heart syndrome subjected to this new method of palliation are good candidates for the hemi-Fontan procedure. PMID- 15896602 TI - Restrictive left atrial outflow adversely affects outcome after the modified Norwood procedure. AB - OBJECTIVE: Moderate restrictive foramen ovale in neonates with hypoplastic left heart syndrome (HLHS) is considered to be favourable, reducing pulmonary overcirculation, before modified Norwood operation. However, some newborns with severe restriction of interatrial communication will have pulmonary vascular disease at birth, which correlates with increased perioperative mortality. This article studies the post-Norwood hemodynamic patterns and outcome for the particular group of HLHS newborns with restrictive left atrial outflow compared to other patients. METHODS: Restrictive left atrial outflow is defined as mitral and/or aortic atresia with intact ventricular septum, and restrictive foramen ovale, with 3 mm diameter or less with mean interatrial pressure gradient more than 5 mmHg at preoperative echo-Doppler. Four neonates fulfilled these criteriae among 18 consecutive patients, who underwent Norwood procedure from October 2002 to December 2003. Mean arterial pressure, heart rate, mean common atrial pressure, urinary output, central venous and arterial oximetry data, serum lactate levels, and dosages of milrinone, phentolamine and norepinephrine were collected at 0, 6, 12, 18 and 24 h after operation. Data were summarized as mean+/-SEM. For univariate comparison of different variables, Student's t-test was used. RESULTS: The postoperative hemodynamic pattern of patients with restrictive left atrial outflow was characterized by hypoxemia and low cardiac output. Arterial (66+/-3.0% vs 76+/-1.0%, P=0.01) and central venous (37+/-1.2 vs 52+/-1.1%, P=0.001) oxygen saturations were much lower than in patients without restriction. Arterio-venous oxygen saturation difference was wider (29+/-2.4% vs 23+/-0.9%, P=0.02) and serum lactate levels were higher (10.8+/-3.0 vs 2.8+/-0.2 mmol/l, P=0.03). Common atrial pressures were more elevated (12+/-0.8 vs 8+/-0.3 mmHg, P<0.001) and higher norepinephrine doses were needed (0.44+/-0.15 vs 0.06+/ 0.01 microg/kg/min, P=0.03). The difference for the mean arterial pressures did not reach the significance level (48+/-2.0 vs 51+/-2.0 mmHg, P=0.2). Operative mortality was higher 75% (3/4) compared to 14.3% (2/14, P=0.04) for the other patients. CONCLUSIONS: Restrictive left atrial outflow adversely affects outcome after modified Norwood procedure. Abnormal pulmonary vasculature leading to insufficient pulmonary perfusion is incriminated. To improve outcome, implantation of larger size modified Blalock-Taussig or right ventricle-to pulmonary artery shunts and routine use of postoperative mechanical assist device should be considered. PMID- 15896603 TI - Comparison of hemodynamics between Norwood procedure and systemic-to-pulmonary artery shunt for single right ventricle patients. AB - OBJECTIVE: Despite that surgical outcomes of patients with hypoplastic left heart syndrome have improved, one of the problems remaining is the high interstage mortality after a stage I Norwood procedure. The purpose of this study was to determine the hemodynamic characteristics of hypoplastic left heart syndrome after a Norwood procedure. We examined the perioperative hemodynamic differences of the staged operation between the first stage of the Norwood procedure and systemic pulmonary shunt for single right ventricle patients. METHODS: Data from 39 patients who underwent a Norwood procedure (right ventricle to pulmonary artery conduit: 19, Blalock-Taussig shunt, 20) were analyzed. There were nine early and seven interstage deaths. Bidirectional cavopulmonary shunt was performed in 15 patients and the Fontan procedure in 9 (group H). We defined the control group as 26 patients who underwent the first stage of a systemic pulmonary shunt for a single ventricle. Bidirectional cavopulmonary shunt was performed in 14 patients and the Fontan procedure in 8 (group C). We compared the perioperative hemodynamics of the staged operation between the two groups. RESULTS: Cardiothoracic ratio and single ventricular diastolic dimension before bidirectional cavopulmonary shunt were acutely increased in group H (P=0.02, <0.001). There was no significant difference between the two different types of Norwood procedures. The pulmonary artery index for the right heart bypass operation was lower in group H than in group C (P<0.001). Oxygen saturation before bidirectional cavopulmonary shunt in group H decreased (P<0.001) and thus was lower than that in group C (P=0.003). Mortality and the postoperative clinical parameters of the right heart bypass operation were not different between the two groups. CONCLUSIONS: Patients with hypoplastic left heart syndrome showed hemodynamic instability of acutely increased cardiothoracic ratio, and single ventricular diastolic dimension despite decreased oxygen saturation interstage after stage I of a Norwood procedure. This suggests that this hemodynamic characteristics in hypoplastic left heart syndrome correlates with the higher mortality before second stage palliation than in found with single right ventricle patients. PMID- 15896604 TI - Results of the Ross operation in a pediatric population. AB - OBJECTIVE: To analyse the results of the mid-term clinical and echocardiographic follow-up of the pediatric Ross operation. METHODS: Echo-Doppler follow-up of 53 consecutive pediatric Ross procedures performed between 1994 and 2003. Median age was 9.7 years at time of operation (2 weeks-17.7 years). Six patients were younger than 3 months. Median age at follow-up was 15.6 years. Aortic valve/left ventricular outflow tract (LVOT) anomalies were congenital in 49 (92%). Seventy percent had previous surgery or balloon valvuloplasty. Root replacement was used in all. Thirteen patients (25%) had LVOT enlargement. Mean cross-clamp time was 113 (69-189) minutes. RESULTS: Early mortality occurred in 3 patients after emergency surgery following balloon failure (n=1) and extended Ross following interrupted arch/VSD repair (n=2). Late mortality was due to LV fibroelastosis in 2 patients and complicated pulmonary artery stenting in another. RVOT reoperations were required because of late homograft obstruction in 2 patients and because of pulmonary artery stenosis in another. Five patients (9.4%) were reoperated for pulmonary autograft dilatation (n=3) and for leaflet fibrosis or perforation (n=2). Autografts were repaired in two patients, while a mechanical valve was inserted in 3 cases. At 9 years the actuarial survival and event free survival were 89 and 74%, respectively. At last follow-up 90% of autograft diameters indexed to body surface area was above the 90th percentile of normal aortic root diameters. LVOT and RVOT gradients were low and autograft insufficiency was trivial to mild in 84% and mild to moderate in 16%. Autograft stenosis was not noticed. CONCLUSIONS: The pediatric Ross procedure remains an important tool but autograft dilatation also occurs in the pediatric population. The significance of this finding has yet to be determined. PMID- 15896605 TI - Structure function interface with sequential shortening of basal and apical components of the myocardial band. AB - OBJECTIVE: To mechanically test the intact cardiac structure to determine the sequence of contraction within the myocardial mass to try to explain ejection and suction. METHODS: In 24 pigs (30-85 kg), segment shortening at the site of sonomicrometer crystals was continuously recorded. The ECG evaluated rhythm, and Millar pressure transducers measured intraventricular pressure and dP/dt. RESULTS: Study of segment shortening defined a sequence of contraction within the myocardial mass, starting at the free wall of the right ventricle and on the endocardial side of the antero-septal wall of the left. Crystal location defined underlying contractile trajectory; transverse in right ventricle followed by basal posterior left ventricle, and from the endocardial anterior wall to the posterior apical segment and finally to the epicardial side of the anterior wall. Mean shortening fraction averaged 18+/-3%, with endocardial exceeding epicardial shortening by 5+/-1%. Epicardial segment crystal displacement followed endocardial shortening by 82+/-23 ms in the anterior wall, and finished 92+/-33 ms after endocardial shortening stopped, time frame that matches the interval of fast drop of ventricular pressure and the start of suction. CONCLUSIONS: Crystal shortening fraction sequence followed the rope-like myocardial band model to contradict traditional thinking, with two starting points of excitation contraction, the right anterior free wall of the right ventricle, and the endocardial side of the anterior wall. Active suction may be due to active shortening of the epicardial fibers of the anterior wall, because relaxation was not detected when both mitral and aortic valves were closed during the interval previously termed 'isovolumetric relaxation'. PMID- 15896606 TI - Intraoperative assessment of right ventricular volume and function. AB - OBJECTIVE: Right ventricular function is an important aspect of global cardiac performance which affects patients' outcome after cardiac surgery. Due to its geometrical complexity, the assessment of right ventricular function is still a very difficult task. Aim of this study was to investigate the value of a new technique for intraoperative assessment of right ventricle based on transesophageal 3D-echocardiography, and to compare it to volumetric thermodilution by using a new generation of fast response thermistor pulmonary artery catheters. METHODS: Twenty-five patients with coronary artery disease underwent 68 intraoperative measurements by 3D-echocardiography and thermodilution simultaneously. Following parameters were analysed: right ventricular end-diastolic volume (RVEDV), end-systolic volume (RVESV) and ejection fraction (RVEF). Pulmonary, systemic and central venous pressures were simultaneously recorded. Segmentation of right ventricular volumes were obtained by the 'Coons-Patches' technique, which was implemented into the EchoAnalyzer, a multitask system developed at our institution for three-dimensional functional and structural measurements. RESULTS: Right ventricular volumes obtained by 3D echocardiography did not show significant correlations to those obtained by thermodilution. Volumetric thermodilution systematically overestimates right ventricular volumes. Significant correlations were found between RVEF measured by 3D-echocardiography and those obtained by thermodilution (r=0. 93; y=0.2+0.80x; SEE=0.03; P<0.01). Bland-Altmann analysis showed that thermodilution systematically underestimates RVEF. The bias for measuring RVEF was +15.6% with a precision of +/-4.3%. The patients were divided into two groups according to left ventricular function. The group of patients with impaired function showed significantly lower right ventricular ejection fraction (44.1+/-4.6 vs. 55.1+/ 3.9%; P<0.01). CONCLUSIONS: Three-dimensional echocardiography provides a useful non-invasive tool for intraoperative and serial assessment of right ventricular function. This new technique, which overcomes the limitations of previous methods, may offer key insights into management and outcome of patients with severe impairment of cardiac function. PMID- 15896607 TI - Impact of postoperative blood pressure control on regression of left ventricular mass following valve replacement for aortic stenosis. AB - OBJECTIVE: Considerable left ventricular (LV) hypertrophy sometimes remains after aortic valve replacement (AVR) for aortic stenosis. For this issue, most previous studies have focused solely on transprosthetic pressure gradient, although true problem is not the pressure gradient itself but an elevated LV pressure. This study investigated the impact of blood pressure on postoperative LV mass regression, which had been overlooked in previous studies. METHODS: Seventy-nine adult patients with pure aortic stenosis who were treated with AVR using bileaflet mechanical valves underwent echocardiography before surgery, around 6 months later ('early'), and 2-3 years later (31.7+/-14.7 months, 'late'). Patients were divided into two groups whether postoperative systolic blood pressure was below (n=47; N group) or above 130 mmHg (n=32; H group) following recommendation of WHO-ISH and JNC 7th report. Preoperative LV mass (g/m2) did not differ significantly (232+/-80 vs. 243+/-76, P=0.91). RESULTS: LV mass became significantly smaller and regression was significantly more effective in N group than in H group both at 'early' (145+/-43 vs. 180+/-54, regression against preoperative value 34.6+/-19.1 vs. 19.9+/-26.6%, P=0.007) and 'late' (132+/-41 vs. 178+/-51, regression 41.1+/-16.0 vs. 21.0+/-27.0%, P<0.001) evaluations. Regression between 'early' and 'late' evaluations was significant only in N group (P=0.012). The LV mass index returned to the normal range at 'late' evaluation in 52.1% of N group and 12.5% of H group patients (P<0.001), and 25 out of 29 patients without residual LV hypertrophy were N group patients. Multivariate analyses revealed that preoperative LV mass index (P<0.001) and postoperative systolic blood pressure (P=0.007) showed significant influence on postoperative LV mass index, and postoperative systolic blood pressure alone significantly (P<0.001) influenced the regression ratio of the LV mass against the preoperative value. No prosthesis related variables (size, orifice area index, pressure gradient) had significant influence. CONCLUSIONS: For LV mass regression after AVR, postoperative blood pressure appeared to be more important than prosthesis selection. Controlling the systolic blood pressure below 130 mmHg was beneficial, which coincided with recommendation of WHO-ISH and JNC 7th report despite the pressure drop due to prosthesis in the aortic position. PMID- 15896608 TI - Right ventricular disarticulation. An 18-year single centre experience. AB - OBJECTIVE: Right ventricular disarticulation (RVD) is an accepted procedure in the treatment of ventricular tachycardia of right ventricular origin. We set out to review the long-term outcomes with RVD at our institution for patients with arrhythmogenic right ventricular dysplasia (ARVD) or refractory tachycardia. A renewed interest in this operation has come about in patients unable to tolerate implantable cardioverter defibrillators. METHODS: Seventeen patients had RVD carried out between 1985 and 2003. There were 15 males and 2 females. The age range was 14-72 (median: 34). Six patients had partial RVD and 11 a complete RVD. ARVD was confirmed in 15 patients at histology. Biventricular pacing was used post-operatively in the two most recent patients. RESULTS: The follow-up was complete in 94% (16/17). The median follow-up was 13 years (0-18). The overall hospital mortality was 6% (1/17). Over the follow-up period there were three deaths 9, 11 and 17 years post-surgery. Heart transplantation due to biventricular failure was required in two patients. In the group followed up for more than 10 years the over all event free survival at 10 years was 77% (3/13). CONCLUSIONS: In cases of refractory ventricular tachycardia, where multiple antiarrhythmic medication, repeated catheter ablation and ICD insertion are unsuccessful at symptom control, RVD is an excellent antiarrhythmic procedure. In the long term, signs of biventricular failure present, possibly, dependent on the natural history of ARVD. The long-term effect of biventricular pacing on the disarticulated right ventricle is yet to be defined. PMID- 15896609 TI - The Dor procedure for left ventricular reconstruction. Ten-year clinical experience. AB - OBJECTIVE: Surgical ventricular restoration by means of the Dor procedure is a surgical option in patients with coronary artery disease, postinfarction left ventricular aneurysm or ischemic dilated cardiomyopathy with or without ventricular tachycardia. The aim of this study was to evaluate the 10-year clinical experience of this procedure in our institution. METHODS: From May 1994 to June 2004, surgical ventricular restoration was performed in 101 patients (77 males), mean age 63.8 (35-80) years. All patients presented with angina and/or heart failure and/or ventricular tachycardia. Postinfarction left ventricular aneurysm was present in 97 patients and ischemic dilated cardiomyopathy with a large akinetic left ventricle in 4. The preoperative left ventricular ejection fraction was 27+/-10 (7-50) %. Multi-vessel disease was present in 80 patients. Ventricular tachycardia was diagnosed in 53 patients (spontaneous VT in 25). Mitral regurgitation more than grade 2 was found in 13 patients. The mean Euroscore was 7.1+/-2.9 (3-17). RESULTS: All patients underwent the Dor procedure, which in 53 patients included a non-guided endocardectomy and cryoablation for treatment of ventricular tachycardia. Coronary artery bypass grafting was performed in 99 patients and a mitral valve procedure was performed in 29. Intra-aortic balloon pumping was used postoperatively in 14 cases and 24 patients needed inotropic support for more than 24 h. Mean time on the ventilator was 16+/-25 (3-168) hours and mean stay in the intensive care unit was 2.1+/-2.2 (0-13) days. Postoperative stroke occurred in 7 patients. Early mortality was 8/101 (7.9%). Mean follow-up in operative survivors was 4.4+/-2.8 (0.1-10.4) years. Actuarial survival at 1, 3 and 5 years was 88, 79 and 65%. CONCLUSIONS: The Dor procedure is a reproducible surgical option for treatment of postinfarction left ventricular aneurysm. Early and long-term results are good in terms of survival. PMID- 15896610 TI - Downsizing of the mitral valve and coronary revascularization in severe ischemic mitral regurgitation results in reverse left ventricular and left atrial remodeling. AB - OBJECTIVE: Data of combined mitral downsizing by restrictive prosthetic ring annuloplasty and coronary artery bypass grafting (CABG) in patients with ischemic cardiomyopathy and moderately severe to severe mitral regurgitation (MR) are rare, and little is known about the effect on reverse left ventricular (LV) and left atrial (LA) remodeling. METHODS: Thirty-eight patients (70.6+/-8.3 years) with coronary artery disease, ischemic cardiomyopathy (LV ejection fraction [LVEF] 31+/-8%) and moderately severe to severe MR (grade 3.6+/-0.5) underwent CABG and mitral downsizing by 2-4 ring sizes. Clinical follow-up and serial transthoracic echocardiographic studies were performed after surgery (discharge, 3+/-0.5 months, 13+/-7 months) to assess survival, NYHA class, MR, leaflet coaptation height, LA and LV dimensions/volumes, fractional shortening (FS) and LVEF. RESULTS: Early mortality (<30 days) was 2.6%, survival at follow-up was 92 and 85%, respectively. NYHA class improved from 3.3+/-0.6 to 1.5+/-0.6 (P<0.001). Residual MR at discharge and at follow-up was grade 0.5 and 0.6, respectively (P<0.001). Leaflet coaptation height was 8+/-1 mm and did not change over time. LV end-diastolic, end-systolic and LA dimensions decreased from 60+/-7 to 57+/-8 mm, from 47+/-9 to 42+/-9 mm and from 51+/-5 to 45+/-4 mm, FS increased from 23+/ 9 to 28+/-10% (P<0.001); LV end-diastolic and end-systolic volumes decreased from 188+/-33 to 171+/-30 ml and from 129+/-35 to 105+/-33 ml, LVEF increased from 31+/-8 to 39+/-10% (P<0.001). CONCLUSIONS: Combined mitral downsizing and CABG surgery was performed with excellent clinical results: only minimal residual MR, a significant reduction of LA dimension and an increase of LV contractility due to reverse remodeling were observed. PMID- 15896611 TI - Intramyocardial microdepot injection increases the efficacy of skeletal myoblast transplantation. AB - OBJECTIVE: Recent progress in the field of cellular cardiomyoplasty has opened new prospects for the treatment of ischemic heart disease and currently moves from bench to bedside. The aim of the present study was to develop a novel cell delivery technique, reducing target tissue damage and improving cell dispersion and engraftment. METHODS: In 30 male Fischer F344 rats an infarction of the left ventricle was generated by ligation of the left anterior descendent artery. Seven days after infarction, either 15 microdepots of 10 microl myoblast cell suspension (microdepot group) or culture medium (control group) were injected into the infarcted region using an automatic pressure injection device, or three depots of 50 microl myoblast cell suspension (macrodepot group) were injected using the standard surgical technique. Echocardiography was performed in all rats before and 6 weeks after cell injection. In all groups the perioperative mortality was below 20%. Six weeks after cell transplantation, a significant improvement of ejection fraction was seen in both myoblast treated groups compared to controls (macrodepot, microdepot, control; 53.7+/-11.9, 70.7+/-2.0, 39.1+/-6.4; P=0.026, P<0.001). The microdepot group showed a more decent improvement than the macrodepot group (70.7+/-2.0 vs. 53.7+/-11.9, P=0.013). In both treated groups, grafted myoblasts differentiated into multinucleated myotubes within host myocardium, however, the engraftment pattern was different and angiogenesis was enhanced in the microdepot group. CONCLUSIONS: Intramyocardial multisite pressure injection allows the safe and reliable transplantation of several myoblast microdepots into an infarcted myocardium and improves the efficacy of myoblast transplantation compared to the standard technique. PMID- 15896612 TI - Cumulative risk adjusted monitoring of 30-day mortality after cardiothoracic transplantation: UK experience. AB - OBJECTIVE: Guidelines are needed for real-time quality monitoring in heart and lung transplantation. The cumulative sum (CUSUM) methodology with boundary limits derived from the sequential probability ratio test (SPRT) provide a means of monitoring performance without the need for repeated statistical testing. The variable life adjusted display (VLAD) complements the SPRT chart and provides a directly interpretable assessment of performance. We present our experience with these charts in monitoring cardiothoracic transplant outcomes in the UK. METHODS: Thirty-day-post-transplantation mortality after isolated first time transplantation of the heart (n=1634) or lung (n=1162) in adults, between July 1995 and March 2004 in eight centres were monitored. CUSUM charts, with and without risk-adjustment and risk-adjusted VLAD plots were constructed. Thirty-day mortality rates for the UK as a whole were taken as the reference values for the unadjusted charts and a 50% increase in risk provided the basis for construction of boundary lines for the SPRT. Risk-adjustment was based on multivariate models previously developed from the national database. RESULTS: For heart transplantation without risk-adjustment, four centres crossed the lower boundary, indicating 30-day mortality was in-line with or better than seen nationally. Two centres were close to signalling an alert, warning of a rise in mortality rate, but neither chart signalled an alarm. After risk-adjustment one centre's graph moved towards the centre of the chart, indicating monitoring should continue and the other signalled an alarm. For lung transplantation the unadjusted mortality rate at one centre was confirmed acceptable and the results remained inconclusive for five. At the other centre, following an alert to a possible increase in mortality half-way through the sequence, results improved. Case-mix adjustment served to pull the charts away from the upper boundary lines; no chart suggested any cause for concern. For most centres the VLAD charts oscillated around the horizontal axis. CONCLUSIONS: CUSUM charts are useful tools for monitoring performance, and provide a basis for visually comparing results between centres and identifying periods of 'bad runs'. Risk-adjustment, which down-weights higher risk activity, avoids inappropriate reaction to unadjusted breaches of alert and alarm lines. PMID- 15896613 TI - Synergistic effect of low dose cyclosporine A and human interleukin 10 overexpression on acute rejection in rat lung allotransplantation. AB - OBJECTIVE: Electroporation mediated transfer of plasmid DNA into peripheral muscle results in high transfection efficiency. The aim of this study was to investigate the effect of gene transfer of human IL-10 (hIL-10) into the tibialis anterior muscle (MTA) in combination with low dose Cyclosporine A (CsA) on acute rejection of lung allografts in the rat. METHODS: Lung allotransplantation was performed from male BN donor to male Fisher F344 rats. Gene transfer was achieved by intramuscular injection into the MTA of the recipient followed by electroporation (4 x 20 ms impulses at 200 V/cm) 24 h prior to the transplantation. Group A (n=5) received CsA (2.5 mg/kg bw ip) for 5 days post transplant and group B (n=5) 2.5 microg of PCIK hIL-10 (plasmid expression vector containing human CMV immediate early gene promoter and enhancer) and a low dose CsA (2.5 mg/kg bw i.p.). Graft function was assessed by blood gas at day 5 after exclusion of the native lung. Animals were sacrificed and blood was drawn to measure serum hIL-10 levels (ELISA) and tissue was sampled for histological grading of rejection. RESULTS: Local expression of hIL-10 was confirmed at the mRNA level by in situ hybridization. All group A control animals showed severe signs of rejection. At day 5 all grafts in group B showed good gas exchange mean PaO2 233+/-123 mmHg, vs 44+/-8 mmHg in group A. Histological examination revealed moderate to severe rejection in all animals in group A (IIIB, ISHLT) in contrast to low moderate rejection in group B (II-IIIA). hIL-10 serum levels on day 5 were 14+/-7 pg/ml in group B vs. 0 in group A. CONCLUSIONS: Electroporation mediated hIL-10 overexpression in a peripheral muscle of the recipient in combination with low dose CsA reduces acute rejection in this model of rat lung allotransplantation. PMID- 15896614 TI - Fetal origins of coronary heart disease-implications for cardiothoracic surgery? AB - Over the last 15 years, there has been growing evidence that poor nutrition during gestation plays an important role in the development of coronary heart disease. This hypothesis, commonly known as the 'fetal origins of adult disease' has now gained widespread acceptance in the scientific community. In this review, we discuss the evidence for this theory and analyse the patho-physiological mechanisms underlying the relationship between altered fetal growth and coronary heart disease. Finally, the potential relevance of the theory to cardiac surgical practice will be evaluated. PMID- 15896615 TI - Unstable angina and non-ST segment elevation: surgical revascularization with different strategies. AB - OBJECTIVE: Unstable angina/non-ST elevation myocardial infarction (UA/NSTEMI) still causes significant hospital morbidity and mortality. We evaluated whether surgical outcome can be modified by different myocardial protection strategies. METHODS: This was a prospective clinical study conducted in the cardiac surgery units of two university hospitals. Two hundred and sixty-two consecutive patients undergoing CABG for UA/NSTEMI between January 2002 and June 2004 were prospectively divided in three groups: 126 patients underwent on-pump CABG with antegrade blood cardioplegia (Group A); 67 underwent antegrade and retrograde blood cardioplegia (Group B); 69 off-pump CABG (Group C). Hospital outcome was analysed. Differences in outcome variables were detected with ANOVA; Tukey's multiple comparison test and Tamhane's T2 test were used when appropriate. RESULTS: Group A showed higher mortality (P=.001; P=.014 vs. Group B; P=.003 vs. Group C) and perioperative myocardial infarction (P=.001; P=.016 vs. Group B; P=.05 vs. Group C). Hospital stay was shorter in Group B and Group C, compared to Group A (P=.005; P=.043 and P=.05, respectively). Group A required higher doses of inotropes compared to Group B and Group C (P=.0001; P=.0001 and P=.03, respectively), whereas Group B and Group C did not require any inotropic support at all (P=.0001; P=.002 and P=.001 vs. Group A, respectively). Total morbidity was higher in Group A (P=.006; P=.007 vs. Group B; P=.005 vs. Group C). Wall motion score index recovered only in Group B (P=.0001) and Group C (P=.001). Troponin I was higher in Group A at 12 h (P=.0001; P<.001 vs. Group B and Group C), 24 (P=.0001; P=.001 vs. Group B and Group C), 48 (P=.0001; P=.001 vs. Group B, P=.002 vs. Group C) and 72 h (P=.0001; P=.004 vs. Group B; P=.05 vs. Group C). CONCLUSIONS: Isolated antegrade cardioplegia should be questioned in UA/NSTEMI. Outcome using off-pump revascularization was as good as that of combined antegrade and retrograde warm blood cardioplegia. PMID- 15896616 TI - Preoperative statin use and in-hospital outcomes following heart surgery in patients with unstable angina. AB - OBJECTIVE: Unstable angina (UA) is characterized by a state of coronary artery vascular inflammation and endothelial dysfunction. Statins mitigate inflammation and endothelial dysfunction and decrease mortality associated with percutaneous interventions for UA. We determined whether preoperative statin use is associated with decreased mortality and morbidity following coronary artery bypass+/-valve surgery for UA. METHODS: Patients with CCS Class IV angina having CABG+/-valve surgery were identified (n=1706). A logistic regression model determined the association of preoperative statin use with in-hospital mortality (IHM). Propensity score analysis was used to match two sub-groups of patients (GrpI, on statins, n=534; GrpII, not on statins, n=534) on factors known to affect outcomes. Outcomes were IHM, intra-aortic balloon pump (IABP) use, perioperative myocardial infarction (PMI), prolonged (>24 h) ventilation (p-vent), stroke, and a composite outcome (comp) defined as any one or more of the above. RESULTS: Of the 1706 patients, 1075 were on statins and 631 were not. Patients on statins were more likely to have isolated CABG, EF>40%, and be on a beta-blocker (P=0.0001); and less likely to have renal failure, MI<7 days, CHF, and undergoing urgent/emergent surgery (P=0.0001). Unadjusted rates of IHM (9 vs. 5%, P=0.001), stroke (4.4 vs. 2.3%, P=0.015), p-vent (28.4 vs. 19%, P=0.0001), and comp (32.5 vs. 22.8%, P=0.0001) were lower in patients receiving statins. After adjustment, statin use was not associated with a reduction in IHM (OR=1.0, 95% CI=0.6-1.5, P=0.85) or comp (OR=1.1, 95% CI=0.8-1.4, P=0.69). No significant differences were found in any of the propensity-adjusted outcomes for GrpI vs. GrpII: IHM (7.1 vs. 6.4%), PMI (2.8 vs. 1.7%), IABP use (3 vs. 3.8%), stroke (3.8 vs. 3.9%), p-vent (26.4 vs. 23.8%), comp (31.5 vs. 27.5%). CONCLUSIONS: Preoperative statin use is not associated with a reduction in IHM or major morbidity following CABG+/-valve surgery in patients with UA. PMID- 15896617 TI - On-pump beating heart versus off-pump coronary artery bypass surgery-evidence of pump-induced myocardial injury. AB - OBJECTIVE: By maintaining native coronary blood flow in on-pump beating heart surgery (OnP-BH) and comparing with OPCAB strategy pump-related effects on myocardial injury and cardiac dysfunction could be specifically differentiated from ischemia/reperfusion-related consequences of surgical coronary revascularization. METHODS: In a randomized-prospective design, 40 elective patients with normal EF and three vessels coronary artery disease (left main disease excluded) were assigned to OPCAB or OnP-BH surgery. Before coronary occlusion and 1, 30, 60, and 90 min after reperfusion with the LIMA graft, coronary sinus (CS) blood was sampled to determine intraoperative myocardial ischemia (pH, lactate, pO2) and oxidative stress (malondialdehyde, MDA). Additionally to CS blood arterial blood was analyzed 4, 12, and 24 h postoperatively to determine myocardial necrosis (CK-MB, cardiac troponin I), myocardial dysfunction (NT-proBNP) and inflammation (C-reactive protein). RESULTS: Groups were identical with regards to age and gender (OPCAB 63.0+/-6.0 versus OnP-BH 65.3+/-3.9 y, 20% female patients). Number of grafts were 3.0+/-0.5 in OPCAB versus 2.9+/-0.3 in OnP-BH (n.s.) with 44 versus 34% bilateral IMAs and 56 versus 50% complete arterial revascularization. Regarding ischemia, intraoperatively only lactate values increased significantly in the OnP-BH group. Significantly higher CK-MB and troponin I levels were found from LIMA-LAD flow release onwards to 4 h postoperatively in the OnP-BH group. NT-proBNP levels were significantly higher in the OnP-BH group during the entire study period. CRP levels were higher in the OnP-BH group 12 and 24 h postoperatively. CONCLUSIONS: In this randomized study on routine coronary patients with normal ventricular function, OPCAB revealed less myocardial injury than OnP-BH. These findings implicate that CPB slightly affects the myocardium. PMID- 15896618 TI - Skeletonized versus pedicled internal mammary artery: impact of surgical technique on post CABG surgery pain. AB - OBJECTIVE: Recent evidence suggests that coronary artery bypass graft (CABG) surgery often results in chronic chest wall pain, termed: 'post CABG pain syndrome' (PCP). Direct injury to intercostal nerves during the surgical procedure was presumed to underlie this syndrome. The aim of this study was to investigate the effect of two harvesting techniques of the internal mammary artery (IMA)-skeletonization (S-LIMA) and pedicle (P-LIMA) on the occurrence and incidence of PCP. METHODS: A mailed questionnaire enquiring about the presence and characteristics of PCP was sent to all 482 patients who had undergone CABG in our institution in the years 1999-2000. A randomly chosen subgroup of IMA patients reporting PCP were summoned for evaluations of pain localization and intensity, thermal and tactile sensitivity, and disability assessment using recognized tests and indices. RESULTS: Of the 380 responders (S-LIMA: 221, P LIMA: 125, veins only V-34) 169 (44%) reported having PCP. Its prevalence was similar between the two IMA groups (S-LIMA: 45%, P-LIMA: 50%) but significantly lower in the V group (18%, P=0.003). Physical assessment in the subgroup of 43 IMA patients (S-LIMA: 22, P-LIMA: 21) performed 40.2+/-8.7 months after surgery confirmed equal occurrence of mostly left and midline chest wall neuropathic pain in both IMA groups. No significant differences were found between the two groups in respect to indices of pain intensity, thermal and tactile sensitivity and disability. CONCLUSIONS: PCP is a prevalent finding in post CABG patients. The skeletonization technique of IMA harvesting although causing significantly less inner chest wall trauma does not appear to reduce the occurrence of PCP. This finding may imply that ischemic injury rather than direct mechanical injury to the intercostal nerves is the putative mechanism underlying PCP. PMID- 15896619 TI - Clinical experience with the Novare Enclose II manual proximal anastomotic device during off-pump coronary artery surgery. AB - OBJECTIVE: The aim of this study was to assess the feasibility and safety of a new proximal anastomotic device during off-pump coronary surgery. METHODS: The Novare Enclose II is a manual proximal anastomotic device that enables the surgeon to perform proximal anastomoses without side-biting clamps in a fully pressurized aorta. The device was used in 30 off-pump coronary artery bypass graft (CABG) patients (Group A) for performing 25 vein, 10 free right internal thoracic artery and 14 radial artery anastomoses to the aorta. The number of proximal anastomoses varied between 1 and 3 per patient. Intraoperative transcranial Doppler measurements were performed to detect cerebral microemboli during the use of the device and were compared with transcranial Doppler measurements of 30 off-pump patients where a partial side clamp was used for proximal anastomoses (Group B). The aorta was evaluated using transesophageal echocardiography (TEE) and epiaortic echocardiography (EAE) in both groups before the procedure and patients with a calcified aorta were excluded. RESULTS: In Group A, 49 proximal anostomoses were performed as planned with the device using conventional hand-sewn methods. Twenty-four of these were arterial conduits. In Group B, 32 proximal anastomoses were performed using 25 veins and 7 arterial grafts. There were no procedure related to adverse events or complications in both groups. The median number of microemboli was 15 (range 5-48) in Group A and 68 (range 35-290) in Group B (P<0.05). CONCLUSION: The Enclose II device can be a valuable tool to perform venous or arterial proximal anastomoses during off-pump CABG. Transcranial Doppler results suggest that the device seems to be less traumatizing than side clamping and may reduce clamp-associated complications during off-pump coronary artery surgery. PMID- 15896620 TI - Usefulness of procalcitonin in the early detection of infection after thoracic surgery. AB - OBJECTIVE: The twofold aim of this prospective clinical study was to assess the accuracy of procalcitonin as a marker of postoperative infection after thoracic surgery and to compare it with C-reactive protein. METHODS: Procalcitonin and C reactive protein concentrations, clinical symptoms of infection and systemic inflammation were recorded preoperatively and 5 days postoperatively in 157 patients undergoing the following procedures: 52 wedge resections, 28 pneumonectomies and 77 lobectomies (or bilobectomies). Patients were classified as non-infected or infected according to predefined criteria. RESULTS: In non infected patients (n=132), procalcitonin peaked on day 1 and C-reactive protein, on day 2. The procalcitonin value was significantly higher in patients having undergone a pneumonectomy (0.73+/-0.78 versus 0.54+/-0.25 ng/mL for lobectomy and 0.50+/-0.35 ng/mL for wedge resection; P=0.04). The mean value of procalcitonin was significantly higher in patients with postoperative infection (n=25) than in those with no postoperative infection (3.6+/-5.5 versus 0.63+/-0.62 ng/mL; P=0.0001). The onset of infection most frequently occurred on postoperative day 2 (43% of patients); maximum procalcitonin and C-reactive protein concentrations most frequently appeared on postoperative day 1 (56% of patients) and day 2 (63% of patients), respectively. The best cutoff value for detection of infection with procalcitonin was 1 ng/mL and with C-reactive protein, 100mg/L. Comparing the area under the Receiver Operating Characteristic curves, procalcitonin was better than C-reactive protein for detecting postoperative infection (0.92 versus 0.66; P<0.0001). CONCLUSIONS: Procalcitonin can be used as a reliable diagnostic parameter to detect and to monitor infectious complications in the postoperative period after thoracic surgery, especially in patients felt to be at higher risk (SIRS). It provides more information about the course of the disease than C reactive protein does, and can be detected before the occurrence of clinical infection. PMID- 15896621 TI - Video-assisted transcatheter lung perfusion regional chemotherapy. AB - OBJECTIVE: To establish a technique for performing isolated lung perfusion (ILP) under video-assisted thoracic surgery (VATS) to treat unresectable lung malignancies. METHODS: Under fluoroscopic and thoracoscopic guidance, five canine left lungs were isolated by means of an endovascular technique comprising pulmonary artery cannulation through the right femoral vein and pulmonary vein cannulation through the left auricular appendage (VATS-ILP). ILP was performed for 20 min at a flow rate of 30 ml/min with a high-dose cisplatin solution (50 microg/ml). Toxicity and pharmacokinetics of VATS-ILP were compared with those of conventional ILP performed in five additional lungs. RESULTS: VATS-ILP was performed safely without adverse reaction. Both VATS-ILP and conventional ILP delivered a high dose of cisplatin to the treated lung (total platinum concentration: 48+/-17 microg/g tissue for VATS-ILP vs. 51+/-19 microg/g tissue for conventional ILP, P>0.1) without significant systemic leakage (total platinum concentration: 0.4+/-0.1 microg/ml plasma vs. 0.5+/-0.2 microg/ml plasma, P>0.1). In addition, no significant differences were observed between the groups in the serum lactate dehydrogenase level, serum angiotensin-converting enzyme level, body weight change, or mid-term histological change following ILP. A significantly smaller thoracotomy was used for VATS-ILP than for conventional ILP (4.7+/-0.4 cm for VATS-ILP vs. 12+/-0.7 cm for conventional ILP, P<0.001) because VATS-ILP required neither arteriotomy nor venotomy. CONCLUSIONS: We established a canine VATS-ILP model that showed pharmacokinetic potential similar to that of conventional ILP. A clinical trial of VATS-ILP with cytotoxic drugs is warranted. PMID- 15896622 TI - Anterograde versus retrograde isolated lung perfusion with melphalan in the WAG Rij rat. AB - OBJECTIVE: Isolated lung perfusion (ILuP) is an experimental technique currently tested to increase the 5-year survival of 40% after surgical resection of pulmonary metastases from certain solid tumors. The standard technique of anterograde perfusion was compared with retrograde isolated lung perfusion in which the drug is introduced through the pulmonary veins while the effluent is collected from the pulmonary artery. Since the lung has a dual arterial circulation through the pulmonary artery and bronchial circulation, perfusion through the pulmonary veins can result in a more homogeneous distribution throughout the lung with subsequent higher melphalan concentration. METHODS: We randomized 20 rats into two groups. Group one underwent anterograde isolated left lung perfusion while group two underwent retrograde isolated left lung perfusion. A dose of 2 mg/kg melphalan (MN) was administered to the lung at a flow of 0.5 mL/min during 30 min, followed by a 5-min washout with buffered hetastarch (BHE). The final melphalan lung concentration (FMLC) was determined in the hilum, at the apex, the mid-periphery and the base of the lung. Statistical analysis was done with an unpaired student's t-test. RESULTS: Retrograde left ILuP resulted in a higher FMLC in the hilum (P<0.0001) and in the base of the lung (P=0.03), while anterograde ILuP induced a higher concentration at the apex of the lung (P=0.04). No difference was seen in the mid-peripheral area of the lung (P=0.92). CONCLUSIONS: In this experimental study, retrograde perfusion seems to increase final melphalan lung concentration in hilar and basal regions of the lung compared to anterograde perfusion. PMID- 15896623 TI - Postoperative adjuvant chemotherapy for stage I non-small cell lung cancer. AB - OBJECTIVE: Surgery constitutes the mainstay of treatment in stage I non-small cell lung cancer (NSCLC). However, a significant fraction of patients after surgical resection die mainly due to systemic relapse. Nonetheless, the best adjuvant treatment to improve survival and decrease relapse rate remains as an ever controversial issue. Therefore, we conducted a randomized trial to determine whether postoperative adjuvant chemotherapy is beneficial in prolonging survival and decreasing recurrence in patients with completely resected stage I NSCLC. METHODS: It was designed as a randomized, prospective two-armed study with surgery only (control group, 59 patients) versus surgery plus adjuvant MVP (mitomycin C, vinblastin and cisplatin) chemotherapy (study group, 59 patients). RESULTS: Data for all the patients were complete. Twenty-four patients in the control group and nine patients in the study group experienced tumor recurrence during the follow-up. Neither histological type nor surgical extent correlated with recurrence. However, the addition of adjuvant MVP chemotherapy could decrease the rate of recurrence and the incidence of cancer-related death after surgery in the patients of stage I NSCLC (P<0.05). We followed up at least 5 years, and the duration of mean follow-up was 7.3 years. The rates of the loco regional and distant metastases were 3.4 and 40.7% in the control group, and 3.4 and 11.9% in the study group, respectively. The 5- and 10-year survival rates were 74.6 and 56.3% in the control group, and 81.4 and 65.0% in the study group, respectively (P=0.19, log-rank test). The 5- and 10-year disease-free survival rates were 64.8 and 54.8% in the control group, and 88.8 and 76.8% in the study group, respectively (P=0.002, log-rank test). CONCLUSIONS: Our results suggest that the addition of adjuvant MVP chemotherapy may reduce the incidence of distant metastasis and prolong the disease-free survival of the patients with stage I NSCLC after surgery. PMID- 15896624 TI - Comparison of neoadjuvant cisplatin-based chemotherapy versus radiochemotherapy followed by resection for stage III (N2) NSCLC. AB - OBJECTIVE: Comparison of prospectively treated patients with neoadjuvant cisplatin-based chemotherapy vs radiochemotherapy followed by resection for mediastinoscopically proven stage III N2 non-small cell lung cancer with respect to postoperative morbidity, pathological nodal downstaging, overall and disease free survival, and site of recurrence. METHODS: Eighty-two patients were enrolled between January 1994 to June 2003, 36 had cisplatin and doxetacel-based chemotherapy (group I) and 46 cisplatin-based radiochemotherapy up to 44 Gy (group II), either as sequential (25 patients) or concomitant (21 patients) treatment. All patients had evaluation of absence of distant metastases by bone scintigraphy, thoracoabdominal CT scan or PET scan, and brain MRI, and all underwent pre-induction mediastinoscopy, resection and mediastinal lymph node dissection by the same surgeon. RESULTS: Group I and II comprised T1/2 tumors in 47 and 28%, T3 tumors in 45 and 41%, and T4 tumors in 8 and 31% of the patients, respectively (P=0.03). There was a similar distribution of the extent of resection (lobectomy, sleeve lobectomy, left and right pneumonectomy) in both groups (P=0.9). Group I and II revealed a postoperative 90-d mortality of 3 and 4% (P=0.6), a R0-resection rate of 92 and 94% (P=0.9), and a pathological mediastinal downstaging in 61 and 78% of the patients (P<0.01), respectively. 5y overall survival and disease-free survival of all patients were 40 and 36%, respectively, without significant difference between T1-3 and T4 tumors. There was no significant difference in overall survival rate in either induction regimens, however, radiochemotherapy was associated with a longer disease-free survival than chemotherapy (P=0.04). There was no significant difference between concurrent vs sequential radiochemotherapy with respect to postoperative morbidity, resectability, pathological nodal downstaging, survival and disease free survival. CONCLUSIONS: Neoadjuvant cisplatin-based radiochemotherapy was associated with a similar postoperative mortality, an increased pathological nodal downstaging and a better disease-free survival as compared to cisplatin doxetacel-based chemotherapy in patients with stage III (N2) NSCLC although a higher number of T4 tumors were admitted to radiochemotherapy. PMID- 15896625 TI - Early and long-term results of lung resection for non-small-cell lung cancer in patients with severe ventilatory impairment. AB - OBJECTIVE: To study clinical characteristics, surgical treatment modalities, early and long-term outcome of patients with severe ventilatory impairment undergoing lung resection for NSCLC. METHODS: We performed a retrospective review of clinical records of all patients with severe chronic ventilatory impairment (FEV1 and/or FVC< or =50% of predicted values) operated on for NSCLC in a 21-year period (1983-2003). RESULTS: One hundred and six patients were operated on. Mean FEV1 and FVC were 40% (range 23-50%) and 69% (17-117%), respectively. An obstructive pattern was observed in 87 cases (82%). Extent of maximal exeresis was based on the assessment of predicted post-operative FEV1 (ppoFEV1). Major resections were contraindicated if ppoFEV1 was lower than 30%. Sixteen pneumonectomies, 73 lobectomies and 17 sublobar resections were carried out. Pathologic stages were I, II, IIIA and IIIB in 58, 26, 18 and 4 cases, respectively. Resection was complete in 104 patients. Operative mortality and morbidity were 8.5% (n=9) and 70% (n=74), respectively. Twenty-two patients needed prolonged (>48 h) mechanical ventilation. Overall mean ppoFEV1 loss was 9.1% (0-34%). If ppoFEV1 loss was >15%, the morbidity rate was 100%. Mean PaCO2 and ppoFEV1 loss were higher among patients who died (41 mmHg versus 37 mmHg, P=0.02 and 13.2% versus 8.5%, P=0.025, respectively) as compared with operative survivors. Among patients with PaCO2>39 mmHg and ppoFEV1 loss>15% (n=9), mortality rate was 33%. Overall 1-year and 5-year survival rates were 82 and 33%, respectively. Respiratory failure was the cause of late death in 2 patients. Among patients available at follow-up (n=85), respiratory function was considered subjectively improved, stable and worsened in 6 (7%), 62 (73%) and 17 (20%) cases, respectively. Eleven patients needed continuous oxygen therapy. CONCLUSIONS: Lung resection should not be denied a priori in patients with severe ventilatory impairment. Evaluation of predicted post-operative function often allows major resections, which are functionally economic, at the price of a high operative morbidity. Operative mortality, long-term survival and respiratory function are acceptable in the absence of a valid therapeutic alternative. PMID- 15896626 TI - Prognostic significance of microvascularization in cases of operated lung cancer. AB - OBJECTIVE: Measurement of microvascularization and determination of its prognostic significance in cases of lung cancer. METHODS: Section prepared from histological material from 432 radically operated non-small cell lung cancer patients were stained with antibodies against factor VIII-associated antigen. During computer imaging, the absolute and relative parameters of the vascularization were determined, as was the density of tumour cells situated in the vicinity of the vessels. The results were compared with the TNM status, the cell type and the survival. RESULTS: Each parameter demonstrated an enhanced vascularization in classifications T2 and T4, but only the surface fraction, the mean vascular circumference and the mean vascular area displayed a significant change. The microvascularization parameters did not differ significantly between with different N status, however, the cell density progressively increased in the areas close to the vessels in advanced pN classifications. Elevation of the tumour cell density within 20 microm distance of the vessels was accompanied by a significantly poorer survival rate. The density of tumour cells within 20 microm region was the second strongest prognostic factor after the N status. CONCLUSIONS: More advanced tumour classifications grow with enhanced vascularization. A clear-cut connection cannot be demonstrated between the vascularization and appearance of lymph node metastases. The density of tumour cells measured in the direct vicinity of vessels is an important prognostic factor. PMID- 15896627 TI - An unusual foreign body in the tracheobronchial tree. PMID- 15896628 TI - Intrathoracic rib: multidetector computed tomography demonstration. PMID- 15896629 TI - Aortic valve replacement concomitant with multiple extra-anatomical bypasses for a patient with aortic valve insufficiency having Takayasu's arteritis. PMID- 15896630 TI - Obstructive thrombosis of an aortic root homograft. PMID- 15896631 TI - Pacemaker dependent patients with device infection--a modified approach. AB - A modified surgical concept for temporary cardiac pacing in pacemaker dependent patients requiring total removal of infected devices is presented. Proximal to the infected pocket a permanent bipolar pacing lead is placed transcutaneously into the ipsilateral subclavian or jugular vein. The lead is placed in the right ventricle and fixed into the skin using the suture sleeve. Pacing is established by connecting an external pacing generator. Subsequently the infected device can be removed completely. After wound dressing the externalized lead is connected to a permanent VVI-pacemaker allowing for prolonged temporary pacing. PMID- 15896632 TI - Sternum lifting technique for thoracoscopic internal thoracic artery harvest. AB - We employed the Laparolift and Laparofan (Origin Medsystems Inc., CA, USA) and developed a sternum lifting technique to create a sufficient intra-pleural space between the heart and the sternum in which the left internal thoracic artery (ITA) in situ graft could be thoracoscopically mobilized. Between June and December 2004, this technique was applied to 12 consecutive patients (eight men, four women, aged 68.5+/-9.6 years) undergoing minimally invasive coronary artery bypass grafting via a left mini-thoracotomy. The Laparofan, connected to the arm of the lift machine (Laparolift), was introduced through a subxyphoidal entry and opened beneath the sternum. The sternum was elevated by about 5 cm until a sufficient working space was created under the sternum. With left hemipulmonary collapse, the left ITA was thoracoscopically taken down through the axillary ports. There was no procedural conversion to direct harvesting. The mean thoracoscopic harvesting time was 34.5+/-7.7 min. There was no mortality and no instrument-related morbidities. Patency of each ITA graft was angiographically confirmed. In conclusion, despite the limited experience, the present sternum elevation technique using the Laparolift system is a viable method for increasing the intra-pleural working space beneath the sternum during thoracoscopic ITA harvesting. PMID- 15896633 TI - Portaclamp in video-assisted minimally invasive cardiac surgery: surgical technique and preliminary clinical experience. AB - Video-assisted minimally invasive cardiac surgery (VAMICS) is currently performed with various indications. However, despite the increasing evidence of its effectiveness, new approaches have to be defined to simplify this procedure, minimize its potential complications and limit its costs, for a wider use in the surgical community. The limited access to the aorta is a key point in VAMICS and mandates specific clamping modalities with their own limitations, costs and drawbacks. The Portaclamp (Cardio Life Research SA, Louvain la Neuve, Belgium), a new autoguided extravascular aortic cross-clamping system, has been recently proposed to facilitate VAMICS. Herein, we describe the Portaclamp approach and report our indications and preliminary clinical experience so to define its role in VAMICS. PMID- 15896634 TI - Two-stage surgical and endovascular treatment of an aneurysmal aberrant right subclavian (Lusoria) artery. AB - We report a case of a 75-year-old male patient with an asymptomatic aneurysm of an aberrant right subclavian artery (ARSA). Timely elective therapy of this entity is indicated due to the high risk of rupture. Because of the patient's reduced physical state, we performed an interdisciplinary endovascular aortic stent-grafting for the exclusion of the ARSA aneurysm. However, the proximal landing zone for the thoracic stent had to be chosen proximal to the origin of the left subclavian artery. Thus, 2 days before the endovascular procedure, a left-sided carotido-subclavian bypass was accomplished. The peri- and postoperative course was uneventful. The technical results in our patient were excellent, no leakage was visible so far. PMID- 15896636 TI - Ventricular myocardial band and Ross operation. PMID- 15896638 TI - Prognoses from the logistic EuroSCORE are statistical estimates that require confidence intervals. PMID- 15896639 TI - Aspergillosis complicating intralobar sequestration. PMID- 15896640 TI - Does the shuttle walk test correlate with postoperative morbidity in lung surgery? PMID- 15896643 TI - Investigating population level trends in head injuries amongst child cyclists in the UK. AB - Case control studies suggest that cycle helmets offer their wearers protection from injury in the event of an accident. Nevertheless, encouragement and even compulsion of cycle helmet wearing has been controversial. This paper will re examine another potential source of evidence for the role of cycle helmets. Administrative datasets are attractive because of their availability, but require careful analysis. The results presented here are obtained from analysing such data with an appropriate form of generalised additive model. Whilst helmet wearing surveys in the UK suggest strongly divergent trends in wearing rates between male and female children, there is little evidence from "Hospital Episode Statistics" to indicate similarly divergent trends in terms of head injury. Conversely, it can be confirmed that head injuries are falling faster among cyclists than pedestrians. Although case control studies suggest cycle helmets are not effective in reducing overall injuries, it is worth noting an increase in the proportion of male child cyclists reported by the police as being killed or seriously injured in road collisions. It might be tempting to use these results to suggest that helmets are not effective in reducing head injury at the population level. Whilst the careful analysis of population level data presented here is clearly important, this paper will discuss the reasons why population and individual level analyses of cycle helmets might be different and consider some of the difficulties in assigning cause and effect with imperfect observational data. PMID- 15896644 TI - Ageing effects on the expression of cell defence genes after UVA irradiation in human male cutaneous fibroblasts using cDNA arrays. AB - Ageing is a multifactorial process in which reactive oxygen species (ROS) are thought to be implicated. ROS cause oxidative alterations on cell constituents, and damage accumulation can lead to mutations in DNA. Modulation of gene expression during ageing is now quite documented but results are often controversial and/or incomplete. As ultraviolet A is one of the exogenous factors involved in skin ageing, by the production of ROS, we further document the modifications in gene expression during ageing process and response to an oxidative stress. For this purpose, we used a cDNA macroarray containing 82 genes related to cell defence, essentially represented by antioxidant and DNA repair proteins. Ageing-associated gene expression was assessed in normal skin human fibroblasts from three age groups: children (n=4), adults (n=4) and olders (n=3), at the basal state and after a 5J/cm2 UVA irradiation. Analysis revealed that 22 genes were never detected, whereas certain were always expressed such as those related to antioxidant defence, extracellular matrix (ECM) regulator and XPC. Transcripts related to ECM, MMP1 and MMP3 were increased with age and after UVA irradiation, independently of age. It appeared that transcripts involved in the redox status control (TXN and APEX) decreased as a function of age, at the basal state and after irradiation, respectively. Most of transcripts involved in DNA repair were not detected but repression of POLD1 in the adult group and induction of XRCC5 and LIG4 were observed after UVA irradiation, as a function of age. In the basal state, the transcript of GAS1, regulator of cell cycle arrest in G1 phase was found to be decreased with age. HMOX1 increased after UVA irradiation. In conclusion, the decrease in expression of some antioxidant system, cell cycle control gene and extracellular matrix enzymes, particularly after UV exposure can explain the occurrence of photoaging. PMID- 15896645 TI - Effects of the residues on the excitation energies of protonated Schiff base of retinal (PSBR) in bR: a TD-DFT study. AB - Effects of the residues on the excitation energies of protonated Schiff base of retinal (PSBR) in bacteriorhodopsin have been investigated by means of time dependent density functional theory. The residues around PSBR are replaced by the point charges on atoms. The structures of PSBR and residues are referred from X ray data. The atomic charges on the each residue were calculated the B3LYP/6 311G(d,p) level. The excitation energy of PSBR perturbed by the point charges on atoms of each residue was calculated at the B3LYP/6-31G(d,p) level. A total of 23 residues and five water molecules around PSBR were considered in the calculations. The large spectral shifts were caused by the Asp212 and Asp85. The origin of the spectral shifts was discussed on the basis of theoretical results. PMID- 15896646 TI - UVA-induced oxidative damage and cytotoxicity depend on the mode of exposure. AB - The reciprocity rule (Bunsen-Roscoe law) states that a photochemical reaction is directly proportional to the total energy dose, irrespective of the dose distribution. In photomedicine the validity of this law is usually taken for granted, although the influence of radiation intensity and dose distribution are largely unknown. We have examined in a tissue culture model the effects of fractionated versus single dose exposure to UV from a metal halide source on survival, DNA synthesis, glutathione, and oxidative membrane damage. Exposure to fractionated UVA was followed by an increased rate of cell death compared to single dose exposure, when intervals between fractions where short (10-120 min). Longer intervals had the opposite effect. Corresponding results were obtained for DNA synthesis (BrdU incorporation). The increased cytotoxicity of dose fractionation with short intervals could not be abrogated by non-enzymatic antioxidants (astaxanthin, ascorbic acid, alpha-tocopherol). Fractionated irradiation with short intervals led to higher degree of depletion of glutathione (GSH) and to enhanced formation of thiobarbituric acid reactive substances (TBARS) in comparison to an identical single dose. Long intervals between fractions induced opposite effects. Taken together, these data indicate that immediately after UVA exposure cells are more sensitive to a further oxidative attack making repeated exposure with short intervals more cytotoxic than continuous single dose UVA. This might have implications also for responses to UVA in vivo and further studies will have to extend these findings to the situation in healthy and diseased human skin. PMID- 15896648 TI - Techniques for delivery and monitoring of TOOKAD (WST09)-mediated photodynamic therapy of the prostate: clinical experience and practicalities. AB - Photodynamic therapy of solid organs requires sufficient PDT dose throughout the target tissue while minimizing the dose to proximal normal structures. This requires treatment planning for position and power of the multiple delivery channels, complemented by on-line monitoring during treatment of light delivery, drug concentration and oxygen levels. We describe our experience in implementing this approach in Phase I/II clinical trials of the Pd-bacteriophephorbide photosensitizer TOOKAD (WST09)-mediated PDT of recurrent prostate cancer following radiation failure. We present several techniques for delivery and monitoring of photodynamic therapy, including beam splitters for light delivery to multiple delivery fibers, multi-channel light dosimetry devices for monitoring the fluence rate in the prostate and surrounding organs, methods of measuring the tissue optical properties in situ, and optical spectroscopy for monitoring drug pharmacokinetics of TOOKAD in whole blood samples and in situ in the prostate. Since TOOKAD is a vascular-targeted agent, the design and implementation of the techniques are different than for cellular-targeted agents. Further development of these delivery and monitoring techniques will permit full on-line monitoring of the treatment that will enable real-time, patient-specific and optimized delivery of PDT. PMID- 15896649 TI - The vascular response to photodynamic therapy with ATX-S10Na(II) in the normal rat colon. AB - The mechanism of tissue damage from photodynamic therapy (PDT) may be cellular, vascular or both, depending on the photosensitising agent and the treatment conditions. Well established photosensitisers like porfimer sodium have an optimum drug light interval of two days and may cause skin photosensitivity lasting several weeks. ATX-S10Na(II) is a new photosensitiser that remains largely in the vasculature after systemic administration and clears from the body within a few hours. The present study looks at the factors controlling the extent of PDT necrosis using ATX-S10Na(II) and correlates these with changes in the circulation after PDT. Normal Wistar rats were sensitised with ATX-S10Na(II), 2 mg/kg. At laparotomy, a laser fibre was positioned just touching the colonic mucosa and 50 J light at 670 nm delivered varying the drug light interval (0.5-24 h) and light delivery regime (100 mW continuous, 20 mW continuous or 100 mW in five fractions). Some animals were killed at three days to document the area of necrosis, others received fluorescein shortly prior to death (from a few minutes to three days after PDT) to outline the zone of PDT induced vascular shutdown. Maximum necrosis was seen with the shortest drug light interval (0.5 h), with no effect by 6 h. Fractionating the light or lowering the power did not increase the necrosis. The area of fluorescein exclusion increased over the first 2 h after PDT (in contrast to the re-perfusion seen with other photosensitisers) and correlated with the area of necrosis. PDT with ATX-S10Na(II) is most effective with a drug light interval of less than one hour. It induces irreversible vascular shutdown that extends after completion of light delivery and which is largely independent of the light delivery regime. PMID- 15896650 TI - Determination of the distribution of light, optical properties, drug concentration, and tissue oxygenation in-vivo in human prostate during motexafin lutetium-mediated photodynamic therapy. AB - It is desirable to quantify the distribution of the light fluence rate, the optical properties, the drug concentration, and the tissue oxygenation for photodynamic therapy (PDT) of prostate cancer. We have developed an integrated system to determine these quantities before and after PDT treatment using motorized probes. The optical properties (absorption (micro(a)), transport scattering (micro(s'), and effective attenuation (micro(eff)) coefficients) of cancerous human prostate were measured in-vivo using interstitial isotropic detectors. Measurements were made at 732 nm before and after motexafin lutetium (MLu) mediated PDT at different locations along each catheter. The light fluence rate distribution was also measured along the catheters during PDT. Diffuse absorption spectroscopy measurement using a white light source allows extrapolation of the distribution of oxygen saturation StO2, total blood volume ([Hb]t), and MLu concentration. The distribution of drug concentration was also studied using fluorescence from a single optical fiber, and was found to be in good agreement with the values determined by absorption spectroscopy. This study shows significant inter- and intra-prostatic variations in the tissue optical properties and MLu drug distribution, suggesting that a real-time dosimetry measurement and feedback system for monitoring these values during treatment should be considered in future PDT studies. PMID- 15896651 TI - Monitoring PDT by means of superficial reflectance spectroscopy. AB - Monitoring of relevant parameters during photodynamic therapy (PDT) and correlating these with treatment response is necessary to guarantee optimal and reproducible treatment outcome. In this paper we study the correlation between changes in the local tissue optical properties (absorption and scattering coefficients) during ALA-PDT and changes in PpIX fluorescence. The optical properties are measured extremely superficially by employing a single fiber for the delivery and collection of white light to and from the tissue. The measured reflectance spectrum is modeled in terms of four relevant parameters: blood saturation, relative blood volume fraction, scattering intensity and wavelength dependence of the scattering. All these parameters, except the relative blood volume fraction, are shown to correlate with the rate of photobleaching of PpIX, which in turn has previously been shown to correlate with the response of tissues to PDT. These results yield valuable insight in the behavior of these parameters during PDT and their suitability to predict PDT-response for other photosensitizers for which monitoring through photobleaching is not possible. PMID- 15896652 TI - In vitro correction of medium chain acyl CoA dehydrogenase deficiency with a recombinant adenoviral vector. AB - Defects of mitochondrial beta-oxidation are a growing group of disorders with variable clinical presentations ranging from mild hypotonia to sudden infant death. Current therapy involves avoidance of fasting, dietary restrictions, and cofactor supplementation. Unfortunately, times of acute illness and noncompliance can interfere with these therapies and result in a rapid clinical decline. The development of a safe, durable, and effective gene delivery system remains an attractive alternative therapy for individuals with these disorders. To this end, a recombinant first-generation adenovirus vector (Ad/cmv-hMCAD) has been prepared that constitutively expresses the human medium chain acyl CoA dehydrogenase (MCAD) protein under the control of the CMV promoter and bovine polyadenylation signal. Characterization of human fibroblasts deficient in MCAD infected with Ad/cmv-hMCAD including Western analysis, immunohistological staining visualized with confocal microscopy, electron transfer protein (ETF) reduction assay, and palmitate loading studies was performed. Infection of MCAD deficient fibroblast with Ad/cmv-hmcad resulted in the production of a 55kDa protein that co-localized in cells with a mitochondrial marker. Extracts prepared from Ad/cmv-hMCAD infected deficient fibroblasts demonstrated correction of the block seen in the MCAD catalyzed reduction of ETF in the presence of octanoyl CoA. Finally, MCAD deficient fibroblasts infected with increasing amounts of Ad/cmv-hMCAD showed a stepwise improvement of the abnormal acylcarnitine profile exhibited by the deficient cells. Together these studies demonstrate our ability to express and monitor the expression of MCAD in treated cells and support further in vivo murine studies to assess toxicity and duration of correction with this and other MCAD recombinant vectors. PMID- 15896653 TI - Residual cholesterol synthesis and simvastatin induction of cholesterol synthesis in Smith-Lemli-Opitz syndrome fibroblasts. AB - Smith-Lemli-Opitz syndrome (RSH/SLOS) is an autosomal recessive, malformation syndrome caused by mutations in the 3beta-hydroxysterol delta7-reductase gene (DHCR7). DHCR7 catalyzes the reduction of 7-dehydrocholesterol (7DHC) to cholesterol. We report the mutation analysis and determination of residual cholesterol synthesis in 47 SLOS patients, and the effects of treatment of SLOS skin fibroblasts with simvastatin. Using deuterium labeling we have quantified the amount of synthesized cholesterol and 7DHC in homozygote, heterozygote, and control fibroblast cell lines. In SLOS fibroblasts, the fraction of synthesized cholesterol to total sterol synthesis ranged from undetectable to over 50%. This establishes that different mutant alleles encode enzymes with varying degrees of residual activity. There was a correlation between increased phenotypic severity and decreased residual cholesterol synthesis (r(2)=0.45, p<0.0001). Simvastatin treatment of SLOS fibroblasts with residual DHCR7 enzymatic activity decreased 7DHC levels and increased cholesterol synthesis. This increase in cholesterol synthesis is due to increased expression of a mutant allele with residual function. Determination of residual enzymatic activity for specific DHCR7 mutant alleles will help in understanding the processes underlying the broad phenotypic spectrum found in this disorder and will be useful in identifying patients who may benefit from simvastatin therapy. PMID- 15896654 TI - Neonatal screening for defects of the mitochondrial trifunctional protein. AB - Long-chain l-3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency has been included in the routine neonatal screening program by the German screening commission. As tandem mass spectrometry (TMS) does not discriminate between the different defects of the mitochondrial trifunctional protein (MTP) screening for isolated LCHAD deficiency includes the detection of long-chain 3-ketoacyl-CoA thiolase and complete MTP deficiencies as well. We identified 11 patients with abnormalities of the MTP out of 1.2 million newborns screened in our laboratory during the last 6 years. Treatment was started on the day the screening result was obtained (day 3 to day 9 of life). Seven of these newborns developed satisfactorily during an observation period of up to 64 months. They had isolated LCHAD deficiency, four of them caused by the typical mutation (1528 G>C), three others had no molecular genetic analysis done or were shown to have previously unknown mutations. Four children did not survive, two of them showing complete deficiency of MTP and two showing deficiency of long-chain 3-ketoacyl-CoA thiolase. We conclude that, despite the rarity of the disease, screening for MTP deficiencies is justified based on the following criteria: improved quality of life for patients with isolated LCHAD deficiency, absence of stigmatisation of babies showing mild variants without necessity of treatment, no significant increase of the total number of false positive screening results, no false negative results to our knowledge. Finally, extension of analysis to MTP deficiencies is achieved without additional costs for screening laboratories already using TMS. PMID- 15896655 TI - Prenatal diagnosis for severe methylenetetrahydrofolate reductase deficiency by linkage analysis and enzymatic assay. AB - Severe methylenetetrahydrofolate reductase (MTHFR) deficiency is characterized by varying degrees of developmental delay, motor and gait abnormalities, seizures, and thrombosis. Biochemical abnormalities include homocystinuria and hyperhomocysteinemia. Clinical severity correlates with MTHFR activity in cultured fibroblasts; activity can also be assayed in cultured amniocytes and chorionic villus cells (CVC). Forty-four private mutations have been identified, limiting the use of direct mutation detection for prenatal diagnosis. However, intragenic polymorphisms have been identified, making prenatal diagnosis by linkage analysis a possible option, even without knowledge of deleterious mutations. Prenatal diagnosis for severe MTHFR deficiency has been available by biochemical methodologies, but molecular genetic approaches have not yet been reported. We performed prenatal diagnosis for severe MTHFR deficiency in 11 at risk pregnancies in seven families. A combined approach of linkage analysis and enzymatic assays was used in six pregnancies; linkage analysis alone was performed in one pregnancy. Linkage analysis for the 677C > T or 1298A > C polymorphisms predicted that all seven fetuses were unaffected. For six of these seven fetuses, enzymatic activities were also measured and demonstrated concordant results. Of the 10 pregnancies in which enzymatic assays were performed, activities in cultured amniocytes predicted six unaffected fetuses (1.4-7.1 nmol CHO/mg prot/h (U)) and one affected fetus (0.24 U [control 3.1-9.6 U]). Three pregnancies assessed via CVCs demonstrated two unaffected fetuses (3.6 and 7.7 U) and 1 affected fetus (0 U [control 4.5-7.8 U]). These values were compared to those of the probands (range = 0.02-0.7 U (control 2.4-11.7 U)) in cultured fibroblasts. Our findings suggest that linkage analysis for severe MTHFR deficiency can be a practical approach for prenatal diagnosis. PMID- 15896656 TI - dif-1 and colt, both implicated in early embryonic development, encode carnitine acylcarnitine translocase. AB - It has always been assumed that during development the embryo and fetus depend only on glycolysis for energy generation and that they do not oxidize fatty acids. Recently, however, we found abundant expression and activity of fatty acid oxidation (FAO) enzymes in the human embryo and fetus. In a search for FAO gene expression during development we came across two embryonic differentiation genes: differentiation defective (dif-1) and congested-like trachea (colt) of Caenorhabditis elegans and Drosophila melanogaster, respectively. Earlier studies showed that expression of these two genes is essential during developmental stages with high energy requirements. Both dif-1 and colt encode proteins with sequence similarity to the mitochondrial carnitine acylcarnitine carrier (CACT), which suggests that the DIF-1 and COLT proteins might be functional orthologues of CACT. To investigate this, we expressed both dif-1 and colt in Saccharomyces cerevisiae. Our results show that DIF-1 and COLT can functionally complement a yeast CACT deletion strain and thus function as carnitine acylcarnitine transporters. This finding is well in line with the recent observation that embryos are capable of oxidizing fatty acids and furthermore implies that FAO is essential during early embryonic development when the energy demand is high. PMID- 15896657 TI - Two novel CHS1 (LYST) mutations: clinical correlations in an infant with Chediak Higashi syndrome. AB - Chediak-Higashi syndrome (CHS) is a rare autosomal recessive disease characterized by variable degrees of oculocutaneous albinism, recurrent infections, and a mild bleeding tendency, with late neurologic dysfunction. Most patients also undergo an accelerated phase of lymphohistiocytosis and die at an early age unless they receive an allogeneic hematopoietic stem cell transplant (SCT). Mutations in the CHS1 (LYST) gene result in CHS. Here, we describe an adopted infant who is compound heterozygous for two novel CHS1 gene mutations, both of which are predicted to result in truncated proteins. The two mutations are a nonsense mutation (c.1540 C>T, CGA>TGA, R514X) in exon 5 and a one base pair deletion (del c.9893T, F3298fsX3304) in exon 43, coding for part of the CHS1 protein's BEACH domain. These two newly described mutations are expected to give rise to a severe phenotype and, indeed, the patient had absolutely no cytotoxicity by natural killer cells or cytotoxic lymphocytes prior to his allogeneic SCT. PMID- 15896658 TI - Vesicle-associated membrane protein 4, a positional candidate gene on 1q24-q25, is not associated with type 2 diabetes in the Old Order Amish. AB - OBJECTIVE: The vesicle-associated membrane protein-4 (VAMP4) gene is an excellent type 2 diabetes (T2DM) positional candidate gene. It is located on chromosome 1q24-q25, a region of linkage to T2DM in the Amish and several other populations. VAMP4 is expressed in liver and skeletal muscle and participates in intracellular trafficking of secreted and membrane-associated proteins. DESIGN AND METHODS: We sequenced VAMP4 in 20 Amish subjects. Polymorphisms in and around VAMP4 were genotyped in 65 Amish subjects with T2DM, 64 subjects with impaired glucose homeostasis (IGH), and 126 normal glucose tolerant controls, as well as in an expanded set of 749 participants of the Amish Family Diabetes Study for whom glucose and insulin levels during an oral glucose tolerance test (OGTT) and other quantitative traits related to diabetes were available. Case-control and quantitative trait association analyses were performed. RESULTS: We found three common non-coding intragenic polymorphisms: a 23bp insertion/deletion (I/D) in the 5' untranslated region (UTR) in exon 1 at position 73127, and G35319T and C335296T single nucleotide polymorphisms (SNPs) in the 3' UTR (NCBI Accession No. Z98751). The two 3' UTR SNPs were in complete linkage disequilibrium (LD) and both were in strong LD with the exon 1 I/D polymorphism (|D'|=0.82). Similarly, three extragenic flanking SNPs (rs978985, rs203255, and rs1023479) showed moderate LD with the neighboring intragenic SNPs (|D'|=0.23-0.69). None of the SNPs individually nor any of the 2-, 3-, 4-, or 5-polymorphism haplotypes were associated with T2DM or IGH. The exon 1 I/D polymorphism was not associated with significant differences in mean fasting or stimulated glucose or insulin levels during an OGTT or other diabetes-related quantitative traits in the expanded set of 749 subjects. CONCLUSION: Variation in VAMP4 does not significantly influence risk of T2DM or IGH in the Amish. PMID- 15896659 TI - Study of a new PPARgamma2 promoter polymorphism and haplotype analysis in a French population. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) plays a role in adipocyte differentiation and insulin sensitization. We identified and characterized a new C/T substitution at position -689 (-689C>T) in the P2 promoter of PPARgamma in a putative GATA binding site. By electrophoretic mobility shift assay, both GATA2 and GATA3 proteins could bind weakly to the wild type P2 -689 GATA binding site but not to the mutated site. Neither GATA2 nor GATA3 was able to regulate significantly the P2 promoter activity in a reporter luciferase assay, whatever the allele at position -689 was, suggesting that the 689 putative GATA site was probably not a functional target for GATAs. However, the presence of the -689T allele rendered the P2 promoter less active at the basal state. We genotyped a population of 1155 men and women for the -689C>T polymorphism and looked for possible associations with anthropometric and lipid variables. The carriers of the -689T allele had elevated body weight and LDL cholesterol concentrations compared with the homozygous for the common allele. Haplotype analyses including the -681C>G (P3 promoter), -689C>T (P2 promoter), and Pro12Ala (exon B) polymorphisms were performed. Carriers of the G-T-Ala haplotype (corresponding to the P3 -681C>G, P2 -689C>T and Pro12Ala polymorphisms in this order) had elevated LDL-cholesterol concentrations and body weight compared with C-C-Pro individuals. In conclusion, we identified a new polymorphism in the P2 promoter of PPARgamma. The P3 -681C>G, P2 -689C>T, and Pro12Ala polymorphisms and related haplotypes were associated with higher body weight and plasma LDL-cholesterol concentrations. PMID- 15896660 TI - The -374A allele of the receptor for advanced glycation end products gene is associated with a decreased risk of ischemic heart disease in African-Brazilians with type 2 diabetes. AB - Three functional polymorphisms described in the promoter of receptor for advanced glycation end products (RAGE) gene were shown to have a marked effect on transcriptional activity. The few studies which analyzed the relationship between these three polymorphisms and the diabetic complications have shown conflicting results. In this case-control study, we evaluated the association between the 429T>C, the -374T>A and the 63bp insertion/deletion (I/D) polymorphisms in the RAGE gene, and the presence of diabetic retinopathy, diabetic nephropathy and ischemic heart disease, in 703 Brazilians with type 2 diabetes (520 Caucasian- and 183 African-Brazilians). Patients underwent a clinical and laboratory evaluation consisting of a questionnaire, physical examination, assessment of diabetic complications and blood collection. Genotype analysis was performed using the polymerase chain reaction and allele-specific restriction. Logistic regression analyses were used to examine associations between the clinical and genetic variables and the presence of diabetic complications. No association between the -429C, the -374A and the 63bp D alleles and diabetic retinopathy, diabetic nephropathy or ischemic heart disease was observed in Caucasian Brazilians with type 2 diabetes. However, the -374A allele was associated with a decreased risk of having ischemic heart disease in African-Brazilian type 2 diabetic patients [odds ratio (OR)=0.35; 95% confidence interval (CI)=0.15-0.81; P=0.014], independently of other risk factors associated with this complication. Thus, our results show that the -374A allele (-374T>A polymorphism) in the RAGE gene is related to the susceptibility of developing ischemic heart disease in African-Brazilians with type 2 diabetes. PMID- 15896661 TI - Reduced incidence of severe metabolic crisis or death in children with medium chain acyl-CoA dehydrogenase deficiency homozygous for c.985A>G identified by neonatal screening. AB - The incidence of severe metabolic crises in medium chain acyl-CoA dehydrogenase deficiency (MCADD) patients homozygous for the common c.985A>G mutation, who had been identified by neonatal screening, was assessed prospectively and compared to retrospective cohort data in unscreened patients with identical genotypes. Logrank test showed a significant reduction of severe metabolic crises in the screened cohort (p<0.01). Neonatal screening appears to reduce the rate of severe metabolic crisis or death in the most prevalent subset of MCADD. PMID- 15896662 TI - Biochemical and genetic characterization of four cases of hereditary coproporphyria in Spain. AB - We report a biochemical and genetic characterization of four cases of hereditary coproporphyria (HCP) in Spain. All patients showed a typical HCP porphyrin excretion pattern with a high concentration of coproporphyrins in feces and inverted I:III isomer ratio. The porphyrin precursors in urine were found elevated in two patients who showed acute symptoms. The analysis of the CPO gene showed that three cases harboured novel mutations: V135A (404T>C; exon 1); L214R (641T>G; exon 2); and P249R (746C>G; exon 3) and in the fourth, a previously described R426X mutation in exon 6. PMID- 15896663 TI - The sunburn cell: regulation of death and survival of the keratinocyte. AB - Sunburn cells are keratinocytes undergoing apoptosis after they have received a physiological UVB dose that irreversibly and severely damaged their DNA or other chromophores. If these cells would escape programmed cell death, a cancer prone phenotype could arise. On the other hand, if the decision to die is made too prematurely, the proliferative compartment of basal keratinocytes would be inevitably lost, thereby hampering normal skin homeostasis. Pro- and anti apoptotic mediators carefully control crucial points of the cell death program by regulating complex signalling cascades originating at the cell membrane, the nucleus and the cytoplasm. The balance between survival and apoptogenic factors determines the final cell fate, and growing evidence suggests that the deregulation of this balance by chronic UVB stress, results in the development of skin malignancy. The present paper reviews recent data on the major pathways regulating UVB-induced sunburn cell formation and implicates the deregulation of these pathways in the development of skin cancer. PMID- 15896664 TI - Thick ascending tubular cells in the loop of Henle: regulation of electrolyte homeostasis. AB - Renal medullary tubular cells in the loop of Henle have crucial importance for the regulation of homeostasis of the extracellular fluid. These cells receive limited amount of blood and oxygen, and are also constantly challenged by the hypertonic environment. The medullary tubular cells in the last part of the loop of Henle have one of the highest known contents of mitochondria of all mammalian cells, reflecting their need for oxidative metabolism in order to sustain high ATP production for active transepithelial electrolyte transport. The commonly used diureticum furosemide targets one of the transporters present in these tubular cells with resulting diuresis. Several pathological states are associated with altered function of the medullary tubular cells, and the nephrotoxic substances tacrolimus and cyclosporine act on these cells. The specific Tamm Horsfall glycoprotein is produced by medullary tubular cells. Alteration in the urinary excretion of this protein is used as marker of tubular damage. PMID- 15896665 TI - Ebola virus: the role of macrophages and dendritic cells in the pathogenesis of Ebola hemorrhagic fever. AB - Ebola hemorrhagic fever is a severe viral infection characterized by fever, shock and coagulation defects. Recent studies in macaques show that major features of illness are caused by effects of viral replication on macrophages and dendritic cells. Infected macrophages produce proinflammatory cytokines, chemokines and tissue factor, attracting additional target cells and inducing vasodilatation, increased vascular permeability and disseminated intravascular coagulation. However, they cannot restrict viral replication, possibly because of suppression of interferon responses. Infected dendritic cells also secrete proinflammatory mediators, but cannot initiate antigen-specific responses. In consequence, virus disseminates to these and other cell types throughout the body, causing multifocal necrosis and a syndrome resembling septic shock. Massive "bystander" apoptosis of natural killer and T cells further impairs immunity. These findings suggest that modifying host responses would be an effective therapeutic strategy, and treatment of infected macaques with a tissue-factor inhibitor reduced both inflammation and viral replication and improved survival. PMID- 15896666 TI - The roles of interleukin-15 receptor alpha: trans-presentation, receptor component, or both? AB - Interleukin-15 receptor alpha (IL-15R alpha) is a high affinity IL-15 binding protein that is crucial for mediating IL-15 functions such as memory CD8 T cell proliferation and NK, NK/T cell, and intestinal intraepithelial lymphocyte development. However, the mechanism by which IL-15R alpha mediates IL-15 functions is unique among cytokines. Originally, IL-15R alpha was thought to be a component of a heterotrimeric receptor complex containing the IL-2/IL-15R beta and common gamma chains (gammaC) that were required for mediating signaling. Although IL-15R alpha may in some cases act as a component of this receptor complex, more recent evidence indicates that IL-15R alpha predominately functions by presenting IL-15 to opposing cells expressing the IL-15R betagamma signaling components. This theory is consistent with the broad, non-lymphoid expression pattern of IL-15R alpha and the evidence that IL-15R alpha expression by lymphocytes is dispensable for IL-15 action in vivo. This new concept of cytokine delivery will allow us to better understand the regulation and function IL-15. PMID- 15896667 TI - Aurora kinases. AB - Aurora kinases A (also known as Aurora, Aurora-2, AIK, AIR-1, AIRK1, AYK1, BTAK, Eg2, MmIAK1 and STK15), Aurora B (also known as Aurora-1, AIM-1, AIK2, AIR-2, AIRK-2, ARK2, IAL-1 and STK12) and Aurora C (also known as AIK3) participate in several biological processes, including cytokinesis and dysregulated chromosome segregation. These important regulators of mitosis are over-expressed in diverse solid tumors. One member of this family of serine-threonine kinases, human Aurora A, has been proposed as a drugable target in pancreatic cancer. The recent determination of the three-dimensional structure of Aurora A has shown that Aurora kinases exhibit unique conformations around the activation loop region. This property has boosted the search and development of inhibitors of Aurora kinases, which might also function as novel antioncogenic agents. PMID- 15896668 TI - Effects of repeated administration with CP-55,940, a cannabinoid CB1 receptor agonist on the metabolism of the hepatic heme. AB - Drugs metabolised by cytochrome P450 (CYP) such as analgesics may induce acute attacks in patients with hepatic porphyrias. In recent years, preclinical and clinical studies have suggested that cannabinoid pharmaceutical preparations may be potentially useful in the treatment of pain. The purpose of the study was to examine the effects of CP-55,940, a cannabinoid CB1 receptor agonist, on the hepatic heme metabolism in mice. To this end, hepatic activities of aminolevulinic acid synthase (ALAS), heme oxygenase (HO) and CYP levels were determined in mice treated with CP-55,940 (0.5 mg/kg/day; i.p.; 5 or 24 days). Results showed that treatment with CP-55,940 decreased CYP concentrations by 80% and increased HO activity by 158%. However, ALAS activity also decreased by 37%, suggesting that regulatory free heme pool was not modified. Our findings indicate that CP-55,940 and its metabolites do not behave as porphyrinogenic drugs and may potentially be safe for treating pain in patients with acute porphyrias. PMID- 15896669 TI - Vulgarinin, a broad-spectrum antifungal peptide from haricot beans (Phaseolus vulgaris). AB - From the seeds of haricot beans (Phaseolus vulgaris), an antifungal peptide with a molecular mass around 7 kDa was purified by using a simple protocol consisting of affinity chromatography on Affi-gel blue gel and gel filtration on Superdex 75. This peptide named vulgarinin manifested an antifungal activity toward fungal species such as Fusarium oxysporum, Mycosphaerella arachidicola, Physalospora piricola and Botrytis cinerea, and an antibacterial action on Mycobacterium phlei, Bacillus megaterium, Bacillus subtilis and Proteus vulgaris. It also inhibited proliferation in leukemia cell lines L1210 and M1 and breast cancer cell line MCF-7. This peptide could reduce the activity of HIV-1 reverse transcriptase and inhibited translation in a cell-free rabbit reticulocyte lysate system. Its antifungal activity was retained after incubation with trypsin. PMID- 15896670 TI - CKLFSF2 is highly expressed in testis and can be secreted into the seminiferous tubules. AB - CKLFSF2 is a member of the chemokine-like factor superfamily (CKLFSF), a novel gene family containing CKLF and CKLFSF1-8. Using a combination of data mining and polymerase chain reactions, we determined the full cDNA sequence and genomic structure of human CKLFSF2, a 4-exon gene encoding 248 amino acids and spanning approximately 8.8 kb on chromosome 16q22.1. Expression profile analyses indicated that CKLFSF2 is expressed in a limited number of tissues. Specifically, immunohistochemistry indicated that CKLFSF2 is highly expressed in testis, mainly in spermatogonia and the seminiferous tubular fluid. Subcellular localization experiments suggested that CKLFSF2 is equally distributed in the cytoplasm, and Western blot analysis revealed that overexpressed CKLFSF2 is secreted into the supernatant of cultured cells. The data therefore strongly suggest that CKLFSF2 is a secreted protein that may be functionally relevant during spermatogenesis. PMID- 15896671 TI - Chromosome 1 open reading frame 10 (C1orf10) gene is frequently down-regulated and inhibits cell proliferation in oral squamous cell carcinoma. AB - Chromosome 1 open reading frame 10 (C1orf10) is a recently identified gene encoding a protein with an S100 EF-hand calcium-binding motif, and its expression is known to be down-regulated in esophageal squamous cell carcinoma. In this study, to determine whether the loss of C1orf10 gene function could contribute to the development of oral squamous cell carcinoma (OSCC), we have evaluated the expression status of this gene by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and quantitative real-time PCR analysis. A high frequency of decrease in C1orf10 gene was detected not only in OSCC-derived cell lines but also in tumor tissues. Next, to define biological function of this gene in oral carcinogenesis, we transfected Clorf10 with an Ecdysone-inducible system in OSCC cell lines and analyzed the effects of its overexpression. Induction of C1orf10 expression resulted in a significant decline in the rate of cell proliferation, and in an arrest in the G1 phase of the cell cycle, with a down-regulation of Cyclin D1 expression. However, we could not detect significant difference in the percentage of apoptotic cells. Thus, our results suggest that the down-regulation of C1orf10 gene plays a role in oral carcinogenesis, and that its expression may negatively regulate OSCC cell proliferation by arresting the cell cycle. PMID- 15896672 TI - The influence of phospholipid membranes on bovine calcitonin peptide's secondary structure and induced neurotoxic effects. AB - The peptide hormone, calcitonin, which is associated with medullary carcinoma of the thyroid, has a marked tendency to form amyloid fibrils and may be a useful model in probing the role of peptide-membrane interactions in beta-sheet and amyloid formation and amyloid neurotoxicity. Using bovine calcitonin, we found that, like other amyloids, the peptide was toxic only when in a beta-sheet-rich, amyloid form, but was non-toxic, when it lacked an amyloid structure. We found that the peptide bound with significant affinity to membranes that contained either cholesterol and gangliosides. In addition, incubation of calcitonin with cholesterol-rich and ganglioside-containing membranes resulted in significant changes in peptide structure yielding a peptide enriched in beta-sheet and amyloid content. Because the cholesterol- and ganglioside-rich phospholipid systems enhanced the calcitonin beta-sheet and amyloid contents, and peptide amyloid content was associated with neurotoxicity, we then investigated whether depleting cellular cholesterol and gangliosides affected calcitonin neurotoxicity. We found that cholesterol and ganglioside removal significantly reduced the calcitonin-induced PC12 cell neurotoxicity. Similar results have been observed with other amyloid-forming peptides such as beta-amyloid (A beta) of Alzheimer's disease and suggest that modulation of membrane composition and peptide-membrane interactions may prove useful in the control of amyloid formation and amyloid neurotoxicity. PMID- 15896673 TI - Hyperoxia results in transient oxidative stress and an adaptive response by antioxidant enzymes in goldfish tissues. AB - The effects of hyperoxia on the status of antioxidant defenses and markers of oxidative damage were evaluated in goldfish tissues. The levels of lipid peroxides, thiobarbituric acid reactive substances, carbonyl proteins and the activities of some antioxidant enzymes were measured in brain, liver, kidney and skeletal muscle of goldfish, Carassius auratus L., over a time course of 3-12 h of hyperoxia exposure followed by 12 or 36 h of normoxic recovery. Exposure to high oxygen resulted in an accumulation of protein carbonyls in tissues throughout hyperoxia and recovery whereas lipid peroxides and thiobarbituric acid reactive substances accumulated transiently under short-term hyperoxia stress (3 6 h) but were then strongly reduced. This suggests that hyperoxia stimulated an enhancement of defenses against lipid peroxidation or mechanisms for enhancing the catabolism of peroxidation products. The activities of principal antioxidant enzymes, superoxide dismutase and catalase, were not altered under hyperoxia but catalase increased during normoxic recovery; activities may rise in anticipation of further hyperoxic excursions. In most tissues, the activities of glutathione utilizing enzymes (glutathione peroxidase, glutathione-S-transferase, glutathione reductase) as well as glucose-6-phosphate dehydrogenase, were not affected under hyperoxia but increased sharply during normoxic recovery. Correlations between some enzyme activities and oxidative stress markers were found, for example, an inverse correlation was seen between levels of thiobarbituric acid reactive substances and glutathione-S-transferase activity in liver and catalase and glucose-6-phosphate dehydrogenase in kidney. The results suggest that liver glutathione-S-transferase plays an important role in detoxifying end products of lipid peroxidation accumulated under hyperoxia stress. PMID- 15896674 TI - Novel ryanodine-binding properties in mammalian retina. AB - The ryanodine receptor (RyR)/Ca2+ release channel mobilizes Ca2+ from internal calcium stores to support a variety of neuronal functions. To investigate the presence of such a protein in mammalian retina, we applied ryanodine binding, PCR and antibodies against known RyRs. Surprisingly, ryanodine-binding properties of retinal endoplasmic reticulum-enriched membrane fraction were vastly different from those of skeletal and cardiac muscles ryanodine-binding proteins. In common with the skeletal and cardiac muscle, ryanodine bound with high-affinity to two or more types of binding site (Kd1 = 20.6 and Kd2 = 114 nM); binding was strongly stimulated by high concentrations of NaCl; it was inhibited by tetracaine and the protein appeared to possess an ATP-binding site. Unlike cardiac and skeletal muscle, RyRs in retina binding was Ca2+-independent; inhibited by caffeine and dantrolene; less sensitive to ruthenium red; and unaffected by La3+. Also, in retina, ryanodine rapidly associated to and dissociated from its binding sites. Furthermore, although the protein bound the ATP analog BzATP, retinal ryanodine binding was not stimulated by nucleotides. Immunostaining of bovine retinal sections with anti-RyR2 showed a strong staining of amacrine, horizontal and ganglion cells. Finally, using RT-PCR, the three known RyR isoforms were identified in retina. However, consistent with the novel binding properties, the peptide maps yielded by trypsin treatment and Western blotting demonstrate different patterns. Together, the results suggest that retina expresses a novel ryanodine-binding protein, likely to be a ryanodine receptor. Its presence in retina suggests that this protein might play a role in controlling intracellular Ca2+ concentration. PMID- 15896675 TI - How can data quality and automation enhance confidence in microarray data? PMID- 15896676 TI - Canadian drug reimportation debate goes on. PMID- 15896677 TI - Getting inside the bug: new targets for TB drugs. PMID- 15896678 TI - Looking for new sepsis targets. PMID- 15896679 TI - Jerome Skelly on regulation, education and information. PMID- 15896680 TI - A viewpoint on South Korean Biotech. PMID- 15896681 TI - Sourcing a chemical succession for cyclosporin from parasites and human pathogens. PMID- 15896682 TI - Keynote review: Medicinal chemistry strategies to CB1 cannabinoid receptor antagonists. AB - The proven clinical efficacy of the CB(1) cannabinoid receptor antagonist rimonabant in both obesity and smoking cessation and its therapeutic potential in other disorders has given a tremendous impetus to the discovery of novel CB(1) antagonists. The number of disclosed patents wherein novel chemical entities having CB(1) antagonistic or inverse agonistic properties have been claimed has exploded. Besides novel compound classes that were identified in screening, rational medicinal chemistry approaches such as conformational constraint and scaffold hopping have been successfully applied. CB(1) receptor modelling has provided insight into crucial receptor-ligand interaction points thereby leading to a general CB(1) inverse agonist pharmacophore model. PMID- 15896683 TI - Incretin mimetics and DPP-IV inhibitors for the treatment of type 2 diabetes. AB - Incretin mimetics are a new class of pharmacological agents with multiple antihyperglycemic actions that mimic the actions of incretin hormones such as glucagon-like peptide (GLP)-1. Dipeptidyl peptidase (DPP-IV) inhibitors suppress the degradation of many peptides, including GLP-1, thereby extending their bioactivity. Several incretin mimetics and DPP-IV inhibitors are undergoing late stage clinical trials for the treatment of type 2 diabetes. These agents appear to have multiple mechanisms of action, including some or all of the following: enhancement of glucose-dependent insulin secretion; suppression of inappropriately elevated glucagon secretion; slowing of gastric emptying; and decreased food intake (i.e. appetite suppression). Based on preliminary clinical data, incretin mimetics and DPP-IV inhibitors show potential for treating type 2 diabetes. PMID- 15896684 TI - Bridging cheminformatic metabolite prediction and tandem mass spectrometry. AB - Despite recent technological advances, the analysis of biological samples for metabolite identification purposes often requires prior knowledge of the metabolite masses to successfully acquire high quality mass spectral data in the presence of intense background and interfering matrix signals. This, in turn, necessitates prior knowledge of the metabolite structure, which in most cases can be predicted on the basis of the potential routes of metabolism of those functional groups present in the molecule. The following discussion highlights the significance of knowledge of the metabolite mass in facilitating the detection and structural elucidation of drug metabolites. PMID- 15896685 TI - The continuing evolution of shotgun proteomics. AB - Shotgun proteomics has emerged as a powerful approach for the analysis of complex protein mixtures, including biofluids, tissues, cells, organelles or protein complexes. Having evolved from the integration of chromatography and mass spectrometry, innovations in sample preparation, multidimensional chromatography, mass spectrometry and proteomic informatics continually facilitate, enable and challenge shotgun proteomics. As a result, shotgun proteomics continues to evolve and enable new areas of biological research, and is beginning to impact human disease diagnosis and therapeutic intervention. PMID- 15896686 TI - Bioinformatic methods for integrating whole-genome expression results into cellular networks. AB - Extracting a comprehensive overview from the huge amount of information arising from whole-genome analyses is a significant challenge. This review critically surveys the state of the art methods that are used to connect information from functional genomic studies to biological function. Cluster analysis methods for inferring the correlation between genes are discussed, as are the methods for integrating gene expression information with existing information on biological pathways and the methods that combine cluster analysis with biological information to reconstruct novel biological networks. PMID- 15896690 TI - Human-centered computing in health information systems part 2: evaluation. PMID- 15896691 TI - Primary care physician time utilization before and after implementation of an electronic health record: a time-motion study. AB - Despite benefits associated with the use of electronic health records (EHRs), one major barrier to adoption is the concern that EHRs may take longer for physicians to use than paper-based systems. To address this issue, we performed a time motion study in five primary care clinics. Twenty physicians were observed and specific activities were timed during a clinic session before and after EHR implementation. Surveys evaluated physicians' perceptions regarding the EHR. Post implementation, the adjusted mean overall time spent per patient during clinic sessions decreased by 0.5 min (p=0.86; 95% confidence interval [-5.05, 6.04]) from a pre-intervention adjusted average of 27.55 min (SE=2.1) to a post intervention adjusted average of 27.05 min (SE=1.6). A majority of survey respondents believed EHR use results in quality improvement, yet only 29% reported that EHR documentation takes the same amount of time or less compared to the paper-based system. While the EHR did not require more time for physicians during a clinic session, further studies should assess the EHR's potential impact on non-clinic time. PMID- 15896692 TI - Identifying barriers to the effective use of clinical reminders: bootstrapping multiple methods. AB - Advances in electronic medical record capabilities enable clinical reminders to inform providers when recommended actions are "due" for a patient. Despite evidence that they improve adherence to guidelines, the Veteran's Health Administration (VHA) has experienced challenges in having providers consistently use clinical reminders as intended. In this paper, we describe how multiple methods were used to opportunistically triangulate, or "bootstrap," an understanding of barriers to the effective use of clinical reminders in the VHA. In an initial study using ethnographic observations and semi-structured interviews of HIV clinical reminders, we identified six barriers to effective use: workload, time to remove inapplicable reminders, false alarms, training, reduced eye contact, and the use of paper forms rather than software. In a second study, we collected open-ended and closed-ended data regarding barriers and facilitators to the use of clinical reminders in general in the VHA through a survey of 261 participants at a national informatics meeting, where 104 of 142 VHA health care facilities were represented. The findings from the second study extended our understanding of the previously identified barriers. In addition, four new barriers were identified: ease of use issues, accessibility of workstations, resident physicians and trainees, and administration benefiting more than providers from clinical reminder use. We discuss potential implications regarding the similarities and differences in study findings for factors to consider in planning interventions to improve clinical reminder use. PMID- 15896693 TI - Decisions about critical events in device-related scenarios as a function of expertise. AB - This paper presents the perspectives of personnel involved in decision-making about devices in critical care. We use the concept of "sharp and blunt ends" of practice to describe the performance of health care professionals. The "sharp end" is physically and temporally close to the system; the "blunt end" is removed from the system in time and space and yet affects the system through indirect influence on the sharp end. In this study, the sharp end is represented by the clinicians (nurses and doctors) and the blunt end by the administrators and biomedical engineers. These subjects represent the professionals involved in the decision-making process for purchasing biomedical equipment for the hospital. They were asked to "think aloud" while evaluating three error scenarios based on real events. The responses were recorded and transcribed for analysis. The results show differences in interpretation of critical events as a function of professional expertise. The clinicians (sharp-end practitioners) focused on clinical and human aspect of errors while the biomedical engineers focused on device-related errors. The administrators focused on documentation and training. These different interpretations mean that the problems are represented differently by these groups of subjects, and these representations result in variable decisions about devices. These results are discussed within a systems approach framework to help us assess the completeness of the problem representations of the subjects, their awareness of critical events, and how these events would collectively contribute to the occurrence of error. PMID- 15896694 TI - Human factors engineering: a tool for medical device evaluation in hospital procurement decision-making. AB - A human factors evaluation was conducted to inform hospital procurement decision making in selecting a general-purpose infusion pump to be used hospital-wide. Three infusion pumps from different vendors were involved in the evaluation, which consisted of two phases: a human factors heuristic assessment of the pumps according to several criteria, and user testing in five clinical areas. The clinical areas were: Oncology, Medical/Surgical, Pediatric, ICU, and Anaesthesiology. Fourteen nurses and three anaesthetists participated in the user testing. Reasonable agreement was observed between results of both phases of the evaluation, and overall results clearly favoured one of the infusion pumps over the others. It is recommended that a human factors evaluation should be performed to influence all hospital procurement decisions when purchasing medical devices, to ensure the best devices are selected for the end users and to ultimately enhance patient safety. PMID- 15896695 TI - Telephone-linked care for physical activity: a qualitative evaluation of the use patterns of an information technology program for patients. AB - Automated health behavior interventions that involve discretionary use by patients or consumers over extended periods of time are becoming more common and it is generally assumed that adherence to the recommended schedule is related to the impact of the system on users. Yet reasons for use or non-use of such systems have not been carefully explored. An understanding of factors that influence people to use, not use, or underutilize these automated behavioral change and self-care management systems can help in designing systems that are more effective and acceptable to users. Using qualitative research methods, this study explored the experiences of 45 users of a multiple-contact health promotion application with the goal of understanding the major factors that affect patterns of use (frequency of and duration of contact). The in-depth exploration of users' perceptions and views made possible by the qualitative research methods revealed a number of important themes. Reported reasons for underutilization or non-use were found to be both user-related and system-related. User-related reasons encompassed personal and individual events that prevented or impeded system utilization. System-related reasons included those that related to the medium itself as well as the content of the application. The qualitative methods employed in this study created a forum through which users' feedback could be fully explored and then synthesized to assist in the improvement of this and other automated health behavior interventions. PMID- 15896696 TI - Technology, work, and information flows: lessons from the implementation of a wireless alert pager system. AB - The combination of collaborative work practices and information technology affect the flow of information in clinical settings. The introduction of a new technology into these settings can change not only established work practices but also the information flows. In this paper, we examine the introduction of a wireless alerts pager in a surgical intensive care unit (SICU). Through a qualitative study, we analyze the effects that this new information tool had on both the work practices in the SICU and the information flow in the unit. We describe four challenges that SICU staff members faced with respect to the alerts pagers. We found that the pager provided new routes of information to SICU staff but in doing so disrupted existing work practices and information flows. PMID- 15896697 TI - Evaluation of a pulmonary graphical display in the medical intensive care unit: an observational study. AB - We developed a pulmonary graphic display that depicts pulmonary physiological variables for intubated, mechanically ventilated patients in a graphical format. The pulmonary graphical display presents multiple respiratory variables and changes are depicted by alterations in shape and color. Learning how this new technology will be integrated and accepted by users is an important step before it is introduced into the clinical arena. This study observed use and acceptance of the pulmonary graphical display by health care providers in an intensive care unit. Investigators noted that physicians, respiratory therapists, and nurses observed the pulmonary graphical display on average six, three, and one times, respectively, per patient room entry. Based on questionnaires, the pulmonary graphical display was perceived as useful, a desirable addition to current ICU monitors, and an accurate representation of respiratory variables. PMID- 15896698 TI - Evaluating user interactions with clinical information systems: a model based on human-computer interaction models. AB - OBJECTIVES: This article proposes a model for dimensions involved in user evaluation of clinical information systems (CIS). The model links the dimensions in traditional CIS evaluation and the dimensions from the human-computer interaction (HCI) perspective. PROPOSED METHOD: In this article, variables are defined as the properties measured in an evaluation, and dimensions are defined as the factors contributing to the values of the measured variables. The proposed model is based on a two-step methodology with: (1) a general review of information systems (IS) evaluations to highlight studied variables, existing models and frameworks, and (2) a review of HCI literature to provide the theoretical basis to key dimensions of user evaluation. RESULTS: The review of literature led to the identification of eight key variables, among which satisfaction, acceptance, and success were found to be the most referenced. DISCUSSION: Among those variables, IS acceptance is a relevant candidate to reflect user evaluation of CIS. While their goals are similar, the fields of traditional CIS evaluation, and HCI are not closely connected. Combining those two fields allows for the development of an integrated model which provides a model for summative and comprehensive user evaluation of CIS. All dimensions identified in existing studies can be linked to this model and such an integrated model could provide a new perspective to compare investigations of different CIS systems. PMID- 15896699 TI - Verbal coding and the storage of form-position associations in visual-spatial short-term memory. AB - Short-term memory for form-position associations was assessed using an object relocation task. Participants attempted to remember the positions of either three or five Japanese Kanji characters, presented on a computer monitor. Following a short blank interval, participants were presented with 2 alternative Kanji, only 1 of which was present in the initial stimulus, and the set of locations occupied in the initial stimulus. They attempted to select the correct item and relocate it back to its original position. The proportion of correct item selections showed effects of both articulatory suppression and memory load. In contrast, the conditional probability of location given a correct item selection showed an effect of load but no effect of suppression. These results are consistent with the proposal that access to visual memory is aided by verbal recoding, but that there is no verbal contribution to memory for the association between form and position. PMID- 15896700 TI - Antiparasitic activity of a triphenyl tin complex against Leishmania donovani. AB - Visceral leishmaniasis is a life-threatening human disease commonly known as kala azar. Leishmania donovani is the causative agent of this parasitic disease transmitted by the sand fly vector to infect hosts. Triphenyl tin salicylanilide thiosemicarbazone [Ph(3)Sn(OSal.TSCZH)] (TTST) which is an organometallic complex of triphenyl tin has been evaluated to explore possibility to develop a potent chemotherapeutic agent against visceral leishmaniasis. Effect of triphenyl tin complex on growth inhibition and host--parasite interaction were examined both in vitro and in vivo. Release of toxic superoxide radical was measured spectrophotometrically through the formation of blue formazan derived from reduced nitrobluetetrazolium. To understand mode of action of Ph(3)Sn(OSal.TSCZH), superoxide dismutase activity was determined spectrophotometrically by measuring ability of this enzyme to inhibit pyrogallol autoxidation and also by activity staining of the non-denaturing polyacrylamide gels after separating superoxide dismutase. Antileishmanial activity of triphenyl tin complex were found to be effective both in vitro and in vivo at lower concentrations compared to the existing toxic drugs available. IC(50) of Ph(3)Sn(OSal.TSCZH) was calculated as 0.05+/-0.01mg/L. Intracellular survival of the parasite in host macrophages was inhibited by TTST in a dose dependent manner. Parasite burden in spleen was reduced to 87% under TTST treatment (10mg/kg body weight) and under sodium antimony gluconate (20mg/kg body weight) reduced nearly to 65%. Its action as a chemotherapeutic agent is found to be mediated through inhibition of superoxide dismutase and simultaneous release of toxic superoxide radical. We propose that Ph(3) Sn(OSal.TSCZH) may be considered as a prospective candidate to establish a better line of therapeutic process against experimental visceral leishmaniasis. PMID- 15896701 TI - Protein kinase C-independent pathway for NADPH oxidase activation in guinea pig peritoneal polymorphonuclear leukocytes by cytochalasin D. AB - Cytochalasin D (CD) induced production of the superoxide radical (O(2)(-)) in guinea pig polymorphonuclear leukocytes (PMNs). The protein kinase C (PKC) inhibitor GF109203X (GFX) was rarely without effect on CD-induced O(2)(-) production. CD as well as PMA induced the translocation of p47(phox) to the membrane fraction, and this translocation was slightly decreased by GFX. Moreover, the inhibitory effect of a PKCzeta antagonist with sequences based on the endogenous PKCzeta pseudosubstrate region was weaker than the inhibitory effect on N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced O(2)(-) production. On the other hand, the production of O(2)(-) induced by CD was more strongly suppressed by the PLD inhibitor ethanol and phosphatidylinositol 3 kinase (PI3-K) inhibitor wortmannin than that induced by fMLP, and the activation of phospholipase D (PLD) by CD was restrained by wortmannin. These findings suggest that NADPH oxidase is activated by CD through a PKC-independent signaling pathway in PMNs, and this pathway involves the activation of PLD through PI3-K. PMID- 15896702 TI - Possible involvement of phosphatidylinositol 3-kinase/Akt signal pathway in vasopressin-induced HSP27 phosphorylation in aortic smooth muscle A10 cells. AB - We previously reported that p38 mitogen-activated protein (MAP) kinase takes a part in arginine vasopressin (AVP)-induced heat shock protein 27 (HSP27) phosphorylation in aortic smooth muscle A10 cells. In the present study, we investigated whether phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) is involved in the phosphorylation of HSP27 in these cells. AVP time-dependently induced the phosphorylation of PI3K and Akt. Akt inhibitor, 1l-6-hydroxymethyl chiro-inositol 2-(R)-2-O-methyl-3-O-octadecylcarbonate, partially suppressed the phosphorylation of HSP27. The AVP-induced HSP27 phosphorylation was attenuated by LY294002, a PI3K inhibitor. The combination of Akt inhibitor and SB203580, a p38 MAP kinase inhibitor, completely suppressed the AVP-induced phosphorylation of HSP27. Furthermore, LY294002 or Akt inhibitor did not affect the AVP-induced phosphorylation of p38 MAP kinase and SB203580 did not affect the phosphorylation of PI3K or Akt. These results suggest that PI3K/Akt plays a part in the AVP induced phosphorylation of HSP27, maybe independently of p38 MAP kinase, in aortic smooth muscle A10 cells. PMID- 15896703 TI - The kinase activity of human brain 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase is regulated via inhibition by phosphoenolpyruvate. AB - The two enzymatic activities of the highly conserved catalytic core of 6PF2K/Fru 2,6-P(2)ase are thought to be reciprocally regulated by the amino- and carboxy terminal regions unique to each isoform. In this study, we describe the recombinant expression, purification, and kinetic characterization of two human brain 6PF2K/Fru-2,6-P(2)ase splice variants, HBP1 and HBP2. Interestingly, both lack an arginine which is highly conserved among other tissue isoforms, and which is understood to be critical to the fructose-2,6-bisphosphatase mechanism. As a result, the phosphatase activity of both HBP isoforms is negligible, but we found that it could be recovered by restoration of the arginine by site directed mutagenesis. We also found that AMP activated protein kinase and protein kinases A, B, and C catalyzed the phosphorylation of Ser-460 of HBP1, and that in addition both isoforms are phosphorylated at a second, as yet undetermined site by protein kinase C. However, none of the phosphorylations had any effect on the intrinsic kinetic characteristics of either enzymatic activity, and neither did point mutation (mimicking phosphorylation), deletion, and alternative-splice modification of the HBP1 carboxy-terminal region. Instead, these phosphorylations and mutations decreased the sensitivity of the 6PF2K to a potent allosteric inhibitor, phosphoenolpyruvate, which appears to be the major regulatory mechanism. PMID- 15896704 TI - Self-association of glutamic acid-rich fusion peptide analogs of influenza hemagglutinin in the membrane-mimic environments: effects of positional difference of glutamic acids on side chain ionization constant and intra- and inter-peptide interactions deduced from NMR and gel electrophoresis measurements. AB - Two glutamic acid-rich fusion peptide analogs of influenza hemagglutinin were synthesized to study the organization of the charged peptides in the membranous media. Fluorescence and gel electrophoresis experiments suggested a loose association between the monomers in the vesicles. A model was built which showed that a positional difference of 3, 7 and 4, 8 results in the exposure of Glu3 and Glu7 side chains to the apolar lipidic core. Supportive results include: first, pK(a) values of two pH units higher than reference value in aqueous medium for Glu3 and Glu7 CgammaH, whereas the deviation of pK(a) from the reference value for Glu4 and Glu8 CgammaH is substantially smaller; second, Hill coefficients of titration shift of these protons indicate anti-cooperativity for Glu3 and Glu7 side chain protons but less so for Glu4 and Glu8, implying a strong electrostatic interaction between Glu3 and Glu7 possibly resulting from their localization in an apolar environment; third, positive and larger titration shift for NH of Glu3 is observed compared to that of Glu4, suggesting stronger hydrogen bond between the NH and the carboxylic group of Glu3 than that of Glu4, consistent with higher degree of exposure to hydrophobic medium for the side chain of Glu3. PMID- 15896705 TI - 2.0 A crystal structure of human ARL5-GDP3'P, a novel member of the small GTP binding proteins. AB - ARL5 is a member of ARLs, which is widespread in high eukaryotes and homologous between species. But no structure or biological function of this member is reported. We expressed, purified, and resolved the structure of human ARL5 with bound GDP3'P at 2.0 A resolution. A comparison with the known structures of ARFs shows that besides the typical features of ARFs, human ARL5 has specific features of its own. Bacterially expressed human ARL5 contains bound GDP3'P which is seldom seen in other structures. The hydrophobic tail of the introduced detergent Triton X-305 binds at the possible myristoylation site of Gly2, simulating the myristoylated state of N-terminal amphipathic helix in vivo. The structural features of the nucleotide binding motifs and the switch regions prove that ARL5 will undergo the typical GDP/GTP structural cycle as other members of ARLs, which is the basis of their biological functions. PMID- 15896706 TI - Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo. AB - In this study, we isolated CD31(-), CD34(-), CD106(-) (VCAM-1(-)), and fetal liver kinase(+) (Flk1(+)) cells from adipose tissue. These cells can be induced to differentiate into cells of osteogenic and adipogenic lineages in vitro and were termed adipose derived adult stem cells (ADAS cells). We also showed that they have characteristics of endothelial progenitor cells. In vitro, ADAS cells expressed endothelial markers when cultured with VEGF. In vivo, ADAS cells can differentiate in response to local cues into endothelial cells that contributed to neoangiogenesis in hindlimb ischemia models. PI3 kinase inhibitor LY294002 blocked the differentiation of ADAS cells into endothelial cells in vitro. Because ADAS cells can be expanded in culture without obvious senescence for more than 20 population doublings, they may be a potential source of endothelial cells for cellular pro-angiogenic therapies. PMID- 15896707 TI - Fucoxanthin from edible seaweed, Undaria pinnatifida, shows antiobesity effect through UCP1 expression in white adipose tissues. AB - Mitochondrial uncoupling protein 1 (UCP1) is usually expressed only in brown adipose tissue (BAT) and a key molecule for metabolic thermogenesis to avoid an excess of fat accumulation. However, there is little BAT in adult humans. Therefore, UCP1 expression in tissues other than BAT is expected to reduce abdominal fat. Here, we show reduction of abdominal white adipose tissue (WAT) weights in rats and mice by feeding lipids from edible seaweed, Undaria pinnatifida. Clear signals of UCP1 protein and mRNA were detected in WAT of mice fed the Undaria lipids, although there is little expression of UCP1 in WAT of mice fed control diet. The Undaria lipids mainly consisted of glycolipids and seaweed carotenoid, fucoxanthin. In the fucoxanthin-fed mice, WAT weight significantly decreased and UCP1 was clearly expressed in the WAT, while there was no difference in WAT weight and little expression of UCP1 in the glycolipids fed mice. This result indicates that fucoxanthin upregulates the expression of UCP1 in WAT, which may contribute to reducing WAT weight. PMID- 15896708 TI - Anc1 interacts with the catalytic subunits of the general transcription factors TFIID and TFIIF, the chromatin remodeling complexes RSC and INO80, and the histone acetyltransferase complex NuA3. AB - The Anc1 protein co-purifies with general transcription factors, chromatin remodeling complexes, and histone modification enzymes and is required for efficient transcription in yeast. We show here that Anc1 interacts with Tsm1, Tfg1, Sth1, Ino80, and Sas3 that are, respectively, the catalytic subunits of the general transcription factors TFIID and TFIIF, of the chromatin remodeling complexes RSC and INO80, and of the histone H3-acetyltransferase complex NuA3. We show that Anc1 is required for growth on galactose as the sole carbon source, and that it is recruited to the UAS of the GAL1 gene after induction. PMID- 15896709 TI - A gene expression fingerprint of mouse stomach ECL cells. AB - Many of the endocrine cells in the stomach are poorly characterized with respect to physiological significance. In some cases, the anticipated hormone has not yet been identified. Global gene expression analysis of mouse stomach was performed in an attempt to identify the ECL-cell peptide/protein. Specific functional activation (omeprazole-induced hypergastrinaemia) was used as a tool to generate a gene expression fingerprint of the ECL cells. The proposed fingerprint includes 14 genes, among them six are known to be expressed by ECL cells (=positive controls), and some novel ones, which are likely to be ECL-cell-related. The known ECL-cell-related genes are those encoding histidine decarboxylase, chromogranin A and B, vesicular monoamine transporter 2, synaptophysin, and the cholecystokinin-B receptor. In addition, the fingerprint included five genes, which might be involved in the process of secretion and three ESTs with unknown function. Interestingly, parathyroid hormone-like hormone (Pthlh) was identified as a candidate ECL-cell peptide hormone. PMID- 15896710 TI - The carboxy-terminal end of the peptide deformylase from Mycobacterium tuberculosis is indispensable for its enzymatic activity. AB - The peptide deformylase in bacteria is involved in removal of N-formyl group from newly synthesized proteins. The gene encoding this enzyme from Mycobacterium tuberculosis was cloned and expressed in Escherichia coli. The enzyme activity of the recombinant protein (mPDF) was insensitive to modulation by common monovalent/divalent cations. Kinetic analysis, using N-formylmethionine-alanine as the substrate, yielded K(cat)/K(m) of approximately 1220 M(-1)s(-1). Actinonin, a naturally occurring antibiotic, and 1,10-ortho-phenanthroline strongly inhibited the enzyme activity. The mPDF was very stable at 30 degrees C with a half-life of approximately 4h and exhibited resistance to oxidizing agents, like H(2)O(2). Thus, the mPDF achieved distinction in its behavior among any reported iron-containing peptide deformylases. Furthermore, amino acid sequence analysis of mPDF revealed the presence of an unusually long carboxy terminal end (residues 182-197), which is atypical for any gram-positive bacteria. Our results, through deletion analysis, for the first time established the role of this region in mPDF enzyme activity. PMID- 15896711 TI - Combination of 5-fluorouracil and genistein induces apoptosis synergistically in chemo-resistant cancer cells through the modulation of AMPK and COX-2 signaling pathways. AB - 5-Fluorouracil (5-FU) is one of the widely used chemotherapeutic drugs targeting various cancers, but its chemo-resistance remains as a major obstacle in clinical settings. In the present study, HT-29 colon cancer cells were markedly sensitized to apoptosis by both 5-FU and genistein compared to the 5-FU treatment alone. There is an emerging evidence that genistein, soy-derived phytoestrogen, may have potential as a chemotherapeutic agent capable of inducing apoptosis or suppressing tumor promoting proteins such as cyclooxygenase-2 (COX-2). However, the precise mechanism of cellular cytotoxicity of genistein is not known. The present study focused on the correlation of AMPK and COX-2 in combined cytotoxicity of 5-FU and genistein, since AMPK is known as a primary cellular homeostasis regulator and a possible target molecule of cancer treatment, and COX 2 as cell proliferation and anti-apoptotic molecule. Our results demonstrated that the combination of 5-FU and genistein abolished the up-regulated state of COX-2 and prostaglandin secretion caused by 5-FU treatment in HT-29 colon cancer cells. These appear to be followed by the specific activation of AMPK and the up regulation of p53, p21, and Bax by genistein. Under same conditions, the induction of Glut-1 by 5-FU was diminished by the combination treatment with 5-FU and genistein. Furthermore, the reactive oxygen species (ROS) was found as an upstream signal for AMPK activation by genistein. These results suggested that the combination of 5-FU and genistein exert a novel chemotherapeutic effect in colon cancers, and AMPK may be a novel regulatory molecule of COX-2 expression, further implying its involvement in cytotoxicity caused by genistein. PMID- 15896712 TI - Gir2 is an intrinsically unstructured protein that is present in Saccharomyces cerevisiae as a group of heterogeneously electrophoretic migrating forms. AB - Gir2 is a highly acidic cytoplasmic protein of Saccharomyces cerevisiae of unknown function that shows an anomalous migration on SDS-PAGE. Based on its large Stokes radius and thermostability, we have previously suggested that Gir2 lacks extensive secondary structure. Here we report that Gir2 is extremely sensitive to proteolysis when compared to glutathione-S-transferase, a highly structured protein, further indicating its unfolded nature. Prediction based on the FoldIndex program also indicates that Gir2 is a disordered protein. Using truncated forms of Gir2 we show that the N-terminal half of this protein, with its high content of acidic amino acid residues, is responsible for the anomalous electrophoretic behavior of Gir2. Because all these features are hallmarks of intrinsically unstructured proteins (IUP), we propose that Gir2 is another representative of the IUP group of proteins. Additionally, we describe that the endogenous yeast Gir2 shows heterogeneous electrophoretic mobility, which is not due to proteolytic cleavage. PMID- 15896713 TI - Epigenetic regulation of the dendritic cell-marker gene ADAM19. AB - Human ADAM19 (MADDAM) is a molecular marker for human dendritic cells and not expressed in macrophages. To investigate its cell-type-specific expression, we defined the transcriptional start site and the proximal promoter. Sequence analysis of the promoter revealed putative binding sites for several transcription factors including Sp1, Sp3, NF-kappaB, and VDR. A minimal promoter construct of 150 bp showed little difference in reporter activity between macrophages and dendritic cells. Transfection of monocytic THP-1 with the 150-bp fragment also resulted in significant reporter activity, despite the lack of endogenous MADDAM expression. TSA, a known inhibitor of histone deacetylation, led to a dose-dependent induction of MADDAM mRNA in THP-1. ChIP assays demonstrated high levels of acetylated histone H3 in the proximal promoter region of the MADDAM gene in TSA-treated THP-1 cells and dendritic cells as compared to macrophages, indicating an important role of histone acetylation in the regulation of the MADDAM gene. PMID- 15896714 TI - Genomic segments cloning and analysis of Cotesia plutellae polydnavirus using plasmid capture system. AB - Cotesia plutellae polydnaviruses (CpBV) has a segmented genome consisting of multiple circular double stranded DNAs. Recently, we have developed an easy, simple, and convenient system based on Tn7 transposition in order to clone genomic segments of CpBV in Escherichia coli cell and designated plasmid capture system (PCS). The PCS donor-S transferred a pUC19 origin of replication and an ampicillin resistance marker into CpBV genomic DNA by in vitro transposition. Through PCS system, we were able to clone 53 genomic clones ranging from 0.1 to 25.5 kb and further they were classified into 29 segments by their sizes and restriction endonuclease patterns. Among them, a complete nucleotide sequence of CpBV-S28 segment was determined and 10 putative genes were predicted from this segment. Interestingly, 9 of 10 putative ORFs had high level of similarities with catalytic domain of protein tyrosine phosphatase. Also, ORF2807 showed similarity with EP1-like proteins of C. congregata polydnavirus. PMID- 15896715 TI - Random mtDNA deletions and functional consequence in aged human skeletal muscle. AB - Mitochondrial respiratory chain deteriorates with age, mostly in tissues with high energy requirements. Damage to mitochondrial DNA (mtDNA) by reactive oxygen species is thought to contribute primarily to this impairment. However, the overall extent of random mtDNA mutations has still not been evaluated. We carried out molecular and biochemical analyses in muscle biopsies from healthy young and aged subjects. Deleted mtDNA accumulation was followed by both quantitative PCR analysis to quantify total mtDNA, and Southern-blotting, to determine deleted to full length mtDNA ratio. Enzymatic activities of the mitochondrial respiratory chain were measured in all subjects. Randomly deleted mtDNA appeared mainly in the oldest subjects (beyond 80 years old), affecting up to 70% of mtDNA molecules. The activities of complexes III and IV of the respiratory chain, complexes with mtDNA encoded subunits, are lower in the aged subjects. Physical activity could be one major parameter modulating the mitochondrial respiratory chain activity in aged muscle. PMID- 15896716 TI - Baculovirus p35 increases pancreatic beta-cell resistance to apoptosis. AB - beta-cells die by apoptosis in type 1 diabetes as a result of autoimmune attack mediated by cytokines, and in type 2 diabetes by various perpetrators including human islet amyloid polypeptide (hIAPP). The cascade of apoptotic events induced by cytokines and hIAPP is mediated through caspases and reactive oxygen species. The baculovirus p35 protein is a potent anti-apoptotic agent shown to be effective in a variety of species and able to inhibit a number of apoptotic pathways. Here, we aimed at determining the protective potential of p35 in beta cells exposed to cytokines and hIAPP, as well as the effects of p35 on beta-cell function. The p35 gene was introduced into betaTC-tet cells, a differentiated murine beta-cell line capable of undergoing inducible growth-arrest. Both proliferating and growth-arrested cells expressing p35 manifested increased resistance to cytokines and hIAPP, compared with control cells, as judged by cell viability, DNA fragmentation, and caspase-3 activity assays. p35 was significantly more protective in growth-arrested, compared with proliferating, cells. No significant differences were observed in proliferation and insulin content between cells expressing p35 and control cells. In contrast, p35 manifested a perturbing effect on glucose-induced insulin secretion. These findings suggest that p35 could be incorporated as part of a multi-pronged approach of immunoprotective strategies to provide protection from recurring autoimmunity for transplanted beta-cells, as well as in preventive gene therapy in type 1 diabetes. p35 may also be protective from beta-cell damage caused by hIAPP in type 2 diabetes. PMID- 15896717 TI - GITR overexpression on CD4+CD25+ HTLV-1 transformed cells: detection by massively parallel signature sequencing. AB - HTLV-I is the etiologic agent of adult T-cell leukemia (ATL), a fatal T-cell malignancy that is associated with profound immunosuppression. In this study, comprehensive gene expression profiling was performed using massively parallel signature sequencing (MPSS) to investigate virus-host interactions in acutely HTLV-1 transformed cells. The analysis revealed the modulation of numerous genes across different functional classes, many of which have not been previously implicated in HTLV-1 transformation or ATL. Differences in the transcriptomes of transformed cell lines were observed that have provided clues on how different clonal populations of cells respond to virus transformation. Quantitation of HTLV 1 transcription was possible, thus making MPSS a useful tool to study emerging pathogens and unknown microbial causes of human diseases. Importantly, overexpression of GITR, an activation marker that has not been previously reported to be upregulated by HTLV-1-infection or in transformed/leukemic cells and that is associated with the suppressor phenotype of CD4+CD25+ regulatory T cells (Tregs), was also observed. The deep and quantitative gene expression profile generated by MPSS should provide additional leads for discovery research that can be applied to better understand the pathobiology of HTLV-1 transformation and ATL as well as to developing new therapies. PMID- 15896718 TI - Model of Alzheimer's disease amyloid-beta peptide based on a RNA binding protein. AB - Although Alzheimer's Abeta peptide has been shown to form beta-sheet structure, a high-resolution molecular structure is still unavailable to date. A search for a sequence neighbor using Abeta(10-42) as the query in the Protein Data-Bank (PDB) revealed that an RNA binding protein, AF-Sm1 from Archaeoglobus fulgidus (PDB entry: 1i4k chain Z), shared 36% identical residues. Using AF-Sm1 as a template, we built a molecular model of Abeta(10-42) by applying comparative modeling methods. The model of Abeta(10-42) contains an antiparallel beta-sheet formed by residues 16-23 and 32-41. Hydrophobic surface constituted by residues 17-20 (LVFF) separates distinctly charged regions. Residues that interact with RNA in the AF-Sm1 crystal structure were found to be conserved in Abeta. Using a native gel we demonstrate for the first time that RNA can interact with Abeta and selectively retard the formation of fibrils or higher-order oligomers. We hypothesize that in a similar fashion to AF-Sm1, RNA interacts with Abeta in the beta-hairpin (beta-turn-beta) structure and prevents fibril formation. PMID- 15896719 TI - Systematic high-yield production of human secreted proteins in Escherichia coli. AB - Human secreted proteins play a very important role in signal transduction. In order to study all potential secreted proteins identified from the human genome sequence, systematic production of large amounts of biologically active secreted proteins is a prerequisite. We selected 25 novel genes as a trial case for establishing a reliable expression system to produce active human secreted proteins in Escherichia coli. Expression of proteins with or without signal peptides was examined and compared in E. coli strains. The results indicated that deletion of signal peptides, to a certain extent, can improve the expression of these proteins and their solubilities. More importantly, under expression conditions such as induction temperature, N-terminus fusion peptides need to be optimized in order to express adequate amounts of soluble proteins. These recombinant proteins were characterized as well-folded proteins. This system enables us to rapidly obtain soluble and highly purified human secreted proteins for further functional studies. PMID- 15896720 TI - Presenilin acts as a positive regulator of basal level activity of ERK through the Raf-MEK1 signaling pathway. AB - Presenilins (PS) have been reported to be functionally involved in amyloid precursor protein processing, notch receptor signaling, and programmed cell death, or apoptosis. To understand the role of PS1 in the signaling events, we investigated in this study the role of PS1 in the basal level of mitogen activated protein kinase (MAPK) pathways using PS1(-/-) mouse embryonic fibroblast (MEF) cells from PS1-null mice. Interestingly, the basal level of ERK activity, but not JNK or p38 activity, is lower in PS1(-/-) MEF cells than in PS1(+/+) MEF cells. In PS1(-/-) MEF cells, the basal activities of Raf and MEK, the upstream signaling component of ERK, are also lower than in PS1(+/+) MEF cells. Furthermore, Elk-1 transcription activity also down-regulates in PS1(-/-) MEF cells. Collectively, our data suggest that PS can modulate the basal level of ERK activity through the Raf-MEK-dependent pathway. PMID- 15896721 TI - Clinical evaluation and mitochondrial DNA sequence analysis in three Chinese families with Leber's hereditary optic neuropathy. AB - We report here the clinical, genetic, and molecular characterization of three Chinese families (WZ4, WZ5, and WZ6) with Leber's hereditary optic neuropathy (LHON). Clinical and genetic evaluations revealed the variable severity and age of-onset in visual impairment in these families. Penetrances of visual impairment in these Chinese families were 33.3%, 35.7%, and 35.5%, respectively, with an average 34.8%. Furthermore, the average age-at-onset in those Chinese families was 17, 20, and 18 years. In addition, the ratios between affected male and female matrilineal relatives in these Chinese families were 3:0, 1:1, and 1.2:1, respectively. Sequence analysis of the complete mitochondrial genomes in these pedigrees showed the distinct sets of mtDNA polymorphism, in addition to the identical G11778A mutation associated with LHON in many families. The fact that mtDNA of those pedigrees belonged to different haplogroups F1, D4, and M10 suggested that the G11778A mutation occurred sporadically and multiplied through evolution of the mtDNA in China. However, there was the absence of functionally significant mutations in tRNA and rRNAs or secondary LHON mutations in these Chinese families. The I187T mutation in the ND1, the S99A mutation in the A6, the V254I in CO3, and I58V in ND6 mutation, showing high evolutional conservation, may contribute to the phenotypic expression of the G11778A mutation in the WZ6 pedigree. By contrast, none of mtDNA variants are evolutionarily conserved and implicated to have significantly functional consequence in WZ4 and WZ5 pedigrees. Apparently, these variants do not have a potential modifying role in the development of visual impairment associated with G11778A mutation in those two families. Thus, nuclear modifier gene(s) or environmental factor(s) seem to account for the penetrance and expressivity of LHON in these three Chinese families carrying the G11778A mutation. PMID- 15896722 TI - The N-terminus of Drosophila ESC mediates its phosphorylation and dimerization. AB - The ESC protein, like other Polycomb Group proteins, is required for heritable silencing of the homeotic genes. ESC is phosphorylated in vivo, but the region of ESC that is phosphorylated and its consequences are not known. Here, we show that the amino-terminal region of ESC (residues 1-60) mediates its phosphorylation and dimerization. Phosphorylation of ESC1-60 in vitro by CK1 and CK2 strongly enhances its dimerization. Both phosphorylation and dimerization are conserved in the mammalian ESC homolog EED, suggesting that they play important roles in vivo. One role is suggested by the effect of phosphatase treatment on native ESC complexes, which does not affect the integrity of the 600 kDa ESC/E(Z) complex, but eliminates the 1 MDa ESC/E(Z) complex, which is distinguished from the former by the presence of the additional subunits PCL and RPD3. Thus, stability and perhaps assembly of larger ESC complexes may depend on ESC phosphorylation. PMID- 15896723 TI - Pharmacological inhibition of DA- and 5-HT activity blocks spontaneous and cocaine-activated behavior: reversal by chronic cocaine treatment. AB - Recently it was shown that the combined pretreatment with low autoreceptor preferring dose levels of apomorphine (0.05 mg/kg) and 8-OHDPAT (0.05 mg/kg), which decrease dopaminergic and serotonergic activity, induces a profound behavioral inhibition and also blocks the stimulant effects of cocaine. In two experiments, we report that the acute blockade of spontaneous and cocaine locomotor stimulant effects by pretreatment with 8-OHDPAT (0.05 mg/kg) plus apomorphine (0.05 mg/kg) is dose-dependently (0.0 2.5, 5.0, and 10.0 mg/kg cocaine) reversed with repeated cocaine treatments. Using a paired vs. unpaired Pavlovian conditioning protocol, we found that this reversal by cocaine (10 mg/kg) of the inhibition by the combined 8-OHDPAT plus apomorphine pretreatment occurred for the paired but not the unpaired cocaine treatment. The findings suggest that this reversal of behavioral inhibition is mediated by the transformation of the drug cues generated by 8-OHDPAT and apomorphine into cocaine-conditioned stimuli which can activate behavior. PMID- 15896724 TI - Activation of presynaptic GABAA receptors increases spontaneous glutamate release onto noradrenergic neurons of the rat locus coeruleus. AB - In order to further explore how GABA can modulate the excitability of noradrenergic neurons of the locus coeruleus (LC), we investigated the presence of GABA(A) receptors on glutamatergic nerve terminals and the functional consequences of their activation. We used mechanically dissociated immature rat LC neurons with adherent nerve terminals and patch-clamp recordings of spontaneous excitatory postsynaptic currents. Activation of presynaptic GABA(A) receptors by muscimol facilitated spontaneous glutamate release by activating tetrodotoxin-sensitive Na(+) channels and high-threshold Ca(2+) channels. Bumetanide (10 microM), a potent blocker of Na(+)-K(+)-Cl(-) cotransporter, diminished the muscimol-induced facilitatory action of glutamate release. Our results indicate that the Na(+)-K(+)-Cl(-) cotransporter accumulates Cl(-) inside the nerve terminals so that activation of presynaptic GABA(A) receptors causes depolarization. This GABA(A)-receptor-mediated modulation of spontaneous glutamatergic transmission is another mechanism by which GABA and its analogues can regulate the excitability and activity of noradrenergic neurons in the LC. PMID- 15896725 TI - Phenotypic profile of SWR/J and A/J mice compared to control strains: possible mechanisms underlying resistance to obesity on a high-fat diet. AB - To understand mechanisms underlying a resistance to obesity, two obesity resistant inbred mouse strains, SWR/J and A/J, were compared to 3 inbred "control" strains, C3H/HeJ, BALB/cByJ and C57L/J. These 5 strains, studied at 5 weeks of age when similar in body weight, were maintained for 3 weeks on a 3-diet feeding paradigm, with separate jars of carbohydrate, protein and fat, or for 1 week on a single high-fat or low-fat diet. The control strains each chose a balanced diet, with 50% carbohydrate and 15-25% fat, and they had a similar, normal range of scores for measures of body weight, adiposity, endocrine parameters and metabolic enzyme activity. Compared to these control strains, the obesity-resistant SWR/J and A/J strains consumed more total calories and selected a diet with significantly more fat (35-45%) and less carbohydrate (35%). Despite overeating, they weighed less and had significantly reduced adiposity. They also had lower levels of insulin and exhibited increased capacity of skeletal muscle to metabolize fat, as indicated by measures beta-hydroxyacyl-CoA dehydrogenase activity or its ratio to citrate synthase. Measurements of hypothalamic peptides via radioimmunoassay or real-time quantitative PCR revealed markedly enhanced galanin (GAL) in the paraventricular nucleus and reduced neuropeptide Y (NPY) expression in the arcuate nucleus of obesity-resistant mice. These patterns in SWR/J and A/J strains, seen on a low-fat as well as high-fat diet, may reflect mechanisms involving excess GAL and reduced NPY that contribute early, respectively, to the over-consumption of a high-fat diet and a resistance to the obesity-promoting effects of this diet. PMID- 15896726 TI - Expression of the vanilloid receptor TRPV1 in rat dorsal root ganglion neurons supports different roles of the receptor in visceral and cutaneous afferents. AB - A combination of tracing and multiple color immunofluorescence revealed that 69% of rat dorsal root ganglion (DRG) neurons innervating the urinary bladder expressed the vanilloid receptor TRPV1. In contrast, only 32% of DRG neurons innervating the skin of the L6 dermatome expressed TRPV1. However, a similar fraction of visceral (60-62%) and of cutaneous (59-60%) TRPV1-positive DRG neurons expressed the peptidergic markers substance P and calcitonin gene-related peptide, while the fraction of TRPV1-positive neurons that was labeled by the non peptidergic marker Isolectin B4 was 58% for cutaneous and only 24% for visceral afferents. These results underscore differences of expression of functional markers in visceral and cutaneous afferents and support different mechanisms of activation of TRPV1 in viscera and in skin. PMID- 15896727 TI - Myocyte apoptosis in heart failure. AB - Human heart failure is preceded by a process termed cardiac remodeling in which heart chambers progressively enlarge and contractile function deteriorates. Programmed cell death (apoptosis) of cardiac muscle cells has been identified as an essential process in the progression to heart failure. The execution of the apoptotic program entails complex interactions between and execution of multiple molecular subprograms. Unlike necrosis, apoptosis is an orderly regulated process and, by inference, a logical therapeutic target if intervention occurs at an early stage. To identify potential therapeutic targets, it is imperative to have a full understanding of the apoptotic pathways that are functional in the cardiac muscle. Accordingly, the present review summarizes the apoptotic pathways operative in cardiac muscle and discusses therapeutic options related to apoptosis for the future treatment of human heart failure. PMID- 15896728 TI - Falsely low LDL-cholesterol concentrations and artifactual undetectable HDL cholesterol measured by direct methods in a patient with monoclonal paraprotein. AB - INTRODUCTION: Multiple myeloma is a malignant immunoproliferative disorder with lipoprotein abnormalities. We report a case of falsely low concentrations of LDL cholesterol (LDL-C) and artifactural undetectable HDL-cholesterol (HDL-C) as measured with direct methods in a patient of multiple myeloma with IgGkappa monoclonal gammapathy and significant hyperlipidemia. CASE REPORT: The patient had HDL-C and LDL-C concentrations in the 0.63-0.71 mmol/l and 2.22-2.36 mmol/l ranges, respectively, as measured by a traditional semi-quantitative electrophoresis method. The observation of falsely low concentrations of LDL-C and artifactural undetectable HDL-C might result in the mismanagement of patients of multiple myeloma with monoclonal gammapathy, because the LDL-C and HDL-C concentrations are positive and negative risk factors of cardiovascular diseases. CONCLUSIONS: Care must be taken when using the homogenous method for direct measurement of LDL-cholesterol and HDL-cholesterol in patients of multiple myeloma with monoclonal paraprotein. PMID- 15896729 TI - The clinical application of proteomics. AB - BACKGROUND: Proteomics is defined as a scientific approach used to elucidate all protein species within a cell or tissue, and many researchers are taking advantage of proteomic technology to elucidate protein changes between healthy and diseased states. METHODS: The application of proteomic techniques and strategies to the field of medicine is slowly transforming the way biomarker discovery is conducted. However, the complexity of serum is the source of both its promise to clinical applications and its challenge to proteomic analysis. Like any new technology when it is first introduced, proteomics has been touted with much hope and promise. RESULTS AND CONCLUSIONS: We provide a review of the clinical application of proteomics with the emphasis on current practical issues and challenges facing proteomic research. PMID- 15896730 TI - Proteomic: new advances in the diagnosis of rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) is a chronic autoimmune disease affecting the joints. A number of novel treatment modalities have been introduced over the past years, and rheumatologists are now attempting to institute optimal treatment in recent-onset arthritis. To facilitate diagnosis during the early stages of disease, when often not all clinical symptoms are manifest, a good serological marker is needed. METHODS: Antibodies directed to citrullinated proteins provide this ability. The most sensitive assay for detecting these antibodies is the so called anti-cyclic citrullinated peptide, second generation (CCP II) enzyme linked immunosorbent assay (ELISA). RESULTS: The diagnostic and prognostic potential of anti-CCP antibodies and the availability of a fully automated assay method lead us to conclude that the test is satisfactory for routine use as a serological marker of RA. In addition, we consider the potential of multiplex autoantibody assays, including miniaturized, high-throughput microarray technology, to improve diagnosis and prognostication in early onset arthritis patients. PMID- 15896731 TI - Differential effects of 5-HT2C receptor ligands on place conditioning and locomotor activity in rats. AB - Effects of the 5-hydroxytryptamine (5-HT)(1A/1B/2C) receptor agonist N-[3 (trifluoromethyl)phenyl] piperazine (TFMPP, 0-3.0 mg/kg s.c.) and the 5-HT2C receptor agonist 8,9-dichloro-2,3,4,4a-tetrahydro-1H-pyrazino[1,2-a]quinoxalin 5(6H)-one (WAY 161503, 0-3.0 mg/kg s.c.) in place conditioning were measured in male Sprague-Dawley rats. Effects of TFMPP, alone and with the 5-HT(1A) receptor antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl] ethyl]-N-2-pyridinyl cyclohexanecarboxamine (WAY 100635), the 5-HT(1B) receptor antagonist N-[4 methoxy-3-(4-methyl-1-piperazinyl) phenyl]-2'-methyl-4'-(5-methyl-1,2,4-oxadiazol 3-yl)-1,1'-biphenyl-4-carboxamide (GR 127935) or the 5-HT2C receptor antagonist 6 chloro-5-methyl-1-[[2-(2-methylpyrid-3-yloxy)pyrid-5-yl]carbamoyl]indoline (SB 242084) and of WAY 161503 alone and with SB 242084 on locomotor activity were also assessed. Neither TFMPP nor WAY 161503 induced place conditioning. WAY 161503 (1.0 and 3.0 mg/kg s.c.) decreased locomotor activity; SB 242084 (1.0 mg/kg i.p.) blocked this effect. Reduced locomotor activity following TFMPP was blocked by SB 242084 but not WAY 100635 (0.1 mg/kg s.c.) or GR 127935 (3.0 mg/kg s.c.). Behaviourally relevant levels of 5-HT2C receptor stimulation may not exert reinforcing effects, although other studies indicate that such manipulations alter reinforcing effects of drugs of abuse. PMID- 15896732 TI - Nicotine physical dependence in the mouse: involvement of the alpha7 nicotinic receptor subtype. AB - Although chronic nicotine produces dependence in mice, the role of specific nicotinic receptors has not been examined in knockout animals. The present study utilized alpha7 nicotinic receptor knockout mice to explore the role of this receptor subunit in nicotine dependence. Mice were chronically exposed to nicotine (0 or 200 microg/ml) in their drinking water and assayed for somatic withdrawal signs, hyperalgesia (tail-flick and hot-plate) and spontaneous activity following nicotine cessation. Nicotine withdrawal produced increased somatic signs in both strains and hyperalgesia in wild-type, but not in knockout animals. These results indicate that the alpha7 nicotinic receptor subunit may mediate some aspects of nicotine dependence. PMID- 15896733 TI - Induction of human cholesterol 7alpha-hydroxylase in HepG2 cells by 2,4,6 trihydroxyacetophenone. AB - In animal the plasma cholesterol-lowering activity of 2,4,6 trihydroxyacetophenone (THA) is due to enhanced cholesterol 7alpha-hydroxylase (CYP7A1) activity. We have examined the effect of THA on CYP7A1 activity and mRNA level in HepG2 cells. THA stimulated CYP7A1 activity in a concentration- and time dependent manner. After exposure for 24 h, 1 muM THA induced CYP7A1 activity 160+/-8% and mRNA level 166+/-21% (mean+/-S.E.M.) of control. Moreover THA antagonized the inhibitory regulation of chenodeoxycholic acid on CYP7A1 mRNA expression. These results indicated that THA increases CYP7A1 activity in human HepG2 cells by stimulating mRNA transcription. PMID- 15896734 TI - [Nphe1,Arg14,Lys15]N/OFQ-NH2 is a competitive antagonist of NOP receptors in the periaqueductal gray. AB - Nociceptin/orphanin FQ (N/OFQ) and N/OFQ peptide (NOP) receptors are implicated in many physiological functions including pain regulation. This study quantitatively investigated the interaction of a novel NOP receptor antagonist, UFP-101 ([Nphe1,Arg14,Lys15]N/OFQ-NH2), with N/OFQ in the ventrolateral periaqueductal gray, a crucial midbrain area for pain regulation. N/OFQ concentration-dependently activated G-protein coupled inwardly rectifying K+ (GIRK) channels in ventrolateral neurons of periaqueductal gray slices. UFP-101 antagonized N/OFQ-induced GIRK channel activation in a concentration-dependent manner and produced a parallel shift of the concentration-response curve of N/OFQ. The pA2 value estimated from Schild plot is 6.92+/-0.06. At concentrations up to 1 microM, UFP-101 had no effect on membrane current per se and did not affect the GIRK current activated by [d-Ala2, N-Me-Phe4, Gly-ol5]-enkephalin, a mu-opioid receptor agonist. It is concluded that UFP-101 is a potent and competitive peptide antagonist of NOP receptors that mediate GIRK channel activation in ventrolateral periaqueductal gray neurons. PMID- 15896736 TI - How an enzyme answers multiple-choice questions. AB - Acetohydroxyacid synthase (AHAS) is the first common enzyme in the pathway for the biosynthesis of branched-chain amino acids. Interest in the enzyme has escalated over the past 20 years since it was discovered that AHAS is the target of the sulfonylurea and imidazolinone herbicides. However, several questions regarding the reaction mechanism have remained unanswered, particularly the way in which AHAS "chooses" its second substrate. A new method for the detection of reaction intermediates enables calculation of the microscopic rate constants required to explain this phenomenon. PMID- 15896737 TI - Deoxyribonucleoside kinases: two enzyme families catalyze the same reaction. AB - Mammals have four deoxyribonucleoside kinases, the cytoplasmic (TK1) and mitochondrial (TK2) thymidine kinases, and the deoxycytidine (dCK) and deoxyguanosine (dGK) kinases, which salvage the precursors for nucleic acids synthesis. In addition to the native deoxyribonucleoside substrates, the kinases can phosphorylate and thereby activate a variety of anti-cancer and antiviral prodrugs. Recently, the crystal structure of human TK1 has been solved and has revealed that enzymes with fundamentally different origins and folds catalyze similar, crucial cellular reactions. PMID- 15896738 TI - A putative nuclear function for mammalian Staufen. AB - In addition to its role in rRNA processing and ribosome assembly, the nucleolus plays a part in the assembly of non-ribosomal ribonucleoprotein particles (RNPs) that are destined for cytoplasmic RNA delivery. Recent evidence indicates that mammalian Staufen2, a brain-specific RNA-binding protein involved in RNA localization, can--at least transiently--enter the nucleolus. Therefore, the assembly of Staufen2 into transport-competent RNPs might occur in the nucleus before their export into the cytoplasm. This could provide new insights into the mechanisms of subcellular RNA localization. PMID- 15896739 TI - DNA aptamers as potential anti-HIV agents. AB - Guanine (G)-rich DNA sequences can adopt stable G-quadruplex structures by G tetrad hydrogen-bonding and hydrophobic stacking. Recently, it has been shown that a DNA sequence forms an aptamer (termed 93del) and adopts a novel dimeric quadruplex folding topology in K+ solution. This aptamer exhibits anti-HIV1 integrase activity in the nanomolar range in vitro. A docking-based model of the 93del-integrase complex positions the DNA aptamer within a channel of the tetrameric integrase. This mutual fitting blocks several catalytic amino acid residues that are essential for integrase function, and accounts for the anti HIV1 activity of the 93del aptamer. PMID- 15896740 TI - Mediator and the mechanism of transcriptional activation. AB - Mediator was discovered because of its activity in a yeast RNA polymerase II (pol II) transcription system - it is needed for the system to respond to a transcriptional activator. Mediator is the central link in the enhancer- >activator-->Mediator-->pol II-->promoter pathway. The transduction of regulatory signals through this pathway is crucial for transcription of almost all pol II promoters in all eukaryote organisms. PMID- 15896741 TI - The yeast Mediator complex and its regulation. AB - The Mediator complex acts as a bridge, conveying regulatory information from enhancers and other control elements to the basal RNA polymerase II transcription machinery. Mediator is required for the regulated transcription of nearly all RNA polymerase II-dependent genes in Saccharomyces cerevisiae, and post-translational modifications of specific Mediator subunits can affect global patterns of gene transcription. PMID- 15896742 TI - Interactions between subunits of Drosophila Mediator and activator proteins. AB - Mediator was first identified because of its activity in activator-stimulated transcription in vivo and in vitro. Later, biochemical fractionation led to the co-purification of the multi-subunit Mediator complex and RNA polymerase II (pol II). Results of these studies suggested a model whereby transcription-activator proteins, which bind to specific gene regulatory sequences, recruit both Mediator and pol II as a holoenzyme in a one-step mechanism. More recent studies of Drosophila Mediator and additional studies in yeast have demonstrated that different transcription activators can bind and recruit Mediator to promoters in vivo in a step that is independent of pol II recruitment. Moreover, the different activators in Drosophila bind and recruit Mediator via physical interactions with specific subsets of proteins. These features of Mediator function seem to be broadly conserved. PMID- 15896743 TI - The mammalian Mediator complex and its role in transcriptional regulation. AB - Mediator is an essential component of the RNA polymerase II general transcriptional machinery and plays a crucial part in the activation and repression of eukaryotic mRNA synthesis. The Saccharomyces cerevisiae Mediator was the first to be defined and is a high molecular mass complex composed of >20 distinct subunits that performs multiple activities in transcription. Recent studies have defined the subunit composition and associated activities of mammalian Mediator, and revealed a striking evolutionary conservation of Mediator structure and function from yeast to man. PMID- 15896744 TI - Dynamic regulation of pol II transcription by the mammalian Mediator complex. AB - Mammalian Mediator is a key coactivator that enables transcriptional activators to regulate transcription by RNA polymerase II (pol II). Like the yeast complex to which it is phylogenetically related, it contains up to 30 subunits. These subunits are organized as a tightly associated core sub-complex, which associates with several groups of subunits that might constitute distinct modules. Although the complex seems to be universally required at all genes, specific subunits are dedicated to regulation of distinct expression programs via interactions with relevant gene-specific transcriptional activators. These interactions, in conjunction with dynamic effects of the core complex on pol II and the general transcription factors, lead to activation of transcription at the target gene. In addition, the compositional complexity of the Mediator allows for assimilation of other diverse signals such as those emanating from repressors and other coactivators. PMID- 15896745 TI - Structure of eukaryotic Mediator complexes. AB - Mediator, a macromolecular complex comprising approximately 20 different protein components, is largely responsible for the tight control of transcription that underpins cell development, differentiation, and maintenance in eukaryotes from yeast to human. In the past five years, macromolecular electron microscopy has been used to characterize the structure of Mediator, and of the complexes it forms with other components of the transcription machinery. The results reveal how Mediator interacts with RNA polymerase II, and suggest that regulatory information could be conveyed through changes in Mediator conformation that would influence the transcription initiation process. PMID- 15896746 TI - "Stealth immune tolerance" in stem cell transplantation: potential for "universal donors" in myocardial regenerative therapy. PMID- 15896747 TI - Three-year results of a randomized, double-blind, controlled trial of mycophenolate mofetil versus azathioprine in cardiac transplant recipients. AB - BACKGROUND: This study reports the 36-month results of a randomized, double blind, active-controlled trial of mycophenolate mofetil (MMF) vs azathioprine (AZA) in heart transplant patients. METHODS: Patients were randomized at the time of transplant to receive MMF (1,500 mg twice a day, N = 327) or AZA (1.5 to 3 mg/kg in 4 daily doses, N = 323) in addition to cyclosporine and corticosteroids; 289 patients in each group received study drug. Data were analyzed in all randomized patients (enrolled) and in patients who received study medications (treated). Clinical and graft assessments continued for 36 months. RESULTS: For the co-primary end-point, 53 of 289 (18.3%) AZA-treated patients either died or received another transplant compared with 34 of 289 (11.8%) MMF-treated patients (p < 0.01). Time to re-transplantation or patient death was significantly shorter for AZA- than MMF-treated patients (p = 0.029). In patients undergoing intravascular ultrasound, the change in mean maximal intimal thickness was less for the MMF group than for the AZA group (0.06 +/- 0.03 mm vs 0.13 +/- 0.03 mm, respectively; p = 0.056). No significant differences between treatments were observed in quantitative coronary angiographic measurements of transplant coronary vasculopathy. Congestive heart failure, atrial arrhythmia and leukopenia were more common in the AZA group, whereas diarrhea, esophagitis, Herpes simplex, Herpes zoster and cytomegalovirus (CMV) tissue invasion were more common in MMF treated patients. CONCLUSION: MMF reduces mortality and graft loss up to 36 months after transplantation. PMID- 15896748 TI - Effect of long-term calcitonin administration on steroid-induced osteoporosis after cardiac transplantation. AB - BACKGROUND: Early, rapid bone loss and fractures after cardiac transplantation are well-documented complications of steroid administration; therefore, we undertook this study on the effects of long-term calcitonin on steroid-induced osteoporosis. METHODS: Twenty-three heart transplant recipients on maintenance immunosuppression with cyclosporine, mycophenolate mofetil and prednisone were retrospectively studied. All patients received long-term prophylactic treatment with elemental calcium and vitamin D. Twelve (52.2%) patients also received long term intranasal salmon calcitonin, whereas 11 (47.8%) received none. Bone mineral density and vertebral fractures were assessed at yearly intervals. Statistical comparisons between each group's bone loss during the first year and in the early (1 to 3 years), intermediate (4 to 6 years) and late (7+ years) post transplantation periods were done. RESULTS: Lumbar spine bone loss was significant during the early follow-up period in the group not receiving calcitonin (0.744 +/- 0.114 g/cm(2) vs 0.978 +/- 0.094 g/cm(2) [p = 0.002]). The calcitonin group showed bone mineral density (BMD) levels within normal average values throughout the study period. BMD increased in the no-calcitonin group during the intermediate (4 to 6 years) and late (7+ years) follow-up periods, with values approaching normal average and no significant difference between the 2 groups (0.988 +/- 0.184 g/cm(2) vs 0.982 +/- 0.088 g/cm(2) [p = 0.944] and 0.89 +/- 0.09 g/cm(2) vs 1.048 +/- 0.239 g/cm(2) [p = 0.474], respectively). CONCLUSIONS: Prophylactic treatment with intranasal salmon calcitonin prevents rapid bone loss associated with high-dose steroids early after cardiac transplantation. Long-term administration does not seem warranted in re establishing BMD. PMID- 15896749 TI - A descriptive study of small airway reticular basement membrane thickening in clinically stable lung transplant recipients. AB - BACKGROUND: Chronic rejection functionally manifested by fixed airflow limitation, bronchiolitis obliterans syndrome (BOS), is a major problem for all lung allograft programs. The inclusion of a pre-BOS category (BOS(0 approximately p)) in the newly revised guidelines, recognizes the potential importance of early changes. We tested the hypothesis that small airway reticular basement membrane thickening exists even in clinically stable lung transplant recipients with some evidence of inflammation but who are BOS-free. METHODS: A bronchoscopic study was performed on 30 clinically stable lung allograft recipients at >/=3 months post allograft, who were BOS-free but with some evidence of airway inflammation indicated by a pathologic diagnosis of lymphocytic bronchiolitis or raised exhaled nitric oxide (NO). After baseline physiologic assessment, small airway reticular basement membrane (Rbm) thickening was quantified in transbronchial biopsy (TBB) using image analysis, with inflammation assessed by bronchoalveolar lavage (BAL) differential cell counts. RESULTS: Twenty-one patients had technically satisfactory measurements of Rbm thickness. We detected small airway Rbm thickening when compared with published data for control lung diseases. There was no correlation between Rbm thickening and lung function (forced expiratory volume in 1 second [FEV(1)] best post-operatively and Rbm r = -0.10, not significant). CONCLUSIONS: Our data suggest that airway remodeling can occur early in lung allografts and before development of airflow limitation and BOS. Longitudinal pathophysiologic studies are needed to elucidate potential relationships between airway inflammation, Rbm thickening and allograft failure. Airway biopsies would be of value in such studies. PMID- 15896750 TI - Effects of inhaled FK 506 on the suppression of acute rejection after lung transplantation: use of a rat orthotopic lung transplantation model. AB - BACKGROUND: FK 506 inhalant was recently developed for localized administration. We investigated its effects on acute lung allograft rejection and compared its efficacy with that of intramuscular administration of FK 506. METHODS: Rats (n = 123) with orthotopic left lung transplantation were divided into 9 groups. Six groups inhaled FK 506 (5 puffs, 10 puffs or 20 puffs per day), or were given intramuscular administration of FK 506 (0.05, 0.1 or 1.0 mg/kg/day). The other groups included rats receiving an isograft, rats with an untreated allograft, and a placebo group. All groups (n = 6 each) were monitored for 14 days post operatively as an end-point and graft survival time was determined. The remaining animals were killed 4 days after transplantation. The histologic grade of rejection was determined for all groups (n = 6 each). With both (n = 3 each) inhalation therapy and intramuscular administration of FK 506, which showed similar degrees of effectiveness, both blood FK 506 concentration and cytokine expression in the graft and spleen were evaluated. RESULTS: FK 506 inhalation therapy extended allograft survival time and reduced histologic rejection on Day 4 in all groups. Graft survival time and histologic rejection scores at a dose of 10 puffs/day were comparable to those with 0.1 mg/kg/day of intramuscular FK 506. Trough concentrations of FK 506 in blood were detectable with 0.1 mg/kg/day of intramuscular FK 506, but not with 10 puffs/day. The messenger RNA expression levels of interferon-gamma in the lung allograft was suppressed significantly at a dose of 10 puffs/day. CONCLUSIONS: FK 506 inhalant enhances acute lung allograft survival with lower blood concentrations than when using comparable intramuscular administration. PMID- 15896751 TI - Diagnostic and prognostic value of dobutamine thallium-201 single-photon emission computed tomography after heart transplantation. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV) is a major cause of mortality in heart transplant recipients. We investigated the diagnostic and prognostic value of dobutamine thallium-201 ((201)Tl) single-photon emission computed tomography (SPECT) in heart transplant recipients. METHODS: We studied 47 patients (age 51.6 +/- 11.7 years, 37 men), at a mean of 34.0 +/- 21.4 months after heart transplant, who received dobutamine (201)Tl SPECT, echocardiography and coronary angiography within 1 month of each other. SPECT was considered abnormal in the presence of reversible or fixed defects in >/=2 segments. Significant CAV was defined as >/=50% luminal stenosis. RESULTS: Coronary angiograms were normal in 37 patients. Non-significant CAV was detected in 1 patient and significant CAV in 9 patients. The sensitivity, specificity, positive predictive value and negative predictive value of SPECT for the detection of significant angiographic CAV were 89%, 71%, 42% and 96%, respectively. Large reversible perfusion defects (>/=6 segments) always indicated significant CAV. In patients with normal left ventricular function, a lung/heart ratio (LHR) of >/=0.37 during stress was also an independent predictor of significant CAV (odds ratio 15.5, p = 0.04). A higher stress LHR was associated with greater vessel involvement (r = 0.516, p = 0.0002). Patients with impaired left ventricular function also had higher stress and resting LHR. Over 40.3 +/- 21.9 months after the first SPECT, 1 patient developed significant angiographic CAV and another 4 had cardiac death. Large reversible perfusion defect was a significant predictor of cardiac death (p = 0.002). CONCLUSIONS: Dobutamine (201)Tl SPECT is a useful method for detecting patients with significant CAV and assessing prognosis. It is reasonable and safe to design individualized surveillance intensity of coronary angiography for post transplant patients on the basis of non-invasive monitoring of dobutamine (201)Tl testing. PMID- 15896752 TI - Angiogenesis occurs within the intimal proliferation that characterizes transplant coronary artery vasculopathy. AB - BACKGROUND: Vascular remodeling is central to the development of transplant coronary artery vasculopathy (CAV). For remodeling to occur, a sustained blood and nutrient supply is essential. Here we report on the presence of angiogenesis within the neointima of coronary arteries from cardiac transplant recipients. METHODS: Coronary arteries from 57 cardiac transplant recipients with CAV were analyzed. Immunocytochemistry with antibodies raised against endothelial cells (CD31, CD34, and vWF), vascular smooth muscle cells (SmA), and activated endothelial cells (MHC 2, P-SEL, E-SEL, and VCAM-1) was performed. RESULTS: A total of 89% of patients had significant angiogenesis. These vessels appeared as endothelial lined channels and were present in a concentric circumferential pattern within the mid portion of the neointima. These new vessels were present at an interface between an area of intimal hyperplasia and below an area of fibrous regeneration. These 2 distinct zones were present in 64% of the cases, and were clearly demonstrated with an elastic van Gieson (EVG) stain and are distinctly different from that seen in native atherosclerosis. Endothelial activation markers were strongly expressed by the endothelial cells lining new vessels, suggesting that they are functional and may aid in the recruitment of inflammatory cells. CONCLUSIONS: These data suggest that angiogenesis is present within the intima of CAV lesions and may contribute to the continued obliteration of the vessel lumen. The vessels appear to originate in the intima and may represent the location of the donor endothelium before transplantation. Inhibition of endothelial damage may provide therapeutic options to prevent the progression of CAV. PMID- 15896753 TI - Interleukin-6 polymorphism: a genetic risk factor for cardiac transplant related coronary vasculopathy? AB - AIMS: Interleukin- (IL-) 6 is a pleiotropic cytokine with effects on the acute phase response, inflammation, and vascular function. A G to C polymorphism has been described at position -174 in the promoter region of the IL-6 gene. We investigated the influence of this polymorphism on the development of cardiac transplant related coronary vasculopathy (CV). METHODS: Sequence specific polymerase chain reaction identified the -174*G/C allele for 116 cardiac transplant recipients. Coronary disease was identified by routine surveillance post-transplant coronary angiography. RESULTS: Prevalence of the -174*G/C polymorphism was different between the transplant and control cohorts; *CC 27.6%, *CG 45.7%, and *GG 26.7% vs. 13.2%, 44.1% and 42.7% respectively (p = 0.004). Median time to the first diagnosis of CV was different between the 3 alleles; *CC 2.8 years (2.0-4.0); *CG 3.9 years (2.1-4.5); *GG 5.3 years (3.2-6.1) (p = 0.05). By Kaplan-Meier survival analysis C homozygotes developed CV significantly earlier than the other cohorts (p = 0.035). At 5 years 100% of C homozygotes had evidence for CV. G homozygotes had a more gradual onset of CV with an approximate 60% prevalence at 5 years. *CC genotype was the most predictive risk factor for CV development (Hazard ratio 4.2 (95% CI 1.3-12.9); p = 0.014). Increasing donor age was also significant (Hazard ratio 1.04 (95% CI, 1.0-1.08); p = 0.023). CONCLUSIONS: Polymorphism at position -174 within the promoter region of the IL-6 gene may be an important risk factor for cardiac transplant related coronary vasculopathy. This may improve patient selection and allow tailored immunosuppressive treatment. PMID- 15896754 TI - Cellular and hemodynamics responses of failing myocardium to continuous flow mechanical circulatory support using the DeBakey-Noon left ventricular assist device: a comparative analysis with pulsatile-type devices. AB - BACKGROUND: An increasing number of continuous flow pumps are currently under clinical studies, however very little data exist on the hemodynamic and cellular responses of the failing heart to continuous flow support. The purpose of this investigation was to characterize the response of the failing myocardium to continuous flow support. METHODS: We compared echocardiographic and cellular markers of failing myocardium at the time of left ventricular assist device (LVAD) implantation and explantation in 20 consecutive patients (12 pulsatile flow [Novacor] and 8 continuous flow [DeBakey-Noon]). RESULTS: The use of mechanical support with both continuous- or pulsatile-type LVADs resulted in a reduction of left ventricular end-diastolic dimension (LVEDD), end-diastolic volume (EDV), end-systolic volume (ESV) and left atrial volume (LAV), as well as a decrease in mitral E/A ratio, tricuspid regurgitation velocity (TRV) and pulmonary valve acceleration time (PVAT). Comparative analyses for patients treated with a continuous- vs pulsatile-type LVAD support showed a greater degree of unloading with the latter type, as shown by the effect on LVEDD (-13.7% vs 33.7%, p = 0.0.004), EDV (-23.5% vs -41.2%, p = 0.015), ESV (-25.6% vs -57.6%, p = 0.001) and LAV (-25.2% vs -40.4%, p = 0.071). The hemodynamic effects of continuous vs pulsatile LVAD support were similar, as shown by their effect on mitral E/A ratio (-23.9% vs -39.9%, p = NS), TRV (-26.4% vs -23.8%, p = NS) and PVAT (28.5% vs 38.5%, p = NS). Only pulsatile support demonstrated a statistically significant percent change in mass (-6.3% vs -20.6%, p = 0.038). Continuous and pulsatile forms of mechanical support demonstrated equivalent reductions in myocardial tumor necrosis factor-alpha (TNF-alpha), total collagen and mycocyte size. CONCLUSIONS: Our findings show that, although there are differences between these 2 devices in magnitude of unloading, both forms of support effectively normalize cellular markers of the failing phenotype. PMID- 15896755 TI - Fate of infants with hypoplastic left heart syndrome listed for cardiac transplantation: a multicenter study. AB - BACKGROUND: Infants with hypoplastic left heart syndrome (HLHS) commonly undergo cardiac transplantation as primary management. METHODS: We examined outcomes of primary transplantation for unpalliated HLHS. We analyzed data from the 20 institutions of the Pediatric Heart Transplant Study Group, from January 1, 1993, through December 31, 1998, using actuarial and parametric survival analysis and competing outcomes analysis. RESULTS: During the 6 years studied, 1,234 patients were listed for cardiac transplantation; 262 patients (21.2%) had unpalliated HLHS. The number (and percentage) of patients with HLHS decreased from 58 (27% of patients listed) in 1993 to 30 (14%) in 1998. Overall, 25% of infants with HLHS died while waiting; primary cause of death was cardiac failure (50%). Of the remaining patients awaiting transplantation, 23 (9%) underwent Norwood/Fontan type surgeries as interim palliation: 52% died. Ultimately, 175 patients underwent cardiac transplantation (67%); 50% received organs by 2 months after listing. Post-transplant actuarial survival was 72% at 5 years, with 76% of deaths (35/46) occurring within 3 months; early mortality was caused primarily by graft failure within the first 30 days after transplantation (in 54%). Among 1 month survivors, survival at 1 and at 5 years was 92% and 85%, respectively. Of the 262 patients listed with unpalliated HLHS, overall survival, taking into account mortality after listing and after transplantation, was 68% at 3 months and 54% at 5 years. CONCLUSIONS: Cardiac transplantation offers good intermediate survival for infants with unpalliated HLHS. PMID- 15896756 TI - Extended ganciclovir prophylaxis in lung transplantation. AB - BACKGROUND: Positive cytomegaloviral status of the donor or of the recipient adversely affects survival and enhances the development of bronchiolitis obliterans syndrome (BOS) in lung transplant recipients. The role of ganciclovir prophylaxis in cytomegalovirus infection in respect to obliterative bronchiolitis or to BOS development is not known. METHODS: From the Papworth transplant database, we identified 146 patients who received organs from cytomegalovirus positive donors. We classified patients into 3 groups as follows: Group 1 consisted of 42 patients who underwent transplantation between 1990 and 1992 when no prophylaxis was given; Group 2 consisted of 49 patients who underwent transplantation between 1992 and 1995 when 4 weeks of IV ganciclovir was given as prophylaxis; and Group 3 consisted of 55 patients who underwent transplantation between 1995 and 1998 when cytomegalovirus prophylaxis consisted of IV (1 week) followed by oral ganciclovir for a total of 3 months. Donor management, recipient management during and after surgery, and pharmacotherapy were uniform during the study period. We used survival and regression methods to compare these groups, adjusting for the transplantation type (single lung, double lung, or heart-lung) and for HLA typing. RESULTS: We found a significant difference among all 3 groups in numbers of cytomegaloviral disease episodes in the 1st year after transplantation. The number of rejection episodes in the 3 groups during the 1st post-transplant year gradually decreased from Group 1 to Group 3. We identified no statistically significant benefit in the time to BOS occurrence or in actuarial survival. CONCLUSION: Extended prophylaxis with IV and oral ganciclovir practically abolishes cytomegaloviral disease and is related to a decreased incidence of rejection episodes. However, ganciclovir prophylaxis is not related to a decreased incidence or progression of BOS or survival. PMID- 15896757 TI - Safety and efficacy of two types of influenza vaccination in heart transplant recipients: a prospective randomised controlled study. AB - BACKGROUND: Influenza may cause severe disease in immunosuppressed patients. Different vaccines have been proved to be efficacious to prevent influenza in tranplant recipients. Since the last five years the addition of adjuvants to improve the immune response to vaccine preparations has been proposed and evaluated. In this study, two antigenically identical vaccines, but different for the presence of adjuvants were randomised among a cohort of heart transplant recipients to evaluate their safety and immunogenicity. METHODS: 58 patients, receiving an heart transplant more than 6 months before, were randomised to receive one shoot vaccination with Fluad (containing the MF59 adjuvant) or Agrippal (no adjuvant added) or to enter the control, not-vaccinated, group. The immune response to influenza was evaluated separately for type A and type B viruses and for the IgG and the IgM antibodies. Patients were clinically evaluated at least monthly up to 6 months. RESULTS: Influenza symptoms were reported by 33% of patients receiving Fluad, 29% of the Agrippal and 63% of the control group. 4 episodes of acute myocardial rejection >/=3A were identified without difference between the three groups. CONCLUSIONS: The superior efficacy of vaccines containing adjuvants was not found and the data clearly confirmed that vaccination against influenza is safe and effective in heart transplant recipients. The use of vaccine containing adjuvant substances do not ameliorate the clinical performance of the immunisation suggesting that less expensive influenza vaccine preparation without adjuvant substances could be equally useful to protect heart transplant recipients. PMID- 15896758 TI - Endothelial activation in the transplanted human heart from organ retrieval to 3 months after transplantation: an observational study. AB - BACKGROUND: Endothelial activation in the donor heart has been described variably after brain death and transplantation. We aimed to characterize the time course of endothelial activation in right ventricle (RV) and left ventricle (LV) during the acute phase of clinical transplantation. METHODS: We studied biopsy specimens from the RVs and the LVs of 40 donor hearts: at initial assessment of the donor, at end-ischemia, and after 10 minutes of reperfusion. We also included follow-up RV biopsy specimens at 1 week, 1 month, and 3 months after surgery. Six of the patients had cystic fibrosis and were domino donors. RESULTS: P-selectin and vascular cell adhesion molecule 1 (VCAM-1), but not E-selectin were up-regulated in brain-dead and in domino donors vs controls. Unused donor hearts (n = 6) had significantly less up-regulation of P-selectin and of VCAM-1. We found no difference between the RV and the LV during surgery, but we did see important time-dependent variations. P-selectin was present in 85% of vessels throughout transplantation and decreased to approximately 60% after transplantation (p < 0.001). We initially detected VCAM-1 in 20% of vessels, which decreased to 5% during storage, then increased to 47% at reperfusion, and gradually decreased thereafter (p < 0.001). E-selectin expression increased progressively from 15% initially to 45% at reperfusion and then decreased after surgery (p = 0.001). Thrombomodulin expression was decreased at baseline, and the decrease was accentuated afterward (p = 0.02). Patients with donor organ failure did not have a specific pattern of endothelial activation. CONCLUSION: Cardiac transplantation is associated with marked endothelial activation, with no difference between the two ventricles. The changes persist in the post-operative period, even in the absence of acute rejection. PMID- 15896759 TI - Long-term preservation using a new apparatus combined with suppression of pro inflammatory cytokines improves donor heart function after transplantation in a canine model. AB - OBJECTIVE: We developed a new apparatus for long-term heart preservation that combines simple immersion with coronary perfusion. In a previous study, we reported that suppression of pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta), improved results after transplantation. In this study, we evaluated whether long-term preservation using our apparatus for continuous coronary perfusion, combined with suppression of pro inflammatory cytokines, improves donor heart function after transplantation in a canine model. METHODS: We used adult mongrel dogs in this study. Coronary vascular beds were washed with University of Wisconsin (UW) solution after arresting hearts with glucose-insulin-potassium solution. The heart was then excised and preserved for 12 hours with a combination of immersion and coronary perfusion using a preservation apparatus. Adult mongrel dogs were divided into 2 groups: the coronary perfusion (CP) group (n = 7) and the FR167653 (FR-CP) group (n = 6). In the CP group, we used a 4 degrees C UW solution for immersion and coronary perfusion. In the FR-CP group, we used a 4 degrees C UW solution supplemented with 20 mg/liter of the anti-inflammatory agent FR167653 for immersion and coronary perfusion. At 2 and at 3 hours after orthotopic transplantation, we compared hemodynamic parameters with pre-operative values in donor animals, with right atrial pressure at 10 mm Hg and with 5 microg/kg/min dopamine infusion. We compared serum concentrations of TNF-alpha from the coronary sinus and compared electron microscopic studies between the 2 groups. RESULTS: Three hours after transplantation, cardiac output (CO), left ventricular pressure (LVP), and -LVdp/dt were significantly greater (p < 0.05) in the FR-CP group than in the CP group (CO, 178% +/- 65% vs 93% +/- 40%; LVP, 115% +/- 22% vs 73% +/-26%; -LVdp/dt, 168% +/- 13% vs 61% +/- 17%, respectively). Electron microscopic studies showed that glycogen was well preserved in the FR-CP group compared with the CP group. Serum concentrations of TNF-alpha were decreased significantly in the FR-CP group compared with the CP group at 3 hours after reperfusion (161 +/- 54 pg/dl vs 642 +/- 636 pg/dl, respectively). CONCLUSION: Hemodynamics after transplantation were significantly better in the FR-CP group than in the CP group. The combined preservation method of continuous perfusion and immersion using our apparatus in conjunction with suppression of pro inflammatory cytokines improves donor heart function after transplantation. PMID- 15896760 TI - Alterations in alpha adrenoreceptor density and localization after mechanical left ventricular unloading with the Jarvik flowmaker left ventricular assist device. AB - Cardiac alpha one adrenoreceptors (alpha(1)AR) are known to mediate positive inotropism in human ventricular myocardium. In the early stages of heart failure, ventricular contractility is maintained by adrenergic stimulation, rennin angiotensin activation, and other neurohormonal and cytokine system responses. As the disease progresses, however, these compensatory mechanisms cease to provide benefit; ventricular dilation and fibrosis occur and cardiac function deteriorates. When ventricular contractility becomes severely depressed and is no longer responsive to inotropic support, insertion of a left ventricular assist device (LVAD) may allow the left ventricle to rest, remodel, and recover some contractile function. Our previous work has demonstrated that the myocardium has the capacity to repair itself during a period of unloading, after which some patients are able to resume a normal lifestyle and no longer need a cardiac transplant. PMID- 15896761 TI - Blood transfers infectious immunologic tolerance in MHC-incompatible heart transplantation in rats. AB - "Infectious tolerance" or inducing immunologic tolerance of infection in allografts is still poorly understood. We investigated whether transfusing blood from LEW.1A rats tolerant of LEW.1W hearts could transmit tolerance to naive LEW.1A rats. In 4 of 6 cases, transfusing blood from tolerant animals was followed by immunologic tolerance of heart transplants from LEW.1W donor rats in LEW.1A recipient animals, whereas transplanting heart grafts that were tolerated in previous transplantations across MHC barriers did not transfer tolerance in major histocompatibility complex (MHC)-incompatible animals. We conclude that in rat heart transplantation, the transfer of immunologic tolerance can be enhanced by transfusing blood from tolerant animals to naive animals before transplantation across MHC barriers. PMID- 15896762 TI - End-stage cardiac failure caused by isolated ventricular non-compaction: cardiac transplantation in a 5-year-old boy. AB - Isolated ventricular non-compaction (IVNC) is a rare congenital cardiomyopathy characterized by arrest in endomyocardial morphogenesis. Presenting features in children include systolic and diastolic ventricular dysfunction, cardiac arrhythmias, and thromboembolism. End-stage cardiac failure necessitating cardiac transplantation is described in adults. We report the unique case of a 5-year-old boy with IVNC and end-stage heart failure who underwent cardiac transplantation. Diagnostic echocardiographic features, gross cardiac morphology, and histopathologic findings are illustrated. PMID- 15896763 TI - West Nile virus encephalitis infection in a heart transplant recipient: a case report. AB - Infection with West Nile virus (WNV) ranges from causing no symptoms to causing non-specific fever, meningitis, meningo-encephalitis and death. Advanced age and immunosuppression are the greatest risk factors for the development of encephalitis or other neurologic manifestations of WNV. We report the first fatal case of primary WNV infection in an orthotopic heart transplant recipient. We reviewed the literature on WNV infection in the general population and in immunocompromised hosts. As WNV becomes more common in the United States, it will become an increasing threat to our transplant population. PMID- 15896764 TI - Pulmonary embolization of microcrystalline cellulose in a lung transplant recipient. AB - Intravenous injection of drugs that contain insoluble foreign material can lead to pulmonary embolization of the material and can have devastating results, including pulmonary hypertension and death. Most cases are detected after the onset of extensive, irreversible damage, precluding potentially life-saving intervention, or are detected at autopsy. We report here a case of microcrystalline cellulose embolization in a lung transplant recipient detected at routine transbronchial biopsy, and we describe the circumstances associated with the development of this condition and its clinical outcome. PMID- 15896765 TI - Use of rituximab to decrease panel-reactive antibodies. AB - Patients with pre-formed antibodies may be at increased risk of rejection after organ transplantation. In this report, we describe the use of rituximab to decrease the percentage of pre-formed antibodies in a pediatric heart transplant recipient. PMID- 15896766 TI - Hemodynamic observations in two pulmonary hypertensive patients changing treatment from inhaled iloprost to the oral endothelin-antagonist bosentan. AB - Two female patients with pulmonary arterial hypertension and a permanently implanted hemodynamic monitor changed treatment from inhaled iloprost to oral bosentan. The hemodynamic changes were seen very early after the first dose of bosentan and there was no need to re-establish inhaled iloprost. PMID- 15896767 TI - Recurrence of pulmonary veno-occlusive disease after heart-lung transplantation. AB - Pulmonary veno-occlusive disease, a rare cause of pulmonary hypertension, is characterized by extensive and diffuse occlusion of pulmonary veins by fibrous tissue. Although the diagnosis can be suspected by the presence of the classic clinical triad of severe pulmonary arterial hypertension, radiographic evidence of pulmonary hypertension and edema, and normal pulmonary artery occlusion pressure, the definitive diagnosis is histopathologic. The prognosis of pulmonary veno-occlusive disease is poor with most described patients dying within 2 years of diagnosis. Although anti-coagulation, oxygen, and vasodilator therapies are effective temporarily, the definitive treatment is lung transplantation. We describe the recurrence of pulmonary veno-occlusive disease at 3 months after heart-lung transplantation in a 26-year-old man. Recurrence after transplantation for this disease has not been reported previously, and lung transplantation was thought to be definitive treatment. With this 1st report of early recurrence of pulmonary veno-occlusive disease after heart-lung transplantation, we believe that extrapulmonary factors may play a role in the pathogenesis of this rare disease. PMID- 15896768 TI - Ascending aortic aneurysm after pediatric heart transplantation: case report of an unusual complication. AB - A 28-month-old boy, weighing 11 kg, with severe dilated cardiomyopathy, was transplanted on December 1995. Hypertension and supraventricular tachycardia were detected in the immediate post-operative period, with favorable outcome. After 5 months of clinically asymptomatic follow-up, a dilation in the ascending aorta was observed on routine echocardiogram. Nuclear magnetic resonance imaging (NMRI) confirmed an ascending aortic aneurysm, with a diameter of 38 mm. An operation was performed, a bovine pericardium patch was sutured with reconstruction of the aortic wall, excluding the aneurysm. Good recovery was obtained and the child was discharged on Day 7 postoperatively. A post-operative echocardiogram showed absence of the aortic aneurysm and good surgical results. Another NMRI was done 5 months later, showing an intact ascending aorta. After 64 months, the patients clinical condition was confirmed as normal by echocardiogram. Surgical treatment was successful and the positive results have been maintained. PMID- 15896769 TI - Cellulitis caused by Cryptococcus neoformans in a lung transplant recipient. PMID- 15896770 TI - Deep vein thrombosis after transfemoral endomyocardial biopsy in cardiac transplant recipients. PMID- 15896771 TI - The role of laminin-integrin signaling in triggering MB formation. An in vivo and in vitro study. AB - It is still unclear as to how hepatocytes perceive external factors and transduce the signals which initiate MB formation. To investigate this phenomenon, the model of MB formation in liver in vivo and in primary culture of hepatocytes derived from drug-primed mice was used. Control mice were fed the control diet (group 1). MBs were induced in the livers of mice fed diethyl-1, 4-dihydro-2, 4, 6-trimethyl-3, 5-pyridinedicarboxylate (DDC) for 10 weeks (group 2). The induced MBs completely disappeared after the withdrawal of DDC for 4 weeks (group 3). Newly formed MBs were numerous after DDC was refed for 1 week (group 4). Relative mRNA abundance was determined by quantitative real-time RT-PCR in the liver from the mice. The expression of integrin alpha(6) and beta(2) was significantly increased in the livers of DDC-treated (group 2) and drug refed mice (group 4), when compared with the livers from controls (group 1) and DDC-withdrawn (group 3) mice. The increased mRNA of these two integrin genes was associated with the increased expression of laminin (a ligand for integrin alpha(6)beta(1) and alpha(6)beta(4)), Icam1 (a ligand of alphaLbeta2), Src, MEKK1, and ERK1. Primary cultures of isolated DDC-primed hepatocytes (group 4 mice were withdrawn from DDC CMZ for 4-6 weeks) produced significantly more MBs on laminin-coated coverslips compared with plastic uncoated, fibronectin-, collagen-, or fibrinogen-coated coverslips. U0126, an inhibitor of MEK1 protein, significantly reduced the phosphorylated forms of ERK1/2 and MB formation in vitro. In conclusion, the current study revealed an association between MB formation and integrin-mediated signaling in vivo. The data indicate that laminin-integrin signaling which activates ERK, triggered MB formation in vitro, and an inhibitor of the signaling cascade reduced MB formation. PMID- 15896772 TI - Prevalence of the Factor V G1691A and the Factor II/prothrombin G20210A gene polymorphisms among Tamilians. AB - We have investigated the prevalence of the Factor II G20210A and Factor V G1691A single nucleotide polymorphisms (SNPs) in a South Indian-Tamil Nadu population. The SNP genotyping was performed using a polymerase chain reaction (PCR)/restriction fragment length polymorphism analysis and by a recently FDA approved LightCycler real-time PCR assay. Of 72 samples that were genotyped, 4 (5.5%) patients were heterozygous for the Factor V SNP and no homozygous mutant patients were identified. None of the patients were shown to be either heterozygous or homozygous mutant for the Factor II SNP. All samples showed 100% concordance between the PCR/RFLP assay and the LightCycler assay. While this study identified the prevalence of the Factor V SNP to be similar to that of other reported populations, the absence of the Factor II allele is consistent with reports in more isolated populations. In addition, the results of this study do not support a role for these SNPs in acute myocardial infarction in the Tamilian population. PMID- 15896773 TI - The HIN-200 family: more than interferon-inducible genes? AB - The HIN-200 family was initially grouped together based on their hemopoietic expression, interferon-inducibility, nuclear localization, and characteristic 200 amino-acid domains. In this review, we performed a comprehensive search of genome databases and determined the location of previously characterized and predicted genes within the human, mouse, and rat HIN-200 loci. Several novel proteins were predicted in the mouse and rat. We also discuss recent advances in our understanding of this family of proteins and highlight the most important findings. In addition to a role in interferon biology, there is now good evidence supporting a role for these proteins as regulators of cell proliferation and differentiation. The activity of HIN-200 proteins is not restricted to the hemopoietic system as they are expressed and can function in a variety of other cells and tissues. The importance of HIN-200 proteins in disease now is beginning to be understood as they appear to be involved in autoimmunity and may act as tumor suppressor proteins. PMID- 15896774 TI - In vivo dynamics and kinetics of pKi-67: transition from a mobile to an immobile form at the onset of anaphase. AB - A cell proliferation marker protein, pKi-67, distributes to the chromosome periphery during mitosis and nucleolar heterochromatin in the interphase. We report here on the structural domains of pKi-67 that are required for its correct distribution. While both the LR domain and the conserved domain were involved in localization to the nucleolar heterochromatin, both the LR domain and the Ki-67 repeat domain were required for its distribution to the mitotic chromosome periphery. Using in vivo time-lapse microscopy, GFP-pKi-67 was dynamically tracked from the mitotic chromosome periphery to reforming nucleoli via prenucleolar bodies (PNBs). The signals in PNBs then moved towards and fused into the reforming nucleoli with a thin string-like fluorescence during early G1 phase. An analysis of the in vivo kinetics of pKi-67 using photobleaching indicated that the association of pKi-67 with chromatin was progressively altered from "loose" to "tight" after the onset of anaphase. These findings indicate that pKi-67 dynamically alters the nature of the interaction with chromatin structure during the cell cycle, which is closely related to the reformation process of the interphase nucleolar chromatin. PMID- 15896775 TI - Activation of c-Jun N-terminal kinase (JNK) pathway by HSV-1 immediate early protein ICP0. AB - The immediate early protein ICP0 encoded by herpes simplex virus 1 (HSV-1) is believed to activate transcription and consequently productive infection. The precise mechanisms of ICP0-mediated transactivation are under intensive study. Here, we demonstrate that ICP0 can strongly activate AP-1 responsive genes specifically. This activation is inhibited by c-Jun (S73A), c-Jun (S63/73A), TAK1 (K63W), but not by p38 (AF), ERK1 (K71R), ERK2 (K52R) and TRAF6 (C85A/H87A). We further investigate the relevancy of ERK, JNK and p38 MAPK pathways using their respective inhibitors PD98059, SP600125 and SB202190. Only SP600125 significantly attenuates the AP-1 responsive gene activation by ICP0. Consistent with these, the JNK is remarkably activated in response to ICP0, and this JNK activation is shown to be significantly attenuated by TAK1 (K63W). It turns out that ICP0 interacts specifically with TAK1 and stimulates its kinase activity. These findings reveal a new molecular mechanism ICP0 explores to regulate gene expression. PMID- 15896776 TI - Co-expression of SOX9 and SOX10 during melanocytic differentiation in vitro. AB - Investigations into pigment cell biology have relied on the ability to culture both murine and human melanocytes, numerous melanoma cell lines and more recently, murine and human melanoblasts. Melanoblast culture requires medium supplemented with a range of growth factors including Stem Cell Factor, Endothelin-3 and Fibroblast Growth Factor-2, withdrawal of which causes the cells to differentiate into melanocytes. Using the human melanoblast culture system, we have now examined the expression and/or DNA binding activity of several transcription factors implicated in melanocytic development and differentiation. Of these, the POU domain factor BRN2 and the SOX family member SOX10 are both highly expressed in unpigmented melanocyte precursors but are down-regulated upon differentiation. In contrast, the expression levels of the previously described MITF and PAX3 transcription factors remain relatively constant during the melanoblast-melanocyte transition. Moreover, BRN2 ablated melanoma cells lack expression of SOX10 and MITF but retain PAX3. A novel finding implicates a second SOX protein, SOX9, as a potential melanogenic transcriptional regulator, as its expression level is increased following the down-regulation of BRN2 and SOX10 in differentiated melanoblasts. Our results suggest that a complex network of transcription factor interactions requiring proper temporal coordination is necessary for acquisition and maintenance of the melanocytic phenotype. PMID- 15896777 TI - The constitutively active N111G-AT1 receptor for angiotensin II modifies the morphology and cytoskeletal organization of HEK-293 cells. AB - The expression of a constitutively active G protein-coupled receptor is expected to trigger diverse cellular changes ranging from normal to adaptive responses. We report that confluent HEK-293 cells stably expressing the constitutively active mutant N111G-AT1 receptor for angiotensin II spontaneously exhibited dramatic morphological changes and cytoskeletal reorganization. Phase-contrast microscopy revealed that these cells formed a dense monolayer, whereas cells expressing the WT-AT1 receptor displayed large intercellular spaces and numerous filopodia. Confocal microscopy revealed an elaborate web of polymerized actin at the apical and basolateral surfaces of cells expressing the N111G-AT1 receptor. Interestingly, these phenotypic changes were prevented by culturing the cells in the presence of the inverse agonist EXP3174. Similar morphologic rearrangements and de novo polymerized actin structures were found in Ang II-stimulated cells expressing the WT-AT1 receptor. We further showed that AT1 receptor-induced cell cell contact formation did not require an increase in intracellular Ca2+ concentration or the activity of protein kinase C. However, pretreatment with Y 27632 revealed that Rho-kinase activity was required for cell-cell contact formation upon AT1 receptor activation. These observations demonstrate that the expression of the constitutively active mutant N111G-AT1 receptor had a significant impact on the morphology and cytoskeletal organization of HEK-293 cells, possibly via a mechanism involving the activity of Rho-kinase. PMID- 15896778 TI - D-threo-tetrahydrobiopterin is synthesized via 1'-oxo-2'-D-hydroxypropyl tetrahydropterin in Dictyostelium discoideum Ax2. AB - The biosynthesis of D-threo-tetrahydrobiopterin (DH4, tetrahydrodictyopterin) in Dictyostelium discoideum Ax2 was investigated through the mutant disrupted in the gene encoding sepiapterin reductase (SR) by insertional inactivation. The mutant cells, being completely devoid of SR protein, showed 18.1% of L-erythro tetrahydrobiopterin (BH4) and 0.6% of DH4 productions in the wild type cells. The mutant cells were also identified to excrete D- and L-sepiapterin, which were presumed to originate from intracellular 1'-oxo-2'-D-hydroxypropyl- and 1'-oxo-2' L-hydroxypropyl-tetrahydropterin (H4-pterin), respectively. Furthermore, in a coupled assay with Dictyostelium SR, the mutant cell extract exhibited a novel enzyme activity converting 6-pyruvoyltetrahydropterin to 1'-oxo-2'-D hydroxypropyl-H4-pterin. These results are clear demonstration of the in vivo synthesis of DH4 via 1'-oxo-2'-D-hydroxypropyl-H4-pterin as well as an alternative synthesis of BH4 and DH4 in the complete absence of SR. PMID- 15896779 TI - Phosphorylation of a tyrosine at the N-terminus regulates the surface expression of GIRK5 homomultimers. AB - The G protein-coupled inwardly rectifying GIRK5 and Delta5GIRK5 splicing variants do not express functional potassium channels. In contrast, Delta25GIRK5 forms functional homomultimers in Xenopus laevis oocytes. A tyrosine is present at the N-term of the non-functional isoforms. We studied the effect of endogenous tyrosine phosphorylation on the GIRK5 surface and functional expression. Unlike wild type channels, GIRK5Y16A and Delta5GIRK5Y16A mutants displayed inwardly rectifying currents and inhibitors of Src tyrosine kinase promoted the traffiking of GIRK5 to the cell surface. This is the first evidence that endogenous phosphorylation of a tyrosine residue in a GIRK channel inhibits its surface expression. PMID- 15896780 TI - Differential interactions of the homeodomain-interacting protein kinase 2 (HIPK2) by phosphorylation-dependent sumoylation. AB - Homeodomain-interacting protein kinase 2 (HIPK2) interacts with and phosphorylates various transcription factors that are critical regulators of cell fate decisions and apoptosis during development. Here we show that lysine 25 of HIPK2 is the major sumoylation site, both in vitro and in vivo, and that the sumoylation of this site occurs in a phosphorylation-dependent manner. This became clear with the finding that kinase-dead HIPK2 (K221R) could not be efficiently sumoylated in vitro. The sumoylation of HIPK2 resulted in the disruption of its interaction with a Groucho corepressor. Consequently, sumoylation inhibited the regulatory activity of HIPK2 on the Groucho-mediated repression of transcription, whereas not on p53-mediated transactivation. These results suggest that phosphorylation-dependent sumoylation enables HIPK2 to drive different target gene transcription by means of differential interactions with its binding partners. PMID- 15896781 TI - Heterogeneities in EGF receptor density at the cell surface can lead to concave up scatchard plot of EGF binding. AB - The mechanism responsible for the concave up nature of the Scatchard plot of epidermal growth factor (EGF) binding on EGF receptor (EGFR) has been a controversial issue for more than a decade. Past efforts to mechanistically simulate the concave up nature of the Scatchard plot of EGF binding have shown that negative cooperativity in EGF binding on an EGFR dimer or inclusion of some external site or binding event can describe this behavior. However, herein we show that heterogeneity in the density of EGFR due to localization in certain regions of the plasma membrane, which has been experimentally reported, can also lead to concave up shape of the Scatchard plot of the EGF binding on EGFR. PMID- 15896782 TI - Constitutive dimerization of human serotonin 5-HT4 receptors in living cells. AB - Serotonin 5-HT4 receptor isoforms are G protein-coupled receptors (GPCRs) with distinct pharmacological properties and may represent a valuable target for the treatment of many human disorders. Here, we have explored the process of dimerization of human 5-HT4 receptor (h5-HT4R) by means of co-immunoprecipitation and bioluminescence resonance energy transfer (BRET). Constitutive h5-HT4(d)R dimer was observed in living cells and membrane preparation of CHO and HEK293 cells. 5-HT4R ligands did not influence the constitutive energy transfer of the h5-HT4(d)R splice variant in intact cells and isolated plasma membranes. In addition, we found that h5-HT4(d)R and h5-HT4(g)R which structurally differ in the length of their C-terminal tails were able to form constitutive heterodimers independently of their activation state. Finally, we found that coexpression of h5-HT4R and beta2-adrenergic receptor (beta2AR) led to their heterodimerization. Given the large number of h5-HT4R isoforms which are coexpressed in a same tissue, our results points out the complexity by which this 5-HTR sub-type mediates its biological effects. PMID- 15896784 TI - Effects of disialoganglioside GD3 on the mitochondrial membrane potential. AB - GD3 is an intracellular mediator of apoptotic signaling. Although GD3 is known to directly act on mitochondria, the dynamic responses of individual mitochondria to GD3 remain to be elucidated. In the current study, the membrane potential of single mitochondria is observed in the presence of GD3 or its analogues. Here, we report that (1) GD3 specifically induces gradual depolarizations of the inner membrane by a mechanism that differs from the permeability transition, and (2) the GD3-induced depolarizations are suppressed by cyclosporin A. These results suggest that GD3 depolarizes mitochondria by a mechanism distinct from but relevant to the permeability transition. PMID- 15896785 TI - Knock-down of LAR protein tyrosine phosphatase induces insulin resistance. AB - To test the role of the leukocyte common antigen-related protein tyrosine phosphatase (LAR) as a regulator of insulin receptor (IR) signalling, an siRNA probe against LAR was developed. Knock-down of LAR induced post-receptor insulin resistance with the insulin-induced activation of PKB/Akt and MAP kinases markedly inhibited. The phosphorylation and dephosphorylation of the IR and insulin receptor substrate (IRS) proteins were unaffected by LAR knock-down. These results identify LAR as a crucial regulator of the sensitivity of two key insulin signalling pathways to insulin. Moreover, the siRNA probe provides a molecular tool of general applicability for further dissecting the precise targets and roles of LAR. PMID- 15896786 TI - Proteins and peptides. Proceedings of the joint 30th FEBS Congress and the 9th IUBMB Conference. July 2-7, 2005. Budapest, Hungary. PMID- 15896787 TI - CysMap and CysJoin: database and tools for protein disulphides localisation. AB - We have developed a computer program able to make user-customised databases derived from the public PIR non-redundant reference protein database. When the database of interest has been created, the user will generate the map of all the possible linear peptides containing one and two cysteines for each protein and combine them to calculate the mass of all the possible clusters of linear peptides linked by a disulphide bridge with a cysteine pair. It is also possible to create selected maps corresponding to peptides formed by the action of specific proteases. In this way, mass spectrometric data obtained from the hydrolysis of proteins of unknown sequence can be related to that contained in the database for quick disulphide assignment and protein identification. To confirm signal attribution, the program will also furnish the expected mass of cluster peptides after performing a cycle of Edman degradation. The utility of the program is discussed and examples of application are given. PMID- 15896788 TI - Ammonia displaces methanol bound to the water oxidizing complex of photosystem II in the S2 state. AB - Ammonia and methanol both bind to the water oxidising complex of photosystem II during its turnover, possibly at sites where water binds during the normal water oxidation process. We have investigated the interaction between these two water analogues at the S2 state of the water oxidising cycle using electron magnetic resonance techniques. We find evidence that ammonia displaces methanol from its binding site. PMID- 15896789 TI - Role of Nrf2 signaling in regulation of antioxidants and phase 2 enzymes in cardiac fibroblasts: protection against reactive oxygen and nitrogen species induced cell injury. AB - Understanding the molecular pathway(s) of antioxidant gene regulation is of crucial importance for developing antioxidant-inducing agents for the intervention of oxidative cardiac disorders. Accordingly, this study was undertaken to determine the role of Nrf2 signaling in the basal expression as well as the chemical inducibility of endogenous antioxidants and phase 2 enzymes in cardiac fibroblasts. The basal expression of a scope of key cellular antioxidants and phase 2 enzymes was significantly lower in cardiac fibroblasts derived from Nrf2-/- mice than those from wild type control. These include catalase, reduced glutathione (GSH), glutathione reductase (GR), GSH S transferase (GST), and NAD(P)H:quinone oxidoreductase-1 (NQO1). Incubation of Nrf2+/+ cardiac fibroblasts with 3H-1,2-dithiole-3-thione (D3T) led to a significant induction of superoxide dismutase (SOD), catalase, GSH, GR, glutathione peroxidase (GPx), GST, and NQO1. The inducibility of SOD, catalase, GSH, GR, GST, and NQO1, but not GPx by D3T was completely abolished in Nrf2-/- cells. The Nrf2-/- cardiac fibroblasts were much more sensitive to reactive oxygen and nitrogen species-mediated cytotoxicity. Upregulation of antioxidants and phase 2 enzymes by D3T in Nrf2+/+ cardiac fibroblasts resulted in a dramatically increased resistance to the above species-induced cytotoxicity. In contrast, D3T-treatment of the Nrf2-/- cells only provided a slight cytoprotection. Taken together, this study demonstrates for the first time that Nrf2 is critically involved in the regulation of the basal expression and chemical induction of a number of antioxidants and phase 2 enzymes in cardiac fibroblasts, and is an important factor in controlling cardiac cellular susceptibility to reactive oxygen and nitrogen species-induced cytotoxicity. PMID- 15896790 TI - Photothermal studies of CO photodissociation from mixed valence Escherichia coli cytochrome bo3. AB - Here, we report the volume and enthalpy changes accompanying CO photodissociation from the mixed valence form of cytochrome bo3 oxidase from Escherichia coli. The results of photoacoustic calorimetry indicate two kinetic phases with distinct volume and enthalpy changes accompanying CO photodissociation from heme o3 and its transfer to CuB. The first phase occurring on a timescale of <50 ns is characterized by a volume decrease of -1.3+/-0.3 mL mol-1 and enthalpy change of 32+/-1.6 kcal mol-1. Subsequently, a volume increase of 2.9 mL mol-1 with an enthalpy change of -5.3+/-2.5 kcal mol-1 is observed with the lifetime of approximately 250 ns (this phase has not been detected in previous optical studies). These volume and enthalpy changes differ from the volume and enthalpy changes observed for CO dissociation from fully reduced cytochrome bo3 oxidase indicating that the heme o3/CuB active site dynamics are affected by the redox state of heme b. PMID- 15896791 TI - Robustness of the p53 network and biological hackers. AB - The p53 protein interaction network is crucial in regulating the metazoan cell cycle and apoptosis. Here, the robustness of the p53 network is studied by analyzing its degeneration under two modes of attack. Linear Programming is used to calculate average path lengths among proteins and the network diameter as measures of functionality. The p53 network is found to be robust to random loss of nodes, but vulnerable to a targeted attack against its hubs, as a result of its architecture. The significance of the results is considered with respect to mutational knockouts of proteins and the directed attacks mounted by tumour inducing viruses. PMID- 15896793 TI - House sparrows (Passer domesticus) adjust their social status position to their physiological costs. AB - For group-living animals, the maintenance of a position in the social hierarchy may be associated with physiological costs such as increased stress and energy expenditure or suppressed immune functions. In this study, we experimentally manipulated the social status of house sparrows so that each bird experienced two social environments in random sequence: being dominant and subordinate. For 14 males, we investigated how corticosterone concentrations, energy expenditure and immune functions were affected by these changes in social status position. We found that the cost of maintaining a social status position differed between individuals and were related to individual body size. Birds with small body size had increased costs in terms of increased stress responses and reduced cell mediated immune responses while being experimentally kept as dominants, while birds with large body size had increased costs while they were subordinates. We also found that birds with increased energetic and immunological costs as dominants obtained a low status position in the large group, while birds with increased costs as subordinates obtained a high status position in the large group. In summary, we found that the costs associated with the maintenance of social status position differed between individuals and was related to the individuals' body size. Furthermore, in a large group, individuals maintained a social status position that minimized energetic and immunological costs. PMID- 15896794 TI - Serotonin, but not melatonin, plays a role in shaping dominant-subordinate relationships and aggression in rainbow trout. AB - The aim of this study was to clarify to what extent the effects of elevated dietary L-tryptophan (Trp) on aggressive behavior and stress responsiveness in rainbow trout are mediated by circulating melatonin and central serotonin (5-HT), respectively. Isolated rainbow trout were paired for 1h a day for 7 days in order to create fish with experience of being dominant and subordinate. Following this week, the fish were tested for aggressive behavior using a resident-intruder test after which they were subjected to one of four treatments: (1) tryptophan, (2) the selective serotonin reuptake inhibitor (SSRI) citalopram, (3) melatonin, and (4) no treatment (controls). After 7 days of treatment, the fish were subjected to a second resident-intruder test. Trp-supplemented feed resulted in a suppression of aggressive behavior in fish with experience of being dominant. Moreover, fish fed Trp-supplemented feed, regardless of social experience, also displayed lower plasma cortisol levels than controls. These effects of elevated dietary Trp were closely mimicked by citalopram treatment, whereas exogenous melatonin had no effect on either aggressive behavior or plasma cortisol. Thus, the effect of elevated dietary Trp on aggressive behavior and stress responses does not appear to be mediated by melatonin even though elevated dietary intake of Trp resulted in an increase in plasma melatonin concentrations. PMID- 15896792 TI - Recent advances in behavioral neuroendocrinology: insights from studies on birds. AB - Ever since investigations in the field of behavioral endocrinology were hatched with experiments on roosters, birds have provided original insights into issues of fundamental importance for all vertebrate groups. Here we focus on more recent advances that continue this tradition, including (1) environmental regulation of neuroendocrine and behavioral systems, (2) steroidogenic enzyme functions that are related to intracrine processes and de novo production of neurosteroids, and (3) hormonal regulation of neuroplasticity. We also review recent findings on the anatomical and functional organization of steroid-sensitive circuits in the basal forebrain and midbrain. A burgeoning body of data now demonstrates that these circuits comprise an evolutionarily conserved network, thus numerous novel insights obtained from birds can be used (in a relatively straightforward manner) to generate predictions for other taxa as well. We close by using birdsong as an example that links these areas together, thereby highlighting the exceptional opportunities that birds offer for integrative studies of behavioral neuroendocrinology and behavioral biology in general. PMID- 15896795 TI - Task-specific recruitment of motor units for vibration damping. AB - Vibrations occur within the soft tissues of the lower extremities due to the heel strike impact during walking. Increases in muscle activity in the lower extremities result in increased damping to reduce this vibration. The myoelectric intensity spectra were compared using principal component analysis from the tibialis anterior and lateral gastrocnemius of 40 subjects walking with different shoe conditions. The soft insert condition resulted in a significant, simultaneous increase in muscle activity with a shift to higher myoelectric frequencies in the period 0-60 ms after heel-strike which is the period when the greater vibration damping occurred. These increases in myoelectric frequency match the spectral patterns which indicate increases in recruitment of faster motor units. It is concluded that fast motor units are recruited during the task of damping the soft-tissue resonance that occurs following heel-strike. PMID- 15896796 TI - In vivo micro particle image velocimetry measurements of blood-plasma in the embryonic avian heart. AB - The measurement of blood-plasma velocity distributions with spatial and temporal resolution in vivo is inevitable for the determination of shear stress distributions in complex geometries at unsteady flow conditions like in the beating heart. A non-intrusive, whole-field velocity measurement technique is required that is capable of measuring instantaneous flow fields at sub-millimeter scales in highly unsteady flows. Micro particle image velocimetry (muPIV) meets these demands, but requires special consideration and methodologies in order to be utilized for in vivo studies in medical and biological research. We adapt muPIV to measure the blood-plasma velocity in the beating heart of a chicken embryo. In the current work, bio-inert, fluorescent liposomes with a nominal diameter of 400 nm are added to the flow as a tracer. Because of their small dimension and neutral buoyancy the liposomes closely follow the movement of the blood-plasma and allow the determination of the velocity gradient close to the wall. The measurements quantitatively resolve the velocity distribution in the developing ventricle and atrium of the embryo at nine different stages within the cardiac cycle. Up to 400 velocity vectors per measurement give detailed insight into the fluid dynamics of the primitive beating heart. A rapid peristaltic contraction accelerates the flow to peak velocities of 26 mm/s, with the velocity distribution showing a distinct asymmetrical profile in the highly curved section of the outflow tract. In relation to earlier published gene-expression experiments, the results underline the significance of fluid forces for embryonic cardiogenesis. In general, the measurements demonstrate that muPIV has the potential to develop into a general tool for instationary flow conditions in complex flow geometries encountered in cardiovascular research. PMID- 15896797 TI - Replacement of the membrane proximal region of I-A(d) MHC class II molecule with I-E-derived sequences promotes production of an active and stable soluble heterodimer without altering peptide-binding specificity. AB - The MHC class II molecule I-A is the murine homologue of HLA-DQ in humans. The I A and DQ heterodimers display considerable heterodimer instability compared with their I-E and HLA-DR counterparts. This isotype-specific behavior makes the production of soluble I-A and DQ molecules very difficult. We have developed a strategy for production of soluble I-A(d) molecules involving expression of I A(d) as a glycosil phosphatidyl inositol (PI) anchored chimera in Chinese Hamster Ovary (CHO) cells. The regions comprising the membrane proximal segments of I A(d) alpha and beta chains were substituted for the corresponding regions of I-E, and the derived constructs were expressed in CHO cells. Procedures for purification of the soluble class II molecules were optimized and the WT and chimeric molecule were compared for structure, biochemical stability and functionality. Our analysis revealed that the substitutions in the membrane proximal domains improved cell surface expression and thermal stability of I-A(d) without altering the peptide binding specificity of the class II molecule. The results suggest that similar strategies could be used to increase the stability of other unstable class II molecules for in vitro studies. PMID- 15896798 TI - Novel antibody tags from the rat lysosomal protein RT1.DM for immunodetection of recombinant proteins. AB - Previously, two mouse monoclonal antibodies (12B8 and 6D4) were raised against the alpha- and beta-subunits of the rat non-classical MHC class-II heterodimer RT1.DM. Here, I describe the epitope mapping of mAb 12B8 to amino acids alpha3-14 (EASPQAWWDESQ) and mAb 6D4 to amino acids beta35-44 (WDPEEGQIVP). Epitope mapping was conducted by preparing fusion proteins between the alpha and beta chain of RT1.DM for Western detection with mAb 12B8 and 6D4. By mutating non-conserved amino acids of the human orthologue of RT1.DM, the rat epitopes were introduced, thereby making the alpha and beta polypeptides sensitive for mAb 12B8 and 6D4 detection. The epitopes, designated as 12B8 and 6D4, were tested for protein tagging. They were appended to the N- or C-terminus of four human proteins, the tumour suppressor protein VHL (von Hippel-Lindau), SUMO4, MHC class-II DQbeta and -DPbeta for expression in mammalian cells. Western detection, immunoprecipitation and localisation of the tagged proteins were successfully demonstrated. Thus, the 12B8 and 6D4 epitope tag can be universally used for the immunodetection of recombinant proteins and to study protein-protein interactions. PMID- 15896799 TI - Removal of therapeutic anti-lymphocyte antibodies from human sera prior to anti human leukocyte antibody testing. AB - Both monoclonal (e.g. Orthoclone (OKT3), rituximab) and polyclonal (e.g. ATGAM, Thymoglobulin (Thymo)) anti-lymphocyte Abs (ALAs) are used extensively in organ transplantation for immunosuppression induction, desensitization, and treatment of acute rejection. ALAs often interfere with post transplant immunologic monitoring. We describe a method that uses magnetic beads to selectively remove ALAs from patient serum. Rabbit anti-mouse Fc-specific (180 mug), or rabbit anti mouse Fab-specific (180 microg), or rabbit anti-horse heavy and light chain specific and rabbit anti-horse F(ab')2 (200 microg) (Jackson Immunoresearch) was adsorbed to 6.7 x 10(8) Dynabeads M-280 conjugated with sheep anti-rabbit IgG (Dynal Biotech). Fifty microliters of normal human serum (NHS) with 2 microg/ml of OKT3 or 100 microg/ml ATGAM, Thymo, or rituximab were incubated with conjugated beads for several incubations. NHS containing ALAs before and after treatment by the protocol were incubated with human lymphocytes and labeled with FITC-antibody to immunoglobulin of the species used to produce the particular ALA. Residual ALA was determined using flow cytometry. Average median channel for serum with or without ALA was 11.1 and 0.120, respectively for OKT3; 64.4 and 0.344 for ATGAM; 108.5 and 0.200 for Thymo; and 1022.5 and 11.4 for rituximab. Treatment lowered the median channel for serum with OKT3 to 0.103, 0.309 for ATGAM, 0.199 for Thymo, and 12.1 for rituximab. ALAs can be effectively removed from serum by the use of magnetic beads conjugated with Ab specific for ALA thereby permitting immunologic monitoring without interference. PMID- 15896800 TI - Simultaneous detection of multiple cytokines and chemokines from nonhuman primates using luminex technology. AB - Cytokines and chemokines are soluble mediators of the immune system that play a crucial role in intercellular signaling, and in the recruitment of cells to inflammation sites. Identification of these molecules in nonhuman primates (NHP) is crucial for the understanding of complex physiological and pathological mechanisms that occur in these species, and to demonstrate whether these mechanisms function similarly in humans. The Luminex100 system is a bench-top flow cytometer that allows the user to perform up to 100 tests simultaneously in a single tube. Recently, a significant number of commercial vendors have developed kits for the simultaneous detection of multiple cytokines and chemokines of human origin with the Luminex system. These kits were tested for their capacity to recognize chemokines and cytokines of nonhuman primate origin. ELISA and ELISPOT assays were also adapted to the Luminex format, and novel assays based on new combinations of antibodies were developed. PBMC were isolated from blood from chimpanzees, rhesus macaques, baboons, cynomolgus macaques, pig tailed macaques, and African green monkeys; these cells were stimulated in vitro and culture supernatants were used for the determination of cytokines and chemokines. Crossreactivity tables were prepared based on the ability of the reagents to detect cytokines and chemokines in NHP samples with similar intensity to the ones observed in human samples. By mixing commercially available reagents and newly developed ones, a combination has been created that allows for the detection of 20 NHP chemokines and cytokines in a single sample, including G-CSF, GM-CSF, IFN-gamma, IL-1beta, IL-1Ra, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 (p40), IL-17, IL-18, MCP-1, MIP-1alpha, MIP-1beta, RANTES, TNF-alpha, and TNF beta. These reagents may become a very useful resource for scientists working with these NHP species, which are relevant pre-clinical models for human diseases and transplantation because they approximate humans in physiology and genetics more closely than any other animal. PMID- 15896801 TI - Improved multiplex immunoassay performance in human plasma and synovial fluid following removal of interfering heterophilic antibodies. AB - Cytokines, chemokines and soluble adhesion molecules interact in a complex network within the immune system. Fingerprinting of these proteins may allow the use of these proteins as biomarkers for identification of disease, disease subtyping and monitoring therapeutic interventions. We developed a multiplex immunoassay (MIA) for the detection of 30 proteins in a variety of human body fluids such as plasma and synovial fluid (SF). The measurement of these proteins is hampered by the presence of human (auto-) antibodies, which can cause non specific binding. We have validated a novel approach for the removal of interfering immunoglobulins using pre-absorption with protein-L. Interfering (auto-) antibodies, such as rheumatoid factor (RF), were removed using three methods; polyethylene glycol (PEG) precipitation, pre-absorption with human gamma globulin or pre-absorption with protein-L. A significant decrease of RF was observed after a 2 h incubation with protein-L. RF IgM levels were reduced by 89% whereas total IgM, IgG and IgA levels were reduced by 60%. Residual immunoglobulins were blocked with rodent serum and did not interfere with the multiplex immunoassay. Comparing the MIA with a conventional enzyme-linked immunosorbent assay (ELISA) using a panel of spiked plasma samples resulted in correlation coefficients for all mediators between R2 = 0.88 and R2 = 0.99. Intra assay variance was less than 10% whereas inter-assay variance ranged between 6% and 16%. Pathological samples with heterophilic antibodies hamper immunoassays such as ELISA and MIA. We show that pre-absorption with protein-L is a powerful tool for removal of interfering immunoglobulins from human bodily fluids to be used in immunoassays for studying changes in protein patterns. PMID- 15896802 TI - A method to generate antigen-specific mAb capable of staining formalin-fixed, paraffin-embedded tissue sections. AB - Abnormalities in HLA class I antigen expression are frequently found in malignant tumors. Their potential role in the clinical course of the disease and in the outcome of T cell-based immunotherapy has stimulated interest in the characterization of the molecular mechanisms underlying HLA class I antigen abnormalities in malignant cells. Multiple mechanisms have been identified. Among them are abnormalities in antigen processing machinery (APM) component expression. In spite of this information, APM component expression in malignant lesions has been investigated only to a limited extent because of the lack of availability, for most APM components, of monoclonal antibodies (mAb) which stain formalin-fixed, paraffin-embedded tissues. The latter are the substrate of choice in immunohistochemical (IHC) reactions. To overcome this limitation, we have developed a simple and reproducible method to generate APM component-specific mAb which stain formalin-fixed, paraffin-embedded tissue sections. This method involves five steps: (i) immunogenic amino acid sequences, which display low homology with their mouse counterparts when possible, are identified in APM components and utilized to synthesize peptides; (ii) BALB/c mice are immunized with keyhole limpet hemocyanin (KLH)-conjugated synthetic peptides and with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)-purified recombinant APM component proteins; (iii) immunized mice, which develop high titer APM component-specific antibodies, are utilized to generate hybridomas which are screened for APM component-specific antibody production by Western blotting assays, with lymphoid cell lysates; (iv) identified APM component specific mAb are characterized in their specificity and in their reactivity with permeabilized cells in ELISA and/or flow cytometry; and (v) mAb, with the appropriate reactivity pattern, are tested in IHC reactions with formalin-fixed, paraffin-embedded tissue sections. The use of the methodology we have developed resulted in the generation of a panel of APM component-specific mAb capable of staining formalin-fixed, paraffin-embedded tissue sections in IHC reactions. These reagents will facilitate the analysis of APM component expression in tissues under physiological and pathological conditions. In addition, the methodology we have described is likely to be applicable to other antigenic systems to develop mAb capable of detecting protein components of interest in formalin-fixed, paraffin-embedded tissue sections. PMID- 15896803 TI - Influence of ions, hydration, and the transcriptional inhibitor P4N on the conformations of the Sp1 binding site. AB - Three crystal structures containing the entire Sp1 consensus sequence d(GGGGCGGGG) with two or three additional base-pairs on either the 5' or 3' ends and overhangs have been determined. Despite the different lengths of DNA in the pseudo-dodecamers and pseudo-tridecamer, all three structures form A-DNA duplexes that share a common set of crystal contacts, including a T*(G.C) base triplet and a 5'-overhang that flips out and away from the helical axes to form a Hoogsteen base-pair with the 3'-overhang of a symmetry mate. The global conformations of the three structures differ, however, in the widths of their respective major grooves, the lengths of the molecules, and the extent of crystal packing. The structures were determined from crystals grown in an unusual precipitant for A DNA, polyethylene glycol (PEG) 400, in combination with polyamines or ions; cobalt hexamine for the pseudo-tridecamer, and spermidine for the pseudo dodecamers. As the Sp1 binding site is a target for antiviral and anticancer drugs, pseudo-dodecamer crystals were soaked with one such antiviral and anticancer compound, P4N. Although P4N was not visualized unambiguously in the electron density maps, the effect of the drug is evident from significant differences in the lattice constants, crystal packing, and overall conformation of the structure. PMID- 15896804 TI - Structural analysis of Escherichia coli ThiF. AB - Escherichia coli ThiF is an enzyme in the biosynthetic cascade for generating the essential cofactor thiamin pyrophosphate. In this cascade, ThiF catalyzes adenylation of the C terminus of ThiS. We report here the crystal structures of ThiF, alone and in complex with ATP. The structures provide insight into a preference for ATP during adenylation of the protein ThiS. Additionally, the structures reveal an ordered crossover loop predicted to clamp the flexible tail of ThiS into the ThiF active site during the adenylation reaction. The importance of the crossover loop for ThiF activity is highlighted by mutational analysis. Comparison of ThiF with the structural homologues MoeB, APPBP1-UBA3, and SAE1 SAE2 reveals that the ATP-binding site, including an arginine-finger, is maintained throughout evolution, and shows divergence occurring in protein substrate-binding sites and regions devoted to unique steps in the specific function of each enzyme. PMID- 15896805 TI - Atomic resolution structures of R-specific alcohol dehydrogenase from Lactobacillus brevis provide the structural bases of its substrate and cosubstrate specificity. AB - The R-specific alcohol dehydrogenase (RADH) from Lactobacillus brevis is an NADP dependent, homotetrameric member of the extended enzyme family of short-chain dehydrogenases/reductases (SDR) with a high biotechnological application potential. Its preferred in vitro substrates are prochiral ketones like acetophenone with almost invariably a small methyl group as one substituent and a bulky (often aromatic) moiety as the other. On the basis of an atomic-resolution structure of wild-type RADH in complex with NADP and acetophenone, we designed the mutant RADH-G37D, which should possess an improved cosubstrate specificity profile for biotechnological purposes, namely, a preference for NAD rather than NADP. Comparative kinetic measurements with wild-type and mutant RADH showed that this aim was achieved. To characterize the successful mutant structurally, we determined several, partly atomic-resolution, crystal structures of RADH-G37D both as an apo-enzyme and as ternary complex with NAD or NADH and phenylethanol. The increased affinity of RADH-G37D for NAD(H) depends on an interaction between the adenosine ribose moiety of NAD and the inserted aspartate side-chain. A structural comparison between RADH-G37D as apo-enzyme and as a part of a ternary complex revealed significant rearrangements of Ser141, Glu144, Tyr189 and Met205 in the vicinity of the active site. This plasticity contributes to generate a small hydrophobic pocket for the methyl group typical for RADH substrates, and a hydrophobic coat for the second, more variable and often aromatic, substituent. Around Ser141 we even found alternative conformations in the backbone. A structural adaptability in this region, which we describe here for the first time for an SDR enzyme, is probably functionally important, because it concerns Ser142, a member of the highly conserved catalytic tetrad typical for SDR enzymes. Moreover, it affects an extended proton relay system that has been identified recently as a critical element for the catalytic mechanism in SDR enzymes. PMID- 15896806 TI - The complement of enzymatic sets in different species. AB - We present here a comprehensive analysis of the complement of enzymes in a large variety of species. As enzymes are a relatively conserved group there are several classification systems available that are common to all species and link a protein sequence to an enzymatic function. Enzymes are therefore an ideal functional group to study the relationship between sequence expansion, functional divergence and phenotypic changes. By using information retrieved from the well annotated SWISS-PROT database together with sequence information from a variety of fully sequenced genomes and information from the EC functional scheme we have aimed here to estimate the fraction of enzymes in genomes, to determine the extent of their functional redundancy in different domains of life and to identify functional innovations and lineage specific expansions in the metazoa lineage. We found that prokaryote and eukaryote species differ both in the fraction of enzymes in their genomes and in the pattern of expansion of their enzymatic sets. We observe an increase in functional redundancy accompanying an increase in species complexity. A quantitative assessment was performed in order to determine the degree of functional redundancy in different species. Finally, we report a massive expansion in the number of mammalian enzymes involved in signalling and degradation. PMID- 15896807 TI - Vitamin B12, demyelination, remyelination and repair in multiple sclerosis. AB - Multiple Sclerosis (MS) and vitamin B12 deficiency share common inflammatory and neurodegenerative pathophysiological characteristics. Due to similarities in the clinical presentations and MRI findings, the differential diagnosis between vitamin B12 deficiency and MS may be difficult. Additionally, low or decreased levels of vitamin B12 have been demonstrated in MS patients. Moreover, recent studies suggest that vitamin B12, in addition to its known role as a co-factor in myelin formation, has important immunomodulatory and neurotrophic effects. These observations raise the questions of possible causal relationship between the two disorders, and suggest further studies of the need to close monitoring of vitamin B12 levels as well as the potential requirement for supplementation of vitamin B12 alone or in combination with the immunotherapies for MS patients. PMID- 15896808 TI - The therapeutic use of stem cells for myelin repair in autoimmune demyelinating disorders. AB - Spontaneous remyelination occurs in multiple sclerosis (MS) patients. However, this process is not robust enough to promote a functional and stable recovery of the myelin architecture in demyelinated areas of the central nervous system (CNS). As a consequence of this incomplete reparative process, the disease invariably progresses and patchy areas of demyelination-in which axonal damage and/or loss is a constant accompanying factor-increase over time and lead to the accumulation of irreversible neurological deficits. Thus, the development of cell based therapies aimed to promote multifocal remyelination in MS represents one of the most challenging areas of investigation. Several cell-replacement strategies have been developed in the last few years. However, most of these therapeutic approaches-although consistently able to form new myelin sheaths around the transplantation site-are unrealistic owing to the multifocality of the demyelinating process and the inability to in vitro growth and differentiate large number of myelin-forming cells. Recently, promising cell-replacement therapies based on the use of stem cells have been proposed. However, before envisaging any potential human applications of such therapies we need to confront with some preliminary and still unsolved questions: (i) the ideal stem cell source for transplantation, (ii) the route of cell administration, (iii) the differentiation and persistence of stem cells into the targeted tissue and, last but not least, (iv) the functional and long-lasting integration of transplanted cells into the host tissue. PMID- 15896809 TI - Neurofilament phosphoforms: surrogate markers for axonal injury, degeneration and loss. AB - This review on the role of neurofilaments as surrogate markers for axonal degeneration in neurological diseases provides a brief background to protein synthesis, assembly, function and degeneration. Methodological techniques for quantification are described and a protein nomenclature is proposed. The relevance for recognising anti-neurofilament autoantibodies is noted. Pathological implications are discussed in view of immunocytochemical, cell culture and genetic findings. With reference to the present symposium on multiple sclerosis, the current literature on body fluid levels of neurofilaments in demyelinating disease is summarised. PMID- 15896810 TI - Oxidative stress, mitochondrial dysfunction and cellular stress response in Friedreich's ataxia. AB - There is significant evidence that the pathogenesis of several neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease, Friedreich's ataxia (FRDA), multiple sclerosis and amyotrophic lateral sclerosis, may involve the generation of reactive oxygen species (ROS) and/or reactive nitrogen species (RNS) associated with mitochondrial dysfunction. The mitochondrial genome may play an essential role in the pathogenesis of these diseases, and evidence for mitochondria being a site of damage in neurodegenerative disorders is based in part on observed decreases in the respiratory chain complex activities in Parkinson's, Alzheimer's, and Huntington's disease. Such defects in respiratory complex activities, possibly associated with oxidant/antioxidant imbalance, are thought to underlie defects in energy metabolism and induce cellular degeneration. The precise sequence of events in FRDA pathogenesis is uncertain. The impaired intramitochondrial metabolism with increased free iron levels and a defective mitochondrial respiratory chain, associated with increased free radical generation and oxidative damage, may be considered possible mechanisms that compromise cell viability. Recent evidence suggests that frataxin might detoxify ROS via activation of glutathione peroxidase and elevation of thiols, and in addition, that decreased expression of frataxin protein is associated with FRDA. Many approaches have been undertaken to understand FRDA, but the heterogeneity of the etiologic factors makes it difficult to define the clinically most important factor determining the onset and progression of the disease. However, increasing evidence indicates that factors such as oxidative stress and disturbed protein metabolism and their interaction in a vicious cycle are central to FRDA pathogenesis. Brains of FRDA patients undergo many changes, such as disruption of protein synthesis and degradation, classically associated with the heat shock response, which is one form of stress response. Heat shock proteins are proteins serving as molecular chaperones involved in the protection of cells from various forms of stress. In the central nervous system, heat shock protein (HSP) synthesis is induced not only after hyperthermia, but also following alterations in the intracellular redox environment. The major neurodegenerative diseases, Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Huntington's disease (HD) and FRDA are all associated with the presence of abnormal proteins. Among the various HSPs, HSP32, also known as heme oxygenase I (HO-1), has received considerable attention, as it has been recently demonstrated that HO-1 induction, by generating the vasoactive molecule carbon monoxide and the potent antioxidant bilirubin, could represent a protective system potentially active against brain oxidative injury. Given the broad cytoprotective properties of the heat shock response there is now strong interest in discovering and developing pharmacological agents capable of inducing the heat shock response. This may open up new perspectives in medicine, as molecules inducing this defense mechanism appear to be possible candidates for novel cytoprotective strategies. In particular, manipulation of endogenous cellular defense mechanisms, such as the heat shock response, through nutritional antioxidants, pharmacological compounds or gene transduction, may represent an innovative approach to therapeutic intervention in diseases causing tissue damage, such as neurodegeneration. PMID- 15896811 TI - Establishment and characterization of continuous hematopoietic progenitors derived pig normal mast cell lines. AB - Mast cells (MCs) are tissue resident, hematopoietic stem cells-derived elements, distributed throughout the body. They are the pivotal mediating cells of allergic reactions. In addition, in mice, MCs play a critical role in the defense against several pathogens, such as bacteria, parasites and viruses. Whereas the biology of rodent and human MCs has been extensively studied using in vitro derived populations, the role of MCs in pigs has not yet been evaluated, given the very low availability of pure porcine MCs populations. In the present report, we describe an original method to obtain continuous factor-dependent normal pig MCs (PMC) lines from fetal hematopoietic progenitors. These Stem Cell Factor (SCF) and Interleukin-3- (IL-3)-dependent PMC lines retain their capacity to growth after conventional freezing methods and exhibit most of the morphological and biochemical properties of normal, although immature, MCs, including metachromatic granules containing sulfated polysaccharides, the expression of c-kit and high affinity IgE receptors (FcepsilonRI), and the ability to store histamine that is released upon cross-linking of FcepsilonRI. In vitro derived PMC lines might thus be valuable tools to further investigate the reactivity of these elements towards several parasites frequently encountered in pig, such as, but not limited to, Ascaris suum, Trichinella spiralis or Trichuris suis, or towards antigens derived from these pathogens. PMID- 15896812 TI - Effects of hydrocarbon contamination on a free living marine nematode community: results from microcosm experiments. AB - Anthropogenic inputs of crude and refined petroleum hydrocarbons into the sea require knowledge of the effects of these contaminants on the receiving assemblages of organisms. A microcosm experiment was carried out to study the influence of diesel on a free living nematode community of a Tunisian lagoon. Sediments were contaminated by diesel that ranged in concentration from 0.5 to 20 mg diesel kg(-1) dry weight (dw), and effects were examined after 90 days. Gradual changes in community structure were revealed depending on the quantity of diesel administrated. In the medium (1 mg diesel kg(-1) and 5 mg diesel kg(-1) (dw)) and high (10 mg diesel kg(-1), 15 mg diesel kg(-1) and 20 mg kg(-1) (dw)) treated microcosms, most univariate measures, including diversity and species richness, decreased significantly with increasing level of diesel contamination whereas nematode assemblage from the low treated microcosm (0.5 mg diesel kg(-1) (dw)) remained unaffected. Results from multivariate analyses of the species abundance data demonstrated that responses of nematode species to the diesel treatments were varied: Chaetonema sp. was eliminated at all doses tested and seemed to be intolerant species to diesel contamination; Pomponema sp. and Oncholaimus campylocercoides were significantly affected at all diesel contamination levels but they were not eliminated, these species were categorized as "diesel-sensitive"; Hypodontolaimus colesi, Daptonema trabeculosum and Daptonema fallax which significantly increased respectively at 0.5, 1 and 5 mg diesel kg(-1) (dw) concentrations and appeared to be "opportunistic" species at these doses whereas Marylynnia stekhoveni which increased at all high doses (10, 15 and 20 mg diesel kg(-1) (dw)) seemed to be a "diesel-resistant" species. PMID- 15896813 TI - Concentration of polychlorinated biphenyls (PCBs) in beached resin pellets: variability among individual particles and regional differences. AB - Concentrations of polychlorinated biphenyls (PCBs) in beached resin pellets were examined to reveal variability between individual particles and differences among beaches. Fifty-five resin pellets from a beach in Tokyo were individually analyzed for PCBs, and showed concentrations ranging from <28 to 2,300 ng/g. This indicates that concentrations are highly variable between particles. Among several characters, discoloration (e.g., yellowing) had a positive relationship with PCB concentration: discolored pellets contained more PCBs than others on most of the beaches sampled. Given the color-selective ingestion of food by some organisms, this may be ecotoxicologically important. Measurements of samples from 47 beaches in Japan showed regional differences in PCB concentrations in resin pellets consistent with those in mussels. Sporadic high concentrations of PCBs were also found in pellets from remote islands, suggesting that resin pellets could be the dominant route of exposure to the contaminants at remote sites. The similarity of PCB concentrations between resin pellets and mussels suggests a potential use of resin pellets to monitor pollution in seawater. PMID- 15896814 TI - Roles of the polymerase and BRCT domains of Rev1 protein in translesion DNA synthesis in yeast in vivo. AB - Rev1p in yeast is essential for the translesion of abasic sites and 6-4 photoproducts. It plays a role as a translesion polymerase, but also supports translesion catalyzed by other polymerases. The protein has two domains, BRCT and Y-family polymerase. A point mutation in the BRCT domain is known to abolish the second function. In the present research, we have studied the effects of deletion of the BRCT domain and a point mutation at the two amino acids in the putative polymerase active center. We have introduced an abasic site, its tetrahydrofuran analog, and a 6-4 thymine-thymine photoproduct using the oligonucleotide transformation assay. Translesion efficiencies were estimated from the transforming activities of the oligonucleotides with a lesion, and the mutation spectra were analyzed by DNA sequencing of the transformants. Results showed that the lack of the BRCT domain reduced translesion efficiencies, but that substantial translesion synthesis took place. The mutation spectra of the lesions were not greatly affected. Therefore, the BRCT domain may be important, but dispensable for translesion synthesis. In contrast, the polymerase mutation, rev1AA, has only small effects on the translesion efficiencies, but the mutation spectra were greatly affected; the incorporation of dCMP opposite the lesions was specifically lost. This clearly shows that the polymerase domain is responsible for the dCMP incorporation. The effect of Poleta was also analyzed. From all the results DNA polymerases other than these two translesion polymerases, too, seem to initiate the translesion synthesis. PMID- 15896815 TI - Visual mental imagery during caloric vestibular stimulation. AB - We investigated high-resolution mental imagery and mental rotation, while the participants received caloric vestibular stimulation. High-resolution visual mental imagery tasks have been shown to activate early visual cortex, which is deactivated by vestibular input. Thus, we predicted that vestibular stimulation would disrupt high-resolution mental imagery; this prediction was confirmed. In addition, mental rotation tasks have been shown to activate posterior parietal cortex, which is also engaged in the processing of vestibular stimulation. As predicted, we also found that mental rotation is impaired during vestibular stimulation. In contrast, such stimulation did not affect performance of a low imagery control task. These data document previously unsuspected interactions between the vestibular system and the high-level visual system. PMID- 15896816 TI - Cognitive and psychosocial correlates of alexithymia following traumatic brain injury. AB - Changes in emotional and social behaviour are considered to be amongst the most common and debilitating consequences of traumatic brain injury (TBI). Little is known of the effects of TBI on alexithymia, which refers to impairment in aspects of understanding emotions. In the current study TBI patients (N=28) were compared with demographically matched healthy controls (N=31) on the Toronto Alexithymia Scale-20 (TAS-20), a measure that taps three distinct characteristics of the alexithymia concept; difficulty in identifying emotions, difficulty in describing emotions and externally oriented thinking. Patients and controls also completed measures of anxiety, depression, quality of life, and measures of fluency to assess executive function. Patients showed greater levels of alexithymia, in terms of difficulty identifying emotions and reduced introspection. Difficulty in identifying emotions was associated with poorer quality of life, even when depression and anxiety were controlled. Difficulty in identifying emotions was also uniquely associated with executive function deficits. Thus, although studies typically focus on aspects of cognitive change following head injury, these results lend support to Becerra et al.'s (Becerra, R., Amos, A., & Jongenelis, S. (2002). Organic alexithymia: a study of acquired emotional blindness. Brain Injury, 16, 633-645.) notion of an 'organic alexithymia', and suggest that more attention should be focused upon assessment of emotional change post-head injury. PMID- 15896817 TI - Molecular cloning and characterization of genes encoding two microsomal oleate desaturases (FAD2) from olive. AB - Two different cDNA sequences, designated OepFAD2-1 and OepFAD2-2, encoding two microsomal oleate desaturases (FAD2) have been isolated from olive (Olea europaea cv. Picual) using a PCR approach. Both deduced amino acid sequences showed the three histidine boxes characteristic of all membrane-bound desaturases, and possess a C-terminal endoplasmic reticulum retention signal. Phylogenetic analysis shows that OepFAD2-1 and OepFAD2-2 are grouped with other plant FAD2 sequences. Functional expression of the corresponding FAD2 cDNAs in yeast confirmed that they encode microsomal oleate desaturases. Genomic Southern blot analysis is consistent with the presence of at least two copies of each OepFAD2 gene in the olive genome. OepFAD2-1 transcript was strongly detected in very young seeds and in leaves, showing low levels in mesocarps, while the transcript of the OepFAD2-2 gene was moderately expressed in developing seeds, ripening mesocarp and leaves. These expression data suggest differential functions for the two olive microsomal oleate desaturase genes, with FAD2-1 possibly responsible for the desaturation of reserve lipids in the young seed, while FAD2-2 may be mainly involved in storage lipid desaturation in the mature seeds and the mesocarp. PMID- 15896818 TI - New approach to 3-oxo-4-aza-5alpha-androst-1-ene-17beta-(N-tert butylcarboxamide). AB - We describe the synthesis of 3-oxo-4-aza-5alpha-androst-1-ene-17beta-(N-tert butylcarboxamide) (finasteride) from 4-androstene-3,17-dione (AD) in seven steps in an overall yield of 18.6% via oxidation, ammoniumation, dehydration, and dehydrogenation. PMID- 15896819 TI - Expression of novel organic cation/carnitine transporter (OCTN2) in the mouse pancreas. AB - Among the organic cation transporters, OCTN2 is identified as the most important carnitine transporter owing to the ability to transport carnitine. Although the OCTN2 is previously found in various tissues, there have been no reports showing the OCTN2 in the pancreas. In this study, we examined the expression and localization of OCTN2 in the mouse pancreas by the aid of an in situ hybridization technique and immunohistochemistry with anti-OCTN2 antibody. As a result, the OCTN2 expression was found in the A-cells for the first time. OCTN2 was not expressed in B-cells, notwithstanding that the metabolism of long-chain fatty acids, which are transported into the mitochondria with the help of carnitine, was expected for fatty acid-stimulated insulin secretion. Thus, this study suggests the possibility of carnitine uptake in the pancreatic A-cells through OCTN2 and implies the presence of carnitine transporter(s) other than OCTN2 in the B-cell. PMID- 15896820 TI - Circles are different: the perception of Glass patterns modulates early event related potentials. AB - Glass patterns are randomized dot arrays that generate the perception of a global structure. They consist of correlated dot pairs which are generated by geometric transformations. The present study employed behavioral and event-related brain potential (ERP) measures to characterize the underlying neuronal processing when such patterns are perceived. Stimuli were circular, parallel, and randomized Glass patterns presented in two isoluminant colors using a choice reaction paradigm. Sixteen subjects were instructed to differentiate between colors with a button-press response. The N170 component increased in amplitude for circular patterns, and this effect was most pronounced bilaterally over occipito-temporal areas. The results suggest that the global perception of form generated by Glass patterns occurs at a stage of visual processing past area V1. PMID- 15896821 TI - Adsorption of As(V) and As(III) by nanocrystalline titanium dioxide. AB - This study evaluated the effectiveness of nanocrystalline titanium dioxide (TiO(2)) in removing arsenate [As(V)] and arsenite [As(III)] and in photocatalytic oxidation of As(III). Batch adsorption and oxidation experiments were conducted with TiO(2) suspensions prepared in a 0.04 M NaCl solution and in a challenge water containing the competing anions phosphate, silicate, and carbonate. The removal of As(V) and As(III) reached equilibrium within 4h and the adsorption kinetics were described by a pseudo-second-order equation. The TiO(2) was effective for As(V) removal at pH<8 and showed a maximum removal for As(III) at pH of about 7.5 in the challenge water. The adsorption capacity of the TiO(2) for As(V) and As(III) was much higher than fumed TiO(2) (Degussa P25) and granular ferric oxide. More than 0.5 mmol/g of As(V) and As(III) was adsorbed by the TiO(2) at an equilibrium arsenic concentration of 0.6mM. The presence of the competing anions had a moderate effect on the adsorption capacities of the TiO(2) for As(III) and As(V) in a neutral pH range. In the presence of sunlight and dissolved oxygen, As(III) (26.7 microM or 2mg/L) was completely converted to As(V) in a 0.2g/L TiO(2) suspension through photocatalytic oxidation within 25 min. The nanocrystalline TiO(2) is an effective adsorbent for As(V) and As(III) and an efficient photocatalyst. PMID- 15896822 TI - Vertical fluxes and accumulation of PCBs in coastal sediments of the Rio de la Plata estuary, Argentina. AB - Settling particles and underlying sediments collected at 1, 2.5 and 4 km along offshore transects in the urbanized sector of the Rio de la Plata were analyzed to evaluate the sources and accumulation of PCBs. Total PCB concentrations range from <0.1 to 100 ng g(-1) and include variability associated to North-South and offshore gradients reflecting the impact of coastal discharges. Highest concentrations were recorded in the industrialized Central area close to Buenos Aires (61+/-37 ng g(-1) at 1 km) relative to cleaner northern stations (3.6+/-2.2 ng g(-1)) and southward sites (37+/-2.8 ng g(-1)), affected by transport of particulate PCBs by coastal currents. Sediment traps deployed in the Central area revealed large depositional fluxes of total matter (361+/-124 gm(-2)day(-1)) and PCBs (26+/-19 microg m(-2)day(-1)) and high sedimentation rates (5.0+/-1.7 cm yr( 1)). Uniform PCB concentrations (66-89 ng g(-1)) down to 20 cm in sediment cores suggest continued PCB discharges during the last 4 years. PCB composition was dominated by hexa (43+/-6.4%), hepta (23+/-5.1%) and pentachlorobiphenyls (21+/ 5.5%) with lower proportions tri-tetra (7.4+/-5.4%) and higher chlorinated congeners (5.1+/-3.3%). A consistent weathering pattern with loss of 3-5 chlorobiphenyls and enrichment in higher chlorinated PCBs corresponding to a shift from a 1:1 to a 1:3 1254:1260 Aroclor mixture, was observed offshore. A principal component analysis performed with the relative contribution of PCB congener classes confirmed the offshore weathering pattern indicating that transformer oils containing Aroclor 1254-1260 are the most probable sources. Sediment inventories, sediment trap fluxes and Fugacity II calculations indicate an accumulation approximately 500-800 kg PCB in superficial sediments of this coastal environment. PMID- 15896823 TI - A Neandertal mandible from the Cova del Gegant (Sitges, Barcelona, Spain). AB - A human mandible from the site of Cova del Gegant is described here for the first time and compared with other Middle and Upper Pleistocene representatives of the genus Homo from Europe and Southwest Asia. The specimen was recovered from sediments which also yielded Mousterian stone tools and Pleistocene fauna. The preserved morphology of the mandible, particularly in the region of the mental foramen, clearly aligns it with the Neandertals, making the Cova del Gegant the only known site in Catalonia documenting diagnostic human skeletal remains in association with Middle Paleolithic stone tools. This represents an important new addition to the human fossil record from the Iberian Peninsula and joins the Banolas mandible in documenting the course of human evolution in the northern Mediterranean region of Spain. PMID- 15896824 TI - Dietary restriction in C. elegans: from rate-of-living effects to nutrient sensing pathways. AB - The nematode Caenorhabditis elegans has been subjected to dietary restriction (DR) by a number of means, with varying results in terms of fecundity and lifespan. Two possible mechanisms by which DR increases lifespan are reduction of metabolic rate and reduction of insulin/IGF-1 signalling. Experimental tests have not supported either possibility. However, interaction studies suggest that DR and insulin/IGF-1 signalling may act in parallel on common regulated processes. In this review, we discuss recent developments in C. elegans DR research, including new discoveries about the biology of nutrient uptake in the gut, and the importance of invasion by the bacterial food source as a determinant of lifespan. The evidence that the effect of DR on lifespan in C. elegans is mediated by the TOR pathway is discussed. We conclude that the effect of DR on lifespan is likely to involve multiple mechanisms, which may differ according to the DR regimen used and the organism under study. PMID- 15896825 TI - Size fractionation of metals in runoff from residential and highway storm sewers. AB - Stormwater sampling for particulate, colloidal, and dissolved metals was conducted for several storms at six outfalls in Monmouth County, NJ. Samples were initially sequentially filtered through 5 microm, 0.45 microm, and 10 kDa filters. Of the heavy metals, Cu and Zn were mostly either dissolved (<10 kDa) (20-100%) or in the particulate size fractions >5 microm (0->70%). Pb and Cr were associated exclusively with particles >5 microm in size. Fe, Al, and Si were found mostly in larger size fractions (>70%), with smaller amounts 0.45-5 microm in size. Preliminary data from a small set of samples passed through coarser filters suggested that metals may actually be largely associated with particles larger than 20 microm. Variable and sometimes large dissolved fractions of Cu and Zn can contribute to erratic metals removal by structural best management practices (e.g., wet ponds, detention basins). The size fractionation of stormwater constituents has implications for the design and performance of stormwater control structures and the aquatic toxicity risks posed by the metals. The results demonstrate the importance of obtaining particle size data when planning stormwater treatment. PMID- 15896826 TI - Relation between platelet microaggregates and ankle brachial index in patients with peripheral arterial disease. AB - INTRODUCTION: Peripheral arterial disease is one of the systemic atherosclerotic diseases, and patients with the disorder are classified in the high risk group of coronary artery disease. A lower ankle brachial index is a frequent finding in peripheral arterial disease. While platelet microaggregates are a significant predictor of adverse clinical outcome in coronary artery disease, the significance of platelet aggregability in peripheral arterial disease has not been elucidated. MATERIALS AND METHODS: Small platelet aggregates measured using laser-light scattering and ankle brachial index were determined in 42 patients with both coronary artery disease and peripheral arterial disease (peripheral group), 56 patients with only coronary artery disease (coronary group) and 32 patients without both (control group). RESULTS: The level of small platelet aggregates was increased significantly in the peripheral group (4.3 x 10(4) [range 2.2 x 10(4) to 7.4 x 10(4)]) compared with both the coronary (1.1 x 10(4) [range 0.3 x 10(4) to 5.0 x 10(4)]) and control groups (0.5 x 10(4) [range 0.1 x 10(4) to 0.9 x 10(4)]). There was a significant inverse correlation between log small platelet aggregates and ankle brachial index (n=130, r=-0.422, p<0.001). Multivariate logistic regression analysis revealed that a lower ankle brachial index (<0.90) was an independent determinant of increased levels of small platelet aggregates. CONCLUSIONS: Platelet aggregability was increased in patients with peripheral arterial disease with the degree of platelet aggregation being closely associated with ankle brachial index. It is possible that this change in platelet activity may be one mechanism to explain why a lower ankle brachial index is a predictor of poor prognosis in patients with peripheral arterial disease. PMID- 15896827 TI - Effect of valsartan on monocyte/endothelial cell activation markers and adiponectin in hypertensive patients with type 2 diabetes mellitus. AB - Angiotensin II receptor blockade has been shown to have a beneficial effect on the angiopathies of hypertension and hyperglycemia in patients with type 2 diabetes. However, the effect of angiotensin II receptor blockade on monocyte and endothelial cell adhesion markers in type 2 diabetes is poorly understood. We investigated the effects of valsartan on these markers in 53 hypertensive patients with and without type 2 diabetes mellitus. Levels of monocyte activation markers (soluble CD14: 2.1+/-0.9 vs. 3.3+/-1.4 microg/ml, p<0.01; monocyte chemotactic peptide: 392+/-94 vs. 489+/-114 pg/ml, p<0.05; and monocyte-derived microparticles: 264+/-98 vs. 511+/-128/microL, p<0.01) and endothelial cell activation markers (soluble E-selectin: 41+/-11 vs. 61+/-20 ng/ml, p<0.001; and soluble vascular cell adhesion molecule-1: 478+/-82 vs. 584+/-101 ng/ml, p<0.01) were significantly increased in hypertensive patients with type 2 diabetes compared to normotensive controls. In addition, the concentrations of adiponectin were significantly decreased in patients with type 2 diabetes (8.1+/-3.1 vs. 5.2+/-2.5 microg/ml, p<0.01). Regardless of the presence of diabetic complications, both systolic and diastolic blood pressures significantly decreased after valsartan administration (valsartan 80 mg/day for 8 weeks). Monocyte and endothelial cell activation markers were decreased significantly in patients with type 2 diabetes after valsartan treatment, but not in non-type 2 diabetic patients. In addition, valsartan alleviated hypoadiponectinemia in hypertensive patients with diabetes (before vs. after: 5.2+/-2.5 vs. 7.6+/-2.7 microg/ml, p<0.001) but did not increase adiponectin levels in the non-diabetic hypertensive group, for which the average adiponectin level was normal prior to treatment. These results suggest angiotensin II receptor blockade (valsartan) may be beneficial as an anti-atherosclerotic therapy in patients with type 2 diabetes in addition to its anti-hypertensive action. PMID- 15896828 TI - Recurrent bilateral mammary fibromatosis (desmoid tumor) imaged with technetium 99m pentavalent dimercaptosuccinic acid [99mTc-(V)DMSA] scintimammography. AB - BACKGROUND: Breast fibromatosis is a rare, benign, recurring, locally destructive entity. CASE: A 35-year-old woman underwent right-sided lumpectomy, revealing fibromatosis with epithelial hyperplasia. Two years later, she was re-evaluated due to a three times as large mass recurrence on X-ray mammography. (99m)Tc (V)DMSA scintimammography revealed diffuse increased radiotracer uptake occupying the entire breast. A less widespread similar uptake was observed in the contralateral breast. Mammography did identify neither the posterior margins of the mass nor the left-sided lesion. The patient underwent mastectomy and left sided open biopsy, revealing bilateral fibromatosis. Ki-67 expression was moderate. CONCLUSIONS: (99m)Tc-(V)DMSA scintimammography can provide information regarding the nature of the lesion, its extent and bilateral involvement (especially in dense breasts) and could prove useful as a predictor of its potential for recurrence. PMID- 15896829 TI - Synchronous invasive squamous cell carcinoma and clear cell adenocarcinoma of the uterine cervix: a different human papillomavirus status. AB - BACKGROUND: A multiple primary invasive carcinoma of the cervix is a rare condition and is seldom composed of squamous cell carcinoma and clear cell adenocarcinoma. CASE: A 47-year-old woman presented with contact bleeding. The anterior lip of the cervix revealed a 2.0-cm protruded mass. Preoperative pathological and imaging studies demonstrated the squamous cell carcinoma of the exocervix alone. Radical hysterectomy was performed on the diagnosis of stage 1B cervical cancer. Histological examination of the specimen manifested a coexisting invasive clear cell adenocarcinoma in the endocervix. Human papillomavirus (HPV) 18 was detected in the squamous cell carcinoma; however, no HPV was detected in the clear cell adenocarcinoma. CONCLUSION: This finding suggests that there was an obvious difference in association of HPV with the two neoplasms. PMID- 15896830 TI - Novel techniques to improve split-thickness skin graft viability during vulvo vaginal reconstruction. AB - BACKGROUND: Split-thickness skin grafts are often utilized for vulvo-vaginal reconstruction. Unfortunately, infection and sloughing may occur in up to 22% of patients with standard techniques especially at irradiated recipient sites. CASES: We report seven cases of vulvo-vaginal reconstruction using split thickness skin grafts. In this series, we used fibrin tissue adhesives with and without vacuum-assisted closure devices to augment graft adherence and viability. We briefly describe the clinical history, surgical techniques, and outcomes of the cohort. CONCLUSION: Fibrin tissue adhesives and wound vacuum-assisted closure devices may improve the viability of split-thickness skin grafts during vulvo vaginal reconstruction. PMID- 15896831 TI - Long-term survival and disease recurrence in patients with primary squamous cell carcinoma of the vulva. AB - OBJECTIVES: To assess time to failure and sites of failure with extended follow up of patients with squamous cell carcinoma (SCC) of the vulva. METHODS: A retrospective analysis of 330 patients with primary SCC of the vulva treated at Mayo Clinic between 1955 and 1990 was conducted. The main outcome measures were the rates of treatment failure. The Kaplan-Meier method and the log-rank test were used to estimate the rates of overall survival, disease-free survival, and recurrence. The Cox proportional hazards model was used to assess independent variables as prognostic factors for treatment failure. RESULTS: All 330 patients in the cohort underwent lymphadenectomy; 113 patients (34.2%) had involvement of the inguinofemoral nodes and 88 patients (26.7%) had treatment failure. Treatment failures occurred more frequently in patients who presented with inguinal metastasis at the primary surgery and during the first 2 years of follow-up. After 2 years, both groups, with or without positive inguinal nodes, had similar treatment failure rates. Most patients with disease recurrence in the groin died within the first 2 years of follow-up. Involvement of the inguinal nodes was the main independent predictive factor for survival, disease recurrence, and metastasis. CONCLUSIONS: Most treatment failures occurred during the 2 years after initial surgical management. However, in 35% of patients, disease reoccurred 5 years or more after diagnosis, which demonstrates the need for long term follow-up. Complete ipsilateral or bilateral inguinofemoral lymph node dissection ensures a thorough evaluation and treatment of the groin. PMID- 15896832 TI - Metastasis from breast carcinoma to a tamoxifen-related endometrial polyp. AB - BACKGROUND: Metastasis of extragenital neoplasms to an endometrial polyp is rare and until now, only 6 cases of such involvement has been described. CASE: A 58 year-old woman, who had been diagnosed 4 years ago with infiltrating ductal breast carcinoma and treated with surgery and tamoxifen therapy, was admitted to the gynecology clinic because of endometrial thickening observed during a routine abdominal ultrasonographic examination. A total hysterectomy with bilateral salpingo-oophorectomy was performed. Pathological examination of the specimen showed a large polyp which microscopically showed clusters of cells with signet ring morphology within the polyp stroma. The positivity of tumor cells for GCDFP 15 supported the diagnosis of metastatic breast carcinoma to endometrial polyp. CONCLUSION: Metastatic breast carcinoma should be considered in the differential diagnosis of carcinomas with signet ring cell morphology involving uterus. PMID- 15896833 TI - Re: A need for laparoscopic evaluation of patients with endometrial carcinoma selected for conservative treatment by P. Morice et al. PMID- 15896834 TI - Fertility and recurrence results of conservative surgery for borderline ovarian tumors. AB - OBJECTIVES: To evaluate the fertility and recurrence outcomes in women treated with fertility-sparing surgery for borderline ovarian tumors. METHODS: A total of 142 patients with borderline ovarian tumors managed surgically from 1993 to 2004 were identified from gynecologic oncology and pathology files of SSK Ankara Maternity and Women's Health Teaching Hospital. Sixty-two of those patients who had conservative surgery were eligible for the study. Information was acquired by retrospective medical record review and patient interview. RESULTS: The observed recurrence rates after radical and fertility-sparing surgery were 0.0% and 6.5%, respectively. Four patients from the conservative surgery group developed recurrence, in contrast to none of the patients from the non-conservative surgery group. No disease-related deaths occurred in any group. In the conservatively managed group, ten women had successful pregnancies, with a total of 10 live births and 3 abortions. The mean duration of follow-up for the conservative surgery group was 44.3 months (range, 3-128). CONCLUSION: Fertility-sparing surgery for borderline ovarian tumors should be considered for women in the reproductive age group who desire preservation of fertility. Recurrence is noted significantly more often after this type of treatment and close follow-up is needed to detect recurrent disease. PMID- 15896835 TI - Stage-based expert systems to guide a population of primary care patients to quit smoking, eat healthier, prevent skin cancer, and receive regular mammograms. AB - BACKGROUND: Treating multiple health behavior risks on a population basis is one of the most promising approaches to enhancing health and reducing health care costs. Previous research demonstrated the efficacy of expert system interventions for three behaviors in a population of parents. The interventions provide individualized feedback that guides participants through the stages of change for each of their risk behaviors. This study extended that research to a more representative population of patients from primary care practice and to targeting of four rather than three behaviors. METHODS: Stage-based expert systems were applied to reduce smoking, improve diet, decrease sun exposure, and prevent relapse from regular mammography. A randomized clinical controlled trial recruited 69.2% of primary care patients (N = 5407) at home via telephone. Three intervention contacts were delivered for each risk factor at 0, 6, and 12 months. The primary outcome measures were the percentages of at-risk patients at baseline who progressed to the action or maintenance stages at 24-month follow-up for each of the risk behaviors. RESULTS: Significant treatment effects were found for each of the four behaviors, with 25.4% of intervention patients in action or maintenance for smoking, 28.8% for diet, and 23.4% for sun exposure. The treatment group had less relapse from regular mammography than the control group (6% vs. 10%). CONCLUSION: Proactive, home-based, and stage-matched expert systems can produce relatively high population impacts on multiple behavior risks for cancer and other chronic diseases. PMID- 15896836 TI - Speech perception and short-term memory deficits in persistent developmental speech disorder. AB - Children with developmental speech disorders may have additional deficits in speech perception and/or short-term memory. To determine whether these are only transient developmental delays that can accompany the disorder in childhood or persist as part of the speech disorder, adults with a persistent familial speech disorder were tested on speech perception and short-term memory. Nine adults with a persistent familial developmental speech disorder without language impairment were compared with 20 controls on tasks requiring the discrimination of fine acoustic cues for word identification and on measures of verbal and nonverbal short-term memory. Significant group differences were found in the slopes of the discrimination curves for first formant transitions for word identification with stop gaps of 40 and 20 ms with effect sizes of 1.60 and 1.56. Significant group differences also occurred on tests of nonverbal rhythm and tonal memory, and verbal short-term memory with effect sizes of 2.38, 1.56, and 1.73. No group differences occurred in the use of stop gap durations for word identification. Because frequency-based speech perception and short-term verbal and nonverbal memory deficits both persisted into adulthood in the speech-impaired adults, these deficits may be involved in the persistence of speech disorders without language impairment. PMID- 15896837 TI - Social and psychological characteristics of Greek secondary school students with learning difficulties. AB - In Greece there is an almost total lack of special education after the primary school (6th grade). This is a descriptive study that aimed to examine social, academic and psychological characteristics of secondary school students with a history of special placement during primary school. It compared 86 students with a placement (group A) with their 1284 schoolmates with no such placement history (group B) with respect to scores on School Adjustment Scale, Academic Motivation Self-Rating Inventory and Youth Self-Report. Group A presented more family social, academic and psychological problems such as: One-parent family, lower socioeconomic status, poor adjustment, low motivation and (especially for boys) high internalizing and externalizing scores. The high rates of failure and dropout of these students indicate that in Greece there is a need to set up appropriate services to help secondary school students with learning difficulties. PMID- 15896838 TI - Conformational properties of DNA containing (CCA)n and (TGG)n trinucleotide repeats. AB - We have used CD spectroscopy, polyacrylamide gel electrophoresis, and UV absorption spectroscopy to study conformational properties of DNA fragments containing (CCA)n and (TGG)n repeats, which are the most length-polymorphic microsatellite sequences of the human genome. The (CCA)n fragments are random single strands at neutral and alkaline pH but they fold into intramolecular intercalated cytosine tetraplexes at mildly acid pH values. More acid values stabilize intermolecular tetraplex formation. The behavior of (TGG)n repeats is more complex. They form hairpins or antiparallel homoduplexes in low salt solutions which, however, are transformed into parallel-stranded guanine tetraplexes at physiological KCl concentrations. Their molecularity depends on the repeat number: (TGG)4 associates into an octameric complex, (TGG)8 forms tetramolecular complexes. (TGG)n with odd repeat numbers (5, 7, and 9) generate bimolecular and tetramolecular tetraplexes. The only (TGG)7 folds into an intramolecular tetraplex at low KCl concentrations, which is antiparallel stranded. Moreover, the (TGG)(n) fragments provide various mutually slipped conformers whose population increases with salt concentration and with the increasing repeat number. However, the self-structures of both strands disappear in the presence of the complementary strand because both (TGG)n and (CCA)n prefer to associate into the classical heteroduplex. We suppose that the extreme conformational variability of the DNA strands stands behind the length polymorphism which the (CCA)n/(TGG)n repeats exhibit in the human genome. PMID- 15896839 TI - Microscopic structure of gelatin coacervates. AB - Microscopic structure of simple coacervates of gelatin having concentration approximately 130 g/l were studied at 25 degrees C by atomic force microscopy (AFM), rheology, small angle neutron scattering (SANS), UV absorption and circular dichroism (CD) techniques. The behavior of viscoelastic exponents Delta' and Delta'' of storage and loss modulii (G'(omega) approximately omega Delta', G''(omega) approximately omega Delta") revealed that, Delta' = 0.25+/-0.01 and Delta'' = 0.78+/-0.1 for coacervates. The mass fractal dimension 'd(f)' for coacervate was found to be 2.27, which attributed a compact heterogeneous network structure to the coacervates. This is supported by AFM pictures. The CD and UV absorption data indicated presence of helical structures inside the coacervates phase. SANS results showed the existence of a single length scale associated with this system identified as gelatin persistence length, zeta = 27+/-2 A. These studies indicate that the coacervate phase is a low dimensional dense heterogeneous material comprised of strongly interconnected triple helices which imparts a large storage modulus to this material. PMID- 15896840 TI - Favorable chitosan/cellulose film combinations for copper removal from aqueous solutions. AB - Three well-formed film combinations of chitosan, in the beta form and cellulose acetate biopolymer, having different proportions, have been synthesized and characterized by X-ray diffraction and infrared spectroscopy. The film having a 1.0/0.50 proportion presented 6.87 mmol of nitrogen atoms per gram of synthesized hybrid, with the highest affinity for adsorbing copper from aqueous solutions at 298+/-1K. The isotherm obtained in this adsorption showed a saturation plateau that corresponds to 1.92 mmol of copper per gram of hybrid. The energetic effects caused by copper ion interaction were determined through calorimetric titration at the solid-liquid interface in aqueous solution and gave a net thermal effect that enabled the calculation of the exothermic enthalpic values and the equilibrium constant. The complete thermodynamic data showed that the system is favored by exothermic enthalpies, negative Gibbs free energies and positive entropies. PMID- 15896841 TI - Establishment of a novel human B-CLL-like xenograft model in nude mouse. AB - We have developed a novel murine model for B-CLL by engrafting human prolymphocytic leukemia (PLL) or B-CLL cell line cells (JVM-3 and MEC-2 cell lines, respectively) into nude mice. Not only treatment of the mice was a prerequisite for the success of the graft, but also for the first time, females appeared to accept the cells more easily than males. Surprisingly, tumoral murine models for B-CLL could be established with PLL cells but not with B-CLL cells. JVM-3 cells were efficiently transplanted into nude mice through subcutaneous or intravenous routes. Irradiated female mice appeared to be the optimal recipients for tumor growth. Such murine models for human B-CLL may help the development of therapeutic agents. PMID- 15896842 TI - Molecular cloning and functional characterizations of chicken TL1A. AB - TL1A cDNA in chickens (ChTL1A), a homologue of human TL1A that belongs to the tumor necrosis factor (TNF) ligand super family, was cloned and characterized for its biological properties. The ChTL1A cDNA consisted of 1461 bp, coding for 239 amino acid residues and revealed a TNF ligand superfamily signature ([LV]-x [LIVM]-x3-G-[LIVMF]-Y-[LIVMFY]2-x2-[QEKHL]). This cDNA sequence also had a transmembrane domain between residues 34 and 53 (VLLCLLAVLLLALPIAYLLA). mRNA for ChTL1A was detected in most of the tissues and organs sampled and the expression was increased in spleen and abdominal adipose tissue following LPS injection. Recombinant chicken TL1A protein showed cytotoxic activity to the L929 cell line and cultured chicken fibroblast cells. Injection of the TL1A protein to broiler chicks resulted in a decrease in feed intake, an increase in nitric oxide production and rectal temperature. As an homologous sequence of mammalian TNF alpha has not been identified in the chicken genome database so far, the present data suggest that chicken TL1A possibly functions as a substitute for TNF-alpha. PMID- 15896843 TI - Characterization and expression analysis of bactericidal permeability-increasing protein (BPI) antimicrobial peptide gene from channel catfish Ictalurus punctatus. AB - Antimicrobial peptides are important components of host defenses against microbial invasions. Bactericidal permeability-increasing protein (BPI) is an antimicrobial peptide belonging to the lipid transfer/LPS-binding protein family. It serves important roles in defending against Gram-negative bacteria in the innate immune system. Here we report cloning of complete BPI cDNA from channel catfish (Ictalurus punctatus) by 5' RACE after obtaining the partial BPI cDNA sequence from EST analysis. The channel catfish BPI cDNA is 1640 bp in length with a 1428-bp open reading frame that encodes a protein of 475 amino acids. Catfish BPI gene shows high similarity with the BPI/LBP gene isolated from other teleost fish. As part of ongoing efforts in comparative genome analysis, we have assigned the catfish BPI gene to bacterial artificial chromosome (BAC) clones. Southern blot analysis on multiple BPI BAC clones indicated the presence of a single copy of the BPI gene in the catfish genome. Reverse transcription polymerase chain reaction (RT-PCR) analysis on healthy tissues showed that BPI was expressed in a wide range of tissues including head kidney, gill, skin, trunk kidney, brain, intestine, liver, muscle, ovary, spleen and stomach. The BPI gene was not developmentally expressed until 48 h after fertilization. Quantitative real time PCR (QRT-PCR) analysis indicated that the BPI gene expression was induced after challenge with Edwardsiella ictaluri, the causative agent of enteric septicemia of catfish (ESC). BPI upregulation peaked 3 days after challenge, mirroring the expression pattern of inflammatory chemokines in catfish, suggesting that it plays a role in the innate defense response. PMID- 15896844 TI - Effects of dexamethasone on peripheral blood mononuclear cell phenotype in weanling piglets. AB - The aim of this study was to evaluate the effect of dexamethasone treatment on the immune system of weanling piglets. Piglets were administered dexamethasone (DEX; 1mg/kg, IM) every 12h for 2 consecutive days (short-term experiment) or DEX (1mg/kg, IM) daily for 2 weeks (long-term experiment). The relative percentage of CD8(+) T cells in peripheral blood mononuclear cells (PBMCs) was significantly decreased (P<0.05) in both short- and long-term DEX-treated groups compared to their control groups. The percentage of IgM(+) cells in PBMCs of the long-term DEX-treated group was greatly increased (P<0.05) in comparison to the control group. The results of this study indicate that short-term DEX-treatment increases leucocyte function; however, long-term DEX-treatment depresses leucocyte function, especially that of CD8(+) T cells. PMID- 15896845 TI - Leptospirosis and pancytopenia: two case reports and review of the literature. AB - Besides the classical manifestations, leptospirosis may rarely occur with erythroid hypoplasia and/or pancytopenia. In this study, we reported two cases of leptospirosis presented with pancytopenia as the prevailing manifestation. In addition, the presence of pancytopenia in leptospirosis is reviewed. In both patients, the outcome was favourable as the bone marrow aplasia reversed completely after treatment with intravenous penicillin. In conclusion, this case study suggests that Leptospira infection should be included in the differential diagnosis of febrile pancytopenia, even in the absence of classical signs of severe disease as jaundice, meningitis, renal failure and pulmonary infiltrates. PMID- 15896846 TI - Pseudomonal sepsis with subcutaneous nodules in an HIV-infected individual with high CD4 counts. PMID- 15896847 TI - Pathophysiological basis for the prophylaxis of preeclampsia through early supplementation with antioxidant vitamins. AB - Preeclampsia (PE) is a multisystem disorder that remains a major cause of maternal and foetal morbidity and death. To date, no treatment has been found that prevents the development of the disease. Endothelial dysfunction is considered to underlie its clinical manifestations, such as maternal hypertension, proteinuria, and edema; however, the precise biochemical pathways involved remain unclear. A current hypothesis invokes the occurrence of oxidative stress as pathogenically important, as suggested by the fact that in PE, the placental and circulating levels of lipid peroxidation products (F2-isoprostanes and malondialdehyde [MDA]) are increased and endothelial cells are activated. A potential mechanism for endothelial dysfunction may occur via nuclear transcription factor kappa B (NF-kappaB) activation by oxidative stress. Alternatively, the idea that the antiangiogenic placental soluble fms-like tyrosine kinase 1 factor (sFlt1) is involved in the pathogenesis of this disease is just emerging; however, other pathophysiological events seem to precede its increased production. This review is focused on evidence providing a pathophysiological basis for the beneficial effect of early antioxidant therapy in the prevention of PE, mainly supported by the biological effects of vitamins C and E. PMID- 15896848 TI - Nuclear phospholipase C: involvement in signal transduction. AB - During the past years, several independent laboratories have highlighted the presence of nuclear signaling pathways based on lipid hydrolysis, which are not a mere duplication of those occurring at the plasma membrane. Among the enzymes of the cycle, nuclear phosphoinositide-specific phospholipase C (PI-PLC) has been analyzed quite extensively. In this context, PI-PLCbeta1 appears to play a key role as a check point in the G1 phase of the cell cycle. It has also been shown that its activation and/or up-regulation is upon the control of type 1 insulin like growth factor receptor (IGF-R) in both mouse fibroblast and myoblasts, suggesting that its signaling activity is essential for the normal behavior of the cell, at least in culture. The recent discovery of a possible involvement of the deletion of PI-PLCbeta1 gene in the progression of myelodysplastic syndrome (MDS) to acute myeloid leukemia (AML) in humans strengthens the contention that nuclear PI-PLC signaling is essential for physiological processes such as cell growth and differentiation. Even though PI-PLCbeta1 is present and does not translocate to eukaryotic nuclei, this organelle, even though only in some conditions contains also PI-PLCgamma1 which acts not only as a PI-PLC but also as guanine nucleotide exchange factor (GEF) for PI 3-kinase enhancer (PIKE) and is somehow linked to PI 3-kinase (PI3K) activity. Also members of PI-PLCdelta family are shuttling from the nucleus to the cytoplasm and return and are possibly involved in the control of cell growth. We must also take into account the presence in the nucleus of other phospholipases such as phospholipase A2 (PLA2) and phospholipase D (PLD), which also exert a signaling activity upon external stimuli. On the whole this review highlights the latest development in the PI-PLC cycle in the nucleus, which in terms of activation, regulation and down-stream targets differs substantially from that located at the plasma membrane. PMID- 15896849 TI - Estimation of free radical formation by beta-ray irradiation in rat liver. AB - In vivo free radical reactions in rat liver as a result of exposure to low-dose beta-radiation was evaluated with electron paramagnetic resonance (EPR) spectroscopy by monitoring the reduction of the nitroxyl spin probe after intravenous administration. The EPR signal intensity of a nitroxyl probe as a function of time in bile flow was monitored by cannulating the bile duct through the cavity of an X-band EPR spectrometer. The results show that the rate of nitroxyl signal loss was higher in rats whose livers were exposed to beta-rays compared to unexposed rats. However, the rate of signal loss was lower in animals whose organs were exposed to air by opening the abdominal cavity. In vitro experiments also showed that the nitroxyl EPR signal loss was greater in an atmosphere of nitrogen than in air. Results suggest that under low levels of tissue oxygen, exposure to beta-rays results in nitroxyl signal loss, which may be mediated by free radical dependent pathways. When tissue oxygen were higher, hydrogen peroxide mediated oxidation of hydroxylamine may predominate resulting in a signal loss of smaller magnitudes. This study shows possible evidence of reactive oxygen species formation by low-dose beta-ray irradiation in a living animal. PMID- 15896850 TI - Study on the interaction between DNA and protein induced by anticancer drug carboplatin. AB - The interaction of DNA and human serum albumin (HSA) in the presence of anticancer drug carboplatin was studied with piezoelectric quartz crystal impedance (PQCI) and electrochemistry techniques. In the PQCI analysis, the correlative parameters including the frequency (f0), the motional resistance (R(m)), and the static capacitance (C0) in the experiment were obtained and discussed in detail. Additionally, the kinetics parameters of the cross-linking process were investigated and a response kinetics model was deduced. The values of association rate constant k1, dissociation rate constant k(-1) and the reaction equilibrium constant K were estimated to be 1.895 mg/ml(-1) s(-1), 4.7 x 10(-5) s(-1) and 4.03 x 10(4) (mg/ml)-1, respectively. Furthermore, cyclic voltammetry (CV) and electrochemical AC impedance techniques were employed to testify the cross-linking process. PMID- 15896851 TI - The experimental toxicology of metallic mercury on the murine peripheral motor system: a novel method of assessing axon calibre spectra using the phrenic nerve. AB - The toxicology of metallic mercury on motor neurons and their processes requires further work to resolve controversial implications in the aetiology of human motor neuron disease (MND). The assessment of experimental neurotoxicity in the peripheral motor system is, however, technically problematic and difficult to interpret. The mean number of axons in a nerve can vary considerably due to a high degree of biological variation. Atrophy of large axons can appear as loss when, in fact, their numbers appear in smaller diameter axonal categories. We addressed these quantitative problems using the murine phrenic nerve (MPN), a mono-fascicular, predominantly motor nerve as a model system. One micrometer transverse sections of gluteraldehyde/osmium tetroxide fixed MPNs were stained for myelin using a silver technique. Axon areas were measured from digital images of the nerve in cross-section (ImagePro Plus software) and transformed to circular diameter equivalents, then displayed as frequency distributions. We found a high biological variation in the mean axon number between paired nerves within experimental groups. Therefore, axon diameter data within individuals group was pooled. Theoretical simulation of axonal degeneration, atrophy and hypertrophy of larger myelinated axons (also affected in MND) were modelled by manipulating the original data set. With this model, by comparing normal distributions, it is possible to distinguish axonal atrophy, degenerative loss, and hypertrophy as distinct pathological processes in the large calibre axon subgroup that are selectively vulnerable to metallic toxins such as mercury. PMID- 15896852 TI - SOX13 is up-regulated in the developing mouse neuroepithelium and identifies a sub-population of differentiating neurons. AB - In mammals, most of the twenty SOX (SRY HMG box) transcription factors are expressed during embryogenesis and play an important role in cell fate determination. We show here that SOX13 is expressed in the developing mouse brain and spinal cord from E12.5 to E15.5, where it is largely confined to the differentiating zone rather than to the proliferating zone. In particular, we found that SOX13 expression was activated in a subset of neural progenitors as they exit the cycle of mitosis, migrate away from the ventricular zone, and start to differentiate into neurons. The SOX13 protein always localized to the nuclei of the differentiating neuronal cells, consistent with a role for SOX13 as a transcription factor during neurogenesis. Our data suggest a role for SOX13 in the specification and/or differentiation of a specific subset of neurons in the developing central nervous system. PMID- 15896853 TI - Quantitative detection of Cucumber vein yellowing virus in susceptible and partially resistant plants using real-time PCR. AB - A method for the detection of Cucumber vein yellowing virus (CVYV) that combines reverse transcription with real-time PCR (SYBR((R)) Green chemistry) was developed using specific primers designed from a nucleotide sequence of the RNA polymerase gene (NIb) conserved among all the available CVYV strains. This method provided a linear assay over five to six orders of magnitude and reproducibly detected titres as low as 10(3) molecules of the target CVYV cDNA. Real-time PCR gave reproducible results for the quantification of CVYV in young leaves of susceptible and resistant cucumber landraces after mechanical inoculation. Significant differences in the starting amount of target cDNA were found between the analyzed genotypes, indicating differences in viral accumulation that correlated to their different levels of resistance. Real-time PCR results validated our previous findings using slot-blot hybridization, the dominance of the strong resistance to CVYV displayed by C.sat 10, and provided improved reliability and sensitivity of detection. This method has great potential in resistance breeding for germplasm screening, characterization of resistance mechanisms and genetic studies. PMID- 15896854 TI - Detection and quantification of hepatitis A virus in seawater via real-time RT PCR. AB - A real-time RT-PCR method utilizing SYBR Green chemistry was developed to detect and enumerate hepatitis A virus (HAV) in ocean water. Ocean water samples were taken at the Tijuana River mouth (Tijuana, Mexico) and Imperial Beach pier (1.4 km north of the Tijuana River mouth in San Diego, California) following four separate rain events. A total of eight samples were collected, one from each location, each consisting of 4 l of ocean water. Using conventional RT-PCR and primers based on the conserved sequence at the VP3-VP1 genes of HAV, a 247 bp cDNA was amplified from six out of eight rain event water samples. HAV cDNA (confirmed by sequence analysis) was cloned into a TOPO vector (Invitrogen, Carlsbad, CA), and four primer sets were designed for application in SYBR Green real-time RT-PCR. The water samples were shown to contain inhibitors that affected real-time RT-PCR amplifications, however diluting the cDNA solution enabled successful amplification. Using real-time RT-PCR, HAV could be detected in all eight samples. Depending on the rain event, the viral load in these samples varied from 90 to 3523 copies of HAV/L of ocean water near the mouth of the Tijuana River, and 347 to 2656 copies/l near the Imperial Beach pier. The sensitivity, quantitative ability and the high throughput nature of SYBR Green real-time RT-PCR will be useful in monitoring HAV contamination in seawater. PMID- 15896855 TI - Production and characterization of recombinant human anti-HBs Fab antibodies. AB - Recombinant human Fab antibodies were generated with different reactivities against the hepatitis B virus surface (HBs) antigen. To isolate the antibodies, a method was used that combined transformation of human B cells by Epstein-Barr virus (EBV) infection with a primer-vector system developed for isolating DNA fragments of human Ig Fab portions. With this method, monoclonal and oligoclonal cell lines producing anti-HBs antibodies were established and three anti-HBs Fab antibodies were isolated from two of these cell lines. From analysis of affinity characteristics, immunohistochemical activity, and cytolysis activity, these three Fab antibodies were classified into three different groups. The first group had high affinity for HBs, the second had the ability to kill HBV-infected cells, and the third was applicable to immunohistochemical staining with HBV-infected cells. The combined effect of these antibodies was also investigated by complement-dependent cytotoxicity assay. PMID- 15896856 TI - Effects of Zn pre-exposure on Cd and Zn bioaccumulation and metallothionein levels in two species of marine fish. AB - Zinc is an essential trace metal but also a potential toxicant to aquatic organisms. In this study, two juvenile marine fish species, the black sea bream Acanthopagrus schlegeli and the grunt Teraponjarbua, were pre-exposed to Zn either from waterborne (0.74-170 microg L-1) or dietary (39-5926 microg g-1) Zn for 1 or 3 weeks. The concentrations of Zn and metallothionein (MT) in the whole body of the black sea bream and in the gills, viscera and carcass of the grunt were then measured during this pre-exposure. Following the pre-exposure, both fish species were then exposed to 109Cd and 65Zn labeled food or water to quantify the dietary assimilation efficiency (AE) and the uptake rate of dissolved Cd and Zn. Zn concentrations in both fish species were enhanced after pre-exposure, but the increases were much less than the increase of ambient Zn pre-exposure concentration. Following Zn pre-exposure, MT concentrations in the viscera and carcass were significantly elevated, whereas the MT levels were not significantly elevated in the gills. Waterborne and dietary Zn exposure enhanced the uptake rates of dissolved Cd and Zn in both fish. The maximum increases of uptake rate constants of dissolved Cd and Zn were up to 1.9-2.8 and 2.1-2.6 times, respectively, in the seabream and grunt. In contrast, dietary assimilation efficiency of Cd and Zn was not significantly enhanced following Zn pre-exposure. A positive linear relationship was found between the uptake rate constants of dissolved metals and Zn or MT concentrations in the fish. The results suggested that Zn pre-exposure increased the potential of metal uptake from ambient water, but had little effect on dietary metal uptake. Furthermore, the Zn body concentration and metal uptake from the dissolved phase were significantly dependent on the fish body size. PMID- 15896857 TI - Lessons from tomographic studies of the mammalian Golgi. AB - Basic structure studies of the biosynthetic machinery of the cell by electron microscopy (EM) have underpinned much of our fundamental knowledge in the areas of molecular cell biology and membrane traffic. Driven by our collective desire to understand how changes in the complex and dynamic structure of this enigmatic organelle relate to its pivotal roles in the cell, the comparatively high resolution glimpses of the Golgi and other compartments of the secretory pathway offered to us through EM have helped to inspire the development and application of some of our most informative, complimentary (molecular, biochemical and genetic) approaches. Even so, no one has yet even come close to relating the basic molecular mechanisms of transport, through and from the Golgi, to its ultrastructure, to everybody's satisfaction. Over the past decade, EM tomography has afforded new insights into structure-function relationships of the Golgi and provoked a re-evaluation of older paradigms. By providing a set of tools for structurally dissecting cells at high-resolution in three-dimensions (3D), EM tomography has emerged as a method for studying molecular cell biology in situ. As we move rapidly toward the establishment of molecular atlases of organelles through advances in proteomics and genomics, tomographic studies of the Golgi offer the tantalizing possibility that one day, we will be able to map the spatio temporal coordinates of Golgi-related proteins and lipids accurately in the context of 4D cellular space. PMID- 15896858 TI - How do nonprofit hospitals manage earnings? AB - We hypothesize that, unlike for-profit firms, nonprofit hospitals have incentives to manage earnings to a range just above zero. We consider two ways managers can achieve this. They can adjust discretionary spending [Hoerger, T.J., 1991. 'Profit' variability in for-profit and not-for-profit hospitals. Journal of Health Economics 10, 259-289.] and/or they can adjust accounting accruals using the flexibility inherent in Generally Accepted Accounting Principles (GAAP). To test our hypothesis we use regressions as well as tests of the distribution of earnings by Burgstahler and Dichev [Burgstahler, D., Dichev, I., 1997. Earnings management to avoid earnings decreases and losses. Journal of Accounting and Economics 24, 99-126.] on a sample of 1,204 hospitals and 8,179 hospital-year observations. Our tests support the use of discretionary spending and accounting accrual management. Like Hoerger (1991), we find evidence that nonprofit hospitals adjust discretionary spending to manage earnings. However, we also find significant use of discretionary accruals (e.g., adjustments to the third-party allowance, and allowance for doubtful accounts) to meet earnings objectives. These findings have two important implications. First, the previous evidence by Hoerger that nonprofit hospitals show less variation in income may at least partly be explained by an accounting phenomenon. Second, our findings provide guidance to users of these financial statements in predicting the direction of likely bias in reported earnings. PMID- 15896859 TI - The human Retinal Functional Unit. AB - It has long been known that readers of this page will move their eyes from one fixation to the next two to four times per second. It follows from this fact that each fixation triggers a unique optic nerve volley lasting up to 300 ms that contains all the information the retina processes between fixations. Here we give such volleys a name (Retinal Functional Unit, RFU) and use human subjects and interstimulus interval (ISI) experiments to define some of their properties. We report that RFUs can be dissected into an initial fraction that reaches the cortex and a later fraction that may not, depending on the ISI between successive stimuli. During the dissection process the perceptions of the stimuli change in an orderly way, such that successive thresholds of "twoness", color, and duration are reached as a function of increasing ISI. We conclude that volleys from the tens or hundreds of thousands of active axons contained in every RFU exit the retina in a precisely determined temporal order, and add this conclusion to three others for which we have already published the supporting data. 1) The mammalian retina normally takes about 300 ms to process a visual stimulus. 2) The ca. 300 ms end product, an RFU, contains in neuronal form all the photochemical information acquired during one fixation. 3) These information-rich volleys reach the cortex with little or no change thanks to monosynaptic transfer in the thalamus. PMID- 15896860 TI - Event-related brain potentials and affective responses to threat in spider/snake phobic and non-phobic subjects. AB - We investigated cortical responses and valence/arousal ratings of spider phobic, snake phobic, and healthy subjects while they were processing feared, fear relevant, emotional neutral, and pleasant stimuli. Results revealed significantly larger amplitudes of late ERP components (P3 and late positive complex, LPC) but not of early components (N1, P2, N2) in phobics when subjects were processing feared stimuli. This fear-associated increase of amplitudes of late ERP components in phobic subjects was maximal at centro-parietal and occipital brain sites. Furthermore, phobics but not controls rated feared stimuli to be more negative and arousing than fear-relevant, emotional neutral, and pleasant stimuli. Since late ERP components and valence/arousal ratings were only significantly increased when phobic subjects but not when healthy controls were processing feared stimuli, the present data suggest that P3 and LPC amplitudes represent useful neural correlates of the emotional significance and meaning of stimuli. PMID- 15896861 TI - GPS analysis of human locomotion: further evidence for long-range correlations in stride-to-stride fluctuations of gait parameters. AB - During free walking, gait is automatically adjusted to provide optimal mechanical output and minimal energy expenditure; gait parameters, such as cadence, fluctuate from one stride to the next around average values. It was described that this fluctuation exhibited long-range correlations and fractal-like patterns. In addition, it was suggested that these long-range correlations disappeared if the participant followed the beep of metronome to regulate his or her pace. Until now, these fractal fluctuations were only observed for stride interval, because no technique existed to adequately analyze an extended time of free walking. The aim of the present study was to measure walking speed (WS), step frequency (SF) and step length (SL) with high accuracy (<1 cm) satellite positioning method (global positioning system or GPS) in order to detect long range correlations in the stride-to-stride fluctuations. Eight participants walked 30 min under free and constrained (metronome) conditions. Under free walking conditions, DFA (detrended fluctuation analysis) and surrogate data tests showed that the fluctuation of WS, SL and SF exhibited a fractal pattern (i.e., scaling exponent alpha: 0.5 < alpha < 1) in a large majority of participants (7/8). Under constrained conditions (metronome), SF fluctuations became significantly anti-correlated (alpha < 0.5) in all participants. However, the scaling exponent of SL and WS was not modified. We conclude that, when the walking pace is controlled by an auditory signal, the feedback loop between the planned movement (at supraspinal level) and the sensory inputs induces a continual shifting of SF around the mean (persistent anti-correlation), but with no effect on the fluctuation dynamics of the other parameters (SL, WS). PMID- 15896862 TI - Stability and variability may respond differently to changes in walking speed. AB - In gait research it has often been assumed that variability and stability are negatively correlated, where increases in variability are assumed to equate with increases in instability. The purpose of this paper is to illustrate that variability does not always equate with stability. To proof this point, a method was developed to directly assess stability and variability during the application of a visual perturbation at different walking speeds. Walking variability was measured by using the average standard deviation of the knee joint angle across the gait cycle. Walking stability was measured by the recovery time of the knee joint angle trajectory from the distortion induced by a visual perturbation that was delivered at the beginning of the stance phase. Five participants were required to walk at six different velocities on a treadmill (0.67, 0.80, 0.94, 1.07, 1.21, and 1.34 m/s). The coefficients of intraclass correlations for the experiment were 83% and 80% for the calculated stability and variability, respectively. The calculated stabilities were not sensitive to changes in walking speed (p>0.98). The calculated variability however decreased with increases in walking speed (p=0.004). No significant correlation between variability and stability was observed (r=-0.002). We suggest that gait stability is independent of variability during locomotion and should thus be measured independently. PMID- 15896863 TI - The locus of adaptation to altered gain in aimed movements. AB - Altered gain settings cause a mismatch between the actual movement amplitude across the surface and the distance covered on a real time visual display. The present study pursued three objectives of how adaptation to altered gain affects aimed motor behavior. First, we replicated findings of an earlier study reporting a negative linear relation between gain and both target acquisition time and end point variability. This means that our data do not agree with the classic U shaped relation between gain and acquisition time. Second, our results proved to be robust when we manipulated movement difficulty by varying target distance. And third, dividing a movement into four successive sections on the basis of key kinematic events revealed the locus of the adaptation to altered gain within movement execution. Time differences between gain conditions proved to start at a very early part of the movement, but appeared to be absent in the final movement section. In contrast, differences between gain conditions regarding the use of online feedback were also present in the last part of the movement during which the final target approach takes place. PMID- 15896864 TI - Coordination in arm movements during crawl stroke in elite swimmers with a loco motor disability. AB - The purpose of this study was to investigate selected kinematics parameters of the arm stroke in crawl swimmers with disabilities and to examine the potential use of an index of arm coordination (IdC) to evaluate the stroking technique of swimmers with diverse functional abilities. The degree of overlap in the propulsive phases (superposition model) and lag time between the propulsive phases (catch-up model) was examined in 18 well-trained swimmers with loco-motor disabilities, 9 females and 9 males, from functional classes S3-S10 with S10 being most functional. Based on the results, correct coordination appears to be fundamental to swimming crawl stroke in both able-bodied swimmers as well as swimmers with a disability. Some swimmers with disabilities examined here exhibited extreme values at both ends of the index scale. This might be essential to maintaining balance while swimming when not all limb activity contributes to the forward movement. PMID- 15896865 TI - Intravenous administration of human neural stem cells induces functional recovery in Huntington's disease rat model. AB - An animal model induced by striatal quinolinic acid (QA) injection shows ongoing striatal degeneration mimicking Huntington's disease. To study the migratory ability and the neuroprotective effect of human neural stem cells (NSCs) in this model, we transplanted NSCs (5 x 10(6)) or saline intravenously at 7 days after unilateral QA injection. NSCs-group exhibited the reduced apomorphine-induced rotation and the reduced striatal atrophy compared to the control. PCR analysis for the human-specific ERV-3 gene supported an evidence of the engraftment of human NSCs in the rat brain. X-gal+ cells were found in and around the damaged striatum and migrated NSCs differentiated into neurons and glias. This result indicates that intravenously injected human NSCs can migrate into the striatal lesion, decrease the following striatal atrophy, and induce long-term functional improvement in a glutamate toxicity-induced striatal degeneration model. PMID- 15896866 TI - Chronic stimulation of GABAA receptor with muscimol reduces amyloid beta protein (25-35)-induced neurotoxicity in cultured rat cortical cells. AB - The present study was performed to examine how the stimulation of gamma aminobutyric acid (GABA) receptor affects amyloid beta protein (25-35) (Abeta (25 35)), a synthetic 25-35 amyloid peptide, -induced neurotoxicity using cultured rat cortical neurons. Abeta (25-35) produced a concentration-dependent reduction of cell viability, which was significantly reduced by (5R,10S)-(+)-5-methyl-10,11 dihydro-5H-dibenzo[a,d] cyclohepten-5,10-imine (MK-801), an N-methyl-d-aspartate (NMDA) receptor antagonist, verapamil, an L-type Ca(2+) channel blocker, and N(G) nitro-l-arginine methyl ester (l-NAME), a nitric oxide synthase inhibitor. Pretreatment with muscimol, a GABAA receptor agonist, over a concentration range of 0.1-10microM 24h before the treatment with 10microM Abeta (25-35) showed concentration-dependent inhibition on the Abeta (25-35)-induced neuronal apoptotic death. However, baclofen (1 and 10microM), a GABAB receptor agonist, failed to inhibit the Abeta (25-35)-induced neuronal death. In addition, pretreatment with muscimol (1microM) for 24h inhibited the Abeta (25-35) (10microM)-induced elevation of cytosolic Ca(2+) concentration ([Ca(2+)]c) and glutamate release, generation of reactive oxygen species (ROS), and caspase-3 activity in cultured neurons. These neuroprotective effects of muscimol (1microM) were completely blocked by the simultaneous treatment with 10microM bicuculline, a GABAA receptor antagonist, indicating that the protective effects of muscimol were due to GABAA receptor stimulation. When, however, treated just 15min before the treatment with Abeta (25-35), muscimol (1microM) did not show any protective effect against Abeta (25-35) (10microM)-induced neurotoxicity in cultured neurons. These results suggest that the chronic activation of GABAA receptor may ameliorate Abeta-induced neurotoxicity by interfering with the increase of [Ca(2+)]c, and then by inhibiting glutamate release, generation of ROS and caspase-3 activity. PMID- 15896867 TI - Optimising two-dye microarray designs for estimating associations with a quantitative trait. AB - We discuss how the samples should be arranged in two-dye microarray studies when the objective is to investigate associations between gene expression and quantitative traits measured on each sample. Because there is typically large between array variation, information about the association will come from the differences in traits and expression measurements between the two values hybridised to the two dyes on the same array. It is shown that within-slide correlation of trait values should be minimised. The arrangement of samples for which this occurs will depend on the trait values in question, and is a computationally demanding problem. An alternative is to minimise the rank correlation. We discuss this and related issues for different combinations of numbers of samples and arrays. Data analysis, including estimation of the variance components, is also described. PMID- 15896868 TI - Evaluation of coupled convective-diffusive transport of drugs administered by intravitreal injection and controlled release implant. AB - A 3-dimensional finite element model was developed to simulate pharmacokinetics in the eye following drug administration by intravitreal injection and implant for the treatment of retinal disease. The contributions of (1) convection to the transport of drug through the vitreous and aqueous humor and (2) diffusion of drug in the vitreous were varied to study the drug elimination from a normal and diseased eye. Drug distribution achieved by intravitreal injection was compared to that for the same dose released at a constant rate over 15 h from an implant. The model was constructed for a rabbit eye and validated with experimental data for intravitreal injection of fluorescein. The implant reduced peak concentration by 43% and increased residence time by 71% for the baseline (6x10(-6) cm2/s drug diffusivity in vitreous and 0.1 microL/min vitreous outflow), when compared with that of intravitreal injection. Therefore, the implant could be beneficial in reducing the peak concentration and sustaining release of the drug for a longer duration. Convection has a relatively small influence in the normal eye for high diffusivity drugs (1x10(-5) cm2/s), but could have a significant effect for low diffusivity drugs (1x10(-7) cm2/s) in pathophysiologically elevated fluid outflow across the retina. By interpolating the results of this benchmark study, one could estimate the distributions for drugs of different molecular weight, and assess the effect of variable vitreous outflows associated with different pathophysiological conditions. PMID- 15896869 TI - Virus and transaminase levels determine the emergence of drug resistance during long-term lamivudine therapy in chronic hepatitis B. AB - BACKGROUND/AIMS: The results of earlier studies on determinants for the emergence of tyrosine-methionine-aspartate-aspartate (YMDD) mutants (rtM204 I/V) were controversial. The aim was to evaluate the impact of viral factors, host factors, host-viral interaction and drug factor on the emergence of rtM204 I/V. METHODS: 56 non-cirrhotic and 58 cirrhotic patients received lamivudine therapy for a median of 34 (12-60) months. RESULTS: rtM204 I/V emerged in 37 noncirrhotic and 36 cirrhotic patients. Stepwise logistic regression analysis showed that baseline hepatitis B e antigen (HBeAg) status [odds ratio (OR), 7.728; 95% confidental interval (CI), 2.886-12.957; P=0.0026], HBV-DNA level (OR, 3.756; 95% CI, 1.058 5.089; P=0.0202), alanine transaminase (ALT) level (OR, 6.285; 95% CI, 1.057 11.990; P=0.00246) and treatment duration (OR, 19.88; 95% CI, 8.652-31.762; P<0.0004) were independent determinants for the emergence of rtM204 I/V. Further categorical analysis and correlation test disclosed that patients with HBeAg positivity, HBV-DNA>500 pg/ml and ALT <5x upper limit of normal had significantly higher mutation rates. CONCLUSIONS: HBeAg status, HBV-DNA, ALT levels and treatment duration are the major determinants for the YMDD mutation during lamivudine therapy, and should be considered in designing the therapeutic strategy. PMID- 15896870 TI - Economic stability and health status: evidence from East Asia before and after the 1990s economic crisis. AB - The East Asian economies of Indonesia, Malaysia and Thailand suffered declines in their economic growth rates in 1997. The Indonesian and Thai government followed the World Bank prescription for adjustment, which included a cut-back in government spending at a time when there were significant job losses. Malaysia chose its own path to adjustment. Evidence presented in this paper shows that although the declines were short-lived that there was an impact on the health status measured by mortality rates for the populations of Indonesia and Thailand. There was little apparent impact on the health status of Malaysians. The lessons for other developing economies include the importance of social safety nets and the maintenance of government expenditure in minimising the impact of economic shocks on health. PMID- 15896871 TI - Myocardial border detection by branch-and-bound dynamic programming in magnetic resonance images. AB - Dynamic programming (DP) is a mathematical technique for making optimal decisions on the sequencing of interrelated problems. It has been used widely to detect borders in magnetic resonance images (MRI). MRI is noninvasive and generates clear images; however, it is impractical for manual measurement of the huge number of images generated by dynamic organs such as those of the cardiovascular system. A fast and effective algorithm is essential for on-line implementation of MRI-based computer aided measurement and diagnosis. In this paper, a branch-and bound dynamic programming technique is applied to detect the endocardial borders of the left ventricular. The proposed branch-and-bound method drastically reduces the computational time required in conventional exhaustive search methods. Statistical tests are conducted to verify the CPU time performance of the branch and-bound technique in comparison to the conventional exhaustive search method. PMID- 15896872 TI - Regulation of glutamate transporter GLAST and GLT-1 expression in astrocytes by estrogen. AB - Estrogen influences neuronal development and a broad spectrum of neural functions. In addition, several lines of evidence suggest a role as neuroprotective factor for estrogen in the CNS. Neuroprotection can result from direct estrogen-neuron interactions or be mediated indirectly involving the regulation of physiological properties of nonneuronal cells, such as astrocytes and microglia. Increased l-glutamate levels are associated with neurotoxic and neurodegenerative processes in the brain. Thus, the removal of l-glutamate from the extracellular space by astrocytes through the astroglial glutamate transporters GLT-1 and GLAST appears essential for maintaining a homeostatic milieu for neighboring neurons. We have therefore studied the influence of 17beta estradiol on l-glutamate metabolism in cultured astrocytes from the neonate mouse midbrain using quantitative RT-PCR and Western blotting for both transporters as well as functional l-glutamate uptake studies. The administration of estrogen significantly increased the expression of GLT-1 and GLAST on the mRNA and protein level. Likewise, specific l-glutamate uptake by astrocytes was elevated after estrogen exposure and mimicked by dbcAMP stimulation. Induction of transporter expression and l-glutamate uptake were sensitive to ICI 182,780 treatment suggesting estrogen action through nuclear estrogen receptors. These findings indicate that estrogen can prevent l-glutamate-related cell death by decreasing extracellular l-glutamate levels through an increased l-glutamate uptake capacity by astrocytes. PMID- 15896873 TI - Production of vaccines against leading biowarfare toxins can utilize DNA scientific technology. AB - There are a significant number of different natural toxins that are potential biological warfare agents against which a vaccine is needed. DNA science has been a key to the development of potential vaccines against the top threat toxin and should contribute such effects for other toxin's vaccines. Several different DNA technological scientific techniques have been used to accomplish the general goals of (1) cloning of the toxin or large toxin fragments, (2) altering the specific gene sequence to obtain high level expression of vaccine candidate production in alternate species (3) placement of the vaccine gene in very different presentation types of species. PMID- 15896874 TI - Eight contributions to social food studies in the low countries. PMID- 15896875 TI - Perceived naturalness and acceptance of genetically modified food. AB - This study examines people's acceptance of genetically modified (GM) food. Results suggest that GM acceptance depends most on how natural the genetically modified product is perceived and not directly on how natural the non-GM product is seen. A GM product that is perceived as more natural is more likely to be accepted than a GM product that is perceived as less natural. The extent to which GM affects the perceived naturalness of a product partly depends on the kind of product. PMID- 15896876 TI - Stereotypical thinking about foods and perceived capacity to promote weight gain. AB - The modern health media as well as the food and diet industries praise certain foods and food nutrients as being healthful while at the same time criticizing other foods and nutrients as promoters of obesity and disease. Do the categorical messages that much of the general public has assimilated concerning food influence judgements of the weight-enhancing properties of foods? In the present study a sample of adult participants (mostly middle-aged) rated the weight enhancing characteristics of a group of snack names that possess positive health reputations (e.g. a banana) along with snack names that were more disreputable in terms of wholesomeness (e.g. bacon). The results indicated that lower-calorie (and in some cases lower-fat) disreputable snacks were generally perceived to promote greater weight gain than much higher-calorie (and in some cases higher fat) reputable snacks. Beliefs about particular foods' goodness or badness as well as fat content were most often emphasized. The good versus bad message that Americans have assimilated concerning foods may be contributing to tendencies toward obesity. PMID- 15896877 TI - Food recommendations, tradition and change in a Flemish cookbook: Ons Kookboek, 1920-2000. AB - The first edition of Ons Kookboek (Our Cookbook), also known as 'the culinary bible of Flanders', was published in 1927 by the Belgian Women Farmers' Union. By 2000, an estimated 2.3 million copies had been sold. This exceptionally successful and long-running series of cookbooks represents a unique source of the history of Flemish cuisine and food culture. The aim of this article is to analyse how the food recommendations in Ons Kookboek changed during the past decades, parallel with the shrinking importance of the Belgian agricultural sector. Initially, the main goal of the editors was to overcome and to alter the monotonous nature of the rural menu through the giving of information about healthy, simple and inexpensive cooking, in which own produce occupied a central place. From the mid 1960s onwards, with the growing purchasing power and internationalisation of cooking, the content changed. Ons Kookboek was no longer only a basic cookbook with strictly didactic advice. The reader was given greater freedom and was encouraged to choose from a large culinary range, which resulted in contradictory recommendations. But notwithstanding all innovations, the link with the farming sector remained as the attitude towards a vegetarian or low-meat diet clearly demonstrates. PMID- 15896878 TI - Self-set dieting rules: adherence and prediction of weight loss success. AB - The low achievement rates among dieters could be partially due to ineffective dieting strategies, or an inability to adhere to them in the long term. The present research examines the effectiveness of self-set dieting rules for achieving dieting goals. Specifically, it investigates how the types of dieting rules and the extent to which dieters adhere to them predict dieting success. A prospective study with reassessment after two months was conducted and analyzed in 2003. Participants were 132 currently dieting females recruited on a university campus. They completed measures assessing self-set dieting rules, dieting goal, and height and weight at Time 1 and Time 2. Data were analyzed using chi(2) tests and multiple linear regression. Overall, adherence to self-set dieting rules was markedly low. Reduction of caloric intake and increase of exercise predicted weight loss success. Sustained adherence to the dieting rules 'reducing calories' and 'increasing exercise' was found to be effective for achieving self-set dieting goals. The findings suggest that encouraging long-term adherence to effective rules could help increase the number of successful self controlled dieting efforts. PMID- 15896879 TI - Disappearance of the traditional meal: temporal, social and spatial destructuration. AB - The disappearance of the 'traditional' meal was explored, comparing Flemish time budget data for 1988 and 1999. In 1988, 463 respondents between 21 and 40 years old kept a diary for three consecutive days. In 1999, 599 respondents followed the same procedure for a full week. Respondents registered all their activities, including timing, duration, location and other parties to the interaction. Respondents also completed a questionnaire. Questionnaire and time budget data from the 1988 and 1999 surveys were merged into a single, combined database. Separate analyses were undertaken for weekdays, Saturdays and Sundays. Data from 1988 and 1999 were compared, bearing in mind the temporal, social and spatial features of the meal. The results indicate that the Flemish are not eating indiscriminately, in terms of time, place or the company of others. There was no sign of a shift between 1988 and 1999 towards disorganisation of the meal in these terms. All in all, Flemish eating practices show a high level of structure in the temporal, spatial and social organisation of the meal. PMID- 15896880 TI - Nanocrystalline silver dressings as an efficient anti-MRSA barrier: a new solution to an increasing problem. AB - The emergence of multi-drug-resistant strains of bacteria represents a particular challenge in the field of wound management. The aim of the current study was to investigate whether nanocrystalline silver dressings possess the physical properties to act as a barrier to the transmission of methicillin-resistant Staphylococcus aureus (MRSA) in the laboratory setting and in a clinical setting. Initially, MRSA suspension and colony culture experiments were performed showing that nanocrystalline silver dressings act as potent and sustained antimicrobial agents, efficiently inhibiting MRSA penetration. Subsequently, a double-centre clinical trial was initiated using nanocrystalline silver dressings as a cover for 10 MRSA colonized wounds in a total of seven patients. By delineating the MRSA load on the upper side of the dressing and the wound bed each time the dressing was changed (i.e. after 1, 24, 48 and 72 h), nanocrystalline silver dressings were found to provide a complete, or almost complete, barrier to the penetration/spread of MRSA in 95% of readings. In addition, 67% of all wound observations showed a decrease in the MRSA load with an eradication rate of 11%. We believe that nanocrystalline silver dressings may become an important part of local MRSA management, with cost benefits to both patients and the healthcare system. PMID- 15896881 TI - Decreased agonist, but not antagonist, binding to the naturally occurring Thr92Lys variant of the h5-HT7(a) receptor. AB - In the present study on transfected human embryonic kidney (HEK)293 cells, we aimed at establishing whether expression of the naturally occurring Thr92Lys variation of the Gs-coupled h5-HT7(a) receptor leads to changes of ligand binding properties, of agonist-evoked cAMP formation and/or of antagonist-mediated blockade of the latter. Binding of [3H]5-carboxamidotryptamine ([3H]5-CT) to membranes and stimulated [3H]cAMP accumulation in whole cells were determined. Saturation binding experiments in membranes of transiently transfected cells expressing either the wild-type or the variant receptor revealed a single binding site in both cases and no difference in Bmax between both receptor isoforms. In competition binding experiments in membranes of stably transfected cells, the Thr92Lys variant exhibited a 2.8-11 times lower binding affinity of the ligands 5 hydroxytryptamine (5-HT), 5-CT, 5-methoxy-3-(1,2,3,6-tetrahydropyridin-4yl)-1H indole (RU24969), (+/-)-hydroxy-2-(di-n-propylamino)tetralin hydrobromide (8-OH DPAT) and sumatriptan compared to the wild-type receptor. However, the variant did not differ from the wild-type with respect to the binding properties of the antagonists (R)-3-(2-(2-(4-methylpiperidin-1-yl)ethyl)-pyrrolodine-1 sulfonyl)phenol hydrochloride (SB-269970), risperidone, mesulergine and clozapine. In agreement with the decreased binding affinity of 5-HT, 5-CT, RU24969 and 8-OH-DPAT for the variant receptor, these agonists were less potent in stimulating [3H]cAMP accumulation in cells stably expressing the Thr92Lys h5 HT7(a) receptor. Sumatriptan did not stimulate cAMP accumulation in spite of its affinity for both receptor isoforms pointing to a putative weak antagonistic property of this drug at the h5-HT7 receptor. SB-269970 and clozapine were equipotent at both the variant and the wild-type receptor in producing a rightward shift of the 5-HT concentration-response curve for its stimulant effect on [3H]cAMP accumulation. In view of, e.g., the purported involvement of the 5 HT7 receptor in the regulation of circadian rhythm, it may be concluded that the decrease in affinity of 5-HT and other 5-HT receptor agonists at the (Thr92Lys) h5-HT7 receptor may be associated with changes of sleep physiology and of actions of new 5-HT7 receptor agonists designed to treat circadian dysregulation. PMID- 15896882 TI - The protective effects of cystamine in the R6/2 Huntington's disease mouse involve mechanisms other than the inhibition of tissue transglutaminase. AB - Tissue transglutaminase (tTG) is a multifunctional enzyme that contributes to disease progression in mouse models of Huntington's disease (HD), an inherited neurodegenerative disease that shows an age-related onset. Moreover, administration of the transglutaminase inhibitor cystamine delays the onset of pathology in the R6/2 HD mouse model. However, the contribution of tTG inhibition towards the therapeutic effects of cystamine has not been determined, as this compound likely has multiple mechanisms of action in the R6/2 mouse. In this study, we found that administration of cystamine in drinking water delayed the age of onset for motor dysfunction and extended lifespan to a similar extent in R6/2 mice that had a normal genetic complement of tTG compared with R6/2 mice that did not express tTG. Since the magnitude of cystamine's therapeutic effects was not affected by the genetic deletion of tTG, these results suggest that the mechanism of action for cystamine in this HD mouse model involves targets other than tTG inhibition. PMID- 15896883 TI - Systemic mobilization of antigen presenting cells, with a chimeric Flt-3 and G CSF receptor agonist, during immunization of Macaca mulatta with HIV-1 antigens is insufficient to modulate immune responses or vaccine efficacy. AB - In order to improve the efficacy of current vaccine candidates against HIV/AIDS, we sought to strengthen the induction of immune responses via simultaneous in vivo mobilization of dendritic cells using a chimeric Flt-3 and G-CSF receptor agonists (ProGP). We investigated ProGP treatment in combination with two DNA immunizations encoding HIV-Env89.6, SIV-Gag proteins to increase the priming of immune responses. Administration of this Flt-3/G-CSF chimera elicited marked increases in numbers of both plasmacytoid and myeloid dendritic cells. However, there was no increase seen in T-cell responses either directly following the DNA immunization or after further boosting with MVA vectors expressing HIV-Env89.6p, SIV-Gag. After challenge with SHIV89.6p all animals became infected and no differences were seen between the ProGP treated versus the control group with regard to plasma virus load or CD4 T-cell count. We conclude that besides mobilization of dendritic cells additional stimuli to induce dendritic cell maturation may be needed for avid boosting of antigen specific immune activation. PMID- 15896884 TI - The use of the laryngeal mask for tracheostomy. PMID- 15896885 TI - Re: Ganglion cyst of the temporomandibular joint. PMID- 15896886 TI - Purse-string closure of a donor defect in the radial forearm. PMID- 15896887 TI - Review of 509 mandibular fractures treated at the University Hospital, Alexandria, Egypt. AB - Mandibular fracture is one of the most common facial skeletal injuries. Although its main causes are road crashes and violence, the relation between these causes varies from one country to another. We made a retrospective study of the medical records and radiographs of 509 patients treated for mandibular fracture at the University of Alexandria Hospital between 1991 and 2000. The data that we collected included age, sex, aetiology, date of injury, anatomical site of the fracture, associated maxillofacial trauma, and treatment. The prevalence of mandibular fractures was higher in male subjects in all age groups, and the male:female ratio was 3.6:1. Most fractures were sustained by men in the age group 21-30 years and girls between 0 and 10 years, and the monthly incidence was constant. Road crashes were the main cause, followed by falls and assaults. Fractures of the angle were the most common (22%) followed by parasymphyseal fractures (21%) and the lowest was in the coronoid region (1%). PMID- 15896888 TI - Involvement of the jaw bones in systemic mastocytosis. AB - Systemic mastocytosis is characterised by proliferation of mast cells and infiltration of organs. Severe bony pain may result from release of chemical mediators from mast cells and affected patients are at an increased risk of anaphylaxis. Traditional analgesics such as non-steroidal anti-inflammatory agents and opioids are contraindicated. Diagnosis is based on presentation, biopsy of bone marrow, and magnetic resonance imaging of the affected area. In the head and neck, the disease may present as facial pain, localised osteomyelitis, oral sinus formation, and oral ulceration. Treatment is with histamine antagonists and bisphosphonates to control symptoms. An adrenaline pen is provided for use in case of anaphylaxis. PMID- 15896889 TI - Duplication of internal jugular vein and relation to the spinal accessory nerve. AB - We report a case of duplication of the internal jugular vein in which the duplication was incomplete and the accessory nerve lay deep to the vein. PMID- 15896890 TI - The Furlow Z-plasty in two-staged palatal repair modifications and complications. AB - OBJECTIVES: To report the modifications and complications of the Furlow palatoplasty for two-stage closure of the palate. PATIENTS AND METHODS: Prospective study of a consecutive series of 45 primary closures of the soft palate portion of clefts extending into the hard palate; mean (S.D.) age at repair 12 (2) months; median follow-up 4 years 4 months (range 2 months to 9 years). The hard palatal part of the cleft was closed in 18 patients at the mean age of 3 years 11 months. RESULTS: The main modifications that we made were the use of quilting sutures, lateral V-Y closures, and fibrin glue application, and the omission of lateral releasing incisions. Patients stayed in hospital for a median of 4 days (range 3-8 days). Two patients had postoperative partial obstruction of the airway and were given steroids. In six patients, a smaller portion of the oral layer of the wound broke down; it healed by secondary intention in five, but resulted in partial dehiscence in one. There were no oronasal fistulas in the 18 patients who had delayed closure of the hard palate part of the cleft. Secondary pharyngoplasty was not necessary in any patient. CONCLUSION: Furlow's technique has been modified for use in the two-stage closure of complete cleft palates (with or without cleft lip or alveolus) with an acceptable rate of complications. PMID- 15896891 TI - Sensory deficit in the donor hand after harvest of radial forearm free flaps. AB - Fifty patients undergoing radial forearm free flap reconstruction of head and neck defects were examined to find out the extent of sensory defect at the donor site. Flaps (mean length 6 cm, range 4-9) and mean width 4.7 cm (range 3.5-7) were raised. Of the 50 patients 38 (76%) were aware of some sensory loss over the radial distribution in the donor hand. There was objective evidence of a reduction in at least one sensory function in 32 of these patients (84%). The mean affected area was 44.3 cm(2) (range 6-125). The mean length of the affected area was 11.3 cm (range 4.3-12.1) and the mean width 5.1cm (range 2.1-8.4). Of the 12 patients (24%) who reported no feeling of sensory loss all modalities of sensation were preserved in 11 (92%). PMID- 15896892 TI - Taste after reduction of the tongue in Beckwith-Wiedemann syndrome. AB - We tested the sensitivity of taste after reduction of the tongue in four girls with Beckwith-Wiedemann syndrome. No patient had taste blindness, but the ability to detect salty and bitter tastes declined after reduction of the tongue. PMID- 15896893 TI - Monitoring and toxicity of sulfonated derivatives of benzene and naphthalene in municipal sewage treatment plants. AB - Monitoring benzenesulfonates (BS) and naphthalenesulfonates (NS) took place in five municipal sewage treatment plants (STP). A previously optimized method based on solid phase extraction with polymeric cartridges followed by ion-pair liquid chromatography-electrospray-mass spectrometry (SPE-IPC-ESI-MS) was used. This work confirmed the little or no effect of primary settlement on total organic carbon (TOC) and monosulfonated compounds removal, whereas the main reduction is obtained at the biological stage. However, the most polar compounds, such as naphthalenedisulfonates (NDS), were not effectively removed using the biological treatment. An aromatic sulfonated compound is suggested to be used as a tracer of the origin of industrial pollutants discharged into STPs. A bioluminescence inhibition test, Microtox assay, allowed toxicity determination of the most relevant aromatic sulfonated compounds detected and toxicity comparison between primary and secondary effluents. PMID- 15896894 TI - Socio-cultural factors influencing prevention and treatment of tuberculosis in immigrant and Aboriginal communities in Canada. AB - This multi-method study used a participatory action research approach to examine the complex net of socio-cultural factors that influenced behaviour related to tuberculosis (TB) prevention and treatment in the 10 highest risk cultural groups consisting of immigrant and Aboriginal populations in the province of Alberta, Canada. Trained community research associates collected qualitative interview data and helped with interpretation and evaluation. A community advisory committee established foundation principles and monitored the ethical and cultural appropriateness of the research process. A key finding is that although patients with active disease learn about TB from health professionals, people in high-risk populations need to learn more about TB transmission and prevention prior to contact. This is particularly important given that lack of knowledge of TB was strongly associated with negative attitudes towards TB and a worse experience of the disease. The study results underline the need for accessible and culturally appropriate health education about TB in the high risk groups. This can be accomplished in collaboration with lay people, particularly those who have recovered from active TB, their family members and health workers from the community. PMID- 15896895 TI - Limited good and limited vision: multidrug-resistant tuberculosis and global health policy. AB - Almost a third of the world's population is infected with Mycobacterium tuberculosis, the organism that causes tuberculosis disease. Most of those infected never fall ill, but individuals who do can recover if they have access to effective therapies. This paper discusses certain ethical and ethnographic issues raised by cases in which patients are infected with M. tuberculosis strains resistant to at least the two most powerful drugs on which therapy is usually based. In most poor countries, people with such multidrug-resistant tuberculosis (MDR-TB) were, until very recently, considered "untreatable." In addition to being consigned to a permanent state of ill health, they were also at risk of transmitting their resistant strain to others. In this paper we discuss the logic of "cost-effectiveness," which international health policy-makers utilized to make the case that treatment of MDR-TB is not feasible in resource poor settings. These analyses, which have held sway in public health policy for many years, are flawed, we argue, because they ignore and conceal the social determinants of access to health services and often rely on assumptions rather than evidence. We propose that policies based solely on analyses of cost effectiveness of specific interventions for individual settings can be short sighted and, because they do not pay sufficient attention to the social, political, economic, epidemiological and pathophysiological factors influencing the production of health, will ultimately hinder progress toward effective global TB control. PMID- 15896896 TI - [Use of nasolabial flap for mouth floor reconstruction]. AB - SUBJECT: Oral cavity cancers represent 30% of the cephalic extremity tumors. Their resection requires in the majority of the cases a reconstruction by soft tissue. The reconstruction must be simple by bringing some reliable, hairless, thin, resistant tissue to radiation therapy, with a limited morbidity and an acceptable scar ransom. PURPOSE: The purpose of our study is to define the place and the limits of nasolabial flap in the reconstruction of the anterior floor of the mouth after tumoral resection compared to the other surgical and microsurgical techniques. MATERIAL AND METHOD: We retrospectively studied patients with oral cancerous lesions of the anterior floor of the mouth reconstructed by nasolabial flap between 1997 and 2002. The patients benefited from a surgical resection of the tumor by respecting the safety margins, with an immediate reconstruction allowing the restoring of the oral functions. We tried to describe the limits of this flap and discussed its modalities of vascularization. RESULTS: Fifty-three flap procedures were performed on 47 patients; forty-one have received a unilateral and 6 a bilateral nasolabial flap. The majority of tumors were squamous cell carcinomas (50 cases). The average age of patients were 64.8 years (45-78 years) with 40 men (75%) and 13 women (25%). A radical neck lymph nodes dissection with facial artery ligation was realized for 21 patients (15 ipsilateral and 3 bilateral) without affecting the outcome. As complications, we noted one complete necrosis and two partial necrosis of the flap, two postoperative wound complications with dehiscence as well as a massive local recurrence of initial tumor in one patient. CONCLUSION: The nasolabial flap represents a simple functional and morphological option to other pedicled or microsurgically anastomosed flaps for the reconstruction of intermediate-sized mouth floor defects. PMID- 15896897 TI - Recurrent epithelial ovarian cancer: validation and improvement of an established prognostic index. AB - OBJECTIVES: To validate and improve an established prognostic index in patients with recurrent ovarian cancer. METHODS: A Canadian three-covariate prognostic index (tumour grade at diagnosis, initial performance status, and time to relapse/primary progression (TRP)) was validated in a well-defined cohort of comparable Danish patients. Potential parameters to be included in an improved prognostic index were revealed by univariate and multivariate analyses in the Danish validation group. RESULTS: The Canadian index validated in the Danish patient population (n=189) found a statistical significant difference in survival between the prognostic groups good and intermediate (P<0.0001), whereas there was no significant difference in survival between the prognostic groups intermediate and poor (P=0.51). In order to improve the accuracy of the index, the candidate parameters, treatment free interval (TFI), CA125 level and performance status, at time of relapse/primary progression, were added, whereas the parameters, tumour grade, and initial performance status, from the Canadian index were excluded. As the correlation coefficient between TRP and TFI was very high (r=0.91), TRP was substituted with TFI in the improved prognostic model. The final model was: 0.8 (performance status)+0.33 log (CA125)-1.31 log (TFI). The improved model was a good predictor of one-year survival (AUC 0.85; logistic regression; P<0.0001). The median survival (with 95% CI) of the four prognostic groups (A-D) was 50.6 (34.0-not available), 25.0 (22.1-33.6), 11.3 (8.5-12.9), and 5.2 (3.5-6.3) months, respectively. CONCLUSIONS: A novel prognostic model (the Copenhagen index) for patients with recurrent ovarian cancer is presented. PMID- 15896898 TI - The study of bimolecular reactions under non-pseudo-first order conditions. AB - In this work a new equation which describes the time evolution of bimolecular reactions is derived and tested by experiment. The equation is general and the results show that second-order reactions of any simple type may be accurately described by a quotient of exponential functions. The model and reagent concentration dependent observed rate constants show a complex non-linear behaviour when experimental conditions deviate from pseudo-first order nevertheless reducing to the well-known linear dependence when pseudo-first order conditions are met. PMID- 15896899 TI - Influence of the separation of the charged groups and aromatic ring on interaction of tyrosine and phenylalanine analogues and derivatives with beta cyclodextrin. AB - Interactions of tyrosine and phenylalanine analogues with beta-cyclodextrin have been examined in terms of structural features of the ligand such as the separation of the charged amino group and aromatic ring, the presence of additional functional group attached to the amino or phenyl ring, and the presence of a charge on amino or carboxyl group, and steric effects using steady state and time-resolved fluorescence spectroscopy and microcalorimetry. The studied aromatic amino acids possess low binding constant to beta-cyclodextrin, diversified with respect to the presence or absence of a substituent in para position of the phenyl ring. However, calculated, based on the global analysis of the fluorescence intensity decays, binding constants do not allow to estimate unequivocally the influence of the distance between the charged groups and phenol/phenyl ring on the inclusion complex stability because of their low diversification. PMID- 15896900 TI - Computational electrode potential of a coumestan derivative: theoretical and experimental studies. AB - Electrode potential of a coumestan derivative, an important biological molecule, in aqueous solution is computed theoretically using Self-Consistent Field (SCF) theory at the level of Hartree--Fock and employing 6-31G(d) basis set and also obtained experimentally by employing electrochemical technique of cyclic voltammetry (CV). Frequency calculations have been carried out and thermal corrections and entropies have been taken into account. Polarizable continuum model is used to describe the solvent. The theoretical and experimental values for the standard electrode potential of the studied coumestan are in excellent agreement with each other and there is only 0.001 V discrepancy between experiment and theory. The agreement mutually verifies the accuracy of experimental method and the validity of mathematical model. PMID- 15896901 TI - Growth arrest of thyrotropic tumors by thyroid hormone is correlated with novel changes in Wnt-10A. AB - The molecular mechanism underlying thyroid hormone inhibition of thyrotrope cell growth is poorly understood. A comprehensive screen for T3-regulated genes involved in thyrotrope cell regulation was performed by Affymetrix MGU74A Genechip microarray analyses, which compared total RNA from hypothyroid versus 24 h T3-treated TtT-97 tumors. Of the 13,000 genes screened, a number of novel, T3 responsive candidate genes were identified. Within the Wnt family of growth factors, only Wnt-10A transcripts were abundantly expressed in hypothyroid TtT-97 tumors, and were down-regulated with T3 by 6 h of treatment. In addition, nuclear beta-catenin, which is a downstream mediator of canonical Wnt signaling, was decreased at the protein and functional levels. TtT-97 growth suppression was associated with decreased cyclin A transcript levels. We conclude that treatment of thyrotropic TtT-97 tumors with T3 resulted in the decreased expression of Wnt 10A, and that thyroid hormone may inhibit growth via cyclin A regulation. PMID- 15896902 TI - Impact of p53 aberration on the progression of Adult T-cell Leukemia/Lymphoma. AB - Based on statistical analysis of its age-dependent occurrence, a multi-step carcinogenesis model has been proposed for Adult T-cell Leukemia/Lymphoma (ATLL). We have previously reported that the deletion of the p16 gene is a key event in ATLL progression. In the current study, we report for the first time that the aberrations of p16 and p53 are mutually exclusive in ATLL and either of the two events is sufficient for the ATLL progression. More than half of the patients had one of the two aberrations, and both aberrations emerged as significant markers for a poor prognosis. PMID- 15896903 TI - Ontogeny of retrograde labeled chemoafferent neurons in the newborn rat nodose petrosal ganglion complex: an ex vivo preparation. AB - Hypoxic chemosensitivity of the peripheral arterial chemoreceptors in the carotid body is developmentally regulated. Essential neural elements of the chemotransducing unit in the carotid body consist of the Type I cell that depolarizes and releases neurotransmitters in response to hypoxemia and the chemoafferent fibers which form synapses with Type I cells, contain postsynaptic receptors and have cell bodies in the petrosal ganglion. While many properties of the Type I cells have been characterized during postnatal development, less is known about the effect of development on the number and properties of the chemoafferents since localization of the cell bodies of chemoafferents are intermingled with the cell bodies of other sensory neurons that innervate the upper airway. Here, we describe a novel ex vivo preparation that we have developed to retrogradely label cell bodies of chemoafferents in the petrosal ganglion with rhodamine dextran. With this technique, in newborn rats, we show that there is a three-fold increase in retrogradely labeled neurons in the nodose petrosal ganglion complex from postnatal day (PND) 3-7 with a three-fold decrease by PND 14 (P < 0.001, ANOVA). Furthermore, greater than 85% of these retrogradely labeled neurons co-express TH mRNA in all age groups. This novel ex vivo technique circumvents many of the technical difficulties encountered with retrogradely labeling chemoafferents in small newborn animals in vivo, and provides a method to identify and characterize essential neural components of the chemotranductive unit of the peripheral arterial chemoreceptors. PMID- 15896904 TI - The direction of oscillation in spiral drawings can be used to differentiate distal and proximal arm tremor. AB - The assessment of the extent of involvement of the distal and proximal joints in a complex arm tremor is important clinically, as tremor generated around the distal and proximal arm joints can be differentially alleviated by surgical intervention to the thalamic and subthalamic targets, respectively. We observed that the tremor present in spiral drawings shows a diagonal directional preference. We hypothesized that the directional preference of tremor within spiral drawings could be related to the involvement of different arm joints. In this study, we tested this hypothesis by: (1) studying simulated tremulous drawings performed by healthy subjects with the joints selectively restrained; (2) recording surface EMGs from the forearm extensor and deltoid muscles during drawing; and (3) mathematical modelling the kinematics of the arm in 2D drawings. Our data showed that a "main diagonal" axis of orientation of the tremor along the orientation of the forearm indicates a predominantly proximal tremor driven by the shoulder, and a "cross diagonal" axis of orientation of tremor (i.e. perpendicular to the forearm) indicates that the tremor is predominantly distal and driven by the elbow or wrist. We conclude that the spiral drawing task can be used as an objective and quantitative method for differentiating the involvement of the proximal and distal arm joints in complex arm tremors. PMID- 15896905 TI - Cholesteryl ester transfer protein TaqI B and lipoprotein lipase Ser447Ter gene polymorphisms are not associated with ischaemic stroke in Greek patients. AB - Cholesteryl ester transfer protein (CETP) and lipoprotein lipase (LPL) are both key players in plasma lipoprotein homeostasis and, as such, genetically induced alterations in their respective activities may affect susceptibility to cerebrovascular diseases. In this study, we examined the distribution of two common polymorphisms, namely CETP TaqI B and LPL Ser447Ter in a cohort of Greek clinically diagnosed late-onset ischaemic stroke patients (n = 98) and an ethnicity-, age- and sex-matched control group with no manifestations of vascular disease (n = 100). Our study revealed no statistically significant differences with respect to the distribution of either polymorphism, examined separately or in combination, between the two groups. PMID- 15896906 TI - Upregulation of the phosphorylated form of CREB in spinothalamic tract cells following spinal cord injury: relation to central neuropathic pain. AB - Spinal cord injury (SCI) often leads to the generation of chronic intractable neuropathic pain. The mechanisms that lead to chronic central neuropathic pain (CNP) following SCI are not well understood, resulting in ineffective treatments for pain relief. Studies have demonstrated persistent hyperexcitability of dorsal horn neurons which may provide a substrate for CNP. We propose a number of similarities between CNP mechanisms and mechanisms that occur in long-term potentiation, in which hippocampal neurons are hyperexcitable. One biochemical similarity may be activation of the transcription factor, cyclic AMP response element-binding protein (CREB), via phosphorylation (pCREB). The current study was designed to examine whether tactile allodynia that develops in segments rostral to SCI (at-level pain) correlates with an increase in CREB phosphorylation in specific neurons known to be involved in allodynia, the spinothalamic tract (STT) cells. This study determined that, in animals experiencing at-level allodynia 35 days after SCI, pCREB was upregulated in the spinal cord segment rostral to the injury. In addition, pCREB was found to be upregulated specifically in STT cells in the rostral segment 35 days after SCI. These findings suggest one mechanism of maintained central neuropathic pain following SCI involves persistent upregulation of pCREB expression within STT cells. PMID- 15896907 TI - NMDA receptor binding declines differentially in three spinal motor nuclei during postnatal development. AB - The NMDA subtype of glutamate receptors mediates a variety of neuronal processes involved in the development of dendritic morphology. For example, NMDA receptor antagonism during the early postnatal period attenuates dendritic growth in spinal motoneurons. NMDA receptors are present in high levels in the spinal cord early in the postnatal period and decline during development, a period of extensive dendritic plasticity in the spinal cord. Previous studies have suggested that an adult pattern of distribution of NMDA receptors is established as early as postnatal day (P)21 (day of birth = P1). However, given that dendritic growth in spinal motoneurons is not complete by this age and that NMDA receptor activation is necessary for dendritic growth, we assessed NMDA receptor binding in specific spinal motor nuclei during normal development. NMDA receptors were labeled with [3H]MK-801 at P7, P14, P28, P49, and in adult male rats. Receptor binding in the spinal nucleus of the bulbocavernosus (SNB), dorsolateral nucleus (DLN) and retrodorsolateral nucleus (RDLN) was measured using in vitro quantitative autoradiography. NMDA receptor binding over the SNB, DLN and RDLN in intact males was initially high, and declined to adult levels. However, the time course of the decline differed across nuclei. The local decline in NMDA receptor binding observed in the SNB and DLN is coincident with the periods of dendritic growth in these nuclei, further supporting a role for NMDA receptors in the development of motoneuron dendritic morphology. PMID- 15896908 TI - 3D spectrum imaging of multi-wall carbon nanotube coupled pi-surface modes utilising electron energy-loss spectra acquired using a STEM/Enfina system. AB - Numerous studies have utilised electron energy-loss (EEL) spectra acquired in the plasmon (2-10 eV) regime in order to probe delocalised pi-electronic states of multi-wall carbon nanotubes (MWCNTs). Interpretation of electron energy loss (EEL) spectra of MWCNTs in the 2-10 eV regime. Carbon (accepted for publication); Blank et al. J. Appl. Phys. 91 (2002) 1657). In the present contribution, EEL spectra were acquired from a 2D raster defined on a bottle-shaped MWCNT, using a Gatan UHV Enfina system attached to a dedicated scanning transmission electron microscope (STEM). The technique utilised to isolate and sequentially filter each of the volume and surface resonances is described in detail. Utilising a scale for the intensity of a filtered mode enables one to 'see' the distribution of each resonance in the raster. This enables striking 3D resonance-filtered spectrum images (SIs) of pi-collective modes to be observed. Red-shift of the lower energy split pi-surface resonance provides explicit evidence of pi-surface mode coupling predicted for thin graphitic films (Lucas et al. Phys. Rev. B 49 (1994) 2888). Resonance-filtered SIs are also compared to non-filtered SIs with suppressed surface contributions, acquired utilising a displaced collector aperture. The present filtering technique is seen to isolate surface contributions more effectively, and without the significant loss of statistics, associated with the displaced collector aperture mode. Isolation of collective modes utilising 3D resonance-filtered spectrum imaging, demonstrates a valuable method for 'pinpointing' the location of discrete modes in irregularly shaped nanostructures. PMID- 15896909 TI - Chaotropic ions and multivalent ions activate sperm in the viviparous fish guppy Poecilia reticulata. AB - The control system of sperm motility in the fishes with internal fertilization still remains to be studied. In the present study, we examined the sperm activation in the viviparous fish guppy Poecilia reticulata as a function of solute composition. The degree of sperm activation by inorganic and organic solutes correlated well with the lyotropic or Hofmeister series. The solutes that induced sperm activation corresponded to those that have been termed "chaotropes" or macromolecule-destabilizing solutes. In addition to this Hofmeister rule, multivalent cations and anions had valency-dependent activating effects at low concentrations. We investigated the cell signaling triggered by chaotropes and divalent cations and found an intracellular cAMP concentration increase upon the activation of sperm motility. From the present findings, we propose a novel regulation mechanism of sperm motility in the viviparous fish. PMID- 15896910 TI - Risk of morbidity and mortality from cardiovascular disease following radiotherapy for childhood cancer: a systematic review. AB - PURPOSE: To evaluate the existing evidence regarding the long-term risk of cardiovascular disease (CVD) after radiotherapy for childhood cancer. PATIENTS AND METHODS: MEDLINE and EMBASE were searched for articles reporting on radiation induced CVD after childhood cancer published between 1966 and October 2002. Information about study design, population, treatment, outcome and risk factors were abstracted and the quality of each study was assessed. RESULTS: Fourteen articles met all the eligibility criteria. Ten studies evaluated clinical cardiovascular events (CVE) and 11 cardiovascular mortality (CVM) after cardiac irradiation for childhood cancer. Four studies, all in survivors of Hodgkin's disease, showed a significantly increased standardised mortality ratio; a 22- to 68-fold increase compared to the general population. No study compared the risk of CVE with the general population. Three studies examined the risk of CVD (both CVM and CVE) after radiotherapy compared to an unexposed control group, and two showed a significantly increased relative risk. Many studies had important methodological limitations, related to completeness of follow-up, adjustment for other risk factors and outcome assessment in CVE studies. CONCLUSIONS: This systematic review demonstrates that the risk of CVM after cardiac irradiation for childhood cancer is increased compared to the general population and to unexposed patients. The risk of developing clinical CVE and the precise risk factors for developing CVE or CVM after radiotherapy remain unclear. New well-designed studies are needed to reliably evaluate the long-term risk of CVD following radiotherapy and associated risk factors. PMID- 15896911 TI - Lesion-induced differential expression and cell association of Neurocan, Brevican, Versican V1 and V2 in the mouse dorsal root entry zone. AB - Lack of regeneration in the CNS has been attributed to many causes, including the presence of inhibitory molecules such as chondroitin sulfate proteoglycans (CSPGs). However, little is known about the contribution of CSPGs to regeneration failure in vivo, in particular at the dorsal root entry zone (DREZ), a unique CNS region that blocks regeneration of sensory fibers following dorsal root injury without glial scar formation. The goal of the present study was to evaluate the presence, regulation, and cellular identity of the proteoglycans Brevican, Neurocan, Versican V1 and Versican V2 in the DREZ using CSPG-specific antibodies and nucleic acid probes. Brevican and Versican V2 synthesized before the lesion were still present at high levels in the extracellular matrix of the DREZ several weeks after injury. In addition, Brevican was transiently expressed by reactive oligodendrocytes, and by a subset of astrocytes thereafter. Versican V2 mRNA appeared in NG2-positive cells with the morphology of oligodendrocyte precursor cells. Neurocan and Versican V1 levels were low before injury, and appeared in nestin-positive astrocytes and in NG2-positive cells, respectively, following lesion. Versican V1, but not V2, was also transiently increased in the peripheral dorsal root post-lesion. This is the first thorough description of the expression and cell association of individual proteoglycans following dorsal root lesion. It demonstrates that the proteoglycans Brevican, Neurocan, Versican V1, and Versican V2 are abundant in the DREZ at the time regenerating sensory fibers reach the PNS/CNS border and may therefore participate in growth-inhibition in this region. PMID- 15896912 TI - Properties of external plexiform layer interneurons in mouse olfactory bulb slices. AB - In the external plexiform layer (EPL) of the main olfactory bulb, apical dendrites of inhibitory granule cells form large numbers of synapses with mitral and tufted (M/T) cells, which regulate the spread of activity along the M/T cell dendrites. The EPL also contains intrinsic interneurons, the functions of which are unknown. In the present study, recordings were obtained from cell bodies in the EPL of mouse olfactory bulb slices. Biocytin-filling confirmed that the recorded cells included interneurons, tufted cells, and astrocytes. The interneurons had fine, varicose dendrites, and those located superficially bridged the EPL space below several adjacent glomeruli. Interneuron activity was characterized by high frequency spontaneous excitatory postsynaptic potential/currents that were blocked by the alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA)/kainate receptor antagonist 6-cyano-7 nitroquinoxaline-2,3-dione and largely eliminated by the voltage-sensitive Na+ channel blocker, tetrodotoxin. Interneuron activity differed markedly from that of tufted cells, which usually exhibited spontaneous action potential bursts. The interneurons produced few action potentials spontaneously, but often produced them in response to depolarization and/or olfactory nerve (ON) stimulation. The responses to depolarization resembled responses of late- and fast-spiking interneurons found in other cortical regions. The latency and variability of the ON-evoked responses were indicative of polysynaptic input. Interneurons expressing green fluorescent protein under control of the mouse glutamic acid decarboxylase 65 promoter exhibited identical properties, providing evidence that the EPL interneurons are GABAergic. Together, these results suggest that EPL interneurons are excited by M/T cells via AMPA/kainate receptors and may in turn inhibit M/T cells within spatial domains that are topographically related to several adjacent glomeruli. PMID- 15896913 TI - Exercise primes a molecular memory for brain-derived neurotrophic factor protein induction in the rat hippocampus. AB - Exercise is an important facet of behavior that enhances brain health and function. Increased expression of the plasticity molecule brain-derived neurotrophic factor (BDNF) as a response to exercise may be a central factor in exercise-derived benefits to brain function. In rodents, daily wheel-running exercise increases BDNF gene and protein levels in the hippocampus. However, in humans, exercise patterns are generally less rigorous, and rarely follow a daily consistency. The benefit to the brain of intermittent exercise is unknown, and the duration that exercise benefits endure after exercise has ended is unexplored. In this study, BDNF protein expression was used as an index of the hippocampal response to exercise. Both daily exercise and alternating days of exercise increased BDNF protein, and levels progressively increased with longer running duration, even after 3 months of daily exercise. Exercise on alternating days was as effective as daily exercise, even though exercise took place only on half as many days as in the daily regimen. In addition, BDNF protein remained elevated for several days after exercise ceased. Further, after prior exercise experience, a brief second exercise re-exposure insufficient to cause a BDNF change in naive animals, rapidly reinduced BDNF protein to levels normally requiring several weeks of exercise for induction. The protein reinduction occurred with an intervening "rest" period as long as 2 weeks. The rapid reinduction of BDNF by an exercise stimulation protocol that is normally subthreshold in naive animals suggests that exercise primes a molecular memory for BDNF induction. These findings are clinically important because they provide guidelines for optimizing the design of exercise and rehabilitation programs, in order to promote hippocampal function. PMID- 15896914 TI - Elimination of neurokinin-1 receptor neurons in caudal nucleus reverses the effects of systemic bicuculline on c-Fos expression in rat trigeminal sensory nucleus: I. High intensity electrical stimulation of the trigeminal ganglion. AB - Although neurokinin-1 receptor (NK-1)-bearing neurons are distributed in lamina I of the trigeminal caudal nucleus (Vc) and constitute major projection neurons, little is known about their fundamental role(s) in nociceptive processing. This study examines the effect of intra cisterna magna injection of substance P (SP) conjugated to saporin (SP-Sap; 5 microM, 5 microl) [with/without systemic administration of bicuculline] on c-Fos expression in the trigeminal sensory nucleus (TSN) induced 2 h after 10 min repetitive electrical stimulation of the trigeminal ganglion (TG) at high intensity (1.0 mA, 5 Hz, 5 ms) in the urethane anesthetized rat. In the SP-Sap-treated rats, the numbers of NK-1-immunopositive neurons in laminae I and III of the Vc decreased compared with rats similarly pretreated with saline (Sal; 5 microl) or blank-saporin (Bl-Sap; 5 microM, 5 microl). In Sal- or Bl-Sap-treated controls, high intensity stimulation induced c Fos expression in neurons throughout the full extent of ipsilateral superficial layers of the Vc (VcI/II), magnocellular zone of the Vc (VcIII/IV) and the dorsal or dorsomedial subdivisions of the rostral TSN above the obex (trigeminal principal, oral (Vo) and interpolar nuclei). Preadministration of bicuculline (2 mg/kg, i.p.) decreased the numbers of c-Fos-immunopositive neurons in the VcI/II, VcIII/IV and Vo in Sal- or Bl-Sap-treated controls. In contrast, high intensity stimulation induced less c-Fos-immunopositive neurons in the VcI/II and Vo of rats treated with SP-Sap compared with those in Sal- or Bl-Sap-treated controls. In SP-Sap-treated rats preadministered with bicuculline, the numbers of c-Fos immunopositive neurons in the VcI/II and Vo were increased compared with the SP Sap-treated rats preadministered with Sal. These results suggest that NK-1 immunopositive neurons in laminae I and III of Vc play a pivotal role in the nociceptive specific processing in the TSN through GABA(A) receptors. PMID- 15896915 TI - Effects of lead on voltage-gated sodium channels in rat hippocampal CA1 neurons. AB - In this study, the effects of lead (Pb2+) on voltage-gated sodium channel currents (INa) were investigated in acutely dissociated rat hippocampal CA1 neurons using the conventional whole-cell patch-clamp technique. We found that Pb2+ reduced the amplitudes of INa in a concentration-dependent manner, and the effect could be washed out by extracellular application of 3 mM EGTA. The results also showed that at the concentration of 100 microM, Pb2+ decreased the activation threshold and the voltage at which the maximum INa current was evoked and caused negative shifts of INa steady-state activation curve, and enlarged INa tail-currents; Pb2+ induces a left shift of the steady-state inactivation curve, and delayed the recovery of INa from inactivation, and reduced the fraction of available sodium channels; Pb2+ delayed the activation of INa in a concentration- and voltage-dependent manner, and prolonged the time course of the fast inactivation of sodium channels; activity-dependent attenuation of INa was not altered by Pb2+. It was suggested that Pb2+ might exert its effects on sodium channels by binding a specific site on the extracellular side of sodium channels and dragging the IIS4 voltage sensor outwardly. The interaction of Pb2+ with voltage-dependent sodium channels may lead to change in electrical activity and contribute to worsen the neurotoxicological damage. PMID- 15896916 TI - Right atrial stretch activates neurons in autonomic brain regions that project to the rostral ventrolateral medulla in the rat. AB - Activation of the cardiac mechanoreceptors results in changes in sympathetic nerve activity and plays an important role in the responses elicited by elevated blood volume. Stimulation of the reflex influences several key autonomic regions, namely the paraventricular nucleus (PVN), the nucleus of the tractus solitarius (NTS) and the caudal ventrolateral medulla (CVLM). Neurons in these regions project directly to the rostral ventrolateral medulla (RVLM), a critical region in the generation of sympathetic vasomotor tone. The aim of the present experiments was to determine whether neurons in the PVN, NTS and CVLM that are activated by cardiac mechanoreceptor stimulation also project to the RVLM. Animals were prepared, under general anesthesia, by microinjection of a retrogradely transported tracer into the pressor region of the RVLM, and the placement of a balloon-tipped cannula at the junction of the right atrium and the superior vena cava. On the experimental day, in conscious rats, the balloon was inflated to stimulate cardiac mechanoreceptors (n = 9), or left uninflated (control, n = 8). Compared with controls, there was a significantly increased number of Fos-immunoreactive neurons (a marker of activation) in both the PVN (2.5-fold) and NTS (two-fold), but this was not seen in the CVLM. Compared with controls, a significant number of the neurons in the PVN (8%) and NTS (4.0%) that projected to the RVLM were activated. The data suggest that subgroups of RVLM projecting neurons located in the PVN and NTS are involved in the central reflex pathway activated by cardiac mechanoreceptor stimulation. PMID- 15896917 TI - Cortical processing of visceral and somatic stimulation: differentiating pain intensity from unpleasantness. AB - Visceral and somatic pain perception differs in several aspects: poor localization of visceral pain and the ability of visceral pain to be referred to somatic structures. The perception of pain intensity and affect in visceral and somatic pain syndromes is often different, with visceral pain reported as more unpleasant. To determine whether these behavioral differences are due to differences in the central processing of visceral and somatic pain, non-invasive imaging tools are required to examine the neural correlates of visceral and somatic events when the behavior has been isolated and matched for either unpleasantness or pain intensity. In this study we matched the unpleasantness of somatic and visceral sensations and imaged the neural representation of this perception using functional magnetic resonance imaging in 10 healthy right-handed subjects. Each subject received noxious thermal stimuli to the left foot and midline lower back and balloon distension of the rectum while being scanned. Stimuli were matched to the same unpleasantness rating, producing mild-moderate pain intensity for somatic stimuli but an intensity below the pain threshold for the visceral stimuli. Visceral stimuli induced deactivation of the perigenual cingulate bilaterally with a relatively greater activation of the right anterior insula-i.e. regions encoding affect. Somatic pain induced left dorso-lateral pre frontal cortex and bilateral inferior parietal cortex activation i.e. regions encoding spatial orientation and assessing perceptual valence of the stimulus. We believe that the observed patterns of activation represent the differences in cortical process of interoceptive (visceral) and exteroceptive (somatic) stimuli when matched for unpleasantness. PMID- 15896918 TI - Dexamethasone suppression test in borderline personality disorder--effects of posttraumatic stress disorder. AB - BACKGROUND: Divergent findings of hypothalamic-pituitary-adrenal (HPA) axis functioning in borderline personality disorder (BPD) may be caused by a different degree of comorbid posttraumatic stress disorder (PTSD), in which alterations of the HPA axis are well known. Here we investigate alterations of the HPA axis in BPD patients with and without comorbid PTSD compared to healthy controls. Considering previous findings current major depression (MDD) was taken into account as a confounding variable. METHODS: Apart from clinical assessment the 0.5 mg dexamethasone suppression test (DST) was performed in 21 female borderline patients and 23 healthy controls. RESULTS: Twelve BPD patients suffered from comorbid PTSD. Relative suppression (%) did not differ between healthy controls and the total BPD group, but BPD patients with comorbid PTSD showed increased suppression compared to those without. Comorbid MDD was not associated with suppression. CONCLUSIONS: Our results do not indicate a dysfunction of the HPA axis in BPD. However, comorbid PTSD seems to be associated with a relative hypersuppression in the 0.5 mg DST. PMID- 15896919 TI - Interaction of brain noradrenergic system and the hypothalamic-pituitary-adrenal (HPA) axis in man. AB - BACKGROUND: Numerous interactions between the brainstem locus coeruleus system and the HPA axis have been shown in experimental animals. This relationship is less well characterized in humans and little is known about the influence of psychiatric disorders, which disturb one of these systems, on this relationship. METHODS: Untreated subjects with pure MDD (n = 13), MDD with comorbid anxiety disorders (n = 17), and pure anxiety disorders (n = 15) were recruited by advertising. Age and sex matched control subjects were recruited for each subject with a psychiatric diagnosis (n = 45). All subjects underwent a social stressor, the Trier Social Stress Test (TSST), and blood was collected for ACTH assay. These same subjects also underwent a clonidine challenge study for assessment of growth hormone release as a marker of tonic noradrenergic activation. RESULTS: Examining log transformed area under the curve response for each hormone, a significant negative relationship (simple regression) was observed between systems in normal subjects. This relationship was preserved in anxiety subjects. However, both pure depressed and comorbid depressed and anxiety subjects demonstrated disruption of this relationship. CONCLUSIONS: Under normal circumstances, noradrenergic systems can influence the magnitude of the HPA axis response to stress. However, in subjects with major depression, HPA axis activation appears autonomous of noradrenergic influence. PMID- 15896920 TI - Effects of psycho-social stress during pregnancy on neuroendocrine and behavioural parameters in lactation depend on the genetically determined stress vulnerability. AB - The neuroendocrine consequences of repeated exposure of the pregnant mother to relevant stressors have been studied in the offspring, but not in the mothers. As these stress effects might depend on the genetically determined stress susceptibility of the dams, here, we investigated the effects of daily exposure to psycho-social stressors (maternal defeat by an aggressive lactating resident and restraint) between pregnancy days 4 and 18 in female rats selectively and bidirectionally bred for high (HAB) or low (LAB) anxiety-related behaviour. ACTH and corticosterone secretory responses to a mild stressor were found to be low in unstressed lactating HAB and LAB dams (day 8 of lactation) indicating an intact physiological attenuation of the HPA axis at this time. Pregnancy stress significantly increased the reactivity of the hypothalamo-pituitary-adrenal (HPA) axis in lactating HAB, but not LAB rats, reflecting impaired attenuation of the HPA axis selectively in pregnancy-stressed HAB dams. The high and low anxiety phenotypes were consistent in lactation and not significantly altered by pregnancy stress, despite an elevated level of arousal in pregnancy-stressed HAB dams. In general, HAB dams showed signs of a more protective maternal behaviour compared to LAB dams: (i) in the home cage, HAB dams spent more time in direct pup contact (day 1 of lactation), (ii) during two forms of the pup retrieval test, differing in the level of challenging the dam, HAB dams retrieved the pups faster, and (iii) during the maternal defence test, they were more aggressive towards a virgin intruder compared to LAB and NAB dams. Pregnancy stress did not alter any of these behavioural measures, except an increase in the speed of pup collection in a novel environment in HAB dams and increased maternal aggression in LAB dams. The results indicate a robust behavioural phenotype of HAB and LAB dams with respect to anxiety and maternal behaviour which was found to be almost unchanged by exposure to pregnancy stress. However, the finding of differential effects of pregnancy stress on the attenuation of the reactivity of the HPA axis in lactation makes HAB and LAB rats a potential animal model for studying genetically determined differences in stress vulnerability and stress-induced maladaptation of the HPA axis post-partum. PMID- 15896921 TI - Unaided smoking cessation and predictors of failure to quit in a community sample: effects of gender. AB - The majority of smokers who eventually quit do so without formal treatment, but few studies have examined a range of predictors of cessation among self-quitters and whether they differ depending on smokers' gender. The present study examined predictors of quitting in a community sample of 93 men and 117 women smokers attempting to quit on their own. Smokers were mailed a baseline questionnaire battery, and instructed to complete other measures the first and second day of their quit attempt. No gender difference in quit rates was observed, but predictors of smoking differed between men and women. Among men, alcohol use, caffeine consumption, and negative affect predicted smoking on quit day. Among women, nicotine dependence, age, ethnicity, weight control smoking, and not eating during urges predicted smoking on quit day. Social support seeking was negatively associated with smoking on the quit day among women, but the opposite relationship was obtained for men. Men's smoking 4 months later was predicted by baseline social pressures to quit and use of nicotine replacement therapy (NRT). For women, weight control smoking and trait anxiety predicted smoking at the 4 month follow-up. PMID- 15896922 TI - Retrospective reports of parenting received in their families of origin: relationships to adult attachment in adult children of alcoholics. AB - The present study examined general and romantic attachment and parenting students received in their families of origin among 401 college students who resided with an alcohol-abusing parent prior to age 16 years as compared to those who did not reside with alcohol-abusing parents. Participants completed the Children's Report of Parent Behavior Instrument [Schludermann, E. and Schludermann, S. (1970). Children's Report of Parent Behavior Inventory (CRPBI). Canada: University of Manitoba], Experiences in Close Relationships--Revised [Fraley, R. C., Waller, N. G., and Brennan, K. G. (2000). An item response theory analysis of self-report measures of adult attachment. Journal of Personality and Social Psychology, 78, 350-365], Relationship Scale Questionnaire [Griffin, D. W. and Bartholomew, K. (1994). Models of the self and other: Fundamental dimensions underlying measures of adult attachment. Journal of Personality and Social Psychology, 67, 430-445], and the Children of Alcoholics Screening Test [Jones, J. W. (1983). The Children of Alcoholics Screening Test: Test manual. Chicago: Camelot]. Young adults who met criteria for ACOAs reported more anxious and avoidant behavior in romantic relationships and a more fearful style of general adult attachment. Parenting behavior in one's family of origin predicted anxious behavior in romantic relationships and a fearful overall style of attachment, whereas being an ACOA and parenting in one's family of origin predicted avoidant behavior in romantic relationships. PMID- 15896923 TI - Autoimmune response to anti-apoptotic protein survivin and its association with antibodies to p53 and c-myc in cancer detection. AB - Survivin, an inhibitor of apoptotic protein, is over-expressed in many cancers but not in normal differentiated adult tissues. Recently, antibodies to survivin have been demonstrated in patients with lung and colorectal cancer. Whether antibodies to survivin can be used as a marker for the diagnosis of cancer, and how antibody to survivin is related to antibodies against tumor suppressor protein p53 and oncoprotein c-myc remains to be evaluated. In the present study, the full-length recombinant proteins survivin, p53 and c-myc, were expressed and used as antigens in enzyme-linked immunosorbent assay (ELISA) and Western blot for the detection of antibodies to these three proteins. Sera from 1137 patients with 11 different types of cancer were analyzed. Antibodies to survivin were detected in 8.4% (96/1137), with a significant difference from the control groups consisting of normal individuals and autoimmune disease patients (p<0.05). Of 1137 cancer sera, 546 were also tested for the presence of antibodies to p53 and c-myc. Frequencies of antibodies to p53 and c-myc were 11.5 and 12.3%, respectively. Although antibodies to either one of three antigens do not reach levels of sensitivity, which could become routinely useful in diagnosis, it appears that there are different patterns of antibody frequency in individual cancer type. The results also indicated that when the presence of antibody to any one of these three antigens was considered, the cumulative frequency was increased to 27.3% (149/546) for the total group of cancer patients. It became apparent from our data that the combination of antibodies might acquire higher sensitivity. PMID- 15896924 TI - X-ray repair cross-complementing group 1 polymorphisms and cancer risks in Asian populations: a mini review. AB - X-ray repair cross-complementing group 1 (XRCC1) is an important DNA repair protein. Arg194Trp, Arg280His, and Arg399Gln are three polymorphisms of XRCC1 that commonly exist in human. In this context, we obtained the relevant articles through a PubMed search and examined the association of XRCC1 polymorphisms and the risk of cancer in Asian populations. Generally, a single XRCC1 polymorphism is weakly associated with cancer in Asian populations. However, when combined with other genetic polymorphisms or such lifestyle factors as smoking, XRCC1 polymorphisms show a stronger association with the risk of cancer. The interaction of the 399Gln/Gln genotype and smoking might be associated with a three-fold increase in the risk of cancer. In this paper we provide some important information for practical future cancer prevention programs. To further clarify the association of XRCC1 polymorphisms and cancer risks, additional studies are required from the perspectives of epidemiology and in vitro. PMID- 15896925 TI - Association between subject factors and colorectal cancer screening participation in Ontario, Canada. AB - Colorectal cancer screening reduces colorectal cancer incidence and mortality. This population-based study was conducted to evaluate (i) the association between subject factors and colorectal screening participation and (ii) the lifetime prevalence of colorectal screening among the general population of Ontario, Canada. Population-based controls were recruited by the Ontario Familial Colorectal Cancer Registry during 1998-2000. The 1944 persons completed an epidemiologic questionnaire. Descriptive statistics were computed and step-wise multivariate logistic regression was used to estimate odds ratios and 95% confidence intervals. Overall, 23% of persons greater than 50 years of age reported ever having had colorectal cancer screening; 17% reported fecal occult blood test (FOBT), 6% sigmoidoscopy, and 4% colonoscopy. Family history of colorectal cancer, increased age, higher household income, and use of hormone replacement therapy (among women) were all significantly associated with ever having had colorectal cancer screening. The low prevalence of colorectal cancer screening among the target population suggests the need for an increased awareness of the public health importance of colorectal cancer screening. PMID- 15896926 TI - Antibacterial activity of the essential oil from Cymbopogon nervatus inflorescence. AB - Antibacterial activity of essential oil of dried inflorescence of Cymbopogon nervatus was investigated. The essential oil remarkably inhibited the growth of tested bacteria except for Salmonella typhi. The maximum activity was against Shigella dysenteriae and Klebsiella pneumoniae. PMID- 15896927 TI - Diagnostic reliability of the Semi-structured Assessment for Drug Dependence and Alcoholism (SSADDA). AB - The Semi-structured Assessment for Drug Dependence and Alcoholism (SSADDA) is a diagnostic instrument developed for studies of the genetics of substance use and associated disorders. The SSADDA provides more detailed coverage of specific drug use disorders, particularly cocaine and opioid dependence, than existing psychiatric diagnostic instruments. A computerized version of the SSADDA was developed to permit direct entry of subject responses by the interviewer. This study examines the diagnostic reliability of the SSADDA for substance use disorders and for other DSM-IV disorders that are commonly associated with substance use disorders. METHODS: Two hundred and ninety-three subjects (mean age = 39 yr, 52.2% women) were interviewed twice over a 2-week period in two sub studies examining the inter-rater (n = 173) or test-retest reliability (n = 120) of the SSADDA. The kappa statistic and Yule's Y were used to measure reliability. RESULTS: The reliability of most substance dependence diagnoses was good to excellent, although the reliability of substance abuse diagnoses was substantially lower. The reliability of the associated psychiatric diagnoses varied from fair to excellent. CONCLUSIONS: The SSADDA yields reliable diagnoses for a variety of psychiatric disorders, including alcohol and drug dependence. Although developed for use in genetic studies, its broad and detailed coverage of disorders and computer-assisted format will allow it to be used in a variety of applications requiring careful diagnostic assessment. PMID- 15896928 TI - Adolescent substance use and hospital presentations: a record linkage assessment of 12-month outcomes. AB - AIMS: To examine the prevalence of different substances used by adolescents admitted to hospital emergency departments (ED); to evaluate the impact of an ED based brief intervention (BI) on hospital events; to compare outcomes for those using "alcohol alone", "alcohol plus illicit+/-licit drugs" ("alcohol plus"), or "other drugs" excluding alcohol, and investigate the relationship between hazardous alcohol consumption patterns and hospital events. DESIGN: We used hospital record linkage to follow-up a randomised control trial cohort. PARTICIPANTS: Adolescents (12-19 years) recruited in ED with presentations involving alcohol or other drugs (AOD): 67 received usual care and 60 a BI that facilitated attendance at community drug agencies. MEASUREMENTS: Drug-use categories were assigned from the substances used at the baseline presentation. Outcomes were assessed as hospital admissions plus ED presentations in the 12 month post-intervention. "Hazardous" alcohol use was categorised via the AUDIT-3. RESULTS: The drug-use categories were "alcohol alone" (n=67, 53%), "alcohol plus" (n=31, 24%) and "other drugs" (n=28, 22%). In the 12-month post-intervention, the randomisation groups had similar numbers of AOD hospital events. A Cox regression showed that in the usual care but not the BI group, for "other drugs" there was a 8-fold increased hazard ("risk") of an AOD hospital event compared with "alcohol alone" and a 10-fold increase compared to 'alcohol plus'. Each pre-recruitment AOD event doubled the hazard of an AOD event. For the BI group, these were not significant predictors. The "other drugs" group had more AOD events than either of the other groups. "Hazardous" (77%) alcohol use was common but was not a predictor of AOD hospital events. CONCLUSIONS: BI can be delivered in ED and reduce hospital AOD morbidity associated with the use of drugs other than alcohol. Interventions should focus on those with prior AOD events and "other drugs" presentations. PMID- 15896929 TI - Screening for alcohol problems in two emergency service samples in Poland: comparison of the RAPS4, CAGE and AUDIT. AB - Prior research on alcohol-related problems among emergency service patients in Poland found substantial alcohol involvement on the part of injured males, suggesting emergency services may be a productive venue for identifying patients who could benefit from a brief intervention or referral for treatment. Performance of the RAPS4, CAGE and AUDIT against ICD-10 and DSM-IV criteria for alcohol dependence and for alcohol abuse/harmful drinking was compared in probability samples of emergency service patients from two regions of Poland. Sensitivity of the RAPS4 and AUDIT was significantly better than the CAGE for alcohol dependence among males in Warsaw, but specificity was poorer. Among females, although numbers were small, sensitivity for alcohol abuse/harmful drinking and for alcohol dependence or abuse/harmful drinking was significantly better for the RAPS4-QF than for the CAGE or AUDIT at a cut point of 8 across both sites. Performance of the AUDIT at a cut point of 3 was similar to the RAPS4 QF for females. Among males, sensitivity was higher but specificity considerably lower for the RAPS4-QF compared to the CAGE at a cut point of 1 or for the AUDIT at a cut point of 8. Alternate cut points for the AUDIT optimized performance. Findings suggest some regional and gender differences in performance of screening instruments in these Polish samples, but no instrument or cut point is optimal in identifying those with alcohol use disorders. Additional cross-cultural research is needed to evaluate the performance of instruments, especially among females with alcohol use disorders. PMID- 15896930 TI - Retrograde regulation of multidrug resistance in Saccharomyces cerevisiae. AB - Communication between the mitochondria and the nucleus is essential to ensure correct metabolic coordination of the cell. Signaling pathways leading from the mitochondria to the nucleus are referred to as retrograde signaling and were first discovered in the yeast Saccharomyces cerevisiae. Cells that lack their mitochondrial genome (rho0 cells) trigger expression of the nuclear CIT2 gene in order to ensure adequate amino acid biosynthesis. More recently, it has been found that a different set of genes involved in multidrug resistance in S. cerevisiae is strongly induced in rho0 cells. During a search for negative regulators of the ATP-binding cassette (ABC) transporter-encoding gene PDR5, it was observed that rho0 mutants exhibited dramatic up-regulation of the transcript of this gene. This induction was due to the post-translational activation of a direct upstream regulator of PDR5 that was designated Pdr3p. Loss of the LGE1 gene led to a block in rho0-mediated induction of PDR5 expression. Lge1p has been observed by others to be involved in histone H2B ubiquitination along with the ubiquitin-conjugating enzyme Rad6p and the ubiquitin ligase Bre1p. Our studies provide evidence that Lge1p has another function unique from H2B ubiquitination that is required for retrograde regulation of PDR5 transcription. We have also found that the Pdr pathway regulates expression of several genes involved in sphingolipid biosynthesis. These findings suggest that the physiological role of the PDR genes might be to regulate membrane homeostasis and rho0-triggered changes in this parameter may be the signal controlling PDR gene expression. PMID- 15896931 TI - Comparison of the effect of natural mating, LH, and GnRH on interval to ovulation and luteal function in llamas. AB - Gonadotropins and GnRH have been used to electively induce ovulation in llamas and alpacas, but critical evaluation of the natural interval to ovulation after mating has not been performed nor has a direct comparison of the effects of natural mating versus hormone treatments on this interval and subsequent luteal development. The objectives of this study were to compare the effects of hormonal treatments and natural mating on ovulation induction, interval to ovulation, and luteal development in llamas. The ovaries of llamas were examined by transrectal ultrasonography once daily. Llamas with a large follicle were assigned randomly to be: (1) mated with an intact male (mated; n=10); (2) given 5 mg of LH im (LH; n=11); or (3) 50 microg of GnRH im (GnRH; n=10). Ultrasound examinations were performed every 4h from treatment (day 0) to ovulation and thereafter once daily for 15 consecutive days to monitor CL growth and regression (n=5 per group). Plasma progesterone concentrations were measured at days 0, 3, 6, 9, and 12 after treatment to evaluate CL function. The size of the largest preovulatory follicle at the time of treatment did not differ among groups (11+/-0.6, 10.5+/-0.8, 11.8+/-0.9 mm, for mated, LH, and GnRH groups, respectively; P=0.6). No differences were detected among groups (mated, LH, and GnRH) in ovulation rate (80%, 91%, 80%, respectively; P=0.6), or interval from treatment to ovulation (30.0+/-0.5, 29.3+/-0.6, 29.3+/-0.7h, respectively; P=0.9). Similarly, no differences were detected among groups (mated, LH, and GnRH) in maximum CL diameter (14.2+/-0.3, 13.2+/-0.5, and 13.0+/-0.7 mm, respectively; P=0.5), the day of maximum CL diameter (7.6+/-0.2, 7.6+/-0.2, and 7.4+/-0.4 mm, respectively; P=0.6), or the day on which the CL began to regress (12.3+/-0.3 [non-pregnant, n=3], 11.8+/-0.6, 12.2+/-0.4, respectively; P=0.4). The diameter of the CL and plasma progesterone concentrations changed over days (P<0.0001) but the profiles did not differ among groups. In summary, ovulation rate, interval to ovulation, and luteal development were similar among llamas that were mated naturally or treated with LH or GnRH. We conclude that both hormonal preparations are equally reliable for inducing ovulation and suitable for synchronization for artificial insemination or embryo transfer program. PMID- 15896932 TI - Effect of cholesterol and ethanol on dermal delivery from DPPC liposomes. AB - The main objective of the present work was to compare the dermal delivery of minoxidil (Mx), a lipophilic drug from ethosomes versus classic liposomes, containing different cholesterol (CHOL) concentrations. All the systems were characterized for shape, lamellarity, particle size and entrapment efficiency percentage (EE), by transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), laser diffraction and ultracentrifugation or dialysis methods, respectively. Multilamellar vesicles (MLVs) were obtained and one to six lamellae were visualized by CLSM. The presence of ethanol in the formulations affects the particle size in terms of reducing this parameter. In addition, it was possible to appreciate the influence of CHOL on the vesicle size, because it was increased, as CHOL concentration was higher. When the EE was determined by two different methods (ultracentrifugation and dialysis methods), a clear losing of entrapped drug by the ultracentrifugation method was observed, because the strong energy transmitted to the samples disrupted vesicles. Vesicles were non occlusively applied on rat skin and the permeation pattern of the different systems, depth into the skin and the main permeation pathway were studied by using beta-carotene as a fluorescent probe. CLSM studies showed that ethosomal systems were much more efficient at delivering the fluorescent substance into the skin in terms of quantity and depth, than either liposomes or hydroalcoholic solutions. PMID- 15896933 TI - Colloidal carriers and blood-brain barrier (BBB) translocation: a way to deliver drugs to the brain? AB - The major problem in drug delivery to the brain is the presence of the blood brain barrier (BBB) which limits drug penetration even if in certain pathological situations the BBB is partly disrupted. Therefore, various strategies have been proposed to improve the delivery of drugs to this tissue. This review presents the status of the BBB in healthy patients and in pathologies like neurodegenerative, cerebrovascular and inflammatory diseases. The second part of this article aims to review the invasive and non-invasive strategies developed to circumvent the BBB and deliver drugs into the brain. The use of nanotechnologies (liposomes, nanoparticles) is especially discussed in the ultimate part of the review evidencing their potentiality as non-invasive technique in the brain delivery of drugs with the possibility to target specific brain tissue thanks to ligand linked to carrier surface. PMID- 15896934 TI - Chemical composition and antibacterial activity of essential oil of Pulicaria odora L. AB - The chemical composition of the volatile oil constituent from Pulicaria odora L. roots has been analyzed by GC/MS. Twenty-seven components were identified, being thymol (47.83%) and its derivative isobutyrate (30.05%) the main constituents in the oil. Furthermore, the oil was tested against seven bacteria at different concentrations. Results showed that the oil exhibited a significant antibacterial activity. PMID- 15896935 TI - Thujone--cause of absinthism? AB - Habitual abuse of the wormwood spirit absinthe was described in the 19th and 20th centuries as a cause for the mental disorder "absinthism" including the symptoms hallucinations, sleeplessness and convulsions. A controversial discussion is going on if thujone, a characteristic component of the essential oil of the wormwood plant Artemisia absinthium L., is responsible for absinthism, or if it was merely caused by chronic alcohol intoxication or by other reasons such as food adulterations. To ascertain if thujone may have caused absinthism, absinthes were produced according to historic recipes of the 19th century. Commercial wormwood herbs of two different manufacturers, as well as self-cultivated ones, were used in a concentration of 6 kg/100 l spirit. In addition, an authentic vintage Pernod absinthe from Tarragona (1930), and two absinthes from traditional small distilleries of the Swiss Val-de-Travers were evaluated. A GC-MS procedure was applied for the analysis of alpha- and beta-thujone with cyclodecanone as internal standard. The method was shown to be sensitive with a LOD of 0.08 mg/l. The precision was between 1.6 and 2.3%, linearity was obtained from 0.1 to 40 mg/l (r = 1.000). After the recent annulment of the absinthe prohibition all analysed products showed a thujone concentration below the maximum limit of 35 mg/l, including the absinthes produced according to historic recipes, which did not contain any detectable or only relatively low concentrations of thujone (mean: 1.3 +/- 1.6 mg/l, range: 0-4.3 mg/l). Interestingly, the vintage absinthe also showed a relatively low thujone concentration of 1.8 mg/l. The Val-de Travers absinthes contained 9.4 and 1.7 mg/l of thujone. In conclusion, thujone concentrations as high as 260 mg/l, reported in the 19th century, cannot be confirmed by our study. With regard to their thujone concentrations, the hallucinogenic potential of vintage absinthes can be assessed being rather low because the historic products also comply with today's maximum limits derived to exclude such effects. It may be deduced that thujone plays none, or only a minor role in the clinical picture of absinthism. PMID- 15896936 TI - Development of multiplex PCRs for evolutionary and forensic applications of 37 human Y chromosome SNPs. AB - This work describes an efficient and rapid test for typing 37 single nucleotide polymorphisms (SNPs) of the non-recombining region of Y chromosome (NRY) from a minimal amount of DNA using six PCR multiplexes. Markers were drawn following a hierarchical strategy based on the phylogenetic tree of Y chromosome proposed by the Y Chromosome Consortium [The Y Chromosome Consortium, A nomenclature system for the tree of human Y-chromosomal binary haplogroups, Genome Res. 12 (2002) 339 348]. Two multiplexes--arbitrarily named MY1 and MY2--were developed to explore the basal branches of the tree encompassing all the major clades A-R: MY1 for markers M35, M89, M172, M170, M9, M173, M45 and MY2 for markers M52, M216, M174, M181, M201, M91, M96, M214. Four multiplexes able of typing the more superficial branches typical of most frequent European haplogroups E3b, J2, R1 and I, were also developed and named MY-E3b (M78, M107, M224, M165, M148, M81), MY-J2 (M158, M68, M47, M102, M137, M67), MY-R1 (M17, M269, M18, P25, SRY10831.2) and MY-I (M72, M223, M26, M21, M161). SNP genotyping was carried out by hot-start PCR amplification with primers yielding fragments between 63 and 210 nucleotides, followed by minisequencing reaction based on dideoxy single-base extension and capillary electrophoresis of extension products. The sequential application of these multiplexes is a robust and effective resource for typing the most frequent European Y-SNP haplogroups, and appears to be suitable for forensic purposes and evolutionary studies. PMID- 15896937 TI - Keeping the pieces together: comparison of mass grave excavation methodology. AB - Mass graves are a complex and confusing mix of bodies, body parts, soils, artifacts, and other feature evidence. Forensic investigations of these complex crime scenes should attempt to maximize the collection of evidence, which includes the mortal remains, in their best possible condition as they were deposited within the graves. Two standard methods of mass grave excavation were examined with the aim of identifying the better approach. Two experienced teams using different methodologies excavated two separate but very comparable mass graves located in the same area. Single disarticulated skeletal elements not associated with a body at the time of removal from the grave were categorized and their counts analyzed to evaluate the efficiency of the differing excavation methodologies. The methodologies used were the 'pedestal' method, which focuses on the body mass, and the 'stratigraphic' method, in which the grave feature and contents are conjointly excavated. The first grave (Grave A), excavated using the 'pedestal' method, was observed to have a disproportionately larger amount of unassociated bones than did the second (Grave B), which used the 'stratigraphic' method. Chi-square (chi2) goodness-of-fit and contingency tests were performed on the total numbers of recorded elements and different categorical groups of bones, based on size and shape, in each grave. Results demonstrate that significantly greater numbers of unassociated elements resulted from the excavation of Grave A using the pedestal method, both in total number of disarticulated bones as well as within 'large' and 'medium' categories. Conversely 'small' skeletal elements were recovered at a higher rate in Grave B. The lower 'large' and 'medium' bone production rates from Grave B indicate that the stratigraphic method better maintained the provenience and articulation of remains than did Grave A, while the higher 'small' bone recovery rate may point to better recovery techniques of Grave B's excavation team. PMID- 15896938 TI - Signal intensity, clinical activity and cross-sectional areas on MRI scans in thyroid eye disease. AB - The signal intensity from inflamed extra-ocular muscles on short tau inversion recovery (STIR)-sequence magnetic resonance imaging (MRI) is known to correlate with clinical scores of thyroid eye disease (TED) severity. Twenty-one patients who had undergone repeated MRI scanning for TED were studied retrospectively. Signal intensity of extra-ocular muscles (from STIR-sequence MRI) and cross sectional area (from STIR and T1 MRI) were correlated with Mourits' clinical activity score (CAS). The area of highest signal intensity within the most inflamed extra-ocular muscle, and the average cross-sectional signal intensity of the most inflamed extra-ocular muscle reliably correlated with CAS, and this was maintained as disease activity changed over time. In contrast, isolated measures of muscle cross-sectional area did not correlate with CAS. The extra-ocular muscle cross-sectional area calculated from STIR-sequence MR images was greater than that measured on T1 images. This suggests that muscle area from STIR sequence MRI may also detect peri-muscular inflammation. We conclude that the peak signal intensity from the most inflamed extra-ocular muscle remains the most reliable correlate of clinical disease activity obtained from these images. STIR sequence MRI scans provide a number of useful measures of disease activity in TED. PMID- 15896939 TI - A disposable electrochemical sensor for the rapid determination of levodopa. AB - Levodopa (L-dopa), the biological precursor of catecholamines, is the most widely prescribed drug in the treatment of Parkinson's disease. The present work presents a proposal for the application of a gold screen-printed electrode an electrochemical sensor for monitoring L-dopa in stationary solution and a flow system. Using the electrooxidation of L-dopa at +0.63 V in acetate buffer pH 3.0 on a gold screen-printed electrode it is possible to obtain a linear calibration curve from 9.9 x 10(-5) to 1.2 x 10(-3) mol L(-1) and a detection limit of 6.8 x 10(-5) mol L(-1). Under amperometric conditions (E(app) = 0.8 V; flow rate = 14.1 mL min(-1); pH 3.0), an analytical calibration graph for l-dopa was obtained from 1.0 x 10(-6) mol L(-1) 6.6 x 10(-4) mol L(-1) with a detection limit of 9.9 x 10( 7) mol L(-1). The method was successfully applied to the determination of L-dopa in commercial dosage forms without any pre-treatment. PMID- 15896940 TI - Quantitative determination of ketoprofen in gels and ampules by using flow injection UV spectrophotometry and HPLC. AB - A flow-injection UV spectrophotometric method for the determination of ketoprofen in gels and ampules was developed. Quantitative determination of ketoprofen was realized by using distilled water as a carrier for gels and citrate buffer, pH 6.5, for ampules at 261 nm. No spectrophotometric interferences from additives of gels, carboxypolymethylene and triethanolamine, were observed. There were also no spectrophotometric interferences resulting from additives of ampules named as benzyl alcohol and arginine. The detection limits were 0.436 and 0.303 microg/ml for gels and ampules, respectively. Throughout the study, the flow rate, loop volume and the number of injection per hour were 13.8 mlmin(-1), 193 mcirol and 85, respectively. Analytical signal of the ketoprofen was linear in the concentration range of 7.5-75 microg/ml. Quantitative results of ketoprofen in gels, 25.25+/-0.27 (mean+/-S.D.), and in ampules, 99.42+/-0.44 were in good agreement with the labeled quantities (25 mg/1g gel, 100 mg/2 ml ampule). The recoveries were in the range of 98.65-100.63 and 99.1-101.5% for gels and ampules, respectively. Results obtained were in accordance with those obtained by HPLC. It was seen that the proposed method was fast, accurate, precise and suitable for automation as an analytical method. PMID- 15896941 TI - Early detection of bone infection and differentiation from post-surgical inflammation using 2-deoxy-2-[18F]-fluoro-D-glucose positron emission tomography (FDG-PET) in an animal model. AB - Diagnosing bone infection in the context of post-surgical inflammation is problematic since many of the early signs of infection are similar to normal post surgical changes. We used a rabbit osteomyelitis model to evaluate the use of 2 deoxy-2-[(18)F]-fluoro-d-glucose positron emission tomography (FDG-PET) as a means of detecting post-operative infection in the context of post-surgical inflammation. Comparisons were made between infected and non-infected rabbits in which infection with Staphylococcus aureus was initiated at the time of surgery. Weekly PET scans were obtained 30 and 60 min after the introduction of FDG and analyzed based on standardized uptake values (SUV) at the surgical site and visual assessment of the presence or absence of infection. Concurrent X-rays were taken immediately prior to scanning. At 4weeks post-operatively, animals were sacrificed for histologic and bacteriologic confirmation of infection. Uptake of FDG was evident in the bone of all rabbits on day 1 post-surgery, however, SUV comparisons from the surgical site could not be used to distinguish between the infected and uninfected groups until day 15. Visual analysis of FDG-PET scans revealed a significant difference (p<0.01) between the infected and uninfected groups as early as day 8. This was due in part to the ability to visualize regional lymph nodes by FDG-PET. PMID- 15896942 TI - Individual patient's preferences for hypertension management: a Q-methodological approach. AB - OBJECTIVE: To systematically explore and elicit individual patient's preferences in the management of their hypertension using Q-methodology. METHODS: Using Q methodology, 120 patients ranking 42 statements according to their agreement or disagreement when considering appropriate hypertension management. The statements were derived from an earlier qualitative study. Factor analysis of the data was undertaken using PQMethod software to determine if any patterns were discernible. RESULTS: Ninety-two patients clustered to five factors, which all varied in the degree of involvement patients had, or wished to have, in their hypertension management. The 42 patients who loaded to factor 1 considered that appropriate antihypertensive treatment involved leaving medical decisions to their GPs and trusting their judgement in such matters. The patients (n = 31) who positively loaded to factor 2 suggested that an autonomous relationship with their healthcare professional(s) was an important issue when considering treatment. CONCLUSION: It is concluded that this study has successfully used Q-methodology to systematically investigate people's subjectivity and developed a novel approach to elicit the views of individual patients, as well as explore and differentiate between groups of patients. PRACTICE IMPLICATIONS: The formation of true partnerships between patients and healthcare professionals which will enhance individual patients' ability to self-manage chronic disease. PMID- 15896943 TI - Impact of age, health locus of control and psychological co-morbidity on patients' preferences for shared decision making in general practice. AB - OBJECTIVE: The aim of this study was to explore if patients' preferences to be involved in decision making correlates with reasons for encounter, psychological or demographic characteristics and with the physicians' estimation of the patients' preferred communication styles. METHODS: Two hundred and thirty-four patients in general practice completed a questionnaire, which included the 'Autonomy Preference Index' (API), 'health locus of control' and 'hospital anxiety and depression'. The physicians documented the diagnoses and estimated the patients' preference for an autonomous, paternalistic or shared decision communication style. RESULTS: The preference for involvement varied across the reason for encounter (range 36.6-50.6), but this effect is explained by the patients' age (p<0.01). The participation preferences and the external and internal health locus of control declined with age whereas depression scores rose (p<0.01). The physicians estimation corresponded with patient API scores (p<0.05). CONCLUSION: It must be questioned if the preference for involvement is really a stable characteristic. The physicians were able to estimate the preferred communication style of their patients presumably on the basis of the patients' age and education. PRACTICE IMPLICATIONS: It appears that not every situation in general practice requires SDM, though different communication styles could be useful if adapted to the individual situation. PMID- 15896944 TI - The preliminary effectiveness of migraine lay trainers in a home-based behavioural management training. AB - OBJECTIVES: This pilot study examined the effectiveness and trainer skills of the first migraine lay trainers (MLTs). METHODS: In a stepwise training program eight MLTs participated in a behavioural management training (BMT) aimed at the prevention of migraine attacks by proactive relaxation and trigger management. After successful reduction of their migraine attacks, three MLTs provided BMT under supervision at home to one fellow patient and subsequently to a small group. RESULTS: Migraine frequency was significantly reduced in five out of eight patients trained by MLTs (mean 48%) and medication use decreased substantially in four patients (mean 47%). Qualities of MLTs concerned their motivational assistance, knowledge of premonitory symptoms and exchange of disease specific problems. Pitfalls were that migraine symptoms hampered an active guidance of the sessions and providing tailored feedback was difficult. CONCLUSION: The first MLTs were successful in training fellow patients in behavioural prevention of migraine attacks. PRACTICE IMPLICATIONS: Continuous supervision of MLTs health and trainer skills is recommended but is likely to have implications for cost effectiveness. PMID- 15896945 TI - C-reactive protein is a significant predictor of decrease in fat mass in hemodialysis patients. AB - Malnutrition and inflammation are common in hemodialysis patients, and are usually closely associated. We examined annual body fat mass changes, a possible nutritional parameter, in maintenance hemodialysis patients, and investigated the factors affecting such changes. Body fat mass of 454 hemodialysis patients (61+/ 11 years, 269 males and 185 females) was measured twice by dual energy X-ray absorptiometry (DEXA), with a 12-month interval between measurements. In a total of 65 patients with hemodialysis duration of less than 1 year at the first measurement, fat mass at the second measurement had increased significantly over the course of a year (P<0.0001). In contrast, in a total of 389 patients with hemodialysis duration of more than 1 year at the first measurement, fat mass at the second measurement significantly decreased (P<0.005). In the 389 patients, significant negative correlations were seen between fat mass changes and CRP (r= 0.165, P<0.005). In a multiple regression analysis, CRP was a significant factor (beta=-0.163, P<0.005) affecting fat mass changes, independent of other confounding clinical factors (R(2)=0.127, P<0.001). These results show that body fat mass of long-term hemodialysis patients decreases after an initial increase in the first to second years of hemodialysis. In hemodialysis patients, fat mass change appears to be a parameter indicative of nutritional changes. Chronic inflammation, represented by higher CRP levels, is a significant factor affecting decrease in fat mass, and is related to poorer nutritional status. PMID- 15896946 TI - DNFB activates MAPKs and upregulates CD40 in skin-derived dendritic cells. AB - BACKGROUND: The intracellular mechanisms involved in the activation of DCs during sensitization in allergic contact dermatitis (ACD) are not known. OBJECTIVE: Here, we investigated the effect of a strong sensitizer, 2,4-dinitrofluorobenzene (DNFB) on the activity of MAPKs in a dendritic cell (DC) line generated from fetal mouse skin (FSDC), and the results were correlated with the expression of a costimulatory molecule upregulated upon DC maturation, CD40. METHODS: Phosphorylation of ERK1/2 (pERK1/2) and p38 MAPK (pp38 MAPK), and CD40 protein levels, were determined by Western blot. Cellular localization of pERK1/2 and pp38 MAPK were determined by immunocytochemistry using phospho-specific antibodies. RESULTS: Although with different kinetics, DNFB activated ERK1/2 and p38 MAPK, and induced the translocation of the phosphorylated forms of the kinases to the nucleus. In addition, DNFB upregulated significantly CD40 protein levels in FSDC. However, 2,4-dichloronitrobenzene (DCNB), an inactive analogue of DNFB, did not affect significantly the phosphorylation of MAPKs and CD40 protein levels. SB203580 and SB202190, inhibitors of the p38 MAPK activity, inhibited DNFB-induced CD40 upregulation, although this effect did not reach statistical significance. In contrast, PD 98059 and U0126, inhibitors of mitogen or extracellular signal-regulated kinase (MEK), had no effect on the CD40 upregulation induced by DNFB. CONCLUSIONS: Taken together, these results indicate that the strong sensitizer DNFB activates ERK1/2 and p38 MAPK signaling pathways, and upregulates CD40 protein levels. However, MAPKs do not play a major role in the induction of CD40, one of the phenotypic markers of DC maturation. PMID- 15896947 TI - The miscibility of poly(D,L-lactide-co-glycolide) with amphiphilic molecules and the interaction of their mixtures with DNA at air/water interface. AB - The miscibility of poly(D,L-lactide-co-glycolide) (PLG) with three amphiphilic molecules and the interaction of the PLG/surfactant mixtures with DNA at air/water interface are investigated by pi-A isotherms, Brewster angle microscopy (BAM) and atomic force microscopy (AFM) techniques. The pi-A isotherms of the PLG mixtures with cationic C(12)AzoC(6)PyBr, and C(12)AzoC(6)N(CH(3))(3)Br, are quite different from the pi-A isotherm of pure PLG on water subphase. In contrast to the case, the pi-A isotherm of PLG mixed with nonionic C(12)AzoC(6)OPy is almost identical to the pure PLG except some increasing of molecular area. Similar phenomena are observed on DNA subphase. The in situ BAM and ex situ AFM observations demonstrate that the dispersion of PLG at air/water interface becomes good when it mixes with the two cationic surfactants, whereas quite poor due to the phase separation when it mixes with the nonionic amphiphilic molecule. Based on these results we conclude that the cationic surfactants can affect the conformation change of PLG at air/water interface and figure a well miscibility with polymer whereas the nonionic amphiphilic molecule presents poor miscibility. In addition, the even mixing of the PLG and the cationic surfactants is favorable for the adsorption to DNA more effectively. PMID- 15896948 TI - Solid-state characterization of falicaine hydrochloride and isomorphic dyclonine hydrochloride. Part IV. Crystal polymorphism of local anaesthetic drugs. AB - Two homologous local anaesthetic drugs, falicaine (propipocaine) hydrochloride (1 (4-propoxyphenyl)-3-(1-piperidinyl)-1-propanone hydrochloride, PPCHC) and dyclonine hydrochloride (1-(4-butoxyphenyl)-3-(1-piperidinyl)-1-propanone hydrochloride, DCNHC) were characterized by thermal analysis (hot-stage microscopy, differential scanning calorimetry, thermogravimetry), vibrational spectroscopic methods (FTIR-, FT-Raman-spectroscopy), powder X-ray diffractometry, solid-state-/solution-NMR and water-vapor sorption analysis. The formation and thermodynamic stability of the two different solid phases of both the compounds is described and presented in a flow chart and energy/temperature diagram, respectively. Of the two substances investigated, mod. II degrees is the thermodynamically stable form at room temperature. This form is present in commercial products of PPCHC as well as of DCNHC, and it endothermally transforms to the less stable form mod. I, at about 10K prior to the melting points. The stable mod. II degrees crystallizes from all tested solvents. Mod. I crystallizes from the super cooled melt. According to the heat of transition rule, mod. I is the thermodynamically less stable form below the transition temperature (enantiotropism). The transition temperatures as well as the melting points of the stable forms could be experimentally determined. The sorption isotherms show a distinct higher hygroscopicity for the less stable mod. I of PPCHC and DCNHC. Solid-state NMR spectra were used to obtain both structural- and molecular-level mobility information. PMID- 15896949 TI - Synthetic hydrogel niches that promote hMSC viability. AB - Photopolymerized poly(ethylene glycol) (PEG) hydrogels were used as a base platform for the encapsulation and culture of human mesenchymal stem cells (hMSCs). The base PEG formulation presents an environment completely devoid of cell-matrix interactions. As such, viability of hMSCs in unmodified PEG hydrogels is very low. This formulation was modified to contain pendant phosphate groups to facilitate the sequestering of osteopontin within the gel, as well as pendant cell-adhesive RGD peptide sequences, which are found in osteopontin and other cell adhesion proteins. The survivability of hMSCs was examined with culture time and as a function of the gel chemistry to examine the role of cell-matrix interactions in promoting long-term viability. In the absence of any adhesive ligands, hMSC viability drops to 15% after 1 week in culture. However, by incorporating the RGD sequence or pendant phosphate groups this low viability was rescued to 75% and 97%, respectively. It is believed that the phosphate groups promote mineralization of the hydrogel network, and this mineral phase sequesters cell-secreted osteopontin, resulting in enhanced cell-matrix interactions and improved cell viability. PMID- 15896950 TI - The relation of rhinitis to recurrent cough and wheezing: a longitudinal study. AB - BACKGROUND: Recurrent cough can be a clinical manifestation of rhinitis. However, it remains unclear if the association between rhinitis and recurrent cough among children is independent of asthma. OBJECTIVE: The aim of the present study was to determine, in a large longitudinal cohort, whether rhinitis is significantly associated with recurrent cough alone, wheezing alone, or the combination of both symptoms during childhood. METHODS: We investigated determinants of recurrent cough, with or without wheezing, using longitudinal data from the Tucson Children's Respiratory Study. Among the 1246 subjects originally enrolled, 1024 children completed at least one questionnaire between the ages of 6 and 18 years and were included in the present study. In any survey, wheezing was defined as at least one wheezing episode during the past year and recurrent cough as two or more coughing episodes lasting at least 1 week without a cold during the past year. Generalized estimating equations were used to determine significant risk factors. RESULTS: After adjusting for sex, skin test reactivity and parental asthma, both rhinitis (OR = 2.47 CI = 1.84, 3.30) and sinusitis (OR = 1.54 CI = 1.11, 2.14) were associated with an increased risk of recurrent cough plus wheezing. The OR associated with rhinitis were significantly reduced for subjects reporting only recurrent cough or only wheezing (OR = 1.43, CI = 1.03, 1.99; and OR = 1.30, CI = 1.07, 1.58, respectively). Recurrent cough and wheezing, when examined independently, showed different patterns of risk factors. CONCLUSION: We found rhinitis to be an independent risk factor for both recurrent cough and wheezing during childhood. Different pathways may be involved in the association of rhinitis with recurrent cough and wheezing. PMID- 15896951 TI - Model predictive control as a tool for improving the process operation of MSW combustion plants. AB - In this paper a feasibility study is presented on the application of the advanced control strategy called model predictive control (MPC) as a tool for obtaining improved process operation performance for municipal solid waste (MSW) combustion plants. The paper starts with a discussion of the operational objectives and control of such plants, from which a motivation follows for applying MPC to them. This is followed by a discussion on the basic idea behind this advanced control strategy. After that, an MPC-based combustion control system is proposed aimed at tackling a typical MSW combustion control problem and, using this proposed control system, an assessment is made of the improvement in performance that an MPC-based MSW combustion control system can provide in comparison to conventional MSW combustion control systems. This assessment is based on simulations using an experimentally obtained process and disturbance model of a real-life large-scale MSW combustion plant. PMID- 15896952 TI - Metabolism of the 16-androstene steroids in primary cultured porcine hepatocytes. AB - The hepatic metabolism of the 16-androstene steroids was investigated using isolated porcine hepatocytes. This study demonstrated that the liver is capable of producing both phase I and phase II steroid metabolites from 16-androstene steroid precursors. 16-Androstene metabolites were recovered by solid-phase extraction and identified by gas chromatography-mass spectrometry (GC-MS). When 5alpha-androstenone was provided as a substrate, both 3beta- and 3alpha androstenol were produced as well as a metabolite that showed evidence of hydroxylation. Incubations with the various 16-androstene steroids produced metabolic profiles which suggested that the major role of the liver is phase II conjugation. Sulfoconjugated 16-androstene steroids included androstadienol, 5alpha-androstenone, 3beta-, 3alpha-androstenol, and possibly the hydroxylated metabolite of 5alpha-androstenone. It was determined that hydroxysteroid sulfotransferase (HST) is the likely candidate for the sulfoconjugation of the 16 androstene steroids within the liver. Despite the capacity of the hepatocytes to sulfoconjugate the 16-androstene steroids, the principle metabolites produced from incubations with 5alpha-androstenone, 3beta-, and 3alpha-androstenol were glucuronide conjugates, accounting for approximately 68% of all phase II metabolism. These findings underline the importance of steroid conjugation and suggest that hepatic metabolism of the 16-androstene steroids may influence the levels of 5alpha-androstenone present in the circulation, and thus, capable of accumulating in fat. PMID- 15896953 TI - Brain aromatase expression after experimental stroke: topography and time course. AB - Brain aromatase has been shown to be increased in expression after neurotoxic damage and to exert neuroprotection via generation of local oestrogens. The present study investigates the topography and time course of brain aromatase expression after experimental stroke (middle cerebral artery occlusion (MCAO)). Ovariectomised stroke prone spontaneously hypertensive rats underwent distal MCAO by electrocoagulation. Immunohistochemistry revealed increased brain aromatase expression at 24h and 8 days in the cortical penumbra/peri-infarct zones with no increase evident at 2h or 30 days post-MCAO. Double label studies indicate that some of the increased aromatase expression is associated with astrocytic processes. Thus, this is the first evidence that aromatase protein is increased after MCAO and the location (peri-infarct), time course (within 24h) and cellular localisation (astrocytic) indicate the potential for aromatase to promote the survival of cells in the penumbra after experimental stroke by local synthesis of oestrogens. PMID- 15896954 TI - Characterization and adsorption properties of eggshells and eggshell membrane. AB - The objective of this work was to study the chemical and physical characterization of eggshell and eggshell membrane particles prepared from the hen eggshell waste. Under the characterization measurements investigated, it was found that the pore structures of the two biomaterials belong to a typical Type II, indicating that they should be basically characteristic of nonporous materials or materials with macropores or open voids. Further, the chemical composition of the resulting eggshell particle was strongly associated with the presence of carbonate minerals from the Fourier transform infrared (FTIR) spectra. In contrast to the resulting eggshell membrane particle, the presence of functional groups of amines and amides was observable because of its chemical composition of fibrous proteins. From the isotherm data of methylene blue at 25 degrees C, the Freundlich model yielded a somewhat better fit than the Langmuir model. The adsorption isotherms revealed the eggshell biosorbents could only uptake the basic dye of less than 1.0mg/g in aqueous medium, which was attributed to their poor pore properties. PMID- 15896955 TI - Plant parasitic and vector nematodes associated with Asiatic and Oriental hybrid lilies. AB - A survey for distribution and abundance of plant parasitic nematodes in fields grown to Lilium in Himachal Pradesh, India at four study sites viz. Nagrota (at 810 m a.s.l.), Palampur (at 1270 m a.s.l.), Sunder Nagar (at 1400 m a.s.l.) and Chail (at 2250 m a.s.l.) was carried out. Moderate (101-500/200 ml soil) to high (501-1000/200 ml soil) populations of phytonematodes including the vectors for plant viruses (Aphelenchoides avenae, Criconemoides spp., Hoplolaimus spp., Longidorus spp., Paratylenchus spp., Pratylenchus spp., Rhabditis spp., Trichodorus spp., Tylenchoryhnchus spp., Tylenchulus spp. and Xiphinema diversicaudatum) were recorded. Mean population of nematodes was positively correlated with pH in all the study sites, negatively correlated with electrical conductivity (EC), percent organic matter (OM%), available potassium (K) and positively correlated with percent carbon (C%), available nitrogen (N) and available phosphorus (P) in all but one study site. The highest incidence of virus-vector nematodes viz. X. diversicaudatum, Longidorus spp. and Trichodorus spp. was recorded at Palampur. Only Strawberry latent ringspot nepovirus (SLRSV) was detected in Lilium cvs. Star Gazer Max and Galeili by Enzyme Linked Immunosorbent Assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) and in X. diversicaudatum associated with the cultivars by RT-PCR. Cucumis sativus used as bait plants showed SLRSV symptoms after 15 days of nematode inoculation and further SLRSV was again detected by ELISA and RT-PCR in C. sativus plants confirming the transmission of SLRSV by X. diversicaudatum in Lilium. PMID- 15896956 TI - Synthesis of a small library of diketopiperazines as potential inhibitors of calpain. AB - A small library of 2,5-diketopiperazines based on previously reported calpain inhibitors was synthesized. In addition, a concise total synthesis of the structurally related natural product phevalin (2) was accomplished. Despite literature reports that some of the compounds prepared were calpain inhibitors, none of the library members were found to have significant activity against recombinant human calpain I. PMID- 15896957 TI - Structure-activity relationship of N-methyl-bisindolylmaleimide derivatives as cell death inhibitors. AB - A series of N-methyl-bisindolylmaleimide derivatives was synthesized and evaluated as cell death inhibitors. N-Methyl-2-[1-(3-aminopropyl)-1H-indol-3-yl] 3-(1H-indol-3-yl)maleimide (21) was the most potent inhibitor of H2O2-induced necrotic death of human leukemia HL60 cells among them. PMID- 15896958 TI - Acyclic cyanamide-based inhibitors of cathepsin K. AB - Conversion of the proline-derived cyanamide lead to an acyclic cyanamide capable of forming an additional hydrogen bond with cathepsin K resulted in a large increase in inhibitory activity. An X-ray structure of a co-crystal of a cyanamide with cathepsin K confirmed the enzyme interaction. Furthermore, a representative acyclic cyanamide inhibitor 6r was able to attenuate bone resorption in the rat calvarial model. PMID- 15896959 TI - Synthesis and evaluation of isatin derivatives as effective SARS coronavirus 3CL protease inhibitors. AB - N-Substituted isatin derivatives were prepared from the reaction of isatin and various bromides via two steps. Bioactivity assay results (in vitro tests) demonstrated that some of these compounds are potent and selective inhibitors against SARS coronavirus 3CL protease with IC50 values ranging from 0.95 to 17.50 microM. Additionally, isatin 4o exhibited more potent inhibition for SARS coronavirus protease than for other proteases including papain, chymotrypsin, and trypsin. PMID- 15896960 TI - Nitrone derivatives of trolox as neuroprotective agents. AB - Synthesis of nitrone derivatives of trolox is described. Their biological evaluation was performed in vitro for scavenging different free radicals, inhibiting Fe(2+)-induced lipid peroxidation, and in vivo in a permanent middle cerebral artery occlusion model in mice. New compounds exert pharmacological activities comparable to or better than those of trolox or nitrone-type reference compounds. PMID- 15896961 TI - Clinical research 5: quantitative data collection and analysis. AB - This six-part research series is aimed at clinicians who wish to develop research skills, or who have a particular clinical problem that they think could be addressed through research. The series aims to provide insight into the decisions that researchers make in the course of their work, and to also provide a foundation for decisions that nurses may make in applying the findings of a study to practice in their own Unit or Department. The series emphasises the practical issues encountered when undertaking research in critical care settings; readers are encouraged to source research methodology textbooks for more detailed guidance on specific aspects of the research process. A couple of points: 1. It is artificial to describe research as qualitative or quantitative. Studies often include both dimensions. However, for the purposes of this paper/series, this distinction is drawn for clarity of writing. 2. It is common practice for quantitative studies to refer to study 'subjects' and qualitative studies to refer to study 'participants'. For ease of reading, the latter term will be used throughout this series PMID- 15896962 TI - Balance control in peripheral neuropathy: are patients equally unstable under static and dynamic conditions? AB - The aim of this investigation was to assess the potentially different effects of impaired proprioceptive input in balance control under static and dynamic conditions in neuropathic patients. We recruited 20 normal subjects and 27 patients affected by neuropathies known to affect to a different extent large and medium size afferent fibres. The patients comprised 5 with Charcot-Marie-Tooth disease type 1A (CMT1A), 8 with CMT type 2 (CMT2) and 14 with Diabetes polyneuropathy (Diabetics). Measurement of balance during quiet stance on a stabilometric platform showed that sway area (SA) was larger in the CMT2 and Diabetics than normal subjects or in CMT1A, under both eyes open and closed conditions. The estimated conduction velocity (CV) of the group II afferent fibres was lower in CMT2 and Diabetics than in normal subjects and CMT1A. Across all patients, SA increased as a function of the slowing of group II CV. During a dynamic balance task the head A-P displacement was only slightly increased in the patient groups with respect to normals, despite the increased delay at which the head followed displacement of the feet. The unpredictably good performance of all patient groups under dynamic condition, which was at variance with their imbalance during quiet stance, may indicate that these patients learnt to exploit anticipatory postural strategies. PMID- 15896963 TI - Synthesis of 2-(aminocarbonylmethylthio)-1H-imidazoles as novel Capravirine analogues. AB - Different analogues of Capravirine (AG-1549) or S-1153 were prepared by synthesis of 2-(5-benzyl-4-isopropyl-1-methyl-2,3-dihydro-1H-imidazol-2-ylthio)acetamide (3a-c), ethyl [5-benzyl-1-(ethoxymethyl)-4-ethyl-1H-imidazol-2-ylthio]acetate (10), 2-[5-alkyl-4-substituted 1-(pyridin-4-ylmethyl)-1H-imidazol-2 ylthio]acetamides (12a-f), and 2-[5-benzyl-1-(benzyloxymethyl)-4-isopropyl-1H imidazol-2-ylthio]acetamides (14a-l) from their corresponding amino acids through a sequence of reactions: Dakin-West reaction, hydrolysis, condensation with thiocyanate derivatives, alkylation with 2-iodoacetamide and ethyl chloroacetate, and coupling with 4-pyridylmethyl chloride, ethoxymethyl chloride and benzyloxymethyl chloride. All the synthesized compounds were screened for their activity against HIV-1 (wild type) and some of them (especially Capravirine like structures) were found active. PMID- 15896964 TI - Synthesis and evaluation of 3-aryl piperidine analogs as potent and efficacious dopamine D4 receptor agonists. AB - A series of 3-aryl piperidine analogs with 2-piperidinoalkylamino or 2 piperidinoalkyloxy fused bicyclic rings were prepared and found to be potent and efficacious human dopamine D4 agonists. The synthesis and structure-activity relationship (SAR) studies that led to the identification of these compounds are discussed. PMID- 15896965 TI - Synthesis of 2,4,6-trisubstituted pyrimidines as antimalarial agents. AB - A series of 2,4,6-trisubstituted-pyrimidines were synthesized and evaluated for their in vitro antimalarial activity against Plasmodium falciparum. Of the 18 compounds synthesized, 14 compounds have shown MIC in the range of 0.25-2 microg/mL. These compounds are in vitro severalfold more active than pyrimethamine. PMID- 15896966 TI - Photoreceptor cell types in the retina of the tuatara (Sphenodon punctatus) have cone characteristics. AB - The tuatara Sphenodon punctatus, restricted to a few New Zealand offshore islands and now strictly protected, belongs to the Rhynchocephalia, the smallest order of extant reptiles. Earlier light microscopical studies on the retina of this species described photoreceptors with both rod- and cone-like features and the presence of a fovea. A limited amount of retinal material from S. punctatus has now allowed us to prepare the first-ever electron microscopic observations on the eye of this reptile. We were able to distinguish three types of photoreceptor, all with fine structural features characteristic of cone cells. Large single cones as well as double cones had open discs in their outer segments and straight axons with pedicle-type terminals. An additional cone type, characterized by somewhat more slender inner and outer segments, vitreally-displaced cell bodies and oblique axons, resembled short-wavelength cones known from other sauropsids. No cells with rod characteristics could be confirmed in the samples, although they might occur in retinal regions not available for this study. We conclude that the tuatara has cone-like photoreceptors, which-as in other crepuscular or nocturnal reptiles-have acquired rod-like features. The phenotypic adaptations notwithstanding, the set of photoreceptor types is quite typical of the reptilian eye and in some respects reminiscent of those seen in lizards and turtles. PMID- 15896967 TI - Molybdenum blue: binding to collagen fibres and microcrystal formation. AB - Collagen fibres have been shown by transmission electron microscopy to progressively bind the polyoxomolybdate ring-complex, termed molybdenum blue. Nucleation of cuboidal molybdenum blue microcrystals occurs on the surface of the collagen fibres, leading eventually to extensive coating of the fibres with microcrystals. PMID- 15896968 TI - Nanomanipulator-assisted fabrication and characterization of carbon nanotubes inside scanning electron microscope. AB - We have installed two nanomanipulators, which can travel about 20mm with a minimum increment of 1 nm, for manipulation of nanostructured materials inside field-emission scanning electron microscope (FE-SEM). Both manipulators render motions in x, y, and z directions, providing various manipulation freedoms such as moving, bending, cutting, and biasing. In addition, we have conducted in situ characterization of the electrical breakdown of multi-walled carbon nanotubes (MWCNTs). Our results demonstrate the possibility that MWCNTs can be used as a gas sensor. PMID- 15896969 TI - 0.23 eV energy resolution obtained using a cold field-emission gun and a streak imaging technique. AB - We demonstrate that a high energy resolution of 0.23 eV is possible by using a cold field-emission electron gun (CFEG) without a monochromator. We have used a 300 kV transmission electron microscope (Hitachi, HF-3000) equipped with a CFEG and an energy filter (Gatan, GIF2002). Since energy instability is critical for high energy resolution in electron energy-loss spectroscopy, we have applied a high-speed 'streak imaging' acquisition technique, in which a series of time resolved spectra are acquired as a two-dimensional spectrum. With this technique, we can easily record 1000-20,000 spectra with an exposure time of 0.353 ms per spectrum. Instability of less than 1.4 kHz has been corrected in the time resolved spectra, allowing the inherent performance of the CFEG to be realized. PMID- 15896970 TI - Scanning electron microscopy of foreleg tarsal sense organs of the poultry red mite, Dermanyssus gallinae (DeGeer) (Acari:Dermanyssidae). AB - Scanning electron microscopy (SEM) studies on the poultry red mite, Dermanyssus gallinae (DeGeer), showed that the tip of the foreleg tarsus of both sexes and early instar stages (larvae, protonymph and deutonymph) bears a cluster of 10 blunt peg-type sensillae. Diversity in size, shape and surface features of the wall of these sensillae characterize the sensilla complex (sensory field). Two short setae (1 and 2) with smooth walls lack detectable pores, whereas three medium-sized setae (4, 5 and 8) bear pores in the wall. Four medium or long setae (3, 6, 7 and 9) have longitudinal grooves or ribs in the wall, whilst one seta (10) possesses both pores and grooves in the wall. Based on the morphology, it is suggested that the porous setae could be olfactory receptors, and the grooved setae could be mechanoreceptors. PMID- 15896971 TI - Determination of the activity concentration of a 238 Pu solution by the defined solid angle method utilizing a novel dual diaphragm-detector assembly. AB - The activity concentration of a (238)Pu solution was measured by the determined solid angle method employing a novel dual diaphragm-detector assembly, which has been previously described. Due to the special requirements of the detector, a new type of source holder was developed, which consisted of sandwiching the radioisotope between two organic films called VYNS. It was experimentally demonstrated that the VYNS films do not absorb alpha particles, but reduce their energy by an average of 22 keV.A mean activity concentration for (238)Pu of 359.10+/-0.8 kBq/g was measured. PMID- 15896972 TI - Akt/Bad signaling and motor neuron survival after spinal cord injury. AB - The serine-threonine kinase Akt is a cell survival signaling pathway that inactivates the proapoptotic BCL-2 family protein Bad and promotes cell survival in cerebral ischemia. Involvement of the Akt/Bad signaling pathway after spinal cord injury (SCI) is, however, uncertain. Our results showed that phospho-Akt (serine-473) and phospho-Bad (serine-136) were significantly upregulated at 1 day after SCI. In addition, phospho-Akt and phospho-Bad were colocalized in motor neurons that survived SCI and inhibition of PI3-K reduced expression of phospho Akt and phospho-Bad. Dimerization of Bad with 14-3-3 in the cytosol was increased whereas Bad/Bcl-XL binding in the mitochondria was decreased after SCI. We further found that reduced oxidative stress by SOD1 overexpression in rats enhanced the expression of phospho-Akt, phospho-Bad, Bad/14-3-3 binding and reduced Bad/Bcl-XL binding after SCI, as compared to wild-type rats. We conclude that oxidative stress may play a role in modulating Akt/Bad signaling and subsequent motor neuron survival after SCI. PMID- 15896973 TI - Denervation and repeated L-DOPA induce complex regulatory changes in neurochemical phenotypes of striatal neurons: implication of a dopamine D1 dependent mechanism. AB - Motor complications induced through repeated L-DOPA treatment in patients with Parkinson's disease are thought to be the consequence of molecular adaptations that occur in response to repeated dopamine receptors stimulation. Here, we studied the molecular changes taking place in the denervated striatum of unilaterally 6-OHDA-lesioned rats repeatedly treated with L-DOPA alone or combined to the D1 receptor antagonist SCH23390. We looked at the territorial patterns of expression of neurotensin (NT), dynorphin (DYN), enkephalin (ENK) and Nur77 (also known as NGFI-B) mRNA expression in the striatum and contrasted these with markers of glutamatergic transport and dopaminergic receptor functions. The denervation process induced NT and Nur77 mRNA expression in ENK-positive cells. Subsequent repeated L-DOPA treatment led to a sensitization of L-DOPA-induced rotational response and produced a second surge of NT induction, this time limited to DYN-positive cells and preferentially restricted to the lateral striatum. In this specific territory, the number of Nur77-positive cells was decreased, in response to L-DOPA, when compared to the medial part of the lesioned striatum. L-DOPA treatment increased dopamine D3 receptor and glutamate transporter 1 (GLT1) mRNA expression in the lesioned striatum and that, specifically in an area overlapping one of Nur77 decrease and of NT/DYN induction. The concomitant administration of SCH23390 with repeated L-DOPA treatment blocked the development of behavioral sensitization and the appearance of all L-DOPA-induced molecular reorganizations reported above. Our results showed that repeated L-DOPA treatment produces, in a denervated striatum, a complex pattern of genes regulation in both the direct and the indirect striatal output pathways. This phenomenon is located preferentially in a striatal area receiving converging inputs from the thalamus and sensorimotor cortex and is dependent upon D1 receptor stimulation. PMID- 15896974 TI - Circulating beta-chemokines and matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 system in hemodialyzed patients--role of oxidative stress. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), beta chemokines, increased oxidative stress (SOX) and inflammation have been implicated as important factors in atherosclerosis and vascular remodeling. We hypothesized the possible roles of beta-chemokines [monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory proteins (MIP-1alpha, MIP-1beta) and regulated upon activation, normal T-cell expressed and secreted (RANTES)] as regulators of the metabolism of the vascular extracellular matrix in conditions of increased SOX in hemodialysis (HD) patients. We compared pre-dialysis levels of MMP-9/TIMP-1 system, beta-chemokines, Cu/Zn superoxide dismutase (Cu/Zn SOD) as a marker of SOX and C-reactive protein (CRP) as a marker of inflammation in HD patients with and without cardiovascular disease (CVD) to those of controls. HD patients, particularly those with CVD, showed a significant increase in values of Cu/Zn SOD, CRP, TIMP-1, TIMP-1/MMP-9 ratio, MCP-1 and MIP-1beta, whereas RANTES levels were lower than in the controls. The levels of MIP-1alpha as well as MMP-9 in all HD groups were similar to the controls. The positive correlations were observed between the MMP-9/TIMP-1 system and beta-chemokines, SOX and inflammation in whole HD group and in the subgroup with CVD. Multivariate analysis showed that the duration of dialysis followed by Cu/Zn SOD, MIP-1alpha and beta levels were the significant positive predictors of TIMP-1. In conclusion, our data show that MMP-9/TIMP-1 system and beta-chemokines could cooperate in conditions of elevated SOX, which ultimately predisposes hemodialysis patients to accelerated atherosclerosis. PMID- 15896975 TI - Hepato- and neurotoxicity induced by thioacetamide: protective effects of melatonin and dimethylsulfoxide. AB - The effects of melatonin and dimethylsulfoxide (DMSO) on liver and brain oxidative stress, hepatic failure and blood urea nitrogen (BUN) level changes produced by a single dose of thioacetamide (TAA) in Wistar rats were studies. A dose of either melatonin (3 mg kg(-1)day(-1)) or DMSO (2 g kg(-1)day(-1)) was injected for 3 days before and for 2 days after the administration of TAA (150 mg kg(-1) i.p.). Blood samples were taken from the neck vascular in order to determine ammonium, BUN and liver enzymes. We estimated lipid peroxidation products, reduced glutathione (GSH) content and catalase activity in liver and brain homogenates. TAA caused significant increases in ammonium and in the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) enzymes, while it decreased BUN values. TAA also increased lipid peroxidation product levels, but reduced GSH content and catalase activity in the liver and brain. Both melatonin and DMSO, although melatonin more significantly, decreased the intensity of the changes produced by the administration of TAA alone. Furthermore, melatonin alone or combined with TAA increased the BUN levels and decreased the ammonia values compared with control animals. These results support the antioxidative and neuro-/hepato-protective action of melatonin and a lesser action of DMSO. Likewise, these data seem to support the hypothesis of an effect of melatonin on urea synthesis. PMID- 15896976 TI - Evaluation of antioxidant activity of Brazilian plants. AB - In this work, 22 alcoholic extracts, obtained from 14 species of plants belonging to four families, used for different food and medicinal purposes in Brazil, were evaluated for their capacity to inhibit the reduction of the free radical, 1,1 diphenyl-2-picrylhydrazyl (DPPH), and to protect Saccharomyces cerevisiae cells, an eukaryotic cell model, against the lethal oxidative stress caused by tert butylhydroperoxide (TBH). Five extracts, two from Lamiaceae family (ethanol and butanol extracts from aerial parts of Hyptis fasciculata) and three from Palmae family (Copernicia cerifera leaves and mesocarp of fruits and the endocarp/mesocarp of fruits from Orbignya speciosa) were able to increase the tolerance of S. cerevisiae to TBH and showed to be active as DPPH radical scavengers, thus indicating that these plant extracts could be considered as potential sources of antioxidants. With the exception of ethanol extract of H. fasciculata, the remainder four extracts exhibited a DPPH radical scavenging activity higher than that obtained from Ginkgo biloba, a reference plant with well documented antioxidant activity. Interestingly, the ethanol extract of G. biloba were not effective for yeast cell protection, reinforcing the antioxidant potential of these extracts. PMID- 15896977 TI - Ginkgo biloba extract ameliorates ischemia reperfusion-induced renal injury in rats. AB - There is increasing evidence to suggest that reactive oxygen metabolites (ROMs) play a role in the pathogenesis of ischemia/reperfusion injury (I/R) in the kidney. This study was designed to determine the possible protective effect of Ginkgo biloba extract (EGb) on renal ischemia/reperfusion (I/R) injury. Wistar albino rats were unilaterally nephrectomized, and 15 days later they were subjected to 45 min of renal pedicle occlusion followed by 6 h of reperfusion. Ginkgo biloba extract (EGb) (50 mg kg(-1) day(-1)) or saline was administered twice, 15 min prior to ischemia and immediately before the reperfusion period. At the end of the treatment period, all rats were decapitated. Kidney samples were taken for histological examination or determination of the renal malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and collagen content. Production of reactive oxidants was monitored by chemiluminescence (CL) assay. Creatinine and urea concentrations in blood were measured for the evaluation of renal function. Tumor necrosis factor-alpha (TNF-alpha) and lactate dehydrogenase (LDH) were also assayed in serum samples. Ischemia/reperfusion caused a significant decrease in GSH level, which was accompanied with significant increases in MDA level, MPO activity and collagen content of kidney tissues. Similarly, serum BUN and creatinine levels, as well as LDH and TNF alpha, were elevated in the I/R group as compared to control group. On the other hand, EGb treatment reversed all these biochemical indices, as well as histopathological alterations, which were induced by I/R. The findings imply that ROMs play a causal role in I/R-induced renal injury and EGb exerts renoprotective effects probably by the radical scavenging and antioxidant activities. PMID- 15896978 TI - "Tumor metabolome": a post-modern genomic view on the role of low molecular weight substances in cancerous cell growth:an introduction. PMID- 15896979 TI - Overexpression of myelin-associated glycoprotein after axotomy of the perforant pathway. AB - Myelin-associated glycoprotein (MAG) contributes to the prevention of axonal regeneration in the adult central nervous system (CNS). However, changes in MAG expression following lesions and the involvement of MAG in the failure of cortical connections to regenerate are still poorly understood. Here, we show that MAG expression is differently regulated in the entorhinal cortex (EC) and the hippocampus in response to axotomy of the perforant pathway. In the EC, MAG mRNA is transiently overexpressed by mature oligodendrocytes after lesion. In the hippocampus, MAG overexpression is accompanied by an increase in the number of MAG-expressing cells. Lastly, the participation of MAG in preventing axonal regeneration was tested in vitro, where neuraminidase treatment of axotomized entorhino-hippocampal cultures potentiates axonal regeneration. These results demonstrate that MAG expression is regulated in response to cortical axotomy, and indicate that it may limit axonal regeneration after CNS injury. PMID- 15896980 TI - Regio- and stereospecific analysis of glycerolipids. AB - In recent years researchers have recognized the potential value of comprehensive lipid profiling (lipidomics), which was invented and promoted by lipidologists who recognized the many valuable applications that grew out of the fields of DNA profiling (genomics) and protein profiling (proteonomics). Through lipid class selective intrasource ionization and subsequent analysis of two-dimensional cross peak intensities, the chemical identity and mass composition of individual molecular species of most lipid classes can now be determined in a chloroform extract. There remains, however, the necessity to distinguish the enantiomers and isobaric regioisomers resulting from enzymatic and chemical reactions, which conventional high performance liquid chromatography/mass spectrometry (HPLC/MS) has been slow to accommodate, and tandem MS unable to provide. While reversed phase HPLC can separate regioisomers, normal-phase HPLC can resolve diastereomers, and chiral-phase HPLC can effect dramatic resolution of enantiomers, the full potential of the combined systems has seldom been exploited. The present chapter calls attention to both recent and earlier combinations of these methodologies with mass spectrometry, which allows the HPLC/ESI (electrospray ionization)-MS/MS separation and identification of enantiomeric diacylglycerols, triacylglycerols, and glycerophospholipids as well as their isobaric regioisomers. These developments permit further expansion of lipid profiling (lipidomics) and better understanding of lipid metabolism. PMID- 15896981 TI - Automated 3-D extraction and evaluation of the inner and outer cortical surfaces using a Laplacian map and partial volume effect classification. AB - Accurate reconstruction of the inner and outer cortical surfaces of the human cerebrum is a critical objective for a wide variety of neuroimaging analysis purposes, including visualization, morphometry, and brain mapping. The Anatomic Segmentation using Proximity (ASP) algorithm, previously developed by our group, provides a topology-preserving cortical surface deformation method that has been extensively used for the aforementioned purposes. However, constraints in the algorithm to ensure topology preservation occasionally produce incorrect thickness measurements due to a restriction in the range of allowable distances between the gray and white matter surfaces. This problem is particularly prominent in pediatric brain images with tightly folded gyri. This paper presents a novel method for improving the conventional ASP algorithm by making use of partial volume information through probabilistic classification in order to allow for topology preservation across a less restricted range of cortical thickness values. The new algorithm also corrects the classification of the insular cortex by masking out subcortical tissues. For 70 pediatric brains, validation experiments for the modified algorithm, Constrained Laplacian ASP (CLASP), were performed by three methods: (i) volume matching between surface-masked gray matter (GM) and conventional tissue-classified GM, (ii) surface matching between simulated and CLASP-extracted surfaces, and (iii) repeatability of the surface reconstruction among 16 MRI scans of the same subject. In the volume-based evaluation, the volume enclosed by the CLASP WM and GM surfaces matched the classified GM volume 13% more accurately than using conventional ASP. In the surface-based evaluation, using synthesized thick cortex, the average difference between simulated and extracted surfaces was 4.6 +/- 1.4 mm for conventional ASP and 0.5 +/- 0.4 mm for CLASP. In a repeatability study, CLASP produced a 30% lower RMS error for the GM surface and a 8% lower RMS error for the WM surface compared with ASP. PMID- 15896982 TI - Hemispheric lateralization of voice onset time (VOT) comparison between depth and scalp EEG recordings. AB - Auditory-evoked potential (AEP)s elicited to French-language voiced stop consonant (/ba/) and voiceless stop consonant (/pa/) were studied in non-language impaired epileptic patients and non-epileptic volunteers. First, depth AEPs recorded from the primary auditory cortex during pre-surgical exploration and scalp AEPs recordings using high resolution EEG (HR EEG-64 channels scalp EEG) were compared in the same patients. Both methods indicated that the processing of voiced and voiceless consonants was based on a temporal auditory coding. /Ba/ elicited a first complex (N1) at the onset of voicing and a second component [release component (RC)] time-locked to release. This processing took place specifically in the left primary auditory cortex. Source modeling of the RC showed that a left-greater-than-right amplitude of source probes (SP) both in epileptic patients with left-hemispheric language dominance [established by means of invasive tests (WADA test) and/or clinical data] and right-handed non epileptic subjects. Our data suggest that the processing of VOT is related to hemispheric dominance for language and that scalp-recorded AEPs may represent an effective, non-invasive method to establish hemispheric dominance for language in clinical settings. This procedure could complement existing methods and could help to detect the dissociation between receptive and expressive language sometimes observed in patients with epilepsy. PMID- 15896983 TI - Seizure-related injuries in a group of young people with epilepsy wearing protective helmets: incidence, types and circumstances. AB - PURPOSE: To provide information on the incidence, types and circumstances of injuries sustained in a group of young people with epilepsy using protective helmets. METHODS: Thirty-three residential students (21 M, 12 F, age range 5-21, mean 14.5 years) attending a special epilepsy centre over 1 year were provided with helmets. The types of protective measures, seizure frequency, types of injuries, circumstances and outcome were recorded. RESULTS: Fourteen thousand seven hundred and fifty-one seizures were recorded in the 33 patients, which resulted in 59 injuries. The seizure-related injury risk was 4/1000 seizures. Scalp and facial bruises were the commonest injury (51%). Additional protective measures, such as bed guards and padding of dinner tables and sinks, were used for 57% of these students. Helmets were in use in 46% of the accidents; 68% of these accidents resulted in facial or scalp injuries, which required medical attention in 48%. Helmets were not in use in 41% of accidents; 57% of these accidents resulted in facial or scalp injuries, which required medical attention in 36%. Data on wearing of helmets in the accidents were unavailable in 13%. CONCLUSIONS: Injuries continue to occur despite the use of helmets. Changes to the helmet design and modifications to suit the seizure type may improve the protection offered by helmets. PMID- 15896984 TI - Fibroblast growth factor-18 stimulates chondrogenesis and cartilage repair in a rat model of injury-induced osteoarthritis. AB - OBJECTIVE: Osteoarthritis (OA) is the most common form of arthritis and a primary cause of disability, however, there are no treatments that can slow disease progression or repair damaged joint cartilage. Fibroblast growth factor-18 (FGF18) has been reported to have significant anabolic effects on cartilage. We therefore examined its effects on repair of cartilage damage in a rat meniscal tear model of OA. DESIGN: Surgical damage to the meniscus in rats leads to joint instability and significant damage to the articular cartilage at 3 weeks post surgery. At this time, animals received bi-weekly intra-articular injections of FGF18 for 3 weeks, and the knee joints were then harvested for histologic examination. RESULTS: FGF18-induced dose-dependent increases in cartilage thickness of the tibial plateau, due to new cartilage formation at the articular surface and the joint periphery. The generation of new cartilage resulted in significant reductions in cartilage degeneration scores. The highest dose of FGF18 also induced an increase in chondrophyte size and increased remodeling of the subchondral bone. CONCLUSIONS: The results of this study demonstrate that FGF18 can stimulate repair of damaged cartilage in a setting of rapidly progressive OA in rats. PMID- 15896985 TI - Osteoarthritis development in novel experimental mouse models induced by knee joint instability. AB - OBJECTIVE: Although osteoarthritis (OA) is induced by accumulated mechanical stress to joints, little is known about the underlying molecular mechanism. To apply approaches from mouse genomics, this study created experimental mouse OA models by producing instability in the knee joints. METHODS: The models were of four types: severe, moderate, mild, and medial, depending on the severity and direction of instability imposed by combinations of ligament transection and menisectomy. OA development was evaluated by X-ray and histology by Safranin-O staining, and quantified using our original gradings. Expressions of type II, IX and X collagens and matrix metalloproteinase (MMP)-2, -3, -9 and -13 were further examined by immunohistochemistry and in situ hybridization (ISH). RESULTS: The severe, moderate and mild models exhibited OA development in the posterior tibial cartilage. The severe model showed cartilage destruction at 2 weeks and osteophyte formation at 4-8 weeks after surgery; however, the mild model showed only a partial cartilage destruction at 8 weeks. The grading confirmed that the OA disorders progressed depending on the severity of joint instability. In the medial model, the OA development in the medial tibial cartilage was similar to that in the posterior cartilage of the mild model. Among the collagens and MMPs, type X collagen and MMP-13 were markedly induced and colocalized in the early stage OA cartilage. CONCLUSION: We established four types of mouse models exhibiting various speeds of OA progression. By applying a mouse genomics approach to the models, molecular backgrounds in various stages of OA development can be clarified. PMID- 15896986 TI - Performance of the Norwegian version of AUSCAN--a disease-specific measure of hand osteoarthritis. AB - OBJECTIVE: To examine the performance of the Norwegian version of the AUSCAN Index as a disease-specific health status measure in patients with hand osteoarthritis (OA). METHODS: One hundred and ninety-nine patients with clinical hand OA (mean (SD) age 61.7 (5.7) years, 18 (9%) males) underwent a comprehensive examination including joint status, examination of grip strength and completion of several self-reported health status questionnaires. The Australian/Canadian OA hand index (AUSCAN) captures three different dimensions of hand OA: pain (5 items), stiffness (1 item), and difficulties with daily activities (9 items). Our pre-study hypothesis was to identify AUSCAN as a specific hand measure with strong correlations to hand measures and lower correlations to other general measures of health. RESULTS: Patient completion of the AUSCAN Index was similar or better than other measures. The internal consistency of the AUSCAN was excellent. The pain and physical dimension of AUSCAN correlated substantially to each other and moderately to the stiffness scale. The AUSCAN physical scale correlated moderately to substantially to other measures, the highest correlation being seen with the Arthritis Impact Measurement Scale (AIMS) 2 hand and finger function scale (r=0.73). The standardised differences between patients with and without radiographic abnormalities were numerically larger for the AUSCAN pain and physical scales than for other measures. CONCLUSION: The Norwegian version of the AUSCAN has an acceptable clinimetric performance and is a suitable tool for assessment of hand OA. PMID- 15896987 TI - Control of the hypoxic response through regulation of mRNA translation. AB - Hypoxia is a common feature of most solid tumors which negatively impacts their treatment response. This is due in part to the biological changes that result from a coordinated cellular response to hypoxia. A large part of this response is driven by a transcriptional program initiated via stabilization of HIF, promoting both angiogenesis and cell survival. However, hypoxia also results in a rapid inhibition of protein synthesis which occurs through the repression of the initiation step of mRNA translation. This inhibition is fully reversible and occurs in all cell lines tested to date. Inhibition of translation is mediated by two distinct mechanisms during hypoxia. The first is through phosphorylation and inhibition of an essential eukaryotic initiation factor, eIF2alpha. Phosphorylation of this factor occurs through activation of the PERK kinase as part of a coordinated ER stress response program known as the UPR. Activation of this program promotes cell survival during hypoxia and facilitates tumor growth. Translation during hypoxia can also be inhibited through the inactivation of a second eukaryotic initiation complex, eIF4F. At least part of this inhibition is mediated through a REDD1 and TSC1/TSC2 dependent inhibition of the mTOR kinase. Inhibition of mRNA translation is hypothesized to affect the cellular tolerance to hypoxia in part by promoting energy homeostasis. However, regulation of translation also results in a specific increase in the synthesis of a subset of hypoxia induced proteins. Consequently, both arms of translational control during hypoxia influence hypoxia induced gene expression and the hypoxic phenotype. PMID- 15896988 TI - Taf1: a class II transposon of Aspergillus fumigatus. AB - Aspergillus fumigatus is an important medical pathogen that lacks a known sexual cycle. Transposons may provide an important mechanism for the generation of genetic diversity in this organism. Here, we describe Taf1, the first class II transposon to be identified in A. fumigatus. Taf1, a member of the mariner superfamily and pogo family of transposons, is distinguished by the presence of extremely long (89 bp) inverted repeats that flank the transposase coding sequence. Taf1 is present in different locations and copy number among clinical strains of A. fumigatus and is transcribed. Analysis of multiple insertion sequences within a single strain suggests that Taf1 elements undergo inactivation by a repeat induced polymorphism-like mechanism. Taf1 insertion patterns were extremely stable despite multiple stressors including heat shock, serial passage, and infection in mice. Thus Taf1 may be useful for strain identification and molecular typing. PMID- 15896989 TI - Siderophore accumulation and phytopathogenicity in Microbotryum violaceum. AB - We investigated the role of the siderophore rhodotorulic acid (RA) accumulation in the host-pathogen interaction of Microbotryum violaceum and Silene latifolia. While the wild-type M. violaceum accumulated RA in the culture medium in response to iron stress, the monogenic mutant 45 of M. violaceum did not accumulate detectable amounts of RA under limiting iron supply. Genetic analysis showed the mutant 45 locus to be 3.8 cM from the centromere, endorsing the pericentric gene clustering of M. violaceum. Crosses of mutant and wild-type strains demonstrated that the lack of siderophore accumulation in mutant 45 did not preclude pathogenesis, hence fungal reproduction. PMID- 15896990 TI - Ectopic expression of a constitutively active Cdc42 small GTPase alters the morphology of haploid and dikaryotic hyphae in the filamentous homobasidiomycete Schizophyllum commune. AB - Cloning of the Cdc42 gene from Schizophyllum commune enabled investigation of the role of ScCdc42 in the regulation of vegetative growth and sexual reproduction in this fungus, which has a well-characterized hyphal cell structure, cytoskeleton, and mating system. Ectopic expression of the constitutively active Sccdc42(G12V) or Sccdc42(Q61L) alleles from native or inducible ScCel1 promoters in haploid hyphae had dramatic effects on hyphal morphology, cytoskeletal structure, and Cdc42 localization. For transformants with constitutively active Sccdc42, polar tip growth of apical cells in the leading hyphae was normal but polar tip growth in side branches was altered, implying different regulation of polarity establishment in the two groups of apical cells. Branch emergence at exceptional sites and isotropic growth of cells near the septum indicated that ScCdc42 regulates branch site selection and subsequent hyphal development. Poor dikaryotization along with irregular clamp connections in mates expressing Sccdc42(G12V) or Sccdc42(Q61L) suggested that Cdc42 also contributes to efficient mating in S. commune. PMID- 15896991 TI - Features and functions of covalently linked proteins in fungal cell walls. AB - The cell walls of many ascomycetous yeasts consist of an internal network of stress-bearing polysaccharides, which serve as a scaffold for a dense external layer of glycoproteins. GPI-modified proteins are the most abundant cell wall proteins and often display a common organization. Their C-terminus can link them covalently to the polysaccharide network, they possess an internal serine- and threonine-rich spacer domain, and the N-terminal region contains a functional domain. Other proteins bind to the polysaccharide network through a mild-alkali sensitive linkage. Many cell wall proteins are carbohydrate/glycan-modifying enzymes; adhesion proteins are prominent; proteins involved in iron uptake are present, and also specialized proteins that probably help the fungus to survive in its natural environment. The protein composition of the cell wall depends on environmental conditions and developmental stage. We present evidence that the cell wall of mycelial species of the Ascomycotina is similarly organized and contains glycoproteins with comparable functions. PMID- 15896992 TI - Assessing the reliability of a QSAR model's predictions. AB - Quantitative structure activity relationships (QSAR) are one of the well developed areas in computational chemistry. In this field, many successful predictive models have been developed for various property, activity or toxicity predictions. However, the predictive power of models for new query compounds is often not well characterized. The breadth of applicability of models is often not characterized. In other words, with a given QSAR model and a specific query compound to be predicted, can the model be used reliably for the desired prediction? In this study, we assessed the reliability of QSAR models' prediction on query compounds. Our approach, employing hierarchical clustering, was developed and tested using a test dataset containing 322 organic compounds with fathead minnow acute aquatic toxicity as the activity of interest. The hypothesis of the approach was that if a query compound is more similar to the compounds used to generate the QSAR model, it should be predicted more accurately. Thus, the core of the approach is to determine the relationship between the similarity of query compounds to the training set compounds of the QSAR model and the prediction accuracy given by that model. This relationship determination was achieved by comparing the results given by the two major components of the approach: objects clustering and activity prediction. With the resultant information from the two steps, a direct relationship was shown. PMID- 15896993 TI - A model for the interaction between plant GAPN and 14-3-3zeta using protein protein docking calculations, electrostatic potentials and kinetics. AB - Phosphorylated non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.9; GAPN) found in heterotrophic cells of wheat is activated by MgCl(2). The divalent cation disrupts the interaction between GAPN and a 14-3-3 regulatory protein. This effect is quite remarkable, since it has previously been shown that 14-3-3 binding to a target protein requires divalent cations as Mg(2+) or Ca(2+). Binding of the divalent cation to 14-3-3 causes an increase in surface hydrophobicity. Crystal structure of a 14-3-3-target protein complex has been only determined for serotinin N-acetyltransferase. We utilized a model of a subunit of plant GAPN and the crystallographic structure of human 14-3-3zeta to shape the complex between theses two proteins. Initial dockings were performed with the BiGGER program, which allows an exhaustive search of translational and rotational space. A filtering procedure was then applied to reduce the number of complexes to a manageable number. We predict the structural characteristics of GAPN-14-3-3zeta binding process, proposing that the main attractive force in this complex derives from electrostatic interactions. The predicted model was corroborated by analysis of kinetic behavior of GAPN and its relationship with pH and ionic strength conditions. This study provides a variant on the interaction of 14-3-3 with target proteins, thus affording a wider scenario to establish possible structural models for this remarkable family of regulatory proteins. PMID- 15896994 TI - [The European network of transfusion medicine societies (EuroNet-TMS): The White Book 2005]. AB - Europe is building up. It develops in a quite complex environment, in which health care represents an important field of activities. As for blood transfusion, it plays a major role especially in the development of medical activities as well as for the patients treatments. Today, blood components are still of human origin and there are no substitutes for them. As a medical discipline, Blood Transfusion represents a broad field in medicine which requests the involvement of numerous actors. It is up to professional medical/scientific societies to promote the discipline. This is why it has been considered necessary and relevant to build up a federation of transfusion medicine societies throughout the European Union (EU) ; it is called EuroNet-TMS, the European Network of Transfusion Medicine Societies. This network groups more than 7500 professionals of involved in blood transfusion activities. It has six major objectives: 1) To find coherent responses to issues at stake in transfusion; 2) To promote medical and scientific developments of blood transfusion in Europe; 3) To ensure the highest and most up-to-date scientific level to meet safety and quality standards; 4) To offer similar services to all EU citizens in the field of blood transfusion; 5) To share knowledge and date within Europe; 6) To develop interfaces with decision-makers among the diverse European countries. The first step is the writing of the "White Book 2005" which reports the state of the art of blood transfusion in Europe; a prospective plan is proposed to be discussed. PMID- 15896995 TI - The medical imaging interaction toolkit. AB - Thoroughly designed, open-source toolkits emerge to boost progress in medical imaging. The Insight Toolkit (ITK) provides this for the algorithmic scope of medical imaging, especially for segmentation and registration. But medical imaging algorithms have to be clinically applied to be useful, which additionally requires visualization and interaction. The Visualization Toolkit (VTK) has powerful visualization capabilities, but only low-level support for interaction. In this paper, we present the Medical Imaging Interaction Toolkit (MITK). The goal of MITK is to significantly reduce the effort required to construct specifically tailored, interactive applications for medical image analysis. MITK allows an easy combination of algorithms developed by ITK with visualizations created by VTK and extends these two toolkits with those features, which are outside the scope of both. MITK adds support for complex interactions with multiple states as well as undo-capabilities, a very important prerequisite for convenient user interfaces. Furthermore, MITK facilitates the realization of multiple, different views of the same data (as a multiplanar reconstruction and a 3D rendering) and supports the visualization of 3D+t data, whereas VTK is only designed to create one kind of view of 2D or 3D data. MITK reuses virtually everything from ITK and VTK. Thus, it is not at all a competitor to ITK or VTK, but an extension, which eases the combination of both and adds the features required for interactive, convenient to use medical imaging software. MITK is an open-source project (www.mitk.org). PMID- 15896996 TI - Towards a model of lung biomechanics: pulmonary kinematics via registration of serial lung images. AB - The lungs are highly elastic organs, composed of a variety of structures: vasculature, airways and parenchyma. The unique mechanical properties of each of these structures form the composite material of the lung. Numerous pulmonary diseases affect these material properties. Clinically, these structural changes cannot be directly quantified. However, medical imaging modalities such as computed tomography and magnetic resonance imaging can be used to observe lung morphology. It would be helpful to be able to correlate regional morphological changes with changes in pulmonary function. We present an approach toward the quantification of pulmonary deformation via non-rigid registration of serial MR images of the lung using the variational framework implemented in the Insight toolkit. Conventional registration methods, as exemplified by a finite element implementation of the classic elastic matching technique, are shown to perform well over a set of vascular landmarks in the measurement of lung motion. This performance is maintained in an augmented system, which combines inhomogeneous material properties with the use of domain discretizations tailored to reflect the apparent geometry within the image and to reduce background effects. These adaptations lay the groundwork for biomechanical modeling of the lung using the finite element method. PMID- 15896997 TI - A hybrid framework for 3D medical image segmentation. AB - In this paper we propose a novel hybrid 3D segmentation framework which combines Gibbs models, marching cubes and deformable models. In the framework, first we construct a new Gibbs model whose energy function is defined on a high order clique system. The new model includes both region and boundary information during segmentation. Next we improve the original marching cubes method to construct 3D meshes from Gibbs models' output. The 3D mesh serves as the initial geometry of the deformable model. Then we deform the deformable model using external image forces so that the model converges to the object surface. We run the Gibbs model and the deformable model recursively by updating the Gibbs model's parameters using the region and boundary information in the deformable model segmentation result. In our approach, the hybrid combination of region-based methods and boundary-based methods results in improved segmentations of complex structures. The benefit of the methodology is that it produces high quality segmentations of 3D structures using little prior information and minimal user intervention. The modules in this segmentation methodology are developed within the context of the Insight ToolKit (ITK). We present experimental segmentation results of brain tumors and evaluate our method by comparing experimental results with expert manual segmentations. The evaluation results show that the methodology achieves high quality segmentation results with computational efficiency. We also present segmentation results of other clinical objects to illustrate the strength of the methodology as a generic segmentation framework. PMID- 15896998 TI - Symmetric image registration. AB - This paper presents an original non-rigid image registration approach, which tends to improve the registration by establishing a symmetric image interdependence. In order to gather more information about the image transformation it measures the image similarity in both registration directions. The presented solution is based on the interaction between the images involved in the registration process. Images interact through forces, which according to Newton's action-reaction law form a symmetric relationship. These forces may transform both of the images, although in our implementation one of the images remains fixed. The experiments performed to demonstrate the advantages of the symmetric registration approach involve the registration of simple objects, the recovery of synthetic deformation, and the inter-patient registration of real images of the head. The results show that the symmetric approach improves both the registration consistency and the registration correctness. PMID- 15897000 TI - Efficient multi-modal dense field non-rigid registration: alignment of histological and section images. AB - We describe a new algorithm for non-rigid registration capable of estimating a constrained dense displacement field from multi-modal image data. We applied this algorithm to capture non-rigid deformation between digital images of histological slides and digital flat-bed scanned images of cryotomed sections of the larynx, and carried out validation experiments to measure the effectiveness of the algorithm. The implementation was carried out by extending the open-source Insight ToolKit software. In diagnostic imaging of cancer of the larynx, imaging modalities sensitive to both anatomy (such as MRI and CT) and function (PET) are valuable. However, these modalities differ in their capability to discriminate the margins of tumor. Gold standard tumor margins can be obtained from histological images from cryotomed sections of the larynx. Unfortunately, the process of freezing, fixation, cryotoming and staining the tissue to create histological images introduces non-rigid deformations and significant contrast changes. We demonstrate that the non-rigid registration algorithm we present is able to capture these deformations and the algorithm allows us to align histological images with scanned images of the larynx. Our non-rigid registration algorithm constructs a deformation field to warp one image onto another. The algorithm measures image similarity using a mutual information similarity criterion, and avoids spurious deformations due to noise by constraining the estimated deformation field with a linear elastic regularization term. The finite element method is used to represent the deformation field, and our implementation enables us to assign inhomogeneous material characteristics so that hard regions resist internal deformation whereas soft regions are more pliant. A gradient descent optimization strategy is used and this has enabled rapid and accurate convergence to the desired estimate of the deformation field. A further acceleration in speed without cost of accuracy is achieved by using an adaptive mesh refinement strategy. PMID- 15897001 TI - Environmental genotoxicity assessment of an urban stream using freshwater planarians. AB - Pollution is a major concern in urban areas. Due to its biological significance, genotoxicity should be a main focus for pollution biomonitoring, due mainly to the increasing complexity of the chemical environment in which organisms are exposed. Diluvio's Basin (Porto Alegre, RS, Brazil) is a heavily polluted urban ecosystem impacted by urban wastewater. Planarians are useful organism for evaluating environmental genotoxicity because of their high sensitivity, low cost, high proliferative rate and also because of their basal evolutionary position in relation to complex metazoans. Comet assay is a powerful and highly sensitive method of evaluating primary DNA lesions. Based on the unique features of planarians and the current environmental state of Diluvio's Basin, the aim of this work was to evaluate the genotoxic potential of this body of water using comet assay in planarians. Planarians were exposed to the water for 13 days in a laboratory and comet assay was performed in order to screen possible DNA damages. The results indicated an increasing gradient of damage towards basin's mouth. Such a gradient could be related to the gradual increase of pollutants among the different sample sites. Moreover, there seems to be a correlation between the urbanization gradient that exists within the watershed and the genotoxicity. Historical physical-chemical data was also gathered and examined for possible correlations with genotoxicity. Comet assay in planarians is a very promising test for environmental monitoring studies. Its application should be expanded. PMID- 15897002 TI - Comparative studies of the defense mechanism against Schistosoma japonicum of schistosome-susceptible and -resistant Oncomelania nosophora. AB - The amphibious snail Oncomelania nosophora is an intermediate host of Schistosoma japonicum. Previously we reported that there are two strains of the snail, one resistant and one susceptible to a Mindoro, the Philippines, strain of S. japonicum. The resistant snails were collected from Nirasaki and susceptible snails from Kisarazu, Japan. To determine early cellular responses in the two snail strains, we examined histologic alterations in the snails for up to 18 h after the initial exposure to miracidia. In both strains, the penetrating miracidia were distributed in the foot, mantle, gills, heart, stomach, and kidney, and the mean number of penetrating miracidia was similar in both strains. After penetration, snail hemocytes migrated toward the larvae, and by 12 h after exposure, substantial numbers of penetrated larvae were surrounded and encapsulated by hemocytes. The percentage of larvae encapsulated by hemocytes during 12-18 h after the exposure was significantly higher in the resistant Nirasaki strain (60.9+/-19.8%) than in the susceptible Kisarazu strain (42.3+/ 15.0%). In a few snails of the Nirasaki strain, all the larvae found were encapsulated by hemocytes. The differences in hemocyte responses between the two strains may explain the susceptibility of the snails to schistosome larvae. PMID- 15897003 TI - Spectral and photophysical properties of intramolecular charge transfer fluorescence probe: 4'-dimethylamino-2,5-dihydroxychalcone. AB - The spectral and photophysical properties of a new intramolecular charge transfer (ICT) probe, namely 4'-dimethylamino-2,5-dihydroxychalcone (DMADHC) were studied in different solvents by using steady-state absorption and emission spectroscopy. Whereas the absorption spectrum undergoes minor change with increasing polarity of the solvents, the fluorescence spectrum experiences a distinct bathochromic shift in the band position and the fluorescence quantum yield increases reaching a maximum before decrease with increasing the solvent polarity. The magnitude of change in the dipole moment was calculated based on the Lippert-Mataga equation. These results give the evidence about the intramolecular charge transfer character in the emitting singlet state of this compound. PMID- 15897004 TI - Spectroscopy, NMR and DFT studies on molecular recognition of crown ether bridged chiral heterotrinuclear salen Zn(II) complex. AB - A barium-containing crown ether bridged chiral heterotrinuclear salen Zn(II) complex BaZn2L(ClO4)2, where L is a folded dinuclear chiral (R,R)-salen ligand, has been synthesized and characterized by elemental analysis, 1H NMR, UV-vis, IR, circular dichroism (CD) spectra, and mass spectra. As a folded dinuclear chiral host, its recognition with achiral guests (imidazole derivatives), rigid bidentate guest (1,4-diazobicyclo[2,2,2]octane, DABCO) and chiral guests (amino acid methyl esters) was investigated by means of UV-vis spectrophotometric titration, CD spectra. The association constants of D-amino acid methyl esters are found to be higher than those of their L-enantiomer. The sandwich-type binding of BaZn2L(ClO4)2-DABCO supramolecular assembly was specially studied via 1H NMR titration and 1H ROESY. To understand the recognition on molecular level, density functional theory (DFT) calculations on B3LYP/LanL2DZ were performed on the minimal energy conformations of host, guests, and host-guest complexes. The minimal energy conformations were obtained by molecular mechanics (MM) optimization and molecular dynamics (MD) simulation. The results of single point energy, HOMO energy, and charges transfer were analyzed. The results of theoretical calculations are in good agreement with the experimental data. PMID- 15897005 TI - Effects of information and machine learning algorithms on word sense disambiguation with small datasets. AB - Current approaches to word sense disambiguation use (and often combine) various machine learning techniques. Most refer to characteristics of the ambiguity and its surrounding words and are based on thousands of examples. Unfortunately, developing large training sets is burdensome, and in response to this challenge, we investigate the use of symbolic knowledge for small datasets. A naive Bayes classifier was trained for 15 words with 100 examples for each. Unified Medical Language System (UMLS) semantic types assigned to concepts found in the sentence and relationships between these semantic types form the knowledge base. The most frequent sense of a word served as the baseline. The effect of increasingly accurate symbolic knowledge was evaluated in nine experimental conditions. Performance was measured by accuracy based on 10-fold cross-validation. The best condition used only the semantic types of the words in the sentence. Accuracy was then on average 10% higher than the baseline; however, it varied from 8% deterioration to 29% improvement. To investigate this large variance, we performed several follow-up evaluations, testing additional algorithms (decision tree and neural network), and gold standards (per expert), but the results did not significantly differ. However, we noted a trend that the best disambiguation was found for words that were the least troublesome to the human evaluators. We conclude that neither algorithm nor individual human behavior cause these large differences, but that the structure of the UMLS Metathesaurus (used to represent senses of ambiguous words) contributes to inaccuracies in the gold standard, leading to varied performance of word sense disambiguation techniques. PMID- 15897006 TI - Cortical generators of slow evoked responses elicited by spatial and nonspatial auditory working memory tasks. AB - OBJECTIVE: Slow evoked responses have been extensively studied using electrophysiological and neuroimaging methods, but there is no consensus regarding their generators. We investigated the generators of the P3 and positive slow wave (PSW) in the evoked responses to probes recorded during auditory working memory tasks to find out whether there is dissociation between functional networks involved in the generation of the P3 and PSW and between spatial and nonspatial auditory processing within this time window. METHODS: Whole-head magneto-(MEG) and electroencephalography (EEG); analysis of MEG data using minimum-norm current estimates. RESULTS: The associative temporal, occipito temporal and parietal areas contributed to the generation of the slow evoked responses. The temporal source increased while the occipito-temporal source diminished activity during transition from the P3 to PSW. The occipito-temporal generator of the P3 was activated more during the spatial than nonspatial task, and the left temporal generator of the PSW tended to be more strongly activated during the nonspatial task. CONCLUSIONS: These findings indicate that partially distinct functional networks generate the P3 and PSW and provide evidence for segregation of spatial and nonspatial auditory information processing in associative areas beyond the supratemporal auditory cortex. SIGNIFICANCE: The present results support the dual-stream model for auditory information processing. PMID- 15897007 TI - Normal values for single fiber EMG parameters of frontalis muscle in healthy subjects older than 70 years. AB - OBJECTIVE: Single fiber EMG (SFEMG) is a potent electrophysiological method to evaluate impaired neuromuscular transmission, and allows sensitive diagnosis of neuromuscular transmission abnormalities such as myasthenia gravis. The jitter and fiber density values are different for various muscles and age groups and the reference values increase with age. In this study, we evaluated the reference values of jitter and fiber density of frontalis muscle in healthy subjects older than 70 years. METHODS: We evaluated the jitter and fiber density of frontalis muscle in 32 healthy subjects. Twenty-two of them were between 70 and 79 years old (mean +/- SD, 73.9 +/- 1.7), and 10 of them were older than 80 years (mean +/ SD, 82.2 +/- 1.2). RESULTS: Normal limit of jitter (95% confidence limit) was calculated as 40.4 micros for healthy subjects between 70 and 79 years old and 43.7 micros for healthy subjects older than 80 years and normal limit of fiber density (95% confidence limit) were calculated as 1.90 for subjects between 70 and 79 years old and 2.14 for subjects older than 80 years. CONCLUSIONS: We designated the reference values of jitter and fiber density for frontalis muscle in healthy subjects older than 70 years. Our reference values may have value to diagnose neuromuscular transmission abnormalities in elderly patients. SIGNIFICANCE: SFEMG is sensitive for neuromuscular transmission abnormalities and it is important to know the reference values of frontalis muscle in healthy subjects older than 70 years. PMID- 15897008 TI - An evaluation of automated neonatal seizure detection methods. AB - OBJECTIVE: To evaluate 3 published automated algorithms for detecting seizures in neonatal EEG. METHODS: One-minute, artifact-free EEG segments consisting of either EEG seizure activity or non-seizure EEG activity were extracted from EEG recordings of 13 neonates. Three published neonatal seizure detection algorithms were tested on each EEG recording. In an attempt to obtain improved detection rates, threshold values in each algorithm were manipulated and the actual algorithms were altered. RESULTS: We tested 43 data files containing seizure activity and 34 data files free from seizure activity. The best results for Gotman, Liu and Celka, respectively, were as follows: sensitivities of 62.5, 42.9 and 66.1% along with specificities of 64.0, 90.2 and 56.0%. CONCLUSIONS: The levels of performance achieved by the seizure detection algorithms are not high enough for use in a clinical environment. The algorithm performance figures for our data set are considerably worse than those quoted in the original algorithm source papers. The overlap of frequency characteristics of seizure and non seizure EEG, artifacts and natural variances in the neonatal EEG cause a great problem to the seizure detection algorithms. SIGNIFICANCE: This study shows the difficulties involved in detecting seizures in neonates and the lack of a reliable detection scheme for clinical use. It is clear from this study that while each algorithm does produce some meaningful information, the information would only be usable in a reliable neonatal seizure detection process when accompanied by more complex analysis, and more advanced classifiers. PMID- 15897010 TI - Cytokines and fibrinolytic enzymes in tuberculous and parapneumonic effusions. AB - Tuberculous (TB) pleurisy and parapneumonic effusion (PPE) are common causes of pleural fibrosis. The mechanisms underlying fibrin deposition may be different since involved inflammatory cells are distinct. In this study, we measured various cytokines and fibrinolytic enzymes and compared the differences between the two effusions. PPE was further divided into noncomplicated PPE and complicated PPE/empyema subgroups. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, IL-8, macrophage inflammatory protein (MIP)-1beta, monocyte chemoattractant protein (MCP)-1, plasminogen activator inhibitor type 1 (PAI-1) and tissue type plasminogen activator (tPA) were measured using enzyme-linked immunosorbent assays. Significantly higher values of PAI-1, PAI-1/tPA ratio, IL 1beta, IL-8 and MIP-1beta and significantly lower values of TNF-alpha, IL-6 and MCP-1 were observed in PPE/empyema than in TB effusions. Compared to noncomplicated PPE, complicated PPE/empyema had significantly higher levels of TNF-alpha, IL-1beta, IL-8 and MIP-1beta. TB pleurisy patients who had higher effusion levels of TNF-alpha, IL-1beta and IL-8 were predisposing to residual pleural thickening. The underlying mechanisms of fibrin formation and deposition between the two effusions studied (PPE/empyema and TB pleurisy) could not be fully explained by the results of the present study. More studies are needed to explore this further. PMID- 15897011 TI - Identification of GRASP-1 as a novel 97 kDa autoantigen localized to endosomes. AB - We have identified an autoantigen that is recognized by antibodies from an 18 year-old female with a history of recurrent infections who later in her clinical course developed Raynaud's phenomenon and telangiectasias. By indirect immunofluorescence (IIF), the index serum produced a unique cytoplasmic discrete speckled (CDS) staining pattern that partially colocalized with early endosome antigen 1 (EEA1) but not Golgi complex or other cytoplasmic organelles in HEp-2 cells. When HEp-2 cells were treated with 0.1 N HCl, the cytoplasmic speckled staining of the index serum was markedly decreased, suggesting that the reactive antigen was soluble. Western blot analysis showed a reactive approximately 97 kDa protein in a saline soluble protein preparation from HeLa cells. Mass spectrometric analysis of the excised 97 kDa band that was immunoprecipitated from HeLa cell extracts identified GRASP-1 as a possible target. The index serum and anti-GRASP-1 antibodies colocalized to structures in the cytoplasm of HEp-2 cells. Synthetic peptides representing the full-length GRASP-1 protein were used to identify reactive epitopes. Like many other cytoplasmic autoantigens, GRASP-1 has numerous coiled-coil domains throughout the protein with the exception of short segments at the amino and carboxyl terminus. PMID- 15897012 TI - Cloning and analysis of non-specific cytotoxic cell receptor (NCCRP)-1 from common carp Cyprinus carpio L. AB - Nonspecific cytotoxic cell receptor protein (NCCRP-1) provides an important function in target cell recognition and activation of cytotoxicity. NCCRP-1 has been cloned from common carp Cyprinus carpio L. from fish barbel by EST analysis. The isolated gene is composed of 945 bp with a 79 bp 5' UTR, 714 bp open reading frame and 152 bp 3' UTR. The predicted NCCRP-1 gene is composed of 237 amino acid residues and its predicted signal peptide is 19 amino acid residues in length. This gene has conservation of all the related domains characteristic to the NCCRP 1 gene in fish. Phylogenetic and genomic analyses showed that carp NCCRP-1 was similar to other fish orthologues. The expression of NCCRP-1 gene was constitutive in both lymphoid and non-lymphoid tissues. Furthermore, by semi quantitative RT-PCR studies, we showed that NCCRP-1 gene expression is increased in anterior kidney challenged with Aeromonas hydrophila. PMID- 15897013 TI - Glutamate receptors within the nucleus of solitary tract contribute to pancreatic secretion stimulated by intraduodenal hypertonic saline. AB - It is well known that central transmission of vago-vagal reflex within the nucleus of solitary tract (NST) plays an important role in the regulation of gastrointestinal functions. The present study was designed to assess the role of NST glutamate receptor mechanism in pancreatic secretion evoked by intraduodenal hypertonic saline (HS) in anesthetized rats. Intraduodenal infusion of HS significantly (P<0.01) stimulated pancreatic protein output (from 2.60+/-0.09 to 4.18+/-0.24 mg/15 min). Bilaterally microinjected L-glutamate (5 nmol) into the medial nucleus of solitary tract (mNST) produced a significant increase of pancreatic protein secretion (from 2.65+/-0.12 to 4.80+/-0.34 mg/15 min, P<0.01). Bilateral injection of glutamate receptor antagonist kynurenic acid (KYN, 5 nmol) into the mNST completely abolished the increase of pancreatic protein output stimulated by intraduodenal HS (from 4.28+/-0.21 to 2.83+/-0.19 mg/15 min). Either NMDA receptor antagonist dl-2-amino-5-phosphonopentanoic acid (AP5, 1.5 nmol) or AMPA/Kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 1.5 nmol) injected into the mNST markedly attenuated (P<0.05) the pancreatic protein secretion stimulated by intraduodenal HS. In conclusion, these findings showed that blockade of the NST glutamate receptors, including NMDA and AMPA/Kainate receptors antagonized pancreatic secretion evoked by intraduodenal osmolality factor, and suggested that glutamate receptor mechanism within the NST contributed to the central regulation of pancreatic secretion. PMID- 15897014 TI - Identification and functional analysis of TopBP1 and its homologs. AB - The multiple BRCT-domain protein TopBP1 and its yeast homologs have been implicated in many aspects of DNA metabolism, but their molecular functions remain elusive. In this review, we first summarise how the yeast homologs were identified and characterised. We next review the data available from metazoan systems and finally draw parallels with the yeast models. TopBP1 plays important functions in the initiation of DNA replication in all organisms and participates in checkpoint responses both within S phase and following DNA damage. In metazoan systems there is accumulating evidence for additional roles in transcriptional regulation that have not been reported in yeast. Overall, TopBP1 appears to play a key role in integrating different aspects of DNA metabolism, but the mechanistic basis for this remains to be fully explained. PMID- 15897015 TI - Sensitivity enhancement in liquid chromatography/atmospheric pressure ionization mass spectrometry using derivatization and mobile phase additives. AB - High performance liquid chromatography with atmospheric pressure ionization (API) mass spectrometry has been essential to a large number of quantitative analytical applications for a variety of compounds. Poor detection sensitivity however is a problem observed for a number of analytes because detection sensitivity can be affected by many factors. The two most critical factors are the chemical and physical properties of the analyte and the composition of the mobile phase. In order to address these critical factors which may lead to poor sensitivity, either the structure of the analyte must be modified or the mobile phase composition optimized. The introduction of permanently charged moieties or readily ionized species may dramatically improve the ionization efficiency for electrospray ionization (ESI), and thus the sensitivity of detection. Detection sensitivity may also be enhanced via introduction of moieties with high proton affinity or electron affinity. Mobile phase component modification is an alternative way to enhance sensitivity by changing the form of the analytes in solution thereby improving ionization efficiency. pH adjustment and adduct formation have been commonly used to optimize detection conditions. The sensitivity of detection for analytes in bio-matrices could also be enhanced by decreasing ion-suppression from the matrix through derivatization or mobile phase addition. In this review, we will discuss detection-oriented derivatization as well as the application of mobile phase additives to enhance the sensitivity of detection in liquid chromatograph/atmospheric ionization/mass spectrometry (LC/API/MS), focusing in particular on the applications involving small molecules in bio-matrices. PMID- 15897016 TI - High performance liquid chromatographic methods for the determination of aripiprazole with ultraviolet detection in rat plasma and brain: application to the pharmacokinetic study. AB - High performance liquid chromatographic (HPLC) methods were validated for the determination of aripiprazole (OPC-14597, Abilify) in rat plasma and brain. Separation was by Nova-pak phenyl column; flow rate, 1.0 ml/min; mobile phase, acetonitrile-methanol-20 mM sodium sulfate-acetic acid (27:25:48:1, v/v/v/v); UV detection at 254 nm. Reproducibility in plasma and brain showed excellent precision (within 7.8 and 10.6%) and accuracy (96.0-102.4% and 99.0-108.7%) with calibration curve ranges 10.0-2000 ng/ml and 30.0-6000 ng/g, respectively. Validated HPLC methods were successfully applied to pharmacokinetic study of aripiprazole in rats, demonstrating brain concentrations after oral administration five times higher than plasma concentrations. PMID- 15897017 TI - Interference free and simplyfied liquid chromatography-based determination of thiopurine S-methyltransferase activity in erythrocytes. AB - The determination of the thiopurine S-methyltransferase activity (TPMT; EC 2.1.1.67) has become an important issue during thiopurine therapy due to its known genetic polymorphism resulting in a wide range of TPMT activity. Therefore, the standard thiopurine drug regimen is associated with increased hematopoetic toxicity in patients with low or absent TPMT activity, whereas patients with high activity may be insufficiently treated. However, presently available methods are labour intensive and time consuming and tend towards too high or too low enzyme activity due to their methodological approach. The use of instable substrate solutions (6-MP or 6-TG), organic solvents like dimethyl sulfoxide and too high substrate and co-substrate saturation concentrations contribute to this phenomenon. We therefore, established an optimized and fast isocratic HPLC linked TPMT assay based on the enzymatic methylation of mercaptopurine or thioguanine in RBC lysates with S-adenosyl-l-methionine as methyl donor. Unspecific non enzymatic methylation was not detectable. The recovery of 6-methyl-mercaptopurine was 97-102%, the intra- and interday variation between 1.0 and 5.0%, respectively. The assay dispenses with a time consuming extraction procedure with organic solvents, a heating step, and a gradient elution and is therefore, favourable for clinical routine application. The TPMT activity was measured in 62 untreated children with acute lymphoblastic leucemia at the time of diagnosis (activity = 34.0+/-10.6 nmol/g Hb/h, range: 11.5-55.4 nmol/g Hb/h) and in 12 adult healthy volunteers (62.8+/-7.7 nmol/g Hb/h, range: 48-82 nmol/g Hb/h) reflecting the wide measurable TPMT activity found in erythrocytes. PMID- 15897018 TI - HPLC and tandem detection to monitor conformational properties of biopharmaceuticals. AB - High-performance liquid chromatography (HPLC) with UV, circular dichroism (CD) and intrinsic fluorescence detection was applied to monitor conformational properties of recombinant human interferon alpha2b when performing size exclusion chromatography (SEC) and reversed-phase HPLC (RP-HPLC). In this way native conditions during SEC and structural changes of the protein during RP-HPLC were demonstrated. These results were confirmed by stand-alone fluorescence and CD measurements. With respect to HPLC tandem detection, the fluorescence detector compared favourably to the UV and CD detector regarding linearity, sensitivity and selectivity. SEC combined with intrinsic fluorescence scanning detection permits conformational analysis of small amounts of aggregates in the presence of excess native monomeric protein. In conclusion, HPLC with on-line UV and intrinsic fluorescence detection provides a promising concept for analysing the amount and conformational properties of a biopharmaceutical and its impurities. PMID- 15897019 TI - Determination of 13-O-demethyl tacrolimus in human liver microsomal incubates using liquid chromatography-mass spectrometric assay (LC-MS). AB - A simple, sensitive and specific liquid chromatography coupled electrospray ionization mass spectrometric (LC/ESI/MS) method for the determination of 13-O demethylated metabolite (MI), one of the major metabolites of tacrolimus has been developed. The assay uses 32-demethoxyrapamycin (IS) as the internal standard; ethyl acetate as extraction solvent; a Hypersil-Keystone Beta Basic-18 reversed phase column; and a gradient mobile phase of consisting 0.1% formic acid in water and methanol-acetonitrile (3:49, v/v). Mass detection is performed on a single quadrupole mass spectrometer equipped with an electrospray ionization (ESI) interface and operated in a positive ionization mode. MI in the microsomal incubates was quantitated by computing the peak area ratio (MI/IS) analyzed in single ion monitoring (SIM) mode (m/z: 804 and m/z: 901 for MI and IS, respectively). Precision of the assay was determined by calculating the intra-run and inter-run variation at three concentrations (15, 25, 80 ng/ml); the intra run relative standard deviation (R.S.D.) was less than 10% and ranged from 5.0 to 8.3%; and the inter-run R.S.D. was less than 10% and ranged from 4.6 to 9.6%. The limits of detection was 2 ng/ml. This assay has been used to evaluate the effect of three human immunodeficiency virus (HIV) protease inhibitors on the metabolism of tacrolimus in human liver microsomes. PMID- 15897020 TI - Regulation of human extravillous trophoblast function by membrane-bound peptidases. AB - During human placentation, the invasion of extravillous trophoblasts (EVTs) into maternal decidual tissues, especially toward maternal spiral arteries, is considered an essential process for subsequent normal fetal development. However, the precise regulatory mechanisms to induce EVT invasion toward arteries and/or to protect EVTs from further invasion have not been well understood. Recently, we found that two cell surface peptidases, dipeptidyl peptidase IV (DPPIV) and carboxypeptidase-M (CP-M,) are differentially expressed on EVTs. DPPIV expression was mainly observed on EVTs that had already ceased invasion. CP-M was detected on migrating EVTs including endovascular trophoblasts in the maternal arteries. The enzymatic inhibition of these peptidases affected the invasive property of choriocarcinoma-derived cell lines, BeWo and JEG3 cells. In addition, a chemokine, RANTES, that is one of the substrates for DPPIV, enhanced invasion of EVTs isolated from primary villous explant culture and its receptor, CCR1, was specifically expressed on migrating EVTs toward maternal arteries. Furthermore, a novel membrane-bound cell surface peptidase, named laeverin, was found to be specifically expressed on EVTs that had almost ceased invasion. These findings suggest that membrane-bound peptidases are important factors regulating EVT invasion during early placentation in humans. PMID- 15897021 TI - Effects of damage morphology on cortical bone fragility. AB - Despite a general understanding that bone microdamage has distinct strain dependent morphologies, very little information exists on how different damage morphologies develop and participate in bone fracture. In this study, cortical bone beams were subjected to the primary or tertiary phases of bending fatigue followed by either post-hoc fracture toughness tests or microdamage analysis to determine the sequence in which linear microcracks and diffuse damage form during bending fatigue and how they affect the propensity of bone to fracture. The results demonstrate that, following the primary phase, linear microcracks and diffuse damage are formed on the compressive and tensile sides, respectively (p<0.05). Furthermore, this mode of damage formation results in a greater toughness loss if a fracture crack initiates from the tensile side rather than the compressive side (p<0.05). Continued loading of bone specimens to the tertiary phase, however, leads to further accumulation of damage only on the compressive side (p<0.05), and this mode of damage formation results in a further toughness loss if a fracture crack initiates from the compressive side rather than the tensile side (p<0.05). Thus, cortical bone compartmentalizes the damage morphologies in different regions and the sequence of damage production in different phases of cyclic loading to dissipate energy and resist a catastrophic fracture. PMID- 15897022 TI - Vascularised free fibular flap in bone resection and reconstruction. AB - This paper compares allograft alone and in combination with vascularised free fibular flaps (FFF) to reconstruct long bone defects after tumour excision. We present 33 cases, 21 of these patients had reconstruction with an allograft alone as the initial procedure. Nine patients underwent reconstruction with FFF plus allograft plus iliac crest bone graft (ICG), two patients underwent reconstruction with a FFF and ICG and one patient underwent reconstruction with an allograft, a pedicled fibular flap and a FFF. The allograft was obtained from the Queensland Bone Bank and had been irradiated to 25 000Gy. In our experience (N=21) the complication rates with allograft alone were: delayed union 3, nonunion 7, fractured allograft 6, infection requiring resection of the allograft 3, other infections 2. The revision rate was 48% (10 cases of which five required a free fibular flap) and an average of 1.8 revision procedures were required. In the lower limb cases, the mean time to full weightbearing was 20 months and 40% were full weightbearing at 18 months. We felt that the high complication rate compared with other series may have been related to the irradiation of the graft. FFFs were used in 18 cases, 12 cases were primary reconstructions and six were revision reconstructions. The mean fibular length was 19.4 cm (range 10-29 cm). There were no flap losses and the FFF united at both ends of 11 of 12 primary reconstruction cases. One case had nonunion at one end, giving a union rate of 96% (23 of 24 junctions). When a FFF was used in combination with an allograft as a primary reconstruction, the allograft nonunion rate was 50% (five of 10 cases). The mean time to full weightbearing in the lower limb cases was 7.5 months and 100% were full weightbearing at 18 months. The FFF hastens time to full weightbearing but does not appear to affect the complication rates of allograft. The number of revision procedures required is reduced in the presence of a FFF and is the latter is a useful technique for the salvage of refractory cases. PMID- 15897023 TI - New nomenclature concept of perforator flap. AB - Confusion regarding the perforator flap concept has arisen partly from the use of ill-defined nomenclature without consistency; flaps have been named according to either the proximal vessel, the location harvested or the muscle dissected. Since, a variety of conflicting terms can distort the exact understanding of the flap and the true perforator concept, a precise and scientific system of nomenclature is promptly needed. In order to remedy such confusion, the author reviewed 54 recently published articles and 38 abstracts for vague or inaccurate nomenclatures, and compared the perforator flaps with the conventional flaps. A new nomenclature was then drawn up according to three perforator types: direct cutaneous, septocutaneous and musculocutaneous perforator. Even though only musculocutaneous perforators were considered to be true perforators in the initial concept, some perforator flaps have subsequently been added based on septocutaneous or direct cutaneous perforators. Discrimination is necessary and made possible by use of the following nomenclature: a perforator flap based on a musculocutaneous perforator is named according to the name of the muscle perforated, and perforator flaps based on other types of perforators, are named according to the name of the proximal vessel. The term 'perforator based' further defines those flaps harvested without sacrificing the proximal vessel. This new nomenclature concept would be a great help in discriminating among the various patterns of perforator flaps and also in preventing confusion arising from the misnaming of new perforator flaps in the future. Furthermore, the perforator pattern used in the flap can easily be comprehended, and especially in extremities, various perforator flaps, based on the musculocutaneous or septocutaneous perforator, can plainly be distinguished with this new nomenclature concept. PMID- 15897024 TI - Thoracodorsal artery perforator (TAP) type I V-Y advancement flap in axillary hidradenitis suppurativa. AB - Hidradenitis suppurativa is a chronic debilitating disease. Surgical removal of all apocrine glands in the affected region is the definitive treatment. The resulting wound may either be left to heal secondarily or closed primarily. Secondary healing in the axilla may cause contractures and stiffening of the shoulder. Primary healing requires direct closure, split-skin grafting or local flap application. Direct closure is associated with a high incidence of recurrence compared to skin grafting or flaps. Local flap cover is the ideal method of wound closure after excision of the glands. We have used a thoracodorsal artery perforator (TAP) V-Y advancement flap (type I) to achieve closure as a single-stage procedure successfully in four cases. It is a single stage procedure capable of closing large axillary defects whilst preserving the axillary contour. PMID- 15897025 TI - Anatomy of the proximal cutaneous perforator vessels of the gracilis muscle. AB - An anatomic study was performed to analyse the proximal perforator vessels of the gracilis musculocutaneous flap. Twenty-three cadaver legs preserved by the method of Thiel were carefully dissected 24h after the proximal vascular pedicle was injected with a red silicone mass. Nine additional cadaver legs were injected with ink, to visualise the skin area supplied by the proximal perforators, respectively, clarified by a modified Spalteholz technique to demonstrate the anatomic course of the perforators. A considerable variation in numbers and localisation of proximal cutaneous perforators was found. One to four perforators were seen within an area of 6 x 6 cm(2) at the entrance of the main pedicle into the proximal gracilis muscle. Their external diameter ranged from 0.5 to 1.0 mm. The ink-injections showed an oval shaped angiosome with a mean surface of 88 cm(2) at the level of the proximal gracilis pedicle. We conclude from this anatomic study, that a cutaneous flap based on the medial circumflex femoral gracilis perforators can be harvested by experienced hands bearing in mind the unpredictable perforator-anatomy. PMID- 15897026 TI - The use of the Siamese combined free flap to reconstruct challenging defects: twin and triplet variants. AB - The reconstruction of large and intricate defects may need the use of combined flaps due to either the size or requirement for multiple surfaces. The combination may be between free and pedicled tissue transfer, and combined or connected free flaps classified by Koshima. We will discuss the use of the Siamese combined free flap as a method of the reconstructing challenging cases, including one of the largest free tissue transfer reported. PMID- 15897027 TI - A new free flap model in the rat: the pectoralis major muscle. AB - We have designed a rat pectoralis muscle free flap transplant at the level of the pubis. The first choice of arterial pedicle was the thoraco-acromial artery at the level of its branching from the axillary artery. The venous pedicle was the thoraco-acrominal vein. This first technique was attempted on six rats with several failures due to the small diameter of the axillary artery. For this reason, we were obliged to use the axillary vessels. This new technique was performed in 10 rats with positive results in nine cases. The new transplantation of the pectoralis major muscle flap with nerve may be attempted. PMID- 15897028 TI - Covering small defects on the weight bearing surfaces of the foot: the free temporal fasciocutaneous flap. AB - Although defects in the weight bearing area of the heel can be covered by local flaps, radiodermatitis is a contraindication to these flaps. Thin free flaps, as grafted fascial or muscles flaps and thin fasciocutaneous flaps, are usually the option of choice in these particular defects. These reconstructions are prone to shearing strains resulting in ulceration, hypertrophic scars and hyperkeratosis. The authors present a retrospective study of the reconstruction of six small heel defects with the fasciocutaneous temporal free flap performed between 1996 and 2001. The mean size of the defect was 20 cm(2). All arterial anastomoses were performed end to side on the posterior tibial artery. Despite the flap thinness, swelling was present during 12-25 months and one debulking had to be performed. With a mean follow-up of 32 months, all flaps regained protective sensibility after 7 months. No sliding of the flaps could be noted but there was one transient hyperkeratosis. Although the amount of hair on the transferred flaps decreased spontaneously with time, laser hair removal was performed in two patients for psychological reasons. In conclusion, it seems that in selected cases where local flaps are contraindicated, the fasciocutaneous temporal free flap can offer an excellent alternative for heel reconstruction. Due to its particular architecture, it resembles the complex tissue of the sole of the foot resulting in fewer complications and maintenance of flap durability. PMID- 15897029 TI - Invasion and metastasis markers in cancers. AB - Over 90% of all adults human cancers are of epithelial origin comprising mainly of skin and aero-digestive tract cancers. A significant proportion of our discipline's workload consists of management of these cancers. This review article is to provide clinicians with a summary of the current research findings in invasion and metastasis of epithelial cancers and the translation of some of this information to clinical use particularly related to skin and head and neck cancers (HNSCC). Metastasis is the leading cause of death in cancer patients. Although surgical resection of isolated metastases is beneficial for some patients, the overall efficacy of surgery, chemotherapy or radiotherapy is limited. Clearly, with today's advances in surgery a majority of these primary cancers are resectable and a cure attainable if surgeons could control or inhibit metastasis. PMID- 15897030 TI - The change in the appearance of the vermilion border region caused by the difference of the design in symmetric bilateral incomplete cleft lip repair. AB - In the widely performed methods for repair of bilateral cleft lips, such as Mulliken and Trott methods, the formation of the median tubercle is done with the mucocutaneous flaps from the lateral lips. By this manoeuver, a visible horizontal scar just above the white skin roll of the philtral region tends to result, especially among Asian patients. To avoid the formation of this visible scar, we changed our design in such a way that mucosal flaps, instead of mucocutaneous flaps, are elevated from the lateral lips, and the prolabial white skin roll is preserved to become the final philtral white skin roll. By this change in design, the horizontal scar is shifted to the inferior edge of the vermilion border. In the past 8 years, we have performed this refined method along with the conventional one. In the present study, the five cases of symmetric bilateral incomplete cleft lip from each of the two groups are evaluated: one group with the mucocutaneous flaps and the other with the mucosal flaps. From the review of the cases, it was noted that when the philtral region is formed with the mucocutaneous flaps, the horizontal scar tends to be visible. On the other hand, when it is formed with the mucosal flaps, the scar is less conspicuous, although the white skin roll sometimes becomes less-defined. The refined method involving mucosal flaps from the lateral lips produces a better accepted appearance concerning the prolabial horizontal scar in the symmetric bilateral incomplete cleft lip repair. PMID- 15897031 TI - Hypospadias repair: Byar's two stage operation revisited. AB - BACKGROUND: Hypospadias is a congenital deformity characterised by an abnormally located urethral opening, that could occur anywhere proximal to its normal location on the ventral surface of glans penis to the perineum. Many operations had been described for the management of this deformity. METHODS: One hundred and fifteen patients with hypospadias were treated at the Department of Plastic Surgery, Hospital Kuala Lumpur, Malaysia between September 1987 and December 2002, of which 100 had Byar's procedure performed on them. RESULTS: The age of the patients ranged from neonates to 26 years old. Sixty-seven patients had penoscrotal (58%), 20 had proximal penile (18%), 13 had distal penile (11%) and 15 had subcoronal hypospadias (13%). Operations performed were Byar's two-staged (100), Bracka's two-staged (11), flip-flap (2) and MAGPI operation (2). The most common complication encountered following hypospadias surgery was urethral fistula at a rate of 18%. CONCLUSIONS: There is a higher incidence of proximal hypospadias in the Malaysian community. Byar's procedure is a very versatile technique and can be used for all types of hypospadias. Fistula rate is 18% in this series. PMID- 15897032 TI - Penis reconstruction using three different operative methods. AB - To identify the relative success rates, including aesthetic success, of three penis reconstruction techniques, we reviewed 44 cases of penis reconstruction carried out over the past 12 years. The three operative methods we surveyed involved: lower abdominal pedicled fascia flaps; paraumbilical island flaps; and free forearm flaps. Reconstructions survived in only half of the patients receiving lower abdominal pedicled fascia flaps, but 100% success rates were obtained with paraumbilical island flaps and free forearm flaps. The paraumbilical island flap is safe in terms of its blood supply, and the operative procedure is relatively simple in that it does not require microsurgery. Although, the flap is thicker than the forearm skin flap, its shape is satisfactory in slim patients; furthermore, it can be defatted secondarily. The free forearm skin flap provides the best shape, but skilled microsurgery is necessary to carry out the procedure, and damage is likely to the forearm. In conclusion, the best methods to repair defects in the penis in our experience are the paraumbilical island flap and free forearm flap. Lower abdominal pedicled fascia flaps are unsuitable for penile reconstruction and should be used less often. PMID- 15897033 TI - Comparing the cost of delayed and immediate autologous breast reconstruction in Belgium. AB - This study documents the cost of immediate and delayed DIEP flap breast reconstruction. Immediate reconstruction is more attractive from an economic perspective since it only requires one operation, one anaesthetic procedure and one recovery period in hospital. From the perspective of healthcare budget management, assessing the possible cost savings from immediate reconstruction yields interesting results. Since charges do not reflect the real costs of providing care, we calculated resource costs using the micro-costing method. About 95% of the initial mastectomy costs could be saved when performing an immediate breast reconstruction. This was about 35% of total standard direct and indirect costs due to mastectomy and delayed breast reconstruction. In a growing cost conscious environment of managed care, the economic evaluation should, therefore, encourage the trend towards more immediate reconstructions. PMID- 15897034 TI - Breast expander ports appear to be safe after multiple perforations. AB - It has been suggested in the literature that inflation ports of breast tissue expanders can be a major source of leakage. An experimental study with a 23G and subsequently with a 21G injection needle into the port membrane of three inflation ports was performed. After 100 perforations with each needle per port no leakage was detected and no material from the membrane could be identified in the needle lumens. The authors conclude that multiple needle perforations into the injection port are unlikely to be the cause for deflation of breast tissue expanders due to leakage. PMID- 15897035 TI - Umbilicus reconstruction with modified 'unfolded cylinder' technique. AB - Many techniques have previously been reported for reconstruction of the umbilicus. The common goal of reconstruction is to obtain a stable and aesthetically pleasing umbilicus with sufficient depth. Presented here is the reverse application of an 'unfolded cylinder' design, which has previously been reported for nipple reconstruction. The procedure can be applied with local anaesthesia on an outpatient basis and the drawing of the design is simple. Since the walls of the neoumbilicus are consisted of flaps, which are brought together without any twisting or tension, long-term use of a stent is unnecessary and the result is mostly stable and lasting. Moreover, the depth of the neoumbilicus is adjustable by widening the top rectangle in its design, which can increase it. Therefore, the procedure is suitable for thin as far patients. PMID- 15897036 TI - Intravenous immunoglobulins and plasmapheresis combined treatment in patients with severe toxic epidermal necrolysis: preliminary report. AB - Toxic epidermal necrolysis (TEN) is an acute drug-induced life-threatening disorder characterised by extensive epidermal exfoliation and high rate of mortality. Between October 2000 and April 2003, five severe TEN patients were evaluated using a specific TEN severity-of-illness scale (SCORTEN) and treated for the first time, with a combined therapy using Intravenous Human Immunoglobulins (IVIG) and plasmapheresis. The standardised mortality ratio (SMR) analysis ([Sigma observed deaths/Sigma expected deaths]x100) was applied to establish how IVIG and plasmapheresis treatment could reduce TEN patient mortality. The observed mortality was one out of five patients corresponding to 20%. The expected mortality based on SCORTEN was 3.319 corresponding to 66%. The SMR analysis revealed a 70% reduction in mortality (SMR=0.30; 95% confidence interval, 0.0-0.96). Our series show a low mortality rate (20%) related to the severity of the patients (66% expected mortality). The use of IVIG in association with plasmapheresis has a rational basis and may be effective in severe TEN patients. PMID- 15897037 TI - Gene transfer of bFGF to recipient bed improves survival of ischemic skin flap. AB - BACKGROUND: The recipient bed is a promising target of angiogenic therapy to treat ischemic skin flaps. We delivered basic fibroblast growth factor (bFGF) gene to the recipient bed by a plasmid-based method with electroporation, and assessed the effects on flap viability in a rat dorsal skin flap model. METHODS: A 25 x 90 mm(2) axial skin flap was elevated on the back of male Sprague-Dawley rats. Two days before flap elevation, an expression plasmid vector containing the bFGF gene with the signal sequence was injected into the dorsal muscles beneath the skin flap, and then electroporation was delivered (FGF-E(+) group). As control, rats were injected with a plasmid vector containing LacZ gene (LacZ-E(+) group), instead of bFGF gene. Other groups of animals received plasmid vector containing bFGF (FGF-E(-) group) or LacZ (LacZ-E(-) group) gene without electroporation. Seven days later, the area of necrosis and neovascularisation of the skin flap were evaluated. RESULTS: The bFGF gene was successfully transferred to the dorsal muscles, and bFGF was expressed in muscle tissue. The area of flap necrosis (%) in the FGF-E(+) group (21.7+/-5.3%) was significantly smaller than that in the LacZ-E(+) (28.3+/-4.1%), FGF-E(-) (29.7+/-3.3%), and LacZ-E(-) (28.1+/-2.5%) groups. Postmortem angiograms and histological analyses showed that vascularisation in the distal part of the skin flap was significantly increased in the FGF-E(+) group compared with the other groups. CONCLUSION: These findings suggested that gene delivery of bFGF to the recipient bed muscles enhanced vascularity and viability of an ischemic skin flap, and that plasmid-based gene delivery with electroporation was a suitable delivery method. PMID- 15897038 TI - Histological evaluation of Permacol as a subcutaneous implant over a 20-week period in the rat model. AB - This study assessed the suitability of Permacol (a porcine derived, isocyanate cross linked collagen based biomaterial) as an alternative to autologous tissue in soft tissue reconstruction. The Sprague-Dawley rat was used as a model for subcutaneous implantation over a 20 week period and comparison made with two other porcine biomaterials (small intestinal submucosa and glycerol treated ethylene oxide sterilised porcine dermis). Implants were scored histometrically on the degree of acute inflammation, chronic inflammation, fibrosis and stromal response. The vascularity and percentage composition of collagen within Permacol were assessed by stereology and seescan image analysis, respectively. In general terms, Permacol was well tolerated as a subcutaneous implant, with only a minor chronic inflammatory response remaining after a 20 week period of implantation. There was evidence of collagen degradation during this period and vascular ingrowth into Permacol was limited. Permacol has the potential for a broad range of applications in plastic surgery, but may benefit from modification to promote a more rapid degree of vascularisation. PMID- 15897039 TI - An experimental model of peripheral nerve adhesion in rabbits. AB - We studied the effects of peripheral nerve adhesion in a rabbit sciatic nerve model. After nerve exposure, its adventitial layer was sutured with 8-0 nylon to the nerve bed, which had been cauterised to promote adhesion. Nerve kinematics, electrophysiology, blood flow and histology were assessed. Rabbits in which Fontana's bands were visible as normal through the epineurium, classified as a nonadhesion group (group I), lacked intraneural fibrosis. In this group, nerve conduction and nerve blood flow were well maintained. Rabbits in which Fontana's bands could not be seen were classified as the adhesion group (group II). This group was classified into two levels pathologically; thickening of epineurium and perineurium was observed but no endoneurial fibrosis (group IIa), and endoneurial fibrosis (Wallerian degeneration, myelin sheath thinning and fibrosis between nerve fibers) was noted (group IIb). Compound muscle action potentials (CMAP) were reduced in amplitude and blood flow was significantly decreased at adhesion sites in group IIb. In conclusion, adhesion of peripheral nerve to surrounding tissues results in fibrosis in the nerve that contributes to peripheral nerve dysfunction. PMID- 15897040 TI - The use of thoracodorsal nerve transfer in restoration of irreparable C5 and C6 spinal nerve lesions. AB - There are only a few reports on the use of thoracodorsal nerve (TDN) transfer to the musculocutaneous or axillary nerves in cases of directly irreparable brachial plexus injuries. In this study, we analysed outcome and time-course of recovery in correlation with recipient nerves and type of nerve transfer (isolated or in combination with other collateral branches) for 27 patients with transfer to the musculocutaneous or axillary nerves. Using this nerve as donor, we obtained useful functional recovery in all 12 cases for the musculocutaneous nerve, and in 14 (93.3%) of 15 nerve transfers for the axillary nerve. Although, we found no significant statistical difference between analysed patients according to the percentage of recoveries and mean values, we established a better quality and shorter time of recovery for the musculocutaneous nerve. According to obtained results, we consider that transfer may be a valuable method in reconstruction after directly irreparable C5 and C6 spinal nerve lesions. PMID- 15897041 TI - Fish vaccine injection injuries of the hand. AB - High pressure injection of oil-based substances can cause devastating injuries of the hand and the necessity for urgent surgical debridement has been well established. We present three cases of injection injury caused by vaccines used in the fish farming industry. Patients presented with pain, swelling and lymphangitis. The marked vascular changes often associated with such injuries were absent. All patients were treated with intravenous antibiotics combined with early surgical debridement and irrigation. We report full recovery in all three patients. PMID- 15897042 TI - An anatomical study and clinical cases of 'super-thin flaps' with transverse cervical perforator. AB - We present a microangiogram study and clinical cases of super-thin flaps based on a transverse cervical perforator. This flap is the first to use the perforator of the superficial branch of the transverse cervical artery. This flap is more useful for providing colour and texture matches than a skin graft, and it is easier to harvest the flap than a free flap because it is a kind of skin flap. PMID- 15897043 TI - Aesthetic surgery techniques after excision of dermatofibrosarcoma protuberans: a case report. AB - Dermatofibrosarcomata protuberans are rare locally aggressive tumours that have a very high recurrence rate in the absence of wide resection. Normally, when the area to be covered is exceptionally large or when skin grafting is not possible because of the location, pedicle or free flaps are used. We report one case of wide excision where the opportunity arose to apply aesthetic surgery techniques combining bilateral superiorly based mammaplasty and reverse abdominoplasty. PMID- 15897044 TI - Superficial acral fibromyxoma. AB - Superficial acral fibromyxoma is a rare soft tissue tumour affecting the digits, particularly the nail bed region. The condition was first described in 2001. We report the case of a 71-year-old man who presented with a long standing history of a lump affecting the pulp of the his right ring finger. This was surgically excised and subsequent pathological analysis confirmed the lesion to have been a superficial acral fibromyxoma. We use this case to highlight the features of this rare clinical entity, which has never previously been described in the surgical literature. PMID- 15897045 TI - Sciatic nerve enlargement in the Klippel-Trenaunay-Weber syndrome. AB - The case of a 35-year-old woman with Klippel-Trenaunay-Weber syndrome (KTWS) showing clinical symptoms of a peroneal nerve lesion is presented. An immense nerve enlargement along most of the sciatic, peroneal and tibial nerve was found to be due to a lipoma arising from the epi- and perineurium. Treatment consisted of extensive microsurgical neurolysis and excision of the tumor resulting in decompression of the affected nerves. Although rare, a perineural lipoma should be kept in mind in patients with KTWS showing neurological abnormalities. PMID- 15897046 TI - Actinomycosis complicating fibula flap mandible reconstruction: a report of two cases. AB - Two patients undergoing fibula flap mandible reconstruction developed chronic intraoral wounds and salivary fistulae. After initial attempts at salvage, tissue biopsies demonstrated Actinomycosis infection. With antibiotic treatment and debridement, one reconstruction was salvaged while one was lost. Actinomycosis infection should be considered a possible agent in chronic wounds complicating mandible reconstructions with microsurgical flaps. PMID- 15897047 TI - Two flaps and Z-plasty technique for correction of longitudinal ear lobe cleft. AB - Various surgical techniques have been reported for the correction of congenital ear lobe deformities. Our method, the two-flaps-and-Z-plasty technique, for correcting the longitudinal ear lobe cleft is presented. This technique is simple and easy to perform. It enables us to keep the bulkiness of the ear lobe with minimal tissue sacrifice, and to make a shorter operation scar. The small Z plasty at the free ear lobe margin avoids notching deformity and makes the shape of the ear lobe smoother. The result is satisfactory in terms of matching the contralateral normal ear lobe in shape and symmetry. PMID- 15897048 TI - The intrinsic neuroadipofascial pedicled fasciocutaneous flap. PMID- 15897049 TI - An unusual complication of penile piercing. PMID- 15897050 TI - Eye protection for plastic surgeons. PMID- 15897051 TI - The story of 'the wiggly worm'. PMID- 15897052 TI - Wriggling to freedom (from pain). PMID- 15897053 TI - Characteristic adsorption functions and the surface structure of solid adsorbents. AB - A thermodynamic model of gas/solid adsorption has been constructed from two elements. The first is the original Gibbs equation. The second consists of functions psi(Theta) or psi(P) that are calculable from measured isotherms. Based on this model the characteristic adsorption functions (CAFs) were defined and calculated. The CAFs, which concentrate into one function all measured isotherms having the same change in relative free energy of the surface, are very sensitive to the structure of the adsorbents. This statement was tested with nitrogen isotherms measured at 77 K on well-characterized chemically/physically treated activated carbons prepared from poly(ethylene terephthalate). Changes in the surface structure were followed by small angle X-ray scattering (SAXS). This experimental approach made it possible to observe the correspondence between structural changes and the CAF. PMID- 15897054 TI - Mechanisms and kinetics of trisodium 2-hydroxy-1,1'-azonaphthalene-3,4',6 trisulfonate adsorption onto chitosan. AB - Chitosan, a naturally abundant biopolymer, has widely been studied for metal adsorption from various solutions, but the extension of chitosan as an adsorbent to remove organic substances from water and wastewater has seldom been explored. In this study, the adsorption of an azo dye, trisodium 2-hydroxy-1,1' azonaphthalene-3,4',6-trisulfonate (1), from aqueous solution onto the various degrees of deacetylated chitosan has been investigated. Equilibrium studies have been carried out to determine the capacity of chitosan for dye. The experimental data were analyzed using two isotherm correlations, namely, Langmuir and Freundlich equations. The linear correlation coefficients were determined for each isotherm and the Langmuir provided the best fit. The experimental adsorption isotherms were perfectly reproduced in the simulated data obtained from numerical analysis on the basis of the Langmuir model and the isotherm constants. Adsorption of (1) onto the chitosan flakes was found to be strongly depending on degrees of deacetylation in chitosan and temperatures. Significant amounts of (1) were adsorbed by chitosan 8B (higher degree of deacetylated chitosan), but the adsorption capacity was reduced remarkably with increasing solution temperatures. Thermodynamic parameters such as change in free energy (DeltaG), enthalpy (DeltaH), and entropy (DeltaS) were also determined. In addition, kinetic study indicated that the adsorption process mechanisms were both transport- and attachment-limited. PMID- 15897055 TI - Kinetic and thermodynamic studies of boron removal by Siral 5, Siral 40, and Siral 80. AB - In this study, adsorption of boron onto Siral 5, Siral 40, and Siral 80 samples was studied in a batch system with a function of temperature and contact time. The results of adsorption studies indicate that adsorption process was well described with the Freundlich equation and Dubinin-Radushkevich (DR) equation. The Langmuir model does not appear to fit the adsorption better than Freundlich or DR. It has also been found that the magnitudes of free energies are less than 8 kJ mol(-1) and within the energy range of physical adsorption. The intraparticle diffusion model and pseudo-first-order and pseudo-second-order kinetic model were tested to find out rate constants of adsorption. From kinetic experiments, it is realized that boron adsorption onto Siral samples obeyed the pseudo-second-order kinetic model. The thermodynamic studies showed that adsorption process was not spontaneous nature and endothermic. It could also be added that the randomness increased during the adsorption. PMID- 15897056 TI - Effect on uptake of heavy metal ions by phosphate grafting of allophane. AB - The effect of phosphate grafting on the uptake of various heavy metal ions was investigated using allophane, a clay mineral having a chemical composition of 1 2SiO2Al2O3.5-6H2O with hollow nanoparticles 3.5-5 nm in size. Phosphate grafting was performed on allophane with an atomic ratio Si/Al=0.85 using the orthophosphates (NH4)2HPO4, Na2HPO4 and CaHPO4 and the triphosphate Na5P3O10. The cations in the phosphates were partially retained in the samples, along with the grafted phosphate. Uptake experiments were performed with various heavy metal ions (Cu2+, Zn2+, Ni2+, Co2+, and Mn2+) and alkaline earth ions (Mg2+ and Ca2+) using a batch method. With increasing amounts of grafted phosphate, the Cu2+ uptake capacity increases to about double of that of ungrafted allophane. Although part of the grafted phosphate is released during the uptake experiments, this can be largely suppressed by heat-treating the samples at 200-500 degrees C. The Cu2+ uptake ability of the (NH4)2HPO4-grafted sample showed a steep decrease with higher heating temperature according to the thermal decomposition of NH4+. The ratios of cations released from the samples to cations removed from solution (2Na/Cu and Ca/Cu) are close to unity. From these results, the replacement reaction is thought to be the main uptake mechanism of heavy metal ions in the present samples. The uptake abilities of the present samples for various cations are explained with respect to the solubilities of the corresponding metal phosphates. PMID- 15897057 TI - Adsorption of basic dyes from aqueous solution onto pumice powder. AB - The adsorption of methylene blue and crystal violet on pumice powder samples of varying compositions was investigated using a batch adsorption technique. The effects of various experimental parameters, such as adsorbent dosage, initial dye concentration, and contact time, were also investigated. The extent of dye removal increased with decreased initial concentration of the dye and also increased with increased contact time and amount of adsorbent used. Adsorption data were modeled using the Freundlich adsorption isotherm. The adsorption kinetic of methylene blue and crystal violet could be described by the pseudo second-order reaction model. PMID- 15897058 TI - Analysis of application of Langmuir isotherm to heterogeneous systems: high pressure conditions. AB - In this paper, an adsorption model that is based on the statistical mechanics approach was applied to study the saturation phenomena in adsorption to calculate the minimum pressure needed to attain the complete surface coverage on a physical adsorption. The fundamental integral equation, Theta(T)(Q)=integral N(Q)Theta(Q)dQ, for the calculation of the coverage degree of the surface was developed for an exponential distribution function, N(Q)=(m/RT)exp(-mQ/RT), and the representation of local adsorption sites is given by the Langmuir expression, Theta(Q)=b(0)C exp(Q/RT)/(1+b(0)C exp(Q/RT)). At high values of the pressure C, a solution of the fundamental integral equation was obtained by imposing the condition b(0)C>1. The expression for the saturation condition, b(0)C>m/(m+1), was obtained; that is, the saturation phenomenon is dependent on two parameters, correlated with the energetic heterogeneity and adsorption energy of the system. The pressure in the analysis of the cited expression shows that, for low m values (more heterogeneous systems), saturation is attained for b(0)C>>m, while for m congruent with 1 (more homogeneous systems), the saturation is attained for b(0)C>>0.5. PMID- 15897059 TI - Adsorption characteristics of various organic substances on the surfaces of tantalum, titanium, and zirconium. AB - Adsorption characteristics of carboxylic acids, amines, an octapeptide composed of four L-alanine and four L-aspartic acid residues (Peptide-A4D4), and beta lactoglobulin (beta-Lg) on tantalum (Ta), titanium (Ti), and zirconium (Zr) particles were examined at 30 degrees C and in some case, were compared with their adsorption onto SUS316L stainless steel particles (S6L). The adsorption isotherms on the Ta, Ti, and Zr particles could usually be expressed either by a Langmuir-type equation for reversible adsorption or by a modified Langmuir-type adsorption equation including terms for both reversible and irreversible adsorption. The adsorption equilibrium of benzoic acid, benzylamine, and m xylylenediamine on all the metal surfaces followed a Langmuir-type equation, while those of phthalic acid, mellitic acid, and Peptide-A4D4 could be fitted to the modified Langmuir-type adsorption equation. The adsorption characteristics of different adsorbates on the different surfaces were discussed particularly with reference to the pH dependencies of the q(irrev), q(rev), and K values and the electrostatic properties of the oxidized surface of the metal particles. Fourier transform infrared spectroscopic analyses using a reflection/absorption technique (RA-IR) indicated that phthalic acid and mellitic acid are adsorbed in similar adsorption states irrespective of the type of metal. beta-Lg was adsorbed onto the surfaces principally in an irreversible manner. The desorption behavior of beta-Lg from Ta, Ti, and S6L surfaces was examined, in order to evaluate the extent of interaction between beta-Lg and the metals. PMID- 15897060 TI - AAS, XRPD, SEM/EDS, and FTIR characterization of Zn2+ retention by calcite, calcite-kaolinite, and calcite-clinoptilolite minerals. AB - In this study, the sorption behavior of Zn2+ on calcite, kaolinite, and clinoptilolite, in addition to mixtures of calcite with kaolinite and clinoptilolite, was investigated at various loadings and mixture compositions using atomic absorption spectroscopy, scanning electron microscopy/energy dispersive X-ray spectroscopy, X-ray powder diffraction, and Fourier transform infrared techniques. According to the obtained results, within the experimental operating conditions, the sorption capacity was enhanced with increasing amount of calcite in both types of mixtures. Under neutral-alkaline pH conditions and high loadings, the order of Zn2+ retention was observed as calcite>clinoptilolite>kaolinite. The experiments on the retention of Zn2+ by pure calcite under conditions of oversaturation showed that the uptake process proceeds via an initial adsorption mechanism (possibly ion-exchange type) followed by a slower mechanism that leads to the overgrowth of the hydrozincite phase, Zn5(OH)6(CO3)2. PMID- 15897061 TI - Temperature-dependent intermolecular force measurement of poly(N isopropylacrylamide) grafted surface with protein. AB - We have investigated the temperature dependence of the intermolecular force between poly(N-isopropylacrylamide) (PNiPAM) grafted surface and bovine serum albumin (BSA) in phosphate buffer (pH 7.4) using atomic force microscopy at the nanonewton scale. These observations show that the interaction force is nearly zero below the phase transition temperature of PNiPAM and that it increases steeply during the phase transition. Since the PNiPAM chains are grafted onto the aminosilane (gamma-aminopropyltriethoxysilane)-treated silicon wafer, we measured the force-distance curve of BSA-immobilized tips for the bare and the aminosilane treated silicon wafer. These surfaces show no temperature dependence and their values are different from those of the PNiPAM-grafted surfaces at 30 degrees C. The results indicate that the measured adhesion force is between the PNiPAM grafted surface and the BSA-immobilized tip. Our studies on the intermolecular force between other surfaces (CH(3)- and COOH-terminated self-assembled monolayers) and the BSA-immobilized tip indicate that the variation in the intermolecular force between the PNiPAM surface and BSA with temperature can be attributed to the changes in the properties of the PNiPAM chains. From consideration of the PNiPAM phase transition mechanism, it is speculated that the intermolecular force between the PNiPAM-grafted surface and BSA would be affected by changes in the arrangement of the bound water molecules around the PNiPAM chain and by changes in the conformation (i.e., in the chain mobility) of the PNiPAM chain during the phase transition. PMID- 15897062 TI - Cell surface groups of two picocyanobacteria strains studied by zeta potential investigations, potentiometric titration, and infrared spectroscopy. AB - In order to clarify the role of picocyanobacteria in aquatic biogeochemical processes (e.g., calcite precipitation), cell surface properties need to be investigated. An experimental study of the cell surface characteristics of two Synechococcus-type unicellular autotrophic picocyanobacterial strains was carried out. One strain was isolated from Lake Plon and contained phycocyanin, the other strain came from Lago Maggiore and was rich in phycoerythrin. Potentiometric titrations were conducted to determine the different types of sites present on the bacteria cell walls. Infrared spectroscopy allowed characterization of the various functional groups (RNH(2), RCOOH, ROH, RPO(2)) and investigations of zeta potential provided insight into the isoelectrical points of the strains. Titrations reveal three distinct sites on the bacterial surfaces of phycocyanin- and phycoerythrin-rich strains with pK values of 4.8+/-0.3/5.0+/-0.2, 6.6+/ 0.2/6.7+/-0.4, and 8.8+/-0.1/8.7+/-0.2, corresponding to carboxyl, phosphate, and amine groups with surface densities of 2.6+/-0.4/7.4+/-1.6 x 10(-4), 1.9+/ 0.5/4.4+/-0.8 x 10(-4), and 2.5+/-0.4/4.8+/-0.7 x 10(-4) mol/g of dry bacteria. The deprotonation constants are similar to those of bacterial strains and site densities are also within an order of magnitude of other strains. The phycoerythrin-rich strain had a higher number of binding sites than the phycocyanin-rich strain. The results showed that picocyanobacteria may adsorb either calcium cations or carbonate anions and therefore strongly influence the biogeochemical cycling of calcite in pelagic systems. PMID- 15897063 TI - BSA adsorption on bimodal PEO brushes. AB - BSA adsorption onto bimodal PEO brushes at a solid surface was measured using optical reflectometry. Bimodal brushes consist of long (N=770) and short (N=48) PEO chains and were prepared on PS surfaces, applying mixtures of PS(29)-PEO(48) and PS(37)-PEO(770) block copolymers and using the Langmuir-Blodgett technique. Pi-A isotherms of (mixtures of) the block copolymers were measured to establish the brush regime. The isotherms of PS(29)-PEO(48) show hysteresis between compression and expansion cycles, indicating aggregation of the PS(29)-PEO(48) upon compression. Mixtures of PS(29)-PEO(48) and PS(37)-PEO(770) demonstrate a similar hysteresis effect, which eventually vanishes when the ratio of PS(37) PEO(770) to PS(29)-PEO(48) is increased. The adsorption of BSA was determined at brushes for which the grafting density of the long PEO chains was varied, while the total grafting density was kept constant. BSA adsorption onto monomodal PEO(48) and PEO(770) brushes was determined for comparison. The BSA adsorption behavior of the bimodal brushes is similar to the adsorption of BSA at PEO(770) monomodal brushes. The maximum of BSA adsorption at low grafting density of PEO(770) can be explained by ternary adsorption, implying an attraction between BSA and PEO. The contribution of primary adsorption to the total adsorbed amount is negligible. PMID- 15897064 TI - Interactions between dialkyldimethylammonium bromides (DXDAB) and sterols--a monolayer study. AB - Langmuir monolayers of cholesterol/ergosterol and dialkyldimethylammonium bromides (DXDABs) differing in alkyl chain length-14 (DTDAB), 16 (DHDAB), and 18 (DODAB)-spread at the air/water interface are examined. All the systems investigated are found to be nonideal and miscible. Negative values of the total free energy of mixing, proving film stability in the whole range of compositions and surface pressures, are observed for all the studied mixtures except for DTDAB/cholesterol. The strength of interactions, quantified with DeltaG(Exc) values, was found to be of the same order for mixtures of cholesterol/ergosterol and DHDAB or DODAB. Differences occurring for the mixtures of DTDAB with sterols indicate the affinity of DTDAB to ergosterol in contrast to cholesterol. PMID- 15897065 TI - The influence of Al(III) supersaturation and NaOH concentration on the rate of crystallization of Al(OH)3 precursor particles from sodium aluminate solutions. AB - The growth kinetics of colloidal Al(III)-containing particles (diameter<1000 nm), nucleated in optically clear, supersaturated sodium aluminate solutions as a precursor to Al(OH)(3) crystals, has been studied using dynamic light scattering. Two series of solutions were examined at 22 degrees C to determine the influence of Al(III) supersaturation and NaOH concentration on the initial particle growth behavior. One solution series consisted of solutions with constant Al(III) absolute supersaturation (DeltaC) of 1.48 M and [NaOH] range 1.83-4.00 M ([NaOH]/[Al(III)]=1.13-2.15) and Al(III) relative supersaturation (sigma)=3.86 10.36. The other solution series had a constant sigma of 7.55 and [NaOH] range of 1.50-4.27 M ([NaOH]/[Al(III)]=1.18-1.54) and DeltaC=0.86-3.19. The correlation between the initial particle growth rates and supersaturation (DeltaC or sigma) revealed marked anomalies over the entire supersaturation range studied. The growth rate remained substantially constant in the DeltaC range 0.86-2.55 M (for the constant sigma solution series), before increasing sharply upon a further increase of DeltaC beyond 2.55 M. The variation of the growth rate with sigma in the range 3.86-9.00 (for the constant DeltaC solution series) was remarkably weak, contrary to expectation. At higher sigma (>9.00), however, a marked increase in growth rate with increasing sigma was displayed. At constant DeltaC or sigma, the growth rate showed a strong variation with NaOH concentration, indicating that Na(+) and OH(-) species play a pivotal role in the Al(OH) precursor particles (nuclei) growth process. Furthermore, the kinetics of growth displayed by these nanosized particles are an order of magnitude slower than those observed for macroscopic gibbsite (gamma-Al(OH)(3)) crystals at similar supersaturations and temperature. The difference may be rationalized in terms of particle size and Al(OH)(3) dimorphic phase dependent solubility effects. An empirically adequate growth kinetics modeling was achieved when the growth rates were correlated with the Al(III) supersaturation (DeltaC or sigma) and the excess (free) NaOH concentration, rather than the former alone, as is commonly the case. A critical [NaOH]/[Al(III)] molar ratio of 1.27-1.35, below which the particle growth rate increased markedly and above which the rate was significantly reduced, was observed. This behavior is believed to be linked to solution speciation change that occurs at certain Al(III) and NaOH compositions. PMID- 15897066 TI - Effects of the water content on the growth rate of AgCl nanoparticles in a reversed micelle system. AB - The effects of water content on the growth rate and the final particle size of AgCl nanoparticles in a reversed micelle (RM) system of polyoxyethylene (6) nonylphenyl ether (NP-6)/water/cyclohexane were investigated using a double-jet technique, in which RM solutions of AgNO(3) and KCl were added concurrently to a RM solution containing the excess concentration of chloride ion. As a result, the particle growth rate and the final particle size at a constant Rw ( identical with[water]/[surfactant]) below 5 were found to be in excellent agreement with our theoretical prediction based on a dynamic Ostwald ripening mechanism governed by the overall solubility of the solid and the diffusivity of the reversed micelles, whereas the final particle size was far beyond the size of the water pool of a reversed micelle. Thus, the dramatic reduction of the particle size in the RM system can be explained by the drastic reduction of the overall solubility of the solid and the small diffusivity of the bulky reversed micelles as a carrier of silver ion, and not by the size of the water pool of a reversed micelle as conventionally explained. Some additional contribution of a coagulation process was also suggested in a high Rw range above 5. Significant coagulation of AgCl particles was observed in a RM system with AOT in place of NP 6 even under the standard conditions for the NP-6 system. PMID- 15897067 TI - Colloid stability of synthetic titania and the influence of surface roughness. AB - The colloid stability of synthetic titania particles was studied as a function of KCl concentration at pH values of 6.3, 6.7, and 8.4, using static light scattering to obtain stability ratios. Standard DLVO theory was then used to calculate the stability ratios as a function of salt concentration. Reasonable agreement between theory and experiment could only be obtained if an effective interaction radius, corresponding to surface asperities on the titania particles, was used in the calculation. High-resolution TEM images suggest that the effective interaction radius corresponds to the size of surface crystallites formed during synthesis. PMID- 15897068 TI - Preparation of large monodispersed spherical silica particles using seed particle growth. AB - To obtain large-sized, monodispersed spherical particles of silica by sol precipitation, a seed particle growth method was attempted. The formation of secondary particles during seed particle growth causing a multimodal distribution of particle size was suppressed via fine adjustment of the reaction conditions, such as TEOS, ammonia, and water concentrations, as well as operational conditions such as feeding time and agitation speed. Among the reaction conditions, an increase of TEOS concentration promoted secondary particle formation, resulting in bimodal particle distribution. However, secondary particle formation was depressed with increasing ammonia and water concentrations. In addition, long feeding time (low feed flow rate) and rigorous agitation significantly reduced secondary particle formation because they contributed to the slow generation of supersaturation and rapid seed particle growth, respectively. PMID- 15897069 TI - Ceramic nanomaterials from aqueous and 1,2-ethanediol supersaturated solutions at high temperature. AB - The aim of this study was the synthesis and physicochemical characterization of some nanosized hydroxides/oxides interesting in the field of ceramic materials with the aim of developing new materials for technological applications in the field of high-performance coatings. In particular, attention was focused on ZrO2nH2O, ZrO2, Zn(OH)2, ZnO, Al2O3nH2O, and Al2O3. The synthesis was carried out in 1,2-ethanediol at 150-160 degrees C or in water at 90 degrees C at a high degree of supersaturation in order to achieved nucleation rate much greater than the growth rate. The obtained ZrO2nH2O, Zn(OH)2, and Al2O3nH2O particles were peptized to eliminate possible agglomeration and then calcinated at a suitable temperature to allow the formation of the corresponding anhydrous oxides. The synthesized particles were characterized by scanning and transmission electron microscopy, X-ray diffractometry, and differential thermal analysis coupled with thermogravimetry and Fourier transform infrared spectroscopy. The study showed that all the products can be obtained in the nanostructured crystalline form; ZrO2 presented the interesting feature of different tetragonal/monoclinic ratios depending on the solvent used for the synthesis. Preliminary results on the potentialities of these ZrO2 particles as agents for ultrahard coatings on ceramic surfaces were encouraging and very promising. PMID- 15897070 TI - Influence of environmental stresses on stability of oil-in-water emulsions containing droplets stabilized by beta-lactoglobulin-iota-carrageenan membranes. AB - An oil-in-water emulsion (5 wt% corn oil, 0.5 wt% beta-lactoglobulin (beta-Lg), 0.1 wt% iota-carrageenan, 5 mM phosphate buffer, pH 6.0) containing anionic droplets stabilized by interfacial membranes comprising of beta-lactoglobulin and iota-carrageenan was produced using a two-stage process. A primary emulsion containing anionic beta-Lg coated droplets was prepared by homogenizing oil and emulsifier solution together using a high-pressure valve homogenizer. A secondary emulsion containing beta-Lg-iota-carrageenan coated droplets was formed by mixing the primary emulsion with an aqueous iota-carrageenan solution. The stability of primary and secondary emulsions to sodium chloride (0-500 mM), calcium chloride (0-12 mM), and thermal processing (30-90 degrees C) were analyzed using zeta potential, particle size and creaming stability measurements. The secondary emulsion had better stability to droplet aggregation than the primary emulsion at NaCl Cr(3+)>Mn(2+). The equilibrium data fit well with the Langmuir and Freundlich models and confirm the affinity order of the soil sample for these metals. These adsorption data are combined with EPR spectroscopy to obtain structural information about the surface complexes formed. Iron is held in inner sphere complexes. Manganese is simultaneously held in outer- and inner-sphere complexes. Due to poor resolution, chromium was not detected by EPR and thus it is impossible to infer coordination sphere and coordination number. Iron and manganese are in an octahedral environment. PMID- 15897077 TI - Synthesis and catalytic activity of gold-silver binary nanoparticles stabilized by PAMAM dendrimer. AB - Gold-silver binary nanoparticles, which feed atomic ratios of gold to silver were 3:1, 1:1, and 1:3, were prepared. These particles were stabilized by amine terminated (generation (G) 3.0 and 5.0) and carboxyl-terminated (G 3.5 and 5.5) poly(amidoamine) (PAMAM) dendrimers in water. UV-vis spectra indicate that the particles are not mere physical mixtures of monometallic particles or core/shell type but alloy. According to transmission electron microscope (TEM) observation, the mean diameters of the particles were 7-10 nm for silver particles and 3-4 nm for both gold and alloy particles, respectively. Catalytic activities for reduction of p-nitrophenol were investigated by monitoring the absorbance at 400 nm during the reaction. They were proportional to the feed ratio of gold in the particles and showed a maximum at the ratio of Au:Ag=3:1. PMID- 15897078 TI - Surface potential at the hematite-water interface. AB - Construction of a metal oxide electrode enabling measurement of surface potential is described. The electrode was made using a hematite monocrystal, which avoids the problems arising from the possible porosity of the oxide layer. The potential of this electrode was measured as a function of pH. The hematite electrode provides reproducible results, especially in the acidic region. Surface potentials were calculated from electrode potentials using the electrokinetic isoelectric point. The slope of surface potential with respect to pH was found to be lower than the Nernstian, especially in the basic region. The effect was more pronounced at higher ionic strengths. It was shown how the measurement of surface potential can help to interpret the equilibrium data and evaluate the choice of a theoretical model describing the interfacial equilibrium at the metal oxide-water interface. PMID- 15897079 TI - Influence of relative humidity on electrical properties of alpha-Al2O3 powders: resistivity and electrochemical impedance spectroscopy. AB - The influence of humidity on the electrical properties of alpha-Al2O3 powders has been investigated using adsorption isotherms, DC resistivity, and electrochemical impedance spectroscopy. Samples of two alpha-Al2O3 particle sizes were examined, both individually and mixed together. The results show that the grain-bed resistivity decreases with humidity, whereas the grain capacitance is almost constant. The resistivity difference between the two particle sizes is of several orders of magnitude, while the capacitance values are not very different. These results are interpreted in terms of the layer-by-layer growth of water adsorbed on the grain surfaces. The first, more tightly bound adsorbed layer does not provoke the same effects as those layers adsorbed at higher relative humidity. PMID- 15897080 TI - Photocatalytic degradation and adsorption of 2-naphthol on suspended TiO2 surface in a dynamic reactor. AB - The photocatalytic oxidation of 2-naphthol has been investigated at room temperature in a dynamic photoreactor with system UV/O2 (air) and aqueous suspension of titanium dioxide TiO2 irradiated under a variety of conditions. The kinetics of disappearance of pollutant were affected by several operating parameters such as TiO2 mass, concentration of the substrate and reaction pH. The experiments were measured by high performance liquid chromatography. A Langmuir Hinshelwood model was found to be accurate for photocatalytic degradation and indicates that adsorption of the solute on the surface of semiconductor particles plays a role in photocatalytic reaction. PMID- 15897081 TI - Synthesis and characterization of ordered hexagonal and cubic mesoporous tin oxides via mixed-surfactant templates route. AB - Ordered hexagonal and cubic mesoporous tin oxides were synthesized for the first time in the presence of mixed cationic and neutral surfactants (a mixture of cetyltrimethylammonium bromide cationic surfactant and dodecylamine neutral surfactant) with different alkali and simple inorganic precursors at room temperature. In the synthesis systems, the dodecylamine neutral surfactant may function as a polar organic cosolvent and cosurfactant. The formation of the tin oxide mesostructured material was proposed to be due to the presence of hydrogen bonding interactions between the supramolecular template and inorganic precursors Sn4+ and OH-, which were assumed to self-assemble around the cationic surfactant molecules. The materials are characterized by X-ray powder diffraction, transmission electron microscopy, thermogravimetric analysis, and N2 adsorption/desorption isotherm. The surface areas of materials evaluated from the N2 sorption isotherms are about 248 m(2)/g for hexagonal mesoporous tin oxide (SnH) and 281 m(2)/g for cubic mesoporous tin oxide (Sn-C) for calcination at 350 degrees C. PMID- 15897082 TI - Roughness and hydrophobicity studies of nanofiltration membranes using different modes of AFM. AB - Determination of the surface roughness by AFM is crucial to the study of particle fouling in nanofiltration. It is, however, very difficult to compare the different roughness values reported in the literature because of a lack in uniformity in the methods applied to determine surface roughness. AFM is used in both noncontact mode and tapping mode; moreover, the size of the scan area is highly variable. This study compares, for six different nanofiltration membranes (UTC-20, N30F, Desal 51HL, Desal 5DL, NTR7450, NF-PES-10), noncontact mode AFM with tapping mode AFM for several sizes of the scan area. Although the absolute roughness values are different for noncontact AFM and tapping mode AFM, no difference is found between the two modes of AFM in ranking the nanofiltration membranes with respect to their surface roughness. NTR 7450 and NF-PES-10 are the smoothest membranes, while the roughest surface can be found with Desal 51HL and Desal 5DL. UTC-20 and N30F are characterized by an intermediate roughness value. An increase in roughness with increasing scan area is observed for both AFM modes. Larger differences between the roughnesses of the membranes are obtained with tapping mode AFM because of the tapping of the tip on the surface. Phase imaging is an extension of tapping mode AFM, measuring the phase shift between the cantilever oscillation and the oscillation of the piezo driver. This phase shift reflects the interaction between the cantilever and the membrane surface. A comparison with contact angle measurements proves that a small phase shift corresponds to a large contact angle, representing a hydrophobic membrane surface. PMID- 15897083 TI - Electrodialysis of calcium and carbonate high concentration solutions and impact on composition in cations of membrane fouling. AB - Fouling, which is the accumulation of undesired solid materials at the phase interfaces of permselective membranes, is one of the major problems in electrodialysis. The objectives of the present work were to investigate the effect of the composition in calcium and carbonate of a model solution to be treated by conventional electrodialysis on their migration kinetics and the composition in cations of the membrane fouling. In the absence of sodium carbonate in the solution, no fouling was visually observed on anion-exchange membranes (AEM) and fouling was observed only at 1600 mg/L CaCl2 on cation exchange membrane (CEM), while at only 800 mg/L CaCl2 with sodium carbonate, a deposit was observed on both membranes. This difference could be explained by the fact that carbonate has a high buffer capacity, and the time to reach pH 4.0 was then longer than the one without carbonate. Consequently, the migration of the ionic species was carried out over a longer period of time during ED treatment with sodium carbonate addition and in extent the demineralization rates were higher: 43 vs 86%. For treatment with sodium carbonate and 1600 mg/L CaCl2, the higher migration during ED treatment, increased the concentration of calcium, from 14.24 to 93.38 mg/g dry membrane and from 0.74 to 10.27 mg/g dry membrane for CEM and AEM, respectively. Due to the basic pH on the side of the membrane in contact with the NaCl solution, the calcium would precipitate to form calcium hydroxide on CEM while the calcium migrated through the CEM was blocked by the AEM where it formed another fouling. PMID- 15897084 TI - A finite element based algorithm for determining interfacial tension (gamma) from pendant drop profiles. AB - This paper introduces a robust algorithm to determine the interfacial tension (gamma) from pendant drop profiles using the Galerkin finite element method (gamma-PD-FEM) to solve the axisymmetric form of the Young-Laplace (YL) equation. In this algorithm, the theoretical profiles are generated by solving the spherical coordinate form of the YL equation. gamma-PD-FEM also solves for the parameter estimates by minimizing the difference between the theoretical and experimental surface functions, f(theta). This technique is compared to the widely used method of converting the YL equation to the three arc length-based (ALB) first-order ODEs developed by Bashforth and Adams (BA) in 1883, or as denoted in this paper, the gamma-PD-BA method. The drop apex is the initial condition for the gamma-PD-BA algorithm and the integration is terminated at a specified location along the drop profile. In contrast to techniques based on the BA approach, computation of the theoretical drop profile in gamma-PD-FEM is obtained from a second-order ordinary differential equation and requires boundary conditions at the drop apex and at the contact line of the drop to the nozzle. By incorporating both boundary conditions into the problem formulation, the algorithm can also determine if the drop shape is at static equilibrium. Results to be presented include an outline of the computer algorithm, and comparison of gamma values obtained from the gamma-PD-FEM and the traditional gamma-PD-BA method using simulated and experimental drop profile data sets. PMID- 15897085 TI - Impact of droplets onto inclined surfaces. AB - Drop impacts onto dry walls and liquid films at low impact angles and low normal Weber numbers are experimentally investigated. Measurements were performed using a high spatial resolution CCD camera and short exposure times, yielding both qualitative and quantitative information about the impact. Whereas a droplet generally deposits on the surface for high impact angles, a rebound can occur at lower angles and for smooth or wetted surfaces. No rebound is observed for rough surfaces. A low viscous liquid (water) will either rebound or deposit on smooth or wetted surfaces. A high viscous liquid (glycerin) may also disjoin into two droplets, depending on the impact angle. A correlation is presented for the size of the secondary droplet. A further correlation quantifies the critical impact angle at which rebounding first occurs in terms of the normal Weber number. PMID- 15897086 TI - A robust algorithm for the simultaneous parameter estimation of interfacial tension and contact angle from sessile drop profiles. AB - The pendant and sessile drop profile analysis using the finite element method (PSDA-FEM) is an algorithm which allows simultaneous determination of the interfacial tension (gamma) and contact angle (theta(c)) from sessile drop profiles. The PSDA-FEM algorithm solves the nonlinear second-order spherical coordinate form of the Young-Laplace equation. Thus, the boundary conditions at the drop apex and contact position of the drop with the substrate are required to solve for the drop profile coordinates. The boundary condition at the position where the drop contacts the substrate may be specified as a fixed contact line or fixed contact angle. This paper will focus on the fixed contact angle boundary condition for sessile drops on a substrate and how this boundary condition is used in the PSDA-FEM curve-fitting algorithm. The PSDA-FEM algorithm has been tested using simulated drop shapes with and without the addition of random error to the drop profile coordinates. The random error is varied to simulate the effect of camera resolution on the estimates of gamma and theta(c) values obtained from the curve-fitting algorithm. The error in the experimental values for gamma from sessile drops of water on acrylic and Mazola corn oil on acrylic falls within the predicted range of errors obtained for gamma values from simulated sessile drop profiles with randomized errors that are comparable in magnitude to the resolution of the experimental setup. PMID- 15897087 TI - Microscopic treatment of a barrel drop on fibers and nanofibers. AB - The microscopic approach of Berim and Ruckenstein (J. Phys. Chem. B 108 (2004) 19330, 19339) regarding the shape and stability of a liquid drop on a planar bare solid surface is extended to a liquid barrel drop on the bare surface of a solid cylinder (fiber) of arbitrary radius. Assuming the interaction potentials of the liquid molecules between themselves and with the molecules of the solid of the London-van der Waals form, the potential energy of a liquid molecule with an infinitely long fiber was calculated analytically. A differential equation for the drop profile was derived by the variational minimization of the total potential energy of the drop by taking into account the structuring of the liquid near the fiber. This equation was solved in quadrature and the shape and stability of the barrel drop were analyzed as functions of the radius of the fiber and the microscopic contact angle theta(0) which the drop profile makes with the surface of the fiber. The latter angle is dependent on the fiber radius and on the microscopic parameters of the model (strength of the intermolecular interactions, densities of the liquid and solid phases, hard core radii, etc.). Expressions for the evaluation of the microcontact angle from experimentally measurable characteristics of the drop profile (height, length, volume, location of inflection point) are obtained. All drop characteristics, such as stability, shape, are functions of theta(0) and a certain parameter a which depends on the model parameters. In particular, the range of drop stability consists of three domains in the plane theta(0)-a, separated by two critical curves a=a(c)(theta(0)) and a=a(c1)(theta(0)) [a(c)(theta(0))h(m1) cannot exist, whereas in the third domain (between those curves) the drop can have values of h(m) either smaller than h(m1) or larger than h(m2), where h(m2)>h(m1) is a second critical height. For sufficiently large fiber radii, R(f)1 >/= microm, the critical curves almost coincide and only two domains, the first and the second, remain. The smaller the radius, the larger is the difference between the critical curves and the larger is the second domain of drop stability. The shape of the drop depends on whether the point (theta(0),a) on the theta(0)-a plane is far from the critical curve or near it. In the first case the drop profile has generally a large circular part, while in the second case the shape is either almost planar or contains a long manchon that is similar to a film on the fiber. PMID- 15897088 TI - Co-solvent effects on drag reduction, rheological properties and micelle microstructures of cationic surfactants. AB - Some quaternary cationic surfactants, when mixed with a counterion, are known to self-assemble into threadlike micelles in water. Such behavior causes drastic changes in rheological properties of even very dilute solutions, allowing them to be used as drag reducing agents (DRA) in turbulent pipe flow circulating systems, such as district cooling/heating systems. Surfactant self-assembly is a physicochemical phenomenon whose character depends on surfactant nature and concentration, nature of the solvent, temperature and type and concentration of counterions. This study investigates drag reduction (DR) and rheological properties of two cationic surfactants, Ethoquad O/12 (oleyl bis(hydroxyethyl)methylammonium chloride) and Ethoquad O/13 (oleyl tris(hydroxyethyl) ammonium acetate), with excess salicylate counterion (NaSal), in mixed solvents containing 0 to 28 wt% ethylene glycol (EG) and water. The addition of EG to the solvent had greater effects on solutions' DR ability, shear viscosity, apparent extensional viscosity and viscoelasticity at 25 degrees C than at approximately 0 degrees C. Cryo-TEM images show threadlike micelle in these systems. DR at low temperatures in solutions containing moderate amount of EG can be utilized in a new approach to energy saving in district cooling systems using EG-water based mixtures as the cooling fluids. PMID- 15897089 TI - Foams and foam films stabilized by CnTAB: influence of the chain length and of impurities. AB - Quantitative comparison of foam films and the corresponding foams is very demanding. One problem is the fact that investigations of foam films are usually performed at constant capillary pressures P, whereas in foams P is a function of the height of the foam column. A way out of this dilemma is to examine films and foams at the same P. The method of choice for the foam films is the thin film pressure balance (TFPB), whereas the corresponding investigation of foams is based on the foam pressure drop technique (FPDT). An extensive TFPB study on foam films stabilized by the cationic alkyltrimethylammonium bromides C(n)TAB with n=10, 12, 14, and 16 was performed by Bergeron. For this series a steep increase of the foam film stability was observed when the chain length was increased from n=12 to n=14. Moreover, the influence of impurities was found to be limited to the films stabilized by C(12)TAB. In order to study the correlation between the properties of films and foams, the present study deals with the respective foam properties investigated with the FPDT. It was found that both the steep increase in the film stability and the influence of impurities are also reflected in the properties of the foam. PMID- 15897090 TI - Marangoni effects in aqueous polypropylene glycol foams. AB - The foam behavior of three polypropylene glycols covering the molecular weight range between 192 and 725 g/mol has been examined. Static and dynamic surface tension data, as well as bubble size distribution and retention time in the foam, were incorporated into a simple model of foam stability. The latter clearly indicates that surface tension differences between the plateau border and lamellar region adjacent to the bubble surface are the dominant factor in controlling foamability, causing liquid flow in the direction opposite to liquid drainage, a process termed the Marangoni effect. PMID- 15897091 TI - Oil coating of hydrophobic surfaces from aqueous media: formation and kinetic study. AB - We perform oil coating of hydrophobic solid surfaces via aqueous media, from emulsions, and under the presence of a shear flow. The principle of such coating is based on the use of a system at the limit of aggregation to give rise to adhesion, with asymmetrical interfaces (oil droplet/water and solid surface/water) in order to favor the oil/surface adhesion in comparison to the oil/oil adhesion. This way, droplets stick to the solid substrate, whereas they are stable and homogeneously dispersed in the bulk. We have realized coatings from two systems of emulsions made of a mixture of hydroxy-terminated silicone oil and classical silicone oil and a mixture of sunflower oil and mineral oil. The kinetics of the coating is described by a Langmuir model where the adhesion between the oil particle and the surface is modeled as a first-order reaction. The resulting coatings are formed of oil droplets uniformly covering the solid surface. The coating density can vary with the nature of the experimental systems. PMID- 15897092 TI - Properties of mixed micelles of cationic gemini surfactants and nonionic surfactant triton X-100: effects of the surfactant composition and the spacer length. AB - The mixed micelles of cationic gemini surfactants C12C(S)C12Br2 (S=3, 6, and 12) with the nonionic surfactant Triton X-100 (TX100) have been studied by steady state fluorescence, time-resolved fluorescence quenching, electrophoretic light scattering, and electron spin resonance. Both the surfactant composition and the spacer length are found to influence the properties of mixed micelles markedly. The total aggregation number of alkyl chains per micelle (N(T)) goes through a minimum at X(TX100)=0.8. Meanwhile, the micropolarity of the mixed micelles decreases with increasing X(TX100), while the microviscosity increases. The presence of minimum in N(T) is explained in terms of the competition of the reduction of electrostatic repulsion between headgroups of cationic gemini surfactant with the enhancement of steric repulsion between hydrophilic headgroups of TX100 caused by the addition of TX100. The variations of micropolarity and microviscosity indicate that the incorporation of TX100 to the gemini surfactants leads to a more compact and hydrophobic micellar structure. Moreover, for the C12C3C12Br2/TX100 mixed micelle containing C12C3C12Br2 with a shorter spacer, the more pronounced decrease of N(T) at X(TX100) lower than 0.8 may be attributed to the larger steric repulsion between headgroups of TX100. Meanwhile, the increase of microviscosity and the decrease of micropolarity are more marked for the C12C12C12Br2/TX100 mixed micelle, owing to the looped conformation of the longer spacer of C12C12C12Br2. PMID- 15897093 TI - Effect of glycerol on micelle formation by ionic and nonionic surfactants at 25 degrees C. AB - The effect of glycerol on the micellization of the cationic surfactant cetyltrimethylammonium bromide (CTAB) and of the ethoxylated nonionic surfactant Brij 58 has been investigated by various experimental techniques. For both surfactants the critical micellar concentration (cmc), determined by surface tension measurements, is almost unaffected by the presence of glycerol in the mixture; only at high glycerol concentrations (>/=20% w/w) does the cmc significantly increase. The area per surfactant molecule at the air-solution interface, A, increases with increasing glycerol weight percentage, w(g). Fluorescence quenching measurements indicate that the presence of glycerol induces a lowering of the aggregation number of both surfactants. The glycerol intradiffusion coefficient has been measured by the pulsed-gradient spin-echo NMR technique as a function of glycerol content at constant surfactant concentration. It is almost unaffected by the presence of the surfactants, indicating that no direct glycerol-surfactant interaction occurs in the mixture. The surfactant intradiffusion coefficient has been also measured. In the case of CTAB, it increases with increasing glycerol concentration, a reflection of the decreased aggregation number. For Brij 58, in spite of the lowering of the aggregation number, the surfactant intradiffusion coefficient decreases with increasing glycerol concentration, suggesting an increase of the intermicellar interaction. The experimental evidence shows that for both surfactants the micellization is affected by the presence of glycerol through an indirect, solvent-mediated mechanism. In the case of CTAB, the main effect of glycerol is a lowering of the medium dielectric constant, which enhances the electrostatic interactions in solution. In the case of Brij 58, the results can be interpreted in terms of a salting-out effect according to which glycerol competes with the surfactant for water molecules, causing a dehydration of the surfactant ethoxylic headgroup. PMID- 15897094 TI - Aggregation behavior of hexadecyltrimethylammonium surfactants with various counterions in aqueous solution. AB - Both thermodynamic and microenvironmental properties of the micelles for a series of cationic surfactants hexadecyltrimethylammonium (C16TAX) with different counterions, F-, Cl-, Br-, NO3-, and (1/2)SO4(2-), have been studied. Critical micelle concentration (CMC), degree of micelle ionization (alpha), and enthalpy of micellization (DeltaH(mic)) have been obtained by conductivity measurements and isothermal titration microcalorimetry. Both the CMC and the alpha increase in the order SO4(2-) < NO3- < Br- < Cl- < F-, consistent with a decrease in binding of counterion, except for the divalent anion sulfate. DeltaH(mic) becomes less negative through the sequence NO3- < Br- < Cl- < F- < SO4(2-), and even becomes positive for the divalent sulfate. The special behavior of sulfate is associated with both its divalency and its degree of dehydration. Gibbs free energies of micellization (DeltaG(mic)) and entropies of micellization (DeltaS(mic)) have been calculated from the values of DeltaH(mic), CMC, and alpha and can be rationalized in terms of the Hofmeister series. The variations in DeltaH(mic) and DeltaS(mic) have been compared with those for the corresponding series of gemini surfactants. Electron spin resonance has been used to assess the micropolarity and the microviscosity of the micelles. The results show that the microenvironment of the spin probe in the C16TAX surfactant micelles depends strongly on the binding of the counterion. PMID- 15897095 TI - Electrokinetic phenomena at grafted polyelectrolyte layers. AB - During the last decades the electrokinetic theory of Smoluchowski (Z. Phys. Chem. 92 (1918) 129) was extended to be applicable for soft surfaces (grafted polyelectrolyte layers (PL), biological and artificial membranes, etc.) by either using the Debye approximation or numerical solutions. In the theory of Ohshima (Colloids Surf. A 103 (1995) 249) the nonlinearized Poisson-Boltzmann (PB) equation for thick and uniform PL is solved analytically and a general hydrodynamic equation is derived in an integral form. These advantages in the theory of Ohshima provided a base for the further development of a generalized electrokinetic theory for soft surfaces. In his theory the final equation for the electroosmotic (electrophoretic) velocity is specified for the case of the complete dissociation of ionic sites within PL. Accordingly, the equation may be used only if the difference between pK and pH is very large. However, it turned out that an analytical solution of the nonlinearized PB equation for thick PL is possible for any degree of dissociation. This was achieved using the approximation of excluded coions if the absolute value of the reduced Donnan potential is larger than 2 and due to the simplification in the case of weak dissociation, when the absolute value of the reduced Donnan potential is less than 2. Combining this generalized double layer (DL) theory for PL and the theory of Ohshima enables to obtain an analytical equation for electroosmosis for the general case of any degree of dissociation. This equation creates for the first time a theoretical base for the interpretation of electrokinetic fingerprinting (EF) for the characterization of soft surfaces. PMID- 15897096 TI - Diffusiophoresis and electrophoresis of a charged sphere parallel to one or two plane walls. AB - The diffusiophoretic and electrophoretic motions of a dielectric spherical particle in an electrolyte solution located between two infinite parallel plane walls are studied theoretically. The imposed electrolyte concentration gradient or electric field is constant and parallel to the two plates, which may be either impermeable to the ions/charges or prescribed with the far-field concentration/potential distribution. The electrical double layer at the particle surface is assumed to be thin relative to the particle radius and to the particle wall gap widths, but the polarization effect of the mobile ions in the diffuse layer is incorporated. The presence of the neighboring walls causes two basic effects on the particle velocity: first, the local electrolyte concentration gradient or electric field on the particle surface is enhanced or reduced by the walls, thereby speeding up or slowing down the particle; second, the walls increase the viscous retardation of the moving particle. To solve the conservative equations, the general solution is constructed from the fundamental solutions in both rectangular and spherical coordinates. The boundary conditions are enforced first at the plane walls by the Fourier transforms and then on the particle surface by a collocation technique. Numerical results for the diffusiophoretic and electrophoretic velocities of the particle relative to those of a particle under identical conditions in an unbounded solution are presented for various values of the relevant parameters including the relative separation distances between the particle and the two plates. For the special case of motions of a spherical particle parallel to a single plate and in the central plane of a slit, the collocation results agree well with the approximate analytical solutions obtained by using a method of reflections. The presence of the lateral walls can reduce or enhance the particle velocity, depending on the properties of the particle-solution system, the relative particle-wall separation distances, and the electrochemical boundary condition at the walls. In general, the boundary effects on diffusiophoresis and electrophoresis are quite significant and complicated, and they no longer vary monotonically with the separation distances for some situations. PMID- 15897097 TI - Force acting on a dielectric particle in a concentration gradient by ionic concentration polarization under an externally applied DC electric field. AB - There is a concentration-polarization (CP) force acting on a particle submerged in an electrolyte solution with a concentration (conductivity) gradient under an externally applied DC electric field. This force originates from the two mechanisms: (i) gradient of electrohydrodynamic pressure around the particle developed by the Coulombic force acting on induced free charges by the concentration polarization, and (ii) dielectric force due to nonuniform electric field induced by the conductivity gradient. A perturbation analysis is performed for the electric field, the concentration field, and the hydrodynamic field, under the assumptions of creeping flow and small concentration gradient. The leading order component of this force acting on a dielectric spherical particle is obtained by integrating the Maxwell and the hydrodynamic stress tensors. The analytical results are validated by comparing the surface pressure and the skin friction to those of a numerical analysis. The CP force is proportional to square of the applied electric field, effective for electrically neutral particles, and always directs towards the region of higher ionic concentration. The magnitude of the CP force is compared to that of the electrophoretic and the conventional dielectrophoretic forces. PMID- 15897098 TI - On the adsorption and diffusion of Methylene Blue in glass fibers. AB - The adsorption and diffusion phenomena of Methylene Blue dye on glass fiber have been explored. Both isotherm and kinetics have been studied. Effect of process parameters such as pH, adsorbent loading, and initial dye concentration was investigated. Temperature had a little effect on adsorption. A diffusion model was developed to estimate the diffusivity of the dye in glass fiber. The diffusivity was found to be varying exponentially with the dye concentration. PMID- 15897099 TI - Synthesis of porphyrin-introduced silica gels by sol-gel process. AB - Using a hydroxyl group appended free base porphyrin derivative (HP), porphyrin introduced silica gels were synthesized by a sol-gel process. The HP content in the materials linearly increased with increasing the HP concentration. Meanwhile, free base tetraphenylporphyrin (TPP) with no hydroxyl groups were almost not incorporated into the silica gels. These facts suggested that the interaction between hydroxyl groups of the HP molecules and silica network is considerably strong. The UV-vis characters of HP-introduced materials were almost the same as pure HP molecules. The Beer's plot indicated that the HP molecules in the materials are dispersed. PMID- 15897100 TI - Photoinduced dissolution and redeposition of Au nanoparticles supported on TiO2. AB - Au particles (mean size ca. 3 nm) supported on TiO(2) particles were irradiated by UV light (>300 nm) in aqueous solutions at 278 K. Photo-induced dissolution of Au nanoparticles followed by redeposition occurred in aqueous solutions containing halogen ions. The dissolution of Au nanoparticles yielded a Au(III) complex with a halogen ion; subsequent reduction of the Au(III) complex caused precipitation of larger Au particles on TiO(2). PMID- 15897101 TI - Evidence-based models for clinical care in pediatric and adolescent gynecology. PMID- 15897102 TI - Lower genital tract disease in children and adolescents--a review. AB - Conditions affecting the lower genital tract of female children and adolescents are often significantly different than those seen in the adult. The purpose of this review is to provide an overview of vulvar, vaginal, and cervical conditions that may be encountered only rarely by the more generalized practitioner. PMID- 15897103 TI - Adolescents' experience with the combined estrogen and progestin transdermal contraceptive method Ortho Evra. AB - BACKGROUND: The new combined estrogen & progestin contraceptive patch Ortho Evra was approved by the FDA in December 2001. To date, there is a paucity of data regarding its use in the adolescent age group. We examined adolescents' experience with this new contraceptive method. METHODS: Using a questionnaire designed by the authors, care providers in a hospital based adolescent clinic interviewed and reviewed the charts of adolescent girls who had initiated Ortho Evra in 2002-2003. RESULTS: Twenty-eight adolescent girls (age 18 +/- 1 years, gyn age 6 +/- 1 years, onset of sexual intercourse at 14 +/- 1 years, body mass index (BMI) 27.6 +/- 1.2, 57% Hispanic, 21% Caucasian, 11% African American, 7% biracial, 4% Indian American) who had used Ortho Evra for 7 +/- 1 months were enrolled. Half (50%) were adolescent mothers, and 57% had a history of irregular menstrual periods. All (100%) girls reported regular menstrual periods while using Ortho Evra, with only 14% experiencing occasional breakthrough bleeding. Half reported a shorter duration and 36% reported a lighter flow of their periods. About a third (39%) reported a decrease and 11% reported an increase in dysmenorrhea symptoms. About a third (29%) of those with a history of recurrent headaches at initiation reported decrease in headaches, and about a third (33%) of those with acne at initiation reported decrease in facial acne while on Ortho Evra. There were no significant BMI changes during Ortho Evra use. Although condom use while on Ortho Evra was poor (only 15% reporting consistent condom use), there were no pregnancies reported. A majority (93%) reported that they remembered to apply the patches on time, and 40% stated that Ortho Evra was easier than previous contraceptive methods. Two thirds (68%) were very satisfied and 29% were somewhat satisfied with the method, and 93% stated that they would recommend the method to a friend/relative. The preferred application site was the buttock (40%) followed by the lower abdomen (32%). About a fifth (21%) experienced at least one episode of complete patch detachment and 32% reported partial peeling of the patch corners. About a third (32%) would prefer another patch color, and 25% would like a fourth week placebo patch. The most common side effects were mild temporary application site reactions (64%), some discomfort on patch removal (32%), nausea (18%), and breast tenderness (18%). Eleven girls (39%) discontinued Ortho Evra (three lost health insurance, three because of application site reactions, two found patch application schedule difficult to remember, two desired pregnancy, two because of nausea, one because of perceived weight gain). CONCLUSIONS: Ortho Evra provides excellent cycle control in adolescents. Most adolescents are satisfied with this method. Intensive efforts should be made to increase condom use by adolescents on Ortho Evra. PMID- 15897104 TI - Sexual victimization: incidence, knowledge and resource use among a population of college women. AB - STUDY OBJECTIVE: To assess the incidence of sexual victimization among a convenience sample of college women and evaluate both victims and non-victims' knowledge and use of available on and off campus resources. DESIGN: Written questionnaire distributed to students in lobbies of two campus libraries and large computing center SETTING: A private northeastern university PARTICIPANTS: Upper-class undergraduate women (sophomores, juniors and seniors) MAIN OUTCOME MEASURES: Respondents (n=234) were asked to complete demographic information, report instances of sexual victimization (including rape, sexual coercion and unwanted sexual contact) that occurred during the 1999-2000 academic year and indicate whether they would or did use either university and/or outside resources available to sexual assault victims. RESULTS: Of the women who participated, 38% (90/234) affirmed one or more episodes of sexual victimization, with 6% (14/234) reporting a completed rape and 4% (9/234) an attempted rape. Drug or alcohol related impairment leading to unwanted sexual activity was reported by 15% (35/234) of women. Utilization of available on- and off-campus resources was uncommon among victims (22% and 6%, respectively); 12% contacted health services, while only 4% reported an event to university security. Victims cited fear, embarrassment and guilt, as well as lack of confidentiality, as the most common reasons for failure to use resources. CONCLUSIONS: Increasing campus awareness of sexual victimization and removing barriers to access for victims should remain university goals. PMID- 15897105 TI - Prenatal diagnostic decision-making in adolescents. AB - STUDY OBJECTIVE: We sought to evaluate the prenatal decision-making of pregnant adolescents identified at increased risk for identifiable fetal genetic abnormalities. DESIGN: A retrospective review of records of gravid women 19 years old or younger undergoing genetic counseling from 2001-2003 (inclusive) was undertaken. SETTING: Hospital-based academic center. PARTICIPANTS: Thirty-seven women were identified; four cases did not meet inclusion criteria. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Decision to undergo or forgo invasive prenatal testing. RESULTS: Of the 33 women included in this study, the average age was 17.6 years (range: 15-19). Eighteen were Latinas, eight were African-Americans, and seven were Caucasians. Sixteen women had positive maternal serum screening outcomes; nine women sought counseling because of personal/family histories of genetic abnormalities, seven sought counseling after fetal structural anomalies were detected by ultrasound, and one woman sought counseling because she and her partner were positive for Mendelian disorder screening (sickle cell disease). Sixteen of the women (48.5%) chose to undergo invasive testing (15 amniocenteses, one chorionic villus sampling) whereas 17 (51.5%) chose to forgo invasive testing. CONCLUSIONS: Adolescents offered invasive prenatal diagnosis will chose to undergo or forgo such testing based on diagnostic and personal criteria as do adult women. Nonetheless, unique adolescent issues may make the process by which information is obtained and communicated during counseling to be different from counseling provided to adults. The development of new genetic screening and diagnostic protocols has and will increase the number of pregnant adolescent women who will be offered genetic counseling during their pregnancies. Such an increase in numbers will place considerably more pressure on an already taxed genetic counseling system; accordingly, new counseling paradigms will need to be developed to provide service to an expanded patient population seeking information for an increasing number of genetic issues. PMID- 15897106 TI - Gynecological care for adolescents with disability: physician comfort, perceived barriers, and potential solutions. AB - OBJECTIVE: The goal of this study was to assess the barriers to gynecologic care for adolescents with disability in this state. DESIGN: Providers of gynecologic care in Utah were surveyed to assess existing conditions, attitudes, and comfort levels regarding this population. RESULTS: The survey was completed by 136 gynecologic clinicians (50% return rate). The comfort level among surveyed clinicians in providing gynecological care to disabled adolescents was relatively low, even for routine screening. Respondents identified the following as barriers to providing this care: time, reimbursement, inadequate knowledge, and disability related issues (office access and preference to not serve this population). Suggested solutions included: continuing education programs, consultation to practices on serving both youth and individuals with disability, and better billing mechanisms. CONCLUSION: While many of the surveyed clinicians are comfortable providing care to adolescents with disability, a substantial number are not. Respondents perceive many barriers to providing care to this population, though the barriers appear modifiable. Potential interventions were identified which should be tested for their effectiveness in improving access to quality care for this underserved population. PMID- 15897107 TI - Reproductive health education and sexual risk among high-risk female adolescents and young adults. AB - STUDY OBJECTIVE: The objective of this study was to explore the associations of sources, content, and timing of reproductive health education with cognitive and behavioral sexual risk in a sample of high-risk female adolescents and young adults. DESIGN: Female adolescents and young adults (n=113, median age 17 years) receiving treatment for a sexually transmitted disease (STD) reported sources of reproductive health education, topics covered, and when first formal education occurred. Dependent variables included sexual risk knowledge; condom attitudes, negotiation skills, and use (consistent and at last sex); and number of sexual partners. RESULTS: Most participants reported receiving reproductive health education from both parental (80%) and formal sources (92%). Parents discussed the menstrual cycle (94%) more frequently than other sex education topics, while formal sources focused most on teaching about STDs (91%). Although median age of first formal instruction was 12 years, 26% of girls received their first formal education during or after the year they initiated coitus. Girls with a parental source of education and those receiving formal instruction on pregnancy reported greater ability to negotiate condom use. Girls who received education later in relation to the onset of sexual activity and those with a parental source of education reported more sexual partners. CONCLUSIONS: Early reproductive health education and education from both parental and formal sources is associated with reduced sexual risk among high-risk adolescent girls. Interestingly, receiving parental education is also associated with more sexual partners, suggesting that parental educational efforts may be reactive to their daughters' increasing sexual risk behavior. Future research should examine multiple sources of reproductive health education and the timing of education from these sources to enhance understanding the dynamic interactions between reproductive health education and adolescent sexual risk. PMID- 15897108 TI - Rare cause of vaginal bleeding in early puberty. AB - STUDY OBJECTIVE: To document an unusual cause of vaginal bleeding in early puberty. DESIGN: Case Report. RESULTS: Clinical presentation with the aid of pelvic ultrasound pointed to the diagnosis of an endometrial polyp in this young adolescent prior to menarche. A hysteroscopy was deemed necessary to allay concerns for a serious pathology and confirm diagnosis. CONCLUSION: Endometrial polyps are infrequent in adolescents but should be included as a differential diagnosis of vaginal bleeding in this population. PMID- 15897109 TI - Beta hemolytic streptococcus group f causing pelvic inflammatory disease in a 14 year-old girl. AB - BACKGROUND: Pelvic inflammatory disease in prepubertal and non-sexually active adolescents is rare and poorly understood. Various organisms have been named as causative agents in adolescent pelvic infections. Early diagnosis and treatment of pelvic inflammatory disease in young girls is imperative to future fertility and long term sequela. CASE: We present a 14-year-old, menarchal, non-sexually active female with a 3-week history of abdominal pain and fever. Surgical exploration and cultures revealed Stage IV pelvic inflammatory disease caused by Beta Streptococcus Group F. CONCLUSION: Various organisms including Streptococcal infections should be considered in the differential diagnosis of pelvic inflammatory disease in young girls without risk factors. PMID- 15897110 TI - What's your diagnosis? Loss of vulvar pigmentation in a two-year-old girl. PMID- 15897111 TI - Uteroovarian ligament torsion of the due to a paratubal cyst. AB - Paratubal cysts rarely undergo torsion. When they do, typically they involve the infundibulopelvic ligament and affect the ovary and fallopian tube. We present a case and discussion of torsion of the uteroovarian ligament secondary to a large paratubal cyst with preservation of the ovary and fallopian tube. PMID- 15897112 TI - Tattoos: beautification or something else? PMID- 15897113 TI - What's new in adolescent contraception: what residents and teachers need to know. PMID- 15897114 TI - Depression and adolescent pregnancy. PMID- 15897115 TI - Chronic administration of angiotensin-(1-7) attenuates pressure-overload left ventricular hypertrophy and fibrosis in rats. AB - OBJECTIVE: To test the hypothesis that chronic administration of angiotensin-(1 7) [Ang-(1-7)] attenuates cardiac hypertrophy in rats in vivo. METHODS: Coarctation of the suprarenal abdominal aorta was performed in 41 8-week-old male Sprague Dawley rats. Twenty-four hours after the operation, osmotic minipumps were surgically implanted subcutaneously in the rats, which were randomly divided into 3 groups, including a sham-operation group (n=15) receiving infusion with normal saline, a suprarenal aortic coarctation group (n=12), and a suprarenal aortic coarctation group (n=14) with Ang-(1-7) treatment at the dose of 25 mug x kg(-1) x h(-1). Four weeks later, the systolic and diastolic blood pressures were measured and the left ventricular mass index (LVMI, mg/g) was calculated from the ratio of left ventricular weight to body weight. The concentrations of Ang II in the plasma and myocardium were measured by radioimmunoassay, and myocardial interstitial collagen volume fraction (ICVF) was determined by quantitative morphometry of the sections with Picrosirius red staining using an automated image analyzer. RESULTS: Suprarenal abdominal aortic coarctation induced a significant increase in carotid artery systolic and diastolic blood pressure, heart weight, LVMI, ICVF, and the concentration of Ang II in the myocardium (P<0.01). Chronic administration of Ang-(1-7) attenuated the increase in the heart weight, LVMI, ICVF and left ventricular diastolic end pressure (LVEDP) caused by suprarenal abdominal aortic coarctation (P<0.05). Ang-(1-7) also increased the formerly decreased maximum left ventricular pressure reduction rate (-dP/dt(max)) (P<0.05), but had no effect on blood pressure and the concentration of Ang II in the myocardium. No difference was noted in plasma concentration of Ang II between the 3 groups. CONCLUSIONS: Ang-(1-7) attenuates cardiac hypertrophy and fibrosis and preserved the impaired left ventricular function induced by left ventricular pressure-overload in rats. These effects are not associated with the changes in the concentrations of Ang II in the left ventricular myocardium and plasma. PMID- 15897116 TI - Effect of one versus two distal locking bolts on the biomechanics of tibial interlocking nail: a comparative study using a new model. AB - OBJECTIVE: To investigate the impact of one versus two distal locking bolts on the mechanical properties of tibial interlocking intramedullary nails. METHODS: Twenty 9-mm titanium alloy locking nails were divided into two equal groups in which the nails were fixed with only one and two distal locking bolts, respectively. Each group was further divided into two sub-groups for compression and torsion tests separately using a universal material-testing machine. RESULTS: In the compression tests, the average maximum strength of double bolts was greater than that of the single bolt (P<0.05), but the number of the bolts did not significantly affect the results of the torsion tests in terms of the either the maximum torsion moment or angle (P>0.05). CONCLUSION: One distal bolt is sufficient for fixing stable fractures and double bolts are recommended for management of serious fractures. The model we used is convenient and economic for examining the biomechanics of the tibial nails, especially for comparative purposes. PMID- 15897117 TI - [Advanced oxidation protein products-induced tumor necrosis factor alpha secretion in monocytes via reactive oxygen species generation]. AB - OBJECTIVE: To investigate the effect of advanced oxidation protein products (AOPP) on the secretion of tumor necrosis factor alpha eTNFalphae in monocytes and its possible mechanism. METHOD: Human monocyte cell line THP-1 and peripheral blood monocytes were incubated with AOPP-bovine serum albumin(BSA) prepared by incubation of BSA with hypochlorous acid. TNFalpha in the supernatant of the culture medium of THP-1 cells was measured by enzyme-linked immunosorbent assay and the production of reactive oxygen species (ROS) evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2,7 dichlorefluorescin using Wallac 1420 multilabel counter. The intracellular signal was observed by pre-treatment of the cells with antioxidant pyrrolidine dithiocarbamate, NADPH oxidase inhibitor apocynin or p38 phosphorylation inhibitor SB203580. RESULTS: AOPP-BSA induced TNF-alpha secretion and ROS production in monocytes. Pretreatment of the cells with pyrrolidine dithiocarbamate scavenged most of ROS and almost completely blocked TNF-alpha secretion induced by AOPP-BSA. Inhibition of NADPH oxidase by apocynin and p38 phosphorylation by SB203580 could both effectively block AOPP-BSA-induced TNF alpha secretion. CONCLUSION: AOPP-BSA induced TNF-alpha secretion in monocytes, and the intracellular signaling involves ROS produced by activated NADPH oxidase and subsequent p38 phosphorylation. PMID- 15897118 TI - [Transplantation of 3H-thymidine-labeled human bone marrow-derived mesenchymal stem cells in mdx mice]. AB - OBJECTIVE: To investigate the feasibility of using human bone marrow-derived mesenchymal stem cells (hBM- MSCs) for repairing the skeletal muscle sarcolemma lesions in mdx mice and characterize the distribution of the transplanted hBM MSCs. METHODS: Eighteen 8- to 10-week-old immunosuppressed mdx mice received transplantation with 1x10(7) of hBM-MSCs (the fifth passage) with 3H-thymidine (3H-TdR) labeling by injection of the cells into the tail vein. The mice were killed at 24 h, 48 h, 2 weeks, and 1, 2 and 4 months after the transplantation, respectively, to measure the radioactivity in the tissues and organs. Dystrophin expression on the sarcolemma was detected by immunofluorescence analysis. RESULTS: One month after transplantation, the mice with cell transplantation showed greater radioactivity in most of the tissues and organs than the control mice, especially in the bone marrow, liver and spleen. The radioactivity was then gradually lowered but in the skeletal muscle, the radioactivity increased progressively since 2 weeks after transplantation, reaching the peak of 27.65+/ 3.53 Bq/mg at 1 month. Compared with that in the control mice, the radioactivity in the bone marrow and skeletal muscle was persistently higher in mice with cell transplantation 1 month after transplantation. No dystrophin-positive cells were found in the mdx mice at 2 weeks but detected at 1 month. The percentage of dystrophin-positive fibers in each section ranged from a 6.6% (1 month) to 8.9% (4 months). CONCLUSIONS: hBM-MSCs engrafted in immunosuppressed mdx mice may differentiate into skeletal muscle cells to repair the pathological lesion of the skeletal muscle sarcolemma. The hBM-MSCs reside mainly in the bone marrow, liver and spleen in the early stage following transplantation, homing into the bone marrow and skeletal muscle later. PMID- 15897119 TI - [Effects of granulocyte colony-stimulating factor on focal cerebral ischemia reperfusion injury in rats]. AB - OBJECTIVE: To study the therapeutic effects of granulocyte colony-stimulating factor (G-CSF) on focal cerebral ischemia-reperfusion injury in rats. METHODS: Adult male SD rats were subjected to transient (2 h) middle cerebral artery occlusion (MCAO) and scored 48 h later for such neurological functions as spontaneous movement, paresis, forelimb motor function, climbing ability, pain sensation, and position sense. Twenty rats with the total score less than 12 were divided equally into two groups to receive daily subcutaneous injection of 20 microg/kg of G-CSF for 19 days or the same dose of saline injection (control group). The rats in the two groups were also given intraperitoneal injection of Brdu10 mg/kg for 19 days. The neurological functions of the rats in both groups were examined on a weekly basis after MCAO and on day 21, the rats were killed to prepare frozen sections of the brain tissues for double immunohistochemical staining with Brdu and glial fibrillary acidic protein antibody to identify the neural cells newly evolved from the stem cells. RESULTS: The rats in G-CSF group showed better neurological function recovery than the control rats 3 weeks after MCAO (P<0.05). Obvious regeneration of the neural cells was observed around the infarction area in the rats receiving G-CSF treatment. CONCLUSION: G-CSF promotes neurological function recovery and neural cell regeneration after cerebral infarction in rats and can be effective for intervention of focal cerebral ischemia-reperfusion injury. PMID- 15897120 TI - [Effect of different tidal volume ventilation on rat bronchial and alveolar epithelial cell apoptosis]. AB - OBJECTIVE: To investigate the changes in bronchial and alveolar epithelial cell apoptosis after mechanical ventilation with different tidal volumes and explore the mechanism and significance of cell apoptosis in ventilation-induced lung injury. METHODS: Thirty healthy male Sprague-Dawley rats were randomly divided into groups A, B and C to receive tracheotomy under anesthesia for ventilation with different tidal volumes (VT) for 4 h (VT of 10 ml/kg in group A, 20 ml/kg in group B and 40 ml/kg in group C). Lung histopathology was assessed after the ventilation and compared between the 3 groups. The severity of lung injury and bronchial epithelial edema was evaluated, and the extent and location of cell apoptosis in the bronchus and lung tissues were studied by transferase d-UTP end labeling (TUNEL) assay and immunohistochemistry. RESULTS: After mechanical ventilation for 4 h, lung injury and bronchial epithelial edema increased with the increment of VT. By TUNEL assay, the apoptosis index of both alveolar and bronchial epithelial cells was found to increase with VT. Immunohistochemical study identified enhanced caspase-3 expression in the cytoplasm and nuclei of airway and alveolar epithelial cells and vascular endothelial cells in rats with high VT ventilation, and the enhancement was especially obvious in the bronchial epithelial cells. CONCLUSION: Mechanical ventilation with different VTs causes consistent changes in histopathology, cell apoptosis and caspase-3 expression in rats, which support the presumption that cell apoptosis plays an important role in the pathogenesis of high VT ventilation-induced lung injury. PMID- 15897121 TI - [Gene reversion of induced differentiation of adult human bone marrow-derived mesenchymal stem cells into neuron-like cells]. AB - OBJECTIVE: To observe gene reversion during differentiation of human adult bone marrow-derived mesenchymal stem cells (MSCs) into neuron-like cells induced by Shenqiye. METHODS: The MSCs were separated, cultured and expanded in the culture medium and induced to differentiate into neuron-like cells with Shenqiye. The expressions of neuron-specific enolase (NSE), neurofilament (NF), and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method, and the changes of 10 genes of the cells after differentiation were detected by reverse transcriptional PCR. RESULTS: The MSCs exhibited neuronal phenotype when treated with Shenqiye and the neuron-like cells were positive for expressions of NSE and NF but not for glial astrocyte marker GFAP. After withdrawal of Shenqiye from the medium, the neuron-like cells were reversed to MSC, flat or spindal in morphology. The gene expression profiles of the redifferentiated cells were similar to those of undifferentiated MSCs. CONCLUSION: Shenqiye can induce MSC differentiation into neuron-like cells during and after which gene reversion may occur. PMID- 15897122 TI - [Effect of KDR recombinant adenovirus containing double suicide gene on proliferation, apoptosis and cell cycle of human umbilical vein endothelial ECV304 cells]. AB - OBJECTIVE: To study the effect of adenovirus (Ad)-mediated fusion gene systemdriven by KDR promoter on the proliferation, apoptosis and cell cycle of human umbilical vein endothelial ECV304 cells. METHODS: The KDR-expressing ECV304 cells and LS174T cells not expressing KDR were both infected by the AdEasy-KDR CDglyTK followed by treatment with the prodrugs 5-flurocytosine (5-FC) and/or ganciclovir (GCV) at different concentrations. The killing effects of the transfection on the cells were evaluated and bystander effects analyzed by coculturing the uninfected cells by AdKDR-CDglyTK with different ratios of infected cells. Flow cytometry was employed for determining the cell cycle distribution and electron microscopy performed to observe the pathological changes of cells. RESULTS: The infection rates of the resultant recombinant Ad (rAd) were similar in the cells and gradually increased with the increment in the multiplicity of infection (MOI) of the Ads. The infected cells exhibited different sensitivities to the two prodrugs: ECV304 cells infected with rAd were highly sensitive to the prodrugs, but the infected LS174T cells were not (P<0.001). The killing effect of CD/TK fusion gene on the target cells was much stronger than that of either single suicide gene (P<0.001), showing also obvious bystander effect. In addition, the cell cycle of ECV304 cells was arrested at S phase with morphologic features of apoptosis and necrosis as displayed by electron microscopy. CONCLUSIONS: CD/TK fusion gene system driven by KDR promoter selectively kills the KDR-CDglyTK-expressing endothelial cells, the mechanism of which may involve cell cycle arrest and necrosis and apoptosis of the cells. PMID- 15897123 TI - [In vitro antitumor effect of adenovirus containing CD-TK fusion gene driven by VEGF promoter on LoVo cells]. AB - OBJECTIVE: To evaluate the in vitro antitumor effect of adenovirus containing cytosine deaminase (CD) and thymine kinase (TK) gene driven by vascular endothelial growth factor (VEGF) promoter (Ad-VEGFp-CDglyTK) on human colorectal cancer LoVo cells. METHODS: The adenoviral vectors Ad-VEGFp-CDglyTK, Ad-CMV-CD, Ad-CMV-TK and Ad-CMV- CDglyTK were transfected into LoVo cells, which were then treated with the prodrugs 5-FC or GCV to observe the plating efficiency and growth inhibition effect and bystander effect of the vectors on LoVo cells. RESULTS: Compared with the two vectors containing single suicide gene, Ad-VEGFp CDglyTK showed more remarkable inhibitory effect on the plating efficiency and growth of the tumor cells, and the bystander effect of the fusion suicide gene was much stronger than that produced by single suicide gene. But the LoVo cells infected with Ad-VEGFp-CDglyTK exhibited similar sensitivity to the prodrugs with the cells infected with Ad-CMV-CDglyTK. CONCLUSION: Ad-VEGFp-CDglyTK has potent antitumor effect on LoVo cells in vitro, which is comparable with the effect of Ad-CMV-CDglyTK. PMID- 15897124 TI - [Adenovirus-mediated CDglyTK fusion gene system driven by KDR promoter selectively kills colorectal cancer cells]. AB - OBJECTIVE: To observe the selective killing effect of adenovirus (Ad)-mediated double suicide gene driven by kinase domain-containing receptor(KDR) promoter on human colorectal cancer LoVo cells and human umbilical vein endothelial ECV304 cells. METHODS: The plasmid pAdEasy-KDR-CDglyTK was transfected into 293 packaging cells for amplification of the infectious Ad and used to infect the KDR producing cells (ECV304 and LoVo) and the KDR-nonproducing cells (LS174T) respectively. The three cells were treated with the prodrugs 5-flurocytosine (5 FC) and ganciclovir (GCV) at different concentrations after infection. The killing effects of the fusion gene system on the cells were evaluated. The distribution of cell cycle was detected by flow cytometry. RESULTS: The infection rates of the recombinant Ad were similar among the 3 cells, gradually increasing with the increment of multiplicity of infection (MOI) and reaching 100% with the MOI of 200. The LoVo cells and ECV304 cells infected with Ad-KDR-CDglyTK were highly sensitive to both of the prodrugs (P>0.1), whereas the infected LS174T cells failed to exhibit similar sensitivity (P<0.001). The killing effect of CD/TK fusion gene on the target cells was much stronger than that of either suicide gene (P<0.001). The cell cycle of LoVo cells was arrested at G1 phase. CONCLUSION: The CD/TK fusion gene system driven by KDR promoter can selectively kill KDR-expressing human colorectal cancer LoVo cells and endothelial cells. PMID- 15897125 TI - [Soluble expression and characterization of the immunoreactive multiepitope antigen of Toxoplasma gondii in E.coli]. AB - OBJECTIVE: To obtain soluble expression product of immunoreactive recombinant multiepitope antigen of Toxoplasma gondii from E.coli. METHODS: The gene encoding the multiple epitopes (MEG) of Toxoplasma gondii was amplified by PCR from the original plasmid containing MEG gene and cloned into the prokaryotic soluble expression vector pET32a. After identification by enzyme digestion and sequencing, the positive recombinant plasmid pET32a-MEG was transformed into BL21(DE3), which was induced with IPTG for expression of the target antigen. The relative molecular mass, solubility and antigenicity of the expression products were analyzed by SDS-PAGE and Western blotting. RESULTS: The recombinant expression plasmid pET32a-MEG was successfully constructed and the highly efficient expression of the antigen was achieved after IPTG induction of E.coli. Improvement of the induction condition increased the expression product which accounted for about 28% of the total bacterial protein. The target protein, with good solubility and a relative molecular mass of about 31 000, was purified by immobilized metal affinity chromatography (Ni-NTA resin) and could be well recognized by mouse and rabbit antisera derived by infection of the animals with Toxoplasma gondii B36 and RH, respectively. CONCLUSION: The recombinant multiepitope antigen has good antigenicity and potential value in diagnosis and vaccine development of toxoplasmosis. PMID- 15897126 TI - [Preparation oral liposome-encapsulated recombinant Helicobacter pylori heat shock protein 60 vaccine for prevention of Hp infection]. AB - OBJECTIVE: To prepare oral liposome-encapsulated recombinant Helicobacter pylori (Hp) heat shock protein 60 (Hsp60) vaccine and investigate its effect against Hp infection in mice. METHODS: The recombinant vector PET-22(+)/Hsp60 was transformed into BL21(DE3) E.coli. The recombinant protein was purified with Ni NTA agrose resin and the oral liposome-encapsulated vaccine was prepared with phosphatidyl choline and cholesterols using film method, with the size distribution of the folate liposomes measured by transmission electronic microscopy. BALB/c mice were divided into 5 groups and immunized by intragastric administration of PBS, liposome, rHsp60 plus choleratoxin (CT), liposome encapsulated rHsp60, and liposome-encapsulated rHsp60 plus CT, respectively, given once a week for 4 weeks. All the mice were challenged by Hp for 3 times within two weeks following the last immunization and sacrificed 3 weeks after the last challenge. Hp detection was performed by fast urease test. Semi-quantitative assessment of the bacterial colonization density observation of the inflammation severity and gastric histopathology were carried out. RESULTS: The soluble expression product accounted for 27% of the total bacterial protein. The purity of recombinant fusion protein was about 95% after purification. The mean size of the folate liposomes was 0.7+/-0.4 mum. PBS or liposome alone showed no immune enhancing effect, and rHsp60 plus CT, liposome-encapsulated rHsp60 and liposome encapsulated rHsp60 plus CT had the protective rates against Hp infection of 73.3%, 66.7% and 86.7%, respectively. The latter 3 preparations effected significantly reduced Hp infection and alleviated the inflammation in the gastric mucosa of the mice challenged with Hp. CONCLUSION: The oral liposome may serve as a potential adjuvant for Hp vaccine in preventing Hp infection. PMID- 15897127 TI - [In vitro culture of multipotent adult progenitor cells purified from human bone marrow by magnetic-activated cell sorting]. AB - OBJECTIVE: To establish a method for purifying and culturing human multipotent adult progenitor cells (MAPCs) in vitro. METHODS: Mononuclear cells were separated from the bone marrow of healthy adult volunteers by density gradient centrifugation and cultivated in adherent culture. The plastic-adherent cultured bone marrow cells were isolated by magnetic-activated cell sorting with CD45 and GlyA magnetic microbeads, and the purity of CD45-/GlyA- cells evaluated by flow cytometry. Phase-contrast microscopy was used to detect morphological changes of the cells in different stages of culture. RESULTS: Approximately (5-10)x10(4)/ml MAPCs could be separated from every 1x10(6)/ml bone marrow mononuclear cells by magnetic- activated cell sorting. The viability of the cells before and after separation was (96.7+/-1.7)% and (96.0+/-2.4)%, respectively. The isolated MAPCs grew well in a self-prepared culture medium till the16th passage. The purity of CD45-/GlyA- separated from the bone marrow was more than 98% as examined by flow cytomety even till the 12th passage. CONCLUSIONS: MAPCs derived from adult human bone marrow can be purified by magnetic-activated cell sorting with CD45 and GlyA microbeads and retain the undifferentiated state for a long time. The self prepared culture medium is appropriate for MAPCs cultures in vitro. PMID- 15897128 TI - [A colloidal gold immunochromatographic assay for detecting p38 antigen of Schistosome japonicum]. AB - OBJECTIVE: To establish a colloidal gold immunochromatographic assay (GICA) for detecting Schistosome japonicum (Sj). METHODS: Eight monoclonal antibodies (mAbs) against Sj p38 antigen (1A6, 3C4, 3D12, 6F10, 6G12, 9H6, 9G7 and A5H) prepared previously were purified by protein-G affinity chromatography. The affinity constant (K(aff)) was determined by indirect enzyme-linked immunosorbent assay. All the mAbs were labeled with horseradish peroxidase by sodium oxidation method and the mAb pairs with high affinity and stability were identified according to their optical density at 450 nm (OD450). Four mAbs (1A6, 6G12, 9H6 and 9G7) were chosen for colloidal gold labeling. GICA was then performed by further optimization of the labeling and the conditions were determined. The sera of mice at different infection stages were examined with GICA dipstick, with the sera collected before infection and those of Toxoplasma gondii-infected mice as negative controls. RESULTS: The purity of the 8 mAbs was higher than 95% with K(aff) ranging from 2.8x10(-10) to 1x10(-8) mol/L. 9G7 coating (2.5 mg/ml) as the capture antibody and detection with 1A6 (diluted at 1:4) as the labeling antibody was determined as the best reaction model. With this combination, the positivity rates of the detection were 40%, 50%, 60% and 80% for mouse sera collected at 3, 4, 5 and 6 weeks after Schistosome japonicum infection, respectively, without positive results for the negative control samples. CONCLUSION: GICA established in this study is characterized by simplicity, rapidity and good sensitivity, and the prepared rSjP38 dipstick can test the circulating antigen SjP38 in early stage of infection. PMID- 15897129 TI - [Photon transport in pelvic radiotherapy for cervical cancer]. AB - OBJECTIVE: At present clinical treatment of cervical cancer combines surgical intervention, radiotherapy and chemotherapy. Optimal doses in radiotherapy help improve survival of the patients, but the decision on such dose has to originate from direct measurement. Due to the complexity in the anatomic structure of female pelvic cavity and the adjacent tissues, quite an array of measurement equipments, experimental techniques and radiation parameters have to be employed or determined during radiation measurements, which adds to the complexity of measurement as well as the costs. In this article, the author introduces random photon tracing simulation method to compute tissue photon flux during pelvic radiotherapy for cervical cancer. Besides the simplicity, flexibility and accuracy, this method also lessens the influences by the conditions of and is well adapted to complicated structure of the female pelvic cavity, drastic changes in tissue parameters, and anisotropic scattering. PMID- 15897130 TI - [High-contrast resolution measurement of CT and phantom testing]. AB - OBJECTIVE: The authors adopted subjective and objective methods for measuring high-contrast resolution (HCR) of CT and explored the parameters in relation to HCR. With Catphan500 and a self-prepared experimental phantom, the two CT sets were tested for HCR with varied slice thickness and kernels. The subjective assessment of the images was carried out by visual detection of HCR, and objective assessment was performed by calculations. Each of the methods has its advantages and limitations, and many factors may affect the HCR of CT. PMID- 15897131 TI - [Therapeutic effects of koumine on psoriasis: an experimental study in mice]. AB - OBJECTIVE: To study the therapeutic effects of koumine on psoriasis in mouse models. METHODS: The effects of koumine on epithelial cell mitosis and epidermal cell differentiation was evaluated by collecting the samples of the vaginal mucous and squamous epidermis at the tail of mice treated with methotrexate or koumine at different doses. The levels of interleukin (IL)-2 were detected with enzyme-linked immunosorbent assay. RESULTS: High and intermediate doses of koumine showed remarkable inhibitory effect on mouse vaginal epithelial cell mitosis and promoted the formation of epidermal granular layer in the scales at the mouse tail. Three concentrations at 6, 30, 150 mg/kg of koumine decreased serum IL-2 level in the mice. CONCLUSION: The therapeutic effect of koumine against psoriasis is related to the inhibition of epidermal cell proliferation, promoting the formation of granular cells and decreasing the serum level of IL-2. PMID- 15897132 TI - [Quantitative analysis of serum troponin I and myoglobin in patients with acute myocardial infarction]. AB - OBJECTIVE: To investigate the clinical implications of cardiac troponin I (cTnI), myoglobin (Mb), creatine kinase (CK) and creatine kinase isoenzyme (CK-MB) in patients with acute myocardial infarction (AMI). METHODS: The serum concentrations of cTnI and Mb were determined in 50 patients with AMI 2 to 4 h after chest pain onset and compared with those measured in 50 age-matched healthy subjects. RESULTS: The serum levels of cTnI, CK-MB and CK in patients with AMI were significantly higher than those in the control group. Mb levels in AMI group were above the normal range. Mb, CK-MB, CK and cTnI reached the peak levels at 8+/-2.2, 18.1+/-3.2, 19.4+/-4.1 and 18.6+/-2.9 h after the onset, respectively, and their levels increased with the aggravation of AMI and were reduced to the normal levels with the amelioration of the disease. All the 4 indices were normal in the control group. CONCLUSION: cTnI and Mb are reliable biochemical markers for early diagnosis of AMI and the changes in their serum levels have clinical significance in diagnoses and prognostic judgment of AMI. PMID- 15897133 TI - [Self-efficacy and health behaviors associated with osteoporosis of middle-aged and elderly subjects: a community-based study]. AB - OBJECTIVE: To understand the self-efficacy and health behaviors with respect to osteoporosis of middle-aged and elderly subjects living in the communities. METHOD: A questionnaire-based survey was conducted using the Osteoporosis Self Efficacy Scale (designed by Horan M et al ) and osteoporosis health behaviors questionnaire designed by the authors among 290 middle-aged and elderly community dwellers. RESULTS: The mean score of the self-efficacy was 64.75+/-21.93 among the involved subjects without statistically significant difference in terms of gender and intensity of daily exercises. The score differed significantly, however, in terms of age, education background, occupation and calcium intake (P<0.01), and the older group had lower scores than the younger one. Subjects with higher education scored higher, and those with occupations of physical labor scored lower than those with intellectual work. Higher calcium intake also resulted in higher score. It was found in this survey that the preventive behaviors of osteoporosis was insufficient among the subjects, and only 36.1% of the subjects drank milk everyday with 29.2% often having bean products and 49.8% taking calcium supplements. A percentage of 58.1% of these subjects regularly took part in exercises more than 3 times a week for over 30 minutes each time, but they seldom took weight bearing exercise. CONCLUSION: Osteoporosis self efficacy should enhanced to promote osteoporosis- preventing behaviors in osteoporosis education program for the middle-aged and elderly people in the communities. PMID- 15897134 TI - [A questionnaire-based survey of the quality of life of 783 air-force soldiers and officers in the southern military region of China]. AB - OBJECTIVE: To investigate the quality of life of the air-force soldiers and officers in the southern military region of China. METHODS: A self-rated health measurement scale was used to evaluate the quality of life of 783 air-force soldiers and officers in view of their physical, mental and social statuses. RESULTS: The quality of life of the subjects was at an intermediate level. Their physical, mental and social scores were 66.33+/-9.38, 58.99+/-10.09 and 53.69+/ 12.40, respectively, with a total score of 178.44+/-22.60. The quality of life of the soldiers and officers significantly differed with educational background (P<0.01), places of enlisting (P<0.01), arm of the service (P<0.05), identities or positions (P<0.01), and occupation before enlisting (P<0.01). CONCLUSIONS: The quality of life of the soldiers and officers can be affected by such factors as educational back- ground, places of enlisting, arm of the service, identities or positions, and occupation before enlisting, but the nationalities or status of being the only child in family does not obviously influence the quality of life of the military personnel. PMID- 15897135 TI - [Ultrasonographic diagnosis of central slip rupture in the finger extensor mechanism of the proximal interphalangeal joint]. AB - OBJECTIVE: To evaluate the accuracy of ultrasonographic diagnosis for central slip rupture of the finger extensor tendon mechanism of the proximal interphalangeal joint. METHODS: Sixty-seven consecutive patients with injuries to the proximal interphalangeal joint underwent ultrasonic examination with a linear 5-10 MHz hockey stick-shaped scanner head. Meanwhile three methods of physical examination were applied to assess the injury, and the results of ultrasonic detection and physical examinations were compared with surgical findings. RESULTS: Thirty-nine patients with central slip rupture of the finger extensor mechanism were confirmed. Ultrasonography achieved good consistence with surgical findings in injury detection. For ultrasound and physical examinations, the sensitivity for injury detection was 100% and 89.74% with specificity of 96.43% and 82.14%, respectively, showing significant difference (P<0.01). CONCLUSION: As an accurate noninvasive method, ultrasonographic diagnosis can identify central slip rupture in the extensor mechanism of the finger over the proximal interphalangeal joint in early stages. PMID- 15897136 TI - [Prokaryotic expression vector construction, expression and polyclonal antibody preparation of the fusion protein of glutathione S-transferase and peroxisome proliferator-activated receptor-gamma coactivator-1]. AB - OBJECTIVE: To express the fusion protein of glutathione S-transferase (GST) and peroxisome proliferator-activated receptor-gamma coactivator-1 (PPARgammaC1) in E. coli. and prepare the polyclonal antibody against PPARgammaC1. METHODS: The coding sequence of PPARgammaC1 gene was amplified by reverse transcriptase-PCR (RT-PCR) from the total RNA of Hep G2 cells and inserted into pGEX-4T-1 vector. The recombinant vector was identified by restriction endonuclease digestion analysis and the fusion protein GST-PPARgammaC1 was expressed in E. coli. via IPTG induction. The expressed fusion protein was purified by glutathione-agarose affinity chromatography and used to immunize the egg-laying hens for preparing the polyclonal antibody against GST-PPARgammaC1. RESULTS: Restriction endonuclease digestion analysis demonstrated that the PPARgammaC1 gene had been correctly inserted into pGEX-4T-1 vector, and the expressed fusion protein had a relative molecular mass of approximately 39,000 as shown by SDS-PAGE. The polyclonal antibody obtained from the egg yolk immunoglobulins was found to specifically bind to purified PPARgammaC1 in Western blot analysis. CONCLUSION: The successfully prepared polyclonal antibody against PPARgammaC1 peptide provides a useful reagent for PPARgammaC1 detection. PMID- 15897137 TI - [Effect of koumine on proliferation of murine CD4+ T cells purified by magnetic activated cell sorting in vitro]. AB - OBJECTIVE: To assess the separation efficiency of magnetic-activated cell sorting in the purification of CD4+ T cells from murine spleen, and observe the effects of koumine on the proliferation of the separated cells. METHODS: CD4+ T cells were isolated from murine spleen by magnetic-activated cell sorting (MiniMACS). Fluorescence-activated cell sortering was employed to determine the purity of CD4+ T cells before and after the separation procedure followed by evaluation of the cell viability using trypan blue staining. Concanavalin A- (ConA, 5 microg/ml) or phytahematoagglutinin (PHA,1 mg/ml)-induced murine T cells were treated with different concentrations of koumine (10-320 microg/ml), and their proliferation was determined by MTT colorimetry, and enzyme-linked immunosorbent assay was used to measure IL-2 level in the cell culture supernatant. RESULTS: The purity of CD4+ T cells reached (90.3+/-5.8)% after the purification with a cell viability of (94.9+/-3.6)%. Koumine (20-320 microg/ml) dose-dependently inhibited ConA- or PHA-induced proliferation of murine lymphocytes as compared with the controls (P<0.05). Koumine (20, 100, and 200 microg/ml) significantly decreased the level of IL-2 in comparison with the control group (P<0.05). CONCLUSIONS: CD4+ T cells of high purity can be obtained from murine spleen using MiniMACS without impairing the viability of the cells. Koumine significantly inhibits the proliferation of murine CD4+ T cells due to its immunosuppressive effect and inhibition of IL-2 secretion. PMID- 15897138 TI - [Acute toxicity of microcystin-LR in BALB/c mice]. AB - OBJECTIVE: To investigate acute toxicity of microcystin-LR in BALB/c mice. METHODS: BALB/c mice were subjected to intraperitoneal injection of microcystin LR at the half lethal dose (LD50) and 1/2 LD50, and the organ weight indices and various biochemical indices were determined 30 min and 4 h after the treatment, respectively. RESULTS: Significant increase in liver or kidney weight index occurred in the treated mice, indicating hemorrhagic conditions of the two organs. A slight increase in serum levels of aspartate amino transferase and alanine aminotransferase was observed along with an obvious decrease in serum urea levels, suggesting liver damage. The increase of serum uric acid also indicated the presence of kidney damage. CONCLUSION: The kidney may be another target of microcystins besides the liver. PMID- 15897139 TI - [Expressions of transforming growth factor-beta1 and collagen IV in renal tissues of patients with chronic allograft nephropathy]. AB - OBJECTIVE: To investigate the expressions of transforming growth factor (TGF) beta1 and collagen IV in the renal tissues of patients with chronic allograft nephropathy (CAN). METHODS: Immunohistochemical method and computer-assisted image analysis system were used to detect the expressions of TGF-beta1 and collagen IV in the renal tissues of patients with CAN, and the association between TGF-beta1 and collagen IV expressions as well as that between their expressions and the pathological grading of CAN were analyzed. RESULTS: The expressions of TGF-beta1 and collagen IV were significantly higher in the renal tissues of the patients than in normal renal tissues (P<0.001), and the expressions tended to increase with the pathological grades of CAN; TGF-beta1 and collagen IV expressions in both the renal glomeruli and the tubulointerstitium were in patients with CAN positively correlated with normal renal tissues (r=0.943, P<0.001; r=0.910, P<0.001). CONCLUSIONS: Abnormal collagen IV deposition is one of the major factors associated with renal fibrosis in CAN, and TGF-beta1 might play an important role in renal fibrosis in CAN through up regulation of collagen IV in the renal tissues. PMID- 15897140 TI - [Predictive value of endometrial ultrasonography and age for the outcome of in vitro fertilization-embryo transfer]. AB - OBJECTIVE: To assess the predictive value of endometrial ultrasonography and age for the outcome of in vitro fertilization-embryo transfer (IVF-ET). METHODS: A total of 305 women with tubal occlusion were studied. Superovulation was induced and transvaginal ultrasonography was employed for monitoring the endometrial thickness and endometrial patterns on the day of human chorionic gonadotrophin (hCG) administration. Pregnancy was diagnosed by ultrasonography of the intrauterine gestational sac. RESULTS: No difference was noted in the endometrial thickness or endometrial patterns between pregnant and nonpregnant groups, but the least endometrial thickness essential for pregnancy was 7 mm. The pregnant rate of women aged 20-34 years was higher than that of women aged 35 years or above. CONCLUSIONS: Neither endometrial thickness nor endometrial patterns detected by transvaginal ultrasonography on the day of hCG administration allows a reliable prediction of pregnancy. The least endometrial thickness essential for pregnancy is 7 mm and the pregnant rate of women aged 20-34 years was higher than those aged 35 years and above. PMID- 15897141 TI - [Clinical study of lymph node metastatic pattern of thoracic esophageal squamous carcinoma]. AB - OBJECTIVE: To analyze the patterns of lymph node metastases of thoracic esophageal carcinoma and define the adequate range of lymph node dissection. METHODS: The clinical data of 217 patients with esophageal carcinoma undergoing radical surgical resection of the lymph nodes in three regions were retrospectively analyzed. RESULTS: Lymph node metastases were found in 136 of the 217 patients (62.6%) and skip metastases of the lymph nodes in 12 patients (5.5%). In 3 989 lymph nodes desected, metastases were identified in 454 lymph nodes (11.38%). The rates of lymph node metastasis were 31.7%, 21.2% and 12.1% in the neck, thoracic mediastinum and abdominal cavity, respectively, in upper thoracic esophageal carcinoma, 21.9%, 30.5% and 15.6% in middle thoracic carcinoma, and 9.75%, 12.7% and 34.5% in lower thoracic carcinoma. The degree of tumor differentiation, depth of tumor invasion and lymphatic vessel invasion were factors influencing lymph node metastases (P<0.05). CONCLUSION: Because of the upward, downward and skip metastasis of esophageal carcinoma cells to the lymph nodes, the operable patients with thoracic esophagus carcinoma should receive radical desection of the lymph nodes in the 3 regions to promote the patients' survival. PMID- 15897142 TI - [Factors associated with acute ST-segment elevation myocardial infarction with normal angiographic findings of the coronary artery]. AB - OBJECTIVE: To identify the factors related to acute ST-segment elevation myocardial infarction (STEMI) with normal angiographic findings of the coronary artery. METHODS: An retrospective analysis of the electrocardiographic, echocardiographic, and angiographic data of 271 STEMI cases was conducted. Of these patients, 29 had normal coronary artery by angiography and from the rest patients presenting abnormal angiographic findings of the coronary artery, 60 were randomly selected to serve as the control group. Multiple logistic regression analysis was performed to identify the independent factors related to acute STEMI with normal coronary artery by angiography. RESULTS: The incidence rate of STEMI with normal coronary artery was 10.7%. Univariate analysis showed that age, smoking, diabetes mellitus, absence of pre-infarction angina, and wall motion score were related to STEMI with normal coronary artery (P<0.05), whereas multiple logistic regression analysis identified the former 3 factors as the related factors (P<0.05). Wall motion score, left ventricular ejection fraction, cardiac index, and stroke volume index were higher, and cardiac events fewer in patients with normal coronary artery than in those with abnormal coronary artery (P<0.01). CONCLUSION: Acute STEMI with normal coronary artery is more likely to occur in young smokers without pre-infarction angina, possibly in association with spontaneous reperfusion. PMID- 15897143 TI - [Protective effects of losartan against myocardial ischemic reperfusion in intact canine]. AB - OBJECTIVE: To evaluate the protective effect of losartan, an angiotensin II type 1 (AT1) receptor antagonist, against myocardial ischemic reperfusion in intact canine. METHODS: Sixteen dogs were divided into model group(n=8) with left anterior descending coronary artery occlusion for 30 min followed by reperfusion for 30 min and losartan group (n=8) with 5 mg/(kg x d) intragastric losartan administration for 2 weeks before the ischemic-reperfusion model establishment. Monophasic action potential recording technique was employed to observe the incidence of early after depolarization (EAD) and reperfusion arrhythmias. RESULTS: In the model group, 5 dogs (62.5%) developed EAD and reperfusion arrhythmia while 3 (37.5%) developed the symptoms in losartan group, showing significant difference between the two groups (P<0.05). No ventricular tachycardia or ventricular fibrillation was observed in losartan group (P<0.01). CONCLUSION: Losartan can suppress the occurrence of EAD during reperfusion following myocardial ischemia to reduce the incidence of reperfusion arrhythmias, suggesting its protective effect against myocardial ischemic injury. PMID- 15897144 TI - [Effect of terbinafine in treating onychomycosis in children: clinical observation of 88 cases]. AB - OBJECTIVE: To observe the therapeutic effect and safety of terbinafine in treating onychomycosis in children. METHODS: Terbinafine was used to treat 50 children with fingernail onychomycosis and 38 with toenail onychomycosis, and the efficacy, fungal clearance and safety of the drug were evaluated. RESULTS: The clinical cure rate and total effective rate were 92.1% and 97.37%, respectively, for fingernail onychomycosis, and 86.36% and 93.94% for toenail onychomycosis, with fungal clearance rate of 96.59%. No serious side-effects was observed in the children receiving terbinafine therapy. CONCLUSION: Terbinafine is effective and safe for the treatment of onychomycosis in children. PMID- 15897145 TI - [Expression and significance of fragile histidine triad in oral cancer and precancerous lesions]. AB - OBJECTIVE: To detect the expression of fragile histidine triad (FHIT) in oral cancer and oral precancerous lesions and investigate the relationship between the FHIT expression and the histopathological changes. METHODS: Immunohistochemical staining by SP methods was utilized to detect the expression of FHIT in 64 cases of oral squamous cell carcinoma, 39 oral precancerous lesions and 12 normal oral mucosa specimens. RESULTS: The positivity rate of FHIT in normal oral membrane was 100% (12/12), and medium and high FHIT expression levels were detected in oral precancerous lesions but without significant difference from that in normal oral mucosa. In 64 oral squamous cell carcinoma specimens, 3 were negative for FHIT expression, 8 had low expression, and the other 43 had moderate and high expression. The rate of negative or low FHIT expression in the oral squamous cell carcinoma was 17% (11/64), which was significantly different from that in the normal oral mucosa and oral precancerous lesions, but the rate was not correlated to the differentiation of the cancer. CONCLUSION: The tumor suppressor gene FHIT plays a role in the pathogenesis of oral squamous cell carcinoma. PMID- 15897146 TI - [Emergency bedside temporary cardiac pacing through the left subclavicular vein guided by pacemaker impulse]. AB - OBJECTIVE: To investigate the feasibility and merits of emergency bedside temporary cardiac pacing through left subclavicular vein guided by pacemaker impulse. METHODS: The left subclavicular vein was punctured with Seldinger technique for insertion of a cannula. After setting the rate of pacemaker at 20 beat/min above the patients' spontaneous heart rate with output current of 5 mA and sensing voltage of 3 mV, the electrode was inserted through the cannula until the electrocardiograph displaying pulsed signal and pacing rhythm. RESULTS: Good pacing was achieved in 18 patients with one having poor pacing, which was improved 2 h later. The pacing was not satisfactory in two cases. No postoperative complications occurred in the 21 patients. CONCLUSION: Emergency bedside temporary cardiac pacing through the left subclavicular vein guided by pacemaker impulse is rapid and ensures high success rate and safety. PMID- 15897147 TI - [Diagnosis and treatment of traumatic splenic rupture in children: report of 70 cases]. AB - OBJECTIVE: To evaluate the choice of treatment method for traumatic splenic rupture in children. METHODS: A retrospective analysis of 70 children with traumatic splenic rupture was performed. RESULTS: Of the 70 children, 10 were managed with non-surgical treatment and cured. In the 60 children receiving operations, 35 underwent partial splenectomy or prosthesis, 25 had total splenectomy, and the other 21 received autologous spleen transplantation. The follow-up study of 61 children lasting for 3 months to 5 years after operation did not reveal the occurrence of overwlrelmingpostsplenectomy infection. CONCLUSION: The decision on adequate management of traumatic splenic rupture in children should be made on the basis of the patient's condition and severity of spleen injury, and autologous spleen transplantation can be safe and effective in children with splenectomy. PMID- 15897148 TI - [Diagnosis of simple endobronchial tuberculosis by fiberobronchoscopy]. AB - OBJECTIVE: To assess the efficacy of fibrobronchoscopy in diagnosis of simple endobronchial tuberculosis. METHOD: Biopsy, brush biopsy and bronchoalvelar lavage fluid examination were performed in 92 patients with simple endobronchial tuberculosis using an electronic fibrobronchoscope. RESULTS: Simple endobronchial tuberculosis commonly occurred in young people (25-35 years of age), often developed in the superior lobe apicoposterior segment, middle lobe (ligule) and superior branch of the inferior lobe of the left lung. Under fibrobronchoscope, congested edema, caseous necrosis and ulcer granulation were observed in patients with simple endobronchial tuberculosis. The successful detection rates for biopsy, brush biopsy and bronchoalvelar lavage liquid examination were 81.5%, 30.4% and 20.7%, respectively. CONCLUSION: Fiberobronchoscopy is effective for diagnosis of simple endobronchial tuberculosis. PMID- 15897149 TI - [Clinical observation of liver damages in epidemic hemorrhagic fever]. AB - OBJECTIVE: To study the changes in liver function in patients with epidemic hemorrhagic fever (EHF). METHOD: A retrospective analysis of the laboratory tests of liver function in 103 patients with EHF and control subjects was carried out. RESULTS: The alanine aminotransferase (ALT), serum bilirubin (SB) and albumin levels in patients with severe and critical liver damages significantly differed from those in milder cases, and more sever liver damages were found in the death cases of EHF than in survived patients in terms of ALT and SB increment (P=0.000). The liver damage was related to the clinical type, age and the levels of blood platelet , albumin, lactic dehydrogenase and creatinine in EHF cases (P=0.000, 0.000, 0.030, 0.010, 0.000 and 0.013, respectively). CONCLUSIONS: Liver damage is common in patients with EHF and exacerbates with the severity of the disease. Monitoring of ALT, SB and albumin is helpful for evaluating the progression and prognosis of EHF. PMID- 15897150 TI - [Effect of high salt diet on expression of osteopontin mRNA in Sprague-Dawley rat kidney]. AB - OBJECTIVE: To investigate the effect of high-salt diet on the expression of osteopontin (OPN) mRNA and its protein in Sprague-Dawley rat kidney. METHODS: Forty-eight male SD rats aged 10 weeks receiving normal salt diet were enrolled in study, and were divided into two groups and fed with high salt diet (4%NaCl) or normal salt diet (0.6%NaCl) for 12 weeks respectively, with 24 rats in each group. Tail systolic blood pressure and bodyweight were measured in all rats every week. At the end of 4th, 8th and 12th week, 6 rats in each group were sacrificed for detection of the expression of OPN mRNA and its protein in the kidney with quantitatine real-time (QRT)-PCR and immunohistochemistry, respectively. RESULTS: No significant difference was found in blood pressure, bodyweight and kidney weight/bodyweight between high-salt diet group and normal salt diet group (P>0.05). The expression of OPN mRNA (0.27+/-0.16 vs 0.15+/-0.13, P<0.05) and its protein (0.78+/-0.15 vs 0.61+/-0.11, P<0.01) in SD rat kidney with high-salt diet were up-regulated at the end of 12th week compared with the rats with normal salt diet. CONCLUSIONS: High-salt diet increase the expression of OPN mRNA and its protein in SD rat kidney. Renal injury induced by high salt intake might be independent of blood pressure change. PMID- 15897151 TI - [Anastomosis of the cecum and anus preserving the ileocecal valve through the rectal muscular cuff for benign colorectal diseases]. AB - OBJECTIVE: To evaluate the effect of anastomosis of the cecum and anus preserving the ileocecal valve through the rectal muscular cuff in the treatment of benign colorectal diseases. METHODS: Eight cases including 5 ulcerative colitis and 3 familial adenomatous polyposis treated with this surgical procedure between 1990 to 2000 were retrospectively reviewed. RESULTS: During the 5 to 10 years of follow-up, no relapse, wound infection or anal fistula occurred in these patients, who had normal urination and sexual functions. The bowel movements decreased to 3 to 5 times a day within 8 to 10 weeks after the operation, and recovered to normal status (1 to 3 times a day) in 3-4 months. CONCLUSION: This surgical procedure could produce clinically satisfactory result in the cases of ulcerative colitis and familial adenomatous polyposis. PMID- 15897152 TI - The role of D1 dopamine receptors and phospho-ERK in mediating cytotoxicity. Commentary. AB - Striatal neurodegeneration observed in several neurological diseases, occurs through unknown mechanisms. Recent evidence suggests that its pathogenesis may be linked, in part, to high synaptic levels of dopamine (DA), which can then cause neurotoxicity of striatal neurons through mitogen-activated protein kinases (MAPKs). Here we comment on the role of extracellular signal-regulated kinase (ERK) activation in the cytotoxicity mediated upon activation of the D1 DA receptor, and describe a possible mechanism for phospho-ERK (p-ERK) in inducing cytotoxicity. PMID- 15897153 TI - Role of activated astrocytes in neuronal damage: potential links to HIV-1 associated dementia. AB - HIV-1-associated dementia (HAD) is an important complication of HIV-1 infection. Reactive astrogliosis is a key pathological feature in HAD brains and in other central nervous system (CNS) diseases. Activated astroglia may play a critical role in CNS inflammatory diseases such as HAD. In order to test the hypothesis that activated astrocytes cause neuronal injury, we stimulated primary human fetal astrocytes with HAD-relevant pro-inflammatory cytokine IL-1beta. IL-1beta activated astrocytes induced apoptosis and significant changes in metabolic activity in primary human neurons. An FITC-conjugated pan-caspase inhibitor peptide FITC-VAD-FMK was used for confirming caspase activation in neurons. IL 1beta activation enhanced the expression of death protein FasL in astrocytes, suggesting that FasL is one of the potential factors responsible for neurotoxicity observed in HAD and other CNS diseases involving glial inflammation. Our data presented here add to the developing picture of role of activated glia in HAD pathogenesis. PMID- 15897154 TI - MPTP and SNpc DA neuronal vulnerability: role of dopamine, superoxide and nitric oxide in neurotoxicity. Minireview. AB - Parkinson disease (PD) is a common neurodegenerative disease of unknown origin that is characterized, mainly, by a significant reduction in the number of dopamine neurons in the substantia nigra pars compacta (SNpc) of the brain and a dramatic reduction in dopamine levels in the corpus striatum. For reasons that we do not know, the dopamine neuron seems to be more vulnerable to damage than any other neuron in the brain. Although hypotheses of damage to the dopamine neuron include oxidative stress, growth factor decline, excitotoxicity, inflammation in the SNpc and protein aggregation, oxidative stress in the nigrostriatal dopaminergic system garners a significant amount of attention. In the oxidative stress hypothesis of PD, superoxide, nitric oxide and dopamine all conspire to create an environment that can be detrimental to the dopamine neuron. MPTP (1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine), the tool of choice for investigations into the mechanisms involved in the death of dopamine neurons in PD, has been used extensively in attempts to sort out what happens in and around the dopamine neuron. Herein, we review the roles of dopamine, superoxide and nitric oxide in the demise of the dopamine neuron in the MPTP model of PD as it relates to the death of the dopamine neuron noted in PD. PMID- 15897155 TI - Effects of organophosphorus compounds on ATP production and mitochondrial integrity in cultured cells. AB - Recent studies in vivo and in vitro suggested that mitochondrial dysfunction follows exposure to organophosphorus (OP) esters. As mitochondrial ATP production is important for cellular integrity, ATP production in the presence of OP neurotoxicants was examined in a human neuronal cell line (SH-SY5Y neuroblastoma cells) and primary dorsal root ganglia (DRG) cells isolated from chick embryos and subsequently cultured to achieve maturation with axons. These cell culture systems were chosen to evaluate toxic effects on the mitochondrial respiratory chain associated with exposure to OP compounds that do and do not cause OP induced delayed neuropathy (OPIDN), a disorder preceded by inhibition of neurotoxic esterase (NTE). Concentration- and time-response studies were done in neuroblastoma cells exposed to phenyl saligenin phosphate (PSP) and mipafox, both compounds that readily induce delayed neuropathy in hens, or paraoxon, which does not. Phenylmethylsulfonyl fluoride (PMSF) was included as a non-neuropathic inhibitor of NTE. Purified neuronal cultures from 9 day-old chick embryo DRG were treated for 12 h with 1 microM PSP, mipafox, or paraoxon. In situ evaluation of ATP production measured by bioluminescence assay demonstrated decreased ATP concentrations both in neuroblastoma cells and chick DRG neurons treated with PSP. Mipafox decreased ATP production in DRG but not in SH-SY5Y cells. This low energy state was present at several levels of the mitochondrial respiratory chain, including Complexes I, II, III, and IV, although Complex I was the most severely affected. Paraoxon and PMSF were not effective at all complexes, and, when effective, required higher concentrations than needed for PSP. Results suggest that mitochondria are an important early target for OP compounds, with exposure resulting in depletion of ATP production. The targeting of neuronal, rather than Schwann cell mitochondria in DRG following exposure to PSP and mipafox was verified by loss of the mitochondrial-specific dye, tetramethylrhodamine, in these cells. No such loss was seen in paraoxon exposed neurons isolated from DRG or in Schwann cells treated with any of the test compounds. PMID- 15897156 TI - Progress in Spinobulbar muscular atrophy research: insights into neuronal dysfunction caused by the polyglutamine-expanded androgen receptor. AB - Spinobulbar muscular atrophy (SBMA, Kennedy's disease) results from the dysfunction and degeneration of specific motor and sensory neurons. The underlying cause of this ligand-dependent neurodegenerative disease is expansion of the CAG trinucleotide repeat in the androgen receptor (AR) gene which leads to lengthening of the polyglutamine tract in the AR protein. Recently, the effects of the polyglutamine-expanded AR have been explored in a number of cellular and animal models. Common themes include research on polyglutamine-containing nuclear inclusions and the effect of molecular chaperone overexpression on their formation. In addition, investigations have highlighted the role that abnormal transcriptional regulation, proteasome dysfunction and altered axonal transport may play in disease pathogenesis. These studies suggest a number of potential treatments for restoring neuronal function. One of the most interesting advances in SBMA research has been the creation of mouse models that recapitulate the key features of SBMA progression in men. Lowering testosterone levels in affected transgenic male mice rescued, and even reversed the polyglutamine-induced neuromuscular phenotype, indicating that manipulating androgen levels in men could be of therapeutic benefit. Although the question of why only a distinct subset of neurons is affected by polyglutamine expansion of the AR remains unsolved, future research will provide further insights into the mechanisms contributing to disease progression in SBMA. PMID- 15897157 TI - Association of apolipoprotein J-positive beta-amyloid plaques with dystrophic neurites in Alzheimer's disease brain. AB - Apolipoprotein J (apoJ), also known as clusterin and SP-40,40, binds soluble beta amyloid (Abeta and is up-regulated in the Alzheimer's disease (AD) brain. In the present study we classified apoJ-immunopositive Abeta deposits in AD temporal cortex, and found apoJ-immunoreactive plaques were often associated with dystrophic neurites. Quantitative immunohistochemical analysis of five AD brains showed that 29% of Abeta deposited in the parenchyma was associated with apoJ. Of Abeta deposits with apoJ immunopositivity, 71% were associated with phospho-tau positive dystrophic neurites in the surrounding tissue. Conversely, 64% of phospho-tau-labeled neuritic deposits were labeled with apoJ. ApoJ was found at the core of these deposits, and co-localized with the amyloid staining agent thioflavine-S. To test the direct effects of apoJ on tau metabolism, we treated cells in culture with apoJ-containing conditioned media, and we injected apoJ containing media into the rat hippocampus. Using both systems, we observed increases in levels of tau and phosphorylated tau. Our findings demonstrate that apoJ immunopositivity strongly correlates with the presence of amyloid and associated neuritic dystrophy in the neuropil of AD temporal cortex, and supports a model where extracellular apoJ facilitates the conversion of diffuse Abeta deposits into amyloid and enhances tau phosphorylation in neurites surrounding these of plaques. PMID- 15897158 TI - Retinoic acid isomers protect hippocampal neurons from amyloid-beta induced neurodegeneration. AB - Attenuating amyloid-beta mediated neurodegeneration is of major therapeutic consideration in the potential treatment of Alzheimer disease. Previously, we found that a high dietary consumption of retinoic acid was associated with a reduced incidence of Alzheimer disease. Therefore, in this study, we investigated whether amyloid-beta mediated cell death in primary hippocampal neurons could be prevented by retinoic acid isomers. Our results suggest that retinoic acid isomers, including all-trans retinoic acid, 9-cis retinoic acid, and 13-cis retinoic acid, may play an important role in protecting neurons from amyloid-beta -induced cell death. Retinoic acid may therefore afford a novel therapeutic mechanism for the treatment and prevention of Alzheimer disease. PMID- 15897160 TI - Topical immunomodulator therapy with squaric acid dibutylester (SADBE) is effective treatment for severe alopecia areata (AA): results of an open-label, paired-comparison, clinical trial. AB - Severe alopecia areata (AA) may have a chronic relapsing course and is often resistant to current treatments. OBJECTIVES: The aim of our study was to evaluate whether topical immunotherapy with squaric acid dibutylester (SADBE) is able to improve the course of severe AA and to reduce the severity of relapses. METHODS: Fifty-four patients affected by severe AA treated with SADBE who were followed for a period of at least 2 years were selected as the study group. Data collected were compared with those of a matched control group of 54 patients who did not receive any treatment. Student's t-test, analysis of variance (ANOVA) and Pearson's chi-squared test were utilized for data analysis. RESULTS: At the end of therapy, in comparison with the control group, the treatment group showed a statistically significant (p < 0.001) improvement. At follow-up, there was no significant change in relapse rate (treated 44% vs control 52%). However, relapses in the treated group were significantly less severe compared with the control group (p < 0.001). CONCLUSIONS: Our data suggest that topical SADBE represents a valid therapeutic option in severe AA, and may prove to be disease modifying. PMID- 15897161 TI - Brief case series: montelukast, at doses recommended for asthma treatment, reduces disease severity and increases soluble CD14 in children with atopic dermatitis. AB - BACKGROUND: The choice of oral therapeutic agents for the treatment of atopic dermatitis (AD) in children is limited. Montelukast, a specific cysteinyl leukotriene (LT) receptor antagonist, may be useful in alleviating AD symptoms. OBJECTIVE: To evaluate the clinical and immunological effects of montelukast in children with AD. METHODS: After a 2-week run-in, children with AD were started on oral montelukast 5 mg once-daily for children < 12 years of age and 10 mg for older children. The clinical severity of AD as indicated by the SCORing Atopic Dermatitis (SCORAD) score, and serum soluble CD14 and urinary leukotriene E4 (LTE4) concentrations were evaluated at baseline and the end of a 3-month treatment period. RESULTS: Four boys and three girls, with a median (range) age of 12 (3-16) years, participated in the study. The total SCORAD was reduced in five patients (by 30-84%) and remained similar in two patients. Their median (range) SCORAD scores before and after treatment were 34.7 (16.5-54.8) and 17.0 (6.9-36.9) (p = 0.046). The intensity component of SCORAD also decreased from 5 (2-10) to 3 (1-7) (p = 0.042). Serum sCD14 levels increased significantly from 5533 (4575-6452) ng/ml to 6259 (5617-8988) ng/ml (p = 0.028), whereas urinary LTE4 levels remained the same (p = 0.735). CONCLUSIONS: Montelukast, at doses recommended for asthma treatment, resulted in over 30% reduction in the total SCORAD in some children. Treatment with montelukast may also be associated with deviation of the immune system towards the Th1-specific pathway. PMID- 15897162 TI - Cutaneous photosensitivity induced by paclitaxel and trastuzumab therapy associated with aberrations in the biosynthesis of porphyrins. AB - BACKGROUND: Paclitaxel and trastuzumab are new treatments for patients with metastatic breast cancer. CASE REPORT: We describe here a 40-year-old female patient with metastatic breast cancer who developed a photosensitive rash 1 month after initiation of paclitaxel and trastuzumab therapy. The eruption appeared on the dorsal aspect of her hands, forearms, legs and face and consisted of erythema, edema and vesicles, and was associated with distal onycholysis. Aberrations in various parameters of the metabolism of porphyrins were observed in urine and erythrocytes. Sun avoidance and withdrawal of paclitaxel was followed by resolution of the rash and a return to the normal pattern of porphyrins biosynthesis. CONCLUSION: The combination of paclitaxel and trastuzumab treatment and sun exposure may induce a photosensitive reaction, associated with changes in various parameters of porphyrins biosynthesis. PMID- 15897163 TI - Resting energy expenditure, oxygen consumption and carbon dioxide production during sleep in children with atopic dermatitis. AB - BACKGROUND: Pruritus and scratching are cardinal symptoms of atopic dermatitis (AD). Sleep and growth may also be affected in children with moderate-to-severe AD. We evaluated whether resting energy expenditure (REE), oxygen consumption (VO2) and carbon dioxide production (VCO2) in various stages of sleep were influenced by the disease severity. METHODS: Disease severity was evaluated by the scoring atopic dermatitis (SCORAD) index. All-night polysomnography was performed and REE, VO2 and VCO2 were measured. RESULTS: Twenty children (13 boys and seven girls) with AD and eight controls were recruited. The median overall SCORAD for our AD patients was 36.8. The total sleep efficiency was lower in patients with severe AD than that obtained in the control group (median: 72% versus 88%; p = 0.039). When compared with mild-to-moderate disease (SCORAD40) and controls, REE, VO2 and VCO2 in patients with severe AD (SCORAD > 40) did not differ in sleep stages I and II combined, stages III and IV combined or the rapid eye movement (REM) stage. REE, VO2 and VCO2 in these sleep stages did not show significant correlation with the overall and the three components of the SCORAD scores. CONCLUSIONS: Children with AD do not appear to have significant disturbance in their resting energy consumption, oxygen consumption and carbon dioxide production during sleep. These parameters do not appear to correlate with the symptomatology of pruritus and sleep disturbance. We speculate that deranged metabolism during sleep is unlikely in children with AD. PMID- 15897164 TI - Old fashioned sodium bicarbonate baths for the treatment of psoriasis in the era of futuristic biologics: an old ally to be rescued. AB - BACKGROUND: For centuries, medicated baths have been one of the first lines of treatment for psoriasis. Even today, with sophisticated immunosuppressive treatments available, Dead Sea salts and spa waters are recognized to be beneficial in the management of psoriatic patients. OBJECTIVE: To assess statistically the efficacy of sodium bicarbonate (NaHCO3) baths in psoriasis patients. METHODS: Thirty-one patients with mild-moderate psoriasis were admitted to this study; Nineteen patients were treated with sodium bicarbonate baths and compared with twelve patients who were administered a placebo. Assessments were made on days 0 and 21. RESULTS: Almost all patients who used NaHCO3 reported a statistically valuable improvement. NaHCO3 baths reduced itchiness and irritation; in general, the patients themselves recognized a beneficial impact on their psoriasis, so much so that they have continued to bathe in NaHCO3 even after the end of the study. CONCLUSION: We maintain that even with sophisticated immunosuppressive and highly specific anti-cytokine and anti-chemokines treatments available for the treatment of psoriasis, an older and often forgotten treatment can still play a role. PMID- 15897165 TI - Clobetasol propionate shampoo 0.05% and calcipotriol solution 0.005%: a randomized comparison of efficacy and safety in subjects with scalp psoriasis. AB - BACKGROUND: Scalp involvement in psoriatic patients represents a common issue. Treatment of the hairy skin requires adequate pharmaceutical formulations; hence, a new specific shampoo formulation of clobetasol propionate 0.05% was developed by Galderma R&D, Inc. METHODS: For this multicenter, randomized, investigator masked, parallel group study, 151 subjects with moderate to severe scalp psoriasis were randomized to 4 weeks of treatment with clobetasol propionate shampoo or calcipotriol solution. RESULTS: Clobetasol propionate demonstrated significantly superior efficacy to calcipotriol solution (total severity score: mean difference 0.51, 95% CI 0.05-0.97, p = 0.028; global severity score: mean difference 0.43, 95% CI 0.08-0.78, p = 0.016). Adverse events were more common in the calcipotriol group than in the clobetasol propionate shampoo group. Telangiectasia and skin atrophy did not differ significantly between treatments; however, a burning sensation was significantly more common in the calcipotriol solution group. CONCLUSIONS: Short contact therapy of scalp psoriasis with this new shampoo formulation of clobetasol propionate was significantly more effective and better tolerated than calcipotriol solution for the treatment of scalp psoriasis. PMID- 15897166 TI - Systemic treatment for moderate to severe psoriasis: estimates of failure rates and direct medical costs in a north-eastern US managed care plan. AB - BACKGROUND: Estimates of US medical costs related to psoriasis treatment are limited and tend to understate the economic burden of moderate to severe psoriasis, which often requires the use of systemic agents, phototherapy or both. OBJECTIVE: To estimate treatment failure rates and direct medical costs associated with the use of systemic agents and phototherapy in US patients with psoriasis. METHODS: Claims records from a large New England-based health insurer were used to obtain patient-level data. Eligible patients with at least one claim listing an ICD-9-CM code for psoriasis (696.0; 696.1) were identified. Patients not receiving systemic treatments (methotrexate, cyclosporine, acitretin) or phototherapy (ultraviolet B with or without tar or petrolatum, psoralen and ultraviolet A [PUVA]) were excluded. Treatment failure was defined as a switch in therapy, augmentation with non-topical therapies, discontinuation following uptitration of dose or discontinuation following hospitalization. Medical costs included those related to pharmacy (over-the-counter medication excluded), institutional services (inpatient and outpatient) and professional services. RESULTS: A total of 2068 patients with moderate to severe psoriasis were included in the analysis. Over a 1-year period, approximately 20% of patients experienced treatment failure. The mean time to failure among patients who switched therapy ranged from 3 to 6 months. Mean annual pharmacy costs in the various treatment groups (categorized according to initial therapy received) ranged from 257 dollars to 1992 dollars per patient. Mean annual costs for institutional and professional services ranged from 156 dollars to 799 dollars and 183 dollars to 481 dollars per patient, respectively. The 99th percentile annual pharmacy and institutional costs exceeded 10,000 dollars and 18,000 dollars, respectively. CONCLUSION: Treatment of moderate to severe psoriasis with traditional systemic agents or phototherapy is associated with a high likelihood of treatment failure and a considerable economic burden. PMID- 15897167 TI - Therapeutic options for the treatment of tinea capitis: griseofulvin versus fluconazole. AB - BACKGROUND: Tinea capitis is a relatively common fungal infection of childhood. Griseofulvin has been the mainstay of treatment for many years. However, newer oral antifungal agents are being used more frequently. OBJECTIVE: Our purpose was to evaluate the therapeutic efficacy of fluconazole in comparison with griseofulvin in the treatment of tinea capitis. METHODS: We performed a single blind, randomized, prospective evaluation of 40 patients with a clinical and mycologic diagnosis of tinea capitis. One group received fluconazole for 4 weeks. The other group received griseofulvin for 6 weeks. Five clinical parameters were evaluated. Mycologic examinations were performed at baseline and at the end of 8 weeks. RESULTS: Patients ranged in age from 1 to 16 years; 80% were boys and 20% were girls. Mycologic examinations disclosed Trichophyton verrucosum in 40% of patients, T. violaceum in 40% and Microsporum canis in 20%. At week 8, the griseofulvin-treated group showed a cure rate of 76%, and the fluconazole-treated group 78%. The cure rates were not statistically significant. CONCLUSION: Fluconazole constitutes an alternative but, because of greater availability and lower cost, griseofulvin remains the treatment of choice for tinea capitis. PMID- 15897168 TI - Treatment of chromoblastomycosis with terbinafine: experience with four cases. AB - BACKGROUND: Chromoblastomycosis is a chronic subcutaneous mycosis that occurs more frequently in tropical and subtropical areas and is caused by a group of dematiaceous fungi. It is a difficult-to-treat mycosis with low cure rates and a high rate of relapses. OBJECTIVE: The objective of this trial is to prove the efficacy and tolerance of oral terbinafine in four cases of chromoblastomycosis. METHODS AND RESULTS: We included four cases of chromoblastomycosis, proved clinically and mycologically, that are presented herein; three of them caused by Fonsecaea pedrosoi and one by Phialophora verrucosa. Two had a small extension and the other two were of medium and large extension. Oral terbinafine was administered at a dose of 500 mg/day, which was reduced to half in two of the cases once an 80% improvement had been reached; in the third case the initial dose was maintained, and in the fourth case the dose was increased to 750 mg/day. Three cases reached clinical and mycological cure in a mean treatment period of 7 months, the fourth case reached a significant improvement only after 1.2 years of treatment. The medication was well tolerated; no liver alterations occurred; only one patient suffered mild dyspepsia. CONCLUSION: Terbinafine at 500 mg/day doses represents one of the best treatments for chromoblastomycosis due to its efficacy and excellent tolerability. PMID- 15897169 TI - Combination of fluconazole and urea in a nail lacquer for treating onychomycosis. AB - BACKGROUND: METHODS: Altogether, 13 patients with onychomycosis, aged 25-78 years, most with involvement of the matrix region, were treated. A solution of 1% fluconazole and 20% urea in a mixture of ethanol and water was applied once daily, at bedtime. RESULTS: In four patients there was complete resolution of the condition; four patients who had involvement of one nail only demonstrated a 90% improvement. Of the four patients who had presented with involvement of both big toenails, two showed 50% improvement bilaterally and in the remaining two patients there was a 90% improvement in one nail and a 50% improvement in the other. One patient with proximal white subungual onychomycosis of the finger nail, who grew Alternaria, relapsed during treatment after an initial positive response. CONCLUSION: Generally, a favourable response to local treatment may be anticipated only if the distal two-thirds of the nail is affected. In view of this, and despite the fact that most of the patients were severely affected with involvement of the proximal third of the nail, the response to local therapy was appreciable. PMID- 15897170 TI - Resolution of pruritus secondary to polycythemia vera in a patient treated with narrow-band ultraviolet B phototherapy. AB - Narrow-band ultraviolet B (UVB) is a newer treatment modality for photoresponsive skin diseases. Since its introduction, dermatologists continue to use it for a myriad of dermatoses. Polycythemia vera is one cause of intractable pruritus that has recently been treated successfully with narrow-band UVB. We describe the case of a 77-year-old Caucasian male with a 4-year history of polycythemia vera complicated by intractable pruritus. Narrow-band UVB was successfully used to treat his pruritus. The patient noted an improvement in pruritus within the first four treatments and almost complete resolution after 18 treatments. PMID- 15897171 TI - Treatment of hidradenitis suppurativa with infliximab in a patient with Crohn's disease. AB - BACKGROUND: Hidradenitis suppurativa is a chronic, inflammatory, scarring disease characterized by recurrent flares. Recently, a group of 'autoinflammatory disorders' has been described. These disorders are characterized by recurrent inflammatory episodes not mediated by autoantibodies or antigen-specific T-cells. Some of these autoinflammatory disorders have been successfully treated with anti tumor necrosis factor antibodies. OBSERVATIONS: We describe a patient with hidradenitis suppurativa and a history of inactive Crohn's disease who was treated with infliximab, with subsequent improvement in her skin disease. CONCLUSIONS: This case suggests that infliximab may be effective in the treatment of hidradenitis suppurativa in association with Crohn's disease. We also suggest that hidradenitis suppurativa may be closely linked with this new group of autoinflammatory disorders. PMID- 15897173 TI - Introduction: dissection and simplification of chromosome segregation. PMID- 15897174 TI - How might cohesin hold sister chromatids together? AB - The sister chromatid cohesion essential for the bi-orientation of chromosomes on mitotic spindles depends on a multi-subunit complex called cohesin. This paper reviews the evidence that cohesin is directly responsible for holding sister DNAs together and considers how it might perform this function in the light of recent data on its structure. PMID- 15897175 TI - Diversity and redundancy in bacterial chromosome segregation mechanisms. AB - Bacterial cells are much smaller and have a much simpler overall structure and organization than eukaryotes. Several prominent differences in cell organization are relevant to the mechanisms of chromosome segregation, particularly the lack of an overt chromosome condensation/decondensation cycle and the lack of a microtubule-based spindle. Although bacterial chromosomes have a rather dispersed appearance, they nevertheless have an underlying high level of spatial organization. During the DNA replication cycle, early replicated (oriC) regions are localized towards the cell poles, whereas the late replicated terminus (terC) region is medially located. This spatial organization is thought to be driven by an active segregation mechanism that separates the sister chromosomes continuously as replication proceeds. Comparisons of various well-characterized bacteria suggest that the mechanisms of chromosome segregation are likely to be diverse, and that in many bacteria, multiple overlapping mechanisms may contribute to efficient segregation. One system in which the molecular mechanisms of chromosome segregation are beginning to be elucidated is that of sporulating cells of Bacillus subtilis. The key components of this system have been identified, and their functions are understood, in outline. Although this system appears to be specialized, most of the functions are conserved widely throughout the bacteria. PMID- 15897176 TI - SMC proteins and chromosome mechanics: from bacteria to humans. AB - Chromosome cohesion and condensation are essential prerequisites of proper segregation of genomes during mitosis and meiosis, and are supported by two structurally related protein complexes, cohesin and condensin, respectively. At the core of the two complexes lie members of the structural maintenance of chromosomes (SMC) family of ATPases. SMC proteins are also found in most bacterial and archaeal species, implicating the existence of an evolutionarily conserved theme of higher-order chromosome organization and dynamics. SMC dimers adopt a two-armed structure with an ATP-binding cassette (ABC)-like domain at the distal end of each arm. This article reviews recent work on the bacterial and eukaryotic SMC protein complexes, and discusses current understanding of how these uniquely designed protein machines may work at a mechanistic level. It seems most likely that the action of SMC proteins is highly dynamic and plastic, possibly involving a diverse array of intramolecular and intermolecular protein protein interactions. PMID- 15897177 TI - Shugoshin protects cohesin complexes at centromeres. AB - The different regulation of sister chromatid cohesion at centromeres and along chromosome arms is obvious during meiosis, because centromeric cohesion, but not arm cohesion, persists throughout anaphase of the first division. A protein required to protect centromeric cohesin Rec8 from separase cleavage has been identified and named shugoshin (or Sgo1) after shugoshin ("guardian spirit" in Japanese). It has become apparent that shugoshin shows marginal homology with Drosophila Mei-S332 and several uncharacterized proteins in other eukaryotic organisms. Because Mei-S332 is a protein previously shown to be required for centromeric cohesion in meiosis, it is now established that shugoshin represents a conserved protein family defined as a centromeric protector of Rec8 cohesin complexes in meiosis. The regional difference of sister chromatid cohesion is also observed during mitosis in vertebrates; the cohesion is much more robust at the centromere at metaphase, where it antagonizes the pulling force of spindle microtubules that attach the kinetochores from opposite poles. The human shugoshin homologue (hSgo1) is required to protect the centromeric localization of the mitotic cohesin, Scc1, until metaphase. Bub1 plays a crucial role in the localization of shugoshin to centromeres in both fission yeast and humans. PMID- 15897178 TI - Towards understanding the molecular basis of bacterial DNA segregation. AB - Bacteria ensure the fidelity of genetic inheritance by the coordinated control of chromosome segregation and cell division. Here, we review the molecules and mechanisms that govern the correct subcellular positioning and rapid separation of newly replicated chromosomes and plasmids towards the cell poles and, significantly, the emergence of mitotic-like machineries capable of segregating plasmid DNA. We further describe surprising similarities between proteins involved in DNA partitioning (ParA/ParB) and control of cell division (MinD/MinE), suggesting a mechanism for intracellular positioning common to the two processes. Finally, we discuss the role that the bacterial cytoskeleton plays in DNA partitioning and the missing link between prokaryotes and eukaryotes that is bacterial mechano-chemical motor proteins. PMID- 15897179 TI - Rings, bracelet or snaps: fashionable alternatives for Smc complexes. AB - The mechanism of higher order chromosome organization has eluded researchers for over 100 years. A breakthrough occurred with the discovery of multi-subunit protein complexes that contain a core of two molecules from the structural maintenance of chromosome (Smc) family. Smc complexes are important structural components of chromosome organization in diverse aspects of DNA metabolism, including sister chromatid cohesion, condensation, global gene repression, DNA repair and homologous recombination. In these different processes, Smc complexes may facilitate chromosome organization by tethering together two parts of the same or different chromatin strands. The mechanism of tethering by Smc complexes remains to be elucidated, but a number of intriguing topological alternatives are suggested by the unusual structural features of Smc complexes, including their large coiled-coil domains and ATPase activities. Distinguishing between these possibilities will require innovative new approaches. PMID- 15897180 TI - Merotelic kinetochores in mammalian tissue cells. AB - Merotelic kinetochore attachment is a major source of aneuploidy in mammalian tissue cells in culture. Mammalian kinetochores typically have binding sites for about 20-25 kinetochore microtubules. In prometaphase, kinetochores become merotelic if they attach to microtubules from opposite poles rather than to just one pole as normally occurs. Merotelic attachments support chromosome bi orientation and alignment near the metaphase plate and they are not detected by the mitotic spindle checkpoint. At anaphase onset, sister chromatids separate, but a chromatid with a merotelic kinetochore may not be segregated correctly, and may lag near the spindle equator because of pulling forces toward opposite poles, or move in the direction of the wrong pole. Correction mechanisms are important for preventing segregation errors. There are probably more than 100 times as many PtK1 tissue cells with merotelic kinetochores in early mitosis, and about 16 times as many entering anaphase as the 1% of cells with lagging chromosomes seen in late anaphase. The role of spindle mechanics and potential functions of the Ndc80/Nuf2 protein complex at the kinetochore/microtubule interface is discussed for two correction mechanisms: one that functions before anaphase to reduce the number of kinetochore microtubules to the wrong pole, and one that functions after anaphase onset to move merotelic kinetochores based on the ratio of kinetochore microtubules to the correct versus incorrect pole. PMID- 15897181 TI - Chromosome bi-orientation on the mitotic spindle. AB - For proper chromosome segregation, sister kinetochores must attach to microtubules extending from opposite spindle poles prior to anaphase onset. This state is called sister kinetochore bi-orientation or chromosome bi-orientation. The mechanism ensuring chromosome bi-orientation lies at the heart of chromosome segregation, but is still poorly understood. Recent evidence suggests that mal oriented kinetochore-to-pole connections are corrected in a tension-dependent mechanism. The cohesin complex and the Ipl1/Aurora B protein kinase seem to be key regulators for this correction. In this article, I discuss how cells ensure sister kinetochore bi-orientation for all chromosomes, mainly focusing on our recent findings in budding yeast. PMID- 15897182 TI - Two distinct pathways responsible for the loading of CENP-A to centromeres in the fission yeast cell cycle. AB - CENP-A is a centromere-specific histone H3 variant that is- essential for faithful chromosome segregation in all eukaryotes thus far investigated. We genetically identified two factors, Ams2 and Mis6, each of which is required for the correct centromere localization of SpCENP-A (Cnp1), the fission yeast homologue of CENP-A. Ams2 is a cell-cycle-regulated GATA factor that localizes on the nuclear chromatin, including on centromeres, during the S phase. Ams2 may be responsible for the replication-coupled loading of SpCENP-A by facilitating nucleosomal formation during the S phase. Consistently, overproduction of histone H4, but not that of H3, suppressed the defect of SpCENP-A localization in Ams2 deficient cells. We demonstrated the existence of at least two distinct phases for SpCENP-A loading during the cell cycle: the S phase and the late-G2 phase. Ectopically induced SpCENP-A was efficiently loaded onto the centromeres in G2 arrested cells, indicating that SpCENP-A probably undergoes replication-uncoupled loading after the completion of S phase. This G2 loading pathway of SpCENP-A may require Mis6, a constitutive centromere-binding protein that is also implicated in the Mad2-dependent spindle attachment checkpoint response. Here, we discuss the functional relationship between the flexible loading mechanism of CENP-A and the plasticity of centromere chromatin formation in fission yeast. PMID- 15897183 TI - Basic mechanism of eukaryotic chromosome segregation. AB - We now have firm evidence that the basic mechanism of chromosome segregation is similar among diverse eukaryotes as the same genes are employed. Even in prokaryotes, the very basic feature of chromosome segregation has similarities to that of eukaryotes. Many aspects of chromosome segregation are closely related to a cell cycle control that includes stage-specific protein modification and proteolysis. Destruction of mitotic cyclin and securin leads to mitotic exit and separase activation, respectively. Key players in chromosome segregation are SMC containing cohesin and condensin, DNA topoisomerase II, APC/C ubiquitin ligase, securin-separase complex, aurora passengers, and kinetochore microtubule destabilizers or regulators. In addition, the formation of mitotic kinetochore and spindle apparatus is absolutely essential. The roles of principal players in basic chromosome segregation are discussed: most players have interphase as well as mitotic functions. A view on how the centromere/kinetochore is formed is described. PMID- 15897184 TI - Mechanism and function of poleward flux in Xenopus extract meiotic spindles. AB - In Xenopus extract meiotic spindles, microtubules slide continuously towards their minus ends, a process called poleward flux. This article discusses recent progress in determining the mechanism of poleward flux, and its functions in spindle organization and generating force on chromosomes. Bipolar organization is required for flux and inhibition of the mitotic kinesin Eg5 inhibits flux, suggesting the sliding force for flux is generated by Eg5 pushing anti-parallel microtubules apart. An important function of flux in spindle organization may be to transport minus ends nucleated at chromatin towards the pole. By pulling microtubules through attachment sites at kinetochores, flux may generate poleward force on metaphase chromosomes. PMID- 15897185 TI - Chromosomal instability and human cancer. AB - Genetic instability is a defining feature of human cancer. The main type of genetic instability, chromosomal instability (CIN), enhances the rate of gross chromosomal changes during cell division. CIN is brought about by mutations of CIN genes, i.e. genes that are involved in maintaining the genomic integrity of the cell. A major question in cancer genetics is whether genetic instability is a cause and hence a driving force of tumorigenesis. A mathematical framework for studying the somatic evolution of cancer sheds light onto the causal relations between CIN and human cancer. PMID- 15897186 TI - Explaining the oligomerization properties of the spindle assembly checkpoint protein Mad2. AB - Mad2 is an essential component of the spindle assembly checkpoint (SAC), a molecular device designed to coordinate anaphase onset with the completion of chromosome attachment to the spindle. Capture of chromosome by microtubules occur on protein scaffolds known as kinetochores. The SAC proteins are recruited to kinetochores in prometaphase where they generate a signal that halts anaphase until all sister chromatid pairs are bipolarly oriented. Mad2 is a subunit of the mitotic checkpoint complex, which is regarded as the effector of the spindle checkpoint. Its function is the sequestration of Cdc20, a protein required for progression into anaphase. The function of Mad2 in the checkpoint correlates with a dramatic conformational rearrangement of the Mad2 protein. Mad2 adopts a closed conformation (C-Mad2) when bound to Cdc20, and an open conformation (O-Mad2) when unbound to this ligand. Checkpoint activation promotes the conversion of O-Mad2 to Cdc20-bound C-Mad2. We show that this conversion requires a C-Mad2 template and we identify this in Mad1-bound Mad2. In our proposition, Mad1-bound C-Mad2 recruits O-Mad2 to kinetochores, stimulating Cdc20 capture, implying that O-Mad2 and C-Mad2 form dimers. We discuss Mad2 oligomerization and link our discoveries to previous observations related to Mad2 oligomerization. PMID- 15897187 TI - ARF6 GTPase controls bacterial invasion by actin remodelling. AB - The obligate intracellular bacterium Chlamydia penetrates the host epithelial cell by inducing cytoskeleton and membrane rearrangements reminiscent of phagocytosis. Here we report that Chlamydia induces a sharp and transient activation of the endogenous small GTP-binding protein ARF6, which is required for efficient uptake. We also show that a downstream effector of ARF6, phosphatidylinositol 4-phosphate 5-kinase and its product, phosphatidylinositol 4,5-bisphosphate were instrumental for bacterial entry. By contrast, ARF6 activation of phospholipase D was not required for Chlamydia uptake. ARF6 activation was necessary for extensive actin reorganization at the invasion sites. Remarkably, these signalling players gathered with F-actin in a highly organized three-dimensional concentric calyx-like protrusion around invasive bacteria. These results indicate that ARF6, which controls membrane delivery during phagocytosis of red blood cells in macrophages, has a different role in the entry of this small bacterium, controlling cytoskeletal reorganization. PMID- 15897188 TI - Structure-guided saturation mutagenesis of N-acetylneuraminic acid lyase for the synthesis of sialic acid mimetics. AB - Analogues of N-acetylneuraminic acid (sialic acid, NANA, Neu5Ac), including 6 dipropylcarboxamides, have been found to be selective and potent inhibitors of influenza sialidases. Sialic acid analogues are, however, difficult to synthesize by traditional chemical methods and the enzyme N-acetylneuraminic acid lyase (NAL) has previously been used for the synthesis of a number of analogues. The activity of this enzyme towards 6-dipropylcarboxamides is, however, low. Here, we used structure-guided saturation mutagenesis to produce variants of NAL with improved activity and specificity towards 6-dipropylcarboxamides. Three residues were targeted for mutagenesis, Asp191, Glu192 and Ser208. Only substitution at position 192 produced significant improvements in activity towards the dipropylamide. One variant, E192N, showed a 49-fold improvement in catalytic efficiency towards the target analogue and a 690-fold shift in specificity from sialic acid towards the analogue. These engineering efforts provide a scaffold for the further tailoring of NAL for the synthesis of sialic acid mimetics. PMID- 15897189 TI - Biological, biochemical, and kinetic effects of mutations of the cardiomyopathy loop of Dictyostelium myosin II: importance of ALA400. AB - The cardiomyopathy (CM)-loop of the heavy chain of class-II myosins begins with a highly conserved Arg residue (whose mutation in human beta-cardiac myosin II results in familial hypertrophic cardiomyopathy). The CM-loop of Dictyostelium myosin II (Arg397-Gln407) is essential for its biological functions and biochemical activities. We found that the CM-loop of smooth muscle myosin II substituted partially, and the CM-loop of beta-cardiac myosin II less well, for growth, capping of surface receptors and development, and the actin-activated MgATPase and in vitro motility activities of purified myosins. There was little correlation between the biochemical and biological activities of the two chimeras and 19 point mutants, but only the five mutants with k cat/K actin values equivalent to wild-type myosin supported essentially full biological function. The three point mutations of Arg397 equivalent to those that result in hypertrophic cardiomyopathy in humans had minimal biological effects and different biochemical effects. The A400V mutation rendered full-length wild-type myosin almost completely inactive, both in vitro and in vivo, and the reverse V400A mutation in the cardiac CM-loop chimera restored almost full activity, even though the sequence still differed from wild-type in 7 of 11 positions. Transient kinetic studies of acto-subfragment-1 (S1) showed that the chimeras and the Ala/Val, Val/Ala mutations do not affect the equilibrium or the association and dissociation rate constants for either ATP or ADP binding to acto-S1 or the rate of ATP-induced dissociation of acto-S1. We conclude that the Ala/Val, Val/Ala mutations affect the release of Pi from acto-S1.ADP.Pi. In addition, Val at position 400 substantially reduces the affinity of actin for S1 in the absence of nucleotide. PMID- 15897191 TI - Regulatory mechanism of histidine-tagged homocitrate synthase from Saccharomyces cerevisiae. II. Theory. AB - In this study, rate equations that predict the regulatory kinetic behavior of homocitrate synthase were derived, and simulation of the predicted behavior was carried out over a range of values for the kinetic parameters. The data obtained allow application of the resulting expressions to enzyme systems that exhibit activation and inhibition as a result of the interaction of effectors at multiple sites in the free enzyme. Homocitrate synthase was used as an example in terms of its activation by Na+ binding to the active enzyme conformer at an allosteric site, inhibition by binding to the active site, and inhibition by lysine binding to the less active enzyme conformer. PMID- 15897190 TI - LIM kinase 1 coordinates microtubule stability and actin polymerization in human endothelial cells. AB - Microtubule (MT) destabilization promotes the formation of actin stress fibers and enhances the contractility of cells; however, the mechanism involved in the coordinated regulation of MTs and the actin cytoskeleton is poorly understood. LIM kinase 1 (LIMK1) regulates actin polymerization by phosphorylating the actin depolymerization factor, cofilin. Here we report that LIMK1 is also involved in the MT destabilization. In endothelial cells endogenous LIMK1 co-localizes with MTs and forms a complex with tubulin via the PDZ domain. MT destabilization induced by thrombin or nocodazole resulted in a decrease of LIMK1 colocalization with MTs. Overexpression of wild type LIMK1 resulted in MT destabilization, whereas the kinase-dead mutant of LIMK1 (KD) did not affect MT stability. Importantly, down-regulation of endogenous LIMK1 by small interference RNA resulted in abrogation of the thrombin-induced MTs destabilization and the inhibition of thrombin-induced actin polymerization. Expression of Rho kinase 2, which phosphorylates and activates LIMK1, dramatically decreases the interaction of LIMK1 with tubulin but increases its interaction with actin. Interestingly, expression of KD-LIMK1 or small interference RNA-LIMK1 prevents thrombin-induced microtubule destabilization and F-actin formation, suggesting that LIMK1 activity is required for thrombin-induced modulation of microtubule destabilization and actin polymerization. Our findings indicate that LIMK1 may coordinate microtubules and actin cytoskeleton. PMID- 15897192 TI - Regulatory mechanism of histidine-tagged homocitrate synthase from Saccharomyces cerevisiae. I. Kinetic studies. AB - Homocitrate synthase (HCS) catalyzes one of the regulated steps of the alpha aminoadipate pathway for lysine biosynthesis in fungi. The kinetic mechanism of regulation of HCS from Saccharomyces cerevisiae by Na+ and the feedback inhibitor lysine was studied by measuring the initial rate in the absence and presence of the effectors. The data suggest that Na+ is an activator at low concentrations and an inhibitor at high concentrations and that these effects occur as a result of the monovalent ion binding to two different sites in the free enzyme. Inhibition and activation by Na+ can occur simultaneously, with the net rate of the enzyme determined by Na+/K(iNa+) and Na+/K(act), where K(iNa+) and K(act) are the inhibition and activation constants, respectively. The inhibition by Na+ was eliminated at high concentrations of acetyl-CoA, the second substrate bound, but the activation remained. Fluorescence binding studies indicated that lysine bound with high affinity to its binding site as an inhibitor. The inhibition by lysine was competitive versus alpha-ketoglutarate and linear in the physiological range of lysine concentrations up to 5 mm. The effects of Na+ and lysine were independent of one another. A model is developed for regulation of HCS that takes into account all of the effects discussed above. PMID- 15897193 TI - Lipid droplets gain PAT family proteins by interaction with specialized plasma membrane domains. AB - Proteins of the PAT family, named after perilipin, adipophilin, and TIP47 (tail interacting protein of 47 kDa), are associated with lipid droplets and have previously been localized by immunofluorescence microscopy exclusively to the droplet surface. These proteins are considered not to be present in any other subcellular compartment. By applying the high resolution technique of freeze fracture electron microscopy combined with immunogold labeling, we now demonstrate that in macrophages and adipocytes PAT family proteins are, first, distributed not only in the surface but also throughout the lipid droplet core and, second, are integral components of the plasma membrane. Under normal culture conditions these proteins are dispersed in the cytoplasmic leaflet of the plasma membrane. Stimulation of lipid droplet formation by incubation of the cells with acetylated low density lipoprotein leads to clustering of the PAT family proteins in raised plasma membrane domains. Fractures penetrating beneath the plasma membrane demonstrate that lipid droplets are closely apposed to these domains. A similar distribution pattern of labeling in the form of linear aggregates within the clusters is apparent in the cytoplasmic monolayer of the plasma membrane and the immediately adjacent outer monolayer of the lipid droplet. The aggregation of the PAT family proteins into such assemblies may facilitate carrier-mediated lipid influx from the extracellular environment into the lipid droplet. Lipid droplets appear to acquire their PAT proteins by interaction with plasma membrane domains enriched in these proteins. PMID- 15897194 TI - Substrate specificity and recognition is conferred by the pleckstrin homology domain of the Dbl family guanine nucleotide exchange factor P-Rex2. AB - Dbl family guanine nucleotide exchange factors (GEFs) are characterized by the presence of a catalytic Dbl homology domain followed invariably by a lipid binding pleckstrin homology (PH) domain. To date, substrate recognition and specificity of this family of GEFs has been reported to be mediated exclusively via the Dbl homology domain. Here we report the novel and unexpected finding that, in the Dbl family Rac-specific GEF P-Rex2, it is the PH domain that confers substrate specificity and recognition. Moreover, the beta3beta4 loop of the PH domain of P-Rex2 is the determinant for Rac1 recognition, as substitution of the beta3beta4 loop of the PH domain of Dbs (a RhoA- and Cdc42-specific GEF) with that of P-Rex2 confers Rac1-specific binding capability to the PH domain of Dbs. The contact interface between the PH domain of P-Rex2 and Rac1 involves the switch loop and helix 3 of Rac1. Moreover, substitution of helix 3 of Cdc42 with that of Rac1 now enables the PH domain of P-Rex2 to bind this Cdc42 chimera. Despite having the ability to recognize this chimeric Cdc42, P-Rex2 is unable to catalyze nucleotide exchange on Cdc42, suggesting that recognition of substrate and catalysis are two distinct events. Thus substrate recognition can now be added to the growing list of functions that are being attributed to the PH domain of Dbl family GEFs. PMID- 15897195 TI - Viscosity effects on eukaryotic nitrate reductase activity. AB - Rate-limiting processes of catalysis by eukaryotic molybdenum-containing nitrate reductase (NaR, EC 1.7.1.1-3) were investigated using two viscosogens (glycerol and sucrose) and observing their impact on NAD(P)H:NaR activity of corn leaf NaR and recombinant Arabidopsis and yeast NaR. Holo-NaR has two "hinge" sequences between stably folded regions housing its internal electron carriers: 1) Hinge 1 between the molybdenum-containing nitrate reducing module and cytochrome b domain containing heme and 2) Hinge 2 between cytochrome b and cytochrome b reductase (CbR) module containing FAD. Solution viscosity negatively impacted the activity of these holo-NaR forms, which suggests that the rate-limiting events in catalysis were likely to involve large conformational changes that restrict or "gate" internal electron-proton transfers (IET). Little effect of viscosity was observed on recombinant CbR module and methyl viologen nitrate reduction by holo NaR, suggesting that these activities involved no large conformational changes. To determine whether Hinge 2 is involved in gating the first step in IET, the effects of viscosogen on cytochrome c and ferricyanide reductase activities of holo-NaR and ferricyanide reductase activity of the recombinant molybdenum reductase module (CbR, Hinge 2, and cytochrome b) were analyzed. Solution viscosity negatively impacted these partial activities, as if Hinge 2 were involved in gating IET in both enzyme forms. We concluded that both Hinges 1 and 2 appear to be involved in gating IET steps by restricting the movement of the cytochrome b domain relative to the larger nitrate-reducing and electron-donating modules of NaR. PMID- 15897196 TI - Incorporation of factor Va into prothrombinase is required for coordinated cleavage of prothrombin by factor Xa. AB - Prothrombin is activated to thrombin by two sequential factor Xa-catalyzed cleavages, at Arg271 followed by cleavage at Arg320. Factor Va, along with phospholipid and Ca2+, enhances the rate of the process by 300,000-fold, reverses the order of cleavages, and directs the process through the meizothrombin pathway, characterized by initial cleavage at Arg320. Previous work indicated reduced rates of prothrombin activation with recombinant mutant factor Va defective in factor Xa binding (E323F/Y324F and E330M/V331I, designated factor VaFF/MI). The present studies were undertaken to determine whether loss of activity can be attributed to selective loss of efficiency at one or both of the two prothrombin-activating cleavage sites. Kinetic constants for the overall activation of prothrombin by prothrombinase assembled with saturating concentrations of recombinant mutant factor Va were calculated, prothrombin activation was assessed by SDS-PAGE, and rate constants for both cleavages were analyzed from the time course of the concentration of meizothrombin. Prothrombinase assembled with factor VaFF/MI had decreased k(cat) for prothrombin activation with Km remaining unaffected. Prothrombinase assembled with saturating concentrations of factor VaFF/MI showed significantly lower rate for cleavage of plasma-derived prothrombin at Arg320 than prothrombinase assembled with saturating concentrations of wild type factor Va. These results were corroborated by analysis of cleavage of recombinant prothrombin mutants rMz-II (R155A/R284A/R271A) and rP2-II (R155A/R284A/R320A), which can be cleaved only at Arg320 or Arg271, respectively. Time courses of these mutants indicated that mutations in the factor Xa binding site of factor Va reduce rates for both bonds. These data indicate that the interaction of factor Xa with the heavy chain of factor Va strongly influences the catalytic activity of the enzyme resulting in increased rates for both prothrombin-activating cleavages. PMID- 15897197 TI - Novel 3'-ribonuclease and 3'-phosphatase activities of the bacterial non homologous end-joining protein, DNA ligase D. AB - Pseudomonas aeruginosa DNA ligase D (PaeLigD) exemplifies a family of bacterial DNA end-joining proteins that consist of a ligase domain fused to a polymerase domain and a putative nuclease module. The LigD polymerase preferentially adds single ribonucleotides at blunt DNA ends and, as we show here, is also capable of adding up to 4 ribonucleotides to a DNA primer-template. We report that PaeLigD has an intrinsic ability to resect the short tract of 3'-ribonucleotides of a primer-template substrate to the point at which the primer strand has a single 3' ribonucleotide remaining. The failure to digest beyond this point reflects a requirement for a 2'-OH group on the penultimate nucleoside of the primer strand. Replacing the 2'-OH by a 2'-F, 2'-NH2, 2'-OCH3, or 2'-H abolishes the resection reaction. The ribonucleotide resection activity resides within a 187-amino acid N terminal nuclease domain and is the result of at least two component steps: (i) the 3'-terminal nucleoside is first removed to yield a primer strand with a ribonucleoside 3'-PO4 terminus, and (ii) the 3'-PO4 is hydrolyzed to a 3'-OH. The 3'-ribonuclease and 3'-phosphatase activities are both dependent on a divalent cation, specifically manganese. PaeLigD preferentially remodels the 3'-ends of a duplex primer-template substrate rather than a single strand of identical composition, and it prefers DNA primer strands containing a short 3' ribonucleotide tract to an all-RNA primer. The nuclease domain of PaeLigD and its bacterial homologs has no apparent structural or mechanistic similarity to previously characterized nucleases. Thus, we surmise that it exemplifies a novel phosphoesterase family, defined in part by conserved residues Asp-50, Arg-52, and His-84, which are essential for the 3'-ribonuclease and 3'-phosphatase reactions. PMID- 15897198 TI - A superhelical spiral in the Escherichia coli DNA gyrase A C-terminal domain imparts unidirectional supercoiling bias. AB - DNA gyrase is unique among type II topoisomerases in that its DNA supercoiling activity is unidirectional. The C-terminal domain of the gyrase A subunit (GyrA CTD) is required for this supercoiling bias. We report here the x-ray structure of the Escherichia coli GyrA-CTD (Protein Data Bank code 1ZI0). The E. coli GyrA CTD adopts a circular-shaped beta-pinwheel fold first seen in the Borrelia burgdorferi GyrA-CTD. However, whereas the B. burgdorferi GyrA-CTD is flat, the E. coli GyrA-CTD is spiral. DNA relaxation assays reveal that the E. coli GyrA CTD wraps DNA inducing substantial (+) superhelicity, while the B. burgdorferi GyrA-CTD introduces a more modest (+) superhelicity. The observation of a superhelical spiral in the present structure and that of the Bacillus stearothermophilus ParC-CTD structure suggests unexpected similarities in substrate selectivity between gyrase and Topo IV enzymes. We propose a model wherein the right-handed ((+) solenoidal) wrapping of DNA around the E. coli GyrA CTD enforces unidirectional (-) DNA supercoiling. PMID- 15897199 TI - The contribution of RING and B-box 2 domains to retroviral restriction mediated by monkey TRIM5alpha. AB - TRIM5alpha is a cytoplasmic protein that mediates a post-entry block to infection by some retroviruses. TRIM5alpha contains a tripartite motif (TRIM), which includes RING, B-box 2, and coiled-coil domains, and a C-terminal B30.2 (SPRY) domain. We investigated the contribution of the RING and B-box 2 domains to the antiretroviral activity of rhesus monkey TRIM5alpha (TRIM5alpharh), which potently restricts infection by human immunodeficiency virus, type 1 (HIV-1) and simian immunodeficiency virus of African green monkeys (SIVagm). Disruption of the RING domain caused mislocalization of TRIM5alpharh so that the cytoplasmic level of the protein was decreased compared with that of the wild-type protein. Nonetheless, partial ability to restrict HIV-1 and SIVagm was retained by the RING domain mutants. By contrast, although TRIM5alpharh mutants with disrupted B box 2 domains were efficiently expressed and correctly localized to the cytoplasm, antiretroviral activity was absent. The B-box 2 mutants colocalized and associated with wild-type TRIM5alpharh and exerted dominant-negative effects on the antiretroviral activity of the wild-type protein. Taken together with other data, these results indicate that functionally defective TRIM5alpharh molecules that retain a coiled coil can act as dominant-negative inhibitors of wild-type TRIM5alpharh function. The RING domain of TRIM5alpharh is not absolutely required for retrovirus restriction but can influence cytoplasmic levels of the protein and thus indirectly alter function. The B-box 2 domain, by contrast, appears to be essential for efficient retrovirus restriction. PMID- 15897200 TI - The oligomeric T4 primase is the functional form during replication. AB - Replisome DNA primases are responsible for the synthesis of short RNA primers required for the initiation of repetitive Okazaki fragment synthesis on the lagging strand during DNA replication. In bacteriophage T4, the primase (gp61) interacts with the helicase (gp41) to form the primosome complex, an interaction that greatly stimulates the priming activity of gp61. Because gp41 is hexameric, a question arises as to whether gp61 also forms a hexameric structure during replication. Several results from this study support such a structure. Titration of the primase/single-stranded DNA binding followed by fluorescence anisotropy implicated a 6:1 stoichiometry. The observed rate constant, k(cat), for priming was found to increase with the primase concentration, implicating an oligomeric form of the primase as the major functional species. The generation of hetero oligomeric populations of the hexameric primase by controlled mixing of wild type and an inactive mutant primase confirmed the oligomeric nature of the most active primase form. Mutant primases defective in either the N- or C-terminal domains and catalytically inactive could be mixed to create oligomeric primases with restored catalytic activity suggesting an active site shared between subunits. Collectively, these results provide strong evidence for the functional oligomerization of gp61. The potential roles of gp61 oligomerization during lagging strand synthesis are discussed. PMID- 15897201 TI - Structural insights into the mechanism of nuclease A, a betabeta alpha metal nuclease from Anabaena. AB - Nuclease A (NucA) is a nonspecific endonuclease from Anabaena sp. capable of degrading single- and double-stranded DNA and RNA in the presence of divalent metal ions. We have determined the structure of the delta(2-24),D121A mutant of NucA in the presence of Zn2+ and Mn2+ (PDB code 1ZM8). The mutations were introduced to remove the N-terminal signal peptide and to reduce the activity of the nonspecific nuclease, thereby reducing its toxicity to the Escherichia coli expression system. NucA contains a betabeta alpha metal finger motif and a hydrated Mn2+ ion at the active site. Unexpectedly, NucA was found to contain additional metal binding sites approximately 26 A apart from the catalytic metal binding site. A structural comparison between NucA and the closest analog for which structural data exist, the Serratia nuclease, indicates several interesting differences. First, NucA is a monomer rather than a dimer. Second, there is an unexpected structural homology between the N-terminal segments despite a poorly conserved sequence, which in Serratia includes a cysteine bridge thought to play a regulatory role. In addition, although a sequence alignment had suggested that NucA lacks a proposed catalytic residue corresponding to Arg57 in Serratia, the structure determined here indicates that Arg93 in NucA is positioned to fulfill this role. Based on comparison with DNA-bound nuclease structures of the betabeta alpha metal finger nuclease family and available mutational data on NucA, we propose that His124 acts as a catalytic base, and Arg93 participates in the catalysis possibly through stabilization of the transition state. PMID- 15897204 TI - Limitations of Wright formula estimates of renal function. PMID- 15897205 TI - Water relations of the pine exine. AB - BACKGROUND AND AIMS: Water adhesion forces, water absorption capacity and permeability of the pine exine were investigated to consider a possible function of sporopollenin coatings in the control of water transport. METHODS: The experiments were carried out with sporopollenin capsules obtained from pine pollen consisting of an empty central capsule and two sacci. Changes in the concentration of excluded dextran molecules in the medium were analysed to quantify water absorption by purified exine fragments and the osmotic volume flow out of the intact central capsule. KEY RESULTS: The contact angle of sporopollenin to water is higher than the one to ethanol and lower than the one to n-heptane. The water-filled pore space in pine sporopollenin amounts to only 20.6 % of the matrix volume. A monosaccharide was excluded from 15 % and a trisaccharide from about 38 % of this space. Shrinkage of the central capsule induced by permeable osmotica was transient, whereas that induced by sodium polyacrylate (2100 g mol(-1)) was stable. Values obtained for the hydraulic conductance L(P) of the exine (0.39-0.48 microm s(-1) MPa(-1)) are comparable in size to those of biomembranes. Sodium sulfate solutions induced a significant osmotic flow through the exine (reflection coefficient at least 0.6). The exine around the central capsule can be ruptured by equilibration of its lumen with a concentrated electrolyte solution and subsequent transfer to water. The denatured protoplast along with the intact intine was ejected when pollen grains were subjected to this osmotic shock treatment. CONCLUSIONS: The pine exine is easily wetted with water and does not represent a significant barrier to water exchange either liquid or gaseous. Through osmotic burst, it can be separated from the intine. The effect of salts and small solute molecules on water fluxes may be functionally significant for rehydration upon pollination. PMID- 15897206 TI - Soybean genotypic difference in growth, nutrient accumulation and ultrastructure in response to manganese and iron supply in solution culture. AB - BACKGROUND AND AIMS: The objective of this research was to characterize the physiology and cell ultrastructure of two soybean genotypes subjected to nutrient solutions with increasing concentrations of manganese (Mn) at two contrasting iron (Fe) concentrations. Genotypes 'PI227557' and 'Biloxi' were selected based on their distinctly different capacities to accumulate Mn and Fe. * METHODS: Bradyrhizobium-inoculated plants were grown in hydroponic cultures in a greenhouse. Nutrient solutions were supplied with Mn concentrations ranging from 0.3 to 90 microm, at either 5 or 150 microm Fe as FeEDTA. * KEY RESULTS: For both genotypes and at both Fe concentrations, Mn concentrations from 6.6 to 50 microm did not affect shoot, root and nodule mass, or leaf and nodule ureide concentration. Mn concentrations of 70 and 90 microm did not result in visible toxicity symptoms, but hindered growth and nodulation of 'Biloxi'. An Mn concentration of 0.3 microm was, however, deleterious to growth and nodulation for both genotypes, and caused an accumulation of ureides in leaves and major alterations in the ultrastructure of chloroplasts, nuclei and mitochondria, regardless of the Fe concentration. In 'PI227557', there was also a proliferation of Golgi apparatus and endoplasmic reticulum in the cytoplasm of leaf cells, and nodules showed disrupted symbiosomes lacking poly-beta-hydroxybutirate grains concomitantly with a proliferation of endoplasmic reticulum as well as arrested bacterial division. At 15 microm Fe, ferritin-like crystals were formed in the lumen of chloroplasts of 'PI227557' plants. For both genotypes, there was an antagonism between the Fe and Mn concentrations in leaves, the higher values of both microelements being detected in 'PI227557'. The absence of any detectable relationship between Fe or Mn and zinc, phosphorus and copper concentrations in leaves ruled out those micronutrients as relevant for Mn and Fe nutrition in soybeans. * CONCLUSIONS: The results confirmed the greater capacity of 'PI227557' for Mn and Fe accumulation than 'Biloxi' for most nutrient treatments. Hence, 'PI227557' may be a very useful genetic resource both in developing soybean cultivars for growth on low nutrient soils and in physiological studies to understand differing approaches to nutrient accumulation in plants. PMID- 15897207 TI - The worried young adult as a primary care patient. AB - BACKGROUND: Being worried about one's complaint is common among primary health care patients. Persistent and intensive worry may, however, have negative consequences. OBJECTIVES: We explored complaint-related worry and factors associated with it among 18- to 39-year-old primary health care patients. METHODS: Sixty-two patients evaluated the intensity of their worry and the severity of their complaint before seeing their GP. They were also interviewed about their background and filled in questionnaires about general tendency to illness-related worry and psychiatric symptoms. RESULTS: The intensity of worry varied greatly. One fourth of the patients reported intense worry. A general tendency to worry about health and hostile reactions were associated with both the intensity of worry and the severity appraisals. The patient's education and the duration and perceived course of the complaint also played a role in worrying and in the perceptions of the severity of the complaint. CONCLUSIONS: Some psychological characteristics may dispose patients to intensive worrying and pessimistic appraisals of their complaint. This challenges the GP to pay attention to the patients' perspectives and knowledge. Careful elucidation of the patients' experiences of their complaints is especially indicated in the case of complaints of long duration and a stable course. PMID- 15897208 TI - Explanations, explanations, explanations: how do patients with limited English construct narrative accounts in multi-lingual, multi-ethnic settings, and how can GPs interpret them? AB - BACKGROUND: The gap is widening between understanding the subtle ways patients and GPs manage their talk, and superficial discussion of the 'language barrier' among linguistic minority patients. All patients have to explain themselves, not just those for whom English is their first or main language. Patients' explanations reflect how they want the doctor to perceive them as a patient and as a person: they reveal patients' identities. Yet interpretations are not easy when patients' style of talking English is influenced by their first language and cultural background. OBJECTIVE: To explore in detail how patients with limited English and GPs jointly overcome misunderstandings in explanations. METHODS: Using discourse analysis and conversation analysis, we examine how GPs and their patients with limited English negotiate explanations and collaborate to manage, repair or prevent understanding problems. RESULTS: 31% of patients said English was not their first language. Misunderstandings arise owing to a range of linguistic and cultural factors, including stress and intonation patterns, vocabulary, the way a patient sequences their narrative, and patient and GP pursuing different agendas. CONCLUSION: When talk itself is the problem, patients' explanations can lead to misunderstandings, which GPs have to repair if they cannot prevent. Careful interpretation by skillful GPs can reveal patients' knowledge, experience and perspective. PMID- 15897209 TI - Primary care physicians in Hong Kong and Canada--how did their practices differ during the SARS epidemic? AB - BACKGROUND: Hong Kong and Canada have very different primary health care systems. We thus hypothesized family physicians between the two places would be different in how they protected themselves, their staff and families during the epidemic. OBJECTIVE: The purpose of this study was to explore and contrast the impact of SARS on family physicians in Hong Kong and Toronto. METHODS: A postal questionnaire was designed and sent to 183 tutors affiliated with the Chinese University of Hong Kong with 137 replies (74.8%). In Toronto, 150 questionnaires were sent to academic family physicians affiliated with the University of Toronto with 51 replies (34%). RESULTS: All agreed SARS had changed their clinical behaviour. For public health measures in the control of SARS, Hong Kong physicians were less likely to quarantined themselves (77.1% versus 19.4%, P < 0.01) or gave quarantine leave to staff (95% versus 59.7%, P < 0.01) after exposure to probable or suspected SARS. However, they were more likely to wear a mask (52.7% versus 97.7%, P = 0) during consultation, having support staff to wear masks (68.6% versus 97.8%, P = 0) and test patient's temperature (47.1% versus 68.1%, P < 0.01). CONCLUSION: There were noticeable differences in how family physicians deal with SARS between the two cities. As SARS emerged as a global disease, better understanding of practice differences among physicians from different countries would facilitate globalization of public health. PMID- 15897210 TI - The prevalence of symptoms and consultations in pre-school children in the Avon Longitudinal Study of Parents and Children (ALSPAC): a prospective cohort study. AB - BACKGROUND: Pre-school children are frequent users of health services, but little contemporary data are available describing their symptoms or why they consult. OBJECTIVE: To describe symptom and consultation prevalence in pre-school children and to identify the socio-demographic factor or factors associated with consultations for those symptoms. METHODS: Prospective cohort study of 13,617 pre school children living in south-west England. Parents completed questionnaires asking about symptoms and consultations for those symptoms at six, 18, 30, 42 and 57 months. RESULTS: During the pre-school years, all children experienced one or more symptoms, most commonly cold, cough, high temperature, vomiting or diarrhoea. Ninety seven percent consulted a doctor at least once, most commonly for cough, high fever and/or earache. Lower parity was most strongly and consistently associated with higher consultation rates. CONCLUSIONS: Fever, respiratory and gastro-intestinal symptoms are a normal part of pre-school life. Research of acute conditions in young children could focus on the most common symptoms leading to consultation, namely cough, fever and earache. Efforts to support parents' help seeking decision making might usefully be targeted at first time parents. PMID- 15897211 TI - A randomised controlled trial of brief training in assessment and treatment of somatisation: effects on GPs' attitudes. AB - BACKGROUND: Somatising patients frequently present in primary care but GPs often express frustration in dealing with them. A negative attitude may result in missed diagnoses and ineffective treatment. OBJECTIVE: This study aimed to evaluate the effect of a novel, multifaceted training programme on GPs' attitudes towards somatisation. METHODS: The study was performed as a cluster randomised controlled trial with practices as randomisation unit and with a follow-up period of 12 months. Forty-three GPs from 27 practices in Vejle County, Denmark participated. The intervention consisted of a cognitive-oriented educational programme on assessment, treatment and management of somatisation (The Extended Reattribution and Management Model). Outcome measures were GPs' attitudes toward somatoform disorder and somatisation in general measured by the means of questionnaires at baseline and follow-up. The primary outcome was a change in response. RESULTS: Baseline values confirmed previous findings that GPs find it difficult to deal with somatising patients. Compared with the control doctors, intervention doctors' attitudes towards patients with somatoform disorders had changed significantly 12 months after training on the parameters enjoyment (P = 0.008) and anxiety (P = 0.002). Doctors also felt more comfortable in dealing with somatising patients in general (P = 0.002). Attitudes about other parameters related to the doctors feelings, aetiology and course of somatisation changed in the expected direction, but these changes were not statistically significant. CONCLUSION: A brief multifaceted training programme focussing on somatisation was accompanied by a significant change in GPs' attitude towards patients with somatoform disorders. PMID- 15897212 TI - Screening for common mental disorders: who will benefit? Results from a randomised clinical trial. AB - BACKGROUND: Outcomes of studies on mental health screening in primary care are conflicting. A feasible and effective case-finding approach could benefit both GPs and their patients. OBJECTIVES: (1) to examine the effect of using a composite screening questionnaire (SQ) on GPs' recognition and provision of care, and (2) to outline useful strategies for case-finding. METHODS: 38 GPs in Aarhus County, Denmark, volunteered to participate in this trial. 1785 consecutive patients aged 18-65 years consulting with new health problems were included. Patients were screened before consultation using an SQ including scales for somatisation, anxiety, depression and alcohol abuse. Patients were randomised into one of two groups: 900 questionnaires were disclosed to and scored by GPs, 885 were blinded. Number of diagnoses, subjects of conversation, and actions taken were analysed. Additional analyses aimed to identify GP and patient factors that could predict improved outcomes. RESULTS: Overall, disclosure of SQ results increased GPs' recognition of mental disorders by 3.8% [95% confidence interval (CI) -0.5% to 8.0%], and 6.6% (95% CI 1.2% to 12.0%) for patients screened positive. There was a marked variation in GPs' detection rates, and for GPs with moderate or low recognition rates increases were significant (P = 0.001). Conversation on psychological topics increased by 3.2% (95% CI -0.7% to 7.1%), and by 7.0% (95% CI 1.8% to 12.3%) for patients screened positive. Rates of planned follow-up consultations increased by 3.9% (95% CI 0.6% to 7.3%) and by 4.9% (95% CI 0.7% to 9.1%) for patients screened positive. GPs' self-reported benefit from screening was related to better outcomes. A range of patient and GP factors suggesting added value from using SQs were identified. CONCLUSION: GPs' recognition and provision of mental health care can be influenced by the use of composite SQs. Perceived benefit from screening may serve as a useful predictor of better patient management. Pragmatic case-finding approaches need further evaluation. PMID- 15897213 TI - Cough mixtures: rational or irrational prescribing in Hong Kong? AB - OBJECTIVES: To investigate the extent and how cough mixtures are prescribed, and what conditions or specific groups of people would contribute to its prescription in Hong Kong. METHODS: Using diagnosis and drug data obtained from logbooks submitted by participants in the diploma in family medicine course between 1999 and 2003, we selected and analysed all patients with a diagnosis of cough or cough-related illnesses as well as cough mixtures that were used to treat them. RESULTS: This study confirmed that cough-related illnesses were common in the Hong Kong primary care setting and cough mixtures were used quite liberally irrespective of the patients' age and sex. Combination preparations accounted for over half of the prescriptions and cough mixture was used less in severe cases when antibiotics were given. Private doctors working in the public sector. CONCLUSION: Given the current health care system, inappropriate and over prescribing of cough mixtures can be improved by promoting health education and awareness among patients seeking medical help for this common medical condition. PMID- 15897214 TI - Electronic primary care guidelines with links to Cochrane reviews--EBM Guidelines. AB - In Finland, guidelines have been used in primary care since the late 1980s to bridge the gap between research evidence and practice. From the very beginning, the electronic format has been the primary source for EBM Guidelines (EBMG). Although also published on CD and in print, the guidelines are currently mostly used via the Internet. Keeping more than 1000 guidelines up to date is a great challenge to the editorial group, and several methods are used to guarantee the quality. The Cochrane Library has become the most important source of information for the guideline producers and all Cochrane reviews relevant to GPs are summarized and linked to the guidelines. In this article we present our experience of producing the electronic guidelines for GPs, the methodology used, and data on the clinical use of these guidelines and their Cochrane links. We also discuss the barriers and facilitators to our process and present ideas for future development. PMID- 15897215 TI - The integral role of non-clinical academics in meeting the goals of primary care training and research. PMID- 15897216 TI - Worsening myopathy associated with ezetimibe in a patient with McArdle disease. PMID- 15897217 TI - Discordant public and professional perceptions on transparency in healthcare. PMID- 15897218 TI - Post-traumatic hyponatraemia due to acute hypopituitarism. PMID- 15897219 TI - Anna O and the 'talking cure'. PMID- 15897220 TI - Proposal of a comprehensive clinical typology of alcohol withdrawal--a cluster analysis approach. AB - AIMS: To characterize the various courses of alcohol withdrawal. METHODS: The Alcohol Withdrawal Scale (AWS) was applied to 217 alcohol-dependent patients every 4 h till the symptoms of withdrawal had passed (until each of four consecutive scores were <3). Patients were medicated by a standardized treatment scheme according to AWS-scores. Hierarchical cluster analysis and discriminant analysis were applied. RESULTS: We found five clusters representing increasing severity of alcohol withdrawal. Each cluster is characterized by a combination of the two maximum subscores (vegetative and psychopathological subscore) and three additional psychopathological symptoms (anxiety, disorientation, and hallucination). In 18.4% of the patients, relevant symptoms were not observed (cluster 1), 18.9% developed mild or moderate vegetative symptoms only (cluster 2), and 40.6% additional anxiety (cluster 3). In cluster 4 (11.1%) the most frequent psychopathological symptoms were disorientation and anxiety but no hallucinations, which could be observed only in cluster 5 (11.1%). Discriminant analysis using the maximum subscores at the first day of treatment as independent variables correctly predicted 89.9% of the five clusters. CONCLUSIONS: Our findings support a model of alcohol withdrawal clustering along the two dimensions of vegetative and psychopathological severity. Furthermore, the AWS may be useful to predict the course of alcohol withdrawal already at the first day of treatment. PMID- 15897221 TI - Effects of naltrexone on the ethanol-induced changes in the rat central dopaminergic system. AB - AIMS: The opioid antagonist naltrexone may reduce ethanol reward, but the underlying neurochemical mechanisms has yet to be clarified. The afferent projections to the nucleus accumbens from the ventral tegmental area (VTA) provide a potential substrate by which endogenous opioids may modulate the dopaminergic rewarding effects of ethanol. We assessed mRNA levels of tyrosine hydroxylase (TH), a major regulatory enzyme in the dopamine synthesis and levels of dopamine and its metabolites after chronic ethanol administration with and without concomitant naltrexone. METHODS: Sprague-Dawley rats were exposed to chronic ethanol consumption (5%, 4 weeks) with and without concomitant naltrexone administration. Levels of TH mRNA in the VTA and substantia nigra (SN) and dopamine and its metabolites in the striatum of the rats were measured by in situ hybridization and by high performance liquid chromatography, respectively. RESULTS: Chronic ethanol consumption increased TH mRNA levels in the VTA, but did not cause any significant change in the SN. With naltrexone treatment, ethanol induced increase in the TH mRNA level was reduced in the VTA. Chronic ethanol consumption did not cause any change in the levels of dopamine and its metabolites in most brain regions. Only in the striatum, ethanol consumption with naltrexone treatment significantly increases the dopamine level. CONCLUSION: This finding supports the presence of interactions of opioid and dopaminergic systems in the VTA in mediating ethanol reward, and thus naltrexone attenuates the rewarding properties of ethanol by interfering with the ethanol-induced stimulation of the mesolimbic dopaminergic pathway. PMID- 15897222 TI - The genetic background for streptomycin resistance in Escherichia coli influences the distribution of MICs. AB - OBJECTIVES: The aim of this study was to investigate the genetic background for streptomycin resistance in Escherichia coli and perform analysis of the MICs in relation to genetic background. METHODS: The 136 strains investigated, with streptomycin MICs of > or =16 mg/L, originated from meat and meat products and were collected within the frame of the Norwegian monitoring programme for antimicrobial resistance in bacteria from feed, food and animals (NORM-VET). PCR was carried out for detection of the streptomycin resistance genes strA-strB and the integron-associated aadA gene cassettes. RESULTS: The strA-strB genes and/or an aadA gene cassette were detected in 110 of the 136 (80.9%) strains investigated. The strA-strB genes were the most prevalent, and were detected in 90 strains. The aadA gene cassettes were detected in 29 strains, and nine strains harboured both the strA-strB genes and an aadA gene cassette. The distribution of MICs differed considerably between isolates harbouring the strA-strB genes (solely) (MIC(50) = 128 mg/L) and isolates harbouring an aadA gene cassette (solely) (MIC(50) = 16 mg/L). Strains harbouring both the strA-strB genes and an aadA gene cassette had higher streptomycin MICs than those harbouring either alone. CONCLUSIONS: The distribution of streptomycin MICs in E. coli can be greatly influenced by the genes encoding resistance to streptomycin. The strA strB genes are probably involved in conferring high-level resistance to streptomycin, whereas the opposite seems to be the case for the aadA gene cassettes. The low-level streptomycin resistance, caused by the presence of aadA gene cassettes in integrons, represents an obstacle in classifying E. coli as susceptible or resistant to streptomycin. Furthermore, the determination of an epidemiological cut-off value for surveillance purposes is also complicated by dissemination of integrons containing the aadA cassettes. PMID- 15897223 TI - Reduction in outpatient antibiotic sales for pre-school children: interrupted time series analysis of weekly antibiotic sales data in Sweden 1992-2002. AB - OBJECTIVES: The aim of this study was to use detailed weekly data on outpatient antibiotic sales for pre-school children in Sweden to test for the significance of trends during 1992-2002. We also report on the special features found in weekly antibiotic data, and how the interrupted time series (ITS) design can adjust for this. METHODS: Weekly data on the total number of dispensed outpatient antibiotic prescriptions to pre-school children were studied, as well as the individual subgroups commonly used to treat respiratory tract infections in children: narrow-spectrum penicillins, broad-spectrum penicillins and macrolides. In parallel, monthly data of paracetamol sales of paediatric dosages were analysed to reflect trends in symptomatic treatment. An ITS model controlling for seasonality and autocorrelation was used to examine the datasets for significant level and trend shifts. RESULTS: A significant increase in mean and change in level could be found in the total antibiotic data in 1997, also reflected in broad-spectrum penicillin data where a similar trend break occurred in 1996. For macrolides, a trend break with a decrease in mean was noted in 1996, but no trend breaks were found in narrow-spectrum penicillin data. In contrast to the general decreasing trends in antibiotic sales, the yearly over-the-counter sales of paracetamol in paediatric preparations increased during the same period, with no identified trend breaks. CONCLUSIONS: The overall decrease in antibiotic sales and increase in paediatric paracetamol sales might suggest that symptomatic treatment in the home has increased, as antibiotics are less commonly prescribed. PMID- 15897225 TI - Concerning: 'Mobile phones and driving'. PMID- 15897226 TI - Estimation of leaf water potential by thermal imagery and spatial analysis. AB - Canopy temperature has long been recognized as an indicator of plant water status and as a potential tool for irrigation scheduling. In the present study, the potential of using thermal images for an in-field estimation of the water status of cotton under a range of irrigation regimes was investigated. Thermal images were taken with a radiometric infrared video camera. Specific leaves that appeared in the camera field of view were sampled, their LWP was measured and their temperature was calculated from the images. Regression models were built in order to predict LWP according to the crop canopy temperature and to the empirical formulation of the crop water stress index (CWSI). Statistical analysis revealed that the relationship between CWSI and LWP was more stable and had slightly higher correlation coefficients than that between canopy temperature and LWP. The regression models of LWP against CWSI and against leaf temperatures were used to create LWP maps. The classified LWP maps showed that there was spatial variability in each treatment, some of which may be attributed to the difference between sunlit and shaded leaves. The distribution of LWP in the maps showed that irrigation treatments were better distinguished from each other when the maps were calculated from CWSI than from leaf temperature alone. Furthermore, the inclusion of the spatial pattern in the classification enhanced the differences between the treatments and was better matched to irrigation amounts. Optimal determination of the water status from thermal images should be based on an overall view of the physical status as well as on the analysis of the spatial structure. Future study will involve investigating the robustness of the models and the potential of using water status maps, derived from aerial thermal images, for irrigation scheduling and variable management in commercial fields. PMID- 15897227 TI - Dynamics of aquaporins and water relations during hypocotyl elongation in Ricinus communis L. seedlings. AB - The rate of water flow across biological membranes can be modulated by aquaporins which are expressed in many cells and tissues. The biological functions of these water channels in cellular processes have often been anticipated from the expression pattern, although the participation in the underlying process is not known in many cases. Ten putative aquaporin transcripts were identified in castor bean (Ricinus communis L.) seedlings and the water channel activity of three selected genes was analysed by heterologous expression in Xenopus oocytes, as well as the spatial and temporal expression by in situ hybridization/immunolocalization along the hypocotyl's axis. Water relations parameters were studied in elongating and non-elongating tissues using the cell pressure probe technique. These results indicate that (i) the amount of the RcPIP2-1 aquaporin correlated best with the elongation activity of the etiolated hypocotyl and (ii) the hydraulic conductivity of cortex cells is significantly higher in the elongating region of the hypocotyl compared with the non elongating, mature region. PMID- 15897228 TI - The strawberry gene Cyf1 encodes a phytocystatin with antifungal properties. AB - An EST, encoding a strawberry phytocystatin (PhyCys) obtained from a developing fruit of Fragariaxananassa cv. Elsanta has been characterized. The corresponding gene (Cyf1) had three introns interrupting its ORF that codes for a protein (FaCPI-1) of 235 amino acid residues with a putative signal peptide of 29 residues and an estimated molecular mass for the mature protein of 23.1 kDa. This protein contains, besides a C-terminal extension, several motifs conserved in all members of the PhyCys superfamily: (i) a GG and LARFAV-like motifs towards the N terminal part of the protein; (ii) the reactive site QVVAG, and (iii) a conserved PW, downstream of the reactive site. Northern blot and in situ hybridization analyses indicated that the Cyf1 gene was expressed in fully expanded leaves, in roots and in achenes, but not in the receptacle (pseudocarp) during fruit development. The recombinant FaCPI-1 protein expressed in E. coli efficiently inhibited papain (K(i) 1.9 x 10(-9) M) and less so cathepsin H (K(i) 4.7 x 10(-7) M) and cathepsin B (K(i) 3.3 x 10(-6) M), and was a good inhibitor of the in vitro growth of phytopathogenic fungi Botrytis cinerea (EC(50): 1.90 microM) and Fusarium oxysporum (EC(50): 2.28 microM). PMID- 15897229 TI - Phytosulphokine gene regulation during maize (Zea mays L.) reproduction. AB - The sulphated pentapeptide phytosulphokine (PSK) was identified as a substance that promotes cell division in low-density suspension cultures and has been implicated in various aspects of tissue differentiation in plants. The peptide is derived from PSK precursor proteins that are encoded by small gene families. The physiological roles of PSK are still not clearly defined and little is known about expression of members of the PSK precursor gene family in any plant species. In this study, highly regulated tissue and cell type-specific expression are described for four PSK genes from maize (Zea mays L.) in female and male gametophytes, and during seed development. ZmPSK1 and ZmPSK3 were specifically and differentially expressed in cells of female and male gametophytes and in female and male gametes. In anthers ZmPSK1 or ZmPSK3 transcripts were found, for example, at high levels in secretory tapetal cells which support developing microspores. ZmPSK1 mRNA was abundant in mature pollen including sperm cells. ZmPSK1 and ZmPSK3 transcripts were also detected in egg and central cells of the female gametophyte and ZmPSK1 mRNA was present in synergids, indicating that the PSK peptide probably plays a role during gametogenesis, pollen germination, and fertilization. In developing maize kernels all four ZmPSK genes were expressed, albeit with striking differences in their expression patterns. It is proposed here that PSK is required for numerous but defined processes during gametophyte and early sporophyte development. In general, PSK availability appears to be controlled through transcriptional regulation in a tissue and cell type-specific and development-dependent manner. PMID- 15897230 TI - Functional characterization of the thi1 promoter region from Arabidopsis thaliana. AB - The Arabidopsis thaliana THI1 protein is involved in thiamine biosynthesis and is targeted to both chloroplasts and mitochondria by N-terminal control regions. To investigate thi1 expression, a series of thi1 promoter deletions were fused to the beta-glucuronidase (GUS) reporter gene. Transgenic plants were generated and expression patterns obtained under different environmental conditions. The results show that expression derived from the thi1 promoter is detected early on during development and continues throughout the plant's life cycle. High levels of GUS expression are observed in both shoots and roots during vegetative growth although, in roots, expression is restricted to the vascular system. Deletion analysis of the thi1 promoter region identified a region that is responsive to light. The smallest fragment (designated Pthi322) encompasses 306 bp and possesses all the essential signals for tissue specificity, as well as responsiveness to stress conditions such as sugar deprivation, high salinity, and hypoxia. PMID- 15897231 TI - Functional conservation of Notch1 and Notch2 intracellular domains. AB - The Notch receptor is a key component of a highly conserved signaling pathway that regulates cell fate determination during development. In Drosophila, where Notch signaling was first identified and studied, there is only one Notch receptor. In contrast, mammals have four Notch receptor genes, Notch1-4. Notch1 and Notch2 are both required for embryo viability, are widely expressed in mammals, and are structurally conserved. It is presently unknown if these two receptors are functionally redundant or if they have unique capabilities related to differences in their amino acid sequences. In contrast to the rest of the molecule, the amino acid sequences of a large region of the Notch intracellular domain are not highly conserved and thus may be able to interact with distinct transcription factors and mediate the expression of different sets of genes. To determine if the function of this region is conserved, the last 426 amino acids of the Notch2 receptor have been replaced with the corresponding region of Notch1 in mice by using gene targeting. We have determined that even though the amino acid sequences of this region are only 37% identical (137/426), the C-terminal region of the Notch1 intracellular domain can functionally replace that of Notch2 in vivo. PMID- 15897232 TI - Initial apoptosis is followed by increased proliferation of apoptosis-resistant endothelial cells. AB - We have demonstrated that VEGF receptor blockade in combination with chronic hypoxia causes in rats severe angioproliferative pulmonary hypertension (SAPH) associated with arterial occlusion by proliferating endothelial cells, and we postulate that the established, lumen-occluding lesions are the result of the emergence of apoptosis-resistant proliferating cells. To study the dependence of exuberant endothelial cell proliferation on initial apoptosis, we adapted the CELLMAX artificial capillary system to analyze the effects of a VEGF receptor antagonist (SU5416) on human pulmonary microvascular endothelial cells under pulsatile shear stress. Immunohistochemical staining for caspase-3 and PCNA and flow cytometry for Annexin-V and BrdU supported our concept, since SU5416 caused initial apoptosis (35.8% at 24 h after the SU5416 addition and 4.8% in control cells) whereas the surviving cells became hyperproliferative (PCNA positive). Flow cytometry showed that apoptosis inhibition prevented the proliferation following the initial apoptosis. These lumen-filling endothelial cells were apoptosis resistant, grew without serum, and were phenotypically altered in that they express the tumor marker survivin. Hyperproliferative apoptosis-resistant cells were also generated by adding apoptosed cells instead of the VEGF receptor blocker to the CELLMAX system. In conclusion, endothelial cell death resulted in the selection of an apoptosis-resistant, proliferating phenotypically altered endothelial cell phenotype. PMID- 15897233 TI - The anti-invasive activity of cyclooxygenase inhibitors is regulated by the transcription factor ATF3 (activating transcription factor 3). AB - We previously showed that nonsteroidal anti-inflammatory drugs (NSAID) such as sulindac sulfide, which has chemopreventive activity, modulate the expression of several genes detected by microarray analysis. Activating transcription factor 3 (ATF3) was selected for further study because it is a transcription factor involved in cell proliferation, apoptosis, and invasion, and its expression is repressed in human colorectal tumors as compared with normal adjacent tissue. In this report, we show that ATF3 mRNA and protein expression are up-regulated in HCT-116 human colorectal cancer cells following treatment with NSAIDs, troglitazone, diallyl disulfide, and resveratrol. To ascertain the biological significance of ATF3, we overexpressed full-length ATF3 protein in the sense and antisense orientations. Overexpression of ATF3 in the sense orientation decreased focus formation in vitro and reduced the size of mouse tumor xenografts by 54% in vivo. Conversely, overexpression of antisense ATF3 was protumorigenic in vitro, however, not in vivo. ATF3 in the sense orientation did not modulate apoptosis, indicating another mechanism is involved. With microarray analysis, several genes relating to invasion and metastasis were identified by ATF3 overexpression and were confirmed by real-time reverse transcription-PCR, and several of these genes were modulated by sulindac sulfide, which inhibited invasion in these cells. Furthermore, overexpression of ATF3 inhibited invasion to a similar degree as sulindac sulfide treatment, whereas antisense ATF3 increased invasion. In conclusion, ATF3 represents a novel mechanism in which NSAIDs exert their anti invasive activity, thereby linking ATF3 and its gene regulatory activity to the biological activity of these compounds. PMID- 15897234 TI - N-glycosylation and microtubule integrity are involved in apical targeting of prostate-specific membrane antigen: implications for immunotherapy. AB - Prostate-specific membrane antigen (PSMA) is an important biomarker expressed in prostate cancer cells with levels proportional to tumor grade. The membrane association and correlation with disease stage portend a promising role for PSMA as an antigenic target for antibody-based therapies. Successful application of such modalities necessitates a detailed knowledge of the subcellular localization and trafficking of target antigen. In this study, we show that PSMA is expressed predominantly in the apical plasma membrane in epithelial cells of the prostate gland and in well-differentiated Madin-Darby canine kidney cells. We show that PSMA is targeted directly to the apical surface and that sorting into appropriate post-Golgi vesicles is dependent upon N-glycosylation of the protein. Integrity of the microtubule cytoskeleton is also essential for delivery and retention of PSMA at the apical plasma membrane domain, as destabilization of microtubules with nocodazole or commonly used chemotherapeutic Vinca alkaloids resulted in the basolateral expression of PSMA and increased the uptake of anti-PSMA antibody from the basolateral domain. These results may have important relevance to PSMA based immunotherapy and imaging strategies, as prostate cancer cells can maintain a well-differentiated morphology even after metastasis to distal sites. In contrast to antigens on the basolateral surface, apical antigens are separated from the circulation by tight junctions that restrict transport of molecules across the epithelium. Thus, antigens expressed on the apical plasma membrane are not exposed to intravenously administered agents. The ability to reverse the polarity of PSMA from apical to basolateral could have significant implications for the use of PSMA as a therapeutic target. PMID- 15897235 TI - Circulating plasma vascular endothelial growth factor in mice bearing human ovarian carcinoma xenograft correlates with tumor progression and response to therapy. AB - Vascular endothelial growth factor (VEGF) performs as an angiogenic and permeability factor in ovarian cancer, and its overexpression has been associated with poor prognosis. However, models to study its role as a marker of tumor progression are lacking. We generated xenograft variants derived from the A2780 human ovarian carcinoma (1A9), stably transfected with VEGF(121) in sense (1A9-VS 1) and antisense orientation (1A9-VAS-3). 1A9, 1A9-VS-1, and 1A9-VAS-3 disseminated in the peritoneal cavity of nude mice, but only 1A9-VS-1, the VEGF(121)-overexpressing tumor variant, produced ascites. Tumor biopsies from 1A9 VS-1 showed alterations in the vascular pattern and caused an angiogenic response in the chorioallantoic membrane assay. A significant level of soluble VEGF was detectable in the plasma of mice bearing 1A9-VS-1 even at an early stage of tumor growth. Plasma VEGF correlated positively with tumor burden in the peritoneal cavity and ascites accumulation. Cisplatin reduced the tumor burden and ascites in mice bearing 1A9-VS-1; the response was associated with a significant decrease of VEGF in plasma. This 1A9-VS-1 xenograft model reproduces the behavior of human ovarian cancer by growing in the peritoneal cavity, being highly malignant, and producing ascites. Plasma VEGF as a marker of tumor progression offers a valuable means of detecting early tumor response and following up treatments in an animal model. PMID- 15897236 TI - Protein kinase C-{alpha} mediates epidermal growth factor receptor transactivation in human prostate cancer cells. AB - Progression of human prostate cancer to a malignancy that is refractory to androgen-ablation therapy renders the disease resistant to available treatment options and accounts for the high prostate cancer mortality rate. Epidermal growth factor receptor (EGFR) expression in human prostate cancer specimens increases with disease progression to androgen-refractory prostate cancer, and experimental models implicate EGFR-dependent signaling to Erk1/2 activation in the androgen-refractory prostate cancer phenotype. 12-O-Tetradecanoylphorbol-13 acetate (TPA)-induced Erk1/2 activation in human prostate cancer PC-3 cells is a paradigm of diacylglycerol-induced EGFR transactivation in androgen-independent prostate cancer. In this report, we establish an obligatory role for TPA-induced protein kinase C (PKC)-alpha activation in EGFR transactivation and signaling to Erk1/2 activation in PC-3 cells. TPA-regulated molecules include PKCs, PKDs, and Ras guanyl nucleotide-releasing proteins. The PKC-selective inhibitors GF109203X and Go6983 each blocked TPA-induced EGFR transactivation, indicating a requirement for PKC. PC-3 cells express four PKC isozymes. Prolonged bryostatin 1 treatment abrogated PKCalpha expression without altering expression levels of the other PKC isozymes. Pharmacologic PKCalpha "knockdown" abrogated TPA-induced Erk1/2 activation without affecting the EGF/EGFR-induced response, indicating that PKCalpha was required for EGFR transactivation but dispensable for signaling of ligand-activated EGFR to Erk1/2 activation. We corroborated this by showing that Go6976, which is a PKCalpha-selective inhibitor in PC-3 cells, likewise abolished TPA-induced Erk1/2 activation and did not inhibit EGF/EGFR-induced Erk1/2 activation. Go6976 had similar effects in DU145 cells, providing evidence for a common PKCalpha-dependent Erk1/2 activation mechanism in androgen independent human prostate cancer cells of distinct genetic origin. These results constitute a rational basis for selective PKCalpha inhibition as a modality of prostate cancer therapy. PMID- 15897237 TI - Involvement of cathepsin D in chemotherapy-induced cytochrome c release, caspase activation, and cell death. AB - Treatment of cells with chemotherapy drugs activates the intrinsic mitochondrial pathway of apoptosis and the caspase protease cascade. Recently, the lysosomal protease cathepsin D has been implicated in apoptosis caused by oxidative stress, inhibition of protein kinase C, and stimulation of the TNFR1 and Fas death receptors. However, the role of cathepsin D in chemotherapy-induced cell death has remained largely unexplored. In this report, we show that treatment of U937 leukemia cells with the chemotherapy drug etoposide (VP-16) results in cathepsin D release into the cytosol within 4 hours after initiation of drug treatment. VP 16-induced cathepsin D release was not inhibited by z-VAD-FMK or pepstatin A, suggesting that it occurred independently of the activities of caspase proteases or cathepsin D. Down-regulation of cathepsin D expression in suspension U937 cells or adherent HeLa cells using cathepsin D small interfering RNA partially inhibited cell death resulting from treatment of cells with tumor necrosis factor alpha, tumor necrosis factor-related apoptosis inducing ligand, or the chemotherapy drugs VP-16, cisplatin, and 5-fluorouracil. Moreover, cathepsin D down-regulation significantly delayed cytochrome c release and caspase-3 activation in response to chemotherapy treatment. Incubation of isolated mitochondria with cathepsin D-treated cytosolic extracts resulted in potent release of cytochrome c, indicating that a cytoplasmic substrate mediates the effects of cathepsin D on mitochondria. Together, these findings show that cathepsin D plays an important role in chemotherapy-induced cell death, and that cathepsin D lies upstream of cytochrome c release and caspase-3 activation in the chemotherapy-induced execution pathway. PMID- 15897238 TI - Overexpression of hRFI (human ring finger homologous to inhibitor of apoptosis protein type) inhibits death receptor-mediated apoptosis in colorectal cancer cells. AB - The acquisition of antiapoptotic properties is one of the essential mechanistic steps in colorectal carcinogenesis and is closely correlated with a loss of chemosensitivity and radiosensitivity. Human ring finger homologous to inhibitor of apoptosis protein type (hRFI) is a newly discovered gene encoding a ring finger domain highly homologous to that of X chromosome-linked inhibitor of apoptosis protein. Immunohistochemistry has revealed that the expression of hRFI increased in transition from normal colorectal mucosas to adenomas and from adenomas to carcinomas, suggesting an essential role in the early stage of colorectal carcinogenesis. However, the function role of hRFI in colorectal carcinoma has not been elucidated. To determine whether hRFI possesses an antiapoptotic function in colorectal cancer cells, HCT116 colorectal cancer cells stably overexpressing hRFI were established. The hRFI transfectant exhibited significant resistance to apoptosis induced by tumor necrosis factor-alpha or tumor necrosis factor-related apoptosis-inducing ligand compared with control. This antiapoptotic response was associated with decreased activity of caspase-3, 8, and -9. We also established an antisense down-regulation of hRFI, which effectively reversed the antiapoptotic activity of the hRFI transfectant. This confirmed that the antiapoptotic property of the hRFI transfectant was not due to the clonal effect but in fact dependent on hRFI function. In conclusion, hRFI possesses an antiapoptotic function in HCT116 colorectal cancer cells. Considering the progressive increase of hRFI expression in the advance of the colorectal adenoma-carcinoma sequence, hRFI is one of the important players in colorectal carcinogenesis through its effect on apoptosis regulation. PMID- 15897239 TI - Matrix metalloproteinase-activated doxorubicin prodrugs inhibit HT1080 xenograft growth better than doxorubicin with less toxicity. AB - Matrix metalloproteinase (MMP)-activated prodrugs were formed by coupling MMP cleavable peptides to doxorubicin. The resulting conjugates were excellent in vitro substrates for MMP-2, -9, and -14. HT1080, a fibrosarcoma cell line, was used as a model system to test these prodrugs because these cells, like tumor stromal fibroblasts, expressed several MMPs. In cultured HT1080 cells, simple MMP cleavable peptides were primarily metabolized by neprilysin, a membrane-bound metalloproteinase. MMP-selective metabolism in cultured HT1080 cells was obtained by designing conjugates that were good MMP substrates but poor neprilysin substrates. To determine how conjugates were metabolized in animals, MMP selective conjugates were given to mice with HT1080 xenografts and the distribution of doxorubicin was determined. These studies showed that MMP selective conjugates were preferentially metabolized in HT1080 xenografts, relative to heart and plasma, leading to 10-fold increases in the tumor/heart ratio of doxorubicin. The doxorubicin deposited by a MMP-selective prodrug, compound 6, was more effective than doxorubicin at reducing HT1080 xenograft growth. In particular, compound 6 cured 8 of 10 mice with HT1080 xenografts at doses below the maximum tolerated dose, whereas doxorubicin cured 2 of 20 mice at its maximum tolerated dose. Compound 6 was less toxic than doxorubicin at this efficacious dose because mice treated with compound 6 had no detectable changes in body weight or reticulocytes, a marker for marrow toxicity. Hence, MMP activated doxorubicin prodrugs have a much higher therapeutic index than doxorubicin using HT1080 xenografts as a preclinical model. PMID- 15897240 TI - The discovery and mechanism of action of novel tumor-selective and apoptosis inducing 3,5-diaryl-1,2,4-oxadiazole series using a chemical genetics approach. AB - A novel series of 3,5-diaryl-oxadiazoles was identified as apoptosis-inducing agents through our cell and chemical genetics-based screening assay for compounds that induce apoptosis using a chemical genetics approach. Several analogues from this series including MX-74420 and MX-126374 were further characterized. MX 126374, a lead compound from this series, was shown to induce apoptosis and inhibit cell growth selectively in tumor cells. To elucidate the mechanism(s) by which this class of compounds alters the signal transduction pathway that ultimately leads to apoptosis, expression profiling using the Affymetrix Gene Chip array technology was done along with other molecular and biochemical analyses. Interestingly, we have identified several key genes (cyclin D1, transforming growth factor-beta1, p21, and insulin-like growth factor-BP3) that are altered in the presence of this compound, leading to characterization of the pathway for activation of apoptosis. MX-126374 also showed significant inhibition of tumor growth as a single agent and in combination with paclitaxel in murine tumor models. Using photoaffinity labeling, tail-interacting protein 47, an insulin-like growth factor-II receptor binding protein, was identified as the molecular target. Further studies indicated that down-regulation of tail interacting protein 47 in cancer cells by small interfering RNA shows a similar pathway profile as compound treatment. These data suggest that 3,5-diaryl oxadiazoles may be a new class of anticancer drugs that are tumor-selective and further support the discovery of novel drugs and drug targets using chemical genetic approaches. PMID- 15897241 TI - Strongylophorine-26, a Rho-dependent inhibitor of tumor cell invasion that reduces actin stress fibers and induces nonpolarized lamellipodial extensions. AB - Strongylophorine-26, a new meroditerpenoid, was recently identified as an inhibitor of cancer cell invasion. This study was undertaken to characterize its mechanism of action. We find that strongylophorine-26 inhibits the motility of MDA-MB-231 breast carcinoma cells on a plastic surface. Upon addition of strongylophorine-26, rapid cell contraction and depolarization occurred, followed by spreading and flattening of the entire cell. Treated cells exhibited increased membrane ruffling throughout and extended lamellipodia in all directions. Strongylophorine-26 induced a decrease in actin stress fibers, a dramatic increase in the size and number of focal adhesions, and the appearance of a dense meshwork of actin filaments around the cell periphery. Strongylophorine-26 caused a transient activation of the small GTPase Rho and treatment with the Rho inhibitor C3 exoenzyme abrogated the anti-invasive activity of strongylophorine 26. These effects are distinct from those of many motility and angiogenesis inhibitors that seem to act by a common mechanism involving the induction of actin stress fibers. This difference in mechanism of action sets strongylophorine 26 apart as an experimental anticancer agent and indicates that pharmacologic inhibition of cell migration may be achieved by mechanisms not involving the stabilization of actin stress fibers. PMID- 15897242 TI - Anti-gene peptide nucleic acid specifically inhibits MYCN expression in human neuroblastoma cells leading to cell growth inhibition and apoptosis. AB - We developed an anti-gene peptide nucleic acid (PNA) for selective inhibition of MYCN transcription in neuroblastoma cells, targeted against a unique sequence in the antisense DNA strand of exon 2 of MYCN and linked at its NH(2) terminus to a nuclear localization signal peptide. Fluorescence microscopy showed specific nuclear delivery of the PNA in six human neuroblastoma cell lines: GI-LI-N and IMR-32 (MYCN-amplified/overexpressed); SJ-N-KP and NB-100 (MYCN-unamplified/low expressed); and GI-CA-N and GI-ME-N (MYCN-unamplified/unexpressed). Antiproliferative effects were observable at 24 hours (GI-LI-N, 60%; IMR-32, 70%) and peaked at 72 hours (GI-LI-N, 80%; IMR-32, 90%; SK-N-KP, 60%; NB-100, 50%); no reduction was recorded for GI-CA-N and GI-ME-N (controls). In MYCN amplified/overexpressed IMR-32 cells and MYCN-unamplified/low-expressed SJ-N-KP cells, inhibition was recorded of MYCN mRNA (by real-time PCR) and N-Myc (Western blotting); these inhibitory effects increased over 3 days after single treatment in IMR-32. Anti-gene PNA induced G(1)-phase accumulation (39-53%) in IMR-32 and apoptosis (56% annexin V-positive cells at 24 hours in IMR-32 and 22% annexin V positive cells at 48 hours in SJ-N-KP). Selective activity of the PNA was shown by altering three point mutations, and by the observation that an anti-gene PNA targeted against the noncoding DNA strand did not exert any effect. These findings could encourage research into development of an anti-gene PNA-based tumor-specific agent for neuroblastoma (and other neoplasms) with MYCN expression. PMID- 15897243 TI - Fibroblast growth factor receptor 3 inhibition by short hairpin RNAs leads to apoptosis in multiple myeloma. AB - The presence of t(4;14)(p16.3;q32.3) in multiple myeloma cells results in dysregulated expression of the fibroblast growth factor receptor 3 (FGFR3). FGFR3 acts as an oncogene to promote multiple myeloma cell proliferation and antiapoptosis. These encourage the clinical development of FGFR3-specific inhibitors. Three short hairpin RNAs (shRNA) targeting different sites of FGFR3 were selected and subsequently transfected into KMS-11, OPM-2, and NCI-H929 human myeloma cell lines, all of which are characterized by t(4;14) and FGFR3 over expression. The combination of these three shRNAs can effectively inhibit FGFR3 expression in all three cell lines. Sequential immunocytochemistry/fluorescence in situ hybridization was employed to validate that the shRNAs specifically inhibited FGFR3 expression in OPM-2 cells. Decreased expression of B-cell chronic lymphocytic leukemia/lymphoma 2 (BCL2) and myeloid cell leukemia sequence 1 (MCL1) proteins and increased staining of Annexin V-positive cells showed that inhibition of FGFR3 induces apoptosis. After confirming down-regulation of FGFR3 by real-time PCR, HU-133 plus 2.0 array was employed to compare the gene expression profile of shRNA-treated sample with that of the control. Besides the down-regulation of FGFR3, expression of the antiapoptotic genes CFLAR, BCL2, MCL1, and some members of NF-kappaB family decreased, whereas expression of the proapoptotic genes CYC, BID, CASP2, and CASP6 increased. Microarray results also revealed changes in genes previously implicated in multiple myeloma pathogenesis (RAS, RAF, IL-6R, and VEGF), as well as others (TLR4, KLF4, and GADD45A) not previously linked to multiple myeloma. Our observations indicate that shRNAs can specifically and effectively inhibit FGFR3 expression. This targeted approach may be worth testing in multiple myeloma patients with t(4;14) and FGFR3 overexpression in the future. PMID- 15897244 TI - The role of IFN regulatory factor-3 in the cytotoxic activity of NS-9, a polyinosinic-polycytidylic acid/cationic liposome complex, against tumor cells. AB - NS-9 is a complex of polyinosinic-polycytidylic acid and a novel cationic liposome, LIC-101. The complex has strong cytotoxic activity against tumor cells derived from epithelial or fibroblastic cells. We have investigated the mechanism of the cytotoxic activity of NS-9 using knockdown cells in which the expression of proteins of interest was inhibited by RNA interference. NS-9 showed strong cytotoxic activity against knockdown cells with reduced expression of double stranded RNA-dependent protein kinase, RNase L, or IFN-alpha/beta receptor, but showed no cytotoxic activity against IFN regulatory factor-3 (IRF3) knockdown cells. In IRF3-knockdown cells, NS-9 also did not induce either the DNA fragmentation or the rRNA degradation observed in negative control cells. We conclude that IRF3 plays a crucial role in the cytotoxic activity of NS-9 against tumor cells, whereas RNA-dependent protein kinase, RNase L, or type I IFNs are not important for its activity. PMID- 15897245 TI - Identification of novel targeting peptides for human ovarian cancer cells using "one-bead one-compound" combinatorial libraries. AB - Using "one-bead one-compound" combinatorial chemistry technology, we generated random peptide libraries containing millions of 90 mum TentaGel beads, each with its own unique amino acid sequence. A cyclic random 8-mer library was screened with CAOV-3 (a human ovarian adenocarcinoma cell line) and beads with a unique ligand that bind to the cell surface receptors were coated by one or more layers of cells. These positive beads were isolated, stripped, and microsequenced. Several peptide motifs were identified from these screenings, some of which were novel and unique, e.g., cDGX(4)GX(6)X(7)c. Structure-activity relationship studies of this peptide revealed that the l-aspartate residue at position 2, the two glycines at positions 3 and 5, and the two d-cysteines at the amino and COOH terminus are critical for activity. In addition, a hydrophobic residue was preferred at position X(4), whereas amino acids at positions X(6) and X(7) were more variable. Binding of this peptide to a number of different cancer cell lines and normal cells was also determined and we observed that peptides with this motif bound preferentially to three other human ovarian cancer cell lines (ES-2, SKOV-3, and OVCAR-3) as well as a human glioblastoma cancer cell line (A172). Structural analysis of the peptides using high-resolution nuclear magnetic resonance spectroscopy revealed strong conformational similarity among all peptides with cX(1)GX(4)GX(6)X(7)c motif. Blocking study with a panel of anti integrin antibodies strongly suggests alpha3 integrin present on these ovarian adenocarcinoma cells is the target receptor for this peptide. PMID- 15897246 TI - Transcriptional signature of Ecteinascidin 743 (Yondelis, Trabectedin) in human sarcoma cells explanted from chemo-naive patients. AB - Ecteinascidin 743 (ET-743; Yondelis, Trabectedin) is a marine anticancer agent that induces long-lasting objective remissions and tumor control in a subset of patients with pretreated/resistant soft-tissue sarcoma. Drug-induced tumor control is achievable in 22% of such patients, but there is no clear indication of the molecular features correlated with clinical sensitivity/resistance to ET 743. Nine low-passage, soft-tissue sarcoma cell lines, explanted from chemo-naive patients with different patterns of sensitivity, have been profiled with a cDNA microarray containing 6,700 cancer-related genes. The molecular signature of these cell lines was analyzed at baseline and at four different times after ET 743 exposure. The association of levels of TP53 mutation and TP73 expression with ET-743 sensitivity and cell cycle kinetics after treatment was also analyzed. Gene expression profile analysis revealed up-regulation of 86 genes and down regulation of 244 genes in response to ET-743. The ET-743 gene expression signature identified a group of genes related with cell cycle control, stress, and DNA-damage response (JUNB, ATF3, CS-1, SAT, GADD45B, and ID2) that were up regulated in all the cell lines studied. The transcriptional signature 72 hours after ET-743 administration, associated with ET-743 sensitivity, showed a more efficient induction of genes involved in DNA-damage response and apoptosis, such as RAD17, BRCA1, PAR4, CDKN1A, and P53DINP1, in the sensitive cell line group. The transcriptional signature described here may lead to the identification of ET 743 downstream mediators and transcription regulators and the proposal of strategies by which ET-743-sensitive tumors may be identified. PMID- 15897247 TI - The selective retinoid X receptor agonist bexarotene (LGD1069, Targretin) prevents and overcomes multidrug resistance in advanced breast carcinoma. AB - Acquired drug resistance represents a major challenge in the therapeutic management of breast cancer patients. We reported previously that the retinoid X receptor-selective agonist bexarotene (LGD1069, Targretin) was efficacious in treating animal models of tamoxifen-resistant breast cancer. The goal of this study was to evaluate the effect of bexarotene on development of acquired drug resistance and its role in overcoming acquired drug resistance in advanced breast cancer. Paclitaxel, doxorubicin, and cisplatin were chosen as model compounds to determine the effect of bexarotene on the development of acquired drug resistance. Human breast cancer cells MDA-MB-231 were repeatedly treated in culture with a given therapeutic agent with or without bexarotene for 3 months. Thereafter, cells were isolated and characterized for their drug sensitivity. Compared with parental cells, cells treated with a single therapeutic agent became resistant to the therapeutic agent, whereas cells treated with the bexarotene combination remained chemosensitive. Cells with acquired drug resistance, when treated with the combination, showed increased sensitivity to the cytotoxic agent. Furthermore, cells treated with the combination regimen had reduced invasiveness and angiogenic potential than their resistant counterparts. These in vitro findings were further confirmed in an in vivo MDA-MB-231 xenograft model. Our results suggest a role for bexarotene in combination with chemotherapeutic agents in prevention and overcoming acquired drug resistance in advanced breast carcinoma. PMID- 15897248 TI - Human mesothelioma cells exhibit tumor cell-specific differences in phosphatidylinositol 3-kinase/AKT activity that predict the efficacy of Onconase. AB - Malignant mesothelioma is an aggressive cancer with no known cure, which has become a therapeutic challenge. Onconase is one of few chemotherapeutic agents that have been studied in patients with malignant mesothelioma that has the advantage of low toxicity and limited side effects. Here, we evaluate the effect of Onconase on killing of malignant mesothelioma cells and how the phosphatidylinositol 3-kinase/AKT (PI3-K/AKT) survival pathway influences this effect. Our results show that Onconase induces apoptosis in malignant mesothelioma cell lines and that this effect is tumor cell specific. Malignant mesothelioma cell lines with the highest AKT activation, which correlated with the presence of the SV40 large and small T antigen (SV40+), were the most resistant to the drug. Finally, a cooperative effect was observed between small molecule inhibitors of PI3-K and Onconase in the killing of malignant mesothelioma cells. Our results suggest that kinase screening of individual malignant mesotheliomas for endogenous levels of activated PI3-K/AKT may be predictive of the efficacy of Onconase and possibly other chemotherapeutic agents. PMID- 15897249 TI - Irinotecan pharmacokinetic and pharmacogenomic alterations induced by methylselenocysteine in human head and neck xenograft tumors. AB - The combination of methylselenocysteine and irinotecan (CPT-11) is synergistic against FaDu and A253 xenografts. Methylselenocysteine/CPT-11 increased tumor cure rate to 100% in FaDu and to 60% in A253. In this study, the effect of methylselenocysteine on pharmacokinetic and pharmacogenetic profiles of genes relevant to CPT-11 metabolic pathway was evaluated to identify possible mechanisms associated with the observed combinational synergy. Nude mice bearing tumors (FaDu and A253) were treated with methylselenocysteine, CPT-11, and a combination of methylselenocysteine/CPT-11. Samples were collected and analyzed for plasma and intratumor concentration of CPT-11 and 7-ethyl-10-hydroxyl camptothecin (SN-38) by high-performance liquid chromatography. The intratumor relative expression of genes related to the CPT-11 metabolic pathway was measured by real-time PCR. After methylselenocysteine treatment, the intratumor area under the concentration-time curve of SN-38 increased to a significantly higher level in A253 than in FaDu and was associated with increased expression of CES1 in both tumors. Methylselenocysteine/CPT-11 treatment, compared with CPT-11 alone, resulted in a significant decrease in levels of ABCC1 and DRG1 in FaDu tumors and an increase in levels of CYP3A5 and TNFSF6 in A253 tumors. No statistically significant changes induced by methylselenocysteine/CPT-11 were observed in the levels of other investigated variables. In conclusion, the significant increase in the cure rate after methylselenocysteine/CPT-11 could be related to increased drug delivery into both tumors (CES1), reduced resistance to SN-38 (ABCC1 and DRG1) in FaDu, and induced Fas ligand apoptosis (TNFSF6) in A253. No correlation was observed between cure rate and other investigated variables (transporters, degradation enzymes, DNA repair, and cell survival/death genes) in either tumor. PMID- 15897250 TI - The multidrug resistance protein 5 (ABCC5) confers resistance to 5-fluorouracil and transports its monophosphorylated metabolites. AB - 5'-Fluorouracil (5-FU), used in the treatment of colon and breast cancers, is converted intracellularly to 5'-fluoro-2'-deoxyuridine (5-FUdR) by thymidine phosphorylase and is subsequently phosphorylated by thymidine kinase to 5'-fluoro 2'-dUMP (5-FdUMP). This active metabolite, along with the reduced folate cofactor, 5,10-methylenetetrahydrofolate, forms a stable inhibitory complex with thymidylate synthase that blocks cellular growth. The present study shows that the ATP-dependent multidrug resistance protein-5 (MRP5, ABCC5) confers resistance to 5-FU by transporting the monophosphate metabolites. MRP5- and vector transfected human embryonic kidney (HEK) cells were employed in these studies. In 3-day cytotoxicity assays, MRP5-transfected cells were approximately 9-fold resistant to 5-FU and 6-thioguanine. Studies with inside-out membrane vesicles prepared from transfected cells showed that MRP5 mediates ATP-dependent transport of 5 micromol/L [(3)H]5-FdUMP, [(3)H]5-FUMP, [(3)H]dUMP, and not [(3)H]5-FUdR, or [(3)H]5-FU. The ATP-dependent transport of 5-FdUMP showed saturation with increasing concentrations and had a K(m) of 1.1 mmol/L and V(max) of 439 pmol/min/mg protein. Uptake of 250 micromol/L 5-FdUMP was inhibited by dUMP, cyclic nucleotide, cyclic guanosine 3',5'-monophosphate, amphiphilic anions such as probenecid, MK571, the phosphodiesterase inhibitors, trequinsin, zaprinast, and sildenafil, and by the chloride channel blockers, 5-nitro-2-(3 phenylpropylamino)-benzoic acid and glybenclamide. Furthermore, the 5-FU drug sensitivity of HEK-MRP5 cells was partially modulated to that of the HEK-vector by the presence of 40 micromol/L 5-nitro-2-(3-phenylpropylamino)-benzoic acid but not by 2 mmol/L probenecid. Thus, MRP5 transports the monophosphorylated metabolite of this nucleoside and when MRP5 is overexpressed in colorectal and breast tumors, it may contribute to 5-FU drug resistance. PMID- 15897251 TI - REM restriction persistently alters strategy used to solve a spatial task. AB - We tested the hypothesis that rapid eye movement (REM) sleep is important for complex associative learning by restricting rats from entering REM sleep for 4 h either immediately after training on an eight-box spatial task (0-4 REMr) or 4 h following training (4-8 REMr). Both groups of REM-restricted rats eventually reached the same overall performance level as did nonrestricted controls, but 0-4 REMr animals were delayed in their improvement in the first few days and lagged behind controls in the middle portion of the training period. More importantly, performance gains of 0-4 REMr rats depended more on simple local cues throughout the 15-d study since, unlike control and 4-8 REMr animals, their error rate increased after daily disruption of the relationship between local (intramaze) cues and the food reward. Thus, although overall performance was only subtly and transiently impaired, due to the ability to use alternate, nonspatial behavioral strategies, complex associative (spatial) learning was persistently impaired by restricting REM for a short critical period each day. PMID- 15897252 TI - Purkinje cell activity in the cerebellar anterior lobe after rabbit eyeblink conditioning. AB - The cerebellar anterior lobe may play a critical role in the execution and proper timing of learned responses. The current study was designed to monitor Purkinje cell activity in the rabbit cerebellar anterior lobe after eyeblink conditioning, and to assess whether Purkinje cells in recording locations may project to the interpositus nucleus. Rabbits were trained in an interstimulus interval discrimination procedure in which one tone signaled a 250-msec conditioned stimulus-unconditioned stimulus (CS-US) interval and a second tone signaled a 750 msec CS-US interval. All rabbits showed conditioned responses to each CS with mean onset and peak latencies that coincided with the CS-US interval. Many anterior lobe Purkinje cells showed significant learning-related activity after eyeblink conditioning to one or both of the CSs. More Purkinje cells responded with inhibition than with excitation to CS presentation. In addition, when the firing patterns of all conditioning-related Purkinje cells were pooled, it appeared that the population showed a pattern of excitation followed by inhibition during the CS-US interval. Using cholera toxin-conjugated horseradish peroxidase, Purkinje cells in recording areas were found to project to the interpositus nucleus. These data support previous studies that have suggested a role for the anterior cerebellar cortex in eyeblink conditioning as well as models of cerebellar-mediated CR timing that postulate that Purkinje cell activity inhibits conditioned response (CR) generation during the early portion of a trial by inhibiting the deep cerebellar nuclei and permits CR generation during the later portion of a trial through disinhibition of the cerebellar nuclei. PMID- 15897253 TI - Fos protein expression in olfactory-related brain areas after learning and after reactivation of a slowly acquired olfactory discrimination task in the rat. AB - Fos protein immunodetection was used to investigate the neuronal activation elicited in some olfactory-related areas after either learning of an olfactory discrimination task or its reactivation 10 d later. Trained rats (T) progressively acquired the association between one odor of a pair and water reward in a four-arm maze. Two groups of pseudotrained rats were used: PO rats were not water restricted and were submitted to the olfactory stimuli in the maze without any reinforcement, whereas PW rats were water-deprived and systematically received water in the maze without any odorous stimulation. When the discrimination task was well mastered, a significantly lower Fos immunoreactivity was observed in T rats compared to PW and PO rats in most of the analyzed brain areas, which could reflect the post-acquisition consolidation process. Following memory reactivation, differences in Fos immunoreactivity between trained and some pseudotrained rats were found in the anterior part of piriform cortex, CA3, and orbitofrontal cortex. We also observed that Fos labeling was significantly higher in trained rats after memory reactivation than after acquisition of the olfactory task in most of the brain areas examined. Our results support the assumption of a differential involvement of neuronal networks after either learning or reactivation of an olfactory discrimination task. PMID- 15897254 TI - Trace fear conditioning is enhanced in mice lacking the delta subunit of the GABAA receptor. AB - The delta subunit of the GABA(A) receptor (GABA(A)R) is highly expressed in the dentate gyrus of the hippocampus. Genetic deletion of this subunit reduces synaptic and extrasynaptic inhibition and decreases sensitivity to neurosteroids. This paper examines the effect of these changes on hippocampus-dependent trace fear conditioning. Compared to controls, delta knockout mice exhibited enhanced acquisition of tone and context fear. Hippocampus-independent delay conditioning was normal in these animals. These results suggest that reduced inhibition in the dentate gyrus facilitates the acquisition of trace fear conditioning. However, the enhancement in trace conditioning was only observed in female knockout mice. The sex-specificity of this effect may be a result of neuroactive steroids. These compounds vary during the estrus cycle, can increase GABAergic inhibition, and have been shown to impair hippocampus-dependent learning. We propose that activation of GABA(A)Rs by neuroactive steroids inhibits learning processes in the hippocampus. Knockouts are immune to this effect because of the reduced neurosteroid sensitivity that accompanies deletion of the delta subunit. Relationships between neurosteroids, hippocampal excitability, and memory are discussed. PMID- 15897255 TI - Working memory deficits in retinoid X receptor gamma-deficient mice. AB - Retinoid signaling has been recently shown to be required for mnemonic functions in rodents. To dissect the behavioral and molecular mechanisms involved in this requirement, we have analyzed the spatial and recognition working memory in mice carrying null mutations of retinoid receptors RARbeta and RXRgamma. Double mutants appeared deficient in spatial working memory as tested in spontaneous alternation in the Y-maze and delayed nonmatch to place (DNMTP) test in the T maze. These mutant mice did acquire, however, spatial place reference or right/left discrimination tasks in the T-maze set-up, indicating that basic sensorimotor functions, spatial orientation, and motivational factors are unlikely to account for deficits in working memory-sensitive tasks. Double-mutant mice were also deficient in novel object recognition at intermediate, but not short delays. RXRgamma appeared to be the functionally predominant receptor in modulation of the working memory, as RXRgamma, but not RARbeta single null mutant mice exhibited deficits similar to those observed in the double mutants. The mechanism of this modulation is potentially related to functions of RXRgamma in frontal and perirhinal cortex, structures in which we detected RXRgamma expression and which are functionally implicated in working memory processes. PMID- 15897256 TI - Trace eyeblink conditioning requires the hippocampus but not autophosphorylation of alphaCaMKII in mice. AB - Little is known about signaling mechanisms underlying temporal associative learning. Here, we show that mice with a targeted point mutation that prevents autophosphorylation of alphaCaMKII (alphaCaMKII(T286A)) learn trace eyeblink conditioning normally. This forms a sharp contrast to the severely impaired spatial learning in the water maze and contextual fear conditioning observed in alphaCaMKII(T286A) mutants. Importantly, hippocampal lesions impaired trace eyeblink conditioning in alphaCaMKII(T286A) mice, suggesting a potential role of hippocampal alphaCaMKII-independent mechanisms. These results indicate that hippocampal signaling mechanisms that underlie temporal associative learning as assessed by trace eyeblink conditioning may differ from those of spatial and contextual learning. PMID- 15897257 TI - Dissociable contributions within the medial temporal lobe to encoding of object location associations. AB - The crucial role of the medial temporal lobe (MTL) in episodic memory is well established. Although there is little doubt that its anatomical subregions-the hippocampus, peri-, entorhinal and parahippocampal cortex (PHC)-contribute differentially to mnemonic processes, their specific functions in episodic memory are under debate. Data from animal, human lesion, and neuroimaging studies suggest somewhat contradictory perspectives on this functional specialization: a general participation in declarative memory, an exclusive involvement in associative mnemonic processes, and a specific contribution to spatial memory are reported for the hippocampus, adjacent cortices, and the PHC. A functional lateralization in humans dependent on the verbalizability of the material is also discussed herein. To further elucidate the differential contributions of the various MTL subregions to encoding, we employed an object-location association memory paradigm. The memory for each of the studied associations was tested twice: by the object, and by the location serving as retrieval cue. The memory accuracy in response to both cue types was also assessed parametrically. Brain activity during encoding which leads to different degrees of subsequent memory accuracy under the two retrieval conditions was compared. We found the bilateral posterior PHC to participate in encoding of both the object associated with a location and the location associated with an object. In contrast, activity in an area in the left anterior PHC and the right anterior MTL was only correlated with the memory for the location associated with an object. PMID- 15897258 TI - Acetylcholine in the orbitofrontal cortex is necessary for the acquisition of a socially transmitted food preference. AB - The social transmission of food preference task (STFP) has been used to examine the involvement of the hippocampus in learning and memory for a natural odor-odor association. However, cortical involvement in STFP has not been extensively studied. The orbitofrontal cortex (OFC) is important in odor-guided learning, and cholinergic depletion of the entire neocortex results in impairments in STFP. Here we examined the specific role of cholinergic modulation in the OFC by assessing the effect of 192 immunoglobulin G-saporin infusion directly into OFC prior to training on STFP. Cholinergic depletion in the OFC impaired expression of the socially transmitted odor association measured 2 d after training, indicating that cholinergic function in the OFC is essential for this form of associative learning. PMID- 15897259 TI - Recollection in an episodic-like memory task in the rat. AB - Episodic memory in humans is the conscious recollection of a past event. Animal models of episodic-like memory assess the memory for "what"happened, "where" it happened, and either "when" it happened, or in "which" context it happened, although recollection on such tasks is often difficult to measure. Here we present the first evidence of successful recollection of a past event in a rat in a task which is easily performed, requires little training, and is easily adaptable for other commonly used laboratory species. PMID- 15897260 TI - Essential function of Drosophila Sec6 in apical exocytosis of epithelial photoreceptor cells. AB - Polarized exocytosis plays a major role in development and cell differentiation but the mechanisms that target exocytosis to specific membrane domains in animal cells are still poorly understood. We characterized Drosophila Sec6, a component of the exocyst complex that is believed to tether secretory vesicles to specific plasma membrane sites. sec6 mutations cause cell lethality and disrupt plasma membrane growth. In developing photoreceptor cells (PRCs), Sec6 but not Sec5 or Sec8 shows accumulation at adherens junctions. In late PRCs, Sec6, Sec5, and Sec8 colocalize at the rhabdomere, the light sensing subdomain of the apical membrane. PRCs with reduced Sec6 function accumulate secretory vesicles and fail to transport proteins to the rhabdomere, but show normal localization of proteins to the apical stalk membrane and the basolateral membrane. Furthermore, we show that Rab11 forms a complex with Sec5 and that Sec5 interacts with Sec6 suggesting that the exocyst is a Rab11 effector that facilitates protein transport to the apical rhabdomere in Drosophila PRCs. PMID- 15897261 TI - Endoplasmic reticulum stress signaling transmitted by ATF6 mediates apoptosis during muscle development. AB - Although apoptosis occurs during myogenesis, its mechanism of initiation remains unknown. In a culture model, we demonstrate activation of caspase-12, the initiator of the endoplasmic reticulum (ER) stress-specific caspase cascade, during apoptosis associated with myoblast differentiation. Induction of ER stress responsive proteins (BiP and CHOP) was also observed in both apoptotic and differentiating cells. ATF6, but not other ER stress sensors, was specifically activated during apoptosis in myoblasts, suggesting that partial but selective activation of ER stress signaling was sufficient for induction of apoptosis. Activation of caspase-12 was also detected in developing muscle of mouse embryos and gradually disappeared later. CHOP was also transiently induced. These results suggest that specific ER stress signaling transmitted by ATF6 leads to naturally occurring apoptosis during muscle development. PMID- 15897262 TI - Histone modifications affect timing of oligodendrocyte progenitor differentiation in the developing rat brain. AB - Timely differentiation of progenitor cells is critical for development. In this study we asked whether global epigenetic mechanisms regulate timing of progenitor cell differentiation into myelin-forming oligodendrocytes in vivo. Histone deacetylation was essential during a specific temporal window of development and was dependent on the enzymatic activity of histone deacetylases, whose expression was detected in the developing corpus callosum. During the first 10 postnatal days, administration of valproic acid (VPA), the specific inhibitor for histone deacetylase activity, resulted in significant hypomyelination with delayed expression of late differentiation markers and retained expression of progenitor markers. Differentiation resumed in VPA-injected rats if a recovery period was allowed. Administration of VPA after myelination onset had no effect on myelin gene expression and was consistent with changes of nucleosomal histones from reversible deacetylation to more stable methylation and chromatin compaction. Together, these data identify global modifications of nucleosomal histones critical for timing of oligodendrocyte differentiation and myelination in the developing corpus callosum. PMID- 15897263 TI - Inducible ablation of mouse Langerhans cells diminishes but fails to abrogate contact hypersensitivity. AB - Langerhans cells (LC) form a unique subset of dendritic cells (DC) in the epidermis but so far their in vivo functions in skin immunity and tolerance could not be determined, in particular in relation to dermal DC (dDC). Here, we exploit a novel diphtheria toxin (DT) receptor (DTR)/DT-based system to achieve inducible ablation of LC without affecting the skin environment. Within 24 h after intra peritoneal injection of DT into Langerin-DTR mice LC are completely depleted from the epidermis and only begin to return 4 wk later. LC deletion occurs by apoptosis in the absence of inflammation and, in particular, the dDC compartment is not affected. In LC-depleted mice contact hypersensitivity (CHS) responses are significantly decreased, although ear swelling still occurs indicating that dDC can mediate CHS when necessary. Our results establish Langerin-DTR mice as a unique tool to study LC function in the steady state and to explore their relative importance compared with dDC in orchestrating skin immunity and tolerance. PMID- 15897264 TI - Palmitoylation regulates plasma membrane-nuclear shuttling of R7BP, a novel membrane anchor for the RGS7 family. AB - The RGS7 (R7) family of RGS proteins bound to the divergent Gbeta subunit Gbeta5 is a crucial regulator of G protein-coupled receptor (GPCR) signaling in the visual and nervous systems. Here, we identify R7BP, a novel neuronally expressed protein that binds R7-Gbeta5 complexes and shuttles them between the plasma membrane and nucleus. Regional expression of R7BP, Gbeta5, and R7 isoforms in brain is highly coincident. R7BP is palmitoylated near its COOH terminus, which targets the protein to the plasma membrane. Depalmitoylation of R7BP translocates R7BP-R7-Gbeta5 complexes from the plasma membrane to the nucleus. Compared with nonpalmitoylated R7BP, palmitoylated R7BP greatly augments the ability of RGS7 to attenuate GPCR-mediated G protein-regulated inward rectifying potassium channel activation. Thus, by controlling plasma membrane nuclear-shuttling of R7BP-R7 Gbeta5 complexes, reversible palmitoylation of R7BP provides a novel mechanism that regulates GPCR signaling and potentially transduces signals directly from the plasma membrane to the nucleus. PMID- 15897265 TI - Risk factors for adverse drug events: a 10-year analysis. AB - BACKGROUND: Many adverse drug events (ADEs) are the result of known pharmacologic properties, and some result from medication errors. However, some are the result of patient-specific risk factors. OBJECTIVE: To identify inpatient risk factors for ADEs. METHODS: Conditional logistic regression was used to analyze all pharmacist-verified ADEs by therapeutic class of drugs and severity during a 10 year study period. All inpatients > or = 18 years of age from a 520-bed tertiary teaching hospital were included. Each case patient was matched with up to 16 control patients. Odds ratios for patient factors associated with ADEs were calculated from different therapeutic classes of drugs. RESULTS: Odds ratios for numerous risk factors were identified for 4376 ADEs and were found to vary depending on therapeutic classification. The risk factors for the different classifications were grouped by (1) patient characteristics--female (OR 1.5-1.7), age (0.7-0.9), weight (1.2-1.4), creatinine clearance (0.8-4.7), and number of comorbidities (1.1-12.6); (2) drug administration--dosage (1.2-3.7), administration route (1.4-149.9), and number of concomitant drugs (1.2-2.4); and (3) patient type--service (1.2-4.9), nursing division (1.5-3.8), and diagnosis related group (1.5-5.7). CONCLUSIONS: Some risk factors are consistent for all ADEs and across multiple therapeutic classes of drugs, while others are class specific. High-risk agents should be closely monitored based on patient characteristics (gender, age, weight, creatinine clearance, number of comorbidities) and drug administration (dosage, administration route, number of concomitant drugs). PMID- 15897266 TI - Interpretation of medication pictograms by adults in the UK. AB - BACKGROUND: Patients' ability to understand information about medication is crucial for safety and effectiveness. Rates of illiteracy worldwide indicate that written information alone cannot meet many patients' needs. Medication pictograms are an alternative, but may be culturally sensitive. Previous testing has used large pictograms, which are impractical for conventional drug information formats. OBJECTIVE: To compare 2 sets of pictograms for instructions or warnings (from the US and South Africa) for understandability by adults in the UK and examine the effects of pictogram size and repeat presentation on understandability among older adults. METHODS: In the first part of the study, 160 adults (aged 17-83 y) reviewed and interpreted 10 pictograms. In the second, 67 older adults (aged 65-96 y) were randomly assigned to review 10 small or large pictograms. After giving their interpretation, they were informed of the correct meaning. One week later, they were shown the same pictograms and gave their interpretation. RESULTS: The pictograms for the 10 different instructions and warnings showed great variation in interpretation rates (7.5-90%), with few significant differences between the US and South African versions. Only 3 were understood by > or = 85% of the population. Pictograms performed significantly better if they were larger and at the second presentation. CONCLUSIONS: Pictograms have the potential to help patients understand information on drug therapy. This study shows that some existing pictograms are not easily interpreted and that testing is needed before their implementation. A reduction in their size to allow incorporation into conventional written formats may cause additional problems for patients. PMID- 15897267 TI - Herbal therapy in primary health care in Maracanau, Ceara, Brazil. AB - BACKGROUND: The public health system in Brazil is insufficient to provide basic pharmaceutical and medical care to the population. Currently, some herbal therapy programs are being developed in primary healthcare services to supply the communities lacking basic medicines. OBJECTIVE: To investigate the use of herbal medicines in primary health care in Maracanau, a northeast Brazilian city. METHODS: A descriptive cross-sectional study was conducted in family health care units in Maracanau during August and September of 2002. Two hundred twenty-six patients were interviewed via structured questionnaire. Descriptive statistical analysis was accomplished using sample size and proportions. Chi2 and Student's t tests were used to compare percentages and means, respectively. RESULTS: Among the 226 patients interviewed, 144 (63.7%) reported previous use of herbal medicines. Among those, 131 (90.9%) observed therapeutic benefits from herbal medicines. We identified 10 types of herbal medicines in the prescriptions, including syrups, dyes, capsules, and ointments, for the treatment of respiratory problems (63.8%), skin conditions (15.3%), diabetes mellitus (11.4%), and other illnesses. Further, we observed that some herbal medicines were not being produced during the study period and that several patients were not aware of the fact that they were receiving herbal medicine. CONCLUSIONS: Data support the use of herbal therapy in primary health care in Maracanau. However, there is a need to conduct further study on the efficacy and safety of these herbal medicines, as well as on their quality control. PMID- 15897268 TI - Intrathymic glucocorticoid production and thymocyte survival: another piece in the puzzle. PMID- 15897269 TI - Octanoyl ghrelin is hypothalamic rooted. PMID- 15897270 TI - Cholesterol homeostasis and infertility: the liver X receptor connection. PMID- 15897272 TI - In vivo costimulatory role of B7-DC in tuning T helper cell 1 and cytotoxic T lymphocyte responses. AB - B7-DC, one of the recently described B7 family members, has the capacity to inhibit T cell responses via engagement of the immunoreceptor tyrosine-based inhibitory motif-containing inhibitory PD-1 receptor as well as enhance responses via an as yet unidentified costimulatory receptor. B7-DC is highly homologous to a coinhibitory B7 family member, B7-H1, which also binds PD-1. It is currently unclear which B7-DC function-costimulation or inhibition-predominates in vivo. To study in vivo functions of B7-DC, we evaluated immune responses in B7-DC knockout (KO) mice. Although not eliminated, interferon-gamma (IFN-gamma) production by CD4 T cells and IFN-gamma-dependent humoral responses were reduced in B7-DC KO mice relative to wild type mice. Antigen-specific CD8 T cell responses and cytotoxic T lymphocyte (CTL) activity were also diminished in B7-DC KO mice. Hepatic tumors grew more quickly in B7-DC KO mice, associated with a decrease in intrahepatic tumor-specific CD8 T cells. These results highlight the contrasting in vivo roles of B7-DC and B7-H1 and indicate that B7-DC functions as a tuning molecule, selectively augmenting T helper 1 and CTL responses. PMID- 15897273 TI - Antigen persistence is required throughout the expansion phase of a CD4(+) T cell response. AB - For CD8(+) T cells, a relatively short antigen pulse seems sufficient for antigen presenting cells to drive clonal expansion and differentiation. It is unknown whether the requirement for antigen is similarly ephemeral for CD4(+) T cells. To study the dependence of a CD4(+) T cell response on antigen persistence in a quantitatively and temporally controlled manner in vivo, we engineered a mouse line expressing a major histocompatibility complex class II-restricted epitope in dendritic cells under the control of a tetracycline-inducible promoter. Experiments tracking the proliferation of CD4(+) T cells exposed to their cognate antigen in various amounts for different time periods revealed that the division of such cells was contingent on the presence of antigen throughout their expansion phase, even in the presence of an inflammatory stimulus. This previously unrecognized feature of a CD4(+) T cell response contrasts with the proliferative behavior of CD8(+) T cells that has been documented, and it implies that the two T cell subsets might require different strategies for efficient vaccination. PMID- 15897274 TI - Shared reactivity of V{delta}2(neg) {gamma}{delta} T cells against cytomegalovirus-infected cells and tumor intestinal epithelial cells. AB - Long-lasting expansion of Vdelta2(neg) gammadelta T cells is a hallmark of cytomegalovirus (CMV) infection in kidney transplant recipients. The ligands of these cells and their role remain elusive. To better understand their immune function, we generated gammadelta T cell clones from several transplanted patients. Numerous patient Vdelta1(+), Vdelta3(+), and Vdelta5(+) gammadelta T cell clones expressing diverse Vgamma chains, but not control Vgamma9Vdelta2(+) T clones, displayed strong reactivity against CMV-infected cells, as shown by their production of tumor necrosis factor-alpha. Vdelta2(neg) gammadelta T lymphocytes could also kill CMV-infected targets and limit CMV propagation in vitro. Their anti-CMV reactivity was specific for this virus among herpesviridae and required T cell receptor engagement, but did not involve major histocompatibility complex class I molecules or NKG2D. Vdelta2(neg) gammadelta T lymphocytes expressed receptors essential for intestinal homing and were strongly activated by intestinal tumor, but not normal, epithelial cell lines. High frequencies of CMV- and tumor-specific Vdelta2(neg) gammadelta T lymphocytes were found among patients' gammadelta T cells. In conclusion, Vdelta2(neg) gammadelta T cells may play a role in protecting against CMV and tumors, probably through mucosal surveillance of cellular stress, and represent a population that is largely functionally distinct from Vgamma9Vdelta2(+) T cells. PMID- 15897275 TI - Hematopoietic cells maintain hematopoietic fates upon entering the brain. AB - Several studies have reported that bone marrow (BM) cells may give rise to neurons and astrocytes in vitro and in vivo. To further test this hypothesis, we analyzed for incorporation of neural cell types expressing donor markers in normal or injured brains of irradiated mice reconstituted with whole BM or single, purified c-kit(+)Thy1.1(lo)Lin(-)Sca-1(+) (KTLS) hematopoietic stem cells (HSCs), and of unirradiated parabionts with surgically anastomosed vasculature. Each model showed low-level parenchymal engraftment of donor-marker(+) cells with 96-100% immunoreactivity for panhematopoietic (CD45) or microglial (Iba1 or Mac1) lineage markers in all cases studied. Other than one arborizing structure in the olfactory bulb of one BM-transplanted animal, possibly representing a neuronal or glial cell process, we found no donor-marker-expressing astrocytes or non Purkinje neurons among >10,000 donor-marker(+) cells from 21 animals. These data strongly suggest that HSCs and their progeny maintain lineage fidelity in the brain and do not adopt neural cell fates with any measurable frequency. PMID- 15897276 TI - Fibrinogen and fibronectin binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis. AB - The expression of Staphylococcus aureus adhesins in Lactococcus lactis identified clumping factor A (ClfA) and fibronectin-binding protein A (FnBPA) as critical for valve colonization in rats with experimental endocarditis. This study further analyzed their role in disease evolution. Infected animals were followed for 3 d. ClfA-positive lactococci successfully colonized damaged valves, but were spontaneously eradicated over 48 h. In contrast, FnBPA-positive lactococci progressively increased bacterial titers in vegetations and spleens. At imaging, ClfA-positive lactococci were restricted to the vegetations, whereas FnBPA positive lactococci also invaded the adjacent endothelium. This reflected the capacity of FnBPA to trigger cell internalization in vitro. Because FnBPA carries both fibrinogen- and fibronectin-binding domains, we tested the role of these functionalities by deleting the fibrinogen-binding domain of FnBPA and supplementing it with the fibrinogen-binding domain of ClfA in cis or in trans. Deletion of the fibrinogen-binding domain of FnBPA did not alter fibronectin binding and cell internalization in vitro. However, it totally abrogated valve infectivity in vivo. This ability was restored in cis by inserting the fibrinogen binding domain of ClfA into truncated FnBPA, and in trans by coexpressing full length ClfA and truncated FnBPA on two separate plasmids. Thus, fibrinogen and fibronectin binding could cooperate for S. aureus valve colonization and endothelial invasion in vivo. PMID- 15897277 TI - Available carbon source influences the resistance of Neisseria meningitidis against complement. AB - Neisseria meningitidis is an important cause of septicaemia and meningitis. To cause disease, the bacterium must acquire essential nutrients for replication in the systemic circulation, while avoiding exclusion by host innate immunity. Here we show that the utilization of carbon sources by N. meningitidis determines its ability to withstand complement-mediated lysis, through the intimate relationship between metabolism and virulence in the bacterium. The gene encoding the lactate permease, lctP, was identified and disrupted. The lctP mutant had a reduced growth rate in cerebrospinal fluid compared with the wild type, and was attenuated during bloodstream infection through loss of resistance against complement-mediated killing. The link between lactate and complement was demonstrated by the restoration of virulence of the lctP mutant in complement (C3(-/-))-deficient animals. The underlying mechanism for attenuation is mediated through the sialic acid biosynthesis pathway, which is directly connected to central carbon metabolism. The findings highlight the intimate relationship between bacterial physiology and resistance to innate immune killing in the meningococcus. PMID- 15897278 TI - TGF-{beta}-dependent CD103 expression by CD8(+) T cells promotes selective destruction of the host intestinal epithelium during graft-versus-host disease. AB - Destruction of the host intestinal epithelium by donor effector T cell populations is a hallmark of graft-versus-host disease (GVHD), but the underlying mechanisms remain obscure. We demonstrate that CD8(+) T cells expressing CD103, an integrin conferring specificity for the epithelial ligand E-cadherin, play a critical role in this process. A TCR transgenic GVHD model was used to demonstrate that CD103 is selectively expressed by host-specific CD8(+) T cell effector populations (CD8 effectors) that accumulate in the host intestinal epithelium during GVHD. Although host-specific CD8 effectors infiltrated a wide range of host compartments, only those infiltrating the intestinal epithelium expressed CD103. Host-specific CD8 effectors expressing a TGF-beta dominant negative type II receptor were defective in CD103 expression on entry into the intestinal epithelium, which indicates local TGF-beta activity as a critical regulating factor. Host-specific CD8 effectors deficient in CD103 expression successfully migrated into the host intestinal epithelium but were retained at this site much less efficiently than wild-type host-specific CD8 effectors. The relevance of these events to GVHD pathogenesis is supported by the finding that CD103-deficient CD8(+) T cells were strikingly defective in transferring intestinal GVHD pathology and mortality. Collectively, these data document a pivotal role for TGF-beta-dependent CD103 expression in dictating the gut tropism, and hence the destructive potential, of CD8(+) T cells during GVHD pathogenesis. PMID- 15897279 TI - Impaired early B cell tolerance in patients with rheumatoid arthritis. AB - Autoantibody production is a characteristic of most autoimmune diseases including rheumatoid arthritis (RA). The role of these autoantibodies in the pathogenesis of RA remains elusive, but they appear in the serum many years before the onset of clinical disease suggesting an early break in B cell tolerance. The stage of B cell development at which B cell tolerance is broken in RA remains unknown. We previously established in healthy donors that most polyreactive developing B cells are silenced in the bone marrow, and additional autoreactive B cells are removed in the periphery. B cell tolerance in untreated active RA patients was analyzed by testing the specificity of recombinant antibodies cloned from single B cells. We find that autoreactive B cells fail to be removed in all six RA patients and represent 35-52% of the mature naive B cell compartment compared with 20% in healthy donors. In some patients, RA B cells express an increased proportion of polyreactive antibodies that can recognize immunoglobulins and cyclic citrullinated peptides, suggesting early defects in central B cell tolerance. Thus, RA patients exhibit defective B cell tolerance checkpoints that may favor the development of autoimmunity. PMID- 15897280 TI - Induction of vascular leakage through release of bradykinin and a novel kinin by cysteine proteinases from Staphylococcus aureus. AB - Staphylococcus aureus is a major pathogen of gram-positive septic shock and frequently is associated with consumption of plasma kininogen. We examined the vascular leakage (VL) activity of two cysteine proteinases that are secreted by S. aureus. Proteolytically active staphopain A (ScpA) induced VL in a bradykinin (BK) B(2)-receptor-dependent manner in guinea pig skin. This effect was augmented by staphopain B (SspB), which, by itself, had no VL activity. ScpA also produced VL activity from human plasma, apparently by acting directly on kininogens to release BK, which again was augmented significantly by SspB. Intravenous injection of ScpA into a guinea pig caused BK B(2)-receptor-dependent hypotension. ScpA and SspB together induced the release of leucyl-methionyl-lysyl BK, a novel kinin with VL and blood pressure-lowering activities that are equivalent to BK. Collectively, these data suggest that production of BK and leucyl-methionyl-lysyl-BK by staphopains is a new mechanism of S. aureus virulence and bacterial shock. Therefore, staphopain-specific inhibitors and kinin-receptor antagonists could be used to treat this disease. PMID- 15897281 TI - I{kappa}B kinase (IKK){beta}, but not IKK{alpha}, is a critical mediator of osteoclast survival and is required for inflammation-induced bone loss. AB - Transcription factor, nuclear factor kappaB (NF-kappaB), is required for osteoclast formation in vivo and mice lacking both of the NF-kappaB p50 and p52 proteins are osteopetrotic. Here we address the relative roles of the two catalytic subunits of the IkappaB kinase (IKK) complex that mediate NF-kappaB activation, IKKalpha and IKKbeta, in osteoclast formation and inflammation induced bone loss. Our findings point out the importance of the IKKbeta subunit as a transducer of signals from receptor activator of NF-kappaB (RANK) to NF kappaB. Although IKKalpha is required for RANK ligand-induced osteoclast formation in vitro, it is not needed in vivo. However, IKKbeta is required for osteoclastogenesis in vitro and in vivo. IKKbeta also protects osteoclasts and their progenitors from tumor necrosis factor alpha-induced apoptosis, and its loss in hematopoietic cells prevents inflammation-induced bone loss. PMID- 15897282 TI - Linear and logarithmic subtraction for detecting enamel subsurface demineralization. AB - OBJECTIVE: To compare the diagnostic performance of linear and logarithmically contrast-enhanced subtraction images, acquired with digital and digitized radiographs, in detecting approximal enamel subsurface demineralization. METHODS: Fifty caries-free human third molars were immersed in a demineralizing solution for 60, 75, 90, and 120 days, in order to induce artificial enamel subsurface demineralization. The teeth were coated with nail varnish, leaving only a circular window of approximately 7 mm2 in one of the approximal surfaces, allowing contact with the solution. Standardized radiographs of the teeth were taken prior to and after the demineralization period with three digital systems, CygnusRay MPS, DenOptix and DIGORA, and InSight film. Conventional, digital and digitized radiographs were assessed by three experienced radiologists. Linear and logarithmically contrast-enhanced subtraction images were acquired and then examined by a fourth independent radiologist. For the validation of the radiographic diagnosis, the enamel test areas were submitted to Knoop microhardness profiling. Radiographic interpretation data was evaluated using ROC analysis. The areas under the ROC curves (Az) were compared by the chi-squared test. The level of significance was set at P=0.05. RESULTS: No significant differences were found between linear and logarithmically contrast- enhanced subtraction images, acquired with the four studied modalities: CygnusRay MPSlinear (Az=0.95), CygnusRay MPSlog (Az=0.98), DenOptixlinear (Az=0.97), DenOptixlog (Az=0.99), DIGORAlinear (Az=0.98), DIGORAlog (Az=0.98), digitized radiographylinear (Az=0.99), digitized radiographylog (Az=0.99). CONCLUSION: Linear and logarithmically contrast-enhanced subtraction images, acquired with digital and digitized radiographs, were diagnostically comparable for assessing enamel subsurface demineralization. PMID- 15897283 TI - Effects of the automatic exposure compensation on the proximal caries diagnosis. AB - OBJECTIVE: To elucidate the effect of automatic exposure compensation (AEC) on the diagnostic accuracy of proximal caries by comparing several digital intraoral imaging systems with a film. MATERIALS AND METHODS: Twenty-seven extracted teeth served as proximal caries samples. Three digital radiographic systems; the Compuray, the Dixel, and the Sens-A-Ray without scintillator layer, and Kodak Ekta-speed Plus films were used as recording media. Radiographs of the teeth samples were obtained with each recording medium under seven to eight different exposures including the optimum level. Six oral radiologists evaluated the possibility of proximal caries with the five-grade-confidence-scale. On digital radiographs, image manipulations were allowed after the initial assessment. Receiver operating characteristic (ROC) curves were obtained at each exposure in each recording medium. The area under the ROC curve (Az) was used as the representative value of diagnostic accuracy. Diagnostic accuracy (DA) curves were obtained by plotting averaged Az values from all observers as a function of incident exposure in each system. RESULTS: The effect of exposure variation on the DA was slight in the film while it was significantly larger in the digital systems without AEC. Among digital systems, the effect of exposure variation was smaller in the system with AEC than those without AEC. There was no significant effect on the diagnostic accuracy even if digital image manipulation was employed. CONCLUSION: AEC minimizes the decrease of DA due to inadequate exposures. Since it compensates for the narrow exposure range in the digital intraoral sensor systems, the system with AEC may be preferable for the clinical diagnostic tasks. PMID- 15897284 TI - Dosimetry of digital panoramic imaging. Part I: Patient exposure. AB - OBJECTIVES: To measure patient radiation dose during panoramic exposure with various panoramic units for digital panoramic imaging. METHODS: An anthropomorphic phantom was filled with thermoluminescent dosemeters (TLD 100) and exposed with five different digital panoramic units during ten consecutive exposures. Four machines were equipped with a direct digital CCD (charge coupled device) system, whereas one of the units used storage phosphor plates (indirect digital technique). The exposure settings recommended by the different manufacturers for the particular image and patient size were used: tube potential settings ranged between 64 kV and 74 kV, exposure times between 8.2 s and 19.0 s, at fuse current values between 4 mA and 7 mA. The effective radiation dose was calculated with inclusion of the salivary glands. RESULTS: Effective radiation doses ranged between 4.7 microSv and 14.9 microSv for one exposure. Salivary glands absorbed the most radiation for all panoramic units. When indirect and direct digital panoramic systems were compared, the effective dose of the indirect digital unit (8.1 microSv) could be found within the range of the effective doses for the direct digital units (4.7-14.9 microSv). CONCLUSIONS: A rather wide range of patient radiation doses can be found for digital panoramic units. There is a tendency for lower effective doses for digital compared with analogue panoramic units, reported in previous studies. PMID- 15897285 TI - Dosimetry of digital panoramic imaging. Part II: Occupational exposure. AB - OBJECTIVES: To measure occupational radiation dose during panoramic exposure from five digital panoramic X-ray units. METHODS: Exposures were made with five different digital panoramic units, of which four were equipped with a direct digital CCD (charge coupled device, "direct digital" technique), and one used storage phosphor plates ("indirect digital" technique). An anthropomorphic phantom served as the patient. An ionization chamber recorded the scattered radiation at 1 m from the phantom at five different locations around the panoramic units, both at the level of the thyroid gland and the level of the gonads, and effective organ doses were calculated. Exposure parameters were set as recommended by the manufacturers for the particular image and patient size: tube potential settings ranged between 64 kV and 74 kV, exposure times between 8.2 s and 19.0 s, tube current values between 4 mA and 7 mA. RESULTS: The maximum organ equivalent dose at 1 m from the panoramic unit was 0.60 microGy, the maximum organ effective dose was 0.10 microSv. Organ equivalent doses varied between 0.18 microGy and 0.30 microGy and organ effective doses between 0.01 microSv and 0.05 microSv for the different positions around the units (average for the different panoramic units). The variations in organ doses for the various machines were 0.04-0.53 microGy organ equivalent dose and 0.01-0.08 microSv organ effective dose. CONCLUSIONS: Assuming that 500 panoramic radiographs per year are taken by a dental practitioner at 1 m distance from the panoramic unit, he or she will receive an annual additional organ effective dose between 5 microSv and 15 microSv for the thyroid gland and between 5 microSv and 40 microSv for the gonads, depending on the type of digital panoramic unit. PMID- 15897286 TI - Narrowing of the inferior dental canal in relation to the lower third molars. AB - OBJECTIVES: To assess narrowing of the inferior dental canal in the lower third molar regions using computed tomography (CT) and to determine the value of radiographic markers on rotational panoramic radiographs in assessing the true relationships of the inferior dental canal. METHODS: Patients referred for CT assessment of impacted lower third molars were used in this study. The lower third molars were assessed using CT to determine the position and morphology of the inferior dental canal relative to the roots and the cortical plates. The radiographic markers on rotational panoramic radiographs were correlated with the CT findings when rotational panoramic radiographs were available. RESULTS: The patients referred had 202 lower third molars. Inferior positioning of the inferior dental canal was the most common location on CT. Narrowing of the inferior dental canal was found in relation to the lower third molars in 66.8% of cases. The chance of narrowing of the inferior dental canal as shown using CT increased when at least one of the radiographic markers, superimposition, narrowing, deviation or reduction in density was present on the rotational panoramic radiograph. Deviation of the inferior dental canal on rotational panoramic radiographs was found to be the most significant predictor of narrowing of the inferior dental canal and a close relationship to the roots, as shown in CT. CONCLUSIONS: Narrowing of the inferior dental canal is a common finding when impacted lower third molars are assessed using CT. On rotational panoramic radiographs deviation of the inferior dental canal is the best predictor of narrowing of the inferior dental canal and a close relationship to the roots. PMID- 15897287 TI - Spiral computed tomographic demonstration of C-shaped canals in mandibular second molars. AB - OBJECTIVES: The aim of this study was to evaluate the prevalence of C-shaped canals in single rooted mandibular second molars with spiral computed tomographic imaging in detail. METHODS: One hundred and twelve single-rooted mandibular second molar samples were selected from 491 extracted mandibular second molars. The teeth were scanned by spiral computed tomographic technique to examine canal configurations. RESULTS: The evaluation criteria of the shape of roots was based on Manning's category as round, oval and C-shaped. The prevalence of C-shaped canals was found to be 8.1% in mandibular second molars. The evaluation of the root canal configurations of C-shaped mandibular second molars with single roots (40 teeth) was based on Vertucci's classification. Vertucci type I canals were most frequently seen in C-shaped mandibular second molars (40%). Additionally, we disclosed three additional canal types. CONCLUSION: Results of this study demonstrated that high resolution visualization of the root canal shape can be achieved by spiral computed tomographic imaging. PMID- 15897288 TI - Change in mandibular trabecular pattern and hip fracture rate in elderly women. AB - OBJECTIVES: To determine if alterations of trabecular pattern, or the rate of change of jaw trabeculae, are associated with rate of hip fracture. METHODS: Participants in a population-based study of residents of a California retirement community (Leisure World Cohort Study) were asked for permission to obtain their dental radiographs. Periapical radiographs were retrieved on 598 women (average age at time of first radiograph=77 years). Several measurements of trabecular pattern (strut analysis), textural properties (run-length analysis) and Fourier analysis were made in several anatomical regions of the jaw. These trabecular features and clinical information self-reported by subjects in the early 1980s were examined for association with hip fracture rate using Cox proportional hazard regression. RESULTS: Rate of hip fracture increased with decreasing average length of node-to-terminus struts in the mandibular incisor region. Each 0.01 mm per year decrease in the average length of node-to-terminus struts increased hip fracture rate by a factor of 2.9 (P=0.02, accuracy=73%). Inclusion of clinical parameters improved the predictive model compared with use of the radiographic parameter alone (accuracy=79%). Similar results were seen for percent change per year in this parameter. CONCLUSIONS: Changes in radiographic trabecular structure, augmented with clinical information, are predictive of hip fracture in elderly women. Further refinement of both the radiographic and clinical parameters may lead to a screening process accessible to a large number of women and to early diagnosis and treatment of osteoporosis. PMID- 15897289 TI - Temporomandibular joint internal derangement: relationship between joint pain and MR grading of effusion and total protein concentration in the joint fluid. AB - OBJECTIVES: The objective of this study was to determine whether there is any association between the protein concentration in the synovial fluid and (i) the amount of articular hydrops, as graded in magnetic resonance (MR) images, and (ii) joint pain in temporomandibular joints (TMJs) with and without displacement of the disc. PATIENTS AND METHODS: This study involved 16 joints in 16 patients referred to our clinic with the complaint of pain and limited mouth opening. The control group consisted of 15 joints in 15 patients with unilaterally normal disc and condyle relationship and no pain while the opposite side had a non-painful joint with disc displacement without reduction (DDwoR). The subjects and controls were different individuals and only a single joint was used for each. Pain and dysfunction were evaluated by visual analogue scale. Bilateral proton density and T2 weighted images of the TMJs of the 31 subjects were analysed for fluid and condyle bony changes as well as disc position. The amount of fluid, identified as an area of high signal intensity in the region of the upper and lower joint spaces, was characterized as none, minimal, moderate or marked. Arthrocentesis was performed both for synovial fluid analysis of total protein concentration and the treatment of the joints with DDwoR. Total protein concentration was measured by using protein dye binding on spectrophotometry. RESULTS: All patients experienced a significant (P<0.01) increase in maximal mouth opening immediately post-arthrocentesis. In the study group, the disc was displaced most frequently in an anteriormedial direction (75%) and deformation of disc form was seen in 13 joints. Condylar bony changes were seen in 27% of joints in the control group and in 81% of joints in the study group. A statistically significant association was found between joints with disc displacement, disc form and condylar bony changes (P<0.05). In the control group, only one joint, which had an osteophyte, showed joint effusion (JE) with moderate fluid. In the study group, only four joints had no fluid (25%). JE was found in 10 (63%) joints with disc displacement on anteromedial direction, in 10 (63%) joints with disc deformation and in 10 (63%) joints with osteophytes and erosion. Mean total protein concentration was 16.87+/ 7.9 (range 7.4-34.1 mg dl-1) in control joints, 55.08+/-35.16 (range 21.5-153.9 mg dl-1) in study joints. There were significant differences in the mean total protein concentration between the control and study groups (P<0.01). Significant positive correlation was found between the total protein concentration and JE (r=0.65, P<0.01). No significant correlation was found between the level of pain and dysfunction and JE and total protein concentration in either control or study groups (P>0.05). CONCLUSION: Pain in the TMJ was not related to MR findings of effusion in internal derangement and synovial fluid aspirate findings of total protein concentration. However, total protein concentration was related to the amount of JE in DDwoR joints and painful joints were more likely to demonstrate the JE. PMID- 15897290 TI - The clinical and diagnostic imaging findings of osteosarcoma of the jaw. AB - OBJECTIVE: To clarify the valuable clinical features and diagnostic imaging findings regarding the diagnosis of osteosarcoma of the jaw (OSJ). MATERIALS AND METHODS: The initial symptoms and diagnostic imaging findings of 10 patients with OSJ were analysed. The points analysed on the diagnostic images were as follows: any widening of the periodontal ligament space of the teeth on the periphery of the OSJ; the presence of radial spicules and Codman's triangle; any signs of bone destruction; and the patterns of osteogenesis. RESULTS: All patients had pain and/or swelling of the affected site, and all OSJs, except for one edentulous case, showed a widening of the periodontal ligament space of the teeth on the periphery of the OSJ. Radial spicules or Codman's triangle were observed in only three cases (30%). Four out of five mandibular OSJs were osteolytic or osteolytic dominant with bone destruction, while, in contrast, four out of five maxillary OSJs were osteogenic or osteogenic dominant, and three out of the four maxillary OSJs did not show bone destruction. The osteogenic OSJ without bone destruction was similar to some benign cemento-osseous lesions of the jaw and thus was difficult to diagnose as OSJ based on the diagnostic imaging findings alone. CONCLUSION: Even though some OSJ showed features similar to the benign tumours of the jaw bone based on the diagnostic imaging findings, the pain and swelling of the affected site, and the widening of the periodontal ligament space of the teeth on the periphery of OSJ were considered to be common findings, which may help in making an accurate diagnosis of OSJ in this limited series. PMID- 15897291 TI - Emergency maxillofacial radiology. Foreign body localization: report of cases. AB - Successful emergency removal of foreign bodies of the maxillofacial complex depends on the surgeon's ability to perform exact location procedures. In hospitals and/or offices where advanced imaging technologies are not available, the attendant clinician has to use and rely on variations of conventional radiographic techniques in order to complement the diagnosis and locate the foreign body. Three cases are presented where unconventional radiographic techniques were used, leading to successful surgical removal of foreign bodies. PMID- 15897292 TI - Osteochondritis dissecans of the temporomandibular joint. AB - A case is reported of a 43-year-old female patient presenting bilateral osteochondritis dissecans (OCD) of the temporomandibular joint (TMJ), in different stages for each side, associated with avascular necrosis (AVN) of the right condyle. Additionally observed was anterior disk displacement without reduction for both sides. We have proposed an adaptation of the previous classification of OCD for cases affecting the TMJ. We have also stressed the fundamental role of panoramic radiography on the diagnosis of stage 3 and stage 4 OCD of the TMJ. In relation to MRI, we have recommended sagittal (slice thickness of 2 mm) and coronal (slice thickness of 1 mm) fast spin-echo proton density weighted sequences to better identify bone lesions (stage 1 and 2) and also localize osteochondral loose bodies; and coronal (slice thickness of 1 mm) fat suppressed fast spin-echo T2 weighted sequence to better evaluate OCD (stable or unstable) and the features of the occasionally associated AVN (acute or chronic). PMID- 15897293 TI - Two stable, conducting conformations of the selectivity filter in Shaker K+ channels. AB - We have examined the voltage dependence of external TEA block of Shaker K(+) channels over a range of internal K(+) concentrations from 2 to 135 mM. We found that the concentration dependence of external TEA block in low internal K(+) solutions could not be described by a single TEA binding affinity. The deviation from a single TEA binding isotherm was increased at more depolarized membrane voltages. The data were well described by a two-component binding scheme representing two, relatively stable populations of conducting channels that differ in their affinity for external TEA. The relative proportion of these two populations was not much affected by membrane voltage but did depend on the internal K(+) concentration. Low internal K(+) promoted an increase in the fraction of channels with a low TEA affinity. The voltage dependence of the apparent high-affinity TEA binding constant depended on the internal K(+) concentration, becoming almost voltage independent in 5 mM. The K(+) sensitivity of these low- and high-affinity TEA states suggests that they may represent one- and two-ion occupancy states of the selectivity filter, consistent with recent crystallographic results from the bacterial KcsA K(+) channel. We therefore analyzed these data in terms of such a model and found a large (almost 14-fold) difference between the intrinsic TEA affinity of the one-ion and two-ion modes. According to this analysis, the single ion in the one-ion mode (at 0 mV) prefers the inner end of the selectivity filter twofold more than the outer end. This distribution does not change with internal K(+). The two ions in the two-ion mode prefer to occupy the inner end of the selectivity filter at low K(+), but high internal K(+) promotes increased occupancy of the outer sites. Our analysis further suggests that the four K(+) sites in the selectivity filter are spaced between 20 and 25% of the membrane electric field. PMID- 15897294 TI - The amino terminus of Slob, Slowpoke channel binding protein, critically influences its modulation of the channel. AB - The Drosophila Slowpoke calcium-dependent potassium channel (dSlo) binding protein Slob was discovered by a yeast two-hybrid screen using the carboxy terminal tail region of dSlo as bait. Slob binds to and modulates the dSlo channel. We have found that there are several Slob proteins, resulting from multiple translational start sites and alternative splicing, and have named them based on their molecular weights (in kD). The larger variants, which are initiated at the first translational start site and are called Slob71 and Slob65, shift the voltage dependence of dSlo activation, measured by the whole cell conductance-voltage relationship, to the left (less depolarized voltages). Slob53 and Slob47, initiated at the third translational start site, also shift the dSlo voltage dependence to the left. In contrast, Slob57 and Slob51, initiated at the second translational start site, shift the conductance-voltage relationship of dSlo substantially to more depolarized voltages, cause an apparent dSlo channel inactivation, and increase the deactivation rate of the channel. These results indicate that the amino-terminal region of Slob plays a critical role in its modulation of dSlo. PMID- 15897295 TI - Cysteine accessibility in ClC-0 supports conservation of the ClC intracellular vestibule. AB - ClC chloride channels, which are ubiquitously expressed in mammals, have a unique double-barreled structure, in which each monomer forms its own pore. Identification of pore-lining elements is important for understanding the conduction properties and unusual gating mechanisms of these channels. Structures of prokaryotic ClC transporters do not show an open pore, and so may not accurately represent the open state of the eukaryotic ClC channels. In this study we used cysteine-scanning mutagenesis and modification (SCAM) to screen >50 residues in the intracellular vestibule of ClC-0. We identified 14 positions sensitive to the negatively charged thiol-modifying reagents sodium (2 sulfonatoethyl)methanethiosulfonate (MTSES) or sodium 4-acetamido-4' maleimidylstilbene-2'2-disulfonic acid (AMS) and show that 11 of these alter pore properties when modified. In addition, two MTSES-sensitive residues, on different helices and in close proximity in the prokaryotic structures, can form a disulfide bond in ClC-0. When mapped onto prokaryotic structures, MTSES/AMS sensitive residues cluster around bound chloride ions, and the correlation is even stronger in the ClC-0 homology model developed by Corry et al. (2004). These results support the hypothesis that both secondary and tertiary structures in the intracellular vestibule are conserved among ClC family members, even in regions of very low sequence similarity. PMID- 15897296 TI - Microbial K+ channels. PMID- 15897298 TI - Adiponectin and adiponectin receptors. AB - Metabolic syndrome is thought to result from obesity and obesity-linked insulin resistance. Obesity in adulthood is characterized by adipocyte hypertrophy. Adipose tissue participates in the regulation of energy homeostasis as an important endocrine organ that secretes a number of biologically active "adipokines."Heterozygous peroxisome proliferator-activated receptor-gamma knockout mice were protected from high-fat diet induced obesity, adipocyte hypertrophy, and insulin resistance. Systematic gene profiling analysis of these mice revealed that adiponectin/Acrp30 was overexpressed. Functional analyses including generation of adiponectin transgenic or knockout mice have revealed that adiponectin serves as an insulin-sensitizing adipokine. In fact, obesity linked down-regulation of adiponectin was a mechanism whereby obesity could cause insulin resistance and diabetes. Recently, we have cloned adiponectin receptors in the skeletal muscle (AdipoR1) and liver (AdipoR2), which appear to comprise a novel cell-surface receptor family. We showed that AdipoR1 and AdipoR2 serve as receptors for globular and full-length adiponectin and mediate increased AMP activated protein kinase, peroxisome proliferator-activated receptor-alpha ligand activities, and glucose uptake and fatty-acid oxidation by adiponectin. Obesity decreased expression levels of AdipoR1/R2, thereby reducing adiponectin sensitivity, which finally leads to insulin resistance, the so-called "vicious cycle." Most recently, we showed that osmotin, which is a ligand for the yeast homolog of AdipoR (PHO36), activated AMPK via AdipoR in C2C12 myocytes. This may facilitate efficient development of adiponectin receptor agonists. Adiponectin receptor agonists and adiponectin sensitizers should serve as versatile treatment strategies for obesity-linked diseases such as diabetes and metabolic syndrome. PMID- 15897299 TI - In the middle of the decade. PMID- 15897300 TI - The needle and the damage done. PMID- 15897301 TI - Benefit of pregnancy in inflammatory arthritis. PMID- 15897302 TI - Pulmonary arterial hypertension in systemic sclerosis: diagnostic pathway and therapeutic approach. PMID- 15897303 TI - Proteolysis of serum amyloid A and AA amyloid proteins by cysteine proteases: cathepsin B generates AA amyloid proteins and cathepsin L may prevent their formation. AB - BACKGROUND: AA amyloidosis develops in patients with chronic inflammatory diseases. The AA amyloid proteins are proteolytic fragments obtained from serum amyloid A (SAA). Previous studies have provided evidence that endosomes or lysosomes might be involved in the processing of SAA, and contribute to the pathology of AA amyloidosis. OBJECTIVE: To investigate the anatomical distribution of cathepsin (Cath) B and CathL in AA amyloidosis and their ability to process SAA and AA amyloid proteins. METHODS: and results: CathB and CathL were found immunohistochemically in every patient with AA amyloidosis and displayed a spatial relationship with amyloid in all the cases studied. Both degraded SAA and AA amyloid proteins in vitro. With the help of mass spectrometry 27 fragments were identified after incubation of SAA with CathB, nine of which resembled AA amyloid proteins, and seven fragments after incubation with CathL. CathL did not generate AA amyloid-like peptides. When native human AA amyloid proteins were used as a substrate 26 fragments were identified after incubation with CathB and 18 after incubation with CathL. CONCLUSION: The two most abundant and ubiquitously expressed lysosomal proteases can cleave SAA and AA amyloid proteins. CathB generates nine AA amyloid-like proteins by its carboxypeptidase activity, whereas CathL may prevent the formation of AA amyloid proteins by endoproteolytic activity within the N-terminal region of SAA. This is particularly interesting, because AA amyloidosis is a systemic disease affecting many organs and tissue types, almost all of which express CathB and CathL. PMID- 15897304 TI - Doppler ultrasound findings in healthy wrists and finger joints before and after use of two different contrast agents. AB - OBJECTIVE: To examine the effect of contrast agents on Doppler ultrasound findings in the synovial membrane in the wrist and fingers of healthy volunteers. MATERIALS AND METHODS: Eleven healthy subjects were included in the study (5 women and 6 men, mean age 38 years, range (20-60)). They had no clinical signs of inflammatory or degenerative joint diseases. A total of 66 joints were examined-6 joints for each subject: wrist and metacarpophalangeal (MCP) joints 1-5-before contrast injection and after Levovist and SonoVue injection with a 30 minute interval. RESULTS: Colour Doppler activity was detected in 10/55 (18%) MCP joints before contrast injection and in 29/55 (53%) and 28/55 (51%) joints after Levovist (p<0.0001) and SonoVue injection (p = 0.0001), respectively. A significant increase in Doppler activity in the radial (p<0.05) and ulnar (p = 0.01) parts of the wrist joint was detected only after SonoVue injections. With spectral Doppler no difference was found in the resistive index (RI) in the vessels measured before as compared with those only detected after contrast injection. CONCLUSION: The number of joints with colour Doppler activity in healthy volunteers was increased by the use of contrast agents. No changes in RI were detected. The value of contrast agents remains to be demonstrated in inflammatory diagnostics. PMID- 15897305 TI - Immunosuppressive treatment of chronic periaortitis: a retrospective study of 20 patients with chronic periaortitis and a review of the literature. AB - BACKGROUND: Retroperitoneal fibrosis (RPF) and inflammatory aneurysm of the abdominal aorta (IAAA) are regarded as two manifestations of the same disease, termed "chronic periaortitis". OBJECTIVE: To determine the optimal therapeutic and diagnostic approaches to IAAA. METHODS: The outcome of medical immunosuppressive and surgical treatment of 20 patients was examined. Measurements of the C reactive protein (CRP) were compared with contrast enhanced imaging studies in the follow up of the patients. RESULTS: The diameter of the periaortic mantle and its contrast enhancement improved in 13/15 (87%) patients given immunosuppressive treatment for a period of more than 6 months. Strong contrast enhancement was associated with a substantial rise in CRP, but no correlation between the CRP value and thickness of the fibrotic mass was found, even at intraindividual follow up. CONCLUSIONS: Immunosuppressive treatment should be included in the first line treatment of patients with RPF and should be maintained long term. Imaging studies are better than CRP measurements in the evaluation of response to treatment. PMID- 15897306 TI - Raised levels of interleukin 6 in systemic lupus erythematosus correlate with anaemia. AB - OBJECTIVES: Raised levels of the cytokines interleukin (IL) 6 and IL10 have been reported in patients with systemic lupus erythematosus (SLE). OBJECTIVE: To determine if levels of IL6 and IL10 correlate with organ/system-specific disease activity in SLE, using the British Isles Lupus Assessment Group (BILAG) Disease Activity Index. METHODS: Levels of IL6 and IL10 in serum samples from 171 patients with SLE and 50 normal controls were determined by enzyme linked immunosorbent assay (ELISA). Levels of cytokines in individual patients with SLE were compared with the presence or absence of active disease in eight organ/systems using the BILAG index. RESULTS: Levels of IL6 were significantly higher (p = 0.005) in patients with active compared with inactive haematological disease, as scored by the BILAG index. Further analysis showed that this association was dependent on an inverse correlation (p = 0.002, r = -0.26) between IL6 levels and haemoglobin levels in patients with SLE. In contrast, IL10 levels did not correlate with individual organ/system disease activity. CONCLUSIONS: Raised levels of IL6 in SLE may influence the development of anaemia in this disease. These findings are in agreement with an increasing number of studies, which support physiological links between IL6 and anaemia. Importantly, with the exception of the haematological system, our studies do not provide evidence of any individual organ/system which would respond to therapeutic manipulation of either IL6 or IL10 levels. PMID- 15897307 TI - Uveitis in young adults with juvenile idiopathic arthritis: a clinical evaluation of 123 patients. AB - OBJECTIVE: To examine the prevalence and characteristics of uveitis in young adults with juvenile idiopathic arthritis (JIA). METHODS: The study population consisted of 123 JIA patients born between 1976 and 1980 whose arthritis had been diagnosed and treatment first started at the Rheumatism Foundation Hospital in 1976 to 1995. A clinical re-evaluation was carried out by an ophthalmologist and a paediatric rheumatologist 16 years later on average. RESULTS: The mean age of the patients was 23.5 years, 72% were women, and 63% had oligoarthritis. During the course of the disease, diagnosis of uveitis had been made in 25 patients (20%). Arthritis in the 19 patients with asymptomatic uveitis was more often ongoing than in the 98 patients without uveitis (p = 0.032). Asymptomatic uveitis was persistent in eight of the 19 cases (42%), and arthritis was active in seven of these. Four of the six patients with attacks of symptomatic uveitis had parallel treatment for arthritis. In three of 19 patients with asymptomatic uveitis and in five of six with acute uveitis the eye inflammation had started after the age of 16. At the onset of arthritis the patients with asymptomatic uveitis were younger than those without uveitis (p = 0.002). Complications of uveitis developed in six patients but their sight remained good. CONCLUSIONS: Asymptomatic uveitis continued into adulthood in almost half the uveitis patients. Most also had ongoing arthritis. Acute uveitis was often associated with persistent arthritis. PMID- 15897308 TI - Favourable social functioning and health related quality of life of patients with JIA in early adulthood. AB - OBJECTIVE: To evaluate the social functioning and health related quality of life (HRQoL) in patients with juvenile idiopathic arthritis (JIA) in early adulthood. METHODS: The patient files of the Rheumatism Foundation Hospital were screened to identify patients born in 1976-1980 diagnosed as having JIA. HRQoL was measured by the RAND 36-item health survey 1.0; spousal relationships and educational and employment status were assessed by questionnaire. The patients were invited to a follow up study. Age and sex matched controls from the community were identified in the Finnish population registry. RESULTS: Of 187 patients identified, 123 participated. Spousal relationships, educational level, and employment status were similar to controls. HRQoL in JIA patients was similar to controls except on the physical functioning scale. At follow up 35% of patients were in remission. Patients with active disease had poorer HRQoL in the physical component than those in remission or controls. The extended oligoarthritis group had the lowest physical and mental score in HRQoL compared with the other JIA subgroups. The patient's own evaluation was the explanatory factor in both the physical and mental component of HRQoL. CONCLUSION: Social functioning and HRQoL were similar in JIA patients and age, sex, and municipality matched controls. However, patients with extended oligoarthritis attained significantly lower scores in the physical and mental component of HRQoL than oligo- or polyarthritis patients. Special attention in everyday care should be paid to those patients who have active disease or the extended oligoarthritis type of disease. PMID- 15897309 TI - Increased pentosidine, an advanced glycation end product, in serum and synovial fluid from patients with knee osteoarthritis and its relation with cartilage oligomeric matrix protein. AB - BACKGROUND: Pentosidine, an advanced glycation end product, increasingly accumulates in articular cartilage with age, and contributes to the pathogenesis of osteoarthritis (OA). Increased pentosidine concentrations are associated with inflammatory disorders-for example, rheumatoid arthritis. OBJECTIVE: To compare pentosidine serum concentrations in patients with knee OA and in healthy volunteers and to determine a relationship between pentosidine and cartilage oligomeric matrix protein (COMP)-a marker of articular cartilage destruction. METHODS: Paired serum and synovial fluid samples were obtained by arthrocentesis from 38 patients with knee OA and from 38 healthy volunteers. Pentosidine concentration was measured by reverse phase high performance liquid chromatography with fluorescent detection and COMP was determined by sandwich ELISA. RESULTS: Significantly increased serum pentosidine (p<0.01) and COMP (p<0.05) levels were detected in the patients with OA compared with the control group. Serum pentosidine correlated significantly with synovial fluid pentosidine (p<0.001). Pentosidine in synovial fluid (p<0.05) and in serum (p<0.05) correlated significantly with synovial fluid COMP. Pentosidine and COMP concentrations did not correlate significantly with the radiological stage of the disease. CONCLUSION: Increased pentosidine serum concentration in patients with OA and its correlation with the cartilage destruction marker COMP in synovial fluid suggests that pentosidine may be important in OA pathology and is a new potential OA marker. PMID- 15897310 TI - Efficacy of physiotherapy management of knee joint osteoarthritis: a randomised, double blind, placebo controlled trial. AB - OBJECTIVE: To determine whether a multimodal physiotherapy programme including taping, exercises, and massage is effective for knee osteoarthritis, and if benefits can be maintained with self management. METHODS: Randomised, double blind, placebo controlled trial; 140 community volunteers with knee osteoarthritis participated and 119 completed the trial. Physiotherapy and placebo interventions were applied by 10 physiotherapists in private practices for 12 weeks. Physiotherapy included exercise, massage, taping, and mobilisation, followed by 12 weeks of self management. Placebo was sham ultrasound and light application of a non-therapeutic gel, followed by no treatment. Primary outcomes were pain measured by visual analogue scale and patient global change. Secondary measures included WOMAC, knee pain scale, SF-36, assessment of quality of life index, quadriceps strength, and balance test. RESULTS: Using an intention to treat analysis, physiotherapy and placebo groups showed similar pain reductions at 12 weeks: -2.2 cm (95% CI, -2.6 to -1.7) and -2.0 cm (-2.5 to -1.5), respectively. At 24 weeks, pain remained reduced from baseline in both groups: 2.1 (-2.6 to -1.6) and -1.6 (-2.2 to -1.0), respectively. Global improvement was reported by 70% of physiotherapy participants (51/73) at 12 weeks and by 59% (43/73) at 24 weeks. Similarly, global improvement was reported by 72% of placebo participants (48/67) at 12 weeks and by 49% (33/67) at 24 weeks (all p>0.05). CONCLUSIONS: The physiotherapy programme tested in this trial was no more effective than regular contact with a therapist at reducing pain and disability. PMID- 15897311 TI - Pain and high sensitivity C reactive protein in patients with chronic low back pain and acute sciatic pain. AB - BACKGROUND: The severity of pain from musculoskeletal disorders might be associated with high sensitivity C reactive protein (hsCRP), a sensitive marker of low grade systemic inflammation. OBJECTIVE: To study the association between pain as assessed by a visual analogue scale (VAS) and hsCRP in patients with chronic low back pain and acute sciatic pain. METHODS: Information on pain severity, determinants of hsCRP, and hsCRP values were obtained prospectively at up to 10 time points during six months in 72 consecutive patients (mean age 43.3 years; 59.7% female): 41 with chronic low back pain and 31 with acute sciatic pain. The association between severity of pain and raised (highest quartile) hsCRP values at any time point was estimated by multivariable logistic regression using generalised estimating equations to adjust odds ratios (OR) and their confidence intervals (CI) for intraindividual dependence of measurements. RESULTS: Mean intensity of pain (VAS 0-10) at baseline was 4.9 and 5.5 in patients with chronic low back and acute sciatic pain, respectively. Highest v lowest tertile of average intensity of pain during the last 24 hours was associated with increased hsCRP levels among patients with acute sciatic pain (adjusted OR = 3.4 (95% CI, 1.1 to 10), but not in patients with chronic low back pain (adjusted OR = 0.87 (0.25 to 3.0)). CONCLUSIONS: Mean intensity of pain during the previous 24 hours as assessed by VAS was independently associated with high levels of hsCRP in patients with acute sciatic pain but not in those with chronic low back pain. PMID- 15897312 TI - Disseminated intravascular coagulation in catastrophic antiphospholipid syndrome: clinical and haematological characteristics of 23 patients. AB - BACKGROUND: Disseminated intravascular coagulation (DIC) is an acquired syndrome characterised by formation of microthrombi and fibrin deposition in the microvasculature. The catastrophic antiphospholipid syndrome (APS) is characterised by multiorgan thrombosis, mainly involving small vessels. A broad spectrum of disorders may develop DIC features; however, the catastrophic APS has not previously been recognised as a cause of DIC. OBJECTIVE: To analyse the clinical and laboratory characteristics of catastrophic APS patients with DIC features. METHODS: The web site based international registry of patients with catastrophic APS (CAPS registry) (http://www.med.ub.es/MIMMUN/FORUM/CAPS.HTM) was analysed and the cases with DIC features selected. RESULTS: In 173 patients with catastrophic APS, 23 (13%) were found with DIC features. The clinical and immunological characteristics were similar in catastrophic APS patients with and without DIC features; a significant difference was found only in the prevalence of thrombocytopenia (100% in patients with DIC features v 59% in those without DIC features). CONCLUSIONS: DIC features are not rare in catastrophic APS, supporting the need for systematic screening of antiphospholipid antibodies in all patients with DIC features without precipitating factors. The presence of DIC features in the context of an APS makes it imperative to rule out the catastrophic variant of this syndrome. PMID- 15897313 TI - Single nucleotide polymorphisms in the gene encoding the major histocompatibility complex class II transactivator (CIITA) in systemic lupus erythematosus. AB - BACKGROUND: The major histocompatibility complex (MHC) class II transactivator (CIITA) is a master switch of antigen presentation and activates expression of the MHC II gene. Insufficient up regulation of MHC class II molecules is reported to be one of the major immunological mechanisms in systemic lupus erythematosus (SLE). OBJECTIVE: To examine the association between single nucleotide polymorphisms (SNPs) in the human CIITA gene (MHC2TA) and SLE. METHODS: Promoters and coding regions of MHC2TA were evaluated for polymorphisms in 100 patients with SLE and 100 healthy donors. Eight oligonucleotide primer sets that covered the coding region and each promoter region were used for genomic analysis of SNPs. RESULTS: Allele frequencies of previously reported SNPs did not differ between healthy donors and patients with SLE. Additionally, a new polymorphism in an intronic region at nt 485 (A-->A/G) was identified, which is close to the polymorphism at nt 474 that has been associated with one of the disease causing CIITA cDNA mutations in bare lymphocyte syndrome. This SNP was found in 11% of patients with SLE and in 3% of healthy donors, suggesting it may have a role in the pathogenesis of SLE. CONCLUSIONS: A newly identified polymorphism in an intronic region at nt 485 (A-->A/G) may have an important role in the pathogenesis of SLE. PMID- 15897314 TI - Value of C reactive protein in the assessment of erosive osteoarthritis of the hand. AB - OBJECTIVE: To investigate the value of serum C reactive protein (CRP) as a marker of erosive osteoarthritis (EOA) of the hand. METHODS: Ninety eight patients, 67 with EOA and 31 with non-EOA of the hand, were included in the study and analysed for radiographic score (RS), number of erosions, and joint count (JC) at clinical observation and at bone scintigraphy. CRP was assayed in a serum sample by a highly sensitive immunonephelometric method. RESULTS: The median (interquartile range) CRP level was 4.7 (2.4-6.9) mg/l in the EOA and 2.1 (0.5-4.9) mg/l in the non-EOA group (p = 0.001). In all patients, CRP correlated with RS (r(s) = 0.43, p<0.001), and mainly with JC at clinical observation (r(s) = 0.72, p<0.001) and at bone scintigraphy (r(s) = 0.47, p<0.001). The correlation of CRP with RS and JC was confirmed at clinical observation and at bone scintigraphy in the EOA subgroup, but only with JC at clinical observation in the non-EOA subgroup. CONCLUSIONS: CRP levels are higher in EOA than in non-EOA patients. These levels probably reflect the disease activity of EOA, as suggested by correlations between CRP and JC at clinical observation and at bone scintigraphy. PMID- 15897315 TI - Peripheral ulcerative keratitis in rheumatoid arthritis: successful use of intravenous cyclophosphamide and comparison of clinical and serological characteristics. PMID- 15897316 TI - Therapeutic use of infliximab in sight threatening uveitis: retrospective analysis of efficacy, safety, and limiting factors. PMID- 15897317 TI - Systemic mastocytosis: a rare cause of osteoporosis and its response to bisphosphonate treatment. PMID- 15897318 TI - Frostbite arthritis. PMID- 15897319 TI - Methotrexate related adverse effects in patients with RA. PMID- 15897320 TI - Clues to occludin. Focus on "Knockdown of occludin expression leads to diverse phenotypic alterations in epithelial cells". PMID- 15897321 TI - Impact on fatty acid metabolism and differential localization of FATP1 and FAT/CD36 proteins delivered in cultured human muscle cells. AB - We compared the intracellular distribution and regulatory role of fatty acid transporter protein (FATP1) and fatty acid translocase (FAT/CD36) on muscle cell fatty acid metabolism. With the use of adenoviruses, FATP1 and FAT genes were delivered to primary cultured human muscle cells. FATP1 and FAT moderately enhanced palmitate and oleate transport evenly at concentrations of 0.05, 0.5, and 1 mM. Long-term (16 h) consumption of palmitate and oleate from the media, and particularly incorporation into triacylglyceride (TAG), was stimulated equivalently by FATP1 and FAT at all fatty acid concentrations tested. In contrast, long-term CO(2) production was reduced by FATP1 and FAT at all doses of palmitate and at the lower concentrations of oleate. Neither FATP1 nor FAT markedly altered the production of acid-soluble metabolic intermediates from palmitate or oleate. The intracellular localization of fusion constructs of FATP1 and FAT with enhanced green fluorescent protein (EGFP) was examined. Independently of fatty acid treatment, FATPGFP was observed throughout the cytosol in a reticular pattern and concentrated in the perinuclear region, partly overlapping with the Golgi marker GM-130. FATGFP was found in the extracellular membrane and in cytosolic vesicles not coincident with GM-130. Neither FATP1 nor FAT proteins colocalized with lipid droplets in oleate-treated cells. We conclude that whereas FAT is localized on the extracellular membrane, FATP1 is active in the cytosol and imports fatty acids into myotubes. Overall, both FATP1 and FAT stimulated transport and consumption of palmitate and oleate, which they channeled away from complete oxidation and toward TAG synthesis. PMID- 15897322 TI - A possible role for membrane depolarization in epithelial wound healing. AB - Linear narrow wounds produced on cultured bovine corneal endothelial monolayers heal by actin cable formation at the wound border and lamellar crawling of cells into the injured area. We report the novel finding that membrane potential depolarization occurs at the leading edge of wounds and gradually extends inward toward the neighboring cells. We have determined that the replacement of extracellular Na(+) by choline and the incorporation of phenamil, an inhibitor of the epithelial Na(+) channel (ENaC), provoke a decrease in the actin cable and depolarization areas and in the lamellar activity of the wound edges. To the contrary, extracellular Li(+) can successfully replace Na(+) in the determination of the depolarization and cytoskeletal responses. This finding supports the idea that membrane depolarization, not the increase in intracellular Na(+) concentration, is responsible for the formation of the actin cable, a result that is in agreement with previous evidence showing that nonspecific depolarization of the plasma membrane potential (PMP) of epithelial cells may promote characteristic cytoskeletal rearrangements per se (Chifflet S, Hernandez JA, Grasso S, and Cirillo A. Exp Cell Res 282: 1-13, 2003). We suggest that spontaneous depolarization of the PMP of the cells at the wound borders determined by a rise in the ENaC activity of these cells constitutes an additional factor in the intermediate cellular processes leading to wound healing in some epithelia. PMID- 15897323 TI - Triplet nucleotide removal at random positions in a target gene: the tolerance of TEM-1 beta-lactamase to an amino acid deletion. AB - The deletion of amino acids is one of the evolutionary mechanisms by which nature adapts the function of proteins. A simple method has been developed that mimics this event in vitro by introducing a deletion of exactly three nucleotides at random positions in a target gene. The method involved the engineering of the mini-Mu transposon to introduce a recognition sequence for the restriction enzyme MlyI. The new transposon, MuDel, was capable of efficient insertion into a target DNA sequence. To determine the efficacy of the method, the bla gene that encodes the TEM-1 beta-lactamase was used as the target and a small library containing 22 different sequence variants was created. Of these 22 variants, 8 were identified that conferred resistance to ampicillin on Escherichia coli. Each of the TEM-1 variants possessed a distinct ampicillin minimum inhibitory concentration, ranging from 500 to >10,000 microg/ml. Sequence analysis revealed that active TEM 1 variants contained deletions not just in loops but also helices, and included regions known to be involved in catalysis, antibiotic resistance and inhibitor binding. This new technology is transferable to most genes, permitting an extensive analysis of deletion mutations on protein function. PMID- 15897324 TI - Localization of spermine binding sites in 23S rRNA by photoaffinity labeling: parsing the spermine contribution to ribosomal 50S subunit functions. AB - Polyamine binding to 23S rRNA was investigated, using a photoaffinity labeling approach. This was based on the covalent binding of a photoreactive analog of spermine, N1-azidobenzamidino (ABA)-spermine, to Escherichia coli ribosomes or naked 23S rRNA under mild irradiation conditions. The cross-linking sites of ABA spermine in 23S rRNA were determined by RNase H digestion and primer-extension analysis. Domains I, II, IV and V in naked 23S rRNA were identified as discrete regions of preferred cross-linking. When 50S ribosomal subunits were targeted, the interaction of the photoprobe with the above 23S rRNA domains was elevated, except for helix H38 in domain II whose susceptibility to cross-linking was greatly reduced. In addition, cross-linking sites were identified in domains III and VI. Association of 30S with 50S subunits, poly(U), tRNA(Phe) and AcPhe-tRNA to form a post-translocation complex further altered the cross-linking, in particular to helices H11-H13, H21, H63, H80, H84, H90 and H97. Poly(U) programmed 70S ribosomes, reconstituted from photolabeled 50S subunits and untreated 30S subunits, bound AcPhe-tRNA in a similar fashion to native ribosomes. However, they exhibited higher reactivity toward puromycin and enhanced tRNA-translocation efficiency. These results suggest an essential role for polyamines in the structural and functional integrity of the large ribosomal subunit. PMID- 15897325 TI - Selection of novel mediators of E2F1-induced apoptosis through retroviral expression of an antisense cDNA library. AB - The E2F1 transcription factor is an essential mediator of p53-dependent and p53 independent apoptosis as part of an anti-tumour safeguard mechanism. In this study, a functional so-called technical knockout (TKO) approach was applied to Saos-2ERE2F1 cells that conditionally activate E2F1 by the addition of 4 hydroxytamoxifen to search for p53-independent pro-apoptotic E2F1 targets. The approach was based on random inactivation of genes after retroviral transfer of an antisense cDNA library enriched of E2F1-induced genes, followed by the selection of Saos-2ERE2F1 cells that survive in the presence of the apoptotic stimulus. We identified 13 novel E2F1 target genes encoding proteins of known cellular function, including apoptosis and RNA binding. FACS analysis revealed that E2F1-induced apoptosis was significantly attenuated in cell clones containing the antisense cDNA fragments of these genes, demonstrating their participation in E2F1 death pathways. Moreover, inactivation of the target genes resulted in a clear increase of cell viability (>80%) in response to E2F1 activation compared with controls (approximately 30%). Four genes showed an increase in expression intensity in the presence of cycloheximide, suggesting a direct effect of E2F1 on gene transcription, whereas one gene was identified as an indirect target. Our data provide new insight in the regulation of E2F1 induced apoptosis. PMID- 15897326 TI - Can statin therapy alter the natural history of bicuspid aortic valves? PMID- 15897327 TI - Nonexcitatory electrical signals for enhancing ventricular contractility: rationale and initial investigations of an experimental treatment for heart failure. PMID- 15897328 TI - Bone marrow stem cell transplantation for cardiac repair. AB - Cardiomyocytes respond to physiological or pathological stress only by hypertrophy and not by an increase in the number of functioning cardiomyocytes. However, recent evidence suggests that adult cardiomyocytes have the ability, albeit limited, to divide to compensate for the cardiomyocyte loss in the event of myocardial injury. Similarly, the presence of stem cells in the myocardium is a good omen. Their activation to participate in the repair process is, however, hindered by some as-yet-undetermined biological impediments. The rationale behind the use of adult stem cell transplantation is to supplement the inadequacies of the intrinsic repair mechanism of the heart and compensate for the cardiomyocyte loss in the event of injury. Various cell types including embryonic, fetal, and adult cardiomyocytes, smooth muscle cells, and stable cell lines have been used to augment the declining cardiomyocyte number and cardiac function. More recently, the focus has been shifted to the use of autologous skeletal myoblasts and bone marrow-derived stem cells. This review is a synopsis of some interesting aspects of the fast-emerging field of bone marrow-derived stem cell therapy for cardiac repair. PMID- 15897329 TI - Catheter-based antegrade intracoronary viral gene delivery with coronary venous blockade. AB - The purpose of this study is to evaluate the feasibility of percutaneous antegrade myocardial gene transfer (PAMGT). A consistent and safe technique for in vivo gene transfer is required for clinical application of myocardial gene therapy. PAMGT with concomitant coronary venous blockade was performed in 12 swine. The myocardium was preconditioned with 1 min of occlusion of the left anterior descending and left circumflex arteries. The anterior interventricular vein was occluded during left anterior descending artery delivery, and the great cardiac vein at the entrance of the middle cardiac vein was occluded during left circumflex artery delivery. With arterial and venous balloons inflated (3 min) and after adenosine (25 mug) injection, PAMGT was performed by antegrade injection of an adenoviral solution (1 ml of 10(11) plaque-forming units in each coronary artery) carrying beta-galactosidase or saline through the center lumen of the angioplasty balloon. In one set of animals, PAMGT was performed with selective coronary vein blockade (n = 9); in another set of animals, PAMGT was performed without coronary vein blockade (n = 5). At 1 wk after gene delivery, the animals were killed. Quantitative beta-galactosidase analysis was performed in the left and right ventricular walls. PAMGT was successfully performed in all animals with and without concomitant occlusion of the coronary veins. Quantitative beta-galactosidase analysis showed that PAMGT with coronary blockade was superior to PAMGT without coronary blockade. beta-Galactosidase activity increased significantly in the beta-galactosidase group compared with the saline group: 1.34 +/- 0.18 vs. 0.81 +/- 0.1 ng (P 50% and increased elastin and fibrillin-1 deposition by >3- and 4-fold respectively (histochemistry and immunohistochemistry). Fibrillin-1 gene expression was increased 5-fold (real time reverse-transcriptase polymerase chain reaction). Ramiprilat also reduced gene and protein (Western) expression of both matrix metalloproteinase (MMP)-2 and MMP-3. In conclusion, ramipril promoted an elastogenic matrix profile that may contribute to the observed clinical reduction in large-artery stiffness and carotid pressure augmentation, which occurred independently of mean arterial blood pressure reduction in patients with peripheral arterial disease. PMID- 15897363 TI - Dual ECE/NEP inhibition on cardiac and neurohumoral function during the transition from hypertrophy to heart failure in rats. AB - CGS 26303 is a vasopeptidase inhibitor that simultaneously inhibits endothelin converting enzyme (ECE) and neutral endopeptidase (NEP). We compared the effects of chronic treatment with CGS 26303 to the selective inhibition of angiotensin converting enzyme (ACE) and NEP during the transition from left ventricular hypertrophy (LVH) to congestive heart failure (CHF) in hypertensive rats. LV geometry and function were assessed in Dahl salt-sensitive rats placed on a high salt diet from age 6 weeks (hypertensive rats) and in control rats fed a low-salt diet. The hypertensive rats were randomized into groups that received no treatment or were treated with an ACE inhibitor (temocapril), an ECE/NEP inhibitor (CGS 26303), or a NEP inhibitor (CGS 24592) from the LVH stage (11 weeks) to the CHF stage (17 weeks). All treatments decreased the systolic blood pressure equally and significantly improved LV fractional shortening. Both temocapril and CGS 26303 ameliorated LV perivascular fibrosis, reduced mRNA levels of types I and III collagen, and decreased the heart weight/body weight ratio. CHF rats had increased plasma ET-1 levels compared with control rats. Only CGS 26303 reduced ET-1 to normal levels. ET-1 levels were found to correlate with heart/body weight, right ventricle/body weight and perivascular fibrosis ratios. During the transition to CHF, CGS 26303 produces effects that are comparable to temocapril and superior to CGS 24592. The beneficial effects of CGS 26303 are likely caused in part by the greater reduction of plasma ET-1. Dual ECE/NEP inhibitor may provide a new strategy for the treatment of human heart failure. PMID- 15897364 TI - Can we afford crude estimates of central pulse pressure? PMID- 15897367 TI - The role of the alpha1B-adrenergic receptor in vascular structure and function. PMID- 15897368 TI - Management of prehypertension. PMID- 15897369 TI - Long-term impact of systolic blood pressure and glycemia on the development of microalbuminuria in essential hypertension. AB - The objective was to assess the temporal impact of factors related to the development of microalbuminuria during the follow-up of young adult normoalbuminurics with high-normal blood pressure or at stage 1 of essential hypertension. Prospective follow-up was conducted on 245 normoalbuminuric hypertensive subjects (mean age 40.9 years; 134 men; blood pressure 139.7/88.6 mm Hg; body mass index 28.5 kg/m2) never treated previously with antihypertensive drugs, with yearly urinary albumin excretion measurements, until the development of microalbuminuria. After enrollment, patients were placed on usual care including nonpharmacological treatment or with an antihypertensive drug regime to achieve a blood pressure of <135/85 mm Hg. Thirty subjects (12.2%) developed microalbuminuria after a mean follow-up of 29.9 months (range 12 to 144 months), 2.5 per 100 patients per year. Baseline urinary albumin excretion (hazard ratio, 1.07; P=0.006) and systolic blood pressure during the follow-up (hazard ratio, 1.03; P=0.008) were independent factors related to the follow-up urinary albumin excretion in a Cox proportional hazard model. A significant increase in the risk of developing microalbuminuria for urinary albumin excretion at baseline >15 mg per 24-hour systolic blood pressure >139 mm Hg and a positive trend in fasting glucose were observed in the univariate analyses. However, in the multivariate analysis, only the baseline urinary albumin excretion and the trend of fasting glucose were independently related to the risk of developing microalbuminuria. In mild hypertensives, the development of microalbuminuria was linked to insufficient blood pressure control and to a progressive increment of glucose values. PMID- 15897370 TI - Kidney in early atherosclerosis. AB - Atherosclerosis represents one of the major causes of premature death in the United States today, and it is frequently associated with, exacerbates, and is aggravated by chronic kidney disease (CKD). Atherosclerosis integrates the response to a number of insults, and consequently, the accelerated atherosclerosis found in CKD patients is associated with activation of a variety of humoral and tissue mechanisms. Hypertension, diabetes, dyslipidemia, obesity, metabolic syndrome, and additional nontraditional risk factors can damage the kidney directly and by promoting intrarenal atherogenesis, even in the absence of obstructive lesions in the renal artery. Evidence indicates that increased oxidative stress and inflammation may mediate a large part of the effects of risk factors on the kidney. In turn, progressive deterioration of renal function in CKD may lead to dyslipidemia or accumulation of uremic toxins, which can induce production of free radicals and activate proinflammatory and fibrogenic factors, leading to vascular endothelial cell dysfunction and injury, and favoring development of atherosclerosis. Therefore, the kidney can be a villain or a victim during atherogenesis. The purpose of this review is to provide new insights into the mechanisms by which atherogenic factors may instigate early renal injury. PMID- 15897371 TI - Effect of body mass index changes between ages 5 and 14 on blood pressure at age 14: findings from a birth cohort study. AB - Weight reduction in clinical populations of severely obese children has been shown to have beneficial effects on blood pressure, but little is known about the effect of weight gain among children in the general population. This study compares the mean blood pressure at 14 years of age with the change in overweight status between ages 5 and 14. Information from 2794 children born in Brisbane, Australia, and who were followed up since birth and had body mass index (BMI) and blood pressure measurements at ages 5 and 14 were used. Systolic and diastolic blood pressure at age 14 was the main outcomes and different patterns of change in BMI from age 5 to 14 were the main exposure. Those who changed from being overweight at age 5 to having normal BMI at age 14 had similar mean blood pressures to those who had a normal BMI at both time points: age- and sex adjusted mean difference in systolic blood pressure 1.54 (-0.38, 3.45) mm Hg and in diastolic blood pressure 0.43 (-0.95, 1.81) mm Hg. In contrast, those who were overweight at both ages or who had a normal BMI at age 5 and were overweight at age 14 had higher blood pressure at age 14 than those who had a normal BMI at both times. These effects were independent of a range of potential confounding factors. Our findings suggest that programs that successfully result in children changing from overweight to normal-BMI status for their age may have important beneficial effects on subsequent blood pressure. PMID- 15897372 TI - Hyperleptinemia: protecting the heart from lipid overload. AB - In this review, we attempt to deduce teleologically the physiological mission of leptin. Because overnutrition and diet-induced obesity are the only known causes of hyperleptinemia, we contrast the differences in overnutrition in normally leptinized rodents, in which the added lipids are confined to adipocytes, with those of unleptinized rodents, in which the added lipids are distributed in liver, pancreatic islets, and heart and skeletal muscle, causing organ dysfunction and cell death with a disease cluster resembling metabolic syndrome. We focus here on lipid-induced cardiac dysfunction and the remarkable ability of hyperleptinemia to prevent it. We conclude that the hyperleptinemia of overnutrition prevents the ectopic lipid deposition by: (1) acting on hypothalamic appetite centers to limit the caloric surplus to fit the available adipocyte storage capacity and, (2) upregulating of fatty acid oxidation and downregulating lipogenesis in peripheral tissues to minimize ectopic lipid deposition. The causes of failure of this system and its clinical consequences are discussed. PMID- 15897373 TI - Effects of weight loss in overweight/obese individuals and long-term hypertension outcomes: a systematic review. AB - Many studies have assessed short-term effects of weight loss on blood pressure, whereas little attention has been paid to long-term effects. We conducted a systematic review to evaluate the long-term effects of weight loss on hypertension outcome measures in adults using literature published from 1966 to 2001. All prospective studies and trials, performed on participants with body mass index of > or =28 kg/m2 with a follow-up of >2 years and weight changes recorded, were included. The data from these studies were used to model the long term effects on blood pressure. Previous reviews on shorter-term studies indicate a 1:1 drop in blood pressure (mm Hg) with weight loss (kilograms). Our findings, based on studies with follow-up of > or =2 years, demonstrate blood pressure decreases less than this after weight loss. The surgical intervention studies exhibited huge weight losses with undramatic blood pressures changes. When surgical interventions are excluded, the models suggest that for 10 kg weight loss, decreases of 4.6 mm Hg and 6.0 mm Hg in diastolic and systolic blood pressure, respectively, may be expected, about half of that predicted from the short-term trials. Initial blood pressure, the length of follow-up, medication changes, and physiological restrictions may contribute to this reduced effect in the long-term studies. Extrapolation of short-term blood pressure changes with weight loss to the longer term is potentially misleading. The weight/hypertension relationship is complex and needs well-conducted studies with long-term follow-up to examine the effects of weight loss on hypertension outcomes. PMID- 15897374 TI - Intralesional fluorouracil injection in infantile digital fibromatosis. PMID- 15897375 TI - Extensive aplasia cutis congenita and the risk of sagittal sinus thrombosis. PMID- 15897376 TI - Isotretinoin therapy and mood changes in adolescents with moderate to severe acne: a cohort study. AB - OBJECTIVE: To determine whether patients with moderate to severe acne who were treated with isotretinoin experienced significant increases in depressive symptoms over a 3- to 4-month period compared with patients who received conservative acne therapy. DESIGN: Cohort study. SETTING: Hospital-affiliated and community-based clinics in St Louis, Mo. PARTICIPANTS: One hundred thirty-two subjects aged 12 to 19 years with moderate to severe acne. MAIN OUTCOME MEASURES: Depressive symptoms were assessed using the Center for Epidemiological Studies Depression Scale (CES-D), a standardized self-reported instrument. Mean CES-D scores were compared between treatment groups, as were the prevalence and incidence of scores suggestive of clinically significant depression (CES-D score >16). RESULTS: A total of 101 subjects completed the study. At follow-up, CES-D scores (adjusted for baseline CES-D score and sex of patient) suggestive of clinically significant depression were no more prevalent in the isotretinoin group than in the conservative therapy group. Similarly, the incidence (new onset) of depressive symptoms suggestive of clinical significance also was not significantly different between the treatment groups. CONCLUSIONS: The use of isotretinoin in the treatment of moderate-severe acne in adolescents did not increase symptoms of depression. On the contrary, treatment of acne either with conservative therapy or with isotretinoin was associated with a decrease in depressive symptoms. PMID- 15897377 TI - Trends in adherence to a revised risk management program designed to decrease or eliminate isotretinoin-exposed pregnancies: evaluation of the accutane SMART program. AB - OBJECTIVE: To review adherence to selected procedures outlined in the System to Manage Accutane-Related Teratogenicity (SMART) program during the first year of implementation vs the procedures in effect in the year prior to initiation of the SMART program. DESIGN: Observational. SETTING: A novel pharmacy compliance survey and an ongoing, voluntary survey. PATIENTS: Female recipients of isotretinoin. INTERVENTION: In April 2002, Hoffmann-La Roche Inc, Nutley, NJ, manufacturer of Accutane brand isotretinoin and at that time the sole source of isotretinoin, revised earlier guidelines and instituted the SMART risk management program, which included the use of qualification stickers to affix to all prescriptions for Accutane to indicate, among other things, a negative pregnancy test just before the prescription was written. The goal of the SMART program was to decrease or eliminate isotretinoin-exposed pregnancies. MAIN OUTCOME MEASURES: Use and completion of prescription qualification stickers; changes in pretherapy pregnancy testing and birth control use. RESULTS: The results of the pharmacy compliance survey indicated high (>90%) use of prescription qualification stickers. Results of the patient survey suggested that 9% of prescription qualification stickers within the observed user cohort were issued without a pregnancy test. Furthermore, the pregnancy rate for patients participating in the survey was similar to that reported for cohorts recruited before the SMART program. CONCLUSIONS: The usefulness of the results derived from 2 surveys designed to evaluate the SMART program is limited by the lack of reliability and validity of the survey instruments and by questionable generalizability to all female recipients of isotretinoin. The presence of a qualification sticker may not have an impact on pregnancy testing or compliance with effective birth control behavior as outlined in the SMART program. PMID- 15897378 TI - General anesthesia for pediatric dermatologic procedures: risks and complications. AB - OBJECTIVE: To assess the safety and adverse events associated with the use of general anesthesia in children undergoing elective dermatologic procedures. DESIGN: A multicenter retrospective review. SETTING: Children's Hospital and Health Center, San Diego, Calif, and Northwestern University School of Medicine, Chicago, Ill. PATIENTS: The study population comprised 269 children and adolescents ranging in age from 2 months to 18 years (881 procedures performed by 6 pediatric dermatologic and laser surgeons). MAIN OUTCOME MEASURES: The risk of an adverse event occurring during general anesthesia for pediatric dermatologic procedures. RESULTS: The risk of general anesthesia in elective pediatric dermatologic procedures was low: 90% of patients experienced no clinically relevant complications. The most common clinically relevant adverse effect of general anesthesia was perioperative nausea and emesis, which was noted in 4% of patients. There were no serious life-threatening events noted, and the mortality rate was 0%. CONCLUSION: The use of general anesthesia for dermatologic procedures in a children's hospital setting appears safe, with a low rate of complications. PMID- 15897379 TI - The prevalence of common acquired melanocytic nevi and the relationship with skin type characteristics and sun exposure among children in Lithuania. AB - OBJECTIVE: To evaluate the prevalence of common acquired melanocytic nevi and its relationship with pigmentary characteristics and severe sunburns in children. DESIGN: Cross-sectional study. SETTING: Kaunas city, Lithuania (latitude of 55 degrees 55 minutes). PATIENTS: A random sample of 484 children aged 1 to 2 years, 4 to 5 years, and 9 to 10 years and adolescents aged 14 to 15 years. INTERVENTIONS: A questionnaire that provided information about the history of sunbathing and skin type. MAIN OUTCOME MEASURES: Atypical melanocytic nevi were defined according to the clinical criteria of the ABCDE rule. RESULTS: The median number of all common acquired melanocytic nevi was 12 in boys and 11 in girls; the median number of melanocytic nevi 2 mm or larger in boys and girls was 4. Twenty-seven percent of children experienced severe sunburns more than once. After adjustment for age and sex, it was found that children who had severe sunburns in summer and skin type I had a higher density of all melanocytic nevi and melanocytic nevi 2 mm or larger. The prevalence of atypical melanocytic nevi was 7% in all children and was age dependent (age 4-5 years, 1%; 9-10 years, 4%; 14-15 years, 16%). Three percent of children had congenital melanocytic nevi. CONCLUSIONS: The total number of common acquired melanocytic nevi in children increased with age. There was a positive association between severe sunburns, the tendency of the skin to burn, and the number of all melanocytic nevi and nevi 2 mm or larger. PMID- 15897380 TI - Intralesional immunotherapy of warts with mumps, Candida, and Trichophyton skin test antigens: a single-blinded, randomized, and controlled trial. AB - BACKGROUND: Warts occur commonly in humans. Destructive modalities are generally the first physician-administered therapy. Other treatment options include immunotherapy. Intralesional immunotherapy using mumps, Candida, or Trichophyton skin test antigens has proved efficacy in the treatment of warts. OBJECTIVES: To determine rates of wart resolution in response to injection of antigen alone, antigen plus interferon alfa-2b, interferon alfa-2b alone, and normal saline; and to compare response according to viral type, major histocompatibility complex antigens, and peripheral blood mononuclear cell proliferation to autologous human papillomavirus antigen before and after injection. DESIGN: Randomized, single blinded, placebo-controlled, clinical trial. SETTING: Medical school-based dermatology department. PATIENTS: Two hundred thirty-three patients clinically diagnosed as having 1 or more warts. Main Outcome Measure Clinical resolution of warts in response to intralesional immunotherapy. RESULTS: Responders were observed in all treatment arms, but were significantly more likely to have received antigen (P<.001). Resolution of distant untreated warts was observed, and was significantly more likely in subjects receiving antigen (P<.001). Interferon did not significantly enhance the response rate (P = .20) and did not differ from normal saline (P = .65). No viral type or major histocompatibility complex antigen correlated with response or lack of response (P>.99 and P = .86, respectively). A positive peripheral blood mononuclear cell proliferation assay result (2 times pretreatment levels) was significantly more likely among responders (P = .002). While there was no significant difference in response based on sex (P = .56), older subjects (>40 years) were less likely to respond (P = .01). CONCLUSIONS: Intralesional immunotherapy using injection of Candida, mumps, or Trichophyton skin test antigens is an effective treatment for warts, as indicated by significantly higher response rates and distant response rates in subjects receiving antigen. Viral type and major histocompatibility complex antigens did not seem to influence treatment response. Response is accompanied by proliferation of peripheral blood mononuclear cells to human papillomavirus antigens, suggesting that a human papillomavirus-directed cell-mediated immune response plays a role in wart resolution. PMID- 15897381 TI - Comparison of colchicine, dapsone, triamcinolone, and diphenhydramine therapy for the treatment of brown recluse spider envenomation: a double-blind, controlled study in a rabbit model. AB - OBJECTIVE: To compare the efficacy of dapsone, diphenhydramine, colchicine, and intralesional triamcinolone in the treatment of brown spider bites. We used a purified venom that reproducibly produces a large eschar. To mimic real-life circumstances, all agents were administered following a 2-hour delay after envenomation. The animals were evaluated for the presence of coagulopathy to determine if the incidence of systemic findings correlated with the type of treatment. DESIGN AND SETTING: In a research laboratory, 60 New Zealand white rabbits each received an intradermal injection of 20 microg of purified Loxosceles reclusa venom. The rabbits were divided into 5 groups of 12; a control group and 4 groups treated with a drug (either colchicine, triamcinolone, diphenhydramine, or dapsone). Measured end points included maximum eschar size as well as histologic grading of depth, inflammation, and thrombosis. INTERVENTIONS: Treatment with colchicine, triamcinolone, diphenhydramine, or dapsone. MAIN OUTCOME MEASURES: Maximum eschar size as well as histologic grading of depth, inflammation, and thrombosis. RESULTS: There was no significant difference with respect to eschar size (1-way analysis of variance, P = .003). There was no significant difference between any treatment with respect to presence or absence of ulcer, necrosis, large vessel vasculitis, or small vessel vasculitis. The only outcome of significance was that triamcinolone offered protection from thrombosis (chi2 likelihood ratio, P = .04). We also noted evidence of coagulopathy in all of the envenomated animals. The rabbits had grossly elevated activated partial thromboplastin time results, which were corrected with 1:1 mixing with normal rabbit plasma, suggesting an acquired factor deficiency. We did not detect an individual factor deficiency or a lupus anticoagulant. CONCLUSIONS: In a rabbit model, none of the agents tested (dapsone, diphenhydramine, colchicine, and intralesional triamcinolone) had an effect on eschar size. Triamcinolone appeared to offer some protection against histologic evidence of thrombosis, but this protection did not translate into a difference in clinical outcome. All animals developed evidence of coagulopathy, regardless of treatment. The coagulopathy could be corrected by fresh rabbit plasma, suggesting an acquired factor deficiency. PMID- 15897382 TI - Hormonal effect on psoriasis in pregnancy and post partum. AB - OBJECTIVES: To investigate prospectively how psoriasis fluctuates in pregnancy and post partum and to correlate hormone levels in pregnancy (progesterone and estrogens) with psoriatic change. DESIGN: Psoriatic body surface area (BSA) in pregnant patients with psoriasis (study group) and nonpregnant, menstruating patients with psoriasis (control group) were assessed 5 times over a year. Hormone levels (progesterone and estrogens) were measured in the study group and correlated with change in BSA. SETTING: University-affiliated obstetric and dermatology clinics. PATIENTS: Forty-seven pregnant patients in the psoriasis group and 27 nonpregnant, menstruating patients in the control group. RESULTS: During pregnancy, 55% of the patients reported improvement, 21% reported no change, and 23% reported worsening. However, post partum, only 9% of patients reported improvement, 26% reported no change, and 65% reported worsening. Psoriatic BSA decreased significantly from 10 to 20 weeks' gestation (P<.001) compared with controls, whereas BSA increased significantly by 6 weeks post partum (P = .001) compared with controls. In patients with 10% or greater psoriatic BSA who reported improvement (n = 16; mean BSA, 40%), lesions decreased by 83.8% during pregnancy. There were significant or near significant correlations between improvement in BSA and estradiol (P = .009, r = 0.648), estriol (P = .06, r = 0.491), and the ratio of estrogen to progesterone (P = .006, r = 0.671). CONCLUSION: High levels of estrogen correlated with improvement in psoriasis, whereas progesterone levels did not correlate with psoriatic change. PMID- 15897383 TI - Restrictive dermopathy associated with transposition of the great arteries and microcolon: a rare neonatal entity with new symptoms. AB - BACKGROUND: Restrictive dermopathy is a very rare autosomal recessive skin disorder. The typical pathologic findings are striking: microstomia, micrognathia, thin but very tight translucent skin that tears spontaneously, and arthrogryposis multiplex. The mechanisms behind this disease are unknown. OBSERVATIONS: We describe for the first time a newborn girl with restrictive dermopathy, transposition of the great vessels, and microcolon. She had thin shiny skin with nearly no compliance indicating restrictive dermopathy. Additional dysmorphic findings included enlarged fontanelle, hypertelorism, absent eyelashes, small pinched nose, microstomia, micrognathia, dysplastic ears, pterygium colli, dyplastic fingers and toes with upper- and partial lower-limb flexion contractures, dysplastic genitalia, and muscular hypotonia. She also had left transposition of the great artery with small atrial septal defect, bilateral hypoplasia of the first rib, and congenital stenosis of the small bowel with microcolon. CONCLUSIONS: The pathogonomic diagnostic features remain reduced dermal thickness and nearly complete absence of elastic fibers in the dermis. In mice, a defective fatty acid transport protein 4 gene (Fatp4) leads to clear signs of restrictive dermopathy by influencing the arrangement of the lipids in the epidermis. Whether the left transposition of the great artery is associated with restrictive dermopathy or represents an additional malformation of multifactorial, polygenetic, or monogenetic cause remains open. PMID- 15897384 TI - A patient with Rothmund-Thomson syndrome and all features of RAPADILINO. AB - BACKGROUND: Mutations of the human helicase gene RECQL4 have been identified in a subset of patients with Rothmund-Thomson syndrome (RTS) and in children with the diagnosis of RAPADILINO syndrome (RAdial hypoplasia/aplasia, PAtellar hypoplasia/aplasia, cleft or highly arched PAlate, DIarrhea and DIslocated joints, LIttle size [>2 SDs below the mean in height] and LImb malformation, and slender NOse and NOrmal intelligence). While many features of the 2 genetic disorders overlap, poikiloderma--a hallmark of RTS--has been described as generally absent in RAPADILINO syndrome. OBSERVATIONS: We report herein a patient with RTS who carries a truncating mutation and a newly identified missense mutation of RECQL4. The proband uniquely developed all criteria of RAPADILINO in addition to his prominent skin findings. CONCLUSIONS: Patients with RTS may possess all features of RAPADILINO. Consequently, a genetic approach to RTS and RAPADILINO could be beneficial. This approach may provide a better understanding of the wide variety of related phenotypic findings and improve prognostics. PMID- 15897385 TI - Aquagenic wrinkling of the palms in patients with cystic fibrosis homozygous for the delta F508 CFTR mutation. AB - BACKGROUND: Aquagenic wrinkling of the palms (AWP) is a rare condition characterized by the rapid and transient formation of edematous whitish plaques on the palms on exposure to water (the so-called hand-in-the-bucket sign). The changes may be asymptomatic or accompanied by pruritic or burning sensations. First described in 1974 in patients with cystic fibrosis--and still primarily reported in these patients--this condition has been previously described only in females. Specific mutations in CFTR, the gene responsible for cystic fibrosis, have not been reported previously in patients with AWP. OBSERVATIONS: We describe 2 patients with AWP, both of whom are homozygous for the delta F508 mutation in CFTR: a 17-year-old boy--the first male reported to have this condition--who has had AWP for 3 years and a 13-year-old girl who has had AWP for 6 months. CONCLUSIONS: Cystic fibrosis should be considered in patients with AWP, and patients with cystic fibrosis should be asked about symptoms of this condition. Although the etiology of AWP is unknown, the association with cystic fibrosis, and with marasmus and cyclooxygenase-2 inhibitors, suggests that exposure of the skin to abnormally high concentrations of salt may play a role in its pathogenesis. PMID- 15897387 TI - Diffuse nonepidermolytic palmoplantar keratoderma caused by a recurrent nonsense mutation in DSG1. AB - BACKGROUND: Mutations in genes coding for 2 desmosomal proteins, desmoglein 1 and desmoplakin, have been shown to cause autosomal dominant keratoderma palmoplantaris striata. OBSERVATIONS: We describe a family affected with a diffuse nonstriated form of palmoplantar keratoderma. Histopathologic examination of skin biopsy specimens disclosed cell-cell disadhesion in the suprabasal layers of the epidermis, as previously described in keratoderma palmoplantaris striata. We therefore genotyped all family members using microsatellite markers encompassing 3 keratoderma palmoplantaris striata-associated loci. Haplotype analysis suggested linkage of the disease to 18q12.1, which harbors the DSG1 gene, encoding desmoglein 1. Mutation analysis eventually led to the identification of a causative recurrent nonsense mutation in this gene. CONCLUSIONS: Mutations in DSG1 are not exclusively associated with striated palmoplantar keratoderma. The present study illustrates the efficacy of an integrative diagnostic approach to palmoplantar keratodermas involving clinical assessment, pathologic examination, microsatellite marker screening, and mutational analysis. PMID- 15897388 TI - General anesthesia in infants and children for pediatric dermatologic procedures: is it worth it? PMID- 15897389 TI - A new day for "On the Horizon". PMID- 15897390 TI - cDNA microarrays and cutaneous oncology. PMID- 15897391 TI - Necrotic ulcer on the thumb of a teenager with leukemia. PMID- 15897392 TI - Angioedema, eosinophilia, and fever. PMID- 15897393 TI - Asymptomatic preauricular nodule. PMID- 15897394 TI - Multiple painful cutaneous nodules. PMID- 15897395 TI - Relationship between headache and depression in users of isotretinoin. PMID- 15897396 TI - Do neoplastic stem cells underlie the pathogenesis of cutaneous lymphomas? PMID- 15897397 TI - Variability of dermoscopic features of tungiasis. PMID- 15897398 TI - Treatment of pyogenic granuloma by sodium tetradecyl sulfate sclerotherapy. PMID- 15897399 TI - Acquired agminated acral angioma: a novel vascular lesion. PMID- 15897400 TI - Nail dystrophy associated with a heterozygous mutation of the nude/SCID human FOXN1 (WHN) gene. PMID- 15897401 TI - Nevus sebaceous and stroke. PMID- 15897402 TI - Hyperkeratotic rim of gram-negative toe web infections. PMID- 15897403 TI - Caudal septoplasty for treatment of septal deviation: aesthetic and functional correction of the nasal base. AB - OBJECTIVES: To describe our technique in the treatment of significant caudal septal deviation; to evaluate the effectiveness of our technique of caudal septoplasty in the treatment of caudal septal deviations. DESIGN: Retrospective review of cases taken from a database of more than 2000 patients who underwent rhinoplasty performed by 1 surgeon in a private facial plastic surgery practice. RESULTS: Medical charts were reviewed to determine the rate of preoperative nasal obstruction in 59 (95%) of 62 patients as well as nasal obstruction postoperatively 11 (17%) of 62 (P < .001). Photographs were reviewed to determine the severity of caudal deviation and the postoperative result. By photographic evaluation, we found that all but 3 patients had significant improvement in their postoperative appearance. Twenty-six patients had no evidence of residual asymmetry. The rate of revision was 5 (8%) of 62 patients. CONCLUSIONS: The caudal septoplasty technique is effective, relatively easy to perform, and shows long-term reliability in correcting caudal septal deviation. In properly selected patients, the technique is effective in improving cosmesis and nasal airflow. PMID- 15897405 TI - Modulation of wound response with growth factors and platelet concentrate. PMID- 15897404 TI - Modulation of wound response and soft tissue ingrowth in synthetic and allogeneic implants with platelet concentrate. AB - OBJECTIVE: To evaluate the modulation of wound healing and soft tissue ingrowth in synthetic and allogeneic implants with platelet gel. Attempts to influence wound healing with exogenous growth factors are highly dependent on the timing and dosing of treatment. Platelet gel made from autologous platelet concentrate (PC) and activated with calcium thrombin is increasingly used to enhance healing of surgical and chronic wounds, based on the assumption that proteins found in the blood can promote healing. METHODS: Adult New Zealand white rabbits underwent phlebotomy, and the blood was used to produce nonconcentrated autologous blood clot, platelet-poor plasma (PPP), and PC for each animal. Disks of porous high density polyethylene (PHDPE) and acellular dermal graft (ADG) were implanted into each animal in a subcutaneous location. Implants of each type were treated with isotonic sodium chloride solution, PPP, PPP followed immediately with PC, or autologous blood clot (by means of manual impregnation). Animals were killed at 2, 7, 14, and 21 days after implantation. Implants were harvested with surrounding soft tissue and examined by means of light microscopy for evidence of acute and chronic inflammatory cells and vascular and fibroblast invasion. RESULTS: A platelet gel with platelet concentrations averaging 5.8 times greater than those of peripheral blood significantly improved wound healing and soft tissue ingrowth in surgically implanted grafts. Early inflammatory infiltrates were enhanced in PHDPE and ADG implants by PC and autologous blood clot compared with control implants, as evidenced by significantly increased neutrophil and macrophage counts at day 2. Compared with controls, statistically significant increases in fibroblast and endothelial cell counts were noted at day 7 in PC treated implants (fibroblasts, 61% increase [P < .001] in PHDPE implants and 52% increase [P < .001] in ADG implants; capillaries, 95% increase [P < .05] in PHDPE and 97% increase [P < .001] in ADG implants). Lymphocyte counts were increased by PC in PHDPE and ADG implants (71% [P < .001] and 100% [P < .05], respectively). There were no statistically significant differences in any cell count variables beyond 7 days. CONCLUSIONS: Treatment with PC prepared at 5 times the baseline platelet count significantly accelerated maturation of experimental wounds. By 14 days, the degree and quality of wound cellularity were equivalent among all treatment groups. Rapid wound healing was expected with this surgical model, which was chosen to observe the biological effects on early wound healing of a platelet gel in a noncompromised wound. Treatment with PC may be useful in scenarios in which enhancement and acceleration of early wound healing is desired. PMID- 15897406 TI - Topical mitomycin C in the prevention of keloid scar recurrence. AB - OBJECTIVE: To assess the ability of mitomycin C to prevent the recurrence of surgically excised keloid scars. Mitomycin C has been successfully used to prevent scar tissue formation at the site of subglottic stenosis in the field of pediatric otolaryngology. It appears that mitomycin C interferes with the ability of fibroblasts to produce a scar without causing changes in epithelialization. DESIGN: We excised keloid scars from various sites in the head and neck and then applied mitomycin C to the resected bed prior to closure of the wound at a concentration of 0.4 mg/mL for 5 minutes. All patients had multiple keloids and acted as their own control. At 1 month after the procedure both wounds were started on a regimen of triamcinolone acetonide, 40 mg/mL injections, repeated every month for 6 months. At the end of the study, photos and measurements were again taken. RESULTS: Fifteen patients (13 female and 2 male) ranging in age from 10 to 55 years enrolled in the study. No infections or nonhealing wounds were seen. There was no difference in postoperative pain. Eight patients completed the triamcinolone injections, 5 had fewer than 6 injections, and 2 patients had no steroid injections. Twelve patients completed follow-up and were evaluated for surgical complications and recurrence of the keloids at either site. Two patients had partial postoperative follow-up in person and then completed follow-up via telephone. One patient could not be found for follow-up. Four patients had recurrence of both excised lesions. Ten patients had no recurrence of their keloids at either site. CONCLUSION: Mitomycin C made no difference in the prevention of keloid recurrence after excision when topically applied. PMID- 15897407 TI - The extended columellar strut-tip graft. AB - The extended columellar strut-tip graft is a structural unit used in endonasal rhinoplasty that combines the attributes of the columellar strut and the tip graft. It is used to provide projection and contour to the nasal tip. Our goal with this study was to evaluate a 15-year experience with 155 patients who underwent rhinoplasty with the extended columellar strut-tip graft. Of these, 110 underwent secondary rhinoplasty, and 45 underwent primary rhinoplasty. There were 6 patients in the secondary rhinoplasty group who experienced complications: in 3, the graft became visible postoperatively, and 3 patients had graft placement asymmetry. These 6 patients underwent surgery in the initial years of graft development. One patient with graft edge visibility and 1 patient with graft asymmetry underwent revision surgery with satisfactory results. The extended columella strut-tip graft is a reliable method to provide nasal tip projection and contour. The successful use of the graft requires precise diagnosis and surgical technique. PMID- 15897408 TI - Postoperative chemosis after cosmetic eyelid surgery: surgical management with conjunctivoplasty. AB - Persistent conjunctival chemosis in patients who have undergone cosmetic eyelid surgery is an uncommon but aesthetically undesirable and potentially debilitating postoperative complication. We attempted to determine if a regional conjunctivoplasty can effectively reduce signs and symptoms of chronic postoperative chemosis in these patients. Three patients with iatrogenic chemosis resistant to conservative management (ie, lubrication, pressure patching, and steroid drops) and persisting for more than 6 months after the initial surgery were included. A limbal peritomy followed by subconjunctival and sub-Tenon's fascia dissection was performed in regions of clinically detectable chemosis. Patients were followed postoperatively for 3 to 9 months to assess clinical response. All patients demonstrated significant objective improvement in clinically observable chemosis as well as symptoms related to ocular surface dryness. PMID- 15897409 TI - Defects of the nasal internal lining: etiology and repair. AB - OBJECTIVES: To analyze risk factors leading to full-thickness (FT) defects, to review methods of repair, and to present guidelines for management of aggressive basal cell carcinomas (BCCs) of the nose. DESIGN: Retrospective medical chart review of patients who underwent nasal reconstruction by the Department of Otolaryngology-Head and Neck Surgery between 1996 and 2003. RESULTS: Two hundred ten patients underwent nasal reconstruction; 183 had complete medical records and were included in this study. There were 53 patients with FT nasal defects, 38 (71.7%) of which were due to BCC. Among all patients presenting with an aggressive histologic subtype of BCC, 30.1% (22/73) developed FT defects. In contrast, 14.5% (16/100) with a nonaggressive subtype had FT involvement (P < .05). CONCLUSIONS: Internal lining defects are more likely to occur from aggressive histologic subtypes of BCC (infiltrative, morpheaform, and micronodular) than nonaggressive subtypes (P < .05). For BCCs 1 to 2 cm2 located on the nasal ala, histologic subtype is a significant risk factor for resulting in a FT defect, which should influence the method of excision (direct vs Mohs micrographic surgery) and the anticipated reconstruction. Large lesions (> 2 cm2) involving the ala have a high rate of internal lining involvement independent of pathologic subtype. PMID- 15897410 TI - Aesthetic otoplasty: wedge excision of a flattened helix to create a helical curl. PMID- 15897411 TI - Treatment of periorbital rhytids with botulinum toxin type A: maximizing safety and results. AB - Botulinum toxin type A is widely used for treatment of facial rhytids, often in an "off-label" fashion. Important mechanisms of action and safety concerns for such treatments are presented along with recommendations for treatment of periorbital rhytids. PMID- 15897412 TI - Efficacy of an over-the-counter lip enhancer in lip augmentation. PMID- 15897414 TI - Consortium of otolaryngology--head and neck surgery journals to collaborate in maintenance of high ethical standards. PMID- 15897415 TI - Heritability of recurrent tonsillitis. AB - OBJECTIVE: To estimate the relative contribution of genetic and environmental effects on the variance in the liability of recurrent tonsillitis. DESIGN: Retrospective questionnaire data from a population-based cohort. SETTING: Population-based data from Norway. PARTICIPANTS: A total of 9479 Norwegian twins born between January 1, 1967, and December 31, 1979, identified through the Medical Birth Registry of Norway. Main Outcome Measure Recurrent tonsillitis. RESULTS: The lifetime prevalence of recurrent tonsillitis was 11.7% (95% confidence interval, 11.0%-12.3%), with a significant predominance of female cases. The tetrachoric correlations for monozygotic twins were 0.71 for males and 0.60 for females. For dizygotic twins, the correlations were 0.12 for males, 0.14 for females, and 0.24 for dizygotic pairs of opposite sex. Structural equation modeling indicated that genetic effects explained 62% of the variation in the liability of recurrent tonsillitis. The remaining variance was attributed to individual environmental effects. There was no evidence of sex-specific genetic effects on the liability of recurrent tonsillitis. CONCLUSION: There is evidence for a substantial genetic predisposition for recurrent tonsillitis. PMID- 15897416 TI - Investigation of the role of congenital cytomegalovirus infection in the etiology of enlarged vestibular aqueducts. AB - OBJECTIVE: To determine whether congenital cytomegalovirus (CMV) infection is an etiologic factor in the pathogenesis of enlarged vestibular aqueducts (EVA). DESIGN: Two different cohort studies. Subjects The study population comprised 19 subjects with a history of congenital CMV infection and sensorineural hearing loss (cohort 1); 39 subjects with nonsyndromic EVA and their unaffected mothers (cohort 2); and 16 control subjects with EVA associated with Pendred syndrome and bi-allelic mutations of the SLC26A4 gene and their unaffected mothers. RESULTS: In cohort 1, we detected EVA in 0 of 19 subjects with congenital CMV infection and sensorineural hearing loss. In cohort 2, anti-CMV serologic profiles were consistent with possible congenital CMV infection in 10 (26%) of 39 subjects with nonsyndromic EVA and 6 (38%) of 16 control subjects with Pendred syndrome (P = .52). These seroprevalence rates are similar to those expected in the general population (40%). CONCLUSION: In spite of their auditory phenotypic similarities, congenital CMV infection is not a significant factor in the etiology of EVA. PMID- 15897417 TI - Fetal surgery in otolaryngology: a new era in the diagnosis and management of fetal airway obstruction because of advances in prenatal imaging. AB - OBJECTIVE: To evaluate the efficacy, safety, and outcome of prenatal imaging and fetal surgery in the diagnosis and management of fetal airway obstruction caused by cervical teratoma or lymphatic malformation. SETTING: Tertiary care medical center. Patients A retrospective study of all consecutive fetal patients with cervical teratoma or lymphatic malformation between January 2001 and December 2003. RESULTS: The indication was potential airway obstruction due to a fetal neck mass in 8 patients. Prenatal images were obtained by ultrasonography and magnetic resonance imaging, and were consistent with teratoma in 4 patients. The mean cervical mass was 8.3 x 7.3 x 6.7 cm, with airway obstruction suspected in all 4 patients. All 4 patients were successfully delivered by ex utero intrapartum treatment, during which 3 newborns required tracheotomy and 1 was successfully intubated. Prenatal images were consistent with lymphatic malformation in the remaining 4 patients. The mean cervical mass was 4.6 x 4.4 x 3.4 cm. There was no indication of airway obstruction based on prenatal images. All 4 patients had an uncomplicated vaginal delivery. CONCLUSIONS: Technological advances in prenatal ultrasonography and magnetic resonance imaging have improved the ability to diagnose congenital abnormalities in utero. This allows for proper assessment of the airway to prevent any unexpected problems at delivery. We believe that many airway emergencies can be avoided by prenatal imaging and initiation of airway management in the prenatal period. PMID- 15897418 TI - Asymmetry of the vocal folds in patients with vocal fold immobility. AB - OBJECTIVES: To measure the vocal fold length (VFL) during inspiration and phonation and to determine the vertical difference of the vocal folds during phonation in patients with unilateral vocal fold immobility. DESIGN: Prospective study. SETTING: University hospital. Patients Thirty patients with unilateral vocal fold immobility. INTERVENTIONS: Each subject was asked to sustain the vowel /a/ and, after a short rest, to inhale slowly. The region over the larynx was scanned using multislice helical computed tomography during each maneuver; 3 dimensional endoscopic, coronal, and sagittal images were produced. MAIN OUTCOME MEASURES: The VFLs on each side and the vertical differences between the vocal folds were calculated. RESULTS: The inspiratory VFL on both sides was significantly longer in men than in women. It was significantly longer on the healthy side than on the immobile side in both groups. On the healthy side, the inspiratory VFL was significantly longer than the phonatory VFL in men, but there was no significant difference in women. In contrast, on the immobile side, the phonatory VFL was significantly longer than the inspiratory VFL in women, but there was no significant difference in men. The VFLs of the healthy and immobile sides varied in tandem. The immobile vocal fold was situated lower than the healthy vocal fold during phonation in 3 patients. CONCLUSIONS: Multislice helical computed tomography is a novel method to measure the VFL and the vertical level difference between the vocal folds. Application of this method might facilitate further understanding of laryngeal behavior in patients with unilateral vocal fold immobility. PMID- 15897419 TI - A study of TRAIL receptors in squamous cell carcinoma of the head and neck. AB - OBJECTIVE: To determine the potential immediate applicability of tumor necrosis factor-related apoptosis-inducing ligand receptor 1 (TRAIL-R1) and TRAIL-R2, the apoptotic forms of TRAIL-Rs, for preclinical testing. DESIGN AND SETTING: Head and neck squamous cell carcinoma (HNSCC) tumors were studied for TRAIL-R1 and TRAIL-R2 expression by immunohistochemical analysis. In addition, matched tumor and peripheral blood DNA samples were screened for 2 known TRAIL-R1 coding single nucleotide polymorphisms (C626G and G422A). Subjects Tumor samples taken from 43 patients (37 samples for immunohistochemical analysis and 6 additional ones included for polymorphism analysis). MAIN OUTCOME MEASURES: The expression of TRAIL-R1 and TRAIL-R2 and the presence of the TRAIL-R1 polymorphisms C626G and G422A. RESULTS: Fewer than 25% of HNSCC tumor cells expressed TRAIL-R1 and TRAIL R2. Surrounding tumor-infiltrating polymorphonuclear cells expressed TRAIL-R1 and TRAIL-R2 in 12 (32%) and 14 (38%) of cases, respectively. The TRAIL-R1 polymorphisms C626G and G422A were present in 36 (88%) and 33 (89%) cancer cases, respectively. Compared with control groups from another study, these polymorphism frequencies were statistically significant (P = .01 and .003, respectively). CONCLUSIONS: TRAIL-R expression was detected in less than half of the tumor specimens studied but not in any surrounding normal tissue and was found in a higher frequency on tumor-infiltrating polymorphonuclear cells than on tumor cells. These findings support the idea that the presence of TRAIL-Rs on some HNSCC tumors may make them more susceptible to apoptosis, and they also suggest that TRAIL-R-associated mechanisms may result in immune-modulatory effects on tumor-infiltrating polymorphonuclear cells. Furthermore, the significant association of somatic TRAIL-R1 genetic polymorphisms in this sample of patients with HNSCC suggests a potential association between constitutive TRAIL-R1 polymorphisms and development of HNSCC. Defining TRAIL-R expression and genetic polymorphisms in HNSCC represents the first step in examining TRAIL-related mechanisms for their potential as therapeutic targets. PMID- 15897420 TI - Usefulness of the LigaSure vessel sealing system during superficial lobectomy of the parotid gland. AB - OBJECTIVE: To evaluate the usefulness of the LigaSure Precise instrument in superficial lobectomy of the parotid gland. DESIGN: Prospective study of the surgical procedures in the LigaSure Vessel Sealing System and comparison with a conventionally treated control group. SETTING: Secondary care academic referral center. Patients Thirty-five patients with a parotid gland benign tumor were randomly allocated to 2 superficial lobe parotidectomy groups: 17 using the LigaSure procedure (group A) and 18 using the conventional method (group B). MAIN OUTCOME MEASURES: During the past few years, different methods of achieving hemostasis in parotid gland surgery have been tested as means of decreasing operative time and facial nerve injuries by controlling bleeding. With the whole LigaSure Vessel Sealing System, we experienced the usefulness of the LigaSure Precise instrument in superficial lobectomy of the parotid gland. RESULTS: No statistically significant differences were noted between the 2 groups in mean age, tumor diameter, length of hospital stay, time to return to work, or number of adverse events during or after surgery. Operative time was significantly shorter in group A than in group B (P < .001). Total operative time for conservative partial parotidectomy with traditional excision ranged from 115 to 235 minutes (mean, 155.8 minutes). Using the LigaSure system, the mean operative time was 136.4 minutes. Salivary fistulas were more common in group A (3 of 17 patients), with no cases in group B. CONCLUSIONS: The LigaSure method is comparable with but not superior to the conventional method. The main advantages of the LigaSure system are its "sutureless technique" and operative time savings; however, the cost is considerably higher. PMID- 15897421 TI - An in vivo evaluation of docetaxel delivered intratumorally in head and neck squamous cell carcinoma. AB - OBJECTIVE: To identify activity and biological mechanisms of intratumoral (IT) docetaxel on head and neck squamous cell carcinoma (HNSCC). METHODS: Docetaxel IT therapy was tested in xenograft models of 2 HNSCC lines, HN30 and HN12. The overall and disease-free survival rates, tumor growth, and toxic effects were measured. The pharmacokinetic profiles of docetaxel in plasma and tumor were compared after IT and intravenous (IV) administration. Comparisons between common and supradoses of docetaxel with regard to expression of regulators in the cell cycle, apoptosis, and signal transduction pathways were determined using Western blot analysis. RESULTS: In the HN30 and HN12 xenograft models, IT docetaxel improved overall as well as disease-free survival and reversed tumor growth. The only toxic effects noted were local (alopecia and skin breakdown). Skin breakdown resolved in all cases. At equivalent dosing levels, IT docetaxel achieved a 26 fold higher peak tumor concentration and 24-fold longer tumor exposure than IV treatment. Furthermore, limited plasma exposure was noted with IT docetaxel. Supradose levels of docetaxel produced distinct protein expression patterns for regulators of the cell cycle (cyclins A and B, p21, and p27), apoptosis (cleaved caspase-3 and cleaved PARP), and signal transduction (EGFR, pEGFR, pc-Jun, and pERK) in HNSCC, which supports a distinctive mechanism of action for supradose docetaxel levels. Since levels of cleaved caspase-3 and PARP, markers of apoptosis, were only elevated with lower doses, the observed cell death at supradose levels was probably due to necrosis. CONCLUSIONS: Injections of IT docetaxel at usual and supradoses are associated with a pharmacokinetic profile and biological mechanism distinct from those observed with usual IV doses. It is calculated that IT therapy in men will increase peak concentrations of docetaxel in tumors by 1000-fold over the conventional IV dose used clinically. These preclinical results support further testing of IT docetaxel in HNSCC. PMID- 15897422 TI - Extension of intracranial thrombosis after unilateral dissection of the internal jugular vein. AB - OBJECTIVE: Resection of the internal jugular vein can be necessary in cases of radical neck dissection or even in cases involving benign neoplasms such as glomus tumors. According to the triad described by Virchow (ie, stasis of blood flow, damage to the endothelium, and changes in hemostasis), the development of a thrombosis in the venous vessels superior to the resected internal jugular vein seems to be possible. The aim of this study was to determine the extension of possible intracranial thrombosis after wound healing. DESIGN: A total of 17 patients requiring resection of the internal jugular vein were evaluated prospectively using magnetic resonance imaging and 2-dimensional (time-of-flight) magnetic resonance angiography after the surgical treatment. Preoperative magnetic resonance images were obtained in all 17 patients for staging purposes as well as to exclude preoperative thrombosis of the internal jugular vein. Sixteen patients had malignant tumors, and 1 patient had a glomus tumor. RESULTS: Intracranial thrombosis with thrombosis of the sigmoid sinus was found in 4 patients, and thrombosis of the transverse sinus was found in 3 patients. There were no complications such as intracranial hemorrhage or signs of increased intracranial pressure in any patients. There was no evidence of intracranial thrombosis in 10 cases. In these cases, thrombosis of the venous vessels superior to the resected vein ranged from none to complete thrombosis of the superior bulb of the internal jugular vein. After unilateral resection of the internal jugular vein, the venous blood leaves the brain mainly via the venous network of the contralateral side of the neck. CONCLUSIONS: Although intracranial thrombosis of the sigmoid or transverse sinus seems to occur more frequently than was previously thought, intracranial complications such as venous infarction or increased intracranial pressure appear to be very rare. After unilateral radical neck dissection, the venous blood leaves the brain mainly via the venous system of the other side of the neck and the ipsilateral collateral veins. PMID- 15897423 TI - Comparison of ultrasonography and conventional radiography in the diagnosis of nasal fractures. AB - OBJECTIVE: To compare the diagnostic findings of ultrasonography and radiography in nasal fractures. DESIGN AND MAIN OUTCOME MEASURES: In this prospective study, 63 patients (23 female and 40 male; mean age, 26.8 years) with clinical signs of a nasal bone fracture were investigated. All patients underwent radiography (lateral view of the nose plus occipitomental view) and ultrasonography (10-MHz ultrasound scanner) of the nasal dorsum and the lateral nasal walls and a clinical examination by 2 consultants. Thirty-six patients underwent nasal fracture reduction. Two radiographs and 3 ultrasound images of each patient were analyzed by 2 experienced readers at different times. After assessing the nasal dorsum and lateral nasal walls in radiographs and ultrasound images, they decided whether the nose was fractured or not or whether the results were uncertain. The results were analyzed by various statistical testing methods (for sensitivity, specificity, positive and negative predictive value, and accuracy). RESULTS: Assessment of the lateral nasal walls revealed that ultrasonography was statistically superior (P = .04) to radiography. In contrast, assessment of the nasal dorsum showed radiography to be statistically superior (P = .01) to ultrasonography. Assessment of the nasal pyramid revealed no statistical difference between radiography and ultrasonography (P = .91). CONCLUSION: In assessment of the nasal pyramid, ultrasonography can be considered an alternative to radiography, with equivalent diagnostic performance. PMID- 15897424 TI - Outcomes of hyoid suspension for the treatment of obstructive sleep apnea. AB - OBJECTIVE: To examine the efficacy of hyoid myotomy and suspension as a treatment of hypopharyngeal obstruction in obstructive sleep apnea. DESIGN: Prospective, observational study. SETTING: Academic medical center. PATIENTS: Twenty-nine consecutive male patients with suspected hypopharyngeal obstruction INTERVENTIONS: Patients underwent hyoid suspension. Uvulopalatopharyngoplasty with or without tonsillectomy was performed at the same time for those patients who had not undergone this procedure previously. Patients underwent clinical examination and sleep study prior to surgery and approximately 1 year postoperatively. MAIN OUTCOME MEASURES: Primary outcome was a successful surgical result, defined as apnea-hypopnea index lower than 20, 50% or greater decline in apnea-hypopnea index, and no oxygen desaturations below 85% on the postoperative sleep study. Secondary outcomes included daytime sleepiness as determined by the Epworth Sleepiness Scale and the severity of snoring. Postoperative complications were also recorded. RESULTS: Only 5 (17%) of 29 patients achieved a successful outcome. The respiratory disturbance index did not change significantly for the group as a whole, although the lowest oxygen saturation did show some improvement. CONCLUSIONS: Hyoid suspension does not provide results equivalent to those reported for genioglossus advancement or multisession tongue radiofrequency. Hyoid suspension alone is not an efficacious treatment for hypopharyngeal airway obstruction in most patients with obstructive sleep apnea. PMID- 15897425 TI - Porous high-density polyethylene for orbital reconstruction. AB - OBJECTIVE: To determine the safety and efficacy of using porous high-density polyethylene (PHDPE) in the repair of orbital defects. DESIGN: Retrospective case series. SETTING: Academic tertiary care trauma center. Patients One hundred seventy patients with orbital defects requiring surgical repair. Intervention Orbital defect repair with PHDPE. Main Outcome Measure Our review documents surgical results and complications associated with the use of PHDPE. RESULTS: There was a 6.4% complication rate associated with the use of PHDPE. The infection rate was 1.8%. The persistent orbital malposition rate was 3.5%. The extrusion rate was 0%. CONCLUSIONS: This report represents the largest case series in the literature using PHDPE for orbital reconstructions. The use of PHDPE resulted in a low complication rate and excellent functional and cosmetic reconstructive results. Because of our success with the use of PHDPE, we have changed our clinical practice to minimize the use of autologous graft material, thereby eliminating donor site morbidity in cases involving orbital reconstruction. PMID- 15897426 TI - Congenital cricopharyngeal achalasia: management with botulinum toxin before myotomy. PMID- 15897427 TI - Pseudoaneurysm of the internal carotid artery: a case report and review of the literature. PMID- 15897428 TI - Meniere's disease or migraine? The clinical significance of fluctuating hearing loss with vertigo. PMID- 15897429 TI - Meniere's Disease and Migraine. PMID- 15897430 TI - Radiology quiz case 1. Primary solitary hemangioma of the thyroid gland. PMID- 15897431 TI - Radiology quiz case 2. Reparative granuloma. PMID- 15897434 TI - Pathology quiz case. Pleomorphic adenoma. PMID- 15897436 TI - George Washington University School of Medicine and Department of Surgery. PMID- 15897437 TI - To impart this art: the didactic mission. PMID- 15897438 TI - A 12-year prospective study of postinjury multiple organ failure: has anything changed? AB - HYPOTHESIS: The incidence and severity of postinjury multiple organ failure (MOF) has decreased over the last decade. DESIGN: A prospective 12-year inception cohort study ending December 31, 2003. SETTING: Regional academic level I trauma center. PATIENTS: One thousand three hundred forty-four trauma patients at risk for postinjury MOF. Inclusion criteria were aged older than 15 years, admission to the trauma intensive care unit, an Injury Severity Score higher than 15, and survival for more than 48 hours after injury. Isolated head injuries were excluded from this study. Previously identified risk factors for postinjury MOF were age, Injury Severity Score, and receiving a blood transfusion within 12 hours of injury. MAIN OUTCOME MEASURES: Multiple organ failure was defined by a Denver MOF score of 4 or more for longer than 48 hours after injury. Multiple organ failure severity was defined by the maximum daily MOF score and the number of MOF free days within the first 28 postinjury days. RESULTS: Multiple organ failure was diagnosed in 339 (25%) of 1244 patients. The mean age and Injury Severity Scores increased and the use of blood transfusion during resuscitation decreased over the 12-year study period. After adjusting for age, injury severity, and amount of blood transfused during resuscitation, there was a decreased incidence of MOF over the study period. Of the patients who developed MOF, there was a decrease in disease severity and duration as measured by the maximum daily MOF score and the MOF free days. Although the overall mortality rate remained constant, the MOF-specific mortality decreased. CONCLUSIONS: The incidence, severity, and attendant mortality of postinjury MOF decreased over the last 12 years despite an increased MOF risk. Improvements in MOF outcomes can be attributed to improvements in trauma and critical care and are associated with decreased use of blood transfusion during resuscitation. PMID- 15897439 TI - Spectrum of esophageal motility disorders: implications for diagnosis and treatment. AB - BACKGROUND: The named primary esophageal motility disorders (PEMDs) are achalasia, diffuse esophageal spasm (DES), nutcracker esophagus (NE), and hypertensive lower esophageal sphincter (HTN-LES). Although the diagnosis and treatment of achalasia are well defined, such is not the case with the other disorders. HYPOTHESIS: (1) Symptoms do not reliably distinguish PEMDs from gastroesophageal reflux disease; (2) esophageal function tests are essential to this distinction and to identifying the type of PEMD; (3) minimally invasive surgery is effective for each condition; and (4) the laparoscopic approach is better than the thoracoscopic approach. DESIGN: University hospital tertiary care center. SETTING: Retrospective review of a prospectively collected database. PATIENTS AND METHODS: A diagnosis of PEMD was established in 397 patients by esophagogram, endoscopy, manometry, and pH monitoring. There were 305 patients (77%) with achalasia, 49 patients (12%) with DES, 41 patients (10%) with NE, and 2 patients (1%) with HTN-LES. Two hundred eight patients (52%) underwent a myotomy by either a thoracoscopic or a laparoscopic approach. RESULTS: Ninety nine patients (25%) had a diagnosis of gastroesophageal reflux disease at the time of referral and had been treated with acid-suppressing medications. In achalasia and DES, a thoracoscopic or laparoscopic myotomy relieved dysphagia and chest pain in more than 80% of the patients. In contrast, in NE the results were less predictable, and the operation most often failed to relieve symptoms. CONCLUSIONS: These results show that (1) symptoms were unreliable in distinguishing gastroesophageal reflux disease from PEMDs; (2) esophageal function tests were essential to diagnose PEMD and to define its type; (3) the laparoscopic approach was better than the thoracoscopic approach; (4) a laparoscopic Heller myotomy is the treatment of choice for achalasia, DES, and HTN-LES; and (5) a predictably good treatment for NE is still elusive, and the results of surgery were disappointing. PMID- 15897440 TI - Critical appraisal of the clinical and pathologic predictors of survival after resection of large hepatocellular carcinoma. AB - HYPOTHESIS: A subset of patients with hepatocellular carcinoma (HCC) with a diameter of 10 cm or larger may benefit from hepatic resection. DESIGN: Retrospective study of a multi-institutional database. SETTING: Five major hepatobiliary centers. PATIENTS: We identified 300 patients who underwent hepatic resection for HCC 10 cm or larger. MAIN OUTCOME MEASURES: Clinical and pathologic data were collected, and prognostic factors were evaluated by univariate and multivariate analyses. Patient survival was stratified according to a clinical scoring system and pathologic T classification. RESULTS: The perioperative mortality rate was 5%. At a median follow-up of 32 months, the median survival was 20.3 months, and the 5-year actuarial survival rate was 27%. Four clinical factors-alpha-fetoprotein of 1000 ng/mL or higher, multiple tumor nodules, the presence of major vascular invasion, and the presence of severe fibrosis-were significant predictors of poor survival (all P<.05). Patients were assigned a clinical score according to the following risk factors: 1, no factor; 2, one or two factors; or 3, three or four factors. On the basis of the clinical score, patients could be stratified into only 2 distinct prognostic groups: no factor (score of 1) vs 1 or more factors (score of 2 or 3) (P<.001). In contrast, when patients were stratified according to pathologic T classification, 3 distinct groups were identified: T1 vs T2 vs T3 and T4 combined (P<.001). Fifty-six percent of the patients with a clinical score of 2 and 20% of patients with a clinical score of 3 actually had T1 or T2 disease on pathologic examination. CONCLUSIONS: Patients with large HCCs should be considered for liver resection as this treatment is associated with a 5-year survival rate exceeding 25%. Clinical predictors should not be used to exclude patients from surgical resection because these factors do not reliably predict outcome. PMID- 15897441 TI - Liver transplantation for hepatocellular carcinoma: expanding special priority to include stage III disease. AB - HYPOTHESIS: After liver transplantation, patients with stage III hepatocellular carcinoma (HCC) experience survivals similar to those of patients with less advanced disease and of matched control subjects. DESIGN: Retrospective review of prospectively collected database. SETTING: University hospital. PATIENTS: Fifty one adults with HCC and 153 matched adults without HCC who underwent orthotopic liver transplantation. MAIN OUTCOME MEASURES: One-, 3-, and 5-year survivals for all groups. After matching for year of transplantation, age, sex, and underlying liver disease, long-term survival was compared between groups. Rates of recurrence were also measured in the HCC groups. RESULTS: From August 1, 1985, to February 28, 2002, we performed 635 adult liver transplantations, including 51 (8%) in patients with HCC. One hundred fifty-one patients without HCC who underwent transplantation were selected as controls. Patient demographic features were similar between case-control groups. The overall 5-year survival trend was worse for patients with HCC vs their matched controls (48% vs 65%; P = .07); however, this survival disadvantage was eliminated when patients with stages I through III HCC were combined and compared with their matched controls (59% vs 63%; P = .96). Survival of patients with stage III disease was comparable to that of matched controls (65% vs 59%; P = .44). CONCLUSIONS: For patients with stages I through III disease, long-term survival is comparable to that of matched controls, and only patients with stage IV disease experience poorer survival. Consideration should be given to granting exception points to patients with stage III disease. PMID- 15897442 TI - Survival after pediatric liver transplantation: why does living donation offer an advantage? AB - HYPOTHESIS: Living donor liver transplantation (LDLT) results in improved survival compared with deceased whole and split organ transplantation in children. OBJECTIVE: To evaluate the effect of LDLT on graft and patient survival in pediatric liver transplantation. DESIGN: Retrospective cohort. METHODS: Data included all pediatric recipients (aged <18 years) registered in the UNOS (United Network for Organ Sharing) database from October 1, 1987, to May 24, 2004. Covariates predictive of survival by univariate analyses were included in the Cox proportional hazards regression models in a blockwise fashion to determine predictors of survival. RESULTS: Kaplan-Meier graft and patient survival rates were improved in LDLT recipients compared with recipients of deceased whole and split organ transplantations (P<.01). In the initial model (model P<.001), prognostic factors for graft and patient survival included recipient age, race, origin of liver disease, certain pretransplantation laboratory data, medical condition, multiorgan transplantation, retransplantation, recipient-donor ABO blood compatibility, and cold and warm ischemia times. The addition of graft type to the initial covariate set did not significantly change the model (P = .21, covariate P = .09). However, most of the positive prognostic factors identified in the model were inherent characteristics of LDLT recipients and the LDLT procedure. CONCLUSIONS: Graft and patient survival in the pediatric population is better with LDLT than deceased organ transplantation. Factors that contribute to this difference include recipients who are less ill, who have shorter cold and warm ischemia times, and those with a decreased need for retransplantation but not the type of graft per se. PMID- 15897443 TI - Primary hyperparathyroidism surgical management since the introduction of minimally invasive parathyroidectomy: Mayo Clinic experience. AB - HYPOTHESIS: Minimally invasive parathyroidectomy (MIP) for primary hyperparathyroidism (HPT) has equal cure and recurrence rates as standard cervical exploration. Changes in the management of primary HPT have occurred since introducing MIP including localization, anesthesia, intraoperative parathyroid hormone monitoring, and indications for parathyroidectomy. DESIGN: Cohort analysis of 1361 consecutive patients with primary HPT operated on at the Mayo Clinic, Rochester, Minn, from June 1998 through March 2004. Mean follow-up, 25 months. SETTING: Tertiary referral center. PATIENTS: One thousand three hundred sixty-one patients operated on for primary HPT, excluding 160 patients who were reoperated on. INTERVENTION: Standard cervical exploration MIP. MAIN OUTCOME MEASURES: Cure, recurrence, localization, anesthesia, hospitalization, intraoperative parathyroid hormone level monitoring, contraindications to MIP, surgical indications, assessment of osteoporosis and osteopenia, postoperative patient assessment of general patient health, and operative satisfaction. RESULTS: Cure of primary HPT for both conventional exploration and MIP was 97%; only 1 patient who underwent MIP had a potential recurrence. Imaging sensitivity and positive predictive values were as follows: sestamibi scintigraphy, 86% and 93%; ultrasonography, 61% and 87%, respectively. Usage of general vs local anesthesia with intravenous sedation was 46% and 49%, respectively, in patients w ho underwent MIP; 46% were dismissed as outpatients, 49% had single-night stays. The accuracy of intraoperative parathyroid hormone level monitoring was as follows: 98% (8% had true-negative results); the frequency of multiple gland disease was 13%. Accounting for causes precluding MIP, an estimated 60% to 70% of all patients would be eligible for MIP. By preoperative assessment, 79% had osteoporosis-osteopenia; 58% with postoperative bone mineral density measurements were improved. More than 85% were satisfied with the results of their operation. CONCLUSION: With high-quality localization and intraoperative parathyroid hormone level monitoring, MIP can be performed with equal cure rates as standard cervical exploration, with no present evidence of delayed recurrence. PMID- 15897444 TI - Carotid artery stents for blunt cerebrovascular injury: risks exceed benefits. AB - BACKGROUND: Carotid stenting has been advocated in patients with grade III blunt carotid artery injuries (hereafter referred to as "blunt CAIs") because of the persistence of the pseudoaneurysm and concern for subsequent embolization or rupture. HYPOTHESIS: Carotid stenting is safe and effective for blunt CAIs. DESIGN: Analysis of a prospective database of all patients with blunt CAIs. SETTING: A state-designated, level I, urban trauma center. PATIENTS AND METHODS: In January 1, 1996, we initiated comprehensive screening for blunt CAIs with angiography based on injury patterns. Patients without contraindications receive anticoagulation therapy immediately for documented lesions. Patients with persistent pseudoaneurysms on a second angiography at 7 to 10 days after injury are candidates for stent placement. RESULTS: During the study period (January 1, 1996, to May 1, 2004), 46 patients sustained blunt carotid pseudoaneurysms; 23 (50%) underwent carotid stent placement. There were 4 complications in patients undergoing carotid stent placement: 3 strokes and 1 subclavian dissection. Follow up angiography was performed in 38 patients (18 patients with stents who received antithrombotic agents, 20 patients who received antithrombotic agents alone); 8 patients had poststent carotid occlusion despite having received concurrent anticoagulation therapy. Carotid occlusion rates were significantly different (45% in patients with stents vs 5% in those who received antithrombotic agents alone). In the patients not undergoing stent placement, the only complication was a middle cerebral artery stroke in a patient not treated with antithrombotic therapy. CONCLUSIONS: Patients who have carotid stents placed for blunt carotid pseudoaneurysms have a 21% complication rate and a documented occlusion rate of 45%. In contrast, patients treated with antithrombotic agents alone had an occlusion rate of 5%; no asymptomatic patient treated with antithrombotic agents for their injury had a stroke. Antithrombotic therapy remains the recommended therapy for blunt CAIs, but the role of intraluminal stents remains to be defined. PMID- 15897445 TI - Infragenicular polytetrafluoroethylene bypass with distal vein cuffs for limb salvage: a contemporary series. AB - HYPOTHESIS: Infragenicular polytetrafluoroethylene (PTFE)-venous cuff bypass grafting provides acceptable graft patency and limb salvage rates for limb salvage. DESIGN: Retrospective clinical review of a consecutive series. SETTING: Vascular surgical practice during the interval October 1, 2000, to September 1, 2004. PATIENTS: Fifty-one male and 49 female patients whose mean age was 76.9 years were operated on for tissue loss (67%), chronic rest pain (28%), and severe claudication (6%). Fifty-two percent of patients were diabetic and 49% had undergone previous leg bypass surgery. All patients had absent or inadequate greater saphenous vein, and 84 patients had absent or inadequate arm vein. INTERVENTIONS: One hundred five infragenicular PTFE bypasses were performed in these 100 patients. Distal targets were the infragenicular popliteal (40), posterior tibial (35), anterior tibial (16), and peroneal arteries (14). Sixty eight venous cuffs were constructed from lesser saphenous vein. MAIN OUTCOME MEASURES: Graft patency, limb salvage, and patient survival were analyzed. RESULTS: Twelve early graft failures resulted in 7 leg amputations. The mean +/- SE 3-year primary patency and limb salvage rates were 64.4% +/- 12.8% and 74.4% +/- 11.9%, respectively. Perioperative mortality was 2.9% and 3-year survival was 38%. Graft follow-up ranged from 1 to 47 months with a mean of 13 months using life-table methods. CONCLUSIONS: For patients requiring arterial revascularization for limb salvage, in which autologous venous conduit is unavailable, distal venous cuff-PTFE bypass provides acceptable patency and limb salvage rates when viewed in the context of short life expectancy for these elderly patients. PMID- 15897446 TI - Are young surgeons competent to perform alimentary tract surgery? AB - BACKGROUND: Assessment of competency during residency training has received increased attention recently. There has been less attention given to the competency of residents after training. HYPOTHESIS: Patient outcomes for alimentary tract surgery (ATS) should be similar for surgeons who recently completed their residency training compared with more experienced surgeons, indicating that the younger surgeons had achieved clinical competency on completion of their residency training. DESIGN: Retrospective analysis of Illinois inpatient discharge data (January 1, 1996-December 31, 1999). SETTING: All 205 nonfederal acute care hospitals in Illinois. PATIENTS: The patients were 120 160 adult Illinois residents who underwent ATS in Illinois. MAIN OUTCOME MEASURES: Mortality rate, morbidity rate, and hospital length of stay. RESULTS: Regression analyses demonstrated that surgeon experience was a significant determinant of mortality and morbidity rates, with worse outcomes observed for patients of young surgeons undergoing high-complexity ATS (ie, procedures other than appendectomy and cholecystectomy). CONCLUSIONS: For high-complexity ATS, there was a significant disparity in outcomes between young and more experienced surgeons, whereas for low-complexity ATS, there was no disparity. Attention to competency during residency training is warranted, especially as it relates to high-complexity ATS. Furthermore, patient outcomes provide an opportunity to assess competency after training that can complement assessments during training and together identify educational strengths and weaknesses of residency training. PMID- 15897447 TI - Surgical sphincteroplasty in 446 patients. AB - HYPOTHESIS: Pancreaticobiliary sphincter disease is reliably diagnosed by endoscopic and intraoperative manometry. DESIGN: Retrospective review of prospectively collected data. SETTING: A 400-bed urban university hospital. PATIENTS: Between May 1, 1978, and March 27, 2002, 446 patients were treated surgically for dysfunction of the pancreaticobiliary sphincters. There were 376 females and 70 males (mean +/- SD age, 41.6 +/- 12.5 years). There were 372 patients with sphincter of Oddi dysfunction, and 74 with pancreas divisum. Symptoms included abdominal pain (100.0%), nausea/vomiting (80.5%), back pain (57.2%), and pancreatitis (22.4%). INTERVENTIONS: Perfusion manometry has evolved as the gold standard for diagnosis, and intraoperative manometry was done in 214 patients. All patients underwent transduodenal sphincteroplasty and biopsies of the ampullae and transampullar septa. RESULTS: Excellent or good results were seen in 86.8% of the patients with sphincter of Oddi dysfunction and in 63.5% of the patients with pancreas divisum. Common duct and sphincter of Oddi pressures were 0 mm Hg in all patients after sphincteroplasty. Pancreatic duct and pancreatic sphincter manometry results were improved in 82.4% of the patients. Biopsy results of the main and accessory sphincters demonstrated inflammation and/or fibrosis in 33.9% of ampullae and 43.5% of transampullar septa, but this did not correlate with outcome. There was 1 death from a duodenal leak. Complications occurred in 34.8% of patients, with pancreatitis (8.8%), asymptomatic hyperamylasemia (6.0%), and wound/abdominal infection (7.1%) the most common. Predictive factors for good outcome were reduction in pancreatic duct and sphincter pressures following sphincteroplasty. CONCLUSION: Good to excellent results may be achieved by surgical sphincteroplasty when careful patient selection by manometry is used. PMID- 15897448 TI - Image of the month. Heterotopic pancreas. PMID- 15897449 TI - Targeting quantum dots to surface proteins in living cells with biotin ligase. AB - Escherichia coli biotin ligase site-specifically biotinylates a lysine side chain within a 15-amino acid acceptor peptide (AP) sequence. We show that mammalian cell surface proteins tagged with AP can be biotinylated by biotin ligase added to the medium, while endogenous proteins remain unmodified. The biotin group then serves as a handle for targeting streptavidin-conjugated quantum dots (QDs). This labeling method helps to address the two major deficiencies of antibody-based labeling, which is currently the most common method for targeting QDs to cells: the size of the QD conjugate after antibody attachment and the instability of many antibody-antigen interactions. To demonstrate the versatility of our method, we targeted QDs to cell surface cyan fluorescent protein and epidermal growth factor receptor in HeLa cells and to alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptors in neurons. Labeling requires only 2 min, is extremely specific for the AP-tagged protein, and is highly sensitive. We performed time-lapse imaging of single QDs bound to AMPA receptors in neurons, and we compared the trafficking of different AMPA receptor subunits by using two color pulse-chase labeling. PMID- 15897450 TI - Overexpression of LMO4 induces mammary hyperplasia, promotes cell invasion, and is a predictor of poor outcome in breast cancer. AB - The zinc finger protein LMO4 is overexpressed in a high proportion of breast carcinomas. Here, we report that overexpression of a mouse mammary tumor virus (MMTV)-Lmo4 transgene in the mouse mammary gland elicits hyperplasia and mammary intraepithelial neoplasia or adenosquamous carcinoma in two transgenic strains with a tumor latency of 13-18 months. To investigate cellular processes controlled by LMO4 and those that may be deregulated during oncogenesis, we used RNA interference. Down-regulation of LMO4 expression reduced proliferation of human breast cancer cells and increased differentiation of mouse mammary epithelial cells. Furthermore, small-interfering-RNA-transfected breast cancer cells (MDA-MB-231) had a reduced capacity to migrate and invade an extracellular matrix. Conversely, overexpression of LMO4 in noninvasive, immortalized human MCF10A cells promoted cell motility and invasion. Significantly, in a cohort of 159 primary breast cancers, high nuclear levels of LMO4 were an independent predictor of death from breast cancer. Together, these findings suggest that deregulation of LMO4 in breast epithelium contributes directly to breast neoplasia by altering the rate of cellular proliferation and promoting cell invasion. PMID- 15897451 TI - VEGF-integrin interplay controls tumor growth and vascularization. AB - Cross-talk between the major angiogenic growth factor, VEGF, and integrin cell adhesion receptors has emerged recently as a critical factor in the regulation of angiogenesis and tumor development. However, the molecular mechanisms and consequences of this intercommunication remain unclear. Here, we define a mechanism whereby integrin alpha v beta3, through activation, clustering, and signaling by means of p66 Shc (Src homology 2 domain containing), regulates the production of VEGF in tumor cells expressing this integrin. Tumors with "activatable" but not "inactive" beta3 integrin secrete high levels of VEGF, which in turn promotes extensive neovascularization and augments tumor growth in vivo. This stimulation of VEGF expression depends upon the ability of alpha v beta3 integrin to cluster and promote phosphorylation of p66 Shc. These observations identify a link between beta3 integrins and VEGF in tumor growth and angiogenesis and, therefore, may influence anti-integrin as well as anti-VEGF therapeutic strategies. PMID- 15897452 TI - The candidate tumor suppressor ING4 represses activation of the hypoxia inducible factor (HIF). AB - The hypoxia inducible factor (HIF) plays an important role in the progression of a number of pathophysiological processes including tumorigenesis. In addition to several well characterized oxygen-dependent modes of regulation, the function of the HIF transcription factor can also be influenced through the action of other regulatory pathways. Misregulation of these factors resulting in inappropriate HIF expression or activity can contribute to the progression of human cancers through the induction of genes promoting angiogenesis, glycolysis, cell survival, and metastasis, among other processes. The candidate tumor suppressor protein inhibitor of growth family member 4 (ING4) has recently been implicated as a repressor of angiogenesis and tumor growth through association with NF-kappaB. Here we demonstrate that suppression of ING4 further induces HIF transcriptional activity as well. ING4 directly associates with the HIF prolyl hydroxylase, an Fe(II)-dependent oxygenase previously shown to mediate HIF stability as a function of oxygen availability. However, rather than affecting HIF's stability, ING4 mediates HIF's activity. These data support a model in which, in addition to regulating HIF stability, HIF prolyl hydroxylases can modulate HIF function through the recruitment of ING4, a likely component of a chromatin-remodeling complex. PMID- 15897453 TI - Components of the REST/CoREST/histone deacetylase repressor complex are disrupted, modified, and translocated in HSV-1-infected cells. AB - The infected cell protein (ICP)0 enables gene expression and the replication of herpes simplex virus (HSV)-1 in cells infected at low multiplicities and enhances the expression of genes introduced into cells by transfection or infection. We report that a short sequence of ICP0 is similar to a sequence in the amino terminus of CoREST, a corepressor that exists in complexes with the repressor REST and histone deacetylases (HDACs) 1 or 2 to repress cellular gene expression. In wild-type-virus-infected cells, HDAC1 dissociates from the CoREST/REST complex, CoREST and HDAC1 are phosphorylated by a process mediated by viral protein kinases, and CoREST and HDAC1 are partially translocated to the cytoplasm. In cells infected with a virus mutant (DeltaICP4), in which ICP0 accumulates, but post-alpha gene expression is blocked, HDAC1 is dissociated from the CoREST/REST complex, but translocation to the cytoplasm does not occur. After infection with a mutant virus from which ICP0 is deleted, the complex remains intact, but, under conditions of productive infection, the complex is partially translocated to the cytoplasm. These results suggest that, at low multiplicities of infection, ICP0 blocks CoREST-mediated silencing of viral genes by dissociation of HDAC1, whereas subsequent modifications and translocation of the components of the complex are the functions of other viral gene products made later in infection. PMID- 15897454 TI - A prokaryotic superoxide dismutase paralog lacking two Cu ligands: from largely unstructured in solution to ordered in the crystal. AB - Little is known about prokaryotic homologs of Cu,Zn superoxide dismutase (SOD), an enzyme highly conserved among eukaryotic species. In 138 Archaea and Bacteria genomes, 57 of these putative homologs were found, 11 of which lack at least one of the metal ligands. Both the solution and the crystal structures of the SOD like protein from Bacillus subtilis, lacking two Cu ligands and found to be enzymatically inactive, were determined. In solution, the protein is monomeric. The available nuclear Overhauser effects, together with chemical-shift index values, allowed us to define and to recognize the typical Cu,Zn SOD Greek beta barrel but with largely unstructured loops (which, therefore, sample a wide range of conformations). On the contrary, in the crystal structure (obtained in the presence of slight excess of Zn), the protein is well structured and organized in covalent dimers held by a symmetric bridge consisting of a Zn ion bound to an Asp His dyad in a tetrahedral geometry. Couples of dimers held by hydrophobic interactions and H bonds are further organized in long chains. The order/disorder transition is discussed in terms of metal binding and physical state. PMID- 15897455 TI - Quantitating intracellular transport of polyplexes by spatio-temporal image correlation spectroscopy. AB - Quantitatively understanding how nonviral gene delivery vectors (polyplexes) are transported inside cells is essential before they can be optimized for gene therapy and medical applications. In this study, we used spatio-temporal image correlation spectroscopy (ICS) to follow polymer-nucleic acid particles (polyplexes) of various sizes and analyze their diffusive-like and flow behaviors intracellularly to elucidate the mechanisms responsible for their transport. ICS is a quantitative imaging technique that allows the assessment of particle motion in complex systems, although it has not been widely used to date. We find that the internalized polyplexes are able to use microtubule motors for intracellular trafficking and exhibit different transport behaviors for short (<10 s) versus long (approximately 60 s) correlation times. This motion can be explained by a memory effect of the microtubule motors. These results reveal that, although microtubule motor biases may be present for short periods of time, resulting in a net directional velocity, the overall long-term motion of the polyplexes is best described as a random walk-like process. These studies suggest that spatio temporal ICS is a powerful technique for assessing the nature of intracellular motion and provides a quantitative tool to compare the transport of different objects within a living cell. PMID- 15897456 TI - A CACNA1F mutation identified in an X-linked retinal disorder shifts the voltage dependence of Cav1.4 channel activation. AB - Light stimuli produce graded hyperpolarizations of the photoreceptor plasma membrane and an associated decrease in a voltagegated calcium channel conductance that mediates release of glutamate neurotransmitter. The Ca(v)1.4 channel is thought to be involved in this process. The CACNA1F gene encodes the poreforming subunit of the Ca(v)1.4 channel and various mutations in CACNA1F cause X-linked incomplete congenital stationary night blindness (CSNB2). The molecular mechanism of the pathology underlying the CSNB2 phenotype remains to be established. Recent clinical investigations of a New Zealand family found a severe visual disorder that has some clinical similarities to, but is clearly distinct from, CSNB2. Here, we report investigations into the molecular mechanism of the pathology of this condition. Molecular genetic analyses identified a previously undescribed nucleotide substitution in CACNA1F that is predicted to encode an isoleucine to threonine substitution at CACNA1F residue 745. The I745T CACNA1F allele produced a remarkable approximately -30-mV shift in the voltage dependence of Ca(v)1.4 channel activation and significantly slower inactivation kinetics in an expression system. These findings imply that substitution of this wild-type residue in transmembrane segment IIS6 may have decreased the energy required to open the channel. Collectively, these findings suggest that a gain-of-function mechanism involving increased Ca(v)1.4 channel activity is likely to cause the unusual phenotype. PMID- 15897457 TI - Impact of an ectoparasite on the immunity and pathology of an invertebrate: evidence for host immunosuppression and viral amplification. AB - Varroa mites (Varroa destructor) are ectoparasites of honey bees (Apis mellifera) and cause serious damage to bee colonies. The mechanism of how varroa mites kill honey bees remains unclear. We have addressed the effects of the mites on bee immunity and the replication of a picorna-like virus, the deformed wing virus (DWV). The expression of genes encoding three antimicrobial peptides (abaecin, defensin, and hymenoptaecin) and four immunity-related enzymes (phenol oxidase, glucose dehydrogenase, glucose oxidase, and lysozyme) were used as markers to measure the difference in the immune response. We have demonstrated an example of an ectoparasite immunosuppressing its invertebrate host with the evidence that parasitization significantly suppressed expression of these immunity-related genes. Given that ticks immunosuppress their vertebrate hosts, our finding indicates that immunosuppression of hosts may be a common phenomenon in the interaction and coevolution between ectoparasites and their vertebrate and invertebrate hosts. DWV viral titers were significantly negatively correlated with the expression levels of the immunity-related enzymes. All bees had detectable DWV. Mite-infested pupae developed into adults with either normal or deformed wings. All of the deformed-wing bees were greatly infected by DWV (approximately 10(6) times higher than varroa-infested but normal-winged bees). Injection with heat-killed bacteria dramatically promoted DWV titers (10(5) times in 10 h) in the mite-infested, normal-winged bees to levels similar to those found in mite-infested, deformed-wing bees. Varroa mites may cause the serious demise of honey bees by suppressing bee immunity and by boosting the amplification of DWV in bees exposed to microbes. PMID- 15897458 TI - Peptide signaling during terminal differentiation of Dictyostelium. AB - A wide variety of mechanisms have evolved for intercellular communication in metazoans, but some of the signaling molecules were already used in their predecessors. The social amoeba, Dictyostelium discoideum, is known to use peptides to trigger sporulation within fruiting bodies, but their sequences have not been defined. We found that the peptide signal spore differentiation factor 2 (SDF-2) is processed from acyl-CoA binding protein, AcbA. The mammalian homolog of AcbA is processed to diazepam binding inhibitor that binds to the GABA(A) receptor in the brain and to peripheral 1,4 benzodiazepine receptors. Although Dictyostelium has neither GABA(A) nor peripheral-type benzodiazepine receptors, we find that both a diazepam binding inhibitor peptide and diazepam (Valium) can mimic SDF-2 in a Dictyostelium bioassay. Mutants lacking AcbA sporulate well only when developed in chimeras with WT cells. Using a yeast system we show that ligand binding to the SDF-2 receptor histidine kinase, DhkA, inhibits phosphorelay, which can account for its ability to induce rapid sporulation. PMID- 15897459 TI - Metal ion-binding properties of (N3)-deprotonated uridine, thymidine, and related pyrimidine nucleosides in aqueous solution. AB - The acidity constants for (N3)H of the uridine-type ligands (U) 5-fluorouridine, 5-chloro-2'-deoxyuridine, uridine, and thymidine (2'-deoxy-5-methyluridine) and the stability constants of the M(U-H)(+) complexes for M(2+) = Mg(2+), Ca(2+), Sr(2+), Ba(2+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Zn(2+), Cd(2+), and Pb(2+) were measured (potentiometric pH titrations; aqueous solution; 25 degrees C; I = 0.1 M, NaNO(3)). Plots of logK(M(U-H))(M) vs. pK(U)(H) result in straight lines that are compared with previous plots for simple pyridine-type and o amino(methyl)pyridine-type ligands as well as with the stabilities of the corresponding M(cytidine)(2+) complexes. The results indicate monodentate coordination to (N3)(-) in M(U-H)(+) for Co(2+) and Ni(2+). For the M(U-H)(+) species of Cd(2+), Zn(2+), or Cu(2+), increased stabilities imply that semichelates form, i.e., M(2+) is (N3)(-)-bound and coordinated water molecules form hydrogen bonds to (C2)O and (C4)O; these "double" semichelates are in equilibrium with "single" semichelates involving either (C2)O or (C4)O and possibly also with four-membered chelates for which M(2+) is innersphere coordinated to (N3)(-) and a carbonyl oxygen. For the alkaline earth ions, semichelates dominate with the M(2+) outersphere bound to (N3)(-) and innersphere to one of the carbonyl oxygens. Mn(U-H)(+) is with its properties between those of Cd(2+) (which probably also hold for Pb(2+)) and the alkaline earth ions. In nucleic acids, M(2+)-C(O) interactions are expected, if support is provided by other primary binding sites. (N3)H may possibly be acidified via carbonyl coordinated M(2+) to become a proton donor in the physiological pH range, at which direct (N3)(-) binding of M(2+) also seems possible. PMID- 15897460 TI - The crystal structures of human steroidogenic factor-1 and liver receptor homologue-1. AB - Steroidogenic factor-1 (SF-1) and liver receptor homologue-1 (LRH-1) belong to the fushi tarazu factor 1 subfamily of nuclear receptors. SF-1 is an essential factor for sex determination during development and regulates adrenal and gonadal steroidogenesis in the adult, whereas LRH-1 is a critical factor for development of endodermal tissues and regulates cholesterol and bile acid homeostasis. Regulatory ligands are unknown for SF-1 and LRH-1. A reported mouse LRH-1 structure revealed an empty pocket in a region commonly occupied by ligands in the structures of other nuclear receptors, and pocket-filling mutations did not alter the constitutive activity observed. Here we report the crystal structures of the putative ligand-binding domains of human SF-1 at 2.1-A resolution and human LRH-1 at 2.5-A resolution. Both structures bind a coactivator-derived peptide at the canonical activation-function surface, thus adopting the transcriptionally activating conformation. In human LRH-1, coactivator peptide binding also occurs to a second site. We discovered in both structures a phospholipid molecule bound in a pocket of the putative ligand-binding domain. MS analysis of the protein samples used for crystallization indicated that the two proteins associate with a range of phospholipids. Mutations of the pocket-lining residues reduced the transcriptional activities of SF-1 and LRH-1 in mammalian cell transfection assays without affecting their expression levels. These results suggest that human SF-1 and LRH-1 may be ligand-binding receptors, although it remains to be seen if phospholipids or possibly other molecules regulate SF-1 or LRH-1 under physiological conditions. PMID- 15897461 TI - The two-component signal transduction system RR06/HK06 regulates expression of cbpA in Streptococcus pneumoniae. AB - Streptococcus pneumoniae encounters a number of environmental niches in the body, including the nasopharynx, lungs, blood, middle ear, and brain. Recent studies have identified 13 putative two-component signal-transduction systems in S. pneumoniae, which are likely to be important for gene regulation in response to external stimuli. Here, we present conclusive evidence for the regulation of choline binding protein A (CbpA), a major pneumococcal virulence factor and protective antigen, by one of these two-component signal-transduction systems. We have demonstrated divergent expression of cbpA in unmarked hk06 and rr06 deletion mutants relative to wild-type S. pneumoniae D39 by using Western immunoblotting and real-time RT-PCR. Electrophoretic mobility-shift and solid-phase binding assays have demonstrated the binding of RR06 to the promoter region of cbpA, suggesting that RR06/HK06 directly regulates cbpA transcription. We have also shown that this system is important for the ability of the pneumococcus to adhere to epithelial cells in vitro and to survive and proliferate in an in vivo mouse model. Thus, the RR06/HK06 system has a significant role in pathogenesis, both in colonization and invasive disease. PMID- 15897462 TI - Regulatory on/off minimization of metabolic flux changes after genetic perturbations. AB - Predicting the metabolic state of an organism after a gene knockout is a challenging task, because the regulatory system governs a series of transient metabolic changes that converge to a steady-state condition. Regulatory on/off minimization (ROOM) is a constraint-based algorithm for predicting the metabolic steady state after gene knockouts. It aims to minimize the number of significant flux changes (hence on/off) with respect to the wild type. ROOM is shown to accurately predict steady-state metabolic fluxes that maintain flux linearity, in agreement with experimental flux measurements, and to correctly identify short alternative pathways used for rerouting metabolic flux in response to gene knockouts. ROOM's growth rate and flux predictions are compared with previously suggested algorithms, minimization of metabolic adjustment, and flux balance analysis (FBA). We find that minimization of metabolic adjustment provides accurate predictions for the initial transient growth rates observed during the early postperturbation state, whereas ROOM and FBA more successfully predict final higher steady-state growth rates. Although FBA explicitly maximizes the growth rate, ROOM does not, and only implicitly favors flux distributions having high growth rates. This indicates that, even though the cell has not evolved to cope with specific mutations, regulatory mechanisms aiming to minimize flux changes after genetic perturbations may indeed work to this effect. Further work is needed to identify metrics that characterize the complete trajectory from the initial to the final metabolic steady states after genetic perturbations. PMID- 15897463 TI - Sumoylation induced by the Arf tumor suppressor: a p53-independent function. AB - The mouse p19(Arf) protein has both p53-dependent and p53-independent tumor suppressive activities. Arf triggers sumoylation of many cellular proteins, including Mdm2 and nucleophosmin (NPM/B23), with which p19(Arf) physically interacts in vivo, and this occurs equally well in cells expressing or lacking functional p53. In an Arf-null NIH 3T3 cell derivative (MT-Arf cells) engineered to reexpress an Arf transgene driven by a zinc-inducible metallothionein promoter, sumoylation of endogenous Mdm2 and NPM proteins was initiated as p19(Arf) was induced and was observed before p53-dependent cell cycle arrest. Predominately nucleoplasmic molecules visualized by immunofluorescence with antibodies to small ubiquitin-like modifier (SUMO) 1 localized to nucleoli as p19(Arf) accumulated there. Two Arf mutants, one of which binds to Mdm2 and NPM but is excluded from nucleoli and the other of which enters nucleoli but is handicapped in binding to Mdm2 and NPM, were defective in inducing sumoylation of these two target proteins and did not localize bulk sumoylated molecules to nucleoli. The CELO adenovirus protein, Gam1, which inhibits the SUMO activating enzyme (E1) and leads to down-regulation of the SUMO conjugating enzyme (E2/Ubc9), had no overt effect on the ability of p19(Arf) to activate p53 or the p53-responsive genes encoding Mdm2 and p21(Cip1), despite the fact that Arf induced sumoylation of Mdm2 was blocked. Reduction of Ubc9 levels with short hairpin RNAs rendered similar results. We suggest that Arf's p53-independent effects on gene expression and tumor suppression might depend on Arf-induced sumoylation. PMID- 15897464 TI - Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to gefitinib. AB - Non-small cell lung cancers (NSCLCs) with activating mutations in the kinase domain of the epidermal growth factor receptor (EGFR) demonstrate dramatic, but transient, responses to the reversible tyrosine kinase inhibitors gefitinib (Iressa) and erlotinib (Tarceva). Some recurrent tumors have a common secondary mutation in the EGFR kinase domain, T790M, conferring drug resistance, but in other cases the mechanism underlying acquired resistance is unknown. In studying multiple sites of recurrent NSCLCs, we detected T790M in only a small percentage of tumor cells. To identify additional mechanisms of acquired resistance to gefitinib, we used NSCLC cells harboring an activating EGFR mutation to generate multiple resistant clones in vitro. These drug-resistant cells demonstrate continued dependence on EGFR and ERBB2 signaling for their viability and have not acquired secondary EGFR mutations. However, they display increased internalization of ligand-activated EGFR, consistent with altered receptor trafficking. Although gefitinib-resistant clones are cross-resistant to related anilinoquinazolines, they demonstrate sensitivity to a class of irreversible inhibitors of EGFR. These inhibitors also show effective inhibition of signaling by T790M-mutant EGFR and killing of NSCLC cells with the T790M mutation. Both mechanisms of gefitinib resistance are therefore circumvented by irreversible tyrosine kinase inhibitors. Our findings suggest that one of these, HKI-272, may prove highly effective in the treatment of EGFR-mutant NSCLCs, including tumors that have become resistant to gefitinib or erlotinib. PMID- 15897465 TI - A direct intracranial record of emotions evoked by subliminal words. AB - A classical but still open issue in cognitive psychology concerns the depth of subliminal processing. Can the meaning of undetected words be accessed in the absence of consciousness? Subliminal priming experiments in normal subjects have revealed only small effects whose interpretation remains controversial. Here, we provide a direct demonstration of semantic access for unseen masked words. In three epileptic patients with intracranial electrodes, we recorded brain potentials from the amygdala, a neural structure that responds to fearful or threatening stimuli presented in various modalities, including written words. We show that the subliminal presentation of emotional words modulates the activity of the amygdala at a long latency (>800 ms). Our result indicates that subliminal words can trigger long-lasting cerebral processes, including semantic access to emotional valence. PMID- 15897466 TI - Inhibition of protein kinase CK2 prevents the progression of glomerulonephritis. AB - Glomerulonephritis (GN) is a progressive inflammation that may be caused by a variety of underlying disorders. It is the primary cause of chronic renal failure and end-stage renal disease, which require dialysis and transplantation worldwide. Immunosuppressive therapy has been used to treat GN clinically, but this treatment has had insufficient therapeutic effects. Here, we show that protein kinase CK2 is a key molecule in the progression of GN. cDNA microarray analysis identified CK2alpha, the catalytic subunit of CK2, as a GN-related, differentially expressed gene. Overexpression of CK2alpha was noted in the proliferative glomerular lesions in rat GN models and in renal biopsy specimens from lupus nephritis or IgA nephropathy patients. Administration of either antisense oligodeoxynucleotide against CK2alpha or low molecular weight CK2 specific inhibitors effectively prevented the progression of renal pathology in the rat GN models. The resolution of GN by CK2 inhibition may result from its suppression of extracellular signal-regulated kinase-mediated cell proliferation, and its suppression of inflammatory and fibrotic processes that are enhanced in GN. Our results show that CK2 plays a critical role in the progression of immunogenic renal injury, and therefore, CK2 is a potential target for GN therapy. PMID- 15897467 TI - Human coronavirus NL63 employs the severe acute respiratory syndrome coronavirus receptor for cellular entry. AB - Coronavirus (CoV) infection of humans is usually not associated with severe disease. However, discovery of the severe acute respiratory syndrome (SARS) CoV revealed that highly pathogenic human CoVs (HCoVs) can evolve. The identification and characterization of new HCoVs is, therefore, an important task. Recently, a HCoV termed NL63 was discovered in patients with respiratory tract illness. Here, cell tropism and receptor usage of HCoV-NL63 were analyzed. The NL63 spike (S) protein mediated infection of different target cells compared with the closely related 229E-S protein but facilitated entry into cells known to be permissive to SARS-CoV-S-driven infection. An analysis of receptor engagement revealed that NL63-S binds angiotensin-converting enzyme (ACE) 2, the receptor for SARS-CoV, and HCoV-NL63 uses ACE2 as a receptor for infection of target cells. Potent neutralizing activity directed against NL63- but not 229E-S protein was detected in virtually all sera from patients 8 years of age or older, suggesting that HCoV NL63 infection of humans is common and usually acquired during childhood. Here, we show that SARS-CoV shares its receptor ACE2 with HCoV-NL63. Because the two viruses differ dramatically in their ability to induce disease, analysis of HCoV NL63 might unravel pathogenicity factors in SARS-CoV. The frequent HCoV-NL63 infection of humans suggests that highly pathogenic variants have ample opportunity to evolve, underlining the need for vaccines against HCoVs. PMID- 15897468 TI - Dimeric SecA is essential for protein translocation. AB - SecA facilitates bacterial protein translocation by its association with presecretory or membrane proteins and the SecYEG translocon channel. Once assembled, SecA ATPase undergoes cycles of membrane insertion and retraction at SecYEG that drive protein translocation in a stepwise fashion. SecA exists in equilibrium between a monomer and dimer, and association with its translocation ligands shifts this equilibrium dramatically. Here, we examined the proposal that protein translocation can occur by means of a SecA monomer. We produced a mutant SecA protein lacking residues 2-11, which was found to exist mostly as a monomer, and it was unable to complement a conditional-lethal secA mutant, was inactive for in vitro protein translocation, and was poorly active for translocation ATPase activity. Furthermore, we developed a technique termed membrane trapping, where wild-type SecA subunits became trapped within the membrane by overproduction of membrane-stuck mutant SecA proteins, and, in one case, a membrane-associated SecA heterodimer was demonstrated. Finally, we examined both endogenous and reconstituted membrane-bound SecA and found a significant level of SecA dimer in both cases, as assessed by chemical crosslinking. Collectively, our results strongly suggest that membrane-bound SecA dimer is critical for the protein translocation cycle, although these results cannot exclude participation of SecA monomer at some stage in the translocation process. Our findings have important implications regarding SecA motor function and translocon assembly and activation. PMID- 15897469 TI - Stable X chromosome inactivation involves the PRC1 Polycomb complex and requires histone MACROH2A1 and the CULLIN3/SPOP ubiquitin E3 ligase. AB - X inactivation involves the stable silencing of one of the two X chromosomes in XX female mammals. Initiation of this process occurs during early development and involves Xist (X-inactive-specific transcript) RNA coating and the recruitment of Polycomb repressive complex (PRC) 2 and PRC1 proteins. This recruitment results in an inactive state that is initially labile but is further locked in by epigenetic marks such as DNA methylation, histone hypoacetylation, and MACROH2A deposition. Here, we report that the E3 ubiquitin ligase consisting of SPOP and CULLIN3 is able to ubiquitinate the Polycomb group protein BMI1 and the variant histone MACROH2A. We find that in addition to MACROH2A, PRC1 is recruited to the inactivated X chromosome in somatic cells in a highly dynamic, cell cycle regulated manner. Importantly, RNAi-mediated knock-down of CULLIN3 or SPOP results in loss of MACROH2A1 from the inactivated X chromosome (Xi), leading to reactivation of the Xi in the presence of inhibitors of DNA methylation and histone deacetylation. Likewise, Xi reactivation is also seen on MacroH2A1 RNAi under these conditions. Hence, we propose that the PRC1 complex is involved in the maintenance of X chromosome inactivation in somatic cells. We further demonstrate that MACROH2A1 deposition is regulated by the CULLIN3/SPOP ligase complex and is actively involved in stable X inactivation, likely through the formation of an additional layer of epigenetic silencing. PMID- 15897470 TI - A network analysis of committees in the U.S. House of Representatives. AB - Network theory provides a powerful tool for the representation and analysis of complex systems of interacting agents. Here, we investigate the U.S. House of Representatives network of committees and subcommittees, with committees connected according to "interlocks," or common membership. Analysis of this network reveals clearly the strong links between different committees, as well as the intrinsic hierarchical structure within the House as a whole. We show that network theory, combined with the analysis of roll-call votes using singular value decomposition, successfully uncovers political and organizational correlations between committees in the House without the need to incorporate other political information. PMID- 15897471 TI - Psychosocial and behavioral aspects of bariatric surgery. AB - Bariatric surgery has become an increasingly popular treatment option for individuals with extreme obesity (defined as a BMI > or = 40 kg/m2) or those with less severe obesity accompanied by significant comorbidities. Sustained postoperative weight loss and improvements in obesity-related health problems make bariatric surgery the most effective treatment for this population. Nevertheless, most experts agree that psychosocial and behavioral factors contribute to successful postoperative outcomes. This paper reviews the literature on the preoperative psychosocial status, eating behaviors, and quality of life of patients who seek bariatric surgery. In addition, the paper examines studies that investigated changes in these factors postoperatively. The review concludes with an agenda for future research in this area. PMID- 15897472 TI - Evaluation of reference genes for studies of gene expression in human adipose tissue. AB - OBJECTIVE: The aim of this study was to evaluate reference genes for expression studies of human adipose tissue. RESEARCH METHODS AND PROCEDURES: Using 52 human adipose tissue expression profiles (HU95), 10 putative reference genes with the lowest variation in expression levels were selected for further studies. Expression stability of these 10 novel and 5 previously established reference genes was evaluated by real-time reverse transcriptase-polymerase chain reaction analysis. For this purpose, 44 adipose tissue biopsies from 27 subjects were chosen to include a wide range of parameters such as sex, age, BMI, depot origin, biopsy procedure, and effects of nutrition. RESULTS: LRP10 was identified as the gene with the least variation in expression levels. The frequently used reference genes RPLP0, 18S rRNA, PPIA, ACTB, and GAPD were ranked as 4, 6, 7, 8, and 10, respectively. DISCUSSION: Our results suggest that LRP10 is a better choice as reference for expression studies of human adipose tissue compared with the most frequently used reference genes. PMID- 15897473 TI - Neuropeptides, including neuropeptide Y and melanocortins, mediate lipolysis in murine adipocytes. AB - OBJECTIVE: To determine whether key appetite-regulating neuropeptides such as melanin-concentrating hormone (MCH), neuropeptide Y (NPY), and alpha-melanocyte stimulating hormone (alpha-MSH), which are known to mediate energy balance through centrally mediated pathways, also have direct acute effects on the lipolytic activity of murine adipocytes. RESEARCH METHODS AND PROCEDURES: Fully differentiated 3T3-L1 adipocytes serum starved overnight in Dulbecco's modified Eagle medium containing 2% bovine serum albumin or freshly isolated mouse adipocytes were incubated for up to 2 hours in the absence and presence of 100 nM each of NPY, MCH, alpha-MSH, the melanocortin receptor agonist MTII, or isoproterenol as a control. Free fatty acids secreted into the incubation medium were measured using a commercially available nonesterified fatty acid C test kit. RESULTS: Treatment of 3T3-L1 cells with 100 nM NPY decreased basal free fatty acid secretion (basal, 0.006 +/- 0.001 vs. NPY, 0.001 +/- 0.0003 nM at 90 minutes; p < 0.05), whereas both alpha-MSH and MTII stimulated up to a 7-fold increase in free fatty acid release (MTII, 0.238 +/- 0.004 vs. basal, 0.024 +/- 0.002 nM at 2 hours; p < 0.05; and alpha-MSH, 0.22 +/- 0.005 vs. basal, 0.04 +/- 0.003 nM at 2 hours; p < 0.05). Treatment with 100 nM MCH had no effect on basal free fatty acid release or on alpha-MSH-induced lipolysis during concurrent treatment. Conversely, concurrent treatment with 100 nM NPY dramatically inhibited (by approximately 90%) alpha-MSH-induced lipolysis. Similar treatment of freshly isolated mouse adipocytes showed virtually identical results. DISCUSSION: In addition to their centrally mediated actions, appetite-regulating neuropeptides modulate adipose tissue mass through direct peripheral effects. Systemic administration of pharmacological agents altering the effects of these neuropeptides may form the basis of future obesity therapies. Thus, some of these agents will likely have direct effects on adipocytes that may serve to alter their therapeutic effectiveness. PMID- 15897474 TI - Regulation of adiponectin expression in human adipocytes: effects of adiposity, glucocorticoids, and tumor necrosis factor alpha. AB - OBJECTIVE: Adiponectin mRNA expression in isolated subcutaneous and omental adipocytes was examined across a wide range of adiposity to determine whether adiponectin synthesis is impaired in these adipose tissue depots in obese humans. Tumor necrosis factor (TNF)alpha and dexamethasone were tested for inhibitory effects on adiponectin release from human adipocytes in vitro. RESEARCH METHODS AND PROCEDURES: Adipocytes were isolated by collagenase digestion of abdominal adipose tissue obtained from subjects undergoing surgical procedures or outpatient needle biopsy. Adiponectin and leptin mRNA were quantitated by real time reverse transcriptase-polymerase chain reaction. Adiponectin and leptin secretion from isolated adipocytes treated with dexamethasone or TNFalpha were determined by radioimmunoassay. RESULTS: There was a significant negative correlation between adiponectin gene expression and BMI in subcutaneous adipocytes from 32 women (r = 0.420; p = 0.02). Adiponectin mRNA was also significantly correlated with serum adiponectin (r = 0.44; p = 0.03; n = 25). There was no correlation between adiponectin mRNA expression and BMI in omental adipocytes from 29 women. Leptin mRNA was significantly and positively correlated (r = 0.484; p = 0.01) with BMI in the same omental adipocyte mRNA preparations. In subcutaneous adipocytes from lean subjects, TNFalpha inhibited adiponectin release by 7.4 +/- 1.2% (n = 9, p < 0.05) but had no effect on adiponectin release from subcutaneous or omental adipocytes from obese subjects. Dexamethasone significantly inhibited adiponectin release with 24 hours of treatment. DISCUSSION: The data suggest that reduced adiponectin synthesis in subcutaneous adipocytes contributes to lower serum adiponectin levels in obesity and that glucocorticoids regulate adiponectin gene expression in human adipocytes. TNFalpha does not seem to directly inhibit adiponectin synthesis in human adipocytes. PMID- 15897475 TI - 1,25-dihydroxyvitamin D3 modulation of adipocyte glucocorticoid function. AB - OBJECTIVE: 1,25-Dihydroxyvitamin D3 dose dependently increases intracellular calcium in human adipocytes. We have demonstrated that suppression of circulating 1,25-dihydroxyvitamin D3 levels by increasing dietary calcium reduces adipocyte intracellular calcium and reduces adiposity in both humans and rodents, with preferential loss of trunk fat. Autocrine production of cortisol by adipocytes of mice overexpressing 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD 1) in adipose tissue increases visceral adiposity, whereas knockout of 11beta-HSD 1 appears to attenuate truncal obesity. Accordingly, our objective was to investigate the role of 1,25-dihydroxyvitamin D3 in the modulation of adipocyte glucocorticoid metabolism. RESEARCH METHODS AND PROCEDURES: We examined the effect of 1,25-dihydroxyvitamin D3 or angiotensin II on cortisol production and expression using real-time reverse transcriptase-polymerase chain reaction of 11beta-HSD 1, angiotensin II receptor type 1 (AT1), and AT2 receptor in human adipocytes. RESULTS: Adipocytes produced negligible cortisol in the absence of substrate (cortisone). In the presence of cortisone (1 to 10 nM), there was significant cortisol production, which was dose dependently augmented (2- to 6 fold, p < 0.001) by 1,25-dihydroxyvitamin D3 (0.1 to 10 nM). 1,25 Dihydroxyvitamin D3 dose dependently increased 11beta-HSD 1 expression up to 2 fold (p < 0.01) in both the presence and absence of cortisone. In contrast, 1,25 dihydroxyvitamin D3 dose dependently decreased adipocyte AT1 expression (by 30% to 50%, p < 0.001) in both the presence and absence of cortisone, suggesting compensatory down-regulation of AT(1). DISCUSSION: We conclude that 1,25 dihydroxyvitamin D3 directly regulates adipocyte 11beta-HSD 1 expression and, consequently, local cortisol levels and that this may contribute to the preferential loss of visceral adiposity by high-calcium diets. PMID- 15897476 TI - Down-regulation of pyruvate dehydrogenase phosphatase in obese subjects is a defect that signals insulin resistance. AB - OBJECTIVE: The objective of this study was to determine whether down-regulation of pyruvate dehydrogenase phosphatase (PDP) is responsible for poorly active pyruvate dehydrogenase (PDH) in circulating lymphocytes (CLs) of obese subjects (ObS), and if so, whether it improves when their plasma insulin rises. RESEARCH METHODS AND PROCEDURES: PDH activity was compared in lysed CLs of 10 euglycemic ObS and 10 sex- and age-matched controls before and during plasma insulin enhancement in an oral glucose tolerance test. It was evaluated without (PDHa) or with Mg/Ca or Mg at various concentrations to assess PDP1 or PDP2 activities or with Mg/Ca and exogenous PDP to determine total PDH activity (PDHt), which is an indirect measure of the amount of PDH. The insulin sensitivity index was calculated, and PDP1 and PDP2 mRNA was sought in the CLs. RESULTS: At T0 in ObS, PDHt was normal, whereas PDHa and PDP1 activity was below normal at all Mg/Ca concentrations. PDP2 activity was undetectable in both groups. PDP1 and PDP2 mRNA was identified, and insulin sensitivity index and PDHa were directly correlated. During the oral glucose tolerance test, plasma insulin rose considerably more in ObS than in controls; PDHa and PDP1 activity also increased but remained significantly below normal, and PDHt was unvaried in both groups. DISCUSSION: PDP1 is down-regulated in CLs of ObS because it is poorly sensitive to Mg/Ca; this defect is attenuated when plasma insulin is greatly enhanced. PMID- 15897477 TI - Prenatal exposure to nicotine causes postnatal obesity and altered perivascular adipose tissue function. AB - OBJECTIVE: Recent epidemiological studies have shown that there is an increased risk of obesity and hypertension in children born to women who smoked during pregnancy. The aim of this study was to examine the effect of fetal and neonatal exposure to nicotine, the major addictive component of cigarette smoke, on postnatal adiposity and blood vessel function. RESEARCH METHODS AND PROCEDURES: Female Wistar rats were given nicotine or saline (vehicle) during pregnancy and lactation. Postnatal growth was determined in the male offspring from weaning until 26 weeks of age. At 26 weeks of age, fat pad weight and the function of the perivascular adipose tissue (PVAT) in the thoracic aorta and mesenteric arteries were examined. RESULTS: Exposure to nicotine resulted in increased postnatal body weight and fat pad weight and an increased amount of PVAT in the offspring. Contraction of the aorta induced by phenylephrine was significantly attenuated in the presence of PVAT, whereas this effect was not observed in the aortic rings from the offspring of nicotine-exposed dams. Phenylephrine-induced contraction without PVAT was not different between saline- and nicotine-exposed rats. Transfer of solution incubated with PVAT-intact aorta to PVAT-free aorta induced a marked relaxation response in the rats from saline-exposed dams, but this relaxation response was significantly impaired in the rats from nicotine-exposed dams. DISCUSSION: Our results showed that prenatal nicotine exposure increased adiposity and caused an alteration in the modulatory function of PVAT on vascular relaxation response, thus providing insight into the mechanisms underlying the increased prevalence of obesity and hypertension in children exposed to cigarette smoke in utero. PMID- 15897478 TI - Intentional weight loss reduces mortality rate in a rodent model of dietary obesity. AB - OBJECTIVE: We used a rodent model of dietary obesity to evaluate effects of caloric restriction-induced weight loss on mortality rate. Research Measures and Procedures: In a randomized parallel-groups design, 312 outbred Sprague-Dawley rats (one-half males) were assigned at age 10 weeks to one of three diets: low fat (LF; 18.7% calories as fat) with caloric intake adjusted to maintain body weight 10% below that for ad libitum (AL)-fed rat food, high fat (HF; 45% calories as fat) fed at the same level, or HF fed AL. At age 46 weeks, the lightest one-third of the AL group was discarded to ensure a more obese group; the remaining animals were randomly assigned to one of three diets: HF-AL, HF with energy restricted to produce body weights of animals restricted on the HF diet throughout life, or LF with energy restricted to produce the body weights of animals restricted on the LF diet throughout life. Life span, body weight, and leptin levels were measured. RESULTS: Animals restricted throughout life lived the longest (p < 0.001). Life span was not different among animals that had been obese and then lost weight and animals that had been nonobese throughout life (p = 0.18). Animals that were obese and lost weight lived substantially longer than animals that remained obese throughout life (p = 0.002). Diet composition had no effect on life span (p = 0.52). DISCUSSION: Weight loss after the onset of obesity during adulthood leads to a substantial increase in longevity in rats. PMID- 15897479 TI - Insulin sensitivity determines the effectiveness of dietary macronutrient composition on weight loss in obese women. AB - OBJECTIVE: To determine whether macronutrient composition of a hypocaloric diet can enhance its effectiveness and whether insulin sensitivity (Si) affects the response to hypocaloric diets. RESEARCH METHODS AND PROCEDURES: Obese nondiabetic insulin-sensitive (fasting insulin < 10 microU/mL; n = 12) and obese nondiabetic insulin-resistant (fasting insulin > 15 microU/mL; n = 9) women (23 to 53 years old) were randomized to either a high carbohydrate (CHO) (HC)/low fat (LF) (60% CHO, 20% fat) or low CHO (LC)/high fat (HF) (40% CHO, 40% fat) hypocaloric diet. Primary outcome measures after a 16-week dietary intervention were: changes in body weight (BW), Si, resting metabolic rate, and fasting lipids. RESULTS: Insulin-sensitive women on the HC/LF diet lost 13.5 +/- 1.2% (p < 0.001) of their initial BW, whereas those on the LC/HF diet lost 6.8 +/- 1.2% (p < 0.001; p < 0.002 between the groups). In contrast, among the insulin-resistant women, those on the LC/HF diet lost 13.4 +/- 1.3% (p < 0.001) of their initial BW as compared with 8.5 +/- 1.4% (p < 0.001) lost by those on the HC/LF diet (p < 0.04 between two groups). These differences could not be explained by changes in resting metabolic rate, activity, or intake. Overall, changes in Si were associated with the degree of weight loss (r = -0.57, p < 0.05). DISCUSSION: The state of Si determines the effectiveness of macronutrient composition of hypocaloric diets in obese women. For maximal benefit, the macronutrient composition of a hypocaloric diet may need to be adjusted to correspond to the state of Si. PMID- 15897480 TI - Decreased glucagon-like peptide 1 release after weight loss in overweight/obese subjects. AB - OBJECTIVE: Postprandial glucagon-like peptide 1 (GLP-1) release seems to be attenuated in obese subjects. Results on whether weight loss improves GLP-1 release are contradictory. The aim of this study was to further investigate the effect of weight loss on basal and postprandial GLP-1 release in overweight/obese subjects. RESEARCH METHODS AND PROCEDURES: Thirty-two overweight/obese subjects participated in a repeated measurement design before (BMI, 30.3 +/- 2.8 kg/m2; waist circumference, 92.6 +/- 7.8 cm; hip circumference, 111.1 +/- 7.4 cm) and after a weight loss period of 6 weeks (BMI, 28.2 +/- 2.7 kg/m2; waist circumference, 85.5 +/- 8.5 cm; hip circumference, 102.1 +/- 9.2 cm). During weight loss, subjects received a very-low-calorie diet (Optifast) to replace three meals per day. Subjects came to the laboratory fasted, and after a baseline blood sample, received a standard breakfast (1.9 MJ). Postprandially, blood samples were taken every one-half hour relative to intake for 120 minutes to determine GLP-1, insulin, glucose, and free fatty acids from plasma. Appetite ratings were obtained with visual analog scales. RESULTS: After weight loss, postprandial GLP-1 concentrations at 30 and 60 minutes were significantly lower than before weight loss (p < 0.05). Glucose concentrations were also lower, and free fatty acids were higher compared with before weight loss. Ratings of satiety were increased, and hunger scores were decreased after weight loss (p < 0.05). DISCUSSION: In overweight/obese subjects, GLP-1 concentrations after weight loss were decreased compared with before weight loss, and nutrient-related stimulation was abolished. This might be a response to a proceeding negative energy balance. Satiety and GLP-1 seem to be unrelated in the long term. PMID- 15897481 TI - Dysregulation of the autonomic nervous system can be a link between visceral adiposity and insulin resistance. AB - OBJECTIVE: To evaluate the interplay among abdominal adipose tissue distribution, the cortisol axis, the autonomic nervous system, and insulin resistance. RESEARCH METHODS AND PROCEDURES: Two age-, sex-, and BMI-matched groups were studied. Fifteen subjects were first-degree relatives of patients with type 2 diabetes (R), and 15 had no family history of diabetes (controls, C). A hyperinsulinemic euglycemic clamp, cortisol measurements, and analysis of heart rate variability (HRV) were performed. Computed tomography was performed in a subgroup (n = 9 + 9) to determine abdominal adipose tissue distribution. RESULTS: R tended to be less insulin-sensitive than C (M value 9.2 +/- 1.0 vs 10.3 +/- 0.7 mg/kg per minute, not significant). Stimulation with tetracosactin or corticotropin releasing hormone yielded lower peak serum cortisol levels in R (p = 0.03 and p = 0.06, respectively). The amount of visceral abdominal fat (VAT) tended to be greater in R. In all subjects, VAT was negatively correlated to insulin sensitivity (r = 0.93, p < 0.001). There was a positive association between VAT and resting heart rate (r = 0.70, p = 0.003) and sympathetic/parasympathetic ratio in HRV assessment after tilt (r = 0.53, p = 0.03). Subcutaneous abdominal tissue was not associated with insulin sensitivity or any of the hormonal or HRV assessments. DISCUSSION: Subjects genetically predisposed for type 2 diabetes had a tendency toward a larger amount of VAT and to lower insulin sensitivity compared with control subjects. The amount of visceral fat was strongly associated with insulin resistance and signs of a high ratio of sympathetic vs. parasympathetic reactivity. A large amount of visceral fat may act in concert with sympathetic/parasympathetic imbalance to promote the development of insulin resistance, and this may be partly independent of genetic background. PMID- 15897482 TI - Prediction of percentage body fat in rural thai population using simple anthropometric measurements. AB - OBJECTIVE: To develop and validate sex-specific equations for predicting percentage body fat (%BF) in rural Thai population, based on BMI and anthropometric measurements. RESEARCH METHODS AND PROCEDURES: %BF (DXA; GE Lunar Corp., Madison, WI) was measured in 181 men and 255 women who were healthy and between 20 and 84 years old. Anthropometric measures such as weight (kilograms), height (centimeters), BMI (kilograms per meter squared), waist circumference (centimeters), hip circumference (centimeters), thickness at triceps skinfold (millimeters), biceps skinfold (millimeters), subscapular skinfold (millimeters), and suprailiac skinfold (millimeters) were also measured. The sample was randomly divided into a development group (98 men and 125 women) and a validation group (83 men and 130 women). Regression equations of %BF derived from the development group were then evaluated for accuracy in the validation group. RESULTS: The equation for estimating %BF in men was: %BF(men) = 0.42 x subscapular skinfold + 0.62 x BMI - 0.28 x biceps skinfold + 0.17 x waist circumference - 18.47, and in women: %BF(women) = 0.42 x hip circumference + 0.17 x suprailiac skinfold + 0.46 x BMI - 23.75. The coefficient of determination (R2) for both equations was 0.68. Without anthropometric variables, the predictive equation using BMI, age, and sex was: %BF = 1.65 x BMI + 0.06 x age - 15.3 x sex - 10.67 (where sex = 1 for men and sex = 0 for women), with R2 = 0.83. When these equations were applied to the validation sample, the difference between measured and predicted %BF ranged between +/-9%, and the positive predictive values were above 0.9. DISCUSSION: These results suggest that simple, noninvasive, and inexpensive anthropometric variables may provide an accurate estimate of %BF and could potentially aid the diagnosis of obesity in rural Thais. PMID- 15897483 TI - Ghrelin does not influence gastric emptying in obese subjects. AB - OBJECTIVE: To evaluate the relationship between fasting plasma concentrations of ghrelin and gastric emptying in obese individuals compared with lean subjects. RESEARCH METHODS AND PROCEDURES: We included 20 obese patients (9 men and 11 women, BMI > 30 kg/m2) and 16 nonobese control subjects (7 men and 9 women, BMI < or = 25 kg/m2). Gastric emptying of solids (egg sandwich labeled with radionuclide) was measured at 120 minutes with (99m)Tc-single photon emission computed tomography imaging. Ghrelin and leptin were analyzed by radioimmunoassay and ELISA methods, respectively. RESULTS: The gastric half-emptying time was similar in obese men and women (67.8 +/- 14.79 vs. 66.6 +/- 13.56 minutes) but significantly shorter (p < 0.001) than in the control population (men: 88.09 +/- 11.72 minutes; women: 97.25 +/- 10.31 minutes). Ghrelin levels were significantly lower in obese subjects (131.37 +/- 47.67 vs. 306.3 +/- 45.52 pg/mL; p < 0.0001 in men and 162.13 +/- 32.95 vs. 272.8 +/- 47.77 pg/mL; p < 0.0001 in women). A negative correlation between gastric emptying and fasting ghrelin levels was observed only in lean subjects (y = -0.2391x + 157.9; R2 = 0.95). Also, in the lean group, ghrelin was the only significant independent determinant of gastric emptying, explaining 98% of the variance (adjusted R2) in a multiple regression analysis. DISCUSSION: This report shows that, in humans, gastric emptying is faster in obese subjects than in lean controls and that, whereas ghrelin is the best determinant of gastric kinetics in healthy controls, this action is lost in obesity. PMID- 15897484 TI - Intentional weight loss in young adults: sex-specific genetic and environmental effects. AB - OBJECTIVE: To explore eating styles associated with intentional weight loss (IWL) and to determine whether the genetic liability in IWL is entirely shared with genetic liability affecting BMI. RESEARCH METHODS AND PROCEDURES: As part of a longitudinal assessment of various health-related behaviors in a large population based sample of twins, eating styles, BMI, and the number of times the study participants had intentionally lost > or = 5 kg were assessed by questionnaire from 4667 male and female twins (22 to 27 years of age). Associations of eating styles and IWL were explored using polytomous logistic regression models adjusted for BMI. Sex-specific bivariate structural equation modeling was used to explore genetic and environmental correlations of BMI and IWL. RESULTS: Individuals who had engaged in IWL exhibited markedly more restricting, overeating, and alternating restricting/overeating than those in the no-IWL group. Snacking and eating in the evening were characteristic of women with at least two IWL attempts. Eating in response to visual and emotional cues was very pronounced in women who had engaged in IWL but much less so in men. IWL was estimated to have a heritability of 38% [95% confidence interval (CI), 19% to 55%] in men and 66% (95% CI, 55% to 75%) in women. The genetic covariance of BMI and IWL was 0.38 (95% CI, 0.28 to 0.47) for men and 0.45 (95% CI, 0.41 to 0.52) for women. DISCUSSION: Distinct sex differences exist in eating styles associated with IWL and in the heritability of IWL. Most genetic factors affecting BMI are different from those affecting IWL. PMID- 15897485 TI - Survival expectations of the obese: Is excess mortality reflected in perceptions? AB - OBJECTIVE: This study compared self-reported subjective life expectancy (i.e., probability of living to age 75) for normal-weight, overweight, and obese weight groups to examine whether individuals are internalizing information about the health risks due to excessive weight. RESEARCH METHODS AND PROCEDURES: Using data from the Health and Retirement Study, a total of 9035 individuals 51 to 61 years old were analyzed by BMI category. The primary outcome measure was individuals' reports about their own expectations of survival to age 75. Absolute and relative risks of survival were compared with published estimates of survival to age 75. RESULTS: Consistently, higher levels of BMI were associated with lower self estimated survival probabilities. Differences relative to normal weight ranged from 4.9% (p < 0.01) for male nonsmokers to 8.8% (p < 0.001) for female nonsmokers. However, these differences were substantially less than those obtained from published survival curve estimates, suggesting that obese individuals tended to underestimate mortality risks. DISCUSSION: Individuals appeared to underestimate the mortality risks of excessive weight; thus, knowledge campaigns about the risks of obesity should remain a top priority. PMID- 15897486 TI - Interactive multimedia for promoting physical activity (IMPACT) in children. AB - OBJECTIVE: To develop and examine the efficacy of a computer-based interactive multimedia curriculum for promoting physical activity in fourth grade children. RESEARCH METHODS AND PROCEDURES: The participants were 209 fourth grade children (mean age of 9.5 +/- 0.4 years) from four schools. Two schools received an 8-week multimedia intervention delivered by interactive CD-ROM, supplemented by four classroom and four homework assignments. Two control schools received educational CD-ROMS not related to health outcomes. Measures conducted before and after intervention included height, weight, percentage body fat (bioimpedance analysis), physical activity (5-day accelerometry), and psychosocial aspects of physical activity by questionnaire. All outcomes were examined using general linear models. RESULTS: There was a significant treatment effect for obesity reduction in girls but not in boys. There were no significant treatment effects on total physical activity by accelerometry (total counts per minute), but there was an overall treatment effect on reducing percent of time in moderate-intensity activity (16.5% to 15% of the time) and significant sex-by- treatment interactions for light-intensity activities (reduction in boys from 78% to 75% of the time and an increase in girls from 78% to 81% of the time). There were marginal/significant treatment effects for improvements in behavioral outcomes, including self-efficacy (p = 0.06), social norms (p = 0.07), and outcome expectancies (p = 0.049). DISCUSSION: The interactive multimedia curriculum favored an improvement in obesity indices in girls and was associated with subtle changes in physical activity in girls and general improvement in psychosocial outcomes related to physical activity. PMID- 15897487 TI - Obesity-related comorbidities in obese African Americans in an outpatient weight loss program. AB - OBJECTIVE: To identify, among obese African-American enrollees in an outpatient weight loss program, differences between those with and without obesity-related comorbidities (ORCMs). RESEARCH METHODS AND PROCEDURES: Data were from 237 obese African Americans (BMI, 30 to 50 kg/m2; 90% women) who enrolled in a 10-week lifestyle weight loss program. Analyses compared subgroups defined by ORCM status (from medical history) on baseline characteristics, program attendance, and postprogram weight change. RESULTS: Most participants (76%) had one or more ORCMs. Those with versus without ORCMs, respectively, were older (mean age, 45.6 vs. 37.1 years; p < 0.001), were less educated (59.2% vs. 76.6% with > 12 years; p = 0.031), were more likely to perceive a physical limitation affecting activity (22.2% vs. 1.8%; p < 0.001), and had higher waist circumference (mean, 113.7 vs. 106.9 cm; p < 0.001) but not BMI (38.3 vs. 37.0 kg/m2; p = 0.095). Logistic regression analyses confirmed the independence of these associations. Having ORCMs was not associated with class attendance or return for data collection after the 10-week program. Postprogram weight change (n = 134) was unrelated to ORCMs, but better weight loss was seen among those without perceived physical limitations (1.9 vs. 0.4 kg in those without versus with limitations; p = 0.069). CONCLUSION: Data from this clinical sample of obese African Americans suggest that waist circumference is relevant to ORCM status at BMI levels up to 50 kg/m2. Clear indications for tailoring of treatment based on ORCM status were not identified, although the possible influence of ORCM-related activity limitations warrants further study. PMID- 15897488 TI - Altered luteinizing hormone and testosterone secretion in middle-aged obese men with obstructive sleep apnea. AB - OBJECTIVE: To elucidate the causes for the decline in testosterone levels observed in men with obstructive sleep apnea (OSA). RESEARCH METHODS AND PROCEDURES: We determined serum luteinizing hormone (LH) and testosterone levels every 20 minutes between 7 pm and 7 am with simultaneous sleep recordings in five obese middle-aged men with OSA, in five age- and BMI-matched controls, and in six lean young healthy men. RESULTS: The mean and area under the curve (AUC) values of LH and testosterone were significantly lower in men with OSA compared with controls. Young controls had significantly more testosterone pulses of shorter interpulse duration than OSA subjects and middle-aged controls. After adjusting for age and BMI, the three groups differed in mean and AUC values of LH and testosterone. Analysis of covariance, using BMI as a covariate, revealed a statistically significant group effect on mean and AUC testosterone values (p = 0.03; p < 0.003, respectively). Eliminating young controls, there was a significant positive correlation between the amount of LH and testosterone secreted at night. After partialling out age alone and BMI alone, the mean LH and mean testosterone were still positively correlated. DISCUSSION: Thus, OSA is associated with decreased pituitary-gonadal function. The decline in testosterone concentrations is due to obesity and advanced age and to a lesser degree to sleep fragmentation and hypoxia. PMID- 15897489 TI - Overweight and obesity: knowledge, attitudes, and practices of general practitioners in france. AB - OBJECTIVE: To describe the current knowledge, attitudes, and practices of French general practitioners (GPs) in the field of adult overweight and obesity management. RESEARCH METHODS AND PROCEDURES: A cross-sectional telephone survey interviewed a sample of 600 GPs, representative of the private GPs in southeastern France. A four-part questionnaire assessed personal and professional characteristics, attitudes and opinions about overweight and obesity, relevant knowledge and training, and practices (diagnostic methods, clinical assessments, weight loss objectives, types of counseling). RESULTS: Most GPs knew that weight problems are health-threatening, and 79% agreed that managing these problems is part of their role. Nevertheless, 58% did not feel they perform this role effectively, and one-third did not find it professionally gratifying. Approximately 30% had negative attitudes toward overweight and obese patients; 57% were pessimistic about patients' ability to lose weight; 64% often set weight loss objectives more demanding than guidelines call for; and neither food diaries nor nutritional education were used systematically. GPs' feelings of effectiveness and attitudes toward obese patients were associated with some professional (training) and personal (BMI, personal diet experience) characteristics. DISCUSSION: GPs' feelings of ineffectiveness may stem from an underlying conflict between practitioners' and patients' representations of weight problems and the relationship problems this causes. Inadequate practices and health care system organization may also play a role. PMID- 15897490 TI - Beyond the holy grail of motor symptoms: deep brain stimulation for Parkinson's disease. PMID- 15897491 TI - Advanced online publication. PMID- 15897492 TI - Stroke services, stroke networks: is there an ideal model? PMID- 15897493 TI - Long lasting antalgic effects of daily repetitive transcranial magnetic stimulation in neuropathic pain. PMID- 15897494 TI - Challenges for patient organisations: focus on Europe. PMID- 15897495 TI - Driver landmark and traffic sign identification in early Alzheimer's disease. AB - OBJECTIVE: To assess visual search and recognition of roadside targets and safety errors during a landmark and traffic sign identification task in drivers with Alzheimer's disease. METHODS: 33 drivers with probable Alzheimer's disease of mild severity and 137 neurologically normal older adults underwent a battery of visual and cognitive tests and were asked to report detection of specific landmarks and traffic signs along a segment of an experimental drive. RESULTS: The drivers with mild Alzheimer's disease identified significantly fewer landmarks and traffic signs and made more at-fault safety errors during the task than control subjects. Roadside target identification performance and safety errors were predicted by scores on standardised tests of visual and cognitive function. CONCLUSIONS: Drivers with Alzheimer's disease are impaired in a task of visual search and recognition of roadside targets; the demands of these targets on visual perception, attention, executive functions, and memory probably increase the cognitive load, worsening driving safety. PMID- 15897496 TI - Functional improvement after subthalamic stimulation in Parkinson's disease: a non-equivalent controlled study with 12-24 month follow up. AB - OBJECTIVE: This study aimed to assess the effectiveness of chronic bilateral STN S in improving the functional status of PD patients compared with patients treated with drugs alone. METHODS: Controlled study of disability index changes over 12 and 24 month chronic STN stimulation. Of 39 patients with advanced PD meeting CAPSIT criteria for STN-S, 23 underwent surgery; 16 patients decided against surgery and continued on drug schedule adjustments. Functional status was measured using the Activities of Daily Living section of the Unified Parkinson's Disease Rating Scale (UPDRS-ADL), Brown's Disability Scale, and Functional Independence Measure. UPDRS motor score and subscores for selected items, levodopa equivalent daily dose, and Beck Depression Inventory scores were also monitored. RESULTS: T12 follow up data were available for all 39 patients and T24 data for 13 STN-S and 8 control subjects. Compared with controls, STN-S patients experienced significant or highly significant improvements in all independence measures at both 12 and 24 months (time x treatment effect T12: F = 19.5, p = 0.00008; T24: F = 6.2, p = 0.005). Forward stepwise regression for independent predictors of the yearly rate of UPDRS-ADL score modification in the entire sample showed that treatment was the only factor significantly associated with functional status change (beta coefficient -0.54, t value -2.5, p = 0.02), whereas other variables-UPDRS motor score, BDI, and age at disease onset and enrolment-were not in the equation. CONCLUSION: STN-S is an effective therapeutic option in advanced PD. It induced a consistent improvement of functional abilities over two years to an extent that was not achieved with drug therapy alone. PMID- 15897497 TI - Does bilateral stimulation of the subthalamic nucleus aggravate apathy in Parkinson's disease? AB - OBJECTIVE: High frequency stimulation of the subthalamic nucleus (STN) dramatically decreases motor disability in patients with Parkinson"s disease (PD), but has been reported to aggravate apathy. The aim of this study was to analyse the effect of STN stimulation on motivation and reward sensitivity in a consecutive series of PD patients. METHODS: Apathy and reward sensitivity (Apathy Scale, Stimulus-Reward Learning, Reversal, Extinction, and Gambling tasks) were assessed in 18 PD patients treated by bilateral STN stimulation ("on" and "off" conditions) compared with 23 matched patients undergoing long term treatment with levodopa ("on" and "off" conditions). RESULTS: Apathy decreased under both STN stimulation and levodopa treatment, whereas explicit and implicit stimulus reward learning was unchanged. CONCLUSIONS: Bilateral STN stimulation in PD patients does not necessarily have a negative effect on motivation and reward sensitivity and can even improve apathy provided patients have been appropriately selected for neurosurgery. PMID- 15897498 TI - Bilateral subthalamic nucleus stimulation improves balance control in Parkinson's disease. AB - BACKGROUND: Parkinson's disease (PD), the most common basal ganglia degenerative disease, affects balance control, especially when patients change balance strategy during postural tasks. Bilateral chronic stimulation of the subthalamic nucleus (STN) is therapeutically useful in advanced PD, and reduces the motor signs of patients. Nevertheless, the effects of STN stimulation on postural control are still debatable. AIMS: To assess the impact of bilateral STN stimulation on balance control in PD and to determine how basal ganglia related sensorimotor modifications act on neurosensorial organisation of balance and motor postural programming. METHODS: Twelve subjects aged 45-70 years underwent unified Parkinson's disease rating scale motor (part III) clinical tests, static and dynamic posturography, including sensory organisation and adaptation tests, shortly before and six months after bilateral implantation of electrodes into the STN. RESULTS: The postoperative static test showed an improvement in postural control precision both in eyes open and eyes closed conditions. The dynamic test highlighted the decreased number of falls and the ability of the patients to develop more appropriate sensorimotor strategies when stimulated. The sensory organisation test showed an improvement of equilibrium score and, thus, a better resolution of sensorial conflicts. CONCLUSIONS: STN stimulation allowed a reduction in rigidity and therefore an improvement in the ability to use muscular proprioception as reliable information, resulting in vestibulo-proprioceptive conflict suppression. STN stimulation has a synergistic effect with levodopa for postural control. Accordingly, non-dopaminergic pathways could be involved in postural regulation and STN stimulation may influence the functioning of these pathways. PMID- 15897499 TI - Neurodevelopmental treatment after stroke: a comparative study. AB - BACKGROUND: Neurodevelopmental treatment (NDT) is a rehabilitation approach increasingly used in the care of stroke patients, although no evidence has been provided for its efficacy. OBJECTIVE: To investigate the effects of NDT on the functional status and quality of life (QoL) of patients with stroke during one year after stroke onset. METHODS: 324 consecutive patients with stroke from 12 Dutch hospitals were included in a prospective, non-randomised, parallel group study. In the experimental group (n = 223), nurses and physiotherapists from six neurological wards used the NDT approach, while conventional treatment was used in six control wards (n = 101). Functional status was assessed by the Barthel index. Primary outcome was "poor outcome", defined as Barthel index <12 or death after one year. QoL was assessed with the 30 item version of the sickness impact profile (SA-SIP30) and the visual analogue scale. RESULTS: At 12 months, 59 patients (27%) in the NDT group and 24 (24%) in the non-NDT group had poor outcome (corresponding adjusted odds ratio = 1.7 (95% confidence interval, 0.8 to 3.5)). At discharge the adjusted odds ratio was 0.8 (0.4 to 1.5) and after six months it was 1.6 (0.8 to 3.2). Adjusted mean differences in the two QoL measures showed no significant differences between the study groups at six or 12 months after stroke onset. CONCLUSIONS: The NDT approach was not found effective in the care of stroke patients in the hospital setting. Health care professionals need to reconsider the use of this approach. PMID- 15897500 TI - Extent of white matter lesions is related to acute subcortical infarcts and predicts further stroke risk in patients with first ever ischaemic stroke. AB - OBJECTIVE: To investigate whether the extent of white matter lesions (WML) on fluid attenuated inversion recovery (FLAIR) MRI sequences is an independent risk factor for recurrent stroke, and to document the pattern of acute cerebral infarcts using diffusion weighted imaging (DWI) in patients with different severities of WML. METHODS: In a prospective cohort study, 228 consecutive stroke patients were studied between 1999 and 2001 in a community hospital. The severity of WML was graded as 0 (no WML), 1 (mild), 2 (moderate), or 3 (severe) according to the FLAIR appearances. DWI was used to document the location and size of the infarct. RESULTS: 31 patients had grade 0 WML, 69 had grade 1, 59 had grade 2, and 69 had grade 3. Age was independently associated with WML on logistic regression analysis (p = 0.0001). Acute cerebral infarcts in deep white matter were correlated with increasing severity of WML. On a median follow up of 23.0 months, life table analysis showed that recurrent stroke was related to the severity of WML (recurrence rate 7.8% in grade 0, 9.3% in grade 1, 17.7% in grade 2, 43.7% in grade 3; p = 0.0001). Survival was reduced in patients with severe WML (p = 0.0068). A Cox proportional hazards model showed WML to be predictive of recurrent stroke (p = 0.000, hazard ratio = 4.177 (95% confidence interval, 2.038 to 8.564)) and also for survival (p = 0.040, hazard ratio = 2.021 (1.032 to 3.960)). CONCLUSIONS: Patients with severe leukoaraiosis have increased risk of deep subcortical stroke and a higher risk of recurrent stroke. PMID- 15897501 TI - Changes in artistic style after minor posterior stroke. AB - BACKGROUND: Two professional painters experienced significant changes in their art as the main consequence of minor stroke located in the left occipital lobe or thalamus. METHODS: The features of this artistic conversion were analysed on the basis of extensive neurological, neuropsychological, and psychiatric evaluations. RESULTS: Both painters, initially unaware of the artistic changes, exhibited mild signs of executive dysfunction, but no general cognitive decline. The first painter, who showed mild visual-perceptive difficulties (dyschromatopsia and scotoma in his right upper visual field after left occipital stroke), together with increased anxiety and difficulty in emotional control, switched to a more stylised and symbolic art. The second painter, who also presented features of emotionalism related to his left latero-thalamic stroke, switched from an impressionist style to a more joyous and geometric, but more simplistic, abstract art. CONCLUSIONS: These findings show that mild cognitive and affective modifications due to focal posterior brain lesions can have significant repercussions on artistic expression. PMID- 15897502 TI - Positional and sleep dyspnoea due to posterior exophytic ependymoma of the medulla oblongata. PMID- 15897503 TI - Isolated monoparesis following stroke. AB - BACKGROUND: Some investigators have stated that monoparesis is almost never the result of a lacunar infarct or cerebral haemorrhage. OBJECTIVE: To describe the topography and aetiology in a consecutive population where first ever stroke was manifested by isolated monoparesis. METHODS: Patients with motor paresis of only one limb were included consecutively in the study. A neuroradiologist determined stroke location, while a neurologist reviewed the clinical records to assign stroke subtype. Both physicians worked blind to each other's findings. RESULTS: 51 of 2003 patients (2.5%) had isolated monoparesis, and of these 39 (76.5%) were ischaemic strokes and 12 (23.5%) were haemorrhagic. Cardioembolism was the cause of stroke in 15.7%, atherosclerosis in 9.8%, and small artery disease in 39.2%. Most of the haemorrhages were in the thalamic-capsular region (5/12). Most of the ischaemic lesions were in the deep territory of the middle cerebral artery, the corona radiate, or the centrum semiovale (20/39); 16 of 39 were in the cortical territories or the watershed region. CONCLUSIONS: Isolated monoparesis is a rare symptom in stroke patients and is often caused by small artery disease or a small haemorrhage. PMID- 15897504 TI - Interictal language functions in temporal lobe epilepsy. AB - OBJECTIVE: To evaluate interictal language functions in patients with medically intractable left and right sided mesial temporal lobe epilepsy (TLE). METHODS: Spontaneous speech, language comprehension, confrontation naming, repetition, reading, writing, and word fluency were examined in 12 patients with left sided TLE and 11 patients with right sided TLE. RESULTS: Four patients out of 23 displayed language deficits in more than one language domain. Three further patients exhibited isolated language deficits. Linguistic deficits were observed in both left TLE and right TLE. In quantitative analyses left and right TLE only differed in spontaneous speech (p = 0.02); no difference was found in other language functions, laterality quotient of Wada test, or overall IQ. Qualitative error analysis of object naming, however, showed typical errors associated only with left TLE. Patients with linguistic deficits were older at testing compared to patients without linguistic deficits (p = 0.003), whereas other factors including side of TLE, handedness, educational level, age at epilepsy onset, and duration of epilepsy did not differ between groups. CONCLUSIONS: Possible explanations for these findings include neuronal cell loss and deafferentiation in cortical areas, and disruption of the basal temporal language area pathways. Our study suggests that some patients with chronic mesial TLE exhibit linguistic deficits when specifically tested, and underlines the need to routinely investigate linguistic functions in TLE. PMID- 15897505 TI - Temporal lobe epilepsy surgery: different surgical strategies after a non invasive diagnostic protocol. AB - AIM: To test a non-invasive presurgical protocol for temporal lobe epilepsy (TLE) based on "anatomo-electro-clinical correlations". METHODS: All consecutive patients with suspected TLE and seizure history <2 years were entered into the protocol, which included video-electroencephalographic (EEG) monitoring and magnetic resonance imaging (MRI). Three different TLE subsyndromes (mesial, lateral, mesiolateral) were identified by combined anatomical, electrical, and clinical criteria. "Tailored" surgery for each subsyndrome was offered. Patients with seizure history <2 years, MRI evidence of temporal mass lesion, and concordant interictal EEG and clinical data bypassed video-EEG monitoring and were directly scheduled for surgery. RESULTS: Lesionectomy was performed without video-EEG recording in 11 patients with tumorous TLE. Of 146 patients studied with video-EEG, 133 received a TLE diagnosis. Four were excluded for neuropsychological risks, eight refused surgery, and 121 underwent surgery. Of 132 consecutive patients who underwent surgery, 101 had at least one year of follow up. They were divided into a "hippocampal sclerosis/cryptogenic" group (n = 57) and a "tumours/cortical organisation disorders" group (n = 44). In the first group, extensive temporal lobectomy (ETL) was performed in 40 patients, anteromesial temporal lobectomy (AMTL) in 17 patients. At follow up, 47 patients were seizure free. In the second group, lesionectomy plus ETL was performed in 23 patients, lesionectomy plus AMTL in six patients, and lesionectomy alone in 15 patients. Thirty nine patients were seizure free. CONCLUSIONS: These findings suggest that different TLE subsyndromes can be identified accurately using non invasive anatomo-electro-clinical data and can be treated effectively and safely with tailored surgery. PMID- 15897506 TI - Giant axonal neuropathy: clinical and genetic study in six cases. AB - BACKGROUND: Giant axonal neuropathy (GAN) is a severe recessive disorder characterised by variable combination of progressive sensory motor neuropathy, central nervous system (CNS) involvement, and "frizzly" hair. The disease is caused by GAN gene mutations on chromosome 16q24.1. AIMS: To search for GAN gene mutations in Turkish patients with GAN and characterise the phenotype associated with them. METHODS: Linkage and mutation analyses were performed in six affected patients from three consanguineous families. These patients were also investigated by cranial magnetic resonance imaging (MRI) and electroencephalography (EEG). Electromyography (EMG) was performed in heterozygous carriers from family 1 and family 3. RESULTS: Linkage to 16q24.1 was confirmed by haplotype analysis. GAN mutations were identified in all families. Family 1 had the R293X mutation, previously reported in another Turkish family. Families 2 and 3, originating from close geographical areas, shared a novel mutation, 1502+1G>T, at the donor splice site of exon 9. All patients displayed a common phenotype, including peripheral neuropathy, cerebellar ataxia, and frizzly hair. Cranial MRI showed diffuse white matter abnormalities in two patients from family 1 and the patient from family 3, and minimal white matter involvement in the patient from family 2. EMG of a heterozygous R293X mutation carrier showed signs of mild axonal neuropathy, whereas a 1502+1G>T mutation carrier had normal EMG. EEG abnormalities were found in three patients. CONCLUSION: These findings highlight the association of CNS involvement, in particular white matter abnormalities, with peripheral neuropathy in GAN. The phenotypical consequences of both mutations (when homozygous) were similar. PMID- 15897507 TI - Longlasting antalgic effects of daily sessions of repetitive transcranial magnetic stimulation in central and peripheral neuropathic pain. AB - BACKGROUND AND OBJECTIVE: A single session of repetitive transcranial magnetic stimulation (rTMS) over motor cortex had been reported to produce short term relief of some types of chronic pain. The present study investigated whether five consecutive days of rTMS would lead to longer lasting pain relief in unilateral chronic intractable neuropathic pain. PATIENTS AND METHODS: Forty eight patients with therapy resistant chronic unilateral pain syndromes (24 each with trigeminal neuralgia (TGN) and post-stroke pain syndrome (PSP)) participated. Fourteen from each group received 10 minutes real rTMS over the hand area of motor cortex (20 Hz, 10x10 s trains, intensity 80% of motor threshold) every day for five consecutive days. The remaining patients received sham stimulation. Pain was assessed using a visual analogue scale (VAS) and the Leeds assessment of neuropathic symptoms and signs (LANSS) scale, before, after the first, fourth, and fifth sessions, and two weeks after the last session. RESULTS: No significant differences were found in basal pain ratings between patients receiving real- and sham-rTMS. However, a two factor ANOVA revealed a significant "+/- TMS" x "time" interaction indicating that real and sham rTMS had different effects on the VAS and LANSS scales. Post hoc testing showed that in both groups of patients, real rTMS led to a greater improvement in scales than sham-rTMS, evident even two weeks after the end of the treatment. No patient experienced adverse effects. CONCLUSION: These results confirm that five daily sessions of rTMS over motor cortex can produce longlasting pain relief in patients with TGN or PSP. PMID- 15897508 TI - Applicability and relevance of models that predict short term outcome after intracerebral haemorrhage. AB - OBJECTIVES: Several models for prediction of short term outcome after intracerebral haemorrhage (ICH) have been published, however, these are rarely used in clinical practice for treatment decisions. This study was conducted to identify current models for prediction of short term outcome after ICH and to evaluate their clinical applicability and relevance in treatment decisions. METHODS: MEDLINE was searched from 1966 to June 2003 and studies were included if they met predefined criteria. Regression coefficients of multivariate models were extracted. Two neurologists independently evaluated the models for applicability in clinical practice. To assess clinical relevance and accuracy of each model, in a validation series of 122 patients the proportion with a >or=95% probability of death or poor outcome and the actual 30 day case fatality in these patients were calculated. Receiver operator characteristic (ROC) curves were computed for assessment of discriminatory power. RESULTS: A total of 18 prognostic models were identified, of which 14 appeared easy to apply. In the validation series, the proportion of patients with a >or=95% probability of death or poor outcome ranged from 0% to 43% (median 23%). The 30 day case fatality in these patients ranged from 75% to 100% (median 93%). The area under the ROC curves ranged from 0.81 to 0.90. CONCLUSIONS: Most models are easy to apply and can generate a high probability of death or poor outcome. However, only a small proportion of patients have such a high probability, and 30 day case fatality is not always correctly predicted. Therefore, current models have limited relevance in triage, but can be used to estimate the chances of survival of individual patients. PMID- 15897509 TI - Contribution of intraoperative electrical stimulations in surgery of low grade gliomas: a comparative study between two series without (1985-96) and with (1996 2003) functional mapping in the same institution. AB - OBJECTIVES: Despite the growing use of intraoperative functional mapping in supratentorial low grade glioma (LGG) surgery, few studies have compared series of patients operated on without and with direct electrical stimulation (DES) by the same team. The present study compared the rate of LGG surgery performed in eloquent areas, the rate of postoperative sequelae, and the quality of resection during two consecutive periods in the same department-the first without and the second with the use of intraoperative electrophysiology. METHODS: Between 1985 and 1996, 100 patients harbouring a supratentorial LGG underwent surgery with no functional mapping (S1). Between 1996 and 2003, 122 patients were operated on in the same department for a supratentorial LGG using intraoperative cortico subcortical DES (S2). RESULTS: Comparison between the two series showed that 35% of LGGs were operated on in eloquent areas in S1 versus 62% in S2 (p<0.0001), with 17% severe permanent deficits in S1 versus 6.5% in S2 (p<0.019). On postoperative MRI, 37% of resections were subtotal and 6% total in S1 versus 50.8% and 25.4%, respectively, in S2 (p<0.001). In both groups, survival was significantly related to the quality of resection. CONCLUSIONS: The results of the present study allow, for the first time, quantification of the contribution of intraoperative DES in LGG resection. Indeed, the use of this method leads to the extension of indications of LGG surgery within eloquent areas; to a decrease in the risk of sequelae; and to improvement of the quality of tumour resection, with an impact on survival. PMID- 15897510 TI - Brain lesions and eating disorders. AB - OBJECTIVE: To evaluate the relation between lesions of various brain structures and the development of eating disorders and thus inform the neurobiological research on the aetiology of these mental illnesses. METHOD: We systematically reviewed 54 previously published case reports of eating disorders with brain damage. Lesion location, presence of typical psychopathology, and evidence suggestive of causal association were recorded. RESULTS: Although simple changes in appetite and eating behaviour occur with hypothalamic and brain stem lesions, more complex syndromes, including characteristic psychopathology of eating disorders, are associated with right frontal and temporal lobe damage. CONCLUSIONS: These findings challenge the traditional view that eating disorders are linked to hypothalamic disturbance and suggest a major role of frontotemporal circuits with right hemispheric predominance in the pathogenesis. PMID- 15897511 TI - The John Hughlings Jackson 1935 Centenary Congress Medal. PMID- 15897512 TI - Genetic heterogeneity in rapid onset dystonia-parkinsonism: description of a new family. AB - Rapid onset dystonia-parkinsonism (RDP) is a rare movement disorder with autosomal dominant inheritance, characterised by sudden onset of dystonic spasms and slowness of movement. To date, three families have been described that share linkage to the same location on chromosome 19q13, designated DYT12. Very recently, mutations in the ATP1A3 gene at the DYT12 locus have been demonstrated in seven unrelated patients, including the three previously linked families. A large RDP family is reported here, with eight definitely and one possibly affected members, that is not linked to the DYT12 region and has no mutation in the ATP1A3 gene. Predominant cranial-cervical involvement of dystonia occurred in this family, which has also been described in patients with idiopathic torsion dystonia linked to the DYT6 region on chromosome 8 and is a rare finding in DYT1 dystonia. Molecular genetic analysis also excluded linkage to the DYT6 locus and the GAG deletion in DYT1, suggesting at least one additional RDP gene. PMID- 15897513 TI - A study of the workload and effectiveness of a comprehensive acute stroke service. AB - OBJECTIVE: To study the workload of and use of acute intervention within an established acute stroke service, the Calgary Stroke Programme (CSP). METHODS: Prospective record of all acute referrals, diagnoses, and management decisions over a 4 month period. RESULTS: The CSP received 572 referrals (median: 32 per week), 88% of which were made between 7 am and midnight. Of the 427 patients seen in person, 29% had not had an acute stroke or transient ischaemic attack (TIA). Fifty percent of patients with suspected acute stroke were referred within 3 h of symptom onset and 11% with acute ischaemic stroke (equating to 35% of those referred within 3 h of onset and seen in person) were treated with thrombolysis. CONCLUSION: Centralisation of services facilitates the rapid referral of, and use of acute interventions in, patients with acute stroke and TIA. Centralised services are likely to be busy (although less so at night), to attract large numbers of patients with disorders that mimic stroke and TIA, and yet still likely to treat only the minority of acute strokes using thrombolysis. These observations may help those planning similar services and underline the need to develop more widely applicable treatments for acute stroke. PMID- 15897514 TI - Prognosis of conservatively treated patients with Pott's paraplegia: logistic regression analysis. AB - OBJECTIVE: To evaluate the prognostic significance of various clinical, radiological, and neurophysiological findings in conservatively treated patients with Pott's paraplegia, using multiple regression analysis. METHODS: The study included 43 patients with Pott's paraplegia, managed conservatively. The diagnosis of Pott's spine was based on clinical, magnetic resonance imaging, and computed tomography or ultrasound guided aspiration biopsy. All patients were examined clinically, and motor evoked potentials (MEPs) to lower limbs and tibial somatosensory evoked potentials (SEP) were recorded. Outcome at six months was defined as good or poor. For evaluating predictors of outcome, 15 clinical, investigative, and evoked potential variables were analysed, using multiple logistic regression analysis. RESULTS: The age range of the patients was 16-70 years, and 22 were female. Mild spasticity with hyperreflexia only was seen in 13 patients. In the remaining, weakness was severe in eight, and moderate and mild in 11 patients each. Twenty patients had loss of joint position sensation. MEP and SEP were abnormal in 19 and 18 patients, respectively. On multiple regression analysis, the best model predicting six month outcome included power, paraplegia score, SEP, and MEP. CONCLUSION: Patients with Pott's paraplegia are likely to recover completely by six months if they have mild weakness, lower paraplegia score and normal SEPs and MEPs. PMID- 15897515 TI - Significance of Beevor's sign in facioscapulohumeral dystrophy and other neuromuscular diseases. AB - An atypical presentation of facioscapulohumeral dystrophy (FSH) is described, where the presence of a positive Beevor's sign led to genetic testing and subsequent probable diagnostic confirmation. This prompted evaluation of a further 68 patients for the presence of Beevor's sign. Among these, 19/20 patients with FSH had a positive Beevor's sign, compared with 2/28 with other muscle diseases, and 0/20 in a neurological control group. Beevor's sign should be considered as an additional criterion for the diagnosis of FSH. PMID- 15897516 TI - Mills' and other isolated upper motor neurone syndromes: in vivo study with 11C (R)-PK11195 PET. AB - (11)C-(R)-PK11195 positron emission tomography (PET) was used to explore and delineate in vivo the cortical lesion in three clinically isolated upper motor neurone syndromes of similar presentation, with reference to the syndrome of progressive spastic hemiparesis first described by Mills. Three patients with isolated UMN syndromes underwent (11)C-(R)-PK11195 PET of the brain. One patient fulfilled criteria for PLS. Two others had clinical features similar to the cases described by Mills; one of these had a high cervical cord inflammatory lesion previously noted on magnetic resonance imaging. The patient with PLS showed focal increase in the binding of (11)C-(R)-PK11195, indicating microglial activation, in the motor cortex contralateral to the predominantly affected limbs. Of the other two patients, one demonstrated marked increases in binding in the superior frontal region (supplementary motor region) contralateral to the affected limbs. In contrast, no focal areas of increased binding were seen in the cerebral cortex of the third patient, who had a high cervical cord lesion and was presumed to have extra-cerebral inflammatory disease. (11)C-(R)-PK11195 PET demonstrates in vivo that active pathology may be detectable many years after the onset of symptoms, and that it can occur in disparate sites with clinically similar presentations. We conclude that Mills' syndrome is a purely clinical description that should be reserved for patients with a progressive spastic hemiparesis for which no other explanation can be found. PMID- 15897517 TI - The electrophysiological profile of hereditary motor and sensory neuropathy-Lom. AB - OBJECTIVE: To make electrophysiological observations on a large kindred with hereditary motor and sensory neuropathy-Lom (HMSN-L) containing 27 affected individuals. CLINICAL FINDINGS: Onset was in early childhood with gait difficulty related to progressive lower limb weakness. Upper limb weakness developed later. Bulbar involvement was present in one third of the patients, and deafness appeared during the second or third decades. ELECTROPHYSIOLOGICAL FINDINGS: Electromyographic evidence of denervation was progressive, more severe distally, and greater in the legs, being total in distal lower limb muscles in most patients. Sensory action potentials were absent and motor nerve conduction was severely slowed. This included proximal upper limb (musculocutaneous and axillary), hypoglossal, and facial nerves. The severity of slowing increased during childhood. M waves, often multiple, were recorded in all affected individuals. The blink reflex showed an unusual three component response. The latencies of all three components were prolonged. CONCLUSIONS: HMSN-L is shown to be a demyelinating neuropathy involving severe and early axonal loss. The progressive slowing of nerve conduction during childhood differs from the static reduction seen in type I HMSN. PMID- 15897518 TI - Follow up of patients with signs and symptoms of polyneuropathy not confirmed by electrophysiological studies. AB - The outcome and final diagnoses of patients with symptoms and/or signs suggestive of polyneuropathy, but with normal electrophysiological studies, were investigated. All patients who presented at the outpatient clinic between 1993 and 1998 with signs and symptoms suggestive of polyneuropathy, but in whom electrophysiological studies were normal, were included. We retrospectively collected data from the medical records and then interviewed the patients and used the Sickness Impact Profile scale to investigate functional status at least 2 years after presentation. We included 74 patients, of whom 39 had neurological signs at neurological examination at the first visit. A final diagnosis was made in 24 of the 39 patients with neurological signs, and in three of the 35 patients without neurological signs but with symptoms. One (3%) of the 35 patients without neurological signs at the first visit had a poor outcome versus 15 (39%) of the 39 patients with neurological signs. In 11 (41%) of the 27 patients in the group with a final diagnosis the outcome was poor versus 5 (11%) of 47 patients without a final diagnosis. In 11 patients we concluded that they probably had small fibre neuropathy. Patients presenting with symptoms of polyneuropathy but who have neither neurological signs of polyneuropathy nor electrophysiological studies confirming a polyneuropathy have a good outcome at least 2 years after presentation. Further investigations are not indicated, except for patients fulfilling the criteria of small fibre neuropathy. In patients with neurological signs, as the outcome depends on the diagnosis and an explanation for these signs is often found, repeated investigations in this group are mandatory. PMID- 15897519 TI - The "pulvinar sign" in a case of paraneoplastic limbic encephalitis associated with non-Hodgkin's lymphoma. AB - This paper reports a 59 year old woman with paraneoplastic limbic encephalitis associated with diffuse large B cell lymphoma. Her brain magnetic resonance imaging scan showed bilateral posterior thalamic hyperintensities, similar to the "pulvinar sign". Her symptoms included progressive psychiatric disturbance and resembled the initial symptoms of variant Creutzfeldt-Jakob disease (vCJD). Clinicians should consider this treatable disorder in the differential diagnosis of vCJD. PMID- 15897520 TI - Ictal asystole with convulsive syncope mimicking secondary generalisation: a depth electrode study. AB - Ictal bradycardia is rare and its localising value is debated. Bradyarrhythmias are, however, important because of their potential connection to sudden death and ability to affect clinical seizure manifestations. Cerebral hypoperfusion induces loss of consciousness, at times with myoclonic jerks, whose clinical differentiation from a generalised convulsive seizure may prove difficult. Two invasive and five surface monitored seizures recorded over two years in a 51 year old woman with post-traumatic epilepsy characterised by seizure-triggered asystole were analysed. All seven seizures showed left temporal onset. Both intracranially recorded events started in the left anterior hippocampus/amygdala, spreading to the contralateral hippocampus in 35 and 25 seconds. Within 10 seconds an electrocardiogram showed asystole lasting 21 and 28 seconds, associated with suppression of recorded cerebral electrical activity, except a polyspike suppression pattern remaining in the hippocampi. Clinically, the patient, concomitantly with the cerebral suppression, developed myoclonic twitches of the limbs. A dual chamber cardiac pacemaker was implanted; at 11 months follow up, the patient has experienced only infrequent partial seizures, with none involving falls or shaking. Left temporal lobe seizures produced convulsive syncope initiated by ictal asystole. These observations suggest that intertemporal spread is necessary, though not sufficient, to produce bradycardia and asystole. Furthermore, pacemakers may decrease seizure severity, as well as potentially protect against malignant bradyarrhythmias. PMID- 15897521 TI - Recurrent hypertensive brainstem encephalopathy. PMID- 15897522 TI - Medial medullary infarction with contralateral glossoplegia. PMID- 15897523 TI - Diffuse cystic leucoencephalopathy after buprenorphine injection. PMID- 15897524 TI - Successful treatment of CMV ventriculitis immune reconstitution syndrome. PMID- 15897526 TI - Summaries for patients. Can a new antibiotic safely prevent travelers' diarrhea? PMID- 15897527 TI - Summaries for patients. Are high sugar levels associated with new cardiovascular events in postmenopausal women who have already had a heart attack? PMID- 15897528 TI - Summaries for patients. Antiviral regimens for chronic hepatitis B virus infection. PMID- 15897529 TI - Summaries for patients. Hormone therapy to prevent chronic conditions in postmenopausal women: recommendations from the U.S. Preventive Services Task Force. PMID- 15897530 TI - A randomized, double-blind, placebo-controlled trial of rifaximin to prevent travelers' diarrhea. AB - BACKGROUND: Travelers' diarrhea causes substantial morbidity and postinfectious irritable bowel syndrome. OBJECTIVE: To evaluate nonabsorbable rifaximin for prevention of travelers' diarrhea. DESIGN: Randomized, double-blind, placebo controlled clinical trial. SETTING: Guadalajara, Mexico. PARTICIPANTS: U.S. students. INTERVENTION: On arrival in Guadalajara, Mexico, 210 U.S. adults received rifaximin (200 mg/d, 200 mg twice daily, or 200 mg 3 times daily) or placebo for 2 weeks. MEASUREMENTS: Participants were followed daily for 3 weeks for enteric disease and symptoms and daily for 5 weeks for drug side effects. Changes in intestinal coliform flora were studied. RESULTS: Travelers' diarrhea developed in 14.74% of participants taking rifaximin and 53.70% of those taking placebo (rate ratio, 0.27 [95% CI, 0.17 to 0.43]). Rifaximin provided 72% and 77% protection against travelers' diarrhea and antibiotic-treated travelers' diarrhea, respectively (P < 0.001 for both), and all rifaximin doses were superior to placebo. In the groups that did not report travelers' diarrhea, rifaximin significantly reduced the occurrence of mild diarrhea (P = 0.02) and moderate and severe intestinal problems (P = 0.009 for pain or cramps; P = 0.02 for excessive gas). Rates of adverse events were comparable in the rifaximin and placebo groups. Minimal changes in coliform flora were found during rifaximin therapy. LIMITATIONS: Rifaximin safely prevented travelers' diarrhea in Mexico, where most cases are caused by diarrhea-producing Escherichia coli. A study is needed in Asia to determine whether rifaximin can prevent diarrhea caused by invasive bacterial pathogens. CONCLUSIONS: Rifaximin prevents travelers' diarrhea with minimal changes in fecal flora, and more liberal chemoprophylaxis against this disease should be considered. Future studies should evaluate whether rifaximin is effective in preventing postinfectious irritable bowel syndrome. PMID- 15897531 TI - Impaired fasting glucose and cardiovascular outcomes in postmenopausal women with coronary artery disease. AB - BACKGROUND: Type 2 diabetes increases risk for cardiovascular disease. Persons with impaired fasting glucose levels may also have increased risk. OBJECTIVE: To evaluate the association between glucose status and cardiovascular outcomes and the effect of lowering the fasting glucose level criterion for impaired fasting glucose from a lower limit of 6.1 mmol/L (110 mg/dL) to 5.6 mmol/L (100 mg/dL). DESIGN: Prospective cohort study. SETTING: 20 U.S. clinical centers. PATIENTS: 2763 postmenopausal women with established coronary heart disease (CHD) who were followed for 6.8 years. MEASUREMENTS: Any CHD event (nonfatal myocardial infarction or CHD death), stroke or transient ischemic attack (TIA), congestive heart failure (CHF) hospitalization, and any cardiovascular event. RESULTS: During follow-up, 583 women had a CHD event, 329 women had a stroke or TIA, and 348 women were hospitalized for CHF. Women with diabetes were at an approximately 75% increased risk for each outcome compared with normoglycemic women. The 218 women with impaired fasting glucose according to the 1997 definition (fasting glucose level, 6.1 to 6.9 mmol/L [110 to 125 mg/dL]) had increased risk for any CHD event (hazard ratio, 1.37 [95% CI, 1.08 to 1.74]), while the 698 women with impaired fasting glucose according to the 2003 definition (fasting glucose level, 5.6 to 6.9 mmol/L [100 to 125 mg/dL]) were not at increased risk (hazard ratio, 1.09 [CI, 0.90 to 1.34]). Most of the women (n = 480) with fasting glucose levels between 5.6 mmol/L (100 mg/dL) and 6.0 mmol/L (109 mg/dL) had no increased risk for CHD (hazard ratio, 0.90 [CI, 0.73 to 1.12]). Women with impaired fasting glucose according to either definition were not at increased risk for stroke or TIA or CHF. LIMITATIONS: These findings may not be generalizable to men or women without existing heart disease. CONCLUSIONS: Among postmenopausal women with coronary artery disease, the 2003 definition for impaired fasting glucose was not associated with increased risk for new CHD, stroke or TIA, or CHF. PMID- 15897532 TI - Treatment alternatives for chronic hepatitis B virus infection: a cost effectiveness analysis. AB - BACKGROUND: Treatment options for chronic hepatitis B virus (HBV) infection have disparate risks and benefits. Interferon has clinically significant side effects, and lamivudine is associated with viral resistance. In contrast, adefovir is safe and has lower viral resistance but is more expensive. The most cost-effective approach is uncertain. OBJECTIVE: To determine whether and under what circumstances the improved efficacy of adefovir offsets its increased cost compared with lamivudine or interferon. DESIGN: Cost-utility analysis stratified by hepatitis B e antigen (HBeAg) status. DATA SOURCES: Systematic review of MEDLINE from 1970 to 2005. TARGET POPULATION: Patients with chronic HBV infection, elevated aminotransferase levels, and no cirrhosis. TIME HORIZON: Lifetime. PERSPECTIVE: Third-party payer. INTERVENTIONS: 1) No HBV treatment ("do nothing" strategy), 2) interferon monotherapy, 3) lamivudine monotherapy, 4) adefovir monotherapy, or 5) lamivudine with crossover to adefovir upon resistance ("adefovir salvage" strategy). OUTCOME MEASURE: Incremental cost per quality adjusted life-year (QALY) gained. RESULTS OF BASE-CASE ANALYSIS: The "do nothing" strategy was least effective yet least expensive. Compared with the "do nothing" strategy, using interferon cost an incremental 6337 dollars to gain 1 additional QALY. Compared with interferon, the adefovir salvage strategy cost an incremental 8446 dollars per QALY gained. Both the lamivudine and adefovir monotherapy strategies were more expensive yet less effective than the alternative strategies and were therefore dominated. RESULTS OF SENSITIVITY ANALYSIS: In sensitivity analysis, interferon was most cost-effective in health care systems with tight budgetary constraints and a high prevalence of HBeAg-negative patients. LIMITATIONS: These results apply only to patients with chronic HBV infection, elevated aminotransferase levels, and no clinical or histologic evidence of cirrhosis. They do not apply to alternative populations. CONCLUSIONS: Neither lamivudine nor adefovir monotherapy is cost-effective in chronic HBV infection. However, a hybrid salvage strategy reserving adefovir only for lamivudine associated viral resistance may be highly cost-effective across most health care settings. Interferon therapy may still be preferred in health care systems with limited resources, especially in those serving populations with a high prevalence of HBeAg-negative HBV. PMID- 15897533 TI - Adults are whooping, but are internists listening? AB - Although studies have shown that pertussis is responsible for approximately 20% to 30% of cases of cough lasting more than 2 weeks in adults and adolescents, physicians have generally considered pertussis a disease that affects only children. Epidemiologic trends demonstrate that the incidence of pertussis has been rising, especially in adolescents and adults. Consequently, pertussis is not a "zebra" diagnosis but deserves a place among the community-acquired list of adult pathogens. Increased awareness among physicians about pertussis is important because adults may transmit the infection to infants, in whom the disease often leads to hospitalization and may result in death. Reporting this disease to the local health department is critical to its control because both case-patients and their close contacts should receive antimicrobial prophylaxis. A booster vaccine for adolescents and adults is likely to become available in the near future, so this is an important time to ensure that all physicians, not just pediatricians, are knowledgeable about the diagnosis, treatment, and reporting requirements for pertussis. PMID- 15897534 TI - Meta-analysis: accuracy of rapid tests for malaria in travelers returning from endemic areas. AB - BACKGROUND: Microscopic diagnosis of malaria is unreliable outside specialized centers. Rapid tests have become available in recent years, but their accuracy has not been assessed systematically. PURPOSE: To determine the accuracy of rapid diagnostic tests for ruling out malaria in nonimmune travelers returning from malaria-endemic areas. DATA SOURCES: The authors searched MEDLINE, EMBASE, CAB Health, and CINAHL (1988 to September 2004); hand-searched conference proceedings; checked reference lists; and contacted experts and manufacturers. STUDY SELECTION: Diagnostic accuracy studies in nonimmune individuals with suspected malaria were included if they compared rapid tests with expert microscopic examination or polymerase chain reaction tests. DATA EXTRACTION: Data on study and patient characteristics and results were extracted in duplicate. The main outcome was the likelihood ratio for a negative test result (negative likelihood ratio) for Plasmodium falciparum malaria. Likelihood ratios were combined by using random-effects meta-analysis, stratified by the antigen targeted (histidine-rich protein-2 [HRP-2] or parasite lactate dehydrogenase [LDH]) and by test generation. Nomograms of post-test probabilities were constructed. DATA SYNTHESIS: The authors included 21 studies and 5747 individuals. For P. falciparum, HRP-2-based tests were more accurate than parasite LDH-based tests: Negative likelihood ratios were 0.08 and 0.13, respectively (P = 0.019 for difference). Three-band HRP-2 tests had similar negative likelihood ratios but higher positive likelihood ratios compared with 2 band tests (34.7 vs. 98.5; P = 0.003). For P. vivax, negative likelihood ratios tended to be closer to 1.0 for HRP-2-based tests than for parasite LDH-based tests (0.24 vs. 0.13; P = 0.22), but analyses were based on a few heterogeneous studies. Negative likelihood ratios for the diagnosis of P. malariae or P. ovale were close to 1.0 for both types of tests. In febrile travelers returning from sub-Saharan Africa, the typical probability of P. falciparum malaria is estimated at 1.1% (95% CI, 0.6% to 1.9%) after a negative 3-band HRP-2 test result and 97% (CI, 92% to 99%) after a positive test result. LIMITATIONS: Few studies evaluated 3-band HRP-2 tests. The evidence is also limited for species other than P. falciparum because of the few available studies and their more heterogeneous results. Further studies are needed to determine whether the use of rapid diagnostic tests improves outcomes in returning travelers with suspected malaria. CONCLUSIONS: Rapid malaria tests may be a useful diagnostic adjunct to microscopy in centers without major expertise in tropical medicine. Initial decisions on treatment initiation and choice of antimalarial drugs can be based on travel history and post-test probabilities after rapid testing. Expert microscopy is still required for species identification and confirmation. PMID- 15897535 TI - High and rising health care costs. Part 1: seeking an explanation. AB - The United States has the most expensive health care system in the world, with per capita health expenditures far above those of any other nation. For many years, U.S. health care expenditures have been growing above the overall rate of inflation in the economy. A few experts have argued that high and rising costs are not such a serious problem. Most observers disagree with this view, pointing to the negative impact of employee health care costs on employers, the government budgetary problems caused by rising health care expenditures, and an association between high health care costs and reduced access for individuals needing health services. Several explanations have been offered for high and rising health care costs. These include the perspectives that high and rising costs are created by forces external to the health system, by the weakness of a competitive free market within the health system, by the rapid diffusion of new technologies, by excessive costs of administering the health system, by the absence of strong cost containment measures, and by undue market power of health care providers. This article, the first in a 4-part series, discusses 3 perspectives on health care: 1) Are high and rising health care costs a serious problem? 2) Are rising costs explained by factors outside the health care system? 3) Does the absence of a free market in health care explain why costs are high and rising? The remaining 3 articles in this series address other perspectives on health care costs. PMID- 15897536 TI - Hormone therapy for the prevention of chronic conditions in postmenopausal women: recommendations from the U.S. Preventive Services Task Force. AB - This statement summarizes the U.S. Preventive Services Task Force recommendations on hormone therapy for the prevention of chronic conditions in postmenopausal women and the supporting scientific evidence, and updates the Task Force's 2002 recommendations on hormone replacement therapy. The updated statement is based on the results of the Women's Health Initiative randomized, controlled trial, as well as the information in the 2002 summary of the evidence on this topic, which is available on the USPSTF Web site (http://www.preventiveservices.ahrq.gov). PMID- 15897537 TI - How to hit the runs for fifty million travelers at risk. PMID- 15897538 TI - Assessing the benefits and costs of new therapies for hepatitis B virus infection. PMID- 15897539 TI - Understanding rising health care costs: introducing a series of articles. PMID- 15897540 TI - Minnie made me out a liar. PMID- 15897541 TI - Acupuncture and knee osteoarthritis. PMID- 15897542 TI - The dry pipeline of antiarrhythmic therapies. PMID- 15897543 TI - Acupuncture and knee osteoarthritis. PMID- 15897544 TI - Acupuncture for chronic mechanical neck pain. PMID- 15897545 TI - Treatments for back pain. PMID- 15897546 TI - Long-term reduction of vascular access-associated bloodstream infection. PMID- 15897547 TI - Mainstream and alternative medicine. PMID- 15897548 TI - Is sleep the new vital sign? PMID- 15897549 TI - Epigenetic and chromatin modifiers as targeted therapy of hematologic malignancies. AB - Epigenetic regulation of gene expression is mediated through alterations in the DNA methylation status, covalent modifications of core nucleosomal histones, rearrangement of histones, and by RNA interference. It is now abundantly clear that deregulation of epigenetic mechanisms cooperates with genetic alterations in the development and progression of cancer and leukemia. Epigenetic deregulation affects several aspects of tumor cell biology, including cell growth, cell cycle control, differentiation, DNA repair, and cell death. This raises the strong possibility that reversing deregulated epigenetic mechanisms may be an effective treatment strategy for leukemia and cancer. This treatment strategy may either be designed to separately or collectively target the specific perturbations in the epigenetic mechanisms found in human hematologic malignancies. The following review describes our current understanding of the important deregulated epigenetic mechanisms and the preclinical and clinical development of epigenetic and chromatin modifiers in the therapy of these disorders. PMID- 15897550 TI - Phase I study of an oral histone deacetylase inhibitor, suberoylanilide hydroxamic acid, in patients with advanced cancer. AB - PURPOSE: To determine the safety, dosing schedules, pharmacokinetic profile, and biologic effect of orally administered histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) in patients with advanced cancer. PATIENTS AND METHODS: Patients with solid and hematologic malignancies were treated with oral SAHA administered once or twice a day on a continuous basis or twice daily for 3 consecutive days per week. Pharmacokinetic profile and bioavailibity of oral SAHA were determined. Western blots and enzyme-linked immunosorbent assays of histones isolated from peripheral-blood mononuclear cells (PBMNCs) pre and post-therapy were performed to evaluate target inhibition. RESULTS: Seventy-three patients were treated with oral SAHA and major dose-limiting toxicities were anorexia, dehydration, diarrhea, and fatigue. The maximum tolerated dose was 400 mg qd and 200 mg bid for continuous daily dosing and 300 mg bid for 3 consecutive days per week dosing. Oral SAHA had linear pharmacokinetics from 200 to 600 mg, with an apparent half-life ranging from 91 to 127 minutes and 43% oral bioavailability. Histones isolated from PBMNCs showed consistent accumulation of acetylated histones post-therapy, and enzyme-linked immunosorbent assay demonstrated a trend towards a dose-dependent accumulation of acetylated histones from 200 to 600 mg of oral SAHA. There was one complete response, three partial responses, two unconfirmed partial responses, and 22 (30%) patients remained on study for 4 to 37+ months. CONCLUSIONS: Oral SAHA has linear pharmacokinetics and good bioavailability, inhibits histone deacetylase activity in PBMNCs, can be safely administered chronically, and has a broad range of antitumor activity. PMID- 15897551 TI - Epigenetic profiling of cutaneous T-cell lymphoma: promoter hypermethylation of multiple tumor suppressor genes including BCL7a, PTPRG, and p73. AB - PURPOSE: To analyze the occurrence of promoter hypermethylation in primary cutaneous T-cell lymphoma (CTCL) on a genome-wide scale, focusing on epigenetic alterations with pathogenetic significance. MATERIALS AND METHODS: DNA isolated from biopsy specimens of 28 patients with CTCL, including aggressive CTCL entities (transformed mycosis fungoides and CD30-negative large T-cell lymphoma) and an indolent entity (CD30-positive large T-cell lymphoma), were investigated. For genome-wide DNA methylation screening, differential methylation hybridization using CpG island microarrays was applied, which allows simultaneous detection of the methylation status of 8640 CpG islands. Bisulfite sequence analysis was applied for confirmation and detection of hypermethylation of eight selected tumor suppressor genes. RESULTS: The DNA methylation patterns of CTCLs emerging from differential methylation hybridization analysis included 35 CpG islands hypermethylated in at least four of the 28 studied CTCL samples when compared with benign T-cell samples. Hypermethylation of the putative tumor suppressor genes BCL7a (in 48% of CTCL samples), PTPRG (27%), and thrombospondin 4 (52%) was confirmed and demonstrated to be associated with transcriptional downregulation. BCL7a was hypermethylated at a higher frequency in aggressive (64%) than in indolent (14%) CTCL entities. In addition, the promoters of the selected tumor suppressor genes p73 (48%), p16 (33%), CHFR (19%), p15 (10%), and TMS1 (10%) were hypermethylated in CTCL. CONCLUSION: Malignant T cells of patients with CTCL display widespread promoter hypermethylation associated with inactivation of several tumor suppressor genes involved in DNA repair, cell cycle, and apoptosis signaling pathways. In view of this, CTCL may be amenable to treatment with demethylating agents. PMID- 15897552 TI - Paclitaxel after doxorubicin plus cyclophosphamide as adjuvant chemotherapy for node-positive breast cancer: results from NSABP B-28. AB - PURPOSE: The primary aim of National Surgical Adjuvant Breast and Bowel Project (NSABP) B-28 was to determine whether four cycles of adjuvant paclitaxel (PTX) after four cycles of adjuvant doxorubicin/cyclophosphamide (AC) will prolong disease-free survival (DFS) and overall survival (OS) compared with four cycles of AC alone in patients with resected operable breast cancer and histologically positive axillary nodes. PATIENTS AND METHODS: Between August 1995 and May 1998, 3,060 patients were randomly assigned (AC, 1,529; AC followed by PTX [AC --> PTX], 1,531). Patients > or = 50 years and those younger than 50 years with estrogen receptor (ER) or progesterone receptor (PR) -positive tumors also received tamoxifen for 5 years, starting with the first dose of AC. Postlumpectomy radiotherapy was mandated. Postmastectomy or regional radiotherapy was prohibited. Median follow-up is 64.6 months. RESULTS: The addition of PTX to AC significantly reduced the hazard for DFS event by 17% (relative risk [RR], 0.83; 95% CI, 0.72 to 0.95; P = .006). Five-year DFS was 76% +/- 2% for patients randomly assigned to AC --> PTX compared with 72% +/- 2% for those randomly assigned to AC. Improvement in OS was small and not statistically significant (RR, 0.93; 95% CI, 0.78 to 1.12; P = .46). Five-year OS was 85% +/- 2% for both groups. Subset analysis of the effect of paclitaxel according to hormone receptors or tamoxifen administration did not reveal statistically significant interaction (for DFS, P = .30 and P = .44, respectively). Toxicity with the AC - > PTX regimen was acceptable for the adjuvant setting. CONCLUSION: The addition of PTX to AC resulted in significant improvement in DFS but no significant improvement in OS with acceptable toxicity. No significant interaction between treatment effect and receptor status or tamoxifen administration was observed. PMID- 15897553 TI - The pathway to biomarker discovery: carbonic anhydrase IX and the prediction of immune responsiveness. PMID- 15897554 TI - Approval summary: azacitidine for treatment of myelodysplastic syndrome subtypes. AB - PURPOSE: This article summarizes data submitted to the U.S. Food and Drug Administration for marketing approval of azacitidine as injectable suspension (Vidaza, Pharmion Corporation, Boulder, CO) for treatment of patients with myelodysplastic syndrome. EXPERIMENTAL DESIGN: In one phase 3 controlled trial, 191 study subjects were randomized to treatment with azacitidine or to observation; an additional 120 patients were treated with azacitidine in two phase 2 single arm studies. The primary efficacy end point was the overall response rate, defined as complete or partial normalization of peripheral blood counts and bone marrow blast percentages for at least 4 weeks. RESULTS: In the controlled trial, the overall response rate was 15.7% in the azacitidine treatment group; there were no responders in the observation group (P < 0.0001). Response rates were similar in the two single arm studies. During response patients stopped being red cell or platelet transfusion dependent. Median duration of responses was at least 9 months. An additional 19% of azacitidine treated patients had less than partial responses, most becoming transfusion independent. The most common adverse events attributed to azacitidine were gastrointestinal, hematologic, local (injection site), and constitutional. There were no azacitidine-related deaths. CONCLUSIONS: On May 19, 2004 the U.S. Food and Drug Administration approved azacitidine as injectable suspension for treatment of patients with the following myelodysplastic syndrome subtypes: refractory anemia or refractory anemia with ringed sideroblasts (if accompanied by neutropenia or thrombocytopenia or requiring transfusions), refractory anemia with excess blasts, refractory anemia with excess blasts in transformation, and chronic myelomonocytic leukemia. Full prescribing information is available at http://www.fda.gov/cder/foi/label/2004/050794lbl.pdf. Azacitidine is the first agent approved for treatment of myelodysplastic syndrome. PMID- 15897555 TI - DDEF1 is located in an amplified region of chromosome 8q and is overexpressed in uveal melanoma. AB - PURPOSE: The molecular pathogenesis of uveal melanoma is poorly understood but is usually accompanied by amplification of chromosome 8q, suggesting the activation of one or more oncogenes. We recently identified a gene expression profile that distinguishes low-grade from high-grade melanomas. In this profile, a cluster of genes at chromosome 8q was overexpressed in high-grade tumors, providing an opportunity to search for potential oncogenes in this region. EXPERIMENTAL DESIGN: Gene expression microarray analysis was done on 25 primary uveal melanomas. Microarray comparative genomic hybridization (CGH), quantitative PCR, and immunohistochemistry were done on a subset of these tumors. Cell motility was measured using a wound-healing assay. RESULTS: In melanomas analyzed for microarray gene expression and CGH, gain of chromosome 8q correlated most strongly with expression of DDEF1, a gene located at 8q24. In contrast, the nearby MYC oncogene exhibited no significant change in expression. Confirming the microarray findings, DDEF1 mRNA levels and protein expression were significantly higher in high-grade melanomas. Furthermore, ectopic expression of DDEF1 in low grade melanoma cells resulted in a significant increase in cell motility, a feature of high-grade metastasizing cells. CONCLUSIONS: These findings suggest that DDEF1 overexpression may be a pathogenetically relevant consequence of chromosome 8q amplification, which commonly occurs in high-grade uveal melanomas. We conclude that DDEF1 may act as an oncogene in this cancer, and it may be a useful diagnostic marker and therapeutic target. PMID- 15897556 TI - A rare transporter associated with antigen processing polymorphism overpresented in HLAlow colon cancer reveals the functional significance of the signature domain in antigen processing. AB - Transporter associated with antigen processing (TAP), a member of the ATP-binding cassette transporter superfamily, is composed of two integral membrane proteins, TAP-1 and TAP-2. Each subunit has a C-terminal nucleotide-binding domain that binds and hydrolyzes ATP to energize peptide translocation across the endoplasmic reticulum membrane. A motif comprising the sequence LSGGQ (called the signature motif) and the amino acid that is immediately C-terminal to this motif are highly conserved in the nucleotide-binding domains of ATP-binding cassette transporters. To search for natural variants of TAP-1 with alterations in or near the signature motif, we sequenced the TAP-1 exon 10 amplified from 103 human colon cancer samples. We found a rare TAP-1 allele with an R>Q alteration at a residue immediately C-terminal to the signature motif (R648) that occurred 17.5 times more frequently in colon cancers with down-regulated surface class I MHC than those with normal MHC levels (P = 0.01). Functional analysis revealed that the Q648 variant had significantly reduced peptide translocation activity compared with TAP-1 (R648). In addition, we found that mutations S644R, G645R, G646S, and G646D interfered with TAP-1 activity. TAP-1 G646D, which showed the most severe defect, resided normally in the endoplasmic reticulum and associated with the peptide loading complex, but failed to transport peptide across the endoplasmic reticulum membrane. Thus, a TAP-1 polymorphism adjacent to the signature motif may be a contributing factor for MHC class I down-regulation in colon cancer. Given the widespread defects in DNA mismatch repair in colon cancer, mutations at or near the signature domain can potentially modulate antigen processing. PMID- 15897557 TI - Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme. AB - INTRODUCTION: Research into the pathogenesis, molecular signaling, and treatment of glioblastoma multiforme (GBM) has traditionally been conducted using cell lines derived from malignant gliomas. We compared protein expression patterns between solid primary GBMs and GBM cell lines to identify proteins whose expression may be altered in cell culture. METHODS: We cultured cell lines U87, U118, U251, and A172 and used tissue-selective microdissection of eight primary GBMs to obtain pure populations of tumor cells, which we studied using two dimensional gel electrophoresis (2DGE) and examined using differential expression software. Select protein targets expressed differentially between GBM tumors and GBM cell lines were sequenced using tandem mass spectrometry. RESULTS: Analysis of the primary GBM tumor samples (n = 8) and the GBM cell lines revealed reproducibly similar proteomic patterns for each group, which distinguished tumors from the cell lines. Gels contained up to 500 proteins that were consistently identified in the pH 4 to 7 range. Comparison of proteins identified in the GBM tumors and in the cell lines showed approximately 160 proteins that were gained and 60 proteins that were lost on culture. Using normalized intensity patterns from the 2DGE images, ANOVA tests were done and statistically significant spots were identified. Seven proteins found in the cell lines were significantly increased when compared with the GBM tumors (P < 0.05), whereas 10 proteins were significantly decreased from cell lines compared with the GBM tumors. Proteins identified included transcription factors, tumor suppressor genes, cytoskeletal proteins, and cellular metabolic proteins. CONCLUSION: Global protein and proteomic differences were identified between primary GBM tumor samples and GBM cell lines. The proteins identified by 2DGE analysis elucidate some of the selection pressures of in vitro culture, help accentuate the advantages and limitations of cell culture, and may aid comprehension of gliomagenesis and enhance development of new therapeutics. PMID- 15897558 TI - In vivo target modulation and biological activity of CHIR-258, a multitargeted growth factor receptor kinase inhibitor, in colon cancer models. AB - PURPOSE: To evaluate the therapeutic and biological effects of CHIR-258, an orally bioavailable, potent inhibitor of class III-V receptor tyrosine kinases, in colon cancer models. EXPERIMENTAL DESIGN: The pharmacologic activity of CHIR 258 was characterized by monitoring target modulation as well as by evaluating the antitumor and antiangiogenic effects in human colon xenograft models. RESULTS: CHIR-258 inhibits vascular endothelial growth factor receptor 1/2, fibroblast growth factor receptor 1/3, and platelet-derived growth factor receptor beta (PDGFRbeta) and shows both antitumor and antiangiogenic activities in vivo. Treatment of KM12L4a human colon cancer cells with CHIR-258 resulted in a dose-dependent inhibition of vascular endothelial growth factor receptor 1 and PDGFRbeta phosphorylation and reduction of phosphorylated extracellular signal regulated kinase (ERK) levels, indicating modulation of target receptors and downstream signaling. In vivo administration of CHIR-258 resulted in significant tumor growth inhibition and tumor regressions, including large, established tumors (500-1,000 mm(3)). Immunohistochemical analysis showed a reduction of phosphorylated PDGFRbeta and phosphorylated ERK in tumor cells after oral dosing with CHIR-258 compared with control tumors. These changes were accompanied by decreased tumor cell proliferation rate and reduced intratumoral microvessel density. CHIR-258 inhibited the phosphorylation of PDGFRbeta and ERK phosphorylation in tumors within 2 hours following dosing and the inhibitory activity was sustained for >24 hours. Significant antitumor activity was observed with intermittent dosing schedules, indicating a sustained biological activity. CONCLUSION: These studies provide evidence that biological activity of CHIR-258 in tumors correlates with efficacy and aids in the identification of potential biomarkers of this multitargeted receptor tyrosine kinase inhibitor. CHIR-258 exhibits properties that make it a promising candidate for clinical development in a variety of solid and hematologic malignancies. PMID- 15897559 TI - Association of visceral fat accumulation and plasma adiponectin with colorectal adenoma: evidence for participation of insulin resistance. AB - PURPOSE: Colorectal carcinogenesis is thought to be related to abdominal obesity and insulin resistance. To investigate whether visceral fat accumulation contributes to colorectal carcinogenesis, we examined its accumulation and the levels of the adipose tissue-derived hormone adiponectin in Japanese patients with colorectal adenoma. EXPERIMENTAL DESIGN: Fifty-one consecutive Japanese patients ages >/=40 years and with colorectal adenoma were subjected to measurement of visceral fat area by computed tomography scanning and plasma adiponectin concentration. The patients also underwent the 75-g oral glucose tolerance test. Insulin resistance was calculated by the homeostasis metabolic assessment (HOMA-IR) method. The controls were 52 Japanese subjects ages >/=40 years and without colorectal polyp. Cigarette smokers and subjects who consumed alcohol (>/=30 g ethanol/d) were excluded. RESULTS: The patients with colorectal adenoma showed significantly more visceral fat area and significantly less plasma adiponectin concentration in comparison with the controls [odds ratio (OR), 2.19; 95% confidence interval (95% CI), 1.47-3.28; P < 0.001 and OR, 0.24; 95% CI, 0.14 0.41; P < 0.001, respectively] by logistic regression analysis. HOMA-IR index was also associated with colorectal adenoma (OR 2.60; 95% CI, 1.20-5.64; P = 0.040). Visceral fat area and adiponectin were associated with adenoma number (1, 2, >/= 3), the size of the largest adenoma (<10 and >/=10 mm), and adenoma histology (tubular and tubulovillous/villous). CONCLUSIONS: These results suggest an association of visceral fat accumulation and decreased plasma adiponectin concentration with colorectal adenoma in Japanese patients. This study may offer a new insight to understanding the relationship of colorectal carcinogenesis with abdominal obesity and insulin resistance. PMID- 15897560 TI - Vascular endothelial growth factor polymorphisms in relation to breast cancer development and prognosis. AB - PURPOSE: Angiogenesis is a necessary step in tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is a major mediator of breast cancer angiogenesis. Therefore, we investigated the association of polymorphisms in the VEGF gene with breast cancer risk and prognostic characteristics of the tumors in a large case-control study. EXPERIMENTAL DESIGN: We examined three polymorphisms in the VEGF gene (-2578C/A, -1154G/A, and +936C/T) in 571 familial breast cancer cases from Poland and Germany and -2578C/A, -634G/C, and +936C/T polymorphisms in 974 unselected breast cancer cases from Sweden together with ethnically and geographically selected controls. RESULTS: None of the polymorphisms or any haplotype was significantly associated with either familial or unselected breast cancers. Our study suggests that the +936C/T polymorphism is unlikely to be associated with breast cancer. We also analyzed the unselected cases for genotypes or haplotypes that associated with tumor characteristics. The -634CC genotype and the -2578/-634 CC haplotype were significantly associated with high tumor aggressiveness (large tumor size and high histologic grade, P < 0.01) and the -2578AA genotype and the -2578/-634 AG haplotype with low histologic grade tumors (P = 0.04). The genotypes and haplotypes were not related with other tumor characteristics such as regional or distant metastasis, stage at diagnosis, or estrogen or progesterone receptor status. CONCLUSIONS: Although none of the polymorphisms studied in the VEGF gene was found to influence susceptibility to breast cancer significantly, some of the VEGF genotypes and haplotypes may influence tumor growth through an altered expression of VEGF and tumor angiogenesis. PMID- 15897561 TI - Distinct methylation patterns of benign and malignant liver tumors revealed by quantitative methylation profiling. AB - PURPOSE: A comparative quantitative methylation profiling of hepatocellular carcinoma and the most frequent benign liver tumor, hepatocellular adenoma, was set up for the identification of tumor-specific methylation patterns. EXPERIMENTAL DESIGN: The quantitative methylation levels of nine genes (RASSF1A, cyclinD2, p16INK4a, DAP-K, APC, RIZ-1, HIN-1, GSTpi1, SOCS-1) were analyzed in hepatocellular carcinoma and adjacent normal tissue (n = 41), hepatocellular adenoma and adjacent normal tissue (n = 26), focal nodular hyperplasia (n = 10), and unrelated normal liver tissue (n = 28). Accumulated methylation data were analyzed using various statistical algorithms, including hierarchical clustering, to detect tumor-specific methylation patterns. RESULTS: Cluster analysis revealed that hepatocellular adenoma displays a methylation profile much more similar to that found in normal liver tissue and focal nodular hyperplasia than to that found in hepatocellular carcinoma. Many characteristic differences were not detected when using mere qualitative methylation assays. The cyclinD2 gene was identified as a new and frequent target for aberrant hypermethylation in hepatocellular carcinoma (68%). In the control group of 28 liver specimens from healthy donors, a clear correlation between age of patient and frequency and level of aberrant methylation was seen, which could not be detected in the group of hepatocellular carcinoma specimens. CONCLUSIONS: Methylation profiling can clearly contribute to the unequivocal classification of suspicious lesions, but only if done in a quantitative manner applying cell type and gene-specific thresholds. In hepatocellular carcinoma, the altered methylation patterns accompanying malignant transformation override the age-dependent increase in gene methylation. PMID- 15897562 TI - Genetic abnormalities and patterns of antigenic expression in multiple myeloma. AB - Myelomatous plasma cells show a high heterogeneity both in their immunophenotypic characteristics as well as in their cytogenetic features. Thus far, no extensive studies have been carried out to explore whether such antigenic diversity is associated with specific genetic characteristics. We have investigated the relationship between the immunophenotypic profile at plasma cell and both their DNA ploidy status (evaluated by flow cytometry) and specific genetic features (ascertained by fluorescence in situ hybridization) in a large series of 915 patients with newly diagnosed multiple myeloma. The non-hyperdiploid multiple myeloma group (n = 454, 52%) was associated with a significantly higher frequency of positivity for CD28 and CD20 as well as a higher incidence of CD56(-) and CD117(-) cases (P < 0.001). Remarkably, 13q deletion and immunoglobulin heavy chain (IGH) gene rearrangements, which were significantly more common in non hyperdiploid multiple myeloma, showed a strong association with CD117(-) cases. IGH translocation to 11q13 was associated with reactivity for CD20 (P < 0.001), down-regulation of CD56 (P < 0.001), and lack of expression of CD117 (P = 0.001). By contrast, IGH translocations to other chromosome partners were almost exclusively found among CD20(-) and CD117(-) cases (P < 0.001). These results suggest that genetic categories in multiple myeloma exhibit particular immunophenotypic profiles which in turn are strongly associated with the DNA ploidy status. PMID- 15897563 TI - Analysis of KIT mutations in sporadic and familial gastrointestinal stromal tumors: therapeutic implications through protein modeling. AB - PURPOSE: Gastrointestinal stromal tumors (GIST) are characterized by expressing a gain-of-function mutation in KIT, and to a lesser extent, PDGFR. Imatinib mesylate, a tyrosine kinase inhibitor, has activity against GISTs that contain oncogenic mutations of KIT. In this study, KIT and PDGFRalpha mutation status was analyzed and protein modeling approaches were used to assess the potential effect of KIT mutations in response to imatinib therapy. EXPERIMENTAL DESIGN: Genomic DNA was isolated from GIST tumors. Exons 9, 11, 13, and 17 of c-KIT and exons 12, 14, and 18 of PDGFRalpha were evaluated for oncogenic mutations. Protein modeling was used to assess mutations within the juxtamembrane region and the kinase domain of KIT. RESULTS: Mutations in KIT exons 9, 11, and 13 were identified in GISTs with the majority of changes involving the juxtamembrane region of KIT. Molecular modeling indicates that mutations in this region result in disruption of the KIT autoinhibited conformation, and lead to gain-of-function activation of the kinase. Furthermore, a novel germ-line mutation in KIT was identified that is associated with an autosomal dominant predisposition to the development of GIST. CONCLUSIONS: We have used protein modeling and structural analyses to elucidate why patients with GIST tumors containing exon 11 mutations are the most responsive to imatinib mesylate treatment. Importantly, mutations detected in this exon and others displayed constitutive activation of KIT. Furthermore, we have found tumors that are both KIT and PDGFRalpha mutation negative, suggesting that additional, yet unidentified, abnormalities may contribute to GIST tumorigenesis. PMID- 15897564 TI - Circulating tumor cells in breast cancer: correlation to bone marrow micrometastases, heterogeneous response to systemic therapy and low proliferative activity. AB - PURPOSE: The incidence and biological characteristics of circulating tumor cells in the blood of patients with breast cancer were examined and subgroups were evaluated in the context of systemic treatment and the presence of disseminated tumor cells in bone marrow. EXPERIMENTAL DESIGN: Circulating tumor cells were isolated from the peripheral blood of patients with breast cancer using a gradient system designed for the enrichment of circulating tumor cells (OncoQuick). Circulating tumor cells were identified with the anti-cytokeratin antibody, A45-B/B3. In subsets of patients, expression of the proliferation associated Ki-67 antigen in circulating tumor cells and the concomitant presence of micrometastases in bone marrow were examined. RESULTS: In patients with primary breast cancer (stage M(0)), circulating tumor cells were detected in 5 of 60 patients (8.3%) after surgery and before initiation of adjuvant chemotherapy; a positive correlation to the presence of disseminated tumor cells in bone marrow was observed (P = 0.030, n = 53). During the course of adjuvant chemotherapy, repeated analysis of 20 M(0) patients revealed the occurrence of circulating tumor cells in 7 of 16 patients that were initially negative. Patients with metastatic disease (stage M(1)) showed circulating tumor cells in 25 of 63 cases (39.7%, P < 0.0001 as compared with M(0) patients), and a positive finding was correlated with elevated concentrations of the serum tumor marker CA15.3 (P = 0.0093). Performing repeated analysis in a subgroup of 25 M(1) patients, circulating tumor cells were found more frequently in patients with progressive disease than in patients with stable disease or remission (87.5% versus 43.8% of patients with circulating tumor cells, respectively; P = 0.047). Independent of the disease-stage, none of the 47 patients examined for the proliferative status of their circulating tumor cells showed coexpression of Ki-67. CONCLUSIONS: Circulating tumor cells seem to be nonproliferating cells that persist during chemotherapy. Circulating tumor cell detection is linked to disease progression and elevated tumor marker concentrations in patients with metastatic breast cancer. PMID- 15897565 TI - Patterns of gene expression that characterize long-term survival in advanced stage serous ovarian cancers. AB - PURPOSE: A better understanding of the underlying biology of invasive serous ovarian cancer is critical for the development of early detection strategies and new therapeutics. The objective of this study was to define gene expression patterns associated with favorable survival. EXPERIMENTAL DESIGN: RNA from 65 serous ovarian cancers was analyzed using Affymetrix U133A microarrays. This included 54 stage III/IV cases (30 short-term survivors who lived <3 years and 24 long-term survivors who lived >7 years) and 11 stage I/II cases. Genes were screened on the basis of their level of and variability in expression, leaving 7,821 for use in developing a predictive model for survival. A composite predictive model was developed that combines Bayesian classification tree and multivariate discriminant models. Leave-one-out cross-validation was used to select and evaluate models. RESULTS: Patterns of genes were identified that distinguish short-term and long-term ovarian cancer survivors. The expression model developed for advanced stage disease classified all 11 early-stage ovarian cancers as long-term survivors. The MAL gene, which has been shown to confer resistance to cancer therapy, was most highly overexpressed in short-term survivors (3-fold compared with long-term survivors, and 29-fold compared with early-stage cases). These results suggest that gene expression patterns underlie differences in outcome, and an examination of the genes that provide this discrimination reveals that many are implicated in processes that define the malignant phenotype. CONCLUSIONS: Differences in survival of advanced ovarian cancers are reflected by distinct patterns of gene expression. This biological distinction is further emphasized by the finding that early-stage cancers share expression patterns with the advanced stage long-term survivors, suggesting a shared favorable biology. PMID- 15897566 TI - An innovative microarray strategy identities informative molecular markers for the detection of micrometastatic breast cancer. AB - There is increasing evidence that molecular detection of micrometastatic breast cancer in the axillary lymph nodes (ALN) of breast cancer patients can improve staging. Molecular analyses of samples obtained from the Minimally Invasive Molecular Staging of Breast Cancer Trial (n = 489 patients) indicate that whereas the majority of molecular markers are informative for the detection of metastatic breast cancer (significant disease burden), only a few are sensitive for the detection of micrometastatic disease (limited disease burden). Frequency distribution and linear regression analyses reveal that relative levels of gene expression are highly correlated with apparent sensitivity for the detection of micrometastic breast cancer (P < 0.05). These data provides statistical validation of the concept that the most informative markers for detection of micrometastatic disease are those that are most highly expressed in metastatic disease. To test this hypothesis, we developed an innovative microarray strategy. RNA from a metastatic breast cancer ALN was diluted into RNA from a normal lymph node and analyzed using Affymetrix microarrays. Expression analysis indicated that only two genes [mammaglobin (mam) and trefoil factor 1 (TFF1)] were significantly overexpressed at a dilution of 1:50. Real-time reverse transcription-PCR analysis of pathology-negative ALN (n = 72) confirm that of all the markers tested, mam and TFF1 have the highest apparent sensitivity for detection of micrometastatic breast cancer. We conclude that a dilutional microarray approach is a simple and reliable method for the identification of informative molecular markers for the detection of micrometastatic cancer. PMID- 15897567 TI - Tumor dose response to the vascular disrupting agent, 5,6-dimethylxanthenone-4 acetic acid, using in vivo magnetic resonance spectroscopy. AB - PURPOSE: To use (31)P and (1)H magnetic resonance spectroscopy (MRS) to assess changes in tumor metabolic profile in vivo in response to 5,6-dimethylxanthenone 4-acetic acid (DMXAA) with a view to identifying biomarkers associated with tumor dose response. EXPERIMENTAL DESIGN: In vivo (31)P and (1)H MRS measurements of (a) tumor bioenergetics [beta-nucleoside triphosphate/inorganic phosphate (beta NTP/Pi)], (b) the membrane-associated phosphodiesters and phosphomonoesters (PDE/PME), (c) choline (mmol/L), and (d) lactate/water ratio were made on murine HT29 colon carcinoma xenografts pretreatment and 6 or 24 hours posttreatment with increasing doses of DMXAA. Following in vivo MRS, the tumors were excised and used for high-resolution (31)P and (1)H MRS of extracts to provide validation of the in vivo MRS data, histologic analysis of necrosis, and high-performance liquid chromatography. RESULTS: Both beta-NTP/Pi and PDE/PME decreased in a dose dependent manner 6 hours posttreatment with DMXAA, with significant decreases in beta-NTP/Pi with 15 mg/kg (P < 0.001) and 21 mg/kg (P < 0.01). A significant decrease in total choline in vivo was found 24 hours posttreatment with 21 mg/kg DMXAA (P < 0.05); this was associated with a significant reduction in the concentration of the membrane degradation products glycerophosphoethanolamine and glycerophosphocholine measured in tissue extracts (P < 0.05). CONCLUSIONS: The reduction in tumor energetics and membrane turnover is consistent with the vascular-disrupting activity of DMXAA. (31)P MRS revealed tumor response to DMXAA at doses below the maximum tolerated dose for mice. Both (31)P and (1)H MRS provide biomarkers of tumor response to DMXAA that could be used in clinical trials. PMID- 15897568 TI - Carbonic anhydrase IX expression predicts outcome of interleukin 2 therapy for renal cancer. AB - PURPOSE: Renal cancer response to interleukin 2 (IL-2) therapy and patient survival has been correlated with tumor histology and carbonic anhydrase IX (CAIX) expression. In an effort to confirm and expand these observations, we examined CAIX expression in pathology specimens from renal cancer patients who had previously received IL-2 therapy. EXPERIMENTAL DESIGN: Paraffin-embedded tissue sections of renal cancer were immunostained with the MN-75 monoclonal antibody to CAIX and expression levels were correlated with histologic findings and clinical outcome. RESULTS: Tissue specimens were obtained from 66 patients; 27 of whom (41%) had responded to IL-2-based therapy. Fifty-eight specimens were assessed as clear cell, with 56, 33, and 4 having alveolar, granular, and papillary features, respectively. Twenty-four (36%), 31 (47%), and 11 (17%) were classified into good, intermediate, and poor prognosis groups according to the Upton pathology model. Forty-one specimens (62%) had high CAIX expression. Twenty one of 27 (78%) responding patients had high CAIX expressing tumors compared with 20 of 39 (51%) nonresponders (odds ratio, 3.3; P = 0.04). Median survival was prolonged (P = 0.04) and survival >5 years was only seen in high CAIX expressers. In patients with intermediate pathologic prognosis, all nine responders had high CAIX expression versus 11 of 22 nonresponders. A resultant group with good pathologic prognosis alone or with intermediate pathologic prognosis and high CAIX contained 26 of 27 (96%) responders compared with 18 of 39 (46%) nonresponders (odds ratio, 30; P < 0.01) and exhibited longer median survival (P < 0.01). CONCLUSIONS: CAIX expression seems to be an important predictor of outcome in renal cell carcinoma patients receiving IL-2-based therapy and may enhance prognostic information obtained from pathology specimens. PMID- 15897569 TI - Evaluation of a panel of tumor markers for molecular detection of circulating cancer cells in women with suspected breast cancer. AB - PURPOSE: We examined the feasibility of using molecular characterization of circulating tumor cells as a method for early detection of breast cancer. RESEARCH DESIGN: Women without a prior history of cancer who had a breast abnormality detected on imaging followed by a breast biopsy were enrolled in this study. Density gradient centrifugation and immunomagnetic capture were used to enrich for epithelial cells from approximately 20 mL of blood. Real-time reverse transcription-PCR was used to quantitate the expression levels of the highly breast-specific genes, mammaglobin, gamma-aminobutyric acid type A receptor pi subunit (GABA A(pi)), B305D-C, and B726P in the epithelial cell-enriched samples. RESULTS: The assay was technically feasible in 154 of 199 accrued patients. From their clinical assessment, 100 patients had benign breast disease, 10 patients had ductal carcinoma in situ, and 44 patients had invasive breast cancer. We constructed a diagnostic test that classified patients with mammaglobin levels of at least 32.2 copies/pg beta-actin (units) in their circulating epithelial cells as positive for invasive breast cancer. This resulted in a sensitivity and specificity of 63.3% and 75.0%, respectively. A diagnostic test that classified patients as positive for invasive breast cancer when either mammaglobin levels were >46.3 units or B305D-C levels were >11.6 units increased the sensitivity and specificity to 70.5% and 81.0%, respectively. In the latter test, 12 of the 14 node-positive breast cancer patients were correctly identified. Including GABA A(pi) and B726P in the test did not increase its diagnostic potential. CONCLUSIONS: These results suggest that molecular characterization of circulating epithelial cells using mammaglobin and B305D-C offers potential for early detection of invasive breast cancer. PMID- 15897570 TI - The relationship of molecular markers of p53 function and angiogenesis to prognosis of stage I epithelial ovarian cancer. AB - PURPOSE: Multiple angiogenic factors may influence tumor progression and metastasis. Several are modified by the p53 gene. We sought to identify molecular markers for high-risk stage I epithelial ovarian cancers. EXPERIMENTAL DESIGN: Seventy-seven consecutive stage I epithelial ovarian cancers were evaluated for p53, CD31 microvessel density, thrombospondin-1, vascular endothelial growth factor (VEGF), p21 immunohistochemical staining, and p53 gene mutations. Molecular marker impact upon disease-specific survival, disease recurrence, and distant recurrence was evaluated with Cox regression. RESULTS: There were 12 deaths from disease. Twelve of the 77 tumors contained p53 mutations-10 missense and 3 null (one tumor had two mutations). Fesddration Internationale des Gynaecologistes et Obstetristes substage (IA/IB versus IC; P < 0.001) and VEGF staining (P = 0.02) were significant in bivariate models with relationship to disease-specific survival. Stage (P = 0.0004), grade (P = 0.008), histology (P = 0.0025), p53 dysfunction (positive stain and/or mutation; P = 0.048), and microvessel density (P = 0.04) were significant in bivariate models with relationship to time to recurrence. In multivariate analyses among stage IC patients, failure to receive chemotherapy and microvessel density were associated with disease-specific survival, time to recurrence, and time to distant recurrence with hazard ratios of 4.8 to 44.1. CONCLUSIONS: The p53-dependent molecular markers of angiogenesis are of limited utility in developing a clinical strategy for postoperative management of stage I ovarian carcinoma. Microvessel density impacts survival and metastasis for high-risk stage IC disease. Adjuvant chemotherapy is necessary, but not sufficient, for cure of high-risk stage I epithelial ovarian cancers. PMID- 15897571 TI - Functionality of androgen receptor-based gene expression imaging in hormone refractory prostate cancer. AB - PURPOSE: A highly augmented, prostate-specific two-step transcriptional amplification (TSTA) method was developed with the ultimate goal of delivering an effective and safe gene-based treatment to prostate cancer patients. Because very limited treatment options are available for recurrent hormone refractory prostate cancer (HRPC), it is imperative to assess whether the prostate-specific antigen (PSA) promoter-based TSTA gene therapy will be functional in HRPC. EXPERIMENTAL DESIGN: We tested the TSTA-driven adenovirus vector on three androgen-dependent and six HRPC models. Real-time gene expression was monitored by both optical imaging and the combined modality of positron emission tomography (PET) and computed tomography. RESULTS: The TSTA-driven firefly luciferase expressing adenoviral vector was active in all androgen receptor (AR)-expressing HRPC models, but inactive in AR- and PSA-negative lines. Interestingly, the TSTA mediated gene expression was induced by hydrocortisone in MDA PCa 2b, a cell line with mutated AR that possesses altered ligand specificity. In animal models, the TSTA-mediated optical signal was more robust in the HRPC than androgen-dependent tumors. In a parallel trend, a TSTA vector that expresses the herpes simplex virus thymidine kinase PET reporter gene also displayed more robust PET signal in the HRPC tumor. CONCLUSIONS: The activity of TSTA system is AR dependent and it recapitulates the functional status of endogenous AR. These data support the conclusion that AR function is activated in HRPC despite castrated levels of androgen. Together with the fact that majority of recurrent prostate cancers express AR and PSA, we foresee that the TSTA approach can be a promising gene therapy strategy for the advanced stages of prostate cancer. PMID- 15897572 TI - Mutation in the tyrosine kinase domain of epidermal growth factor receptor is a predictive and prognostic factor for gefitinib treatment in patients with non small cell lung cancer. AB - PURPOSE: Mutations in epidermal growth factor receptor (EGFR) can be used to predict the tumor response of patients receiving gefitinib for non-small cell lung cancer (NSCLC). We investigated the association between mutations in EGFR tyrosine kinase domain and tumor response and survival in gefitinib-treated NSCLC patients. EXPERIMENTAL DESIGN: EGFR mutations in exons 18 to 21 were analyzed by DNA sequencing of paraffin-embedded tumor tissues from gefitinib-treated NSCLC patients. The results were correlated with clinical variables. RESULTS: EGFR mutations were found in 61.1% (33 of 54) of cases; response rate and disease control rate were 56.8% and 68.5%, respectively. There was no significant difference in mutation rates between adenocarcinoma (29 of 43) and nonadenocarcinoma (4 of 11; P = 0.085). However, all four nonadenocarcinomas with EGFR mutations had no response to gefitinib. Presence of EGFR mutations was the only independent predictor for disease control (P = 0.003) and tumor response (P = 0.017) in multivariate analysis; positive predictive values were 87.9% and 70.8% and negative predictive values were 61.9% and 69.2%, respectively. In comparison with patients whose tumor was negative for EGFR mutations, patients with EGFR mutations had better progression-free survival (median, 7.6 versus 1.7 months; P = 0.011) and overall survival (median, 14.7 versus 4.7 months; P = 0.046). CONCLUSIONS: Mutations in EGFR tyrosine kinase correlate with treatment response and survival in gefitinib-treated NSCLC patients and can be used as a predictive and prognostic factor. Thus, analysis of EGFR tyrosine kinase mutations in lung adenocarcinoma is of clinical significance, as it can permit the customization of treatment with EGFR tyrosine kinase inhibitors. PMID- 15897573 TI - Cytochrome p450 profile of colorectal cancer: identification of markers of prognosis. AB - PURPOSE: The cytochromes P450 (P450) are a multigene family of enzymes with a central role in the oxidative metabolism of a wide range of xenobiotics, including anticancer drugs, carcinogens, and endogenous compounds. The purpose of this study was to define the P450 profile of colorectal cancer and establish the prognostic significance of expression of individual P450s in colorectal cancer. EXPERIMENTAL DESIGN: Immunohistochemistry for a panel of 23 P450s was done on a colorectal cancer tissue microarray consisting of 264 primary colorectal cancers, 91 lymph node metastasis, and 10 normal colorectal samples. The intensity of immunoreactivity in each sample was established by light microscopy. RESULTS: The most frequently expressed form of P450 in normal colon was CYP3A4. In primary colorectal cancer, several P450s (CYP1B1, CYP2S1, CYP2U1, CYP3A5, and CYP51) were present at a significantly higher level of intensity compared with normal colon. P450 expression was also detected in lymph node metastasis and the presence of several P450s (CYP1B1, CYP2A/2B, CYP2F1, CYP4V2, and CYP39) in the lymph node metastasis strongly correlated with their presence in corresponding primary tumors. The presence of strong CYP51 (log-rank = 12.11, P = 0.0005) or strong CYP2S1 (log-rank = 6.72, P = 0.0095) immunoreactivity were associated with poor prognosis. CYP51 was also an independent marker of prognosis (P = 0.009). CONCLUSIONS: The expression of individual P450s has been established in colorectal cancer. Several P450s show increased expression in colorectal cancer. High expression of CYP51 or CYP2S1 were associated with poor prognosis and CYP51 is an independent marker of prognosis. PMID- 15897574 TI - Markers of adenocarcinoma characteristic of the site of origin: development of a diagnostic algorithm. AB - PURPOSE: Patients with metastatic adenocarcinoma of unknown origin are a common clinical problem. Knowledge of the primary site is important for their management, but histologically, such tumors appear similar. Better diagnostic markers are needed to enable the assignment of metastases to likely sites of origin on pathologic samples. EXPERIMENTAL DESIGN: Expression profiling of 27 candidate markers was done using tissue microarrays and immunohistochemistry. In the first (training) round, we studied 352 primary adenocarcinomas, from seven main sites (breast, colon, lung, ovary, pancreas, prostate and stomach) and their differential diagnoses. Data were analyzed in Microsoft Access and the Rosetta system, and used to develop a classification scheme. In the second (validation) round, we studied 100 primary adenocarcinomas and 30 paired metastases. RESULTS: In the first round, we generated expression profiles for all 27 candidate markers in each of the seven main primary sites. Data analysis led to a simplified diagnostic panel and decision tree containing 10 markers only: CA125, CDX2, cytokeratins 7 and 20, estrogen receptor, gross cystic disease fluid protein 15, lysozyme, mesothelin, prostate-specific antigen, and thyroid transcription factor 1. Applying the panel and tree to the original data provided correct classification in 88%. The 10 markers and diagnostic algorithm were then tested in a second, independent, set of primary and metastatic tumors and again 88% were correctly classified. CONCLUSIONS: This classification scheme should enable better prediction on biopsy material of the primary site in patients with metastatic adenocarcinoma of unknown origin, leading to improved management and therapy. PMID- 15897575 TI - Detection of micrometastases in pelvic lymph nodes in patients undergoing radical cystectomy for locally invasive bladder cancer by real-time reverse transcriptase PCR for cytokeratin 19 and uroplakin II. AB - PURPOSE: The objective of this study was to clarify the significance of micrometastases in pelvic lymph nodes in patients who underwent radical cystectomy for bladder cancer. EXPERIMENTAL DESIGN: We included 40 patients with locally invasive bladder cancer who underwent radical cystectomy and pelvic lymphadenectomy. Expression of cytokeratin 19 (CK19), uroplakin II (UP II), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in 760 lymph nodes were assessed by a fully quantitative real-time reverse transcription-PCR (RT-PCR) assay. The quantification value of CK19 or UP II mRNA was described as each value relative to GAPDH mRNA. In this study, we regarded specimen in which either CK19 or UP II mRNA was positive as "presence of micrometastasis." RESULTS: Routine pathologic examinations detected tumor cells in 29 lymph nodes from six patients. Real-time RT-PCR identified positive expression of CK19 and UP II mRNAs in 49 lymph nodes from 10 patients and 98 lymph nodes from 16 patients, respectively. Of 633 lymph nodes from 34 patients with no pathologic evidence of nodal involvement, 13 nodes from five patients and 58 nodes from 10 patients were diagnosed as positive for CK19 and UP II mRNAs expression, respectively, by real-time RT-PCR. Presence of micrometastases was significantly associated with other conventional prognostic variables, including pathologic stage and microvascular invasion. Disease recurrence was occurred in eight patients, among whom four patients were negative for lymph node metastasis by routine pathologic examination and diagnosed as having micrometastasis by real-time RT-PCR assay. Furthermore, cause-specific survival rate in patients without micrometastasis was significantly higher than that in those with micrometastasis, irrespective of the presence of pathologic positive nodes. CONCLUSIONS: Approximately 30% of locally invasive bladder cancer shed cancer cells to pelvic lymph nodes, and disease recurrence after radical cystectomy could be explained, at least in part, by micrometastases in pelvic lymph nodes. PMID- 15897576 TI - Prognostic role of thymidylate synthase polymorphisms in gastric cancer patients treated with surgery and adjuvant chemotherapy. AB - PURPOSE: To investigate the prognostic role of thymidylate synthase (TS) polymorphisms in gastric cancer patients treated with radical surgery and fluorouracil-based adjuvant chemotherapy. EXPERIMENTAL DESIGN: Ninety gastric cancer cases were identified among 187 patients previously enrolled in prospective case-control studies for disease susceptibility. Patients were genotyped for a G/C nucleotide change within a triple 28 bp variable number of tandem repeat sequence in the TS 5'-untranslated region (5'-UTR) and a 6 bp deletion in the TS 3'-untranslated region (3'-UTR). According to available functional data, patients with 5'-UTR 2R/2R, 2R/3C, 3C/3C genotypes were classified as low TS producers (5'-UTRlow) and patients with 5'-UTR 3G/3G, 3G/3C, 2R/3G genotypes as high TS producers (5'UTRhigh). Patients with 3'-UTR del6/del6 and del6/ins6 genotypes were classified as low TS producers (3'-UTRlow) and patients with 3'-UTR ins6/ins6 genotype as high TS producers (3'-UTRhigh). The prognostic analysis was based on 5'-UTR/3'-UTR combined genotypes. RESULTS: Ten patients (11%) were 5'-UTRhigh/3'-UTRhigh, 36 patients were 5'-UTRhigh/3'-UTRlow, 19 patients were 5'-UTRlow/3'-UTRhigh, and 25 patients were 5'-UTRlow/3'-UTRlow. 5'-UTRlow/3'-UTRlow patients showed the best outcome and the threshold of statistical significance was achieved in the comparison of disease-free survival and overall survival with 5'-UTRhigh/3'-UTRlow patients and 5'-UTRhigh/3'-UTRhigh patients. The presence of at least one high TS expression genotype showed independent adverse prognostic role in multivariate analysis. CONCLUSIONS: The prognostic role of TS polymorphisms in gastric cancer deserves further investigation because the adverse effect of high TS expression genotypes may be a relevant information to improve adjuvant chemotherapeutic strategies. PMID- 15897577 TI - Cyclooxygenase-2 overexpression is associated with a poor outcome in resected ampullary cancer patients. AB - PURPOSE: To identify potential prognostic molecular factors in ampullary adenocarcinoma that could be of significant importance. To this end, we examined the possible prognostic significance of cyclooxygenase-2 (Cox-2) and Survivin expression and the apoptotic index in a cohort of uniformly treated patients with ampullary cancer treated with radical surgical excision. EXPERIMENTAL DESIGN: The entry criteria were that the patients have a pathologic diagnosis of ampullary cancer which had been resected. Expression analysis for Cox-2 and Survivin was done by immunohistochemical staining. Apoptotic cells were identified by the terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method. RESULTS: Thirty-nine tumor specimens from resected ampullary adenocarcinoma patients were included. By univariate analysis, overall survival was affected by Cox-2 expression and TUNEL staining (respectively, P = 0.0003 and 0.03). Survivin expression did not influence the overall survival in our patient population (P = 0.123). Patients' clinicopathologic features (gender, age, and T and N factors) did not influence outcome. In multivariate Cox regression analysis, Cox-2 expression (relative risk, 4.330; P = 0.005) was the only variable that significantly affected overall survival. CONCLUSIONS: The results of the present article provide, for the first time, evidence that Cox-2 expression, but not Survivin expression, may represent a significant prognostic factor after surgical resection in patients affected by cancer of the ampulla of Vater. Further studies are required to determine whether Cox-2 inhibitors may be useful for the therapy or prevention of ampullary carcinoma. PMID- 15897578 TI - Inhibition of heme oxygenase-1 increases responsiveness of pancreatic cancer cells to anticancer treatment. AB - Heme oxygenase-1 (HO-1) is believed to represent a key enzyme for the protection of cells against "stress." Its overexpression in different types of human cancers supports the notion that HO-1 provides a growth advantage and contributes to cellular resistance against chemotherapy and radiotherapy. Given the poor survival rates of patients with pancreatic cancer due to its aggressive growth behavior and its exceptional resistance to all known forms of anticancer treatment, we have investigated the expression of HO-1 in human pancreatic cancer cells growth behavior and prognosis. Expression of HO-1 was analyzed in human pancreatic cancer samples in comparison with normal pancreas by quantitative PCR, Western blot, and confocal microscopy. The influence of radiotherapy and chemotherapy on HO-1 expression in pancreatic cancer cell lines was evaluated. Furthermore, HO-1 expression was specifically suppressed by small interfering RNA transfection and subsequently the alterations of growth behavior and resistance to anticancer treatment were tested. Human pancreatic cancer showed a 6-fold and 3.5-fold HO-1 up-regulation in comparison to normal pancreas based on mRNA and protein level, respectively (P < 0.05). Cancer tissues revealed marked HO-1 immunoreactivity in tumor cells and in tumor associated immunocytes. Treatment of the pancreatic cancer cell lines with gemcitabine or radiation strongly induced HO-1 expression. Targeted knockdown of HO-1 expression led to pronounced growth inhibition of the pancreatic cancer cells and made tumor cells significantly more sensitive to radiotherapy and chemotherapy. Therefore, specific inhibition of HO 1 expression may be a new option in pancreatic cancer therapy and may be used as sensitizer to chemotherapy and radiotherapy. PMID- 15897579 TI - Proton beam therapy for hepatocellular carcinoma: a retrospective review of 162 patients. AB - PURPOSE: We present results of patients with hepatocellular carcinoma (HCC) treated with proton beam therapy. EXPERIMENTAL DESIGN: We reviewed 162 patients having 192 HCCs treated from November 1985 to July 1998 by proton beam therapy with or without transarterial embolization and percutaneous ethanol injection. The patients in the present series were considered unsuitable for surgery for various reasons, including hepatic dysfunction, multiple tumors, recurrence after surgical resection, and concomitant illnesses. The median total dose of proton irradiation was 72 Gy in 16 fractions over 29 days. RESULTS: The overall survival rate for all of the 162 patients was 23.5% at 5 years. The local control rate at 5 years was 86.9% for all 192 tumors among the 162 patients. The degree of impairment of hepatic functions attributable to coexisting liver cirrhosis and the number of tumors in the liver significantly affected patient survival. For 50 patients having least impaired hepatic functions and a solitary tumor, the survival rate at 5 years was 53.5%. The patients had very few acute reactions to treatments and a few late sequelae during and after the treatments. CONCLUSIONS: Proton beam therapy for patients with HCC is effective, safe, well tolerable, and repeatable. It is the useful treatment mode for either cure or palliation for patients with HCC irrespective of tumor size, tumor location in the liver, insufficient feeding of the tumor with arteries, presence of vascular invasion, impaired hepatic functions, and coexisting intercurrent diseases. PMID- 15897580 TI - Phase I and pharmacokinetic study of the dolastatin 10 analogue TZT-1027, given on days 1 and 8 of a 3-week cycle in patients with advanced solid tumors. AB - PURPOSE: TZT-1027 [N(2)-(N,N-dimethyl-l-valyl)-N-[(1S,2R)-2-methoxy-4-[(2S)-2 [(1R,2R)-1-methoxy-2-methyl-3-oxo-3-[(2-phenylethyl)]amino]propyl]-1 pyrrolidinyl]-1-[(S)-1-methylpropyl]-4-oxobutyl]-N-methyl-l-valinamide] is a cytotoxic dolastatin 10 derivative inhibiting microtubule assembly through the binding to tubulins. The objectives of this phase I study was to assess the dose limiting toxicities (DLT), to determine the maximum tolerated dose, and to study the pharmacokinetics of TZT-1027 when given i.v. over 60 minutes on days 1 and 8 every 3 weeks to patients with advanced solid tumors. EXPERIMENTAL DESIGN: Patients were treated with escalating doses of TZT-1027 at doses ranging from 1.35 to 2.7 mg/m(2). For pharmacokinetic analysis, plasma sampling was done during the first and second course and assayed using a validated high-performance liquid chromatographic assay with mass spectrometric detection. RESULTS: Seventeen patients received a total of >70 courses. The stopping dose was reached at 2.7 mg/m(2), with neutropenia and infusion arm pain as DLT. Neutropenia was not complicated by fever. Over all dose levels, eight patients experienced pain in the infusion arm 1 to 2 days after administration of the drug, which seemed ameliorated by adding additional flushing after drug administration. Other side effects included nausea, vomiting, diarrhea, and fatigue. One partial response lasting >54 weeks was observed in an extensively pretreated patient with metastatic liposarcoma. The pharmacokinetics of TZT-1027 suggested linearity over the dose ranges. No correlation between body surface area and absolute CL of TZT 1027 was established, vindicating that a flat dosing regimen might be used in the future. A correlation was observed between the percentage decrease in neutrophil count and the AUC of TZT-1027. CONCLUSIONS: In this study, the DLT of TZT-1027 was neutropenia and infusion arm pain. The recommended dose for phase II studies of TZT-1027 is 2.4 mg/m(2) given i.v. over 60 minutes, on days 1 and 8 every 21 days. Phase II studies have recently started. PMID- 15897581 TI - Humoral immune response to mesothelin in mesothelioma and ovarian cancer patients. AB - PURPOSE: Mesothelin is a glycosyl-phosphatidylinositol-anchored glycoprotein present on the cell surface. Mesothelin is a differentiation antigen that is highly expressed on mesothelioma, ovarian cancer, and pancreatic cancer. The existence of a spontaneous humoral immune response to mesothelin in humans has not been fully studied. Here we addressed the issue of whether mesothelin elicits a humoral immune response in patients with mesothelioma and ovarian cancer. EXPERIMENTAL DESIGN: Using an ELISA, we analyzed immunoglobulin G antibodies specific for mesothelin in sera from patients with mesothelioma and epithelial ovarian cancer. Tumor specimens were examined by immunohistochemistry for mesothelin protein expression. RESULTS: Elevated levels of mesothelin-specific antibodies were detected in the sera of 39.1% of patients with mesothelioma (27 of 69 patients) and 41.7% with epithelial ovarian cancer (10 of 24 patients) when compared with a normal control population (44 blood donors; P < 0.01 for both mesothelioma and ovarian cancer). We also found that 53% to 56% of patients with mesothelin immunostaining-positive mesothelioma and ovarian cancer had antibodies specific for mesothelin, whereas only 0% to 8% of patients with negative mesothelin immunostaining had detectable mesothelin-specific antibodies (chi(2) test: P < 0.01 for mesothelioma and P = 0.025 for ovarian cancer). CONCLUSIONS: Our findings indicate that mesothelin is a new tumor antigen in patients with mesothelioma and ovarian cancer and the immunogenicity of mesothelin is associated with its high expression on the tumor cells. Mesothelin represents an excellent target for immune-based therapies. PMID- 15897582 TI - Recombinant Sendai virus vector induces complete remission of established brain tumors through efficient interleukin-2 gene transfer in vaccinated rats. AB - PURPOSE: Sendai virus (SeV), a murine parainfluenza virus type I, replicates independent of cellular genome and directs high-level gene expressions when used as a viral vector. We constructed a nontransmissible recombinant SeV vector by deleting the matrix (M) and fusion (F) genes from its genome (SeV/DeltaMDeltaF) to enhance its safety. We also estimated the therapeutic efficacy of the novel vector system against a rat glioblastoma model. EXPERIMENTAL DESIGN: We administered the recombinant SeV vector carrying the lacZ gene or the human interleukin-2 (hIL-2) gene into established 9L brain tumors in vivo simultaneous with peripheral vaccination using irradiated 9L cells. Sequential monitoring with magnetic resonance imaging was used to evaluate the therapeutic efficacy. RESULTS: We found extensive transduction of the lacZ gene into the brain tumors and confirmed sufficient amounts of interleukin 2 (IL-2) production by hIL2 SeV/DeltaMDeltaF both in vitro and in vivo. The magnetic resonance imaging study showed that the intracerebral injection of hIL2-SeV/DeltaMDeltaF brought about significant reduction of the tumor growth, including complete elimination of the established brain tumors. The (51)Cr release assay showed that significant amounts of 9L-specific cytotoxic T cells were induced by the peripheral vaccination. Immunohistochemical analysis revealed that CD4(+) T cells and CD8(+) T cells were abundantly infiltrated in the target tumors. CONCLUSION: The present results show that the recombinant nontransmissible SeV vector provides efficient in vivo gene transfer that induces significant regression of the established brain tumors and suggest that it will be a safe and useful viral vector for the clinical practice of glioma gene therapy. PMID- 15897583 TI - Dietary flaxseed alters tumor biological markers in postmenopausal breast cancer. AB - PURPOSE: Flaxseed, the richest source of mammalian lignan precursors, has previously been shown to reduce the growth of tumors in rats. This study examined, in a randomized double-blind placebo-controlled clinical trial, the effects of dietary flaxseed on tumor biological markers and urinary lignan excretion in postmenopausal patients with newly diagnosed breast cancer. EXPERIMENTAL DESIGN: Patients were randomized to daily intake of either a 25 g flaxseed-containing muffin (n = 19) or a control (placebo) muffin (n = 13). At the time of diagnosis and again at definitive surgery, tumor tissue was analyzed for the rate of tumor cell proliferation (Ki-67 labeling index, primary end point), apoptosis, c-erbB2 expression, and estrogen and progesterone receptor levels. Twenty-four-hour urine samples were analyzed for lignans, and 3-day diet records were evaluated for macronutrient and caloric intake. Mean treatment times were 39 and 32 days in the placebo and flaxseed groups, respectively. RESULTS: Reductions in Ki-67 labeling index (34.2%; P = 0.001) and in c-erbB2 expression (71.0%; P = 0.003) and an increase in apoptosis (30.7%; P = 0.007) were observed in the flaxseed, but not in the placebo group. No significant differences in caloric and macronutrient intake were seen between groups and between pre- and posttreatment periods. A significant increase in mean urinary lignan excretion was observed in the flaxseed group (1,300%; P < 0.01) compared with placebo controls. The total intake of flaxseed was correlated with changes in c-erbB2 score (r = -0.373; P = 0.036) and apoptotic index (r = 0.495; P < 0.004). CONCLUSION: Dietary flaxseed has the potential to reduce tumor growth in patients with breast cancer. PMID- 15897584 TI - A phase I clinical trial of the sequential combination of irinotecan followed by flavopiridol. AB - PURPOSE: Flavopiridol potently enhances the effect of irinotecan with cures in colorectal cancer xenografts, and is associated with modulation of several molecular targets, including p21, Differentiation-related gene 1 (Drg1), and p53. We initiated a phase I trial of the sequential combination of irinotecan followed by flavopiridol to determine the maximal tolerated dose of this combination therapy. PATIENTS AND METHODS: Forty-five patients with advanced solid tumors were enrolled. Irinotecan was administered first (100 or 125 mg/m(2)) followed 7 hours later by escalating flavopiridol (10-70 mg/m(2)) given weekly over 1 hour for 4 of 6 weeks. At the maximal tolerated dose, the pharmacokinetic analysis was expanded and pre- and posttreatment tumor biopsies were done. RESULTS: At irinotecan 100 mg/m(2), dose-limiting diarrhea and myelosuppression were observed with flavopiridol 70 mg/m(2). At irinotecan 125 mg/m(2), we observed dose limiting hyperbilirubinemia, fatigue, and myelosuppression at flavopiridol 60 mg/m(2). Peak flavopiridol concentrations of >/=2 mumol/L were achieved above flavopiridol 50 mg/m(2). No significant pharmacokinetic interactions with irinotecan were noted. Baseline serum bilirubin significantly predicted cycle 1 dose-limiting toxicity and neutropenia. We observed partial responses in three patients and prolonged stable disease (i.e., >6 months) in 36% of patients including adrenocortical cancer and hepatocellular cancer. Patients with wild type p53 and either no change or low posttreatment biopsy p21 and a decrease in Drg1 expression showed stable or responsive disease to the combination therapy. CONCLUSIONS: The recommended phase II dose with irinotecan 100 mg/m(2) is flavopiridol 60 mg/m(2) and with irinotecan 125 mg/m(2) is flavopiridol 50 mg/m(2). Toxicity can be predicted by baseline bilirubin. Clinical activity is encouraging and may correlate to changes in p21 and Drg1 levels in patients with wild type p53 tumors following therapy. PMID- 15897585 TI - Phase 1 clinical and pharmacokinetics evaluation of oral CI-1033 in patients with refractory cancer. AB - PURPOSE: To determine the tolerability and pharmacokinetics of oral CI-1033, a pan-erbB tyrosine kinase inhibitor, administered over 14 consecutive days of a 21 day cycle. DESIGN: Phase 1, multicenter trial involving patients with solid tumors that are refractory to standard therapy. CI-1033 was administered initially at 300 mg/day to a minimum cohort of three patients. Dose escalation proceeded at LDLR-/-; n = 19) or wild-type C57BL/6 littermates (C57 BL/6-->LDLR-/-; n = 19) and maintained for 8 weeks on chow and then for 9 weeks on a Western-type diet. Plasma sPLA2 activity and plasma lipoprotein profiles were not significantly different between sPLA2-->LDLR /- and C57BL/6-->LDLR-/- mice. Aortic root atherosclerosis was increased by 57% in sPLA2-->LDLR-/- mice compared with C57BL/6-->LDLR-/- controls (P < 0.05). Foam cell formation in vitro and in vivo was increased significantly. Urinary, plasma, and aortic levels of the isoprostane 8,12-iso-iPF2alpha-VI and aortic levels of 12/15-LO reaction products were each significantly higher (P < 0.001) in sPLA2- >LDLR-/- compared with C57BL/6-->LDLR-/- mice, indicating significantly increased in vivo oxidative stress in sPLA2--> LDLR-/-. These data demonstrate that macrophage-specific overexpression of human sPLA2 increases atherogenesis by directly modulating foam cell formation and in vivo oxidative stress without any effect on systemic sPLA2 activity and lipoprotein metabolism. PMID- 15897608 TI - A targeted mass spectrometric analysis of phosphatidylinositol phosphate species. AB - The development of a new mass spectrometric lipid profiling methodology permits the identification of cellular phosphatidylinositol monophosphate/phosphatidylinositol bisphosphate/phosphatidylinositol trisphosphate (PIP/PIP2/PIP3) species that includes the fatty acyl composition. Using electrospray ionization mass spectrometry, we were able to resolve and identify 28 PIP and PIP2 compounds as well as 8 PIP3 compounds from RAW 264.7 or primary murine macrophage cell extracts. Analysis of PIP profiles after agonist stimulation of cells revealed the generation of differential PIP3 species and permitted us to propose a novel means for regulation and specificity in signaling through PIP3. This is the first reported identification of intact, cellular PIP3 by mass spectral analysis. The ability to analyze the fatty acyl chain composition of signaling lipids initiates new venues for investigation of the processes by which specific polyphosphoinositide species mediate. PMID- 15897609 TI - Transfer of cholesteryl esters and phospholipids as well as net deposition by microsomal triglyceride transfer protein. AB - Microsomal triglyceride transfer protein (MTP) activity is classically measured using radioactive lipids. We described a simple fluorescence assay to measure its triacylglycerol (TAG) transfer activity. Here, we describe fluorescence-based methods to measure the transfer of phospholipids (PLs) and cholesteryl esters (CEs) by MTP. Both transfer activities increased with time and MTP amounts and were inhibited to different extents by an MTP antagonist, BMS197636. We also describe a method to measure the net deposition of fluorescent lipids in acceptor vesicles. In this procedure, negatively charged donor vesicles are incubated with MTP and acceptor vesicles, and lipids transferred to acceptors are quantified after the removal of donor vesicles and MTP by the addition of DE52. Lipid deposition in acceptor vesicles was dependent on time and MTP. Using these methods, TAG transfer activity was the most robust activity present in purified MTP; CE and PL transfer activities were 60-71% and 5-13% of the TAG transfer activity, respectively. The method to determine lipid transfer is recommended for routine MTP activity measurements for its simplicity. These methods may help identify specific inhibitors for individual lipid transfer activities, in characterizing different domains involved in transfer, and in the isolation of mutants that bind but cannot transfer lipids. PMID- 15897610 TI - Administration of a PPARalpha agonist increases serum apolipoprotein A-V levels and the apolipoprotein A-V/apolipoprotein C-III ratio. AB - Apolipoprotein A-V (apoA-V) first gained attention as a regulator of triglycerides through transgenic mouse studies. Furthermore, peroxisome proliferator-activated receptor alpha (PPARalpha) agonists such as fenofibrate increase apoA-V mRNA expression. Our group recently developed the first assay to quantitate serum apoA-V levels. Therefore, we sought to determine whether administration of a PPARalpha agonist would increase circulating apoA-V. Cynomolgus monkeys were dosed for 14 days with 0.3 mg/kg/day LY570977 L-lysine, a potent and selective PPARalpha agonist. Blood samples were drawn throughout the treatment period and after a 2 week washout. Administration of the PPARalpha agonist caused a 50% decrease in triglycerides that reversed at washout. Serum apoA-V concentrations increased 2-fold, correlated inversely with triglycerides, and were reversible at washout. The apoA-V/apoC-III ratio increased >2-fold, with this increase also reversible at washout. These data demonstrate for the first time that a PPARalpha agonist increases circulating apoA-V protein levels and the apoA-V/apoC-III ratio. PMID- 15897611 TI - Fundamentals of modern peptide synthesis. AB - The purpose of this chapter is to delineate strategic considerations and provide practical procedures to enable non-experts to synthesize peptides with a reasonable chance of success. This chapter focuses on Fmoc chemistry, which is now the most commonly employed strategy for solid phase peptide synthesis (SPPS). Protocols for the synthesis of fully deprotected peptides are presented, together with a review of linkers and supports currently employed for SPPS. The principles and the different steps of SPPS (anchoring, deprotection, coupling reaction, and cleavage) are all discussed, along with their possible side reactions. PMID- 15897612 TI - Looking forward to open access. PMID- 15897613 TI - Elsevier and open access. PMID- 15897614 TI - Necessity is the mother of innovation. PMID- 15897615 TI - Brain architecture management system. AB - The nervous system can be viewed as a biological computer whose genetically determined macrocircuitry has two basic classes of parts: gray matter regions interconnected by fiber pathways. We describe here the basic features of an online knowledge management system for storing and inferring relationships between data about the structural organization of nervous system circuitry. It is called the Brain architecture management system (BAMS; http://brancusi.usc.edu/bkms) and it stores and analyzes data specifically concerned with nomenclature and its hierarchical taxonomy, with axonal connections between regions, and with the neuronal cell types that form regions and fiber pathways. PMID- 15897616 TI - A modeling environment with three-dimensional morphology, A-Cell-3D, and Ca2+ dynamics in a spine. AB - A-Cell-3D was developed to model and simulate a neuron with three-dimensional (3D) morphology utilizing graphic user interface (GUI)-based operations. A-Cell 3D generates and compartmentalizes 3D morphologies of a whole cell or a part of a cell based on a small number of parameters. A-Cell-3D has functions for embedding biochemical reactions and electrical equivalent circuits in the generated 3D morphology, automatically generating a simulation program for spatiotemporal numerical integration, and for visualizing the simulation results. A-Cell-3D is a free software and will be a powerful tool for both experimental and theoretical researchers in modeling and simulating neurons. The Ca2+ dynamics in a dendritic spine and its parent dendrite were modeled and simulated to demonstrate the capabilities of A-Cell-3D. The constructed reaction-diffusion model comprised Ca2+ entry at the spine head, Ca2+ buffering by endogenous buffers, Ca2+ extrusion, and Ca2+ diffusion with or without exogenous Ca2+ indicator dyes. A simulation program was generated by A-Cell-3D, and differential equations were numerically integrated by the fourth-order Runge-Kutta method. PMID- 15897617 TI - BrainMap: the social evolution of a human brain mapping database. AB - Human brain mapping is an experimental discipline that establishes structure function correspondences in the brain through the combined application of experimental psychology, human neuroscience, and noninvasive neuroimaging. A deep and diverse literature on the functional organization of the human brain is emerging, which has pushed neuroimaging squarely into the scientific mainstream. Because of this rapid growth, there is a great need to effectively collect and synthesize the body of literature in this field. The BrainMap database was created in response to this need as an electronic environment for modeling the human brain through quantitative meta-analysis of the brain mapping literature. BrainMap was originally conceived in 1987 and has received continuous funding from 1988 to 2004. During this time, BrainMap has consistently evolved to meet the challenges of an ever-changing field and continues to strive toward higher levels of applicability. In this article, we discuss BrainMap's structure and utility, and relate its progress and development as a neuroinformatics tool. PMID- 15897618 TI - Identification of a major gene responsible for type 1 diabetes in the Komeda diabetes-prone rat. AB - Type 1 diabetes mellitus is an autoimmune disease involving both environmental and genetic factors. Genetic analyses in humans and rodents have shown that the major histocompatibility complex (MHC) is a major genetic factor and that several other genes may be involved in the development of the disease. We performed genetic analysis of type 1 diabetes in a newly established animal model, the Komeda diabetes-prone (KDP) rat, and found that most of the genetic predisposition to diabetes is accounted for by two major susceptibility genes, MHC and Iddm/kdp1. In addition, we identified a nonsense mutation in the Casitas B-lineage lymphoma b (Cblb) gene by positional cloning of Iddm/kdp1. In this paper, I review our positional cloning analysis of Iddm/kdp1 and propose a two gene model of the development of type 1 diabetes in which two major susceptibility genes, Cblb and MHC, determine autoimmune reaction and tissue specificity to pancreatic beta-cells, respectively. PMID- 15897619 TI - A simple method of intestinal anastomosis (ileocolostomy) in rats. AB - A simple method of ileocolostomy was performed in rats. The colon near the cecum was ligated, including its arteries and veins. Main artery and vein of the cecum were ligated. Then, the cecum was cut out. A longitudinal incision was made on the anti-mesenteric side of the proximal end of the colon, approximately 7-8 mm long. A 21-G needle was inserted toward the incision 2 cm away from the proximal end of the anti-mesenteric side of the colon. A nylon suture was knotted once to the distal end of the ileum and was introduced into the tip of the needle which had previously been passed through the colon. Then, the needle was removed. The suture was pulled to introduce the distal end of the ileum into the colonic lumen. Then, the suture was knotted once on the colon again to fix the ileum to the colon. The incision in the proximal end of the colon was not closed. At the 2nd week after the operation, X-ray examinations demonstrated that the ileocolonic passages with no leakage at the anastomotic site were quite satisfactory. At the 4th week after the operation, there were no macroscopic or microscopic complications at the anastomotic site. The mucosal and serosal epithelia of the ileum and colon continued smoothly. This simple method may be very effective in preparing anastomosis in the gastrointestinal tract, especially in small laboratory animals for nutritional and surgical experiments. PMID- 15897620 TI - Identification procedure for Pasteurella pneumotropica in microbiologic monitoring of laboratory animals. AB - Discrepancies have been recognized in the identification of Pasteurella pneumotropica between testing laboratories. To determine the causes of the differences and to propose a reliable identification procedure for P. pneumotropica, a working group was organized and 69 isolates identified or suspected as P. pneumotropica were collected from 8 laboratories in Japan. These isolates were examined by colony morphology, Gram-staining, the slide agglutination test using two antisera (ATCC35149 and MaR), two commercially available biochemical test kits (ID test, API20NE) and two primer sets of PCR tests (Wang PCR, CIEA PCR). The 69 isolates and two reference strains were divided into 10 groups by test results. No single procedure for P. pneumotropica identification was found. Among tested isolates, large differences were not observed by colony morphology and Gram-straining except for colony colors that depended on their biotypes. Sixty-eight out of 69 isolates were positive by the slide agglutination test using two antisera except for one isolate that tested with one antiserum. The ID test identified 61 out of 69 isolates as P. pneumotropica and there was no large difference from the results of CIEA PCR. From these results, we recommend the combination of colony observation, Gram straining, the slide agglutination tests with two antisera and biochemical test using the ID test for practical and reliable identification of this organism. PMID- 15897621 TI - No regional differences of cytochrome P450 expression in the liver of Cynomolgus monkeys (Macaca fascicularis). AB - Non-human primates are frequently used in toxicological studies the result of which are extrapolated to humans, but background data on drug metabolism ability among monkeys derived from different countries has not been published, especially on the key enzyme, cytochrome P450 (CYP450). We assessed the amounts of hepatic CYP450 obtained from cynomolgus monkeys of different ages and from different countries in this study. There were no regional differences of total P450 content, as well as major CYP450 isozymes (CYP 1A, 2A, 2B, 2C, 2D, 2E1 and 3A4) in cynomolgus monkeys by westernblot analysis. Similarly, there were no significant differences with hybrid cynomolgus monkeys, but variations in individual values were large. As for aging, total P450 contents declined in old cynomolgus monkeys (12-32 years of age). These results indicate the usefulness of basic data of hepatic CYP450 obtained from cynomolgus monkeys of different ages and from different countries. PMID- 15897622 TI - Estrous stage- and animal age-independent superovulation in the BrlHan:WIST@Jcl(GALAS) rat. AB - In most strains of rats, the effects of treatment for the induction of superovulation show major strain differences and are strongly influenced by the stage of the estrous cycle. This study demonstrated, however, that superovulation was easily induced in Wistar strain Brl Han:WIST@Jcl(GALAS) rats by PMSG and hCG administration. To confirm the effects of such treatment, we studied age differences in egg collection efficiency. After superovulation was induced by intraperitoneal administration of 150 IU/kg PMSG and 75 IU/kg hCG given 48 h apart, the mean numbers of oocytes obtained from rats at 4, 8, 12, 20 and 28 weeks of age were 38.9, 33.5, 46.1, 26.9 and 21.3, respectively. No differences caused by the estrous stage at the PMSG administration were observed. In an embryo transfer experiment, fertilized eggs obtained from superovulated rats at each week of age showed equivalent viability until full-term to those from untreated rats. These results suggest that estrous stage-independent superovulation is effective not only in the pre-pubertal stage but also in adult rats. PMID- 15897623 TI - Characteristic intestinal microflora of specific pathogen-free mice bred in two different colonies and their influence on postnatal murine immunocyte profiles. AB - Cecal microflora of BALB/c mice originating from two different SPF-breeding colonies were compared. The analysis of cultivable bacteria in the ceca showed significantly higher numbers of total bacteria in BALB/cCrSlc (SLC mice) than in BALB/cA Jcl (JCL mice) (p<0.05), which were mainly based on higher numbers and occurrence of Peptococaceae. Bifidobacteria were detected only in SLC mice. Feeding an oligosaccharide, raffinose, to the mice also induced different shifts in the composition of cecal microflora and the concentration of cecal organic acids. In the second experiment, hysterectomy-derived (HD) SLC mice were fostered to SPF lactating SLC mothers, or SPF lactating JCL mice, together with the mother's own natural birth (NB) pups in each isolator. HD mice fostered to SLC mothers showed significantly higher percentages of T-cell receptor alphabeta cells expressing a CD8alpha homodimer (p<0.05) and a CD8alphabeta heterodimer (p<0.001) in the intraepithelial lymphocytes (IEL) compared with HD mice fostered to JCL-mothers. IEL profiles of HD mice corresponded well to those of NB mice that were breast-fed by the same mothers. Differences in the ratio of B220(+)cells to Thy1.2(+)cells in the splenocytes were also observed as a trend between both HD mice fostered to SLC or JCL mothers (p=0.06). These results suggest that postnatal colonization of various characteristic intestinal microflora derived from SPF-breeding colonies results in differences in development of lymphocyte populations in the intestinal and systemic organs of mice. PMID- 15897624 TI - Effects of housing conditions on the development of wet skin lesions in the NOA mouse. AB - The effects of housing on the onset time and prevalence of wet skin lesions were investigated in NOA mice, which spontaneously develop these lesions at a high rate. Wet skin lesions developed earliest in mice that were housed individually. For mice that were housed in groups, the lesions developed earlier in mice with non-littermate group housing than in mice with littermate group housing. The prevalence of lesions was in the following order: individual housing > non littermate group housing > littermate group housing. These results suggest that socio-psychological factors are involved in the etiology of wet skin lesions in the NOA mouse. Under individual housing conditions, two other novel characters of the NOA mouse were also observed, specifically, development of dry skin and wet skin lesions at the tail root. These characteristics developed early and with high prevalence and were easily observed on external examination. Therefore, these novel characteristics observed in NOA mice are potential markers of the psychological state of the animals. PMID- 15897625 TI - Viability and plasma vitamin K levels in the common bile duct-ligated rats. AB - The common bile duct-ligated (CBDL) rat, which is widely used as a model of human cirrhosis, rapidly develops secondary biliary cirrhosis (SBC) within 4 weeks. The CBDL rat shows poor viability, however, a detailed examination of the causes of its death has not been made. In this study, we investigated the outcome of bile duct ligation in detail and attempted to extend the life span of this model by feeding the animals a diet supplemented with nutrients. Survival rate, blood chemistry, blood cell counts, plasma levels of K vitamins and liver histology were compared among CBDL rats fed a standard diet and an enriched diet. Sham operated rats were used as a control. Six out of 18 CBDL rats fed the standard diet died within 32 days of operation. The cause of death was massive internal hemorrhage in various organs or body cavities. All CBDL rats fed the enriched diet survived more than 31 days, but the viability of CBDL rats was not significant between those fed the standard diet and the enriched diet. The degree of anemia correlated significantly with the prolongation of prothrombin time. Plasma vitamin K1 levels in CBDL rats were significantly lower than those in sham operated rats, but vitamin K2 levels were similar. We suggest that massive hemorrhage, which was the direct cause of death, is caused by the impairment of hemostasis resulting from vitamin K deficiency. The enriched diet with vitamin K nutritional supplements seemed to contribute to the prolongation of the life span of CBDL rats. PMID- 15897626 TI - Newly designed primer sets available for evaluating various cytokines and iNOS mRNA expression in guinea pig lung tissues by RT-PCR. AB - Guinea pigs are often used as an animal model of human tuberculosis (TB). However, there are few methods available for pursuing the immunological processes involved in guinea pig TB. In this study, we developed for the first time systematic reverse transcription (RT)-PCR for evaluation of guinea pig mRNA expression. RT-PCR primer sets were newly designed for detection of cytokines and inducible nitric oxide synthase (iNOS) mRNA in guinea pig TB. Interferon (IFN) gamma, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, interleukin (IL)-1beta, IL-2, IL-10, IL-12p40, granulocyte-macrophage-colony stimulating factor (GM-CSF) and iNOS mRNA expression were detected significantly and reproducibly when these primer sets were used. The data by real-time PCR were comparable with those of RT-PCR. We showed that these RT-PCR primer sets could be used to examine mRNA expression semi-quantitatively in guinea pig tissues, and conclude that these newly designed primer sets for conventional RT-PCR will be useful for studying the immunological processes in guinea pig tuberculosis experiments to investigate and evaluate efficacy of new vaccines or anti mycobacterial drugs. PMID- 15897627 TI - Development and characterization of CATS markers for genetic linkage mapping in the house musk shrew, Suncus murinus. AB - To serve as an initial step in developing an ideal genetic marker map for the house musk shrew, Suncus murinus, 318 comparative anchor tagged sequence (CATS) primer pairs were assessed for polymorphism ascertainment and linkage mapping. Of the 112 (35.2%) CATS primer pairs that were successfully amplified by PCR in the shrew, 18 (16.1%) showed polymorphism between two mutant strains, BAN-kc, oeb and WZ. Linkage analysis of the polymorphic CATS markers and three visible mutant genes, kc, oeb and wz, genotyped in a 77 F2 mapping panel from a cross of the two mutant strains, assigned wz and five CATS markers into three linkage groups. Sequence analysis revealed that two (ADA and TXN) out of nine CATS amplified sequences had a total of six deletions of varying sizes and 17 single nucleotide polymorphisms (SNPs). BLAST search identified three CATS (ADA, CYP1A2, and TXN) products matching the genes from which they were originally designed, while the remaining six markers could not be identified. Together with the use of the detected SNPs as genetic markers, the five CATS markers linkage mapped in this species will serve as anchors in establishing the first framework map for locating loci affecting all heritable qualitative and quantitative traits in the musk shrew. PMID- 15897628 TI - Effect of human erythropoietin (hEPO) treatment on anemia in ICR-derived glomerulonephritis (ICGN) mice. AB - ICR-derived glomerulonephritis (ICGN) mice are a novel inbred strain with hereditary nephrotic syndrome and are thus considered a good animal model of human idiopathic nephrotic syndrome. In the present study, we investigated the effect to erythrocyte production by human erythropoietin (hEPO) treatment in ICGN mice during the early nephrotic stage. Erythrocyte count, hemoglobin concentration and hematocrit value in hEPO-treated (5 U/body/day, for 5 days) ICGN mice were recovered to the levels found in normal ICR mice. In addition, there was no correlation between plasma creatinine level, a marker of renal function, and erythrocyte count after hEPO treatment. Therefore, anemia in ICGN mice may be caused by decreased production of EPO in the kidney following progressive parenchymal damage. PMID- 15897629 TI - Estrous cycle stage-independent treatment of PMSG and hCG can induce superovulation in adult Wistar-Imamichi rats. AB - The estrous cycle influence on the number of ovulated eggs after injection of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) was investigated in 12, 18, and 24 weeks old adult female Wistar-Imamichi (WI) rats. PMSG (150 IU/kg) was injected at metestrus, diestrus, proestrus, or estrus, followed by hCG (75 IU/kg) 55 h later. Ovulation was induced at all ages and stages of the estrous cycle. The number of ovulated eggs was not affected by stage for similarly aged rats, however, the number of ovulated eggs obtained after treatment decreased with age. These results demonstrate that the PMSG/hCG treatment can induce ovulation at any stage of estrous cycle in WI rats and efficient superovulation at 12 weeks of age. PMID- 15897630 TI - Battery of tests for profiling abnormalities of vitamin K-dependent coagulation factors in drug-toxicity studies in rats. AB - A battery of simple tests for profiling abnormalities of vitamin K-dependent coagulation factors encountered in drug-toxicity studies was verified in rats treated with warfarin (3 and 10 mg/kg, p.o). The thrombotest, or hepaplastin test, is useful as a follow-up test after routine screening tests for coagulation abnormalities based on PT and APTT, to rule out other coagulation-factor abnormalities. Measurement of coagulation factor activities (factors II, VII, IX and X) using factor-deficient human plasmas provides direct evidence of decreased activities of vitamin K-dependent factors. Furthermore, Echis carinatus venom coagulation time, together with factor II activity, allows us to confirm the generation of PIVKA-II. PMID- 15897631 TI - Transcutaneous cisternal puncture for sampling of cerebrospinal fluid in awake rat. AB - Reported cisternal puncture methods require the anesthetization and fixation of an animal within a stereotaxic frame. To determine the effect of anesthesia and animal fixation on the central nervous system (CNS), amino acid concentrations of cerebrospinal fluid (CSF) sampled by transcutaneous cisternal puncture were compared among awake rats, pentobarbital-anesthetized rats and pentobarbital anesthetized rats fixed in a stereotaxic frame. Although the concentrations of many amino acids in the CSF of pentobarbital-anesthetized rats were lower than in awake rats, use of the stereotaxic frame resulted in significantly increased amino acid concentrations in the CSF. These data indicate that CSF sampling by transcutaneous cisternal puncture from awake rats is a suitable method for serial measurement of drug effects on the CNS. PMID- 15897632 TI - Recent advances in the pathophysiology and clinical management of anemia. PMID- 15897633 TI - MIDD and MELAS: a clinical spectrum. PMID- 15897634 TI - Hemophagocytic syndrome. PMID- 15897635 TI - Nocturnal polyuria with abnormal circadian rhythm of plasma arginine vasopressin in post-stroke patients. AB - OBJECTIVE: Nocturia is a common reason for interrupted sleep in post-stroke patients. These patients often have neurogenic bladder overactivity. However, little is known about the possible contribution of nocturnal polyuria in the patients. METHODS: We measured the number of nocturia, the circadian plasma arginine vasopressin (AVP) level and urinary excretion in 4 patients with stroke. RESULTS: All patients had nocturnal urinary frequency (three times in one and twice in 3). All patients were revealed to have nocturnal polyuria, and the ratio of nocturnal urinary output to 24 hour volume ranged from 36% to 63%. Measurement of daily plasma AVP variation showed that all patients lost normal nocturnal rise of the plasma AVP concentration. Two patients were successfully treated with 5 mug of intranasal desmopressin once a night, a potent analogue of AVP, without hypertension particularly in the night, signs of congestive cardiac failure or any electrolyte abnormality such as hyponatremia. CONCLUSION: Our post-stroke patients had nocturnal polyuria with abnormal circadian rhythm of plasma AVP secretion. Desmopressin reduced nocturnal waking in urination. It also ameliorated nocturnal dehydration that might trigger a stroke recurrence in the patients. PMID- 15897636 TI - Hemochromatosis with mutation of the ferroportin 1 (IREG1) gene. AB - The HFE, H ferritin, TFR2, and ferroportin 1 genes of a Japanese patient diagnosed as having hemochromatosis were amplified by PCR and sequenced. A novel mutation in the ferroportin 1 was found in the patient. It was located in the noncoding region of the ferroportin 1; nucleotide 117 adenine was changed to guanine, 7 nucleotides downstream the iron responsive element (IRE) region. This mutation was not found in the patient's son or daughter, or in 50 healthy individuals. It was suggested that the mutation in the ferroportin 1 may be related to hemochromatosis of this patient. PMID- 15897637 TI - Hemoperitoneum secondary to spontaneous rupture of hepatic metastasis from lung cancer. AB - We describe a rare case of hemoperitoneum secondary to spontaneous rupture of hepatic metastasis from lung cancer. A 72-year-old man with non-small cell lung cancer was admitted to our hospital with sudden onset of right upper abdominal pain and hypovolemic shock. Laboratory tests showed severe anemia. Abdominal contrast-enhanced computed tomography revealed massive ascites and multiple liver metastases. Rupture of a metastatic liver tumor was suspected. Only palliative therapy was performed. The patient's general condition gradually worsened, and he died 2 months after admission. Autopsy examination revealed hemoperitoneum due to a ruptured metastatic liver tumor originating from pulmonary squamous cell carcinoma. PMID- 15897638 TI - Alpha-fetoprotein (AFP)- and des-gamma-carboxy prothrombin (DCP)-producing adenocarcinoma of the stomach with liver metastasis in a patient with chronic hepatitis C. AB - A 45-year-old man was admitted to our hospital because of chronic hepatitis C and a large liver tumor accompanied by increased serum levels of alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP), the tumor markers for hepatocellular carcinoma. Endoscopic examination revealed advanced gastric cancer. Biopsy specimens of the stomach and liver showed gastric adenocarcinoma and its metastasis to the liver. Immunohistochemical studies demonstrated that adenocarcinoma cells both of the stomach and liver, were positive for the antibodies against AFP as well as DCP. Expression of AFP mRNA was shown in the cancer cells of the stomach. Accordingly, we diagnosed this patient with AFP- and DCP-producing adenocarcinoma of the stomach together with liver metastasis. PMID- 15897639 TI - Seroconversion of glutamic acid decarboxylase antibodies in a patient initially diagnosed as having type 2 diabetes mellitus. AB - A 61-year-old man admitted in July 1998 had suffered from thirst, polydipsia and polyuria for three years. Diet and transient insulin therapy had induced good blood glucose control which was maintained by metformin hydrochloride for a year. Although it worsened, conventional insulin treatment re-implemented good blood glucose control. Glutamic acid decarboxylase antibodies (GAD-Ab) had been negative up to this point. After 8 months, blood glucose levels became elevated. To date, the GAD-Ab has been positive (112-120 U/ml), and the serum and urine C peptide levels are decreased. Seroconversion of GAD-Ab should be noted in patients initially diagnosed as having GAD-Ab negative type 2 diabetes mellitus. PMID- 15897640 TI - Autoimmune thyroid disease (Graves' disease and hashimoto's thyroiditis) in two patients with Crohn's disease: case reports and literature review. AB - An increased prevalence of the association between autoimmune thyroid diseases and ulcerative colitis has been suggested, however, not with Crohn's disease, as only 7 cases of thyroid disease coexisting with Crohn's disease have been reported. Herein, we describe 2 patients with Crohn's disease complicated with Graves' disease or autoimmune thyroiditis, and also review other cases with those complications. Some immunological processes are suggested to be implicated in the pathogenesis of this association, however, the exact mechanism remains unclear. PMID- 15897641 TI - Hypercalcemia in a patient with primary hyperparathyroidism and acromegaly: distinct roles of growth hormone and parathyroid hormone in the development of hypercalcemia. AB - We herein report a case of primary hyperparathyroidism associated with acromegaly. Although serum parathyroid hormone (PTH) levels increased after the resection of a pituitary adenoma, levels of serum 1a, 25-dihydroxyvitamin D [1, 25(OH)2D] decreased but remained above the normal upper limit. After resection of a parathyroid adenoma, serum PTH, 1, 25(OH)2D, calcium (Ca), and phosphate were all normalized. Since serum 1, 25(OH)2D levels decreased in spite of the increase in serum PTH levels after normalization of levels of growth hormone (GH), GH may have contributed to the elevation of serum 1, 25(OH)2D. It is therefore suggested that the mechanism by which elevation of serum 1, 25(OH)2D occurred in the present case may involve an increase in serum GH distinct from the PTH-mediated pathway. PMID- 15897642 TI - Autosomal dominant polycystic kidney disease showing rupture of a lateral ventral hernia following paralytic ileus. AB - We report an 83-year-old Japanese male with autosomal dominant polycystic kidney disease (ADPKD), which was marked by unusually enlarged kidneys, and in whom ileus occurred after administration of procainamide. The bowels became swollen and ruptured the skin and appeared on the outside of the skin. Even after the ileus state was resolved, the projected intestinal tract was not restored due to a large defect of the skin, and ostomy was performed. Abdominal hernia including lateral ventral hernia due to enlarged kidneys may result in perforation of the abdominal wall as well as intestinal wall. PMID- 15897643 TI - Small cell lung cancer associated with systemic sclerosis. AB - A 51-year-old woman developed small cell lung cancer (SCLC). She was a non-smoker and had interstitial pneumonia associated with systemic sclerosis (SSc). Sixteen additional cases obtained from the literature describing patients with SCLC associated with SSc are reviewed. The majority of patients were women with underlying interstitial pneumonia. In 3 patients who were non-smokers, interstitial pneumonia was complicated and cancer had developed in the peripheral lung field but not in the central lung field. Since SCLC is very rare in non smokers, these findings suggest a positive association between SCLC and interstitial pneumonia associated with SSc. PMID- 15897644 TI - Long-term clinical course of a patient with anti PL-12 antibody accompanied by interstitial pneumonia and severe pulmonary hypertension. AB - We report a case of a patient with anti PL-12 antibody accompanied by interstitial pneumonia and severe pulmonary hypertension. At first presentation, hyperkeratotic skin lesions were found, although the diagnosis of CVD was not conclusive. Lung histology showed diffuse fibrosing interstitial pneumonia predominantly in the subpleural regions. During the seven-year follow-up period, severe pulmonary hypertension developed, although the progression of lung fibrosis was relatively limited. Anti-PL12 antibody was detected, and therefore the patient was diagnosed as having antisynthetase syndrome. Lung histology and pulmonary arteriogram suggested that vascular involvement of the disease contributed to the development of severe pulmonary hypertension. PMID- 15897645 TI - Embolic stroke with a patent foramen ovale and Behcet's disease. AB - We report a case of a 53-year-old man with Behcet's disease (BD) and paradoxical stroke. This patient suddenly developed sensory aphasia, right oculomotor palsy, nystagmus, and left ataxic hemiparesis. MR images showed fresh embolic infarcts in the midbrain and the left parietotemporal lobe. He presented with uveitis, oral aphthous ulcers and genital ulcers. An electrocardiogram revealed no abnormalities, and a transesophageal echocardiogram disclosed a patent foramen ovale. Since venous involvement is common in BD, we suspected the presence of smaller thrombi probable in this patient. We conclude that embolic stroke is a notable complication of BD with right-to-left shunting. PMID- 15897646 TI - A mother and a child with maternally inherited diabetes and deafness (MIDD) showing atrophy of the cerebrum, cerebellum and brainstem on magnetic resonance imaging (MRI). AB - A mother and a child with maternally inherited diabetes and deafness (MIDD) showing atrophy of the cerebrum, cerebellum and brainstem on magnetic resonance imaging (MRI) were reported. The proband had slowly progressive cerebellar ataxia and her son had depression. Mitochondrial DNA purified from their leucocytes had the heteroplasmic point mutation at position 3243 (A-->G). Involvement of the central nervous system should be considered in MIDD as well as in other mitochondrial diseases. PMID- 15897647 TI - Spontaneous haemophilus influenzae type B meningoventriculitis with intraventricular debris. AB - Here we report the first case of spontaneous occurrence of Haemophilus influenzae type B meningoventriculitis. A 47-year-old man suffered from fever and headache. He had neck stiffness, and his cerebrospinal fluid (CSF) was turbid and yellowish. Gram staining and latex agglutination test of his CSF indicated that Haemophilus influenzae type B was the causative organism. Cranial CT and MRI revealed right ventricular debris accumulation, periventricular hyperintense signal, and periventricular ependymal enhancement. He was treated with intravenous administration of antibiotics, and his clinical condition and MRI findings gradually improved. Six weeks after onset, he was discharged without sequelae. PMID- 15897648 TI - Felty's syndrome with chronic hepatitis and compatible autoimmune hepatitis: a case presentation. AB - A 68-year-old woman with Felty's syndrome was admitted to our hospital due to breathlessness. She was diagnosed as having rheumatoid arthritis at age 59 years. Abdominal computed tomography indicated ascites, splenomegaly and liver atrophy. She had no antigens or antibodies for hepatitis virus, or antibodies for mitochondria with the exception of antinuclear antibody. According to the International Autoimmune Hepatitis (AIH) scoring system, she was diagnosed as having chronic hepatitis, compatible with AIH. The association of Felty's syndrome with AIH is very rare and the most difficult problem to overcome is whether or not steroid therapy is necessary in patients with Felty's syndrome complicated by AIH. PMID- 15897649 TI - Virus-associated hemophagocytic syndrome in an international traveler as a differential diagnosis of SARS. AB - During the epidemic of severe acute respiratory syndrome in 2003, a 27-year- old Japanese woman presented a high fever and acute respiratory distress with pulmonary infiltrates after traveling to a high-risk area. An alternative diagnosis was made as Epstein-Barr virus-associated hemophagocytic syndrome, based on the proliferation of macrophages with hemophagocytosis in the bone marrow and Epstein-Barr viral marker profiles. Virus-associated hemophagocytic syndrome in an international traveler should be included in the differential diagnosis of severe acute respiratory syndrome. PMID- 15897650 TI - Retropharyngeal abscess due to methicillin-resistant Staphylococcus aureus in a case of acute myeloid leukemia. AB - We describe a case of acute myeloid leukemia (AML) complicated with retropharyngeal abscess (RPA) due to methicillin-resistant Staphylococcus aureus (MRSA) in a 56-year-old man. After administration of vancomycin and lavage of the retropharyngeal space with gentamicin, complete resolution of the RPA was observed. Despite their lower frequency, deep neck infections are associated with high mortality rates. The possibility of RPA should be considered in patients who present with fever, dysphagia and limitation of neck extension. Lavage of the retropharyngeal abscess with gentamicin may be optimal in cases of large RPA. PMID- 15897651 TI - Lemierre's syndrome: Porphyromonas asaccharolytica as a putative pathogen. AB - A case of Lemierre's syndrome is reported in which metastatic abscesses resulted from septic thrombophlebitis of the internal jugular vein secondary to bacterial pharyngitis. A 32-year-old male suffering from a painful left-sided neck mass, sore throat, and fever was admitted to our hospital. Computed tomography revealed thrombosis of the left internal jugular vein, septic pulmonary emboli, and a liver abscess. Blood culture showed Porphyromonas asaccharolytica. Although empyema occurred transiently during the treatment, the patient recovered following prolonged antimicrobial therapy. Although Fusobacterium species are a well-known cause of Lemierre's syndrome, cases in whom Porphyromonas species was isolated have scarcely been reported. Moreover, case reports from Japan have been few. PMID- 15897652 TI - Impact of sex and age on coronary basal tone. PMID- 15897653 TI - Asymptomatic double aortic arch. PMID- 15897654 TI - A spinal dural arteriovenous fistula with lower motor neuron involvement. PMID- 15897655 TI - Therapeutic strategy for acute stroke--prologue for an epoch of brain attack--. Progress of diagnosis of acute stroke. PMID- 15897656 TI - Therapeutic strategy for acute stroke--prologue for an epoch of brain attack--. Acute stroke therapy based on Japanese guidelines for the management of stroke 2004. PMID- 15897657 TI - Therapeutic strategy for acute stroke--prologue for an epoch of brain attack--. The present state of acute ischemic stroke therapy in Japan. PMID- 15897658 TI - Therapeutic strategy for acute stroke--prologue for an epoch of brain attack--. EBM and current state in Japan of thrombolytic therapy for acute ischemic stroke. PMID- 15897659 TI - Therapeutic strategy for acute stroke--prologue for an epoch of brain attack--. Future aspects of gene therapy in acute ischemic stroke. PMID- 15897660 TI - Appropriate use of antibiotics and guideline(s) for this purpose. Present situation of antimicrobial resistance in Japan. PMID- 15897661 TI - Appropriate use of antibacterial drugs and guidelines: special aspects of Japan's guidelines. PMID- 15897662 TI - Appropriate use of antibiotics and guideline(s) for this purpose. Outcome of the antimicrobial drug guideline. PMID- 15897663 TI - Appropriate use of antibiotics and guideline(s) for this purpose. The appropriate use of antimicrobial agents in the treatment of infectious diseases and guidelines future trends, "global standard ?" or "regional standard ?". PMID- 15897664 TI - New aspects of renal diseases--from urine checkup to kidney diseases--. Renal screening tests. PMID- 15897665 TI - New aspects of renal diseases--from urine checkup to kidney diseases--. Importance of early detection and diagnosis for kidney diseases. PMID- 15897666 TI - New aspects of renal diseases--from urine checkup to kidney diseases--. Management and treatment of patients with chronic kidney disease. PMID- 15897667 TI - New aspects of renal diseases--from urine checkup to kidney diseases--. Comprehensive patient care system from medical checkups for kidney diseases to kidney transplantation-dialysis therapy. PMID- 15897668 TI - Markers of endothelial cell activation/injury: CD146 and thrombomodulin are related to adiponectin in kidney allograft recipients. AB - BACKGROUND: Adiponectin may be used for assessing the risk of coronary artery disease (CAD) and may be related to the development of acute coronary syndrome. Decreased adiponectin has been associated with some risk factors for cardiovascular diseases such as male sex, obesity and diabetes mellitus. Adiponectin has antiatherogenic properties and attenuates endothelial inflammatory responses. CD146, a novel cell adhesion molecule, is localized at the endothelial junction. In kidney allograft recipients, endothelial dysfunction and atherosclerosis are almost universal. The aim of this cross-sectional study was to evaluate possible relations between adiponectin, CD146, and other markers of endothelial cell injury in 82 stable kidney transplant recipients (mean age 45 years, mean time after transplantation 47 months) with and without CAD. METHODS: Adiponectin and markers of endothelial injury: CD146, von Willebrand factor, thrombomodulin, ICAM, CD40L, P-selectin and other hemostatic markers were assessed using commercially available kits. RESULTS: Patients with CAD had evidence of more pronounced endothelial dysfunction, procoagulant state and lower adiponectin than patients without CAD. Adiponectin correlated significantly, in univariate analysis, with CD146 (r = 0.29, p = 0.009), thrombomodulin (r = 0.37, p = 0.001), protein Z (r = -0.25, p = 0.03), BMI (r = -0.26, p = 0.047), serum creatinine (r = 0.26, p = 0.02) and urea (r = 0.38, p = 0.001). CD146 correlated significantly with von Willebrand factor (r = 0.33, p = 0.002), thrombomodulin (r = 0.25, p = 0.025), age (r = 0.34, p = 0.001), platelets (r = -0.33, p = 0.002), serum urea (r = 0.24, p = 0.039), cholesterol (r = 0.24, p = 0.046), ICAM (r = 0.23, p = 0.036), protein C activity (r = -0.26, p = 0.019) and tended to correlate with serum creatinine and time after transplantation. In multivariate linear regression, independent predictors of adiponectin were CD146, thrombomodulin and urea, and of CD146 was mainly age of patients. CONCLUSIONS: Endothelial dysfunction and procoagulant state are more pronounced in kidney transplant recipients with CAD, particularly in those with lower GFR. In kidney transplant recipients, markers of endothelial cell injury are significantly increased relative to healthy volunteers. Elevation of adiponectin may be a defense mechanism against endothelial damage, reflected by elevated CD146 and thrombomodulin. PMID- 15897669 TI - Ovalbumin-induced IL-4, IL-5 and IFN-gamma production in infants with atopic dermatitis. AB - BACKGROUND: Although hen's eggs are considered a cause of infantile atopic dermatitis (AD), little is known about cytokine production upon egg stimulation in infants with AD. OBJECTIVE: This study aimed to characterize the production of IL-4, IL-5 and IFN-gamma upon stimulation with ovalbumin (OVA), a representative allergenic protein of egg, in infants with AD. METHODS: Peripheral blood mononuclear cells (PBMCs) from 68 children with AD, including 46 infants (<1 year), were stimulated with OVA and the production of IL-4, IL-5 and IFN-gamma was measured with ELISA kits. RESULTS: Upon stimulation with OVA, the production of IL-4 and IL-5, but not IFN-gamma, by PBMCs was significantly higher in infants with AD than in non-atopic controls. OVA-induced IL-5 production peaked in younger infants (2-5 months) and then decreased with age increase. In contrast, OVA-induced IL-4 production peaked at the age of 1-2. This coincided with the serum level of egg white-specific IgE (EW-IgE). There was a significant positive correlation between IL-5 production and the severity of symptoms in infants with AD, while IL-4 production significantly correlated with the serum level of EW IgE. CONCLUSION: These results demonstrate that OVA-induced IL-5 production fluctuates with age in a different manner than IL-4 or EW-IgE. Our results suggest that egg contributes to the development of AD in younger infants by inducing the production of IL-5, but not IL-4. PMID- 15897670 TI - Human hair: an unexpected source of cat allergen exposure. AB - BACKGROUND: Cat allergens are ubiquitous because the clothing of cat owners constitutes an important source of distribution of Fel d 1 in cat-free environments. Since Fel d 1 can adhere to a variety of surfaces, we sought to verify if human hair belonging to individuals with or without a cat at home might represent a reservoir and be a possible carrier of cat allergens. METHODS: Seventy-three women (25 with a non-neutered male cat and 25 with a dog at home, and 23 controls without any direct contact with these animals) were recruited. The collection of material from hair was carried out using a modified version of a battery-powdered portable sampler. Particulate material was harvested onto glass fiber filters (25 mm in diameter, with a pore size of 2 microm; AP 20 Millipore, Milan Italy), extracted in phosphate buffer with BSA and then assayed for the evaluation of cat allergen using an ELISA based on anti-Fel d 1 monoclonal antibody. RESULTS: Detectable levels of cat allergen were found in 2 controls, in 2 women with a dog at home and in 13 women with a cat at home, respectively. CONCLUSIONS: In some women with a cat at home, hair constitutes a significant reservoir of Fel d 1. It is likely that these amounts of cat allergen might contribute to allergic sensitization when released in cat-free environments. PMID- 15897671 TI - Celecoxib tolerability in patients with hypersensitivity (mainly cutaneous reactions) to nonsteroidal anti-inflammatory drugs. AB - BACKGROUND: Adverse reactions to nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly observed in clinical practice, particularly among patients with chronic urticaria or asthma. The identification of a safe and reliable alternative is a frequent problem for both general practitioners and allergists. METHODS: We assessed 120 patients (83 women and 37 men) who had experienced adverse reactions to one or more NSAIDs; 64 (53.3%) of them had reacted to only one NSAID (single reactors) and 56 (46.7%) to multiple NSAIDs (multiple reactors). Among our subjects, 76.7% reported cutaneous reactions, 8.3% respiratory symptoms, 10.8% both cutaneous and respiratory symptoms, and 4.2% anaphylaxis. All patients were subjected to a single-blind, placebo-controlled oral challenge with two different doses of celecoxib (50 + 150 mg 1 h later = cumulative dose of 200 mg). RESULTS: None of the patients reacted to the placebo and only one (0.8%) suffered a reaction (urticaria) after the second dose of celecoxib. CONCLUSIONS: Celecoxib showed a 98.9% rate of tolerability in the 92 patients with exclusively cutaneous reactions and was well tolerated by all 28 subjects with NSAID-related respiratory or anaphylactic symptoms. PMID- 15897672 TI - No association between the ionotropic glutamate receptor kainate 3 gene ser310ala polymorphism and schizophrenia. AB - The etiology of schizophrenia has been suggested to be associated with the dysfunction of the glutamatergic system. A positive association between the ionotropic glutamate receptor kainate 3 gene (GRIK3) T928G polymorphism and schizophrenia has been reported, which suggests that people bearing G allele are at a higher risk for schizophrenia. Therefore, we attempted to replicate this study in Chinese with larger sample size. One hundred and sixty schizophrenic patients and 160 healthy controls participated in the current study. Genotype and allele distributions of GRIK3 T928G polymorphism in schizophrenics were similar to those of controls (p = 0.74 and p = 0.59, respectively). Our study indicated that there was no association between this polymorphism and schizophrenia, at least in Chinese. PMID- 15897673 TI - Effects of different subanesthetic doses of (S)-ketamine on neuropsychology, psychopathology, and state of consciousness in man. AB - This is the first neuropsychological study using the S-enantiomer of the noncompetitive N-methyl-D-aspartate antagonist ketamine. In 2 randomized placebo controlled trials we studied effects of two different doses of (S)-ketamine (low dose/high dose) on neuropsychological functions and psychopathology in 12 healthy male volunteers. Impairment was measured via standardized neuropsychological tests. Results indicate that both subanaesthetic doses produce only nonsignificant impairment in most of the tasks. Tasks involving divided and sustained attention as well as scores for objective and subjective psychopathology show significant impairment in a dose-dependent manner. Implications of these findings for the neuropsychology of attention and schizophrenia are discussed. PMID- 15897674 TI - Partial normalization of serum brain-derived neurotrophic factor in remitted patients after a major depressive episode. AB - We had previously reported decreased serum brain-derived neurotrophic factor (BDNF) levels in depressed patients. In the present study, we tested the hypothesis that antidepressant treatment would normalize serum BDNF levels, at least in a subgroup of patients. Major depressed patients (15 females and 11 males) diagnosed according to DSM-IV criteria and healthy controls (13 females and 13 males) participated in this study. Serum BDNF was assayed with the ELISA method for depressed and remitted patients and the severity of depression was evaluated with the Montgomery-Asberg Depression Rating Scale. An analysis of variance showed that treatment had an effect [F(1, 24) = 4.46, p = 0.045] on the normalization of serum BDNF levels. We also found a correlation between the severity of depression (r = 0.51, p = 0.008), the pretreatment BDNF levels (r = 0.62, p = 0.001) and the difference in serum BDNF levels after antidepressant treatment. These results suggest that antidepressant treatment has a positive effect on serum BDNF levels and support the hypothesis of neurotrophic factor involvement in affective disorders. PMID- 15897675 TI - A possible antienuretic effect of reboxetine in children and adolescents with attention deficit/hyperactivity disorder: case series. AB - Reboxetine is a selective norepinephrine reuptake inhibitor. OBJECTIVES: To evaluate the antienuretic effect of reboxetine in children with attention deficit and mood disorders. METHODS: Six children aged 12-15 years with primary nocturnal enuresis and with attention deficit/hyperactivity disorder with or without other comorbid disorders were treated with reboxetine (4-8 mg/day). Monitoring of enuretic frequency was recorded at baseline and once a week for 6 weeks. RESULTS: A statistically significant decrease in bedwetting was noted along the assessment points (p < 0.001), with the maximal change noted in the first week of treatment (p = 0.004). Reboxetine was well tolerated with minimal and transient side effects. CONCLUSIONS: reboxetine may possess an antienuretic effect in some children with the attention deficit/hyperactivity disorder. PMID- 15897676 TI - Unexpected effects of nitric oxide synthase inhibitors on extracellular nitrite levels in the hippocampus in vivo. AB - The aim of this study was to determine whether extracellular nitric oxide levels in the hippocampus of freely moving animals were reduced by the administration of nitric oxide synthase (NOS) inhibitors via a microdialysis probe. Our results show that extracellular nitrite levels were increased following the infusion of N nitro-L-arginine methyl ester (L-NAME) and 7-nitroindazole (7-NI), in the case of the latter, the response was biphasic. In contrast, infusion of both inhibitors together resulted in a substantial reduction in nitrite when compared to control. More predictably, the infusion of NMDA elicited an increase in extracellular nitrite levels. This effect was biphasic, the second phase occurring some 3 h after the drug infusion period had ended. When NMDA was infused in the presence of L-NAME, no agonist-induced increase in nitrite production was recorded, in fact nitrite levels were found to decline to below control values. There was no immediate increase in nitrite levels when NMDA was infused in the presence of 7 NI, although this may have been partially obscured by the biphasic effect of the inhibitor. It did appear, however, that the second phase of the NMDA-induced response was attenuated by 7-NI. No NMDA-evoked increase in nitrite was evident when the agonist was infused in the presence of a combination of both inhibitors. We have no clear explanation for the data presented here but suggest that up regulated activity of particular NOS isoforms might compensate for the inhibition of the other by a mechanism yet to be elucidated. In addition, we propose that caution be exercised when interpreting results from in vivo microdialysis studies where NOS inhibitors are administered directly into the brain via a probe. PMID- 15897677 TI - Effect of fentanyl on lidocaine-induced convulsions in mice. AB - This study was undertaken to examine the effect of fentanyl on lidocaine-induced convulsions in mice. Seventy-five male mice were used and divided into five groups, 15 in each. Convulsions were obtained by lidocaine injection subcutaneously (150 mg/kg), in a dose volume of 0.1 ml/10 g body weight over the upper back. The five groups were: (a) control group: pretreated with normal saline; (b) F50 group: pretreated with fentanyl 50 microg/kg; (c) F100 group: pretreated with fentanyl 100 microg/kg; (d) F200 group: pretreated with fentanyl 200 microg/kg, and (e) F100+N group: pretreated with fentanyl 100 mug/kg plus naloxone 1 mg/kg. The pretreatments were given intraperitoneally (i.p.) 5 min prior to lidocaine injection. After lidocaine injection, the latency to the onset of generalized convulsions was observed for 12 min and recorded. The severity of convulsions was assessed and scored as 1 = mild, 2 = moderate, and 3 = severe. The recovery or death of the mice were also recorded. Student's t, Mann-Whitney U, and Fisher's exact tests were used to analyze the data. Compared with the control group (431.7 +/- 37.3 s), the latencies of onset of convulsions in the fentanyl groups were dose-dependently decreased (F50: 331.0 +/- 37.1 s; F100: 240.3 +/- 28.6 s, p < 0.001; F200: 188.7 +/- 19.4 s, p < 0.001), and the decreased latency was reversed by naloxone (F100+N: 412.9 +/- 34.1 s). Compared with the control group (1.13 +/- 0.19), the severities of convulsions in the fentanyl groups were also increased in a dose-dependent manner (F50: 1.47 +/- 0.19; F100: 1.93 +/- 0.21, p < 0.05; F200: 2.46 +/- 0.17, p < 0.001). Similarly, the increased severity was reversed by naloxone (F100+N: 1.33 +/- 0.16). There was no death in the control and naloxone-treated groups. The incidences of death were 2/15 in F50 group, 5/15 in F100 group, and 7/15 in the F200 group (p < 0.05). The results of this study demonstrated that fentanyl potentiates the lidocaine-induced convulsions in a dose-dependent manner in mice, and this effect may be mediated by an opioid mechanism. PMID- 15897678 TI - Testosterone and nandrolone sensitization of brain anteroventral area of third ventricle to hypertonic NaCl-induced sympathetic response. AB - Androgenic steroids increase atherogenesis, thrombogenicity and endothelial dysfunction when administered in high doses, however their effects on NaCl sensitivity of the brain anteroventral area of the third ventricle (DeltaV3V) have not been explored. Sprague-Dawley male rats were anesthetized with sodium pentobarbital (40 mg/kg) and the femoral intra-arterial blood pressure and heart rate monitored through a strain-gauge blood pressure transducer and tachograph. DeltaV3V microinjections of (2 microl) 1.5 mol/l NaCl solution were done according to brain coordinates: AP = 7.0 mm, L = 1.0 mm, D = 7.5 mm through a 0.2 mm diameter stainless steel needle. The injection site was verified with 1.0 microl neutral red solution. Basal systolic blood pressure increased 37.6 and 39.6 mm Hg after testosterone (1 mg/kg/day for 20 days) and nandrolone (1 mg/kg/day for 20 days) treatment respectively; diastolic blood pressure also increased upon testosterone and nandrolone treatment in 36.4 and 53.1 mm Hg, respectively; basal heart rate did not change. Vasopressor response to 1.5 mol/l NaCl DeltaV3V microinjection was higher in testosterone-treated rats; systolic blood pressure increased 56.0 vs. 28.3 control mm Hg; diastolic blood pressure increased 54.0 vs. 25 control mm Hg. This hypertensive response was 29% longer lasting in testosterone compared to vehicle-treated rats. The same pattern of DeltaV3V sensitization to hypertonic NaCl was observed in nandrolone-treated rats. Blood lipid profile changed to a proatherogenic fashion upon testosterone and nandrolone long-term treatment; the plasma-free testosterone concentration increased from 4.9 +/- 0.9 to 36.0 +/-7.1 pg/ml with the same testosterone treatment schedule. In conclusion, long-term androgenic steroid treatment sensitizes the brain DeltaV3V region to hypertonic NaCl which in turn conducts into a sympathetic vasopressor and heart rate-stimulating action. PMID- 15897679 TI - Lysophosphatidic acid induces histamine release from mast cells and skin fragments. AB - The present study examined whether lysophosphatidic acid (LPA) induces histamine release and the role of Rho-associated protein kinase (ROCK) in histamine release utilizing Y-27632, an inhibitor of ROCK. Rat peritoneal mast cells and mouse skin fragments were challenged with LPA; subsequently, histamine contents were measured by spectrofluorometric assay. LPA-induced histamine release from mast cells and skin fragments occurred in a time- and dose-dependent manner Pretreatment with Y-27632 inhibited LPA-induced histamine release in a dose dependent fashion. LPA-induced histamine release was not influenced by calcium free conditions; however, A23187-induced histamine release decreased significantly. TMB-8, an intracellular calcium antagonist, dose-dependently inhibited the histamine release under these conditions. Additionally, Y-27632 scarcely affected A23187-induced histamine release. These findings suggest that LPA-induced histamine release may be attributable to calcium release from intracellular stores. Moreover, ROCK participates in the extracellular calcium independent process of LPA-induced histamine release. PMID- 15897680 TI - Alexithymic characteristics and patient-therapist interaction: a video analysis of facial affect display. AB - Alexithymia as a disorder of affect regulation entails a patient's reduced ability to process emotional information. The purpose of this study was to evaluate the impact of alexithymia [as measured by the Toronto Alexithymia Scale (TAS)-26, German version] on affective correlates in a dyadic therapeutic interaction (as recorded by the Emotional Facial Action Coding System). Interviews with 12 in-patients with various psychosomatic disorders (anxiety, depression, somatisation) were videotaped and evaluated for facial affect display. The corresponding emotional reactions of the therapists (split screen) were recorded separately. Patients with high alexithymia scores (TAS-26 total score) tended to display less aggressive affects than those with low scores. The therapists' predominant emotional reaction to alexithymic patients was contempt. Our findings underscore the deep-rooted nature of alexithymia as a disorder of affect regulation. Since facial affects play a major role in the regulation of emotional interaction, this disorder may evoke negative reactions of potential caregivers. PMID- 15897681 TI - Can teachers' behavior ratings be used to screen early adolescent boys for psychiatric diagnoses? AB - The present study examined if a short, 22-item teacher rating questionnaire could be used to screen young adolescent boys for psychiatric diagnoses. Subjects were 239 12-year-old boys from a community sample of low socioeconomic status families. Child and parent versions of the Diagnostic Interview Schedule for Children were used to provide DSM-III-R diagnoses. Results show a low to moderate screening value for the Short Social Behavior Questionnaire (S-SBQ) scales, with best results for externalizing disorders. The method used to establish the diagnoses had an impact on the screening efficacy of the S-SBQ, higher prevalence rates resulting in a lower proportion of false-positives for the same cutoff point. Additional research is needed to assess the screening efficacy of the S SBQ across ages and gender, but results suggest that teacher ratings with the S SBQ could be used to screen boys with externalizing disorders at the end of elementary school. PMID- 15897682 TI - Glucocorticoids for human skin: new aspects of the mechanism of action. AB - Topical glucocorticoids have always been considered first-line drugs for inflammatory diseases of the skin and bronchial system. Applied systemically, glucocorticoids are used for severe inflammatory and immunological diseases and the inhibition of transplant rejection. Owing to the progress in molecular pharmacology, the knowledge of the mechanism of action has increased during the last years. Besides distinct genomic targets, which are due to the activation of specific cytoplasmatic receptors resulting in the (trans-) activation or (trans-) repression of target genes, there are non-genomic effects on the basis of the interference with membrane-associated receptors as well as with membrane lipids. In fact, various glucocorticoids appear to differ with respect to the relative influence on these targets. Thus, the extended knowledge of glucocorticoid induced cellular signalling should allow the design and development of even more specifically acting drugs - as it has been obtained with other steroids, e.g. estrogens for osteoporosis prevention. PMID- 15897683 TI - Characterization of large-conductance Ca2+-dependent and -independent K+ channels in HaCaT keratinocytes. AB - To characterize ion channels expressed in cell membrane of human keratinocytes, patch-clamp recordings were carried out in HaCaT cells. Two types of large conductance K(+) channels (about 250 pS) were measured. One type was activated by micromolar concentrations of intracellular Ca(2+) ions ([Ca(2+)](i)) and membrane depolarization, the other was [Ca(2+)](i) independent. The channels were neither dependent on intracellular ATP nor Mg(2+) nor on membrane stretch. We conclude that HaCaT keratinocytes express Ca(2+)-dependent maxi K(+) channels and still unknown large Ca(2+)-independent K(+) channels. These K(+) channels may affect the proliferation and differentiation of human keratinocytes by the influence on the resting potential, which may control the Ca(2+) influx across the cell membrane. PMID- 15897684 TI - Antagonism of epidermal growth factor receptor tyrosine kinase ameliorates the psoriatic phenotype in organ-cultured skin. AB - Psoriatic plaque skin incubated for eight days in organ culture in the presence of a potent epidermal growth factor (EGF) receptor tyrosine kinase (RTK) antagonist reverted to a more normal histological appearance, while untreated psoriatic plaque skin retained histological features associated with the psoriatic phenotype. In concomitant studies it was shown that the EGF-RTK antagonist had no significant effect on histological features of non-psoriatic skin and no effect on dermal function, i.e. elaboration of both type I procollagen and matrix metalloproteinase-1 (MMP-1; interstitial collagenase). When human epidermal keratinocytes were treated with the EGF-RTK antagonist in monolayer culture, growth inhibition was seen (ED(50) = approximately 0.06 microM). When dermal fibroblasts were exposed to the EGF-RTK antagonist in monolayer culture, proliferation, MMP-1 and type I procollagen production were essentially unaffected at concentrations which interfered with keratinocyte growth (up to 1 microM). The capacity of the EGF-RTK antagonist to modulate the histological features of psoriatic skin in organ culture under conditions in which normal skin architecture and dermal function are largely unaffected suggests a potential for anti-psoriatic therapy. PMID- 15897685 TI - Investigations of interactions of chlormezanone racemate and its enantiomers on human keratinocytes and human leucoytes in vitro. AB - Chlormezanone is a centrally acting muscle relaxant introduced in human therapy as a racemic substance. The following investigation was performed in order to investigate whether the racemate and both enantiomers differ in their potential cytotoxicty in vitro. We investigated antiproliferative effects and cytotoxicity (PicoGreen and ATP assay) for human HaCaT keratinocytes, production of oxygen radicals (ROS) by human interleukin-3-stimulated leukocytes (Lucigenin assay) and production of sulfoleukotrienes (Cellular Antigen Stimulation Test - CAST) by human leukocytes. In the dosage range of 0.001 to 0.1 mg/ ml chlormezanone, no antiproliferative effects were measured with the racemate and both enantiomers. At 1.0 mg/ml, a decrease of proliferative activity was seen after 48 h incubation time of about 50% for the enantiomers and of about 80% for the racemate (PicoGreen) and 50% (enantiomers) or 21% (racemate) in the ATP assay, respectively. ROS production was significantly inhibited at concentrations < or =0.01 mg/ ml by the racemate and the (+)-enantiomer, whereas the (-)-enantiomer was less effective. There was no stimulation of sulfidoleukotrienes in human leukocytes by chlormezanone. Present data argue for absence of significant cytotoxicity against human HaCaT keratinocytes and a dose-dependent suppression of ROS production by human leukocytes that is not uniform among the racemate and its enantiomers. PMID- 15897686 TI - Transepidermal water loss dynamics of human vulvar and thigh skin. AB - Refinement in procedures to assess skin surface water loss (SSWL) dynamics of the vulvar skin on a large sample of subjects (60) is described and compared to another semi-occluded skin site, the inner thigh. Vulvar SSWL significantly decreased over a 30-min period from 46.2 +/- 2.6 (SE) to 24.7 +/- 1.6 g m(-2) h (p < 0.001). The inner thigh, another semi-occluded region, showed no similar pattern for SSWL (6.2 +/- 0.3 to 6.6 +/- 0.5 g m(-2) h), and the values were significantly less than those for vulvar skin. There was no significant effect of age, body mass index or atopic status on vulvar SSWL. PMID- 15897687 TI - Diclofenac-Na gel is effective in reducing the pain and inflammation associated with exposure to ultraviolet light - results of two clinical studies. AB - The potential of a diclofenac-Na Emulgel (diclofenac gel) to alleviate the pain and associated symptoms caused by sunburn has been evaluated versus vehicle. Sunburn was induced on the buttock skin of healthy adult male subjects by irradiation with UVA + UVB rays. Investigational products were applied 6 and 10 h after irradiation, and efficacy was assessed on the basis of spontaneous and provoked pain, erythema, oedema, skin colour and temperature. The minimal efficacious concentration evaluated in an extension (0.1 vs. 0.25% diclofenac gel) of a previous concentration-finding study (1, 0.5 and 0.25% diclofenac gel) was 0.1% and was efficacious in alleviating pain (spontaneous and provoked) as well as reducing erythema, oedema and skin temperature. In a single- versus 2 application comparison study, a single application of 0.1% gel was sufficient to alleviate the pain and accompanying symptoms of sunburn with an onset of action 2 h after application. A second application of gel 4 h after the first maintained the analgesia and reduction of other symptoms for a period of up to 48 h after irradiation. PMID- 15897688 TI - Coexistence of copy number changes of different genes (INK4A, erbB-1, erbB-2, CMYC, CCND1 and ZNF217) in urothelial tumors. AB - The aim of this study was to establish the frequency of combinatorial and separate copy number changes of INK4A (9p21), erbB-1 (7p11), erbB-2 (17q17-21), CMYC (8q24), CCND1 (11q13) and ZNF217 (20q13) in urothelial tumors; a tissue microarray of 159 urothelial bladder tumors was analyzed by fluorescence in situ hybridization. A total of 38 invasive tumors were successfully analyzed for all 6 loci. Normal gene copy numbers of all loci were established in 13 tumors (34.2%). In 25 tumors (65.8%), at least one aberration was found. Single abnormalities were detected in 16 tumors (64%), while double or higher abnormalities were found in 9 tumors (39%). The most frequent genetic change was deletion of INK4A (60% of aberrant tumors), followed by increased copy number changes of ZNF217 (36%), CCND1 (28%), CMYC (12%) and erbB-1 (4%). It was significantly more frequent in pT1 than in pT2-4 tumors and was predominantly found separately, while oncogene copy number increases were usually combined with another aberration and were not associated with the tumor stage. We concluded that INK4A loss is usually found as a single aberration in bladder cancer, which is more frequent in pT1 than in pT2 4 tumors. Overrepresentations of putative oncogenes are present in these two groups with similar frequency and are rarely found as single abnormality. PMID- 15897689 TI - Laparoscopic environmental changes during surgery enhance the invasive potential of tumours. AB - OBJECTIVE: The use of laparoscopic techniques in resection of malignant tumours has been proposed to offer potential benefit to the patient in the form of earlier recovery and less immune paresis; however, reported tumour seeding, both peritoneal and at port site, has put this approach into question. The biological effects of the introduction of carbon dioxide or helium to form a pneumoperitoneum on tumour invasion and dissemination are unknown. METHODS: A human colonic adenocarcinoma cell line (SW1222) was exposed to in vitro laparoscopic environment of either carbon dioxide or helium for 4 h, mimicking the duration of a laparoscopic colorectal resection. Alteration in production of matrix metalloproteinase (MMP)-2, MMP-9 and urokinase-type plasminogen activator (uPA) due to exposure to a laparoscopic environment was determined by zymography and correlated to invasive capacity by a standard Matrigel-based invasion assay. Incorporation of specific gelatinase inhibitors or antibodies directed at the uPA receptor was utilized to determine the relative importance of proteases. RESULTS: Exposure to the laparoscopic environment significantly enhanced production of the proteases MMP-2, MMP-9 and uPA. A concomitant enhancement of invasive capacity was also observed, being blocked by specific protease inhibitors. Changes in both protease production and aggression were observable for at least 24 h following the removal of the operative environment, indicating the possible long-term effects of the initial insult. CONCLUSION: Exposure to the laparoscopic environment enhances the invasive capacity of colonic adenocarcinomas via a well defined protease-determined pathway. It therefore appears likely that tumour cells released into the operative field can be made increasingly aggressive by a laparoscopic operative environment and can thus contribute to disease dissemination. PMID- 15897690 TI - Neural cell adhesion molecule-deficient beta-cell tumorigenesis results in diminished extracellular matrix molecule expression and tumour cell-matrix adhesion. AB - To understand by which mechanism neural cell adhesion molecule (N-CAM) limits beta tumour cell disaggregation and dissemination, we searched for potential downstream genes of N-CAM during beta tumour cell progression by gene expression profiling. Here, we show that N-CAM-deficient beta-cell tumorigenesis is associated with changes in the expression of genes involved in cell-matrix adhesion and cytoskeletal dynamics, biological processes known to affect the invasive and metastatic behaviour of tumour cells. The extracellular matrix (ECM) molecules emerged as the primary target, i.e. N-CAM deficiency resulted in down regulated mRNA expression of a broad range of ECM molecules. Consistent with this result, deficient deposition of major ECM stromal components, such as fibronectin, laminin 1 and collagen IV, was observed. Moreover, N-CAM-deficient tumour cells displayed defective matrix adhesion. These results offer a potential mechanism for tumour cell disaggregation during N-CAM-deficient beta tumour cell progression. Prospective consequences of these findings for the role of N-CAM in beta tumour cell dissemination are discussed. PMID- 15897691 TI - Can we prevent prostate cancer? Rationale and current status of prostate cancer chemoprevention. AB - Prostate cancer has been one of the most frequent cancers among men in Western countries for the past decade. Investigation of prostate cancer prevention is very attractive, because prostate cancer has a high incidence, long-term natural history, regional difference in incidence, and is effected by sex steroids. Chemoprevention is defined as the use of specific agents to suppress or reverse carcinogenesis and to prevent the development of cancer. The development of chemoprevention strategies against prostate cancer would be of medical and economic importance. Basic and clinical research of chemoprevention of prostate cancer are under active investigation. This article aims to summarize and review the basic evidence and clinical trials on prostate cancer chemoprevention. Recent research has demonstrated that many agents, such as agents altering sex steroid signaling, drugs inducing antiproliferation/differentiation, retinoids, anti inflammatory drugs, and antioxidants, could be potential preventatives for prostate cancer. Large-scale clinical trials have suggested that 5alpha-reductase inhibitor finasteride, selenium, and vitamin E can function as a chemopreventive agent. Although no definitely effective strategies of prostate cancer prevention have been identified yet, increasing evidence will provide effective and safe strategies that bring clinical benefits. PMID- 15897692 TI - Management of prolonged urinary leakage at the urethro-vesical anastomosis. AB - Urinary leakage at the urethrovesical anastomosis is a rare but troublesome complication after radical prostatectomy. Usually, the anastomosis is almost watertight when tested during the operation and small leakages typically seal off in a few days. We describe 3 cases with postoperative urinary leakage where we have utilized a simple procedure of replacing the standard Foley catheter with another catheter with extra side fenestrations close to the anastomosis. Side fenestrations provide efficient drainage at the anastomosis area leading to a rapid resolution of the leakage. The method described represents an effective and low-risk solution to a potentially harmful complication. PMID- 15897693 TI - Comparative effects of bicalutamide (Casodex) versus orchidectomy on bone mineral density, bone remodelling, and bone biomechanics in healthy rats. AB - INTRODUCTION: The aim of this work was to analyze the effects produced on bone mineral density (BMD) by the administration of bicalutamide and to compare them with those produced by orchidectomy. Bone formation rate (serum osteocalcin), bone resorption (serum carboxyterminal telopeptide of collagen I; CTX), and biomechanical properties of bone were also studied. METHODS: Thirty-eight male Wistar rats were used: (1) Sham group, rats sham operated at 16 weeks of age; (2) OQX group, rats orchidectomized at 16 weeks of age, and (3) Bic group, rats sham operated at 16 weeks of age and treated during 6 weeks with bicalutamide. The rats were sacrificed at 22 weeks of age, and the BMD in femur and lumbar spine was determined. Serum osteocalcin and serum CTX were also analyzed. Biomechanical parameters related to torsion assay were also studied. RESULTS: The OQX group showed a significant decrease in femoral BMD with respect to Sham rats, whereas bicalutamide treatment did not produce any significant change in BMD. Both Sham and Bic groups showed similar serum osteocalcin and CTX values, whereas OQX rats presented higher osteocalcin and CTX levels than the Sham group. The OQX group showed a significant decrease in femoral thickness. No significant differences were observed in the rest of the biomechanical parameters between groups. CONCLUSION: These results indicate that bicalutamide treatment, in spite of its anti-androgenic properties, does not affect bone remodelling nor BMD in male healthy rats, suggesting that this compound may function as a selective androgen receptor modulator for effects on bone remodelling in the osteoblasts. PMID- 15897694 TI - Efficacy of unilateral nerve sparing in radical perineal prostatectomy. AB - AIM: We determine the efficacy of unilateral nerve-sparing radical perineal prostatectomy in preserving the sexual function. PATIENTS AND METHODS: Ninety-two patients with histologically confirmed unilateral prostate cancer were scheduled for contralateral nerve preservation. The perioperative morbidity was assessed using the patients' chart reviews. Postoperative health-related quality of life, urinary continence, and potency were evaluated prospectively with questionnaires provided before surgery and then after 6, 12, and 24 months. RESULTS: Unilateral nerve preservation was performed in 88 of the 92 patients. Due to extensive scarring or prostatic size, the procedure was terminated as regular radical prostatectomy in 4 other patients. The perioperative complication rate was low and of minor significance, except in 1 patient who experienced a significant myoglobulinuria due to a prolonged procedure. Blood transfusions were necessary in 5 (5.4%) patients. Ureteral reimplantation was performed in 1 patient because of ureteral stricture. Positive surgical margins were present in 12 (18%) of 67 pT2 patients and in 8 (35%) of 23 pT3 patients. A proportion of 48% (15/31) of the patients followed for more than 24 months and who had a good erectile function prior to surgery reported unassisted sexual intercourse. However, only 4 of these patients were completely satisfied with all aspects of sexual performance, as asked in a short version of the International Index of Erectile Function questionnaire. CONCLUSIONS: Unilateral nerve-sparing radical perineal prostatectomy is technically feasible and yields excellent results in terms of potency preservation for prostates <60 ml. However, the quality of erections is decreased, even in patients with erections sufficient for intercourse. Hence, appropriate sexual counseling in conjunction with medical therapy should be offered to all patients. PMID- 15897695 TI - Association of vitamin D receptor gene polymorphism and risk of prostate cancer in India. AB - INTRODUCTION: Vitamin D plays an important role in the proliferation and differentiation of normal and malignant cells. In several studies polymorphism in the vitamin D receptor (VDR) gene has been reported to be associated with prostate cancer (CaP). The rationale of this study was to determine the association between the VDR (Fok-I) polymorphism and the risk of developing CaP. MATERIALS AND METHODS: Polymorphism was detected by the polymerase chain reaction (PCR)-restriction fragment length polymorphism method in 128 CaP patients (age range 43-89 years) and 147 age-matched controls (age range 42-91 years). PCR products were designated as F or f allele according to the absence or presence of a restriction site. RESULTS AND CONCLUSIONS: The frequencies of the FF, Ff and ff genotypes were 60.9, 35.2 and 3.9% in CaP patients and 42.2, 46.9 and 10.9% in healthy controls, respectively. The genotype frequency distribution between CaP and the control group was statistically significant (p = 0.003). However, the distribution of genotypes was not significantly associated with the Gleason score. The present study thus demonstrates that the FF genotype (or F allele) of the VDR gene plays an important role in determining the risk of CaP and could be postulated as a good candidate genetic marker. PMID- 15897696 TI - Ureteroscopic management of sepsis associated with ureteral stone impaction: is it still contraindicated? AB - INTRODUCTION: Retrograde decompression is generally not advocated for patients with sepsis owing to ureteral obstruction by stone impaction, and the initial treatment of choice is percutaneous nephrostomy (PCN). We report our experience with the treatment of urosepsis with retrograde ureteroscopy (URS) instead of PCN drainage. PATIENTS AND METHODS: Fifty-six consecutive patients diagnosed with ureteral stone-related sepsis received URS as primary treatment at our institution. Patients with uncontrollable sepsis underwent emergent URS and hemodynamically stable patients underwent elective URS within two days of diagnosis. RESULTS: URS was successful in 53 (94.6%) of the 56 patients. PCN was performed in the 3 cases of URS failure. Internal ureteral stenting was performed in 48 patients. Secondary procedures were performed in 10 (18.9%) patients. Twenty-six patients suffered from postoperative fever for an average of 1.6 days (range 1-4 days). There were no anesthesia-related morbidities, postoperative exacerbations of the clinical condition, or postoperative deaths. The median length of hospital stay was 7 days (range 3-94 days). CONCLUSION: PCN drainage is the standard treatment of sepsis associated with ureteral stone obstruction. However, our results show that URS can be safely and successfully performed by skilled endourologists in select clinical situations. PMID- 15897697 TI - Risk factors for the formation of a steinstrasse after shock wave lithotripsy. AB - INTRODUCTION: We studied the various stone, renal, and therapy factors that could affect steinstrasse formation after shock wave lithotripsy (SWL) to define their predictive value. PATIENTS AND METHODS: Between May 1999 and September 2002, 563 patients were treated with a Stonelight V3 lithotriptor. A steinstrasse was recorded in 46 patients. All patient data, stone and renal characteristics, and data of SWL were reviewed. Statistical analyses of patients, stones, and therapy characteristics in correlation with the incidence of steinstrasse formation were performed to assign factors that had a significant impact on the formation of this complication. RESULTS: The overall incidence of a steinstrasse was 8.17%. The steinstrasse was in the pelvic ureter in 84.3% of the cases, in the iliac ureter in 7.84% of them, and in pelvic and iliac ureter in 7.84% of the patients. The incidence of a steinstrasse significantly correlated with stone size and site. The incidence rates of a steinstrasse in renal stones <1 cm, 1-2 cm, and >2 cm were 4.46, 15.87, and 24.3% respectively. The incidence rates of this complication in ureteral stones <1 cm and 1-2 cm were 3.37 and 9.52%, respectively. The incidence rates of a steinstrasse in stones located in upper calices, middle calices, lower calices, and renal pelvis were 6.12, 10.52, 6.36, and 19.32%, respectively. CONCLUSIONS: Stone size and site are the significant factors predicting the formation of a steinstrasse. If a patient has a high probability of steinstrasse formation, close follow-up with early intervention or prophylactic pre-SWL ureteral stenting is indicated. PMID- 15897698 TI - Prulifloxacin versus ciprofloxacin in the treatment of adults with complicated urinary tract infections. AB - INTRODUCTION: The present study was performed to evaluate the efficacy and safety of a 10-day regimen of prulifloxacin 600 mg once daily as compared to ciprofloxacin 500 mg twice daily in the treatment of patients with complicated urinary tract infections (UTIs). MATERIALS AND METHODS: 257 patients (mean age +/ SD 62.3 +/- 16.5) were enrolled and orally treated with prulifloxacin (127 patients) or ciprofloxacin (130 patients). The study was designed as a randomized, double-blind, double-dummy, controlled clinical trial. The primary efficacy parameter was the eradication of infecting strains (<10(3) cfu/ml). The clinical outcome and tolerability were also assessed. RESULTS: At baseline, the most common infecting strains were Escherichia coli (62.8%), Proteus mirabilis (7.1%) and Klebsiella pneumoniae (4.1%). At the early follow-up, the rate of patients showing successful treatment was 90.8% in the prulifloxacin group, and 77.8% in the ciprofloxacin group (p = 0.008). A positive clinical outcome was observed in 94.8 and 93.3% of prulifloxacin- and ciprofloxacin-treated patients. Both drugs were well tolerated. Two patients dropped out for treatment-related adverse events. CONCLUSIONS: The high urinary concentrations of prulifloxacin, combined with a broad-spectrum antimicrobial activity, allow its use in the empiric therapy of UTIs. PMID- 15897699 TI - Should high-grade prostatic intraepithelial neoplasia change our approach to infravesical obstruction? AB - OBJECTIVES: To investigate whether coexistence of high-grade prostatic intraepithelial neoplasia (HPIN) should change our therapeutic approach to infravesical obstruction. MATERIAL AND METHODS: Of 505 patients who underwent sextant transrectal ultrasonography (TRUS)-guided prostate biopsy, 65 (12.8%) had HPIN and 29 of them underwent prostatectomy (23 transurethral resection of prostate (TURP), 6 open) due to obstructive urinary symptoms. Patients without carcinoma were followed up with semiannual prostate-specific antigen (PSA) and digital rectal examination. After a follow-up of 24.8 +/- 11.0 months, 19 of 29 patients who accepted our call had another sextant biopsy. RESULTS: Mean age and initial mean PSA values of 29 patients were 67.6 +/- 6.7 years and 9.26 +/- 5.91 ng/ml, respectively. The final pathological evaluation of the surgical specimens revealed 2 prostatic adenocarcinomas both in the TURP group. The remaining 27 (93.2%) patients were found to have benign prostatic hyperplasia (BPH) and their serum PSA levels declined from 9.26 +/- 5.91 to 4.59 +/- 2.0 ng/ml 3 months after prostatectomy. Of the 19 patients who had another biopsy with a mean PSA value of 4.06 +/- 4.61 ng/ml, 15 and 4 of them had BPH and HPIN respectively. CONCLUSIONS: Our preliminary data indicate that the presence of HPIN on TRUS-guided biopsies is not a factor to delay an indicated surgical intervention for infravesical obstruction. PMID- 15897700 TI - Does ischemia-induced prostate damage during cardiac surgery involving cardiopulmonary bypass cause bladder outlet obstruction? AB - OBJECTIVE: This study sought to investigate whether ischemia-induced prostate damage during cardiac surgery involving cardiopulmonary bypass causes bladder outlet obstruction. MATERIALS AND METHODS: The study involved 37 men who underwent elective cardiac surgery involving cardiopulmonary bypass. Prostate specific antigen (PSA) levels were determined preoperatively (baseline) and on postoperative days 1, 5, and 30. In 4 cases, the PSA level after the operation was unchanged from the preoperative level, so these 4 men were excluded from the study. In the remaining 33 patients, symptoms of bladder outlet obstruction were assessed using the International Prostate Symptom Score. Each subject completed this test preoperatively and 3, 6 and 9 months postoperatively, and the means scores at these time points were compared. The effects of patient age, operative time, CPB time, and aortic clamping time on postoperative increases in PSA levels were investigated. RESULTS: Thirty-three (89.2%) of the 37 men exhibited increased postoperative PSA levels compared to baseline. The mean PSA level for the 33 cases on day 5 was significantly higher than the baseline mean, but the mean levels on postoperative days 1 and 30 were comparable to baseline. Nine (24.3%) of the 33 men had postoperative PSA levels greater than 4.0 ng/dl (the upper normal limit). There was no significant difference between preoperative and postoperative International Prostate Symptom Scores. CONCLUSION: The study indicates that men's PSA levels are, indeed, increased after cardiac surgery with cardiopulmonary bypass. However, in 9 months of follow-up, there was no association between this PSA rise and development of BOO, according to International Prostate Symptom Scores. PMID- 15897701 TI - Is bladder cycling useful in the urodynamic evaluation previous to renal transplantation? AB - OBJECTIVES: To evaluate the usefulness of bladder cycling (BC) in differentiating between urodynamic abnormalities due to reversible bladder dysfunction and those due to preexisting lower urinary tract (LUT) abnormalities, and to determine if BC increases the diagnostic yield of urodynamic investigations (UIs) in these patients. METHODS: Eleven patients with oligoanuria (<300 ml/day) were evaluated with complete UIs. All cases showed low compliance (mean 6 ml/cm H2O) and low cystometric capacity (mean 146 ml) and were placed in a BC program. The indications for UIs were LUT symptoms in 2 patients, LUT abnormalities (myelodysplasia or urethral valves) in 3 patients, or the appearance of an extremely small bladder on cystogram (6 patients). BC was performed through a hypogastric catheter in an outpatient setting. UIs were repeated afterwards. RESULTS: After BC, 5 patients (45%) continued to have low compliance (<8 ml/cm H2O) and 4 patients (36%) had normal UIs. In 2 cases (18%), a previously unrecognized obstruction was diagnosed. No patient with LUT symptoms or abnormalities had a normal UI after BC. Conversely, 4 of 6 patients in whom BC was indicated due to a small bladder on cystogram had normal UIs after BC. CONCLUSIONS: BC is useful in explaining whether high bladder pressure is due to dysfunction or to preexisting disorders, and thus indicating the proper surgery for correction. Suprapubic BC significantly increases bladder capacity thus increasing the diagnostic yield of the UI. PMID- 15897702 TI - Tinea cruris in routine urology practice. AB - INTRODUCTION: Tinea cruris is almost exclusively a male dermatophytosis. This infection is also sexually transmitted, and may cause epidemics in public areas such as common bathing facilities, dormitories and among military recruits. It has been aimed to investigate the prevalence and the causative agents of tinea cruris and tinea pedis in patients who were admitted to the Department of Urology with various pathologies. MATERIALS AND METHODS: Direct microscopy and cultures of the epithelial scrapings were performed to identify the causative agent in patients who were admitted to the Department of Urology. RESULTS: Out of a total of 155 cases examined, 39 (25.1%) were mycologically proven cases of dermatomycosis. In 11 (28.2%) of the patients tinea pedis, in 3 (7.7%) tinea cruris and in 10 (25.6%) Candida intertrigo were detected. In the remaining 15 (38.5%) cases, only direct microscopic examinations were found positive. The most common causative agent was Candida albicans (33.4%) followed by Trichophyton rubrum (29.1%), Trichophyton mentagrophytes var. interdigitale (29.1%), Candida glabrata (4.2%) and Candida tropicalis (4.2%). CONCLUSION: The investigation of genital dermatomycosis should be a part of routine urological examination and the clinical diagnosis should be confirmed by mycological methods. PMID- 15897703 TI - Apoptosis in non-tumorous adult human testis tissue. Comparison of so-called 'normal' testis tissues. AB - INTRODUCTION: Apoptosis seems to play an important role in tumorigenesis, prognosis and therapy of testicular tumors. To understand its biological significance, it is important to quantify the amount of apoptosis and to compare the rate of apoptosis to that of a normal, unaffected reference tissue. Usually tissue from the unaffected site of the testis in patients with testicular cancer or testis tissue from patients who underwent surgical castration due to prostate cancer is used as the reference tissue. However it is not known, if both tissues are equivocal with respect to their apoptotic index. The purpose of the study was to compare the two most often used reference tissues for the quantification of apoptosis in testicular tissues with regard to their apoptotic index. MATERIALS AND METHODS: The apoptotic indices of both tissues were compared, using two standard apoptosis detection methods, i.e. in situ end labeling and a morphological approach. RESULTS: The apoptotic index in testis tissue from patients who were surgically castrated for anti-hormonal treatment of prostate cancer was shown to be significantly higher than the apoptotic index of tumor free but tumor-associated testicular tissue of testis cancer patients. There was a strong relationship between the apoptotic index and the age of the patients. CONCLUSION: Although there might be genetic changes in the tumor-associated testicular tissue influencing the apoptotic index, it seems advisable to use tumor-associated tissue rather than testis tissue of patients with prostate cancer as the reference tissue, due to the significant age dependence of the apoptotic index. PMID- 15897704 TI - Possibilities and limitations of fibrin glue usage in nephron-sparing surgery: experimental study. AB - INTRODUCTION: The possibilities and limitations of fibrin glue (FG) usage in nephron-sparing surgery were studied. MATERIALS AND METHODS: A prospective experimental study was carried out in 50 pigs: 30 with polar resection, and 20 with mediorenal wedge resection of the kidney. Hemostatic sutures, FG, and FG with a muscle 'cup' in animals with polar resection of the kidney were compared. FG and sutures in animals with the wedge resection of the kidney were studied as well. Bleeding, hot ischemia time, complication rate, and additional scarring were also analyzed. RESULTS: Suture hemostasis is safe but with significant adverse effects in both polar and wedge resection of kidney. FG was not efficient as a sole hemostatic agent for polar resection. It was as efficient as hemostatic suture for wedge resection of the kidney. FG with a muscle 'cup' on a pole of the kidney achieved good results in animals with polar resection of the kidney. Histological analysis confirmed better results with FG because of both the less intense and smaller area of additional scarring. CONCLUSION: FG is a reliable and efficient hemostatic agent for nephron-sparing surgery whenever both sided gluing is possible. PMID- 15897705 TI - Intravesical heparin and peripheral neuromodulation on interstitial cystitis. AB - INTRODUCTION: We wanted to evaluate the therapeutic effect of intravesical heparin and peripheral neuromodulation on patients with interstitial cystitis. MATERIALS AND METHODS: From March 2002 to August 2003, 8 female and 2 male subjects conform to the NIDDK criteria and not responsive to the previous conventional treatments were included in the study. Wisconsin pain scores, maximal cystometric capacities, and night and day voiding frequencies were determined and these studies were repeated in the 2nd and 12th months of the treatment with 10,000 units intravesical heparin and peripheral neuromodulation. Frequency of the treatment was once a week during first 8 weeks, once in 2 weeks in the following 8 weeks, and once in 3 weeks four times. Then, it was decreased to once a month. RESULTS: The mean follow-up period was 13 months (12-16 months). Day and night voiding frequency were significantly better in the 2nd and 12th months, when compared to pretreatment values. The Wisconsin pain scores were 62.5 +/- 13.9% and 62.8 +/- 15.2% in the 2nd and the 12th months, respectively. The average increase in the maximum cystometric capacity was 54.8 +/- 27.4% and 52.5 +/- 31.6% in the 2nd and the 12th months, respectively. CONCLUSIONS: Intravesical heparin and peripheral neuromodulation combination seems to be an alternative for patients with interstitial cystitis not responsive to other treatments. PMID- 15897706 TI - Pseudoexstrophy with epispadias. AB - The exstrophy variants are uncommon anomalies. The variants of the exstrophy complex have all the stigmata of the classical exstrophy such as divergent recti, widened symphysis pubis, and low-set umbilicus; however, the urinary tract is intact to a varying degree. Pseudoexstrophy, an exstrophy variant, is very rarely associated with epispadias. We report an unusual case of pseudoexstrophy with epispadias, in whom the intact bladder was initially covered by a mucous membrane which later epithelialized. The epispadias was repaired using a penile disassembly technique with posterior and ventral placement of bladder neck and urethra. Although the urethral meatus was hypospadiac, the child had achieved dry intervals with occasional stress incontinence. PMID- 15897707 TI - Gastric cancer in augmentation gastrocystoplasty. AB - A 27-year-old man operated for myelomeningocele in the neonatal period and for augmentation gastrocystoplasty at age 13 years for voiding dysfunction, was diagnosed with gastric adenocarcinoma on the stomach side of bladder augmentation. A radical cystectomy and an ileal conduit procedure were performed and the pathologist confirmed a poorly differentiated gastric adenocarcinoma. Urologists should be aware of this potential degeneration, and long-term follow up is mandatory to detect malignization of these cystoplasties. PMID- 15897708 TI - Endourologic management of duodenal calculi in pancreas-kidney transplantation. AB - Duodenal stones formed during pancreas-kidney transplantation are usually associated with nonabsorbable sutures or staples. We report on the delayed formation of a struvite duodenal stone not attributed to foreign material, managed successfully with intracorporeal electrohydraulic lithotripsy. PMID- 15897709 TI - Severe hypospadias associated with Robertsonian translocation. AB - In this study, we report a 3-year-old boy with severe scrotal hypospadias with Robertsonian translocation [45,XY,t(13q;14q)]. The patient was born at term with a low birth weight and hypospadias. There was no endocrinological abnormality. His father also has a balanced 13-14 Robertsonian translocation. Two-stage hypospadias repair was carried out. The presence of this chromosomal anomaly and hypospadias are unique to our patient, compared to others with the 45,XY,t(13q;14q) translocation. Although no such association has been reported so far, we thought that severe hypospadias in this case might be associated with this translocation. PMID- 15897710 TI - Sudden unexpected death in epilepsy: is death by seizures a cardiac disease? AB - RESULTS: Sudden unexpected death in epilepsy (SUDEP) gains more and more acknowledgment across the various interdisciplinary fields. Accordingly, we performed in a prospective setting a case-control study of all SUDEP cases in a well-defined part of Denmark (Northern Jutland), between January 1998 and September 2000. We attempted to look into the cardiopathologic mechanism behind this phenomenon by assessing the degree of myocardial fibrosis in SUDEP patients versus controls. The histologic evaluation was possible in 65% of the cases (15/23) whose death was attributed to SUDEP and in 71% (15/21) of controls. Forty percent of the SUDEP cases (6/15) presented several foci of fibrotic changes in the deep and subendocardial myocardium in contrast to 1 control (6.6%, P = 0.03). None of the subjects from the SUDEP group showed fibrotic changes in their conduction system as compared with 1 control (6.6%). The quantitative evaluation of fibrosis demonstrated a trend toward more fibrosis in the deep and subendocardial myocardium of the SUDEP cases. Forty percent of cases in the SUDEP group were men (6/15), characteristically young at time of death (mean age 38 years) and with a late epilepsy onset (mean age 21 years). Antemortem, 73% of the SUDEP patients (11/15) had experienced infrequent seizures (self-reported). We conclude that the SUDEP cases displayed significant fibrosis of the myocardium when this was assessed by qualitative means. This fibrosis may be the consequence of myocardial ischemia as a direct result of repetitive epileptic seizures, which, associated with the ictal sympathetic storm, may lead to lethal arrhythmias. PMID- 15897712 TI - Alcohol and mitochondria in cardiac apoptosis: mechanisms and visualization. AB - Apoptosis of myocytes is likely to contribute to a variety of heart conditions and could also be important in the development of alcoholic heart disease. A fundamental pathway to apoptosis is through mitochondrial membrane permeabilization and release of proapoptotic factors from the mitochondrial intermembrane space to the cytosol. The authors' results show that prolonged exposure of cultured cardiac cells to ethanol (35 mM for 48 hr) promotes Ca2+ induced activation of the mitochondrial permeability transition pore (PTP). PTP dependent mitochondrial membrane permeabilization is followed by release of cytochrome c and execution of apoptosis. The authors propose that chronic ethanol exposure, in combination with other stress signals, may allow for activation of the PTP by physiological calcium oscillations, providing a trigger for cardiac apoptosis during chronic alcohol abuse. Coincidence of apoptosis promoting factors occurs in only a small fraction of myocytes, but because of the absence of regeneration, even a modest increase in the rate of cell death may contribute to a decrease in cardiac contractility. Detection of apoptotic changes that are present in only a few myocytes at a certain time in the heart is not feasible with most of the apoptotic assays. Fluorescence imaging is a powerful technology to visualize changes that are confined to a minor fraction of cells in a tissue, and the use of multiphoton excitation permits imaging in situ deep in the wall of the intact heart. This article discusses potential mechanisms of the effect of alcohol on mitochondrial membrane permeabilization and visualization of mitochondria-dependent apoptosis in cardiac muscle. PMID- 15897713 TI - A novel single nucleotide polymorphism of the neuropeptide Y (NPY) gene associated with alcohol dependence. AB - BACKGROUND: Neuropeptide Y (NPY) is a major endogenous regulator of anxiety related behaviors and emotionality. Transgenic work with NPY and null-mutant mice have implicated NPY in the control of alcohol consumption, suggesting that genetic variation of the prepro-NPY gene may also contribute to the heritability of alcoholism. The aim of this study was to examine whether polymorphic variants of the NPY gene are associated with the diagnosis of alcohol dependence. METHODS: We compared allele frequencies of 5 NPY polymorphisms (-883-ins/del, -602, -399, 84, and +1128) in a Nordic population of alcohol-dependent individuals (n = 428 males; n = 149 females) and ethnically matched controls (n = 84 males; n = 93 females) for whom alcohol dependence or any diagnosis of substance disorder was excluded. Patients were further subtyped into type I (late-onset) and type II (early-onset) alcoholics. RESULTS: The -602 marker showed a significant association with alcohol dependence (p = 0.0035; OR, 2.3; 95% CI, 1.3-4.0); a trend level association was further observed for the -399 marker (p = 0.058; OR, 1.3; 95% CI, 0.99-1.7) and the +1128 marker (p = 0.053; OR, 1.8; 95% CI, 0.99 3.1). The association for the -602 marker remained and was strengthened when analyzed in type I subjects only, although this association was not seen in type II patients, and there also was a significant association in the female subjects but not in males. The -602 single nucleotide polymorphism was in strong linkage dysequilibrium (r2 = 0.7; p < 0.0001) with the +1128 single nucleotide polymorphism, which has previously been reported to be associated with a diagnosis of alcoholism. Haplotype-based association confirmed these results. CONCLUSIONS: We report a novel polymorphism at position -602 in the 5' region of the NPY gene that is significantly associated with alcohol dependence. We also describe the haplotype frequencies and linkage dysequilibrium pattern of four variations in that region. PMID- 15897714 TI - The syntaxin binding protein 1 gene (Stxbp1) is a candidate for an ethanol preference drinking locus on mouse chromosome 2. AB - BACKGROUND: We previously mapped a quantitative trait locus (QTL) for ethanol preference drinking to mouse chromosome 2 (mapped with high confidence, LOD = 15.5, p = 3 x 10(-16)). The specific gene(s) in the QTL interval responsible for phenotypic variation in ethanol preference drinking has not been identified. METHODS: In the current study, we investigated the association of the syntaxin binding protein 1 gene (Stxbp1) with ethanol preference drinking and other ethanol traits using a panel of B6 x D2 (BXD) recombinant inbred (RI) strains derived from the C57BL/6J (B6) and DBA/2J (D2) inbred mouse strains. Confirmation analyses for ethanol consumption and withdrawal were performed using a large B6D2 F2 cross, short-term selected lines derived from the B6 and D2 progenitor strains, and standard inbred strains. RESULTS: BXD RI strain analysis detected provisional associations between Stxbp1 molecular variants and ethanol consumption, as well as severity of acute ethanol withdrawal, ethanol-conditioned taste aversion, and ethanol-induced hypothermia. Confirmation analyses using three independent genetic models supported the involvement of Stxbp1 in ethanol preference drinking but not in ethanol withdrawal. CONCLUSIONS: Stxbp1 encodes a Sec1/Munc18-type protein essential for vesicular neurotransmitter release. The present study provides supporting evidence for the involvement of Stxbp1 in ethanol preference drinking. PMID- 15897715 TI - Comparison of basal neuropeptide Y and corticotropin releasing factor levels between the high ethanol drinking C57BL/6J and low ethanol drinking DBA/2J inbred mouse strains. AB - BACKGROUND: Recent genetic and pharmacological evidence indicates that low neuropeptide Y (NPY) levels in brain regions involved with neurobiological responses to ethanol promote increased ethanol consumption. Because of their opposing actions, it has been suggested that NPY and corticotropin releasing factor (CRF) exert a reciprocal regulation on drug self-administration. It has been widely reported that inbred C57BL/6 mice consume significantly higher amounts of ethanol than do DBA/2 mice. Therefore, we used immunohistochemical techniques to determine if basal NPY and/or CRF levels differed in predicted directions between C57BL/6J and DBA/2J mice. METHODS: Ethanol-naive C57BL/6J and DBA/2J mice were deeply anesthetized with sodium pentobarbital (100 mg/kg) and perfused transcardially with 0.1 mM of phosphate-buffered saline followed by 4% paraformaldehyde in buffered saline. Brains were collected and postfixed for 4 hr at 4 degrees C and then were cut into 35-microm sections. Tissues containing the nucleus accumbens (NAc), hypothalamus, and amygdala were processed for NPY or CRF immunoreactivity using immunofluorescent or DAB techniques. Immunoreactivity was quantified from digital images using Image J software. RESULTS: The C57BL/6J mice showed reduced NPY expression in the NAc shell, the basolateral amygdala, and the central nucleus of the amygdala when compared with DBA/2J mice. However, these strains did not differ in CRF expression in any of the brain regions analyzed. CONCLUSIONS: These data suggest that low NPY levels in the amygdala and/or the shell of the NAc, which are not compensated for by similar changes in CRF levels, may contribute to the high ethanol consumption characteristic of C57BL/6J mice. PMID- 15897716 TI - Decreased oral self-administration of alcohol in kappa-opioid receptor knock-out mice. AB - BACKGROUND: Although a large body of evidence suggests a role for the opioid system in alcoholism, the precise role of mu-, delta-, kappa-, and ORL1-opioid receptors and the physiological significance of their natural genetic variation have not been identified. The method of targeted gene disruption by homologous recombination has been used to knock out (KO) genes coding for opioid receptors, and study their effects on alcohol self-administration. Here we examined the effects of targeted disruption of kappa-opioid receptor (KOR) on oral alcohol self-administration and other behaviors. METHODS: Oral alcohol, saccharin and quinine self-administration was assessed in a two-bottle choice paradigm using escalating concentrations of alcohol, or tastant solutions. In preference tests 12% alcohol, 0.033% and 0.066% saccharin, and 0.03 mM and 0.1 mM quinine solutions were used. Open-field activity was determined in an arena equipped with a computer-controlled activity-detection system. Subjects were tested for three consecutive days. Locomotor activity was assessed on days 1 and 2 (after saline injection, i.p.) and on day 3 (after alcohol injection, i.p.). Alcohol-induced locomotor activity was determined as the difference in activity between day 3 and day 2. RESULTS: Male KOR KO mice in preference tests with 12% alcohol consumed about half as much alcohol as wild-type (WT) or heterozygous (HET) mice, showed lower preference for saccharin (0.033% and 0.066%) and higher preference to quinine (0.1 mM) than WT mice. Female KOR KO mice showed similar reduction in alcohol consumption in comparison to WT and HET mice. Partial deletion of KOR in HET mice did not change alcohol consumption in comparison to WT mice. In all genotype-groups females drank significantly more alcohol than males. MANOVA of locomotor activity among KO, WT, and HET mice indicated that strain and sex effects were not significant for alcohol-induced activation (p > 0.05), while strain x sex interaction effects on alcohol-induced activation could be detected (F(1,55) = 6.07, p < 0.05). CONCLUSION: Our results indicating decreased alcohol consumption, lower saccharin preference, and higher quinine preference in KOR KO mice are in line with previous observations of opioid involvement in maintenance of food intake and raise the possibility that the deficient dynorphin/KOR system affects orosensory reward through central mechanisms which reduce alcohol intake and disrupt tastant responses, either as direct effects of absence of kappa opioid receptors, or as effects of indirect developmental compensatory changes. PMID- 15897717 TI - Kinetics of homocysteine metabolism after moderate alcohol consumption. AB - BACKGROUND: Moderate alcohol consumption is associated with a decreased risk of cardiovascular disease. Because plasma homocysteine (tHcy) is considered an independent risk factor for cardiovascular disease and associated with alcohol consumption, the authors investigated the effect of moderate alcohol consumption on kinetics of plasma tHcy concentration, vitamin B status, and other parameters involved in tHcy metabolism. METHODS: Ten healthy men and nine healthy postmenopausal women (aged 45-65 years) participated in a randomized, diet controlled, crossover trial. They consumed beer or alcohol-free beer (men: 4 units/day; women: 3 units/day) during 3 weeks, separated by a 1-week washout. On days 5, 10, 15, and 20 of each period, fasting blood samples were taken. RESULTS: Plasma tHcy (microM) and S-adenosyl methionine/S-adenosyl homocysteine ratio were not affected by consumption of beer or alcohol-free beer (p = 0.33 and p = 0.14, respectively). Plasma pyridoxal-5-phosphate (microg/liter) increased during consumption of beer (+11.0%), whereas it decreased during consumption of alcohol free beer (-34.0%; p = 0.042). Changes over time of plasma vitamin B6 (microg/liter) were similar to changes in plasma pyridoxal-5-phosphate (p = 0.10). Serum vitamin B12 was higher (p < 0.001) after 3 weeks consumption of alcohol-free beer (382.8 +/- 23.7 pg/liter) as compared with beer consumption (327.5 +/- 22.2 pg/liter). Changes in serum methionine, cysteine, cystathionine, and plasma folate were not different between beer-drinking and alcohol-free beer drinking periods. CONCLUSIONS: This study shows that moderate alcohol consumption does not affect plasma tHcy concentrations or S-adenosyl methionine/S-adenosyl homocysteine ratio. However, it does increase plasma vitamin B6 and decrease serum vitamin B12. PMID- 15897718 TI - Acute ethanol decreases dopamine transporter velocity in rat striatum: in vivo and in vitro electrochemical measurements. AB - BACKGROUND: Ethanol increases dopamine transporter (DAT) velocity when measured in cell expression systems, but its effects in vivo are mixed. The present experiments examined the effect of acute ethanol on dopamine transmission, particularly DAT velocity, in anesthetized animals as well as rat striatal suspensions. METHODS: To determine the effect of acute ethanol on DAT function in vivo, we measured dopamine uptake in real time using fast-scan cyclic voltammetry and constant potential amperometry in the olfactory tubercle of anesthetized rats. Dopamine fibers were electrically stimulated, and the resulting transient dopamine signals were analyzed to describe the release and uptake kinetics. We also measured the effect of ethanol on DAT velocity in vitro in striatal tissue suspensions using rotating disk electrode voltammetry. RESULTS: Ethanol (2.5 and 4 g/kg, intraperitoneally) decreased the electrically stimulated dopamine signal in the olfactory tubercle by 35-55%. The slope of the clearance curve of dopamine was 40% shallower after both doses of ethanol, indicating slower uptake. Modeling the data using Michaelis-Menten uptake kinetics showed that the slower uptake was due to a decrease in DAT V(max). These results were confirmed in vitro, because ethanol decreased the velocity of dopamine uptake by 35% in striatal tissue suspensions. CONCLUSIONS: These results indicate that acute ethanol decreases DAT function in rat dorsal and ventral striatum in anesthetized rats and tissue suspensions, in contrast to its effects on human DAT expressed in single cells. Given the variety of molecular targets of ethanol in the brain, including the DAT itself, it is likely that several mechanisms converge to produce a net effect on DAT regulation and function that could very well be different in intact tissue versus single cells. PMID- 15897719 TI - An extract of the Chinese herbal root kudzu reduces alcohol drinking by heavy drinkers in a naturalistic setting. AB - BACKGROUND: Of the available medications for treating alcohol-related problems, none are universally effective, and all have side effects that may limit their use. Extracts of kudzu containing a variety of isoflavones have been shown to reduce alcohol drinking in rats and hamsters. METHODS: The present study was designed to test the efficacy of a kudzu extract in a clinical population. Male and female "heavy" alcohol drinkers were treated with either placebo or a kudzu extract for 7 days and then given an opportunity to drink their preferred brand of beer while in a naturalistic laboratory setting. Participants served as their own controls, and order of treatment exposure was counterbalanced. Drinking behavior was monitored by a digital scale that was located in the top of an end table. RESULTS: Kudzu treatment resulted in significant reduction in the number of beers consumed that was paralleled by an increase in the number of sips and the time to consume each beer and a decrease in the volume of each sip. These changes occurred in the absence of a significant effect on the urge to drink alcohol. There were no reported side effects of kudzu treatment. CONCLUSION: These data suggest that an extract of this leguminous plant may be a useful adjunct in reducing alcohol intake in a naturalistic setting. PMID- 15897720 TI - Alpha-synuclein protein levels are increased in alcoholic patients and are linked to craving. AB - BACKGROUND: Alpha synuclein has been found to be increased in dopamine neurones of cocaine abusers and in rats whose alcohol preference is inbred. Furthermore, increased alpha-synuclein messenger RNA expression has been linked to craving in patients with alcoholism. The aim of the current study was to investigate whether protein levels of alpha synuclein in alcoholics are changed and possibly influence alcohol craving. METHODS: The alpha-synuclein protein expression level was measured by enzyme-linked immunosorbent assay in the serum of 49 male alcoholics and 50 nondrinking healthy controls. Alcohol craving was assessed by the Obsessive-Compulsive Drinking Scale total score, including subscales for obsessive and compulsive craving. RESULTS: Alpha-synuclein protein expression in patients with alcoholism (14.33 ng/ml; SD, 13.01 ng/ml) was significantly higher (t test, T = 3.66, p < 0.0001) when compared with that of healthy controls (5.92 ng/ml; SD, 9.72 ng/ml). Using a multivariate analysis, all craving scores (Obsessive-Compulsive Drinking Scale total score and obsessive and compulsive subscale scores) in alcoholics were significantly associated with their alpha synuclein protein levels (multiple linear regression, p < 0.014). CONCLUSIONS: To our knowledge, this is the first study evaluating alpha-synuclein protein expression in alcoholics. The current study provides further evidence of altered alpha-synuclein levels in patients with alcoholism and their linkage to alcohol craving. Because alpha synuclein is involved in the modulation of dopaminergic neurotransmission, these results deliver further pathophysiological explanations of craving mechanisms. PMID- 15897721 TI - Lower packing density of glial fibrillary acidic protein-immunoreactive astrocytes in the prelimbic cortex of alcohol-naive and alcohol-drinking alcohol preferring rats as compared with alcohol-nonpreferring and Wistar rats. AB - BACKGROUND: Low packing density of glial cells, possibly astrocytes, has been described in the prefrontal cortex and hippocampus of "uncomplicated" alcoholics. Astrocytes perform crucial support functions in the processing of neurotransmitters and transfer of energy substrates from blood to cortical neurons. It is still unknown whether attrition in the numbers of astrocytes is only a consequence of prolonged alcohol abuse or also predates the exposure to alcohol in subjects at risk for alcohol dependence. METHODS: We used alcohol preferring (P) rats exposed ad libitum for 2 or 6 months to either water only or 10% ethanol and alcohol-nonpreferring (NP) rats and nonselected Wistar rats exposed only to water for 2 months. Sections through the rat frontal cortex were immunostained for glial fibrillary acidic protein (GFAP), a specific marker of astrocytes. The packing density of GFAP-immunoreactive (IR) astrocytes and the area fraction of GFAP immunoreactivity were measured in the prelimbic cortex (PLC) using the dissector probe and analysis of binary images of GFAP immunostaining, respectively. RESULTS: The packing density of GFAP-IR astrocytes was significantly lower in both alcohol-naive and alcohol-exposed P rats than in NP rats or Wistar rats. The area fraction of GFAP immunoreactivity was significantly lower in the alcohol-exposed P rats than in NP rats, Wistar rats, and alcohol-naive P rats. CONCLUSION: These results suggest that low density of GFAP-IR astrocytes in the PLC of P rats predates the exposure to alcohol and might be a factor contributing to the increased risk for alcohol dependence. In addition, prolonged free-choice alcohol drinking may reduce the extent of GFAP-IR processes in the PLC of P rats. PMID- 15897722 TI - Family history of alcoholism and response to amphetamine: sex differences in the effect of risk. AB - BACKGROUND: Individuals at risk for alcoholism exhibit an enhanced stimulant response to alcohol. It is not known whether individuals at risk also exhibit a heightened sensitivity to other drugs with stimulant properties. METHODS: Healthy young men and women each received, in separate sessions, placebo and 10 mg of d amphetamine in counterbalanced order. Stimulant and sedative subjective effects were recorded before and three times after capsule administration using the Biphasic Alcohol Effects Scale. The sample comprised 19 family-history-positive (FHP; 58% women) and 53 family-history-negative (FHN; 51% women) participants. RESULTS: As compared with placebo, amphetamine increased ratings of stimulation in the sample as a whole. In addition, the ratings revealed an enhanced, as well as a protracted, stimulant response to amphetamine among FHP men, as compared with FHN men: for FHP men, ratings of stimulation made 3 and 6 hr after amphetamine administration were greater than baseline ratings. Moreover, in FHP men, the effect of amphetamine, as compared with placebo, was most evident 6 hr after capsule administration. In contrast, despite a dose x hour interaction in FHN men, post hoc comparisons revealed no differences between the baseline and any of the postamphetamine measurements or between amphetamine and placebo ratings at any of the time points. Among women, the drug effect did not differentiate the family-history groups. CONCLUSIONS: Consistent with previous research on alcohol, high-risk men exhibited a heightened stimulant response to amphetamine. Thus, for men, sensitivity to the stimulant properties of drugs may be an endophenotype for alcoholism. Whereas the present results suggest that women at risk do not exhibit an enhanced stimulant response to amphetamine, further study is needed, including evaluation at various points in the menstrual cycle. PMID- 15897723 TI - Detection of recent ethanol intake with new markers: comparison of fatty acid ethyl esters in serum and of ethyl glucuronide and the ratio of 5 hydroxytryptophol to 5-hydroxyindole acetic acid in urine. AB - BACKGROUND: At present, recent ethanol consumption can be routinely detected with certainty only by direct measurement of ethanol concentration in blood or urine. Because ethanol is rapidly eliminated from the circulation, however, the time span for this detection is in the range of hours. Several new markers have been proposed to extend the detection interval, but their characteristics have not yet justified their use in routine clinical practice. We therefore investigated three new markers and compared their kinetics and sensitivities: (1) fatty acid ethyl esters (FAEEs) in serum, (2) ethyl glucuronide (EtG) in urine, and (3) the ratio of 5-hydroxytryptophol to 5-hydroxyindole acetic acid (5-HTOL/5-HIAA) in urine. METHODS: Seventeen healthy men participated in a drinking experiment. Blood and urine samples were collected twice daily on three consecutive days and once daily on days 4 and 5. Ethanol concentration was determined by gas chromatography, FAEE levels, by gas chromatography with mass spectrometry, EtG concentration, by liquid chromatography-tandem mass spectrometry, and 5-HTOL/5-HIAA ratio, by high performance liquid chromatography. RESULTS: The peak serum ethanol concentrations of the subjects ranged from 5.4 to 44.7 mmol/liter (mean +/- SD, 30.1 +/- 9.1 mmol/liter). In the case of the serum ethanol determination, 100% sensitivity was reached only immediately after the end of the drinking experiment, and in the case of FAEE levels and 5-HTOL/5-HIAA ratio, it tested for 6.7 hr after the end of the ethanol intake. Thereafter, these latter parameters declined until 15.3 hr (FAEEs) and 29.4 hr (5-HTOL/5-HIAA), subsequently remaining in a stable range until 78.5 hr without further decrease. In contrast, EtG concentration showed 100% sensitivity until 39.3 hr and thereafter decreased, falling to below the limit of quantification of 0.1 mg/liter at 102.5 hr. CONCLUSION: After moderate drinking, EtG in the urine proved to be a superior marker of recent ethanol consumption in healthy subjects. This is because EtG is a direct ethanol metabolite, it occurs in the urine only when ethanol has been consumed, and its sensitivity remains at the level of 100% for 39.3 hr. PMID- 15897724 TI - Mean corpuscular volume and ADH1C genotype in white patients with alcohol associated diseases. AB - BACKGROUND: Alcohol abuse is associated with several gastrointestinal diseases, such as esophageal carcinoma, chronic alcoholic pancreatitis, and liver cirrhosis. Increased mean corpuscular volume (MCV) has been recognized as a biomarker for alcohol abuse and heavy drinkers. Recent studies from Japan revealed that macrocytosis is related to ALDH-2/2 genotype, leading to increased acetaldehyde accumulation. It has also demonstrated that increased MCV values could also be an independent biomarker for esophageal cancer in Asians. Therefore, the aim of the current study was to investigate possible associations of MCV value with polymorphisms of ADH1C in white patients with alcohol associated esophageal carcinoma, chronic alcoholic pancreatitis, and alcoholic cirrhosis as well as in heavy drinkers without organ damage. METHODS: In this study, a total of 510 alcoholic patients were enrolled with esophageal cancer (n = 98), chronic pancreatitis (n = 98), alcoholic liver cirrhosis (n = 151), and alcohol abuse without gastrointestinal disease (n = 163). ADH1C genotyping was performed by PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis from whole blood. The relation between MCV and ADH1C gene polymorphisms (ADH1C*1 and 1C*2) controlled for the amount of drinking, smoking, and age were investigated using both univariate and multivariate analysis. RESULTS: In univariate analysis, higher alcohol consumption was associated with increased MCV. Other variables were not associated with macrocytosis. In multiple linear regression analysis, after adjustment for age and smoking, higher alcohol consumption and female sex were independently associated with higher MCV values. No other variables, including which alcohol-associated disease the patient had, had an independent effect. Adding ADH genotype rendered no independent significant effect on MCV value. CONCLUSIONS: In a white population, MCV values were not associated with genotype polymorphisms of ADH1C. In contrast to findings in Asians, macrocytosis does not seem to be an independent biomarker for esophageal cancer. The role of ADH1C polymorphism in increasing MCV and the potential use of MCV as a marker for esophageal carcinoma are still pending. PMID- 15897725 TI - The effectiveness of anxiety treatment on alcohol-dependent patients with a comorbid phobic disorder: a randomized controlled trial. AB - OBJECTIVE: Evidence has emerged which indicates that the post-treatment relapse rate for alcohol-dependent patients with a comorbid anxiety disorder is higher than for alcohol-dependent patients without a comorbid anxiety disorder. The question raised by this evidence is whether the relapse rate in these dually diagnosed patients could be reduced if they were given additional treatment for the comorbid anxiety disorder. We attempted to answer this question by conducting a trial among patients with a double diagnosis of alcohol dependence and agoraphobia or social phobia. METHOD: We conducted a 32-week randomized controlled trial among 96 abstinent patients with a primary diagnosis of alcohol dependence and a comorbid anxiety disorder involving agoraphobia or social phobia. The patients were randomly assigned to an intensive psychosocial relapse prevention program on its own (n = 49) or in combination with an anxiety treatment program comprising cognitive behavioral therapy (CBT) and optional pharmacotherapy consisting of an SSRI (n = 47). The primary outcome measure was the percentage of patients who suffered an alcohol relapse during a 32-week period. The secondary outcome measures were total abstinence, a reduction in the days of heavy drinking, and less severe anxiety symptoms. RESULTS: Although the additional therapy clearly reduced the anxiety symptoms, it had no significant effect on the alcohol relapse rates. CONCLUSION: Anxiety treatment for alcohol dependent patients with a comorbid anxiety disorder can alleviate anxiety symptoms, but it has no significant effect on the outcome of alcohol treatment programs. PMID- 15897726 TI - Chemical dependency and psychiatric services for adolescents in private managed care: implications for outcomes. AB - BACKGROUND: Many adolescents with alcohol and drug problems have mental health comorbidities. The literature suggests that patients entering chemical dependency (CD) treatment with co-occurring problems have less successful outcomes, including treatment dropout and relapse. We examined the impact of psychiatric services on treatment initiation, retention, and alcohol and drug abstinence outcomes for adolescents in CD treatment. METHODS: Participants were 419 adolescents aged 12-18 years who were seeking treatment at four CD programs of a nonprofit, managed care, group model health system and a parent or guardian for each adolescent. We surveyed participants at intake and 6 months and examined clinical and administrative data on diagnoses and CD and psychiatric utilization. Six-month response rates were 91% for adolescents and 93% for parents. RESULTS: Fifty-five percent of the patients with treatment intakes had at least one psychiatric diagnosis in addition to a substance use disorder. Compared with matched controls, patients with CD intakes had higher rates of depression, anxiety, eating disorders, attention deficit hyperactivity disorder, conduct disorder, and conduct disorder including oppositional defiant disorder. Thirty one percent of the full sample had psychiatric visits in the 6 months after intake; among those with a psychiatric diagnosis, 54% had a psychiatric visit. Girls and those with higher Youth Self-Report internalizing scores were more likely to have a psychiatric visit (OR = 2.27, p < 0.001 and OR = 1.05, p < 0.0001, respectively). Adolescents receiving psychiatric services were more likely to be abstinent from both alcohol and drugs than those not receiving these services (OR = 1.57, 95% CI = 0.98-2.5) and more likely to be alcohol abstinent (OR = 1.68, 95% CI = 1.00-2.85). Those adolescents at colocated clinics had higher odds of abstinence from both alcohol and drugs (OR = 1.57, 95% CI = 1.03 2.39) and drugs (OR = 1.84, 95% CI = 1.87-2.85) and of returning after intake to initiate CD treatment (OR = 2.28, 95% CI = 1.44-3.61, p < 0.001) than others. CONCLUSIONS: Our results demonstrate the need for psychiatric treatment of adolescents in CD treatment and highlight the importance of their receiving such services. PMID- 15897727 TI - Patterns of DSM-IV alcohol abuse and dependence criteria among adolescents and adults: results from the 2001 National Household Survey on Drug Abuse. AB - BACKGROUND: Studies of the prevalence of DSM-IV alcohol use disorders (AUDs) in general population samples of adolescents and adults are rare. Comparisons of the prevalence of alcohol abuse and dependence in adolescent surveys with the prevalence in adult general population surveys are confounded by differences in survey design and measurement, thereby reducing their validity. The purpose of the present study was to examine the effects of age, sex, race/ethnicity, and drinking status on the prevalence of DSM-IV alcohol abuse and dependence (including diagnostic orphans) and associated diagnostic criteria among adolescents and adults aged 12-65 years in a single representative sample of the US population. METHODS: This study was based on data from the 2001 National Household Survey on Drug Abuse public use file. Of the 55,561 subjects in the survey, 33,576 (60.5%) reported alcohol use in the past year and provided information on DSM-IV AUD criteria. DSM-IV AUD criteria were assessed by questions related to specific symptoms occurring during the past 12 months. RESULTS: Overall, the most prevalent criteria of DSM-IV alcohol dependence were "tolerance" and "time spent obtaining alcohol, drinking, or getting over its effects." The most prevalent criterion of DSM-IV alcohol abuse was "hazardous use." The prevalence of alcohol abuse only and of dependence with and without abuse was highest among respondents aged 18-23 years, followed by respondents aged 12-17 years, and lowest among respondents aged 50 years and older. Among subgroups of current and heavier drinkers, differences between adolescents and young adults were less pronounced, especially among females. For each age group, the prevalence of alcohol abuse only was greater than the prevalence of dependence (with or without abuse). The abuse-to-dependence ratios also were generally consistent across age groups and slightly higher among males (2.1:1.0) than females (1.6:1.0). CONCLUSIONS: The higher prevalence for some dependence criteria among adolescents and young adults as measured in the present study may blur the distinction between symptom reports associated with the normative development of drinking patterns and clinically relevant aspects of DSM-IV alcohol dependence. PMID- 15897728 TI - Reinforcing mood effects of alcohol in coping and enhancement motivated drinkers. AB - BACKGROUND: People may be motivated to drink because of differential sensitivities to the rewarding outcomes of alcohol consumption. Previous research has demonstrated that alcohol may produce both potentially positive reinforcing effects (i.e., increased elation; Conrad et al., 2001) and potentially negative reinforcing effects (i.e., anxiolytic effects; Levenson st al., 1980). It was desired to test the effects of alcohol on mood in a sample of two groups of drinkers that report different motivations for alcohol use. It was hypothesized that both potentially positive and negative reinforcing mood effects would be observed and that these effects would be moderated by drinking motive. METHODS: Twenty-four drinkers with Coping Motives (CMs) and 24 drinkers with Enhancement Motives (EMs) were randomly assigned to either an alcohol condition (target blood alcohol level of 0.08%) or a placebo condition. Participants used the Profile of Mood States-Bipolar (Lorr, 1983) to report their mood at (1) sober baseline, (2) after beverage consumption, and (3) during anticipation of a self-disclosing speech (a stressor). RESULTS: As hypothesized, after drinking, those in the alcohol group reported increased feelings of elation and energy relative to sober baseline. Those receiving alcohol also reported feeling more confused and anxious after beverage consumption. Also as hypothesized, participants receiving alcohol reported feeling increased sedation during anticipation of the stressor, whereas those receiving placebo reported increased energy during this period. Contrary to the hypothesis, none of these effects were moderated by drinking motive. CONCLUSIONS: Although potentially positive and negative reinforcing mood effects of alcohol were observed, CM and EM drinkers were not differentially sensitive to these effects. However, it is possible that EM drinkers may highly value the baseline stimulating effects of alcohol, whereas CM drinkers may highly value the anxiolytic effects that were observed during anticipation of the stressor. PMID- 15897729 TI - Alcohol use and sexual risk behavior among human immunodeficiency virus-positive persons. AB - BACKGROUND: This study was undertaken to determine if alcohol use is associated with sexual risk taking among human immunodeficiency virus (HIV)-infected persons. METHODS: Cross-sectional interviews of 262 HIV-infected patients in the Brown University AIDS Program were performed. Factors associated with any sexual activity, unsafe sexual activity, and a 4-fold typology of sexual risk were examined. Alcohol measures included drinking days, drinks per drinking day, binge drinking, and hazardous alcohol use. RESULTS: The sample was 58% male and 40% white; 67% of patients were self-identified as heterosexual, and 48% drank alcohol. Nearly two thirds of patients reported sexual activity in the past 6 months, with 38% reporting unprotected sex during that period. All measures of alcohol use were significantly associated with any sexual activity and with unsafe sexual behavior. As an example, controlling for age, HIV transmission risk, marital status, and HIV clinical indicators, hazardous drinkers were 5.64 times more likely to report unprotected sex and have multiple partners (p < 0.01) than were those not drinking at hazardous levels. CONCLUSIONS: A high proportion of HIV-infected persons were sexually active and having unsafe sex. Alcohol, at all levels of use, was associated with increased sexual risk taking. PMID- 15897730 TI - Effectiveness of the derived Alcohol Use Disorders Identification Test (AUDIT-C) in screening for alcohol use disorders and risk drinking in the US general population. AB - BACKGROUND: The three consumption questions from the Alcohol Use Disorders Identification Test (AUDIT-C) are increasingly used as a screener for alcohol use disorders (AUDs) and risk drinking. METHODS: In a representative sample of US adults 18 years of age and older, AUDIT-C scores (derived from consumption questions embedded in a large national survey) were used to estimate sensitivity, specificity, and areas under receiver operator characteristic curves (AUROCs) for alcohol dependence, any AUD, and risk drinking. AUDs were defined according to DSM-IV criteria. For men, risk drinking was defined as consuming >14 drinks per week or >4 drinks in a single day at least once a month; for women, the weekly and daily limits were >7 drinks and >3 drinks, respectively. The derived AUDIT-C was evaluated among past-year drinkers (n = 26,946), within the total population (n = 43,093), in groups defined by age, sex, and race/ethnicity, and among pregnant women, persons attending an emergency room, and college students. RESULTS: For past-year drinkers, the AUROCs for the derived AUDIT-C were 0.887 for alcohol dependence, 0.860 for any AUD, and 0.966 for risk drinking. Scores were higher in the total population, 0.931, 0.917, and 0.981, respectively. The derived AUDIT-C performed slightly better in screening for dependence among women than men. Screening for risk drinking was better among men, probably because the third AUDIT-C question directly mirrors one of the definitions of risk drinking for men but not for women. Performance in pregnant women, past-year emergency room patients, and college students was on a par with performance in the general population. CONCLUSIONS: The derived AUDIT-C performs well in screening for AUDs and risk drinking. The use of variable cut points for men and women improves its sensitivity and specificity. Validation in a realistic screening situation, in which the AUDIT-C questions are asked as stand-alone and not embedded items, is a critical future step. PMID- 15897731 TI - Differential teratogenic effect of alcohol on embryonic development between C57BL/6 and DBA/2 mice: a new view. AB - BACKGROUND: Alcohol exposure during the fetal stage generates variable severity in different organs, as seen in fetal alcohol syndrome and fetal alcohol effect. Whether genetic factors or conditions of alcohol exposure influence the susceptibility to alcohol-related developmental impairment remains a question. METHODS: To investigate the contribution of genotype to the susceptibility to alcohol-induced toxicity during development beyond confounding maternal factors and variables of alcohol exposures, the authors tested the effect of alcohol exposure under definitive concentration using a whole embryonic culture of two inbred strains previously known to be vulnerable (C57BL/6 [C6]) or resistant (DBA/2 [D2]) to alcohol. On gestational day 8, embryos from each group bearing three to six somites were collected and then cultured for 44 hr in a medium added with 400 mg/dl of ethanol. The viability and morphological malformations, as well as developmental staging of the embryos, were all scored at the end of the culture. RESULTS: The authors found, in contrast to previous reports, that alcohol treatment retarded embryonic growth and induced abnormalities, including the neural tube opening and the hypoplasia of the optic vesicle in both strains. However, alcohol specifically compromised the heart and caudal neural tube in C6, whereas it specifically decreased the number of somites and the development of branchial bars among others in D2. CONCLUSIONS: These results demonstrated that both strains of embryos are vulnerable to the same amount and pattern of alcohol exposures at the same developmental stage, but each with unique vulnerability in specific organs, with alcohol having greater teratogenic effects in D2 than in C6. These differential vulnerabilities are results of greater genetic influence, rather than the maternal influence or conditions of alcohol. PMID- 15897732 TI - Myocardial antioxidant status in chronic alcoholism. AB - BACKGROUND: Excessive ethanol intake is one of the most frequent causes of acquired dilated cardiomyopathy in developed countries. The pathogenesis is multifactorial, with the antioxidant imbalance of cardiac muscle being a potential factor. The current study evaluates myocardial antioxidant status in ethanol consumers and its relation to cardiac damage. METHODS: The authors assessed superoxide dismutase, glutathione peroxidase, and glutathione reductase enzyme activities as well as the total antioxidant status capacity in myocardial samples obtained from organ donors with sudden death of traumatic or neurological origin. They studied 23 high-dose chronic alcohol consumers, 27 individuals with long-standing hypertension, and 11 healthy controls. Cardiomyopathy was defined according to standard functional and histological criteria. RESULTS: Patients with dilated cardiomyopathy, either of alcoholic or hypertensive origin, showed increased myocardial superoxide dismutase activities compared with patients without cardiomyopathy (p < 0.001, both) and controls (p < 0.05, both). Total antioxidant status capacity and the activity of glutathione peroxidase and glutathione reductase enzymes were similar in all groups. Superoxide dismutase activity was related to the presence of cardiac enlargement and the degree of cardiac histological damage. The amount and type of alcoholic beverages as well as the nutritional status of the patients were not related to myocardial antioxidant activity. CONCLUSIONS: The presence of dilated cardiomyopathy, of either alcoholic or hypertensive origin, is related to an increase in myocardial superoxide dismutase activity. PMID- 15897733 TI - Corticosterone increases damage and cytosolic calcium accumulation associated with ethanol withdrawal in rat hippocampal slice cultures. AB - BACKGROUND: Evidence suggests that stress hormones (i.e., glucocorticoids) may be increased during acute or chronic consumption of ethanol and during withdrawal from ethanol consumption, effects that may contribute to the development of cognitive impairment. The goal of the current studies was to examine the hypothesis that increased glucocorticoid levels in conjunction with ethanol exposure and withdrawal may cause hippocampal damage. METHODS: Organotypic hippocampal slice cultures were exposed to 50 mM ethanol for 10 days and withdrawn for 1 day. After withdrawal, cytotoxicity and cytosolic Ca2+ accumulation were measured using the nucleic acid stain propidium iodide and Calcium Orange, AM, respectively. Cultures were also treated with nontoxic concentrations of corticosterone (0.001-1 microM) during ethanol exposure and withdrawal or only during withdrawal. Additional cultures were coexposed to corticosterone and RU486 (0.1-10.0 microM), spironolactone (0.1-10.0 microM), or MK-801 (20 microM) during ethanol exposure and/or withdrawal. RESULTS: Ethanol withdrawal did not increase propidium iodide fluorescence and cytosolic Ca2+ levels. However, significant increases in propidium iodide fluorescence and in cytosolic Ca2+ accumulation were observed in cultures when corticosterone (> or = 100 nM) was exposed during ethanol treatment and/or withdrawal. These effects of corticosterone on ethanol withdrawal were attenuated by RU486 and MK-801 but not by spironolactone coexposure. CONCLUSIONS: This report demonstrated that corticosterone exposure during ethanol treatment and/or withdrawal resulted in significant hippocampal damage, possibly via activation of glucocorticoid receptors and enhancement of the glutamatergic cascade. The findings from these studies suggest that glucocorticoids contribute to the neuropathological consequences of alcohol dependence in humans. PMID- 15897734 TI - Smoke exposure exacerbates an ethanol-induced defect in mucociliary clearance of Streptococcus pneumoniae. AB - BACKGROUND: Alcoholics and smokers are particularly susceptible to pulmonary infections caused by Streptococcus pneumoniae, the pneumococcus. Infection begins when pneumococci colonizing the nasopharynx are aspirated into the lower respiratory tract. The major host defense against this movement is the mucociliary clearance apparatus. Both cigarette smoke and ethanol (EtOH) exposure alter ciliary beating and protein kinase activity in the respiratory mucosa in vitro, but their effects on bacterial clearance in the intact animal have not been determined. METHODS: Male Sprague Dawley rats were exposed twice daily for 12 weeks to either the smoke generated from 30 cigarettes (smoke-exposed) or room air (sham-exposed). For the last five weeks of smoke exposure, the rats were fed Lieber-DeCarli liquid diets containing 0%, 16%, 26%, or 36% EtOH calories. The rats then were infected intranasally with S. pneumoniae, and movement of the organisms into the lower respiratory tract was quantified by plate counts of the tracheas and lungs 4 hr later. Ciliary beat frequency (CBF) analysis was performed on tracheal ring explants from each animal before and after stimulation with the beta-agonist isoproterenol, and tracheal epithelial cell protein kinase C (PKC) activity was measured. RESULTS: Ingestion of any of the EtOH-containing diets resulted in a dose-dependent increase in movement of S. pneumoniae into the rats' lungs. This EtOH-induced defect was augmented further by concurrent smoke exposure, although smoke exposure alone had little effect on S. pneumoniae movement. Smoke, but not EtOH exposure, activated tracheal epithelial cell PKC. Increased movement of organisms into lungs correlated with a decrease in CBF and loss of the ciliary response to isoproterenol. CONCLUSION: EtOH ingestion in our model facilitated movement of S. pneumoniae into rats' lungs, a phenomenon exacerbated by concurrent smoke exposure. Furthermore, the organism's movement into the lungs correlated with a blunting of the rats' ciliary response to an established stimulus. Defects in mucociliary clearance thus may be one cause of the increased risk of pneumococcal infections in people who abuse alcohol, particularly if they also smoke. PMID- 15897735 TI - Effects of light and dark beer on hepatic cytochrome P-450 expression in male rats receiving alcoholic beverages as part of total enteral nutrition. AB - BACKGROUND: Alcoholic beverages contain many congeners in addition to ethanol. Therefore, consumption of alcoholic beverages may have considerably different effects on expression of hepatic microsomal monooxygenases than the relatively selective induction of cytochrome P-450 (CYP) 2E1 observed after consumption of pure ethanol. METHODS: : In the current study, we compared the effects of two beers: lager (a light roasted beer) and stout (a dark roasted beer) with those of an equivalent amount of pure ethanol on hepatic CYP expression. Beer or pure ethanol was part of a complete total enteral nutrition diet that was infused intragastrically into male Sprague Dawley rats for 21 days. At the end of the infusion period, rats were euthanized, and liver and intestinal microsomes were prepared. Expression and activity of CYP1A1/2, CYP2B1, CYP2E1, CYP3A, and CYP4A were assessed by Western immunoblotting and by using CYP enzyme-specific substrates, respectively. RESULTS: mRNA and protein levels of CYP4A1 were elevated only in stout-treated animals. However, lauric acid 12-hydroxylase activity (a CYP4A-specific activity) was reduced (p < or = 0.05) in microsomes from lager- and stout-fed rats. After oxidation with potassium ferricyanide, this activity was significantly increased in microsomes from stout-fed animals. The relative expression of CYP2E1 and CYP2B1 and the activities toward p-nitrophenol, pentoxyresorufin, or benzyloxyresorufin did not differ between beers or compared with pure ethanol or controls. However, the mean expression of CYP1A2, CYP3A, and CYP4A apoproteins was greater in liver microsomes from stout-infused rats than in those from lager-infused rats, ethanol-infused rats, and diet controls (p < or = 0.05). In addition, although no significant differences were observed in ethoxyresorufin O-dealkylase (EROD), methoxyresorufin O-dealkylase (MROD), midazolam, or testosterone hydroxylase activities between groups, stout-infused rats had greater hepatic microsomal erythromycin N-demethylase activity than other groups (p < or = 0.05). CONCLUSIONS: Stout contains components other than ethanol that interact in a complex fashion with the monooxygenase system. PMID- 15897736 TI - Neuroimaging of gender differences in alcohol dependence: are women more vulnerable? AB - BACKGROUND: Alcoholic brain damage has been demonstrated in numerous studies using neuropathology and brain imaging techniques. However, gender differences were addressed only in a few studies. Recent research has shown that development, course, and consequences of alcohol dependence may differ between female and male patients. Our investigation was built upon earlier research where we hypothesized that women develop alcoholic brain damage more readily than men do. To further compare the impact of alcohol dependence between men and women, we examined brain atrophy in female and male alcoholics by means of computed tomography (CT). METHODS: The study group consisted of a total of 158 subjects (76 women: 42 patients, 34 healthy controls; 82 age-matched men: 34 patients, 48 healthy controls). All patients had a DSM-IV and ICD-10 diagnosis of alcohol dependence. CT with digital volumetry was performed twice in patients (at the beginning and end of the 6-week inpatient treatment program) and once in controls. RESULTS: Patients of both genders had consumed alcohol very heavily. Although the average alcohol consumption in the year before the study was significantly lower in female alcoholics, this gender difference disappeared when controlled for weight. However, women had a significantly shorter duration of alcohol dependence. Despite this fact, both genders developed brain atrophy to a comparable extent. Brain atrophy was reversible in part after 6 weeks of treatment; it did not reach the level in the control groups. CONCLUSIONS: Gender-specific differences in the onset of alcohol dependence were confirmed. This is in line with the telescoping effect, where a later onset and a more rapid development of dependence in women were described. Under the assumption of a gradual development of consequential organ damage, brain atrophy seems to develop faster in women. As shown in other organs (i.e., heart, muscle, liver), this may confirm a higher vulnerability to alcohol among women. PMID- 15897737 TI - Quantifying the risks associated with exceeding recommended drinking limits. AB - BACKGROUND: Although daily and weekly drinking limits demonstrate strong sensitivity and specificity in identifying alcohol use disorders (AUDs), there are no descriptive data that present the risks associated with exceeding these limits in a format suitable for presentation to patients, students, and the general public. METHODS: Data collected in the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions were used to estimate the risks of past year DSM-IV alcohol abuse and dependence associated with various frequencies of exceeding daily drinking limits (no more than 4 drinks for men; no more than 3 drinks for women) in a nationally representative sample of 26,946 US drinkers 18 years of age and older. These risks were further categorized by whether weekly drinking limits (no more than 14 drinks for men; no more than 7 drinks for women) were exceeded and by maximum number of drinks consumed in the past year. RESULTS: The prevalence of alcohol dependence with abuse increased in a fairly linear fashion with frequency of exceeding daily drinking limits. The prevalence of dependence alone (no abuse) and abuse alone (no dependence) peaked among persons who exceeded the daily limits twice a week and then leveled off, because individuals became increasingly likely to have both disorders at higher frequencies. Exceeding the weekly limits generally increased the risks of both disorders after accounting for frequency of exceeding the daily limits, but not always to a significant extent. Likewise, maximum quantity of drinks consumed was positively associated with the risks of AUDs even after accounting for frequency of risk drinking. There were few gender differences in the risk of dependence after adjusting for frequency of exceeding daily drinking limits, but the risk of alcohol abuse remained greater among men. CONCLUSIONS: These data provide a useful tool for illustrating the broad range of risk of AUDs associated with exceeding recommended drinking limits, and they support the utility of the daily and weekly drinking limits in predicting AUDs. PMID- 15897738 TI - Ovarian serous tumors of low malignant potential (borderline tumors): outcome based study of 276 patients with long-term (> or =5-year) follow-up. AB - The natural history, classification, and nomenclature of ovarian serous tumors of low malignant potential (S-LMP) (serous tumors of borderline malignancy, atypical proliferating tumors) are controversial. To determine long-term outcome for patients with S-LMP and further evaluate whether S-LMP can be stratified into clinically benign and malignant groups, the clinicopathologic features of 276 patients with S-LMP and > or =5 year follow-up were studied. The histology of the ovarian primary, extraovarian implants, and recurrent tumor(s) were characterized using World Health Organization criteria and correlated with FIGO stage and clinical follow-up. After censoring nontumor deaths, overall survival and disease free survival for the 276 patients was 95% (98% FIGO stage I; 91% FIGO II-IV) and 78% (87% FIGO stage I; 65% FIGO stage II-IV), respectively. Unresectable disease (P < 0.001) and invasive implants (P < 0.001) were associated with decreased survival. When compared with typical S-LMP, S-LMP with micropapillary features were more strongly associated with invasive implants (P < 0.008) and decreased overall survival (P = 0.004), but patient outcome with micropapillary S-LMP was not independent of implant type. Stromal microinvasion in the primary tumor was also correlated with adverse outcome, independent of stage of disease, micropapillary architecture, and implant type (P = 0.03). There was no association between outcome and lymph node status. Transformation to low-grade serous carcinoma occurred in 6.8% of patients at intervals of 7 to 288 months (58% > or = 60 months) and was strongly associated with increased tempo of disease and decreased survival (P < 0.001). S-LMP forms a heterogeneous group, morphologically and clinically distinct from benign serous tumors and serous carcinoma. The majority of S-LMP are clinically benign, but recurrences are not uncommon, and persistent disease as well as deaths occur. Progression to low grade serous carcinoma is highly predictive of more aggressive disease. Other features associated with recurrent and/or progressive disease include FIGO stage, invasive implants, microinvasion in the primary tumor, and micropapillary architecture. These predictors tend to co-occur, and no single clinical or pathologic feature or combination of features identify all adverse outcomes. The small, but significant risk of progression over time to low-grade serous carcinoma emphasizes the need for prolonged follow-up in patients with S-LMP. PMID- 15897739 TI - A proposal for a new and more practical grading scheme for pancreatic ductal adenocarcinoma. AB - There is no uniformly applied grading system for pancreatic ductal adenocarcinoma (DA). The scheme advocated by the WHO is essentially that of Kloppel et al, and is based on the "highest grade" focus. Although it is precise with good prognostic value, it is unfortunately not widely applied, largely because of the lack of recognition and partly because of its complex nature (interpretation of multiple parameters). Furthermore, it is fundamentally different from the one used in Japan, which evaluates the overall pattern. To establish a more widely applicable, practical, and clinically relevant grading system, a scheme similar to Gleason's scoring system was developed and tested on 112 cases of resected pancreatic DA and was compared with the WHO system. In the grading system devised, patterns (P) of infiltration were classified as follows: P1, well defined glands with easily discernible contours; P2, fused or poorly formed glands with ill-defined contours; P3, nonglandular patterns. A score was then obtained by the summation of the predominant and the secondary patterns. Scores < or =3 (at least some well-formed glands and no nonglandular pattern) was graded as G1, 4 as G2, and > or =5 (at least some nonglandular patterns and no well formed glandular pattern) as G3. Seventy-three percent of the cases displayed mixed patterns, with disparate patterns (P1 with P3) in 13%, confirming the high degree of heterogeneity of DA. There was a significant correlation between grade and survival, better than the correlation between survival and either the major or minor patterns evaluated separately. The median survival for G1, G2, and G3 were 22, 14, and 8 months; 1-year survival 68%, 44%, and 33%; 2-year was 67%, 11%, and 0%; and 3-year was 23%, 4%, and 0%, respectively (P = 0.0019). In a multivariate analysis, correlating survival with grade, tumor size, and lymph node status, the grade was the strongest independent predictor of survival. Odds ratio of dying of disease were 3.56 (P < 0.0001) in G3 versus G1, 1.79 (P = 0.058) in G2 versus G1, and 1.98 (P = 0.03) in G3 versus G2. Compared with this, the same odds ratio were 1.17 (P = 0.01) in tumors >2 cm versus < or =2 cm and 1.78 (P = 0.01) in cases with positive versus negative lymph nodes. The WHO grading scheme was not found to have as good a correlation with survival in this study, with WHO grade 2 showing a better survival than 1. The reproducibility of both the proposed grading system and that of WHO were found to be moderately good (with kappa values of 0.43 and 0.44, respectively), when 32 slides of DA were graded by four independent observers. The grading scheme for pancreatobiliary adenocarcinoma proposed here is highly applicable because it is practical and readily adoptable. It reflects biologic characteristics of ductal carcinoma (prominent tubule formation and tumor heterogeneity). Most importantly, it is clinically relevant with good prognostic value. Lastly, it is also applicable for use in research, by utilizing "patterns," even in small specimens like microarrays or biopsies. PMID- 15897740 TI - Columnar cell lesions of the breast: the missing link in breast cancer progression? A morphological and molecular analysis. AB - Columnar cell lesions (CCLs) of the breast are a spectrum of lesions that have posed difficulties to pathologists for many years, prompting discussion concerning their biologic and clinical significance. We present a study of CCL in context with hyperplasia of usual type (HUT) and the more advanced lesions ductal carcinoma in situ (DCIS) and invasive ductal carcinoma. A total of 81 lesions from 18 patients were subjected to a comprehensive morphologic review based upon a modified version of Schnitt's classification system for CCL, immunophenotypic analysis (estrogen receptor [ER], progesterone receptor [PgR], Her2/neu, cytokeratin 5/6 [CK5/6], cytokeratin 14 [CK14], E-cadherin, p53) and for the first time, a whole genome molecular analysis by comparative genomic hybridization. Multiple CCLs from 3 patients were studied in particular detail, with topographic information and/or showing a morphologic spectrum of CCL within individual terminal duct lobular units. CCLs were ER and PgR positive, CK5/6 and CK14 negative, exhibit low numbers of genetic alterations and recurrent 16q loss, features that are similar to those of low grade in situ and invasive carcinoma. The molecular genetic profiles closely reflect the degree of proliferation and atypia in CCL, indicating some of these lesions represent both a morphologic and molecular continuum. In addition, overlapping chromosomal alterations between CCL and more advanced lesions within individual terminal duct lobular units suggest a commonality in molecular evolution. These data further support the hypothesis that CCLs are a nonobligate, intermediary step in the development of some forms of low grade in situ and invasive carcinoma. PMID- 15897741 TI - Distribution of cytokeratins and vimentin in adult renal neoplasms and normal renal tissue: potential utility of a cytokeratin antibody panel in the differential diagnosis of renal tumors. AB - Adult renal epithelial neoplasms (RENs) comprise several distinct clinicopathologic entities with potential prognostic and therapeutic differences. Individual cases can show overlapping morphologic features, necessitating the use of ancillary methods. The purpose of this study was to determine the diagnostic utility of cytokeratin (CK) subtype expression pattern in a wide range of adult RENs. RENs (including clear cell [conventional] renal cell carcinoma (RCC), papillary RCC, chromophobe RCC, renal oncocytoma, collecting duct carcinoma (CDC), renal medullary carcinoma (RMC), urothelial carcinoma, metanephric adenoma (MA), tubulocystic carcinoma (TC) (also known as low-grade collecting duct carcinoma), and mucinous tubular and spindle cell carcinoma) were immunostained for CK subtypes (CK5/CK6, 7, 8, 13, 14, 17, 18, 19, 20), high molecular weight CKs 1, 5, 10, 14 (HMWCK), and vimentin (Vim). The expression pattern of normal kidney was also examined and correlated with RENs. Although there is some overlap, subtypes of RENs show distinctive CK expression profiles that may be useful in several differential diagnostic settings. Clear cell RCCs typically showed a restricted expression pattern of CK8, CK18 and Vim. Papillary RCCs typically expressed CK7, CK8, CK18, CK19, and Vim and could be distinguished from MA (CK7-). Chromophobe RCCs were typically CK7+, CK8+, CK18+, and Vim-, and could be distinguished from oncocytomas (typically CK7-). In oncocytomas, nonspecific staining of unblocked endogenous biotin is a potentially significant diagnostic pitfall. CDC, RMC, and TC demonstrated similar CK expression profiles (with the exception of HMWCK expression limited to CDC), supporting a close relationship between these entities. A panel of CK5/CK6, CK17, and Vim may be helpful in distinguishing CDC (typically CK5/CK6-, CK17-, Vim+) and urothelial carcinoma (typically CK5/CK6+, CK17+, Vim-). In conclusion, CK expression patterns may be helpful in several differential diagnostic situations when dealing with adult RENs. PMID- 15897742 TI - Gastrointestinal stromal tumors of neurofibromatosis type I (von Recklinghausen's disease). AB - Gastrointestinal stromal tumor (GIST), as well as the hyperplastic lesions of intestinal neural tissue and its supporting structures, is a gastrointestinal complication of type 1 neurofibromatosis (NF1) (von Recklinghausen's disease). In the present study, we analyzed the histologic and immunohistochemical features, and the c-kit and PDGFRA gene mutations of 36 GISTs derived from 9 NF1 patients. Distinctively, multiple GISTs arose preferentially in the small intestine. The histologic features of NF1-associated GISTs are almost similar to those of non NF1 GISTs, but characteristically most of the NF1-associated GISTs contained skeinoid fibers. Thirty-three GISTs (92%) showed immunoreactivity for KIT, and 23 tumors (64%) showed diffuse or mosaic-like immunoreactivity for S-100 protein. Hyperplasic lesions, which may be the hyperplasia of interstitial cells of Cajal, were observed around some GISTs. Exons 9, 11, 13, and 17 of the c-kit gene and exons 12 and 18 of the PDGFRA gene were amplified and directly sequenced. Point mutations of c-kit gene or PDGFRA gene were identified only in three (8%) and two (6%) tumors, respectively. NF1-associated GISTs, showing the dual differentiation of interstitial cells of Cajal and Schwann cells, develop in close association with the myenteric nerve structure of gastrointestinal tract of NF1 patients. The point mutations of c-kit and PDGFRA gene may play a limited role in the tumorigenesis of NF1-associated GISTs. PMID- 15897743 TI - Mucinous carcinoma of the skin, primary, and secondary: a clinicopathologic study of 63 cases with emphasis on the morphologic spectrum of primary cutaneous forms: homologies with mucinous lesions in the breast. AB - We present the largest series of mucinous carcinoma involving the skin, describing the histopathologic, immunohistochemical, electron microscopic, and cytogenetic findings. Our aim was fully to characterize the clinicopathologic spectrum and compare it with that seen in the breast. In addition, we wished to reevaluate the differential diagnostic criteria for distinguishing primary mucinous carcinomas from histologically similar neoplasms involving the skin secondarily, and study some aspects of their pathogenesis. We demonstrate that primary cutaneous mucinous carcinomas span a morphologic spectrum compatible to their mammary counterparts. Both pure and mixed types can be delineated morphologically, and some lesions have mucocele-like configurations. Most lesions seem to originate from in situ lesions that may represent, using mammary pathology terminology, ductal hyperplasia, atypical ductal hyperplasia, or ductal carcinoma in situ or a combination of the three. Inverse cell polarity appears to facilitate the progression of the changes similar to lesions in the breast. The presence of an in situ component defines the neoplasm as primary cutaneous, but its absence does not exclude the diagnosis; although for such neoplasms, full clinical assessment is essential. Mammary mucinous carcinoma involving the skin: all patients presented with lesions on chest wall, breast, axilla, and these locations can serve as clue to the breast origin. Microscopically, cutaneous lesions were of both pure and mixed type, and this correlated with the primary in the breast. Dirty necrosis was a constant histologic finding in intestine mucinous carcinomas involving the skin, and this feature may serve as a clue to an intestinal origin. PMID- 15897744 TI - Abundant IgG4-positive plasma cell infiltration characterizes chronic sclerosing sialadenitis (Kuttner's tumor). AB - Chronic sclerosing sialadenitis (CSS) is a cryptogenic tumor-like condition of the salivary gland(s). While immune-mediated processes are suspected in its pathogenesis, and CSS is occasionally reported to be associated with sclerosing pancreatitis, an IgG4-related disease, the exact immunopathologic processes of CSS remain speculative. In this study, we examined the clinicopathologic findings of CSS (12 cases) in comparison with sialolithiasis (8 cases) and Sjogren's syndrome (13 cases), and tried to clarify whether CSS is an IgG4-related disease or not. Submandibular gland(s) were affected in all cases of CSS. CSS cases could be divided into two types: 5 cases were associated with sclerosing lesions in extrasalivary glandular tissue (systemic type), while only salivary gland(s) were affected in the remaining 7 cases (localized type). In the former type, which showed male predominance, bilateral salivary glands were frequently affected, and eosinophilia and elevations of gamma-globulin and IgG in serum were frequently found. Histologically, all cases of CSS showed marked lymphoplasmacytic infiltration admixed with fibrosis and the destruction of glandular lobules. Obliterative phlebitis was found in the affected salivary glands in all cases of CSS. Immunohistochemically, the proportion of IgG4/IgG-positive plasma cells was more than 45% in CSS, while it was less than 5% in controls. The resemblance of the clinicopathologic features of CSS with those of sclerosing pancreatitis suggests the participation of a similar immunopathologic process with IgG4 disturbance in CSS. The abundance of IgG4-positive plasma cells in the lesions would be useful for distinguishing CSS from other forms of sialadenitis. PMID- 15897745 TI - Potential diagnostic use of p16INK4A, a new marker that correlates with dysplasia in oral squamoproliferative lesions. AB - Protein products of tumor suppressor genes are often involved in regulating the cell cycle, and aberrant expression can correlate with underlying genetic mutations. Mutations in the p16 gene have been detected at relatively high rates in squamous cell carcinomas of the oral cavity. However, immunohistochemical staining for the protein product has not been examined as a diagnostic tool for identifying dysplastic lesions in the oral cavity. Sixty cases of biopsies of reactive, inflammatory, and dysplastic lesions of all grades were stained with an antibody to p16INK4A and analyzed for which layer of the epithelium had positive cells. Staining was seen only in the basal or lower third in keratoses and mild dysplasias and was seen in the mid and upper thirds in moderate to severe dysplasia. The staining across larger fragments of biopsied epithelium highlighted skip lesions, with strong staining restricted to dysplastic regions. Inflammatory lesions, including chronic ulcers, showed absent or minimal basal layer staining with p16INK4A. In this preliminary study, p16INK4A shows promise as to a potential marker to aid in recognizing the presence of dysplasia in squamous mucosa of the head and neck, particularly in subtle lesions, and in an inflammatory or ulcerated background. PMID- 15897746 TI - Pancreatic solid and cystic hamartoma in adults: characterization of a new tumorous lesion. AB - Nonneoplastic tumor-like lesions ("pseudotumors") of the pancreas include cystic and noncystic varieties. We report on a solid and cystic tumor-like lesion of the pancreas that occurred in 2 adult patients. The lesions, located in the head and neck of the gland, respectively, were well demarcated and composed of cystic ductal structures embedded in focally inflamed stromal tissue. In addition, one of the lesions showed irregularly arranged but well-differentiated acini and small intralobular and interlobular ducts embedded in hypocellular, fibrotic tissue. Discrete islets were lacking, but immunohistochemical staining for chromogranin A revealed individual scattered endocrine cells evenly distributed between acinar and ductal cells. The surrounding pancreatic parenchyma did not show significant chronic pancreatitis. After tumor removal, the follow-up of the patients was uneventful. Because of the irregular arrangement of otherwise mature tissue components of the pancreas, the lesions were considered solid and cystic hamartomas. Their pathogenesis is so far unknown. PMID- 15897747 TI - A preliminary diagnosis service provides prospective blinded dual-review of all general surgical pathology cases in an academic practice. AB - Quality assurance of diagnostic accuracy in surgical pathology is an important part of a pathologist's total quality management program. At our academic institution, the quality of diagnostic accuracy is monitored via dual-review of every general surgical pathology case, which accounts for nearly 20,000 cases per year. This comprehensive dual-review is achieved by operating a preliminary diagnosis service, staffed by a senior or board eligible resident. Analysis of a portion of our dual-review data (6300 cases) demonstrates an overall diagnostic concordance rate of 95.4% and a clinical major discrepancy rate of 0.29% between the preliminary diagnosis and staff pathologist diagnosis, comparable to other published rates. The incorporation of a preliminary diagnosis service into our academic surgical pathology practice has proven to be beneficial with regard to quality assurance and resident education. Other academic institutions may similarly benefit from the addition of such a service. PMID- 15897748 TI - Expression of membrane-bound mucins (MUC1 and MUC4) and secreted mucins (MUC2, MUC5AC, MUC5B, MUC6 and MUC7) in mucoepidermoid carcinomas of salivary glands. AB - Mucins are glycoproteins normally synthesized by a variety of secretory epithelial cells. The aim of this study was to investigate the expression of mucins (MUC1, MUC2, MUC4, MUC5AC, MUCB, MUC6, MUC7) in mucoepidermoid carcinomas, the most frequent malignant tumor of salivary glands. Forty mucoepidermoid carcinomas and twenty-two normal salivary glands were studied for these mucins by immunohistochemistry from formalin-fixed and paraffin-embedded material. Normal salivary glands frequently expressed MUC1 and MUC4, mainly in ductal cells; MUC5B and MUC7 stained mucous and serous acini respectively of submandibular and minor salivary glands; and MUC5AC and MUC2 were poorly detected in excretory ducts. All mucoepidermoid carcinomas expressed MUC1, and 38/40 tumors expressed MUC4. Both membrane-bound mucins stained membranes and cytoplasm of all cell types (epidermoid, intermediate, mucous, clear and columnar). MUC5AC and MUC5B stained glandular differentiated cells in most tumors (29/40 and 33/40 cases, respectively). MUC6 was positive in 13/40 tumors, and both MUC2 and MUC7 in only 2/40 tumors. The high expression of MUC1 was related to high histologic grades, high recurrence and metastasis rates and a shorter disease-free interval (P < 0.05). Conversely, MUC4 high expression was mainly related to low-grade tumors, lower recurrence rates and a longer disease-free interval (P < 0.05). In conclusion, mucoepidermoid carcinomas of salivary glands usually express MUC1, MUC4, MUC5AC and MUC5B; less frequently MUC6; and rarely MUC2 and MUC7. This mucin expression pattern can be useful for diagnostic purposes. Therefore, MUC1 expression is related to tumor progression and worse prognosis, whereas MUC4 expression is related to a better prognosis. PMID- 15897749 TI - The pathology of breast biopsy site marking devices. AB - This report presents our experience with 54 cases of patients who had excision of breast lesions after deployment of radiographic biopsy site markers at the time of stereotactic biopsy. These were of two types: pellets of a resorbable copolymer of polylactic acid/polyglycolic acid (31 cases) and plugs of bovine collagen (23 cases), both containing embedded metallic clips for long-term radiographic marking. On gross examination, the pellets have a characteristic appearance similar to a soft grain of rice, whereas the collagen plugs are spongiform with variable hemorrhagic changes. Microscopically, there are distinct differences in the morphologic features of these two types of biopsy site markers and the associated tissue reactions. With the pellets, there is an initial cell poor fibrotic reaction around empty spaces followed by a multinucleate giant cell reaction and penetration of the marker core by eosinophilic fibrinous material. The collagen plugs are recognized as eosinophilic, hyalinized, acellular material, accompanied by an inflammatory infiltrate composed predominantly of lymphocytes, plasma cells, eosinophils and, occasionally, neutrophils, which penetrate the core of the marker with time. The degradation of the collagen plug appears to be associated with infiltration of the marker by fibrovascular tissue and deposition of native collagen; of note is the absence of a significant multinucleate giant cell reaction. These novel breast biopsy site markers do not interfere with the histologic processing of the tissue or with their histopathologic interpretation. PMID- 15897750 TI - Peripheral bile duct paucity and cholestasis in the liver of a patient with Alagille syndrome: further evidence supporting a lack of postnatal bile duct branching and elongation. AB - Alagille syndrome (AGS) is a developmental, multiorgan disease caused by mutations of the Jagged1 gene. The liver is one of the major organs affected in AGS, and the hallmark of liver pathology in AGS is an age-related increase in the proportion of portal tracts that have no bile duct, but without evidence of prominent bile duct damage. The pathogenesis of this bile duct paucity is currently not well understood. (Immuno)histochemical and molecular analyses were performed on several liver biopsies that were taken during macroscopic examination of the explant liver of a 17-year-old AGS patient. The liver periphery was macroscopically pale and was microscopically characterized by complete absence of bile ducts and presence of severe cholestasis, but there was no ductular reaction. Conversely, the central, hilar portion contained normally developed bile ducts showing no or minimal damage and cholestasis. A missense mutation in the Jagged1 gene was present in both parts of the liver, indicating that mosaicism did not cause this peculiar picture. There was also a hypertrophy of the hepatic arterial branches in the liver periphery. Together with previous indirect findings, the current study of the explant liver of an AGS patient strongly suggests that a lack of branching and elongation of bile ducts during postnatal liver growth is the mechanism by which peripheral bile duct paucity and cholestasis develops in AGS. Our findings also suggest that anomalies of the intrahepatic arterial branches may be part of AGS in some patients. PMID- 15897751 TI - Mammary presentation of adult-type "juvenile" xanthogranuloma. AB - Juvenile xanthogranuloma (JXG) is a benign histiocytic disorder of infants and childhood. Approximately 15% of cases occur in adults. Adult JXG characteristically affect patients in their 20s and 30s; however, about 5% of patients are older than 60 years. Adult JXGs rarely regress spontaneously, and reports of concomitant extracutaneous lesions are rare. Herein, we report an exceptional case of adult xanthogranuloma in a 74-year-old woman who presented with ipsilateral breast masses and also found to have prior cutaneous lesions. This is the first reported case of cutaneous and extracutaneous adult JXG where the latter manifested in the breast as a spindle cell xanthogranuloma. Histologically, the lesion was composed predominantly of spindle cells with associated multinucleated giant cells and a chronic inflammatory cell infiltrate. Spindle cells were immunoreactive for various histiocytic markers and negative for cytokeratins, S-100, CD34, factor XIIIa, and CD1a. In the breast, the morphologic features of JXG evoked several entities in the differential diagnosis, including spindle cell metaplastic carcinoma, inflammatory pseudotumor, fibromatosis, myofibroblastoma, and phyllodes tumor. With the aid of immunohistochemical stains and appropriate clinical history, the correct diagnosis of extracutaneous adult JXG manifesting as a spindle cell xanthogranuloma can be made. PMID- 15897752 TI - Metallic implant-associated lymphoma: a distinct subgroup of large B-cell lymphoma related to pyothorax-associated lymphoma? AB - Primary non-Hodgkin lymphoma arising at the site of metallic implant is very rare, and the possible carcinogenic effects of the metallic components and wear particles of the implant have not been answered despite many years of investigation. We report a case of large B-cell lymphoma occurring in a 78-year old man who had a knee prosthesis implant for more than 30 years. The lymphoma was of microscopic size and found incidentally in the wear debris removed at surgical revision of the loosened prosthesis. The lymphoma expressed CD20, showed clonal rearrangements of immunoglobulin gene, and harbored Epstein-Barr virus (EBV). This case, together with previously reported cases, suggests that metallic implant-associated lymphoma is a distinctive subgroup of large B-cell lymphoma that shares many similarities with pyothorax-associated lymphoma and osteomyelitis-associated lymphoma, in that the lymphoma is an EBV-associated large B-cell lymphoma arising in a setting of chronic inflammation or irritation in a confined body space. PMID- 15897754 TI - Eosinophilic dysplasia of the cervix: which are the invasive and cytologic counterparts? PMID- 15897755 TI - Sarcomatoid variant of anaplastic large cell lymphoma mimics ALK-1-positive inflammatory myofibroblastic tumor in bladder. PMID- 15897756 TI - Sclerosing angiomatoid nodular transformation (SANT) of the spleen: addition of a case with focal CD68 staining and distinctive CT features. PMID- 15897758 TI - Villous adenocarcinoma of the colon. PMID- 15897759 TI - c-kit overexpression in chromophobe renal cell carcinoma is not associated with c kit mutation of exons 9 and 11. PMID- 15897760 TI - The importance of specialization. PMID- 15897762 TI - Computer simulations: a promising new tool for the CNS. PMID- 15897763 TI - Making the right choice: family presence and the CNS. PMID- 15897764 TI - The Agency for Healthcare Research and Quality: a valuable resource for evidence based practice. PMID- 15897765 TI - Telithromycin--a ketolide antibiotic for the treatment of outpatients with community-acquired respiratory tract infections. PMID- 15897766 TI - Clinical nurse specialist profile. Interview by Jo Ellen Rust. PMID- 15897767 TI - It is all about cash flow. PMID- 15897768 TI - Theoretical approaches for investigating patient safety. AB - Patient safety and freedom from accidental injury is an issue that is promoting the search for excellence among healthcare providers, payers, and consumers. The issue is complex and multifaceted, providing many avenues for analysis, quality enhancement, and research. Several models exist that may assist in exploring patient safety issues and the relationships between error and safety. Three models are discussed and research questions are generated that with further investigation will help us to understand the complexity of error management and the promotion of patient safety. Because of their leadership role and guardianship in managing patient care, clinical nurse specialists understand and promote models that improve safety for their patients. PMID- 15897770 TI - Potassium protocols: in search of evidence. AB - PURPOSE: The purpose of this triangulated study was to evaluate potassium protocols used in clinical practice. RATIONALE: Hypokalemia occurs frequently in the clinical setting and can have serious consequences. It is further complicated by the fact that it has multiple causes and patient symptoms can range from asymptomatic to death. considering the complexity and potential seriousness of hypokalemia, it is important that treatment be appropriate. DESCRIPTION OF THE PROCESS: The triangulated approach included an examination of the empirical evidence, a comparison of potassium protocols currently in use, and an evaluation of the potential benefits and risks of using a potassium protocol in a sample of patients. OUTCOME: There is wide variation in potassium protocols and no empirical evidence in support of or opposition to these protocols. An evaluation of the use of a potassium protocol in a sample of patients indicates that patients not on protocols are not being routinely treated according to general protocol potassium replacement recommendations. CONCLUSIONS: The use of narrowly defined potassium protocols may lead to overtreatment or incorrect treatment in the complex setting of hypokalemia. IMPLICATIONS FOR NURSING PRACTICE: Findings suggest the need for validation of clinically significant hypokalemia, the addition of other electrolyte measures when evaluating and treating hypokalemia, and the use of routine serum potassium levels and maintenance of serum potassium levels equal to or more than 4.0 mmol/L in certain patient populations. Most important, the etiology of hypokalemia, not used in qualifying criteria on any of the potassium protocols examined, should form the basis for treatment. PMID- 15897771 TI - FAMISHED for support: recovering elders after cardiac events. AB - BACKGROUND: Heart disease causes disproportionately heavy burdens on unpartnered elders (widowed, divorced, never married) who have limited social resources to contend with recovery demands and complications resulting from acute myocardial infarction (AMI) or coronary artery bypass graft (CABG) surgery. Research suggests that the availability of social support improves recovery after AMI and CABG, yet functional support expressly designed to modify health and stimulate recovery remains unspecified. Advanced practice nurses (APNs) in the clinical nurse specialist (CNS) role who provide social support to recovering elders may be able to improve health outcomes for this vulnerable group. AIMS: The investigators' aims are to (1) quantify the duration and frequency of nurse contact with unpartnered elders recovering from cardiac events and (2) describe the functional support provided by the CNS to the unpartnered recovering elder. METHODS: Forty narrative accounts comprising 670 entries by interventional CNS APNs were examined using thematic analytic techniques. RESULTS: Over a 14-week period, CNS APNs contacted recovering elders 16 times, with a total of 4.5 hours spent per elder, or 13 minutes per phone call and 80 minutes per home visit. Analysis of 670 entries demonstrated functional and emotional support that was captured in the acronym, FAMISHED. CONCLUSIONS: Functional and emotional support from the CNS may enhance self-efficacy for recovery from cardiac events for unpartnered elders. PMID- 15897772 TI - Protecting the kidneys of patients with diabetes. AB - Diabetes is the most common cause of end-stage renal disease; however, there is evidence that diabetes-related renal disease can be prevented with interventions currently available. The purposes of this article are to describe current screening and intervention guidelines for renal complications in patients with diabetes and to provide clinical nurse specialists with tools to facilitate the education and expert guidance needed by patients with diabetes to preserve their kidney function. This article includes (1) a review of the pathology of diabetic nephropathy, (2) a summary of screening guidelines, (3) current treatment recommendations to prevent or delay nephropathy, and (3) nursing strategies and tools framed within the education, expert coaching, and guidance roles of the clinical nurse specialist practice model. PMID- 15897773 TI - The Healthcare Consumer's Environmental Bill of Rights: considering the implications. PMID- 15897774 TI - Segmental heterogeneity in the mechanism of sodium nitroprusside-induced relaxation in ovine pulmonary artery. AB - Segmental heterogeneity in relaxation response to nitric oxide (NO) was examined using NO donor sodium nitroprusside (SNP) in second- (medium) and fourth generation (small) ovine isolated intralobar pulmonary arteries. In vessels precontracted with serotonin, NO donors SNP and S-nitroso-N-acetylpenicillamine (SNAP) were more potent in relaxing medium, in comparison to the small, arteries. Soluble guanylyl cyclase (sGC) inhibitor [1,2,4]oxadiazolo-[4,3-a]quinoxaline-1 one (ODQ 3 microM) caused a profound inhibition of SNP relaxation in small as compared with medium-sized arteries. However, both basal and SNP (10 microM) stimulated intracellular cyclic guanosine monophosphate (cGMP) content was identical in these 2 arterial segments. The Na,K-ATPase inhibitor ouabain (1 microM) had a marked inhibitory effect on SNP-mediated relaxation in both segments. There was no segmental difference in SNP (10 microM)-stimulated plasma membrane Na,K-ATPase activity and ouabain-sensitive Rb-uptake. 4-AP (1 mM), a relatively selective inhibitor of Kv channels, decreased the potency of SNP relaxation by about 10-fold in the medium-sized vessels. On the other hand, 4-AP was without effect on the vasodilator potency of SNP in small vessels. Interestingly, in the presence of 4-AP, SNP was equipotent in dilating both medium (pD2 = 5.80 +/- 0.07; Emax = 84 +/- 1.6%, n = 7) and small (pD2 = 5.74 +/- 0.15; Emax = 83 +/- 2.5%, n = 7) pulmonary arteries. In conclusion, the results of the present study suggest that Kv channels determine the segmental heterogeneity of NO-mediated relaxation in ovine pulmonary artery. PMID- 15897775 TI - Decrease of endothelin receptor subtype ETB and release of COX-derived products contribute to endothelial dysfunction of porcine epicardial coronary arteries in left ventricular hypertrophy. AB - Alterations in the regulation of coronary circulation play a major role in the enhanced susceptibility to ischemic injury of the myocardium in left ventricular hypertrophy (LVH). The present study was designed to assess the role of endothelium-dependent contracting factors and endothelin receptors in the coronary endothelial dysfunction in LVH, occurring 2 months after aortic banding in a swine model. Hemodynamic and morphologic analyses were performed in LVH and control groups. Vascular reactivity studies were performed in rings from control and aortic banding groups to assess the contribution of endothelin (ET-1) receptor subtypes to the contraction induced by ET-1 and IRL-1620 (an ETB receptor agonist), with and without endothelium. The effects of cyclooxygenase (COX)-derived products induced by ET-1, serotonin (5-HT), and bradykinin (BK) were evaluated, with or without indomethacin (a COX antagonist). ET-1 receptor density was assessed by confocal microscopy and Western blot experiments. The wall-to-lumen ratio, determined in digital planimetry, was increased in the LVH group with no significant changes in coronary perfusion pressures. There was a significant increase in contractions to ET-1 in the LVH group, which were reduced by exposure to indomethacin and daltroban (thromboxane A2 [TXA2] receptor antagonist). Relaxations to 5-HT and BK were improved by indomethacin in the LVH group. There was no significant change in ETA receptor density (3.113 +/- 0.389 vs 3.594 +/- 0.314) but a decrease in ETB receptor density (6.435 +/- 0.265 vs 4.588 +/- 0.089; P < 0.001) in the LVH group. The coronary endothelial dysfunction of swine epicardial coronary arteries in LVH secondary to 2 months of aortic banding involves both relaxing and contracting factors. ETA receptors and COX-derived products are preferentially implicated in the increased contractions to ET-1. Strategies aimed at decreasing ET-1 effects with ET-1 antagonists selective for ETA receptors could improve the coronary endothelial dysfunction in LVH. PMID- 15897776 TI - N-acetyl-L-cysteine mitigates aortic tone injury following liver ischemia reperfusion. AB - Liver ischemia and subsequent reperfusion (IR) are associated with secondary, remote organ reperfusion injury attributable to oxidative stress mediators. Because N-acetyl-L-cysteine (NAC) was effective in attenuating lung reperfusion injury, its properties on aortic dysfunction were tested. Rat isolated perfused aortic rings (n = 8/group) were evaluated during and after exposure to liver postischemia perfusate. Aortic response to phenylephrine under these conditions was also assessed in the presence or absence of increasing concentrations of NAC. Aortic rings incubated with postischemia perfusates exhibited abnormally protracted contraction. Their response to phenylephrine was reduced to 18 +/- 7% and 65 +/- 11% of controls during and after the exposure, respectively, and their subsequent relaxation was irregular. NAC 0.25 mM best attenuated the IR-induced aortic tone impairments, 0.12 mM affected it slightly, and IR-NAC 0.5 mM and IR NAC 0.74 mM solutions dilated the rings proportionately, abolishing reactions to both IR solutions and phenylephrine. Xanthine oxidase activity and reduced glutathione (GSH) level in all IR ring tissues were inversely proportionate, but not directly so. Thus, liver IR impaired aortic tone and its response to phenylephrine, even after removal of toxic elements. NAC concentrations directly and inversely correlated with xanthine oxidase activity but not with GSH level. It preserved aortic functions dose-specifically, mainly by oxidant quenching. PMID- 15897777 TI - Aqueous extract of Salvia miltiorrhoza regulates adhesion molecule expression of tumor necrosis factor alpha-induced endothelial cells by blocking activation of nuclear factor kappaB. AB - Salvia miltiorrhoza Bunge is a traditional herb medicine often used in China for the treatment of cardiovascular disorders, such as atherosclerosis. The purpose of this study was to examine the effect of aqueous extract of Salvia miltiorrhoza Bunge (ESM) on expression of adhesion molecules in tumor necrosis factor (TNF)alpha-induced endothelial cells. When preincubated with ESM (100, 200, 400 microg/mL) for 18 hours, the adhesion of HL-60 cells to TNFalpha-induced endothelial cells was significantly decreased in a concentration-dependent manner, and down-regulation of adhesion molecules, intracellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), was also observed. The inhibitory effect of ESM on TNFalpha-induced VCAM-1 expression was attenuated by inhibition of intracellular glutathione (GSH) synthesis. In addition, ESM also significantly inhibited TNFalpha-induced translocation of nuclear factor kappaB (NF-kappaB) from cytoplasm to nuclei in endothelial cells. These results demonstrated that inhibition of ESM on the expression of adhesion molecules may result from its blocking activation on NF-kappaB. It may imply one of the mechanisms by which ESM exerts its beneficial effect preventing the progress of atherosclerosis. PMID- 15897778 TI - Pravastatin restores DDAH activity and endothelium-dependent relaxation of rat aorta after exposure to glycated protein. AB - This study was designed to investigate whether glycated bovine serum albumin (AGE BSA) inhibits dimethylarginine dimethylaminohydrolase (DDAH) activity to contribute to its adverse effect on endothelium-dependent relaxation in rat aorta, and whether pravastatin reverses the inhibition of DDAH activity and endothelial dysfunction induced by AGE-BSA. Endothelium-dependent relaxation of aortic rings was measured by isometric tension recording, and DDAH activity, and the contents of nitrite/nitrate as well as malondialdehyde (MDA) in aortic tissue were determined after exposure of Sprague-Dawley rat aorta to AGE-BSA (1.70 mmol/L) for 60 minutes in the presence or absence of pravastatin. In comparison with control, both endothelium-dependent relaxation and DDAH activity (0.032 +/- 0.002 versus 0.095 +/- 0.003 U/g protein, n = 5, P < 0.01) were significantly inhibited in isolated rat aorta after exposure to AGE-BSA, which was accompanied by decreases of nitrite/nitrate contents and elevations of MDA levels in aorta. Treatment with pravastatin (1 mmol/L) not only prevented the inhibition of endothelial function but also reversed the decrease of DDAH activity induced by AGE-BSA and normalized the alterations in nitrite/nitrate and MDA contents. Similar effects were observed when rat aorta exposed to AGE-BSA in the presence of antioxidant pyrrolidine dithiocharbamate (PDTC, 30 micromol/L) or protein kinase C inhibitor chelerythrine (1 micromol/L). These results suggested that decreased DDAH activity may be involved in endothelial dysfunction of rat aorta induced by AGE-BSA, and that pravastatin restores DDAH activity and endothelium dependent relaxation after aorta exposure to AGE-BSA, which may be secondary to its antioxidative effects. PMID- 15897779 TI - Negative metabolic effects of cGMP are enhanced in obese rat hearts. AB - Leptin resistance leads to obesity and may affect responses to the second messenger cGMP. We tested the hypothesis that the myocardial negative metabolic response to cGMP would be enhanced in leptin-resistant animals. This hypothesis was tested in anesthetized open-chest Zucker obese (n = 16) and age-matched control rats (n = 13). Coronary blood flow (microspheres) and O2 extraction (microspectrophotometry) measurements were used to determine myocardial O2 consumption (VO2). Protein phosphorylation by cGMP protein kinase and cAMP phosphodiesterase activity were also determined. Either vehicle (saline) or 8-Br cGMP (10(-3) M) was topically applied to the left ventricular surface. Body weight was significantly greater in the obese rats (523 +/- 17 versus 322 +/- 12 g). There were no hemodynamic differences between groups. There was no difference in VO2 between lean (52 +/- 13 mL O2/min/100 g) and obese (54 +/- 9) vehicle treated rats. 8-Br-cGMP significantly lowered VO2 in obese (35 +/- 6) but not lean (45 +/- 7) rats. This was not related to altered protein phosphorylation by the cGMP protein kinase. Cyclic GMP inhibited cAMP phosphodiesterase activity in lean but not obese hearts. Thus, the high myocardial oxygen consumption of lean rats was not significantly affected by cGMP but was reduced in obese hearts. This appeared to be related to a reduced inhibition of cAMP phosphodiesterase activity by cGMP in the Zucker obese rat. PMID- 15897780 TI - Paradoxical effects of iron chelation on growth of vascular endothelial cells. AB - Endothelial cell (EC) and vascular smooth muscle cell (VSMC) interactions play critical roles in restenosis following vascular injury. We examined the effects of intracellular iron chelation on endothelial cell cycle progression and VSMC modulation of endothelial cell growth. A diffusible, lipid-soluble iron chelator that rapidly enters cells, desferri-exochelin 772SM (D-Exo), was studied in human endothelial cells and VSMCs. In both cell types D-Exo reversibly halted cell cycle progression from G0/G1 phase to S phase and from S phase to G2/M phase and increased expression of hypoxia-inducible factor 1alpha (HIF-1alpha). D-Exo increased secretion of vascular endothelial growth factor (VEGF), a downstream target of HIF-1alpha, in VSMCs, but there was no VEGF production in endothelial cells. D-Exo was 25-fold more potent than the lipid-insoluble iron chelator deferoxamine, which does not readily enter cells. Intracellular iron chelation with D-Exo directly inhibits endothelial cell growth but indirectly stimulates endothelial cell growth by increasing VEGF release by VSMCs. PMID- 15897781 TI - Repolarization dynamics in patients with idiopathic ventricular fibrillation: pharmacological therapy with bepridil and disopyramide. AB - The electrocardiographic parameters relating occurrence of ventricular fibrillation (VF) episodes in patients with idiopathic VF (IVF) are still unknown. The aim of this study was to clarify efficacy of pharmacological therapy in patients with IVF with respect to repolarization dynamics. The study group consisted of 8 men (age 43.6 +/- 9.1 years) with IVF (Brugada type 5 patients, prominent J wave in the inferior leads 3 patients) who had documented spontaneous episodes of VF, 7 of whom had implantable cardioverter defibrillators. The relation between QT and RR interval was analyzed from 24-hour Holter ECG using an automatic analyzing system before and after pharmacological therapy (bepridil 5 and disopyramide 3). From QT-RR linear regression lines, QT intervals were determined at RR intervals of 0.6 second [QT(0.6)], 1.0 second [QT(1.0)], and 1.2 seconds [QT(1.2)]. Pharmacological therapy increased the slope of QT-RR regression line from 0.105 +/- 0.020 to 0.144 +/- 0.037 (P < 0.05). Accordingly, QT(1.0) and QT(1.2) became longer after drug therapy [QT(1.0), 0.382 +/- 0.016 seconds vs 0.414 +/- 0.016 seconds (P < 0.01); QT(1.2), 0.403 +/- 0.017 seconds vs 0.442 +/- 0.021 seconds (P < 0.01)]. However, QT(0.6) did not change after drug administration. Before drug therapy the average episodes of VF were 5.5 +/- 5.8 (range 1 to 17) during the observation period of 19.3 +/- 17.6 months (range 6 to 60 months). After drug therapy, 6 patients had no episode of VF for 24 to 120 months (66.0 +/- 38.5 months). Two patients had a single episode of VF for 12 and 96-month follow-ups. Pharmacological therapy decreased the frequency of VF episodes in association with prolongation of QT intervals at slower heart rates. Not only J wave and ST elevation but also shorter QT intervals at slower heart rates may represent an electrophysiological substrate for development of VF episodes in these specific IVF patients. PMID- 15897782 TI - Comparison of the vasodilatory effect of nadroparin, enoxaparin, dalteparin, and unfractioned heparin in human internal mammary artery. AB - The aim of this study was to investigate whether the low-molecular-weight heparins (LMWHs) (eg, nadroparin, enoxaparin, and dalteparin) cause a vasodilatory effect in human internal mammary artery (IMA) and to further compare its effect with unfractioned heparin (UFH). Samples of redundant IMA obtained from 20 patients undergoing a coronary artery bypass graft surgery were cut into 3-mm-wide rings and suspended in 20-mL organ baths. Isometric tension was continuously measured with an isometric force transducer connected to a computer based data acquisition system. LMWHs (0.5-6 U/mL) caused a concentration dependent relaxation in the endothelium-intact human IMA rings, which were precontracted with Phe (10(-6) M) (P < 0.05). The vasodilator potency of LMWHs seems to be nearly similar while the maximal effect produced by LMWHs was less pronounced compared with that produced by UFH. Removal of endothelium totally abolished the responses of human IMA to LMWHs as well as UFH (P < 0.05). LMWHs induced vasodilator effect was significantly attenuated by Nomega-nitro-L arginine methyl ester (L-NAME, 10(-4) M) but not indomethacin (10(-5) M). Our results have shown that LMWHs cause a dose-dependent relaxation in human IMA but are less effective than that produced by UFH. The vasorelaxant effects induced by each of LMWH are nearly similar and seem to be via endothelium-dependent mechanisms, including generation of nitric oxide. PMID- 15897783 TI - Oral administration of geranylgeranylacetone blunts the endothelial dysfunction induced by ischemia and reperfusion in the rat heart. AB - It has been shown that geranylgeranylacetone (GGA) protects heart against ischemia/reperfusion injury via enhanced heat shock protein 72 (HSP72) expression in rats. In the present study, we investigated the protective effect of GGA on ischemia/reperfusion-induced endothelial dysfunction. Rats were given oral GGA (GGA group) or vehicle (CON group), and 24 hours later their hearts were removed and placed in the Langendorff apparatus for 30-minute low-flow ischemia followed by 30-minute reperfusion. GGA improved the postischemic functional recovery (P < 0.01), which was abolished by N-nitro-L-arginine methyl ester (L-NAME, NO synthase inhibitor). NO production during both ischemia and reperfusion were increased in the GGA group, and the acetylcholine (ACh)-induced (endothelium dependent) vasodilation, measured as the percentage decrease in coronary perfusion pressure after ischemia/reperfusion (14.9 +/- 1.3%), was preserved as compared with that in the CON group (7.9 +/- 1.4%). LY294002, a phosphatidylinositol 3 (PI3) kinase inhibitor, abolished the protective effects of GGA on endothelial-dependent coronary vasodilation and NO production, whereas Y27632 (Rho kinase inhibitor) increased endothelium-dependent coronary vasodilation and NO production in CON group toward the level seen in GGA group. The amount of adrenomedullin in the coronary effluent at basal condition was lower in the GGA group than in the CON group (P < 0.05), and during both ischemia and reperfusion there was no difference in the amount of adrenomedullin between the GGA and CON groups. In addition, no difference was observed in the amount of endothelin-1 between the GGA and CON groups. These results indicate that GGA attenuates the ischemia/reperfusion-induced coronary endothelial dysfunction, which may contribute to its cardioprotective effect. The PI3 kinase and/or Rho kinase pathways appear to be involved in this process, whereas adrenomedullin and endothelin-1 are not necessary for the GGA-induced cardioprotection. PMID- 15897784 TI - Left ventricular diastolic function improvement by carvedilol therapy in advanced heart failure. AB - Carvedilol treatment in chronic heart failure (CHF) patients has been demonstrated to reduce mortality by improving cardiac systolic function and reducing left ventricular adverse remodeling. However, the effects of the drug on left ventricular (LV) filling are less studied. In this study we evaluated early and long-term diastolic cardiac modifications by an echo-Doppler method during carvedilol therapy in patients with advanced CHF and pseudonormal or restrictive filling pattern. We studied 58 patients with severe but stable CHF (39 in class NYHA III and 19 in IV) having systolic and diastolic dysfunction caused by idiopathic or ischemic cardiomyopathy. Thirty-two patients were randomized to receive previous treatment plus carvedilol (group 1) and 26 continued standard therapy (group 2). In all subjects we evaluated LV volumes, LV mass, LV ejection fraction (EF), and the following transmitral filling parameters: early wave (E), atrial wave (A), E/A ratio, deceleration time (DT), and isovolumetric releasing time (IVRT). After 4 months of therapy, the carvedilol group showed a significant increase of A wave (P < 0.001), DT (P < 0.0001), IVRT (P < 0.0001), and a significant reduction of E/A ratio (P < 0.0005) with respect to group 2. Further improvement was observed at 12 months (A P < 0.0005; DT P < 0.00002; IVRT P < 0.000004; E/A P < 0.0008), although an E wave reduction was observed in group 1 with respect to controls (P < 0.001). Moreover, after 1 year of follow-up a reduction of systolic volume (P < 0.001) and pulmonary pressure (P < 0.0001) and consequent increase of EF (P < 0.001) was observed in the carvedilol group. Carvedilol treatment improved diastolic function in CHF with severe diastolic and systolic impairment at early time, converting a restrictive or pseudonormal filling pattern into an altered pattern. These changes remained significant after 1 year of therapy together with improvement in systolic function. PMID- 15897785 TI - Impact of nitric oxide synthase inhibitor and chloride channel antagonist on mesenteric vascular conductance in anesthetized Dahl normotensive and hypertensive rats. AB - The effects of nitric oxide synthase inhibitor N-nitro-L-arginine methyl ester (L NAME) and chloride channel antagonist niflumic acid on vascular responsiveness to the effect of alpha1-adrenoceptor stimulation in the mesenteric bed of Dahl salt resistant normotensive (SRN) and salt-sensitive hypertensive (SSH) rats were examined. Dahl salt-resistant and salt-sensitive rats were fed a high-salt diet (4% NaCl) for 7 weeks, and blood pressure, heart rate, and mesenteric blood flow were measured before and after treatment with L-NAME (0.3 mg/kg, IV) and/or niflumic acid (10 mg/kg, IV). Morphometry of the primary mesenteric blood vessel was also assessed. Administration of alpha1-adrenoceptor agonist cirazoline produced a dose-dependent increase in blood pressure, decrease in heart rate, mesenteric blood flow, and mesenteric vascular conductance in SRN and SSH rats. L NAME significantly increased basal blood pressure and decreased basal mesenteric blood flow and vascular conductance in SRN but not in SSH rats. Niflumic acid attenuation of cirazoline-mediated decreases in mesenteric blood flow and vascular conductance was more pronounced in the SRN than SSH rats. This difference in the inhibitory actions of niflumic acid was absent following its concomitant administration with L-NAME. It seems that tonic release of nitric oxide modulates niflumic acid-sensitive chloride channels in vascular muscle. Blood vessels from SSH rats had significantly larger smooth muscle thickness and lumen diameter, but the ratio of the 2 were not different between the SRN and SSH. Our findings support the view that alterations in receptor-mediated signal transduction, rather than just changes in blood vessel architecture, are responsible for differences in behavior of blood vessels in salt-induced hypertensive rats. PMID- 15897786 TI - Testosterone relaxes human internal mammary artery in vitro. AB - Preliminary clinical studies of testosterone therapy in male patients with coronary artery disease raised promising results. However, there is no study on in vitro effects of testosterone in human isolated arteries. We investigated the effect of testosterone on contractile tone of human isolated internal mammary artery. The responses in human internal mammary artery (IMA) were recorded isometrically by a force-displacement transducer in isolated organ baths. Testosterone (10 nM to 100 microM) was added cumulatively to organ baths either at rest or after precontraction with KCl (68 mM) and PGF2alpha (10 microM). Testosterone-induced relaxations were tested in the presence of cyclooxygenase inhibitor indomethacin (10 microM), nitric oxide synthase inhibitor Nomega-nitro L-arginine methyl ester (L-NAME, 1 microM), nonselective large-conductance Ca2+ activated and voltage-sensitive K+ channel inhibitor tetraethylammonium (TEA, 1 mM), ATP-sensitive K+ channel inhibitor glibenclamide (GLI, 100 microM), and voltage-sensitive K+ channel inhibitor 4-aminopyridine (4-AP, 1 mM). Testosterone produced relaxation in human IMA (Emax 33% and 41% of KCl- and PGF2alpha-induced contraction, respectively). Vehicle had no significant relaxant effect. Except for TEA, the relaxation at low concentrations is not affected by either K+ channel inhibitors (GLI and 4-AP) or L-NAME and indomethacin. We report for the first time that supraphysiological concentrations of testosterone induce relaxation in IMA. This response may occur in part via large-conductance Ca2+ activated K+ channel-opening action. PMID- 15897787 TI - Vaccinating rabbits with a cholesteryl ester transfer protein (CETP) B-Cell epitope carried by heat shock protein-65 (HSP65) for inducing anti-CETP antibodies and reducing aortic lesions in vivo. AB - Rabbits were vaccinated with a recombinant protein vaccine of HSP65-CETPC comprising mycobacterial heat shock protein-65 (HSP65) and a cholesteryl ester transfer protein (CETP) B-cell epitope in alum adjuvant for inducing anti-CETP antibodies in vivo. After anti-CETP antibodies were successfully produced, rabbits were fed with a high-cholesterol diet for 15 weeks, and then the antiatherogenic effects of anti-CETP antibodies were evaluated. The results showed that the fraction of plasma cholesterol in HDL increased and the fraction of plasma cholesterol in LDL decreased in rHSP65-CETPC-immunized rabbits when compared with those in control animals. The average percentage of aortic lesions in the entire aorta area in rHSP65-CETPC-vaccinated rabbits was 23.8% less than in OVA-immunized rabbits (15.14% vs 19.86%) and 30.8% less than in rHSP65 immunized rabbits (15.14% vs 21.87%). The average thickness of hyperplastic coronary artery in rHSP65-CETPC immunized rabbits was 164 +/- 12 microm, significantly lower than in rHSP65-immunized rabbits (197 +/- 15 microm) and in OVA-immunized rabbits (206 +/- 15 microm). Taken together, vaccination with the rHSP65-CETPC vaccine could significantly attenuate atherosclerosis in a rabbit model. Thus, the chimeric protein of rHSP65-CETPC can be further developed into an antiatherosclerosis vaccine in the future. PMID- 15897788 TI - Effects of doxazosin and irbesartan on blood pressure and metabolic control in patients with type 2 diabetes and hypertension. AB - The objective of this trial was to compare the metabolic effects of long-term treatment with doxazosin to those of irbesartan in patients with type 2 diabetes and hypertension. We evaluated 96 hypertensive diabetic patients who were randomized to 12 months of double-blind treatment with doxazosin 4 mg/d or irbesartan 300 mg/d. At the end of the study, systolic and diastolic blood pressure (SBP and DBP) were significantly reduced from 152 to 140 mm Hg and from 97 to 87 mm Hg, respectively, with doxazosin (P < 0.01). SBP and DBP were reduced from 150 to 134 mm Hg and from 94 to 83 mm Hg, respectively, with irbesartan (P < 0.01). Irbesartan had significantly better antihypertensive efficacy than doxazosin (P < 0.05). Doxazosin had the greatest effect on glucose metabolism and lipid parameters, with significant (P or =2-min epochs of fetal oxygen saturation <30% (immediate: M = 7.9; delayed: M = 2.7), F(1, 43) = 6.23, p = .02. There were more variable decelerations of the fetal heart rate in the immediate pushing group (immediate: M = 22.4; delayed: M = 15.6) F(1, 43) = 5.92, p = .02. There were no differences in length of labor, method of birth, Apgar scores, or umbilical cord blood gases. Women who pushed immediately had more perineal lacerations (immediate: n = 13; delayed: n = 5) chi(1, N = 45) = 6.54, p = .01. DISCUSSION: Delayed pushing results in less fetal oxygen desaturation and less > or =2-min epochs of fetal oxygen saturation <30% during second-stage labor than the immediate pushing method; thus, delayed pushing is more favorable for fetal well-being as measured by fetal oxygen saturation. PMID- 15897791 TI - Family education and support interventions in heart failure: a pilot study. AB - BACKGROUND: Self-management of dietary sodium restriction by persons with heart failure (HF) is difficult and usually occurs within the home setting and within a family context. OBJECTIVE: To compare a patient and family education (EDUC) intervention with a combined education and family partnership intervention (EDUC + FPI) for effects on improving dietary sodium self-management in persons with HF. METHODS: Patients with HF and a family member (FM) were randomized to EDUC (n = 29 dyads) or EDUC + FPI (n = 32 dyads). Participants with HF were primarily White males with a mean age of 61 years (+/-12). The FMs were primarily women and spouses and had a mean age of 54 years (+/-17). Self-reported dietary sodium (Diet NA) intake and 24-hr urinary sodium (Urine NA) were measured at baseline (BL) and 3 months (3M) after intervention. Data were analyzed with descriptive statistics, generalized least squares regression, paired t test, and chi-square test. RESULTS: Groups did not differ by age, gender, or clinical variables; however, family functioning (Family APGAR) scores were slightly higher in the EDUC + FPI group at BL. Both groups decreased Diet NA and Urine NA from BL to 3M; the EDUC + FPI group showed greater decrease in Urine NA and had a greater percentage of those who decreased Urine NA by at least 15% (p = .04). Regression analysis to predict Urine NA revealed a significant Group x Time interaction (p = .03) when accounting for time-varying measures of body mass index (p = .001). DISCUSSION: A family-focused intervention may be useful in reducing dietary sodium intake in persons with HF. The Urine NA results support the importance of incorporating family-focused education and support interventions into HF care. PMID- 15897792 TI - Profiles of self-rated health in midlife adults with chronic illnesses. AB - BACKGROUND: Self-rated health (SRH), an important indicator of cognitive appraisal of health, consistently predicts mortality, morbidity, and health services utilization. However, few explanations account for how these cognitive appraisals of health might differ within a population of midlife adults with chronic illnesses who may be at risk for further illnesses over time. OBJECTIVES: The purpose of this study was two-fold: (a) to uncover classes of chronically ill midlife adults who shared unique profiles of characteristics that predicted SRH over time and (b) to reveal the predictive factors of SRH for each class over time. METHODS: Using 5 waves of data (1992-2000) from the Health and Retirement Study, the sample included 6,335 respondents (ages 51 to 61 at baseline) who reported at least one chronic illness. Selected components of the Interaction Model of Client Health Behavior guided the inclusion of relevant predictors of SRH from the literature. Latent class regression was employed to simultaneously classify respondents and identify factors that predicted SRH for each class over time. RESULTS: The final model reflected 3 distinct profiles of SRH over time: positive health, average health, and negative health. Four time-varying predictors differed significantly across the 3 classes: overweight, work limitation, depressed mood, and living with a partner. Three time-varying predictors--comorbidity, vigorous activity less than 3 times per week, and current smoking--had the same influence on all 3 classes. DISCUSSION: The differential effects of these predictors on SRH over time distinguish these results from prior research. In future studies, profiles of SRH that are unique to each class could be used to develop class-specific targeted interventions to improve cognitive appraisal of health, whereas generic interventions would be based on the class-independent predictors of SRH. PMID- 15897793 TI - Does it pay to pay? A randomized trial of prepaid financial incentives and lottery incentives in surveys of nonphysician healthcare professionals. AB - BACKGROUND: Monetary incentives in survey research may provide important gains from a methodological perspective in the control and reduction of survey error associated with potential nonresponse of participants. However, few studies have systematically investigated the use of monetary incentives or other methods to improve the response rates in the nonphysician clinician population. OBJECTIVE: To investigate differences in response rates to a mailed self-administered survey of nonphysician clinicians who were randomized to receive a prepaid monetary incentive, a postsurvey prize drawing, or no incentive. METHODS: A randomized controlled trial of financial incentives was conducted from November 2002 to February 2003. Nonphysician clinicians (nurse practitioners [NPs] and physician assistants [PAs]; N = 3,900) randomly selected to participate in a national ethics-related study were assigned randomly in equal allocations (n = 1,300 [650 NPs, 650 PAs]) to three incentive groups: (a) no incentive; (b) a $5 prepaid token incentive in the initial mailing; or (c) a chance to win one of ten $100 prize drawings upon completion and return of a self-administered survey. RESULTS: A $5 cash incentive increased survey response rates to an adjusted 64.2%: a 19.5 percentage point increase over the lottery group (44.7% response rate), and a 22 percentage point increase over the control group (42.2% response rate). DISCUSSION: A nominal cash incentive of $5 yields a significantly higher response rate from nonphysician providers than receiving either a lottery option or no incentive. PMID- 15897794 TI - Anger dysregulation, depressive symptoms, and health in married women and men. AB - BACKGROUND: Anger problems (anger dysregulation) and depressive symptoms have been linked to risk for all causes of mortality, but less is known about the association between anger dysregulation and depressive symptoms within the context of gender differences and health outcomes. OBJECTIVES: The association between anger dysregulation, depressive symptoms, and self-reports of health in married adults was evaluated using an emotion-regulation model. METHODS: Fifty two married couples completed a series of procedures that included an interview assessing their ability to regulate anger, a questionnaire reporting depressive symptoms (Beck Depression Inventory) and self-reports indicating health. RESULTS: Results provided support for hypothesized links between the variables, but they varied by gender: (a) greater anger dysregulation in the wives, but not the husbands, was predictive of depressive symptoms; (b) anger dysregulation was predictive of the husbands' self-reports of health but was not predictive of the wives' self-reports of health; (c) depressive symptoms were not significantly associated with self-reports of health for either married women or men. DISCUSSION: These results suggest that anger dysregulation may play different roles in the depressive symptoms and self-reports of health for married women and men. PMID- 15897795 TI - Developing and testing a self-care intervention for older adults in caregiving roles. AB - BACKGROUND: Theory-based interventions incorporating a self-care perspective for health promotion and protection are needed for older adults. The Self-Care for Health Promotion in Aging (S-CHPA) model provided the basis for development of an intervention focused on promoting health among older adults who provide care for their spouses with dementia. OBJECTIVES: To describe the development of a theory based intervention (Self-Care Talk) and to describe the process of evaluating the implementation of the intervention when it was pilot tested with older spouse caregivers of persons with dementia. METHODS: The S-CHPA model was used to guide development of Self-Care Talk, a series of telephone-based sessions between advanced practice nurses and older adults. Session content included practicing healthy habits, building self-esteem, focusing on the positive, avoiding role overload, communicating, and building meaning. Specific self-care strategies were explored in the context of an individual's experiences, relationships, and condition. Intervention implementation was evaluated using Lichstein's criteria of delivery, receipt, and enactment. RESULTS: Six spouse caregivers of persons with dementia participated in the pilot study. The education and support intervention was delivered as intended (delivery). Participants reported understanding session content (receipt), and planned to use the information (enactment). Participants also reported an increased awareness about self-care practices and offered specific examples of how they intended to practice better self-care. DISCUSSION: The theory-based Self-Care Talk intervention can be implemented as intended and is ready for use in future randomized clinical trials with older spouse caregivers of persons with dementia. PMID- 15897796 TI - Clinical outcomes of aging in place. AB - BACKGROUND: Programs such as Medicaid Home and Community-based Services (HCBS) have provided an alternative to institutionalization through community-based, long-term care services; however, there are limited studies on the clinical outcomes of participants in these programs as compared to nursing home (NH) residents. OBJECTIVE: To compare clinical outcomes of individuals in a community based, long-term care program to individuals of similar case mix in institutional based, long-term care. METHODS: A program called Aging in Place (AIP) was developed by the Sinclair School of Nursing in cooperation with the state of Missouri's HCBS program. The AIP intervention consisted of nurse coordination of the HCBS program and Medicare home health services. A total of 78 AIP participants were matched with 78 NH residents on admission period, activities of daily living (ADLs), cognitive status, and age. The Minimum Data Set (MDS) was collected on the AIP group at admission and every 6 months over a 30-month period. Cognition was measured by the MDS Cognitive Performance Scale (CPS), ADLs by the sum of 5 MDS ADL items, depression by the MDS-Depression Rating Scale, and incontinence by rating on 2 MDS items related to urinary continence. The Cochran Mantel-Haenszel method was used to test the association between the AIP intervention and clinical outcomes. RESULTS: The AIP group clinical outcomes were better at a statistically significant level (less than .05) for the following outcomes: (a) cognition at 6, 12, and 18 months (p = .00); (b) depression at 6 and 12 months (p = .00); (c) ADL at 6 (p = .02), 12 (p = .04), and 24 (p = .00) months; and (d) incontinence at 24 (p = .02) months. In all 4 outcome measures, the AIP group stabilized or improved outcome scores whereas the NH group's outcome scores deteriorated. DISCUSSION: Study results suggest that community based care with nurse coordination enhances clinical outcomes of long-term care participants. PMID- 15897797 TI - The impact of centering first-level predictors on individual and contextual effects in multilevel data analysis. AB - BACKGROUND: Multilevel data analysis is a powerful analytical tool. Properly applying the models and correctly interpreting the findings are two interrelated general issues in using multilevel modeling (MLM). There are two specific issues when using MLM: (a) separating the individual-level effects of a predictor variable from its contextual effects and (b) centering first-level predictor variables. This can have major implications for interpreting the results at higher levels, and its impact on second-level interpretation is not always apparent. OBJECTIVES: The major purposes of this article are to show how to separate organizational-level effects from individual-level effects and to show how first-level centering decisions affect the interpretation of second-level coefficients. METHODS: The hierarchical linear models (HLM) are used to analyze a hypothetical data set with 385 patients nested within 10 hospitals, using uncentered, group-mean-centered, and grand-mean-centered versions of the predictor variable. RESULTS: Uncentered and grand-mean-centered models are equivalent, but group-mean-centered models are not equivalent to the other two. For the grand-mean-centered and uncentered models, second-level coefficients provide correct estimates of the individual effect and the contextual effect when the contextual predictor variable is included in the second-level model. The group-mean-centered model leads to a second-level coefficient where individual level effects are confounded with contextual-level effects. DISCUSSION: There is no single best answer to the question of whether to use group-mean centering or grand-mean centering. The theory and specific questions to be answered should be the researcher's guide to selecting which centering approach to use. Understanding the implications of first-level centering is essential to interpreting second-level coefficients correctly. PMID- 15897798 TI - Mortality related to gender, age, sepsis, and ethnicity in severely burned children. AB - Several studies have noted gender differences in adult mortality related to thermal injury, however, little is published on gender-related outcomes of burn patients 17 years of age or less. The aim of this study was to evaluate the relationships between mortality, gender, prepubertal and during puberty, ethnic origin, and age, with or without identified sepsis in severely burned children. Seven hundred forty-seven children admitted to our burn hospital from March 1985 to January 2005 with burns greater than 40% total body surface area were studied. Mortality associated with identified sepsis, gender, age, and ethnic origin were outcomes of interest. Two hundred sixty (35%) of the patients studied were girls and 487 (65%) were boys. No significant difference could be shown between girls and boys for the number of operations, time from burn to hospital admission, or the presence of identifiable inhalation injury. Nearly 60% of the male nonsurvivors and 48% of the female nonsurvivors in this study had identifiable sepsis at postmortem. The mortality rate was higher in infants and toddlers, age 0 to 2.9 years, compared with children and adolescents, age 3 to 17 years; however, there was no significant difference in rate of mortality between genders, prepuberty versus puberty, those with septic episodes, or ethnic origin. Burn mortality among infants and toddlers, children, and adolescents with greater than 40% total body surface area burns with or without identified sepsis could not be shown to be gender or ethnic origin dependent. PMID- 15897799 TI - Plasma cytokine measurements augment prognostic scores as indicators of outcome in patients with severe sepsis. AB - Despite recent advances in the prospective identification of the patient with sepsis who may benefit from anti-inflammatory or antithrombotic therapies, successful treatment regimens have been fairly modest. We have explored whether determination of several proinflammatory cytokine or mediator concentrations can complement physiologic scoring systems to identify patients with severe sepsis who will survive or expire within 28 days. The design of the study included an exploratory analysis performed in conjunction with a prospective, randomized, double-blind, placebo-controlled, multicenter, clinical trial and involved 33 academic institutions in the United States. One hundred twenty-four patients with severe sepsis with or without septic shock were included in this analysis. Blood samples were obtained at baseline and on days 1 through 4, and were evaluated for proinflammatory and anti-inflammatory cytokine concentrations, as well as for procalcitonin and total protein C levels. Baseline concentrations and changes in the concentrations of these mediators were evaluated in relationship to the Acute Physiology and Chronic Health Evaluation (APACHE) II and multiple organ dysfunction (MOD) scores, and 28-day all-cause mortality. Using univariate logistic regression analyses, APACHE II and MOD scores, age (but not gender), and baseline plasma interleukin (IL)-6 and soluble tumor necrosis factor receptor (sTNFR) 1 (log transformed) concentrations were all predictive of increased 28 day all-cause mortality (P < 0.01). Baseline total protein C, IL-8, IL-10, TNF alpha, and procalcitonin concentrations, and the change in plasma cytokine concentrations from baseline over the initial 4 days were not useful in predicting outcome. Selected baseline proinflammatory cytokine concentrations and APACHE II score were correlated (P < 0.01). IL-6 concentration is a strong candidate for predicting clinical outcome in patients with severe sepsis alone, or when combined with the APACHE II or MOD scores. The potential usefulness of the combination of cytokine measurements and prognostic scores to identify patients who may benefit from treatment with anti-inflammatory or antithrombotic therapies should be further evaluated. PMID- 15897800 TI - Extracorporeal plasma treatment for the removal of endotoxin in patients with sepsis: clinical results of a pilot study. AB - Despite the advances in therapeutic approaches in the management of inflammatory conditions, the incidence of sepsis is on increase in the intensive care units (ICU). In a pilot study, we investigated whether the use of an apheresis system based on DEAE-cellulose is capable of reducing the plasma concentration of endotoxin in patients with severe sepsis. We enrolled 15 intensive care patients with severe sepsis and plasma endotoxin concentrations >0.3 EU/mL. In addition to standard ICU therapy, a total of 83 apheresis treatments were performed. About 1.7 volumes of plasma (6000 mL) were treated at each apheresis session. A significant reduction in plasma endotoxin levels from a median of 0.61 to 0.39 EU/mL (-35%) could be achieved (P < 0.001). Long-term comparison of the initial and post-treatment levels after a series of five to six individual apheresis treatments also showed a statistically significant decline in circulating endotoxin, interleukin (IL)-6, C-reactive protein (CRP), fibrinogen, and an increase in cholesterol levels. Except for a transient and reversible increase of prothrombin time, no adverse events were observed in patients undergoing this new adsorption apheresis treatment. Our data show that reduction of endotoxin by extracorporeal DEAE-cellulose-based plasma treatment may prove a promising therapeutic tool for patients suffering from bacterial sepsis and proven endotoxemia. PMID- 15897801 TI - Interleukin 8 mRNA gene expression in peripheral and intra-abdominal neutrophils during human secondary peritonitis. AB - The aim of the study was to investigate interleukin 8 (IL-8) mRNA gene expression in circulating and emigrated intra-abdominal neutrophils during human secondary peritonitis intra- and postoperatively (until 96 h). Patients with secondary peritonitis were allocated to two groups, e.g., patients with no complications (n = 10) and patients with complications (organ failure, septic shock, etc., n = 9). Patients with elective abdominal surgery (n = 11) and a group with healthy volunteers (n = 7) were studied as controls. Neutrophil RNA was isolated and semiquantitative reverse transcription-PCR was performed. The PCR products were compared with corresponding GAPDH bands (=100%). The highest amount of IL-8 mRNA could be assessed in blood neutrophils of healthy volunteers (87.4% +/- 7.4%). Complicated peritonitis was associated with the lowest concentration of IL-8 mRNA in blood neutrophils intraoperatively (24% +/- 7%, P < 0.05), which showed no recovery throughout the observation period (34% +/- 8%, 96 h postoperatively). IL 8 mRNA concentration in blood neutrophils of patients with uncomplicated peritonitis and patients with elective abdominal surgery was higher intraoperatively (55.2% +/- 9% (uncomplicated peritonitis); 68% +/- 15% (elective abdominal surgery, P < 0.05 versus complicated peritonitis). Thereafter, IL-8 mRNA decreased slightly in both groups, but was distinctly higher than in patients with complicated peritonitis. Emigration to the abdominal cavity resulted in an approximately 2-fold, in some cases 3-fold, increase in the concentration of IL-8 mRNA in emigrated intra-abdominal neutrophils when compared with circulating cells. This increase could be observed in all groups. The long lasting down-regulation of constitutive gene expression of IL-8 mRNA in blood neutrophils during complicated peritonitis is worrying because IL-8 is an important activator and chemoattractant for neutrophils themselves. It is encouraging that migration to another compartment, e.g., infected abdominal cavity, resulted in an increase in neutrophil IL-8 mRNA during complicated and uncomplicated peritonitis. PMID- 15897802 TI - Functional modification of dendritic cells with recombinant adenovirus encoding interleukin 10 for the treatment of sepsis. AB - Control of dendritic cell (DC) function is critical for strategies to modulate innate and acquired immune responses. We examined whether transduction of murine DCs with adenoviral vectors (Adv) expressing interleukin (IL)-10 could alter their phenotype and T cell stimulatory function. Murine bone marrow-derived DCs were transduced with AdV encoding human IL-10 or green fluorescent protein (GFP). Whereas transduction of immature DCs with AdV/GFP resulted in dose-dependent maturation, DCs transduced with Adv/IL-10 maintained an immature state with low major histocompatibility complex (MHC) class II, CD86, and IL-12 expression. The Adv/IL-10 transduced DCs were phenotypically unique, characterized by suppression of IL-12 expression, failure to stimulate Th1 or Th2 cytokine responses, and retained capacity to endocytose antigen. Importantly, Adv/IL-10-transduced DCs were biologically active in vivo, in that administration of these DCs into mice before a generalized peritonitis significantly improved survival. We conclude that Adv/IL-10 transduction of DCs provides an efficient means to modulate DC function. The capacity to modify DCs by adenoviral expression of IL-10 may provide a novel ex vivo or in vivo approach to mitigate acute and chronic inflammatory diseases like sepsis. PMID- 15897803 TI - Fever control in septic shock: beneficial or harmful? AB - The beneficial effects of interventions to control fever in sepsis are controversial. We investigated whether the use of acetaminophen and external cooling is beneficial to control fever in septic shock. We studied 24 fasted, anesthetized, invasively monitored, mechanically ventilated female sheep (27.0 +/ 4.6 kg) that received 0.5 g/kg body weight of feces into the abdominal cavity to induce sepsis. Ringer's lactate (RL) was titrated to maintain pulmonary artery occlusion pressure (PAOP) at baseline levels throughout the experimental period. During the 2 h after the surgical operation, animals were placed in the hypothermia group if their temperature fell below 36.0 degrees C; the other animals were randomized to three groups: high fever (T > 39.0 degrees C), mild fever (37.5 degrees C < T < 38.5 degrees C), and normothermia (36.0 degrees C < T < 37.0 degrees C). The administration of 25 mg/kg acetaminophen every 4 to 6 h combined with external cooling (ice pad) was used to control core temperature in these three groups. The PaO2/FiO2 ratio was higher and blood lactate concentration was lower in the high fever than in the other groups (P < 0.01 and 0.05, respectively). Survival time was longer in the high fever group (25.2 +/- 3.0 h) than in the mild fever (17.7 +/- 3.5 h), normothermia (16.0 +/- 1.9 h), and hypothermia (18.5 +/- 2.5 h) groups (P < 0.05 for all). Plasma heat shock protein (HSP) 70 levels were higher in the two fever groups than in the other groups (P < 0.05). In this clinically relevant septic shock model, the febrile response thus resulted in better respiratory function, lower blood lactate concentration, and prolonged survival time. Antipyretic interventions including acetaminophen and external cooling were associated with lower circulating HSP70 levels. These data challenge the temperature control practices often used routinely in acutely ill patients. PMID- 15897804 TI - Interleukin 10 extends the effectiveness of standard therapy during late sepsis with serum interleukin 6 levels predicting outcome. AB - Patients with septic shock often display features of T cell hyporesponsiveness and immune suppression, which, if persistent, are associated with increased mortality. In the murine cecal ligation and puncture (CLP) model of sepsis, we previously reported that early treatment with the anti-inflammatory cytokine interleukin 10 (IL-10) delays the onset of irreversible shock, defined as the time at which rescue surgery to remove the necrotic cecum is no longer effective. Because IL-10 can be immunostimulatory for T cells, we hypothesized that in the CLP model, late IL-10 treatment after removal of the infectious nidus at the onset of irreversible shock would restore T cell responsiveness and increase survival. C57BL/6J mice were subjected to lethal CLP with and without rescue surgery, concurrent with IL-10 treatment, at the onset of irreversible shock. Survival and serum IL-6 levels were measured as markers of the response to treatment. Ten hours after intervention, all groups exhibited T cell hyporesponsiveness marked by impaired interferon (IFN)-gamma production by Con A stimulated splenocytes. IL-6 levels at 10 h were related to outcome independent of treatment. By 25 h after intervention, only the dual treatment group of cecal removal and IL-10 exhibited T cell responsiveness that was similar to pre-CLP levels (P = 0.26) and had a 7-day survival of 90% (P < or = 0.002 compared with all other groups). Thus, even in the advanced stages of septic shock when standard therapies fail, treatment with IL-10 extends the therapeutic window. For an individual mouse, the efficacy of such treatment may be predicted by an early postintervention IL-6 level. PMID- 15897805 TI - Carbon monoxide prevents multiple organ injury in a model of hemorrhagic shock and resuscitation. AB - The insult from severe hemorrhage is a multifactorial injury involving ischemia/reperfusion with inflammatory dysfunction. Our laboratories and others have demonstrated that the administration of exogenous carbon monoxide (CO) at low concentrations provides cytoprotection in vivo and in vitro. The purpose of these investigations was to test the hypothesis that CO protects against hemorrhagic shock- and resuscitation-induced systemic inflammation and end-organ damage. C57BL/6 mice underwent anesthesia and arterial cannulation. Mice were bled to reach a mean arterial pressure (MAP) of 25 mmHg and were maintained at this pressure for 2.5 h. Mice were then resuscitated with shed blood plus two times the volume of shed blood with Ringer's lactate. Sham animals were not bled. Additionally, mice were maintained in room air or in an environment of CO (250 parts per million). Primary mouse hepatocytes were harvested and used for in vitro cell viability and ATP measurement. These data demonstrate that delivery of a low concentration of inhaled CO protects against the development of end-organ injury decreases serum levels of inflammatory cytokines and increases serum levels of the anti-inflammatory cytokine IL-10. Additionally, CO paradoxically abrogates hemorrhage-induced hepatic cellular hypoxia. Furthermore, CO protected mouse hepatocytes from hypoxia-induced death while maintaining normal ATP levels. CO protects against systemic effects of hemorrhagic shock and resuscitation. The precise cellular mechanisms involved require further elucidation. CO may prove to be an adjunctive therapy that could be instituted rapidly and with ease as an out of-hospital therapeutic modality for severe blood loss after trauma. PMID- 15897806 TI - Release of the mitochondrial enzyme carbamoyl phosphate synthase under septic conditions. AB - To identify sepsis-related dysregulations of protein expression in the liver, we used a baboon model of acute endotoxemia and performed comparative proteome analysis. Treatment with lipopolysaccharide (LPS) was followed by an early but long-lasting (5-48 h) generation of N-terminal fragments of carbamoyl phosphate synthase-1 (CPS-1), an abundant enzyme of the hepatic urea cycle, which is normally located in the mitochondrial matrix. In addition, we developed a new sandwich immunoassay to determine circulating CPS-1 in human and baboons. We found CPS-1 to be induced by LPS and to be released into the circulation of healthy humans and baboons as early as 4 to 5 h after stimulation. Similarly, CPS 1 levels increased after injection of gram-positive bacteria in another baboon model. Enhanced CPS-1 levels were also detected in serum of patients with sepsis. Our data demonstrate fragmentation of CPS-1 in the liver and early increase in circulating CPS-1 levels under septic conditions. We suggest that circulating CPS 1 might serve as a novel serum marker indicating mitochondrial impairment of the liver and/or the small intestine in critically ill patients. PMID- 15897807 TI - Administration of recombinant interleukin-11 improves the hemodynamic functions and decreases third space fluid loss in a porcine model of hemorrhagic shock and resuscitation. AB - We have previously demonstrated that the administration of recombinant human interleukin-11 (rhIL-11) during resuscitation improves the blood pressure in a rodent model of hemorrhagic shock. The purpose of this study was to determine whether the effects of rhIL-11 could be reproduced in a large animal model and to elucidate the impact of rhIL-11 administration on the intravascular volume status and the degree of third space fluid loss after resuscitation. A 40% blood volume hemorrhage was induced in swine (n = 45, weight of 25-35 kg) followed by a 1-h shock period and resuscitation with 0.9% sodium chloride (three times the shed blood volume). The animals were randomized to receive sham hemorrhage (group I, sham); sham hemorrhage and 50 microg/kg rhIL-11 (group II, sham + IL-11); no drug (group III, saline); or 50 microg/kg rhIL-11 (group IV, IL-11). Blood and urine samples were obtained and analyzed at baseline, at the end of hemorrhaging, and thereafter once every hour. The pleural and peritoneal effusions were precisely quantified by using clinically accepted criteria. The mean arterial pressure (MAP) was higher postresuscitation (PR) in groups I, II, and IV (71.4 +/- 7.5 mmHg, 71.0 +/- 8.9 mmHg, and 72.9 +/- 12.3 mmHg, respectively) than in group III (59.9 +/- 10.9 mmHg), and the cardiac output of PR was higher in group IV (3.46 +/- 0.56 L/min) than in group III (2.99 +/- 0.62 L/min; P < 0.01). The difference in MAP between groups I and II became statistically significant at 40 min after rhIL-11 injection and such a difference persisted for 90 min. After resuscitation, the urine output was higher, and the urine specific gravity and third space fluid loss were lower in group IV (1434 +/- 325 mL and 1.0035, 82 +/- 21 mL) than in group III (958 +/- 390 mL and 1.0053, 125 +/- 32 mL; P < 0.05). In a porcine model of hemorrhagic shock, the administration of rhIL-11 at the start of resuscitation significantly improved the cardiac output and blood pressure. This strategy also significantly reduced the extent of third space fluid losses while also having a favorable impact on the intravascular volume status as evidenced by the improved urine output. PMID- 15897808 TI - Growth differentiation factor-15/macrophage inhibitory cytokine-1 induction after kidney and lung injury. AB - The immunoregulatory cytokine macrophage inhibitory cytokine-1 (MIC-1), a divergent TGF-beta family member, and its murine ortholog, growth/differentiation factor-15 (GDF-15) are induced in hepatocytes by surgical and chemical injury and heat shock. To better understand the in vivo role this factor plays in organ injury, we examined the regulation of GDF-15 in murine models of kidney and lung injury. We demonstrate herein induction of GDF-15/MIC-1 after surgical, toxic/genotoxic, ischemic, and hyperoxic kidney or lung injury. Gdf15 induction was independent of protein synthesis, a hallmark of immediate-early gene regulation. Although TNF induced GDF-15 expression, injury-elicited Gdf15 expression was not reduced in mice deficient for both TNF receptor subtype. Furthermore, although the stress sensor p53 is known to induce GDF-15/MIC-1 expression, injury-elicited Gdf15 expression was unchanged in p53-null mice. Our results demonstrate that GDF-15 induction after organ injury is a hallmark of many tissues. These data demonstrate that GDF-15/MIC-1 is an early mediator of the injury response in kidney and lung that might regulate inflammation, cell survival, proliferation, and apoptosis in a variety of injured tissues and disease processes. PMID- 15897809 TI - Increase plasma viscosity sustains microcirculation after resuscitation from hemorrhagic shock and continuous bleeding. AB - Resuscitation from hemorrhagic shock (50% of blood volume, BV) followed by continuous bleeding (20% of BV per hour, over the entire observation time, 90 min) was studied in the unanesthetized hamster chamber window model. Blood losses equaled 100% of total BV. A single volume infusion (resuscitation) was performed 60 min after hemorrhage using 25% of the BV with 10% hydroxyethyl starch (HES 200, group HES4), or a mixture of HES 200 with 0.3% or 0.6% (w/v) alginate (groups HES7 and HES10, respectively) leading to solutions with a uniform colloidal oncotic pressure (84-87 mmHg) and viscosities ranging from 3.8 to 9.8 cp. Results showed all solutions to be similar immediately after resuscitation (10-15 min) and diverged after this initial period. The viscosity-enhanced solutions showed improved and longer-lasting effects (90 min) relative to the conventional low viscosity, in terms of sustained arterial blood pressure, microvascular flow, capillary perfusion, and laboratory parameters. All microvascular parameters 90 min after resuscitation with low viscosity fell back to the shock level. Improved recovery obtained with a hyperviscous plasma expander was related to microcirculation shear stress preservation, leading to improve blood flow by lowering peripheral vascular resistance when compared with low viscosity resuscitation. These findings suggest the possibility of using hyperviscous plasma expanders to prolong the period for initial treatment of blood losses and definitive institution therapy. PMID- 15897810 TI - Thalidomide treatment reduces colon injury induced by experimental colitis. AB - The immunological and genetic pathogeneses of inflammatory bowel disease (IBD) have been well studied but not well elucidated in the recent years. Accordingly, the pharmacological treatment of IBDs is focusing upon the individual pathologic step (targeting therapy). It has been shown recently that new drugs such as biological immunomodulating agents and anti-inflammatory cytokines have better short-term effects in some respects than the conventional drugs, and they might change the treatment strategy of IBDs in the near future. The aim of the present study was to examine the effects of thalidomide treatment in the development of experimental colitis. To address this question, we used an experimental model of colitis, induced by dinitrobenzene sulfonic acid (DNBS). DNBS-treated mice experienced diarrhea and weight loss. At 4 days after administration of DNBS, the mucosa of the colon exhibited large areas of necrosis. The observed mucosa alteration was associated with the colon production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and vascular endothelial growth factor (VEGF). Neutrophil infiltration (determined by histology as well as an increase in myeloperoxidase activity in the mucosa) was associated with an upregulation of intercellular adhesion molecule-1. Immunohistochemistry for nitrotyrosine and poly (ADP ribose) showed an intense staining in the inflamed colon. When compared with DNBS-treated mice, thalidomide-treated (200 mg/kg orally) mice subjected to DNBS-induced colitis experienced a significantly lower rate in the extent and severity of the histological signs of colon injury. Thalidomide also caused a substantial reduction of the rise in myeloperoxidase activity (mucosa), in the increase in the tissue levels of TNF-alpha, IL-1beta, and VEGF, in the increase in staining (immunohistochemistry) for nitrotyrosine and for poly (ADP ribose), as well as in the upregulation of intercellular adhesion molecule-1 caused by DNBS in the colon. Thus, thalidomide treatment reduces the degree of colitis caused by DNBS. We propose that this evidence may help to clarify the therapeutic actions of thalidomide in patients with Crohn's disease. PMID- 15897811 TI - Evidence of transcellular albumin transport after hemorrhagic shock. AB - Hemorrhagic shock-induced ischemia-reperfusion injury is characterized by an increase in microvascular permeability. This increase in permeability is thought to occur mainly via passive transport through interendothelial cell junctions. However, recent data have suggested that a transcellular (caveolae) transport mechanism(s) may also play a role after shock. The purpose of our study was to investigate the role of caveolae transport after hemorrhagic shock. After a control period, blood was withdrawn to reduce the mean arterial pressure to 40 mmHg for 1 h in urethane-anesthetized Sprague-Dawley rats. Mesenteric postcapillary venules in a transilluminated segment of small intestine were examined to determine changes in permeability. Rats received an intravenous injection of fluorescein isothiocyanate-bovine albumin during the control period. The fluorescent light intensity emitted from the fluorescein isothiocyanate bovine albumin was recorded with digital microscopy within the lumen of the microvasculature and was compared with the intensity of light in the extravascular space. The images were downloaded to a computerized image analysis program that quantitates changes in light intensity. This change in light intensity represents albumin extravasation. Our results demonstrated a marked increase in albumin leak after hemorrhagic shock that was significantly attenuated with two different inhibitors of transcellular transport, N ethylmaleimide and methyl-beta-cyclodextrin. These data suggest that caveolae transport plays a significant role in microvascular permeability after hemorrhagic shock. PMID- 15897812 TI - Effects of primary and secondary intra-abdominal hypertension on mesenteric lymph flow: implications for the abdominal compartment syndrome. AB - Intra-abdominal hypertension leading to abdominal compartment syndrome complicates trauma resuscitation. The purpose of this study was to determine the effect of primary (1 degrees) and secondary (2 degrees) intra-abdominal hypertension (IAH) on hemodynamics, intestinal fluid balance, and mesenteric lymph flow. Anesthetized dogs were instrumented with vascular catheters, intra abdominal manometer, and mesenteric lymphatic fistulae. 1 degrees IAH was created by infusing 0.9% saline into the peritoneal cavity to increase abdominal pressure. 2 degrees IAH was created by elevating the inferior vena cava (IVC) pressure between 20 and 25 mmHg and crystalloid resuscitation to create intestinal edema to induce IAH. At baseline and at 30-min intervals, hemodynamics, lymph flow (QL), IVC, and intra-abdominal pressures were measured. Tissue water was determined using microgravimetry to assess gut edema. Results are reported as mean +/- SEM, with n = 7-8 dogs per group. 1 degrees IAH significantly increased CVP and decreased QL. 1 degrees IAH stopped mesenteric QL, thus transvascular fluid flux necessarily exceeded QL, contributing to gut edema formation. 2 degrees IAH significantly increased CVP and QL. 2 degrees IAH increased QL despite elevated IAP. Interstitial protein washdown maintained the plasma-to-interstitial oncotic gradient, thus increased transvascular fluid flux was due principally to increased capillary pressure. Transvascular fluid flux exceeded QL as manifested by increasing gut tissue water as QL plateaued. Modest elevations in IAP significantly affect mesenteric QL and the development of gut edema. The principle of early abdominal decompression to reduce mesenteric/IVC venous hypertension and capillary pressure is supported by these data. PMID- 15897813 TI - The role of calcium desensitization in vascular hyporeactivity and its regulation after hemorrhagic shock in the rat. AB - The objectives of the present study were to investigate the role of calcium desensitization in vascular hyporeactivity, and the regulatory effects of Rho kinase, protein kinase C (PKC), and protein kinase G (PKG) on calcium sensitivity. The vascular reactivity and calcium sensitivity with superior mesenteric artery (SMA) from hemorrhagic shock rat were observed by measuring the contraction initiated by norepinephrine (NE) and Ca2+ under depolarizing conditions (120 mmol/L K) in an isolated organ perfusion system. Angiotensin II (Ang-II) and Fasudil, the Rho-kinase agonist and inhibitor, phorbol 12-myristate 13-acetate (PMA) and staurosporine, the PKC agonist and inhibitor, 8Br-cGMP and KT-5823, the PKG agonist and inhibitor, and Calyculin A, myosin light chain phosphatase (MLCP) inhibitor were used as tool agents. The results indicated that vascular reactivity and calcium sensitivity were decreased after hemorrhagic shock. The cumulative dose-response curve of SMA to NE and Ca2+ after shock was shifted to the right. Ang-II (10 mol/L) could improve the decreased vascular reactivity by increasing the calcium sensitivity of SMA, and insulin (100 nmol/L) could further decrease the vascular reactivity by decreasing the calcium sensitivity of SMA. These results suggested that the vasculature after shock was desensitized to calcium, which played an important role in the onset of vascular hyporeactivity after shock. PMA and KT-5823 could increase the sensitivity of SMA to Ca2+ and made the cumulative dose-response curve shift to the left. In contrast, Fasudil, staurosporine, and 8Br-cGMP decreased the sensitivity of SMA to Ca2+ and made the cumulative dose-response curve of Ca2+ shift to the right. Calyculin A (10 mol/L) pretreatment further enhanced Ang-II, and PMA induced increase of calcium sensitivity, yet weakened the 8Br-cGMP-induced decrease of calcium sensitivity. Taken together, the data suggest that Rho-kinase, PKC, and PKG are involved in the regulation of calcium sensitivity of vascular smooth muscle after hemorrhagic shock, and their regulatory effects on calcium sensitivity of vasculature are possibly related to MLCP. PMID- 15897814 TI - The use of methylene blue in the treatment of anaphylactic shock induced by compound 48/80: experimental studies in rabbits. AB - In this study, the isolated use of methylene blue (MB) in the treatment of anaphylactic shock induced by Compound 48/80 (C48/80), a potent histamine releaser, was examined, and the study of the effects of MB on the function of the aorta artery endothelium was accomplished in vitro. MB was used in a single 3.0 mg/kg dose, and C48/80 was used in a single 4.5 mg/kg dose. The study protocol included the following experimental groups, containing six animals each: group I (control), animals in the absence of any drug action; group II (MB), MB infusion; Group III (C48/80), anaphylactic shock induced by using C48/80; group IV (C48/80 + MB), anaphylactic shock treated with MB infusion at the moment of major hypotension; and group V (MB + C48/80), prevention of anaphylactic shock with MB by means of MB infusion minutes before the 4.5 mg/kg C48/80 infusion. Nitric oxide plasma levels were measured in each of the experimental groups. After the in vivo studies were performed, an in vitro study was conducted using segments of the abdominal aortas of the rabbits to determine the effect of MB on the arterial endothelium. The results obtained in the present investigation have shown that MB intravenous infusion does not change the mean arterial pressure when compared with the control group (n = 6 in each group, P < 0.05); that C48/80 is effective in producing experimental anaphylactic shock (n = 6, P < 0.05); that the attempt to prevent anaphylactic shock with MB results in a mean prolongation of animal survival ranging from 17 to 34 min (n = 6 in each group, P < 0.05); that MB is effective in reversing anaphylactic shock in all the studied rabbits (n = 6, P < 0.05); that absolute and percentage plasma nitrate values obtained with the experimental groups do not differ (n = 6, each group, P < 0.05); and that the in vitro study of segments of abdominal aorta has shown that there has not been endothelial dysfunction in any of the groups (n = 6 in each group, P < 0.05). The good results obtained in this study open a research path that may offer data to define new paradigms for treating anaphylaxis. PMID- 15897815 TI - An in vitro animal study of the biomechanical responses of anulus fibrosus with aging. AB - STUDY DESIGN: In vitro biomechanical tests were performed on aged (group A) and young (group B) porcine intervertebral discs. OBJECTIVES: The in vitro biomechanical responses of aged and young porcine intervertebral discs were measured under designated axial compressive loads and analyzed. SUMMARY OF BACKGROUND DATA: From the biomechanical point of view, the major biomechanical functions of intervertebral discs are to absorb and distribute external loadings. Although the histological observation of intervertebral discs on the effect of aging and related degeneration has been extensively studied and described, the changes in those biomechanical functions attributable to aging are still left to be studied. METHODS: Two groups were set for mechanical tests. Group A consisted of 24 motion segments obtained from female porcine lumbar (44.0 +/- 2.8-months old). The group B consisted of 30 motion segments from female porcine lumbar (6.2 +/- 1.3 months old). The specimens were chosen randomly from all levels. For the measurements of biomechanical responses, a pressure transducer was placed on anterior and posterolateral locations of anulus fibrosus. Morphological and histological observations were carried out to confirm any age-related changes in both groups. Intradiscal pressures and relaxation times were measured and calculated at points in the anulus fibrosus under designated axial compressive loads. RESULTS: Morphological and histological difference between group A and group B were confirmed with H&E staining and other measurements. Group A showed a lower ratio of nucleus pulposus area to total disc area than did group B. There was no significant difference in the intradiscal pressure between groups as measured in the anterior zone, except at an axial load of 740 N. However, a significant pressure difference was found in the posterolateral zone when the axial load was 542 N or greater (P < 0.05). At 740 N, the average relaxation time for group A was significantly longer than that for group B (P < 0.05). CONCLUSIONS: Differences in biomechanical responses between groups were confirmed. Group A was less flexible and slower at energy relaxation under axial loading. A larger proportion of the external load was taken by the posterolateral part of the degenerative discs. These results were consistent with clinical experiences: 1) most hernias are observed more often at the posterolateral side than other sides, and 2) the degeneration attributable to age reduces the function of absorbing and distribution of external loadings. PMID- 15897816 TI - Spinal cord infarction following cervical transforaminal epidural injection: a case report. AB - STUDY DESIGN: Case report. OBJECTIVE: A review of the literature about spinal cord infarction with epidural steroid injections and report of one case. SUMMARY OF BACKGROUND DATA: A 53-year-old man with a history of chronic cervical pain and multilevel degenerative disc disease with multiple posterior disc protrusions on cervical imaging. The patient received a left C6 tranforaminal injection for therapeutic pain relief, with fluoroscopic confirmation of left C6 nerve root sheath spread of injectable contrast. Approximately 10 to 15 minutes post procedure, he noted weakness in his left arm and bilateral lower limbs. Initial cervical magnetic resonance imaging revealed no cord signal change, but a follow up study 24 hours later demonstrated patchy increased T2 and short tau inversion recovery signal in the cervical cord from the odontoid to C4-C5 vertebral levels. This was consistent with a diffuse vascular infarct to the cervical cord, resulting in motor-incomplete tetraplegia. RESULTS: This is one of a few reported cases of spinal cord infarction after cervical epidural injections. No direct cord trauma occurred. Previously reported risk factors of spinal infarction, such as hypotension and large injectate volumes, were noncontributory in this case. CONCLUSIONS: Cervical epidural injections, despite careful localization, carry a risk of vascular infarction to the spinal cord, even in the absence of direct cord trauma. The etiology of these infarctions and identifying those patients at risk remain uncertain. PMID- 15897817 TI - A child who recovered completely after spinal cord injury complicated by C2-3 fracture dislocation: case report. AB - STUDY DESIGN: This was a case of a child who recovered completely after spinal cord surgery complicated by C2-C3 fracture dislocation. OBJECTIVES: To clarify the important issue with regard to the diagnosis and treatment of possible spinal cord injury complicated by C2-C3 fracture dislocation in children. SUMMARY OF BACKGROUND DATA: Reports on spinal cord injury complicated by C2-C3 fracture dislocation in children who recovered completely after surgery are very rare. Moreover, there are no reports of cases in which described MRI and magnetic resonance angiography obtained 5 years after surgery. METHODS: A 4-year-old girl with complete fracture dislocation of C2-C3 and spinal cord injury of Frankel B because of a traffic injury. A middle incision was made in a prone position under general anesthesia, and C2-C3 was fixed with interlaminal wiring according to the McGraw modified method. RESULTS: The postoperative course was excellent, and the patient recovered completely. An MRI performed 5 years after surgery revealed no definitive abnormality in spinal cord and patency of vertebral arteries. CONCLUSION: This was a very rare case of spinal cord injury complicated C2-C3 fracture dislocation in children. The present case is of interest in that it demonstrated the possibility of recovery in a child from spinal cord injuries of Frankel B immediately after injury, if complex injuries in multiple organs are controlled by systemic management. PMID- 15897818 TI - Triple total cervical vertebrectomy for a giant cell tumor: case report. AB - STUDY DESIGN: Case report of a triple total cervical vertebrectomy. OBJECTIVE: To describe a new management for cervical tumor. Preoperative planning using arteriography, successive occlusion of both vertebral arteries, and the cervical vertebrectomy are reported. SUMMARY OF BACKGROUND DATA: Thoracic or lumbar complete vertebrectomy for primary malignant tumor or metastasis is a well established surgical technique. The presence of the vertebral arteries appears to have prevented the previous use of complete vertebrectomy in the cervical spine. METHODS: A 25-year-old male patient who had a giant cell tumor in C6 underwent hemi-vertebrectomy. Before this surgical procedure, the ipsilateral vertebral artery was embolized. The tumor recurred locally 18 months later. Using temporary balloon occlusion of the remaining vertebral artery, an abundant collateral circulation from the cervical arteries to the vertebrobasilar territory was shown. Triple total cervical vertebrectomy from C5-C7 was then performed with double stage surgery. RESULTS: At 2-year follow-up, the patient is tumor-free. CONCLUSIONS: Complete resection of malignant cervical vertebrae is possible if both vertebral arteries can be successively occluded, permitting complete removal of the transverse processes. PMID- 15897819 TI - Point of view. PMID- 15897820 TI - Cerebrospinal fluid fistula secondary to dural tear in anterior cervical discectomy and fusion: case report. AB - STUDY DESIGN: A case of cerebrospinal fistula secondary to a dural tear during anterior cervical discectomy and fusion. OBJECTIVES: To report a quite rare complication associated with anterior cervical discectomy and remind the spinal surgeons that this infrequent complication can easily become a very serious one. SUMMARY OF BACKGROUND DATA: Anterior cervical discectomy represents one of the most commonly performed spinal procedures. Of the associated complications, accidental dural tear can lead to the development of a cerebrospinal fluid (CSF) fistula. Although this complication has been mentioned in several clinical series, the body of knowledge regarding incidence and appropriate treatment is definitely limited. METHODS: After undergoing anterior cervical discectomy and fusion for an extruded disc at the C4-C5 level, a CSF fistula developed in a 37 year-old patient as a result of a dural tear. The patient underwent a second procedure for surgical wound revision, meticulous dural opening coverage, and insertion of a lumbar drain for draining CSF for 5 days. RESULTS: The patient was hospitalized for 5 days and then discharged with no evidence of CSF leakage. His follow-up of 9 months revealed complete resolution of his preoperative symptomatology and no other problems associated with the complication of the CSF fistula. CONCLUSION: Early identification of this complication and aggressive treatment with insertion of lumbar drain, CSF drainage for 4-5 days, and coverage of the dural tear with fibrin sealant or autologous fascia graft can prevent the development of any consequences. PMID- 15897822 TI - Cost-effectiveness of combined manipulation, stabilizing exercises, and physician consultation compared to physician consultation alone for chronic low back pain: a prospective randomized trial with 2-year follow-up. AB - STUDY DESIGN: A prospective, randomized controlled trial. OBJECTIVE: To examine long-term effects and costs of combined manipulative treatment, stabilizing exercises, and physician consultation compared with physician consultation alone for chronic low back pain (cLBP). SUMMARY OF BACKGROUND DATA: An obvious gap exists in knowledge concerning long-term efficacy and cost-effectiveness of manipulative treatment methods. METHODS: Of 204 patients with cLBP whose Oswestry Disability Index (ODI) was at least 16%, 102 were randomized into a combined manipulative treatment, exercise, and physician consultation group (i.e., a combination group), and 102 to a consultation alone group. All patients were clinically examined, informed about their back pain, and encouraged to stay active and exercise according to specific instructions based on clinical evaluation. Treatment included 4 sessions of manual therapy and stabilizing exercises aimed at correcting the lumbopelvic rhythm. Questionnaires inquired about pain (visual analog scale (VAS)), disability (ODI), health-related quality of life (15D Quality of Life Instrument), satisfaction with care, and costs. RESULTS: Significant improvement occurred in both groups on every self-rated outcome measurement. Within 2 years, the combination group showed only a slightly more significant reduction in VAS (P = 0.01, analysis of variance) but clearly higher patient satisfaction (P = 0.001, Pearson chi2) as compared to the consultation group. Incremental analysis showed that for combined group compared to consultation group, a one-point change in VAS scale cost $512. CONCLUSIONS: Physician consultation alone was more cost-effective for both health care use and work absenteeism, and led to equal improvement in disability and health-related quality of life. It seems obvious that encouraging information and advice are major elements for the treatment of patients with cLBP. PMID- 15897823 TI - The influence of alendronate treatment and bone graft volume on posterior lateral spine fusion in a porcine model. AB - STUDY DESIGN: An experimental animal study with randomized, paired control design was conducted using a porcine model. OBJECTIVES: The aim of this study was to evaluate the influence of alendronate treatment and the significance of different amounts of bone graft on posterior lateral spine fusion. SUMMARY OF BACKGROUND DATA: Treatment with bisphosphonates inhibits osteoclast-induced bone resorption and increases bone quality and density. It has been widely used clinically for treatment of osteoporosis. Bisphosphonates have been reported to elongate the callus remodeling process during fracture healing. Bisphosphonate treatment may modify bone graft healing and the remodeling process in spine fusion. The bone resorption phase exists during the healing process. Extensive bone graft resorption could reduce the basis for new bone formation, which could be an important factor for failure of spine fusion. Furthermore, different amounts of initially applied bone graft may influence spine fusion rate and bone graft incorporation process. METHODS: Twenty-two pigs were included in the study. Eleven pigs in the treatment group received alendronate 10 mg/day p.o. for 3 months after surgery. The other 11 pigs received no bisphosphonate and served as control group. Posterior lateral fusion with the CD Horizon system was performed on the lumbar spine using different amounts of autograft (4 g on one side and 8 g on the other side) in all animals. The fusion was evaluated using radiograph, CT scan, and histomorphometry at 3 months after operation. RESULTS: There was no statistical difference in either fusion rate or fusion mass volume between the two groups. The fusion rate based on radiograph was 75% on the 8-g autograft side and 45% on the 4-g side (P < 0.05). The mean volume of fusion mass was 2.36 cm3 on the 4-g side and 3.29 cm3 on the 8-g side (P < 0.01). No difference was found in either trabecular bone volume or fusion rate between treatment and control groups using histologic evaluation.The treatment group showed a higher fibrous tissue volume(P < 0.05), higher proportion of woven bone structure(P < 0.001), and lower bone marrow volume (P = 0.088) in the fusion mass. Different amounts of bone graft did not change the tissue composition of the fusion mass. CONCLUSIONS: Alendronate treatment in this study decreased fusion mass remodeling without inhibiting fusion rate. Increased amounts of autologous bone graft could improve the fusion rate in this experimental spine fusion study. PMID- 15897824 TI - Matrix remodeling expression in anulus cells subjected to increased compressive load. AB - STUDY DESIGN: Mechanobiology study of gene expression changes as a result of compressive overload of anular fibrochondrocytes. OBJECTIVE: To test hypotheses regarding phenotype shift in genes coding for representative extracellular matrix (ECM) proteins and matrix modulators. SUMMARY OF THE BACKGROUND DATA: In degenerative disc disease, the transfer of compressive load through the disc shifts largely from the nucleus onto the anulus. In vivo models simulating this condition have shown derangement of the collagenous ultrastructure in the anulus. In vitro models of cultured anulus cells subjected to static compressive stress generally suggest a down-regulation of synthesis. This study evaluated the expression of specific isomers of genes responsible for mechanical viability and metabolism of the disc under cyclic compressive loads. METHODS: Fibrochondrocytes were digested from the anuli of 3, 2-week-old pigs, embedded in 1.5% alginate gel, and hydrostatically compressed at 0.5 Hz for 3 hours to amplitudes of 10 and 30 atm. These levels represented nominal load transfer through the healthy disc and high load transfer through the degenerative disc. Ribonucleic acid was isolated, reverse transcribed, and evaluated by real-time polymerase chain reaction for expression of type I (C-I) and type II (C-II) collagen, aggrecan, the matrix metalloproteinase (MMP-1), and the transforming growth factor beta (TGFbeta-1). Results were expressed at percentages of uncompressed controls. RESULTS: The lower pressure of 10 atm resulted in up-regulation of all ECM protein genes. C-I and C-II both averaged 141%, and aggrecan 121% of controls (P < 0.05). MMP-1 and TGFbeta-1 were essentially unchanged. With the pressure increased to 30 atm, C-II remained approximately at the level expressed under lower pressure, but C-I was reduced to 42% of controls (P < 0.05), indicating a phenotype shift. MMP-1 and TGFbeta-1 also were down-regulated to 71% and 54% of controls, respectively (P < 0.05). CONCLUSIONS: The up-regulation of the ECM genes with nominal pressure highlights the mechanobiological importance of common activity in fibrocartilage homeostasis. Differential regulation of the 2 primary collagen types with high pressure indicates a capacity of the anulus to remodel according to pathomechanical conditions. PMID- 15897825 TI - Lower dose of rhBMP-2 achieves spine fusion when combined with an osteoconductive bulking agent in non-human primates. AB - STUDY DESIGN: A non-human primate lumbar intertransverse process arthrodesis model was used to evaluate the effectiveness of different formulations of recombinant human bone morphogenetic protein-2 (rhBMP-2) to induce consistent bone formation. OBJECTIVE: To determine if the combination of rhBMP-2/absorbable collagen sponge (ACS) wrapped around a bulking agent, consisting of a biphasic calcium phosphate/collagen composite, could achieve posterolateral spine fusion with a dose of rhBMP-2 (3.0 mg/side) that previously failed to induce posterolateral fusion in rhesus monkeys with other carriers. SUMMARY OF BACKGROUND DATA: Successful bone induction in both human and non-human primates has required higher concentrations of BMP than were required in lower order models. The Food and Drug Administration approved concentration of rhBMP-2 for interbody fusion (1.5 mg/mL) when delivered on the ACS alone without a bulking agent (doses 3-9 mg/side) has failed to induce clinically relevant amounts of bone formation in a posterolateral spine fusion model in rhesus monkeys. Previously, a higher concentration of 2.0 mg/mL of rhBMP-2 delivered on stacked sheets of a biphasic calcium phosphate ceramic/collagen compression resistant matrix (CRM) was required to achieve fusion in the rhesus monkey and was the basis for this study (doses of 6-12 mg/side). METHODS: Nine skeletally mature, rhesus macaque monkeys underwent single level posterolateral arthrodesis at L4 L5. Two different rhBMP-2 doses were evaluated in 3 delivery configurations. The first 3 monkeys received 10 mg/side (2.5 mL at 4.0 mg/mL) of rhBMP-2 loadeddirectly onto a CRM carrier (15% hydroxyapatite/85%beta-tricalcium phosphate ceramic/collagen matrix), resulting in a final concentration of 2.0 mg/mL. The second 3 monkeys received 3 mg/side (2.0 mL at 1.5 mg/mL) of rhBMP-2 loaded directly on the CRM carrier, resulting in a 0.6 mg/mL final concentration. Three additional monkeys also received the 3 mg/side (2.0 mL at 1.5 mg/mL) of rhBMP-2 delivered on an ACS, which was then wrapped around the dry CRM matrix used as a bulking agent, yielding a 1.5 mg/mL final concentration of rhBMP-2 on the sponge wrapped around the bulking agent. The monkeys were euthanized at 24 weeks after surgery. Manual palpation, plain radiographs, computerized tomography, and nondecalcified histology were used to evaluate fusion in a blinded fashion. RESULTS: The 3 monkeys with 10 mg rhBMP-2 placed directly on the CRM carrier (2.0 mg/mL final concentration) achieved solid fusion. The 3 monkeys that underwent fusion with 3 mg of rhBMP-2 placed directly on the CRM carrier (0.6 mg/mL final concentration) failed to achieve fusion. In contrast, the 3 monkeys that underwent fusion with the same 3 mg dose of rhBMP-2 dispensed only on an ACS that was wrapped around the CRM achieved solid bilateral fusion. CONCLUSIONS: This study shows the importance of carrier optimization and final implant protein concentration for the successful delivery of rhBMP-2. By combining the properties of the ACS with the CRM, the required dosage of rhBMP-2 was diminished by more than 3-fold in the non-human primate model. This finding suggests that the currently available concentration of rhBMP-2 (1.5 mg/mL) could be successful for achieving posterolateral spine fusion when combined with an osteoconductive bulking agent that can support the induced new bone formation. PMID- 15897826 TI - The use of cultured bone marrow cells in type I collagen gel and porous hydroxyapatite for posterolateral lumbar spine fusion. AB - STUDY DESIGN: Posterolateral lumbar transverse process fusion was completed using the cultured bone marrow cells in type I collagen gel and porous hydroxyapatite. OBJECTIVE: To compare the efficacy of cultured bone marrow cells with that of bone morphogenetic protein (BMP) as a graft alternative to autologous bone for posterolateral spine fusion. SUMMARY OF BACKGROUND DATA: The clinical application of BMP for spinal fusion may be limited by high dose and cost. Recently, mesenchymal stem cells have been studied in various fields because of their capability to differentiate into various cells, including those in the osteogenic lineage. METHODS: Thirty adult rabbits were used. Each underwent single-level, bilateral, posterolateral intertransverse process fusions at L4-L5. The animals were divided into 4 groups, each according to the material implanted: (1) autologous bone (autograft, n = 9); (2) porous hydroxyapatite (HA) particles and type I collagen sheet with 100 microg rhBMP-2 (BMP-HA, n = 7); (3) bone marrow cells (1 x 10(6) cells/mL, low-marrow-HA, n = 7); and (4) bone marrow cells (1 x 10(8) cells/mL, high-marrow-HA, n = 7). Before implantation for groups 3 and 4, fresh bone marrow cells from the iliac crest of each animal were cultured in a standard medium for 2 weeks. For one additional week, the marrow cells were cultured in 10(-8) M dexamethasone, type I collagen gel, and HA. Animals were euthanized 6 weeks after surgery. Spinal fusions were evaluated by radiograph, manual palpation, and histology. RESULTS: The fusion rates were 4 of 7 in the autograft group, 7 of 7 in the BMP-HA group, 0 of 7 in the low-marrow-HA group, and 5 of 7 in the high-marrow-HA group. The histology in the BMP-HA and high marrow-HA groups showed that grafted HA fragments were connected with mature new bone. The pores of HA fragments were filled up with bone matrix. In the low marrow-HA group, fibrous tissue was predominant in the grafted fragments. CONCLUSIONS: This study shows that the cultured bone marrow cells can act as a substitute for autograft or BMP in spine fusion. The current formulation may yield improved fusion success and better quality of fusion bone as compared to autograft. PMID- 15897827 TI - Human intervertebral disc aggrecan inhibits endothelial cell adhesion and cell migration in vitro. AB - STUDY DESIGN: The effect of human intervertebral disc aggrecan on endothelial cell growth was examined using cell culture assays. OBJECTIVE: To determine the response of endothelial cells to human intervertebral disc aggrecan, and whether the amount and type of aggrecan present in the intervertebral disc may be implicated in disc vascularization. SUMMARY OF BACKGROUND DATA: Intervertebral disc degeneration has been associated with a loss of proteoglycan, and the ingrowth of blood vessels and nerves. Neovascularization is a common feature also of disc herniation. Intervertebral disc aggrecan is inhibitory to sensory nerve growth, but the effects of disc aggrecan on endothelial cell growth are not known. METHODS: Aggrecan monomers were isolated separately from the anulus fibrosus and nucleus pulposus of human lumbar intervertebral discs, and characterized to determine the amount and type of sulfated glycosaminoglycan side chains present. The effects of these aggrecan isolates on the cellular adhesion and migration of the human endothelial cell lines, HMEC-1 and EAhy-926, were examined in vitro. RESULTS: Homogenous substrata of disc aggrecan inhibited endothelial cell adhesion and cell spreading in a concentration dependent manner. In substrata choice assays, endothelial cells seeded onto collagen type I migrated over the collagen until they encountered substrata of disc aggrecan, where they either stopped migrating, retreated onto the collagen, or, more commonly, changed direction to align along the collagen-aggrecan border. The inhibitory effect of aggrecan on endothelial cell migration was concentration dependent, and reduced by enzymatic treatment of the aggrecan monomers with a combination of chondroitinase ABC and keratinase/keratinase II. Anulus fibrosus aggrecan was more inhibitory to endothelial cell adhesion than nucleus pulposus aggrecan. However, this difference did not relate to the extent to which the different aggrecan isolates were charged, as determined by colorimetric assay with 1,9-dimethylmethylene blue, or to marked differences in the distribution of chondroitin sulfated and keratan sulfated side chains. CONCLUSIONS: Human intervertebral disc aggrecan is inhibitory to endothelial cell migration, and this inhibitory effect appears to depend, in part, on the presence of glycosaminoglycan side chains on the aggrecan monomer. PMID- 15897828 TI - Endoscopic mechanical spinal hemiepiphysiodesis modifies spine growth. AB - STUDY DESIGN: An in vivo porcine model of progressive scoliosis as an inverse analog of a proposed method of early surgical treatment. OBJECTIVES: To test the hypothesis that scoliotic curvatures may be repeatedly created using anatomically based vertebral staples and thoracoscopic surgical procedures. SUMMARY OF BACKGROUND DATA: Staple hemiepiphysiodesis is an established method for treating knee deformities. Similar procedures have so far failed to arrest or correct deformities of the spine. While experimental studies continue to suggest that spine growth is modifiable, no prior clinically translatable method has been shown to clearly and consistently alter vertebral growth. METHODS: Custom spine staples were implanted into midthoracic vertebrae of seven skeletally immature normal pigs. Each staple spanned an intervertebral disc and two growth plates and was fixed to adjacent vertebrae with screws. The animals were anesthetized biweekly for radiography during the 8-week study period. Final radiographs were taken after spine harvest. Initial and final postoperative Cobb angles were compared statistically. RESULTS: Five animals completed the protocol with a weight increase of 142% in 8 weeks. Coronal plane curvatures increased significantly with time, from 0.8 (+/-1.8) to 22.4 (+/-2.8; P = 0.0001). On average, sagittal plane curvatures did not increase with time. CONCLUSIONS: Spinal hemiepiphysiodesis using an anatomically based implant and minimally invasive procedures repeatedly induced spine curvature in a normal porcine model. These techniques may slow, and perhaps even correct, early progressive spine deformity without long rod instrumentation or fusion. PMID- 15897829 TI - Neuroprotection and enhanced recovery with edaravone after acute spinal cord injury in rats. AB - STUDY DESIGN: The effect of edaravone, a novel free radical scavenger, was assessed functionally and histologically using a rat spinal cord contusion model. OBJECTIVE: To investigate the effect of edaravone on neuroprotection after spinal cord injury in rats. SUMMARY OF BACKGROUND DATA: The spinal cord injury results in immediate physical damage (primary injury), followed by a prolonged neural tissue disorder (secondary injury). This secondary injury process has been suggested to be induced by lipid peroxidation. Edaravone has been reported to inhibit lipid peroxidation in cerebral ischemia models. METHODS: Spinal cord injury at the T10 level was induced with a weight drop device (10 g weight, 25 mm height). Edaravone was administered intravenously as a bolus dose of 5 mg/kg at 5 minutes, 24 hours, and 48 hours after injury (edaravone-treated rats). In control rats, nothing was administered. Functional assessment was conducted weekly using the Basso-Beattie-Bresnahan locomotor rating scores. Histologically, a percentage of spared white matter area was calculated. The effects of intravenous administration of edaravone on lipid peroxide formation in rat spinal cord homogenate were examined using the thiobarbituric acid test for malonyldialdehyde production. RESULTS: Six weeks after injury, edaravone-treated rats showed significantly higher motor score and larger spared white matter area than control rats. The administration of edaravone attenuated malonyldialdehyde production in spinal cord homogenate by >45%. CONCLUSION: Edaravone enhanced functional recovery and preserved more spinal cord tissue after spinal cord injury in rats. The attenuation of posttraumatic lipid peroxide formation by edaravone partially contributed to this enhancement. PMID- 15897830 TI - Abnormal motion in spondylolytic spondylolisthesis. AB - STUDY DESIGN: A retrospective, cohort study of the kinematics of the lumbar spine of patients with spondylolisthesis compared with asymptomatic normal subjects. OBJECTIVE: To determine if abnormal motion could be detected in the radiographs of patients with spondylolisthesis. SUMMARY OF BACKGROUND DATA: Contrary to the prevailing conviction that lumbar segments affected by lytic spondylolisthesis are unstable, multiple studies have failed to find evidence of increased or abnormal motion at these segments. However, these studies did not use techniques that might reveal abnormalities in the quality of motion, as opposed to its magnitude. METHODS: The flexion-extension radiographs of 13 patients with spondylolytic spondylolisthesis were analyzed to determine the location of their instantaneous centers of rotation, and their magnitudes of translation and sagittal rotation. Normative data were obtained by applying the same techniques to the radiographs of 20 asymptomatic subjects. RESULTS: All but 1 of the 13 patients had at least one segment with abnormal motion. Only one patient had excessive translation at the lytic segment. Five patients had minor abnormalities affecting either the lytic segment or ones above, and 6 had paradoxical motion at the lytic segment in which the center of rotation was located above L5, instead of below, and in which L5 translated backward instead of forward during flexion. CONCLUSIONS: A proportion of patients with spondylolisthesis had highly abnormal movements but ones with normal magnitudes of motion. Determining the instantaneous axes of rotation reveals the abnormal quality of motion. PMID- 15897831 TI - Adjacent level intradiscal pressure and segmental kinematics following a cervical total disc arthroplasty: an in vitro human cadaveric model. AB - STUDY DESIGN: In vitro investigation of cervical adjacent level intradiscal pressures (IDPs) following a total disc replacement arthroplasty. OBJECTIVES: The current in vitro study was undertaken to compare adjacent level IDPs and operative level kinematics following a cervical arthroplasty versus an arthrodesis procedure. SUMMARY OF BACKGROUND DATA: Clinical data indicate the incidence of symptomatic transition syndrome to be as high as 3% annually following a cervical interbody arthrodesis. Recent developments in the motion preservation technology should, in theory, minimize transition syndrome at the adjacent levels. METHODS: A total of 10 human cadaveric cervical spines were used in this investigation. Following intact analysis, all specimens were sequentially reconstructed at C5-C6 with 1) total disc replacement (TDR), 2) allograft dowel, and 3) allograft dowel + anterior cervical plate. Testing was performed in displacement control under axial rotation, flexion/extension, and lateral bending loading modes. IDPs were recorded at C4-C5 and C6-C7 whereas peak range of motion (ROM) and NZ were monitored at C5-C6 level. RESULTS: Similar IDPs were recorded between the intact condition and a TDR reconstruction at both adjacent levels under all loading modes (P > 0.05). However, the C4-C5 IDP values produced under flexion/extension testing for both arthrodesis treatments were significantly higher than the means obtained for the intact and disc replacement groups (P < 0.05). Similar intergroup differences were observed at the C6-C7 level; however, statistical significance was achieved during all three loading methods (P < 0.05). C5-C6 ROM analysis indicated a significantly lower ROM for both arthrodesis constructs compared with intact and TDR groups during flexion/extension testing (P < 0.05). No differences were recorded between the intact and the total disc replacement group under any loading conditions (P > 0.05). CONCLUSION: This is a first study to document that a cervical disc replacement arthroplasty procedure maintains adjacent level IDPs and reconstruction level kinematics near the preoperative values. Consequently, total disc replacement may provide an alternative to conventional surgical management of cervical discogenic pathology decreasing the incidence of symptomatic transition syndrome. PMID- 15897832 TI - Magnetic resonance imaging and low back pain in adults: a diagnostic imaging study of 40-year-old men and women. AB - STUDY DESIGN: Cross-sectional cohort study of a general population. OBJECTIVE: To investigate "abnormal" lumbar spine magnetic resonance imaging (MRI) findings, and their prevalence and associations with low back pain (LBP). SUMMARY OF BACKGROUND DATA: The clinical relevance of various "abnormal" findings in the lumbar spine is unclear. Distinguishing between inevitable age-related findings and degenerative findings with deleterious consequences is a challenge. METHODS: Lumbar spine MRI was obtained in 412, 40-year-old individuals. Predefined "abnormal" MRI findings were interpreted without any knowledge of patient symptoms. Associations between MRI abnormalities and LBP were calculated using odds ratios. The "overall picture" of each MRI finding was established on the basis of the frequencies, diagnostic values, and the strength and consistency of associations. RESULTS: Most "abnormal" MRI findings were found at the lowest lumbar levels. Irregular nucleus shape and reduced disc height were common (>50% of individuals). Relatively common (25% to 50%) were hypointense disc signal, anular tears, high intensity zones, disc protrusions, endplate changes, zygapophyseal joint degeneration, asymmetry, and foraminal stenosis. Nerve root compromise, Modic changes, central spinal stenosis, and anterolisthesis/retrolisthesis were rare (<25%). Most strongly associated with LBP were Modic changes and anterolisthesis (odds ratios >4). Significantly positive associations with all LBP variables were seen for hypointense disc signals, reduced disc height, and Modic changes. All disc "abnormalities" except protrusion were moderately associated with LBP during the past year. CONCLUSION: Most degenerative disc "abnormalities" were moderately associated with LBP. The strongest associations were noted for Modic changes and anterolisthesis. Further studies are needed to define clinical relevance. PMID- 15897833 TI - Identification of candidate regions for familial idiopathic scoliosis. AB - STUDY DESIGN: A genomic screen and statistical linkage analysis of 202 families with at least two individuals with idiopathic scoliosis was performed. OBJECTIVES: To identify candidate regions or the autosomal loci that may be involved in the expression of familial idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: A large sample of families with individuals having idiopathic scoliosis (202 families; 1,198 individuals) was ascertained; diagnoses were based on physical examination and radiographic criteria. METHODS: Model-independent linkage analysis of qualitative and quantitative traits (degree of lateral curvature) related to scoliosis was used to screen genotyping data from 391 markers in the 202 families. Subsets of families were determined before genotyping based on the most likely mode of inheritance for each family (autosomal dominant vs. X-linked dominant). Fine mapping results corroborated linkage in the primary candidate regions. RESULTS: Candidate regions on chromosomes 6, 9, 16, and 17 were considered to have the strongest evidence for linkage across all subsets considered. CONCLUSION: Linkage analyses have identified several candidate regions, a significant step in defining the genetic etiology of this disorder. PMID- 15897834 TI - Idiopathic scoliosis in Singapore schoolchildren: a prevalence study 15 years into the screening program. AB - STUDY DESIGN: A point prevalence survey of 72,699 schoolchildren in four age groups was performed. OBJECTIVES: To determine the prevalence rates of idiopathic scoliosis and to compare with a previous prevalence study done 15 years earlier. SUMMARY OF BACKGROUND DATA: Prevalence rates for idiopathic scoliosis of 5 degrees or more in schoolchildren were established in a study performed in 1982. There have been no previous data on prevalence rate changes over time. METHODS: A total of 35,558 boys and 37,141 girls from randomly selected schools were screened for scoliosis. Those with scoliometer readings of more than 5 degrees underwent radiographic evaluation. Prevalence rates were calculated for scoliosis at a predefined Cobb angle of 10 degrees and 5 degrees , the latter for comparison with the previous prevalence study. Curve type and distribution, pubertal status, and symptoms were correlated with the prevalence data. RESULTS: Prevalence rates were 0.05% for girls and 0.02% for boys at 6 to 7 years of age, 0.24% for girls and 0.15% for boys at 9 to 10 years of age, 1.37% for girls and 0.21% for boys at 11 to 12 years of age, and 2.22% and 0.66%, respectively, for girls and boys at 13 to 14 years of age. The ratio of girls to boys increased from 1.6 at 9 to 10 years of age to 6.4 at 11 to 12 years of age. Thoracolumbar curves were the most common (40.1%), followed by thoracic curves (33.3%), double/triple curves (18.7%), and lumbar curves (7.9%). Older children had greater proportions of larger curves. Compared with the previous prevalence study in 1982, there was a significant increase in the prevalence rate in girls 11 to 12 years of age. Screening of 11- to 12- and 13- to 14-year-old girls detected curves in the range suitable for bracing, with nearly 96% and 32% of the age groups, respectively, still amenarche or within a year of menarche, and 57% and 34% of the age groups, respectively, having low Risser grades of 0, 1, and 2. CONCLUSIONS: The overall prevalence rate of idiopathic scoliosis in our school population in 1997 was 0.93% in girls and 0.25% in boys. The prevalence rates were low at 6 to 7 and 9 to 10 years of age but increased rapidly to 1.37% and 2.22% for girls at 11 to 12 and 13 to 14 years of age, respectively. The prevalence rate increased significantly in 11- to 12-year-old girls over a 15 year period from 1982 to 1997. Screening of 11- to 12- and 13- to 14-year-old girls identified a significant number who could benefit from brace treatment. PMID- 15897835 TI - Use of the scoliosis research society outcomes instrument to evaluate patient outcome in untreated idiopathic scoliosis patients in Japan: part I: comparison with nonscoliosis group: preliminary/limited review in a Japanese population. AB - STUDY DESIGN: This preliminary study evaluates untreated Japanese patients with idiopathic scoliosis using the Scoliosis Research Society Outcomes Instrument (SRS-24). OBJECTIVES: To determine the baseline patient outcome score using the SRS-24 for untreated Japanese scoliosis patients compared with a nonscoliosis group. SUMMARY OF BACKGROUND DATA: The SRS instrument with 24 questions was developed to help evaluate patient-perceived outcomes of idiopathic scoliosis treatment. Evaluation of untreated Japanese idiopathic scoliosis patients using the SRS instrument has not been reported. METHODS: Japanese idiopathic scoliosis patients (n = 141) (mean age, 13.6 years; range, 10-17 years) with a Cobb angle of more than 20 degrees who were not treated with a brace or surgery, were evaluated in comparison with a nonscoliosis group (healthy junior high school students; n = 72) using the SRS-24. The scoliosis group was categorized as mild deformity group with a major curve Cobb angle of less than 30 degrees, moderate deformity group with 30 degrees to 49 degrees, and severe deformity group with more than 50 degrees. The patients were evaluated using section 1 (15 questions) of the SRS-24, which was divided into four domains: total pain, general self image, general function, and activity. Reliability, as determined by internal consistency, was validated using Cronbach's alpha for these domain scales. RESULTS: The severe deformity group had the lowest scores compared with the other deformity groups and the nonscoliosis group in pain (P < 0.0001) and self-image (P < 0.05) domains. The scores for questions 3 (P < 0.0001) and 5 (P < 0.0001), evaluation of self-image of back appearance, were significantly lower in the scoliosis group than those in the nonscoliosis group. This tendency was more significant in the patients with greater curve magnitude. Scores for questions 14 and 15, evaluation of general self-image, in the scoliosis group were, however, higher than those in the nonscoliosis group. Internal consistency using Cronbach's alpha was 0.57 (pain), 0.27 (general self-image), -0.08 (general function), and 0.15 (overall level of activity). CONCLUSION: Untreated scoliosis patients with severe deformity were inclined to complain of being self-conscious or distressed regarding their back appearance compared with age- and sex-matched control adolescents. The control group was inclined to have, however, a negative general self-image compared with the patients group. Internal consistency using Cronbach's alpha for all domains was considerably low. The baseline of the SRS-24 scores in the Japanese population might differ from that of the Western population because of differences in national culture. Therefore, further study is necessary to clarify the validity of the SRS-24 domains for Japanese patients. PMID- 15897836 TI - Use of the scoliosis research society outcomes instrument to evaluate patient outcome in untreated idiopathic scoliosis patients in Japan: part II: relation between spinal deformity and patient outcomes. AB - STUDY DESIGN: This study clarifies the relation between the results of the Scoliosis Research Society Outcomes Instrument (SRS-24) and radiographic parameters of back deformity in Japanese idiopathic scoliosis patients. OBJECTIVES: To investigate the relation between magnitude of back deformity and results of the SRS-24 in untreated patients. SUMMARY OF BACKGROUND DATA: In idiopathic scoliosis, it is necessary to clarify the relation between patient perceived outcomes of the deformity and magnitude of back deformity before considering treatment. The relation between the magnitude of spinal deformity and outcomes of untreated patients, however, has not been fully investigated. METHODS: Patients (n = 166) under 30 years of age with untreated scoliosis were evaluated. Radiologic examination included Cobb angle, rotation angle of apical vertebrae, and translation of C7 vertebra from the central sacral line (C7 translation) on the coronal plane. Patient evaluation using section 1 (15 questions) of the SRS-24 was compared with radiologic findings using Spearman's correlation coefficient by rank (rs). RESULTS: The average pain domain score was 27.0 +/- 2.2 points, general self-image 9.9 +/- 1.7 points, general function 12.7 +/- 1.1 points, and overall level of activity 14.9 +/- 0.6 points. In radiologic deformity, the average Cobb angle and rotation angle of the thoracic curve were 35.8 degrees +/- 12.1 degrees (range, 17 degrees-73 degrees) and 13.9 degrees +/- 8.2 degrees (range, 0 degrees-38 degrees), respectively. The average Cobb and rotation angle of the lumbar curve were 31.4 degrees +/- 9.3 degrees (range, 13 degrees-56 degrees) and 15.4 degrees +/- 9.7 degrees (range, 2 degrees-36 degrees), respectively. The mean C7 translation was 12.4 +/- 9.7 mm (range, 0-48 mm). Comparison between individual domains and radiologic measurements revealed that the total pain (rs = -0.33; P < 0.0001) and general self-image (rs = -0.25; P = 0.0024) domain scores had a significant inverse correlation with thoracic curve Cobb angle. Comparison between the scores of individual questions and radiologic measurements revealed that the scores of question 3 (total pain domain) had a significant inverse correlation with thoracic curve Cobb angle (rs = -0.36; P < 0.0001). The scores of question 5 (general self-image domain) had a significant inverse correlation with thoracic curve Cobb angle (rs = -0.41; P < 0.0001) and rotation angle (rs = -0.30; P = 0.0006). CONCLUSION: The patients did not have negative self-image regarding back appearance when the thoracic curve Cobb angle was less than 30 degrees but had a negative self-image when the thoracic curve Cobb angle was more than 40 degrees and the rotation angle was more than 20 degrees. On the other hand, the lumbar curve Cobb angle and the rotation angle did not correlate with patient self-image. The results of the present study will help to define the parameters for the initiation of active treatment and physicians should maintain or reduce scoliotic deformity so that the thoracic curve Cobb angle is less than 40 degrees and the rotation angle is less than 20 degrees in idiopathic scoliosis. PMID- 15897837 TI - Post-traumatic syringomyelia: a review of the cases presenting in a regional spinal injuries unit in the north east of England over a 5-year period. AB - STUDY DESIGN: Retrospective study of the management of patients with a confirmed diagnosis of post-traumatic syringomyelia (PTS) in a regional spinal injuries unit. OBJECTIVES: To investigate the incidence of PTS in a regional spinal injuries unit, and to assess mode of presentation, management, and subsequent outcome. SUMMARY OF BACKGROUND DATA: Majority of patients male, mean age 44 years (range 33-60 years), with thoracic spinal cord injury (SCI). Most PTS developed within 5 years after injury (range 6 months to 25 years) and presented with reduced sensation. Fifty percent had surgical intervention at the time of SCI, and 50% managed conservatively. Four patients had additional spinal injury pathology not corrected at time of surgery. METHODS: Retrospective analysis of the case notes of all patients with a confirmed diagnosis of PTS (n = 16). Demographic details obtained and details of the original injury and subsequent clinical course and management noted. RESULTS: Incidence of PTS = 0.02%. Magnetic resonance imaging scanning performed in 56% with PTS. The majority of PTS developed around the site of the original lesion. The most common method of management was insertion of a syringoperitoneal shunt (44%). Thirty-one percent improved after surgery, 31% remain stable. One patient died. The symptoms of 3 patients continue to deteriorate slowly despite surgical intervention. CONCLUSIONS: Incidence of PTS are lower in our study than that quoted in the literature. Benefits of initial surgical management of SCI in reducing development of PTS are unclear. Benefits of surgical management of PTS are unclear. Additional studies are required as this may influence future management of spinal cord injured patients. PMID- 15897838 TI - Low back pain, work absenteeism, chronic back disorders, and clinical findings in the low back as predictors of hospitalization due to low back disorders: a 28 year follow-up of industrial employees. AB - STUDY DESIGN: Prospective cohort study. OBJECTIVE: To study symptoms, chronic disorders, and clinical findings in the low back, and work absenteeism, as predictors of hospitalization. SUMMARY OF BACKGROUND DATA: Socioeconomic and lifestyle factors are associated with back-related hospitalization, but the significance in the working normal population of low back symptoms and clinical findings are not known. METHODS: The cohort (n = 902) was drawn in 1973 from among employees in the metal industry (n = 2,653). The data were collected by questionnaire and a structured clinical assessment by a physiotherapist. Weight was measured. A sum score of local and radiating low back symptoms (frequency during the past year on a 4-point Likert scale) was categorized as no/yes and no/infrequent/frequent. Local and radiating symptoms were considered also separately. The data were linked with those from the Finnish Hospital Discharge Register during 1973 to 2000. Logistic regression and the Cox proportional hazard models were used. RESULTS: As compared with persons without low back pain, those with frequent or radiating low back pain had an increased risk of hospitalization due to low back disorders (hazard rate ratio (HRR), 3.0; 95% confidence interval (CI), 1.4-6.5, and 3.7; 1.8-7.7, respectively) after adjustment for age, gender, and occupational class. Similarly, clinical findings (HRR, 2.4; 95% CI, 1.3-4.7), back-related absenteeism (HRR, 3.3; 95% CI, 1.6-6.7), and chronic low back disorders (HRR, 2.8; 95% CI, 1.5-5.4) predicted hospitalization. The associations persisted when further adjusted for smoking, body mass index, and distress symptoms at baseline. CONCLUSION: Frequent or radiating low back symptoms, chronic low back disorders, back-related work absenteeism, and having clinical findings in the low back predicted inpatient hospital care for low back disorders. PMID- 15897839 TI - A novel back school using a multidisciplinary team approach featuring quantitative functional evaluation and therapeutic exercises for patients with chronic low back pain: the Japanese experience in the general setting. AB - STUDY DESIGN: A prospective cohort study. OBJECTIVES: To introduce a novel back school for the treatment of patients with chronic low back pain (CLBP), and to report its clinical results. SUMMARY OF BACKGROUND DATA: Although back school is one of the treatment methods for patients with CLBP, its efficacy and appropriate style remain controversial. No studies have been published regarding the combined program of back school with a multidisciplinary team approach. METHODS: A total of 182 patients with CLBP (74 men and 108 women; average age, 43.8 years) participated in this study. All patients were enrolled in the back school using a multidisciplinary team approach featuring quantitative functional evaluation and therapeutic exercises. The following outcome measures were evaluated at the baseline, and 6 and 12 months after the enrollment: the level of pain evaluated with a Visual Analog Scale (VAS), flexibility of trunk and hamstrings (finger floor distance, straight leg raising), trunk muscle strength and endurance, frequency of therapeutic exercises, and self-reported patient satisfaction. RESULTS: An averaged VAS score was 6.2 before enrollment in the program and 2.8 at follow-up. The pain improved in 141 patients (80.8%), did not change in 27 (15.4%), and was aggravated in 7 (3.8%). There was statistically significant improvement of finger-floor distance, trunk muscle strength, and endurance in the patients whose pain was relieved after enrollment in the program (P < 0.05). The compliance with the exercise program was significantly correlated with the clinical results (P < 0.05). CONCLUSIONS: We developed a novel back school using a multidisciplinary team approach, featuring quantitative functional evaluation and therapeutic exercises. The current study demonstrated that our program could provide a satisfactory result for the treatment of patients with CLBP. The quantitative functional evaluation was a worthwhile outcome measure when evaluating the efficacy of the treatment program. Teaching body mechanics and performing the therapeutic exercises through the multidisciplinary team approach are essential to managing CLBP in a general setting. PMID- 15897840 TI - Thoracic microendoscopic discectomy: a human cadaver study. AB - STUDY DESIGN: Feasibility analysis of percutaneous posterolateral thoracic microendoscopic discectomy in a human cadaver model. OBJECTIVE: To describe a new, minimally invasive, posterolateral approach to the thoracic spine for the treatment of disc herniations. SUMMARY OF BACKGROUND: Thoracoscopic discectomy offers surgeons direct ventral access to thoracic disc herniations but requires entry into the chest. Many surgeons favor a posterolateral approach to the thoracic spine, thereby avoiding morbidity associated with entry into the thoracic cavity. By adapting minimal access surgical techniques to the thoracic spine, effective treatment of thoracic disc herniations should be possible and may help expedite recovery. METHODS: Two cadaveric human torsos were used. Using simple adaptations of our standard lumbar microendoscopic discectomy technique, endoscopic discectomies were performed throughout the mid and lower thoracic spine. Operative time was recorded. The extent of the discectomy as well as the extent of bony removal was evaluated using computed tomography myelography. RESULTS: Nine discectomies were performed in two cadaveric specimens, from T5-T6 T9-T10. Operative times ranged from 46 to 77 minutes (mean 60 minutes). The procedure required removing 3.4 mm (+/-1.9 mm) of the ipsilateral facet, which amounted to 35.4% (+/-17.5%) of the facet complex. Canal decompression averaged 73.5% (+/-7.9%). CONCLUSIONS: Thoracic microendoscopic discectomy allows for a posterolateral approach to thoracic disc herniation without entry into the chest cavity that consistently gives access to the majority of the canal while requiring only a minimal amount of bone removal. This technique provides an approach angle similar to that obtained with other posterolateral discectomy techniques while limiting the morbidity associated with exposure. PMID- 15897841 TI - The prognostic value of functional capacity evaluation in patients with chronic low back pain: part 1: timely return to work. And part 2: sustained recovery. PMID- 15897843 TI - A simple technique to facilitate the Le Fort I osteotomy downfracture. AB - In order to mobilize and reposition the maxilla when treating dentofacial deformities, the Le Fort I maxillary osteotomy is commonly used. Like other surgical procedures, this procedure is not without inherent difficulties. At times, it may be difficult to initially mobilize the maxilla after performing the osteotomies. Techniques used to help in the initial mobilization and downfracture of the maxilla from the pterygoid plates include digital pressure to the dentoalveolus, disimpaction forceps, spreaders, and other instruments. These techniques are similar in that they all apply a unidirectional downward vector of force on the maxilla and have various drawbacks. We present a technique that uses a 0.5-mm stainless steel traction wire to facilitate Le Fort I maxillary downfracturing by applying a forward and downward vector of force simultaneously. After vertical downfracture of the maxilla, lateral traction on the wire can be used to complete the pterygomaxillary disjunction. This technique has several advantages and is atraumatic, and requires no special instrumentation; thus, downfracturing of the maxilla is done with relatively little effort and minimal risk. PMID- 15897844 TI - Enteric bacteria mandibular osteomyelitis. AB - Osteomyelitis of the mandible is a relatively rare inflammatory disease that usually stems from the odontogenic polymicrobial flora of the oral cavity. We are reporting 2 unusual cases of mandibular osteomyelitis resulting from enteric bacteria infection. In one patient, abundant clinical evidence suggested a diagnosis of a chronic factitious disease, whereas in the second patient no obvious etiology was found. PMID- 15897845 TI - Efficacy of preoperative oral rofecoxib in pain control for third molar surgery. AB - OBJECTIVE: The study compared the analgesic efficacy of preoperative 50 mg oral rofecoxib, 400 mg ibuprofen, and placebo in the control of postoperative pain after third molar surgery. STUDY DESIGN: This was a clinical randomized, double blind, cross-over, placebo- and active-comparator-controlled study. The surgeries were randomized into 3 groups, in which patients were given a single dose of 50 mg rofecoxib, 400 mg ibuprofen, or placebo 30-60 minutes before the surgery. The patients were asked to quantify their pain intensity basing on a visual analog scale postoperatively. A rescue medication, 500 mg acetaminophen, was prescribed to the patients. The quantity and time of consumption of the rescue tablets were recorded by the patients. RESULTS: A total of 49 patients completed the study. Of the 98 lower third molar extractions 33 were with rofecoxib, 33 ibuprofen, and 32 placebo. The pain scores within the first 6 hours postoperatively in the rofecoxib group were significantly lower than the placebo ( P < .05). The ibuprofen group did not have significantly lower pain scores than the placebo group. Regarding the postoperative requirement of rescue medication, the rofecoxib group required significantly less rescue medication than both ibuprofen and placebo groups in the first twelve hours after the surgery. CONCLUSIONS: The preoperative oral intake of 50 mg rofecoxib provides a significantly better analgesic benefit than the placebo for postoperative pain relief in the first 6 hours after third molar surgery. This regimen also reduced the requirement of postoperative analgesic when compared with ibuprofen and placebo in the first 12 postoperative hours. PMID- 15897846 TI - Modified cheek retractor. PMID- 15897847 TI - Time to rethink publish or perish. PMID- 15897848 TI - Cytogenetic analysis of oral malignant melanoma. PMID- 15897849 TI - General residency programs after graduation. PMID- 15897850 TI - Neurosensory changes in the infraorbital nerve following zygomatic fractures. AB - OBJECTIVE: To document the neurosensory changes in the infraorbital nerve following zygomatic fractures managed in various ways. STUDY DESIGN: Twenty-five patients were included in the study. Neurosensory function was assessed with calibrated nylon monofilaments, electrical stimulation, heat detection thresholds and response to pin prick in the infraorbital, supraorbital, and mental nerve regions. Patients were seen immediately post-trauma, then 1 and 6 months following surgery. RESULTS: Nine fractures were caused by traffic accidents (TAs), 8 by falls, and 8 by a local blow in a physical dispute. The fractures consisted of 15 displaced and 10 minimally or nondisplaced zygomatic complex fractures, and were left surgically untreated in 7 cases (None group), reduced but not fixed in 8 cases (Reduction group), and fixed with plates in 10 cases (Plates group). Plates were employed significantly more often in displaced fractures (chi-squared P = .0006). At 6 months significantly improved infraorbital nerve function was found in the Plate and None groups relative to the Reduction group (ANOVA P = .006). Only 1 case of chronic neuropathic pain was found. CONCLUSIONS: This study concurs with previous studies in finding that plate fixation allows for significantly better restoration of infraorbital nerve function. Chronic neuropathic pain following zygomatic fractures is rare. PMID- 15897851 TI - Molecular correlates of temporomandibular joint disease. AB - OBJECTIVE: The relation between disease severity and the known mediators of pain, inflammation, and tissue damage-prostaglandin E 2 (PGE 2 ), leukotriene B 4 (LTB 4 ), malondialdehyde (MDA), nitric oxide (NO), and myeloperoxidase (MPO)-was examined in the synovial fluid of patients with internal derangement (ID) of the temporomandibular joint (TMJ). STUDY DESIGN: Thirty-two patients with ID were classified according to Wilkes by clinical and radiological examinations, and TMJ synovial fluid samples were obtained by arthrocentesis. PGE 2 and LTB 4 levels were measured by ELISA kits, MDA levels were determined by a fluorometric method, myeloperoxidase activity was determined by an end-point method, and NO levels were measured by Griess reaction. RESULTS: The earliest significant increase was observed in NO levels (stage II) and this elevation persisted in the subsequent stages. The first significant elevation in PGE 2 and LTB 4 levels and MPO activity were observed in stage III. Both PGE 2 and LTB 4 levels were increased in stage III and were correlated with each at this stage and in the subsequent stage. Significant increases in MDA levels were observed only in stage IV. At stage IV there was correlation between MDA and PGE 2 , MDA and LTB 4 , and MDA and MPO. The relation between PGE 2 and MDA was the most powerful one. CONCLUSION: Results of this cross-sectional study point out the relation between disease severity and levels of some molecular mediators in synovial fluid of TMJ. Longitudinal studies are needed to explore the role of these molecular mediators in the progression of ID. PMID- 15897852 TI - Complications of free radial forearm flap transfers for head and neck reconstruction. AB - Free tissue transfer using microvascular anastomosis has been established as an accepted maxillofacial reconstructive procedure. The free radial forearm flap (FRFF) has become a workhorse flap as a means of reconstructing surgical defects in the head and neck region. Since 1992, we have carried out 38 FRFF transfers in 37 patients for reconstruction after head and neck cancer ablative surgery. We present our clinical experience with head and neck reconstruction using the FRFF and the morbidity of the donor sites. Of the 38 FRFFs, 35 FRFFs were performed successfully. The survival rate of FRFF was 92%. Donor site complications included partial loss of skin graft in 4 donor sites (11%), abnormal sensations in 10 (26%), poor appearance in 3 (8%), and reduced grip strength in 4 (11%). Therefore, we believe that, because of the reliability, functional characteristics, and low donor site morbidity, the FRFF is a useful and versatile flap for reconstruction of head and neck defects. PMID- 15897853 TI - Statistical correlation between pharyngitis and temporomandibular joint disease. AB - OBJECTIVE: The objective of this study was to evaluate the upper respiratory infection (URI) as an aggravating factor in the established temporomandibular joint (TMJ) disease. PATIENTS AND METHODS: Four hundred seventeen patients suffering from temporomandibular disorder (TMD) were selected and investigated by means of questionnaires and clinical examinations. After excluding the patients with only muscle disorders, 283 patients were included for the association study between TMJ disease and infectious conditions. The screened infectious conditions were otitis media, maxillary sinusitis/rhinitis, and pharyngitis/tonsillitis. The chi-square test was used to determine the association between variables and stepwise logistical regression was then used. RESULTS: The prevalence of maxillary sinusitis/rhinitis in TMD patients was 7.0%. The patients who had mouth opening limitation were 9.93 times more likely to have maxillary sinusitis/rhinitis than those without ( P = .0004). The prevalence of tonsillitis/pharyngitis in TMD patients was 9.1%. The patients who had mouth opening limitation were 3.50 times more likely to have tonsillitis/pharyngitis than those without it ( P = .0028). The patients who had TMJ capsulitis were 3.91 times more likely to have tonsillitis/pharyngitis than those without it ( P = .0028). CONCLUSION: The conclusion is made that pharyngitis/sinusitis is significantly associated with some clinical symptoms of TMD. The infection of closely related anatomical structures with TMJ may have an influence on TMJ symptoms. Thus, pharyngitis/sinusitis in the established TMD patients can be a significant warning sign for TMJ symptoms to appear shortly thereafter. PMID- 15897854 TI - Post-traumatic bifid condyle associated with temporomandibular joint ankylosis: report of a case and review of the literature. AB - Bifid condyle is a rare condition. Most initially reported cases were found in studies conducted on skeletal specimens. While increasing numbers are being reported on living persons, most of them are asymptomatic and have been found on routine dental radiographic examination for other dental complaints. Most of the cases of bifid condyle reported so far have occurred unilaterally and predominantly on the left side. Bifid condyle associated with temporomandibular joint ankylosis is very rare with only 2 cases reported in the English-language literature as far as we know. An additional case of bifid condyle associated with temporomandibular joint ankylosis, involving the right side of mandible, is presented as well as a review of the literature on bifid condyles including those associated with temporomandibular joint ankylosis. PMID- 15897855 TI - Oral and dental manifestations in diabetic and nondiabetic uremic patients receiving hemodialysis. AB - OBJECTIVE: The aim of this study was to examine the dental condition and oral manifestations in diabetic and nondiabetic uremic patients undergoing hemodialysis. STUDY DESIGN: A total of 128 patients undergoing hemodialysis therapy were classified into the diabetic and nondiabetic groups and examined for uremic oral manifestations, dental caries, and the periodontal status. All the patients received predialytic salivary pH examination. In the diabetic group, the correlation between oral findings and glycemic controlled levels, which was collected based on Hb A1C values, were further studied. RESULTS: The diabetic group exhibited significantly higher prevalence of caries and more severe dry mouth, taste change, and mucosa pain than the nondiabetic group. The diabetic group tended to have lower predialytic salivary pH, and patients with poor glycemic control (ie, Hb A1C > 9%) showed higher incidence of dry mouth, mucosal pain, and tongue coating. However, the DMFT and CPI index were not associated with glycemic control in the diabetic group. CONCLUSIONS: This study reveals that diabetic uremic patients undergoing maintained hemodialysis exhibited a potentially higher risk for dental decay and xerostomia. Lower salivary pH and poor glycemic control may affect oral manifestations. Further research is needed to clarify the combined influence of diabetic nephropathy on oral health. PMID- 15897856 TI - Scoring system for monitoring oral lichenoid lesions: a preliminary study. AB - BACKGROUND: Currently, there is no universal system for scoring the severity of lichenoid lesions of the oral cavity that is easy to use, reproducible and that is representative of the different clinical forms of the disease. OBJECTIVES: In this study, a scoring system was developed to monitor the severity of oral lichen planus (OLP) and chronic oral graft-versus-host disease (cGVHD). METHODS: Six patients with OLP and three with cGVHD were studied. This scoring system was utilized to compare the severity of disease within and between patients. Three investigators independently reviewed photographs taken from each patient and scored the severity of disease. Inter and intra-observer consistency was evaluated using the Spearman rank correlation analyses. RESULTS: Intra-observer reliability was 0.98 (Spearman correlation coefficient) and inter-observer reliability was 1.00. CONCLUSION: This scoring system is easy-to-use, reproducible, representative of the severity of the disease and useful for monitoring OLP and cGVHD. PMID- 15897857 TI - Impact of dry mouth on oral symptoms and function in removable denture wearers in Japan. AB - OBJECTIVE: The purpose of this study was to determine the association of dry mouth with oral symptoms and function, such as denture instability, discomfort, soreness in denture-bearing tissue, and dissatisfaction with chewing, tasting, or speaking in removable denture wearers. STUDY DESIGN: The subjects were 493 removable denture wearers with a mean age of 67.3 years. Perception of oral dryness was measured by a questionnaire. A multiple logistic regression analysis was used to determine the effect of the dry mouth on oral symptoms and function after controlling for age and gender. RESULTS: Oral dryness during eating was related to dissatisfaction with chewing (odds ratio, 10.5; P < .001) and speaking (odds ratio, 3.5; P < .05) and overall dissatisfaction (odds ratio, 6.3; P < .01) in complete denture wearers. Feeling of dry mouth was likely to be associated with soreness in denture-bearing tissues in both complete and removable partial denture wearers. CONCLUSIONS: There was a significant association of the perception of dry mouth among a group of denture wearers with oral symptoms and function. PMID- 15897858 TI - The relationship between PTC taster status and taste thresholds in young adults. AB - OBJECTIVE: The aim of this study was to compare taste detection and recognition thresholds of young males and females, and young phenylthiocarbamide (PTC) "tasters" and "nontasters" for stimuli representing sweet, sour, bitter, salty, and umami classes of taste sensations. STUDY DESIGN: Thirty-eight men and 37 women (mean [SD] age = 24.5 [2.5] years) were classified as PTC tasters and nontasters according to their PTC recognition thresholds. Detection and recognition thresholds for the non-PTC stimuli were determined using a 2 alternative, forced choice procedure. RESULTS: The detection thresholds for quinine-HCl and sucrose and the recognition threshold for quinine-HCl were significantly higher in the PTC nontasters than in the tasters. The PTC threshold showed significant correlations with detection or recognition thresholds for sucrose, sodium chloride, quinine-HCl, and monosodium glutamate. The sucrose recognition threshold was lower in women than in men. CONCLUSION: In this study, gender and PTC taster status were found to be associated with thresholds for sucrose and quinine-HCl. PMID- 15897859 TI - Oral inflammatory myofibroblastic tumor demonstrating ALK, p53, MDM2, CDK4, pRb, and Ki-67 immunoreactivity in an elderly patient. AB - Inflammatory myofibroblastic tumor (IMT) is a controversial lesion composed of myofibroblasts, accompanied by varying numbers of inflammatory cells. Various pathogenetic factors have been proposed (ie, reactive, infectious, autoimmune, and neoplastic) but the etiology of most IMTs remains unknown. Here we review the literature of oral IMTs, detailing the demographic profile of these rare lesions. Moreover, we present an unusual case of IMT arising from the mandibular alveolar mucosa of an 82-year-old female. Microscopic examination revealed plump spindle cells set in a myxoid vascular stroma admixed with inflammatory cells. Numerous large ganglion cell-like cells were seen, some exhibiting emperipolesis of neutrophils. Ultrastructurally, prominent myofibroblasts with abundant rough endoplasmic reticulum were noted. Tumor cells were immunoreactive for vimentin, smooth muscle actin, and KP1 (CD68), and negative for desmin, S-100, and EBV-LMP. The lesion was excised without margins and the patient has manifested no evidence of disease at an 18-month recall. In an attempt to further delineate the potential neoplastic nature of this lesion, we assessed the immunohistochemical expression of various markers that have been linked to neoplastic transformation. The recorded positivity for ALK, p53, MDM2, CDK4, pRb, and Ki-67, despite the absence of bcl-2 reactivity, strongly favors the neoplastic origin of the studied tumor. PMID- 15897860 TI - Paget's disease of bone in a Chinese patient: a case report and review of the literature. AB - Although it is fairly common in the Western countries, Paget's disease of bone is very rare in Chinese individuals. Through an extensive literature search, only 8 Chinese Paget's disease cases were found in the English language literature and all were reported in local medical journals of Asia that may not be accessible to international readers. To enhance the awareness of the rarity of Paget's disease in Chinese individuals to pathologists worldwide, we present a case of Paget's disease in a 54-year-old Chinese male. We also compare the clinical features of Paget's disease reported in Chinese patients to Paget's disease reported in the Western countries. No familial cases and no malignant transformation are found in the reported cases of Paget's disease in Chinese patients. In addition, more often skull involvement, higher frequency of monostotic cases, and symptomatic cases are observed in this limited number of Paget's disease cases reported in Chinese individuals. PMID- 15897861 TI - Magnetic resonance images of the temporomandibular joints of patients with acquired open bite. AB - OBJECTIVES: Acquired anterior open bites were reported as the consequence of condylar collapse, which was associated with inflammatory TMJ disorders. However, we have seen such malocclusion patients whose condylar changes seemed to be related to TMJ degeneration associated with internal derangement. The aims of this study were to review the clinical history and to study the TMJ MRI of these patients. STUDY DESIGN: TMJ MRIs of patients, who had presented acquired anterior open bite at first visit, were retrieved from the image database for the analysis. Clinical histories focused on internal derangement were collected retrospectively. The soft tissue and hard tissue changes disclosed by MRI were also studied. RESULTS: All patients had experienced common signs/symptoms of TMJ internal derangement. All affected TMJs had anteriorly displaced disks and degenerative changes. Horizontally destructed condylar forms were seen significantly more frequently in these patients. CONCLUSION: TMJ degeneration associated with displaced disks might be a cause leading to the development of acquired anterior open bite. PMID- 15897862 TI - Dosimetric evaluation of the effect of dental implants in head and neck radiotherapy. AB - OBJECTIVE: The aim of the study was to examine the dose enhancement from scattered radiation at bone-dental implant interfaces during simulated head and neck radiotherapy. STUDY DESIGN: Four cylindrical titanium dental implants with 3 different sizes and lengths were implanted into a human mandible in 4 different positions. Ionization measurements for 6 MV X, 25 MV X, and Co-60 gamma rays were done. Thermoluminescent dosimeter (TLD 100 ) chips were used to measure radiation dose enhancement due to the scattered electrons from titanium and electronic disequilibrium at the tissue-metal interface. RESULTS: The results showed that for Co-60, there is a 21% maximum increase in dose to alveolar mandibular bone at the close proximity to the titanium. For 6-MV x-rays the dose enhancement increase was almost the same or slightly lower than for Co-60, while for 25-MV high-energy x-rays, dose enhancement was lower than that of others. This increase in dose enhancement fell off rapidly and became insignificant at 2 mm from the interface. CONCLUSION: Total dose that may lead to osteoradionecrosis risk of the mandible is slightly but not significantly affected by the scattered dose of the dental implants of lower jaw in the radiation field exposed to 3 different radiation beams. PMID- 15897863 TI - Comparison of linear tomography and direct ridge mapping for the determination of edentulous ridge dimensions in human cadavers. AB - OBJECTIVE: The purpose of this study was to compare the accuracy of linear tomography (LT) and direct ridge mapping (RM) for determining alveolar ridge dimensions. STUDY DESIGN: One site in the posterior mandible was selected for evaluation in each of 5 cadaver heads. Vacuum-formed stents made from models of the cadaver ridges were used to identify 3 sets of measurement points for each specimen: coronal (intersection of coronal and middle third of ridge), middle (intersection of middle and apical third), and apical (base of vestibule). The imaging stent contained 2-mm metal balls at each point, while the RM stent had holes drilled at corresponding locations. Linear tomograms and periapical radiographs (PA) were taken of the selected sites. RM measurements were made with calipers. Five blinded examiners measured ridge width at the designated measurement points with both LT and RM as well as distance from the ridge crest to mandibular canal (using PAs for RM group). Mandibles were then sectioned and an independent examiner made direct measurements (DM). MANOVA was used to determine whether LT and RM differed significantly from DM. RESULTS: There were no significant differences between LT and RM for ridge width measurements. However, both techniques underestimated ridge dimensions compared to DM ( P < .05). Measurement of mandibular canal height was accurate when determined by periapical radiographs but not by LT. CONCLUSION: Neither LT nor RM proved to be completely accurate in determining ridge width in the posterior mandible. PMID- 15897864 TI - An investigation of microleakage from root-end fillings in ultrasonic retrograde cavities with or without finishing: a quantitative analysis. AB - OBJECTIVE: The aim of this study was to make a quantitative assessment of the sealing ability of Super-EBA, IRM, and Pro Root MTA root-end fillings subjected to 3 different finishing techniques. STUDY DESIGN: Eighty-one ultrasonically prepared root-end cavities in human canines were separated randomly into 3 test groups of 27 roots each. The cavities were filled with Super-EBA, IRM, or Pro Root MTA and finished by ball burnishing. Sequentially, 18 roots from each group received a final smoothing with either a 30-fluted tungsten carbide finishing bur or a Zekrya carbide 28-mm bur. Samples were prepared and immersed in 2% methylene blue dye neutral solution for 12 hours. Roots were ground into a powder and prepared for analysis in an absorbency spectrophotometer. RESULTS: The results revealed that Pro Root MTA displayed significantly less mean dye microleakage ( P < .05) than Super-EBA and IRM root-end fillings. The Super-EBA root-end fillings, although presenting a greater mean dye microleakage, did not differ significantly from IRM ( P > .05). The finishing technique did not significantly ( P > .05) affect the incidence of microleakage among the materials tested. CONCLUSIONS: The favorable results obtained with MTA in leakage studies may be related to its good marginal adaptation. Spectrophotometric analysis may provide valuable information about the sealing capacity of root-end fillings. None of the procedures tested were able to avoid leakage, a finding that stresses the importance of the eradication of irritants within the root canal system. PMID- 15897865 TI - Enamel matrix derivative inhibits TNF-alpha-induced apoptosis in osteoblastic MC3T3-E1 cells. AB - OBJECTIVE: The purpose of this study was to examine the effect of enamel matrix derivative (EMD) on TNF-alpha-induced apoptosis in osteoblastic MC3T3-E1 cells. STUDY DESIGN: MC3T3-E1 cells were cultured at an initial density of 5000/cm 2 in Dulbecco's modified eagle medium (DMEM) with 10% fetal bovine serum (FBS) and allowed to adhere for 24 hours. Medium was then changed into DMEM with 0.5% FBS. After 16 hours, cells were treated with EMD (100 microg/mL) alone, tumor necrosis factor alpha (TNF-alpha) (20 ng/mL) alone, transforming growth factor beta 1 (TGF beta1) (10 ng/mL) alone, TNF-alpha plus TGF-beta1, or TNF-alpha plus EMD. Cells cultured with DMEM and 0.5% FBS served as control. Following 24-hour incubation, apoptosis was assessed by terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) assay, and quantified by cell death enzyme-linked immunosorbent assay (ELISA). RESULTS: Both TUNEL assay and cell death ELISA show that TNF-alpha induces apoptosis in MC3T3-E1 cells. TNF-alpha increases cell death by approximately 2-fold, which is attenuated by both EMD and TGF-beta1. CONCLUSION: Like TGF-beta1, EMD protects osteoblasts from inflammation-induced apoptosis. PMID- 15897866 TI - In vitro assessment of the immediate and prolonged antimicrobial action of chlorhexidine gel as an endodontic irrigant against Enterococcus faecalis. AB - OBJECTIVE: The aim of this study was to assess in vitro the antimicrobial activity of 2% chlorhexidine gel against Enterococcus faecalis , comparing it to other endodontic irrigants (2% chlorhexidine liquid and 5.25% sodium hypochlorite). STUDY DESIGN: Eighty roots of human mandibular premolars were prepared by serial instrumentation technique, autoclaved, and contaminated for 7 days with E faecalis monocultures. The roots were then divided into 5 groups according to the irrigant substance used during the standardized biomechanical preparation. In order to evaluate the antimicrobial action of the irrigant substances, 3 microbial samples were taken: initial (before the biomechanical preparation); post-treatment (immediately after the biomechanical preparation), and final (7 days after the biomechanical preparation). The microbiological samples were plated to count the colony-forming units (CFU). RESULTS: The 2% chlorhexidine gel and 2% chlorhexidine liquid significantly reduced the E faecalis CFU in the post-treatment and final microbiological samples. The 5.25% sodium hypochlorite also reduced the E faecalis CFU immediately after the root canal instrumentation, but it was not able to keep the root canal free of detectable E faecalis in the final sample. CONCLUSIONS: The 2% chlorhexidine gluconate (gel and liquid) antimicrobial ability was more effective than 5.25% sodium hypochlorite in keeping low CFU of E faecalis for 7 days after the biomechanical preparation. PMID- 15897867 TI - Oligomerization of Evi-1 regulated by the PR domain contributes to recruitment of corepressor CtBP. AB - Evi-1 is a transcription factor that is implicated in leukemic transformation of hematopoietic cells. Two distinct alternative forms, Evi-1a and Evi-1c, are generated from the EVI-1 gene. Whereas Evi-1a is widely recognized as an oncoprotein, a role for Evi-1c, which has an additional PR domain in the amino terminus of Evi-1a, in leukemogenesis, has not been elucidated thus far. Aberrant oligomerization of transcription factors has recently emerged as a prevalent mechanism for activating their oncogenic potential in hematopoietic malignancies. Here, to study the mechanisms that underlie Evi-1-mediated oncogenesis, we investigated formation of oligomeric complexes by the Evi-1 proteins. We show that Evi-1a forms homo-oligomers, whereas Evi-1c exclusively exists as a monomer in mammalian cells. Remarkably, Evi-1c has lost the ability to interact with CtBP, a transcriptional corepressor that associates with Evi-1a. As a consequence, the ability of Evi-1c to repress transforming growth factor-beta (TGF-beta) signaling is significantly abrogated. These results identify a novel function of a PR domain to regulate oligomerization of transcription factors and suggest that homo-oligomerization may play a critical role in corepressor recruitment by the Evi-1 proteins. In addition, we found that the chimeric oncoprotein acute myelocytic leukemia (AML)1-Evi-1, generated in t(3;21) leukemia, also forms homo-oligomers and hetero-oligomers with Evi-1a, while it did not interact with Evi-1c. Consistent with the results, repression of TGF-beta by AML1-Evi-1 was significantly enhanced by Evi-1a, whereas it was hardly affected by the presence of Evi-1c. These results suggest that oligomerization may contribute to the oncogenic potential of Evi-1-containing proteins. PMID- 15897868 TI - Array-based comparative gene expression analysis of tumor cells with increased apoptosis resistance after hypoxic selection. AB - Tumor hypoxia is an adverse prognostic factor. In a recent study, we could demonstrate that cyclic hypoxia selects for hypoxia-tolerant tumor cells, which are cross-resistant to other stimuli of mitochondrial death pathways. In contrast, sensitivity of the cells to death-receptor ligands was mainly not affected. The aim of the present study was to further elucidate cellular changes induced by cyclic hypoxia and to identify alterations in gene expression pattern upon hypoxic selection by means of DNA-microarray analysis. Our data reveal that cyclic hypoxia resulted in the selection of cells with resistance to doxorubicine and radiation. Furthermore, hypoxic selection was accompanied by constitutive changes of the gene expression pattern with downregulation of 156 and upregulation of 82 genes. Most of the differentially regulated genes were involved in cellular responses to hypoxia and reoxygenation. While many of the genes that were downregulated upon hypoxic selection represent genes that are usually upregulated by acute hypoxia, the genes that were upregulated represent genes that are involved in stress resistance and anti-apoptotic signalling. Most importantly, hypoxic selection was not associated with changes of single apoptosis relevant genes, but with alterations in gene expression levels of a wide variety of genes indicating a more complex adaptation process. PMID- 15897869 TI - In silico whole-genome scanning of cancer-associated nonsynonymous SNPs and molecular characterization of a dynein light chain tumour variant. AB - Last decade has led to the accumulation of large amounts of data on cancer genetics, opening an unprecedented access to the mapping of cancer genes in the human genome. Single-nucleotide polymorphisms (SNPs), the most common form of DNA variation in humans, emerge as an invaluable tool for cancer association studies. These genotypic markers can be used to assay how alleles of candidate genes correlate with the malignant phenotype, and may provide new clues into the genetic modifications that characterize cancer onset. In this cancer-oriented study, we detail an SNP mining strategy based on the analysis of expressed sequence tags among publicly available databases. Our whole-genome approach provides a comprehensive and unbiased description of nonsynonymous SNPs (nsSNPs) in tumoral versus normal tissues. To gain further insights into the possible relationships between genetic variation and altered phenotype, locations of a subset of nsSNPs were mapped onto protein domains known to be critical for protein function. Computational methods were also used to predict the potential impact of these cancer-associated nsSNPs on protein structure and function. We illustrate our approach through the detailed biochemical and structural characterization of a previously unknown cancer-associated mutation (G79C) affecting the 8 kDa dynein light chain (DNCL1). PMID- 15897870 TI - Human and mouse mesotheliomas exhibit elevated AKT/PKB activity, which can be targeted pharmacologically to inhibit tumor cell growth. AB - Malignant mesotheliomas (MMs) are very aggressive tumors that respond poorly to standard chemotherapeutic approaches. The phosphatidylinositol 3-kinase (PI3K)/AKT pathway has been implicated in tumor aggressiveness, in part by mediating cell survival and reducing sensitivity to chemotherapy. Using antibodies recognizing the phosphorylated/activated form of AKT kinases, we observed elevated phospho-AKT staining in 17 of 26 (65%) human MM specimens. In addition, AKT phosphorylation was consistently observed in MMs arising in asbestos-treated mice and in MM cell xenografts. Consistent with reports implicating hepatocyte growth factor (HGF)/Met receptor signaling in MM, all 14 human and murine MM cell lines had HGF-inducible AKT activity. One of nine human MM cell lines had elevated AKT activity under serum-starvation conditions, which was associated with a homozygous deletion of PTEN, the first reported in MM. Treatment of this cell line with the mTOR inhibitor rapamycin resulted in growth arrest in G1 phase. Treatment of MM cells with the PI3K inhibitor LY294002 in combination with cisplatin had greater efficacy in inhibiting cell proliferation and inducing apoptosis than either agent alone. Collectively, these data indicate that MMs frequently express elevated AKT activity, which may be targeted pharmacologically to enhance chemotherapeutic efficacy. These findings also suggest that mouse models of MM may be useful for future preclinical studies of pharmaceuticals targeting the PI3K/AKT pathway. PMID- 15897871 TI - Methylseleninic acid sensitizes prostate cancer cells to TRAIL-mediated apoptosis. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a cytotoxic agent that preferentially induces apoptosis in a variety of human cancer cells. Unfortunately, some tumor cells remain resistant to TRAIL. Therefore, agents that sensitize malignant cells to TRAIL-mediated cell death might be of particular importance for the development of novel antitumor therapeutic regimens. Recent studies establish a critical role of selenium in prostate cancer prevention in vitro and in vivo. Here, we demonstrate that concomitant administration of TRAIL and methylseleninic acid (MSA) produces synergistic effects on the induction of apoptosis in androgen-dependent LNCaP and androgen-independent DU-145 prostate cancer cells. MSA rapidly and specifically downregulates expression of the cellular FLICE inhibitory protein, a negative regulator of death receptor signaling. In addition, we demonstrate that the synergistic effects of MSA and TRAIL result from the activation of the mitochondrial pathway-mediated amplification loop. Addition of MSA effectively blocked TRAIL-mediated BAD phosphorylation at Ser112 and Ser136 in DU-145 cells and was accompanied by induction of the mitochondrial permeability transition and release of apoptogenic cytochrome c and Smac/DIABLO proteins from the mitochondria and into the cytosol. These results suggest that selenium-based dietary compounds may help to overcome resistance to TRAIL-mediated apoptosis in prostate cancer cells. PMID- 15897872 TI - A 1 Mb minimal amplicon at 8p11-12 in breast cancer identifies new candidate oncogenes. AB - Amplification of 8p11-12 is a well-known alteration in human breast cancers but the driving oncogene has not been identified. We have developed a high-resolution comparative genomic hybridization array covering 8p11-12 and analysed 33 primary breast tumors, 20 primary ovarian tumors and 27 breast cancer cell lines. Expression analysis of the genes in the region was carried out by using real-time quantitative PCR and/or oligo-microarray profiling. In all, 24% (8/33) of the breast tumors, 5% (1/20) of the ovary tumors and 15% (4/27) of the cell lines showed 8p11-12 amplification. We identified a 1 Mb segment of common amplification that excludes previously proposed candidate genes. Some of the amplified genes did not show overexpression, whereas for others, overexpression was not specifically attributable to amplification. The genes FLJ14299, C8orf2, BRF2 and RAB11FIP, map within the 8p11-12 minimal amplicon, two have a putative function consistent with an oncogenic role, these four genes showed a strong correlation between amplification and overexpression and are therefore the best candidate driver oncogenes at 8p12. PMID- 15897873 TI - Muc1 affects c-Src signaling in PyV MT-induced mammary tumorigenesis. AB - MUC1 is an integral membrane mucin glycoprotein that is normally expressed on the apical surface of most simple, secretory epithelia and hematopoietic cells. Overexpression of aberrantly glycosylated MUC1 is a hallmark of many carcinomas including 90% of breast carcinomas. MUC1 has been shown to bind to c-Src tyrosine kinase in vitro, whereby c-Src phosphorylates the MUC1 cytoplasmic domain at a YEKV motif. c-Src is an extensively studied nonreceptor tyrosine kinase implicated in mammary tumorigenesis. Previously, mouse mammary tumor virus-driven polyoma middle T-antigen (MMTV-PyV MT) transgenic mice crossed onto a Muc1 null background exhibited a significant delay in tumor progression. c-Src has been shown to interact with PyV MT, and to play an integral and indispensable role in MMTV-PyV MT-induced mammary tumorigenesis. Here, we determine the effect of Muc1 expression on c-Src activation and signaling. Examination of MMTV-PyV MT glands on a wild-type or Muc1 null background demonstrates that Muc1 expression promotes c-Src signaling by influencing its association with known substrates such as the p85 subunit of phosphatidylinositol 3-kinase and beta-catenin. These findings may provide a mechanism for the delay in tumor progression that is observed in the absence of Muc1. PMID- 15897874 TI - Integrin-linked kinase activity regulates Rac- and Cdc42-mediated actin cytoskeleton reorganization via alpha-PIX. AB - Cell spreading and migration are regulated in a Rho family GTPase-dependent manner by growth factors and integrin-mediated cell-extracellular matrix (ECM) interactions. The molecular mechanisms involved in the ECM- and growth factor mediated activation of these small GTPases remain unclear. In the present study, we demonstrate that integrin-linked kinase (ILK), which is a focal adhesion protein activated by both ECM and growth factors, is required for the activation of Rac and Cdc42 in epithelial cells. Ectopic expression of active ILK in mammary epithelial cells induces dramatic reorganization of the actin cytoskeleton and promotes rapid cell spreading on fibronectin. These effects are associated with constitutive activation of both Rac and Cdc42, but not Rho. The use of ILK siRNA or small molecule inhibitors to inhibit ILK expression and kinase activity, respectively, results in diminished cell spreading and actin cytoskeleton reorganization, concomitant with a reduction in Rac and Cdc42 activation. Studies into the mechanism of ILK-mediated Rac activation suggest an important role for the ILK-beta-parvin interaction and the activity of the Rac/Cdc42-specific guanine nucleotide exchange factor alpha-PIX downstream of ILK. Taken together, these data demonstrate an essential role of ILK kinase activity in Rac- and Cdc42 mediated actin cytoskeleton reorganization in epithelial cells, further solidifying a role for ILK in the regulation of cancer cell motility and invasiveness. PMID- 15897875 TI - Paired-like homeoprotein ESXR1 acts as a sequence-specific transcriptional repressor of the human K-ras gene. AB - Gain-of-function mutation of the K-ras gene is one of the most common genetic changes in human tumors. In tumors carrying K-ras mutation, the presence of oncogenic K-Ras is necessary for maintenance of the transformed phenotype. ESXR1 is a human paired-like homeodomain-containing protein expressed primarily in the testis. In cells, the 65-kDa full-length ESXR1 protein is proteolytically processed into an N-terminal 45-kDa fragment containing the homeodomain, which localizes exclusively within the nucleus, and a C-terminal 20-kDa fragment consisting of a proline-rich repeat region, which is located in the cytoplasm. In this work, we demonstrated that the N-terminal ESXR1 fragment specifically recognizes the TAATNNNATTA P3 consensus sequence for the paired-like homeodomain and functions as a sequence-specific transcriptional repressor. We also showed that the N-terminal ESXR1 fragment binds to the TAATGTTATTA sequence present within the first intron of the human K-ras gene and inhibits its expression at both mRNA and protein levels. Ectopic expression of the N-terminal ESXR1 fragment in human carcinoma cells that carry mutated K-ras reduces the level of K-Ras and thereby inhibits the tumor cell proliferation. Identification of ESXR1 as a transcriptional repressor of K-ras has an important implication for the development of cancer therapy that inhibits oncogenic K-Ras expression. PMID- 15897876 TI - CBX7 controls the growth of normal and tumor-derived prostate cells by repressing the Ink4a/Arf locus. AB - Control of cell proliferation by Polycomb group proteins (PcG) is an important facet of cellular homeostasis and its disruption can promote tumorigenesis. We recently described CBX7 as a novel PcG protein controlling the growth of normal cells. In an attempt to identify a putative role of CBX7 in tumorigenesis, we analysed CBX7 expression in a panel of cancer cell lines and primary tissues. CBX7 was highly expressed in three different prostate cancer cell lines and present at elevated levels in normal prostate. Ablation of CBX7 expression using short hairpin RNAs (shRNA) resulted in upregulation of p16Ink4a and p14Arf in both LNCaP and PC-3 prostate cell lines. CBX7 knockdown caused an impairment of cell growth that was dependent on the status of the p14Arf/p53 and p16Ink4a/Rb pathways in both normal and cancer prostate cells. CBX7 overexpression in LNCaP cells resulted in a slight growth advantage in both androgen-dependent and independent conditions. Moreover, CBX7 expression cooperated with c-Myc in rendering LNCaP cells insensitive to growth arrest by androgen receptor inhibition. Together, these data suggest that CBX7 represses p16Ink4a and p14Arf expression in normal and tumor-derived prostate cells, affecting their growth depending on the status of the p16Ink4a/Rb and the p14Arf/p53 pathways. PMID- 15897877 TI - Activation of the neuregulin-1/ErbB signaling pathway promotes the proliferation of neoplastic Schwann cells in human malignant peripheral nerve sheath tumors. AB - Patients with neurofibromatosis type 1 develop aggressive Schwann cell neoplasms known as malignant peripheral nerve sheath tumors (MPNSTs). Although tumor suppressor gene mutations play an important role in MPNST pathogenesis, it is likely that dysregulated signaling by as yet unidentified growth factors also contributes to the formation of these sarcomas. To test the hypothesis that neuregulin-1 (NRG-1) growth factors promote mitogenesis in MPNSTs, we examined the expression and action of NRG-1 in human MPNSTs and neurofibromas, the benign precursor lesions from which MPNSTs arise. Multiple alpha and beta transmembrane precursors from the class II and III NRG-1 subfamilies are present in both tumor types. Neoplastic Schwann cells within these neoplasms variably express the erbB kinases mediating NRG-1 responses (erbB2, erbB3 and/or erbB4). Human MPNST cell lines (Mash-1, YST-1, NMS-2 and NMS-2PC cells) similarly coexpress multiple NRG-1 isoforms and erbB receptors. These MPNST lines are NRG-1 responsive and demonstrate constitutive erbB phosphorylation. Treatment with PD168393 and PD158780, two structurally and mechanistically distinct erbB inhibitors, abolishes erbB phosphorylation and reduces DNA synthesis in these lines. These findings suggest that autocrine and/or paracrine NRG-1/erbB signaling promotes neoplastic Schwann cell proliferation and may be an important therapeutic target in neurofibromas and MPNSTs. PMID- 15897878 TI - Integrin alpha6beta4 promotes expression of autotaxin/ENPP2 autocrine motility factor in breast carcinoma cells. AB - In advanced breast carcinomas, the alpha6beta4 integrin is associated with a migratory and invasive phenotype. In our current study, we show that expression of the alpha6beta4 integrin in MDA-MB-435 breast carcinoma cells leads to increased expression of the autocrine motility factor autotaxin, as determined by Affymetrix gene chip, real-time quantitative RT-PCR and immunoblot analyses. We further demonstrate that increased autotaxin secretion from integrin alpha6beta4 expressing cells acts to enhance chemotaxis through its ability to convert lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA) and accounts for 80% of the motogenic activity of the conditioned medium. We determine that integrin alpha6beta4-dependent overexpression of autotaxin in MDA-MB-435 cells is mediated by NFAT1, but not NFAT5, through the use of siRNAs that specifically target autotaxin, integrin beta4, NFAT1 and NFAT5. Finally, we show by electrophoretic mobility shift assays that two consensus NFAT binding sites found in the autotaxin promoter strongly and specifically bind NFAT1 from integrin alpha6beta4 expressing cells. In summary, we find that the alpha6beta4 integrin potentiates autotaxin expression through the upregulation and activation of NFAT1. These observations highlight for the first time a mechanism by which NFAT transcription factors can facilitate an invasive and motile phenotype downstream of integrin alpha6beta4 signaling. PMID- 15897879 TI - G1/S transcriptional networks modulated by the HOX11/TLX1 oncogene of T-cell acute lymphoblastic leukemia. AB - The HOX11/TLX1 homeobox gene is aberrantly expressed in a subset of T-cell acute lymphoblastic leukemia (T-ALL). Here, we employed oligonucleotide microarrays to compare the expression profiles of the K3P and Sil leukemic cell lines originating from patients with HOX11+ T-ALL to that of Jurkat cells, which originated from a distinct subtype of T-ALL (TAL1+). To distinguish potential HOX11 target genes from those characteristic of the stage of HOX11 leukemic arrest, we also performed gene expression analysis on Jurkat cells, genetically engineered to express exogenous HOX11. The resulting HOX11 gene expression signature, which was validated for representative signaling pathways by transient transfection of reporter constructs, was characterized by elevated expression of transcriptional programs involved in cell proliferation, including those regulated by E2F, c-Myc and cAMP response element-binding protein. We subsequently showed that ectopic HOX11 expression resulted in hyperphosphorylation of the retinoblastoma protein (Rb), which correlated with inhibition of the major Rb serine/threonine phosphatase PP1. HOX11 also inhibited PP2A serine/threonine phosphatase activity concomitant with stimulation of the AKT/PKB signaling cascade. These results suggest that transcriptional deregulation of G1/S growth-control genes, mediated in large part through blockade of PP1/PP2A phosphatase activity, plays an important role in HOX11 pathobiology. PMID- 15897880 TI - All-trans retinoic acid treatment of Wilms tumor cells reverses expression of genes associated with high risk and relapse in vivo. AB - Wilms tumor is one of the most frequent neoplasias in children. Our previous microarray screening in a large series of Wilms tumors revealed several candidate genes that are deregulated in advanced tumors and are part of the retinoic acid signaling pathway. To investigate whether retinoic acid could be employed as a novel therapeutic agent in these tumors, we treated cultured Wilms tumor cells with different concentrations of all-trans retinoic acid (ATRA) and assessed gene expression changes by real-time RT-PCR as well as microarray analysis. Several genes like RARRES1, RARRES3, CTGF, CKS2, CCNA2, IGFBP3, UBE2C, CCL2 or ITM2B that were previously found to be deregulated in advanced tumors exhibited opposite expression changes after ATRA treatment. In addition to enhanced retinoid signaling, the transforming growth factor-beta (TGFbeta) pathway was strongly activated by ATRA treatment of Wilms tumor cells. Both the retinoic acid and the TGFbeta pathway mediate inhibition of cell growth. These findings represent the first molecular evidence of a potential benefit from ATRA treatment in Wilms tumors. PMID- 15897881 TI - DNA damage induce gamma-tubulin-RAD51 nuclear complexes in mammalian cells. AB - Rad51 protein plays an essential role in recombination repair of DNA double strand breaks and DNA crosslinking adducts. It is part of complexes which can vary with the stage of the cell cycle and the nature of the DNA lesions. During a search for Rad51-associated proteins in CHO nuclear extracts of S-phase cells by mass spectrometry of proteins immunoprecipitated with Rad51 antibodies, we identified a centrosomal protein, gamma-tubulin. This association was confirmed by the reverse immunoprecipitation with gamma-tubulin antibodies. Both proteins copurified from HeLa cells nuclear extracts following a tandem affinity purification of double-tagged Rad51. Immunofluorescence analysis showed colocalization of both Rad51 and gamma-tubulin in discrete foci in mammalian cell nuclei. The number of colocalized foci and their overlapping area increased in the presence of DNA damage produced by genotoxic treatments either during S phase or in exponentially growing cells. These variations did not result from an overall stress because microtubule cytoskeleton poisons devoid of direct interactions with DNA, such as taxol or colcemid, did not lead to an increase of this association. The recruitment of Rad51 and gamma-tubulin in the same nuclear complex suggests a link between DNA recombination repair and the centrosome function during the cell cycle. PMID- 15897882 TI - HIPK2 contributes to PCAF-mediated p53 acetylation and selective transactivation of p21Waf1 after nonapoptotic DNA damage. AB - The p53 tumor suppressor gene is activated in response to DNA damage resulting in either growth arrest or apoptosis. We previously demonstrated the specific involvement of homeodomain interacting protein-kinase 2 (HIPK2), a nuclear serine/threonine kinase, in inducing p53-dependent apoptosis through selective p53 phosphorylation at serine 46 after severe genotoxic damage. Here we show that HIPK2 contributes to p53 regulation, independently from serine 46 phosphorylation upon nonapoptotic DNA damage such as that induced by cytostatic doses of cisplatin. We show that HIPK2 depletion by RNA interference inhibits p53 binding to the p21Waf1 promoter affecting its p53-induced transactivation thereby allowing cell proliferation. We found that nonapoptotic DNA damage induces p53 acetylation mediated by the HAT protein PCAF and this p53 post-translational modification is abolished by HIPK2 depletion. In this regard, we found that HIPK2 cooperates with PCAF to induce selectively p53 transcriptional activity toward the p21Waf1 promoter while depletion of either HIPK2 or PCAF abolished this function. These data show that HIPK2 regulates the p53 growth arrest function through its PCAF-mediated acetylation. PMID- 15897883 TI - Gene expression profiling of cancer progression reveals intrinsic regulation of transforming growth factor-beta signaling in ErbB2/Neu-induced tumors from transgenic mice. AB - Upregulation of HER2/ErbB2/Neu occurs in 15-30% of human breast cancers and correlates with poor prognosis. Identification of ErbB2/Neu transcriptional targets should facilitate development of novel therapeutic approaches. Development of breast cancer is a multistep process; thus, to identify the transcriptomes associated with different stages of progression of tumorigenesis, we compared expression profiles of mammary tumors and preneoplastic mammary tissue from MMTV-Neu transgenic mice to expression profiles of wild-type mammary glands using Affymetrix microarrays. We identified 324 candidate genes that were unique to ErbB2/Neu-induced tumors relative to normal mammary gland tissue from wild-type controls. Expression of a subset of these genes (82) was also changed in the preneoplastic mammary glands compared to wild-type controls, indicating that they may play a pivotal role during early events of ErbB2/Neu-initiated mammary tumorigenesis. Further analysis of the microarray data revealed that expression of several known transforming growth factor (TGF)-beta target genes was altered, suggesting that the TGF-beta signaling cascade is downregulated in ErbB2/Neu-induced tumors. Western blot analysis for TGF-beta-Receptor-I/ALK5 and immunohistochemistry for TGF-beta-Receptor-I/ALK5 and phosphorylated/activated Smad2 confirmed that the Smad-dependent TGF-beta signaling cascade was inactive in these tumors. Although absent in most of the tumor, phosphorylated Smad2 was present in the periphery of tumors. Interestingly, presence of phosphorylated/activated Smad2 correlated with expression of Activin-Receptor IB/ALK4, suggesting that although Smad-dependent TGF-beta signaling is absent in ErbB2/Neu-induced tumors, Activin signaling may be active at the leading edge of these tumors. Cumulatively, these data indicate that the TGF-beta pathway is intrinsically suppressed in ErbB2/Neu tumors via a mechanism involving loss of TGF-beta-Receptor-I/ALK5. PMID- 15897884 TI - The MYO1F, unconventional myosin type 1F, gene is fused to MLL in infant acute monocytic leukemia with a complex translocation involving chromosomes 7, 11, 19 and 22. AB - We analysed a complex translocation involving chromosomes 7, 11, 19 and 22 in infant acute monocytic leukemia, and identified that the MLL gene on 11q23 was fused to the unconventional myosin type 1F, MYO1F, gene on 19p13.2-13.3. MYO1F consists of at least 28 exons and was predicted to encode a 1098-amino-acid with an N-terminal head domain containing both ATP-binding and actin-binding sequences, a neck domain with a single IQ motif, and a tail with TH1, TH2 and SH3 domains. Northern blot analysis of RNAs prepared from multiple tissues showed that the expression of approximately 4-kb transcripts appeared constant in most tissues examined. However, MYO1F was expressed in only three of 22 leukemic cell lines. The MLL-MYO1F fusion protein contains almost the entire MYO1F, however, C terminal MYO1F has neither the transactivation domain nor the dimerization domain found in various MLL fusion partners. Further analysis of this novel type of MLL fusion protein would provide new insights into leukemogenesis. MYO1F is the fourth partner gene of MLL on 19p13. At the cytogenetic level, it may be difficult to distinguish MLL-ENL, MLL-ELL, MLL-EEN and MLL-MYO1F fusions created by t(11;19)(q23;p13), and it is likely that cases of t(11;19) lacking a known fusion gene may result in this gene fusion. PMID- 15897885 TI - FGFR3 and Ras gene mutations are mutually exclusive genetic events in urothelial cell carcinoma. AB - Fibroblast growth factor receptor 3 (FGFR3) mutations are frequent in superficial urothelial cell carcinoma (UCC). Ras gene mutations are also found in UCC. As oncogenic activation of both FGFR3 and Ras is predicted to result in stimulation of the mitogen-activated protein kinase (MAPK) pathway, we hypothesized that these might be mutually exclusive events. HRAS mutation has been widely studied in UCC, but all three Ras gene family members have not been screened for mutation in the same sample series. We screened 98 bladder tumours and 31 bladder cell lines for mutations in FGFR3, HRAS, NRAS and KRAS2. FGFR3 mutations were present in 54 tumours (55%) and three cell lines (10%), and Ras gene mutations in 13 tumours (13%) and four cell lines (13%). These included mutations in all three Ras genes; ten in HRAS, four in KRAS2 and four in NRAS and these were not associated with either tumour grade or stage. In no cases were Ras and FGFR3 mutation found together. This mutual exclusion suggests that FGFR3 and Ras gene mutation may represent alternative means to confer the same phenotype on UCC cells. If these events have biological equivalence, Ras mutant invasive UCC may represent a novel subgroup. PMID- 15897886 TI - E2F-8: an E2F family member with a similar organization of DNA-binding domains to E2F-7. AB - E2F is a family of transcription factors implicated in cell cycle control. To understand the role of E2F in controlling cell cycle progression, it is necessary to clarify the breadth of the E2F family. To date, seven E2F subunits have been identified. We report here the characterization of a new E2F subunit, E2F-8, which resembles the organization of E2F-7 in the presence of two separate DNA binding domains, the integrity of which is required for E2F-8 to bind to DNA. Furthermore, like E2F-7, we find that E2F-8 can repress transcription and delay cell cycle progression. The similarities between E2F-7 and E2F-8 define a new subgroup of the E2F family, and further imply that E2F-7 and E2F-8 may act through overlapping mechanisms in mediating cell cycle control. PMID- 15897887 TI - Meta-analysis of microarray data on pancreatic cancer defines a set of commonly dysregulated genes. AB - Pancreatic ductal adenocarcinoma is the eighth most common cancer with the lowest overall 5-year relative survival rate of any tumor type today. Expression profiling using microarrays has been widely used to identify genes associated with pancreatic cancer development. To extract maximum value from the available gene expression data, we applied a meta-analysis to search for commonly differentially expressed genes in pancreatic ductal adenocarcinoma. We obtained data sets from four different gene expression studies on pancreatic cancer. We selected a consensus set of 2984 genes measured in all four studies and applied a meta-analysis approach to evaluate the combined data. Of the genes identified as differentially expressed, several were validated using RT-PCR and immunohistochemistry. Additionally, we used a class discovery algorithm to identify a gene expression signature. Our meta-analysis revealed that the pancreatic cancer gene expression data sets shared a significant number of up- and downregulated genes, independent of the technology used. This interstudy crossvalidation approach generated a set of 568 genes that were consistently and significantly dysregulated in pancreatic cancer. Of these, 364 (64.1%) were upregulated and 204 (35.9%) were downregulated in pancreatic cancer. Only 127 (22%) were described in the published individual analyses. Functional annotation of the genes revealed that genes presumably associated with the cell adhesion mediated drug resistance pathway are frequently overexpressed in pancreatic cancer. Meta-analysis is an important tool for the identification and validation of differentially expressed genes. These could represent good candidates for novel diagnostic and therapeutic approaches to pancreatic cancer. PMID- 15897888 TI - Upregulation of VEGF-C by androgen depletion: the involvement of IGF-IR-FOXO pathway. AB - Androgen ablation therapy is eventually followed by a more metastatic and androgen-refractory stage of prostate cancer. The detailed molecular mechanism of this gradual transition is not clearly understood. Recent reports correlate the high abundance of vascular endothelial growth factor-C (VEGF-C) to the lymph node metastasis seen in human prostate cancer (Tsurusaki et al., 1999). In this study, we report that androgen ablation in LNCaP cells augment the transcriptional upregulation of VEGF-C and the downregulation of the IGF-IR pathway, due to androgen withdrawal, is a potential mechanism for this observed VEGF-C transcription. Forkhead transcription factor FOXO-1, activated by SIRT-1, was identified as the downstream molecule within this pathway. Furthermore, the VEGF C-induced increase of Bag-IL expression in LNCaP cells suggests that VEGF-C plays a role in the androgen-independent reactivation of the androgen receptor, resulting in androgen-refractory prostate cancer growth. PMID- 15897889 TI - Gene expression responses to DNA damage are altered in human aging and in Werner Syndrome. AB - The accumulation of DNA damage and mutations is considered a major cause of cancer and aging. While it is known that DNA damage can affect changes in gene expression, transcriptional regulation after DNA damage is poorly understood. We characterized the expression of 6912 genes in human primary fibroblasts after exposure to three different kinds of cellular stress that introduces DNA damage: 4-nitroquinoline-1-oxide (4NQO), gamma-irradiation, or UV-irradiation. Each type of stress elicited damage specific gene expression changes of up to 10-fold. A total of 85 genes had similar changes in expression of 3-40-fold after all three kinds of stress. We examined transcription in cells from young and old individuals and from patients with Werner syndrome (WS), a segmental progeroid condition with a high incidence of cancer, and found various age-associated transcriptional changes depending upon the type of cellular stress. Compared to young individuals, both WS and old individuals had similarly aberrant transcriptional responses to gamma- and UV-irradiation, suggesting a role for Werner protein in stress-induced gene expression. Our results suggest that aberrant DNA damage-induced gene regulation may contribute to the aging process and the premature aging in WS. PMID- 15897890 TI - Silencing of the Tropomyosin-1 gene by DNA methylation alters tumor suppressor function of TGF-beta. AB - Loss of actin stress fibers has been associated with cell transformation and metastasis. TGF-beta induction of stress fibers in epithelial cells requires high molecular weight tropomyosins encoded by TPM1 and TPM2 genes. Here, we investigated the mechanism underlying the failure of TGF-beta to induce stress fibers and inhibit cell migration in metastatic cells. RT-PCR analysis in carcinoma cell lines revealed a significant reduction in TPM1 transcripts in metastatic MDA-MB-231, MDA-MB-435 and SW620 cell lines. Treatment of these cells with demethylating agent 5-aza-2'-deoxycytidine (5-aza-dC) increased mRNA levels of TPM1 with no effect on TPM2. Importantly, 5-aza-dC treatment of MDA-MB-231 cells restored TGF-beta induction of TPM1 and formation of stress fibers. Forced expression of TPM1 by using Tet-Off system increased stress fibers in MDA-MB-231 cells and reduced cell migration. A potential CpG island spanning the TPM1 proximal promoter, exon 1, and the beginning of intron 1 was identified. Bisulfite sequencing showed significant cytosine methylation in metastatic cell lines that correlated with a reduced expression of TPM1. Together these results suggest that epigenetic suppression of TPM1 may alter TGF-beta tumor suppressor function and contribute to metastatic properties of tumor cells. PMID- 15897891 TI - Role of type I receptors for anti-Mullerian hormone in the SMAT-1 Sertoli cell line. AB - Anti-Mullerian hormone (AMH) is a member of the transforming growth factor-beta family responsible for regression of Mullerian ducts during male sexual differentiation and for regulation of gonadal steroidogenesis. AMH is also a gonadal tumor suppressor which mediates its effects through a specific type II receptor and the bone morphogenetic protein (BMP)-specific Smad proteins, suggesting that AMH and BMPs could also share type I receptors, namely activin like kinases (ALKs)2, 3 or 6. However, attempts to identify a unique AMH type I receptor among them were unsuccessful. Here, using kinase-deficient type I receptors and small interfering RNA technology, we demonstrate that, in an AMH Sertoli target cell line, ALK3 mediates AMH effects on both Smad1 activation and P450 side-chain cleavage enzyme. In addition, transfecting a combination of normal and kinase-deficient receptors, we show that ALK2 can compensate for the absence of ALK3 and probably acts in synergy with ALK3 at high concentrations of AMH to activate Smad1, whereas ALK6 has a competitive inhibitory effect. These results are a first step in understanding how AMH transduces its effects in immature Sertoli cells. PMID- 15897892 TI - The MCT-1 oncogene product impairs cell cycle checkpoint control and transforms human mammary epithelial cells. AB - Multiple copies in T-cell maligancy (MCT-1) is a putative oncogene initially identified in a human T-cell lymphoma. Forced expression of MCT-1 has recently been shown to induce cell transformation and proliferation, as well as to activate survival-related PI-3K/AKT pathways protecting cells from apoptosis. MCT 1 protein is stabilized in response to DNA damage. The impact of MCT-1 overexpression on DNA damage response remains unknown. Here, we show that MCT-1 deregulates cell cycle checkpoints. The phosphorylation of genomic stabilizers H2AX and NBS1 are enhanced in MCT-1-overexpressing cells. Forced expression of MCT-1 significantly increases the number of DNA damage-induced foci involving gamma-H2AX and 53BP1. In MCT-1-overexpressing cells, the proportion of S-phase cell population is preferentially increased after exposure to gamma-irradiation compared to controls. Knockdown of endogenous MCT-1 using an siRNA approach attenuates the H2AX phosphorylation and the G1/S checkpoint defect. Furthermore, MCT-1 is capable of transforming immortalized human mammary epithelial cells and promoting genomic instability. These data shed light on the role of MCT-1 in the cellular response to DNA damage and its involvement in malignant transformation. PMID- 15897893 TI - TCDD induces c-jun expression via a novel Ah (dioxin) receptor-mediated p38-MAPK dependent pathway. AB - The aryl hydrocarbon receptor (AhR) has a fundamental role during postnatal liver development and is essential for mediating dioxin toxicity. However, the genetic programs mediating, both, the toxic and physiological effects downstream of the transcription factor AhR are in major parts unknown. We have identified the proto oncogene c-jun as a novel target gene of AhR. Induction of c-jun depends on activation of p38-mitogen-activated protein kinase (MAPK) by an AhR-dependent mechanism. None of the kinases that are known to phosphorylate p38-MAPK is activated by AhR. Neither the dephosphorylation rate of p38-MAPK is reduced. Furthermore, increased p38-MAPK phosphorylation in response to dioxins does not require ongoing transcription. These findings establish activating 'cross-talk' with MAPK signaling as a novel principle of AhR action, which is apparently independent of the AhR's function as a DNA-binding transcriptional activator. PMID- 15897894 TI - Molecular cloning and characterization of STAMP2, an androgen-regulated six transmembrane protein that is overexpressed in prostate cancer. AB - We have identified a novel gene, six transmembrane protein of prostate 2 (STAMP2), named for its high sequence similarity to the recently identified STAMP1 gene. STAMP2 displays a tissue-restricted expression with highest expression levels in placenta, lung, heart, and prostate and is predicted to code for a 459-amino acid six transmembrane protein. Using a form of STAMP2 labeled with green flourescent protein (GFP) in quantitative time-lapse and immunofluorescence confocal microscopy, we show that STAMP2 is primarily localized to the Golgi complex, trans-Golgi network, and the plasma membrane. STAMP2 also localizes to vesicular-tubular structures in the cytosol and colocalizes with the Early Endosome Antigen1 (EEA1) suggesting that it may be involved in the secretory/endocytic pathways. STAMP2 expression is exquisitely androgen regulated in the androgen-sensitive, androgen receptor-positive prostate cancer cell line LNCaP, but not in androgen receptor-negative prostate cancer cell lines PC-3 and DU145. Analysis of STAMP2 expression in matched normal and tumor samples microdissected from prostate cancer specimens indicates that STAMP2 is overexpressed in prostate cancer cells compared with normal prostate epithelial cells. Furthermore, ectopic expression of STAMP2 in prostate cancer cells significantly increases cell growth and colony formation suggesting that STAMP2 may have a role in cell proliferation. Taken together, these data suggest that STAMP2 may contribute to the normal biology of the prostate cell, as well as prostate cancer progression. PMID- 15897895 TI - Interaction and colocalization of Rad9/Rad1/Hus1 checkpoint complex with replication protein A in human cells. AB - Replication protein A (RPA) is a eukaryotic single-stranded DNA-binding protein consisting of three subunits of 70-, 32-, and 14-kDa (RPA70, RPA32, RPA14, respectively). It is a protein essential for most cellular DNA metabolic pathways. Checkpoint proteins Rad9, Rad1, and Hus1 form a clamp-like complex which plays a central role in the DNA damage-induced checkpoint response. In this report, we presented the evidence that Rad9-Rad1-Hus1 (9-1-1) complex directly interacted with RPA in human cells, and this interaction was mediated by the binding of Rad9 protein to both RPA70 and RPA32 subunits. In addition, the cellular interaction of 9-1-1 with RPA or hyperphosphorylated RPA was stimulated by UV irradiation or camptothecin treatment in a dose-dependent manner. Such treatments also resulted in the colocalization of the nuclear foci formed with the two complexes. Consistently, knockdown of the RPA expression in cells by the small interference RNA (siRNA) blocked the DNA damage-dependent chromatin association of 9-1-1, and also inhibited the 9-1-1 complex formation. Taken together, our results suggest that 9-1-1 and RPA complexes collaboratively function in DNA damage responses, and that the RPA may serve as a regulator for the activity of 9-1-1 complex in the cellular checkpoint network. PMID- 15897896 TI - Hepatitis C virus core triggers apoptosis in liver cells by inducing ER stress and ER calcium depletion. AB - Hepatitis C virus (HCV) core, known to be involved in liver carcinogenesis, is processed in the endoplasmic reticulum (ER). We thus investigated the impact of three HCV core isolates on ER stress, ER calcium signalling and apoptosis. We show that HCV core constructs trigger hyperexpression of Grp78/BiP, Grp 94, calreticulin and sarco/endoplasmic reticulum calcium ATPase, inducing ER stress. By using the ER-targeted aequorin calcium probe, we found that ER calcium depletion follows ER stress in core-expressing cells. HCV core induces apoptosis through overexpression of the CHOP/GADD153 proapoptotic factor, Bax translocation to mitochondria, mitochondrial membrane depolarization, cytochrome c release, caspase-3 and PARP cleavage. Furthermore, reversion of HCV core-induced ER calcium depletion (by transfection of SERCA2) completely abolished mitochondrial membrane depolarization, suggesting that both ER stress (through CHOP overexpression) and calcium signalling play a major role in the HCV core-mediated control of apoptosis. ER stress and apoptosis were also found in a proportion of HCV-full-length replicon-expressing cells and in the liver of HCV core transgenic mice. In conclusion, our data demonstrate that HCV core deregulates the control of apoptosis by inducing ER stress and ER calcium depletion providing new elements to understand the mechanisms involved in HCV-related liver chronic diseases. PMID- 15897897 TI - Identification of midkine as a mediator for intercellular transfer of drug resistance. AB - Resistance to cytotoxic agents is a major limitation for their clinical use to treat human cancers. Tumors become resistant to chemotherapy when a subset of cells undergoes molecular changes leading to overexpression of drug transport proteins, alterations in drug-target interactions or reduced ability to commit apoptosis. However, such changes may not be sufficient to explain why both resistant and nonresistant cells survive drug's action in tumors that ultimately become drug resistant. We hypothesized that, in such tumors, a cytoprotective relationship may exist between drug-resistant and neighboring drug-sensitive cells. The present study addresses the possibility that drug-resistant cells secrete in their culture medium factors able to protect sensitive cells from drug toxicity. A survival molecule, midkine, was identified by cDNA array to be expressed only in drug-resistant cells. Midkine-enriched fractions obtained by affinity chromatography exert a significant cytoprotective effect against doxorubicin in the wild-type drug-sensitive cells. Moreover, transfection of these cells with the midkine gene caused a decreased response to doxorubicin. The underlying mechanism of this cytoprotection appeared to imply activation of the Akt pathway and inhibition of drug-induced proliferation arrest as well as apoptotic cell death. These findings provide evidence for the existence of intercellular cytoprotective signals such as the one mediated by midkine, originating from cells with acquired drug resistance to protect neighboring drug sensitive cells and thus contribute to development of resistance to chemotherapy. PMID- 15897898 TI - The Human mineral dust-induced gene, mdig, is a cell growth regulating gene associated with lung cancer. AB - Environmental or occupational exposure to mineral dusts, mainly silica and asbestos, is associated with an increased incidence of lung inflammation, fibrosis, and/or cancer. To better understand the molecular events associated with these pulmonary diseases, we attempted to identify genes that are regulated by mineral dusts. Using a differential display reverse transcription polymerase chain reaction technique and mRNAs of alveolar macrophages from both normal individuals and coal miners, we identified a novel mineral dust-induced gene named mdig, which had not been fully characterized. The expression of mdig mRNA was detected in alveolar macrophages from coal miners but not from normal subjects. The inducible expression of mdig could be observed in A549 cells exposed to silica particles in a time-dependent manner. The full-length mdig mRNA was expressed in human lung cancer tissues but was barely detectable in the adjacent normal tissues. In addition, a number of lung cancer cell lines constitutively express mdig. Alternative spliced transcripts of mdig were detected in some lung cancer cell lines. Silencing mdig mRNA expression in A549 lung cancer cells by siRNA-mediated RNA interference inhibits cell proliferation and sensitizes the cells to silica-induced cytotoxicity. These results suggest that the mdig gene may be involved in the regulation of cell growth and possibly the development of cancer. PMID- 15897899 TI - Estrogen-induced G1/S transition of G0-arrested estrogen-dependent breast cancer cells is regulated by mitochondrial oxidant signaling. AB - We previously reported that 17-beta-estradiol (E2)-induced mitochondrial reactive oxygen species (mtROS) act as signaling molecules. The purpose of this study was to investigate the effects of E2-induced mtROS on cell cycle progression. E2 induced cell growth was reduced by antioxidants N-acetyl-L-cysteine (NAC), catalase, and the glutathione peroxidase mimic ebselen. Flow cytometry showed that mitochondrial blockers of protein synthesis (chloramphenicol), transcription and replication (ethidium bromide), and function (rotenone, rhodamine 6G) blocked E2-induced G1 to S transition. Reduction of E2-induced DNA synthesis in the presence of mitochondrial blockers occurred without influencing the level of ATP. Additionally, the mitochondrial blockers inhibited the E2-induced expression of early cell cycle genes such as cyclins D1, D3, E1, E2, and B2. NAC or rotenone reduced E2-induced cyclin D1 expression. Furthermore, E2-induced binding of AP-1 and CREB to the TRE and CRE response sequences, respectively, in the promoter of cyclin D1 was inhibited by NAC or rotenone. In addition, E2-induced expression of PCNA, PRC1, and bcl-2 were inhibited by mitochondrial blockers. These data indicate that E2-induced mtROS are involved in the regulation of early G1-phase progression. Since neither antioxidants nor mitochondrial blockers used in this study are reported to bind the estrogen receptor (ER), our findings suggest that E2-induced mtROS modulates G1 to S transition and some of the early G1 genes through a nongenomic, ER-independent signaling pathway. Thus, our results suggest (1) a new paradigm that estrogen-induced mitochondrial oxidants control the early stage of cell cycle progression and (2) provide the basis for the discovery of novel antioxidant-based drugs or antioxidant gene therapies for the prevention and treatment of estrogen-dependent breast cancer. PMID- 15897900 TI - Pituitary tumour transforming gene (PTTG) induces genetic instability in thyroid cells. AB - Cancer reflects the progressive accumulation of genetic alterations and subsequent genetic instability of cells. Cytogenetic studies have demonstrated the importance of aneuploidy in differentiated thyroid cancer development. The pituitary tumour transforming gene (PTTG), also known as securin, is a mitotic checkpoint protein which inhibits sister chromatid separation during mitosis. PTTG is highly expressed in many cancers and overexpression of PTTG induces aneuploidy in vitro. Using fluorescent intersimple sequence repeat PCR (FISSR PCR), we investigated the relationship between PTTG expression and the degree of genetic instability in normal and tumorous thyroid samples. The genomic instability index (GI index) was 6.7-72.7% higher in cancers than normal thyroid tissues. Follicular thyroid tumours exhibited greater genetic instability than papillary tumours (27.6% (n=9) versus 14.5% (n=10), P=0.03). We also demonstrated a strong relationship between PTTG expression and the degree of genetic instability in thyroid cancers (R2=0.80, P=0.007). To further investigate PTTG's role in genetic instability, we transfected FTC133 thyroid follicular cells and observed increased genetic instability in cells overexpressing PTTG compared with vector-only-transfected controls (n=3, GI Index VO=29.7+/-5.2 versus PTTG=63.7+/ 6.4, P=0.013). Further, we observed a dose response in genetic instability and PTTG expression (GI Index low dose (0.5 microg DNA/ six-well plate) PTTG=15.3%+/ 1.7 versus high dose (3 microg DNA) PTTG=50.8%+/-3.3, P=0.006). Overall, we describe the first use of FISSR-PCR in human cancers, and demonstrate that PTTG expression correlates with genetic instability in vivo, and induces genetic instability in vitro. We conclude that PTTG may be an important gene in the mutator phenotype development in thyroid cancer. PMID- 15897901 TI - Regulation of the phosphorylation and integrity of protein synthesis initiation factor eIF4GI and the translational repressor 4E-BP1 by p53. AB - Activation of a temperature-sensitive form of mouse p53 in murine erythroleukaemia cells rapidly inhibits protein synthesis and causes early dephosphorylation and cleavage of protein synthesis initiation factor eIF4GI and the eIF4E-binding protein 4E-BP1. Dephosphorylated 4E-BP1 and the cleaved products of 4E-BP1 and eIF4GI associate with eIF4E under these conditions, concomitant with decreased interaction of full-length eIF4GI with eIF4E. These changes may play an important role in preventing formation of the eIF4F complex and thus the initiation of protein synthesis. As observed previously for eIF4GI, the cleavage of 4E-BP1 is insensitive to the general caspase inhibitor z-VAD.FMK, consistent with a caspase-independent mechanism of factor modification and regulation of protein synthesis. Comparison of the p53-induced patterns of eIF4GI and 4E-BP1 dephosphorylation and cleavage with those caused by the mTOR inhibitor rapamycin indicates that p53 activation and rapamycin have distinct but additive effects. Moreover, p53 activation inhibits rapamycin-insensitive protein kinase activity against 4E-BP1. P53 and rapamycin have additive effects on the inhibition of overall protein synthesis. These data suggest that the inhibition of protein synthesis by p53 is largely independent of the regulation of rapamycin sensitive mTOR in the system under investigation. PMID- 15897902 TI - KIF14 is a candidate oncogene in the 1q minimal region of genomic gain in multiple cancers. AB - Gain of chromosome 1q31-1q32 is seen in >50% of retinoblastoma and is common in other tumors. To define the minimal 1q region of gain, we determined genomic copy number by quantitative multiplex PCR of 14 sequence tagged sites (STSs) spanning 1q25.3-1q41. The most frequently gained STS at 1q32.1 (71%; 39 of 55 retinoblastoma) defined a 3.06 Mbp minimal region of gain between flanking markers, containing 14 genes. Of these, only KIF14, a putative chromokinesin, was overexpressed in various cancers by real-time RT-PCR. KIF14 mRNA was expressed in 20/22 retinoblastoma samples 100-1000-fold higher than in retina (t-test P=0.00002); cell lines (n=10) had higher levels than tumors (n=12) (P=0.009). KIF14 protein was overexpressed in retinoblastoma tumors and breast cancer cell lines by immunoblot. KIF14 was expressed in 4/4 breast cancer cell lines 31-92 fold higher than in normal breast tissue, in 5/5 medulloblastoma cell lines 22-79 fold higher than in fetal brain, and in 10/22 primary lung tumors 3-34-fold higher than in normal lung. Patients with lung tumors that overexpress KIF14 showed a trend toward decreased survival. KIF14 may thus be important in oncogenesis, and has promise as a prognostic indicator and therapeutic target. PMID- 15897903 TI - Accumulation of hypoxia-inducible factor-1alpha is limited by transcription dependent depletion. AB - In the presence of oxygen and iron, hypoxia-inducible factor (HIF-1alpha) is rapidly degraded via the prolyl hydroxylases (PHD)/VHL pathways. Given striking similarities between p53 and HIF-1alpha regulation, we previously suggested that HIF-1 transcriptionally initiates its own degradation and therefore inhibitors of transcription must induce HIF-1alpha. Under normoxia, while inducing p53, inhibitors of transcription did not induce HIF-1alpha. Under hypoxia or low iron (DFX), inhibitors of transcription dramatically super-induced HIF-1alpha. Removal of inhibitors resulted in outburst of the HIF-1-dependent transcription followed by depletion of HIF-1alpha. Although hypoxia/DFX induced PHD3, we excluded the PHD/VHL pathway in the regulation of HIF-1alpha under hypoxia/DFX. The transcription-dependent degradation of HIF-1alpha under hypoxia occurs via the proteasome and is accelerated by protein acetylation. Thus, HIF-1alpha is regulated by two distinct mechanisms. Under normoxia, HIF-1alpha is degraded via the classic PHD/VHL pathway, is expressed at low levels and therefore does not activate the feedback loop. But under hypoxia, HIF-1alpha accumulates and transcriptionally activates its own degradation that is independent from the PHD/VHL pathway. PMID- 15897904 TI - Paclitaxel induces the phosphorylation of the eukaryotic translation initiation factor 4E-binding protein 1 through a Cdk1-dependent mechanism. AB - Initial chemotherapeutic treatment triggers a stress-related response, which can lead to an increase in the expression of survival proteins. In this study we examine whether paclitaxel (PTX) alters the expression and/or phosphorylation of the translation initiation proteins, eukaryotic initiation factor 4E (eIF-4E) and 4E-binding protein (4E-BP1), a suppressor of eIF-4E in the dephosphorylated state. We found that PTX induced the hyperphosphorylation of 4E-BP1 in the breast cancer cell line, MDA MB 231, which reduced its association with eIF-4E, but did not alter the expression and phosphorylation of eIF-4E. The hyperphosphorylation of 4E-BP1 correlated with G2/M accumulation and with an increase in the phosphorylation of cdk1 substrates. Cotreatment with a histone deacetylase inhibitor (an indirect inhibitor of cdk activity), purvalanol A and roscovitine (direct cdk inhibitors), and the reduction of cyclin B expression using RNA interference decreased the hyperphosphorylation of 4E-BP1 in PTX treated cells. The hyperphosphorylation of 4E-BP1 by PTX increased the association of eIF-4E with eIF-4G, whereas cotreatment with purvalanol A inhibited the association of eIF-4E with eIF-4G in PTX treated cells. Taken together, our data suggest that PTX-increases the functional level of eIF-4E by promoting the hyperphosphorylation and release of 4E-BP1 through a cdk1-dependent mechanism. PMID- 15897905 TI - Multiple defects of the antigen-processing machinery components in human neuroblastoma: immunotherapeutic implications. AB - Low expression of human leukocyte antigen (HLA) class I in human tumors may be related to defects of the antigen-processing machinery (APM) components. Neuroblastoma cells are virtually HLA class I negative, but (i) the underlying mechanisms are unknown, and (ii) expression of the APM components has never been investigated. Here we have used a panel of novel monoclonal antibodies to proteasomal and immunoproteasomal components, chaperons and transporter associated with antigen processing (TAP) to characterize 24 stroma-poor neuroblastoma tumors and six neuroblastoma cell lines. Primary tumors showed defects in the expression of zeta, tapasin, TAP1 or TAP2, HLA class I heavy chain and beta2 microglobulin, LMP2 and LMP7, as compared to normal adrenal medulla. Neuroblastoma cell lines displayed roughly similar patterns of APM expression in comparison to primary tumors. Incubation of neuroblastoma cell lines with interferon-gamma caused upregulation of HLA class I molecules and reduced lysis by killer inhibitory receptor HLA ligand-matched NK cells. Defects in APM components explain reduced peptide loading on HLA class I molecules, their instability and failure to be expressed on the cell surface. HLA class I upregulation by interferon-gamma, although enhancing neuroblastoma cell recognition by cytotoxic T cells, dampens their susceptibility to NK cells. PMID- 15897906 TI - HDAC inhibitors enhance the apoptosis-inducing potential of TRAIL in breast carcinoma. AB - Histone deacetylase (HDAC) inhibitors induce differentiation and/or apoptosis in a variety of cell types by activating transcription of target genes. Activation of the death receptor (DR) pathway by tumor necrosis factor-related apoptosis inducing ligand (TRAIL) induces apoptosis preferentially in cancer cells. Here, we investigated the intracellular mechanisms by which HDAC inhibitors (suberoylanilide hydroxamic acid, m-carboxycinnamic acid bis-hydroxamide, MS-275 and trichostatin A) enhance the apoptosis-inducing potential of TRAIL in breast cancer cells in vitro. A synergism in apoptosis was observed in both TRAIL sensitive and -resistant cells upon sequential treatments with HDAC inhibitors followed by TRAIL. HDAC inhibitors synergized with TRAIL by inducing DRs DR4/TRAIL-R1 and DR5/TRAIL-R2 through NFkappaB activation and some of the proapoptotic members of the Bcl-2 family, and engaging the mitochondrial pathway. The ability of HDAC inhibitors to sensitize TRAIL-resistant cells suggests that HDAC inhibitors may induce fundamental alterations in cell signaling pathways. Thus, the sequential treatments with HDAC inhibitors followed by TRAIL may be used as a new therapeutic approach for the treatment of human cancers. PMID- 15897907 TI - Identification of molecular apocrine breast tumours by microarray analysis. AB - Previous microarray studies on breast cancer identified multiple tumour classes, of which the most prominent, named luminal and basal, differ in expression of the oestrogen receptor alpha gene (ER). We report here the identification of a group of breast tumours with increased androgen signalling and a 'molecular apocrine' gene expression profile. Tumour samples from 49 patients with large operable or locally advanced breast cancers were tested on Affymetrix U133A gene expression microarrays. Principal components analysis and hierarchical clustering split the tumours into three groups: basal, luminal and a group we call molecular apocrine. All of the molecular apocrine tumours have strong apocrine features on histological examination (P=0.0002). The molecular apocrine group is androgen receptor (AR) positive and contains all of the ER-negative tumours outside the basal group. Kolmogorov-Smirnov testing indicates that oestrogen signalling is most active in the luminal group, and androgen signalling is most active in the molecular apocrine group. ERBB2 amplification is commoner in the molecular apocrine than the other groups. Genes that best split the three groups were identified by Wilcoxon test. Correlation of the average expression profile of these genes in our data with the expression profile of individual tumours in four published breast cancer studies suggest that molecular apocrine tumours represent 8-14% of tumours in these studies. Our data show that it is possible with microarray data to divide mammary tumour cells into three groups based on steroid receptor activity: luminal (ER+ AR+), basal (ER- AR-) and molecular apocrine (ER- AR+). PMID- 15897908 TI - Interaction of human Thy-1 (CD 90) with the integrin alphavbeta3 (CD51/CD61): an important mechanism mediating melanoma cell adhesion to activated endothelium. AB - The expression of the alphavbeta3 integrin (CD51/CD61) on human melanoma cells has been shown to be associated most closely with tumor progression and metastases formation in melanoma. Here, we demonstrated a specific interaction of the alphavbeta3 integrin on melanoma cells with the human Thy-1, an inducible cell adhesion molecule expressed on the cell surface of activated endothelial cells (EC). The interaction was shown by the binding of purified Thy-1 protein to alpha(V)beta(3) transfected cells, to alphavbeta3-expressing melanoma cells and to purified alpha(V)beta(3) integrin. Moreover, melanoma cells adhere specifically to Thy-1 transfectants via alphavbeta3 on melanoma cells showing the functional relevance of this interaction for cell adhesion. Finally, the importance of the alphavbeta3/Thy-1 interaction for the adhesion of melanoma cells to the activated endothelium was confirmed under static and flow conditions by the inhibition of melanoma cell adhesion to and transmigration across activated EC by blocking the alphavbeta3/Thy-1 interaction. In conclusion, we have identified a new pair of adhesion molecules Thy-1 and alphavbeta3 mediating the interaction of melanoma cells and activated EC. These data explain at least in part the high tumorigenicity of alphavbeta3-expressing melanoma cells and the association of alphavbeta3-positive melanoma cells with a high risk of metastasis and poor prognosis. PMID- 15897909 TI - FAS expression inversely correlates with PTEN level in prostate cancer and a PI 3 kinase inhibitor synergizes with FAS siRNA to induce apoptosis. AB - Fatty acid synthase (FAS), a key enzyme of the fatty acid biosynthetic pathway, has been shown to be overexpressed in various types of human cancer and is, therefore, considered to be an attractive target for anticancer therapy. However, the exact mechanism of overexpression of the FAS gene in tumor cells is not well understood. In this report, we demonstrate that the expression of the tumor suppressor gene PTEN has a significant inverse correlation with FAS expression in the case of prostate cancer in the clinical setting, and inhibition of the PTEN gene leads to the overexpression of FAS in vitro. We also found that the combination of the expression status of these two genes is a better prognostic marker than either gene alone. Furthermore, our results indicate that the specific inhibition of FAS gene by siRNA leads to apoptosis of prostate tumor cells, and inhibition of PI 3-kinase pathway synergizes with FAS siRNA to enhance tumor cell death. These results provide a strong rationale for exploring the therapeutic use of an inhibitor of the PTEN signaling pathway in conjunction with the FAS siRNA to inhibit prostate tumor growth. PMID- 15897910 TI - Aberrant hypermethylation of ID4 gene promoter region increases risk of lymph node metastasis in T1 breast cancer. AB - ID4 gene is a member of the inhibitor of DNA-binding (ID) family, which inhibits DNA binding of basic helix-loop-helix transcription factors. Certain human primary breast cancers reportedly have low or no expression of ID4 protein, but its role in carcinogenesis and cancer progression is unknown. To determine its possible role, we examined epigenetic inactivation of ID4 gene by promoter hypermethylation in human breast cell lines and T1 breast cancer tissues. Methylation status of ID4 promoter CpG island was assessed by methylation specific PCR (MSP); ID4 mRNA level was assessed by quantitative real-time RT-PCR. Of eight cell lines, two were fully methylated, four were partially methylated, and two were not methylated. ID4 mRNA level was suppressed in fully methylated cell lines. ID4 hypermethylation was observed in 16 of 24 (67%) node-positive and seven of 36 (19%) node-negative T1 primary breast cancers matched by patient age and tumor diameter. It was a significant risk factor for nodal metastasis (OR 13.1, P=0.0004). ID4 mRNA level was suppressed in hypermethylated cancer specimens (P=0.014). ID4 may play an important suppressive role in tumor progression, and its silencing by hypermethylation may increase the risk of regional lymph node metastasis. PMID- 15897911 TI - Cytotoxic T cell targeting of TRP-2 sensitizes human malignant glioma to chemotherapy. AB - Tyrosinase-related protein (TRP)-2 is not only expressed on glioma cells, but is naturally processed and presented by their surface MHC molecules and is recognized by TRP-2-specific cytotoxic T cells. After active immunotherapy, we detected TRP-2-specific cytotoxic T lymphocyte (CTL) activity in patients' peripheral blood mononuclear cells (PBMC). Tumor cells from postvaccination resections showed significantly lower TRP-2 expression and higher sensitivity to carboplatin and temozolomide than those autologous cell lines from prevaccination resections in two patients who demonstrated CTL response to TRP-2. One of two patients underwent treatment with temozolomide after recurrence and responded dramatically. TRP-2-transfected cell line (TRP-2-U373) resulted in significant drug resistance to carboplatin and temozolomide compared to wild-type U-373 (W U373). There was no significant difference, however, in the mRNA expression of other common drug resistance related proteins, such as BCRP-1, MGMT, MDR-1, MRP-1 and MRP-3, after TRP-2 transfection. TRP-2-U373 tumor cells were immunoselected by a TRP-2-specific CTL line. The immunoselected cells (IS-TRP-2-U373) demonstrated significantly increased sensitivity to carboplatin and temozolomide compared to TRP-2-U373. For the first time, we provide evidence that immunological targeting of tumor-associated antigen TRP-2 significantly increases sensitivity to chemotherapy. PMID- 15897912 TI - Human Cripto-1 overexpression in the mouse mammary gland results in the development of hyperplasia and adenocarcinoma. AB - Human Cripto-1 (CR-1) is overexpressed in approximately 80% of human breast, colon and lung carcinomas. Mouse Cr-1 upregulation is also observed in a number of transgenic (Tg) mouse mammary tumors. To determine whether CR-1 can alter mammary gland development and/or may contribute to tumorigenesis in vivo, we have generated Tg mouse lines that express human CR-1 under the transcriptional control of the mouse mammary tumor virus (MMTV). Stable Tg MMTV/CR-1 FVB/N lines expressing different levels of CR-1 were analysed. Virgin female MMTV/CR-1 Tg mice exhibited enhanced ductal branching, dilated ducts, intraductal hyperplasia, hyperplastic alveolar nodules and condensation of the mammary stroma. Virgin aged MMTV/CR-1 Tg mice also possessed persistent end buds. In aged multiparous MMTV/CR 1 mice, the hyperplastic phenotype was most pronounced with multifocal hyperplasias. In the highest CR-1-expressing subline, G4, 38% (12/31) of the multiparous animals aged 12-20 months developed hyperplasias and approximately 33% (11/31) developed papillary adenocarcinomas. The long latency period suggests that additional genetic alterations are required to facilitate mammary tumor formation in conjunction with CR-1. This is the first in vivo study that shows hyperplasia and tumor growth in CR-1-overexpressing animals. PMID- 15897913 TI - Inhibiting angiogenesis and tumorigenesis by a synthetic molecule that blocks binding of both VEGF and PDGF to their receptors. AB - Angiogenesis depends on vascular endothelial growth factor (VEGF) for initiation and platelet-derived growth factor (PDGF) for maintenance of blood vessels. We have designed a targeted library of compounds from which we identified a novel molecule, GFB-204, that binds PDGF and VEGF, blocks binding of PDGF and VEGF to their receptors (200-500 nM) and subsequently inhibits PDGFR and Flk-1 tyrosine phosphorylation and stimulation of the protein kinases Erk1, Erk2 and Akt and the signal transducer and activator of transcription STAT3. GFB-204 is selective for PDGF and VEGF and does not inhibit EGF, IGF-1 and FGF stimulation of Erk1/2, Akt and STAT3. GFB-204 inhibits endothelial cell migration and capillary network formation in vitro. Finally, treatment of mice with GFB-204 suppresses human tumor growth and angiogenesis. Thus, inhibition of VEGF and PDGF receptor binding with a synthetic molecule results in potent inhibition of angiogenesis and tumorigenesis. PMID- 15897914 TI - Metastatic prostate cancer presenting as an asymptomatic neck mass. AB - Prostate cancer often metastasizes to the regional lymph nodes, but metastases to distant supradiaphragmatic lymph nodes are uncommon. Rare case reports describe cervical lymph node metastases as the first clinical manifestation of prostate cancer, but only in the setting of widely disseminated disease. We present the unusual case of an 84-y-old male with a known history of prostate cancer and recurrent disease limited to the left supraclavicular lymph nodes. PMID- 15897915 TI - Evaluation of a new, rapid, qualitative, one-step PSA Test for prostate cancer screening: the PSA RapidScreen test. AB - To increase the acceptance rate and reduce the cost of the screening programme for prostate cancer, a new qualitative and one-step test for prostate-specific antigen (PSA), called PSA RapidScreen, has been evaluated. PSA RapidScreen test is a chromatographic lateral flow immunoassay, which generates a positive or negative result for PSA values >or=or <4 ng/ml, respectively. Capillary blood samples from 188 men were evaluated. Two independent observers interpreted the test at 10, 15, 20 and 25 min. A total of 10 women were tested as controls. Parallel serum samples were simultaneously collected and tested with an ordinary quantitative assay (Elecsys 2010, Roche). Sensitivity, specificity, accuracy, negative and positive predictive values of the test were 97.6, 90.4, 94, 98 and 89%, respectively. PSA RapidScreen tests on female capillary samples were negative. Reproducibility of the test was 99.5%, while interobserver variation was 5%. Specificity of the test was altered by variations in the reading time. Quantitative assessment of the intensity of the band correlated with the PSA value (r=0.87; t=23.97; P<0.001). PSA RapidScreen is a rapid, simple and reproducible one-step test. The low cost and the speed of the test make it a powerful and convenient tool for prostate cancer screening programmes. PMID- 15897916 TI - Quantitative real-time reverse transcription: polymerase chain reaction of prostate-specific antigen (PSA) for detection of circulating prostatic cells in patients with clinically localized prostate cancer. AB - OBJECTIVE: To evaluate the clinical utility of using the real-time reverse transcriptase-polymerase chain reaction (RT-PCR) to quantify prostate-specific antigen (PSA) mRNA in peripheral blood samples from patients with prostate cancer as a predictor of extraprostatic extension of the disease and to assess any correlations with known predictive markers of this condition. METHODS: Immediately before radical prostatectomy, peripheral blood samples were taken from 42 men with clinically localized prostate cancer and analysed for PSA and 18S ribosomal (endogenous control) genes using real-time RT-PCR (with gene expression assays and the comparative CT-cycle threshold-method for quantifying). A total of 30 healthy male blood donors aged <50 y was taken as a control group. The relationships between PSA mRNA values, pathological and clinical features were analysed. PSA mRNA value, PSA level and biopsy Gleason score were then compared as predictors of extraprostatic extension. RESULTS: PSA gene expression was 3.73 times significantly higher in patients with clinically localized prostate cancer than in healthy men (P<0.05). There was no relationship between PSA real-time RT-PCR values and pathological stage pT2 or pT3 (P=0.5), and no association between PSA mRNA value and serum PSA level (P=0.9) or the Gleason score of the preoperative biopsy (P=0.9). CONCLUSION: There was no significant advantage in using the real-time RT-PCR assay of PSA mRNA before surgery to stage prostate cancer and to discriminate between organ-confined and extraprostatic extension. PMID- 15897917 TI - Subtoxic concentration of doxorubicin enhances TRAIL-induced apoptosis in human prostate cancer cell line LNCaP. AB - Most tumor cells are sensitive to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis but sparing to normal cells, thus providing therapeutic potential for clinical use. Some tumor cells are resistant to TRAIL-induced cell death while the sensitivity could be recruited with the existence of some chemical agents. In this study, human prostatic cancer cell line LNCaP was found to be resistant to TRAIL-induced apoptosis while it could be restored to TRAIL sensitivity with combination treatment of low concentration of doxorubicin. TRAIL receptor-1 (DR4) and TRAIL receptor-2 (DR5) were upregulated under the treatment of doxorubicin and verified to be responsible for TRAIL mediated signal transduction. Furthermore, caspase-8 and caspase-3 were activated and drove their autocleavage into programmed cell death. Interestingly, apoptosis inhibitory protein c-FLIP, but not Bcl-2 and XIAP was downregulated after doxorubicin treatment. Taken together, these findings suggested that the pathway of cell apoptosis induced by TRAIL was intact but under negative control. Subtoxic concentration of doxorubicin effectively boosted TRAIL sensitivity via depletion of antiapoptotic protein. These findings support the new strategies for killing tumors with TRAIL and chemical agents. PMID- 15897918 TI - The role of complement in immunological demyelination of the mammalian spinal cord. AB - STUDY DESIGN: Specificity of serum complement component to elicit immunological demyelination. OBJECTIVES: To assess the role of complement components and pathways in experimental immunological demyelination of the adult rat spinal cord. SETTING: ICORD, University of British Columbia, Vancouver, Canada. SUBJECTS: We used 32 adult male Sprague-Dawley rats, of approximately 220 g weight. METHODS: Rats received intraspinal infusions of demyelinating reagents, delivered by osmotic minipump, for a 7-day infusion at 0.5 microl/h. Reagents consisted of a polyclonal antibody to galactocerebroside and human serum complement. Complement sera deficient for a single component were used to assess the role of the alternative pathway, the classical pathway, and the membrane attack complex. Demyelination was assessed, at 7 days, ultrastructurally. RESULTS: Removal of C3 protein, common to classical and alternative complement pathways, or C4 protein, a classical pathway protein, resulted in no demyelination. However, complement deficient in Factor B, an alternative pathway protein, produced effective demyelination. Upon removal of C5 or C6, membrane attack complex proteins, demyelination was also observed. CONCLUSION: This suggests that the classical pathway is sufficient for the protocol to demyelinate the adult rat spinal cord, and that the membrane attack complex is also not required. PMID- 15897919 TI - Oxidised regenerated cellulose as a cause of paraplegia after thoracotomy: case report and review of the literature. AB - STUDY DESIGN: Case report. OBJECTIVE: To report an unusual case of paraplegia. SETTING: University Hospital Bursa, Turkey. CASE REPORT: A 22-year-old woman presented with paraplegia following a left-sided thoracotomy. Magnetic resonance imaging showed a dorsal epidural mass at the level of T6. The patient underwent an emergency T6/7 laminectomy and removal of a tuft of oxidised regenerated cellulose, which had migrated through the intervertebral foramen causing spinal cord compression. CONCLUSION: In cases of neurological deficits after surgery at the posterolateral edge of a thoracotomy, the clinician should be aware of the above possibility. Urgent radiological diagnosis and decompressive laminectomy is the treatment of choice. PMID- 15897920 TI - Epithelial distribution of neural receptors in the guinea pig small intestine. AB - Neural and paracrine agents, such as dopamine, epinephrine, and histamine, affect intestinal epithelial function, but it is unclear if these agents act on receptors directly at the enterocyte level. The cellular localization and villus crypt distribution of adrenergic, dopamine, and histamine receptors within the intestinal epithelium is obscure and needs to be identified. Single cell populations of villus or crypt epithelial cells were isolated from the jejunum of adult guinea pigs. Enterocytes were separated from intraepithelial lymphocytes by flow cytometry and specific binding was determined using fluorescent probes. Alpha1-adrenergic receptors were located on villus and crypt intraepithelial lymphocytes and enterocytes. Beta-adrenergic receptors were found on villus and crypt enterocytes. Dopamine receptors were found on all cell types examined, whereas histamine receptors were not detected (<10% for each cell population). These studies demonstrated that (1) receptors for epinephrine and dopamine exist on epithelial cells of the guinea pig jejunum, (2) beta-adrenergic receptors are found primarily on villus and crypt enterocytes and (3) intraepithelial lymphocytes contain alpha1-adrenergic, but have few beta-adrenergic, receptors. The presence of neural receptors suggests that these agents are acting, at least in part, at the enterocyte or intraepithelial lymphocyte levels to modulate intestinal and immune function. PMID- 15897921 TI - Involvement of the Na+-independent Cl-/HCO3- exchange (AE) isoform in the compensation of myocardial Na+/H+ isoform 1 hyperactivity in spontaneously hypertensive rats. AB - Enhanced activity of Na+/H+ isoform 1 (NHE-1) and the Na+-independent Cl-/HCO3- exchange (AE) is a feature of the hypertrophied myocardium in spontaneously hypertensive rats (SHR). The present study explored the possibility that sustained intracellular acidosis due to increased myocardial acid loading through AE causes NHE-1 enhancement. To this aim, SHR were treated for 2 weeks with a rabbit polyclonal antibody against an AE3 isoform that was recently developed and proven to have inhibitory effects on myocardial AE activity. We then compared the AE activity in the left ventricle papillary muscles isolated from untreated SHR with antiAE3-treated SHR; AE activity was measured in terms of the rate of intracellular pH recovery after an intracellular alkali load was introduced. AE activity was diminished by approximately 70% in SHR treated with the antiAE3 antibody, suggesting that the AE3 isoform is a major carrier of acid-equivalent influx in the hypertrophied myocardium. However, the antibody treatment failed to normalize NHE-1 activity that remained elevated in the myocardium of normotensive rats. The data therefore rule out the possibility that NHE-1 hyperactivity in hypertensive myocardium was due to sustained intracellular acidosis induced by increased AE activity that characterizes SHR myocardial tissue. PMID- 15897922 TI - Oral administration of sodium tungstate improves cardiac performance in streptozotocin-induced diabetic rats. AB - Normalization of hyperglycemia and hyperlipidemia is an important objective in preventing diabetes-induced cardiac dysfunction. Our study investigated the effects of sodium tungstate on cardiac performance in streptozotocin-induced (STZ) diabetic rats based on its potential antidiabetic and antioxidant activity. Male Wistar rats were made STZ-diabetic and then treated with tungstate in their drinking water for 9 weeks. Body mass, food and fluid intake, plasma glucose, insulin, triglyceride, and free fatty acids levels were measured. At the termination of the study period, an oral glucose tolerance test (OGTT) was performed, and cardiac performance was evaluated using an isolated working heart apparatus. Tungstate-treated STZ-diabetic rats showed a significant reduction in fluid and food intake, plasma glucose, triglycerides, and free fatty acid levels, and improved tolerance to glucose in OGTT, owing to tungstate-mediated enhancement of insulin activity rather than increased insulin levels. Left ventricular pressure development, the rate of contraction (+dP/dT), and the rate of relaxation (-dP/dT) were significantly improved in tungstate-treated diabetic rats. Apart from a decreased rate of body mass gain, no other signs of toxicity or hypoglycemic episodes were observed in tungstate-treated rats. This study extends previous observations on the antidiabetic activities of tungstate, and also reports for the first time the salutary effects in preventing diabetic cardiomyopathy. PMID- 15897923 TI - Sexual dimorphism in angiotensin II-induced hypertension and vascular alterations. AB - Sex differences in the degree of high blood pressure have been described in several forms of experimental animal models of hypertension. However, the influence of sex on angiotensin II-induced hypertension has not been studied. In the present study, we investigated and compared the effects of chronic angiotensin II treatment on blood pressure and vascular function in male and female rats. Chronic treatment with angiotensin II (0.7 mg/kg daily for 10 d) significantly raised arterial blood pressure in male but not female Sprague Dawley rats; it upregulated the NAD(P)H oxidase gp67 phox subunit in the aorta of male but not female rats; and it exaggerated the vasoconstrictor responses to norepinephrine and serotonin in the mesenteric vascular bed (MVB) of male but not female rats. Vasodilator responses to acetylcholine (ACh) but not papaverine (PPV) or isoprenaline (ISO) were reduced in the MVB of angiotensin II-treated male but not female rats. ACh, but not PPV or ISO dilatory responses were potentiated in the MVB of angiotensin II-treated female rats. The present findings demonstrate that exogenous angiotensin II upregulates aortic NAD(P)H oxidase gp67 phox subunit, and induces hypertension and mesenteric vascular dysfunction only in male rats. PMID- 15897924 TI - Reduction of oxidative stress and atherosclerosis in hyperlipidemic rabbits by Dioscorea rhizome. AB - Hyperlipidemia may induce oxidative stress, which is important in the pathogenesis of atherosclerosis. Dioscorea rhizome (DR) is the powdered form of yams, and possesses antioxidant and hypolipidemic function. We therefore investigated the antioxidative and antiatherogenic effects of DR on hyperlipidemic rabbits. The control group was fed chow containing 0.5% cholesterol and 10% corn oil. The probucol and DR groups were fed the same diet as the control group but with the addition of 100 mg probucol/kg chow and 200 mg DR/kg chow, respectively. Total cholesterol and triacylglycerol plasma levels, RBC hemolysis T50, lucigenin chemiluminescence, and luminol chemiluminescence increased in the control group compared with the normal group, and decreased in the probucol and DR groups compared with the control group. The activity of antioxidant enzymes superoxide dismutase and catalase was significantly higher in the probucol and DR group than in the control group. The level of 8-hydroxy-2' deoxyguanosine (8-OHdG) in liver DNA was lower in the probucol and DR group than in the control group. Eighty percent of the intimal surface of the thoracic aorta was covered with atherosclerotic lesions in the control group but only 40% of the surface was covered in the DR group. These results suggest that supplementation with DR reduces oxidative stress and attenuates atherosclerosis in hyperlipidemic rabbits. PMID- 15897925 TI - The effect of glycosaminoglycans with acetaldehyde on the activation of prothrombin. AB - Heparin17-19k, (25, 50, and 100 ng), heparin6k (50 and 100 ng), heparin3k (50, 100, and 200 microg), chondroitin sulfates B (dermatan sulfate) (0.25, 0.5, and 1.0 microg), C (1 and 10 microg), and A (1 and 10 microg) each prolong the activated partial thromboplastin time (APTT) when preincubated with prothrombin to a greater extent than when preincubated with Factor II-deficient plasma prior to their mixing and subsequent additions of APTT reagent and Ca2+. In all cases statistical significance (p < or = 0.05) was observed except with the 2 lower levels of heparin3k. These results suggest that the glycosaminoglycans (GAGs) may exert a direct effect upon prothrombin (FII) in their anticoagulant activity. Pre mix tures of [(FII/25 ng H17-19k) + 447 mmol acetaldehyde (AcH)/L] as well as [(AcH/H) + FII] and [(FII/AcH) + H] each exert a synergistic anticoagulant effect upon APTT. At low AcH concentrations (44.7 mmol/L), neither a synergistic nor an additive effect is seen. H6k and H3k, on premixing with 447 mmol AcH/L, exhibit an additive effect on APTT prolongation but no synergism. Similarly, premixtures of CSB/447 mmol AcH/L/FII show a greater anticoagulant effect than do [(CSB/AcH) + FII] or [(FII/AcH) + CSB] premixtures. CSC-AcH and CSA-AcH patterns are analogous to those of CSB (DS). These data suggest the possibility that AcH, the primary product of ethanol metabolism, may serve as a crosslinking adduct with proteins, in this case, prothrombin, as well as GAGs. Thus ternary complexes between the zymogen form of coagulation factors, GAGs, and AcH are possible, further influencing coagulopathy. PMID- 15897926 TI - Relating drug-induced changes in carotid artery mechanics to cardiovagal and sympathetic baroreflex control. AB - Previous evidence indicates that sensitivity of the baroreflex cardiovagal and sympathetic arms is dissociated. In addition, pharmacologic assessment of baroreflex sensitivity (BRS) has revealed that cardiovagal, but not sympathetic, BRS is greater when blood pressure is increasing versus falling. The origin of this hysteresis is unknown. In this study, carotid artery distensibility and absolute distension (diameter) were assessed to test the hypothesis that vessel mechanics in barosensitive regions affect the BRS of cardiovagal, but not sympathetic, outflow. R-R interval (i.e. time between successive R waves), finger arterial blood pressure, muscle sympathetic nerve activity, and carotid artery dimensions (B-mode imaging) were measured during sequential infusions of sodium nitroprusside (SNP) and phenylephrine (PHE). Systolic and diastolic common carotid artery diameters and pulse pressure were recorded to calculate distensibility of this vessel under each drug condition. Cardiovagal BRS was greater when blood pressure was increasing versus decreasing (p < 0.01). Sympathetic BRS was not affected by direction of pressure change. Distensibility did not differ between SNP and PHE injections. However, compared with SNP, infusion of PHE resulted in larger absolute systolic and diastolic carotid diameters (p < 0.001). Therefore, cardiovagal reflex hysteresis was related to drug-induced changes in common carotid artery diameter but not distensibility. The lack of sympathetic hysteresis in this model suggests a relative insensitivity of this baroreflex component to carotid artery dimensions and provides a possible mechanism for the dissociation between cardiovagal and sympathetic BRS. PMID- 15897927 TI - Pain, sleep disturbance, and fatigue in patients with cancer: using a mediation model to test a symptom cluster. AB - PURPOSE/OBJECTIVES: To test whether sleep disturbance mediates the effect of pain on fatigue. DESIGN: Cross-sectional. SETTING: Radiation therapy clinic, oncology ambulatory clinic, and inpatient oncology unit in an urban teaching hospital. SAMPLE: 84 patients with cancer with multiple primary diagnoses who were experiencing pain. Fifty-three percent were female and 92% were Caucasian, with a mean age of 54 years. METHODS: All participants completed a symptom questionnaire that included the Brief Pain Inventory-Short Form, the Pittsburgh Sleep Quality Index, and the fatigue subscale of the Profile of Mood States questionnaire. Multistage linear regression was used to test a mediation model. MAIN RESEARCH VARIABLES: Fatigue, pain, and sleep disturbance. FINDINGS: Mediation analyses indicate that pain influences fatigue directly as well as indirectly by its effect on sleep. About 20% (adjusted R2 = 0.20) of the variation in fatigue is explained by pain. Thirty-five percent of the variance in fatigue explained by pain was accounted for by the mediation pathway. CONCLUSIONS: Some of the effect of pain on fatigue is mediated by sleep disturbance, but pain has a direct effect on fatigue as well. IMPLICATIONS FOR NURSING: Although the relationship can be explained only partially by the commonsense point of view that people who are in pain lose sleep and naturally report more fatigue, this finding is important and leads to a potential intervention opportunity. Strategies to improve sleep by better pain management may contribute to decreased fatigue. PMID- 15897928 TI - Psychospiritual well-being and symptom distress in women with breast cancer. AB - PURPOSE/OBJECTIVES: To examine the relationship between symptom distress and psychospiritual well-being in women with breast cancer. DESIGN: Descriptive, cross-sectional, correlational study. SETTING: Secondary analysis of data collected in 2000 from the Breast Cancer Support Web site at http://pages.prodigy.net/replyasap/bc. SAMPLE: 100 women were invited to participate in the study after posting an entry in the Web site guest book. Most had stage I or II breast cancer, were nearly 46 years old, and were 10.25 months postdiagnosis. METHODS: Mailed questionnaires. Women were required to meet the following inclusion criteria: a confirmed breast cancer diagnosis, first cancer experience, fewer than two years postsurgery for breast cancer, 18 years of age or older, and the ability to read and write in English. Symptom distress was measured using the Symptom Distress Scale. Psychospiritual well-being was measured by combining scores from the psychological subscale of the breast-cancer specific version of the Functional Assessment of Cancer Therapy Scale and the Functional Assessment of Chronic Illness Therapy (Spiritual) Measurement System 12 for a composite score. MAIN RESEARCH VARIABLES: Symptom distress, psychospiritual well-being. FINDINGS: Symptom distress and psychospiritual well being were inversely related. No relationship was found between age and symptom distress; however, age was inversely related to psychospiritual well-being. Age and symptom distress accounted for 23.1% of the variance in psychospiritual well being. CONCLUSIONS: Participants experienced a small amount of symptom distress, which was inversely related to psychospiritual well-being. Although their symptom distress was similar to other studies, patients in this study reported lower psychospiritual well-being than participants in other studies. IMPLICATIONS FOR NURSING: Psychospiritual well-being is an important concept for nurses seeking a holistic approach to practice because it connects the mind and spirit with the body. PMID- 15897929 TI - Needs of ambulatory patients with cancer who visited outpatient units in Japanese hospitals. AB - PURPOSE/OBJECTIVES: To document the domains and properties of the self-reported needs of ambulatory patients with cancer. DESIGN: Descriptive. SETTING: Outpatient units in three general hospitals in Japan. SAMPLE: 139 ambulatory patients with cancer. METHODS: The data were collected using questionnaires. Five theoretical groups, which were composed of 30 items, were extracted empirically as domains. Alpha coefficients for each domain ranged from 0.70-0.89. Relationships between each domain and other variables and among the domains themselves were examined. MAIN RESEARCH VARIABLES: Expressed needs of ambulatory patients with cancer, their backgrounds, medical and treatment characteristics, and physical functioning. FINDINGS: All domains for patient needs, except for healthcare needs, were negatively correlated with the level of their physical function. Emotional, physical, and functional needs were positively correlated with the frequency of visiting an outpatient unit. Compared with other needs, adaptation needs were greater for patients who were employed or within three months of discharge. Among patients with one of three cancer sites (i.e., breast, stomach, and colorectal cancers), the needs for individualized care were the lowest for patients with colorectal cancer and highest for patients with breast cancer. CONCLUSIONS: From the needs that ambulatory patients with cancer expressed, five domains were derived. Those domains had relationships with other variables. IMPLICATIONS FOR NURSING: The findings shed light on a segment of ambulatory cancer nursing and may be useful when developing and testing programs needed in the future. PMID- 15897931 TI - Depression in men with prostate cancer. AB - PURPOSE/OBJECTIVES: To summarize the current empirical knowledge base on depression in men with prostate cancer to inform psychosocial supportive care interventions for this population and chart directions for future research. DATA SOURCES: Reports in English of quantitative studies that included measures of depression or mood in samples of men with prostate cancer published from 1988 2004. DATA SYNTHESIS: Nurse researchers are playing a key role in establishing the scientific knowledge base upon which a better understanding of the relative importance of depression in men with prostate cancer will emerge. This review indicates that (a) predictable risk factors exist for depression among men with prostate cancer, (b) different prostate cancer treatments do not tend to be associated with differential outcomes in depression or mood, and (c) overall, men with prostate cancer report fewer depressive symptoms than women with breast cancer. CONCLUSIONS: The small body of research addressing depression in men with prostate cancer is methodologically inadequate to estimate the overall prevalence of depression among men with prostate cancer and determine the clinical significance of psychoeducational interventions targeting depression or mood in this population. IMPLICATIONS FOR NURSING: Nurses can use current knowledge to identify men with prostate cancer who are most at risk for depression. Evidence supporting the benefit of psychoeducational interventions for depression in other cancer populations (e.g., women with breast cancer) may be applicable to men with prostate cancer. PMID- 15897932 TI - Information needs of wives of men following prostatectomy. AB - PURPOSE/OBJECTIVES: To identify and evaluate the information needs of wives of men diagnosed with prostate cancer who were treated first with radical prostatectomy and to measure the extent to which their needs were met. DESIGN: Descriptive. SETTING: Comprehensive cancer center designated by the National Cancer Institute. SAMPLE: 66 wives of men with prostate cancer. METHODS: On the second day after their husbands underwent prostatectomies, the wives of men with prostate cancer completed the Family Inventory of Needs-Wives and a demographic data form. MAIN RESEARCH VARIABLES: Information needs and the perceived degree to which they were met. FINDINGS: All needs were identified as important but were met to varying degrees. Five items were not rated "unmet" by any respondents, whereas four items were considered unmet by at least 30% of respondents. Evidence suggests that the age of the wife influences the percentage of unmet needs, with older wives having more unmet needs. P values for the relationship between needs and the age of the wife, her educational level, and the time between diagnosis and treatment were less than 0.05. CONCLUSIONS: Illness prompts a wife's need for information, which supports findings of earlier research. IMPLICATIONS FOR NURSING: Wives have a wide range of information needs. Assessing education needs and the degree to which they are met can guide nurses in restructuring education. Support and guidance regarding more difficult issues, such as fatigue and sexual activity, can decrease barriers to communication. Using a multimodality approach to education, constantly reassessing needs, and encouraging questions are imperative. PMID- 15897933 TI - Prevention of systemic mycoses by reducing exposure to fungal pathogens in hospitalized and ambulatory neutropenic patients. AB - PURPOSE/OBJECTIVES: To describe sources of fungal contamination that can incite invasive mycoses in hospitalized and ambulatory neutropenic patients and to discuss approaches to reduce exposure to pathogens. DATA SOURCES: Published articles, books, and brochures. DATA SYNTHESIS: Modifications of patient environments and lifestyles include hand hygiene for patients and healthcare workers, air filtration in hospitals, and reduction in exposure to plants, soil, standing water, and dusty environments. The effectiveness of dietary restrictions is controversial, although avoidance of pepper is recommended. These restrictions should be implemented prior to, during, and following neutropenia. CONCLUSIONS: Mycoses can be hospital or community acquired; however, although guidelines for environmental and lifestyle modifications are well documented for the institutional setting, they are more limited for ambulatory patients. IMPLICATIONS FOR NURSING: Nurses have a key role in the early identification of outbreaks of fungal infections, evaluation of hospital and home environments for sources of pathogens, education of patients on preventive measures, and research on neutropenic diets and improved technology to reduce exposure to fungal pathogens. PMID- 15897934 TI - Cancer treatment-induced bone loss in patients with breast or prostate cancer. AB - PURPOSE/OBJECTIVES: To review the prevalence, consequences, pathophysiology, diagnosis, and treatment of cancer treatment-induced bone loss (CTIBL) in patients with breast or prostate cancer. DATA SOURCES: Published articles, abstracts, book chapters, electronic resources, and manufacturer information. DATA SYNTHESIS: CTIBL is a long-term complication associated with cancer therapies that cause hypogonadism in patients with breast or prostate cancer. Early diagnosis and treatment of CTIBL is essential to prevent bone fractures. CTIBL treatment includes modification of lifestyles that increase the risk of developing bone loss and fractures and includes the use of bone loss therapies (e.g., bisphosphonates, selective estrogen receptor modifiers, calcitonin). CONCLUSIONS: CTIBL is becoming more common as patients with breast or prostate cancer survive longer. Identifying and treating CTIBL early are important because once bone is lost, damaged bone becomes more difficult to repair; early diagnosis and treatment also may prevent fractures. IMPLICATIONS FOR NURSING: Nurses must be knowledgeable about CTIBL to identify high-risk patients and educate patients and their families about CTIBL, bone loss therapies, and lifestyle modifications. PMID- 15897935 TI - Strengths and limitations of breast cancer risk assessment. AB - PURPOSE/OBJECTIVES: To evaluate current definitions of breast cancer risk and breast cancer risk assessment models, including the Gail, Claus, and BRCAPRO models, and discuss potential markers to enhance and standardize individual risk assessment. DATA SOURCES: Published articles, conference proceedings, and textbooks. DATA SYNTHESIS: Defining high risk for breast cancer development is explored, and options for high-risk women are discussed. The risk factors frequently used for risk evaluation, including age, age at menarche, age at first live birth, past history of breast biopsy, family history of breast cancer, and the presence of atypical hyperplasia, are reviewed. CONCLUSIONS: Current models of breast cancer risk assessment are limited. Exploring the progression from healthy tissue to malignancy through techniques such as fine needle aspiration, ductal lavage, and nipple aspiration may lead to more precise individualized risk prediction. IMPLICATIONS FOR NURSING: More accurate information regarding personal breast cancer risk is necessary. Oncology nurses may facilitate the use of appropriate tools that provide the most individualized risk assessment. PMID- 15897936 TI - Do women in the community recognize hereditary and sporadic breast cancer risk factors? AB - PURPOSE/OBJECTIVES: To describe knowledge of hereditary, familial, and sporadic breast cancer risk factors among women in the community and to identify characteristics associated with this knowledge. DESIGN: Descriptive, cross sectional. SETTING: Community settings in the San Francisco Bay Area. SAMPLE: 184 women who had never been diagnosed with cancer, were 30-85 years old (mean = 47 + 12), and agreed to complete a questionnaire in English. Participants were from diverse racial and cultural backgrounds (i.e., 43% European descent, 27% African descent, 16% Asian descent, and 14% Hispanic descent). Many (49%) were college graduates, and 24% had a median annual family income of $30,000-$50,000. METHODS: Survey. MAIN RESEARCH VARIABLES: Knowledge of hereditary, familial, and sporadic breast cancer risk factors and characteristics associated with this knowledge. FINDINGS: Although most women recognized heredity as a risk factor, some did not understand the impact of paternal family history on risk. Some women did not recognize the relationship between breast and ovarian cancer, risk factors associated with the Gail model, and that aging increases risk. Education level was the most important characteristic associated with knowledge of risk factors. CONCLUSIONS: Although age and family history are independent predictors of sporadic, hereditary, and familial breast cancer risk, women in the community could not distinguish between the three forms of the disease. Although the sample included a large number of educated women, their knowledge of breast cancer risk factors appeared incomplete. IMPLICATIONS FOR NURSING: Advanced practice nurses should provide individualized risk assessment and education regarding breast cancer risk factors. PMID- 15897937 TI - How do nurse case managers care for older women with breast cancer? AB - PURPOSE/OBJECTIVES: To describe how nurse case managers care for older women with breast cancer. DESIGN: A randomized, prospective trial. SETTING: Thirteen community hospitals and two public hospitals in southeastern Texas. SAMPLE: 335 older women aged 60-89 years newly diagnosed with breast cancer and being cared for by 60 surgeons. Most participants were Caucasian. A total of 166 women were in the control group, and 159 were in the intervention nurse case management group. METHODS: The nurses implemented multiple nursing interventions in each nursing process phase over a period of 12 months. MAIN RESEARCH VARIABLES: Independent variables were participants' demographic characteristics, depressive symptomatology, and cognitive impairment. Nurse case management contact was a dependent variable. FINDINGS: In each nursing phase, a greater number of nurse case management contacts were made in the first quarter. Bivariate analysis illustrated statistical differences among race, income, education, and living alone with respect to the mean amount of nurse case management. Multivariate analysis revealed that age, income, living alone, and stage of cancer predicted more nurse case management contact. CONCLUSIONS: Nurse case managers may play a role in helping older women with breast cancer achieve positive health outcomes. IMPLICATIONS FOR NURSING: Based on the findings of this study, nurses can develop specific nursing interventions to meet the needs of older women with breast cancer. Nurses can use the Model of Nurse Case Management to plan and manage care for older women with breast cancer. PMID- 15897938 TI - Fatigue, psychological distress, coping resources, and functional status during radiotherapy for uterine cancer. AB - PURPOSE/OBJECTIVES: To evaluate how patients diagnosed with uterine cancer experience fatigue, psychological distress, coping resources, and functional status before, during, and after treatment with radiation therapy and to study whether significant correlations exist among these variables. DESIGN: Longitudinal, descriptive, and correlational. SETTING: The Department of Oncology, Sahlgrenska University Hospital, Gothenburg, Sweden. SAMPLE: 60 women diagnosed with uterine cancer who were receiving curative external radiation therapy. Typical participants were 64 years old, married, and on sick leave or retired from work. METHODS: Data were collected through self-report instruments. Demographic and clinical data were extracted from the patients' records. MAIN RESEARCH VARIABLES: Cancer-related fatigue, psychological distress, coping resources, and functional status. FINDINGS: Patients' fatigue scores increased significantly during and after completion of radiotherapy. The participants reported normal levels of anxiety and depression, and their coping resources changed over time. After completing therapy, all dimensions of function had decreased; for social function, the decrease was significant. The correlation over time was significant among fatigue and physical function, role function, and cognitive function. The variation of the change in fatigue after therapy was completed was explained only by the level of fatigue experienced at baseline. CONCLUSIONS: Fatigue is a symptom that increases in connection with radiotherapy. Functional status is influenced by the variation in fatigue levels. Fatigue level before treatment may be an important variable when trying to find a risk factor for the development of fatigue over the course of treatment. IMPLICATIONS FOR NURSING: Nurses must inform patients receiving radiotherapy about the expected changes in fatigue and functional status. Pretreatment screening for fatigue is needed to identify patients at risk for developing fatigue. PMID- 15897939 TI - A phenomenologic study of fatigue in adolescents receiving treatment for cancer. AB - PURPOSE/OBJECTIVES: To generate a detailed description of how adolescents with cancer manage their daily lives and the way in which fatigue affects this. DESIGN: Phenomenologic. SETTING: A pediatric oncology unit at a regional cancer center in the United Kingdom. SAMPLE: A convenience sample of adolescents (N = 8), aged 16-19 years and with hematologic or solid tumors, who currently were undergoing primary treatment. METHODS: Semistructured interviews were conducted using 11 open-ended questions. MAIN RESEARCH VARIABLES: Adolescents' perceptions of fatigue, well-being, and ability to maintain normal activities. FINDINGS: Adolescents reported fatigue as overwhelming and embedded in a syndrome of symptoms and emotions associated with the illness itself and with treatment. Fatigue had a significant effect on physical, psychological, and social well being, placing an extra burden on adolescents who were striving for normality. CONCLUSIONS: Equipped with a rich description of fatigue, clinicians will be better prepared to initiate strategies congruent with their own work settings and particular patients. IMPLICATIONS FOR NURSING: The findings should enable healthcare professionals to construct a more accurate and perceptive picture of the needs of particular individuals, highlighting those that may be amenable to intervention. PMID- 15897940 TI - Nutritional, functional, and emotional characteristics related to fatigue in patients during and after biochemotherapy. AB - PURPOSE/OBJECTIVES: To test Winningham's psychobiologic entropy hypothesis in patients receiving biochemotherapy for melanoma. DESIGN: Descriptive, correlational, cross-sectional study. SETTING: Midwest cancer center. SAMPLE: 25 male and female patients who were receiving biochemotherapy or who had completed treatment 6-12 months prior. METHODS: Data were collected using a series of questionnaires and diet recall. MAIN RESEARCH VARIABLES: Fatigue, anxiety, depression, distressing symptoms, nutritional intake, and weight. FINDINGS: Moderate fatigue was significantly related to physiologic and psychological symptoms but not to nutrient intake. The sample was overweight, and a significant number of participants were obese. High caloric intakes were evident. Depression was a significant problem. CONCLUSIONS: Fatigue was not as severe as expected, but problems with responses to the fatigue scale may explain this. Nutritional status and nutrient intake were not correlated to fatigue in this sample. Activity levels were related to fatigue, and treatment reduced activity. On average, activity returned to pretreatment levels 6-12 months after treatment. Winningham's hypothesis held and will be useful for understanding fatigue in this population. IMPLICATIONS FOR NURSING: Depression needs to be assessed and treated as a side effect of biotherapy. Assessing the impact of nutrition when patients are overweight or obese is difficult. A scale specifically designed to test Winningham's hypothesis is needed. PMID- 15897941 TI - Content validity of self-report measurement instruments: an illustration from the development of the Brain Tumor Module of the M.D. Anderson Symptom Inventory. AB - PURPOSE/OBJECTIVES: To illustrate one technique for establishing content validity of measurements using the initial development and testing of the M.D. Anderson Symptom Inventory Brain Tumor Module. DATA SOURCES: Published articles, book chapters, and subjective judgments of experts. DATA SYNTHESIS: Content validity is the essential first step in the development of items to be included in a measurement instrument. Content validity is a criterion-referenced process that is judged by how well each item in a newly developed instrument reflects its respective objective or content domain. The stages in addressing content validity include a developmental stage and a judgment-quantification stage. Steps involved in the developmental stage include domain identification, item generation, and instrument formation. The judgment-quantification stage is when experts review the items and either report validity of the items subjectively or with an empirically referenced method, such as calculation of the content validity index. The content validity of a set of questions designed to measure symptoms in a population of patients with primary brain tumors was ascertained by using the calculation of the content validity index. CONCLUSIONS: The final version of the M.D. Anderson Symptom Inventory Brain Tumor Module consists of the 13 core items and 18 additional items designated as valid by a panel of experts. The instrument will be administered to a group of patients to determine construct validity and reliability of the items. IMPLICATIONS FOR NURSING: Self-report instruments are used to measure various health outcomes in oncology. Oncology nurses are in a key position to develop such instruments to be used in clinical care and research of symptoms associated with cancer. Understanding the process of content validation is an essential first step in developing new instruments. PMID- 15897942 TI - Perceptions of gender roles, gender power relationships, and sexuality in Thai women following diagnosis and treatment for cervical cancer. AB - PURPOSE/OBJECTIVES: To describe patients' and their partners' perceptions of gender roles, gender power relationships, and sexuality before diagnosis of and after treatment for cervical cancer. DESIGN: Descriptive. SETTING: Southern Thailand. SAMPLE: 97 women with cervical cancer who received cancer treatment, including radiotherapy, and their partners. METHODS: Structured interview methods were used to gather information relating to gender roles, gender power relationships, and sexuality. MAIN RESEARCH VARIABLES: Gender roles, gender power relationships, and sexuality. FINDINGS: Fifty-two percent of the women (n = 50) were diagnosed with stage II cervical cancer. The percentage of women who undertook various activities specific to gender roles before their diagnoses was higher than the percentage who undertook the same activities after treatment. An increased percentage of partners undertook the women's gender-role-specific activities after the women received cancer treatment compared with the percentage who did so before diagnosis. Little change in gender power relationships was reported. A high percentage of the couples reported changes in various aspects of their sexuality after cancer treatment compared with before diagnosis. CONCLUSIONS: Gender roles, gender power relationships, and sexuality changed for women with cervical cancer and their partners after the women completed cancer treatment. IMPLICATIONS FOR NURSING: Open discussions among women with cervical cancer, their partners, and oncology nurses are necessary to identify culturally sensitive and appropriate solutions. PMID- 15897943 TI - Neuroimaging: anything to do with neurotherapeutics? PMID- 15897945 TI - Efficacy of proton magnetic resonance spectroscopy in neurological diagnosis and neurotherapeutic decision making. AB - Anatomic and functional neuroimaging with magnetic resonance imaging (MRI) includes the technology more widely known as magnetic resonance spectroscopy (MRS). Now a routine automated "add-on" to all clinical magnetic resonance scanners, MRS, which assays regional neurochemical health and disease, is therefore the most accessible diagnostic tool for clinical management of neurometabolic disorders. Furthermore, the noninvasive nature of this technique makes it an ideal tool for therapeutic monitoring of disease and neurotherapeutic decision making. Among the more than 100 brain disorders that fall within this broad category, MRS contributes decisively to clinical decision making in a smaller but growing number. In this review, we will cover how MRS provides therapeutic impact in brain tumors, metabolic disorders such as adrenoleukodystrophy and Canavan's disease, Alzheimer's disease, hypoxia, secondary to trauma or ischemia, human immunodeficiency virus dementia and lesions, as well as systemic disease such as hepatic and renal failure. Together, these eight indications for MRS apply to a majority of all cases seen. This review, which examines the role of MRS in enhancing routine neurological practice and treatment concludes: 1) there is added value from MRS where MRI is positive; 2) there is unique decision-making information in MRS when MRI is negative; and 3) MRS usefully informs decision making in neurotherapeutics. Additional efficacy studies could extend the range of this capability. PMID- 15897944 TI - Foundations of advanced magnetic resonance imaging. AB - During the past decade, major breakthroughs in magnetic resonance imaging (MRI) quality were made by means of quantum leaps in scanner hardware and pulse sequences. Some advanced MRI techniques have truly revolutionized the detection of disease states and MRI can now-within a few minutes-acquire important quantitative information noninvasively from an individual in any plane or volume at comparatively high resolution. This article provides an overview of the most common advanced MRI methods including diffusion MRI, perfusion MRI, functional MRI, and the strengths and weaknesses of MRI at high magnetic field strengths. PMID- 15897946 TI - Molecular optical imaging: applications leading to the development of present day therapeutics. AB - A number of advances in the molecular imaging field have led to the sensing of specific molecular targets and pathways in living animals. In the optical imaging field, these include the designing of biocompatible near-infrared fluorochromes, development of targeted and activatable "smart" imaging probes, and engineering of activatable fluorescent and bioluminescent proteins. The current advances in molecular optical imaging will help in early disease diagnoses, functioning of a number of pathways and finally help speed drug discovery. In this review, we will describe the near infrared fluorescent and bioluminescence imaging modalities and how these techniques have been employed in current research. Furthermore, we will also shed some light on the use of these imaging modalities in neurotherapeutics, for example imaging different parameters of vector-mediated gene expression in glioma tumors and stem cell tracking in vivo. PMID- 15897947 TI - Positron emission tomography and single-photon emission computed tomography in central nervous system drug development. AB - In this review, the value of functional imaging [positron emission tomography (PET)/single-photon emission computed tomography (SPECT)] in drug development is considered. Radionuclide imaging can help establish the diagnosis of neurodegenerative disorders where this is in doubt and provides a potential biomarker for following drug effects on disease progression. PET and SPECT can help understand mechanisms of disease and determine the functional effects of therapeutic approaches on neurotransmission and metabolism. Synthesizing radiotracer analogs of novel drugs can provide proof of principle that these agents reach their enzyme or receptor targets and delineate their regional brain distribution. If such radiotracers do not prove to have ideal properties for imaging, the concept of microdosing potentially allows multiple other drug analogs to be tested with less stringent regulatory requirements than for novel medicinals. Finally, PET tracers can provide receptor and enzyme active site dose occupancy profiles, thereby guiding dosage selection for phase 1 and phase 2 trials. The eventual hope is that radiotracer imaging will provide a surrogate marker for drug efficacy, although this has yet to be realized, and progress the concept of personalized medicine where receptor/enzyme binding profiles help predict therapeutic outcome. PMID- 15897948 TI - Single-photon emission computed tomography in neurotherapeutics. AB - The measurement of regional cerebral blood flow (rCBF) by single-photon emission computed tomography (SPECT) is a powerful clinical and research tool. There are several clinical applications now documented, a substantial number under active investigation, and a larger number yet to be studied. Standards regarding patient imaging environment and image presentation are becoming established. This article reviews key aspects of SPECT functional brain imaging in clinical practice, with a particular emphasis on therapeutics, including 1) the quality of the tomographic device, 2) the radiopharmaceutical employed, 3) environmental conditions at the time of radiotracer administration, 4) characteristics of the subject, 5) the format used for image presentation, and 6) the essential components of image processing necessary to the achievement of high-quality SPECT brain images. Next, a brief description of relevant radiation safety issues is provided. Finally, applications in molecular imaging, especially in small animal imaging for research as well as drug discovery and development are discussed. The gamut of SPECT studies from currently routine clinical applications to molecular imaging offers a wonderful frontier for opportunities to employ functional brain imaging in neurotherapeutics. PMID- 15897949 TI - Magnetic resonance imaging, microscopy, and spectroscopy of the central nervous system in experimental animals. AB - Over the last two decades, microscopic resolution in vivo magnetic resonance imaging (MRI) techniques have been developed and extensively used in the study of animal models of human diseases. Standard MRI methods are frequently used in clinical studies and in the general clinical practice of human neurological diseases. This generates a need for similar studies in experimental animal research. Because small rodents are the most commonly used species as animal models of neurological diseases, the MRI techniques need to be able to provide microscopic resolution and high signal-to-noise ratio images in relatively short time. Small animal MRI systems use very high field-strength magnets, which results in higher signal to noise ratio; however, the contrast characteristics of live tissue are different at these field strengths. In addition to standard MRI techniques, several new applications have been implemented in experimental animals, including diffusion and perfusion studies, MR angiography, functional MRI studies, MRI tractography, proton and phosphorous spectroscopy, cellular and molecular imaging using novel contrast methods. Here we give an overview of how to establish a small animal imaging facility with the goal of CNS imaging. We describe the basic physical processes leading to MR signal generation, highlighting the differences between standard clinical MRI and small animal MRI. Finally, typical findings in the most common neurological disease categories and novel MRI/magnetic resonance spectroscopy methods used in their study are also described. PMID- 15897950 TI - Neuroimaging of ischemia and infarction. AB - Since the introduction of thrombolytic therapy as the foundation of acute stroke treatment, neuroimaging has rapidly advanced to empower therapeutic decision making. Diffusion-weighted imaging is the most sensitive and accurate method for stroke detection, and, allied with perfusion-weighted imaging, provides information on the functional status of the ischemic brain. It can also help to identify a response to thrombolytic and neuroprotective therapies. Additionally, multimodal magnetic resonance imaging, including magnetic resonance angiography, offers information on stroke mechanism and pathophysiology that can guide long term medical management. Multimodal computed tomography is a comprehensive, cost effective, and safe stroke imaging modality that can be easily implemented in the emergency ward and that offers fast and reliable information with respect to the arterial and functional status of the ischemic brain. Accessibility, contraindications, cost, speed, and individual patient-determined features influence which is the best imaging modality to guide acute stroke management. PMID- 15897951 TI - Imaging of multiple sclerosis: role in neurotherapeutics. AB - Magnetic resonance imaging (MRI) plays an ever-expanding role in the evaluation of multiple sclerosis (MS). This includes its sensitivity for the diagnosis of the disease and its role in identifying patients at high risk for conversion to MS after a first presentation with selected clinically isolated syndromes. In addition, MRI is a key tool in providing primary therapeutic outcome measures for phase I/II trials and secondary outcome measures in phase III trials. The utility of MRI stems from its sensitivity to longitudinal changes including those in overt lesions and, with advanced MRI techniques, in areas affected by diffuse occult disease (the so-called normal-appearing brain tissue). However, all current MRI methodology suffers from limited specificity for the underlying histopathology. Conventional MRI techniques, including lesion detection and measurement of atrophy from T1- or T2-weighted images, have been the mainstay for monitoring disease activity in clinical trials, in which the use of gadolinium with T1-weighted images adds additional sensitivity and specificity for areas of acute inflammation. Advanced imaging methods including magnetization transfer, fluid attenuated inversion recovery, diffusion, magnetic resonance spectroscopy, functional MRI, and nuclear imaging techniques have added to our understanding of the pathogenesis of MS and may provide methods to monitor therapies more sensitively in the future. However, these advanced methods are limited by their cost, availability, complexity, and lack of validation. In this article, we review the role of conventional and advanced imaging techniques with an emphasis on neurotherapeutics. PMID- 15897952 TI - Therapeutic advances in interventional neurology. AB - Rapid advances in the field of interventional neurology and the development of minimally invasive techniques have resulted in a great expansion of potential therapeutic applications. We discuss therapeutic interventional neurology as applied in clinical practice in one of the two possible ways: 1) embolization leading to occlusion of blood vessels; and 2) revascularization leading to reopening of blood vessels. These procedures can be applied to a broad range of cerebrovascular diseases. In the first section of this review, we will explore the evolution of these interventions to occlude aneurysms, arteriovenous malformations, neurovascular tumors, and injuries. In the second section, revascularization in acute ischemic stroke, stenosis, and dural venous thrombosis will be discussed. PMID- 15897953 TI - Neuroimaging of infections. AB - Neuroimaging plays a crucial role in the diagnosis and therapeutic decision making in infectious diseases of the nervous system. The review summarizes imaging findings and recent advances in the diagnosis of pyogenic brain abscess, ventriculitis, viral disease including exotic and emergent viruses, and opportunistic disease. For each condition, the ensuing therapeutic steps are presented. In cases of uncomplicated meningitis, cranial computed tomography (CT) appears to be sufficient for clinical management to exclude acute brain edema, hydrocephalus, and pathology of the base of skull. Magnetic resonance imaging (MRI) is superior in depicting complications like sub-/epidural empyema and vasculitic complications notably on FLAIR (fluid-attenuated inversion recovery) weighted images. The newer technique of diffusion-weighted imaging (DWI) shows early parenchymal complications of meningitis earlier and with more clarity and is of help in differentiation of pyogenic abscess (PA) from ring enhancing lesions of other etiology. Proton magnetic resonance spectroscopy (PMRS) seems to produce specific peak patterns in cases of abscess. The presence of lactate cytosolic amino acids and absence of choline seems to indicate PA. Also in cases of suspected opportunistic infection due to toxoplasma DWI may be of help in the differentiation from lymphoma, showing no restriction of water diffusion. In patients with herpes simplex and more exotic viruses like West Nile and Murray Valley virus DWI allows earlier lesion detection and therapeutic intervention with virustatic drugs. PMID- 15897954 TI - Imaging in neurooncology. AB - Imaging in patients with brain tumors aims toward the determination of the localization, extend, type, and malignancy of the tumor. Imaging is being used for primary diagnosis, planning of treatment including placement of stereotaxic biopsy, resection, radiation, guided application of experimental therapeutics, and delineation of tumor from functionally important neuronal tissue. After treatment, imaging is being used to quantify the treatment response and the extent of residual tumor. At follow-up, imaging helps to determine tumor progression and to differentiate recurrent tumor growth from treatment-induced tissue changes, such as radiation necrosis. A variety of complementary imaging methods are currently being used to obtain all the information necessary to achieve the above mentioned goals. Computed tomography and magnetic resonance imaging (MRI) reveal mostly anatomical information on the tumor, whereas magnetic resonance spectroscopy and positron emission tomography (PET) give important information on the metabolic state and molecular events within the tumor. Functional MRI and functional PET, in combination with electrophysiological methods like transcranial magnetic stimulation, are being used to delineate functionally important neuronal tissue, which has to be preserved from treatment induced damage, as well as to gather information on tumor-induced brain plasticity. In addition, optical imaging devices have been implemented in the past few years for the development of new therapeutics, especially in experimental glioma models. In summary, imaging in patients with brain tumors plays a central role in the management of the disease and in the development of improved imaging-guided therapies. PMID- 15897955 TI - Neuroimaging biomarkers for clinical trials of disease-modifying therapies in Alzheimer's disease. AB - The pathophysiologic process leading to neurodegeneration in Alzheimer's disease (AD) is thought to begin long before clinical symptoms develop. Existing therapeutics for AD improve symptoms, but increasing efforts are being directed toward the development of therapies to impede the pathologic progression of the disease. Although these medications must ultimately demonstrate efficacy in slowing clinical decline, there is a critical need for biomarkers that will indicate whether a candidate disease-modifying therapeutic agent is actually altering the underlying degenerative process. A number of in vivo neuroimaging techniques, which can reliably and noninvasively assess aspects of neuroanatomy, chemistry, physiology, and pathology, hold promise as biomarkers. These neuroimaging measures appear to relate closely to neuropathological and clinical data, such as rate of cognitive decline and risk of future decline. As this work has matured, it has become clear that neuroimaging measures may serve a variety of potential roles in clinical trials of candidate neurotherapeutic agents for AD, depending in part on the question of interest and phase of drug development. In this article, we review data related to the range of neuroimaging biomarkers of Alzheimer's disease and consider potential applications of these techniques to clinical trials, particularly with respect to the monitoring of disease progression in trials of disease-modifying therapies. PMID- 15897956 TI - Neuroimaging and therapeutics in movement disorders. AB - In this review, we discuss the role of neuroimaging in assessing treatment options for movement disorders, particularly Parkinson's disease (PD). Imaging methods to assess dopaminergic function have recently been applied in trials of potential neuroprotective agents. Other imaging methods using regional metabolism and/or cerebral perfusion have been recently introduced to quantify the modulation of network activity as an objective marker of the treatment response. Both imaging strategies have provided novel insights into the mechanisms underlying a variety of pharmacological and stereotaxic surgical treatment strategies for PD and other movement disorders. PMID- 15897957 TI - Neuroimaging in traumatic brain imaging. AB - Traumatic brain injury (TBI) is a common and potentially devastating clinical problem. Because prompt proper management of TBI sequelae can significantly alter the clinical course especially within 48 h of the injury, neuroimaging techniques have become an important part of the diagnostic work up of such patients. In the acute setting, these imaging studies can determine the presence and extent of injury and guide surgical planning and minimally invasive interventions. Neuroimaging also can be important in the chronic therapy of TBI, identifying chronic sequelae, determining prognosis, and guiding rehabilitation. PMID- 15897959 TI - Antifreeze glycoproteins from the antarctic fish Dissostichus mawsoni studied by differential scanning calorimetry (DSC) in combination with nanolitre osmometry. AB - This study investigates in detail the freezing events during cooling of solutions of various size classes of antifreeze glycoproteins. Differential scanning calorimetry and nanolitre osmometry were used to observe ice growth at temperatures within the hysteresis gap between the melting point and non equilibrium freezing point (hysteresis freezing point) of solutions of the various sized antifreeze glycoproteins (AFGPs). The ice growth within the hysteresis gap is presumably due to both the expression of primary or near primary prism planes and also some growth at the basal plane. The binding of the AFGPs to the ice causes a particular ice crystal morphology. With the smaller AFGPs, substantial microscopic ice growth was observed in the form of a hexagonal bipyramids within the hysteresis gap. PMID- 15897958 TI - Neuroimaging of epilepsy: therapeutic implications. AB - Neuroimaging has important applications in the diagnosis and treatment of patients with seizures and epilepsy. Having replaced computed tomography (CT) in many situations, MRI is the preferred imaging technique for patients with epilepsy. Advances in radionuclide-based techniques such as single-photon emission CT/positron emission tomography and electromagnetic source imaging with magnetoencephalography are providing new insights into the pathophysiology of epilepsy. In addition, techniques such as magnetic resonance spectroscopy are beginning to impact treatment. In this review, I discuss how these techniques are used in clinical practice but more importantly, how imaging findings play an increasing role in neurotherapeutics. PMID- 15897960 TI - Effect of three vitrification methods on development of two-cell mouse embryos. AB - The purpose of this study was to investigate the effect of three vitrification procedures [conventional straw (CS), open pulled straw (OPS) and closed pulled straw (CPS)] on the development of two-cell mouse embryos. Two-cell mouse embryos were randomly divided into vitrified and non-vitrified control groups. Embryos in the vitrified group were cryopreserved within a combination of 5.5 M ethylene glycol and 1M sucrose as cryoprotectants, loaded within three different straws (CS, OPS and CPS) and warmed in stepwise sucrose solutions. The survived embryos from each procedure were cultured in human tubal fluid (HTF). The non-vitrified control embryos were also cultured in the same manner. The rates of the development in all the groups were daily determined and statistically compared. On day 4 of the cultivation period, several expanded blastocysts from each group were randomly selected and stained either with propidium iodide (PI) and bisbenzimide or with terminal transferase- mediate DNA end labeling (TUNEL) Technique. The mean number of the inner cell mass (ICM), trophoectoderm (TE), necrotic and apoptotic cells were counted and statistically compared. The survival rate of embryos in CPS was significantly higher than that in OPS and CS. The rate of hatched blastocysts did not differ in the three vitrification procedures, but in comparison with that of the control, CS and OPS showed a significant reduction. The mean number of ICM and TE decreased in CS and OPS, whereas in CPS it was almost identical to that of the control. The incidence of apoptosis and necrosis appeared to be almost similar in all the groups. In conclusion, CPS seems to be an effective, easy and rapid method for the cryopreservation of two-cell mouse embryos. PMID- 15897961 TI - Cryopreservation of yams using vitrification modified by including droplet method: effects of cold acclimation and sucrose. AB - Cryopreservation of yams (Dioscorea spp.) is important for the preservation of valuable genotypes for food, medicine and breeding purposes. This study on four species of yams was conducted to evaluate the influence of cold acclimation in an alternating 5 degree C and 28 degree C, 12 h thermo-photo-period and of two sucrose concentrations in the preculture medium using a modified droplet method. Acclimation of a 3-week period provided the best preconditioning treatment averaged over four genotypes. Effect of sucrose concentration in the preculture medium depended on the genotype (significant genotype x sucrose interaction; P = 0.036). High survival (67 to 70%) with 30% to 50% shoot recovery was obtained for D. bulbifera, D. polystachya and D. cayenensis, compared to 20 percent survival without shoot recovery for D. alata. PMID- 15897962 TI - Changes in sucrose and glycerol content in garlic shoot tips during freezing using PVS3 solution. AB - Changes in moisture content (MC), sucrose and glycerol concentration in garlic shoot tips were monitored during loading and unloading with PVS3 solution. Upon PVS3 treatment, shoot tip MC decreased rapidly and sucrose and glycerol concentrations increased rapidly during the first 30 min. Sucrose and glycerol concentrations increased more slowly thereafter. Shoot tip MC in after PVS3 treatment was affected by their size, but not by sucrose concentration of the preculture medium. As the size of shoot tips increased, so their MC increased after PVS3 treatment. However, sucrose and glycerol concentrations decreased after PVS3 incubation, and concentrations in dehydrated shoot tips were much lower than those measured in non-air dried controls. During unloading with 1.2 M sucrose medium, shoot tip MC increased rapidly during the first 10 min, whereas glycerol concentration decreased steadily over 90 min. Loading and unloading of PVS3 solution in garlic shoot tips follows the principle of solute bulk flow. PMID- 15897963 TI - Thermal stress study of two different artery cryopreservation methods. AB - Two methods used in artery deep cryopreservation, "Cryopreservation in Medium" and "Cryopreservation in Air", were studied. For the former method, samples were frozen together with a certain amount of cryoprotectants (CPA) in the cryovial or cryobag, while for the other method the arteries were first exposed to CPA and then frozen without the CPA medium surrounding in the cryovial. Study of the cryopreserved arteries using these two methods found that "cryopreservation in air" could substantially reduce the fracture rate of the arteries. To explain the difference theoretically, a two-compartment model is presented to study the thermal stresses generated during the freezing and thawing processes. The properties were measured as inputs to the model. Numerical results showed that the thermal stresses occurring in the "cryopreservation in air" process were much smaller than in the other method. The maximum thermal stress during cryopreservation occurs in the thawing process. The theoretical results could well explain published experimental results. PMID- 15897964 TI - Survival of four accessions of Anigozanthos manglesii (haemodoraceae) seeds following exposure to liquid nitrogen. AB - This study investigated the survival of seeds from the prominent endemic Western Australian species Anigozanthos manglesii following exposure to liquid nitrogen (cryostorage). Seeds from four different accessions (collected in 1987, 1990, 1993 and 1998) adjusted to different water contents were tested for survival following cryostorage. Water content was a significant determining factor with survival of cryostored seeds declining rapidly at water contents above c. 18%. These water contents were deemed as critical water contents and were supported by DSC scans showing high endothermic peaks indicating ice crystallisation. In some instances, survival of cryostored seeds also declined at low water contents. Seeds from 1990 had a lower than expected survival compared to the other accessions. This may have resulted from the higher lipid content of seeds from this accession, or the reduced germination and vigour of these seeds prior to cryostorage. PMID- 15897965 TI - Integrity of endothelium in cryopreserved human cornea. AB - The aim of the present study was to elaborate an optimal method for cryopreservation of human donor cornea for transplantation and to follow the morphological changes in the structure of the endothelial cell layer using scanning electron microscopy (SEM). Sixteen groups, with four donor cornea each, were cryopreserved at cooling rates of 1 degree C per min and 5 degree C per min. Four cryoprotectants (glycerol, dimethyl sulfoxide, 1,2-propanediol, polyethylene glycol-400) in two concentrations (5% and 10% v/v) were prepared on the bases of medium Optisol GS supplied with 20% v/v human serum albumin. Four additional human cornea were used as controls. Endothelial cell recovery of the cornea after thawing and 24 hours culture, was calculated as a percent of the preserved recovered cells. Sufficient recovery of the endothelial cell layer, making the cornea suitable for transplantation was obtained using the cryoprotectants dimethyl sulfoxide and especially polyethylene glycol-400. PMID- 15897966 TI - RNA in human sperm. AB - We have yet to develop a fundamental understanding of the molecular complexities of human spermatozoa. This encompasses the unique packaging and structure of the sperm genome along with their paternally derived RNAs in preparation for their delivery to the egg. The diversity of these transcripts is vast, including several anti-sense molecules resembling known regulatory micro-RNAs. The field is still grasping with its delivery to the oocyte at fertilization and possible significance. It remains tempting to analogize them to maternally-derived transcripts active in early embryo patterning. Irrespective of their role in the embryo, their use as a means to assess male factor infertility is promising. PMID- 15897967 TI - Novel association between sperm deformity index and oxidative stress-induced DNA damage in infertile male patients. AB - AIM: To investigate the impact of abnormal sperm morphology using the sperm deformity index (SDI) on reactive oxygen species (ROS) production and its correlation with sperm DNA damage. METHODS: Semen samples were collected from men undergoing infertility screening (n = 7) and healthy donors (n = 6). Mature spermatozoa were isolated and incubated with 5 mmol/L beta-nicotinamide adenine dinucleotide phosphate (NADPH) for up to 24 h to induce ROS. Sperm morphology was evaluated using strict Tygerberg's criteria and the SDI. ROS levels and DNA damage were assessed using chemiluminescence and terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling (TUNEL) assays, respectively. RESULTS: SDI values (median [interquartiles]) were higher in patients than donors (2 [1.8, 2.1] vs. 1.53 [1.52, 1.58], P = 0.008). Aliquots treated with NADPH showed higher ROS levels (1.22 [0.30, 1.87] vs. 0.39 [0.10, 0.57], P = 0.03) and higher incidence of DNA damage than those not treated (10 [4.69, 24.85] vs. 3.85 [2.58, 5.10], P = 0.008). Higher DNA damage was also seen following 24 h of incubation in patients compared to donors. SDI correlated with the percentage increase in sperm DNA damage following incubation for 24 h in samples treated with NADPH (r = 0.7, P = 0.008) and controls (r = 0.58, P = 0.04). CONCLUSION: SDI may be a useful tool in identifying potential infertile males with abnormal prevalence of oxidative stress (OS)-induced DNA damage. NADPH plays a role in ROS-mediated sperm DNA damage, which appears to be more evident in infertile patients with semen samples containing a high incidence of morphologically abnormal spermatozoa. PMID- 15897968 TI - Identification of a novel testis-specific gene and its potential roles in testis development/spermatogenesis. AB - AIM: To identify and characterize a novel gene with potential roles in testis development and spermatogenesis. METHODS: A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes. Differentially expressed genes were isolated and sequenced. RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis. A range of bioinformatical programs including Gene Runner, SMART, NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature. RESULTS: A novel testis-specific gene, NYD-SP5, was differentially expressed in fetal and adult testes. The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved Ile and Gln residues), a Carbamate kinase-like domain, a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain. RT PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined. In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes. CONCLUSION: NYD-SP5 is a newly found testis-specific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme. PMID- 15897969 TI - Cryopreservation of cynomolgus monkey (Macaca fascicularis) spermatozoa in a chemically defined extender. AB - AIM: To establish a method for cynomolgus monkey sperm cryopreservation in a chemically defined extender. METHODS: Semen samples were collected by electro ejaculation from four sexually mature male cynomolgus monkeys. The spermatozoa were frozen in straws by liquid nitrogen vapor using egg-yolk-free Tes-Tris mTTE synthetic extender and glycerol as cryoprotectant. The effects of glycerol concentration (1 %, 3 %, 5 %, 10 % and 15 % [v/v]) and its equilibration time (10 min, 30 min, 60 min and 90 min) on post-thaw spermatozoa were examined by sperm motility and sperm head membrane integrity. RESULTS: The post-thaw motility and head membrane integrity of spermatozoa were significantly higher (P0.05) for 5 % glycerol (42.95 +/- 2.55 and 50.39+/- 2.42, respectively) than those of the other groups (1%: 19.19 +/- 3.22 and 24.84 +/- 3.64; 3%: 34.23 +/- 3.43 and 41.37 +/- 3.42; 10%:15.68 +/- 2.36 and 21.39 +/- 3.14; 15%: 7.47 +/- 1.44 and 12.90 +/- 2.18). The parameters for 30 min equilibration(42.95 +/- 2.55 and 50.39 +/- 2.42) were better (P0.05) than those of the other groups (10 min: 31.33 +/- 3.06 and 38.98 +/- 3.31; 60 min: 32.49 +/- 3.86 and 40.01 +/- 4.18; 90 min: 31.16 +/- 3.66 and 38.30 +/- 3.78). Five percent glycerol and 30 min equilibration yielded the highest post-thaw sperm motility and head membrane integrity. CONCLUSION: Cynomolgus monkey spermatozoa can be successfully cryopreserved in a chemically defined extender, which is related to the concentration and the equilibration time of glycerol. PMID- 15897971 TI - Saposin C stimulates growth and invasion, activates p42/44 and SAPK/JNK signaling pathways of MAPK and upregulates uPA/uPAR expression in prostate cancer and stromal cells. AB - AIM: To determine the effect of saposin C (a known trophic domain of prosaposin) on proliferation, migration and invasion, as well as its effect on the expression of urokinase plasmonogen activator (uPA), its receptor (uPAR) and matrix metalloproteinases (MMP)-2 and -9 in normal and malignant prostate cells. In addition, we tested whether saposin C can activate p42/44 and stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK) signal transduction pathways of the mitogen-activated protein kinase (MAPK) superfamily. METHODS: We employed Western blot analysis, phospho-specific antibodies, cell proliferation assay, reverse transcriptase-polymerase chain reaction, in vitro kinase assays and migration and invasion to determine the effect of saposin C on various biological behaviors of prostate stromal and cancer cells. RESULTS: Saposin C, in a cell type-specific manner, upregulates uPA/uPAR and immediate early gene c-Jun expression, stimulates cell proliferation, migration and invasion and activates p42/44 and SAPK/JNK MAPK pathways in prostate stromal and cancer cells. Normal prostate epithelial cells were not responsive to saposin C treatment in the above studies. CONCLUSION: Saposin C functions as a multipotential modulator of diverse biological activities in prostate cancer and stromal cells. These results strongly suggest that saposin C functions as a potent growth factor for prostatic cells and may contribute to prostate carcinogenesis and/or the development of hormone-refractory prostate cancer. PMID- 15897972 TI - Mass screening of prostate cancer in a Chinese population: the relationship between pathological features of prostate cancer and serum prostate specific antigen. AB - AIM: To investigate the pathological features of the prostate biopsy through mass screening for prostate cancer in a Chinese cohort and their association with serum prostate specific antigen (PSA). METHODS: A total of 12027 Chinese men in Changchun were screened for prostate cancer by means of the serum total prostate specific antigen tPSA test (by Elisa assay). Transrectal ultrasound-guided systematic six-sextant biopsies were performed on those whose serum tPSA value was > 4.0 ng/mL and those who had obstructive symptoms (despite their tPSA value) and were subject to subsequent pathological analysis with the aid of the statistic software SPSS 10.0 (SPSS. Inc., Chicago. USA). RESULTS: Of the 12027 cases, 158 (including 137 patients whose serum tPSA values were 4.0 ng/mL and 21 patients [serum tPSA < 4.0 ng/mL] who had obstructive symptoms) undertook prostate biopsy. Of the 158 biopsies, 41 cases of prostatic carcinoma were found (25.9 %, 41/158). The moderately differentiated carcinoma and poorly differentiated carcinoma accounted for 61% and 34%, respectively. A significant linear positive correlation between the serum tPSA and the Gleason scores in the 41 cases of prostatic carcinoma (r = 0.312, P < 0.01) was established. A significant linear positive correlation between the serum tPSA value of the 41 prostatic carcinoma and the positive counts of carcinoma in sextant biopsies was established (r = 0.406, P < 0.01), indicating a significant linear relationship between serum tPSA and the size of tumor. CONCLUSION: This study was the first to conduct mass screening for prostate cancer by testing for serum tPSA values and the first to investigate the pathological features of prostate cancer in a cohort of Chinese men. Our results reveal that the moderately differentiated carcinoma is the most common type of prostate cancer. This study also has shown that the serum tPSA value in prostate cancer is associated with the Gleason score and the size of tumor. PMID- 15897973 TI - Ultrasound-guided transrectal extended prostate biopsy: a prospective study. AB - AIM: To evaluate the diagnostic value of the 10 systematic transrectal ultrasound guided (TRUS) prostate biopsy compared with the sextant biopsy technique for patients with suspected prostate cancer. METHODS: One hundred and fifty-two patients with suspected prostate cancer were included in the study. Patients were entered in the study because they presented with high levels of prostate specific antigen (PSA) (over 4 ng/mL) and/or had undergone an abnormal digital rectal examination (DRE). In addition to sextant prostate biopsy cores, four more biopsies were obtained from the lateral peripheral zone with additional cores from each suspicious area revealed by transrectal ultrasound. Sextant, lateral peripheral zone and suspicious area biopsy cores were submitted separately to the pathological department. RESULTS: Cancer detection rates were 27.6% (42/152) and 19.7% (30/152) for the 10-core and sextant core biopsy protocols, respectively. Adding the lateral peripheral zone (PZ) to the sextant prostate biopsy showed a 28.6% (12/42) increase in the cancer detection rate in patients with positive prostate cancer (P < 0.01). The cancer detection rate in patients who presented with elevated PSA was 29.3% (34/116). When serum PSA was 4-10 ng/mL TRUS-guided biopsy detected cancer in 20.6%, while the detection rate was 32.4% and 47.0% when serum PSA was 10-20 ng/mL and above 20 ng/mL, respectively. CONCLUSION: The 10 systematic TRUS-guided prostate biopsy improves the detection rate of prostate cancer by 28.6% when compared with the sextant biopsy technique alone, without increase in the morbidity. We therefore recommend the 10-core biopsy protocol to be the preferred method for early detection of prostate cancer. PMID- 15897974 TI - Novel functional association of rat testicular membrane-associated cytosolic glutathione S transferases and cyclooxygenase in vitro. AB - AIM: To analyze the role of cytosolic glutathione S-transferases cGSTs and membrane-associated cytosolic GSTs macGSTs in prostaglandin biosynthesis and to evaluate the possible interaction between glutathione S-transferases GSTs and cyclooxygenase (COX) in vitro. METHODS: SDS-PAGE analysis was undertaken for characterization of GSTs, thin layer chromatography (TLC) to monitor the effect of GSTs on prostaglandin biosynthesis from arachidonic acid (AA) and spectrophotometric assays were done for measuring activity levels of COX and GSTs. RESULTS: SDS-PAGE analysis indicates that macGSTs have molecular weights in the range of 25-28 kDa. In a coupled assay involving GSTs, arachidonic acid and cyclooxygenase-1, rat testicular macGSTs produced prostaglandin E2 and F2alfa, while the cGSTs caused the generation of prostaglandin D2, E2 and F2alfa. In vitro interaction studies on GSTs and COX at the protein level have shown dose dependent inhibition of COX activity by macGSTs and vice versa. This effect, however, is not seen with cGSTs. The inhibitory effect of COX on macGST activity was relieved with increasing concentrations of reduced glutathione (GSH) but not with 1-chloro 2,4-dinitrobenzene (CDNB). The inhibition of COX by macGSTs, on the other hand, was potentiated by glutathione. CONCLUSION: We isolated and purified macGSTs and cGSTs from rat testis and analyzed their involvement in prostaglandin biosynthesis. These studies reveal a reversible functional interaction between macGSTs and COX in vitro, with possible interactions between them at the GSH binding site of macGSTs. PMID- 15897975 TI - Expression of a novel beta adaptin subunit mRNA splice variant in human testes. AB - AIM: To identify a novel isoform of adaptin 2 beta subunit (named Ap2beta-NY) and to investigate its relationship with testicular development and spermatogenesis. METHODS: Using a human testis cDNA microarray, a clone (Ap2beta-NY), which was strongly expressed in adult testes but weakly expressed in embryo testes, was sequenced and analyzed. Using polymerase chain reaction (PCR), the tissue distribution and expression time pattern of Ap2beta-NY were determined. RESULTS: Ap2beta-NY was identified and has been deposited in the GenBank (AY341427). The expression level of Ap2beta-NY in the adult testis was about 3-fold higher than that in the embryo testis. PCR analysis using multi-tissue cDNA indicated that Ap2beta-NY was highly expressed in the testis, spleen, thymus, prostate, ovary, blood leukocyte and brain, but not in the heart, placenta, lung, liver, skeletal muscle, kidney and pancreas. In addition, Ap2beta-NY was variably expressed in the testes of patients with spermatogenesis-disturbance and spermatogenesis arrest but not expressed in those of Sertoli-cell-only syndrome, which implied that, in the testis, Ap2beta-NY was restrictively expressed in germ cells. CONCLUSION: Ap2beta-NY is an isoform of Ap2beta and may be involved in regulating the process of spermatogenesis and testis development. PMID- 15897976 TI - Evaluation of spermatogenesis and fertility in F1 male rats after in utero and neonatal exposure to extremely low frequency electromagnetic fields. AB - AIM: To determine whether in utero and neonatal exposure to a 60 Hz extremely low frequency electromagnetic field (EMF) results in spermatotoxicity and reproductive dysfunction in the F1 offspring of rats. METHODS: Age-matched, pregnant Sprague-Dawley rats were exposed continuously (21 h/day) to a 60 Hz EMF at field strengths of 0 (sham control), 5, 83.3 or 500 microT from day 6 of gestation through to day 21 of lactation. The experimentally generated magnetic field was monitored continuously (uninterrupted monitoring over the period of the study) throughout the study. RESULTS: No exposure-related changes were found in exposed or sham-exposed animals with respect to the anogenital distance, preputial separation, testis weight, testicular histology, sperm count, daily sperm production, sperm motility, sperm morphology and reproductive capacity of F1 offspring. CONCLUSION: Exposure of Sprague-Dawley rats to a 60 Hz EMF at field strengths of up to 500 microT from day 6 of gestation to day 21 of lactation did not produce any detectable alterations in offspring spermatogenesis and fertility. PMID- 15897977 TI - Safety evaluation of long-term vas occlusion with styrene maleic anhydride and its non-invasive reversal on accessory reproductive organs in langurs. AB - AIM: To evaluate the safety of the long term vas occlusion with styrene maleic anhydride (SMA) and its non-invasive reversal at the level of accessory reproductive glands ARGs in langurs. METHODS: The morphology of seminal vesicle and ventral prostate was evaluated by light as well as transmission electron microscopy. Serum clinical chemistry and urine albumin were evaluated in an autoanalyzer using reagent kits. Fructose, acid phosphatase and zinc in the seminal plasma were evaluated spectrophotometrically according to the WHO manual. Serum testosterone, prostate specific antigen and sperm antibodies were evaluated by enzyme-linked immunosorbent assays (ELISA) using reagent kits and hematology was estimated according to standard procedures. RESULTS: The morphological features and secretory activity of the seminal vesicle and prostate were normal as evidenced by the presence of well-developed mitochondria, rough endoplasmic reticulum, Golgi bodies, secretory granules and normal nuclear characteristics throughout the course of investigation. Serum testosterone and prostate specific antigen remained unaltered and serum antisperm antibodies level presented negative titres. Urine albumin was nil. Total red blood corpuscles (RBC), white blood corpuscles (WBC), hemoglobin (Hb) and red cell indices, serum protein, glucose, cholesterol, creatinine, creatine kinase (CK), serum glutamate oxalate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), lactate dehydrogenase (LDH), bilirubin, urea, triglycerides and high-density lipoprotein (HDL) did not show appreciable changes following vas occlusion and after its non invasive reversal. Although fructose, acid phosphatase (ACP) and zinc in the seminal plasma showed a significant reduction following vas occlusion, it could not be related to the morphology of seminal vesicle and prostate. CONCLUSION: SMA vas occlusion and its non-invasive reversal do not damage the accessory reproductive organs. PMID- 15897978 TI - Identification and characterization of cul-3b, a novel hominine CUL-3 transcript variant. AB - AIM: To identify genes related to the human testis development by substrate hybridization technique. METHODS: A human testis cDNA microarray was constructed and hybridized with probes prepared from human adult and fetal testes and spermatozoa mRNAs by reverse transcription reactions. The differentially expressed genes were sequenced. And a newly identified cullin-3 (CUL-3) transcript variant (designated cul-3b) was bio-informatically analyzed with an online GenBank database. Multi-tissue reverse transcription polymerase chain reaction (RT-PCR) was used to determine the tissue expression profile of cul-3b. RESULTS: Cul-3b, a novel CUL-3 transcript variant, was identified. The expression level of cul-3b in adult testes was 3.79-fold higher than that in fetal ones. Cul 3b differed from cul-3 (including NM_003590 and AY337761) in the opening reading frame and had three internal ribosomal entry sites IRESes in the 5'-UTR. These led to a 24 amino acid (aa) truncation at N-terminus of CUL-3b as compared with CUL-3 and a more motivated expression pattern of cul-3b under some strict circumstances. Additionally, cul-3b expressed ubiquitously in human tissues according to multi-tissue RT-PCR. CONCLUSION: Cul-3b is a novel transcript variant of CUL-3, which may be important not only for the development of human testis but also for that of other organs. PMID- 15897979 TI - Y microdeletions in the Istria county, Croatia. AB - AIM: To establish the frequency of Y chromosome microdeletions in an unselected group of infertile Croatian men. METHODS: An unselected group of 105 patients (male partners of infertile couples), both with idiopathic and non-idiopathic infertility, consecutively referred to the outpatient infertility clinic, gynecology department, General Hospital Pula, Istria County, Croatia, was examined for the presence or absence of Y chromosome microdeletions by polymerase chain reaction analysis. RESULTS: One of the 105 men (0.95%, 95% CI = 0.17-5.2%) was found to have a microdeletion. CONCLUSION: A low frequency of Y chromosome microdeletions was found in the group of unselected infertile Croatian men. PMID- 15897980 TI - Birth after intracytoplasmic sperm injection of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome. AB - AIM: To report a birth after intracytoplasmic sperm injection (ICSI) of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome detected by fluorescence in situ hybridization (FISH) analysis. METHODS: A 35-year-old man with a normal appearance consulted our hospital because of sterility over a 5 year period. Chromosome analysis showed low-incidence mosaic Klinefelter's syndrome. Using FISH, 96 % hyperploidy of the lymphocytes was found. We examined the sex chromosome of the ejaculated spermatozoa. Using FISH, we examined 200 ejaculated spermatozoa and no hyperploidy was found. RESULTS: The 33-year-old female partner of the male patient underwent an uncomplicated controlled ovarian hyperstimulation sequence using a combined recombinant-follicle stimulating hormone (rec-FSH) + human menopausal gonadotrophin (hMG) protocol, following late luteal phase pituitary down regulation. This culminated in the retrieval of seven oocytes, six of which were fertilized with ICSI. One ICSI attempt led to clinical pregnancy with a healthy baby girl. CONCLUSION: We report a male patient with low incidence mosaic Klinefelter's syndrome whose ejaculated spermatozoa were identified as being haploid by FISH before ICSI, leading to the successful pregnancy of his wife and the birth of a healthy baby girl. PMID- 15897982 TI - Polymorphisms within the C-reactive protein (CRP) promoter region are associated with plasma CRP levels. AB - Elevated plasma levels of C-reactive protein (CRP), an inflammation-sensitive marker, have emerged as an important predictor of future cardiovascular disease and metabolic abnormalities in apparently healthy men and women. Here, we performed a systematic survey of common nucleotide variation across the genomic region encompassing the CRP gene locus. Of the common single-nucleotide polymorphisms (SNPs) identified, several in the CRP promoter region are strongly associated with CRP levels in a large cohort study of cardiovascular risk in European American and African American young adults. We also demonstrate the functional importance of these SNPs in vitro. PMID- 15897983 TI - Changing dynamics of varicella-zoster virus infections in the 21st century: the impact of vaccination. PMID- 15897984 TI - Incidence of herpes zoster, before and after varicella-vaccination-associated decreases in the incidence of varicella, 1992-2002. AB - BACKGROUND: Varicella zoster virus (VZV) causes varicella and, later in the life of the host, may reactivate to cause herpes zoster (HZ). Because it is hypothesized that exposure to varicella may boost immunity to latent VZV, the vaccination-associated decrease in varicella disease has led some to suggest that the incidence of HZ might increase. We assessed the impact that varicella vaccination has on the incidence of varicella and of HZ. METHODS: Codes for cases of varicella and of HZ in an HMO were determined in automated databases of inpatients and outpatients, on the basis of the Ninth Revision of the International Classification of Diseases. We calculated the incidence, during 1992-2002, of varicella and of HZ. RESULTS: The incidence of HZ remained stable as the incidence of varicella decreased. Age-adjusted and -specific annual incidence rates of varicella decreased steadily, starting with 1999. The age adjusted rates decreased from 2.63 cases/1000 person-years during 1995 to 0.92 cases/1000 person-years during 2002; among children 1-4 years old, there was a 75% decrease between 1992-1996 and 2002. Age-adjusted and -specific annual incidence rates of HZ fluctuated slightly over time; the age-adjusted rate was highest, at 4.05 cases/1000 person-years, in 1992, and was 3.71 cases/1000 person years in 2002. CONCLUSIONS: Our findings revealed that the vaccination-associated decrease in varicella disease did not result in an increase in the incidence of HZ. These early findings will have to be confirmed as the incidence of varicella disease continues to decrease. PMID- 15897985 TI - No serological evidence of association between prostate cancer and infection with herpes simplex virus type 2 or human herpesvirus type 8: a nested case-control study. AB - Sexual history has consistently been found to be a risk factor for the development of prostate cancer. An association between prostate cancer and herpes simplex virus type 2 (HSV-2) or Kaposi sarcoma-associated herpesvirus/human herpesvirus type 8 (HHV-8) infections has also been reported. Linkage of data on a cohort of 20,243 healthy Finnish men identified 165 cases of prostate cancer that were diagnosed up to 24 years after donation of a serum sample. Two control subjects were matched by age, sex, and municipality of residence to each case patient. Serum levels of immunoglobulin G against HSV-2 and HHV-8 were determined. Neither HSV-2 infection (odds ratio [OR], 0.93 [95% confidence interval {CI}, 0.44-1.96]) nor HHV-8 infection (OR, 0.74 [95% CI, 0.19-2.88]) was associated with prostate cancer. PMID- 15897986 TI - Longitudinal analysis of herpes simplex virus-specific CD4+ cell clonotypes in infected tissues and blood. AB - BACKGROUND: Genital infection by herpes simplex virus (HSV)-2 offers a unique model for study of the effects of a remitting and exacerbating infection on the survival and persistence of antigen-specific T cells. METHODS: We used complementarity-determining region 3 (CDR3) length analysis to examine the complete T cell receptor (TCR) beta -chain repertoire in skin-lesion biopsy samples from subjects with genital herpes. RESULTS: We found that herpetic skin lesions consistently demonstrated oligoclonal CDR3 DNA length distribution, indicating the presence of T cell expansions. Sequence analysis of representative HSV-specific lesional CD4(+) cell clones and TCR beta -variable (TCRBV) sequencing confirmed that the oligoclonal expansions were largely related to HSV specific T cell proliferation. To assess the persistence of HSV-specific CD4(+) cells that localize to genital lesions, we developed a sensitive and highly specific clonal tracking technique using a combination of TCRBV-specific polymerase chain reaction, followed by liquid hybridization with clonotype specific probes. CONCLUSION: Two different patterns of clonal persistence were observed. Some long-lasting clones appear to home to different epithelia, such as skin and genital mucosa, and to circulate in the peripheral blood, whereas others detected in lesions were absent or very rare in the peripheral blood. PMID- 15897987 TI - Epidemiological and virological characteristics of 2 subgroups of hepatitis B virus genotype C. AB - BACKGROUND: We aimed to investigate the characteristics of hepatitis B virus (HBV) genotype C subgroups in Hong Kong and their relationship with HBV genotype C in other parts of Asia. METHODS: Full-genome nucleotide sequences of 49 HBV genotype C isolates from Chinese patients with chronic hepatitis B were compared with the sequences of 69 HBV genotype C isolates and 12 non-genotype C isolates in the GenBank database. Phylogenetic analysis was performed to define the subgroups of HBV genotype C on the basis of >4% heterogeneity of the entire HBV genome. RESULTS: HBV in 80% of patients in Hong Kong belonged to a subgroup predominantly found in Southeast Asia (Vietnam, Thailand, Myanmar, and southern China) designated as HBV genotype "Cs," and HBV in the remaining 20% of patients belonged to another subgroup, predominantly found in the Far East (Korea, Japan, and northern China), designated as HBV genotype "Ce." Overall, the mean+/-SD nucleotide sequence difference between HBV genotype Cs and HBV genotype Ce was 4.2%+/-0.3%. When HBV genotype Cs and HBV genotype Ce were compared among patients in Hong Kong, HBV genotype Cs was associated with a higher tendency to develop basal core promoter mutations (80% vs. 50%; P=.14), a higher prevalence of C at nucleotide 1858 (95% vs. 0%; P<.001), and a lower prevalence of precore stop codon mutations (5% vs. 50%; P=.002). CONCLUSIONS: HBV genotype C can be differentiated into 2 subgroups--namely, genotype Ce and genotype Cs--that have different epidemiological distributions and virological characteristics. PMID- 15897988 TI - Antigenic cross-reactivity between severe acute respiratory syndrome-associated coronavirus and human coronaviruses 229E and OC43. AB - Cross-reactivity between antibodies to different human coronaviruses (HCoVs) has not been systematically studied. By use of Western blot analysis, indirect immunofluorescence assay (IFA), and enzyme-linked immunosorbent assay (ELISA), antigenic cross-reactivity between severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV) and 2 HCoVs (229E and OC43) was demonstrated in immunized animals and human serum. In 5 of 11 and 10 of 11 patients with SARS, paired serum samples showed a > or =4-fold increase in antibody titers against HCoV-229E and HCoV-OC43, respectively, by IFA. Overall, serum samples from convalescent patients who had SARS had a 1-way cross-reactivity with the 2 known HCoVs. Antigens of SARS-CoV and HCoV-OC43 were more cross-reactive than were those of SARS-CoV and HCoV-229E. PMID- 15897989 TI - Declining incidence of invasive Streptococcus pneumoniae infections among persons with AIDS in an era of highly active antiretroviral therapy, 1995-2000. AB - BACKGROUND: Our goal was to describe trends in invasive pneumococcal disease incidence among persons with acquired immunodeficiency syndrome (AIDS) since the introduction of highly active antiretroviral therapy (HAART). METHODS: We used time-trend analysis of annual invasive pneumococcal disease incidence rates from a population-based, active surveillance system. Annual incidence rates were calculated for 5 July-June periods by use of data from San Francisco county, the 6-county Baltimore metropolitan area, and Connecticut. The numerators were the numbers of invasive Streptococcus pneumoniae infections among persons 18-64 years of age with AIDS; the denominators were the numbers of persons living with AIDS, estimated on the basis of AIDS surveillance data. RESULTS: The annual incidence of invasive pneumococcal disease declined from 1094 cases/100,000 persons with AIDS (July 1995-June 1996) to 467 cases/100,000 persons living with AIDS (July 1999-June 2000). The annual percentage changes in incidence were -34%, -29%, -8%, and -1%. Declines were similar by surveillance area, sex, and race/ethnicity. During the final year of the study, the invasive pneumococcal disease incidence in persons with AIDS was half that of the pre-HAART era but was still 35 times higher than that in similarly aged non-HIV-infected adults. CONCLUSIONS: In the United States, invasive pneumococcal disease incidence declined sharply across a range of subgroups living with AIDS during the period after widespread introduction of HAART. Despite these gains, persons with AIDS remain at high risk for invasive pneumococcal disease. PMID- 15897990 TI - Relationship between adherence and the development of resistance in antiretroviral-naive, HIV-1-infected patients receiving lopinavir/ritonavir or nelfinavir. AB - BACKGROUND: Relationships between adherence to protease inhibitor (PI)-based therapy and resistance development have not been fully characterized. METHODS: We conducted a double-blind, randomized, controlled study of lopinavir/ritonavir versus nelfinavir, each administered with stavudine and lamivudine, in 653 antiretroviral-naive, human immunodeficiency virus (HIV)-1-infected patients. Relationships between adherence and probability of resistance development were evaluated by local linear regression or logistic regression. RESULTS: A higher risk of detectable HIV-1 RNA loads after week 24 was associated with lower adherence (odds ratio [OR], 1.08 per 1% decrease in adherence [95% confidence interval {CI}, 1.05-1.10]; P<.001) and nelfinavir use (OR, 2.4 vs. lopinavir/ritonavir [95% CI, 1.6-3.6]; P<.001). Among all nelfinavir-treated patients, a bell-shaped relationship between adherence and the risk of nelfinavir resistance was observed, with a maximum probability of 20% at 85%-90% adherence. No lopinavir resistance was observed. A bell-shaped relationship was also observed for the probability of lamivudine resistance, with a maximum probability of 50% at 75%-80% adherence to nelfinavir and of 15% at 80%-85% adherence to lopinavir/ritonavir. CONCLUSIONS: Bell-shaped relationships between adherence and resistance were observed. Irrespective of adherence level, the risk of detectable HIV-1 RNA loads or of PI or lamivudine resistance was significantly higher in nelfinavir-treated patients than in lopinavir/ritonavir-treated patients. PMID- 15897991 TI - Combination of HIV-1-specific CD4 Th1 cell responses and IgG2 antibodies is the best predictor for persistence of long-term nonprogression. AB - BACKGROUND: Strong T cell and antibody responses to human immunodeficiency virus (HIV), low virus production, and some genetic traits have been individually associated with nonprogression of HIV infection, but the best correlate with protection against disease progression remains unknown. METHODS: We prospectively followed 66 untreated long-term nonprogressors and analyzed relationships between HIV-1-specific CD4 T helper (Th) 1 and CD8 T cell responses and HIV-1-specific antibodies, HIV-1 RNA and proviral DNA loads, host genes, and CD4 Th1 cell counts at entry into the study and 4 years later. RESULTS: HIV-1 p24-specific CD4 Th1 cell proliferation, interferon (IFN)- gamma production, and IFN- gamma -producing cell frequencies at entry significantly and negatively correlated with HIV-1 RNA and proviral DNA loads and were independent of CD4 Th1 cell counts and host genes. HIV-1 Gag-specific IFN- gamma -producing CD8 T cell frequencies correlated with HIV-1 proviral DNA loads but not with RNA loads. Only high frequencies of HIV-1 p24-specific CD4 Th1 cells combined with HIV-1 gp41-specific IgG2 antibodies significantly predicted persistence of high CD4 Th1 cell counts. CONCLUSION: HIV-1-specific CD4 Th1 responses combined with IgG2 antibodies and IFN- gamma -producing CD4 Th1 cells are better predictors of long-term nonprogression than are virus parameters, host genes, or HIV-1-specific CD4 Th1 or CD8 T cell proliferation. PMID- 15897992 TI - Functional genomic relationships in HIV-1 disease revealed by gene-expression profiling of primary human peripheral blood mononuclear cells. AB - BACKGROUND: An assessment of biomarkers from an analysis of human peripheral blood mononuclear cell gene-expression profiles was made, to acquire an understanding of transcriptional changes associated with human immunodeficiency virus type 1 (HIV-1) infection in vivo. METHODS: Supervised learning algorithms were used to create signature gene sets that could be used to distinguish seropositive from seronegative samples and delineate changes in disease status during the early stages of infection. Bioinformatic tools were used to classify persons and to functionally characterize groups of differentially expressed genes, to elucidate the impact of viral infection on host cell gene-expression patterns. RESULTS: A 10-gene signature set that could be used to accurately determine the HIV-1 serostatus was identified. A 6-gene signature set was used to distinguish seropositive persons exhibiting differential changes in CD4(+) T cell counts, with 93% accuracy. Functional classification of differentially expressed genes in HIV-1 indicated a preponderance of down-regulated genes with functions related to the immune response and apoptosis. Hierarchical cluster analysis in persons whose CD4(+) T cell counts increased, compared with that in persons whose CD4(+) T cell counts decreased, was characterized by the down-regulation of genes associated with apoptosis, mitochondrial function, protein biosynthesis, and RNA binding. CONCLUSIONS: Gene-expression profile analysis of a complex infectious virus, such as HIV-1, may be useful to elucidate the functional genomic relationships associated with viral infection. PMID- 15897993 TI - Helicobacter pylori, pepsinogen, and gastric adenocarcinoma in Hawaii. AB - BACKGROUND: The objective was to investigate the association of Helicobacter pylori and serum pepsinogen (PG) levels with gastric adenocarcinoma. METHODS: Serum obtained from 299 patients at the time of cancer diagnosis and from 336 population-based control subjects was tested for PG I, PG II, and antibodies to H. pylori and to CagA. RESULTS: Subjects with low PG I levels or low PG I/II ratios were at increased risk for cardia and noncardia gastric cancer, whereas those with H. pylori or CagA seropositivity had an elevated risk for noncardia cancer only. Subjects seropositive for either H. pylori or CagA who had low PG I levels had the highest odds ratio (OR) (9.21 [95% confidence interval {CI}, 4.95 17.13]) for noncardia cancer, compared with subjects with neither factor. Elevated risks were also found among subjects with only 1 factor (OR, 5.40 [95% CI, 2.61-11.20] for low PG I level only; OR, 4.86 [95% CI, 5.90-8.13] for H. pylori or CagA seropositivity only). This pattern persisted when PG I/II ratio replaced PG I level and when CagA seropositivity alone replaced H. pylori immunoglobulin G or CagA seropositivity. CONCLUSIONS: The results suggest that persons with both H. pylori or CagA seropositivity and a low PG I level or PG I/II ratio are highly susceptible to development of noncardia gastric cancer. PMID- 15897994 TI - Mechanisms underlying Campylobacter fetus pathogenesis in humans: surface-layer protein variation in relapsing infections. AB - Campylobacter fetus causes gastrointestinal and systemic infections in humans. Although relapse is common despite antibiotic treatment, the mechanisms are not well understood. The surface-layer proteins (SLPs) of C. fetus, which are critical in virulence, undergo high-frequency phenotypic switching due to recombination of sap homologues, resulting in antigenic variation. To investigate the mechanisms involved in relapsing C. fetus infections, we compared SLP variation in 4 pairs of C. fetus strains that infect humans; initial and follow up isolations were performed 20 days to 34 months apart. Of the 4 pairs of strains, 2 had antigenic variation, and another provided evidence for selection for SLP-positive populations. Southern hybridization indicated recombination underlying the SLP variation and up-regulation. The fourth pair had the same SLP antigenic profile and sap homologue hybridization pattern, which is consistent with latency of the original strain in a privileged locus. In total, these findings indicate that relapse may reflect at least 3 differing pathogenetic pathways. PMID- 15897995 TI - Angiotensin II subtype 1 receptor blockade inhibits Clostridium difficile toxin A induced intestinal secretion in a rabbit model. AB - Angiotensin II (ANG II) has been described in the regulation of intestinal secretion and absorption via angiotensin subtype 1 (AT(1)) and AT(2) receptors, respectively, in rats. We investigated the role that ANG II plays in the rabbit ileal-loop model of Clostridium difficile infection. Expression of AT(1), the more abundant ANG II receptor, was demonstrated in ileal loops, and an AT(1) receptor blocker, losartan, inhibited hypersecretion induced by C. difficile toxin A (mean volume : length ratio, 0.27+/-0.06 vs. 0.60+/-0.06 mL/cm in controls). Losartan also decreased production of ANG II in the ileum (0.48+/-0.06 vs. 0.87+/-0.12 pg/mg in controls), raising the possibility that ANG II may participate in a positive feedback loop involving the hypersecretory response. Our findings suggest that ANG II plays a significant role in the pathogenesis of C. difficile toxin-induced diarrhea. PMID- 15897996 TI - Identification of the optimal structure required for a Shiga toxin neutralizer with oriented carbohydrates to function in the circulation. AB - Shiga toxin (Stx) is a major virulence factor of Stx-producing Escherichia coli. Recently, we developed a therapeutic Stx neutralizer with 6 trisaccharides of globotriaosyl ceramide, a receptor for Stx, in its dendrimer structure (referred to as "SUPER TWIG [1]6") to function in the circulation. Here, we determined the optimal structure of SUPER TWIG for it to function in the circulation and identified a SUPER TWIG with 18 trisaccharides, SUPER TWIG (2)18, as another potent Stx neutralizer. SUPER TWIGs (1)6 and (2)18 shared a structural similarity, a dumbbell shape in which 2 clusters of trisaccharides were connected via a linkage with a hydrophobic chain. The dumbbell shape was found to be required for formation of a complex with Stx that enables efficient uptake and degradation of Stx by macrophages and, consequently, for potent Stx-neutralizing activity in the circulation. We also determined the binding site of the SUPER TWIGs on Stx. PMID- 15897997 TI - Amelioration of the effects of Citrobacter rodentium infection in mice by pretreatment with probiotics. AB - BACKGROUND: Citrobacter rodentium is a naturally occurring murine pathogen that causes colonic epithelial-cell hyperplasia, disrupts the colonic mucosa, and elicits a predominantly T helper 1 cellular immune response; it thereby serves as a model for the study of mechanisms of disease induced by human attaching effacing pathogens. We sought to determine whether pretreatment of mice with a mixture of Lactobacillus rhamnosus and L. acidophilus probiotics would attenuate C. rodentium-induced colonic disease in mice. METHODS: Mice were administered sterile drinking water, probiotics (10(9) cfu/mL) in sterile drinking water, maltodextrin in sterile drinking water, orogastric C. rodentium (10(7) cfu in 0.1 mL), or maltodextrin in sterile drinking water for 1 week before C. rodentium infection, or they were pretreated with probiotics (10(9) cfu/mL) for 1 week before challenge with C. rodentium. RESULTS: Mice that received viable probiotics remained healthy. C. rodentium infection elicited mucosal inflammation, epithelial-cell hyperplasia, apoptosis in the colon, and interferon (IFN)- gamma production by splenocytes. Pretreatment with probiotics decreased levels of all but IFN- gamma production. CONCLUSIONS: Pretreatment with probiotics attenuates the effects of C. rodentium infection in mice. Understanding the mechanism of these beneficial effects will aid in determining the efficacy of probiotics in preventing infection with related attaching-effacing enteric pathogens in humans. PMID- 15897998 TI - Eagle effect in Corynebacterium diphtheriae. AB - The in vivo relevance of the paradoxical bactericidal effect (the Eagle effect) is not evident. We found in vitro a paradoxical bactericidal effect of amoxicillin on 2 strains of nontoxigenic Corynebacterium diphtheriae. Then, using an experimental rabbit model of endocarditis, we evaluated the in vivo relevance of this phenomenon. Rabbits were assigned to the following groups: no treatment (control group), continuous amoxicillin infusion simulating a dosage of 200 mg/kg/day in humans, and continuous amoxicillin infusion simulating a dosage of 20 mg/kg/day in humans. The low dosage (20 mg/kg/day) was significantly more effective than the high dosage (200 mg/kg/day) against both strains (P<.025), confirming the paradoxical bactericidal effect observed in vitro. PMID- 15897999 TI - Modulation of expression of superantigens by human transferrin and lactoferrin: a novel mechanism in host-Streptococcus interactions. AB - The role played by host-pathogen interactions in regulation of expression of streptococcal virulence factors in vivo is beginning to become clear. We have reported that the expression of 2 streptococcal virulence factors, the streptococcal pyrogenic exotoxin (Spe) A and the cysteine protease SpeB, was reciprocally modulated during infection with Streptococcus pyogenes. To identify host signals mediating this reciprocal regulation, we cocultured clonal M1T1 isolates with human peripheral blood mononuclear cells (PBMCs). In accordance with our in vivo findings, when bacteria were in direct contact with human PBMCs or were separated in transwells, expression of speA was induced, whereas expression of speB was down-regulated. This phenomenon was mediated by transferrin and lactoferrin and was influenced by the iron-saturation status of these proteins. Iron chelation from media induced expression of speA, but to a much lesser degree than did that with apotransferrin and lactoferrin, suggesting additional effects of these ferrins on modulation of expression of speA and speB. Thus, ferrins may play an important role in host-pathogen interactions in skin and mucosal tissues. PMID- 15898000 TI - A heterologous prime-boost vaccination regimen using ORFF DNA and recombinant ORFF protein confers protective immunity against experimental visceral leishmaniasis. AB - OBJECTIVE: We describe the effectiveness of a prime-boost vaccination regimen using the open-reading frame (ORFF) gene from the LD1 locus of Leishmania donovani. METHODS: A group of BALB/c mice was immunized with the plasmid carrying the gene for ORFF (F/pcDNA 3.1) and given a booster dose of either the same DNA vaccine or a vaccine with a recombinant ORFF (rORFF) protein. Another group of BALB/c mice was immunized and given a booster dose of the rORFF protein vaccine. The protective efficacies of these vaccine formulations were compared after challenge with L. donovani stationary-phase promastigotes. RESULTS: Mice given the prime-boost vaccination regimen had an enhanced reduction in parasite load (75%-80%), compared with that in mice given only the rORFF protein vaccine (45% 60%). However, the protective response induced in the prime-boost group was not more than that elicited in the DNA vaccine group. Immunization with only the rORFF protein vaccine did not induce the typical T helper response, whereas priming with the DNA vaccine resulted in enhanced production of immunoglobulin G2a and interferon- gamma . Furthermore, priming with the DNA vaccine also led to enhanced proliferation of splenocytes, suggesting subsequent expansion of antigen specific T cells. CONCLUSIONS: The heterologous prime-boost vaccination strategy may be utilized for visceral leishmaniasis. PMID- 15898001 TI - The cholinergic anti-inflammatory pathway regulates the host response during septic peritonitis. AB - BACKGROUND: The nervous system, through the vagus nerve, can down-regulate inflammation in vivo by decreasing the release of tumor necrosis factor- alpha by endotoxin-stimulated macrophages. This anti-inflammatory effect is mediated by an interaction between acetylcholine, the principal neurotransmitter of the vagus nerve, and cholinergic nicotinic acetylcholine receptors on macrophages. METHODS: We determined the role of this "cholinergic anti-inflammatory pathway" during septic peritonitis induced in mice by intraperitoneal injection of live Escherichia coli. Septic peritonitis was preceded by inhibition of the cholinergic anti-inflammatory pathway by unilateral cervical vagotomy, by stimulation of this pathway by pretreatment of mice with nicotine, or by a combination of both interventions. RESULTS: Initial cytokine release during septic peritonitis was enhanced after previous vagotomy and was decreased after nicotine pretreatment, independently of the integrity of the vagus nerve. Further study established that vagotomy before septic peritonitis resulted in an enhanced influx of neutrophils and a marked increase in proinflammatory cytokine levels and liver damage. Conversely, nicotine pretreatment strongly decreased cell influx, proinflammatory cytokine levels, and liver damage, whereas bacterial clearance and survival were impaired. DISCUSSION: These data provide the first evidence, to our knowledge, of an important role of the vagus nerve in regulating the innate immune response to a severe bacterial infection. PMID- 15898002 TI - Inhibition of daptomycin by pulmonary surfactant: in vitro modeling and clinical impact. AB - The lipopeptide daptomycin has been approved for use in skin and skin-structure infections but has failed to meet statistical noninferiority criteria in a clinical trial for severe community-acquired pneumonia. Daptomycin exhibited an unusual pattern of activity in pulmonary animal models: efficacy in Staphylococcus aureus hematogenous pneumonia and inhalation anthrax but no activity against Streptococcus pneumoniae in simple bronchial-alveolar pneumonia. Daptomycin was shown to interact in vitro with pulmonary surfactant, resulting in inhibition of antibacterial activity. This effect was specific to daptomycin and consistent with its known mechanism of action. This represents the first example of organ-specific inhibition of an antibiotic. PMID- 15898004 TI - Cytomegalovirus load in bronchoalveolar lavage fluid: a clue to the diagnosis of cytomegalovirus pneumonia? PMID- 15898005 TI - Estimating female-to-male infectivity of HIV-1 in Kenya: potential threats to validity. PMID- 15898007 TI - Persistent GB virus C infection is associated with decreased HIV-1 disease progression in the Amsterdam Cohort Study. PMID- 15898008 TI - GB virus C infection and survival in the Amsterdam Cohort Study. PMID- 15898010 TI - [Benign idiopathic external hydrocephalus (benign subdural collection) in 39 children: its natural history and relation to familial macrocephaly]. AB - INTRODUCTION: Benign idiopathic external hydrocephalus (BIEH) is an age-dependent disorder that is self-limiting in time and has an uncertain aetiology. PATIENTS AND METHODS: A retrospective study was conducted involving 39 patients (16 girls and 23 boys) with BIEH. The following data were analysed for each patient: age, sex, family history, history concerning pregnancy, childbirth and neonatal period, postnatal history, data from clinical records and from physical examinations, progress of psychomotor development, findings from the first and successive neuroimaging studies, results of other complementary examinations, clinical and neuroimaging situation at the last check-up that was carried out, length of clinical control, age at which subdural effusion disappeared, and any other relevant associated facts. RESULTS: Age at diagnosis ranged from 1.33 and 25 months (mean: 8.4 months); in 38.46% of cases there was a history of macrocephalia in one of the progenitors; in four of them the presence of congenital macrocephalia was noted; in five, there was motor retardation and one of them displayed psychomotor retardation; in 15, there was an association with a slight dilatation of the lateral ventricles; the mean time of clinical control was 3.36 years; the process was seen to resolve in 14 cases; the minimum age for the disappearance of the subdural effusion was 9 months and the maximum was 8 years; macrocephalia persisted until the clinical control ended in 22 of the cases. We also noted the presence of two cases of mitochondrial encephalomyopathy, one craniosynostosis of the sagittal suture, one microdeletion 22q11.2, one a-1 antitrypsin deficiency, and one case of idiopathic bilateral congenital palpebral ptosis. CONCLUSIONS: The subdural effusion and/or macrocephalia persist in a high percentage of these patients and sometimes there is a close relationship between this condition and benign familial macrocephalia. Despite its benignity, it can influence psychomotor or motor retardation and behavioural disorders. On rare occasions it may be associated to mitochondrial encephalomyopathy and to the microdeletion 22q11.2. PMID- 15898011 TI - [Evaluation of the survival of bone marrow mononucleate cells transplanted in a rat model of striatal lesion with quinolinic acid]. AB - INTRODUCTION: Transplant is one of the alternatives available for the treatment of neurodegenerative diseases aimed at replacing the cells lost during the course of the disease. One promising source of cells for the development of transplants could be the mononucleate cells from bone marrow. AIMS. The purpose of this study was to study the capacity of bone marrow mononucleate cells to survive the transplant process, and to search for a method that enables tracking of these cells in vivo once they have been implanted. MATERIALS AND METHODS: Bone marrow mononucleate cells were extracted from the femur of rats by means of a Ficoll Hypaque gradient. The cells under study were modified genetically with an adenovirus that expresses the PFV or which are marked with Hoechst dye. The marked cells were implanted in the striatum of rats with lesions caused by quinolinic acid. RESULTS: The viability of the genetically modified cells was low, whereas that of the cells marked with Hoechst dye was above 90%. The implanted cells survived the transplant at least a month and dispersed away from the site of entry towards the corpus callosum and cortex. CONCLUSIONS: We consider that the use of Hoechst dye offers more advantages for tracking these cells in vivo. Mononucleate cells have a number of characteristics that allow them to be included as candidate sources of cells for the treatment of neurodegenerative diseases. PMID- 15898012 TI - [Cognitive evoked potentials in uncomplicated hypertensive patients]. AB - INTRODUCTION: One of the most frequent complications of vascular encephalopathies is the onset of cognitive disorders that lead the patient to a progressive decay of his abilities and useful life. Being high blood pressure (HBP) one of the most important causes of brain vascular disorders, we studied some aspects of cognition in HBP patients without neurological manifestations and with normal scores in Mini Mental State Examination. PATIENTS AND METHODS: There were studied 84 uncomplicated hypertensive patients, with a normal neurological physical examination and a group of 35 functionally healthy subjects. The patients were divided into three groups: slight high blood pressure (SLHBP, n = 24 age, 49 +/- 4) with diastolic blood pressure (DBP) between 90 and 100 mmHg, moderate high blood pressure (MHBP, n = 40, age 51 +/- 6) with DBP between 101 and 114 mmHg, and severe high blood pressure (SHBP, n = 20, age 49 +/- 8) with DBP of 115 mmHg or higher. All subjects underwent visual and auditory P300 evoked potentials. RESULTS: Visual and auditory P300 wave showed global latency delay, and amplitude decrease and absence of response in different leads, especially in frontal regions, higher in SHBP patients. This absence of response was more evident in auditory P300 in all patient groups. CONCLUSION: It is remarkable that neurologically asymptomatic HBP patients have P300 alterations with regional focalization, possibly due to a functional disorder related to hemodynamic changes in the central nervous system. PMID- 15898013 TI - [Delays before and after arrival at the hospital in the treatment of strokes]. AB - INTRODUCTION: Only a minority of patients with acute stroke are eligible for fibrinolysis, and the main reason for this is that time runs out and goes beyond the therapeutic window. The chief delay occurs prior to arrival at the hospital, but there is also a nosocomial delay, which has received far less attention. AIMS: The purpose of our study was to describe the data on delays occurring before and after arrival at the hospital in a sample of patients with acute stroke, and to analyse possible associated factors, including the overload on health care services in the Emergency department, an aspect that has not previously been evaluated. PATIENTS AND METHODS: Epidemiological data, times of delays before and after entering the hospital, means of transport used and the health care workload in the Emergency department of the day of admission were collected for all the stroke patients admitted to the Stroke Unit of the Hospital General Universitario de Alicante throughout the period under study. RESULTS: From an initial sample of 460 patients, 423 were finally included in the study, with a mean delay before and after admission to hospital of 3.99 and 2.36 hours, respectively. Use of an ambulance and the haemorrhagic and transient ischemic attack subtypes were linked to a shorter delay before reaching the hospital, and arrival during the night shift was the only factor associated to a longer delay once inside the hospital. CONCLUSIONS: In our environment, there is a need to establish strategies aimed at shortening the times stroke patients take to reach the hospital and the time that elapses before they are attended. PMID- 15898014 TI - [Herpes simplex encephalitis in Caceres. An analysis of the last five years]. AB - INTRODUCTION: Herpes simplex encephalitis (HSE) is a rare disease, although it is still the most common serious viral infection of the central nervous system. Little is currently known about its epidemiology and clinical features in our area. This paper presents a retrospective description of the clinical, diagnostic and progressive characteristics of patients diagnosed with HSE in Caceres General Hospital, and confirmed by means of a polymerase chain reaction (PCR) study for the DNA of the herpes simplex virus over the last 5 years. CASE REPORTS: We found four patients who had been diagnosed with HSE in the last 5 years (3.3 cases/million inhabitants/year; CI at 95% of 2.42-4.18), two males and two females, with an age interval between 58 and 75 years. All the patients had high temperatures and three of them also presented neurological focus. A computerised axial tomography scan of the head performed on admission was interpreted as being normal in all cases, while magnetic resonance imaging of the head carried out in two patients showed lesions compatible with HSE in both cases. Electroencephalographic studies were carried out in two patients and revealed focusing in one of them and severe generalised involvement in the other. Analysis of the cerebrospinal fluid (CSF) disclosed lymphocytic meningitis in four cases, although in one of them the study was normal on admission. In two of the patients, viral meningoencephalitis was suspected from the moment the patient was admitted to hospital. For this reason, early treatment with IV acyclovir was established and the clinical progression was good in both patients, although one of them was left with mild neurological sequelae. The other two cases, who died, had a severe underlying pathology and alternative clinical diagnoses that were reasonable on admission (extensive pneumonia and delirium tremens in one of them, and hypercapnic encephalopathy with severe chronic obstructive pulmonary disease in the other), and the clinical suspicion of HSE and the establishment of treatment were late. CONCLUSIONS: The incidence of HSE in our environment is in the upper interval of that reported in the literature. PCR analysis for the herpes simplex virus in CSF seems to have replaced brain biopsy studies as the diagnostic technique. The underlying severe chronic pathology makes it more difficult to diagnose and worsens the prognosis. A multicentre epidemiological study should be conducted to confirm these findings. PMID- 15898015 TI - [Horner syndrome as the first symptom of nasopharyngeal cancer. Two case reports]. AB - INTRODUCTION: Horner syndrome (HS) is produced by damage to the oculosympathetic pathway and gives rise to palpebral ptosis, enophthalmos and myosis. Anhidrosis also occurs in cases in which the lesion is located before the separation of the vasomotor and sudomotor fibres at the start of the internal carotid artery. Nasopharyngeal cancer may damage the cranial nerves, mainly in the cavernous sinus, and very few cases of HS due to infiltration of the tumour into the parapharyngeal space have been reported. CASE REPORTS: Case 1: a 67-year-old male with a 10-day history of HS on the left side without anhidrosis. The aetiological study revealed a nasopharyngeal lymphoepithelioma; the tumour became smaller and HS disappeared following treatment with chemotherapy. Case 2: a 48-year-old male with a 1-week history of HS without anhidrosis, and a 2-month history of deafness and pain in the right ear. The aetiological study showed a nasopharyngeal lymphoepithelioma and serous otitis. The tumour became smaller and HS disappeared following treatment with chemotherapy. CONCLUSIONS: It is important to carry out studies of the nasopharynx in patients with this syndrome to allow early diagnosis and treatment of a region that is not readily available for direct examination. The existence of an associated pathology in the ear or the paranasal sinuses strongly suggests that its origins lie in the nasopharynx. PMID- 15898016 TI - [Treatment of racemose neurocysticercosis of the spine]. AB - INTRODUCTION: Neurocysticercosis (NCC) is the most frequent parasitic infection of the central nervous system, and its prevalence is high in Portugal. Spinal involvement is rare, only occurring in between 1 and 5% of cases, and causes many problems when it comes to its diagnosis and treatment. Spinal leptomeningeal NCC is particularly difficult to treat, especially if associated to arachnoiditis. We report a case of NCC with spinal leptomeningeal involvement associated to extensive arachnoiditis, in which the therapeutic measures we have available today were totally ineffective. CASE REPORT: We describe the case of a 37-year old male from Cape Verde, who was kept under surveillance because of a 3-month history of symptoms of cauda equina syndrome. In the last few months he also complained of holocranial headaches. Magnetic resonance (MR) imaging showed numerous cystic lesions in the bottom of the thecal sac, associated to extensive arachnoiditis, and MR images of the brain revealed compensated hydrocephalus. Diagnosis of NCC was confirmed by positive immunoblot in serum and in cerebrospinal fluid (CSF). Following insertion of a ventriculoperitoneal shunt, the patient received treatment with albendazole and corticoids for two weeks, and showed a clinical improvement. One year later, he showed symptoms of acute tetraparesis, and a cervical MRI showed cystic lesions with spinal cord compression and intense arachnoiditis. In spite of a decompression laminectomy, with removal of the cysts, and therapy with praziquantel and corticoids, the patient's clinical status did not improve. CONCLUSIONS: Spinal racemose NCC associated to arachnoiditis has a poor prognosis and is often untreatable. The therapeutic options in NCC of the spine are discussed, and attention is drawn to the importance of cysticercotic arachnoiditis in the prognosis of the disease. PMID- 15898017 TI - [Interrelations between sleep and the immune status]. AB - INTRODUCTION: The idea of the healing effects of the sleep over the disease is quite extended. Besides, the sleep and the circadian rhythms cause deep changes on the immune function. Reciprocally, the sleep also suffers deep changes when the immune system is challenged during an external aggression. DEVELOPMENT: This review shows some of the data supporting both observations. From the relationships between the sleep and the immune system, it has been proposed that one function of sleep is just to support the immune defense. However, an important fraction of the relationships between sleep and immune function might be a response to the stress produced both during the sleep disorders and when the organism activates the immune defense. Moreover, the epidemiological evidence only shows negligible results when contrasting the amount of sleep and the life expectancy. CONCLUSION: It seems thus probable that the relationships between sleep and immune function are only a reflect of additional factors, such as stress, which cause deep changes in sleep and immunity. PMID- 15898018 TI - [Visuoperceptual processing in Parkinson's disease: from the retina to the frontal cortex]. AB - INTRODUCTION: The growing interest in the cognitive impairment shown by patients with Parkinson's disease has led to a wealth of research in this line over recent years. In this paper we review the visuospatial alterations in these patients, which are usually linked to other disorders such as those affecting planning, sequencing, attention or mnemonic processes. DEVELOPMENT: We report the most relevant findings, which suggest that the existence of these visuospatial disorders shown by patients with Parkinson's disease are not always secondary to other frontal-type cognitive impairments, as has been claimed in recent years. Instead, they may be due to disorders in other anterior points of the perceptive process (as a result of dopaminergic deficits in basal-thalamic-cortical circuits). Thus, visuoperceptual disorders are classified according to their location in the brain: from the retina to the lateral geniculate nucleus, the visual cortex and the extrastriate cortex and, finally, the frontal and prefrontal cortex. CONCLUSIONS: We propose this classification of the disorders according to their location to aid in achieving an objective selection of the sample and of the neuropsychological tests used in studies. In this regard, we consider that there should be a higher degree of agreement among researchers when it comes to designing research projects that deal with visuospatial disorders in patients with Parkinson's disease. PMID- 15898019 TI - [The Andalusia Epilepsy Society's Guide to Epilepsy Therapy 2005: I. Criteria for starting and discontinuing and the pharmacological bases of antiepileptic treatment]. AB - AIMS: The objective of this work was to produce a scientific evidence-based guide to clinical practice dealing with the basic questions concerning the treatment of epilepsy. DEVELOPMENT: A committee of 11 experts belonging to the Andalusia Epilepsy Society, made up of six neurologists, three neuropaediatricians, one neurosurgeon and a pharmacologist, all of whom were deeply involved and experienced in epilepsy, conducted a thorough review of the literature in search of all the evidence available on the proposed subject matter. The following databases were used: MEDLINE, Cochrane Library and the databases of several clinical practice guidelines (National Guideline Clearinghouse, National Institute of Clinical Excellence and the American Academy of Neurology's Clinical Guidelines). The Guide was set out in seven sections and was published in four parts. From a total number of 187 relevant documents, the committee found 63 examples of scientific evidence and 91 therapeutic recommendations. These were tabulated and classified according to the European Federation of Neurological Societies' criteria for producing Clinical Practice Guidelines. CONCLUSIONS: The results of this survey provide scientific evidence-based clinical guidelines that are useful, simple and applicable at different levels of health care. PMID- 15898020 TI - [Early diagnosis of primary progressive aphasia by positron emission tomography]. PMID- 15898021 TI - [Accidental methyl alcohol poisoning: acute and chronic neurological sequelae]. PMID- 15898022 TI - Fellowship training in regional anesthesia. PMID- 15898023 TI - Guidelines for regional anesthesia fellowship training. AB - BACKGROUND: The number of regional anesthesia fellowships has grown over the past 2 decades. There currently exist no guidelines for what constitutes ideal regional anesthesia fellowship training. METHODS: Regional anesthesia fellowship program directors and other advocates of regional anesthesia were invited to participate in a collaborative project to establish a standardized curriculum for regional anesthesia fellowships. Guidelines were created based on the existing template of Accreditation Council of Graduate Medical Education program requirements for residency education in anesthesiology. The resulting draft guidelines were distributed at a meeting of the program directors, who were then asked to forward all comments and relevant training material from their respective institutions to a coordinating institution. RESULTS: All received materials were reviewed, and selected components were collated into a consensus document, which was then reviewed, modified, and eventually approved by the program directors over a 2-year series of meetings. The program directors agreed to adopt the guidelines as their fellowship curriculum and to evaluate their effectiveness in 2 years' time. CONCLUSIONS: The intent of these initial guidelines is to improve the quality and consistency of regional anesthesia fellowship training. The creation process also led to an affirmation of the directors' commitment to continued dialogue for the purpose of facilitating the exchange of ideas among programs. PMID- 15898024 TI - The training and careers of regional anesthesia fellows--1983-2002. AB - BACKGROUND AND OBJECTIVES: The education and subsequent careers of regional anesthesia fellows have not been examined but may provide insight into improving future fellowship training and/or the future of the subspecialty. METHODS: Regional anesthesia fellows educated during a 20-year period (1983-2002) were asked to complete a comprehensive survey that detailed their training, current professional setting, and use of regional anesthesia, and how they foresee the future of regional anesthesia. A separate survey of academic anesthesiology chairs assessed the role of and need for regional anesthesiologists in teaching departments. RESULTS: Twelve regional anesthesia fellowship programs in the United States and Canada provided contact information on 176 former fellows. The survey response rate from those practicing in North America was 49% (77/156). Two of the 12 responding institutions have trained 68% of regional anesthesia fellows. Of respondents, 61% are or have been in academic practice. Regional anesthesia remains an integral part of most respondents' current practice, as evidenced by significant use of regional techniques, active involvement in subspecialty societies, and participation in continuing medical education programs. Academic chairs indicate that fellowship-trained regional anesthesiologists play important roles in resident education and are in demand by academic departments. CONCLUSIONS: This report details how regional anesthesia fellows from 1983 to 2002 were trained and how they currently practice and examines their insights regarding the strengths and weaknesses of past and future regional anesthesia education. PMID- 15898025 TI - Resident versus staff anesthesiologist performance: coracoid approach to infraclavicular brachial plexus blocks using a double-stimulation technique. AB - OBJECTIVES: Infraclavicular brachial plexus block with double stimulation (ICB) is a safe technique for upper-limb anesthesia. However, the experience of learning this technique by anesthesiology residents has not been reported. The aim of this study was to compare staff with resident anesthesiologists in the performance of ICB. METHODS: Patients scheduled for orthopedic surgery of the upper limb were included in a prospective, comparative, randomized study and were given ICB by either staff anesthesiologist (Group S, n = 110 patients) or resident anesthesiologist (Group R, n = 110 patients). RESULTS: Time to perform the block was 3.9 minutes (95% confidence interval [CI 95%] = 3.5 to 4.3) for Group S and 5.8 minutes (CI 95% = 5.2 to 6.4) for Group R (P < .05). The onset time was 14.4 minutes (CI 95% = 13.5 to 15.3) for Group S and 15.9 minutes (CI 95% = 14.7 to 17.1) for Group R (P = NS). Success rate was 93% for Group S and 90% for Group R (P = NS). Supplementation was performed in 8 patients in Group S versus 11 patients in Group R (P = NS). No patient needed general anesthesia. One self-limited vascular puncture was made in Group S versus 3 in Group R (P = NS). CONCLUSION: This report determines whether residents can perform this technique with comparable efficiency compared with staff. We conclude that ICB should be taught as part of all resident training programs. PMID- 15898026 TI - Clinical efficacy of the brachial plexus block via the posterior approach. AB - BACKGROUND AND OBJECTIVES: The posterior approach to the brachial plexus remains underused. We assessed the clinical effectiveness of this technique for shoulder surgery. METHODS: One hundred eighty-seven consecutive patients scheduled to undergo shoulder surgery were assessed after a single-injection nerve-stimulation technique using ropivacaine 0.75%. Sensory and motor block was evaluated in the distribution area of each terminal branch of the brachial plexus every 10 minutes for 30 minutes. Postoperative analgesia was evaluated at regular time intervals at rest and with passive movement, up to 24 hours postoperatively. RESULTS: The brachial plexus was reached at a depth of 6.5 +/- 0.9 cm. One attempt was sufficient in 85% of patients. Neck pain during insertion of the needle was encountered in 6 (3%; 95% confidence interval [CI], 0.7%-5.6%) patients. Thirty minutes after ropivacaine injection, the axillary, radial, median, musculocutaneous, and ulnar nerves were anesthetized in 100%, 100%, 97%, 96%, and 68% of cases, respectively. The success rate of the block was 98%. Postoperative analgesia was satisfactory in 97% of patients up to 12 hours after the initial injection. Dysphonia and Horner's syndrome were observed in 14 (7%; 95% CI, 3.7% 11.2%) and 12 (6%; 95% CI, 2.9%-9.9%) patients, respectively. One patient (0.5%; 95% CI, 0%-1.5%) had documented hemidiaphragmatic paresis. No complication was noted during the 3-month follow-up period. CONCLUSIONS: This study reports the clinical effectiveness of the single-injection nerve-stimulation technique for the brachial plexus block via the posterior approach in patients undergoing shoulder surgery. It appears to be effective, relatively safe, and well tolerated. PMID- 15898027 TI - Peripheral block of the hyperpolarization-activated cation current (Ih) reduces mechanical allodynia in animal models of postoperative and neuropathic pain. AB - BACKGROUND AND OBJECTIVES: Block of the hyperpolarization-activated inward current (I h) reduces excitability of peripheral axons during stimulation and decreases ectopic discharges in axotomized sensory neurons. Changes in I h expression in DRG neurons have been suggested to partially underlie sensitization after nerve injury and inflammation. We hypothesized that peripheral block of I h on axons would produce an antiallodynic effect in postoperative as well as neuropathic conditions, and we tested perineural administration of ZD 7288, a specific blocker of I h , on pain-associated behavior in animal models of neuropathic and postoperative pain. METHODS: Under halothane anesthesia, partial sciatic nerve injury or hind-paw incision were performed on adult male rats as previously described. Mechanical allodynia was inferred by demonstration of a decrease in paw withdrawal threshold by application of calibrated von Frey filaments. After surgery, animals received either a saline or a ZD 7288 solution either by sciatic perineural injection or by intraplantar injection. RESULTS: Perineural administration of ZD 7288 (100 microM) significantly reduced mechanical allodynia induced by partial sciatic nerve injury and hind-paw incision. Saline and 10 microM of ZD 7288 had no significant effect on mechanical allodynia. Contralateral administration of ZD 7288, 100 microM, did not affect ipsilateral paw withdrawal threshold after nerve injury. Intraplantar injection of ZD 7288 failed to reduce mechanical allodynia after nerve injury. Sedation and motor effects were not observed. CONCLUSIONS: The current study shows that peripheral block of I h produces an antiallodynic effect, which suggests that I h channels represent a novel target for nerve block treatment of postoperative and neuropathic pain. PMID- 15898028 TI - Intrathecal sufentanil is more potent than intravenous for postoperative analgesia after total-hip replacement. AB - BACKGROUND AND OBJECTIVES: In our clinical experience, sufentanil is more effective when administered intrathecally than intravenously. To test this hypothesis, we compared the analgesic characteristics of 7.5 microg of intrathecal or intravenous sufentanil for pain relief after total-hip replacement. METHODS: A randomized, double-blind study was conducted of 40 patients older than 75 years who experienced total-hip arthroplasty in which continuous spinal anesthesia was administered. In the recovery room, as soon as a pain score higher than 3 on a scale of 10 on a visual analog scale was reported, either 7.5 microg intrathecal or 7.5 microg intravenous sufentanil were given. If the pain score remained higher than 3 at 20 minutes after sufentanil administration,1.25 mg of "rescue" intrathecal bupivacaine were given. RESULTS: During the first 20 minutes after intrathecal or intravenous injection, a significantly faster relief of pain was observed for the intrathecal group from 2.5 until 20 minutes. Significantly, more patients needed rescue bupivacaine in the intravenous group (7 of 20 v 0 of 20, P < .008), whereas significantly more patients in the intrathecal group reached a pain score of 0 (20 of 20 v 9 of 20, P < .001). The time to the first analgesic intervention for a pain score greater than 3 was significantly longer in the intrathecal group (224 +/- 100 v 98 +/- 60 minutes, P < .001). Pruritus was observed only in 5 patients of the intrathecal group (P < .047), whereas peripheral oxygen saturation under 95% was observed only in 6 patients in the intravenous group (P < .045). CONCLUSIONS: After total hip replacement, intrathecal route of sufentanil administration rapidly offers excellent analgesia of better quality and longer duration when compared with the intravenous route. PMID- 15898029 TI - Medial canthus single-injection peribulbar anesthesia: a prospective randomized comparison with classic double-injection peribulbar anesthesia. AB - BACKGROUND AND OBJECTIVES: The authors report the first prospective randomized comparison of the medial canthus single-injection peribulbar anesthesia (also called caruncular anesthesia) with the classic double-injection peribulbar technique. METHODS: One hundred patients scheduled for cataract surgery were randomly assigned to either a single medial canthus injection or a double peribulbar injection of mepivacaine 2%. The amount of anesthetic agent injected was clinically adapted to each patient. Akinesia, volume injected, pain, reinjections, and complications were assessed after the procedure. RESULTS: The medial canthus single-injection peribulbar anesthesia was significantly less painful and required less anesthetic agent than the double-injection peribulbar anesthesia. Akinesia score and the reinjection rate were similar in the 2 groups, whereas chemosis was significantly more frequent in the double-injection group. CONCLUSIONS: Medial canthus single-injection peribulbar anesthesia appears to be an effective alternative to the usual double-injection peribulbar anesthesia. PMID- 15898030 TI - Two-pore domain potassium channels: new sites of local anesthetic action and toxicity. AB - Potassium (K+) channels form the largest family of ion channels with more than 70 such genes identified in the human genome. They are organized in 3 superfamilies according to their predicted membrane topology: (1) subunits with 6 membrane spanning segments and 1-pore domain, (2) subunits with 2 membrane-spanning segments and 1-pore domain, and (3) subunits with 4 membrane-spanning segments and 2-pore domains arrayed in a tandem position. The last family has most recently been identified and comprises the so-called 2-pore domain potassium (K2P) channels, believed responsible for background or leak K+ currents. Despite their recent discovery, interest in them is growing rapidly with more than 270 references in the literature reported (www.ipmc.cnrs.fr/~duprat/2p/ref2p.htm#2P, accessed October 30, 2004). K2P channels are widely expressed in the central nervous system and are involved in the control of the resting membrane potential and the firing pattern of excitable cells. This article will therefore review recent findings on actions of local anesthetics with respect to 2P channels. It begins with an overview of the role of background K+ channels in neuronal excitability and nerve conduction and is followed by a description of the K2P channel family including experimental evidence for the contribution of K2P channels to the mechanism of action and toxicity of local anesthetics. PMID- 15898031 TI - Extended-duration analgesia: update on microspheres and liposomes. PMID- 15898032 TI - The impact of technology on the analgesic gap and quality of acute pain management. AB - National surveys continue to document the undertreatment of acute postoperative pain, despite the availability of evidence-based, clinical practice guidelines and the Joint Commission on Accreditation of Healthcare Organizations standards. This article surveys factors that contribute to persistent gaps during the acute pain management process, including deficiencies in providing continuous analgesia, disparities in access to medical care, the acute pain medicine culture itself, a lack of adequate pain assessment, health care provider biases, and limited health care resources. The role of technology in increasing patients' control over their own pain management and narrowing these gaps is discussed. Patient-controlled analgesia delivery systems are an example of such technology, and they play a key role in improving the quality of acute pain management and increasing the patient's involvement in this process. However, the use of these systems may be limited because of the amount of health care resources necessary for their administration and limitations in payment for professional services. Therefore, there exists a need for additional technologies that will simplify the pain management process and reduce the amount of health care resources necessary to provide patients with quality acute pain management. PMID- 15898033 TI - Iontophoresis: the process behind noninvasive drug delivery. PMID- 15898034 TI - An imaging review of sacroiliac joint injection under computed tomography guidance. PMID- 15898035 TI - Rapid onset of massive subdural anesthesia. AB - OBJECTIVE: This case report describes the accidental placement of a lumbar labor epidural catheter into the subdural space resulting in the rapid onset of massive subdural anesthesia. CASE REPORT: A single-orifice open-ended catheter was accidentally placed in the lumbar subdural space. After a test dose of 3 mL 1.5% lidocaine with 15 microg of epinephrine and 2 minutes after a bolus of 5 mL of 0.25% bupivacaine, there was a rapid onset of massive subdural anesthesia with a loss of consciousness, respiratory arrest, and absent pulses. The subdural catheter placement was confirmed by injection of a contrast agent under fluoroscopy. CONCLUSION: Massive subdural anesthesia is a complication of epidural catheter placement and may have a rapid onset causing a loss of consciousness, respiratory arrest, and absent pulses. PMID- 15898036 TI - A modified approach to transcrural celiac plexus block. AB - OBJECTIVES: Transcrural celiac block using the needle "walking off" the L1 vertebra technique may cause complications. We used patient-specific computed tomography (CT) images as a roadmap to perform the block under fluoroscopy. We present 1 case to describe the technique. CASE REPORT: The patient is a 63-year old woman with refractory pain from pancreatic cancer. Her CT showed the celiac trunk at the upper L1 vertebra and 2 cm left to the midline. Needle trajectories were drawn on that film. The line representing the classic "walking off" the bone technique on the left side crossed the aorta. Two lines targeting the base of the celiac trunk were modified, thereby avoiding both the L1 vertebra and the surrounding organs. The following were measured: the distance from the midline to the left needle entry (2.5 cm), the angle for the left needle insertion (90 degrees), the distance (6 cm) and the angle (65 degrees) for the right needle entry, and the distance from the anterior margin of the L1 to the celiac trunk (2.6 cm). During the procedure, 2 needles were placed according to these measurements in a plane superior to the transverse process of the L1. No bony contact or needle redirection was made. Both needles reached 3 cm anterior to the anterior margin of the L1. X-ray contrast crossed the midline and silhouetted the target vasculature. Five milliliters of 0.2% ropivacaine followed by 10 mL of 6% phenol were injected on each side. The patient's pain level improved to 0 to 1/10 on a visual analog scale. CONCLUSIONS: The modified technique avoided painful needle contact on the bone, reduced needle redirections, and decreased the possibility of vital organ puncture. PMID- 15898037 TI - Re: combination of intraneural injection and high-injection pressure leads to fasicular injury and neurologic deficits in dogs. PMID- 15898038 TI - Unilateral tremor of the upper and lower limb after an axillary brachial plexus block. PMID- 15898040 TI - Use of colored syringes reduces the incidence of syringe swap during anesthesia. PMID- 15898041 TI - Veterans Administration-National Surgical Quality Improvement Program. PMID- 15898042 TI - Labeling as a tool to reduce drug error in anesthesia practice. PMID- 15898044 TI - High potential for epidural analgesia neuraxial block-associated hypotension in conjunction with heated intraoperative intraperitoneal chemotherapy. PMID- 15898045 TI - Maximum recommended doses of local anesthetics: a constant cause of confusion. PMID- 15898046 TI - A similar case of traumatic syrinx after implantation of an intrathecal catheter. PMID- 15898047 TI - "Postdural" is an ambiguity. PMID- 15898056 TI - Trannulenes with "in-plane" aromaticity: candidates for harvesting light energy. PMID- 15898053 TI - Galphas protein C-terminal alpha-helix at the interface: does the plasma membrane play a critical role in the Galphas protein functionality? AB - The heterotrimeric guanine nucleotide-binding regulatory proteins (G proteins, Galphabetagamma) mediate the signalling process of a large number of receptors, known as G protein-coupled receptors. The C-terminal domain of the heterotrimeric G protein alpha-subunit plays a key role in the selective activation of G proteins by their cognate receptors. The interaction of this domain can take place at the end of a cascade including several successive conformational modifications. Galpha(s)(350-394) is the 45-mer peptide corresponding to the C terminal region of the Galpha(s) subunit. In the crystal structure of the Galpha(s) subunit it encompasses the alpha4/beta6 loop, the beta6 beta-sheet segment and the alpha5 helix region. Following a previous study based on the synthesis, biological activity and conformational analysis of shorter peptides belonging to the same Galpha(s) region, Galpha(s)(350-394) was synthesized and investigated. The present study outlines the central role played by the residues involved in the alpha4/beta6 loop and beta6/alpha5 loops in the stabilization of the C-terminal Galpha(s)alpha-helix. H(2)O/(2)H(2)O exchange experiments, and NMR diffusion experiments show interesting evidence concerning the interaction between the SDS micelles and the polypeptide. These data prompt intriguing speculations on the role of the intracellular environment/cellular membrane interface in the stabilization and functionality of the C-terminal Galpha(s) region. PMID- 15898057 TI - Epigenetics--an epicenter of gene regulation: histones and histone-modifying enzymes. AB - The treatment of cancer through the development of new therapies is one of the most important challenges of our time. The decoding of the human genome has yielded important insights into the molecular basis of physical disorders, and in most cases a connection between failures in specific genes and the resulting clinical symptoms can be made. The modulation of epigenetic mechanisms enables, by definition, the alteration of cellular phenotype without altering the genotype. The information content of a single gene can be crucial or harmful, but the prerequisite for a cellular effect is active gene transcription. To this end, epigenetic mechanisms play a very important role, and the transcription of a given gene is directly influenced by the modification pattern of the surrounding histone proteins as well as the methylation pattern of the DNA. These processes are effected by different enzymes which can be directly influenced through the development of specific modulators. Of course, all genetic information is written as a four-character code in DNA. However, epigenetics describes the art of reading between the lines. PMID- 15898058 TI - Nitro(nitroso)cyanomethanides. PMID- 15898059 TI - Selective electrochemical detection of hydrogen fluoride by ambiphilic ferrocene derivatives. PMID- 15898060 TI - ERbeta protein expression in female cynomolgus monkey and CF-1 mouse brain: Western analysis. AB - In humans and rodents, multiple ERbeta variants with sizes ranging from 477-549 amino acids (aa) have been described. The identification of these variants in target tissues has important implications for estrogen signaling and cellular responsiveness. Western blot analysis using two anti-ERbeta antibodies specific for mammalian ERbeta sequences (PA1-310B and PA1-311) was employed to examine ERbeta protein expression in neural tissues from ovariectomized (OVX) cynomolgus macaques and CF-1 mice as well as to assess potential regulatory effects of acute and extended estradiol (E(2)) treatment. In hypothalamic extracts from both species, a single ERbeta immunoreactive (ERbeta-ir) band was detected at approximately 54 kDa, corresponding to the expected molecular weight for ERbeta477 and/or 485. In cynomolgus females, oral E(2) administration for 16 weeks had no apparent effect on hypothalamic ERbeta protein expression. In mouse, a single injection of E(2) did not change hypothalamic ERbeta protein levels 1.5, 4, 8, 16, or 24 h after injection. Extending the hormonal treatment to 4 or 21 days in OVX female mice also had no effect on the level of hypothalamic ERbeta protein. Additional regional analyses in female mouse brain with PA1-310B antibody showed that a second, 59 kDa ERbeta-ir band was present in cortex, striatum, hippocampus, and amygdala that could represent one or both of the larger ERbeta variants (530 and 549aa). The expression level of the second ERbeta isoform exhibited regional variation, with the strongest immunoreactivity detected in cortex and amygdala. Elucidating the functions of these ERbeta isoforms in the CNS will facilitate our understanding of the tissue- and promoter specific actions of estrogen. PMID- 15898061 TI - Chorda tympani nerve transection at different developmental ages produces differential effects on taste bud volume and papillae morphology in the rat. AB - Chorda tympani nerve transection (CTX) results in morphological changes to fungiform papillae and associated taste buds. When transection occurs during neonatal development in the rat, the effects on fungiform taste bud and papillae structure are markedly more severe than observed following a comparable surgery in the adult rat. The present study examined the potential "sensitive period" for morphological modifications to tongue epithelium following CTX. Rats received unilateral transection at 65, 30, 25, 20, 15, 10, or 5 days of age. With each descending age at the time of transection, the effects on the structural integrity of fungiform papillae were more severe. Significant losses in total number of taste buds and filiform-like papillae were observed when transection occurred 5-30 days of age. Significant reduction in the number of taste pores was indicated at every age of transection. Another group of rats received chorda tympani transection at 10, 25, or 65 days of age to determine if the time course of taste bud degeneration differed depending on the age of the rat at the time of transection. Taste bud volumes differed significantly from intact sides of the tongue at 2, 8, and 50 days post-transection after CTX at 65 days of age. Volume measurements did not differ 2 days post-transection after CTX at 10 or 25 days of age, but were significantly reduced at the other time points. Findings demonstrate a transitional period throughout development wherein fungiform papillae are highly dependent upon the chorda tympani for maintenance of morphological integrity. PMID- 15898062 TI - Role of the neuropeptide CCAP in Drosophila cardiac function. AB - The heartbeat of adult Drosophila melanogaster displays two cardiac phases, the anterograde and retrograde beat, which occur in cyclic alternation. Previous work demonstrated that the abdominal heart becomes segmentally innervated during metamorphosis by peripheral neurons that express crustacean cardioactive peptide (CCAP). CCAP has a cardioacceleratory effect when it is applied in vitro. The role of CCAP in adult cardiac function was studied in intact adult flies using targeted cell ablation and RNA interference (RNAi). Optical detection of heart activity showed that targeted ablation of CCAP neurons selectively altered the anterograde beat, without apparently altering the cyclic cardiac reversal. Normal development of the abdominal heart and of the remainder of cardiac innervation in flies lacking CCAP neurons was confirmed by immunocytochemistry. Thus, in addition to its important role in ecdysis behavior (the behavior used by insects to shed the remains of the old cuticle at the end of the molt), CCAP may control the level of activity of the anterograde cardiac pacemaker in the adult fly. Expression of double stranded CCAP RNA in the CCAP neurons (targeted CCAP RNAi) caused a significant reduction in CCAP expression. However, this reduction was not sufficient to compromise CCAP's function in ecdysis behavior and heartbeat regulation. PMID- 15898063 TI - Neuronal calcium sensor-1 gene ncs-1a is essential for semicircular canal formation in zebrafish inner ear. AB - We have analyzed the functional role of neuronal calcium sensor-1 (Ncs-1) in zebrafish development. We identified two orthologs of the mammalian NCS-1 gene. Full-length cDNAs encoding zebrafish Ncs-1a and Ncs-1b polypeptides were cloned and characterized. Whole-mount in situ hybridization revealed that ncs-1a mRNA was expressed beginning at early somitogenesis. As development progressed, ncs-1a mRNA was present throughout the embryo with expression detected in ventral hematopoietic mesoderm, pronephric tubules, CNS nuclei, and otic vesicle. By 4.5 days post fertilization (dpf), ncs-1a expression was detected primarily in the brain. Expression of ncs-1b mRNA was first detected at 36 hours post fertilization (hpf) and was restricted to the olfactory bulb. By 4.5 dpf, ncs-1b was expressed at low levels throughout the brain. Knockdown of ncs-1a mRNA translation with antisense morpholinos blocked formation of semicircular canals. These studies identify a novel function for ncs-1a in inner ear development and suggest that this calcium sensor plays an important role in vestibular function. PMID- 15898065 TI - Immediate early gene (ZENK, Arc) expression in the auditory forebrain of female canaries varies in response to male song quality. AB - In male songbirds, the song control pathway in the forebrain is responsible for song production and learning, and in females it is associated with the perception and discrimination of male song. However, experiments using the expression of immediate early genes (IEGs) reveal the activation of brain regions outside the song control system, in particular the caudomedial nidopallium (NCM) and the caudomedial mesopallium (CMM). In this study on female canaries, we investigate the role of these two regions in relation to playback of male songs of different quality. Male canaries produce elaborate songs and some contain syllables with a more complex structure (sexy syllables) that induce females to perform copulation solicitation displays (CSD) as an invitation to mate. Females were first exposed to playback of a range of songs of different quality, before they were finally tested with playback of songs containing either sexy or nonsexy syllables. We then sectioned the brains and used in situ hybridization to reveal brain regions that express the IEGs ZENK or Arc. In CMM, expression of ZENK mRNA was significantly higher in females that last heard sexy syllables compared to those that last heard nonsexy syllables, but this was not the case for NCM. Expression of Arc mRNA revealed no differences in either CMM or NCM in both experimental groups. These results provide evidence that in female canaries CMM is involved in female perception and discrimination of male song quality through a mechanism of memory reconsolidation. The results also have further implications for the evolution of complex songs by sexual selection and female choice. PMID- 15898066 TI - Effects of forest structure and composition on food availability for Varecia variegata at Ranomafana National Park, Madagascar. AB - We present a summary of a long-term field study that examined the effects of forest disturbance on the availability of palatable fruit and its utilization by V. variegata. Forest structure and tree species composition were measured in three adjacent study areas, with different histories of disturbance, in Ranomafana National Park (RNP), Madagascar. V. variegata abundance was monitored by frequent encounters with resident groups and periodic censuses conducted along trails. Finally, the abundance of mature fruit in species used by V. variegata was scored monthly at representative trees at several locations. V. variegata abundance was most consistent in the least anthropogenically disturbed site, while no established lemur groups were observed in the heavily logged site for over a decade post-harvest. Lemur abundance was variable in the selectively logged site. The presence of select food trees, particularly specimens with voluminous crowns capable of producing abundant fruit crops, appears to be key to the establishment and expansion of V. variegata groups. Our analysis of year-long fruit utilization revealed a high degree of preference for several species of trees. Two species exhibited mature fruit in a low percentage of stems but were available for a protracted period of time, while two additional species showed high intraspecific fruiting synchrony and were available for a shorter period of time. These contrasting phenologies, rather than the individual tree species, may be most important to V. variegata due to their coincident timing of fruit maturation with key lemur life-history events. Any disturbance-natural or anthropogenic-that disrupts the phenology cycles of food trees has the potential to impact lemur abundance and dispersion. Intense disturbances, such as heavy logging or severe cyclones, have long-lasting impacts on fruit production, while selective logging or moderate cyclonic windthrow cause more transient impacts. V. variegata is adapted to deal with an intrinsically erratic food supply by virtue of its reproductive biology and social behavior. PMID- 15898067 TI - Preliminary biomedical evaluation of wild ruffed lemurs (Varecia variegata and V. rubra). AB - Complete medical examinations were performed on 11 wild ruffed lemurs (Varecia variegata and V. rubra) from three sites in Madagascar. Each animal received a complete physical examination, several physiological parameters were analyzed (complete blood count, serum biochemical profile, and fecal bacterial culture), and the animals were examined for endo-, ecto-, and hemoparasites. Additional tests were performed as samples were available, including fat-soluble vitamin analysis, trace mineral analysis, toxoplasmosis serology, and viral serology. We found that the ruffed lemurs were in good health, harbored a low endoparasite load, and frequently had external parasites (e.g., ticks (Haemophysalis lemuris)). Statistically significant differences between captive and wild lemurs were found for the following serum biochemical and blood count parameters: alanine aminotransferase (ALT), total protein (TP), albumin, blood urea nitrogen, cholesterol, glucose, amylase, band neutrophil count, and eosinophil count. Low blood urea nitrogen (BUN) and serum cholesterol values in wild lemurs (compared to those of North American captive zoo ruffed lemurs) may suggest differences between diets in the wild and captivity. PMID- 15898068 TI - Environmental enrichment to address behavioral differences between wild and captive black-and-white ruffed lemurs (Varecia variegata). AB - I compared the behaviors of wild Varecia variegata living in a Malagasy rain forest with those of caged groups living in zoos in the United Kingdom in order to design environmental enrichment to encourage more natural behaviors. Comparisons were made between wild and captive animals in terms of activity budgets (instantaneously sampled at 1-min intervals) and social and solitary behaviors, which were continuously recorded for focal individuals. I followed the same sampling protocol during behavioral enrichment experiments, with additional monitoring of the amount and type of food consumed, and with more detailed observations of feeding behavior. No significant differences were found in resting or moving between wild and captive V. variegata. However, captive V. variegata spent more time on self-grooming and social behaviors, and less time feeding than wild V. variegata. There was also a lack of manual manipulation of food items. Behavioral enrichment experiments were carried out in which whole rather than chopped fruit was provided and presented in a more naturalistic manner. With this method of dietary presentation, manual manipulation of dietary items increased. Time spent feeding also increased significantly. Captive conservation breeding programs should not be wholly concerned with maintaining a diverse gene pool-they should also be concerned with conserving species-typical behaviors, especially if they are to produce behaviorally intact captive animals that can be reintroduced to the wild with minimal training, financial resources, and loss of individuals. PMID- 15898069 TI - Does female dominance facilitate feeding priority in black-and-white ruffed lemurs (Varecia variegata) in southeastern Madagascar? AB - Although many Malagasy lemurs are thought to be female dominant and to have female feeding priority, to date the relationship between these behaviors has been rigorously established only in Lemur catta, and other ways that females might achieve feeding priority have not been examined closely. Erhart and Overdorff [International Journal of Primatology 20:927-940, 1999] suggested that one way female primates achieve feeding priority is to initiate and lead groups to food, thereby gaining access to the food first and positively influencing their food intake compared to other group members. Here we describe female dominance patterns and potential measures of feeding priority in two groups of black-and-white ruffed lemurs (Varecia variegata) that were observed over a 15 month period in southeastern Madagascar. We predicted that the females would 1) be consistently dominant to males, 2) lead groups to food sources more often than males, and 3) have higher feeding rates compared to males when they arrived at food sources first. The results were dissimilar between the study groups. During the study, the oldest adult female in group 1 died. There was no evidence for female dominance in this group, and the remaining (likely natal) female did not lead the group more often, nor did she have a higher food intake than males. Group 1 dispersed shortly after the time frame reported here. In contrast, the resident female in group 2 was dominant to group males (based on agonistic interactions), led the group to food sources more often, and experienced a higher food intake when she arrived first at a food source. How these patterns vary over time and are influenced by the number of females in groups, group stability, food quality, and reproductive condition will be examined in future analyses. PMID- 15898070 TI - New developments in the behavioral ecology and conservation of ruffed lemurs (Varecia). AB - The papers in this issue were presented at a symposium during the 25th Annual Meeting of the American Society of Primatologists held in Oklahoma City, Oklahoma, in June 2002. This symposium brought together many of the scientists who have contributed to our knowledge of ruffed lemur ecology, behavior, and conservation in the past decade. One objective was to share and compare key findings about ruffed lemurs (Varecia) resulting from long-term field studies at various sites in Madagascar. A second objective was to cross-fertilize work being done in the wild with that being done in captivity, with the aim of advancing a common conservation mission for this critically endangered genus. Varecia is a prime candidate for synthetic assessments such as these because it has now been studied in both the northern and southern reaches of its geographic range, and has also been the focus of a captive-to-wild reinforcement project. The papers in this issue contribute to 1) the establishment of reference ranges for a suite of physiological parameters in healthy wild Varecia populations; 2) environmental enrichment aimed at preserving species-typical behaviors in captivity; 3) an understanding of how forest structure, floristic composition, and fruiting phenology in areas with differing disturbance histories correlate with the natural occurrence and abundance of Varecia; 4) primary knowledge concerning dominance relations between the sexes and group leadership in wild Varecia; and 5) primary knowledge concerning how wild Varecia, with their unusual reproductive pattern and heavy reliance on fruit, modulate their activity budgets seasonally and in tandem with reproductive stages. PMID- 15898071 TI - Activity budgets and activity rhythms in red ruffed lemurs (Varecia rubra) on the Masoala Peninsula, Madagascar: seasonality and reproductive energetics. AB - The activity budgets and daily activity rhythms of Varecia rubra were examined over an annual cycle according to season and reproductive stage. Given the relatively high reproductive costs and patchy food resources of this species, I predicted that V. rubra would 1) travel less and feed more during seasonal resource scarcity in an attempt to maintain energy balance, and 2) show sex differences in activity budgets due to differing reproductive investment. Contrary to the first prediction, V. rubra does not increase feeding time during seasonal food scarcity; rather, females feed for a consistent amount of time in every season, whereas males feed most during the resource-rich, hot dry season. The results are consistent with other predictions: V. rubra travels less in the resource-scarce cold rainy season, and there are some pronounced sex differences, with females feeding more and resting less than males in every season and in every reproductive stage except gestation. However, there are also some provocative similarities between the sexes when activity budgets are examined by reproductive stage. During gestation, female and male activity budgets do not differ and appear geared toward energy accumulation: both sexes feed and rest extensively and travel least during this stage. During lactation, activity budgets are geared toward high energy expenditure: both sexes travel most and in equal measure, and rest least, although it remains the case that females feed more and rest less than males. These similarities between female and male activity budgets appear related to cooperative infant care. The high energetic costs of reproduction in V. rubra females may require that they allot more time to feeding year round, and that their overall activity budget be more directly responsive to seasonal climate change, seasonal food distribution, and reproductive schedules. PMID- 15898072 TI - Chronic total occlusions in the superficial femoral artery: a novel strategy using a 1.5 mm J-tip hydrophilic guidewire with an over-the-wire balloon catheter under ultrasound guidance. AB - Chronic total occlusions in the superficial femoral artery (SFA) are the longest and straightest lesions in the whole body. This presents additional technical challenges and the inability to cross the occluded lesion is a common cause of procedural failure in these percutaneous interventions. The objective of this study was to investigate the usefulness of a strategy using a hydrophilic 1.5 mm J-tip 0.035'' guidewire with an over-the-wire balloon catheter under ultrasound guidance for chronic total occlusions in the SFA. This strategy was performed in 32 consecutive patients (36 limbs). Average occlusion length was 17 +/- 10 cm (3 40 cm). The crossover approach was performed in 26 cases (72%); in the remaining 10 cases, the antegrade ipsilateral approach was selected. The technical success rate was 92% (32/36). A secondary popliteal artery approach was performed in the three failed limbs and was successful in two limbs, increasing the final technical success rate to 97%. This novel procedure may be considered the preferred strategy for intervening in chronic total occlusions in the SFA. PMID- 15898073 TI - Expression profiling of genes crucial for placental and preimplantation development in bovine in vivo, in vitro, and nuclear transfer blastocysts. AB - Placental abnormalities and failed implantation are characterized phenotypes that occur in many species as a result of somatic cell cloning. This study examines a number of genes, critical for early placental development and reports aberrant expression patterns in a number of cloned bovine blastocysts, thus implicating a role of these genes in failed implantation. Messenger RNA (mRNA) expression of eight genes critical for early placental and preimplantation development including Acrogranin, Cdx2, Eomes, ErbB3, ERR2, Hand1, MRJ, and Rex1 were analyzed in single, in vivo, in vitro, and cloned bovine blastocysts (produced by hand-made cloning (HMC) and serial hand-made cloning (SHMC)) following complementary DNA (cDNA) amplification with a SMART cDNA synthesis kit. Aberrant expression of Acrogranin, Cdx2, and ERR2 was detected in a number of blastocysts produced by SHMC. Other genes, Eomes and Hand1, were not detectable in, in vivo bovine blastocysts, suggesting a differential expression pattern between bovine and murine embryos. A number of control marker genes including Oct4, IFN-tau, and PolyA were expressed in all single blastocysts analyzed. This is the first study to report that failure of implantation may be due to aberrant expression of genes in the preimplantation cloned embryo, which are crucial for the early regulation and differentiation of the placenta. PMID- 15898074 TI - Intrahepatic cholestasis: summary of an American Association for the Study of Liver Diseases single-topic conference. PMID- 15898075 TI - Identification of genes affecting lycopene accumulation in Escherichia coli using a shot-gun method. AB - Genes enhancing lycopene production in Escherichia coli were identified through colorimetric screening of shot-gun library clones constructed with E. coli chromosomal DNA. These E. coli cells had been engineered to produce lycopene, a red-colored carotenoid, which enabled screening for genes that enhance lycopene production. Six clones with enhanced lycopene production were isolated. Among 13 genes in these clones, dxs, appY, crl, and rpoS were found to be involved in enhanced lycopene production. While dxs and rpoS have been already reported to enhance lycopene production, appY and crl have not. DXP (1-deoxy-D-xylulose-5 phosphate) synthase is encoded by dxs and participates in the rate-limiting step in the synthesis of isopentenyl pyrophosphate (IPP), a building block of lycopene. Sigma S factor, encoded by rpoS, regulates transcription of genes induced at the stationary phase. The appY and crl genes encode transcriptional regulators related to anaerobic energy metabolism and the formation of curli surface fibers, respectively. E. coli harboring appY plasmids produced 2.8 mg lycopene/g dry cell weight (DCW), the same amount obtained with dxs despite the fact that appY is not directly involved in the lycopene synthesis pathway. The co expression of appY, crl, and rpoS with dxs synergistically enhanced lycopene production. The co-expression of appY with dxs produced eight times the amount of lycopene (4.7 mg/g DCW) that was produced without expression of both genes (0.6 mg/g DCW). PMID- 15898076 TI - Botulinum toxin-physiology and applications in head and neck disorders. AB - This article is structured around a literature review that was carried out using Ovid and Medline with the key words "botulinum," "toxin," and "ENT." Botulinum toxin has been used safely in humans for more than 20 years. The effects are transient, such that treatments are required to be repeated at intervals. Its application to ENT provides a useful tool to treat dystonia, autonomic dysfunction, facial nerve paresis, and hyperfunctional lines. It may also be of benefit in laryngeal rebalancing and the treatment of headaches. Further research is being carried out and new indications for treatment with botulinum toxin may include sialorrhea and rhinorrhea. PMID- 15898077 TI - Evaluation of computer-assisted instruction in histology: effect of interaction on learning outcome. AB - The purpose of this study was to determine better strategies for the design and use of computer-assisted instruction (CAI) in health science subjects that require visual learning. Evaluation of current use of CAI was focused on three CD based modules developed to teach histological images to beginning medical students at multiple sites. For internal control, students' learning outcomes and perceived effectiveness were analyzed with their demographic characteristics, computer attitude, computer experience, and learning behaviors being considered. Results indicated that students who used at least two different CAI programs scored significantly higher on the final examination than those who used only the CAI tool designed by their site's instructor. Further investigation indicated that students might have benefited from the interactive features of a specific CAI tool. Such scaffolds could have successfully supported encoding processes while students were restructuring their mental models. In addition, students perceived the CAI programs to be more effective when the tools were fully integrated into the curriculum. Perceived module effectiveness was significantly correlated with examination performance, suggesting a well-designed and appropriately used CAI tool may help students achieve not only learning efficiency, but also better learning outcome. PMID- 15898078 TI - A new way to teach an old topic: the cadaver-based anatomy short course for high school students. AB - An outreach program has been developed at the Department of Biological Sciences, Northern Illinois University (NIU), that offers an intensive short course in human gross anatomy to high school students and their teachers. Prior to the short course, the high school teachers attend a laboratory orientation, view the facilities, and then develop a syllabus suitable for the class needs. When the students arrive, they spend between 1 and 2 days rotating through five different work stations. Using their syllabus, they identify and learn the function of various structures on human cadavers and models. Evaluation and content testing of 143 students demonstrated a significant degree of enthusiasm and retention of the syllabus material at the end of the course. All teachers who participated in the program indicated that they wish to return the following year and new groups have already applied to attend. The results of the program demonstrate ways that universities can generate excitement about science and learning at the high school level. PMID- 15898079 TI - Promoting graduate student interest and participation in human gross anatomy. AB - For many years, graduate students at the University of North Texas Health Science Center (UNTHSC) were reluctant to enroll in dissection-based human gross anatomy courses. Furthermore, few graduate faculty mentors would allow their students to enroll in these courses. The significant amount of time allotted to courses such as anatomy and its effect on students' research programs have been identified by faculty as the primary reason for this lack of enthusiasm. For example, prior to 1999, graduate students taking human gross anatomy at UNTHSC registered for a 13 semester credit hour (SCH) course that was offered only in the fall semester. In the last 5 years, the anatomy teaching faculty in the Department of Cell Biology and Genetics (CGEN) restructured the human gross anatomy course for graduate students. A series of small, compact anatomy courses, ranging from 3-7 SCHs, are now offered throughout the school year to replace the single anatomy course. The CGEN faculty designed courses based on single or multiple body systems that varied in length from a few weeks to an entire semester. This change was initiated with the implementation of a system-based approach to anatomy instruction in our medical school curriculum and the elimination of our graduate anatomy course. With the development of six anatomy courses covering the entire human body, we have had a significant increase in graduate student participation. Moreover, the shorter duration of the courses has made them more appealing to graduate faculty mentors who want to keep graduate students focused on their research. PMID- 15898080 TI - Feulgen reaction study of novel threadlike structures (Bonghan ducts) on the surfaces of mammalian organs. AB - Threadlike structures on the surfaces of internal organs, which are thought to be part of the Bonghan duct system, were first reported about 40 years ago, but have been largely ignored since then. Recently, they were rediscovered, and in this study we discuss the Feulgen reaction that specifically stains DNA in order to identify these structures on the surface of rabbit livers as part of the Bonghan system. The distribution, shapes, and sizes of their nuclei are found to be similar to those of intravascular threadlike structures. The endothelial nuclei are rod-shaped, 10-20 mum long, and aligned in a broken-line striped fashion. The threadlike structure consists of a bundle of several subducts, which is a characteristic feature of Bonghan ducts and distinguishes them morphologically from lymphatic vessels. In addition, the Feulgen reaction clearly demonstrates that the subducts pass through a corpuscle, which is usually irregular or oval shaped and is connected to two or several threadlike structures that form a web on the surfaces of organs. Furthermore, spherical granules of about 1 mum in diameter are detected in the subducts. These granules were well stained by using the Feulgen reaction, which implies that they contain DNA. According to previous reports, a granule is a type of microcell and plays an essential role in the physiology and therapeutic effect of the Bonghan system and acupuncture. This role has yet to be elucidated. PMID- 15898082 TI - Provision of anatomical teaching in a new British medical school: getting the right mix. AB - In response to a government report, which recommended a substantial increase in the number of medical students in the United Kingdom by 2005, several new medical schools have been set up throughout the country. One such school, the Brighton and Sussex Medical School (BSMS), recently opened its doors to new students. BSMS offers a 5-year medical curriculum that uses an integrated systems-based approach to cultivate academic knowledge and clinical experience. Anatomy is one of the core elements of the program and, as such, features strongly within the modular curriculum. The challenge for the anatomy faculty has been to decide how best to integrate anatomy into the new curriculum and what teaching modalities should be used. A multidisciplinary approach has been taken using both traditional and contemporary teaching methods. Unlike most of the other new medical schools, BSMS uses cadaveric dissection as the cornerstone of its teaching, as the faculty believes that dissection still provides the most powerful technique for demonstrating anatomy as well as enhancing communication and teamwork skills. The dissection experience is handled using an understanding and professional way. However, to ensure that our students do not become detached from the process of patient-focused care, emphasis in the dissecting room environment is also placed on respect and compassion. To enhance conceptual understanding of structure and function and provide further clinical relevance, we are using imaging technology to demonstrate living anatomy. Unique to the BSMS curriculum is the teaching of the anatomy in the later years of the program. During specialist rotations, students will return to the dissecting room to study the anatomy relevant to that area. Such vertical integration ensures that core anatomical knowledge is gained at the most appropriate level relative to a student's clinical experience. PMID- 15898084 TI - Virtual laboratory manual for microscopic anatomy. AB - Using digital technology, we have assembled a virtual laboratory manual (VLM) that is a Web-based copy of our traditional laboratory manual. The VLM is used to enhance traditional laboratory instruction in histology. For each reference in the VLM to either a histological slide or an electron micrograph (EM), hyperlinks are included that download digital images derived from the students' glass slide sets or scanned EMs. The VLM serves as an atlas of digital images for concurrent study of similar sections by light microscopy during laboratory sessions. In addition, students can review the images from the VLM at remote locations. We have encouraged continued use of light microscopes in laboratories by basing the majority of practical examination identifications on analysis of marked histological slides that require students to use their own microscopes. The VLM provides the convenience of a Web-based resource with high-quality images, yet allows retention of the many excellent traditional aspects of our course. An example of a VLM laboratory on epithelium is available online (http://users.von.uc.edu/michaeje/VLM-Epithelium/exLab4.pdf). PMID- 15898085 TI - Deletion of N-terminus of human tyrosine hydroxylase type 1 enhances stability of the enzyme in AtT-20 cells. AB - Wildtype human tyrosine hydroxylase (TH) type 1 and 4 mutants (del-52, a form with the first 52 amino acid residues deleted; del-157, one with the first 157 amino acid residues deleted; RR-EE, one in which Arg37-Arg38 was replaced by Glu37-Glu38; and S40D, one in which Ser40 was replaced by Asp40) were expressed in AtT-20 mouse neuroendocrine cells in order to clarify how deeply the N terminus of TH is involved in the efficient production of dopamine (DA) in mammalian cells. The amounts of DA that accumulated in AtT-20 cells expressing these human TH type 1 (hTH1) phenotypes were in the following order: del-52 = del 157 = RR-EE > S40D > wildtype, although the enzyme activities of del-52 and del 157 were lower than those of wildtype, RR-EE, and S40D. The observation on immunoblot analyses that the N-terminus-deleted hTH1 mutants were much more stable than wildtype can reconcile the discrepant results. Computer-assisted analysis of the spatial configuration of hTH1 identified five newly recognized PEST motifs, one of which was located in the N-terminus sequence of Met1-Lys12 and predicted that deletion of the N-terminus region would alter the secondary structure within the catalytic domain. Collectively, the high stability of the N terminus-deleted hTH1 mutants can be generated by the loss of a PEST motif in their N-termini and the structural change in the catalytic domain, which would promise an efficient production of DA in mammalian cells expressing N-terminus deleted hTH1. PMID- 15898086 TI - Monitoring synaptogenesis in the developing mouse cerebellum with an original oligonucleotide microarray. AB - Use of DNA microarrays in neuroscience have been limited to rough screening purposes, mainly because the reliability and sensitivity of available arrays are not high enough. Because only a few hundred to one thousand genes are usually found to change expression levels in most experiments, we attempted to develop a more quantitative array by the following strategies: 1) limit the genes to those relevant to the experimental system, 2) design oligonucleotide probes of specified molecular properties so that more stringent hybridization conditions can be employed, 3) place six spots per probe on one slide and use multiple normalization genes, and 4) use a new type of gold-coated slide with higher S/N ratio. Genes involved in the construction and functioning of the synapse were selected from the literature as well as experimentally by comparing cerebella from hypothyroid and control mice at postnatal day 15 (P15). Although the number of genes covered was not yet large (172 genes), the custom array "Synaptoarray" thus constructed was capable of detecting +/-20% difference in gene expression ratios. Analysis of the postnatal development of the mouse cerebellum by using Synaptoarray demonstrated a general expression pattern with a peak at P7, followed by a decline at P15 and a partial recovery after P21. P10 clearly marked the end of the initial growth stage and a major transcriptional turning point in this system. This result suggests that such a custom array should be useful in monitoring perturbations to the normal developmental program. PMID- 15898087 TI - Self-reported symptoms of neurotoxicity and agricultural injuries among Ohio cash grain farmers. AB - BACKGROUND: In 1993, the Ohio Farm Family Health and Hazard Surveillance Program (OFFHHSP) was initiated to assess the health status of cash-grain farmers and their families. The objective of this study was to examine the postulated relationship between symptoms of neurotoxicity and risk of agriculture-related injury among Ohio cash-grain farmers. METHODS: A reanalysis of the data from a previously published case-control study of cash-grain farmers [Crawford et al. (1998) Am J Ind Med 34:588-599] was performed. The Principal Operator (PO) of each farm was asked to complete a self-administered questionnaire which included items about injuries experienced during the past year. A section of the questionnaire contained a 24-item instrument ("Q24") that queried symptoms of neurotoxicity experienced during the previous 12 months. Design-based multiple logistic regression analyses were conducted to evaluate associations between neuropsychological subsets ("domains") of the 24-item instrument and injury risk among the white male POs (who accounted for >99% of all POs). RESULTS: Significantly elevated odds ratios (ORs) were found in 8 of 11 domains. POs answering in the affirmative to the single item "Have you been bothered by lack of coordination or loss of balance?" were 3.12 times more likely to have reported an injury than POs responding negatively (95% CI 1.68-5.81). CONCLUSIONS: Farmers with higher scores on several Q24 domains, and for particular questionnaire items, appear to be at increased risk of agricultural injury. PMID- 15898088 TI - Acute methyl iodide exposure with delayed neuropsychiatric sequelae: report of a case. AB - BACKGROUND: Methyl iodide is a monohalomethane used as an analytic and organic chemistry reagent, as a methylating agent in organic chemical synthesis, and as a fumigant. In an acute exposure, methyl iodide is a pulmonary and dermal irritant. Chronic neurotoxicity has been reported in survivors of acute exposure. METHODS: A review of the 11 case reports of methyl iodide poisoning in the medical literature of the 20th century found that six of the patients experienced a chronic neurological syndrome characterized primarily by delayed psychiatric, behavioral, and cognitive sequelae. RESULTS: The case patient experienced a massive exposure to methyl iodide with resulting life-threatening burns. During convalescence, various cognitive and behavioral deficits became apparent. The results of a comprehensive evaluation at our occupational toxicology clinic, which included sequential neuropsychometric testing, are described. CONCLUSION: The findings in the case patient may advance our understanding of the mechanisms and clinical manifestations of chronic neurotoxicity after exposure to methyl iodide. PMID- 15898089 TI - Prevalence of asthma by industry in the US population: a study of 2001 NHIS data. AB - BACKGROUND: The estimated number of US workers potentially exposed to asthmagens ranges from 8 to 20 million. This study was undertaken to estimate the US prevalence of asthma in adults by industry of employment and to identify industries with elevated risk of asthma. METHODS: Prevalence analysis was performed on 20,991 adults, 18 years of age and older who participated in the 2001 National Health Interview survey. We used SUDAAN software to estimate the prevalence of self-reported physician diagnosed asthma by industry, and odds ratios (ORs) for asthma and industry adjusted for age, sex, race, and smoking status. RESULTS: The overall prevalence of physician diagnosed asthma was 6.5% (95% CI 6.1-6.9); 4.7% (95% CI 4.1-5.3) for males and 8.5% (95% CI 7.9-9.1) for females. In whites, the prevalence and ORs were significantly elevated for printing, publishing, and allied industries (OR = 2.4, 95% CI 1.2-5.0) and health care (OR = 1.3, 95% CI 1.0-1.7). In blacks, ORs were elevated for furniture, lumber, and wood (OR = 5.9, 95% CI 1.4-25.4) and entertainment and recreation industries (OR = 4.1, 95% CI 1.1-15.9). Other industries with elevated ORs included automobile dealers and gasoline station; durable goods; elementary, secondary schools, and colleges; other personal services; eating and drinking places; entertainment and recreation services; and utility and sanitary. CONCLUSIONS: Industries with elevated prevalence of asthma are identified. This information helps to target workplaces where detailed investigations for prevention and control may be appropriate. PMID- 15898090 TI - Classification and coding of commercial fishing injuries by work processes: an experience in the Danish fresh market fishing industry. AB - BACKGROUND: Work-related injuries in commercial fishing are of concern internationally. To better identify the causes of injury, this study coded occupational injuries by working processes in commercial fishing for fresh market fish. METHODS: A classification system of the work processes was developed by participation in fishing vessel trips where observations and video recordings of the work operations on board were collected. Subsequently the system was pilot tested using the Danish Maritime Authority injury reports. RESULTS: The developed classification system contains 17 main categories and up to 13 sub-categories of the work processes for each of the five different types of fishing. A total of 620 injury reports were reviewed and coded. Five percent (n = 33) of these were fatal injuries. The working processes were identified and coded according to the developed classification system for 553 (89%) injury reports: Danish seiner (n = 83), gill-netter (n = 122), beam trawler (n = 71), twin-trawler 2-T (n = 96), single/pair trawler 1-T (n = 181). Sixty-seven (11%) of the reports were unclassifiable due to lack of information. Preparing, shooting, and hauling of the gear and nets accounted for 50% of the injuries; they were most serious type of injuries such as fractures and sprains. Walking about the ship, in particular embarking and disembarking, climbing and descending ladders accounted for nearly one-fifth of the injuries. CONCLUSION: We found that the working processes related to working with the gear and nets vary greatly in the different fishing methods. Coding of the injuries to the specific working processes allows for targeted prevention efforts. PMID- 15898091 TI - Societal cost of workplace homicides in the United States, 1992-2001. AB - BACKGROUND: The Census of Fatal Occupational Injuries (CFOI) reported 8,672 workplace homicide victims between 1992 and 2001. Although rarely calculated, cost estimates are important for prevention and research efforts. METHODS: Societal costs were estimated using the cost-of-illness approach applied to CFOI data. The cost calculation model incorporated medical expenses, future earnings summed from the year of death until the year the decedent would have been 67, and household production losses (includes activities such as child care and housework). RESULTS: Workplace homicide had a total cost of nearly 6.5 billion dollars and a mean cost of 800,000 dollars between 1992 and 2001. The retail trade industry division had the highest number of homicides and total cost, 2.1 billion dollars, for males and 556,000 dollars for females. CONCLUSIONS: Estimates of the cost of work-related homicides can be used to improve occupational injury prevention and control program planning, policy analysis, evaluation of safety and health interventions, and advocacy for a safer work environment. PMID- 15898092 TI - Urinary metal and polycyclic aromatic hydrocarbon biomarkers in boilermakers exposed to metal fume and residual oil fly ash. AB - BACKGROUND: Boilermakers are occupationally exposed to known carcinogens. METHODS: The association of urinary 1-hydroxy-pyrene (1-OHP), a biomarker of polycyclic aromatic hydrocarbon (PAH) exposure, with biomarkers of metal exposure (vanadium, chromium, manganese, nickel, copper, and lead) in boilermakers exposed to metal fume from welding and dust particulates from residual oil fly ash (ROFA) was examined. A repeated measures cohort study was conducted during the overhaul of an oil-fired boiler. Twice-daily urine samples were obtained for 5 days and analyzed for cotinine, 1-OHP, and metals. Generalized estimating equations (GEE) were used to model the multivariate relationship of 1-OHP to the explanatory variables. RESULTS: Metal and 1-OHP levels were determined for 165 urine samples from 20 boilermakers and these levels increased during the workweek. However, the 1-OHP level was not significantly associated with any individual metal level at any time point. CONCLUSION: This suggests that boilermakers were occupationally exposed to PAH and metals, but 1-OHP as a PAH biomarker was unable to serve as a surrogate marker of metal exposure for the metals measured in this study. PMID- 15898093 TI - Acute and chronic health effects due to green tobacco exposure in agricultural workers. AB - BACKGROUND: An epidemiological study was undertaken in Gujarat, India to study the acute and chronic health effects of occupational exposure to green tobacco. METHODS: Non-Flue Cured Virginia (FCV) tobacco is the main crop in many districts of Central Gujarat. Three villages were selected from Anand district for the study and a random sample of 685 exposed workers were examined. Six hundred and fifty-five control workers with the same socio-economic status were examined from two villages where tobacco was not cultivated. RESULTS: The overall prevalence of green tobacco sickness (GTS) was 47.0% among tobacco workers. The prevalence in women workers was 55.7% while in men workers it was 42.66%. To detect the chronic health effects prevalence of hypertension, electrocardiogram (ECG) abnormalities, and eye problems in all the workers and reproductive abnormalities in women workers, all subjects received a medical examination. The data were compared in exposed and control group but they were non-significant statistically. No case of tobacco amblyopia was detected. CONCLUSION: The prevalence of GTS among non-FCV tobacco workers is high. However, from viewpoint of severity it can be considered as mild acute nicotine toxicity, which is relieved without medication. No significant difference is observed as regards to chronic health effects among tobacco workers and control workers. PMID- 15898094 TI - Mortality in Florida professional firefighters, 1972 to 1999. AB - BACKGROUND: Exposure to occupational hazards among firefighters may lead to increased mortality from cancer, lung, or heart disease. METHODS: Age- and gender adjusted mortality rates of 34,796 male and 2,017 female Florida professional firefighters between 1972 and 1999 were compared with the Florida general population. RESULTS: One thousand four hundred eleven male and 38 female firefighter deaths with known causes were identified. In male firefighters, mortality due to all causes and most non-malignant diseases was significantly less than expected. There was no excess overall mortality from cancer, but excesses existed for male breast cancer [standardized mortality ratio (SMR = 7.41; 95% confidence interval (CI): 1.99-18.96) and thyroid cancer (SMR = 4.82; 95% CI: 1.30-12.34)]. Mortality from bladder cancer was increased and approached statistical significance (SMR = 1.79; 95% CI: 0.98-3.00). Firefighters certified between 1972 and 1976 had excess mortality from bladder cancer (SMR = 1.95; 95% CI: 1.04-3.33). Female firefighters had similar morality patterns to Florida women except for atherosclerotic heart disease (SMR = 3.85; 95% CI: 1.66-7.58). CONCLUSIONS: Excess mortality risk from bladder cancer may be related to occupational exposure during firefighting. The thyroid cancer and breast cancer risk in males, as well as the excess risk of cardiovascular disease mortality noted in females warrant further investigation. PMID- 15898095 TI - Collegium Ramazzini call for an international ban on asbestos. PMID- 15898096 TI - Physical and mental health symptoms among NYC transit workers seven and one-half months after the WTC attacks. AB - BACKGROUND: On September 11, 2001, 600-800 New York City transit (NYCT) workers were working near the World Trade Center (WTC) Towers. After the disaster, employees reported physical and mental health symptoms related to the event. METHODS: Two hundred sixty-nine NYC transit employees were surveyed for mental and physical health symptoms 7(1/2) months after the WTC disaster. RESULTS: Workers in the dust cloud at the time of the WTC collapse had significantly higher risk of persistent lower respiratory (OR = 9.85; 95% CI: 2.24, 58.93) and mucous membrane (OR = 4.91; 95% CI: 1.53, 16.22) symptoms, depressive symptoms (OR = 2.48; 95% CI: 1.12, 5.51), and PTSD symptoms (OR = 2.91; 95% CI: 1.003, 8.16) compared to those not exposed to the dust cloud. Additional WTC exposures and potential confounders were also analyzed. CONCLUSIONS: Clinical follow up for physical and psychological health conditions should be provided for public transportation workers in the event of a catastrophic event. PMID- 15898097 TI - DNA damage in mice treated with sulfur dioxide by inhalation. AB - Sulfur dioxide (SO2) is a ubiquitous air pollutant produced by the burning of fossil fuels. In this study, single-cell gel electrophoresis (the Comet assay) was used to evaluate the DNA damage produced by inhalation exposure of mice to SO2. Male and female mice were housed in exposure chambers and treated with 14.00 +/- 1.25, 28.00 +/- 1.98, 56.00 +/- 3.11, and 112.00 +/- 3.69 mg/m3 SO2 for 6 hr/day for 7 days, while control groups were exposed to filtered air. Comet assays were performed on blood lymphocytes and cells from the brain, lung, liver, spleen, kidney, intestine, and testicles of the animals. SO2 caused significant, dose-dependent increases in DNA damage, as measured by Olive tail moment, in all the cell types analyzed from both sexes of mice. The results indicate that inhalation exposure to SO2 damages the DNA of multiple organs in addition to the lung, and suggests that this damage could result in mutation, cancer, and other diseases related to DNA damage. Further work will be required to understand the ultimate toxicological significance of this damage. These data also suggest that detecting DNA damage in blood lymphocytes, using the Comet assay, may serve as a useful tool for evaluating the impact of pulmonary SO2 exposure in human biomonitoring studies. PMID- 15898098 TI - Efficacy of the 1.5 millisecond pulse-duration, 585 nm, pulsed-dye laser for treating port-wine stains. AB - BACKGROUND AND OBJECTIVES: Some reports have suggested that 1.5-millisecond pulse duration pulsed-dye lasers (PDLs) may have increased efficacy in treating port wine stains (PWSs) as compared to previous generation lasers. We study the efficacy of the 1.5-millisecond pulse-duration PDL for treating PWSs. STUDY DESIGN/MATERIALS AND METHODS: We treated 104 areas of involvement by PWSs in 95 subjects with the 1.5-millisecond pulse-duration PDL. Subjective and objective measures of improvement were utilized. Twenty-one of these PWSs had failed treatment with earlier generation PDLs. RESULTS: PWSs previously refractory to the shorter pulse duration lasers responded well to treatment, as did PWSs not previously treated with any laser. CONCLUSIONS: The 1.5-millisecond pulse duration PDL is effective for treating PWSs and may offer improved efficacy over shorter pulse-duration lasers. PMID- 15898099 TI - Peroxisomal participation in psychosine-mediated toxicity: implications for Krabbe's disease. AB - Psychosine (galactosylsphingosine) accumulation in globoid cell leukodystrophy (Krabbe's disease) results in the loss of myelin and oligodendrocytes. To understand the role of psychosine toxicity in Krabbe's disease, we examined the effects of psychosine on peroxisomal functions and their relationship with reactive oxygen species. Rat C(6) glial cells were treated with psychosine with and without cytokines. Peroxisomal beta-oxidation was significantly inhibited and very long chain fatty acid levels and free radicals were increased in treated cells. Furthermore, psychosine treatment decreased glutathione and ATP levels, plasmalogen content, and expression of alkyl-DHAP synthase. Brain tissue of twitcher mice (animal model of Krabbe's) had decreased beta-oxidation activity, low glutathione, and reduced plasmalogens. Psychosine treatment of rat primary oligodendrocytes inhibited peroxisomal activities. Psychosine-mediated loss of peroxisomal function and free radical production was inhibited with the antioxidant N-acetylcysteine in glial cells. Our results suggest that inhibition of peroxisomal functions and increased free radical production by psychosine may be partly responsible for oligodendrocyte and myelin loss observed in the Krabbe's brain, and that antioxidant therapy may be useful in the treatment of Krabbe's disease. PMID- 15898100 TI - Hyperosmotic mannitol induces Src kinase-dependent phosphorylation of beta catenin in cerebral endothelial cells. AB - Mannitol, which is a cell-impermeable and nontoxic polyalcohol, has been shown to be a useful tool for reversible opening of the blood-brain barrier (BBB). Despite successful clinical trials, the molecular mechanism of the mannitol-induced changes in cerebral endothelial cells (CECs) are poorly understood. For our experiments, we used CECs in culture, which were treated with different, clinically relevant concentrations of mannitol. We found that mannitol induced a rapid, concentration-dependent, and reversible tyrosine phosphorylation of a broad range of proteins between 50 and 190 kDa. One of the targets of tyrosine phosphorylation turned out to be the adherens junction protein beta-catenin. Phosphorylation of beta-catenin on tyrosine residues caused its subcellular redistribution and its dissociation from cadherin and alpha-catenin as shown by coimmunoprecipitation studies. All these effects could be inhibited by the Src kinase inhibitor PP-1 but not by the Erk inhibitor U0126, the Rho kinase inhibitor Y27632, or the calcium channel blocker verapamil. Because beta-catenin is a key component of the junctional complex, its Src-mediated phpsphorylation may play an important role in the mannitol induced reversible opening of the BBB. PMID- 15898101 TI - CO(2) laser surgery in the treatment of glottic cancer. AB - BACKGROUND: The aim of the study was to assess the effectiveness of CO(2) laser endoscopic surgery in the treatment of glottic carcinoma limited to the true vocal cords or involving the adjacent regions. METHODS: Seven hundred nineteen patients (687 men and 32 women; mean age, 60.4 years; range, 33-86 years) with glottic carcinoma (432 T1N0M0, 236 T2N0M0, 51 T3N0M0) underwent CO(2) laser surgery (mean follow-up, 5 years; range, 2-17 years). Statistical comparison was carried out with Wilcoxon test, considering p < .05 the minimum significance value. RESULTS: Overall actuarial survival, adjusted actuarial survival, and percentage of patients with no evidence of disease at 5 years were 85%, 97%, and 85%, respectively, in patients with T1a disease; 84%, 96%, and 83% in those with T1b disease; 77%, 86%, and 61% in those with T2 unilateral tumors; 77%, 88%, and 55% in those with T2 bilateral tumors; and 64%, 72%, and 60% in those with T3 disease. The statistical analysis showed the following: significant differences in the comparison of T1 versus T2 and T2 versus T3 tumors (p < .01), with the exception of no evidence of disease in the comparison of T2 versus T3 (p > .05); and no significant differences in the comparison of unilateral and bilateral tumors (p > .05). Actuarial local control, actuarial nodal control, and actuarial distant metastasis control at 5 years were 85%, 98%, and 99%, respectively, in patients with T1 disease; and 66%, 82%, and 91% in patients with T2 disease; and 66%, 83%, and 95% in patients with T3 disease. The laryngeal preservation rate was 97.3% in the T1 group, 82.5% in the T2 group, and 80.5% in T3 group. CONCLUSIONS: CO(2) laser endoscopic surgery is effective in the treatment of glottic carcinoma not infiltrating the cartilaginous skeleton; the results achieved are competitive with those of open conservative operations, if we take into account the possibilities afforded by salvage surgery and the rate of laryngeal preservation achieved in the study patients. PMID- 15898102 TI - Differentiation of immature oligodendrocytes is regulated by phosphorylation of cyclic AMP-response element binding protein by a protein kinase C signaling cascade. AB - Previous experiments showed that the expression and phosphorylation levels of cyclic AMP-response element binding protein (CREB) are important factors that regulate oligodendrocyte differentiation. The present study was designed to determine whether CREB phosphorylation advances oligodendrocyte differentiation or vice versa and to identify the protein kinase that primarily regulates CREB phosphorylation. We examined the expression and phosphorylation levels of CREB in developing oligodendrocytes at a specific differentiation stage by double immunocytochemical staining with specific differentiation markers and antibody for phosphorylated CREB. We found that the CREB expression level increased along oligodendrocyte differentiation, and that its phosphorylated level was highest in immature oligodendrocytes. We also showed that CREB phosphorylation was regulated principally by protein kinase C (PKC) activity in immature oligodendrocytes. Our findings suggest that CREB phosphorylation is dependent on a PKC signaling cascade, and this phosphorylation activates CREB-mediated transcription and advances the differentiation of immature to mature oligodendrocytes. PMID- 15898104 TI - Endothelin increases expression of exon III- and exon IV-containing brain-derived neurotrophic factor transcripts in cultured astrocytes and rat brain. AB - The effects of endothelins (ETs) on brain-derived neurotrophic factor (BDNF) production in astrocytes were investigated. ET-1 (100 nM) increased the mRNA level and extracellular release of BDNF in cultured astrocytes. RT-PCR analyses using primer pairs that amplified exon-specific BDNF transcripts revealed that exon III- and exon IV-containing BDNF transcripts existed in cultured astrocytes, whereas exon I- and exon II-containing BDNF transcripts did not. ET-1 and Ala(1,3,11,15)-ET-1, an ET(B) receptor agonist, increased the expressions of the exon III and exon IV transcripts in cultured astrocytes. Intracerebroventricular administration of 500 pmol/day of Ala(1,3,11,15)-ET-1 increased exon III and exon IV BDNF transcripts in the rat striatum. In cultured astrocytes, Ca(2+) chelation, W-7 (a calmodulin inhibitor), and KN93 (a Ca(2+)/calmodulin kinase inhibitor) inhibited the increases in exon IV BDNF mRNA and CCAAT enhancer binding protein beta (C/EBPbeta) levels induced by ET-1. The ET-induced increases in exon III BDNF mRNA expression and phosphorylation of cAMP response element binding protein (CREB) were reduced by Ca(2+) chelation, W-7, KN93, PD98059 (a MEK inhibitor), and wortmannin (a phosphatidylinositol 3-kinase inhibitor). These results suggest that ETs stimulate the expressions of exon III and exon IV BDNF transcripts in astrocytes through CREB and C/EBPbeta-mediated mechanisms, respectively. PMID- 15898103 TI - Connexin43, the major gap junction protein of astrocytes, is down-regulated in inflamed white matter in an animal model of multiple sclerosis. AB - Both multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE), its animal model, involve inflammatory attack on central nervous system (CNS) white matter, leading to demyelination and axonal damage. Changes in astrocytic morphology and function are also prominent features of MS and EAE. Resting astrocytes form a network that is interconnected through gap junctions, composed mainly of connexin43 (Cx43) protein. Although astrocytic gap junctional connectivity is known to be altered in many CNS pathologies, little is known about Cx43 expression in inflammatory demyelinating disease. Therefore, we evaluated the expression of Cx43 in spinal cords of EAE mice compared with healthy controls. Lumbar ventral white matter areas were heavily infiltrated with CD11beta-immunoreactive monocytes, and within these infiltrated regions loss of Cx43 immunoreactivity was evident. These regions also showed axonal dystrophy, demonstrated by the abnormally dephosphorylated heavy-chain neurofilament proteins. Astrocytes in these Cx43-depleted lesions were strongly glial fibrillary acidic protein reactive. Significant loss (38%) of Cx43 protein in EAE mouse at the lumbar portion of spinal cords was confirmed by Western blot analysis. Decreased Cx43 transcript level was also observed on cDNA microarray analysis. In addition to changes in Cx43 expression, numerous other genes were altered, including those encoding adhesion and extracellular matrix proteins. Our data support the notion that, in addition to damage of myelinating glia, altered astrocyte connectivity is a prominent feature of inflammatory demyelination. PMID- 15898105 TI - Amino acid architecture and the distribution of polar atoms on the surfaces of proteins. AB - We propose that a necessary condition for a protein to be soluble is the absence of large hydrophobic patches on its solvent-accessible surface, which can cause aggregation to occur. We note that the polar nature of the backbone of all amino acids guarantees a minimum polar content and hence can interrupt such patches. As a result, a carefully conserved detailed atomic placement of residues on the protein surface is not necessary for solubility. In order to demonstrate this, we construct a measure based on the average hydrophobicity of a simply defined patch. We use this measurement to compare surfaces that exhibit a clear difference in their solubility properties, namely, a) the solvent accessible surfaces for a set of homo-dimers and the surfaces buried in their interfaces and b) for a set of monomers the surfaces of fragments of secondary structure which are solvent accessible/inaccessible. Having demonstrated a difference in the first set of distributions, we characterize the solvent accessible surfaces of monomeric proteins. To test if cooperative behavior occurs between the atoms for these surfaces, we construct a set of randomized surfaces, which obey a very simple stereochemical constraint. We find that the observed and randomized distributions are much more similar than the previous sets we examined. This implies that while surfaces of soluble proteins must have sufficient polar content, the relative placement of atoms of one amino acid with respect to the atoms of neighboring amino acid need not be finely tuned, which provides an innate robustness for protein design and folding. PMID- 15898106 TI - Homocysteic acid induces intraneuronal accumulation of neurotoxic Abeta42: implications for the pathogenesis of Alzheimer's disease. AB - The causes of neuronal dysfunction and degeneration in Alzheimer's disease (AD) are not fully understood, but increased production of neurotoxic forms of amyloid beta-peptide-42 (Abeta42) seems of major importance. Large extracellular deposits of aggregated Abeta42 (plaques) is a diagnostic feature of AD, but Abeta42 may be particularly cytotoxic when it accumulates inside neurons. The factors that may promote the intracellular accumulation of Abeta42 in AD are unknown, but recent findings suggest that individuals with elevated homocysteine levels are at increased risk for AD. We show that homocysteic acid (HA), an oxidized metabolite of homocysteine, induces intraneuronal accumulation of a Abeta42 that is associated with cytotoxicity. The neurotoxicity of HA can be attenuated by an inhibitor of gamma-secretase, the enzyme activity that generates Abeta42, suggesting a key role for intracellular Abeta42 accumulation in the neurotoxic action of HA. Concentrations of HA in cerebrospinal fluid (CSF) were similar in AD and control subjects. CSF homocysteine levels were elevated significantly in AD patients, however, and homocysteine exacerbated HA-induced neurotoxicity, suggesting a role for HA in the pathogenic action of elevated homocysteine levels in AD. These findings suggest that the intracellular accumulation of Abeta42 plays a role in the neurotoxic action of HA, and suggest a potential therapeutic benefit of agents that modify the production and neurotoxic actions of HA and homocysteine. PMID- 15898107 TI - Strategies for increasing the efficiency of a genetic algorithm for the structural optimization of nanoalloy clusters. AB - An improved genetic algorithm (GA) is described that has been developed to increase the efficiency of finding the global minimum energy isomers for nanoalloy clusters. The GA is optimized for the example Pt12Pd12, with specific investigation of: the effect of biasing the initial population by seeding; the effect of removing specified clusters from the population ("predation"); and the effect of varying the type of mutation operator applied. These changes are found to significantly enhance the efficiency of the GA, which is subsequently demonstrated by the application of the best strategy to a new cluster, namely Pt19Pd19. PMID- 15898108 TI - Role of the surface heterogeneity in adsorption of hydrogen ions on metal oxides: theory and simulations. AB - In this article we study the effect of energetic heterogeneity of a crystalline surface on the adsorption of hydrogen ions (protons) from the liquid phase. In particular, we examine the influence of the shape of the adsorption energy distribution on the equilibrium isotherms of hydrogen ions. To that purpose, a few popular distribution functions, including rectangular, exponential, and asymmetric Gaussian are considered. Additionally, multimodal distribution functions, which may correspond to the adsorption on different crystal planes of the oxide, are also used. Lateral interactions between adsorbed charges are modeled using the potential function proposed by Borkovec et al., which accounts also for polarization of the liquid medium. The results presented here are obtained using both Monte Carlo (MC) simulations and theoretical calculations involving Mean Field Approximation (MFA). They indicate that increased energetic heterogeneity of the adsorbing surface may, in general, considerably change the behavior of the adsorption isotherms, regardless of the assumed distribution function. It is also shown that the predictions of the proposed theory are consistent with the data obtained from the MC simulations. PMID- 15898109 TI - Practical conversion from torsion space to Cartesian space for in silico protein synthesis. AB - Many applications require a method for translating a large list of bond angles and bond lengths to precise atomic Cartesian coordinates. This simple but computationally consuming task occurs ubiquitously in modeling proteins, DNA, and other polymers as well as in many other fields such as robotics. To find an optimal method, algorithms can be compared by a number of operations, speed, intrinsic numerical stability, and parallelization. We discuss five established methods for growing a protein backbone by serial chain extension from bond angles and bond lengths. We introduce the Natural Extension Reference Frame (NeRF) method developed for Rosetta's chain extension subroutine, as well as an improved implementation. In comparison to traditional two-step rotations, vector algebra, or Quaternion product algorithms, the NeRF algorithm is superior for this application: it requires 47% fewer floating point operations, demonstrates the best intrinsic numerical stability, and offers prospects for parallel processor acceleration. The NeRF formalism factors the mathematical operations of chain extension into two independent terms with orthogonal subsets of the dependent variables; the apparent irreducibility of these factors hint that the minimal operation set may have been identified. Benchmarks are made on Intel Pentium and Motorola PowerPC CPUs. PMID- 15898110 TI - A combined freeze-and-cut strategy for the description of large molecular systems using a localized orbitals approach. AB - A technique to reduce the computational effort in calculating ab initio energies using a localized orbitals approach is presented. By exploiting freeze strategy at the self-consistent field (SCF) level and a cut of the unneeded atomic orbitals, it is possible to perform a localized complete active space (CAS-SCF) calculation on a reduced system. This will open the possibility to perform ab initio treatments on very large molecular systems, provided that the chemically important phenomena happen in a localized zone of the molecule. Two test cases are discussed, to illustrate the performance of the method: the cis-trans interconversion curves for the (7Z)-13 ammoniotridec-7-enoate, which demonstrates the ability of the method to reproduce the interactions between charged groups; and the cisoid-transoid energy barrier for the aldehydic group in the C13 polyenal molecule. PMID- 15898111 TI - Increased chromosome instability but not cancer predisposition in haploinsufficient Bub3 mice. AB - Mitotic spindle checkpoint proteins have been shown to play a crucial role in the accurate segregation of chromosomes during cell division. Bub3 is a member of a group of mitotic checkpoint proteins that are essential for this process. To investigate the role of Bub3 in chromosome segregation and cancer development, we analyzed haploinsufficient cells in mice. Heterozygous Bub3 embryonic fibroblasts displayed increased aneuploidy and premature sister-chromatid separation. In addition, when challenged with the microtubule disruptor nocodazole, the cells showed a slight increase in chromatid breakage and a decrease in the mitotic index. No substantial differences were observed between wild-type and Bub3 heterozygous mice in the frequency or the rate at which tumors appeared. Crossing Bub3(+/-) mice onto a heterozygous tumor-suppressor background of Trp53 or Rb1 similarly revealed no substantial differences in either the number or the rate at which tumors appeared. These results suggest that haploinsufficiency of Bub3 causes a slight increase in chromosome instability but is not clearly associated with a noticeable rise in the probability of tumor formation in the animal, possibly because of a partially functional mitotic checkpoint, or cells exhibiting chromosome instability could have activated the apoptosis pathway and thus escaped tumor induction and detection. PMID- 15898112 TI - A CSOV study of the difference between HF and DFT intermolecular interaction energy values: the importance of the charge transfer contribution. AB - Intermolecular interaction energy decompositions using the Constrained Space Orbital Variation (CSOV) method are carried out at the Hartree-Fock level on the one hand and using DFT with usual GGA functionals on the other for a number of model complexes to analyze the role of electron correlation in the intermolecular stabilization energy. In addition to the overall stabilization, the results provide information on the variation, with respect to the computational level, of the different contributions to the interaction energy. The complexes studied are the water linear dimer, the N-methylformamide dimer, the nucleic acid base pairs, the benzene-methane and benzene-N2 van der Waals complexes, [Cu+ -(ImH)3]2, where "ImH" stands for the Imidazole ligand, and ImH-Zn++. The variation of the frozen core energy (the sum of the intermolecular electrostatic energy and the Pauli repulsion energy) calculated from the unperturbed orbitals of the interacting entities indicates that the intramolecular correlation contributions can be stabilizing as well as destabilizing, and that general trends can be derived from the results obtained using usual density functionals. The most important difference between the values obtained from HF and DFT computations concerns the charge transfer contribution, which, in most cases, undergoes the largest increase. The physical meaning of these results is discussed. The present work gives reference calculations that might be used to parametrize new correlated molecular mechanics potentials. PMID- 15898113 TI - Phenoloxidase in larvae of Plodia interpunctella (Lepidoptera: Pyralidae): molecular cloning of the proenzyme cDNA and enzyme activity in larvae paralyzed and parasitized by Habrobracon hebetor (Hymenoptera: Braconidae). AB - Phenoloxidase (PO) is a major component of the insect immune system. The enzyme is involved in encapsulation and melanization processes as well as wound healing and cuticle sclerotization. PO is present as an inactive proenzyme, prophenoloxidase (PPO), which is activated via a protease cascade. In this study, we have cloned a full-length PPO1 cDNA and a partial PPO2 cDNA from the Indianmeal moth, Plodia interpunctella (Hubner) (Lepidoptera: Pyralidae) and documented changes in PO activity in larvae paralyzed and parasitized by the ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae). The cDNA for PPO1 is 2,748 bp and encodes a protein of 681 amino acids with a calculated molecular weight of 78,328 and pI of 6.41 containing a conserved proteolytic cleavage site found in other PPOs. P. interpunctella PPO1 ranges from 71-78% identical to other known lepidopteran PPO-1 sequences. Percent identity decreases as comparisons are made to PPO-1 of more divergent species in the orders Diptera (Aa-48; As-49; and Sb-60%) and Coleoptera (Tm-58; Hd-50%). Paralyzation of host larvae of P. interpunctella by the idiobiont H. hebetor results in an increase in phenoloxidase activity in host hemolymph, a process that may protect the host from microbial infection during self-provisioning by this wasp. Subsequent parasitization by H. hebetor larvae causes a decrease in hemolymph PO activity, which suggests that the larval parasitoid may be secreting an immunosuppressant into the host larva during feeding. PMID- 15898114 TI - Lipid analysis of the sex pheromone gland of the moth Heliothis virescens. AB - The sex pheromone gland of female Heliothis virescens was analyzed for fatty acid and lipid content. Base methanolysis of the gland showed a large amount of methyl (Z)-11-hexadecenoate (Z11-16:Acyl), the fatty acyl analog of the major pheromone component, (Z)-11-hexadecenal, as well as a small amount of methyl (Z)-11 octadecenoate. Methyl esters of various common fatty acids were also observed. HPTLC analysis of the glandular lipids revealed large quantities of triacylglycerols (TGs), and lesser amounts of 1,2-diacylglycerols (1,2-DGs), 2 monoacylglycerols (2-MGs), phosphatidyl ethanolamines, and phosphatidyl cholines. The greatest amount of Z11-16:Acyl in these lipids was in the TGs, with lesser amounts in the two phospholipid classes and only trace amounts in the other neutral lipids. The glands of females at various ages and photoperiodic times were extracted, fractionated into neutral and polar fractions by silica SPE, and fatty acid titers in these fractions determined. All fatty acids, but notably Z11 16:Acyl, showed significant total and neutral lipid fraction peaks at mid scotophase for 2-day-old females; a less dramatic, but significant, Z11-16:Acyl peak in the polar fraction was also observed. However, only a relatively small proportion (<50%) of this acid was recovered from the silica at all times. This "non-recoverable" Z11-16:Acyl showed a dramatic and significant peak at mid scotophase for 2-day females, corresponding roughly with maximal pheromone titer. All other acids in the gland were recovered in high proportions, and their respective "non-recoverable" titers were not different at any of the times analyzed. Based on previous work, this non-recoverable Z11-16:Acyl is likely the CoA ester. Therefore, it appears that the pheromone gland of H. virescens maintains pools of Z11-16:Acyl in both CoA ester and TG forms, which are available for biosynthesis of pheromone. These pools are greatest during maximal pheromone production when the biosynthetic enzymes, possibly the fatty acid reductase, are unable to utilize rapidly enough the quantities of Z11-16:Acyl biosynthesized. PMID- 15898115 TI - Molecular dynamics of detoxification and toxin-tolerance genes in brown planthopper (Nilaparvata lugens Stal., Homoptera: Delphacidae) feeding on resistant rice plants. AB - To investigate the molecular response of brown planthopper, Nilaparvata lugens (BPH) to BPH-resistant rice plants, we isolated cDNA fragments of the genes encoding for carboxylesterase (CAR), trypsin (TRY), cytochrome P450 monooxygenase (P450), NADH-quinone oxidoreductase (NQO), acetylcholinesterase (ACE), and Glutathione S-transferase (GST). Expression profiles of the genes were monitored on fourth instar nymphs feeding on rice varieties with different resistance levels. Northern blot hybridization showed that, compared with BPH reared on susceptible rice TN1, expression of the genes for P450 and CAR was apparently up regulated and TRY mRNA decreased in BPH feeding on a highly resistant rice line B5 and a moderately resistant rice variety MH63, respectively. Two transcripts of GST increased in BPH feeding on B5; but in BPH feeding on MH63, this gene was inducible and its expression reached a maximum level at 24 h, and then decreased slightly. The expression of NQO gene was enhanced in BPH on B5 plants but showed a constant expression in BPH on MH63 plants. No difference in ACE gene expression among BPH on different rice plants was detected by the RT-PCR method. The results suggest these genes may play important roles in the defense response of BPH to resistant rice. PMID- 15898116 TI - Characteristics of two genes encoding proteins with an ADAM-type metalloprotease domain, which are induced during the molting periods in Bombyx mori. AB - By microarray analyses, we identified two genes (BmADAMTS-1 and BmADAMTS-like) encoding a protein, which are induced during the pupal ecdysis in the wing discs of Bombyx mori; these genes are homologous to ADAMTS family members (a disintegrin and metalloproteinase domain, with thrombospondin type-1 repeats). A complete metal-binding motif of the ADAM-type metalloprotease domain (HEXXHXXGXXHD) was contained in both amino acid sequences. However, thrombospondin type 1 (TSP-1) repeats were observed only in BmADAMTS-1. The BmADAMTS-1 gene was expressed in the hemocyte and midgut of the larvae at day 2 of wandering stage (W2), and strongly induced during the pupal ecdysis in the hemolymph. The BmADAMTS-like gene was expressed in the epithelial tissues of the larvae at W2, and had expression peaks slightly later than the BmADAMTS-1 gene. Our results indicate that BmADAMTS-1 and BmADAMTS-like may cleave the extracellular matrix (ECM) in the degenerating and remodeling tissues during the molting periods. PMID- 15898117 TI - Comparison of cytocompatibility of zein film with other biomaterials and its degradability in vitro. AB - Cytocompatibility of particle zein (Pzein) and film zein (Fzein) was evaluated and compared with polyhydroxybutyrate (PHB), its copolymer poly(hydroxybutyrate co-hydroxyvalerate) (PHBV), polylactic acid (PLA), and collagen, using HL-7702 cells, in terms of cell attachment rate within 3 h, and cell viabilities at 3 and 6 days determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method. The zein degradation test was carried out using collagenase and trypsin, and the degradation product was added to the culture medium at different concentrations in order to examine the concentration-dependent cytotoxic effect. RESULT: The adhesion rate of the HL-7702 cells on both Pzein and Fzein was higher than that on collagen film. Cell viabilities were higher on both Pzein and Fzein than on films of PLA, PHB, PHBV, and collagen from fish skin. Zein can be degraded by both trypsin and collagenase, and the degradation product can enhance cell viability within a certain range of concentrations. PMID- 15898118 TI - Pheromone biosynthetic pathways in the moths Heliothis subflexa and Heliothis virescens. AB - Sex pheromones of many moth species have relatively simple structures consisting of a hydrocarbon chain with a functional group and one to several double bonds. These sex pheromones are derived from fatty acids through specific biosynthetic pathways. We investigated the incorporation of deuterium-labeled tetradecanoic, hexadecanoic, and octadecanoic acid precursors into pheromone components of Heliothis subflexa and Heliothis virescens. The two species utilize (Z)11 hexadecenal as the major pheromone component, which is produced by Delta11 desaturation of hexadecanoic acid. H. subflexa also produced (Z)11-hexadecanol and (Z)-11-hexadecenyl acetate via Delta11 desaturation. In H. subflexa, octadecanoic acid was used to biosynthesize the minor pheromone components (Z)9 hexadecenal, (Z)9-hexadecenol, and (Z)9-hexadecenyl acetate. These minor components are produced by Delta11 desaturation of octadecanoic acid followed by one round of chain-shortening. In contrast, H. virescens used hexadecanoic acid as a substrate to form (Z)11-hexadecenal and (Z)11-hexadecenol and hexadecenal. H. virescens also produced (Z)9-tetradecenal by Delta11 desaturation of the hexadecanoic acid followed by one round of chain-shortening and reduction. Tetradecanoic acid was not utilized as a precursor to form Z9-14:Ald in H. virescens. This labeling pattern indicates that the Delta11 desaturase is the only active desaturase present in the pheromone gland cells of both species. PMID- 15898119 TI - Physical activity and the risk of ovarian cancer: a case-control study in Canada. AB - We evaluated the impact of recreational and occupational physical activity on ovarian cancer risk using data from a population-based case-control study of 442 cases with histologically confirmed incident ovarian cancer and 2,135 controls aged 20-76 years, conducted in 1994-1997 in Canada. Frequency and intensity of physical activity were collected through self-administered questionnaires. Compared to women in the lowest tertiles of moderate, vigorous and total recreational activity, those in the highest tertiles had multivariable-adjusted odds ratios (and 95% confidence intervals) of 0.67 (0.50-0.88), 0.93 (0.70-1.24) and 0.73 (0.58-0.98), respectively. There were statistically significant trends of decreasing risk with increasing levels of moderate and total recreational activity, with similar patterns for premenopausal and postmenopausal women. A significant reduction in risk associated with higher level of moderate recreational activity was observed for serous, endometrioid and other but not mucinous types of tumors. The analyses in one province with the largest number of cases and controls indicated that occupational activity was associated with reduced ovarian cancer risk by lifetime activity and by various life periods (early 20s, early 30s, early 50s and 2 years before interview). Our study suggests that occupational and regular moderate recreational physical activity reduce ovarian cancer risk. PMID- 15898120 TI - Study of tumor cell invasion by Fourier transform infrared microspectroscopy. AB - Lung cancer is usually fatal once it becomes metastatic. However, in order to develop metastases, a tumor usually invades the basal membrane and enters the vascular or lymphatic system. In this study, a three-dimensional artificial membrane using collagen type I, one of the main components of basal membranes, was established in order to investigate tumor cell invasion. Lung cancer cell line CALU-1 was seeded on this artificial membrane and cell invasion was studied using the Fourier transform infrared (FTIR) imaging technique. This approach allowed identification of tumor cells invading the collagen type I membrane by means of their infrared spectra and images. The mapping images obtained with FTIR microspectroscopy were validated with standard histological section analysis. The FTIR image produced using a single wavenumber at 1080 cm(-1), corresponding to PO2- groups in DNA from cells, correlated well with the histological section, which clearly revealed a cell layer and invading cells within the membrane. Furthermore, the peaks corresponding to amide A, I, and II in the spectra of the invading cells shifted compared to the noninvading cells, which may relate to the changes in conformation and/or heterogeneity in the phenotype of the cells. The data presented in this study demonstrate that FTIR microspectroscopy can be a fast and reliable technique to assess tumor invasion in vitro. PMID- 15898121 TI - Rectal intussusception in symptomatic patients is different from that in asymptomatic volunteers. AB - BACKGROUND: Rectal intussusception is a common finding at evacuation proctography in both symptomatic and asymptomatic individuals. Little information exists, however, as to whether intussusception morphology differs between patients with evacuatory dysfunction and healthy volunteers. METHODS: Thirty patients (19 women; median age 44 (range 21-76) years) with disordered rectal evacuation, in whom an isolated intussusception was seen on proctography, were studied. Various morphological parameters were measured, and compared with those from 11 asymptomatic controls (six women; median age 30 (range 24-38) years) found, from 31 volunteers, to have rectal intussusception. Intussusceptum thickness greater than 3 mm was designated as full thickness. Intussuscepta impeding evacuation were deemed to be occluding. RESULTS: Twenty-two patients had full-thickness intussusception, compared with two controls (P = 0.003). Intussusceptum thickness was significantly greater in the symptomatic group (anterior component: P = 0.004; posterior: P = 0.011). Twenty patients in the symptomatic group, but only three subjects in the control group, had a mechanically occluding intussusception (P = 0.043), although only three patients demonstrated evacuatory dynamics outside the normal range. CONCLUSION: Rectal intussusception in patients with evacuatory dysfunction is more advanced morphologically than that seen in asymptomatic controls; it is predominantly full thickness in patients and mucosal in controls. However, caution is required when selecting patients for intervention based solely on radiological findings. PMID- 15898122 TI - Surgery for non-localized bronchiectasis. AB - BACKGROUND: Resection for localized bronchiectasis is a well established therapy. However, there is little information on the role of surgery in non-localized bronchiectasis. METHODS: Between January 1992 and April 2001, 55 patients without cystic fibrosis underwent resection. Forty-eight patients (mean age 45 (range 23 74) years; 32 women) were available for long-term follow-up. Twenty-five patients underwent resection for localized disease (group 1) and 23 had bronchiectasis in at least two different lobes (group 2). RESULTS: Thirty-one of the 48 patients were treated by Video Assisted Thoracoscopic Surgery (VATS) resection. There was no 30-day mortality. Mean duration of hospital stay was 10.9 (range 6-31) days in group 1 and 11.1 (range 5-19) days in group 2. Three of 25 patients in group 1 required reoperation. Only minor complications occurred in group 2 (three patients). Mean follow-up for both groups was 37 (range 6-97) months. Twenty three of 25 patients in group 1 and 16 of 23 in group 2 reported satisfaction at 6 months after the operation. Recurrent infection was noted in three patients in each group. Haemoptysis recurred in only one patient in group 2. CONCLUSION: The surgical treatment of selected patients with non-localized bronchiectasis was safe and most patients were satisfied with the outcome. PMID- 15898123 TI - The relationship between serum prolactin levels and disease activity in patients with Behcet's disease. AB - The aim of this study was to determine the serum levels of prolactin in patients with Behcet's disease and to evaluate its correlation with disease activity. Serum prolactin levels were measured by a chemiluminescence method in 32 patients with Behcet's disease and compared with 20 age-and sex-matched healthy controls. The patients with Behcet's disease were subdivided into two groups according to disease activity: active (18 patients; 13 men and five women, average age 34.0 +/ 6.5 (28-48) years), and inactive (14 patients; 10 men and four women, average age 32.7 +/- 3.1 (22-49 years). Patients with active Behcet's disease had higher serum prolactin levels than the inactive and control groups. Prolactin levels in patients with active Behcet's disease differed significantly from the healthy control subjects (p < 0.05) only, but not the inactive group. Four patients out of 32 (12.5%) Behcet's disease patients showed mild hyperprolactinemia. All four of these cases were from the active Behcet's disease group. Prolactin levels were correlated with ESR (p < 0.05) and CRP (p < 0.05) levels in the active BD group, but not in the inactive BD and control groups. Our results suggest a possible role for this immunoregulatory hormone in the disease expression and pathogenesis of Behcet's disease. PMID- 15898124 TI - Serum iron and copper status and oxidative stress in severe and mild preeclampsia. AB - Our aim was to investigate parameters of iron and copper status and oxidative stress and antioxidant function in women with healthy pregnancy, mild and severe preeclampsia with a view to exploring the possible contribution of these parameters to the aetiology. Thirty healthy, 30 mild preeclamptic and 30 severe preeclamptic pregnant women were included. Serum and placental lipid peroxides, and serum vitamin E and total carotene levels were measured by colorimetric assay. Cholesterol, copper, iron, total iron binding capacity (TIBC), ceruloplasmin and transferrin concentrations were measured by commercially available procedures. Data were analysed statistically using one-way analysis of variance and Pearson correlation test. Logistic regression procedures were used to calculate odds ratios. Lipid peroxides in serum and placental tissue, and iron, copper and ceruloplasmin levels in serum were significantly increased, and transferrin, TIBC, vitamin E/total cholesterol and total carotene/total cholesterol ratios in serum were significantly decreased especially in women with severe preeclampsia. Significant correlations were detected between serum iron and lipid peroxides in serum and placental tissue and between serum iron and vitamin E/total cholesterol in severe preeclamptic pregnancy. Furthermore, there were significant correlations between serum malondialdehyde and ceruloplasmin and vitamin E/total cholesterol in women with severe preeclampsia, and changes in serum and placental lipid peroxides and serum iron concentrations were significantly associated with preeclampsia. In conclusion, ischaemic placental tissue may be a primary source of potentially toxic iron in preeclampsia and the released iron species may contribute to the aetiology and would exacerbate lipid peroxidation and endothelial cell injury, which may be abated by antioxidant supplementation. PMID- 15898125 TI - Comparative study of antioxidant enzymes in tissues surrounding implant in rabbits. AB - There is a possible role of reactive oxygen species (SROS) in the complication of implants although there is presently little information. The aim of this study was to investigate the alterations in lipid peroxidation (LP) and antioxidant enzyme activities in tissues surrounding implants in rabbits. Thirty New Zealand albino male rabbits were used. They were randomly divided into five groups. The first group (I) was used as control. Groups II, III, IV and V were implanted with stainless steel, ceramic, titanium and polyethylene, respectively. One month after the administration of implant, the tissues surrounding the implant were carefully removed for antioxidant enzyme analysis. Glucose-6-phosphate dehydrogenase (G6PD), glutathione reductase (GR), superoxide dismutases (SOD), glutathione peroxidase (GPx), catalase (CAT) in tissues surrounding the implants in the groups II, III and IV were significantly (p<0.05-p<0.001) lower than in the control group although glutathione S-transferase (GST) activities and LP values were increased. CAT activity and LP level did not decrease in group V. In conclusion, these data demonstrate that there is an increase in lipid peroxidation in the tissues surrounding ceramic and titanium implants of animals whereas there is a decrease in antioxidant enzymes. Oxidative stress plays a very important role in the complications of ceramic and titanium implants. The polyethylene implant seems to be the best of the four implant materials tested. PMID- 15898126 TI - Detection of mRNA transcripts of truncated opiate precursor (POMC) in human cartilage. AB - In the present study, we have investigated the presence of pro-opiomelanocortin C terminal fragment derived-peptides in human articular cartilage and cultured chondrocytes. beta-Lipotropin and beta-endorphin were monitored in different cell cultures and biopsies using different techniques. Biopsies were taken from patients undergoing total knee arthroplasty due to osteoarthritis. Both fresh tissue sections and chondrocytes cultured in monolayer were used in the study. Immunohistochemistry, immunocytochemistry, reverse transcriptase-polymerase chain reaction and qualitative Western blots were carried out. The results of the reverse transcriptase-polymerase chain reaction showed transcription of a truncated-form of mRNA for pro-opiomelanocortin in native cartilage and cultured chondrocytes. There was no detection of endogenous production of beta-lipotropin or beta-endorphin in human articular chondrocytes, either in situ or in vitro. Whether pro-opiomelanocortin-derived peptides of non-cartilaginous origin are present in articular cartilage itself still remains unclear. PMID- 15898127 TI - Contribution of the pudendal nerve to sensation of the distal rectum. AB - BACKGROUND: Anal and rectal sensory mechanisms and pudendal nerve function are important in the control of faecal continence. The contribution of the pudendal nerve to sensation of the distal rectum was investigated. METHODS: Heat thresholds in the anal canal, distal and mid rectum were measured using a specially designed thermoprobe. Rectal sensory threshold volumes were measured using the balloon distension method. Needle electrodes were inserted into the external anal sphincter. Pudendal nerve block was performed through a perineal approach, and completeness assessed by loss of electromyographic activity. Heat and rectal volume thresholds were measured again following unilateral and bilateral pudendal nerve block. RESULTS: The technique was successful in four of six volunteers. Bilateral pudendal nerve block produced complete anaesthesia to heat in the anal canal (P = 0.029), but had no effect on heat thresholds in the distal or mid rectum. Rectal sensory threshold volumes were also unaffected by pudendal nerve anaesthesia. CONCLUSION: Anal canal sensation is subserved by the pudendal nerve, but this nerve is not essential to nociceptive sensory mechanisms in the distal or mid rectum. The transition between visceral control mechanisms in the lower rectum and somatic mechanisms in the anal canal may have functional importance in the initiation of defaecation and the maintenance of continence. PMID- 15898128 TI - Characterization of sialidase from bloodstream forms of Trypanosoma vivax. AB - Sialidase (EC: 3.2.1.18) from Trypanosoma vivax (Agari Strain) was isolated from bloodstream forms of the parasite and purified to apparent electrophoretic homogeneity. The enzyme was purified 77-fold with a yield of 32% and co-eluted as a 66-kDa protein from a Sephadex G 110 column. The T. vivax sialidase was optimally active at 37 degrees C with an activation energy (E(a)) of 26.2 kJ mole(-1). The pH activity profile was broad with optimal activity at 6.5. The enzyme was activated by dithiothreitol and strongly inhibited by para-hydroxy mercuricbenzoate thus implicating a sulfhydryl group as a possible active site residue of the enzyme. Theenzyme hydrolysed Neu5Ac2,3lac and fetuin. It was inactive towards Neu5Ac2,6lac, colomic acid and the gangliosides GM1, and GDI. Initial velocity studies, for the determination of kinetic constants with fetuin as substrate gave a V(max) of 142.86 micromol h(-1) mg(-1) and a K(M) of 0.45 mM. The K(M) and V(max) with Neu5Ac-2,3lac were 0.17 mM and 840 micromole h(-1) mg( 1) respectively. The T. vivax sialidase was inhibited competitively by both 2,3 dideoxy neuraminic acid (Neu5Ac2,3en) and para-hydroxy oxamic acid. When ghost RBCs were used as substrates, the enzyme desialylated the RBCs from camel, goat, and zebu bull. The RBCs from dog, mouse and ndama bull were resistant to hydrolysis. PMID- 15898129 TI - Randomized double-blind trial of laparoscopic Nissen fundoplication versus anterior partial fundoplication. AB - BACKGROUND: This double-blind, randomized study compared outcomes of laparoscopic Nissen total fundoplication and anterior partial fundoplication carried out by a single surgeon in a private practice. METHODS: All patients with proven gastro oesophageal reflux disease, regardless of motility, presenting for laparoscopic antireflux surgery were randomized to either Nissen total or anterior partial fundoplication. Primary outcome measures were dysphagia and abolition of reflux. Secondary outcome measures were Visick scores, bloating, patient satisfaction and reoperation rate. RESULTS: Complete follow-up was available for 161 (98.8 per cent) of 163 patients (84 Nissen, 79 anterior). There were no differences in mean heartburn scores between groups. Recurrent reflux was observed in ten patients after anterior fundoplication, but none after the Nissen procedure. Dysphagia scores for both liquids and solids were lower after anterior fundoplication. Four patients had persistent troublesome dysphagia after Nissen fundoplication compared with none after anterior fundoplication. There were no differences between groups in postoperative bloating. The overall reoperation rate at 2 years was 7 per cent, all achieved laparoscopically. CONCLUSION: Nissen fundoplication cured reflux in all patients up to 2 years, but 5 per cent required revisional surgery. Recurrent reflux was more common after anterior fundoplication, but dysphagia was rare. Patient satisfaction was excellent in both groups. Revisional laparoscopic surgery was safe and usually successful. PMID- 15898130 TI - Randomized clinical trial of laparoscopic Nissen fundoplication compared with proton-pump inhibitors for treatment of chronic gastro-oesophageal reflux. AB - BACKGROUND: Both laparoscopic Nissen fundoplication (LNF) and proton-pump inhibitor (PPI) therapy are established in the treatment of gastro-oesophageal reflux disease (GORD). The aim of this study was to compare these two treatments in a randomized clinical trial. METHODS: Between July 1997 and August 2001, 340 patients with a history of GORD for at least 6 months were investigated by endoscopy, 24-h pH monitoring and manometry. Of these, 217 were randomized, 109 to LNF and 108 to PPI therapy. The two groups were well matched for age, sex, weight and severity of reflux. Twenty-four-hour pH monitoring and manometry were performed 3 months after treatment, and quality of life was assessed in both groups using the Psychological General Well-being Index and the Gastrointestinal Symptom Rating Scale at 3 and 12 months after treatment. RESULTS: At 3 months there was an improvement in lower oesophageal sphincter pressure from 6.3 to 17.2 mmHg in the LNF group but no change in the PPI group (8.1 and 7.9 mmHg before and after treatment respectively) (P < 0.001). The mean DeMeester acid exposure score improved from 42.7 to 8.6 (P < 0.001) in the LNF group and from 36.9 to 17.7 in the PPI group (P < 0.001). The mean gastrointestinal symptom and general well being scores improved from 31.7 and 95.4 respectively before treatment to 37.0 and 106.2 at 12 months after LNF, compared with changes from 34.3 and 98.5 to 35.0 and 100.4 respectively in the PPI group. The differences in both of these scores were significant between the two groups at 12 months (P = 0.003). CONCLUSION: LNF leads to significantly less acid exposure of the lower oesophagus at 3 months and significantly greater improvements in both gastrointestinal and general well-being after 12 months compared with PPI treatment. PMID- 15898132 TI - "Double duty" heart pacemakers prove cost-effective. PMID- 15898131 TI - The relationship between time since registration and measured incidence rates in the General Practice Research Database. AB - BACKGROUND: The General Practice Research Database (GPRD) is widely used to study incidence rates. This study examines whether incidence rates are overestimated during the first year after registration, how long one needs to wait to obtain accurate incidence rates, and whether the time period of overestimation differs among disease types. METHODS: We measured incidence rates of nine acute, eight chronic, and eight neoplastic outcomes in 3-month intervals through month 36 after enrollment in GPRD. The incidence rates in months 13-36 were used to estimate baseline incidence rates for each diagnosis. RESULTS: For patients registering with practices that were already UTS, incidence rates were highest in the first 3 months after registration. In eight of nine acute diagnoses, the incidence rate was within 20% of baseline by months 4-6; and in seven of eight cancers, the incidence rate was within 20% of baseline by months 7-9. For chronic conditions, the incidence rate in months 10-12 differed from baseline by more than 20% for five of the eight outcomes, respectively. For patients registering prior to UTS, incidence rates during the first quarter were within 20% of baseline for all acute and cancer diagnoses and six of eight chronic diagnoses. CONCLUSIONS: Reported incidence rates are highest in the first 3 months after registration with an UTS practice and decline to baseline over the first year, more quickly for acute conditions than chronic conditions. For patients who registered prior to UTS, incidence rates are near the baseline level at the start of follow-up. PMID- 15898133 TI - Emergency treatment for cardiac arrest falls short of guidelines. PMID- 15898134 TI - Ask the doctors. I had a stroke while I was traveling. My regular doctor at home asked me what kind of stroke it was. I hadn't realized that there was more than one. What did he mean? PMID- 15898135 TI - Ask the doctors. A doctor told a relative of mine that she needed a stress test. But the test won't involve walking on a treadmill. Instead, she will get a drug to raise her heart rate as if she were exercising. Can you reassure me that this is safe for a person who's not in the best of health? PMID- 15898136 TI - A tax to prevent the epidemic of lung cancer. PMID- 15898137 TI - No place for conflict of interest. PMID- 15898138 TI - Brazil leads from the front on AIDS. PMID- 15898139 TI - Chechen doctors battle to treat the republic's ills. PMID- 15898140 TI - US pharmacies vow to withhold emergency contraception. PMID- 15898141 TI - Wilfred Gordon Bigelow. PMID- 15898142 TI - Skeletal and renal manifestations of Candida tropicalis within a diabetic patient (2005:2b). PMID- 15898143 TI - Compaction, compression and drug release properties of diclofenac sodium and ibuprofen pellets comprising xanthan gum as a sustained release agent. AB - Compaction and compression of xanthan gum pellets were evaluated and drug release from tablets made of pellets was characterised. Two types of pellets were prepared by extrusion-spheronisation. Formulations included xanthan gum, at 16% (w/w), diclofenac sodium or ibuprofen, at 10% (w/w), among other excipients. An amount of 500 mg of pellets fraction 1000-1400 microm were compacted in a single punch press at maximum punch pressure of 125 MPa using flat-faced punches (diameter of 1.00 cm). Physical properties of pellets and tablets were analysed. Laser profilometry analysis and scanning electron microscopy of the upper surface and the surface of fracture of tablets revealed that particles remained as coherent individual units after compression process. Pellets were flatted in the same direction of the applied stress evidencing a lost of the original curvature of the spherical unit. Pellets showed close compressibility degrees (49.9% for pellets comprising diclofenac sodium and 48.5% for pellets comprising ibuprofen). Xanthan gum pellets comprising diclofenac sodium experienced a reduction of 65.5% of their original sphericity while those comprising ibuprofen lost 49.6% of the original porosity. Permanent deformation and densification were the relevant mechanisms of compression. Fragmentation was regarded as non-existent. The release of the model drug from both type of tablets revealed different behaviours. Tablets made of pellets comprising ibuprofen released the model drug in a bimodal fashion and the release behaviour was characterised as Case II transport mechanism (release exponent of 0.93). On the other hand, the release behaviour of diclofenac sodium from tablets made of pellets was anomalous (release exponent of 0.70). For the latter case, drug diffusion and erosion were competing mechanisms of drug release. PMID- 15898144 TI - If only in a dream. PMID- 15898145 TI - Five ways to tackle toxicity (or the ant, the spider, and the bee). 6th International Workshop on Adverse Drug Reactions and Lipodystrophy in HIV. October 25-28, 2004, Washington, DC. PMID- 15898146 TI - Multiple HCV genotype infection speeds HIV progression. PMID- 15898147 TI - Aspirin and women--age matters. Aspirin can help healthy women over age 56 prevent both strokes and heart attacks. For younger women, the hazards of aspirin outweigh the benefits. PMID- 15898148 TI - A better way to prevent blood clots? A drug derived from cobra venom offers an alternative to warfarin, although liver problems temper its benefits. PMID- 15898149 TI - Taking care of yourself is risky business. Understanding risk, and how it is presented, can help you make better decisions about your health. PMID- 15898150 TI - Music still playing for the LDL limbo. PMID- 15898151 TI - Trio of kidney disease signs takes toll on heart. PMID- 15898152 TI - Ask the doctor. In a new study I heard about, people who had leaky mitral valves but no symptoms benefited from an operation to fix the valve. Many of the people in the study had mitral valve prolapse, as I do, but my doctor says I don't have a heart murmur. Would surgery help me? PMID- 15898154 TI - Arthroplasty of the hip. A new operation. PMID- 15898153 TI - Rupture of the posterior urethra with a review of twelve cases. PMID- 15898155 TI - Sympathomimetic drugs in orthostatic hypotension. PMID- 15898156 TI - Reclamation of the rheumatoid hand. PMID- 15898157 TI - Specific antibodies in heart-disease. PMID- 15898158 TI - Measurement of urinary protein. PMID- 15898159 TI - The design and testing of an ultraviolet irradiated box for hand dressings. PMID- 15898160 TI - Tolbutamide in cirrhosis of the liver. PMID- 15898161 TI - New splint for mallet finger. PMID- 15898162 TI - Psychiatry in general hospitals. Manchester's integrated scheme. PMID- 15898163 TI - Psychiatric treatment in general wards. PMID- 15898164 TI - Relationship of doctors and politicians. The Minister's Winchester Address. PMID- 15898165 TI - Brugada syndrome: report of the second consensus conference. AB - Since its introduction as a clinical entity in 1992, the Brugada syndrome has progressed from being a rare disease to one that is second only to automobile accidents as a cause of death among young adults in some countries. Electrocardiographically characterized by a distinct ST-segment elevation in the right precordial leads, the syndrome is associated with a high risk for sudden cardiac death in young and otherwise healthy adults, and less frequently in infants and children. Patients with a spontaneously appearing Brugada ECG have a high risk for sudden arrhythmic death secondary to ventricular tachycardia/fibrillation. The ECG manifestations of Brugada syndrome are often dynamic or concealed and may be unmasked or modulated by sodium channel blockers, a febrile state, vagotonic agents, alpha-adrenergic agonists, beta-adrenergic blockers, tricyclic or tetracyclic antidepressants, a combination of glucose and insulin, hypo- and hyperkalemia, hypercalcemia, and alcohol and cocaine toxicity. In recent years, an exponential rise in the number of reported cases and a striking proliferation of articles defining the clinical, genetic, cellular, ionic, and molecular aspects of the disease have occurred. The report of the first consensus conference, published in 2002, focused on diagnostic criteria. The present report, which emanated from the second consensus conference held in September 2003, elaborates further on the diagnostic criteria and examines risk stratification schemes and device and pharmacological approaches to therapy on the basis of the available clinical and basic science data. PMID- 15898166 TI - First psychosis prodrome: rehabilitation and recovery. AB - This paper explores some of the literature and the personal experience of the author to establish the key feature of the prodrome in first psychosis schizophrenia, the loss of functioning the individual experiences. The emerging literature on the subject is seen to lack consensus, and be focused on the predictive value of prodrome symptoms, not on potential rehabilitation and recovery issues. Because prodrome symptoms are disabling, experienced during the transition from adolescent to adult, and of considerable duration, they are likely to have a profound effect on rehabilitation and recovery in treatment. PMID- 15898167 TI - The next steps forward in reducing the impact of preterm birth. Proceedings of the 2nd International Preterm Labour Congress. 2-4 September 2004, Montreux, Switzerland. PMID- 15898168 TI - Special issue on Richard M. Zaner. PMID- 15898169 TI - Health continuum and data exchange in Belgium and in the Netherlands. Proceedings of Medical Informatics Congress (MIC 2004) and 5th Belgian e-Health Conference. PMID- 15898170 TI - Are noroviruses emerging? PMID- 15898171 TI - Persistent emergence of dengue. PMID- 15898172 TI - More dengue, more questions. PMID- 15898173 TI - Dengue virus type 3, Cuba, 2000-2002. PMID- 15898174 TI - Nosocomial dengue by mucocutaneous transmission. PMID- 15898175 TI - Barriers to Creutzfeldt-Jakob disease autopsies, California. PMID- 15898176 TI - Q fever wildlife reservoir. PMID- 15898177 TI - Vaccine-derived poliovirus, Thailand, 2003. PMID- 15898178 TI - Toscana virus and acute meningitis, France. PMID- 15898179 TI - Helicobacter pylori, Republic of Georgia. PMID- 15898180 TI - Botulism and preserved green olives. PMID- 15898182 TI - Public Health Service policies on research misconduct. Final rule. AB - This final rule removes 42 CFR part 50, subpart A, "Responsibilities of Awardee and Applicant Institutions for Dealing With and Reporting Possible Misconduct in Science," and replaces it with a new, more comprehensive part 93, "Public Health Service Policies on Research Misconduct." The proposed part 93 was published for public comment on April 16, 2004. The final rule reflects both substantive and non-substantive amendments in response to public comments and to correct errors and improve clarity, but the general approach of the NPRM is retained. The purpose of the final rule is to implement legislative and policy changes applicable to research misconduct that occurred over the last several years, including the common Federal policies and procedures on research misconduct issued by the Office of Science and Technology Policy on December 6, 2000. PMID- 15898183 TI - Stroke and bleeding as an endpoint in atrial fibrillation studies. AB - Atrial fibrillation is the most common arrhythmia and is an independent risk factor for stroke and dementia. Stroke endpoints in atrial fibrillation (AF) studies have been defined differently in the past. Consensus is needed in future AF trials and therefore endpoints should be redefined with accuracy. Depending on type of intervention and patient characteristics in the trial, atrial cardioembolic strokes or all strokes should be used. Atrial cardioembolism is very frequent in AF patients and a combined clinical and imaging approach may help to distinguish cardioembolism from other strokes. Accurate identification of type of stroke is mandatory for the optimal evaluation of interventions in AF patients. To that end, cerebrovascular cardiovascular imaging is instrumental. Intracerebral bleeding is an adverse effect of antithrombotic therapy. Therefore, it is an appropriate primary endpoint in trials evaluating antithrombotic strategies. In studies evaluating combined clinical strategies against stroke, i.e., antiarrhythmic and antithrombotic, as well as strategies focused on maintaining vascular integrity, all major cardiovascular and cerebrovascular events should be included in the endpoint. At present, surrogate endpoints for stroke are not being used. Their role remains to be investigated. PMID- 15898184 TI - In memory of Professor Patricia S. Goldman-Rakic. PMID- 15898185 TI - Genetics, ethics and ethnicity. PMID- 15898186 TI - Transition from protozoa to metazoa: an experimental approach. PMID- 15898187 TI - Individuality in early eukaryotes and the consequences for matazoan development. PMID- 15898188 TI - Evolution of animal lectins. PMID- 15898189 TI - Molecular phylogeny of Eumetazoa: genes in sponges (Porifera) give evidence for monophyly of animals. PMID- 15898190 TI - Homeobox genes in the freshwater sponge Ephydatia fluviatilis. PMID- 15898191 TI - Homeobox-containing genes in freshwater sponges: characterization, expression, and phylogeny. PMID- 15898192 TI - Early evolution of the Metazoa: an inference from the elongation factor-1alpha. PMID- 15898195 TI - A professional nurses union. PMID- 15898196 TI - Leadership should unite, not divide. PMID- 15898197 TI - Licensure makes all the difference. PMID- 15898198 TI - Labor class for nurses highly practical. PMID- 15898199 TI - Keep summertime fun: summer safety tips nurses need to know. PMID- 15898202 TI - Governor's Mental Health Services Planning Council to coordinate new freedom commission recommendation: Executive Order 04-10. PMID- 15898203 TI - Creation of governor's commission on healthcare cost containment: Executive Order 04-14. PMID- 15898204 TI - Cast your bread upon the waters? The debate over enforced charity. PMID- 15898205 TI - Health plans, employers join forces to promote e-prescribing. PMID- 15898206 TI - When doctors quit, pay is not the main reason. PMID- 15898207 TI - Insurers give substance abuse new identity: it's a disease. PMID- 15898208 TI - Time to decipher legislation's DNA. PMID- 15898209 TI - Starting Medicare Advantage plan brings special set of problems. PMID- 15898210 TI - Can managed care programs rein in Medicare's runaway budgets? PMID- 15898211 TI - Clinical and economic effects of coronary restenosis after percutaneous coronary intervention in a managed care population. AB - PURPOSE: The epidemiology of coronary restenosis after percutaneous coronary intervention (PCI) has been documented extensively in clinical trials, but no data exist on the clinical and economic burden of restenosis in a managed care population. DESIGN: Retrospective cohort with a nationally representative managed care claims database (IHCIS, Waltham, Mass.) representing 2.8 million members. METHODOLOGY: Patients undergoing initial PCI between 1/1/00 and 12/31/00 (N=3,258) were identified and followed to 1 year. Clinical events, resource use, and costs between 1 month and 1 year after the initial PCI were identified. The clinical restenosis rate was estimated by multiplying the observed repeat revascularization rate by 0.85, based on previously published studies. All costs are reported from a managed care perspective in Year 2000 dollars. PRINCIPAL FINDINGS: Overall, 14.7 percent of patients required 1 or more repeat revascularization procedures between 1 month and 1 year after initial PCI, which implies an estimated clinical restenosis rate of 12.5 percent. Mean 1-year costs were nearly 6-fold higher among patients with and without repeat revascularization (dollars 31,954 +/- dollars 31,857 vs. dollars 5,474 +/- dollars 12,006, P<.001). After adjusting for baseline imbalances, the independent incremental cost for each patient with repeat revascularization was dollars 24,955 (95 percent confidence interval, dollars 23,401-dollars 26,510). Annual follow-up costs attributable to restenosis were dollars 3,118 per initial PCI recipient (i.e., dollars 24,955 x 12.5 percent). CONCLUSION: Clinical restenosis occurred in approximately 12.5 percent of real-world managed care PCI patients and increased health care costs by an average of dollars 3,118 per patient. These findings have important implications for the cost-effectiveness of new treatments that substantially reduce restenosis. PMID- 15898212 TI - Specialty pharmacy meets niche company expertise. PMID- 15898213 TI - Diabetes costs rise, but so does adherence. PMID- 15898214 TI - Don't forget productivity gains when rating health programs. PMID- 15898215 TI - Medicare Modernization Act to increase public sector spending nearly 50 percent. PMID- 15898216 TI - Advancing quality in a consumer-directed world: "May you live in interesting times". PMID- 15898217 TI - The role of accreditation in an era of market-driven accountability. AB - Accreditation has been widely used to promote accountability in healthcare. However, with the rise of both purchaser and consumer demand for broader and more detailed information on performance beyond licensure and professional self regulation, especially at the provider level, the role of accreditation is less clear. We hypothesize that for accreditation to be a critical part of a market driven, consumer-focused healthcare system, accrediting bodies must enlarge their scope of assessment with an emphasis on clinical performance of providers, revise and expand their level of reporting and transparency of assessment, and broaden the base of their governance. A new approach to accreditation could enhance accountability by (1) building on an existing framework and data-collection structure that are proven elements of quality assurance in multiple healthcare sectors; (2) expanding existing involvement of both public and private entities in the process; (3) building on existing linkages to professional and regulatory bodies; (4) providing greater flexibility, compared with regulation, in responding to change; and (5) having a defined source of funding. By these means, accrediting bodies will both improve accountability and successfully drive quality improvement. PMID- 15898218 TI - Point-of-Service reminders for prescribing cardiovascular medications. AB - OBJECTIVES: To provide physicians with evidence-based recommendations for care at the point of service, using an automated system, and to evaluate its effectiveness in promoting prescriptions to prevent cardiovascular events. STUDY DESIGN: Randomized controlled trial. METHODS: Patients at risk for cardiovascular events who might benefit from angiotensin-converting enzyme inhibitors or 3 hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) were identified from electronic data in a managed care organization and randomly assigned into 2 groups. Physicians seeing outpatients in the intervention group were faxed a sheet with pertinent patient data, including a recommendation to prescribe the indicated medication. In the control group, the data sheet did not include the recommendation. Dispensed prescriptions were compared between groups. RESULTS: More than 4000 visits were observed for each medication type. Angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers were dispensed in 7.1% of visits in the intervention group versus 5.7% in the control group (P = .048) following the first patient-physician encounter. No significant difference was observed for statins (intervention, 8.1% vs control, 7.7%). Data for all patient-physician encounters and both medications were combined in logistic regression analysis. The odds ratio was 1.19 for a dispensed prescription in the intervention group and 1.54 for 2 or more visits versus 1 visit. CONCLUSIONS: An automated system that provides pertinent data and tailored recommendations for care at the point of service modestly increased prescription dispensing rates. Targeting patient-provider encounters to change provider behavior is challenging; however, even small effects can produce clinically important results over time. PMID- 15898219 TI - Determinants of compliance with statin therapy and low-density lipoprotein cholesterol goal attainment in a managed care population. AB - OBJECTIVE: To identify determinants of medication compliance and low-density lipoprotein cholesterol goal attainment. STUDY DESIGN: This retrospective analysis used claims data from a large, national, employment-based independent practice association database. Subjects were identified based on the existence of a filled prescription for statin therapy between April 1, 1999, and June 30, 2001. Subjects had to be 18 years or older, continuously enrolled in the health plan for 2 years, and new users of statin therapy. METHODS: Multivariate logistic regression models were used to identify predictors of compliance and goal attainment in high-risk subjects. RESULTS: As the mean copayment for statins increased, there was a decrease in the likelihood of compliance. Of the subjects with laboratory results, 50.7% attained their low-density lipoprotein cholesterol goal level established by National Cholesterol Education Program Adult Treatment Panel III guidelines. Older individuals and men were more likely to reach their low-density lipoprotein cholesterol target goal, as were individuals who were compliant with their statin therapy. CONCLUSIONS: Compliance with statin therapy in the managed care setting remains poor. Of particular concern is the lower level of compliance among women and younger high-risk patients, along with patients who have fewer outpatient visits associated with hyperlipidemia and lower incidences of cholesterol testing. PMID- 15898220 TI - Economic analysis of targeting chemotherapy using a 21-gene RT-PCR assay in lymph node-negative, estrogen-receptor-positive, early-stage breast cancer. AB - OBJECTIVE: To appraise the economics of a recurrence score (RS), based on an assay that predicts distant recurrence-free survival in lymph-node-negative (LN ), estrogen-receptor-positive (ER+) patients with early-stage breast cancer receiving tamoxifen. STUDY DESIGN: Cost-utility analyses using a decision analytic model. METHODS: Using a Markov model, we forecast overall survival, costs, and cost effectiveness of using the RS in patients classified as having low or high risk of distant recurrence based on National Comprehensive Cancer Network (NCCN) clinical guidelines. Data from a large multicenter clinical trial (NSABP B-14) were analyzed to derive risk classification based on guideline criteria and RS assignments. Efficacy of adjuvant chemotherapy (CT) on distant recurrence-free survival (DRFS) was based on published meta-analyses of CT trials. The analysis took a societal perspective, considering survival, quality of life, and relevant costs. RESULTS: Fifty-three patients (8%) were classified as having low risk of distant recurrence by NCCN guidelines and the RS reclassified 15 of these patients (28%) to an intermediate/high-risk group. The remaining 615 patients (92%) were classified at high risk of distant recurrence by NCCN guidelines and the RS reclassified 300 of these patients (49%) to a low risk group. Among a hypothetical cohort of 100 patients, RS is predicted on average to increase quality-adjusted survival by 8.6 years and reduce overall costs by $202 828. RS was cost saving in more than two-thirds of probabilistic simulations, with cost effectiveness most influenced by the propensity to administer CT based on RS results, and by the proportion of patients at low risk as defined by NCCN guidelines. CONCLUSIONS: The RS predicts more accurately than current guidelines recurrence risk in LN-, ER+ patients with early-stage breast cancer. If applied appropriately, the assay is predicted to increase quality adjusted survival and save costs. PMID- 15898221 TI - Validity of the HEDIS criteria to identify children with persistent asthma and sustained high utilization. AB - BACKGROUND: The most widely used performance measure for asthma, the Health Plan Employer Data and Information Set (HEDIS), has been criticized because the delay between classification (year 1) and assessment of medication dispensing (year 2) may produce a "misalignment" and weaken the validity of the measure. OBJECTIVE: To examine whether a previously observed association between the HEDIS performance measure and asthma-related emergency department visits is robust when the period between the classification and outcome assessment is evaluated during a 2-year period as defined. METHODS: Children (N = 2766) aged 3 to 15 years enrolled in 1 of 3 managed care organizations with at least 1 asthma diagnosis listed for a hospitalization, an emergency department visit, or an ambulatory encounter and at least 2 consecutive years of data for analysis from July 1996 through June 1999 were identified. RESULTS: Children did not consistently meet the HEDIS criteria for persistent asthma, and 24% to 28% of children did not requalify in year 2 of observation. Multivariate regression models showed that a protective relationship between controller medication dispensing and asthma related emergency department visits was no longer seen among children meeting the HEDIS criteria for persistent asthma when the total period of observation is extended to 2 years (odds ratio, 0.7; 95% confidence interval, 0.4-1.2). CONCLUSIONS: Our results suggest that the variable nature of asthma may affect how the HEDIS performance measure should be used for assessing quality of care. The period between identification of the target population and performance assessment should be closely related in time. PMID- 15898222 TI - A statewide collaboration to monitor diabetes quality improvement among Wisconsin health plans. AB - OBJECTIVE: The Wisconsin Collaborative Diabetes Quality Improvement Project was initiated in 1999 by the Wisconsin Department of Health and Family Services, Diabetes Prevention and Control Program to monitor quality of diabetes care among the state's health plans. STUDY DESIGN: Prospective observational. METHODS: Annual invitations were mailed to all Wisconsin managed care plans to participate in the project. Collaborators submitted Comprehensive Diabetes Care Health Plan Employer Data and Information Set (HEDIS) measures, as well as other selected HEDIS measures. Data were summarized and reported anonymously back to the collaborators at quarterly forums and in annual reports. RESULTS: Five of the 6 Comprehensive Diabetes Care HEDIS measures have improved significantly in Wisconsin since 1999. Despite this improvement, measure variation across health plans remains high. Collaborators have continued to share resources and best practices at quarterly forums and through statewide initiatives. CONCLUSIONS: This project is an example of an ongoing statewide collaborative quality improvement effort among otherwise competing health plans. Collaboration at regular forums, sharing of HEDIS data to assess quality of diabetes care in health plans, and sharing of resources and best practices may have contributed to improvements in the quality of diabetes care in Wisconsin. PMID- 15898223 TI - Clinicians working in healthcare systems have an important role in early detection and management of overweight in their patients. PMID- 15898224 TI - Reid warning for service: record funding is set to end. PMID- 15898225 TI - Labour's manifesto hints at the shape of things to come. PMID- 15898226 TI - On the 'safety' paradox. PMID- 15898227 TI - Crunch time for managers [interview by Daniel Martin]. PMID- 15898228 TI - Barometer. PCTs March 2005. PMID- 15898229 TI - Mental health. Beyond the fringe. AB - Black and minority ethnic patients are more likely to be diagnosed with schizophrenia and detained, more likely to receive high doses of medication and less likely to have access to talking therapies. Input from voluntary organisations and monitoring staff attitudes are crucial factors in tackling the problem. PMID- 15898230 TI - On mental health. PMID- 15898231 TI - Clinical management. Break even. AB - The orthopaedic project was formed to help the specialty hit December's six-month maximum wait target. s The NOP haemployed an approach of targeted e most challenged trusts. seThe project nds in a team of experts that helps problems and find solutions that will work locally. PMID- 15898232 TI - Buildings & facilities. Project planning. Constructive criticism. PMID- 15898233 TI - Work-related stress. Pressure group. PMID- 15898234 TI - Training and development. Get with the programme. PMID- 15898235 TI - Is it O.K to be pudgy? PMID- 15898236 TI - Cuts will put HIV research back 10 years, scientists warn. PMID- 15898237 TI - GP data transfer could take 18 years. PMID- 15898238 TI - How managers administered a painful kick in the teeth. PMID- 15898239 TI - On post-election priorities. PMID- 15898240 TI - Charge of the fight brigade. PMID- 15898241 TI - The HSJ debate. The public and politicians can learn to value NHS managers. PMID- 15898242 TI - Barometer. Mental health March 2005. PMID- 15898243 TI - A year in Bristol. Two steps forward... AB - UBHT was ahead of the 98 per cent A&E target, but shut a ward to save money and fell far short when winter pressures bit. Its two-star rating is under threat. The trust has had some success in clawing back its forecast deficit, down from pounds sterling 3.5m in December to pounds sterling 1m. UBHT is also undergoing a major structural reorganisation that will put clinicians in the driving seat. PMID- 15898244 TI - On clinical management. PMID- 15898245 TI - Regional focus: Wales. Keep a good thing going. PMID- 15898246 TI - Equality. Crossing the barriers. PMID- 15898247 TI - On her own terms. AB - Considered eccentric in both manner and dress, 'doctress' Mary Seacole became a heroine of the war in the Crimea - but one who has only recently returned to prominence. Unlike her contemporary Florence Nightingale, she concentrated on hands-on care, rather than trying to change the system. PMID- 15898248 TI - Origin of the species. AB - This article explains how antibiotic resistance occurs and how it has grown since the introduction of antibiotics in the 1940s. It charts the rise of M RSA infections and describes their growing threat. PMID- 15898249 TI - Class struggle. PMID- 15898250 TI - Taking your rightful place. PMID- 15898251 TI - No one to blame. PMID- 15898252 TI - Nurses' and doctors' knowledge of pain after surgery. AB - AIM: To analyse the links between levels of acute pain management knowledge, perceptions of clinical skills and the acute pain management education history of doctors and nurses working in orthopaedics and general surgery in an acute hospital. METHOD: Questionnaires were sent to 101 doctors and nurses. Eighty two were returned. Questions were derived from an abridged version of McCaffery and Ferrell's (2002) Nurses' Knowledge and Attitudes Survey Regarding Pain, acute pain literature and the trust's clinical standards and protocols. RESULTS: The results identified the most useful sources of acute pain management education and who accessed these; barriers to good acute pain management, other than a lack of education; differences between doctors and nurses in standards of education, levels of perceived competence and knowledge in assessment, pharmacology and analgesic delivery systems. CONCLUSION: Staff and patient misconceptions, inadequate sources of learning and professional traditions continue to affect the quality of acute pain management. However, progress has been made in the trust to overcome these factors through collaborative practice, review of education programmes and specific practice development initiatives. PMID- 15898253 TI - Supporting adolescents through behaviour change. AB - This article, the fifth of eight in the health promotion series, aims to inform practitioners about potentially harmful health and lifestyle-related behaviours in adolescents, and to provide some strategies for supporting young people to make changes. PMID- 15898254 TI - Management of complications in early pregnancy. AB - This article examines complications in early pregnancy and outlines their management in an early pregnancy unit. It includes a description of early fetal development. The use of ultrasonography as a diagnostic tool and the psychological impact of early pregnancy loss are discussed. PMID- 15898255 TI - Hip replacement. PMID- 15898256 TI - Ladder of success. PMID- 15898257 TI - Bad report card. Adverse safety cases on the rise in hospitals, study says. PMID- 15898258 TI - The fix is in for Medicaid. With dollars 10 billion in Medicaid spending reductions approved by Congress, the next battle is over forming the commission. AB - It could have been worse. But then it could have been better. That was how healthcare providers assessed the Medicaid funding approved by Congress last week as part of the fiscal 2006 budget. Cuts in projected spending totaled dollars 10 billion over four years. "We fought against there being any cuts, but we think these are somewhat palatable," says the American Hospital Association's Tom Nickels, left. PMID- 15898259 TI - Beating the clock. Congress asks 10 largest systems for billing records. PMID- 15898260 TI - Out of reach. Cover the Uninsured Week highlights coverage crisis. PMID- 15898261 TI - Upping the ante. The CMS' transfer policy changes anger hospitals. PMID- 15898262 TI - Getting ready for '75% rule'. Providers adjust to limits on rehab reimbursement. PMID- 15898263 TI - Strong message. Fletcher Allen official receives two-year sentence. PMID- 15898264 TI - Making amends. Bills could signal victory for Fla. docs, hospitals. PMID- 15898265 TI - Drug samples bring scrutiny. PMID- 15898266 TI - Specialties have smaller profits. PMID- 15898267 TI - Forward march. IBM strengthens healthcare presence through UPMC, Healthlink deals. PMID- 15898268 TI - Knowing when to quit. O'Leary's hedging opens doors for others on patient safety. PMID- 15898270 TI - By the numbers. 25 busiest community hospital emergency departments ranked by number of emergency room visits, 2003. PMID- 15898269 TI - Quality as gospel. They might be devoted to different approaches and processes, but healthcare executives are taking on quality improvement with almost religious zeal. PMID- 15898271 TI - Marburg haemorrhagic fever, Angola--update. PMID- 15898272 TI - Global status of immunization safety: a report based on the WHO/UNICEF Joint Reporting Form, 2003 update. PMID- 15898274 TI - 2005 MSMS physician data survey on practice characteristics. PMID- 15898273 TI - Producing medical records in litigation. PMID- 15898275 TI - Michigan's physicians benefit from favorable liability climate. PMID- 15898276 TI - "Michigan steps up" to improve health and lifestyles--and so does MSMS. PMID- 15898277 TI - Congressman Schwarz talks about needed systemic changes. Interview by Jennifer V. Higgins. PMID- 15898278 TI - Change coming for Varicella reporting in Michigan. PMID- 15898280 TI - One family of medicine. PMID- 15898279 TI - Obesity--a major health issue. PMID- 15898281 TI - In contrast with PAR-2 mediated nociception and hyperalgesia, melanosome transfer does not involve N-methyl-D-aspartate receptors. PMID- 15898282 TI - Mechanism-based treatment of acne vulgaris: the value of combination therapy. AB - Acne vulgaris has multiple pathogenic mechanisms that act in concert to produce disease. Effective therapy addresses more than one pathogenic factor to speed resolution of disease. Typically, retinoids are used to inhibit comedo formation and an antibacterial is used to suppress Propionibacterium acnes (P. acnes). Using combinations of agents can enhance efficacy, but increase side effects. Therefore, the tolerability of a topical retinoid is important. Strategies to enhance retinoid tolerability include the use of new retinoids such as adapalene, new delivery systems such as the tretinoin microsphere delivery system, or alternative dosing schedules such as short contact or every other day (QOD). PMID- 15898283 TI - A chart review of patients with early stage mycosis fungoides treated with psoralen plus UVA (PUVA). AB - PUVA has become a common form of treatment for early stage mycosis fungoides (MF). The purpose of this retrospective study was to review the clinical data of 51 MF patients (96% stage IA or IB) treated with PUVA at the Mt. Sinai MF clinic over the past 20 years. We analyzed the efficacy, safety, and remission duration in patients who were treated with a modified PUVA regimen. Forty-four of 51 patients (86%) achieved complete clinical clearing for all stages after initial PUVA therapy. The mean duration of remission with maintenance treatment was more than 27 months (range: 3 weeks to 130 months). The mean duration of disease from start of first PUVA therapy for all patients was 4.8 years (range: 0.7 to 130 months). PUVA for patients with early-stage MF is a safe and effective therapeutic modality with prolonged disease-free remissions, however, PUVA alone was not adequate for more advanced disease. PMID- 15898284 TI - Therapeutic decision making in the therapy of actinic keratoses. AB - Actinic keratoses (AKs) represent the second most common reason to visit a dermatologist in the United States and their therapy has become a major portion of most dermatologists' practice. An ever-increasing array of therapeutic options exist for the therapy of actinic keratoses, offering physicians and patients a greater number of choices than ever before. Patient expectations and needs seem to be changing at the same time, thus effecting therapeutic decision-making. While destructive therapies with resultant wounds, time for wound healing, and possible hypopigmentation or scarring were acceptable in the past, many patients from the baby-boom generation are now developing AKs and have little tolerance for any time for wound healing or any cosmetic changes. This paper will raise some fundamental questions regarding AKs and their management. PMID- 15898285 TI - The use of terbinafine in the treatment of onychomycosis in adults and special populations: a review of the evidence. AB - Terbinafine is an allylamine with fungicidal activity, first approved for the treatment of onychomycosis in the United Kingdom in the early 1990s, and in the US in 1996. Terbinafine is the most frequently prescribed oral antifungal agent in the US and Canada for onychomycosis. Its efficacy and safety in dermatophyte toenail onychomycosis in adults has been established in many studies. In fact, 18 randomized controlled trials have shown terbinafine to be highly effective, with a meta-average for mycological cure of 76% +/- 3% (mean +/- standard error). In large surveillance studies, terbinafine exhibited excellent safety profiles consistent with results obtained in pivotal studies. Additionally, terbinafine has been reported to be superior to both itraconazole and fluconazole in comparative studies in the treatment of dermatophyte toenail onychomycosis. Recent studies have reported terbinafine to be more cost effective than griseofulvin, fluconazole, or itraconazole. Terbinafine has also been used to treat onychomycosis effectively and safely in special patient populations, such as children, the elderly, immunocompromised patients, diabetics, and those with Down syndrome. Terbinafine should therefore be considered for the management of onychomycosis in adults based on its effectiveness, broad spectrum, fungicidal nature, established safety profile, and very low occurrence of drug interactions. Furthermore, the data support the use of terbinafine to treat dermatophyte onychomycosis in children and the elderly. PMID- 15898286 TI - The effect of lidocaine and adrenaline on the viability of injected adipose tissue--an experimental study in nude mice. AB - Adipose tissue injection as a free graft for the correction of soft tissue defects is a widespread procedure in plastic surgery. The main problem in achieving long-term soft tissue augmentation is partial absorption of the injected fat, and hence the need for over-correction and re-injections. Lidocaine, used for local anesthesia, has been suspected as an inhibitor of growth of adipocytes in culture and slowing down glucose transport and lipolysis in adipocytes. In addition, reduced blood supply by local application of epinephrine was accused of having a negative effect on the graft. The purpose of this study was to examine the effect of local anesthesia, administered to the fat donor site, on the take of the injected fat. Human adipose tissue, obtained by suction-assisted lipectomy, was injected subcutaneously into the scalp of nude mice. Local anesthesia of the fat donor site consisted of a solution with 600 mg of lidocaine (0.06%) and epinephrine 1:1000000. In a control group, normal saline with no local anesthesia or epinephrine was administered to the donor site. One cc of fat was injected after centrifugation into each animal scalp. There were 10 animals in each group. The animals were sacrificed 15 weeks after the procedure. Graft weight and volume were measured, and histologic evaluation was performed. No significant differences were demonstrated between the groups in regard with the grafts' weight and volume and the histologic parameters investigated. In conclusion, local anesthesia solution, consisting of lidocaine and epinephrine, does not alter the take of fat grafts, and has no influence on the adipocytes viability. PMID- 15898287 TI - The potential of interleukin 12 inhibition in the treatment of psoriasis. AB - Interleukin 12 (IL-12) is an important cytokine produced by a variety of immune effector cells that leads to a type 1 helper T cell (Th1) response. IL-12 also directs T cells to the skin via induction of cutaneous lymphocyte antigen (CLA) expression. In this article we report the current understanding of the immunobiology of IL-12, reviewing its structure, receptor, and function. We also discuss the role of IL-12 in the pathogenesis of psoriasis. Some effective conventional psoriasis treatments alter IL-12 levels. Importantly, specific antibodies directed against IL-12 may prove useful against psoriasis but may also act by targeting IL-23 in addition to IL-12. PMID- 15898288 TI - Rosacea, light, and phototherapy. AB - The long-established notion that rosacea is worsened by light is of particular concern in the phototherapy of diseases such as psoriasis, eczema, or vitiligo, which often can be coexistent with rosacea. A literature search was conducted and much evidence was found to challenge this belief that light adversely affects rosacea. In fact, more patients actually improved with sunlight in a more recent published survey. Several other studies have also shown that rosacea patients were similar to control subjects in sun exposure, solar skin damage, and sun sensitivity. Additionally, all clinical trials to date have failed to find a difference between rosacea patients and control subjects when challenged with ultraviolet light. Thus, phototherapy with rosacea may be safer than is commonly believed. PMID- 15898289 TI - Clobetasol propionate 0.05% lotion in the treatment of moderate to severe atopic dermatitis: a randomized evaluation versus clobetasol propionate emollient cream. AB - Atopic dermatitis (AD) is a chronic inflammatory and pruritic skin disorder marked by alternating periods of relapse and remission. This multicenter, randomized, active- and vehicle-controlled, investigator-blinded study compared the efficacy and safety of clobetasol propionate lotion to clobetasol propionate cream formulation and lotion vehicle in the treatment of moderate to severe AD. A total of 229 subjects applied treatment twice-daily for 2 weeks. Clobetasol propionate lotion was significantly more effective than its lotion vehicle at 2 weeks and comparable to the cream formulation. Clinical success after a 2-week, treatment-free follow-up period was greater in the clobetasol propionate lotion group than in the cream group. Clobetasol propionate lotion is effective, safe, well tolerated and offers a better remission profile in the treatment of moderate to severe AD as compared to clobetasol propionate emollient cream. PMID- 15898290 TI - A randomized controlled clinical trial assessing the effect of betamethasone valerate 0.12% foam on the short-term treatment of stasis dermatitis. AB - BACKGROUND: There are no published studies examining either the effectiveness of topical steroids in the treatment of stasis dermatitis or indicating what steroid strength or duration of treatment is optimal to treat this common condition. OBJECTIVE: To investigate the efficacy of twice-daily application of the topical steroid betamethasone valerate 0.12% foam for the treatment of stasis dermatitis. DESIGN: 42-day randomized, double-blinded, vehicle-controlled, pilot study. SETTINGS: Outpatient dermatology clinic at a university-affiliated clinic. SUBJECTS: 19 subjects, mean age of 73, with mild to moderate bilateral stasis dermatitis. INTERVENTION: Twice-daily application of betamethasone valerate 0.12% foam versus vehicle foam to bilateral randomly assigned lower legs for 28 days with follow-up to day 42. MAIN OUTCOME MEASURES: The primary clinical endpoints were the mean change in erythema, scale, swelling, petechiae, post-inflammatory hyperpigmentation, and self-reported pruritus, assessed on a 5-point Likert scale (0 = clear, 1 = almost clear, 2 = mild, 3 = moderate, 4 = severe). Secondary endpoints were changes in health related quality of life (HRQL) using the EuroQol 5D (EQ-5D) utility score and visual analog scale (VAS) and the Dermatology Life Quality Index (DLQI). RESULTS: Although there was no overall difference between the foam and vehicle-treated leg at days 14 and 28, the steroid-treated leg, but not the vehicle-treated leg, showed statistical improvement over baseline. Improvement in the steroid-treated leg was statistically better than vehicle at days 14 and 28 in terms of erythema (P < .05) and petechiae (P < .05). Improvement in VAS was notable at days 14 (7.1%), 28 (9.7%), and 42 (9.6%) (P < .001). Similarly, there was a statistically significant improvement in the DLQI compared to baseline on visit days 14 (188.9%) and 28 (126.1%) (P < .001). CONCLUSIONS: This study suggests that betamethasone valerate 0.12% foam is an effective and well-tolerated short-term treatment of stasis dermatitis, but that higher potency steroids may be needed to achieve better efficacy. Furthermore, these results are the first to suggest that the application of effective topical anti-inflammatory therapy can lead to improvement in HRQL. PMID- 15898291 TI - Evaluation of subcision as a treatment for cutaneous striae. AB - BACKGROUND: Treatment options for cutaneous striae are very limited. OBJECTIVE: To determine if subcision is an effective treatment for cutaneous striae and compare a combination with 0.1% tretinoin cream against a single treatment. METHODS: 14 patients were evaluated, each with 3 white striae. One stria received treatment with subcision, one with tretinoin cream in a 0.1% concentration, and one with subcision plus tretinoin. The length of follow-up was 3 months. Two blind investigators evaluated improvement by comparison of the initial and final digital photographs. RESULTS: Only 7 patients completed 3 months follow-up. A decrease of width and clinical improvement was observed with the 3 treatments. However, some striae showed no change at all and there was no statistically significant difference between treatments. Three patients presented necrosis in striae treated with subcision. CONCLUSIONS: There are no studies in the medical literature about subcision for the treatment of cutaneous striae. This is a preliminary study and considering the undesirable effects of necrosis in a high percentage of striae treated with subcision, the subjective way of evaluation, and the small study group, we can not recommend subcision as a treatment for cutaneous striae. Large studies are necessary to corroborate or to discard our findings. PMID- 15898292 TI - Basal cell carcinoma of the areola in a man. AB - Basal cell carcinomas (BCCs) that arise in sun-protected sites are quite uncommon. We report a case of BCC of the areola, which is extremely rare. Mohs micrographic surgery was used for the treatment. The etiologic factors of BCCs in sun-protected areas are discussed, and previous studies regarding the treatment modalities are reviewed. PMID- 15898293 TI - Merkel cell carcinoma in a patient with autoimmune hepatitis. AB - Merkel cell carcinoma (MCC) has been shown to have a higher incidence in many etiologically distinct immunosuppressed populations. We report a case of aggressive MCC diagnosed in a man with autoimmune hepatitis who was treated with immunosuppressive therapy for more than 30 years. PMID- 15898294 TI - The successful use of etanercept in combination therapy for treatment of acrodermatitis continua of hallopeau. AB - Acrodermatitis continua of Hallopeau (ACH) is a rare form of pustular psoriasis which poses a challenge to treat and causes considerable pain and suffering for those afflicted. Tumor necrosis factor-alpha (TNF-alpha) is a pro-inflammatory cytokine involved in the pathogenesis of ACH and other forms of psoriasis. Inhibition of TNF-alpha has been shown to provide benefit in such inflammatory conditions as rheumatoid arthritis, psoriatic arthritis, and, most recently, plaque psoriasis. In this report, we present the case of a 65-year-old man with a 9-year history of recalcitrant ACH who demonstrated significant and sustained clinical improvement when etanercept, a competitive inhibitor of TNF-alpha, was added to his treatment regimen of acitretin and topical corticosteroids over a 12 week period. PMID- 15898295 TI - Furuncular myiasis secondary to Dermatobia hominis. AB - Dermatobia homininis is the most common cause of furuncular myiasis in Central and South America. It is diagnosed based on a history of travel to an endemic region and the characteristic cutaneous lesion. We present a 54-year-old patient who presented with both a travel history and cutaneous findings of furuncular myiasis. PMID- 15898296 TI - Isotretinoin in acne vulgaris: a prospective analysis of 160 cases from Kuwait. AB - A total of 160 patients (59 male and 101 female) ages varying from 13 to 28 years (mean age 20 years) with moderate to severe acne were treated with isotretinoin in the doses ranging from 0.5 mg/kg/day to 1 mg/kg/day. The drug was given for a period ranging from 6 to 28 weeks. The patients were followed up regularly for a period of 12 months after stoppage of isotretinoin for any evidence of relapse. In the event of a recurrence greater than mild acne after 8 weeks of stoppage of isotretinoin therapy, the patients were given another course of the drug. Patients were considered to be non-relapsing if they had no evidence of recurrence after 12 months of follow-up. Twenty seven patients were excluded from the study. Of the remaining 133 patients (51 male and 82 female) only 117 patients (36 male and 81 female) could follow up for at least 12 months after stopping therapy. Of the 133 patients, a total of 127 patients (95.5%) achieved complete or partial clearance. Forty two percent (total 49 patients: 20 male and 29 female) experienced relapse after stopping therapy. Of these, 21 (42.85%) were given a second course of the drug. None of the patients developed a rise in lipids levels significant enough to warrant stoppage of the drug. PMID- 15898297 TI - Clofazimine as elective treatment for granulomatous cheilitis. AB - Cheilitis granulomatosa (CG) is a rare, idiopathic inflammatory disorder that usually affects young adults and clinically is characterized by diffuse, non tender, soft to firm swelling of one or both lips. A variant of granulomatous cheilitis is Melkersson-Rosenthal syndrome when associated with facial paralysis and furrowed tongue. Several treatments have been used with variable success. We report 3 cases of GC treated with oral clofazimine 100 to 200 mg daily for 3 to 6 months obtaining regression of lesions in all treated cases. Hyperpigmentation and elevation of liver enzymes were observed as side effects. PMID- 15898298 TI - Herpes acquisition and transmission. AB - Present dogma is based on herpesviruses (HSV-1 and HSV-2) residing only in the dorsal root ganglion and laying dormant unless reactivated. Polymerase chain reaction has improved specificity and sensitivity so that tenets based on previous methodologies can be questioned. On point, negative serology, such as with anti-glycoprotein G protein antibodies for HSV-1 and HSV-2, does not rule out the presence of herpes. Indeed, exposure to the virus is probably universal, with individuals displaying varied immunological responses to the herpesviruses. There is a bimodal temporal distribution of herpetic reactivation, explained by the existence of the virus in the epidermis as well as in the dorsal root ganglion. Additionally, herpes may share a symbiotic relationship with humans, in which the body actually retains the virus within numerous body organs for its possible anti-cancer properties. PMID- 15898299 TI - The ABCDEs of melanoma: an evolving concept. PMID- 15898300 TI - Poliomyelitis outbreak spreads across Yemen; case confirmed in Indonesia. PMID- 15898301 TI - Marburg haemorrhagic fever, Angola. PMID- 15898302 TI - Epidemic-prone disease surveillance and response after the tsunami in Aceh Province, Indonesia. PMID- 15898303 TI - Comparing child outcomes of physical punishment and alternative disciplinary tactics: a meta-analysis. AB - This meta-analysis investigates differences between the effect sizes of physical punishment and alternative disciplinary tactics for child outcomes in 26 qualifying studies. Analyzing differences in effect sizes reduces systematic biases and emphasizes direct comparisons between the disciplinary tactics that parents have to select among. The results indicated that effect sizes significantly favored conditional spanking over 10 of 13 alternative disciplinary tactics for reducing child noncompliance or antisocial behavior. Customary physical punishment yielded effect sizes equal to alternative tactics, except for one large study favoring physical punishment. Only overly severe or predominant use of physical punishment compared unfavorably with alternative disciplinary tactics. The discussion highlights the need for better discriminations between effective and counterproductive use of disciplinary punishment in general. PMID- 15898304 TI - Interpersonal relationships and sex differences in the development of conduct problems. AB - This article investigates the role of interpersonal relationships in shaping sex differences in the manifestation, etiology, and developmental course of conduct problems and their treatment needs. The review examines whether: (1) Girls' conduct problems are more likely than boys' to manifest as a function of disrupted relationships with caretakers and peers; (2) For girls more than for boys, the outcomes of conduct problems in adolescence and adulthood, and related treatment needs, are more likely to be a consequence of the quality of interpersonal relationships with others, particularly opposite-sex peers and partners. Evidence reviewed suggests that boys and girls share many similarities in their expression of conduct problems, but that a relational perspective does unify important differences. There is fair evidence that girls with conduct problems are more likely to come to the attention of authorities because of chaotic, unstable family relationships, and to express antisocial behavior in the context of close relationships; there is stronger evidence that the course and outcomes of conduct problems in females versus males pertain to interpersonal relationship impairments. Those sex differences map onto specific differences in treatment needs. Further empirical testing of the proposed relational model is indicated. PMID- 15898305 TI - Test anxiety: a cross-cultural perspective. AB - The present paper examines test anxiety from a cross-cultural perspective with specific reference to the Indian and American cultures. The construct of test anxiety has been examined in many cultures all over the world. In this review, the importance of understanding and incorporating contextual factors in cross cultural research is emphasized. Moreover, some of the methodological issues related to investigating culture-behavior relationship are discussed. Specifically, the derived-etic approach for conducting cross-cultural research is espoused. Then, research findings from western, cross-cultural, and Indian studies on test anxiety are reviewed. Consistent with the individualistic orientation of the western society, much of the research in the western world has adopted a de-contextualized approach. Inasmuch as many of the cross-cultural and Indian studies on test anxiety have their roots in western research, they have ignored the cultural context as well. To address this void, contextual variables relevant to test anxiety in the Indian setting are examined and hypotheses regarding the nature of test anxiety in Indian children are proposed. Finally, a research agenda is presented to examine these hypotheses using a derived-etic approach. PMID- 15898308 TI - Revving up revenue. PMID- 15898309 TI - Skin lesions and crawling sensations: disease or delusion? PMID- 15898310 TI - ICD-10 codes: help or headache. PMID- 15898311 TI - Food allergies: avoidance is the answer. PMID- 15898312 TI - Recurrent vulvovaginitis candidiasis; an overview of traditional and alternative therapies. AB - Vulvovaginitis is a common inflammatory condition that can significantly disrupt a woman's life. The term vulvovaginitis actually encompasses a variety of inflammatory lower gential tract disorders that may be secondary to infection, irritation, allergy or systemic disease (Table 1). This article focuses on candida-caused vulvovaginitis, a condition known as vulvovaginitis candidiasis. Approximately 75% of U.S. women expreience vulvovaginitis candidiasis during thier reproductive years. Between 40% and 50% of these women have recurrent episodes, and 5% to 8% experience chronic candida infections. Approximately 3 million women have recurrent candidial infections. The prevalence of vulvovaginitis candiasis is expected to rise due to the growing number of non-C albicans species (which are immune to most antifungal medcationa) and as a result of more widespread antifungal resistance. PMID- 15898313 TI - Common functional bowel problems. What do homeopathy, Chinese medicine and nutrition have to offer? PMID- 15898314 TI - Massage therapy: integration with traditional medicine. PMID- 15898315 TI - Chaste tree for premenstrual syndrome. An evolving therapy in the United States. PMID- 15898316 TI - Untangling the confusion. Alzheimer's management today. PMID- 15898317 TI - Controversial, confident and committed: how a close-knit group of believers launched the NP profession. Interview by Jolynn Tumolo. PMID- 15898318 TI - Intimate partner violence: a culturally sensitive approach. PMID- 15898319 TI - Flu-shot politics. PMID- 15898320 TI - Angela Bull's letter in our February issue has attracted a number of responses... PMID- 15898321 TI - Angela Bull's letter in our February issue has attracted a number of responses... PMID- 15898322 TI - Angela Bull's letter in our February issue has attracted a number of responses... PMID- 15898323 TI - Angela Bull's letter in our February issue has attracted a number of responses... PMID- 15898324 TI - Work it. Claim it. PMID- 15898325 TI - Do aged care employers value their nurses? PMID- 15898326 TI - Bullying. PMID- 15898327 TI - Brand new day? PMID- 15898329 TI - Case studies in wage recovery. PMID- 15898328 TI - Discrimination against enrolled nurses. PMID- 15898330 TI - Employees' basic rights under threat by Howard Government. PMID- 15898331 TI - The blind man sees... PMID- 15898332 TI - [The rabbit IOP and pupil values after application of aminoacid L-lysine and antiglaucomatic Timoptol mixture]. AB - OBJECTIVE: In this article authors focused to influence the physiologic IOP values after application of the aminoacid 10% L-lysine.2HCl.2H2O in 0.5% Timoptol eye drops mixture in experiments. METHODS: 0.5% Timoptol, followed by 10% L lysine.2HCl.2H2O and mixture of 10% L-lysine.2HCl.2H2O with 0.5% Timoptol was instilled weekly into the left conjunctival sac of 5 adult rabbits (females of the New Zealand White species). The IOP and pupilar diameter were measured in 5, 15, 30, 60, 120, 180 and 240 min. after instillation. The right eye was used as control. RESULTS: In experiments authors observed that: a) the aminoacid 10% L lysine.2HCl.2H2O is without influence on the physiologic value of the IOP; b) 0.5% Timoptol compared with control eye nonsignificantly decreased the rabbit IOP; c) after instillation of 10% L-lysine.2HCl.2H2O and 0.5% Timoptol mixture the IOP showed decrease of the physiologic IOP in two periods. The initial non significant period of the IOP decrease was measured after application up to the 60th min. The identical values were also in the control eye. However, the major decrease of the IOP compared with control eye was observed during the second period starting from 60th min. up to the end of experiments (to the 4th hour) after instillation of this mixture. The mean value of this significant decrease was 3.3 torrs of the physiologic IOP value showing regular and linear effectivity in the experimental eye. In the control eye the second period was without decrease of the IOP. CONCLUSIONS: The results demonstrate that the Timolol alone decreased the physiologic IOP no significantly, the aminoacid L-lysine.2HCl.2H2O is without influence on the physiologic value of the IOP. In experiments we confirmed that in a mixture of aminoacid L-lysine.2HCl.2H2O with antiglaucomatic 0.5% Timoptol a new substance was created, a new "bio-antiglaucomatic", responsible for more significant decrease of the physiologic IOP. PMID- 15898333 TI - [New incision versus corneal flap uncover: comparison of two techniques of repeated surgery after primary LASIK in myopia]. AB - An advantage of the LASIK method is the possibility of repeated surgery in case of residual refractive error after the primary surgery. In the second surgery two techniques are used: the technique of the new incision (secondary LASIK) and the technique of corneal flap uncover. The authors evaluate the results of the residual refractive errors using both techniques of reoperaration. The cohort constituted of 74 reoperated eyes of 61 patients. The average age was 29.3 +/- 7.83 years (18 to 48 years) with the average follow-up period of 6.7 +/- 4.7 months (3 to 12 months). The reoperation was performed with the primary refractive error (before the first surgery) less than -3.0 dioptres (D) in 3 eyes, -3.25 to -6.0 D in 24 eyes, and higher than -6 D in 47 eyes of the cohort. Technique of the new incision was chosen in 21 eyes (28.4%) of the cohort (1st group) and corneal flap uncover in 53 eyes (71.6%) (2nd group). The residual refractive error before the reoperation was -1.3 D +/- 0.94 (standard deviation- SD) combined with -1.17 +/- 0.86 (SD) cylindrical dioptres (Dcyl) in the 1st group, and -1.12 D +/- 1.27 (SD) combined with -0.83 Dcyl +/- 0.64 (SD) in the 2nd one. The primary LASIK surgery, as well as reoperation was in all cases performed by argon-fluoride laser, class IV (Keracor 117 Chiron Technolas). The authors evaluated the final postoperative refractive error, reduction of its spherical and cylindrical component, uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA) the rate of complications during the surgery, and postoperatively as well. The final postoperative refractive error was in the 1st group -0.35 D +/- 0.40 (SD) and -0.56 Dcyl +/- 0.67 (SD), and in the 2nd group 0.17 D +/- 0.36 (SD) and -0.28 Dcyl +/- 0.45 (SD). Reduction of the residual postoperative refractive error after primary LASIK surgery in the spherical part of the error in the new incision technique group was 73.1% (P = 0.01), and in the corneal flap uncover technique 84.8% (P = 0.01). The cylindrical part lowered in the 1st group by 52.1% (P = 0.05), and in the 2nd group by 66% (P = 0.01). Improvement of the uncorrected visual acuity (UCVA) was noticed in the 1st group in 18 eyes (85.7%), and in the 2nd group in 47 eyes (88.7%). The best corrected visual acuity remained in most of the cases unchanged--in the group of the new incision technique in 16 eyes of the cohort, and in the corneal flap uncover technique in 36 eyes (67.9%). Complications during the surgery and after that were noticed in the 1st group in 4 eyes (19%), and in the 2nd group in 12 eyes of the cohort (22.6%). No statistically significant difference in the reduction of the residual refraction error after primary LASIK and complications rate between both mentioned techniques of re-operation were noticed. Both methods may be considered effective and safe. PMID- 15898334 TI - [Bilateral Terrien's degeneration treated by corneoscleral graft transplantation]. AB - Authors operated on a patient with bilateral corneal involvement in Terrien's marginal degeneration and spontaneous corneal perforation. The surgery was performed in both eyes at the same time and a free sclerocorneal graft was used. The postoperative cosmetical and functional results as well were very good. It is advisable to follow up the patients with Terrien's marginal dystrophy. The surgery on right time removes the defect of the cornea and accompanying high corneal astigmatism. PMID- 15898335 TI - [Trabeculectomy--long term results]. AB - Goal of this paper was to analyze retrospectively and partially also prospectively the results of trabeculectomies (TE) performed at the Department of Ophthalmology of the University Hospital in Hradec Kralove (Koniggratz), Czech Republic, EU, during the 1990-2000 period using the database program GLAUCOM Access 97. The authors did statistical analysis of 538 trabeculectomies performed as the first surgery for lowering the intraocular pressure (IOP) on the followed eye without peroperative use of antimetabolites or cytostatics with follow up period 1-10 years. The baseline criteria for the evaluation were changes of the IOP, visual acuity, and pharmacological treatment after the operation. The average age at the time of the surgery was 61.28 +/- 14.58 years. Surgeries were performed by 11 surgeons. During the follow-up period the IOP lowering effect of the surgery was demonstrated by lowering the average IOP values from 28.42 +/- 8.98 mm Hg to 16.53 +/- 5.93 mm Hg. In the first half of the follow-up period a slight improvement of the visual acuity was noticed, in the second one the visual acuity decreased. Pre-operative average visual acuity was 0.59 +/- 0.34, and at the end of the follow-up period it was 0.59 +/- 0.33. The surgery in glaucoma patients effectively lowered the average number of prescribed antiglaucomatics after TE from 1.92 +/- 0.83 to 0.89 +/- 0.80 drugs. Our evaluations clearly showed trabeculectomy is even in the presence of existence non-perforating surgeries the basic and relatively easy-to-perform IOP lowering technique with broad variety of indications, and with relatively low number of complications. PMID- 15898336 TI - [Inflammatory pseudotumor of the orbit]. AB - The authors present a patient who was hospitalized in their department due to indurative edema of the left lower eyelid. Computed tomography (CT) of the orbit confirmed the suspicion of tumor affection behind the globe. The general condition of the patient was not altered. Biopsy and following histological examination confirmed the diagnosis of inflammatory pseudotumor of the orbit. The patient was treated systemically with corticosteroids and the condition normalized. PMID- 15898337 TI - [Transmission electronic microscopy of the inner limiting membrane and epiretinal tissue in idiopathic macular hole]. AB - The authors examined samples of epimacular tissue obtained during the surgeries of the idiopathic macular hole in different stages of the disease by means of transmission electronic microscopy. In the early stages of the disease only the inner limiting membrane with isolated cells on the vitreous side was present, in later ones of the disease the connective tissue membrane was attached. In rare cases of the later stages of the disease they found the presence of the retinal pigment epithelium (RPE). They suppose the RPE plays an important role in the regeneration of the defects of the retina. PMID- 15898338 TI - [Computer program for antiglaucomatous surgeries analysis]. AB - Computer analysis of long-term results of the surgical treatment of glaucoma needs suitable program for database data download retrospectively and prospectively as well. We composed a universal database program Glaucoma Access 97 for patients' follow-up in glaucoma surgery centers. Data about 1073 antiglaucomatous filtering surgeries from the period 1990-2000 were downloaded. The program permits to download data about the patients before the surgery, details about the operation procedure, and from the follow-up visits into the database using special software questionnaire sheets. Outcome from the database may be data about long-term IOP compensation, visual acuity changes, and treatment after the surgery. Evaluation of the data allows to follow up the successfulness of the treatment of specified groups of patients and to compare results of surgeons and individual departments as well. PMID- 15898339 TI - [Acanthamoeba keratitis]. PMID- 15898340 TI - [Chairman of the Prague Ophthalmology Clinic 1935-1939 and a long-serving chief physician in Cheb, Dr. Jaroslav Kubik (4 April 1886--4 September 1980)]. PMID- 15898341 TI - Are stimulators of fat oxidation interesting drug discovery targets? PMID- 15898342 TI - A potential role of odorant receptor agonists and antagonists in the treatment of infertility and contraception. AB - In 1992, the identification of odorant receptor expression in mammalian testicular tissue prepared the ground for an ongoing debate about a potential role for these chemoreceptors in significant sperm behaviors, in particular chemotaxis. The identification of hOR17-4, a human testicular odorant receptor that mediates sperm chemotaxis in various bioassays, revealed the first potential key player in this reproductively relevant scenario. Detailed knowledge of the receptor's molecular receptive field, the discovery of a potent receptor antagonist, as well as specific insight into the receptor-linked signaling cascade(s), could establish a basis for pioneering future applications in fertility treatment and/or contraception. PMID- 15898343 TI - Novel therapeutic targets for appetite regulation. AB - Obesity is currently the major cause of premature death in the UK, killing almost 1000 individuals per week, and worldwide, its prevalence is accelerating. Many peptides are synthesized and released from the gastrointestinal tract and, while their roles in the regulation of gastrointestinal function have been known for some time, it is now evident that they also physiologically influence eating behavior. Therefore, manipulation of gastrointestinal hormones provides the prospect of an effective and well-tolerated treatment for obesity. Whereas drugs targeting appetite-signaling neuropeptides in the brain may also affect other aspects of the central nervous system, agents based on gut hormones themselves have the advantage of targeting specific appetite circuits within the brain without producing any unacceptable side effects. PMID- 15898344 TI - SomatoKine: is there a use in treating growth disorders? AB - Growth hormone (GH) has been used to treat GH deficiency since the late 1950s, and recombinant GH has been available since 1985. GH is also approved to treat non-GH-deficient short stature, such as that seen in Turner syndrome, chronic renal insufficiency, Prader-Willi syndrome, children who are small for gestational age, and idiopathic short stature. There has been interest in using recombinant insulin-like growth factor I (IGF-I) to treat short stature, either alone or in combination with its binding protein, IGF binding protein (IGFBP)-3 (SomatoKine). IGF-I increases growth velocity in children with IGF deficiency, either as a result of growth hormone insensitivity syndrome (GHIS) or IGF-I gene deletion. However, there have been adverse events, particularly hypoglycemia, reported with administration of unbound IGF-I. In addition, the serum half-life of unbound IGF-I is shorter when administered to patients with GHIS, who have low serum concentrations of its primary binding protein IGFBP-3 than when administered to healthy individuals or to patients with an IGF-I gene deletion (who have normal levels of IGFBP-3). SomatoKine was developed to prolong the half life and to counteract acute adverse events (particularly hypoglycemia) associated with IGF-I administration. SomatoKine appears to have a longer half life in patients with GHIS than unbound IGF-I and fewer adverse events (including hypoglycemia) have been reported when administered to patients with diabetes. PMID- 15898345 TI - Prolactin receptor antagonists. AB - Efforts have been focused on the development of prolactin receptor (PRLR) antagonists, which, it is hoped, will be useful therapeutics for breast, prostate and possibly other cancers. Several approaches have been undertaken, including the production of pure antagonists and the combination of a PRLR antagonist with other antitumor molecules to increase efficacy. An alternative approach has been the development of a mimic of the natural PRL growth antagonist. This molecule is a combined antagonist/agonist since it inhibits signaling leading to growth, but promotes signaling leading to cell-cycle control, differentiation and apoptosis. PMID- 15898346 TI - Mitotane for adrenocortical carcinoma treatment. AB - Adrenocortical carcinoma (ACC) is a rare neoplasm with poor prognosis. Mitotane is the only adrenal-specific agent available for the treatment of ACC, and although it has been used for several decades, many of its pharmacological properties, as well as its exact mechanism of action, remain to be fully elucidated. It is known that metabolic activation is essential for the adrenolytic activity of mitotane. Most published clinical reports of mitotane use are retrospective analyses of small numbers of patients, however, the collective findings of these studies indicate that mitotane has activity against ACC; in approximately 25% of cases mitotane led to an objective tumor regression and, in the majority of patients, control of hormone excess could be achieved. Side effects occur frequently during mitotane treatment and they mainly affect the gastrointestinal tract and the central nervous system. As not all patients respond to mitotane therapy, it is one of the challenges for the future to define the subset of patients who do respond to mitotane to avoid treatment of patients who are unlikely to respond to this toxic drug. The pharmacological properties of mitotane and its efficacy in the treatment of ACC are analyzed in this review. PMID- 15898347 TI - Treating diabetic retinopathy by tackling growth factor pathways. AB - Diabetic retinopathy (DR) remains a major cause of new cases of blindness onset in adults. The prevalence of diabetic eye disease is strongly related to the duration of diabetes, blood pressure and glycemic control, although a multifactorial pathogenesis is likely to be probable. Despite the effectiveness of current prevention (by tight metabolic and blood pressure control) and treatment (with laser photocoagulation) methods, and with the help of screening programs, diabetic eye disease is still a problem. Recent advances in our understanding of the pathogenesis of microvascular complications and particularly of the role of growth factors (GFs) in retinal changes have allowed significant advances in the medical management of DR. Studies of the biochemical process underlying DR have clearly demonstrated an important role for a number of aberrantly expressed GFs or their second messengers (eg, vascular endothelial growth factor, growth hormone, insulin-like growth factor-1, protein kinase C and pigment epithelium derived factor) possibly acting together in the development of structural changes characterizing early stages of vascular DR. The critical role of GF expression has led to new experimental therapeutic intervention in DR. In fact, timely pharmacological intervention in GF synthesis and activities may arrest the development of early vascular changes. As the effects of GFs become better understood, pharmacological manipulation of GF synthesis and action will be useful in the early stages of vascular change with the potential to prevent diabetes-related visual loss. PMID- 15898348 TI - Pyridoxamine (BioStratum). AB - BioStratum is developing pyridoxamine (Pyridorin), an advanced glycation end product (AGE) inhibitor, for the potential prevention of diabetic nephropathy. By January 2004, phase II trials had been completed. PMID- 15898349 TI - MK-431 (Merck). AB - Merck & Co is developing MK-431, the lead from a series of dipeptidyl peptidase IV inhibitors that enhance endogenous glucagon-like peptide-1 levels, for the potential treatment of type 2 diabetes. Phase III studies were initiated in the second quarter of 2004. PMID- 15898350 TI - Muraglitazar (Bristol-Myers Squibb/Merck). AB - Bristol-Myers Squibb and Merck & Co are co-developing muraglitazar, a dual peroxisome proliferator-activated receptor-alpha/gamma agonist, for the potential treatment of type 2 diabetes and other metabolic disorders. In November 2004, approval was anticipated as early as mid-2005. PMID- 15898351 TI - Strontium ranelate (Fujisawa/Servier). AB - Strontium ranelate (PROTELOS), a divalent strontium salt, has been developed and launched by Servier for the treatment of osteoporosis. Fujisawa Pharmaceutical is developing the drug in Japan; in November 2004, phase II Japanese trials were ongoing. PMID- 15898352 TI - Functionality of succinylated Brazil nut (Bertholletia excelsa HBK) kernel globulin. AB - One of the possible ways to improve the utilisation of defatted Brazil nut kernel flour, a by-product of oil extraction industries, is to improve its functional properties by chemical modification as it possesses very modest functional characteristics. Succinylated Brazil nut kernel globulin at 55.8%, 62.4% and 72.0% level showed a positive effect on functionality. The solubility of acylated globulin was improved above pH 4.0 but was reduced in the pH range of 3.0-4.0. Water absorption (1.96-4.00, 4.12, and 4.21 ml/g protein), oil absorption capacity (1.44-2.72, 2.80 and 2.94 ml/g protein) and apparent viscosity of the succinylated globulin increased with increase in the level of succinylation. The extent of modification also influenced emulsifying capacity, which showed a decrease at pH 3.0, but was increased at pH 5.0,7.0 and 9.0. Highest emulsion activity (approximately 63.0%) was observed at pH 3.0, followed by pH 9.0 and pH 7.0 and, least (about 11.8%) at pH 5.0. Emulsion stability also followed similar behaviour as that of emulsion activity. The improved functional properties of succinylated Brazil nut kernel globulin could be explored in a variety of food formulations such as high protein drinks, soups, bakery and meat products as well as in salad dressings and mayonnaise as an emulsifier. PMID- 15898353 TI - Isolation and partial characterization of mango (Magnifera indica L.) starch: morphological, physicochemical and functional studies. AB - Mango (Mangifera indica) is a fruit that grows in tropical regions. The aim of this work was to isolate the starch from two varieties of mango highly consumed in Mexico ("criollo" and "manila"), and to evaluate its chemical composition, along with some morphological, physicochemical and functional properties. Mango starch had an amylose content of about 13%, the fat content of "criollo" variety starch (0.1-0.12%), was similar to that of commercial corn starch used as control (0.2%); both mango starches had higher ash amount (0.2-0.4%) than corn starch. Mango starches presented a smaller granule size (10 microm) than corn starch (15 microm), along with an A-type X-ray diffraction pattern with slight tendency to a C-type. All values of water retention capacity (WRC) increased with the temperature. When the temperature increased, solubility and swelling values increased and in general, mango starches had higher values than corn starch. Both mango starches had gelatinization temperatures lower than the control, but "criollo" variety starch presented higher enthalpy values than "manila" variety and corn starches. Overall, it was concluded that due to its morphological, physicochemical and functional properties, mango starches could be a feasible starch source with adequate properties, suitable for using in the food industry. PMID- 15898354 TI - Effects of protein-bound polysaccharide isolated from pumpkin on insulin in diabetic rats. AB - To investigate hypoglycemic substances from pumpkin, protein-bound polysaccharide was isolated by activity-guided isolation from water soluble substances of the fruits. The protein-bound polysaccharide from pumpkin fruits (PBPP) was identified to consist mainly of polysaccharide (approximately 41.21%) and protein (approximately 10.13%) by anthrone test and Lowry-Folin test. Different doses of PBPP were evaluated for hypoglycemic activity and the effect on serum insulin levels in alloxan diabetic rats. The results indicated that PBPP can obviously increase the levels of serum insulin, reduce the blood glucose levels and improve tolerance of glucose. The hypoglycemic effect of big dose PBPP group (1000 mg/kg body weight) excelled that of small dose PBPP group (500 mg/kg body weight) and antidiabetic agent group. The results suggest that the hypoglycemic effect of PBPP depends on the dose and PBPP possesses the possibility of being developed from a new antidiabetic agent. PMID- 15898355 TI - Easy preparation of dietary fiber with the high water-holding capacity from food sources. AB - Dietary fibers were prepared as alkali- and acid-insoluble fractions with chemical phosphorylation from Tossa jute (Corchorus olitorius), defatted soybean (Glycine max), and Shiitake (Lentinula edodes). The dietary fiber fractions treated with alkaline solution containing sodium metaphosphate had the lower protein content and higher total dietary fiber content than those of the preparations without phosphorylation. Alkaline extraction followed by phosphorylation led to a 1.5-fold increase in the water holding capacity of dietary fiber compared with no phosphorylation, whereas the binding capacity to bile acids of dietary fiber was almost the same. The alkali- and acid-insoluble extraction with phosphorylation provided an efficient preparation of water insoluble dietary fiber with high-water holding capacity from various food sources. PMID- 15898356 TI - Antimutagenic properties of green tea. AB - In this work biological effects of two common kinds of green tea (Chinese Gunpowder and Japanese Sencha) were analyzed using three independent tests of antimutagenicity: 1) the Ames test with Salmonella typhimurium TA98, 2) cytogenetic analysis of peripheral blood lymphocytes (CAPL), and 3) test with Saccharomyces cerevisiae D7. Tea extracts were allowed to be antimutagenic based on their ability to inhibit the mutagenic effect of standard mutagens. Amounts of (-)catechin and (-)catechin gallate in tea extracts were determined by high performance liquid chromatography on reversed phase (RP-HPLC). Antioxidant capacity was found using total radical-trapping antioxidant parameter (TRAP) method. Extracts from Gunpowder and Sencha exhibited high antimutagenic activity in the Ames test (24.7+/-3.7% and 34.1+/-2, 1% of inhibition without metabolic activation; 74.9+/-1.7% and 62.7+/-4.3% of inhibition with metabolic activation, respectively) as well as in S. cerevisiae D7 test (Gunpowder: 62.7+/-5.7% of Trp convertants inhibition and 52.6+/-5.3% of Ilv revertants inhibition; Sencha: 45.6+/-4.2% of Trp convertants inhibition, 50.0+/-4.8% of Ilv revertants inhibition). In the CAPL method reduced number of abberant cells as well as decreased number of chromosome breaks was observed using both green tea extracts. Antioxidant capacity and antimutagenicity of green tea extracts was higher than activity of tea catechins and flavonoids. PMID- 15898357 TI - Variations of main quality components of tea genetic resources [Camellia sinensis (L.) O. Kuntze] preserved in the China National Germplasm Tea Repository. AB - The variations of the main quality components of tea, tea polyphenols, catechins, amino acids, caffeine, and water extract of 596 accessions Chinese tea genetic resources, preserved in the China National Germplasm Tea Repository (CNGTR), were analyzed. Tea polyphenols content on a dry weight basis varied from 13.6 to 47.8%, averaging 28.4%. The tea polyphenols content increases gradually from northern and eastern provinces to southern provinces, the highest is in Yunnan tea resources. The content of catechins ranged from 81.9 g/kg to 262.7 g/kg, averaging 144.6 g/kg. The genetic resources from Hunan province had the highest catechins content, inconsistent with the tea polyphenols, which were highest in Yunnan tea resources. The amino acids content ranged from 1.1 to 6.5%, with an average of 3.3%. It was much lower in the southern provinces compared with northern and eastern provinces. The average caffeine content was 4.2%, varying from 1.2 to 5.9%. Yunnan province has plentiful high caffeine genetic resources, next is Fujian province. Caffeine content of Chinese and Japanese tea genetic resources were very similar. The average of water extract content was 44.7%, varying from 24.4 to 57.0%. The variation pattern was similar to that of tea polyphenols. Tea genetic resources of transnormal components are valuable and they could be used directly or indirectly for commercial functional components extract, breeding, and production. PMID- 15898358 TI - Location, complications and treatment of mesiodentes--a retrospective study in children. AB - Out of approximately 11 500 children, who had been referred to the Department of Paediatric Dentistry at the Institute for Postgraduate Education,Jonkoping, Sweden, 97 individuals had a total of 123 mesiodentes. Their dental records and radiographs were analysed in order to study the clinical and radiographic features of the mesiodentes, to record the complications caused by them and to evaluate different forms of treatment. In the present study, the predominant location of the mesiodentes was palatally with 39 per cent in an inverted position. Arrest of the permanent incisors eruption, midline diastema and displacement or rotation of the incisors, were the most common complications found. On the contrary, no follicular cysts or resorption of either the mesiodens or the adjacent teeth were found. Fourty-three out of the 97 individuals were radiographically followed and no pathological findings were recorded. This study shows that the risk of complications caused by the mesiodentes seems to be very low and therefore a more restrictive attitude towards their surgical removal can be recommended. PMID- 15898359 TI - Influence of interleukin (IL)-1 composite genotype on clinical variables in non smoking, well-maintained compliant patients with chronic periodontitis. AB - The aim of this retrospective study was to evaluate the influence of interleukin (IL)-1 genotype on the clinical parameters pocket probing depth and tooth loss in periodontally treated and regularly maintained patients. Only data from non smokers were included. Patient mean data on pocket probing depth (PPD) and tooth loss from 53 Caucasian patients treated for generalized chronic periodontitis and maintained for an average of 15.5 years at the university of Kiel were analysed. All patients were genotyped and subjects with at least one copy of the variant allele 2 at positions IL-1A -889 and IL-1B +3954 were classified IL-1 genotype positive. 12 patients (22.6%) were IL-1 genotype positive. IL-1 genotype positive/negative patients were on average 46.3/45.8 years old with a mean of 23.8/23.9 teeth, resp., at baseline (TO). A total of 86 teeth were lost initially, resulting in an average of 21.5/ 22.5 teeth after active treatment (T1). The corresponding values after 13 years of supportive periodontal care (T2) were 20.7/21.6 teeth. Corresponding mean PPD values for IL-1 genotype positive/negative patients decreased from 4.7 mm/5.3 mm (To) to 2.9 mm/ 2.9 mm (T1) and increased slightly to 3.3 mm/3.4 mm at T2. Intergroup differences of PPD and tooth loss were not significant. Regarding pocket probing depth and tooth loss, there were no significant differences related to IL-1 genotype. PMID- 15898360 TI - Tactile stimulation as a complementary treatment of temporomandibular disorders in patients with fibromyalgia syndrome. A pilot study. AB - Pain of long duration is a common suffering in modern man. One such pain condition is fibromyalgia syndrome (FMS). Opinions about what treatment regimen that are to be used in these patients are diverging, and many of the treatments suggested are not, or only poorly, scientifically investigated. The aim of this pilot investigation was to evaluate if FMS patients with signs and symptoms of temporomandibular disorders (TMDs) refractory to conservative TMD treatment would respond positively to tactile stimulation in respect of local and/or general symptoms. Ten female patients fulfilling the inclusion criteria received such treatment once a week during a 10-week period. At the end of treatment, a positive effect on both clinical signs and subjective symptoms of TMD, as well as on general body pain, was registered. Eight out of 10 patients also perceived an improved quality of their sleep. At follow-ups after 3 and 6 months some relapse of both signs and symptoms could be seen, but there was still an improvement compared to the initial degree of local and general complaints. At the 6-months follow-up, half of the patients also reported a lasting improvement of their sleep quality. One hypothetical explanation to the positive treatment effect experienced by the tactile stimulation might be the resulting improvement of the patients' quality of sleep leading to increased serotonin levels. The results of the present pilot study are so encouraging that they warrant an extended, controlled study. PMID- 15898361 TI - Primary teeth in adults--a pilot study. AB - Remaining primary teeth are occasionally found in adults. The cause and the long term survival may vary. The aim of this investigation was to study the occurrence and condition of primary teeth in adults in a county in Sweden. All dentists in general practice were asked to send in radiographs showing remaining primary teeth in patients aged 18 years and more. The following variables were registered: 1) age, 2) gender, 3) year of exposure of the radiograph, 4) location of the primary tooth, 5) impaction of the permanent successor, 6) fillings or caries in the primary tooth, 7) amount of root resorption and 8) infraocclusion of the primary tooth. 35 dentists contributed with radiographs from 65 subjects showing 89 retained primary teeth. The most commonly found tooth was the second primary molar in the mandible, followed by the primary canine in the maxilla. Except for maxillary molars, most of the primary teeth showed a moderate degree of root resorption. No relationship was found between the degree of root resorption and gender, fillings/caries or infraocclusion. For the mandibular primary molars, there was a statistically significant correlation between root resorption and age. The mesial root of the molar in the mandible was more affected by root resorption than the distal, and the degree of root resorption of the mesial and distal roots was found to be closely correlated. Also, the results indicated a symmetrical pattern of root resorption in bilateral cases of remaining mandibular second primary molars. PMID- 15898362 TI - Detection of lingual root resorptions in the intraoral radiographs. An experimental study. AB - An experimental study was undertaken in an attempt 1) to describe the possibilities to detect lingual root resorptions of upper incisors in intraoral radiographs and 2) to determine the amount of reduction that is necessary in order to detect resorptions in the radiograph. The upper right central incisor was extracted from a dry skull. In a standardized way, the root was repeatedly reduced by grinding on the lingual side at an angle of 16 degrees measured from apex to the long axis of the tooth. After each reduction the tooth was replaced in the dry skull and radiographed in a standardized manner. The radiographs were placed in two different charts and at random within each chart. On two different occasions three groups of observers, 5 orthodontists, 5 pedodontists and 5 radiologists were asked to register any detected "root resorptions" on the right first and second incisors. The first chart included radiographs with a reduction of 0-3 mm and the second chart 0-6 mm. The orthodontists and pedodontists were not very consistent in their observations and false positive registrations were very frequent. The radiologists were more accurate in their observations and detected resorptions when the reduction incorporated the apex of the root at a 3 mm level. Thus, intraoral radiographs seems to be a very blunt instrument for detecting lingual root resorptions. PMID- 15898363 TI - Is radius-union an indicator for completed facial growth? AB - The purpose of this study was to determine if facial or dentoalveolar growth takes place after the occurrence of the radiographic handwrist stage R-J (completed fusion of the proximal epiphysis and diaphysis of the radius), which generally defines the completion of facial growth. Handwrist roentgenograms and lateral head films (LHFs) from 56 subjects (31 male and 25 female) aged 14 to 21 years were examined. Eight cephalometric distances (3 sagittal, 4 vertical, and 1 diagonal) and their changes during 2 different examination periods (from 1 year before to 2 years after the occurrence of R-J) were assessed. The investigation revealed that after the occurrence of R-J vertical dentoalveolar growth changes between 0.5 and 1.75 mm took place in the mandible and in the maxilla in 20% and 10% of the subjects, respectively. Comparing the 2 periods the frequency of the measured growth changes before and after R-J was, on average, almost equal. Because chronological age cannot be used for the assessment of facial growth termination, a reliable indicator for the assessment of facial growth termination is of major importance. After the occurrence of the handwrist stage R-J, most subjects revealed growth changes of less than 1 mm. Especially in the field of tooth implantology, it is important to forecast even small vertical dentoalveolar growth changes. The detected amount of growth after the insertion of an implant can cause esthetic and functional problems. PMID- 15898364 TI - An evaluation of the use of digital study models in orthodontic diagnosis and treatment planning. AB - The purpose of the present study was to determine the diagnostic and treatment planning value of digital models when compared with plaster study casts. In addition, the level of orthodontic experience of the examiner was assessed to determine whether this would have an influence on the decision-making process. Thirty randomly selected orthodontic patients from the Department of Orthodontics at the University of Alabama were selected for the study. From the 30 record sets, seven were selected attempting to mirror cases required for presentation to the American Board of Orthodontics. The seven evaluators were divided into two groups on the basis of their level of orthodontic experience. Initially, each evaluator assessed each patient record. Each evaluator was given a standardized questionnaire which recorded the evaluator's diagnosis based on use of the digital study models (T1). Regardless of whether the evaluator requested a review of the plaster study casts, the evaluator was given the plaster study casts. The evaluator then, using the plaster casts, filled out another identical questionnaire (T2). A chi-square test was used to determine any group differences in the frequency of changed diagnostic characteristics, treatment mechanical procedures, or proposed treatment plans after evaluating plaster study models. The statistical significance selected was P = .05 level of significance. The results showed that 12.8% of diagnostic characteristics, 12% of treatment mechanic procedures, and 6% of proposed treatment plans changed after T2. The results of the present study indicate that in the vast majority of situations digital models can be successfully used for orthodontic records. PMID- 15898365 TI - Mandibular asymmetry in adult patients with unilateral degenerative joint disease. AB - The purpose of this study was to evaluate the association between unilateral degenerative joint disease (UDJD) and lower facial asymmetry. Twenty symptomatic patients and 20 asymptomatic volunteers were studied. Posteroanterior radiographs and bilateral mandibular laminographs were used to evaluate lower face and mandibular asymmetry. The two-way analysis of variance test was used to compare the groups. The posteroanterior radiographs did not reveal vertical asymmetry for the patients. The group with the left side affected exhibited deviation of the mandibular dental midline, the menton, and the antigonion toward the affected side. The laminographs of the same group revealed decreased condylar height and increased antigonial notching in the affected side. The results suggest that subjects with UDJD may manifest lower face asymmetry. This can be of clinical value when adult patients exhibiting asymmetry are to be treated. PMID- 15898366 TI - Classification of the skeletal variation in normal occlusion. AB - The aims of this study were to classify normal occlusion samples into specific skeletal types and to analyze the dentoalveolar compensation in a normal occlusion in order to provide the clinically applicable differential diagnostic criteria for an individual malocclusion patient. Lateral cephalograms of 294 normal occlusion samples, who were selected from 15,836 adults through a community dental health survey, were measured. Using a principal component analysis, two factors representing the anteroposterior and vertical skeletal relationships were extracted from 18 skeletal variables. Cluster analysis was then used to classify the skeletal patterns into nine types. Nine types of polygonal charts with a profilogram were created. Discriminant analysis with a stepwise entry of variables was designed to identify several potential variables for skeletal typing, which could be linked with computerized cephalometric analysis for an individual malocclusion patient. Discriminant analysis assigned 87.8% classification accuracy to the predictive model. It was concluded that because the range of a normal occlusion includes quite diverse anteroposterior and vertical skeletal relationships, classifying the skeletal pattern and establishing an individual dentoalveolar treatment objective might facilitate clinical practice. PMID- 15898367 TI - Abnormal variations in the facial soft tissues of adult uremic patients on chronic dialysis. AB - Renal osteodystrophy is a common complication of uremic patients, mainly when they are in long-term regular dialysis treatment. The craniofacial skeleton is one of the peculiar targets of renal osteodystrophy, the more dramatic pattern of which is a form of leontiasis. To assess facial deformities in uremic conditions and to understand the possible relation with hyperparathyroidism, the three dimensional coordinates of 50 soft tissue facial landmarks were obtained by an electromagnetic digitizer in 10 male and 10 female patients (aged 53-81 years) with chronic renal insufficiency and regular dialysis treatment. Similar three dimensional coordinates were also collected for 34 healthy individuals of the same age, ethnicity, and sex. From the landmarks, facial distances, angles, and volumes were calculated according to a geometrical model of the face. Data were compared with those collected in the normal subjects by computing z-scores. Facial volumes of the chronic renal insufficiency patients were significantly larger than those of the control subjects. The increment was particularly evident in the facial middle third (maxilla), leading to a significant inversion of the mandibular-to-maxillary ratio. The lips tended to be more prominent (reduced interlabial angle). Some of these facial modifications (facial and nose height, maxillary dimensions, mandibular shape) were significantly related to the clinical characteristics of the patients (duration of renal insufficiency, duration of dialysis, parathormone (PTH) plasma levels). In regular dialysis treatment, some of the facial modifications are related to hyperparathyroidism, which supports the need for a strict metabolic control also in the early phase of uremia. PMID- 15898368 TI - Effect of facial convexity on antero-posterior lip positions of the most favored Japanese facial profiles. AB - Soft tissue analyses are basic tools in planning orthodontic treatment. However, the perception of a well-balanced Japanese facial profile has not yet been established, particularly with respect to changes in facial convexity. The purpose of this study was to assess and determine the range of antero-posterior (A-P) lip positions of the top three most favored, well-balanced profiles from a series of various facial silhouettes with changing facial convexity. The average profile was constructed from the profiles of 30 Japanese males and females with Class I occlusions. Changes in facial convexity were established by altering the A-P chin position by protruding or retruding the chin in increments of 5 degrees and 10 degrees from the average profile. A series of 11 profiles were developed from each of the four changes in facial convexity. The lips were then protruded or retruded in one-mm increments from the average profile. Forty orthodontists were asked to rate the top three most favored, well-balanced profiles. We found that the raters tended to prefer more retruded lip positions as facial convexity decreased, for both males and females. The raters also tended to prefer slightly more protruded lip positions as the facial convexity increased. Because chin position is inherent in each patient, others may equate a retrusive or protrusive chin with certain personalities, thus, it is important for orthodontists to take into account the balance between the lips and the chin. PMID- 15898369 TI - Bimaxillary dentoalveolar protrusion: traits and orthodontic correction. AB - A group of 48 ethnically diverse patients with bimaxillary protrusion was used to study the pretreatment cephalometric traits of this malocclusion and the effect of orthodontic correction. All patients were treated with four premolar extractions and retraction of the anterior teeth. Pre- and posttreatment lateral cephalograms were evaluated using a series of 18 linear and angular measurements, and the effect of orthodontic correction was determined using paired t-tests. Cephalometric standards were developed for bimaxillary protrusions, which clarify the overall presentation of this malocclusion for clinicians. Patients with bimaxillary protrusion demonstrated increased incisor proclination and protrusion, a vertical facial pattern, increased procumbency of the lips, a decreased nasolabial angle, and thin and elongated upper and lower anterior alveoli. This study also showed that the extraction of four premolars can be extremely successful in reducing the dental and soft tissue procumbency seen in patients with bimaxillary protrusion, thus providing a stronger evidence-based rationale for this treatment modality. PMID- 15898370 TI - Tooth wear in the mixed dentition: a comparative study between children born in the 1950s and the 1990s. AB - The aim of this study was to evaluate the degree of tooth wear in posterior deciduous teeth of 100 subjects in the second phase of mixed dentition who were born in the 1950s (50sG) and 100 subjects born in the 1990s (90sG). The degree of abrasion for each posterior deciduous tooth was scored ranging from 0 to 3. The comparison of the degree of abrasion showed significant differences between the two groups for all examined teeth (upper and lower deciduous canines and first and second primary molars) all of which appeared to be more abraded in the 50sG. The findings of the present study indicate that subjects who were born in the 1950s exhibited a significantly greater degree of tooth wear on the posterior deciduous teeth when compared with contemporary subjects, along with a well recognized lower prevalence of malocclusions. Changes in dietary habits and a diet based on processed foods may be postulated as factors involved with a decrease of dental attrition in contemporary populations. An increase in the prevalence of mouth breathing (tested in the present study) appears to be associated with a decrease in tooth wear in the contemporary population as well. PMID- 15898371 TI - Orthodontic treatment need from eight to 12 years of age in an early treatment oriented public health care system: a prospective study. AB - The aim of this study was to investigate the effects of an early treatment oriented orthodontic program on treatment need at age 12 years in a public health care system. The subjects consisted of one whole age cohort in a municipality in Finland, 87 children in total. All children were examined at ages eight, 10, and 12 years. Treatment need was assessed on casts using a modified Dental Health Component of the Index of Orthodontic Treatment Need and a 10-Grade Scale based on the Treatment Priority Index. Early treatment was started on children having definite treatment need according to both indices. The agreement between indices was good at ages eight and 12 years and moderate at age 10 years. Treatment need changed significantly from eight to 12 years. Of the 29 children with definite treatment need at age eight years, only two had treatment need at age 12 years. Of the 38 children with no treatment need at age eight years, 28 remained in this category and only two children had definite need for treatment at age 12 years. The duration of treatment ranged from one to 61 months, although 59% of the treatments lasted two years or less. We conclude that systematically planned early orthodontic treatment may have contributed to the significant reduction in treatment need from eight to 12 years of age. PMID- 15898372 TI - Simulation study for a finite helical axis analysis of tooth movement. AB - The description of motion using a finite helical axis (FHA) is independent of the chosen coordinate system because the values of the helical axis parameters do not change with coordinate transformations. However, an intuitive understanding of tooth movement expressed by the FHA can be slightly difficult for orthodontists who have never used it. The purpose of this study was to clarify the basic behavior of the FHA to increase our understanding of the FHA during tooth movement. Parameters of the FHA were calculated in two different simulations of canine retraction. In simulation 1, as the tipping angle of the canine was increased from 5 degrees to 30 degrees, the direction vector of the FHA v approached the x-axis (tipping axis), thus increasing the rotation angle about the FHA theta. These results demonstrated that the direction vector of the FHA can indicate the axis of rotation that is most affected in three-dimensional transformation. In simulation 2, bodily tooth movement of the canine was increased from one to five mm, and the shortest distance from the origin to the FHA d increased in a linear manner. In this case, the percent increase in bodily tooth movement could be determined by the d value. This study showed that the use of FHA makes it possible to determine the torques applied to the tooth and the true bodily tooth movement during orthodontic treatment, and this may lead to a better understanding of how to move teeth. PMID- 15898373 TI - Evening primrose oil effects on osteoclasts during tooth movement. AB - This study was conducted to investigate the influence of supplemented evening primrose oil (EPO) on osteoclast expression during experimental tooth movement in rats. Forty-eight 10-week-old male Sprague-Dawley rats were divided into experimental and control groups. Animals in the experiment group were fed a 7.25 g/kg daily dose of EPO orally by gastric intubation for 20 days before orthodontic tooth movement. The animals in the control group received an equivalent volume of distilled water by the same method. On day 21, a 40-g mesial tipping force was applied to the maxillary right first molar of each rat. After loading, six animals in each group were sacrificed on days 0, 3, 7, and 14 with the appliance in situ. On day 3, the number of osteoclasts on the appliance side of the experimental group was significantly increased compared with the control group (P < .05). On day 7, the number of osteoclasts on the non-appliance side of the experimental group was significantly increased compared with the control group (P < .05). This study indicates that oral administration of EPO can increase the number of osteoclasts and may accelerate orthodontic tooth movement. PMID- 15898374 TI - Functional unilateral posterior crossbite effects on mastication movements using axiography. AB - This prospective study investigated the influence of functional unilateral posterior crossbite on mastication movements. The study group included 16 patients (nine girls and seven boys) with functional unilateral posterior crossbite involving three or more posterior teeth. A control group comprised 15 individuals (nine girls and six boys) with normal occlusion and the mean age of both groups was 9.17 years. The mandibular masticatory movements were registered, using computer axiography, for 30 seconds during chewing to determine the preference side of chewing. The patterns of the first, third, and fifth chewing cycles were compared with the preference side to establish whether they would predict the chewing preference side. The extent of the maximal lateral and vertical displacements of the mandible during chewing were compared between study and control groups. This study found that overall the left side was the preferred mastication side in 43.7% of individuals in the study and 46.7% in the control group. There was no relationship between the side of the crossbite and the masticatory preference side (Mc Nemar test, P = .5). No correlation was present between the patterns of chewing movements in the first, third, or fifth cycles. Both study and control groups showed similar maximal lateral and vertical mandible displacement. PMID- 15898375 TI - Experimental force definition system for a new orthodontic retraction spring. AB - A new geometry of orthodontic retraction spring was experimentally studied through an electronic device (platform for measuring forces), using strain gauges that were adapted to cantilever beams. The sample consisted of 36 titanium molybdenum springs, divided into three groups of 12 springs each. The springs were produced with different cross sections of 0.016 x 0.022 inch and 0.017 x 0.025 inch and with different angles between the extremities (120 degrees and 130 degrees). The springs were adapted to the platform in three different positions so that the force system developed by them could be known (horizontal forces, vertical forces, alpha-beta moments, and moment-to-force ratio M:F). The analysis of factorial variance and the Tukey honestly significant difference test were applied to verify the differences between the averages caused by three possible variation sources and the interactions between them. Regression analysis was also performed to obtain the spring rate. The results show the interactions between the three geometric variables, force magnitudes, and also the spring rates, which are compatible with the ones mentioned in the literature related to the subject. The spring rate was within the levels that are appropriate for clinical use (varying from beta = 33.1 gf/mm to beta = 43.9 gf/mm). PMID- 15898376 TI - Anchorage effect of various shape palatal osseointegrated implants: a finite element study. AB - The purpose of this study was to compare the anchorage effects of different palatal osseointegrated implants using a finite element analysis. Three types of cylinder implants (simple implant, step implant, screw implant) were investigated. Three finite element models were constructed. Each consisted of two maxillary second premolars, their associated periodontal ligament (PDL) and alveolar bones, palatal bone, palatal implant, and a transpalatal arch. Another model without an implant was used for comparison. The horizontal force (mesial 5N, palatal 1N) was loaded at the buccal bracket of each second premolar, and the stress in the PDL, implant, and implant surrounding bone was calculated. The results showed that the palatal implant could significantly reduce von Mises stress in the PDL (maximum von Mises stress was reduced 24.3-27.7%). The von Mises stress magnitude in the PDL was almost same in the three models with implants. The stress in the implant surrounding bone was very low. These results suggested that the implant is a useful tool for increasing anchorage. Adding a step is useful to lower the stress in the implant and surrounding bone, but adding a screw to a cylinder implant had little advantage in increasing the anchorage effect. PMID- 15898377 TI - Effect of light-tip distance on the shear bond strengths of composite resin. AB - The purpose of this study was to assess the effect of light-tip distance on the shear bond strength and failure site of brackets cured with three different light curing units: a high-intensity halogen (Astralis 10, 10-second curing), a light emitting diode (LED, e-Light, six-second curing), and a plasma arc (PAC System, four-second curing). One hundred and thirty-five bovine permanent mandibular incisors were randomly allocated to nine groups of 15 specimens each. Stainless steel brackets were bonded with a composite resin to the teeth, and each curing light was tested at zero, three, and six mm from the bracket. After bonding, all samples were stored in distilled water at room temperature for 24 hours and subsequently tested for shear bond strength. When the three light curing units were compared at a light-tip distance of zero mm, the three lights showed no significantly different shear bond strengths. At light-tip distances of three and six mm, no significant differences were found between the halogen and plasma arc lights, but both lights showed significantly higher shear bond strengths than the LED light. When evaluating the effect of the light-tip distance on each light curing unit, the halogen light showed no significant differences between the three distances. However, the LED light produced significantly lower shear bond strengths at a greater light-tip distance, and the plasma arc lamp showed significantly higher shear bond strengths at a greater light-tip distance. In hard-to-reach areas, the use of PAC system is suggested, whereas the LED evaluated in this study is not recommended. PMID- 15898378 TI - Effects of chlorhexidine and povidone-iodine mouth rinses on the bond strength of an orthodontic composite. AB - The purpose of this study was to determine whether the application of two antibacterial mouth rinses to etched and unetched enamel affects the shear bond strength (SBS) of an orthodontic composite resin. Eighty-five lower human incisors were divided into five groups, ie, group 1: control group, no mouth rinse was used; groups 2 and 3: mouth rinses were applied to the intact enamel surface before etching; groups 4 and 5: mouth rinses were applied to the etched enamel. A bonding agent and a composite resin were applied to the teeth surface. For shear bond testing, the specimens were mounted in a universal testing machine, and an apparatus attached to a compression load cell was applied to each specimen until failure occurred. The data were analyzed using analysis of variance and Tukey honestly significance tests. Fracture modes were analyzed by Mann-Whitney U-test. There was no statistically significant difference between the SBS values of group 1 (31.64 +/- 3.62 MPa) and group 4-five experimental applications (P > or = .05). However, the SBS value of group 3 (36.56 +/- 5.95 MPa) was significantly larger than those of group 4 (30.00 +/- 4.97 MPa) and group 5 (30.26 +/- 7.30 MPa). In addition, no significant differences were observed between group 1 and groups 2 (34.33 +/- 7.26 MPa) and 3 (36.56 +/- 5.95 MPa) (P > or = .05). Because the application of chlorhexidine and povidone-iodine before acid etching did not cause any decrease in bond strength, it is advisable for use under the orthodontic resin composite to obtain an antibacterial effect or to prevent the risk of bacteremia. PMID- 15898379 TI - Effect of antimicrobial monomer-containing adhesive on shear bond strength of orthodontic brackets. AB - A new antibacterial and fluoride-releasing bonding system consists of a self etching primer that contains an antibacterial monomer and a bonding agent that contains sodium fluoride. This study was to determine the effect of using this new adhesive on the shear bond strength of orthodontic brackets. Forty molar teeth were randomly divided into two groups. Group 1 consisted of 20 teeth that were etched for 15 seconds with 35% phosphoric acid, washed with a water spray for 10 seconds, and dried to a chalky white appearance, and the sealant was applied to the etched surface. The precoated brackets were placed on the teeth and light cured. Group 2 consisted of 20 teeth that were etched with 35% phosphoric acid for 15 seconds as suggested by the manufacturer when bonding to intact enamel. The teeth were washed with a water spray for 10 seconds and dried to a chalky white appearance, and the primer containing antibacterial monomer was applied to the etched surface, left for 20 seconds, and sprayed with a mild airstream. The adhesive was applied to each tooth, and the precoated bracket was placed and light cured. There were no significant differences (P = .220) in the shear bond strengths of the two groups. The mean shear bond strength for the antibacterial fluoride-releasing adhesive was 11.7 +/- 5.6 MPa and for the control was 9.6 +/- 5.0 MPa. The use of an antibacterial fluoride-releasing adhesive system did not affect the shear bond strength of the orthodontic brackets within the first half hour after initial bonding. PMID- 15898380 TI - Influence of a nonrinse conditioner on the bond strength of brackets bonded with a resin adhesive system. AB - The objective of this study was to compare the effect of a nonrinse conditioner (NRC) and the conventional acid-etch technique on the shear bond strength and the adhesive remnant on the tooth after debonding brackets bonded with the resin orthodontic adhesive system Transbond XT. A total of 40 human premolars were divided into group I, phosphoric Acid/Transbond XT (n = 25) and group II, NRC/ Transbond XT (n = 15). Shear bond strength was measured with a universal test machine with a crosshead speed of 1 mm/min. The adhesive remnant was quantified using image analysis equipment. Scanning electron microscope (SEM) observations were also carried out to observe enamel surfaces treated with each product. No significant differences were observed in the bond strengths of the two groups evaluated. The amount of adhesive remnant on the tooth after debonding was significantly less when conditioning the enamel with NRC compared with phosphoric acid. SEM observations showed that the NRC produced a more conservative etch pattern than phosphoric acid. PMID- 15898381 TI - Sandblasted metal brackets bonded with resin-modified glass ionomer cement in vivo. AB - The aim of this in vivo study was to evaluate the effects of sandblasting metal brackets on their clinical performance when resin-modified, chemically cured glass ionomer cement was used for bonding. A total of 60 patients with a range of malocclusions were allocated randomly into two groups. For the first 30 cases, teeth were divided into quadrants so that sandblasted, mesh-based metal brackets (SB) were bonded directly to the upper left and lower right quadrants using the resin-modified glass ionomer cement. The mesh-based (no sandblasting) brackets bonded to the other quadrants with the same adhesive were used as control (CO). A split-mouth design was used, and the allocation of the brackets per quadrant was reversed for the second 30 cases. Sandblasting of the bracket bases was accomplished using 25-microm aluminum oxide particles for three seconds. The manufacturer's instructions were followed for bonding. The number, site, and date of first-time bracket failures were monitored throughout active orthodontic treatment, and the observation time was 20 months. The bond failure rates were 4.9% and 4.3% for the SB and CO brackets, respectively. No statistically significant difference was found between the groups for failure rates. The bond failure sites were predominantly at the enamel-adhesive interface in both groups. Sandblasting did not have a positive effect on the clinical performance of the mesh-based metal brackets when bonded with resin-modified glass ionomer cement. PMID- 15898382 TI - Shear bond strength, bond failure, and scanning electron microscopy analysis of a new flowable composite for orthodontic use. AB - A new dental flowable composite, Denfil Flow, was evaluated for the bonding of orthodontic brackets by determining its shear bond strength (SBS) and the mode of bond failure after debonding. Eighty extracted human premolars were divided into two equal groups. Metal brackets were bonded to etched enamel using a composite resin control (Transbond XT) or Denfil Flow. After 72 hours of incubation in saline solution at 37 degrees C, debonding was performed with a shearing force. The SBS and the mode of bond failure were examined. In addition, representative samples from each group were examined by scanning electron microscopy (SEM). No significant difference was observed in the SBS between the groups, and a clinically acceptable SBS was found for the two adhesives. Bond failures occurred mostly in the bracket-adhesive interface, without significant differences between the groups. At SEM analysis, Denfil Flow showed a greater frequency of air bubbles within the resin than did Transbond XT. In conclusion, Denfil Flow displayed the same SBS as traditional composite resins and similar bond failures. Further clinical studies are required. PMID- 15898383 TI - Long-term clinical outcome of rapid maxillary expansion as the only treatment performed in Class I malocclusion. AB - The purpose of this study was to investigate the long-term clinical responses of rapid maxillary expansion as the only treatment performed in Class I malocclusion using the Haas-type appliance. The longitudinal sample consisted of 90 sets of study models from 30 consecutive patients (12 males and 18 females) selected on the basis of the following inclusion criteria: all patients (1) had a Class I malocclusion with transverse maxillary/mandibular skeletal discrepancies, (2) were treated nonextraction in the early/ mid mixed dentition, (3) presented with mandibular dental arches with mild or no crowding, and (4) had no subsequent comprehensive orthodontic treatment implemented in either the maxilla or the mandible. The mean age was 8.2 years when treatment was initiated. Treatment outcomes were evaluated at pretreatment A1, short-term follow-up (one year after A1) A2, and long-term follow-up (four years after A2) A3. The changes in maxillary arch width and arch length were quantified and compared among assessment stages A1, A2, and A3 using the Student's t-test. The results demonstrated a highly significant increase in maxillary arch width in both the short- and long-term follow-ups. The arch width increased significantly during treatment and decreased slightly during the long-term follow-up. The long-term clinical response demonstrated the efficacy and stability of this type of treatment in achieving maxillary arch width. The follow-up examination during the early/mid/permanent dentition confirmed the validity of overtreatment. PMID- 15898384 TI - Friction of orthodontic elastomeric ligatures with different dimensions. AB - The aim of this study was to evaluate in vitro the effect of variations in the size of elastomeric ligatures on the static frictional resistance generated by orthodontic sliding mechanics under dry condition. Frictional forces generated by elastomeric ligatures treated with a lubricating material (silicone) were analyzed as well. An Instron testing machine was used to assess the static frictional forces of a 0.019 x 0.025-inch stainless steel rectangular wire that was ligated to a molar convertible tube and to three stainless steel 0.022-inch pre-adjusted brackets with elastomeric ligatures with different dimensions: small, medium, and large. The static friction produced by two prototypes of silicone-lubricated elastomeric ligatures was also measured. The small and medium elastomeric ligatures produced significantly less friction than the large ligatures. No statistically significant difference was found between small and medium ligatures. The decrease in frictional forces of small and medium modules had to be ascribed mainly to the smaller thickness of both ligatures with respect to large ligatures. The lubricated elastomeric ligatures generated significantly smaller frictional forces than nonlubricated elastomeric ligatures with different dimensions. The variation in the dimensions of the elastomeric ligatures is able to influence the static frictional resistance generated by orthodontic sliding mechanics in the buccal segments. The use of small and medium elastomeric ligatures determines a 13-17% decrease in static friction compared with large ligatures. Silicone-lubricated modules can reduce static friction by 23-34% with respect to the small and medium nonlubricated elastomeric ligatures and by 36-43% compared with nonlubricated large ligatures. PMID- 15898385 TI - Skeletal, dental and soft-tissue changes induced by the Jasper Jumper appliance in late adolescence. AB - The purpose of this study was to evaluate the skeletal, dental, and soft-tissue changes in late-adolescent patients treated with Jasper Jumpers applied with sectional arches. The study sample consisted of 30 subjects (15 treated, 15 untreated) with skeletal and dental Class II malocclusion. Our study was carried out on 75 lateral cephalometric films. Among these radiograms, 15 were taken before the leveling stage in the treatment group. Half of the remaining 60 were taken before placement and after removal of the Jasper Jumper appliance in the treatment group and the other half at the beginning and six months after in the control group. The patient selection criteria were Class II malocclusion caused by retrognathic mandible, normal or low-angle growth pattern, and postpeak growth period. The statistical assessment of the data suggests that the sagittal growth potential of the maxilla was inhibited. There were no significant changes in the vertical skeletal parameters. The mandibular incisors were protruded and intruded, whereas the maxillary incisors were retruded and extruded. The upper molars tipped distally as the lower molars tipped mesially. Because of these changes, the occlusal plane rotated in the clockwise direction. Overbite and overjet were reduced, and the soft-tissue profile improved significantly. The results revealed that, in late-adolescent patients, the Jasper Jumper corrected Class II discrepancies mostly through dentoalveolar changes. It is suggested that this treatment method could be an alternative to orthognathic surgery in borderline Class II cases. PMID- 15898386 TI - Anchorage provided during intra-arch distal molar movement: a comparison between the Nance appliance and a fixed frontal bite plane. AB - The aim of this retrospective study was to evaluate and compare the anchorage provided with the Nance appliance (NA) and the fixed frontal bite plane (FBP) during intra-arch distal molar movement. After a sample size calculation, 20 patients were recruited and randomly selected for each group from patients who fulfilled the following criteria: use of an intra-arch Ni-Ti coil appliance with either NA or FBP to provide anchorage during a six-month molar distalization period, no orthodontic treatment before molar distalization, and first and second maxillary molars in occlusion. The outcome measures assessed were anchorage loss, ie, anterior movement of maxillary central incisors, distal movement of maxillary molars, and bite opening effect. The mean age in the NA group was 14.7 years (SD 1.09) and in the FBP group 15.0 years (SD 0.99). The data revealed that the maxillary central incisors moved anteriorly 1.4 mm in the NA group and 1.9 mm in the FBP group. The difference in anchorage loss was not significant. The mean amount of molar distalization within the maxilla was 1.7 mm in the NA group and 1.8 mm in the FBP group. In both groups, the overbite was significantly reduced and the overbite was decreased significantly more in the FBP group. Because neither the NA nor FBP provided stable anchorage, a second treatment phase is recommended to reverse the anchorage loss after distal molar movement. If molar distalization is planned in deep bite cases, the FBP is the anchorage system of choice. PMID- 15898387 TI - A mini-implant for orthodontic anchorage in a deep overbite case. AB - This article describes the orthodontic treatment of a 19-year-old female patient with anterior crowding. There was a moderate arch length discrepancy in the lower dental arch, a significant deep overbite, and a "gummy smile." We inserted an orthodontic mini-implant as anchorage for the intrusion of the upper incisor segment, followed by alignment of the upper and lower dental arches with an edgewise appliance without tooth extraction. The overbite was corrected from +7.2 mm to +1.7 mm by upper incisor intrusion, and the gummy smile was improved. Good occlusion and facial esthetics were achieved, and these results have been maintained for two years after completion of the active treatment. PMID- 15898388 TI - Use of onplants as stable anchorage for facemask treatment: a case report. AB - A hexagonal onplant of 7.7 mm diameter was placed on the palatal bone of the maxilla in an 1-year five-month-old female patient with a Class III malocclusion and midface deficiency. Elastic traction (400 g per side) was applied from a facemask to the onplant at 30 degrees to the occlusal plane 12 hours per day for 12 months. The maxilla was found to have displaced forward and downward by 2.9 mm. The mandible was rotated downward and backward. There was a 3 degrees increase in mandibular plane angle and an increase in the lower face height. Clinically, there was a significant improvement in midface esthetics, noted by an increase in fullness of the infraorbital region and correction of the skeletal discrepancy between the maxillary and mandibular jaw relationship. Contrary to the reports that use teeth rather than onplants as anchorage, there was no forward movement of the maxillary molars and minimal extrusion of the maxillary molars. These results suggest that onplants can be used as an extremely stable anchorage for maxillary orthopedic facemask treatment. PMID- 15898389 TI - Abnormal mandibular growth after craniovertebral surgery in Morquio syndrome type A. AB - Morquio syndrome or MPS4A is an autosomal recessive inherited metabolic disease, due to a deficiency of N-acetil-galactosamine-6-sulfatase (OMIM 253000). Hypoplastic odontoid processes causing atlantoaxial subluxation and cervical myelopathy are usual clinical findings. Surgical intervention of craniocervical fusion is often performed to prevent this complication. Clinical and cephalometric findings in a patient affected by Morquio syndrome after craniovertebral surgery are described. Facial growth pattern in the lateral plane changed dramatically. The mandibular gonial angle (ArGoMe), the body of the mandible (GoGn), and the total length of the mandible (CoGn) increased abnormally, whereas the mandibular ramus (CoGo) exhibited normal growth. Knowledge of the possibility of abnormal mandibular growth may contribute in long term orthodontic management of such subjects. PMID- 15898390 TI - An adult case of skeletal open bite with a large lower anterior facial height. AB - Control of the height of posterior dentoalveolar regions is of great importance for the correction of skeletal open bite. Traditionally, second premolar extraction facilitates the closure of open bite by inducing a counterclockwise mandibular rotation without molar intrusion. This article reports treatment for a 24-year six-month-old female patient with an open bite and large anterior facial height. She complained of occlusal disturbances and difficulty of lip closure because of the open bite. Overjet and overbite were +3.0 mm and -3.0 mm, respectively. To correct open bite and crowding, the bilateral extraction of the maxillary and mandibular second premolars plus multibracket appliances for mesial movement of the molars was selected as the treatment plan. After a two-year treatment, an acceptable occlusion was achieved, the lower anterior facial height was decreased, and the lips showed less tension in a lip closure. An acceptable occlusion was maintained without recurrence of the open bite during a three-year retention period, indicating a long-term stability of the occlusion. The results of this treatment indicated that the correction of open bite with no or less molar intrusion or incisor extrusion is of great importance for achieving stable occlusion and avoiding the relapse of open bite. PMID- 15898391 TI - Nonextraction treatment of upper canine-premolar transposition in an adult patient. AB - This article reports the successful treatment of a unilateral maxillary canine and first premolar transposition without the extraction of the premolar in an adult patient. A female patient, 21 years and three months of age, had moderate crowding in the upper arch with complete transposition of the canine and first premolar. After distal movement of the upper molars with a lingual arch and headgear appliance, the upper left first premolar and canine were transposed. Thirty-eight months after the placement of preadjusted appliances, the transposed canine and premolar were reordered in the proper positions. The total active treatment period was 49 months. After two years of retention, the occlusion is generally stable. PMID- 15898392 TI - Winged maxillary central incisors with unusual morphology: a unique presentation and early treatment. AB - Winged incisors are a well-recognized clinical finding. In this report, the disorder is briefly reviewed and a unique case of winging of the two maxillary central incisors having unusual morphology in an eight-year six-month-old boy is presented. The two winged maxillary central incisors were derotated using an anterior sectional wire inserted into a pair of twin brackets, one bonded to each of the two central incisors, and reciprocal anchorage. PMID- 15898393 TI - Zygomatic anchorage for en masse retraction in the treatment of severe Class II division 1. AB - An adult female patient who presented with a severe Class II division 1 malocclusion was treated by en masse retraction of upper anterior teeth against zygomatic anchorage. This case report describes the surgical and orthodontic procedures followed during the treatment. En masse retraction of the six anterior teeth by using zygomatic bone anchorage proved to be an efficient method for the correction of a severe overjet problem. PMID- 15898394 TI - Evidence-based journalism. PMID- 15898395 TI - Re: Amount and direction of temporomandibular joint growth changes in Herbst treatment: a cephalometric long-term investigation. Angle Orthod. 2003;73(5):493 501. PMID- 15898396 TI - GPOs begin issuing GLNs as standard location IDs. PMID- 15898397 TI - Declines seen in gown prices. PMID- 15898398 TI - In Secours finds ways to cut drug costs. PMID- 15898399 TI - The relationship between managerial leadership behaviors and staff nurse retention. AB - The purposes of this study were to describe perceptions of managerial leadership behaviors associated with staff nurse turnover and to compare nurse manager leadership behaviors as perceived by managers and their staff nurses. Effective leadership styles among nurse managers have been associated with staff nurse job satisfaction and retention. Although both transformational and transactional leadership styles have been described as effective, it is unclear which nurse manager leadership behaviors contribute most to staff nurse retention. This descriptive, correlational study was conducted at a 465-bed community hospital in the northeastern United States. All staff nurses and nurse managers employed in both ambulatory and acute care nursing units were invited to participate in the study. The study sample comprised 79 staff nurses and 10 nurse managers, who completed demographic forms and the 45-item Multifactor Leadership Questionnaire, which measures 12 dimensions of leadership style. Data were collected from July through September 2003. Active management by exception as perceived by staff nurses was the only managerial leadership style associated with staff nurse turnover (r = .26, p = .03). Compared with the perceptions among their staff nurses, nurse managers consistently perceived that they demonstrated a higher mean frequency of transformational leadership behaviors. The transactional leadership style of active management by exception not only appeared to be a deterrent to staff nurse retention but also reflected leadership perceptions among staff nurses who work evening and night shifts. This study also provides further evidence regarding a trend in which nurse managers and staff nurses do not concur on the frequency of transformational leadership behaviors but do demonstrate agreement on the frequency of transactional leadership behaviors. PMID- 15898400 TI - Patients' perceptions of pain management and use of coping strategies. AB - As a result of the Joint Commission on Accreditation of Healthcare Organizations accreditation standards for 2001, pain management for hospitalized patients has become a top priority of healthcare facilities. In addition to using the traditional pharmacological approach to pain management, many patients also use complementary or alternative medicine (CAM) treatments. However, CAM treatments may not be discussed or offered to patients by healthcare providers who lack awareness about these alternatives. The purpose of this study was to assess patients' perceptions of pain, their beliefs about the use of pharmacological and CAM pain management techniques, and their satisfaction with pain management. Researchers verbally administered a survey to patients by using a combination of open-ended questions and a 0-10 rating scale, and they recorded their responses. This study was conducted in a not-for-profit teaching hospital in the southeast United States. Convenience sampling was used to select the 137 patients who completed the surveys. No treatment intervention was provided. Pharmacological treatment was the primary method expected and used by the majority of patients for pain management. Chi-squared statistics were used to analyze nonparametric data. An analysis of variance was used to analyze parametric data. The frequency with which nonpharmacological CAM options were used ranged from 6 to 34 percent. The most commonly used CAM method was distraction, such as watching television or reading. PMID- 15898401 TI - Adapting to change in healthcare: aligning strategic intent and operational capacity. AB - The concept of a stakeholder is commonplace in a business context. Participative- democratic communication refers to those organization-wide principles and practices that "represent" many relevant stakeholders in the decision making of work-related activities. One case in point is as follows: In May 2000, the Spartanburg Regional Healthcare System board of directors forced its CEO to resign, ending his 6-year tenure and the heated controversy of the previous 2 months. The former CEO focused primarily on strategic growth to the detriment of operations. Through participative-democratic practices, the interim CEO mended the damaged relations between the hospital administration, the community, and employee stakeholders in surfacing conflict to bolster operational efficiency. The current CEO attended to building stakeholder relationships and trust as a way to wed strategic growth and the organization's capacity to maintain it. Top executive managers and directly involved community political leaders helped in developing the participative-democratic communication principles set forth in this article. These core principles are (a) creating the space for new communicative interaction, (b) safeguarding a credible and open process, and (c) reclaiming suppressed views. PMID- 15898402 TI - The "New" medical malpractice crisis--part 2. PMID- 15898403 TI - Health professionals beware of prescription pain medication abuse and diversion. PMID- 15898404 TI - Letter from the editor: lung cancer and gender bias. PMID- 15898405 TI - Case of the season: Kaposi sarcoma. PMID- 15898406 TI - Epidemiology of lung cancer. PMID- 15898407 TI - The 2004 World Health Organization classification of lung tumors. PMID- 15898408 TI - The imaging manifestations of lung cancer. AB - Imaging plays an integral role in diagnosing, staging, and following patients with lung cancer. Many lung neoplasms are detected on chest radiographs, but the majority of patients have advanced stage disease at the time of presentation. There is a wide spectrum of radiologic manifestations of lung cancer, and recognition of these findings is essential for patient management. As we continue to understand more about tumor biology, new imaging techniques likely will emerge. Nevertheless, the chest radiograph and CT remain important tools in establishing the diagnosis of lung cancer. PMID- 15898409 TI - Multidetector CT of the solitary pulmonary nodule. PMID- 15898410 TI - CT, MR, and PET imaging in staging of non-small-cell lung cancer. PMID- 15898411 TI - State-of-the-Art FDG-PET imaging of lung cancer. AB - The D-glucose analog 2-(fluorine-18)-fluoro-2-deoxy-D-glucose (FDG) is the most commonly used radionuclide in positron emission tomography (PET) of lung cancer. FDG-PET is a molecular imaging technique that images the preferential accumulation of FDG in malignant tissues with increased metabolism. Although FDG PET is sensitive in the detection of lung cancer, FDG is not tumor specific and may accumulate in a variety of nonmalignant conditions. Occasional false-negative results may also occur. Whole body FDG-PET is a useful noninvasive technique to stage known or suspected non-small-cell lung cancer. The results allow more efficient use of invasive methods for histopathological staging. The combined use of CT and PET in dual imaging increases the number of patients with correctly staged non-small-cell lung cancer. CT/PET is also useful in the assessment of recurrent or residual disease. Future imaging agents are being developed which may allow more selective accumulation of radiopharmaceutical in malignant tissues. PMID- 15898412 TI - Percutaneous biopsy in lung cancer. PMID- 15898413 TI - Image-guided radiofrequency ablation as a new treatment option for patients with lung cancer. PMID- 15898414 TI - Imaging for lung cancer restaging. AB - Diagnostic imaging plays an important role in the monitoring of tumor response during lung cancer restaging to evaluate the efficacy of chemotherapy and/or radiation therapy during treatment, and in the detection of recurrent or metastatic neoplasm after treatment has been completed. While CT represents the primary imaging modality for lesion evaluation during restaging and for surveillance imaging once therapy has been completed, studies evaluating the role of 18-fluoro-2 deoxyglucose positron emission tomography (FDG-PET) in lung cancer restaging have shown promise regarding the detection of residual and recurrent neoplasm, and in evaluating for early response to first line therapy. With both CT and FDG-PET, residual or recurrent disease should, when possible, be differentiated from therapy-related changes in the lungs. We review the role of imaging in lung cancer restaging with attention to CT and FDG-PET for treatment assessment and the detection of recurrent or metastatic disease. PMID- 15898415 TI - CT screening for lung cancer. PMID- 15898416 TI - The world of Chef Jorge. PMID- 15898417 TI - [Effect of nucleolin down-regulation on the proliferation and apoptosis in C2C12 cells]. AB - OBJECTIVE: To clarify the effect of nucleolin on the proliferation and apoptosis in C2C12 cells. METHODS: After inhibiting the expression of nucleolin using antisense oligonucleotides, the cellular proliferation was determined by MTT, and the apoptosis was detected by flow cytometry (FCM) assays and DNA ladder assays. RESULTS: After being transfected with antisense oligonucleotides for 24 hours, Western blotting showed that the expression of nucleolin was repressed significantly. In cells treated with antisense oligonucleotides, the cellular proliferation was obviously inhibited; the apoptotic cell increased significantly; and the "DNA ladder" was clearly observed. But the sense and random oligonucleotides had no effect on the cellular proliferation and apoptosis. CONCLUSION: The down-regulation of nucleolin can inhibit the cellular proliferation and initiate the apoptosis in C2C12cells. PMID- 15898418 TI - [Comparison of 3 working definitions of metabolic syndrome in male medical examinees]. AB - OBJECTIVE: To compare the prevalence of the metabolic syndrome (MS) using 3 working definitions proposed respectively by the World Health Organization (WHO, 1999) , the Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults ( ATP III, 2001 ), and the Chinese Diabetes Society ( CDS, 2003). METHODS: MS was diagnosed in 739 male medical examinees by the 3 working definitions respectively, then the prevalence and the concordance of 3 working definitions was compared. RESULTS: Among 739 participants the prevalence was 36.9% by the WHO definition, 11.8% by the ATP III definition and 21.0% by the CDS definition. Among all the testees 68.6% were classified as either having or not having the MS under the 3 definitions. The consistency in the diagnosis of MS was 72.5% by the WHO definition and the ATP III definition, 81.2% by the WHO definition and the CDS definition, and 83.5% by the ATP III definition and the CDS definition. The prevalence of insulin resistance was the highest among the components of the WHO definition. The prevalence of hypertension was the highest while the prevalence of obesity was the lowest by the ATP III definition. Among the components of the CDS definition, the prevalence of obesity was the highest. The fasting insulin and insulin resistant index (HOMA-IR) were both significantly higher in the MS subjects than that in the non-MS subjects. CONCLUSION: A universally accepted definition of the metabolic syndrome is needed. PMID- 15898419 TI - [Determination of HLA-DRB1 gene polymorphism in Luoba ethnic group of Tibet]. AB - OBJECTIVE: To investigate the frequencies of allele and genotype of HLA-DRB1 gene in Luoba ethnic group. METHODS: HLA-DRB1 genes from 92 individuals of Luoba ethnic group in Tibet Autonomous region were investigated, using PCR-SSO (polymerase chain reaction -sequence specific oligo-nucleotide) genotyping methods. We compared allele frequencies of Luoba population in HLA-DRB1 locus with that of various ethnic groups distributed worldwide, constructed the phylogenetic tree by UPGMA (unweighted pair group method with arithmetic mean), and counted genetic distance by Nei measure. RESULTS: Of the 11 DRB1 alleles detected, the two most common genes were DRBI * 04( gene frequency: 27.20% ) and DRB * 12 (25.50%) , which covered 52.70% of the total alleles detected from Luoba ethnic group. The frequencies of HLA-DRB1 * 14(15.20% ), DRB1 * 15(9.80% ) and DRB1 * 08(8.20% ) were more than 5%. Luoba was unique in the distribution of HLA alleles. The genetic distance between Luoba and Tibetan was closer than that with other population. A dendrogram based on the DRB1 genes by cluster analysis suggested that Luoba ethnic group might cluster with Tibetan firstly, then cluster with other population living in China (except Uygr) , and cluster with Caucasian and Black finally. CONCLUSION: The HLA-DRBI gene frequency of Luoba individuals in Tibet Linzhi had some differences compared with that of other Chinese population. The kindred relation between Luoba and Tibetan was closer than that with other population, which was coincident with the results of ethnology, history and sociology. PMID- 15898420 TI - [Expression of exogenous gene in bone marrow stromal cells by liposome]. AB - OBJECTIVE: To transform eukaryotic expression vector pEGFP-PDX-1 into marrow stromal cells by liposome and to optimize the conditions of transformation. METHODS: The recombinant vector was identified by enzyme digestion analysis and sequencing. The recombinant plasmid was transformed into bone marrow stromal cells and it changed the quantity of DNA or liposome. The expression of PDX-1 gene in the transformed cells was detected by immunocytochemical staining. RESULTS: Enzyme digestion analysis and sequencing showed that the interesting gene was integreted into the recombinant vector. We obtained satisfactory efficiency of transfection when the ratio of DNA and liposome was 1 : 1 or 1 : 2. The PDX-1 in the transformed cells was expressed by immunocytochemical staining. CONCLUSION: The eukaryotic expression vector pEGFP-PDX-1 was constructed for the first time in China. We have enhanced the efficiency of transfection by optimizing the transformation conditions. It is possible to use the bone marrow stromal cells as seed cells in tissue-engineering. PMID- 15898421 TI - [Effect of baicalin on the proliferation of insulinoma cell line]. AB - OBJECTIVE: To investigate the effect of baicalin on the proliferation of insulinoma cell line and the molecular mechanism involved. METHODS: Such methods as light microscope, MTT assay, flow cytometry and Western blotting were applied to investigate the effects of baicalin (0, 100, 200, and 400 microg/ml baicalin treated for 24 h or 200 microg/ml baicalin treated at different time points) on the cell proliferation, cell survival rate, the cell cycle and related molecular mechanisms. RESULTS: The number of proliferating cells obviously decreased with the increase of baicalin under the light microscope, and the survival rate of cells decreased as determined by MTT assay. After being treated with baicalin, the number of insulinoma cells in S-phase obviously decreased from 38.2% (0 microg/ml) to 9.4% (400 microg/ml), and the number of cells in phase G1 increased from 56.4% (0 microg/ml) to 85.9% (400 microg/ml). In the meantime, the expression of cyclin D1 was obviously declined by Western blotting. CONCLUSION: Baica-lin can inhibit the proliferation of insulinoma cells, and the down regulation of the expression of cyclin D1 might also be involved in these events. PMID- 15898422 TI - [Effect of ginsenoside on the cellular proliferation, apoptosis and cell cycles in LC A549 and HUVEC 304 cell lines]. AB - OBJECTIVE: To determine the effect of ginsenoside on the cellular proliferation, apoptosis and cell cycles in LC A549 and HUVEC 304 cell lines. METHODS: A549 and HUVEC 304 cell lines were cultured with different concentrations of ginsenoside. Cellular proliferation was detected with MTT, apoptosis and cell cycles were checked with Flow Cytometer, and change of microstructure was observed by transmission electron microscope. RESULTS: The apoptosis rate was 29.8% in A549 cell lines after being interfered with ginsenoside at 3 x 10(-6) mol/L, significantly higher than that in the control group ( P < 0.05). No change was observed in the cell cycles after being interfered with ginsenoside. The inhibitive rate of ginsenoside was 12.53% for HUVEC 304 cell line at 1 x 10(-4) mol/L (P < 0.05 ). The cells induced by conditioned medium could be inhibited by ginsenoside, and apoptotic body could be found in cells induced by conditioned medium at 10(-6) mol/L. CONCLUSION: The proliferation of vascular endothelial cell could be inhibited by ginsenoside, and apoptosis could also be found in both tumor cells and cells induced by conditioned medium after being interfered with ginsenoside. PMID- 15898423 TI - [Effects of naoyian serum on VEGF protein expression in cultured rat cerebral microvascular endothelial cell with hypoxia]. AB - OBJECTIVE: To determine the effects of naoyian (NYA) serum on the expression of vascular endothelial growth factor (VEGF) protein in cultured rat cerebral microvascular endothelial cell (RCMEC) with hypoxia. METHODS: NYA serum was separated from rat heart which had been filled stomach with NYA successively for 3 days. The rat cerebral microvascular endothelial cells were taken from the Sprageu-Dawley rat brain at postborn 7 days. The rat cerebral microvascular endothelial cells were incubated at anaerobic incubator to establish the hypoxia models. The vigo of RCMEC was determined by MTT. The level of expression of VEGF protein was measured by cell immunohistochemistry and Western blot. RESULTS: The OD value of NYA serum group was higher than the control groups after hypoxia for 18 hours. VEGF protein was increased by hypoxia in cerebral microvascular endothelial cells (P < 0.05). The content of VEGF protein in NYA serum containing medium was more significantly elevated than those cultured in other control media (P < 0.01). CONCLUSION: VEGF protein was induced by hypoxia in rat cerebral microvascular endothelial cells, and NYA could upregulate the expression of VEGF protein, which may be one of the protection mechanisms for cerebral microvascular endothelial cells. PMID- 15898424 TI - [Effect of intrathecal pumping morphine on immunological function in rats with formalin pain]. AB - OBJECTIVE: To evaluate the immunological function in rats with formalin inflammatory pain through intrathecal pumping different dosages of morphine. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into 4 groups (n = 8 in each group): saline group (NS) and morphine group included M1 group (10 microg/h) , M2 group (5 microg/h), and M3 group (2.5 microg/h). Chronic intrathecal catheterization was performed under anesthesia with 10% chloral hydrate (300-350) mg/kg according to M2 group (5 microg/h) and M3 group (2. 5 microg/h). Chronic intrathecal catheterization was modified Yaksh's. After 7 days, pain intensity scoring (PIS) was utilized to assess antinociceptive effect of morphine. And spleens were aseptically removed to obtain splenic cells. T lymphocyte function was evaluated based on Concanavalin-A induced splenocyte proliferation. A modified lactic acid dehydrogenase release assay was used to assess NK cell activity. Phenotypic expression of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+ / CD8+ ) and NK cell ( CD161+) in the spleen were analyzed by flow cytometry. RESULTS: Compared with the NS group, PIS of morphine group decreased obviously (P < 0.01) and was dose dependent in the early and late phase of formalin pain, but there were no significant differences among morphine groups. Spleen index, splenocyte proliferation and NK cell activity were significantly suppressed by intrathecal pumping morphine. Phenotypic expression of T lymphocyte subsets and NK cell assessed by flow cytometry were different from the control group in all morphine groups. CONCLUSION: There was significant antinociception of intrathecal pumping morphine. After intrathecal pumping different dosages of morphine (10 microg/h,5 microg/h, and 2.5 microg/h), the function of cellular immunity was suppressed and was dose-dependent. PMID- 15898425 TI - [Comparision of several different peritoneal fibrosis rat models]. AB - OBJECTIVE: To compare different types of peritoneal fibrosis models in rats. METHODS: Thirty-four SD rats were divided into 5 groups: control group (Group 1), normal saline group (Group 2), high glucose group (4.25% peritoneal dialysate, 4.25% PDF, Group 3), high glucose + lipopolysaceharides (LPS) group (4.25% PDF + LPS, Group 4), high glucose + erythromycin group (4.25% PDF + lactobionate erythromycin, Group 5). A 2-hour peritoneal equilibration test (PET) was performed after 5 weeks. Then animals were humanely killed. Dialysate-to-plasma urea ratio (D/ Purea), glucose reabsorption (D2/D0), net ultrafiltration (UF) volume were determined. The level of fibronectin in peritoneal tissues was measured by immunohistochemical method. Peritoneal membrane histology was evaluated by light microscopy. RESULTS: The D2/D0 ratio and net ultrafiltration volume in Groups 3, 4, and 5 were significantly lower than those in Groups 1 and 2 (P < 0.05) . The D/Purea ratio in Groups 3, 4 and 5 were significantly higher than that in Groups 1 and 2 (P < 0. 05 ). The level of fibronectin in Groups 3, 4 and 5 were significantly higher than that in Groups 1 and 2 (P < 0.05 ). CONCLUSION: Different types of peritoneal fibrosis models in rats has been established. The best model is high clusion glucose + erythromycin. PMID- 15898426 TI - [Cloning and characterization of new genes of Schistosoma japonicum]. AB - OBJECTIVE: To clone and characterize new genes of Schistosoma japonicum, Sj, and to provide efficient vaccine candidates. METHODS: Sj adult cDNA library was screened with rabbit sera raised against male worm soluble antigen. The inserted cDNA fragments from the positively selected clones were amplified with PCR and further sequenced, as well as characterized through internet NCBI GenBank software. RESULTS: Eleven positive clones were obtained and two were verified by GenBank as new, including a novel gene designated as Sj-P8 (GenBank accession No. AF517843) and a new partial cDNA of Sj myosin (GenBank accession No. AY770506). The two new genes encoded a transmembrane protein of 75 amino acids and a myosin protein fragment of 212 amino acids respectively. CONCLUSION: The newly obtained genes may provide useful information for the research on Sj vaccine. PMID- 15898427 TI - [CT appearance and p16 gene abnormality of peripheral lung cancer]. AB - OBJECTIVE: To investigate the relationship between the mutation and abnormal expression of p16 gene in peripheral lung carcinoma and its CT manifestations. METHODS: Immunohistochemistry and PCR-SSCP were used to detect P16 protein expression and p16 gene mutation of 52 cases of peripheral lung cancer. All patients were scanned with spiral CT before the operation and proved by pathology. RESULTS: Of the 52 cases of lung cancer tissues, the negative expression rate of p16 gene protein was 53.8% (28/52), and the deletion or mutation rate of the exon 2 was 23.1% (12/52). There were significant statistical differences of p16 gene defect and its protein loss rates among groups of different clinical stages (P < 0.05), but among groups of different tissue types, different differentiation degree p16 gene defect and its protein loss rates showed no significant statistical difference (P > 0.05). In lung cancer patients with CT appearances of thin spicule, speculated protuberance, pleural indentation, and metastasis of lymph node, p16 gene and its protein loss rates were much higher than those without CT manifestations mentioned above (P < 0.05). However, there were no statistical differences among groups of different tumor sizes, with or without lobulation, with or without cavity, and different contrast enhanced CT values (P > 0.05). CONCLUSION: p16 gene mutation and abnormal expression may play an important role in the occurrence and development of lung cancer, and it is relative to CT appearances of lung cancer. p16 gene may be used as a predicting index for clinical diagnosis and prognosis assessment. PMID- 15898428 TI - [Immunogenicity of bovine jugular vein conduits treated with different cross linking methods]. AB - OBJECTIVE: To evaluate the immunogenicity of bovine jugular vein conduits (BJVCs) treated with different cross-linking methods. METHODS: The BJVCs were treated with glutaradehyde (GA), dye-mediated photooxidation (DMP) and polyepoxy compound (PC) (n = 10). The tissue homogenates obtained from BJVCs treated with PC, GA, DMP, and fresh BJVCs, were mixed with the complete Freand adjavant to form the emulsive antigen, which were used to immunize rabbits correspondently. The antibody concentrations to BJVCs in those rabbits' serum were measured by counter double immuno diffusion. The immunologic responses to the BJVCs in different groups were measured with Western blotting. RESULTS: The positive bands appeared when the sera of rabbits were immunized by fresh BJVCs reacted with antigens of fresh BJVCs, but no bands appeared when the sera of rabbits were immunized by fresh BJVCs reacted with those antigens of the BJVCs treated with GA, DMP, and PC in Western blotting. CONCLUSION: The immunogenicity of BJVCs treated with PC, DMP, and GA can be reduced significantly and meet the clinical standard. PMID- 15898429 TI - [Pathway for macrophage invasion into the tunica media and the intima in vein graft]. AB - OBJECTIVE: To test whether the macrophages in vein grafts may invade into the vascular wall from the adventitia apart from the lumen. METHODS: Expression of CD68 (a marker of macrophage) and CD31 (a marker of endothelial cells) in 30 vein grafts were detected by confocal immunofluroscence. Images were processed with Silicon Graphics Octane. RESULTS: In normal veins, there were a few CD68 positive cells in the adventitia, but few CD68 positive cells were found in the media and the intima. In the diseased vein, CD68 positive cells were significantly increased and present in all layers of the vascular wall, the adventitia, media and intima, extending from the adventitia to the media. There was a close relationship between the penetration of CD68 positive cells into the media from the adventitia and the angiogenesis in the media. CONCLUSION: The adventitia may be an important pathway for macrophage invasion. PMID- 15898430 TI - [Expression of bcl-2 and bax protein in uterine leiomyosarcomas and leiomyomas]. AB - OBJECTIVE: To determine the expression and significance of the apoptosis regulatory protein bcl-2 and bax in uterine leiomyosarcomas and uterine leiomyomas. METHODS: The expression of bcl-2 and bax were examined by immunohistochemical ABC staining with specific antibodies in 34 cases of uterine leiomyosarcomas (18 cases in stage I , 1 in stage II , 8 in stage III, and 7 in stage IV ), 34 uterine leiomyomas and 34 normal myometrium samples. RESULTS: Bcl 2 expression was higher in uterine leiomyomas than that in normal myometrium and in uterine leiomyosarcoma (P < 0.01 ); bcl-2 expression in uterine leiomyosarcoma was higher than that in normal myometrium (P < 0.05). Bax expression in normal myometrium was higher than that in leiomyomas and in uterine leiomyosarcoma (P < 0.01). There was no obvious difference in bax expression between leiomyomas and leiomyosarcoma (P > 0.05). Both the expressions of bcl-2 and bax proteins in stage I - II were higher than those of in stage III - IV (P < 0.01). The bcl-2 positive leiomyosarcoma has a favorable clinical outcome than that of bcl-2 negative. CONCLUSION: The imbalance between the expression of bcl-2 and bax may play a certain role in the genesis of the uterine leiomyomas and leiomyosarcomas. Bcl-2 and bax may be involved in the early stage of the tumor genesis of uterine leiomyosarcoma. Bcl-2 might correlate with the prognosis of the uterine leiomyosarcoma. PMID- 15898431 TI - [Maternal serum matrix metalloproteinase-9 and interleukin-6 in predicting chorioamnhionitis in patients with premature rupture of membranes]. PMID- 15898432 TI - [Expression of cyclooxygenase-2 in fetal membranes of premature delivery]. AB - OBJECTIVE: To investigate the effect of cyclooxygenase-2 (COX-2) in pathogenesis of premature delivery. METHODS: A cross-sectional study was conducted with 30 women in the following categories: preterm delivery in labor with intact membranes( n = 8), the term in labor with intact membranes (n = 10), and the term not in labor (control group, n = 12). The expressions of COX-2 in different areas of the fetal membranes were examined by immunohistochemical assay. RESULTS: The immuoreactivity of COX-2 was found in epithelial cells and stromal cells in amniotic and chorion membranes. Expression of COX-2 in different areas of the fetal membranes varied markedly in preterm delivery in labor, term in labor and the control group. Immunohistochemical scores(IH score) of COX-2 in the cervical and corpus fetal membranes in the preterm delivery in labor group were higher than those in the term in labor group and the control group(P < 0.05). There was no difference in COX-2 expression between the cervical and corpus fetal membranes among the three groups (P > 0.05). CONCLUSION: COX-2 may play an important role in the pathogenesis of premature labor. PMID- 15898433 TI - [Proliferation inhibition of human lung adenocarcinoma cell line A549 transfected by RASSF1A gene]. AB - OBJECTIVE: To determine the effects of exogenous RASSF1A gene on the proliferation and expression of P65 and subunit of NF-kappaB, in lung adenocarcinoma cell line A549. METHODS: pcDNA3.0-RASSF1A and pcDNA3. 0 were introduced into A549 cell line by lipofectin transfection, and the A549 cells stably expressing RASSF1A gene were established by G418 selection. The expression of RASSF1A was detected by Western blotting. The cytobiologic characterizations of the positive clone were analyzed by methythiazoletertraolium (MTT) assay and cytometry. The expressing of P65 was analyzed by RT-PCR and Western blotting. RESULTS: A549 cells stably expressing RASSF1A protein were established by lipofection mediated transfection and selected for further study. Compared with the nontransfected and vector transfected cells, the positive clone cells grew more slowly. Flow cytometric data showed that more positive clone cells went into phase G0/G1 and fewer cells went into phase S. The expression of P65 in nuclear protein in positive clone cells was lower than that of the control group while there was no obvious difference between the expression of p65 mRNA and P65 protein in total protein among the 3 groups. CONCLUSION: RASSF1A gene might suppress the proliferation of A549 cells through blocking the activity of P65 protein. PMID- 15898434 TI - [Pharmacokinetics of erythromycin stinoprate capsule]. AB - OBJECTIVE: To determine the pharmacokinetics of erythromycin stinoprate capsules and to provide guidance for clinical research. METHODS: Thirty healthy volunteers (15 men and 15 women) were divided into 3 groups randomly, each including 5 men and 5 women. Single oral doses of 250, 500 and 750 mg were given to each volunteer. The concentrations of erythromycin propionate and erythromycin base in the plasma were determined by HPLC-MS. RESULTS: All 30 volunteers completed the experiment without adverse reactions. Using 3P87 we analyzed the model and calculated the pharmacokinetic parameters. Three dose groups taking high, middle and low dose were all single compartment model. The pharmacokinetic parameters of erythromycin propionate after taking erythromycin stinoprate capsules were as follows: Low dose group: Ka (2.007 +/- 1.281 )/h, tmax ( actual value) (1.9 +/- 0.6) h, Cmax (437.0 +/- 295.0) microg/L, AUC0-14 (trapezoid area) (1840.2 +/- 1476.87) microg x h/L, Ke (0.329 +/- 0.119)/h, T1/2 (2.45 +/- 0.9) h. Middle dose group: Ka (1.451 +/- 0.380)/h, tmax (1.7 +/- 0.3) h, Cmax (923.1 +/- 217.5) microg/L, AUC0-14 (4542.44 +/- 1579.4) microg x h/L,Ke (0.237 +/- 0.057)/h, T1/2 (3.1 +/- 1.1) h; High dose group: Ka (2.076 +/- 1.559)/h, tmax (1.7 +/- 0.3) h, Cmax (1336.5 +/- 366.0) microg/L, AUC0-14 (7481.5 +/- 2496.2) microg x h/L, Ke (0.266 +/- 0.051)/h, T1/2 (2.7 +/- 0.5) h. The pharmacokinetic parameters of erythromycin were as follows: Low dose group: Ka (1.410 +/- 0.626)/h, tmax (1.8 +/- 0.5) h, Cmax (197.5 +/- 227.6) microLg/L, AUC0-14 (766.4 +/- 981.0) microg x h/L, Ke (0.519 +/- 0.240)/ h, T1/2 (1.6 +/- 0.8) h. Middle dose group: Ka (1.900 +/- 1.049)/h, tmax (1.6 +/- 0.2) h,Cmax (488.3 +/- 216.7) microg/L, AUC0-14( 488.3 +/- 216.7) microg/L, Ke (0.329 +/- 0.057)/h, T1/2(2.2 +/- 0.4) h; High dose group: Ka (1.934 +/- 0.794)/h, tmax (1.7 +/- 0.3) h, Cmax (749.3 +/- 387.2) microg/L, AUC0-14(3820.1 +/- 1966.4) microg x h/L, Ke (0.373 +/- 0.174)/h, T1/2( 2.2 +/- 0.7) h. AUC of both erythromycin propionate and erythromycin base was linearly correlated to the doses; T1/2 was not correlated to the doses, so they followed the first order processes. The pharmacokinetic parameters of erythromycin The erythromycin stinoprate propionate and erythromycin base had no gender differences. Conclusion was absorbed as erythromycin propionate. Cmax reached at about 1.6 h. T1/2 of elimination was 2.4-3.1 h. The active component of erythromycin propionate was erythromycin. Cmax of erythromycin is 1.8, T1/2 is 2.4-3.1 h. In the range of oral dose of 250 to 750 mg, both erythromycin propionate and erythromycin base accorded the first order processes. The pharmacokinetic parameters were different with those reported in foreign documents while the gender difference did not exist in Chinese adults. PMID- 15898435 TI - [Proatherogenic effects of immune complexes of human oxLDL in vitro]. AB - OBJECTIVE: To observe the effects of immune complexes (IC) prepared from human oxidized density lipoprotein (oxLDL) antibodies and human oxLDL on the foam cell forming and the macrophage activation, and to further uncover the possible mechanisms of immune complexes contributing to the atherosclerosis occurrence. METHODS: The immune complexes of human oxLDL and purified human oxLDL antibodies were added to culture U937 cells by protocols: polyethylene glycol-precipitated insoluble IC (PEG-IC) and IC immobilized by absorption to red blood cells (RBC IC). With oxLDL as controls and heat-aggregated gamma globulin as an inhibitor of Fc gamma receptor, we measured the cholesterol ester, total cholesterol of the cellular extracts, and quantified the secreted MMP-1 of supernatants from U937 cells. RESULTS: A significant increase of MMP-1 release [(0.769 +/- 0.030) ng/ml vs (0.513 +/- 0.034) ng/ml, P < 0.01] and a higher level of cholesterol ester accumulation [(20.271 +/- 1.668) microg/mg protein vs (17. 226 +/- 1.298 ) microg/mg protein, P < 0.05] in U937 cells incubated with RBC-IC were observed, compared with those incubated with RBC-oxLDL. However, the above quantative difference between the cholesterol ester accumulation induced by oxLDL and insoluble PEG-IC was even more striking, and cholesterol ester accumulation was dosage-dependent. Heat-aggregated gamma globulin (10 mg/ml) as an inhibitor of Fc gamma receptors competitively inhibited cholesterol ester accumulation and decreased PEG-IC stimulating MMP-1 secretion to 71%. CONCLUSION: Immune complexe of ox-LDL can transform macrophages into foam cells and activted macrophages. The immunological function of oxLDL is involved in the process of atherosclerosis occurrence. PMID- 15898436 TI - [Clinical observation and expression of C-reactive protein by LPS-induced A549 cells in patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To investigate the level of mRNA expression of C-reactive proteins (CRP) induced by LPS in human alveolar type epithelial cell line, A549. METHODS: Sputum and plasma specimens were obtained from patients with chronic obstructive pulmonary disease (COPD) diagnosed according to the national criteria. CRP was measured by enzyme-linked immunosorbent assays (ELISA). A549 cells were incubated with lipopolysaccharide (LPS) at different concentrations and for different time. CRP mRNA was extracted from cells and the expression was analyzed by RT-PCR. The CRP in the supernatant was measured by ELISA. RESULTS: The concentration of CRP in the sputum of patients with COPD was significantly higher than that in the plasma (P < 0.05). LPS at different concentrations and for different time induced the expression and secretion of CRP in A549 with a time-dependent increase and the expression of CRP were significantly higher than those of the control (P < 0.05 - 0.01). CONCLUSION: Self-secretion of CRP in the respiratory tract may exist. A549 induced by LPS may express CRP mRNA. PMID- 15898437 TI - [Myocardial protective effects of pretreatment with captopril on ischemia reperfusion myocardium in rabbits]. AB - OBJECTIVE: To investigate the effect of pretreatment with captopril on myocardium ischemia-reperfusion injury in atherosclerotic rabbits. METHODS: Thirty-two New Zealand white rabbits were assigned randomly to the normally feed group, cholesterol-feed (CF) group, and cholesterol food plus captopril group (cap-feed group), which were fed for 10 weeks. We examined the changes in the size of the infarct and changes in the myocardium ultrastructure resulting from coronary ischemia/reperfusion. Levels of endothelin (ET) and nitic oxide (NO) were measured in the different experiment stages. RESULTS: The ET levels significantly increased and the content of NO significantly decreased in the CF group compared with those of the cap-feed group. The ultrastructure of myocardium cell was slightly destroyed and the infarct size was significantly smaller in the cap-feed group than the normally feed rabbits and CF rabbits. CONCLUSION: The long-term captopril treatment can lighten the severity of myocardial injury produced by coronary ischemia/reperfusion. PMID- 15898438 TI - [Modified low-potassium dextran solution in heart-lung transplantation]. AB - OBJECTIVE: To determine the application of modified low-potassium dextran (LPD) solution in heart-lung transplantation. METHODS: We used the modified LPD as the lung flush solution of the donor in the first heart-lung transplantation procedure in Hunan. RESULTS: The patient survived 555 days after the surgery and severe infection, lung disfunction or severe graft rejection never occurred. CONCLUSION: Modified LPD as lung flush solution of the donor during heart-lung transplantation demonstrates an excellent effect of lung protection. PMID- 15898439 TI - [Combining exchange of cerebrospinal fluid with small dose of urokinase injection for subarachnoid hemorrhage]. AB - OBJECTIVE: To explore the curative effect of combining exchange of cerebrospinal fluid with small dose of urokinase injection for the treatment of subarachnoid hemorrhage (SAH). METHODS: One hundred thirty-four SAH patients diagnosed by CT or MRI and lumbar puncture were randomly divided into two groups: 68 patients in the treatment group were given exchange of cerebrospinal fluid and small dose of urokinase injection; 66 patients in the control group were treated with exchange of cerebrospinal fluid. The main complications, the neurological deficit scale and curative effect of the two groups were compared. RESULTS: Complications about cerebral vasospasm and hydrocephalus were fewer than those in the conventional therapeutic group (P < 0.05), and there was no significant difference in the incidence of rebleeding between the two groups (P > 0.05). The neurological deficit scale in the treatment-group was significantly lower than that in conventional therapeutic group (P < 0.01). There was obvious difference the curative effect in the two groups (P < 0.05). CONCLUSION: Combining exchange of cerebrospinal fluid with small dose of urokinase injection is an effective method for treating SAH, and it can improve the survival rate and life quality of SAH patients. PMID- 15898440 TI - [Diagnosis and surgical treatment of 102 cases of ventricular septal defect with patent ductus arteriosus]. AB - OBJECTIVE: To summarize the experience of diagnosis and surgical treatment of ventricular septal defect with patent ductus arteriosus. METHODS: We retrospectively analyzed the clinical data of 102 cases of ventricular septal defect combined with patent ductus arteriosus who underwent surgical treatment. Preoperative ultrasonic cardiogram (UCG) showed ventricular septal defect combined with patent ductus arteriosus in 82 cases and ventricular septal defect in 20 cases. RESULTS: The hospital mortality was 4.9% (5/102). The reasons for death included low cardiac output syndrome (1 case), pulmonary hypertension crisis (2 cases) and respiratory failure (2 cases). In the remaining patients,the perioperative complications included lung infection (7 cases), pulmonary atelectasis (5 cases), hydrothorax (1 case), and pulmonary hypertension crisis (2 cases); and all the 15 patients recovered lastly. The pulmonary hypertension of all living patients decreased to some degree. The therapeutical effectiveness was satisfactory. CONCLUSION: Ventricular septal defect with patent ductus arteriosus is easy to be confused with ventricular septal defect clinically. At the same time,it is diffcult to form a correct diagnosis in some patients by UCG preoperatively. To prevent the occurrence of perfusive lung, it is important to reinforce preoperative diagnosis and exploration during operation. Because pulmonary hypertension in patients with ventricular septal defect with patent ductus arteriosus emerges early and develops quickly, it tends to result in organic pulmonary hypertension which can make patients lose operation chances and influence the long-term therapeutical effect. Surgical operation should be performed as soon as possible. Optimal operative timing and proper perioperative management play important roles in surgical results. PMID- 15898441 TI - [Obesity and diseases in the elderly inpatient]. AB - OBJECTIVE: To explore the obesity distribution in old people and the relation between senile obesity and health. METHODS: First, a questionnaire was designed which included chronic disease history, body mass index (BMI), physiological value, biochemistry index, anti-oxidation index, diagnosis of diseases, etc. Second, the measure and detection methods were unified; and the last, the investigation was made along with daily clinical work by clinicians. RESULTS: We received 391 questionnaires. The overweight rate was 36.1% and the obesity rate was 7.9% . Total anti-oxidation activity in serum (TAS) and superoxide dismutase (SOD) decreased with body mass index (BMI), and the value in the obesity group was the lowest; Malonaldehyde (MDA) of overweight obesity was the largest. The mean blood pressure, blood fat, and blood glucose as well as the prevalence of cardiovascular disease, hyperlipemia, and glycuresis increased with BMI; and the value in the obesity group was the largest. CONCLUSION: The prevalence of the senile obesity was below the average and the senile obesity complications were various and serious, and perhaps related to imbalance of free radical's production and cleanup, so the senile obesity seriously harmed old people's health. PMID- 15898442 TI - [Development of the comprehensive evaluation methods in medicine]. PMID- 15898443 TI - [Development of platelet-associated tissue factors]. PMID- 15898444 TI - [Polymorphism of 13 STR loci of Han population in Xi'an]. PMID- 15898445 TI - [Comparison of clinical effect of sufentanil and fentanyl during heart value replacement surgery]. PMID- 15898446 TI - [Analysis of prognosis and cause of disease of vitreous hemorrhage in elderly patients]. PMID- 15898447 TI - [Relationship of low density lipoprotein cholesterol and insulin-like growth factor binding protein-1 in normal subjects]. PMID- 15898448 TI - [Diagnosis and differential diagnosis of visceral leishmaniasis in Hunan province]. PMID- 15898449 TI - Comparing Social Security reform options. AB - Social Security is widely recognized as the nation's most effective anti-poverty program for the elderly and widow(er)s. It is so popular that it has often been dubbed the "third rail" of American politics ("touch it and you die"). As a result, changes have come slowly. For instance, in spite of years of warning in advance of the cash flow crisis of 1983, Congress waited until the last minute to act--and when it did, the action it took included a combination of tax increases and benefit reductions. By the mid-1990s, then-President Clinton was talking about the long-term financing issues faced by Social Security, but Congress did not act. President Bush has raised the same issues since 2000, and has now taken to the road to convince the nation that action should be taken now to assure the program's long-term solvency. Because Social Security is a sensitive, complicated, and emotional political topic, many concepts have been discussed but few elected officials have been willing to put forth detailed plans for fear of political backlash. The public, quite naturally, wants to know how they will be affected by "reform." In this introductory section, Figure S-1 seeks to provide a simple response to that question by following the method used in the Trustees' report, where earners maintain a constant percentage of the average wage. Take the year closest to when you were born, the earnings closest to your expected earnings this year (2005), and follow across the columns to see how much your annual benefit would be in today's dollars if you start taking benefits at age 65. For an example of a specific individual: Your 30-year-old child (born in 1975) makes a 2005 salary around $16,500. Under current law, your child's initial annual Social Security retirement benefit would be dollar 11,200 in today's dollars. However, given the projected funding shortfall currently facing the program, this promised benefit is not likely to materialize unless some sort of change is made to the program. This analysis compares "Model 2" from the President's 2001 Commission to Strengthen Social Security (which appears to have the principles for an individual account plan favored by the Bush administration) with three basic options: Current-law benefits with taxes raised to cover the shortfall over the 75-year actuarial period, by removing the existing dollar 90,000 annual wage cap and including all workers. Maintain current benefits until the revenue shortfall occurs, when a "cliff" benefit cut is imposed. A gradual reduction in current-law benefits. PMID- 15898451 TI - Facility profile. Design helps rural hospital achieve regional status. PMID- 15898452 TI - In the bag. Using commissioning to improve new building performance. PMID- 15898453 TI - Hear, hear. Assessing and resolving hospital noise issues. PMID- 15898454 TI - Talking technology. Are health care facilities ready for VoIP? PMID- 15898455 TI - Chemical reaction. A look at OSHA's guidance for chemical incident first receivers. PMID- 15898456 TI - Into the future. Key developments likely to affect health facility cleaning. PMID- 15898457 TI - Familial overlap between bipolar disorder and psychotic symptoms in a Canadian cohort. AB - BACKGROUND: Although they were once considered separate nosologic entities, there is current interest in the etiologic overlap between bipolar disorder (BD) and schizophrenia. A critical issue concerns the familial basis of the overlap, specifically, the possibility of a distinct familial subtype of BD with psychotic features. METHODS: We recruited individuals with BD from the community and compared them with a matched group diagnosed with no mental disorder to confirm familial aggregation for BD, schizophrenia, and psychotic symptoms. We then compared BD probands both with and without first-degree relatives with psychotic symptoms on several clinical indicators to determine the specificity of the familial aggregation. RESULTS: As expected, there was evidence for familial aggregation of schizophrenia and psychotic symptoms in families having probands with BD. Familial loading for schizophrenia and psychotic symptoms was especially notable in male relatives of female probands with BD. We found no differences in the clinical profile of probands with BD stratified for familial loading for psychotic symptoms. CONCLUSIONS: Findings from this sample support etiologic theories arguing for a shared but nonspecific genetic etiology for BD and schizophrenia, with psychotic symptoms being a potential key indicator for genetic studies. PMID- 15898458 TI - Long-term medical conditions and major depression: strength of association for specific conditions in the general population. AB - BACKGROUND: The prevalence of major depression (MD) in persons with nonpsychiatric medical conditions is an indicator of clinical need in those groups, an indicator of the feasibility of screening and case-finding efforts, and a source of etiologic hypotheses. This analysis explores the prevalence of MD in the general population in relation to various long-term medical conditions. METHODS: We used a dataset from a large-scale Canadian national health survey, the Canadian Community Health Survey (CCHS). The sample consisted of 115 071 subjects aged 18 years and over, randomly sampled from the Canadian population. The survey interview recorded self-reported diagnoses of various long-term medical conditions and employed a brief predictive interview for MD, the Composite International Diagnostic Interview Short Form for Major Depression. Logistic regression was used to adjust estimates of association for age and sex. RESULTS: The conditions most strongly associated with MD were chronic fatigue syndrome (adjusted odds ratio [AOR] 7.2) and fibromyalgia (AOR 3.4). The conditions least strongly associated were hypertension (AOR 1.2), diabetes, heart disease, and thyroid disease (AOR 1.4 in each case). We found associations with various gastrointestinal, neurologic, and respiratory conditions. CONCLUSIONS: A diverse set of long-term medical conditions are associated with MD, although previous studies might have lacked power to detect some of these associations. The strength of association in prevalence data, however, varies across specific conditions. PMID- 15898459 TI - Challenges facing child psychiatry in Quebec at the dawn of the 21st Century. AB - OBJECTIVES: This study aimed to identify hospital resources by region, determine human resources by type of service and region, and describe how services generally operate in child psychiatry within the province of Quebec. METHODS: Data collection took place from May to October 2001. We sent a semistructured questionnaire to all child psychiatry service heads. We collected human resource data and produced organizational charts based on the responses obtained. These charts were forwarded to each of the participating services for validation. We grouped Quebec's 18 social health regions into 3 categories: central (4 regions with 606 370 youths), adjoining (4 regions with 589 750 youths), and peripheral (10 regions with 368 635 youths). RESULTS: The response rate was 100%. We identified 35 child psychiatry services: 13 in the central regions, 9 in the adjoining regions, and 13 in the peripheral regions. Overall, we identified 177 short-stay beds, 476 places in day or evening hospitals, and 113 places in day or evening centres. Most of these resources were located in the central regions. Quebec had 138.2 full-time equivalent (FTE) child psychiatrists (69.8% in the central regions) and 706 FTE professionals. At March 31, 2001, 4285 youths were waiting for services. CONCLUSIONS: We observed a shortage of child psychiatrists and professionals, regardless of the norm used. Adjoining and peripheral regions should have access to a minimal range of human and hospital resources in child psychiatry. PMID- 15898460 TI - A national survey of gambling problems in Canada. AB - OBJECTIVE: The 1990s saw widespread expansion of new forms of legalized gambling involving video lottery terminals (VLTs) in community settings (that is, in bars and restaurant lounges) and permanent casinos in several Canadian provinces. To date, there has never been a national survey of gambling problems with representative interprovincial data. Using a new survey, we sought to compare prevalence figures across the 10 Canadian provinces. METHOD: Using the Canadian Problem Gambling Index, we investigated the current 12-month prevalence of gambling problems in the Canadian Community Health Survey: Cycle 1.2--Mental Health and Well-Being, in which a random sample of 34,770 community-dwelling respondents aged 15 years and over were interviewed. The response rate was 77%. The data are representative at the provincial level and were compared with the availability of VLTs per 1000 population and with the presence of permanent casinos for each province. RESULTS: Manitoba (2.9%) and Saskatchewan (also 2.9%) had the highest prevalence of gambling problems (specifically, moderate and severe problem levels combined). These 2 provinces had significantly higher levels than the 2 provinces with the lowest prevalence of gambling problems: Quebec (1.7%) and New Brunswick (1.5%). CONCLUSIONS: The 12-month prevalence of gambling problems in Canada was 2.0%, with interprovincial variability. The highest prevalence emerged in areas with high concentrations of VLTs in the community combined with permanent casinos. These findings support earlier predictions that the rapid and prolific expansion of new forms of legalized gambling in many regions of the country would be associated with a considerable public health cost. PMID- 15898461 TI - Psychiatry in the Nazi era. AB - OBJECTIVES: To update Canadian psychiatrists on recent information from newly discovered Berlin archives about the actions of physicians, especially psychiatrists, during the era of National Socialism in Germany and to encourage introspection about the role of the medical profession, its relationship with government, and its vulnerability to manipulation by ideology and economic pressures. METHOD: This is a selective review of the literature on the collaboration of physicians, especially psychiatrists, in the sterilization, experimentation, and annihilation of patients with mental illness before and during World War II. RESULTS: Directed to value the health of the nation over the care of individual patients and convinced that a hierarchy of worth distinguished one person from another, German psychiatrists were enlisted to commit atrocities during the Nazi period. CONCLUSIONS: The values of care and compassion can be eroded; this knowledge demands constant vigilance. PMID- 15898462 TI - A public health strategy to improve the mental health of Canadian children. AB - Mental health problems are the leading health problems that Canadian children currently face after infancy. At any given time, 14% of children aged 4 to 17 years (over 800,000 in Canada) experience mental disorders that cause significant distress and impairment at home, at school, and in the community. Fewer than 25% of these children receive specialized treatment services. Without effective prevention or treatment, childhood problems often lead to distress and impairment throughout adulthood, with significant costs for society. Children's mental health has not received the public policy attention that is warranted by recent epidemiologic data. To address the neglect of children's mental health, a new national strategy is urgently needed. Here, we review the research evidence and suggest the following 4 public policy goals: promote healthy development for all children, prevent mental disorders to reduce the number of children affected, treat mental disorders more effectively to reduce distress and impairment, and monitor outcomes to ensure the effective and efficient use of public resources. Taken together, these goals constitute a public health strategy to improve the mental health of Canadian children. PMID- 15898463 TI - Major depression in patients with borderline personality disorder: a clinical investigation. AB - OBJECTIVE: Borderline personality disorder (BPD) is characterized by a high frequency of comorbidity with major depressive disorder (MDD). This study aimed to compare the clinical characteristics of 2 groups of patients with MDD: those with concomitant BPD and those with other concomitant personality disorders. METHODS: We assessed 119 outpatients, using a semistructured interview for demographic and clinical features, the Structured Clinical Interview for DSM-IV, Hamilton anxiety and depression scales, the Zung Self-Rating Depression Scale (ZSDS), the Social and Occupational Functioning Assessment Scale (SOFAS), the Sheehan Disability Scale, and the Revised Childhood Experiences Questionnaire. We performed a regression analysis, using the number of criteria for BPD as the dependent variable. RESULTS: Severity of BPD was positively related to the ZSDS score, to self-mutilating behaviours, and to the occurrence of mood disorders in first-degree relatives; it was negatively related to the SOFAS score and age at onset of MDD. CONCLUSIONS: Patients with comorbid MDD and BPD present differential characteristics that indicate a more serious and impairing condition with a stronger familial link with mood disorders than is shown by depression patients with other Axis II codiagnoses. PMID- 15898464 TI - How relevant is seizure duration in assessing the effectiveness of electroconvulsive therapy? PMID- 15898465 TI - Lithium and marijuana withdrawal. PMID- 15898466 TI - Risperidone treatment of periodic catatonia. PMID- 15898467 TI - Lamotrigine-induced neutropenia. PMID- 15898468 TI - Galactorrhea with aripiprazole. PMID- 15898469 TI - Effects of rivastigmine in a case of residual schizophrenia. PMID- 15898470 TI - Aripiprazole reduces alcohol use. PMID- 15898471 TI - A functional analytic approach to computer-interactive mathematics. AB - Following a pretest, 11 participants who were naive with regard to various algebraic and trigonometric transformations received an introductory lecture regarding the fundamentals of the rectangular coordinate system. Following the lecture, they took part in a computer-interactive matching-to-sample procedure in which they received training on particular formula-to-formula and formula-to graph relations as these formulas pertain to reflections and vertical and horizontal shifts. In training A-B, standard formulas served as samples and factored formulas served as comparisons. In training B-C, factored formulas served as samples and graphs served as comparisons. Subsequently, the program assessed for mutually entailed B-A and C-B relations as well as combinatorially entailed C-A and A-C relations. After all participants demonstrated mutual entailment and combinatorial entailment, we employed a test of novel relations to assess 40 different and complex variations of the original training formulas and their respective graphs. Six of 10 participants who completed training demonstrated perfect or near-perfect performance in identifying novel formula-to graph relations. Three of the 4 participants who made more than three incorrect responses during the assessment of novel relations showed some commonality among their error patterns. Derived transfer of stimulus control using mathematical relations is discussed. PMID- 15898472 TI - Behavioral assessment of impulsivity: a comparison of children with and without attention deficit hyperactivity disorder. AB - We conducted a brief computer-based assessment involving choices of concurrently presented arithmetic problems associated with competing reinforcer dimensions to assess impulsivity (choices controlled primarily by reinforcer immediacy) as well as the relative influence of other dimensions (reinforcer rate, quality, and response effort), with 58 children. Results were compared for children with attention deficit hyperactivity disorder (ADHD) who were and were not receiving medication, and with typically developing children without ADHD. Within-subject and between-groups analyses of the ordinal influence of each of the reinforcer dimensions were conducted using both time- and response-allocation measures. In general, the choices of children with ADHD were most influenced by reinforcer immediacy and quality and least by rate and effort, suggesting impulsivity. The choices of children in the non-ADHD group were most influenced by reinforcer quality, and the influence of immediacy relative to the other dimensions was not statistically significant. Results are discussed with respect to the implications for assessment and treatment of ADHD. PMID- 15898473 TI - Using food redistribution to reduce packing in children with severe food refusal. AB - Positive- and negative-reinforcement-based procedures typically have targeted acceptance for children with severe food refusal; however, these procedures do not always result in successful swallowing. Once acceptance is achieved, some children expel the food repeatedly or pack (hold or pocket) it in their mouths for extended periods of time. This study evaluated the effects of using food redistribution with a bristled massaging toothbrush to reduce packing and increase consumption in 4 children with severe feeding disorders. Packing was reduced for all children. In addition, latency to clean mouth (the duration of time from acceptance to food no longer being present in the child's mouth in the absence of expulsion) for 2 children decreased when the food-redistribution procedure was used. Results are discussed in terms of the potential operant functions of the food-redistribution procedure. PMID- 15898474 TI - On the effectiveness of and preference for punishment and extinction components of function-based interventions. AB - The current study describes an assessment sequence that may be used to identify individualized, effective, and preferred interventions for severe problem behavior in lieu of relying on a restricted set of treatment options that are assumed to be in the best interest of consumers. The relative effectiveness of functional communication training (FCT) with and without a punishment component was evaluated with 2 children for whom functional analyses demonstrated behavioral maintenance via social positive reinforcement. The results showed that FCT plus punishment was more effective than FCT in reducing problem behavior. Subsequently, participants' relative preference for each treatment was evaluated in a concurrent-chains arrangement, and both participants demonstrated a dear preference for FCT with punishment. These findings suggest that the treatment selection process may be guided by person-centered and evidence-based values. PMID- 15898475 TI - Evaluation of behavioral skills training for teaching abduction-prevention skills to young children. AB - This study examined the effectiveness of individual behavioral skills training in conjunction with in situ training in teaching 13 preschool children abduction prevention skills. Children's performance was measured during baseline, training, and at 2-week, 1-month, and 3-month follow-ups using in situ assessments in which abduction prevention skills were measured in naturalistic settings. Results revealed that all the children learned the skills and all the children available at the 2-week and 1-month follow-ups maintained the skills at criterion level. All but 3 children's criterion-level performances were maintained at the 3-month follow-up as well. PMID- 15898476 TI - Effects of the contingency for homework submission on homework submission and quiz performance in a college course. AB - Effects of the contingency for submission of homework assignments on the probability of assignment submission and on quiz grades were assessed in an undergraduate psychology course. Under an alternating treatments design, each student was assigned to a points condition for 5 of 10 quiz-related homework assignments corresponding to textbook chapters. Points were available for homework submission under this condition; points were not available under the no points condition. The group-mean percentage of homework assignments submitted and quiz grades were higher for all chapters under the points condition than in the no-points condition. These findings, which were replicated in Experiment 2, demonstrate that homework submission was not maintained when the only consequences were instructor-provided feedback and expectation of improved quiz performance. PMID- 15898477 TI - A systematic evaluation of food textures to decrease packing and increase oral intake in children with pediatric feeding disorders. AB - This study examined packing (pocketing or holding accepted food in the mouth) in 3 children who were failing to thrive or had inadequate weight gain due to insufficient caloric intake. The results of an analysis of texture indicated that total grams consumed were higher when lower textured foods were presented than when higher textured foods were presented. The gram intake was related directly to levels of packing. That is, high levels of packing were associated with higher textured foods and low gram intake, and low levels of packing were associated with lower textured foods and high gram intake. All participants gained weight when texture of foods was decreased. Packing remained low during follow-up for 2 participants even when the texture of food was increased gradually over time. These data are discussed in relation to avoidance, response effort, and skill deficit. PMID- 15898478 TI - Establishing derived requesting skills in adults with severe developmental disabilities. AB - This project examined whether a history of reinforced relational responding would result in derived requesting skills in 3 adults with disabilities. Participants were first taught to request preferred items using pictures; they were then taught conditional discriminations between pictures and their dictated names and between dictated names and their corresponding text. Finally, requests for preferred items using corresponding text were evaluated. All 3 participants demonstrated derived requesting skills. PMID- 15898479 TI - The effects of establishing operations on preferences for tangible items. AB - Researchers have demonstrated that both deprivation and satiation can affect the outcome of preference assessments for food. In the current study, paired-stimulus preference assessments for tangible items were conducted under three conditions: control, deprivation, and satiation. Three persons with developmental disabilities and 3 typically developing preschool children served as participants. The results demonstrated that deprivation and satiation influenced the outcome of preference assessments of leisure items or toys. PMID- 15898480 TI - Use of a lag differential reinforcement contingency to increase varied selections of classroom activities. AB - The present study evaluated the effects of a lag differential reinforcement contingency on 2 students' activity selections using reversal designs. Results showed that the lag contingency was responsible for promoting increased novel selections, engagement in diverse activities, and greater progress with respect to programmed academic activities. PMID- 15898481 TI - Using task clarification, graphic feedback, and verbal feedback to increase closing-task completion in a privately owned restaurant. AB - An informant functional assessment was used to evaluate closing-task completion by servers and dishwashers at a restaurant. Based on the functional assessment results, an intervention consisting of task clarification, posted graphic feedback, and verbal feedback was implemented and evaluated with a multiple baseline design across two groups of employees. Results showed an increase of 15% and 38% in task completion for the two groups. PMID- 15898482 TI - Superimposition and withholding of edible consequences as treatment for automatically reinforced stereotypy. AB - The delivery and subsequent withholding of tangible consequences has been previously investigated as an intervention for stereotypic behavior. The current investigation sought to extend previous research by evaluating its effectiveness and durability as treatment for stereotypy of 2 children who had been diagnosed with autism. Nonsocial functions for stereotypic behavior were identified via functional analysis. Edible items were then delivered contingent on stereotypy and were withheld in a subsequent condition. When the superimposition procedure failed to reduce stereotypy, environmental enrichment was implemented and was found to reduce the stereotypy of both participants. PMID- 15898483 TI - The effects of methylphenidate on a functional analysis of disruptive behavior: a replication and extension. AB - In the present investigation, a functional analysis of the disruptive behavior of a 18-year-old man who had been diagnosed with attention deficit hyperactivity disorder and moderate mental retardation was conducted, both when he was taking methylphenidate and when he was not taking the medication. The results of this functional analysis demonstrated that the participant's disruptive behaviors were reinforced by access to attention only when he was not taking methylphenidate. PMID- 15898484 TI - Using brief experimental assessment of reading interventions for identification and treatment of a vocal habit. AB - An 11-year-old boy presented in an outpatient clinic with a vocal habit that occurred during reading and conversation. A brief reading assessment was conducted to determine an effective intervention to decrease the habit. A modified version of the word error-correction procedure resulted in positive changes and was implemented by his mother during home reading practices. Significant decreases in the rate of vocal habit were observed during home reading probes, generalization probes, and follow-up. PMID- 15898485 TI - Impossible dreams. PMID- 15898486 TI - The humanitarian imperative in disaster management--a memorial tribute to Professor Peter Safar. PMID- 15898487 TI - Impact of 2003 power outages on public health and emergency response. AB - INTRODUCTION: In 2003, a major power outage occurred in the midwest and northeast United States affecting some 50 million people. The power outages affected multiple systems in state and local municipalities and, in turn, affected public health. METHODS: Semi-structured interviews were conducted using open-ended questionnaires, with a convenience sample of state- and locally selected subject matter experts from Ohio, Michigan, and New York. Respondents were interviewed in groups representing one of five areas of interest, including: (1) emergency preparedness; (2) hospital and emergency medical services; (3) municipal environmental systems; (4) public health surveillance and epidemiology; and (5) psychosocial and behavioral issues. The reported positive and negative impacts of the power outage on public health, medical services, and emergency preparedness and response were documented. Responses were categorized into common themes and recommendations were formulated. RESULTS: The amount of time that the respondents' locations were without power ranged from <1 hour to 52 hours. Many common themes emerged from the different locations, including communications failures, alternate power source problems, manpower and training issues, and psychosocial concerns. There was minimal morbidity and mortality reported that could be attributed to the event. CONCLUSION: Power outages negatively impacted multiple municipal infra-structures, and affected medical services, emergency response, and public health efforts. Previous federal funding positively impacted public health and emergency response capabilities. Recommendations were made based upon the common themes identified by the respondents. Recommendations may assist state and local health departments, medical service providers, and emergency responders in planning for future power outage problems. PMID- 15898488 TI - Mass medical repatriation of injured civilians after terrorist attack in Mombassa, Kenya: medical needs, resources used, and lessons learned. AB - INTRODUCTION: On 28 November 2002, three suicide bombers crashed their car into a hotel in Mombassa, Kenya; 12 people were killed, including three Israelis, and 80 were wounded (22 of whom were Israeli). The Israeli Defense Force Airborne Medical Evacuation Flight Teams participated in a repatriation mission to bring the wounded home. OBJECTIVES: The objectives of this study are to outline the distinctive aspects of this mission, as well as to share the experiences and lessons learned. METHODS: Israeli Army debriefing reports were used to study the composition of the crew, medical equipment taken, injury distribution, mode of operation, and mission schedule. RESULTS: A total of six fixed-wing aircraft were used--two Boeing 707s and four Hercules C-130s--with a total of 54 medical team members on board. A total of 260 Israelis were repatriated, 22 of whom were wounded, and three were dead. Of the casualties, 14 were conveyed sitting, and eight supine. The time from the first landing in Kenya to the evacuation of the last supine patient was 5.5 hours. Nurses, as well as social workers, played a central role in the mission. A forward team, including five doctors, was used for the initial organization and for gathering information on the medical status of the casualties. CONCLUSIONS: There was redundancy in the medical crew and medical equipment sent. The need for improved infrastructure on the medical aircraft was stressed. Based on this experience, a new mode for operation for similar missions in the future was formulated. PMID- 15898489 TI - A multidisciplinary field hospital as a substitute for medical hospital care in the aftermath of an earthquake: the experience of the Israeli Defense Forces Field Hospital in Duzce, Turkey, 1999. AB - The damage created by an earthquake can overwhelm local health services, and damage to clinics and hospitals can render them useless. After an earthquake, even undamaged medical facilities cannot be used for a period of time if there is a risk of aftershocks and collapse. In such a situation, there may be calls for international health teams--but what constitutes the optimal medical aid a few days after the event? Does a military field hospital fill the "gap" in the local healthcare system? On 12 November 1999, a 7.2 magnitude earthquake struck Duzce, Turkey. All of the medical activities of the responding Israeli Defense Forces (IDF) mission team field hospital in Duzce, Turkey were recorded and evaluated. A total of 2,230 patient contacts occurred at the field hospital during the nine days it operated. Most of the patients who presented (90%) had non-traumatic medical, pediatric, or gynecological problems unrelated to the earthquake. The IDF hospital offered medical care provided by specialists, hospitalization, and surgical abilities, which Duzce's hospitals could not offer until two weeks after the earthquake. These results strengthen the importance of a multidisciplinary, versatile, field hospital as an aid to an earthquake-affected population during the first few weeks after an earthquake. PMID- 15898490 TI - Peer evaluation of the professional behaviors of emergency medical technicians. AB - INTRODUCTION: Professional behavior is one of the cornerstones of effective emergency medical services (EMS) practice and is a required part of the National Standard Curricula for advanced levels of EMS education. However, peer rating of emergency medical technicians with respect to the 11 categories of professional behavior never has been quantified. This study uses a peer evaluation methodology to assess the affective competencies of practicing EMS providers. METHODS: A professional behavior evaluation form was included as part of a survey that was sent to 2,443 randomly selected, nationally registered emergency medical technicians (EMTs). Participants were asked to rate the EMT partner with whom they worked most closely in the past year using 11 different categories of professional behavior using a Likert scale. RESULTS: One thousand, five hundred, ten (61.8%) surveys were returned and analyzed. Both nationally registered EMTs at the Basic and Paramedic levels rated their partners with respect to 11 categories of professional behavior. The overall average score was 0.68 on a 0-1 scale, with one being the highest. The rating of each of the categories was: (1) integrity (0.77); (2) appearance/personal hygiene (0.74); (3) patient advocacy (0.73); (4) empathy (0.72); (5) self-confidence (0.70); (6) careful delivery of service (0.70); (7) respect (0.65); (8) communication skills (0.64); (9) time management skills (0.63); (10) teamwork/diplomacy skills (0.62); and (11) self motivation (0.61). Overall, the NREMT-Paramedics rated their partners significantly lower than did the NREMT-Basics (p = 0.0156) and experienced EMT Basics rated their partners significantly lower than did the newer EMT-Basics (p = 0.0002). Those EMTs who indicated high satisfaction with their current EMS assignment rated their partner more highly on professional behaviors than did those EMTs who were not as satisfied. CONCLUSION: Overall, EMTs peer evaluation of professional behavior was "good." The behaviors most highly rated were integrity and appearance/personal hygiene. The behaviors rated lowest were self motivation and team work/diplomacy. It appears that paramedics are more critical of their colleagues than are EMT-Basics, that experienced EMT-Basics are harsher critics than are newer EMT-Basics, and that there is a relationship between job satisfaction and peer evaluation. PMID- 15898491 TI - Occupational stress among Japanese emergency medical technicians: Hyogo Prefecture. AB - INTRODUCTION: As prehospital care became emphasized in emergency medical services in Japan, qualification as a "paramedic" was established in 1991 as a requirement for national qualification as a emergency medical technician (EMT). With recent increases in emergency transportation, the responsibilities of paramedics have become more complex and demand a higher level of competency; however, no method of evaluating occupational stress among Japanese EMTs currently exists. METHODS: A questionnaire survey of the working conditions and health of 2,017 EMTs in Hyogo Prefecture was conducted. To analyze stress levels among these EMTs, the survey was divided into two categories: (1) physical stress; and (2) mental stress. RESULTS: The number of responses was 1,551 (76.9%) and the average age of the respondents was 35.4 years. The lower back, neck, and shoulders were most frequently subjected to physical stress, which was related to the daily operations as an EMT. Mental stress was reported more frequently by those who were older or qualified paramedics. DISCUSSION: The high frequency of lower back pain suggests the need for improvement in the work environment and periodic education. CONCLUSIONS: Although job satisfaction among paramedics was high, they were exposed to greater mental stress. Therefore, systematic management of stress must be developed and established. PMID- 15898492 TI - Prehospital management of earthquake casualties buried under rubble. AB - Earthquakes continue to exact a heavy toll on life, injury, and loss of property. Survival of casualties extricated from under the rubble depends upon early medical interventions by emergency teams on site. The objective of this paper is to review the pertinent literature and to analyze the information as a practical guideline for the medical management of casualties accidentally buried alive. PMID- 15898493 TI - Fire safety knowledge and practices among residents of an assisted living facility. AB - INTRODUCTION: Assisted living facilities (ALFs) pose unique fire risks to the elderly that may be linked to specific fire safety (FS) practices. OBJECTIVE: To evaluate self-reported FS practices among ALF residents. METHODS: All residents of a small ALF were surveyed regarding actual and hypothetical FS behaviors, self perceived fire risk, and FS preparedness. RESULTS: Fifty-eight ALF residents completed the survey. Thirty-three (58%) individuals reported one or more disabilities. Seven (12%) residents ignored the fire alarm and 21 (35%) could not hear it clearly. Sixteen (28%) residents would attempt to locate the source of a fire rather than escape from the building. Only 24 (42%) residents were familiar with the building fire plan. Twenty-three (40%) people surveyed believed that they were not at risk of fire in the study facility. CONCLUSION: Residents of an ALF may be at increased fire injury risk due to their FS practices and disabilities. PMID- 15898494 TI - Phytohormones mediate volatile emissions during the interaction of compatible and incompatible pathogens: the role of ethylene in Pseudomonas syringae infected tobacco. AB - Interactions between the phytohormones ethylene, salicylic acid (SA), and jasmonic acid (JA) are thought to regulate the specificity of induced plant defenses against microbial pathogens and herbivores. However, the nature of these interactions leading to induced plant volatile emissions during pathogen infection is unclear. We previously demonstrated that a complex volatile blend including (E)-beta-ocimene, methyl salicylate (MeSA), and numerous sesquiterpenes was released by tobacco plants, Nicotiana tabacum K326, infected with an avirulent/incompatible strain of Pseudomonas syringae pv. tomato (Pst DC3000). In contrast, a volatile blend, mainly consisting of MeSA and two unidentified sesquiterpenes, was released by plants infected with P. syringae pv. tabaci (Pstb) in a virulent/compatible interaction. In this study, we examined the interaction of multiple pathogen stresses, phytohormone signaling, and induced volatile emissions in tobacco. Combined pathogen infection involved the inoculation of one leaf with Pst DC 3000 and of a second leaf, from the same plant, with Pstb. Combined infection reduced emissions of ocimene and MeSA compared to plants infected with Pst DC 3000 alone, but with no significant changes in total sesquiterpene emissions. In the compatible interaction, Pstb elicited a large ethylene burst with a peak emission occurring 3 days after inoculation. In contrast, the incompatible interaction involving Pst DC3000 displayed no such ethylene induction. Pstb-induced ethylene production was not significantly altered by Pst DC3000 in the combined infection. We postulated that Pstb-induced ethylene production may play a regulatory role in altering the typical volatile emission in tobacco in response to Pst DC3000 infection. To clarify the role of ethylene, we dynamically applied ethylene to the headspace of tobacco plants following infection with Pst DC3000. Consistent with Pstb-induced ethylene, exogenous ethylene reduced both ocimene and MeSA emissions, and selectively altered the ratios and amounts of induced sesquiterpene emissions. Our findings suggest that ethylene can regulate the magnitude and blend of induced volatile emissions during pathogen infection. PMID- 15898495 TI - Variation in plant volatiles and attraction of the parasitoid Diadegma semiclausum (Hellen). AB - Differences in allelochemistry of plants may influence their ability to attract parasitoids. We studied responses of Diadegma semiclausum (Hellen), a parasitoid of the diamondback moth (Plutella xylostella L.), to inter- and intraspecific variation in odor blends of crucifers and a non-crucifer species. Uninfested Brussels sprout (Brassica oleracea L. gemmifera), white mustard (Sinapis alba L.), a feral Brassica oleracea, and malting barley (Hordeum vulgare L.) were compared for their attractivity to D. semiclausum in a Y-tube bioassay. Odors from all plants were more attractive to the parasitoid than clean air. However, tested against each other, parasitoids preferred the volatile blend from the three cruciferous species over that of malting barley. Wasps also discriminated between uninfested crucifers: mustard was as attractive as feral B. oleracea, and both were more attractive than Brussels sprout. Attractivity of uninfested plants was compared with that of plants infested by larvae of the host P. xylostella. Host-infested mustard and Brussels sprout were more attractive than uninfested conspecifics. Interestingly, the volatile blends of uninfested white mustard and infested Brussels sprout were equally attractive. We also compared the volatile composition of different plant sources by collecting headspace samples and analysing them with GC-MS. Similarities of volatile profiles were determined by hierarchic clustering and non-metric scaling based on the Horn-index. Due to the absence of several compounds in its blend, the volatile profile of barley showed dissimilarities from blends of crucifers. The odor profile of white mustard was distinctly different from the two Brassicaceae. Feral Brassica oleracea odor profile was different from infested Brussels sprout, but showed overlap with uninfested Brussels sprout. Odor blends from infested and uninfested Brussels sprout were similar, and mainly quantitative differences were found. D. semiclausum appears to discriminate based on subtle differences in volatile composition of odor blends from infested and uninfested plants. PMID- 15898496 TI - Further field evaluation of synthetic herbivore-induced plant volatiles as attractants for beneficial insects. AB - Fifteen synthetic herbivore-induced plant volatiles (HIPVs) were field-tested for attractivity to beneficial insects in two experiments conducted in an open field and a hop yard in Washington State. Eleven insect species or families showed significant attraction to 13 HIPVs. The ladybeetle, Stethorus punctum picipes, was attracted to sticky traps baited with methyl salicylate (MeSA), cis-3-hexen-1 ol (He), and benzaldehyde (Be). The minute pirate bug, Orius tristicolor, was attracted to traps baited with MeSA, He, Be, and octyl aldehyde (Oa), and the bigeyed bug, Geocoris pallens, responded to MeSA, indole, and trans-2-hexen-1-al. The mymarid wasp, Anagrus daanei, was attracted to He, Oa, and farnesene. The chloropid fly, Thaumatomyia glabra, was highly attracted to methyl anthranilate. Insect families responding to HIPVs included Syrphidae (MeSA, He), Braconidae ((Z)-3-hexenyl acetate, He, cis-jasmone (J), methyl jasmonate (MeJA), methyl anthranilate (MeA)), Empididae (MeSA), Sarcophagidae (MeSA, Be, J, nonanal and geraniol), Tachinidae (Be), and Agromyzidae (MeSA). Micro-Hymenoptera (primarily parasitic wasp families) were attracted to MeSA, He, and indole. These results are discussed with respect to known properties and bioactivity of the tested HIPVs and to their potential as tools for recruiting natural enemies into agroecosystems. PMID- 15898497 TI - Synomone or kairomone?--Bulbophyllum apertum flower releases raspberry ketone to attract Bactrocera fruit flies. AB - Bulbophyllum apertum flower (Orchidaceae) releases raspberry ketone (RK) in its fragrance, which attracts males of several fruit fly species belonging to the genus Bactrocera. Besides RK as a major component, the flower contains smaller amounts of 4-(4-hydroxylphenyl)-2-butanol, plus two minor volatile components, veratryl alcohol and vanillyl alcohol. Within the flower, the lip (labellum) had the highest concentration of RK with much smaller quantities present in petals; other flower parts had no detectable RK. Male fruit flies attracted to the flower belong to RK-sensitive species--such as Bactrocera albistragata, B. caudatus, B. cucurbitae (melon fly), and B. tau. Removal and attachment of the pollinarium to a fly's thoracic dorsum occurred when a male of B. albistragata was toppled into the floral column cavity, due to an imbalance caused by it shifting its body weight while feeding on the see-saw lip, and then freeing itself after being momentarily trapped between the lip and column. During this process, the stiff hamulus (the pollinia stalk protruding prominently towards the lip) acted as a crowbar when it was brushed downwards by the toppled fly and lifted the pollinia out of the anther. If the fly was big or long for the small triangular lip, it would not be toppled into the column cavity and would just walk across the column, during which time the pollinarium could be accidentally removed by the fly's leg, resulting in a failed transport of the pollinarium. This suggests an unstable situation, where the orchid relies only on a particular pollinator species in the complex ecosystem where many RK-sensitive species inhabit. Wild males of B. caudatus (most common visitors) captured on Bulbophyllum apertum flowers were found to sequester RK in their bodies as a potential pheromonal and allomonal ingredient. Thus, RK can act either as a floral synomone (pollinarium transported) or kairomone (accidental removal of pollinarium leading to total pollen wastage), depending on the body size of the male fruit flies visiting the flowers. PMID- 15898498 TI - Hydroquinone is not a phagostimulant for the Formosan subterranean termite. AB - It has been suggested that hydroquinone found in the labial glands of a number of termite species acts as a primary phagostimulating factor. We tested hydroquinone as a phagostimulant using workers from three colonies of the Formosan subterranean termite, Coptotermes formosanus, under both laboratory and field conditions. Hydroquinone at concentrations ranging from ca. 0.002-20.0 ng/cm2 did not increase visitation by C. formosanus workers to treated over control filter papers, and was actually repellent at a 20 ng/cm2 dose. No phagostimulant response to hydroquinone was observed in two colonies. In the third, there was a significant increase in feeding on filter paper treated with a 2 ng/cm2 dose, but was significantly lower at a 20 ng/cm2 dose. Furthermore, sand treated with a gradient of hydroquinone, did not evoke increased tunneling activity compared with controls. GC-MS analysis of C. formosanus workers indicated that hydroquinone was present at an average of 41 pg/worker. It was also determined that within one week about 11% hydroquinone in aqueous solution oxidized to 1,4 benzoquinone. Our findings indicate that hydroquinone alone does not act as a phagostimulant but instead may act as a repellent at higher concentrations. The attractant/arrestant of the Formosan termite may have multiple components of which hydroquinone, at low doses, could be one. PMID- 15898499 TI - Inheritance of resistance to mammalian herbivores and of plant defensive chemistry in a Eucalyptus species. AB - Hybridization in plants provides an opportunity to investigate the patterns of inheritance of hybrid resistance to herbivores, and of the plant mechanisms conferring this resistance such as plant secondary metabolites. We investigated how inter-race differences in resistance of Eucalyptus globulus to a generalist mammalian herbivore, Trichosurus vulpecula, are inherited in their F1 hybrids. We assessed browsing damage of three-year-old trees in a common environment field trial on four hybrid types of known progeny. The progency were artificial intra race crosses and reciprocal inter-race F1 hybrids of two geographically distinct populations (races) of E. globulus; north-eastern Tasmania and south-eastern Tasmania. Populations of trees from north-eastern Tasmania are relatively susceptible to browsing by T. vulpecula, while populations from south-eastern Tasmania are more resistant. We assessed the preferences of these trees in a series of paired feeding trials with captive animals to test the field trial results and also investigated the patterns of inheritance of plant secondary metabolites. Our results demonstrated that the phenotypic expression of resistance of the inter-race F1 hybrids supported the additive pattern of inheritance, as these hybrids were intermediate in resistance compared to the pure parental hybrids. The expression of plant secondary metabolites in the F1 hybrids varied among groups of individual compounds. The most common pattern supported was dominance towards one of the parental types. Together, condensed tannins and essential oils appeared to explain the observed patterns of resistance among the four hybrid types. While both chemical groups were inherited in a dominant manner in the inter-race F1 hybrids, the direction of dominance was opposite. Their combined concentration, however, was inherited in an additive manner, consistent with the phenotypic differences in browsing. PMID- 15898500 TI - Effects of tree phytochemistry on the interactions among endophloedic fungi associated with the southern pine beetle. AB - We examined the interaction between host trees and fungi associated with a tree killing bark beetle, Dendroctonus frontalis. We evaluated (1) the response of four Pinus species to fungal invasion and (2) the effects of plant secondary metabolites on primary growth of and secondary colonization of three consistent fungal associates. Two of these fungi, Entomocorticium sp. A and Ophiostoma ranaculosum, are obligate mutualists with D. frontalis, and the third associate is a blue-staining fungus, O. minus, that is commonly introduced by beetles and phoretic mites. O. minus negatively affects beetle larvae and in high abundance can impact D. frontalis population dynamics. Size of lesions formed and quantity of secondary metabolites produced in response to fungal inoculations varied significantly among Pinus species. However, monoterpene composition within infected tissue did not significantly vary across treatments. While all eight tested metabolites negatively affected the growth rate of O. minus, only 4 allylanisole, p-cymene, and terpinene reduced the growth of the mycangial fungi. Surprisingly, growth rates of mycangial fungi increased in the presence of several secondary metabolite volatiles. O. minus out-competed both mycangial fungi, but the presence of secondary metabolites altered the outcome slightly. O. ranaculosum out-performed E. sp. A in the presence of dominant conifer monoterpenes, such as alpha- and beta-pinene. Volatiles from the mycangial fungi, particularly E. sp. A, had a negative effect on O. minus growth. In general, phloem phytochemistry of particular Pinus species appeared to alter the relative growth and competitiveness of mutualistic and non-mutualistic fungi associated with D. frontalis. The outcome of interactions among these fungi likely has important consequences for the population dynamics of D. frontalis. PMID- 15898501 TI - Polyamine profiles of healthy and parasite-infected Vaccinium myrtillus plants under nitrogen enrichment. AB - Addition of nitrogen (N) to the field layer of boreal forests has been shown to increase the occurrence of the parasitic fungus Valdensia heterodoxa on Vaccinium myrtillus plants. We investigated whether N addition to soil alters the levels of polyamines in V. myrtillus shoots, and discuss here whether such changes could promote the spread of the parasitic fungus on V myrtillus. Using HPLC, we analyzed the concentrations of free and conjugated polyamines in healthy and naturally V. heterodoxa-infected V. myrtillus plants, which had received a moderate or high dose of N fertilizer, or no additional N. Fertilization with N increased the concentrations of free diamines (putrescine and diaminopropane), but had no significant effect on conjugated amines. Thus, N-induced changes in the constitutive levels of soluble conjugated amines do not seem to explain the increased parasite susceptibility of V. myrtillus under N enrichment. Generally, the concentrations of free diamines and insoluble conjugated putrescine were higher in diseased than in healthy shoots, suggesting parasite-induced accumulation of diamines. Free spermine seemed to accumulate in unfertilized, diseased plants, but in fertilized plants this induction was dampened, suggesting that N-induced alterations in spermine metabolism may promote the spread of parasites on V. myrtillus under N-enrichment. PMID- 15898502 TI - The effects of humic substances on Pinus callus are reversed by 2,4 dichlorophenoxyacetic acid. AB - The reversal of humic matter-induced inhibition of callus growth and metabolism by 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in Pinus laricio. Two forest humic fractions (relative molecular mass (Mr) > 3500), derived from soil under Fagus sylvatica (Fs) and Abies alba (Aa) plantation, were used. Pinus laricio callus was grown for a subculture period (4 weeks) on Basal Murashige and Skoog (MS) medium plus forest humic matters (Fs or Aa), at a concentration of 1 mg C/l, and then was transferred, for an additional four weeks, to a MS medium culture without humic matter, but with different hormones: indole-3-acetic acid (IAA, 2 mg/l) or 2,4-dichlorophenoxyacetic acid (2,4-D, 0.5 mg/l) and/or 6 benzylaminopurine (BAP, 0.25 mg/l). Growth of calluse, glucose, fructose, and sucrose contents, and activities of soluble and bound invertases, glucokinase, phosphoglucose isomerase, aldolase, and pyruvate kinase were monitored. The results show a negative effect of humic fractions on callus growth, due to decreased utilization of glucose and fructose, and decreased activities of glycolytic enzymes. The effects are reversible. Substitution of humic fractions with 2,4-D+BAP or 2,4-D is followed by an increase of glycolytic enzyme activities and, consequently, by the utilization of glucose and fructose that induces a restart of growth. In contrast, the inhibitory effects of humic fractions persist when they are substituted with BAP alone, indicating that only the auxin 2,4-D is capable of reversing the negative effects. A possible competitive action on the auxin-binding site between 2,4-D and the chemical structures in the forest humic fractions is suggested. PMID- 15898503 TI - Plant growth inhibition by cis-cinnamoyl glucosides and cis-cinnamic acid. AB - Spiraea thunbergii Sieb. contains 1-O-cis-cinnamoyl-beta-D-glucopyranose (CG) and 6-O-(4'-hydroxy-2'-methylene-butyroyl)-1-O-cis-cinnamoyl-beta-D-glucopyranose (BCG) as major plant growth inhibiting constituents. In the present study, we determined the inhibitory activity of CG and BCG on root elongation of germinated seedlings of lettuce (Lactuca sativa), pigweed (Amaranthus retroflexus), red clover (Trifolium pratense), timothy (Phleum pratense), and bok choy (Brassica rapa var chinensis) in comparison with that of two well-known growth inhibitors, 2,4-dichlorophenoxyacetic acid (2,4-D) and (+)-2-cis-4-trans-abscisic acid (cis ABA), as well as two related chemicals of CG and BCG, cis-cinnamic acid (cis-CA) and trans-cinnamic acid (trans-CA). The EC50 values for CG and BCG on lettuce were roughly one-half to one-quarter of the value for cis-ABA. cis-Cinnamic acid, which is a component of CG and BCG, possessed almost the same inhibitory activity of CG and BCG, suggesting that the essential chemical structure responsible for the inhibitory activity of CG and BCG is cis-CA. The cis-stereochemistry of the methylene moiety is apparently needed for high inhibitory activity, as trans-CA had an EC50 value roughly 100 times that of CG, BCG, and cis-CA. Growth inhibition by CG, BCG, and cis-CA was influenced by the nature of the soil in the growing medium: alluvial soil preserved the bioactivity, whereas volcanic ash and calcareous soils inhibited bioactivity. These findings indicate a potential role of cis-CA and its glucosides as allelochemicals for use as plant growth regulators in agricultural fields. PMID- 15898504 TI - (Z,Z)-6,9-Heneicosadien-11-one: major sex pheromone component of painted apple moth, Teia anartoides. AB - (Z,Z)-6,9-Heneicosadien-11-one (Z6Z9-11-one-21Hy) was identified as the major sex pheromone component of the painted apple moth (PAM), Teia anartoides (Lepidoptera: Lymantriidae), on the basis of (1) comparative gas chromatographic electroantennographic detection (GC-EAD) analyses, GC-mass spectrometry (MS), high-performance liquid chromatography (HPLC)-MS, and HPLC-UV/visible spectroscopy of pheromone gland extracts and authentic standards; (2) GC-EAD analyses of effluvia of calling females; and (3) wind tunnel and field trapping experiments with a synthetic standard. In field experiments in Australia, synthetic Z6Z9-11-one-21Hy as a single component attracted male moths. Wind tunnel experiments suggested that a 4-component blend consisting of Z6Z9-11-one 21Hy, (6Z,9R,10S)-cis-9,10-epoxy-heneicosene (Z6-9R10S-epo-21 Hy), (E,E)-7,9 heneicosadien-6, 11-dione (E7E9-6,11-dione-21Hy), and 6-hydroxy-(E,E)-7,9 heneicosadien-11-one (E7E9-6-ol-11-one-21Hy) (all present in pheromone gland extracts) might induce more males to orient toward, approach, and contact the source than did Z6Z9-11-one-21Hy as a single component. Additional experiments are needed to determine conclusively whether or not Z6-9R10S-epo-21Hy, E7E9-6,11 dione-21Hy, and E7E9-6-ol-11-one-21Hy might be minor sex pheromone components of PAM. Moreover, attractiveness of synthetic pheromone and virgin PAM females needs to be compared to determine whether synthetic pheromone could replace PAM females as trap baits in the program to monitor eradication of exotic PAM in New Zealand. PMID- 15898505 TI - Identification of sex pheromone components of the painted apple moth: a tussock moth with a thermally labile pheromone component. AB - The sex pheromone of the painted apple moth, Teia anartoides (Lymantriidae) was investigated using GC-EAD and GC-MS analysis, derivatization, TLC analysis, and field cage and field trapping bioassays. The major sex pheromone components were identified as (6Z,9Z)-henicosa-6,9-dien-11-one and (6Z,9Z)-henicosa-6,9-diene. Other minor components of pheromone gland extracts included (6Z)-9R, 10S epoxyeicos-6-ene, (6Z)-9R,10S-epoxyhenicos-6-ene, (6Z,9Z)-henicosa-6,9-dien-11 ol, (6Z)-henicos-6-en-11-one, and (6Z, 8E)-henicosa-6,8-dien-11-one, but the roles of these minor components remain equivocal. In field cage and field experiments, a blend of all seven identified components [(6Z,9Z)-henicosa-6,9 dien-11-one (relative amount 100), (6Z,9Z)-henicosa-6,9-diene (100), (6Z)-9R,10S epoxyeicos-6-ene (5), (6Z)-9R,10S-epoxyhenicos-6-ene (10), (6Z,9Z)-henicosa-6,9 dien-11-ol (5), (6Z)-henicos-6-en-11-one (1), and (6Z,8E)-henicosa-6,8-dien-11 one (25)] was as attractive to males as calling females, but tests with blends of the major component(s) with subsets of the minor components did not produce consistent results that unequivocally showed the various minor components to be critical components of the active blend. (6Z,9Z)-henicosa-6,9-dien-11-one is thermally labile and rearranges to (6Z,8E)-henicosa-6,8-dien-1-one and other products at ambient temperature, rendering the synthetic pheromone lure inactive after two days of field exposure. PMID- 15898506 TI - Identification of a sex pheromone component of the geometrid moth Milionia basalis pryeri. AB - A single component in extracts of virgin female Milionia basalis pryeri moths elicited responses from male moth antennae. This compound (ca. 7 ng/female) was identified as (Z,Z)-(3S,4R)-3,4-epoxynonadeca-6,9-diene by GC-MS and NMR analyses, microchemical reactions, and comparative chiral HPLC. In a field test, synthetic (Z,Z)-(3S,4R)-3,4-epoxynonadeca-6,9-diene attracted male moths. The opposite enantiomer, the racemic mixture, and virgin female moths held in small cages attracted no more moths than the solvent controls. PMID- 15898507 TI - The aggregation pheromone of Diorhabda elongata, a biological control agent of saltcedar (Tamarix spp.): identification of two behaviorally active components. AB - The leaf beetle Diorhabda elongata Brulle (Coleoptera: Chrysomelidae) has been introduced as a biological control agent for saltcedars, Tamarix spp., an exotic, invasive weedy tree in the western United State. Gas chromatographic (GC) analysis of volatiles collected from feeding male or female beetles, or saltcedar foliage alone, showed two components produced almost exclusively by males. These compounds elicited responses from antennae of male and female beetles in GC electroantennographic detection (EAD) analyses. The compounds were identified as (2E,4Z)-2,4-heptadienal (1) and (2E,4Z)-2,4-heptadien-1-ol (2) by GC-mass spectrometry (MS), and confirmed with authentic standards. The two compounds were also detected at trace levels from feeding females and foliage controls, but the amounts from feeding males were 8-40 times higher, typically 55-125 ng per day per male. The amounts of 1 and 2 in collections from females did not differ significantly from amounts collected from control foliage. In field trials, 2 as a single component was as attractive as a 1:1 blend of 1 and 2. Compound 1 as a single component was more attractive than controls, but much less attractive than 2 or the blend. Males and females were attracted in about equal numbers, indicating that this is an aggregation pheromone. PMID- 15898508 TI - Responses of the crab Heterozius rotundifrons to heterospecific chemical alarm cues: phylogeny vs. ecological overlap. AB - The big-handed brachyuran crab Heterozius rotundifrons extends the time spent in its anti-predator posture, limb extended posture, if exposed to chemical cues from crushed conspecifics. In this study, we tested whether crabs also respond to chemical cues from crushed heterospecific crabs, and if so, whether phylogenetic relations or ecological overlap is more important in influencing the duration of the anti-predator posture. Chemical cues from two other brachyuran crabs (Cyclograpsus lavauxi and Hemigrapsus sexdentatus), which do not overlap directly in ecological distribution with H. rotundifrons, elicited a duration of the anti predator posture that was indistinguishable from that produced by conspecific chemical cues. In contrast, chemical cues from two anomuran crabs (Petrolisthes elongatus and Pagurus novizealandiae) that overlap in ecological distribution with H. rotundifrons, elicited durations of the antipredator posture that were significantly shorter than those of either conspecifics or more closely related crab species. Thus, phylogenetic relationship seems to be more important than ecological overlap in influencing anti-predator behavior in H. rotundifrons. PMID- 15898509 TI - John B. Simeone 1919-2005. PMID- 15898510 TI - Motorcycle mayhem. PMID- 15898511 TI - Patient satisfaction after laparoscopic antireflux surgery: community achievement. PMID- 15898512 TI - Female patients' sex preferences in selection of gynecologists and surgeons. AB - OBJECTIVES: In this study, the authors investigated sex preferences for gynecologists and surgeons among female patients, and explored potential contributing factors. METHODS: One hundred forty-six female patients were surveyed in a private practice office concerning their sex preferences and past obstetric/gynecologic care. For data comparisons, chi2 or Fisher exact tests were used. RESULTS: Gynecologist sex preferences were similar between male (30%), female (35%), and no sex preferences (35%). Patients who had a female obstetrician at their first delivery or began their gynecologic care with a female were more likely to prefer a female gynecologist. Multiparous patients were more likely to state no preference for a gynecologist. There were no statistical differences in sex preferences when patients were stratified by age, race, educational background, age of first gynecologist visit, or the age at their first delivery. About half of the patients (51%) stated that they preferred a male surgeon; only 3% preferred a female surgeon, and 46% stated they had no preference. CONCLUSIONS: Our investigation demonstrated that women's preferences for a gynecologist were divided equally between preferring a male, a female, and having no preference. Our study did find, however, that about half of the female patients preferred a male surgeon. PMID- 15898513 TI - Racial bias in patient selection of an obstetrician. AB - OBJECTIVE: The objective of this study was to determine racial bias in patient selection of an obstetrician. METHODS: Obstetrical patients referred for genetic counseling at a community hospital were included. Self-reported patient race/ethnicity were compared with obstetrician's race/ethnicity. RESULTS: The patient population (n = 1,519) was 27.8% white, 25% Hispanic, 22.5% black, 20.5% Asian, and 4.3% other. Physician race/ethnicity was 47.8% white, 28.8% Asian, 14.4% Hispanic, and 9% black. Patient race/ethnicity and physician race/ethnicity were correlated (contingency coefficient = 0.54, P < 0.001). White and Asian patients were more likely to select obstetricians of their own racial background (72.7% and 66.6%) than were Hispanic or black patients (36% and 24.6%, P < 0.001). Patients of all races were more likely to be under the care of a physician of their own race than of a different race. CONCLUSIONS: In an urban community with a racially diverse population, there is a strong racial bias in patient selection of an obstetrician. PMID- 15898514 TI - Maternal prepregnant body mass index and weight gain related to low birth weight in South Carolina. AB - OBJECTIVES: The primary purpose of this study was to describe the proportion of low birth weight that could be potentially prevented by programs focusing on maternal prepregnant body mass index (BMI) and/or weight gain during pregnancy. METHODS: In this historic cohort design, study data consisted of birth certificates linked to the Pregnancy Risk Assessment Monitoring System for South Carolina resident women delivering in South Carolina during 1998 and 1999. Statistical analysis was conducted with the use of chi2, population-attributable risk, and logistic regression. The analysis was performed using SUDAAN to accommodate the analysis weight and extrapolate the sample data to the South Carolina state population. RESULTS: Eight percent of the very low birth weight (VLBW) rate in South Carolina can be attributed to inadequate weight gain in pregnancy. Approximately 19% of the state's VLBW rate can be attributed to either underweight or overweight BMI at conception. Women with less than adequate weight gain were 1.4 times more likely to deliver a VLBW baby and 1.9 times more likely to deliver a moderately low birth weight baby as compared with women with adequate weight gain. CONCLUSIONS: Appropriate maternal BMI at conception followed by adequate weight gain during pregnancy may have a substantial influence on reducing the number of low birth weight deliveries. PMID- 15898515 TI - Depression screening in the pregnant soldier wellness program. AB - OBJECTIVES: The purpose of this study was to determine the prevalence of depression among active-duty low-risk pregnant women using the Edinburgh Postnatal Depression Scale. Rates for depression have been reported to be as high as 13% during pregnancy and 12 to 22% postpartum, with postpartum suicidal ideation at nearly 7%. METHODS: From April 2002 through March 2003, 82 women receiving prenatal care at Madigan Army Medical Center completed screening questionnaires during pregnancy and/or postpartum visits. RESULTS: A total of 97 questionnaires were administered (71 antepartum and 26 postpartum). During pregnancy, 24% of individuals screened scored positive. During the postpartum, 19% scored positive. A total of 15 women were screened twice during the study period. One woman was screened twice during pregnancy and the remaining 14 were screened once antepartum and once postpartum. Suicidal ideation was present in 11% of those screened during pregnancy and in 15% postpartum. CONCLUSIONS: Active duty women appear to have a higher rate of depression and suicidal ideation compared with rates in nonmilitary populations; perhaps such screening should become a standard practice. PMID- 15898516 TI - Postpartum hemorrhage after vaginal birth: an analysis of risk factors. AB - OBJECTIVE: To determine, in a single tertiary obstetric hospital, the incidence of and risk factors for postpartum hemorrhage (PPH) after a vaginal birth. METHODS: PPH was defined as measured blood loss greater than 1,000 mL and/or need for a transfusion. RESULTS: Over a 4-year period, 13,868 of 19,476 women delivered vaginally, with a PPH rate of 5.15%. Identified risk factors for PPH were Asian race, maternal blood disorders, prior PPH, history of retained placenta, multiple pregnancy, antepartum hemorrhage, genital tract lacerations, macrosomia (>4 kg), and induction of labor, as well as chorioamnionitis, intrapartum hemorrhage, still birth, compound fetal presentation, epidural anesthesia, prolonged first/second stage of labor, and forceps delivery after a failed vacuum. CONCLUSIONS: Identification of risk factors for PPH after a vaginal delivery may afford prophylactic treatment of such women with reduction of morbidity. PMID- 15898517 TI - Availability and perceived competence of pediatricians to serve as child protection team medical consultants: a survey of practicing pediatricians. AB - OBJECTIVES: The effectiveness of multidisciplinary child protection teams has been demonstrated. This study is an attempt to assess the level of this commitment and the perceived competence of primary care pediatricians to provide this service. METHODS: A questionnaire survey was mailed to primary care pediatricians practicing in the state of Alabama. RESULTS: Among respondents who did not consider themselves competent to conduct sexual abuse or physical abuse examinations, 27% and 19%, respectively, were called on to conduct such examinations. Approximately half of respondents expressed a willingness to serve as consultants under a time commitment obligation of less than 2 hours per week or 1 day per month, and under a reimbursement provision of $200 per examination or less. Respondents recognized a need for, and expressed a desire for, more training in this area. CONCLUSIONS: Primary care pediatricians are willing to serve as multidisciplinary child protection team medical consultants if provided appropriate training and support. PMID- 15898518 TI - Physical and behavioral health of Medicaid children in two southern states. AB - OBJECTIVES: As the primary insurer of children in the United States, Medicaid covers at least one in four US children. Information on the health and behavioral health needs of this group of children is critical to plan, deliver, and monitor services accordingly. METHODS: Parent interview data from a representative sample of Medicaid children in two Southern states were used to generate information from standardized questionnaires on physical health status, chronic illnesses, physical functioning, emotional and behavioral symptoms, and psychosocial functioning. RESULTS: The levels of physical and behavioral health and co occurring problems were higher than other estimates available on the general population. CONCLUSIONS: The high levels of health problems among Medicaid enrolled children need attention in the current struggles over Medicaid reform. Support for improving screening, referral, and integration of services is discussed, as well as the importance of monitoring service system performance in this era of managed care. PMID- 15898519 TI - Trends in Arkansas motorcycle trauma after helmet law repeal. AB - OBJECTIVES: This study aimed to assess the impact of the 1997 Arkansas helmet law repeal on motorcycle registrations, crash and fatality risks, and alcohol involvement in motorcycle crashes. METHODS: Annual motorcycle registration data for the years 1990 through 2001 were obtained from the Arkansas Department of Finance and Administration. These motorcycle registration data were complemented by the motorcycle crash data from the Arkansas State Police Highway Safety Office and motorcycle fatality data for the state of Arkansas from the Fatality Analysis Reporting System. The impact of the repeal on crash rates, helmet usage, and alcohol involvement was assessed through comparisons of data from before (1993 to 1996) and after (1998 to 2001) the repeal. RESULTS: After the repeal, an increase in motorcycle registrations correlated with a marked rise in the total number of crashes and fatalities; however, fatalities per crash remained virtually the same. The proportion of motorcycle fatalities that were not wearing a helmet increased from 47.0% (47/100) before the repeal to 78.2% (104/133) after the repeal (P = 0.001). The overall percentage of fatal motorcycle crashes involving alcohol use remained unchanged after the repeal (37.6% [29/77] to 38.5% [40/104], P = 0.91), but the percentage of fatal crashes involving drinking nonhelmeted drivers increased from 14.2% (11/77) to 33.6% (35/104) (P = 0.003). Inebriated motorcyclists killed in crashes were overwhelmingly non-helmeted (87.5%, 35/40) after the repeal, up from 37.9% (11/29) before the repeal (P < 0.001). CONCLUSIONS: These findings suggest that the repeal of the mandatory helmet law in Arkansas has had a significant adverse effect on road safety. PMID- 15898520 TI - Laparoscopic Nissen fundoplication in a community hospital: patient satisfaction survey. AB - OBJECTIVES: To assess the efficacy of laparoscopic Nissen fundoplication for esophageal reflux in a community hospital. METHODS: One hundred nineteen patients (76%) of 157 patients having laparoscopic fundoplication for symptomatic reflux disease completed a questionnaire. All patients were evaluated with esophagoscopy, esophageal motility, and pH studies. RESULTS: Mean follow-up was 49 months. Ninety-three patients (78%) no longer had heartburn. Seven patients still had daily heart-burn (5.9%). One hundred three patients (86.6%) had no sour regurgitation. Seventy-five patients (63%) had no dysphagia after surgery. Forty two patients had some dysphagia after surgery (36.8%). Six patients (5.0%) had significant dysphagia. Only 4 patients stated that dysphagia interfered with their quality of life. Twenty-six patients (22.8%) thought that belching and bloating were worse, whereas 42 patients (36.8%) thought that belching and bloating were better after surgery. Antacids, histamine-2 antagonist, and proton pump inhibitors were still being used by 12.6%, 14.3%, and 21.8% of patients, respectively. CONCLUSIONS: Eighty-seven patients were completely satisfied (73.1%), 26 patients somewhat satisfied (22.8%), and 6 patients unsatisfied (5.3%) with surgery. Overall satisfaction with the surgery was 94.7%. One hundred five patients (88.2%) would recommend the surgery to others. Laparoscopic Nissen fundoplication can be effectively done in a community hospital setting. PMID- 15898521 TI - Extent and determinants of physician participation in expert witness testimony. AB - OBJECTIVES: In the recent past, law firms have had difficulties in finding physicians to review malpractice cases and serve as expert witnesses. Over the last few years, however, many clinicians have had a decline in their income, causing some physicians to seek alternative sources of revenue such as the review of legal cases. The purpose of this study was to qualify and quantify the extent of physician participation in legal activities. METHODS: We conducted a survey of 1,000 Maryland internal medicine physicians. The survey consisted of 24 items designed to collect information on demographics, practice patterns, income variables, and participation in legal review. RESULTS: Twenty-four percent of our respondents reported engaging in expert witness testimony/review to supplement their incomes. Engagement was significantly associated with internal medicine sub specialty practice, academic practice, a self-perception that personal income was higher than the income of colleagues, and being in practice for 11 to 20 years. Economic factors were not found to be associated with engaging in these activities. CONCLUSIONS: Physician participation in legal review and expert witness activities is significant and appears not to be determined by economic factors. Both the positive effects of such participation on the legal system and the potential ethical concerns require further study. PMID- 15898522 TI - Implications of cyberspace communication: a role for physicians. AB - Through the presentation of three clinical case reports and subsequent discussion, it is demonstrated that physicians must begin to familiarize themselves with the health-related implications of online communication, and must proactively address Internet use as it relates to health and well-being. Included case presentations highlight the following: the established association between those seeking sexual partners through the Internet and an increased risk for sexually transmitted disease; the implications of cyber-communication for young people and concerns related to unsafe online behaviors including sharing identifying information with strangers; the potential use of strategically constructed virtual identities to facilitate sexual exploitation; the impact of accelerated intimacy and disinhibition evident in online communication; and the invasive nature of Internet sexual harassment or bullying. Although it is recognized that most online activities do not negatively affect health, doctors must be prepared to ask patients about Internet use and become involved in educating children, teenagers, and parents about safe online relationships to promote optimal physical, mental, and social health. PMID- 15898523 TI - Systematic review of the risk of uterine rupture with the use of amnioinfusion after previous cesarean delivery. AB - OBJECTIVES: Amnioinfusion is commonly used for the intrapartum treatment of women with pregnancy complicated by thick meconium or oligohydramnios with deep variable fetal heart rate decelerations. Its benefit in women with previous cesarean deliveries is less known. Theoretically, rapid increases in intrauterine volume would lead to a higher risk of uterine rupture. METHODS: Searches of the Cochrane Library from inception to the third quarter of 2001 and MEDLINE, 1966 to November 2001, were performed by using keywords "cesarean" and "amnioinfusion." Search terms were expanded to maximize results. All languages were included. Review articles, editorials, and data previously published in other sites were not analyzed. RESULTS: Four studies were retrieved having unduplicated data describing amnioinfusion in women who were attempting a trial of labor after previous cesarean section. As the studies were of disparate types, meta-analysis was not possible. CONCLUSIONS: The use of amnioinfusion in women with previous cesarean delivery who are undergoing a trial of labor may be a safe procedure, but confirmatory large, controlled prospective studies are needed before definitive recommendations can be made. PMID- 15898524 TI - Lessons learned in the management of thirteen celiac axis injuries. AB - OBJECTIVES: Celiac axis injuries are rare. The purposes of this study were to (1) review institutional experience, (2) determine additive effect on death of associated vessel injuries, and (3) correlate mortality rates with the American Association for the Surgery of Trauma-Organ Injury Scale (AAST-OIS) for abdominal vascular injury. METHODS: This was a retrospective, 132-month study (January 1992 to December 2002) of patients with celiac axis injuries. RESULTS: Thirteen patients were included in the study. Mean revised trauma score was 5.35+/-2.63; mean injury severity score was 25+/-12. The mechanism was penetrating in 12 (92%) and blunt in 1 (8%); 3 of 13 had Emergency Department thoracotomy (100% mortality rate). Treatment included ligation in 11 and primary repair in 1; 1 exsanguinated. Overall survival was 5 of 13 (38%). Adjusted survival excluding patients who had Emergency Department thoracotomy was 5 of 10 patients (50%). Those surviving with isolated injuries included 57% of patients. Mortality rate versus AAST-OIS was grade III, 43% (3 of 7 patients); grade IV, 50% (1 of 2 patients); and grade V, 100% (4 of 4 patients). CONCLUSIONS: Celiac axis injuries are rare. Patients with isolated injuries have better survival rates. Mortality rate correlates well with AAST-OIS for abdominal vascular injury. PMID- 15898525 TI - Hypereosinophilic syndrome: two case reports of contrasting variants. AB - Two cases of hypereosinophilic syndrome that illustrate different variants are reported. Recognition of the specific variant is important because the treatments differ. Patients with hypereosinophilic syndrome should be monitored for evidence of a myeloproliferative process because treatment with tyrosine kinase inhibitors in these patients is effective and may prevent significant morbidity. PMID- 15898526 TI - Giant cell arteritis of the female genital tract: report of a case and review of the literature. AB - A 65-year-old female presented with constitutional symptoms of fever and weight loss with bilateral adnexal masses on physical examination. Total abdominal hysterectomy and bilateral salpingo-oophorectomy were performed and subsequently revealed giant cell arteritis involving numerous small arteries in the ovaries, fallopian tubes, paraovarian and paratubal soft tissues, myometrium, and cervix. After surgery, the patient continued to have constitutional symptoms. Corticosteroid therapy led to a significant improvement and eventual resolution of symptoms. Several similar cases of giant cell arteritis of the female genital tract have been described, both with and without concomitant temporal arteritis. Implications for diagnosis and treatment are discussed. PMID- 15898527 TI - Cricopharyngeal achalasia responsive to balloon dilation in an infant. AB - This report describes an infant with vomiting and significant weight loss attributable to cricopharyngeal achalasia, a rare finding in children. The infant responded to balloon dilation of the upper esophageal sphincter, with resolution of symptoms and return to presymptomatic growth parameters. A brief description of the clinical features, diagnostic evaluation, and treatment options for cricopharyngeal achalasia is included. PMID- 15898528 TI - Wolf in sheep's clothing: advanced Kaposi sarcoma mimicking vulvar abscess. AB - Kaposi sarcoma is a vascular neoplastic disorder that is associated with the acquired immunodeficiency syndrome (AIDS). The causative factor in Kaposi sarcoma is human herpes virus-8. This complication of AIDS has a predilection for homosexual males and is rarely associated with the female AIDS population. However, we present a case of Kaposi sarcoma mimicking the benign and common Bartholin gland abscess. A search of the literature including the MEDLINE database revealed a single report of Kaposi sarcoma presenting as a vulvar mass, but not as a suspected Bartholin abscess. The history, presentations, risk factors, and treatments available for Kaposi sarcoma are also discussed in this report. PMID- 15898529 TI - Worsening of heart failure in Becker muscular dystrophy after nonsteroidal anti inflammatory drugs. AB - A 40-year-old male with Becker muscular dystrophy and cardiac involvement was stable in New York Heart Association class II for 7 years. He then had arthralgia caused by bilateral gonarthrosis. He received diclofenac 100 mg/d and 500 mg/d mephenamine acid. Six weeks later, he was hospitalized because of heart failure (New York Heart Association class IV). Echocardiography revealed biatrial and biventricular dilation, a left ventricular end-diastolic diameter of 82 mm, an ejection fraction of 26%, a severe tricuspid regurgitation, and an elevated pulmonary artery pressure of 60 mm Hg. Nonsteroidal anti-inflammatory drugs were discontinued, and physiotherapy and equipment with a corset were initiated. He improved after treatment with parenteral diuretics, returning to class II. Nonsteroidal anti-inflammatory drugs should be given with caution in Becker muscular dystrophy with cardiac involvement. PMID- 15898530 TI - Severe thrombocytopenia due to hypersplenism successfully treated with partial splenic embolization in preoperative management. AB - Hypersplenism is a known complication of portal hypertension secondary to cirrhosis of the liver. Although thrombocytopenia secondary to hypersplenism does not cause clinically significant hemostatic defect, it may need to be addressed in selective circumstances, such as preoperative preparation for a surgery. This report describes a 30-year-old male with a history of cirrhosis of the liver and hypersplenism who had a recurrence of craniopharyngioma. A platelet count of 40 x 10(9)/L limited his treatment options. A stereotactic injection of radioactive P32 into the tumor was planned but was thought not to be feasible because of the thrombocytopenia. The thrombocytopenia responded favorably to partial splenic embolization, and the patient underwent successful stereotactic injection of radioactive P32 into the tumor. PMID- 15898531 TI - Recurrent arthralgias in a patient with previous Mayaro fever infection. AB - Mayaro fever is an acute, self-limited, febrile, mosquito-borne viral disease manifested by fever, chills, headache, myalgias, and arthralgias. The virus belongs to the family Togaviridae and the genus Alphavirus. Five other mosquito borne viruses have been described as causing a similar dengue-like illness. The virus was first isolated in 1954, and the first epidemics were described in 1955 in Brazil and Bolivia. Other cases have been reported in Suriname, Brazil, Peru, French Guiana, and Trinidad. Up to 10 to 15% of febrile illnesses in endemic areas have been attributed to Mayaro virus. The exact pathogenesis and pathophysiology among humans is unknown. Animal models have demonstrated necrosis of skeletal muscle, periosteum, perichondrial tissues, and evidence of meningitis and encephalitis. All previous cases of Mayaro fever describe a self-limited illness. No reports of recurrent symptoms exist in the literature. This report describes a case of recurrent arthralgias in a military service member presenting to the emergency department. PMID- 15898532 TI - Pedunculated localized fibrous tumor of the pleura presenting as a moving chest mass. AB - Localized fibrous tumor of the pleura is a rare primary pleural neoplasm. The tumors are usually discovered incidentally on routine chest radiography. A change in the lesion's position with respiration and/or posture of the patient indicates a pedunculated pleural lesion. Computed tomography and magnetic resonance imaging reveal the relation of the lesions to adjacent structures. Magnetic resonance imaging is also useful for the characterization of the lesions on the basis of signal features. These tumors usually consist of mature fibrous tissue. PMID- 15898533 TI - Pseudotumor presentation of intracerebral tuberculomas. AB - Diagnosis of tuberculoma is difficult because of its tumorlike aspects. This report describes the case of a male who displayed a hemiplegia revealing an intracranial mass. Neuroimaging was consistent with a glioblastoma; however, the definite diagnosis was a tuberculoma. Clinical features of tuberculomas are nonspecific. Even though the neuroimaging features are sensitive, they are much less specific, with variability related to the tuberculoma course. Investigations leading to the diagnosis are histologic analysis showing a granuloma with or without caseating necrosis, and the microbiologic identification of Mycobacterium tuberculosis. Every intracranial tumor with malignant radiologic and clinical appearance must evoke a suspicion for tuberculoma. PMID- 15898534 TI - Intracerebral bleeding in a patient with neurosarcoidosis while on corticosteroid therapy. AB - Neurosarcoidosis can have variable clinical manifestations. Intracerebral bleeding is very rare in the setting of neurosarcoidosis and has been reported only twice. In this report, we describe a patient with neurosarcoidosis who had intracerebral bleeding while on corticosteroid therapy, despite apparent clinical improvement. PMID- 15898535 TI - Diagnosis of colon and rectal cancer in a large VA Medical Center practice: does fecal occult blood screening make a difference? PMID- 15898536 TI - Unsedated endoscopy. PMID- 15898537 TI - Improving risk factors in diabetic patients participating in a weight management program. PMID- 15898538 TI - The patient's page. PMID- 15898539 TI - Texture and haptic cues in slant discrimination: reliability-based cue weighting without statistically optimal cue combination. AB - A number of models of depth-cue combination suggest that the final depth percept results from a weighted average of independent depth estimates based on the different cues available. The weight of each cue in such an average is thought to depend on the reliability of each cue. In principle, such a depth estimation could be statistically optimal in the sense of producing the minimum-variance unbiased estimator that can be constructed from the available information. Here we test such models by using visual and haptic depth information. Different texture types produce differences in slant-discrimination performance, thus providing a means for testing a reliability-sensitive cue-combination model with texture as one of the cues to slant. Our results show that the weights for the cues were generally sensitive to their reliability but fell short of statistically optimal combination--we find reliability-based reweighting but not statistically optimal cue combination. PMID- 15898540 TI - Metamer sets. AB - If two different surfaces look the same when viewed under a particular light source, then they are called metamers. We show mathematically how one can solve for the whole set of physically realizable natural surface reflectances that relate to the same tristimulus, the metamer set. Our analysis is based on very general linear models of reflectances, coupled with constraints that reflectances should adhere to (e.g., positivity and boundedness). We show that we can recover metamer sets for linear models of an arbitrary high dimension. To illustrate our new algorithm, we provide an example of calculating the metamer set and its manifestation as a mismatch region. Given a single XYZ observed under illuminant D65, we can examine the set of XYZs that would be possible under illuminant A. PMID- 15898541 TI - Statistical properties of color-signal spaces. AB - In applications of principal component analysis (PCA) it has often been observed that the eigenvector with the largest eigenvalue has only nonnegative entries when the vectors of the underlying stochastic process have only nonnegative values. This has been used to show that the coordinate vectors in PCA are all located in a cone. We prove that the nonnegativity of the first eigenvector follows from the Perron-Frobenius (and Krein-Rutman theory). Experiments show also that for stochastic processes with nonnegative signals the mean vector is often very similar to the first eigenvector. This is not true in general, but we first give a heuristical explanation why we can expect such a similarity. We then derive a connection between the dominance of the first eigenvalue and the similarity between the mean and the first eigenvector and show how to check the relative size of the first eigenvalue without actually computing it. In the last part of the paper we discuss the implication of theoretical results for multispectral color processing. PMID- 15898542 TI - Design theories and performance limits of diffractive superresolution elements with the highest sidelobe suppressed. AB - For the application of optical data storage, theories that we present can be used to design a diffractive super-resolution element (DSE) with the highest sidelobe suppressed. A globally optimal solution among general hybrid-type filters can be solved through linear programming. The obtained globally optimal performances set the exact performance limits of a general hybrid-type DSE with the highest sidelobe suppressed. A comparison of our design theories and the previous design methods shows the advantages of the former. PMID- 15898543 TI - Radiometric framework for image mosaicking. AB - Nonuniform exposures often affect imaging systems, e.g., owing to vignetting. Moreover, the sensor's radiometric response may be nonlinear. These characteristics hinder photometric measurements. They are particularly annoying in image mosaicking, in which images are stitched to enhance the field of view. Mosaics suffer from seams stemming from radiometric inconsistencies between raw images. Prior methods feathered the seams but did not address their root cause. We handle these problems in a unified framework. We suggest a method for simultaneously estimating the radiometric response and the camera nonuniformity, based on a frame sequence acquired during camera motion. The estimated functions are then compensated for. This permits image mosaicking, in which no seams are apparent. There is no need to resort to dedicated seam-feathering methods. Fundamental ambiguities associated with this estimation problem are stated. PMID- 15898544 TI - Generation of phase singularity through diffracting a plane or Gaussian beam by a spiral phase plate. AB - We deduce and study an analytical expression for Fresnel diffraction of a plane wave by a spiral phase plate (SPP) that imparts an arbitrary-order phase singularity on the light field. Estimates for the optical vortex radius that depends on the singularity's integer order n (also termed topological charge, or order of the dislocation) have been derived. The near-zero vortex intensity is shown to be proportional to rho2n, where p is the radial coordinate. Also, an analytical expression for Fresnel diffraction of the Gaussian beam by a SPP with nth-order singularity is analyzed. The far-field intensity distribution is derived. The radius of maximal intensity is shown to depend on the singularity number. The behavior of the Gaussian beam intensity after a SPP with second-order singularity (n = 2) is studied in more detail. The parameters of the light beams generated numerically with the Fresnel transform and via analytical formulas are in good agreement. In addition, the light fields with first- and second-order singularities were generated by a 32-level SPP fabricated on the resist by use of the electron-beam lithography technique. PMID- 15898545 TI - Applications of improved first Rayleigh-Sommerfeld method to analyze the performance of cylindrical microlenses with different f-numbers. AB - Based on the previous Letter [Opt. Lett. 29, 2345 (2004)], we significantly extend the applications of the improved first Rayleigh-Sommerfeld method (IRSM1) to analyze the focusing performance of cylindrical micro-lenses for different types of profile (continuous or stepwise), different f-numbers (from f/1.5 to f/0.75), and different polarizations (the TE or TM). A number of performance measures of the cylindrical microlenses, such as the focal spot size, the diffraction efficiency, the real focal position, and the normalized sidelobe power, are studied in detail. We compare numerical results obtained by the IRSM1, by the original first Rayleigh-Sommerfeld method (ORSM1), and by the rigorous boundary element method (BEM). For continuously refractive lenses, the results calculated by the IRSM1 are quite close to those obtained by the BEM; in contrast, the results calculated by the ORSM1 significantly deviate from those obtained from the rigorous BEM. For multilevel diffractive lenses, the IRSM1 also provides much more accurate results than the ORSM1. In addition, compared with the BEM, a notable advantage of the IRSM1 is much lower computer memory and time consumption in computations. PMID- 15898546 TI - Geometric view of adaptive optics control. AB - The objective of an astronomical adaptive optics control system is to minimize the residual wave-front error remaining on the science-object wave fronts after being compensated for atmospheric turbulence and telescope aberrations. Minimizing the mean square wave-front residual maximizes the Strehl ratio and the encircled energy in pointlike images and maximizes the contrast and resolution of extended images. We prove the separation principle of optimal control for application to adaptive optics so as to minimize the mean square wave-front residual. This shows that the residual wave-front error attributable to the control system can be decomposed into three independent terms that can be treated separately in design. The first term depends on the geometry of the wave-front sensor(s), the second term depends on the geometry of the deformable mirror(s), and the third term is a stochastic term that depends on the signal-to-noise ratio. The geometric view comes from understanding that the underlying quantity of interest, the wave-front phase surface, is really an infinite-dimensional vector within a Hilbert space and that this vector space is projected into subspaces we can control and measure by the deformable mirrors and wave-front sensors, respectively. When the control and estimation algorithms are optimal, the residual wave front is in a subspace that is the union of subspaces orthogonal to both of these projections. The method is general in that it applies both to conventional (on-axis, ground-layer conjugate) adaptive optics architectures and to more complicated multi-guide-star- and multiconjugate-layer architectures envisaged for future giant telescopes. We illustrate the approach by using a simple example that has been worked out previously [J. Opt. Soc. Am. A 73, 1171 (1983)] for a single-conjugate, static atmosphere case and follow up with a discussion of how it is extendable to general adaptive optics architectures. PMID- 15898547 TI - Paraxial theory for birefringent lenses. AB - The generalized ray tracing for the extraordinary ray through uniaxial crystals developed by Avendano-Alejo and Stavroudis [J. Opt. Soc. Am. A 19, 1674 (2002)] has been applied to derive paraxial refracting equations. Paraxial equations are derived for three cases where the incident, ordinary, and extraordinary rays lie in the incident plane: (a) the crystal axis is parallel to the optical axis, (b) the crystal axis is orthogonal to the optical axis and lies in the plane of incidence, and (c) the crystal axis is orthogonal to both the optical axis and the incident plane. The paraxial ray-tracing equations for the extraordinary ray are represented by matrix operators. The elements of the matrix system give all the information of the focal points and of the principal points. Gaussian formulas are derived, and some examples are presented. PMID- 15898549 TI - Correlation between intensity and phase in monochromatic light. AB - Analytical expressions to describe the phase gradient of monochromatic light by means of the three-dimensional intensity distribution are derived. With these formulas it is shown that the two-dimensional phase gradient in a plane can be completely determined from noninterferometric intensity measurements if the light propagates strictly in one direction. The analytical expressions are verified by means of numerical investigations on simulated speckle fields, and the results are discussed with respect to common deterministic phase retrieval approaches. PMID- 15898548 TI - Scanning holographic microscopy with transverse resolution exceeding the Rayleigh limit and extended depth of focus. AB - We demonstrate experimentally that the method of scanning holographic microscopy is capable of producing images reconstructed numerically from holograms recorded digitally in the time domain by scanning, with transverse and axial resolutions comparable to those of wide-field or scanning microscopy with the same objective. Furthermore, we show that it is possible to synthesize the point-spread function of scanning holographic microscopy to obtain, with the same objective, holographic reconstructions with a transverse resolution exceeding the Rayleigh limit of the objective up to a factor of 2 in the limit of low numerical aperture. These holographic reconstructions also exhibit an extended depth of focus, the extent of which is adjustable without compromising the transverse resolution. PMID- 15898550 TI - Polarimetric calibration of large-aperture telescopes. II. Subaperture method. AB - A new method for absolute polarimetric calibration of large telescopes is presented. The proposed method is highly accurate and is based on the calibration of a small subaperture, which is then extended to the full system by means of actual observations of an astronomical source. The calibration procedure is described in detail along with numerical simulations that explore its robustness and accuracy. The advantages and disadvantages of this technique with respect to other possible alternatives are discussed. PMID- 15898551 TI - Tunable microwave filter that uses a high-birefringent fiber and a differential group-delay element. AB - We propose and experimentally demonstrate a novel and simple photonic microwave notch filter that uses a high-birefringent fiber that gives a fixed differential group delay (DGD), together with a DGD element that gives a tunable DGD. This configuration overcomes the problems of optical coherence interference and chromatic dispersion, which may occur in schemes that use fiber delay lines or fiber gratings. Also presented is a theoretical analysis for the performance of the microwave filter that uses the present configuration. The present scheme provides a continuous tuning capability for changing the notch frequency. Measured notch rejection is greater than 40 dB. This scheme can operate over a wide wavelength range of the optical carrier. There is good agreement between experiment results and theoretical analysis. PMID- 15898552 TI - Generalizing, optimizing, and inventing numerical algorithms for the fractional Fourier, Fresnel, and linear canonical transforms. AB - By use of matrix-based techniques it is shown how the space-bandwidth product (SBP) of a signal, as indicated by the location of the signal energy in the Wigner distribution function, can be tracked through any quadratic-phase optical system whose operation is described by the linear canonical transform. Then, applying the regular uniform sampling criteria imposed by the SBP and linking the criteria explicitly to a decomposition of the optical matrix of the system, it is shown how numerical algorithms (employing interpolation and decimation), which exhibit both invertibility and additivity, can be implemented. Algorithms appearing in the literature for a variety of transforms (Fresnel, fractional Fourier) are shown to be special cases of our general approach. The method is shown to allow the existing algorithms to be optimized and is also shown to permit the invention of many new algorithms. PMID- 15898553 TI - Fast numerical algorithm for the linear canonical transform. AB - The linear canonical transform (LCT) describes the effect of any quadratic phase system (QPS) on an input optical wave field. Special cases of the LCT include the fractional Fourier transform (FRT), the Fourier transform (FT), and the Fresnel transform (FST) describing free-space propagation. Currently there are numerous efficient algorithms used (for purposes of numerical simulation in the area of optical signal processing) to calculate the discrete FT, FRT, and FST. All of these algorithms are based on the use of the fast Fourier transform (FFT). In this paper we develop theory for the discrete linear canonical transform (DLCT), which is to the LCT what the discrete Fourier transform (DFT) is to the FT. We then derive the fast linear canonical transform (FLCT), an N log N algorithm for its numerical implementation by an approach similar to that used in deriving the FFT from the DFT. Our algorithm is significantly different from the FFT, is based purely on the properties of the LCT, and can be used for FFT, FRT, and FST calculations and, in the most general case, for the rapid calculation of the effect of any QPS. PMID- 15898554 TI - Iterative zonal wave-front estimation algorithm for optical testing with general shaped pupils. AB - An iterative zonal wave-front estimation algorithm for slope or gradient-type data in optical testing acquired with regular or irregular pupil shapes is presented. In the mathematical model proposed, the optical surface, or wave-front shape estimation, which may have any pupil shape or size, shares a predefined wave-front estimation matrix that we establish. Owing to the finite pupil of the instrument, the challenge of wave front shape estimation in optical testing lies in large part in how to properly handle boundary conditions. The solution we propose is an efficient iterative process based on Gerchberg-type iterations. The proposed method is validated with data collected from a 15 x 15-grid Shack Hartmann sensor built at the Nanjing Astronomical Instruments Research Center in China. Results show that the rms deviation error of the estimated wave front from the original wave front is less than lambda/130-lambda/150 after approximately 12 iterations and less than lambda/100 (both for lambda = 632.8 nm) after as few as four iterations. Also, a theoretical analysis of algorithm complexity and error propagation is presented. PMID- 15898555 TI - Discontinuous spectral element method modeling of optical coupling by whispering gallery modes between microcylinders. AB - We introduce a high-order time-domain discontinuous spectral element method for the study of the optical coupling by evanescent whispering gallery modes between two microcylinders, the building blocks of coupled resonator optical waveguide devices. By using the discontinuous spectral element method with a Dubiner orthogonal polynomial basis on triangles and a Legendre nodal orthogonal basis on quadrilaterals, we conduct a systematic study of the optical coupling by whispering gallery modes between two microcylinders and demonstrate the successful coupling between the microcylinders and also the dependence of such a coupling on the separation and the size variation of the microcylinders. PMID- 15898556 TI - Optimal design of achromatic true zero-order waveplates using twisted nematic liquid crystal. AB - Phase retarders usually show strong wavelength dependence. A novel and simple configuration with the combination of two twisted nematic liquid-crystal cells is proposed for the design of a true zero-order achromatic quarter-wave plate. The present optimization method considers the material dispersion. Simulation computations show a good achromatic behavior of the optimized waveplate. Compared with other types of broadband quarter-wave plates, the present device is compatible with classical liquid-crystal displays and can be expected to be used in precision polarimeters with low cost and enhanced light efficiency. PMID- 15898557 TI - Reflective liquid-crystal displays with asymmetric incident and exit angles. AB - The 2 x 2 matrix methods are extended to calculate the optical behaviors of reflective liquid-crystal displays with asymmetric incident and exit angles. Both the unfolding method and the backward-eigenwave method are employed to derive the 2 x 2 matrix representations. The simulation results for symmetric incident and exit angles from these two methods are identical and agree well with those obtained from the 4 x 4 matrix method when the air-panel surface reflections are neglected. Further, the derived 2 x 2 matrix methods are applied to the asymmetric cases with different incident and exit angles. The simulated results on the normally black vertical alignment and normally white mixed-mode twisted nematic reflective displays show reasonably good agreement with the reported experimental data. In addition, a rubbing effect related to contrast values is observed and analyzed in asymmetric reflective cases. We also find that this effect has a significant influence on the contrast ratios once the difference between the incident and exit angles becomes large. PMID- 15898558 TI - Electromagnetic field for a focused light sheet incident on a plane surface. AB - A solution procedure is developed for the determination of the electromagnetic field that results from the interaction of a focused light sheet with a plane surface. The effects of angle of incidence, relative index of refraction, polarization, and incident light sheet profile on the resulting electromagnetic field distribution are demonstrated. PMID- 15898559 TI - Simple interferometric technique for generation of a radially polarized light beam. AB - We present a theoretical and experimental investigation of an interferometric technique for converting a linearly polarized Gaussian beam into a radially polarized doughnut beam. The experimental setup accomplishes the coherent summation of two orthogonally polarized TEM01 and TEM10 beams that are obtained from the transformation of a TEM00 beam by use of a simple binary diffractive optical element. We have shown that the degree of radial polarization is maximum at a given distance from the interferometer output port that depends on the diameter of the incident beam at the interferometer input port. PMID- 15898560 TI - Generating Bessel beams by use of localized modes. AB - We propose a novel method for generating both propagating and evanescent Bessel beams. To generate propagating Bessel beams we propose using a pair of distributed Bragg reflectors (DBRs) with a resonant point source on one side of the system. Those modes that couple with the localized modes supported by the DBR system will be selectively transmitted. This is used to produce a single narrow band of transmission in kappa space that, combined with the circular symmetry of the system, yields a propagating Bessel beam. We present numerical simulations showing that a propagating Bessel beam with central spot size of approximately 0.5lambda0 can be maintained for a distance in excess of 3000lambda0. To generate evanescent Bessel beams we propose using transmission of a resonant point source through a thin film. A transmission resonance is produced as a result of the multiple scattering occurring between the interfaces. This narrow resonance combined with the circular symmetry of the system corresponds to an evanescent Bessel beam. Because propagating modes are also transmitted, although the evanescent transmission resonance is many orders of magnitude greater than the transmission for the propagating modes, within a certain distance the propagating modes swamp the exponentially decaying evanescent ones. Thus there is only a certain regime in which evanescent Bessel beams dominate. However, within this regime the central spot size of the beam can be made significantly smaller than the wavelength of light used. Thus evanescent Bessel beams may have technical application, in high-density recording for example. We present numerical simulations showing that with a simple glass thin film an evanescent Bessel beam with central spot size of approximately 0.34lambda0 can be maintained for a distance of 0.14lambda0. By choice of different material parameters, the central spot size can be made smaller still. PMID- 15898561 TI - Huygens description of resonance phenomena in subwavelength hole arrays. AB - We develop a point-scattering approach to the plane-wave optical transmission of subwavelength metal hole arrays. We present a real-space description instead of the more conventional reciprocal-space description; this naturally produces interfering resonant features in the transmission spectra and makes explicit the tensorial properties of the transmission matrix. We give transmission spectra simulations for both square and hexagonal arrays; these can be evaluated at arbitrary angles and polarizations. PMID- 15898562 TI - Quantum cryptography with perfect multiphoton entanglement. AB - Multiphoton entanglement in the same polarization has been shown theoretically to be obtainable by type-I spontaneous parametric downconversion (SPDC), which can generate bright pulses more easily than type-II SPDC. A new quantum cryptographic protocol utilizing polarization pairs with the detected type-I entangled multiphotons is proposed as quantum key distribution. We calculate the information capacity versus photon number corresponding to polarization after considering the transmission loss inside the optical fiber, the detector efficiency, and intercept-resend attacks at the level of channel error. The result compares favorably with all other schemes employing entanglement. PMID- 15898563 TI - A view from the cliff. PMID- 15898564 TI - It's a grey matter. PMID- 15898565 TI - Anterior repositioning appliance therapy for TMJ disorders: specific symptoms relieved and relationship to disk status on MRI. AB - Forty-eight (48) consecutive patients seeking treatment in a referral based practice for complex chronic painful temporomandibular joint (TMJ) disease were enrolled in a prospective study to assess specific symptom relief from anterior repositioning appliance (ARA) therapy and the relationship between specific symptom relief and the status of the TMJ disk. Each patient was assessed on 86 symptoms based upon whether each symptom was present before treatment and absent, better, unchanged or worse after Maximum Medical Improvement (MMI). The most common symptom was occipital cephalalgia (94%). The least common symptom was pain and burning of tongue (8%). A profile of a temporomandibular disorder (TMD) patient was developed. The typical TMD patient has cephalalgia, mainly in the occipital, temporal and frontal region, pain upon chewing food, pain upon opening and closing the mouth, TMJ pain, pain in the back of the neck and difficulty chewing food. Before treatment, patients with bilateral displaced disks had more symptoms than those with unilateral displaced disks and the opposite side normal. After MMI, the maximum benefit (percent of pretreatment symptoms relieved) was found in patients with normal or recaptured disks. The minimum occurred in patients whose disks did not recapture with therapy. ARA therapy improved or eliminated symptoms in all patients in the study. PMID- 15898566 TI - Introduction to through-transmission alveolar ultrasonography (TAU) in dental medicine. AB - Through-transmission alveolar ultrasonography (TAU) is a novel imaging modality in dental medicine. A brief introduction to through-transmission ultrasonography (TTU) is followed by a description of the first commercially available TAU device, the Cavitat CAV 4000 (Cavitat Medical Technologies, Inc., Alba, TX). Recent associations between systemic osteoporosis, oral osteoporosis, periodontal diseases, and cardiovascular diseases underline the importance of early detection and treatment of oral cancellous bone pathologies associated with low bone density (LBD), such as regional ischemic osteoporosis, chronic nonsuppurative osteomyelitis, bone marrow edema, and cavitational ischemic osteonecrosis (osteocavitation). While the impact of osteoporosis on maxillofacial bones is acknowledged, there is a lack of reliable prevalence rate, and the National Institutes of Health (NIH) recommend that more attention should be paid to skeletal health, especially in persons with conditions known to be associated with secondary osteoporosis. TAU, a safe and effective imaging modality, can be a valuable tool in research as well as for the clinical assessment of alveolar cancellous bone pathologies associated with LBD and ischemia. PMID- 15898567 TI - A change of occlusal conditions after splint therapy for bruxers with and without pain in the masticatory muscles. AB - Bruxism has been suggested as an initiating or perpetuating factor in a certain subgroup of temporomandibular disorders (TMD), however, the exact association between bruxism and TMD remains unclear. This study aimed to demonstrate the difference in responses between bruxism and a subgroup of TMD to a full-arch maxillary stabilization splint from the standpoint of an occlusal condition. This study was conducted to verify the null hypothesis that there were no differences between bruxer groups with and without myofascial pain (MFP) with respect to the changes in occlusal conditions after the use of a splint. Thirty bruxers with MFP and 30 without MFP participated. Occlusal conditions were examined before and after splint therapy, and occlusal changes following the use of a splint were compared between the two groups. The frequency of occlusal changes after splint therapy was significantly higher in the MFP bruxer group than the non-MFP bruxer group (p < 0.05) for the occlusal conditions investigated in the present study. However, no statistical differences were found with regard to each occlusal condition. This result may show the variety of splint effects and may demonstrate a heterogeneous aspect to bruxism and myofascial pain. PMID- 15898568 TI - The influence of an experimentally-induced malocclusion on vertebral alignment in rats: a controlled pilot study. AB - There is a growing interest in the relationship between occlusion and posture because of a greater incidence of neck and trunk pain in patients with occlusal dysfunction. The study was designed to verify whether an alteration of the spinal column alignment may be experimentally induced in rats as a consequence of altering dental occlusion and also to investigate whether the spinal column underwent any further changes when normal occlusion was then restored. Thirty rats were divided into two groups. Fifteen (15) rats (test group) wore an occlusal bite pad made of composite resin on the maxillary right first molar for a week (T1). The same rats wore a second composite bite pad for another week on the left first molar in order to rebalance dental occlusion (T2). Fifteen rats were included in an untreated control group. All the rats underwent total body radiographs at T0 (before the occlusal pad was placed), at T1 (one week after application of a resin occlusal bite pad on the maxillary left first molar) and at T2 (one week after application of a second resin occlusal bite pad on the maxillary right first molar). A scoliotic curve developed in all the test rats at T1. There were no alterations of spinal position observed in any of the control rats. Additionally, the spinal column returned to normal condition in 83% of the test rats when the balance in occlusal function was restored. The alignment of the spinal column seemed to be influenced by the dental occlusion. PMID- 15898569 TI - Reproducibility of surface EMG in the human masseter and anterior temporalis muscle areas. AB - The aim of this study was to test the hypothesis that surface electromyography (sEMG) recordings, made at mandibular rest position from the masseter and temporalis anterior areas, are intra- and inter-session reproducible. A template was designed and built to permit the correct electrode placement from one session to the next session. A sample of 18 subjects was examined. Two groups, homogeneous for age, sex, and craniofacial morphology were selected. The first group included asymptomatic subjects with no signs or symptoms of temporomandibular joint dysfunction (TMD) and the second group included patients suffering from muscle-related TMD. Data were obtained from different sEMG recordings made at mandibular rest position in the same session and in different sessions, repositioning the electrodes using a template designed for that purpose. The electromyograph used in this, study is part of the EMG K6-I Win Diagnostic System. Results showed that reproducibility of sEMG signals from the masseter and anterior temporalis areas at mandibular rest position is possible. PMID- 15898570 TI - Incidence of systemic joint hypermobility and temporomandibular joint hypermobility in pregnancy. AB - The purpose of this study was to establish a possible correlation between systemic hypermobility and temporomandibular hypermobility during pregnancy. One hundred (100) healthy pregnant women were evaluated: 7% in the first trimester (1T), 38% in the second trimester (2T), and 55% in the third trimester (3T) of gestation. In the series, the authors analyzed systemic joint hypermobility (SJH), range of mandibular movement (MMR), head and shoulder posture, head lateralization, and the presence of noise, pain, and parafunction in the temporomandibular joint. They observed that pain is present to a mild degree mostly in the head and ears of all pregnant women who presented with pain. Most of the subjects had some type of parafunction, but only 42.8% had noises. Mild SJH was seen in 50% of the 2T and 3T subjects, and in 28.5% of 1T subjects. Mild mandibular hypermobility was found for jaw opening (46%) and lateralization to the right (44%) or to the left (46%). Most of the subjects had hypomobility for jaw protrusion and retraction. The subjects had head protrusion and anterior posture as a result of the change in their center of gravity brought about by pregnancy. The authors found no association between systemic joint hypermobility (SJH) and temporomandibular hypermobility, although hormonal changes and complex factors during pregnancy may represent a risk factor for both types of mobility change. PMID- 15898571 TI - Double contours and craniofacial morphology. AB - This study was done to try to clarify the relationship between double contours and craniofacial morphology. The study sample included 56 pre-orthodontic patients with signs and symptoms of temporomandibular joint (TMJ) disorders. A comparison of craniofacial structures was done on 32 subjects with bilateral condylar bone change (BBC group: 28 female and 4 male) and 24 subjects with no condylar bone change (NBC group: 21 female and 3 male). The BBC showed significantly greater antegonial and ramus notch depths, as well as significantly more retruded mandibles, shorter ramus heights, and larger mandibular plane angles than the NBC. BBC subjects with bilateral double contours showed significantly more retruded mandibles, larger antegonial notch depth, and shorter lengths from the Sella to the Articulare than BBC subjects with no double contours. The study showed that important relationships exist between the presence of double contours and mandibular morphology and TMJ position. PMID- 15898572 TI - Long-term skeletal and dental changes in patients with cleft lip and palate after maxillary distraction: a report of three cases treated with a rigid external distraction device. AB - We examined long-term skeletal and dental changes in three patients with cleft lip and palate who underwent maxillary distraction using a rigid external distraction device. Two were children, and one was an adult. Changes in the overjet (OJ), overbite (OB), and positions of the anterior nasal spine (ANS), upper incisors (U1), pogonion (Pog), and lower incisors (L1) were measured on preoperative to 36 months postoperative lateral-cephalograms. In the adult, the positions of all examination points were relatively stable from 6 to 36 months postoperatively, and the OJ and OB were maintained at over 2 mm at 36 months. In the children, the positions of ANS and U1 changed inferiorly, Pog and L1 changed anteroinferiorly, and OJ and OB tended to decrease from 6 to 36 months postoperatively. Long-term skeletal and dental stability following maxillary distraction was relatively well maintained in the adult patient. In the children, the maxillomandibular growth was observed after maxillary distraction, but the mandibular overgrowth might have been inhibited by the correction of class III dentoskeletal deformity resulting from the maxillary distraction. PMID- 15898573 TI - Evolution. PMID- 15898574 TI - Imaging center emergency: who has the back-ups? AB - On first glance, the issue of data back-up appears to be straightforward and relatively uncomplicated since procedures will already be in place. However, as the related risks and requirements are included in the equation, the process of meeting HIPAA requirements in this area becomes more complicated. Data back-up is an important part of disaster and emergency mode operations planning adding to the challenge of meeting the Security Rule's April 21, 2005, compliance date. PMID- 15898575 TI - Boss management. PMID- 15898576 TI - Image display in the operating room: eliminating the barriers in the transition to filmless radiology. AB - As hospitals endeavor to transition from film-based radioloogy to electronic or filmless radiology, one limitation is an effective means for accessing the electronic image archive during surgical procedures. The dependency on using reference images during surgical procedures is a critical function. Scott & White Memorial Hospital in Temple, TX, has been progressively moving toward an electronic paradigm for access to medical information. As the radiology department began to eliminate film as a medium for image presentation and image archiving, it was realized that the hospital needed to provide an electronic solution for the display of images in the operating room (OR) as reference during the surgical procedure. The goals in this project were, therefore, multifold: provide electronic access to images and image files directly within the operating suites, eliminate lost films, and reduced delays caused by lost or unavailable films. The end solution utilized the same Web-based software for all devices, but varied the hardware to meet the individual's or group's needs. The success in this project was not contained to cost savings in radiology, which was realized by reducing film library personnel and eliminating films printed specifically for the surgical environment, but also in greater magnitude for the hospital in improving efficiency of the OR support staff and by directly stimulating a reduction in the average OR time needed for the surgical procedures. PMID- 15898577 TI - Improving medical imaging report turnaround times: the role of technolgy. AB - At Southern Ohio Medical Center (SOMC), the medical imaging department and the radiologists expressed a strong desire to improve workflow. The improved workflow was a major motivating factor toward implementing a new RIS and speech recognition technology. The need to monitor workflow in a real-time fashion and to evaluate productivity and resources necessitated that a new solution be found. A decision was made to roll out both the new RIS product and speech recognition to maximize the resources to interface and implement the new solution. Prior to implementation of the new RIS, the medical imaging department operated in a conventional electronic-order-entry to paper request manner. The paper request followed the study through exam completion to the radiologist. SOMC entered into a contract with its PACS vendor to participate in beta testing and clinical trials for a new RIS product for the US market. Backup plans were created in the event the product failed to function as planned--either during the beta testing period or during clinical trails. The last piece of the technology puzzle to improve report turnaround time was voice recognition technology. Speech recognition enhanced the RIS technology as soon as it was implemented. The results show that the project has been a success. The new RIS, combined with speech recognition and the PACS, makes for a very effective solution to patient, exam, and results management in the medical imaging department. PMID- 15898578 TI - Images anytime, anywhere: they'll love you for it ... or not. PMID- 15898579 TI - The race for the Mercedes. PMID- 15898580 TI - Reinventing radiology reimbursement. AB - Lee Memorial Health System (LMHS), located in southwest Florida, consists of 5 hospitals, a home health agency, a skilled nursing facility, multiple outpatient centers, walk-in medical centers, and primary care physician offices. LMHS annually performs more than 300,000 imaging procedures with gross imaging revenues exceeding dollar 350 million. In fall 2002, LMHS received the results of an independent audit of its IR coding. The overall IR coding error rate was determined to be 84.5%. The projected net financial impact of these errors was an annual reimbursement loss of dollar 182,000. To address the issues of coding errors and reimbursement loss, LMHS implemented its clinical reimbursementspecialist (CRS) system in October 2003, as an extension of financial services' reimbursement division. LMHS began with CRSs in 3 service lines: emergency department, cardiac catheterization, and radiology. These 3 CRSs coordinate all facets of their respective areas' chargemaster, patient charges, coding, and reimbursement functions while serving as a resident coding expert within their clinical areas. The radiology reimbursement specialist (RRS) combines an experienced radiologic technologist, interventional technologist, medical records coder, financial auditor, reimbursement specialist, and biller into a single position. The RRS's radiology experience and technologist knowledge are key assets to resolving coding conflicts and handling complex interventional coding. In addition, performing a daily charge audit and an active code review are essential if an organization is to eliminate coding errors. One of the inherent effects of eliminating coding errors is the capturing of additional RVUs and units of service. During its first year, based on account level detail, the RRS system increased radiology productivity through the additional capture of just more than 3,000 RVUs and 1,000 additional units of service. In addition, the physicians appreciate having someone who "keeps up with all the coding changes" and looks out for the charges. By assisting a few physicians' staff with coding questions, providing coding updates, and allowing them to sit in on educational sessions, at least 2 physicians have transferred some their volume to LMHS from a competitor. The provision of a "clean account," without coding errors, allows the biller to avoid the rework and billing delays caused by coding issues. During the first quarter of the RRS system, the billers referred an average of 9 accounts per day for coding resolution. During the fourth quarter of the system, these referrals were reduced to less than one per day. Prior to the RRS system, resolving these issues took an average of 4 business days. Now the conflicts are resolved within 24 hours. PMID- 15898581 TI - Leading edge, bleeding edge. PMID- 15898582 TI - The top 10 reasons I'm glad I became an X-ray tech. PMID- 15898583 TI - ER radiology: keeping the front door open. PMID- 15898584 TI - The new breed. PMID- 15898585 TI - Chelsea Clinton shows independence at Monitor. PMID- 15898586 TI - Immigrant workforce. Needed by the NHS but denigrated by the politicians. PMID- 15898587 TI - On suspicious minds. PMID- 15898588 TI - Data briefing. NHS spending against GDP. PMID- 15898589 TI - Clinical management. Nervous energy. AB - Neurological conditions account for a fifth of all acute hospital admissions. The long-term neurological conditions NSF takes a structured approach to improving the patient journey. The healthcare commission will have a crucial role in ensuring implementation. PMID- 15898590 TI - Good management. Dodge a mid-life illness. PMID- 15898591 TI - Disability access. A level playing field. PMID- 15898592 TI - Transformation. Ch-ch-changes. PMID- 15898593 TI - The great state health-care giveaway. PMID- 15898594 TI - Socialized medicine? From republicans? PMID- 15898595 TI - The quest for custom cures. PMID- 15898596 TI - Soul of the new gene machines. PMID- 15898597 TI - [Seen from the bottom--colonoscopy in Denmark]. PMID- 15898598 TI - [Probiotics for treating infectious diarrhea. A survey of a Cochrane review]. PMID- 15898599 TI - [Intravitreal triamcinolone therapy. New therapeutic alternative for patients with vision-threatening eye diseases]. PMID- 15898600 TI - [Fulvestrant]. PMID- 15898601 TI - [The quality of colonoscopy in Denmark. Results from a European quality study]. PMID- 15898602 TI - [Quality improvement of colonoscopy in Denmark]. PMID- 15898603 TI - [Appropriateness of colonoscopy. Are European standards applicable to a Danish population?]. PMID- 15898604 TI - [Complications after stoma closure. A retrospective study of 11 years' experience]. PMID- 15898605 TI - [Ten years' results of endoscopic treatment of Zenker's diverticulum with laser evaporation--evaluation of patient satisfaction]. PMID- 15898606 TI - [Does the administration of dipyridamole affect the ventilatory capacity of patients with chronic obstructive lung disease?]. PMID- 15898607 TI - [Cohort study of sibling effect and infectious diseases on the development of atopic dermatitis--secondary publication]. PMID- 15898608 TI - [Ectopic gastric mucosa in the rectum]. PMID- 15898609 TI - [Congenital nephrogenic diabetes insipidus. A four-month-old girl with delayed psychomotor development]. PMID- 15898610 TI - [Noninvasive fetal RHD genotyping in a previously Rh-isoimmunized pregnant woman]. PMID- 15898611 TI - [Mesalazine is effective in Crohn disease]. PMID- 15898612 TI - [Mammography screening--one more time]. PMID- 15898613 TI - [Is there any documentation for Lyrica in the treatment of painful diabetic neuropathy?]. PMID- 15898614 TI - [Green amniotic fluid]. PMID- 15898615 TI - [Effect of mammography screening for breast cancer]. PMID- 15898616 TI - [Definition of unconventional medicine--is it possible?]. PMID- 15898617 TI - An artificial neural network approach for predicting architectural speech security. PMID- 15898619 TI - Design and optimization of a noise reduction system for infrasonic measurements using elements with low acoustic impedance. AB - The implementation of the infrasound network of the International Monitoring System (IMS) for the enforcement of the Comprehensive Nuclear-Test-Ban Treaty (CTBT) increases the effort in the design of suitable noise reducer systems. In this paper we present a new design consisting of low impedance elements. The dimensioning and the optimization of this discrete mechanical system are based on numerical simulations, including a complete electroacoustical modeling and a realistic wind-noise model. The frequency response and the noise reduction obtained for a given wind speed are compared to statistical noise measurements in the [0.02-4] Hz frequency band. The effects of the constructive parameters-the length of the pipes, inner diameters, summing volume, and number of air inlets are investigated through a parametric study. The studied system consists of 32 air inlets distributed along an overall diameter of 16 m. Its frequency response is flat up to 4 Hz. For a 2 m/s wind speed, the maximal noise reduction obtained is 15 dB between 0.5 and 4 Hz. At lower frequencies, the noise reduction is improved by the use of a system of larger diameter. The main drawback is the high frequency limitation introduced by acoustical resonances inside the pipes. PMID- 15898618 TI - Estimates of auditory filter phase response at and below characteristic frequency. PMID- 15898620 TI - Analysis of the sound field in finite length infinite baffled cylindrical ducts with vibrating walls of finite impedance. AB - This paper describes an analytical model of finite cylindrical ducts with infinite flanges. This model is used to investigate the sound radiation characteristics of the gradient coil system of a magnetic resonance imaging (MRI) scanner. The sound field in the duct satisfies both the boundary conditions at the wall and at the open ends. The vibrating cylindrical wall of the duct is assumed to be the only sound source. Different acoustic conditions for the wall (rigid and absorptive) are used in the simulations. The wave reflection phenomenon at the open ends of the finite duct is described by general radiation impedance. The analytical model is validated by the comparison with its counterpart in a commercial code based on the boundary element method (BEM). The analytical model shows significant advantages over the BEM model with better numerical efficiency and a direct relation between the design parameters and the sound field inside the duct. PMID- 15898621 TI - Acoustical measurement of the shear modulus for thin porous layers. AB - Simulations performed with the Biot theory show that for thin porous layers, a shear mode of the structure can be induced by a point-source in air located close to the layer. The simulations show that this mode is present around frequencies where the quarter wavelength of the shear Biot wave is equal to the thickness of the samples and show that it can be acoustically detected from the fast variations with frequency of the location of a pole of the reflection coefficient close to grazing incidence. The mode has been detected with this method for two reticulated plastic foams. For one of the foams studied, the velocity and the damping of the Rayleigh wave have been measured on a thicker layer of the same medium at higher frequencies, giving a real part of the shear modulus close to the one obtained from the measured location of the pole. The strong coupling of the shear mode with the acoustic field in air allows the measurement of the shear modulus without mechanical excitation. PMID- 15898622 TI - Computation of scattering from clusters of spheres using the fast multipole method. AB - A T-matrix based method of solution of the multiple scattering problem was presented by the authors [J. Acoust Soc. Am. 112, 2688-2701 (2002)]. This method can be applied to the computation of relatively small problems, since the number of operations required grows with the number of spheres N as O(N3), and with the sixth power of the wave number. The use of iterative techniques accelerated using the fast multipole method (FMM) can accelerate this solution, as presented by Koc and Chew [J. Acoust. Soc. Am. 103, 721-734 (1998)] originally. In this study we present a method that combines preconditioned Krylov subspace iterative techniques, FMM accelerated matrix vector products, a novel FMM-based preconditioner, and fast translation techniques that enable us to achieve an overall algorithm in which the cost of the matrix-vector multiplication grows with N as O (N logN) and with the third power of the wave number. We discuss the convergence of the iterative techniques, selection of the truncation number, errors in the solution, and other issues. The results of the solution of test problems obtained with the method for N approximately 10(2)-10(4) for different wave numbers are presented. PMID- 15898623 TI - Modeling of phased array transducers. AB - Phased array transducers are multi-element transducers, where different elements are activated with different time delays. The advantage of these transducers is that no mechanical movement of the transducer is needed to scan an object. Focusing and beam steering is obtained simply by adjusting the time delay. In this paper the DPSM (distributed point source method) is used to model the ultrasonic field generated by a phased array transducer and to study the interaction effect when two phased array transducers are placed in a homogeneous fluid. Earlier investigations modeled the acoustic field for conventional transducers where all transducer points are excited simultaneously. In this research, combining the concepts of delayed firing and the DPSM, the phased array transducers are modeled semi-analytically. In addition to the single transducer modeling the ultrasonic fields from two phased array transducers placed face to face in a fluid medium is also modeled to study the interaction effect. The importance of considering the interaction effect in multiple transducer modeling is discussed, pointing out that neighboring transducers not only act as ultrasonic wave generators but also as scatterers. PMID- 15898624 TI - Lamb mode conversion at edges. A hybrid boundary element-finite-element solution. AB - Two general and flexible numerical techniques based on the finite-element and boundary element methods developed by the authors in a previous paper are applied to study Lamb wave propagation in multilayered plates and Lamb mode conversion at free edges for frequencies beyond the first cutoff frequency. Both techniques are supported by a meshing criterion which guarantees the accuracy of the results when a condition is fulfilled. A finite-element formulation is directly applicable to study Lamb wave propagation and reflection by simple obstacles such as a flat edge. In order to tackle Lamb wave diffraction problems by defects with more complex geometries, a hybrid boundary element-finite-element formulation is used. This technique provides a major improvement with respect to the only previous boundary element application on Lamb waves: the connecting boundary might be placed as close to the reflector as desired, reducing greatly the requirement on mesh size. Two main application problems on practical metallic plates are studied and compared with reported numerical, theoretical, and experimental results: (1) Lamb wave propagation in degraded titanium diffusion bonds, and (2) Lamb mode conversion at inclined or perpendicular free edges of steel plates for frequencies beyond the first cutoff frequency. PMID- 15898625 TI - Dispersion of waves in porous cylinders with patchy saturation: formulation and torsional waves. AB - Laboratory experiments on wave propagation through saturated and partially saturated porous media have often been conducted on porous cylinders that were initially fully saturated and then allowed to dry while continuing to acquire data on the wave behavior. Since it is known that drying typically progresses from outside to inside, a sensible physical model of this process is concentric cylinders having different saturation levels-the simplest example being a fully dry outer cylindrical shell together with a fully wet inner cylinder. We use this model to formulate the equations for wave dispersion in porous cylinders for patchy saturation (i.e., drainage) conditions. In addition to multiple modes of propagation obtained numerically from these dispersion relations, we find two distinct analytical expressions for torsional wave modes. We solve the resulting torsional wave dispersion relation for two examples: Massillon sandstone and Sierra White granite. One essential fact that comes to light during the analysis is that the effective shear moduli of the gas- and liquid-saturated regions must differ, otherwise it is impossible to account for the laboratory torsional wave data. Furthermore, the drainage analysis appears to give improved qualitative and quantitative agreement with the data for both of the materials considered. PMID- 15898626 TI - A first-order statistical smoothing approximation for the coherent wave field in random porous random media. AB - An important dissipation mechanism for waves in randomly inhomogeneous poroelastic media is the effect of wave-induced fluid flow. In the framework of Biot's theory of poroelasticity, this mechanism can be understood as scattering from fast into slow compressional waves. To describe this conversion scattering effect in poroelastic random media, the dynamic characteristics of the coherent wavefield using the theory of statistical wave propagation are analyzed. In particular, the method of statistical smoothing is applied to Biot's equations of poroelasticity. Within the accuracy of the first-order statistical smoothing an effective wave number of the coherent field, which accounts for the effect of wave-induced flow, is derived. This wave number is complex and involves an integral over the correlation function of the medium's fluctuations. It is shown that the known one-dimensional (1-D) result can be obtained as a special case of the present 3-D theory. The expression for the effective wave number allows to derive a model for elastic attenuation and dispersion due to wave-induced fluid flow. These wavefield attributes are analyzed in a companion paper. PMID- 15898627 TI - Linear and nonlinear propagation of higher order modes in hard-walled circular ducts containing a real gas. AB - This paper deals with the linear and nonlinear propagation of sound waves through a real gas contained in a circular tube with rigid, isothermal walls. Special emphasis is placed on the asymptotically correct treatment of the higher order modes and their interaction with the acoustic boundary layer. In the first part, a linear perturbation analysis is carried out to calculate the correction terms arising from the viscothermal damping mechanisms present in the system. In extension to previous work, the propagation length is assumed to be so large that the exponentially growing boundary layer effects do not only affect the second order terms of the sound pressure but also the leading order terms. The series expansions derived for the propagation parameters extend the results given in the literature with additional terms resulting from viscosity and heat conduction in the core region. The second part is concerned with the nonlinear modulation of a wave packet transmitted through a real gas. A damped nonlinear Schrodinger equation is derived and its solutions for positive as well as negative values of the nonlinearity parameter are studied. In particular, the case of wave propagation in ducts containing a so-called BZT fluid is discussed. PMID- 15898628 TI - Simultaneous measurement of acoustic and streaming velocities in a standing wave using laser Doppler anemometry. AB - Laser Doppler anemometry (LDA) with burst spectrum analysis (BSA) is used to study the acoustic streaming generated in a cylindrical standing-wave resonator filled with air. The air column is driven sinusoidally at a frequency of approximately 310 Hz and the resultant acoustic-velocity amplitudes are less than 1.3 m/s at the velocity antinodes. The axial component of fluid velocity is measured along the resonator axis, across the diameter, and as a function of acoustic amplitude. The velocity signals are postprocessed using the Fourier averaging method [Sonnenberger et al., Exp. Fluids 28, 217-224 (2000)]. Equations are derived for determining the uncertainties in the resultant Fourier coefficients. The time-averaged velocity-signal components are seen to be contaminated by significant errors due to the LDA/BSA system. In order to avoid these errors, the Lagrangian streaming velocities are determined using the time harmonic signal components and the arrival times of the velocity samples. The observed Lagrangian streaming velocities are consistent with Rott's theory [N. Rott, Z. Angew. Math. Phys. 25, 417-421 (1974)], indicating that the dependence of viscosity on temperature is important. The onset of streaming is observed to occur within approximately 5 s after switching on the acoustic field. PMID- 15898629 TI - Influences of a temperature gradient and fluid inertia on acoustic streaming in a standing wave. AB - Following the experimental method of Thompson and Atchley [J. Acoust. Soc. Am. 117, 1828-1838 (2005)] laser Doppler anemometry (LDA) is used to investigate the influences of a thermoacoustically induced axial temperature gradient and of fluid inertia on the acoustic streaming generated in a cylindrical standing-wave resonator filled with air driven sinusoidally at a frequency of 308 Hz. The axial component of Lagrangian streaming velocity is measured along the resonator axis and across the diameter at acoustic-velocity amplitudes of 2.7, 4.3, 6.1, and 8.6 m/s at the velocity antinodes. The magnitude of the axial temperature gradient along the resonator wall is varied between approximately 0 and 8 K/m by repeating measurements with the resonator either surrounded by a water jacket, suspended within an air-filled tank, or wrapped in foam insulation. A significant correlation is observed between the temperature gradient and the behavior of the streaming: as the magnitude of the temperature gradient increases, the magnitude of the streaming decreases and the shape of the streaming cell becomes increasingly distorted. The observed steady-state streaming velocities are not in agreement with any available theory. PMID- 15898630 TI - On the interaction of counterpropagating acoustic waves in resonant rods composed of materials with hysteretic quadratic nonlinearity. AB - An ordinary inhomogeneous integrodifferential equation for the profile of the acoustic wave in a resonant rod, composed of a material with hysteretic quadratic nonlinearity, is derived. It explicitly takes into account the interaction of the counterpropagating acoustic waves. It also incorporates the boundary conditions in the sense that all possible solutions of the equation satisfy the appropriate boundary conditions at the ends of the rod. PMID- 15898631 TI - On the stability of the effective apodization of the nonlinearly generated second harmonic with respect to range. AB - The concept of an effective apodization was introduced to describe the field pattern for the nonlinearly generated second harmonic (2f) within the focal zone using a linear propagation model. Our objective in this study was to investigate the validity of the concept of an effective apodization at 2f as an approach to approximating the field of the second harmonic over a wide range of depths. Two experimental setups were employed: a vascular imaging array with a water path and an adult cardiac imaging array with an attenuating liver path. In both cases the spatial dependencies of the ultrasonic fields were mapped by scanning a point like hydrophone within a series of planes orthogonal to the propagation direction. The sampling distances were located before, within, and beyond the focal zone. The signals were Fourier transformed and the complex values at 2f were linearly backpropagated to the transmit plane in order to obtain an effective apodization. The measured results demonstrated a relatively constant effective apodization at 2f as a function of propagation distance. Finite amplitude computer simulations were found to be in agreement with these measurements. Thus the measure of the effective apodization at 2f provides an approximation to the second harmonic field outside the focal zone. PMID- 15898632 TI - Acoustic pulse propagation through a fluctuating stably stratified atmospheric boundary layer. AB - Mesoscale wind speed and temperature fluctuations with periods from 1 min to a few hours significantly affect temporal variability and turbulent regime of the stable atmospheric boundary layer (ABL). Their statistical characteristics are still poorly understood, although the knowledge of such statistics is required when modeling sound propagation through the stable ABL. Several field experiments have been conducted to study the influence of mesoscale wind speed fluctuations on acoustic pulse propagation through the stable ABL. Some results of these experiments are described in this paper. A special acoustic source was used to generate acoustic pulses by the detonation of an air-propane mixture with a repetition period 30 s. The mean wind speed and temperature profiles were continuously measured by Doppler sodar and temperature profiler, whereas mesoscale wind fluctuations were measured by anemometers placed on a 56-m mast. From the measurements of the pulse travel time fluctuations at different distances from the source, the statistical characteristics of the mesoscale wind fluctuations, such as frequency spectra, coherences, horizontal phase speeds and scales, have been obtained. Some of the obtained results are interpreted with the use of a recently developed model for the internal wave spectrum in a stably stratified atmosphere. PMID- 15898633 TI - The use of impedance matching capillaries for reducing resonance in rosette infrasonic spatial filters. AB - Rosette spatial filters are used at International Monitoring System infrasound array sites to reduce noise due to atmospheric turbulence. A rosette filter consists of several clusters, or rosettes, of low-impedance inlets. Acoustic energy entering each rosette of inlets is summed, acoustically, at a secondary summing manifold. Acoustic energy from the secondary manifolds are summed acoustically at a primary summing manifold before entering the microbarometer. Although rosette filters have been found to be effective at reducing infrasonic noise across a broad frequency band, resonance inside the filters reduces the effectiveness of the filters at high frequencies. This paper presents theoretical and observational evidence that the resonance inside these filters that is seen below 10 Hz is due to reflections occuring at impedance discontinuities at the primary and secondary summing manifolds. Resonance involving reflections at the inlets amplifies noise levels at frequencies above 10 Hz. This paper further reports results from theoretical and observational tests of impedance matching capillaries for removing the resonance problem. Almost total removal of resonant energy below 5 Hz was found by placing impedance matching capillaries adjacent to the secondary summing manifolds in the pipes leading to the primary summing manifold and the microbarometer. Theory and recorded data indicate that capillaries with resistance equal to the characteristic impedance of the pipe connecting the secondary and primary summing manifolds suppresses resonance but does not degrade the reception of acoustic signals. Capillaries at the inlets can be used to remove resonant energy at higher frequencies but are found to be less effective due to the high frequency of this energy outside the frequency band of interest. PMID- 15898634 TI - Simultaneous acoustic channel measurement via maximal-length-related sequences. AB - A wide variety of acoustic systems has multiple sources and receivers. This paper proposes a technique for making acoustic measurements simultaneously for multiple sources. The proposed technique features a collection of excitation signals of the maximum-length sequence (MLS) and MLS-related classes. Each signal in the set has a pulse-like autocorrelation function, and the cross-correlation functions between arbitrary pairs of signals drawn from the set have peak values that are significantly lower than the peak magnitude of the autocorrelation functions. The proposed method is particularly valuable when characterization of multisource, multireceiver system must be accomplished in a limited time period. Both simulation and experimental results are presented that demonstrate the feasibility and fidelity of the proposed techniques in characterizing acoustic systems. PMID- 15898635 TI - Phase speed and attenuation in bubbly liquids inferred from impedance measurements near the individual bubble resonance frequency. AB - In the ocean, natural and artificial processes generate clouds of bubbles that scatter and attenuate sound. Measurements have shown that at the individual bubble resonance frequency, sound propagation in this medium is highly attenuated and dispersive. The existing theory to explain this behavior is deemed adequate away from resonance. However, due to excessive attenuation near resonance, little experimental data exists for a comparison with model predictions. An impedance tube was developed specifically for exploring this regime. The effective medium phase speed and attenuation were inferred from measurements of the surface impedance of a layer of bubbly liquid composed of air bubbles and distilled water, for void fractions from 6.2 x 10(-5) to 5.4 x 10(-4) and bubble sizes centered around 0.62 mm in radius. Improved measurement speed, accuracy, and precision is possible with the new instrument, and both instantaneous and time averaged measurements were obtained. The phase speed and attenuation at resonance was observed to be sensitive to the bubble population statistics and agreed with an existing model [J. Acoust. Soc. Am. 85, 732-746 (1989)], within the uncertainty of the bubble population parameters. Agreement between the model and the data reported here is better than for the data that was available when the model was originally published. PMID- 15898636 TI - A phase-space Gaussian beam summation representation of rough surface scattering. AB - A Gaussian beam (GB) summation representation for rough surface scattering is introduced. In this scheme, the coherent and incoherent scattered fields are described by a phase-space summation of GBs that emanate from the rough surface at discrete set of points and directions. It thus involves stochastic GB2GB scattering matrices for the coherent and incoherent fields, and deterministic GB propagators. It benefits from the simplicity and accuracy of the latter, and can be used in applications involving propagation in complex scenarios comprising inhomogeneous media with rough surface boundaries. The GB2GB matrices are calculated from the statistical moments of the scattering amplitude, which are given either analytically or empirically. An analytical and numerical example for weakly rough surface is presented and discussed. Applications to the more complicated propagation scenario of doubly rough surface waveguide with multiple reflection phenomena will be presented in a follow-up publication. PMID- 15898637 TI - Phase-space beam summation analysis of rough surface waveguide. AB - A Gaussian beam summation (GBS) formulation is introduced for a doubly rough boundary waveguide, wherein the coherent and incoherent scattered fields are decomposed into a discrete phase-space summation of Gaussian beams (GB) that emanate from the rough surfaces in all directions. The scheme involves deterministic GB propagators and stochastic GB-to-GB (GB2GB) scattering matrices for the coherent and incoherent fields, where each scattered beam is propagated inside the waveguide and is scattered again from the rough boundaries. The GB2GB matrices are calculated from the statistical moments of the scattering amplitude, which are given either analytically or empirically. An analytical and numerical example for a waveguide with weak boundary roughness is presented and discussed. The formulation reveals explicitly the phase-space footprint of the stochastic multiple scattering process at the rough boundaries, thus providing a cogent physical interpretation and an effective mathematical representation to the field. The formulation also accommodates the receiver's pattern in the same phase space format. Bistatic reverberations inside a rough surface waveguide as a function of the range and of the source and the receiver directions are thus examined as an implementation example. PMID- 15898638 TI - Geoacoustic inversion in time domain using ship of opportunity noise recorded on a horizontal towed array. AB - A time domain geoacoustic inversion method using ship noise received on a towed horizontal array is presented. The received signal, containing ship noise as a source of opportunity, is time-reversed and then back-propagated to the vicinity of the ship. The back-propagated signal is correlated with the modeled signal which is expected to peak at the ship's location in case of a good match for the environment. This match is utilized for the geoacoustic parameter inversion. The objective function for this optimization problem is thus defined as the normalized power focused in an area around the source position, using a matched impulse response filter. A hybrid use of global and local search algorithms, i.e., GA and Powell's method is applied to the optimization problem. Applications of the proposed inversion method to MAPEX 2000 noise experiment conducted north of the island of Elba show promising results, and it is shown that the time domain inversion takes advantage of dominant frequencies of the source signature automatically. PMID- 15898639 TI - Effects of environmental uncertainties on sonar detection performance prediction. AB - The development of effective passive sonar systems depends upon the ability to accurately predict the performance of sonar detection algorithms in realistic ocean environments. Such environments are typically characterized by a high degree of uncertainty, thus limiting the usefulness of performance prediction approaches that assume a deterministic environment. Here we derive closed-form receiver operating characteristic (ROC) expressions for an optimal Bayesian detector and for several typical suboptimal detectors, based on a statistical model of environmental uncertainty. Various scenarios extended from an NRL benchmark shallow-water model were used to check the analytical ROC expressions and to illustrate the effect of environmental uncertainty on detection performance. The results showed that (1) optimal detection performance in an uncertain environment in diffuse noise depends primarily on the signal-to-noise ratio at the receivers and the rank of the signal matrix, where the rank is an effective representation of the scale of environmental uncertainty; (2) the ROC expression for the optimal Bayesian detector provides a more realistic performance upper bound than that obtained from conventional sonar equations that do not incorporate environmental uncertainty; and (3) detection performance predictions can be performed much faster than with commonly used numerical methods such as Monte Carlo performance evaluations. PMID- 15898640 TI - Bayesian sonar detection performance prediction in the presence of interference in uncertain environments. AB - The detection performance of sonar systems can be greatly limited by the presence of interference and environmental uncertainty. The classic sonar equation does not take into account these two limiting factors and is inaccurate in predicting sonar detection performance. Here we have developed closed-form receiver operating characteristic (ROC) performance expressions for the Bayesian detector in the presence of interference in uncertain environments. Various scenarios extended from a NRL benchmark shallow-water model were used to test the analytical ROC expressions and to analyze the effects of interference and environmental uncertainty on detection performance. The results show that (1) the degradation on detection performance due to interference is greatly magnified by the presence of environmental uncertainty; (2) Bayesian sonar detection performance depends on the following fundamental parameters: the signal-to-noise ratio, the rank of the signal matrix, and the signal-to-interference coefficient; (3) the proposed analytical ROC performance predictions can be computed much faster than performance evaluations with commonly used Monte Carlo techniques. PMID- 15898641 TI - Concurrent inversion of geo- and bio-acoustic parameters from transmission loss measurements in the Yellow Sea. AB - This paper describes results of a simultaneous inversion of bio-acoustic parameters of fish (anchovies) and geo-acoustic parameters of the bottom from transmission loss (TL) measurements in the Yellow Sea, which were reported by Qiu et al. [J. Sound Vib. 220, 331-342 (1999)]. This data set was selected because the bio-absorptivity at their site was extremely large, 40 dB at 1.3 kHz at 5 km, and measurements were made between multiple source and receiver depths and ranges. Measurements were made at night when anchovies are generally dispersed. Replica fields were calculated with a normal mode model, which incorporates bio absorption layers. The inversion was based on minimizing the rms difference, delta, between measured and calculated values of TL at all ranges and source and receiver depths, and involved a simultaneous search for bio-layer depth, bio layer thickness, bio-alpha, geo-sound speed, and geo-alpha. The resultant small value of delta, +/- 1.7 dB, confirmed that the model, which was assumed in replica field calculations, was realistic, and that inverted parameters were meaningful. In particular, the inverted depth of the bio-absorption layer, 6.9 +/ 0.3 m, was consistent with theoretical calculations of the depth, 5.8 +/- 1 m, of 10-cm-long anchovies Engraulis japonicus, the dominant species in the Yellow Sea. PMID- 15898642 TI - Long range acoustic imaging of the continental shelf environment: the Acoustic Clutter Reconnaissance Experiment 2001. AB - An active sonar system is used to image wide areas of the continental shelf environment by long-range echo sounding at low frequency. The bistatic system, deployed in the STRATAFORM area south of Long Island in April-May of 2001, imaged a large number of prominent clutter events over ranges spanning tens of kilometers in near real time. Roughly 3000 waveforms were transmitted into the water column. Wide-area acoustic images of the ocean environment were generated in near real time for each transmission. Between roughly 10 to more than 100 discrete and localized scatterers were registered for each image. This amounts to a total of at least 30000 scattering events that could be confused with those from submerged vehicles over the period of the experiment. Bathymetric relief in the STRATAFORM area is extremely benign, with slopes typically less than 0.5 degrees according to high resolution (30 m sampled) bathymetric data. Most of the clutter occurs in regions where the bathymetry is locally level and does not coregister with seafloor features. No statistically significant difference is found in the frequency of occurrence per unit area of repeatable clutter inside versus outside of areas occupied by subsurface river channels. PMID- 15898643 TI - Time-reversal imaging for classification of submerged elastic targets via Gibbs sampling and the Relevance Vector Machine. AB - Time-reversal imaging (TRI) is analogous to matched-field processing, although TRI is typically very wideband and is appropriate for subsequent target classification (in addition to localization). Time-reversal techniques, as applied to acoustic target classification, are highly sensitive to channel mismatch. Hence, it is crucial to estimate the channel parameters before time reversal imaging is performed. The channel-parameter statistics are estimated here by applying a geoacoustic inversion technique based on Gibbs sampling. The maximum a posteriori (MAP) estimate of the channel parameters are then used to perform time-reversal imaging. Time-reversal implementation requires a fast forward model, implemented here by a normal-mode framework. In addition to imaging, extraction of features from the time-reversed images is explored, with these applied to subsequent target classification. The classification of time reversed signatures is performed by the relevance vector machine (RVM). The efficacy of the technique is analyzed on simulated in-channel data generated by a free-field finite element method (FEM) code, in conjunction with a channel propagation model, wherein the final classification performance is demonstrated to be relatively insensitive to the associated channel parameters. The underlying theory of Gibbs sampling and TRI are presented along with the feature extraction and target classification via the RVM. PMID- 15898644 TI - Protocols for calibrating multibeam sonar. AB - Development of protocols for calibrating multibeam sonar by means of the standard target method is documented. Particular systems used in the development work included three that provide the water-column signals, namely the SIMRAD SM2000/90 and 200-kHz sonars and RESON SeaBat 8101 sonar, with operating frequency of 240 kHz. Two facilities were instrumented specifically for the work: a sea well at the Woods Hole Oceanographic Institution and a large, indoor freshwater tank at the University of New Hampshire. Methods for measuring the transfer characteristics of each sonar, with transducers attached, are described and illustrated with measurement results. The principal results, however, are the protocols themselves. These are elaborated for positioning the target, choosing the receiver gain function, quantifying the system stability, mapping the directionality in the plane of the receiving array and in the plane normal to the central axis, measuring the directionality of individual beams, and measuring the nearfield response. General preparations for calibrating multibeam sonars and a method for measuring the receiver response electronically are outlined. Advantages of multibeam sonar calibration and outstanding problems, such as that of validation of the performance of multibeam sonars as configured for use, are mentioned. PMID- 15898645 TI - Multi-mode Lamb wave tomography with arrival time sorting. AB - Lamb wave tomography has been shown to be an effective nondestructive evaluation technique for platelike structures. A series of pitch-catch measurements between ultrasonic transducers can be taken from different orientations across the sample to create a map of a particular feature of interest such as plate thickness. Most previous work has relied solely on the first arriving mode for the time-of-flight measurements and tomographic reconstructions. The work described here demonstrates the capability of the Lamb wave tomography system to generate accurate reconstructions from multiple modes. Because each mode has different through-thickness displacement values, each is sensitive to different types of flaws, and the information gained from a multi-mode analysis can improve understanding of the structural integrity of the inspected material. However, one of the problems with the extraction of multi-mode arrival times is that destructive interference between two modes may cause one of the modes to seemingly disappear in the signal. The goal of the sorting algorithm presented in this work is to try and counteract this problem by using multiple frequency scans -also known as frequency walking--to sort the arrival times into their correct mode series. PMID- 15898646 TI - Regularization method for measurement of structural intensity using nearfield acoustical holography. AB - The regularization method for measurement of structural intensity using nearfield acoustical holography is proposed. Spatial derivatives of normal displacement are necessary to obtain the structural intensity. The derivative operations amplify high-wave-number components of measurement noise. Therefore, the estimation of an appropriate wave-number filter is crucial for implementation of the measurement of structural intensity. In conventional methods, this wave-number filter is determined from the flexural wavelength. And the same wave-number filter is applied to obtain all spatial derivatives. As a result, structural intensity obtained from the pressure hologram, whose signal-to-noise ratio is low, is seriously contaminated by the noise. To overcome this difficulty, regularization theory is applied to determine the appropriate wave-number filter for each order of derivatives. The effectiveness of the proposed method is demonstrated by experiments. PMID- 15898647 TI - Smart panels with velocity feedback control systems using triangularly shaped strain actuators. AB - In this paper we present a theoretical study on the active structural acoustic control of a new smart panel with sixteen triangularly shaped piezoelectric patch actuators, having their base edges evenly distributed along the perimeter of the panel, and velocity sensors positioned at the vertices opposite the base edges. The performance is assessed and contrasted with that of a conventional smart panel using a 4 x 4 array of square piezoelectric patch actuators evenly distributed over the surface of the panel with velocity sensors at their centers. For both systems the control effectiveness and stability of MIMO decentralized or SISO direct velocity feedback control architectures have been analyzed. The two control systems are arranged to generate active damping which reduces the response and sound radiation of the panel in the lightly damped and well separated low-frequency resonances. In particular the new control system can be seen as a set of sixteen "active wedges" which absorb energy from the incident flexural waves to the borders of the panel so that the panel could be considered anechoic. This study shows that the new arrangement with triangularly shaped actuators can achieve better control than the corresponding system using square actuators. PMID- 15898648 TI - Reconstruction of transient acoustic radiation from a sphere. AB - Transient near-field acoustical holography (NAH) formulation is derived from the Helmholtz equation least squares (HELS) method to reconstruct acoustic radiation from a spherical surface subject to transient excitations in a free field. To facilitate derivations of temporal solutions, we make use of the Laplace transform and expansion in terms of the spherical Hankel functions and spherical harmonics, with their coefficients settled by solving a system of equations obtained by matching an assumed-form solution to the measured acoustic pressure. To derive a general form of solution for a temporal kernel, we replace the spherical Hankel functions and their derivatives by polynomials, recast infinite integrals in the inverse Laplace transform as contour integrals in a complex s plane, and evaluate it via the residue theorem. The transient acoustic quantities anywhere including the source surface are then obtained by convoluting the temporal kernels with respect to the measured acoustic pressure. Numerical examples of reconstructing transient acoustic fields from explosively expanding, impulsively accelerating, and partially accelerating spheres, and that from a sphere subject to an arbitrarily time-dependent excitation are depicted. To illustrate the effectiveness of HELS-based transient NAH formulations, all input data are collected along an arbitrarily selected line segment and used to reconstruct transient acoustic quantities everywhere. PMID- 15898649 TI - Analytical approach for sound attenuation in perforated dissipative silencers with inlet/outlet extensions. AB - The acoustic attenuation performance of perforated dissipative circular expansion chambers with inlet/outlet extensions is investigated. The eigenvalues and eigenfunctions of the sound field are analytically determined in the extended inlet/outlet circular ducts, upstream/downstream end annular dissipative chambers, and the central perforated dissipative expansion chamber. Utilizing the continuity conditions of velocity/pressure at the interfaces the transmission loss is predicted by a two-dimensional analytical approach. For a specific configuration, such predictions are compared with both experiments and a three dimensional computational solution based on the substructure boundary element technique, showing a reasonable agreement. The analytical results for the effect of the absorbent resistivity, duct porosity, and geometryon the acoustic attenuation performance are discussed in detail. PMID- 15898650 TI - Absorptive properties of rigid porous media: application to face centered cubic sphere packing. AB - The classical description of porous media as a homogeneous equivalent fluid is presented, and its foundations on the homogenization method is introduced and applied to the numerical prediction model for a periodic porous medium composed by face centered cubic sphere packing on which measurements have been made. The results are compared with existing numerical results in the literature and with new and experimental data. PMID- 15898651 TI - Theory and design of sound field reproduction in reverberant rooms. AB - With the recent emergence of surround sound technology, renewed interest has been shown in the problem of sound field reproduction. However, in practical acoustical environments, the performance of sound reproduction techniques are significantly degraded by reverberation. In this paper, we develop a method of sound field reproduction for reverberant environments. The key to this method is an efficient parametrization of the acoustic transfer function over a region of space. Using this parametrization, a practical method has been provided for determining the transfer function between each loudspeaker and every point in the reproduction region. Through several simulation examples, the reverberant field designs have been shown to yield a reproduction accuracy as good as conventional free-field designs, and better than multipoint least squares designs when loudspeaker numbers are limited. The successful reproduction of sound over a wide frequency range has also been demonstrated. This approach reveals the appropriate choices for fundamental design parameters. PMID- 15898652 TI - Microphone array signal processing with application in three-dimensional spatial hearing. AB - Microphone arrays are known to enhance the directionality and signal-to-noise ratio (SNR) over single-channel sensors. This is considered beneficial in many applications such as video-conferencing systems and hearing aids. However, this advantage comes at the price of the sensation of spatial hearing. The spatial cues due to diffractions of the head and torso are lost if the array is not fitted in the ears. In this paper we present a system that incorporates binaural hearing synthesis into array signal processing, in an attempt to recover the three-dimensional sound image that a human listener would naturally perceive. In the system, the superdirective beamformer is exploited to estimate the direction of arrival (DOA) of the incoming sound. The spatial sound image is restored by steering the beam to the direction found in the DOA session and filtering the array output with the corresponding Head Related Transfer Functions (HRTF). The algorithms have been implemented in real-time fashion using a digital signal processor. Objective and subjective experiments were performed to validate the proposed system. The experimental results showed that the accurate localization of the sound source is achievable using the array system. PMID- 15898653 TI - The shift-invariant discrete wavelet transform and application to speech waveform analysis. AB - The discrete wavelet transform may be used as a signal-processing tool for visualization and analysis of nonstationary, time-sampled waveforms. The highly desirable property of shift invariance can be obtained at the cost of a moderate increase in computational complexity, and accepting a least-squares inverse (pseudoinverse) in place of a true inverse. A new algorithm for the pseudoinverse of the shift-invariant transform that is easier to implement in array-oriented scripting languages than existing algorithms is presented together with self contained proofs. Representing only one of the many and varied potential applications, a recorded speech waveform illustrates the benefits of shift invariance with pseudoinvertibility. Visualization shows the glottal modulation of vowel formants and frication noise, revealing secondary glottal pulses and other waveform irregularities. Additionally, performing sound waveform editing operations (i.e., cutting and pasting sections) on the shift-invariant wavelet representation automatically produces quiet, click-free section boundaries in the resulting sound. The capabilities of this wavelet-domain editing technique are demonstrated by changing the rate of a recorded spoken word. Individual pitch periods are repeated to obtain a half-speed result, and alternate individual pitch periods are removed to obtain a double-speed result. The original pitch and formant frequencies are preserved. In informal listening tests, the results are clear and understandable. PMID- 15898654 TI - Defect imaging with guided waves in a pipe. AB - Guided wave techniques are expected to become an effective means for rapid, long range inspection of pipes. Such techniques still have many practical difficulties in application, however, due to the complex characteristics of guided waves such as dispersion and their multimodal nature. A defect imaging technique is developed in this study to overcome the complexities of guided wave inspection. Received signals are separated into single-mode waveforms with a mode extraction technique and then spatial waveforms on the pipe surface at an arbitrary time are reconstructed. The predicted waveforms can provide a defect image at the moment when an incident wave arrives at a defect region, which is based on a time reversal technique. This defect imaging technique is experimentally verified using eight signals detected at eight different circumferential positions. Images of artificial defects are obtained with one-hole and two-hole test pipes, and increasing the frequency of incident waves increases the resolution of the images. Holes and pipe ends are recognizable in the images, but the reconstructed images contain some errors in the area behind the defects where guided waves do not propagate or do not reflect back to the receiving transducers. PMID- 15898655 TI - Plified nonlinear outer hair cell models. AB - We present a consistent second-order expansion of nonlinear constitutive theories for outer hair cells. For a particular theory, we will test the validity of such a model for small variations in voltage and strain about the resting state of outer hair cells. An analysis of the various terms in the simplified nonlinear model and their relevance to outer hair cell mechanics are presented. Results show that the second-order expansion is adequate for modeling outer hair cell mechanics in a global model of the cochlea. Model predictions agree with the notion that voltage nonlinearities are the dominant ones at low sound levels in vivo. PMID- 15898656 TI - Modeling high-frequency electromotility of cochlear outer hair cell in microchamber experiment. AB - Cochlear outer hair cells (OHC) are critically important for the amplification and sharp frequency selectivity of the mammalian ear. The microchamber experiment has been an effective tool to analyze the OHC high-frequency performance. In this study, the OHC electrical stimulation in the microchamber is simulated. The model takes into account the inertial and viscous properties of fluids inside and outside the cell as well as the viscoelastic and piezoelectric properties of the cell composite membrane (wall). The closed ends of the cylindrical cell were considered as oscillatory rigid plates. The final solution was obtained in terms of Fourier series, and it was checked against the available results of the microchamber experiment. The conditions of the interaction between the cell and pipette was analyzed, and it was found that the amount of slip along the contact surface has a significant effect on the cell electromotile response. The cell's length changes were computed as a function of frequency, and their dependence on the viscosities of both fluids and the cell wall was analyzed. The distribution of the viscous losses inside the fluids was also estimated. The proposed approach can help in a better understanding of the high-frequency OHC electromotility under experimental and physiological conditions. PMID- 15898657 TI - Modulation masking produced by second-order modulators. AB - Recent studies suggest that an auditory nonlinearity converts second-order sinusoidal amplitude modulation (SAM) (i.e., modulation of SAM depth) into a first-order SAM component, which contributes to the perception of second-order SAM. However, conversion may also occur in other ways such as cochlear filtering. The present experiments explored the source of the first-order SAM component by investigating the ability to detect a 5-Hz, first-order SAM probe in the presence of a second-order SAM masker beating at the probe frequency. Detection performance was measured as a function of masker-carrier modulation frequency, phase relationship between the probe and masker modulator, and probe modulation depth. In experiment 1, the carrier was a 5-kHz sinusoid presented either alone or within a notched-noise masker in order to restrict off-frequency listening. In experiment 2, the carrier was a white noise. The data obtained in both carrier conditions are consistent with the existence of a modulation distortion component. However, the phase yielding poorest detection performance varied across experimental conditions between 0 degrees and 180 degrees, confirming that, in addition to nonlinear mechanisms, cochlear filtering and off-frequency listening play a role in second-order SAM perception. The estimated magnitude of the modulation distortion component ranges from 5%-12%. PMID- 15898658 TI - The effect of spatial separation on informational masking of speech in normal hearing and hearing-impaired listeners. AB - The ability to understand speech in a multi-source environment containing informational masking may depend on the perceptual arrangement of signal and masker objects in space. In normal-hearing listeners, Arbogast et al. [J. Acoust. Soc. Am. 112, 2086-2098 (2002)] found an 18-dB spatial release from a primarily informational masker, compared to 7 dB for a primarily energetic masker. This article extends the earlier work to include the study of listeners with sensorineural hearing loss. Listeners performed closed-set speech recognition in two spatial conditions: 0 degrees and 90 degrees separation between signal and masker. Three maskers were tested: (1) the different-band sentence masker was designed to be primarily informational; (2) the different-band noise masker was a control for the different-band sentence; and (3) the same-band noise masker was designed to be primarily energetic. The spatial release from the different-band sentence was larger than for the other maskers, but was smaller (10 dB) for the hearing-impaired group than for the normal-hearing group (15 dB). The smaller benefit for the hearing-impaired listeners can be partially explained by masker sensation level. However, the results suggest that hearing-impaired listeners can use the perceptual effect of spatial separation to improve speech recognition in the presence of a primarily informational masker. PMID- 15898659 TI - A Speech Intelligibility Index-based approach to predict the speech reception threshold for sentences in fluctuating noise for normal-hearing listeners. AB - The SII model in its present form (ANSI S3.5-1997, American National Standards Institute, New York) can accurately describe intelligibility for speech in stationary noise but fails to do so for nonstationary noise maskers. Here, an extension to the SII model is proposed with the aim to predict the speech intelligibility in both stationary and fluctuating noise. The basic principle of the present approach is that both speech and noise signal are partitioned into small time frames. Within each time frame the conventional SII is determined, yielding the speech information available to the listener at that time frame. Next, the SII values of these time frames are averaged, resulting in the SII for that particular condition. Using speech reception threshold (SRT) data from the literature, the extension to the present SII model can give a good account for SRTs in stationary noise, fluctuating speech noise, interrupted noise, and multiple-talker noise. The predictions for sinusoidally intensity modulated (SIM) noise and real speech or speech-like maskers are better than with the original SII model, but are still not accurate. For the latter type of maskers, informational masking may play a role. PMID- 15898660 TI - Perception of pitch location within a speaker's F0 range. AB - Fundamental frequency (F0) is used for many purposes in speech, but its linguistic significance is based on its relation to the speaker's range, not its absolute value. While it may be that listeners can gauge a specific pitch relative to a speaker's range by recognizing it from experience, whether they can do the same for an unfamiliar voice is an open question. The present experiment explored that question. Twenty native speakers of English (10 male, 10 female) produced the vowel /a/ with a spoken (not sung) voice quality at varying pitches within their own ranges. Listeners then judged, without familiarization or context, where each isolated F0 lay within each speaker's range. Correlations were high both for the entire range (0.721) and for the range minus the extremes (0.609). Correlations were somewhat higher when the F0s were related to the range of all the speakers, either separated by sex (0.830) or pooled (0.848), but several factors discussed here may help account for this pattern. Regardless, the present data provide strong support for the hypothesis that listeners are able to locate an F0 reliably within a range without external context or prior exposure to a speaker's voice. PMID- 15898661 TI - Perception of aperiodicity in pathological voice. AB - Although jitter, shimmer, and noise acoustically characterize all voice signals, their perceptual importance in naturally produced pathological voices has not been established psychoacoustically. To determine the role of these attributes in the perception of vocal quality, listeners were asked to adjust levels of jitter, shimmer, and the noise-to-signal ratio in a speech synthesizer, so that synthetic voices matched naturally produced tokens. Results showed that, although listeners agreed well in their judgments of the noise-to-signal ratio, they did not agree with one another in their chosen settings for jitter and shimmer. Noise-dependent differences in listeners' ability to detect changes in amounts of jitter and shimmer implicate both listener insensitivity and inability to isolate jitter and shimmer as separate dimensions in the overall pattern of aperiodicity in a voice as causes of this poor agreement. These results suggest that jitter and shimmer are not useful as independent indices of perceived vocal quality, apart from their acoustic contributions to the overall pattern of spectrally shaped noise in a voice. PMID- 15898662 TI - Consonant recognition and the articulation index. AB - The purpose of this paper is to provide insight into how speech is processed by the auditory system, by quantifying the nature of nonsense speech sound confusions. (1) The Miller and Nicely [J. Acoust. Soc. Am. 27(2), 338-352 (1955)] confusion matrix (CM) data are analyzed by plotting the CM elements Si,j(SNR) as a function of the signal-to-noise ratio (SNR). This allows for the robust clustering of perceptual feature (event) groups, not robustly defined by a single CM table, where clusters depend on the sound order. (2) The SNR is then re expressed as an articulation index (AI), and used as the independent variable. The normalized log scores log(1-Si,i(AI)) and log(Si,j(AI)), j not equal to i, then become linear functions of AI, on log-error versus AI plots. This linear dependence may be interpreted as an extension of the band-independence model of Fletcher. (3) The model formula for the average score for the finite-alphabet case Pc(AI,H)= sigma(N)i=1Si,i/N is then modified to include the effect of entropy H. Due to the grouping of sounds with increased SNR (and AI), the sound group entropy Hg plays a key role in this performance measure. (4) A parametric model for the confusions Si,j(AI,Hg) is then described, which characterizes the confusions between competing sounds within a group. PMID- 15898663 TI - Coherence and the speech intelligibility index. AB - The speech intelligibility index (SII) (ANSI S3.5-1997) provides a means for estimating speech intelligibility under conditions of additive stationary noise or bandwidth reduction. The SII concept for estimating intelligibility is extended in this paper to include broadband peak-clipping and center-clipping distortion, with the coherence between the input and output signals used to estimate the noise and distortion effects. The speech intelligibility predictions using the new procedure are compared with intelligibility scores obtained from normal-hearing and hearing-impaired subjects for conditions of additive noise and peak-clipping and center-clipping distortion. The most effective procedure divides the speech signal into low-, mid-, and high-level regions, computes the coherence SII separately for the signal segments in each region, and then estimates intelligibility from a weighted combination of the three coherence SII values. PMID- 15898664 TI - Statistical properties of infant-directed versus adult-directed speech: insights from speech recognition. AB - Previous studies have shown that infant-directed speech ('motherese') exhibits overemphasized acoustic properties which may facilitate the acquisition of phonetic categories by infant learners. It has been suggested that the use of infant-directed data for training automatic speech recognition systems might also enhance the automatic learning and discrimination of phonetic categories. This study investigates the properties of infant-directed vs. adult-directed speech from the point of view of the statistical pattern recognition paradigm underlying automatic speech recognition. Isolated-word speech recognizers were trained on adult-directed vs. infant-directed data sets and were tested on both matched and mismatched data. Results show that recognizers trained on infant-directed speech did not always exhibit better recognition performance; however, their relative loss in performance on mismatched data was significantly less severe than that of recognizers trained on adult-directed speech and presented with infant-directed test data. An analysis of the statistical distributions of a subset of phonetic classes in both data sets showed that this pattern is caused by larger class overlaps in infant-directed speech. This finding has implications for both automatic speech recognition and theories of infant speech perception. PMID- 15898665 TI - The acoustics of Japanese wooden drums called "mokugyo". AB - A drumlike traditional Japanese instrument, the mokugyo, is experimentally discussed. First, the acoustic characteristics of 176 mokugyos with diameters ranging from 7.5 to 120 cm and three drumsticks were measured. Results show that (a) the sound spectra consist of two common peaks [F1 (Hz): first peak frequency, F2 (Hz): second peak frequency] close together, with an average ratio (F2/F1) of 1.15, and (b) a drumstick beating the mokugyo is translated into an impact force applied over a period of time from 1 to 6 ms related to the mass and stiffness of the material wrapped around the tip of the drumstick. Second, to evaluate the acoustic response of a mokugyo in the final tuning process, the mechanical and acoustical analogy between the mokugyo and a bass reflex loudspeaker is theoretically and experimentally discussed. Results show that the model can be estimated within a relative error of 0.52% from the mass of wood chips. Finally, from a psychological experiment, the timbre of the mokugyo shows higher scores on psychological scales' when the ratio (F2/F1) becomes 1.15. PMID- 15898666 TI - Beating frequency and amplitude modulation of the piano tone due to coupling of tones. AB - The influence on a piano tone from weak coexcitation of damped adjacent tones due to coupling via the bridge is studied. The frequency and amplitude modulation of the sound resulting from coexcitation of one strong and one or two weak tones is analyzed. One weak tone causes frequency and amplitude modulation of the sound, and two weak tones produce beating frequency and amplitude modulation, where the beatings of the two modulations are of opposite phase. By digital recording of the sound of piano tones, the appearance of these phenomena is verified. The audibility of the observed frequency and amplitude modulation is discussed in terms of previously determined detection thresholds. The beating character of both frequency and amplitude modulations, however, distinguishes the phenomena from those previously studied and prompts further psychoacoustic investigations. It is shown that detuning of unison strings may significantly increase the frequency deviation of the frequency modulation in conjunction with affected amplitude modulation. The modulatory effects of coupling to adjacent tones therefore may possibly be utilized in the tuning process. A coupling of tones analogous to the situation in a piano may arise in other stringed musical instruments transferring string vibrations to a soundboard via a bridge. PMID- 15898667 TI - Generation of longitudinal vibrations in piano strings: from physics to sound synthesis. AB - Longitudinal vibration of piano strings greatly contributes to the distinctive character of low piano notes. In this paper a simplified modal model is developed, which describes the generation of phantom partials and longitudinal free modes jointly. The model is based on the simplification that the coupling from the transverse vibration to the longitudinal polarization is unidirectional. The modal formulation makes it possible to predict the prominent components of longitudinal vibration as a function of transverse modal frequencies. This provides a qualitative insight into the generation of longitudinal vibration, while the model is still capable of explaining the empirical results of earlier works. The semi-quantitative agreement with measurement results implies that the main source of phantom partials is the transverse to longitudinal coupling, while the string termination and the longitudinal to transverse coupling have only small influence. The results suggest that the longitudinal component of the tone can be treated as a quasi-harmonic spectrum with formantlike peaks at the longitudinal modal frequencies. The model is further simplified and applied for the real-time synthesis of piano sound with convincing sonic results. PMID- 15898668 TI - Aerodynamic excitation and sound production of blown-closed free reeds without acoustic coupling: the example of the accordion reed. AB - The accordion reed is an example of a blown-closed free reed. Unlike most oscillating valves in wind musical instruments, self-sustained oscillations occur without acoustic coupling. Flow visualizations and measurements in water show that the flow can be supposed incompressible and potential. A model is developed and the solution is calculated in the time domain. The excitation force is found to be associated with the inertial load of the unsteady flow through the reed gaps. Inertial effect leads to velocity fluctuations in the reed opening and then to an unsteady Bernoulli force. A pressure component generated by the local reciprocal air movement around the reed is added to the modeled aerodynamic excitation pressure. Since the model is two-dimensional, only qualitative comparisons with air flow measurements are possible. The agreement between the simulated pressure waveforms and measured pressure in the very near-field of the reed is reasonable. In addition, an aeroacoustic model using the permeable Ffowcs Williams-Hawkings integral method is presented. The integral expressions of the far-field acoustic pressure are also computed in the time domain. In agreement with experimental data, the sound is found to be dominated by the dipolar source associated by the strong momentum fluctuations of the flow through the reed gaps. PMID- 15898669 TI - Measurement of the depth-dependent resonance of water-loaded human lungs. AB - An experiment was conducted to determine the response of the human lung to water borne sound in the range of 20 to 500 Hz. A small pool inside a hyperbaric chamber was used to simulate four ambient pressure conditions spanning the range of recreational diving depths. Ten subjects were tested on two occasions each using three separate measures to evaluate the response of the subjects' lungs. With some notable exceptions, results were consistent between subjects and between measures. These indicate that human lungs can be reasonably modeled as a lumped single-degree-of-freedom system over the lower portion of the band of interest. Here, the surrounding fluid provides the dominant mass and the dominant stiffness is provided by the entrapped air with a small additional contribution from tissue elasticity. Measured resonances increase with the square root of ambient pressure from an average of 40 Hz with a quality factor of 1.8 at near surface pressure to 73 Hz with a quality factor of 2.6 at an equivalent depth of 36.4 m. There is evidence of other resonances within or near the band of interest that may be attributable to nonvolumetric chest/lung modes, Helmholtz resonance, and/or resonance of gastrointestinal bubbles. PMID- 15898670 TI - Instrumenting free-swimming dolphins echolocating in open water. AB - Dolphins within the Navy Marine Mammal Program use echolocation to effectively locate underwater mines. They currently outperform manmade systems at similar tasks, particularly in cluttered environments and on buried targets. In hopes of improving manmade mine-hunting sonar systems, two instrumentation packages were developed to monitor free-swimming dolphin motion and echolocation during open water target detection tasks. The biosonar measurement tool (BMT) is carried by a dolphin and monitors underwater position and attitude while simultaneously recording echolocation clicks and returning echoes through high-gain binaural receivers. The instrumented mine simulator (IMS) is a modified bottom target that monitors echolocation signals arriving at the target during ensonification. Dolphin subjects were trained to carry the BMT in open-bay bottom-object target searches in which the IMS could serve as a bottom object. The instrumentation provides detailed data that reveal hereto-unavailable information on the search strategies of free-swimming dolphins conducting open-water, bottom-object search tasks with echolocation. PMID- 15898671 TI - Echolocation characteristics of free-swimming bottlenose dolphins during object detection and identification. AB - A biosonar measurement tool (BMT) was created to investigate dolphin echolocation search strategies by recording echolocation clicks, returning echoes, and three dimensional angular motion, velocity, and depth of free-swimming dolphins performing open-water target detections. Trial start and stop times, locations determined from a differential global positioning system (DGPS), and BMT motion and acoustic data were used to produce spatial and acoustic representations of the searches. Two dolphins (LUT, FLP) searched for targets lying on the seafloor of a bay environment while carrying the BMT. LUT searched rapidly (< 10 s), produced few clicks, and varied click-peak frequency (20-120 kHz); FLP searched relatively slowly (tens of seconds) and produced many hundreds of clicks with stereotypical frequency-dependent energy distributions dominating from 30-60 kHz. Dolphins amplified target echo returns by either increasing the click source level or reducing distance to the target but without reducing source level. The distribution of echolocation click-peak frequencies suggested a bias in the dominant frequency components of clicks, possibly due to mechanical constraints of the click generator. Prior training and hearing loss accommodation potentially explain differences in the search strategies of the two dolphins. PMID- 15898672 TI - Quantifying the acoustic repertoire of a population: the vocalizations of free ranging bottlenose dolphins in Fiordland, New Zealand. AB - Quantification of the vocal repertoire of a species is critical for subsequent analysis of signal functionality, geographical variation, social relevance, and transmission. While signal repertoires have been documented for numerous animal species, detailed descriptions for the cetaceans are rare. This study describes the vocalizations of free-ranging bottlenose dolphins resident to the waters of Fiordland, New Zealand. The field recordings presented in this paper were made using both audio band and broadband apparatus. Subsequent classification of vocalizations using multivariate parameters enabled a quantification of the entire vocal output of the focal animals. These results were used to propose a meaningful repertoire of signals employed by this species. A total of 12 individual signal types were described, comprising four broad structural classes: "tonal," "single bursts," "click bursts," and "repeat bursts." The proposed repertoire will allow subsequent investigation into vocal behavior. It appears that the successful description of a species' repertoire is dependent on the use of appropriate recording systems, a high number of representative recordings with good signal-to-noise ratio, and subsequent validation of the original classification system. PMID- 15898673 TI - Information theory analysis of patterns of modulation in the advertisement call of the male bullfrog, Rana catesbeiana. AB - Male bullfrogs often amplitude modulate the envelopes of the individual notes (croaks) in their multinote advertisement calls. These amplitude modulations change the envelope of the note from smooth and unmodulated to one with varying numbers of modulations. A Markov analysis shows the pattern of change in the envelope to be highly ordered, but not completely so (semi-Markovian). Three simple rules govern the presence or absence of modulations in individual notes. These rules are (1) all calls begin with an unmodulated note; (2) the first note to be modulated will contain only one modulation; and (3) when a change in modulation occurs from one note to the next, it does so with an increase or a decrease of one modulation only. The addition of modulations is correlated with an increase in note duration. Physiologically, the presence of modulations might increase the precision of temporal coding of note periodicities in the central auditory system. PMID- 15898674 TI - Gas bubble and solid sphere motion in elastic media in response to acoustic radiation force. AB - The general approach to estimate the displacement of rounded objects (specifically, gas bubbles and solid spheres) in elastic incompressible media in response to applied acoustic radiation force is presented. In this study, both static displacement and transient motion are analyzed using the linear approximation. To evaluate the static displacement of the spherical inclusion, equations coupling the applied force, displacement, and shear modulus of the elastic medium are derived. Analytical expressions to estimate the static displacement of solid spheres and gas bubbles are presented. Under a continuously applied static force, both the solid sphere and the initially spherical gas bubble are displaced, and the bubble is deformed. The transient responses of the inclusions are described using motion equations. The displacements of the inclusion in elastic incompressible lossless media are analyzed using both frequency-domain and time-domain formalism, and the equations of motion are derived for both a solid sphere and a gas bubble. For a short pulsed force, an analytical solution for the equations of motion is presented. Finally, transient displacement of the gas bubble in viscoelastic media is considered. PMID- 15898676 TI - [Stand up, colleagues!]. PMID- 15898675 TI - The mechanical and thermal effects of focused ultrasound in a model biological material. AB - This paper is motivated by possible medical applications of focused ultrasound in the minimally invasive treatment of a variety of musculoskeletal disorders that are responsive to thermal treatment. A model-based analysis of the interaction of high-intensity focused ultrasound with biological materials is carried out in an effort to predict the path of the sound waves and the temperature field in the focal region. A finite-element-based general purpose code called PZFlex is used to determine the effects of nonlinearity and geometrical complexity of biological structures. It was found that at frequencies of interest in therapeutic applications, the nonlinear effects are usually negligible and the geometrical complexities can be handled through a substructuring procedure. An approximate analytical method with acceptable accuracy is developed as an alternative to the purely numerical approach used in PZFlex. The mechanical and thermal effects in two-layered fluid material systems induced by high-frequency focused ultrasound are calculated through this analytical method. The results are compared with those obtained using PZFlex as a benchmark. PMID- 15898677 TI - [Treating hypertension: a challenge for the practioner]. AB - Hypertension is associated with a greater risk of developing a stroke or a coronary heart disease, but remains, for the meanwhile, undertreated in a vast majority of patients. Non pharmacological interventions can reduce blood pressure and should be recommended to every patient suffering from hypertension. Nevertheless, a drug therapy must often be introduced. The threshold at which a treatment should be started depends mostly on the cardiovascular risk of each patient and the benefit of antihypertensive drug therapy depends primarily on the reduction of the blood pressure itself. PMID- 15898678 TI - [Multiple diseases: should we treat them all?]. AB - Studies are demonstrating additional benefits of multiple drug treatment in specific diseases. However, when a patient suffers multiple diseases, the lack of adequate clinical data on which to base a therapeutic attitude is baffling. The practitioner is forced to prescribe without a strong evidence base or to withhold medications for fear of doing more harm than good. In such circumstances, a prudent and drug sparing approach is to be preferred: a patient, not disease, oriented approach, using a few principles of rational prescribing: clear therapeutic objectives, prioritisation according to the severity of diseases, efficacy and safety of available therapies, therapeutic individualisation and monitoring, patient implication and attention to his/her desires and expectations. PMID- 15898679 TI - [The selection of a drug in a defined therapeutic class: the case of the HMG-CoA reductase inhibitors]. AB - The HMG-CoA reductase inhibitors have similar therapeutic targets and indications. However, their potential pharmacokinetic drug-drug interaction profile may play a significant role in their safety profile in polymedicated and polymorbid patients and can serve as a selection criterion. If their utility is clearly demonstrated in selected conditions, their safety profile remains of concern. Beside dose-related hepatic and muscular injury, other rare and important adverse drug reactions have been reported after prolonged administration such as polyneuropathy, fibrotic interstitial pulmonary disease and lupus-like syndrome. Teratogenicity has also been associated with statin therapy. PMID- 15898680 TI - [Some preconceptions and psychotropic drugs prescription]. AB - Prescription of psychotropic drugs should follow evidence-based medicine, i.e. be founded on well-conducted randomised controlled trials. In fact, a number of prescriptions do not follow these rules and cannot be qualified as evidence-based psychiatry. In this article, some preconceptions concerning the drug prescription in psychiatry are evoked. PMID- 15898681 TI - [The blood-brain barrier: recent insights]. AB - The blood-brain barrier must not be regarded as a purely passive wall opposing low permeability to undesirable compounds circulating in blood. It limits the access of molecules to brain tissue by a joint interplay of cerebro-spinal fluid turnover and selective uptake and expulsion of compounds through active drug carriers. The inhibition of such transporters can increase brain penetration of substrates leading to drug interactions with neuropsychiatric manifestations; it can also be exploited to increase brain and CSF concentrations of anti-infective, anti-neoplastic or psychotropic agents. Genetic polymorphisms affecting transporters may modulate the brain exposure to drugs in selected individuals. Such observations improve our understanding of a key mechanism governing clinical neuro-psychopharmacology. PMID- 15898682 TI - [The PROVE IT-TIMI-22 study or the biter being bit]. AB - The importance of statins in secondary prevention is well established since the 4S, CARE and LIPID studies, their indications being now applied to even normocholesterolemic patients. To date, it is still unclear which statin to choose, and at what dose. A recent study entitled "PROVE IT-TIMI-22" has compared 80 mg of atorvastatin/day to 40 mg of pravastatin/day in early secondary prevention. It appears that the intensive treatment with atorvastatin has been more effective on the LDL-cholesterol levels and has had a more favourable effect on the clinical evolution based on a composite score. We herein propose a critical review of this study and recommend a somewhat cautious attitude before giving high doses of atorvastatin in the secondary prevention of all the patients with coronary heart disease. PMID- 15898683 TI - [Willow bark extract: more than a natural alternative for the treatment of rheumatism?]. PMID- 15898685 TI - [Numbers and beings]. PMID- 15898686 TI - [Facing the announced death... choice between quality of life or headlong rush]. PMID- 15898687 TI - [Colon cancer: a new French medical affair?]. PMID- 15898688 TI - ["Mental distress," brief French tragicomedy (Act I)]. PMID- 15898689 TI - [Disruptive technologies and the Pope]. PMID- 15898690 TI - Beyond the water towers. PMID- 15898691 TI - Whole in one. PMID- 15898692 TI - Living with dementia. PMID- 15898693 TI - We are here. PMID- 15898694 TI - It helped to build me up, knowing that she been there and had got through it herself. PMID- 15898695 TI - It's a crime when people are written off but these reforms will make larger holes in the safety net. PMID- 15898696 TI - Institutionalised racism lies at the heart of the conceptual systems we use in psychiatry. PMID- 15898697 TI - What's funny about phobias? PMID- 15898698 TI - Running for your life. PMID- 15898699 TI - Workforce or workfare? PMID- 15898700 TI - What makes people crazy. PMID- 15898701 TI - This life. PMID- 15898702 TI - Formulation and in vitro/in vivo evaluation of combining DNA repair and immune enhancing nutritional supplements. AB - Combining nutritional supplements to achieve synergistic benefit is a common practice in the nutraceutical industry. However, establishing added health benefit from a combination of natural ingredients is often assumed, untested and without regard to the principle of metabolic competition between the active components. Here, we report on the combination of a cat's claw water extract (C Med-100, carboxy alkyl esters = active ingredients) + medicinal mushroom extracts (Cordyceps sinensis, Grifola blazei, Grifolafrondosa, Trametes versicolor and Ganoderma lucidum, polysaccharides = active ingredients) + nicotinamide + zinc into a formulation designed to optimize different modes of immunostimulatory action, and yet that would avoid metabolic antioxidant competition yielding less than expected efficacious effects. Isobole curve analyses of these two active classes of ingredients determined by growth inhibition of HL-60 human leukemic cells in vitro confirmed they were indeed synergistic when in combination, and not metabolically competitive. Furthermore, an in vivo study showed significant health benefit for 14 subjects treated for 4 weeks with the unique C-Med 100/mushroom extract formulation in that they had reduced pain, reduced fatigue, weight loss and a reduced presence of DNA damage in peripheral blood assessed by (8-OH) guanine DNA adducts and elevation in serum protein thiols. Because this broad-based panel of clinical parameters indicating clinical efficacy has never been demonstrated before for either of the active ingredients evaluated alone in humans, these data were taken as strong evidence that the combination of C-Med 100 + mushroom extracts + nicotinamide + zinc gave additive or synergistic effects to health benefit, and thus supported no efficacious limits from metabolic competition regarding this particular formulation. PMID- 15898703 TI - Anti-ulcer and anti-oxidant activity of pepticare, a herbomineral formulation. AB - Pepticare, a herbomineral formulation of the Ayurveda medicine consisting of the herbal drugs: Glycyrrhiza glabra, Emblica officinalis and Tinospora cordifolia, was tested for its anti-ulcer and anti-oxidant activity in rats. Effects of various doses (125, 250, 500 and 1000mg/kg, p.o.) of Pepticare were studied on gastric secretion and gastric ulcers in pylorus-ligation and on ethanol-induced gastric mucosal injury in rats. The reduction in ulcer index in both the models along with the reduction in volume and total acidity, and an increase in the pH of gastric fluid in pylorus-ligated rats proved the anti-ulcer activity of Pepticare. It was also found that Pepticare was more potent than G. glabra alone in protecting against pylorus-ligation and ethanol-induced ulcers. The increase in the levels of superoxide dismutase, catalase, reduced glutathione and membrane bound enzymes like Ca2+ ATPase, Mg2+ ATPase and Na+ K+ ATPase and decrease in lipid peroxidation in both the models proved the anti-oxidant activity of the formulation. Thus it can be concluded that Pepticare possesses anti-ulcer activity, which can be attributed to its anti-oxidant mechanism of action. PMID- 15898704 TI - Extracts and constituents of Lavandula multifida with topical anti-inflammatory activity. AB - The topical anti-inflammatory activity of the ethanol and aqueous extracts from the aerial parts of Lavandula multifida L. (Lamiaceae), used in the Moroccan traditional medicine, was investigated by inhibition of the Croton oil-induced ear edema in mice. The biological assay revealed a dose-dependent anti inflammatory activity for the ethanol extract, while the aqueous one was less active. Bioassay-guided fractionation of the ethanol extract led to identify four triterpenic acids of oleanane series, four pimarane and one iso-pimarane diterpenes, as well as the phenolic monoterpene carvacrol and its glucoside. Some of these compounds revealed an anti-inflammatory activity comparable to that of indomethacin. PMID- 15898705 TI - Activity of taraxasteryl acetate on inflammation and heat shock protein synthesis. AB - Pluchea sagittalis whole plant dichloromethane extract showed inhibitory activity in several inflammatory models: rat hind paw-edema, mice ear edema, and air-pouch rat granuloma. The extract inhibited the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in stimulated human neutrophils. It also showed inhibitory effect on heat shock protein 72 (hsp72) synthesis in stimulated neutrophils, while it had opposite effects on unstimulated cells. The triterpene taraxasteryl acetate was obtained from the dichloromethane extract by bioassay directed isolation, being active against induced ROS and RNS production in human neutrophils. In mice ear edema (induced by phorbol-12-mirystate-13 acetate, croton oil and arachidonic acid), taraxasteryl acetate showed a topical anti-inflammatory activity similar to the extract, but at 1/20 of the dose. The same ratio was observed for the inhibition of hsp72 production in stimulated human neutrophils. In unstimulated monocytes and neutrophils, taraxasteryl acetate showed a higher stimulating activity of hsp72 production than the extract, involving different mechanisms in each cell type. To our knowledge, taraxasteryl acetate is the first natural product for which a dual effect on the hsp response is reported. PMID- 15898706 TI - Modulatory influence of Adhatoda vasica Nees leaf extract against gamma irradiation in Swiss albino mice. AB - The radiomodulatory influence of ethanolic extract of Adhatoda vasica Nees leaf extract against radiation-induced hematological alterations in peripheral blood of Swiss albino mice was studied at various post-irradiation intervals between 6 h to 30 days. Oral administration of A. vasica leaf extract (800 mg/kg body weight) prior to whole body irradiation showed a significant protection in terms of survival percentage and hematological parameters. Mice exposed to radiation (8.0 Gy) without A. vasica leaf extract pre-treatment exhibited signs of radiation sickness like anorexia, lethargicity, ruffled hairs and diarrhoea and such animals died within 25 days post-irradiation. The dose reduction factor (DRF = 1.6) for A. vasica leaf extract was calculated from LD50/30 values. A significant decline in hematological constituents (RBCs, WBCs, Hb and Hct) was evident till day 15 and no animal could survive beyond day 25. Conversely, animals pre-treated with A. vasica leaf extract showed 81.25% survival till 30 days after exposure and a gradual recovery was noted in the hematological values. However, these hematological values remained significantly below the normal even till day 30. A significant decrease in blood reduced glutathione (GSH) content and increase in lipid peroxidation (LPO) level was observed in control animals (Radiation alone). However, A. vasica leaf extract pretreated irradiated animals exhibited a significant increase in GSH content and decrease in LPO level. A significant increase in the serum alkaline phosphatase activity and decrease in acid phosphatase activity was observed in A. vasica leaf extract pretreated irradiated animals during the entire period of study. PMID- 15898707 TI - Effects of methyl-eugenol administration on behavioral models related to depression and anxiety, in rats. AB - Croton zehntneri (Cz) is a popular plant in Brazilian folk medicine. Recently, the use of its essential oil showed depressive activity response in the central nervous system (CNS). Chemical studies show that the main compound of this oil is the methyl-eugenol (ME). This work seeks to evaluate the ME activity in behavioral models of depression and anxiety, in the rat. Male rats (60 days old) were divided into four groups (n = 10) and treated with doses of 1.0, 3.0 and 10.0 ml/100 g body wt., v.o., of ME (experimental) and saline (control). One hour after treatment, they were observed in the forced swimming test and 15 min later in the open-field test. A decrease was observed in the immobility time during the forced swimming test for all experimental groups, in comparison with control group (C = 168.8 +/- 27.3; 1.0 microl = 139.1 +/- 23.5; 3.0 microl = 137.2 +/- 18.7 and 10.0 microl = 139.8 +/- 23.6). The open-field results showed no differences in comparison to the control group. The same was observed for social interaction, plus-maze and holeboard tests, suggesting no alterations in anxiety behavior. These data suggest that ME administration induced antidepressive CNS alterations, expressed by the smallest immobility in the swimming model, and not of a level able to alter motor and exploratory activity in the open-field. The absence of effects observed in the open-field can be a result of the experimental contingency, taking low anxiety levels. These data are in contradiction to observations with Cz essential oil in these models. PMID- 15898708 TI - Chemical composition and anticancer activity of leaf essential oil of Myrica gale L. AB - Myrica gale L. (Myricaceae), a native plant from Canada used in traditional medicine, was extracted by hydrodistillation and the oil was collected after 30 and 60 min. The chemical composition of these two extracts was determined using GC-MS analysis. We identified 53 components and myrcene (23.18-12.14%), limonene (11.20-6.75%), alpha-phellandrene (9.90-6.49%) and beta-caryophyllene (9.31 10.97%) were the major components in the 30- and 60-min fractions, respectively, whereas higher caryophyllene oxide content was detected in the 60-min fraction (9.94%) than in the 30-min fraction (3.47%). The anticancer activities of these extracts were assessed against human lung carcinoma cell line A-549 and human colon adenocarcinoma cell line, DLD-1. The 60-min fraction showed higher anticancer activity against both tumor cell lines with an IC50 value of 88 +/- 1 microg/ml. The 30-min fraction had an IC50 value of 184 +/- 4 microg/ml for A-549 and 160 +/- 3 microg/ml for DLD-1. The higher cell growth inhibition induced by the 60-min fraction, as compared to the 30-min fraction, could be due to sesquiterpene enrichment. PMID- 15898709 TI - Bacopa monniera, a reputed nootropic plant: an overview. AB - Bacopa monniera (BM), a traditional Ayurvedic medicine, used for centuries as a memory enhancing, anti-inflammatory, analgesic, antipyretic, sedative and antiepileptic agent. The plant, plant extract and isolated bacosides (the major active principles) have been extensively investigated in several laboratories for their neuropharmacological effects and a number of reports are available confirming their nootropic action. In addition, researchers have evaluated the anti-inflammatory, cardiotonic and other pharmacological effects of BM preparations/extracts. Therefore, in view of the important activities performed by this plant, investigation must be continued in the recently observed actions described in this paper. Moreover, other clinical studies have to be encouraged, also to evidence any side effects and possible interactions between this herbal medicine and synthetic drugs. PMID- 15898710 TI - Biological activities of Ginkgo extracts. AB - The biological activity of methanolic the extracts of leaves, roots, leaf-derived callus, root-derived callus, ginkolide A, ginkgolide B, bilobalide and a commercial Ginkgo product (Tanakan) was assessed. Bioassays consisted of the Agrobacterium tumefaciens-induced potato tumor assay and a Kirby-Bauer microbial sensitivity assay with pure strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pyogenes. Methanolic extracts of leaves, leaf-derived callus, root derived callus, bilobalide and Tanakan inhibited tumor formation significantly, but more weakly than the positive control, camptothecin. No activity against E. coli was detected, but extracts from both callus types inhibited the growth of K. pneumonia, P. aeruginosa, S. aureus, S. epidermidis and S. pyogenes. All extracts and reference compounds inhibited the growth of S. pyogenes. Leaf and root tissues contained the highest levels of ginkgolide A, as compared to the callus tissues; leaf tissue contained more of all three marker compounds than the callus tissues. PMID- 15898711 TI - Use of carbonyl group addition--elimination reactions for synthesis of nucleic acid conjugates. AB - This review outlines the synthesis of covalent conjugates of oligonucleotides and their analogues that are obtained by reactions of carbonyl compounds with various nucleophiles such as primary amines, N-alkoxyamines, hydrazines, and hydrazides. The products linked by imino, oxime, hydrazone, or thiazolidine groups are shown to be useful intermediates for a wide range of chemical biology applications. Methods for their preparation, isolation, purification, and analysis are highlighted, and the comparative stabilities of the respective linkages are evaluated. The relative merits of incorporation of a carbonyl group, particularly an aldehyde group, into either the oligonucleotide or the ligand parts are considered. Examples of harnessing of aldehyde-nucleophile coupling for the labeling of nucleic acids are given, as well as their conjugation to various biomolecules (e.g. peptides and small molecule ligands), site-specific cross linking of oligonucleotides to nucleic acid-binding proteins, assembly of multibranched supramolecular structures, and immobilization on functionalized surfaces. Future perspectives of bioconjugation and complex molecular engineering via carbonyl group addition-elimination reactions in nucleic acids chemistry are discussed. PMID- 15898712 TI - Inhibition of membrane-associated methyltransferases by a cholesterol-based metal chelator. AB - We have designed, synthesized, and characterized a metal chelating compound that is based on the structure of cholesterol and contains the high affinity metal chelating group, lysine nitrilotriacetic acid (Lys-NTA). Using the enzyme isoprenylcysteine carboxylmethyltransferase (Icmt) from yeast as a model integral membrane metalloenzyme, we find that this agent potently inhibits Icmt activity with an IC(50) value between 35 and 75 microM, which is at least 40 times more potent than the best known Icmt metal chelating inhibitor, Zincon. We propose that the rigid hydrophobic cholesterol moiety promotes partitioning into the membrane, enabling the metal-binding NTA group(s) to inactivate the enzyme by metal chelation. Because this compound is based on a naturally occurring membrane lipid and appears to chelate metals buried deeply within water insoluble environments, this agent may also be useful as a general tool for identifying previously unappreciated metal dependencies of other classes of membrane proteins. PMID- 15898713 TI - Transferrin-mediated gold nanoparticle cellular uptake. AB - Targeted drug delivery is an important research area in specific therapy. Transferrin-conjugated nanoparticles are an attractive formulation as a vehicle for specific cellular uptake and targeted drug delivery. In this report, atomic force microscopy imaging was used to visualize the process of cellular uptake of transferrin-coupled gold nanoparticles on the surfaces of live cells for the first time. High-resolution images were captured, showing the endocytosis of transferrin-conjugated nanoparticles taking place during the process of internalization. This specific transferrin-mediated nanoparticle uptake was validated by confocal scanning imaging and transferrin competition experiments. PMID- 15898714 TI - The peptide route to multifunctional gold nanoparticles. AB - Extremely stable, peptide-capped gold nanoparticles with two different biomolecular recognition motifs expressed on their surface have been prepared, and their specific and selective binding to artificial, DNA-modified target particles and to DNA and protein microarrays has been demonstrated. Stabilization and biofunctionalization has been achieved in a single preparative step starting with citrate-stabilized gold hydrosols and a derivatization cocktail of peptide capping ligands, which carry the functionalities of choice. PMID- 15898715 TI - Enhanced oligonucleotide binding to self-assembled nanofibers. AB - A peptide nucleic acid/peptide amphiphile conjugate (PNA-PA) that self-assembles into fiber-shaped nanostructures was designed to bind oligonucleotides with high affinity and specificity. Oligonucleotide binding to PNA-PA nanofibers was studied using fluorescence polarization, and thermal stability was examined by UV vis measurement of duplex melting temperatures. The self-assembled PNA-PA DNA system was observed to bind more strongly than the corresponding DNA-DNA duplex. We also observed single base specificity with a 16 degrees C in thermal stability. As expected from the previous PNA studies, PNA-PA RNA binding is also stronger than the corresponding RNA-RNA duplex. PMID- 15898716 TI - Structural and biophysical characterization of the 40 kDa PEG-interferon-alpha2a and its individual positional isomers. AB - The human recombinant Interferon-alpha(2a) (IFNalpha(2a)) is a potent drug (Roferon-A) to treat various types of cancer and viral diseases including Hepatitis B/C infections. To improve the pharmacological properties of the drug, a new pegylated form of IFNalpha(2a) was developed (PEGASYS). This 40 kDa PEG conjugated IFNalpha(2a) ((40)PEG-IFNalpha(2a)) is obtained by the covalent binding of one 40 kDa branched PEG-polymer to a lysine side chain of IFNalpha(2a). (40)PEG-IFNalpha(2a) is a mixture of mainly six monopegylated positional isomers modified at K31, K134, K131, K121, K164, and K70, respectively. Here we report the detailed structural and biophysical characterization of (40)PEG-IFNalpha(2a) and its positional isomers, in comparison with IFNalpha(2a), using NMR spectroscopy, analytical ultracentrifugation, circular dichroism, fluorescence spectroscopy, and differential scanning calorimetry. Our results show that the three-dimensional structure of IFNalpha(2a) is not modified by the presence of the polymer in all positional isomers constituting (40)PEG-IFNalpha(2a). Regardless of where the PEG polymer is attached, it adopts a very mobile and flexible random coil conformation, producing a shield for the protein without a permanent coverage of the protein surface. Hydrodynamic data indicate that the protein-attached PEG has a slightly more compact random-coil structure than the free PEG-polymer. Our results also provide evidence of significant structural and physicochemical advantages conferred by the pegylation: increase of the effective hydrodynamic volume and modification of the molecular shape, higher temperature stability, and reduced tendency for aggregation. These results are of tremendous pharmacological interest and benefit as was clinically shown with PEGASYS. This study constitutes a new standard for the characterization of pegylated proteins and enables an important step toward the understanding on a molecular level of the binding of (40)PEG-IFNalpha(2a) and its positional isomers to its cellular receptors. PMID- 15898717 TI - Structural, kinetic, and thermodynamic analysis of the binding of the 40 kDa PEG interferon-alpha2a and its individual positional isomers to the extracellular domain of the receptor IFNAR2. AB - Type-I Interferons exert antiviral and antiproliferative activities through the binding to a common cell surface receptor comprising two subunits, IFNAR1 and IFNAR2. Human recombinant Interferon-alpha(2a) (IFNalpha(2a)) is a potent drug (Roferon-A) used to treat various cancers and viral diseases including Hepatitis B/C infections. To significantly improve the pharmacological properties of the drug, a pegylated form of IFNalpha(2a) was developed (PEGASYS). This 40 kDa PEG conjugated IFNalpha(2a) ((40)PEG-IFNalpha(2a)) is obtained by the covalent binding of one 40 kDa branched PEG-polymer to a lysine side-chain of IFNalpha(2a). Here, we report the detailed structural, kinetic, and thermodynamic analysis of the binding to the extracellular domain of the receptor IFNAR2 of (40)PEG-IFNalpha(2a) and its isolated positional isomers modified at K31, K134, K131, K121, K164, and K70, respectively, in comparison with unmodified IFNalpha(2a). Our binding studies, using the surface plasmon resonance technique, show that the pegylation does not abolish the binding to the receptor, but significantly reduces the affinity mainly due to a change of the association rate. The results are supported by modeling and simulation of the binding, using Self-Avoiding-Walk calculations for the polymer conformations. A correlation between the structural parameters and the kinetic and thermodynamic parameters of the binding of the positional isomers could be established. For the Isomer-K31 and -K164, the PEG-polymer attachment point is located in proximity to the binding interface, and the isomers display affinity in the range 150-520 nM in an enthalpy-driven binding process. In contrast for the Isomer-K134, -K131, -K121, and -K70, the PEG-polymer is attached remotely from the binding interface, and the isomers exhibit a higher affinity (32-76 nM) in an entropy-driven binding process. This study constitutes an essential collection of knowledge on which the interaction of (40)PEG-IFNalpha(2a) and its positional isomers with its cellular receptors can be better understood. PMID- 15898718 TI - Oligomeric fluorescent labels for DNA. AB - In an effort to find fluorescent labels that have large Stokes shifts and increased emission intensity, a strategy for fluorescence labeling of DNA was explored in which multiple individual fluorophores are incorporated at adjacent positions at the end of a DNA probe. To encourage close interactions, hydrocarbon and heterocycle fluorophores were substituted at C-1 of deoxyribose, replacing the DNA base. The C-glycosides studied contained the well-known fluorophores terphenyl, pyrene, and terthiophene. For comparison, a commercial fluorescein-dU nucleotide was examined. Oligomeric labels containing up to five fluorophores were tested. Interestingly, all four dyes behaved differently on multiple substitution. Fluorescein displayed strong self-quenching properties, with the quantum yield dropping severalfold with each additional substitution and with a constant, small Stokes shift. In contrast, pyrene showed increases in quantum yield on addition of more than one fluorophore and yielded efficient long wavelength emission on multiple substitution, with Stokes shifts of >130 nm. Oligomeric terphenyl labels gave a small progressive red shift in absorption and a marked red shift in emission wavelength and showed a strong increase in brightness with more monomers. Finally, terthiophene oligomers showed self quenching combined with increasing Stokes shifts. Overall, the results suggest that some oligomeric fluorescent labels exhibit properties not available in common fluorescein class (or other commercial) labels, such as large Stokes shifts and increasing brightness with increasing substitution. PMID- 15898719 TI - In vitro and in vivo evaluation of hydrophilic dendronized linear polymers. AB - Rigid-rod dendronized linear polymers consisting of a poly(4-hydroxystyrene) backbone and fourth-generation polyester dendrons were evaluated in vitro and in vivo to determine their suitability as drug delivery vectors. Cytotoxicity assays indicated that the polymers were well tolerated by cells in vitro. Biodistribution studies of the polymers in both nontumored and tumored mice revealed that as for random coil linear polymers, renal clearance was a function of polymer size, with significant urinary excretion observed for a 67 kDa dendronized polymer. High accumulation in organs of the reticuloendothelial system was exhibited by a dendronized polymer with a very high molecular weight (M(n) = 1740 kDa), but was not as significant for smaller polymers with M(n) = 67 kDa and M(n) = 251 kDa. The rank order for tumor accumulation of the polymers on a percent injected dose per gram tumor basis was 251 kDa approximately 1740 kDa > 67 kDa. These data will help guide the selection of highly functionalizable rigid rod dendronized polymers with pharmacokinetic properties appropriate for use as drug carriers. PMID- 15898720 TI - Near-infrared optical imaging of B16 melanoma cells via low-density lipoprotein mediated uptake and delivery of high emission dipole strength tris[(porphinato)zinc(II)] fluorophores. AB - Meso-to-meso ethyne-bridged tris[(porphinato)zinc(II)] (PZn(3)) near-infrared (NIR) fluorophores (lambda(em)(max) approximately 800 nm) can be rendered sufficiently amphiphilic to enable their facile incorporation into the hydrophobic core of the apo form of low-density lipoprotein (apo-LDL). These NIR fluorophores are notable in that they manifest low energy excited states polarized exclusively along the long axis of the supermolecule, broad spectral coverage of the visible and high energy NIR spectral domains, intense S(0)-->S(1) transition moments, and comparably large S(1)-->S(0) emission dipole strengths. The reconstituted LDL(PZn(3)) proteins can be used to deliver rapidly hundreds of copies of PZn(3) to a given murine B16 melanoma cell via LDL receptor-mediated endocytosis. PZn(3)-based NIRFs and their corresponding LDL(PZn(3)) proteins have been shown to display minimal cytotoxicity. Confocal NIR fluorescence microscopy evinces that B16 cells can be imaged at very low doses (approximately nM) of NIRF. The highly attractive photophysical properties of PZn(3) and closely related chromophores, coupled with their lack of toxicity and compatibility with uptake into apo-LDL and subsequent rapid delivery to B16 cells via LDLr-mediated endocytosis, suggest the potential utility of this platform for NIR optical imaging of cancer cells in vivo. PMID- 15898721 TI - Development of polymer latex particles for selective cleavage of mismatched DNA and their application for DNA diagnosis. AB - We developed functional polymer latex particles that can catch and cleave mismatched DNA selectively and propose a new mismatch detection system using the functional particles. The aimed particles possess two functional units composed of mismatch binding protein (MutS) and an anthraquinone derivative (AQ), a light activated agent that photocleaves dsDNA. Use of the functional particles made it possible to discriminate complementary and mismatched DNAs and photocleave mismatched DNA selectively. The efficiency of photocleavage of mismatched DNA by the functional particles increased with UV irradiation time. It was also found that the functional particles were reusable and had dissociation constants (K(d)) of 1000 and 68.5 nM for G/C homoduplex and G/T heteroduplex, respectively. Using the functional particles and a dsDNA-binding fluorescent dye, SYBR-Gold, we could construct the system for detection of mismatched DNA that was 40 base pairs. The functional particles prepared in this study will be an absolutely new tool for mismatch detection in DNA diagnosis. PMID- 15898722 TI - Pegylated, steptavidin-conjugated quantum dots are effective detection elements for reverse-phase protein microarrays. AB - Protein microarray technologies provide a means of investigating the proteomic content of clinical biopsy specimens in order to determine the relative activity of key nodes within cellular signaling pathways. A particular kind of protein microarray, the reverse-phase microarray, is being evaluated in clinical trials because of its potential to utilize limited amounts of cellular material obtained through biopsy. Using this approach, cellular lysates are arrayed in dilution curves on nitrocellulose substrates for subsequent probing with antibodies. To improve the sensitivity and utility of reverse-phase microarrays, we tested whether a new reporter technology as well as a new detection instrument could enhance microarray performance. We describe the use of an inorganic fluorescent nanoparticle conjugated to streptavidin, Qdot 655 Sav, in a reverse-phase protein microarray format for signal pathway profiling. Moreover, a pegylated form of this bioconjugate, Qdot 655 Sav, is found to have superior detection characteristics in assays performed on cellular protein extracts over the nonpegylated form of the bioconjugate. Hyperspectral imaging of the quantum dot microarray enabled unamplified detection of signaling proteins within defined cellular lysates, which indicates that this approach may be amenable to multiplexed, high-throughput reverse-phase protein microarrays in which numerous analytes are measured in parallel within a single spot. PMID- 15898723 TI - Selective cell attachment to a biomimetic polymer surface through the recognition of cell-surface tags. AB - Reactive phosphorylcholine polymers, which can recognize biosynthetic cell surface tags, were synthesized to control cell attachment. Human promyelocytic leukemia cells (HL-60) with unnatural carbohydrates as cell-surface tags were harvested by treatment with N-levulinoylmannosamine (ManLev). The attachment of ManLev-treated HL-60 cells to 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers with hydrazide groups was studied. HL-60 cells, which are nonadhesive, did not attach to any polymer surface without ManLev treatment. In contrast, ManLev-treated HL-60 cells attached to a poly[MPC-co-n-butyl methacrylate (BMA) co-methacryloyl hydrazide (MH)] (PMBH) surface following 15 min of incubation. The cells that attached to the PMBH surface retained their native morphology and viability for 24 h of incubation. On the other hand, approximately half of the HL 60 cells that attached to the poly(BMA-co-MH) (PBH) surface died. These results suggest that MH units in the polymer act as anchors for cell attachment and MPC units help to preserve cell viability on a polymer surface. The coculture of ManLev-treated HL-60 and fluorescence-stained human uterine cervical cancer cells (HeLa) was carried out on polymer surfaces. ManLev-treated HL-60 cells specifically attached to the PMBH surface. In contrast, both HL-60 and HeLa cells were observed on the PBH surface. The control of cellular interactions with synthetic polymers may be useful for the future development of cell-integrated biosensors and biomedical devices. PMID- 15898724 TI - In vivo imaging of activated endothelium using an anti-VCAM-1 magnetooptical probe. AB - It has been suggested that vascular cell adhesion molecule-1 (VCAM-1) could serve as an early marker for inflammation of the endothelium. The ability to noninvasively image VCAM-1 could thus be a useful tool to diagnose a number of inflammatory diseases at early stages. Here we demonstrate that magnetooptical nanoparticles conjugated to anti-VCAM-1 antibodies can be used to specifically detect VCAM-1 expression on endothelial cells in culture and in vivo. Elevated VCAM-1 expression was detected on cultured murine heart endothelial cells by both fluorescence and magnetic resonance, while only basal expression levels were detected on murine dermal endothelial cells. Intravital microscopy of a murine inflammatory model injected with the VCAM-1 targeted nanoparticles revealed specific labeling of the activated endothelium, with labeling kinetics yielding a maximum vessel wall signal 6 h after injection. In contrast, nontargeted nanoparticles did not exhibit any specific labeling of the endothelium. These studies suggest that the developed nanoparticle would be useful for MR and optical detection of activated endothelium. PMID- 15898725 TI - Cell uptake and radiotoxicity studies of an nuclear localization signal peptide intercalator conjugate labeled with [99mTc(CO)3]+. AB - A trifunctional bioconjugate consisting of the SV40 nuclear localization signal (NLS) peptide, an aliphatic triamine ligand, and the DNA intercalating pyrene has been synthesized and quantitatively labeled with [(99m)Tc(OH(2))(3)(CO)(3)](+). The radiotoxicity of the resulting nucleus-targeting radiopharmaceutical on B16F1 mouse melanoma cells has been investigated to evaluate the activity of Auger and Coster-Kronig electrons on the viability of cells. We found a dose-dependent significant radiotoxicity of the nucleus-targeting radiopharmaceutical clearly related to the low energy decay of (99m)Tc. These principal results imply a possible therapeutic strategy based on the use of the low-energy Auger electron emitting (99m)Tc radionuclide attached to nucleus-targeting molecules and comprising an intercalator. Highly efficient DNA targeting vectors could complement the usual role of (99m)Tc in diagnostic applications. The Auger electrons emitted by the (99m)Tc nuclide induce DNA damage leading ultimately, through a mitotic catastrophe pathway, to necrotic cell death. Non-DNA-targeting (99m)Tc complexes display much lower radiotoxicity. PMID- 15898726 TI - Cross-linking between thymine and indolyl radical: possible mechanisms for cross linking of DNA and tryptophan-containing peptides. AB - As experimentally observed in gamma-irradiated aqueous solutions of tryptophan containing peptides in the presence of DNA, a fast electron (or hydrogen atom) transfer from the DNA restores an intact tryptophan residue at the expense of the DNA integrity. Alternatively, addition of the deprotonated electron-deficient indolyl radical to the DNA, followed by subsequent rearrangement, may lead toward DNA/tryptophan-containing peptide cross-linking. Herein, possible reaction mechanisms for thymine-indolyl radical cross-linking are proposed. The consistent use of the contact spin density distribution is the key virtue of this work. The Becke 3, Lee, Yang, and Parr (B3LYP) density functional theory (DFT) method is employed to investigate the feasibility of the proposed cross-linking mechanisms. A possible complete reaction mechanism consists of a combination of the C(5) hydroxylated thymine and indolyl radicals forming the initial cross-linked product, a hydrogen transfer within the initial cross-linked product by use of a bridging water molecule, and a dehydration step. The overall thermodynamics of the free energy profiles at 0 and 298 K are similar and display differences of magnitude for the hydrogen-transfer reaction. Temperature may be a key factor influencing the overall mechanism. The skeletal structures and contact spin densities on the heavy atoms of the tryptophan side chain and indolyl radicals are essentially equal. Hence, it is believed that a direct combination of the C(5)-hydroxylated thymine and tryptophan radicals should form the initial cross linked product, as far as addition of the tryptophan radical to the DNA is concerned. PMID- 15898727 TI - N-(2-hydroxypropyl)methacrylamide copolymers of a glutathione (GSH)-activated glyoxalase i inhibitor and DNA alkylating agent: synthesis, reaction kinetics with GSH, and in vitro antitumor activities. AB - The incorporation of anticancer prodrugs into polyacrylamide conjugates has been shown to improve tumor targeting via the so-called "enhanced permeability and retention" effect. This strategy has now been expanded to include two different classes of glutathione (GSH)-activated antitumor agents prepared by radical polymerization of N-(2-hydroxypropyl)methacrylamide (HPMA) with 2-methacryloyloxy methyl-2-cyclohexenone (7) and/or with S-(N-4-chlorophenyl-N-hydroxycarbamoyl thioethyl)methacrylamide (8), followed by treatment with 3-chloroperoxybenzoic acid, to give the HPMA copolymers of 7 and the 8-sulfoxide, respectively. In aqueous-buffered solution at pH 6.5, GSH reacts rapidly with poly-HPMA-8 sulfoxide (k approximately 2.3 mM(-1) min(-1)) to give S-(N-4-chlorophenyl-N hydroxycarbamoyl)glutathione (1), a tight-binding transition state analogue inhibitor of the antitumor target enzyme glyoxalase I (K(i) = 46 nM), or with poly-HPMA-7 (k approximately 0.02 mM(-1) min(-1)) to give the electrophilic antitumor agent 3-glutathio-2-methylenecyclohexenone (4). Indeed, B16 melanotic melanoma in culture is inhibited by poly-HPMA-8-sulfoxide and by poly-HPMA-7 with IC(50) values of 168 +/- 8 and 284 +/- 5 microM, respectively. These values are significantly greater than those of the unpolymerized prodrugs suggesting that the cytotoxicity of the polymer prodrugs might be limited by slow cellular uptake via pinocytosis. This prodrug strategy should be applicable to a range of different GSH-based antitumor agents. PMID- 15898728 TI - Neutral postgrafted colloidal particles for gene delivery. AB - Surface modification of cationic lipoplexes has been carried out by means of a postgrafting reaction. The original lipoplexes described comprise a cationic lipid, a neutral lipid, poly(ethylene glycol)-cholesterol (with or without a targeting ligand) and DNA. Modifying their surface via a chemical, postgrafting reaction did not alter their size (approximately 100 nm) nor their ability to compact DNA, but did give a reduced zeta potential (approximately 0 mV) to afford surface neutral particles. With the modified lipoplexes nonspecific NIH3T3 cell surface binding in vitro was inhibited. Intravenous injection of the neutralized lipoplexes in mice showed decreased accumulation of the particles in the lung as compared to PEGylated cationic lipoplexes. Tumor targeting was also achieved in vivo by the addition of an RGD-PEG-Cholesterol as a lipid-ligand in the postgrafted lipoplex formulation. PMID- 15898729 TI - Synthesis and biological properties of insulin-deoxycholic acid chemical conjugates. AB - Bile acids have been considered very useful in the preparation of new pharmaceuticals, and more recently in the preparation of peptide and protein drugs because of their natural chemical and biological properties. In this study, we modified recombinant human insulin by covalently attaching deoxycholic acid (DOCA) derivatives in order to synthesize orally active insulin analogues. DOCA derivatives, namely succinimido deoxycholate and succinimido bisdeoxycholyl-L lysine were prepared and site specifically conjugated at Lys(B29) of insulin. The resultant insulin conjugates, [N(B29)-deoxycholyl] insulin (Ins-DOCA) and [N(B29) bisdeoxycholyl-L-lysil] insulin (Ins-bisDOCA), were studied for their chemical, structural, and biological properties. Their chemical properties were determined by HPLC, MALDI-TOF mass spectroscopy, and dynamic light scattering. Lipophilicity and self-aggregation behavior of insulin conjugates were enhanced with increasing number of labeled bile acid. The far-ultraviolet region of circular dichroism spectra showed no significant change of the tertiary structure of insulin in aqueous solution due to conjugation. Competitive insulin binding assay with HepG2 cells revealed that monosubstituted insulin conjugates still retained high binding affinity to the insulin receptor. When the insulin conjugates were intravenously administered (0.33 IU/kg) to streptozotocin (STZ)-induced diabetic rats, the conjugates showed sustained biological activity for a longer period with the similar lowest blood glucose level (glucose nadir), compared to native insulin. In further studies, the resulting new insulin conjugates will be investigated for their oral efficiency as a long-acting insulin formulation for the treatment of diabetic patients. PMID- 15898730 TI - Detection of lipopolysaccharide and lipid a employing a spermine-pyrene conjugate. AB - An amphiphilic fluorescent spermine-pyrene conjugate (Sp-Py) is shown to interact with lipopolysaccharide (LPS) or lipid A in aqueous solution. This complexation was studied by UV/visible absorption and steady-state fluorescence spectroscopy. In the presence of LPS or lipid A, Sp-Py displayed self-aggregation, resulting in the appearance of chromophore dimer absorption and fluorescence at the expense of probe monomer features. The equivalent weight of LPS or lipid A and the binding constants for Sp-Py complexation were estimated. The change of the ratio of monomer to dimer emission with concentration of LPS or lipid A indicated that Sp Py is a sensitive fluorescence probe for the endotoxins in aqueous media. PMID- 15898731 TI - Polycationic block copolymers of poly(ethylene oxide) and poly(propylene oxide) for cell transfection. AB - A facile, one-step synthesis of cationic block copolymers of poly(2-N (dimethylaminoethyl) methacrylate) (pDMAEMA) and copolymers of poly(propylene oxide) (PPO) and poly(ethylene oxide) (PEO) has been developed. The PEO-PPO-PEO pDMAEMA (L92-pDMAEMA) and PEO-pDMAEMA copolymers were obtained via free radical polymerization of DMAEMA initiated by polyether radicals generated by cerium(IV). Over 95% of the copolymer fraction was of molecular mass ranging from 6.9 to 7.1 kDa in size, indicating the prevalence of the polyether-monoradical initiation mechanism. The L92-pDMAEMA copolymers possess parent surfactant-like surface activity. In contrast, the PEO-pDMAEMA copolymers lack significant surface activity. Both copolymers can complex with DNA. Hydrodynamic radii of the complexes of the L92-pDMAEMA and PEO-pDMAEMA with plasmid DNA ranged in size from 60 to 400 nm, depending on the copolymer/DNA ratio. Addition of Pluronic P123 to the L92-pDMAEMA complexes with DNA masked charges and decreased the tendency of the complex to aggregate, even at stoichiometric polycation/DNA ratios. The transfection efficiency of the L92-pDMAEMA copolymer was by far greater than that of the PEO-pDMAEMA copolymer. An extra added Pluronic P123 further increased the transfecton efficacy of L92-pDMAEMA, but did not affect that of PEO-pDMAEMA. PMID- 15898732 TI - Mixed-ligand rhenium-188 complexes with tetradentate/monodentate NS3/P ('4 + 1') coordination: relation of structure with antioxidation stability. AB - Development of new radiopharmaceuticals based on rhenium-188 depends on finding appropriate ligands able to give complexes with high in vivo stability. Rhenium(III) mixed-ligand complexes with tetradentate/monodentate ('4 + 1') coordination of the general formula [Re(NS(3))(PRR'R' ')] (NS(3) = tris(2 mercaptoethyl)amine and derivatives thereof, PRR'R' ' = phosphorus(III) ligands) appear to be among the promising tools to achieve this goal. According to this approach, we synthesized and characterized a series of rhenium model complexes. In vitro stabilities of the corresponding rhenium-188 complexes were determined by incubating 2-3 MBq or alternatively 37 MBq of the complexes in phosphate buffer, human plasma, and rat plasma, respectively, at 22 degrees C or 37 degrees C, followed by checking the amount of (188)ReO(4)(-) formed after 1 h, 24, and 48 h by thin-layer chromatography. The rate of perrhenate formation varied over a wide range, depending primarily on the nature of the phosphorus(III) ligand. Physicochemical parameters of the corresponding nonradioactive rhenium complexes were analyzed in detail to find out the factors influencing their different stability and furthermore to design new substitution-inert '4 + 1' complexes. Tolman's cone angle of phosphorus(III) ligands and the lipophilic character of the inner coordination sphere were found to be crucial factors to build up stable rhenium '4 + 1' complexes. Additional information useful to describe electronic and steric properties of these compounds were selected from electronic spectra (wavelength of the Re-->S charge-transfer band), cyclovoltammetric measurements (E degrees of the Re(III)/Re(IV) couple), and NMR investigations ((31)P chemical shift of coordinated P(III) ligands). PMID- 15898733 TI - Radioiodinated azide and isothiocyanate derivatives of cocaine for irreversible labeling of dopamine transporters: synthesis and covalent binding studies. AB - Two novel N-substituted-3beta-phenyltropane alkaloids have been labeled with iodine-125 for use as irreversible probes of dopamine transporter (DAT) binding sites. One contains an iodoaryl azide moiety for photolabeling, while the other bears an iodoaryl isothiocyanate for direct conjugation. Both radioligands were prepared in a one-flask procedure by electrophilic radioiodination of the corresponding aniline under no-carrier-added conditions, followed either by diazotization and treatment with sodium azide, or by addition of thiophosgene under basic conditions. Specifically, (-)-N-[4-(3-[(125)I]iodo-4 azidophenyl)butyl]-2beta-carbomethoxy-3beta-(4-chlorophenyl)tropane ([(125)I]MFZ 2-24) and (-)-N-[4-(3-[(125)I]iodo-4-isothiocyanophenyl)butyl]-2beta-carbomethoxy 3beta-(4-chlorophenyl)tropane ([(125)I]MFZ 3-37) were synthesized. Isolation by reversed-phase HPLC and solid-phase extraction gave good average yields of [(125)I]MFZ-2-24 (67%, n = 5) and [(125)I]MFZ-3-37 (45%, n = 3) with high radiochemical purities (96-99%) and specific radioactivities (>2000 mCi/micromol). The utility of the radioligands was demonstrated by their covalent linkage to rat striatal membranes, and immunoprecipitation of a single radiolabeled band at 80 kDa corresponding to the full-length DAT. PMID- 15898734 TI - Adenosine 2'-monophosphate, 5'-O-[S-(4-succinimidylbenzophenone)-thiophosphate]: a new photoaffinity label for the coenzyme site of porcine NADP-specific isocitrate dehydrogenase. AB - A new photoaffinity label, adenosine 2'-monophosphate, 5'-O-[S-(4-succinimidyl benzophenone)thiophosphate] (2'-P-AMPS-Succ-BP), has been synthesized by an initial thiophosphorylation of 2'-AMP with PSCl(3) to form 2'-AMP-5' thiophosphate (2'-AMP-5'-SP), followed by a coupling reaction of 2'-AMP-5'-SP with benzophenone-4-maleimide to produce 2'-P-AMPS-Succ-BP. This product and its precursor were characterized by thin-layer chromatography, (31)P NMR, phosphorus analysis, and electron-spray mass spectroscopy. 2'-P-AMPS-Succ-BP functions as a photoaffinity label of porcine NADP-specific isocitrate dehydrogenase. To obtain reaction with other amino acids, Cys269 and Cys379, the most reactive cysteines of this enzyme, were mutated to yield a double mutant enzyme (C269A/C379S) exhibiting comparable activity and kinetic parameters to those of wild-type enzyme. 2'-P-AMPS-Succ-BP inactivates C269A/C379S enzyme upon UV irradiation. The reaction exhibits a nonlinear relationship of k(inact) versus [2'-P-AMPS-Succ-BP] with K(R) = 12 microM and k(max) = 0.0275 min(-1). NADP, NADPH, or 2'-monophospho adenosine 5'-diphosphoribose protects the enzyme against 2'-P-AMPS-Succ-BP inactivation. The ligand protection studies suggest that 2'-P-AMPS-Succ-BP binds to the porcine enzyme at the site best occupied by NADP/NADPH. The dimeric C269A/C379S isocitrate dehydrogenase incorporates 1.0 mol of 2'-P-[(35)S]AMPS Succ-BP/mol enzyme dimer concomitant with complete loss of enzyme activity. The new photoaffinity label may be generally useful to identify important amino acid residues of NADP-specific enzymes. PMID- 15898735 TI - Dramatic effect of the tridentate ligand on the stability of 99mTC "3 + 1" oxo complexes bearing arylpiperazine derivatives. AB - Mixed-ligand model complexes of general formula [(99m)Tc(O)(kappa(3) PNX)(kappa(1)-SPh))] [X = O (1a), S (2a)] were prepared in a one-step procedure from [(99m)TcO(4)(-)] using stannous chloride as reducing agent. Stability studies and challenge experiments with glutathione showed that complex 2a presented promising features for pursuing animal studies. The activity in the brain (% dose injected/organ) at 5 min (0.14% +/- 0.03) and 120 min (0.11% +/- 0.02) pi encouraged the synthesis of several mixed-ligand "3 + 1" oxo complexes of general formula [M(O)(kappa(3)-PNS)(kappa(1)-SL))] (M = (99m)Tc, 3a-6a, Re, 3 6), in which the tridentate ligand is the heterofunctionalized phosphine 2 (diphenylphosphanyl)-N-(2-thioethyl)benzamide (PNS) and the co-ligands are different arylpiperazine derivatives (HSL1-HSL4). The (99m)Tc complexes have been characterized by comparison of their retention times in the HPLC chromatogram (gamma-detection) with the retention times of the analogous Re complexes (UV detection at 254 nm). The (99m)Tc complexes, obtained with radiochemical purity higher than 95%, after HPLC purification, are stable in saline, 0.01 M PBS (pH 7.4), rat plasma (4 h, 37 degrees C), and glutathione (10 mM solutions, 2h, 37 degrees C). Binding affinity and selectivity for 5-HT(1A) receptors (relative to the 5-HT(2A) receptor) were determined, complex 5 demonstrating the best values (IC(50) for the 5-HT(1A) 2.35 +/- 0.02 nM; competitor 5-HT(2A) 372 +/- 11 nM). Biodistribution and stability studies in mice indicated a preferred hepatobiliary excretion, a high in vivo stability, but a poor brain uptake. PMID- 15898736 TI - Synthesis and activity of C11-modified wortmannin probes for PI3 kinase. AB - The key role played by PI3 kinase in cancer, hormone action, and a host of other biological functions suggests that specific inhibitors whose disposition could be ascertained in vivo would be useful in biological research or, potentially, for imaging PI3K in a clinical setting. Wortmannin (Wm, 1) is an inhibitor of PI3 kinase with high specificity for this enzyme. We synthesized three modified Wm probes, a biotinylated Wm (7a), a 4-hydroxy-3-iodophenylated Wm, which was obtained both unlabeled (7b) and labeled with (125)I (8), and a fluoresceinated Wm (7c), through modification at C-11, and evaluated their inhibitive activity as inhibitors of PI3 kinase. Biotinylated (7a) and 4-hydroxy-3-iodophenylated Wm's (7b) had IC(50)s for PI3K of 6.11 and 11.02 nM, respectively, compared to an IC(50) for Wm of 1.63 nM. Fluoresceinated Wm (7c) lost considerably more activity than the other derivatives, with an IC(50) of 64.9 nM. The (125)I labeled 4 hydroxy-3-iodophenylated Wm (8) could be detected after reaction with an immunoprecipitate of PI3 kinase. The activity of these reporter Wm's is discussed in relationship to earlier findings on the pharmacological activity of Wm derivatives and the ability of inhibitors to fit into the ATP pocket of PI3 kinase. PMID- 15898737 TI - Example of fatty acid-loaded lipoplex in enhancing in vitro gene transfer efficacies of cationic amphiphile. AB - Herein, we report on the design and synthesis of a novel nontoxic cationic amphiphile N,N-di-n-tetradecyl-N-[2-[N',N'-bis(2-hydroxyethyl)amino]ethyl]-N-(2 hydroxyethyl)ammonium chloride (lipid 1) whose in vitro gene transfer efficacies in CHO, COS-1, MCF-7, and HepG2 cells are remarkably enhanced when used in combination with 30 mole percent added myristic acid. Reporter gene expression assay using p-CMV-SPORT-beta-gal reporter gene revealed poor gene transfer properties of the cationic liposomes of lipid 1 and cholesterol (colipid). However, the in vitro gene delivery efficacies of lipid 1 were found to be remarkably enhanced when the cationic liposomes of lipid 1 and cholesterol were prepared in the presence of 30 mole percent added myristic acid (with respect to lipid 1) as the third liposomal ingredient. The whole cell histochemical X-gal staining of representative CHO cells further confirmed the significantly enhanced gene transfer properties of the fatty acid-loaded cationic liposomes of lipid 1 and cholesterol. Electrophoretic gel patterns in the gel mobility shift assay supports the notion that better DNA release from fatty acid lipoplexes might play a role in their enhanced gene transfer properties. In addition, such myristic acid-loaded lipoplexes of lipid 1 were also found to be serum-compatible up to 30% added serum. Taken together, our present findings demonstrate that the transfection efficacies of fatty acid-loaded lipoplexes are worth evaluating particularly when traditional cationic liposomes prepared with either cholesterol or DOPE colipids fail to transfect cultured cells. PMID- 15898738 TI - Sulfhydryl-reactive, cleavable, and radioiodinatable benzophenone photoprobes for study of protein-protein interaction. AB - The major task in proteomics is to understand how proteins interact with their partners. The photo-cross-linking technique enables direct probing of protein protein interaction. Here we report the development of three novel sulfhydryl reactive benzophenone photoprobes of short "arm" length, each with a substitution of either amino, iodo, or nitro at the para-position, rendering the benzophenone moiety directly radioiodinatable. Their potential for study of protein-protein interaction was assessed using the inhibitory subunit of rod cGMP phosphodiesterase (PDEgamma) and the activated transducin alphasubunit (G alpha t GTPgammaS) as a model system. These photoprobes proved to be stable at neutral pH and dithiothreitol-cleavable in addition. The PDEgamma constructs derivatized at the C-terminal positions with these probes could be readily purified, had unaltered PDEgamma functional activity, and were shown to photo-cross-link to G alpha t-GTPgammaS with an efficiency as high as 40%. Additionally, the amino benzophenone probe was radioiodinated, facilitating sensitive detection of label transfer. The uniquely combined features of these benzophenone photoprobes promise robust and flexible methods for characterization of protein-protein interaction, either by mass spectrometry when a nonradioactive label is available or by autoradiography when using radioiodinated derivatives. PMID- 15898739 TI - Effects of side chain configuration and backbone spacing on the gene delivery properties of lysine-derived cationic polymers. AB - A series of lysine-based oligomers (18 residues) that differ in side chain configuration or side chain spacing along the backbone was tested for DNA transfection activity. Although materials constructed from lysine are not the most effective polymeric transfection agents, we have chosen L-lysine-based molecules as a starting point because this system allows us to examine the functional effects of incremental changes in polycation structure. The oligomer constructed from beta(3)-homolysine (beta(3)-hLys) and that from alpha-D-lysine were superior to an alpha-L-lysine 18-mer in gene delivery assays. This improved activity is attributed to the fact that the alpha-L-peptide is a protease substrate while the other 18-mers are not. This conclusion is supported by the effects of chloroquine on transfection activity, based on the protease inhibition activity of chloroquine. To our knowledge, these results represent the first direct comparison of a D-lysine oligomer with an L-lysine oligomer in the context of gene delivery. Poly(beta(3)-hLys) was synthesized from the ring opening polymerization of the corresponding lactam. The DNA transfection ability of this polymer was compared with that of commercially available poly(L-lysine) (PLL). In each case the polymer was more active than the corresponding oligomer. PMID- 15898740 TI - Solid-phase synthesis of oligonucleotides labeled with luminescent lanthanide(III) chelates. AB - The synthesis of phosphoramidite building blocks that allow introduction of luminescent europium(III), terbium(III), dysprosium(III), and samarium(III) chelates to oligonucleotides on the solid phase is described. Several labeled oligonucleotides using these building blocks were prepared, and the photophysical properties of these bioconjugates were investigated. PMID- 15898741 TI - Identification of a UDP-glucose-binding site of human UDP-glucose dehydrogenase by photoaffinity labeling and cassette mutagenesis. AB - We have identified a UDP-glucose-binding site within human UDP-glucose dehydrogenase (hUGDH) by photoaffinity labeling with a specific probe, [(32)P]5N(3)UDP-glucose, and cassette mutagenesis using a synthetic hUGDH gene. Photolabel-containing peptides were generated by photolysis followed by tryptic digestion and isolated using the phosphopeptide isolation kit. Photolabeling of these peptides was effectively prevented by the presence of UDP-glucose during photolysis, demonstrating a selectivity of the photoprobe for the UDP-glucose binding site. Amino acid sequencing and compositional analysis identified the UDP glucose-binding site of hUGDH as the region containing the sequence, ASVGFGGSXFQK, corresponding to A268-K279 of the amino acid sequence of hUGDH. The unidentified residue, X, can be designated as a photolabeled C276 because the sequences including the cysteine residue in question have a complete identity with those of other UGDH species known. The importance of the C276 residue in the binding of UDP-glucose was further examined with mutant proteins at the C276 site. The mutagenesis at C276 has no effect on the expression of the mutants (C276G, C276K, C276E, C276L, and C276Y). Enzyme activities of the C276 mutants were not measurable under normal assay conditions, suggesting an important role for the C276 residue. No incorporation of [(32)P]5N(3)UDP-glucose was also observed for the mutants. These results indicate that C276 plays an important role for efficient binding of UDP-glucose to hUGDH. PMID- 15898742 TI - Generation of an intensely potent anthracycline by a monoclonal antibody-beta galactosidase conjugate. AB - The L49 monoclonal antibody against the p97 antigen on melanomas and carcinomas was chemically conjugated to E. coli beta-galactosidase (beta-gal), forming a largely monomeric conjugate with preserved enzymatic activity. The resulting L49 beta-gal conjugate was used to activate (N-[(4"R,S)-4"-hexyloxy-4"-(1'''-O-beta-D galactopyranosyl)butyl]daunorubicin) (1), a derivative of daunorubicin that has low cytotoxicity and high chemical stability. Addition of the conjugate to the prodrug resulted in an increase in cytotoxicity of approximately 10(5)-fold, a level of activation that is higher than any mAb-enzyme/prodrug combination yet described. Furthermore, the released drug had an IC(50) value of approximately 10 pM, making it significantly more potent than the vast majority of clinically approved anticancer drugs. The potential of this enzyme/prodrug combination for cancer therapy is discussed. PMID- 15898743 TI - A single-step method for the production of sugar hydrazides: intermediates for the chemoselective preparation of glycoconjugates. AB - The ability to selectively conjugate carbohydrate molecules to a protein is a key step in the preparation of conjugate vaccines, while facile methods for linking carbohydrates to polymers or solid surfaces to produce diagnostic probes and functional microarrays are also sought. Here, we describe a simple, single-step method of producing glycosylhydrazides from unprotected sugars, which were then linked in a controlled manner to a desired carrier, through an appropriate linker. The method was chemoselective and did not require coupling reagents, and the native pyranose form of the reducing end residue was retained. Initially, mono- and disaccharide hydrazides were produced from the corresponding reducing sugars and linked to BSA through a bifunctional linker. Final exemplification of the procedure was demonstrated by the preparation of a LewisY tetrasaccharide protein conjugate, which was recognized by a LewisY monoclonal antibody indicating the preservation of the natural conformation of the tetrasaccharide in the final construct. It is envisaged that this method will have general applicability to a variety of functionally diverse reducing sugars and provide a route to highly defined glycoconjugates, without the need for elaborate synthetic strategies. PMID- 15898744 TI - Probing for integrin alpha v beta3 binding of RGD peptides using fluorescence polarization. AB - Integrin alpha(v)beta(3) is an adhesion molecule involved in tumor invasion, angiogenesis, and metastasis. There is substantial interest in developing novel agents that bind to integrin alpha(v)beta(3). Here we report the synthesis and characterization of a fluorescent integrin alpha(v)beta(3) probe and its use in a nonradioactive, simple, sensitive fluorescence polarization (FP) assay to quantify binding to integrin alpha(v)beta(3). For assay validation, the FP assay was compared to a cell adhesion assay. In the two assays, probe binding to integrin alpha(v)beta(3) showed a similar dependence on probe concentration. The FP assay was successfully applied to measure the binding affinity to integrin alpha(v)beta(3) of several cyclic peptides containing the Arg-Gly-Asp (RGD) motif. The FP assay we describe here may be appropriate for high-throughput screening for integrin alpha(v)beta(3)-binding ligands used for anti-integrin therapy or noninvasive imaging of integrin expression. PMID- 15898745 TI - Synthesis and application of a water-soluble near-infrared dye for cancer detection using optical imaging. AB - A novel water-soluble 2-[2-(2-chloro-3-{2-[3,3-dimethyl-5-sulfo-1-(4-sulfo-butyl) 3H-indol-2-yl]-vinyl}-cyclohex-2-enylidene)-ethylidene]-3,3-dimethyl-1-(4-sulfo butyl)-2,3-dihydro-1H-indole-5-carboxylic acid (dye 2) was developed via an asymmetric approach. With an additional sulfonate group, the near-infrared feature of this dye exhibited a 2-fold increase in quantum yield compared to the previous generation. The current synthetic strategy provided a single carboxylic group as a handle for conjugation, thus allowing selectivity for bioconjugation. The stability of this dye was demonstrated by labeling peptides via solid-phase peptide chemistry. The in vivo optical imaging showed potential and broad applications of this dye in developing molecular-based beacons for cancer detection. PMID- 15898746 TI - Osmium diboride, an ultra-incompressible, hard material. AB - The need for wear- and scratch-resistant materials drives the quest for new superhard materials. In this work, we apply two design parameters to identify ultra-incompressible, superhard materials-high valence electron density and high bond covalency. Our first example of such a material is OsB2. The bulk modulus of OsB2 was measured using in situ high-pressure X-ray diffraction and was determined to be in the range of 365-395 GPa. While this value is slightly less than that of the bulk modulus of diamond, due to the anisotropic crystal structure of OsB2, the axis compressibility in the orthorhombic c-direction is less than the axis compressibility found in diamond. OsB2 also scratches the surface of a sapphire window, indicating that the hardness of OsB2 exceeds 2000 kg/mm2. PMID- 15898747 TI - 4-Connected metal-organic assemblies mediated via heterochelation and bridging of single metal ions: Kagome lattice and the M6L12 octahedron. AB - Single-metal-ion-based rigid molecular building blocks (MBBs) have been utilized to design and synthesize novel metal-organic assemblies. Reaction between In(NO3)3.2H2O and 2,5-pyridinedicarboxylic acid (2,5-H2PDC) has permitted the assembly of two supramolecular isomers, a Kagome lattice and an unprecedented M6L12 discrete octahedron. PMID- 15898748 TI - Chemically modified opals as thin permselective nanoporous membranes. AB - Thin-film opals comprising three layers of 440 nm diameter SiO2 spheres were assembled on Pt electrodes and modified with amino groups on the silica surface. Diffusion of anionic, cationic, and neutral redox species through the opals was studied by cyclic voltammetry. The chemically modified opal membranes demonstrate high molecular throughput and, at low pH, selectively block transport of a cationic redox species relative to that of anionic and neutral redox species. This permselective behavior is attributed to the electrostatic interactions that are enhanced by the tortuous pathway within the opal and by the high surface area of the chemically modified spheres. PMID- 15898749 TI - Power-law solvation dynamics in DNA over six decades in time. AB - Time-resolved Stokes shifts in a dye-containing oligonucleotide have been observed over the entire time range from 40 fs to 40 ns. The dynamics fit to a power law with a small exponent of 0.15. Similar relaxation has been seen in proteins but has not been anticipated in DNA. Distinct relaxation components due to specific subcomponents of the system, bulk water, bound water, counterions, backbone, bases, and so on, are not found. The various subcomponents may be so strongly coupled that their motions cannot be treated separately. PMID- 15898750 TI - Chiral discrimination in photochromic helicenes. AB - A photoresponsive dithienylethene derivative bearing chiral pinene-based arms underwent a stereoselective photoinduced cyclization reaction to produce greater than 98% of a single diastereomer. The magnitude of the optical rotation changed as much as Delta[alpha]373 = 8698 degrees upon alternate irradiation with 400 nm and greater than 434 nm light. PMID- 15898751 TI - Axial methionine has much less influence on reduction potentials in a CuA center than in a blue copper center. AB - The role of the highly conserved axial methionine of the purple CuA center in an engineered CuA azurin on modulating the reduction potentials of the copper center was investigated by a systematic replacement of the methionine with glutamate, aspartate, and leucine. In contrast to the same substitutions in the structurally related blue copper azurin, much smaller changes in reduction potential were observed in the CuA azurin upon replacing the methionine ligand with negatively charged Glu (-8 mV) and Asp (-5 mV) and more hydrophobic Leu (+16 mV). These findings are important in understanding the different roles of the two cupredoxins. The diamond core Cu2S2(Cys) structure of the CuA is much more resistant to variations of axial ligand interactions than the distorted tetrahedral structure of the blue copper protein. This difference may translate into a much wider range of reduction potentials (>1000 mV) for blue copper proteins that transfer electrons to a variety of partners in many different biological systems and a much narrower range of reduction potentials (<40 mV) for CuA proteins where a small difference in reduction potentials between the CuA and its redox partners is required. PMID- 15898752 TI - eta1:eta2-Alkynyl-bridged W-Si complexes: formation, structure, and reaction with acetone. AB - Reactions of (eta5-C5Me4R)(CO)2(MeCN)WMe (R = Me, Et) with HPh2SiCCtBu gave the novel alkynyl-bridged W-Si complexes, (eta5-C5Me4R)(CO)2W(mu-eta1:eta2 CCtBu)(SiPh2) (R = Me, Et), whose alkynyl ligands bridge the tungsten and silicon atoms in an eta1:eta2-coordination mode. The structures of these complexes were fully characterized, including X-ray crystallography. Treatment of (eta5 C5Me5)(CO)2W(mu-eta1:eta2-CCtBu)(SiPh2) with acetone resulted in acetone insertion into the silicon-alkynyl linkage followed by intramolecular C-H activation of the tBu group to give the chelate-type alkyl-alkene complex, (eta5 C5Me5)(CO)2W(eta1:eta2-CH2CMe2C=CHSiPh2OCMe2). PMID- 15898753 TI - Nitrosoarene-CuI complexes are intermediates in copper-catalyzed allylic amination. AB - Reactions of nitrosobenzene and N,N'-diethyl-4-nitrosoaniline with [Cu(CH3CN)4]PF6 produce novel homoleptic Cu(I)-nitrosoarene complexes, [Cu(ArNO)3]PF6, 1 (Ar = Ph) and 2 (Ar = 4-Et2NC6H4NO). The X-ray structure of 2 reveals that the copper is coordinated in a severely distorted trigonal planar geometry to the N-atom of the nitrosoarene ligand. Reactions of the PhNO complex 1 with olefins and an olefin/diene mixture provide evidence for its role as an intermediate and possibly the active nitrogen transfer agent in the Cu-catalyzed allylic amination of olefins by aryl hydroxylamines. PMID- 15898754 TI - Combining microfluidic networks and peptide arrays for multi-enzyme assays. AB - This paper reports the use of microfluidic networks (muFNs) to both prepare peptide microarrays and carry out label-free enzyme assays on self-assembled monolayers (SAMs) of alkanethiolates on gold. A poly(dimethylsiloxane) (PDMS) stamp fabricated with microchannels is used to immobilize a linear array of cysteine-terminated peptides onto SAMs presenting maleimide groups. The stamp is then reapplied to the SAM in a perpendicular direction to introduce enzyme solutions so that each solution can interact with an identical linear array of immobilized peptides. The muFNs enable multiple enzyme-substrate interactions to be simultaneously evaluated at a submicroliter scale, while the use of SAMs enables the use of MALDI mass spectrometry (MS) to analyze the enzyme activities. This paper demonstrates applications of this system for assaying multiple kinases and for profiling the activities of kinases and phosphatases in human K562 cell extracts. The combination of muFN, SAMs, and MS detection provides a flexible platform for assaying enzyme activities in biological samples. PMID- 15898755 TI - Two-photon absorption at telecommunications wavelengths in a dipolar chromophore with a pyrrole auxiliary donor and thiazole auxiliary acceptor. AB - Three new dipolar chromophores based on a diaklyaminophenyl donor, a pyrrole auxiliary donor, a thiazole auxiliary acceptor, and strong heterocyclic acceptors have been synthesized. For one of these compounds we have measured a very large non-degenerate two-photon cross section of ca. 1500 GM in the near-IR telecommunications range using a pump-probe technique. Calculations indicate the cross section for degenerate two-photon absorption is likely to be ca. 60% of this value. PMID- 15898756 TI - Catalytic enantioselective thioester aldol reactions that are compatible with protic functional groups. AB - This communication reports highly enantioselective and diastereoselective methyl malonic acid half thioester (MeMAHT) aldol reactions that are compatible with protic functional groups and enolizable aldehydes, affording syn S-phenyl thiopropionates. PMID- 15898757 TI - 14-Hydroxylation of opiates: catalytic direct autoxidation of codeinone to 14 hydroxycodeinone. AB - Codeinone (3) was efficiently and directly converted to 14-hydroxycodeinone (1) by catalytic air oxidation in aqueous solution. A number of simple manganese and copper salts were identified to be effective catalysts, including MnSO4, KMnO4, and CuSO4. An appropriate reducing agent, such as sodium thiosulfate, is required in the reaction mixture presumably for the reduction of a detrimental peroxide intermediate. This discovery allows the more abundant codeine to be employed as the starting material for the synthesis of 14-hydroxylated opiate drugs without recourse to a thebaine-like intermediate. These discoveries were inspired from our study of microbial transformation of codeine to 14-hydroxycodeine by Mycobacterium neoaurum, where we found the actual 14-hydroxylation step is a chemical reaction rather than an enzymatic reaction, as previously believed. PMID- 15898758 TI - Catalytic and asymmetric vinylogous mukaiyama reactions on aliphatic ketones: formal asymmetric synthesis of taurospongin a. AB - Catalytic asymmetric vinylogous Mukaiyama reactions on ketones, leading to the formation of alpha,beta-unsaturated lactones with tertiary alcohols, have been described (11 examples, up to 93% ee). This methodology has been applied in a formal enantioselective synthesis of taurospongin A (12 steps, 6% overall yield). PMID- 15898759 TI - Total synthesis of mycalamide A. AB - This communication describes a concise and efficient total synthesis of mycalamide A by the convergent coupling of pederic acid unit with the mycalamine unit. The left-half, (+)-7-benzoylpederic acid, was synthesized from (2R,3R)-3 methylpent-4-en-2-ol in seven steps and 34.6% overall yield through a route that features a one-step Pd(II)-catalyzed tandem Wacker/Heck cyclization reaction to prepare the tetrahydropyran ring system. The right-half, the mycalamine unit, was synthesized in 21 steps and 10.5% overall yield from diethyl d-tartrate. Effective, stereoselective methods were developed for the assembly of the two parts to yield either mycalamide A or C(10)-epi-mycalamide A. PMID- 15898760 TI - 2D correlation analysis of the continuum in single molecule surface enhanced Raman spectroscopy. AB - The role and the nature of the continuum in Surface Enhanced Raman Spectroscopy (SERS) are unclear. Here, two-dimensional (2D) covariance and correlation analysis is applied to single molecule SERS spectra on silver colloids with and without rhodamine 6G (native colloid). The resulting 2D covariance and correlation maps show that the sharp molecular Raman peaks from rhodamine 6G and the molecule responsible for the SERS peaks from the native colloid are correlated to different continua even though both continua are present in each data set. This suggests that two distinct active sites on the silver colloids produce the two different continua, and that each site has some molecular specificity. PMID- 15898761 TI - Fixation of both O2 and CO2 from air by a crystalline palladium complex bearing N heterocyclic carbene ligands. AB - Crystals of the two-coordinate palladium(0) complex 1 bearing the new N heterocyclic carbene ligand, ITmt, directly and rapidly fixed both O2 and CO2 from air to produce the corresponding palladium(II) peroxocarbonate complex 2. The present reaction consists of dioxygenation of the palladium(0) complex 1 to the palladium(II) peroxo complex 3 and the subsequent CO2 insertion to produce the peroxocarbonate complex 2. Reaction of the crystals of 1 with air was monitored by microscopic IR spectroscopy to confirm the sequence of the two-step solid-state reaction. The unique reactivity of solid 1 toward air was explained in terms of the structural features of the carbene ligand, ITmt. PMID- 15898762 TI - Hydrophobically assisted switching phase synthesis: the flexible combination of solid-phase and solution-phase reactions employed for oligosaccharide preparation. AB - Hydrophobically assisted switching phase (HASP) synthesis is a concept that allows the choice between the advantages of solid-supported chemistry and those of solution-phase synthesis. Starting from the examination of adsorption and desorption properties of hydrophobic molecules to and from reversed-phase silica, we designed a dilipid as a quantitative and fully reversible HASP anchor, permitting final product release. The utility of this new tool in synthetic organic chemistry was demonstrated on oligosaccharide preparation. The synthesis of a pentarhamnoside was accomplished by repetitive glycosylation reactions. Glycosylations were conducted preferably in solution, whereas all protecting group manipulations were performed on solid support. Without the need for chromatographic purification of intermediates, the HASP system furnished the final product after 12 linear steps with average yields of 94% per step at a scale of 0.1 mmol, thus overcoming several of the limitations encountered in the solid-phase synthesis of complex carbohydrates. PMID- 15898763 TI - Sol-gel reaction of porphyrin-based superstructures in the organogel phase: creation of mechanically reinforced porphyrin hybrids. AB - We have demonstrated that a one-dimensional molecular assembly created by an H aggregated porphyrin.Cu(II) stack can be immobilized, without a morphological change, by sol-gel polycondensation of the peripheral triethoxysilyl groups. The resultant gel prepared according to this flowchart has gained a very high thermal stability as well as a unique mechanical strength. PMID- 15898764 TI - Quantum control of a chiral molecular motor driven by laser pulses. AB - "Molecular motors or machines" are one of the hot subjects in chemistry because they play an important role in molecular devices. We have theoretically demonstrated that unidirectional rotations of a chiral molecular motor can be driven by using tailored linearly polarized laser pulses. The findings obtained here serve as a theoretical basis for control of functions such as gearing or acceleration of molecular motors. PMID- 15898765 TI - Ionic liquid-in-oil microemulsions. AB - Phase stability and small-angle neutron scattering (SANS) data show that surfactant-stabilized nanodomains of a typical ionic liquid (1-butyl-3 methylimidazolium tetrafluoroborate [bmim][BF4]) may be dispersed by the nonionic surfactant Triton-X100 in cyclohexane. Analyses of these SANS data are consistent with the formation of ionic liquid-in-oil microemulsion droplets. PMID- 15898766 TI - Syntheses of shell cross-linked micelles using acidic ABC triblock copolymers and their application as pH-responsive particulate emulsifiers. AB - The use of new shell cross-linked micelles as pH-responsive particulate emulsifiers is described for the first time. 1-Undecanol-in-water emulsions of 18 mum diameter are formed at pH 8, whereas complete demulsification occurs at pH 2. PMID- 15898767 TI - Dual species emission from single polyfluorene molecules: signatures of stress induced planarization of single polymer chains. AB - Single chains of the conjugated polymer polyfluorene are shown to exist in two distinct conformational arrangements. Planarization of the chain to the single molecule beta-phase leads to a red shift in the emission and a strong modification of the vibronic progression. Most importantly, this structural rearrangement dramatically affects the photophysical stability on the single molecule level. Single molecule beta-phase emission displays a vastly improved lifetime and much less noise on both the emission intensity and the spectral position. In the absence of signatures of multichromophoric emission on the single molecule level, we propose that the effective conjugation length accounts for most of the physical chain length of the beta-phase. PMID- 15898768 TI - Pd(II)-catalyzed intermolecular 1,2-diamination of conjugated dienes. AB - A Pd(II) chloride precatalyst, in the presence of 1 equiv of benzoquinone, effects highly efficient, regioselective 1,2-diamination of 1,3-dienes using dialkyl ureas under mild conditions. There is no requirement for a large excess of diene. PMID- 15898769 TI - Escherichia coli quinolinate synthetase does indeed harbor a [4Fe-4S] cluster. AB - Quinolinic acid is an intermediate in the biosynthesis of nicotinamide-containing redox cofactors. The ultimate step in the formation of quinolinic acid in prokaryotes is the condensation of iminosuccinate and dihydroxyacetone phosphate, which is catalyzed by the product of the nadA gene in Escherichia coli. A combination of UV-vis, Mossbauer, and EPR spectroscopies, along with analytical methods for the determination of iron and sulfide, demonstrates for the first time that anaerobically purified quinolinate synthetase (NadA) from E. coli contains one [4Fe-4S] cluster per polypeptide. The protein is active, catalyzing the formation of quinolinic acid with a Vmax [ET]-1 of 0.01 s-1. PMID- 15898770 TI - Persistent (amino)(silyl)carbenes. AB - (Amino)(silyl)carbenes, prepared by substitution reactions at a carbene center, can survive several days at 0 degrees C. These species are not push-pull carbenes as their phosphino analogues and therefore are excellent ligands for transition metal centers. PMID- 15898771 TI - Structural understanding of a molecular material that is accessed only by a solid state desolvation process: the scope of modern powder X-ray diffraction techniques. AB - Many molecular materials cannot be prepared as a "pure" (nonsolvate) crystalline phase by conventional crystal growth from solution due to the facile formation of solvate structures. In such cases, it may be possible to obtain the pure phase by a solid-state desolvation process, although such processes are generally associated with loss of crystal integrity, yielding a microcrystalline powder of the pure phase. This paper demonstrates the utility of modern powder X-ray diffraction techniques for obtaining structural understanding in such cases, focusing on a particular member of a structural family that is of wider relevance within the context of crystal engineering and design. PMID- 15898772 TI - Magnetic nanotubes for magnetic-field-assisted bioseparation, biointeraction, and drug delivery. AB - Tubular structure of nanoparticles is highly attractive due to their structural attributes, such as the distinctive inner and outer surfaces, over conventional spherical nanoparticles. Inner voids can be used for capturing, concentrating, and releasing species ranging in size from large proteins to small molecules. Distinctive outer surfaces can be differentially functionalized with environment friendly and/or probe molecules to a specific target. Magnetic particles have been extensively studied in the field of biomedical and biotechnological applications, including drug delivery, biosensors, chemical and biochemical separation and concentration of trace amounts of specific targets, and contrast enhancement in magnetic resonance imaging (MRI). Therefore, by combining the attractive tubular structure with magnetic property, the magnetic nanotube (MNT) can be an ideal candidate for the multifunctional nanomaterial toward biomedical applications, such as targeting drug delivery with MRI capability. Here, we successfully synthesized magnetic silica-iron oxide composite nanotubes and demonstrated the magnetic-field-assisted chemical and biochemical separations, immunobinding, and drug delivery. PMID- 15898773 TI - A class of ruthenium(II) catalyst for asymmetric transfer hydrogenations of ketones. AB - Ruthenium dimer 6 (readily available in two steps from TsDPEN) is converted directly to monomeric asymmetric transfer hydrogenation catalyst 3 in situ under the conditions employed for ketone reduction. Catalyst 3 is a significantly more active catalyst for this application than the untethered derivative, exhibits higher enantioselectivities across a range of substrates, and appears to be highly stable to the reaction conditions. It is active at loadings of as low as 0.01 mol %, and reductions at the 0.1 mol % level are complete within 20 min at 80 degrees C without significant loss of enantioselectivity. PMID- 15898774 TI - Highly enantioselective and regioselective nickel-catalyzed coupling of allenes, aldehydes, and silanes. AB - A complex derived from Ni(cod)2 and NHC-IPr catalyzes a three-component coupling reaction involving allenes, aldehydes, and organosilanes and transfers the axial chirality of the allene to a stereogenic center in the product with very high fidelity. An unexpected regioselectivity is observed; favored are allylic rather than homoallylic alcohol derivatives, corresponding to the unusual process of coupling two electrophilic atoms: the allene sp and aldehyde carbon atoms. In all cases, high enantioselectivity, high Z/E selectivity, and, with differentially substituted allenes, high site selectivity are observed. This transformation represents the first enantioselective multicomponent coupling process of allenes. PMID- 15898775 TI - What is the effective charge of TGA-stabilized CdTe nanocolloids? AB - The surface charge of semiconductor nanoparticles, Q, is an important parameter which determines their electrokinetic behavior, stability in water and polar solvents, functions of optical and electronic devices, self-assembly properties, and interactions with cell membranes. We have developed a simple method for quantitative determination of Q in their native aqueous environment. The method does not require the knowledge of exact atomic structure or make assumptions about effects of drying on charge distribution. The method is based on titration of nanoparticle dispersion with a solution of oppositely charged polyelectrolyte. The point of complete neutralization is recognized as an inflection point on the dependence of fluorescence intensity on the amount of polyelectrolyte added. Thioglycolic acid-stabilized CdTe nanoparticles 2 nm in diameter were found to carry an average Q from -2.6 to -5.5 for pH 7.5 to 10, respectively. This charge is found to be smaller than that calculated theoretically for an analogous structure (i.e., Q = -8), presumably due to adsorption of Cd(2+) ions on the stabilizer shell and on Te atoms with unsaturated valence located on the side planes of CdTe tetrahedrons. PMID- 15898776 TI - Radical addition to "cation pool". Reverse process of radical cation fragmentation. AB - The reaction of an N-acyliminium ion with an alkyl iodide and hexabutyldistannane took place to give the alkylation product. A mechanism involving the addition of an alkyl radical to an N-acyliminium ion to produce the corresponding radical cation has been suggested. PMID- 15898777 TI - Low-temperature polyol synthesis of AuCuSn2 and AuNiSn2: using solution chemistry to access ternary intermetallic compounds as nanocrystals. AB - Ternary intermetallic compounds, which possess a wide variety of important properties with both academic and technological relevance, are typically synthesized using traditional high-temperature methods. Here, we demonstrate that the polyol method, which is used extensively to synthesize nanocrystals and nanocrystalline powders of metals and simple binary compounds, serves as an effective low-temperature exploratory medium for synthesizing new ordered ternary intermetallics as nanocrystals. Accordingly, we describe the synthesis and structural characterization of AuCuSn2 and AuNiSn2, which adopt an ordered NiAs type superstructure that is not observed using equilibrium synthetic methods. AuCuSn2 forms in solution of 120 degrees C as well-formed nanocrystals, and the ordered phase is stable up to 450 degrees C. AuNiSn2 behaves similarly to AuCuSn2. PMID- 15898778 TI - Dithiocarbamate assembly on gold. AB - Au surfaces are functionalized by stable dithiocarbamate ligands when exposed to carbon disulfide and secondary amines. The adsorbed dithiocarbamates are robust under a wide pH range and can resist displacement by other chemisorptive surfactants, providing an attractive method for conjugating sensitive molecules onto metal surfaces. PMID- 15898779 TI - Oxidative C-H activation/C-C bond forming reactions: synthetic scope and mechanistic insights. AB - This paper describes a new palladium-catalyzed method for C-H activation/carbon carbon bond formation with hypervalent iodine arylating agents. This transformation has been applied to a variety of arene and benzylic substrates containing different directing groups (pyridines, quinolines, oxazolidinones, and amides) and proceeds with high levels of regiocontrol. Mechanistic experiments provide preliminary evidence in support of an unusual mechanism for this transformation involving a Pd(II)/Pd(IV) catalytic cycle. PMID- 15898780 TI - Understanding the role of oxidative etching in the polyol synthesis of Pd nanoparticles with uniform shape and size. AB - Uniform cubooctahedral nanoparticles of Pd were synthesized using a modified polyol process. Like the silver system, the Pd nanoparticles underwent dissolution due to oxidative etching by Cl-/O2. In the early stage of etching, the twinned particles in a sample were selectively removed to leave behind uniform, single-crystal cubooctahedra. If the etching was allowed to continue, the single-crystal cubooctahedra were also attacked and partially dissolved. Understanding the role of oxidative etching is critical to the achievement of both uniform shape and size. PMID- 15898781 TI - Isocomplestatin: total synthesis and stereochemical revision. AB - A Pd-mediated method for preparation of the strained macrocyclic moiety of complestatins is disclosed. Through stereoselective synthesis of model macrocycles and the S atropisomer of complestatin, the stereochemical identity of the anti-HIV agent complestatin is established. Investigations described herein illustrate that the compound previously reported as isocomplestatin is the same as complestatin. Thus, the S atropisomer of complestatin is the true isocomplestatin and has not been isolated as a natural product. PMID- 15898782 TI - Probing the interior of peptide amphiphile supramolecular aggregates. AB - We present a study of the aqueous solvation within self-assembled structures formed from peptide amphiphiles. We have placed tryptophan and pyrene chromophores onto the peptide backbone to enable spectroscopic examinations of the interior of the resulting supramolecular objects. Self-assembly constrains the chromophores to a defined location within an aggregate, and they experience differing degrees of quencher penetration reflective of their depth within the nanostructure. Tryptophan fluorescence indicates that the interiors remain well solvated, suggesting that the supramolecular aggregates maintain high degrees of free volume. The Stern-Volmer quenching constants and the fractional accessibility (of covalently bound pyrene) progressively increase as the chromophore is placed closer to the aggregate exterior. Furthermore, these aggregates encourage chromophore uptake from aqueous solution as evidenced by the solubilization of free pyrene chromophores. Our findings demonstrate that covalently bound fluorophores within an aggregate can interact with the external environment. Studies with small molecular probes indicate that these self assembled architectures may represent viable vehicles to sequester hydrophobic, insoluble organic molecules (within the interior) and to present signaling protein epitopes to cells (on the periphery). PMID- 15898783 TI - Cytosine catalysis of nitrosative guanine deamination and interstrand cross-link formation. AB - Effects are discussed of the anisotropic DNA environment on nitrosative guanine deamination based on results of an ab initio study of the aggregate 3 formed by guaninediazonium ion 1 and cytosine 2. Within 3, the protonation of 2 by 1 is fast and exothermic and forms 6, an aggregate between betaine 4 (2-diazonium-9H purin-6-olate) and cytosinium ion 5. Electronic structure analysis of 4 shows that this betaine is not mesoionic; only the negative charge is delocalized in the pi-system while the positive charge resides in the sigma-system. Potential energy surface exploration shows that both dediazoniation and ring-opening of betaine 4 in aggregate 6 are fast and exothermic and lead irreversibly to E-11, the aggregate between (E)-5-cyanoimino-4-oxomethylene-4,5-dihydroimidazole E-10 and 5. The computed pair binding energies for 3, 6, and E-11 greatly exceed the GC pair binding energy. While 1 can be a highly reactive intermediate in reactions of the "free nucleobase" (or its nucleoside and nucleotide), the cyanoimine 10 emerges as the key intermediate in nitrosative guanine deamination in ds-DNA and ds-oligonucleotides. In essence, the complementary nucleobase cytosine provides base catalysis and switches the sequence of deprotonation and dediazoniation. It is argued that this environment-induced switch causes entirely different reaction paths to products as compared to the respective "free nucleobase" chemistry, and the complete consistency is demonstrated of this mechanistic model with all known experimental results. Products might form directly from 10 by addition and ring closure, or their formation might involve water catalysis via 5-cyanoamino-4-imidazolecarboxylic acid 12 and/or 5 carbodiimidyl-4-imidazolecarboxylic acid 13. The pyrimidine ring-opened intermediates 10, 12, and 13 can account for the formations of xanthosine, the pH dependency and the environment dependency of oxanosine formation, the formation of the classical cross-link dG(N(2)())-to-dG(C2), including the known sequence specificity of its formation, and the formation of the structure-isomeric cross link dG(N1)-to-dG(C2). PMID- 15898784 TI - Aziridine-2-carboxylic acid-containing peptides: application to solution- and solid-phase convergent site-selective peptide modification. AB - The development of a method for site- and stereoselective peptide modification using aziridine-2-carboxylic acid-containing peptides is described. A solid-phase peptide synthesis methodology that allows for the rapid generation of peptides incorporating the aziridine residue has been developed. The unique electrophilic nature of this nonproteinogenic amino acid allows for site-selective conjugation with various thiol nucleophiles, such as anomeric carbohydrate thiols, farnesyl thiol, and biochemical tags, both in solution and on solid support. This strategy, combined with native chemical ligation, provides convergent and rapid access to complex thioglycoconjugates. PMID- 15898785 TI - Reactions of the peroxo intermediate of soluble methane monooxygenase hydroxylase with ethers. AB - Soluble methane monooxygenase (sMMO) isolated from Methylococcus capsulatus (Bath) utilizes a carboxylate-bridged diiron center and dioxygen to catalyze the conversion of methane to methanol. Previous studies revealed that a di(mu oxo)diiron(IV) intermediate termed Q is responsible for the catalytic activity with hydrocarbons. In addition, the peroxodiiron(III) intermediate (H(peroxo)) that precedes Q formation in the catalytic cycle has been demonstrated to react with propylene, but its reactivity has not been extensively investigated. Given the burgeoning interest in the existence of multiple oxidants in metalloenzymes, a more exhaustive study of the reactivity of H(peroxo) was undertaken. The kinetics of single turnover reactions of the two intermediates with ethyl vinyl ether and diethyl ether were monitored by single- and double-mixing stopped-flow optical spectroscopy. For both substrates, the rate constants for reaction with H(peroxo) are greater than those for Q. An analytical model for explaining the transient kinetics is described and used successfully to fit the observed data. Activation parameters were determined through temperature-dependent studies, and the kinetic isotope effects for the reactions with diethyl ether were measured. The rate constants indicate that H(peroxo) is a more electrophilic oxidant than Q. We propose that H(peroxo) reacts via two-electron transfer mechanisms, and that Q reacts by single-electron transfer steps. PMID- 15898786 TI - Molecular recognition of saccharides by proteins. Insights on the origin of the carbohydrate-aromatic interactions. AB - The existence of stabilizing carbohydrate-aromatic interactions is demonstrated from both the theoretical and experimental viewpoints. The geometry of experimentally based galactose-lectin complexes has been properly accounted for by using a MP2/6-31G(d,p) level of theory and by considering a counterpoise correction during optimization. In this case, the stabilizing interaction energy of the fucose-benzene complex amounts to 3.0 kcal/mol. The theoretical results obtained herein indicate that the carbohydrate-aromatic interactions are stabilizing interactions with an important dispersive component and that electronic density between the sugar hydrogens and the aromatic ring indeed exists, thus giving rise to three so-called nonconventional hydrogen bonds. Experimental evidence of the intrinsic tendency of aromatic moieties to interact with certain sugars has also been shown by simple NMR experiments in water solution. Benzene and phenol specifically interact with the clusters of C-H bonds of the alpha face of methyl beta-galactoside, without requiring the well-defined three-dimensional shape provided by a protein receptor, therefore resembling the molecular recognition features that are frequently observed in many carbohydrate protein complexes. PMID- 15898787 TI - Measurement of ribose carbon chemical shift tensors for A-form RNA by liquid crystal NMR spectroscopy. AB - Incomplete motional averaging of chemical shift anisotropy upon weak alignment of nucleic acids and proteins in a magnetic field results in small changes in chemical shift. Knowledge of nucleus-specific chemical shift (CS) tensor magnitudes and orientations is necessary to take full advantage of these measurements in biomolecular structure determination. We report the determination by liquid crystal NMR of the CS tensors for all ribose carbons in A-form helical RNA, using a series of novel 3D NMR pulse sequences for accurate and resolved measurement of the ribose (13)C chemical shifts. The orientation of the riboses relative to the rhombic alignment tensor of the molecule studied, a stem-loop sequence corresponding to helix-35 of 23S rRNA, is known from an extensive set of residual dipolar couplings (RDC), previously used to refine its structure. Singular-value-decomposition fits of the chemical shift changes to this structure, or alternatively to a database of helical RNA X-ray structures, provide the CS tensor for each type of carbon. Quantum chemical calculations complement the experimental results and confirm that the most shielded tensor component lies approximately along the local carbon-oxygen bond axis in all cases and that shielding anisotropy for C3' and C4' is much larger than for C1' and C2', with C5' being intermediate. PMID- 15898788 TI - Ionic thermotropic liquid crystal dendrimers. AB - The synthesis and characterization of two families of ionic liquid crystal dendrimers consisting of the ammonium salts of the commercially available poly(amidoamine) (G = 0-5) and poly(propylene imine) (G = 1-5) dendrimers and three long-chain carboxylic acids are reported. The liquid crystalline behavior was investigated by means of differential scanning calorimetry, polarizing light optical microscopy, and X-ray diffractometry. The thermal stability of the ionic materials was further studied by NMR. Most of the dendrimers show lamellar mesomorphism, and two of them exhibit columnar mesomorphism. On the basis of the experimental results, we propose models both at the molecular level and in the mesophase for all the materials. PMID- 15898789 TI - PEG- and peptide-grafted aliphatic polyesters by click chemistry. AB - Novel aliphatic polyesters with pendent acetylene groups were prepared by controlled ring-opening polymerization and subsequently used for grafting poly(ethylene glycol) and oligopeptide moieties by the Cu(I)-catalyzed addition of azides and alkynes, a type of "click" chemistry. These aliphatic polyesters possess an acetylene graft density that can be tailored by ring-opening copolymerization of alpha-propargyl-delta-valerolactone (1) with epsilon caprolactone. Since the mild conditions associated with the click reaction are shown to be compatible with the polyester backbone, this method is a generally useful means for grafting numerous types of functionality onto aliphatic polyesters. The amphiphilic graft polyesters prepared in this study are shown to be biocompatible by in vitro cytotoxicity evaluation, suggesting their suitability for a range of biomaterial applications. PMID- 15898790 TI - Intermolecular electron-transfer mechanisms via quantitative structures and ion pair equilibria for self-exchange of anionic (dinitrobenzenide) donors. AB - Definitive X-ray structures of "separated" versus "contact" ion pairs, together with their spectral (UV-NIR, ESR) characterizations, provide the quantitative basis for evaluating the complex equilibria and intrinsic (self-exchange) electron-transfer rates for the potassium salts of p-dinitrobenzene radical anion (DNB(-)). Three principal types of ion pairs, K(L)(+)DNB(-), are designated as Classes S, M, and C via the specific ligation of K(+) with different macrocyclic polyether ligands (L). For Class S, the self-exchange rate constant for the separated ion pair (SIP) is essentially the same as that of the "free" anion, and we conclude that dinitrobenzenide reactivity is unaffected when the interionic distance in the separated ion pair is r(SIP) > or =6 Angstroms. For Class M, the dynamic equilibrium between the contact ion pair (with r(CIP) = 2.7 Angstroms) and its separated ion pair is quantitatively evaluated, and the rather minor fraction of SIP is nonetheless the principal contributor to the overall electron transfer kinetics. For Class C, the SIP rate is limited by the slow rate of CIP right arrow over left arrow SIP interconversion, and the self-exchange proceeds via the contact ion pair by default. Theoretically, the electron-transfer rate constant for the separated ion pair is well-accommodated by the Marcus/Sutin two state formulation when the precursor in Scheme 2 is identified as the "separated" inner-sphere complex (IS(SIP)) of cofacial DNB(-)/DNB dyads. By contrast, the significantly slower rate of self-exchange via the contact ion pair requires an associative mechanism (Scheme 3) in which the electron-transfer rate is strongly governed by cationic mobility of K(L)(+) within the "contact" precursor complex (IS(CIP)) according to the kinetics in Scheme 4. PMID- 15898791 TI - Targeting the tumor-associated folate receptor with an 111In-DTPA conjugate of pteroic acid. AB - The cell membrane folate receptor is a potential molecular target for tumor selective drug delivery. To probe structural requirements for folate receptor targeting with low molecular weight radiometal chelates, specifically the role of the amino acid fragment of folic acid (pteroylglutamic acid) in mediating targeting selectivity, the amide-linked conjugate pteroyl NHCH(2)CH(2)OCH(2)CH(2)OCH(2)CH(2)NH-DTPA was prepared by a three-step procedure from pteroic acid, 2,2'-(ethylenedioxy)-bis(ethylamine), and t-Bu-protected DTPA. This conjugate, 1-{2-[2-[(2-(biscarboxymethyl-amino)ethyl)-carboxymethyl amino]ethyl]-carboxymethyl-amino}-acetylamino-3,6-dioxa-8-pteroylamino-octane (1), was employed for synthesis of the corresponding (111)In(III) radiopharmaceutical. Following intravenous administration to athymic mice, the (111)In complex of 1 was found to selectively localize in folate receptor positive human KB tumor xenografts and to afford prolonged tumor retention of the (111)In radiolabel (5.4 +/- 0.8, 5.6 +/- 1.1, and 3.6 +/- 0.6% of the injected dose per gram of tumor at 1, 4, and 24 h, respectively). The observed tumor localization was effectively blocked by co-administration of folic acid with the (111)In-1 complex, consistent with a folate receptor-mediated targeting process. In control studies, tumor targeting with this pteroic acid conjugate appears as effective as that seen using (111)In-DTPA-folate, a radiopharmaceutical that has progressed to clinical trials for detection of folate receptor-expressing gynecological tumors. PMID- 15898793 TI - Solvatochromism of distyrylbenzene pairs bound together by [2.2]paracyclophane: evidence for a polarizable "through-space" delocalized state. AB - A series of compounds were designed and synthesized to examine how through-space and through-bond electron delocalization respond to solvent effects. The general strategy involves the study of "dimers" of the distyrylbenzene chromophore held in close proximity by the [2.2]paracyclophane core and a systematic dissection of the chromophore into components with through-space and through-bond electronic delocalization. Steady state and time-resolved fluorescence spectroscopy in a range of solvents reveals a red-shift in emission and an increase in the intrinsic fluorescence lifetime for the emitting state in polar solvents when donor substituents are absent. We propose that through-space delocalization across the [2.2]paracyclophane core is more polarizable in the excited state, relative to the through-bond (distyrylbenzene based) excited state. When strong donors are attached to the distyrylbenzene chromophore, the charge transfer character of the distyrylbenzene-based excited state dominates fluorescence properties. PMID- 15898792 TI - Kinetic analysis of an efficient DNA-dependent TNA polymerase. AB - alpha-l-Threofuranosyl nucleoside triphosphates (tNTPs) are tetrafuranose nucleoside derivatives and potential progenitors of present-day beta-d-2' deoxyribofuranosyl nucleoside triphosphates (dNTPs). Therminator DNA polymerase, a variant of the 9 degrees N DNA polymerase, is an efficient DNA-directed threosyl nucleic acid (TNA) polymerase. Here we report a detailed kinetic comparison of Therminator-catalyzed TNA and DNA syntheses. We examined the rate of single-nucleotide incorporation for all four tNTPs and dNTPs from a DNA primer template complex and carried out parallel experiments with a chimeric DNA-TNA primer-DNA template containing five TNA residues at the primer 3'-terminus. Remarkably, no drop in the rate of TNA incorporation was observed in comparing the DNA-TNA primer to the all-DNA primer, suggesting that few primer-enzyme contacts are lost with a TNA primer. Moreover, comparison of the catalytic efficiency of TNA synthesis relative to DNA synthesis at the downstream positions reveals a difference of no greater than 5-fold in favor of the natural DNA substrate. This disparity becomes negligible when the TNA synthesis reaction mixture is supplemented with 1.25 mM MnCl(2). These results indicate that Therminator DNA polymerase can recognize both a TNA primer and tNTP substrates and is an effective catalyst of TNA polymerization despite changes in the geometry of the reactants. PMID- 15898794 TI - Mechanism of enyne metathesis catalyzed by Grubbs ruthenium-carbene complexes: a DFT study. AB - The complete catalytic cycle of the reaction of alkenes and alkynes to dienes by Grubbs ruthenium carbene complexes has been modeled at the B3LYP/LACV3P**+//B3LYP/LACVP level of theory. The core structures of the substrates and the catalyst were used as models, namely, ethene, ethyne, hept-1 en-6-yne, (Me(3)P)(2)Cl(2)Ru=CH(2), and [C(2)H(4)(NMe)(2)C](Me(3)P)Cl(2)Ru=CH(2). Insight into the electronically most preferred mechanistic pathways was gained for both intermolecular as well as for intramolecular enyne metathesis. Alkene metathesis is predicted to proceed fast and reversible, while the insertion of the alkyne substrate is slower, irreversible, and kinetically regioselectivity determining. Ruthenacyclobut-2-ene structures do not exist as local minima in the catalytic cycle. Instead, vinylcarbene complexes are formed directly. The alkyne insertion step and the cycloreversion of 2-vinyl ruthenacyclobutanes feature comparable predicted overall barriers in intermolecular enyne metathesis. For intramolecular enyne metathesis, a noncyclic alkene fragment of the enyne substrate is first incorporated into the Grubbs catalyst by an alkene metathesis reaction. The subsequent insertion of the alkyne fragment then proceeds intramolecularly. Alkene association, cycloaddition, and cycloreversion to the diene product complex close the catalytic cycle. Rate enhancement by an ethene atmosphere (Mori's conditions) originates from a constantly higher overall alkene concentration that is necessary for the rate-limiting [2 + 2] cycloreversion step to the diene product complex. PMID- 15898795 TI - Unexpected stability of aryl beta-N-acetylneuraminides in neutral solution: biological implications for sialyl transfer reactions. AB - A reagent panel comprised of seven aryl beta-D-N-acetylneuraminides was synthesized and then used to probe the mechanisms of nonenzymatic hydrolysis. These reactions proceeded via four independent pathways: (1) acid-catalyzed hydrolysis of the neutral molecule; (2) acid-catalyzed hydrolysis of the anionic form, or its kinetic equivalent spontaneous hydrolysis of the neutral form; (3) spontaneous hydrolysis of the anionic form; and (4) a base-promoted pathway. The pH-independent spontaneous hydrolysis of 4-nitrophenyl alpha-D-N acetylneuraminide (5) occurs at a rate that is over 100 times faster than that of the corresponding reaction of 4-nitrophenyl beta-D-N-acetylneuraminide (4a). Spontaneous hydrolyses of four aryl beta-D-N-acetylneuraminides displayed a beta(lg) value of -1.24 +/- 0.16 (pH = 8.1, T = 100 degrees C), and at a pH value of 1.0 (50 degrees C), all seven panel members gave a beta(lg) value of 0.14 +/- 0.08. The aqueous ethanolyses of 4a and 5 gave similar products and displayed sensitivity parameters (m) in a standard Winstein-Grunwald analysis of -0.04 +/- 0.01 and +0.23 +/- 0.02, respectively. These results, plus the activation parameters calculated for the spontaneous hydrolyses of the anionic forms of 5 (DeltaH() = 116 +/- 2 kJ mol(-1) and DeltaS = 27 +/- 4 J mol(-1) K(-1)) and 4a (DeltaH = 138 +/- 3 kJ mol(-1) and DeltaS = 59 +/- 8 J mol(-1) K(-1)), are inconsistent with anomeric carboxylate assistance occurring during the hydrolysis reactions, and the likely cause for the enhanced reactivity of 5 in comparison to that of 4a is an increase in ground-state steric strain. PMID- 15898796 TI - The gas-phase acidity of 2(3H)-oxepinone: a step toward an experimental heat of formation for the 2-oxepinoxy radical. AB - In an effort to gain further insight into the oxidation of the phenyl radical, this contribution details the first of three experiments designed to establish the heat of formation of the 2-oxepinoxy radical. We report here the synthesis of the previously unknown 2(7H)-oxepinone (12a) and 2(3H)-oxepinone (12b). We have determined the gas-phase acidity (Delta(acid)H(298)) of 12b by means of a bracketing study employing a flowing afterglow apparatus with quadrupole mass spectrometric detection. In this experiment, compound 12b was reacted in the gas phase with a series of bases of varying strength. A proton-transfer reaction was observed when 12b was reacted with t-BuS(-), but not when 12b was reacted with HS(-). We conclude that the gas-phase acidity of 12b lies between those of t-BuSH and H(2)S, and it is thereby assigned a value of Delta(acid)H(298) = 352 +/- 2 kcal/mol. Additional support for this value was found by performing the reverse reactions (i.e. the 2-oxepinoxy anion (15a) was reacted with proton sources of differing acidities). Anion 15a underwent a proton-transfer reaction with H(2)S but not with t-BuSH, in agreement with the results from the forward reactions. The experimental value of the gas-phase acidity agrees well with those from DFT calculations, which predicted Delta(acid)H(298) = 348.9 kcal/mol at the B3LYP/6 31+G(d) level and 349.2 kcal/mol at the B3LYP/aug-cc-pVTZ level. PMID- 15898797 TI - Facile preparation of various heteroaromatic compounds via azatitanacyclopentadiene intermediates. AB - Coupling of acetylene, nitrile, and a titanium reagent, Ti(O-i-Pr)(4)/2 i-PrMgCl, generated new azatitanacyclopentadienes in a highly regioselective manner. Their subsequent reaction with sulfonylacetylene afforded pyridyltitanium compounds, which, upon reaction with electrophiles, gave substituted pyridines virtually as a single isomer. When optically active nitriles were used in this reaction, chiral pyridines were obtained without loss of the enantiopurity. Alternatively, the azatitanacyclopentadiene prepared from an unsymmetrical acetylene reacted with an aldehyde or another nitrile to give furans or pyrroles having four different substituents again in a regioselective manner. PMID- 15898798 TI - Synthesis and characterization of highly luminescent CdSe-core CdS/Zn0.5Cd0.5S/ZnS multishell nanocrystals. AB - We report on the preparation and structural characterization of CdSe nanocrystals, which are covered by a multishell structure from CdS and ZnS. By using the newly developed successive ion layer adhesion and reaction (SILAR) technique, we could gradually change the shell composition from CdS to ZnS in the radial direction. Because of the stepwise adjustment of the lattice parameters in the radial direction, the resulting nanocrystals show a high crystallinity and are almost perfectly spherical, as was investigated by X-ray diffraction and electron microscopy. Also, due to the radial increase of the respective valence- and conduction-band offsets, the nanocrystals are well electronically passivated. This leads to a high fluorescence quantum yield of 70-85% for the amine terminated multishell particles in organic solvents and a quantum yield of up to 50% for mercapto propionic acid-covered particles in water. Finally, we present experimental results that substantiate the superior photochemical and colloidal stability of the multishell particles. PMID- 15898799 TI - Brightly phosphorescent trinuclear copper(I) complexes of pyrazolates: substituent effects on the supramolecular structure and photophysics. AB - Synthetic details, solid-state structures, and photophysical properties of a group of trimeric copper(I) complexes containing pyrazolate ligands are described. The reaction of copper(I) oxide and the fluorinated pyrazoles [3 (CF(3))Pz]H, [3-(CF(3)),5-(Me)Pz]H, and [3-(CF(3)),5-(Ph)Pz]H leads to the corresponding trinuclear copper(I) pyrazolates, {[3-(CF(3))Pz]Cu}(3), {[3 (CF(3)),5-(Me)Pz]Cu}(3), and {[3-(CF(3)),5-(Ph)Pz]Cu}(3), respectively, in high yield. The {[3,5-(i-Pr)(2)Pz]Cu}(3) compound was obtained by a reaction between [Cu(CH(3)CN)(4)][BF(4)], [3,5-(i-Pr)(2)Pz]H, and NEt(3). These compounds as well as {[3,5-(Me)(2)Pz]Cu}(3) and {[3,5-(CF(3))(2)Pz]Cu}(3) adopt trimeric structures with nine-membered Cu(3)N(6) metallacycles. There are varying degrees and types of intertrimer Cu...Cu interactions. These contacts give rise to zigzag chains in the fluorinated complexes, {[3-(CF(3))Pz]Cu}(3), {[3-(CF(3)),5-(Me)Pz]Cu}(3), {[3 (CF(3)),5-(Ph)Pz]Cu}(3), and {[3,5-(CF(3))(2)Pz]Cu}(3), whereas the nonfluorinated complexes, {[3,5-(Me)(2)Pz]Cu}(3) and {[3,5-(i-Pr)(2)Pz]Cu}(3) form dimers of trimers. Out of all the compounds examined in this study, {[3 (CF(3)),5-(Ph)Pz]Cu}(3) has the longest (3.848 Angstroms) and {[3,5 (Me)(2)Pz]Cu}(3) has the shortest (2.946 Angstroms) next-neighbor intertrimer Cu...Cu distance. The Cu...Cu separations within the trimer units do not vary significantly (typically 3.20-3.26 Angstroms). All of these trinuclear copper(I) pyrazolates show bright luminescence upon exposure to UV radiation. The luminescence bands are hugely red-shifted from the corresponding lowest-energy excitations, rather broad, and unstructured even at low temperatures, suggesting metal-centered emissions owing to intertrimer Cu...Cu interactions that are strengthened in the phosphorescent state. The {[3-(CF(3)),5-(Ph)Pz]Cu}(3) compound exhibits an additional highly structured phosphorescence with a vibronic structure corresponding to the pyrazolyl (Pz) ring. The luminescence properties of solids and solutions of the trimeric compounds in this study show fascinating trends with dramatic sensitivities to temperature, solvent, concentration, and excitation wavelengths. PMID- 15898800 TI - Discovery and high-throughput screening of heteroleptic iridium complexes for photoinduced hydrogen production. AB - The catalytic process of photoinduced hydrogen generation via the reduction of water has been investigated. The use of parallel synthetic techniques has facilitated the synthesis of a 32 member library of heteroleptic iridium complexes that was screened, using high-throughput photophysical techniques, to identify six potential photosensitizers for use in catalytic photoinduced hydrogen production. A Pd/Ni thin film hydrogen selective sensor allowed for rapid quantification of hydrogen produced via illumination of aqueous systems of the photosensitizer, tris(2,2'-dipyridyl)dichlorocobalt ([Co(bpy)(3)]Cl(2)), and triethanolamine (a sacrificial reductant) with ultra-bright light emitting diodes (LEDs). The use of an 8-well parallel photoreactor expedited the investigation of the hydrogen evolution process and facilitated mechanistic studies. All six compounds investigated produced considerably more hydrogen than commonly utilized photosensitizers and had relative quantum efficiencies of hydrogen production up to 37 times greater than that of Ru(bpy)(3)(2+). PMID- 15898801 TI - Quantum effects on hydrogen isotope adsorption on single-wall carbon nanohorns. AB - H(2) and D(2) adsorption on single-wall carbon nanohorns (SWNHs) have been measured at 77 K, and the experimental data were compared with grand canonical Monte Carlo simulations for adsorption of these hydrogen isotopes on a model SWNH. Quantum effects were included in the simulations through the Feynman-Hibbs effective potential. The simulation predictions show good agreement with the experimental results and suggest that the hydrogen isotope adsorption at 77 K can be successfully explained with the use of the effective potential. According to the simulations, the hydrogen isotopes are preferentially adsorbed in the cone part of the SWNH with a strong potential field, and quantum effects cause the density of adsorbed H(2) inside the SWNH to be 8-26% smaller than that of D(2). The difference between H(2) and D(2) adsorption increases as pressure decreases because the quantum spreading of H(2), which is wider than that of D(2), is fairly effective at the narrow conical part of the SWNH model. These facts indicate that quantum effects on hydrogen adsorption depend on pore structures and are very important even at 77 K. PMID- 15898802 TI - NMR detection of single-walled carbon nanotubes in solution. AB - The detection of nanotube carbons in solution by (13)C NMR is reported. The highly soluble sample was from the functionalization of (13)C-enriched single walled carbon nanotubes (SWNTs) with diamine-terminated oligomeric poly(ethylene glycol) (PEG(1500N)). The ferromagnetic impurities due to the residual metal catalysts were removed from the sample via repeated magnetic separation. The nanotube carbon signals are broad but partially resolved into two overlapping peaks, which are tentatively assigned to nanotube carbons on semiconducting (upfield) and metallic (downfield) SWNTs. The solid-state NMR signals of the same sample are similarly resolved. Mechanistic and practical implications of the results are discussed. PMID- 15898803 TI - Synthesis of a new class of compounds containing a Ln-O-Al arrangement and their reactions and catalytic properties. AB - Synthesis of a new class of compounds containing a Ln-O-Al moiety has been accomplished by the reaction of LAlOH(Me) (L = HC(CMeNAr)(2), Ar = 2,6 iPr(2)C(6)H(3)) with a series of Cp(3)Ln compounds. The terminal Al-OH group shows selective reactivity, and the complexes Cp(2)Ln(THF)-O-AlL(Me) (Ln = Yb, 1; Er, 2; Dy, 3), Cp(2)Yb-O-AlL(Me) (4), and Cp(3)Ln(mu-OH)AlL(Me) (Ln = Er, 5; Dy, 6; Sm, 7) were obtained. This allows further insight into the proton exchange process, and two different mechanisms, intermolecular and intramolecular elimination of CpH, are proposed under different conditions. Complexes 1-4, 6, and 7 have been characterized by X-ray structural analyses which reveals a Ln-O Al or Ln(mu-OH)Al core in these complexes. The obtuse Ln-O-Al angles fall in the range 151.9-169.8 degrees . The reaction of 1 or 4 with Me(3)SnF in toluene under refluxing conditions unexpectedly yielded the compounds [Cp(2)Yb(mu-OSnMe(3))](2) (8) and LAl(Me)F (9). Reactions of LAlOH(Me) with the mono- and dicyclopentadienyl complexes LYbCp(Cl) (10) and LYbCp(2) (11) supported by the bulky beta-diketiminate ligand were unsuccessful. However, the reaction of LAl(OH)Me with LYbN(SiMe(3))(2)Cl (12) containing a labile Yb-N bond leads to the formation of LYbCl-O-AlL(Me) (13) under elimination of HN(SiMe(3))(2). Furthermore, complexes 1, 3, 4, and 6 exhibit good catalytic activity for the polymerization of epsilon-caprolactone. PMID- 15898804 TI - Short- and long-range order in the positive electrode material, Li(NiMn)0.5O2: a joint X-ray and neutron diffraction, pair distribution function analysis and NMR study. AB - The local environments and short-range ordering of LiNi(0.5)Mn(0.5)O(2), a potential Li-ion battery positive electrode material, were investigated by using a combination of X-ray and neutron diffraction and isotopic substitution (NDIS) techniques, (6)Li Magic Angle Spinning (MAS) NMR spectroscopy, and for the first time, X-ray and neutron Pair Distribution Function (PDF) analysis, associated with Reverse Monte Carlo (RMC) calculations. Three samples were studied: (6)Li(NiMn)(0.5)O(2), (7)Li(NiMn)(0.5)O(2), and (7)Li(NiMn)(0.5)O(2) enriched with (62)Ni (denoted as (7)Li(ZERO)Ni(0.5)Mn(0.5)O(2)), so that the resulting scattering length of Ni atoms is null. LiNi(0.5)Mn(0.5)O(2) adopts the LiCoO(2) structure (space group Rm) and comprises separate lithium layers, transition metal layers (Ni, Mn), and oxygen layers. NMR experiments and Rietveld refinements show that there is approximately 10% of Ni/Li site exchange between the Li and transition metal layers. PDF analysis of the neutron data revealed considerable local distortions in the layers that were not captured in the Rietveld refinements performed using the Bragg diffraction data and the LiCoO(2) structure, resulting in different M-O bond lengths of 1.93 and 2.07 Angstroms for Mn-O and Ni/Li-O, respectively. Large clusters of 2400-3456 atoms were built to investigate cation ordering. The RMC method was then used to improve the fit between the calculated model and experimental PDF data. Both NMR and RMC results were consistent with a nonrandom distribution of Ni, Mn, and Li cations in the transition metal layers; both the Ni and Li atoms are, on average, close to more Mn ions than predicted based on a random distribution of these ions in the transition metal layers. Constraints from both experimental methods showed the presence of short-range order in the transition metal layers comprising LiMn(6) and LiMn(5)Ni clusters combined with Ni and Mn contacts resembling those found in the so-called "flower structure" or structures derived from ordered honeycomb arrays. PMID- 15898805 TI - Functional one-dimensional lipid bilayers on carbon nanotube templates. AB - Use of biological machines and environments in novel bioinorganic nanostructures is critical for development of new types of biosensors, bio-NEMS devices, and functional materials. Lipid bilayers that mimic a cell membrane have already played an important role in such applications. We present supported lipid bilayers that spontaneously assemble in a continuous nanoshell around a template of a carbon nanotube wrapped with hydrophilic polymer cushion layers. We demonstrate that such 1-D lipid membranes are fluid and can heal defects, even over repeated damage-recovery cycles. A simple diffusion model can describe mobility of lipid molecules in these 1-D nanoshells. These structures could lead to the development of new classes of biosensors and bioelectronic devices. PMID- 15898806 TI - Solid state NMR method for the determination of 3D zeolite framework/sorbate structures: 1H/29Si CP MAS NMR study of the high-loaded form of p-xylene in ZSM-5 and determination of the unknown structure of the low-loaded form. AB - A general protocol is described for structure determinations of organic sorbate zeolite complexes based on the selective, through-space, distance-dependent transfer of magnetization from protons in selectively deuterated organics to framework silicon nuclei. The method was developed using the known structure of the high-loaded ZSM-5/p-xylene complex containing p-xylene-d(6) or p-xylene-d(4). It was then applied to determine the unknown structure of the low-loaded ZSM-5/p xylene complex using NMR alone. For the high-loaded complex improved data were obtained below 273 K, where slow motions and exchange processes of the p-xylene are eliminated. The general approach was validated by the exact agreement of the experimental (1)H-(29)Si CPMAS spectra obtained at a specific contact time and the complete 24-line spectra simulated using 1/T(CP) vs M(2) correlations from only the six clearly resolved resonances. For the low-loaded complex the (29)Si resonances were assigned at 267 K, and variable contact time CP experiments were carried out between 243 and 173 K using the same specifically deuterated p xylenes. All possible locations and orientations of the p-xylene guests were sampled, and those solutions that gave acceptable linear 1/T(CP) vs M(2) correlations were selected. The optimum p-xylene location in this temperature range was determined to be in the channel intersection with the long molecular axis parallel to [0,1,0] (ring center fractional coordinates {-0.009, 0.250, 0.541}) with the ring plane oriented at an angle of 30 +/- 3 degrees about the crystallographic b axis. A subsequent single-crystal X-ray study confirmed this predicted structure. PMID- 15898807 TI - What is required to stabilize Al3+? A gas-phase perspective. AB - With a combination of experiment and theory (ab initio and DFT), we demonstrate that the Al(3+) cation can be stabilized in the gas phase using ligands, which have the ability to act as powerful sigma electron donors and electron acceptors. The latter property, which implies that electron density from the aluminum cation moves into ligand antibonding orbitals, has not previously been considered significant when accounting for the behavior of Al(3+). Of the three ligands identified as falling into the above category, acetonitrile appears to form the most stable complexes in the gas phase, which is in accord with the long established fact that solid-state complexes with Al(3+) are readily isolated. From the results, it is suggested that chain or ring compounds containing the -C triple bond N group might act as successful sequestering agents for Al(3+) from aqueous solutions. PMID- 15898808 TI - The molecular basis of resilience to the effect of the Lys103Asn mutation in non nucleoside HIV-1 reverse transcriptase inhibitors studied by targeted molecular dynamics simulations. AB - A series of targeted molecular dynamics simulations have been carried out in an attempt to assess the effect that the common Lys103Asn mutation in HIV-1 reverse transcriptase (RT) has on the binding of three representative non-nucleoside RT inhibitors (NNRTI), nevirapine, efavirenz, and etravirine. We have shown previously that, in the absence of an incoming inhibitor, creation of the NNRTI binding pocket is hampered due to the existence of a hydrogen bond between the side chains of Asn103 and Tyr188 for which no equivalent exists in the wild-type enzyme. As an extension of this work, we now apply the same methodology to drive the enzyme's conformation from the unbound state to the drug-bound state in the presence of the NNRTI. The location of each drug outside the binding pocket was determined by an automated docking program, and steering into the binding pocket followed a route that is likely to represent the actual entrance pathway. The additional hurdle to inhibitor entry imposed by the extra Asn103-Tyr188 hydrogen bond is seen to affect each NNRTI differently, with the ability to disrupt this interaction increasing in the order etravirine >> efavirenz > or = nevirapine, in good accord with the experimental findings. This coherent picture strongly suggests that attempts to overcome resistance through structure-based drug design may be considerably more successful if dynamic structural aspects of the type studied here are considered, particularly in cases where binding energy-based structure-activity relationship methods are unable to provide the required information. PMID- 15898809 TI - Electrochemical reduction of NO by hemin adsorbed at pyrolitic graphite. AB - The mechanism of the electrochemical reduction of nitric oxide (NO) by hemin adsorbed at pyrolitic graphite was investigated. The selectivity of NO reduction was probed by combining the rotating ring disk electrode (RRDE) technique with a newly developed technique called on-line electrochemical mass spectroscopy (OLEMS). These techniques show that NO reduction by adsorbed heme groups results in production of hydroxylamine (NH(2)OH) with almost 100% selectivity at low potentials. Small amounts of nitrous oxide (N(2)O) were only observed at higher potentials. The rate-determining step in NO reduction most likely consists of an electrochemical equilibrium involving a proton transfer, as can be derived from the Tafel slope value of 62 mV/dec and the pH dependence of -42 mV/pH. The almost 100% selectivity toward NH(2)OH distinguishes this system both from NO reduction on bare metal electrodes, which often yields NH(3), and from biological NO reduction in cytochrome P450nor, which yields N(2)O exclusively. PMID- 15898810 TI - Studies of organically functionalized mesoporous silicas using heteronuclear solid-state correlation NMR spectroscopy under fast magic angle spinning. AB - Highly resolved solid-state HETCOR NMR spectra between protons and low gamma nuclei ((13)C and (29)Si) can be suitably obtained on surfaces using a "brute force" (1)H-(1)H decoupling by MAS at rates > or =40 kHz. Despite a small rotor volume (<10 microL), a (1)H-(13)C HETCOR spectrum of allyl groups (AL, -CH(2) CH=CH(2)) covalently anchored to the surface of MCM-41 silica was acquired without using isotope enrichment. The advantages of using fast MAS in such studies include easy setup, robustness, and the opportunity of using low RF power for decoupling. In the case of the (1)H-(29)Si HETCOR experiment, the sensitivity can be dramatically increased, in some samples by more than 1 order of magnitude, through implementing into the pulse sequence a Carr-Purcell-Meiboom-Gill train of pi pulses at the (29)Si spin frequency. The use of low-power heteronuclear decoupling is essential in the (1)H-(29)Si CPMG-HETCOR experiment, due to unusually long acquisition periods. These methods provided detailed structural characterization of the surface of AL-MCM mesoporous silica. PMID- 15898811 TI - Supramolecular layer-by-layer assembly: alternating adsorptions of guest- and host-functionalized molecules and particles using multivalent supramolecular interactions. AB - The stepwise construction of a novel kind of self-assembled organic/inorganic multilayers based on multivalent supramolecular interactions between guest functionalized dendrimers and host-modified gold nanoparticles has been developed, yielding supramolecular layer-by-layer assembly. The deposition process was monitored by surface plasmon resonance spectroscopy. Further characterization of the multilayer films was performed by means of UV/vis absorption spectroscopy, which showed a linear increase in absorption with the number of bilayers. The growth of the gold nanoparticle plasmon absorption band corresponded to approximately a dense monolayer of gold nanoparticles per bilayer. Ellipsometry and atomic force microscopy (AFM) scratching experiments were used to measure the development of the film thickness with the number of bilayers, confirming linear growth and a thickness increase of approximately 2 nm/bilayer. PMID- 15898812 TI - MCM-48-like large mesoporous silicas with tailored pore structure: facile synthesis domain in a ternary triblock copolymer-butanol-water system. AB - Assembly of mesostructured silica using Pluronic P123 triblock copolymer (EO(20) PO(70)-EO(20)) and n-butanol mixture is a facile synthesis route to the MCM-48 like ordered large mesoporous silicas with the cubic Iad mesostructure. The cubic phase domain is remarkably extended by controlling the amounts of butanol and silica source correspondingly. The extended phase domain allows synthesis of the mesoporous silicas with various structural characteristics. Characterization by powder X-ray diffraction, nitrogen physisorption, scanning electron microscopy, and transmission electron microscopy reveals that the cubic Iad materials possess high specific surface areas, high pore volumes, and readily tunable pore diameters in narrow distribution of sizes ranging from 4 to 12 nm. Moreover, generation of complementary pores between the two chiral channels in the gyroid Iad structure can be controlled systematically depending on synthesis conditions. Carbon replicas, using sucrose as the carbon precursor, are obtained with either the same Iad structure or I4(1)/a (or lower symmetry), depending on the controlled synthesis conditions for silica. Thus, the present discovery of the extended phase domain leads to facile synthesis of the cubic Iad silica with precise structure control, offering vast prospects for future applications of large-pore silica materials with three-dimensional pore interconnectivity. PMID- 15898813 TI - Synthesis, optical properties, and growth mechanism of blue-emitting CdSe nanorods. AB - Blue-emitting, cubic phase CdSe nanorods with an approximate diameter of 2.5 nm and lengths up to 12 nm have been synthesized at low temperature (100 degrees C) in a single surfactant using a single-source molecular precursor. Transmission electron microscopy and dynamic light scattering measurements indicate that the nanorods are formed from self-assembly of isotropic nanoclusters. Anisotropic growth in a single surfactant appears to be favored when growth occurs below the thermal decomposition temperature of the single-source precursor. PMID- 15898814 TI - Modeling rate-controlling solvent effects. The pericyclic meisenheimer rearrangement of N-propargylmorpholine N-oxide. AB - The activation parameters of the pericyclic Meisenheimer rearrangement and a competitive rearrangement of N-propargylmorpholine N-oxide were determined by experimental and computational methods. A number of aprotic and protic solvents of different polarities and hydrogen bond-forming abilities and the roles of electron-pair acceptor additives were investigated. The reaction kinetics were followed by means of NMR. In protic solvents, isotope-labeling experiments revealed a novel inverse secondary kinetic isotope effect (k(H)/k(D) about 0.8) for the rate-determining cyclization step, probably occurring because of a C(sp) -> C(sp(2)) change in hybridization at the reaction center. In molecular computations at the B3LYP/6-31++G(d,p) level of theory, implicit, explicit, and joint explicit-implicit solvent models were used. The explicit-implicit model and molecular dynamic simulations gave the most accurate results. The components of the rate-controlling solvent effect are discussed, and general equations are proposed for accurate prediction of the solvent-dependent activation parameters. PMID- 15898815 TI - Mechanisms and applications of plasmon-induced charge separation at TiO2 films loaded with gold nanoparticles. AB - Plasmon-induced photoelectrochemistry in the visible region was studied at gold nanoparticle-nanoporous TiO(2) composites (Au-TiO(2)) prepared by photocatalytic deposition of gold in a porous TiO(2) film. Photoaction spectra for both the open circuit potential and short-circuit current were in good agreement with the absorption spectrum of the gold nanoparticles in the TiO(2) film. The gold nanoparticles are photoexcited due to plasmon resonance, and charge separation is accomplished by the transfer of photoexcited electrons from the gold particle to the TiO(2) conduction band and the simultaneous transfer of compensative electrons from a donor in the solution to the gold particle. Besides its low-cost and facile preparation, a photovoltaic cell with the optimized electron mediator (Fe(2+/3+)) exhibits an optimum incident photon to current conversion efficiency (IPCE) of 26%. The Au-TiO(2) can photocatalytically oxidize ethanol and methanol at the expense of oxygen reduction under visible light; it is potentially applicable to a new class of photocatalysts and photovoltaic fuel cells. PMID- 15898816 TI - Electrochemistry of unfolded cytochrome c in neutral and acidic urea solutions. AB - The present investigation reports the first experimental measurements of the reorganization energy of unfolded metalloprotein in urea solution. Horse heart cytochrome c (cyt c) has been found to undergo reversible one-electron transfer reactions at pH 2 in the presence of 9 M urea. In contrast, the protein is electrochemically inactive at pH 2 under low-ionic strength conditions in the absence of urea. Urea is shown to induce ligation changes at the heme iron and lead to practically complete loss of the alpha-helical content of the protein. Despite being unfolded, the electron-transfer (ET) kinetics of cyt c on a 2 mercaptoethanol-modified Ag(111) electrode remain unusually fast and diffusion controlled. Acid titration of ferric cyt c in 9 M urea down to pH 2 is accompanied by protonation of one of the axial ligands, water binding to the heme iron (pK(a) = 5.2), and a sudden protein collapse (pH < 4). The formal redox potential of the urea-unfolded six-coordinate His18-Fe(III)-H(2)O/five-coordinate His18-Fe(II) couple at pH 2 is estimated to be -0.083 V vs NHE, about 130 mV more positive than seen for bis-His-ligated urea-denatured cyt c at pH 7. The unusually fast ET kinetics are assigned to low reorganization energy of acid/urea unfolded cyt c at pH 2 (0.41 +/- 0.01 eV), which is actually lower than that of the native cyt c at pH 7 (0.6 +/- 0.02 eV), but closer to that of native bis-His ligated cyt b(5) (0.44 +/- 0.02 eV). The roles of electronic coupling and heme flattening on the rate of heterogeneous ET reactions are discussed. PMID- 15898817 TI - Ferrocenylundecanethiol self-assembled monolayer charging correlates with negative differential resistance measured by conducting probe atomic force microscopy. AB - Electrical and mechanical properties of metal-molecule-metal junctions formed between Au-supported self-assembled monolayers (SAMs) of electroactive 11 ferrocenylundecanethiol (FcC(11)SH) and a Pt-coated atomic force microscope (AFM) tip have been measured using a conducting probe (CP) AFM in insulating alkane solution. Simultaneous and independent measurements of currents and bias dependent adhesion forces under different applied tip biases between the conductive AFM probe and the FcC(11)SH SAMs revealed reversible peak-shaped current-voltage (I-V) characteristics and correlated maxima in the potential dependent adhesion force. Trapped positive charges in the molecular junction correlate with high conduction in a feature showing negative differential resistance. Similar measurements on an electropassive 1-octanethiol SAM did not show any peaks in either adhesion force or I-V curves. A mechanism involving two step resonant hole transfer through the occupied molecular orbitals (MOs) of ferrocene end groups via sequential oxidation and subsequent reduction, where a hole is trapped by the phonon relaxation, is proposed to explain the observed current-force correlation. These results suggest a new approach to probe charge transfer involving electroactive groups on the nanoscale by measuring the adhesion forces as a function of applied bias in an electrolyte-free environment. PMID- 15898818 TI - Inhaled insulins: their potential in the treatment of diabetes mellitus. AB - The inhalation of insulin was conceptualized by the mid-1920s, but the first successful testing of inhaled insulin occurred in the mid-1990s. The lung has proven to be an organ well capable of absorbing insulin in a reproducible and dose-dependent manner. At present, two concepts of pulmonary insulin delivery at relatively advanced stages of development have been investigated in several published studies. The first involves the Exubera device, a system consisting of a formulation of insulin in a dry and amorphous powder, which is then packaged into blisters. A special delivery system generates a pulse of compressed air, which causes the insulin to form a white fog in a transparent reservoir that can be inhaled by deep breathing. The second approach is the AERx insulin Diabetes Management System, which uses an aqueous formulation of insulin, delivered as an aerosol generated by a special, microprocessor-controlled, inhalation device. This device is capable of monitoring the patient's inspiratory flow and guiding the inhalation by a microelectronic feedback system. The therapeutic efficacy and safety of these inhaled insulins seem comparable to those of subcutaneous insulin regimens; however, inhaled insulins do not appear to achieve significantly better glycemic control. Several other concepts for the pulmonary delivery of insulin are also being developed. With the incidence of diabetes mellitus, especially type 2 diabetes, dramatically increasing worldwide, patients with type 2 diabetes appear to be an important target group for new modalities of insulin delivery. In this group, the onset of insulin treatment is frequently delayed due to the fear of self-injection, preventing effective glycemic control. Patient acceptance of inhaled insulins is excellent and no serious adverse effects have been observed to date. Further advantages of inhaled insulins are the more rapid onset of insulin action and a mitigation of postprandial glucose excursions. However, there are some open questions. The most important concerns the possible long-term effects of insulin inhalation on the lung, as insulin is known to have growth promoting properties. Thus far, there are no observations of the effects of inhaled insulin on lung structure and function that extend beyond 10 years. In patients with pulmonary disease, the smaller cumulative alveolar surface may cause problems in absorption, and in smokers the action of inhaled insulin has been shown to be stronger and with a faster onset. Furthermore, treatment with inhaled insulin requires larger doses of insulin compared with the subcutaneous route of insulin administration to achieve the same systemic effect, and the costs of this therapy could therefore be significantly higher than the costs of present insulin therapies. PMID- 15898819 TI - Extended-cycle oral contraception: a new option for routine use. AB - Extended use of oral contraceptive (OC) pills can successfully suppress endometrial activity and prevent menstruation for several months. Given that missed menses in women not using hormonal contraception may be of medical concern, understanding how hormonal contraceptives eliminate these concerns is important for both patient and healthcare provider acceptance. OC withdrawal bleeding is an artificial, iatrogenic event, which results from the deliberate, periodic interruption of hormonal support of the endometrium. Historically, it was important to provide periodic bleeding to reassure OC efficacy, but today it is recognized that these bleeding episodes are medically unnecessary and cause patient discomfort and out-of-pocket expenses. Decades of experience with prolonged use of OCs have been accumulated for women with specific menstrual related problems such as endometriosis, dysmenorrhea, and menstrual migraine headaches. Today there is a US FDA-approved product to routinely reduce the number of withdrawal periods. Clinical trials show that there is an initial increase in unscheduled bleeding and spotting days with extended-cycle OC use, but an absolute decrease in total days of bleeding and spotting from the first cycle of use. Over time, unscheduled bleeding and spotting decreases to rates found with the use of conventional-cycle regimens. Every woman who is interested in using OC pills should be offered the opportunity to choose how to use them, to determine if and when she will have withdrawal bleeding. PMID- 15898820 TI - Altered hypothalamic-pituitary-ovarian axis function in young female athletes: implications and recommendations for management. AB - Young women have become increasingly active in athletics during the 20th century. Those involved in sports that emphasize lean body type are at high risk for the development of menstrual dysfunction, including amenorrhea. This is mediated by an alteration in function of the hypothalamic-pituitary-ovarian (HPO) axis, with loss of normal secretion of luteinizing hormone, and subsequent lack of estrogen production. Disruption of the HPO axis appears to be dependent on the body's recognition of an energy imbalance, which may be due to a lack of compensatory caloric intake in the face of significant energy expenditure. Other pituitary hormones, such as triiodothyronine, growth hormone, and insulin-like growth factor-1 may also be affected. These metabolic changes have an impact on bone mineralization during a critical period in the development of bone mass. Recognition by physicians of the so-called 'female athlete triad', consisting of disordered eating, amenorrhea, and osteoporosis, may allow therapeutic intervention. Diagnosis of eating disorders and decreased bone mineral density can have significant impact on the health of the young athlete. Treatment is aimed at restoring normal menstrual function by increasing caloric intake to balance the increased energy demands of athletic participation. Concurrent treatment of the hypoestrogenemic state using estrogen replacement is controversial, but may aid in alleviating further loss of bone mass. PMID- 15898821 TI - Urofollitropin and ovulation induction. AB - Anovulation is a common cause of female infertility. Treatment for women with anovulation is aimed at induction of ovulation. Ovulation induction with follicle stimulating hormone (FSH) is indicated in women with WHO type II anovulation in whom treatment with clomifene citrate (clomifene) has failed. The majority of these women have polycystic ovary syndrome. The major disadvantages of ovulation induction with FSH are the risk of ovarian hyperstimulation syndrome and the risk of higher order multiple pregnancies. To reduce the rate of complications due to multiple follicular development, FSH should be administered using a chronic low dose protocol with small dose increments. In women with WHO type I anovulation, an exogenous supply of luteinizing hormone (LH) is required to achieve an adequate follicular response to FSH treatment. Thus, ovulation induction with FSH is not the treatment of choice in these women. FSH is a hormone that stimulates follicle growth and oocyte maturation. Endogenous FSH is produced by the pituitary gland and exists as a family of isohormones exhibiting distinct oligosaccharide structures. FSH for exogenous administration is derived from urine or is produced as recombinant FSH. The commercially available FSH products all contain different mixtures of FSH isoforms. To determine the effectiveness of urofollitropin (urinary-derived FSH), a comparison with the other available gonadotropins was made (i.e. recombinant FSH and human menopausal gonadotropin). Urofollitropin and recombinant FSH appear to be equally effective and well tolerated for ovulation induction. Human menopausal gonadotropin is comparably effective to urofollitropin in terms of pregnancy outcomes. It remains unclear whether human menopausal gonadotropins have a higher risk of overstimulation and ovarian hyperstimulation syndrome compared to urofollitropin in women with polycystic ovary syndrome. In practice, recombinant products are more convenient to use but are also more expensive. Therefore, if availability is not an issue but costs are, there is still a place for the use of urofollitropins for ovulation induction. PMID- 15898822 TI - Impact of compliance with oral antihyperglycemic agents on health outcomes in type 2 diabetes mellitus: a focus on frequency of administration. AB - Compliance with treatment is crucial to the optimal management of any chronic disease. Non-compliance with antihyperglycemic treatment is clearly a significant issue for patients with type 2 diabetes mellitus as it decreases the efficacy of the treatment and increases the risk of developing microvascular and macrovascular complications, therefore increasing the human and economic costs of this disease. The effect of low compliance on metabolic control has been shown to represent an increase of up to 1.4% in glycosylated hemoglobin. Achieving optimal compliance is therefore a therapeutic objective of prime importance. Many factors have been cited as contributing to poor compliance. Some of these, such as age, severe complications and disabilities, and social, educational, and financial difficulties, affect compliance with treatment in quite a significant manner, but are not modifiable by the healthcare provider. Other factors, such as the number of tablets per dose and polymedication, are modifiable but do not appear to be of major importance, whereas the frequency of administration is both an important and a modifiable factor affecting compliance with treatment. One strategy for optimization of compliance involves treatment of type 2 diabetes using oral antihyperglycemic agents with once-daily formulations. Recent data indicate that reducing the daily administration frequency of oral antihyperglycemic agents improves compliance with treatment and consequently metabolic control. Therefore, optimization of treatment through a reduction in the frequency of antihyperglycemic administration could be a valuable weapon in the battle to improve health outcomes and reduce the burden of type 2 diabetes. PMID- 15898823 TI - Black cohosh (Actaea/Cimicifuga racemosa): review of the clinical data for safety and efficacy in menopausal symptoms. AB - Since the publication of the results of the Women's Health Initiative that described the risks of hormone replacement therapy, many women are actively seeking alternative treatments for menopausal symptoms. Black cohosh (Actaea racemosa, syn. Cimicifuga racemosa) is one such alternative that has been used in the US for over 100 years. To date only two cimicifuga extracts have been tested clinically, and the current recommended dosage is 40-80 mg/day. Review of the published clinical data suggests that cimicifuga may be useful for the treatment of menopausal symptoms, such as hot flashes, profuse sweating, insomnia, and anxiety. However, the methodology used in most of the trials is poor and further clinical assessment of cimicifuga is needed. In terms of safety, transient adverse events such as nausea, vomiting, headaches, dizziness, mastalgia, and weight gain have been observed in clinical trials. A few cases of hepatotoxicity have been reported, but a direct association with the ingestion of cimicifuga has not been demonstrated. The most recent data suggest that cimicifuga is not estrogenic. PMID- 15898824 TI - Spotlight on paricalcitol in secondary hyperparathyroidism. AB - Paricalcitol (Zemplar) is a synthetic vitamin D2 analog that inhibits the secretion of parathyroid hormone (PTH) through binding to the vitamin D receptor. It is approved in the US and in most European nations for intravenous use in the prevention and treatment of secondary hyperparathyroidism associated with chronic renal failure in adult, and in the US pediatric, patients. Paricalcitol effectively reduced elevated serum PTH levels and was generally well tolerated in children and adults with secondary hyperparathyroidism associated with chronic renal failure. In well designed clinical trials, paricalcitol was as effective as calcitriol and as well tolerated in terms of the incidence of prolonged hypercalcemia and/or elevated calcium-phosphorus product (Ca x P). Thus, paricalcitol is a useful option for the management of secondary hyperparathyroidism in adults and children with chronic renal failure. PMID- 15898827 TI - The role of insulin and glucose (hyperinsulinaemia/euglycaemia) therapy in acute calcium channel antagonist and beta-blocker poisoning. AB - The inotropic effect of insulin has been long established. High-dose (0.5-1 IU/kg/hour) insulin, in combination with a glucose infusion to maintain euglycaemia (hyperinsulinaemia/euglycaemia therapy), has been proposed as a treatment for calcium channel antagonist (CCA) and beta-adrenoceptor antagonist (beta-blocker) poisonings. However, the basis for its beneficial effect is poorly understood.CCAs inhibit insulin secretion, resulting in hyperglycaemia and alteration of myocardial fatty acid oxidation. Similarly, blockade of beta(2) adrenoceptors in beta-blocker poisoning results in impaired lipolysis, glycogenolysis and insulin release. Insulin administration switches cell metabolism from fatty acids to carbohydrates and restores calcium fluxes, resulting in improvement in cardiac contractility. Experimental studies in verapamil poisoning have shown that high-dose insulin significantly improved survival compared with calcium salts, epinephrine or glucagon. In several life threatening poisonings in humans, the administration of high-dose insulin produced cardiovascular stabilisation, decreased the catecholamine vasopressor infusion rate and improved the survival rate. In a canine model of propranolol intoxication, high-dose insulin provided a sustained increase in systemic blood pressure, cardiac performance and survival rate compared with glucagon or epinephrine. In contrast, insulin had no effect on heart rate and electrical conduction in the myocardium. In another study, high-dose insulin reversed the negative inotropic effect of propranolol to 80% of control function and normalised heart rate. High-dose insulin produced a significant decrease in the left ventricular end-diastolic pressure and a significant increase in the stroke volume and cardiac output. The vasodilator effect was explained by an enhanced cardiac output leading to withdrawal of compensatory vasoconstriction. No clinical studies have yet been performed. Although not effective in all cases, we recommend hyperinsulinaemia/euglycaemia therapy in patients with severe CCA poisoning who present with hypotension and respond poorly to fluid, calcium salts, glucagon and catecholamine infusion. However, careful monitoring of blood glucose and serum potassium concentrations is required to avoid serious adverse effects. More clinical data are needed before this therapy can be recommended in beta-blocker poisoning. There is a need for large prospective clinical trials to confirm safety and efficacy of hyperinsulinaemia/euglycaemia therapy in both CCA and beta-blocker poisoning. PMID- 15898828 TI - Pharmacology, pathophysiology and management of calcium channel blocker and beta blocker toxicity. AB - Calcium channel blockers (CCB) and beta-blockers (BB) account for approximately 40% of cardiovascular drug exposures reported to the American Association of Poison Centers. However, these drugs represent >65% of deaths from cardiovascular medications. Yet, caring for patients poisoned with these medications can be extremely difficult. Severely poisoned patients may have profound bradycardia and hypotension that is refractory to standard medications used for circulatory support.Calcium plays a pivotal role in cardiovascular function. The flow of calcium across cell membranes is necessary for cardiac automaticity, conduction and contraction, as well as maintenance of vascular tone. Through differing mechanisms, CCB and BB interfere with calcium fluxes across cell membranes. CCB directly block calcium flow through L-type calcium channels found in the heart, vasculature and pancreas, whereas BB decrease calcium flow by modifying the channels via second messenger systems. Interruption of calcium fluxes leads to decreased intracellular calcium producing cardiovascular dysfunction that, in the most severe situations, results in cardiovascular collapse.Although, CCB and BB have different mechanisms of action, their physiological and toxic effects are similar. However, differences exist between these drug classes and between drugs in each class. Diltiazem and especially verapamil tend to produce the most hypotension, bradycardia, conduction disturbances and deaths of the CCB. Nifedipine and other dihydropyridines are generally less lethal and tend to produce sinus tachycardia instead of bradycardia with fewer conduction disturbances.BB have a wider array of properties influencing their toxicity compared with CCB. BB possessing membrane stabilising activity are associated with the largest proportion of fatalities from BB overdose. Sotalol overdoses, in addition to bradycardia and hypotension, can cause torsade de pointes. Although BB and CCB poisoning can present in a similar fashion with hypotension and bradycardia, CCB toxicity is often associated with significant hyperglycaemia and acidosis because of complex metabolic derangements related to these medications. Despite differences, treatment of poisoning is nearly identical for BB and CCB, with some additional considerations given to specific BB. Initial management of critically ill patients consists of supporting airway, breathing and circulation. However, maintenance of adequate circulation in poisoned patients often requires a multitude of simultaneous therapies including intravenous fluids, vasopressors, calcium, glucagon, phosphodiesterase inhibitors, high-dose insulin, a relatively new therapy, and mechanical devices. This article provides a detailed review of the pharmacology, pathophysiology, clinical presentation and treatment strategies for CCB and BB overdoses. PMID- 15898829 TI - Herbal interactions involving cytochrome p450 enzymes: a mini review. AB - The metabolism of a drug can be altered by another drug or foreign chemical, and such interactions can often be clinically significant. Cytochrome P450 (CYP) enzymes, a superfamily of enzymes found mainly in the liver, are involved in the metabolism of a plethora of xenobiotics and have been shown to be involved in numerous interactions between drugs and food, herbs and other drugs. The observed induction and inhibition of CYP enzymes by natural products in the presence of a prescribed drug has (among other reasons) led to the general acceptance that natural therapies can have adverse effects, contrary to the popular beliefs in countries where there is an active practice of ethnomedicine. Herbal medicines such as St. John's wort, garlic, piperine, ginseng, and gingko, which are freely available over the counter, have given rise to serious clinical interactions when co-administered with prescription medicines. Such adversities have spurred various pre-clinical and in vitro investigations on a series of other herbal remedies, with their clinical relevance remaining to be established. Although the presence of numerous active ingredients in herbal medicines, foods and dietary supplements complicate experimentation, the observable interactions with CYP enzymes warrant systematic studies, so that metabolism-based interactions can be predicted and avoided more readily. This article highlights the involvement of CYP enzymes in metabolism-related drug-herb interactions and the importance of gaining a mechanism-based understanding to avoid potential adverse drug reactions, in addition to outlining other contributory factors, such as pharmacogenetics and recreational habits that may compound this important health issue. PMID- 15898830 TI - Developing an analytical toxicology service: principles and guidance. AB - Many acutely poisoned patients are treated with no laboratory help other than general clinical chemistry and haematology. Emergency toxicological analyses (24 hour availability) that could influence immediate patient management such as iron, lithium and paracetamol (acetaminophen), are relatively few in number and are remarkably similar worldwide. These assays should be provided at hospitals with large accident and emergency departments. More complex, less frequently needed clinical toxicological assays that can often be offered on a less urgent basis are usually provided from regional or national centres because of the need to make best use of resources. Recommendations as to the assays that should be provided locally and at regional centres are available for the UK and US, and are generally applicable. Regional centres normally diversify into specialised therapeutic drug monitoring, urine screening for drugs of abuse, metals analysis and sometimes forensic work in order to widen the repertoire of tests available and to increase funding. Whatever the type and quantity of work undertaken and the instrumentation used, guidelines are now available delineating staff training, method validation, assay operation, quality control/quality assurance, and indeed virtually all other aspects of laboratory operation. These considerations notwithstanding, clinical interpretation of analytical results remains a difficult area and is the responsibility of the reporting laboratory, at least in the first instance. PMID- 15898831 TI - Drug-induced hypersensitivity: role in drug development. AB - Drug-induced hypersensitivity is an adverse reaction, characterised by damaging immune-mediated responses, initiated by medicine given at therapeutic doses for prevention, diagnosis or treatment. Immune-mediated drug hypersensitivity accounts for 6-10% of the adverse drug reactions, which rank between the fourth and sixth leading causes of death in the US. With <10% of all adverse drug reactions reported, the magnitude of the problem is significant, with estimates of costs >$US30 billion annually in the US (1995 value). In addition, the costs of not determining the potential of a drug to produce hypersensitivity in the pre clinical phase of drug development can be substantial. It has been estimated that the pre-clinical phase and clinical phase I, phase II and phase III costs are approximately $US6 million, $US12 million, $US12 million and $US100 million per drug, respectively (1999 values). It is important that investigational drugs with the potential to produce hypersensitivity reactions be identified as early in the development process as possible. Some adverse reactions to drugs can be avoided if drug-drug interactions are known or if there is a structure-activity relationship established. However, these methods are inadequate. Appropriate animal models of drug-induced hypersensitivity are needed, especially because hypersensitivity has been cited as the leading reason for taking drugs off the market. It is of critical importance to be able to predict hypersensitivity reactions to drugs. Most anaphylactic reactions occur in atopic individuals. Similarly, patients who have experienced other hypersensitivity reactions are more likely to have recurrent reactions. Therefore, animal models should be considered that predispose the animal to the reaction, such as the use of appropriate adjuvants and species. Using known positive controls of varying strengths, the investigator can rank the reaction against the positive controls as standards. This approach might yield greater results in a shorter period of time than using novel models. For the greatest safety, use of well understood models that have been thoroughly validated is imperative. PMID- 15898832 TI - RAG1 core and V(D)J recombination signal sequences were derived from Transib transposons. AB - The V(D)J recombination reaction in jawed vertebrates is catalyzed by the RAG1 and RAG2 proteins, which are believed to have emerged approximately 500 million years ago from transposon-encoded proteins. Yet no transposase sequence similar to RAG1 or RAG2 has been found. Here we show that the approximately 600-amino acid "core" region of RAG1 required for its catalytic activity is significantly similar to the transposase encoded by DNA transposons that belong to the Transib superfamily. This superfamily was discovered recently based on computational analysis of the fruit fly and African malaria mosquito genomes. Transib transposons also are present in the genomes of sea urchin, yellow fever mosquito, silkworm, dog hookworm, hydra, and soybean rust. We demonstrate that recombination signal sequences (RSSs) were derived from terminal inverted repeats of an ancient Transib transposon. Furthermore, the critical DDE catalytic triad of RAG1 is shared with the Transib transposase as part of conserved motifs. We also studied several divergent proteins encoded by the sea urchin and lancelet genomes that are 25%-30% identical to the RAG1 N-terminal domain and the RAG1 core. Our results provide the first direct evidence linking RAG1 and RSSs to a specific superfamily of DNA transposons and indicate that the V(D)J machinery evolved from transposons. We propose that only the RAG1 core was derived from the Transib transposase, whereas the N-terminal domain was assembled from separate proteins of unknown function that may still be active in sea urchin, lancelet, hydra, and starlet sea anemone. We also suggest that the RAG2 protein was not encoded by ancient Transib transposons but emerged in jawed vertebrates as a counterpart of RAG1 necessary for the V(D)J recombination reaction. PMID- 15898833 TI - On the number of New World founders: a population genetic portrait of the peopling of the Americas. AB - The founding of New World populations by Asian peoples is the focus of considerable archaeological and genetic research, and there persist important questions on when and how these events occurred. Genetic data offer great potential for the study of human population history, but there are significant challenges in discerning distinct demographic processes. A new method for the study of diverging populations was applied to questions on the founding and history of Amerind-speaking Native American populations. The model permits estimation of founding population sizes, changes in population size, time of population formation, and gene flow. Analyses of data from nine loci are consistent with the general portrait that has emerged from archaeological and other kinds of evidence. The estimated effective size of the founding population for the New World is fewer than 80 individuals, approximately 1% of the effective size of the estimated ancestral Asian population. By adding a splitting parameter to population divergence models it becomes possible to develop detailed portraits of human demographic history. Analyses of Asian and New World data support a model of a recent founding of the New World by a population of quite small effective size. PMID- 15898834 TI - A Delphi consensus on criteria for contraindications, assessment indicators and expected outcomes related to tibialis posterior transfer surgery. AB - A team of experts in the field of reconstructive surgery for leprosy-affected people was identified. Using the Delphi method, an exercise was undertaken to ascertain whether a consensus on essential criteria and indicators for Tibialis Posterior Transfer (TPT) could be reached among the team. This paper describes the Delphi Exercise, giving results at each stage of consensus development. The final outcome was that essential criteria, including contraindications for surgery, pre- and post- operative assessments and expected outcomes, were agreed. The criteria are presented with recommendations. PMID- 15898835 TI - Anesthesia of face uncovered by histopathology. PMID- 15898836 TI - Leprosy research declines, but most of the basic questions remain unanswered. PMID- 15898837 TI - Consensus methods: a bridge between clinical reasoning and clinical research? AB - Evidence-Based Practice does head the "hierarchy of evidence" upon which developments in clinical practice should be based. There are, however, situations where evidence is either unavailable, unclear, or results between studies are at variance. Consensus is a reliable contingency, and approaches to reaching consensus have acceptable construct validity (Nominal Group Technique, Delphi, and Consensus Development Conference). Consensus is reached when: (i) the method of investigation tightly controls communication to reduce the obscuring "noise" of divergent discussion; (ii) statistical measures of agreement or dissent screen out the bias that would otherwise be produced by the dictate of vociferous minorities or coalitions that may represent vested interests; (iii) all participants contribute equally to the product of the investigation. PMID- 15898838 TI - A potentially new treatment for tuberculosis; will a diarylquinoline work for leprosy? PMID- 15898839 TI - Some considerations on the origin of type 1 reactions in leprosy. PMID- 15898840 TI - Neuropathic pain in leprosy patients. PMID- 15898843 TI - Ulcerative cutaneous mycobacteriosis due to Mycobacterium ulcerans: report of two Mexican cases. AB - We report two patients from Central Mexico, with ulcerated cutaneous lesions containing acid-fast bacilli (AFB) and ultimately diagnosed as Mycobacterium ulcerans disease. The first patient had a long history (11 years) of disease involving multiple lesions of both upper and lower extremities. Histopathological changes included necrosis of the subcutaneous tissue with large numbers of extracellular AFB. Cultures at 32 degrees C were "positive for mycobacteria," but were not further identified. The polymerase chain reaction for M. ulcerans performed on skin bopsies was positive. The lesions improved after treatment with rifampin and isoniazid (INH) for one month, followed by ethambutol and streptomycin. The second case followed trauma to the right hand, which spread over 2 years to the right upper extremity, the back, and both legs, with a loss of digits and metacarpal bones of the right hand. The histopathological findings were similar to the first case, including presence of AFB. PCR for M. ulcerans on extracts of skin biopsies was positive. Rifampin, INH, pyrazinamide, and levofloxacin resulted in marked improvement of the ulcer; ethambutol and streptomycin were later used, also. We report these cases because they are rare (approximately 6 previous cases were reported from Mexico), and both are unusually disseminated. They are significant in alerting the medical community to M. ulcerans infection, which is still active in Mexico, and the treatment used has not been reported previously. PMID- 15898846 TI - School nurse work environment: ingredients for safe and effective practice. PMID- 15898847 TI - The United Kingdom National Healthy School Standard: a framework for strengthening the school nurse role. AB - The purpose of this review is to analyze the school nursing role within the National Healthy School Standard (NHSS) in the United Kingdom with a view toward clarifying and strengthening the role of school nurses globally. Within the National Healthy School Standard framework, school nurses serve an integral role in linking health and education partnerships to promote effective school health programs. School nurse contributions to the National Healthy School Standard, as well as barriers and supports, are discussed. Additionally, the methods school nurses implement to partner, to manage service delivery, and to work with schools are outlined. The central role of school nurses within the National Healthy School Standard framework provides a guide for school nurses in the United States to demonstrate their importance as key players in healthy schools that promote health and education. The framework deserves recognition as a foundational model to help strengthen both the school nurse role and school health programs around the world. PMID- 15898848 TI - Fetal alcohol spectrum disorder. AB - Fetal alcohol spectrum disorder (FASD) is a serious and widespread problem in this country. Positioned within the community with links to children, families, and healthcare systems, school nurses are a critical element in the prevention and treatment of those affected by fetal alcohol spectrum disorder. Although most school nurses are familiar with fetal alcohol syndrome (FAS) and the problems it poses, they may not be familiar with the newer term, fetal alcohol spectrum disorder. This article summarizes the most recent information about FASD and recently published guidelines on diagnostic criteria for FAS. Also outlined are primary, secondary, and tertiary prevention activities school nurses can take to help assure that children, families, and communities work to prevent the problem and to treat the children affected by it. PMID- 15898849 TI - School phobia. AB - School phobia is a serious disorder affecting up to 5% of elementary and middle school children. Long-term consequences include academic failure, diminished peer relationships, parental conflict, and development of additional psychiatric disorders. Hiding behind such common physical symptoms as headaches, stomachaches, and fatigue, school phobia evades diagnosis with ease. Unraveling the problem of school phobia is challenging for the school nurse and is complicated by an overall lack of knowledge regarding the serious potential outcomes. The purpose of this article is to define school phobia, differentiate it from truancy, and highlight some interventions useful in the treatment of this increasingly common and potentially serious disorder. PMID- 15898850 TI - Bipolar disorder in school-age children. AB - This article examines the individual components of bipolar disorder in children and the behaviors that can escalate as a result of misdiagnosis and treatment. The brain/behavior relationship in bipolar disorders can be affected by genetics, developmental failure, or environmental influences, which can cause an onset of dramatic mood swings and dysfunctional behavior. School is often the site where mental health disorders are observed when comparing behaviors with other children. Assessing the emotional, academic, and health needs of a student with a bipolar disorder is a critical step in designing effective interventions and school accommodations. Without appropriate medical, psychological, pharmaceutical, and academic interventions, a child is at risk for uncontrolled mania, depression, substance abuse, or suicide. The school nurse is part of the multidisciplinary team and plays a key role in facilitating case management to potentially reverse this possible negative trajectory. Successful case management provides children with bipolar disorder the opportunity to reach their academic potential. PMID- 15898851 TI - Lessons learned: one experience with focus groups in a school setting. AB - Focus group research is an effective way for school nurses to gain knowledge about a diverse range of issues that children in schools may face. It is a means to gather information about perceptions, feelings, points of view, and manners of thinking from a group of individuals about a specific topic. This article overviews focus groups as a research methodology and describes an experience of using them to gain information about low-income caretakers' knowledge of childhood nutrition. Several important lessons were learned from this study. These included the need to be flexible during the research process, to be creative in recruitment strategies, to maintain open communication with all the involved parties, and to share enthusiasm for the study. Learning more about children and their families can assist school nurses in planning and implementing interventions that enhance health and school achievement. PMID- 15898852 TI - Recommendations for constructing school nurses' offices designed to support school health services. AB - When attempting to locate guidelines for designing school nurses' offices, one finds there are minimal resources that address this need. In an effort to solve this concern, the authors were instrumental in writing an article for The Journal of School Nursing published in April 1997. The current article is an updated perspective that defines recommendations to be utilized in new construction or renovation of school nurses' offices. It encompasses information on square footage, design, utilization of space, location, and necessary equipment. Prototypes are provided that apply to elementary, middle school, and high school settings. The article outlines the importance of continued collaboration between the school nurse and key people involved in the planning and construction process, such as the school principal, administrators, and the district engineer. PMID- 15898853 TI - New construction, renovation and remodeling: what school nurses have learned from planning new health office facilities. PMID- 15898854 TI - What are the odds? Random drug testing of students: a legal perspective. AB - Companion articles explore random drug testing programs in schools. The first article addresses random drug testing of students from a legal perspective. It describes legal issues and current case law regarding drug testing programs in schools and commonly asked questions regarding these programs. The second article addresses random drug testing of students from a practice perspective. It explores arguments for and against random drug testing programs and the role of the school nurse in policy and procedure development. PMID- 15898855 TI - What are the odds? Random drug testing of students: a practice perspective. AB - Companion articles explore random drug testing programs in schools. The first article addresses random drug testing of students from a legal perspective. It describes legal issues and current case law regarding drug testing programs in schools and commonly asked questions regarding these programs. The second article addresses random drug testing of students from a practice perspective. It explores arguments for and against random drug testing programs and the role of the school nurse in policy and procedure development. PMID- 15898857 TI - Lumps, bumps, and things that go itch in your office! PMID- 15898858 TI - Can the theory of planned behavior predict the maintenance of physical activity? AB - This study tested the ability of the theory of planned behavior to predict actual participation in physical activity and explored the development of activity habits in a 12-week longitudinal study. People enrolling in a gymnasium (N = 94) completed standard theory of planned behavior measures at baseline and follow-up; behavior was monitored objectively in the intervening period. The data were analyzed by using both standard and repeatable events survival analysis. Results showed that (a) perceived behavioral control was significantly predictive of intentions and actual behavior, (b) stable exercise habits developed in the first 5 weeks of the study, and (c) successful prior performance enhanced perceptions of behavioral control. The implications for developing theory-based interventions that promote the maintenance of health behavior are discussed. PMID- 15898859 TI - The impact of breast cancer on the lives of middle-aged women: results from the Australian longitudinal study of women's health. AB - This article investigated the impact of breast cancer (BC) in middle-aged Australian women (45-50 years). Two waves of data collected 2 years apart from a longitudinal survey of 12,177 women identified 3 groups: (a) 11,933 (98%) who reported never having had BC, (b) 181 (1.5%) who reported a diagnosis of BC at Time 1, and (c) 63 (0.5%) who reported onset of BC between Time 1 and Time 2. Repeated measures analysis of variance was used to compare the 3 groups. Women with recent onset of BC experienced significant changes across a range of functioning compared with the other 2 groups. Compared with women with no BC, women with longer established onset of BC had significantly worse health and social outcomes, but these were associated with small effect sizes. Both groups of women with BC reported less impact on mental and emotional health than on other areas of functioning. PMID- 15898860 TI - Daily mood, partner support, sexual interest, and sexual activity among adolescent women. AB - The main objective of this study was to examine day-to-day associations of coitus, sexual interest, partner emotional support, negative mood, and positive mood among adolescent women. Diaries assessed partner interactions, sexual activity, substance use, and mood. Participants were 146 adolescent women who provided 28,376 diary days. Correlates of coitus on a given day included age, increased coital frequency in previous week, coitus on the previous day, partner support, increased same-day sexual interest, and decreased same-day negative mood. The data demonstrate complex associations of sexual interest, mood, partner interactions, and sexual activity. The findings extend understanding of the sexuality of adolescent women and have implications for a variety of interventions to reduce sexually transmitted infections and unplanned pregnancies. PMID- 15898861 TI - Unfair treatment, discrimination, and ambulatory blood pressure in black and white adolescents. AB - The authors tested the hypotheses that unfair treatment and its attribution to race, physical appearance, and peer group were related to elevated ambulatory blood pressure (ABP). During 2 school days, 207 Black and White adolescents wore an ABP monitor and answered questions about mood, posture, location, and activity level at the time of the ABP assessment. At a separate session, in-clinic resting blood pressure and perceptions of unfair treatment were measured. Multilevel mixed models showed that unfair treatment and its attribution to race were not associated with ABP. However, adolescents who indicated that the primary reason for unfair treatment was their physical appearance had elevated ABP. Feeling unfairly treated because of physical appearance may impact blood pressure uniquely during the adolescent transition. PMID- 15898862 TI - Predictors of posttraumatic growth following bone marrow transplantation for cancer. AB - There is growing recognition that the experience of cancer can have a positive as well as a negative psychological impact. This longitudinal study sought to identify predictors of posttraumatic growth among cancer patients (N=72) undergoing bone marrow transplantation. Greater posttraumatic growth in the posttransplant period was related to younger age; less education; greater use of positive reinterpretation, problem solving, and seeking alternative rewards as coping strategies in the pretransplant period; more stressful appraisal of aspects of the transplant experience; and more negatively biased recall of pretransplant levels of psychological distress. Findings partially support J. A. Schaefer and R. H. Moos's (1992) model of life crises and personal growth and also suggest that temporal self-comparisons contribute to the experience of posttraumatic growth. PMID- 15898863 TI - Child behavior and parent management strategies at mealtimes in families with a school-age child with cystic fibrosis. AB - Parent and child mealtime behaviors in school-age children with cystic fibrosis (CF; n = 28) and children without CF (n = 28) were examined during dinner meals by using multivariate analysis of variance. Parents of children with CF were found to differ from comparison parents in the frequency and rate of child management strategies. No differences were found in child behaviors. As the meal progressed, children displayed an increase in behaviors incompatible with eating, and parents increased behaviors to encourage eating. Slow eaters (> 20 min) with CF consumed more calories at the dinner meal than fast eaters (< 20 min) but did not achieve a higher daily caloric intake. Interventions targeting improvement of parent- child interactions during the mealtime are needed to achieve optimal dietary intake. PMID- 15898864 TI - Self-efficacy and compliance with benzodiazepine taper in older adults with chronic insomnia. AB - Better understanding of compliance with BZD taper is warranted. Compliance with a taper program and perceived self-efficacy (SE) in being able to comply with hypnotic reduction goals was monitored weekly in 52 older adults (mean age: 63.0 years) with chronic insomnia (average duration: 21.9 years) who underwent a 10 week physician-supervised medication tapering. One group received cognitive- behavior therapy for insomnia during discontinuation, whereas the other did not. Compliant patients showed higher SE ratings at Weeks 6, 8, 9, and 10. Medication free patients at the end of the treatment also reported higher mean SE ratings at those 4 weeks. Differences remained significant when withdrawal symptoms and sleep efficiency were controlled for. These results have important clinical implications because SE may indicate key time points when patients are experiencing more difficulty during discontinuation. PMID- 15898866 TI - Loneliness, social network size, and immune response to influenza vaccination in college freshmen. AB - Antibody response to the influenza immunization was investigated in 83 1st semester healthy university freshmen. Elevated levels of loneliness throughout the semester and small social networks were independently associated with poorer antibody response to 1 component of the vaccine. Those with both high levels of loneliness and a small social network had the lowest antibody response. Loneliness was also associated with greater psychological stress and negative affect, less positive affect, poorer sleep efficiency and quality, and elevations in circulating levels of cortisol. However, only the stress data were consistent with mediation of the loneliness-antibody response relation. None of these variables were associated with social network size, and hence none were potential mediators of the relation between network size and immunization response. PMID- 15898867 TI - Temperament in childhood predicts body mass in adulthood: the Cardiovascular Risk in Young Finns Study. AB - This study examined associations of temperament at ages 6 to 12 with body-mass index (BMI) and waist circumference (WC) at ages 24 to 30 years. The participants were 619 men and women derived from the population-based Cardiovascular Risk in Young Finns Study. Temperament was operationalized as (negative) emotionality, sociability, and activity. High emotionality predicted increased BMI, independently of WC, and independently of childhood and adulthood risk factors for adult obesity. None of the temperament dimensions had any associations with WC after controlling for BMI. The findings suggest that temperamental difficulty in childhood may be a useful risk indicator for general body mass in adulthood, and the mechanisms relating temperament with body mass should be further explored. PMID- 15898865 TI - Traumatic stress, perceived global stress, and life events: prospectively predicting quality of life in breast cancer patients. AB - The authors investigated the relationship between stress at initial cancer diagnosis and treatment and subsequent quality of life (QoL). Women (n = 112) randomized to the assessment-only arm of a clinical trial were initially assessed after breast cancer diagnosis and surgery and then reassessed at 4 months (during adjuvant treatment) and 12 months (postadjuvant treatment). There were 3 types of stress measured: number of stressful life events (K. A. Matthews et al., 1997), cancer-related traumatic stress symptoms (M. J. Horowitz, N. Wilner, & W. Alvarez, 1979), and perceived global stress (S. Cohen, T. Kamarck, & R. Mermelstein, 1983). Using hierarchical multiple regressions, the authors found that stress predicted both psychological and physical QoL (J. E. Ware, K. K. Snow, & M. Kosinski, 2000) at the follow-ups (all ps < .03). These findings substantiate the relationship between initial stress and later QoL and underscore the need for timely psychological intervention. PMID- 15898868 TI - Comparative optimism about health and nonhealth events in 8- and 9-year-old children. AB - Comparative optimism has been studied extensively in adults and is a significant component of social- cognitive models about health. In contrast, little is known about comparative optimism in children or about the wider social- cognitive processes that underpin their health-related behavior. This study investigated comparative optimism for health- and nonhealth-related topics in 101 children 8 or 9 years of age, the youngest ages that have been investigated so far. Children were shown to be unrealistically optimistic for health and nonhealth events. The implications of these findings for understanding comparative optimism in children are discussed. PMID- 15898869 TI - Why are you bringing up condoms now? The effect of message content on framing effects of condom use messages. AB - According to prospect theory (A. Tversky & D. Kahneman, 1981), messages advocating a low-risk (i.e., easy, low-cost) behavior are most effective if they stress the benefits of adherence (gain framed), whereas messages advocating a risky behavior are most effective if they stress the costs of nonadherence (loss framed). Although condom use is viewed as a low-risk behavior, it may entail risky interpersonal negotiations. Study 1 (N = 167) compared ratings of condom use messages advocating relational behaviors (e.g., discussing condoms) or health behaviors (e.g., carrying condoms). As predicted, loss-framed relational messages and gain-framed health messages received higher evaluations. Study 2 (N = 225) offers a replication and evidence of issue involvement and gender as moderators. Results are discussed with reference to the design of condom use messages. PMID- 15898870 TI - Partner's adjustment to breast cancer: a critical analysis of intervention studies. AB - Partners of breast cancer patients do not have resources available for dealing with their concerns. An analysis of intervention studies with partners was conducted, spanning research published from 1966 to 2004. Although there is considerable descriptive research documenting the need for partner interventions in the context of breast cancer, only 4 studies met criteria for inclusion in this analysis. Two studies reported limited intervention efficacy, but none incorporated all characteristics of a rigorous clinical trial with adequate power to fully test the intervention. Future intervention research should incorporate randomized, controlled clinical trial designs; have adequate statistical power; clearly report eligibility criteria; delineate theoretically based, fully explicated, and consistently delivered interventions; and use outcome measures that are sensitive to empirically derived partner-adjustment issues. PMID- 15898871 TI - Counterfactual thinking and the first instinct fallacy. AB - Most people believe that they should avoid changing their answer when taking multiple-choice tests. Virtually all research on this topic, however, has suggested that this strategy is ill-founded: Most answer changes are from incorrect to correct, and people who change their answers usually improve their test scores. Why do people believe in this strategy if the data so strongly refute it? The authors argue that the belief is in part a product of counterfactual thinking. Changing an answer when one should have stuck with one's original answer leads to more "if only . . ." self-recriminations than does sticking with one's first instinct when one should have switched. As a consequence, instances of the former are more memorable than instances of the latter. This differential availability provides individuals with compelling (albeit illusory) personal evidence for the wisdom of always following their 1st instinct, with suboptimal test scores the result. PMID- 15898872 TI - Feeling and believing: the influence of emotion on trust. AB - The authors report results from 5 experiments that describe the influence of emotional states on trust. They found that incidental emotions significantly influence trust in unrelated settings. Happiness and gratitude--emotions with positive valence--increase trust, and anger--an emotion with negative valence- decreases trust. Specifically, they found that emotions characterized by other person control (anger and gratitude) and weak control appraisals (happiness) influence trust significantly more than emotions characterized by personal control (pride and guilt) or situational control (sadness). These findings suggest that emotions are more likely to be misattributed when the appraisals of the emotion are consistent with the judgment task than when the appraisals of the emotion are inconsistent with the judgment task. Emotions do not influence trust when individuals are aware of the source of their emotions or when individuals are very familiar with the trustee. PMID- 15898873 TI - Attitude importance and the accumulation of attitude-relevant knowledge in memory. AB - People who attach personal importance to an attitude are especially knowledgeable about the attitude object. This article tests an explanation for this relation: that importance causes the accumulation of knowledge by inspiring selective exposure to and selective elaboration of relevant information. Nine studies showed that (a) after watching televised debates between presidential candidates, viewers were better able to remember the statements made on policy issues on which they had more personally important attitudes; (b) importance motivated selective exposure and selective elaboration: Greater personal importance was associated with better memory for relevant information encountered under controlled laboratory conditions, and manipulations eliminating opportunities for selective exposure and selective elaboration eliminated the importance-memory accuracy relation; and (c) people do not use perceptions of their knowledge volume to infer how important an attitude is to them, but importance does cause knowledge accumulation. PMID- 15898874 TI - Different emotional reactions to different groups: a sociofunctional threat-based approach to "prejudice". AB - The authors suggest that the traditional conception of prejudice--as a general attitude or evaluation--can problematically obscure the rich texturing of emotions that people feel toward different groups. Derived from a sociofunctional approach, the authors predicted that groups believed to pose qualitatively distinct threats to in-group resources or processes would evoke qualitatively distinct and functionally relevant emotional reactions. Participants' reactions to a range of social groups provided a data set unique in the scope of emotional reactions and threat beliefs explored. As predicted, different groups elicited different profiles of emotion and threat reactions, and this diversity was often masked by general measures of prejudice and threat. Moreover, threat and emotion profiles were associated with one another in the manner predicted: Specific classes of threat were linked to specific, functionally relevant emotions, and groups similar in the threat profiles they elicited were also similar in the emotion profiles they elicited. PMID- 15898875 TI - Policewomen acting in self-defense: can psychological disengagement protect self esteem from the negative outcomes of relative deprivation? AB - The role of 2 components of psychological disengagement (discounting and devaluing) in the relation between personal relative deprivation and self-esteem was explored in 3 samples of policewomen. Path analyses conducted with the 3 samples revealed that stronger feelings of personal relative deprivation resulted in stronger discounting of work evaluations, which in turn led to devaluing the importance of police work. A negative relation between discounting and self esteem was observed in all samples. Other related outcomes of disengagement, professional withdrawal and stress, were also evaluated. PMID- 15898876 TI - Self-esteem and favoritism toward novel in-groups: the self as an evaluative base. AB - The self-as-evaluative base (SEB) hypothesis proposes that self-evaluation extends automatically via an amotivated consistency process to affect evaluation of novel in-groups. Four minimal group studies support SEB. Personal trait self esteem (PSE) predicted increased favoritism toward a novel in-group that, objectively, was equivalent to the out-group (Study 1). This association was independent of information-processing effects (Study 1), collective self-esteem, right-wing authoritarianism (RWA), and narcissism (Studies 2 and 3). A self affirmation manipulation attenuated the association between in-group favoritism and an individual difference associated with motivated social identity concerns (RWA) but did not alter the PSE effect (Study 3). Finally, the association between PSE and in-group favoritism remained positive even when the in-group was objectively less favorable than the out-group (Study 4). PMID- 15898877 TI - Having an open mind: the impact of openness to experience on interracial attitudes and impression formation. AB - This article considers how Openness to Experience may mitigate the negative stereotyping of Black people by White perceivers. As expected, White individuals who scored relatively high on Openness to Experience exhibited less prejudice according to self-report measures of explicit racial attitudes. Further, White participants who rated themselves higher on Openness to Experience formed more favorable impressions of a fictitious Black individual. Finally, after observing informal interviews of White and Black targets, White participants who were more open formed more positive impressions of Black interviewees, particularly on dimensions that correspond to negative racial stereotypes. The effect of Openness to Experience was relatively stronger for judgments of Black interviewees than for judgments of White interviewees. Taken together these findings suggest that explicit racial attitudes and impression formation may depend on the individual characteristics of the perceiver, particularly whether she or he is predisposed to consider stereotype-disconfirming information. PMID- 15898878 TI - Resilience to loss in bereaved spouses, bereaved parents, and bereaved gay men. AB - Recent research has indicated that many people faced with highly aversive events suffer only minor, transient disruptions in functioning and retain a capacity for positive affect and experiences. This article reports 2 studies that replicate and extend these findings among bereaved parents, spouses, and caregivers of a chronically ill life partner using a range of self-report and objective measures of adjustment. Resilience was evidenced in half of each bereaved sample when compared with matched, nonbereaved counterparts and 36% of the caregiver sample in a more conservative, repeated-measures ipsative comparison. Resilient individuals were not distinguished by the quality of their relationship with spouse/partner or caregiver burden but were rated more positively and as better adjusted by close friends. PMID- 15898879 TI - Gender similarities and differences in children's social behavior: finding personality in contextualized patterns of adaptation. AB - This research examined how a contextualist approach to personality can reveal social interactional patterns that are obscured by gender comparisons of overall behavior rates. For some behaviors (verbal aggression), girls and boys differed both in their responses to social events and in how often they encountered them, yet they did not differ in overall behavior rates. For other behaviors (prosocial), gender differences in overall rates were observed, yet girls and boys differed more in their social environments than in their responses to events. The results question the assumption that meaningful personality differences must be manifested in overall act trends and illustrate how gender differences in personality can be conceptualized as patterns of social adaptation that are complex and context specific. PMID- 15898880 TI - The factor structure of Greek personality adjectives. AB - Personality descriptors--3,302 adjectives--were extracted from a dictionary of the modern Greek language. Those terms with the highest frequency were administered to large samples in Greece to test the universality of the Big-Five dimensions of personality in comparison to alternative models. One- and 2-factor structures were the most stable across variable selections and subsamples and replicated such structures found in previous studies. Among models with more moderate levels of replication, recently proposed 6- and 7-lexical-factor models were approximately as well replicated as the Big Five. An emic 6-factor structure showed relative stability; these factors were labeled Negative-Valence/Honesty, Agreeableness/Positive Affect, Prowess/Heroism, Introversion/Melancholia, Even Temper, and Conscientiousness. PMID- 15898882 TI - [Spanish scientific production in diagnostic and therapeutic research area of respiratory system in international journals from 1990 to 2002]. AB - OBJECTIVE: The aim of this study were: to analyze the spanish production in diagnostic and therapeutics research area of respiratory system in international journals from 1990 to 2002. MATERIAL AND METHODS: Papers published in diagnostic and therapeutics area of respiratory system during this period of time were selected by the PUBMED system. We delimited the production with key words: respiratory system and the one that is used by the diagnostic and therapeutics research area. RESULTS: We obtain a total of 67 document published in international journals by spanish authors. The scientific production in the diagnostic and therapeutics area have stabilized in this 12 years period. The distribution of articles by the institutional affiliation and province of authors also showed a wide dispersion: Barcelona and Clinic and Provincial Hospital of Barcelona were responsible for 43.1% and 23.8% of all the production. Bronchoalveolar lavage and the use of telescopic catheter were the type of diagnostic and therapeutic procedure more productive. CONCLUSIONS: The scientific production in the diagnostic and therapeutics area of respiratory system have stabilized in this 12 years period, this evidence contrast with the high growth detected in respiratory system in general. PMID- 15898883 TI - [Drug interactions in the prescription of medical patients at hospital discharge]. AB - OBJECTIVES: To detect and evaluate the clinical significance of drugs interactions in patients discharged from hospital. MATERIAL AND METHODS: We retrospectively screened the medication for potential drug interactions of 412 patients discharged. Interactions were catalogued according to clinical importance following the Hansten and Horn's classification. RESULTS: Three hundred twenty-nine potential interactions were detected. The 39.9% of the patients had at least one potentially interacting drug combination. The 52.6% of the interactions were catalogued as Class 3, bearing in mind minimizing the risk of the interaction. We did not find any Class 1 or 2 interactions, which have potentially major severity. Oral anticoagulants and digoxin were the most frequently implicated drugs. The patient monitoring was well done in the 100% of the interactions of Oral anticoagulants with other drug, but in the interactions of digoxin with another, this control was not done adequately. One patient was rehospitalised due to high levels of digoxin, he had been discharged with two potential interactions. CONCLUSIONS: The frequently of potential drug interactions in medical patients at hospital discharged was high, but the clinical significance appear to be low. PMID- 15898884 TI - [Spondylodiskitis caused by Candida albicans. Report of two new cases]. AB - Recently, candidiasis infection is on the increase and several factors have been associated, such us immunodepression, catheters, weakening diseases, prolonged antibiotherapy, HIV infection and IDU. Spondylodiskitis due to Candida sp. is a rare entity. Two cases of spondylodiskitis due to Candida albicans were diagnosed: a 29 year old man and a 35 year old woman. Both were IDUs with a previous history of HIV infection. The most prominent clinical features in both cases were pain, fever and constitutional syndrome. We describe the clinical, diagnostic, and therapeutic features of both cases. PMID- 15898885 TI - [Pain and amyotrophy in upper members in a cirrhotic patient with Pott's disease]. AB - We report the case of a 56 year old cirrhotic woman who presented during the course of a tuberculous spondylodiscitis affecting T9-T10, a clinical picture consistent with neuralgic amyotrophy affecting the right shoulder first, and later also the left one (Parsonage-Turner syndrome). This is an uncommonly diagnosed entity of unknown etiology and pathogenesis. Magnetic resonance images (MRI) include high signal intensity in supra and infraspinatus muscles and other muscles of the shoulder girdle, compatible with muscle oedema associated with denervation. These features, combined with the ability of MRI to exclude local problems as tendinitis stresses the importance of this technique in the diagnostic evaluation of patients with neuralgic amyotrophy. PMID- 15898886 TI - [Pure autonomic failure: case and discussion]. AB - Pure autonomic failure is an a very infrequent disease, included in the group of sinucelopathies with multisystemic atrophy and Lewy disease. We present a case admitted at our service for orthostatics syncopes, and a review of the disease, its diagnosis and its treatment. PMID- 15898887 TI - [Nonalcoholic steatohepatitis, the enigma of bad progression]. AB - Still, very little is known about the precise pathogenetic mechanisms, the triggering events and in particular, the evolution and treatment of nonalcoholic steatohepatitis (NASH). It is part of the broad spectrum of nonalcoholic fatty liver diseases (NAFLDs). Mainly, it has been reported as a benign disease, associated with metabolic disorders commonly occurrence en the general population. Nevertheless, the syndrome can lead to cirrhosis, liver failure or hepatocellular carcinoma, requiring liver transplantation. We present one patient with diagnosis of NASH, who was treated initially for overweight, HTA and hyperlipaemia with incompleted response and who showed a quickly progress to cirrhosis but no cause of liver decompensated disease could be identified. Currently she is at end-stage waiting a liver transplantation. Controlled and multicentric studies with the same definition of NASH and the study end-points are needed, and will provide information about diagnosis features and novel therapies to early management of the disease. PMID- 15898888 TI - [Painful disorder and vertigo related to the ear. Frequent referral to the ENT specialist]. AB - We revise the temporomandibular dysfunction or myofacial painful syndrome, a clinical entity that shows a high prevalence at the ear, nose and throat (ENT) specialist's surgery. They are polysymptomatic patients seen at the ENT , which are sent by the Internal Medicine and family doctors. As symptoms the patients present vertiginous feeling, tinnitus and/or deafness. We discuss the concept, the ethiopathogenia, diagnosis and the treatment of this syndrome. For the ENT specialist it is a challenge to choose the most appropriate type of treatment in order to obtain a satisfying result. In spite of the great amount of issues referring to this subject, there are still controversies regarding the orientation of the patients and the type of treatment to use. PMID- 15898889 TI - [A new case of lactic acidosis caused by buformin]. PMID- 15898890 TI - [Peripheral facial diplegia as first manifestation of Guillain-Barre syndrome secondary to acute cytomegalovirus infection]. PMID- 15898891 TI - [Portal hypertension due to cavernomatosis of the portal vein]. PMID- 15898892 TI - [Atypical presentation in a new case of latent late neurosyphilis]. PMID- 15898893 TI - [Acute liver failure: prognostic criteria and transplantation]. PMID- 15898894 TI - [Rupture of branch pulmonary artery during haemodynamic study]. PMID- 15898895 TI - [Partial small bowel obstruction due to von Recklinghausen's disease]. PMID- 15898896 TI - [Haemobilia as complication of transjugular liver biopsy]. PMID- 15898897 TI - [Late respiratory insufficiency by restriction after inguinal hernioplasty]. PMID- 15898898 TI - Preliminary assessment of postoperative adhesion formation after laser-assisted mesh fixation to the peritoneal surface. AB - OBJECTIVE: This study evaluated the incidence and persistence of adhesions following intraperitoneal onlay mesh fixation with tissue soldering in an experimental model. METHODS: Anesthetized New Zealand white rabbits (n = 21), weighing 2.8-3.2 kg, underwent laparotomy. Controls (group 1 [n = 3]) had 2 x 2 cm Mersilene (Ethicon, Somerville, New Jersey) polyester mesh segments fixed to the peritoneum with staples (USSC, Norwalk, Connecticut). Group 2 (n = 7) rabbits had Mersilene mesh affixed by melting 55% collagen solder using a prototype laser (1.43 micro, 2.5 W CW, 4 mm spot size, 60 degrees C set temperature) over mesh. Group 3 (n = 6) rabbits had Vicryl (Ethicon, Somerville, New Jersey) polyglactin mesh embedded in 60% collagen solder placed onto the peritoneum and fixed with identical laser parameters as group 2. Group 4 (n = 5) rabbits had 55% collagen solder placed and Mersilene pressed into it after melting. Four segments were placed in each experimental animal. Animals were euthanized at 2, 4, or 6 weeks. Adhesions were graded (0 = none; I = filmy adhesions; II = omental; III = bowel adhesions gently lysed; IV = dense adhesions requiring sharp dissection). RESULTS: Grade III adhesions were observed in both control and group 4 animals at 2 weeks, persisting in group 4 animals at 6 weeks, but having lysed in controls at 6 weeks. No adhesions were present in group 3 specimens at any interval. Grade I adhesions were present in group 2 at 2 weeks at exposed mesh areas, and declined in frequency at 6 weeks. Evidence of reabsorption of the polyglactin mesh-solder composite was apparent in the group 3 specimens at 4 weeks, and complete resorption had occurred by 6 weeks postoperatively. DISCUSSION: Laser assisted solder fixation caused minimal adhesion formation when mesh was covered by solder. Adhesions were observed if Mersilene mesh material was exposed to the abdominal contents. Vicryl mesh-solder composites reabsorbed without inflammation, scarring, or adhesions at the sites of mesh fixation. CONCLUSION: Further development of this technology is warranted. PMID- 15898899 TI - Is pneumoperitoneum harmful during intra-abdominal hemorrhage in rats? AB - BACKGROUND: Laparoscopic surgical interventions are being used in trauma patients for diagnostic and therapeutic purposes, but there are limited studies on this subject. The effect of pneumoperitoneum during intra-abdominal hemorrhage has not been elucidated. The aim of this study was to investigate the hemodynamic, respiratory, and renal effects of pneumoperitoneum in the splenic injury/ hemorrhagic shock model in rats. MATERIAL AND METHODS: In this study, 80 anesthetized Wistar male rats (294.5 +/- 31.2 g) were randomized into 2 main groups: nontraumatized (group A) and traumatized (group B). After initial preparation and monitoring, each group was divided according to the degree of pneumoperitoneum. The nontraumatized subgroups were A1, sham-operated; A2, 4-8 mm Hg; A3, 9-13 mm Hg; and A4, 14-18 mm Hg. The traumatized subgroups were B1, splenic injury without pneumoperitoneum; B2, B3, and B4, splenic injury with pneumoperitoneum at 4-8 mm Hg, 9-13 mm Hg, and 14-18 mm Hg, respectively. Mean arterial pressure, heart rate, and respiratory rate were monitored continuously. Blood samples were obtained for hemoglobin, hematocrit, arterial blood gases, and biochemical analyses. Twenty-four hour urine output was collected. RESULTS: In group B4, pH, pCO2, and HCO3 levels were lower than in all other groups, while pCO2 and base deficit levels were significantly higher (P < 0.05). Both blood and urine analysis results showed that 24-hour urine output and the glomerular filtration rate of groups A4 and B4 were significantly lower (P < 0.05), while urinary osmolarity and fractional sodium excretion levels were significantly higher (P < 0.05). CONCLUSION: High-pressure pneumoperitoneum in splenically traumatized rats amplifies acidosis, decreases urine output, decreases glomerular filtration rate, and increases urinary osmolarity and fractional sodium excretion significantly. PMID- 15898900 TI - Endoscopic totally extraperitoneal inguinal hernioplasty under spinal anesthesia. AB - OBJECTIVES: Although endoscopic totally extraperitoneal inguinal hernioplasty (TEP) confers superior early outcomes compared to those of open repair, the requirement of general anesthesia has been held as an argument against the application of TEP by opponents of laparoscopic surgery. To date, the literature on TEP performed under spinal anesthesia remains scarce. The present study reports our early experience performing TEP under spinal anesthesia in selected patients who were medically unfit for general anesthesia. METHODS: Between March 2003 and March 2004, 6 male patients underwent attempted TEP under spinal anesthesia. Selection criteria for the procedure included reducibility of the inguinal hernia and concomitant medical conditions precluding general aesthesia, such as impaired lung function. Informed consent was obtained in all patients. RESULTS: All patients were conscious and able to communicate verbally during the operation. TEP was successfully completed in 4 patients, with a mean operative time of 33 minutes. All 4 patients were asymptomatic and experienced no pain throughout the procedure. Conversion to open repair was required in 2 patients because of uncooperative movement in one, and inadequate neural blockade by spinal anesthesia in the other. Intraoperative cardiorespiratory parameters were stable in all patients. Postoperative urinary retention occurred in 1 patient. The mean length of follow-up exceeded 3 months, and no seroma or recurrence was detected clinically. CONCLUSION: Successful performance of TEP under spinal anesthesia requires the combined efforts of an experienced anesthesiologist, a skilled surgeon, and a cooperative patient. Our initial experience of TEP under spinal anesthesia appeared promising. TEP under spinal anesthesia may have a role in selected patients who are medically unfit for general anesthesia but are otherwise suitable for TEP. PMID- 15898901 TI - The effect of low-flow sevoflurane and desflurane on pulmonary mechanics during laparoscopic surgery. AB - BACKGROUND: This study was designed to investigate the effects of inhaled anesthetic agents on respiratory mechanics with low flow anesthesia in laparoscopic abdominal surgery. Two inhaled anesthetics, desflurane and sevoflurane, have a lower solubility in blood and tissues than all previous volatile anesthetics, and have become the preferred volatile anesthetics for routine surgical procedures. METHODS: Twenty-six patients were examined. Patients were randomly assigned to two groups, to receive sevoflurane (n = 13) or desflurane (n = 13). Tidal volume and ventilation rate were kept unchanged throughout the operation. Intra-abdominal pressure was kept constant at the level of 12 mm Hg. Respiratory mechanics such as peak inspiratory pressure (PIP), respiratory resistance (Rr), and dynamic compliance (Cdyn) measurements were recorded by a Datex-Ohmeda respiratory device (Datex-Ohmeda, Finland) at four timepoints: 5 minutes after mechanical ventilation started (T1), after insufflation of the peritoneum (T2), in the 30-degree Trendelenburg position (T3), and after desufflation of the peritoneum (T4). RESULTS: In our study, desflurane caused a statistically significant increase in PIP and Rr and decrease in C(dyn). When the two groups were compared, Rr values in the deslurane group showed significant increases at T2, T3, and T4 compared to the sevoflurane group (P < 0.05). These values did not change in the sevoflurane group, while PIP significantly increased at T2, T3, and T4 after desufflation in the desflurane group (P < 0.05). Cdyn values decreased significantly at all 4 timepoints in the desflurane group compared to the sevoflurane group (P < 0.05). CONCLUSION: We concluded that respiratory mechanics were affected by desflurane with low flow anesthesia in patients undergoing laparoscopic abdominal surgery. No significant influence on respiratory mechanics was seen with sevoflurane anesthesia. PMID- 15898902 TI - The effect of the degree of histologic inflammation on gallbladder perforation during laparoscopic cholecystectomy. AB - PURPOSE: Laparoscopic cholecystectomy (LC) is the gold standard operation for the treatment of symptomatic gallstones. Intraoperative gallbladder perforation is a common complication encountered during the surgery. The purpose of this study was to identify the effects of inflammation on gallbladder perforation during LC. METHODS: Between July 1997 and March 2003, 509 patients underwent LC for symptomatic gallstone disease at the Department of Surgery at Vakif Gureba Training Hospital. Data were collected retrospectively. Patients with and without gallbladder perforation were compared in terms of gender, age, anatomic difficulty, experience of the surgeon, omental and other organ adhesions to the gallbladder, and the findings of inflammation on the resected gallbladder. RESULTS: Intraoperative gallbladder perforation occurred in 85 patients (16.6%). Although no differences were found between the perforated and nonperforated groups regarding age, gender, and chronic inflammation on the resected gallbladder, there were significant differences in terms of acute inflammation, anatomical difficulty, experience of the surgeon, and omental and organ adhesions. CONCLUSION: Acute inflammation, degree of anatomic difficulty, the experience of the surgeon, and omental or other organ adhesions were associated with gallbladder perforation during LC. PMID- 15898903 TI - Laparoscopic versus open splenectomy in the management of benign and malign hematologic diseases: a ten-year single-center experience. AB - OBJECTIVE: Laparoscopic splenectomy (LS) has gained wide acceptance as a safe, effective alternative to open splenectomy (OS) in the treatment of hematologic disorders. The aim of this study was to compare two cohorts of patients with similar characteristics, who underwent LS and OS in a single university teaching center. METHODS: Records were reviewed from 30 patients who underwent LS for a hematologic disease and compared with a control group of 38 patients undergoing OS for hematologic disease. Demographics and outcomes were recorded. RESULTS: There were no significant differences between the two groups with respect to accessory spleens, blood loss, or complication rates (P > 0.05). The operation time in the LS group was significantly longer than in the OS group (P < 0.01) and the length of hospital stay in the LS group was significantly shorter than in the OS group (P < 0.01). CONCLUSION: Laparoscopic splenectomy is likely becoming the gold standard in the surgical treatment of hematologic diseases. PMID- 15898904 TI - The lithotomy versus the supine position for laparoscopic advanced surgeries: a historical review. AB - Laparoscopic advanced surgery has been taught in many institutions in the United States. Initially, proctoring for the laparoscopic technique was performed by European surgeons; therefore, the lithotomy position was suggested as the preferred approach. Many American and European surgeons have adopted the supine position. Laparoscopy initially entered the clinical realm in the field of gynecology. Albert Decker, at the Knickerbocker and Gouverneur Hospital in New York, performed culdoscopy as early as 1928. This was done in the "knee-chest" position without the use of pneumoperitoneum. Raoul Palmer, at the Hopital Broca in Paris, popularized "colposcopie," utilizing pneumoperitoneum, with the patient in the lithotomy position. Laparoscopy then advanced in Europe to the general surgery arena. As a result, patient positioning for laparoscopic procedures in Europe was performed in what is now referred to as the French position (i.e., lithotomy). Many of these procedures are modified to a side approach, or American position, when performed in the United States. There is a clear association between the dorsal lithotomy position and the development of postoperative compartment syndrome. Compartment syndrome occurs when elevated pressure in an osteofascial compartment compromises local perfusion, and often results in neurovascular damage and permanent disability. Many centers have adopted the lithotomy position for their laparoscopic advanced procedures. At our institution, however, we prefer all procedures be performed in the American position (patient supine and the surgeon at the side of the patient), since this resembles the position used for other, open surgeries. The advantage of this approach is that it eliminates the risks associated with placement of the patient in the lithotomy position. PMID- 15898905 TI - Simulated hand-assisted laparoscopic surgery (HALS) in microgravity. AB - INTRODUCTION: Previous simulation and porcine experiments aboard the reduced gravity program KC-135 turbojet have demonstrated that microgravity surgery is feasible. Ideally, surgical care in spaceflight will incorporate recent advances in care while remaining easy enough for a crew medical officer (CMO) lacking surgical proficiency or extensive surgical experience to perform. As a minimally invasive surgical technique, hand-assisted laparoscopic surgery (HALS) benefits the patient via smaller incisions, less pain, and faster recovery than traditional open surgery. HALS also helps less experienced laparoscopic surgeons perform laparoscopic surgery. METHODS: An inexpensive inanimate surgical simulator was constructed to evaluate the usefulness of HALS in microgravity. This simulator was utilized during brief periods of microgravity provided by parabolic flight on the KC-135. The simulator was successfully used by both a physician-astronaut and an experienced laparoscopic surgeon. Task completion included simulated surgery with exploration of the intestines and ligation of the appendix. RESULTS: Simulated HALS was successfully performed in microgravity. HALS effectively contained operative equipment and small amounts of introduced fluids within the simulated abdominal cavity. Astronaut and surgeon experience suggest that HALS could facilitate minimally invasive surgery (MIS) in microgravity. DISCUSSION: HALS holds promise as a surgical approach in microgravity, particularly as space travel extends beyond low earth orbit. HALS provides the benefits of MIS, facilitates MIS surgery by less surgically proficient or experienced CMOs, and contains equipment and fluid within the operative field. Simulation provides an easy, cost-effective platform to evaluate medical technology for space flight as well as a method to train CMOs on-orbit. PMID- 15898906 TI - Laparoscopic ureterolithotomy: the method of choice in selected cases. AB - OBJECTIVE: To evaluate retrospectively the results and complication, efficacy, and safety rates of laparoscopic ureterolithotomy in the treatment of large, impacted ureteric stones. MATERIALS AND METHODS: Between September 1994 and December 2003, 73 patients underwent 75 laparoscopic ureterolithotomies (LUL). Mid- and upper-ureteric stones were removed retroperitoneally in 69 cases (92%). Lower ureteric stones were removed transperitoneally in 6 cases (8%). There were 2 (3%) bilateral LULs in one session, 7 (10%) ipsilateral percutaneous nephrolithotomy, and 6 (8%) ipsilateral ureteroscopy in the same session. The mean age of the patients was 39 years (range, 15-70 years). Mean stone size was 25 mm (range, 12-55 mm). RESULTS: The success rate was 98.7%, with 1 patient (1.3%) requiring conversion to an open procedure. There were no major complications. Mean operative time was 45 minutes (range, 15-100 minutes). Postoperative analgesic medication was required for 1 day in 39 patients (52%). The postoperative hospital stay ranged from 2 to 5 days (mean, 3 days). CONCLUSION: On the basis of our experience, laparoscopic ureterolithotomy is a safe and effective method for the treatment of large, dense, and impacted ureteric stones. In selected cases it can be the first choice of treatment. PMID- 15898907 TI - Laparoscopic surgery for the management of obstruction of the gastric outlet and small bowel following previous laparotomy for major upper gastrointestinal resection or cancer palliation: a new concept. AB - BACKGROUND: Surgical relief of gastric outlet obstruction (GOO) or small bowel obstruction in patients who had undergone major resection or palliative bypass surgery for malignancy is conventionally achieved at a laparotomy. The potential role of minimally invasive surgery in the management of these complications has not been previously explored. METHODS: Between 2003 and 2004, 4 consecutive patients, age range 37 to 72 years, where admitted with gastric outlet or proximal small bowel obstruction following previous open surgery for suspected intra-abdominal malignancy, under the care of one surgeon. The respective past histories of these patients were recurrent GOO and concomitant distal biliary obstruction following a previous open gastric bypass elsewhere for metastatic pancreatic head cancer; persistent adhesive small bowel obstruction following radical gastrectomy for gastric cancer; GOO secondary to intra-abdominal recurrence 6 months after hepatobiliary resection for hilar cholangiocarcinoma; and GOO following previous pancreatico-duodenectomy for suspected pancreatic head cancer. Their respective surgical management consisted of a laparoscopic re-do gastric bypass and concomitant cholecystojejunostomy; adhesiolysis and revision of the Roux-en-Y enteric anastomosis; a Devine exclusion gastroenterostomy; and resection and refashioning of the gastroenterostomy. RESULTS: There were no conversions to open surgery and no postoperative complications. The median operating time was 240 minutes (range, 145 to 300 minutes). Oral free fluid intake was resumed on postoperative day (POD) 1, while diet was resumed between POD 2 and 4. The median postoperative hospital stay was 15.5 days (range, 14 to 25 days). CONCLUSION: Previous laparotomy and major resection or palliation of malignancy do not preclude the application of the laparoscopic approach for the management of upper gastrointestinal obstruction. Laparoscopic adhesiolysis and revision of enteroenteric and gastroenteric anastomoses are feasible management options in the hands of those experienced with complex laparoscopic surgery. PMID- 15898908 TI - Laparoscopic splenectomy for isolated sarcoidosis of the spleen. AB - Sarcoidosis is a granulomatous disease of unknown etiology. Over 90% of patients with sarcoidosis present with pulmonary findings at the time of diagnosis. Extrapulmonary involvement is common, including the liver, eyes, central nervous system, lymph nodes, and joints. However, isolated granulomatous disease confined to the spleen is rare. This report documents a rare case of isolated granulomatous disease of the spleen diagnosed and treated laparoscopically. A 47 year-old female presented to her internist with nausea and mild epigastric abdominal pain. Laboratory evaluation revealed slightly elevated bilirubin and liver function tests as well as neutropenia. An abdominal ultrasound revealed normal biliary and pancreatic anatomy and multiple splenic lesions. Computed tomography of the abdomen confirmed the multiple hypodense lesions within the spleen. The differential diagnosis at this time included neoplasm, infection, and autoimmune etiologies. Subsequently, the patient underwent a diagnostic laparoscopy with splenectomy. No other intra-abdominal pathology was found. Pathology revealed multiple noncaseating, splenic granulomas. Isolated extrapulmonary manifestations of sarcoidosis occur in only 10% of these patients. Moreover, isolated splenic sarcoidosis is rare. Currently, the literature documents only two prior cases of sarcoidosis presenting with isolated splenic lesions. Primary management consists of medical therapy with prednisone, methotrexate, and/or antimalarial drugs. Indications for surgery include symptomatic splenomegaly, severe hypersplenism, prophylaxis for splenic rupture, and neoplastic exclusion. Our patient required a laparoscopic splenectomy for diagnostic purposes and neoplastic exclusion. Once diagnosed, patients require continual follow-up for systemic manifestations and associated complications of sarcoidosis. PMID- 15898909 TI - Bronchogenic cyst of the gastric fundus presenting as a gastrointestinal stromal tumor. AB - Gastrointestinal stromal tumors (GIST) of the stomach are being recognized with increasing frequency. The diagnosis is usually made on the basis of appearance on computed tomography (CT) and excision is recommended for GIST larger than 5 cm. We report a 39-year-old woman referred for resection of a presumed GIST of the gastric fundus diagnosed by CT scan and upper gastrointestinal (UGI) series. A laparoscopic resection was performed, but upon pathologic examination the mass proved to be a bronchogenic duplication cyst of the stomach. Bronchogenic and esophageal duplication cysts usually arise in the chest or mediastinum. On rare occasions bronchogenic cysts may lose their connection to the tracheobronchial tree and migrate to a subcutaneous position in the neck or descend into the retroperitoneum. The importance of this case is that it demonstrates a rare yet essential component to the differential diagnosis of lesions of the stomach. PMID- 15898910 TI - Agenesis of the right liver: a difficult laparoscopic cholecystectomy. AB - Congenital agenesis of the right liver (ARL) is a rare condition that is generally asymptomatic. Congenital anomalies of the liver are generally diagnosed with current cross-sectional imaging modalities such as ultrasound, computed tomography, and magnetic resonance imaging (MRI) before surgery. However, discovery of a congenital anomaly of the liver only at time of surgery remains a possibility. Herein reported is the case of a 68-year-old woman undergoing laparoscopic cholecystectomy for symptomatic gallbladder stones as diagnosed on preoperative ultrasounds. Upon laparoscopic exploration of the upper abdomen, the right liver was not found; the gallbladder was located in the right subdiaphragmatic region posterior to the medial segment of the liver. A posterolateral interposition of the hepatic flexure of the colon was also found. Cholecystectomy was completed under laparoscopy. A postoperative MRI confirmed right liver agenesis. We discuss the technical difficulties of performing a laparoscopic cholecystectomy in the case of ARL and the advantages of a laparoscopic approach. PMID- 15898911 TI - Transanal endoscopic repair of rectal perforation with hemoclips. AB - A 74-year-old male patient had a full-thickness laceration of the mid-rectum during transurethral urologic surgery. The perforation site was 8-cm above the dentate line and 15 x 6 mm in size. Transanal direct access to the injury site was difficult and endoscopic repair with hemoclips was performed without any further intervention. He was discharged from the hospital uneventfully and complete healing was observed 6 weeks after the procedure. We concluded that endoscopic repair with hemoclips may offer a successful and simple closure method for extraperitoneal high lying rectal lacerations. PMID- 15898912 TI - Laparoscopically assisted resection of an ascending colon lipoma causing intermittent intussusception. AB - Colonic lipomas are relatively uncommon clinical entity, although they are among the most common nonepithelial benign tumors of the colon. They may produce abdominal pain, diarrhea, constipation, hemorrhage, and intussusception. We report a case of a 56-year-old man who was suffering from abdominal pain and 4 weeks of intermittent diarrhea. Ultrasound examination of the abdomen revealed an abdominal mass in the right upper quadrant. Computed tomography showed a well circumscribed intraluminal mass in the ascending colon, measuring 5 cm in diameter, with density value equal to fat. During laparoscopic surgery we found a colocolic intussusception, which we reduced laparoscopically, and performed a laparoscopically assisted right colectomy. The resected colon revealed a 6 cm wide lipoma within the ascending colon. Histology confirmed the diagnosis of benign lipoma. Aspects of preoperative diagnosis and treatment are discussed, and we review the literature. PMID- 15898913 TI - Disposable laparoscopic surgical instruments and the economic effects of repackaging. AB - The purpose of this study was to evaluate the economic effects of repackaging disposable laparoscopic surgical instruments. We repacked a total of 28 disposable instruments for gynecologic laparoscopic surgery into one package. The time of preparation of surgical instruments, the amount of medical waste, and other economic effects were compared before and after the repackaging of disposable instruments. The time required for the preparation of surgical instruments was shortened by 22 minutes per operation, and medical waste decreased by 400 g, by repackaging the instruments, resulting in an estimated direct economic saving of $10,000 per year. Indirect economic effects might include savings due to a reduced inventory of surgical instruments and simplified cost-accounting for each operation. It is likely that preparation for surgery can be significantly improved and the cost reduced by repackaging the laparoscopic surgical instruments. PMID- 15898914 TI - Computer-assisted, robot-enhanced open microsurgery in an animal model. AB - BACKGROUND: Computer-assisted, robot-enhanced surgery improves laparoscopic and thoracoscopic surgery through tremor filtration, motion scaling, articulation, and improved ergonomics. Surgeons perform many open cases under magnification that magnifies the tremor present in all surgeons' hands, so the tremor filtration and motion scaling of robotic surgery may improve microsurgery. Our goal was to compare microvascular anastomoses performed with a robot-enhanced technique with a standard technique. METHODS: We performed end-to-end anastomoses in 1-mm rat femoral arteries with interrupted 10-0 suture. We compared the anastomotic time, patency, and leak rates between traditional microsurgery techniques (by hand) and a robot-enhanced technique using the Zeus robotic surgery system (Computer Motion, Goleta, California). The surgeon used an operative microscope for visualization in both techniques. RESULTS: We performed 30 anastomoses by hand and 31 with Zeus. We observed a remarkable degree of tremor filtration in the robot-enhanced cases. Anastomotic times for both techniques demonstrated a learning curve. Anastomoses done by hand (mean time, 17.2 minutes) were significantly faster than those done with Zeus (mean time, 27.6 minutes) (P = 0.0006). All anastomoses from both groups were patent, and none leaked after 3 minutes. CONCLUSION: The Zeus system is effective at performing complex, open, microsurgery tasks in vivo. There was no measurable benefit from the remarkable tremor filtration and motion scaling offered by robot enhanced surgery. PMID- 15898915 TI - Elective laparoscopic appendicectomy for chronic right iliac fossa pain in children. AB - AIM: To determine whether elective laparoscopic appendicectomy is justified for chronic right iliac fossa (RIF) pain of undetermined origin. METHODS: A retrospective audit of all laparoscopic appendicectomies between January 1997 and August 2003 was performed. The expanded medical audit system (EMAS) and a Microsoft Access database of operative records were used to identify patients with chronic RIF pain subjected to elective appendicectomy. Case notes were retrieved and analysed for patient profile, duration of symptoms including clinic visits and admissions, operative findings, histological analysis, and postoperative performance. A correlation between histological findings and final outcome was investigated. RESULTS: Ninety-eight patients underwent laparoscopic appendicectomy during the period of the study. A total of 11 cases with chronic RIF pain were identified. Nine were female and 2 male. Age ranged from 9 to 14 years with a mean of 11.9 years. The number of clinic visits and admissions for chronic RIF pain ranged from 2 to 8, with a mean of 4. Duration of symptoms ranged from 1 to 36 months, with a mean of 12.1 months. Detailed history, clinical examination, and serological and radiological investigations failed to reveal the cause of the pain in all cases. Patients were followed up in postoperative clinics for between 1 and 72 months, with a mean of 16.1 months. Histology of resected appendices showed acute inflammation (3 cases), fecoliths (2 cases), lymphoid hyperplasia (LH) (1 case), LH and a foreign body reaction (1 case), LH and mucosal hyperplasia (1 case), and Enterobius vermicularis parasites in 1 case. The appendix was normal in 2 cases. Eight patients had complete resolution of RIP pain. Seven of these had pathology within the appendix and 1 was histologically normal. Two patients with resolved RIF pain, but with pain elsewhere, had lymphoid hyperplasia noted within the appendix. One patient with persistent pain 6 years postoperatively had a normal appendix. CONCLUSION: This study demonstrates that a significant number of patients with chronic RIF pain have pathology within the appendix. The majority of these cases will benefit from elective appendicectomy. It is critical however that all other possible causes of pain in the RIF are excluded. Laparoscopy is an integral part of the diagnosis and management of this particularly difficult group of patients. PMID- 15898916 TI - Outcome and cost analysis of laparoscopic or open surgery versus conservative management for multicystic dysplastic kidney. AB - PURPOSE: To determine whether laparoscopic nephrectomy (LN), open nephrectomy (ON), or observation (OB) is most efficient for managing multicystic dysplastic kidney (MCDK). METHODS: We performed a retrospective review of the management of our 12-year clinical experience of 32 MCDK patients to analyze the estimated total cost (ETC) of various treatment options and compare them with respect to survival, development of hypertension, and postoperative cosmetic appearance, to determine the most efficient treatment option. RESULTS: There were 12 cases in the LN group, 6 in the ON group, and 14 in the OB group. ETC was lower for ON than for LN. But the length of preoperative observation and length of follow-up after spontaneous regression greatly affected ETC. If MCDK cases were observed > 5 years with standard investigations performed regularly, ETC was higher for the OB group than for the LN group. In 6/14 (42.9%) of OB cases, MCDK resolved within 5 years, and this subgroup had the lowest ETC up to the time of this review. Six cases initially in the OB group were managed surgically (4 by LN and 2 by ON) in accordance with parental requests, and ETC in these cases was highest. All cases are well after a mean follow-up of 5.7 years, without any impairment of renal function, or hypertension. CONCLUSION: The decision to operate after lengthy observation increases costs. Overall, the most efficient surgical technique for managing MCDK was LN because of greater patient satisfaction with postoperative cosmesis. We recommend MCDK be treated by LN after a short period of observation. PMID- 15898917 TI - Endoscopic resection of a juvenile angiofibroma: the role of the XPS microdebrider. AB - Juvenile angiofibromas are vascular tumors found almost exclusively in the adolescent male. Although benign, their clinical course can be aggressive and can result in major morbidity and mortality. Endoscopic transnasal resection of juvenile angiofibromas, confined to the nose, paranasal sinuses, pterygopalatine fossa, and medial infratemporal fossa, has been a significant advance in their management, eliminating or reducing the need for extensive soft tissue and bony dissection of traditional surgical approaches. We discuss a case of a juvenile angiofibroma resected transnasally using the XPS microdebrider (Medtonic Xomed, Jacksonville, Florida) and outline the role of this instrument in this surgery. PMID- 15898918 TI - Laparoscopic femoral hernia repair using umbilical ligament as plug. AB - A novel technique for the laparoscopic repair of femoral hernia is described. An 11-year-old boy who had undergone previous open inguinal herniorrhaphy presented with a persistent bulge in the ipsilateral groin. At laparoscopic exploration, a small femoral hernia was discovered. It was reduced with extrinsic compression and the defect closed laparoscopically using a permanent purse-string suture. The ipsilateral umbilical ligament was dissected off the abdominal wall and secured (plugged) over the internal opening, providing a tension-free repair. The immediate postoperative result was satisfactory and there was minimal postoperative pain. Two-year follow-up confirmed adequate repair, without recurrence. This laparoscopic technique enabled diagnostic precision, contralateral evaluation, and a novel means of patching and buttressing the peritoneal closure with umbilical ligament. The procedure is safe, simple, and provides excellent functional and cosmetic results. PMID- 15898924 TI - Digital volume tomography in the diagnosis of periodontal defects: an in vitro study on native pig and human mandibles. AB - BACKGROUND: The aim of this study of native pig and human mandibles was to investigate the accuracy and quality of the representation of periodontal defects by intraoral radiography (IR), panoramic radiography (PR), computed tomography (CT), and digital volume tomography (DVT) in comparison with histologic specimens. METHODS: Following the standardized preparation of periodontal defects (14 dehiscences, fenestrations, 2- to 3-walled intrabony defects, respectively; Class I, II, and III furcation involvement) in six pig and seven human mandibles, IR, PR, CT, and DVT were performed. The histologic specimens were produced by cutting blocks with the individual defects out of the mandibles, embedding them in acrylic, and producing sagittal and axial microsections. The intrabony defects were measured using appropriate software on the digitized IR and PR images programs. The histologic sections were measured by reflecting stereomicroscopy. The statistical comparison between the measurements of the radiographic images and those of the histologic specimens was performed with Pearson's correlation coefficient. The quality of the radiographic images was determined through the subjective perception and detectability of the intrabony defects by five independent observers. RESULTS: All intrabony defects could be measured in three planes in the CT and DVT scans. Comparison with the histologic specimens yielded a mean deviation of 0.16 +/- 0.10 mm for the CT scans and 0.19 +/- 0.11 mm for the DVT scans. On the IR and PR images, the defects could be detected only in the mesio-distal and craniocaudal planes. In comparison with the histologic specimens, the IR images revealed a mean deviation of 0.33 +/- 0.18 mm and the PR images a mean deviation of 1.07 +/- 0.62 mm. The quality rating of the radiographic images was highest for the DVT scans. CONCLUSIONS: Overall, the CT and DVT scans displayed only a slight deviation in the extent of the periodontal defects in comparison with the histologic specimens. Both radiographic imaging techniques permitted imaging of anatomic osseous structures in three planes, true to scale, and without overlay or distortion. The DVT scans showed the best imaging quality. PMID- 15898925 TI - Distribution of Porphyromonas gingivalis fimA genotypes in Japanese children and adolescents. AB - BACKGROUND: Porphyromonas gingivalis is considered to be one of the most important pathogens in periodontal disease and its fimA genes have been classified into six variants (types I through V and Ib). A majority of the P. gingivalis organisms isolated from periodontitis patients are type II, followed by type IV, while type I is prevalent in healthy adults. METHODS: A total of 650 saliva samples were taken from 464 children (3 to 18 years of age), who visited Osaka University Dental Hospital. Genomic DNA was extracted from each sample and analyzed using a polymerase chain reaction (PCR) method with P. gingivalis specific primers, followed by an additional PCR assay to determine the fimA genotypes for P. gingivalis- positive subjects. RESULTS: Fifteen (3.23%) of the subjects were P. gingivalis-positive and none of those samples showed a positive reaction to the type II fimA-specific primers, while four, one, and two subjects were shown to be positive for the type I, Ib, and III genotypes, respectively. In addition, the type IV genotype was detected in three subjects in the older age group. CONCLUSIONS: Our findings suggest that a limited number of children harbor P. gingivalis, and that the distribution of type II and IV fimA genotypes is extremely low. Further, some adolescents were found to possess the type IV fimA genotype which has been shown to be possibly related to adult periodontitis, in contrast to types I, III, and V. PMID- 15898926 TI - The trans-chromosomic mouse-derived human monoclonal antibody promotes phagocytosis of Porphyromonas gingivalis by neutrophils. AB - BACKGROUND: As a safe immunotherapeutic approach, human monoclonal antibody (hMAb) may be effective in clearing periodontopathic bacteria. The trans chromosomic (TC) technology has recently been applied to construction of the TC mouse, which enables us to incorporate entire human chromosome fragments containing immunoglobulin (Ig) gene cluster. The aim of this study is to establish TC mouse-derived hMAb, and to test the in vitro opsonophagocytic activity. METHODS: Human Ig-producing TC mouse was immunized by recombinant 40 kDa outer membrane protein (r40-kDa OMP) of Porphyromonas gingivalis 381, and the spleen cells were fused with the mouse myeloma cell line. The specificity of antir40- kDa OMP hMAb was evaluated with the enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance assays. Flow cytometric analyses were performed to assess the opsonophagocytic activity. RESULTS: We successfully constructed 99 IgG isotype clones (IgG1: 84; IgG2: 11; IgG4: four clones), which were specifically reactive with r40-kDa OMP. The anti-r40-kDa OMP IgG1 hMAbs promoted phagocytosis of P. gingivalis by neutrophils. Futhermore, an increased opsonophagocytic activitity of anti-r40-kDa OMP IgG1 hMAbs was observed not only in P. gingivalis 381, but also in the W50, W83, and Su63 strains. CONCLUSION: Our results document the TC mouse-derived hMAb to promote neutrophil phagocytosis of P. gingivalis, suggesting an immunotherapeutic option for clearance of P. gingivalis. PMID- 15898927 TI - Er:YAG laser in defocused mode for scaling of periodontally involved root surfaces: an in vitro pilot study. AB - BACKGROUND: The Er:YAG laser may be used on periodontally involved teeth in combination with conventional periodontal therapy in order to improve the efficacy of root instrumentation. The aim of this study was to compare the effect of hand instrumentation on root surfaces of periodontally involved teeth with Er:YAG laser application. METHODS: Thirty freshly extracted, non-carious, single rooted, periodontally diseased human teeth from adult humans with advanced periodontal disease were used in this study. The teeth were divided into three groups of 10 specimens each. Group A was treated with scaling and root planing (SRP) with curets only (control). In group B, the root surfaces were scaled with curets and then lased with an Er:YAG laser (wavelength 2.94 microm). A handpiece with a water spray was used in non-contact mode (defocused) at a distance of 1 cm from root surface. Laser parameters were set at energy of 100 to 200 mJ/pulse, with 10 Hz of frequency. In group C, the root surfaces were lased only with power settings 250 to 300 mJ/pulse and 10 Hz frequency. An epon-araldite plastic embedding technique was used for light microscopic investigation. RESULTS: Histologic findings showed significant differences between the test and control sites. In control sites, after hand instrumentation, the surface was smooth, without a cementum layer, and the dentin layer presented opened tubules. Defects on the dentin layer were also present along root surfaces. In the test sites (B, C) root surfaces revealed no thermal damage; no cracking or tissue carbonization were observed. The superficial layers of lased surfaces appeared smooth and melted without alterations. CONCLUSION: Based on these findings, it appears that it may be feasible to use the Er:YAG laser for root instrumentation without prior root planing if the proper parameters are followed. PMID- 15898928 TI - Effect of cyclosporin A on apoptosis and expression of p53 and bcl-2 proteins in the gingiva of renal transplant patients. AB - BACKGROUND: Gingival overgrowth (GO) is a common side effect of cyclosporin A (CsA) therapy, but the exact mechanism for this is unknown. Apoptosis plays an important role in the maintenance of tissue homeostasis and mediators of this process may be involved in the pathogenesis of drug-induced GO. This study compared p53 expression, bcl-2 expression, and apoptosis in gingival samples from CsA-treated renal transplant recipients to findings in controls with gingivitis. METHODS: Twenty-two kidney recipients with CsA-induced GO and 15 systemically healthy subjects with gingivitis were included in the study. The 15 systemically and periodontally healthy volunteer control group were immunohistochemically analyzed for grades of p53 and bcl-2 expression, and were processed using terminal TdT-mediated dUTP-biotin nick-end labeling (TUNEL) technique to identify and grade levels of apoptosis. RESULTS: There were no differences between the CsA group and the control group with respect to grades of p53 and bcl-2 expression (P >0.05 for both). However, the CsA group showed a lower apoptosis grade than the control group (P <0.05). None of the clinical parameters was significantly correlated with any of the immunohistochemical findings for p53 or bcl-2 (P >0.05 for all). Similarly, grade of apoptosis was not correlated with any of the clinical parameters (P >0.05). There was a significant positive correlation between serum CsA level and level of bcl-2 expression, but serum CsA was not significantly correlated with level of apoptosis or level of p53 expression. CONCLUSION: The results indicate that the pathogenesis of CsA-induced GO might involve inhibition of apoptosis, and overexpression of bcl-2 in the setting of high serum CsA. PMID- 15898929 TI - Syndecan-1 expression during the formation of junctional epithelium. AB - BACKGROUND: Syndecans are cell surface heparan sulfate proteoglycans (PG) which can bind to and modulate the action of growth factors and extracellular matrix components (ECM). Syndecan- 1 has been shown to play important roles during early tooth development and wound healing and repair. Among diverse cells and tissues that comprise the periodontium, the junctional epithelium (JE) constitutes a region of significant anatomic and clinical importance, but the nature of inductive signals and molecules involved in its formation is still unclear. Therefore, this work examines if syndecan-1 is associated with formation of JE, and the distribution of other syndecan family members in the epithelium. METHODS: In situ hybridization and immunohistochemical techniques were performed using oral tissues from 4-day-old to 10-week-old mice to investigate the expression of syndecan- 1, -2, -3 and -4 mRNAs and their corresponding proteins. RESULTS: Based on in situ hybridization experiments, all syndecan mRNAs were detected in sulcular epithelium (SE), gingival epithelium (GE), and JE with varying intensity and distribution. Syndecan-1 immunostaining was localized on the cell surface while that of syndecan-2 did not show clear membrane localization. Our experiments in the developing tooth demonstrated that syndecan-1 protein followed characteristic patterns of expression during JE formation and that immunoreactivity for syndecan-1 protein decreased with age when JE cells underwent terminal differentiation. CONCLUSION: Results of syndecan-1 mRNA and protein expression patterns suggested that this proteoglycan might be an important molecule during the formation of JE. PMID- 15898930 TI - The impact of periodontal therapy and the adjunctive effect of antiseptics on breath odor-related outcome variables: a double-blind randomized study. AB - BACKGROUND: Bad breath is often caused by periodontitis and/or tongue coating. This study followed the impact of initial periodontal therapy on several halitosis-related outcome variables over a 6-month period. Organoleptic ratings are often uncomfortable for the patient and have several disadvantages. They are, for instance, influenced by external parameters (e.g., food intake and cosmetics) and need to be calibrated among researchers worldwide. A second aim was to evaluate the reliability of saliva incubation as an in vitro indirect test for breath recording. METHODS: In this double-blind, randomized, medium-term, parallel study 45 moderate periodontitis patients without obvious tongue coating were enrolled. Besides a one-stage, full-mouth disinfection and oral hygiene improvement (including daily tongue scraping), patients were instructed to rinse daily for 6 months with one of the following products (randomly allocated): chlorhexidine (CHX) 0.2% + alcohol, CHX 0.05% + cetyl pyridinium chloride (CPC) 0.05% without alcohol (a new formulation), or a placebo solution. At baseline and 3 and 6 months, a series of parameters were recorded including: concentration of volatile sulfide compounds (VSC), tongue coating, and an estimation of the microbial load (at anterior and posterior parts of the tongue, saliva, dental plaque). The intraoral VSC ratings were compared to in vitro VSC recordings and organoleptic evaluations of the headspace air from 1 and 2 hours incubated saliva (0.5 ml, 37 degrees C, anaerobic chamber). RESULTS: Even though the initial VSC values were not high (+/-90 ppb with only 18 patients revealing more than 100 ppb), significant (P <0.05) reductions could be achieved in the CHX and CHX + CPC group, and to a lower extent in the placebo group (P = 0.10). Tongue scraping resulted in a significant reduction (P < or =0.05) of the tongue coating up to month 6 in the placebo and CHX + CPC group, but not in the CHX group (confusion due to staining). The CHX and CHX + CPC group showed, in comparison to baseline, significant (P <0.001) reductions in the number of anaerobic species in the supragingival plaque, in the saliva, and on the anterior part of the tongue. For the posterior part of the tongue the microbial changes remained < or =0.3 log values (P >0.05). For the placebo group, the microbial changes never reached a level of significance (< or =0.3 log values). A strong correlation was found between the intraoral VSC ratings and the 1-hour (r = 0.48, P <0.0001; r = 0.54, P = 0.0003 for baseline data only) and 2-hour (r = 0.43, P <0.0001) VSC production of incubated saliva. The latter also correlated very strongly (r = 0.71) with the number of anaerobic species in the saliva. The VSC values and organoleptic ratings of the incubated saliva also correlated strongly with each other (r = 0.64 for 1-hour and 0.73 for 2-hour incubation). CONCLUSIONS: The results of this study indicate that in patients with moderate periodontitis, initial periodontal therapy including tongue scraping did not have a significant effect on the microbial load of the tongue and had only a weak impact on the VSC level, except when combined with a mouthrinse. Saliva incubation can be used as an indirect way to score breath odor. It offers simplicity, objectivity, and is less invasive. PMID- 15898931 TI - Coronally advanced flap: the post-surgical position of the gingival margin is an important factor for achieving complete root coverage. AB - BACKGROUND: An assessment of the factors affecting the clinical outcome of root coverage procedures may be useful in clinical decision making. The aim of this study is to investigate whether the post-surgical location of gingival margin relative to the cemento-enamel junction (CEJ) can influence the recession reduction (RecRed) and complete root coverage (CRC) following coronally advanced flap procedure (CAF). METHODS: Sixty patients, aged between 22 and 57 years, 15 males and 45 females, each showing maxillary buccal recessions (> or =2 mm) identified as Miller's Class I were consecutively enrolled. All the defects were treated by CAF procedure from 2000 to 2003 by a single operator with more than 20 years of clinical experience. Age, gender, smoking habits, and type of tooth of each patient were recorded. In addition, the following clinical data were measured or computed: recession depth, width of keratinized tissue, probing depth, distance between incisal margin (IM) and CEJ, dental hypersensitivity, clinical attachment level, distance between IM and gingival margin (IMGM), distance between IM and mucogingival junction (IMMG), and the location of gingival margin relative to the CEJ following CAF procedure (GM(1)). A multiple linear regression, and a logistic linear regression analyses were performed. RESULTS: The recession depth at baseline (Rec(T0)) and the location of the gingival margin after suturing (GM(1)) are positively correlated to recession reduction. Complete root coverage appeared to be influenced by GM(1): the more coronal the level of the gingival margin after suturing (GM(1)), the greater the probability of CRC. CONCLUSION: The location of the gingival margin relative to the cemento-enamel junction following CAF procedure seems to affect CRC. PMID- 15898932 TI - Root curvature: differences among dental morphotypes and modifications after mechanical instrumentation. AB - BACKGROUND: This study was designed to describe a method for measuring root curvature of extracted teeth, compare root curvature of four different dental morphotypes before mechanical instrumentation, and measure root curvature after mechanical instrumentation. METHODS: Fifty-five maxillary (14 central incisors, 13 lateral incisors, 14 cuspids, 14 premolars) extracted teeth were analyzed. Measurements were recorded on the horizontal plane at the most apical point of the cemento-enamel junction (CEJ). A 4 mm wide root portion on the buccal aspect was selected and recorded on this plane. A digital scanner traced the circle coinciding with the profile of this root portion and calculated its radius, arc, chord, and arrow (i.e., the perpendicular bisector of the chord). Since root curvature is the inverse of the radius (1/r), the radius was related to root curvature; the arc to the mesio-distal dimension of the root portion; and the reduction of the arrow to root flattening after mechanical instrumentation. The measurements were recorded four times: before treatment (baseline), after polishing, after a first root planing, and after a second root planing. The radii of each dental morphotype (upper central incisors, upper lateral incisors, upper canines, upper premolars) recorded in the first measurement were calculated and used for the statistical analyses: one-way analysis of variance (ANOVA) test and the Tukey multiple comparison method were used to study the curvature. The measurements of the radius, the arc, and the arrow at the four times were used for the statistical analyses: the two-way ANOVA test and the Tukey multiple comparison method were applied in the study of the root modifications after mechanical instrumentation. The accuracy and reliability of the method were also evaluated. RESULTS: The mean radii of the four dental morphotypes were: central incisors: 3.613 +/- 0.258 mm; lateral incisors: 2.558 +/- 0.256 mm; canines: 2.822 +/- 0.238 mm; and premolars: 2.321 +/- 0.179 mm. The statistical analyses revealed differences among central incisors, canines, lateral incisors, and premolars. There was no statistically significant difference between lateral incisors and premolars. Regarding the root modifications after mechanical treatment, the radius did not show statistically significant differences in any of the comparisons. The arc and the arrow did not show significant differences between baseline and polishing, while they did show significant differences after the second root planing. CONCLUSIONS: This study indicates that: 1) the method of measuring is accurate and reliable; 2) there are statistically significant differences among the root curvatures of different dental morphotypes; and 3) polishing did not modify the root. A vigorous root planing did not modify root curvature, but it did reduce the mesio-distal dimension and flatten the root surface slightly. PMID- 15898933 TI - Identification of periodontal pathogens in atherosclerotic vessels. AB - BACKGROUND: Epidemiological studies have shown that periodontitis may be associated with presence of atherosclerosis. DNA from periodontal pathogens has been detected in atherosclerotic lesions, but viable oral bacteria have not yet been isolated from atherosclerotic plaques. The purpose of the present study was to determine if viable oral bacteria could be isolated from atherosclerotic lesions and if DNA from periodontal pathogens could be detected by use of polymerase chain reaction (PCR) techniques. METHODS: Seventy-nine specimens of atherosclerotic plaque removed from carotid or femoral arteries during surgery were immediately transferred to reduced transport fluid and brought to the laboratory. The calcified tissue was meticulously cut into fine pieces and used for cultivation of Porphyromonas gingivalis, Prevotella intermedia, P. nigrescens, Campylobacter rectus, Actinobacillus actinomycetemcomitans, Tannerella forsythensis, and oral streptococci. The material from 24 of the specimens was homogenized, DNA was extracted, and PCR amplification of 16S rDNA with universal and specific primers was carried out. Finally, the PCR products were sequenced. RESULTS: None of the samples yielded growth of the oral bacteria under investigation. In all the 24 specimens bacterial DNA was detected and likewise DNA of P. intermedia was found in the samples. P. nigrescens and P. gingivalis were found sporadically. CONCLUSIONS: Viable oral bacteria could not be isolated from the atheromas, but the data confirm that DNA of periodontal pathogens can be detected in atherosclerotic plaques. However, the finding that DNA from P. intermedia constantly occured in the examined samples was new. Further studies may focus on the simultaneous occurrence of identical clones of this species in subgingival plaque and atherosclerotic plaques. PMID- 15898934 TI - Cigarette smoking and apoptosis. AB - BACKGROUND: Epithelial cell hyperplasia and significant increase in thickness of the overlying orthokeratin layer are characteristic findings noted in the oral cavity of subjects who smoke. Increased proliferation of epithelial cells or defective apoptosis may play a role in the development of epithelial hyperplasia. Thus we analyzed soluble Fas and nuclear matrix protein (NMP) levels in the saliva of smokers (N = 13) and non-smokers (N = 14) to assess apoptosis. METHODS: Ten ml of unstimulated saliva samples was obtained from 14 non-smoker and 13 smoker subjects with the spitting method. These samples were analyzed by using an immunoassay kit to detect soluble human APO-1/Fas and cell death detection enzyme linked immunosorbent assay (ELISA) kit based on nuclear matrix protein 41/7 qualification. RESULTS: The mean soluble Fas levels were 153.8 +/- 290 pg/ml and 315.4 +/- 490 pg/ml and NMP levels were 21.81 +/- 10.70 U/ml and 30.31 +/- 19.86 U/ml, respectively, in smokers and nonsmokers. The difference between NMP levels of smoker and non-smoker groups was statistically significant (P = 0.05). CONCLUSION: The results of the present study suggest that smoking may induce anti apoptotic mechanism in the oral cavity. PMID- 15898935 TI - Characteristics of periodontal microflora in acute myocardial infarction. AB - BACKGROUND: Periodontitis has been linked to increased risk of cardiovascular diseases. Systemic reactions associated with cardiovascular events may depend on characteristics of the subgingival microflora in periodontitis. Our objectives were to compare the numbers of cultivable bacteria, composition of subgingival microflora and clonal distribution of Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) in two groups of patients with generalized chronic periodontitis (GCP), one with an acute myocardial infarction (AMI-GCP) and the other one without AMI (non-AMI-GCP). METHODS: In all, 150 dentate individuals were screened for suitability to this study. Subgingival bacterial samples were collected from 11 AMI-GCP and 11 non-AMI-GCP patients who had been selected using strict inclusion criteria in an attempt to exclude confounding factors and to increase comparability of periodontal conditions by matching for periodontal probing depths and attachment levels. Culture methods were used to determine the total viable counts and occurrence and proportions of six periodontal bacterial species and yeasts. Polymerase chain reaction (PCR) technique was used to detect A. actinomycetemcomitans and Porphyromonas gingivalis (P. gingivalis). Intraspecies characterization of A. actinomycetemcomitans included serotyping and genotyping. RESULTS: The mean proportions of P. gingivalis (P = 0.05) and Tannerella forsythensis (T. forsythensis) (P = 0.01) were significantly lower, but the numbers of Micromonas micros (M. micros) and A. actinomycetemcomitans were up to nine times higher and the mean total number of cultivable bacteria per sample higher (P <0.01) in AMI-GCP than in non-AMI-GCP. CONCLUSION: The findings that no target subgingival species were overrepresented but the total bacterial number was higher in AMI-GCP than non-AMI-GCP patients may provide support to the hypothesis that elevated numbers of bacteria in close vicinity to sterile parenteral area present a risk for systemic health. PMID- 15898936 TI - Adjunctive antimicrobial therapy of periodontitis: long-term effects on disease progression and oral colonization. AB - BACKGROUND: This study assessed the long-term effects of adjunctive antimicrobial therapy on periodontal disease progression and oral colonization. METHODS: Patients with previously untreated chronic periodontitis and subgingival Actinobacillus actinomycetemcomitans and/or Porphyromonas gingivalis were randomly assigned to subgingival scaling without (control group) or with systemic amoxicillin plus metronidazole and CHX irrigation (test group). Relative attachment levels were determined and subgingival and mucosal plaque samples were taken at baseline, at 10 days (plaque only) and at 3, 6, 9, 12, 18, and 24 months following initial therapy. During maintenance therapy, patients received supragingival debridement only. RESULTS: After 24 months, the 18 test group patients showed at sites with initial probing depths (PD) > or =7 mm a significantly (P <0.05) higher frequency of 2 mm or more attachment gain (37.3% +/- 4.6%) and lower frequency of attachment loss (7.2% +/- 3.1%) compared to the 17 controls (8.2% +/- 3.9% and 19.1% +/- 13.1%, respectively). Compared to controls, the intraoral prevalence of A. actinomycetemcomitans (up to 18 months) and P. gingivalis (up to 3 months) decreased and that of Eikenella corrodens (at 10 days) increased in test patients (P <0.05). In both treatment groups, the detection frequency of Tannerella forsythensis decreased transiently, while an overall increase was recorded for Treponema ssp. CONCLUSIONS: Over the 24-month period, a single course of the administered adjunctive antimicrobial therapy led to a relative risk reduction of 62% for attachment loss at deep sites. However, with the exception of A. actinomycetemcomitans, it failed to induce long-term changes in the prevalence profiles of oral colonization. PMID- 15898937 TI - In vitro evidence that the biological effects of platelet-rich plasma on periodontal ligament cells is not mediated solely by constituent transforming growth factor-beta or platelet-derived growth factor. AB - BACKGROUND: The biological actions of platelet-rich plasma (PRP) are thought to be mediated primarily by constituent transforming-growth factor-beta1 (TGF-beta1) and platelet-derived growth factor-AB (PDGF-AB). However, we previously demonstrated that type I collagen expression in periodontal ligament (PDL) cells is acutely stimulated through fibrin clot formation produced by the fibrinogen within PRP, rather than by the known growth factors. To investigate the possible involvement of other unidentified components in PRP action, we have now compared the effects of PRP with those of known recombinant growth factors on cell proliferation, alkaline phosphatase (ALP) activity, and collagen synthesis in human PDL cell cultures. METHODS: PRP was prepared by an established two-step centrifugation protocol using blood obtained from adult human volunteers. Cells cultured in serum-reduced medium on native or collagen-coated plates were treated with PRP, TGF-beta1, or PDGF-AB. Cellular DNA synthesis was evaluated by bromodeoxyuridine incorporation. ALP activity was assessed using p nitrophenylphosphate with formalin-fixed cells, and cellular DNA content was subsequently quantified using bis-benzimide. Collagen synthesis was evaluated using a specific dye-based assay kit. RESULTS: 1) As did both TGF-beta1 and PDGF AB, PRP stimulated cell proliferation. 2) However, only the initial mitogenic action of PRP was attenuated in collagen-coated plates. 3) PRP, but neither growth factor, immediately induced fibrin clot formation and subsequently stimulated cellular adhesion and collagen synthesis. 4) These effects were significantly augmented on collagen-coated plates. 5) PRP enhanced ALP activity, but neither TGF-beta1 nor PDGF-AB replicated this effect. CONCLUSIONS: When evaluated versus the concentrations of growth factors known to be contained by our PRP preparations, these data support the concept that PRP-constituent TGF beta1 acts as a significant growth factor on PDL cells. However, our findings also strongly suggest that the PRP-induced increase in ALP activity is mediated by an as-yet-unidentified component(s). In conjunction with previously demonstrated fibrinogen-mediated actions, our data provide evidence that PRP produces a number of potent effects on PDL cells that does not solely reflect simple combination of its major known growth factors. PMID- 15898938 TI - Evaluation of demineralized bone matrix paste and putty in periodontal intraosseous defects. AB - BACKGROUND: Demineralized bone matrix (DBX) paste and putty are particulate demineralized bone matrices in a 2% or 4% hyaluronate carrier, respectively. The purpose of this study was to determine the effectiveness of DBX paste and putty compared to demineralized freeze-dried bone allograft (DFDBA) in the treatment of human intraosseous periodontal defects. METHODS: Sixty systemically healthy individuals between the ages of 31 and 71 years with at least one intraosseous periodontal defect of > or = 3 mm in depth and radiographic evidence of at least 40% to 50% vertical bone loss were accrued. Following initial non-surgical periodontal therapy, sites were randomly selected to receive either DBX paste, DBX putty, or DFDBA (control). Baseline and 6-month reentry soft and hard tissue parameter measurements were made by calibrated examiners. Data were analyzed within and between groups utilizing analysis of variance (ANOVA) and paired and unpaired Student t tests. RESULTS: Probing depth reductions were significantly improved in all treatment groups with DFDBA, DBX paste, and putty patients demonstrating 2.8 mm, 3.6 mm, and 2.3 mm, respectively. Attachment level gains were significantly improved from baseline for all treatment groups with DFDBA, DBX paste, and putty, respectively, demonstrating 2.4 mm, 2.9 mm, and 1.6 mm. Bone fill was similar between all groups with DBX paste, putty, and DFDBA control groups demonstrating 2.0 mm, 2.4 mm, and 2.2 mm, respectively. All groups yielded significant improvements in percent bone fill with DFDBA, DBX paste and putty, respectively, achieving 37%, 42.1%, and 50% with no significant differences between the groups. CONCLUSION: In summary, demineralized bone matrix paste, demineralized bone matrix putty, and demineralized freeze-dried bone allograft all demonstrated similar favorable improvements in soft and hard tissue parameters in the treatment of human intraosseous defects. PMID- 15898939 TI - Effects of non-surgical mechanical therapy on the subgingival microbiota of Brazilians with untreated chronic periodontitis: 9-month results. AB - BACKGROUND: Mechanical periodontal therapy is the most common treatment of periodontal infections. It is directed primarily towards removing biofilm and calculus from the root surfaces, leading to ecological changes in the subgingival environment. Thus, the purpose of this study was to evaluate the effects of scaling and root planing (SRP) on the subgingival microbiota of Brazilian subjects with untreated chronic periodontitis over a 9-month period. METHODS: Twenty-five untreated chronic periodontitis patients (mean age 43 +/- 5 years; 20% smokers; 45% males) were selected from a Brazilian population. At baseline, probing depth (PD), clinical attachment level (CAL), visible supragingival biofilm (SB), bleeding on probing (BOP), and suppuration (SUP) were measured at six sites/tooth. Subgingival plaque samples were obtained from 10 sites with the deepest PD (> or =5 mm) of each subject and tested for the presence of 25 oral species by DNA probes and the checkerboard technique. Patients received full mouth SRP and oral hygiene instructions. Clinical and microbiological assessments were repeated at 3, 6, and 9 months after therapy. During this period, all patients received maintenance therapy, including supragingival prophylaxis and reinforcement in home care procedures. The clinical and microbiological parameters examined were computed for each subject and at each visit. Differences over time were sought using the Friedman test. RESULTS: Significant reductions in mean CAL and PD (P <0.01), percent of sites with SB (P <0.01), BOP and SUP (P <0.05) were observed during the course of the study. In general, microbial changes were more pronounced for the mean counts than for the frequency of the microorganisms, particularly at 3 months post-therapy. Significant reductions in prevalence and levels were observed for certain periodontal pathogens including P. gingivalis (P <0.05; P <0.01), T. forsythensis (P <0.01), C. rectus (P <0.01), and A. actinomycetemcomitans (P <0.01; P <0.05). Nevertheless, the frequency of A. actinomycetemcomitans increased to baseline values at 9 months after therapy. Treponema ssp. and Prevotella spp. showed a modest decrease in prevalence, whereas marked reductions in their levels were observed. In contrast, the frequency and counts of the suspected pathogens P. micros and F. nucleatum increased after treatment. Species considered beneficial including Actinomyces spp., some oral streptococci, and V. parvula increased in prevalence, although these two last species tended to return to baseline levels at 9 months. CONCLUSION: In Brazilians with untreated chronic periodontitis, SRP led to clinical improvement associated with a decrease of certain periodontal pathogens, and an increase of beneficial species for up to 9 months after therapy. PMID- 15898940 TI - Interleukin-18 concentrations and the pathogenesis of periodontal disease. AB - BACKGROUND: Interleukin-18 (IL-18) is reported as an important regulatory cytokine in non-oral inflammation. Our objective was to compare the concentrations of IL-18 within diseased and healthy human gingiva with concentrations of other T(H)1 and T(H)2 cytokines to determine possible effects of IL-18 on gingival inflammation. METHODS: Gingival biopsies were obtained prior to routine tooth extraction. Gingiva was grouped by the depth of the adjacent gingival sulcus: < or =3 healthy (featuring no bleeding on probing) and > or =3 mm diseased (featuring bleeding on probing). Diseased gingiva was subdivided into 3, 4 to 6 and >6 mm groups. Gingival interleukin (IL)-2, IL-4, IL-6, IL-10, IL 12, IL-18, and interferon (IFN)-gamma concentrations were assessed by enzyme linked immunosorbent assay (ELISA). Data were compared by factorial analysis of variance and the Pearson's correlation test. RESULTS: Concentrations of IL-2, IL 4, IL-6, IL-10, IL-18, and IFN-gamma adjacent to 4 to 6 mm diseased sites were greater than adjacent to < or =3 mm healthy sites (P <0.001). IL-12 concentrations were lower within diseased than within healthy gingiva (P <0.001). IL-6 and IL-18 concentrations were greater adjacent to >6 mm sites compared to healthy sites (P <0.001); the concentrations of the other cytokines (except IL 12) were similar to healthy sites. IL-6 and IL-18 concentrations were positively correlated, and IFN-gamma and IL-12 negatively correlated, with the adjacent gingival sulcular depth. CONCLUSIONS: Periodontal inflammation may not successfully resolve because of accumulation of IL-6 and IL-18, and decreased concentrations of IL-12, within diseased gingiva. Because of the highly significant correlation between IL-18 concentration and gingival sulcular depth, IL-18 may be a useful target for either preventive or palliative therapy for periodontitis. PMID- 15898941 TI - Crestal bone changes around titanium implants. Part I: A retrospective radiographic evaluation in humans comparing two non-submerged implant designs with different machined collar lengths. AB - BACKGROUND: Experimental studies demonstrated that peri-implant crestal hard and soft tissues are significantly influenced in their apico-coronal position by the rough/smooth implant border as well as the microgap/ interface between implant and abutment/restoration. The aim of this study was to evaluate radiographically the crestal bone level changes around two types of implants, one with a 2.8 mm smooth machined coronal length and the other with 1.8 mm collar. METHODS: In 68 patients, a total of 201 non-submerged titanium implants (101 with a 1.8 mm, 100 with a 2.8 mm long smooth coronal collar) were placed with their rough/smooth implant border at the bone crest level. From the day of surgery up until 3 years after implant placement crestal bone levels were analyzed digitally using standardized radiographs. RESULTS: Bone remodeling was most pronounced during the unloaded, initial healing phase and did not significantly differ between the two types of implants over the entire observation period (P >0.20). Crestal bone loss for implants placed in patients with poor oral hygiene was significantly higher than in patients with adequate or good plaque control (P <0.005). Furthermore, a tendency for additional crestal bone loss was detected in the group of patients who had been diagnosed with aggressive periodontitis prior to implant placement (P = 0.058). In both types of implants, sand-blasted, large grit, acid-etched (SLA) surfaced implants tended to have slightly less crestal bone loss compared to titanium plasma-sprayed (TPS) surfaced implants, but the difference was not significant (P >0.30). CONCLUSION: The implant design with the shorter smooth coronal collar had no additional bone loss and may help to reduce the risk of an exposed metal implant margin in areas of esthetic concern. PMID- 15898942 TI - Analysis of interleukin-activated human gingival fibroblasts: modulation of chemokine responses by female hormones. AB - BACKGROUND: The female sex hormones are known to affect the response of numerous tissues to an immune challenge. Because such hormones normally fluctuate during puberty, pregnancy, and the menstrual cycle, more information about the hormonal modulation of such responses in the oral cavity is needed. Gingival fibroblasts (GF), major components of the oral tissues, are potentially sources for inflammatory mediators. METHODS: Macroarrays specific for cytokines and related proteins were used to examine the regulation of gene expression in GF under serum free, resting conditions, after immune challenge with interleukin-1beta (IL 1beta), and in the presence of IL-1beta plus a progestin, +/-17beta-estradiol. Additional studies used enzymelinked immunosorbent assays (ELISAs) to test for secreted chemokines after the same treatments. RESULTS: Of the 392 genes on the macroarray, 66 were up- or downregulated at least 2-fold relative to the unstimulated control in an average of six different sub-lines. Chemokines represented the largest group (18%) of these regulated genes. Numerous genes whose expression was upregulated by IL-1beta were modulated downward by IL-1beta plus progestin, +/-17beta-estradiol. Measurements of the secretion of IL-8, a CXC chemokine, and MCP-1, a CC chemokine, confirmed the inhibitory effect of a progestin on these genes. CONCLUSIONS: Gingival fibroblasts are active participants in the immune response in the oral cavity, and may potentially produce many chemokine signals after exposure to IL-1beta. GF can attract neutrophils, monocytes, eosinophils, and fibroblasts to the area of injury, and aid in the wound repair process. The concentration of female sex hormones, especially progestin, may significantly affect these signaling systems. PMID- 15898943 TI - Prostaglandin E(2) is a main mediator in receptor activator of nuclear factor kappaB ligand-dependent osteoclastogenesis induced by Porphyromonas gingivalis, Treponema denticola, and Treponema socranskii. AB - BACKGROUND: Periodontitis is an inflammatory disease that often leads to destruction of alveolar bone; a number of bacteria in subgingival plaque are associated with bone destruction in periodontitis. To understand the mechanism of how periodontopathogens induce osteoclastogenesis, we determined which mediators are involved in the osteoclastogenesis. METHODS: We investigated effects of sonicates from three periodontopathic bacteria, Porphyromonas gingivalis, Treponema denticola, and Treponema socranskii, on osteoclast formation in a co culture system of mouse calvaria-derived osteoblasts and bone marrow cells. The osteoclast formation was determined by tartrate resistant acid phosphatase (TRAP) staining. The expression of the receptor activator of nuclear factor-kappa B ligand (RANKL), prostaglandin E(2) (PGE(2)) and osteoprotegerin (OPG) in mouse calvaria-derived osteoblasts was determined by immunoassay. RESULTS: Each bacterial sonicate induced the osteoclast formation in the co-culture system. These bacterial sonicates increased the expression of RANKL and PGE(2), and decreased the expression of OPG in osteoblasts. The addition of OPG, an inhibitor of RANKL, in the co-culture completely suppressed the osteoclastogenesis that was stimulated by each bacterial sonicate. Indomethacin, which is an inhibitor of PGE(2) synthesis, reduced more than 88% of the osteoclast formation induced by each bacterial sonicate. Indomethacin inhibited more than 80% of RANKL expression in osteoblasts induced by T. denticola and T. socranskii, and 59% by P. gingivalis. Indomethacin completely recovered the depression of OPG expression in osteoblasts by T. denticola and T. socranskii to the level of the untreated osteoblasts. Indomethacin recovered the reduction of OPG expression by P. gingivalis to 67%. CONCLUSION: These findings suggest that the osteoclastogenesis by P. gingivalis, T. denticola, and T. socranskii is mediated by a RANKL dependent pathway and that PGE(2) is a main factor in the pathway by the enhancing of RANKL expression and the depression of osteoprotegerin, a RANKL inhibitor. PMID- 15898944 TI - Treatment options following single-rooted tooth removal: a literature review and proposed hierarchy of treatment selection. AB - BACKGROUND: Alveolar bone changes following tooth extraction have been well documented and have given rise to a number of treatment approaches. Included in these approaches are placement of various grafting materials, immediate implant placement, and a combination of both. METHODS: A review of all pertinent literature discussing regenerative therapy at the time of tooth extraction or immediate implant placement with or without concomitant regenerative therapy was carried out. RESULTS: A clinically-based hierarchy of treatment selection following extraction of single rooted teeth is proposed, based upon the available literature and clinical experience. The role of patient phenotype is considered. CONCLUSION: Utilization of the proposed hierarchy of treatment selection affords a logical framework within which to predictably treat a variety of patients. PMID- 15898945 TI - Reactive correction of a maxillary incisor in single-tooth crossbite following periodontal therapy. AB - BACKGROUND: The reactive correction of a single tooth anterior crossbite following periodontal therapy is described. This case report provides new information regarding correction of a crossbite relationship and con- firms existing reports of tooth movement following periodontal therapy. METHODS: A 39 year-old woman in good general health presented with a history of recurrent periodontal abscesses of a maxillary incisor. Probing depths of the abscessed tooth ranged from 5 to 12 mm, and class 1 mobility was noted. Radiographs revealed that the tooth had previously been treated endodontically. The patient's periodontal diagnosis was generalized chronic moderate to severe periodontitis. Treatment considerations were complicated by a single-tooth crossbite relationship of the involved incisor and clinical evidence that the periodontal abscess communicated with an apical infection. Treatment of the abscess consisted of cause-related therapy, bone grafting, and occlusal adjustment. RESULTS: Five months after surgical treatment, an edge-to-edge incisal relationship was observed, the first indicator of tooth movement. Further correction to a normal incisal relationship resulted 1 year after modification of the proximal contact. At this time, there was normal probing depth with only slight recession and mobility. Bone fill was radiographically noted. CONCLUSION: It appears that some cases of maxillary incisor crossbite that are complicated by periodontal disease may be corrected, without orthodontic appliances, following periodontal treatment. PMID- 15898946 TI - Periodontal status in two siblings with severe congenital neutropenia: diagnosis and mutational analysis of the cases. AB - BACKGROUND: Severe congenital neutropenia (SCN), also known as Kostmann syndrome, was originally reported as an autosomal recessive disease of neutrophil production. The disease is characterized by a maturation arrest of neutrophil precursors at the promyelocytic stage of differentiation and by extremely low levels of mature neutrophils in peripheral blood. METHODS: A 6-year-old male presented with a complaint of gingival swelling and bleeding, and swelling at the left side of his face. Upon clinical examination, severe inflammation of all gingival tissues was apparent, and a periapical abscess with mobility was noted on the left mandibular second molar. Medical and dental histories revealed numerous recurrent bacterial infections associated with oral and non-oral tissues. His medical history with recurrent infections led us to evaluate his 3 year-old sister to determine the status of her oral health. Inflammation of her oral tissues and recurrent bacterial infections were apparent. Their consanguineous parents were in good health. To assist in identifying possible systemic diseases underlying the inflammatory situation in the siblings, consultations were requested from the Pediatric Hematology Department at Selcuk University and Pediatric Oncology Department at Gulhane Military Medical Academy. RESULTS: Based on absolute neutrophil count (< or =200/mm(3)) and bone marrow aspiration findings consistent with early maturation arrest in myelopoiesis, the cases were diagnosed as SCN. No chromosomal abnormality was detected upon cytogenetic examination. Sequencing analysis also revealed no mutation in the neutrophil elastase or growth factor independent-1 (GFI-1) genes in these patients. Severe periodontal disease, attachment loss, and mobility for over 50% of the deciduous teeth were noted. Within 6 months, the male sibling lost all of his deciduous teeth due to periapical and periodontal infections. His sister presented with tooth mobility for all mandibular incisors. Monthly visits, including scaling, polishing, and 0.2% chlorhexidine digluconate irrigation were performed to support their oral hygiene and to avoid recurrent oral infections. We have been able to stabilize these patients' periodontal conditions during a 2 year follow-up period. CONCLUSION: This case report emphasizes the role of periodontists and pediatric dentists in the diagnosis of diseases linked with neutrophil and other systemic disorders and highlights the need to optimize the health of oral tissues with regular appointments. PMID- 15898947 TI - A new incision design for mandibular symphysis bone-grafting procedures. AB - BACKGROUND: Traditional methods of procuring mandibular symphysis bone grafts may leave soft tissue scarring, and cause paresthesia and lip droop. METHODS: Nineteen patients selected for treatment were given general health, periodontal, and radiographic evaluations. Patients had inadequate bone volume for dental implant placement or required preprosthetic ridge augmentation procedures. Prior to surgery, bone sounding was performed to determine tissue thickness. All patients had a minimum of 4 mm of keratinized gingiva. Under local anesthesia, incisions were initiated within the keratinized gingiva. Full-thickness mucoperiosteal flaps were elevated, and small burs were used to obtain bone blocks from the mandibular symphysis. A bone-scraping device was used to obtain strips of cortical bone. A combination of sling and interrupted sutures was used for wound closure. RESULTS: All patients healed uneventfully without wound dehiscence, paresthesia, or lip droop. Sufficient bone was obtained for ridge or sinus augmentation with eventual implant placement. CONCLUSIONS: A new incision design is presented. This flap design is carried out within keratinized gingiva. Limiting the flap design to keratinized tissue facilitates flap closure and avoids wound dehiscence. PMID- 15898948 TI - Informational paper: implications of genetic technology for the management of periodontal diseases. PMID- 15898949 TI - Collagen reorganization in leech wound healing. AB - BACKGROUND INFORMATION: Leeches respond to surgical lesions with the same sequence of events as that described for wound healing in vertebrates, where collagen is important for the development of tensions in healing wounds, functioning as an extracellular scaffold for accurate regeneration of the structures disrupted by surgical or traumatic actions. RESULTS: In surgically lesioned leeches, newly synthesized collagen is arranged in hierarchical structures. Fibrils can be packed and shaped to form cords or tubular structures, thus acting as an extracellular scaffold that directs and organizes the outgrowth of new vessels and the migration of immune cells towards lesioned tissues. In these animals, the general architecture of collagen fibrils, generated during tissue regeneration, shows similarities to both the structural pattern of collagen bundles and assembly processes observed in several vertebrate systems (fish scales, amphibian skin and human cornea). CONCLUSIONS: The production of extracellular matrix during wound healing in leeches is a surprising example of conservation of an extremely close relationship between the structure and function of molecular structures. It could be hypothesized that collagen structures, characterized not only by a striking structural complexity, but also by multifunctional purposes, are anatomical systems highly conserved throughout evolution. PMID- 15898950 TI - Expression and localization of two isoforms of AQP10 in human small intestine. AB - BACKGROUND INFORMATION: AQP10 (aquaporin 10) is a new member of water channels and two different transcripts have been reported. The difference between these transcripts is in the presence or the absence of an insertion of 475 nt, which leads to a translation frame shift resulting in a different termination of amino acid translation and synthesis of a shorter form of AQP10. Thus an AQP10 molecule deduced from the transcript with an insertion has a distinct C-terminus, different from that translated from the transcript without the insertion. In the present study, we investigated the expression and localization of these AQP10 isoforms. RESULTS: Two transcripts were expressed in human duodenum and jejunum. Western-blot analysis using antibodies for these AQP10 isoforms immunoblotted two bands of approx. 35 and 30 kDa respectively. Immunohistochemistry and immunoelectron microscopy demonstrated localization of the AQP10 with an insertion in the capillary endothelium in villi of the small intestine and the isoform without the insertion in the gastro-entero-pancrestic endocrine cells. CONCLUSION: The different forms of localization of the AQP10 isoforms indicate that they may play different roles in the proximal portion of human small intestine. PMID- 15898951 TI - The role of Munc18-1 in docking and exocytosis of peptide hormone vesicles in the anterior pituitary. AB - BACKGROUND INFORMATION: Many neurons secrete classical transmitters from synaptic vesicles as well as peptide transmitters from LDCVs (large dense-core vesicles). Little is known about the mechanistic differences between these two secretory pathways. The soluble protein Munc18-1 is essential for synaptic vesicle secretion [Verhage, Maia, Plomp, Brussaard, Heeroma, Vermeer, Toonen, Hammer, van den Berg, Missler, et al. (2000) Science 287, 864-869.]. RESULTS: In the present study, we tested if Munc18 genes are also involved in peptidergic secretion from LDCVs using the anterior pituitary as a model system. We show that Munc18-1 is the dominant isoform expressed in the anterior pituitary. In Munc18-1 null mutant mice, the anterior pituitary developed normally and the five major endocrine cell types had a normal distribution. However, circulating peptide hormone levels were decreased by up to 50-fold in the null mutant, whereas the intracellular levels were significantly higher than that in controls. Ultrastructural analysis using the tannic acid method revealed striking differences in the distribution of secretory vesicles: (i) the number of exocytotic figures was mostly decreased in the null mutants and (ii) the LDCVs accumulated near but not at their target membrane. This is in contrast with the apparently normal distribution of synaptic vesicles in developing synapses in the null mutant (Verhage et al., 2000). CONCLUSIONS: We conclude that Munc18-1 is involved in the secretion of peptide hormones and in the docking of LDCVs. These results unmask an apparent mechanistic difference between LDCVs and synaptic vesicles. PMID- 15898952 TI - Centrioles resist forces applied on centrosomes during G2/M transition. AB - BACKGROUND INFORMATION: Centrosome movements at the onset of mitosis result from a balance between the pulling and pushing forces mediated by microtubules. The structural stability of the centrosome core structure, the centriole pair, is correlated with a heavy polyglutamylation of centriole tubulin. RESULTS: Using HeLa cells stably expressing centrin-green fluorescent protein as a centriole marker, we monitored the effect of microinjecting an anti-(polyglutamylated tubulin) monoclonal antibody, GT335, in G1/S or G2 cells. In contrast with the slow effect of the monoclonal antibody GT335 during interphase, a dramatic and rapid centrosome fragmentation occurred in cells microinjected in G2 that was both Eg5- and dynein-dependent. Inhibition of either one of these two motors significantly decreased the scattering of centrosome fragments, and inhibition of centrosome segregation by impairing microtubule dynamics abolished centrosome fragmentation. CONCLUSIONS: Our results demonstrate that the compact structure of the mitotic centrosome is capable of absorbing most of the pulling and pushing forces during G2/M transition and suggest that centrosomes could act as mechanosensors integrating tensions during cell division. PMID- 15898953 TI - Specific plasma membrane aquaporins of the PIP1 subfamily are expressed in sieve elements and guard cells. AB - BACKGROUND INFORMATION: Transmembrane water flow is aided by water-specific channel proteins, aquaporins. Plant genomes code for approx. 35 expressed and functional aquaporin isoforms. Plant aquaporins fall into four different subfamilies of which the PIPs (plasma membrane intrinsic proteins) constitute the largest and evolutionarily most conserved subfamily with 13 members in Arabidopsis and maize. Furthermore, the PIPs can be divided into two phylogenetic groups, PIP1 and PIP2, of which the PIP1 isoforms are most tightly conserved, sharing >90% amino acid sequence identity. As the nomenclature implies, the majority of PIPs have been shown to be localized at the plasma membrane. Recently, two highly abundant plasma membrane aquaporins, SoPIP2;1 and SoPIP1;2, have been purified and structurally characterized. RESULTS: We report the cloning of a cDNA encoding SoPIP1;2 and show that there are at least five additional sequences homologous with SoPIP2;1 and SoPIP1;2 in the spinach genome. To understand their role in planta, we have investigated the cellular localization of the aquaporin homologues SoPIP1;2 and SoPIP1;1. By Western- and Northern-blot analyses and by immunocytochemical detection at the light and electron microscopic levels, we show that SoPIP1;2 is highly expressed in phloem sieve elements of leaves, roots and petioles and that SoPIP1;1 is present in stomatal guard cells. CONCLUSIONS: Localization of the two abundant aquaporin isoforms suggests roles for specific PIPs of the PIP1 subgroup in phloem loading, transport and unloading, and in stomatal movements. PMID- 15898954 TI - Identification of a keratinocarcinoma cell line expressing AQP3. AB - BACKGROUND INFORMATION: AQP3 (aquaporin 3) in the skin is important for skin moisture as demonstrated by the studies of AQP3-null mice, which have accelerated skin drying. Prevention of dry skin is important not only from a cosmetic but also from a clinical point of view. Primary keratinocyte cultures are cumbersome for screening substances that modulate AQP3 expression. RESULTS: A human keratocarcinoma cell line was found to express AQP3 mRNA and protein, which responded to hypertonic stimulation with sorbitol, suggesting that the AQP3 expression is normally regulated in this cell line. This cell line also expressed the type 1 keratinocyte transglutaminase gene. The AQP3 expression was unaffected by all-trans-retinoic acid up to 10(-6) M. Similarly, the retinoic acid did not increase the AQP3 expression up to 1% concentration in rat skin. CONCLUSION: This cell line is useful for the screening of candidate substances that modulate AQP3 expression. PMID- 15898955 TI - Altered expression of aquaporin 4 and H(+)/K(+)-ATPase in the stomachs of peptide YY (PYY) transgenic mice. AB - BACKGROUND INFORMATION: The hormone PYY (peptide YY), synthesized by endocrine cells in the pancreas, ileum, colon and stomach has widespread inhibitory effects on gastrointestinal and pancreatic fluid secretion. Transgenic mice expressing a viral oncoprotein under the control of the PYY gene 5'-flanking region develop well-differentiated colonic endocrine tumours producing mainly PYY and enteroglucagon. In the present study, we investigated the expression of AQP4 (aquaporin 4) water channel and H(+)/K(+)-ATPase in stomachs from both control and transgenic mice. RESULTS: Semi-quantitative RT (reverse transcriptase)-PCR showed an increase in the AQP4 transcript compared with control mice. Quantitative Western-blot analysis of stomachs from control and transgenic mice confirmed a significant increase in the 30 kDa AQP4 protein in transgenic mice. In control mice, AQP4 is specifically expressed in the basolateral membrane of gastric parietal cells, located in the basal region of the fundic glands. This particular location suggests that parietal cells in the base region of gastric pits might have a major role in water transport when compared with the more superficial parietal cells. Interestingly, immunofluorescence studies on transgenic mice revealed that the quantitative increase of AQP4 expression was actually due to an increase in the number of AQP4-expressing epithelial cells rather than to a higher expression of AQP4 in parietal cells. In fact, immunofluorescence experiments using the specific antibody raised against the AE2 isoform of Cl(-)/HCO3- exchanger specifically expressed in parietal cells confirmed that the number of parietal cells was comparable in both PYY and control stomachs. Moreover, in transgenic mice, a parallel significant decrease in the expression of H(+)/K(+)-ATPase was observed, as revealed by RT-PCR, quantitative immunoblotting and immunofluorescence. CONCLUSIONS: In the present study, we demonstrate that the sustained inhibition of gastric secretion due to tumours producing PYY/enteroglucagon in transgenic mice is associated with an increase in AQP4 expression and a down-regulation of H(+)/K(+)-ATPase in parietal cells that acquire the characteristics of basal parietal cells. The absence of H2 receptors-mediated signalling due to the inhibition of histamine release from ECL (enterochromaffin-like) cells by PYY may be in part responsible for the observed increase in the number of parietal cells expressing AQP4. PMID- 15898956 TI - Decreased expression of AQP2 and AQP4 water channels and Na,K-ATPase in kidney collecting duct in AQP3 null mice. AB - BACKGROUND INFORMATION: Phenotype analysis has demonstrated that AQP3 (aquaporin 3) null mice are polyuric and manifest a urinary concentration defect. In the present study, we report that deletion of AQP3 is also associated with an increased urinary sodium excretion. To investigate further the mechanism of the decreased urinary concentration and significant natriuresis, we examined the segmental and subcellular localization of collecting duct AQPs [AQP2, p-AQP2 (phosphorylated AQP2), AQP3 and AQP4], ENaC (epithelial sodium channel) subunits and Na,K-ATPase by immunoperoxidase and immunofluorescence microscopy in AQP3 null (-/-), heterozygous (+/-) mice, wild-type and unrelated strain of normal mice. RESULTS: The present study confirms that AQP3 null mice exhibit severe polyuria and polydipsia and demonstrated that they exhibit increased urinary sodium excretion. In AQP3 null mice, there is a marked down-regulation of AQP2 and p-AQP2 both in CNT (connecting tubule) and CCD (cortical collecting duct). Moreover, AQP4 is virtually absent from CNT and CCD in AQP3 null mice. Basolateral AQP2 was virtually absent from AQP3 null mice and normal mice in contrast with rat. Thus the above results demonstrate that no basolateral AQPs are expressed in CNT and CCD of AQP3 null mice. However, in the medullary collecting ducts, there is no difference in the expression levels and subcellular localization of AQP2, p-AQP2 and AQP4 between AQP3 +/- and AQP3 null mice. Moreover, a striking decrease in the immunolabelling of the alpha1 subunit of Na,K-ATPase was observed in CCD in AQP3 null mice, whereas a medullary-collecting duct exhibited normal labelling. Immunolabelling of all the ENaC subunits in the collecting duct was comparable between the two groups. CONCLUSIONS: The results improve the possibility that the severe urinary concentrating defect in AQP3 null mice may in part be caused by the decreased expression of AQP2, p-AQP2 and AQP4 in CNT and CCD, whereas the increased urinary sodium excretion may in part be accounted for by Na,K-ATPase in CCD in AQP3 null mice. PMID- 15898957 TI - Inhibition of prostaglandin synthesis does not alter the decrease in pre capillary resistance in the human calf in response to small cumulative increases in venous congestion. AB - The decrease in pre-capillary resistance in the human calf during gradual cumulative increases in venous congestion pressure has been proposed to represent vasodilator signalling between the venous and arterial microcirculations. The present study investigated whether prostaglandins are involved in this local flow regulation by measuring calf blood flow and microvascular filtration capacity using strain gauge plethysmography in young male subjects before (baseline) and after taking either ibuprofen, an inhibitor of prostaglandin synthesis (1600 mg over 2 days), or placebo. At baseline, inflation of a thigh cuff to 50 mmHg in steps of 10 mmHg, each held for 5 min, did not decrease arterial inflow, confirming a reduction of pre-capillary resistance. Ibuprofen reduced resting calf blood flow by 35% (P<0.001), but flow at a Pcuff (cuff pressure) of 50 mmHg was 97% of this value, i.e. pre-capillary resistance had decreased to the same extent as before inhibition of prostaglandin synthesis. Ibuprofen also reduced microvascular filtration capacity (2.98+/-1.20 compared with 3.71+/-0.89 ml.min 1.100 ml-1.mmHg-1x10(-3); P<0.05), probably due to a combination of reduced arterial inflow and lower venous pressure (8.5+/-5.2 compared with 12.6+/-2.8 mmHg; P<0.05) that moderated capillary hydrostatic pressure to override direct effects of inhibition of prostaglandin synthesis on permeability. Placebo was without effect on any measurement. It is unlikely therefore that prostaglandin mediated vasodilator signals, which have been demonstrated between paired veins and arteries, are important in local vasodilation in response to venous congestion. PMID- 15898958 TI - Modulation of lipid metabolism by n-3 polyunsaturated fatty acids in gestational diabetic rats and their macrosomic offspring. AB - The time course of changes in lipid metabolism by dietary n-3 PUFAs (polyunsaturated fatty acids) in streptozotocin-induced diabetic rats during pregnancy (days 12 and 21) and their macrosomic offspring at birth (day 0) and through adulthood (days 60 and 90) was studied with respect to adipose tissue, liver and serum lipid concentrations, and fatty acid composition. Glucose and insulin levels were also assessed in order to characterize the diabetic state of macrosomic offspring. Pregnant diabetic and control rats were fed either an Isio 4 or EPAX diet (enriched with n-3 PUFA). The same diets were also consumed by pups at weaning. Compared with control rats, during pregnancy diabetic rats had a significant elevation in liver and serum triacylglycerol (triglyceride) and cholesterol concentrations. At birth, macrosomic pups had higher serum insulin and glucose levels than control pups. The macrosomic rats maintained accelerated postnatal growth combined with high adipose tissue weight and lipid content through the first 12 weeks of age. The macrosomic pups from diabetic rats fed the Isio-4 diet also showed a significant enhancement in liver and serum triacylglycerol and cholesterol levels at birth and during adulthood. Feeding the EPAX diet to diabetic mothers as well as their macrosomic pups increased serum and liver levels of EPA (eicospentaenoic acid) and DHA (docosahexaenoic acid) with a reduction in arachidonic acid. The EPAX diet induced a significant decrease in liver and serum triacylglycerol and cholesterol concentrations in mothers during pregnancy and in their macrosomic pups during adulthood. Since the EPAX diet improves lipid anomalies considerably in diabetic mothers and their macrosomic offspring, it may prevent long-term metabolic abnormalities associated with macrosomia. PMID- 15898959 TI - Pharmacotherapy of bipolar mixed states. AB - OBJECTIVE: Mixed episodes comprise up to 40% of acute bipolar admissions. They are difficult-to-treat, complex clinical pictures. This review provides an overview of the available literature on the pharmacotherapy of manic-depressive mixed states and suggests treatment options. METHOD: Literature was identified by searches in Medline, Embase and the Cochrane Controlled Trials Register. Studies were considered relevant if they contained the keywords mixed mania, mixed state(s), mixed episode(s), treatment, therapy, study or trial. RESULTS: Overall, there were very few double-blind, placebo-controlled studies specifically designed to treat manic-depressive mixed states. Rather, patients with mixed states comprised a sub-group of the examined patient cohorts. Nevertheless, the data show that acute mixed states do not respond favourably to lithium. Instead, valproate and olanzapine are drugs of first choice. Carbamazepine may play a role in the prevention of mixed states. Antidepressants should be avoided, because they may worsen intraepisodic mood lability. Lamotrigine may be useful in treating mixed states with predominantly depressive symptoms. CONCLUSIONS: More treatment studies specifically designed to treat the complex clinical picture of mixed states are clearly needed. Current treatment recommendations for clinical practice based on the available literature can only target select aspects of these episodes. PMID- 15898960 TI - Neuropsychological dysfunction in bipolar affective disorder: a critical opinion. AB - Data from the imaging literature have led to suggestions that permanent structural brain changes may be associated with bipolar disorder. Individuals diagnosed with bipolar disorder display deficits on a range of neuropsychological tasks in both the acute and euthymic phases of illness, and correlations between experienced number of affective episodes and task performance are commonly reported. These findings have renewed interest in the neuropsychological profile of individuals with bipolar disorder, with deficits of attention, learning and memory, and executive function, asserted to be present. This paper critically reviews five different potential causes of neurocognitive dysfunction in bipolar disorder: (i) iatrogenic, (ii) acute functional changes associated with depression or mania, (iii) permanent structural lesions of a neurodegenerative origin, (iv) permanent structural lesions that are neurodevelopmental in origin, and (v) permanent functional changes that are most likely genetic in origin. Although the potential cognitive effects of residual symptomatology and long-term medication use cannot be entirely excluded, we conclude that functional changes associated with genetically driven population variation in critical neural networks underpin both the neurocognitive and affective symptoms of bipolar disorder. The philosophical implications of this conclusion for neuropsychology are briefly discussed. PMID- 15898961 TI - Pathways to care for patients with bipolar disorder. AB - Bipolar disorder is a chronic, debilitating psychiatric illness with serious ramifications for patients, their families, and society. Despite the availability of effective treatments, this disease often goes untreated due to medical, financial, legal/governmental, and cultural barriers. In this review we explore possible reasons for this problem. Misdiagnosis of bipolar disorders is a common medical barrier. One pathway to care for individuals with bipolar disorder is through referral from primary care, but primary care physicians generally have not received special training in the recognition and management of bipolar disorder. This often leads to diagnostic delays or errors, which prevents timely 'filtering' of patients into specialized care. Using data bases we explored these pathways. Legislation in the USA, such as the Emergency Medical Treatment and Active Labor Act (EMTALA), designed to ensure access to inpatient mental health care, has instead given hospitals financial incentives to limit inpatient mental health care capacities. Reimbursement of mental health care expenses is a significant issue impacting a patient's ability to gain access to care, as bipolar disorder is a costly disease to treat. Improving access to care among the bipolar community will require multilateral strategies to influence the actions and attitudes of patients, communities, providers, health care systems, and state/national governments. In other cultures, barriers to care differ according to a number of factors such as type of services, explanatory models of illness, misdiagnosis and perceptions of care givers. It is essential that clinicians are aware of pathways and barriers so that appropriate and accessible care can be provided. PMID- 15898962 TI - Reproductive function and risk for PCOS in women treated for bipolar disorder. AB - INTRODUCTION: This study examined the reproductive function and prevalence of polycystic ovary syndrome (PCOS) in women with bipolar disorder taking antimanic medications. METHOD: Women aged 18-45 treated for bipolar disorder and not taking steroid contraceptives were recruited to complete questionnaires about their menstrual cycle and to provide blood samples for measurement of a range of reproductive endocrine and metabolic hormone levels. Eighty women participated in completing the questionnaires and 72 of them provided blood samples. RESULTS: Fifty-two of the 80 women (65%) reported current menstrual abnormalities, 40 of which (50%) reported one or more menstrual abnormalities that preceded the diagnosis of bipolar disorder. Fifteen women (38%) reported developing menstrual abnormalities since treatment for bipolar disorder, 14 of which developed abnormalities since treatment with valproate (p = 0.04). Of the 15 patients reporting menstrual abnormalities since starting medication, 12 (80%) reported changes in menstrual flow (heavy or prolonged bleeding) and five (33%) reported changes in cycle frequency. No significant differences were observed between women receiving or not receiving valproate in mean levels of free or total serum testosterone levels. This was true for the total sample and for the sub-group without preexisting menstrual problems. However, within the valproate group, duration of use was significantly correlated with free testosterone levels (r = 0.33, p = 0.02). Three of the 50 women (6%) taking VPA, and 0% of the 22 taking other antimanic medications, met criteria for PCOS (p = 0.20). Other reproductive and metabolic values outside the normal range across treatment groups included elevated 17 alpha-OH progesterone levels, luteinizing hormone: follicle stimulating hormone ratios, homeostatic model assessment (HOMA) values, and low estrogen and dehydroepiandrosterone sulfate (DHEAS) levels. Preexisting menstrual abnormalities predicted higher levels of 17 alpha-OH progesterone, free testosterone, and estrone as well as development of new menstrual abnormalities. Body mass index (BMI) was significantly positively correlated with free testosterone levels and insulin resistance (HOMA) across all subjects, regardless of medication used. CONCLUSIONS: Rates of menstrual disturbances are high in women with bipolar disorder and, in many cases, precede the diagnosis and treatment for the disorder. Treatment with valproate additionally contributes significantly to the development of menstrual abnormalities and an increase in testosterone levels over time. A number of bipolar women, regardless of type of medication treatment received, have reproductive and metabolic hormonal abnormalities, yet the etiology of such abnormalities requires further study. Women with preexisting menstrual abnormalities may represent a group at risk for development of reproductive dysfunction while being treated for bipolar disorder. PMID- 15898963 TI - GSK-3 activity in neocortical cells is inhibited by lithium but not carbamazepine or valproic acid. AB - OBJECTIVES: Lithium (Li(+)) has been suggested to target the enzyme glycogen synthase kinase 3 (GSK-3) as a mechanism of mood stabilization. Inhibition of GSK 3 by a second mood-stabilizer, valproic acid (VPA), has also been reported, but this effect is dependent on cell type. It is currently unknown if carbamazepine (CBZ) inhibits GSK-3 activity. We have sought to compare the inhibitory effect of Li(+), VPA and CBZ on GSK-3 activity. METHODS: We treated rat primary cultured neurones at three times therapeutic drug concentration with CBZ, VPA and Li(+) and examined changes in GSK-3 protein levels, activity and phosphorylation of downstream targets. To eliminate a possible direct effect of these drugs at higher concentrations, we also looked for direct inhibition of both GSK-3 isoforms at a range of concentrations. RESULTS: CBZ, VPA and Li(+) did not change the levels of the GSK-3 or produce an irreversible in vivo effect on GSK-3 activity. Only Li(+) inhibited the phosphorylation of a cytoskeletal target of GSK-3, tau, whereas CBZ and VPA did not. Surprisingly, none of these drugs altered beta-catenin levels in these cells, a process attenuated by GSK-3 activity. Finally, only Li(+) directly inhibits GSK-3 activity (both alpha and beta isoforms) at therapeutic levels in direct biochemical assays. CONCLUSION: Thus we show that neither GSK-3 nor the altered GSK-3 signalling pathway can provide a common mechanism of action of mood-stabilizing drugs in the mammalian brain. PMID- 15898964 TI - Divalproex sodium for pediatric mixed mania: a 6-month prospective trial. AB - OBJECTIVE: This prospective 6-month open trial examined the effectiveness and safety of divalproex sodium (DVPX) in pediatric mixed mania. METHOD: Thirty-four subjects with a mean age of 12.3 (SD = 3.7) years, DSM-IV diagnosis of a current mixed episode and a baseline Young Mania Rating Scale (YMRS) score >20 were treated with DVPX monotherapy. The primary outcome measures were the YMRS and the Child Depression Rating Scale-Revised. Secondary measures were the Clinical Global Impression Scale for Bipolar Disorder (CGI-BP) and the Children's Global Assessment of Functioning Scale (C-GAS). Measures of safety and tolerability were also administered. RESULTS: Effect size (Cohen's d) based on change scores from baseline was 2.9 for the YMRS and 1.23 for the CDRS-R. Response rate (> or =50% change from baseline YMRS score and < or =40 score on CDRS-R at the end of study) was 73.5%. The remission rate (> or =50% change from baseline on YMRS, < or =40 on CDRS-R, CGI-BP-Improvement subscale of < or =2, and > or =51 CGAS score) was 52.9%. Significant improvements (p < 0.001) from baseline were seen for mean scores on all outcome measures (i.e., YMRS, CGI-BP, CDRS-R, and C-GAS). DVPX was safe and well tolerated with no serious adverse events during the 6-month trial. CONCLUSION: This study provides evidence for the effectiveness and safety of DVPX in the treatment of pediatric mixed mania over a 6-month period. Placebo controlled, randomized trials involving larger samples will ultimately shed light on the efficacy of this agent. PMID- 15898965 TI - Drug-drug interactions as a determinant of elevated lithium serum levels in daily clinical practice. AB - OBJECTIVE: Lithium is a drug with a narrow therapeutic window. Concomitantly used medication is a potentially influencing factor of lithium serum concentrations. We conducted a multicentre retrospective case-control study with the aim of investigating lithium-related drug interactions as determinants of elevated lithium serum levels in daily clinical practice. METHODS: Cases were patients with an increase of at least 50% in lithium serum concentrations resulting in an elevated lithium serum level of at least 1.3 mmol/L, and who were not suspected of a suicide attempt. Controls were patients who showed stable lithium serum levels within the therapeutic range. Use and start of non-steroidal anti inflammatory drugs, diuretics, renin-angiotensin inhibitors, theophyllin and antibiotics were investigated as potential determinants of the elevated lithium serum levels. Irregularity in lithium dispensing pattern, change in lithium dosing regimen, age, gender, prescribing physician and laboratory parameters were investigated as potential confounders. RESULTS: We included 51 cases and 51 controls in our study. Five (9.8%) controls and 15 (29.4%) cases used potentially interacting co-medication [OR of 3.83 (95%CI 1.28-11.48)]. Start of potentially interacting co-medication was observed in eight (15.7%) cases and in zero (0%) controls resulting in an OR of 20.13 (95% CI 1.13-359). After adjustment for co medication, irregularity in lithium dispensing pattern, change in lithium dosing regimen, and age, the statistically significant association was lost. We report an OR of 2.70 (95% CI 0.78-9.31) for use of concomitant medication, with a large contribution of antibiotic agents, and an OR of 3.14 (95% CI 1.15-8.61) for irregularity in lithium dispensing pattern. CONCLUSION: Use of co-medication, especially antibiotics, tends to be associated with elevated lithium serum levels. PMID- 15898966 TI - Functional impairment in the remission phase of bipolar disorder. AB - OBJECTIVES: The purpose of this study was to evaluate functional impairment in a group of patients with bipolar disorder in remission and to determine the extent of relationships between overall functioning and current depressive, manic and panic spectrum symptoms. METHOD: A subset of the patient population at the Pittsburgh site of the Systematic Treatment Enhancement Program in Bipolar Disorder (STEP-BD) study was evaluated in this study. The subsample comprises 103 male and female subjects with bipolar I disorder (n = 70), bipolar II disorder (n = 24), schizoaffective disorder - bipolar type (n = 4), or bipolar disorder NOS (n = 5). Subjects were evaluated in a period of remission (at least 4 weeks with no more than two depressive or manic symptoms). Subjects were assessed for overall functional status using the Work and Social Adjustment Scale (WSAS) and for current bipolar and panic spectrum symptoms using the Mood Spectrum Self Report questionnaire (MOODS-SR) and Panic-Agoraphobic Spectrum Self-Report questionnaire (PAS-SR). RESULTS: The median WSAS total score in these remitted subjects was 14, indicating significant functional impairment. Regressing WSAS on current depressive, manic, and panic spectrum total scores, we observed a highly significant depressive spectrum effect (t = 4.9, df = 94, p < 0.0001), but non significant panic and manic spectrum effects (t = 1.3, df = 94, p = 0.19 and t = 1.8, df = 94, p = 0.07, respectively). CONCLUSION: Bipolar disorder is associated with functional deficits even during periods of sustained and substantial remission. The degree of functional impairment is correlated with the degree of depressive spectrum symptoms. PMID- 15898967 TI - Clinical correlates of current level of functioning in primary care-treated bipolar patients. AB - OBJECTIVES: In this study we examined general assessment of functioning (GAF), and its relation to clinical and demographic factors in bipolar patients. A number of studies, mostly from specialized programs, show that bipolar disorder often leads to occupational and social impairment. Here we report data from patients in a primary care setting. METHODS: A total of 252 patients from the Maritime Bipolar Registry with DSM-IV diagnoses of bipolar I or bipolar II disorder participated in the study. GAF ratings during maintenance treatment were compared across clinical and demographic variables. RESULTS: The mean GAF score in this sample was 67 +/- 17 (range 10-100). The GAF scores followed bimodal distribution with mean values of 50.5 +/- 10.3 and 79.0 +/- 10.3. Decreased functioning was found in patients with chronic illness course, history of rapid cycling, suicidal behaviour, psychiatric comorbidity, hypothyroidism, and diabetes mellitus, regardless of treatment of these conditions. There were no differences in the level of functioning between men and women, bipolar I and II patients, those with and without psychotic episodes, hypertension, treatment with antidepressants or antipsychotics. CONCLUSIONS: Functioning in primary care treated bipolar patients in maintenance phase of treatment is decreased not only due to specific disorder-related variables, but also due to frequent comorbidity with other psychiatric and medical conditions. PMID- 15898968 TI - Hashimoto's encephalopathy presenting with bipolar affective disorder. AB - OBJECTIVE: We report on a rare case of Hashimoto's encephalopathy associated with a bipolar affective disorder. CASE REPORT: A 32-year-old woman presented with a bipolar affective disorder and Hashimoto's thyroiditis. Neurological investigations (magnetic resonance imaging of the brain and cerebrospinal fluid) did not reveal any pathological findings except for a pathological electroencephalogram (EEG). Despite consequent antidepressant treatment, the patient remitted only in conjunction with normalization of the EEG after short term treatment with high doses of prednisolone. CONCLUSION: In treatment resistant courses of disorders, the thorough clinical and laboratory investigation of our patient revealed a very efficient treatment strategy. Cases of Hashimoto's encephalopathy associated with the occurrence of a manic episode and hence with bipolar disorder are rare; this is the first reported case. However, clinicians should be alert to this possibility. PMID- 15898969 TI - Bipolar affective disorder in a male with a deletion of Y chromosome -- a case report. AB - We report on a 25-year-old male with bipolar disorder, dysmorphic features and a deletion of the long arm of Y chromosome. A potential association between sex chromosome abnormalities and a susceptibility to major psychiatric disorders has been documented. However there have been very few reports on the coincidence of Y chromosome aberrations with bipolar disorder. Cytogenetic studies have contributed to the identification of several disease genes. Karyotyping of patients with bipolar disorder in order to identify candidate regions for linkage studies has been recommended. PMID- 15898970 TI - Gabapentin withdrawal syndrome in the presence of a taper. AB - OBJECTIVE: To report a case report of a geriatric patient with a 5-year history of gabapentin use for enhanced bipolar control, who was tapered off of gabapentin over 1 week. The patient displayed unique withdrawal symptoms after the taper of gabapentin. METHODS: The patient is an 81-year-old white female with a life-long history of schizoaffective disorder with bipolar type I tendencies who had been prescribed gabapentin for 5 years. RESULTS: The patient displayed moderate upper respiratory tract infection symptoms and somatic complaints 1 day after termination of gabapentin. These symptoms gradually worsened until 10 days after, at which time she acutely developed severe mental status changes, severe somatic chest pain, and hypertension. Physical examination, electrolytes, electrocardiogram, computerized tomography, magnetic resonance imaging, and magnetic resonance angiography were all normal. Upon reintroduction of gabapentin, the patient returned to baseline within 1-2 days. CONCLUSIONS: Gabapentin is widely utilized currently for the chronic treatment of recalcitrant migraines, bipolar illness, pain, and epilepsy. It has a wide therapeutic index with few side effects and drug interactions, is not hepatically metabolized, and is excreted by the kidneys. Past reports have suggested that some withdrawal symptoms can present after 1-2 days upon abrupt discontinuation of gabapentin after chronic use within young to middle-aged patients. These symptoms mimic that of alcohol and benzodiazepine withdrawal purportedly due to a similar mechanism of action. Unique to this case is that this geriatric patient developed debilitating withdrawal symptoms after a gradual, week-long taper of gabapentin along with flu-like symptoms. It is proposed herein that a gabapentin taper should follow a course similar to that of a benzodiazepine taper -- slowly and over a period of weeks to months. PMID- 15898971 TI - A psychotic episode associated with the Atkins diet in a patient with bipolar disorder. PMID- 15898972 TI - Calcineurin inhibitors for the treatment of skin disease: how do they work? PMID- 15898973 TI - Allergen immunotherapy in early childhood: between Scylla and Charybdis! PMID- 15898974 TI - Effects of calcineurin inhibitors on an in vitro assay for chronic urticaria. AB - BACKGROUND: Chronic urticaria is a common skin disorder, which causes considerable morbidity. In approximately 40% of cases, patients have an autoimmune disorder in which functional antibodies cause degranulation of mast cells and basophils, and C5a complement augments this in varying amounts from patient to patient. Since the calcineurin inhibitor ciclosporin has been used in chronic autoimmune urticaria, we examined the effect of ciclosporin and other drugs on the release of histamine from basophils when stimulated by sera from patients with chronic autoimmune urticaria. METHODS: Leucocytes from healthy donors were isolated and incubated in varying concentrations of ciclosporin, ascomycin, methotrexate, diphenhydramine or hydroxyzine for 30 min prior to stimulation with serum from urticaria patients known to have functional immunoglobulin (Ig)G antibodies directed against the alpha subunit of the IgE receptor. Histamine release was then measured. RESULTS: Pre-incubating cells with ciclosporin and ascomycin produced dose-dependent inhibition of histamine release when cells were stimulated by sera of urticaria patients, by purified IgG from these sera, but not by C5a. Inhibition was not prevented by C5a receptor blocking antibodies. No inhibition was seen with methotrexate, diphenhydramine or hydroxyzine. CONCLUSIONS: This is the first demonstration of inhibition of histamine release by calcineurin inhibitors employing sera of patients with chronic autoimmune urticaria. These drugs may work by interfering with intracellular signalling in cells following cross-linking of the IgE receptor, but not following stimulation of the C5a receptor. PMID- 15898975 TI - Post-marketing survey on the safety of sublingual immunotherapy in children below the age of 5 years. AB - BACKGROUND: The age below 5 years is considered a prudential limit for immunotherapy in view of the possible severity of side-effects. Sublingual immunotherapy (SLIT) seems to be safe, but no study in very young children is available. We performed a safety post-marketing surveillance study in children below 5 years. METHODS: Children aged 3-5 years with respiratory allergy receiving SLIT were followed-up for at least 2 years. A diary card for side effects was filled by parents at each dose given. Local and systemic side-effects were graded as: mild (no intervention, no dose adjustment), moderate (medical treatment and/or dose reduction), severe (life threatening/hospitalization/emergency care). The comparative safety of different allergens and regimens was also assessed. RESULTS: One hundred and twenty-six children (mean age 4.2 years, 67 male) were included. Seventy-six (60%) had rhinitis with asthma, 34 (27%) rhinitis only and 16 (13%) only asthma. Immunotherapy was prescribed for mites (62%), grasses (22.2%), Parietaria (11.9%), Alternaria (2.4%) and olive (1.5%). Eighteen children underwent an accelerated build-up. The total number of doses was about 39,000. Nine side effects were reported in seven children (5.6% patients and 0.2/1000 doses). Two episodes of oral itching and one of abdominal pain were mild. Six gastrointestinal side-effects were controlled by reducing the dose. All side effects occurred during up-dosing phase. No difference in terms of safety among the allergens used was observed. CONCLUSION: SLIT is safe also in children under the age of 5 years. PMID- 15898976 TI - The safety of sublingual-swallow immunotherapy: an analysis of published studies. AB - BACKGROUND: As the main target of sublingual immunotherapy (SLIT) is to reduce at most the occurrence of adverse events (AE), safety represents a critical issue. This aspect deserves particular mention when a higher dose of allergen extract than traditional subcutaneous immunotherapy (SCIT) is required to be effective: that may be up to 500 times that employed for SCIT. OBJECTIVE: All published controlled studies concerning SLIT-swallow were analysed to evaluate AE rates. METHODS: Studies were subdivided in two groups: (i) studies using low allergen dose (LAD), i.e. ranging from 1 to 50 times the dose commonly administered with SCIT, and (ii) studies with high allergen dose (HAD), i.e. ranging from 50 to 500 times the dose administered with SCIT. RESULTS: Twenty-five studies were altogether analysed: 13 studies belonged to the low-dose group, 12 belonged to the high-dose group. We considered all patients with at least one AE. Local reactions were significantly more frequent in the LAD group than in the HAD group (P<0.0001), while there was no difference in the rate of systemic reactions. Severe systemic reactions were never reported. CONCLUSION: This study represents the first analysis of the safety of SLIT concerning the allergen dose employed in the treatment. There is evidence that AE occurrence is substantially not dose dependent. This fact highlights two main clinical aspects: the elevated tolerability of SLIT in general and the safety of HAD regimen. PMID- 15898977 TI - Immunotherapy with depigmented glutaraldehyde-polymerized extracts: changes in quality of life. AB - BACKGROUND: Immunotherapy (IT) with modified allergens reduces allergic rhinitis (AR) symptoms and medications requirements. Improvement of quality of life (QOL) is a key point in the treatment of AR. The aim of this study was to provide evidence of changes related to the patient's QOL (well-being) induced by a modified (depigmented glutaraldehyde-polymerized) therapeutic vaccine and of its safety. MATERIAL AND METHODS: Fifty-three patients with a well-documented clinical history of seasonal AR sensitized to Dactylis glomerata and Olea europaea pollens were included in a randomized clinical trial. Twenty-five patients (Group-A) received a mixture of D. glomerata and O. europaea pollen extracts and 28 patients received placebo (group-C). Any adverse event was recorded and graded in accordance with EAACI guidelines. RQLQ was recorded before the treatment (pollen season 2000) and after 1 year of treatment (pollen season 2001). Dose-response skin prick test with each allergen extract was conducted at baseline and at the end of the study. RESULTS: Each patient received 17 injections during this period. All patients completed the trial and no systemic adverse reactions were recorded. Symptom scores (P<0.001) and medication requirements (P<0.001) were significantly reduced in the IT group during the pollen season. This patient group also experienced greater and statistically significant improvement in overall RQLQ score and in five of the seven domains, all of them surpassing the threshold of 'minimal important difference' of 0.5 points. CONCLUSIONS: Results of this study provided evidence that IT with depigmented, glutaraldehyde-modified allergen extracts was well-tolerated and added beneficial effects to AR treatment in pollen allergic patients eliciting an improvement in QOL enough to justify a change in the patient's treatment. PMID- 15898978 TI - Methotrexate therapy of oral corticosteroid-dependent asthmatics reduces serum immunoglobulins: correlation with clinical response to therapy. AB - BACKGROUND: Concomitant methotrexate (MTX) therapy of oral corticosteroid (CS) dependent asthmatics has been shown to spare CS therapy, but the mechanism is unknown. In a previous report, we showed that MTX increases T cell inhibition by CS. In this report we focus on effects of MTX on immunoglobulin concentrations and their possible clinical relevance. OBJECTIVE: To monitor changes in circulating leucocytes and Ig in a group of these patients during MTX therapy, and to relate these changes to clinical 'response' as defined by oral CS reduction. METHODS: Sixteen severe asthmatics dependent on oral prednisolone 15 (7.5-25) mg/day in addition to high dose inhaled CS were treated with MTX 15 mg intramuscularly, weekly for 28 weeks. Prednisolone dosages were maintained constant for 12 weeks then reduced systematically over the next 16 weeks provided that asthma control did not deteriorate. Patients were classified a priori as 'responders' or 'non-responders' to MTX (reduction of initial oral prednisolone requirement by >or=50% or <50%, respectively). Patients were followed-up for a further 12 weeks after MTX withdrawal. Serum Ig and differential blood leucocyte counts were measured at baseline, 12, 28 and 40 weeks. RESULTS: MTX therapy allowed significant, but individually variable, reductions in oral prednisolone dosages (P<0.00001) without alteration of lung function or symptoms. This was associated with significant reductions in mean serum concentrations of Ig of all classes, which reversed following MTX withdrawal. Reductions in IgE and IgG were significantly greater in the MTX 'responders' as compared with 'non-responders', and changes in IgE, IgG and IgM correlated with changes in prednisolone requirements. Differential blood leucocyte counts showed no significant variation. CONCLUSION: MTX therapy reduced oral CS requirements in these severe asthmatics to a degree which correlated with reduced circulating Ig but not lymphopaenia, suggesting a possible cause and effect relationship. These reductions might also contribute to the documented incidence of opportunistic infection in these circumstances. PMID- 15898979 TI - Association of thromboxane A2 receptor gene polymorphism with the phenotype of acetyl salicylic acid-intolerant asthma. AB - BACKGROUND AND OBJECTIVE: The thromboxane A2 receptor (TBXA2R) is a receptor for a potent bronchoconstrictor, TBXA2 which is known to be related to bronchial asthma and myocardial infarction. TBXA2R antagonist and TBX synthase inhibitors have been found to be effective in the management of asthmatic patients. This study was aimed to evaluate whether genetic variants of TBXA2R may be related with development of acetyl salicylic acid (ASA)-intolerant asthma (AIA). METHODS: TBXA2R gene polymorphisms (TBXA2R+795T>C, TBXA2R+924T>C) were determined using a single-base extension method in 93 AIA patients compared with 172 patients with ASA-tolerant asthma (ATA) and 118 normal controls (NCs) recruited from the Korean population. HLA DPB1*0301 genotype was performed using a direct sequencing method. RESULTS: The rare C allele frequency of TBXA2R+795T>C was significantly higher in AIA than in ATA (P=0.03) and the TBXA2R+795T>C polymorphism was also associated with extent of percent fall in forced expiratory volume in 1 s (FEV1) after the inhalation of lysine-acetyl salicylic acid in AIA patients (P=0.009); AIA patients homozygous for the +795 C allele had a greater percent fall of FEV1 compared with individuals with TBXA2R+795 CT or TT genotypes. The frequency of patients carrying both the TBXA2R+795T>C rare allele and HLA DPB1(*)0301 was significantly higher in AIA patients (29.4%) than in ATA patients (7.3%) (P=0.008, odds ratio=5.3). CONCLUSION: These results suggest that the polymorphism of TBXA2R+795T>C may increase bronchoconstrictive response to ASA, which could contribute to the development of the AIA phenotype. PMID- 15898980 TI - Bronchoalveolar lavage eosinophil cationic protein and interleukin-8 levels in acute asthma and acute bronchiolitis. AB - OBJECTIVE: In this study, we measured the levels of eosinophil cationic protein (ECP) and interleukin (IL)-8 in bronchoalveolar lavage (BAL) fluid from patients with acute asthma and acute bronchiolitis, to determine any similarities or dissimilarities in the profiles of these biochemical markers in the two diseases. METHODS: BAL fluids were obtained from children with acute asthma (n=16), infants with acute bronchiolitis caused by respiratory syncytial virus (n=18), and control subjects (n=14). Children with asthma were selected to be free of viral infection. BAL cell counts and differentials were determined, and ECP and IL-8 levels were measured by radioimmunoassay and ELISA, respectively. RESULTS: ECP levels in BAL fluids were significantly higher in the asthma group than in the bronchiolitis (P<0.01) or control (P<0.0001) groups. However, IL-8 levels were significantly higher in the bronchiolitis group than in the asthma (P<0.01) or control (P<0.001) groups. IL-8 levels in the asthma group and ECP levels in the bronchiolitis group were similar to those of the control group. CONCLUSION: This difference in profiles of ECP and IL-8 in acute asthma and acute bronchiolitis, together with a different inflammatory cell pattern, suggests that the nature of the inflammatory process within the lower respiratory tract may be distinctive in these two diseases. PMID- 15898981 TI - Observational study of the natural history of eosinophilic bronchitis. AB - BACKGROUND: Eosinophilic bronchitis is an important cause of chronic cough. Treatment with inhaled corticosteroids is associated with a short-term improvement in cough and reduced sputum eosinophil count but the long-term outcome is uncertain. OBJECTIVE: To determine the long-term outcome in patients diagnosed with and treated for eosinophilic bronchitis. METHODS: We have performed a longitudinal study of symptoms, eosinophilic airway inflammation, spirometry and airway hyper-responsiveness in all patients diagnosed with eosinophilic bronchitis over 7 years. RESULTS: We identified 52 patients with eosinophilic bronchitis and longitudinal data of greater than 1 year (mean 3.1 years) was available in 32 patients, all of whom were treated with inhaled steroids. Three (9%) patients developed symptoms consistent with asthma and a methacholine PC20<8 mg/mL on one or more occasion. Five (16%) patients developed fixed airflow obstruction defined by a persistent post-bronchodilator forced expiratory volume in 1 s (FEV1)/forced vital capacity<70%. One (3%) patient had complete resolution of symptoms and eosinophilic airway inflammation off treatment. The remaining patients had ongoing eosinophilic airway inflammation and/or continuing symptoms. Multiple linear regression identified smoking, female gender and area under the curve of sputum eosinophil count over time as the most important predictors of decline in FEV1. CONCLUSIONS: The most common outcome in eosinophilic bronchitis is continuing disease and complete resolution is rare. Asthma and fixed airflow obstruction developed in relatively few patients. The most important factors associated with a more rapid decline in FEV1 were female gender, smoking and prolonged eosinophilic airway inflammation. PMID- 15898982 TI - Inflammatory cell mapping of the respiratory tract in fatal asthma. AB - BACKGROUND: The site and distribution of inflammation in the airways of asthmatic patients has been largely investigated. Inflammatory cells are distributed in both large and small airways in asthma. It has been demonstrated that distal lung inflammation in asthma may significantly contribute to the pathophysiology of the disease. The upper airways have also been implicated in the overall asthmatic inflammation. Although it is now accepted that lung inflammation is not restricted to the intrapulmonary airways in asthma, little is known about cell distribution in the other lung compartments and their relation to the intrapulmonary airways. OBJECTIVE: We aimed to map the inflammatory process in fatal asthma (FA), from the upper airways to the lung parenchyma. METHODS: Eosinophil, neutrophil, mast cell and lymphocyte content were determined in nasal mucosa, the trachea, intrapulmonary airways and parenchyma (peribronchiolar and distal) of 20 patients with FA and 10 controls. RESULTS: Eosinophil content was higher in all studied areas in FA compared with controls (P<0.02). Mast cell content was higher in the outer area of larger airways, small membranous bronchioles and in peribronchiolar parenchyma of FA compared with controls (P<0.04). CD3+, CD4+and CD20+cells showed increased content in FA intrapulmonary airways compared with controls (P<0.05). There was a positive correlation between CD4+cell content in nasal mucosa and larger airways in asthmatics. Increased neutrophil content was observed only in peribronchiolar parenchyma of FA (P=0.028). CONCLUSION: Eosinophils present a widespread distribution within the respiratory tract in FA, from the nasal mucosa to the distal lung. The outer wall of small membranous bronchioles is the main site of inflammatory changes in FA. There is a localized distribution of alveolar inflammation at the peribronchiolar region for mast cells and neutrophils. Our findings provide further evidence of the importance of the lung periphery in the pathophysiology of FA. PMID- 15898983 TI - Low socioeconomic status as a risk factor for asthma, rhinitis and sensitization at 4 years in a birth cohort. AB - BACKGROUND: The relation between socioeconomic status and allergic diseases in childhood is controversial. Some studies have proposed childhood asthma to be more common in families with low socioeconomic status, while sensitization to airborne allergens seems to be more frequent in individuals with higher socioeconomic status in childhood. OBJECTIVE: To assess the relation between socioeconomic status and asthma, rhinitis and sensitization in an unselected prospective birth cohort. METHODS: Four thousand and eighty-nine families with children born 1994-1996 in predefined areas of Stockholm answered questionnaires on environmental factors, socioeconomic status (parental occupation), and symptoms of allergic disease at birth, 1, 2 and 4 years of age. Blood samples taken at 4 years from 2614 children were analysed for specific IgE to common airborne and food allergens. Odds ratios (OR) and 95% confidence intervals (CI) for various outcomes in relation to socioeconomic status were estimated with a multiple logistic regression model, adjusting for potential confounders such as heredity for allergic diseases, maternal smoking, short duration of breastfeeding and house construction. RESULTS: There was a decreasing risk of asthma and rhinitis with increasing socioeconomic status. The OR for asthma was 0.33 (95% CI 0.17-0.66) and for rhinitis 0.50 (0.32-0.79) comparing the highest and the lowest socioeconomic groups, with a tendency to stronger effects in those with heredity for allergic disease. The risk of sensitization to food allergens also decreased with increasing socioeconomic status; OR 0.65 (0.41-1.02) in the highest socioeconomic group (Ptrend=0.03), which was not clearly seen for airborne allergens. CONCLUSION: Asthma, rhinitis and sensitization is more common in lower than in higher socioeconomic groups after adjustment for traditional risk factors. This may be related to additional uncontrolled differences in life style and environmental exposures between the groups, and calls for further studies. PMID- 15898984 TI - Variability of total serum immunoglobulin E levels from birth to the age of 10 years. A prospective evaluation in a large birth cohort (German Multicenter Allergy Study). AB - BACKGROUND: Many environmental factors influence the concentration of total serum IgE (tIgE); however, tIgE synthesis is believed to be under strong genetic influence. Multiple genetic studies on tIgE regulation have been performed. For these population-based studies tIgE was commonly determined at one time-point, assuming that tIgE phenotypes (adjusted for age and gender) are stable over time. OBJECTIVE: We assessed correlations of tIgE levels from birth to the age of 10 years in the birth cohort MAS (Multicenter Allergy Study; n=1314). MATERIALS AND METHODS: We determined cord-blood IgE levels, total and specific IgE at the age of 1, 2, 3, 5, 6, 7, and 10 years. Spearman correlation coefficients were calculated for tIgE at different time-points. All analyses were performed in the entire cohort, adjusted for gender, as well as in non-atopic children only. RESULTS: tIgE percentiles increased steadily from birth to the age of 10 years with higher values for boys than for girls at each time-point. tIgE values from birth to 3 years of age correlated poorly with tIgE levels at 10 years (r<0.5). However, good correlations (r>0.8) were observed for tIgE concentrations at 6, 7 and 10 years. The same results were observed when the analyses were limited to non-atopic children. CONCLUSION: In childhood, tIgE levels underlie remarkable variation over time even in the absence of atopy. For cross-sectional population based genetic and epidemiologic studies, tIgE values of children <5 years should be interpreted with caution since these values correlate poorly with tIgE levels later in life. PMID- 15898985 TI - Childhood atopy and allergic disease and skin test responses to environmental mycobacteria in rural Crete: a cross-sectional survey. AB - BACKGROUND: Through its powerful immunoregulatory effects, infection with atypical mycobacteria may exert a protective effect on the development of childhood allergic disease. OBJECTIVE: To examine the relationship between childhood atopy or allergic disease and previous infection with four species of atypical mycobacteria. METHODS: Eight hundred and six children aged 8-18 years and living in rural Crete--most of whom had had previous BCG immunization- underwent skin prick testing with 10 aeroallergens; their parents completed a standardized questionnaire relating to allergic disease. No less than 8 weeks later each child underwent intradermal skin tests with 0.1 mL solutions of four selected mycobacterial reagents (Aviumin C, Gordonin, Chelonin and Ranin I). RESULTS: Twenty-three percent of children were atopic on skin prick testing; far fewer had symptoms of asthma (5%) or hayfever in conjunction with a positive prick test to pollens (2%). Eighty percent of children had positive skin responses to one or more mycobacterial species. Among all children--and those with a BCG scar--there was no association between atopy or allergic symptoms and mycobacterial skin responses; among the few children without a BCG scar however those with positive mycobacterial responses were less likely to be atopic or to report allergic symptoms; these differences were not statistically significant. CONCLUSIONS: Our findings, in a population of BCG-immunized children, do not lend support to the suggestion that infection with atypical mycobacteria is protective against childhood allergic disease. PMID- 15898986 TI - The effect of age on the relationship between birth order and immunoglobulin E sensitization. AB - BACKGROUND: An association between birth order and IgE sensitization or allergic diseases has been reported in many studies. OBJECTIVE: To assess the effect of age on the relationship between reduced IgE sensitization and increased birth order and to test the hypothesis that this would decline with increasing age. METHODS: As part of a birth cohort study, IgE sensitization to common allergens was determined by skin prick testing at ages 6 and 12 months, 6 and 11 years. RESULTS: The original cohort numbered 253 individuals of whom 96 (38%) were first born. Compared with individuals with older siblings, first-born individuals had increased IgE sensitization at 6 (odds ratio (OR) 2.4 [95% confidence interval (CI) 1.0, 6.3], P=0.05, n=197) and 12 months of age (OR 6.7 [1.7, 25.0] P=0.002, n=172) and at 6 years of age (OR 2.3 [1.0, 5.6] P=0.05, n=113) but not at 11 years of age (OR 1.2, P>0.4, n=182). When age at onset of IgE sensitization was considered (n=61), 16 had infant onset IgE sensitization (nine were first born), 24 had early childhood onset IgE sensitization (nine first born) and 21 had late childhood onset IgE sensitization (two first born), P=0.0016. Further analysis revealed a similar pattern for children with older brothers (P=0.0097) but not older sisters (P=0.5). CONCLUSIONS: These findings indicated that having an older brother delays the onset of IgE sensitization but may not prevent IgE sensitization per se. The apparent protective effect of older siblings on allergic diseases reported elsewhere might involve delaying the onset of IgE sensitization. PMID- 15898987 TI - Common and distinct signalling cascades in the production of tumour necrosis factor-alpha and interleukin-13 induced by lipopolysaccharide in RBL-2H3 cells. AB - BACKGROUND: Activation of mast cells by lipopolysaccharide (LPS) results in the production of TNF-alpha and IL-13. TNF-alpha and IL-13 are key mediators in the development of neutrophilic and allergic inflammation, respectively. LPS-induced TNF-alpha and IL-13 production in mast cells has been reported to be mediated by Toll-like receptor 4 (TLR4) signalling, but differences in signal transduction mechanisms leading to the production of these cytokines are not clearly defined. OBJECTIVE: We investigated the molecular mechanisms responsible for LPS-induced TNF-alpha and IL-13 production in mast cells. METHODS: TNF-alpha and IL-13 production by LPS was assessed by transfecting RBL-2H3 cells with dominant negative (DN) expression vectors. RESULTS: Transfection of RBL-2H3 cells with plasmids encoding DN mutants of myeloid differentiation protein (MyD88) and TNFR associated factor (TRAF6) inhibited both LPS-induced TNF-alpha and IL-13 production. IkappaBalpha-DN inhibited LPS-induced production of TNF-alpha, but not IL-13. We also found that inhibition of p38 kinase suppressed both TNF-alpha and IL-13 induction by LPS, and inhibition of JNK reduced IL-13 production, but not TNF-alpha. Furthermore, we found that protein kinase R (PKR) was activated by LPS in these cells. Treatment with 2-aminopurine, a PKR inhibitor, attenuated LPS induced nuclear factor-kappaB activation and TNF-alpha production, whereas inhibition of PKR had little effect on IL-13 production. CONCLUSION: These findings indicate that the production of TNF-alpha and IL-13 by LPS required TLR4/MyD88/TRAF6 signalling as a common pathway of mast cell-mediated inflammation. We furthermore found that TNF-alpha and IL-13 production were differentially regulated by signalling cascades through PKR and mitogen-activated protein kinases downstream of TRAF6 in mast cells. PMID- 15898988 TI - Increase in killer-specific secretory protein of 37 kDa in bronchoalveolar lavage fluid of allergen-challenged patients with atopic asthma. AB - BACKGROUND: Atopic asthma is linked to a T-helper type 2 dominated pathogenesis, but there is increasing evidence of Th1/Tc1-mediated processes in the aetiopathology of asthma. Killer-specific secretory protein of 37 kDa (Ksp37) is expressed in cytotoxic lymphocytes, selectively in the effector subsets of CD8+- and CD4+ T lymphocytes and in CD16+/CD56dim natural killer cells and gamma/delta T cells. This effector cell-specific expression of Ksp37 and its coexpression with perforin suggest that Ksp37 might be involved in processes mediated by cytotoxic cells. OBJECTIVE: We hypothesize that Ksp37 could indicate the involvement of cytotoxic lymphocytes in the pathogenesis of atopic asthma, and investigated Ksp37 concentration in bronchoalveolar lavage fluid (BALF) collected 10 min, 18, 42 or 162 h after segmental allergen provocation and in serum of patients with atopic asthma (n=25). METHODS: Ksp37 concentrations in BALF and serum were detected by ELISA. Flow cytometric analysis was used to assess numbers and cell subsets in BALF. RESULTS: Ksp37 increased significantly in BALF 10 min, 18 and 42 h, but not 162 h after allergen challenge compared with saline challenged controls, while Ksp37 serum levels did not change significantly at all time-points. In addition, the increase in Ksp37 concentrations in BALF correlated with the corresponding numbers of lymphocytes. CONCLUSIONS: We conclude that Ksp37 level increased in BALF 10 min, 18 and 42 h after allergen challenge but not in peripheral blood. Our findings suggest that segmental allergen challenge in asthma is associated with an increase in Ksp37 concentrations in BALF and an influx of potentially cytotoxic T lymphocytes into the lungs. PMID- 15898989 TI - The pro-fibrogenic effect of nerve growth factor on conjunctival fibroblasts is mediated by transforming growth factor-beta. AB - BACKGROUND: Nerve growth factor (NGF) and nerve growth factor receptor (NGFR) expressions have been found to be increased in sub-conjunctival scarring. OBJECTIVE: The aim of this study was to investigate the in vitro effects of NGF on some pro-fibrogenic properties of human conjunctival fibroblasts. METHODS: Expression of NGF, trkA(NGFR) and p75NTR on human fibroblasts grown from conjunctival biopsies and incubated for 2 or 6 days with NGF were evaluated by immunofluorescence, RT-PCR, flow cytometry and ELISA. The fibrogenic effect of NGF on conjunctival fibroblasts was investigated by evaluating their migration (wound model), proliferation ([3H]-thymidine incorporation), collagen production (3H]-proline incorporation), expression of alpha-smooth muscle actin (alpha-SMA) (cell surface ELISA) and contraction of 3D collagen gels. RESULTS: NGF induced the expression of p75NTR in the fibroblasts that constitutively expressed only trkA(NGF) and increased the migration of wounded fibroblasts, but not their proliferation and collagen production. NGF induced the conversion of fibroblasts into myofibroblasts expressing alpha-SMA, and enhanced their contraction of a collagen matrix. Interestingly, chronic NGF treatment induced transforming growth factor-beta1 (TGF-beta1) production by fibroblasts, and following specific TGF beta neutralization, all the NGF-induced effects were completely abrogated. CONCLUSION: Our findings indicate that NGF, via TGF-beta induction, is likely to be involved in the healing or fibrotic processes occurring in conjunctiva during some pathological conditions. PMID- 15898990 TI - Rational design of hypoallergens applied to the major cat allergen Fel d 1. AB - BACKGROUND: Allergen-specific immunotherapy is the only treatment for allergic disease providing long-lasting symptom relief. Currently, it is mainly based on the use of crude allergen extracts. The treatment may be improved by the use of genetically engineered allergens, hypoallergens, aiming at a more effective and safer therapy. OBJECTIVE: The aim of this study was to provide a rational design of hypoallergen candidates for immunotherapy by using structural information and knowledge of B and T cell epitopes of an allergen. METHODS: The three-dimensional structure of the major cat allergen Fel d 1 was systematically altered by duplication of selected T cell epitopes and disruption of disulphide bonds. Seven Fel d 1 derivatives were generated and screened for allergenic reactivity in comparison with recombinant Fel d 1 in competition-ELISA. The allergenicity was further evaluated in basophil activation experiments and T cell reactivity was assessed in a lymphoproliferation assay. RESULTS: Three out of seven Fel d 1 derivatives, with two duplicated T cell epitopes and one or two disulphide bonds disrupted, were carefully evaluated. The three derivatives displayed a strong reduction in allergenicity with 400-900 times lower IgE-binding capacity than recombinant Fel d 1. In addition, they induced a lower degree of basophil activation and similar or stronger T cell proliferation than recombinant Fel d 1. CONCLUSION: By a rational approach, we have constructed three Fel d 1 hypoallergens with reduced IgE-binding capacities and retained T cell reactivities. This strategy may be applied to any well-characterized allergen to improve immunotherapy for allergic patients. PMID- 15898991 TI - Identification of human T cell epitopes in Japanese cypress pollen allergen, Cha o 1, elucidates the intrinsic mechanism of cross-allergenicity between Cha o 1 and Cry j 1, the major allergen of Japanese cedar pollen, at the T cell level. AB - BACKGROUND: Pollens from species of Cupressaceae family are one of the most important causes of respiratory allergies worldwide. In Japan, many patients with pollinosis have specific IgE to both pollens of Japanese cypress (Chamaecyparis obtusa) and Japanese cedar (Cryptomeria japonica). The sequences between Cha o 1 and Cry j 1, the major allergens of Japanese cypress and Japanese cedar pollens, respectively, are 80% identical. OBJECTIVE: This study was undertaken to identify T cell epitopes in Cha o 1, and to elucidate the mechanism of cross-allergenicity between Cha o 1 and Cry j 1, at the T cell level. METHODS: T cell epitopes in Cha o 1 were identified by the reactivity of T cell lines, generated from 19 patients, to stimulation with overlapping peptides. The subsets of T cell clones specific to rCha o 1 were determined according to their ability to produce IL-4 and IFN-gamma. Peptide specificities of two T cell clones were determined by stimulation with the peptides from Cha o 1 and Cry j 1. RESULTS: Four dominant and six subdominant T cell epitopes were identified in Cha o 1. While four T cell epitopes, p11-30, p211-230, p251-270 and p331-350, were common to Cha o 1 and Cry j 1, 4 T cell epitopes, p61-80, p71-90, p311-330 and p321-340, were considered to be unique to Cha o 1. The subsets of T cell clones were predominantly of T helper2-type. One T cell clone recognized p16-30, which is common to Cha o 1 and Cry j 1, but another recognized p321-330, which is unique to Cha o 1. CONCLUSION: Presence of both T cells reactive to T cell epitopes common to Cha o 1 and Cry j 1 and T cells specific to T cell epitopes unique to Cha o 1 in patients with pollinosis contributes to prolonged symptoms after the cedar pollen season in March and the following cypress pollen season in April. PMID- 15898992 TI - Persistence of rhinovirus RNA after asthma exacerbation in children. AB - BACKGROUND: Rhinoviruses (RVs) are believed to cause most asthma exacerbations but their role in the severity of acute asthma and subsequent recovery of airway function is not defined. The importance of atopy in virus-host interactions is also not clear. OBJECTIVE: We postulated that RV infection and atopic skin prick responses influence the severity of asthma exacerbations as measured by peak expiratory flow (PEF). METHODS: Patients aged 4-12 years admitted with acute severe asthma to a hospital emergency room (ER) were recruited. PEF measurements were obtained and nasal aspirates (NA) were taken. Atopy was diagnosed by skin prick responses to allergen and the presence of RV RNA and respiratory syncytial virus (RSV) RNA in NAs was detected using validated PCR assays. Patients were restudied after 6 weeks and after 6 months. RESULTS: Fifty children with acute asthma (mean age+/-SD, 7.4+/-2.7) were enrolled; atopy was present in 37 (74%). RV RNA was detected in 41 (82%) and RSV RNA in six (12%) subjects. After 6 weeks 41 patients were restudied and RV RNA was again detected in 18 (44%). RV RNA was detected after 6 months in four of 16 patients restudied (25%; P=0.008 vs. ER) and in two of nine children from a control group with stable asthma (22%; P=0.009 vs. ER). Overall PEF measurements were reduced in asthmatics admitted to ER (% predicted, 63.4+/-16.4%) but did not differ between patients with RV RNA, RSV RNA or neither virus present. In subjects with RV RNA detectable in ER and after 6 weeks, measurements of PEF in ER were significantly lower than in patients in whom RV RNA was present in ER but absent after 6 weeks (P=0.009). Regression analysis linked persistence of RV RNA, but not skin prick responses to allergen, to severity of PEF reductions in ER. CONCLUSION: RV RNA was detectable in >40% of asthmatic children 6 weeks after an acute exacerbation. Asthma exacerbations were more severe in patients with persistence of RV RNA suggesting that the severity of acute asthma may be linked to prolonged and possibly more severe RV infections. PMID- 15898993 TI - Influence of viral infection on the development of nasal hypersensitivity. AB - BACKGROUND: The underlying relationship between viral infections and allergic diseases of the upper respiratory tract has not been well clarified. METHODS: In order to clarify the relationship between viral infection and nasal hypersensitivity, mice were sensitized with ovalbumin (OVA) and then infected intranasally with respiratory syncytial virus (RSV), after which their nasal sensitivity to histamine or antigen was examined. RESULTS: Non-sensitized mice showed transient mild nasal hypersensitivity following nasal administration of histamine after intranasal RSV inoculation. In mice sensitized with OVA, RSV infection significantly exaggerated their nasal hypersensitivity to histamine and OVA. Treatment of these mice with a neurokinin (NK)-1/NK-2 receptor antagonist, but not with anti-IL-5 antibodies, reduced their hypersensitivity. The infiltration of nasal mucosa with eosinophils was temporarily associated with accelerated rate of RSV elimination in these animals. CONCLUSION: RSV infection induced transient nasal hypersensitivity. Several mechanisms, including impairment of nasal epithelial cells are thought to mediate this effect. In allergen-sensitized mice, RSV inoculation strongly enhanced nasal hypersensitivity. PMID- 15898994 TI - Intragastric administration of Mycobacterium vaccae inhibits severe pulmonary allergic inflammation in a mouse model. AB - BACKGROUND: Coexistence with harmless microorganisms such as lactobacilli, saprophytic mycobacteria and some helminths, throughout evolution, may have shaped the host immune system. Exposure to such organisms may have therapeutic benefits by triggering immunoregulatory mechanisms that control inappropriate immune responses to self, gut contents or allergens. OBJECTIVE: We determined whether treatment with Mycobacterium vaccae by gavage influences the host immune response both locally and systemically. We also investigated whether delivery by this route prevents severe symptoms of disease in a murine model of pulmonary allergic inflammation. RESULTS: A single intragastric administration of M. vaccae induced a transient increase in the production of IL-10 and IFN-gamma by mesenteric lymph nodes cells and splenocytes. In addition, in a mouse model of pulmonary allergic inflammation, a single treatment with M. vaccae by gavage not only diminished the total cellular infiltrate and the eosinophilic component induced by subsequent intratracheal allergen challenge, but also biased local and systemic cytokine production towards IL-10. Delivery of M. vaccae by gavage was as effective as subcutaneous treatment. CONCLUSION: This is the first report to suggest that heat-killed mycobacteria can down-regulate symptoms of allergic inflammation by the intragastric route. These data suggest an alternative route of treatment with M. vaccae for patients with allergic conditions. PMID- 15898995 TI - Allergic contact dermatitis from ophthalmic products: can pre-treatment with sodium lauryl sulfate increase patch test sensitivity? AB - In patients suspected of allergic contact dermatitis because of topical ophthalmic medicaments, patch tests performed with patients' own products are often negative. The irritant anionic surfactant sodium lauryl sulfate (SLS) may alter the stratum corneum and increase antigen penetration. Pre-treatment of the skin with SLS 0.5% for 24 h was performed in the sites of patch tests with patients' own products in 15 selected patients. In patients previously negative to their own products tested with conventional patch tests, SLS pre-treatment showed 6 new relevant positive reactions and induced a stronger positive reaction in 1 patient. SLS pre-treatment could be proposed as an alternative promising method, which may increase sensitivity of patch tests with patients' own products. PMID- 15898996 TI - Course of contact allergy in consecutive eczema patients patch tested with TRUE Test panels 1 and 2 at least twice over a 12-year period. AB - Non-reproducibility of positive patch test reactions is common. In this retrospective study, we investigated the reproducibility of previously positive patch test reactions and the appearance of new positive reactions in a consecutive series of 264 eczema patients tested at least twice over a 12-year period using the TRUE Test standardized patch test system. Out of 297 positive reactions, 66% remained positive in a 2nd test, 10% were scored as doubtful and 24% as negative. Among the previously recorded doubtful reactions, 13% were positive in the 2nd test, 13% were again doubtful and 74% negative. In relation to the degree of patch test reactivity, 48% of the + reactions were reproducible, compared to 87% of ++ and +++ reactions. No difference in reproducibility was observed between patients tested with a short or a long-time interval between tests. With the use of the TRUE Test panels 1 and 2, methodological factors are minimized. The variation in patch test positivity may then be attributed to biological variation, which depends on several individual factors such as patch test reading, patient-related factors such as changed immunological sensitivity, regional variation of the skin on the back and the presence of more or less active dermatitis. PMID- 15898997 TI - Transepidermal water loss and incidence of hand dermatitis in a prospectively followed cohort of apprentice nurses. AB - The purpose of the study was to evaluate the incidence of hand dermatitis, the impact of potential risk factors and the efficacy of skin bioengineering in a prospectively followed cohort of apprentice nurses. 104 participants were prospectively followed for 3 years. Before the start of training, after about 1 year and in the third year of occupational exposure, a standardized questionnaire was distributed, and a clinical examination with skin bioengineering of the dorsum of hand and forearm was performed. The 12-month period prevalence of self reported symptoms of hand dermatitis was 36.5%[95%-confidence interval (CI) 27.3 46.6] at intermediate follow-up and 43.3% (95%-CI 33.6-53.3) at the final examination. Apprentices with self-reported symptoms at the final examination showed a significant increase of transepidermal water loss (TEWL) at the dorsal hand from 10.15 g/m2h to 13.55 g/m2h. TEWL at this site did, at the initial examination, not differ significantly between persons who later reported symptoms of hand dermatitis at the final examination and those who did not (10.50 g/m2h versus 10.15 g/m2h, respectively). Our results do not support the notion that an increased basal TEWL is a good indicator for hand dermatitis risk. PMID- 15898998 TI - Incidence and prevalence rates for occupational contact dermatitis in an Australian suburban area. AB - Occupational contact dermatitis (OCD) regularly causes high levels of worker morbidity; however, this is often not reflected in available statistics. This study aimed to collect and verify OCD reports/referrals and generate disease estimates for a defined geographical area in Melbourne, Australia. Two methods of data collection were used. In the first method, 30 general practitioners (GPs), 2 dermatologists and 1 dermatology outpatient clinic within a defined area reported each worker with suspected OCD seen as part of routine practice. With the second method, workers living in the area who were referred to a tertiary referral OCD clinic were included in the study. An occupational dermatologist used a gold standard process that included diagnostic patch testing to verify suspected cases. The incidence rate for confirmed cases was 20.5 per 100,000 workers [95% confidence interval (CI): 13-32.1]. The 1-year-period prevalence rate was 34.5 per 100,000 (95% CI: 24.4-48.7). The positive predictive value (PPV) was highest for the occupational dermatology clinic referrals [63% (95% CI: 49-76%)] compared with reports from the dermatologists/dermatology outpatient clinic [55% (95% CI: 36-74%)] and from GPs [43% (95% CI: 29-59%)]. This study utilizes reports from GPs and dermatologists to provide OCD disease estimates and validation data for an OCD disease register. PMID- 15898999 TI - Deodorants: an experimental provocation study with isoeugenol. AB - Axillary dermatitis is common and overrepresented in people with contact allergy to fragrances. Many people suspect their deodorants to be the incriminating products. In order to investigate the significance of isoeugenol in deodorants for the development of axillary dermatitis when used by people with and without contact allergy to isoeugenol, patch tests with deodorants and ethanol solutions with isoeugenol, as well as repeated open application tests (ROAT) with roll-on deodorants with and without isoeugenol at various concentrations, were performed in 35 dermatitis patients, 10 without and 25 with contact allergy to isoeugenol. A positive ROAT was observed only in patients hypersensitive to isoeugenol (P<0.001) and only in the axilla to which the deodorants containing isoeugenol had been applied (P<0.001). Deodorants containing isoeugenol in the concentration range of 0.0063-0.2% used 2 times daily on healthy skin can thus elicit axillary dermatitis within a few weeks in people with contact allergy to isoeugenol. PMID- 15899000 TI - Potency and risk assessment of a skin-sensitizing disperse dye using the local lymph node assay. AB - Disperse Blue 106 is an acknowledged skin-sensitization hazard. However, information about the relative sensitization potency of this chemical is lacking, and to provide this information was the purpose of the investigations described here. The approach taken was to measure dose-response relationships for C.I. Disperse Blue in the local lymph node assay, a method for the assessment of skin sensitization potential in which activity is measured as a function of lymphocyte proliferative responses induced in draining lymph nodes. From these data, it was possible to derive EC3 values (such being the estimated concentration of chemical required to elicit a 3-fold increase in proliferation) that have been shown previously to reflect the relative sensitizing potency of contact allergens. These analyses revealed that Disperse Blue 106 had a relatively low EC3 value (0.01%), comparable to that measured concurrently for 2,4-dinitrochlorobenzene, a potent contact allergen. Collectively, these data reveal that Disperse Blue 106 represents a significant skin-sensitization hazard, and, in combination with information on dye migration and percutaneous penetration from various types of fabric and use conditions provide a basis for the development of effective and accurate risk assessments. PMID- 15899001 TI - Peristomal allergic contact dermatitis--case report and review of the literature. AB - Allergic contact dermatitis (ACD) is a rare cause of peristomal skin problems. Only 23 cases have previously been reported in the literature. We report the case of a colostomy patient with a severe disabling blistering peristomal dermatitis. Patch testing to a British Contact Dermatitis Society standard series, medicaments and a plastics and glues series was negative. Patch testing to the patient's own products gave a positive reaction (+) at D2 and D4 to Dansac soft paste and Stomahesive paste. Further patch testing to the components of Dansac soft paste showed a positive (+) reaction at D2 and D4 to ester of polymethyl vinyl/maleic acid copolymer (Gantrez-ES) only. This is the first reported case of ACD due to Dansac soft paste. ACD to Gantrez has previously been reported but in different products. We also review the other previously reported cases of ACD causing peristomal dermatitis and stress the importance of patch testing in these cases, in particular to the patient's own products, as avoidance of identified allergens can have a large impact on the quality of life. PMID- 15899002 TI - Occupational contact allergy to glyoxal. AB - Glyoxal is a dialdehyde that is used as a disinfectant in health care and dentistry work. Allergic contact dermatitis from glyoxal has been described in these occupations. We analysed our patient data from 1998 to 2004 for allergic reactions to glyoxal. 20 patients had allergic reactions to glyoxal on patch testing. 5 of these patients worked in dentistry and 4 of them had present exposure to glyoxal. 9 patients were machinists without obvious exposure to glyoxal. A grinder with work-related facial dermatitis is described in detail. The chemical analysis of air samples from his workplace revealed 9.4-21 microg/m3 glyoxal. Glyoxal was also present in the used metal-working fluid, and apparently it had been formed during grinding. The remaining 6 patients worked in miscellaneous occupations and had no present exposure to glyoxal. Glyoxal is irritant on patch testing. Especially, solitary reactions to glyoxal 10% in aq. may be false-positive irritant reactions. 9 (45%) of our patients reacted to formaldehyde or glutaraldehyde. Glyoxal is an important allergen in dentistry and medical care, and we recommend it to be added to the antimicrobial patch test series. It also seems to be a 'hidden' allergen in the metal industry. PMID- 15899003 TI - Severe delayed cutaneous reaction due to Mediterranean jellyfish (Rhopilema nomadica) envenomation. AB - During summer, Mediterranean Sea waters are invaded by a species of jellyfish designated as Rhopilema nomadica. Their tentacles contain numerous nematocysts loaded with a toxin that causes envenomation, usually expressed as immediate appearance of redness, burning sensation and papulovesicular eruption in the affected skin. We report a lady with a severe delayed reaction due to jellyfish envenomation that developed 2 days after contact with jellyfish tentacles. PMID- 15899004 TI - Immediate cutaneous hypersensitivity response to phytomenadione induced by vitamin K in skin diagnostic procedure. PMID- 15899005 TI - Meat protein allergic contact dermatitis: A case report. PMID- 15899006 TI - Occupational contact dermatitis from simvastatin. PMID- 15899007 TI - Lichen dermatitis in a rock climber. PMID- 15899008 TI - Sensitization to para-phenylenediamine in a streetside temporary tattoo artisan. PMID- 15899009 TI - The baboon syndrome--report of two first cases in Poland. PMID- 15899010 TI - Two cases of occupational allergic contact dermatitis to p-aminophenol in pharmaceutical workers manufacturing paracetamol. PMID- 15899011 TI - Hand dermatitis among a complete cross-section of Chinese physicians. PMID- 15899012 TI - Detection of sensitivity to cardol in north-eastern and south-eastern Brazil. PMID- 15899013 TI - Unusual allergen in nurse uniform trousers. PMID- 15899014 TI - Thresholds of contact sensitization from disperse dyes in textiles. PMID- 15899016 TI - Acupuncture in xerostomia--a systematic review. AB - The aim of this systematic review was to assess the efficacy of acupuncture in the management of xerostomia. Assessing quality of studies aim to efficiently integrate valid information and provide a basis for sound decision making based on the best available evidence. Articles of controlled clinical trials evaluating the efficacy of acupuncture in the management of xerostomia were obtained by searching through the databases MEDLINE and Cochrane Central Register of Controlled Trials to September 2003. Three articles met the criteria for inclusion and a criteria list was used to assess the quality of these studies. The studies were considered to be of high quality or low quality in accordance with the criteria list utilized. The results of the trials were considered positive, negative or indifferent based on statistically significant between group differences. The criteria list utilized indicate that one of the three studies was of high quality and it presents indifferent results. One of the two studies of low quality presents positive results and one presents indifferent results. An analysis of the results degree of evidence resulted in no evidence for the efficacy of acupuncture in the management of xerostomia. This systematic review shows that there is no evidence for the efficacy of acupuncture in the management of xerostomia. There is a need for future high quality randomized controlled trials. PMID- 15899017 TI - Factors related to the resting tongue position among partially and completely edentulous subjects. AB - The prevalence of abnormal (retracted) position of the tongue at rest was examined in subjects with varying number of natural lower teeth. The aim of the study was to explore the relationship between tongue retraction and state of dentition and to identify potentially related parameters. The resting tongue positions were recorded in subjects partially edentulous in the lower jaw (n = 164) and compared with those met in dentate (n = 57) and completely edentulous participants (n = 84). Potentially related parameters, such as age, sex, duration of edentulism, palate vault and signs of temporomandibular disorders were also recorded. Tongue retraction was observed in a small percentage (12.3%) of the dentate, almost half (45.7%) of the partially edentulous and in the majority (67.8%) of the completely edentulous participants. The number of natural lower teeth correlated with the resting tongue position in the partially edentulous group. The tongue position at rest was also found weakly related to signs of temporomandibular disorders in the dentate group and to the sex and duration of edentulism in the partially edentulous group. It was concluded that the abnormal positioning of the resting tongue is increasingly observed with decreasing number of natural teeth, possibly in accordance with the morphological and functional alterations that result from teeth loss. The presence of a retracted tongue affects the complete denture construction; however, its effect on denture function remains questionable. PMID- 15899018 TI - Masticatory function in patients with an extremely resorbed mandible restored with mandibular implant-retained overdentures: comparison of three types of treatment protocols. AB - The objective of the present study was to analyse the effects of implant supported overdentures on masticatory function in patients with an extremely resorbed mandible, and to compare the masticatory function in these patients using three differing types of implant treatment protocols. The mandibular overdentures were retained by a transmandibular implant, by four endosseous implants following augmentation of the mandible, and by four short endosseous implants, respectively. Sixty patients (50 women, 10 men, mean age 59.4 years) were randomly allocated to one of the three treatment groups. Masticatory function was assessed before and after treatment using a questionnaire, a masticatory performance test, and a structured interview. The patient-based masticatory function improved significantly. Concerning these parameters there were no significant differences between the three groups before and after treatment. A significant difference existed between the three groups for the laboratory-assessed masticatory function before treatment, but after treatment this difference was no longer significant. From this study it can be concluded that patients with an extremely resorbed mandible and functional complaints of their lower denture report significant improvement in masticatory function after implant-overdenture treatment. Differences in masticatory function between the three studied modalities were not significant after treatment. PMID- 15899019 TI - Surface EMG of jaw elevator muscles: effect of electrode location and inter electrode distance. AB - This study addresses methodological issues on surface electromyographic (EMG) signal recording from jaw elevator muscles. The aims were (i) to investigate the sensitivity to electrode displacements of amplitude and spectral surface EMG variables, (ii) to analyse if this sensitivity is affected by the inter-electrode distance of the bipolar recording, and (iii) to investigate the effect of inter electrode distance on the estimated amplitude and spectral EMG variables. The superficial masseter and anterior temporalis muscles of 13 subjects were investigated by means of a linear electrode array. The percentage difference in EMG variable estimates from signals detected at different locations over the muscle was larger than 100% of the estimated value. Increasing the inter electrode distance resulted in a significant reduction of the estimation variability because of electrode displacement. A criterion for electrode placement selection is suggested, with which the sensitivity of EMG variables to small electrode displacements was of the order of 2% for spectral and 6% for amplitude variables. Finally, spectral and, in particular, amplitude EMG variables were very sensitive to inter-electrode distance, which thus should be fixed when subjects or muscles are compared in the same or different experimental conditions. PMID- 15899020 TI - The effects of phosphoric acid, sodium hypochlorite, ferric oxalate and Scotchbond Multipurpose on the rate of pressure change across dentine: a laboratory study. AB - The aim of this study was to investigate the consequences of dentine treatment on dentine permeability of crown preparations treated with phosphoric acid, sodium hypochlorite (NaOCl), and then ferric oxalate (Sensodyne sealant) or bis-phenol-A diglycidyldimethacrylate (BIS-GMA) and hydroxyethylmethacrylate (HEMA) resin combination (Scotchbond Multipurpose). Thirty human premolars received full crown preparations (stage A), were then acid etched (stage B) and subsequently the pulp chamber flushed with NaOCl (stage C). They were then randomly assigned to two groups: group 1 was treated with Scotchbond Multipurpose and group 2 with 6% ferric oxalate (Sensodyne sealant) (stage D). Both groups were acid etched a second time (stage E) as the final step in the sequence. The rise in pressure in the pulp chamber because of the transmission of saline from the pressure chamber through cut dentine was recorded by a pressure transducer after each of the stages described (A to E). The mean rate of pressure change across dentine (Pa s( 1)) for each measurement point (A to E) (n = 30) were A = 2.3; B = 9.8; C = 16; D = 2.1; E = 3.1. Acid etching and NaOCl were both effective in producing significant increases in the rate of pressure change across dentine (P < 0.05). Sensodyne sealant and Scotchbond Multipurpose are both effective dentine sealants and there is no significant difference (P > 0.05) in their abilities in sealing dentine. Acid etching-sealed dentine produced a significant increase in the rate of pressure change across dentine (P < 0.05). The sealing of dentine appears to be a sensible consideration following crown preparations on vital teeth. PMID- 15899021 TI - An experimental analysis of stresses in simulated flared root canals subjected to various post-core applications. AB - Cervical root fracture is a major clinical problem in flared root canals treated with different post and core applications. This in vitro study evaluated the stress transfer of different post and core systems to the cervical part of the artificially created flared root canals, by using strain gauges. The post-core systems investigated were: (a) cast post-core system without resin reinforcement, (b) cast post-core system with resin reinforcement, (c) pre-fabricated post and resin core with resin reinforcement. The post-core systems which were cemented on simulated roots were subjected to a load applied at an angle of 45 degrees to the long axis of the simulated roots. The strain gauges which were cemented to the cervical part of simulated roots were connected to the data acquisition module to measure and record the changes in strain data. Specimens restored with resin reinforcement either with cast post-core or pre-fabricated post and resin core transferred the stress to the cervical part of the artificial roots at a rate lower than conventional cast post-core system (P = 0.001; anova and Tukey's post hoc test). It was concluded that the resin reinforcement of root canals before post-core applications reduces the stresses at the cervical part of the root surfaces. PMID- 15899022 TI - Effect of luting agents and reconstruction techniques on the fracture resistance of pre-fabricated post systems. AB - Fracture resistance and fracture modes of endodontically treated maxillary central incisors restored with different post-and-core systems covered with all ceramic copings were evaluated. Ten samples were prepared for each group. Groups 1, 2 and 3 consisted of tooth-coloured post-and-core, zirconia post (Cosmopost) with a composite core (Tetric Ceram), zirconia post (Cosmopost) with a custom made ceramic core (Cosmo Ingot), glass fibre-reinforced post (FRC Postec) with a composite core (Tetric Ceram), respectively. Group 4 consisted of a titanium post (ERpost) with a composite core (Tetric Ceram). The control group (group 5) consisted of root-filled incisors without posts. Tooth-coloured posts were cemented in the roots using Variolink-2, while titanium posts were cemented in the roots using Harvard cement. The all-ceramic copings were cemented using Variolink-2. Static load was applied to 2 mm below the incisal edge on the palatinal surface of each sample until they were fractured. Fracture data were obtained and statistically analysed with One-way anova and a Tukey's test. The results of the means and standard deviations of the fracture resistance during static loading were: 497.5 +/- 61.94 (1), 474.61 +/- 96.84 (2), 494.61 +/- 104.67 (3), 581.34 +/- 105.36 (4), 420.42 +/- 127.48 (5). There were statistically significant differences between groups 4 and 5. Glass fibre-reinforced posts and composite cores (group 3) showed the most catastrophic failure. Consequently, zirconia ceramic posts can be used in clinical practice. PMID- 15899023 TI - Marginal and internal fit of all-ceramic CAD/CAM crown-copings on chamfer preparations. AB - Evaluation of the marginal and internal fit of all-ceramic molar crown-copings hypothesizing that Computer Aided Design/Computer Aided Manufacturing (CAD/CAM) fabrication shows the same accuracy of fit as conventional techniques. A set of six individual crown preparations was duplicated 12 times yielding 72 plaster dies. Slip-cast (In-Ceram Zirconia), heat-pressing (Empress II) and CAD/CAM crown copings (Cerec inLab, DCS, Decim and Procera) were seated on 12 dies each. Marginal and internal gap width was measured in the SEM at 120x magnification. Marginal gap of slip-cast (25 +/- 18 microm) was significantly (P < 0.05) smaller than that of Empress II (44 +/- 23 microm) copings. Procera (17 +/- 16 microm) and Decim (23 +/- 17 microm) did not differ (P > 0.05) from slip-cast (25 +/- 18 microm) but were smaller (P < 0.001/P < 0.01) than Empress II (44 +/- 23 microm) and Cerec inLab (43 +/- 23 microm) (P < 0.001/P < 0.05). DCS (33 +/- 20 microm) did not differ (P > 0.05) from any of the others. The internal mid-orobuccal gap width of Procera (136 +/- 68 microm) was larger (P < 0.001) than that of Decim (81 +/- 30 microm) and slip-cast (94 +/- 84 microm) (P < 0.05) while Empress II (105 +/- 53 microm), DCS (110 +/- 79 microm) and Cerec inLab (114 +/- 58 microm) did not differ significantly (P > 0.05) from Decim, Procera and slip-cast. Internal mesiodistal gap width was similar. The fit of conventional and CAD/CAM all-ceramic molar crown-copings covered the same range of gap width confirming the assumed hypothesis. PMID- 15899024 TI - Evaluation of internal adaptation in ceramic and composite resin inlays by silicon replica technique. AB - This study was aimed at investigating the internal adaptation of a ceramic (Ceramco II) and two composite resin inlay materials (SureFil and 3M Filtek Z 250) using silicon replica technique as an indicator. Forty-five standard mesial occlusal-distal (MOD) cavities were prepared into brass moulds by using computer numerically controlled system. Inlays were prepared according to manufacturers' instructions with indirect methods. Replicas of the prepared cavities and inlays were produced with a polyvinyl siloxane material (Elite H-D). The spaces between inlays and cavities were filled by different coloured light-body polyvinyl siloxane material. Two parallel slices (mesio-distally) were obtained from the replicas with a sharp blade. Different coloured polyvinyl siloxane material thickness between cavity and inlay was measured at seven points (mesial, occlusal and distal). The data were evaluated with anova and Tukey's honestly significantly different (HSD) statistical tests. In the SureFil and Ceramco II groups, the sizes of the contraction gaps at mesial and distal gingival floors were greater than that of the occlusal marginal walls. In comparison of gap formation at occlusal regions, while the 3M composite group showed highest gap values (204.33 +/- 75.45 microm), the Ceramco II group revealed the lowest (141.17 +/- 23.66 microm) (P < 0.05). At the gingival floors, gap formation of Ceramco II group was the highest (227.08 +/- 51.95 microm). Neither the 3M Filtek Z250 nor SureFil group showed any statistical difference between gap values of their self-occlusal and gingival floors (P > 0.05). In conclusion, our results showed that ceramic inlays did not confer any big advantage for internal adaptation over the composite inlays. PMID- 15899025 TI - In vitro evaluation of the bonding of auto-polymerizing soft denture liner to cobalt-chromium alloy. AB - This study evaluated the effects of surface pre-treatments on the bonding of auto polymerizing silicone soft denture liner to a Co-Cr alloy denture base after cyclic thermal stressing. The bonding surfaces of Co-Cr alloy cylinders (8 mm diameter and 4 mm high) were polished with a 600-grit silicon carbide paper. The bonding surfaces received one of three pre-treatments, which included either the application of a metal primer, a metal primer after air abrasion, or a resin primer after adhesive resin coating. Tensile specimens were fabricated by polymerizing a 2-mm thickness of soft denture liner between a pair of pre-treated denture base cylinders. Failure loads were measured by tensile testing after subjecting the specimen to 0, 5, 10, 20, and 30 thousand thermal cycles. Seven specimens were fabricated for 15 groups, including three pre-treatments and five thermal cycle groups. Failure loads of resin-bonded specimens, which were formerly reported, were used as references. All data were statistically analysed by two-way anova and Bonferroni test at the 95% confidence level. Adhesive resin coating of the metal surface was effective in enhancing the failure loads up to 5000 thermal cycles. However, failure loads of all metal-bonded groups were significantly lower than those of resin-bonded groups at all thermal cycling intervals. The results of this in vitro study implied that polymerizing adhesive resin on the Co-Cr alloy might be a promising method, when the clinicians need to acquire improved bonding of an auto-polymerizing soft denture liner to Co-Cr alloy denture bases. PMID- 15899026 TI - Pulmonary aspiration of a two-unit bridge during a deep sleep. AB - Aspiration of teeth and dental restorations is a recognized, yet an infrequent happening in the literature. Main reasons of aspiration are maxillofacial trauma, dental treatment procedures or ethanol intoxication and dementia. The present case of a 2-unit bridge aspiration is however, not related with any trauma, dental procedure or systemic disease. A 37-year-old male patient had aspirated his bridge while sleeping and the bridge remained unidentified for 1 year despite the radiographic controls. He was then referred to the Chest Diseases Department of School of Medicine, Ege University and the radio-opaque object in the right intermediate bronchus was diagnosed to be an aspirated dental prosthesis. Subsequent to the failure of the rigid bronchoscopy, the patient was referred to the Thoracic Surgery Department and had to be operated for retrieval of the foreign body. PMID- 15899027 TI - Occlusion--time to reflect? A personal view. AB - The topic of occlusion presents a conundrum to many practitioners. To some the topic is very complex and yet to others, although important, it illustrates how adaptive the dentition is to change. This commentary presents some arguments as to how the importance of occlusion and its understanding have evolved since the Journal of Oral Rehabilitation was first published. PMID- 15899028 TI - Levels of gastrin-releasing peptide and substance P in synovial fluid and serum correlate with levels of cytokines in rheumatoid arthritis. AB - It is well known that cytokines are highly involved in the disease process of rheumatoid arthritis (RA). Recently, targeting of neuropeptides has been suggested to have potential therapeutic effects in RA. The aim of this study was to investigate possible interrelations between five neuropeptides (bombesin/gastrin-releasing peptide (BN/GRP), substance P (SP), vasoactive intestinal peptide, calcitonin-gene-related peptide, and neuropeptide Y) and the three cytokines tumour necrosis factor (TNF)-alpha, IL-6, and monocyte chemoattractant protein-1 in synovial fluid of patients with RA. We also investigated possible interrelations between these neuropeptides and soluble TNF receptor 1 in serum from RA patients. Synovial fluid and sera were collected and assayed with ELISA or RIA. The most interesting findings were correlations between BN/GRP and SP and the cytokines. Thus, in synovial fluid, the concentrations of BN/GRP and SP grouped together with IL-6, and SP also grouped together with TNF-alpha and monocyte chemoattractant protein-1. BN/GRP and SP concentrations in synovial fluid also grouped together with the erythrocyte sedimentation rate. In the sera, BN/GRP concentrations and soluble TNF receptor 1 concentrations were correlated. These results are of interest because blocking of SP effects has long been discussed in relation to RA treatment and because BN/GRP is known to have trophic and growth-promoting effects and to play a role in inflammation and wound healing. Furthermore, the observations strengthen a suggestion that combination treatment with agents interfering with neuropeptides and cytokines would be efficacious in the treatment of RA. In conclusion, BN/GRP and SP are involved together with cytokines in the neuroimmunomodulation that occurs in the arthritic joint. PMID- 15899029 TI - The utility of pathway selective estrogen receptor ligands that inhibit nuclear factor-kappa B transcriptional activity in models of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease that produces synovial proliferation and joint erosions. The pathologic lesions of RA are driven through the production of inflammatory mediators in the synovium mediated, in part, by the transcription factor NF-kappaB. We have identified a non steroidal estrogen receptor ligand, WAY-169916, that selectively inhibits NF kappaB transcriptional activity but is devoid of conventional estrogenic activity. The activity of WAY-169916 was monitored in two models of arthritis, the HLA-B27 transgenic rat and the Lewis rat adjuvant-induced model, after daily oral administration. In both models, a near complete reversal in hindpaw scores was observed as well as marked improvements in the histological scores. In the Lewis rat adjuvant model, WAY-169916 markedly suppresses the adjuvant induction of three serum acute phase proteins: haptoglobin, alpha1-acid glycoprotein (alpha1-AGP), and C-reactive protein (CRP). Gene expression experiments also demonstrate a global suppression of adjuvant-induced gene expression in the spleen, liver, and popliteal lymph nodes. Finally, WAY-169916 was effective in suppressing tumor necrosis factor-alpha-mediated inflammatory gene expression in fibroblast-like synoviocytes isolated from patients with RA. Together, these data suggest the utility of WAY-169916, and other compounds in its class, in treating RA through global suppression of inflammation via selective blockade of NF-kappaB transcriptional activity. PMID- 15899030 TI - Clinical response to discontinuation of anti-TNF therapy in patients with ankylosing spondylitis after 3 years of continuous treatment with infliximab. AB - We analyzed the clinical response and the time to relapse after discontinuation of continuous long-term infliximab therapy in patients with ankylosing spondylitis (AS). After 3 years of infliximab therapy, all AS patients (n = 42) discontinued treatment (time point (TP)1) and were visited regularly for 1 year in order to assess the time to relapse (TP2). Relapse was defined as an increase to a value >or= 4 on the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and a physician's global assessment >or= 4 according to the recommendations of the Assessments in Ankylosing Spondylitis (ASAS) working group. After 52 weeks, 41 of the 42 patients (97.6%) had to be reinfused because of relapse. The mean change in the BASDAI between TP1 and TP2 was 3.6 +/- 1.7 and that in the physician's global assessment was 4.4 +/- 1.8 (both P < 0.001). The mean time to relapse was 17.5 weeks (+/- 7.9 weeks, range 7 to 45). Ten patients (24%) showed a relapse within 12 weeks and 38 patients (90.5%), within 36 weeks. After 52 weeks, only one patient had remained in ongoing remission without further treatment with anti-tumor-necrosis factor. Patients who were in partial remission according to the ASAS criteria and those with normal C-reactive protein levels at the time point of withdrawal had longer times to relapse after discontinuation of the treatment. Retreatment with infliximab was safe and resulted in clinical improvement in all patients to a state similar to that before the treatment was stopped. Discontinuation of long-term therapy with infliximab eventually led to relapse of disease activity in all patients but one. PMID- 15899031 TI - Expression of cytokine mRNA and protein in joints and lymphoid organs during the course of rat antigen-induced arthritis. AB - Cytokine expression was assessed during antigen-induced arthritis (AIA) in synovial membrane (SM), inguinal lymph node (LN), and spleen using competitive RT PCR and sandwich ELISA. In the SM, early elevations of IL-1beta and IL-6 mRNA (by 6 hours; 450- and 200-fold, respectively) correlated with the joint swelling; a 6 fold increase in tumor necrosis factor alpha (TNFalpha) was not significant. Not only IL-2 and IFN-gamma (which increased 10,000-fold and 200-fold, respectively), but also IL-5 and IL-10, increased acutely (6 hours - day 1; 3-fold and 35-fold, respectively) in the SM. In general, the protein levels in the SM for IL-1beta, IL-6, TNFalpha, IFN-gamma, IL-4, and IL-10 (increase from 4-fold to 15-fold) matched the course of mRNA expression. In the inguinal LN, there were early mRNA elevations of IL-6 (a 2.5-fold increase by 6 hours, which correlated positively with the joint swelling) and IL-2 (4-fold by 6 hours), as well as later rises of IL-4 and IL-5 (2.5- and 4-fold, respectively, by day 3). No significant elevations of the corresponding proteins in this tissue were observed, except for IL-1beta (by day 6) and IL-10 (by day 1). In the spleen, there were significant mRNA elevations at 6 hours of IL-1beta (1.5-fold), IL-6 (4-fold; positively correlated with the joint swelling), IFN-gamma (3-fold), and IL-2 (7- to 10 fold). IL-5 and IL-10 (2- and 3-fold, respectively) peaked from 6 hours to day 3 in the spleen. Increases of the corresponding proteins were significant in comparison with day 0 only in the case of IL-2 (day 6). By day 6 (transition to the chronic phase), the mRNA for cytokines declined to or below prearthritis levels in all the tissues studied except for IL-1beta in the SM and IL-6 in the spleen. AIA is thus characterized by four phenomena: early synovial activation of macrophages, T helper (Th)1-like, and Th2-like cells; late, well-segregated Th2 like responses in the inguinal LN; late, overlapping Th1-like/Th2-like peaks in the spleen; and chronic elevation of synovial IL-1beta mRNA and spleen IL-6 mRNA. PMID- 15899032 TI - Citrullinated proteins have increased immunogenicity and arthritogenicity and their presence in arthritic joints correlates with disease severity. AB - Autoantibodies directed against citrulline-containing proteins have an impressive specificity of nearly 100% in patients with rheumatoid arthritis and have been suggested to be involved in the disease pathogenesis. The targeted epitopes are generated by a post-translational modification catalysed by the calcium-dependent enzyme peptidyl arginine deiminase (PAD), which converts positively charged arginine to polar but uncharged citrulline. The aim of this study was to explore the effects of citrullination on the immunogenicity of autoantigens as well as on potential arthritogenicity. Thus, immune responses to citrullinated rat serum albumin (Cit-RSA) and to unmodified rat serum albumin (RSA) were examined as well as arthritis development induced by immunisation with citrullinated rat collagen type II (Cit-CII) or unmodified CII. In addition, to correlate the presence of citrullinated proteins and the enzyme PAD4 with different stages of arthritis, synovial tissues obtained at different time points from rats with collagen induced arthritis were examined immunohistochemically. Our results demonstrate that citrullination of the endogenous antigen RSA broke immunological tolerance, as was evident by the generation of antibodies directed against the modified protein and cross-reacting with the native protein. Furthermore we could demonstrate that Cit-CII induced arthritis with higher incidence and earlier onset than did the native counterpart. Finally, this study reveals that clinical signs of arthritis precede the presence of citrullinated proteins and the enzyme PAD4. As disease progressed into a more severe and chronic state, products of citrullination appeared specifically in the joints. Citrullinated proteins were detected mainly in extracellular deposits but could also be found in infiltrating cells and on the cartilage surface. PAD4 was detected in the cytoplasm of infiltrating mononuclear cells, from day 21 after immunisation and onwards. In conclusion, our data reveal the potency of citrullination to break tolerance against the self antigen RSA and to increase the arthritogenic properties of the cartilage antigen CII. We also show that citrullinated proteins and the enzyme PAD4 are not detectable in healthy joints, and that the appearance and amounts in arthritic joints of experimental animals are correlated with the severity of inflammation. PMID- 15899033 TI - The P2X7 receptor is a candidate product of murine and human lupus susceptibility loci: a hypothesis and comparison of murine allelic products. AB - Systemic lupus erythematosus and its murine equivalent, modelled in the New Zealand Black and New Zealand White (NZB x NZW)F1 hybrid strain, are polygenic inflammatory diseases, probably reflecting an autoimmune response to debris from cells undergoing programmed cell death. Several human and murine loci contributing to disease have been defined. The present study asks whether the proinflammatory purinergic receptor P2X7, an initiator of a form of programmed cell death known as aponecrosis, is a candidate product of murine and human lupus susceptibility loci. One such locus in (NZB x NZW)F1 mice is lbw3, which is situated at the distal end of NZW chromosome 5. We first assess whether NZB mice and NZW mice carry distinct alleles of the P2RX7 gene as expressed by common laboratory strains, which differ in sensitivity to ATP stimulation. We then compare the responses of NZB lymphocytes, NZW lymphocytes and (NZB x NZW)F1 lymphocytes to P2X7 stimulation. NZB and NZW parental strains express the distinct P2X7-L and P2X7-P alleles of P2RX7, respectively, while lymphocytes from these and (NZB x NZW)F1 mice differ markedly in their responses to P2X7 receptor stimulation. NZB mice and NZW mice express functionally distinct alleles of the proinflammatory receptor, P2X7. We show that current mapping suggests that murine and human P2RX7 receptor genes lie within lupus susceptibility loci lbw3 and SLEB4, and we argue that these encode a product with the functional characteristics consistent with a role in lupus. Furthermore, we argue that aponecrosis as induced by P2X7 is a cell death mechanism with characteristics that potentially have particular relevance to disease pathogenesis. PMID- 15899034 TI - Small GTP-binding protein Rho-mediated signaling promotes proliferation of rheumatoid synovial fibroblasts. AB - Rho is a major small GTP-binding protein that is involved in the regulation of various cell functions, including proliferation and cell migration, through activation of multiple signaling molecules in various types of cells. We studied its roles in synovial fibroblasts (SFs) in patients with rheumatoid arthritis (RA) and clarified its relevance to RA synovitis, with the following results. 1)We found that the thrombin receptor was overexpressed on RA synovial fibroblasts (RA SFs) and that thrombin induced a marked proliferation and progression of the cell cycle to the S phase in these cells. 2)We also found that thrombin efficiently activated Rho. 3)Rho activation and proliferation and the progression of the cell cycle to the S phase were completely blocked by p115RGS (an N-terminal regulator of the G-protein signaling domain of p115RhoGEF) and by the C-terminal fragments of Galpha13 (an inhibitor of the interaction of receptors with G13). 4)Thrombin induced the secretion of IL-6 by RA SFs, but this action was blocked by p115RGS or Galpha13. Our findings show that the actions of thrombin on the proliferation of RA SFs, cell-cycle progression to the S phase, and IL-6 secretion were mainly mediated by the G13 and RhoGEF pathways. These results suggest that p115RGS and Galpha13 could be potent inhibitors of such functions. A rational design of future therapeutic strategies for RA synovitis could perhaps include the exploitation of the Rho pathway to directly reduce the growth of synovial cells. PMID- 15899035 TI - A web tool for finding gene candidates associated with experimentally induced arthritis in the rat. AB - Rat models are frequently used for finding genes contributing to the arthritis phenotype. In most studies, however, limitations in the number of animals result in a low resolution. As a result, the linkage between the autoimmune experimental arthritis phenotype and the genomic region, that is, the quantitative trait locus, can cover several hundred genes. The purpose of this work was to facilitate the search for candidate genes in such regions by introducing a web tool called Candidate Gene Capture (CGC) that takes advantage of free text data on gene function. The CGC tool was developed by combining genomic regions in the rat, associated with the autoimmune experimental arthritis phenotype, with rat/human gene homology data, and with descriptions of phenotypic gene effects and selected keywords. Each keyword was assigned a value, which was used for ranking genes based on their description of phenotypic gene effects. The application was implemented as a web-based tool and made public at http://ratmap.org/cgc. The CGC application ranks gene candidates for 37 rat genomic regions associated with autoimmune experimental arthritis phenotypes. To evaluate the CGC tool, the gene ranking in four regions was compared with an independent manual evaluation. In these sample tests, there was a full agreement between the manual ranking and the CGC ranking for the four highest-ranked genes in each test, except for one single gene. This indicates that the CGC tool creates a ranking very similar to that made by human inspection. The exceptional gene, which was ranked as a gene candidate by the CGC tool but not in the manual evaluation, was found to be closely associated with rheumatoid arthritis in additional literature studies. Genes ranked by the CGC tools as less likely gene candidates, as well as genes ranked low, were generally rated in a similar manner to those done manually. Thus, to find genes contributing to experimentally induced arthritis, we consider the CGC application to be a helpful tool in facilitating the evaluation of large amounts of textual information. PMID- 15899036 TI - Peripheral blood but not synovial fluid natural killer T cells are biased towards a Th1-like phenotype in rheumatoid arthritis. AB - Natural killer T (NKT) cells have been implicated in the regulatory immune mechanisms that control autoimmunity. However, their precise role in the pathogenesis of rheumatoid arthritis (RA) remains unclear. The frequency, cytokine profile and heterogeneity of NKT cells were studied in peripheral blood mononuclear cells (PBMCs) from 23 RA patients and 22 healthy control individuals, including paired PBMC-synovial fluid samples from seven and paired PBMC-synovial tissue samples from four RA patients. Flow cytometry revealed a decreased frequency of NKT cells in PBMCs from RA patients. NKT cells were present in paired synovial fluid and synovial tissue samples. Based on the reactivity of PBMC-derived NKT cells toward alpha-galactosylceramide, RA patients could be divided into responders (53.8%) and nonresponders (46.2%). However, NKT cells isolated from synovial fluid from both responders and nonresponders expanded upon stimulation with alpha-galactosylceramide. Analysis of the cytokine profile of CD4+ and CD4- PBMC derived NKT cell lines from RA patients revealed a significantly reduced number of IL-4 producing cells. In contrast, synovial fluid derived NKT cell lines exhibited a Th0-like phenotype, which was comparable to that in healthy control individuals. This suggests that synovial fluid NKT cells are functional, even in patients with nonresponding NKT cells in their blood. We conclude that, because the number of Valpha24+Vbeta11+CD3+ NKT cells is decreased and the cytokine profile of blood-derived NKT cells is biased toward a Th1-like phenotype in RA patients, NKT cells might be functionally related to resistance or progression of RA. Providing a local boost to the regulatory potential of NKT cells might represent a useful candidate therapy for RA. PMID- 15899037 TI - Histone deacetylase inhibitors modulate metalloproteinase gene expression in chondrocytes and block cartilage resorption. AB - Cartilage destruction in the arthritides is thought to be mediated by two main enzyme families: the matrix metalloproteinases (MMPs) are responsible for cartilage collagen breakdown, and enzymes from the ADAMTS (a disintegrin and metalloproteinase domain with thrombospondin motifs) family mediate cartilage aggrecan loss. Many genes subject to transcriptional control are regulated, at least in part, by modifications to chromatin, including acetylation of histones. The aim of this study was to examine the impact of histone deacetylase (HDAC) inhibitors on the expression of metalloproteinase genes in chondrocytes and to explore the potential of these inhibitors as chondroprotective agents. The effects of HDAC inhibitors on cartilage degradation were assessed using a bovine nasal cartilage explant assay. The expression and activity of metalloproteinases was measured using real-time RT-PCR, western blot, gelatin zymography, and collagenase activity assays using both SW1353 chondrosarcoma cells and primary human chondrocytes. The HDAC inhibitors trichostatin A and sodium butyrate potently inhibit cartilage degradation in an explant assay. These compounds decrease the level of collagenolytic enzymes in explant-conditioned culture medium and also the activation of these enzymes. In cell culture, these effects are explained by the ability of HDAC inhibitors to block the induction of key MMPs (e.g. MMP-1 and MMP-13) by proinflammatory cytokines at both the mRNA and protein levels. The induction of aggrecan-degrading enzymes (e.g. ADAMTS4, ADAMTS5, and ADAMTS9) is also inhibited at the mRNA level. HDAC inhibitors may therefore be novel chondroprotective therapeutic agents in arthritis by virtue of their ability to inhibit the expression of destructive metalloproteinases by chondrocytes. PMID- 15899038 TI - ANKH variants associated with ankylosing spondylitis: gender differences. AB - The ank (progressive ankylosis) mutant mouse, which has a nonsense mutation in exon 12 of the inorganic pyrophosphate regulator gene (ank), exhibits aberrant joint ankylosis similar to human ankylosing spondylitis (AS). We previously performed family-based association analyses of 124 Caucasian AS families and showed that novel genetic markers in the 5' flanking region of ANKH (the human homolog of the murine ank gene) are modestly associated with AS. The objective of the present study was to conduct a more extensive evaluation of ANKH variants that are significantly associated with AS and to determine whether the association is gender specific. We genotyped 201 multiplex AS families with nine ANKH intragenetic and two flanking microsatellite markers, and performed family based association analyses. We showed that ANKH variants located in two different regions of the ANKH gene were associated with AS. Results of haplotype analyses indicated that, after Bonferroni correction, the haplotype combination of rs26307 [C] and rs27356 [C] is significantly associated with AS in men (recessive/dominant model; P = 0.004), and the haplotype combination of rs28006 [C] and rs25957 [C] is significantly associated with AS in women (recessive/dominant model; P = 0.004). A test of interaction identified rs26307 (i.e. the region that was associated in men with AS) as showing a difference in the strength of the association by gender. The region associated with AS in women only showed significance in the test of interaction among the subset of families with affected individuals of both genders. These findings support the concept that ANKH plays a role in genetic susceptibility to AS and reveals a gender genotype specificity in this interaction. PMID- 15899039 TI - The mechanism of low-concentration sodium nitroprusside-mediated protection of chondrocyte death. AB - Sodium nitroprusside (SNP), a widely used nitric oxide donor, has recently been shown to mediate chondrocyte apoptosis by generating reactive oxygen species, whereas more potent nitric oxide donors do not induce chondrocyte apoptosis. The present study was performed to investigate the protective effect of a low concentration of SNP upon the cytotoxicity of chondrocytes to higher concentrations of SNP, and to elucidate the underlying mechanism. Human osteoarthritis chondrocytes were cultured as monolayers, and first-passage cells were used for the experiments. Chondrocyte death induced by 1 mM SNP was completely inhibited by pretreating with 0.1 mM SNP. This protective effect of SNP was replicated by the guanosine-3',5'kappa-cyclic monophosphate analog, DBcGMP. Protection from chondrocyte death conferred by 0.1 mM SNP was mediated by heme oxygenase 1 (HO-1), as was revealed by the increased expression of HO-1 in 0.1 mM SNP pretreated chondrocytes and by the reversal of this protective effect by the HO-1 inhibitor, zinc protoporphyrin. SNP-mediated chondrocyte protection correlated with the downregulation of both extracellular signal-regulated protein kinase 1/2 and p38 kinase activation. SNP at 0.1 mM induced significant NF-kappaB activation as revealed by electrophoretic mobility shift assays, and the inhibition of NF-kappaB by MG132 or Bay 11-7082 nullified 0.1 mM SNP-mediated chondrocyte protection. The upregulation of p53 and the downregulation of Bcl-XL and Mcl-1 by 1 mM SNP were reversed by 0.1 mM SNP pretreatment at the protein level by western blotting. Our study shows that priming with 0.1 mM SNP confers complete protection against cell death induced by 1 mM SNP in human articular chondrocytes. This protective effect was found to be correlated with the upregulation of both HO-1 and NF-kappaB and with the concomitant downregulation of both extracellular signal-regulated protein kinase 1/2 and p38 activation. PMID- 15899040 TI - Synovial microparticles from arthritic patients modulate chemokine and cytokine release by synoviocytes. AB - Synovial fluid from patients with various arthritides contains procoagulant, cell derived microparticles. Here we studied whether synovial microparticles modulate the release of chemokines and cytokines by fibroblast-like synoviocytes (FLS). Microparticles, isolated from the synovial fluid of rheumatoid arthritis (RA) and arthritis control (AC) patients (n = 8 and n = 3, respectively), were identified and quantified by flow cytometry. Simultaneously, arthroscopically guided synovial biopsies were taken from the same knee joint as the synovial fluid. FLS were isolated, cultured, and incubated for 24 hours in the absence or presence of autologous microparticles. Subsequently, cell-free culture supernatants were collected and concentrations of monocyte chemoattractant protein-1 (MCP-1), IL-6, IL-8, granulocyte/macrophage colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) and intracellular adhesion molecule-1 (ICAM-1) were determined. Results were consistent with previous observations: synovial fluid from all RA as well as AC patients contained microparticles of monocytic and granulocytic origin. Incubation with autologous microparticles increased the levels of MCP-1, IL-8 and RANTES in 6 of 11 cultures of FLS, and IL-6, ICAM-1 and VEGF in 10 cultures. Total numbers of microparticles were correlated with the IL 8 (r = 0.91, P < 0.0001) and MCP-1 concentrations (r = 0.81, P < 0.0001), as did the numbers of granulocyte-derived microparticles (r = 0.89, P < 0.0001 and r = 0.93, P < 0.0001, respectively). In contrast, GM-CSF levels were decreased. These results demonstrate that microparticles might modulate the release of chemokines and cytokines by FLS and might therefore have a function in synovial inflammation and angiogenesis. PMID- 15899041 TI - Systemic lupus erythematosus induced by anti-tumour necrosis factor alpha therapy: a French national survey. AB - The development of drug-induced lupus remains a matter of concern in patients treated with anti-tumour necrosis factor (TNF) alpha. The incidence of such adverse effects is unknown. We undertook a retrospective national study to analyse such patients. Between June and October 2003, 866 rheumatology and internal medicine practitioners from all French hospital centres prescribing anti TNF in rheumatic diseases registered on the website of the 'Club Rhumatismes et Inflammation' were contacted by email to obtain the files of patients with TNF induced systemic lupus erythematosus. Twenty-two cases were collected, revealing two aspects of these manifestations. Ten patients (six patients receiving infliximab, four patients receiving etanercept) only had anti-DNA antibodies and skin manifestations one could classify as 'limited skin lupus' or 'toxidermia' in a context of autoimmunity, whereas 12 patients (nine patients receiving infliximab, three patients receiving etanercept) had more complete drug-induced lupus with systemic manifestations and at least four American Congress of Rheumatology criteria. One patient had central nervous system manifestations. No patients had lupus nephritis. The signs of lupus occurred within a mean of 9 months (range 3-16 months) in patients treated with infliximab and within a mean of 4 months (range 2-5 months) in patients treated with etanercept. In all cases after diagnosis was determined, anti-TNF was stopped and specific treatment introduced in eight patients: two patients received intravenous methylprednisolone, four patients received oral steroids (15-35 mg/day), and two patients received topical steroids. Lupus manifestations abated within a few weeks (median 8 weeks, standard deviation 3-16) in all patients except one with longer-lasting evolution (6 months). At that time, cautious estimations (unpublished data from Schering Plough Inc. and Wyeth Inc.) indicated that about 7700 patients had been exposed to infliximab and 3000 to etanercept for inflammatory arthritides in France. It thus appears that no drug was more implicated than the other in lupus syndromes, whose incidence was 15/7700 = 0.19% with infliximab and 7/3800 = 0.18% with etanercept. Clinicians should be aware that lupus syndromes with systemic manifestations may occur in patients under anti-TNF alpha treatment. PMID- 15899042 TI - Quantitative ultrasound can assess the regeneration process of tissue-engineered cartilage using a complex between adherent bone marrow cells and a three dimensional scaffold. AB - Articular cartilage (hyaline cartilage) defects resulting from traumatic injury or degenerative joint disease do not repair themselves spontaneously. Therefore, such defects may require novel regenerative strategies to restore biologically and biomechanically functional tissue. Recently, tissue engineering using a complex of cells and scaffold has emerged as a new approach for repairing cartilage defects and restoring cartilage function. With the advent of this new technology, accurate methods for evaluating articular cartilage have become important. In particular, in vivo evaluation is essential for determining the best treatment. However, without a biopsy, which causes damage, articular cartilage cannot be accurately evaluated in a clinical context. We have developed a novel system for evaluating articular cartilage, in which the acoustic properties of the cartilage are measured by introducing an ultrasonic probe during arthroscopy of the knee joint. The purpose of the current study was to determine the efficacy of this ultrasound system for evaluating tissue-engineered cartilage in an experimental model involving implantation of a cell/scaffold complex into rabbit knee joint defects. Ultrasonic echoes from the articular cartilage were converted into a wavelet map by wavelet transformation. On the wavelet map, the percentage maximum magnitude (the maximum magnitude of the measurement area of the operated knee divided by that of the intact cartilage of the opposite, nonoperated knee; %MM) was used as a quantitative index of cartilage regeneration. Using this index, the tissue-engineered cartilage was examined to elucidate the relations between ultrasonic analysis and biochemical and histological analyses. The %MM increased over the time course of the implant and all the hyaline-like cartilage samples from the histological findings had a high %MM. Correlations were observed between the %MM and the semiquantitative histologic grading scale scores from the histological findings. In the biochemical findings, the chondroitin sulfate content increased over the time course of the implant, whereas the hydroxyproline content remained constant. The chondroitin sulfate content showed a similarity to the results of the %MM values. Ultrasonic measurements were found to predict the regeneration process of the tissue-engineered cartilage as a minimally invasive method. Therefore, ultrasonic evaluation using a wavelet map can support the evaluation of tissue-engineered cartilage using cell/scaffold complexes. PMID- 15899043 TI - Proliferation and differentiation potential of chondrocytes from osteoarthritic patients. AB - Autologous chondrocyte transplantation (ACT) has been shown, in long-term follow up studies, to be a promising treatment for the repair of isolated cartilage lesions. The method is based on an implantation of in vitro expanded chondrocytes originating from a small cartilage biopsy harvested from a non-weight-bearing area within the joint. In patients with osteoarthritis (OA), there is a need for the resurfacing of large areas, which could potentially be made by using a scaffold in combination with culture-expanded cells. As a first step towards a cell-based therapy for OA, we therefore investigated the expansion and redifferentiation potential in vitro of chondrocytes isolated from patients undergoing total knee replacement. The results demonstrate that OA chondrocytes have a good proliferation potential and are able to redifferentiate in a three dimensional pellet model. During the redifferentiation, the OA cells expressed increasing amounts of DNA and proteoglycans, and at day 14 the cells from all donors contained type II collagen-rich matrix. The accumulation of proteoglycans was in comparable amounts to those from ACT donors, whereas total collagen was significantly lower in all of the redifferentiated OA chondrocytes. When the OA chondrocytes were loaded into a scaffold based on hyaluronic acid, they bound to the scaffold and produced cartilage-specific matrix proteins. Thus, autologous chondrocytes are a potential source for the biological treatment of OA patients but the limited collagen synthesis of the OA chondrocytes needs to be further explained. PMID- 15899044 TI - Synovial histopathology of psoriatic arthritis, both oligo- and polyarticular, resembles spondyloarthropathy more than it does rheumatoid arthritis. AB - At present only few biological data are available to indicate whether psoriatic arthritis (PsA) is part of the spondyloarthropathy (SpA) concept, whether it is a separate disease entity or a heterogeneous disease group with oligoarticular/axial forms belonging to SpA and polyarticular forms resembling rheumatoid arthritis (RA). To address this issue with regard to peripheral synovitis, we compared the synovial characteristics of PsA with those of ankylosing spondylitis (AS)/undifferentiated SpA (USpA) and RA, and compared the synovium of oligoarticular versus polyarticular PsA. Synovial biopsies were obtained from patients with RA, nonpsoriatic SpA (AS + USpA), and oligoarticular and polyarticular PsA. The histological analysis included examination(s) of the lining layer thickness, vascularity, cellular infiltration, lymphoid aggregates, plasma cells and neutrophils. Also, we performed immunohistochemical assessments of CD3, CD4, CD8, CD20, CD38, CD138, CD68, CD163, CD83, CD1a, CD146, alphaVbeta3, E-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, S100A12, intracellular citrullinated proteins and major histocompatibility complex (MHC)-human cartilage (HC) gp39 peptide complexes. Comparing SpA (PsA + AS + USpA) with RA, vascularity, and neutrophil and CD163+ macrophage counts were greater in SpA (P < 0.05), whereas lining layer thickness and the number of CD83+ dendritic cells were greater in RA (P < 0.05). In RA, 44% of samples exhibited positive staining for intracellular citrullinated proteins and 46% for MHC-HC gp39 peptide complexes, whereas no staining for these markers was observed in SpA samples. We excluded influences of disease-modifying antirheumatic drug and/or corticosteroid treatment by conducting systematic analyses of treated and untreated subgroups. Focusing on PsA, no significant differences were observed between PsA and nonpsoriatic SpA. In contrast, vascularity (P < 0.001) and neutrophils were increased in PsA as compared with RA (P = 0.010), whereas staining for intracellular citrullinated proteins and MHC-HC gp39 peptide complexes was exclusively observed in RA (both P = 0.001), indicating that the same discriminating features are found in PsA and other SpA subtypes compared with RA. Exploring synovial histopathology between oligoarticular and polyarticular PsA, no significant differences were noted. Moreover, intracellular citrullinated proteins and MHC-HC gp39 peptide complexes, which are specific markers for RA, were observed in neither oligoarticular nor polyarticular PsA. Taken together, these data indicate that the synovial histopathology of PsA, either oligoarticular or polyarticular, resembles that of other SpA subtypes, whereas both groups can be differentiated from RA on the basis of these same synovial features, suggesting that peripheral synovitis in PsA belongs to the SpA concept. PMID- 15899045 TI - Signalling pathway involved in nitric oxide synthase type II activation in chondrocytes: synergistic effect of leptin with interleukin-1. AB - The objective of the present study was to investigate the effect of leptin, alone or in combination with IL-1, on nitric oxide synthase (NOS) type II activity in vitro in human primary chondrocytes, in the mouse chondrogenic ATDC5 cell line, and in mature and hypertrophic ATDC5 differentiated chondrocytes. For completeness, we also investigated the signalling pathway of the putative synergism between leptin and IL-1. For this purpose, nitric oxide production was evaluated using the Griess colorimetric reaction in culture medium of cells stimulated over 48 hours with leptin (800 nmol/l) and IL-1 (0.025 ng/ml), alone or combined. Specific pharmacological inhibitors of NOS type II (aminoguanidine [1 mmol/l]), janus kinase (JAK)2 (tyrphostin AG490 and Tkip), phosphatidylinositol 3-kinase (PI3K; wortmannin [1, 2.5, 5 and 10 micromol/l] and LY294002 [1, 2.5, 5 and 10 micromol/l]), mitogen-activated protein kinase kinase (MEK)1 (PD098059 [1, 5, 10, 20 and 30 micromol/l]) and p38 kinase (SB203580 [1, 5, 10, 20 and 30 micromol/l]) were added 1 hour before stimulation. Nitric oxide synthase type II mRNA expression in ATDC5 chondrocytes was investigated by real time PCR and NOS II protein expression was analyzed by western blot. Our results indicate that stimulation of chondrocytes with IL-1 results in dose-dependent nitric oxide production. In contrast, leptin alone was unable to induce nitric oxide production or expression of NOS type II mRNA or its protein. However, co stimulation with leptin and IL-1 resulted in a net increase in nitric oxide concentration over IL-1 challenge that was eliminated by pretreatment with the NOS II specific inhibitor aminoguanidine. Pretreatment with tyrphostin AG490 and Tkip (a SOCS-1 mimetic peptide that inhibits JAK2) blocked nitric oxide production induced by leptin/IL-1. Finally, wortmannin, LY294002, PD098059 and SB203580 significantly decreased nitric oxide production. These findings were confirmed in mature and hypertrophic ATDC5 chondrocytes, and in human primary chondrocytes. This study indicates that leptin plays a proinflammatory role, in synergy with IL-1, by inducing NOS type II through a signalling pathway that involves JAK2, PI3K, MEK-1 and p38 kinase. PMID- 15899046 TI - Anti-Sa antibodies and antibodies against cyclic citrullinated peptide are not equivalent as predictors of severe outcomes in patients with recent-onset polyarthritis. AB - The prognostic value of two antibodies targeting citrullinated antigens, anti-Sa and anti-cyclic citrullinated peptide (CCP), present at inclusion, was evaluated prospectively in a cohort of 165 consecutive patients with recent-onset or early polyarthritis (EPA) followed for up to 30 months. Patients were treated according to current Good Clinical Practice standards. Predefined outcomes were severe arthritis and persistent arthritis. At inclusion, a median of 3 months after disease onset, 133 (81%) patients fulfilled at least four American College of Rheumatology criteria for rheumatoid arthritis and 30 (18%) had erosive changes on radiographs of hands and feet. Disease-modifying anti-rheumatic drugs were used in close to 80% of the patients at 30 months. Joint damage increased linearly over time, whereas disease activity declined markedly and remained low at each follow-up. Autoantibodies were identified in 76 (46%) patients: rheumatoid factor (RF) in 68 (41%), anti-CCP in 53 (33%), and anti-Sa in 46 (28%). All three antibodies were correlated, but anti-Sa antibodies best predicted severity at 18 and 30 months. RF and anti-CCP performed less well. For both outcomes, anti-Sa alone performed better than any combination of antibodies. The presence of any autoantibody identified about 50 to 60% of the patients with poor outcomes. In multivariate analysis, anti-Sa (odds ratio (OR) 8.83), the presence of erosions at inclusion (OR 3.47) and increasing age (OR 1.06/year) were significantly associated with severity, whereas RF and anti-CCP were not significant predictors. Persistent arthritis was present in up to 84% of patients; autoantibodies were specific but poorly sensitive predictors of this outcome. We conclude that assays for antibodies against citrullinated antigens differ in their ability to predict poorer outcomes in patients with EPA. In our EPA cohort treated in accordance with current standards, detection of anti-Sa but not of RF or anti-CCP antibodies, in combination with clinical and radiological variables present at the first encounter, allowed the identification of a subgroup of EPA patients suffering more rapid and more severe joint damage over 30 months. PMID- 15899047 TI - CD134 as target for specific drug delivery to auto-aggressive CD4+ T cells in adjuvant arthritis. AB - T cells have an important role during the development of autoimmune diseases. In adjuvant arthritis, a model for rheumatoid arthritis, we found that the percentage of CD4+ T cells expressing the activation marker CD134 (OX40 antigen) was elevated before disease onset. Moreover, these CD134+ T cells showed a specific proliferative response to the disease-associated epitope of mycobacterial heat shock protein 60, indicating that this subset contains auto aggressive T cells. We studied the usefulness of CD134 as a molecular target for immune intervention in arthritis by using liposomes coated with a CD134-directed monoclonal antibody as a drug targeting system. Injection of anti-CD134 liposomes subcutaneously in the hind paws of pre-arthritic rats resulted in targeting of the majority of CD4+CD134+ T cells in the popliteal lymph nodes. Furthermore, we showed that anti-CD134 liposomes bound to activated T cells were not internalized. However, drug delivery by these liposomes could be established by loading anti-CD134 liposomes with the dipalmitate-derivatized cytostatic agent 5' fluorodeoxyuridine. These liposomes specifically inhibited the proliferation of activated CD134+ T cells in vitro, and treatment with anti-CD134 liposomes containing 5'-fluorodeoxyuridine resulted in the amelioration of adjuvant arthritis. Thus, CD134 can be used as a marker for auto-aggressive CD4+ T cells early in arthritis, and specific liposomal targeting of drugs to these cells via CD134 can be employed to downregulate disease development. PMID- 15899048 TI - Increased serum HO-1 in hemophagocytic syndrome and adult-onset Still's disease: use in the differential diagnosis of hyperferritinemia. AB - Heme oxygenase-1 (HO-1), an inducible heme-degrading enzyme, is expressed by macrophages and endothelial cells in response to various stresses. Because ferritin synthesis is stimulated by Fe2+, which is a product of heme degradation, we examined the relation between HO-1 and ferritin levels in the serum of patients with hemophagocytic syndrome (HPS), adult-onset Still's disease (ASD), and other diseases that may cause hyperferritinemia. Seven patients with HPS, 10 with ASD, 73 with other rheumatic diseases, 20 with liver diseases, 10 recipients of repeated blood transfusion because of hematological disorders, and 22 healthy volunteers were enrolled. Serum HO-1 and ferritin levels were determined by ELISA. Expression of HO-1 mRNA and protein by peripheral blood mononuclear cells (PBMCs) was determined by real-time PCR and immunocytochemical techniques, respectively. Serum levels of HO-1 were significantly higher in patients with active HPS and ASD than in the other groups (P < 0.01). HO-1 levels were not elevated in patients with other causes of hyperferritinemia but were moderately elevated in patients with dermatomyositis/polymyositis. Among patients with HPS and ASD, serum HO-1 levels correlated closely with serum ferritin levels, and the levels of both returned to normal after therapy had induced remission. Increased expression of HO-1 mRNA was confirmed in PBMCs from some patients with HPS and ASD. Hyperferritinemia correlated closely with increased serum HO-1 in patients with HPS and ASD but not other conditions, indicating that measurement of serum HO-1 and ferritin levels would be useful in the differential diagnosis of hyperferritinemia and perhaps also in monitoring disease activity in HPS and ASD. PMID- 15899049 TI - Effect of risedronate on joint structure and symptoms of knee osteoarthritis: results of the BRISK randomized, controlled trial [ISRCTN01928173]. AB - To determine the efficacy and safety of risedronate in patients with knee osteoarthritis (OA), the British study of risedronate in structure and symptoms of knee OA (BRISK), a 1-year prospective, double-blind, placebo-controlled study, enrolled patients (40-80 years of age) with mild to moderate OA of the medial compartment of the knee. The primary aims were to detect differences in symptoms and function. Patients were randomized to once-daily risedronate (5 mg or 15 mg) or placebo. Radiographs were taken at baseline and 1 year for assessment of joint space width using a standardized radiographic method with fluoroscopic positioning of the joint. Pain, function, and stiffness were assessed using the Western Ontario and McMaster Universities (WOMAC) OA index. The patient global assessment and use of walking aids were measured and bone and cartilage markers were assessed. The intention-to-treat population consisted of 284 patients. Those receiving risedronate at 15 mg showed improvement of the WOMAC index, particularly of physical function, significant improvement of the patient global assessment (P < 0.001), and decreased use of walking aids relative to patients receiving the placebo (P = 0.009). A trend towards attenuation of joint-space narrowing was observed in the group receiving 15 mg risedronate. Eight percent (n = 7) of patients receiving placebo and 4% (n = 4) of patients receiving 5 mg risedronate exhibited detectable progression of disease (joint-space width >or= 25% or >or= 0.75 mm) versus 1% (n = 1) of patients receiving 15 mg risedronate (P = 0.067). Risedronate (15 mg) significantly reduced markers of cartilage degradation and bone resorption. Both doses of risedronate were well tolerated. In this study, clear trends towards improvement were observed in both joint structure and symptoms in patients with primary knee OA treated with risedronate. PMID- 15899050 TI - Biomarkers of endothelial dysfunction, cardiovascular risk factors and atherosclerosis in rheumatoid arthritis. AB - Cardiovascular event rates are markedly increased in rheumatoid arthritis (RA), and RA atherogenesis remains poorly understood. The relative contributions of traditional and nontraditional risk factors to cardiovascular disease in RA await elucidation. The present study comprises three components. First, we compared biomarkers of endothelial dysfunction (vascular cell adhesion molecule [VCAM]-1, intercellular adhesion molecule [ICAM]-1 and endothelial leucocyte adhesion molecule [ELAM]-1) in 74 RA patients and 80 healthy control individuals before and after controlling for traditional and nontraditional cardiovascular risk factors, including high-sensitivity C-reactive protein (hs-CRP), IL-1, IL-6 and tumor necrosis factor-alpha. Second, we investigated the potential role of an extensive range of patient characteristics in endothelial dysfunction in the 74 RA patients. Finally, we assessed associations between biomarkers of endothelial dysfunction and ultrasonographically determined common carotid artery intima media thickness and plaque in RA. The three biomarkers of endothelial dysfunction, as well as hs-CRP, IL-1, IL-6 and tumor necrosis factor-alpha, were higher in patients than in control individuals (P < 0.0001). Patients were also older, exercised less and had a greater waist circumference, blood pressure and triglyceride levels (P /=1 model, only 1.4% met criteria for all 3. CONCLUSIONS: Application of available models to a screening population classifies a larger than expected number of women at high risk for a BRCA1 or BRCA2 mutation. New approaches to risk assessment and counseling are needed to apply our knowledge of hereditary risk to a broad population in a practical manner. PMID- 15899072 TI - Patient and physician attitudes toward breast cancer clinical trials: developing interventions based on understanding barriers. AB - PURPOSE: Clinical trials are essential to develop and test novel therapies, yet only 2%-3% of women with breast cancer enroll. We sought to describe patient and physician barriers to trial participation and then implemented targeted interventions to increase awareness and interest in trial participation. Also, with increasing patient interest in complementary and alternative medicine (CAM) for cancer, we explored attitudes regarding CAM clinical trials. PATIENTS AND METHODS: Between 1997 and 2000, questionnaires were offered to patients with newly diagnosed or recurrent breast cancer and to physicians specializing in breast cancer. Programs aimed at patients and physicians from our geographic region to increase their support for breast cancer clinical trials were initiated in 1997. Correlation between perceived barriers and patient and physician demographics were explored. Reluctance to be randomized, extra time, and concerns about worse side effects with the experimental arm were the most significant patient barriers. Physician barriers included randomization, extra staff time, and increased costs of enrollment. Patients and physicians approved of studying CAM in clinical trials, with different scores based on age and type of practice. Physicians and patients developed more favorable views of clinical trials between 1997 and 2000. RESULTS: Although many barriers still exist, this study suggests that attitudes toward clinical trials are evolving and significantly affected by patient age and stage of disease. Because different patients and some different physicians were surveyed, it is difficult to conclude that the changes occurred as a result of the interventions. CONCLUSION: Future efforts to improve enrollment should focus on patients' individual concerns and the uneasiness with the randomization. PMID- 15899073 TI - A phase II trial of gemcitabine in patients with metastatic breast cancer previously treated with an anthracycline and taxane. AB - PURPOSE: This study was designed to evaluate the efficacy and safety of single agent gemcitabine for the treatment of patients with anthracycline- and taxane pretreated metastatic breast cancer (MBC). Eligible patients were required to have bidimensionally measurable MBC that had been treated with 2-4 prior chemotherapy regimens that included an anthracycline and a taxane. Gemcitabine was delivered at a dose of 800 mg/m2 on days 1, 8, and 15 of a 28-day cycle until evidence of disease progression. PATIENTS AND METHODS: Twenty-two patients were enrolled and included in the safety analysis; 18 patients were evaluable for response. The median age of patients was 54 years (range, 36-70 years). The mean number of prior chemotherapy regimens for metastatic disease was 2.3, and the mean dose of gemcitabine delivered was 911 mg/m2 (range, 600-1600 mg/m2). RESULTS: Overall, gemcitabine was well tolerated with minimal grade 3 toxicities; the only grade 4 toxicity was 1 case of pulmonary embolus. Three patients had evidence of partial tumor regression (17%; 95% CI, 4%-41%), and 1 patient had a 41% decrease in tumor volume, including liver metastasis. CONCLUSIONS: Gemcitabine is active and well tolerated as monotherapy given in heavily pretreated patients with MBC after anthracyclines and taxanes. The activity and safety reported in this trial are consistent with previous reports in similar patients. PMID- 15899074 TI - Bringing molecular prognosis and prediction to the clinic. AB - In the past 30 years, important advances have been made in the knowledge of breast cancer biology and in the treatment of the disease. However, the translation of these advances into clinical practice has been slow. With the advent of molecular-based medicine, it is hoped that the bridge between the bench and the bedside will continue to be shortened. Because breast cancer is a heterogeneous disease with wide-ranging subsets of patients who have different prognoses and who respond differently to treatments, the identification of patients who need treatment and the definition of the best therapy for an individual have become the priorities in breast cancer care. This article will review the crucial role of prognostic and predictive factors in achieving these goals. A critical review of classical and newer individual molecular markers, such as hormone receptors, HER2, urokinase-type plasminogen activator and plasminogen activator inhibitor 1, cyclin E, topoisomerase II, and p53, was performed, and the preliminary results obtained using the new gene expression profiling technology are described along with their potential clinical implications. PMID- 15899075 TI - Incorporating trastuzumab into the neoadjuvant treatment of HER2-overexpressing breast cancer. AB - We sought to provide a brief overview of the available results of trastuzumab in the neoadjuvant treatment of women with locally advanced, HER2-overexpressing breast cancer. A review of published reports was conducted by the use of Medline computer searching and manual searching of abstracts presented at the Annual Meeting of the American Society of Clinical Oncology and the San Antonio Breast Cancer Symposium between 2002 and 2004. Several phase II trials and one phase III trial show that trastuzumab-containing regimens are feasible and yield high rates of clinical and pathologic complete response in women with locally advanced HER2 overexpressing breast cancer. Moreover, the neoadjuvant approach allows serial monitoring of biologic phenomena that occur during the administration of therapy, which acquires particular importance in the case of targeted therapies like trastuzumab. The incorporation of trastuzumab into neoadjuvant therapy for breast cancer appears promising. However, no data on long-term survival and safety are available at present. Well-designed clinical trials are needed to establish the optimal trastuzumab-based neoadjuvant strategy. PMID- 15899076 TI - Should we still be testing for asymptomatic non-specific urethritis in departments of genitourinary medicine? AB - It has recently been advocated that non-invasive testing with first-catch urine specimens using nucleic acid amplification techniques, to detect Chlamydia trachomatis and Neisseria gonorrhoeae, should replace routine microscopy on asymptomatic men. Although it is assumed that this strategy will be cost effective, the available evidence suggests that this will result in fewer sexually transmitted infections being averted than continuing the current practice of screening for urethritis and testing for both microorganisms in asymptomatic men. This review article summarizes the available evidence and argues that research is urgently needed in order to properly evaluate the cost effectiveness of detecting urethritis in asymptomatic men. PMID- 15899077 TI - Introduction of a proforma in the management of under age attendees at a genitourinary clinic. AB - The aim of this review was evaluation of a recently developed proforma for improving assessment and management of patients under the age of 16 in the genitourinary (GU) medicine clinic. A case-note review of all under-16s attending between June 2000 and March 2001 was undertaken (109 patients). Comparison with review from 1998 prior to proforma introduction was carried out. In all, 99 proformas were completed. Fewer young patients were seen solely by junior doctors since proforma introduction (27-45%) (P=0.012), more were referred to health advisors (79% versus 66%) (P = 0.056),but follow-up remains suboptimal (72% versus 78%). Possibility of abuse was assessed in 102 patients (17 cases of non consensual sex versus six in 1998). In all, 54% were using no contraception and only 21% were consistently using barrier methods; 41% had sexually transmitted infections diagnosed. The proforma is useful for collecting data and directing management when completed fully, and has revealed greater numbers of children involved in risky behaviour and abuse. PMID- 15899078 TI - The role of the nurse in screening asymptomatic male and female patients in a sexual health clinic. AB - We allocated 278 patients, who said they were asymptomatic and agreed to be randomized to a nurse or doctor clinic, to appointments using a random number system. In all, 35 patients did not attend and 16 were excluded because they did not meet the entry criteria. We used a screening protocol which excluded microscopy from the immediate assessment of patients. The outcome measures were completeness of documentation, proportion of patients accepting HIV tests, infections detected and patient satisfaction. Overall, 3% of items were not completed by doctors and 6% by nurses. HIV tests were carried out on 65% of patients who saw a doctor and 52% who saw a nurse. Thirteen infections were detected by doctors and 27 by nurses. No new cases of gonorrhoea, syphilis or HIV infection were identified. Eighty-eight patients completed a questionnaire after their attendance. Almost all patients were very satisfied with the service and most were prepared to see a nurse on a subsequent visit. We concluded that there are few differences between the performance of doctors and nurses in routine screening of asymptomatic patients. PMID- 15899079 TI - Treatment failure of vaginal trichomoniasis in clinical practice. AB - We have conducted a retrospective study of treatment failure of Trichomonas vaginalis (TV) infection in a busy, urban genitourinary medicine clinic in Birmingham, UK, between 1998 and 2002. The prevalence of non-response to standard doses of metronidazole without any history of reinfection or nonadherence was 1.7% (28/1603) overall, and has significantly increased from 0.38% in 1999 to 3.5% in 2002 (P = 0.001, chi2 test). There were no associated demographic factors. The majority of patients (53% or 15/28) responded to either a repeat of standard (10/26) or higher oral dose (5/7) of metronidazole. A standardized treatment algorithm using a high dose of metronidazole may be useful in the absence of any new, efficacious non-imidazole-based TV therapy in the clinical management of persistent TV infection. PMID- 15899080 TI - Treatment and partner notification outcomes for gonorrhoea: effect of ethnicity and gender. AB - We assessed if gender and ethnicity were associated with differences in the number of patients satisfactorily treated and number of partners successfully treated for genital gonococcal infection by reviewing 400 cases. There were no differences in the rates of satisfactory treatment of gonorrhoea between men and women and the different ethnic groups. There was no difference in satisfactory partner treatment rates by ethnic origin; however, men were less likely to have their contacts satisfactorily treated within 28 days of diagnosis (odds ratio = 0.05, 95% confidence interval 0.3-0.7). This inequality may lead to difficulties in reducing the number of new gonorrhoea cases. More research is needed to find out why men behave differently from women regarding partner notification for genital gonorrhoea infection. PMID- 15899081 TI - Higher vaginal pH is associated with Chlamydia trachomatis infection in women: a prospective case-controlled study. AB - This is a prospective case-controlled study of female attendees in Coventry. This study found an association of higher vaginal pH with chlamydial infection, independent of any other factors. Studies in vitro have shown that an acidic vaginal secretion inhibits chlamydial infection. Our objective was to analyse the association of vaginal pH and chlamydial infection in women attending a genitourinary medicine clinic. Chlamydial infections were diagnosed with ELISA and confirmed within direct immunofluorescence. Vaginal pH was measured by a pH indicator tape ranging from 3 to 8. Consecutive female attendees with no sexually transmitted infections (STIs) were included as controls. In all, 144 female cases, diagnosed with chlamydial infection, had a median age of 20 years. Seventeen women had associated bacterial vaginosis. Eighty-two women had no other STIs. Ninety-eight women were using the oral contraceptive pill (OCP). The 145 control women had a median age of 26 years and 52 were receiving the OCP. A significantly higher vaginal pH was seen in the cases (P = 0.0001, Wilcoxon test), even after adjusting for other risk factors associated with vaginal pH, including OCP use (odds ratio: 6.49, 95% confidence interval, 3.59-11.73, P = 0.0001). Chlamydial infection in women was associated with a higher vaginal pH level, independent of any other factors. This study has implications for the treatment of other conditions known to lead to an increase in vaginal pH, even in asymptomatic individuals. PMID- 15899082 TI - Evaluation of the serodia Treponema pallidum particle agglutination, the Murex Syphilis ICE and the Enzywell TP tests for serodiagnosis of syphilis. AB - We evaluated the Treponema pallidum haemagglutination assay (TPHA), a treponemal test, with three other treponemal tests, the Serodia T. pallidum particle agglutination assay, the Murex Syphilis ICE IgG + IgM enzyme immunoassay (EIA) and the Enzywell TP IgG + M EIA (a new rapid EIA) for use in conjunction with the rapid plasma reagin test (RPR), a non-treponemal test, for serodiagnosis of syphilis. In all, 124 serum samples were found reactive with RPR and/or TPHA after testing by the routine laboratory protocol. Twenty-three (18.5%) of them were positive only by RPR test and were evaluated as biologically false-positive, 16 were positive only by the TPHA and 84 by both the RPR and TPHA tests; one sample was non-specific (heterophile reaction) in the TPHA. Agreements of the TPHA with the Serodia TPPA, the Murex Syphilis ICE and the Enzywell TP tests were 96.7%, 100% and 99.1%, respectively. We conclude that each one of the tests, the Serodia TPPA, the Murex Syphilis ICE and the Enzywell TP, is an appropriate substitute for screening for serodiagnosis of syphilis. PMID- 15899083 TI - An oral fluid test for syphilis. AB - We have developed a time-resolved fluorescence immunoassay to detect antibodies to Treponema pallidum recombinant antigens in oral fluid specimens. Using an 'Oracol' swab, oral fluid was collected from 34 subjects with a serological diagnosis of syphilis and 97 seronegative controls. Using a cut-off of three standard deviations over control mean, the sensitivity and specificity of the assay in all subjects with positive syphilis serology was 76.5% and 97.9%, respectively. In those with early syphilis, the sensitivity and specificity of the assay was 100% and 97.9%. In a non-outbreak situation, screening clinic attendees for syphilis using oral fluid specimens is potentially useful when collection of blood is not practicable. In addition, it may have much to offer in outreach projects and epidemiological investigations. PMID- 15899084 TI - Prevalence of Toxoplasma gondii infection and incidence of toxoplasma encephalitis in non-haemophiliac HIV-1-infected adults in Taiwan. AB - We assessed the seroprevalence of Toxoplasma gondii infection and incidence of toxoplasma encephalitis (TE) in 844 non-haemophiliac HIV-infected patients in Taiwan between June 1994 and April 2003. Approximately 70% (69.3%) of them had a baseline CD4+ lymphocyte count of 200 x 10(6)/L or less, and more than 70% (73.9%) having initiated highly active antiretroviral therapy. The seroprevalence of T. gondii infection was 10.2%, which did not differ with sex,age,route of transmission, birth inside or outside of Taiwan, or CD4+ lymphocyte stratifications. After a median observation duration of 603 days (range, 1-3264 days), 10 (1.2%) patients developed 11 episodes of TE after a median interval of 30 days (range, 1-941 days) between enrolment and diagnosis of TE, with an incidence of 0.59 per 100 person-years (PY) (95% confidence interval, 0.56-0.63 per 100 PY). We concluded that the incidence of TE of HIV-infected patients in Taiwan was lower than that reported in western countries because of a lower seroprevalence of T. gondii infection and use of antimicrobial prophylaxis and antiretroviral therapy, although most of the patients were at the late stage of HIV infection. PMID- 15899085 TI - Use of bioelectrical impedance analysis to determine body composition changes in HIV-associated wasting. AB - AIDS wasting syndrome results in loss of lean body mass and body cell mass. This 12-week, open-label study used bioelectrical impedance analysis to measure body composition changes in 24 patients with AIDS wasting syndrome receiving recombinant human growth hormone (r-hGH). The primary endpoint was percentage monthly change in body weight before/after r-hGH. Secondary endpoints included change from baseline in body composition (bioelectrical impedance analysis), isometric strength and CD4+ count. Twenty patients completed the study: r-hGH resulted in mean weight gains (+2.7%, P = 0.146), and significant increases in mean body cell mass (+8.0%, P = 0.0211), lean body mass (+4.8%, P = 0.0373) and water (+5.5%, P < 0.023). Body fat decreased throughout, but not significantly. r hGH was generally well tolerated; the most frequent adverse events were fever (7.3%) and diarrhoea (6.3%). Thus, bioelectrical impedance analysis can detect improved body cell mass independent of changes in body weight resulting from r hGH treatment in patients with AIDS wasting syndrome. PMID- 15899086 TI - Willingness to participate in HIV vaccine trials among HIV-negative gay men in Sydney, Australia. AB - This study aimed to determine and describe HIV-negative gay men's willingness to participate in HIV vaccine trials. Data were from participants who completed face to-face interviews during the first 18 months (to the end of 2002) of recruitment into the Health in Men cohort of HIV-negative gay men in Sydney. A key outcome measure was a scale of Willingness to Participate in HIV Vaccine Trials, with scores ranging from 1 (unwilling) to 4 (willing). The 903 participants ranged in age from 18 to 75 years (median = 36). Mean of Willingness to Participate in HIV Vaccine Trials was 2.53 (standard deviation = 0.54), with approximately 51% of the men having a score greater than the midpoint of 2.50. A reduced linear regression model yielded four significant independent associations with Willingness to Participate in HIV Vaccine Trials: lack of tertiary education (P < 0.001),having engaged 'in the previous six months' in any unprotected anal intercourse with casual or non-concordant regular partners (P < 0.001), higher self-rated likelihood of HIV infection (P < 0.01), and higher mean scores on a scale of Comfort with Participation in HIV Vaccine Trials (P < 0.001). The willingness of HIV-negative gay men at potentially higher risk for HIV to participate in HIV vaccine trials augurs well for enrolment in HIV vaccine efficacy trials. Recruitment into trials is likely to be enhanced by addressing salient concerns such as confidentiality and vaccine-induced antibody positivity. Also, it will be important to ensure that gay men are educated and fully informed about HIV vaccines and trial processes. PMID- 15899087 TI - Sociodemography of genital co-infection with Neisseria gonorrhoeae and Chlamydia trachomatis in Coventry, UK. AB - While genital co-infection with Neisseria gonorrhoeae and Chlamydia trachomatis in the same individual is relatively common, little is known about the characteristics of individuals co-infected with both pathogens. We describe the sociodemographic and geographic characteristics of those with genital co infection with N. gonorrhoeae and C. trachomatis.We reviewed the case-notes of all patients presenting with co-infection between March 1989 and February 2000. Incidence rates were calculated for those aged 15-64 living in the 18 different electoral wards of the city and subjects were assigned a Townsend deprivation score based on residence. A total of 332 cases of co-infection were included over the study period (overall mean annual incidence rate 16.1 [95% confidence interval [CI] 9.9-22.3]/100,000). The infection rate was significantly higher in those of black ethnicity (rate: 82.6/10(5), relative rate 5.81, 95% CI [4.03 8.38],P = 0.0001) than in those of other ethnicities. The highest incidence was noted in men aged 20-24 (n = 81, 45.6%) and in women aged 15-19 (n = 66, 45.2%) years, living in the most deprived area of the city. After controlling for year of diagnosis, those aged 25-64 years had significantly lower incidence rates (0.13 [0.10-0.17], P = 0.0001, Poisson regression) than those aged < 20 years. Increased incidence rates were also associated with high deprivation scores. There is a complex interaction between age, sex, ethnicity, geographic distribution, social deprivation and the risk of acquiring genital co-infection with N. gonorrhoeae and C. trachomatis. This study may help to identify the geographic areas of high incidence of sexually transmitted diseases in Coventry, and could be used as the baseline to measure the need for subsequent interventions. PMID- 15899088 TI - Using chlamydia positivity to estimate prevalence: evidence from the Chlamydia Screening Pilot in England. AB - Studies have suggested that positivity can be used to estimate the prevalence of Chlamydia trachomatis in large-scale chlamydia screening programmes. A recent pilot of opportunistic screening in England estimated that the prevalence among 16-24-year-old women in Portsmouth and Wirral was 9.8% and 11.2%, respectively. This study assessed the continued validity of positivity as an approximate for prevalence. We re-analysed data from the Chlamydia Screening Pilot to estimate positivity,calculated as total positive tests divided by total tests, and compared these estimates with the previously reported prevalence, measured as the number of women testing positive divided by the total number of women screened. Overall positivity was 9.4% in Portsmouth and 11.0% in the Wirral; these estimates were not statistically different from prevalence, regardless of health care setting, age group or symptoms. We conclude that positivity can be used as a proxy for prevalence. PMID- 15899089 TI - The national strategy for sexual health and HIV: the achievable progress. PMID- 15899090 TI - General practitioners' involvement with the national strategy on sexual health and HIV. PMID- 15899091 TI - Asymptomatic Chlamydia trachomatis-negative non-gonococcal urethritis. PMID- 15899092 TI - AIDS diagnosed in a middle-aged female psychiatric patient. PMID- 15899093 TI - The forgotten sexual health player: the unacknowledged role of community sexual and reproductive health clinics in managing uncomplicated sexually transmitted infections. PMID- 15899094 TI - Closing the osteoporosis management gap in primary care: a secondary prevention of fracture programme. AB - BACKGROUND: Effective treatments are available to reduce fracture risk in patients with osteoporosis. Prioritisation of assessment and treatment for those patients at highest risk of fracture will and treatment for those patients at highest risk of fracture will will ensure the optimal utilisation of healthcare resources. OBJECTIVES: To confirm prior fracture to be a strong predictor of osteoporosis, evaluate a simple means of identifying patients with osteoporosis, assess the current management gap in this high risk patient group and to enable initiation of treatment where appropriate. RESEARCH DESIGN AND METHODS: All women >/=65 years of age living at home and registered with a general practitioner (GP) in Coatbridge, Lanarkshire, Scotland (4045) were mailed an osteoporosis questionnaire. Participants were from an area of generally low socioeconomic background, where 16% of the population are over >/=65 years and >/=99% are Caucasian. Those who had sustained a fracture or had >/=2 osteoporosis risk factors and had not previously been screened for osteoporosis were invited for a Dual energy X-ray Absorptiometry scan. A second group of women at high risk of osteoporosis were referred by their GP for a scan. Bone mineral density (BMD) was determined and treatment was reviewed and prescribed according to national guidelines. RESULTS: 2386/4045 women returned the questionnaire (response rate 59%); 2286 were correctly completed and made up the sample size. Eight hundred and fifty two had sustained >/=1 fracture(s), of whom 43 (5%) had previously had BMD testing and 80 (9.4%) were receiving treatment. There were 1434 women with no history of fracture that had >/=2 risk factors for osteoporosis. Of 395 women referred by their GP, 113 had sustained fractures. Following the audit, 1054 women were scanned, including 463 women who had not sustained fractures that had >/=2 osteoporosis risk factors. Of the 1054, 591 women had sustained 763 fractures: 46 (6.0%) hip, 284 (37.2%) wrist, 37 (4.8%) humerus and 396 (51.9%) other bones (mainly ankle or rib). Eighty (13.5%) women with a fracture history had normal BMD, 204 (34.5%) were osteopenic and 307 (51.9%) were osteoporotic. Older women were more likely to have osteoporosis: overall, 12.8%, 46.8% and 63.0% of women were osteoporotic in age groups <65 years, 65-75 years and >75 years, respectively. Treatment was prescribed according to Lanarkshire's osteoporosis guidelines for 670 (63.6%) patients: 90.0% received bisphosphonate + calcium/vitamin D and 10% received calcium/vitamin D. CONCLUSIONS: A simple scan identified patients with prior fracture and with osteoporosis. Prior fracture was confirmed to be a strong predictor of osteoporosis; 86.4% of women with a fracture history had low BMD and 51.9% had osteoporosis. Similar disease management programmes elsewhere in primary care to identify high risk patients and ensure appropriate prescribing would, in addition to implementing national guidelines, be pharmaco-economically prudent and improve management of patients with fragility fracture across the UK. PMID- 15899095 TI - Pharmacokinetic simulation for switching from galantamine immediate-release to extended-release formulation. AB - OBJECTIVE: To demonstrate using pharmacokinetic (PK) modeling and simulation, that the PK that the PK parameters for drug exposure with galantamine parameters for drug exposure with galantamine immediate-release (IR) tablet and galantamine extended-release (ER) capsule are comparable in patients with Alzheimer's disease (AD) during in patients with Alzheimer's disease (AD) during the switch from twice-daily IR tablet at steady state to the new once-daily ER capsule, and to support a recommendation that patients receiving the IR tablet at steady state can be successfully switched to the ER capsule at the same daily dosage with no titration period. METHODS: Simulations were performed using a population PK model developed from clinical studies with IR galantamine in the target AD population, in combination with IR and ER absorption parameters obtained from a PK study in healthy volunteers which showed similar results. PK simulations were performed for the switch from IR tablet 8 mg b.i.d. to ER capsule 16 mg q.d. and from IR tablet 12 mg b.i.d. to ER capsule 24 mg q.d. RESULTS: This simulation predicted that patients switched from the IR tablet to the ER capsule, the PK parameters for drug exposure on the first day of ER treatment would be similar to those of IR treatment at steady state. After steady state was achieved with ER galantamine, values for peak concentration and trough concentration were slightly lower (5% and 18%, respectively) than those seen at steady state for IR galantamine; this finding is considered to have no clinical implications. Area under the curve (AUC) with ER galantamine was similar to that seen at steady state with IR galantamine. CONCLUSIONS: These results suggest that no titration period is required in patients receiving stable doses of twice-daily IR galantamine who are switched to once-daily ER galantamine. The once-daily dosage regimen of ER galantamine without a titration period should prove convenient for AD patients and their caregivers and should increase treatment compliance. PMID- 15899096 TI - Association between corneal thickness, mean intraocular pressure, disease stability and severity, and cost of treatment in glaucoma: a Canadian analysis. AB - PURPOSE: We determined the association between the mean corneal thickness (CT) and visual field mean defect (VF) severity as well as with mean intraocular pressure (IOP), disease stability, and cost of glaucoma therapy in a Canadian setting. METHODS: Data were collected from charts of patients diagnosed with primary open-angle glaucoma (POAG). CT measures, VF scores, IOP measurements, physicians' impressions, and resources used (physician visits, diagnostic tests, procedures, and medications) were recorded over a minimum of 2.5 years. CT was compared across the three VF severity levels [mild (0 to < 5 dB), moderate (5 to < 12 dB), and severe (>/= 12 dB)] using a Kruskall-Wallis test. Initial VF was regressed on Age, CT, IOP, and Optic Disc Ratio. Stability and Cost were regressed on IOP. RESULTS: Of the 411 charts, 132 included CT measures. Patients included 50 with mild, 43 with moderate, and 39 with severe disease. The mean CTs of the overall, mild, moderate, and severe groups were 545.9 mum, 554.7 mum, 549.8 mum, and 523.3 mum, respectively. There were statistically significant differences (p < 0.05) between the CT pp of the mild and severe groups as well as between the moderate and severe groups. Regression analyses suggested that CT may be a predictor of disease severity, but not of cost. It was also found that IOP may be a predictor of disease progression. CONCLUSIONS: Patients with severe VFs tend to be those who have thinner corneas. Further research is warranted, as a result of the limited sample size, to clarify the definitive association among corneal thickness, disease progression, and the cost of therapy. PMID- 15899097 TI - Burden of illness in Italian patients with gastro-oesophageal reflux disease. AB - OBJECTIVE: Gastro-oesophageal reflux disease (GORD), characterized by frequent episodes of heartburn, imposes considerable burdens on both patients and health services. In addition to both patients and health services. In addition to gastrointestinal symptoms, GORD may precipitate a variety of extra-oesophageal complications, such as asthma and chest pain, which can significantly impair patients' quality of life. This paper aims to describe the impact of heartburn on patients' health-related quality of life (HRQL) in Italy. RESEARCH DESIGN AND METHODS: Consecutive patients from general practices and gastroenterology clinics who were experiencing heartburn were invited to complete a selection of standardized patient-reported outcomes instruments, including Italian translations of the Gastrointestinal Symptom Rating Scale (GSRS), Short-Form 36 (SF-36), Quality of Life in Reflux and Dyspepsia questionnaire (QOLRAD) and Hospital Anxiety and Depression (HAD) scale. Frequency and severity of heartburn in the preceding week were recorded. RESULTS: Assessments were completed by 152 patients (mean age, 47 years), of whom 60% were female. Sixty-one per cent had moderate symptoms and 74% had symptoms on three or more days in the previous week. Patients were most bothered by reflux, abdominal pain and indigestion. As a result of their symptoms, patients experienced impaired vitality, sleep disturbance and other alimentary problems. This led to impaired HRQL across all but one SF-36 domain. According to the HAD scale, 21% of patients were depressed and 19% were anxious. CONCLUSION: GORD substantially impairs many aspects of HRQL, including sleep, vitality and emotional health. PMID- 15899098 TI - Intraocular pressure lowering effect of brinzolamide 1.0% as adjunctive therapy to latanoprost 0.005% in patients with open angle glaucoma or ocular hypertension: an uncontrolled, open-label study. AB - PURPOSE: A prospective study was conducted to evaluate the intraocular pressure (IOP) lowering effect of brinzolamide 1.0% ophthalmic suspension as an adjunctive therapy with latanoprost 0.005% ophthalmic solution in patients with open angle glaucoma or ocular hypertension. PATIENTS AND METHODS: Fourteen patients with open angle glaucoma (OAG) or ocular hypertension (OH) who had been using latanoprost 0.005% for more than 6 months were initiated on adjunctive brinzolamide therapy. The IOP values at 1 month, 2 months, and 3 months were compared with those measured immediately before adding brinzolamide to the regimen (baseline). The incidence of adverse events such as conjunctival hyperemia and corneal epithelial defect were also examined. RESULTS: The baseline IOP was 21.1 +/- 4.8 mmHg (mean +/- standard deviation). After 1 month, 2 months, and 3 months of therapy IOP was 16.9 +/- 4.5 mmHg, 16.6 +/- 4.0 mmHg, and 15.9 +/ 3.1 mmHg, respectively, showing significant reductions in IOP at all the measuring time-points during the study compared with the baseline value (p < 0.01). Conjunctival hyperemia developed in one patient after 1 month and in another after 2 months; however, both were mild, and therapy was continued. Corneal epithelium defect was observed in 3 patients. One of them had mild defect before brinzolamide was added to the regimen. Increase of eye discharge was seen in one patient. No serious side effects were otherwise observed. CONCLUSION: The addition of brinzolamide to a latanoprost 0.005% regimen may further lower intraocular pressure in patients with open angle glaucoma or ocular hypertension. PMID- 15899099 TI - Management of patients with interstitial cystitis or chronic pelvic pain of bladder origin: a consensus report. AB - BACKGROUND: Interstitial cystitis (IC) remains a challenging disease to treat, but increased awareness, better diagnostic tools, and effective pharmacologic agents can help non-urologists successfully manage this condition. Interstitial cystitis is much more prevalent than previously thought and is often misdiagnosed. Left untreated, IC can have a deleterious effect on the quality of life of the affected individual. In-office symptom-based techniques have been developed to screen for the presence of IC in both women and men presenting with symptoms of chronic pelvic pain (CPP) or chronic prostatitis (CP), and effective pharmacologic treatments exist that can provide acute and long-term symptom relief. Individual patient response to therapy varies, with some patients taking up to several months to achieve an optimal therapeutic response. To optimize patient response, a comprehensive algorithm for the diagnosis and management of IC should be utilized. Symptoms and progress should be assessed at each follow-up visit and changes in therapy made accordingly. Patients should be encouraged to learn as much as possible about IC and to take charge of the management of symptoms using their physician for guidance in decision making. The importance of this aspect of therapy should be reinforced at each subsequent visit. The entire office staff also has an important role in understanding and acknowledging each patient's pain and frustration as well as in providing support and education about IC. SCOPE: This paper presents recommendations from a multidisciplinary panel for the evaluation, diagnosis, treatment, and follow-up care of patients with IC or CPP of bladder origin. PMID- 15899100 TI - Efficacy and tolerability of lumiracoxib 100 mg once daily in knee osteoarthritis: a 13-week, randomized, double-blind study vs. placebo and celecoxib. AB - OBJECTIVE: To determine the efficacy and safety of lumiracoxib for knee osteoarthritis (OA). METHODS: This was a 13-week, multicentre, randomized, double blind, double-dummy, placebo-controlled study. Males or females aged >/= 18 years with primary knee OA received lumiracoxib 100 mg od, lumiracoxib 100 mg od with a loading dose of 200 mg od for the first two weeks, celecoxib 200 mg od, or placebo. MAIN OUTCOME MEASURES: Co-primary variables, assessed at week 13, were OA pain intensity in the target knee, patient's global assessment of disease activity and the WOMAC total score. Other variables included OMERACT-OARSI responder rates and WOMAC subscale scores. Safety and tolerability were evaluated. RESULTS: All active treatments were superior to placebo for all co primary variables. No significant differences were observed between any active treatments. Mean reductions from baseline to week 13 for lumiracoxib 100mg od, 100mg od with loading dose, celecoxib and placebo, respectively, were: OA pain intensity in the target knee: 26.8, 26.2, 26.6 and 21.4mm (all p < 0.01 vs. placebo); patient's global assessment of disease activity: 25.1, 21.9, 22.9 and 18.9 mm (all p < 0.05 vs. placebo); WOMAC total score: 15.2, 14.8, 14.7 and 11.3 (all p < 0.01 vs. placebo). Lumiracoxib was superior to placebo and similar to celecoxib for OMERACT-OARSI response and WOMAC subscale scores. Lumiracoxib was well tolerated. The incidence of adverse events was similar across groups. CONCLUSIONS: Lumiracoxib 100 mg od provided effective relief from the pain of knee OA, with efficacy similar to celecoxib 200 mg od, and was well tolerated. PMID- 15899101 TI - Elevated serum tumor necrosis factor alpha and ferritin may contribute to the insulin resistance found in HCV positive Egyptian patients. AB - OBJECTIVE: There is evidence of an increased incidence of insulin resistance and diabetes mellitus (DM) in patients with hepatitis C virus (HCV) infection. Several mechanisms have been proposed, including inadequate insulin secretion or interference with signaling within the insulin receptor. We assessed serum tumor necrosis factor alpha (TNFalpha) and ferritin levels as potential mediators of insulin resistance in HCV positive Egyptian patients. PATIENTS AND RESULTS: Patients (n = 27) with HCV infection, patients (n = 23) with hepatitis C and DM (HCV + DM), patients (n = 22) with DM, and sex- and age-matched controls (n = 18) were included in this study. The degree of insulin resistance (HOMA index) was significantly higher in the HCV, HCV + DM and DM groups compared to the controls. The mean +/- SD of the HOMA index was 4.53 +/- 2.84, 6.1 +/- 2.36, 3.69 +/- 2.2 and 1.32 +/- 0.49, in HCV, HCV + DM, DM and controls, respectively. Serum TNFalpha levels were significantly higher in the HCV, HCV + DM groups compared with the healthy controls and DM patients (p < 0.001). The median (range) values of TNFalpha in HCV, HCV + DM, DM patients and controls subjects were 25.5 (0.43 124.0), 19.8 (0.51-139), 0.85 (0-10.5) and 0.32 (0-5.8) pg/mL, respectively. There was a significant positive correlation between the HCV load and both HOMA index and TNF alpha level. HCV and HCV + DM patients also had significantly higher serum ferritin levels compared with healthy controls and patients with DM. The mean +/- SD of serum ferritin in HCV, HCV + DM, DM patients and controls subjects was 258.1 +/- 116.2, 285.8 +/- 124.3, 86.9 +/- 41.8 and 159.9 +/- 76.9 ng/mL, respectively. CONCLUSION: Patients with HCV infection had a significantly higher level of TNFalpha and ferritin which may explain their insulin resistance. HOMA index and serum TNFalpha levels correlated positively with the HCV load. PMID- 15899102 TI - The economic impact of GERD and PUD: examination of direct and indirect costs using a large integrated employer claims database. AB - OBJECTIVE: The objective of this study was to examine the relationship of work loss associated with gastro- the relationship of work loss associated with gastro the relationship of work loss associated with gastro-esophageal reflux disease (GERD) and peptic ulcer disease (GERD) and peptic ulcer disease (PUD) in a large population of employed individuals in the United States (US) and quantify the individuals in the United States (US) and quantify the economic impact of these diseases to the employer. METHODS: A proprietary database that contained work place absence, disability and workers' compensation data in addition to prescription drug and medical claims was used to answer the objectives. Employees with a medical claim with an ICD-9 code for GERD or PUD were identified from 1 January 1997 to 31 December 2000. A cohort of controls was identified for the same time period using the method of frequency matching on age, gender, industry type, occupational status, and employment status. Work absence rates and health care costs were compared between the groups after adjusting for demo graphic, and employment differences using analysis of covariance models. RESULTS: There were significantly lower (p < 0.05) prescription, and outpatient costs in the controls compared to the disease groups, although the eta-square values were very low. The mean work absence attributed to sick days was 2.8 (+/- 2.3) for controls, 3.4 (+/ 2.5) for GERD, 3.2 (+/- 2.6) for PUD, and 3.2 (+/- 2.3) days for GERD + PUD. For work loss, a significantly higher (p < 0.05) rate of adjusted all-cause absenteeism and sickness-related absenteeism were observed between the disease groups versus the controls. In particular, controls had an average of 1.2 to 1.6 days and 0.4 to 0.6 lower all-cause and sickness-related absenteeism compared to the disease groups. The incremental economic impact projected to a hypothetical employed population was estimated to be $3441 for GERD, $1374 for PUD, and $4803 for GERD + PUD per employee per year compared to employees without these diseases. CONCLUSIONS: Direct medical cost and work absence in employees with GERD, PUD and GERD + PUD represent a significant burden to employees and employers. PMID- 15899103 TI - How useful are measures of BMD and bone turnover? AB - Measurements of bone mineral density (BMD) and biochemical markers of bone turnover are useful in the diagnosis and management of osteoporosis, as well as in research relating to the pathogenesis and treatment of the disease. Recent challenges to the utility of these measures have resulted in some confusion among both researchers and clinicians. BMD accounts for the great majority of bone strength and is the current gold standard for the diagnosis of osteoporosis, as well as for prediction of fracture risk. Although bone turnover increases sharply after menopause, biochemical markers of bone turnover have limited usefulness in fracture risk prediction. Persistently elevated bone turnover throughout the menopause is associated with structural decrements, cannot be measured routinely and non-invasively. In research applications, both BMD and markers of bone turnover are used to identify candidate agents in preclinical and clinical studies. In addition, head-to-head comparisons of treatments utilize these measures, because fracture endpoint trials would need to be extraordinarily large and complex. Analyses that have suggested that change in BMD or bone turnover 'explains' little of change in fracture risk with treatment appear to be flawed. Although neither can perfectly predict fracture, they are our current best alternatives. PMID- 15899104 TI - Service utilization and costs of olanzapine versus divalproex treatment for acute mania: results from a randomized, 47-week clinical trial. AB - OBJECTIVE: This study examined direct treatment costs based on medication and service use data collected in a 47-week multi-center, double-blind, randomized clinical trial of olanzapine versus divalproex for patients with bipolar disorder and and experiencing acute mania. RESEARCH DESIGN AND METHODS: Patients who completed the 3-week acute phase and entered into the 44-week maintenance phase (n = 147) of the trial were included. Service use data were collected at weeks 3, 7, 15, 23, 31, 39 and 47 of the maintenance phase. Analyses were conducted to address potential biases from discontinuation patterns and use of this patient sub-sample. RESULTS: Overall, per patient yearly costs were similar for olanzapine- and divalproex-treated patients ($14 967 vs. $15 801). Psychiatric related costs accounted for 95.4% and 93.6% of the total costs for olanzapine- and divalproextreated patients, respectively. Study medication costs were significantly higher for olanzapine than for divalproex ($4662 vs. $1755, p < 0.01). However, this was offset by the combined effects of numerically lower costs for several other services with olanzapine treatment. Some of the savings associated with olanzapine treatment compared with divalproex treatment resulted from differences in costs associated with emergency room services ($432 vs. $1346, p < 0.05). CONCLUSIONS: Overall per-patient treatment costs were similar for olanzapine and divalproex. Recognizing challenges in analyzing and generalizing cost outcomes from a clinical trial setting, results provide some much-needed comparative economic information regarding these two medication options for treating mania in bipolar disorder. PMID- 15899105 TI - Post-marketing surveillance: a UK/European perspective. AB - The granting of regulatory approval allows medical practitioners to prescribe a drug in a controlled way to a group of patients defined within the licence. Prior to this, the new product may have been evaluated often in less than 5000 patients and usually in a selected environment in which many patients have been excluded, including for example, women of childbearing potential, the elderly and children. Co-existent disease and the concomitant use of a number of common drug treatments also frequently exclude patients from pre-licensing trials. It is hardly surprising, therefore, that many adverse drug reactions are only detected once the product has been prescribed to the general population. National and international regulatory bodies, therefore, provide systems for post-marketing pharmacosurveillance, although participation in these by clinicians is generally voluntary and under-reporting is widespread. Post-marketing surveillance (PMS) studies are not generally an integral component to launching a new drug and many clinicians are sceptical over data generated in trials which do not conform to the 'gold standard' randomised control trial (RCT) design. However, in dismissing such studies, a great opportunity to obtain information, often from many thousands of subjects, is being missed. This article discusses post-marketing pharmacovigilance and the role of PMS studies in the context of current UK and European legislation. PMID- 15899106 TI - LDL-C goal attainment with the addition of ezetimibe to ongoing simvastatin treatment in coronary heart disease patients with hypercholesterolemia. AB - OBJECTIVE: To evaluate the addition of ezetimibe or placebo to on-going simvastatin treatment on attaining the LDL-C treatment target of 2.60 mmol/L and 100 mg/dL and 18 years) with MDD [baseline scores >/= 18 and 0.05). The ADC values of the test group declined from the fifth week, and after the tenth week the ADC values were significantly different between the test and control groups at gradient factor (b) value 300 sec/mm(2) (P < 0.05). At b value 600 and 1000 sec/mm(2), significant difference was seen between the two groups from the sixth week onward. The range of ADC value of the groups was (1.7 - 0.9) +/- (0.40 - 0.04) mm(2)/sec (b = 600) and (1.38 - 0.75) +/ (0.07 - 0.35) mm(2)/sec (b = 1000), respectively. Dominant pathological changes included swelled hepatocytes within 1 to 4 weeks after the administration of diethylnitrosamine in the test group, hyperplasia of fibrous tissues in 5 - 8 weeks and formation of cirrhotic nodules in 9 - 12 weeks. CONCLUSIONS: MR functional DWI could detect diffuse liver lesions earlier than conventional morphological imaging. ADC value as a marker for early diagnosis of diffuse liver lesions could also be used to inspect changes of the lesions. PMID- 15899119 TI - Modification of the right subclavian vein catheterization and its anatomic basis and techniques. AB - BACKGROUND: Several million subclavian-vein catheters are placed in patients each year to enable caregivers to administer chemotherapy, total parenteral nutrition, or long-term antibiotics or to manage preoperative fluids. Subclavian venipuncture requires the position of a deep vein to be identified with only surface landmarks. But the traditional right subclavian vein (RSV) catheterization (primitive procedures) is not the answer for all patients. The precise location of the vein is not known, and it is important to select the most appropriate method to achieve central venous access safely in any given patient. To modify the primitive procedures of the RSV catheterization for greater success and reduce the complications, anatomic studies and ultrasonography were conducted and clinical applications were validated. METHODS: Anatomical observation and measurement of the RSV and its adjacent structures were performed on 20 adult cadavers according to modified procedures. The RSV catheterization of 2900 cases was carried out by the modified procedure, 500 of these cases were observed by ultrasonography after the operation. RESULTS: The anatomical studies and clinical application showed that the insertion point differs from the bodily form of fatness or leptosome. The clinical data revealed that in the 2900 cases which were performed with the modified approach, the success rate was 98.90% (2868 cases), the failure rate was 1.10% (32 cases), and the complication rate is 0.79% (23 cases), and the catheterization time is (31.2 +/- 10.5) minutes. Five hundred and sixty cases of the RSV catheterization were carried out by the recommended insertion procedure; the results were compared with the modified approach and the traditional approach. The successful rate of the traditional approach was 73.0%, of which the complication rate was 6.1%; the two approaches were significantly different (successful rate: chi(2) = 626.642, P < 0.01; complication rate: chi(2) = 80.708, P< 0.01). CONCLUSIONS: The modified RSV catheterization is characterized with a higher success rate and less complications, and the insertion procedure differs from the bodily form of fatness or leptosome. PMID- 15899120 TI - Liver transplantation for patients with hepatocellular carcinoma at the Liver Cancer Institute of Fudan University, China. AB - BACKGROUND: Selection of patients with hepatocellular carcinoma (HCC) for orthotopic liver transplantation (OLT) remains controversial. Since there is a trend to expand the transplant criteria for HCC patients, we reviewed the data of patients with HCC who had received OLT at our institute to determine their survival and prognostic factors. METHODS: A total of 67 patients with HCC who had undergone OLT from April 2001 through December 2003 were reviewed retrospectively. Selection OLT candidates with HCC was dependent on the anatomical characteristics and/or the severity of underlying liver cirrhosis. The 67 patients were followed up for more than 6 months after transplantation. Their survival rate was calculated by the Kaplan-Meier method. Univariate and multivariate analyses using the Cox proportional hazards regression model were performed to reveal the factors affecting the survival rate. RESULTS: No perioperative death occurred in this series. The 1- and 2-year cumulative survival rates were 90.0% and 65.6%, and the disease-free survival (DFS) rates were 77.5% and 62.5% respectively. Univariate analysis revealed the tumor size, portal vein tumor thrombus (PVTT), serum alpha-fetoprotein level, bilobular distribution of tumors, pTNM stage and histological differentiation were statistically significant factors affecting the DFS (P < 0.05). Multivariate analysis showed tumor size and PVTT were independent and statistically significant factors affecting the DFS (P = 0.005 and 0.010, respectively). In this series, all but 2 received systemic chemotherapy, among them 13 had tumor recurrence within 8 months after OLT. CONCLUSIONS: OLT is indicated for patients with HCC, even for some patients with end-stage liver disease who may survive longer without tumor recurrence. Adjuvant chemotherapy may decrease the recurrence of HCC after OLT. PMID- 15899121 TI - Specific humoral immune responses in rhesus monkeys vaccinated with the Alzheimer's disease-associated beta-amyloid 1-15 peptide vaccine. AB - BACKGROUND: Alzheimer's disease (AD) is a neurodegenerative disorder characterized by overproduction of beta-amyloid (Abeta), with the subsequent pathologic deposition of Abeta which is important for memory and cognition. Recent studies showed murine models of AD and AD patients inoculated with Abeta(1 42) peptide vaccine had a halted or delayed pathological progression of AD. Unfortunately, the clinical phase IIa trial of Abeta(1-42) peptide vaccine (AN1792) was halted prematurely because of episodes of menigoencephalitis in 18 of the vaccinated patients. The vaccination of BALB/c or Tg2576 transgenic mouse with Abeta(1-15) peptide vaccine is safe and the immune effects are satisfactory. This study further characterizes the specific humoral immune responses in adult rhesus monkeys induced by Abeta(1-15) peptide vaccine. METHODS: Five male adult rhesus monkeys were injected intramuscularly with Abeta(1-15) peptide vaccine at baseline and at weeks 2, 6, 10, 14, 18 and 22. The titers and IgG isotypes of the antibody against Abeta(1-42) in serum was measured by Enzyme-linked Immunosorbent Assay (ELISA). The specificity of the antibody against Abeta(1-42) was determined by Western blot. The Abeta plaques in Tg2576 transgenic mouse brain were stained with the antiserum using immunohistochemistry method. RESULTS: At the eighth week after the vaccination, antibody against Abeta(1-42) began to develop significantly in serum. The titers of the antibody increased following vaccine boosted and reached 1:3840 at the twenty-fourth week, then decreased after the termination of inoculation. The IgG1 was accounted for the highest level in the antiserum pool. The antibody against Abeta(1-42) showed high specificity. The Abeta plaques in Tg2576 transgenic mouse brain were labeled with the antiserum. CONCLUSION: Abeta(1-15) vaccine can induce vigorously specific humoral immune responses in adult rhesus monkey. PMID- 15899122 TI - Apoptosis in human germinal centre B cells by means of CC chemokine receptor 3 expression induced by interleukin-2 and interleukin-4. AB - BACKGROUND: CC chemokine receptor 3 (CCR3), expressed on some inflammatory cells, is a member of the chemokine receptor family. Its ligand is eotaxin/CCL11. In this research, we studied the expression and function of CCR3 induced by interleukin-2 (IL-2) and interleukin-4 (IL-4) on human germinal centre (GC) B cells. METHODS: Cells isolated from human tonsils were stimulated with IL-2 or/and IL-4 followed by bonding with eotaxin/CCL11. Flow cytometry was used to detect expression of CCR3 on GC B cells and apoptosis of GC B cells. Real time quantitative reverse transcription polymerase chain reaction and Northern blot assays were used to analyse the CCR3 mRNA expressed in the GC B cells. Chemotaxis and adhesion assays were used to determine the effect of eotaxin/CCL11 ligand bonded to CCR3 on GC B cells. RESULTS: There was no CCR3 expression on human freshly isolated GC B cells. The combination IL-2 and IL-4 could upregulate CCR3 mRNA and protein expression on GC B cells. Eotaxin could not induce GC B cell chemotaxis and adhesion but triggered apoptosis of GC B cells. CONCLUSION: IL-2 and IL-4 together induced expression of CCR3 on GC B cells, and the receptor acted as a death receptor. PMID- 15899123 TI - Phosphorylated extracellular signal-regulated kinase up-regulated p53 expression in shikonin-induced HeLa cell apoptosis. AB - BACKGROUND: The role of extracellular signal-regulated kinase 1/2 (ERK1/2) in shikonin-induced HeLa cells apoptosis remains vague. This study was to investigate the activation of caspase pathways and the role of ERK1/2 in human cervical cancer cells, HeLa, by shikonin. METHODS: The inhibitory effect of shikonin on the growth of HeLa cells was measured by MTT assay. Fluorescent microscopic analysis of apoptotic cells stained with 4',6'-oliiamiclino-2 phenylindole C (DAPI) and Hoechst 33258 was carried out. Caspase-3 and -8 activities were detected using caspase-3 substrate and caspase-8 substrate as substrates, respectively. The protein levels of ERK, p53 and p-ERK were determined by Western blot analysis. RESULTS: Shikonin inhibited cell growth in a time- and dose-dependent manner. Caspase-3 and caspase-8 were activated in the apoptotic process and caspase inhibitors effectively reversed shikonin-induced apoptosis. Phosphorylation of ERK resulted in up-regulation of p53 expression, which was blocked by mitogen-activated protein kinase (MEK), inhibitor PD 98059. CONCLUSION: Shikonin induces HeLa cell apoptosis through the ERK, p53 and caspase pathways. PMID- 15899124 TI - Effects of transforming growth factor beta 1 on the growth of rhabdomyosarcoma cell line RD. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) can inhibit the growth of most epithelial and endothelial cells. The growth regulative role of TGF-beta on soft tissue sarcoma was seldom reported. Here we examined TGF-beta1 effects on the growth of human rhabdomyosarcoma cell RD and searched the relative molecular mechanism. METHODS: The viability of RD was examined by [(3)H]-thymidine incorporation and [3-(4,5-dimethylthiazol-z-yl)-2,5-diphenyl tetrazolium bromide] (MTT) assay. RD cell cycle was analysed by flow cytometry. The protein and mRNA of cell cycle regulative factors in RD were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The kinase activity of cdk2 or cdk4 was examined by immunoprecipitation and kinase assay. Immunofluorescent staining was used to detect the location of cell cycle regulative factors in RD by laser scanning confocal microscope. RESULTS: TGF beta1 inhibits RD proliferation by G1-arrest in cell cycle progression. TGF-beta1 can prominently up-regulate P27 of RD, then augment P27 to bind cyclinE-cdk2 complexes, which effectively suppress cdk2 kinase activity. P21 increased and c myc decreased in RD due to TGF-beta1. Both P15 and cdk4 have not been involved in the growth inhibitory event. TGF-beta1 treatment induced P27 to congregate around nucleus. P21 pervaded from nucleus to both nucleus and cytoplasm by TGF-beta1 treatment. CONCLUSION: TGF-beta1 inhibits the proliferation of human rhabdomyosarcoma cell line RD and induces RD G1-arrest. This course is accomplished by TGF-beta1 up-regulating P27 to suppress cdk2 kinase activity. The induction of P21 and down-regulation of C-myc might participate in the growth arrest event. PMID- 15899125 TI - Establishment and characterization of cell sublines with high and low metastatic potential derived from human osteosarcoma. PMID- 15899126 TI - Adjunctive anti-tachycardia pacing therapy for patients with acute myocardial infarction during emergency percutaneous coronary intervention. PMID- 15899127 TI - Clinical, pathological and genetic study of a kindred of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes. PMID- 15899128 TI - Simultaneous coronary artery bypass grafting, replacement of the innominate artery and subtotal thyroidectomy in a 61 year-old patient: a case-report. PMID- 15899129 TI - Primary endotracheal non-Hodgkin's lymphoma in a Chinese woman: a case report. PMID- 15899130 TI - Variation analysis of the severe acute respiratory syndrome coronavirus putative non-structural protein 2 gene and construction of three-dimensional model. AB - BACKGROUND: The rapid transmission and high mortality rate made severe acute respiratory syndrome (SARS) a global threat for which no efficacious therapy is available now. Without sufficient knowledge about the SARS coronavirus (SARS CoV), it is impossible to define the candidate for the anti-SARS targets. The putative non-structural protein 2 (nsp2) (3CL(pro), following the nomenclature by Gao et al, also known as nsp5 in Snidjer et al) of SARS-CoV plays an important role in viral transcription and replication, and is an attractive target for anti SARS drug development, so we carried on this study to have an insight into putative polymerase nsp2 of SARS-CoV Guangdong (GD) strain. METHODS: The SARS-CoV strain was isolated from a SARS patient in Guangdong, China, and cultured in Vero E6 cells. The nsp2 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into eukaryotic expression vector pCI-neo (pCI neo/nsp2). Then the recombinant eukaryotic expression vector pCI-neo/nsp2 was transfected into COS-7 cells using lipofectin reagent to express the nsp2 protein. The expressive protein of SARS-CoV nsp2 was analyzed by 7% sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The nucleotide sequence and protein sequence of GD nsp2 were compared with that of other SARS CoV strains by nucleotide-nucleotide basic local alignment search tool (BLASTN) and protein-protein basic local alignment search tool (BLASTP) to investigate its variance trend during the transmission. The secondary structure of GD strain and that of other strains were predicted by Garnier-Osguthorpe-Robson (GOR) Secondary Structure Prediction. Three-dimensional-PSSM Protein Fold Recognition (Threading) Server was employed to construct the three-dimensional model of the nsp2 protein. RESULTS: The putative polymerase nsp2 gene of GD strain was amplified by RT-PCR. The eukaryotic expression vector (pCI-neo/nsp2) was constructed and expressed the protein in COS-7 cells successfully. The result of sequencing and sequence comparison with other SARS-CoV strains showed that nsp2 gene was relatively conservative during the transmission and total five base sites mutated in about 100 strains investigated, three of which in the early and middle phases caused synonymous mutation, and another two base sites variation in the late phase resulted in the amino acid substitutions and secondary structure changes. The three-dimensional structure of the nsp2 protein was successfully constructed. CONCLUSIONS: The results suggest that polymerase nsp2 is relatively stable during the phase of epidemic. The amino acid and secondary structure change may be important for viral infection. The fact that majority of single nucleotide variations (SNVs) are predicted to cause synonymous, as well as the result of low mutation rate of nsp2 gene in the epidemic variations, indicates that the nsp2 is conservative and could be a target for anti-SARS drugs. The three-dimensional structure result indicates that the nsp2 protein of GD strain is high homologous with 3CL(pro) of SARS-CoV urbani strain, 3CL(pro) of transmissible gastroenteritis virus and 3CL(pro) of human coronavirus 229E strain, which further suggests that nsp2 protein of GD strain possesses the activity of 3CL(pro). PMID- 15899131 TI - Potent and specific inhibition of SARS-CoV antigen expression by RNA interference. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) is an infectious disease caused by SARS-CoV. There are no effective antiviral drugs for SARS although the epidemic of SARS was controlled. The aim of this study was to develop an RNAi (RNA interference) approach that specifically targeted the N gene sequence of severe acute respiratory syndrome associated coronavirus (SARS-CoV) by synthesizing short hairpin RNA (shRNA) in vivo, and to assess the inhibitory effect of this shRNA on SARS-CoV N antigen expression. METHODS: The eukaryotic expression plasmid pEGFP-C1-N, containing SARS-CoV N gene, was co-transfected into 293 cells with either the RNAi plasmid pshRNA-N or unrelated control plasmid pshRNA-HBV-C4. At 24, 48 and 72 hours post transfection, the green fluorescence was observed through a fluorescence microscope. The RNA levels of SARS-CoV N were determined by reverse transcription polymerase chain reaction (RT-PCR). The expression of Green Fluorescent Protein (GFP) and protein N were detected using Western blot. RESULTS: The vector, pshRNA-N expressing shRNA which targeted the N gene of SARS-CoV, was successfully constructed. The introduction of RNAi plasmid efficiently and specifically inhibited the synthesis of protein N. RT-PCR showed that RNAs of N gene were clearly reduced when the pEGFP-C1-N was cotransfected with pshRNA-N, whereas the control vector did not exhibit inhibitory effect on N gene transcription. CONCLUSIONS: Our results demonstrate that RNAi mediated silencing of SARS-CoV gene could effectively inhibit expression of SARS-CoV antigen, hence RNAi based strategy should be further explored as a more efficacious antiviral therapy of SARS-CoV infection. PMID- 15899132 TI - Epidemiologic study on human immunodeficiency virus infection among children in a former paid plasma donating community in China. AB - BACKGROUND: Illegal plasma collecting activities in mid 1990s caused a large number of human immunodeficiency virus (HIV) infections in rural areas of central eastern China. Although most of these activities have been stopped, there were few reports on secondary transmission from infected former plasma donors to their spouses and from infected mothers to their children. This study was to determine the extent of HIV infections among young children in a rural community with a large proportion of plasma donors. METHODS: A survey was conducted among children aged under 7 years in a former plasma donating community in September 2000: finger blood was collected for HIV antibody testing. Another survey was repeated among children aged under 8 years and their families in the same community in April 2001: urine samples were collected for HIV testing. HIV positive children and samples of HIV negative children, whose mothers were positive based on 2001 survey, were followed up until September 2002 to investigate HIV seroconversion, disease progression and HIV strain analysis. Questionnaires were administered to collect information on children's delivery, breast feeding, medical history and their parents' commercial blood donation history and HIV status. RESULTS: Among 169 children surveyed in 2000, 10 (5.9%) were HIV positive. Of 224 children, 11 were positive in 2001. The overall prevalence rate in the two surveys was 5.0% (17/337) when counting 56 repeated children only once. Of children born to HIV positive mothers, 28.9% were infected. A seroconversion rate of 2.5 per 100 child years was observed by following up 28 HIV negative children. No statistically significant associations were found between children's HIV infection and their histories of blood transfusion, surgery, immunization injection or medical injections. All infections were HIV-1 subtype B' strain, the average dispersion rate is 7.4%. DNA sequence analysis showed a close relationship between the seroconverted children and their infected mothers. CONCLUSIONS: HIV vertical transmissions in the rural former plasma donating community was significant. Intervention measures should be taken to prevent further transmission. It was estimated that the HIV spread in this community occurred in 1994 or even earlier. Many infected people are developing AIDS now: treatment and care are urgently needed for these sick people. PMID- 15899133 TI - Detection and analysis of anti-latent membrane protein 2A antibodies in the sera of patients with Epstein-Barr virus associated malignancies. AB - BACKGROUND: Epstein-Barr virus (EBV) associated malignancies with a Type II latency gene expression pattern, such as Hodgkin's disease, and nasopharyngeal carcinoma (NPC), frequently express the EBV antigen latent membrane protein 2A (LMP2A). We expected to establish a highly expressing LMP2A yeast cell strain and get the high quality LMP2A protein, which was used for detection, analysis and characterization of its antibodies in various patients' sera of EBV associated malignancies. METHODS: The plasmid pPICZalphaA-LMP2A containing the full length of LMP2A cDNA was constructed and transformed to Pichia pastoris GS115 to express LMP2A protein. After fermentation and purification, the LMP2A protein was used as an antigen to detect anti-LMP2A antibodies (Abs) in the sera of patients with EBV associated malignancies in enzyme linked immunosorbent assay (ELISA) or Western blot. RESULTS: LMP2A was expressed successfully with an expected molecular weight of approximately 54 kD and Abs to LMP2A were strikingly specific to NPC. Two thirds or more sera from NPC patients were positive for anti-LMP2A immunoglobulin G (IgG) Abs. The antibodies were absent from the sera of other EBV-associated diseases except a small fraction of the gastric carcinoma. Comparing anti-viral capsid Ags (VCA) IgA and LMP2A IgA titers in the sera from 76 NPC patients, only 55% were positive for anti-LMP2A IgA Abs while 70% were positive for anti-VCA IgA. However, we found that 3 sera negative for VCA IgA were positive for LMP2A IgA. CONCLUSION: The results suggested the potential significance of LMP2A specific Abs for the diagnosis of EBV-associated malignancies, especially NPC. PMID- 15899134 TI - Effect of angiotensin converting enzyme inhibitor on the calcium transients and calcium handling proteins in ventricular myocytes from rats with heart failure. AB - BACKGROUND: Chronic heart failure (CHF) is associated with calcium transients and calcium handling proteins. Angiotensin converting enzyme (ACE) inhibitor has been demonstrated to have beneficial effect on CHF. Yet studies addressed to the relationship between ACE inhibitor and calcium transients in CHF are rare. The aim of this study was to investigate the influence of ACE inhibitor (perindopril) on the contractility and calcium transients and calcium handling proteins in ventricular myocytes from rats with experimental heart failure. METHODS: Male Wistar rats were randomized to heart failure group treated with perindopril [CHF T, 3 mg.kg(-1).d(-1)], heart failure group without treatment (CHF-C) and sham operated group (PS). Heart failure was induced by abdominal aortic constriction. All groups were further followed up for 12 weeks. Left ventricular myocytes were then isolated. Single cell shortening fraction and [Ca(2+)]i were simultaneously measured by laser scanning confocal microscope under the field stimulation (1.0 Hz). Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were performed to evaluate the changes of mRNA and protein of Na(+)-Ca(2+) exchanger (NCX1), sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2) and phospholamban (PLB). RESULTS: The fraction of cell shortening (FS%) and [Ca(2+)]imax (nmol/L) were significantly reduced in group CHF-C compared with group PS (FS%: 7.51 +/- 1.15 vs 13.21 +/- 1.49; [Ca(2+)]i max: 330.85 +/- 50.05 vs 498.16 +/- 14.07; both P < 0.01), and restored at least partially in CHF-T group. In CHF-C group, the left ventricular mRNA of NCX1 and PLB were significantly upregulated in comparing with PS group (RNCX1/beta-Actin: 0.51 +/- 0.12 vs 0.19 +/- 0.06, P < 0.01; RPLB/beta-Actin: 0.26 +/- 0.12 vs 0.20 +/- 0.08, P < 0.05), while SERCA2 mRNA was downregulated (0.48 +/- 0.10 vs 0.80 +/- 0.11, P < 0.01). The mRNA levels of NCX1 and SERCA2 in CHF-T group were between the CHF-C and PS group, and the differences of the latter two groups were significant (all P < 0.05). In CHF-C and CHF-T groups, the protein expression of NCX1 were 1.141 +/- 0.047 and 1.074 +/- 0.081 times of that in PS group respectively (both P < 0.05), and SERCA2 protein levels were 0.803 +/- 0.100 and 0.893 +/- 0.084 times of that in PS group respectively (both P < 0.05). The protein expression of NCX1 and SERCA2 in the CHF-C and CHF-T groups is significantly different (both P < 0.05). CONCLUSION: ACE inhibitor could improve cardiac function of failing heart through directly enhancing the contractility of single cardiomyocyte, and these effects are probably mediated by its roles in preventing the deleterious changes of calcium transients and calcium handling proteins in CHF. PMID- 15899135 TI - Computer construction and analysis of protein models of the mutant gammaD crystallin gene. AB - BACKGROUND: Gammad-crystallin plays an important role in human cataract formation. Being highly stable, gammaD-crystallin proteins are composed of two domains. In this study we constructed and analyzed protein models of the mutant gammaD-crystallin gene, which caused a special fasciculiform congenital cataract affecting a large Chinese family. METHODS: gammaD-crystallin protein structure was predicted by Swiss-Model software using bovine gammaD-crystallin as a template and Prospect software using human betab2-crystallin as a template. The models were observed with a Swiss-Pdb viewer. RESULTS: The mutant gammaD crystallin structure predicted by the Swiss-Model software showed that proline23 was an exposed surface residue and P23T change made a decreased hydrogen bond distance between threonine23 and asparagine49. The mutant gammaD-crystallin structure predicted by the Prospect software showed that the P23T change exerted a significant effect on the protein's tertiary structure and yielded hydrogen bonds with aspartic acid21, asparagine24, asparagine49 and serine74. CONCLUSION: The mutant gammaD-crystallin gene has a significant effect on the protein's tertiary structure, supporting that alteration of gamma-crystallin plays an important role in human cataract formation. PMID- 15899136 TI - Using quantitative CT to predict postoperative pulmonary function in patients with lung cancer. AB - BACKGROUND: At present, the therapy for patients with lung cancer that achieves a high rate of cure is surgical resection at an early stage of the disease. The aim of this study is to evaluate quantitative computed tomography (QCT) for predicting postoperative pulmonary function in patients with lung cancer. METHODS: The data of thirty-one patients with lung cancer who underwent both pulmonary functional tests and QCT scan before operations were collected. A CT program was used to quantify the volume of whole lung parenchyma with attenuation of -910 HU to -600 HU, which was defined as total functional lung volume (TFLV). Similarly, the volume of lung (lobes or segments) with attenuation of -910 HU to 600 HU was defined as regional functional lung volume (RFLV). Forced vital capacity (FVC), forced expiratory volume in first second (FEV1), FVC% and FEV1% (ratio to reference values of the matched population) were obtained from preoperational pulmonary functional tests. According to the formula: predicted FVC (pre-FVC) = preoperative FVC x [1-(RFLV/TFLV)]; predicted FEV1 (pre-FEV1) = preoperative FEV1 x [1-(RFLV/TFLV)], we obtained values of predicted FVC, predicted FEV1, predicted FVC% (pre-FVC/reference values of the matched population), and predicted FEV1% (pre-FEV1/reference values of the matched population). The paired t test and Pearson correlation test were used to assess significance of differences and correlations between CT predicted values and postoperative measured results of FVC, FEV1, FVC% and FEV1%. RESULTS: QCT predicted values correlated well with postoperative FVC, FEV1, FVC% and FEV1% (r = 0.873, 0.809, 0.849 and 0.801 respectively, all P < 0.01). CONCLUSIONS: QCT is an effective and accurate way to predict postoperative pulmonary function in patients undergoing pulmonary resection, regardless of the patients' preoperative pulmonary functional status. PMID- 15899137 TI - Clinical presentation of inflammatory bowel disease: a hospital based retrospective study of 379 patients in eastern China. AB - BACKGROUND: Numerous studies from Europe and North America have provided a wealth of information regarding the epidemiological and clinical characteristics of inflammatory bowel disease (IBD) in Caucasians. Previous studies in mainland China have been limited by small patient numbers or by lack of detailed information about clinical subgroups of the disease. This study was carried out to assess the demographic and clinical characteristics of IBD in Chinese patients. METHODS: In the Sir Run Run Shaw Hospital between 1994 and 2003, 379 patients were diagnosed as IBD. Demographic and clinical data were collected and analysed. RESULTS: Of 379 patients, 317 had ulcerative colitis (UC) (83.6%, 168 male, 149 female, male-female ratio 1.13:1, age range at diagnosis 14-79 years, mean age 44 years) and 62 had Crohn's disease (CD) (16.4%, 39 male and 23 female, male-female ratio 1.70:1, age range at diagnosis 13-70 years, mean age 33 years). In UC, 11.4% of patients had proctitis, 25.2% had proctosigmoiditis, 18.6% were diseased to the splenic flexure and 44.8% had extensive colitis. Nine patients with UC (2.8%) had arthritis, three patients (0.9%) had iritis or conjunctivitis. Of the 62 CD patients, 16 (25.8%) had diseases restricted to the terminal ileum; 15 (24.2%) had colonic diseases; 20 (32.3%) had ileocolonic disease and 11 (17.7%) had disease involving the upper gastrointestinal tract. CONCLUSIONS: This study shows similar characteristics of IBD to that in the West but there are some differences with respect to severity and extraintestinal manifestations. The ethnic and geographic differences may give important clues to the aetiology of IBD. PMID- 15899138 TI - An imaging study using laminin peptide 99mTc-YIGSR in mice bearing Ehrlich ascites tumour. AB - BACKGROUND: The YIGSR is a pentapeptide, from the laminin-1 of the beta1 chain, which can mediate cell adhesion and bind the 67 kD laminin receptor. The purpose is to evaluate the usefulness of (99m)Tc-YIGSR, a novel tumour radiotracer, in the receptor imaging of Ehrlich ascites tumour. METHODS: Using S-acetly-NH3-MAG3 as chelate, YIGSR, a pentapeptide from laminin, was tagged with (99m)Tc. (99m)Tc YIGSR was detected in the tumour group bearing Ehrlich ascites tumour and blocked group. Tumour, normal, inflammatory and blocked groups were imaged. RESULTS: Through reverse phase Sep-Pak C18 chromatogram, it was revealed that YIGSR could conjugate with S-acetly-NH3-MAG3, and be radiolabelled at room temperature and neutral pH with a radiolabelling yield of 62%, and of 4% without chelate. (99m)Tc YIGSR was rapidly cleared from kidney, then liver. The imaging findings showed tumour tissue accumulated initial radioactivity at fifteen minutes after injection in the tumour group, and the uptake increased to peak at three hours with a tumour/muscle ratio (T/M) of 11.36, then cleared slowly to a T/M of 7.50 at eight hours. The tumour uptake of radiotracer in blocked group was significantly lower with T/M of 4.61 at three hours and 0.89 at eight hours. The T/M was only 3.72 at three hours and 1.29 at eight hours after injection in inflammatory group. Compared with inflammatory group and control obstructive group, the ratio of T/M in tumour group was significantly different (P < 0.001). CONCLUSIONS: Using S-acetly-NH3-MAG3, we radiolabelled YIGSR with (99m)Tc. (99m)Tc-YIGSR possesses many merits of tumour imaging: rapid visualization, high sensitivity and specificity, and satisfactory target/nontarget ratio. Our data suggest (99m)Tc-YIGSR is a promising tumour radiotracer. PMID- 15899139 TI - Medical parasitology in China: a historical perspective. PMID- 15899140 TI - Cloning and expression of the fusion protein of interleukin-2 and ESAT6 in Mycobacterium bovis Bacillus Calmette Guerin strain. PMID- 15899141 TI - Immunization effect of purified bivalent vaccine to haemorrhagic fever with renal syndrome manufactured from primary cultured hamster kidney cells. PMID- 15899142 TI - Epidermal growth factor receptor mutations detected in tumors from Chinese "never smokers" with lung adenocarcinoma. PMID- 15899143 TI - Successful transplantation of double unit umbilical-cord blood from unrelated donors in high risk leukemia with a long follow-up. PMID- 15899144 TI - Experimental study of acute lung injury induced by different tidal volume ventilation in rats. PMID- 15899145 TI - Transferrin receptor and Fc alpha/mu receptor may not be the major IgA1 receptor on human mesangial cells. PMID- 15899146 TI - Free vascularized fibular grafting for treatment of old femoral neck fractures. PMID- 15899147 TI - Aplastic anemia associated with dyskeratosis congenita treated with antilymphocyte globulin and cyclosporine: a case report. PMID- 15899148 TI - Potential use of routine databases in health technology assessment. AB - OBJECTIVES: To develop criteria for classifying databases in relation to their potential use in health technology (HT) assessment and to apply them to a list of databases of relevance in the UK. To explore the extent to which prioritized databases could pick up those HTs being assessed by the National Coordinating Centre for Health Technology Assessment (NCCHTA) and the extent to which these databases have been used in HT assessment. To explore the validation of the databases and their cost. DATA SOURCES: Electronic databases. Key literature sources. Experienced users of routine databases. REVIEW METHODS: A 'first principles' examination of the data necessary for each type of HT assessment was carried out, supplemented by literature searches and a historical review. The principal investigators applied the criteria to the databases. Comments of the 'keepers' of the prioritized databases were incorporated. Details of 161 topics funded by the NHS R&D Health Technology Assessment (HTA) programme were reviewed iteratively by the principal investigators. Uses of databases in HTAs were identified by literature searches, which included the title of each prioritized database as a keyword. Annual reports of databases were examined and 'keepers' queried. The validity of each database was assessed using criteria based on a literature search and involvement by the authors in a national academic network. The costs of databases were established from annual reports, enquiries to 'keepers' of databases and 'guesstimates' based on cost per record. For assessing effectiveness, equity and diffusion, routine databases were classified into three broad groups: (1) group I databases, identifying both HTs and health states, (2) group II databases, identifying the HTs, but not a health state, and (3) group III databases, identifying health states, but not an HT. Group I datasets were disaggregated into clinical registries, clinical administrative databases and population-oriented databases. Group III were disaggregated into adverse event reporting, confidential enquiries, disease-only registers and health surveys. RESULTS: Databases in group I can be used not only to assess effectiveness but also to assess diffusion and equity. Databases in group II can only assess diffusion. Group III has restricted scope for assessing HTs, except for analysis of adverse events. For use in costing, databases need to include unit costs or prices. Some databases included unit cost as well as a specific HT. A list of around 270 databases was identified at the level of UK, England and Wales or England (over 1000 including Scotland, Wales and Northern Ireland). Allocation of these to the above groups identified around 60 databases with some potential for HT assessment, roughly half to group I. Eighteen clinical registers were identified as having the greatest potential although the clinical administrative datasets had potential mainly owing to their inclusion of a wide range of technologies. Only two databases were identified that could directly be used in costing. The review of the potential capture of HTs prioritized by the UK's NHS R&D HTA programme showed that only 10% would be captured in these databases, mainly drugs prescribed in primary care. The review of the use of routine databases in any form of HT assessment indicated that clinical registers were mainly used for national comparative audit. Some databases have only been used in annual reports, usually time trend analysis. A few peer-reviewed papers used a clinical register to assess the effectiveness of a technology. Accessibility is suggested as a barrier to using most databases. Clinical administrative databases (group Ib) have mainly been used to build population needs indices and performance indicators. A review of the validity of used databases showed that although internal consistency checks were common, relatively few had any form of external audit. Some comparative audit databases have data scrutinised by participating units. Issues around coverage and coding have, in general, received little attention. NHS funding of databases has been mainly for 'Central Returns' for management purposes, which excludes those databases with the greatest potential for HT assessment. Funding for databases was various, but some are unfunded, relying on goodwill. The estimated total cost of databases in group I plus selected databases from groups II and III has been estimated at pound 50 million or around 0.1% of annual NHS spend. A few databases with limited potential for HT assessment account for the bulk of spending. CONCLUSIONS: Suggestions for policy include clarification of responsibility for the strategic development of databases, improved resourcing, and issues around coding, confidentiality, ownership and access, maintenance of clinical support, optimal use of information technology, filling gaps and remedying deficiencies. Recommendations for researchers include closer policy links between routine data and R&D, and selective investment in the more promising databases. Recommended research topics include optimal capture and coding of the range of HTs, international comparisons of the role, funding and use of routine data in healthcare systems and use of routine database in trials and in modelling. Independent evaluations are recommended for information strategies (such as those around the National Service Frameworks and various collaborations) and for electronic patient and health records. PMID- 15899149 TI - Clinical and cost-effectiveness of newer immunosuppressive regimens in renal transplantation: a systematic review and modelling study. AB - OBJECTIVES: To examine the clinical effectiveness and cost-effectiveness of the newer immunosuppressive drugs for renal transplantation: basiliximab, daclizumab, tacrolimus, mycophenolate (mofetil and sodium) and sirolimus. DATA SOURCES: Electronic databases. Industry submissions. Current Clinical Trials register. Cochrane Collaboration Renal Disease Group. REVIEW METHODS: The review followed the InterTASC standards. Each of the five company submissions to the National Institute for Clinical Excellence (NICE) contained cost-effectiveness models, which were evaluated by using a critique covering (1) model checking, (2) a detailed model description and (3) model rerunning. RESULTS: For induction therapy, three randomised controlled trials (RCTs) found that daclizumab significantly reduced the incidence of biopsy-confirmed acute rejection and patient survival at 6 months/1 year compared with placebo, but not compared with the monoclonal antibody OKT3. There was no significant gain in patient survival or graft loss at 3 years. The incidence of side-effects with daclizumab reduced compared to OKT3. Eight RCTs found that basiliximab significantly improved 6 month/1-year biopsy-confirmed acute rejection compared to placebo, but not compared to either ATG or OKT3. There was no significant gain in either 1-year patient survival or graft loss. The incidence of side-effects with basiliximab was not significantly different compared to OKT3/ATG. For initial/maintenance therapy, 13 RCTs found that tacrolimus reduced the 6-month/1-year incidence of biopsy-proven acute rejection compared to ciclosporin. There was no significant improvement in either 1-year or long-term (up to 5 years) graft loss or patient survival. The acute rejection benefit of tacrolimus over ciclosporin appeared to be equivalent for Sandimmun and Neoral. There were important differences in the side-effect profile of tacrolimus and ciclosporin. Seven RCTs found that mycophenolate mofetil (MMF) reduced the incidence of acute rejection. There was no significant difference in patient survival or graft loss at 1-year or 3-year follow-up. There appeared to be differences in the side-effect profiles of MMF and azathioprine (AZA). No RCTs comparing MMF with AZA were identified. One RCT compared mycophenolate sodium (MPS) to MMF and reported no difference between the two drugs in 1-year acute rejection rate, graft survival, patient survival or side-effect profile. Two RCTs suggest that addition of sirolimus to a ciclosporin based initial/maintenance therapy reduces 1-year acute rejections in comparison to a ciclosporin (Neoral) dual therapy alone and substituting azathioprine with sirolimus in initial/maintenance therapy reduces the incidence of acute rejection. Graft and patient survival were not significantly different with either sirolimus regimen. Adding sirolimus increases the incidence of side effects. The side-effect profiles of azathioprine and sirolimus appear to be different. For the treatment of acute rejection, three RCTs suggested that both tacrolimus and MMF reduce the incidence of subsequent acute rejection and the need for additional drug therapy. Only one RCT and one subgroup analysis in children (<18 years) were identified comparing ciclosporin to tacrolimus and sirolimus, respectively. CONCLUSIONS: The newer immunosuppressant drugs (basiliximab, daclizumab, tacrolimus and MMF) consistently reduced the incidence of short-term (1-year) acute rejection compared with conventional immunosuppressive therapy. The independent use of basiliximab, daclizumab, tacrolimus and MMF was associated with a similar absolute reduction in 1-year acute rejection rate (approximately 15%). However, the effects of these drugs did not appear to be additive (e.g. benefit of tacrolimus with adjuvant MMF was 5% reduction in acute rejection rate compared with 15% reduction with adjuvant AZA). Thus, the addition of one of these drugs to a baseline immunosuppressant regimen was likely to affect adversely the incremental cost-effectiveness of the addition of another. The trials did not assess how the improvement in short-term outcomes (e.g. acute rejection rate or measures of graft function), together with the side effect profile associated with each drug, translated into changes in patient related quality of life. Moreover, given the relatively short duration of trials, the impact of the newer immunosuppressants on long-term graft loss and patient survival remains uncertain. The absence of both long-term outcome and quality of life from trial data makes assessment of the clinical and cost-effectiveness on the newer immunosuppressants contingent on modelling based on extrapolations from short-term trial outcomes. The choice of the most appropriate short-term outcome (e.g. acute rejection rate or measures of graft function) for such modelling remains a matter of clinical and scientific debate. The decision to use acute rejection in the meta-model in this report was based on the findings of a systematic review of the literature of predictors of long-term graft outcome. Only a very small proportion of the RCTs identified in this review assessed patient-focused outcomes such as quality of life. Since immunosuppressive drugs have both clinical benefits and specific side-effects, the balance of these harms and benefits could best be quantified through future trials using quality of life measures. The design of future trials should be considered with a view to the impact of drugs on particular renal transplant groups, particularly higher risk individuals and children. Finally, there is a need for improved reporting of methodological details of future trials, such as the method of randomisation and allocation concealment. A number of issues exist around registry data, for example the use of multiple drug regimens and the need to assess the long-term outcomes. An option is the use of observational registry data including, if possible, prospective data on all consecutive UK renal transplant patients. Data capture for each patient should include immunosuppressant regimens, clinical and patient-related outcomes and patient demographics. PMID- 15899150 TI - Effects of supplementary food on the winter inhibition of reproduction in male and female four-striped field mice (Rhabdomys pumilio). AB - The effects of winter food supplementation on reproduction in the seasonally breeding four-striped field mouse Rhabdomys pumilio were investigated at Mountain Zebra National Park in the Eastern Cape Province of South Africa. On both control and supplemented grids, reproductive activity in females was inhibited; no pregnant females were collected and juveniles were only present in the first winter month. The provision of additional food resulted in an increase in body mass and mass of the male and female reproductive organs. However, all males, from both grids, were spermatogenically active. Ovarian activity was not stimulated by the provision of additional food, but the development of the uterus was and the endometrium was thicker and more vascularised in mice from the supplemented grid than from the control grid. We conclude that seasonal reproduction in R. pumilio is controlled by the females, in which reproductive activity is inhibited in winter. However, the provision of supplementary food was not sufficient to override the reproductive inhibition. PMID- 15899151 TI - The resin monomer triethylene glycol dimethacrylate exhibits reproductive toxicity in male mice. AB - The present study evaluated the effect of triethyleneglycol dimethacrylate (TEGDMA) on mouse fertility. Adult male mice were administered TEGDMA (0, 25 and 100 microg kg(-1)) intragastrically daily for 28 days and then allowed to mate for 10 days with sexually mature untreated female mice; fertility was assessed after a further 10 days. Females mated by males that had been treated with TEGDMA had a significant reduction in pregnancy rate, as well as a significant increase in the total number of resorptions as a percentage of the total number of implantations. Furthermore, the number of animals in which resorption occured was increased significantly. Males treated with TEGDMA exhibited a significant reduction in bodyweight, testis weight and the weight of the seminal vesicles and preputial glands. Testicular and epididymal sperm counts, as well as the efficiency of sperm production, were also significantly reduced in TEGDMA-treated males. The results suggest that TEGDMA has adverse effects on the fertility and reproductive system of male mice. PMID- 15899152 TI - Effect of hexoses and gonadotrophin supplementation on bovine oocyte nuclear maturation during in vitro maturation in a synthetic follicle fluid medium. AB - In vitro oocyte maturation (IVM) culture conditions have been relatively unchanged over the past few decades and remain suboptimal. In contrast, studies of the in vivo environment have led to significant improvements to in vitro embryo culture technologies. The aim of the present study was to determine the effect of maturing bovine cumulus-oocyte complexes (COCs) in medium based on the composition of bovine follicular fluid (Bovine VitroMat; Cook Australia, Eight Mile Plain, Qld, Australia). In particular, the effect of different glucose concentrations and glucosamine supplementation on meiotic maturation was determined. Culturing COCs in the presence of gonadotrophins in Bovine VitroMat, containing either physiological glucose concentrations (2.3 mM) or 5.6 mM (equivalent to levels in Tissue Culture Medium 199 (TCM199)) supplemented with glucosamine resulted in comparable cumulus expansion to COCs cultured in TCM199 plus gonadotrophins. However, nuclear maturation was 1.3-fold lower in Bovine VitroMat cultures containing 2.3 mM glucose compared with 5.6 mM glucose and this effect was independent of glucosamine supplementation. Investigations into the effects of different glucose concentrations and gonadotrophin supplementation during the initial 6 h of maturation demonstrated that COCs cultured in Bovine VitroMat with 5.6 mM glucose without gonadotrophins had a twofold acceleration of the rate of meiotic resumption, yet the rate of polar body formation was decreased by approximately 20% compared with cultures in 2.3 mM glucose and TCM199. However, this effect was not seen when COCs were cultured for the initial 16 h in Bovine VitroMat + 5.6 mM minus gonadotrophins or in Bovine VitroMat + 2.3 mM glucose +/- gonadotrophins. These data demonstrate that glucose concentrations and the timing of the introduction of gonadotrophin during IVM have variable effects on nuclear maturation. Manipulation of glucose concentrations may be a mechanism to influence oocyte meiotic progression and may lead to the development of improved IVM systems, allowing for an increased developmental capacity of bovine oocytes. PMID- 15899153 TI - Embryo transfer in the dromedary camel (Camelus dromedarius) using asynchronous, meclofenamic acid-treated recipients. AB - A total of 40 (Day 7) embryos were recovered from the uteri of 10 superovulated camels. Recipient camels (n = 30) were prepared by injection with 20 (1/4)g of the gonadotrophin-releasing hormone analogue buserelin (i.v.) to induce ovulation and then treatment with 1 g meclofenamic acid (a prostaglandin synthetase inhibitor), orally, once on Day 7 and twice daily on Days 8 and 9 after ovulation and thereafter at a dose of 1 g day(-1) until 8 days after embryo transfer. Embryos were transferred into recipients on Day 8 (n = 10), Day 10 (n = 10) or Day 12 (n = 10) after ovulation and another 10 embryos were transferred into untreated recipients on Day 8 after ovulation as controls. In addition, serum samples from all recipient camels were recovered daily throughout the period of meclofenamic acid administration and for a further 7 days after treatment had ceased and were assayed for progesterone concentrations. Results showed that whereas only one of 10 of the control group of recipients (10%) was diagnosed pregnant, a total of eight of 10 Day 8 (80%), six of 10 Day 10 (60%) and seven of 10 Day 12 (70%) recipients were diagnosed pregnant by ultrasonography 12 days after the embryo had been transferred. Subsequently, however, four pregnancies were lost when the conceptus was aged between 22 and 60 days, but this is not considered above the early fetal mortality rate expected in camels after natural mating or after transferring camel embryos to untreated recipients. Serum progesterone concentrations remained elevated, above 2 ng mL(-1), throughout the period of meclofenamic acid administration in all recipient camels; thereafter, concentrations remained above 2 ng mL(-1) in pregnant animals, whereas in non pregnant camels concentrations had declined to baseline values (<1 ng mL(-1)) within 3 days of the end of the treatment period. In conclusion, treatment of recipient camels with meclofenamic acid reduced the need for tightly timed synchrony between donor and recipient because pregnancies were established in recipients that had ovulated as much as 5 days ahead of the donor. PMID- 15899154 TI - 15-Deoxy-Delta12,14-prostaglandin J2 and peroxisome proliferator-activated receptor gamma (PPARgamma) levels in term placental tissues from control and diabetic rats: modulatory effects of a PPARgamma agonist on nitridergic and lipid placental metabolism. AB - 15-Deoxy-Delta(12,14)-prostaglandin J2 (15dPGJ2) is a peroxisome proliferator activated receptor (3) (PPAR(3)) ligand that regulates lipid homeostasis and has anti-inflammatory properties in many cell types. We postulated that 15dPGJ2 may regulate lipid homeostasis and nitric oxide (NO) levels in term placental tissues and that alterations in these pathways may be involved in diabetes-induced placental derangements. In the present study, we observed that, in term placental tissues from streptozotocin-induced diabetic rats, 15dPGJ2 concentrations were decreased (83%) and immunostaining for nitrotyrosine, indicating peroxynitrite induced damage, was increased. In the presence of 15dPGJ2, concentrations of nitrates/nitrites (an index of NO production) were diminished (40%) in both control and diabetic rats, an effect that seems to be both dependent on and independent of PPAR(3) activation. Exogenous 15dPGJ2 did not modify lipid mass, but decreased the incorporation of (14)C-acetate into triacylglycerol (35%), cholesteryl ester (55%) and phospholipid (32%) in placenta from control rats, an effect that appears to be dependent on PPAR(3) activation. In contrast, the addition of 15dPGJ2 did not alter de novo lipid synthesis in diabetic rat placenta, which showed decreased levels of PPAR(3). We conclude that 15dPGJ2 modulates placental lipid metabolism and NO production. The concentration and function of 15dPGJ2 and concentrations of PPAR(3) were altered in placentas from diabetic rats, anomalies probably involved in diabetes-induced placental dysfunction. PMID- 15899155 TI - Macrophage migration inhibitory factor suppresses transforming growth factor beta2 secretion in cultured rat testicular peritubular cells. AB - Cytokines have direct effects on testicular cell functions and a number of cytokines are produced constitutively within the testis, even in the absence of immune-activation events. There is clear evidence that cytokines play a dual role as important regulatory factors in the normal function of the testis, as well as in testicular inflammation. The pro-inflammatory cytokine macrophage migration inhibitory factor (MIF) is expressed locally in the testis and has direct effects on peritubular cells, which, in turn, produce anti-inflammatory mediators, including transforming growth factor (TGF)-(2)2. In the present study, we investigated the function of MIF by examining its effect on the secretion of TGF (2)2 in peritubular cells. Expression of TGF-(2)2 mRNA was shown by reverse transcription-polymerase chain reaction in peritubular cells isolated from 19-day old rat testis. The addition of recombinant MIF to cultured peritubular cells resulted in a dose-dependent decrease in TGF-(2)2 secretion up to 52% of control levels after 48 h, which was significant for all doses investigated (10-100 ng mL(-1) MIF). Inhibition of TGF-(2)2 secretion was sustained for 72 h for the highest dose of MIF used (100 ng mL(-1)). No effect of MIF was observed on TGF (2)2 mRNA expression levels, as shown by real-time polymerase chain reaction. These results suggest that the pro-inflammatory cytokine MIF can shift the cytokine balance from the immunosuppressive state towards an inflammatory reaction, potentially through the inhibition of TGF-(2)2 secretion by peritubular cells. PMID- 15899156 TI - Major plasma proteins in pig serum during postnatal development. AB - The time-course of changes in the levels of albumin, alpha-fetoprotein (AFP), alpha(1)-protease inhibitor (alpha(1)-antitrypsin), alpha(1)-acid glycoprotein, fetuin, haptoglobin, transferrin, IgG and the major acute-phase protein (Pig-MAP) in the blood sera of pigs during the first days and weeks of life was investigated by quantitative radial immunodiffusion. The serum of newborn pigs before suckling was characterised by a very low concentration of total proteins (approximately 25 mg mL(-1)), low levels of albumin and transferrin and the lack of immunoglobulins. In contrast, alpha(1)-acid glycoprotein and fetuin are present at high levels (approximately 12 and 5 mg mL(-1) respectively). The results of the present study show that the piglets undergo a very rapid metabolic maturation with regard to serum proteins, evolving from a characteristic 'fetal' pattern to an 'adult' one. We have paid special attention to the evolution of haptoglobin and Pig-MAP, which are two important acute-phase proteins in pigs. The evolution of serum levels of these proteins suggests that piglets must overcome a moderate acute-phase situation during the first week of life. PMID- 15899157 TI - Stage-dependent expression of extra-embryonic tissue-spermatogenesis-homeobox gene 1 (ESX1) protein, a candidate marker for X chromosome-bearing sperm. AB - Extra-embryonic tissue-spermatogenesis-homeobox gene 1 (Esx1) encodes an X-linked homeobox protein. Despite the fact that the temporal and spatial mRNA expression pattern of the protein has been studied extensively in the testis, specific localisation of ESX1 in the testis remains to be determined. In the present study, we generated ESX1 antiserum to investigate the stage- and tissue-specific expression of ESX1 in the mouse. Western blotting and immunofluorescent analyses revealed that general localisations of ESX1 were consistent with its RNA expression patterns; that is, it was restricted mainly to the placenta and testis. Immunofluorescent studies demonstrated that ESX1 existed in the testes after 3 weeks of age, coincident with the appearance of round spermatids in the seminiferous tubules. Moreover, ESX1 expression became more abundant in the luminal regions of the seminiferous tubules as the development of round spermatids progressed into spermatozoa. In contrast, reduced expression of ESX1 was observed in experimentally induced cryptorchid testes. The later expression of ESX1 suggests a role in post-meiotic germ cell development. To further understand ESX1 expression in sperm with respect to X chromosome-bearing sperm, we used ESX1 antiserum to immunostain sperm by confocal laser microscopy. Approximately half the sperm population was recognised by the ESX1 antiserum. On the basis of results of the present study, we suggest that ESX1 could be used as a protein marker for X chromosome-bearing sperm. PMID- 15899158 TI - Follicular expression of a human beta-cell leukaemia/lymphoma-2 (Bcl-2) transgene does not decrease atresia or increase ovulation rate in swine. AB - Transgenic (TG) gilts carrying a human Bcl-2 cDNA transgene driven by mouse inhibin-alpha subunit promoter were produced and evaluated to determine if ectopic expression of Bcl-2 in the ovaries would decrease the frequency of atresia in antral follicles and increase ovulation rate. Immunohistochemical analysis showed that the Bcl-2 transgene protein was expressed in granulosa and theca cells, in 86% of healthy and 54% of atretic follicles analysed in TG prepubertal and Day 50 pregnant gilts combined (n = 24). In contrast, Bcl-2 transgene protein was expressed in only 1.4% of healthy and 0% of atretic follicles in non-TG littermates (n = 13). Real-time reverse transcription polymerase chain reaction analysis confirmed that human Bcl-2 was expressed in follicles of TG gilts. The atresia rate for the TG and non-TG groups did not differ (P > 0.05) for prepubertal (45 v. 59%) and Day 50 pregnant gilts (53 v. 52%) respectively. The mean +/- s.e.m. ovulation rate did not differ (P > 0.5) between TG (15.9 +/- 0.8, n = 12) and non-TG (16.4 +/- 0.6, n = 7) Day 50 pregnant gilts. The molecular basis of the failure of ectopic Bcl-2 expression to increase the ratio of healthy to atretic follicles is unknown, but it is possible that the activity of the mitochondrial-dependent cell death pathway was not neutralized by ectopic expression of human Bcl-2 or that other cell death pathways compensated for the decreased mitochondrial-dependent cell death. PMID- 15899159 TI - Effects of hypothermic liquid storage and cryopreservation on basal and induced plasma membrane phospholipid disorder and acrosome exocytosis in boar spermatozoa. AB - Flow cytometry was utilised to determine whether short-term (Day 1) or long-term hypothermic liquid storage (Day 5), or cryopreservation of boar spermatozoa (1) caused changes in plasma membrane phospholipid disorder (MPLD) and acrosome exocytosis (AE), indicative of an advanced stage of capacitation or acrosome status, and (2) facilitated or inhibited the induction of capacitation and the acrosome reaction. Merocyanine with Yo-Pro-1 and peanut agglutinin-fluorescein isothiocyanate with propidium iodide were used to identify MPLD and AE, respectively, in viable spermatozoa. The incidence of basal sperm MPLD and AE in fresh semen was very low (1.1 and 2.2%, respectively) and was increased (P < 0.05) only a small amount in Day 5 and cryopreserved semen (3-8%). Compared to no bicarbonate, incubation with bicarbonate increased MPLD, but the response was greatest (P < 0.05) in fresh sperm (52.3%) compared with Day 1 (36.6%), Day 5 (13.9%) and cryopreserved sperm (13.6%). Incubation with calcium ionophore A23187 increased AE in spermatozoa, but the response was less (P < 0.05) for fresh (34%) and cryopreserved (27%) semen than for Day 1 (45%) and Day 5 (57%) semen. In summary, hypothermic liquid storage and cryopreservation of boar spermatozoa did not advance capacitation or acrosome status in viable spermatozoa, but did alter their responses to induction of capacitation and the acrosome reaction. PMID- 15899160 TI - Fertility aspects in yearling beef bulls grazing endophyte-infected tall fescue pastures. AB - During a 2-year study, yearling beef bulls were used to determine the effects of grazing on endophyte-infected tall fescue on endocrine profiles, semen quality and fertilisation potential. Bulls were allotted to graze tall fescue pastures infected with Neotyphodium coenophialum (E+; n = 20 per year) or Jesup/MaxQ (Pennington Seed, Atlanta, GA, USA; NTE; n = 10 per year). Bulls were grouped by scrotal circumference (SC), bodyweight (BW), breed composites and age to graze tall fescue pastures from mid-November until the end of June (within each year). Blood samples, BW, SC and rectal temperatures (RT) were collected every 14 days. Semen was collected from bulls every 60 days by electroejaculation and evaluated for motility and morphology. The developmental competence of oocytes fertilised in vitro with semen from respective treatments was determined. Bulls grazing E+ pastures had decreased BW gain (P < 0.01), increased overall RT (P < 0.01) and decreased prolactin (P < 0.01) compared with animals grazing NTE pastures. Neither percentage of normal sperm morphology nor motility differed between bulls grazed on the two pasture types. Semen from E+ bulls demonstrated decreased cleavage rates (P = 0.02) compared with semen from NTE bulls. However, development of cleaved embryos to the eight-cell and blastocyst stages did not differ between the two groups. In conclusion, semen from bulls grazing E+ tall fescue resulted in decreased cleavage rates in vitro, which may lower reproductive performance owing to reduced fertilisation ability. PMID- 15899161 TI - [Previous infection with Chlamydia pneumoniae and long-term prognosis in patients with acute coronary syndrome with non-ST segment elevation]. AB - BACKGROUND AND OBJECTIVE: Contradictory data exists from case-control studies and in patients with stable coronary artery disease on the association of prior exposure to Chlamydia pneumoniae and cardiovascular events. We underwent a prospective study to investigate the prognostic value of C. pneumoniae seropositivity in patients with acute coronary syndromes. PATIENTS AND METHOD: In a prospective cohort of 259 consecutive patients (194 men and 65 women), mean age 65 (10 years) with non-ST elevation acute coronary syndromes, we measured serum levels of IgG antibodies directed against C. pneumoniae. RESULTS: After a mean follow-up of 28 (25, 29) months, the incidence of cardiovascular death or myocardial infarction was of 15% in seropositive patients versus 13% in seronegatives at IgG titers (1:64 (p=0.58); of 14% versus 14% at IgG titers > or = 1:128 (p=0.96); and of 14% versus 15% at IgG titers (1:256 (p=0.82). The relative risks (RR, 95% CI) of these major cardiac events adjusted for possible confounding factors were 1.11 (0.52-2.40); 1.01 (0.52-1.96); and 0.94 (0.48-1.87) respectively. CONCLUSIONS: Chlamydia pneumoniae IgG seropositivity is not associated with a higher incidence of death or myocardial infarction in patients with non-ST segment elevation acute coronary syndromes. PMID- 15899162 TI - Exercise capacity in sarcoidosis. Study of 29 patients. AB - BACKGROUND AND OBJECTIVE: Sarcoidosis is a systemic granulomatous disease of unknown etiology. Aims of this prospective study are to evaluate degree of impairment in pulmonary function tests (PFT), arterial blood gas analysis (ABG), respiratory muscle strength, exercise capacity and correlation of these parameters with radiological stages; to further evaluate the use of cardiopulmonary exercise testing in assessment of extent of pulmonary disease; and to discuss the pathophysiologic mechanisms of limitation in exercise capacity in patients with sarcoidosis. PATIENTS AND METHOD: 29 patients with sarcoidosis were grouped according to their radiological stages (stage I: group 1; stage II, group 2; stage III, group 3). Groups 1, 2 and 3 included 11, 13 and 5 patients, respectively. PFT, cardiopulmonary exercise testing and ABG were performed for each patient. RESULTS: Evaluation of all patients showed a significant decrement in exercise capacity. Patients in stage III had decreased diffusing capacity and exercise capacity. There was limitation in exercise capacity in stage I patients who had completely normal spirometry and diffusing capacity. We also found a correlation between radiological stages of the disease and exercise capacity, diffusing capacity and ABG. CONCLUSIONS: Exercise capacity is impaired also in early stages of sarcoidosis and it was found to be the earliest impaired physiological parameter in sarcoid patients. Exercise intolerance, having mutifactorial basis, is correlated with radiological stages. Circulatory impairment and impaired heart rate response to exercise have effects on limitation in exercise capacity. Especially in advanced radiological stages of disease, ventilatory and gas exchange impairment also seems to be effective on limiting exercise in patients with sarcoidosis. PMID- 15899163 TI - [Oral iron and folic acid supplements in a preoperative autologous blood collection program: a randomized study]. AB - BACKGROUND AND OBJECTIVE: We evaluated the capacity of oral iron with or without oral folic acid administration to improve the accomplishment of our scheduled preoperative autologous blood collection program in patients with baseline hemoglobin > 115 g/l. PATIENTS AND METHOD: Patients were enrolled in a randomized trial. The control group received no vitamin supplements. The iron group received 105 mg of elemental iron daily p.o. The and iron+folate group received 105 mg of elemental iron daily and 5 mg of folic acid daily p.o. RESULTS: Eighty-six percent of patients in the control group, 86% of patients in the iron group and 87% of patients in the iron+folate group accomplished our preoperative autologous blood collection program. CONCLUSION: In our study, neither oral iron nor folic acid supplements enhanced the accomplishment of our preoperative autologous blood collection program in patients with baseline hemoglobin > 115 g/l. PMID- 15899164 TI - [Relation between intermediate troponin T values and diagnosis of cardiac disease]. AB - BACKGROUND AND OBJECTIVE: Troponins are useful tools in management of patients with acute coronary syndrome (ACS). Small increases do not guarantee a desirable precision (CV < 10%) and there is a "grey zone" between 99 percentile and clinical cut off. Our objective was to evaluate if "grey zone" troponin values reflect heart damage or they are only "analyzer noise". PATIENTS AND METHOD: Final diagnoses of patients with "grey zone" values attended during 2001 and 2002 were reviewed (group Tn). Control group (Cn): patients with negative values (four months randomly chosen during the same period). Admitted patients were classified in: A, ACS; B, non ischemic heart disease, and C, non heart disease. RESULTS: Tn group (n = 521): admitted, 330 (63.3%); A (53.9%), B (30.9%), C (15.2%). Cn group (n = 652): admitted, 156 (23.9%); A (41%), B (24.4%), C (34.6%). Odds ratio for admittance (Tn vs Cn) was 4.82 (95% CI, 3.68-6.31); OR for diagnosis A, B and C (Tn vs Cn) were: A (1.56) (95% CI, 1.04-2.34), B (1.43) (95% CI, 0.90-2.25), C (0.38) (95% CI, 0.24-0.6). CONCLUSIONS: Intermediate troponin values are associated with diagnosis of heart disease, especially ACS. PMID- 15899165 TI - [Usefulness of pharmacokinetics in the management of antiretroviral therapy]. PMID- 15899166 TI - [Indications and consequences of Helicobacter pylori eradication on gastroesophageal reflux disease]. AB - Several epidemiological data indicate that H. pylori infection prevalence in patients with gastroesophageal reflux disease (GERD) is lower than that reported in respective controls, which would suggest that the organism plays a protective role against this disease. On the other hand, most studies demonstrate that the presence of the infection in patients with GERD does not negatively affect the therapeutic efficacy of proton pump inhibitors (PPIs), and, in case of negatively influencing it, the effects are not clinically relevant and are easily controllable with standard antisecretory treatment. Therefore, the decision to administer H. pylori eradication treatment to a patient should not be influenced by the concomitant presence of GERD. In most cases, H. pylori eradication does not seem to induce GERD development, and it does not seem to worsen GERD when it was already present. Nevertheless, when the gastritis pattern is unknown before the antibiotic administration, the effect of H. pylori eradication on gastric acid secretion and the incidence of GERD is unpredictable. In the exceptional cases in which H. pylori eradication could have negative effects on GERD, its clinical relevance will be limited, and reflux symptoms or endoscopic esophagitis will favourably respond to the standard PPI antisecretory treatment. Therefore, again, when H. pylori eradication is indicated in a particular patient, the concomitant diagnosis of GERD should not change our attitude. Finally, is has recently been recommended to eradicate H. pylori infection in those patients with GERD needing long-term treatment with PPI, as some studies have reported that these drugs induce, in presence of the organism, an atrophic gastritis, with the consequent risk of gastric cancer. However, most of these studies have important methodological defects, and several authors have reported contrary results. In any case, the appearance in the gastric mucosa of clinically relevant lesions, such as intestinal metaplasia, dysplasia or adenocarcinoma, in patients treated with PPI for several years, has not yet been demonstrated, although this could simply be a problem of time. This question seems to be too controversial to be answered with the available data, and we should wait until new studies clarify this topic. In the meantime, as it occurs with any controversial indication, the decision of the doctor facing a patient infected by H. pylori and needing maintenance therapy with PPIs should be assessed on a case by case basis. PMID- 15899167 TI - [Fever after surgical procedure in a 54 year-old male]. PMID- 15899168 TI - [Restless legs syndrome and neuroacanthocytosis]. PMID- 15899169 TI - [Theophylline induced gynecomastia]. PMID- 15899170 TI - [Has the international normalized ratio (INR) been overestimated?]. PMID- 15899171 TI - [Safety of selective serotonin reuptake inhibitors]. PMID- 15899173 TI - [Role of integrons in antimicrobial resistance]. PMID- 15899174 TI - [Phenotypic and genotypic characteristics of Mycobacterium kansasii strains isolated in Spain (2000-2003)]. AB - INTRODUCTION: Mycobacterium kansasii is an opportunistic pathogen that mainly causes pulmonary infections. This species accounted for 9.7% of Mycobacteria other than tuberculosis complex identified in the reference laboratory of the Spanish Centro Nacional de Microbiologia during the period of 2000-2003. METHODS: In this study we analyzed the phenotypic and genotypic characteristics of 298 M. kansasii strains isolated over this 4-year period. The phenotypic characteristics were determined by conventional methods: biochemical testing, culture and morphological study. Genotypic characteristics were studied using PCR restriction fragment analysis of a fragment of the hsp65 gene and digestion with BstEII and HaeIII, according to the method of Telenti. RESULTS: Among the total of tested strains, 57.4% had the typical phenotypic characteristics described for M. kansasii. The rest had atypical patterns that we grouped into 17 biotypes. Strains belonging to six of the seven described genotypes were identified, with 86.6% of the strains falling into genotype I. CONCLUSION: Analysis of the phenotypic characteristics of M. kansasii showed a higher discrimination index for intraspecific differentiation than genotypic methods. Nevertheless, the high variability of phenotypic characteristics, some of which were very specific for the species (e.g., photochromogenicity), could complicate their identification. Hence both conventional and molecular methods should be used to accurately identify the atypical isolates. PMID- 15899175 TI - [Class 1 integrons among Salmonella enterica isolates producing different types of beta-lactamases from the health region of Tortosa (Spain)]. AB - OBJECTIVE: To assess the frequency of class 1 integrons among isolates of Salmonella enterica producing different types of beta-lactamases from the health region of Tortosa, and to attempt to establish the resistance genes located within their variable regions. METHODS: The presence of class 1 integrons and of aadA1, aadA2, dfrA1, tem-1, oxa-1 and pse-1 resistance genes within their variable regions was investigated by PCR in 100 ampicillin-resistant isolates of S. enterica (30 S. enteritidis, 56 S. Typhimurium and 14 from other serotypes) consecutively recovered in our laboratory between 2000 and 2001. Beta-lactamases were characterized by isoelectric focusing and PCR. RESULTS: a) 6/57 TEM-1 producing isolates carried integrons: 1 S. ser Panama, 2 S. ser Enteritidis and 1 S. ser Typhimurium (1600 pb/aadA1-dfrA1); 1 S. ser Panama (1600 pb/aadA2-dfrA1); 1 S. ser Grumpensis (1500 pb 1 1700 pb; aadA2 and ??) b) All OXA-1 producing isolates (20 S. ser Typhimurium) bore an integron of 2000 pb/aadA1-oxa-1; c) All PSE-1 producing isolates (22 S. ser Typhimurium, most of them 104 phage type, and 1 S. enterica immobile [4,12:-:-]) harbored 2 integrons (1000 pb/aadA1 and 1,00 pb/pse-1). CONCLUSION: The presence of class 1 integrons carrying oxa-1 or pse-1 resistance genes in all the OXA-1-producing and PSE-1-producing isolates investigated could have contributed to their spread and explain the increase in frequency of multiresistant S. ser Typhimurium isolates harboring these enzymes seen in the health region of Tortosa. In addition, we report the first isolate of S. ser enterica serotype Grumpensis harboring integrons. PMID- 15899176 TI - [Hydatidosis in the province of Salamanca (Spain): should we let down our guard?]. AB - INTRODUCTION: Hydatid disease is a major health problem in endemic countries. In Spain, this disease was notifiable from 1981 to 1996, although its incidence kept increasing until the end of this period. From 1997 on, hydatid disease is only surveyed by endemic autonomous regions. Thus, true incidences for human hydatid disease in Spain are currently unknown. In this study the cases recorded from 1996 to 2003 at the Hospital Universitario (Salamanca) which receives patients from all Salamanca province, were analyzed. Method. We obtained epidemiological and clinical data from patients at the University Hospital (reference hospital in Salamanca province) with registered CIE-10 codification 122.0 to 122.9 (hydatid disease), from January 1996 to December 2003, excluding those patients not arriving from Salamanca. We estimated the incidence of hydatid disesase in Salamanca province regarding the total population of the province. RESULTS: Our data show that the mean incidence (1996-2003) was 10,8/100.000 inhabitants, twice as many as previously reported by the notifiable diseases scheme. 30% of infected patients were younger than 45. In addition, during the last three years of study, several cases of paediatric hydatid disease were observed. The analysis of the origin of the cases showed a broad distribution throughout the whole province of Salamanca. CONCLUSIONS: The real incidence of this disease seems to be higher than previously estimated. Importantly, the several cases of paediatric hydatid disease detected within the last two years of our study, suggest a possible re emergence of hydatid disease. PMID- 15899177 TI - [Failure of levofloxacin therapy in two cases of community-acquired pneumonia caused by fluoroquinolone-resistant Streptococcus pneumoniae and complicated with empyema]. AB - BACKGROUND: Community acquired pneumonia (CAP) due to Streptococcus pneumoniae is a frequent cause of morbidity and mortality. We communicate two cases of CAP with complications. In both cases levofloxacin-resistant S. pneumoniae was isolated in pleural effusion. Patient 1: A 51-year-old man who had not received previous treatment with quinolones was admitted to the hospital for CAP and initially treated with levofloxacin (500 mg/24h iv). Four days later pleural effusion developed and fluid culture isolated levofloxacin-resistant S. pneumoniae (MIC > 32 .g/ml). The outcome was favorable following chest tube placement and treatment with beta-lactam antibiotics. Patient 2: A 73-year-old man with a history of chronic obstructive pulmonary disease was admitted due to CAP and was initially treated with levofloxacin (500 mg/24 h iv). He was transferred to our hospital after 10 days of treatment with this antibiotic, following the development of pleural effusion with isolation of levofloxacin-resistant S. pneumoniae (MIC = 12 .g/ml). The patient was treated with chest tube placement and beta-lactam antibiotics with a favorable outcome. CONCLUSIONS: Patients with CAP treated empirically must be closely followed, both clinically and radiologically, to facilitate early detection of complications due to bacterial resistance to the prescribed antibiotic. Patients with CAP who have received quinolones in the weeks before the development of pneumonia should not been treated empirically with these antibiotics because of the risk of resistance development. PMID- 15899178 TI - [Histoplasma capsulatum infection in a group of travelers to Guatemala]. AB - OBJECTIVE: To analyze the epidemiological and clinical characteristics of Histoplasma capsulatum infection in a group of volunteers who had traveled to Guatemala. METHODS: A review was performed of the clinical records of nine patients who consulted at the Tropical Medicine Outpatient Unit of Hospital Clinic in Barcelona, Spain. RESULTS. All patients were attended for travelers' diarrhea. In addition, one student had also presented with respiratory symptoms and fever while in Guatemala and her chest X-ray showed interstitial infiltrates. The epidemiological history recognized risk factors for histoplasmosis in all patients (all of them had been working in the reconstruction of an old school building). The intradermal histoplasmin test was performed and was positive in all nine patients. None of the students required treatment. CONCLUSIONS: Histoplasmosis should be included in the differential diagnosis of returning travelers' fever. The diagnosis would have been missed in all of our patients if not for a directed clinical history taking. It is important to reinforce prevention measures, e.g. the use of face masks, when working in an environment with a risk for acquiring histoplasmosis. PMID- 15899179 TI - [Paludism due to Plasmodium falciparum in visitors to the Dominican Republic]. AB - We present two cases of P. falciparum malaria in visitors to tourist resorts on the East Coast of the Dominican Republic, traditionally believed to be an area without risk of malaria. In both patients the malaria was severe (with 20% parasitization in one) and there was a long interval between the onset of symptoms and diagnosis. These cases are possibly related (along with a further 17 reports by the Centers for Disease Control and Prevention) to an increase in the population of Anopheles sp as a consequence of increased rainfall and floods provoked by a hurricane in September 2004, as well as to the presence of a semi immune population (Haitian immigrants working in the construction and tourist sectors). Both physicians and patients should be aware of this outbreak so that adequate precautions can be taken and early diagnoses can be made. PMID- 15899180 TI - [Recommendations of the CEVIHP/SEIP/AEP/PNS on antiretroviral treatment in HIV infected children and teenagers]. AB - OBJECTIVE: To update antiretroviral recommendations in antiretroviral therapy (ART) in HIV-infected children and adolescents. METHODS: Theses guidelines have been formulated by a panel of members of the Plan Nacional sobre el SIDA (PNS) and the Asociacion Espanola de Pediatria (AEP) by reviewing the current available evidence of efficacy, safety, and pharmacokinetics in pediatric studies. Three levels of evidence have been defined according to the source of data: Level A: randomized and controlled studies; Level B: Cohort and case-control studies; Level C: Descriptive studies and experts' opinion. RESULTS: When to start ART should be made on an individual basis, discussed with the family, considering the risk of progression according to age, CD4 and viral load, the ART-related complications and adherence. The ART goal is to reach a maximum and durable viral suppression. This is not always possible, even with clinical and immunologic improvement. The difficulties of permanent adherence and side-effects are resulting in a more conservative trend to initiate ART, and to less toxic and simpler strategies. Currently, combinations of at least three drugs are of first choice both in acute and chronic infection. They must include 2 NA 1 1 NN or 2 NA 1 1 PI. ART is recommended in all symptomatic patients and, with few exceptions, in all infants in the first year of life. Older asymptomatic children should start ART according to CD4 count, especially CD4 percentage, that vary with age. Despite potent salvage therapies, it is common not to reach viral undetectability. Therapeutical options when ART fails are scarce due to cross resistance. The cause of failure must be identified. Occasionally, there exists clinical and/or immunological progression, and a change of therapy with at least two new drugs still active for the patient, is warranted with the aim of increasing the CD4 count to a lower level of risk. Toxicity and adherence must be regularly monitored. Some aspects about post exposure prophylaxis and coinfection with HCV or HBV are discussed. CONCLUSIONS: A higher level of evidence with regard to ART effectiveness and toxicity in pediatrics is currently available, leading to a more conservative and individualized approach. Clinical symptoms and CD4 count are the main determinants to start and change ART. PMID- 15899181 TI - [Diseases produced by Bartonella]. AB - The genus Bartonella is cause of a broad number of emerging and re-emerging infectious diseases. Clinical manifestations depend on the implicated Bartonella sp. and the immunity of the host. Because there is not a universal therapy for this infection, treatment should be chosen individually. Bartonella sp. is responsible of potentially serious clinical pictures (endocarditis, chronic bacteremia, bacillary angiomatosis, Carrion's disease, etc.), so clinical suspicion, a quick diagnosis and an early treatment provide a better resolution. PMID- 15899182 TI - [Patient native of Ghana with diarrhea and proctalgia]. PMID- 15899183 TI - [Larvae in the external auditory canal and nasal fossae of an alcoholic patient]. PMID- 15899184 TI - [A two-year history of arthritis of the hand]. PMID- 15899185 TI - [Fever without an infectious focus and liver transplantation: Infection due to human herpes virus type 6 in transplanted patients]. PMID- 15899186 TI - [Spontaneous rupture of the spleen in a patient with visceral leishmaniasis]. PMID- 15899187 TI - [Sacroiliac abscess due to Serratia marcescens in an immunocompetent patient]. PMID- 15899188 TI - [Allergy to an antibiotic for flu treatment]. PMID- 15899189 TI - [Pulmonary arterial hypertension in adults with congenital heart disease]. AB - Pulmonary arterial hypertension is a chronic, persistent elevation in pulmonary artery pressure without evidence of left heart failure. Pulmonary hypertension is common in patients with adult congenital heart disease and is usually the result of an increase of pulmonary blood flow through a large left to right shunt. This condition is progressive and patients are symptomatic and usually die between the third and fifth decades of life. To date, there is no standardized treatment for this condition and a general policy of non-intervention to avoid destabilization of the balanced physiology is recommended. Intravenous prostanoids have been shown to have an effect but they are invasive and associated with major side effects. Lung and combined heart and lung transplantation might be a therapeutic option for selected patients. However, donor shortage is a major issue. Oral advanced therapies have been recently shown to improve haemodynamics and survival in idiopathic pulmonary hypertension or in pulmonary hypertension related to scleroderma and may have a role in patients with pulmonary hypertension secondary to congenital heart disease. PMID- 15899190 TI - [Tumor necrosis factor-alpha in heart failure: more questions than answers]. PMID- 15899191 TI - [Complexity and severity scores in cardiac surgery. Uses and limitations]. PMID- 15899192 TI - [Impaired endothelium-dependent forearm vasodilation in idiopathic dilated cardiomyopathy is related to severe left ventricular dysfunction and elevated serum tumor necrosis factor levels]. AB - INTRODUCTION AND OBJECTIVES: Endothelial dysfunction has been found in patients with idiopathic dilated cardiomyopathy (IDC), but its mechanism remains unknown. Our aim was to investigate whether forearm endothelium-dependent vasoreactivity correlates with cardiac disease severity or neurohormonal activation. PATIENTS AND METHOD: We studied 23 patients with IDC and 10 healthy sex- and age-matched controls using brachial artery ultrasound to assess flow-mediated dilation (FMD) and nitroglycerin-induced vasodilation (NIV). In the IDC group, we determined plasma neurohormone and cytokine levels at the same time. RESULTS: FMD was significantly less in the IDC group compared with the control group [--0.06 (2.8)% vs 4.4 (4.6)%, respectively; P<.01], whereas NIV was similar in both groups [15.0 (6.4)% vs 14.0 (7.4)%, respectively; P=NS]. FMD was significantly less in patients with poorer left ventricular (LV) function and more severe LV dilatation, and in those with a higher tumor necrosis factor-alpha (TNF-alpha) level. NIV was similar in all patient subgroups. There was a significant inverse correlation between the TNF-alpha plasma level and FMD (r=-0.75; P<.01). No correlation was found between the plasma levels of other neurohormones and FMD. CONCLUSIONS: FMD, but not NIV, was impaired in patients with IDC compared with control subjects. In patients, there were significant associations between FMD impairment and the severity of LV dilatation, the severity of LV systolic dysfunction, and the plasma TNF-alpha level. The strongest correlation was observed between TNF-alpha plasma level and FMD. These data suggest that TNF alpha may be implicated in endothelial dysfunction in patients with IDC. PMID- 15899193 TI - [In-hospital mortality and early preinfarction angina: a meta-analysis of published studies]. AB - INTRODUCTION AND OBJECTIVES: The occurrence of preinfarction angina (PA) reduces the extent of myocardial necrosis, increases the volume of viable myocardium, and improves left ventricular function. However, there is no agreement about the effect of PA on mortality. The objective of this study was to determine whether PA is associated with in-hospital mortality. METHOD: A meta-analysis (fixed effects model) of all published reports evaluating in-hospital mortality in patients with acute myocardial infarction according to the presence or absence of PA was performed. PA was defined as the occurrence of angina in the 24 hours before onset of the infarction. We searched the Medline and Embase databases in June 2004 using <> as search terms. Six studies involving a total of 3497 patients were finally identified. RESULTS: Only one study reported that PA had a statistically significant beneficial effect on in-hospital mortality. However, combining the data showed that the presence of PA was associated with a significant decrease in the probability of in-hospital death (odds ratio=0.61; 95% CI: 0.48-0.78; P<.0001. We did not detect any significant heterogeneity between the studies (chi2=5.92; P=.31). CONCLUSIONS: The occurrence of preinfarction angina in the 24 hours before the onset of myocardial infarction was associated with a significant reduction in in-hospital mortality of 39%. PMID- 15899194 TI - [Radiofrequency catheter ablation of ventricular tachycardia in patients with an implantable defibrillator. Long-term follow-up]. AB - INTRODUCTION AND OBJECTIVES: The frequent occurrence of ventricular tachycardia can be a serious problem for patients with an implantable defibrillator, and may necessitate adjuvant antiarrhythmic therapy or radiofrequency catheter ablation. We analyzed the long-term results obtained with this latter therapy in patients suffering from frequent or continuous ventricular tachycardia. PATIENTS AND METHOD: Eighteen ablation procedures were performed in 11 patients who had a defibrillator implanted because of previous syncopal ventricular tachycardia. All were men, aged 67.64 (5.87) years; 10 patients had had a myocardial infarction 15.50 (5.08) years earlier, and one suffered from arrhythmogenic right ventricular dysplasia. RESULTS: Electrophysiologically, treatment was initially successful in 8 patients (72.73%). After a follow-up period of 39.10 (24.70) months, the number of defibrillator discharges decreased significantly in all patients, from 52.82 (35.73) to 0.64 (1.03) (P=.001). During follow-up, ventricular tachycardia occurred in nine patients. In five, it took the same form as the ablated ventricular tachycardia. Six patients needed additional ablation procedures: two because of initial failure, three because of recurrence, and one because a different ventricular tachycardia occurred. In addition to the good electrophysiological results obtained, long-term clinical evolution was favorable in all patients. CONCLUSIONS: Radiofrequency ablation successfully disrupts frequent or continuous ventricular tachycardias and significantly reduces the defibrillator discharge rate even when ablation has failed electrophysiologically. It is particularly useful in these latter critical situations, in which other therapies are not sufficiently effective. Because our patients mainly had ischemic heart disease and were highly susceptible to new arrhythmias during follow-up, ablation complemented rather than replaced the implantable defibrillator. PMID- 15899195 TI - [Early heart rate increase does not predict the result of the head-up tilt test potentiated with nitroglycerin]. AB - INTRODUCTION AND OBJECTIVES: The magnitude of the change in heart rate during the first few minutes of the head-up tilt test has been used to predict the test's result. The aim of this study was to investigate whether the heart rate increase during the head-up tilt test potentiated with nitroglycerin is related to the development of syncope. PATIENTS AND METHOD: The study included 158 consecutive patients with syncope, with stable sinus rhythm, and without structural cardiac disease who were undergoing a head-up tilt test with nitroglycerin. The heart rate increment induced by the tilt maneuver and by nitroglycerin administration was calculated, and its relationship to clinical variables and to the test's results was analyzed. RESULTS: The head-up tilt test gave positive results in 117 patients (74%). The heart rate was 68.7 (11.3) bpm in the decubitus position and 85.1 (15.4) bpm during the first 6 min of tilting. There was strong inverse correlation between the heart rate increase induced by tilting and age (r=--0.63; P<.001), but the increase (16.8 [9.3] bpm in patients with syncope versus 14.9 [11.3] bpm in those without; P=.3) did not predict the result of the test. The heart rate increase induced by nitroglycerin was also similar for patients with and without syncope during the pharmacologic phase of the test (27.3 [12.6] bpm and 26.7 (13.4) bpm, respectively; P=.8). CONCLUSIONS: The magnitude of the heart rate increase during the first few minutes of tilt-testing and after nitroglycerin administration is inversely related to age but does not predict the result of the head-up tilt test with nitroglycerin. PMID- 15899196 TI - [Radial artery spasm in transradial cardiac catheterization. Assessment of factors related to its occurrence, and of its consequences during follow-up]. AB - INTRODUCTION AND OBJECTIVES: Radial artery spasm is the most frequent complication of transradial cardiac catheterization. It causes patient discomfort and reduces the procedure's success rate. The aims of this study were to identify variables associated with this complication, such as clinical parameters, angiographic characteristics of the radial artery and factors related to the procedure, and to analyze the clinical consequences of spasm, both generally and for radial artery patency, during follow-up. PATIENTS AND METHOD: The study included 637 patients who were undergoing transradial cardiac catheterization. Radial artery spasm was recorded using a scale that reflected the presence of pain and the technical difficulty of the procedure. RESULTS: Radial artery spasm was reported in 127 patients (20.2%). Multivariate analysis showed that the variables associated with radial artery spasm were radial artery anatomical anomalies (odds ratio [OR]=5.1; 95% confidence interval [95% CI]: 2.1-11.4), use of >size-3 catheters (OR=3.0; 95% CI: 1.9-4.7), moderate-to-severe pain during radial artery cannulation (OR=2.6; 95% CI: 1.4-4.9), the use of phentolamine as a spasmolytic (OR=1.8; 95% CI: 1.1-2.9), and postvasodilation radial artery diameter (OR=0.98; 95% CI: 0.98-0.99). At follow-up [20 (18) days], severe pain in the forearm was more frequent in patients who presented with radial artery spasm (12.4% vs 5.3%), but there was no significant difference in the radial artery occlusion rate (4.5% vs 2.2%). CONCLUSION: Radial artery spasm during transradial catheterization mainly depends on radial artery characteristics and procedural variables. At follow-up, radial artery spasm was associated with more frequent severe pain in the forearm, but the radial artery occlusion rate was not increased. PMID- 15899197 TI - [Trends in coronary artery bypass surgery: changes in the profile of the surgical patient]. AB - INTRODUCTION AND OBJECTIVES: In the last few years, the percentage of high-risk patients proceeding to coronary artery bypass surgery has increased. The most common risk factors are older age and the presence of comorbid complaints. We carried out a retrospective study to confirm this new risk profile and to evaluate its impact on surgical results. PATIENTS AND METHOD: We analyzed the changing risk profile of 1360 patients who underwent coronary artery bypass surgery in our hospital between 1993 and 2001, divided into three historical cohorts: 1993-1996, 1997-1999 and 2000-2001. The main factors associated with morbidity and mortality were analyzed by logistic regression analysis. The introduction of new operative techniques, such as off-pump surgery and arterial grafting, was also evaluated. RESULTS: The patients' risk profile worsened over time: patients were older, comorbid complaints were more common, and ventricular function was poorer. EuroSCORE figures reflected this trend: estimated mortality in the three historical cohorts was 2.0%, 4.0% and 4.2%, respectively (P<.001). However, risk-adjusted mortality, at 3.7%, 2.7% and 1.5%, respectively, decreased (P<.05), and combined overall morbidity and mortality remained stable, at 16.7%, 16.4% and 13.8%, respectively, (P<.39). There was a non-significant tendency for arterial grafting and off-pump surgery to reduce in-hospital morbidity and mortality. CONCLUSIONS: The risk profile of patients undergoing surgery has worsened as their mean age has increased and as comorbid complaints have become more prevalent. However, there has been no simultaneous increase in risk-adjusted mortality. The potential benefits of new surgical advances such as off-pump surgery and multiple arterial grafting must be corroborated by future studies. PMID- 15899198 TI - [Guidelines on diagnosis and treatment of pulmonary arterial hypertension]. PMID- 15899199 TI - [Percutaneous coronary intervention in myocardial infarction. Current concepts]. AB - Percutaneous mechanical reperfusion during acute myocardial infarction with ST segment elevation has proved to be the most effective way of quickly restoring adequate flow in the affected coronary artery. Randomized clinical trials have shown that percutaneous coronary intervention (PCI) is superior to thrombolysis. Initial fears about the use of stents in primary angioplasty vanished when clinical studies demonstrated that they gave better results than those obtained under optimal conditions with balloon angioplasty. The need to transfer patients to a cardiac catheterization laboratory for primary PCI does not decrease the efficacy of this form of treatment, which remains superior to immediate thrombolysis at the admitting hospital. Distal embolization can alter the situation by preventing myocardial reperfusion. Although there are many therapeutic strategies for managing thrombotic lesions, only early administration of glycoprotein IIb/IIIa inhibitors, direct stenting, and use of an X-Sizer device followed by stent implantation have been shown in randomized studies to lead to significant improvements in clinical or angiographic parameters. No technique has been shown to prevent damage due to myocardial reperfusion. However, it would be difficult to improve upon the good results achieved with PCI in the majority of patients. Rescue PCI is indicated when thrombolysis appears to have failed, especially when a catheterization laboratory is close by or when patients can be transferred early to a center with angioplasty facilities. For most cases of cardiogenic shock, PCI is the only therapeutic modality currently recommended. PMID- 15899200 TI - [Myocardial fibrosis in hypertrophic cardiomyopathy]. PMID- 15899201 TI - [Acute coronary events among spectators in a soccer stadium]. AB - The aim of this study was to describe acute coronary syndromes (ACS) with a substantial emotional component in spectators of professional league competition sports events. The study was done at the Barcelona soccer team's home stadium. We recorded cases of ACS that occurred during official competition matches during the 2000-2001 season. A total of 7 episodes of ACS were recorded, 1 involving sudden death, 4 involving acute myocardial infarction and 2 involving angina pectoris. The victim of sudden death had a history of heart disease. The overall risk of ACS was 0.0056 episodes per 100,000 person-hours. We conclude that medical facilities at the stadium facilitated the initial diagnosis of ACS and ensured prompt initial treatment and transport to the reference hospital. PMID- 15899202 TI - [Ductus arteriosus patency with stenting in critical pulmonary stenosis and pulmonary atresia with intact interventricular septum]. AB - We report our experience with stenting of the ductus arteriosus in three neonates. Two patients had pulmonary atresia with an intact interventricular septum and one had critical pulmonary stenosis. Radiofrequency ablation was used to open the atretic pulmonary valve in two patients. In all three patients implanting a stent avoided the need for surgical creation of a Blalock-Taussig shunt. In two patients the procedure was scheduled as elective surgery, and in one it was done as an emergency procedure. Ductus arteriosus stenting is an alternative to palliative cardiac surgery. PMID- 15899203 TI - [Aspergillus fumigatus endocarditis in a patient with a biventricular pacemaker]. AB - Aspergillus fumigatus endocarditis is one of the rarest and severest complications in cardiological patients. We describe a patient with an intracardial pacemaker who was diagnosed as having Aspergillus fumigatus endocarditis. Postmortem examination showed a large, Aspergillus-infected thrombus encased in the right ventricle, pulmonary trunk and main pulmonary branches. PMID- 15899204 TI - [Acute myocardial ischemia and ventricular thrombus associated with pheochromocytoma]. AB - We describe a patient with a suprarenal pheochromocytoma that had a complex course with electrocardiographic findings characteristic of diffuse myocardial damage, normal findings on coronary angiography, and left intraventricular thrombus complicated by embolic stroke. PMID- 15899205 TI - [Duplicate or redundant publication: can we afford it?]. PMID- 15899206 TI - [Prophylaxis for hypertrophic cardiomyopathy and infective endocarditis: from recommendations to implementation]. PMID- 15899207 TI - Tobacco-related cancer: two murders and one victim. PMID- 15899208 TI - [Delays and treatment interruptions: difficulties in administering radiotherapy in an ideal time-period]. AB - Prescribed total radiation dose should be administered within in a specific time frame and delays in commencing treatment and/or unplanned interruptions in radiation delivery are unacceptable because, in certain cancer sites, treatment time prolongation can have a deleterious effect on local tumour control, and on patient outcomes. The present review evaluated the causes of initial treatment delays as well as interruptions in the scheduled radiotherapy. The literature search highlighted a significant concern in avoiding treatment-time prolongation in head and neck, cervix, breast and lung cancer. Among the causes involved in delay in radiotherapy commencement factors such as waiting lists, lack of material and human resources, and an increase complexity in planning, simulation and verification are highlighted. Most authors recommend radiotherapy commencement as soon as possible in radical (exclusive irradiation with active tumour present) and palliative situations with a maximum delay of no more than 6 to 8 weeks in the case of adjuvant radiotherapy (post-resection) programs. Interruptions during the course of treatment include: planned unit maintenance and servicing, acute patient toxicity or unexpected malfunction of linear accelerators; this last feature has the most deleterious effect on patients as well as radiotherapy practitioners. Interruptions that impact on the programmed time-course for radiotherapy needs to be compensated-for so as assure the biological equivalence in treatment efficacy with respect to cancer site and stage. PMID- 15899209 TI - [Extra-pulmonary small-cell cancer: is its outcome similar to small cell lung cancer?]. AB - INTRODUCTION: Extra-pulmonary small cell carcinoma (ESCC) is as a pathologic entity distinct from small cell lung carcinoma (SCLC). ESCC is considered a systemic disease in its origin, so the therapeutic approach is similar to SCLC with chemotherapy being considered in case of extensive and local disease. We present a retrospective comparison of ESCC and SCLC in our institution. MATERIAL AND METHODS: Using the tumour registry database of Hospital Universitario La Fe we reviewed 24 ESCC cases receiving attention between 1987 and 2003, and these were compared with a series of 341 patients with SCLC in the same institution. RESULTS: Of the 24 patients with ESCC 19 were men and 5 were women with an average age of 58 years (range 23 to 85). The most frequent site was the mediastinum with 58% having extensive disease. All patients but one received treatment. The therapeutic approaches were local and systemic in 13 patients, systemic alone in 6 and local alone in 4. Schedules based on platinum and etoposide were used. The median follow-up was 53 months (range 4 to 211). Median survival was 18.9 months; 30 in patients with local disease and 8 in those with extensive disease. In the SCLC series of patients, there were 336 men and 5 women; 62% having extensive disease. The median survival was 10 months; 12 months in those patients with local disease and 8 in those with extensive disease. CONCLUSIONS: The overall survival of patients with ESCC was slightly better than patients with SCLC. ESCC with local disease had a better survival outcome than SCLC with local disease. Chemotherapy is the cornerstone of the treatment, but sometimes local treatment could be sufficient. PMID- 15899210 TI - Concomitant boost radiation and concurrent cisplatin for advanced head and neck carcinomas. Preliminary results of a phase II, single-institutional trial. AB - INTRODUCTION: This study aims to asses the effectiveness and toxicity of boost radiotherapy concomitant and concurrent cisplatin for patients with locally advanced head and neck cancer (LAHNC). MATERIAL AND METHODS: There were 30 patients included in a prospective, phase II single-institution trial and of whom, 29 were at AJCC stage IV and 1 at stage III. Treatment consisted of radiotherapy acceleration fractionation with concomitant boost, 72 Gy, and 2 cycles of concomitant cisplatin (20 mg/m2/day continuous infusion; days 1-5 and 29-33). Amifostine, (i.v. 200 mg/m2) was administered to 26 prior to the first fraction of radiotherapy. Endpoints of the study were quality-of-life (QL), overall survival, and local control of disease. RESULTS: Complete response (CR) was achieved in 23 patients (77%), 2 patients had partial response (PR) (7%), 4 had no response (13%), and 1 was not evaluated for response. The 2-year overall survival and loco-regional control were 60% and 56%, respectively. Main toxicity was grade 3 or 4 mucositis in 93% of the patients. QL scores (questionnaire QLQC30; version 3.0) and the HN cancer module QLQ-HN35) showed a worsening in areas related to the treatment e.g. dry mouth, problems stretching the mouth, and sticky saliva. CONCLUSIONS: this combination modality is active, but toxic, in the treatment for LAHNC. Concomitant boost radiotherapy is probably, not the best radiotherapy schema for combining with chemotherapy in LAHNC. PMID- 15899211 TI - A phase II study of vinorelbine and estramustine in patients with hormone resistant prostate cancer. AB - INTRODUCTION: This phase II study was designed to evaluate the efficacy of vinorelbine in combination with estramustine in patients with chemotherapy-naive hormone-refractory prostate cancer. MATERIAL AND METHODS: Patients received vinorelbine (i.v. 25 mg/m2) on days 1 and 8 every 3 weeks and estramustine (oral, 600 mg/m2) daily. Eligible patients were required to have progressive metastatic disease following the first hormonal manipulation. RESULTS: Of the 51 patients enrolled (median age = 69 years), 84% presented bone involvement and 75% had at least two organs involved at the time of study entry and 47 were evaluable for treatment efficacy. Prostate specific antigen (PSA) response (> or =50% decrease) which was the primary efficacy criterion was reported in 21 patients (41.2%) in the intent-to-treat (ITT) population and in 20 patients (48.8%) in the per protocol (PP) population. Of the 7 patients with measurable disease, 2 achieved partial response. Median progression-free survival and overall survival were 4.7 months (range: 1.9-8.6) and 14.3 months (range: 4.2-21.2), respectively. Grade 3 4 neutropenia was reported in 6.1% of patients and in 1% of cycles. The incidence of complicated neutropenia (febrile neutropenia reported in 1 patient and septic shock with severe neutropenia reported in 2 patients) was 5.8%. The most frequent grade 3-4 non-haematological events (% of patients > or =5%) included anorexia (10%), thrombosis/embolism (8%), vomiting and hypotension (6% each). There were 3 toxic deaths (5.9 %) resulting from pulmonary embolism, angina pectoris, and septic shock. The impact of combined chemotherapy on the quality-of-life (QL) of the patients was assessed between baseline and the first evaluation scheduled at 6 weeks indicated a marked reduction in pain while the rest of the symptoms remained stable. Overall, health status improved slightly over the treatment period. CONCLUSIONS: This study confirmed that the combination of vinorelbine and estramustine is an active regimen in patients with hormone-resistant prostate cancer who had not been treated previously with chemotherapy. Main toxicities included complicated neutropenia even though the incidence of severe neutropenia was low. We observed a higher incidence of toxic deaths which could have been related to the regimen of estramustine used in the study. PMID- 15899212 TI - [Dyspnea, oliguria and lower extremities edema in a patient undergoing mediastinic radiotherapy for a non-resectable thymic carcinoma]. AB - In this case report we present the clinical signs and symptoms of cardiac tamponade of tumor origin. We explain the clinical investigations for its diagnosis as well as its specific differential diagnosis in cancer patients. The different therapeutic options are also presented together with a brief summary of the thymic carcinoma. PMID- 15899213 TI - [Primary linitis plastica of the rectum]. AB - Primary linitis plastica of the rectum is an infrequent and unusual rectal carcinoma. We report a new case that presented clinically as a haemorrhoidal syndrome. We comment on the characteristics--clinical picture and histopathology- of this uncommon presentation of rectal tumour, as well as its clinical history and prognosis taking into account that the literature consulted indicated the unusualness of primary linitis plastica of the rectum. PMID- 15899214 TI - Wilms tumor: what's new? PMID- 15899215 TI - Neutropenia and enterocolitis with docetaxel treatment. PMID- 15899216 TI - Integrating trastuzumab in the treatment of breast cancer. Current status and future trends. PMID- 15899217 TI - [Current status of the prognostic value of molecular markers in patients with colorectal cancer and the prediction of response to adjuvant therapy]. AB - Colorectal cancer is one of the best studied of all malignant diseases in terms of genetics and/or molecular prognostic factors. These factors, and relationships with prognosis, may have important implications especially in the design of surgical and adjuvant chemo-radiotherapy options. However, the true prognostic significance of all known factors has yet to be realised. We have reviewed the literature with specific focus on the role of molecular markers involved in prognosis and the prediction of response to adjuvant treatment. PMID- 15899218 TI - Immunoexpression of p53, Ki-67 and E-cadherin in basaloid squamous cell carcinoma of the larynx. AB - INTRODUCTION: Basaloid is a rare and poorly-differentiated variant of squamous cell carcinoma of the larynx, with an invasive solid growth of cells in a lobular configuration. Different molecular markers, such as p53, Ki-67 and E-cadherin, have been shown to be prognostic factors in head and neck cancer. OBJECTIVE: To evaluate the relationship between the immunoexpression of p53, Ki-67 and E cadherin in relation to prognosis in basaloid squamous cell carcinoma of the larynx (BSCCL). PATIENTS AND METHODS: We retrospectively studied 11 cases of BSCCL, all male with a mean age of 62.4 years. Immunohistochemical analyses were performed on paraffin-embedded tissues using p53 (DO- 7), Ki-67 (MIB-1) and E cadherin (36B5) antibodies. Quantitative assessments of the expression and descriptive statistical analyses were performed. RESULTS: In 72.7% of the cases, clinically advanced stages III-IV were diagnosed. Average survival time was 56.09 months, and 72.7% of patients died as a consequence of the tumour. Immunoreactivity of p53 (>10% of cells) was detected in the 81.8% of the cases. The 72.7% of the cases showed overexpression of Ki-67 (>50% of cells). The cases with low immunoexpression of Ki-67 and p53 had the best clinicopathological data. All cases showed a decreased expression of E-cadherin. CONCLUSIONS: BSCCL is an aggressive variant of the squamous cell carcinoma and has a high expression of p53 and Ki-67 with a low expression of Ecadherin. These results could be related to the aggressiveness of the disease and its poor prognosis. PMID- 15899219 TI - [Experimental model for treating pulmonary metastatic melanoma using grape-seed extract, red wine and ethanol]. AB - INTRODUCTION: Melanoma is one of the neoplasias that most frequently metastasize, especially in the lung, where represents a challenge in oncology since current treatment is ineffective, and mortality is high. MATERIAL AND METHODS: Swiss mice (n = 52) were inoculated with 0.5 x 106 B16F10 cell lines and, later, given an oral administration of grape-seed extract, red wine or ethanol. Metastatic nodules on the lung surface were counted and, after processing for microscopy, five sections were selected for image analysis and the invasion index was calculated. RESULTS: Macroscopic analysis showed that grape-seed extract and red wine reduced the number of metastatic nodules by 26.07 and 20.81%, respectively, compared with a control group treated with ethanol. Microscopically, the reduction in the invasion index was 31.65 for grape-seed extract and 17.57% for red wine. CONCLUSION: Ethanol administration significantly increased pulmonary metastasis while grape-seed extract and red wine led to their reduction. PMID- 15899220 TI - [Analysis of tumour-derived DNA in serum in advanced non-microcytic lung cancer: might it be a prognostic factor?]. AB - INTRODUCTION: Presence of circulating DNA in the serum of patients with cancer makes detection of tumour-specific genetic alterations feasible. OBJECTIVE: To study serum DNA concentration in patients diagnosed as having advanced Non-Small Cell Lung Cancer (NSCLC) and to evaluate its relationship with age, histology, stage, response, time-to-progression (TTP), and survival. METHODS: Serum DNA from 78 patients was purified and spectrophotometrically quantified. RESULTS: No significant correlations were found between serum DNA concentration and age, histology, response and survival. There was a significant correlation with respect to stage (IIIB = 408.75 ng/ml; IV = 478.74 ng/ml; p = 0.02). When patients were grouped according to DNA concentration, significant correlation with TTP was found; establishing a cut-off point at 500 ng/ml ([DNA] < 500 ng/ml TTP = 7.25 months, 95%CI: 3.5-5.25; [DNA ] > or = 500 ng/ml TTP = 4.25 months, 95%CI: 2-6.5; p = 0.05). CONCLUSIONS: Using the present method, DNA concentration quantification appears to be simple, but with certain deficiencies due to inter sample variability and low specificity. This is because total DNA concentration is measured without distinguishing as to whether it is tumour-related. We suggest that there is a correlation between DNA concentration and prognosis which enables an analysis of the natural history of the disease. PMID- 15899221 TI - [Extra-cranial anaplastic oligoastrocytoma development from a low-grade glioma]. AB - Oligodendrogliomas are primary brain tumours derived from oligodendroglial cells, or precursors, and represent 2%-5% of brain tumours. This type of glioma has a favourable prognosis compared to other brain tumours. The treatment is multidisciplinary and is based on three therapeutic arms: surgery, radiotherapy and chemotherapy. We present a patient who had received treatment previously for a lowgrade glioma and who subsequently developed an anaplastic oligoastrocytoma in the same zone together with skull and extra-cranial involvement in the disease progression. PMID- 15899222 TI - [Extensive surgical resection in the treatment of invasive thymoma]. AB - In 10%-15% of patients, myasthenia gravis is associated with thymoma. Because of its site, this tumour can have an un-noticed clinical history over a considerable time and be diagnosed when adjacent structures such as mediastinal vessels, heart or trachea become infiltrated. We present a patient with thymoma and infiltration of the superior vena cava, and which represents an incipient vena cava syndrome. The surgical treatment consisted of complete resection of the tumour including partial replacement of the vena cava with a pericardial patch. The surgery was complemented with radiotherapy. PMID- 15899223 TI - [Neuroblastoma]. PMID- 15899224 TI - [Connect: the World of Critical Care Nursing]. PMID- 15899225 TI - ['Intuition or exactness? Intuitive positioning of the bed headboard in critical patients. Do we need to measure it?]. AB - OBJECTIVE: To explore the skill of the nursery staff of an intensive care unit (ICU) to set the head of the bed accurately and to determine the need to incorporate a slope indicator in each bed. METHODS: Observational prospective study to measure the positioning of the head of the bed at 30 degrees and 45 degrees from 0 degrees in beds with slope indicator that was hidden during the study. SETTING: A neurosurgery ICU with thirteen boxes. STUDY PERIOD: from November-2003 to January-2004. Subject : nursery staff. RESULTS: 228 measures were made, 160 (50% at 30 degrees and 50% at 45 degrees ) were made by nurses, and 68 (50% at 30 degrees and 50% at 45 degrees ) by auxiliary nurses. At both positions there were 26 correct settings. Mean was 20 degrees and 30 degrees at 30 degrees and 45 degrees , respectively. There were not differences between nurses and auxiliary nurses at 30 degrees (22.6 +/- 1.5 vs 21.8 +/- 2.4) nor at (33.3 +/- 2 vs 33.8 +/- 3.3). There were not significant differences due to work experience nor respect of analysed variables (intracranial pressure monitoring, squatting, etc.). CONCLUSION: We set the head of the bed under the recommended slope for this type of patients, regardless of the analysed variables. We think that an objective slope degree indicator is mandatory in this kind of beds in order to achieve an accuracy positioning. PMID- 15899226 TI - [Intensive care nursing discharge reports in Spain: present situation and analysis]. AB - OBJECTIVES: To determine the ICUs of Spain that fill out nursing discharge report, analyze what information they supply and know how the information on the patients is provided by the ICUs that do not use this record. MATERIAL AND METHODS: Observational descriptive study. The nursing discharge reports issued by the ICUs were analyzed. For the units that did not fill them out, it was documented how they provided information on the patient to the ward. RESULTS: A total of 259 ICU (83%) responded and 176 (68%) filled out the nursing discharge report, which was sent with the patient's history in all the units. A total of 99% of the reports included information on the eating and elimination needs and 7% on living according to beliefs in their structure. One hundred percent requested information on the perception/control and nutritional/metabolic patterns; 1% on sexuality/reproduction. Forty three (54%) of the 79 ICU (30%) that did not fill out a nursing discharge report accompanied the patient to the ward and gave an oral report. In all, 242 ICU (93%) provided information to the ward professionals added to that contained in the clinical history. CONCLUSIONS: a) the nursing professionals in a significant number of intensive care units filled out a discharge report when the patient was transferred to the ward; b) assessment of basic biological aspects prevailed in these reports over psychological and socioculture aspects; c) the units that did not fill out a discharge report accompany the patient to the ward or give a report by telephone of the patient's situations to the destination ward; and d) most of the ICUs guarantee the continuity of nursing cares when the patient is discharged, either by filling out a nursing discharge report, accompanying the patient to the ward or calling by telephone to the ward's professionals. PMID- 15899227 TI - [Organization of visits in Spanish ICU]. AB - It is a fact that visits in the intensive care units (ICU) have always been very restrictive due to diverse criteria. Nowadays, some of them have been progressively liberalized, giving to the relatives more satisfaction and a better cover to their necessities. In our country there are no surveys about the organization of these visits at the present time, apart from some exceptional cases which commentate the situation in a particular center. OBJECTIVE: Know how are visits organized in the intensive cares units in Spain. MATERIAL AND METHOD: DESIGN: cross-sectional study; scope: 98 ICU of different Spanish hospitals; studied variables: the number of visits, their duration, the number of relatives in each visit, the number of relatives visiting the patients at the same time, the amount of information each day, the professional that informs, the use of written information and information by telephone, preventive measures taken before the entry in the unit, how relatives take part in the cares, location of the waiting room and use or not of a protocol of attention to relatives. INSTRUMENTATION: The information has been obtained by a questionnaire placed in a web site of the Internet and directed to professionals of different Intensive cares units. RESULTS: The ninety and eight evaluated units represent 37 Spanish provinces from all the autonomous communities. The most frequently noticed pattern for the organization of visits shows a restrictive character of the visits in Spain. CONCLUSIONS: There are clear differences in the organization of visits between the studied ICU and no connection between the most common practice and what the survey, after evaluating the necessities of the relatives, recommends. These recommendations are: more visits, more flexibility of schedules, more integration of the relatives in the patients cares, broach the information from diverse disciplines, washing hands as the only necessary measure to be taken by the relatives. PMID- 15899228 TI - [Review of knowledge on care of patients with cardiovascular problems]. PMID- 15899229 TI - [Review of knowledge on the care of patients with endocrine-metabolic problems]. PMID- 15899230 TI - [Physiology of pancreatic secretion]. PMID- 15899231 TI - [Introduction. Exocrine pancreatic insufficiency. How does it happen? When and how should it be diagnosed? How should it be treated?]. PMID- 15899232 TI - [Study of the exocrine pancreatic function]. PMID- 15899233 TI - [Physiopathology, diagnosis, and treatment of exocrine pancreatic insufficiency after acute pancreatitis]. PMID- 15899234 TI - [Physiopathology, diagnosis, and treatment of exocrine pancreatic insufficiency in patients with chronic pancreatitis]. PMID- 15899235 TI - [Physiopathology, diagnosis, and treatment of exocrine pancreatic insufficiency in patients with cystic fibrosis]. PMID- 15899236 TI - [Physiopathology, diagnosis, and treatment of exocrine pancreatic insufficiency in patients with pancreatic cancer]. PMID- 15899237 TI - [Physiopathology, diagnosis, and treatment of exocrine pancreatic insufficiency in patients following gastrointestinal or pancreatic surgery]. PMID- 15899238 TI - [Physiopathology, diagnosis, and treatment of exocrine pancreatic insufficiency in other clinical situations: diabetes mellitus and HIV infection]. PMID- 15899239 TI - Transplantation of GABAergic neurons into adult mouse neocortex. AB - GABAergic neurons in the neocortex contribute to various brain functions by regulating cortical pyramidal neurons. A deficiency of GABAergic neurons in the neocortex leads to the dysregulation of cortical neuronal circuits, but this can be overcome by cell transplantation, which provides a practical approach to repair damaged neuronal circuits. Here, we focused on the transplantation of committed neuronal progenitor cells. Because neuronal differentiation is considerably suppressed in the adult neocortex, we transfected proneural bHLH transcription factors into neural precursor cells to commit them to a neuronal lineage prior to the cell transplantation. We show that ventral neural stem cells transfected with Ngn1 are integrated as GABAergic neurons within a few days of transplantation into the adult mouse neocortex. These results demonstrate that the transplantation of committed neuronal progenitor cells is an effective method for brain repair. PMID- 15899240 TI - Human adult olfactory neural progenitors rescue axotomized rodent rubrospinal neurons and promote functional recovery. AB - Previously, our lab reported the isolation of patient-specific neurosphere forming progenitor lines from human adult olfactory epithelium from cadavers as well as patients undergoing nasal sinus surgery. RT-PCR and ELISA demonstrated that the neurosphere-forming cells (NSFCs) produced BDNF. Since rubrospinal tract (RST) neurons have been shown to respond to exogenous BNDF, it was hypothesized that if the NSFCs remained viable following engraftment into traumatized spinal cord, they would rescue axotomized RS neurons from retrograde cell atrophy and promote functional recovery. One week after a partial cervical hemisection, GFP labeled NSFCs suspended in Matrigel matrix or Matrigel matrix alone was injected into the lesion site. GFP-labeled cells survived up to 12 weeks in the lesion cavity or migrated within the ipsilateral white matter; the apparent number and mean somal area of fluorogold (FG)-labeled axotomized RST neurons were greater in the NSFC-engrafted rats than in lesion controls. Twelve weeks after engraftment, retrograde tracing with FG revealed that some RST neurons regenerated axons 4-5 segments caudal to the engraftment site; anterograde tracing with biotinylated dextran amine confirmed regeneration of RST axons through the transplants within the white matter for 3-6 segments caudal to the grafts. A few RST axons terminated in gray matter close to motoneurons. Matrix alone did not elicit regeneration. Behavioral analysis revealed that NSFC-engrafted rats displayed better performance during spontaneous vertical exploration and horizontal rope walking than lesion Matrigel only controls 11 weeks post transplantation. These results emphasize the unique potential of human olfactory neuroepithelial-derived progenitors as an autologous source of stem cells for spinal cord repair. PMID- 15899241 TI - Effect of apolipoprotein E deficiency on reactive sprouting in the dentate gyrus of the hippocampus following entorhinal cortex lesion: role of the astroglial response. AB - This study investigated the effect of apolipoporotein E (apoE) deficiency on hippocampal reactive sprouting responses of the septohippocampal cholinergic (SHC) and commissural/associational fibers (C/A) following an electrolytic lesion of the entorhinal cortex (ECL), using apoE knockout (apoEKO) and age-matched control wild-type mice. Based on recent evidence suggesting that apoE plays a role in the modulation of glial inflammation, we also tested the hypothesis that the pattern of the astroglial response to ECL might be related to the defective reinnervation previously reported in apoEKO mice. Consistent with our hypothesis, we report a differential pattern of astroglial response that concurred with impairments in the sprouting of the SHC and corresponding synaptic replacement in apoEKO mice at 14 and 30 days post-lesion (DPL), a time range covering the onset of axonal/terminal sprouting to synaptogenesis. We also report a limited sprouting of the C/A fiber system in apoEKO relative to control mice at 30 DPL, a period of active dendritic remodeling. The results of the present study confirm and extend previous findings that apoEKO mice display impaired regenerative capacity in response to ECL and argue that in addition to the effect of apoE on lipid trafficking, apoE may also influence the astroglial response to damage, and that both of these effects account for the defective reinnervation observed in apoEKO mice. PMID- 15899242 TI - Histamine H(1) and H(3) receptors in the rat thalamus and their modulation after systemic kainic acid administration. AB - In rat thalamus, histamine H(1) receptor and isoforms of H(3) receptor were expressed predominantly in the midline and intralaminar areas. Correspondingly, higher H(1) and H(3) receptor binding was also detected in these areas. All isoforms of H(3) receptor were expressed in several thalamic nuclei, but there were minor differences between their expression patterns. H(1) mRNA expression was high in the ventral thalamus, but the H(1) binding level was low in these areas. Since increased brain histamine appears to have an antiepileptic effect through the H(1) receptor activity, kainic acid (KA)-induced status epilepticus in rat was used to study modulation of H(1) and H(3) receptors in the thalamus following seizures. After systemic KA administration, transient decreases in mRNA expression of H(1) receptor and H(3) receptor isoforms with full-length third intracellular loops were seen in the midline areas and the H(1) receptor mRNA expression also decreased in the ventral thalamus. After 1 week, a robust increase in mRNA expression of H(3) receptor isoforms with a full-length third intracellular loop was found in the ventral posterior, posterior, and geniculate nuclei. The changes indicate a modulatory role of H(3) receptor in the sensory and motor relays, and might be involved in possible neuroprotective and compensatory mechanisms after KA administration. However, short-term increases in the H(3) receptor binding appeared earlier (72 h) than the increases of H(3) mRNA expression (1-4 w). The elevations in H(3) binding were evident in the intralaminar area, laterodorsal, lateral posterior, posterior and geniculate nuclei, and were likely to be related to the cortical and subcortical inputs to thalamus. PMID- 15899243 TI - Activation of c-Jun and ATF-2 in primate motor cranial nerve nuclei is not associated with apoptosis following axotomy. AB - Nerve transection induces complex changes in gene regulation and expression that can have profound phenotypic effects on the fate of axotomized neurons. The transcription factors c-Jun and ATF-2 (activating transcription factor-2) are components of a regulatory network that mediates survival, regeneration, and apoptosis following axotomy in rodents. The activation and function of c-Jun and ATF-2 after nerve injury have not been examined in primates. Using a novel model of cranial nerve injury in baboons, we have examined the temporality of c-Jun activation (phosphorylation) in cranial nerve (CN) III and CN VI neurons and ATF 2 activation in CN VI neurons at 2, 4, and 9 days post-injury by immunohistochemistry. Furthermore, we have addressed whether the activation of these factors is associated with apoptosis by the TUNEL assay. We report that activated c-Jun is present in CN III and CN VI neurons ipsilateral to axotomy at 2, 4, and 9 days post-injury, but not in neurons contralateral to injury. Additionally, CN VI neurons ipsilateral to injury at 4 and 9 days contain activated ATF-2. Furthermore, no evidence of TUNEL reactivity was observed in either nucleus, regardless of laterality, at any of the examined time points. These findings suggest that activation of both c-Jun and ATF-2 does not mediate apoptosis in axotomized primate CN III and CN VI neurons at time points up to 9 days. This report serves as a basic inquiry into the neuronal response to cranial nerve injury in primates. PMID- 15899244 TI - Pharmacological validation of a mouse model of l-DOPA-induced dyskinesia. AB - Dyskinesia (abnormal involuntary movements) is a common complication of l-DOPA pharmacotherapy in Parkinson's disease, and is thought to depend on abnormal cell signaling in the basal ganglia. Dopamine (DA) denervated mice can exhibit behavioral and cellular signs of dyskinesia when they are treated with l-DOPA, but the clinical relevance of this animal model remains to be established. In this study, we have examined the pharmacological profile of l-DOPA-induced abnormal involuntary movements (AIMs) in the mouse. C57BL/6 mice sustained unilateral injections of 6-hydroxydopamine (6-OHDA) in the striatum. The animals were treated chronically with daily doses of l-DOPA that were sufficient to ameliorate akinetic features without inducing overt signs of dyskinesia upon their first administration. In parallel, other groups of mice were treated with antiparkinsonian agents that do not induce dyskinesia when administered de novo, that is, the D2/D3 agonist ropinirole, and the adenosine A2a antagonist KW-6002. During 3 weeks of treatment, l-DOPA-treated mice developed AIMs affecting the head, trunk and forelimb on the side contralateral to the lesion. These movements were not expressed by animals treated with ropinirole or KW-6002 at doses that improved forelimb akinesia. The severity of l-DOPA-induced rodent AIMs was significantly reduced by the acute administration of compounds that have been shown to alleviate l-DOPA-induced dyskinesia both in parkinsonian patients and in rat and monkey models of Parkinson's disease (amantadine, -47%; buspirone, -46%; riluzole, -33%). The present data indicate that the mouse AIMs are indeed a functional equivalent of l-DOPA-induced dyskinesia. PMID- 15899245 TI - Epileptogenesis and chronic seizures in a mouse model of temporal lobe epilepsy are associated with distinct EEG patterns and selective neurochemical alterations in the contralateral hippocampus. AB - Major aspects of temporal lobe epilepsy (TLE) can be reproduced in mice following a unilateral injection of kainic acid into the dorsal hippocampus. This treatment induces a non-convulsive status epilepticus and acute lesion of CA1, CA3c and hilar neurons, followed by a latent phase with ongoing ipsilateral neuronal degeneration. Spontaneous focal seizures mark the onset of the chronic phase. In striking contrast, the ventral hippocampus and the contralateral side remain structurally unaffected and seizure-free. In this study, functional and neurochemical alterations of the contralateral side were studied to find candidate mechanisms underlying the lack of a mirror focus in this model of TLE. A quantitative analysis of simultaneous, bilateral EEG recordings revealed a significant decrease of theta oscillations ipsilaterally during the latent phase and bilaterally during the chronic phase. Furthermore, the synchronization of bilateral activity, which is very high in control, was strongly reduced already during the latent phase and the decrease was independent of recurrent seizures. Immunohistochemical analysis performed in the contralateral hippocampus of kainate-treated mice revealed reduced calbindin-labeling of CA1 pyramidal cells; down-regulation of CCK-8 and up-regulation of NPY-labeling in mossy fibers; and a redistribution of galanin immunoreactivity. These changes collectively might limit neuronal excitability in CA1 and dentate gyrus, as well as glutamate release from mossy fiber terminals. Although these functional and neurochemical alterations might not be causally related, they likely reflect long-ranging network alterations underlying the independent evolution of the two hippocampal formations during the development of an epileptic focus in this model of TLE. PMID- 15899246 TI - Decreased levels of BDNF protein in Alzheimer temporal cortex are independent of BDNF polymorphisms. AB - Levels of brain-derived neurotrophic factor (BDNF) are reduced in specific brain regions in Alzheimer's disease (AD) and BDNF gene polymorphisms have been suggested to influence AD risk, hippocampal function, and memory. We investigated whether the polymorphisms at the BDNF 196 and 270 loci were associated with AD in a clinical and neuropathological cohort of 116 AD cases and 77 control subjects. To determine how BDNF protein levels relate to BDNF polymorphisms and AD pathology, we also measured BDNF in temporal association cortex, frontal association cortex, and cerebellum in 57 of the AD and 21 control cases. BDNF protein levels in temporal neocortex of the AD brains were reduced by 33% compared to control brains, whereas levels were unchanged in frontal and cerebellar cortex. The BDNF genotypes were not significantly associated with a diagnosis of AD, although the BDNF 270 C allele was slightly overrepresented among carriers of the APOEepsilon4 allele. Moreover, BDNF protein levels did not differ between the various BDNF genotypes and alleles. Neuropathologically, the loss of BDNF in AD showed a weak correlation with accumulation of neuritic amyloid plaques and loss of the neuronal/synaptic marker synaptophysin. The results suggest that the investigated BDNF polymorphisms are neither robust genetic risk factors nor determinants of BDNF protein levels in AD. PMID- 15899247 TI - Glial cell line-derived neurotrophic factor-conjugated nanoparticles suppress acquisition of cocaine self-administration in rats. AB - The neurotrophic factor glial cell line-derived neurotrophic factor (GDNF) may have therapeutic potential for preventing and treating cocaine addiction. Previously, we found that transplantation of a GDNF-expressing astrocyte cell line into the striatum and nucleus accumbens attenuates cocaine-seeking behavior in Sprague-Dawley rats. However, as a potential treatment for humans, cell transplantation presents several technical and ethical complications. Nanoparticulate systems are a safe and effective method for introducing exogenous compounds into the brain. Therefore, we examined the effect of GDNF-conjugated nanoparticles microinjected into the striatum and nucleus accumbens on cocaine self-administration in rats. GDNF-conjugated nanoparticles blocked the acquisition of cocaine self-administration compared to control treatments. Furthermore, a cocaine dose response demonstrated that decreased lever response in rats that received GDNF-conjugated nanoparticles persisted after substitution with different cocaine doses. This effect is not due to a non-specific disruption of locomotor or operant behavior, as seen following a water operant task. The current study is one of the first demonstrations that drug-conjugated nanoparticles may be effective in treating brain disorders. These findings suggest that GDNF-conjugated nanoparticles may serve as a novel potential treatment for drug addiction. PMID- 15899248 TI - Injectable intrathecal delivery system for localized administration of EGF and FGF-2 to the injured rat spinal cord. AB - The administration of growth factors (GFs) for treatment of experimental spinal cord injury (SCI) has shown limited benefits. One reason may be the mode of delivery to the injury site. We have developed a minimally invasive and safe drug delivery system (DDS) consisting of a highly concentrated collagen solution that can be injected intrathecally at the site of injury providing localized delivery of GFs. Using the injectable DDS, epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-2) were co-delivered in the subarachnoid space of Sprague-Dawley rats. The in vivo distribution of EGF and FGF-2 in both injured and uninjured animals was monitored by immunohistochemistry. Although significant differences in the distribution of EGF and FGF-2 in the spinal cord were evident, localized delivery of the GFs resulted in significantly less cavitation at the lesion epicenter and for at least 720 mum caudal to it compared to control animals without the DDS. There was also significantly more white matter sparing at the lesion epicenter in animals receiving the GFs compared to control animals. Moreover, at 14 days post-injection, delivery of the GFs resulted in significantly greater ependymal cell proliferation in the central canal immediately rostral and caudal to the lesion edge compared to controls. These results demonstrate that the injectable DDS provides a new paradigm for localized delivery of bioactive therapeutic agents to the injured spinal cord. PMID- 15899249 TI - A noninvasive ultrasonographic method to evaluate bladder function recovery in spinal cord injured rats. AB - Suprasacral spinal cord injury induces changes in the mechanical and neuronal properties of the bladder resulting in bladder areflexia followed by bladder sphincter dyssynergia and detrusor muscle hypertrophy, which lead to urinary retention and increased bladder size. These changes are most often quantified using highly skilled urodynamic techniques that involve catheterization. We investigated whether a hand-held digital ultrasound imaging system could monitor urinary retention in the bladder following spinal cord injury in adult rats. Our findings indicate that contusive spinal cord injury resulted in high residual bladder volumes that decreased and stabilized by 2 weeks post-injury but remained significantly higher than control bladder volumes up to 46 days post-injury (the longest time point examined). Post hoc analysis indicated that the degree of bladder function recovery recorded at the end of the study correlated with the degree of bladder function recovery recorded at 6 days post-injury, indicating that bladder function recovery can be predicted by analyzing bladder volume as early as 6 days post-injury. Bladder function recovery correlated with locomotor recovery as assessed using the BBB locomotor rating scale. While providing a noninvasive assessment of bladder function with no detrimental impact on locomotor function or assessment, this protocol provides researchers with a clinically relevant outcome measure for quantifying bladder function recovery after spinal cord injury or after experimental treatments for spinal cord injury. PMID- 15899250 TI - A spinal cord pathway connecting primary afferents to the segmental sympathetic outflow system. AB - The sympathetic innervation of lumbar dorsal root ganglia (DRGs) and the possible presence of spinal cord circuits connecting primary sensory afferents to the sympathetic outflow to DRGs were investigated. We used simultaneous tracing of the sympathetic input to and sensory output from DRGs. Adult male rats received unilateral microinjections of the Bartha strain of pseudorabies virus into four lumbar DRGs. At 24 h post-inoculation, productive infection was detected in both DRG neurons and sympathetic postganglionic neurons. Infection of spinal cord neurons was first observed in sympathetic preganglionic neurons of the intermediolateral column. Subsequently, the infection spread to the contralateral intermediolateral column, the area around the central canal and the superficial dorsal horn layers. To investigate the relationship between infected spinal cord neurons and primary afferents from the corresponding DRGs, we injected pseudorabies virus for retrograde tracing together with cholera toxin B for anterograde tracing. We found that infected LIV/LV and LX neurons were in close apposition to cholera toxin B labeled afferents. Importantly, immunohistochemical detection of bassoon, a pre-synaptic zone protein, identified such contacts as synapses. Together, this suggests synaptic contacts between primary sensory afferents and neurons regulating sympathetic outflow to corresponding DRGs. PMID- 15899251 TI - Vascular changes in the subventricular zone after distal cortical lesions. AB - One of the effects of cortical lesions is to produce cell proliferation in the subventricular zone (SVZ), a neurogenic zone of the adult brain distal from the lesion. The mechanisms of these effects are unknown. Recent evidence points to a relationship between the vasculature and neurogenesis both in vitro and in vivo. In the present study, we asked whether cortical lesions induced vascular modifications in the distal SVZ in vivo. Lesions of the frontoparietal cortex were produced by thermocoagulation of pial blood vessels, a method that leads to highly reproducible loss of all cortical layers, sparing the corpus callosum and underlying striatum. These lesions induced increased immunoreactivity for vascular endothelial growth factor (VEGF) around the walls of SVZ vessels, at a considerable distance from the lesion. Vascular permeability was markedly increased in both the SVZ and RMS by 3 days after the injury. A dramatic increase in endothelial proliferation was followed by expansion of the local SVZ vascular tree 7 days after the injury. This time course corresponded to the proliferative changes in the SVZ, and a tight correlation was observed between the number of blood vessels and the increase in SVZ cell number. The data demonstrate that thermocoagulatory cortical lesions induce distal vascular changes that could play a role in lesion-induced SVZ expansion. PMID- 15899252 TI - Parkinsonian action tremor: interference with object manipulation and lacking levodopa response. AB - It has been postulated that Parkinsonian action tremor is distinct from classical resting tremor and that it may contribute to a loss of manual dexterity in Parkinson's disease. We analyzed pinch grip coordination in 20 patients with Parkinson's disease. An object with and without an additional 500 g weight was grasped, lifted and held for a short time with opposed thumb and index finger. Force sensors recorded the force exerted by both fingers. Spectral analysis of the force traces was performed. Transition times between grasping and lifting the object were measured. 18 age matched normal volunteers served as a control group. While holding the object, there were force oscillations in the 3.5-6.5 Hz band indicating (reemerging) classical Parkinsonian tremor in 65% of the patients. This was reduced to 15-20% under levodopa. Oscillations in the 6-15 Hz band were found in 30% (50% with weight) of the patients, remaining unchanged under levodopa, and in 10% (20% with weight) of the normal controls. During lift initiation, 6-15 Hz oscillations were found in all patients and the majority of controls. The band power was positively correlated with the movement transition times in the severely akinetic patients and was significantly higher than in controls. It remained unchanged under levodopa. Our data confirm that Parkinsonian action tremor activated during complex voluntary movements is distinct from classical resting tremor. It does not show a clear levodopa response but affects dextrous movement coordination when associated with clinically severe overall akinesia. PMID- 15899253 TI - Quantitative assessment of forelimb motor function after cervical spinal cord injury in rats: relationship to the corticospinal tract. AB - Approximately 50% of human spinal cord injuries (SCI) are at the cervical level, resulting in impairments in motor function of the upper extremity. Even modest recovery of upper extremity function could have an enormous impact on quality of life for quadriplegics. Thus, there is a critical need to develop experimental models for cervical SCI and techniques to assess deficits and recovery of forelimb motor function. Here, we analyze forelimb and forepaw motor function in rats after a lateral hemisection at C5 and assessed the relationship between the functional impairments and the extent of damage to one descending motor system, the corticospinal tract (CST). Female Sprague-Dawley rats were trained on various behavioral tasks that require the forelimb, including a task that measures gripping ability by the hand (as measured by a grip strength meter, GSM), a food reaching task, and horizontal rope walking. After 8 weeks of post-injury testing, the distribution of the CST was evaluated by injecting BDA into the sensorimotor cortex either ipsi- or contralateral to the cervical lesion. Complete unilateral hemisection injuries eliminated the ability to grip and caused severe impairments in food retrieval by the forepaw ipsilateral to the lesion. There was no indication of recovery in either task. In cases in which hemisections spared white matter near the midline, there was some recovery of forelimb motor function over time. Assessment of rope climbing ability revealed permanent impairments in forelimb use and deficits in hindlimb use and trunk stability. Sensory testing using a dynamic plantar aesthesiometer revealed that there was no increase in touch sensitivity in the affected forelimb. For the cases in which both histological and behavioral data were available, spared forelimb motor function was greatest in rats in which there was sparing of the dorsal CST. PMID- 15899254 TI - Protective effects of melatonin against ethanol-induced reactive gliosis in hippocampus and cortex of young and aged rats. AB - Evidence has been accumulated indicating that chronic ethanol consumption leads to direct or indirect changes in the viability of central nervous system cells. The effects of aging and chronic ethanol consumption on glial markers [glial fibrillary acidic protein (GFAP) and S100B] and oxidant and antioxidant status of rats were studied. Furthermore, protective effects of melatonin against aging and alcohol consumption were also assayed. Chronic ethanol administration to young and aged rats produced an increase in lipid peroxidation, and a decline in glutathione (GSH) levels, which was significantly reversed by the co administration of melatonin. Lipid peroxidation status was markedly affected in aged rats treated with alcohol compared to the young rats. An age-related increase in GFAP and S100B levels were found in the cortex and hippocampus. Long term alcohol exposure resulted in distinct elevation in GFAP content in young rats (P < 0.01) while there was less increase in the cortex of aged rats (P < 0.05). In old rats, hippocampal GFAP levels were not significantly changed by alcohol treatment (P > 0.05). Co-administration of melatonin with alcohol significantly reduced GFAP contents both in the hippocampus (P < 0.01) and cortex (P < 0.001) of aged rats. No significant effects of alcohol treatment were found on the levels of neuron-specific enolase (NSE) in aged rats. This finding suggests that melatonin exerts its protective effect on injured nervous tissues by scavenging free radicals and stabilizing glial activity against the damaging effects of ethanol and aging. Furthermore, this work suggests that the signal to initiate gliosis is mediated, at least indirectly, by free radical formation. PMID- 15899255 TI - Pro- and anti-apoptotic evidence for cholinergic denervation and hippocampal sympathetic ingrowth in rat dorsal hippocampus. AB - In rat, injection of the specific cholinotoxin, 192 IgG-saporin, into the medial septum results not only in a selective cholinergic denervation of hippocampus, but in an ingrowth of peripheral sympathetic fibers, originating from the superior cervical ganglion, into the hippocampus. A similar process, in which peripheral noradrenergic axons invade hippocampus, may also occur in Alzheimer's disease. Since apoptotic cell death has been demonstrated in the selective neuronal loss found in Alzheimer's disease, the aim of this study was to measure apoptotic protein expression and DNA fragmentation in hippocampal sympathetic ingrowth and cholinergic denervation. Western blot, TdT-mediated dUTP nick end labeling, and oligo ligation techniques were used. Choline acetyltransferase activity and norepinephrine concentrations were also measured. As seen in our previous results, an increase in apoptotic markers was induced by cholinergic denervation alone (medial septum lesion + ganglionectomy), while hippocampal sympathetic ingrowth (medial septum + sham ganglionectomy) reduced or normalized apoptotic effects to control group levels. A decrease in choline acetyltransferase activity was also found in the dorsal hippocampus of hippocampal sympathetic ingrowth and cholinergic denervation groups. An increase in norepinephrine concentration was found in hippocampal sympathetic ingrowth but not in cholinergic denervation group. Results of this study suggest that cholinergic denervation is responsible for most of the proapoptotic responses, while hippocampal sympathetic ingrowth produces a protective effect in the process of programmed cell death in rat dorsal hippocampus. This effect may be a secondary to an altered relationship between norepinephrine-acetylcholine. PMID- 15899256 TI - The effects of osmotic stimulation and water availability on c-Fos and FosB staining in the supraoptic and paraventricular nuclei of the hypothalamus. AB - We studied the effects of osmotic stimulation on the expression of FosB and c-Fos in the supraoptic nucleus (SON) and paraventricular nucleus (PVN). Adult male rats were divided into two groups that were injected with lidocaine (0.1-0.2 ml sc) followed by either 0.9% or 6% NaCl (1 ml/100 g bw sc). After the NaCl injections, the rats were anesthetized and perfused 2, 6, or 8 h after the injections. Their brains were prepared for immunocytochemistry and stained with FosB and c-Fos antibodies. The number of c-Fos-positive cells was significantly increased only at 2 h in the SON and PVN. In contrast, the number of FosB positive cells was significantly increased at 6, and 8 h in both the SON and PVN. In a second experiment, the effect of water availability on FosB staining 8 h after injections of 6% NaCl was tested in 3 groups of rats: water ad libitum, rats that had no access to water, and rats that were given water 2 h prior to perfusion. FosB staining was significantly reduced in both the SON and the PVN of rats that had ad libitum water compared to the two water-restricted groups. In the third experiment, rats were injected with either 0.9% NaCl or 6% NaCl and were either given ad libitum access to water or water restricted for 6 h after the injections and perfused 24 h after the saline injections. FosB staining was not increased when water was available ad libitum. FosB staining was significantly increased at 24 h in the rats injected with 6% NaCl when water was restricted. Thus, FosB may continue to influence protein expression in the SON and PVN for at least 24 h following acute osmotic stimulation. PMID- 15899257 TI - Neuroprotective effect of oxidized galectin-1 in a transgenic mouse model of amyotrophic lateral sclerosis. AB - Abnormal accumulation of neurofilaments in motor neurons is a characteristic pathological finding in amyotrophic lateral sclerosis (ALS). Recently, we revealed that galectin-1, whose oxidized form has axonal regeneration-enhancing activity, accumulates in the neurofilamentous lesions in ALS. To investigate whether oxidized galectin-1 has a beneficial effect on ALS, oxidized recombinant human galectin-1 (rhGAL-1/ox) or physiological saline was injected into the left gastrocnemius muscle of the transgenic mice over-expressing a mutant copper/zinc superoxide dismutase (SOD1) with a substitution of histidine to arginine at position 46 (H46R SOD1). The H46R SOD1 transgenic mice, which represented a new animal model of familial ALS, were subsequently assessed for their disease onset, life span, duration of illness, and motor function. Furthermore, the number of remaining large anterior horn cells of spinal cords was also compared between the two groups. The results showed that administration of rhGAL-1/ox to the mice delayed the onset of their disease and prolonged the life of the mice and the duration of their illness. Motor function, as evaluated by a Rotarod performance, was improved in rhGAL-1/ox-treated mice. Significantly more anterior horn neurons of the lumbar and cervical cords were preserved in the mice injected with rhGAL 1/ox than in those injected with physiological saline. The study suggests that rhGAL-1/ox administration could be a new therapeutic strategy for ALS. PMID- 15899258 TI - The relationship between local field potential and neuronal discharge in the subthalamic nucleus of patients with Parkinson's disease. AB - Depth recordings in patients with Parkinson's disease (PD) have demonstrated prominent oscillatory activity in the beta frequency (13-35 Hz) band in local field potentials (LFPs) recorded from the region of the subthalamic nucleus (STN). Although this activity has been hypothesized to contribute to bradykinesia, it is unclear to what extent the LFP oscillations arise in the STN and are synchronous with local neuronal discharge. We therefore recorded both LFPs and multi-neuronal activity from microelectrodes inserted into STN in six PD patients (8 sides) during functional neurosurgery. As microelectrodes passed from above STN into STN, there was a pronounced increase in beta frequency band LFP activity. Furthermore, spike-triggered averages of LFP activity suggested that the discharges of neurons in STN were locked to beta oscillations in the LFP. The LFP is therefore likely to represent synchronous activity in populations of neurons in the STN of patients with PD. PMID- 15899259 TI - Electrical stimulation restores the specificity of sensory axon regeneration. AB - Electrical stimulation at the time of nerve repair promotes motoneurons to reinnervate appropriate pathways leading to muscle and stimulates sensory neurons to regenerate. The present experiments examine the effects of electrical stimulation on the specificity of sensory axon regeneration. The unoperated rat femoral cutaneous branch is served by 2-3 times more DRG neurons than is the muscle branch. After transection and repair of the femoral trunk, equal numbers of DRG neurons project to both branches. However, 1 h of electrical stimulation restores the normal proportion of DRG neurons reinnervating skin and muscle. To ask if the redistribution of stimulated neurons results from enhanced specificity of target reinnervation, we developed a new technique of sequential double labeling. DRG neurons projecting to the femoral muscle branch were prelabeled with Fluoro Gold 2 weeks before the nerve was transected proximally and repaired with or without 1 h of 20-Hz electrical stimulation. Three weeks after repair, the muscle nerve was labeled a second time with Fluororuby. The percentage of regenerating neurons that both originally served muscle and returned to muscle after nerve repair increased from 40% without stimulation to 75% with stimulation. Electrical stimulation thus dramatically alters the distribution of regenerating sensory axons, replacing normally random behavior with selective reinnervation of tissue-specific targets. If the enhanced regeneration specificity resulting from electrical stimulation is found to improve function in a large animal model, this convenient and safe technique may be a useful adjunct to clinical nerve repair. PMID- 15899260 TI - Lineage-restricted neural precursors survive, migrate, and differentiate following transplantation into the injured adult spinal cord. AB - Fetal spinal cord from embryonic day 14 (E14/FSC) has been used for numerous transplantation studies of injured spinal cord. E14/FSC consists primarily of neuronal (NRP)- and glial (GRP)-restricted precursors. Therefore, we reasoned that comparing the fate of E14/FSC with defined populations of lineage-restricted precursors will test the in vivo properties of these precursors in CNS and allow us to define the sequence of events following their grafting into the injured spinal cord. Using tissue derived from transgenic rats expressing the alkaline phosphatase (AP) marker, we found that E14/FSC exhibited early cell loss at 4 days following acute transplantation into a partial hemisection injury, but the surviving cells expanded to fill the entire injury cavity by 3 weeks. E14/FSC grafts integrated into host tissue, differentiated into neurons, astrocytes, and oligodendrocytes, and demonstrated variability in process extension and migration out of the transplant site. Under similar grafting conditions, defined NRP/GRP cells showed excellent survival, consistent migration out of the injury site and robust differentiation into mature CNS phenotypes, including many neurons. Few immature cells remained at 3 weeks in either grafts. These results suggest that by combining neuronal and glial restricted precursors, it is possible to generate a microenvironmental niche where emerging glial cells, derived from GRPs, support survival and neuronal differentiation of NRPs within the non-neurogenic and non permissive injured adult spinal cord, even when grafted into acute injury. Furthermore, the NRP/GRP grafts have practical advantages over fetal transplants, making them attractive candidates for neural cell replacement. PMID- 15899261 TI - Pleiotrophin mediates the neurotrophic effect of cyclic AMP on dopaminergic neurons: analysis of suppression-subtracted cDNA libraries and confirmation in vitro. AB - To better understand the particular vulnerability of mesencephalic dopaminergic neurons to toxins or gene mutations causing parkinsonism, we have taken advantage of a primary cell culture system in which these neurons die selectively. Antimitotic agents, such as cytosine arabinoside or cAMP, prevent the death of the neurons by arresting astrocyte proliferation. To identify factors implicated in either the death of the dopaminergic neurons or in the neuroprotective effect of cAMP, we constructed cDNA libraries enriched by subtractive hybridization and suppressive PCR in transcripts that are preferentially expressed in either control or cAMP-treated cultures. Differentially expressed transcripts were identified by hybridization of the enriched cDNAs with a commercially available cDNA expression array. The proteoglycan receptors syndecan-3 and the receptor protein tyrosine phosphatase zeta/beta were found among the transcripts preferentially expressed under control conditions, and their ligand, the cytokine pleiotrophin, was highly represented in the cDNA libraries for both conditions. Since pleiotrophin is expressed during embryonic and perinatal neural development and following lesions in the adult brain, we investigated its role in our cell culture model. Pleiotrophin was not responsible for the death of dopaminergic neurons under control conditions, or for their survival in cAMP-treated cultures. It was, however, implicated in the initial and cAMP-dependent enhancement of the differentiation of the dopaminergic neurons in our cultures. In addition, our experiments have provided evidence for a cAMP-dependent regulatory pathway leading to protease activation, and the identification of pleiotrophin as a target of this pathway. PMID- 15899262 TI - Intrathecal injection of GDNF and BDNF induces immediate early gene expression in rat spinal dorsal horn. AB - Glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) are potent trophic factors for dorsal root ganglion cells. In addition, these factors are produced in subsets of dorsal root ganglion cells and transported anterogradely to their terminals in the superficial dorsal horn of the spinal cord, where they constitute the only source of GDNF and BDNF. We investigated the effect of 10 mug GDNF and BDNF injected by lumbar puncture on the expression of the immediate early gene (IEG) products c-Fos, c-Jun, and Krox 24 in the adult rat dorsal horn. In the dorsal horn of S1 spinal segments, GDNF and BDNF induced a strong increase in IEG expression, which was most pronounced in laminae I and II (2.9- to 4.5-fold). More distal from the injection site, in the dorsal horn of L1/L2 spinal segments, the increase in IEG expression was less pronounced, suggesting a concentration-dependent effect. In order to explain the effects of intrathecally injected GDNF, we investigated whether lumbo-sacral dorsal horn neurons expressed RET protein, the signal-transducing element of the receptor complex for GDNF. It was found that several of these neurons contained RET immunoreactivity and that some of the RET-labeled neurons had the appearance of nociceptive-specific cells, confirming their presumed role in pain transmission. Additionally, using double-labeling immunofluorescence combined with confocal microscopy, it was found that after intrathecal GDNF injection 35% of c-Fos-labeled cells were also labeled for RET. These results demonstrate that intrathecally administered GDNF and BDNF induce IEG expression in dorsal horn neurons in the adult rat, supposedly by way of their cognate receptors, which are present on these neurons. We further suggest that the endogenous release of GDNF and BDNF, triggered by nociceptive stimuli, is involved in the induction of changes in spinal nociceptive transmission as in various pain states. PMID- 15899263 TI - Role of endogenous ligands for the peroxisome proliferators activated receptors alpha in the secondary damage in experimental spinal cord trauma. AB - The peroxisome proliferator-activated receptor-alpha (PPAR-alpha) is a member of the nuclear receptor superfamily of ligand-dependent transcription factors related to retinoid, steroid, and thyroid hormone receptors. The aim of the present study was to examine the effects of endogenous PPAR-alpha ligand in an experimental model of spinal cord trauma. Spinal cord injury was induced in PPAR alpha wild-type (WT) mice and PPAR-alpha knock out mice (PPAR-alpha KO) mice by the application of vascular clips (force of 24 g) to the dura via a four-level T5 T8 laminectomy. Spinal cord injury in mice resulted in severe trauma characterized by edema, neutrophil infiltration (measured as an increase in myeloperoxidase activity) and apoptosis (measured by Annexin 5 staining). An increase of immunoreactivity to TNF-alpha was observed in the spinal cord of spinal cord-injured PPAR-alpha WT mice. Absence of a functional PPAR-alpha gene in PPAR-alphaKO mice resulted in a significant augmentation of all the above described parameters. In a separate set of experiments, we have also demonstrated that the absence of PPAR-alpha gene in PPAR-alphaKO mice significantly worsened the recovery of limb function (evaluated by motor recovery score). Thus, endogenous PPAR-alpha ligands reduce the degree of development of inflammation and tissue injury events associated with spinal cord trauma in the mice. PMID- 15899264 TI - Neurotrophin-3 prevents mitochondrial dysfunction in sensory neurons of streptozotocin-diabetic rats. AB - Sensory neurons from streptozotocin (STZ)-diabetic rats exhibit depolarization of mitochondria and the related induction of reactive oxygen species has been proposed to contribute to the etiology of sensory polyneuropathy in diabetes. There is deficient neurotrophin-3 (NT-3)-dependent neurotrophic support of sensory neurons in diabetes and treatment of STZ-diabetic rats with NT-3 prevents neuropathological alterations in peripheral nerve. Therefore, we hypothesized that loss of NT-3 may contribute to mitochondrial dysfunction in sensory neurons in diabetic sensory neuropathy. The specific aim of this study was to determine whether treatment of STZ-diabetic rats with systemic NT-3 could prevent depolarization of the mitochondrial inner membrane potential (Deltapsi(m)). In vitro studies with cultured DRG neurons from control rats revealed that treatment with 50 ng/ml NT-3 for 6 h enhanced the Deltapsi(m), e.g., a higher polarized membrane potential, compared to untreated neurons (P < 0.05). Studies on DRG sensory neurons from control vs. STZ-diabetic rats demonstrated that NT-3 therapy prevented the diabetes-induced depolarization of Deltapsi(m) (P < 0.05) in parallel with normalization of diabetes-dependent deficits in sensory nerve conduction velocity. Furthermore, alterations in mitochondrial function in vitro and in vivo correlated with the level of activation/expression of Akt in DRG neurons. PMID- 15899265 TI - Differential effects of endocannabinoids on [(3)H]-GABA uptake in the rat globus pallidus. AB - In the globus pallidus, cannabinoid CB(1) receptors are localized pre synaptically on GABAergic neurons. We assessed the influence of the endocannabinoids, anandamide, 2-arachidonoyl-glycerol (2-AG) and noladin ether, on the uptake of [(3)H]-GABA in pallidal slices from rat. Both 2-AG and noladin ether increased [(3)H]-GABA uptake (by 40.8 +/- 8.0% and 38.4 +/- 12.5%). The effect of 2-AG was blocked by the cannabinoid CB(1) receptor antagonist AM 251. In contrast, neither anandamide nor the agonist WIN 55,212-2 had an effect on [(3)H]-GABA uptake. Different roles might be played by different endocannabinoids, both physiologically and in basal ganglia disorders, such as Parkinson's disease. PMID- 15899266 TI - Electricity generation using membrane and salt bridge microbial fuel cells. AB - Microbial fuel cells (MFCs) can be used to directly generate electricity from the oxidation of dissolved organic matter, but optimization of MFCs will require that we know more about the factors that can increase power output such as the type of proton exchange system which can affect the system internal resistance. Power output in a MFC containing a proton exchange membrane was compared using a pure culture (Geobacter metallireducens) or a mixed culture (wastewater inoculum). Power output with either inoculum was essentially the same, with 40+/-1mW/m2 for G. metallireducens and 38+/-1mW/m2 for the wastewater inoculum. We also examined power output in a MFC with a salt bridge instead of a membrane system. Power output by the salt bridge MFC (inoculated with G. metallireducens) was 2.2mW/m2. The low power output was directly attributed to the higher internal resistance of the salt bridge system (19920+/-50 Ohms) compared to that of the membrane system (1286+/-1Ohms) based on measurements using impedance spectroscopy. In both systems, it was observed that oxygen diffusion from the cathode chamber into the anode chamber was a factor in power generation. Nitrogen gas sparging, L-cysteine (a chemical oxygen scavenger), or suspended cells (biological oxygen scavenger) were used to limit the effects of gas diffusion into the anode chamber. Nitrogen gas sparging, for example, increased overall Coulombic efficiency (47% or 55%) compared to that obtained without gas sparging (19%). These results show that increasing power densities in MFCs will require reducing the internal resistance of the system, and that methods are needed to control the dissolved oxygen flux into the anode chamber in order to increase overall Coulombic efficiency. PMID- 15899267 TI - Use of slow filtration columns to assess oxygen respiration, consumption of dissolved organic carbon, nitrogen transformations, and microbial parameters in hyporheic sediments. AB - Biogeochemical processes mediated by microorganisms in river sediments (hyporheic sediments) play a key role in river metabolism. Because biogeochemical reactions in the hyporheic zone are often limited to the top few decimetres of sediments below the water-sediment interface, slow filtration columns were used in the present study to quantify biogeochemical processes (uptakes of O2, DOC, and nitrate) and the associated microbial compartment (biomass, respiratory activity, and hydrolytic activity) at a centimetre scale in heterogeneous (gravel and sand) sediments. The results indicated that slow filtration columns recreated properly the aerobic-anaerobic gradient classically observed in the hyporheic zone. O2 and NO3- consumptions (256 +/- 13 microg of O2 per hour and 14.6 +/- 6.1 microg of N NO3- per hour) measured in columns were in the range of values measured in different river sediments. Slow filtration columns also reproduced the high heterogeneity of the hyporheic zone with the presence of anaerobic pockets in sediments where denitrification and fermentation processes occurred. The respiratory and hydrolytic activities of bacteria were strongly linked with the O2 consumption in the experimental system, highlighting the dominance of aerobic processes in our river sediments. In comparison with these activities, the bacterial biomass (protein content) integrated both aerobic and anaerobic processes and could be used as a global microbial indicator in our system. Finally, slow filtration columns are an appropriate tool to quantify in situ rates of biogeochemical processes and to determine the relationship between the microbial compartment and the physico-chemical environment in coarse river sediments. PMID- 15899268 TI - Performance of selected anion exchange resins for the treatment of a high DOC content surface water. AB - The objective of this study was first to compare the performance of four strong anion exchange resins (AERs) (MIEX from Orica Pty Ltd, DOWEX-11 and DOWEX-MSA from DOW chemical and IRA-938 from Rohm and Haas) for their application in drinking water treatment (natural organic matter (NOM), mineral anions (nitrate, sulfate and bromide) and pesticide removal) using bench-scale experimental procedures on a high DOC content surface water. The efficiency of MIEX for NOM and mineral anions removal was furthermore evaluated using bench-scale dose response experiments on raw, clarified and post-ozonated waters. NOM removal was assessed using the measurement of dissolved organic carbon (DOC), UV absorbance at 254 nm (UV254) and the use of high-performance size exclusion chromatography with UV (HPSEC/UV) and fluorescence detection (HPSEC/FLUO). The MIEX and IRA938 anionic resins exhibit a faster removal of NOM and mineral anions compared to the DOWEX11 and MSA AERs. All the resins were found to be very effective with similar performances after 30 to 45 min of contact time. As expected, only limited sorption of atrazine and isoproturon (C0=1 microg/L) occurred with MIEX, DOWEX11 and MSA AERs. MIEX resin proved to be very efficient in eliminating NOM of high molecular weight but also a large part of the smallest UV absorbing organic compounds which were refractory to coagulation/flocculation treatment. Remaining DOC levels after 30 min of contact with MIEX were found similar in raw water, clarified water and even post-ozonated water implying no DOC benefit can be gained by employing conventional treatment prior to MIEX treatment. Removal of bromide (initial concentration 110 microg/L) was also observed and ranged from 30% to 65% for resin dose increasing from 2 to 8 mL/L. T PMID- 15899269 TI - Quantification of aqueous cyanogen chloride and cyanogen bromide in environmental samples by MIMS. AB - Membrane introduction mass spectrometry (MIMS) was developed and verified for the direct quantification of cyanogen chloride (CNCl) and cyanogen bromide (CNBr) in environmental samples without any sample workup. Factors including the membrane temperature and the liquid flow rates were examined for system optimization. The MIMS method provided linear responses for three orders of magnitude of concentrations. The instrument detection limits of CNCl and CNBr were 1.2 and 3.8 microg/L, respectively, and the method detection limits of CNCl and CNBr were both 1.7 microg/L. Effects of pH and the water matrix including synthetic water, saline water, natural surface water, and wastewater, on the responses were also examined. A pH ranging from 3 to 10 did not affect the quantification. The average recoveries of CNCl and CNBr in the water matrixes tested were 98.5% and 92.7%, respectively. The use of the MIMS method in on-line monitoring of the formation of cyanogen halide was demonstrated in chlorination of aqueous solutions containing glycine and bromide ions. The results indicated the important role of bromide ions in cyanogen halide speciation. PMID- 15899270 TI - Economical and ecological comparison of granular activated carbon (GAC) adsorber refill strategies. AB - Technical constraints can leave a considerable freedom in the design of a technology, production or service strategy. Choosing between economical or ecological decision criteria then characteristically leads to controversial solutions of ideal systems. For the adaptation of granular-activated carbon (GAC) fixed beds, various technical factors determine the adsorber volume required to achieve a desired service life. In considering carbon replacement and recycling, a variety of refill strategies are available that differ in terms of refill interval, respective adsorber volume, and time-dependent use of virgin, as well as recycled GAC. Focusing on the treatment of contaminant groundwater, we compare cost-optimal reactor configurations and refill strategies to the ecologically best alternatives. Costs and consumption of GAC are quantified within a technical economical framework. The emissions from GAC production out of hard coal, transport and recycling are equally derived through a life cycle impact assessment. It is shown how high discount rates lead to a preference of small fixed-bed volumes, and accordingly, a high number of refills. For fixed discount rates, the investigation reveals that both the economical as well as ecological assessment of refill strategies are especially sensitive to the relative valuation of virgin and recycled GAC. Since recycling results in economic and ecological benefits, optimized systems thus may differ only slightly. PMID- 15899271 TI - Oxidation and removal of arsenic (III) from aerated groundwater by filtration through sand and zero-valent iron. AB - Removing arsenic from contaminated groundwater in Bangladesh is challenging due to high concentrations of As(III), phosphate and silicate. Application of zero valent iron as a promising removal method was investigated in detail with synthetic groundwater containing 500 microg/L As(III), 2-3mg/L P, 20mg/L Si, 8.2mM HCO3-, 2.5mM Ca2+, 1.6mM Mg2+ and pH 7.0. In a series of experiments, 1L was repeatedly passed through a mixture of 1.5 g iron filings and 3-4 g quartz sand in a vertical glass column (10mm diameter), allowing the water to re-aerate between each filtration. At a flow rate of 1L/h, up to 8 mg/L dissolved Fe(II) was released. During the subsequent oxidation of Fe(II) by dissolved oxygen, As(III) was partially oxidized and As(V) sorbed on the forming hydrous ferric oxides (HFO). HFO was retained in the next filtration step and was removed by shaking of the sand-iron mixture with water. Rapid phosphate removal provided optimal conditions for the sorption of As(V). Four filtrations lead to almost complete As(III) oxidation and removal of As(tot) to below 50 microg/L. In a prototype treatment with a succession of four filters, each containing 1.5 g iron and 60 g sand, 36 L could be treated to below 50 microg/L in one continuous filtration, without an added oxidant. PMID- 15899272 TI - Activity and resistance of iron-containing amorphous, zeolitic and mesostructured materials for wet peroxide oxidation of phenol. AB - Iron-containing materials have been prepared following several strategies of synthesis and using different silica supports (amorphous, zeolitic and mesostructured materials). Activity and stability of these materials was evaluated on the wet peroxide oxidation of phenol under mild reaction conditions (100 degrees C, air pressure of 1MPa and stoichiometric amount of hydrogen peroxide for the complete mineralisation of phenol). Their catalytic performance was monitored in terms of phenol and total organic carbon (TOC) conversions, by products distribution (aromatics compounds and carboxylic acids) and degree of metal leached into the aqueous solution. The nature and local environment of iron species is strongly dependent on the synthetic route, which dramatically influences their catalytic performance. Crystalline iron oxide species supported over mesostructured SBA-15 materials have demonstrated to be the most interesting catalysts for phenol degradation according to its high organic mineralisation, low sensitivity to leaching out and good oxidant efficiency. PMID- 15899273 TI - Effects of dissolved oxygen on formation of corrosion products and concomitant oxygen and nitrate reduction in zero-valent iron systems with or without aqueous Fe2+. AB - Batch tests were conducted in zero-valent iron (ZVI or Fe0) systems to investigate oxygen consumption and the effect of dissolved oxygen (DO) on formation of iron corrosion products, nitrate reduction, the reactivity of Fe0, the role Fe2+ (aq) played, and the fate of Fe2+. The study indicates that without augmenting Fe2+ (aq), neither nitrate nor DO could be removed efficiently by Fe0. In the presence of Fe2+ (aq), nitrate and DO could be reduced concomitantly with limited interference with each other. Unlike nitrate reduction, DO removal by Fe0 did not consume Fe2+ (aq). A two-layer structure, with an inner layer of magnetite and an outer layer of lepidocrocite, may be formed in the presence of DO. When DO depleted, the outer lepidocrocite layer was transformed to magnetite. The inner layer of magnetite, even in a substantial thickness, might not impede the Fe0 reactivity as much as the thin interfacial layer between the oxide coating and liquid. Surface-bound Fe2+ may greatly enhance the electron transfer from the Fe0 core to the solid-liquid interface, and thus improve the performance of the Fe0 process. PMID- 15899274 TI - Occurrence of several acidic drugs in sewage treatment plants in Switzerland and risk assessment. AB - The occurrence and fate of five acidic drugs (Mefenamic acid, Ibuprofen, Ketoprofen, Diclofenac and Clofibric acid) were analysed in three sewage treatment plants (STP) over 4-7 consecutive days. The results point out that the five substances were persistent in wastewater effluents after municipal wastewater treatment. At the most, half of Mefenamic acid was eliminated. Ibuprofen was well removed (80%) by one sewage treatment plant. The removal of Ibuprofen is dependent on the residence time of wastewater in the STPs. A long raining period induce an important decrease of removal of Ibuprofen and Ketoprofen. Removal rates showed a great variability according to sewage treatment plants and types of treatments (e.g. biological, physico-chemical). The concentrations of Ibuprofen, Mefenamic acid and Diclofenac were relatively high in the effluents (150-2000 ng/l), showing a potential contamination of surface water. An environmental risk assessment is presented. Mefenamic acid seems to present a risk for the aquatic environment, with a ratio PEC/PNEC higher than one. PMID- 15899275 TI - Microbial partitioning to settleable particles in stormwater. AB - The degree to which microbes in the water column associate with settleable particles has important implications for microbial transport in receiving waters, as well as for microbial removal via sedimentation (i.e. detention basins). The partitioning behavior of several bacterial, protozoan and viral indicator organisms is explored in three urban streams under both storm and dry weather conditions. The fraction of organisms associated with settleable particles in stormwater is estimated through use of a centrifugation technique which is calibrated using suspensions of standard particles (e.g., glass, latex). The fraction of organisms associated with settleable particles varies by type of microbe, and the partitioning behavior of each organism generally changes between dry weather and storm conditions. Bacterial indicator organisms (fecal coliforms, Escherichia coli, enterococci) exhibited relatively consistent behavior, with an average of 20-35% of organisms associated with these particles in background samples and 30-55% in storm samples. Clostridium perfringens spores exhibited the highest average level of particle association, with storm values varying from 50% to 70%. Results related to total coliphage partitioning were more variable, with 20-60% associated with particles during storms. These estimates should be valuable in surface water quality modeling efforts, many of which currently assume that all microbes exist as free (unattached) organisms. PMID- 15899276 TI - Effect of salinity and inorganic nitrogen concentrations on nitrification and denitrification rates in intertidal sediments and rocky biofilms of the Douro River estuary, Portugal. AB - The regulatory effects of salinity and inorganic nitrogen compounds on nitrification and denitrification were studied in intertidal sandy sediments and rocky biofilms in the Douro River estuary, Portugal, over a 12-month period. Nitrification and denitrification rates were measured in slurries of field samples and enrichment experiments using the difluoromethane and the acetylene inhibition techniques, respectively. Salinity did not regulate denitrification in either environment, suggesting that halotolerant bacteria dominated the denitrifier communities. However, nitrification rates were stimulated when salinity increased from 0 to 15 practical salinity units. NO3- addition experiments revealed that NO3- availability stimulates denitrification rates in sandy sediments, but not in rocky biofilms; however, in rocky biofilms a positive and linear relationship was observed between denitrification rates and water column NO3- concentrations (r=0.92) during the monthly surveys. The N2O:N2 ratios increased rapidly when NO3- increased from 63 to 363 microM; however, results from monthly surveys showed that environmental parameters other than NO3- availability may be important in controlling the variation in N2O production via denitrification. Ammonium additions to sandy sediments stimulated nitrification rates by 35% for the 20 microM NH4+ addition, but NH4+ appeared to inhibit nitrification at high concentration addition (200 microM NH4+). In contrast, rocky biofilm nitrification was stimulated by 65% when 200 microM NH4+ was added. PMID- 15899277 TI - Blast furnace slags as sorbents of phosphate from water solutions. AB - The paper is focused on the sorption of phosphorus from aqueous solutions by crystalline and amorphous blast furnace slags. Slag sorption kinetics were measured, adsorption tests were carried out and the effect of acidification on the sorption properties of slags was studied. The kinetic measurements confirmed that the sorption of phosphorus on crystalline as well as amorphous slags can be described by a model involving pseudo-second-order reactions. For all slag types, phosphorus sorption follows the Langmuir adsorption isotherm. The acid neutralizing capacities of crystalline and amorphous slags were determined. In the case of the crystalline slags, buffering intervals were found to exist during which the slag minerals dissolve in the sequence bredigite-gehlenite-diaspor. There is a high correlation (R2=0.9989) between ANC3.8 and the saturation capacities of crystalline and amorphous slags. PMID- 15899278 TI - Removal of heavy metals from aqueous solution by nonliving Ulva seaweed as biosorbent. AB - The growth of dense green seaweed mats of Ulva spp. is an increasing problem in estuaries and coasts worldwide. The enormous amount of Ulva biomass thus becomes a troublesome waste disposal problem. On the other hand, it has been revealed that nonliving seaweed biomass, particularly brown seaweeds, has a high capacity for assimilating heavy metals. In this study, the possibility of using Ulva seaweed biomass as a biosorbent for the removal of heavy metals was examined. After processing, the biomass material was very easy to separate from the aqueous solution using a mesh. The sorption capacity of Cd on Ulva biomass increased upon pretreatment with alkali solution. The outstanding function of the biosorbent was demonstrated at around pH 8. On the basis of the Langmuir isotherms of Cd, Zn and Cu using the alkali-pretreated biomass, the parameters q(m) and b were determined to be within the narrow range of 60-90 mg/g and 0.03-0.04 L/mg, respectively, for each metal. Given the q(m) and b values, Ulva seaweed is a good biosorbent material for removing heavy metals. In an experiment using artificial wastewater containing Cd, Zn, Cu, Cr and Ni, it was possible to remove each metal simultaneously using Ulva biomass. Adsorption by Ulva biomass is effective for the removal of heavy metals from wastewater. PMID- 15899279 TI - Disinfectant efficacy of chlorite and chlorine dioxide in drinking water biofilms. AB - The drinking water industry is closely examining options to maintain disinfection in distribution systems. In particular this research compared the relative efficiency of the chlorite ion (ClO2-) to chlorine dioxide (ClO2) for biofilm control. Chlorite levels were selected for monitoring since they are typically observed in the distribution system as a by-product whenever chlorine dioxide is applied for primary or secondary disinfection. Previous research has reported the chlorite ion to be effective in mitigating nitrification in distribution systems. Annular reactors (ARs) containing polycarbonate and cast iron coupons were used to simulate water quality conditions in a distribution system. Following a 4 week acclimation period, individual ARs operated in parallel were dosed with high (0.25mg/l) and low (0.1mg/l) chlorite concentrations and with high (0.5 mg/l) and low (0.25mg/l) chlorine dioxide concentrations, as measured in the effluent of the AR. Another set of ARs that contained cast iron and polycarbonate coupons served as controls and did not receive any disinfection. The data presented herein show that the presence of chlorite at low concentration levels was not effective at reducing heterotrophic bacteria. Log reductions of attached heterotrophic bacteria for low and high chlorite ranged between 0.20 and 0.34. Chlorine dioxide had greater log reductions for attached heterotrophic bacteria ranging from 0.52 to 1.36 at the higher dose. The greatest log reduction in suspended heterotrophic bacteria was for high dose of ClO2 on either cast iron or polycarbonate coupons (1.77 and 1.55). These data indicate that it would be necessary to maintain a chlorine dioxide residual concentration in distribution systems for control of microbiological regrowth. PMID- 15899280 TI - Application of fractal flocculation and vertical transport model to aquatic sol sediment systems. AB - In estuarine and coastal environments, flocculation occurs between particles of different fractal dimensions and of different densities. Questions remain concerning the level of detail required to model particle flocculation and settling in these heterogeneous systems. This paper compares the goodness of fit between two flocculation models, using measured time series particle size distribution data collected from clay, colloidal silica, emulsified crude oil, clay-crude oil, and silica-crude oil systems. The coalesced sphere (CS) flocculation model includes the effects of heterogeneous particle size and density; the modified coalesced fractal sphere (mCFS) model adds effects due to heterogeneous fractal dimension. Goodness of fit was quantified using values of a minimized objective function, the mean of the sum of the square of the relative residuals (MSSRR). For nearly all tested experimental conditions, MSSRR values varied less than 5% between the CS and mCFS flocculation models. Additionally, collision efficiency values for single-particle-type (alpha(HOMOO)) and dual particle-type (alpha(HETT)) systems were obtained through parameter regression using the CS and mCFS models. Using the mCFS model, estimated fractal dimension (D) values obtained for clay and clay-oil systems were between 2.6 and 3.0, lower than that postulated by the CS model but higher than that estimated experimentally by the particle concentration technique. The Stokes settling velocity of a clay aggregate of a given mass is reduced with decreased fractal dimension. This results in clay-oil flocculation occurring faster than floc sedimentation in the studied hydrodynamic range. Thus, the mCFS model provides insights to the fate of spilled oil in inland and coastal waters. PMID- 15899281 TI - The residual dynamic of polycyclic aromatic hydrocarbons and organochlorine pesticides in fishponds of the Pearl River delta, South China. AB - Hong Kong and South China are the most developed regions within China. The industrialization in these areas has resulted in severe environmental problems. Sediment and biotic samples including tilapia (Oreochromis mossambicus), bighead carp (Aristichthys nobilis), grass carp (Ctenopharyngodon idellus), crucian carp (Carassius auratus) and mandarin fish/fresh water grouper (Siniperca chuatsi) were collected from different fishponds in the Pearl River Delta (Tanzhou, Sanjiao, Guangzhou, Shipai, Changan and Mai Po) for the analysis of polycyclic aromatic hydrocarbons (PAHs) and organochlorine (OC) pesticides. Mandarin fish, which belongs to the highest trophic level, accumulated the highest concentrations of PAHs and DDTs among all fish species. The levels of DDTs in fish samples ranged from 1.5 to 62ng g-1 (wet wt.), with more than 30% of the fish samples exceeding the limit of 14.4ng g-1 (wet wt.) for human consumption recommended by US EPA (2000). Levels of PAHs in fish samples ranged from 1.91 to 224.03ng g-1 (wet wt.), but the potency-weighted total concentrations of PAHs in all muscle tissues were below the guideline value of 0.67ng g-1 (wet wt.) for human consumption set by US EPA (2000). The guideline value calculated was based on a tissue consumption rate of 142.2g day-1 (4-5 meals per week), which is a more protective rate for populations with a high consumption of fish, like Chinese and Asians. The effect of lipid content in PAHs and DDTs accumulation in fish tissue was not significant in general. PMID- 15899282 TI - Determination of dichloroacetic acid and trichloroacetic acid in drinking water by acidic methanol esterification and headspace gas chromatography. AB - A simple and rapid headspace method for gas chromatographic determination of dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) in drinking water was developed. Acidic methanol esterification followed by a headspace technique using a capillary column gas chromatograph (GC) equipped with an electron capture detector (ECD) was applied to determine the levels of DCAA and TCAA in drinking water. The major advantages of this method are the use of acidic methanol as the derivatization agent instead of the hazardous diazomethane, and esterification is carried out in water instead of organic solvent. DCAA and TCAA methyl esters produced in the reaction were determined directly by a headspace GC/ECD method. The linear correlation coefficients at concentrations ranging from 0 to 60 microg/L were 0.992 and 0.996 for DCAA and TCAA, respectively. The relative standard deviations (RSD, %) for the determination of DCAA and TCAA in drinking water were 15 and 21.3%, respectively (n=3). The detection limits of this method were 3 and 0.5 microg/L for DCAA and TCAA, respectively, and the recovery was 68 103.2% for DCAA and TCAA. PMID- 15899283 TI - Adsorption of chloridazon from aqueous solution on heat and acid treated sepiolites. AB - The adsorption of chloridazon on heat treated sepiolite samples at 110 degrees C (S-110), 200 degrees C (S-200), 400 degrees C (S-400), 600 degrees C (S-600) and acid treated samples with H2SO4 solutions of two different concentrations (0.25 and 1.0M) (S-0.25 and S-1.0, respectively) from pure water at 25 degrees C has been studied by using batch experiments. In addition, column experiments were carried out with the natural (S-110) and 600 degrees C (S-600) heat treated samples, using a 10.30 mg l-1 aqueous solution of chloridazon. The adsorption experimental data points have been fitted to the Freundlich equation in order to calculate the adsorption capacities (Kf) of the samples; Kf values range from 2.89 mg kg-1 for the S-1.0 sample up to 164 mg kg-1 for the S-600 sample; so, the heat treatment given to the sepiolite greatly increases its adsorption capacity for the herbicide chloridazon whereas the acid treatment produces a clear decrease in the amount of chloridazon adsorbed. The removal efficiency (R) has also been calculated; R values ranging from 5.08% for S-1.0 up to 60.9% for S 600. The batch experiments showed that the strongest heat treatment is more effective than the natural and acid treated sepiolite in relation to adsorption of chloridazon. The column experiments also showed that 600 degrees C heat treated sepiolite might be reasonably used in removing chloridazon from water. Thus, as this type of clay is relatively plentiful, these activated samples might be reasonably used in order to remove chloridazon from water. PMID- 15899284 TI - Energy demand in sludge dewatering. AB - This work investigates the energy required to dewater a suspension, i.e., activated sludge dewatered by centrifugation or consolidation. Total energy input to the suspension from the dewatering device, bond strength between adjacent water and solid surface, and intra-cake friction loss were evaluated for original and flocculated sludges. In centrifugal dewatering, most energy input during the initial stage was consumed by overcoming process irreversibility other than intra cake friction, and, thereby, had a low energy efficiency. To increase centrifuge speed or to flocculate the sludge at optimal flocculant dosage would yield a high energy input. In the consolidation test, most energy input at the initial stage was consumed in breaking down the bond strength until the moisture content reduced to less than the critical content. During subsequent dewatering stages, friction loss became the dominant source of energy loss. Dewatering sludge with high-energy efficiency is beneficial to optimally operate a dewatering process. PMID- 15899285 TI - A novel algal toxicity testing technique for assessing the toxicity of both metallic and organic toxicants. AB - This study presents a closed-system algal toxicity test technique that is capable of detecting the effects of both organic and metallic toxicants. Toxicity testing was conducted by transferring adequate amounts of algal suspension, dilution water (with culture growth medium), and toxicants into 300-mL BOD bottles. The BOD bottles were completely filled up with no head-space left. The initial cell density and the exposure time were 15,000 cells/mL and 48 h, respectively. The performance of the above test method was evaluated using three heavy metals and six organic toxicants based on three different test endpoints, i.e., dissolved oxygen production, algal growth rate, and cell density. The proposed test revealed excellent test sensitivity and reproducibility. Currently, none of the existing algal toxicity test protocols is adequate for assessing the toxicity of organic chemicals. The closed-system algal toxicity tests developed by previous researchers also may not be ideal because the enlarged headspace and/or enriched bicarbonate buffer may result in either underestimations of the exposure concentrations or insensitive responses to both heavy metals and organic toxicants. Compared to the aforementioned algal toxicity test methods, the proposed technique in the present study has a more general applicability under conditions such as effluent samples containing both metals and organic toxicants or samples with unknown compositions. PMID- 15899286 TI - Coliform culturability in over- versus undersaturated drinking waters. AB - The culturability of Escherichia coli in undersaturated drinking water with respect to CaCO3 (corrosive water) or in oversaturated water (non-corrosive water) was tested in different reactors: glass flasks (batch, "non-reactive" wall); glass reactors (chemostat, "non-reactive" wall) versus a corroded cast iron Propella reactor (chemostat, "reactive" wall) and a 15-year-old distribution system pilot (chemostat, "reactive" wall with 1% corroded cast iron and 99% cement-lined cast iron). The E. coli in E. coli-spiked drinking water was not able to maintain its culturability and colonize the experimental systems. It appears from our results that the optimal pH for maintaining E. coli culturability was around 8.2 or higher. However, in reactors with a reactive wall (corroded cast iron), the decline in E. coli culturability was slower when the pH was adjusted to 7.9 or 7.7 (i.e. a reactor fed with corrosive water; pHpHs). We tentatively deduce that corrosion products coming from chemical reactions driven by corrosive waters on the pipe wall improve E. coli culturability. PMID- 15899287 TI - Modification of a continuous flow method for analysis of trace amounts of nitrate in iron-rich sediment pore-waters of mine pit lakes. AB - Nitrate was analysed in pore-waters with high ionic strength. Extremely high concentrations of dissolved ferrous iron interfere common analytical methods. The automatic photometrical procedure based on the cadmium reduction method is often used for analysis of nitrate in water samples (continuous flow analysis CFA). An integrated dialysis usually serves for sample dilution and (or) sample purification (from particles). Iron was precipitated as iron hydroxide due to the imidazole buffer system (pH 7.5). The dialysis membrane is an effective barrier for iron hydroxide particles to prevent interferences within the cadmium column or the flow-cell. However, dialysis membrane is blocked successively after analysis of several iron-rich pore-water samples by agglomeration of precipitated iron. The blocking of nitrate diffusion through the dialysis membrane is tantamount to a decrease of analytical sensitivity to recognise by decreasing photometrical extinction. Minimising the iron deposition within the dialyser solved the problem. A simple modification of the CFA apparatus was found to keep the analytical sensitivity nearly constant: The mixing coil for the imidazole buffer was considerably elongated. Nearly all iron hydroxide was deposited at the glass coil surface installed before the dialyser. At least 50 iron-rich samples could be analysed within one sample queue with a loss of sensitivity <10%. The recovery of nitrate was about 95%, demonstrated by spiking experiments. PMID- 15899288 TI - Role of discontinuous chlorination on microbial production by drinking water biofilms. AB - Microbial quality in water distribution systems is strongly affected by the development of microbial biofilms. Production and release of microbial cells by the biofilm affect microbial levels in the water column and in some cases this fact constitutes a public health concern. In this study, we attempt to analyze in which way the existence of different episodes of chlorine depletion affects both biofilm formation and microbial load of an artificial laboratory system. The work was carried out using two parallel packed bed reactors both supplied with running tap water. One of the reactors was used as a control and was permanently exposed to the action of chlorine. In the other reactor, chlorine was neutralized at selected times during the experiment and for periods of variable length. During the experiment the concentration of total and viable cells from the effluent was monitored at the exit of each of the reactors. The data obtained were used to estimate microbial production from the biofilms. As an average, release of microbial cells to the water phase increased tenfold in the absence of chlorine. The results also indicate that disinfectant efficiency against the biofilm was not recovered when chlorine returned to normal levels after each event of chlorine neutralization. Cell viability in the water phase in the presence of chlorine was low at the beginning of the experiment but increased 4 orders of magnitude after five neutralization periods. Therefore, subsequent episodes of chlorine depletion may accelerate the development of microbial communities with reduced susceptibility to disinfection in real drinking water systems. PMID- 15899289 TI - Removal of non-ionic organic pollutants from water via liquid-liquid extraction. AB - The removal of model pollutants bromocresol green (BG) and phenol from water is demonstrated via two liquid-liquid extraction methods. Both methods exploit selective interactions established by the pollutant molecule with a surfactant, oil, or alcohol, and are variants of the more general Winsor systems where the phases are in contact along an extremely large interfacial area. In the first method the surfactant and the co-surfactant move from a predominantly oil-in water microemulsion (Winsor I), to a middle phase microemulsion (Winsor III), and finally to a water-in-oil microemulsion (Winsor II), as the physicochemical conditions of salinity, temperature or hydrophilic-lipophilic balance of the surfactant system are varied. This method achieves better than 99% removal of the pollutant BG from water. It is argued that the removal is produced upon increasing the salinity of the system because the interaction of BG with a medium chain-length alcohol drives it to move along with the alcohol to another phase. The second method, which is scalable to industrial levels, uses a spontaneously produced water-in-oil microemulsion with large interfacial area that appears after bringing in contact water and a pre-formed Winsor II or Winsor III microemulsion system containing different surfactants and oils. The method is applied to the removal of phenol from water, and it is found that systems with polar oils such as ethyl butyrate or with cationic surfactants such as stearyl trimethylammonium chloride are more efficient in removing phenol than systems with normal alkanes or anionic surfactants. It is also shown that a microemulsion formed using a polar oil performs better than using only the polar oil as the extraction solvent. Finally, the efficiency of the second liquid-liquid extraction method can be increased from 69% in a single-stage process to 83% in a two-stage process, using the same total amount of extraction solvent. PMID- 15899290 TI - A simple model for diauxic growth of denitrifying bacteria. AB - A simple model has been formulated to simulate diauxic growth of denitrifying bacteria. It is capable of fitting the experimental results of batch growth experiments with Pseudomonas denitrificans under various conditions. It successfully predicts the observed lags when a pure culture of this bacterium switches from oxygen to nitrate as terminal electron acceptor. The model includes the effect of carbon substrate limitation and length of aerobic phase and does not run into problems when switching from anoxic to aerobic conditions, unlike prior models of diauxic growth. PMID- 15899291 TI - Mineralization of some natural refractory organic compounds by biodegradation and ozonation. AB - The objective of this study was to explore the extent of mineralization, reduction in color and reduction of COD of gallic acid, tannin and lignin by ozonation and a combination of aerobic biodegradation and ozonation. Ozonation of pure aliquots (phase I experiments) resulted in the decline in TOC, COD, COD/TOC ratio, UV absorbance at 280 nm and color of the three model compounds investigated, with COD removals of greater than 80% and high removals (>90%) of UV absorbance at 280 nm and color observed in all cases at an ozone dose of 6 mg ozone/mg initial TOC or higher. Aerobic biodegradation of pure gallic acid, tannin and lignin aliquots resulted in COD decline of approximately 36-38%. Subsequent ozonation (phase II experiments) resulted in further decline in TOC, COD, COD/TOC ratio, and increase in UV absorbance at 280 nm and color removals. COD and TOC removals comparable to phase I experiments were obtained with 30-40% lower ozone absorption in phase II experiments. The biodegradation step was quite effective in removing specific UV absorbance at 280 nm, with up to 75% removal observed. Subsequent ozonation increased overall specific UV absorbance at 280 nm to greater than 90%. PMID- 15899292 TI - Risk factors for the introduction of high pathogenicity Avian Influenza virus into poultry farms during the epidemic in the Netherlands in 2003. AB - An epidemic of high pathogenicity Avian Influenza (HPAI) occurred in the Netherlands in 2003. A census survey of 173 infected and 401 uninfected commercial poultry farms was carried out to identify factors associated with the introduction of the HPAI virus into poultry farms. Data on farm size, production characteristics, type of housing, presence of cattle and pigs were gathered by the National Inspection Service for Livestock and Meat from all farms included in this study. For each risk factor (RF) available for analysis, the Mantel-Haenszel odds ratio was calculated (stratified on farm size and housing type). We found an increased risk of HPAI virus introduction in layer finisher type poultry: OR = 2.05 (95% confidence interval, CI = 1.29-3.27). An explanation for this increased risk is the high number of contacts between these farms, especially via cardboard egg trays used for removal of eggs during the epidemic. Our analysis did not indicate significant differences between the infected and uninfected farms with regard to housing type, presence of cattle or pigs. Since layer finisher type farms are assumed to be at higher risk for HPAI virus introduction, more specific control measures might be applied in future outbreaks. PMID- 15899293 TI - Influence of tooth resection in piglets on welfare and performance. AB - In six commercial pig farms, we compared the effects of two methods of tooth resection (tooth clipping with pliers and tooth grinding with a rotating grindstone) on teeth themselves, on skin lesions of piglets and of sow udders as well as on litter growth and survival. An intact group was included for control. Treatments were balanced within herds with sows assigned to one of the three experimental treatments. Observations were from 107 sows and their litters (n = 35 or 36 litters/group) at farrowing (day 0) and approximately 8, 15 and 27 days later. Tooth resection was done within 24 h of birth after cross-fostering. Data concerning sows' lesions were analyzed on a farm basis and those concerning piglets' mortality, growth and skin lesions were analyzed on a litter basis. Frequency and severity of udder lesions differed between treatments at farrowing and on day 8; differences depended on the location of the teats (front, median or rear). Litter size and liveweight of piglets on day 0 (11.9+/-0.1 pigs, 1.51+/ 0.03 kg) and on day 27 (10.8+/-0.1 pigs, 8.08+/-0.10 kg) were similar in the three groups (mean+/-S.E.M., n = 107). Skin lesions on piglets were more frequent and/or severe in intact than in clipped piglets on days 8 and 27, whereas ground piglets had intermediate results. Because the length of the teeth was similar after clipping and grinding (P > 0.1), tooth shortening itself does not explain the differences between treatments. Overall, tooth resection had very little effect on sow mammary injuries and litter performance. It might reduce skin damage to piglets (especially, when it is performed by clipping) but teeth are severely injured. PMID- 15899294 TI - Animal and farm influences on the dynamics of antibiotic resistance in faecal Escherichia coli in young dairy calves. AB - It is believed that the intensive use of antibiotics in the management of disease in pre-weaned calves contributes to high levels of antibiotic resistance in commensal and pathogenic bacteria. We described the temporal dynamics of antibiotic-susceptibility patterns seen in bovine enteric Escherichia coli in pre weaned calves on dairy farms and dedicated calf-rearing facilities. Cohorts of 30 calves at each of six farms were sampled at 2-week intervals during the pre weaning period. Faecal E. coli isolates were analyzed for antibiotic susceptibility to 12 antibiotics with the disk-diffusion method and grouped using cluster analysis of inhibition-zone patterns. The influences of calf age, farm type, and individual-calf antibiotic therapy on the clusterings were assessed using stratified analyses and cumulative multinomial logistic regression using generalized estimating equation with antibiotic-resistance cluster as an ordinal dependent variable. The model controlled for farm and cohort by a nested design and included a repeated measure on calf at each sampling occasion. E. coli from calves 2 weeks of age were more likely to be increasingly multiply resistant than E. coli from day-old calves (OR = 53.6), as were 4- and 6-week-old calves (OR = 29.8 and 16.4, respectively). E. coli from calves on dedicated calf-rearing facilities were more likely to be increasingly multiply resistant than E. coli from dairy-reared calves (OR = 2.4). E. coli from calves treated with antibiotics within 5 days prior to sampling were also more likely to be increasingly multiply resistant than E. coli from calves not exposed to individual antibiotic therapy (OR = 2.0). PMID- 15899295 TI - Stochastic efficiency analysis of bovine tuberculosis-surveillance programs in the Netherlands. AB - We constructed a stochastic bio-economic model to determine the optimal cost efficient surveillance program for bovine tuberculosis. The surveillance programs differed in combinations of one or more detection methods and/or sampling frequency. Stochastic input variables in the epidemiological module described the dynamics of infection and the probability of detection. By means of an efficiency frontier, the trade-off between the expected cost and the epidemiological risk parameter relating to the outbreak size was evaluated. The surveillance scheme based on visual inspection of lesions on carcasses at slaughter was optimal given the current prevalence of the disease in the Netherlands if the objective was to minimise the expected costs. However, the efficient set also included two other schemes: slaughterhouse inspection in combination with GAMMA-interferon testing of blood samples and slaughterhouse inspection in combination with two-stage tuberculin testing. The choice ultimately will depend on the risk attitude of the decision-maker; a more-stringent surveillance scheme will be enforced if the expected outbreak size is to be constrained. In future scenarios, ELISA testing of bulk-tank milk in combination with the current slaughterhouse inspection procedure would outperform the surveillance scheme of solely slaughterhouse inspection if ELISA testing of bulk-tank milk becomes feasible. PMID- 15899296 TI - Spatial distribution of antibodies to Salmonella enterica serovar Typhimurium O antigens in bulk milk from Texas dairy herds. AB - Environmental factors that enhance either the survivability or dispersion of Salmonella enterica serovar Typhimurium (S. Typhimurium) could result in a spatial pattern of disease risk. The objectives of this study were to: (1) describe herd status based on antibody response to Salmonella Typhimurium as estimated from bulk tank milk samples and (2) to describe the resulting geographical patterns found among Texas dairy herds. Eight hundred and fifty-two bulk milk samples were collected from georeferenced dairy farms and assayed by an indirect enzyme-linked immunosorbent assay (ELISA) using S. Typhimurium lipopolysaccharide (LPS). ELISA signal-to-noise ratios for each bulk tank milk sample were calculated and used for geostatistical analyses. Best-fit parameters for the exponential theoretical variogram included a range of 438.8 km, partial sill of 0.060 and nugget of 0.106. The partial sill is the classical geostatistical term for the variance that can be explained by the herd's location and the nugget is the spatially random component of the variance. We have identified a spatial process in bulk milk tank titers for S. Typhimurium in Texas dairy herds and present a map of the expected smoothed surface. Areas with higher expected titers should be targeted in further studies on controlling Salmonella infection with environmental modifications. PMID- 15899297 TI - Effect of diagnostic testing error on intracluster correlation coefficient estimation. AB - Estimation of the intracluster correlation coefficient (ICC) for infectious animal diseases may be of interest for survey planning and for calculating variance inflation factors for estimators of prevalence. Typically, diagnostic tests with imperfect sensitivity and specificity are used in surveys. In such studies, where animals from multiple herds are tested, the ICC often is estimated using apparent (test-based) rather than true prevalence data. Through Monte Carlo simulation, we examined the effect of substituting diagnostic test outcomes for true infection status on an ANOVA estimator of ICC, which was designed for use with true infection status data. We considered effects of diagnostic test sensitivity and specificity on the estimated ICC when the true ICC value and infection status of the sampled individuals were known. The ANOVA estimator underestimated the true ICC when the diagnostic test was imperfect. We also demonstrated, under the beta-binomial model, that the ICC based on apparent infection status for individuals is < or = ICC based on true infection status. In addition, we propose a Bayesian model for estimating the ICC that incorporates imperfect sensitivity and specificity and illustrate the Bayesian model using a simulation study and one example; a seroprevalence survey of ovine progressive pneumonia in U.S. sheep flocks. PMID- 15899298 TI - A model of animal-human brucellosis transmission in Mongolia. AB - We developed a dynamic model of livestock-to-human brucellosis transmission in Mongolia. The compartmental model considers transmission within sheep and cattle populations and the transmission to humans as additive components. The model was fitted to demographic and seroprevalence data (Rose Bengal test) from livestock and annually reported new human brucellosis cases in Mongolia for 1991-1999 prior to the onset of a mass livestock-vaccination campaign (S19 Brucella abortus for cattle and Rev 1 Brucella melitensis for sheep and goat). The vaccination effect was fitted to livestock- and human-brucellosis data from the first 3 years of the vaccination campaign (2000-2002). Parameters were optimized on the basis of the goodness-of-fit (assessed by the deviance). The simultaneously fitted sheep-human and cattle-human contact rates show that 90% of human brucellosis was small ruminant derived. Average effective reproductive ratios for the year 1999 were 1.2 for sheep and 1.7 for cattle. PMID- 15899299 TI - An exploratory study on risk factors for postweaning multisystemic wasting syndrome (PMWS) in Spain. AB - An exploratory case-control study was carried out in Spain in 2002/2003, involving 62 pig farms of different production systems to assess risk factors that, in association with PCV2 infection, induce postweaning multisystemic wasting syndrome (PMWS) expression. To achieve this objective two groups of farms selected according to their PMWS status were compared: "cases" (farms with clinical PMWS, n = 32) and "controls" (farms without clinical PMWS, n = 30). A filled-in questionnaire and 45 blood samples (15 sows, and two groups of 15 pigs of 12 and 20 weeks of age, respectively) were obtained from each farm. Additionally, two to three diseased pigs were necropsied and relevant tissues to diagnose PMWS collected when PMWS was clinically suspected ("case" farms). A statistical analysis to compare "case" versus "control" farms was performed with the variables obtained from the questionnaire (191 variables) and the serologic test results (20 variables). Data were analysed using conditional logistic regression with a nested n:m matched design taking into account the farm size. Three variables were found significant in the final model: two related to vaccination scheme and one to PCV2 seroprevalence in growing pigs. Vaccination of gilts against PRRSV increased the odds of PMWS expression and vaccination of sows against atrophic rhinitis was related to decreased odds of the disease; however, the possibility that those two factors could be spurious effects (due to the small sample size) or confounding variables cannot be ruled out. On the other hand, a higher prevalence of antibodies to PCV2 at 12 weeks of age was observed in pigs from "case" farms than in pigs from "control" farms. This result suggests that an earlier infection with PCV2 might be a risk factor for PMWS expression. PMID- 15899300 TI - Incidence of and survival after mammary tumors in a population of over 80,000 insured female dogs in Sweden from 1995 to 2002. AB - The main objective of this study was to describe the incidence of mammary tumors (MTs) and the survival after MTs, in female dogs between 3 and 10 years of age (insured for veterinary care and with life insurance in a Swedish animal insurance company) from 1995 to 2002. Measures of incidence are presented crudely, by breed and across age categories and birth cohorts (1991-1998). The survivals until MT diagnosis and after a MT diagnosis were computed. The overall incidence for any MT claim was 111 dogs per 10,000 dog-years at risk (DYAR). The overall MT rate in the 1992 and 1993 birth cohorts was 154 dogs per 10,000 DYAR. The incidence for any MT claim increased with age and varied by breed, from 319 dogs per 10,000 DYAR in the English springer spaniel to 5 dogs per 10,000 DYAR in the rough-haired collie. At the ages 6, 8 and 10 years, 1%, 6% and 13% respectively, of all females had at least one MT claim. The MT mortality was 6 deaths per 10,000 DYAR and increased with age. The overall-case fatality was 6%. PMID- 15899301 TI - Area-level risks for BSE in British cattle before and after the July 1988 meat and bone meal feed ban. AB - In this paper we investigate area-level risk factors for BSE for the cattle population present in Great Britain between 1986 and 1997. By dividing this population into two birth cohorts, those born before the July 1988 ban on feeding ruminant-derived meat and bone meal to ruminants and those born after, second order regional influences are distinguished from the strong first-order south-to north gradient of area-level BSE risk using Bayesian hierarchical models that account for structured (spatially correlated) and unstructured heterogeneity in the data. For both cohorts area-level risk of BSE was increased by a more southerly location and greater numbers of dairy cattle, relative to non-dairy cattle. For the cohort of cattle born after the July 1988 ban on feeding ruminant derived meat and bone meal area-level BSE risk was additionally associated with greater numbers of pigs, relative to cattle. These findings support the role of low level cross-contamination of cattle feed by pig feed as an influence on BSE incidence risk as the epidemic evolved. Prior to the 1988 meat and bone meal ban unexplained BSE risk was relatively uniformly distributed across the country whereas after the ban there were spatially aggregated areas of unexplained risk in the northern and eastern regions of England suggesting that local influences allowed BSE control measures to be less-successfully applied in these areas, compared with the rest of the country. We conclude that spatially localised influences were operating in divergent ways during the two phases of the epidemic. PMID- 15899302 TI - Seroprevalence of brucellosis in mithuns (Bos frontalis) in India. AB - Studies conducted on the seroprevalence of brucellosis in 98 mithuns maintained at the National Research Centre on Mithun, Nagaland, India revealed that the number of animals found positive for brucellosis in avidin-biotin enzyme-linked immunosorbent assay, standard tube-agglutination test and Rose-Bengal plate test were 34, 20 and 11%, respectively. Highest prevalence of brucellosis was observed in the Mizoram mithun strain. The relative sensitivity and specificity of the standard tube-agglutination test were 61 and 100%, respectively; the corresponding values of the Rose-Bengal plate test were 33 and 100%, respectively. PMID- 15899303 TI - Efficacy of moxidectin injectable and pour-on formulations in a pilot control program against bovine hypodermosis in Southern Italy. AB - Bovine hypodermosis is a myiasis caused by Hypoderma bovis and Hypoderma lineatum (Diptera, Oestridae) larvae, which has a severe economic impact on the livestock industry. Though myiasis is widespread throughout Italy, no nationwide eradication program has ever been planned, unlike in other European Countries. With a view to setting up a national control program, a pilot study was carried out in Southern Italy on 9939 cattle bred in an area with a high prevalence of cattle hypodermosis, using moxidectin 0.5% pour-on (Cydectin, Fort Dodge) and 1% injectable (Cydectin, Fort Dodge) formulations. At the recommended dosage, moxidectin displayed efficacy levels of 99.9% in the pour-on and 100% in the injectable formulation, whereas the microdose (1 mg per head regardless of body weight) was less effective (65.7%). This trial contributed to a significant reduction in infestation rates in the study area and represented the first step through which a national program for eradicating warble fly infestation in Italy. PMID- 15899304 TI - Development of a simulator for radiographic image optimization. AB - A software package, incorporating two computational patient phantoms, has been developed for optimizing X-ray radiographic imaging. A tomographic phantom, visible photographic Man tomographic phantom (VIP-Man), constructed from Visible Human anatomical color images is used to simulate the scattered portion of an X ray system using the Electron Gamma Shower National Research Council (EGSnrc) Monte Carlo code. The primary portion of an X-ray image is simulated using the projection ray-tracing method through the Visible Human CT data set. To produce a realistic image, the software simulates quantum noise, blurring effects, lesions, detector absorption efficiency, and other imaging artifacts. The primary and scattered portions of an X-ray chest image are combined to form a final image for future observer studies and image quality analysis. Absorbed doses in organs and tissues of the segmented VIP-Man phantom were also obtained from the Monte Carlo simulations. This paper presents methods of the simulator and preliminary results. PMID- 15899305 TI - Sleep spindle detection using artificial neural networks trained with filtered time-domain EEG: a feasibility study. AB - An artificial neural network (ANN) based on the Multi-Layer Perceptron (MLP) architecture is used for detecting sleep spindles in band-pass filtered electroencephalograms (EEG), without feature extraction. Following optimum classification schemes, the sensitivity of the network ranges from 79.2% to 87.5%, while the false positive rate ranges from 3.8% to 15.5%. Furthermore, due to the operation of the ANN on time-domain EEG data, there is agreement with visual assessment concerning temporal resolution. Specifically, the total inter spindle interval duration and the total duration of spindles are calculated with 99% and 92% accuracy, respectively. Therefore, the present method may be suitable for investigations of the dynamics among successive inter-spindle intervals, which could provide information on the role of spindles in the sleep process, and for studies of pharmacological effects on sleep structure, as revealed by the modification of total spindle duration. PMID- 15899306 TI - Prediction of orthologous relationship by functionally important sites. AB - Making accurate functional predictions plays an important role in the era of proteomics. Reliable functional information can be extracted from orthologs in other species when annotating an unknown gene. Here a site-based approach called PORFIS is proposed to predict orthologous relationship. When applied to the bacterial transcription factor PurR/LacI family and the protein kinase AGC family, our method was able to identify, with few false positives, the important sites that agree with those verified by biological experiments. We also tested it on the alpha-proteasome family, the glycoprotein hormone family and the growth hormone family to demonstrate its ability to predict orthologous relationship. Compared with other prediction methods based on phylogenetic analysis or hidden Markov models, PORFIS not only has competitive prediction accuracy, but also provides valuable biological information of functionally important sites associated with orthologs which can be further studied in biological experiments. PMID- 15899307 TI - Yet another application of the Monte Carlo method for modeling in the field of biomedicine. AB - By means of Monte Carlo simulations performed in the C programming language, an example of scientific programming for the generation of pseudorandom numbers relevant to both teaching and research in the field of biomedicine is presented. The relatively simple algorithm proposed makes possible the statistical analysis of sequences of random numbers. The following three generators of pseudorandom numbers were used: the rand function contained in the stdlib.h library of the C programming language, Marsaglia's generator, and a chaotic function. The statistical properties of the sequences generated were compared, identical parameter values being adopted for this purpose. The properties of two estimators in finite samples of the pseudorandom numbers were also evaluated and, under suitable conditions, both the maximum-likelihood and method of moments proved to be good estimators. The findings demonstrated that the proposed algorithm appears to be suitable for the analysis of data from random experiments, indicating that it has a large variety of possible applications in the clinical practice. PMID- 15899308 TI - A program for the optimum design of pharmacokinetic, pharmacodynamic, drug metabolism and drug-drug interaction models. AB - Planning any experiment includes issues such as how many samples are to be taken and their location given some predictor variable. Often a model is used to explain these data; hence including this formally in the design will be beneficial for any subsequent parameter estimation and modelling. A number of criteria for model oriented experiments, which maximise the information content of the collected data are available. In this paper we present a program, Optdes, to investigate the optimal design of pharmacokinetic, pharmacodynamic, drug metabolism and drug-drug interaction models. Using the developed software the location of either a predetermined number of design points (exact designs) or together with the proportion of samples at each point (continuous designs) can be determined. Local as well as Bayesian designs can be optimised by either D- or A optimality criteria. Although often the optimal design cannot be applied for practical reasons, alternative designs can be readily evaluated. PMID- 15899309 TI - Analysis of esophageal atrial recordings through wavelet packets decomposition. AB - In this paper the processing of esophageal atrial electrograms by means of wavelet packets (WP) decomposition is presented. WP is described as a flexible, signal-adaptive, tool, which can be easily tuned to enhance characteristics of esophageal signals. Two aspects are mainly investigated: (i) the possibility to obtain automatic, reliable detection of atrial activation in 24h Holter recordings and (ii) the development of an algorithm for discrimination between atrial flutter (AFLU) and atrial fibrillation (AF) episodes. WP decomposition was used as a framework for pre-processing the esophageal signal and to build a set of orthonormal sub-signals which can be selected and combined according to the signal processing task to be performed: (i) in the detection of atrial activation, sub-band signal characteristics were explored at different scales by using the modulus maxima criteria and (ii) in the discrimination between AFLU and AF the coarser approximation of the esophageal signal was studied by spectral analysis. A reliable detection of atrial activation was obtained (Sensitivity (SE): 99.08%; positive predictability (+P): 98.98%). In addition a quantitative index able to discriminate between AFLU (SE: 97.5%; +P: 98.7%) and AF (SE: 98.7%; +P: 97.5%) episodes was introduced. PMID- 15899310 TI - NONMEMory: a run management tool for NONMEM. AB - NONMEM is an extremely powerful tool for nonlinear mixed-effect modelling and simulation of pharmacokinetic and pharmacodynamic data. However, it is a console based application whose output does not lend itself to rapid interpretation or efficient management. NONMEMory has been created to be a comprehensive project manager for NONMEM, providing detailed summary, comparison and overview of the runs comprising a given project, including the display of output data, simple post-run processing, fast diagnostic plots and run output management, complementary to other available modelling aids. Analysis time ought not to be spent on trivial tasks, and NONMEMory's role is to eliminate these as far as possible by increasing the efficiency of the modelling process. NONMEMory is freely available from http://www.uct.ac.za/depts/pha/nonmemory.php. PMID- 15899311 TI - 'It's caveman stuff, but that is to a certain extent how guys still operate': men's accounts of masculinity and help seeking. AB - It is often assumed that men are reluctant to seek medical care. However, despite growing interest in masculinity and men's health, few studies have focussed on men's experiences of consultation in relation to their constructions of masculinity. Those that have are largely based on men with diseases of the male body (testicular and prostate cancer) or those which have been stereotyped as male (coronary heart disease). This paper presents discussions and experiences of help seeking and its relation to, and implications for, the practice of masculinity amongst a diversity of men in Scotland, as articulated in focus group discussions. The discussions did indeed suggest a widespread endorsement of a 'hegemonic' view that men 'should' be reluctant to seek help, particularly amongst younger men. However, they also included instances which questioned or went against this apparent reluctance to seek help. These were themselves linked with masculinity: help seeking was more quickly embraced when it was perceived as a means to preserve or restore another, more valued, enactment of masculinity (e.g. working as a fire-fighter, or maintaining sexual performance or function). Few other studies have emphasised how men negotiate deviations from the hegemonic view of help-seeking. PMID- 15899312 TI - Smoking behaviour change among fathers of new infants. AB - Protecting infants from exposure to parental tobacco smoke is key to positive health outcomes in childhood and later life. While mothers' smoking has been well researched, fathers' smoking has received little attention. This paper reports data from a cross-sectional survey of 286 smoking fathers in the English Midlands, interviewed when their infants were 8-14 weeks old. It examines whether fathers attempt and successfully achieve two smoking behaviours positively associated with infant health: quitting and not smoking in the home. The birth of a new baby was not associated with attempting or successfully quitting smoking for the majority of fathers. Less than 20% had tried to quit and only 4% had successfully quit smoking since the birth of their baby. Half of the participants reported that they had not changed their cigarette consumption since their baby's birth. Not smoking in the home appeared to be a more achievable behaviour for many fathers; 78.0% had attempted and 60% had successfully achieved not smoking in home. Independent predictors of attempting to quit were fathers' own cigarette consumption and level of knowledge about infant exposure to tobacco smoke. Attempting to abstain from smoking in the home and being successful in the attempt were both independently associated with partner's smoking status, number of financially dependent children and father's social class. Findings suggest that promoting reductions in cigarette consumption and improving knowledge levels among fathers about passive smoking in infants may encourage more quit attempts. Not smoking in the home is a more achievable behaviour and is linked to fathers' caring and economic circumstances and their partner's smoking status. Influences on fathers' smoking behaviour appear to be multi-factorial. Understanding father's smoking and developing health promotion strategies to protect infants from passive smoking is likely to depend on research which can bridge the caring and economic spheres of their lives. PMID- 15899313 TI - Cultural consonance and arterial blood pressure in urban Brazil. AB - In previous research in Brazil, we tested the hypothesis that cultural consonance is associated with arterial blood pressure. Cultural consonance is the degree to which individuals are able to approximate in their own behaviors the prototypes for behavior encoded in shared cultural models. Individuals who had higher cultural consonance in the domains of lifestyle and social support had lower blood pressures. The aim of the current research was to replicate and extend these findings. First, a more extensive cultural domain analysis was carried out, improving the description of cultural models. Second, more sensitive measures of cultural consonance were developed. Third, data were collected in the same community studied previously. The following findings emerged: (a) cultural domain analysis (using a mix of quantitative and qualitative techniques) indicated that cultural models for these domains are widely shared within the community; (b) the associations of cultural consonance in these domains with arterial blood pressure were replicated; and, (c) the pattern of the associations differed slightly from that observed in earlier research. This pattern of associations can be understood in terms of macrosocial influences over the past ten years. The results support the importance of long-term fieldwork in anthropology. PMID- 15899314 TI - The impact of health care providers on female sterilization among HIV-positive women in Brazil. AB - This paper explores the reproductive preferences and outcomes of HIV-positive women in two cities in Brazil. We used three types of data, all drawn from women who delivered in public sector hospitals: (1) clinical records of 427 HIV positive women; (2) pre- and postpartum in-depth interviews with 60 HIV-positive women; and (3) a prospective survey carried out among 363 women drawn from the general population. The HIV-positive samples were collected on women who had prenatal care between July 1999 and June 2000, and the general population survey was conducted with women who started prenatal care between April 1998 and June 1999. Among the women in the clinic sample, we found dramatic differences in the proportion sterilized postpartum: 51% in Sao Paulo vs. 4% in Porto Alegre, compared to 3.4% and 1.1%, respectively, of women in the general population. Our qualitative data suggest that HIV-positive women in this study had strong preferences to have no more future children and that female sterilization was the preferred way to achieve this end. Therefore, we conclude that the large difference in rates is mainly due to HIV-positive women's differential access to sterilization in the two settings. In-depth interviews revealed that women in Sao Paulo were often encouraged by clinic staff to be sterilized postpartum. In contrast, HIV-positive women in Porto Alegre clinics were not offered sterilization as an option and those who requested it were repeatedly put off. The striking difference found in the frequency with which doctors provide postpartum sterilization to seropositive women in our study sites deserves attention and discussion in the respective medical communities. At the higher level of national policy on reproductive rights, there may be grounds for reopening discussion about the norms regarding postpartum procedures, and for devoting far more resources to expanding contraceptive options. PMID- 15899315 TI - Psychological distress among AIDS orphans in rural Uganda. AB - More than 11 million children under 15 years in sub-Saharan Africa have lost at least one parent to AIDS. In Uganda, about 2 million children are orphans, with one or both parents dead. The objective of this study was to investigate the psychosocial consequences of AIDS orphanhood in a rural district in Uganda and to identify potential areas for future interventions. The study was conducted in a randomly selected sub-county in Bushenyi District in Uganda. The study population consisted of 123 children aged 11-15 years whose parents (one or both) were reported to have died from AIDS and 110 children of similar age and gender living in intact households in the same neighbourhood. Symptoms of psychological distress were assessed using the Beck Youth Inventories of Emotional and Social Impairment (BYI). The standardized interview also included questions concerning current and past living conditions. A multivariate analysis of factors with possible relevance for BYI outcome showed that orphan status was the only significant outcome predictor. Orphans had greater risk (vs. non-orphans) for higher levels of anxiety (odds ratios (OR)=6.4), depression (OR=6.6), and anger (OR=5.1). Furthermore, orphans had significantly higher scores than non-orphans on individual items in the Beck Youth Depression Inventory that are regarded as particularly "sensitive" to the possible presence of a depressive disorder, i.e. vegetative symptoms, feelings of hopelessness, and suicidal ideation. High levels of psychological distress found in AIDS orphans suggest that material support alone is not sufficient for these children. PMID- 15899316 TI - Adherence, stereotyping and unequal HIV treatment for active users of illegal drugs. AB - Adherence to antiretroviral therapy promotes viral suppression and extends the lives of individuals with HIV, yet illegal drug users are underrepresented among eligible persons receiving HIV treatment. One explanation for this is the assumption that drug users are less capable than others of adhering to complicated medication regimens. This paper investigates this assumption by making explicit a number of its underlying propositions and examining them in light of data collected from 52 active drug users living in and around Boston, USA, who were taking highly active antiretroviral therapy for HIV (HAART). The propositions are: (1) drug users lead "chaotic" lives; (2) active drug users are always using drugs; (3) being under the influence of drugs precludes taking medications as prescribed; and (4) drug users are intrinsically different from non-users in the lives they lead and the problems they face. Data collection consisted of a series of qualitative interviews with each participant. An analytic approach informed by grounded theory was used to construct thematic content categories from the data. Results revealed stability and control as well as "chaos" in the lives of study participants. Frequency of drug use varied considerably. Using did interfere with adherence, but not in every circumstance or all cases. Not "carrying" medications, competing priorities, and re-defining regimens were the most salient of a number of non-drug-use-related obstacles observed. Documentation of patterns of variation in corresponding data highlights the stereotypical quality of the propositions. Stereotyping risks overemphasis on drug use as a barrier to adherence for active users, and underemphasis on non drug-use-related obstacles. Adherence capabilities of users, in contrast to inadequacies, are also obscured through stereotyping. As a medium for stigmatization, stereotyping may contribute to unequal treatment for drug users and other populations living with HIV. PMID- 15899317 TI - The ecology of health care in Hong Kong. AB - To better understand the distribution of resources and health care consumption patterns in different geo-ethnic and socio-economic settings, we sought to describe the patterns of illness, care-seeking behavior and health services utilization in Hong Kong compared to the US and UK. Data were derived from the 2002 Hong Kong Thematic Household Survey covering 31,762 non-institutional and institutional residents, representing 6,504,255 persons after applying population weights. Of 1000 individuals during a 1-month period, 567 reported symptoms, 512 of whom considered seeking health care. Four hundred and forty persons visited western allopathic medical practitioners, with 372 (84.5%) in primary care and 68 (15.5%) in specialty care. There were 54 visits to traditional Chinese medical practitioners and 16 emergency room episodes. Seven individuals were hospitalized in community hospitals and on average one in 1000 were admitted to a tertiary medical center. Ninety out of the 567 who experienced symptoms undertook self management strategies, which included over-the-counter western allopathic medications (n=54) or traditional Chinese remedies (n=14) or both (n=2), dietary modification (n=1) and rest (n=15). We have mapped the ecology of health care in Hong Kong. Monthly prevalence estimates were remarkably similar to US figures for hospital-based events, whereas there was evidence of apparent, substantial "over consumption" of ambulatory, community-based care. Our results also indicate that the local community's care-seeking orientation still very much favors western allopathic medicine over traditional Chinese therapy, at least for acute illness episodes. PMID- 15899318 TI - Exposure to suicide and suicidal behaviors among Hong Kong adolescents. AB - Suicidal behaviors (deliberate self-injury with the intent to hurt or kill oneself) have been little examined outside the West. The aims of this study were to (a) determine the correlates of suicidal behaviors, and (b) examine whether depression and suicide ideation moderated the effects of exposure to completed and attempted suicide on suicidal behaviors among a community sample of Hong Kong youth ages 12-17. Adolescents responded to questions regarding self-injurious behaviors, and also indicated presence of intention to hurt or kill themselves in the past 12 months. Based on their responses, two groups of interest were formed: 96 youths reported both self-injurious behaviors and the intent to hurt or kill themselves, and formed the "suicidal behaviors" group; and, 1213 adolescents reported neither self-injurious behaviors nor intent to hurt self or die, and formed the control group. The participants also responded to questions about depressive symptoms, anxiety, suicidal ideation and attempt, alcohol/drug use, stressful life events, and family relationships. They indicated whether anyone they knew had attempted or completed suicide in the previous 12 months. Logistic regression indicated that depressive symptoms, stressful life events, suicidal ideation and exposure to suicide attempt (but not completed suicide) contributed unique variance to the presence of suicidal behaviors, after controlling for demographic variables. Depression (and at trend levels, suicidal ideation) moderated the effect of exposure to suicide attempt by others on suicidal behaviors. Our results indicate that completed suicide in the social network increases risk for suicidal behaviors, but not when other risk factors are controlled. By contrast, a suicide attempt independently increases risk for suicidal behaviors. Furthermore, those youths who experience depressive symptoms or suicidal ideation are at particularly high risk for engaging in suicidal behaviors when an exposure to suicide attempt occurs. PMID- 15899319 TI - Longitudinal study of child immunization determinants in China. AB - This paper exploits longitudinal data and methods to study the determinants of child immunization in 1990 s China. Many countries such as China are experiencing rapid economic transitions characterized by declining public health expenditures, privatizing health-care sectors, increased inequality and high income growth. It is still poorly understood how such changes affect utilization of preventive health care. Data from three waves of the China Health and Nutrition Survey were used to examine the immunization effects of child, household, and community health facility characteristics, as well as changes of such effects over time. Results indicate that gender and wealth differentials in immunization increased during China's transition, though these effects were small. The most important determinants were service price and maternal education. Wealth effects were minimal, indicating that the long-run effect of economic transition on immunization rates may depend crucially on the extent to which more rapid economic growth leads to increased educational investments. Methodologically, the paper finds substantial bias from standard cross-sectional models in contrast to panel data approaches, adding to the case for expanded collection of longitudinal health data in developing countries. PMID- 15899320 TI - Measuring the quality of child health care at first-level facilities. AB - Sound policy and program decisions require timely information based on valid and relevant measures. Recent findings suggest that despite the availability of effective and affordable guidelines for the management of sick children in first level health facilities in developing countries, the quality and coverage of these services remains low. We report on the development and evaluation of a set of summary indices reflecting the quality of care received by sick children in first-level facilities. The indices were first developed through a consultative process to achieve face validity by involving technical experts and policymakers. The definition of evaluation measures for many public health programs stops at this point. We added a second phase in which standard statistical techniques were used to evaluate the content and construct validity of the indices and their reliability, drawing on data sets from the multi-country evaluation of integrated management of childhood illness (MCE) in Brazil, Tanzania and Uganda. The statistical evaluation identified important conceptual errors in the indices arising from the theory-driven expert review. The experts had combined items into inappropriate indicators resulting in summary indices that were difficult to interpret and had limited validity for program decision making. We propose a revised set of summary indices for the measurement of child health care in developing countries that is supported by both expert and statistical reviews and that led to similar programmatic insights across the three countries. We advocate increased cross-disciplinary research within public health to improve measurement approaches. Child survival policymakers, program planners and implementers can use these tools to improve their monitoring and so increase the health impact of investments in health facility care. PMID- 15899321 TI - Comparison or consumption? Distinguishing between different effects of income on health in Nordic welfare states. AB - In the relation between income and health it has been suggested that individual level mechanisms are related either to absolute or to relative income. Both absolute income level and the individual's own income in relation to that of others are likely to affect health, but to distinguish between these effects in analyses has been difficult. The aim of this study is to distinguish between the effect on health of one's own position in the income distribution and the effect on health of the individual's ability to consume. Combining data from Sweden, Finland and Norway provides a setting where individuals with the same absolute income level may occupy different positions within their national income distribution. The data come from Swedish, Finnish and Norwegian surveys of living conditions from the mid 1990s. Both the position in the income distribution and the ability to consume is measured by household disposable equivalent income. In order to eliminate differences in price levels, household income is adjusted for purchasing power parities. The outcome measure used is limiting long-standing illness. There was a clear income gradient in health over the individual's relative position in their national income distribution. Stratifying for groups of household income adjusted for purchasing power parities, we still find a significant effect of the individual's relative position. In Nordic welfare states the relative position in the income distribution is related to limiting long-standing illness independently of the ability to consume among individuals with high ability to consume. PMID- 15899322 TI - Health inequalities by wage income in Sweden: the role of work environment. AB - The main aim of this study was to explore the mediating role made by work environment to health inequalities by wage income in Sweden. Gender differences were also analysed. Data from the Swedish Survey of Living Conditions for the years 1998 and 1999 were analysed. Employed 20-64-year olds with a registered wage were included (nearly 6000 respondents). Sex-specific logistic regressions in relation to global self-rated health were applied. Those in the lowest income quintile had 2.4 times (men) and 4.3 times (women) higher probability of less than good health than did those in the highest quintile (adjusted for age, family status, country of birth, education level, smoking and full-time work). The mediating contribution of work environment factors to the health gradient by income was 25 per cent (men) and 29 per cent (women), respectively. This contribution was observed mainly from ergonomic and physical exposure, decision authority and skill discretion. Psychological demands did not contribute to such inequalities because mentally demanding work tasks are more common in high income as compared with low income jobs. Using sex-specific income quintiles, instead of income quintiles for the entire sample, gave very similar results. In conclusion, work environment factors can be seen as important mediators for the association between wage income and ill health in Sweden. A larger residual effect of income on health for women as compared with men suggests that one's own income from work is a more important determinant of women's than men's ill health in Sweden. PMID- 15899323 TI - Tobacco taxation and public health: ethical problems, policy responses. AB - This article aims to describe the major ethical issues surrounding tobacco taxation, and to identify policy responses to minimise any ethical dilemmas. It uses the standard ethical framework for biomedicine (covering beneficence, non maleficence, respect for autonomy and justice), in conjunction with relevant data on tobacco taxation from various developed countries. Tobacco taxation contributes substantial benefits at the population level by protecting health (i.e., by deterring the uptake of smoking by youth, by promoting quitting, and by reducing harm from exposure to second-hand smoke (SHS)). However, tobacco taxes can contribute to financial hardship among low-socioeconomic status populations where smoking persists. Such taxes can contribute to autonomy, by reducing SHS exposure to non-smokers, and by allowing freedom from nicotine-dependency for those who quit smoking or do not start regular smoking as a result of high tobacco prices. Furthermore, increases in tobacco taxation may reduce health inequalities and so contribute to justice. Nevertheless, the additional tax burden imposed on smokers who wish to continue to smoke, or are unable to quit, can be considered unjust. The autonomy of such smokers may be partly impaired. Although tobacco tax can be regarded as ethically justifiable because of its substantial overall benefit to society, there is substantial scope for policy changes to further reduce any harms and injustices for those populations who continue to smoke. PMID- 15899324 TI - Coding patient-centred behaviour in the medical encounter. AB - A patient-centred approach is increasingly advocated and incorporated in medical education. Due to its multi-dimensionality, however, the concept of patient centredness appears to be hard to measure and, consequently, to evaluate. The objective of this study was to develop an instrument to measure patient centredness in line with one central dimension, i.e. physicians' explorative communication skills: the tendency to encourage (or discourage) patients to express their perspective on illness and treatment, by displaying facilitating and inhibiting behaviours. The paper describes the development of the patient centred behaviour coding instrument (PBCI), and first results of validity and reliability of the instrument. The study was conducted in the outpatient division of an academic teaching hospital in The Netherlands, where follow-up encounters were videotaped and coded. Participants were 30 residents and specialists in general internal medicine, rheumatology and gastro-enterology, and 323 patients having a (video-taped) follow-up appointment with one of these physicians. All recorded consultations were coded using the PBCI. Statistical analyses verified the existence of two dimensions of the PBCI: facilitating and inhibiting behaviours. Interestingly, open and closed questions generally appeared to be indicative of both the facilitating and the inhibiting dimension; only open and closed questions with a psycho-social content were unambiguously classified as facilitating behaviours. Reliability of the facilitating behaviours was high, while reliability of the inhibiting behaviours was moderate. Besides infrequent observations of the inhibiting behaviours, low reliability was partly due to individual inter-rater variability. A global rating of patient-centredness appeared to correlate with the two dimensions in the expected direction: positively with the facilitating and negatively with the inhibiting dimension, indicating the convergent validity of the instrument. PMID- 15899325 TI - The impact of a police drug crackdown on drug injectors' ability to practice harm reduction: a qualitative study. AB - This paper employs qualitative methods to explore the ramifications of a police drug crackdown on drug injectors' ability to practice harm reduction. Between August and December 2000, we conducted open-ended interviews with 40 illicit-drug injecting residents of a New York City police precinct undergoing a crackdown. Interview topics included participants' experiences with police in the precinct and their drug use practices. Grounded theory methods were used to analyze resulting transcripts. Because place emerged as a salient analytic category, we also drew on elements of social geography to interpret results. The analysis suggests that particular crackdown tactics, notably frequent police searches of participants' bodies and elevated surveillance of the precinct's public spaces, reconfigured participants' experiences of their bodies and the public spaces comprising the precinct in ways that adversely affected their capacity to engage in harm reduction. Frequent police searches, for example, discouraged participants from carrying the injection equipment they needed to ensure that they could inject with a sterile syringe. Constant monitoring of local public spaces made it difficult for homeless women and men to inject safely. Simultaneously, participants expressed support for police actions that reduced public drug activity. Given these findings, we recommend the implementation of strategies, designed by partnerships of community groups and governmental and non governmental organizations, which reduce public drug activity without imperiling injectors' health. Possible strategies include improving access to treatment and establishing safe injection spaces. PMID- 15899326 TI - Patient preferences in randomised controlled trials: conceptual framework and implications for research. AB - Patient preferences have recently been highlighted as a potential threat to the validity of randomised controlled trials (RCTs). Although there have been significant methodological and statistical developments in relation to these issues, comparatively little attention has been paid to the development of a conceptual model concerning preferences and their effects on decision-making. This article describes the development of such a model, which was undertaken in parallel with a systematic review of the empirical data concerning preference effects. The model describes the concept of preference in terms of theoretical concepts from the psychological and economics literature, and describes a preliminary model of the development and operation of preferences in the context of RCTs. The paper then examines the implications of the model for informed consent and recruitment procedures. Key issues for future research are also outlined. PMID- 15899327 TI - Carer distress: a prospective, population-based study. AB - This study investigates whether transitions into and out of unpaid caregiving are associated with increased risk for onset of or delayed recovery from psychological distress, and traces the prevalence of distress across successive years of caring activity and after caregiving has ceased. The analysis is based on data from the British Household Panel Survey covering 3000 would-be carers, 2900 former carers, and 11,100 non-carers during the 1990s; their psychological well-being was assessed at annual intervals using the General Health Questionnaire. Carers providing long hours of care over extended spells present raised levels of distress, women more so than men. Compared with non-carers, risk for onset of distress increases progressively with the amount of time devoted to caregiving each week. Adverse effects on the psychological well-being of heavily involved carers are most pronounced around the start of their care episodes and when caregiving ends. Ongoing care increases their susceptibility to recurring distress, and adverse health effects are evident beyond the end of their caregiving episodes. Several groups of carers experience psychological health inequalities compared with non-carers, especially those looking after a spouse or partner, and mothers caring for a sick or disabled child. The findings underline the importance for effective carer support and health promotion of early identification of carers, monitoring high risk groups, timing appropriate interventions, and targeting resources. PMID- 15899328 TI - 'MMR talk' and vaccination choices: an ethnographic study in Brighton. AB - In the context of the high-profile controversy that has unfolded in the UK around the measles, mumps and rubella (MMR) vaccine and its possible adverse effects, this paper explores how parents in Brighton, southern England, are thinking about MMR for their own children. Research focusing on parents' engagement with MMR has been dominated by analysis of the proximate influences on their choices, and in particular scientific and media information, which have led health policy to focus on information and education campaigns. This paper reports ethnographic work including narratives by mothers in Brighton. Our work questions such reasoning in showing how wider personal and social issues shape parents' immunisation actions. The narratives by mothers show how practices around MMR are shaped by personal histories, by birth experiences and related feelings of control, by family health histories, by their readings of their child's health and particular strengths and vulnerabilities, by particular engagements with health services, by processes building or undermining confidence, and by friendships and conversations with others, which are themselves shaped by wider social differences and transformations. Although many see vaccination as a personal decision which must respond to the particularities of a child's immune system, 'MMR talk', which affirms these conceptualisations, has become a social phenomenon in itself. These perspectives suggest ways in which people's engagements with MMR reflect wider changes in their relations with science and the state. PMID- 15899329 TI - Structural violence and schizophrenia. AB - Despite clear evidence of a substantial biological basis to schizophrenia, there is also evidence that social, economic and political factors have considerable relevance to the clinical features, treatment and outcome of the illness. Individuals from lower socio-economic groups have an earlier age at first presentation and longer durations of untreated illness, both of which are associated with poor outcome. Individuals with schizophrenia are over-represented in the homeless population. Migration is associated with increased rates of mental illness, including schizophrenia, and this relationship appears to be mediated by psycho-social factors, including difficulties establishing social capital in smaller migrant groups. Individuals with schizophrenia are substantially over-represented amongst prison populations, and imprisonment increases the disability and stigma associated with mental illness, and impedes long-term recovery. The adverse effects of these social, economic and societal factors, along with the social stigma of mental illness, constitute a form of 'structural violence' which impairs access to psychiatric and social services and amplifies the effects of schizophrenia in the lives of sufferers. As a result of these over-arching social and economic factors, many individuals with schizophrenia are systematically excluded from full participation in civic and social life, and are constrained to live lives that are shaped by stigma, isolation, homelessness and denial of rights. There are urgent needs for (1) the development of enhanced aetiological models of schizophrenia, which elucidate the interactions between genetic risk and social environment, and can better inform bio-psycho-social approaches to treatment; (2) a renewal of emphasis on the United Nations' "Principles for the Protection of Persons with Mental Illness" and related legislative measures in individual countries; and (3) continued study and examination of the impact of social, economic and political structures on the clinical features and outcome of mental illnesses. PMID- 15899330 TI - Forecasting science futures: legitimising hope and calming fears in the embryo stem cell debate. AB - Controversies about biotechnologies often centre not so much on present scientific facts as on speculations about risks and benefits in the future. It is this key futuristic element in these arguments that is the focus of this article. We examine how competing visions of utopia or dystopia are defended through the use of diverse vocabularies, metaphors, associations and appeals to authority. Our case study explores how these rhetorical processes play out in the debate about embryo stem cell research in UK national press and TV news media. The findings show how predictions from those in favour of embryo stem cell research are supported by both hype and by anti-hype, by inconsistent appeals to the technologies' innovative status and by the selective deconstruction of concepts such as 'potential' and 'hope'. The debate also mobilises binary oppositions around reason versus emotion, science versus religion and fact versus fiction. This article highlights how traditional assertions of expertise are now combined with ideas about compassion and respect for democracy and diversity. It also highlights the fact that although news reporters are often responding to topical events the real focus is often on years, even decades ahead. Close attention to how images of the future are constructed, and the evolution of new strategies for legitimation are, we suggest, important areas of on-going research, particularly in discussions of scientific and medical developments and policy. PMID- 15899331 TI - Hypothetical influence of non-indexed Spanish journals on the impact factor of radiological journals. AB - OBJECTIVE: The aim of this study was to analyze the hypothetical changes in the 2001 impact factor of 52 radiological journals included in the Science Citation Index-Journal Citation Reports by also counting cites proceeding from 73 Spanish journals on different medical specialties. Also, to estimate the possible impact factor of the official Spanish radiology journal, Radiologia, not included in this database. MATERIALS AND METHODS: A modified 2001 impact factor of 52 radiological journals and Radiologia was obtained by adding the number of cites in 1999 and 2000 from the medical Spanish journals. Data were obtained by consulting the 2001 edition of the Journal Citation Reports in the 'Web of Science' database. RESULTS: The 16,985 bibliographical references were analysed (232 of them to radiological journals). The journal with the largest increase in its 2001 impact factor (from 1.83 to 1.90) was Radiologic Clinics of North America. European Journal of Radiology was the European journal with the highest increase (from 1.084 to 1.110) in the difference between the 2001 modified and original impact factor. The modified 2001 impact factor of the 34 American journals was statistically higher (P = 0.016) than that of the 18 European journals (1.64 versus 0.93). Differences between the 2001 modified and original impact factor were slightly higher in the American journals (no statistically significant difference). The 2001 impact factor of Radiologia was 0.056. DISCUSSION: Differences between the 2001 original and modified impact factor were small, but larger in the American journals. The 2001 impact factor of Radiologia was modest, although similar to other publications included in the Journal Citation Reports. PMID- 15899332 TI - The value of high-field MRI (3T) in the assessment of sellar lesions. AB - The aim of this study was the evaluation of the normal sellar anatomy in vitro and in vivo with high-field MRI and its application in the diagnosis of sellar pathologies in comparison to standard MRI. All high-field MR images were obtained using a 3T Bruker Medspec 30/80 Scanner with a head birdcage transmit/receive coil and an actively shielded gradient system with a maximum gradient strength of 45 mT/m. Firstly an in vitro study of the sella turcica was performed to depict normal pituitary and sellar anatomy at high field. After a pilot-study this sequence-protocol was established: A RARE sequence (TR/TE = 7790/19 ms; matrix size, 512 x 512; RARE factor = 8, FOV, 200 mm) was used for T2-weighted coronal, axial and sagittal images. A 3D gradient echo sequence with magnetization preparation (MP-RAGE, TR/TE/TI = 33.5/7.6/800 ms, matrix size, 512 x 512; FOV, 200 mm, effective slice thickness, 1.88 mm; 3 averages) was used for acquisition of T1-weighted pre- and post-contrast images. Between January 2002 and March 200458 patients were enrolled in this study. Seven patients were examined for suspected microadenoma and in 51 patients 3T MRI was used to obtain additional information about the sellar lesion already known to be present from standard MRI. In 21 cases the accuracy of the imaging findings was assessed afterwards by comparison with intraoperative findings. The infiltration of the medial cavernous sinus wall was suspected on standard MRI on 15 sides (47%), on high-field MRI on 9 sides (28%) and could be verified by intraoperative findings on 6 sides (19%). Accordingly, sensitivity to infiltration was 83% for 3T and 67% for standard MRI. Specificity was 84% for 3T and 58% for standard MRI. Moreover, high-field MRI revealed microadenomas in 7 patients with a median diameter of 4mm (range 2-9 mm). The segments of the cranial nerves were seen as mean 4 hypointense spots (range 2-5 spots) on high-field MRI in contrast to 3 spots (range 0-4 spots) on standard MRI. This difference was considerably significant (P < 0.001, Wilcoxon rank sum test). The histopathological results revealed pituitary adenoma in 16 patients and non-adenomatous sellar pathologies such as Rathke's cleft cyst, sarcoidosis, meningeoma and metastasis in 5 patients. High-field MRI is superior to standard MRI for the prediction of invasion of adjacent structures in patients with pituitary adenomas and improves surgical planning of sellar lesion. PMID- 15899333 TI - Vascular anomalies, sutures and small canals of the temporal bone on axial CT. AB - PURPOSE: Subtle bony structures, small canals and fine sutures cause sometimes problems in the analysis of CTs of the temporal bone. The aim of this study was: to analyze the visibility of subtle structures and to estimate the incidence of vascular anomalies. PATIENTS AND METHOD: We retrospectively analyzed axial scans of 223 high-resolution CTs of the temporal bone obtained as single slice or spiral CT with 1mm slice thickness. All CTs had clinical indications. Two experienced radiologists studied CTs regarding the visibility of the fine sutures, fissures and small canals and the occurrence of vascular anomalies. RESULTS: The following structures were seen commonly: sphenosquamosal suture (76%), arcuate artery canal (93%), vestibular aqueduct (89%), mastoid emissary vein (82%), singular canal (56%). Not so commonly were observed: tympanosquamosal suture (31%), mastoid canaliculus (28%), lateral sigmoid sinus (28%), petrotympanic fissure (24%), tympanomastoid suture (10%). Seldom we identified: the inferior tympanic canaliculus (6%), high jugular bulb (6%), anterior sigmoid sinus (5%), dehiscent internal carotid artery canal (2%), persistent petrosquamosal sinus (1%), dehiscent jugular bulb (1%). Persistent stapedial artery, aberrant internal carotid artery, dehiscent jugular bulb, high jugular bulb with diverticulum, anterior and dehiscent sigmoid sinus were detected in below 1% of the analyzed temporal bones. The frequency of asymmetry of the jugular foramen, which varied between 3% and 42%, depended on different criterions of size. CONCLUSION: A profound knowledge of normal anatomy and anomalies of the temporal bone avoids misinterpretation as pathological lesions and iatrogenic bleedings. PMID- 15899334 TI - Interrater reliability of sonographic examinations of orbital fractures. AB - OBJECTIVE: The purpose of this study is to determine whether there are statistically significant variations among different observers when examining fractures of the orbital walls. MATERIAL AND METHODS: From December 2003 to April 2004, 28 patients with clinically suspected orbital fractures were examined by ultrasound prospectively. The US images of the infra-orbital margins, the orbital floors, the medial and lateral orbital walls of each patient were reexamined by two independent investigators. RESULTS: Computed tomography revealed fractures of the orbital floor in 28 out of 31 patients (90.3%). The infra-orbital margins showed fractures of 14 of 31 patients (45.2%). The ultrasound examinations of the orbits by the three examiners presented satisfactory correlation regarding sensitivity and specificity. There were no significant differences between investigators. There was good agreement among the ultrasound examiners regarding the infra-orbital margins. This was not the case for the orbital floors. CONCLUSIONS: If there are clear cut clinical findings ultrasound examination could represent an alternative to computed tomography. If the clinical findings were indeterminate, computed tomography was essential as implicated by this study. Accordingly, further evaluation of ultrasound examinations of fractures of the orbital margins and floors are necessary. PMID- 15899335 TI - Emphysematous changes and normal variation in smokers and COPD patients using diffusion 3He MRI. AB - INTRODUCTION: This study aims to quantify global and regional changes of diffusive motion of 3He gas within the lung, as determined by hyperpolarized 3He MR apparent diffusion coefficient (ADC) measurement, in non-smokers, smokers and chronic obstructive pulmonary disease (COPD) patients. METHODS: Age-matched groups of six healthy non-smokers, five healthy smokers and five patients with COPD. The experiments were performed with approval from the local Research Ethics Committee. Diffusion imaging was performed following hyperpolarized 3He gas inhalation, producing ADC maps. Mean and standard deviation of the ADCs were used to compare the subject groups and assess regional variations within individuals. RESULTS: The intra-individual standard deviation of ADC in the healthy smokers was significantly larger than that of the non-smoking group (P < 0.02). Compared to the non-smoking group, COPD patients had significantly higher mean and standard deviation of ADC (P < 0.01). The mean ADC in the anterior half of the chest was systematically higher than in the posterior half in the healthy non smoking subject group. DISCUSSION: This study suggests that there are regional trends in the ADC values of healthy volunteers that may have implications for the clinical interpretation of ADC values. Less homogeneous ADC values have been detected in asymptomatic smokers, indicative of damage to the distal air spaces. PMID- 15899336 TI - The HRCT appearances of granulomatous pulmonary disease in common variable immune deficiency. AB - Approximately 10% of patients with common variable immune deficiency have systemic granulomatous disease with associated interstitial lung disease. From a population of patients with CVID attending a large tertiary referral clinic for primary immunodeficiency diseases we selected a cohort who had a restrictive defect or impaired gas transfer on pulmonary function testing and/or histologically proven granulomatous disease. HRCT scans of the thorax were reviewed retrospectively in 18 patients by two radiologists. Thirteen patients had diffuse reticulation, which varied from fine to coarse with features of fibrosis. Nodules were found in eight patients. In seven, these were associated with reticulation and in one they were an isolated finding. Bronchiectasis was found as the only abnormality in three and in addition to diffuse reticulation or nodules in another three patients. Greater appreciation of the spectrum of the radiological abnormalities in CVID patients with interstitial lung disease is important. Deteriorating lung function in patients with granulomatous CVID may be secondary to interstitial lung disease rather than bronchiectasis, and treatment should be tailored accordingly. PMID- 15899337 TI - A comparative study of 2D and 3D ultrasonography for evaluation of solid breast masses. AB - OBJECTIVE: To compare image quality and diagnostic accuracy of 2D with 3D ultrasonography in solid breast masses. METHODS AND MATERIAL: To rate image quality, two radiologists compared lesion contrast and characterization of 507 solid breast masses in 2D and 3D ultrasonography and then graded the 3D imaging in 3-point scale. To characterize the masses, the same radiologists rated the examination for clarity of margin, posterior acoustic feature, and clustered microcalcifications within a mass. In addition, the masses were assigned BI-RADS categories as proposed by the American College of Radiology, criteria using just ultrasonographic features. In the 202 pathologically confirmed cases, sensitivity, specificity, positive predictive value, negative predictive value, and false negative rate for diagnosis of breast cancer in 2D and 3D ultrasonography were assessed. Image quality and diagnostic accuracy were further evaluated according to the size of the masses. RESULTS: Two observers rated 3D imaging superior to 2D imaging in terms of lesion contrast and characterization of the masses. Especially, superiority of 3D ultrasonography in terms of image quality was increasing in more than 10 mm sized masses. However, diagnostic accuracy including sensitivity, specificity, positive predictive value, negative predictive value, and false negative rate for diagnosis of breast cancer of 3D imaging was not different from 2D imaging. CONCLUSION: In spite of superior image quality on 3D ultrasonography, it does not provide additional benefits to diagnostic accuracy for diagnosis of breast cancer. PMID- 15899338 TI - DOSIS: a Monte Carlo simulation program for dose related studies in mammography. AB - Dosimetric studies in mammography are addressed by means of a Monte Carlo simulation program. The core of this program (DOSIS: dosimetry simulation studies) is a simulation model developed using FORTRAN 90, enriched with a graphical user interface developed in MS Visual Basic. User defined mammographic technique parameters affecting breast dose are imported to the simulation model and the produced results are provided by means of both absolute (surface dose, exposure at detector plane) and relative quantities (percentage depth dose, isodose curves). The program functionality has been demonstrated in the evaluation of various mammographic examination techniques. Specifically, the influence of tube voltage and filtration on the surface dose and the exposure at detector plane has been studied utilizing a water phantom. Increase of tube voltage from 25 to 30 kVp for a Mo/Mo system resulted in a 42% decrease of the surface dose for a thick breast (6 cm), without changing the exposure at the detector plane. Use of 1.02 mm Al filter for a W anode system operating at 30 kVp resulted in a 19.1% decrease of the surface dose delivered to a 5 cm water equivalent breast. Overall, W/Al systems appear to have improved dosimetric performance, resulting up to a 65% decrease of surface dose compared to Mo/Mo systems, for identical exposures at the detector plane and breast thicknesses. PMID- 15899339 TI - Diagnosis of the depth of invasion of esophageal carcinoma using digital radiography. AB - OBJECTIVE: The purpose of our investigation was to determine the usefulness of digital radiography (DR) for diagnosing the depth of invasion of esophageal carcinoma. METHODS: We evaluated 59 patients with esophageal carcinomas who underwent DR. During continuous DR in tangential views, the most distended image of the esophagus was chosen. Percent esophageal stenosis (PES) was based on the diameter across the lesion of maximal narrowing and the average of the normal oral and anal side diameters. The maximal thickness of the tumor was measured on sequentially prepared specimens. We evaluated whether the percent of esophageal stenosis correlated with the maximal thickness of the tumor on histologic findings. Receiver-operating characteristic (ROC) curves were constructed to establish the cut-off level for PES in diagnosing the depth of tumor invasion. Accuracies for the depth of the invasion were calculated based on PES using DR. For the accuracy rate, DR was compared with endoscopy and endoscopic ultrasonography (EUS). RESULTS: There was a close correlation between PES and pathological thickness of the tumor. PES values (mean+/-S.D.) were 2.45+/-0.75% in Tis and T1a tumors, 13.3+/-10.9% in T1b tumors, 35.2+/-11.1% in T2 tumors, 55.2+/-18.1% in T3 tumors, and 86.1+/-7.5% in T4 tumors. Using the ROC analysis, 12.5, 37.5, and 44.4% were the highest cut-off values of PES for differentiating < or =T1a, < or =T1b, and < or =T2 tumors. Regarding T staging, 45 (76%) of 59 lesions were staged correctly with EUS, whereas 47 (80%) were staged correctly with DR. CONCLUSION: DR is useful for diagnosing the depth of the invasion because esophageal stenosis calculated using DR is an objective index of tumor infiltration. The accuracy rate of the depth of invasion with DR was as good as that of EUS. PMID- 15899340 TI - A comparative study to validate the use of ultrasonography and computed tomography in patients with post-operative intra-abdominal sepsis. AB - PURPOSE: To validate abdominal ultrasonography and helical computed tomography in detecting causes for sepsis in patients after abdominal surgery and to determine improved criteria for its use. MATERIALS AND METHODS: Eighty-five consecutive surgical patients primarily operated for non-infectious disease were included in this prospective study. Forty-one patients were admitted to the intensive care unit. All patients were suspected of an intra-abdominal sepsis after abdominal surgery. Both ultrasonography (US) and helical abdominal computed tomography (CT) were performed to investigate the origin of an intra-abdominal sepsis. The images of both US and CT were interpreted on a four-point scale by different radiologists or residents in radiology, the investigators were blinded of each other's test. Interpretations of US and CT were compared with a reference standard which was defined by the result of diagnostic aspiration of suspected fluid collections, (re)laparotomy, clinical course or the opinion of an independent panel. Likelihood ratios and post-test probabilities were calculated and interobserver agreement was determined using kappa statistics. RESULTS: The overall prevalence of an abdominal infection was 0.49. The likelihood ratio (LR) of a positive test-result for US was 1.33 (95% CI: 0.8-2.5) and for CT scan 2.53 (95% CI: 1.4-5.0); corresponding post-test probabilities for US 0.57 (95% CI: 0.42-0.70) and for CT 0.71 (95% CI: 0.57-0.83). The LR of a negative test-result was, respectively, 0.60 (95% CI: 0.3-1.3) and 0.18 (95% CI: 0.06-0.5); corresponding post-test probabilities for US 0.37 (95% CI: 0.20-0.57) and for CT 0.15 (95% CI: 0.06-0.32) were calculated. CONCLUSION: Computed tomography can be used as the imaging modality of choice in patients suspected of intra-abdominal sepsis after abdominal surgery. Because of the low discriminatory power ultrasonography should not be performed as initial diagnostic test. PMID- 15899341 TI - Pseudolesions of left liver lobe during helical CT examinations: prevalence and comparison between unenhanced and biphasic CT findings. AB - OBJECTIVE: Localized low attenuated areas (pseudolesions) in the medial segment of left liver lobe are not rarely seen in the screening of abdomen using helical CT. The purpose of this study was to determine the prevalence of pseudolesions in the routine helical CT of abdomen and to evaluate the morphologic and enhancement features of pseudolesions in the unenhanced and enhanced CT examinations. MATERIALS AND METHODS: We retrospectively evaluated 333 contrast enhanced abdominal CT examination of 328 patients with no known liver disease, to detect the presence of pseudolesion of liver. In the presence of unenhanced and arterial phase examinations, these images were also analyzed. The imaging criteria for pseudolesion of liver was localized low attenuated area with geometric, ovoid or nodular shaped and with no mass effect adjacent to the falciform ligament, gallbladder, or porta hepatis. Previous CT, CTAP and MR examinations were also reviewed to understand the evolution of pseudolesion in patients in whom a pseudolesion was detected in the portal phase of helical CT examination. RESULTS: We identified a pseudolesion in the 65 (19.8%) of 328 patients in portal phase of helical CT examinations. Pseudolesions were identified in the medial segment of left liver lobe adjacent to falciform ligament in the 92.8% of patients, both sides of falciform ligament in the 1.5% of patients, adjacent to porta hepatis in the 3% of patients and adjacent to gallbladder 3% of patients. These lesions had triangular shape in the 66.1% of patients, ovoid shape in the 18.6% of patients, and nodular shape in the 15.3% of patients. Unenhanced, arterial and portal phase images were exist in the 50.7% of 65 patients. The pseudolesions were not identified on the unenhanced images in the 75.7% of patients and on the arterial phase images in the 55.6% of patients. CONCLUSION: Pseudolesions around the falciform ligament are not rarely seen in the routine helical CT examination of liver and abdomen. The pseudolesions are more encountered in the portal phase of helical CT examination. These lesions seem to be likely focal fatty infiltration or perfusion defect due to venous supply variation or both. Nodular shaped pseudolesions may be interpreted as true tumors and further study may require for differential diagnosis. PMID- 15899342 TI - Doppler and gray-scale ultrasound evaluation of morphological and hemodynamic changes in liver vascualture in alcoholic patients. AB - OBJECTIVE: The aim of this study is to search sonographically for morphological and hemodynamic changes in hepatic and splanchnic vasculature of alcoholic patients having no signs of hepatic damage, and compare these with normal healthy subjects. METHODS: Thirty alcohol-dependent patients and 30 control subjects with no alcohol problem or hepatic impairment were included in the study. All patients were evaluated by gray-scale and spectral Doppler ultrasound. The diameter of the portal vein, portal venous velocity, peak systolic and end diastolic velocities of hepatic and superior mesenteric arteries were assessed. RI, PI and systolic/diastolic velocity ratios were also calculated. RESULTS: Portal vein cross-sectional area was greater in alcoholic patients compared to control group (P = 0.0012). Portal vein velocity, hepatic artery peak systolic and end diastolic velocity, superior mesenteric artery peak systolic and end diastolic velocity were significantly greater in alcoholic patients than in control group (P < or = 0.001). No statistically significant difference was detected between other parameters evaluated. CONCLUSION: In alcohol-dependent patients, some hemodynamic and morphologic changes occur in hepatic and splanchnic circulation, even before the signs of hepatic damage develop. These changes can be detected by means of Doppler and gray-scale sonogrsphy. PMID- 15899343 TI - Value of computed tomography and magnetic resonance imaging for assessing severity of liver cirrhosis secondary to viral hepatitis. AB - PURPOSE: The aim of this study was to assess the value of abdominal CT and MRI in determining the severity of cirrhosis secondary to hepatitis compared to Child Pugh classification. MATERIALS AND METHODS: The study included 23 patients who were clinically and histologically diagnosed with chronic liver disease secondary to viral hepatitis. Each patient underwent dynamic abdominal CT imaging and MRI within the same week. CT and MRI findings were retrospectively reviewed. The same parameters were used from the CT and the MR images for each patient. The parameters included liver volume index (posterior segment of the right lobe, medial and lateral segments of the left lobe), spleen volume index, ascites, portosystemic collaterals, contour irregularities of the liver and confluent fibrosis within the liver. The findings were compared with the patients' Child Pugh grades. Multiple regression analysis was used for statistical analysis. RESULTS: On MRI, liver volume index (P = 0.0001), and ascites (P = 0.009) were strongly correlated with Child-Pugh grades. With CT, only ascites was correlated with Child-Pugh grades (P = 0.002). CONCLUSION: This study indicates that liver volume index on MRI, and ascites on CT and MRI are good indicators of clinical severity of cirrhosis secondary to hepatitis. To show the effect of the other parameters, more research is needed with larger patient groups. PMID- 15899344 TI - Wet radio-frequency ablation using multiple electrodes: comparative study of bipolar versus monopolar modes in the bovine liver. AB - OBJECTIVE: To determine whether hypertonic saline (HS)-mediated bipolar radio frequency (rf) application as advantages over monopolar simultaneous and alternating rf applications for creating larger areas of coagulation necrosis. MATERIALS AND METHODS: A total of 60 rf ablations using double perfused-cooled electrodes and a 200 W generator (CC-3 model, Radionics) were performed in three different modes in explanted bovine livers: simultaneous monopolar mode (groups A and A'); alternating monopolar mode (groups B and B'); or bipolar mode (groups C and C'). Electrodes were placed at inter-electrode distances of 3 and 5 cm, and HS (6% NaCl solution) was instilled into tissue at a rate of 1 mL/min through the electrodes. rf was applied for 10 (3 cm distance) or 15 min (5 cm distance). During rf application, we measured the tissue temperature at the mid-point between the two electrodes. Dimensions of the thermal ablation zones, and temperatures were compared between the 3 groups using analysis of variance or the Kruskal-Wallis test. To compare configurations of the ablation zones in each group, the ratio of longitudinal diameter (Dl) to vertical diameter (D(v)) was calculated. RESULTS: With a 3-cm inter-electrode spacing, the D(v) between the electrodes of ablated lesions was 2.4 +/- 1.2 cm in group A, 4.5 +/- 1.0 cm in group B, and 6.1 +/- 0.9 cm in group C (P < 0.05), and at a 5-cm spacing, groups B' and C' produced a single ablation area, but group A' produced two separated ablation spheres: the D(v)s were 1.4 +/- 0.2 in group A, 2.9 +/- 1.0 mm in group B, and 6.6 +/- 0.4 cm in group C (P < 0.05). For both 5- and 3-cm spacings, the temperatures at the mid-point were higher in bipolar mode than in either monopolar simultaneous or alternating modes. The ratios of Dl/D(v) of groups A, B, and C were 2.5 +/- 0.2, 1.4 +/- 0.1, and 1.1 +/- 0.1, respectively, and the corresponding figures of groups A', B' and C' were 4.5 +/- 0.2, 2.7 +/- 0.1, and 1.1 +/- 0.1, respectively (P < 0.05). CONCLUSION: HS-enhanced bipolar rf ablation creates larger, more regular coagulation necrosis than either monopolar simultaneous or alternating rf ablation. PMID- 15899345 TI - The combined effect of radiofrequency and ethanol ablation in the management of large hepatocellular carcinoma. AB - Only a small percentage of patients with large hepatocellular carcinoma (HCC) may benefit out of surgical resection. Thus, most of these patients are in need of a form of local control, such as ethanol ablation, transarterial chemoembolization (TACE), radiofrequency thermal ablation (RF), or laser induced thermotherapy (LITT). The purpose of this study was to assess the short-term effect of sequential RF and ethanol ablation in the management of large HCC (>5 cm). Our series included 40 patients with large HCC tumors (>5 cm in diameter). We adopted a protocol of overlapping RF applications, followed by repeated ethanol ablation sessions. Our results showed that the volume of tumor coagulative necrosis initially induced by RF has significantly risen after adjuvant ethanol ablation sessions (P < 0.001). Patients who achieved complete tumor necrosis after RF ablation were 52.5% of the series. This percent has jumped to 80% of the series at the end of the protocol. This indicates that such combined protocol is more effective than RF alone. Besides, it is valuable in reducing the number of RF sessions. PMID- 15899346 TI - Percutaneous transluminal angioplasty for Brescia-Cimino hemodialysis fistula dysfunction: technical success rate, patency rate and factors that influence the results. AB - OBJECTIVE: To evaluate the initial clinical success and long-term patency rates of percutaneous transluminal angioplasty (PTA) using a venous approach for dysfunctional Brescia-Cimino fistula and to identify factors that may affect initial success and long-term patency. MATERIALS AND METHODS: A total of 99 PTA procedures were performed in retrograde fassion for 60 mature Brescia-Cimino shunts with dysfunction caused by anastomotic or peripheral outflow vein stenosis or occlusion. The initial clinical success rates were compared between stenosis and occlusion using Fisher's exact test. The Kaplan-Meier method was used to calculate the primary and secondary cumulative patency rates, and the log-rank test was used for comparison. Relative risks of patency loss according to clinical characteristics were determined with multivariate Cox models. RESULTS: The initial clinical success rate of all interventions was 92%, and the rates for stenosis and occlusion were 99 and 65%, respectively (P < 0.0001). The primary and secondary cumulative patency rates for fistulas (excluding initial failure) at 12 months were 53 and 84%, respectively. The relative risks were 5.2 (P = 0.004) for longer lesions and 4.5 (P = 0.007) for younger fistulas. The primary cumulative patency rate of four patients with a younger fistula and a longer stenosis at 4 months was 0%. CONCLUSION: Favorable primary and secondary cumulative patency rates are obtained in most patients. Long lesion length and younger age of fistulas were the two factors that reduced the patency rate after PTA. PMID- 15899347 TI - Discogenic lumbar pain: association with MR imaging and CT discography. AB - OBJECTIVE: To correlate MR and CT discography findings with pain response at provocative discography in patients with discogenic back pain. MATERIALS AND METHODS: Forty-seven patients aged 25-54 years who underwent MR imaging and subsequent CT discography (97 discs) were included in this study. MR images were retrospectively evaluated regarding disc degeneration, endplate abnormalities, facet joint osteoarthritis, and high intensity zone. During discography concordant pain was regarded as positive, whereas discordant pain and no pain were regarded as negative. MR and CT discographic findings were analyzed on the base of concordant pain using the Chi-square test. RESULTS: : Concordant pain was significantly common in the following (P < 0.05): grade 4 or 5 disc degeneration [88% (30/34) in concordant pain versus 48% (30/63) in discordant pain and no pain], high intensity zone [56% (19/34) versus 30% (19/63)], combination of above two findings [53% (18/34) versus 25% (16/63)], fissured and ruptured disc at discogram [94% (32/34) versus 57% (36/63)], and contrast beyond inner annulus at CT discogram [97% (33/34) versus 57% (36/63)]. CONCLUSION: Typical MR findings with concordant pain at discography include grade 4 or 5 disc degeneration and presence of a high intensity zone. Typical CT discography findings with concordant pain were fissured/ruptured discs and contrast extending into/beyond the outer annulus on CT. PMID- 15899348 TI - Intra-abdominal desmoplastic small round cell tumors: CT findings and clinicopathological correlations in 13 cases. AB - PURPOSE: We report computed tomography (CT) findings in 13 patients with a primary abdominal desmoplastic small round cell tumor. MATERIALS AND METHODS: 13 cases (12 men, 1 woman, mean age=24.8 years) were found in our hospital database between 1991 and 2003. Clinical, CT and histopathological features were studied retrospectively. RESULTS: Peritoneal involvement was the most common feature. In 10 cases, several lobulated peritoneal soft tissue masses (with a mean of four masses per patient) were seen. Two patients had diffused irregular peritoneal carcinomatosis without any distinct peritoneal masses. One patient had a solitary mass in the pelvic space. The main sites of peritoneal involvement were the pelvic space (n=7), omentum (n=5), retroperitoneal space (n=4), small bowel mesentery (n=3), paracolic gutter (n=2 on the right and n=1 on the left), transverse colon mesentery (n=1), peri-splenic space (n=1), peri-hepatic space (n=1). The soft tissue masses were often bulky (mean 6cm, range 1-28cm), lobulated and heterogeneous with hypodense areas (in 73% of cases). In six cases, moderate ascites was seen. In one case of pelvic involvement, unilateral hydronephrosis was seen. Adenopathies were present in seven cases at the time of the diagnosis (at intraperitoneal, retroperitoneal and pelvic sites in six patients and in the groin in one patient). Five patients had liver metastases (four lesions per case excepted one patient with 30 metastases). Associated thoracic metastases were seen in three patients. The diagnosis was confirmed with four CT-guided percutaneous biopsies. CONCLUSION: Although CT features are nonspecific, the diagnosis of desmoplastic small round cell tumor may be suspected in young men with multiple bulky heterogeneous peritoneal soft tissue masses. Imaging is useful for staging and also to guide biopsies. PMID- 15899349 TI - Sonographic evaluation of plantar fasciitis and relation to body mass index. AB - PURPOSE: We have investigated the role of sonography in the diagnosis of plantar fasciitis. MATERIALS AND METHODS: This study evaluates 39 patients with plantar fasciitis and control group of 22 healthy volunteers. The plantar fascia thickness was measured 5 mm distal to the insertion of the calcaneus of plantar aponeurosis. Qualitative parameters such as decreased echogenity, biconvexity, perifascial fluid and calcification of plantar fascia were also noted. RESULTS: Mean plantar fascia thickness was measured 2.9 mm in patients with unilateral heel pain, 2.2 mm for contralateral normal heel and 2.5mm for control group. There was a statistically significant difference between heel with plantar fasciitis, contralateral normal heel and control groups (p=0.009 and 0.0001, respectively). Mean body mass index was 28 kg/m(2) in patients with heel pain and 25 kg/m2 in control group. Body mass index measurements were significantly different between plantar fasciitis and control groups. We found reduced plantar fascia echogenity in 16 cases (41%), calcaneal spur in 20 cases (51%), biconvex appearance in two cases (5.1%) and perifascial fluid in one case (2.5%). CONCLUSION: We conclude that in patients with plantar fasciitis, ultrasound may detect relatively small differences in plantar fascia thickness even in clinically unequivocal plantar fasciitis. PMID- 15899350 TI - Diaphragmatic hernia: diagnostic approaches with review of the literature. AB - Because surgical repair is indicated for the treatment of diaphragmatic hernia (DH), preoperative imaging of the diaphragmatic defect, hernia content, and associated complications with other organ's pathologies is important. While various techniques can be used on imaging of DHs, selection of the most effective but the least invasive technique will present the most accurate findings about DH, and will facilitate the management of DH. We reviewed the diaphragmatic hernia types associated with our cases, and we discussed the preferred imaging modalities for different DHs with review of the literature. We evaluated the imaging findings of 21 DH cases. They were Morgagni's hernia (n=4), Bochdalek hernia (n=2), iatrogenic DH (n=4), traumatic DH (n=6), and hiatal hernia (n=5). Although its limited findings on DH and indirect findings about the diaphragmatic rupture, plain radiography is firstly preferred technique on DH. We found that ultrasound (US) is a useful tool on DH, on traumatic DH cases especially. Not only it shows diaphragmatic continuity and herniated organs, but also it reveals associated abdominal organ's pathologies. Computed tomography (CT) scan is most effective in many DH cases. It shows the herniated abdominal organs together with complications, such as intestinal strangulation, haemothorax, and rib fractures. We stressed that Multislice CT scan with coronal and sagittal reformatted images is the most effective and useful imaging technique on DH. With high sensitivity for soft tissue, MR imaging may be performed in the selected patients, on the late presenting DH cases or on the cases of the diagnosis still in doubt especially. PMID- 15899352 TI - Normal labor: mechanism and duration. AB - Labor is refers to the chain of physiologic events that allows a fetus to undertake its journey from the uterus to the outside world. The mean duration of a singleton preganancy is 40.0 weeks (280 days), which is dated from the first day of the last normal menstrual period. The period from 37.0 weeks (259 days) to 42.0 weeks (294 days) of gestation is regarded as "term". This article focuses on the onset progress, and mechanics of normal labor term. Topics such as preterm labor (labor before 37 weeks), postterm labor (labor after 42 weeks), and abnormal labor and delivery have not been addressed and are discussed in detail elsewhere in this issue. PMID- 15899353 TI - Labor with abnormal presentation and position. AB - Abnormal presentation and position are encountered infrequently during labor. Breech and transverse presentations should be converted to cephalic presentations by external cephalic version or delivered by cesarean section. Face, brow, and compound presentations are usually managed expectantly. Persistent occiput transverse positions are managed by rotation to anterior positions and delivered as such. Occiput posterior positions can be delivered as such or rotated to occiput anterior positions. As with any position or presentation, an obstetrician should not hesitate to abandon any rotational or operative vaginal procedure and proceed to cesarean delivery if rotation or descent does not occur with relative ease. PMID- 15899354 TI - Induction of labor. AB - The rate of labor induction continues to rise significantly in the United States because of a growing use of labor induction for postterm pregnancies and elective induction of labor. Although different types and doses of prostaglandins used for cervical ripening often initiate uterine activity, the principal role of these agents is to soften the unripe cervix independent of uterine activity. Several systematic reviews with meta-analyses have shown that prostaglandins are superior to placebo and oxytocin alone in ripening of the cervix. Numerous studies and meta-analyses have assessed misoprostol's efficacy and safety as a labor induction agent. The most appropriate dose and route of administration has not yet been confirmed. PMID- 15899355 TI - Abnormalities of the first and second stages of labor. AB - Abnormalities of the first and second stages of labor refer for the most part to abnormal progression of labor. This article discusses the risk factors, diagnoses, management options, and outcomes of the various categories of labor abnormalities, and provides an evidence-based approach where one exists. The article concentrates on the term, healthy woman carrying a singleton, vertex, normally grown fetus with no anomalies. PMID- 15899356 TI - Active management of labor. AB - A source of great controversy, the active management of labor as classically defined, is routinely misunderstood and misapplied in many clinical settings. Aggressive induction protocols, early amniotomy, operative delivery, epidural analgesia, and even early admission to labor and delivery units are actions frequently thought to be synonymous with "active management of labor". To regain an understanding of the active management of labor, one needs to examine the goal of this management scheme and become more familiar with its components. PMID- 15899357 TI - Analgesia and anesthesia in labor. AB - Whether given as an epidural, spinal, or combination, regional anesthesia is an integral part of obstetrics in the United States. A variety of drugs and dosages are used in various combinations, with no one protocol exceeding others in terms of efficacy and safety. The availability of anesthesia and analgesia has had an extraordinary impact on the field of obstetrics in the twentieth century. Knowledge of the techniques and medications used, their potential toxicities, and effects on the labor process itself can only enhance obstetricians' management of the parturient in labor. PMID- 15899358 TI - Assessment of the fetus: intermittent auscultation, electronic fetal heart rate tracing, and fetal pulse oximetry. AB - Intrapartum assessment of fetal well-being has evolved over the last 40 years, with the primary focus being fetal heart rate assessments. Despite widespread use and initial enthusiasm for the potential for heart rate monitoring to reduce fetal and neonatal mortality and morbidity, conclusive evidence of improvement in long-term outcomes is lacking. Its use is associated with an increase in operative obstetric interventions and may increase morbidity associated with such interventions. Current investigation includes fetal pulse oximetry and further analysis of specific fetal electrocardiographic changes associated with hypoxic stress. Currently it is likely that fetal heart rate monitoring will maintain its role as a common intervention in obstetric units. PMID- 15899359 TI - Intrapartum assessment of the fetus: historical and evidence-based practice. AB - The principal purpose of intrapartum fetal assessment is to detect hypoxemia ischemia, which may lead to fetal jeopardy. A quarter century ago, electronic fetal monitoring replaced intermittent auscultation as the primary method of intrapartum fetal assessment in the United States. It seems that electronic fetal monitoring will continue to be the primary mode of intrapartum fetal assessment, with intermittent auscultation used in some centers primarily for low-risk parturients. From this abridged history of intrapartum fetal monitoring, it is recognized that all surveillance methods bear inherent limitations. PMID- 15899360 TI - Cesarean delivery for nonreassuring fetal heart rate tracing. AB - An understanding of cesarean delivery for nonreassuring fetal heart rate tracing is important for several reasons. This article describes the prevalence of cesarean for nonreassuring fetal heart rate tracing and risk factors, indicates what type of fetal heart rate tracing abnormalities prompts cesarean delivery, reiterates the intrauterine resuscitation that the American College of Obstetricians and Gynecologists guidelines, and suggests steps clinicians should undertake to minimize legal liability. PMID- 15899361 TI - Chorioamnionitis and labor. AB - Chorioamnionitis is a puerperal infection that exists in histologic, clinical, and subclinical forms. This entity is believed to play a causative role in many cases of spontaneous preterm delivery. It also is associated with an increased risk of dysfunctional labor. This article focuses on the effects of chorioamnionitis on the progress of labor and the appropriate treatment of patients with clinical chorioamnionitis. The epidemiology, pathogenesis, and associated maternal and neonatal outcomes also are reviewed. PMID- 15899362 TI - Shoulder dystocia: prevention and management. AB - Knowledge of the maneuvers used for the alleviation of shoulder dystocia is relevant not only for obstetric residents and attending house staff but also for family practitioners, nurses, and nurse midwives. The performance of shoulder dystocia "drills" can be helpful not only to coordinate a teamwork approach to this obstetric emergency but also to provide an opportunity to practice the maneuvers. Shoulder dystocia continues to represent an immense area of clinical interest because it typically occurs without prediction. All patients in labor should be considered at risk for the development of shoulder dystocia. PMID- 15899363 TI - Episiotomy and vaginal trauma. AB - The era of routine episiotomy is gradually ending. Previously perceived benefits gradually have been disproved as evidence-based scientific clinical studies have shown the detrimental effects of episiotomy; however, circumstances always will exist in which prudent clinical judgment may dictate the necessity for an episiotomy. In most of these situations, however, an episiotomy often can be avoided. Perhaps more hospital perinatal review committees should evaluate episiotomy rates and strive to convince their staff to reduce their rates. We can learn to be more patient and allow the natural forces of labor to gradually stretch the perineum. In reviewing the extensive volume of published literature on episiotomy and perineal-vaginal trauma, the best advice lies in the dictum "Don't just do something, sit there!" PMID- 15899364 TI - Third stage of labor. AB - All staff involved in obstetric care should be competent in performing active management of the third stage of labor. Active management has been shown to be clearly superior to physiologic or expectant management. Comparison of the various uterotonic agents has pointed to misoprostol as a potential first-line agent in treating obstetric hemorrhage. Its cost, routes of delivery, and efficacy make it an appealing drug in urban and rural settings. By being facile with the active management of labor and the prompt recognition and treatment of postpartum hemorrhage, the morbidity from significant blood loss and subsequent surgery can be reduced significantly. PMID- 15899365 TI - Comparative sensitivities between different plasma B-type natriuretic peptide assays in patients with minimally symptomatic heart failure. AB - Plasma B-type natriuretic peptide (BNP) assays have become widely used to diagnose and manage patients with heart failure. However, differences in assay characteristics may have important implications when BNP is used as a screening test for heart failure at a specific cutoff value. We performed a prospective comparison of 2 commercially available assays--one that is a laboratory-based, microparticle enzyme immunoassay (MEIA) that uses EDTA plasma specimens and one that is a point-of-care (POC), single-use fluorescence immunoassay that uses EDTA -anticoagulated whole blood or plasma specimens-in patients with heart failure and healthy controls. Despite the overall concordance between different SNP assays for the diagnosis of heart failure, their sensitivities may differ when compared at the approved diagnostic cutoff value of 100 pg/mL. At this cutoff value, the MEIA on AxSYM demonstrated greater sensitivity than POC Triage BNP assay in minimally symptomatic patients with heart failure. Therefore, for screening purposes, cutoff values for plasma BNP or N-terminal pro-BNP levels should be specific for each assay to optimize test performance. These findings suggest that there is a relationship between the decision statistics used in screening for left ventricular dysfunction and the type of diagnostic assay used. PMID- 15899366 TI - The diagnostic utility of cardiac biomarkers in detecting myocardial infarction. AB - Cardiac biomarkers, eg, cardiac troponin, have become the standard test in combination with clinical and electrocardiographic findings for physicians to conduct prompt and effective triage of patients presenting with chest pain. Cardiac biomarkers are protein components of cell structures that are released into circulation when myocardial injury occurs. The purpose of this article is multifold. First, to identify specific cardiac biomarkers and review current guidelines based on study findings on the diagnostic utility of cardiac biomarkers in detecting myocardial infarction. Recent guidelines of the European Society of Cardiology, the American College of Cardiology, the American Heart Association, and the National Academy of Clinical Biochemistry were examined, as well as relevant studies relating to the development of these guidelines. Second, to analyze the clinical significance of cardiac biomarker measurements and the challenges with existing cardiac biomarker assays. Third, to discuss our findings regarding our evaluation of the analytical performance of a chemiluminescent microparticle immunoassay for the quantitative determination of cardiac troponin I in human serum and plasma on an automated immunoassay instrument system (ARCHITECT) to aid in the diagnosis of myocardial infarction. PMID- 15899368 TI - Understanding B-type natriuretic peptide and its role in diagnosing and monitoring congestive heart failure. AB - Congestive heart failure (CHF) is a complex clinical syndrome characterized by dysfunction of the left, right, or both ventricles, which results in the impairment of the heart's ability to circulate blood at a rate sufficient to maintain the metabolic needs of peripheral tissues and various organs. Owing to the drastic increase in cardiovascular risk factors such as obesity, diabetes, and improved survival rate after acute myocardial infarction and subsequent development of CHF in the last quarter of a century, CHF has become a major and increasing cause of death and disability in the United States. Unfortunately, the signs and symptoms are nonspecific for CHF Also, routine laboratory values, electrocardiograms, and X-rays are not always accurate enough to make the appropriate diagnosis. Recently, the US Food and Drug Administration approved a new biomarker, B-type natriuretic peptide (BNP), for the purpose of diagnosing and assessing severity of CHE BNP is synthesized, stored, and released primarily by the ventricular myocardium in response to volume expansion and pressure overload. The use of SNP, along with other diagnostic tools, can enable care providers to facilitate and optimize care of heart failure patients in a variety of clinical settings. Emerging clinical data will help further refine biomarker guided therapeutic and monitoring strategies involving BNP. PMID- 15899369 TI - Separation of modified 2'-deoxyoligonucleotides using ion-pairing reversed-phase HPLC. AB - A group of 18-mers of the same base sequence, but with differing alkyl modifications is used to investigate effects of these modifications on retention of oligonucleotides using ion-pairing reversed-phase liquid chromatography (IP RPLC). It is shown that IP-RPLC is able to distinguish between oligonucleotides differing only by a single alkyl group. The identity of the nucleobase and position and length of the alkyl adduct affect retention of the oligonucleotide. These separation phenomena result from changes in charge and hydrophobicity upon alkylation. As demonstrated in this paper; chromatographic selectivity, unique to IP-RPLC, greatly facilitates the purification process of modified oligonucleotides. PMID- 15899370 TI - Quantitative detection of ketamine, norketamine, and dehydronorketamine in urine using chemical derivatization followed by gas chromatography-mass spectrometry. AB - A repeatable and highly sensitive analytical method using gas chromatography-mass spectrometry (GC-MS) in the selected ion monitoring mode (SIM) is developed for the simultaneous detection of ketamine (KT), norketamine (NK), and newly introduced dehydronorketamine (DHNK) in urine. The test specimen along with the deuterium analogues as internal standards (IS): d4-KT for KT and d4-NK for NK/DHNK, was extracted on an automatic solid-phase extraction (SPE) apparatus. The extracted eluate then was dried and derivatized with N-methyl bis(trifluoroacetamide) (CF3CONCH3COCF3, MBTFA). Finally, the cooled derivatized solution was directly injected into the GC-MS system for analysis. The proposed process achieves high sensitivity for the detection of KT, NK, and DHNK. Correlation coefficients derived from typical calibration curves in the range of 20-2000 ng/mL are 1.000 for KT and NK, 0.999 for DHNK. The limits of detection (LODs) and limits of quantitation (LOQs) are 0.5-1.0 and 1.5-3.0, respectively. The overall method recoveries of KT, NK, and DHNK are 82.2-93.4. The intra- and inter-day run deviations are smaller than 5.0%. The analytical scheme was also applied to the determination of KT, NK, and DHNK in 20 KT suspected urine specimens, and the results reconfirm that DHNK is a main metabolite of KT. PMID- 15899371 TI - Simultaneous determination of methylephedrine and noscapine in human plasma by liquid chromatography-tandem mass spectrometry. AB - A selective and sensitive method has been developed and validated for simultaneous quantification of methylephedrine and noscapine in human plasma. Analytes were extracted from human plasma samples by liquid-liquid extraction, separated on a Diamonsil C18 column and detected by tandem mass spectrometer with an atmospheric pressure chemical ionization (APCI) interface. Diphenhydramine was used as the internal standard (I.S.). The method was found to be precise and accurate within the linear range 0.1-100 ng/ml for each analyte. The intra- and inter-day relative standard deviations (R.S.D.s) were below 5.2% for methylephedrine and 6.7% for noscapine. The inter-day relative error (RE) as determined from quality control samples (QCs) was less than 3.0% for each analyte. The assay was successfully employed in a pharmacokinetic study after an oral administration of a multicomponent formulation containing 20 mg DL methylephedrine hydrochloride, 16 mg noscapine, 300 mg paracetamol and 1mg of chlorpheniramine maleate. PMID- 15899372 TI - Determination of mycotoxins in bovine milk by liquid chromatography tandem mass spectrometry. AB - Liquid chromatographic/tandem mass spectrometric methods using pneumatically assisted electrospray ionisation (LC-ESI-MS/MS) was developed for determination of 18 mycotoxins and metabolites-ochratoxin A, zearalenone, alpha-zearalenol, beta-zearalenol, alpha-zearalanol (zeranol), beta-zearalanol (taleranol), fumonisin B1, fumonisin B2, T-2 toxin, HT-2 toxin, T-2 triol, diacetoxyscirpenol (DAS), 15-monoacetoxyscirpenol (MAS), deoxynivalenol (DON), 3 acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON), deepoxy deoxynivalenol (DOM-1) and aflatoxin M1--in milk. The mycotoxins were extracted and cleaned up simultaneously. Extraction and removal of lipophilic compounds was performed at pH 2 using a two-phase mixture of acetonitrile and hexane. The acetonitrile concentration of the aqueous phase was reduced and the pH was adjusted to 8.5 before clean up by solid phase extraction (SPE) on Oasis HLB. The toxins DON, DOM-1, 3-AcDON, 15-AcDON, ochratoxin A, zearalenone, alpha zearalenol, beta-zearalenol, alpha-zearalanol and beta-zearalanol were detected in negative ion mode after separation on a Hypersil ENV analytical column, while the toxins T-2 toxin, HT-2 toxin, T-2 triol, DAS, MAS, fumonisin B1, fumonisin B2 and aflatoxin M1 were detected in positive ion mode after separation on a Luna C18 column. Two transition products were monitored for each compound. The extraction and SPE conditions were optimised to obtain maximum recovery and minimum signal suppression/enhancement. The detection capabilities related to the transition products of lowest abundance were in the range 0.020-0.15 microg/l. The mean true recoveries were in the range 76-108% at levels of 0.2-10 microg/l. PMID- 15899373 TI - The synthesis and initial characterization of an immobilized DNA unwinding element binding (DUE-B) protein chromatographic stationary phase. AB - The DNA unwinding element binding protein (DUE-B) plays a key role in DNA replication. The DUE-B protein has been immobilized on a liquid chromatography support and the resulting immobilized protein column was used for the on-line screening of a series of steroids. The DUE-B protein was expressed with an added C-terminal sequence of six adjacent histidine residues, a His6-tag and immobilized on a chiral ligand exchange support, the CLC-L column, using Ni2+ as the coordinating metal ion. The chromatographic retentions of 12 steroids were determined on the DUE-B/CLC-L column. The magnitudes of the steroid-immobilized DUE-B interactions, reflected by the observed retention times, correlated to the effect of the steroids in the cell-free replication system, i.e. the longer the retention, the greater the increase in DNA replication. The coefficient of determination for the %DNA activities linear relation to retention time was 0.9694. The data suggest that the DUE-B/CLC-L phase can be used for on-line pharmacological studies. The results also indicated that His-tagged proteins can be directly immobilized on the CLC-L stationary phase and the resulting columns used as rapid screens for the isolation and identification of small molecule or protein ligands from complex biological or chemical mixtures. PMID- 15899374 TI - One-step purification of metallothionein extracted from two different sources. AB - We describe a one-step purification of hepatic metallothionein from the Amazon fish Colossoma macropomum injected with cadmium and from the copper-loaded metallothionein from the yeast Saccharomyces cerevisiae, performed by affinity chromatography through metal-chelating columns. Yeast metallothionein was purified from Cu2+-loaded resin and eluted by a continuous EDTA gradient whereas hepatic metallothionein extracted from fishes was purified by Ni2+-loaded resin and eluted by a continuous imidazol gradient. Purified metallothioneins were evaluated by SDS-PAGE and characterized by UV spectra of the apo- and Cd2+-loaded protein. This method allowed high purity and yield as well as rapid one-step extraction of both metal-loaded and apoprotein. PMID- 15899375 TI - A sensitive and selective LC-MS-MS method for simultaneous determination of picroside-I and kutkoside (active principles of herbal preparation picroliv) using solid phase extraction in rabbit plasma: application to pharmacokinetic study. AB - A rapid, sensitive and selective LC-MS-MS method for the simultaneous quantitation of picroside-I and kutkoside (active constituents of herbal hepatoprotectant picroliv) was developed and validated in rabbit plasma. The analytes and internal standard (Amarogentin) were extracted using Oasis HLB solid phase extraction cartridges. Analysis was performed on Spheri RP-18 column (10 microm, 100 mm x 4.6 mm i.d.) coupled with guard column using acetonitrile:MilliQ water (50:50, %v/v) as mobile phase at a flow rate of 1 ml/min with a retention time of 1.39, 1.33 and 1.42 min for picroside-I, kutkoside and amarogentin, respectively. The quantitation was carried out using an API-4000 LC-MS-MS with negative electro spray ionization in multiple reaction monitoring (MRM) mode. The precursor to product ion transitions for picroside-I, kutkoside and amarogentin were m/z 491 > 147, 199; 511 > 167, 235; 585 > 227, respectively. The method was validated in terms of establishing linearity, specificity, sensitivity, recovery, accuracy and precision (within- and between-assay variation), freeze-thaw (f-t), auto injector and dry residue stability. Linearity in plasma was observed over a concentration range of 1.56-400 ng/ml with a limit of detection (LOD) of 0.5 ng/ml for both analytes. The recoveries from spiked control samples were > 60 and > 70% for picroside-I and kutkoside, respectively. Accuracy and precision of the validated method were within the acceptable limits of < 20% at low and < 15% at other concentrations. The analytes were stable after three freeze-thaw cycles and their dry residues were stable at -60 degrees C for 15 days. The method was successfully applied to determine concentrations of picroside-I and kutkoside post i.v. bolus administration of picroliv in rabbit. PMID- 15899376 TI - Quantification of phenolic metabolites of environmental chemicals in human urine using gas chromatography-tandem mass spectrometry and isotope dilution quantification. AB - We have developed a method to measure 12 urinary phenolic metabolites of pesticides or related chemicals. The target chemicals for our method are 2 isopropoxyphenol; 2,4-dichlorophenol; 2,5-dichlorophenol; carbofuranphenol; 2,4,5 trichlorophenol; 2,4,6-trichlorophenol; 3,5,6-trichloro-2-pyridinol; para nitrophenol, ortho-phenylphenol, pentachlorophenol, 1-naphthol and 2-naphthol. The sample preparation involves enzyme hydrolysis, isolation of the target chemicals using solid phase extraction cartridges, a phase-transfer catalyzed derivatization, cleanup using sorbent-immobilized liquid/liquid extraction cartridges, and concentration of the sample. Derivatized samples are analyzed by capillary gas chromatography-tandem mass spectroscopy using isotope dilution calibration for quantification. The limits of detection are in the mid ng/L range and the average coefficient of variation was below 15% for most of the analytes. Using our method, we measured concentrations of the target chemicals in urine samples from the general population. PMID- 15899377 TI - A highly sensitive high-performance liquid chromatography-mass spectrometry method for quantification of fludarabine triphosphate in leukemic cells. AB - A high-performance liquid chromatography (HPLC)-mass spectrometry (MS) method has been developed for the analysis of 9-beta-D-arabinofuranosyl-2-fluoroadenine 5' triphosphate (F-ara-ATP) from biological samples. Quantification is carried out by selected ion monitoring of the parent ion. Baseline separation of the monophosphate (F-ara-AMP) and diphosphate (F-ara-ADP) is achieved using the volatile ion-pairing reagent dimethylhexylamine. This method is selective and sensitive with an on-column detection limit of approximately 50 fmol. It also permits simultaneous monitoring of endogenous adenosine phosphates. The utility of the assay has been demonstrated by the analysis of F-ara-ATP in human leukemic cells after incubation with 9-beta-D-arabinosyl-2-fluoroadenine (F-ara-A) at clinically relevant concentrations. PMID- 15899378 TI - Liquid chromatography-mass spectrometry assay for quantitation of ifosfamide and its N-deschloroethylated metabolites in rat microsomal medium. AB - A specific and sensitive quantitative assay has been developed using high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC ESI-MS) for the simultaneous quantitation of the antitumor drug ifosfamide (IFM) and its two metabolites, N2-deschloroethylifosfamide (N2-DCE-IFM) and N3 deschloroethylifosfamide (N3-DCE-IFM) in microsomal medium. The analytes and the internal standard (cyclophosphamide) were isolated by ethylacetate extraction from rat liver microsomes. They were analysed on a Nucleosil C18 HD column (125 mm x 4 mm, 5 microm) using a step gradient with the mobile phase (2 mM ammonium formate and methanol). The HPLC-ESI-MS method used selected ion monitoring of ions m/z 199.1 Th and m/z 261.1 Th and was validated in the concentrations ranges of 100-5000 ng/mL for IFM and 50-2500 ng/mL for its N-deschloroethylated metabolites (DCE-IFM) with good accuracy and precision (CV less than 15%). The low limits of quantitation (LLOQ) were found at 50 ng/mL for N-deschloroethylated metabolites and at 100 ng/mL for the parent drug (IFM). The method was applied for the determination of ifosfamide and its N-deschloroethylated metabolites in rat microsomal incubations. PMID- 15899379 TI - Huperzine A--human serum albumin association: chromatographic and thermodynamic approach. AB - The synthesis of six new huperzine analogues was reported. Each product presents an amidification of the free amine on huperzine A. The synthesis strategy of these new huperzine A derivatives is based on a condensation with an acyl anhydride. The binding on HSA of two molecule series (huperzine and benzodiazepine, respectively) was investigated with high performance liquid affinity chromatography (HPLAC) using an HSA column. A thermodynamic approach showed that binding huperzine A on HSA involved hydrophobic and Van der Waals interactions. A comparative thermodynamic study with benzodiazepine molecules was carried out to determine the potential binding site of huperzine derivatives on HSA. PMID- 15899380 TI - Analysis of ameloblastomas by comparative genomic hybridization and fluorescence in situ hybridization. AB - In order to characterize the chromosomal alterations in ameloblastomas, a combination of comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) techniques was performed on 9 tumors. Chromosomal alterations including a gain at 1q and losses at 1pter, 10q, and 22q could be detected by CGH only in 1 tumor. Interphase FISH analysis, using centromeric probes for chromosomes 1, 10, and 22 as well as region-specific probes for 1p36 and 10q26, revealed the most frequent alterations to exist in the tumor with the abnormal CGH profile. These alterations included marked to slight increases of monosomic cells for chromosome 10 (91.5%), 10q26 (35.8%), 1p36 (24.4%), and chromosome 22 (18.8%), as well as significant elevations of trisomic cells for chromosome 1 (41.2%). Moreover, FISH analysis revealed a frequent loss of chromosome 22 in all tumors examined, except for one lesion, indicating that loss of the entire or a part of this chromosome is a common event in ameloblastomas, possibly being a predisposing factor to ameloblastoma tumorigenesis. PMID- 15899381 TI - Genomic deletions in cell lines derived from primitive neuroectodermal tumors of the central nervous system. AB - Extensive genomic deletions affecting a variety of chromosomes are a common finding in primitive neuroectodermal tumors of the central nervous system (CNS PNETs), implicating the loss of multiple tumor suppressor genes in the pathogenesis of these tumors. We have used representational difference analysis, microsatellite mapping, and quantitative polymerase chain reaction to identify and verify the presence of genomic deletions on a number of chromosomes in CNS PNET cell lines. This systematic approach has confirmed the importance of deletions at 10q, 16q, and 17p in PNET pathology and has revealed other regions of deletion not commonly described (e.g., Xq, 1p, 7p, and 13q). These data highlight the prevalence of hemizygous loss in CNS-PNET cells, suggesting that haploinsufficiency affecting multiple tumor suppressor genes may play a fundamental role in CNS-PNET pathogenesis. The identification of specific genes and signaling pathways that are compromised in CNS-PNET cells is crucial for development of more efficacious and less invasive treatments, as are urgently needed. PMID- 15899382 TI - High-resolution methylation analysis of the BRCA1 promoter in ovarian tumors. AB - Both hereditary and sporadic ovarian tumors frequently have decreased BRCA1 expression. One mechanism of downregulating BRCA1 expression is hypermethylation of the BRCA1 promoter. Studies have shown that the BRCA1 promoter is aberrantly hypermethylated in a subset of ovarian tumors, although the proportion varies widely between reports. High-resolution analysis of the BRCA1 promoter in ovarian cancer may provide information regarding the extent and heterogeneity of methylation and guide future studies using methylation-specific polymerase chain reaction (MS-PCR). We screened 50 primary epithelial ovarian tumors for BRCA1 promoter hypermethylation using MS-PCR. The BRCA1 promoter was hypermethylated in 16% (8 of 50) of the tumors, including two stage IA tumors. Sequence analysis of the promoter revealed that methylation of the CpG island is both extensive and mosaic in the methylated samples. Two CpG dinucleotides in the BRCA1 promoter, within and adjacent to a Myb consensus binding site, were most frequently methylated in ovarian tumors. BRCA1 expression was significantly lower in methylated than in unmethylated samples. Our analysis of the BRCA1 promoter revealed preferential methylation of specific CpG sites in ovarian tumors. This finding could be exploited in the design of highly sensitive MS-PCR assays for direct assessment of tumor DNA and potentially for early detection of ovarian cancer in body fluids. PMID- 15899383 TI - Cytogenetic aberrations in spontaneous endometrial adenocarcinomas in the BDII rat model as revealed by chromosome banding and comparative genome hybridization. AB - Female rats of the inbred strain BDII are genetically predisposed to endometrial estrogen-dependent adenocarcinomas (EAC). More than 90% of them spontaneously develop this tumor type before the age of 24 months. In order to dissect out the genetic components behind these tumors we have made crosses between BDII females and rats from 2 other strains that are nonsusceptible to EAC. It was found that EAC tumors developed in a subset of intercross and backcross animals from both interstrain crosses. The chromosomal changes in the developing tumors were studied using cytogenetic and molecular cytogenetic methods. From these studies, we conclude that certain chromosome regions were recurrently engaged in chromosomal changes such as increases in copy number (e.g., trisomy, amplification) or decreases (e.g., deletion). Based on the analysis of 56 tumors, 8 regions were found to be particularly often involved: RNO4prx, gain=34 (61%) (amplification 12 cases); RNO5mid, loss=15 (27%); RNO6prx, gain=25 (45%) (amplification 8 cases); RNO10 loss, prx-mid/gain dst=25 (45%) (amplification 1 case); RNO12q, gain=23 (41%); RNO15p loss/RNO15q gain=29 (52%) (amplification 1 case) [RNO, rat chromosome; prx, proximal; mid, middle; dst, distal; p, short arm; q, long arm]. We begun to analyze these regions in detail using various molecular methods and within them there are certain possible target genes, such as MET (RNO4q21), CDKN2A/2B (RNO5q32), MYCN (RNO6q15 approximately q16), and TP53 (RNO10q24 approximately q25), but it is clear that several other genes, still unidentified, must also be involved. PMID- 15899384 TI - Acute promyelocytic leukemia with PML-RARA fusion on i(17q) and therapy-related acute myeloid leukemia. AB - We describe a patient with acute promyelocytic leukemia (APL) and the karyotype 46,XX,i(17)(q10) with PML-RARA fusion gene detected by fluorescence in situ hybridization (FISH) and nested reverse transcriptase-polymerase chain reaction (RT-PCR). FISH using dual-color translocation probes for PML (promyelocytic leukemia) and RARA (retinoic acid receptor-alpha) showed fusion signal for PML RARA on both arms of i(17q). The patient attained complete remission (CR) with all-trans retinoic acid treatment and became PML-RARA negative. One year later, while PML-RARA negative on FISH and RT-PCR, the patient presented with thrombocytopenia. Bone marrow examination suggested an acute monoblastic leukemia (AML-M5a) including the karyotype 46,XX,t(8;16) (p11.2;p13.3),inv(11)(p15q22 approximately q23)[11]/47,idem,+i(8)(q10)[9]. She is currently in CR. The occurrence of therapy related acute leukemia after successful therapy for APL is an emerging problem. PMID- 15899385 TI - Chromosomal abnormalities in bronchial epithelium from smokers, nonsmokers, and lung cancer patients. AB - The identification of individuals who are at greatest risk of developing lung cancer would greatly improve diagnosis and possibly lead to early treatment. To study the use of karyotypes for this purpose, we used short-term human bronchial epithelial (hBE) cell cultures from nonsmokers, smokers, and lung cancer patients. Twenty-five metaphases were scored for hBE cell cultures obtained from 32 patients: 8 were nonsmokers, and 24 had a history of smoking (of whom 11 had had lung cancer surgery). The number of abnormal metaphases ranged from 0 to 4 per cell culture. No overall differences in the number of abnormal metaphases were observed between nonsmokers and smokers or between lung cancer patients and non-lung cancer patients. The most commonly observed abnormalities were structural changes in chromosome 1 (six cultures), loss of chromosome 17 (six cultures), and trisomy of chromosome 20 (three cultures). These specific alterations were found almost exclusively in patients with a history of tobacco smoking. The results did not indicate that general chromosomal abnormalities are a useful marker for tobacco smoke exposure or cancer risk. PMID- 15899386 TI - TP53 codon 72 polymorphism and risk for cervical cancer in Portugal. AB - High-risk human papillomavirus are essential for the development of cervical cancer; however, TP53 is the most frequently altered tumor suppressor gene among tumors and is described as a cofactor for cervical carcinogenesis. TP53 has two common polymorphic forms encoding either proline or arginine, at position 72, and the presence of homozygous arginine has been reported as a risk factor for cervical cancer in many populations. We evaluated the effect of this TP53 polymorphism in a northern Portuguese population. We analyzed blood samples of 385 women; 20 with low-grade squamous intraepithelial lesion (SIL), 56 with high grade SIL, 164 with invasive cervical cancer, and 145 healthy controls, using allele specific-polymerase chain reaction methodology. We observed an increased frequency of the Arg/Arg genotype in the cancer group, but no statistical significance was found between cases and controls (P>0.05). Our results indicate that there is no association between the presence of the Arg allele in codon 72 of TP53 polymorphism and risk of cervical cancer in our population. PMID- 15899387 TI - Genotoxicity and sister chromatid exchange in patients with myelodysplastic disorders. AB - In this study, we aimed to evaluate genotoxicity by sister chromatid exchange frequency in the peripheral lymphocytes of patients with myelodysplastic syndrome (MDS). The study population consisted of 16 patients (females/males:6/10; mean age: 61.68 +/- 13.08 years) diagnosed with "refractory anemia" according to FAB classification. The results were compared with an age- and sex-matched control group. The sister chromatid exchange frequency in the study group was found to be significantly higher than the control group (8.3 +/- 11 vs. 6.83 +/- 1.07, P=0.0046). We suggest that increased DNA damage, which is revealed by the increased sister chromatid exchange in the present study may play a role in the etiopathogenesis of MDS. PMID- 15899388 TI - Trisomy 8 as the sole cytogenetic abnormality in a case of extraskeletal mesenchymal chondrosarcoma. AB - Mesenchymal chondrosarcoma is a rare malignant tumor that comprises about 3-10% of all sarcomas. Reports of cytogenetic studies of mesenchymal chondrosarcoma are limited and no consistent cytogenetic abnormality has surfaced. Some mesenchymal chondrosarcomas have a t(11;22) translocation suggesting a relationship with the PNET/Ewing tumor family. We report what to our knowledge is the first case of trisomy 8 as the sole cytogenetic abnormality in a mesenchymal chondrosarcoma. PMID- 15899389 TI - Identification of a transcriptionally active hVH-5 pseudogene on 10q22.2. AB - Mitogen-activated protein kinases (MAPKs) are important regulators of a vast number of biological functions that affect life and death of eukaryotic cells and are tightly regulated by the concerted action of several phosphatases. Among these is the human homologue of vaccinia virus H1 phosphatase gene clone 5 (hVH 5) product, which dephosphorylates and thus inhibits members of the MAPK family. Here, we analyzed hVH-5 transcripts in mammary carcinoma cell lines and discovered a sequence with 88% similarity to hVH-5 transcripts. Because this variant of hVH-5 lacked intronic sequences in its genomic structure, we suggest it might be a processed pseudogene of hVH-5. psihVH-5 transcripts were detected in human peripheral tissues as well as in 11 of 14 breast cancer cell lines. In respect to the normal hVH-5 gene, the pseudogene contains several point mutations and a frame shift due to the deletion of 2 bases that would lead to the truncation of the putative psihVH-5 product. PMID- 15899390 TI - Cytogenetic characterization of Ewing tumors with high-ploidy. AB - The formation of a high-ploidy is rare in Ewing tumor (ET) and is in contrast to some other childhood tumors such as neuroblastoma. In a series of 37 Ewing tumors analyzed by conventional cytogenetics, 4 of the 34 tumors with an abnormal clone (11.8%) demonstrated the presence of a high-ploidy clone, with a chromosome number that ranged from hypotriploid to pentaploid. All 4 contained a t(11;22)(q24;q12) and the karyotypes had further aberrations of the type that would be generally expected in ET. Numerical aberrations represented the majority of the karyotypic events identified and gain of chromosome 8 and loss of chromosomes 3, 10, 16, 19, and 22 occurred in at least 3 tumors. However, no single mechanism could be implicated to explain the karyotypic picture of the 4 cases. It is proposed that high-ploidy subgroups exist in ET and it would be potentially erroneous to group these and other cases together when determining their clinical implications. PMID- 15899391 TI - BCR-ABL gene amplification and overexpression in a patient with chronic myeloid leukemia treated with imatinib. AB - Imatinib mesylate was designed as an inhibitor targeting the BCR-ABL tyrosine kinase, the molecular counterpart of the Philadelphia translocation t(9;22)(q34;q11). We report on a patient with chronic myeloid leukemia (CML) undergoing acceleration during imatinib treatment. Cytogenetic analysis revealed four different cell populations: 46,XX,t(9;22)(q34;q11),der(18)t(2;18)(p11;p11)[1]/47,idem,i(17)(q10), der(18)t(2;18),+der(22)t(9;22)[1]/46,idem, t(9;22),der(9)t(9;22),ider(22)t(9;22)[12]/ 47,idem, t(9;22),der(9)t(9;22),+22,ider(22)t(9;22)x2[1]. FISH analysis confirmed the presence of these four clones. Moreover, 49% of the interphase nuclei contained either one or two clustered fusion signals, indicating a low-level amplification of the BCR-ABL fusion gene. With quantitative real-time RT-PCR, a BCR-ABL/G6PDH ratio of 0.8 was determined, which is comparable to that measured in the K562 cell line with a known BCR-ABL amplification and which is increased by more than about 60-fold compared to a CML at diagnosis with >80% Philadelphia-positive cells. We give further evidence that the genomic BCR-ABL amplification results in an increased level of BCR-ABL transcript linking two potent mechanisms of resistance against imatinib treatment. PMID- 15899392 TI - Two cases of acute myeloid leukemia with t(11;17) associated with varying morphology and immunophenotype: rearrangement of the MLL gene and a region proximal to the RARalpha gene. AB - This report describes 2 cases of acute myeloid leukemia (AML), which based on the WHO classification would be classified as AML with an 11q23 (MLL) abnormality, but with contrasting morphologic and immunophenotypic profiles. One case had monocytic features (morphologically and immunophenotypically) with a t(11;17)(q23;q21), a previously identified variant translocation in acute promyelocytic leukemia (APL). The second case had morphologic and immunophenotypic features of APL associated with a t(11;17)(q23;q25). In both cases, fluorescence-in-situ hybridization (FISH) analysis demonstrated that the 11q23 breakpoint involved the MLL gene, but RARalpha was not involved in the 17q breakpoints. These cases illustrate the importance of FISH analysis to confirm the presence of a particular recurring rearrangement. PMID- 15899393 TI - Unique three-way translocation, t(3;14;18)(q27;q32;q21), in follicular lymphoma. AB - A 43-year-old woman was diagnosed as having stage IV follicular lymphoma. Phenotypically, the lymphoma cells were CD5(-), CD10(+), CD19(+), CD20(+), CD23( ), HLA-DR(+), and IgM-lambda(+). Conventional chromosomal analysis showed a three way t(3;14;18)(q27;q32;q21) in the lymphoma cells, which was confirmed by spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH). Immunohistochemistry revealed that both BCL2 and BCL6 proteins were expressed in the lymphoma cells, whereas only the BCL6 gene, and not the BCL2 gene, was rearranged by Southern blotting. The patient received combination chemotherapy and has been well for 3 years. This is the first reported case showing a three way translocation involving 2 major lymphoma-specific abnormalities, 3q27 and t(14;18)(q32;q21). PMID- 15899394 TI - Cytogenetic and molecular cytogenetic studies of a variant of t(21;22), ins(22;21)(q12;q21q22), with a deletion of the 3' EWSR1 gene in a patient with Ewing sarcoma. AB - Ewing sarcoma is the second most common malignant bone tumor in children and young adults. Cytogenetic analysis to identify a common t(11;22)(q23;q12) or less frequently a t(21;22)(q22;q12) or t(7;22)(p22;q12) plays an important role in the confirmation of the clinical diagnosis. We report a case of a 10-year-old female who had extraskeletal Ewing sarcoma. Conventional cytogenetic analysis revealed that 11 out of 20 cells had a derivative chromosome 22, possibly due to an insertion of the long arm of the 21q21 approximately q22. This finding was confirmed by fluorescence in situ hybridization (FISH) utilizing whole chromosome paint probes specific for chromosomes 21 and 22. Hybridization utilizing LSI EWSR1, dual-color break-apart rearrangement probe unexpectedly revealed that the 3' EWSR1 gene was lost on the derivative chromosome 22. This finding suggests that the insertion of chromosome 21 is another mechanism that could lead to EWS ERG gene fusion. To our knowledge, this is the first case report of an insertion of a segment of 21q21 approximately q22 into the long arm of 21q12 with a loss of a DNA segment around the breakpoint on the derivative chromosome 22 in Ewing sarcoma. PMID- 15899395 TI - A case of infantile acute myelogenous leukemia with MLL-MLL10 fusion caused by insertion of 11q into 10p. PMID- 15899396 TI - A t(2;3)(q11;q27) involving the BCL6 gene in follicular lymphoma with dup(12q) and t(14;18). PMID- 15899397 TI - Biphenotypic expression in a case of acute leukemia with pericentric inv(6)(p12q24). PMID- 15899398 TI - Clonal abnormalities in a recurrent laryngeal papilloma. PMID- 15899399 TI - Monosomy 22 and trisomy 14 in a granulosa tumor metastatic to the lung 20 years after the removal of the primary tumor. PMID- 15899400 TI - Fluorescence in situ hybridisation (FISH)--the next generation. AB - Fluorescence in situ hybridisation (FISH) has become one of the major techniques in environmental microbiology. The original version of this technique often suffered from limited sensitivity due to low target copy number or target inaccessibility. In recent years there have been several developments to amend this problem by increasing signal intensity. This review summarises various approaches for signal amplification, focussing especially on two widely recognised varieties, tyramide signal amplification and multiply labelled polynucleotide probes. Furthermore, new applications for FISH are discussed, which arise from the increased sensitivity of the method. PMID- 15899401 TI - Biotin biosynthesis, transport and utilization in rhizobia. AB - Biotin, a B-group vitamin, performs an essential metabolic function in all organisms. Rhizobia are alpha-proteobacteria with the remarkable ability to form a nitrogen-fixing symbiosis in combination with a compatible legume host, a process in which the importance of biotin biosynthesis and/or transport has been demonstrated for some rhizobia-legume combinations. Rhizobia have also been used to delimit the biosynthesis, metabolic effects and, more recently, transport of biotin. Molecular genetic analysis shows that an orthodox biotin biosynthesis pathway occurs in some rhizobia while others appear to synthesize the vitamin using alternative pathways. In addition to its well established function as a prosthetic group for biotin-dependent carboxylases, we are beginning to delineate a role for biotin as a metabolic regulator in rhizobia. PMID- 15899402 TI - The Pseudomonas aeruginosa pirA gene encodes a second receptor for ferrienterobactin and synthetic catecholate analogues. AB - Actively secreted iron chelating agents termed siderophores play an important role in the virulence and rhizosphere competence of fluorescent pseudomonads, including Pseudomonas aeruginosa which secretes a high affinity siderophore, pyoverdine, and the low affinity siderophore, pyochelin. Uptake of the iron siderophore complexes is an active process that requires specific outer membrane located receptors, which are dependent of the inner membrane-associated protein TonB and two other inner membrane proteins, ExbB and ExbC. P. aeruginosa is also capable of using a remarkable variety of heterologous siderophores as sources of iron, apparently by expressing their cognate receptors. Illustrative of this feature are the 32 (of which 28 putative) siderophore receptor genes observed in the P. aeruginosa PAO1 genome. However, except for a few (pyoverdine, pyochelin, enterobactin), the vast majority of P. aeruginosa siderophore receptor genes still remain to be characterized. Ten synthetic iron chelators of catecholate type stimulated growth of a pyoverdine/pyochelin deficient P. aeruginosa PAO1 mutant under condition of severe iron limitation. Null mutants of the 32 putative TonB-dependent siderophore receptor encoding genes engineered in the same genetic background were screened for obvious deficiencies in uptake of the synthetic siderophores, but none showed decreased growth stimulation in the presence of the different siderophores. However, a double knock-out mutant of ferrienterobactin receptor encoding gene pfeA (PA 2688) and pirA (PA0931) failed to be stimulated by 4 of the tested synthetic catecholate siderophores whose chemical structures resemble enterobactin. Ferric-enterobactin also failed to stimulate growth of the double pfeA-pirA mutant although, like its synthetic analogues, it stimulated growth of the corresponding single mutants. Hence, we confirmed that pirA represents a second P. aeruginosa ferric-enterobactin receptor. The example of these two enterobactin receptors probably illustrates a more general phenomenon of siderophore receptor redundancy in P. aeruginosa. PMID- 15899403 TI - Novel target genes of PsrA transcriptional regulator of Pseudomonas aeruginosa. AB - The PsrA transcriptional regulator is involved in stationary phase induced transcriptional regulation of rpoS and in negative auto-regulation in Pseudomonas aeruginosa. This study was designed to determine whether other loci were regulated by PsrA in P. aeruginosa. Computer search was performed of the PsrA binding motif (G/CAAAC N(2-4) GTTTG/C) against the P. aeruginosa genome sequence. Four of 14 analysed promoters responded to and bound PsrA; (i) divergent promoters controlling PA2952/PA2951 and PA2953, (ii) promoter of PA0506 and (iii) upstream region of PA3571. Promoters PA0506 and PA2952-PA2951 were regulated negatively whereas promoters of PA2953 and PA3571 were regulated positively by PsrA. Two dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (2D SDS-PAGE) analysis on total proteins from P. aeruginosa PAO1 and psrA knock out derivative was also performed resulting in the identification of 11 protein spots which were differentially regulated. These studies have indicated PsrA as a global regulator. PMID- 15899404 TI - Isolation and characterisation of the lipopolysaccharide from Acidiphilium strain GS18h/ATCC55963, a soil isolate of Indian copper mine. AB - The lipopolysaccharide (LPS) of the Gram-negative Acidiphilium strain GS18h/ATCC55963, a new soil isolate, exhibited very low endotoxic activity as determined by Limulus gelation activity, lethal toxicity in galactosamine (GalN) sensitised mice, and level of tumor necrosis factor alpha (TNFalpha) in the blood serum of BALB/c mice. Analysis of the LPS, specially of lipid A which usually accounts for the toxicity, revealed the latter to contain glucosamine and phosphate besides fatty acids, of which 14:0(3-OH), 18:0(3-OH), 18:1 and 19:0(cyclo) are the major components, while 12:0, 16:0, 19:1, 20:0(3-OH) and 20:1(3-OH) are present in small amounts. The 14:0(3-OH) and 18:0(3-OH) fatty acids are amide-linked, whereas the rest are ester bound. Glucose, galactose, mannose, rhamnose, heptose, galacturonic acid and 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) were present in the polysaccharide part of this LPS. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the LPS showed a macromolecular heterogeneity distinctly different from those of Escherichia coli or Salmonella. The toxicity of this LPS being extremely low attributed to fatty acid composition of its lipid A, promises potential therapeutic application. PMID- 15899405 TI - Comprehensive analysis of classical and newly described staphylococcal superantigenic toxin genes in Staphylococcus aureus isolates. AB - We describe a comprehensive detection system for 18 kinds of classical and newly described staphylococcal superantigenic toxin genes using four sets of multiplex PCR. Superantigenic toxin genotyping of Staphylococcus aureus for 69 food poisoning isolates and 97 healthy human nasal swab isolates revealed 32 superantigenic toxin genotypes and showed that many S. aureus isolates harbored multiple toxin genes. Analysis of the relationship between toxin genotypes and toxin genes encoding profiles of mobile genetic elements suggests its possible role in determining superantigenic toxin genotypes in S. aureus as combinations of toxin gene-encoding mobile genetic elements. PMID- 15899406 TI - Passive immunisation of hamsters against Clostridium difficile infection using antibodies to surface layer proteins. AB - Clostridium difficile is a major cause of antibiotic-associated diarrhoea and the primary cause of pseudomembraneous colitis in hospitalised patients. We assessed the protective effect of anti-surface layer protein (SLP) antibodies on C. difficile infection in a lethal hamster challenge model. Post-challenge survival was significantly prolonged in the anti-SLP treated group compared with control groups (P=0.0281 and P=0.0283). The potential mechanism of action of the antiserum was shown to be through enhancement of C. difficile phagocytosis. This report indicates that anti-SLP antibodies can modulate the course of C. difficile infection and may therefore merit closer investigation for use as constituents of multi-component vaccines against C. difficile associated diarrhoea. PMID- 15899407 TI - Morphological and molecular taxonomy of Pythium longisporangium sp. nov. isolated from the Burgundian region of France. AB - During the course of an investigation on the Pythiaceous oomycetes occurring in the Burgundian vineyards, some species of Pythium possessing mainly hypogynous antheridia were found. These had been classified as oomycetes belonging to the "Pythium rostratum" group for a long time. Three of these isolates, having similar structures and growth, are very closely related to a recently described species, Pythium bifurcatum Paul. A close look at these, however, underlines some fundamental differences with the latter. Not all of them produce zoospores but have very large sporangia. The type specimen is F-1200 (B 76a) which is a medium slow growing saprophyte. The sequence of the ITS region of the rDNA also shows a very close relationship with P. bifurcatum. On the basis of morphological and molecular analysis, we now describe this species as Pythium longisporangium sp. nov. Morphological features of this new species, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed. PMID- 15899408 TI - Polymorphism and gene conversion of the 16S rRNA genes in the multiple rRNA operons of Vibrio parahaemolyticus. AB - The genome sequence of a strain of Vibrio parahaemolyticus holds 11 copies of rRNA operons (rrn) with identical 16S rRNA genes (rrs). Conversely, the species type strain contains two rrs classes differing in 10 nucleotide sites within a short segment of 25 bp. Furthermore, we show here that the sequence of this particular segment largely differs between some strains of this species. We also show that of the eleven rrn operons in the species type strain, seven contain one rrs class and four the other, indicating gene conversion. Our results support the hypothesis that the rrs differences observed between strains of this species were caused by lateral transfer of an rrs segment and subsequent conversion. PMID- 15899409 TI - Induction of murine macrophage TNF-alpha synthesis by Mycobacterium avium is modulated through complement-dependent interaction via complement receptors 3 and 4 in relation to M. avium glycopeptidolipid. AB - We studied whether complement receptor (CR) mediated Mycobacterium avium interaction modulated macrophage TNF-alpha expression. Compared to control conditions, infections performed with C3-depletion yielded significantly higher TNF-alpha levels. Blockage of the CR4 iC3b site yielded increases in TNF-alpha for all morphotypic variants of a virulent serovar-8 strain (smooth transparent (SmT), smooth opaque (SmO), serovar-specific glycopeptidolipid (ssGPL) deficient knockout mutant) whereas CR3 blockage increased TNF-alpha only for SmT and ssGPL deficient strains. Thus, complement-mediated binding of M. avium to CR3 and CR4 was shown to modulate TNF-alpha expression. The differential activation of morphotypic and isogenic variants of a single strain provides an excellent model system to delineate signaling pathways. PMID- 15899410 TI - Overexpression of a hydrogenase gene in Clostridium paraputrificum to enhance hydrogen gas production. AB - A [Fe]-hydrogenase gene (hydA) was cloned from Clostridium paraputrificum M-21 in Escherichia coli using a conserved DNA sequence of clostridial hydrogenase genes amplified by PCR as the probe. The hydA gene consisted of an open reading frame of 1749 bp encoding 582 amino acids with an estimated molecular mass of 64,560 Da. It was ligated into a shuttle vector, pJIR751, originally constructed for Clostridium perfringens and E. coli, and expressed in C. paraputrificum. Hydrogen gas productivity of the recombinant increased up to 1.7-fold compared with the wild-type. In the recombinant, overexpression of hydA abolished lactic acid production and increased acetic acid production by over-oxidation of NADH, which is required for reduction of pyruvic acid to lactic acid in the wild-type. PMID- 15899411 TI - RirA is the iron response regulator of the rhizobactin 1021 biosynthesis and transport genes in Sinorhizobium meliloti 2011. AB - The genes encoding the biosynthesis and transport of rhizobactin 1021, a siderophore produced by Sinorhizobium meliloti, are negatively regulated by iron. Mutagenesis of rirA, the rhizobial iron regulator, resulted in abolition of the iron responsive regulation of the biosynthesis and transport genes. Bioassay analysis revealed that the siderophore is produced in the presence of iron in a rirA mutant. RNA analysis and GFP fusions supported the conclusion that RirA is the mediator of iron-responsive transcriptional repression of the two transcripts encoding the biosynthesis and transport genes. RirA in S. meliloti appears to fulfil the role often observed for Fur in other bacterial species. The regulator was found to mediate the iron-responsive expression of two additional genes, smc02726 and dppA1, repressing the former while activating the latter. The rirA mutant nodulated the host plant Medicago sativa (alfalfa) and fixed nitrogen as effectively as the wild type. PMID- 15899412 TI - Polymerase chain reaction for identification of aldoxime dehydratase in aldoxime- or nitrile-degrading microorganisms. AB - We developed a molecular screening procedure using Southern hybridization and polymerase chain reaction (PCR) to identify aldoxime dehydratase (Oxd) encoding genes (oxds) among 14 aldoxime- or nitrile-degrading microorganisms. When an oxd gene of Rhodococcus erythropolis N-771 was used as a probe, positive hybridization signals were seen with the chromosomal DNA of eight strains, suggesting that these strains have similar oxd genes to R. erythoropolis N-771. By analyzing the PCR-amplified fragments with degenerate consensus primers, the occurrence of homologous Oxd coexisting with Fe-containing NHase in the active eight strains was demonstrated coinciding with the results of Southern hybridization. Whole length of oxd gene was cloned as an example from one of the positive strains, Pseudomonas sp. K-9, sequenced, and expressed in E. coli. Analysis of the primary structure of the protein (OxdK) encoded by the oxd gene of Pseudomonas sp. K-9 led to identify an Oxd having a new primary structure. Thus, the PCR-based analysis of oxd gene is a useful tool to detect and analyze the "aldoxime-nitrile pathway" in nature, since Oxd is the key enzyme for the pathway. PMID- 15899413 TI - The gene encoding xylulose-5-phosphate/fructose-6-phosphate phosphoketolase (xfp) is conserved among Bifidobacterium species within a more variable region of the genome and both are useful for strain identification. AB - The nucleotide sequence of the xfp-gene region in six known and two unknown species of Bifidobacterium was determined and compared with the published sequences of B. animalis subsp. lactis DSM10140 and B. longum biovar longum NCC2705. The xfp coding sequences were 73% identical and coded for 825 amino acids in all 10 sequences. Partial sequences of an adjacent gene, guaA, were 61% identical in six sequences for which data were available. The region between xfp and guaA was variable in both length and sequence. Oligonucleotide sequences from the conserved and variable xfp regions were used as PCR primers, in combinations of appropriate specificity, for the detection and identification of Bifidobacterium isolates. PMID- 15899414 TI - The stabilization of housekeeping transcripts in Trypanosoma cruzi epimastigotes evidences a global regulation of RNA decay during stationary phase. AB - The relative steady state concentration of mRNAs of four housekeeping single-copy type Trypanosoma cruzi genes (actin, triosephosphate isomerase, trypanothion reductase and the ribosomal protein S4) was analyzed throughout the growth curve. A distinguishable pattern was observed with maximal levels occurring at the logarithmic phase of growth and minimum levels occurring at the stationary phase. The half-lives of all analyzed messenger RNAs, and also of three molecular species of immature ribosomal RNAs were increased in cells isolated from stationary phase. These results suggest the occurrence of a novel global regulation mechanism that might protect transcripts from degradation in stationary epimastigotes, probably as a strategy to perpetuate through this quiescent stage. PMID- 15899415 TI - Genotypic and phenotypic characterization of a biofilm-forming Serratia plymuthica isolate from a raw vegetable processing line. AB - Recently, we isolated from a raw vegetable processing line a Serratia strain with strong biofilm-forming capacity and which produced N-acyl-L-homoserine lactones (AHLs). Within the Enterobacteriaceae, strains of the genus Serratia are a frequent cause of human nosocomial infections; in addition, biofilm formation is often associated with persistent infections. In the current report, we describe the detailed characterization of the isolate using a variety of genotypic and phenotypic criteria. Although the strain was identified as Serratia plymuthica on the basis of its small subunit ribosomal RNA (16S rRNA) gene sequence, it differed from the S. plymuthica type strain in production of pigment and antibacterial compounds, and in AHL production profile. Nevertheless, the identification as S. plymuthica could be confirmed by gyrB phylogeny and DNA:DNA hybridization. PMID- 15899416 TI - Chemotypes significance of lichenized fungi by structural characterization of heteropolysaccharides from the genera Parmotrema and Rimelia. AB - Galactoglucomannans were isolated from the lichenized fungi of the genus Parmotrema (Parmotrema austrosinense, Parmotrema delicatulum, Parmotrema mantiqueirense, Parmotrema schindlerii, and Parmotrema tinctorum and that of Rimelia (Rimelia cetrata and Rimelia reticulata) via successive hot alkaline extraction and precipitation with Fehling solution. The structure of each polysaccharide was investigated using 13C NMR and HSQC-DEPT spectroscopy, methylation analysis, and HPSEC-MALLS. The galactoglucomannans had a (1-->6) linked main chain of alpha-Manp units, substituted preferentially at O-2 and O-4 by alpha-Galp and beta-Galp nonreducing end-units, respectively. The C-1 region of the 13C NMR spectra of these heteropolysaccharides is typical of the lichen species, and is an additional tool in lichenized fungi classification. PMID- 15899417 TI - Isolation of genes differentially expressed during the fruit body development of Pleurotus ostreatus by differential display of RAPD. AB - To analyze genes involved in fruit body development of Pleurotus ostreatus, mRNAs from three different developmental stages: i.e., vegetative mycelium, primordium, and mature fruit body, were isolated and reverse-transcribed to cDNAs. One hundred and twenty random PCR amplifications were performed with the cDNAs, which generated 382, 394, 393 cDNA fragments from each developmental stage. From these fragments, four cDNA clones specifically expressed in primordium or mature fruit body were detected. Sequence analysis and database searches revealed significant similarity with triacylglycerol lipase, cytochrome P450 sterol 14 alpha demethylase and developmentally regulated genes of other fungi. Northern blot analyses confirmed that all of the four cDNAs were unexpressed in mycelium, thus stage-specific genes for fruit body formation of P. ostreatus were successfully isolated. PMID- 15899418 TI - Is there a relationship between clear corneal cataract incisions and endophthalmitis? PMID- 15899420 TI - Biaxial phacoemulsification. PMID- 15899419 TI - Combining wavefront and topography data. PMID- 15899421 TI - Subepithelial scarring after laser-assisted subepithelial keratectomy. PMID- 15899423 TI - Pupil size and the ablation zone in laser refractive surgery. PMID- 15899425 TI - Effect of prophylactic antibiotics on antimicrobial resistance of viridans streptococci in the normal flora of cataract surgery patients. PMID- 15899426 TI - Inadvertent vitreous staining. PMID- 15899427 TI - Consultation section. Cataract surgical problem. PMID- 15899439 TI - Slitlamp biomicroscopy of the tear film of patients using topical Restasis and Refresh Endura. AB - We describe a technique in which slitlamp biomicroscopy is performed to evaluate eyes in patients using a topical dry-eye agent; ie, topical cyclosporine A ophthalmic emulsion 0.05% (Restasis) or glycerin 1%-polysorbate 80 1% (Refresh Endura). Both are delivered in an oil-emulsion formulation. We also describe a previously unreported physical finding in the tear film of these patients. The examination technique can help confirm patient compliance and the tear-film stability of the 2 agents. PMID- 15899440 TI - Intraoperative floppy iris syndrome associated with tamsulosin. AB - PURPOSE: To assess the incidence and possible causative factors of a newly recognized syndrome, the intraoperative floppy iris (IFIS). SETTING: Clinical practices in Los Altos and San Rafael, California, USA. METHODS: A retrospective chart review of consecutive cataract surgeries performed in a 2-surgeon practice over a 12-month period (706 eyes; 511 patients) was used to assess the percentage of cataract patients on systemic sympathetic alpha-1 antagonist medications as well as the percentage of patients who manifested the IFIS. A separate prospective study of 900 consecutive cases (741 patients) performed by another surgeon was used to determine the incidence of IFIS and the percentage of these patients who were taking alpha-1 antagonist medications. RESULTS: Three percent (16/511) of the patients in the retrospective study, representing 3.0% (25/706) of the total eyes, were taking tamsulosin (Flomax) for benign prostatic hypertrophy. The overall prevalence of IFIS was 2.0% (10/511 patients). The syndrome was noted intraoperatively in 63.0% (10/16) of the tamsulosin patients but in none of the 11 patients on other systemic alpha-1 blockers. In the prospective study of 900 consecutive cataract surgeries, the prevalence of IFIS was 2.2% (16/741 patients). Ninety-four percent (15/16) of the IFIS patients were taking or had taken systemic tamsulosin. Twenty-six patients (36 eyes) in the 2 studies had IFIS associated with systemic tamsulosin. Sphincterotomies and mechanical pupil stretching were ineffective in maintaining adequate pupil dilation in this surgical population. CONCLUSION: Intraoperative floppy iris syndrome occurred in approximately 2% of a cataract surgery population and appeared to be caused by tamsulosin, a systemic sympathetic alpha-1A antagonist medication that is the most frequently prescribed medication for benign prostatic hypertrophy. PMID- 15899441 TI - Pigment dispersion and Artisan phakic intraocular lenses: crystalline lens rise as a safety criterion. AB - PURPOSE: To validate the theory that crystalline lens rise can be used as a safety criterion to prevent pigment dispersion in eyes with an Artisan phakic intraocular lens (IOL) (Ophtec BV). SETTING: Monticelli Clinic, Marseilles, France. METHOD: A comparative analysis of crystalline lens rise in 9 eyes with pigment dispersion and 78 eyes without dispersion was performed. All eyes had previous implantation of an Artisan IOL. Anterior segment imaging was done using an anterior chamber optical coherence tomography (AC OCT) prototype. Crystalline lens rise was defined by the distance between the anterior pole of the crystalline lens and the horizontal plane joining the opposite iridocorneal recesses. RESULTS: The study confirmed that crystalline lens rise can be considered a safety criterion for implantation of Artisan-type phakic IOLs. The higher the crystalline lens rise, the greater the risk for developing pigment dispersion in the area of the pupil. This complication occurred more frequently in hyperopic eyes than in myopic eyes. Results indicate there is little or no risk for pigment dispersion if the rise is less than 600 microm; 67% of eyes with a rise of 600 microm or more developed pupillary pigment dispersion. In some cases in which the IOL was loosely fixated, there was no traction on the iris root and dispersion was prevented or delayed. CONCLUSIONS: Crystalline lens rise should be considered a new safety criterion for Artisan phakic IOL implantation and should also be applied to other types of phakic IOLs. The distance remaining between the crystalline lens rise and a 600 microm theoretical safety level allows one to calculate how long the IOL can safely remain in the eye. PMID- 15899442 TI - Masked comparison of silicone hydrogel lotrafilcon A and etafilcon A extended wear bandage contact lenses after photorefractive keratectomy. AB - PURPOSE: To compare the efficacy of 2 types of bandage contact lenses (BCLs) after photorefractive keratectomy (PRK). SETTING: Navy Refractive Surgery Center, Naval Medical Center San Diego, San Diego, California, USA. METHODS: In this prospective study, 100 patients received PRK in both eyes. Each patient received a BCL composed of etafilcon A (Acuvue [ACV], Vistakon Inc.) (14.0 diameter, 8.4/8.8 base curve) in 1 eye and lotrafilcon A (Focus Night & Day [N&D], Ciba Vision) (13.8 diameter, 8.4/8.6 base curve) in the fellow eye. The patient was masked to the lens type in each eye. The postoperative medication regimen was the same with both lenses. The epithelial defect size and subjective level of discomfort were measured at surgery and daily after surgery until both eyes had reepithelialized and the lenses were removed. RESULTS: The mean epithelial defect size at surgery was similar with both BCLs (ACV 57.07 mm(2) and N&D 57.53 mm(2); P=.422). On postoperative days 1 and 2, the mean defect size was significantly smaller in eyes with the N&D lens (day 1, ACV 21.53 and N&D 18.74; day 2, ACV 3.62 and N&D 2.12) (paired t test, P<.001). On day 1, no eye had completely reepithelialized; on day 2, 20 eyes with N&D lenses and 13 eyes with ACV lenses had. On day 3, 70 eyes with N&D lenses and 66 eyes with ACV lenses had reepithelialized. The mean discomfort index was significantly higher in the eyes with ACV lenses on days 1 and 2 (paired t test, P<.001 and P<.002, respectively). Topical nonpreserved tetracaine was used significantly more often in eyes with ACV lenses on days 1, 2, and 3 (paired t test, P<.001, P<.008, P<.003, respectively). No correlation between the use of tetracaine in the first 24 hours and the rate of reepithelialization was noted (R(2)=0.0025 for ACV and R(2)=0.0003 for N&D). CONCLUSION: The lotrafilcon A lens resulted in significantly faster corneal reepithelialization and reduced patient discomfort in most patients during the first 48 hours after PRK. PMID- 15899443 TI - Visual field changes after laser in situ keratomileusis. AB - PURPOSE: To determine whether laser in situ keratomileusis (LASIK) affects the central 30-degree visual field. SETTING: University-based ophthalmology practice. METHODS: This nonrandomized clinical trial comprised 14 normal patients (27 eyes) scheduled to have LASIK for myopia or myopic astigmatism. Automated static perimetry was performed before and 6 months after surgery using the Octopus 1-2-3 perimeter and the Dynamic-32 test strategy. Patient data included sex, age, preoperative and postoperative refractive errors, preoperative and postoperative best corrected visual acuity, preoperative corneal thickness, programmed optical zone, programmed total ablation diameter, and duration of microkeratome suction. All surgery was performed using the same Alcon LADARVision 4000 excimer laser. The main outcome measures were the mean sensitivity (MS) change in the central 15 degree visual field and the MS change in the 15- to 30-degree visual field. A multivariate analysis of the MS change as a function of preoperative clinical parameters was performed. RESULTS: There was no significant change in the MS in the central 15-degree visual field; between 15 and 30 degrees, there was a statistically significant decrease of -0.82 dB +/- 1.40 (SD) (P=.01, 2-tailed t test). The decline in MS was positively correlated with refractive error and corneal thickness; it was negatively correlated with the programmed optical zone diameter. CONCLUSIONS: Automatic static perimetry can detect decreased sensitivity in the midperipheral visual field after myopic LASIK. It may be a useful quantitative subjective test for measuring the effects of future improvements in surgical technique on vision quality. PMID- 15899444 TI - Comparison of tropicamide and cyclopentolate for cycloplegic refractions in myopic adult refractive surgery patients. AB - PURPOSE: To compare tropicamide 1%, a shorter-acting cycloplegic agent, with cyclopentolate 1% for cycloplegic refractions in adult refractive surgery patients. SETTING: Navy Refractive Surgery Center, Ophthalmology, Naval Medical Center, San Diego, California. METHODS: The study was prospective, single center, with randomized sequencing of cycloplegic agent; each patient received both agents. Thirty consecutive myopic adult refractive surgery patients (mean age 35.4 years) participated. A complete preoperative examination, including cycloplegic refraction, was obtained twice, 1 week apart. The patient and the examiner were masked to the medication. Main outcome measures included cycloplegic and manifest refractions, best corrected distance acuity, near-point accommodation, pupil diameters, and subjective appraisal of experience with cycloplegic agents. RESULTS: Twenty-eight of 30 patients completed both examinations. Both eyes were measured, but comparisons were limited to right and left eyes, independently. No statistically significant difference was found between the tropicamide and cyclopentolate cycloplegic refractions (mean difference in MSE +/- SD, OD=0.054 +/- 0.214 diopters (D), t=1.33, P=.10; OS=0.054 +/- 0.253 D, t=1.12, P=.14). Five eyes of 3 patients had a difference of 0.50 D or greater between the 2 agents; less myopia with cyclopentolate. Near point testing revealed less residual accommodation with cyclopentolate (difference in MSE, OD=-0.27 +/- 0.51 D, t=2.68, P=.006; OS=-0.32 +/- 0.49 D, t=3.46, P=.001). Subjectively, 24 of 28 (86%) patients preferred tropicamide, 1 (4%) preferred cyclopentolate, and 3 (10%) had no preference. CONCLUSIONS: There was no statistically significant difference in mean cycloplegic refractions. Cyclopentolate was more effective than tropicamide in reducing accommodative amplitude in adult myopes (near-point testing). Patients strongly preferred tropicamide. PMID- 15899445 TI - Pachymetric evaluation prior to laser in situ keratomileusis. AB - PURPOSE: To determine whether deviations in the localization of the cornea's thinnest point or the magnitude and localization of posterior corneal ectasia is associated with deviations in the spherical equivalent, the astigmatism, or the magnitude of an anterior corneal ectasia and whether corneas at risk for iatrogenic keratectasia can be identified without a pachymetry map of the cornea. SETTING: University hospital eye clinic. METHODS: Three hundred eight eyes of 156 healthy volunteers with various refractive errors were examined with Orbscan II and autorefractometer-keratometer. The corneal thickness was registered at the fixation point, at the geometrical center, and at the thinnest point of the cornea. Keratometry and refraction were determined for all subjects. RESULTS: The thinnest point of the cornea was predominantly located in the inferotemporal quadrant, and was significantly thinner than the fixation point (539.6 +/- 35.8 microm and 548.0 +/- 35.4 microm, respectively, P<.001). Interestingly, the larger this difference was, the longer the distance between these points. No relationship was found between the refractive or external surface measurements and the internal surface measurements. CONCLUSIONS: The absence of a clear relationship between the shape of the anterior corneal surface or the refractive error, and the shape of the posterior corneal surface, necessitates a thorough pachymetric evaluation of the cornea before a laser in situ keratomileusis procedure, with special attention to the inferotemporal area. PMID- 15899446 TI - Assessment of central corneal thickness using optical coherence tomography. AB - PURPOSE: To demonstrate the capability of model OCT3 optical coherence tomographer to evaluate central corneal thickness (CCT) in normal human corneas in vivo and compare the results with the those of standard ultrasound (US) pachymetry and Orbscan. SETTING: New York Eye & Ear Infirmary Resident Clinic, New York, New York, USA. METHODS: The CCT in 22 eyes of 11 subjects was determined with the OCT3 (Carl Zeiss Meditec), Orbscan (Bausch & Lomb, Inc.), and US pachymetry (DGH Technology, Inc.). Three central corneal scans of each eye were obtained using the OCT3. First, OCT3 data were processed using the standard OCT software program (OCT3(std)). Second, OCT3 raw data were exported and measurements were repeated using Scion Image for Windows program (OCT3(sci)). The OCT3 and Orbscan results were compared with the mean of 5 US pachymetry measurements in each eye. RESULTS: The OCT3(std), OCT3(sci), and Orbscan CCT measurements showed high correlations with US pachymetry (r=0.981, r=0.984, and r=0.942, respectively; P<.0001). Bland-Altman analysis showed a high level of agreement between US pachymetry and OCT3 techniques but not Orbscan. High repeatability for OCT3(std) (r(2)=0.05) and OCT3(sci) (r(2)=0.01) was also seen. CONCLUSION: Results show the OCT3 is an accurate, noninvasive, and reproducible technique for evaluation of CCT. PMID- 15899447 TI - Visual acuity and contrast sensitivity comparison between Tecnis and AcrySof SA60AT intraocular lenses: A multicenter randomized study. AB - PURPOSE: To evaluate best corrected visual acuity (BCVA) and photopic and mesopic contrast sensitivity in pseudophakic patients implanted either with the aspheric intraocular lens (IOL) designed to correct for corneal spherical aberration or with a conventional IOL. SETTING: Three surgical centers participated this prospective randomized masked comparative study. METHODS: Thirty eyes of 30 patients after aspheric lens implantation (Pfizer/Pharmacia Tecnis Z9000) were compared with 30 eyes of 30 age-matched patients after conventional lens implantation (Alcon AcrySof SA60AT). Two to three months after surgery, best spectacle corrected distance visual acuity was measured using the Early Treatment Diabetic Retinopathy Study chart. Contrast sensitivity was measured by sinusoidal grating charts for distance at photopic (85 cd/m(2)) and mesopic (6 cd/m(2)) luminance level with optical correction in place. Tested spatial frequencies were 1.5, 3, 6, 12, and 18 cycles per degree (cpd). RESULTS: The mean BCVA was -0.053 +/- 0.044 logMAR in eyes with the aspheric Tecnis IOL and 0.006 +/- 0.059 logMAR in eyes with the conventional AcrySof IOL (P=.0001). Eyes with the aspheric IOL showed better contrast sensitivity at spatial frequencies of 3 cpd (P<.05) and 6, 12, and 18 cpd (P<.01) in photopic and in mesopic conditions. Peak improvement occurred at 18 cpd and was 36% (0.29 log units) in photopic and 54% (0.27 log units) in mesopic conditions. CONCLUSIONS: The aspheric Tecnis IOL yielded better BCVA and better distance contrast sensitivity than the conventional IOL. The differences were clinically significant for higher spatial frequencies. The results give some suggestions for further studies. PMID- 15899448 TI - Comparison of posterior capsule opacification rates between hydrophilic and hydrophobic single-piece acrylic intraocular lenses. AB - PURPOSE: To determine the effect of intraocular lens (IOL) material on the development of posterior capsule opacification (PCO) at 1 year. SETTING: Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. METHODS: One hundred six eyes of 53 patients with bilateral cataract and no other ocular comorbidity were prospectively randomized to receive a hydrophobic acrylic or hydrophilic acrylic single-piece IOL in the first eye to have surgery. The alternate IOL was implanted in the fellow eye 4 to 6 weeks later. All surgery was performed by a single surgeon. Postoperative follow-up was 1 day, 1 and 6 months, and 1 year. At each visit, the best corrected high- and low-contrast visual acuities were assessed and a high-intensity digital retroillumination photograph was taken. Posterior capsule opacification was assessed from the digital images by a single operator using a dedicated software program and calculated as the percentage area of opacified capsule. RESULTS: One year postoperatively, the median percentage area of PCO was 50.3% in the hydrophilic IOL group and 4.9% in the hydrophobic IOL group (P<.001). The difference in PCO was not accounted for by loss of contact between the capsulorhexis and IOL surface. Further analysis showed that lens epithelial cells tended to invade the posterior capsule at the haptic-optic junction. This was more marked in the hydrophilic IOL group. CONCLUSIONS: The rate of PCO was significantly higher with the hydrophilic IOL. However, the results cannot be attributed to the IOL material alone as they show the importance of both IOL material and design. PMID- 15899449 TI - Effect of cataract surgery incision location and intraocular lens type on ocular aberrations. AB - PURPOSE: To determine whether Hartmann-Shack wavefront sensing detects differences in optical performance in vivo between poly(methyl methacrylate) (PMMA) and foldable acrylic intraocular lenses (IOLs) and between clear corneal and scleral tunnel incisions and whether optical differences are manifested as differences in visual performance. SETTING: Department of Optometry, University of Bradford, West Yorkshire, United Kingdom. METHODS: This study comprised 74 subjects; 17 were phakic with no ocular pathology, 20 had implantation of a Pharmacia 722C PMMA IOL through a scleral tunnel, 21 had implantation of an Alcon AcrySof IOL through a scleral tunnel, and 16 had implantation of an AcrySof IOL through a corneal incision. Visual acuity and contrast sensitivity testing, ocular optical quality measurement using Hartmann-Shack wavefront sensing, and corneal surface measurement with a videokeratoscope were performed in all cases. RESULTS: There were significant differences between groups in the total root-mean square (RMS) wavefront aberration over a 6.0 mm pupil (F=3.91; degrees of freedom=3,70; P<.05) mediated at the 4th-order RMS, specifically spherical and tetrafoil aberrations. The PMMA-scleral group had the least aberrations and the AcrySof-corneal group the most. For a 3.5 mm diameter pupil, the total higher order RMS wavefront aberration was not significantly different between the groups (P>.05). There were no differences between groups in corneal shape, visual acuity, or contrast sensitivity. CONCLUSIONS: Implantation of the spherical PMMA IOL led to a slight reduction in total wavefront aberration compared to phakic eyes. AcrySof IOLs induced more aberrations, especially spherical aberration. Corneal-based incisions for IOL implantation compounded this increase. Studies of the optical performance of IOLs in vivo should use wavefront sensing as the main outcome measure rather than visual measures, which are readily confounded by multiple factors. PMID- 15899450 TI - Cohort study of 27 cases of endophthalmitis at a single institution. AB - PURPOSE: To identify potential risk factors associated with post-cataract surgery bacterial endophthalmitis. SETTING: The John A. Moran Eye Center, Salt Lake City, Utah, USA. METHODS: This retrospective cohort study consisted of patients who had surgery for cataract(s) at this eye hospital. A 10% sampling of all patients operated on for cataract surgery from January 1, 1996, to December 31, 2002, were compared with all cases of postcataract surgery bacterial endophthalmitis during this same time period at this institution. The main outcome measure(s) included surgical complication, first postoperative day wound leak, incision placement and location, intraocular lens material, whether a suture was placed, antibiotic used, collagen shield use, and whether the eye was patched. RESULTS: A total of 1525 patients were in the control cohort, and there were 27 cases of endophthalmitis. In a multivariate regression analysis, the factors found to be statistically associated with endophthalmitis were (1) wound leak on the first postoperative day (odds ratio [OR] 44 +/- 42; confidence interval [CI] 6.85 to 287; P<.001); (2) capsular or zonular surgical complication (OR 17.2 +/- 14.2; CI 3.44 to 86.4; P=.001); (3) topical antibiotic started the day after surgery rather than the day of surgery (OR 13.7 +/- 12.9; CI 2.17 to 90.9; P=.005); (4) use of ciprofloxacin rather than ofloxacin topically after surgery (OR 5.3 +/- 3.6; CI 1.41 to 20.0; P=.014); (5) not patching after surgery (OR 7.1 +/- 5.6; CI 1.47 to 36.4; P=.015); and (6) not placing a collagen shield soaked in antibiotic (OR 2.7 +/- 1.3; CI 1.06 to 7.14 P=.037). CONCLUSION: In sutureless cataract surgery, surgical complications and wound leak on the first postoperative day were most strongly associated with endophthalmitis. PMID- 15899451 TI - Corneal endothelial cell change after phacoemulsification relative to the severity of diabetic retinopathy. AB - PURPOSE: To investigate the differences in corneal endothelial cell morphology between diabetic patients who were divided by the degree of the severity of diabetic retinopathy and normal patients after phacoemulsification with intraocular lens (IOL) implantation. SETTING: Department of Ophthalmology, College of Medicine, Pusan National University, Pusan, Korea. METHODS: Before and 6 months after phacoemulsification, specular microscopy was used to evaluate the number and morphology of endothelial cells in patients with diabetic retinopathy. The patients were divided into 3 groups: group I (n=30 eyes), without diabetes; group II (n=30 eyes), diabetes with severe nonproliferative diabetic retinopathy; group III (n=30 eyes), diabetes with high-risk proliferative diabetic retinopathy (PDR). RESULTS: Postoperative corneal endothelial cell density and hexagonality were significantly decreased and the coefficient of variation in cell size increased in all groups, in contrast to the preoperative state (P<.05). During the postoperative 6 months, the percentage of hexagonal cells showed a tendency to decrease progressively, and this was a significant difference between the 3 groups (P<.05). However, the patterns of change in endothelial cell density and the coefficient of variation in cell size were not different between the groups (P>.05). At the postoperative 6 months, the corneal endothelial cell density and the coefficient of variation in cell size of high-risk PDR patients were statistically changed compared with normal persons (P<.05). However, the percentage of hexagonal cell was not significantly difference between diabetic retinopathy patients and normal patients (P>.05). CONCLUSIONS: Corneal endothelial cell density significantly decreased and the coefficient of variation in cell size significantly increased for high-risk PDR patients undergoing phacoemulsification, in contrast to normal persons at the postoperative 6 months. PMID- 15899452 TI - Congenital cataract extraction with primary aphakia and secondary intraocular lens implantation in the ciliary sulcus. AB - PURPOSE: To evaluate the results of cataract extraction without primary intraocular lens (IOL) implantation in children. SETTING: Ophthalmology Unit, University Hospital, Strasbourg, France. METHODS: This retrospective study comprised 157 congenital cataract cases (55 bilateral and 47 unilateral) treated between 1985 and 2001. Evaluated were the functional results (visual acuity, binocular vision) and factors influencing the prognosis (age at surgery, stage of cataract development, associated pathology, postoperative complications). In all patients, cataract extraction was via the pars plana and anterior vitrectomy was performed, leaving a peripheral capsular collarette in place. Postoperatively, all the patients were fitted with glasses or contact lenses, after which they had secondary implantation of an IOL in the ciliary sulcus. RESULTS: The functional results were similar to those in the literature for eyes with or without an IOL. Nevertheless, the literature reports a 25% risk for reoperation in the first 2 years after implantation and secondary vitrectomy for reopacification of the visual axis in 20% of bilateral cases and 38% of unilateral cases. CONCLUSIONS: Our functional results indicate that in cases of bilateral congenital cataracts, initial rehabilitation with aphakic correction and secondary IOL implantation leads to a predictable postoperative refraction and fewer complications. Visual rehabilitation in unilateral aphakia was more difficult because of poor compliance with contact lenses, generally leading to a preference for early IOL implantation. PMID- 15899453 TI - After-cataract in children having cataract surgery with or without anterior vitrectomy implanted with a single-piece AcrySof IOL. AB - PURPOSE: To evaluate whether cataract surgery in children should be performed with anterior vitrectomy and to examine the properties of the AcrySof SA30AL intraocular lens (IOL) in the pediatric eye. SETTING: Filatov Institute, Odessa, Ukraine. METHODS: Cataract surgery was performed in 66 children aged 3 to 15 years. They were randomized to surgery with or without anterior vitrectomy. All eyes were implanted with the single-piece AcrySof SA30AL IOL (Alcon). During the study, the patients who needed surgery for after-cataract had a second surgical procedure. Two years after surgery, the surgical method was evaluated using exact logistic regression. Also, the Evaluation of Posterior Capsule Opacification (EPCO) score was compared between the patients who had surgery for after-cataract and the patients who did not need this. The presence of posterior synechias and centration of the IOL were assessed. RESULTS: Children in the younger age group ( -6.01 D). Preoperative and postoperative spherical equivalent (SE), best corrected visual acuity (BCVA), and uncorrected visual acuity (UCVA) were measured and analyzed within each group. Patients had at least 2 follow-up visits, with the second visit occurring between 1 and 9 months postoperatively. RESULTS: Preoperatively, the mean SE in Group A was -3.93 D +/- 1.53 (SD) and in Group B, -8.49 +/- 1.70 D. At the final visit, the SE in Group A decreased to 0.29 +/- 0.55 D and in Group B, -1.09 +/- 0.87 D. The UCVA in Group A at the final visit was 20/20 or better in 43% of eyes and 20/40 or better in 100% of eyes. In Group B, the UCVA was 20/20 or better in 31% of eyes and 20/40 or better in 75% of eyes. In all eyes, the BCVA improved by 1 line in 16% or remained the same in 84%. No eyes lost any lines of BCVA. The SE taken at the last visit was within +/-0.5 D in 69% of all eyes and within +/-1.0 D in 82% of all eyes. The incidence of patients needing retreatment in resident-performed LASIK surgery was 3 of 44 (6.8%). CONCLUSIONS: Laser in situ keratomileusis performed by residents is a safe and effective procedure for correction of refractive error. Postoperative UCVA, refractive error, retreatment rate, and loss of BCVA were consistent with published results from nonresident surgeons. These results also exceeded the U.S. Food and Drug Administration requirements for LASIK surgery. PMID- 15899456 TI - Laser in situ keratomileusis for primary hyperopia. AB - PURPOSE: To evaluate the efficacy, predictability, stability, and safety of hyperopic laser in situ keratomileusis (H-LASIK) over a 24-month period and analyze topographic changes after H-LASIK to assess topographic pseudokeratectasia (TPKE) following H-LASIK. SETTING: Eye Institute of Utah, Salt Lake City, Utah, USA. METHODS: This prospective study included 139 eyes of 77 patients having H-LASIK for primary hyperopia. The mean follow-up was 15.6 months +/- 7.6 (SD) (range 6 to 48 months). One hundred twenty-two eyes (88%) were followed at 1 year and 36 eyes (26%) at 24 months. Topographic pseudokeratectasia was defined as 1 or more positive keratoconus screening findings in an eye with topographic central or inferior steepening detected by the Topography Modeling System but without corneal thinning or progressive change. RESULTS: The mean spherical equivalent manifest refraction was +2.39 +/- 0.99 diopter (D) preoperatively and -0.05 +/- 0.61 D at the last visit. Ninety-one percent of eyes were within +/-1.00 D of emmetropia and 71% of the eyes were within +/-0.50 D. Uncorrected visual acuity of 20/20 or better was present in 42%, 20/25 in 63%, and 20/40 or better in 93% of eyes. Loss of 2 lines of BSCVA occurred in 2 eyes (1.4%). In 1 eye, ischemic optic neuropathy occurred, and in another, choroidal neovascularization developed postoperatively. Topographic pseudokeratectasia was detected in 28% to 56% of eyes postoperatively. No significant difference between postoperative visual and refractive outcome, regression, or irregularity was found between the eyes with or without TPKE. CONCLUSION: Hyperopic LASIK appears to be an effective, predictable, and safe procedure to correct low to moderate primary hyperopia. Topographic pseudokeratectasia, which was observed after H LASIK with a keratoconus-like topographic pattern in otherwise normal eyes, may represent a relatively static condition. PMID- 15899457 TI - Utility assessment among cataract surgery patients. AB - PURPOSE: To evaluate the utility values (time trade-off, standard gamble for death, standard gamble for blindness) of patients scheduled for cataract surgery. SETTING: Tertiary eye hospital in Singapore. METHODS: Questions on time trade off, standard gamble for death, standard gamble for blindness utilities, and visual function from an adapted version of the VF-14 were asked in a clinic interview of 217 Chinese patients. RESULTS: The mean utility time trade-off of cataract surgery patients was 0.77 (95% confidence interval [CI] 0.76, 0.78), mean standard gamble for death was 0.77 (95% CI 0.77, 0.78), and mean standard gamble for blindness was 0.76 (95% CI 0.75, 0.77). Adults with lower VF-14 scores had lower utilities, although there was no relationship between visual acuity and utilities. Utilities were also lower in patients who reported difficulty seeing in bright light. CONCLUSIONS: The utilities of cataract patients scheduled for surgery were comparable to those in patients with other chronic ocular diseases. Cataract surgery patients with lower utilities tended to have worse visual function and greater difficulty seeing in bright light. PMID- 15899458 TI - Comparison of dyes for cataract surgery. Part 1: cytotoxicity to corneal endothelial cells in a rabbit model. AB - PURPOSE: To investigate the corneal endothelial cytotoxicity of dyes for capsule staining in cataract surgery. SETTING: Department of Ophthalmology and Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan. METHODS: Cultured corneal endothelial cells of New Zealand white rabbits were exposed for 1 minute to 1 of the following dyes (various concentrations): indocyanine green (ICG), methylene blue (MB), gentian violet (GV), trypan blue (TB), and fluorescein sodium (FS). The degree of cell damage was determined by in vitro staining with TB and comparison with results in a control group. The effect of longer exposure (up to 10 minutes) to ICG 0.25% was also investigated. Structural changes in corneal endothelial cells after dye exposure were evaluated by light microscopy and transmission electron microscopy (TEM). RESULTS: Indocyanine green 0.25%, MB 0.20%, GV 0.01%, TB 0.40%, and FS 10% did not induce significant damage to corneal endothelial cells. Significant cytotoxicity was observed with the following or higher dye concentrations: ICG 0.50%, MB 0.50%, and GV 0.10%. Exposure to ICG 0.25% for 1 to 10 minutes showed a trend toward cytotoxicity after 10 minutes. On TEM, corneal endothelial cells that had been exposed to ICG 0.50% showed remarkable organelle swelling and disruption, electron-dense granules, and cell lysis. CONCLUSION: One minute of exposure to ICG 0.25%, MB 0.20%, GV 0.01%, TB 0.40%, and FS 10% appeared to be safe as determined by no cytotoxic effects on rabbit corneal endothelial cells in culture. PMID- 15899459 TI - Comparison of dyes for cataract surgery. Part 2: efficacy of capsule staining in a rabbit model. AB - PURPOSE: To investigate the efficacy of various dyes for anterior capsule staining to facilitate capsulorhexis during cataract surgery. SETTING: Department of Ophthalmology and Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan. METHODS: Various concentrations of indocyanine green (ICG), methylene blue (MB), gentian violet (GV), trypan blue (TB), and fluorescein sodium (FS) were used to stain rabbit lenses in vitro. After 1 minute of exposure, the staining of each lens was evaluated using a semiquantitative scoring system in which 0 represented no staining/no contrast between the capsule and cortex and 4 represented excellent staining/remarkable contrast between the stained capsule and unstained cortex. The lowest concentration of dye with a score of 4 was considered the lowest effective concentration for capsulorhexis. RESULTS: Based on our scoring system, the lowest effective concentrations for capsulorhexis were ICG 0.25%, MB 0.10%, GV 0.01%, TB 0.10%, and FS 1.25%. The lowest effective concentrations of the 5 dyes provided comparable contrast. Increased concentrations of dye decreased contrast between the capsule and the cortex and were considered less effective for capsulorhexis. CONCLUSION: Any of the following concentrations of dyes can be used for optimal enhancement of anterior capsule visibility for capsulorhexis: ICG 0.25%, MB 0.10%, GV 0.01%, TB 0.10%, and FS 1.25%. PMID- 15899460 TI - Posterior capsule opacification in rabbit eyes implanted with 1-piece and 3-piece hydrophobic acrylic intraocular lenses. AB - PURPOSE: To evaluate the outcome of posterior capsule opacification (PCO) after implantation in rabbit eyes of currently available 3-piece and 1-piece hydrophobic acrylic intraocular lenses (IOLs) with square optic edges. SETTING: John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: The 3-piece designs evaluated were the AR40e (Advanced Medical Optics Inc.) and the MA60AC (Alcon, Inc.); the 1-piece designs were the SA60AT and the SA30AT (Alcon, Inc.). Nine lenses of each type were implanted in a randomized manner by the same surgeon in 18 Dutch Belted pigmented rabbits. After a follow-up of 3 weeks, the rabbits were killed and analyses of the enucleated eyes were performed from the posterior or Miyake-Apple view. The intensity of central PCO, peripheral PCO, and Soemmering's ring formation was scored from 0 to 4. The area of Soemmering's ring formation was also scored from 0 to 4 based on the number of quadrants involved. Other parameters analyzed were capsulorhexis coverage of the IOL anterior surface, IOL centration, fixation, and presence of striae. Results from the posterior view were complemented by histopathologic evaluation of the eyes. RESULTS: No statistically significant difference was found between the 4 groups of IOLs in the parameters analyzed from the posterior view. When cell ingrowth occurred with the 1-piece designs, causing peripheral and central PCO formation, it was more likely to start at the optic-haptic junctions, as observed during the clinical follow-up with slitlamp examination and confirmed by gross and histopathologic analyses of the enucleated eyes. CONCLUSIONS: The square, truncated optic edge is the most important IOL design feature for PCO prevention. The optic-haptic junctions of the 1-piece designs appear to be sites where the barrier effect of the truncated optic edge is less effective. PMID- 15899461 TI - Thermal comparison of the AdvanTec Legacy, Sovereign WhiteStar, and Millennium phacoemulsification systems. AB - PURPOSE: To determine the operating temperature of ultrasonic tips used with the Alcon AdvanTec Legacy, AMO Sovereign WhiteStar, and Bausch & Lomb Millennium phacoemulsification systems. SETTING: Mackool Eye Institute, Astoria, New York, New York, USA. METHODS: Thermal imaging of ultrasonic tips (Legacy, WhiteStar, and Millennium) was performed in air and in human cadaver eyes using a duty cycle of 33% (WhiteStar) and 50% (Millennium and Legacy). In vitro temperatures were measured with the tip centered in the incision and intentionally decentered against the side of the incision. The stroke length of each instrument was also measured, and the operating frequency of the Legacy was evaluated with the addition of a tangential load. RESULTS: Open air and in vitro testing demonstrated that tip temperatures with the Legacy were consistently the lowest. Temperatures measured with the WhiteStar and Millennium systems were higher and generally similar to each other. At identical console power settings, the stroke length of the WhiteStar and Millennium tips was longer than that of the Legacy. The frequency of the Legacy handpiece did not change significantly (less than 200 Hz) under conditions of tangential tip loading. CONCLUSIONS: At identical console power settings and similar console duty cycles, the temperature elevation of ultrasonic tips was least for the Legacy and greater for the WhiteStar and Millennium under all conditions. The causes of these findings appear to be the longer stroke length of the WhiteStar and Millennium and the underestimation of the duty cycle with the WhiteStar. PMID- 15899462 TI - Intraocular lens-capsular bag imaging with ultrahigh-resolution optical coherence tomography Pseudophakic human autopsy eyes. AB - PURPOSE: To compare in vitro ultrahigh-resolution optical coherence tomography (UHR OCT) cross-sectional images of the pseudophakic human autopsy eye with histology to evaluate the potential of this imaging technique for enhanced visualization of the anterior segment, especially the capsular bag, intraocular lens (IOL), and posterior capsule opacification (PCO) formation after cataract surgery. SETTING: Department of Medical Physics and Department of Ophthalmology, University of Vienna, Vienna, Austria, and Department of Oral and Maxillofacial Surgery Institute of Dentistry, University of Turku, Turku, Finland. METHODS: Ultrahigh-resolution OCT images were acquired from 7 pseudophakic human autopsy eyes using 1.4 microm axial x 3.0 microm transverse resolution. The axial resolution with UHR OCT is 1.4 microm compared to 10.0 microm with the commercially available OCT. Plastic-embedded histologic sections were obtained in precise alignment with the OCT tomograms. RESULTS: Ultrahigh-resolution OCT cross sectional tomograms corresponded to the histologic sections. With the wavelength used (800 nm), the anterior and posterior capsules, area of lens epithelial cell growth and extracellular matrix proliferation, and IOL could be clearly visualized. The extent of capsular bag adhesion to the IOL could be detected, as well as the amount of PCO formation. CONCLUSIONS: The improved resolution makes UHR OCT a powerful tool in anterior segment imaging and evaluation of the capacity of IOL materials and models to induce capsular bag adhesion. Ultrahigh resolution OCT may also help in determining the area of origin of PCO after cataract surgery. PMID- 15899463 TI - Cataracts in children. AB - Bilateral congenital cataract is the most common cause of treatable childhood blindness. Nuclear cataract is usually present at birth and is nonprogressive, whereas lamellar cataract usually develops later and is progressive. Surgery must be performed promptly in cases with dense congenital cataract; if nystagmus has developed, the amblyopia is irreversible. A treatment regimen based on surgery within 2 months of birth combined with prompt optical correction of the aphakia and aggressive occlusion therapy with frequent follow-up has been successful in unilateral and bilateral cases. Both anterior and posterior capsulorhexes are performed in most children. Intraocular lens implantation can be performed safely in children older than 1 year. Anterior dry vitrectomy is recommended in preschool children to avoid after-cataract. Opacification of the visual axis is the most common complication of cataract surgery in children. Secondary glaucoma is the most sight-threatening complication and is common if surgery is performed early. Life-long follow-up is essential in these cases. PMID- 15899464 TI - Negative dysphotopsia after intraocular lens implantation irrespective of design and material. AB - We report 3 cases of negative dysphotopsia in patients who had implantation of 1 intraocular lens (IOL) type in 1 eye and another IOL type in the fellow eye. A male patient (aged 58 years) received an SA60AT AcrySof hydrophobic acrylic IOL (Alcon) in his left eye (OS). He noticed a temporal dark shadow. Three months later, he received a Tecnis Z9001 silicone aspheric optic IOL (Pfizer) in his right eye (OD). He noticed a temporal dark shadow OD almost identical to the shadow OS. Another male patient (aged 58 years) received an AcrySof MA60AC IOL (Alcon) OS and subsequently a PhacoFlex SI-40NB silicone IOL (AMO) OD. He experienced temporal dark shadows in each eye. A female patient (aged 64 years) received an SA60AT IOL OD and subsequently a Sensar hydrophobic acrylic AR40e (AMO) OS, noticing dark shadows in both eyes. Although all 3 patients tolerated the shadows well, they were interested in their etiology and wanted reassurance that the symptoms would not progress. PMID- 15899465 TI - Negative dysphotopsia associated with implantation of the Z9000 intraocular lens. AB - Uneventful small-incision cataract surgery was performed in a 70-year-old man and a 62-year-old woman with implantation of a Tecnis Z9000 intraocular lens (IOL) (Pharmacia). No ocular pathology was present in the patients preoperatively. Unsolicited complaints of negative dysphotopsia were made postoperatively, and the symptoms have persisted for more than 1 year in both patients. Symptomatic negative dysphotopsia may be seen in association with the Z9000 IOL. PMID- 15899466 TI - Use of preservative-free lidocaine for cataract surgery in a patient allergic to "caines". AB - Although many patients have been labeled allergic to local anesthetics (LAs), true allergic reactions to LAs are rare. An 81-year-old woman with a history of procaine (Novocaine) allergy presented for cataract surgery. Skin testing showed sensitivity to amide and ester LAs. Further testing with preservative-free lidocaine was negative, suggesting the patient was allergic to ester LAs and preservatives found in amide anesthetic preparations. Cataract extraction was subsequently and uneventfully performed in both eyes with topical anesthesia using preservative-free lidocaine. PMID- 15899467 TI - Complicated removal of corneal foreign bodies 18 months after laser in situ keratomileusis. AB - We present a 39-year-old young engineer who developed flap complications after removal of corneal foreign bodies 18 months after successful laser in situ keratomileusis. The right eye had a flap scar and debris in the interface, and the left eye had a central rust ring and debris in the interface. Attempted removal of the flap in the left eye lifted off a large area around the rust ring, and the ring was left in place. One week after treatment with topical antibiotics, the eye was white and the rust ring had begun to disappear. The ring was successfully removed without complications the following week. PMID- 15899468 TI - Imaging interface fluid after laser in situ keratomileusis with corneal optical coherence tomography. AB - A 41-year-old myopic patient who had laser in situ keratomileusis 6 months earlier was treated for a complete retinal detachment (RD) with proliferative vitreoretinopathy. Surgical treatment consisted of an encircling band, pars plana vitrectomy, and silicone oil filling. Postoperatively, the patient developed marked corneal edema with no increase in intraocular pressure (IOP) as measured by applanation tonometry. Interface fluid was confirmed by corneal optical coherence tomography. Quantification of the corneal structures revealed that corneal edema was in the residual posterior stroma predominantly. The epithelial and flap thickness did not change significantly. The case demonstrated that after vitreoretinal surgery for RD repair, transient corneal endothelial cell dysfunction developed, causing marked edema of the posterior corneal stroma and interface fluid accumulation. However, an increase in IOP cannot be excluded. PMID- 15899469 TI - Laser in situ keratomileusis induced corneal perforation and recurrent corneal epithelial ingrowth. AB - We report a rare complication of recurrent corneal epithelial cell ingrowth after corneal perforation induced by laser in situ keratomileusis. A corneal flap was made uneventfully with an automatic SCMD microkeratome (Howard Instruments, Inc.). During laser ablation, instead of random burning over the stromal bed, repeated ablation at a single location occurred because of a machine malfunction, resulting in corneal perforation. The flap was replaced and sutured to prevent leakage of aqueous humor. As the cornea healed, the patient's vision was compromised by scar formation, high irregular astigmatism, and epithelial ingrowth. The flap was reopened, and the epithelial cells were removed by thoroughly scraping the flap and stromal bed. However, corneal epithelial ingrowth recurred within a week. The recurrent and progressive epithelial ingrowth further decreased vision, and penetrating keratoplasty was recommended. PMID- 15899470 TI - Cataract surgery in Stevens-Johnson syndrome. AB - Cataract surgery can be safely performed in patients with Stevens-Johnson syndrome (SJS) with visually significant cataracts. Maintaining ocular surface integrity improves the prognosis of cataract surgery in this high-risk population. We present the outcome of cataract surgery in a retrospective chart review of 3 eyes in 2 patients with SJS. Uneventful extracapsular cataract extraction with intraocular lens implantation was performed. Follow-up ranged from 3 to 24 months. The visual acuity and ocular surface integrity were reviewed. The preoperative visual acuity was counting fingers at 1 meter in all 3 eyes. The best corrected visual acuity (BCVA) improved to 20/40 in 2 eyes and 20/50 in 1 eye postoperatively. A drop in BCVA ranging from 20/100 to 20/200 was noted during follow-up. PMID- 15899471 TI - Understanding spatial concentrations of road accidents using frequent item sets. AB - This paper aims at understanding why road accidents tend to cluster in specific road segments. More particularly, it aims at analyzing which are the characteristics of the accidents occurring in "black" zones compared to those scattered all over the road. A technique of frequent item sets (data mining) is applied for automatically identifying accident circumstances that frequently occur together, for accidents located in and outside "black" zones. A Belgian periurban region is used as case study. Results show that accidents occurring in "black" zones are characterized by left-turns at signalized intersections, collisions with pedestrians, loss control of the vehicle (run-off-roadway) and rainy weather conditions. Accidents occurring outside "black" zones (scattered in space) are characterized by left turns on intersections with traffic signs, head on collisions and drunken road user(s). Furthermore, parallel collisions and accidents on highways or roads with separated lanes, occurring at night or during the weekend are frequently occurring accident patterns for all accident locations. These exploratory results show the potentiality of the frequent item set method in addition to more classical statistical techniques, but also suggest that there is no unique countermeasure for reducing the number of accidents. PMID- 15899472 TI - Identification of a drug target motif: an anti-tumor drug NK109 interacts with a PNxxxxP. AB - The synthetic compound NK109 shows anti-tumor effects against a number of human cancer cell lines. The mechanism of action is thought to involve the inhibition of DNA topoisomerase II. However, NK109 also exhibits potent anti-tumor activities against doxorubicin-, cisplatin- and etoposide-resistant human cell lines. This paper describes target validation of NK109 using biotinylated NK109 and a T7 phage library screening procedure. Phage particles displaying an affinity for NK109 were isolated and the DNA sequence determined. The amino acid sequences of selected peptides, and the results of mutation experiments by alanine scanning, confirmed that the binding target motif of NK109 is PNxxxxP. In silico analysis of the interaction between NK109 and the peptide, by docking simulation and molecular dynamics, supported this conclusion. The PNxxxxP motif exists in the C2 domain of protein kinase Calpha. NK109 was confirmed to bind the C2 domain from surface plasmon resonance analysis. Furthermore, NK109 moderately inhibited protein kinase C activity in vitro. Our results show that the anti tumor activity of NK109 stems from interactions with multiple protein targets. PMID- 15899473 TI - Redox regulation: a new challenge for pharmacology. AB - Redox signaling is evolving as a new field of biochemical and pharmacological research. Unlike oxidative stress which is characterized by a macroscopic shift in cellular redox potentials and usually accompanied by oxygen radical induced damage, redox regulation involves subtle and more chemically defined oxidations of short duration. Most important is the reductive component as a necessary part of a reversible regulatory process. Examples of redox regulation occur during early stages of the immune response, in hypoxia or in endothelial dysfunction. Persistent oxidative events together with a decline in the cellular reduction potential lead to oxidative stress as is seen in the pathophysiology of sepsis, reperfusion damage, atherosclerosis and diabetes. Oxidative signals involve superoxide and nitric oxide as the main players which form a system of oxidizing, nitrating or nitrosating species leading to posttranslational modifications of proteins. Modern techniques of immunohistochemistry and mass spectrometry allow a correlation of protein modification, e.g., disulfide, S-oxide, S-nitroso or nitrotyrosine formation, with enzyme activities and cellular responses. In this commentary, examples of the control of prostanoid synthesis by the NO/O2- system are described. Redox regulation represents an interesting challenge for the development of drugs that modulate the oxidative trigger mechanisms or enforce the reductive pathways. PMID- 15899474 TI - Z-analysis: a new approach to analyze stimulation curves with intrinsic basal stimulation. AB - In the study of receptor biology it is of considerable importance to describe the stimulatory properties of an agonist according to mathematically defined models. However, the presently used models are insufficient if the experimental preparation contains an intrinsic basal stimulation. We have developed a novel approach, tentatively named Z-analysis. In this approach, the concentration of endogenous agonist is calculated by extending the stimulation curve to zero effect. The concentration of endogenous agonist is then combined with the concentration of added agonist to estimate the true EC(50) value. We developed a new model, the Z-model, specifically for this purpose, but in addition, we describe how Z-analysis can be applied to the traditional E(0)-model. Models were applied to computer-generated curves with different Hill coefficients, using iterative curve fitting procedures. In addition to applying the models to ideal cases, we also used Monte Carlo-simulated data. Specific transformations were used to enable comparisons between parameters determined from these models. Both models were able to provide estimates of all eight parameters analyzed, both using ideal data and on Monte Carlo-simulated data. The Z-model was found to provide better estimates of the concentration of endogenous agonist, the EC(50) values, and the Hill value, in curves with Hill coefficient deviating from one. In conclusion, Z-analysis was suitable both to determine the concentration of endogenous agonists and to determine true EC(50) values. We found several advantages with the Z-model compared to traditional E(0)-model for analysis of stimulation curves that contain basic intrinsic stimulation. PMID- 15899475 TI - Human epidermal cell protein responses to arsenite treatment in culture. AB - Study of the responses of target cells in culture is anticipated to help understand the mechanisms by which inorganic arsenic causes pathological effects in vivo. Treatment of human epidermal cells with arsenic has been shown to produce a myriad of changes in gene transcription. Present work focused on finding the extent of arsenite-induced changes in the protein pattern and whether global effects on protein sulfhydryls were evident. First, examining the profile of protein expression by two-dimensional gel electrophoresis indicated that approximately 40% of the 300 distinct protein spots that were monitored changed by at least two-fold in amount all through a 9-day exposure period. Second, examining soluble extracts of the treated cells by Activated Thiol Sepharose column chromatography gave little indication of change in the overall protein thiol content. Finally, among the 10 proteins identified that showed prominent changes in amount as a result of treatment for 1 or 4 days, enzymes of the glycolytic pathway were seen to be substantially elevated as a result of treatment, suggesting decreased utilization by the cells of oxidative phosphorylation. Since these changes were more conspicuous at the protein level than in previous transcriptional studies, the results emphasize the importance of proteomic analysis to complement transcriptional analysis of cell responses to perturbation by arsenic. PMID- 15899476 TI - Racemic and optically active 2-methoxy-4-oxatetradecanoic acids: novel synthetic fatty acids with selective antifungal properties. AB - The unprecedented (+/-)-2-methoxy-4-oxatetradecanoic acid and the optically pure (S)-2-methoxy-4-oxatetradecanoic acid were synthesized in six steps and in 11-14% overall yields starting with either 1,2-O-isopropylidene-rac-glycerol or 1,2-O isopropylidene-(S)-glycerol. The key step in the synthesis was the selective monosilylation of a dibutylstannylene intermediate. The title compounds displayed selective fungitoxicity in the range of 0.08-0.22 mM against Cryptococcus neoformans ATCC 66031 and Aspergillus niger ATCC 16404, but no significant activity against C. albicans ATCC 14053 and ATCC 60193 (>2.6 mM). Albeit being good substrates for N-myristoyltransferases (NMTs), the racemic and the S enantiomer of the oxygenated 2-methoxylated compounds showed no significant difference in antifungal activity. This finding suggests an alternative mechanism of fungitoxicity other than NMT inhibition. PMID- 15899477 TI - Novel indices of ventricular repolarization to screen post myocardial infarction patients. AB - We propose novel indices of ventricular repolarization intervals, the JTp/JT, Tpe/JTp and Tpe/JT ratios. These indices have been compared with the duration of the ventricular repolarization intervals and other ratios in 17 normal subjects and 17 patients with old myocardial infarction. In the intervals and other ratios, the best separation between groups is obtained with the Tpe/QTp and Tpe/QT ratios with 94% sensitivity and 82% specificity, the proposed ratios increased sensitivity to 100% and specificity to 94%. These indices should be further tested to determine their usefulness in discriminating between OMI patients with and without susceptibility to ventricular arrhythmias. PMID- 15899478 TI - 3,4-Methylenedioxymethamphetamine and naloxone in rat rotational behaviour and open field. AB - It has recently been shown that 3,4-Methylenedioxymethamphetamine (MDMA) has an anti-parkinsonian effect in rodent models of Parkinson's disease. The mechanism of this anti-parkinsonian action is unknown. Opioids have been suggested to play a role in MDMA-induced behaviour. We therefore investigated MDMA and naloxone in the rat rotational behavioural model. Male Sprague-Dawley rats were lesioned unilaterally with 6-hydroxydopamine at the medial forebrain bundle. Administration of R/S-MDMA (5 mg/kg, s.c.) produced ipsilateral rotations. Naloxone (2, 5, 10 mg/kg, s.c.) did not produce rotations on its own but reduced the number of MDMA-induced ipsilateral rotations. This effect was not dose dependent. In contrast to reports on mice, in unlesioned animals, naloxone (10 mg/kg, s.c.) did not block MDMA (5 mg/kg, s.c.)-induced hyperactivity in an open field in our experiment. It is concluded that endogenous opioids play a role in MDMA's action in the rat rotational behavioural model. PMID- 15899479 TI - Dual impact of a nitric oxide donor, GEA 3175, in human pulmonary smooth muscle. AB - Nitric oxide (NO) donors could constitute an alternative to inhaled NO as treatment in some patients with pulmonary hypertension. Therefore, the present study investigated the relaxation mechanisms of a novel NO donor, 3-(3-chloro-2 methylphenyl)-5-[[4-methylphenyl)sulphonyl]amino]-)hydroxide (GEA 3175) in segments of human pulmonary arteries and bronchioles, which were mounted in microvascular myographs. GEA 3175 induced concentration-dependent relaxations and was more potent in pulmonary arteries than in bronchioles. A blocker of soluble guanylyl cyclase, 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ), and iberiotoxin, a blocker of large-conductance calcium-activated K channels, both reduced relaxations induced by GEA 3175 in pulmonary arteries and bronchioles. Combining of ODQ and iberiotoxin did not produce additional inhibition. GEA 3175 relaxation is mediated through guanylyl cyclase-dependent mechanisms followed by activation of large-conductance calcium-activated K(+) channels. The dilatation of both pulmonary small arteries and airways by GEA 3175 seems advantageous, if it is considered administered as inhalation therapy for pulmonary hypertension. PMID- 15899480 TI - Inhibition by agmatine on morphine-induced conditioned place preference in rats. AB - Our previous studies demonstrated the ability of exogenous agmatine to inhibit tolerance to and physical dependence on morphine in mice, rats and monkeys. The present study further evaluated the effect of agmatine on the psychological dependence induced by morphine in conditioned place preference assay. Agmatine (0.75-20 mg/kg, s.c.) co-administered with morphine during the conditioning sessions completely abolished the acquisition of morphine-induced conditioned place preference in rats, which was associated with activation of imidazoline receptors. Agmatine (0.75-10 mg/kg, s.c.) administered on the test day inhibited the expression of the place preference. After 30 days of extinction of conditioned place preference, agmatine 2.5 and 40 mg/kg inhibited the priming effect of morphine 0.5 mg/kg on the place preference. Furthermore, agmatine inhibited the increased expression of FosB in the nucleus accumbens caused by chronic morphine. All these results suggest that agmatine could inhibit morphine induced psychological dependence and relapses by affecting the expression of transcription factor FosB. PMID- 15899481 TI - Dose response effects of lithium chloride on conditioned place aversions and locomotor activity in rats. AB - The present study examined the multi-variable locomotor activity effects of lithium chloride (LiCl) treatment in male rats. Of interest was a determination of which variables might show a dose-response relationship in LiCl-induced conditioned place aversions. Automated open-fields were partitioned into two chambers distinct in tactile and visual cues. A control group [n=8] received saline (NaCl; 0.15 M) paired with both chambers while three LiCl groups (0.15 M; 32 mg/kg [n=7], 95 mg/kg [n=7], 127 mg/kg [n=7]) received LiCl paired with the normally preferred chamber and saline paired with the non-preferred chamber. During extinction trials, rats were allowed to choose between the two chambers to provide an index of conditioned place aversions. Locomotor activity and its distribution within the chambers were also assessed during both conditioning and extinction trials. Dose-dependent decreases occurred in all measures of locomotor activity following LiCl administration during conditioning. During extinction trials, place aversions developed in animals conditioned with LiCl. LiCl-treated rats spent significantly less time in the LiCl-paired chamber relative to controls but not in a dose-dependent manner. Animals that had been conditioned with 95 or 127 but not 32 mg/kg LiCl, displayed significantly more vertical activity in the LiCl-paired chamber than controls during extinction trials. These findings indicate that, in addition to producing dose-dependent unconditioned effects on locomotor activity, LiCl also produces dose-dependent conditioned effects on vertical activity. These conditioned rearing response effects provide a valid measure of the conditioned avoidance response that provides evidence for dose-dependent LiCl-induced conditioned place aversions. PMID- 15899482 TI - Rheumatoid arthritis and anti-TNF-alpha therapy. PMID- 15899483 TI - Identification of mildly oxidized low-density lipoprotein (electronegative LDL) and its auto-antibodies IgG in children and adolescents hypercholesterolemic offsprings. AB - BACKGROUND: Oxidative modification of low-density lipoproteins (LDL) is an essential step in atherogenesis, generating minimally oxidized LDL, also called electronegative LDL [LDL(-)], which has chemotactic, cytotoxic and immunogenic properties. METHODS AND RESULTS: Serum LDL(-) and anti-LDL(-) auto-antibodies (IgG) were evaluated in 28 children and adolescents with familial hypercholesterolemia (FH) antecedents, with or without early coronary artery disease in first-degree relatives (eCAD), hypercholesterolemic (hc) or normocholesterolemic (nc) versus a control group of normocholesterolemic children without pathologic antecedents (C). ELISA method was used for detection of LDL(-) and anti-LDL(-) IgG. LDL(-) serum levels did not differ among the four groups (FH eCAD-hc 41.4 +/- 24.9 microg/dl; FH-hc 38.3 +/- 11.2 microg/dl; FH-nc 47.3 +/- 17.0 microg/dl and C 44.2 +/- 28.8 microg/dl, p = 0.659). However, IgG anti-LDL( ) auto-antibodies were significantly higher in the control group in comparison to the FH groups with or without eCAD, independent of hypercholesterolemia or normocholesterolemia (FH-eCAD-hc 0.825 +/- 0.289 microg/dl; FH-hc 0.667 +/- 0.307 microg/dl; FH-nc 0.763 +/- 0.204 microg/dl and C 1.105 +/- 0.233 microg/dl, p = 0.006). When the auto-antibodies of groups with FH, with or without eCAD and with or without hypercholesterolemia were compared, no differences were found (p = 0.509). CONCLUSION: These results showed that FH and/or eCAD children and adolescents have lower titers of auto-antibodies anti-LDL(-) than children from normal families, independent of serum LDL-cholesterol or serum LDL(-). PMID- 15899484 TI - Genotype of the mutant LDL receptor allele is associated with LDL particle size heterogeneity in familial hypercholesterolemia. AB - Small, dense LDL particles have been associated with an increased risk of coronary artery disease. In order to assess the potential contribution of the genotype of the LDL receptor to LDL particle size heterogeneity in familial hypercholesterolemia (FH), we examined the electrophoretic characteristics of LDL particles in a large cohort of FH heterozygotes and controls. A total of 259 FH heterozygotes and 208 controls participated in the study. FH subjects were carriers of one of the nine French Canadian mutations in the LDL receptor gene. LDL particles were characterized by polyacrylamide gradient gel electrophoresis following a 6-week lipid-lowering drug-free baseline period. LDL-peak particle diameter (LDL-PPD), representing the most abundant LDL particle subpopulation, was significantly smaller in FH heterozygotes carrying a negative-receptor mutation than in subjects carrying a defective-receptor mutation (negative receptor = 257.3 +/- 4.1 A versus defective-receptor = 259.0 +/- 4.3 A, p = 0.0006). No significant difference in plasma CETP concentrations was found between these two genotypic groups. Moreover, compared with controls having low triglyceride levels, negative-receptor subjects with high triglyceride levels had a relative risk of 19.6 (p < 0.0001) of having small, dense LDL particles while this risk was not significantly increased among defective-receptor subjects. Multivariate analysis showed that the LDL receptor status accounted for 5.7% of the variance in the LDL-PPD after adjustment for covariates. These results suggest that the genotype of the mutant LDL receptor allele was independently associated with variations in LDL-PPD and could partly explain why negative receptor FH heterozygotes may be at greater risk of cardiovascular disease than defective-receptor FH subjects. PMID- 15899485 TI - Paraoxonase-1 activity modulates endothelial function in patients with peripheral arterial disease. AB - Human serum paraoxonase-1 (PON1) is thought to play a role in the favorable vascular effects of high-density lipoproteins, mainly through a reduction in low density lipoprotein oxidation. Endothelial dysfunction, characterized by an impaired capacity of the arteries to dilate in response to a number of stimuli, represents the earliest stage of atherosclerosis. We performed the present study in 37 patients with peripheral arterial disease, with the aim of investigating the influence of PON1 Q192R polymorphism and activity on peripheral endothelial function, evaluated as brachial-artery flow-mediated vasodilation (FMV). Patients with the R allele (QR or RR genotype, n=19) had significantly higher PON1 activity [408 U/mL (309-456) versus 180 U/mL (141-243), p<0.001] and greater brachial FMV (5.7+/-3.9% versus 3.0+/-2.8%, p<0.001) than those with Q allele (QQ genotype, n=18). In the whole population, PON1 activity showed a direct relation to brachial FMV (r=0.46, p=0.004). In a multivariate linear regression analysis, the only independent predictors of brachial FMV were PON1 activity (beta=0.40, p=0.008), brachial-artery diameter (beta=-0.39, p=0.01) and male sex (beta=-0.27, p=0.04). These finding support the importance of PON1 activity as a modulating factor of the endothelial function. PMID- 15899486 TI - Phenotypic modifications of vascular smooth muscle cells could be responsible for vascular hyporeactivity to contracting agent in mechanically injured rat carotid artery. AB - Vascular smooth muscle cells (VSMCs) that accumulate in neointima after angioplastic injury show different phenotypic characteristics from those of medial layer and an impaired reactivity to contracting agents. The aim of the study was to correlate the vascular hyporesponsiveness to the changes in intracellular calcium concentration [Ca(2+)](i) and the expression of proteins necessary for its utilization in mechanically injured rat carotid arteries (IC) at 14 and 28 days after angioplastic balloon. IC showed a significant reduction (P<0.01) to PE- or KCl-induced contraction as compared to uninjured carotid (UC). Fura-2AM-loaded VSMCs isolated from IC revealed that this hyporeactivity to PE or KCl was accompanied by the impairment of the increase in [Ca(2+)](i) induced by contracting agents in both Ca(2+)-free or -containing medium. Similar results were observed following the ryanodine challenge in VSMC. Western blot analysis showed a significant (P<0.05) reduction in myosin heavy chain (MHC) and IP(3) type III receptor expression in IC isolated at 14 days from injury compared to UC, while an improvement of these proteins expression was observed at 28 days after damage. On the other hand, in IC tissue, SERCA2 and alpha-actin expression, compared to UC was significantly higher at 14 days than at 28 days. These data indicate that vascular hyporeactivity induced by mechanical injury may be due to alterations of either [Ca(2+)](i) or contractile proteins. These modifications could be related to the changes of VSMC phenotypic characteristics, as supported by the observed modifications in MHC, SERCA2 and alpha-actin expression, proteins considered as biological markers of cellular differentiation. PMID- 15899487 TI - RANTES gene polymorphisms predict all-cause and cardiac mortality in type 2 diabetes mellitus hemodialysis patients. AB - INTRODUCTION: Patients with type 2 diabetes (DM2) and end stage renal disease (ESRD) have a dismal survival prognosis, mainly due to cardiovascular events. There is sparse data on genetic predictors. Chemokines and their receptors are important in regulating leukocyte influx and activation in atherosclerosis, and functional polymorphisms in the respective genes are associated with cardiovascular disease risk. METHODS: We enrolled 225 prevalent Caucasian DM2 patients receiving maintenance hemodialysis in 30 centres in Southern Germany (time from dialysis initiation <2.0 years) from August 1999 to January 2000 for prospective study until December 2003. The CX3CR1 T280M, and MCP-1 -2518 and RANTES -403 and -28 promoter and intronic In1.1T/C polymorphisms were assessed by real-time PCR. Primary end point was all-cause mortality (ACM). RESULTS: Patients carrying the RANTES -403A or In1.1C allele had a significantly higher ACM risk (multivariate hazard ratio for -403A, dominant model=1.81 [95% CI: 1.22-2.67], p=0.003), mainly due to cardiac events. Similar data were obtained by haplotype analysis. The other SNPs showed no effect on survival. DISCUSSION: In DM2 patients with ESRD, ACM due to cardiac events is associated with RANTES gene variants that are known to alter the expression of this chemokine important in atherosclerosis. Further study of the role of chemokine and chemokine receptor gene variation in determining vascular end points is needed. PMID- 15899488 TI - Tensile cervical facet capsule ligament mechanics: failure and subfailure responses in the rat. AB - Clinical, epidemiological, and biomechanical studies suggest the involvement of the cervical facet joint in neck pain. Mechanical studies have suggested the facet capsular ligament to be at risk for subfailure tensile injury during whiplash kinematics of the neck. Ligament mechanical properties can be altered by subfailure injury and such loading can induce cellular damage. However, at present, there is no clear understanding of the physiologic context of subfailure facet capsular ligament injury and mechanical implications for whiplash-related pain. Therefore, this study aimed to define a relationship between mechanical properties at failure and a subfailure condition associated with pain for tension in the rat cervical facet capsular ligament. Tensile failure studies of the C6/C7 rat cervical facet capsular ligament were performed using a customized vertebral distraction device. Force and displacement at failure were measured and stiffness and energy to failure were calculated. Vertebral motions and ligament deformations were tracked and maximum principal strains and their directions were calculated. Mean tensile force at failure (2.96 +/- 0.69 N) was significantly greater (p < 0.005) than force at subfailure (1.17 +/- 0.48 N). Mean ligament stiffness to failure was 0.75 +/- 0.27 N/mm. Maximum principal strain at failure (41.3 +/- 20.0%) was significantly higher (p = 0.003) than the corresponding subfailure value (23.1 +/- 9.3%). This study determined that failure and a subfailure painful condition were significantly different in ligament mechanics and findings provide preliminary insight into the relationship between mechanics and pain physiology for this ligament. Together with existing studies, these findings offer additional considerations for defining mechanical thresholds for painful injuries. PMID- 15899489 TI - Calodium hepaticum (syn. Capillaria hepatica) in captive rodents in a zoological garden. AB - Calodium hepaticum infection was diagnosed in the Bristol Zoo Gardens in 13 captive rodents of four species that died or were humanely killed over a 40-month period. Of these infected animals, nine were black-tailed prairie dogs (Cynomus ludovicianus), representing 45% of the members of this species examined during the study. A wild rat (Rattus norvegicus) found dead in an enclosure was also infected. To date few cases of C. hepaticum infection have been reported in the UK. The number of cases diagnosed in this urban zoo may be explained by the potentially high prevalence of infection in urban rat populations and increased risk of exposure of zoo animals kept in enclosures to which rats have access. As C. hepaticum is potentially zoonotic, members of staff in zoos should be careful to avoid soil-to-mouth contact, particularly in prairie dog enclosures. PMID- 15899490 TI - Changes in the tibial growth plates of chickens with thiram-induced dyschondroplasia. AB - Tibial dyschondroplasia (TD) is a metabolic cartilage disease of young poultry in which endochondral bone formation is disrupted leading to the retention of a non calcified, avascular plug of cartilage in the tibial growth plate. Chicks aged 7 days were fed either a control diet or one containing thiram 100 ppm for 48 h to induce TD. Cell multiplication in the growth plate was determined thereafter with bromodeoxyuridine (BrdU) labelling, and metabolic changes by measuring alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP), and glutathione (GSH) activities. The effect on chondrocyte maturation was examined by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of gene expression. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) and DNA fragmentation were used to determine the effects of thiram on cell survival. The results showed that thiram-induced TD was not due to the multiplication of cells in the post-proliferative zones. Thiram did not affect ALP activity, which would have indicated a loss of calcification potential, but it reduced both TRAP and the glutathione concentrations, suggesting that the growth plate metabolism and remodelling functions were adversely affected. Thiram appeared to have no effect on the expression of type X collagen, transglutaminase, RUNX2, or matrix metalloproteinase-2 (MMP) genes suggesting that it did not alter the maturation potential of chondrocytes. On the contrary, the expressions of MMP-13 and vascular endothelial growth factor (VEGF) genes were "up-regulated," suggesting that thiram has pro-angiogenic activity. However, TUNEL assay showed that thiram induced endothelial cell apoptosis in the capillary vessels of the growth plates, as early as 10 days of age, when TD was not visually evident. The vascular death increased on subsequent days accompanied by massive death of chondrocytes in the transition zone of the growth plate. The induction of apoptosis in the growth plate was also demonstrated by DNA fragmentation. It was concluded that thiram induced TD not through an increase in the multiplication of chondrocytes in the transition zone and not by altering the expression of genes causing the arrest of chondrocytes in a prehypertrophic state, but by creating a metabolic dysfunction which led to the destruction of blood capillaries in the transition zone chondrocytes. PMID- 15899491 TI - Renal dysplasia with unilateral renal agenesis in a dog. AB - This report describes a renal dysplastic lesion associated with renal agenesis in a 3-year-old dog with chronic renal failure. Haematological examination revealed non-regenerative anaemia, azotaemia, increased creatinine and hyperphosphataemia. At necropsy, the right kidney and right ureter could not be identified. The left kidney was slightly enlarged, with a reduced cortico-medullary ratio. Histologically, the medulla of the left kidney had persistent mesenchyme and primitive tubules (tall pseudostratified columnar epithelium), dilated collecting ducts lined by flattened epithelium, and adenomatoid proliferation of cuboidal epithelium; fetal or immature glomeruli could not be identified. To our knowledge, this is the first report of a renal dysplastic lesion with unilateral agenesis in animals. PMID- 15899492 TI - Expression of proinflammatory cytokines by hepatic macrophages in acute classical swine fever. AB - Fourteen pigs were inoculated with the 'Alfort 187' strain of classical swine fever (CSF) virus and killed in pairs at 2, 4, 7, 9, 11, 14 or 17 days post inoculation for histopathological, ultrastructural and immunohistochemical examination. For the latter method, the antibodies used were those against viral antigen Gp55, porcine myeloid marker SWC3, IL-1alpha, IL-6, TNF-alpha and Factor VIII-related antigen. Activation and increase in the number of hepatic macrophages was observed following viral detection in liver, as well as an increase in IL-1alpha and IL-6 production, mainly by Kupffer cells. Maximum detection of viral antigen was observed in the middle stage of the experiment coinciding with overexpression of the three cytokines studied, with IL-6 production by interstitial macrophages prominent at the end. Additionally, the labelling of platelets for Factor VIII-related antigen and the ultrastructural study of the sinusoids revealed activation and aggregation of thrombocytes close to Kupffer cells at the beginning of the infection. The liver seems to play a prominent role in the origin of the thrombocytopenia that occurs in CSF and contributes to the overexpression of proinflammatory cytokines considered responsible for the disorders observed during the course of the disease. PMID- 15899493 TI - Immunohistochemical and ultrastructural findings related to the blood--brain barrier in the blood vessels of the cerebral white matter in aged dogs. AB - Immunohistochemical and ultrastructural analysis of canine brain tissue was performed to determine whether cerebral capillaries, which form the blood--brain barrier (BBB), display age-related morphological changes in the white matter (WM). A slight decrease in laminin immunolabelling was detected in the basement membranes (BMs) of capillaries in the WM of old dogs, as compared with that in the brains of young dogs. The Prussian blue DAB post-DAB enhancement method detected iron present in macrophages and astrocytes in the WM. Copper/zinc superoxide dismutase, MT-I and -II and MT-III immunoreactivity was detected mainly in reactive astrocytes in the WM of aged dogs. Ultrastructurally, collagen like fibrils were detected to a variable degree in the spaces between the BMs of capillary endothelial cells and astrocytes in the WM of some aged dogs. These results suggest that age-related morphological changes in capillaries of the WM are associated with BBB dysfunction, leading to the exudation of serum constituents, including harmful substances (e.g., iron), thereby causing tissue damage by oxidative injury. These factors may play a role in the pathogenesis of severe degenerative changes in the WM of aged dogs. PMID- 15899494 TI - Growth profiles of recent canine distemper isolates on Vero cells expressing canine signalling lymphocyte activation molecule (SLAM). AB - Fresh samples of lymph node, lung and cerebrum taken post mortem from dogs no. 1, 2 and 3 yielded canine distemper virus (CDV) strains 007 Lm, 009 L and 011 C, respectively. These were titrated on Vero cells stably expressing canine signalling lymphocyte activation molecule (SLAM; Vero-DST cells). Growth curves of the three strains were produced by titration of the released virus and cell associated virus at various timepoints. All three isolates, especially 007 Lm, grew well on Vero-DST cells. The titres of cell-associated virus of two strains (009 L and 011 C) were clearly lower than those of virus released into the culture supernate. The results indicate that Vero-DST cells are not only useful for primary isolation but also efficient for titrating virus from fresh tissues and for the study of growth profiles of recent CDV isolates. PMID- 15899495 TI - Immunohistochemical detection of the p27 capsid protein of caprine arthritis encephalitis virus (CAEV) in bone-marrow cells of seropositive goats. AB - Bone-marrow samples were collected from 48 CAEV-seropositive, symptomless goats (30 kids, 18 adults). The samples were formalin-fixed and processed for histological examination. In addition, all samples were examined immunohistochemically with a monoclonal antibody (1A7) against the p27 capsid protein of maedi-visna virus, an antibody which cross-reacts with the Ca-p27 of CAEV. Samples from 16 goats (10/30 kids, 6/18 adults) showed positive immunolabelling of bone-marrow stromal cells (fibrocytes, endothelial cells and adipocytes) and of scattered macrophages, whereas haematopoietic cells were negative. The detection of viral Ca-p27 protein in bone-marrow fibrocytes was consistent with previous in-vitro studies which indicated that such cells are semi-permissive for CAEV infection. It is speculated that bone-marrow stromal cells represent a viral reservoir in symptomless animals. PMID- 15899496 TI - FL-CTL assay: fluorolysometric determination of cell-mediated cytotoxicity using green fluorescent protein and red fluorescent protein expressing target cells. AB - Cytotoxic T lymphocytes (CTLs) are crucial effectors against intracellular pathogens and cancer. Accurate and efficient assessment of CTL activity is important for basic and clinical studies. Widely used CTL assays, including the chromium release, JAM test and ELISPOT, involve either radioisotopes or lengthy procedures. Here, we developed a new fluorolysometric CTL assay based on cell mediated cytolysis of fluorescent protein (GFP or DsRed) expressing cells quantified by one of the fluoro-based methods: flow cytometry, fluorescence microplate reader, or fluorescence microscopy. With flexible detection methods and lentiviral vector transduced stable lines of either GFP+ or DsRed+ cells as targets for antigen presentation and equal number of the other as internal reference for consistency and accuracy, this assay is easy to perform and to scale-up for simultaneous multi-sample analyses. Using two different antigen systems, we demonstrated that this assay is very sensitive to determine primary CTL activity of both in vitro and in vivo primed antigen-specific T cells. Thus, this FL-CTL assay is highly sensitive, reliable, reproducible, economical, convenience and supports broad applications compared to conventional CTL assays. PMID- 15899497 TI - Cuticular hydrocarbon synthesis and its maternal provisioning to embryos in the viviparous cockroach Diploptera punctata. AB - Embryos of the viviparous cockroach Diploptera punctata accumulate large amounts of hydrocarbon (HC) of either maternal or embryonic origin. HC synthesis and its accumulation in maternal and embryonic tissues were measured over the course of gestation. Female abdominal integument was the only tissue that synthesized appreciable amounts of HC in vitro, and did so at an increasing rate from the time of mating to mid-pregnancy, when rates of synthesis declined. The embryos synthesized HC at rates <1% those of the female, showing that the majority of HC detected in and on embryos was of maternal origin. The brood sac that houses the developing embryos did not synthesize HC in vitro, indicating that HC must be transported from the female abdominal integument to the embryos. The mass of female epicuticular HC was constant at approximately 183 microg, while her internal HC increased fourfold from mating to mid-pregnancy, then declined until parturition. The decline in internal HC reflected both declining HC synthesis in the female and greater export to the embryos, as embryonic internal HC increased 250-fold prior to parturition. An external HC coating over the oothecal covering and chorion of the embryos increased to mid-pregnancy, then declined. Unlike oviparous cockroaches, D. punctata females fed throughout the reproductive cycle, reflecting the nutritional demands of continuously provisioning the developing embryos. PMID- 15899498 TI - Neuroprotective strategies in MS: lessons from C57BL/Wld(S) mice. AB - Valuable clues about how axons degenerate in MS can be gained from axon pathology in other disorders and experimental models. We discuss the similarities in mechanism and morphology of axon pathology in diverse circumstances revealed using mutant mice. The slow Wallerian degeneration mutation, Wld(S), delays three types of axon degeneration previously considered distinct: Wallerian degeneration of injured axons, 'dying-back' of axons in peripheral nervous system disease, and axonal spheroid pathology in gracile axonal dystrophy (gad) mice. Therefore, axon degeneration mechanisms are more uniform than previously thought and, in gad at least, axonal swelling is either related to or a consequence of Wallerian degeneration. Both axonal swelling and the accumulation of amyloid precursor protein through impaired axonal transport are common to MS, gad, and many other CNS disorders, indicating a degree of shared mechanism. YFP-H transgenic mice express YFP in a representative subset of neurons enabling unprecedented imaging of axon morphology and pathology over considerable longitudinal distances. Using this method, we have observed unbroken axons with multiple constrictions and dilatations in VEGF(delta/delta) mice, a model of amyotrophic lateral sclerosis (ALS). Similar morphologies have been described in MS, stroke, and other disorders, again suggesting a uniformity of axon degeneration mechanisms. PMID- 15899499 TI - General mechanisms of axonal damage and its prevention. AB - Axonal degeneration is a prominent pathological feature in multiple sclerosis observed over a century ago. The gradual loss of axons is thought to underlie irreversible clinical deficits in this disease. The precise mechanisms of axonopathy are poorly understood, but likely involve excess accumulation of Ca ions. In healthy fibers, ATP-dependent pumps support homeostasis of ionic gradients. When energy supply is limited, either due to inadequate delivery (e.g., ischemia, mitochondrial dysfunction) and/or excessive utilization (e.g., conduction along demyelinated axons), ion gradients break down, unleashing a variety of aberrant cascades, ultimately leading to Ca overload. During Na pump dysfunction, Na can enter axons through non-inactivating Na channels, promoting axonal Na overload and depolarization by allowing K egress. This will gate voltage-sensitive Ca channels and stimulate reverse Na-Ca exchange, leading to further Ca entry. Energy failure will also promote Ca release from intracellular stores. Neurotransmitters such as glutamate can be released by reverse operation of Na-dependent transporters, in turn activating a variety of ionotropic and metabotropic receptors, further exacerbating overload of cellular Ca. Together, this Ca overload will inappropriately stimulate a variety of Ca-dependent enzyme systems (e.g., calpains, phospholipases), leading to structural and functional axonal injury. Pharmacological interruption at key points in these interrelated injury cascades (e.g., at voltage-gated Na channels or AMPA receptors) may confer significant neuroprotection to compromised central axons and supporting glia. Such agents may represent attractive adjuncts to currently available immunomodulatory therapies. PMID- 15899500 TI - Antibiotic resistance and the evolution of group-beneficial traits. II: a metapopulation model. AB - Inspired by the evolution of antibiotic resistance in bacteria, we have developed a model that examines the evolution of "producers" (who secrete a substance that breaks down antibiotics) and non-producers. In a previous study, we found that frequency-dependent selection could favor an intermediate frequency of producers in a single, large population. Here we develop a metapopulation model that examines the evolution of producers and non-producers. Our results indicate that in a metapopulation with many groups, each of size N, the equilibrial frequency of producers decreases with group size. Even when N is high (e.g. 150 individuals/group), however, a significant frequency of producers is still predicted. We also found that the equilibrial frequency of producers increases as the minimum numbers of producers necessary to provide protection to non-producers increases. Lastly, increasing the benefit/cost ratio (b/c) for producers increases their equilibrial frequency. PMID- 15899501 TI - High variation in developmental instability under non-normal developmental error: a Bayesian perspective. AB - The developmental mechanisms behind developmental instability (DI) are only poorly understood. Nevertheless, fluctuating asymmetry (FA) is often used a surrogate for DI. Based on statistical arguments it is often assumed that individual levels of FA are only weakly associated with the underlying DI. Patterns in FA therefore need to be interpreted with caution, and should ideally be transformed into patterns in DI. In order to be able to achieve that, assumptions about the distribution of developmental errors must be made. Current models assume that errors during development are additive and independent such that they yield a normal distribution. The observation that the distribution of FA is often leptokurtic has been interpreted as evidence for between-individual variation in DI. This approach has led to unrealistically high estimates of between-individual variation in DI, and potentially incorrect interpretations of patterns in FA, especially at the individual level. Recently, it has been suggested that the high estimates of variation in DI may be biased upward because either developmental errors are log-normal or gamma distributed and/or low measurement resolution of FA. A proper estimation of the amount (and shape) of heterogeneity in DI is crucial for the interpretation of patterns in FA and their transformation into patterns in DI. Yet, incorrect model assumptions may render misleading inferences. We therefore develop a statistical model to evaluate the sensitivity of results under the normal error model against the two alternative distributions as well as to investigate the importance of low measurement resolution. An analysis of simulated and empirical data sets indicated that bias due to misspecification of the developmental error distribution can be substantial, yet, did not appear to reduce estimates of variation in DI in empirical data sets to a large extent. Effects of low measurement resolution were neglectable. The importance of these results are discussed in the context of the interpretation of patterns in FA. PMID- 15899502 TI - Self-organization of the vascular system in plant leaves: inter-dependent dynamics of auxin flux and carrier proteins. AB - The vegetative hormone Auxin is involved in vascular tissues formation throughout the plant. Trans-membrane carrier proteins transporting auxin from cell to cell and distributed asymmetrically around each cell give to auxin a polarized movement in tissues, creating streams of auxin that presume future vascular bundles. According to the canalization hypothesis, auxin transport ability of cells is thought to increase with auxin flux, resulting in the self-enhancement of this flux along auxin paths. In this study we evaluate a series of models based on canalization hypothesis using carrier proteins, under different assumptions concerning auxin flux formation and carrier protein dynamics. Simulations are run on a hexagonal lattice with uniform auxin production. A single cell located in the margin of the lattice indicates the petiole, and acts as an auxin sink. The main results are: (1) We obtain branching auxin distribution patterns. (2) The type of self-enhancement described by the functional form of the carrier proteins regulation responding to the auxin flux intensity in different parts of a cell, has a strong effect on the possibility of generating the branching patterns. For response functions with acceleration in the increase of carrier protein numbers compared to the auxin flux, branching patterns are likely to be generated. For linear or decelerating response functions, no branching patterns are formed. (3) When branching patterns are formed, auxin distribution greatly differs between the case in which the number of carrier proteins in different parts of a cell are regulated independently, and the case in which different parts of a cell compete for a limited number of carrier proteins. In the former case, the auxin level is lower in veins than in the surrounding tissue, while in the latter, the auxin is present in greater abundance in veins. These results suggest that canalization is a good candidate for describing plant vein pattern formation. PMID- 15899503 TI - Spatial invasion by a mutant pathogen. AB - Imagine a pathogen that is spreading radially as a circular wave through a population of susceptible hosts. In the interior of this circular region, the infection dies out due to a subcritical density of susceptibles. If a mutant pathogen, having some advantage over wild-type pathogens, arises in this region it is likely to die out without leaving a noticeable trace. Mutants that arise closer to the infection wavefront have access to more susceptible hosts and thus are more likely to become established and perhaps (locally) out-compete the original pathogen. Among the factors (position, transmission rate, pathogen induced death rate) that influence the fate of a mutant, which are most important? What does this tell us about the types of mutants that are likely to invade and become established? How do such tendencies serve to steer the evolution of pathogens in a spatial setting? Do different types of models of the same phenomena lead to similar conclusions? We address these issues from the point of view of an individual-based stochastic spatial model of host-pathogen interactions. We consider the probability of a successful invasion by a single mutant as a function of the transmissibility and virulence strengths and the mutant position in the wavefront. Next, for a version of the model in which mutations arise spontaneously, we obtain analytical and simulation results on the mean time to a successful invasion. We also use our model predictions to gain insight into experimental data on bacteriophage plaques. Finally, we compare our results to those based on ordinary and partial differential equations to better understand how different models might influence our predictions on the fate of a mutant pathogen. PMID- 15899504 TI - Agent-based modeling of the context dependency in T cell recognition. AB - Antigen recognition by T cells is a key event in the adaptive immune response. T cells scan the surface of antigen-presenting cells (APCs) or target cells for specific peptides bound to MHC molecules. In the physiological setting, a typical APC presents tens of thousands of diverse endogenous self-derived peptides complexed to MHC (pMHC complexes). When 'foreign' peptides are presented, they constitute a small fraction of the total surface peptide repertoire. As T cells seem to be capable of discerning minute amounts of 'foreign' peptides among a complex background of self-peptides, endogenous peptides are generally assumed to play no role in recognition. However, recent results suggest that these background peptides may alter the sensitivity of T cells to foreign peptides. Current experimental limitations preclude analysis of peptide mixtures approaching physiological complexity, making it difficult to further address the role of complex background peptides. In this paper, we present a computational model to test how complex, varied peptide populations on an APC could potentially modulate a T cell's ability to detect the presence of small numbers of agonist peptides among a diverse population. We use the model to investigate the notion that under physiological conditions, T cell recognition of foreign peptides is context dependent, that is, T cells process signals gathered from all pMHC interactions, not just from a few agonist peptides while ignoring all others. PMID- 15899505 TI - Extracellular superoxide dismutase (EC-SOD) gene mutations screening in a sample of Mediterranean population. AB - The main role of superoxide dismutases (SODs) is to eliminate reactive oxygen species in cells and tissues. Extracellular SOD (EC-SOD/SOD3) is a major superoxide scavenger and it is located on cell surfaces and primarily in extracellular matrix, and binds heparan sulfates by its carboxyterminal portion. Human EC-SOD gene is located on chromosome 4 and comprises three exons and two introns. The SOD3 coding sequence is entirely located within exon 3 and has missense polymorphisms. The Arg213Gly mutation affects the function of the carboxyterminus and correlates with several diseases. In this work, we explored genetic variants within EC-SOD gene of subjects living in southern Italy. Four new variations were detected: one was silent mutation, while three were missense variations that give rise to amino acid substitutions at position 131 (F>C), 160 (V>L) and 202 (R>L) in the mature product. The Arg213Gly variant was not found. The missense mutations in the DNA of assayed 2400 chromosomes had frequencies of 5.34% for the F131C variation, 0.25% for the V160L variation and 0.84% for the R202L variation. The effect of these alterations on the metabolic activity and diseases remains to be further explained. PMID- 15899506 TI - Triphenyltin and Tributyltin inhibit pig testicular 17beta-hydroxysteroid dehydrogenase activity and suppress testicular testosterone biosynthesis. AB - We previously reported that tributyltin chloride (TBT) and triphenyltin chloride (TPT) powerfully suppressed human chorionic gonadotropin- and 8-bromo-cAMP stimulated testosterone production in pig Leydig cells at concentrations that were not cytotoxic [Nakajima Y, Sato Q, Ohno S, Nakajin S. Organotin compounds suppress testosterone production in Leydig cells from neonatal pig testes. J Health Sci 2003;49:514-9]. This study investigated the effects of these organotin compounds on the activity of enzymes involved in testosterone biosynthesis in pig testis. At relatively low concentrations of TPT, 17beta-hydroxysteroid dehydrogenase (17beta-HSD; IC(50)=2.6microM) and cytochrome P450 17alpha hydroxylase/C(17-20) lyase (IC(50)=117microM) activities were inhibited, whereas cholesterol side-chain cleavage cytochrome P450 and 3beta-HSD/Delta(4)-Delta(5) isomerase activities were less sensitive. Overall, TPT was more effective than TBT. TPT also inhibited both ferredoxin reductase and P450 reductase activities at concentrations over 30microM; however, TBT had no effect, even at 100microM. The IC(50) values of TPT were estimated to be 25.7 and 22.8microM for ferredoxin reductase and P450 reductase, respectively. The inhibitory effect of TPT (30microM) on microsomal 17beta-HSD activity from pig testis was eliminated by pretreatment with the reducing agents dithiothreitol (1mM) and dithioerythritol (1mM). On the other hand, TPT (0.03microM) or TBT (0.1microM) exposure suppressed the testosterone production from androstenedione in pig Leydig cells indicating that these organotins inhibit 17beta-HSD activity in vivo as well as in vitro, and the IC(50) values of TPT and TBT for 17beta-HSD activity were estimated to be 48 and 114nM, respectively. Based on these results, it appears possible that the effects of TBT and TPT are largely due to direct inhibition of 17beta-HSD activity in vivo. PMID- 15899507 TI - Osteoprotegerin (OPG) and RANKL expression and distribution in developing human craniomandibular joint. AB - During embryogenesis the bone tissue of craniomandibular joint (CMJ) is formed through two pathways: intramembranous ossification and endochondral ossification. The development process is under the control of regulatory factors. The osteoprotegerin (OPG) and the receptor activator of nuclear factor (NF)-kappaB ligand are key regulators of osteoclastogenesis. The aim of this study is the localization of OPG and RANKL mRNA and protein in the foetal CMJ by immunohistochemistry (IHC) and in situ hybridization (ISH). The main results were: OPG and RANKL mRNA and protein were co-localized in the same cell types; OPG and RANKL were specially immunolocated in osteogenic cells; immunolabeling was often seen in the nucleus and cytoplasm of otherwise negative hypertrophic chondrocytes; IHC and ISH labeling decreased from proliferative to hypertrophic chondrocytes; early osteocytes showed dual protein expression and some of the mature osteocytes were ISH-negative; periosteal osteoclasts and chondroclasts were mostly stained by IHC and variably labeled by ISH; the new bone matrix and trabecular borders showed intense immunolabeling. The co-expression of OPG and RANKL in the same bone cell types confirms their strictly coupled action in the regulation of bone metabolism in the CMJ development and their extracellular presence in the new bone matrix and trabecular borders suggests a local regulatory role. PMID- 15899508 TI - Oogenesis at subzero temperatures: a comparative study of the oocyte morphology in nine species of Notothenioids. AB - Oogenesis was examined in nine species of Antarctic fish to verify the existence of morphological peculiarities. The analyses were carried out on specimens belonging to three different families of Notothenioids (Nototheniidae, Channichthyidae and Bathydraconidae), all captured in the Ross Sea, in front of the Italian Station of Terra Nova Bay. Following dissection, the ovaries were processed and examined at the light and electron microscopes to determine the oocyte gross and fine morphology. The attention, in particular, was focused on the presence of cytoplasmic round bodies and on the organization of the cortical alveoli and the vitelline envelope. Results reveal significant specie-specific differences that could be partly correlated to the phylogenetic radiation but not to the peculiar environmental conditions being essentially comparable to those observed among temperate species. PMID- 15899509 TI - The effects of diethylcarbamazine on the ultrastructure of lung cells in vivo. AB - The pulmonary surfactant synthesis is disturbed in experimentally induced asthma, as are the intracellular storage capacity and its physical activity. These alterations may also be present in chronic asthmatic patients, and therefore the dysfunction of the pulmonary surfactant system may play an important role in the pathophysiology of asthma. Some clinical reports have described favorable results with the use of diethylcarbamazine (DEC) in patients with bronchial asthma showing that DEC is effective in terminating acute attacks of bronchial asthma. The present study aimed to analyze the ultrastructural alterations of lung cells after treatment in vivo with diethylcarbamazine. After 12 days of treatment with DEC, when compared with control samples, type II pneumocytes showed active nuclei with abundant euchromatin and evident nucleoli, and a substantially greater number of mature secretion vesicle. On the other hand, type I pneumocytes showed no morphological alterations. After DEC treatment, lung macrophages also presented several characteristics of cellular activation such as nuclei with a prominence of euchromatin and central nucleoli as well as an abundance of early and late endossomes distributed throughout the cytoplasm. These results confirm that DEC exerts a role in the activation of important pulmonary cellular pathways, which are probably related to the clinical improvement of asthma symptoms after DEC treatment. PMID- 15899510 TI - Effect of counterions on lanthanum biosorption by Sargassum polycystum. AB - The effect of the presence of different anions on the biosorption of La(3+) (Lanthanum) using Sargassum polycystum Ca-loaded biomass was studied in this work. Different types of metal salts were used, such as nitrate, sulphate and chloride. The presence of the anion sulphate decreased the metal uptake for tested pH values of 3--5 when compared to the nitrate and chloride systems. The presence of chloride ions did not seem to interfere with the lanthanum removal. The speciation of lanthanum in solution could explain the differences obtained for the different systems and the Mineql+ program was used for the calculations. A monovalent complex with sulphate and lanthanum was formed that had lower apparent affinity towards the biomass compared to the free trivalent metal ion. The La uptake varied from 0.6 to 1.0 mmol g(-1). The Langmuir model was used to describe quantitatively the sorption isotherms. The addition of sulphuric acid for pH adjustment decreased the metal uptake from lanthanum sulphate solutions when compared to the nitric acid addition. The effect was more pronounced with sulphuric acid due to the formation of complexes. PMID- 15899511 TI - An assessment of anthropogenic source impacts on mercury cycling in the Willamette Basin, Oregon, USA. AB - In Oregon's Willamette River Basin (Basin), methylmercury levels in fish triggered health advisories and required development of a mercury Total Maximum Daily Load (TMDL) for the Willamette River. A seasonally-responsive dynamic systems model is used to identify the principal sources of natural and anthropogenic mercury, the relative contributions of these sources to the river, the impact of hypothetical reductions in specific natural and anthropogenic sources on mercury levels in surface water, sediment, and fish tissue, and the degree to which any such changes would be clearly discernible to environmental managers and Basin stakeholders. Two scenarios are modeled: "PRES", which considered all currently known natural and anthropogenic mercury sources and "LEEM", which (hypothetically) eliminated all local, but not global, anthropogenic sources and greatly lowered native soil erosion rates. Elimination of local air emissions reduces runoff of air-deposited mercury by approximately 34% and advection from the Basin by approximately 12%, while lowering erosion rates reduces particulate runoff by approximately 57%, deposition from the water column to surficial sediment by approximately 33%, and fluvial load by approximately 24%; for a net reduction of 25.6% in the total mercury load to the river. Such hypothetical reductions bring methylmercury concentrations in predatory fish to levels that would allow restoration of fish consumption as a beneficial use. However, several factors, primarily technical feasibility and global sources, may impede attempts to attain this beneficial use. Actualizing the hypothetical 100% elimination of local anthropogenic sources and a >50% reduction in erosion could pose significant technical challenges. Because local anthropogenic emissions make relatively smaller contributions to the Basin than do persistent global sources (sources over which there is little, if any, possibility of local control), localized environmental management actions alone may not be adequate to address mercury impacts within the Basin. PMID- 15899512 TI - Provider preferences for DVT prophylaxis. AB - BACKGROUND: Patients admitted to medicine wards are typically elderly with multiple co-morbidities, placing them at significant risk for venous thromboembolic (VTE) disease. Standard guidelines to help physicians assess risk and institute prophylaxis do not currently exist for this patient population. Our purpose was to assess how clinicians would rate a patient's risk for VTE and what recommendations they would make for prophylaxis. METHODS: Internal medicine residents and staff at a tertiary care medical center were asked to identify risk factors, evaluate VTE risk, and recommend a method of prophylaxis for patients described in eight clinical vignettes created by the authors. Each vignette was designed to represent a patient at a specific level of risk. RESULTS: 35 physicians returned the survey. Responders were able to identify some risk factors and weigh them appropriately when assigning an overall risk level. There was good agreement on risk level among responders (k=0.62) and moderate agreement comparing responders with our pre-defined gold standard (GS) (k=0.42). Compared to the GS they underestimated the level of risk almost 50% of the time. The risk level they assigned affected the type of prophylaxis recommended, with fewer low risk patients receiving any type of heparin and more high risk patients receiving low-molecular weight heparin (LMWH). CONCLUSIONS: Although internal medicine physicians are able to identify some risk factors for in-hospital VTE, they consistently underestimate the overall risk, leading to less aggressive preventative measures. Continuing education is essential to combating this preventable inpatient complication. PMID- 15899513 TI - Adaptogenic and anti-amnesic properties of Evolvulus alsinoides in rodents. AB - Evolvulus alsinoides (EA) is well known for its memory enhancement, antiepileptic and immunomodulatory properties in the traditional Indian system of medicine, Ayurveda. In view of the increasing attention towards plants offering non specific resistance (adaptogens) towards stress, we have evaluated crude ethanolic extract of EA for its adaptogenic and memory enhancing properties in rodents. Adaptogenic activity was assessed in rats subjected to acute and chronic unpredictable stress. Male Sprague-Dawley rats, weighing 180-200 g were immobilized for 150 min once only in acute stress (AS) model, whereas in chronic unpredictable stress (CUS) model rats were subjected to different types of stressors daily for 7 days. Stress exposure has induced gastric ulceration with increase in adrenal gland weight, plasma creatine kinase (CK), and corticosterone level in AS and CUS. However plasma glucose was increased only in AS. Rats were treated with graded doses of crude ethanolic extract of EA (100, 200 and 400 mg/kg p.o.) for 3 days and subjected to AS on 3 day after 45 min of last dose. In CUS, EA at a dose of 200 mg/kg p.o. found effective in acute studies was administered 45 min prior to stress regimen for 7 days. EA reduced the stress induced perturbations similar to Panax quinquefolium (PQ) (100 mg/kg p.o.), a well known adaptogen. EA (100 mg/kg) administered orally for 3 days in adult male Swiss mice, was effective in decreasing scopolamine induced deficit in passive avoidance test. The improvement in the peripheral stress markers and scopolamine induced dementia by EA in the present study indicates the adaptogenic and anti amnesic properties of EA. PMID- 15899514 TI - Monocyte viability on titanium and copper coated titanium. AB - The role of apoptosis/cell death in the inflammatory response at the implanted materials is unexplored. Two surfaces with different cytotoxic potential and in vivo outcomes, titanium (Ti) and copper (Cu) were incubated in vitro with human monocytes and studied using a method to discriminate apoptotic and necrotic cells (Annexin V/PI staining). Further, staurosporine, a potent inducer of apoptosis, was added to the surface adherent monocytes. Lactate dehydrogenase (a marker of cell membrane injury) and TNF-alpha and IL-10, cytokines, previously suggested to play a major role in the monocyte apoptosis, were assayed in the culture medium. The results demonstrated that Ti surfaces displayed enhanced monocyte survival and production of IL-10 and TNF-alpha. Cu adherent cells exhibited apoptotic signs as early as 1h after incubation. In contrast to Ti, after 48 h the predominance of apoptotic cells switched to apoptotic/necrotic cells on Cu surfaces. Staurosporine treatment of Ti adherent cells mediated similar type of cell death. LDH and cytokine contents were low around Cu surfaces, partly explained by interference between Cu ions and LDH and cytokines. This study suggests that material properties rapidly influence the onset of human monocyte apoptosis and progression to late apoptosis/necrosis. Early detection of apoptosis and cell death may be important for the understanding of the biological response to implanted materials. PMID- 15899515 TI - The effect of RGD peptide-conjugated magnetite cationic liposomes on cell growth and cell sheet harvesting. AB - Tissue engineering requires novel technologies for establishing 3D constructs, and the layered method of culturing cell sheets (cell sheet engineering) is one potentially useful approach. In the present study, we investigated whether coating the culture surface with RGD (Arg-Gly-Asp) peptide-conjugated magnetite cationic liposomes (RGD-MCLs) was able to facilitate cell growth, cell sheet construction and cell sheet harvest using magnetic force without enzymatic treatment. To promote cell attachment, an RGD-motif-containing peptide was coupled to the phospholipid of our original magnetite cationic liposomes (MCLs). The RGD-MCLs were added to a commercially available 24-well ultra-low-attachment plate the surface of which comprised a covalently bound hydrogel layer that was hydrophilic and neutrally charged. A magnet was placed on the underside of the well in order to attract the RGD-MCLs to the surface of the well, and then NIH/3T3 cells were seeded into the well. Cells adhered to the bottom of the culture surface, which was coated with RGD-MCLs, and the cells spread and proliferated to confluency. After incubation, the magnet was removed and the cells were detached from the bottom of the plates, forming a contiguous cell sheet. Because the sheets contained magnetite nanoparticles, they could be harvested using a magnet inserted into the well. These results suggest that this novel methodology using RGD-MCLs and magnetic force, which we have termed 'magnetic force-based tissue engineering (Mag-TE)', is a promising approach for tissue engineering. PMID- 15899516 TI - Impaired breast cancer resistance protein mediated drug transport in plasma cells in multiple myeloma. AB - The breast cancer resistance protein (BCRP/ABCG2) is an ATP-binding-cassette transporter involved in the transport of drugs used in the treatment of multiple myeloma (MM). Its expression, function and clinical significance in MM, however, are unknown. We report that BCRP is preferentially expressed and functionally active in normal plasma cells but that its function is significantly impaired in plasma cells in newly diagnosed MM. The data presented argue against a role for BCRP in primary drug resistance in MM and the utilisation as a molecular target as such but warrant research into its (patho)physiological role in normal and malignant plasma cells. PMID- 15899517 TI - Listening to mutant mice: a spotlight on the role of CRF/CRF receptor systems in affective disorders. AB - Genetically engineered mice were originally generated to delineate the role of a specific gene product in behavioral or neuroendocrine phenotypes, rather than to produce classic animal models of depression. To learn more about the neurobiological mechanisms underlying a clinical condition such as depression, it has proven worthwhile to investigate changes in behaviors characteristic of depressed humans, such as anxiety, regardless of whether or not these alterations may also occur in other disorders besides depression. The majority of patients with mood and anxiety disorders have measurable shifts in their stress hormone regulation as reflected by elevated secretion of central and peripheral stress hormones or by altered hormonal responses to neuroendocrine challenge tests. In recent years, these alterations have been increasingly translated into testable hypotheses addressing the pathogenesis of illness. Refined molecular technologies and the creation of genetically engineered mice have allowed to specifically target individual genes involved in regulation of corticotropin releasing factor (CRF) system elements (e.g. CRF and CRF-related peptides, their receptors, binding protein). Studies performed in such mice have complemented and extended our knowledge. The cumulative evidence makes a strong case implicating dysfunction of these systems in the pathogenesis of depression and leads us beyond the monoaminergic synapse in search of eagerly anticipated strategies to discover and develop better therapies for depression. PMID- 15899518 TI - Mouse splenic B lymphocyte activation using different activation stimuli induces in vitro splicing of tumor necrosis factor-alpha nuclear pre-mRNA. AB - The pleiotropic functions of tumor necrosis factor-alpha (TNFalpha) have brought considerable attention in the past decade to its physiological and pathological roles in inflammatory and autoimmune diseases. However, little is known about how the production of TNFalpha is regulated at the transcriptional and translational levels in immune cells such as T and B lymphocytes. Our previous study demonstrated that unspliced "pre-mRNA" of TNFalpha is present in resting T cells. Initiation of splicing of TNFalpha pre-mRNA to mature mRNA requires T cell activation, which is unique and necessary for TNFalpha production when compared to its production in mononuclear phagocytes, including different lineages of macrophages (Mvarphi) and dendritic cells (DC). In this study, we further demonstrate that resting mouse B cells also contain pre-existing TNFalpha mRNA. The physiological process of B cell activation induced by (1) either the cross linking of the B cell receptor (BCR) or CD40, (2) treatment with LPS, or PMA plus ionomycin, induces TNFalpha mRNA splicing in vitro. The kinetic response of TNFalpha splicing in B cells is much slower when compared to that in activated T cells. Studies using well-known kinase inhibitors demonstrated that MAP kinase kinase (MEK) and protein kinase C (PKC) are required for TNFalpha splicing upon stimulation through the BCR. These studies demonstrate that the production of TNFalpha in activated B cells is regulated differently than in activated T cells, and these differences may allow for the selective inhibition of TNFalpha in various autoimmune diseases depending on the mechanism of action of the selected anti-TNFalpha therapy. PMID- 15899519 TI - Molecular characterization of L-type calcium channel splice variants expressed in human T lymphocytes. AB - Calcium (Ca2+) influx is a fundamental intracellular signal that is required to initiate and sustain T lymphocyte activation. Dihydropyridine-sensitive, L-type Ca2+ channels appear to play a significant role in Ca2+ mobilization during T cell activation, but very little is known about the molecular structure of these channels in T lymphocytes. Here we identify two novel splice variants of the Ca(V)1.4 (alpha1F) L-type Ca2+ channel that are expressed in human T lymphocytes, and also demonstrate expression of the Ca(V)1.4 protein in the human Jurkat T cell leukemia line and human peripheral blood T lymphocytes (PBTs). The carboxy termini of both Ca(V)1.4 splice isoforms contain unique exon usages distinct from the Ca(V)1.4 channel isolated from human retina that may render these channel variants insensitive to changes in membrane depolarization. Additional evidence of the importance of these new splice variants comes from the demonstration that the mRNA expression of the Ca(V)1.4 splice isoforms is regulated by TCR-induced activation in Jurkat T cells, and to a lesser extent in human PBTs. Overall these results provide the first evidence that structurally unique L-type Ca2+ channels exist in T lymphocytes, which can contribute to a Ca2+ influx during T lymphocyte activation. PMID- 15899520 TI - 17beta-estradiol protects SH-SY5Y Cells against HIV-1 gp120-induced cell death: evidence for a role of estrogen receptors. AB - Despite the large body of experimental evidence demonstrating the neuroprotective properties of 17beta-estradiol (17beta-E2) both in vitro and in vivo experimental models of neuronal injury, the exact mechanisms implicated in neuroprotection have not been fully delineated. Some experimental evidence highlight a role for the antioxidant properties of 17beta-E2 in mediating protection against oxidative injury. Parallel to these, evidence also exist which point to alternative mechanisms involving estrogen receptors (ER). The HIV-1 coat protein, gp120, has been implicated in the progression of central nervous system damage caused by HIV 1 infection. The neurotoxic effects induced by gp120 are triggered via an excitotoxic mechanism of cell death which implicates alteration of calcium homeostasis, activation of calcium-dependent pathways, mitochondrial uncoupling and membrane lipid peroxidation. In the present study, we demonstrate that 17beta E2 protects human SH-SY5Y neuroblastoma cells from cell death elicited by gp120. Tamoxifen and ICI 182,780, two ER antagonists, both antagonized 17beta-E2 mediated inhibition of cell death. Exposure of SH-SY5Y cells to gp120 for 30min caused a significant accumulation of intracellular reactive oxygen species (ROS) and this was abrogated by 17beta-E2; however, the ability of 17beta-E2 to counteract ROS generation induced by gp120 does not account for the reported prevention of cell death because ICI 182,780 failed to revert intracellular ROS reduction caused by 17beta-E2 though it was able to revert prevention of cell death. Furthermore, by using 17alpha-E2, the isomer unable to stimulate ER which, however, retains the antioxidant effects, we observed that a pre-treatment with 17alpha-E2 was effective in preventing gp120-induced accumulation of ROS but it failed to affect cell death caused by the viral protein. Collectively, these data demonstrate that neuroprotection afforded by 17beta-E2 is receptor-mediated and ROS scavenging effects may not be implicated. PMID- 15899521 TI - Homology modelling of the major peanut allergen Ara h 2 and surface mapping of IgE-binding epitopes. AB - Three-dimensional models built for the peanut Ara h 2 allergen and other structurally-related 2S albumin allergens of dietary nuts exhibited an overall three-dimensional fold stabilized by disulphide bridges well conserved among all the members of the 2S albumin superfamily. Conformational analysis of the linear IgE-binding epitopes mapped on the molecular surface of Ara h 2 showed no structural homology with the corresponding regions of the walnut Jug r 1, the pecan nut Car i 1 or the Brazil nut Ber e 1 allergens. The absence of epitopic community does not support the allergenic cross-reactivity observed between peanut and walnut or Brazil nut, which presumably depends on other ubiquitous seed storage protein allergens, namely the vicilins. However, the major IgE binding epitope identified on the molecular surface of the walnut Jug r 1 allergen shared a pronounced structural homology with the corresponding region of the pecan nut Car i 1 allergen. With the exception of peanut, 2S albumins could thus account for the IgE-binding cross-reactivity observed between some other dietary nuts, e.g. walnut and pecan nut. PMID- 15899522 TI - Alterations of self-reactive antibody repertoires in HIV disease: an insight into the role of T cells in the selection of autoreactive B cells. AB - Infection with human immunodeficiency virus (HIV) is characterized by a progressive depletion of CD4(+) T cells that parallels a dysfunction of the B cell compartment and a disturbed recognition of self-antigens. The relationship between T lymphocyte homeostasis and abnormalities in the selection of self reactive B cells is not clear as yet. We have therefore compared repertoires of natural antibodies of healthy donors and of patients at various stages of HIV infection. The reactivity of IgM and IgG antibodies in plasma of healthy blood donors and of HIV-positive patients with high and low CD4(+) T cell counts was assessed by semi-quantitative immunoblotting using self-antigens extracted from normal human tissues. Repertoires of reactivites were compared between groups of individuals by means of multiparametric statistical analysis. We observed that repertoires of self-reactive IgM and IgG from HIV-seropositive patients exhibited significantly altered patterns of reactivity, as compared to those of healthy controls. Further, self-reactive repertoires of IgM and IgG of patients with high CD4(+) T cell counts differed significantly from those of patients with low CD4(+) T cell counts. A longitudinal analysis of self-reactive antibody repertoires of progressor and non-progressor patients suggested an influence of CD4(+) T cell counts on immunoglobulin reactivity toward self-antigens. These observations support the hypothesis that altered T cell/B cell interactions due to altered CD4(+) T cell help severely impact on the selection of self-reactive antibody repertoires and may contribute to the onset of pathological autoimmunity in HIV disease. PMID- 15899523 TI - Mobilisation of specific T cells from lymph nodes in contact sensitivity requires substance P. AB - Capsaicin-mediated depletion of neuropeptides in the skin was previously shown to abolish a dinitrocholorobenzene (DNCB)-induced contact sensitivity (CS) response. To understand the basis for this disruption, we explored whether nerve fibres innervating the draining lymph node (LN) could be involved. As expected, removal of the draining LN after DNCB sensitisation abolished the CS response. Furthermore, the CS response could be abolished by destroying the nerve fibres in the draining LN and could be restored by providing the LN with the neuropeptide substance P. The size of the CS response restored by substance P was dose dependent. The response was also inhibited by exposing the lymph node to a neurokinin-1 receptor antagonist which blocks binding of substance P. The results suggest that an afferent signal from the skin via the sympathetic arm of the central nervous system evokes an efferent signal to the LN which combines to regulate the CS response. The efferent signal may serve to control or release from the LN primed effector lymphocytes into the circulation. PMID- 15899524 TI - Epicutaneously induced TGF-beta-dependent tolerance inhibits experimental autoimmune encephalomyelitis. AB - Multiple sclerosis (MS) is a devastating autoimmune disorder of the central nervous system (CNS) with limited treatment modalities. To evaluate the efficacy of epicutaneous (e.c.) tolerance induction in the prevention of CNS autoimmunity, we utilized an animal model of multiple sclerosis: experimental autoimmune encephalomyelitis (EAE). We show that application of myelin basic protein (MBP) to the skin prior to the induction of EAE by immunization with MBP protected mice from developing disease. In addition, e.c. application of MBP at the first clinical signs of EAE, also resulted in suppression of disease. This therapeutic effect was transferable to naive recipients with lymph node cells from MBP treated mice. These regulatory cells were found to be antigen non-specific, as suppression of EAE also occurred when the foreign antigens OVA or TNP were e.c. applied. The mechanistic basis for the tolerance was found to be the production of TGF-beta by the antigen induced toleragenic regulatory T cells. These data demonstrate that e.c.-induced regulatory T cells are potent inhibitors of antigen specific T cell responses, and suggest that e.c. tolerization may have potential effectiveness in the treatment of autoimmune disorders. PMID- 15899525 TI - A rapid and accurate assay for assessing the cytotoxicity of viral proteins. AB - A fluorescence-based assay is presented for measuring the cytoxicity of viral proteins added exogenously to cells. The assay is based on the use of two fluorescent dyes, calcein-AM and ethidium homodimer (EtD-1) to specifically stain living and dead cells respectively and employs fluorescence activated cells sorting (FACS) to achieve a rapid and accurate measurement of the cytotoxic capacity of a potential viral toxin. The assay has been developed using the group B homologue (ADRV-NSP4) of the NSP4 enterotoxin encoded by Group A rotaviruses but should be applicable to assaying any viral protein exhibiting cytotoxic activity. PMID- 15899526 TI - Identification of a multixenobiotic resistance mechanism in primary cultured epidermal cells from Oncorhynchus mykiss and the effects of environmental complex mixtures on its activity. AB - Multixenobiotic resistance (MXR) is a mechanism analogous to the mammalian multidrug resistance (MDR) phenotype, whereby, simultaneous resistance is conferred against the intracellular accumulation of structurally and functionally diverse, natural, endogenous and environmental toxicants. Expression of P glycoproteins (P-gp), ATP-dependent transporters encoded for by the mdr1 gene that have been implicated in this xenobiotic efflux mechanism, have previously been detected in normal teleost tissues involved in a secretory, absorption or a barrier function. The presence of these proteins in the epidermis of fish species has not to our knowledge previously been investigated. In the present study, primary cultures of epidermis from the rainbow trout Oncorhynchus mykiss were employed to investigate whether an MXR mechanism is functional in the epidermis of fish. The efflux of the fluorescent mdr1 substrate rhodamine 123 from the cells was significantly inhibited by verapamil, a compound known to interfere with P-gp mediated transport. The cultured epidermal cells were also observed to accumulate this fluorescent dye in a verapamil sensitive manner, thus indicating the presence of an mdr1-like mechanism. Immunocytochemical analysis, using a monoclonal antibody (JSB1) directed against a conserved cytoplasmic P-gp epitope, also demonstrated the presence of P-gp-like proteins. Sediment elutriate extracts were employed as models of environmental complex mixtures to evaluate the potential of the epidermal cultures to discriminate between samples of varying contaminant burden using MXR activity as an endpoint. The induction of P-gp expression was found to be in accordance with the level of contamination detected in the sediments from which the elutriates were extracted. The findings of the functional study also demonstrated that environmental pollutants, which interfere with P-gp function, could be identified using this model. PMID- 15899527 TI - Toxicity testing of herbicide norflurazon on an aquatic bioindicator species--the planarian Polycelis felina (Daly.). AB - Norflurazon is a bleaching, preemergence herbicide. Due to its mobility and long half-life it presents a potential for groundwater contamination. The aim of our study was to investigate toxic effects of norflurazon on non-targeted aquatic bioindicator organism, the planarian Polycelis felina (Daly.). Animals were exposed to water solutions of norflurazon in concentrations 200, 20, 2 and 0.2 microM. Mortality, locomotive and morphological changes were monitored. Histological changes were studied both on treated and control animals with light microscopy. The primary DNA damage on single planarian cells was studied using the alkaline comet assay. Three comet parameters were studied: tail length, percentage of DNA in comet tail and tail moment. The results showed that norflurazon caused mortality, locomotive, morphological and histological changes in treated animals compared to corresponding controls. The most prominent histological changes were damage of the outer mucous layer, lack of rhabdites, damage to epidermis and extensive damage to parenchyma cells. The results of alkaline comet assay indicated that norflurazon in concentrations of 2 and 0.2 microM induces significant increase of primary DNA damage in planarian cells compared to the corresponding control animals. The mean values of all three measured parameters were significantly elevated on the fourth day of the treatment compared with the first and the seventh day. Based on the results of mortality and locomotive observations, we conclude that the fourth day of the treatment represents a certain threshold within planarian metabolism followed by the beginning of detoxification and recovery. However, histological preparations and comet data statistics show results indicating that high toxicity on the seventh day of the treatment gave the results of decrease of DNA damage due to the tissue/cell damage (apoptosis) and not recovery. The present study showed the ability of norflurazon to induce a wide range of different toxicological responses in freshwater planarian Polycelis felina (Daly.). PMID- 15899528 TI - Biochemical characterization of the two nucleosome assembly proteins from Plasmodium falciparum. AB - The human malaria parasite Plasmodium falciparum contains two nucleosome assembly proteins, which we have termed PfNAPS and PfNAPL. We have over-expressed, purified and characterized these proteins using biochemical and biophysical techniques. PfNAPS and PfNAPL exist as dimers in solution and circular dichroism studies suggest that they may have different three-dimensional protein structures. ELISA-based binding data also suggest that PfNAPS and PfNAPL preferentially interact with the H3-H4 tetramer histones over H2A and H2B histones. We show that the parasite lysate phosphorylates only PfNAPL and this phosphorylation can be inhibited by heparin suggesting a potential role of casein kinase II in this process. Immuno-fluorescence experiments revealed that both PfNAPS and PfNAPL were expressed in all erythrocytic stages of the parasite. PfNAPL was predominantly localised in the cytoplasm in asexual and sexual stages of the parasite. PfNAPS did not co-localise with PfNAPL and was more intimately associated with the parasite nucleus, most strikingly in P. falciparum gametocytes. Taken together, our data show that although PfNAPS and PfNAPL share histone chaperone acitivities, they are regulated differently by phosphorylation and are spatially segregated within the parasite. These proteins are therefore likely to play non-redundant roles as nucleosome assembly motors in the parasite. PMID- 15899529 TI - The problem with advance directives: maybe it is the medium, not the message. AB - Some of today's most significant bioethical challenges center around decisions to initiate or withhold medical treatment for incapacitated patients. In order to ascertain what treatment the patient would have desired, physicians often rely on written advance directives and designated surrogate decision-makers. Unfortunately, both approaches suffer from numerous shortcomings that ultimately limit their usefulness. Although several strategies have been proposed to improve their value, problems nevertheless remain when relying upon written advance directives. We submit that the problem is the medium, not the message-that written advance directives and/or reliance on surrogate decision-makers are fundamentally inadequate. We hypothesize that videotaped advance directives (VADs) can better communicate the specifics, depth, strength and passion of a patient's wishes, more closely approximating the communication that occurs when a physician discusses these issues directly with a patient. VADs may thus enhance the physician's understanding of the patient's wishes. VADs may also ease family conflict and save physician's considerable time by helping family members reach a stronger consensus on the patient's wishes, and do so in a timelier manner. This article reviews the limitations of written advance directives and surrogate decision-makers and describes why VADs may be helpful in overcoming these limitations. PMID- 15899530 TI - Cultivation of high-rate sulfate reducing sludge by pH-based electron donor dosage. AB - A novel self-regulating bioreactor concept for sulfate reduction is proposed aiming for high biomass concentrations and treatment capacities. The system consists of a cell suspension of sulfate reducing bacteria in a continuous stirred tank reactor (30 degrees C) fed with a mixture of both electron donor and electron acceptor (formic acid and sulfuric acid, respectively), nutrients and phosphate buffer via a pH controller. The pH rise due to sulfate reduction is balanced with dosage of the sulfate reducing substrates as acids. The reactor concept was shown to be capable of full sulfate reduction without competition for the electron donor by methanogens and acetogens. Activity assays revealed that hardly any methanogenic activity on formate was left in the suspension by the end of the continuous run (130 days). In addition, the sulfidogenic activity with formate and H2/CO2 had increased, respectively, 3.9 and 11.6 times at the end of the experimental run. The evolution of the particle size distribution of the cell suspension over time indicated that newly grown cells have the tendency to attach together in flocs or to the existing agglomerates. PMID- 15899531 TI - Lipase-catalyzed biosynthesis of cinnamoylated lipids in a selected organic solvent medium. AB - Biosynthesis of cinnamoylated lipids through the lipase-catalyzed transesterification reaction of cinnamic acid with triolein was investigated in organic solvent media. Electrospray ionization-mass spectroscopy (ESI-MS) structural analysis of the reaction mixture revealed the formation of two major end products, monoleyl-1(3)-cinnamate and dioleyl-2-cinnamate. Decreasing the molar ratio of cinnamic acid to triolein from 1:1 to 1:4.5 resulted in an increase in the maximum bioconversion yield of cinnamoylated lipids from 19 to 42%, which remained constant at a lower ratio of 1:6. However, an excess of triolein appeared to have a more beneficial effect on the formation of dioleyl-2 cinnamate than monoleyl-1(3)-cinnamate, leading to different end product compositions at ratios of substrates. With cinnamic acid to triolein ratios of 1:4.5 and 1:6.0, an increase in the bioconversion yield of cinnamoylated lipids to 55% was achieved by adding 2.2 mgmL(-1) silica gel to the reaction mixture. Radical scavenging activity of cinnamoylated lipids, with 50% of radical 2,2 diphenyl-1-picrylhydrazyl (DPPH) scavenging, was found to be higher than that of its corresponding phenolic acid. PMID- 15899532 TI - Microbial adaptation to biodegradation of tert-butyl alcohol in a sequencing batch reactor. AB - This study demonstrates the utility of the sequencing batch reactor (SBR) to adapt microorganisms towards biological removal of tert-butyl alcohol (TBA). The reactor was inoculated with activated sludge and fed with TBA as the sole carbon source. Start-of-cycle TBA concentrations were initially set at 100 mgL(-1) with a cycle time of 24 h and a volumetric exchange ratio of 50% to maintain a TBA loading rate of not more than 100 mgL(-1)d(-1). Step increases in TBA loading rates up to 600 mgL(-1)d(-1) were achieved by first raising the start-of-cycle TBA concentration to 150 mgL(-1) on day 90 and subsequently by reducing the cycle time from 24 to 12, 8 and 6h on days 100, 121 and 199, respectively. This acclimation strategy favored the retention of increasingly higher densities of well-adapted microbial populations in the reactor. The increases in TBA loading produced better settling biomass and higher biomass concentrations with higher specific TBA biodegradation rates. Effluent TBA concentrations were consistently below the detection limit of 25 microgL(-1). The use of progressively shorter cycle times created selection pressures that fostered the self-immobilization of the reactor microorganisms into aerobic granules which first appeared on day 125. Specific TBA biodegradation rates in the granules followed the Haldane model for substrate inhibition, and peaked at 13.8 mgTBAgVSS(-1)h(-1) at a TBA concentration of 300 mgL(-1). Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes from granules sampled between days 220 and 247 confirmed the existence of a highly stable microbial community with members belonging to the alpha, beta and delta subdivisions of Proteobacteria and the Cytophaga-Flavobacteria-Bacteroides (CFB) group. PMID- 15899533 TI - Volatile and polar compounds in Rosadamascena Mill 1803 cell suspension. AB - Studies were conducted on low molecular metabolites (volatiles and polar compounds) produced by Rosa damascena Mill 1803 cell suspension culture, cultivated under different regimes: as a free suspension (in flasks and in bioreactor) and in a two-phase system (in the presence of Amberlite XAD-4 as a second phase). It was established that the main groups of volatiles were hydrocarbons and free acids and their esters and only traces of terpenoids were found. The main components of polar fraction were free acids, especially amino acids and oxidized acids. Depending on the culture conditions, significant differences were established in the amounts of all compounds under study in biomasses, culture media and adsorbed on the second phase (Amberlite XAD-4). PMID- 15899534 TI - Impact of the interaction between adenovirus E1A and CtBP on host cell gene expression. AB - In cell lines harbouring inducible adenovirus E1A genes, the cytotoxicity of wild type E1A was manifested by poor and subsiding expression of the E1A protein during prolonged induction. In contrast, cells expressing E1A deleted in the C terminal binding protein (CtBP)-interaction domain (E1ADeltaCID) demonstrated high levels of expression for extended time. Microarray analyses of host cell gene expression demonstrated that approximately 70% of the regulated genes were increased upon E1A induction and that the majority of E1A-regulated genes were similarly regulated by wild type E1A and E1ADeltaCID. However, for 29 genes, regulation by wild type E1A and E1ADeltaCID were different. Consistent with the altered transforming capacity of E1A unable to bind CtBP, genes involved in tumour cell progression and growth suppression were found among the differently regulated genes. Moreover, promoter sequences of genes up regulated by wild type E1A and/or repressed by E1ADeltaCID demonstrated a higher prevalence of potential binding sites for the CtBP-targeted transcription factors Ets, Ikaros and/or partial differentialEF1/ZEB, suggesting that the failure to block CtBP-repression contributed to the "hyper-transforming" phenotype of E1ADeltaCID. Since E1ADeltaCID also specifically activated host cell gene expression, we find it likely that additional, possibly CtBP-independent, mechanisms contribute to the altered phenotype of E1ADeltaCID-expressing cells. PMID- 15899535 TI - Principles and indicators of successful consumer involvement in NHS research: results of a Delphi study and subgroup analysis. AB - Consumer involvement in NHS research is Department of Health policy within the UK. Despite the existence of policy directives and guidance, until recently there has been no consensus among consumers and researchers about what it means to involve consumers successfully in NHS research. This paper discusses the value of consensus research in this policy area, and presents the detailed findings of a Delphi study carried out to reach consensus on principles and indicators of successful consumer involvement in NHS research. Study participants, comprising consumers, researchers and consumer-researchers, were identified using a purposive sampling strategy. Consensus was reached on eight clear and valid principles of successful consumer involvement in NHS research, with each principle having at least one clear and valid indicator. Subgroup analysis revealed few significant differences in how consumers, researchers and consumer researchers rated the principles and indicators. The implications and limitations of the study are discussed. Further research is needed to assess: (1) the usefulness of the principles and indicators for differing models of consumer involvement, health research methodologies, and subject areas within health research; and (2) the impact of 'successful' consumer involvement on health research processes and outcomes. PMID- 15899536 TI - Paper tigers--do clinical guidelines improve health care quality in patients with testicular germ cell tumors in Germany? AB - In Germany, germ cell tumors are characterized by a marked excess mortality, i.e. a discrepancy between the actual mortality and the mortality expected with adequate implementation of the standard therapy. Moreover, Germany not only has a significantly increased mortality in an international comparison but also shows marked regional differences in the quality of medical care. This is partly due to difficulties in implementing the standard therapy. An attempt was made to improve the quality of medical care by setting up evidence-based diagnostic and therapeutic guidelines in 1996. Contrary to expectations, however, these guidelines have thus far remained largely ineffective, their implementation being hampered by a solely passive dissemination strategy. Added to this are inadequate medical care structures in which the guidelines cannot be properly implemented because of the organizational, social and professional context. Decisive for a quality improvement in the diagnosis, therapy and care of germ cell tumor patients is active dissemination of the guidelines supplemented by a change in the care structure. It is also important to establish indicators for measuring the success of guideline implementation and to perform continuous progress monitoring in order to specifically overcome evident barriers. Future research is required to create a better theoretical basis and to develop further strategies for guideline dissemination and implementation. PMID- 15899537 TI - Near-infrared spectroscopy and imaging: basic principles and pharmaceutical applications. AB - Near-infrared (NIR) spectroscopy and imaging are fast and nondestructive analytical techniques that provide chemical and physical information of virtually any matrix. In combination with multivariate data analysis these two methods open many interesting perspectives for both qualitative and quantitative analysis. This review focuses on recent pharmaceutical NIR applications and covers (1) basic principles of NIR techniques including chemometric data processing, (2) regulatory issues, (3) raw material identification and qualification, (4) direct analysis of intact solid dosage forms, and (5) process monitoring and process control. PMID- 15899538 TI - Negative impact of hypocaloric feeding and energy balance on clinical outcome in ICU patients. AB - BACKGROUND AND AIMS: Critically ill patients with complicated evolution are frequently hypermetabolic, catabolic, and at risk of underfeeding. The study aimed at assessing the relationship between energy balance and outcome in critically ill patients. METHODS: Prospective observational study conducted in consecutive patients staying > or = 5 days in the surgical ICU of a University hospital. Demographic data, time to feeding, route, energy delivery, and outcome were recorded. Energy balance was calculated as energy delivery minus target. Data in means+/-SD, linear regressions between energy balance and outcome variables. RESULTS: Forty eight patients aged 57+/-16 years were investigated; complete data are available in 669 days. Mechanical ventilation lasted 11+/-8 days, ICU stay 15+/-9 was days, and 30-days mortality was 38%. Time to feeding was 3.1+/-2.2 days. Enteral nutrition was the most frequent route with 433 days. Mean daily energy delivery was 1090+/-930 kcal. Combining enteral and parenteral nutrition achieved highest energy delivery. Cumulated energy balance was between 12,600+/-10,520 kcal, and correlated with complications (P < 0.001), already after 1 week. CONCLUSION: Negative energy balances were correlated with increasing number of complications, particularly infections. Energy debt appears as a promising tool for nutritional follow-up, which should be further tested. Delaying initiation of nutritional support exposes the patients to energy deficits that cannot be compensated later on. PMID- 15899539 TI - Vaccination technique, PPD reaction and BCG scarring in a cohort of children born in Guinea-Bissau 2000-2002. AB - The rates of positive tuberculin skin test (TST) reactions and BCG scarring after BCG vaccination vary between studies and populations. Tuberculin reactivity and BCG scarring may be related to better child survival in low-income countries. We therefore studied determinants for TST reaction and scarring in Guinea-Bissau. In a cohort of children born in suburban Bissau from March 2000 to July 2002, we assessed a Mantoux test with Purified protein derivative (PPD) (SSI, 2 T.U.) at 2 (2689 children), 6 (N=2148) and 12 months (N=1638) of age, and BCG scar was assessed at 2 (N=2698) and 6 months (N=2225) of age. In a subgroup of the children the vaccination technique was monitored by direct observation of post vaccination wheal and route of administration. Three different types of BCG vaccine supplied by the local Extended Programme on Immunization were used. At 6 months of age the rate of PPD reactors (>1mm) after BCG vaccination was 25% and the rate of scarring was 89%. One BCG strain was associated with fewer PPD reactors (OR=0.54 (0.31-0.91)) and BCG scars (OR=0.13 (0.05-0.37)) and larger post-vaccination wheals produced more PPD reactions (OR 1.21 (95% CI 1.02-1.43)) and BCG scars (OR 1.66 (1.24-2.21)). In the multivariable analyses of BCG vaccinated children assessed at 6 months of age, monitoring of vaccination technique and type of BCG vaccine were important. This was not changed by control for other determinants, including sex, season, vaccination place, birthplace, ethnic group, low birth weight, place of residence, education and civil status of mother. We reason that vaccination technique and BCG strain are important for PPD reaction and scarring in response to BCG vaccination. Considering that these responses are associated with better infant survival, the importance of monitoring vaccination technique and of different BCG strains should be evaluated with respect to infant mortality. PMID- 15899540 TI - Polychlorinated biphenyls in polysulfide sealants--occurrence and emission from a landfill station. AB - Approximately 80,000 kg polysulfide sealant containing 10,000-18,000 kg polychlorinated biphenyls (PCB) was deposited at a Swedish municipal landfill station during 1965-1973. Investigations during 1994 showed that soil layers underneath the landfill had concentration of PCB not alarmingly high. The concentration of PCB congeners in ground water samples was elevated 4-750 times compared to a reference sample. Based on samples of ground water, leachate water, and flux chambers measuring evaporation of PCB from the landfill surface, the emission of PCB was estimated to be 1 g sigmaPCB/yr. This very low rate was attributed to the high sorptive capacity of the sealant. Compared to a reference site, the evaporation flux was elevated for the most volatile congeners, but factors 20-1400 lower than from another landfill which was contaminated with PCB in paper-pulp fibres. PMID- 15899541 TI - Order-of-magnitude increase of Hg in Norwegian peat profiles since the outset of industrial activity in Europe. AB - Peat cores from six ombrotrophic bogs at different latitudes in Norway (58 degrees N-69 degrees N) were analysed for Hg by atomic fluorescence spectrometry. In all cases a smooth decrease of Hg with depth was observed down to 15-20 cm. At greater depths Hg showed a relatively constant level of the order of 10% of that in the peat surface layer. In the surface peat Hg concentrations exhibit moderate variation with latitude. The pre-industrial levels of Hg in the peat correspond to a net annual Hg accumulation of 0.3-0.9 microgm(-2). The Hg accumulation over the last 100 years is about 15 times higher on average than the pre-industrial level. The present work supports the view that a major part of the present atmospheric Hg in the Northern Hemisphere is of anthropogenic origin. It is speculated that the comparatively high Hg contemporary accumulation rates observed at the Andoya bog on 69 degrees N may be related to the Arctic springtime depletion of Hg. PMID- 15899542 TI - Women as moral pioneers? Experiences of first trimester antenatal screening. AB - The implementation of innovative medical technologies can raise unprecedented ethical, legal and social dilemmas. This is particularly so in the area of antenatal screening, which is dominated by the language of risk and probabilities. Second trimester serum screening for Down's syndrome and neural tube defects has a well-established place in antenatal care. Increasingly, first trimester screening with biochemical and ultrasound markers is being proposed as advance on this, yielding higher detection rates of Down's syndrome at an earlier gestational age. This article explores the experiences of 14 women offered innovative first trimester screening, which takes place within the context of a detailed ultrasound scan. The study is set within the UK, where recent policy changes mean that the offer of screening for fetal anomalies, particularly Down's syndrome, will become a routine part of antenatal care and offered to all pregnant women. This paper focuses on the significance of the scan in first trimester screening, and some of the potential dilemmas for women that can result from this. It then discusses the ways in which women made their decisions about screening, in particular, their work as 'moral pioneers'. We found that the part played by the ultrasound scan in first trimester screening, particularly in relation to the higher-quality images now being obtained, has the potential to introduce new and novel ethical dilemmas for pregnant women. Although concerns have been raised about pregnant women viewing ultrasound scans as benign, many of the women reported having thought carefully through their own moral beliefs and values prior to screening. It seems that whatever other implications they may have, first trimester screening technologies will continue the tradition of pregnant women acting as 'moral pioneers' in increasingly complex settings. PMID- 15899543 TI - Does continuing oral magnesium supplementation until delivery affect labor and puerperium outcome? AB - OBJECTIVE: To determine the labor and puerperal impact of continuing oral magnesium supplementation until delivery. STUDY DESIGN: Single-center study with matched controls. In 40 pairs of healthy women with vaginally delivered singleton pregnancies, matched for maternal age, race and parity, maternal and neonatal outcome endpoints were compared in those receiving continuous oral magnesium aspartate supplementation 15-30 mmol/d for at least 4 weeks until delivery (for constipation, calf cramps, preterm contraction without cervical effacement or additional tocolytics) versus non-supplemented controls. RESULTS: In the magnesium group labor was nonsignificantly longer (stage 1: 326.0+/-187.5 min versus 276.7+/-140.8 min, P = 0.19; stage 2: 52.0+/-44.5 min versus 43.5+/-44.0 min, P = 0.40) and maximum oxytocin dose nonsignificantly higher (14.5+/-9.4 [median 12.0; n=15] versus 10.5+/-6.9 [median 7.5] mU/min, P = 0.28; n = 10). Fewer women had afterpains (12 versus 20, P=0.11), required spasmolysis (3 versus 14, P = 0.005), or could breastfeed their infants exclusively at discharge (24 versus 34, P = 0.04). CONCLUSION: Continuing oral magnesium supplementation until delivery does not significantly prolong labor or increase the oxytocin requirement, but it significantly impairs breastfeeding competence. PMID- 15899544 TI - Potential of traditional ayurvedic formulation, Triphala, as a novel anticancer drug. AB - The cytotoxic effects of aqueous extract of Triphala, an ayurvedic formulation, were investigated on human breast cancer cell line (MCF-7) and a transplantable mouse thymic lymphoma (barcl-95). The viability of treated cells was found to decrease with the increasing concentrations of Triphala. On the other hand, treatment of normal breast epithelial cells, MCF-10 F, human peripheral blood mononuclear cells, mouse liver and spleen cells, with similar concentrations of Triphala did not affect their cytotoxicity significantly. The drug treatment was found to induce apoptosis in MCF-7 and barcl-95 cells in vitro as determined by annexin-V fluorescence and proportion of apoptotic cells was found dependent on Triphala concentration. MCF-7 cells treated with Triphala when subjected to single cell gel electrophoresis, revealed a pattern of DNA damage, characteristic of apoptosis. Studies on Triphala treated MCF-7 and barcl-95 cells showed significant increase in intracellular reactive oxygen species (ROS) in a concentration dependent manner. ROS increase was, however, found to be insignificant in MCF-10 F as well as in murine spleen and liver normal cells. In vivo, direct oral feeding of Triphala to mice (40 mg/kg body weight) transplanted with barcl-95 produced significant reduction in tumor growth as evaluated by tumor volume measurement. It was also found that apoptosis was significantly higher in the excised tumor tissue of Triphala fed mice as compared to the control, suggesting the involvement of apoptosis in tumor growth reduction. These results suggest that Triphala possessed ability to induce cytotoxicity in tumor cells but spared the normal cells. The differential effect of Triphala on normal and tumor cells seems to be related to its ability to evoke differential response in intracellular ROS generation. The differential response of normal and tumor cells to Triphala in vitro and the substantial regression of transplanted tumor in mice fed with Triphala points to its potential use as an anticancer drug for clinical treatment. PMID- 15899545 TI - Effects of natural polysaccharides on the growth and peritoneal carcinomatosis of human gastric adenocarcinoma in a nude mouse model. AB - We have examined the effects of natural polysaccharides isolated from Phellinus gilvus (PG) in vitro and in vivo against gastric cancer. PG decreased cell proliferation and increased cell apoptosis in a dose-dependent manner in vitro. PG also led to a marked inhibition of tumor growth and significant decrease in the incidence of peritoneal carcinomatosis. Histological analysis of tumor confirmed dramatic increase in tumor cell apoptosis by PG, indicating reduced tumor cell proliferation. These data showed that polysaccharides isolated from PG significantly inhibited tumor growth and metastasis in an orthotopic model of human gastric adenocarcinoma, without detectable problematic adverse effects. PMID- 15899546 TI - Rapid induction of malignant tumor in Sprague-Dawley rats by injection of RK3E ras cells. AB - Several tumor animal models have been provided as a tool for developing cancer therapy. Here, we developed rapid, easy-to use, and cost-effective new rat animal model for invasion and metastasis of cancer using genetically k-ras-induced rat kidney cells (RK3E-ras). We observed tumor as early as 3 days after injection of RK3E-ras cells in subcutaneous of Sprague-Dawley rats. Tumor size and volume were increased exponentially for 2 weeks. The tail vein injected rats obtained the lethal infiltration in the lung within 2 weeks. This tumor animal model has great potential for studying cancer processes and short-term screening of variable cancer therapy strategy. PMID- 15899547 TI - Impairment of antioxidant enzymes, lipid peroxidation and 8-oxo-2'-deoxyguanosine in advanced epithelial ovarian carcinoma of a Spanish community. AB - In the present study, we describe the changes of antioxidant enzyme activities and other oxidative stress-related parameters in a mediterranean cohort of women affected with epithelial ovarian carcinoma (EOC). For that purpose, the most representative enzymatic activities, such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) and the oxidized/reduced glutathione (GSSG/GSH) ratio have been analyzed in tumor tissue biopsies and compared with the normal tissue of the same patient. As oxidation products, the levels of malondialdehyde (MDA) as an indication of lipid peroxidation, and the DNA damaged base 8-oxo-2'-deoxyguanosine (8-oxo-dG) have been also measured. Advanced EOC show reduced levels of SOD and CAT, while that of GPx is increased when compared with non-neoplastic tissue. The levels of GSH are increased giving as a result a reduction of the oxidative stress marker GSSG/GSH ratio comparing normal ovarian tissue with tumor tissue. In addition, the oxidation products MDA and 8-oxo-dG are significantly increased in tumor tissue, suggesting a shift of oxidative metabolisms towards a pro-oxidation state and potential gene instability in malignant ovary cells. The possible implication of the redox changes and DNA damage in tumor development is discussed. PMID- 15899548 TI - Anxiolytic agent, dihydrohonokiol-B, recovers amyloid beta protein-induced neurotoxicity in cultured rat hippocampal neurons. AB - The effects of anxiolytic honokiol derivative, dihydrohonokiol-B (DHH-B), on amyloid beta protein (Abeta(25-35), 10 nM)-induced changes in Cl(-)-ATPase activity, intracellular Cl- concentration ([Cl-]i) and glutamate neurotoxicity were examined in cultured rat hippocampal neurons. DHH-B (10 ng/ml) recovered Abeta-induced decrease in neuronal Cl(-)-ATPase activity without any changes in the activities of Na+/K+-ATPase and anion-insensitive Mg2+-ATPase. A GABA(C) receptor antagonist (1,2,5,6,-tetrahydropyridin-4-yl) methyl-phosphinic acid (TPMPA, 15 microM), inhibited the protective effects of DHH-B on Cl(-)-ATPase activity. DHH-B reduced Abeta-induced elevation of [Cl-]i as assayed using a Cl( )-sensitive fluorescent dye, and prevented Abeta-induced aggravation of glutamate neurotoxicity. These data suggest that DHH-B exerts the neuroprotective action against Abeta through GABA(C) receptor stimulation. PMID- 15899549 TI - GABAergic neurons express mu-opioid receptors in the ventrolateral orbital cortex of the rat. AB - Behavioral studies have indicated that GABAergic modulation is involved in the opioid-induced antinociception in the ventrolateral orbital cortex (VLO). The aim of the current study was to examine whether the GABAergic neurons in the rat VLO expressed mu-opioid receptor subtype 1 (MOR1). This study employed immunofluorescence histochemical double-staining technique and showed that a considerable amount of GABA- and MOR1-like immunoreactive neurons existed in layers II-VI in the VLO. Of these GABA-like immunoreactive neurons, 92.0% of them showed MOR1-like immunoreactivities. Similarly, 80.2% of MOR1-like immuoreactive neurons also exhibited GABA-like immunoreactivities. These results provide morphological evidence that opioid-induced antinociception in the VLO might be due to an inhibitory effect by opioid via MOR1 on GABAergic neurons, resulting in disinhibition of VLO projection neurons and leading to activation of the VLO-PAG brainstem descending pain control system to depress the nociceptive inputs at the spinal cord level. PMID- 15899550 TI - Comments on "Observation of the surface stress induced in microcantilevers by electrochemical redox processes" by F. Tian, J.H. Pei, D.L. Hedden, G.M. Brown, T. Thundat. AB - Critical remarks have been formulated in connection with the surface stress measurements reported by Tian et al. It has been shown that due to the shortcomings of the experimental procedure some conclusions of the authors are hardly acceptable. PMID- 15899551 TI - Electron optical phase-shifts by Fourier methods: analytical versus numerical calculations. AB - The theoretical framework for the computation of electromagnetic fields and electron optical phase-shifts in Fourier space has been recently applied to objects with long-range fringing fields, such as reverse-biased p-n junctions and magnetic stripe domains near a specimen edge. In addition to new analytical results, in this work, we present a critical comparison between numerical and analytical computations. The influence of explicit and implicit boundary conditions on the phase shifts and phase-contrast images is also investigated in detail. PMID- 15899552 TI - Epidemiology of Oestrus ovis (Linneo, 1761) infestation in goats in Spain. AB - This survey was conducted to determine the chronobiology and seroprevalence of nasal bot infestation (Oestrus ovis) in Spain and to identify the risk factors associated with this disease in caprine herds. A total of 1590 sera from adult goats were collected at random on 175 farms in southwestern Spain. Sera were tested by ELISA, using crude protein from second stage larvae as antigen. The mean seroprevalence was 46.04% and mean percentage of optical densities was 41.83. These data indicate a high prevalence of this parasite in the investigated areas. The serological survey revealed that goats managed at higher altitudes, at meridians latitudes and on farms with small herds had a smaller probability of infestation. Eighty goat heads, obtained from abattoirs in the central region of Spain, were collected and examined for nasal botflies from February to October 2002. O. ovis larval stages were recovered from the nasal-sinus cavities of 23 goats, reaching a prevalence of 34.94%. The mean larval burden was 3.9 larvae per infested head. No first instars were found during February and March, when the second instar reached its larger count. The third instar was observed in very small number during the whole period of study, with one peak occurring in July- August. These data show the existence of a favourable period for the development of larval instars of O. ovis in goats that starts in February and finishes in September. PMID- 15899553 TI - The predatory capability of Arthrobotrys cladodes var. macroides in the control of Haemonchus contortus infective larvae. AB - One hundred compost samples were examined for the presence of nematophagous fungi on the sheep farms of Mazanderan, province, Iran. Arthrobotrys cladodes var. macroides (IRAN 677C=CBS 143565) was isolated from 3% of the samples examined. Nematophagous activity of this fungus which was shown for the first time in this study, revealed the addition of 1000, 8000, 20000 and 100000 conidia per gram of feces of sheep reduced significantly (P<0.001) the number of Haemonchus contortus infective larvae in the feces by 41.71%, 63, 27%, 73.49% and 94.96%, respectively. These results show that A. cladodes var. macroides is a promising candidate for biological control of H. contortus. PMID- 15899554 TI - Correlates of comorbid PTSD and drinking problems among sexual assault survivors. AB - Numerous studies show that sexual assault victims are likely to develop PTSD and have a greater risk of drinking problems than nonvictims. However, little is known about what differentiates survivors with PTSD only from those with comorbid PTSD and drinking problems. In this study, a large, diverse sample of community residing women who had experienced adult sexual assault was surveyed. Logistic regression analyses were conducted to identify pre-assault, assault, and post assault factors differentiating survivors with PTSD only from those with PTSD and drinking problems. Results showed that survivors with less education, histories of other traumas, who blamed their character more for the assault, believed drinking could reduce distress, drank to cope with the assault's effects, and received negative social reactions were more likely to have comorbid PTSD and drinking problems than those with PTSD only. Implications for future research and treatment with sexual assault victims are discussed. PMID- 15899555 TI - Elevated levels of soluble interleukin-2 receptor in serum of patients with hematological or non-hematological malignancies. AB - We measured the levels of soluble interleukin-2 receptor (sIL-2R) in the serum of 53 patients with hematological malignancies (14 with acute leukemia, 10 myeloproliferative disorders, 21 non-Hodgkin's lymphoma and 8 multiple myeloma), 40 with non-hematological solid malignancies (24 with lung cancer, 11 digestive system cancer and 5 other cancers) and 95 healthy subjects as a control using an enzyme-linked immunosorbent assay. Both hematological and non-hematological neoplasms showed significantly higher sIL-2R levels than normal subjects (p<0.01). However, compared to solid malignancies, hematological neoplasms displayed a wide range of sIL-2R levels and extremely elevated values of sIL-2R were seen in certain cases. These results seem to suggest that sIL-2R levels may serve as one of non-invasive markers of differential diagnosis for patients with bulky mass lesions between hematological and solid malignancy. PMID- 15899556 TI - Increasing treatment options for cannabis dependence: a review of potential pharmacotherapies. AB - Despite the fact that rates of cannabis dependence have increased substantially over the past several years, there are no medications approved for the treatment of cannabis dependence. This paper reviews data from recent research on cannabinoids that may be relevant for the development of pharmacotherapies for cannabis dependence. Included in the discussion are findings from studies that have assessed the ability of medications to ameliorate cannabis-related abstinence symptoms in laboratory animals and human research participants. Data from studies that have investigated the effects of pharmacological agents on cannabis self-administration are also reviewed because these data may provide information critical for informing relapse prevention medication development efforts. The majority of published studies evaluating cannabis pharmacotherapies have focused on decreasing withdrawal symptoms: a growing number of medications reduce symptoms in laboratory animals, but the majority of these medications have not been tested in humans. Fewer studies have assessed the effects of potential cannabis treatment medications on cannabinoid-related reinforcing effects. In laboratory animals, only the CB1 cannabinoid antagonist rimonabant has shown promise. In humans, this medication has not been tested on cannabis reinforcing effects. To date, no medication has been shown to alter cannabis self administration by humans. PMID- 15899557 TI - Reducing hospital presentations for opioid overdose in patients treated with sustained release naltrexone implants. AB - BACKGROUND: Non-fatal overdoses represent a significant morbidity for regular heroin users. Naltrexone is an opioid antagonist capable of blocking the effects of heroin, thereby preventing accidental overdose. However, treatment with oral naltrexone is often associated with non-compliance. An alternative is the use of a sustained release preparation of naltrexone. The aim of this study was to assess the change in number of opioid and other drug overdoses in a large cohort of heroin dependent persons (n=361; 218 males) before and after treatment with a sustained release naltrexone implant. A sub-group of this cohort (n=146; 83 males) had previously received treatment with oral naltrexone, which also allowed a comparison of overdoses pre- and post-oral and also post-implant treatments. METHOD: We used a pre-post design, with data prospectively collected via the West Australian Health Services Research Linked Database, and the Emergency Department Information System. Participants were treated under the Australian Therapeutic Goods Administration's special access guidelines. RESULTS: Most (336, 93%) of the cohort was in one or both databases. We identified 21 opioid overdoses involving 20 persons in the 6 months pre-treatment that required emergency department presentation or hospital admission: none were observed in the 6 months post treatment. This is consistent with the existing pharmacokinetic data on this implant, which indicates maintenance of blood naltrexone levels at or above 2 ng/ml for approximately 6 months. A reduced number of opioid overdoses were also observed 7-12 months post-implant. The study found a significant increase in sedative "overdoses", some of which occurred in the 10 days following implant treatment and were likely associated with opioid withdrawal and/or implant treatment. For those previously treated with oral naltrexone, more opioid overdoses occurred in both the 6-months prior to and after oral compared to the 6 months post-implant treatment. CONCLUSIONS: The findings support the clinical efficacy of this sustained release naltrexone implant in preventing opioid overdose. However, given the high prevalence of poly-substance use among dependent heroin users, programs offering this type of treatment should also focus on preventing, detecting and managing poly-substance use. PMID- 15899558 TI - Dual-subtype vaccine (Fel-O-Vax FIV) protects cats against contact challenge with heterologous subtype B FIV infected cats. AB - Fel-O-Vax FIV is a dual-subtype vaccine consisting of inactivated whole viruses of subtype A (Petaluma strain) and subtype D (Shizuoka strain). The efficacy of this vaccine against heterologous subtype A strain challenge was demonstrated, but it is unclear whether the result reflects efficacy in the field. In this study, we evaluated the efficacy of this vaccine against contact challenge by exposing both vaccinated and unvaccinated control animals with cats infected with Aomori-2 strain belonging to subtype B, a subtype prevalent in many regions of the world. Nineteen specific-pathogen-free (SPF) cats were divided into a vaccinated group (six cats), an unvaccinated control group (eight cats), and a challenge group (five cats), and maintained in the same room. Cats were monitored for FIV proviral DNA by nested PCR and for FIV-specific antibody levels by ELISA. After 1 year of commingling, each cat in the vaccinated group was given a booster dose. In addition, the original challenge group was removed and replaced with another challenge group of SPF cats, which were inoculated with the Aomori-2 strain. FIV infection was confirmed in four of the eight animals in the unvaccinated control group by the 29th week in the second year of commingling. In contrast, all of the animals were negative in the vaccinated group. These findings confirmed the efficacy of this vaccine against heterologous stains classified as subtype B, and suggested that the vaccine exhibits broad efficacy against genetically diverse FIV. PMID- 15899559 TI - Protamine sulfate protects exogenous DNA against nuclease degradation but is unable to improve the efficiency of bovine sperm mediated transgenesis. AB - The present study investigated whether protamine sulfate can be used to improve the efficiency of bovine sperm mediated transgenesis (SMT) by protecting the plasmid pCX-EGFP against nuclease activity. A high proportion (31%) of bovine spermatozoa transfected with the plasmid pCX-EGFP maintain their motility. Using an in vitro assay, protamine sulfate protected the plasmid against degradation by DNase I. However, upon transfecting spermatozoa, the plasmid remained intact regardless of whether it was complexed to protamine sulfate. When in vitro fertilisation (IVF) was undertaken using transfected sperm, 14.6 and 10.2% of blastocysts derived from pCX-EGFP only and pCX-EGFP-protamine transfected sperm, respectively, were PCR positive for the plasmid. In conclusion, using spermatozoa transfected with either pCX-EGFP or pCX-EGFP-protamine complexes, produced PCR positive blastocysts after SMT. However, the use of protamine sulfate does not improve the efficiency of SMT suggesting that factors other than nuclease activity could be limiting. PMID- 15899560 TI - Development to the blastocyst stage of immature pig oocytes arrested before the metaphase-II stage and fertilized in vitro. AB - In vitro fertilization (IVF) and embryonic development of mature and meiotically arrested porcine oocytes were compared in the present study. After in vitro maturation (IVM) of cumulus-oocyte complexes for 48 h, 75.4% of them extruded a visible polar body (PB). Most of the oocytes with a first polar body (PB+ group) were at the metaphase-II (M-II) stage (91.4%). Most of the oocytes without a visible polar body (PB- group) appeared to be arrested at the germinal vesicle (GV) (41.6%) and metaphase-I (M-I) (34.0%) stages. After IVF of oocytes (day of IVF=Day 0), there was no difference between PB+ and PB- groups in rates of sperm penetration, mono-spermy, however oocyte activation rate after penetration was greater in the PB+ than in the PB- group (P<0.05). On Day 2, there was no difference between rates of embryos cleaved at the 2-4 cell stages in PB+ and PB- groups (42.1+/-48.8% and 33.6+/-2.1%, respectively). On Day 4, the rate of PB+ embryos developing beyond the 4-cell stage was greater than that of PB- embryos (P<0.05, 31.7+/-3.9% and 14.1+/-1.5%, respectively), and PB+ embryos had more cells than the PB- embryos (P<0.05, 8.3+/-0.4 and 6.0+/-0.8 cells, respectively). On Day 6, a greater proportion of PB+ embryos developed to the blastocyst stage than did PB- embryos (P<0.05, 34.6+/-2.4% and 20.7+/-2.8%, respectively). However, when the GV oocytes of the PB- group were not included in recalculations, there was no difference in blastocyst rates between M-I arrested and M-II oocytes (35.3 and 34.6%, respectively). The number of blastomere nuclei in embryos obtained from the PB+ group (52.0+/-2.5) was greater than that from the PB- group (P<0.05, 29.1+/-2.8). The proportion of degenerated parts in the blastocysts, as determined by morphological appearance, was the same in the PB+ and PB- groups. Although the quality of PB+ embryos was enhanced as compared with that of the PB- group, the proportion of inner cell mass and trophectoderm cells in PB+ and PB- blastocysts did not differ (1:1.9 and 1:2.2, respectively). Chromosome analysis revealed that PB+ blastocysts had more diploidy (P<0.05, 69.7%) than did PB- blastocysts (44.0%), whereas PB- blastocysts had more triploid cells (P<0.05, 34.0%) than did PB+ oocytes (8.4%). These results indicate that pig oocytes arrested before the M-II stage (M-I oocytes) undergo cytoplasmic maturation during maturation culture and have the same ability to develop to blastocysts after IVF as M-II oocytes, but some of them resulted in degeneration or delayed development with poor embryo quality. PMID- 15899561 TI - Nanoencapsulation of a crystalline drug. AB - The aim of this work was to assess the influence of various formulation parameters on the incorporation of a poorly water-soluble crystalline drug into nanoparticles. For this purpose, the influence of the polymer (polylactic acid, polysebacic acid terminated with lithocholic acid, and polysebacic acid-co lithocholic acid) as well as the effect of the dispersion medium (aqueous phases at different temperatures, saline medium and ethanol) on the encapsulation was investigated. 3H-labelled drug was used in order to determine the loading efficiency by liquid scintillation counting. The solubility of the drug in the various polymer materials was assessed by differential scanning calorimetry (DSC). The solubility of the drug in the different dispersion media was then determined by gas chromatographic-mass spectrometric measurements. The highest loading ratios were obtained using poly (lactic acid) (PLA). However, the drug solubility in the polymers, determined by DSC analysis, cannot be considered as predictive for encapsulation efficiency. The study of the influence of the liquid outer phase showed that the encapsulation efficiency increased when the drug solubility in the dispersion medium (before acetone evaporation) decreased. These experiments made it possible to propose a mechanism to account for the leakage of the crystalline drug during the nanoprecipitation process. So, when acetone is eliminated by evaporation, the drug solubility in the dispersion medium decreases, leading to the formation of crystals. During nanoparticles storage, the crystals continue to grow, the nanoparticles serving as drug reservoirs. These findings highlight the importance of using a polymer with a specific affinity for the drug, and a dispersion medium with the lowest drug solubility to achieve an efficient encapsulation of a crystalline drug. PMID- 15899562 TI - Anti-ulcerogenic activity of xanthanolide sesquiterpenes from Xanthium cavanillesii in rats. AB - The preventive effect of natural xanthanolides as well as a series of synthetic derivatives on ulcer formation induced by absolute ethanol in rats was examined. Among the compounds tested, xanthatin gave the strongest protective activity. The inhibitory action exerted by this molecule on the lesions induced by 0.6N HCl and 0.2N NaOH was highly significant, reducing ulceration in the range of 58-96% at a dose from 12.5 to 100mg/kg. These results appear to confirm that the presence of a non-hindered alpha,beta-unsaturated carbonyl group seems to be an essential structural requirement for the gastric cytoprotective activity of these compounds. In order to explore this possibility, a theoretical conformational analysis was performed. We suggest that the mechanism of protection would involve, at least in part, a nucleophylic attack of the sulfhydryl group from the biological molecules present in the gastric mucosa to electrophylic carbons accessible in suitable Michael acceptors. PMID- 15899563 TI - Effects of ethanol and water extracts of propolis (bee glue) on acute inflammatory animal models. AB - The anti-inflammatory effects of ethanol (EEP) and water (WSD) extracts in ICR mice and Wistar rats were analyzed. Both WSD and EEP exhibited significant anti inflammatory effects in animal models with respect to thoracic capillary vessel leakage in mice, carrageenan-induced oedema, carrageenan-induced pleurisy, acute lung damage in rats. The mechanisms for the anti-inflammatory effects probably involve decreasing prostaglandin-E(2) (PGE(2)) and nitric oxide (NO) levels. In rats with Freund's complete adjuvant (FCA) induced arthritis, propolis extracts significantly inhibited the increase of interleukin-6 (IL-6) in inflamed tissues, but had no significant effect on levels of interleukin-2 (IL-2) and interferon gamma (IFN-gamma). The results are consistent with the interpretation that EEP and WSD may exert these effects by inhibiting the activation and differentiation of mononuclear macrophages. PMID- 15899564 TI - Drug screening and confirmation by GC-MS: comparison of EMIT II and Online KIMS against 10 drugs between US and England laboratories. AB - Drug screening through urinalysis is a widely accepted tool for rapid detection of potential drug use at a relatively low cost. It is, therefore, a potentially useful method for detecting and monitoring drug use in a variety of contexts such as the criminal justice system, pre-employment screening and a variety of treatment centers. This article explores the efficacy of two commercially available drug-screening assays: Online KIMS assay (Roche) and EMIT II assays. First, we evaluate the sensitivity and specificity of two immunoassays. A total of 738 urine samples were collected among adult arrestee populations from Chicago, New Orleans and Seattle through the Arrestee Drug Abuse Monitoring (ADAM) program. Partial samples were split within one laboratory and analyzed by both enzymes multiplied immunoassay technique (EMIT) II and kinetic interaction of microparticle in solution (KIMS) assays for a 10-drug panel (amphetamine, barbiturates, benzodiazepines, marijuana, cocaine, methadone, methaqualone, opiate, phencyclidine and propoxyphene). Gas chromatography-mass spectrometry (GC MS) was used as a confirmation method for all positives from either EMIT II or KIMS for all experiments. Second, the paper examines whether using different testing laboratories plays a role in the final results. The same experiments were repeated at two different testing locations: one in California and one in London and England. Third, the paper studies whether drug testing results vary between two laboratories when each of them had used their own routine screening method: the Forensic Science Service (FSS) at Birmingham, United Kingdom with KIMS assay and Medscreen Limited at London, United Kingdom with EMIT II. In summary, both EMIT II and KIMS assays generate fairly consistent results. The concordance rate against each of the 10 drugs tested is relatively high (97.4-100%). The discrepancies, in most cases, occurred at drug concentrations near the cut-off levels. There were more discrepant results between two laboratories compared to when specimens were analyzed at the same laboratory using two different assays. PMID- 15899565 TI - Oral fluid collection: the neglected variable in oral fluid testing. AB - The potential to use oral fluid as a drug-testing specimen has been the subject of considerable scientific interest. The ease with which specimens can be collected and the potential for oral fluid (OF) drug concentrations to reflect blood-drug concentrations make it a potentially valuable specimen in clinical as well as forensic settings. However, the possible effects of the OF collection process on drug detection and quantification has often been over looked. Several studies have documented that drug-contamination of the oral cavity may skew oral fluid/blood drug ratios and confound interpretation when drugs are smoked, insufflated or ingested orally. OF pH is predicted to have an effect on the concentration of drugs in OF. However, in a controlled clinical study, the effect of pH was less than that of collection technique. Mean codeine OF concentrations in specimens collected a non-stimulating control method were 3.6 times higher than those in OF collected after acidic stimulation. Mean codeine concentrations were 50% lower than control using mechanical stimulation and 77% of control using commercial collection devices. Several factors should be considered if a commercial OF collection device is used. In vitro collection experiments demonstrated that the mean collection volume varied between devices from 0.82 to 1.86 mL. The percentage of the collected volume that could be recovered from the device varied from 18% to 83%. In vitro experiments demonstrated considerable variation in the recovery of amphetamines (16-59%), opiates (33-50%), cocaine and benzoylecgonine (61-97%), carboxy-THC (0-53%) and PCP (9-56%). Less variation in collection volume, volume recovered and drug recovery was observed intra-device. The THC stability was evaluated in a common commercial collection protocol. Samples in the collection buffer were relatively stable for 6 weeks when stored frozen. However, stability was marginal under refrigerated conditions and poor at room temperature. Very little has been published on the efficacy of using IgG concentration, or any other endogenous marker, as a measure of OF specimen validity. Preliminary rinsing experiments with moderate (50 mL and 2 x 50 mL) volumes of water did not reduce the OF IgG concentration below proposed specimen validity criteria. In summary, obvious and more subtle variables in the OF collection may have pronounced effects on OF-drug concentrations. This has rarely been acknowledged in the literature, but should to be considered in OF drug testing, interpretation of OF-drug results and future research studies. PMID- 15899566 TI - Prolonged hypothermia protects neonatal rat brain against hypoxic-ischemia by reducing both apoptosis and necrosis. AB - Although hypothermia is an effective treatment for perinatal cerebral hypoxic ischemic (HI) injury, it remains unclear how long and how deep we need to maintain hypothermia to obtain maximum neuroprotection. We examined effects of prolonged hypothermia on HI immature rat brain and its protective mechanisms using the Rice-Vannucci model. Immediately after the end of hypoxic exposure, the pups divided into a hypothermia group (30 degrees C) and a normothermia one (37 degrees C). Rectal temperature was maintained until they were sacrificed at each time point before 72h post HI. Prolonged hypothermia significantly reduced macroscopic brain injury compared with normothermia group. Quantitative analysis of cell death using H&E-stained sections revealed the number of both apoptotic and necrotic cells was significantly reduced by hypothermia after 24h post HI. Hypothermia seemed to decrease the number of TUNEL-positive cells. Immunohistochemistry and Western blot showed that prolonged hypothermia suppressed cytochrome c release from mitochondria to cytosol and activation of both caspase-3 and calpain in cortex, hippocampus, thalamus and striatum throughout the experiment. These results showed that prolonged hypothermia significantly reduced neonatal brain injury even when it was started after HI insult. Our results suggest that prolonged hypothermia protects neonatal brain after HI by reducing both apoptosis and necrosis. PMID- 15899567 TI - A stability indicating LC method for zolmitriptan. AB - A gradient, reversed-phase liquid chromatographic (RP-LC) assay method was developed for the quantitative determination of zolmitriptan, used to treat severe migraine headaches. The developed method is also applicable for the related substances determination in bulk drugs. The chromatographic separation was achieved on a Waters X Terra RP18, 250 mm x 4.6 mm, 5 microm column. The gradient LC method employs solutions A and B as mobile phase. The solution A contains a mixture of phosphate buffer pH 9.85:methanol:acetonitrile (70:20:10, v/v/v) and solution B contains a mixture of phosphate buffer, pH 9.85:acetonitrile (30:70). The flow rate was 1.0 ml/min and the detection wavelength was 225 nm. In the developed HPLC method, the resolution between zolmitriptan and its potential impurities, namely Imp-1, Imp-2 and Imp-3 was found to be greater than 3. The drug was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. Considerable degradation was found to occur in alkaline medium and oxidative stress conditions. Degradation product formed during base hydrolysis was found to be Imp 3. The stress samples were assayed against a qualified reference standard and the mass balance was found close to 99.5%. The developed RP-LC method was validated with respect to linearity, accuracy, precision and robustness. PMID- 15899568 TI - Simultaneous determination of naproxen, ketoprofen and phenol red in samples from rat intestinal permeability studies: HPLC method development and validation. AB - A simple reversed-phase high performance liquid chromatographic method with UV detection at 270 nm was developed for simultaneous quantitation of ketoprofen and naproxen sodium along with phenol red as a non-absorbable marker for in situ permeability studies. The mobile phase was a mixture of 20% methanol, 28% of acetonitrile, 52% water and 0.4 ml triethylamine (adjusted to pH 3.2 using orthophosphoric acid). Analysis was run at a flow of 1.5 ml/min with a 20 min run time. The calibration curves were linear for all three compounds (r>0.999) across the concentration range of 15.6-250 microg/ml with a limit of quantitation of 0.3, 0.25 and 0.2 ng/ml for naproxen, ketoprofen and phenol red, respectively. The coefficient of variation for intra-assay and inter-assay precision was less than or equal to 5.3% and the accuracy was between 95.36 and 101.6%. Using the SPIP technique and the suggested HPLC method for sample analysis, the mean values of 1.17e(-4) (+/-0.28) cm/s and 0.97e(-4) (+/-0.2) cm/s were obtained for naproxen and ketoprofen, respectively. PMID- 15899569 TI - Determination of roxithromycin in rat lung tissue by liquid chromatography-mass spectrometry. AB - A liquid chromatography-mass spectrometry (LC-MS) method for the determination of roxithromycin in rat lung tissue is described. Liquid-liquid extraction was adopted for sample preparation with recoveries from 72.5 to 76.9% at levels of 0.1, 5.0 and 20.0 microg/ml. Chromatographic separation was performed on a C18 column using a mixture of methanol, water and formic acid (80:20:1, v/v/v) as mobile phase delivered at a flow rate of 0.5 ml/min. Positive selected ion monitoring (SIM) mode was used for the quantification of roxithromycin at m/z 837.7 and clarithromycin (internal standard) at m/z 748.7. The linearity was obtained over the concentration range of 0.05-20.0 microg/ml and the lower limit of quantification was 0.05 microg/ml. For each QC level of roxithromycin, the intra- and inter-day precisions relative standard deviation (R.S.D.) were less than 4.1 and 7.5%, respectively, and accuracy (RE) was +/-10.0%. The proposed LC MS method has been successfully used for the determination of roxithromycin in rat lung tissue after oral administration of roxithromycin formulations to 44 SD rats. The present study demonstrates that the concentration of roxithromycin in rat lung tissues can be significantly increased by ambroxol when they are formulated in combination. PMID- 15899570 TI - Analysis of low level radioactive metabolites in biological fluids using high performance liquid chromatography with microplate scintillation counting: method validation and application. AB - TopCount, a microplate scintillation counter (MSC), has been recently employed as an off-line liquid radiochromatographic detector for radioactive metabolite profile analysis. The present study was undertaken to validate TopCount for metabolite profiling with respect to sensitivity, accuracy, precision and radioactivity recovery. Matrix effects of various human samples on TopCount performance and capability of MSC for volatile metabolite analysis were also investigated. TopCount had a limit of detection (LOD) of 5 DPM and a limit of quantification (LOQ) of 15 DPM for [(14)C]-labeled compounds at a 10min counting time. It was two-fold more sensitive than a liquid scintillation counter (LSC), and 50-100-fold more sensitive than a radioactivity flow detector (RFD). TopCount had comparable accuracy and precision to RFD, and comparable precision to LSC for determining relative abundance of metabolites. Human liver microsome incubation (up to 1 mL), plasma (up to 1 mL), urine (up to 2 mL) and feces (up to 50mg) had no significant quenching effects on TopCount performance. Benzoic acid, a volatile metabolite, was detected by TopCount, but not by Microbeta counter after microplates were dried under vacuum. Radioactivity recovery in HPLC-MSC analysis was reliably determined using an LSC-based method. Examples of using HPLC-MSC for analysis of low levels of radioactive metabolites are presented, including determination of plasma metabolite profile, in vitro reactive metabolites trapped by [(3)H]glutathione, and metabolite concentrations in an enzyme kinetic experiment. The data from this study strongly suggest that HPLC in combination with TopCount is a viable alternative analytical tool for detection and quantification of low levels of radioactive metabolites in biological fluids. PMID- 15899571 TI - Development and validation of an LC-UV method for the quantification and purity determination of the novel anticancer agent C1311 and its pharmaceutical dosage form. AB - C1311 (5-[[2-(diethylamino)ethyl]amino]-8-hydroxyimidazo [4,5,1-de]-acridin-6-one dihydrochloride trihydrate) is the lead compound from the group of imidazoacridinones, a novel group of rationally designed anticancer agents. The pharmaceutical development of C1311 necessitated the availability of an assay for the quantification and purity determination of C1311 active pharmaceutical ingredient (API) and its pharmaceutical dosage form. A reversed-phase liquid chromatographic method (RP-LC) with ultraviolet (UV) detection was developed, consisting of separation on a C18 column with phosphate buffer (60 mM; pH 3 with 1 M citric acid)-acetonitrile-triethylamine (83:17:0.05, v/v/v) as the mobile phase and UV-detection at 280 nm. The method was found to be linear over a concentration range of 2.50-100 microg/mL, precise and accurate. Accelerated stress testing showed degradation products, which were well separated from the parent compound, confirming its stability-indicating capacity. Moreover, the use of LC-MS and on-line photo diode array detection enabled us to propose structures for four degradation products. Two of these products were also found as impurities in the API and more abundantly in an impure lot of API. PMID- 15899572 TI - Differentiation of bovine from porcine gelatines using polyclonal anti-peptide antibodies in indirect and competitive indirect ELISA. AB - Gelatine is a collagen derivative obtained from bones and hides/skin mainly from bovine and pigs. As a consequence of the outbreak of bovine spongiform encephalopathy (BSE), the use of bovine gelatine in feed, food and pharmaceutical products has been restricted by regulatory authorities. However, no method was presently available for its specific detection. The large similarity in amino acid sequences of collagens from different species make their immunochemical differentiation difficult when using polyclonal antibodies raised against the whole molecule [A. Venien, D. Levieux, J. Immunoassay Immunochem., in press]. To obtain bovine-specific antibodies, we immunized rabbits against putative species specific sequences of the bovine collagen alpha 1(I) chain. Using these antibodies, an indirect ELISA was developed to allow a quick and easy differentiation between bovine and porcine gelatines. Moreover, a competitive indirect ELISA was found suitable to detect bovine gelatine in porcine gelatine purchased from laboratory chemicals suppliers down to a dilution of 2-4 parts per 1000 with CVs ranging from 5.7 to 7.7%. When testing mixtures of the largest possible range of industrial batches of bovine and porcine gelatines (skin/hides or bones origin, acid or alkaline processes, high or low Bloom) the detection limit was down to a dilution of 8 parts per 100 bovine gelatine in porcine gelatine. These ELISAs could be routinely used by pharmaceutical and food manufacturers to secure their supply chain. PMID- 15899573 TI - Growing E. coli to high cell density--a historical perspective on method development. AB - E. coli is the major bacterial platform for expressing simple heterologous proteins. Growing E. coli to high densities has been the subject of numerous studies since the early 1970s, exploring the limits of bacterial culture density in order to achieve maximum productivity. Research strategies were focused on improving the cultivation techniques, manipulating the bacteria's physiology or both. As a result, batch, fed batch and dialysis fermentation techniques had been developed. These growth strategies, together with optimization of media composition and the application of molecular biology methods, made it possible to grow E. coli to cell densities of up to 190 g/l (dry weight), while avoiding media precipitation and preventing acetate accumulation. Additional research on the effects of heterologous protein biosynthesis on signal transduction, proteolysis and post transcription events in E. coli may improve its productivity. PMID- 15899574 TI - Iron acquisition by Actinobacillus suis: identification and characterization of a single-component haemoglobin receptor and encoding gene. AB - Actinobacillus suis is an important swine pathogen. As with other pathogens, the ability of A. suis to acquire iron within the host is crucial for virulence. Here, we investigated the ability of seven strains of A. suis to acquire iron from haemoglobins. In growth assays, all strains could use porcine, bovine and human haemoglobins as iron sources for growth. Using solid phase binding assays, membranes derived from all strains, grown under iron-restricted conditions, were shown to bind all three haemoglobins. Competition binding assays indicated that these haemoglobins were bound by the same receptor and an affinity procedure allowed the isolation and identification of an iron-repressible, haemoglobin binding polypeptide (approximately 105 kDa) from all strains. Nucleotide sequence analyses revealed that A. suis possesses a gene (hgbA) that encodes a homologue of the Actinobacillus pleuropneumoniae haemoglobin-binding protein, HgbA. hgbA, encoding a mature protein of 105 kDa, was shown to be preceded by a hugZ homologue; putative promoter sequences and a putative Fur box were located upstream of hugZ and RT-PCR revealed that hugZ and hgbA are co-transcribed and iron-repressible. It is concluded that the acquisition of haemoglobin-bound iron by A. suis involves a single-component receptor that is up-regulated in response to iron restriction. PMID- 15899575 TI - Bacteria-associated fibronectin does not enhance Chlamydia trachomatis infectivity in vitro. AB - The molecules responsible for mediating attachment of pathogenic bacteria to their host cells are essential determinants of the pathogen's success. Staphylococci, streptococci, and gonococci are known to utilize fibronectin to mediate adhesion and/or entry into eukaryotic cells. It has been shown that Chlamydia trachomatis binds fibronectin via a heparan sulfate-lyase sensitive molecule on the surface of the bacteria. Because heparan sulfate-lyase treated Chlamydia are markedly reduced in infectivity, we hypothesized that fibronectin might be acting as an intermediate to form a molecular bridge between heparan sulfate-like molecules on the surface of Chlamydia and integrins on the host cell. To test this question, fibronectin-deficient C. trachomatis were used in a series of infectivity assays whereby exogenous fibronectin was added to the inoculum. At concentrations up to 100 ug/ml fibronectin did not modulate C. trachomatis infectivity. However, at high fibronectin concentrations (1 mg/ml) bacterial infectivity was reduced. These data suggest that, unlike several other microbial pathogens, C. trachomatis does not utilize fibronectin to mediate infectivity in vitro. PMID- 15899576 TI - The integrated project ReProTect: a novel approach in reproductive toxicity hazard assessment. AB - Validated alternative test methods are urgently needed for safety testing of drugs, chemicals and cosmetics. Whereas some animal tests for topical toxicity have been successfully replaced by alternative methods, systemic toxicity testing requires new test strategies in order to achieve an adequate safety level for the consumer. Substantial numbers of animals are required for the current in vivo assays for drugs, chemicals and cosmetics and a broad range of pioneering alternative methods were already developed. These prerequisites motivate the development of a tiered testing strategy based on alternative tests for reproductive toxicity hazard. In the Integrated Project ReProTect, a consortium set up by the European Centre for the Validation of Alternative Methods (ECVAM) takes the lead to manage the development of a testing strategy in the area of reproductive toxicity. The reproductive cycle can be broken down into well defined sub-elements, namely male and female fertility, implantation and pre/postnatal development. In this project, in vitro, in silico and sensor technologies will be developed, leading to testing strategies, that shall be implemented and disseminated. PMID- 15899577 TI - Re-distribution of phospholipase C gamma 2 in macrophage precursors is mediated by the actin cytoskeleton under the control of the Src kinases. AB - Macrophage colony-stimulating factor (M-CSF) is a growth factor that is known to trigger several signalling pathways through receptor tyrosine kinase activation. We investigated the specific requirements for the activation of phospholipase C gamma 2 (PLC-gamma2) during the differentiation of mouse bone marrow-derived macrophage precursors. M-CSF stimulation induced rapid PLC-gamma2 translocation and phosphorylation from the cytosolic compartment to the cell periphery. Both events were dependent on cytoskeleton integrity and Src kinase activity, but only PLC-gamma2 phosphorylation did not require PI3-kinase activity. Biochemical experiments as well as confocal microscopy analyses indicate that the translocation of PLC-gamma2 is mediated by the direct association of this protein with the actin cytoskeleton. Using GST-fusion proteins containing various deletions of the PLC-gamma2 Src homology region, it was found that PLC-gamma2 binds to F-actin via its SH2 domains, a feature that has equally been found in a co-sedimentation assay. This association, which is increased during actin reorganisation and disrupted by cytoskeleton inhibitors, seems to be a primary means to recruit this enzyme to the cell periphery. These results indicate that, upon M-CSF stimulation, PLC-gamma2 cellular localisation and phosphorylation are strongly dependent on cytoskeleton architecture of the macrophage precursor as well as the PI3-kinase and the Src kinases. PMID- 15899578 TI - Correction of estimation of allelic frequency in Galderisi et al. study. PMID- 15899579 TI - A detailed method for isolation and purification of human cutaneous mast cells. PMID- 15899580 TI - Cyclophosphamide decreases the number, percentage and the function of CD25+ CD4+ regulatory T cells, which suppress induction of contact hypersensitivity. AB - BACKGROUND: It is well known that cyclophosphamide (Cy) treatment before sensitization paradoxically enhances rather than suppresses contact hypersensitivity (CH) reactions. In fact, Cy-treated mice developed a significant (p < 0.05) increase of the CH reactions to 2,4,6-trinitro-1-chrolobenzene (TNCB) in comparison with untreated mice. OBJECTIVE: In order to examine whether the target cells of Cy in the immuno-augmentative effect are CD25(+) CD4(+) regulatory T cells or not, we investigated effect of Cy treatment on CD25(+) CD4(+) T cells. METHOD: We examined Cy-treated CD25(+) CD4(+) T cells by flow cytometer and by inhibition assay on proliferation of CD25(-) CD4(+) T cells. RESULTS: Cy treatment remarkably reduced the number and percentage of CD25(+) CD4(+) T cells in the spleen and lymph nodes 3 and 5 days later. Moreover, CD25(+) CD4(+) T cells taken from the Cy-treated mice 3 days later showed the lower suppressive activity on proliferation of CD25(-) CD4(+) T cells, as compared to that from the untreated mice. Furthermore, transfer of CD25(+) CD4(+) T cells from untreated mice resulted in a significant decrease (p < 0.05) of the CH reactions enhanced by Cy treatment. CONCLUSION: These results indicate that enhancement of the CH reactions to TNCB by Cy treatment is attributed to the decrease in the number, percentage and the function of CD25(+) CD4(+) regulatory T cells. PMID- 15899581 TI - 1H/31P distance determination by solid state NMR in multiple-spin systems. AB - The results of two techniques of dipolar recoupling, REDOR and CPMAS, are compared in the case of a coupled multiple-spin system. A fundamentally different behavior is observed for these two techniques. In REDOR, the terms associated with each interaction S-I(k) commute with each other and no truncation takes place so that each addition of spin I(k) causes a splitting with its dipolar frequency. In CPMAS, the flip-flop terms of the dipolar Hamiltonian do not commute with the dominant term from the strongly coupled spin pair so that the weak coupling terms from the neighboring spin I(k) are effectively truncated by the dominant pair interaction. Spin dynamics calculations are in agreement with the experimental data in a cubane shaped cluster. PMID- 15899582 TI - Solid-state NMR characterisation of the thermal transformation of a Hungarian white illite. AB - (1)H, (27)Al, (29)Si and (39)K solid-state NMR are reported from a Hungarian illite 2:1 clay for samples heated up 1600 degrees C. This single-phase sample has a small amount of aluminium substitution in the silica layer and very low iron-content ( approximately 0.4wt%). Thermal analysis shows several events that can be related to features in the NMR spectra, and hence changes in the atomic scale structure. As dehydroxylation occurs there is increasing AlO(4) and AlO(5) contents. The silica and gibbsite layers become increasingly separated as the dehydroxylation progresses. Between 900 and 1000 degrees C the silica layer forms a potassium aluminosilicate glass. The gibbsite-layer forms spinel/gamma Al(2)O(3) and some aluminium-rich mullite. Then on heating to 1600 degrees C changes in the (29)Si and (27)Al MAS NMR spectra are consistent with the aluminosilicate glass increasing its aluminium-content, the amount of mullite increasing probably with its silicon-content also increasing, and some alpha Al(2)O(3) forming. PMID- 15899583 TI - Effect of eicosapentaenoic acid (EPA) on expression of a lipid mobilizing factor in adipose tissue in cancer cachexia. AB - Adipose tissue of mice bearing a cachexia-inducing murine tumour (MAC16) shows increased expression of zinc-alpha(2)-glycoprotein (ZAG), a lipolytic factor thought to be responsible for the increased lipolysis. The anti-cachectic agent eicosapentaenoic acid (EPA) (0.5 g/kg) attenuated the loss of body weight in mice bearing the MAC16 tumour, and this was accompanied by downregulation of ZAG expression in both white and brown adipose tissue, as determined by Western blotting. Glucocorticoids may be responsible for the increased ZAG expression in adipose tissue. Dexamethasone (1.68 microM) stimulated lipolysis in 3T3-L1 adipocytes, and this effect was attenuated by EPA (50 microM). In addition the lipolytic action of dexamethasone was attenuated by anti-ZAG antibody, suggesting that the induction of lipolysis was mediated through an increase in ZAG expression. This was confirmed by Western blotting, which showed that dexamethasone (1.68 microM) induced a two-fold increase in ZAG expression in both cells and media, and that this was attenuated by EPA (50 microM). These results suggest that EPA may preserve adipose tissue in cachectic mice by downregulation of ZAG expression through interference with glucocorticoid signalling. PMID- 15899584 TI - Impact of coenzyme regeneration on the performance of an enzyme-based optical biosensor: a computational study. AB - A mathematical model of a reagent-less optical sensing scheme composed of an enzymatic reaction coupled to light-controlled photochemical coenzyme regeneration is described. The model is based on previous experimental work describing the regeneration of NADPH from NADP(+) by excited state thionine coupled to the oxidation of isocitrate by isocitrate dehydrogenase. The system is capable of repeated isocitrate measurements without the addition of exogenous coenzyme. The model is simulated using numerical integration to determine the effect of regeneration on the sensor sensitivity, response time and maximum sample throughput rate. Prediction of these effects without a model is difficult due to activation and inhibition of the dehydrogenase by both forms of the coenzyme. The regeneration parameters, including thionine concentration and thionine excitation pattern, are varied to determine optimal sensor conditions to maximize performance. A periodic regeneration approach is found to be superior to a continuous regeneration approach as the former maximizes sensitivity and minimizes response time in most cases. In addition periodic regeneration results in a maximum sample throughput frequency that is achieved at a single optimal thionine level and is independent of the analyte concentration. In contrast the optimal thionine concentration during continuous regeneration varies with the sample analyte concentration. These findings highlight the importance of designing controllable regeneration for dehydrogenase-based optical biosensors. PMID- 15899585 TI - Biological activity of interleukins-28 and -29: comparison with type I interferons. AB - Despite binding to receptors distinct from those of type I interferons (IFNs), human interleukins-28A, -28B and -29 (IL-28A, IL-28B and IL-29; alternatively named IFN lambda-2 {IFN-lambda2}, IFN-lambda3 and IFN-lambda1, respectively, or collectively, type III IFNs), a small family of three structurally-related cytokines, are, like IFNs, known to induce antiviral activity. To further biologically characterize IL-28A and IL-29, we compared their activities with those of IFNs in a range of human cell lines. We found that they induced antiviral activity in fewer cell lines and more weakly than IFNs; also IL-28A was less active than IL-29. Additionally, we showed IL-28A and IL-29 induced reporter genes--protein MxA promoter linked to luciferase, or interferon stimulated response element (ISRE) linked to secreted alkaline phosphatase (SEAP)--more weakly than IFN. Antiproliferative activity was induced by IFNs in most cell lines, but only in one human glioblastoma cell line, LN319, was dose-dependent IL 29-growth inhibition demonstrable. Polymerase chain reaction (PCR) quantification of messenger (m) RNA of IL-28/29 receptor subunits, IL-28Ralpha and IL-10Rbeta, indicated variable expression levels; although their expression was highest in the responsive LN319 cell line, lower but significant expression of both mRNAs was found in relatively unresponsive cell lines. In conclusion, we found IL-28A and IL-29 act similarly to IFNs, but are less effective generally and have activity in a more limited range of cell lines. PMID- 15899586 TI - The characterisation of six ADAMTS proteases in the basal chordate Ciona intestinalis provides new insights into the vertebrate ADAMTS family. AB - ADAMTS, constituting a recently discovered family of secreted zinc-dependent metalloproteases, have been shown to have critical physiological roles through identification of a number of natural animal and human gene mutations. The identification of six ADAMTS genes in the basal chordate Ciona intestinalis provides new insight into how, when and in what order the vertebrate orthologues have evolved. The phylogenetic assignments, based on sequences conserved across all genes, are supported by conserved domain structures within defined sub families. The phylogeny and the frequent localisation of ADAMTS genes in paralogous regions of the genome are consistent with the vertebrate lineages having arisen by large scale or genome duplication. The high level of conservation in the protease active site of vertebrate orthologues within some sub-families suggests subfunctionalisation, whereas the greater divergence in others would favour the evolution of novel substrate specificities and these observations are borne-out where substrate-specificity is known. The expansion and sub-specialization of the ADAMTS family is a component of the increased complexity of extracellular matrix that is associated with the evolution of vertebrates. PMID- 15899587 TI - Cell type-specific genotoxic effects of intermittent extremely low-frequency electromagnetic fields. AB - The issue of adverse health effects of extremely low-frequency electromagnetic fields (ELF-EMFs) is highly controversial. Contradictory results regarding the genotoxic potential of ELF-EMF have been reported in the literature. To test whether this controversy might reflect differences between the cellular targets examined we exposed cultured cells derived from different tissues to an intermittent ELF-EMF (50 Hz sinusoidal, 1 mT) for 1-24h. The alkaline and neutral comet assays were used to assess ELF-EMF-induced DNA strand breaks. We could identify three responder (human fibroblasts, human melanocytes, rat granulosa cells) and three non-responder cell types (human lymphocytes, human monocytes, human skeletal muscle cells), which points to the significance of the cell system used when investigating genotoxic effects of ELF-EMF. PMID- 15899588 TI - Evaluation of liver and peripheral blood micronucleus assays with 9 chemicals using young rats. A study by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study Group (MMS). AB - We conducted simultaneous liver and peripheral blood micronucleus assays in young rats with seven rodent hepatocarcinogens-4,4'-methylenedianiline (MDA), quinoline, o-toluidine, 4-chloro-o-phenylenediamine (CPDA), dimethylnitrosamine (DMN), p-dimethylaminoazobenzene (DAB), and di(2-ethylhexyl)phthalate (DEHP)-and two mutagenic chemicals-kojic acid and methylmethanesulfonate (MMS). Quinoline, DMN, and DAB were positive in the liver assay, while o-toluidine, kojic acid, DAB, and MMS were positive in the peripheral blood assay. o-Toluidine, kojic acid, and DAB are reportedly negative in mouse bone marrow micronucleus assays, indicating a species difference. Our results revealed a correlation between micronucleus induction in hepatocytes and hepatocarcinogenicity. This technique can be useful for the detection of micronucleus-inducing chemicals that require metabolic activation, and it enables simultaneous comparison of the micronucleus inducing potential of chemicals in the liver and peripheral blood in the same individual. PMID- 15899589 TI - Spectral studies, cyclic voltammetry and synthesis of cobalt(II) and ruthenium(III) complexes with symmetric and asymmetric ring containing membered N2S2, N4, and N5 donor macrocyclic ligands. AB - Reaction of divalent cobalt(II) and trivalent ruthenium(III) salts (NO3, SCN and SO4) with macrocyclic ligands L1, L2 and L3 having N2S2, N4 and N5 core, have been designed and carry out. All these three macrocyclic ligands and their complexes were obtained in pure form. Their structures were investigated by using microanalytical analyses, IR, mass, magnetic moments, electronic and EPR spectral studies. The redox properties of the complexes were also examined by cyclic voltammetry. An interesting feature of complexes is that the relatively large rings of macrocyclic ligands prevent the macrocyclic rings from approaching the metal center as closely as they would, if they were not constrained. So the Ru-N distances are longer than expected due to ring size. Electrochemical studies show that the macrocyclic ligand L1 is more effective electron donors to ruthenium than of L2 and L3. Electronic spectral properties also show that the sulphur donor atom of L1 weakens the ligand field with respect to ligand-to-metal charge transfer band. However it is expected that second-row transition metal-ligand bonds tend to be weaker than third-row transition metal-ligand bonds. There are well-established examples of reactions in which decreased of reactivity down a triad of transition metals is not observed. These novelties are usually attributed to pi-bonding effects for ligands such as carbon monoxide, solvent effects, or a change in mechanism. PMID- 15899590 TI - Formation of rat hepatocyte spheroids on agarose. AB - The hepatocyte spheroid formation on the Primariatrade mark dish (PM) occurred as a result of cell migration, adhesion, and shape changes. On the other hand, hepatocytes plated on 1.5% agarose-coated dishes did not attach to the dishes. The formation of spheroids on the agarose-coated dishes was significantly more rapid than that on the PM ones. The PA/plasmin system plays an important role in the process of hepatocyte spheroid formation on PM dishes, and thus the spheroid formation on PM is inhibited by plasmin inhibitors. Nevertheless, these inhibitors did not show any effect on the spheroid formation on agarose-coated dishes. PA activities could be detected in the conditioned medium harvested from the cells cultured on PM dishes, but not in that from those cultured on the agarose-coated dishes. In summary, we have established a novel method to form hepatocyte spheroids by using agarose-coated dishes, in which some mechanism(s) independent of the PA/plasmin system is involved in their formation. The differentiated hepatocyte-specific functions of the spheroids formed on the agarose-coated dishes were expressed to a quite lesser degree than those formed on the PM ones, suggesting that the processes involved in spheroid formation are important for the expression of spheroid functions. PMID- 15899591 TI - Delayed automatic detection of change in speech sounds in adults with autism: a magnetoencephalographic study. AB - OBJECTIVE: Autism is a form of pervasive developmental disorder in which dysfunction in interpersonal relationships and communication is fundamental. This study evaluated neurophysiological abnormalities at the basic level of language processing, i.e. automatic change detection of speech and non-speech sounds, using magnetoencephalographic recording of mismatch response elicited by change in vowels and tones. METHODS: The auditory magnetic mismatch field (MMF) was evaluated in 9 adults with autism and 19 control subjects using whole-head magnetoencephalography. The MMF in response to the duration change of a pure tone or vowel /a/ and that in response to across-phoneme change between vowels /a/ and /o/, were recorded. RESULTS: The groups were not significantly different in MMF power under any conditions. However, the autism group showed a left-biased latency prolongation of the MMF particularly under the across-phoneme change condition, and this latency delay was significantly associated with greater symptom severity. CONCLUSIONS: These results suggest that adults with autism are associated with delayed processing for automatic change detection of speech sounds. These electrophysiological abnormalities at the earliest level of information processing may contribute to the basis for language deficits observed in autism. SIGNIFICANCE: These results provide the first evidence for delayed latency of phonetic MMF in adults with autism. PMID- 15899592 TI - Inverse association between Slow Wave Activity per cycle and the number of ultradian sleep cycles per night in healthy humans. AB - OBJECTIVE: Comparisons of sleep Slow Wave Activity (SWA) during successive sleep cycles rely on the assumption that SWA in a given cycle is independent of the number of ultradian cycles present in a night. This assumption was evaluated here. METHODS: Twenty-six healthy controls with no medical, sleep or psychiatric disorders were selected among 84 candidates and their sleep was recorded at home across 2 consecutive nights after two habituation nights. RESULTS: In comparison with nights with less cycles, nights with more cycles showed significantly more REMS but not more NREMS. No correlation was found between the number of cycles and the integrated SWA per night (epochs visually scored as NREMS). However, inverse correlations were found between the number of cycles and the SWA per cycle. This was significant on both nights in Cycle 1 and strong trends were found for the two subsequent cycles on Night 2. Comparable results were found after removal of nights containing suspected Skipped First REMS episodes. CONCLUSIONS: The SWA in a cycle was found to be inversely correlated to the number of cycles in the first 3 cycles in at least one of the two analyzed nights. SIGNIFICANCE: Differences in the number of cycles per night are a potential bias in the comparisons of SWA per cycles. PMID- 15899593 TI - The use of complementary and alternative (non-conventional) medicine in cancer patients in Turkey. AB - The purpose of this study was to define various methods of complementary and alternative medicine (CAM) used by cancer patients and their characteristics in Turkey. The sample consisted of 100 cancer patients who were monitored in the oncology outpatient and inpatient departments of two hospitals. Almost all participating patients were receiving conventional treatment, 36% of which were also using CAM methods. Female patients and individuals with lower educational background showed a tendency to use CAM methods more often. The most frequently used CAM method appeared to be herbal therapy, and the most commonly used herb was the stinging nettle. Most patients obtained information about CAM methods from their friends, the media and from CAM vendors. Patients' responses indicate that "the desire to do everything possible to fight the disease" and "the idea that it may be helpful, at least it's not harmful" were the two most common reasons for using CAM. Of the patients, 79.3% acknowledged a varying degree of benefit from the methods they use. The most frequently indicated benefit was "feeling better both physically and emotionally". Most findings in this study are similar to the results of other research conducted on the subject in Turkey. PMID- 15899594 TI - Tissue Doppler imaging does not show infraclinical alteration of myocardial function after contrast echocardiography. AB - BACKGROUND: It has been previously suggested that simultaneous exposure of hearts to contrast and ultrasound can damage the myocardium and produce a transient decrease of the contractility in animals. Tissue Doppler imaging (TDI) is a useful tool to quantify the myocardial function with very high temporal resolution. AIM OF THE STUDY: The aim of the study was to test whether contrast echocardiography (CE) can cause alteration of the myocardial function by using tissue Doppler analysis. METHODS: Twenty-eight healthy patients (mean age: 44 +/- 22) underwent baseline echocardiography before and after 5 min of continuous intravenous infusion of Sonovue from the apical views, using an intermediate mechanical index (MI = 1). High frame rate images were acquired in tissue Doppler mode. Data were averaged over 3 cardiac cycles and analysed off-line before and after CE. RESULTS: There were no significant changes, before and after CE, in the peak systolic velocity (basal septum (BS): 6.2 +/- 2.2 vs 6.4 +/- 2.6; basal lateral (BL): 6.2 +/- 3.1 vs 6.4 +/- 3.3 cm/s), in the peak diastolic E velocity (BS: 5.4+/-1.8 vs 5.3+/-1.7; BL: 7.3+/-2.4 vs 7.7 +/- 3.2 cm/s), in the peak diastolic A velocity (BS: 6.3 +/- 1.9 vs 6.9 +/- 2.4; BL: 6.1 +/- 3.5 vs 6.2 +/- 2.5 cm/s), in the peak systolic strain (BS: 16 +/- 7 vs 17 +/- 7; BL: 12.6 +/- 5 vs 12.9 +/- 5%) and in peak systolic strain rate (BS: 1.3+/-0.6 vs 1.4+/-0.6; BL: 1.2+/-0.5 vs 1.21+/-0.51 1/sec). CONCLUSIONS: Our data suggest that CE does not cause alterations in the myocardial function as assessed by tissue Doppler imaging. CE, even with high MI settings, usually used for left ventricular opacification, can be safely performed. PMID- 15899595 TI - A thin layer chromatographic method for determining the enzymatic activity of peroxidases catalyzing the two-electron reduction of lipid hydroperoxides. AB - Thiol-dependent peroxidases catalyzing the reductive detoxification of lipid hydroperoxides (LOOHs) are crucial antioxidant components of mammalian cells. There is a growing interest in manipulating expression of such enzymes to better understand their biological roles. A new approach for determining their cellular activity is described, whereby LOOH reduction kinetics are tracked by high performance thin layer chromatography with peroxide-sensitive tetramethyl-p phenylenediamine detection (HPTLC-TPD). The approach was tested on a tumor cell transfectant clone (7G4) over-expressing selenoperoxidase GP x 4. Timed incubation of Triton-solubilized 7G4 cells with GSH and peroxidized phosphatidylcholine (PCOOH), followed by lipid extraction, HPTLC-TPD and densitometry revealed an exponential decay of PCOOH at a rate approximately 80 times greater than that for GP x 4-deficient controls (VC). A TPD-detectable cholesterol hydroperoxide (7alpha-OOH) was also reduced much faster by 7G4 than VC extracts. Spraying with H(2)SO(4) after TPD revealed both 7alpha-OOH loss and resolved diol product (7alpha-OH) accumulation, the kinetics of which were identical. The approach described is relatively convenient, highly specific, and much more sensitive than conventional assays for cellular LOOH reducing enzymes. PMID- 15899596 TI - Protein-inhibitor complexes analyzed by alkaline capillary LC-MS. AB - Liquid chromatography-mass spectrometry (LC-MS) has been used extensively in determination of the molecular weights of proteins, as well as covalent protein ligand complexes. We have successfully developed LC-MS method for protein molecular weight measurement using small-bore and capillary LC-MS under acidic and basic conditions. A high pH method was critical in studying complexes that were unstable under acidic conditions. Microgram sensitivity was achieved using both methods. A protocol to study the binding mode of protein-ligand complexes under denaturing conditions was developed. These methods were applied to CP88 (a proprietary cysteine protease) inhibitors and revealed different binding modes of inhibitors to proteins that had similar non-reversible behavior in biochemical activity assays. The method also confirmed that one inhibitor studied binds to CP88 in a reversible covalent manner. PMID- 15899597 TI - Determination of 3-methylhistidine and 1-methylhistidine in untreated urine samples by capillary electrophoresis. AB - Capillary electrophoretic (CE) method was developed for the determination of urinary 3-methylhistidine (3MH) and 1-methylhistidine (1MH) indicating the extent of degradation of skeletal muscle proteins and thereby the state of human health. 3MH, 1MH and histidine can be separated in both acidic and alkaline media, where these amino acids form cation and anion, respectively. The effective mobility of all ionic forms was measured over a broad range of pH (1.67-11.80), which made it possible to evaluate the corresponding dissociation constants. 3MH and 1MH were determined together with creatinine in untreated urine samples with the limit of detection of 2.4 microM (0.4 mgL(-1)) and 3.0 microM (0.5 mgL(-1)), respectively. Determination was fast and took ca. 12 min including the column washing. Method was employed for an analysis of urine collected from healthy individuals, and from the patients hospitalized with obesity and diabetes mellitus II. This analysis has revealed differences between the healthy individuals and the patients pointing to a more extensive degradation of muscle proteins in the latter group. PMID- 15899598 TI - Functional dyspepsia pathogenesis and therapeutic options--implications for management. AB - Functional dyspepsia is far more common than dyspepsia due to organic disease, both in the community and general practice. Proposed aetiopathogenic factors include gastric acid, Helicobacter pylori infection, delayed emptying, hypersensitivity or impaired accommodation of the stomach, dysfunction of the duodenum or brain-gut axis, psychosocial morbidity and post-infective mucosal damage. More effective therapy will depend on the development of drugs targeted at these putative pathophysiological mechanisms. On current evidence tricyclic antidepressants appear to be more effective than either acid suppressants or H. pylori eradication. PMID- 15899599 TI - [Alzheimer's disease]. PMID- 15899600 TI - [Psychobehavioral disorders in severe dementia]. AB - Behavioral and psychological symptoms of dementia (BPSD) are not included in the diagnostic criteria for dementia or in the assessment tools for grading its severity. However, they constitute a major part of the caregivers' burden and the main cause for institutionalization. They are, for one part, the direct consequence of the brain lesions, but, for another part, they result from the psychological reactions of the patients to their cognitive deficits or their relational difficulties with their family or carers. Despite speech disorder and mind disruption, patients with severe dementia still have a psychic and relational life, which is expressed through BPSD. Many BPSD are of short duration and can be improved by a better understanding, psychological and behavioral approach. However, psychotropic drugs are often useful or necessary, but, due to the frailty of the patients with severe dementia, their use should be very cautious. PMID- 15899601 TI - [Patients with severe dementia living at home]. AB - Few studies have been devoted to patients with severe dementia living at home. Therefore their rate is difficult to evaluate, and their conditions too heterogeneous to allow a standard management. In these patients, suffering from severe cognitive deficits and psychobehavioral disturbances, interfering pathologies may alter the course of the disease. Environment must be adapted to ensure security, sustain perceptual and physical activity and back up space and time orientation. The role of the main caregiver is crucial for the psychological and physical support of the patient. Thus, he/she must be sustained by giving information about the disease and available practical and financial support, and by listening to his/her suffering. Consequently, the general practitioner is involved not only in medical decisions but also in family support. Too often, the general practitioner is alone to face complex and time-consuming situations. However, the setting of geriatric networks should allow him/her to be included in the elaboration of a global care project with nurses and physiotherapists. Nevertheless, various factors, such as caregivers' exhaustion, major behavioral disorders, or the end of life when coping with pain becomes a priority, can eventually make necessary the patient's transfer into a geriatric facility. PMID- 15899602 TI - [Patients with severe dementia in geriatric facilities]. AB - The institutional management for demented patients has been improved by new specific treatments, behavioural approach, better architectural adaptation, and environmental characteristics. This management must be global, including patients, families and nursing staffs. The nursing staff must be diversified, motivated, and specifically trained to manage demented patients. PMID- 15899603 TI - [Nutrition and Alzheimer's disease]. AB - Weight loss is frequent in Alzheimer's disease. Its severity increases with the progression of the disease and may be a predictor of patients' mortality. Weight loss often precedes the diagnosis and may be considered as a feature of the disease itself. With the progression of the disease, disorders of eating behavior occur and result in weight loss and decreasing energy intake. Descriptive tools such as the Blandford Scale are helpful to assess eating disorders. Some hypotheses exist to explain the weight loss associated with Alzheimer's disease: atrophy of internal temporal cortex, increase of energy expenditure, biologic factors or modifications of body composition. Tools such as the Mini Nutritional Assessment allow assessment of nutritional status, diagnosis of malnutrition, cause identification and proposals for its correction. Patients' caregivers play a central role in prevention and management of nutritional disorders. Information programs for caregivers may reduce caregiver burden and improve patients' health status. PMID- 15899604 TI - [Non-pharmacologic approach in severe dementia]. AB - Care for patients with Alzheimer's disease, particularly with severe dementia, requires a global therapeutic strategy integrating pharmacological approach into the environmental dimensions, psychotherapeutics and rehabilitation. The objective is to maintain autonomy as long as possible but also to improve the quality of life by reducing the psychological suffering of patients and families. In severe dementia, behavioral techniques and organization of the environment are possible at home but, actually, most of the patients are institutionalized. Structures having a specific project of life, of care and specific architectural design should be preferred. Education and support for caregivers as well as training of the nursing staff are essential to develop better attitudes toward the patient, improve communication and optimize the quality of life. In spite of the low level of evidence in the evaluation of these strategies on account of the lack of adapted indicators and rarity of specific research, widely spread techniques can confer special purport to this difficult stage for the patients and their family. PMID- 15899605 TI - [Pharmacological treatment in severe dementia]. AB - Medical treatment of severe dementia is now available. Decrease of psychobehavioral disturbances, autonomy loss or care-giver burden appear as the main objective rather than reduction of cognitive deficits. Acetylcholinesterase inhibitors (AChE-I) should be maintained in patients with severe dementia when initiated in mild or moderate dementia. Memantine is the specific treatment for patients with severe dementia, even if they received AChE-I. To treat the psychobehavioral disturbancies, serotoninergic agents and thymoregulators are the first line drugs. Medical treatment should only be co-prescribed with a global care of all co-morbidities, autonomy loss and patient's and care-giver's burden, and associated with psychological and organisational support. PMID- 15899606 TI - [The cost of severe dementia]. AB - The aim of the present article is to analyse the methodological difficulties to assess the economic cost of severe dementia from the available data, and the shortcomings of an exclusively economic approach. It is very difficult to provide a reliable account of the cost of severe dementia, when ignoring the initial assessment of the severity and the socio-economic context of the care given to demented persons. When assessing the cost of severe dementia, one needs to know who are the affected individuals, and what medical care they are given - whether at home or inside an institution. While the direct cost can be easily assessed, the indirect cost is more difficult to calculate, and particularly the productivity loss for the caring persons, when they are in occupational activity. Setting up reliable indicators for the cost of dementia is a long process, which needs a clear definition of the target population of the measurement. PMID- 15899607 TI - [Ethical approach to severe dementia]. AB - General principles of medical ethics are discussed in the context of care for patients with severe dementia. PMID- 15899609 TI - [Classification of preinvasive breast and carcinoma in situ: doubts, controversies, and proposal for new categorizations]. AB - As a consequence of increased screening mammography, preinvasive breast lesions represent a growing percentage of breast pathology diagnoses. Intraductal epithelial proliferations of the breast are, at present, classified into three groups: usual epithelial hyperplasia, atypical ductal hyperplasia and ductal carcinoma in situ. The boundary between those two last entities is not clearly defined on a morphological point of view although the clinical management is different. Columnar atypical lesions or flat atypical hyperplasia, frequently observed near microcalcifications, are not homogeneously managed. Lobular lesions are classified under the category of lobular neoplasia which avoid interobserver variability. Molecular biology data and immunoprofiles isolate different lobular phenotypes and link them to invasive cancer. The different systems of classifications of those different entities, the most recent molecular biology datas, controversies and consensus are described in this manuscript. PMID- 15899610 TI - [Assessment of HER2 status in breast cancer]. AB - HER2-positive status is the sole criterion for identifying patients with breast cancer for Herceptin (trastuzumab) therapy. Accurate assessment of HER2 status is essential to ensure that all patients who may benefit from Herceptin are correctly identified. There are several assays available to determine HER2 status: the most common in routine clinical practice are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). The pros and cons of those two tests and the other upcoming methods for assessing HER2 status (with a focus on chromogenic in situ hybridization CISH recently approved by European Commission) are described in this manuscript. The importance of adhesion to quality assurance programs is underlined. Finally, the different national testing guidelines are discussed. PMID- 15899611 TI - [What is new about sentinel lymph node biopsy in breast cancer?]. AB - Sentinel lymph node biopsy concept was described in several reports since 1994. Technical and identification procedure, false negative risk, learning curve criteria and benefits concerning cancer stadification and postoperative morbidity were assessed. Peroperative and postoperative pathologic examination procedures are improving progressively but remains still under debate. Classification for micro-metastasis, isolated cells and macro-metastasis in sentinel lymph node was suggested but their immunohistochimic detection and prognostic impact are unclear. These parts are discussed in this article. PMID- 15899612 TI - [Sentinel lymph node negative breast cancer larger than 30 mm: is axillary lymph node dissection constantly necessary?]. AB - Benefits provided by sentinel lymph node biopsy (SLNB) include improvement of pathologic examination and lower rate of sequels. The aim of this study was to assess the accuracy of this procedure in large tumors. From march 1999 and december 2003, 663 patients were operated for a breast cancer and underwent a SLNB. All patients with tumor larger than 30 mm and/or with involved SLNB underwent a complete axillary dissection (AD). One hundred and sixteen patients (17.5%) with a tumor larger than 30 mm underwent an AD. Identification rate of SLNB was 94% (109/116). Among the 43 cases with non metastatic SLNB, the AD was negative in 39 cases (90.7%). In four cases a palpable non sentinel lymph node discovered during the SLNB procedure was found to be involved. In one case the lymphoscintigraphy found only one lymph node with a very low fixation and an AD was performed because of low efficiency of the lymphoscintigraphy procedure. SLNB false negative rate was 1.4 % (1/72) among the 72 cases with lymph node involvement. AD was performed in 66 cases (60.5%) during the same operating time than the SLNB. AD showed a lymph node involvement in 30 cases (30/72, 41.7%): in 5 cases among 24 cases with microscopic disease (20.8%) and in 25 cases among 48 cases with macroscopic disease (52%). This series results suggest than AD could be avoided for negative SLNB tumors up to 50 mm at pathologic examination in patients with clinically-measured tumor smaller than 30 mm. PMID- 15899613 TI - [Breast carcinoma and genomic technology: contribution of DNA microarrays]. AB - Genome wide expression analyses techniques are now applied to breast carcinoma. Several results have been achieved : molecular clustering of breast carcinomas, implementation of powerful prognostic classifications, and in some way predictive classification. Unfortunately, translation to bedside remains limited due to standardization difficulties. PMID- 15899614 TI - [Neoadjuvant treatment of early breast cancer: new drugs contribution]. AB - Neodjuvant chemotherapy has been primitively used in locally advanced breast cancer. More recently, indications have been extended to operable lesions. Several randomized trials have compared preoperative chemotherapy with postoperative surgery. None of these studies with anthracyclines have shown benefit in terms of progression free survival and/or overall survival. The rate of breast-conservative surgery seems slightly increased. However, in the NSABP 18 study, there was also a marginally significant increase in local recurrence when patients were converted from mastectomy lumpectomy to as originally planned. The introduction of taxanes in neoadjuvant chemotherapy protocol leads to a better clinical response and a complete histological responses increase when docetaxel is used. In only one study using docetaxel after anthracyclin treatment, a survival benefit is reported but it is a small study; a longer follow up and the updated results of the NSABP B27 trial are needed to confirm the impact of taxotere in the outcome of breast cancer. The integration of trastuzumab with taxane based chemotherapy has already shown high rate of histological complete responses. At least, neoadjuvant chemotherapy has the advantage to study biologic predictive factors of chemotherapy response. This article will review data on standard therapeutic and new agents in neoadjuvant treatment. PMID- 15899615 TI - [Aromatase inhibitors in adjuvant setting in breast cancer]. AB - For more than twenty years, tamoxifen represents the gold standard treatment in adjuvant setting for breast cancer patients. However, the tamoxifen activity remains complex, with its agonist effects, sometimes a poor tolerance and a certain number of patients become refractory to treatment. The aromatase inhibitors, such as progestatifs, were developed to challenge tamoxifen efficacy, along with improved tolerability. The third generation of aromatase inhibitors seems to provide significant gains in efficacy over tamoxifen for postmenopausal patients with hormone receptor positive breast cancer and they have already been approved in patients with metastatic disease. We review, in this article, the rationale for using IA in patients with breast cancer and, across the different clinical trials results already published, their current major role they are playing in adjuvant setting for menopausal hormonal receptor-positive breast cancer patient. One of the main issues using the third generation of IA is their long-term side effects, especially bone turnover and lipid metabolism. PMID- 15899616 TI - [Dose density and dose intensity in the treatment of breast cancer]. AB - Since 10 years, high-dose chemotherapy has been evaluated for the treatment of breast cancer in numerous randomized clinical trials. Preliminary results of some of these studies have shown an advantage in relapse-free survival in both metastatic and high-risk breast cancer. Although follow-up is short in most of the studies, no impact on overall survival has been detected. Based on available results, high-dose chemotherapy cannot be proposed either in metastatic or in high-risk breast cancer patients outside a clinical trial. Conversely, two randomized trials have demonstrated that dose-dense scheduled chemotherapy with G CSF support, containing an anthracycline, cyclophosphamide and paclitaxel, improves clinical outcomes compared with the same regimen administered every 3 weeks. These results establish dose-dense scheduled chemotherapy containing an anthracycline and paclitaxel as an option for the adjuvant treatment of positive lymph nodes breast cancer patients. Data are not sufficient to conclude in the neoadjuvant and metastatic setting. High-dose chemotherapy and dose-dense chemotherapy seem to increase the pathological complete response rate in inflammatory breast cancer. However, prospective and comparative survival data are lacking. PMID- 15899617 TI - [Expression of chemokine receptors by cancer cells]. AB - Chemokine receptors are involved in the trafficking of leucocytes. It has been shown recently that chemokine receptors (CXCR4, CCR7...) are expressed by breast cancer cells and could be involved in the metastasis phenomenon. Since this discovery, new data have been generated in this topic. These data confirm that chemokine receptors are involved in the occurrence of metastases, suggest that their expression is regulated by genes involved in the carcinogenesis and/or angiogenesis (VHL...) and that inhibitors could be of interest in the treatment of breast cancer. In this paper, we will review all these data and will discuss the perspectives that opens this new concept. PMID- 15899618 TI - [Targeted therapies: last focus on EGF receptor inhibitors in breast cancer]. AB - EGF receptor is slightly expressed in breast cancer without any sufficiently impact on patient outcome. Nevertheless determination of EGFR level represents a certain interest in order to orientate a targeted therapy notably in the case of hormone therapy resistance. Based on preclinical findings, preliminary clinical study results confirms in vitro observations about the possibility to encompass the hormonotherapy or anti-HER2 resistance. But, future large clinical trials are necessary to redefine a new strategy to treat breast cancer patients. PMID- 15899620 TI - [Malignant cardiac tumors]. AB - Cardiac neoplasms are a rare occurrence in clinical practice. The various frequencies of primary and secondary malignant tumors vary from report to report, approximately 1% in most autopsy series and 4% in cancer patient's autopsies. Cardiac malignancies account for less 1% of cardiac surgery and about for 0.1% of cardiac echographic studies. The presence of metastatic tumor to the heart usually indicates widespread metastases. Lung carcinomas are the most commonly encountered tumor followed by breast and pancreas cancer and melanoma. Apart from primary pericardial mesothelioma, primary cardiac tumors are high-grade sarcomas with a high metastatic potency that often becomes evident early after surgery. Symptoms are non specific, occur late in the disease and affect few patients; especially secondary neoplasms of the heart take their course so fast that they cannot become symptomatic. The signs of cardiac neoplasms are divided into systemic symptoms (fever, arthralgias and myalgias), cardiac symptoms (congestive heart failure, arrhythmia, chest pain) and uncommon embolisms. Diagnosis is actually made easier with cardiac echography. Cardiac RMI is helpful to estimate vessels and pericardium involvement. Due to its poor prognosis, treatment of cardiac metastases is restricted to best supportive care. For primary cardiac neoplasms, surgery must be carefully discussed because operative intervention is often followed by rapid widespread metastases that adjuvant chemotherapy cannot avoid in most cases. PMID- 15899621 TI - [Radiation toxicity to the heart: physiopathology and clinical data]. AB - Irradiation of the heart begets different complications in which pericardial disease is the most common manifestation. Thoracic irradiation of lymphoma, Hodgkin's disease, lung and breast cancer could be complicated by adverse effects to every structure of the heart. Potential injury of mediastinal irradiation can include acute and late pericarditis, cardiomyopathy, valvular disease and conduction abnormalities. The pathophysiology of these various syndromes is probably similar, starting by prior microvascular injury that leads to subsequent myocardium ischemia, all of which cause late fibrous scars. Acute pericarditis is often asymptomatic and clear spontaneously. Late pericarditis affects approximately 5% of the patients when the irradiation dose exceeds 40 Gy. At this dose, the mortality rate is below than 1%. Cardiomyopathy is rare and often asymptomatic. A long time unrecognized, coronary artery disease, diagnosed in 5 to 10% of the patients, begets multifarious sequelae like myocardial infarction, valvular abnormalities and cardiac rhythm changes. This coronary artery disease is more likely to occur if the patient was young at the time of the irradiation (< or = 21 years) and/or if other cardiovascular risk factors are associated. Incidence and mortality rates of valvular defects are about 20 and 0.5%, respectively. Conduction dysfunctions can also be seen in 5% of the patients. Radiation-induced heart complications seem to be related to total dose (> 30 Gy), irradiated tissue volume and fraction size. Since cardiac complications appear months to years following incidental irradiation of the heart, appropriate screening and long-term cardiac follow-up of these patients is essential. PMID- 15899622 TI - [Cardiotoxicity of 5-fluorouracil]. AB - 5-fluorouracil (5FU) is a largely employed antimetabolite, responsible for several well-known toxicities like hand-foot syndrome, diarrhoea, mucositis or leucopenia. Cardiotoxicity of 5-FU is known but uncommon and usually not life threatening. The incidence has varied from 1.5 and 18%. The physiopathology is controversial, although more recent data suggest a myocardial toxicity. Clinical presentation include chest-pain, cardiac arrythmia, myocardial infarction or global cardiac failure. Electrocardiographic features are usually aspecific, with ischemic signs. The main treatment is to stop the 5-FU infusion and to introduce symptomatic cardiologic treatment. Although the prognosis is good, the mortality ranges between 2.2 and 13% in case of symptomatic cardiotoxicity. Secondary prophylaxis remains controversial. PMID- 15899623 TI - [Anthracycline-induced cardiotoxicity]. AB - Anthracyclines are topoisomerase II inhibitors that are used for various malignancies, but their usefulness is limited by cardiotoxicity. Three types of cardiotoxicity are distinguished: acute, sub-acute, and chronic. The chronic toxicity is related to the peak plasma concentration and the cumulative dose. Generation of free radicals appears to be the main cause of myocardial damage in late-onset cardiotoxicity. Anthracycline toxicity is exacerbated in pediatric and elderly population. An increased toxicity is also observed after heart irradiation and with some chemotherapy combinations. Myocardial damages are asymptomatic for a long time, but echocardiography detects a reduction of left ventricular ejection fraction. Endomyocardial biopsy is the only specific test for early diagnosis of anthracycline-induced toxicity. This cardiotoxicity can be reduced by anthracycline administration modifications, pharmacological cardioprotection, and doxorubicin analogues with a better tolerance profile, as epirubicin or liposomal anthracyclines. PMID- 15899624 TI - [Cardiac dysfunction induced by trastuzumab]. AB - Trastuzumab combined to chemotherapy had demonstrated significant improvement of time to progression and survival in metastatic breast cancer patient overexpressing HER-2. Cardiac toxicity was an unexpected side effect. The incidence of cardiac dysfunction was dependent on prior or current anthracycline exposure. The mechanism of action involved for this cardiac toxicity is unclear. We discuss in this article the incidence, the physiopathology, the risk factors, the predictive factors and the patient management and on cardiotoxicity of trastuzumab. PMID- 15899625 TI - [Cardiovascular toxicity of some cancer agents (others than anthracyclines, fluoropyrimidines and trastuzumab)]. AB - Many others cancer agents can lead to cardiovascular toxicity apart from anthracyclines, 5-fluorouracil, trastuzumab and ionizing radiations. Paclitaxel, high-dose cyclophosphamide and ifosfamide, mitomycin and irinotecan cardiotoxicity is unusual but for certain. Hormonal therapy is extensively used in breast and prostate cancers. Thromboembolic and cardiovascular events are increased by estrogens, synthetic progestatives and tamoxifene. Because of major toxicities related to intravenous interleukin-2, subcutaneous interleukin-2 regimens have been developed, allowing an improvement of cardiovascular tolerability. Interferon is less frequently associated with cardiotoxicity. Before treatment initiation, risk factors favouring cardiac toxicity should be evaluated. These risk factors can be related to the cancer agent, to the patient or to the tumour. Uncommon toxic events in general population can be more frequent in high risk patients. Therapeutic option can be influenced by these uncommon toxic events in high risk patients. PMID- 15899626 TI - [Cardiac toxicity of cancer treatment regimes in children and adolescents: physiopathology, clinical data and the paediatric oncologist's point of view]. AB - Over the last 20 years, the increase in the cure rate of childhood cancer and leukemia of almost 80% has facilitated the observation of middle and long-term sequelae, particularly of cardiovascular origin; such after-effects are the consequence of cytotoxic damage to the cells of the cardiovascular system, in particular by anthracyclines and radiotherapy, and all the more so by their combined use. Such destructive lesions to myocytes greatly hinder the capacity of the cardiac muscle to hypertrophy to meet the needs of bodily growth, pregnancy and certain intense sports activities. Endothelial cells also accelerate an early arteriosclerotic process, a potential cause of sudden death in the case of ostial stenosis. All such phenomena build up over time, together with the usual adult cardio-vascular risk factors. Finally, no cardiac tissue, pericardial, valvular endocardium or autonomic nervous tissue escapes these cytotoxic effects, giving rise to pericarditis, calcification, valvular leaks and arrythmias and conduction abnormalities. The resulting excessive cardiac mortality is one of the major concerns of paediatric oncologists, along with secondary tumours and leukaemia. This article analyses physiopathological consequences that are often asymptomatic or clinical, together with diagnostic, screening and follow-up methods for these patients, encouraging lifestyle modifications where appropriate or, when possible, treatment before the appearance of cardiac failure, myocardial infarction or sudden death. Other cytotoxic drugs such as high-dose cyclophosphamide, amsacrin, 5-FU and tubulin acting agents are also mentioned as a result of their cardiac toxicity, but this is not usually dose-cumulative. PMID- 15899627 TI - [Cardiac prevention: the oncologic point of view]. AB - Between the different anticancer treatments, radiation therapy, chemotherapies and some target therapies could have cardiac toxicity. This toxicity could be a clinical cardiac insufficiency reducing the global benefit of these treatments. Between the different anticancer drugs the more cardiotoxic and frequently used are anthracyclines. The combination of anthracyclines with other agents like trastuzumab or paclitaxel could improve the treatment results, but with an increasing risk of cardiac toxicity. Different strategies have been developed and evaluated concerning the prevention of cardiac toxicity induced by anthracyclines. They are: using of potentially less cardiotoxic anthracyclines but with no direct comparison in adjuvant trials; modification of anthracycline infusion not so used because of technical problems; developing therapeutic strategies who could offer the opportunity to optimize the combination of different treatment with no increasing of cardiac toxicity. Use of a cardioprotecting agent before anthracycline infusion (as dexrazoxane is) is possible. However, the indication is not well defined despite existence of efficacy results. PMID- 15899628 TI - [Cyclo-oxygenase 2 inhibitors and cancer]. PMID- 15899629 TI - [Cyclooxygenase 2 and carcinogenesis]. AB - The membrane glycoprotein Cox2 is regulated at transcriptional and post transcriptional levels by pro-inflammatory agents, cytokines, growth factors, oncogenes, and tumor-promoters. Cox2 is expressed during early stages of colorectal carcinogenesis from the premalignant adenoma stage, and adenocarcinomas of stomach, colon, breast, lung and prostate. Its expression is detected in neoplastic, inflammatory, endothelial and stromal cells. Cox2 is involved in the conversion of arachidonic acid into prostaglandins and thromboxanes, as well as the synthesis of malonaldehyde (MDA, a mutagen) and the production of hydrogen peroxide, which promotes carcinogenesis. The Cox2 products act in turn on serpentine receptors coupled to heterotrimeric G-proteins (R-TXA2, R-PG) that are connected to signaling elements implicated in oncogenesis. Thus, Cox2 plays a key role in early stages of carcinogenesis by promoting the proliferation of tumoral cells and their resistance to apoptosis, as well as angiogenesis, tumor cell invasion and setting up of the metastatic process. These mechanisms establish the rationale behind the therapeutic targeting of Cox2 in human solid tumors. PMID- 15899630 TI - [Cyclooxygenase 2 inhibitors and cancer chemoprevention]. AB - Chemoprevention is the use of natural or synthetic compounds in order to reverse, suppress or prevent the carcinogenic process. Among the many pathways dysregulated during the carcinogenic process, cyclooxygenase-2 (Cox2) seems to be one of the most promising pathways to target in order to achieve chemopreventive and anticancer effects. Indeed, Cox2 overexpression contributes to the carcinogenic by at least 5 different mechanisms including transformation of procarcinogens on carcinogens, pro-inflammatory and immunomodulatory effect, resistance to apoptosis, angiogenesis and invasion progression... This review will focus on the rationale and the ongoing research areas related to chemopreventive approaches targeting Cox2. PMID- 15899631 TI - [Cyclooxygenase 2 inhibitors and adenomatous polyposis coli]. AB - Prolonged use of aspirin and?or non-steroidal anti-inflammatory drugs induces a partial regression of either sporadic adenomas or adenomas in adenomatous polyposis coli (APC), but also their emergence and colonic cancer development in sporadic adenomas. Specific inhibitors of cyclooxygenase of type 2 (Cox2) induce less upper and lower digestive tract adverse events that non-specific anti inflammatory drugs. This better tolerance might allow a long-lasting use in patients with APC. At time, we don't know if such treatments are able to prevent the development of cancer in the rectum or duodenum of these patients. In this paper we will discuss the scientific proofs and potential interest of Cox2 inhibitors in the treatment of PAF. PMID- 15899632 TI - [Cyclooxygenase 2 inhibitors and colorectal cancer]. AB - Cyclooxygenase-2 (Cox2) is an inductible isoenzyme of cyclooxygenase undetectable in normal colonic mucosa and overexpressed in 80% colonic tumor. Several works in vitro and in vivo showed that Cox2 plays a key role in the multistep process of colorectal tumorigenesis such apoptosis inhibition of cellular proliferation and angiogenesis induction. So that Cox2 represent a potential molecular target in colorectal management and specific Cox2 inhibitors may be useful as chemopreventive as well as therapeutic agent in humans. In animals study Cox2 inhibitors was shown to be effective and in humans Cox2 inhibitors are approved by the Food and Drug Administration as an adjunct to endoscopic surveillance and surgery in patients with Familial Adenomatous Polyposis (FAP). The purpose of this article is to review the relationship between Cox2?Cox2 inhibitors and differents signaling pathways of colorectal carcinogenesis and to precise their possible molecular mechanisms of action. This work although review clinicals data of their efficacy as chemopreventive agent as well as therapeutic in the differents group at risk for colorectal cancer. PMID- 15899633 TI - [Cyclooxygenase 2 and breast cancer. From biological concepts to clinical trials]. AB - Cyclooxygenases (Cox) are prostaglandin synthetase enzymes which play a key role in mammary carcinogenesis. Several connections were demonstrated between Cox and a few oncogenes (v-src, v-Ha-ras, HER-2?neu, Wnt, p53 mutated), alimentary products (PUFAs), transcription factors (c-jun and c-fos), proapoptotic proteins [Bax et Bcl-x(L)] or antiapoptotic (Bcl-2), CYP19 aromatase gene, NFkappaB receptor (RANKL), angiogenesis (via VEGF, TXA2, oxid nitric synthetase, alphaVbeta3 integrin receptor), peroxisome gamma proliferator receptor (PPARgamma) and its ligand PGJ2 and with antitubuline chemotherapy drugs. No correlation of Cox2 expression with hormonal receptors was shown. In epidemiologic studies there is evidence of breast cancer risk reduction for women who take AINS for a lon time. Alimentary factors like resveratrol or insaturated fat acid reduce Cox2 expression in animal and could be investigated in human studies. Clinical trials are planed with the anti Cox2 celecoxib for breast cancer prevention, in adjuvant setting, in metastatic situation combined with exemestane or antitubulin drugs or in neoadjuvant therapy. PMID- 15899634 TI - [Cyclooxygenase 2 inhibitors and lung carcinoma]. AB - Cyclooxygenase-2 (Cox2) may play a crucial role in lung carcinogenesis. In fact, overexpression of Cox2 is common in non-small cell lung cancer (NSCLC) and seems to be associated with tumor progression, invasion and metastasis. In experimental animal models Cox2 has been shown to be involved in tumor angiogenesis, suggesting that Cox2 is a potential target for NSCLC therapy. Several in vivo studies have already shown that Cox2 specific inhibitors (celecoxib, rofecoxib) have anti-tumor activity and clinical trials using Cox2 inhibitors are currently ongoing in patients with NSCLC. PMID- 15899635 TI - [Cyclooxygenase 2 inhibitors and urologic and gynaecologic cancers]. AB - PGE2 is one of the most important prostaglandin involved in the oncogenesis. PGE2 is found at high concentration level in the most of epithelial cancer. Urologic and gynaecologic cancer express the enzyme which are at the origin of PGE2: cyclooxygenase 2. Cox2 inhibitors present anticancer properties demonstrated in wide varieties of cellular and animal models. Human applications are currently tested in many clinical trials for bladder, prostate and uterine carcinomas. PMID- 15899636 TI - [Safety of selective cyclooxygenase-2 inhibitors taken for a long time]. AB - The serious digestive side effects of the selective inhibitors the inducible cyclooxygenase-2 are reduced by 60% as compared to the nonselective non-steroidal anti-inflammatory drugs. The main risk factors associated with gastro-intestinal ulcers caused by the latter were found also with the selective inhibitors taken for long period (age > 60 years, antecedents of gastro-duodenal ulcers, concomitant aspirin treatment). In contrast, H. pylori infection was not found as risk factor apart from past history of gastro-duodenal ulcers. The complications in the lower digestive tract are twice less frequent with the selective inhibitors than with nonselective anti-inflammatory drugs. Nevertheless, it seems that the risk of exacerbation of inflammatory colitis is not reduced. The cardiovascular complications are discussed. Rofecoxib taken at supra-therapeutic dosage was recognised to increase the incidence of myocardial infarction. A such increase was not found with usual dosage or with celecoxib. The selective inhibitors may reduce the renal sodium excretion and increase the blood pressure, particularly in hypertensive patients whose the blood pressure has to be regularly checked. PMID- 15899637 TI - [Analgesic effects of cyclooxygenase 2 inhibitors]. AB - Traditional pain management strategies for cancer pain have relied on the use of opioids, nonsteroidal anti-inflammatory drugs (NSAIDs) and other adjuvant analgesics. However, the substantial adverse effects associated with their use has left many patients without dependable options for effective treatment. Recent advances in the understanding of pain and its pathophysiologic mechanisms have led to the development of novel therapeutics. Cyclooxygenase(Cox)-2-specific inhibitors (coxib) have an established efficacy in the treatment of chronic and acute pain comparable to that of traditional NSAIDs without the degree of gastrointestinal complication or the platelets inhibitor effect of traditional NSAIDs. Numerous studies have shown that coxibs are efficacious in the management of chronic and acute pain in various clinical settings including postoperative pain. The superior safety profile of coxibs in conjunction with a comparable efficacy to nonselective NSAIDs supports the use of coxibs in balanced analgesic regimens and provides the potential to incorporate coxibs into the pain management algorithm used to treat cancer pain. PMID- 15899638 TI - [Depression in the aged patient]. PMID- 15899639 TI - [Descriptive epidemiology and risk factors of depression in the elderly]. AB - Depressive symptoms are frequent among the elderly, and seriously impair their quality of life. The prevalence of depressive symptoms is difficult to evaluate on account of several epidemiological problems, such as the various definitions of "depression", which often prevent comparisons between studies. The second major difficulty is related to the selection bias due to differences in populations studied, and to the high risk of refusal to participate among the depressed subjects. Nevertheless, the prevalence has been estimated, in general population, from 1 to 4% for major depression according to the DSM-IV criteria, and from 8 to 16% for depressive symptoms which may require treatment. The heterogeneity of the symptomatology of depression in the elderly, with atypical clinical feature, leads to underestimate its diagnosis and lack of proper medical care. However, effective treatments exist and most cases of depression among elderly are treatable. In the literature, many risk factors of depression among the elderly have been identified : female sex, low socio-economical level, bereavement, prior depression, co-morbidities, disability, cognitive deterioration and vascular factors. To decrease the consequences of depression and to improve the quality of life of the elderly, the screening and treatment of depression should be strengthened in the general elderly population. PMID- 15899640 TI - [Evaluation of the depressive symptomatology in the elderly]. AB - Depressive symptoms are frequent in old age. They are associated with several types of mental disorders : major depression, bipolar disorder, dysthymia, organic mood disorder, mixed anxious and depressive disorder. Various types of assessment tools can help to evaluate depressive symptomatology in the elderly: self administered questionnaires, observer ratings, standardised interviews. All are helpful in the clinical approach of depressive states. Self-administered questionnaires are the best tools for a systematic brief screening for depression, while observer ratings will be used for a quantification of depressive symptomatology in the follow up or to evaluate the efficacy of therapeutic interventions. Lastly, standardised interview are mostly used in research protocols, taking advantage of their high specificity. For the assessment of depression as one of the behavioural and psychological symptoms of dementia, multidimensional tools like the NPI, or unidimensional instruments like the Cornell scale for depression will be preferred. PMID- 15899641 TI - [Clinical aspects of the depression in the elderly]. AB - Depression is the most frequent psychiatric disorder in the aged and a major cause of dependence. Its prognosis is poor on account of its consequences on physical health, the risk of suicide and of chronic evolution. It severely impairs the quality of life of the patients and makes a major contribution to the cost of public health since depressed subjects are hospitalized and use medical care more frequently than non depressed patients. However, they do not receive proper treatment for their depression. There is a large agreement on the necessity to improve the diagnosis of depression in the aged and to provide its adequate management. The difficulty of its diagnosis is not related to some specificity of depression but to the attitude of physicians toward aging. The recent guidelines for the diagnosis of depression insist on the indepency of the criteria regarding to age. The specificity of depression in the aged is only related to some clinical aspects and factors related to aging which interfere with its recognition, especially significant somatic complaints, cognitive disturbances and anxiety. Depression is no part of normal aging, but many risk factors are associated with aging, especially bereavement. A close clinical approach is required to distinguish depression from bereavement. PMID- 15899642 TI - [Suicide and depression in the elderly]. AB - Twenty per cent of the annual 12,000 deaths by suicide in France occur in subjects aged over 65. The rate of suicide by age in the population increases with aging. Aged people usually commit suicide by more violent means than younger people. Overall, the risk of suicide is multiplied by 20 in depressed subjects and by 40 in case of previous suicide attempt. This risk is probably higher in the aged. Suicide attempts are always to be sternly considered in aged people on account of the risk of a second successful attempt but also of a diminished life expectancy due to somatic as well as psychiatric reasons. Suicidal ideas are frequent in aged subjects but they strongly suggest depression which should be actively investigated. The so-called existential suicide, expression of the free will of subjects devoid of any psychiatric disorder who want, willingly and consciously, make an end to their life, probably exists. However, the high frequency of depression associated with suicide induces to be especially watchful in case of suicide ideas or previous suicide attempt in aged subjects. PMID- 15899643 TI - [Depression and dementia]. AB - Data from the literature devoted to the relationships between dementia and depression are controversial on account of numerous methodological biases (community studies or from neurological or psychiatric departments), categorical versus dimensional approaches and variability of assessment tools for depression, aim of the study (depression versus dementia or versus Alzheimer's disease, AD). The difficulty to discriminate depression from AD is largely overestimated due to the confusion between depression, depressive symptomatology and apathy. The distinction is greatly facilitated by taking into account the qualitative differences of the memory deficits and cerebral imagery. Distinction of depression from frontotemporal or subcortical dementias could be much more difficult. Relationships between depression and AD are controversial. Most reports of depression as a risk factor for AD in the subsequent years, actually describe depressed symptomatology linked to apathy in preclinical AD. However, some studies found a relationship between AD and depression occurring more than 10 years before the onset of AD symptomatology, suggesting some common risk factors. The so-called symptoms of depression in AD are more related to apathy and affective disturbances than to dysphoria. The frequency of major depressive episode (MDE), greatly varies according to studies, but the frequency of suicide is low. Depression in dementia is related to neurobiological factors as well as to psychological mechanisms. Therefore, its treatment should associate antidepressant drugs and psychological support directed to the patient and family. PMID- 15899644 TI - [Depression and organic brain diseases in the elderly]. AB - Depressive symptoms are observed in many organic brain diseases in the elderly, particularly in stroke, degenerative or vascular dementias and Parkinson's disease. In many cases, an accurate estimation of the respective part of neurobiological abnormalities, adjustment disorders, disability and narcissistic wounds related to the disease appears very difficult for the practitioner. Specific data on the therapeutical aspects of secondary depressive disorders remains quite scarce. The efficacy of antidepressant drugs may be less important in geriatric depression with cerebral disorders or "secondary depressions" than in primary ones. Consequently, electroconvulsive-therapy may appear as an interesting therapeutical option for these patients. PMID- 15899645 TI - [Prognosis of depression in the elderly]. AB - The prognosis of depression, when the depression is diagnosed, is rather similar in the elderly and in younger adults. Recovery, after the initial episode or after one relapse or recurrence, is observed in about two thirds of the patients. However, depression increases the vulnerability of the elderly and is associated with an elevation of mortality, mainly due to the occurrence of somatic pathologies, which could be facilitated by the depression. Relapses, recurrences, chronicity and absence of therapeutic response are associated with inadequate treatment or follow-up. The prognosis of depression in elderly people requires a systematic diagnostic research and an appropriate treatment. The importance of follow-up and of a psychotherapeutic approach associated with antidepressant drugs should be emphasized. PMID- 15899646 TI - [Biological treatment of depression in the elderly]. AB - Depression has a high prevalence and a poor outcome in the aged. However, it is often under diagnosed and under treated although treatment with antidepressants at optimal dose and duration can be effective on full remission. Numerous antidepressants are effective but most of them produce side effects. The choice of antidepressants must take into account the effect of age on side-effects and pharmacokinetics. IRSS or moclobemide are recommended as first-choice antidepressants because of a presumed side effect better profile. A dimensional approach based on neurobiological hypothesis of depression, specific clinical features of late life depression (e.g. psychomotor retardation) and biological target of the antidepressants should promote treatment efficacy. PMID- 15899647 TI - [Depression in the elderly; future prospects]. AB - Extensive research has been achieved on the old age depression during the last decades. Specific clinical features and management have been reported. Indirect clinical and behavioural indicators, and even biological ones should be considered for the detection of depression. Cognitive assessment must be promoted in standard practice to differentiate depression from incipient dementia, but also in depression occurring in long stay care, vascular depression and in the depression-executive dysfunction syndrome. Distinction between recurrent depressive episode and late-onset depression has to be clarified. Therapeutic strategies have to be reconsidered in duration, according to the trend toward chronicity of late life depression. They remain based on the use of pluri aminergic antidepressants. Depressive disorder in the elderly is associated with hippocampus dysfunction, but other biological variables should be taken into account according to a dynamic stress-vulnerability model. PMID- 15899648 TI - Autoantibodies against intrinsic factor (IF) measured with an ELISA using recombinant human IF as both catching and detecting reagent. AB - The aim was to develop and validate an assay for measuring autoantibodies against human intrinsic factor (IF). For this purpose 1.25 pmol of recombinant IF was coupled to each well. Samples (100 microL of plasma diluted 1:1) were incubated for 1 h, followed by 30 min with the detection reagent (biotinylated IF, 3.3 pmol/mL). Samples were obtained from healthy blood donors (n = 141) and patients with suspected vitamin B(12) deficiency (n = 355). The initial assay results (n = 99 samples) were in reasonable agreement with those obtained using a commercial assay (Diagnostic Products Corporation). All donors but one showed a negative result. For the patient populations the fraction of positive results decreased with increasing levels of serum vitamin B 12 : 0.67 for <100 pmol/L (n = 13), 0.17 for 100-150 pmol/L (n = 23), 0.06 for 151-200 pmol/L (n = 65) and <0.01 for >200 pmol/L (n = 254). The imprecision was 11% as judged by repeat analyses. All samples remained positive when the catching reagent was exchanged with vitamin B 12 -saturated IF, suggesting that the samples contained both blocking (type I) and non-blocking (type II)antibodies. Results obtained with IF as the detection reagent correlated to those obtained when anti-immunoglobulin G was used instead. In conclusion, an ELISA using recombinant IF as both catching and detection reagent seems suitable for the detection of IF autoantibodies in plasma. PMID- 15899649 TI - Antibody titer against malondialdehyde-modified LDL compares with HDL cholesterol concentration in identifying angiographically verified coronary artery disease. Comparison of tests by ROC analysis. AB - Antibody titer against malondialdehyde (MDA)-modified low-density lipoprotein (LDL) has been found to be associated with atherosclerosis, but it has not been established whether it would detect subjects with coronary artery disease (CAD). In the present study, receiver-operating characteristic (ROC) analysis was used to compare the diagnostic accuracy of the antibody titer against MDA-modified LDL and high-density lipoprotein (HDL) and LDL cholesterol levels in discrimination between subjects with (n = 51) and without (n = 35) angiographically verified 3 vessel CAD. As a result, the antibody titer against MDA-modified LDL was lower in subjects with CAD compared with subjects without CAD (p < 0.0001). The area under the ROC plot was 0.822 (95% CI, 0.727 to 0.918) for the antibody titer and 0.769 (95% CI, 0.661 to 0.876) for the HDL cholesterol concentration. Both the antibody titer and the plasma HDL cholesterol level were more accurate markers of CAD than the LDL cholesterol level. As a conclusion, our results indicate that the antibody titer against MDA-modified LDL discriminates between subjects with widespread CAD and those without CAD similarly as the HDL cholesterol concentration. Moreover, the antibody titer against MDA-modified LDL is inversely correlated with the risk of severe CAD. PMID- 15899650 TI - Fucosylation of serum glycoproteins in lung cancer patients. AB - Increased expression of sialyl Lewis X or A antigens on metastatic cancer cells leads to their selectin-mediated extravasation. Profound fucosylation of the serum microenvironment may be a factor that interrupts adhesion and influences the formation of metastases. In this study we quantitatively analyzed fucosylation of serum glycoproteins in small-cell and non-small-cell lung cancer patients. Fucosylation of four chosen glycoprotein bands was measured as the reactivity with Aleuria aurantia lectin on nitrocellulose blots, preceded by polyacrylamide gel electrophoresis. Relative fucosylation and fucosylation coefficients were calculated by densitometric analysis. Fucosylated oligosaccharides were observed in higher amounts in cancer sera when compared to sera from healthy individuals in all bands analyzed. Glycoproteins of a molecular mass of 29 kDa appear to carry more fucose residues than the 42-kDa band, comprising alpha(1)-acid glycoprotein and haptoglobin. Glycans of the 26-kDa band were fucosylated to a higher extent in non-small-cell vs. small-cell lung cancer. The results suggest that the extent of fucosylation could be a useful marker for estimation of the glycosylation status of serum proteins in cancer patients. Cluster analysis leads to the preliminary suggestion that the fucosylation status could serve as a predictive factor for patient survival. PMID- 15899651 TI - Association of cytochrome P450 2E1 genetic polymorphisms with squamous cell carcinoma of the oesophagus. AB - Squamous cell carcinoma of the oesophagus is one of the most common cancers among black males in South Africa. Genetic polymorphism in the cytochrome P450 2E1 (CYP2E1) gene, coding for one of the main enzymes involved in the bioactivation of tobacco- and alcohol-related substances, was investigated for its role in the development of oesophageal cancer. Three single nucleotide polymorphisms -1053C- > T, -1293G--> A (both give rise to CYP2E1*5 ) and 7632T--> A ( CYP2E1*6 ) in the 5'-untranslated region of CYP2E1 were investigated in 189 patients and 198 control individuals in South Africa. The mutant variants occurred between patients and controls at frequencies of 1% and 2% (-1053C--> T), 1% and 3% ( 1293G--> A) and 18% and 7% (7632T--> A), respectively. In comparing patients with controls, the heterozygous CYP2E1*6 genotype was associated with increased risk of the development of squamous cell carcinoma of the oesophagus (odds ratio, 5.90; p < 0.001) after adjusting for age, sex, smoking and alcohol consumption. In contrast, we did not find a significant association between CYP2E1*5 and oesophageal cancer. Six novel mutations, -1371G--> A, -1359C--> G, -1342C--> G, 1261T--> del, -1189T--> C and -1103C--> G, were identified by DNA sequence analysis of the CYP2E1 promoter region. In summary, our case-control study of oesophageal cancer revealed an elevated risk associated with the CYP2E1*6 allele in South Africans. PMID- 15899652 TI - alpha-Tocopherol as an antiretroviral therapy supplement for HIV-1-infected patients for increased lymphocyte viability. AB - The aim of our study was to evaluate the benefits of supplementation with 800 mg/day of alpha-tocopherol with regard to cellular viability in HIV-1 seropositive patients undergoing anti-retroviral therapy. A total of 29 patients participated in the study, of whom 14 were given the supplement and 15 a placebo. The analyses were carried out before treatment commenced and after 60, 120 and 180 days. The plasma levels of HIV-1 RNA showed a significant decrease as a consequence of treatment time in the groups studied (p = 0.0001), although the difference between the treatments over time was not verified (p = 0.7343). The percentage of viable lymphocytes showed a significant increase as a consequence of treatment time in both groups studied (p = 0.0002) and a significant difference between the treatments over time (p = 0.0472). The percentage of lymphocytes in apoptosis showed a significant reduction over time (p = 0.0003), as well as a significant difference between the treatments over time (p = 0.0321). The significant increase in cellular viability indicates that supplementation with alpha-tocopherol offers an additional positive effect on cellular preservation in HIV-1 individuals undergoing anti-retroviral therapy; however, it represents an additional risk of anti-retroviral therapeutic failure, possibly due to drug-drug interaction involving up-regulation of metabolic clearance. PMID- 15899653 TI - The effect of antioxidant supplementation on superoxide dismutase activity, Cu and Zn levels, and total antioxidant status in erythrocytes of patients with Graves' disease. AB - The effects of supplementation with a fixed combination of antioxidants (vitamins C and E, beta-carotene and selenium) on superoxide dismutase activity, copper and zinc concentrations, and total antioxidant status were monitored in erythrocytes derived from a group of patients with Graves' disease treated with methimazole, with respect to the rate of achieving euthyroidism. Thyroid-stimulating hormone (TSH), thyroid hormones and the above-mentioned parameters were measured before therapy, and on days 30 and 60 after therapy initiation. The patients receiving antioxidant supplementation along with methimazole therapy (group A, n = 27) achieved euthyroidism at a faster rate than those treated with methimazole alone (group B, n = 28). The activity of superoxide dismutase decreased significantly in both patient groups during the treatment; however, there was no significant difference between the groups. There was no significant change in the erythrocyte concentration of copper, whereas the zinc concentration and total antioxidant status showed significant between-group differences. The study results clearly show that antioxidant supplementation in the treatment of Graves' disease is justified, while zinc and total antioxidant status in erythrocytes seem to be sensitive indicators of the efficacy of supplemental therapy. PMID- 15899654 TI - Concentrations of calcium, copper, iron, magnesium, potassium, sodium and zinc in adult female hair with different body mass indexes in Taiwan. AB - We investigated concentrations of calcium, copper, iron, magnesium, potassium, sodium and zinc using atomic absorption spectroscopy in the hair of four groups of adult females (n = 392), ranging in age from 20 to 50 years, with different body mass index (BMI): BMI < 18, slim group; BMI 18-25, normal group; BMI 26-35, overweight or obese group; and BMI>35, morbidly obese group. We found that the group with BMI < 18 had the highest ratios for [Ca]/[Mg], [Fe]/[Cu] and [Zn]/[Cu], but the lowest ratio for [K]/[Na] in hair. On the contrary, the group with BMI > 35 had the highest ratio for [K]/[Na], but the lowest for [Fe]/[Cu] and [Zn]/[Cu] in hair. Furthermore, when we compared concentrations of Ca, Cu, Fe, Mg, K, Na and Zn between the groups with BMI < 18 and BMI > 35, we found that there were significant differences (p < 0.05) in zinc concentrations between these two groups. In addition, there were significant differences in Ca, Cu, Mg, K and Na concentrations, with p < 0.01 at least. From this point of view, we suggest that hair concentrations of Ca, Cu, Fe, Mg, K, Na and Zn may be correlated with adult female BMI, but further studies are needed. PMID- 15899655 TI - Estimation of the limit of detection with a bootstrap-derived standard error by a partly non-parametric approach. Application to HPLC drug assays. AB - A recently proposed procedure for estimating the limit of detection (LoD) of an assay using a partly non-parametric approach is applied to five HPLC drug assays. The non-parametrically determined 95th percentile of the blank measurements (LoB) is obtained as the value of the N (95/100) + 0.5 ordered observations. The LoD is the lowest level of analyte that is likely to yield a measured result exceeding the LoB. The LoD equals LoB + c(beta) x SD(S), where SD S is the analytical SD at low concentrations, and c(beta) = z(1-beta)/(1-1/(4 x f )) (f = degrees of freedom). c(beta) approximately 1.65 for a type II error of 5%. The blank distributions deviated significantly from normality for four of the five assays because of skewness to the right. The estimated LoB ranged from 3.3 to 10.2 nmol/L. The LoDs were in the interval from 7.8 to 17.2 nmol/L. A new procedure for estimation of the standard error (SE) of the estimated LoD that is partly based on the bootstrap principle showed good performance in simulation studies. In conclusion, the partly non-parametric procedure for estimation of the LoD of typical HPLC drug assays was found to be a suitable approach. PMID- 15899656 TI - Pitfall in the high-throughput quantification of whole blood cyclosporin A using liquid chromatography-tandem mass spectrometry. AB - In a growing number of laboratories the technique of liquid chromatography-tandem mass spectrometry is used for the quantification of cyclosporin A in whole blood, employing cyclosporin D as the internal standard. Cyclosporin A is extensively metabolized in vivo; in liquid chromatography-tandem mass spectrometry respective metabolites can give rise to both parent and product ions that are isobaric with ions commonly used for the detection of cyclosporin A and cyclosporin D, respectively. In this article it is demonstrated that limited chromatography with co-elution of such metabolites together with cyclosporin A and cyclosporin D can lead to incorrect results. PMID- 15899657 TI - How accurate are clinical activity indices for scoring of disease activity in inflammatory bowel disease (IBD)? AB - Clinical activity indices are essential instruments in monitoring inflammatory bowel diseases such as Crohn's disease (CD) and ulcerative colitis (UC). To subclassify components of disease indices in CD and UC, investigate technical noise in estimation of the indices, establish a signal-to-noise ratio (SNR), evaluate correlation between indices and calculate the reference change value (RCV) for selected biochemical variables in individual cases, 50 patients with CD and 49 patients with UC were included in the study. Qualitative index variables were assessed for scoring errors. The standard deviation (SD) was estimated according to a rectangular model, while SD in biochemical variable scoring was estimated according to a Gaussian model; a combined SD was also calculated. These values were investigated for their individual contribution to variation. The 95% CI of an index value was based on +/- 1.96 x SD(combined) and a change in separate biochemical variables was calculated as RCV 1.96 x radical2 x SD(combined). Correlation between different disease activity indices was assessed for unexplained variation. The Crohn's disease activity index (CDAI) had the highest variation compared to the van Hees (Hees) and the Harvey-Bradshaw index (HBI) in CD, but it also had the best SNR, whereas HBI had the lowest. In UC the clinical activity index (CAI) showed the highest variance, but the best SNR compared to Seo's activity index (AI). The 95% CI of the CDAI discriminatory activity sum of 150 in individual cases was 105-195, whereas the 95% interval for a change was +/-62.4. Self-reported wellness contributed 40% to total variance in the CDAI. Factors of clinical importance increased errors in estimates and variance of the indices. Poor correlation was obtained between activity indices, with up to 70% unexplained variance. The SD(combined) for estimated errors was as high as 23 points, with the best SNR being approximately 20. Index factors increase the sensitivity of SNRs to errors and lower the disease specificity. Sensitivity optimisation may be achieved by standardisation of the variables and their use. PMID- 15899658 TI - An iterative method for improved estimation of the mean of peer-group distributions in proficiency testing. AB - In proficiency testing (PT), the peer-group mean is conventionally computed after twice removing values exceeding the mean +/- 3 SD. However, this adjustment fails if there are many outliers. In this study an iterative method was evaluated as a more robust way to estimate the means. The methodology repeatedly removes a proportion of the population (usually those exceeding the mean +/- 1.6 SD), assuming the presence of a Gaussian distribution in the central portion, and reinflates the SD to compensate for the trimming. A computer simulation revealed that the estimated mean of a known Gaussian distribution was less affected by a subpopulation that overlaps the main population than was the conventional method. When the overlapping portions were removed, the iterative method predicted the true mean correctly. The method was applied to external PT results for 44 analytes. Although most peer-group distributions were clearly non-Gaussian, the segment included by the predicted mean +/- 1.6 SD was regarded as Gaussian in 85.9% by the new method and 73.4% by the conventional method. The proposed methodology appears to be an improved way of estimating peer-group means. PMID- 15899659 TI - Impact of a common CV evaluation scheme on overall laboratory performance: 8-year experience of a large national proficiency testing program in Japan. AB - In proficiency testing (PT), it is difficult to set rational criteria for determining target values and allowable limits. In Japan, the largest national PT program has for the past 8 years used an evaluation scheme based on a "common coefficient of variation" (cCV) to overcome this difficulty. The cCV, the average within-method variation, is derived by one-way analysis of variance from data of peer groups after excluding those with large variations. A unit interval used for grading participants is obtained by multiplying the cCV by the peer-group mean which is computed using an iterative method featuring repeated removal of a large portion of the tails of the distribution and reinflation of the standard deviation (SD) to compensate for the trimming. The evaluation scheme gives a relatively better grade to those participants in peer groups with small CVs, compared to use of individual peer-group CVs. The scheme has expedited the transition to more reproducible methods, resulting in a notable overall improvement in laboratory performance. The cCV evaluation scheme is uniformly applicable to any analyte measured quantitatively. The iterative method, an integral part of the scheme, effectively precludes bias in the evaluation. PMID- 15899660 TI - Significance of small dense low-density lipoproteins as coronary risk factor in diabetic and non-diabetic Korean populations. AB - Small dense low-density lipoproteins (LDLs) have been associated with coronary heart disease (CHD) and type 2 diabetes in previous studies. However, the significance of small dense LDLs as a coronary risk factor for subjects with type 2 diabetes remains unclear. We measured mean LDL particle diameter by gradient gel electrophoresis (Quantimetrix Lipoprinttrade mark LDL System) in 44 type 2 diabetes patients, 100 CHD patients, 35 CHD patients with type 2 diabetes and 88 age-matched control subjects. Mean LDL particle sizes were significantly smaller (p < 0.05, Mann-Whitney test) in diabetes mellitus-only patients (25.7 +/- 0.8 nm), CHD-only patients (25.2 +/- 1.4 nm), and CHD patients with diabetes mellitus (24.9 +/- 1.6 nm) than in controls (26.2 +/- 1.4 nm). As for the mean LDL particle size, the prevalence of small dense LDLs (mean diameter < or =25.5 nm) was higher (p < 0.05, chi(2) test) in all disease groups than in controls. Furthermore, the LDL particle size in CHD patients with diabetes mellitus (24.9 nm) was significantly smaller (p = 0.049) than in diabetes mellitus-only patients (25.7 nm). Multiple logistic regression analysis revealed that small dense LDL was independently associated with the incidence of CHD in all subjects [odds ratio (OR) 4.4, 95% confidence interval (CI) 1.2-16.1, and OR 4.5, 95% CI 1.2 17.3, p < 0.05]. In this study the LDL particle size did not further decrease in CHD patients even when they had diabetes. However, the presence of CHD affects the LDL particle size in diabetes patients. These results suggest that identifying LDL size as a routine laboratory test could be used as a marker for CHD risk in both diabetic and non-diabetic Korean populations. PMID- 15899661 TI - Evaluation of a rapid bedside test for the quantitative determination of C reactive protein. AB - Regardless of its origin, any condition associated with inflammation is accompanied by an increase in serum C-reactive protein levels. This study compared the results of a rapid test for the bedside assay of C-reactive protein (QuikRead CRP, Orion Corporation, Orion Diagnostica, Espoo, Finland) with those of a standard laboratory assay in samples taken from 231 children aged less than 14 years (126 males; median age 4.7 years) attending the Emergency Department of Milan University's Institute of Pediatrics because of acute respiratory infection. The two methods showed similar median C-reactive protein levels (standard laboratory assay: 34.7 mg/L, range 4-199 mg/L; QuikRead CRP: 33.3 mg/L, range <8-196 mg/L; p = 0.779) and a similar distribution of children with C reactive protein levels of <20 mg/L, 20-70 mg/L and >70 mg/L. This study shows for the first time that the rapid QuikRead CRP test can be performed at the bedside or in an outpatient clinic and, in less than 5 min, gives the same quantitative results as those obtained using a more complex routine laboratory method. PMID- 15899662 TI - Power function of the reference change value in relation to cut-off points, reference intervals and index of individuality. AB - The reference change value, defined as RCV = 1.96 x 2(1/2) x(s(I)(2) +s A(2))(1/2), where s(I) is within-subject biological variation and s A is analytical variation, has been used for many years to take clinical decisions in patient monitoring. Furthermore, the index of individuality was defined as II = (s(I)(2) +s(A)(2))(1/2)/s(G) , where s(G) is the between-subject biological variation. This index has been simplified by later authors to s(I)/s(G) and has been used in monitoring situations to determine the utility of population-based reference intervals. Harris stated that when the index of individuality is lower than 0.6, the specific reference interval of the individual - when available - is better than the population-based reference interval. However, if a change within a patient is equivalent to the RCV applied for the significant difference between two measurements, the probability of detecting this change is only 50% (the same probability of missing it). Therefore, to obtain a higher probability of detecting a change by the RCV (e.g., 90%) the interpretation of the index of individuality has to be reconsidered. This contribution compares the power of the RCV to the use of cut-off points and population-based reference intervals. The benefits of the RCV compared to the distance to cut-off point or reference limit are also described in relation to the index of individuality. PMID- 15899663 TI - Erythrocyte ferritin concentration: analytical performance of the immunoenzymatic IMx-Ferritin (Abbott) assay. AB - Together with serum ferritin, erythrocyte ferritincan be a valuable diagnostic tool for evaluating the degree of impaired iron metabolism in different diseases. We collected peripheral blood samples from 64 subjects (22 healthy volunteers, 20 patients with hereditary hemochromatosis, and 22 patients on regular hemodialysis with secondary anemia) to evaluate whether an immunoenzymatic method generally used for serum ferritin can also be used to determine erythrocyte ferritin levels under various conditions of body iron status. Serum and erythrocyte ferritin levels were assayed in parallel using a microparticle enzyme immunoassay (MEIA) IMx-Ferritin kit and an IMx analyzer. The inter-assay imprecision of the serum and erythrocyte ferritin assays was 4.9% and 5.05%, the intra-assay imprecision was 2.2% and 2.3%, and the mean recovery was 102% (range 96-105%) and 101% (range 99-105%), respectively. Both serum and erythrocyte ferritin assays showed a detection limit of 1 microg/L and good linearity (R(2) = 0.99) in the intervals 13.9-443 and 3.9-135.6 microg/L, respectively. Our findings demonstrate that the IMx-Ferritin assay currently used to measure serum ferritin levels can also be adopted to measure erythrocyte ferritin insofar as it clearly discriminates high and low erythrocyte ferritin levels in cases of both iron overload and deficiency. PMID- 15899664 TI - Vitamin C and aberrant electrolyte results. AB - Vitamin C interferes with assays involving the redox-reaction. However, the interference of Vitamin C with electrolytes has not been reported. In the present case, we describe a 61-year-old lady with severe electrolyte abnormalities after administration of high doses of vitamin C. This patient, who had terminal colon cancer, presented to hospital with anuria. Her electrolytes were extremely abnormal (determined on the Beckman Synchron LX20): serum sodium 200 mmol/L, potassium 7.0 mmol/L, and chloride 50 mmol/L. Repeated measurements showed similar abnormalities. However, these critical abnormalities did not fit her clinical picture, as she was alert with normal vital signs. One of the specimens was also run on both the ABL700 and the Bayer644 analyzers, and the electrolytes appeared normal. Pooled serum from healthy individuals to which various amounts of vitamin C was added then was analyzed on Beckman Synchron LX20 for electrolytes, demonstrating the interference of vitamin C consistent with the initial finding. Thus, we eventually figured out that the aberrant results were due to the vitamin C caused analytical interference. PMID- 15899665 TI - POX-ACT assay and d-ROMs test: comparison impossible. PMID- 15899667 TI - Expressed genome molecular signatures of heart failure. AB - Traditional gene expression studies typically focus on one or a few genes of interest. An important limitation of single-gene studies is that they present a portrait of disease that is essentially static. However, disease is a dynamic process, driven by a combination of genetic, epigenetic and environmental factors. Recently, genomic technologies have permitted better characterization of the dynamic aspect of disease progression. Genome-wide expression profiles of cardiovascular diseases, heart failure in particular, using microarrays have been published and are providing new insights into this complex disease. Tissue biopsies required for traditional microarray studies, however, are often invasive and not readily available. By contrast, blood samples are relatively non-invasive and are readily available. In a number of recent studies, blood cells appear to be a viable substitute for tissue biopsy. Blood cells have the ability to mirror the body's tissues and organs in health and disease; thus, we hypothesize that blood cells can indicate at the molecular level the presence of disease. Here we review microarray gene expression profiling of blood RNA for a number of different diseases. Sieving through gene expression molecular signatures has identified groups of genes characteristic of each and has identified biomarkers associated with specific diseases. PMID- 15899668 TI - Understanding hyperlipidemia and atherosclerosis: lessons from genetically modified apoe and ldlr mice. AB - Hyperlipidemia is the most important risk factor for atherosclerosis, which is the major cause of cardiovascular disease. The etiology of hyperlipidemia and atherosclerosis is complex and governed by multiple interacting genes. However, mutations in two genes have been shown to be directly involved, i.e., the low density lipoprotein receptor (LDLR) and apolipoprotein E (ApoE). Genetically modified mouse models have been instrumental in elucidating the underlying molecular mechanisms in lipid metabolism. In this review, we focus on the use of two of the most widely used mouse models, ApoE- and LDLR-deficient mice. After almost a decade of applications, it is clear that each model has unique strengths and drawbacks when carrying out studies of the role of additional genes and environmental factors such as nutrition and lipid-lowering drugs. Importantly, we elaborate on mice expressing mutant forms of APOE, including the APOE3Leiden ( APOE3L ) and the APOE2 knock-in ( APOE 2k) mouse models. These models have outstanding potential, as they are highly responsive to dietary factors and pharmacological interventions. PMID- 15899669 TI - Alcohol and gene interactions. AB - Alcohol use produces both desirable and undesirable effects, ranging from short term euphoria and reduction in cardiovascular risk, to violence, accidents, dependence and liver disease. Outcomes are affected by the amount of alcohol used (which is itself affected by genetic variation) and also by the drinker's genes. Genetic effects have been most clearly demonstrated for alcohol dependence, and several of the genes for which variation leads to increased dependence risk have been identified. These include genes for enzymes involved in alcohol metabolism (alcohol dehydrogenase and aldehyde dehydrogenase), and genes for receptors affected by alcohol (particularly gamma-aminobutyric acid receptors). Many other gene/dependence associations have been reported but not fully substantiated. Genetic effects on phenotypes other than alcohol dependence are less well understood, and need to be clarified before a full picture of gene-alcohol interactions can be achieved. PMID- 15899670 TI - beta-Carotene stimulates chemotaxis of human endothelial progenitor cells. AB - Angiogenesis is a crucial process in tissue remodeling during growth, both in the embryo and the adult. In our study we concentrated on the direct effect of beta carotene on human umbilical cord originating from endothelial progenitor cells (EPCs). beta-Carotene uptake by EPCs was measured using a HPLC method. The determination of cell surface antigens was performed by flow cytometry. The effect on cell proliferation was estimated by measuring bromo-deoxyuridine incorporation. The influence on the formation of a tubular-like structure was investigated in a 3D assay in matrigel. Quantitative gene expression was estimated using real-time PCR. We demonstrated that beta-carotene in the physiological range of concentrations found in human blood is a potent activator of EPC chemotaxis, which is accompanied by a change in the expression of genes mediating cell adhesion and homing, but does not activate the final markers of endothelial differentiation. This study points to the prochemotactic and homing activity of beta-carotene in undifferentiated endothelial cell progenitors for the first time, which may suggest a potential role of this carotenoid in progenitor cell therapy aimed at angiogenesis and tissue repair. PMID- 15899671 TI - Effect of interferon-gamma, interleukin-10, lipopolysaccharide and tumor necrosis factor-alpha on chitotriosidase synthesis in human macrophages. AB - Human chitotriosidase is a chitinolytic enzyme and mainly produced by activated macrophages. Recently, we observed that prolactin, which is structurally related to several cytokines and is involved in regulating monocyte/macrophage functions, upregulates chitotriosidase gene expression in human macrophages, suggesting that chitotriosidase is not only a biochemical marker of macrophage activation in lysosomal diseases and hematological disorders, but also may reflect induction of an immunological response. To confirm this hypothesis we evaluated by quantitative real-time PCR the mRNA chitotriosidase levels in human monocytes/macrophages following treatment with pro-inflammatory stimuli such as interferon-gamma, tumor necrosis factor-alpha, lipopolysaccharide, and interleukin-10, an anti-inflammatory cytokine. Stimulation of macrophages with interferon-gamma, tumor necrosis factor-alpha and lipopolysaccharide resulted in increased levels of chitotriosidase mRNA, as well as chitotriosidase activity, whereas interleukin-10 decreased chitotriosidase synthesis. This finding is consistent with the hypothesis that the production of chitotriosidase by macrophages could have biological relevance in the immune response. PMID- 15899672 TI - Two immunochemical assays to measure advanced glycation end-products in serum from dialysis patients. AB - Advanced glycation end-products are uremic toxins that accumulate in the serum and tissues of patients with chronic renal failure. Here, we established two enzyme-linked immunosorbent assays (ELISAs) for N(epsilon)-carboxymethyllysine and imidazolone to analyze advanced glycation end-products in human serum. Both ELISAs detected advanced glycation end-products bound to human serum albumin in a dose-dependent way. Whereas the formation of imida-zolone was independent of the presence of oxygen, concentrations of N(epsilon)-carboxymethyllysine epitopes increased 20-fold under oxidative conditions. The N(epsilon)-carboxymethyllysine ELISA showed a similar response to free, peptide-bound and protein-bound N(epsilon)-carboxymethyllysine, whereas the imidazolone antibody showed slightly higher affinity toward peptide-bound compared to protein-bound imidazolone. In human serum, linear dilution ranges from 1:10 to 1:40 (N(epsilon) carboxymethyllysine ELISA) and from 1:2 to 1:8 (imidazolone ELISA) were found. The recovery of N(epsilon)-carboxymethyllysine from serum was 101 +/- 10% and 94 +/- 12%, respectively, and 93 +/- 15% and 97 +/- 12% for imidazolone. The coefficients of variation for intra-assay variability were 0.26-2.7% (N(epsilon) carboxymethyllysine) and 0.1-2.4% (imidazolone), and 8.3-13.4% (N(epsilon) carboxymethyllysine) and 7.8-12.5% (imidazolone) for inter-assay variability. In serum samples from hemodialysis patients (n = 20) and controls (n =20), an approximately two-fold increase was detected in the patient group (p < 0.001). The combination of the N(epsilon)-carboxymethyllysine and imidazolone ELISAs is a valuable tool to measure serum concentrations of advanced glycation end-products for clinical studies. PMID- 15899673 TI - Apolipoprotein E haplotyping by denaturing high-performance liquid chromatography. AB - The apolipoprotein E ( APOE ) gene in humans contains two single-base polymorphisms in exon 4, which result in three common alleles, conventionally named epsilon2 , epsilon3 and epsilon4 . Numerous studies have shown an important association between the epsilon4 variant and an increased risk of Alzheimer's disease; other data suggest a possible linkage of APOE genetic heterogeneity with lipid profile and an increased risk of atherothrombotic stroke and coronary heart disease. APOE genotyping is therefore an increasingly common assay in laboratory medicine. The most widely used technique for APOE genotyping is based upon restriction isotyping, i.e., amplification of the fragment of exon 4 containing the most common sequence variations, followed by enzymatic digestion of the amplicon and fragments analysis by gel electrophoresis. We developed a novel, reliable and fast method that exploits the sensitivity and specificity of denaturing high-performance liquid chromatography in detecting single-nucleotide polymorphisms. We show that, in most cases, with a single chromatographic separation it is possible to correctly identify the six different APOE allelic patterns, without any manipulation after the polymerase chain reaction amplification. When compared to restriction isotyping, our method is much faster, less labor intensive and similarly inexpensive. PMID- 15899674 TI - Both core and terminal glycosylation alter epitope expression in thyrotropin and introduce discordances in hormone measurements. AB - Thyroid-stimulating hormone (TSH) is routinely measured in blood to diagnose thyroid disorders using immunoassays. This study used recombinant TSH (recTSH) as a source of hormonal compound exhibiting a serum-type glycosylation and putatively reflecting physiopathological alterations in TSH polymorphism. Mass spectrometry revealed that in recTSH, both subunits display high-molecular-size glycoforms compared to the pituitary hormone (pitTSH), indicating more complex glycosylation. To determine how changes in TSH glycosylation may affect epitope expression, comparative epitope mapping of rec- and pitTSH was carried out using a panel of ten hormone-specific monoclonal antibodies. Three common epitopes, I, II and III, were identified as common to both preparations and allowed the design of six assays as I/II, II/I, I/III, III/I, II/III, and III/II. Highly sialylated recTSHs were produced by enzymatic remodeling to mimic the hormone circulating in blood and revealed limited expression of epitope I, but enhanced recognition of epitope II. Fractionation on a lentil lectin-Sepharose column allowed selection of non-fucosylated recTSH, thought to be associated with primary hypothyroidism. Recognition of epitope I was not modified by TSH core fucosylation, while epitope III expression was increased in non-fucosylated glycoforms. Taken together, our findings demonstrate that changes in both core and terminal glycosylation alter epitope expression in TSH and thereby induce highly variable antibody recognition, resulting in significant discordances among hormone measurements. PMID- 15899675 TI - Do we measure bilirubin correctly anno 2005? AB - We observed 30% discrepancy between liquid chemistry and dry chemistry analysers for the determination of total bilirubin in human adult serum samples, which were consistent with a 20% overestimation and 10% underestimation relative to a Jendrassik-Grof reference method, respectively. In contrast, standard reference material SRM916, which was recently recommended as being the most suitable material for attaining interlaboratory agreement, shows very good agreement on both types of analysers, as well as close to 100% recovery with respect to the reference method. We show that the liquid vs. dry bilirubin discrepancies seem to originate in the presence of either conjugated or delta-bilirubin. Our observations make it clear that good interlaboratory (or inter-analyser) agreement between bilirubin reference materials does not guarantee the same for bilirubin concentrations in human serum samples. PMID- 15899676 TI - Differences in mortality on the basis of laboratory parameters in an unselected population at the Emergency Department. AB - The objective of this study was to relate multiple laboratory results of initial blood gas analysis in patients visiting the emergency department to mortality. A total of 1806 adult patients visiting the emergency department for any reason, who underwent blood gas analysis, were included in this study. The results of nine laboratory parameters (ionised calcium, potassium, sodium, glucose, lactate, pH, pCO(2), pO(2) and saturation) were related to mortality. Comparison of the means of survivors' and non-survivors' age, pH, p(O)2 , glucose and lactate showed relevant differences (p < or = 0.001). When dividing the group of patients into quintiles, hypo- and hyperkalaemia (<3.6 or >4.3 mmol/L), hypocalcaemia (<1.12 mmol/L), hyperglycaemia (> 10.6 mmol/L), hyperlactataemia (>2.9 mmol/L) and acidosis (pH < 7.34) were all related to high mortality in comparison to the quintile with the lowest mortality (p < 0.0001). Interestingly, even within the reference range for lactate and ionised calcium, there was an increased risk of mortality, so these reference ranges should be interpreted with care. Glucose and lactate have additional clinical value, but are not always requested. Therefore, glucose and lactate should be routinely added to a blood gas request. Our study shows differences in mortality for laboratory parameters in patients visiting the emergency department, irrespective of the underlying pathology. PMID- 15899677 TI - An Italian program of external quality control for quantitative assays based on real-time PCR with Taq-Man probes. AB - Quantitative real-time PCR techniques are increasingly being used for the measurement of nucleic acids in research applications as well as in the clinical laboratory. It is therefore important that external quality control programs (EQA) are implemented for the evaluation of the analytical aspects common to molecular tests based on quantitative PCR. The aim of this study was the development of an Italian program of external quality control for quantitative assays based on real-time PCR with Taq-Mantrade mark probes to compare the analytical performance of 42 laboratories. Participants were provided with a set of reagents (cDNA for reference curve preparation, primers-probe mix and three unknown samples) and requested to perform a conventional assay using the master mix employed in their laboratories. The quantitative results in unknown samples were analyzed. The results of our study showed clear heterogeneity in performance. Two of the 42 laboratories provided results indicating contamination during the experiment, whereas six did not provide values for at least one of the six standard points. Only 12 laboratories gave results that were both precise and accurate for all the samples tested. Regarding imprecision, 17 laboratories appeared to deviate in at least one result, whereas inaccuracy showed an inverse dose-dependent trend. Finally, 12 laboratories were not able to measure the sample with the lowest concentration. Ten of these laboratories were equipped with the same instruments. The results of this first round of analytical EQA of real-time PCR-based methods seem to indicate high variability among laboratories carrying out the same experimental protocol. These findings could have implications for any assay based on this type of technique. This survey demonstrates the importance of experimental EQAs of methodological proficiency testing. Our approach has proved useful for comparing the analytical aspects shared by all diagnostic laboratories applying quantitative assays for the measurement of nucleic acids based on the use of Taq-Mantrade mark probes and real-time platforms. PMID- 15899678 TI - A reference material for traceability of aspartate aminotransferase (AST) results. AB - Standardization of aspartate aminotransferase (AST) determination is highly desirable for inter-laboratory comparison. Serum AST mean values for 20 patients suffering from viral hepatitis showed an inter-laboratory (n = 13) variation of 9.4%. Part of this variation was due to two laboratories using procedures without pyridoxal-5'-phosphate. A traceable AST value was assigned to an enzyme calibrator (EC) through the appropriate International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) primary reference procedure. The EC was commutable for seven routine methods. Common calibration using the EC reduced the inter-laboratory coefficient of variation (CV = 5.9% ) and allowed retention of a common reference interval for a set of routine procedures. Calibration made superfluous the expression of results in multiples of the upper reference limit, which increased inter-laboratory variation (CV = 18.5%). Furthermore, for 92% of patients, calibration with the EC allowed the correction of misclassifications when taking into account the reference interval of the reference procedure. Use of this EC could be proposed to complete the AST reference system. PMID- 15899679 TI - Harmonization of the Bayer ADVIA Centaur and Abbott AxSYM automated B-type natriuretic peptide assay in patients on hemodialysis. AB - There are two fully automated high-throughput clinical instruments for brain natriuretic peptide (BNP) assays, the Bayer ADVIA Centaur assay, and the Abbott AxSYM assay. Although both recommend a cut-off value of 100 pg/mL, we are unaware of previous studies that have compared the unadjusted results of the two methods, required for proper evaluation of patients undergoing this test on different platforms. From 43 hemodialysis patients, 80 paired samples were collected by venipuncture into plastic evacuated tubes containing EDTA. The Bayer assay yielded lower values than the Abbott assay, with linear regression of 0.53 x Abbott assay (95% confidence interval, 0.50-0.56) being forced through 0, demonstrating an r(2)-value of 0.954. Regression for the Abbott assay was 1.79 x Bayer assay (95% CI, 1.69-1.89). The cut-off values for abnormal BNP results analyzed on the Abbott system are not identical to those on the Bayer system, and this needs to be taken into account when comparing studies on the clinical utility of these systems. PMID- 15899680 TI - Multicenter evaluation of the analytical and clinical performance of the Elecsys S100 immunoassay in patients with malignant melanoma. AB - The aim of this multicenter study was to evaluate the technical and clinical performance of the Elecsys S100 electrochemiluminescence immunoassay and to assess its utility as a tumor marker in patients with malignant melanoma. Imprecision studies yielded within-run coefficients of variation (CVs) of 0.7 2.0% and between-day CVs of 1.0-6.4%. Serum samples that were distributed to 11 participating laboratories for a comparability analysis resulted in excellent recoveries of 93-105% related to the median for all laboratories. The functional sensitivity of the assay was determined to be below 0.02 microg/L. The lot-to-lot reproducibility of Elecsys S100 was tested by analyzing 110 sera with three different reagent lots on an E2010 analyzer. This lot-to-lot comparison showed excellent correlation, with a coefficient of 0.99. A 95th percentile cut-off value of 0.10 microg/L was calculated from values measured in 206 healthy individuals. Using this cut-off value, sensitivity of 41% was found, with positive and negative predictive values of 0.50 and 0.91, respectively. Method comparison with the Sangtec 100 luminescence immunoassay, run on two different analyzers, showed correlation with coefficients ranging from 0.76 to 0.95. A comparison of S100 values obtained with both tests showed identical patterns in 68 serial samples from 15 patients with malignant melanoma during follow-up. These findings indicate that serial measurements with the Elecsys S100 assay are useful for the follow-up and monitoring of therapy in patients with malignant melanoma. PMID- 15899681 TI - Guidelines for sampling, measuring and reporting ionized magnesium in undiluted serum, plasma or blood: International Federation of Clinical Chemistry and Laboratory Medicine (IFCC): IFCC Scientific Division, Committee on Point of Care Testing. AB - All analyzers with ion-selective electrodes for ionized magnesium (iMg) should yield comparable and unbiased results. The prerequisite to achieve this goal is to reach consensus on sampling, measurement and reporting. The recommended guidelines for sampling, measurement and reporting iMg in plasma ("plasma" refers to circulating plasma and the forms in which it is sampled: the plasma phase of anticoagulated whole blood, plasma separated from blood cells, or serum) or blood, referring to the substance concentration of iMg in the calibrants, will provide results for iMg that are approximately 3% greater than its true concentration, and 4% less than its true molality. Binding of magnesium to proteins and ligands in plasma and blood is pH-dependent. Therefore, pH should be simultaneously measured to allow adjustment of iMg concentration to pH 7.4. The substance concentration of iMg may be physiologically and consequently clinically more relevant than the substance concentration of total magnesium. PMID- 15899682 TI - Evaluation of the Quantase neonatal immunoreactive trypsinogen (IRT) screening assay for cystic fibrosis. PMID- 15899683 TI - A Preliminary analysis of medical expenditures among active and sedentary US adults with mental disorders. AB - OBJECTIVE: To determine whether leisure-time physical activity is associated with lower direct annual medical expenditures among a sample of adults with mental disorders. METHODS: Using the 1995 National Health Interview Survey and 1996 Medical Expenditure Panel Survey, differences between medical expenditures for sedentary and active persons were analyzed using t-tests. RESULTS: The per capita annual direct medical expenditure was US 2785 dollars higher for sedentary than for active persons (P<0.05). The total expenditure associated with sedentary behavior was US 31.7 billion dollars (US 19.1 billion dollars in men; US 12.6 billion dollars in women). CONCLUSIONS: Physical activity is associated with a reduced economic burden among people with mental disorders. PMID- 15899684 TI - Validation of the brief multidimensional students' life satisfaction scale among college students. AB - OBJECTIVE: To investigate the psychometric properties of the BMSLSS among 522 college students. METHODS: Internal consistency reliability explored scale reliability, factor analysis explored construct validity, known-groups validity was assessed by use of items from the National College Youth Risk Behavior Survey and Harvard National Survey of Alcohol Use, and criterion-related validity was explored through analyses with the CDC's Health-Related Quality of Life Scale. RESULTS: Acceptable internal consistency reliability, construct, known-groups, and criterion-related validity were established. CONCLUSIONS: These findings offer preliminary support for the BMSLSS; it could serve as part of comprehensive evaluations of subjective QOL for program evaluation and/or policy purposes among college students. PMID- 15899685 TI - Change in worksite smoking behavior following cancer risk feedback: a pilot study. AB - OBJECTIVE: To pilot a worksite smoking intervention. METHODS: Following baseline assessment, participants (N=6378) received cancer risk feedback; 2 annual evaluations were conducted. RESULTS: Using all data, smoking dropped from 13.7% to 8.4% and 9.3%, and smoker's readiness to quit increased. Using complete data, smoking initially increased from 5.7% to 6.7%, but subsequently decreased to 5.3%; the increase in smoker's readiness to quit remained. Being male, younger, and with lower education and self-efficacy predicted smoking. Lower age and higher self-efficacy predicted readiness to quit smoking. CONCLUSIONS: These findings support a formal evaluation of a worksite smoking intervention using cancer risk feedback. PMID- 15899686 TI - Evaluation of an intervention for hospitalized African American smokers. AB - OBJECTIVE: To evaluate the effectiveness of a smoking cessation intervention based on the transtheoretical model of change with a sample of low-income African American smokers admitted to an indigent-care hospital. METHODS: The intervention incorporated components shown to be effective in increasing cessation in other populations, tailored to a bedside counseling format with follow-up contact postdischarge. RESULTS: Intervention patients were significantly more likely to advance in stage than were control patients. CONCLUSION: A hospital-offered bedside intervention offers promise in reaching underserved smokers with effective, though limited, cessation assistance. PMID- 15899687 TI - Bupropion SR vs placebo for smoking cessation in health care professionals. AB - OBJECTIVE: To evaluate bupropion SR for smoking cessation in physicians and nurses. METHODS: This double-blind prospective 26-center, 12-country trial randomized 687 subjects to smoking cessation counselling with bupropion SR or placebo for 7 weeks. The participants were followed for 52 weeks. RESULTS: Bupropion SR was superior to placebo (50% vs 40%, P=0.013) on the 4-week primary outcome variable. Due to a high placebo response in this health care population, statistical differences were not maintained after treatment was discontinued. CONCLUSIONS: Bupropion SR is effective and well tolerated in health care professionals. Relapse prevention measures are needed to attain long-term abstinence. PMID- 15899689 TI - Norms and their relationship to behavior in worksite settings: an application of the Jackson Return Potential Model. AB - OBJECTIVES: To measure health norms and assess their influence on behavior among 2541 employees in 16 manufacturing worksites using an adapted Jackson's Return Potential Model (RPM). METHODS: Worksite-level norm intensity, crystallization, and normative power were calculated for several behaviors; linear regression analyses tested whether normative power was related to each health behavior. RESULTS: Norms about safe work practices and smoking were most intense; norms about safe work practices were most crystallized. Safe work practices and smoking held the highest normative power; healthy eating held the least normative power. CONCLUSIONS: Comparing norm characteristics across health behaviors leads to important leverage points for intervening to influence norms and improve worker health. PMID- 15899688 TI - Scale development for perceived school climate for girls' physical activity. AB - OBJECTIVES: To test an original scale assessing perceived school climate for girls' physical activity in middle school girls. METHODS: Confirmatory factor analysis (CFA) and structural equation modeling (SEM). RESULTS: CFA retained 5 of 14 original items. A model with 2 correlated factors, perceptions about teachers' and boys' behaviors, respectively, fit the data well in both sixth and eighth graders. SEM detected a positive, significant direct association of the teacher factor, but not the boy factor, with girls' self-reported physical activity. CONCLUSIONS: School climate for girls' physical activity is a measurable construct, and preliminary evidence suggests a relationship with physical activity. PMID- 15899690 TI - ViewPoint: conversation with John P. Elder, PhD. PMID- 15899692 TI - Nicking activity on pBR322 DNA of ribosome inactivating proteins from Phytolacca dioica L. leaves. AB - Ribosome-inactivating proteins isolated from Phytolacca dioica L. leaves are rRNA N-glycosidases, as well as adenine polynucleotide glycosylases. Here we report that some of them cleave supercoiled pBR322 dsDNA, generating relaxed and linear molecules. PD-L1, the glycosylated major form isolated from the winter leaves of adult P . dioica plants, produces both free 3'-OH and 5'-P termini randomly distributed along the DNA molecule, as suggested by labelling experiments with [alpha- 32P]dCTP and [gamma- 32 P]dATP. Moreover, when the reaction is carried out under low-salt conditions, cleavage is observed mainly at a specific site, located downstream of the ampicillin resistance gene (close to position 3200), ending with the deletion of a fragment of approximately 70 nucleotides. This cleavage pattern is similar to that obtained under the same conditions with mung bean nuclease, a single-strand endonuclease. Furthermore, pBR322 DNA treated with PD-L1 shows reduced transforming activity with E . coli HB101 competent cells in comparison to untreated control plasmid DNA. PMID- 15899693 TI - Identification of three novel mutations in the dihydropyrimidine dehydrogenase gene associated with altered pre-mRNA splicing or protein function. AB - Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in the catabolism of the pyrimidine bases uracil and thymine, as well as of the widely used chemotherapeutic drug 5-fluorouracil (5FU). Analysis of the DPD gene ( DPYD ) in two patients presenting with complete DPD deficiency and the parents of an affected child showed the presence of three novel mutations, including one splice site mutation IVS11 + 1G-->T and the missense mutations 731A-->C (E244V) and 1651G-->A (A551T). The G-->T mutation in the invariant GT splice donor site flanking exon 11 (IVS11 + 1G-->T) created a cryptic splice site within exon 11. As a consequence, a 141-bp fragment encoding the aminoacid residues 400-446 of the primary sequence of the DPD protein was missing in the mature DPD mRNA. Analysis of the crystal structure of pig DPD suggested that the E244V mutation might interfere with the electron flow between NADPH and the pyrimidine binding site of DPD. The A551T point mutation might prevent binding of the prosthetic group FMN and affect folding of the DPD protein. The identification of these novel mutations in DPYD will allow the identification of patients with an increased risk of developing severe 5FU-associated toxicity. PMID- 15899694 TI - The connexin gene family in mammals. AB - Unannotated mammalian genome databases (dog, cow, opossum) were searched for candidate connexin genes, using sequences from annotated genomes (man, mouse). All 18 'multi-species' connexin genes, i.e., orthologs of connexin26 , 29/31.3 (duplicated in opossum), 30, 30.2/31.9, 30.3, 31, 31.1, 32, 36, 37, 39/40.1, 40, 43, 45, 44/46, 47, 50, and 57/62 , were found in dog, cow and opossum. Connexin25 and 58 have been considered specific for man, but evident orthologs of connexin25 were found in dog, cow and opossum, and orthologs of connexin58 were found in dog and cow. Moreover, a connexin43 -like sequence (approx. 80% identical to connexin43 ) was found in man, chimpanzee, dog and cow. In the three former species, the sequences were located on the X chromosome. In man, chimpanzee and cow, there were stop codons in all reading frames; these sequences are therefore judged as pseudogenes, called here Cx43pX . In the dog, the sequence contained an open reading frame for a protein of 35.7 kDa (connexin35.7). We suggest that these sequences are orthologs of connexin33 , previously considered as a rodent specific connexin gene. Thus, connexin25 , 33 and 58 are not species-specific genes. However, the opossum may possess a candidate, connexin39.2 , without obvious orthologs in other mammals. Furthermore, pseudogenes of primate connexin31.3 and opossum connexin35 (one of the two orthologs of primate connexin31.3) were detected. These results suggest that the structure of the mammalian connexin gene family should be revised, especially with regard to the so-called 'species-specific' connexins . PMID- 15899695 TI - Hydrogen peroxide causes greater oxidation in cellular RNA than in DNA. AB - Human A549 lung epithelial cells were challenged with 18O-labeled hydrogen peroxide ([18O]-H2O2), the total RNA and DNA extracted in parallel, and analyzed for 18O-labeled 8-oxo-7,8-dihydroguanosine ([18O]-8-oxoGuo) and 8-oxo-7,8-dihydro 2'-deoxyguanosine ([18O]-8-oxodGuo) respectively, using high-performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-MS/MS). [18O]-H2O2 exposure resulted in dose-response formation of both [18O]-8-oxoGuo and [18O]-8-oxodGuo and 18O-labeling of guanine in RNA was 14-25 times more common than in DNA. Kinetics of formation and subsequent removal of oxidized nucleic acids adducts were also monitored up to 24 h. The A549 showed slow turnover rates of adducts in RNA and DNA giving half-lives of approximately 12.5 h for [18O]-8-oxoGuo in RNA and 20.7 h for [18O]-8-oxodGuo in DNA, respectively. PMID- 15899696 TI - Homology modeling and SAR analysis of Schistosoma japonicum cathepsin D (SjCD) with statin inhibitors identify a unique active site steric barrier with potential for the design of specific inhibitors. AB - Proteases that digest the blood-meal of the parasitic fluke Schistosoma are potential targets for therapy of schistosomiasis, a disease of chronic morbidity in humans. We generated a three-dimensional model of the cathepsin D target protease of Schistosoma japonicum (SjCD) utilizing the crystal structure of human cathepsin D (huCD) in complex with pepstatin as template. A homology model was also generated for the related secreted aspartic protease 2 (SAP2) of the pathogenic yeast, Candida albicans . An initial panel of seven statin inhibitors, originally designed for huCD [Majer et al., Protein Sci. 6 (1997), pp. 1458 1466], was tested against the two pathogen proteases. One inhibitor showed poor reactivity with SjCD. Examination of the SjCD active-site cleft revealed that the poor inhibition was due to a unique steric barrier situated between the S2 and S4 subsites. An in silico screen of 20 potential statin scaffolds with the SjCD model and incorporating the steric barrier constraint was performed. Four inhibitors (SJ1-SJ4) were eventually synthesized and tested with SjCD, bovine CD and SAP2. Of these, SJ2 and SJ3 proved moderately more specific for SjCD over bovine CD, with IC 50 values of 15 and 60 nM, respectively. The unique steric barrier identified here provides a structural focus for further development of more specific SjCD inhibitors. PMID- 15899697 TI - Interpretation of the reactivity of peroxidase compound II with phenols and anilines using the Marcus equation. AB - The catalytic cycle of heme peroxidases involves three processes: the formation of compound I, its conversion to compound II and regeneration of the native enzyme. Each of the processes consists of a reversible binding stage followed by an irreversible transformation stage. Our group has proposed a continuous, sensitive and reliable chronometric method for measuring the steady-state rate of peroxidase activity. Furthermore, we have derived an analytical expression for the steady-state rate and simplified it, taking into consideration the experimental values of the rate constants of some stages previously determined by other authors in stopped-flow assays. We determined the value of the constant for the transformation of a series of phenols and anilines by compound II, and found that it involves a deprotonation step and an electron transfer step. Study of the solvent deuterium isotope effect on the oxidation of phenol revealed the non-rate limiting character of the deprotonation step in a proton inventory study. Usage of the Marcus equation showed that the electronic transfer step is rate-limiting in both cases, while phenols and anilines were oxidised at different rates for the same potentials. This can be attributed to the shorter electron-tunnelling distance for electron transfer to the iron ion in the phenols than in the anilines. PMID- 15899698 TI - P. falciparum pro-histoaspartic protease (proHAP) protein peptides bind specifically to erythrocytes and inhibit the invasion process in vitro. AB - Plasmodium falciparum histoaspartic protease (HAP) is an active enzyme involved in haemoglobin degradation. HAP is expressed as an inactive 51-kDa zymogen and is cleaved into an active 37-kDa enzyme. It has been proposed that this kind of protease might be implicated in the parasite's invasion of erythrocytes; however, this protein's role during invasion has still to be determined. Synthetic peptides derived from the HAP precursor (proHAP) were tested in erythrocyte binding assays to identify their possible function in the invasion process. Two proHAP high-activity binding peptides (HABPs) specifically bound to erythrocytes; these peptides were numbered 30609 (101LKNYIKESVKLFNKGLTKKS120) and 30610 (121YLGSEFDNVELKDLANVLSF140 ). The binding of these two peptides was saturable, presenting nanomolar affinity constants. These peptides interacted with 26- and 45-kDa proteins on the erythrocyte surface; the nature of these receptor sites was studied in peptide binding assays using enzyme-treated erythrocytes. The HABPs showed greater than 90% merozoite invasion inhibition in in vitro assays. Goat serum containing proHAP polymeric peptide antibodies inhibited parasite invasion in vitro . PMID- 15899699 TI - The snake venom metalloproteases berythractivase and jararhagin activate endothelial cells. AB - PIII snake venom metalloproteases (SVMPs) are metalloproteases structurally related to ADAMs (a disintegrin and metalloprotease human family of proteins). Berythractivase and jararhagin are PIII SVMPs with 69% homology that have different hemostatic properties. In order to clarify these differences and further characterize the biological effects of these proteins, we have analyzed the effect of both proteases on human umbilical-vein endothelial cell functions. We found that both proteins enhanced nitric oxide generation, prostacyclin production and interleukin-8 release. Berythractivase but not jararhagin increased the expression of decay accelerating factor. Jararhagin decreased cell viability in a concentration-dependent manner and induced cellular apoptosis, while berythractivase did not modulate cell survival. Our results show for the first time that, besides the known anti-aggregating or procoagulant effects of PIII SVMPs, these proteins trigger endothelial cell effector responses. Although structurally related, berythractivase and jararhagin induce a dissimilar generation and release of endothelial molecules that may account for their different hemorrhagic activity. PMID- 15899700 TI - Visualisation of transforming growth factor-beta 1, tissue kallikrein, and kinin and transforming growth factor-beta receptors on human clear-cell renal carcinoma cells. AB - Transforming growth factor-beta1 (TGF-beta1) has a biphasic effect on the growth of renal epithelial cells. In transformed cells, TGF-beta1 appears to accelerate the proliferation of malignant cells. The diverse cellular functions of TGF-beta1 are regulated by three high-affinity serine/threonine kinase receptors, namely TbetaRI, TbetaRII and TbetaRIII. The renal serine protease tissue kallikrein acts on its endogenous protein substrate kininogen to form kinin peptides. The cellular actions of kinins are mediated through B1 and B2 G protein-coupled rhodopsin receptors. Both kinin peptides and TGF-beta1 are mitogenic, and therefore may play an important role in carcinogenesis. Experiments were designed to immunolabel tissue kallikrein, TGF-beta1, TbetaRII, TbetaRIII and kinin receptors using specific antibodies on serial sections of normal kidney and clear cell renal carcinoma (CCRC) tissue, which included both the tumour and the adjacent renal parenchyma. The essential result was the localisation of tissue kallikrein, kinin B 1 and B 2 receptors and TGF-beta1 primarily on the cell membranes of CCRC cells. In the distal and proximal tubules of the renal parenchyma adjacent to the carcinoma (RPTAC), immunolabelling for tissue kallikrein was reduced, but the expression of kinin B1 and B2 receptors was enhanced. Immunolabelling for TbetaRII and TbetaRIII was more pronounced in the proximal tubules of the tissue adjacent to the carcinoma when compared to the normal kidney. The expression of tissue kallikrein, kinin receptors, and TbetaRII and TbetaRIII may be relevant to the parenchymal invasion and metastasis of clear cell renal carcinoma. PMID- 15899701 TI - cDNA cloning and heterologous expression of a wheat proteinase inhibitor of subtilisin and chymotrypsin (WSCI) that interferes with digestive enzymes of insect pests. AB - A cDNA encoding the proteinase inhibitor WSCI (wheat subtilisin/chymotrypsin inhibitor) was isolated by RT-PCR. Degenerate oligonucleotide primers were designed based on the amino acid sequence of WSCI and on the nucleotide sequence of the two homologous inhibitors (CI-2A and CI-2B) isolated from barley. For large-scale production, wsci cDNA was cloned into the E. coli vector pGEX-2T. The fusion protein GST-WSCI was efficiently produced in the bacterial expression system and, as the native inhibitor, was capable of inhibiting bacterial subtilisin, mammalian chymotrypsins and chymotrypsin-like activities present in crude extracts of a number of insect larvae ( Helicoverpa armigera , Plodia interpunctella and Tenebrio molitor ). The recombinant protein produced was also able to interfere with chymotrypsin-like activity isolated from immature wheat caryopses. These findings support a physiological role for this inhibitor during grain maturation. PMID- 15899702 TI - Proteolytic susceptibility of the serine protease inhibitor trappin-2 (pre elafin): evidence for tryptase-mediated generation of elafin. AB - A number of serine, cysteine, metallo- and acid proteases were evaluated for their ability to proteolytically cleave the serine protease inhibitor trappin-2, also known as pre-elafin, and to release elafin from its precursor. None of the metalloproteases or acid proteases examined cleaved trappin-2, while serine and cysteine proteases preferentially cleaved trappin-2 within its non-inhibitory N terminal moiety. Cathepsin L, cathepsin K, plasmin, trypsin and tryptase were able to release elafin by cleaving the Lys 38 -Ala 39 peptide bond in trappin-2. However, purified tryptase appeared to be efficient at releasing elafin. Incubation of trappin-2 with purified mast cells first challenged with anti immunoglobulin E or calcium ionophore A23187 resulted in the rapid generation of elafin. This proteolytic release of elafin from trappin-2 was inhibited in the presence of a tryptase inhibitor, suggesting that this mast cell enzyme was involved in the process. Finally, ex vivo incubation of trappin-2 with sputum from cystic fibrosis patients indicated the production of a proteolytic immunoreactive fragment with the same mass as that of native elafin. This cleavage did not occur when preincubating the sputum with polyclonal antibodies directed against tryptase. Taken together, these findings indicate that tryptase could likely be involved in the maturation of trappin-2 into elafin under physiological conditions. PMID- 15899703 TI - Labelling of four distinct trophozoite falcipains of Plasmodium falciparum by a cystatin-derived probe. AB - Trophozoite cysteine protease (TCP) activity, isolated from Plasmodium falciparum soluble 100,000 g extracts, displayed native falcipain-1 kinetic parameters towards peptidyl substrates. The labelling of either isolated TCP or soluble 100,000 g extracts by a cystatin-derived probe (biotinyl-Leu-Val-Gly-CHN2) revealed a single band of ca. 30 kDa by SDS-PAGE, which was resolved into four spots displaying isoelectric points (pI) from 4.7 to 5.3 after two-dimensional separation. The molecular mass and pI correspond to those of falcipain-3, falcipain-2, falcipain-2' and falcipain-1, respectively. The two central spots were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry as falcipain-2 and falcipain-2'. This activity-based probe represents a potential tool for profiling active falcipains in parasites. PMID- 15899704 TI - Parallels between aging and schizophrenia. AB - As I grow old and struggle to cope with the infirmities of age, I marvel at the resilience of my patients who grapple with and overcome the indignities of schizophrenia. Schizophrenia research is mainly about the deficits, about the many difficulties patients face. It rarely celebrates the accomplishments of patients, their ability, on the whole, to accommodate and adapt to the constraints of their illness. This paper focuses on the gerontology literature, where old age is conceptualized less as defeat, more as a process one can successfully manage. Effective strategies old people use are studied and valued. This is in marked contrast with the psychiatric literature that pathologizes the compensatory strategies used in schizophrenia. PMID- 15899705 TI - Crystallized intelligence versus fluid intelligence. PMID- 15899706 TI - Why some patients succeed. PMID- 15899707 TI - A family approach to severe mental illness in post-war Kosovo. AB - This study describes the effects of a psychoeducational multiple-family group program for families of people with severe mental illness in post-war Kosovo that was developed by a Kosovar-American professional collaborative. The subjects were 30 families of people with severe mental illnesses living in two cities in Kosovo. All subjects participated in multiple-family groups and received family home visits. The program documented medication compliance, number of psychiatric hospitalizations, family mental health services use, and several other characteristics, for the year prior to the groups and the first year of the groups. The families attended an average of 5.5 (out of 7) groups, and 93% of these families attended four or more meetings. The uncontrolled pre- to post intervention comparison demonstrated decreases in medication non-compliance and hospitalizations, and increases in family mental health service use. The program provided training for mental health professionals, led to policy change in the Ministry of Health, and resulted in dissemination to other community mental health centers. This study provides preliminary evidence that a collaboratively designed and implemented psychoeducational, multiple-family program is a feasible and beneficial intervention for families of people with severe mental illness in impoverished post-war settings. PMID- 15899708 TI - Media exposure to bioterrorism: stress and the anthrax attacks. AB - This study examined media exposure and adjustment to anthrax bioterrorism attacks and the terrorist attacks on 9/11 in a sample of 300 people who lived distant from the attacks. Measures of direct and indirect exposure to terrorism, perceived risk of anthrax exposure, psychological distress, and outlook were assessed at 2 to 3 months and at 8 months after the first reported anthrax attack. Initial anthrax media exposure was a powerful predictor of distress, whereas subsequent anthrax media exposure only predicted negative changes in outlook over time. Perceived risk of anthrax exposure predicted distress and outlook but did not appear to mediate the effects of media exposure. Determining the nature and consequences of media exposure to threatening and frightening events like terrorism will help predict and manage response to future bioterrorism. PMID- 15899709 TI - The process of change in Interpersonal Psychotherapy (IPT) for depression: a case study for the new IPT therapist. AB - This case study involves a set of transcripts of Interpersonal Psychotherapy (IPT) sessions from the Christchurch Psychotherapy for Depression Study. The case study explores the techniques employed by the therapist for a depressed patient who has identified interpersonal disputes as the interpersonal area to focus on. The psychotherapeutic interventions utilized by the IPT therapist included: seeking information; exploring parallels in other relationships; exploring relationship patterns; exploring communication patterns; signalling what is significant; providing support, exploring affect; exploring options; problem solving; drawing analogy; and challenging. The themes that emerged in the patient's response to IPT were: struggling, deconstructing, connecting, practicing and reconstructing. PMID- 15899710 TI - Treating attachment injured couples with emotionally focused therapy: a case study approach. AB - This paper compared the attachment injury resolution process in two distressed couples undergoing ten sessions of Emotional Focused Therapy (EFT), a short-term empirically validated treatment for relational distress. An attachment injury is a newly coined clinical construct that denotes a specific type of betrayal within the couple's relationship. The incident is so potent that it calls into question assumptions about the safety of the relationship. The task analytic method was used to examine the pathways of change as related to attachment injury of each couple. Several outcome and process measures were employed in order to differentiate the therapeutic process between the resolved versus non-resolved couple. Results indicated that the couple who resolved their identified attachment injury at the outset of therapy adhered to the attachment injury resolution model, while the non-resolved couple showed marked deviations from the expected pathways of change. Findings suggest that the resolved couple tended to show more differentiation of interactional positions and greater levels of experiencing throughout the therapeutic process in relation to the non-resolved couple. It is recommended that further research is necessary to examine the clinical utility of the attachment injury resolution model in the context of a larger number of case studies. PMID- 15899712 TI - Alternative dimensional models of personality disorder: finding a common ground. AB - The recognition of the many limitations of the categorical model of personality disorder classification has led to the development of quite a number of alternative proposals for a dimensional classification. The purpose of this article is to suggest that future research work toward the integration of these alternative proposals within a common hierarchical structure. An illustration of a potential integration is provided using the constructs assessed within existing dimensional models. Suggestions for future research that will help lead toward a common, integrative dimensional model of personality disorder are provided. PMID- 15899713 TI - Behavioral and molecular genetic contributions to a dimensional classification of personality disorder. AB - This article examines the possible contribution of behavioral and molecular genetic research to the development of a dimensional classification of personality disorder. It is argued that the results of molecular studies are too preliminary to have immediate nosological significance. However, behavioral genetic methods could play a useful role in constructing a classification that reflects the genetic architecture of personality disorder. It is also argued that the best approach to constructing a valid classification would be to integrate behavioral genetic methods with the construct validation framework used in test construction. An integrative approach is proposed that seeks to combine constructs from alternative dimensional models. It is suggested that strong evidence of a four-dimensional structure to personality disorder provides a way to organize a preliminary model. An initial set of primary traits to define these secondary domains would then be compiled from existing models and refined using a combination of traditional psychometric analyses and behavioral genetic methods. It is concluded that an etiologically based classification is feasible for the DSM-V. PMID- 15899714 TI - Neurobiological dimensional models of personality: a review of the models of Cloninger, Depue, and Siever. AB - Neurobiological dimensional models of personality aim to account for the structure of personality traits and disorders through links to neurotransmitter systems. Three such models are reviewed: those of Cloninger, Depue, and Siever. While these proposals have heuristic value, none of them has obtained strong empirical support. Our current understanding of neurobiology is insufficient to develop a model of personality on this basis. At this point, dimensions of personality should be derived from factor analysis rather than from neurobiological theories. PMID- 15899715 TI - Temperament, personality, and developmental psychopathology as childhood antecedents of personality disorders. AB - To contribute to the case for a dimensional conceptualization of psychopathology in general and maladaptive personality or personality disorders in particular, the present paper reviews the evidence for a dimensional representation of childhood temperament and personality. The review of temperament and variable centered as well as person-centered approaches to childhood personality leads us to propose five broadband dimensions that capture individual differences in children and adolescents: extraversion, emotional stability, agreeableness, conscientiousness, and openness/intellect. Our analysis of the CBCL (Achenbach, 1991) and the DIPSI (currently under development at Ghent University), two dimensional models for childhood psychopathology, suggests two common broadband factors, internalizing and externalizing. The relations between the dimensional representation of childhood temperament/personality and psychopathology are documented with data from general population and clinical samples of children and adolescents. The article concludes with a proposal on how the higher-order dimensions emerging from studies of adaptive and maladaptive individual differences in childhood could be integrated in a common dimensional model. PMID- 15899716 TI - A developmental perspective on personality disorders: lessons from research on normal personality development in childhood and adolescence. AB - Recent work on normal personality development in children and adolescents points to several conclusions that are relevant for understanding personality pathology. First, child temperament and adult personality traits share many features in common. Second, youths' individual differences can be described in terms of the Big Five personality traits observed in adults; an integrative taxonomy of individual differences in childhood and adolescence is articulated in this article. Third, personality is already moderately stable by the preschool years, but considerable personality change occurs well into the adult years. Taken together, these findings suggest that childhood personality functioning can and should be integrated into developmental research and applied work on personality disorders. PMID- 15899717 TI - An investigation of organic factors in the neuropsychological functioning of patients with borderline personality disorder. AB - The hypothesis to be tested in this study was that the cognitive deficits that have been documented in patients with Borderline Personality Disorder (BPD) are largely the consequence of organic insult, either developmental or acquired. Using a cross-sectional design, 80 subjects (males and females) who met the criteria for BPD participated in the study. They completed a battery of neuropsychological tests and a comprehensive interview assessing organic status as well as measures of the potentially confounding factors of current levels of depression and anxiety. It was expected that BPD-patients with a probable history of organic insult would perform significantly worse than would BPD patients without such a history. Analyses of the results provided partial support for the hypothesis. Subjects with both BPD and a history of organic insult were significantly more impaired on several measures including measures of attention than were BPD only subjects. The results suggested that the impaired cognitive performance of persons diagnosed with BPD may, in part, be attributed to organic factors. PMID- 15899718 TI - Psychosocial functioning of borderline patients and axis II comparison subjects followed prospectively for six years. AB - The purpose of this study was to determine the course of the psychosocial functioning of patients with borderline personality disorder (BPD) over 6 years of prospective follow-up. The psychosocial functioning of 290 patients meeting both DIB-R and DSM-III-R criteria for BPD and 72 patients meeting DSM-III-R criteria for another Axis II disorder (and neither criteria set for BPD) was assessed at baseline using a semistructured interview of demonstrated reliability. Over 94% of surviving patients were reinterviewed about their psychosocial functioning blind to all previously collected information at three distinct follow-up waves: 2-, 4-, and 6-year follow-up. The psychosocial functioning of borderline patients improved substantially over time, with the percentage meeting criteria for good overall psychosocial functioning increasing from 26% at baseline to 56% during the third wave of follow-up. Despite this improvement, borderline patients functioned significantly more poorly than Axis II comparison subjects, particularly in the area of vocational achievement. However, a more detailed examination revealed that borderline patients who had experienced a symptomatic remission during the course of the study functioned significantly better both socially and vocationally than never-remitted borderline patients. More specifically, they were significantly more likely to have a good relationship with a spouse/partner and at least one parent, good work/school performance, a sustained work/school history, a GAF score of 61 or higher (43% vs. 0% 6 years after their index admission), and to have good overall psychosocial functioning (66% vs. 27% at 6 year follow-up). Taken together, the results of this study suggest that psychosocial improvement is both common among borderline patients and strongly related to their symptomatic status. PMID- 15899719 TI - Self-reported personality disorder in the children in the community sample: convergent and prospective validity in late adolescence and adulthood. AB - Approximately 800 youths from the Children in the Community Study (Cohen & Cohen, 1996) have been assessed prospectively for over 20 years to study personality disorders (PDs) in adolescents and young adults. In this article we evaluate the Children in the Community Self-Report (CIC-SR) Scales, which were designed to assess DSM-IV PDs using self-reported prospective data from this longitudinal sample. To evaluate convergent validity, we assessed concordance between the CIC SR Scales and the Structured Clinical Interview for DSM-IV Personality Disorders (SCID-II; First, Gibbon, Spitzer, Williams, & Benjamin, 1995) in 644 participants at mean age 33. To assess predictive validity, we used CIC-SR Scales at mean age 22 to predict subsequent CIC-SR and SCID-II Personality Questionnaire scores at mean age 33. In these analyses the CIC-SR Scales matched or exceeded benchmarks established in previous comparisons between self-report instruments and structured clinical interviews. Unlike other self-report scales, the CIC-SR did not appear to overestimate diagnoses when compared with SCID-II clinical diagnoses. PMID- 15899720 TI - Dimensional personality traits and the prediction of DSM-IV personality disorder symptom counts in a nonclinical sample. AB - The third edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-III; APA, 1980) set forth a categorical system of personality psychopathology that is composed of discrete personality disorders (PDs), each with a distinct set of diagnostic criteria. Although this system is widely accepted and highly influential, alternative dimensional approaches to capturing personality psychopathology have been proposed. Three dimensional models of personality have garnered particular attention-the Five-Factor Model (FFM; Costa & McCrae, 1992), the Seven-Factor Psychobiological Model of Temperament and Character (Seven-Factor Model; Cloninger, Svrakic, & Przybeck, 1993); and the 18 factor model of personality pathology (18-factor model; Livesley, 1986). Although the personality traits from each of these models has been examined in relation to the ten personality disorders in the DSM-IV, no study has examined the comparative and incremental validity of these models in predicting PD symptoms for these ten disorders. Using self-report instruments that measure these models and the ten DSM-IV PDs, correlation and linear regression analyses indicate that traits from all three models had statistically significant associations with PD symptom counts. Hierarchical regressions revealed that the 18-factor model had incremental predictive validity over the FFM and Seven-Fac-tor Model in predicting symptom counts for all ten DSM-IV PDs. The FFM had incremental predictive validity over the Seven-Factor Model model for all ten disorders and the Seven-Factor was able to add incremental predictive validity over the 18 factor model for five of the ten PDs and for eight of the ten disorders relative to the FFM. PMID- 15899721 TI - The impact of parental borderline personality disorder on vulnerability to depression in children of affectively ill parents. AB - Children of parents with major depressive disorder (MDD) are four to six times more likely than other children to develop MDD. Little research has examined whether comorbid parental diagnoses further increase children's risk. This study examines whether children of parents with comorbid MDD and Borderline Personality Disorder (BPD) (1) are at greater risk for experiencing depressive symptoms and/or episodes and (2) whether such increased risk may be due, in part, to their exhibiting higher levels of cognitive/interpersonal vulnerability factors. Children (n = 140; ages 6-14) of parents with MDD completed measures assessing cognitive/interpersonal vulnerability factors. Parents completed semi-structured clinical interviews assessing severity of current depressive symptoms and BPD. Both children and parents completed a semi-structured clinical interview assessing the child's current and past history of MDD. Children of parents with comorbid MDD and BPD exhibited higher levels of current depressive symptoms and higher levels of cognitive/interpersonal vulnerability factors than children of parents with MDD but no BPD, even after controlling for parents' current levels of depressive symptoms. The relationship between parental BPD and chil-dren's current levels of depressive symptoms was partially mediated by children's cognitive/interpersonal vulnerability factors. Last, children of parents with comorbid BPD and MDD were 6.84 times more likely to exhibit a current or past diagnosis of MDD. PMID- 15899722 TI - Personality disorders in patients with burning mouth syndrome. AB - Burning Mouth Syndrome (BMS) presents high rates of comorbid Axis I disorders while no controlled studies have addressed the question of Axis II comorbidities. The aim of the present study was to examine DSM-IV (APA, 1994) Axis II comorbidity in BMS patients and to control for the specificity of this association. Seventy BMS patients were compared to a nonpsychiatric population sample and to patients with other Somatoform Disorders for the presence of personality disorders (assessed with the Structured Clinical Interview for DSM-IV Axis II Personality Disorders [SCID-II; First, Gibbon, Spitzer, & Williams, 1997). Prevalence rates were compared using the Pearson's chi square test. At least one personality disorder (PD) was found in 85.7%, 24.3%, and 88.6% of subjects in the three groups, respectively. When examining PD subgroups, significant differences emerged even between the BMS and the somatoform disorder group, with BMS patients showing more Cluster A and fewer Cluster B PDs. Our results suggest that BMS is associated with a specific pattern of Axis II comorbidity. PMID- 15899723 TI - Linking adult psychopathy with childhood hyperactivity-impulsivity-attention problems and conduct problems through retrospective self-reports. AB - The purpose of the present study was to test whether adult criminals with psychopathy diagnoses, more than those without, have histories of hyperactivity impulsivity-attention problems (HIA) and conduct problems (CP). We compared psychopathic and nonpsychopathic violent criminal offenders on retrospective reports of conduct problems before the age of 15 and hyperactivity-impulsivity attention problems before the age of 10. We used a sample of 186 adult men sentenced to prison in Sweden for 4 years or more for violent, nonsexual crimes. The mean age was 30.7( SD = 9.4). The results showed that a combination of childhood HIA problems and CP was typical for adult psychopathic offenders. They were four times more likely than chance to have had a combination of HIA problems and CP during childhood and only one-fifth as likely than chance to have had neither problem. Nonpsychopathic offenders, on the other hand, were five times more likely than chance to have had neither problem and only one-quarter as likely than chance to have had both problems. PMID- 15899724 TI - Antithrombotic effects of rosiglitazone-metformin versus glimepiride-metformin combination therapy in patients with type 2 diabetes mellitus and metabolic syndrome. AB - STUDY OBJECTIVE: To evaluate the differential effect on coagulation and fibrinolysis parameters of combination therapy with glimepiride-metformin and with rosiglitazone-metformin beyond their effect on glucose metabolism in patients with type 2 diabetes and metabolic syndrome. DESIGN: Multicenter, double blind, randomized, controlled trial. SETTING: Two university-affiliated medical centers in Italy. PATIENTS: Ninety-five patients with type 2 diabetes for at least 6 months without glycemic control by diet and oral hypoglycemic agents to their maximum tolerated dosage and who also had metabolic syndrome. INTERVENTION: All 95 patients received metformin 1500 mg/day. In a randomized manner, 47 patients received glimepiride 2 mg/day and 48 patients received rosiglitazone 4 mg/day. MEASUREMENTS AND MAIN RESULTS: Body mass index (BMI), glycemic control, and coagulation and fibrinolysis parameters were evaluated at 3, 6, 9, and 12 months of treatment. Compared with baseline values, significant decreases in BMI, fasting plasma glucose, postprandial plasma glucose, and hemoglobin A1c were observed at 12 months in both the glimepiride and rosiglitazone groups (p<0.05 and p<0.01, respectively). Decreases in fasting plasma insulin and postprandial plasma insulin were observed at 12 months (p<0.05 and p<0.01, respectively) compared with baseline values in the rosiglitazone group. Furthermore, improvement in the Homeostasis Model Assessment index was observed only at 9 and 12 months (p<0.05 and p<0.01, respectively) compared with baseline in the rosiglitazone group. Significant improvement in plasminogen activator inhibitor (PAI)-1 was present in the rosiglitazone group after 9 months (p<0.05), and significant PAI-1 improvement was observed in the glimepiride and rosiglitazone groups after 12 months (p<0.05 and p<0.01, respectively). CONCLUSIONS: The rosiglitazone-metformin combination significantly improved the long-term control of all insulin resistance-related parameters compared with the glimepiride metformin combination. However, both combinations were associated with a slight but statistically significant improvement in PAI-1 value, related to a similar reduction in insulin resistance. PMID- 15899725 TI - Effects of calcineurin inhibitors on sirolimus pharmacokinetics during staggered administration in renal transplant recipients. AB - STUDY OBJECTIVE: To compare the effects of different calcineurin inhibitors on sirolimus pharmacokinetics during long-term, staggered administration in kidney transplant recipients. Design. Randomized, open-label, parallel-group trial. SETTING: A medical center and one of its teaching hospitals in Taiwan. PATIENTS: Twenty-two de novo kidney transplant recipients. INTERVENTION: Patients received cyclosporine microemulsion or tacrolimus capsules twice/day in combination with once-daily sirolimus solution and corticosteroids. Sirolimus was administered 6 hours after the morning dose of cyclosporine or tacrolimus. After receiving a 6 mg loading dose of sirolimus, participants received sirolimus 2 mg/day for at least 7 days. Neither the cyclosporine nor the tacrolimus dosage was adjusted for at least 3 days before and during blood sampling for pharmacokinetic profiling. MEASUREMENTS AND MAIN RESULTS: One patient dropped out because of trimethoprim sulfamethoxazole-related hepatotoxicity. We observed no differences between the two patient groups in terms of their demographic data, renal and liver function, or dosage of sirolimus during the study. During multiple-dose administration, the area under the whole-blood concentration-time curve and the peak and trough concentrations of sirolimus in the cyclosporine group were, respectively, 1.46 (95% confidence interval [CI] 1.21-1.71), 1.42 (95% CI 1.08-1.76), and 1.42 (95% CI 1.09-1.76) times higher than those of the tacrolimus group, even though sirolimus was administered 6 hours after the other agents. CONCLUSION: Sirolimus pharmacokinetics may change significantly when calcineurin inhibitors are switched, even with staggered administration, which may not completely prevent a drug interaction between cyclosporine and sirolimus solution. PMID- 15899726 TI - Electrocardiographic effects of an Ephedra-Free, multicomponent weight-loss supplement in healthy volunteers. AB - STUDY OBJECTIVE: Metabolife 356, an ephedra-containing weight-loss product, substantially increases the corrected QT (QTc) interval. Metabolife Ephedra Free, a similar supplement, contains caffeine and extracts of green tea, garcinia cambogia, and yerba mate. Its electrocardiographic (ECG) effects are not known. Therefore, we sought to determine the effect of this supplement on the QTc interval. DESIGN: Randomized, double-blind, placebo-controlled, crossover study. SETTING: University of Connecticut, Storrs Campus. SUBJECTS: Twenty healthy volunteers. Intervention. A single capsule containing half the normal recommended dose of Metabolife Ephedra Free or matching placebo was administered in crossover fashion, with a 7-day washout period between treatments. MEASUREMENTS AND MAIN RESULTS: Baseline and three postdose ECG measurements were obtained, and QTc intervals were measured over a 5-hour study period. No significant differences in the QTc interval or other ECG variables were observed between the Metabolife Ephedra Free and placebo groups. CONCLUSION: At half the recommended single dose, Metabolife Ephedra Free does not affect the QTc interval or other ECG variables over 5 hours. Dose-response studies and studies of longer duration should be conducted. PMID- 15899727 TI - Persistence, switching, and discontinuation rates among patients receiving sertraline, paroxetine, and citalopram. AB - STUDY OBJECTIVE: To compare persistence, switching, and discontinuation rates among patients taking brand-name selective serotonin reuptake inhibitors (SSRIs). DESIGN: Retrospective cohort study. Data Source. Protocare Sciences managed care database. PATIENTS: A total of 14,933 patients with depression, posttraumatic stress disorder, or social anxiety disorder whose prescriptions for brand-name SSRIs were filled from January 1, 1999-June 30, 2002. MEASUREMENTS AND MAIN RESULTS: A total of 5598 patients received sertraline, 4131 citalopram, and 5204 paroxetine. Adherence status was categorized as persistence, switching (from one SSRI to another SSRI), and discontinuation. Persistence was determined based on the number of days' supply of the prescribed drug, with a minimum of 15 days to refill. Survival analyses using life-table survival curves and Cox proportional hazard models were conducted. Age, sex, and copayment were covariates in the Cox proportional models. Sensitivity analysis with a longer time to refill was performed to determine whether the results were sensitive to the algorithm for determining adherence. Compared with patients receiving sertraline and citalopram, those receiving paroxetine had lower rates of persistence (23.79% vs 25.96% for sertraline [p = 0.0093] and 26.56% for citalopram [p = 0.0022]) and higher rates of switching (3.55% vs 3.32% for sertraline [p = 0.5076] and 2.78% for citalopram [p = 0.0359]) and discontinuation (72.66% vs 70.72% for sertraline [p = 0.0258] and 70.66% for citalopram [p = 0.0334]). Survival curves showed that persistence rates with sertraline and citalopram were significantly greater than with paroxetine (p<0.05, log-rank and Wilcoxon tests). Age was an independent predictor of persistence; male sex and copayment were not. The comparisons across SSRIs were robust in the sensitivity analysis that varied the time to refill allowed. CONCLUSION: Differences in the persistence rates were noted among patients receiving three brand-name SSRIs, with patients receiving paroxetine having lower persistence than those receiving sertraline and citalopram. Observed differences in persistence across SSRIs were not sensitive to model specifications. Prescription of SSRIs that demonstrate better adherence would benefit both the patient and the health care system. PMID- 15899728 TI - Effect of clinical characteristics on neutropenia-related inpatient costs among newly diagnosed non-Hodgkin's lymphoma cases during first-course chemotherapy. AB - STUDY OBJECTIVE: To estimate the costs of hospitalization for neutropenia among chemotherapy-treated patients with newly diagnosed non-Hodgkin's lymphoma and to assess baseline patient factors associated with these costs. DESIGN: Retrospective cohort study. DATA SOURCE: Linked Surveillance, Epidemiology, and End Results Program-Healthcare Cost and Utilization Project databases for Iowa from 1993-1998. PATIENTS: Patients with newly diagnosed non-Hodgkin's lymphoma who received all inpatient care at Iowa hospitals during their first course of chemotherapy. MEASUREMENTS AND MAIN RESULTS: Neutropenia-related hospitalization costs were estimated from discharge abstracts found within the earliest of the following: 6 months after the diagnosis month, the date of bone marrow transplantation, or date of death. We performed univariate tests of differences in neutropenia-related hospitalization costs in all patients in the sample, as well as tests for neutropenia-related hospitalization costs, length-of-stay, and cost/inpatient day for patients with at least one hospitalization for neutropenia. We modeled total inpatient charges over the period for patients with at least one neutropenia-related hospitalization (multiple regression). A total of 1636 patients with non-Hodgkin's lymphoma had chemotherapy in Iowa and met inclusion criteria; of these, 316 had at least one hospitalization for neutropenia. The 316 patients had 418 stays. Patients with advanced stage (vs limited stage), previous anemia (vs no anemia), positive Charlson comorbidity score (vs score of 0), and diffuse large cell histology (vs follicular) had higher mean neutropenia-related hospitalization cost/patient with non-Hodgkin's lymphoma (p<0.05). Among those with neutropenia-related hospitalizations, a longer length of stay was associated with nonfollicular histologies, previous anemia, and positive Charlson score (p<0.05). CONCLUSION: When estimating expected payments for neutropenia-related hospitalization in patients with non Hodgkin's lymphoma, payers need to be aware of the distribution of clinical characteristics in these patients. PMID- 15899729 TI - Development and validation of a diabetes mellitus severity index: a risk adjustment tool for predicting health care resource use and costs. AB - STUDY OBJECTIVE: To develop and validate a diabetes mellitus-specific risk adjustment tool--the diabetes severity index (DSI)--to assist in predicting health care costs and resources within populations of patients with diabetes. DESIGN: Retrospective analysis of clinical and resource use for patients with a diagnosis of diabetes mellitus. Model estimation was conducted with half the sample, and validation analysis was conducted with the other half. SETTING: Southern Arizona Veterans Affairs Health Care System. PATIENTS: Seven hundred thirty-four patients with diabetes (710 men, 24 women; mean age 66 yrs). MEASUREMENTS AND MAIN RESULTS: Clinical measures of diabetes severity (known as the DSI) were used to predict three health care resource outcomes: risk of hospitalization, and total and ambulatory health care costs. Validity of the DSI was assessed by comparing the DSI with the revised chronic disease score (CDS). The DSI weights ranged from -471.5-3081.2 for total health care costs, from 304.3-1582.1 for outpatient costs, and -0.19-0.93 for risk of hospitalization. The DSI explained 6-8% of the variance in total and ambulatory costs and performed significantly (p<0.05) better than demographics alone, but was similar to the CDS. When the DSI was used with the CDS, up to 8% of variability in costs and use were explained. CONCLUSION: The DSI was developed to stratify veterans with diabetes according to diabetes severity and comorbidity. Since additional variance in medical care costs were explained by combining the DSI and CDS, the two indexes may explain different dimensions of a patient's severity of diabetes. However, the utility of such an index based on clinical measures to predict short term use may be limited due to low yet significant variability explained in the outcomes. PMID- 15899730 TI - Reduction in warfarin adverse events requiring patient hospitalization after implementation of a pharmacist-managed anticoagulation service. AB - STUDY OBJECTIVE: To compare adverse events related to anticoagulation in patients assigned to a pharmacist-managed anticoagulation service versus those receiving usual care. DESIGN: Retrospective cohort analysis. SETTING: Three hundred-bed community hospital. PATIENTS: Four hundred twenty patients referred for anticoagulation management. MEASUREMENTS AND MAIN RESULTS: Primary outcomes were the number of adverse events requiring patient hospitalization and the number of patients experiencing such events. Secondary outcomes were the median length of hospital stay/admission and the total number of hospital days. The total numbers of adverse events requiring hospitalization were three for the pharmacist-managed group and 14 for the usual care group (p=0.0153). The number of patients experiencing an adverse event requiring hospitalization was also lower for the pharmacist-managed group than for the usual care group (3 vs 10, p = 0.0962). The median length of hospital stay associated with each adverse event was not significantly different between the two groups; however, the total number of hospital days accrued was higher in the usual care group. CONCLUSION: At 6 months after discontinuation of the pharmacist-managed anticoagulation service, the frequency of adverse events increased significantly, resulting in both an increased number of hospitalizations and an increased number of hospital days accrued. This coordinated anticoagulation program using a pharmacist reduced warfarin-related complications. PMID- 15899731 TI - Experience with community-based amphotericin B infusion therapy. AB - STUDY OBJECTIVES: To identify the types and frequencies of adverse events associated with community-based amphotericin B infusion therapy. A second objective was to validate the effectiveness of a monitoring system, based on guidelines from the Infectious Diseases Society of America (IDSA). DESIGN: Retrospective medical record review. SETTING: Outpatient clinic at a tertiary care center. PATIENTS: One hundred five patients who received amphotericin B therapy from a home care provider between January 1997 and July 2002. MEASUREMENTS AND MAIN RESULTS: A total of 113 courses of amphotericin B formulations were administered: liposomal amphotericin B, 41 courses (36%), amphotericin B deoxycholate, 31 courses (27%), amphotericin B lipid complex, 31 courses (27%), and amphotericin B colloidal dispersion, 3 courses (3%); an additional 7 courses consisted of sequential therapy with two different formulations. Nephrotoxicity was associated with 46 (41%) courses, electrolyte abnormalities with 40 (35%) courses, venous access device complications with 12 (11%) courses, and infusion reactions with 13 (12%) courses. Nephrotoxicity occurred most frequently in adults aged 60 years or older, solid organ transplant recipients, and those receiving concomitant cyclosporine. Only two (12%) of 17 courses in children younger than 13 years were associated with nephrotoxicity. Thirteen of all 113 courses resulted in patients requiring hospital admission due to their adverse events. Monitoring of electrolyte, serum creatinine, and blood urea nitrogen levels 2 or 3 times/week was adequate for identifying these events. CONCLUSION: Significant rates of adverse events occurred in patients who received community-based amphotericin B infusion therapy. A monitoring system based on IDSA guidelines was effective in facilitating the detection and management of these adverse events. PMID- 15899732 TI - Use of highly active antiretroviral therapy in patients with renal insufficiency. AB - Antiretroviral agents, especially nucleoside reverse transcriptase inhibitors, require significant dosage adjustments in patients who have renal dysfunction and the human immunodeficiency virus (HIV). Some antiretroviral agents and fixed combination preparations are contraindicated in this population. In addition, many preferred antiretroviral regimens may be difficult to administer conveniently in patients with decreased creatinine clearance or in those receiving renal replacement therapies. Some highly active antiretroviral therapy regimens, however, can be used conveniently in patients with HIV and altered renal function. PMID- 15899733 TI - Cinacalcet: a new treatment for secondary hyperparathyroidism in patients receiving hemodialysis. AB - Cinacalcet is the first calcimimetic drug approved by the United States Food and Drug Administration for the treatment of secondary hyperparathyroidism in patients with chronic kidney disease. A literature search, performed by using PubMed and MEDLINE from January 1997-June 2004, identified articles concerning the efficacy and safety of cinacalcet in this patient population. Currently, Vitamin D and its analogs are considered first-line therapy for secondary hyperparathyroidism. However, use of these agents is often accompanied by an increase in serum calcium and phosphorus concentrations, a problem that often limits their use. Cinacalcet's mechanism of action decreases parathyroid hormone, calcium, and phosphorus levels, offering potential advantages over the other treatments for secondary hyperparathyroidism. Additional clinical trials are needed to evaluate the long-term safety and efficacy of the drug as a first-line agent. PMID- 15899734 TI - Gemifloxacin for the treatment of respiratory tract infections: in vitro susceptibility, pharmacokinetics and pharmacodynamics, clinical efficacy, and safety. AB - Gemifloxacin is a synthetic fluoroquinolone antimicrobial agent exhibiting potent activity against most gram-negative and gram-positive organisms, such as the important community-acquired respiratory pathogens Streptococcus pneumoniae (including multidrug-resistant S. pneumoniae), Haemophilus influenzae , and Moraxella catarrhalis . The agent's mechanism of action involves dual targeting of two essential bacterial enzymes: DNA gyrase and topoisomerase IV. Gemifloxacin was approved by the Food and Drug Administration in April 2003 for treatment of community-acquired pneumonia and acute bacterial exacerbation of chronic bronchitis. The drug has an oral bioavailability of approximately 71%. Approximately 20-35% of gemifloxacin is excreted unchanged in the urine after 24 hours. The elimination half-life of gemifloxacin is 6-8 hours in patients with normal renal function, supporting once-daily dosing. The 24-hour free-drug area under the plasma concentration-time curve:minimum inhibitory concentration ratio (fAUC(0-24):MIC) associated with efficacy, based on results from in vitro and animal models of infection, is approximately 30. With a mean fAUC(0-24) of approximately 3 microg*hour/ml (35% of total AUC(0-24) of 8.4) and a median S. pneumoniae MIC for 90% of tested strains of 0.03, a fAUC(0-24):MIC ratio of 100 would be expected after standard dosing (320 mg once/day). In clinical studies involving both hospitalized and outpatient populations, gemifloxacin has been highly effective in the treatment of community-acquired pneumonia and acute exacerbation of chronic bronchitis. Clinical success rates ranged from 93.9-95.9% in patients with community-acquired pneumonia and 96.1-97.5% in those with acute exacerbation of chronic bronchitis. Gemifloxacin is well tolerated; the frequency of adverse events with this agent is low. Most adverse events are mild-to moderate in severity, with diarrhea (< 4%), nausea and rash (< 3%), and headache (< 2%) most commonly reported. Drug interactions with gemifloxacin are not common, although absorption is greatly reduced when given with divalent and trivalent cation-containing compounds, such as antacids. Due to its potent activity against many common gram-positive and gram-negative respiratory pathogens, its proven clinical efficacy, and its favorable safety profile, gemifloxacin is a highly effective empiric treatment for community-acquired lower respiratory tract infections. PMID- 15899735 TI - Current issues in the development of a vaccine to prevent human immunodeficiency virus: insights from the society of infectious diseases pharmacists. AB - Infection with the human immunodeficiency virus (HIV) continues to affect millions of people worldwide. Preventive measures have done little to slow the transmission of the virus. Discovery of an effective vaccine to prevent HIV could have a tremendous impact on this global pandemic. However, the complex interactions between HIV and the host immune system have limited the progress in vaccine development. Traditional vaccination strategies have shown little promise. Currently, subunit vaccines, DNA vaccines, recombinant vector vaccines, and prime-boost strategies are being evaluated in clinical trials. Although many breakthroughs have been made in HIV vaccine research, only three candidate HIV vaccines have been studied in phase III clinical trials. The current strategies being investigated in the development of preventive HIV vaccines are reviewed. PMID- 15899736 TI - High-dose methotrexate in acute lymphoblastic leukemia: where is the evidence for its continued use? AB - High-dose intravenous methotrexate is an important component of many effective chemotherapeutic regimens for childhood acute lymphoblastic leukemia (ALL). Its use has a strong pharmacologic rationale: to overcome mechanisms of resistance of the malignant cells and to achieve cytotoxic concentrations in sanctuary sites for lymphoblasts. Although therapeutic progress in ALL during the past 4 decades has been closely associated with more widespread use of intravenous methotrexate and in progressively larger doses, little data exist to clearly support the use of high-dose intravenous methotrexate over a regimen of prolonged administration of low-dose methotrexate. The implied superiority of intravenous methotrexate mainly stems from studies that used identical leucovorin rescue with low-dose methotrexate or from studies of upfront window therapy in untreated children with ALL in which single standard doses of oral methotrexate were compared with high dose intravenous methotrexate with leucovorin rescue. The evidence favoring administration of intravenous methotrexate for children with ALL is critically reviewed. Despite its extensive use, high-dose intravenous methotrexate has not been proved conclusively to be more effective than less toxic, less labor intensive, and less costly methods of methotrexate administration. PMID- 15899737 TI - Dilated cardiomyopathy and acute liver injury associated with combined use of ephedra, gamma-hydroxybutyrate, and anabolic steroids. AB - Anabolic-androgenic steroids are synthetic derivatives of testosterone that some athletes have used to enhance muscle mass and improve their athletic performance. Ephedrine is a potent sympathomimetic agent that can lead to cardiomyopathy similar to that seen with catecholamine excess. Adverse cardiovascular events attributed to anabolic steroid and ephedra use, such as arrhythmias, myocardial infarction, cardiomyopathy, and sudden death, are rarely reported. Bodybuilders have used gamma-hydroxybutyrate, a potent secretagogue of growth hormone, to promote muscle development. Although dilated cardiomyopathy is a known complication of excess growth hormone levels, it has not been associated with use of gamma-hydroxybutyrate. A healthy 40-year-old man was admitted to our hospital for new-onset congestive heart failure and severe acute hepatitis that developed several months after he began using anabolic-androgenic steroids, ephedra, and gamma-hydroxybutyrate supplements. Analysis with an objective causality assessment scale revealed a probable adverse drug reaction between the patient's use of anabolic steroids, ephedra, and gamma-hydroxybutyrate and the development of his cardiomyopathy and acute liver injury. PMID- 15899738 TI - Lone atrial fibrillation associated with creatine monohydrate supplementation. AB - Atrial fibrillation in young patients without structural heart disease is rare. Therefore, when the arrhythmia is present in this population, reversible causes must be identified and resolved. Thyroid disorders, illicit drug or stimulant use, and acute alcohol intoxication are among these causes. We report the case of a 30-year-old Caucasian man who came to the emergency department in atrial fibrillation with rapid ventricular response. His medical history was unremarkable, except for minor fractures of the fingers and foot. Thyroid stimulating hormone, magnesium, and potassium levels were within normal limits, urine drug screen was negative, and alcohol use was denied. However, when the patient was questioned about use of herbal products and supplements, the use of creatine monohydrate was revealed. The patient was admitted to the hospital, anticoagulated with unfractionated heparin, and given intravenous diltiazem for rate control and intravenous amiodarone for rate and rhythm control. When discharged less than 24 hours later, he was receiving metoprolol and aspirin, with follow-up plans for echocardiography and nuclear imaging to assess perfusion. Exogenous creatine is used by athletes to theoretically improve exercise performance. Vegetarians may also take creatine to replace what they are not consuming from meat, fish, and other animal products. Previous anecdotal reports have linked creatine to the development of arrhythmia. Clinicians must be diligent when interviewing patients about their drug therapy histories and include questions about their use of herbal products and dietary supplements. In addition, it is important to report adverse effects associated with frequently consumed supplements and herbal products to the Food and Drug Administration and in the literature. PMID- 15899739 TI - Azacitidine-induced interstitial and alveolar fibrosis in a patient with myelodysplastic syndrome. AB - A 71-year-old Caucasian man diagnosed with myelodysplastic syndrome developed interstitial and alveolar fibrosis after receiving a 7-day course of azacitidine therapy. The patient's pulmonary function began to deteriorate immediately after the administration of his chemotherapy regimen. Other potential causes of pulmonary toxicity were ruled out such as viral, fungal, and bacterial pathogens, as well as other concomitant drugs. To our knowledge, this is the first case report documenting biopsy-proven interstitial and alveolar fibrosis associated with azacitidine. The frequency of this adverse drug reaction is unknown but may become more evident with increasing exposure of the population to azacitidine. PMID- 15899740 TI - Enoxaparin-associated retroperitoneal bleeding in two patients with renal insufficiency. AB - Two patients with chronic kidney disease experienced a major bleeding event, retroperitoneal hematoma, requiring a blood transfusion after the administration of enoxaparin. The first patient was a 61-year-old Caucasian woman with multiple comorbidities, including chronic kidney disease stage 4. She received subcutaneous enoxaparin 45 mg every 12 hours, along with antiplatelet agents. On the seventh day, she developed a large retroperitoneal hematoma and her hematocrit had decreased, requiring a transfusion of packed red blood cells. The second patient was a 74-year-old, obese Caucasian woman with multiple comorbidities, including chronic kidney disease stage 2-3 and atrial fibrillation. She was given enoxaparin 120 mg every 12 hours, along with warfarin and aspirin to prevent embolization. She developed a large retroperitoneal hematoma and died despite vigorous supportive care. Enoxaparin should be administered with great caution in patients with chronic kidney disease, especially if antiplatelet agents or other anticoagulants are administered concomitantly. PMID- 15899741 TI - Celecoxib-induced nephrotoxicity in a renal transplant recipient. AB - A 47-year-old renal transplant recipient came to the transplant clinic with a serum creatinine level that was elevated above her baseline value. She had been taking celecoxib for arthritic pain. She was told to discontinue the drug, and shortly after, her serum creatinine level returned to baseline. Several case reports describe nephrotoxicity with cyclooxygenase (COX)-2 inhibitors. However, only two of these reports involved renal transplant recipients, and in both, rofecoxib was the COX-2 inhibitor of concern. To our knowledge, this is the first case report of a renal transplant recipient who developed nephrotoxicity while taking celecoxib. The potential renal effects of COX-2 inhibitors have received little attention, even though nonsteroidal anti-inflammatory drugs are considered to carry the risk of nephrotoxicity in patients with comorbidities such as diabetes mellitus and hypertension. Further studies are necessary to determine the safety of COX-2 inhibitors in transplant recipients and other patient groups that may be at heightened risk of nephrotoxicity. PMID- 15899742 TI - Sedation guidelines: has the road map brought us to the destination? PMID- 15899743 TI - Daily interruption of continuous sedation. AB - Sedatives administered by continuous intravenous infusion are an essential component of the care of patients requiring mechanical ventilation. Delayed awakening from sedation has been associated with prolonged stays in the hospital and the intensive care unit (ICU). Incorporation of a daily sedation interruption policy into a medical ICU guideline has significantly reduced ICU stay and days of mechanical ventilation, and has significantly increased the likelihood of planned extubation compared with no intervention. Furthermore, opioid administration and ICU complications were significantly reduced in patients receiving daily sedation interruption. Lack of long-term deleterious psychiatric effects, such as evidence of posttraumatic stress disorder or recall of events, has been documented with daily sedation interruption during the ICU stay. Resistance to the routine implementation of a daily sedation interruption policy arises from concerns about the need for greater resources and the risk of rebound agitation. The benefits have been documented only in a single center to date. However, the benefit: risk ratio is positive and warrants incorporation of daily sedation interruption into the routine care of patients who are critically ill and require mechanical ventilation. PMID- 15899744 TI - Adverse events associated with sedatives, analgesics, and other drugs that provide patient comfort in the intensive care unit. AB - Since the 2002 publication of multidisciplinary clinical practice guidelines for intensive care unit (ICU) sedation and analgesia, additional information regarding adverse drug events has been reported. Our understanding of the risks associated with these sedative and analgesic agents promises to improve outcomes by helping clinicians identify and respond to therapeutic misadventures sooner. This review focuses on many issues, including the potentially fatal consequences associated with the propofol infusion syndrome, the evolving understanding of propylene glycol intoxication associated with parenteral lorazepam, new data involving high-dose and long-term dexmedetomidine therapy, haloperidol- and methadone-related prolongation of QTc intervals on the electrocardiogram, adverse events associated with atypical antipsychotics, and the potential for nonsteroidal antiinflammatory drugs to interfere with bone healing. PMID- 15899745 TI - Bispectral index monitoring in the intensive care unit provides more signal than noise. AB - The bispectral index (BIS) is processed electroencephalographic technology used in concert with clinical evaluations to objectively evaluate a patient's level of consciousness and probability of recall. Although the BIS has been extensively studied in the operating room setting, differences in patient populations, goals of treatment, and the environments themselves necessitate the development of BIS data specific to the intensive care unit. Data have evolved over the last several years, but for many reasons, the results and conclusions have varied. Yet within the data are important consistencies that help define the usefulness of BIS in patients who cannot be evaluated with subjective assessment tools such as the Sedation-Agitation Scale or the Richmond Agitation-Sedation Scale. Some of these patients cannot be evaluated with such tools because they lack motor responsiveness due to therapeutic paralysis or because they are receiving deep sedation. Bispectral index scores that are higher than expected in clinically sedated patients can often be traced to electromyographic activity or to the possibility of inadequate sedation and analgesia. The BIS must not be regarded as the sole indicator of level of consciousness, but should be used as part of an integrated approach to the evaluation of carefully selected patients with critical illness. PMID- 15899746 TI - Propofol: an immunomodulating agent. AB - Propofol (2,6-diisopropylphenol) is a potent intravenous hypnotic agent widely administered for induction and maintenance of anesthesia and for sedation in the intensive care unit. Propofol is insoluble in water and therefore is formulated in a lipid emulsion. In addition, a preservative (ethylenediaminetetraacetic acid [EDTA] or sodium metabisulfite) is added to retard bacterial growth. Propofol has antiinflammatory properties, decreasing production of proinflammatory cytokines, altering expression of nitric oxide, and inhibiting neutrophil function. Propofol also is a potent antioxidant. The added preservatives have biologic activity; EDTA has antiinflammatory properties, whereas metabisulfite may cause lipid peroxidation. The antiinflammatory and antioxidant properties of propofol may have beneficial effects in patients with sepsis and systemic inflammatory response syndrome. PMID- 15899747 TI - Sleep in the intensive care unit. AB - Adequate sleep, in terms of both quantity and quality, is an essential component of any overall health program. Sleep deprivation has serious deleterious effects on any subject, regardless of species. The most obvious and well-documented adverse effects of sleep deprivation are focused on cognitive function, cardiopulmonary function, and the immune system. Despite these deleterious effects of sleep deprivation, even on healthy subjects, intensive care patients are rarely permitted sufficient natural sleep. The causes of sleep disturbance in the intensive care unit involve factors related to the underlying clinical condition, those due to the various drugs required to provide treatment and comfort, and those related to the environment and processes of care in the intensive care unit. Only through a comprehensive approach to intensive care can an environment that promotes sleep and revitalization be constructed and maintained. PMID- 15899748 TI - An evaluation of a brief HIV/AIDS prevention intervention for college students using normative feedback and goal setting. AB - This study evaluated the ability of a 20-minute self-administered intervention to increase HIV/AIDS risk reduction among sexually active college students. The intervention presented normative data on the relatively low prevalence of HIV risk behaviors among college students for the purpose of conveying the idea that risk reduction was the prevailing social norm among their same age peers. The intervention also invited students to select specific risk reduction goals to be implemented over a 30-day follow-up period. Participants (N = 155) were assigned in alternating order to receive either the intervention or a control condition that entailed reading a general AIDS information pamphlet. Results were partially moderated by gender. Compared with controls, men in the intervention group reported significantly higher condom use, whereas women in the intervention group reported significantly fewer sexual partners. PMID- 15899749 TI - Cost-effectiveness of a behavioral intervention for seropositive youth. AB - An intervention for young people living with HIV (YPLH) was effective in reducing the number of partners of unknown serostatus and the number of unprotected sexual risk acts. In this article, we outline new methods to assess the cost effectiveness of this intervention. Over a period of 3 months, the intervention would avert an estimated 2.02 new infections per 1,000 YPLH. The cost of mounting the intervention was estimated at US 522 dollars/YPLH, with the cost effectiveness over a 1-year period being US 103,366 dollars/infection averted. Based on standardized estimates of the cost of treating HIV-positive persons and the adjusted quality of life years lost (10.23 for partners of a mean age of 29 years), the cost utility estimate shows that the treatment costs averted exceed the cost of the intervention. Both the methodology of calculating cost effectiveness and the cost utility of interventions are important for focusing policy makers, clinicians, community providers, and researchers on prevention for persons living with HIV. PMID- 15899750 TI - The prevalence, correlates, and costs of depression in people living with HIV/AIDS in Ontario: implications for service directions. AB - As new technologies extend the lives of people living with HIV/AIDS (PHA), the need increases for services that optimize their quality-of-life cost effectively. This study of PHAs (n = 297) in Ontario, Canada, examined the prevalence of depression, and its association with quality-of-life, coping strategies, social support, and use of health and social services. Results showed that depression was widespread (54.2%) and largely unrelated to demographic characteristics, but associated with diminished health status, health-related quality-of-life, and coping strategies. Depressed PHAs used significantly more crisis health care and related services, and community-based HIV/AIDS service organizations (ASOs). Findings suggest quality-of-life of PHAs may be improved by expanding the capacity of ASO workers to recognize and address depression, including helping depressed PHA access appropriate medication and sustain medication regimes. PMID- 15899751 TI - The employment perspectives study: identifying factors influencing the job seeking behavior of persons living with HIV/AIDS. AB - Given the increased quality and length of life of persons living with HIV/AIDS (PLWHA), this study explored factors influencing the job-seeking behaviors of PLWHA, using the theory of planned behavior. We conducted six focus groups with 54 persons living with HIV/AIDS who self-identified as African American, reported receiving HIV/AIDS services at an urban AIDS service organization in metropolitan Washington, DC, and reported not being employed. Most participants reported no employment history and were not receiving public assistance. Among the findings of this study, participants reported valuing employment, identified advantages and disadvantages associated with employment while managing their HIV/AIDS prognosis, identified individuals influencing their decision to become employed, and delineated facilitators and impediments to employment. The findings identified 15 beliefs that may be key for community-based organizations and agencies to address when providing job placement services for PLWHA. PMID- 15899752 TI - Risky sexual behavior among female entertainment workers in China: implications for HIV/STD prevention intervention. AB - Using qualitative information from in-depth interviews of 40 female entertainment workers (FEWs) in Shanghai, this article explores factors associated with unprotected sex and barriers to consistent condom use among FEWs. Results suggest that not all FEWs were at high risk, nor did they all engage in unprotected sex for the same reasons. Prevalence of unprotected sex varied by individual characteristics, across different settings where commercial sex took place, and by the FEWs' relationship to pimps or establishment owners. Factors contributing to unprotected sex included lack of HIV transmission knowledge, economic hardship, client refusal/coercion, and control by pimps/establishment owners. Incorrect beliefs also contributed to use of methods that offered no protection. Study participants endorsed the need for HIV/sexually transmitted disease prevention, but were suspicious about government programs. Prevention interventions need to target simultaneously clients of commercial sex and pimps/business owners and to promote a social environment supportive of 100% condom use in commercial sex. PMID- 15899753 TI - Changes in women's sexual risk behaviors after therapeutic community treatment. AB - This study examines sexual risk behaviors among 197 women mandated to substance abuse treatment in therapeutic communities. The women's risk behaviors after treatment are compared with their behaviors prior to treatment, and risk behaviors among those who completed treatment are compared with those who did not. The women had a high prevalence of sexual risk behaviors (i.e., multiple partners, unprotected sex, sex trade, not asking partners to use condoms, sex with injection drug users [IDUs]) and relationship violence and conflict before treatment and a significant decrease in unprotected sex, sex trade, sex with multiple partners, and relationship violence or conflict after treatment. Those who completed treatment were significantly less likely to participate in sex trade or have sex with IDUs and were more likely to have a new sexual partner and only one sexual partner after treatment as compared with those who did not complete treatment. The present study shows that therapeutic community treatment is associated with the reduction of sexual risk behaviors among female substance abusers mandated to treatment. PMID- 15899754 TI - Drug use and HIV risk practices of secondary and primary needle exchange users. AB - This study examines HIV risk practices associated with secondary needle exchange, obtaining needles from a needle exchange program (NEP) through others who attend in person. We analyzed data from NEP logs, a survey and HIV testing from 901 drug injectors who (a) always visited NEPs themselves to get needles (primary-only NEP users), (b) obtained at least some NEP needles by having others exchange for them (mixed/secondary NEP users), and (c) obtained no needles from an NEP. About 22% of 40,000 NEP visits involved secondary exchanges, and these accounted for over half of all needles exchanged. In multiple logistic regression analyses, primary only needle exchange was significantly associated with lower levels of receptive needle sharing, backloading, sharing other injection equipment and lending used needles, and positively associated with obtaining drug treatment. Mixed/secondary needle exchange was associated with less receptive needle sharing and a greater likelihood of drug treatment. Secondary exchange facilitated HIV risk reduction but the salutary effects of NEPs were attenuated in mixed/secondary exchangers. PMID- 15899755 TI - Psychodynamic psychotherapies: evidence-based practice and clinical wisdom. AB - This article is based on an exhaustive review of the psychotherapy outcomes literature, undertaken originally at the instigation of the UK Department of Health by Roth and Fonagy (Department of Health, 1995). We have recently updated this review (Fonagy, Target, Cottrell, Phillips, & Kurtz, 2002; Roth & Fonagy, 2004) and extended it to identify all studies of psychoanalytic psychotherapy. The usual methods for identifying studies were employed (Fonagy, Target, et al., 2002; Roth & Fonagy, in press). The key questions that should be asked of this literature given the current state of research in this area (also see Westen, Morrison, & Thompson-Brenner, 2004) are: Are there any disorders for which short term psychodynamic psychotherapy (STPP) can be considered evidence-based, Are there any disorders for which STPP is uniquely effective as either the only evidence-based treatment or as a treatment that is more effective than alternatives, and Is there any evidence base for long-term psychodynamic psychotherapy (LTPP) either in terms of achieving effects not normally associated with short-term treatment or addressing problems that have not been addressed by STPP? In this context, short-term therapy is conceived of as a treatment of around 20 sessions delivered usually once weekly. PMID- 15899756 TI - Rethinking adult attachment: a study of expert consensus. AB - Self-report questionnaires designed to assess attachment in adulthood potentially confound global valence of relationships (e.g., liking and disliking) with attachment. The authors surveyed an international panel of experts to develop items for a new measure of attachment that systematically distinguishes between attachment and non-attachment aspects of relationships. The survey yielded a large set of items on which there is consensus in five domains: secure attachment, dismissing attachment, preoccupied attachment, positive non attachment, and negative non-attachment. This report presents a content analysis of the core themes in each of these five domains and discusses implications of these results for conceptualizing and researching adult attachment. PMID- 15899757 TI - Symbolization and the good object. AB - Some patients struggle with the acceptance of, connection to, or identification with a good object who provides a containment-symbolization function. This is manifested as a standoff, a resistance, and a denial of the good object and, therefore, of the symbolization process that is part of healthy development. This is usually due to troubled family histories and a destructive projective identification process. The ego defensively blocks acknowledgment of any symbolizing capacity within the self and within the maternal object out of envy, fear of rejection, and dread of persecution. Destruction of symbolization and the experience of goodness within the self or in the object provides an artificial respite from the threat of annihilation, yet quickly brings on even greater levels of internal chaos and anxiety. Case material is used to illustrate these points. PMID- 15899758 TI - The two faces of the group therapist. AB - The group therapist wears two faces: spokesperson of the Establishment and spokesperson of significant truth. To run vital groups, these two roles of group leadership must stand in dialectic relationship to each other. The therapist introduces principles and practices that normalize group relations and provide a sense of cohesion, continuity, and regularity. However, to be constructive and promote significant truth, the group therapist must also be deconstructive, and encourage and support the group's questioning and challenging the very principles and practices that he or she asserts. A case example illustrates how the group leader is also two-faced in another sense of the term, since he or she may be false and insincere--or perceived as such--when being either the conservator or challenger of group process and culture. PMID- 15899759 TI - Long-term group psychotherapy for children with pervasive developmental disorders: evidence for group development. AB - This article highlights the clinical progress of three long-term psychotherapy groups for children with pervasive developmental disorders. Unique aspects of working with such children are addressed as well as their movement through the phases of group development. Clinical observations suggested that these groups did in fact pass through observable phases of cohesion, differentiation, and intimacy. Further, these children demonstrated noticeable social development both inside and outside the groups. They identified strongly with their groups, and after the settling-in period, there was remarkably little resistance to participation, especially given the children's tendencies for social avoidance. Vignettes are provided to illustrate both group developmental phases and noteworthy psychological experiences of the clinical process with children who have been diagnosed with pervasive developmental disorders. PMID- 15899760 TI - Joining a group's task: the discovery of hope and respect. AB - When engaged with others in tension-filled dialogue, the temptation is for us to treat the other disrespectfully by unconsciously choosing an exclusive subgroup in our minds to belong to, and assigning the other to a devalued group. Unconsciously asserted exclusive membership allows us to deny the ways in which, by disconnecting the other, we are also alienating parts of ourselves. Using three stories, the author explores the notion of unconscious group membership and raises the possibility of discovering a shared membership in the midst of conflict. Internalized shared group membership can constitute a mediating "third" to a dyad in apparent opposition. The shared internal context both contains and defines the pair within a task and set of values that can be used to transcend apparent conflict. PMID- 15899761 TI - At the core: how we steer our course. AB - "The force that through the green fuse drives the flower drives my green age..."- Dylan Thomas (1952). The beautiful evocative words of Dylan Thomas speak to the driving life force that pulses through all creation. So what, then, drives our organization, the American Group Psychotherapy Association (AGPA)? What underlying stream can we tap into to provide the energy, power, and vitality we need to go on being in a meaningful and productive fashion? And, what internal compass can we rely on to steer us in the right direction, to guide us in how we function as an organization? My purpose here is to consider how we formulate our objectives, determine our priorities, make decisions, and insure, as best we can, that we are pursuing that which is meaningful and essential, not veering drastically off course. To do this, I would like to focus briefly on the current organizational structure of AGPA, then examine the use of a strategic planning process, review our accomplishments, and lastly explore hopes and dreams for our future as the premier group psychotherapy organization in the United States. PMID- 15899762 TI - When the therapist says goodbye. AB - Therapist-initiated termination poses several unique problems for both the group and the therapist. This paper describes the termination process from the point of view of this therapist's experience in planning the termination of her group therapy practice. The reactions of other analysts to her decision to retire and the group processes and individual patient's reactions in this long-term psychoanalytic therapy group are described in detail. The author follows the group process and her own inner processes in the final six months of the group. The importance of the working through of the termination phase of therapy is supported by the experiences that were shared by the group and the therapist. PMID- 15899763 TI - Saying goodbye: exploring attachments as a therapist leaves a group of chronically ill persons. AB - This clinical report highlights some of the processes arising in a therapy group of persons with persistent and chronic psychiatric illnesses as they cope with the stress of their long-standing therapist's impending retirement. Members were initially disbelieving that the therapist would leave and were also concerned about their future care, particularly in terms of medications. Gradually they were more able to experience their feelings of loss and their genuine caring for the therapist and the group, using higher level defenses and increasing their ability to tolerate strong affects. They became increasingly able to demonstrate empathy, notably in their ability to consider the therapist's emotions. The clinical material also illustrates the therapist's personal involvement, countertransferences and expressions of concern. PMID- 15899769 TI - BBL CHROMagar Staph aureus is superior to mannitol salt for detection of Staphylococcus aureus in complex mixed infections. AB - We used 200 sputum specimens from patients with cystic fibrosis to evaluate BBL CHROMagar Staph aureus medium (CSA; BD Diagnostics, Spark, MD). Samples were inoculated to CSA, trypticase soy blood agar (BA), and Mannitol Salt (MS; BD Diagnostics). After 18 hours of incubation, CSA detected 39 (78%) of 50 Staphylococcus aureus (SA) samples; BA detected 30 (60%); and MS detected 29 (58%). Sensitivity after overnight incubation (at least 18 hours) was 82% for CSA, 72% for BA, and 71% for MS. At 2 days, CSA detected 48 (96%) of the SA. There were 2 false-positive results on CSA (99% specificity). There were 4 (8%) minor and no major or very major discrepancies between minimum inhibitory concentration (MIC) results for isolates grown on CSA and those grown on BA. Even at early reading times, CSA was superior to conventional media for detection of SA in these very complex cultures. MICs from all SA samples can be reported 24 hours sooner, and an additional BA subculture is not needed. PMID- 15899770 TI - A study of results generated using the Abbott LCx-GC assay fails to reveal a performance-based rationale for the 2002 level 1 recall. AB - To establish the effect of a quality control failure on the performance of the LCx-GC nucleic-acid amplification assay for Neisseria gonorrhoeae (Abbott Laboratories, Abbott Park, IL) in the field, we conducted a retrospective analysis comparing the clinical and analytic performance of the recalled lots with those not implicated in the recall. Our analysis revealed no statistically significant differences between recalled lots (n = 8,686 tests) and nonrecalled lots (n = 8,699 tests) with respect to multiple parameters of assay performance, including frequency distribution of patient results (P = .575), prevalence of indeterminate results (P = .245), mean positive control signals (P = .26), and within-run calibrator precision (P = .68). The LCx-GC system's lack of an electronic data storage and retrieval capability prevented assessment of the impact of the quality control failure on the clinical performance of recalled lots, such as the one described herein, from being conducted in real time. PMID- 15899771 TI - Bimodal cell populations are common in chronic lymphocytic leukemia but do not impact overall survival. AB - Flow cytometric histograms were evaluated for bimodal antigen expression on samples from 246 patients diagnosed with chronic lymphocytic leukemia (CLL) at University Hospitals of Cleveland, Cleveland, OH. Survival data were obtained, and the clinical significance of bimodality was evaluated using the Kaplan-Meier method and the log-rank test. Bimodal antigen expression was found in 107 cases (43.5%). CD38 and CD13 were the most common antigens to demonstrate bimodality at 14.5% and 12.9%, respectively, and CD20, CD11c, CD5, FMC-7, and surface immunoglobulin also were frequently bimodal. Bimodal antigen expression, the number of bimodal antigens, and bimodality of a specific antigen were not associated with decreased survival in patients with CLL, although bimodality for CD38 trended toward worse overall survival. Therefore, although bimodal antigen expression is common in CLL, the presence of bimodality does not seem to have significant prognostic importance PMID- 15899772 TI - Diagnostic usefulness of aberrant CD22 expression in differentiating neoplastic cells of B-Cell chronic lymphoproliferative disorders from admixed benign B cells in four-color multiparameter flow cytometry. AB - The diagnosis of B-cell chronic lymphoproliferative disorders is a great challenge when made in a background of polyclonal B cells. We studied the diagnostic usefulness of aberrant CD22 expression for differentiating neoplastic from benign B cells by 4-color flow cytometry. Of 56 cases of B-cell chronic lymphoproliferative disorders, we found that neoplastic cells showed aberrant CD22 expression in 39 (70%) of 56 cases, including chronic lymphocytic leukemia, mantle cell lymphoma, marginal zone lymphoma, hairy cell leukemia, and follicular lymphoma. In 4 cases, monoclonality was detected definitively only by evaluating the immunoglobulin light chain restriction in B cells with aberrant CD22 expression because numerous polyclonal B cells were present. Aberrant CD22 expression is a useful marker for detection of monoclonal B cells admixed with numerous benign polyclonal B cells. PMID- 15899773 TI - Studies of the site and distribution of CD34+ cells in idiopathic myelofibrosis. AB - The frequency and distribution of CD34+ cells in the bone marrow (BM) of patients with idiopathic myelofibrosis (IM) were determined using an immunohistochemical technique. The percentage and absolute number of circulating CD34+ cells were enumerated. Patients with IM exhibited a continuum of number of BM CD34+ cells ranging from 1 to 85 per 5 mm(2). The frequency of BM CD34+ cells was inversely related to the number of circulating CD34+ cells. The BM biopsy specimens obtained from 4 patients with IM who underwent allogeneic stem cell transplantation were examined sequentially. Quantitative measurement revealed that the reticulin fiber volume was progressively reduced after allogeneic stem cell transplantation. All 4 patients had normocellular marrow with normal numbers of BM CD34+ cells after transplantation. These findings suggest that the BM fibrosis and abnormal hematopoietic stem cell distribution in patients with IM is a consequence of the progeny of a malignant hematopoietic stem cell clone. PMID- 15899774 TI - Morphologic, cytogenetic, and molecular abnormalities in therapy-related acute promyelocytic leukemia. AB - We describe 17 cases of therapy-related acute promyelocytic leukemia (tAPL). Treatment for the initial neoplasms (mostly carcinomas and non-Hodgkin lymphomas) included radiation and chemotherapy in 11 patients, radiation in 3, and chemotherapy in 3. The interval between the initial neoplasm and tAPL ranged from 17 to 166 months (median, 40 months). Morphologically, all 13 cases with available bone marrow aspirate smears showed tAPL. Dyserythropoiesis or dysmegakaryopoiesis was identified in 11 cases. In 2 cases, too few nonneoplastic cells and, in all cases, too few maturing granulocytes were present to assess for dysplasia. Conventional cytogenetics or fluorescence in situ hybridization (FISH) showed the t(15;17)(q22;q21) in all cases; 6 as a sole abnormality, 9 with additional abnormalities, and 2 assessed only by FISH. Reverse transcription polymerase chain reaction (PCR) studies showed PML/RARa in 13 cases (8 short form, 5 long form). Mutations of the flt3 gene were detected by PCR in 5 (42%) of 12 cases. We conclude that dysplastic features, secondary cytogenetic abnormalities, and flt3 mutations are common in tAPL. PMID- 15899775 TI - Significance of stromal reaction patterns in invasive urothelial carcinoma. AB - We evaluated the types, frequency, and significance of stromal reaction patterns in urothelial carcinoma (UC) of the bladder in 60 transurethrally resected pT1 specimens (low-grade UC, 12; high-grade UC, 48). We observed 5 reaction patterns with 1 pattern in 37 cases (62%) and 2 or more patterns in the remainder. Dominant and secondary patterns, respectively, were as follows: stromal retraction, 30 (50%) and 4 (7%); edema, 18 (30%) and 1 (2%); inflammation, 8 (13%) and 14 (23%); fibroblastic proliferation, 3 (5%) and 5 (8%); fibrosis, 1 (2%) and 4 (7%). Progression occurred in 21 cases, including 9 (30%) of 30 with stromal retraction, 8 (45%) of 18 with edema, 2 (25%) of 8 with inflammation, 1 (33%) of 3 with fibroblastic proliferation, and 1 (100%) of 1 with fibrosis. Differences in progression rates and mean progression-free survival times were not statistically significant. We found that the most common stromal reaction in UC of the bladder is stromal retraction. Stromal reaction patterns seem to have some prognostic usefulness. Cases with stromal edema might benefit from closer follow-up. Awareness of the different types of stromal reactions also is useful for diagnosing invasion. PMID- 15899776 TI - The cytopathology of soft tissue mxyomas: ganglia, juxta-articular myxoid lesions, and intramuscular myxoma. AB - We studied the practicality of issuing a cytologic diagnosis of myxoma/juxta articular myxoid lesion/ganglion (MJG) by reviewing all fine-needle aspiration (FNA) biopsy specimens of soft tissue masses in our files with diagnoses of myxoma, myxoid cyst, myxoid lesion, ganglion, or ganglion cyst. The control group was soft tissue aspirates with abundant myxoid stroma. Of 39 cases with a cytologic diagnosis of soft tissue MJG, 15 had subsequent tissue biopsy or complete resection of the mass; 24 had clinicoradiologic follow-up. All cases except 1 (fat necrosis) were diagnosed correctly as benign myxoid lesions. We grouped MJG aspirates into 3 subtypes based on clinicoradiologic features: soft tissue ganglion/ganglion cyst (12 cases), juxta-articular myxoid lesion (16 cases), and intramuscular myxoma (11 cases). MJG aspirates showed few, subtle cytopathologic differences among subtypes. They characteristically had a viscous, gelatinous quality when expressed from the needle onto the glass slide. The typical smear contained a film of paucicellular, often finely granular, myxoid stroma, that contained few cells, usually macrophages or bland spindle cells. Control group aspirates always contained cellular components that allowed distinction from MJGs. The cytopathologic diagnosis of MJG lesions is accurate; FNA biopsy can be used to subtype MJGs into 3 categories when clinicoradiologic features are known. PMID- 15899777 TI - Cylindroma (dermal analog tumor) of the breast: a comparison with cylindroma of the skin and adenoid cystic carcinoma of the breast. AB - We compared 4 breast cylindromas with 50 dermal cylindromas and 8 adenoid cystic breast carcinomas. Except for a modest increase in the number of eccrine ducts and reactive Langerhans cells in dermal cylindromas, breast and dermal cylindromas showed identical histologic and immunohistochemical features. Both were characterized by epithelial islands containing central basaloid cells and peripheral myoepithelial cells surrounded by a thickened, continuous, periodic acid-Schiff-positive basement membrane that was immunoreactive for collagen IV. Clusters of sebaceous cells and a few eccrine ducts are described in breast cylindromas. Cytokeratin 7 labeled predominantly the central basaloid cells, and smooth muscle actin stained peripheral myoepithelial cells in breast and dermal cylindromas. Eccrine ducts were highlighted by epithelial membrane antigen and carcinoembryonic antigen. S-100 protein and CD1a showed a variable number of dendritic Langerhans cells. Cylindromas of the breast and skin did not express cytokeratin 20, gross cystic disease fluid protein 15, or estrogen or progesterone receptor. Breast cylindroma might be confused with the solid variant of adenoid cystic carcinoma, especially in needle core biopsy specimens, because they share nodular and trabecular patterns, basaloid cells, myoepithelial cells, eccrine ducts, and hyaline globules of basement membrane material. However, adenoid cystic carcinoma displays an infiltrative growth pattern, cytologic atypia, and mitotic figures and lacks the continuous, thickened basement membrane. PMID- 15899778 TI - Ki-67 immunoreactivity in the differential diagnosis of pulmonary neuroendocrine neoplasms in specimens with extensive crush artifact. AB - The aim of the present study was to evaluate the usefulness of immunohistochemical markers in the differential diagnosis of pulmonary neuroendocrine tumors with particular emphasis on the preservation of immunoreactivity in areas showing crush artifacts. Specimens from 9 carcinoid tumors (CTs) and 13 small cell carcinomas (SCCs) with crush artifact were stained with antibodies to Ki-67, chromogranin A, synaptophysin, and cytokeratin. The immunoreactivity was well preserved in the crushed areas. Ki-67 was expressed in the crushed areas of all SCCs. Reactivity was diffuse or at least present in 25% of the crushed areas. In contrast, the immunoreactive areas in CTs never exceeded 10%. Immunoreactivity for Ki-67, synaptophysin, chromogranin A, and cytokeratin is well preserved in tissue with crush artifacts and can be interpreted reliably. The diagnosis of SCC should be questioned if fewer than 25% of cells show reactivity for Ki-67. PMID- 15899779 TI - Expression of Fas ligand and bcl-2 in cervical carcinoma and their prognostic significance. AB - Apoptosis-related factors are known to influence survival with many malignant tumors. We performed immunohistochemical analysis of Fas ligand (FasL) and bcl-2 in invasive cervical cancer to determine the association with prognosis. In 125 patients with cervical cancer, 93 cases (74.4%) were positive for FasL, and 94 cases (75.2%) were positive for bcl-2. When 101 cases, clinical stages I through IV, were analyzed by univariate analysis, negative bcl-2 (P = .035) and combined positive FasL and negative bcl-2 (PFNB; P = .0025) were associated with significantly decreased disease-free survival. Positive FasL (P = .042), negative bcl-2 (P = .0004), and PFNB (P < .0001) were associated with a significantly worse prognosis in overall survival. In clinical stages II through IV, positive FasL (P = .04), negative bcl-2 (P = .002), and PFNB (P < .0001) had significant associations with shorter disease-free survival and positive FasL (P = .049), negative bcl-2 (P < .0001), and PFNB (P < .0001) with worse overall survival. PMID- 15899780 TI - Evaluation of initial and deeper sections of esophageal biopsy specimens for detection of intestinal metaplasia. AB - There are wide variations in the preparation of histologic sections from endoscopic esophageal biopsy specimens. We evaluated serial step sections from 261 esophageal or gastroesophageal junction biopsies at 4 levels to determine the first level at which goblet cell metaplasia (GCM) was detected. Deeper step sections of 152 paraffin blocks also were obtained to determine whether additional sections are useful in detecting GCM not seen in initial levels. GCM was identified in 95.3% of blocks in 3 levels. GCM was seen at level 4 in 12 blocks (4.7%). In the blocks that did not reveal intestinal metaplasia in the initial 4 levels, deeper sections disclosed GCM in only 1 (0.8%) of 120 blocks. However, deeper sections revealed initially undetected GCM in 4 of 32 blocks from patients with a history of documented Barrett esophagus. We conclude that 4 levels of step sections are adequate in routine processing of esophageal biopsy specimens for demonstration of GCM. Deeper sections may be obtained for patients with known Barrett esophagus to better evaluate for dysplasia or find additional foci of GCM. PMID- 15899781 TI - HER-2/neu and topoisomerase IIa gene amplification and protein expression in invasive breast carcinomas: chromogenic in situ hybridization and immunohistochemical analyses. AB - We studied HER-2/neu (HER-2) and topoisomerase IIa (topo2a) amplification (using chromogenic in situ hybridization) and overexpression (immunohistochemical analysis) in 113 invasive breast carcinomas. A gene copy number/chromosome 17 copy number ratio of 2.0 or higher indicated amplification. A topo2a/chromosome 17 ratio of less than 0.8 indicated gene deletion. HER-2 overexpression was scored according to standard HercepTest guidelines (DAKO, Carpinteria, CA). Overexpression of topo2a was identified when nuclear staining was found in more than 5% of tumor cells. Of 113 tumors, 104 were analyzed successfully for HER-2 and topo2a amplification. Of the 104, 64 showed HER-2 amplification; 25 of these (39%) also showed topo2a amplification. No amplification was found in 40 tumors. Deletion of topo2a was seen in 7 (11%) of 64 HER-2-amplified tumors and 2 (5%) of 40 nonamplified tumors. Of 25 tumors with topo2a amplification, 18 (72%) overexpressed topo2a. Only 3 (4%) of 79 tumors without topo2a amplification overexpressed topo2a. Amplification of topo2a is associated with HER-2 amplification but not vice versa. Amplification of topo2a resulted in protein overexpression in 72% of tumors, but topo2a overexpression rarely occurred without gene amplification. Identification of topo2a and HER-2 status might have therapeutic and prognostic implications. PMID- 15899782 TI - Evaluation of a modified reverse line blot assay for detection and typing of human papillomavirus. AB - Detection and typing of human papillomaviruses (HPVs) are requested by clinicians with growing frequency as part of the overall management of patients. A new method using consensus L1 amplification and a reverse line blot hybridization detection method has been described for broad spectrum HPV genotype identification. This method involves hybridization at 53 degrees C in a shaking water bath, which many laboratories might find cumbersome. We describe a modified hybridization step, using a dry-air incubator, that simplifies the detection protocol. Overall, comparable results were obtained by the modified, more easily performed protocol, indicating that laboratories might validate and use this modified method. PMID- 15899783 TI - Real-time quantitative PCR analysis of gene dosages reveals gene amplification in low-grade oligodendrogliomas. AB - Proto-oncogene amplification is an important alteration that is present in about 45% to 50% of high-grade human gliomas. We studied this mechanism in 8 genes (cyclin-dependent kinase-4 [CDK4], MDM2, MDM4, renin-angiotensin system-1, ELF3, GAC1, human epidermal growth factor receptor-2, and platelet-derived growth factor receptor-A gene) in a series of 40 oligodendrogliomas (World Health Organization (WHO) grade II, 21; WHO grade III, 13; and WHO grade II-III oligoastrocytomas, 6) using real-time quantitative polymerase chain reaction. Amplification of at least 1 of these genes was detected in 58% of samples (23/40). By histopathologic grade, 67% of grade II oligodendrogliomas (14/21), 46% of grade III anaplastic oligodendrogliomas (6/13), and 50% of mixed oligoastrocytomas (3/6) were positive for amplification of at least 1 gene. CDK4, MDM2, and GAC1 were the most frequently involved genes (12/40 [30%], 12/40 [30%], and 13/40 [33%], respectively). Our findings demonstrate gene amplification in low-grade samples indicating that it is an important alteration in the early steps of oligodendroglioma development and, therefore, might be considered a molecular mechanism leading to malignant progression toward anaplastic forms. PMID- 15899784 TI - Endogenous inhibitors of angiogenesis. AB - Angiogenesis, the formation of new blood vessels, is required for many pathologic processes, including invasive tumor growth as well as physiologic organ/tissue maintenance. Angiogenesis during development and adulthood is likely regulated by a balance between endogenous proangiogenic and antiangiogenic factors. It is speculated that tumor growth requires disruption of such balance; thus, the angiogenic switch must be turned "on" for cancer progression. If the angiogenic switch needs to be turned on to facilitate the tumor growth, the question remains as to what the physiologic status of this switch is in the adult human body; is it "off," with inhibitors outweighing the stimulators, or maintained at a fine "balance," keeping the proangiogenic properties of many factors at a delicate "activity" balance with endogenous inhibitors of angiogenesis. The physiologic status of this balance is important to understand as it might determine an individual's predisposition to turn the switch on during pathologic events dependent on angiogenesis. Conceivably, if the physiologic angiogenesis balance in human population exists somewhere between off and even balance, an individual's capacity and rate to turn the switch on might reflect their normal physiologic angiogenic status. In this regard, although extensive knowledge has been gained in our understanding of endogenous growth factors that stimulate angiogenesis, the activities associated with endogenous inhibitors are poorly understood. In this review, we will present an overview of the knowledge gained in studies related to the identification and characterization of 27 different endogenous inhibitors of angiogenesis. PMID- 15899785 TI - Lost in transcription: p21 repression, mechanisms, and consequences. AB - The cyclin-dependent kinase inhibitor p21WAF1/CIP1 is a major player in cell cycle control and it is mainly regulated at the transcriptional level. Whereas induction of p21 predominantly leads to cell cycle arrest, repression of p21 may have a variety of outcomes depending on the context. In this review, we concentrate on transcriptional repression of p21 by cellular and viral factors, and delve in detail into its possible biological implications and its role in cancer. It seems that the major mode of p21 transcriptional repression by negative regulators is the interference with positive transcription factors without direct binding to the p21 promoter. Specifically, the negative factors may either inhibit binding of positive regulators to the promoter or hinder their transcriptional activity. The ability of p21 to inhibit proliferation may contribute to its tumor suppressor function. Because of this, it is not surprising that a number of oncogenes repress p21 to promote cell growth and tumorigenesis. However, p21 is also an inhibitor of apoptosis and p21 repression may also have an anticancer effect. For example, c-Myc and chemical p21 inhibitors, which repress p21, sensitize tumor cells to apoptosis by anticancer drugs. Further identification of factors that repress p21 is likely to contribute to the better understanding of its role in cancer. PMID- 15899786 TI - Expression of kinase suppressor of Ras1 enhances cisplatin-induced extracellular signal-regulated kinase activation and cisplatin sensitivity. AB - Kinase suppressor of Ras1 (KSR1) interacts with several mitogen-activated protein (MAP) kinase pathway components, including Raf, MAP/extracellular signal regulated kinase (ERK) kinase (MEK), and ERK, and acts as a positive regulator of the Ras signaling cascade. Previous studies have shown that exposure of cells to the anticancer agent cisplatin (cis-diamminedichloroplatinum, CDDP) is associated with changes in multiple signal transduction pathways, including c-Jun-NH2 kinase, ERK, and p38 pathways. Moreover, ERK activation has been linked to changes in cell survival following CDDP treatment. In this report, we have examined the effects of KSR1 expression on the sensitivity of cells to CDDP induced apoptosis. Loss of KSR1 expression in mouse embryo fibroblasts (MEFs) derived from KSR1 knockout mice (KSR-/- MEF) is associated with decreased CDDP induced ERK activation and increased resistance to CDDP-induced apoptosis compared with wild-type MEFs (KSR+/+ MEF). Furthermore, transduction of KSR-/- MEFs and MCF-7 breast cancer cells with wild-type KSR1 resulted in enhanced ERK activation following CDDP exposure and increased sensitivity to CDDP. In addition, inhibition of ERK activation by exposing MEFs to the MEK1/2-specific inhibitors PD98059 and U0126 protected both KSR+/+ and KSR-/- MEFs cells from CDDP-induced apoptosis. These results indicate that KSR1-mediated regulation of ERK activity represents a novel determinant of CDDP sensitivity of cancer cells. PMID- 15899787 TI - Mesalazine improves replication fidelity in cultured colorectal cells. AB - Epidemiologic studies indicate that mesalazine has chemopreventive effects in inflammatory bowel disease-associated colorectal cancer. Most of our general understanding of chemoprevention in colorectal cancer is, however, derived from aspirin, which is structurally similar to mesalazine. Herein we determined the influence of aspirin and mesalazine on replication fidelity in cultured colorectal cells. Flow cytometry was used for quantitation of mutation rates at a (CA)13 microsatellite in HCT116 cells (mismatch repair deficient) and HCT116+chr3 cells (mismatch repair proficient) that had been stably transfected with pIREShyg2-EGFP/CA13, an enhanced green fluorescence protein-based plasmid, and cultured in the absence or presence of various concentrations of aspirin or mesalazine. Aspirin at doses above 1.25 mmol/L markedly reduced cell growth. Mesalazine doses up to 5.0 mmol/L had no such effect. The mutation rate in mismatch repair-deficient HCT116 cells was 6.8 x 10(-4) +/- 9.0 x 10(-5). In aspirin-treated cultures the mutation rate was 8.2 x 10(-4) +/- 1.3 x 10(-4) (121% of control). Instead, mesalazine lowered the mutation rate in a dose dependent fashion (5.5 x 10(-4) +/- 1.1 x 10(-4); 81% of control). The effects of mesalazine were most significant in the M1 fraction (P < 0.0001), which represents a mutant population immediate after the polymerase error and were confirmed in mismatch repair-proficient HCT116+chr3 cells. Our data indicate that mesalazine reduces frameshift mutations at a (CA)13 microsatellite in cultured colorectal cells independent of mismatch repair proficiency. This finding suggests that mesalazine improves replication fidelity, an effect that may be active in reducing mutations independent of its anti-inflammatory properties. PMID- 15899788 TI - CD4+CD25+ regulatory lymphocytes induce regression of intestinal tumors in ApcMin/+ mice. AB - Colorectal cancer in humans results from sequential genetic changes in intestinal epithelia commencing with inactivation of the APC tumor suppressor gene. Roles for host immunity in epithelial tumorigenesis are poorly understood. It has been previously shown that CD4+CD25+ lymphocytes inhibit colitis-associated epithelial tumors in Rag-deficient mice. Here we show that addition of CD4+CD25+ lymphocytes in ApcMin/+ mice reduces multiplicity of epithelial adenomas. Interleukin-10 was required in regulatory cells for therapeutic effect. Recipients of regulatory cells showed increased apoptosis and down-regulation of cyclooxygenase-2 within tumors coincident with tumor regression. These data suggest a role for regulatory lymphocytes in epithelial homeostasis in the ApcMin/+ mouse model of intestinal polyposis. Similarities with cancer of the breast, prostate, lung, and other sites raise the possibility of broader roles for regulatory lymphocytes in prevention and treatment of epithelial cancers in humans. PMID- 15899789 TI - Metastasizing melanoma formation caused by expression of activated N-RasQ61K on an INK4a-deficient background. AB - In human cutaneous malignant melanoma, a predominance of activated mutations in the N-ras gene has been documented. To obtain a mouse model most closely mimicking the human disease, a transgenic mouse line was generated by targeting expression of dominant-active human N-ras (N-RasQ61K) to the melanocyte lineage by tyrosinase regulatory sequences (Tyr::N-RasQ61K). Transgenic mice show hyperpigmented skin and develop cutaneous metastasizing melanoma. Consistent with the tumor suppressor function of the INK4a locus that encodes p16INK4A and p19(ARF), >90% of Tyr::N-RasQ61K INK4a-/- transgenic mice develop melanoma at 6 months. Primary melanoma tumors are melanotic, multifocal, microinvade the epidermis or epithelium of hair follicles, and disseminate as metastases to lymph nodes, lung, and liver. Primary melanoma can be transplanted s.c. in nude mice, and if injected i.v. into NOD/SCID mice colonize the lung. In addition, primary melanomas and metastases contain cells expressing the stem cell marker nestin suggesting a hierarchical structure of the tumors comprised of primitive nestin expressing precursors and differentiated cells. In conclusion, a novel mouse model with melanotic and metastasizing melanoma was obtained by recapitulating genetic lesions frequently found in human melanoma. PMID- 15899790 TI - SMARCB1/INI1 tumor suppressor gene is frequently inactivated in epithelioid sarcomas. AB - Epithelioid sarcoma is a rare soft tissue neoplasm of uncertain lineage that usually arises in the distal extremities of adults, presents a high rate of recurrences and metastases and frequently poses diagnostic dilemmas. The recently reported large-cell "proximal-type" variant is characterized by increased aggressiveness, deep location, preferential occurrence in proximal/axial regions of older patients, and rhabdoid features. Previous cytogenetic studies indicated that the most frequent alterations associated with this tumor entity affect chromosome 22. In this study, combined spectral karyotyping, fluorescence in situ hybridization, and array-based comparative genomic hybridization analyses of two proximal-type cases harboring a rearrangement involving 10q26 and 22q11 revealed that the 22q11 breakpoints were located in a 150-kb region containing the SMARCB1/INI1 gene, and that homozygous deletion of the gene was present in the tumor tissue. The SMARCB1/INI1 gene encodes for an invariant subunit of SWI/SNF chromatin remodeling complex and has been previously reported to act as a tumor suppressor gene frequently inactivated in infantile malignant rhabdoid tumors. We analyzed SMARCB1/INI1 gene status in nine additional epithelioid sarcoma cases (four proximal types and five conventional types) and altogether we identified deletions of SMARCB1/INI1 gene in 5 of 11 cases, all proximal types. We confirmed and further extended the number of cases with SMARCB1/INI1 inactivation to 6 of 11 cases, by real-time quantitative PCR analysis of mRNA expression and by SMARCB1/INI1 immunohistochemistry. Overall, these results point to SMARCB1/INI1 gene involvement in the genesis and/or progression of epithelioid sarcomas. Analysis of larger series of epithelioid sarcomas will be necessary to highlight putative clinically relevant features related to SMARCB1/INI1 inactivation. PMID- 15899791 TI - DNA ligase III as a candidate component of backup pathways of nonhomologous end joining. AB - Biochemical and genetic studies support the view that the majority of DNA double strand breaks induced in the genome of higher eukaryotes by ionizing radiation are removed by two pathways of nonhomologous end joining (NHEJ) termed D-NHEJ and B-NHEJ. Whereas D-NHEJ depends on the activities of the DNA-dependent protein kinase and DNA ligase IV/XRCC4, components of B-NHEJ have not been identified. Using extract fractionation, we show that the majority of DNA end joining activity in extracts of HeLa cells derives from DNA ligase III. DNA ligase III fractionates through two columns with the maximum in DNA end joining activity and its depletion from the extract causes loss of activity that can be recovered by the addition of purified enzyme. The same fractionation protocols provide evidence for an additional factor strongly enhancing DNA end joining and shifting the product spectrum from circles to multimers. An in vivo plasmid assay shows that DNA ligase IV-deficient mouse embryo fibroblasts retain significant DNA end joining activity that can be reduced by up to 80% by knocking down DNA ligase III using RNA interference. These in vivo and in vitro observations identify DNA ligase III as a candidate component for B-NHEJ and point to additional factors contributing to NHEJ efficiency. PMID- 15899792 TI - An expression-based site of origin diagnostic method designed for clinical application to cancer of unknown origin. AB - Gene expression profiling offers a promising new technique for the diagnosis and prognosis of cancer. We have applied this technology to build a clinically robust site of origin classifier with the ultimate aim of applying it to determine the origin of cancer of unknown primary (CUP). A single cDNA microarray platform was used to profile 229 primary and metastatic tumors representing 14 tumor types and multiple histologic subtypes. This data set was subsequently used for training and validation of a support vector machine (SVM) classifier, demonstrating 89% accuracy using a 13-class model. Further, we show the translation of a five-class classifier to a quantitative PCR-based platform. Selecting 79 optimal gene markers, we generated a quantitative-PCR low-density array, allowing the assay of both fresh-frozen and formalin-fixed paraffin-embedded (FFPE) tissue. Data generated using both quantitative PCR and microarray were subsequently used to train and validate a cross-platform SVM model with high prediction accuracy. Finally, we applied our SVM classifiers to 13 cases of CUP. We show that the microarray SVM classifier was capable of making high confidence predictions in 11 of 13 cases. These predictions were supported by comprehensive review of the patients' clinical histories. PMID- 15899793 TI - bHLH-zip transcription factor Spz1 mediates mitogen-activated protein kinase cell proliferation, transformation, and tumorigenesis. AB - BHLH-zip proteins usually play important regulatory roles in cell growth and differentiation. In this study, we show that Spz1, a bHLH-zip transcription factor, acts downstream of mitogen-activated protein kinase (MAPK, extracellular signal-regulated kinase 1/2) to up-regulate cell proliferation and tumorigenesis. In addition, through an interaction with proliferating cell nuclear antigen (PCNA) promoter, Spz1 induced cell proliferation concomitant with an increase in PCNA gene expression. Spz1-transfected cells formed colony foci on soft agar and developed fibrosarcoma tumors in nude mice. MAPK directly interacted and phosphorylated Spz1 protein, which increased PCNA transcription and cell tumorigenic activities. Reduction of endogenous Spz1 expression via RNA interference decreased cell proliferation in p19 embryonic carcinoma cells. High levels of Spz1 expression were detected in murine tumor cell lines and tumor samples of both human and Spz1 transgenic mice. Thus, Spz1 may act as a proto oncogene, participating in the MAPK signal pathway, and be a potential therapeutic target in the treatment of Ras-induced tumors. PMID- 15899794 TI - Gene expression profiling and genetic markers in glioblastoma survival. AB - Despite the strikingly grave prognosis for older patients with glioblastomas, significant variability in patient outcome is experienced. To explore the potential for developing improved prognostic capabilities based on the elucidation of potential biological relationships, we did analyses of genes commonly mutated, amplified, or deleted in glioblastomas and DNA microarray gene expression data from tumors of glioblastoma patients of age >50 for whom survival is known. No prognostic significance was associated with genetic changes in epidermal growth factor receptor (amplified in 17 of 41 patients), TP53 (mutated in 11 of 41 patients), p16INK4A (deleted in 15 of 33 patients), or phosphatase and tensin homologue (mutated in 15 of 41 patients). Statistical analysis of the gene expression data in connection with survival involved exploration of regression models on small subsets of genes, based on computational search over multiple regression models with cross-validation to assess predictive validity. The analysis generated a set of regression models that, when weighted and combined according to posterior probabilities implied by the statistical analysis, identify patterns in expression of a small subset of genes that are associated with survival and have value in assessing survival risks. The dominant genes across such multiple regression models involve three key genes-SPARC (Osteonectin), Doublecortex, and Semaphorin3B-which play key roles in cellular migration processes. Additional analysis, based on statistical graphical association models constructed using similar computational analysis methods, reveals other genes which support the view that multiple mediators of tumor invasion may be important prognostic factor in glioblastomas in older patients. PMID- 15899795 TI - A cell proliferation signature is a marker of extremely poor outcome in a subpopulation of breast cancer patients. AB - Breast cancer comprises a group of distinct subtypes that despite having similar histologic appearances, have very different metastatic potentials. Being able to identify the biological driving force, even for a subset of patients, is crucially important given the large population of women diagnosed with breast cancer. Here, we show that within a subset of patients characterized by relatively high estrogen receptor expression for their age, the occurrence of metastases is strongly predicted by a homogeneous gene expression pattern almost entirely consisting of cell cycle genes (5-year odds ratio of metastasis, 24.0; 95% confidence interval, 6.0-95.5). Overexpression of this set of genes is clearly associated with an extremely poor outcome, with the 10-year metastasis free probability being only 24% for the poor group, compared with 85% for the good group. In contrast, this gene expression pattern is much less correlated with the outcome in other patient subpopulations. The methods described here also illustrate the value of combining clinical variables, biological insight, and machine-learning to dissect biological complexity. Our work presented here may contribute a crucial step towards rational design of personalized treatment. PMID- 15899796 TI - E2F-dependent repression of topoisomerase II regulates heterochromatin formation and apoptosis in cells with melanoma-prone mutation. AB - RB pathway mutations, especially at the CDK4 and INK4A loci, are hallmarks of melanomagenesis. It is presently unclear what advantages these alterations confer during melanoma progression and how they influence melanoma therapy. Topoisomerase II inhibitors are widely used to treat human malignancies, including melanoma, although their variable success is attributable to a poor understanding of their mechanism of action. Using mouse and human cells harboring the melanoma-prone p16Ink4a-insensitive CDK4R24C mutation, we show here that topoisomerase II proteins are direct targets of E2F-mediated repression. Drug treated cells fail to load repressor E2Fs on topoisomerase II promoters leading to elevated topoisomerase II levels and an enhanced sensitivity of cells to apoptosis. This is associated with the increased formation of heterochromatin domains enriched in structural heterochromatin proteins, methylated histones H3/H4, and topoisomerase II. We refer to these preapoptotic heterochromatin domains as apoptosis-associated heterochromatic foci. We suggest that cellular apoptosis is preceded by an intermediary chromatin remodeling state that involves alterations of DNA topology by topoisomerase II enzymes and gene silencing via formation of heterochromatin. These observations provide novel insight into the mechanism of drug action that influence treatment outcome: drug sensitivity or drug resistance. PMID- 15899797 TI - YB-1 provokes breast cancer through the induction of chromosomal instability that emerges from mitotic failure and centrosome amplification. AB - YB-1 protein levels are elevated in most human breast cancers, and high YB-1 levels have been correlated with drug resistance and poor clinical outcome. YB-1 is a stress-responsive, cell cycle-regulated transcription factor with additional functions in RNA metabolism and translation. In this study, we show in a novel transgenic mouse model that human hemagglutinin-tagged YB-1 provokes remarkably diverse breast carcinomas through the induction of genetic instability that emerges from mitotic failure and centrosome amplification. The increase of centrosome numbers proceeds during breast cancer development and explanted tumor cell cultures show the phenotype of ongoing numerical chromosomal instability. These data illustrate a mechanism that might contribute to human breast cancer development. PMID- 15899798 TI - High-resolution genome-wide mapping of genetic alterations in human glial brain tumors. AB - High-resolution genome-wide mapping of exact boundaries of chromosomal alterations should facilitate the localization and identification of genes involved in gliomagenesis and may characterize genetic subgroups of glial brain tumors. We have done such mapping using cDNA microarray-based comparative genomic hybridization technology to profile copy number alterations across 42,000 mapped human cDNA clones, in a series of 54 gliomas of varying histogenesis and tumor grade. This gene-by-gene approach permitted the precise sizing of critical amplicons and deletions and the detection of multiple new genetic aberrations. It has also revealed recurrent patterns of occurrence of distinct chromosomal aberrations as well as their interrelationships and showed that gliomas can be clustered into distinct genetic subgroups. A subset of detected alterations was shown predominantly associated with either astrocytic or oligodendrocytic tumor phenotype. Finally, five novel minimally deleted regions were identified in a subset of tumors, containing putative candidate tumor suppressor genes (TOPORS, FANCG, RAD51, TP53BP1, and BIK) that could have a role in gliomagenesis. PMID- 15899799 TI - Detection of prostate cancer with a blood-based assay for early prostate cancer antigen. AB - Prostate-specific antigen lacks specificity for prostate cancer, so the identification and characterization of a unique blood-based marker for the disease would provide for a more accurate diagnosis, reducing both unnecessary biopsies and patient uncertainty. We previously identified a novel biomarker for prostate cancer, early prostate cancer antigen (EPCA). EPCA antibodies positively stained the negative biopsies of men who, as much as 5 years later, were diagnosed with prostate cancer. The goal of this study was to determine whether EPCA antibodies could be used in a clinically applicable plasma-based immunoassay to specifically detect prostate cancer. Using an EPCA-based ELISA, the protein was measured in the plasma of 46 individuals, including prostate cancer patients, healthy individuals, other cancer patients, spinal cord injury victims, and patients with prostatitis. With a predetermined cutoff value of 1.7 absorbance at 450 nm, only the prostate cancer population, as a whole, expressed plasma-EPCA levels above the cutoff. Statistical analysis showed a significant difference in EPCA levels between the prostate cancer population and each of the other groups, specifically the healthy donors (P < 0.0001), bladder cancer patients (P = 0.03), and spinal cord injury patients (P = 0.001). Sensitivity of the EPCA assay for prostate cancer patients was 92% whereas the overall specificity was 94%. Specificity for the healthy donors was 100%. Although larger trials are required, this initial study shows the potential of EPCA to serve as a highly specific blood-based marker for prostate cancer. EPCA, when coupled with prostate-specific antigen, may help reduce the number of both unnecessary biopsies and undetected prostate tumors. PMID- 15899800 TI - Association of DNA methylation of phosphoserine aminotransferase with response to endocrine therapy in patients with recurrent breast cancer. AB - To understand the biological basis of resistance to endocrine therapy is of utmost importance in patients with steroid hormone receptor-positive breast cancer. Not only will this allow us prediction of therapy success, it may also lead to novel therapies for patients resistant to current endocrine therapy. DNA methylation in the promoter regions of genes is a prominent epigenetic gene silencing mechanism that contributes to breast cancer biology. In the current study, we investigated whether promoter DNA methylation could be associated with resistance to endocrine therapy in patients with recurrent breast cancer. Using a microarray-based technology, the promoter DNA methylation status of 117 candidate genes was studied in a cohort of 200 steroid hormone receptor-positive tumors of patients who received the antiestrogen tamoxifen as first-line treatment for recurrent breast cancer. Of the genes analyzed, the promoter DNA methylation status of 10 genes was significantly associated with clinical outcome of tamoxifen therapy. The association of the promoter hypermethylation of the strongest marker, phosphoserine aminotransferase (PSAT1) with favorable clinical outcome was confirmed by an independent quantitative DNA methylation detection method. Furthermore, the extent of DNA methylation of PSAT1 was inversely associated with its expression at the mRNA level. Finally, also at the mRNA level, PSAT1 was a predictor of tamoxifen therapy response. Concluding, our work indicates that promoter hypermethylation and mRNA expression of PSAT1 are indicators of response to tamoxifen-based endocrine therapy in steroid hormone receptor-positive patients with recurrent breast cancer. PMID- 15899801 TI - Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) has nuclear localization signal-like sequences for nuclear import mediated by major vault protein. AB - Although phosphatase and tensin homologue deleted on chromosome 10 (PTEN) localization in the nucleus and cytoplasm is established, the mechanism is unknown. PTEN is a tumor suppressor phosphatase that causes cell cycle arrest and/or apoptosis. Nuclear-cytoplasmic compartmentalization may be a novel mechanism in regulating these events. PTEN does not contain a traditional nuclear localization sequence (NLS); however, we identified putative NLS-like sequences, which we analyzed by site-directed mutagenesis and localization studies in MCF-7 cells. Two double site mutations exhibited nuclear localization defects. Furthermore, unlike wild-type PTEN, double NLS mutant PTEN did not interact with major vault protein (MVP), a previously hypothesized nuclear-cytoplasmic transport protein. We conclude that these two NLS-like sequences are required for PTEN nuclear import that is mediated by MVP. Further, we show that this MVP mediated nuclear import is independent of PTEN phosphorylation and of the lipid and protein phosphatase activities of PTEN. PMID- 15899802 TI - BRCA2 regulates homologous recombination in response to DNA damage: implications for genome stability and carcinogenesis. AB - BRCA2 has been implicated in the maintenance of genome stability and RAD51 mediated homologous recombination repair of chromosomal double-strand breaks (DSBs), but its role in these processes is unclear. To gain more insight into its role in homologous recombination, we expressed wild-type BRCA2 in the well characterized BRCA2-deficient human cell line CAPAN-1 containing, as homologous recombination substrates, either direct or inverted repeats of two inactive marker genes. Whereas direct repeats monitor a mixture of RAD51-dependent and RAD51-independent homologous recombination events, inverted repeats distinguish between these events by reporting RAD51-dependent homologous recombination, gene conversion, and crossover events only. At either repeats, BRCA2 decreases the rate and frequency of spontaneous homologous recombination, but following chromosomal DSBs, BRCA2 increases the frequency of homologous recombination. At direct repeats, BRCA2 suppresses both spontaneous gene conversion and deletions, which can arise either from crossover or RAD51-independent sister chromatid replication slippage (SCRS), but following chromosomal DSBs, BRCA2 highly promotes gene conversion with little effect on deletions. At inverted repeats, spontaneous or DSB-induced crossover events were scarce and BRCA2 does not suppress their formation. From these results, we conclude that (i) BRCA2 regulates RAD51 recombination in response to the type of DNA damage and (ii) BRCA2 suppresses SCRS, suggesting a role for BRCA2 in sister chromatids cohesion and/or alignment. Loss of such control in response to estrogen-induced DNA damage after BRCA2 inactivation may be a key initial event triggering genome instability and carcinogenesis. PMID- 15899803 TI - Serum antibodies to huntingtin interacting protein-1: a new blood test for prostate cancer. AB - Huntingtin-interacting protein 1 (HIP1) is frequently overexpressed in prostate cancer. HIP1 is a clathrin-binding protein involved in growth factor receptor trafficking that transforms fibroblasts by prolonging the half-life of growth factor receptors. In addition to human cancers, HIP1 is also overexpressed in prostate tumors from the transgenic adenocarcinoma of the mouse prostate (TRAMP) mouse model. Here we provide evidence that HIP1 plays an important role in mouse tumor development, as tumor formation in the TRAMP mice was impaired in the Hip1null/null background. In addition, we report that autoantibodies to HIP1 developed in the sera of TRAMP mice with prostate cancer as well as in the sera from human prostate cancer patients. This led to the development of an anti-HIP1 serum test in humans that had a similar sensitivity and specificity to the anti alpha-methylacyl CoA racemase (AMACR) and prostate-specific antigen tests for prostate cancer and when combined with the anti-AMACR test yielded a specificity of 97%. These data suggest that HIP1 plays a functional role in tumorigenesis and that a positive HIP1 autoantibody test may be an important serum marker of prostate cancer. PMID- 15899804 TI - Gene expression profiling reveals cross-talk between melanoma and fibroblasts: implications for host-tumor interactions in metastasis. AB - Host-tumor interaction is considered critical in carcinogenesis, tumor invasion, and metastasis. To explore the reciprocal effects of host-tumor interaction, we developed a system to assess the gene expression patterns of A2058 human melanoma cells cocultured in fibrillar collagen with HS-68 primary human fibroblasts. The gene expression pattern of the cocultured A2058 cells was only modestly affected, whereas the HS-68 fibroblast gene expression pattern was significantly altered. Interleukin-11 and inhibitor of DNA-binding domain-1 gene expression in the cocultured A2058 cells was down-regulated, indicative of a proinflammatory response and resistance to apoptosis, respectively. The overall pattern of up regulated genes indicated triggering of the proinflammatory process. In addition, the melanoma growth and migration stimulatory chemokines CXCL1 and CXCL2 were significantly up-regulated in the cocultured fibroblasts. These results were corroborated by additional coculture experiments with the melanoma cell lines WM 164, BLM, and SK-Mel-28 and immunohistochemistry on invasive human melanoma sections. Taken together, these results indicate that tumor cells cause a proinflammatory and melanoma growth-promoting response in stromal fibroblasts. The role of inflammation in carcinogenesis, tumor promotion, invasion, and metastasis is viewed as being increasingly important and the results of these studies underscore this as well as identify certain key proteins that are expressed as a result of the complex interactive processes in the host-tumor microenvironment. PMID- 15899805 TI - The role of hypoxia-inducible factor-1 in three-dimensional tumor growth, apoptosis, and regulation by the insulin-signaling pathway. AB - The purpose of this study was to establish the effect of hypoxia-inducible factor 1 (HIF-1) directly on tumor growth, independently of angiogenesis. This was done by growing wild-type mouse hepatoma cells (HEPA-1) and their HIF-1-deficient counterpart C4 as multicellular tumor spheroids and quantifying differences in growth rates and proliferative and apoptotic indices. Insulin and insulin-like growth factor-I are key growth factors, also able to regulate hypoxia-responsive genes via HIF-1; thus, the effects of insulin on this model were also investigated. Two-dimensional growth was serum dependent and no difference was seen between wild-type HEPA-1 and C4 cell growth profiles, but major differences were seen in three-dimensional growth. HIF-1 supported spheroid growth under hypoxia as the HEPA-1 spheroids grew faster than the C4. Surprisingly, the HIF-1 deficient cells had a higher proliferation rate in three-dimensional growth (C4 mean S-phase index, 13.6%; HEPA-1 mean S-phase index, 9%; P = 0.009) that was associated with an inhibition of apoptosis. However, the apoptosis rate was much greater in these spheroids (C4 mean apoptotic index, 6.4; HEPA-1 mean apoptotic index, 0.78%; P = 0.0006). Addition of insulin increased proliferation and apoptosis in both HEPA-1 and C4 spheroids, demonstrating an HIF-1-independent effect of insulin signaling in three-dimensional growth. These results indicate that the enhancing effect of HIF-1 in three-dimensional tumor growth is a balance of both reduced proliferation and enhanced survival, the latter being proportionally greater. PMID- 15899806 TI - Regulation of tumor angiogenesis by fastatin, the fourth FAS1 domain of betaig h3, via alphavbeta3 integrin. AB - We previously reported that the FAS1 domains of betaig-h3 bear motifs that mediate endothelial cell adhesion and migration via interactions with alphavbeta3 integrin and regulate angiogenesis. In the present study, we show that the fourth FAS1 domain, designated fastatin, inhibits endothelial adhesion and migration, not only to betaig-h3, but also fibronectin and vitronectin, in a RGD-dependent manner. Fastatin and other FAS1 domains suppress endothelial cell tube formation and in vivo neovascularization in a Matrigel plug assay. The antiangiogenic activity of fastatin is associated with antitumor activity in mouse tumor models. Fastatin additionally induces apoptosis in several cells expressing alphavbeta3 integrin, including endothelial cells. Binding of fastatin to alphavbeta3 integrin inhibits phosphorylation of focal adhesion kinase, Raf, extracellular signal-regulated kinase, Akt, and mammalian target of rapamycin. Fastatin is thus the first endogenous angiogenesis regulator identified that inhibits both endothelial cell migration and growth by binding to alphavbeta3 integrin. Our data suggest that FAS1 domains from all possible forms of the four human FAS1 family proteins are potential endogenous regulators for pathologic angiogenesis. Moreover, FAS1 domains such as fastatin may be developed into drugs for blocking tumor angiogenesis. PMID- 15899807 TI - Human carcinoma cell growth and invasiveness is impaired by the propeptide of the ubiquitous proprotein convertase furin. AB - Furin, a potent proprotein convertase involved in activation of several cancer related substrates, is synthesized as an inactive zymogen, thus minimizing the occurrence of premature enzymatic activity that would lead to inappropriate protein activation or degradation. This natural inhibitory mechanism is based on the presence of an inactivating prosegment at the NH2 terminal of the zymogen. After initial autocatalytic cleavage, the prosegment remains tightly associated with the convertase until it reaches the trans-Golgi network where the dissociation of the prosegment and activation of furin occurs. We hypothesized that the inhibitory properties of the preprosegment of furin (ppFur) could be beneficial if ectopically expressed in tumor cells. Transfection of four human head and neck squamous cell carcinoma cell lines with the complete ppFur cDNA sequence (pIRES-EGFP-ppFur) or with the empty expression vector (pIRES-EGFP) was done. The inhibitory effect was evaluated using in vivo tumorigenicity, invasion, anchorage-independent growth in soft agar, and proliferation assays, as well as by investigating impairment of furin substrates processing. Following transfection of ppFur, a significant reduction in cell proliferation, tumorigenicity, and invasiveness was observed in vitro and in vivo. These biological changes are directly related to the inhibition of furin-mediated activation of crucial cancer-related substrates, such as membrane type 1 matrix metalloproteinase, transforming growth factor-beta, insulin-like growth factor-1 receptor, and vascular endothelial growth factor-C. PpFur expression in head and neck squamous cell carcinoma cell lines showed a mechanistic link between furin inhibition, decreased substrate processing, cell proliferation, and invasive ability. These findings suggest that furin inhibition is a feasible approach to ameliorate and even abolish the malignant phenotype of various malignancies. PMID- 15899808 TI - Expression of human endogenous retrovirus K in melanomas and melanoma cell lines. AB - The human endogenous retrovirus K family (HERV-K) comprises 30 to 50 closely related proviruses, most of which are defective. In contrast to all other human endogenous retroviruses, some HERV-K proviruses have maintained open reading frames for all viral proteins. In addition to the structural proteins Gag and Env and the reverse transcriptase, two regulatory proteins (Rec and Np9) have been described. Malignant melanoma has the highest mortality among skin cancers and is particularly aggressive. To study the expression of HERV-K, a set of seven primers was developed that allows discrimination between full-length and spliced mRNA and mRNA from deleted and undeleted proviruses. Expression of full-length mRNA from deleted and undeleted proviruses was detected in all human cells investigated. Expression of spliced env and rec was detected in a teratocarcinoma cell line, in 45% of the metastatic melanoma biopsies, and in 44% of the melanoma cell lines. In normal neonatal melanocytes, spliced rec was detected but not spliced env. Viral proteins were shown to be expressed in primary melanomas, metastases, and melanoma cell lines by immunohistochemistry, immunofluorescence, and Western blot analyses using specific antisera. For the first time, antibodies against HERV-K were found in melanoma patients. Melanomas are, in addition to teratocarcinomas and human breast cancer, the third tumor type with enhanced expression of HERV-K. PMID- 15899809 TI - Inhibitors of the arachidonic acid pathway and peroxisome proliferator-activated receptor ligands have superadditive effects on lung cancer growth inhibition. AB - Arachidonic acid (AA) metabolizing enzymes and peroxisome proliferator-activated receptors (PPARs) have been shown to regulate the growth of epithelial cells. We have previously reported that exposure to the 5-lipoxygenase activating protein directed inhibitor MK886 but not the cyclooxygenase inhibitor, indomethacin, reduced growth, increased apoptosis, and up-regulated PPARalpha and gamma expression in breast cancer cell lines. In the present study, we explore approaches to maximizing the proapoptotic effects of PPARgamma on lung cancer cell lines. Non-small-cell cancer cell line A549 revealed dose-dependent PPARgamma reporter activity after treatment with MK886. The addition of indomethacin in combination with MK886 further increases reporter activity. We also show increased growth inhibition and up-regulation of apoptosis after exposure to MK886 alone, or in combination with indomethacin and the PPAR ligand, 15-deoxy-Delta12,14-prostaglandin J2 compared with single drug exposures on the adenocarcinoma cell line A549 and small-cell cancer cell lines H345, N417, and H510. Real-time PCR analyses showed increased PPAR mRNA and retinoid X receptor (RXR)alpha mRNA expression after exposure to MK886 and indomethacin in a time dependent fashion. The results suggest that the principal proapoptotic effect of these drugs may be mediated through the known antiproliferative effects of the PPARgamma-RXR interaction. We therefore explored a three-drug approach to attempt to maximize this effect. The combination of low-dose MK886, ciglitazone, and 13 cis-retinoic acid interacted at least in a superadditive fashion to inhibit the growth of lung cancer cell lines A549 and H1299, suggesting that targeting PPARgamma and AA action is a promising approach to lung cancer growth with a favorable therapeutic index. PMID- 15899810 TI - Distinct structural domains within C19ORF5 support association with stabilized microtubules and mitochondrial aggregation and genome destruction. AB - C19ORF5 is a sequence homologue of microtubule-associated proteins MAP1A/MAP1B of unknown function, except for its association with mitochondria-associated proteins and the paclitaxel-like microtubule stabilizer and candidate tumor suppressor RASSF1A. Here, we show that when overexpressed in mammalian cells the recombinant 393-amino acid residue COOH terminus of C19ORF5 (C19ORF5C) exhibited four types of distribution patterns proportional to expression level. Although normally distributed throughout the cytosol without microtubular association, C19ORF5C specifically accumulated on stabilized microtubules in paclitaxel treated cells and interacted directly with paclitaxel-stabilized microtubules in vitro. The native 113-kDa full-length C19ORF5 and a shorter 56-kDa form similarly associated with stabilized microtubules in liver cells and stabilized microtubules from their lysates. As C19ORF5 accumulated, it appeared on mitochondria and progressively induced distinct perinuclear aggregates of mitochondria. C19ORF5 overlapped with cytochrome c-deficient mitochondria with reduced membrane potential. Mitochondrial aggregation resulted in gross degradation of DNA, a cell death-related process we refer to as mitochondrial aggregation and genome destruction (MAGD). Deletion mutagenesis revealed that the C19ORF5 hyperstabilized microtubule-binding domain resides in a highly basic sequence of <100 residues, whereas the MAGD activity resides further downstream in a distinct 25-residue sequence (F967-A991). Our results suggest that C19ORF5 mediates communication between the microtubular cytoskeleton and mitochondria in control of cell death and defective genome destruction through distinct bifunctional structural domains. The accumulation of C19ORF5 and resultant MAGD signaled by hyperstabilized microtubules may be involved in the tumor suppression activity of RASSF1A, a natural microtubule stabilizer and interaction partner with C19ORF5, and the taxoid drug family. PMID- 15899811 TI - Early detection of tumor response to chemotherapy by 3'-deoxy-3' [18F]fluorothymidine positron emission tomography: the effect of cisplatin on a fibrosarcoma tumor model in vivo. AB - We have assessed the potential of [18F]fluorothymidine positron emission tomography ([18F]FLT-PET) to measure early cytostasis and cytotoxicity induced by cisplatin treatment of radiation-induced fibrosarcoma 1 (RIF-1) tumor-bearing mice. Cisplatin-mediated arrest of tumor cell growth and induction of tumor shrinkage at 24 and 48 hours, respectively, were detectable by [18F]FLT-PET. At 24 and 48 hours, the normalized uptake at 60 minutes (tumor/liver radioactivity ratio at 60 minutes after radiotracer injection; NUV60) for [18F]FLT was 0.76 +/- 0.08 (P = 0.03) and 0.51 +/- 0.08 (P = 0.03), respectively, compared with controls (1.02 +/- 0.12). The decrease in [18F]FLT uptake at 24 hours was associated with a decrease in cell proliferation assessed immunohistochemically (a decrease in proliferating cell nuclear antigen labeling index, LI(PCNA), from 14.0 +/- 2.0% to 6.2 +/- 1.0%; P = 0.001), despite the lack of a change in tumor size. There were G1-S and G2-M phase arrests after cisplatin treatment, as determined by cell cycle analysis. For the quantitative measurement of tumor cell proliferation, [18F]FLT-PET was found to be superior to [18F]fluorodeoxyglucose PET (NUV60 versus LIPCNA: r = 0.89, P = 0.001 and r = 0.55, P = 0.06, respectively). At the biochemical level, we found that the changes in [18F]FLT and [18F]fluorodeoxyglucose uptake were due to changes in levels of thymidine kinase 1 protein, hexokinase, and ATP. This work supports the further development of [18F]FLT-PET as a generic pharmacodynamic readout for early quantitative imaging of drug-induced changes in cell proliferation in vivo. PMID- 15899812 TI - Role of hepatic cytochrome p450s in the pharmacokinetics and toxicity of cyclophosphamide: studies with the hepatic cytochrome p450 reductase null mouse. AB - Cyclophosphamide (CPA) is an anticancer prodrug that is dependent on cytochrome P450 (CYP) metabolism for its therapeutic effectiveness. In spite of the use of CPA in the clinic for over 50 years, little is known about the relationship between its toxicokinetics and therapeutic response. We have employed a powerful new model, the Hepatic Cytochrome P450 Reductase Null (HRN) mouse, which has almost no hepatic cytochrome P450 activity, to study the toxicokinetics of CPA and to establish in vivo the role of hepatic P450 metabolism in its pharmacokinetics. In HRN mice the in vitro metabolism and intrinsic clearance of CPA was over 6-fold lower than in wild-type animals. This change in CPA metabolism was also reflected in vivo, with a profound difference in the pharmacokinetics of both CPA and its metabolites. At a CPA dose of 100 mg/kg, the Cmax, plasma area under the curve (AUC) and half-life were increased by 2.6-, 6.2 , and 3.2-fold, respectively, in the HRN mice. Similar changes were also observed at a dose of 300 mg/kg. These data confirm that hepatic metabolism is the major route of CPA elimination and disposition. The primary metabolites of CPA, 4 hydroxycyclophosphamide (4-OH-CPA) and 3-dechloroethylcyclophosphamide, were still formed, but at altered rates in the HRN mice. At 100 mg/kg the t1/2 for 4 OH-CPA was increased 1.8-fold, the Cmax reduced 1.7-fold, and the AUC remained unchanged. This latter finding shows that P450-mediated oxidative metabolism is essential for the clearance of this compound. Toxicokinetic analysis of CPA induced myelosuppression and granulocytopenia showed that at high doses (> or =100 mg/kg) there was no difference in myelotoxicity between the wild-type and HRN mice. However, at lower doses (< or =70 mg/kg) a significant difference was observed, with little toxicity seen in HRN mice but at least a 45% reduction in the bone marrow granulocyte population in wild-type mice. Meta-analysis of the toxicity experiments showed the myelotoxicity of CPA was found to be closely correlated with the Cmax of 4-OH-CPA (r2= 0.80, P = 0.002). As the therapeutic effectiveness of CPA has been linked to the AUC for 4-OH-CPA, the finding that 4 OH-CPA Cmax may determine its level of myelotoxicity indicates that the therapeutic index could be altered by changing the method of CPA administration. Furthermore, monitoring 4-OH-CPA Cmax may identify individuals at most risk of CPA side effects. PMID- 15899813 TI - Functional epigenomics identifies genes frequently silenced in prostate cancer. AB - In many cases, silencing of gene expression by CpG methylation is causally involved in carcinogenesis. Furthermore, cancer-specific CpG methylation may serve as a tumor marker. In order to identify candidate genes for inactivation by CpG methylation in prostate cancer, the prostate cancer cell lines LNCaP, PC3, and Du-145 were treated with 5-aza-2' deoxycytidine and trichostatin A, which leads to reversion of epigenetic silencing. By microarray analysis of 18,400 individual transcripts, several hundred genes were found to be induced when compared with cells treated with trichostatin A. Fifty re-expressed genes were selected for further analysis based on their known function, which implied a possible involvement in tumor suppression. Twelve of these genes showed a significant degree of CpG methylation in their promoters. Six genes were silenced by CpG methylation in the majority of five analyzed prostate cancer cell lines, although they displayed robust mRNA expression in normal prostate epithelial cells obtained from four different donors. In primary prostate cancer samples derived from 41 patients, the frequencies of CpG methylation detected in the promoter regions of these genes were: GPX3, 93%; SFRP1, 83%; COX2, 78%; DKK3, 68%; GSTM1, 58%; and KIP2/p57, 56%. Ectopic expression of SFRP1 or DKK3 resulted in decreased proliferation. The expression of DKK3 was accompanied by attenuation of the mitogen-activated protein kinase pathway. The high frequency of CpG methylation detected in the promoters of the identified genes suggests a potential causal involvement in prostate cancer and may prove useful for diagnostic purposes. PMID- 15899814 TI - Transforming growth factor-beta suppresses nonmetastatic colon cancer through Smad4 and adaptor protein ELF at an early stage of tumorigenesis. AB - Although transforming growth factor-beta (TGF-beta) is both a suppressor and promoter of tumorigenesis, its contribution to early tumor suppression and staging remains largely unknown. In search of the mechanism of early tumor suppression, we identified the adaptor protein ELF, a beta-spectrin from stem/progenitor cells committed to foregut lineage. ELF activates and modulates Smad4 activation of TGF-beta to confer cell polarity, to maintain cell architecture, and to inhibit epithelial-to-mesenchymal transition. Analysis of development of colon cancer in (adult) elf+/-/Smad4+/-, elf+/-, Smad4+/-, and gut epithelial cells from elf-/- mutant mouse embryos pinpoints the defect to hyperplasia/adenoma transition. Further analysis of the role of ELF in human colorectal cancer confirms reduced expression of ELF in Dukes' B1 stage tissues (P < 0.05) and of Smad4 in advanced colon cancers (P < 0.05). This study indicates that by modulating Smad 4, ELF has a key role in TGF-beta signaling in the suppression of early colon cancer. PMID- 15899815 TI - Targeted expression of BRAFV600E in thyroid cells of transgenic mice results in papillary thyroid cancers that undergo dedifferentiation. AB - The BRAFT1799A mutation is the most common genetic alteration in papillary thyroid carcinomas (PTC). It is also found in a subset of papillary microcarcinomas, consistent with a role in tumor initiation. PTCs with BRAFT1799A are often invasive and present at a more advanced stage. BRAFT1799A is found with high prevalence in tall-cell variant PTCs and in poorly differentiated and undifferentiated carcinomas arising from PTCs. To explore the role of BRAFV600E in thyroid cancer pathogenesis, we targeted its expression to thyroid cells of transgenic FVB/N mice with a bovine thyroglobulin promoter. Two Tg-BRAFV600E lines (Tg-BRAF2 and Tg-BRAF3) were propagated for detailed analysis. Tg-BRAF2 and Tg-BRAF3 mice had increased thyroid-stimulating hormone levels (>7- and approximately 2-fold, respectively). This likely resulted from decreased expression of thyroid peroxidase, sodium iodine symporter, and thyroglobulin. All lines seemed to successfully compensate for thyroid dysfunction, as serum thyroxine/triiodothyronine and somatic growth were normal. Thyroid glands of transgenic mice were markedly enlarged by 5 weeks of age. In Tg-BRAF2 mice, PTCs were present at 12 and 22 weeks in 14 of 15 and 13 of 14 animals, respectively, with 83% exhibiting tall-cell features, 83% areas of invasion, and 48% foci of poorly differentiated carcinoma. Tg-BRAF3 mice also developed PTCs, albeit with lower prevalence (3 of 12 and 4 of 9 at 12 and 22 weeks, respectively). Tg-BRAF2 mice had a 30% decrease in survival at 5 months. In summary, thyroid-specific expression of BRAFV600E induces goiter and invasive PTC, which transitions to poorly differentiated carcinomas. This closely recapitulates the phenotype of BRAF-positive PTCs in humans and supports a key role for this oncogene in its pathogenesis. PMID- 15899816 TI - Real-time in vivo dual-color imaging of intracapillary cancer cell and nucleus deformation and migration. AB - The mechanism of cancer cell deformation and migration in narrow vessels is incompletely understood. In order to visualize the cytoplasmic and nuclear dynamics of cells migrating in capillaries, red fluorescent protein was expressed in the cytoplasm, and green fluorescent protein, linked to histone H2B, was expressed in the nucleus of cancer cells. Immediately after the cells were injected in the heart of nude mice, a skin flap on the abdomen was made. With a color CCD camera, we could observe highly elongated cancer cells and nuclei in capillaries in the skin flap in living mice. The migration velocities of the cancer cells in the capillaries were measured by capturing images of the dual color fluorescent cells over time. The cells and nuclei in the capillaries elongated to fit the width of these vessels. The average length of the major axis of the cancer cells in the capillaries increased to approximately four times their normal length. The nuclei increased their length 1.6 times in the capillaries. Cancer cells in capillaries over 8 microm in diameter could migrate up to 48.3 microm/hour. The data suggests that the minimum diameter of capillaries where cancer cells are able to migrate is approximately 8 microm. The use of the dual-color cancer cells differentially labeled in the cytoplasm and nucleus and associated fluorescent imaging provide a powerful tool to understand the mechanism of cancer cell migration and deformation in small vessels. PMID- 15899817 TI - HER2 overexpression increases sensitivity to gefitinib, an epidermal growth factor receptor tyrosine kinase inhibitor, through inhibition of HER2/HER3 heterodimer formation in lung cancer cells. AB - Gefitinib (Iressa), an epidermal growth factor receptor targeting drug, has been clinically useful for the treatment of patients with non-small cell lung cancer (NSCLC). Gefitinib is currently being applied in clinical studies as either a monotherapy, or as part of a combination therapy against prostate, head and neck, gastric, breast, and colorectal tumors. However, success rates vary between different tumor types, and thus it is important to understand which molecular target(s) are responsible for limiting the therapeutic efficacy of the drug. In this study, we ask whether expression of HER2 affects sensitivity to gefitinib in human lung cancer cells. We established two clones, LK2/HER2-32 and LK2/HER2-57, by transfecting HER2 cDNA into LK2, a NSCLC line with a low expression level of HER2. We observed no mutations in exons 18, 19, and 21 of EGFR gene in LK2, LK2/mock- and two HER2-trasfectants when we observed in-frame deletion mutations (E746-A750) adjacent to K745 in a gefitinib-sensitive NSCLC cell line, PC9. These LK2/HER2-32 and LK2/HER2-57 were much more sensitive to the cytotoxic effects of gefitinib than the parental LK2 lines. Treatment with 0.5 to 1 micromol/L gefitinib specifically blocked Akt activation in both HER2-transfectant lines, but not in the parental LK2 cells. Extracellular signal-regulated kinase-1/2 activation, however, was not blocked by gefitinib up to 10 micromol/L in either the parent or transfectant lines. Gefitinib was also shown to induce cell cycle arrest in the G1-S phase, and an accompanying increase of p27Kip1 was observed. LK2/HER2 transfectants showed constitutive formation of HER2/HER3 heterodimer, which were seen to associate with a regulatory subunit of phosphoinositide-3 kinase, p85alpha, when active. Treatment of LK2/HER2 cells with gefitinib markedly decreased the formation of HER2/HER3 heterodimers, HER3 basal phosphorylation, and the association of p85alpha with HER3. This study is the first to show that under basal growth conditions, HER2 sensitizes low-EGFR NSCLC cell lines to growth inhibition by gefitinib. PMID- 15899818 TI - Matrix metalloproteinases 9 and 10 inhibit protein kinase C-potentiated, p53 mediated apoptosis. AB - p53, a major sensor of DNA damage, is a transcription factor that, depending on its phosphorylation status, regulates the cell cycle, DNA repair, or apoptosis. The protein kinase C (PKC) family of isozymes is also implicated in cell cycle and programmed cell death (PCD) control and has recently been shown to influence p53 function. Using three human colon adenocarcinoma cell lines SW480, EB-1, and HCT116 that either lack p53 function and were engineered to express inducible wild-type p53 (wt p53), or that constitutively express wt p53, we show that phorbol ester-mediated PKC activation potentiates p53-induced PCD. Despite the effectiveness of PKC/p53 synergy in inducing SW480 tumor cell death, however, a fraction of the cells invariably survive. To address the putative mechanisms that underlie resistance to PKC/p53-induced cell death, we generated a phorbol 12 myristate 13-acetate/p53-resistant SW480 subline and compared the gene expression profile of resistant and parental cells by DNA microarray analysis. The results of these experiments show that PKC/p53-resistant cells express a higher level of several matrix metalloproteinases (MMP), including MMP-9, MMP-10, and MMP-12, and corresponding real-time PCR assays indicate that p53 is a negative regulator of MMP-9 gene expression. Using MMP inhibitors and MMP-specific small interfering RNA, we show that MMP function confers protection from PKC/p53-induced apoptosis and identify the protective MMPs as MMP-9 and MMP-10. Taken together, these observations provide evidence that MMPs are implicated in tumor cell resistance to the synergistic proapoptotic effect of PKC and p53. PMID- 15899819 TI - Nuclear factor-kappaB induced by doxorubicin is deficient in phosphorylation and acetylation and represses nuclear factor-kappaB-dependent transcription in cancer cells. AB - The primary goal of chemotherapy is to cause cancer cell death. However, a side effect of many commonly used chemotherapeutic drugs is the activation of nuclear factor-kappaB (NF-kappaB), a potent inducer of antiapoptotic genes, which may blunt the therapeutic efficacy of these compounds. We have assessed the effect of doxorubicin, an anthracycline in widespread clinical use, on NF-kappaB activation and expression of antiapoptotic genes in breast cancer cells. We show that doxorubicin treatment activates NF-kappaB signaling and produces NF-kappaB complexes that are competent for NF-kappaB binding in vitro. Surprisingly, these NF-kappaB complexes suppress, rather than activate, constitutive- and cytokine induced NF-kappaB-dependent transcription. We show that doxorubicin treatment produces RelA, which is deficient in phosphorylation and acetylation and which blocks NF-kappaB signaling in a histone deacetylase-independent manner, and we show that NF-kappaB activated by doxorubicin does not remain stably bound to kappaB elements in vivo. Together these data show that NF-kappaB signaling induced by doxorubicin reduces expression of NF-kappaB-dependent genes in cancer cells. PMID- 15899820 TI - Stimulation of mitochondrial activity by p43 overexpression induces human dermal fibroblast transformation. AB - Mitochondrial dysfunctions are frequently reported in cancer cells, but their direct involvement in tumorigenesis remains unclear. To understand this relation, we stimulated mitochondrial activity by overexpression of the mitochondrial triiodothyronine receptor (p43) in human dermal fibroblasts. In all clones, this stimulation induced morphologic changes and cell fusion in myotube-like structures associated with the expression of several muscle-specific genes (Myf5, desmin, connectin, myosin, AchRalpha). In addition, these clones displayed all the in vivo and in vitro features of cell transformation. This phenotype was related to an increase in c-Jun and c-Fos expression and extinction of tumor suppressor gene expression (p53, p21WAF1, Rb3). Lastly, reactive oxygen species (ROS) production was increased in positive correlation to the stimulation of mitochondrial activity. The direct involvement of mitochondrial activity in this cell behavior was studied by adding chloramphenicol, an inhibitor of mitochondrial protein synthesis, to the culture medium. This inhibition resulted in partial restoration of the normal phenotype, with the loss of the ability to fuse, a strong decrease in muscle-specific gene expression, and potent inhibition of the transformed phenotype. However, expression of tumor suppressor genes was not restored. Similar results were obtained by using N-acetylcysteine, an inhibitor of ROS production. These data indicate that stimulation of mitochondrial activity in human dermal fibroblasts induces cell transformation through events involving ROS production. PMID- 15899821 TI - alpha-Melanocortin and endothelin-1 activate antiapoptotic pathways and reduce DNA damage in human melanocytes. AB - UV radiation is an important etiologic factor for skin cancer, including melanoma. Constitutive pigmentation and the ability to tan are considered the main photoprotective mechanism against sun-induced carcinogenesis. Pigmentation in the skin is conferred by epidermal melanocytes that synthesize and transfer melanin to keratinocytes. Therefore, insuring the survival and genomic stability of epidermal melanocytes is critical for inhibiting photocarcinogenesis, particularly melanoma, the most deadly form of skin cancer. The paracrine factors alpha-melanocortin and endothelin-1 are critical for the melanogenic response of cultured human melanocytes to UV radiation. We report that alpha-melanocortin and endothelin-1 rescued human melanocytes from UV radiation-induced apoptosis and reduced DNA photoproducts and oxidative stress. The survival effects of alpha melanocortin and endothelin-1 were mediated by activation of the melanocortin 1 and endothelin receptors, respectively. Treatment of melanocytes with alpha melanocortin and/or endothelin-1 before exposure to UV radiation activated the inositol triphosphate kinase-Akt pathway and increased the phosphorylation and expression of the microphthalmia-related transcription factor. Treatment with alpha-melanocortin and/or endothelin-1 enhanced the repair of cyclobutane pyrimidine dimers and reduced the levels of hydrogen peroxide induced by UV radiation. These effects are expected to reduce genomic instability and mutagenesis. PMID- 15899822 TI - Synergistic antitumor response of interleukin 2 with melphalan in isolated limb perfusion in soft tissue sarcoma-bearing rats. AB - The cytokine interleukin 2 (IL-2) is a mediator of immune cell activation with some antitumor activity, mainly in renal cell cancer and melanoma. We have previously shown that tumor necrosis factor (TNF)-alpha has strong synergistic antitumor activity in combination with chemotherapeutics in the isolated limb perfusion (ILP) setting based on a TNF-mediated enhanced tumor-selective uptake of the chemotherapeutic drug followed by a selective destruction of the tumor vasculature. IL-2 can cause vascular leakage and edema and for this reason we examined the antitumor activity of a combined treatment with IL-2 and melphalan in our well-established ILP in soft tissue sarcoma-bearing rats (BN175). ILP with either IL-2 or melphalan alone has no antitumor effect, but the combination of IL 2 and melphalan resulted in a strong synergistic tumor response, without any local or systemic toxicity. IL-2 enhanced significantly melphalan uptake in tumor tissue. No signs of significant vascular damage were detected to account for this observation, although the tumor sections of the IL-2- and IL-2 plus melphalan treated animals revealed scattered extravasation of erythrocytes compared with the untreated animals. Clear differences were seen in the localization of ED-1 cells, with an even distribution in the sham, IL-2 and melphalan treatments, whereas in the IL-2 plus melphalan-treated tumors clustered ED-1 cells were found. Additionally, increased levels of TNF mRNA were found in tumors treated with IL-2 and IL-2 plus melphalan. These observations indicate a potentially important role for macrophages in the IL-2-based perfusion. The results in our study indicate that the novel combination of IL-2 and melphalan in ILP has synergistic antitumor activity and may be an alternative for ILP with TNF and melphalan. PMID- 15899823 TI - Versatile prostate cancer treatment with inducible caspase and interleukin-12. AB - To establish optimized conditions for immunity against prostate cancer, we compared the efficacy of multiple approaches in autochthonous and s.c. transgenic adenocarcinoma of the mouse prostate (TRAMP)-based models. Mice immunized with interleukin (IL)-12-containing apoptotic, but not necrotic TRAMP-C2 cell-based, vaccines were resistant to TRAMP-C2 tumor challenge and re-challenge, independently of the route of vaccination (s.c. or i.p.). Administration of gamma irradiated TRAMP-C2 cells preinfected with adenovirus containing both B7-1 and IL 12 genes, unlike adenovirus containing B7-1 alone, considerably protected C57BL/6 mice from TRAMP-C2 tumor growth and extended the life span of TRAMP mice. Vaccines that included dendritic cells, instead of IL-12, were equally efficient. Whereas injections of ligand-inducible caspase-1- and IL-12-containing adenoviruses cured small s.c. TRAMP-C2 tumors, nanopump-regulated delivery of viruses led to elimination of much larger tumors. The antitumor immune responses involved CD4+-, CD8+-, and natural killer cells and were strengthened by increasing the number of vaccinations. Intraprostatic administration of inducible caspase-1- and IL-12-containing adenoviruses resulted in local cell death and improved survival of adenocarcinoma-bearing TRAMP mice. Thus, tumor cell apoptosis induced by caspase in situ and accompanied by IL-12 is efficient against prostate cancer in a preclinical model. PMID- 15899824 TI - ABCB5-mediated doxorubicin transport and chemoresistance in human malignant melanoma. AB - Enhanced drug efflux mediated by ABCB1 P-glycoprotein and related ATP-binding cassette transporters is one of several mechanisms of multidrug resistance thought to impair chemotherapeutic success in human cancers. In malignant melanoma, its potential contribution to chemoresistance is uncertain. Here, we show that ABCB5, which functions as a determinant of membrane potential and regulator of cell fusion in physiologic skin progenitor cells, is expressed in clinical malignant melanoma tumors and preferentially marks a subset of hyperpolarized, CD133+ stem cell phenotype-expressing tumor cells in malignant melanoma cultures and clinical melanomas. We found that ABCB5 blockade significantly reversed resistance of G3361 melanoma cells to doxorubicin, an agent to which clinical melanomas have been found refractory, resulting in a 43% reduction in the LD50 from 4 to 2.3 micromol/L doxorubicin (P < 0.05). Our results identified ABCB5-mediated doxorubicin efflux transport as the underlying mechanism of resistance, because ABCB5 blockade significantly enhanced intracellular drug accumulation. Consistent with this novel ABCB5 function and mechanism in doxorubicin resistance, gene expression levels of the transporter across a panel of human cancer cell lines used by the National Cancer Institute for drug screening correlated significantly with tumor resistance to doxorubicin (r = 0.44; P = 0.016). Our results identify ABCB5 as a novel drug transporter and chemoresistance mediator in human malignant melanoma. Moreover, our findings show that ABCB5 is a novel molecular marker for a distinct subset of chemoresistant, stem cell phenotype-expressing tumor cells among melanoma bulk populations and indicate that these chemoresistant cells can be specifically targeted via ABCB5 to enhance cytotoxic efficacy. PMID- 15899825 TI - Treatment of chemotherapy-resistant human ovarian cancer xenografts in C.B 17/SCID mice by intraperitoneal administration of Clostridium perfringens enterotoxin. AB - Ovarian cancer remains the most lethal gynecologic malignancy in the United States. Although many patients with advanced-stage disease initially respond to standard combinations of surgical and cytotoxic therapy, nearly 90% develop recurrence and inevitably die from the development of chemotherapy-resistant disease. The discovery of novel and effective therapy against chemotherapy resistant/recurrent ovarian cancer remains a high priority. Using expression profiling, we and others have recently found claudin-3 and claudin-4 genes to be highly expressed in ovarian cancer. Because these tight junction proteins have been described as the low- and high-affinity receptors, respectively, for the cytotoxic Clostridium perfringens enterotoxin (CPE), in this study we investigated the level of expression of claudin-3 and/or claudin-4 in chemotherapy-naive and chemotherapy-resistant primary human ovarian cancers as well as their sensitivity to CPE treatment in vitro. We report that 100% (17 of 17) of the primary ovarian tumors tested overexpress one or both CPE receptors by quantitative reverse transcription-PCR. All ovarian tumors showed a dose dependent cytotoxic effect to CPE in vitro. Importantly, chemotherapy resistant/recurrent ovarian tumors were found to express claudin-3 and claudin-4 genes at significantly higher levels when compared with chemotherapy-naive ovarian cancers. All primary ovarian tumors tested, regardless of their resistance to chemotherapeutic agents, died within 24 hours to the exposure to 3.3 microg/mL CPE in vitro. In addition, we have studied the in vivo efficacy of i.p. CPE therapy in SCID mouse xenografts in a highly relevant clinical model of chemotherapy-resistant freshly explanted human ovarian cancer (i.e., OVA-1). Multiple i.p. administration of sublethal doses of CPE every 3 days significantly inhibited tumor growth in 100% of mice harboring 1 week established OVA-1. Repeated i.p. doses of CPE also had a significant inhibitory effect on tumor progression with extended survival of animals harboring large ovarian tumor burdens (i.e., 4-week established OVA-1). Our findings suggest that CPE may have potential as a novel treatment for chemotherapy-resistant/recurrent ovarian cancer. PMID- 15899826 TI - Assessment of a combined, adenovirus-mediated oncolytic and immunostimulatory tumor therapy. AB - In this study, we identified murine breast cancer cell lines that support DNA replication of E1-deleted adenovirus vectors and which can be killed by an oncolytic adenovirus expressing adenovirus E1A and tumor necrosis factor (TNF) related apoptosis inducing ligand (TRAIL) in a replication-dependent manner (Ad.IR-E1A/TRAIL). We showed that systemic or intratumoral (i.t.) injection of adenovirus vectors into mice increases plasma levels of proinflammatory cytokines and chemokines, including TNF-alpha, INF-gamma, and MCP-1, which are potent inducers of dendritic cell maturation. Furthermore, we showed that in vivo expression of Flt3L from an adenovirus vector increases the number of CD11b+ and CD11c+ cells (populations that include dendritic cells) in the blood circulation. Based on these findings, we tested whether Ad.IR-E1A/TRAIL induced killing of tumor cells in combination with dendritic cell mobilization by Ad.Flt3L or, for comparison, Ad.GM-CSF would have an additive antitumor effect. As a model, we used immunocompetent C3H mice with syngeneic s.c. tumors derived from C3L5 cells. We found that vaccination of mice with C3L5 cells that underwent viral oncolysis in combination with Flt3L or granulocyte-macrophage colony-stimulating factor (GM CSF) expression induces a systemic antitumor immune response. I.t. injection of the oncolytic and Flt3L expressing vectors into established tumors delayed tumor growth but did not cause efficient tumor elimination. This study shows the effectiveness of a combined oncolytic/immunostimulatory tumor therapy approach. PMID- 15899827 TI - Canstatin acts on endothelial and tumor cells via mitochondrial damage initiated through interaction with alphavbeta3 and alphavbeta5 integrins. AB - Canstatin, the noncollagenous domain of collagen type IV alpha-chains, belongs to a series of collagen-derived angiogenic inhibitors. We have elucidated the functional receptors and intracellular signaling induced by canstatin that explain its strong antitumor efficacy in vivo. For this purpose, we generated a canstatin-human serum albumin (CanHSA) fusion protein, employing the HSA moiety as an expression tag. We show that CanHSA triggers a crucial mitochondrial apoptotic mechanism through procaspase-9 cleavage in both endothelial and tumor cells, which is mediated through cross-talk between alphavbeta3- and alphavbeta5 integrin receptors. As a point of reference, we employed the first three kringle domains of angiostatin (K1-3), fused with HSA, which, in contrast to CanHSA, act only on endothelial cells through alphavbeta3-integrin receptor-mediated activation of caspase-8 alone, without ensuing mitochondrial damage. Taken together, these results provide insights into how canstatin might exert its strong anticancer effect. PMID- 15899828 TI - CK2 inhibits apoptosis and changes its cellular localization following ionizing radiation. AB - In this study, we show that CK2 (casein kinase II, CKII) participates in apoptotic responses following ionizing radiation (IR). Using HeLa human cervical carcinoma cells, we find that transfection of small interfering RNA against the CK2 alpha and/or alpha' catalytic subunits results in enhanced apoptosis following IR damage as measured by flow cytometry techniques, compared with a control small interfering RNA. Within 2 to 6 hours of IR, CK2 alpha partially localizes to perinuclear structures, whereas a marked nuclear localization of alpha' occurs. Treatment with a pan-caspase inhibitor or transfection of ARC (apoptosis repressor with caspase recruitment domain) suppresses the apoptotic response to IR in the CK2-reduced cells, indicating involvement of caspases. Additionally, we find that CK2 alpha and/or alpha' reduction affects cell cycle progression independent of IR damage in this human cell line. However, the G2-M checkpoint following IR is not affected in CK2 alpha- and/or alpha'-reduced cells. Thus, our data suggest that CK2 participates in inhibition of apoptosis and negatively regulates caspase activity following IR damage. PMID- 15899829 TI - Inhibition of the DNA-dependent protein kinase catalytic subunit radiosensitizes malignant glioma cells by inducing autophagy. AB - DNA-dependent protein kinase (DNA-PK) plays a major role in the repair of DNA double-strand breaks induced by ionizing radiation (IR). Lack of DNA-PK causes defective DNA double-strand break repair and radiosensitization. In general, the cell death induced by IR is considered to be apoptotic. On the other hand, nonapoptotic cell death, autophagy, has recently attracted attention as a novel response of cancer cells to chemotherapy and IR. Autophagy is a protein degradation system characterized by a prominent formation of double-membrane vesicles in the cytoplasm. Little is known, however, regarding the relationship between DNA-PK and IR-induced autophagy. In the present study, we used human malignant glioma M059J and M059K cells to investigate the role of DNA-PK in IR induced apoptotic and autophagic cell death. Low-dose IR induced massive autophagic cell death in M059J cells that lack the catalytic subunit of DNA-PK (DNA-PKcs). Most M059K cells, the counterpart of M059J cells in which DNA-PKcs are expressed at normal levels, survived, and proliferated although a small portion of the cells underwent apoptosis. Low-dose IR inhibited the phosphorylation of p70(S6K), a molecule downstream of the mammalian target of rapamycin associated with autophagy in M059J cells but not in M059K cells. The treatment of M059K cells with antisense oligonucleotides against DNA-PKcs caused radiation-induced autophagy and radiosensitized the cells. Furthermore, antisense oligonucleotides against DNA-PKcs radiosensitized other malignant glioma cell lines with DNA-PK activity, U373-MG and T98G, by inducing autophagy. The specific inhibition of DNA-PKcs may be promising as a new therapy to radiosensitize malignant glioma cells by inducing autophagy. PMID- 15899830 TI - Immune selection and emergence of aggressive tumor variants as negative consequences of Fas-mediated cytotoxicity and altered IFN-gamma-regulated gene expression. AB - Antitumor responses can be induced in patients via active or adoptive immunotherapy, yet complete tumor eradication occurs infrequently. This paradox in tumor immunology led us to address two questions: (a) Does an antitumor response, which is intended to destroy the aberrant target population, also at the same time select for aggressive tumor variants (ATV) in vivo? (b) If this process does occur, what is the contribution of the perforin- or Fas-mediated effector mechanism in the immune selection of such ATV? Here, in an experimental mouse lung metastasis model, we showed that ATV generated either naturally in vivo or in vitro by anti-Fas selection resembled each other biologically and genetically as judged by enhanced tumor growth and genome-scale gene expression profiling, respectively. Furthermore, ATV that survived CTL adoptive immunotherapy displayed an even more profound loss of Fas expression and function as well as enhanced malignant proficiency in vivo. ATV, however, retained sensitivity to perforin-mediated lysis in vitro. Lastly, such ATV displayed a diminished responsiveness in their expression of IFN-gamma-regulated genes, including those mechanistically linked to Fas-mediated death (i.e., Fas and caspase-1). Overall, we showed that (a) immune selection did occur in vivo and played an important role in the emergence of ATV, (b) ATV bearing a Fas-resistant phenotype was a chief consequence of immune selection, and (c) an overall diminished responsiveness of IFN-gamma-regulated gene expression was characteristic of ATV. Thus, in this model, Fas-mediated cytotoxicity, in concert with IFN-gamma-regulated gene expression, mechanistically constituted significant determinants of immune selection of ATV in vivo. PMID- 15899831 TI - AZD2171: a highly potent, orally bioavailable, vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor for the treatment of cancer. AB - Inhibition of vascular endothelial growth factor-A (VEGF) signaling is a promising therapeutic approach that aims to stabilize the progression of solid malignancies by abrogating tumor-induced angiogenesis. This may be accomplished by inhibiting the kinase activity of VEGF receptor-2 (KDR), which has a key role in mediating VEGF-induced responses. The novel indole-ether quinazoline AZD2171 is a highly potent (IC50 < 1 nmol/L) ATP-competitive inhibitor of recombinant KDR tyrosine kinase in vitro. Concordant with this activity, in human umbilical vein endothelial cells, AZD2171 inhibited VEGF-stimulated proliferation and KDR phosphorylation with IC50 values of 0.4 and 0.5 nmol/L, respectively. In a fibroblast/endothelial cell coculture model of vessel sprouting, AZD2171 also reduced vessel area, length, and branching at subnanomolar concentrations. Once daily oral administration of AZD2171 ablated experimental (VEGF-induced) angiogenesis in vivo and inhibited endochondral ossification in bone or corpora luteal development in ovary; physiologic processes that are highly dependent upon neovascularization. The growth of established human tumor xenografts (colon, lung, prostate, breast, and ovary) in athymic mice was inhibited dose-dependently by AZD2171, with chronic administration of 1.5 mg per kg per day producing statistically significant inhibition in all models. A histologic analysis of Calu 6 lung tumors treated with AZD2171 revealed a reduction in microvessel density within 52 hours that became progressively greater with the duration of treatment. These changes are indicative of vascular regression within tumors. Collectively, the data obtained with AZD2171 are consistent with potent inhibition of VEGF signaling, angiogenesis, neovascular survival, and tumor growth. AZD2171 is being developed clinically as a once-daily oral therapy for the treatment of cancer. PMID- 15899832 TI - Selective killing of adriamycin-resistant (G2 checkpoint-deficient and MRP1 expressing) cancer cells by docetaxel. AB - Chemotherapy of cancer is limited by toxicity to normal cells. Drug resistance further limits the therapy. Here, we investigated selective killing of drug resistant cancer cells by antagonistic drug combinations, which can spare (because of drug antagonism) normal cells. We used paired cell lines that are resistant to Adriamycin due to either expression of MRP1 or lack of G2 checkpoints. The goal was to selectively kill Adriamycin-resistant cancer cells with Docetaxel (Taxotere), while protecting parental (Adriamycin-sensitive) cells, using cytostatic concentrations of Adriamycin. Taxotere kills cells in mitosis. Therefore, by arresting parental cells in G2, 20 to 40 ng/mL of Adriamycin prevented cell death caused by Taxotere. Also, Adriamycin prevented the effects of Taxotere in normal human lymphocytes. In contrast, Taxotere selectively killed MRP1-expressing leukemia cells, which did not undergo G2 arrest in the presence of Adriamycin. Also, in the presence of Adriamycin, HCT116 p21-/- cancer cells with a defective G2 checkpoint entered mitosis and were selectively killed by Taxotere. Finally, 20 ng/mL of Adriamycin protected normal FDC-P1 hematopoietic cells from Taxotere. Whereas parental cells were protected by Adriamycin, the mitogen-activated protein/extracellular signal-regulated kinase inhibitor PD90598 potentiated the cytotoxic effect of Taxotere selectively in Raf-1-transformed FDC-P1 leukemia cells. We propose a therapeutic strategy to prevent normal cells from entering mitosis while increasing apoptosis selectively in mitotic cancer cells. PMID- 15899833 TI - A monoclonal antibody that binds anionic phospholipids on tumor blood vessels enhances the antitumor effect of docetaxel on human breast tumors in mice. AB - Anionic phospholipids, principally phosphatidylserine, become exposed on the external surface of viable vascular endothelial cells in tumors, providing an excellent marker for tumor vascular targeting. We recently raised an IgG monoclonal antibody, 3G4, which binds to anionic phospholipids in a beta2 glycoprotein I-dependent manner. It inhibited tumor growth in a variety of rodent tumor models by stimulating antibody-dependent cellular cytotoxicity toward tumor vessels. In the present study, we tested the hypothesis that docetaxel, which is known to have antivascular effects on tumors, might induce exposure of anionic phospholipids on tumor vasculature and, thus, enhance the antitumor activity of 3G4. Treatment of human umbilical vascular endothelial cells with subtoxic concentrations of docetaxel (20 pmol/L) in vitro caused anionic phospholipids to be externalized without inducing apoptosis. Docetaxel treatment of mice increased the percentage of tumor vessels that expose anionic phospholipids from 35% to 60%. No induction of phosphatidylserine was observed on vessels in normal tissues even after systemic treatment with docetaxel. Treatment of mice bearing orthotopic MDA-MB-435 human breast tumors with 3G4 plus docetaxel inhibited tumor growth by 93%. Treatment of mice bearing disseminated MDA-MB-435 tumors with 3G4 plus docetaxel reduced the average number of tumor colonies in the lungs by 93% and half the animals did not develop tumors. In both tumor models, the antitumor effect of the combination was statistically superior (P < 0.01) to that of docetaxel or 3G4 alone. Combination therapy reduced the tumor vessel density and plasma volume in tumors to a greater extent than did the individual drugs. The combination therapy was no more toxic to the mice than was docetaxel alone. These results indicate that, as an adjuvant therapy, 3G4 could enhance the therapeutic efficacy of docetaxel in breast cancer patients. PMID- 15899834 TI - 1'-acetoxychavicol acetate is a novel nuclear factor kappaB inhibitor with significant activity against multiple myeloma in vitro and in vivo. AB - 1'-Acetoxychavicol acetate (ACA) is a component of a traditional Asian condiment obtained from the rhizomes of the commonly used ethno-medicinal plant Languas galanga. Here, we show for the first time that ACA dramatically inhibits the cellular growth of human myeloma cells via the inhibition of nuclear factor kappaB (NF-kappaB) activity. In myeloma cells, cultivation with ACA induced G0-G1 phase cell cycle arrest, followed by apoptosis. Treatment with ACA induced caspase 3, 9, and 8 activities, suggesting that ACA-induced apoptosis in myeloma cells mediates both mitochondrial- and Fas-dependent pathways. Furthermore, we showed that ACA significantly inhibits the serine phosphorylation and degradation of IkappaBalpha. ACA rapidly decreased the nuclear expression of NF-kappaB, but increased the accumulation of cytosol NF-kappaB in RPMI8226 cells, indicating that ACA inhibits the translocation of NF-kappaB from the cytosol to the nucleus. To evaluate the effects of ACA in vivo, RPMI8226-transplanted NOD/SCID mice were treated with ACA. Tumor weight significantly decreased in the ACA-treated mice compared with the control mice. In conclusion, ACA has an inhibitory effect on NF kappaB, and induces the apoptosis of myeloma cells in vitro and in vivo. ACA, therefore, provides a new biologically based therapy for the treatment of multiple myeloma patients as a novel NF-kappaB inhibitor. PMID- 15899835 TI - The human multidrug resistance protein MRP5 transports folates and can mediate cellular resistance against antifolates. AB - Members of the multidrug resistance protein family, notably MRP1-4/ABCC1-4, and the breast cancer resistance protein BCRP/ABCG2 have been recognized as cellular exporters for the folate antagonist methotrexate (MTX). Here we show that MRP5/ABCC5 is also an antifolate and folate exporter based on the following evidence: (a) Using membrane vesicles from HEK293 cells, we show that MRP5 transports both MTX (KM = 1.3 mmol/L and VMAX = 780 pmol per mg protein per minute) and folic acid (KM = 1.0 mmol/L and VMAX = 875 pmol per mg protein per minute). MRP5 also transports MTX-glu2 (KM = 0.7 mmol/L and VMAX = 450 pmol per mg protein per minute) but not MTX-glu3. (b) Both accumulation of total [3H]MTX and of MTX polyglutamates were significantly reduced in MRP5 overexpressing cells. (c) Cell growth inhibition studies with MRP5 transfected HEK293 cells showed that MRP5 conferred high-level resistance (>160-fold) against the antifolates MTX, GW1843, and ZD1694 (raltitrexed) in short-term (4 hours) incubations with high drug concentrations; this resistance was proportional to the MRP5 level. (d) MRP5-mediated resistance (8.5- and 2.1-fold) was also found in standard long-term incubations (72 hours) at low concentrations of ZD1694 and GW1843. These results show the potential of MRP5 to mediate transport of (anti)folates and contribute to resistance against antifolate drugs. PMID- 15899836 TI - Enhancement of folate receptor alpha expression in tumor cells through the glucocorticoid receptor: a promising means to improved tumor detection and targeting. AB - The utility of the folate receptor (FR) type alpha, in a broad range of targeted therapies and as a diagnostic serum marker in cancer, is confounded by its variable tumor expression levels. FR-alpha, its mRNA and its promoter activity were coordinately up-regulated by the glucocorticoid receptor (GR) agonist, dexamethasone. Optimal promoter activation which occurred at <50 nmol/L dexamethasone was inhibited by the GR antagonist, RU486, and was enhanced by coactivators, supporting GR mediation of the dexamethasone effect. The dexamethasone response of the FR-alpha promoter progressed even after dexamethasone was withdrawn, but this delayed effect required prior de novo protein synthesis indicating an indirect regulation. The dexamethasone effect was mediated by the G/C-rich (Sp1 binding) element in the core P4 promoter and was optimal in the proper initiator context without associated changes in the complement of major Sp family proteins. Histone deacetylase (HDAC) inhibitors potentiated dexamethasone induction of FR-alpha independent of changes in GR levels. Dexamethasone/HDAC inhibitor treatment did not cause de novo FR-alpha expression in a variety of receptor-negative cells. In a murine HeLa cell tumor xenograft model, dexamethasone treatment increased both tumor-associated and serum FR-alpha. The results support the concept of increasing FR-alpha expression selectively in the receptor-positive tumors by brief treatment with a nontoxic dose of a GR agonist, alone or in combination with a well-tolerated HDAC inhibitor, to increase the efficacy of various FR-alpha-dependent therapeutic and diagnostic applications. They also offer a new paradigm for cancer diagnosis and combination therapy that includes altering a marker or a target protein expression using general transcription modulators. PMID- 15899837 TI - Omega-3 polyunsaturated fatty acids regulate syndecan-1 expression in human breast cancer cells. AB - Human epidemiologic studies and animal model studies support a role for n-3 polyunsaturated fatty acids (n-3 PUFA) in prevention or inhibition of breast cancer. However, mechanisms for this protection remain unclear. Syndecan-1 is a heparan sulfate proteoglycan, expressed on the surface of mammary epithelial cells and known to regulate many biological processes, including cytoskeletal organization, growth factor signaling, and cell-cell adhesion. We studied effects of n-3 PUFA on syndecan-1 expression in human mammary cell lines. PUFA were delivered to cells by low-density lipoproteins (LDL) isolated from the plasma of monkeys fed diets enriched in fish oil (n-3 PUFA) or linoleic acid (n-6 PUFA). Proteoglycan synthesis was measured by incorporation of [35S]-sodium sulfate. No effect of either LDL was observed in nontumorigenic MCF-10A cells, whereas in MCF 7 breast cancer cells, treatment with n-3-enriched LDL but not n-6-enriched LDL resulted in significantly greater synthesis of a proteoglycan identified by immunoprecipitation as syndecan-1. Using real-time reverse transcription-PCR (RT PCR), it was shown that n-3-enriched LDL significantly increased the expression of syndecan-1 mRNA in a dose-dependent manner and maximal effective time at 8 hours of treatment. The effect was mimicked by an agonist for peroxisome proliferator-activated receptor gamma (PPARgamma) and eliminated by the presence of PPARgamma antagonist suggesting a role for PPARgamma in syndecan enhancement. Our studies show that n-3 LDL modifies the production of syndecan-1 in human breast cancer cells and suggest that biological processes regulated by syndecan-1 may be modified through LDL delivery of n-3 PUFA. PMID- 15899838 TI - Milk thistle and prostate cancer: differential effects of pure flavonolignans from Silybum marianum on antiproliferative end points in human prostate carcinoma cells. AB - Extracts from the seeds of milk thistle, Silybum marianum, are known commonly as silibinin and silymarin and possess anticancer actions on human prostate carcinoma in vitro and in vivo. Seven distinct flavonolignan compounds and a flavonoid have been isolated from commercial silymarin extracts. Most notably, two pairs of diastereomers, silybin A and silybin B and isosilybin A and isosilybin B, are among these compounds. In contrast, silibinin is composed only of a 1:1 mixture of silybin A and silybin B. With these isomers now isolated in quantities sufficient for biological studies, each pure compound was assessed for antiproliferative activities against LNCaP, DU145, and PC3 human prostate carcinoma cell lines. Isosilybin B was the most consistently potent suppressor of cell growth relative to either the other pure constituents or the commercial extracts. Isosilybin A and isosilybin B were also the most effective suppressors of prostate-specific antigen secretion by androgen-dependent LNCaP cells. Silymarin and silibinin were shown for the first time to suppress the activity of the DNA topoisomerase IIalpha gene promoter in DU145 cells and, among the pure compounds, isosilybin B was again the most effective. These findings are significant in that isosilybin B composes no more than 5% of silymarin and is absent from silibinin. Whereas several other more abundant flavonolignans do ultimately influence the same end points at higher exposure concentrations, these findings are suggestive that extracts enriched for isosilybin B, or isosilybin B alone, might possess improved potency in prostate cancer prevention and treatment. PMID- 15899839 TI - Phenotypic profiling of engineered mouse melanomas with manipulated histamine production identifies histamine H2 receptor and rho-C as histamine-regulated melanoma progression markers. AB - In the present study, the impact of acquired neoplastic L-histidine decarboxylase (HDC) expression, and its direct consequence, the release of histamine in the tumor environment, was assessed on melanoma tumor progression. B16-F10 mouse melanoma cells were manipulated via stable transfection, and nine novel transgenic variants were generated in triplicates, constitutively expressing the full-length sense mouse HDC mRNA, a mock control, and an antisense HDC RNA segment, respectively. Establishing both primary skin tumors and lung metastases in C57BL/6 mice, the nine variants with different histamine-releasing capacities were subjected to a comprehensive comparative progression profiling in vivo. Our analyses showed trends of markedly accelerated tumor growth (P < 0.001), and moderately increased metastatic colony-forming potential (P = 0.010) along with rising levels of local histamine production. Using RNase protection assay for screening of the melanoma progression profile, and Western blotting for subsequent result validation, we looked for molecular progression markers affected by melanoma histamine secretion. Investigation of 21 functionally clustered markers associated with tumor proliferation, angiogenesis, invasivity, metastasis formation, local or systemic immunomodulation, and histamine signaling revealed positive correlations between histamine production, tumor histamine H2 receptor and rho-C expression (P < 0.001, P = 0.002, respectively). These observations confirm the involvement of histamine in the molecular machinery of melanoma progression. PMID- 15899840 TI - TReP-132 controls cell proliferation by regulating the expression of the cyclin dependent kinase inhibitors p21WAF1/Cip1 and p27Kip1. AB - The transcriptional regulating protein of 132 kDa (TReP-132) has been identified in steroidogenic tissues, where it acts as a coactivator of steroidogenic factor 1 (SF-1). We show here that TReP-132 plays a role in the control of cell proliferation. In human HeLa cells, TReP-132 knockdown by using small interfering RNA resulted in increased G(1)-->S cell cycle progression. The growth-inhibitory effects of TReP-132 was further shown to be mediated by induction of G(1) cyclin dependent kinase inhibitors p21(WAF1) (p21) and p27(KIP1) (p27) expression levels. As a consequence, G(1) cyclin/cyclin-dependent kinase activities and pRB phosphorylation were markedly reduced, and cell cycle progression was blocked in the G(1) phase. The stimulatory effect of TReP-132 on p21 and p27 gene transcription involved interaction of TReP-132 with the transcription factor Sp1 at proximal Sp1-binding sites in their promoters. Moreover, in different breast tumor cell lines, endogenous TReP-132 expression was positively related with a lower proliferation rate. In addition, TReP-132 knockdown resulted in enhanced cell proliferation and lowered p21 and p27 mRNA levels in the steroid-responsive and nonresponsive T-47D and MDA-MB-231 cell lines, respectively. Finally, a statistic profiling of human breast tumor samples highlighted that expression of TReP-132 is correlated with p21 and p27 levels and is associated with lower tumor incidence and aggressiveness. Together, these results identify TReP-132 as a basal cell cycle regulatory protein acting, at least in part, by interacting with Sp1 to activate the p21 and p27 gene promoters. PMID- 15899841 TI - CSN5/Jab1 is involved in ligand-dependent degradation of estrogen receptor {alpha} by the proteasome. AB - Here, we show that estrogen receptor alpha (ERalpha) coimmunoprecipitates with CSN5/Jab1, a subunit of the COP9 signalosome (CSN), and that overexpression of CSN5/Jab1 causes an increase in ligand-induced ERalpha degradation. Inhibition of either the kinase activity associated with the CSN complex by curcumin or of nuclear export by leptomycin B (LMB) impaired estradiol-induced ERalpha degradation by the proteasome. Degradation of ERalpha induced by the pure antagonist ICI 182,780 (ICI) was blocked by curcumin but not by LMB, indicating that in the presence of ICI, ERalpha is degraded by a nuclear fraction of the proteasome. In addition, we observed that curcumin inhibited estradiol-induced phosphorylation of ERalpha. The use of three inhibitors of ERalpha degradation that target different steps of the estrogen response pathway (inhibition of the CSN-associated kinase, nuclear export, and proteasome) suggests that a phosphorylation event inhibited by curcumin is necessary for ERalpha binding to its cognate DNA target. Our results demonstrate that transcription per se is not required for ERalpha degradation and that assembly of the transcription initiation complex is sufficient to target ERalpha for degradation by the proteasome. PMID- 15899842 TI - Trm11p and Trm112p are both required for the formation of 2-methylguanosine at position 10 in yeast tRNA. AB - N(2)-Monomethylguanosine-10 (m(2)G10) and N(2),N(2)-dimethylguanosine-26 (m(2)(2)G26) are the only two guanosine modifications that have been detected in tRNA from nearly all archaea and eukaryotes but not in bacteria. In Saccharomyces cerevisiae, formation of m(2)(2)G26 is catalyzed by Trm1p, and we report here the identification of the enzymatic activity that catalyzes the formation of m(2)G10 in yeast tRNA. It is composed of at least two subunits that are associated in vivo: Trm11p (Yol124c), which is the catalytic subunit, and Trm112p (Ynr046w), a putative zinc-binding protein. While deletion of TRM11 has no detectable phenotype under laboratory conditions, deletion of TRM112 leads to a severe growth defect, suggesting that it has additional functions in the cell. Indeed, Trm112p is associated with at least four proteins: two tRNA methyltransferases (Trm9p and Trm11p), one putative protein methyltransferase (Mtc6p/Ydr140w), and one protein with a Rossmann fold dehydrogenase domain (Lys9p/Ynr050c). In addition, TRM11 interacts genetically with TRM1, thus suggesting that the absence of m(2)G10 and m(2)(2)G26 affects tRNA metabolism or functioning. PMID- 15899843 TI - Mouse axin and axin2/conductin proteins are functionally equivalent in vivo. AB - Axin is a central component of the canonical Wnt signal transduction machinery, serving as a scaffold for the beta-catenin destruction complex. The related protein Axin2/Conductin, although less extensively studied, is thought to perform similar functions. Loss of Axin causes early embryonic lethality, while Axin2 null mice are viable but have craniofacial defects. Mutations in either gene contribute to cancer in humans. The lack of redundancy between Axin and Axin2 could be due to their different modes of expression: while Axin is expressed ubiquitously, Axin2 is expressed in tissue- and developmental-stage-specific patterns, and its transcription is induced by canonical Wnt signaling. Alternatively, the two proteins might have partially different functions, a hypothesis supported by the observation that they differ in their subcellular localizations in colon epithelial cells. To test the functional equivalence of Axin and Axin2 in vivo, we generated knockin mice in which the Axin gene was replaced with Myc-tagged Axin or Axin2 cDNA. Mice homozygous for the resulting alleles, Axin(Ax) or Axin(Ax2), express no endogenous Axin but express either Myc Axin or Myc-Axin2 under the control of the Axin locus. Both Axin(Ax/Ax) and Axin(Ax2/Ax2) homozygotes are apparently normal and fertile, demonstrating that the Axin and Axin2 proteins are functionally equivalent. PMID- 15899844 TI - Fission yeast rad51 and dmc1, two efficient DNA recombinases forming helical nucleoprotein filaments. AB - Homologous recombination is important for the repair of double-strand breaks during meiosis. Eukaryotic cells require two homologs of Escherichia coli RecA protein, Rad51 and Dmc1, for meiotic recombination. To date, it is not clear, at the biochemical level, why two homologs of RecA are necessary during meiosis. To gain insight into this, we purified Schizosaccharomyces pombe Rad51 and Dmc1 to homogeneity. Purified Rad51 and Dmc1 form homo-oligomers, bind single-stranded DNA preferentially, and exhibit DNA-stimulated ATPase activity. Both Rad51 and Dmc1 promote the renaturation of complementary single-stranded DNA. Importantly, Rad51 and Dmc1 proteins catalyze ATP-dependent strand exchange reactions with homologous duplex DNA. Electron microscopy reveals that both S. pombe Rad51 and Dmc1 form nucleoprotein filaments. Rad51 formed helical nucleoprotein filaments on single-stranded DNA, whereas Dmc1 was found in two forms, as helical filaments and also as stacked rings. These results demonstrate that Rad51 and Dmc1 are both efficient recombinases in lower eukaryotes and reveal closer functional and structural similarities between the meiotic recombinase Dmc1 and Rad51. The DNA strand exchange activity of both Rad51 and Dmc1 is most likely critical for proper meiotic DNA double-strand break repair in lower eukaryotes. PMID- 15899845 TI - The thymine DNA glycosylase MBD4 represses transcription and is associated with methylated p16(INK4a) and hMLH1 genes. AB - Epigenetic silencing through methyl-CpG (mCpG) is implicated in many biological patterns such as genome imprinting, X chromosome inactivation, and cancer development. In this process, the mCpG binding domain (MBD) proteins play an essential role in transmitting epigenetic information to downstream regulatory proteins. Among the five MBD proteins identified so far, MBD4 has been the only exception; it has long been thought to be a DNA repair protein. Herein we demonstrate that MBD4 has the ability to repress transcription through mCpG. Transcriptional repression by the MBD4 is histone deacetylase (HDAC) dependent, and MBD4 directly binds to Sin3A and HDAC1 at three central regions that overlap transcriptional repression domains. Furthermore, a chromatin immunoprecipitation assay clearly shows that MBD4 binds to hypermethylated promoters of the p16(INK4a) and hMLH1 genes. These results suggest that MBD4 is one of the essential components involved in epigenetic silencing in cancer and its repair activity is necessary for the maintenance of hypermethylated promoters. PMID- 15899846 TI - Retroviral splicing suppressor sequesters a 3' splice site in a 50S aberrant splicing complex. AB - Retroviral replication requires both spliced and unspliced mRNAs. Splicing suppression of avian retroviral RNA depends in part upon a cis-acting element within the gag gene called the negative regulator of splicing (NRS). The NRS, linked to a downstream intron and exon (NRS-Ad3'), was not capable of splicing in vitro. However, a double-point mutation in the NRS pseudo-5' splice site sequence converted it into a functional 5' splice site. The wild-type (WT) NRS-Ad3' transcript assembled an approximately 50S spliceosome-like complex in vitro; its sedimentation rate was similar to that of a functional spliceosome formed on the mutant NRS-Ad3' RNA. The five major spliceosomal snRNPs were observed in both complexes by affinity selection. In addition, U11 snRNP was present only in the WT NRS-Ad3' complex. Addition of heparin to these complexes destabilized the WT NRS-Ad3' complex; it was incapable of forming a B complex on a native gel. Furthermore, the U5 snRNP protein, hPrp8, did not cross-link to the NRS pseudo-5' splice site, suggesting that the tri-snRNP complex was not properly associated with it. We propose that this aberrant, stalled spliceosome, containing U1, U2, and U11 snRNPs and a loosely associated tri-snRNP, sequesters the 3' splice site and prevents its interaction with the authentic 5' splice site upstream of the NRS. PMID- 15899847 TI - Recombination at long mutant telomeres produces tiny single- and double-stranded telomeric circles. AB - Recombinational telomere elongation (RTE) known as alternate lengthening of telomeres is the mechanism of telomere maintenance in up to 5 to 10% of human cancers. The telomeres of yeast mutants lacking telomerase can also be maintained by recombination. Previously, we proposed the roll-and-spread model to explain this elongation in the yeast Kluveromyces lactis. This model suggests that a very small ( approximately 100-bp) circular molecule of telomeric DNA is copied by a rolling circle event to generate a single long telomere. The sequence of this primary elongated telomere is then spread by recombination to all remaining telomeres. Here we show by two-dimensional gel analysis and electron microscopy that small circles of single- and double-stranded telomeric DNA are commonly made by recombination in a K. lactis mutant with long telomeres. These circles were found to be especially abundant between 100 and 400 bp (or nucleotides). Interestingly, the single-stranded circles consist of only the G-rich telomeric strand sequence. To our knowledge this is the first report of single-stranded telomeric circles as a product of telomere dysfunction. We propose that the small telomeric circles form through the resolution of an intratelomeric strand invasion which resembles a t-loop. Our data reported here demonstrate that K. lactis can, in at least some circumstances, make telomeric circles of the very small sizes predicted by the roll-and-spread model. The very small circles seen here are both predicted products of telomere rapid deletion, a process observed in both human and yeast cells, and predicted templates for roll-and-spread RTE. PMID- 15899848 TI - Determinants of HMGB proteins required to promote RAG1/2-recombination signal sequence complex assembly and catalysis during V(D)J recombination. AB - Efficient assembly of RAG1/2-recombination signal sequence (RSS) DNA complexes that are competent for V(D)J cleavage requires the presence of the nonspecific DNA binding and bending protein HMGB1 or HMGB2. We find that either of the two minimal DNA binding domains of HMGB1 is effective in assembling RAG1/2-RSS complexes on naked DNA and stimulating V(D)J cleavage but that both domains are required for efficient activity when the RSS is incorporated into a nucleosome. The single-domain HMGB protein from Saccharomyces cerevisiae, Nhp6A, efficiently assembles RAG1/2 complexes on naked DNA; however, these complexes are minimally competent for V(D)J cleavage. Nhp6A forms much more stable DNA complexes than HMGB1, and a variety of mutations that destabilize Nhp6A binding to bent microcircular DNA promote increased V(D)J cleavage. One of the two DNA bending wedges on Nhp6A and the analogous phenylalanine wedge at the DNA exit site of HMGB1 domain A were found to be essential for promoting RAG1/2-RSS complex formation. Because the phenylalanine wedge is required for specific recognition of DNA kinks, we propose that HMGB proteins facilitate RAG1/2-RSS interactions by recognizing a distorted DNA structure induced by RAG1/2 binding. The resulting complex must be sufficiently dynamic to enable the series of RAG1/2-mediated chemical reactions on the DNA. PMID- 15899849 TI - A diacylglycerol-protein kinase C-RasGRP1 pathway directs Ras activation upon antigen receptor stimulation of T cells. AB - Ras GTPases are on/off switches regulating numerous cellular responses by signaling to various effector molecules. In T lymphocytes, Ras can be activated by two Ras exchange factors, SOS and RasGRP1, which are recruited through the adapters Grb2 and LAT and via the second-messenger diacylglycerol (DAG), respectively. Mitogen-activated protein (MAP) kinase phosphorylation patterns induced by active Ras can vary and contribute to distinct cellular responses. The different consequences of Ras activation by either guanine exchange factor are unknown. DAG also recruits and activates the kinase protein kinase Ctheta (PKCtheta) turning on the Erk MAP kinase pathway, but the biochemical mechanism responsible is unclear. We generated T-cell clones deficient in phorbol myristate acetate (a surrogate for DAG)-induced Ras activation. Analysis of a RasGRP1 deficient Jurkat T-cell clone and RasGRP1 RNA interference in wild-type cells revealed that RasGRP1 is required for optimal, antigen receptor-triggered Ras-Erk activation. RasGRP1 relies on its DAG-binding domain to selectively activate Erk kinases. Activation of Erk correlates with the phosphorylation of threonine residue 184 in RasGRP1. This phosphorylation event requires the activities of novel PKC kinases. Conversely, active PKCtheta depends on RasGRP1 sufficiency to effectively trigger downstream events. Last, DAG-PKC-RasGRP1-driven Ras-Erk activation in T cells is a unique signaling event, not simply compensated for by SOS activity. PMID- 15899850 TI - Biological and biochemical functions of RNA in the tetrahymena telomerase holoenzyme. AB - Telomerase extends chromosome ends by the synthesis of tandem simple-sequence repeats. Studies of minimal recombinant telomerase ribonucleoprotein (RNP) reconstituted in vitro have revealed sequences within the telomerase RNA subunit (TER) that are required to establish its internal template and other unique features of enzyme activity. Here we test the significance of these motifs following TER assembly into telomerase holoenzyme in vivo. We established a method for stable expression of epitope-tagged TER and TER variants in place of wild-type Tetrahymena TER. We found that sequence substitutions in nontemplate regions of TER altered telomere length maintenance in vivo, with an increase or decrease in the set point for telomere length homeostasis. We also characterized the in vitro activity of the telomerase holoenzymes reconstituted with TER variants, following RNA-based RNP affinity purification from cell extracts. We found that nontemplate sequence substitutions imposed specific defects in the fidelity and processivity of template use. These findings demonstrate nontemplate functions of TER that are critical for the telomerase holoenzyme catalytic cycle and for proper telomere length maintenance in vivo. PMID- 15899851 TI - Single and combined deletions of the NTAL/LAB and LAT adaptors minimally affect B cell development and function. AB - NTAL (non-T-cell activation linker, also called LAB) and LAT (linker for activation of T cells) are evolutionarily related transmembrane adaptor proteins that are phosphorylated upon immunoreceptor engagement. Using quantitative reverse transcription-PCR, both NTAL and LAT were found to be expressed in B cells. However, LAT expression was limited to early B cells, whereas NTAL expression typified mature B cells. To delineate their roles in B-cell development and function, Ntal-deficient mice were generated and crossed with Lat deficient mice. B cells developed in Lat(-/-) Ntal(-/-) double-deficient mice and in mice lacking either of the two adaptors with the same efficiency as in wild type mice. Upon B-cell antigen receptor cross-linking, Ntal(-/-) B cells exhibited slightly increased Ca(2+) mobilization and proliferation. In addition, Ntal-deficient mice had increased levels of natural antibodies and slightly increased humoral response to a T-dependent antigen. Normal titers of serum specific immunoglobulins were produced in response to a T-cell-independent antigen. Although NTAL is also expressed in plasma cells, its absence did not affect the hypergammaglobulinemia E and G1 that developed in mice with a mutation in tyrosine 136 of LAT. Therefore, NTAL does not play a role in B cells symmetric to the role played by LAT in T cells. PMID- 15899852 TI - Raf-1 serine 338 phosphorylation plays a key role in adhesion-dependent activation of extracellular signal-regulated kinase by epidermal growth factor. AB - Activation of the extracellular signal-regulated kinase (ERK) 1/2 cascade by polypeptide growth factors is tightly coupled to adhesion to extracellular matrix in nontransformed cells. Raf-1, the initial kinase in this cascade, is intricately regulated by phosphorylation, localization, and molecular interactions. We investigated the complex interactions between Raf-1, protein kinase A (PKA), and p21-activated kinase (PAK) to determine their roles in the adhesion dependence of signaling from epidermal growth factor (EGF) to ERK. We conclude that Raf-1 phosphorylation on serine 338 (S338) is a critical step that is inhibited in suspended cells. Restoration of phosphorylation at S338, either by expression of highly active PAK or by expression of an S338 phospho-mimetic Raf-1 mutation, led to a partial rescue of ERK activation in suspended cells. Raf 1 inhibition in suspension was not due to excessive negative regulation on inhibitory sites S43 and S259, as these serines were largely dephosphorylated in suspended cells. Finally, strong phosphorylation of Raf-1 S338 provided resistance to PKA-mediated inhibition of ERK activation. Phosphorylation at Raf-1 S43 and S259 by PKA only weakly inhibited EGF activation of Raf-1 and ERK when cells maintained high Raf-1 S338 phosphorylation. PMID- 15899853 TI - Yeast Rmi1/Nce4 controls genome stability as a subunit of the Sgs1-Top3 complex. AB - Genome stability requires a set of RecQ-Top3 DNA helicase-topoisomerase complexes whose sole budding yeast homolog is encoded by SGS1-TOP3. RMI1/NCE4 was identified as a potential intermediate in the SGS1-TOP3 pathway, based on the observation that strains lacking any one of these genes require MUS81 and MMS4 for viability. This idea was tested by confirming that sgs1 and rmi1 mutants display the same spectrum of synthetic lethal interactions, including the requirements for SLX1, SLX4, SLX5, and SLX8, and by demonstrating that rmi1 mus81 synthetic lethality is dependent on homologous recombination. On their own, mutations in RMI1 result in phenotypes that mimic those of sgs1 or top3 strains including slow growth, hyperrecombination, DNA damage sensitivity, and reduced sporulation. And like top3 strains, most rmi1 phenotypes are suppressed by mutations in SGS1. We show that Rmi1 forms a heteromeric complex with Sgs1-Top3 in yeast and that these proteins interact directly in a recombinant system. The Rmi1-Top3 complex is stable in the absence of the Sgs1 helicase, but the loss of either Rmi1 or Top3 in yeast compromises its partner's interaction with Sgs1. Biochemical studies demonstrate that recombinant Rmi1 is a structure-specific DNA binding protein with a preference for cruciform structures. We propose that the DNA binding specificity of Rmi1 plays a role in targeting Sgs1-Top3 to appropriate substrates. PMID- 15899854 TI - Gadd45a expression induces Bim dissociation from the cytoskeleton and translocation to mitochondria. AB - Gadd45a, a p53- and BRCA1-regulated stress protein, has been implicated in the maintenance of genomic fidelity, probably through its roles in the control of cell cycle checkpoint and apoptosis. However, the mechanism(s) by which Gadd45a is involved in the induction of apoptosis remains unclear. We show here that inducible expression of Gadd45a protein causes dissociation of Bim, a Bcl2 family member, from microtubule-associated components and translocation to mitochondria. The Bim accumulation in mitochondria enhances interaction of Bim with Bcl-2, relieves Bax from Bcl-2-bound complexes, and subsequently results in release of cytochrome c into the cytoplasm. Suppression of endogenous Bim greatly inhibits Gadd45a induction of apoptosis. Interestingly, Gadd45a interacts with elongation factor 1alpha (EF-1alpha), a microtubule-severing protein that plays an important role in maintaining cytoskeletal stability, and inhibits EF-1alpha-mediated microtubule bundling, indicating that the interaction of Gadd45a with EF-1alpha disrupts cytoskeletal stability. A mutant form of Gadd45a harboring a deletion of EF-1alpha-binding domain fails to inhibit microtubule stability and to induce Bim translocation to mitochondria. Furthermore, coexpression of EF-1alpha antagonizes Gadd45a's property of suppressing cell growth and inducing apoptosis. These findings identify a novel link that connects stress protein Gadd45a to the apoptotic machinery and address the importance of cytoskeletal stability in apoptotic response to DNA damage. PMID- 15899855 TI - Keap1 regulates the oxidation-sensitive shuttling of Nrf2 into and out of the nucleus via a Crm1-dependent nuclear export mechanism. AB - Keap1 is a negative regulator of Nrf2, a transcription factor essential for antioxidant response element (ARE)-mediated gene expression. We find that Keap1 sequesters Nrf2 in the cytoplasm, not by docking it to the actin cytoskeleton but instead through an active Crm1/exportin-dependent nuclear export mechanism. Deletion and mutagenesis studies identified a nuclear export signal (NES) in the intervening region of Keap1 comprised of hydrophobic leucine and isoleucine residues in agreement with a traditional NES consensus sequence. Mutation of the hydrophobic amino acids resulted in nuclear accumulation of both Keap1 and Nrf2, as did treatment with the drug leptomycin B, which inactivates Crm1/exportin. ARE genes were partially activated under these conditions, suggesting that additional oxidation-sensitive elements are required for full activation of the antioxidant response. Based on these data, we propose a new model for regulation of Nrf2 by Keap1. Under normal conditions, Keap1 and Nrf2 are complexed in the cytoplasm where they are targeted for degradation. Oxidative stress inactivates Keap1's NES, allowing entry of both Keap1 and Nrf2 into the nucleus and transcriptional transactivation of ARE genes. PMID- 15899856 TI - A nonhistone protein-protein interaction required for assembly of the SIR complex and silent chromatin. AB - Budding yeast silent chromatin, or heterochromatin, is composed of histones and the Sir2, Sir3, and Sir4 proteins. Their assembly into silent chromatin is believed to require the deacetylation of histones by the NAD-dependent deacetylase Sir2 and the subsequent interaction of Sir3 and Sir4 with these hypoacetylated regions of chromatin. Here we explore the role of interactions among the Sir proteins in the assembly of the SIR complex and the formation of silent chromatin. We show that significant fractions of Sir2, Sir3, and Sir4 are associated together in a stable complex. When the assembly of Sir3 into this complex is disrupted by a specific mutation on the surface of the C-terminal coiled-coil domain of Sir4, Sir3 is no longer recruited to chromatin and silencing is disrupted. Because in sir4 mutant cells the association of Sir3 with chromatin is greatly reduced despite the partial Sir2-dependent deacetylation of histones near silencers, we conclude that histone deacetylation is not sufficient for the full recruitment of silencing proteins to chromatin and that Sir-Sir interactions are essential for the assembly of heterochromatin. PMID- 15899857 TI - Endoplasmic reticulum stress induction of the Grp78/BiP promoter: activating mechanisms mediated by YY1 and its interactive chromatin modifiers. AB - The unfolded protein response is an evolutionarily conserved mechanism whereby cells respond to stress conditions that target the endoplasmic reticulum (ER). The transcriptional activation of the promoter of GRP78/BiP, a prosurvival ER chaperone, has been used extensively as an indicator of the onset of the UPR. YY1, a constitutively expressed multifunctional transcription factor, activates the Grp78 promoter only under ER stress conditions. Previously, in vivo footprinting analysis revealed that the YY1 binding site of the ER stress response element of the Grp78 promoter exhibits ER stress-induced changes in occupancy. Toward understanding the underlying mechanisms of these unique phenomena, we performed chromatin immunoprecipitation analyses, revealing that YY1 only occupies the Grp78 promoter upon ER stress and is mediated in part by the nuclear form of ATF6. We show that YY1 is an essential coactivator of ATF6 and uncover their specific interactive domains. Using small interfering RNA against YY1 and insertional mutation of the gene encoding ATF6alpha, we provide direct evidence that YY1 and ATF6 are required for optimal stress induction of Grp78. We also discovered enhancement of the ER-stressed induction of the Grp78 promoter through the interaction of YY1 with the arginine methyltransferase PRMT1 and evidence of its action through methylation of the arginine 3 residue on histone H4. Furthermore, we detected ER stress-induced binding of the histone acetyltransferase p300 to the Grp78 promoter and histone H4 acetylation. A model for the ER stress-mediated transcription factor binding and chromatin modifications at the Grp78 promoter leading to its activation is proposed. PMID- 15899858 TI - The dual-specificity phosphatase CDC14B bundles and stabilizes microtubules. AB - The Cdc14 dual-specificity phosphatases regulate key events in the eukaryotic cell cycle. However, little is known about the function of mammalian CDC14B family members. Here, we demonstrate that subcellular localization of CDC14B protein is cell cycle regulated. CDC14B can bind, bundle, and stabilize microtubules in vitro independently of its catalytic activity. Basic amino acid residues within the nucleolar targeting domain are important for both retaining CDC14B in the nucleolus and preventing microtubule bundling. Overexpression of CDC14B resulted in the formation of cytoplasmic CDC14B and microtubule bundles in interphase cells. These microtubule bundles were resistant to microtubule depolymerization reagents and enriched in acetylated alpha-tubulin. Expression of cytoplasmic forms of CDC14B impaired microtubule nucleation from the microtubule organization center. CDC14B is thus a novel microtubule-bundling and -stabilizing protein, whose regulated subcellular localization may help modulate spindle and microtubule dynamics in mitosis. PMID- 15899859 TI - In vivo HP1 targeting causes large-scale chromatin condensation and enhanced histone lysine methylation. AB - Changes in chromatin structure are a key aspect in the epigenetic regulation of gene expression. We have used a lac operator array system to visualize by light microscopy the effect of heterochromatin protein 1 (HP1) alpha (HP1alpha) and HP1beta on large-scale chromatin structure in living mammalian cells. The structure of HP1, containing a chromodomain, a chromoshadow domain, and a hinge domain, allows it to bind to a variety of proteins. In vivo targeting of an enhanced green fluorescent protein-tagged HP1-lac repressor fusion to a lac operator-containing, gene-amplified chromosome region causes local condensation of the higher-order chromatin structure, recruitment of the histone methyltransferase SETDB1, and enhanced trimethylation of histone H3 lysine 9. Polycomb group proteins of both the HPC/HPH and the EED/EZH2 complexes, which are involved in the heritable repression of gene activity, are not recruited to the amplified chromosome region by HP1alpha and HP1beta in vivo targeting. HP1alpha targeting causes the recruitment of endogenous HP1beta to the chromatin region and vice versa, indicating a direct interaction between the two HP1 homologous proteins. Our findings indicate that HP1alpha and HP1beta targeting is sufficient to induce heterochromatin formation. PMID- 15899860 TI - Decreased growth of Vhl-/- fibrosarcomas is associated with elevated levels of cyclin kinase inhibitors p21 and p27. AB - Inactivating mutations within the von Hippel-Lindau (VHL) tumor suppressor gene predispose patients to develop a variety of highly vascularized tumors. pVHL targets alpha subunits of the heterodimeric transcription factor hypoxia inducible factor (HIF), a critical regulator of energy metabolism, angiogenesis, hematopoiesis, and oxygen (O(2)) delivery, for ubiquitin-mediated degradation in an O(2)-dependent manner. To investigate the role of Vhl in cellular proliferation and tumorigenesis, we utilized mouse embryonic fibroblasts (MEFs), a common tool for analyzing cell cycle regulation, and generated Vhl(-)(/)(-) MEF derived fibrosarcomas. Surprisingly, growth of both Vhl(-)(/)(-) MEFs and fibrosarcomas was impaired, although tumor vascularity was increased. Decreased proliferation of Vhl(-)(/)(-) MEFs was correlated with an overexpression of cyclin kinase inhibitors (CKIs) p21 and p27. The transcription of p21 and p27 is inhibited by c-Myc; therefore, the induction of CKIs was attributed to the ability of HIF to antagonize c-Myc activity. Indeed, p21 mRNA levels were elevated under normoxia in Vhl(-)(/)(-) MEFs, while c-Myc transcriptional activity was markedly reduced. Gene silencing of HIF-1alpha by small interfering RNA reduced p21 and p27 protein and mRNA levels in Vhl(-)(/)(-) MEFs. The induction of p21 and p27, mediated by constitutive activation of the HIF pathway, provides a mechanism for the decreased proliferation rates of Vhl(-)(/)(-) MEFs and fibrosarcomas. These results demonstrate that a loss of pVHL can induce growth arrest in certain cells types, which suggests that additional genetic mutations are necessary for VHL-associated tumorigenesis. PMID- 15899861 TI - Mitochondrial carrier homolog 2 is a target of tBID in cells signaled to die by tumor necrosis factor alpha. AB - BID, a proapoptotic BCL-2 family member, plays an essential role in the tumor necrosis factor alpha (TNF-alpha)/Fas death receptor pathway in vivo. Activation of the TNF-R1 receptor results in the cleavage of BID into truncated BID (tBID), which translocates to the mitochondria and induces the activation of BAX or BAK. In TNF-alpha-activated FL5.12 cells, tBID becomes part of a 45-kDa cross-linkable mitochondrial complex. Here we describe the biochemical purification of this complex and the identification of mitochondrial carrier homolog 2 (Mtch2) as part of this complex. Mtch2 is a conserved protein that is similar to members of the mitochondrial carrier protein family. Our studies with mouse liver mitochondria indicate that Mtch2 is an integral membrane protein exposed on the surface of mitochondria. Using blue-native gel electrophoresis we revealed that in viable FL5.12 cells Mtch2 resides in a protein complex of ca. 185 kDa and that the addition of TNF-alpha to these cells leads to the recruitment of tBID and BAX to this complex. Importantly, this recruitment was partially inhibited in FL5.12 cells stably expressing BCL-X(L). These results implicate Mtch2 as a mitochondrial target of tBID and raise the possibility that the Mtch2-resident complex participates in the mitochondrial apoptotic program. PMID- 15899862 TI - Bim regulation of lumen formation in cultured mammary epithelial acini is targeted by oncogenes. AB - Epithelial cells organize into cyst-like structures that contain a spherical monolayer of cells that enclose a central lumen. Using a three-dimensional basement membrane culture model in which mammary epithelial cells form hollow, acinus-like structures, we previously demonstrated that lumen formation is achieved, in part, through apoptosis of centrally localized cells. We demonstrate that the proapoptotic protein Bim may selectively trigger apoptosis of the centrally localized acinar cells, leading to temporally controlled lumen formation. Bim is not detectable during early stages of three-dimensional mammary acinar morphogenesis and is then highly upregulated in all cells of acini, coincident with detection of apoptosis in the centrally localized acinar cells. Inhibition of Bim expression by RNA interference transiently blocks luminal apoptosis and delays lumen formation. Oncogenes that induce acinar luminal filling, such as ErbB2 and v-Src, suppress expression of Bim through a pathway dependent on Erk-mitogen-activated protein kinase; however, HPV 16 E7, an oncogene that stimulates cell proliferation but not luminal filling, is unable to reduce Bim expression. Thus, Bim is a critical regulator of luminal apoptosis during mammary acinar morphogenesis in vitro and may be an important target of oncogenes that disrupt glandular epithelial architecture. PMID- 15899863 TI - Tiam1-IRSp53 complex formation directs specificity of rac-mediated actin cytoskeleton regulation. AB - The exchange factor Tiam1 regulates multiple cellular functions by activating the Rac GTPase. Active Rac has various effects in cells, including alteration of actin cytoskeleton and gene expression, via binding to and modulating the activity of diverse effector proteins. How individual Rac effectors are selected for activation and regulated in response to upstream signals is not well understood. We find that Tiam1 contributes to both of these processes by binding to IRSp53, an adaptor protein that is an effector for both Rac and Cdc42. Tiam1 directs IRSp53 to Rac signaling by enhancing IRSp53 binding to both active Rac and the WAVE2 scaffold. Moreover, Tiam1 promotes IRSp53 localization to Rac induced lamellipodia rather than Cdc42-induced filopodia. Finally, IRSp53 depletion from cells prevents Tiam1-dependent lamellipodia induced by Tiam1 overexpression or platelet-derived growth factor stimulation. These findings indicate that Tiam1 not only activates Rac but also contributes to Rac signaling specificity through binding to IRSp53. PMID- 15899864 TI - Mice deficient in oocyte-specific oligoadenylate synthetase-like protein OAS1D display reduced fertility. AB - The double-stranded RNA (dsRNA)-induced interferon response is a defense mechanism against viral infection. Upon interferon activation by dsRNA, 2',5' oligoadenylate synthetase 1 (OAS1A) is induced; it binds dsRNA and converts ATP into 2',5'-linked oligomers of adenosine (called 2-5A), which activate RNase L that in turn degrades viral and cellular RNAs. In a screen to identify oocyte specific genes, we identified a novel murine cDNA encoding an ovary-specific 2',5'-oligoadenylate synthetase-like protein, OAS1D, which displays 59% identity with OAS1A. OAS1D is predominantly cytoplasmic and is exclusively expressed in growing oocytes and early embryos. Like OAS1A, OAS1D binds the dsRNA mimetic poly(I-C), but unlike OAS1A, it lacks 2'-5' adenosine linking activity. OAS1D interacts with OAS1A and inhibits the enzymatic activity of OAS1A. Mutant mice lacking OAS1D (Oas1d(-/-)) display reduced fertility due to defects in ovarian follicle development, decreased efficiency of ovulation, and eggs that are fertilized arrest at the one-cell stage. These effects are exacerbated after activation of the interferon/OAS1A/RNase L pathway by poly(I-C). We propose that OAS1D suppresses the interferon/OAS/RNase L-mediated cellular destruction by interacting with OAS1A during oogenesis and early embryonic development. PMID- 15899865 TI - YB-1 is important for late-stage embryonic development, optimal cellular stress responses, and the prevention of premature senescence. AB - Proteins containing "cold shock" domains belong to the most evolutionarily conserved family of nucleic acid-binding proteins known among bacteria, plants, and animals. One of these proteins, YB-1, is widely expressed throughout development and has been implicated as a cell survival factor that regulates the transcription and/or translation of many cellular growth and death-related genes. For these reasons, YB-1 deficiency has been predicted to be incompatible with cell survival. However, the majority of YB-1(-/-) embryos develop normally up to embryonic day 13.5 (E13.5). After E13.5, YB-1(-/-) embryos exhibit severe growth retardation and progressive mortality, revealing a nonredundant role of YB-1 in late embryonic development. Fibroblasts derived from YB-1(-/-) embryos displayed a normal rate of protein synthesis and minimal alterations in the transcriptome and proteome but demonstrated reduced abilities to respond to oxidative, genotoxic, and oncogene-induced stresses. YB-1(-/-) cells under oxidative stress expressed high levels of the G(1)-specific CDK inhibitors p16Ink4a and p21Cip1 and senesced prematurely; this defect was corrected by knocking down CDK inhibitor levels with specific small interfering RNAs. These data suggest that YB 1 normally represses the transcription of CDK inhibitors, making it an important component of the cellular stress response signaling pathway. PMID- 15899866 TI - TAF9b (formerly TAF9L) is a bona fide TAF that has unique and overlapping roles with TAF9. AB - TFIID plays a key role in transcription initiation of RNA polymerase II preinitiation complex assembly. TFIID is comprised of the TATA box binding protein (TBP) and 14 TBP-associated factors (TAFs). A second set of transcriptional regulatory multiprotein complexes containing TAFs has been described (called SAGA, TFTC, STAGA, and PCAF/GCN5). Using matrix-assisted laser desorption ionization mass spectrometry, we identified a novel TFTC subunit, human TAF9Like, encoded by a TAF9 paralogue gene. We show that TAF9Like is a subunit of TFIID, and thus, it will be called TAF9b. TFIID and TFTC complexes in which both TAF9 and TAF9b are present exist. In vitro and in vivo experiments indicate that the interactions between TAF9b and TAF6 or TAF9 and TAF6 histone fold pairs are similar. We observed a differential induction of TAF9 and TAF9b during apoptosis that, together with their different ability to stabilize p53, points to distinct requirements for the two proteins in gene regulation. Small interfering RNA (siRNA) knockdown of TAF9 and TAF9b revealed that both genes are essential for cell viability. Gene expression analysis of cells treated with either TAF9 or TAF9b siRNAs indicates that the two proteins regulate different sets of genes with only a small overlap. Taken together, these data demonstrate that TAF9 and TAF9b share some of their functions, but more importantly, they have distinct roles in the transcriptional regulatory process. PMID- 15899867 TI - Mapping global histone methylation patterns in the coding regions of human genes. AB - Histone methylation patterns in the human genome, especially in euchromatin regions, have not been systematically characterized. In this study, we examined the profile of histone H3 methylation (Me) patterns at different lysines (Ks) in the coding regions of human genes by genome-wide location analyses by using chromatin immunoprecipitation linked to cDNA arrays. Specifically, we compared H3 KMe marks known to be associated with active gene expression, namely, H3-K4Me, H3 K36Me, and H3-K79Me, as well as those associated with gene repression, namely, H3 K9Me, H3-K27Me, and H4-K20Me. We further compared these to histone lysine acetylation (H3-K9/14Ac). Our results demonstrated that: first, close correlations are present between active histone marks except between H3-K36Me2 and H3-K4Me2. Notably, histone H3-K79Me2 is closely associated with H3-K4Me2 and H3-K36Me2 in the coding regions. Second, close correlations are present between histone marks associated with gene silencing such as H3-K9Me3, H3-K27Me2, and H4 K20Me2. Third, a poor correlation is observed between euchromatin marks (H3 K9/K14Ac, H3-K4Me2, H3-K36Me2, and H3-K79Me2) and heterochromatin marks (H3 K9Me2, H3-K9Me3, H3-K27Me2, and H4-K20Me2). Fourth, H3-K9Me2 is neither associated with active nor repressive histone methylations. Finally, histone H3 K4Me2, H3-K4Me3, H3-K36Me2, and H3-K79Me2 are associated with hyperacetylation and active genes, whereas H3-K9Me2, H3-K9Me3, H3-K27Me2, and H4-K20Me2 are associated with hypoacetylation. These data provide novel new information regarding histone KMe distribution patterns in the coding regions of human genes. PMID- 15899868 TI - Identification and characterization of a Drosophila proteasome regulatory network. AB - Maintaining adequate proteasomal proteolytic activity is essential for eukaryotic cells. For metazoan cells, little is known about the composition of genes that are regulated in the proteasome network or the mechanisms that modulate the levels of proteasome genes. Previously, two distinct treatments have been observed to induce 26S proteasome levels in Drosophila melanogaster cell lines, RNA interference (RNAi)-mediated inhibition of the 26S proteasome subunit Rpn10/S5a and suppression of proteasome activity through treatment with active site inhibitors. We have carried out genome array profiles from cells with decreased Rpn10/S5a levels using RNAi or from cells treated with proteasome inhibitor MG132 and have thereby identified candidate genes that are regulated as part of a metazoan proteasome network. The profiles reveal that the majority of genes that were identified to be under the control of the regulatory network consisted of 26S proteasome subunits. The 26S proteasome genes, including three new subunits, Ubp6p, Uch-L3, and Sem1p, were found to be up-regulated. A number of genes known to have proteasome-related functions, including Rad23, isopeptidase T, sequestosome, and the genes for the segregase complex TER94/VCP Ufd1-Npl4 were also found to be up-regulated. RNAi-mediated inhibition against the segregase complex genes demonstrated pronounced stabilization of proteasome substrates throughout the Drosophila cell. Finally, transcriptional reporter assays and deletion mapping studies in Drosophila demonstrate that proteasome mRNA induction is dependent upon the 5' untranslated regions (UTRs). Transfer of the 5' UTR from the proteasome subunit Rpn1/S2 to a noninducible promoter was sufficient to confer transcriptional upregulation of the reporter mRNA after proteasome inhibition. PMID- 15899869 TI - Graded mitogen-activated protein kinase activity precedes switch-like c-Fos induction in mammalian cells. AB - The mitogen-activated protein kinase (MAPK) pathway is an evolutionarily conserved signaling module that controls important cell fate decisions in a variety of physiological contexts. During Xenopus oocyte maturation, the MAPK cascade converts an increasing progesterone stimulus into a switch-like, all-or nothing response. While the importance of such switch-like behavior is widely discussed in the literature, it is not known whether the MAPK pathway in mammalian cells exhibits a switch-like or graded response. For this study, we used flow cytometry and immunofluorescence to generate single-cell measurements of MAPK signaling in Swiss 3T3 fibroblasts. In contrast to the case in Xenopus oocytes, we found that ERK activation in individual mammalian cells is not ultrasensitive and shows a graded response to changes in agonist concentration. Thus, the conserved MAPK signaling module exhibits different systems-level properties in different cellular contexts. Furthermore, the graded ERK response was converted into a more switch-like behavior at the level of immediate-early gene induction and cell cycle progression. Thus, while MAPK signaling is involved in all-or-nothing cell fate decisions for both Xenopus oocyte maturation and mammalian fibroblast proliferation, the underlying mechanisms responsible for the switch-like nature of the cellular responses are different in these two systems, with the mechanism appearing to lie downstream of the kinase cascade in mammalian fibroblasts. PMID- 15899870 TI - Smad1 and Smad8 function similarly in mammalian central nervous system development. AB - Smads 1, 5, and 8 are the intracellular mediators for the bone morphogenetic proteins (BMPs), which play crucial roles during mammalian development. Previous research has shown that Smad1 is important in the formation of the allantois, while Smad5 has been shown to be critical in the process of angiogenesis. To further analyze the BMP-responsive Smads, we disrupted the murine Smad8 gene utilizing the Cre/loxP system. A Smad8 hypomorphic allele (Smad8(Deltaexon3)) was constructed that contains an in-frame deletion of exon 3, removing one-third of the MH2 domain and a small portion of the linker region. Xenopus injection assays indicated that this Smad8 deletion allele is still functional but has reduced ventralizing capability compared to the wild type. Although Smad8(Deltaexon3/Deltaexon3) embryos are phenotypically normal, homozygotes of another hypomorphic allele of Smad8 (Smad8(3loxP)) containing a neomycin cassette within intron 3, phenocopy an embryonic brain defect observed in roughly 22% of Smad1(+/)(-) embryos analyzed at embryonic day 11.5. These observations suggest that BMP-responsive Smads have critical functions in the development of the mammalian central nervous system. PMID- 15899871 TI - Selective role of a distinct tyrosine residue on Tie2 in heart development and early hematopoiesis. AB - The development of the cardiovascular system and the development of the early hematopoietic systems are closely related, and both require signaling through the Tie2 receptor tyrosine kinase. Although endothelial cells and hematopoietic cells as well as their precursors share common gene expression patterns during development, it remains completely unknown how Tie2 signaling coordinately regulates cardiovascular development and early hematopoiesis in vivo. We show here that mice with a targeted mutation in tyrosine residue 1100 in the carboxyl terminal tail of Tie2 display defective cardiac development and impaired hematopoietic and endothelial cell development in the paraaortic splanchnopleural mesoderm similar to that seen in Tie2-null mutant mice. Surprisingly, however, unlike Tie2-null mutant mice, mice deficient in signaling through this tyrosine residue show a normal association of perivascular cells with nascent blood vessels. These studies are the first to demonstrate the physiological importance of a single tyrosine residue in Tie2, and they suggest that multiple tyrosine residues in the receptor may coordinate cardiovascular development and early hematopoietic development. PMID- 15899872 TI - Transforming growth factor {beta} (TGF-{beta})-Smad target gene protein tyrosine phosphatase receptor type kappa is required for TGF-{beta} function. AB - Transforming growth factor beta (TGF-beta) inhibits proliferation and promotes cell migration. In TGF-beta-treated MCF10A mammary epithelial cells overexpressing HER2 and by chromatin immunoprecipitation, we identified novel Smad targets including protein tyrosine phosphatase receptor type kappa (PTPRK). TGF-beta up-regulated PTPRK mRNA and RPTPkappa (receptor type protein tyrosine phosphatase kappa, the protein product encoded by the PTPRK gene) protein in tumor and nontumor mammary cells; HER2 overexpression down-regulated its expression. RNA interference (RNAi) of PTPRK accelerated cell cycle progression, enhanced response to epidermal growth factor (EGF), and abrogated TGF-beta mediated antimitogenesis. Endogenous RPTPkappa associated with EGF receptor and HER2, resulting in suppression of basal and ErbB ligand-induced proliferation and receptor phosphorylation. In MCF10A/HER2 cells, TGF-beta enhanced cell motility, FAK phosphorylation, F-actin assembly, and focal adhesion formation and inhibited RhoA activity. These responses were abolished when RPTPkappa was eliminated by RNA interference (RNAi). In cells expressing RPTPkappa RNAi, phosphorylation of Src at Tyr527 was increased and (activating) phosphorylation of Src at Tyr416 was reduced. These data suggest that (i) RPTPkappa positively regulates Src; (ii) HER2 signaling and TGF-beta-induced RPTPkappa converge at Src, providing an adequate input for activation of FAK and increased cell motility and adhesion; and (iii) RPTPkappa is required for both the antiproliferative and the promigratory effects of TGF-beta. PMID- 15899873 TI - Ankyrin repeat and SOCS box 3 (ASB3) mediates ubiquitination and degradation of tumor necrosis factor receptor II. AB - Ankyrin repeat and SOCS box (ASB) family members have a C-terminal SOCS box and an N-terminal ankyrin-related sequence of variable repeats belonging to the SOCS superfamily. While SH2-domain-bearing SOCS proteins are mainly involved in the negative feedback regulation of the protein tyrosine kinase-STAT pathway in response to a variety of cytokines, the roles of ASB family members remain largely unknown. To investigate ASB functions, we screened for ASB3-interacting factors by using antibody array technology and identified tumor necrosis factor receptor II (TNF-R2) as an ASB3 binding target. ASB3 expression and activities are required for (i) TNF-R2 ubiquitination both in vivo and in vitro, (ii) TNF-R2 proteolysis via the proteasome pathway, and (iii) the inhibition of TNF-R2 mediated Jun N-terminal protein kinase (JNK) activation. While the ankyrin repeats of ASB3 interact with the C-terminal 37 amino acids of TNF-R2, the SOCS box of ASB3 is responsible for recruiting the E3 ubiquitin ligase adaptors Elongins-B/C, leading to TNF-R2 ubiquitination on multiple lysine residues within its C-terminal region. Downregulation of ASB3 expression by a small interfering RNA inhibited TNF-R2 degradation and potentiated TNF-R2-mediated cytotoxicity. The data presented here implicate ASB3 as a negative regulator of TNF-R2-mediated cellular responses to TNF-alpha by direct targeting of TNF-R2 for ubiquitination and proteasome-mediated degradation. PMID- 15899874 TI - 5-Aza-deoxycytidine induces selective degradation of DNA methyltransferase 1 by a proteasomal pathway that requires the KEN box, bromo-adjacent homology domain, and nuclear localization signal. AB - 5-Azacytidine- and 5-aza-deoxycytidine (5-aza-CdR)-mediated reactivation of tumor suppressor genes silenced by promoter methylation has provided an alternate approach in cancer therapy. Despite the importance of epigenetic therapy, the mechanism of action of DNA-hypomethylating agents in vivo has not been completely elucidated. Here we report that among three functional DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B), the maintenance methyltransferase, DNMT1, was rapidly degraded by the proteasomal pathway upon treatment of cells with these drugs. The 5-aza-CdR-induced degradation, which occurs in the nucleus, could be blocked by proteasomal inhibitors and required a functional ubiquitin-activating enzyme. The drug-induced degradation occurred even in the absence of DNA replication. Treatment of cells with other nucleoside analogs modified at C-5, 5 fluorodeoxyuridine and 5-fluorocytidine, did not induce the degradation of DNMT1. Mutation of cysteine at the catalytic site of Dnmt1 (involved in the formation of a covalent intermediate with cytidine in DNA) to serine (CS) did not impede 5-aza CdR-induced degradation. Neither the wild type nor the catalytic site mutant of Dnmt3a or Dnmt3b was sensitive to 5-aza-CdR-mediated degradation. These results indicate that covalent bond formation between the enzyme and 5-aza-CdR incorporated DNA is not essential for enzyme degradation. Mutation of the conserved KEN box, a targeting signal for proteasomal degradation, to AAA increased the basal level of Dnmt1 and blocked its degradation by 5-aza-CdR. Deletion of the catalytic domain increased the expression of Dnmt1 but did not confer resistance to 5-aza-CdR-induced degradation. Both the nuclear localization signal and the bromo-adjacent homology domain were essential for nuclear localization and for the 5-aza-CdR-mediated degradation of Dnmt1. Polyubiquitination of Dnmt1 in vivo and its stabilization upon treatment of cells with a proteasomal inhibitor indicate that the level of Dnmt1 is controlled by ubiquitin-dependent proteasomal degradation. Overexpression of the substrate recognition component, Cdh1 but not Cdc20, of APC (anaphase-promoting complex)/cyclosome ubiquitin ligase reduced the level of Dnmt1 in both untreated and 5-aza-CdR-treated cells. In contrast, the depletion of Cdh1 with small interfering RNA increased the basal level of DNMT1 that blocked 5-aza-CdR-induced degradation. Dnmt1 interacted with Cdh1 and colocalized in the nucleus at discrete foci. Both Dnmt1 and Cdh1 were phosphorylated in vivo, but only Cdh1 was significantly dephosphorylated upon 5-aza-CdR treatment, suggesting its involvement in initiating the proteasomal degradation of DNMT1. These results demonstrate a unique mechanism for the selective degradation of DNMT1, the maintenance DNA methyltransferase, by well-known DNA-hypomethylating agents. PMID- 15899875 TI - FLIP protects against hypoxia/reoxygenation-induced endothelial cell apoptosis by inhibiting Bax activation. AB - Hypoxia/reoxygenation causes cell death, yet the underlying regulatory mechanisms remain partially understood. Recent studies demonstrate that hypoxia/reoxygenation can activate death receptor and mitochondria-dependent apoptotic pathways, involving Bid and Bax mitochondrial translocation and cytochrome c release. Using mouse lung endothelial cells (MLEC), we examined the role of FLIP, an inhibitor of caspase 8, in hypoxia/reoxygenation-induced cell death. FLIP protected MLEC against hypoxia/reoxygenation by blocking both caspase 8/Bid and Bax/mitochondrial apoptotic pathways. FLIP inhibited Bax activation in wild-type and Bid(-/-) MLEC, indicating independence from the caspase 8/Bid pathway. FLIP also inhibited the expression and activation of protein kinase C (PKC) (alpha, zeta) during hypoxia/reoxygenation and promoted an association of inactive forms of PKC with Bax. Surprisingly, FLIP expression also inhibited death-inducing signal complex (DISC) formation in the plasma membrane and promoted the accumulation of the DISC in the Golgi apparatus. FLIP expression also upregulated Bcl-X(L), an antiapoptotic protein. In conclusion, FLIP decreased DISC formation in the plasma membrane by blocking its translocation from the Golgi apparatus and inhibited Bax activation through a novel PKC dependent mechanism. The inhibitory effects of FLIP on Bax activation and plasma membrane DISC formation may play significant roles in protecting endothelial cells from the lethal effects of hypoxia/reoxygenation. PMID- 15899876 TI - Identification of a conserved RNA motif essential for She2p recognition and mRNA localization to the yeast bud. AB - In Saccharomyces cerevisiae, over twenty mRNAs localize to the bud tip of daughter cells, playing roles in processes as different as mating type switching and plasma membrane targeting. The localization of these transcripts depends on interactions between a cis-acting localization element(s) or zipcodes and the RNA binding protein She2p. While previous studies identified four different localization elements in the bud-localized ASH1 mRNA, the main determinants for She2p recognition are still unknown. To investigate the RNA-binding specificity of She2p, we isolated She2p-binding RNAs by in vivo selection from libraries of partially randomized ASH1 localization elements. The RNAs isolated contained a similar loop-stem-loop structure with a highly conserved CGA triplet in one loop and a single conserved cytosine in the other loop. Mutating these conserved nucleotides or the stem separating them resulted in the loss of She2p binding and in the delocalization of a reporter mRNA. Using this information, we identified the same RNA motif in two other known bud-localized transcripts, suggesting that this motif is conserved among bud-localized mRNAs. These results show that mRNAs with zipcodes lacking primary sequence similarity can rely on a few conserved nucleotides properly oriented in their three-dimensional structure in order to be recognized by the same localization machinery. PMID- 15899877 TI - Novel response to microtubule perturbation in meiosis. AB - During the mitotic cell cycle, microtubule depolymerization leads to a cell cycle arrest in metaphase, due to activation of the spindle checkpoint. Here, we show that under microtubule-destabilizing conditions, such as low temperature or the presence of the spindle-depolymerizing drug benomyl, meiotic budding yeast cells arrest in G(1) or G(2), instead of metaphase. Cells arrest in G(1) if microtubule perturbation occurs as they enter the meiotic cell cycle and in G(2) if cells are already undergoing premeiotic S phase. Concomitantly, cells down-regulate genes required for cell cycle progression, meiotic differentiation, and spore formation in a highly coordinated manner. Decreased expression of these genes is likely to be responsible for halting both cell cycle progression and meiotic development. Our results point towards the existence of a novel surveillance mechanism of microtubule integrity that may be particularly important during specialized cell cycles when coordination of cell cycle progression with a developmental program is necessary. PMID- 15899878 TI - DNA methylation precedes chromatin modifications under the influence of the strain-specific modifier Ssm1. AB - Ssm1 is responsible for the mouse strain-specific DNA methylation of the transgene HRD. In adult mice of the C57BL/6 (B6) strain, the transgene is methylated at essentially all CpGs. However, when the transgene is bred into the DBA/2 (D2) strain, it is almost completely unmethylated. Strain-specific methylation arises during differentiation of embryonic stem (ES) cells. Here we show that Ssm1 causes striking chromatin changes during the development of the early embryo in both strains. In undifferentiated ES cells of both strains, the transgene is in a chromatin state between active and inactive. These states are still observed 1 week after beginning ES cell differentiation. However, 4 weeks after initiating differentiation, in B6, the transgene has become heterochromatic, and in D2, the transgene has become euchromatic. HRD is always expressed in D2, but in B6, it is expressed only in early embryos. The transgene is already more methylated in B6 ES cells than in D2 ES cells and becomes increasingly methylated during development in B6, until essentially all CpGs in the critical guanosine phosphoribosyl transferase core are methylated. Clearly, DNA methylation of HRD precedes chromatin compaction and loss of expression, suggesting that the B6 form of Ssm1 interacts with DNA to cause strain-specific methylation that ultimately results in inactive chromatin. PMID- 15899879 TI - The noncatalytic amino terminus of mitogen-activated protein kinase phosphatase 1 directs nuclear targeting and serum response element transcriptional regulation. AB - The mitogen-activated protein kinase (MAPK) phosphatase 1 (MKP-1) is an immediate early gene comprised of a dual-specificity phosphatase domain and a noncatalytic NH(2) terminus. Here, we show that the NH(2) terminus of MKP-1, containing the cdc25 homology domains A (CH2A) and B (CH2B), mediates MKP-1 nuclear targeting and modulates MAPK-mediated gene expression. An LXXLL motif which is known to mediate protein-protein interactions with nuclear-targeted hormone receptors was identified proximal to the CH2A domain of MKP-1. The NH(2) terminus alone of MKP 1 containing this LXXLL motif was sufficient to direct nuclear targeting, and mutating this motif to LXXAA resulted in the exclusion of MKP-1 from the nucleus. We found that the LXXLL motif proximal to the CH2A domain was present in other nuclear-localized MKPs but was absent in MKPs that localized to the cytoplasm. These data suggest that this LXXLL motif confers nuclear targeting properties to the MKPs. The NH(2) terminus of MKP-1 was also found to inhibit the activation of the serum response element (SRE) by preventing MAPK-mediated phosphorylation of the regulatory serine 383 residue on Elk-1. Moreover, we show that MKP-1 plays a major role in the attenuation of serum-induced SRE activity, since MKP-1 null fibroblasts exhibited enhanced SRE activity in response to serum compared with wild-type fibroblasts. The NH(2) terminus of MKP-1, when reconstituted into MKP-1 null fibroblasts to levels similar to endogenous MKP-1 following serum stimulation, reduced serum-mediated SRE activity. Collectively, these data reveal novel roles for the NH(2) terminus of MKP-1 in nuclear targeting and transcriptional regulation. PMID- 15899880 TI - Phenotypic analysis of mice deficient in the type 2 galanin receptor (GALR2). AB - Galanin is a neuropeptide implicated in the regulation of feeding, reproduction, cognition, nociception, and seizure susceptibility. There are three known galanin receptor (GALR) subtypes (GALR1, GALR2, and GALR3), which bind to galanin with different affinities and have their own unique distributions, signaling mechanisms, and putative functions in the brain and peripheral nervous system. To gain further insight into the possible physiological significance of GALR2, we created mutant mice that were deficient in GALR2 and compared their phenotype to that of wild-type (WT) littermate or age-matched controls, with respect to basic motor and sensory function, feeding behavior, reproduction, mood, learning and memory, and seizure susceptibility. Phenotypic analysis revealed that animals bearing a deletion of GALR2 did not differ significantly from their WT controls in any of the measured variables. We conclude that either GALR2 plays no role in these physiological functions or through redundancy or compensation these mutant animals can adapt to the congenital absence of GALR2. It is also conceivable that GALR2 plays only a subtle role in some of these functions and that the impact of its loss could not be detected by the analytical procedures used here. PMID- 15899881 TI - Promoter trapping in Lotus japonicus reveals novel root and nodule GUS expression domains. AB - Agrobacterium-based transformation was used to introduce a promoter-less glucuronidase uidA gene (beta-glucuronidase; GUS) into Lotus japonicus. Transgenic plants were screened for GUS activation at different stages after inoculation with its symbiont, Mesorhizobium loti. Functional GUS fusion frequencies ranged from about 2 to 5% of the total number of transgenic lines. These lines provide excellent histological markers for tissue ontogeny analysis. Some of the activations generated GUS expression patterns that correspond to well known tissue types, such as lateral root and nodule primordia, root tips and developing nodules (line CHEETAH). Others generated GUS activation associated with predictable but previously unknown (i) tissue types, such as the vascular bundle of the nodule (line VASCO); or (ii) expression domains, such as pericycle, nodule primordia, nodule and flower connective/vascular tissue (line FATA MORGANA) or inner root cortex cells in the vicinity of a curled root hair, nodule primordia and nodule cortex (line TIMPA). Putative members of two gene superfamilies, EH (Esp homolog) and AAA ATPase (ATPase associated with various cellular activities), were located next to the CHEETAH and VASCO insertions, respectively, and a nodulin gene, LjENOD40-2, was located next to the FATA MORGANA insertion. We utilized promoter GUS fusions to investigate the genetic regulation of LjENOD40-2 and FATA MORGANA GUS. The LjENOD40-2 promoter defined a novel expression domain and the FATA MORGANA nodule expression was reiterated by the 2 kb sequence upstream of the T-DNA insertion. PMID- 15899883 TI - Role for protein phosphatase 2A in the regulation of Calu-3 epithelial Na+-K+-2Cl , type 1 co-transport function. AB - Activity of Na+-K+-2Cl- co-transport (NKCC1) in epithelia is thought to be highly regulated through phosphorylation and dephosphorylation of the transporter. Previous functional studies from this laboratory suggested a role for protein phosphatase 2A (PP2A) as a serine/threonine protein phosphatase involved in the regulation of mammalian tracheal epithelial NKCC1. We expand on these studies to characterize serine/threonine protein phosphatase(s) necessary for regulation of NKCC1 function and the interaction of the phosphatase(s) with proteins associated with NKCC1. NKCC1 activity was measured as bumetanide-sensitive 86Rb uptake or basolateral to apical 86Rb flux in primary cultures of human tracheal epithelial cells or in Calu-3 airway epithelial cells grown on Transwell filter inserts. Preincubation with 0.1 nm okadaic acid, a PP2A >> phosphatase 1 (PP1) inhibitor, increased NKCC1 activity 3.5-fold in human tracheal epithelial cells and 4.1-fold in Calu-3 cells. Calyculin, a PP1 >> PP2A inhibitor, did not alter NKCC1 activity or percent bumetanide-sensitive flux. The effect of OA was dose-dependent with an IC50 of 0.4 nm. The alpha1-adrenergic agonist methoxamine increased NKCC1 activity and transiently increased PP2A activity 3.8-fold but did not alter PP1 activity. OA augmented methoxamine-dependent stimulation of NKCC1 activity. PP1, PP2A, and PP2C but not PP2B were detected in lysates from Calu-3 cells by immunoblot analysis. PP1 was not detected in immunoprecipitates of NKCC1 and vice versa. PP2A co-immunoprecipitated with NKCC1 and protein kinase C-delta (PKC delta) and was pulled down by a recombinant N terminus of NKCC1 consisting of amino acids 1-286. One novel finding is co-precipitation of STE20-related proline alanine-rich kinase, a regulatory kinase for NKCC1, with PP2A and PKC-delta. The results suggest a model of actin serving as a scaffold for binding and association of PKC-delta, PP2A, and STE20-related proline-alanine-rich kinase. The role of the complex of serine/threonine protein kinases and a protein phosphatase is probably the maintenance of optimal phosphorylation of NKCC1 coincident with its physiological function in epithelial absorption and secretion. PMID- 15899882 TI - An intersubunit zinc binding site in rat P2X2 receptors. AB - P2X receptors are ATP-gated ion channels made up of three similar or identical subunits. It is unknown whether ligand binding is intersubunit or intrasubunit, either for agonists or for allosteric modulators. Zinc binds to rat P2X2 receptors and acts as an allosteric modulator, potentiating channel opening. To probe the location of this zinc binding site, P2X2 receptors bearing mutations of the histidines at positions 120 and 213 were expressed in Xenopus oocytes. Studies of H120C and H213C mutants produced five lines of evidence consistent with the hypothesis that the residues in these positions bind zinc. Mixing of subunits containing the H120A or H213A mutation generated receptors that showed zinc potentiation, even though neither of these mutant receptors showed zinc potentiation on its own. Furthermore, expression of trimeric concatamers with His --> Ala mutations at some but not all six positions showed that zinc potentiation correlated with the number of intersubunit histidine pairs. These results indicate that zinc potentiation requires an interaction across a subunit interface. Expression of the H120C/H213C double mutant resulted in the formation of ectopic disulfide bonds that could be detected by changes in the physiological properties of the receptors after treatment with reducing and oxidizing agents. Immunoblot analysis of H120C/H213C protein separated under nonreducing conditions demonstrated that the ectopic bonds were between adjacent subunits. Taken together, these data indicate that His120 and His213 sit close to each other across the interface between subunits and are likely to be key components of the zinc binding site in P2X2 receptors. PMID- 15899884 TI - Differential regulation of amino acid transporter SNAT3 by insulin in hepatocytes. AB - The liver is a metabolism and transfer center of amino acids as well as the prime target organ of insulin. In this report, we characterized the regulation of system N/A transporter 3 (SNAT3) in the liver of dietary-restricted mice and in hepatocytes treated with serum starvation and insulin. The expression of SNAT3 was up-regulated in dietary-restricted mice. The expression of SNAT3 protein was detected on the plasma membrane of hepatocyte-like H2.35 cells with a half-life of 6-8 h. When H2.35 cells were depleted of serum, the expression of SNAT3 was increased. An increased concentration of insulin, however, suppressed SNAT3 expression. Interestingly, the down-regulation of SNAT3 expression by insulin was blocked by the specific phosphoinositide 3-kinase inhibitor LY294002 and mammalian target of rapamycin inhibitor, but not by MAPK inhibitor PD98059, suggesting that insulin exerts its effect on SNAT3 through phosphoinositide 3 kinase-mammalian target of rapamycin signaling. Surface biotinylation assay showed an increased level of SNAT3 on the cell surface after 0.5 h of insulin treatment, although no effect was observed after 24 h of treatment. Consistently, the transport of the substrate l-histidine was increased with short, but not long, treatment by insulin in both H2.35- and SNAT3-transfected COS-7 cells. The L-histidine uptake was inhibited significantly by L-histidine followed by 2 endoamino-bicycloheptane-2-carboxylic acid and L-cysteine and to a lesser extent by L-alanine and aminoisobutyric acid, but was not inhibited by alpha (methylamino)isobutyric acid, implying that uptake of L-histidine in H2.35 cells is primarily mediated by system N transporters. In conclusion, differential regulation of SNAT3 by insulin and serum starvation reinforces the functional significance of this transporter in liver physiology. PMID- 15899885 TI - Insig required for sterol-mediated inhibition of Scap/SREBP binding to COPII proteins in vitro. AB - When added to living cells, sterols such as cholesterol and 25-hydroxycholesterol block the lateral movement of sterol regulatory element-binding proteins (SREBPs) into COPII-coated vesicles on endoplasmic reticulum (ER) membranes and thereby prevent the SREBPs from reaching the Golgi complex for processing to the mature forms that activate cholesterol synthesis. Sorting of SREBPs into COPII vesicles is mediated by Sar1 and the coat proteins Sec23 and Sec24. Here, we explore the mechanism of sterol inhibition in vitro through use of protein pull-down assays. We show that addition of cholesterol or 25-hydroxycholesterol to microsomal membranes in vitro blocks Sar1-dependent binding of the Sec23/24 complex to Scap, the SREBP escort protein. This in vitro inhibition is dependent on the presence of Insig-1, an ER resident protein that is necessary for sterol-mediated inhibition of Scap/SREBP transport in intact cells. Sec23/24 binding to Scap requires the hexapeptide sequence MELADL located in a cytoplasmic loop of Scap. This hexapeptide acts as a sterol-regulated ER sorting signal. These studies define the biochemical parameters responsible for regulated sorting of an ER membrane protein into COPII-coated vesicles. PMID- 15899886 TI - ERK1/2-dependent activation of transcription factors required for acute and chronic effects of glucose on the insulin gene promoter. AB - The insulin promoter is both positively and negatively regulated in response to conditions to which pancreatic beta-cells are exposed. Exposure of intact rat islets and INS-1 pancreatic beta-cells to 11 mm glucose for minutes to hours results in an enhancement in the rate of insulin gene transcription assessed with a reporter linked to the insulin gene promoter. In contrast, chronic exposure of rat islets or beta-cells to 11 mm glucose results in loss of the glucose responsiveness of the insulin gene promoter. By 48 h, glucose inhibits insulin gene promoter activity. Here we show that not only the acute effect of elevated glucose to stimulate the insulin gene promoter but also the chronic effect of elevated glucose to inhibit the insulin gene promoter depend on ERK1/2 mitogen activated protein kinase activity. In examining the underlying mechanism, we found that acute exposure to 11 mm glucose resulted in the binding of the transcription factors NFAT and Maf to the glucose-responsive A2C1 element of the insulin gene promoter. An NFAT and C/EBP-beta complex was observed in cells chronically exposed to 11 mm glucose. Formation of NFAT-Maf and NFAT-C/EBP-beta complexes was sensitive to inhibitors of ERK1/2 and calcineurin, consistent with our previous finding that activation of ERK1/2 by glucose required calcineurin activity and the well documented regulation of NFAT by calcineurin. These results indicate that the ERK1/2 pathway modulates partners of NFAT, which may either stimulate or repress insulin gene transcription during stimulatory and chronic exposure to elevated glucose. PMID- 15899887 TI - Novel autoregulatory function of hepatitis B virus M protein on surface gene expression. AB - The hepatitis B virus surface gene consists of a single open reading frame divided into three coding regions: pre-S1, pre-S2, and S. By alternate translation at each of the three initiation codons, L, M, and S proteins can be synthesized. Studies have shown that M protein is not essential for viral replication, virion morphogenesis, or in vitro infectivity. In this study, we show that native M protein can regulate surface gene expression at the transcriptional level. The regulatory effect of M protein is mediated through the CCAAT box within the S promoter. Deletion mapping analysis indicated that the transactivating effect of M protein is mediated through amino acids 1-57 of M protein (the MHBs(au) domain), although its maximal transactivation activity coincides with that of the pre-S2 domain. This conclusion is supported by the fact that disruption of the putative V8 protease site at the pre-S2/S domain junction not only rendered M protein incapable of transactivating the S promoter but also inactivated its nuclear translocation potential. Immunoprecipitation and immunoblot experiments demonstrated that pre-S2 interacts with the three subunits of the CCAAT box-binding factor/nuclear factor Y, the cognate binding protein of the CCAAT box. These results demonstrate and define a novel regulatory role of M protein, which, under natural conditions, may undergo a proteolytic process to generate an MHBs(au) species that will be translocated inside the nucleus, where it will interact with the CCAAT box-binding factor to regulate surface gene expression. Because the CCAAT box is located at a fixed position within numerous promoters, these observations might provide a plausible explanation for hepatitis B virus-associated hepatocarcinogenesis. PMID- 15899888 TI - The farnesoid X receptor modulates hepatic carbohydrate metabolism during the fasting-refeeding transition. AB - The liver plays a central role in the control of blood glucose homeostasis by maintaining a balance between glucose production and utilization. The farnesoid X receptor (FXR) is a bile acid-activated nuclear receptor. Hepatic FXR expression is regulated by glucose and insulin. Here we identify a role for FXR in the control of hepatic carbohydrate metabolism. When submitted to a controlled fasting-refeeding schedule, FXR(-/-) mice displayed an accelerated response to high carbohydrate refeeding with an accelerated induction of glycolytic and lipogenic genes and a more pronounced repression of gluconeogenic genes. Plasma insulin and glucose levels were lower in FXR(-/-) mice upon refeeding the high carbohydrate diet. These alterations were paralleled by decreased hepatic glycogen content. Hepatic insulin sensitivity was unchanged in FXR(-/-) mice. Treatment of isolated primary hepatocytes with a synthetic FXR agonist attenuated glucose-induced mRNA expression as well as promoter activity of L-type pyruvate kinase, acetyl-CoA carboxylase 1, and Spot14. Moreover, activated FXR interfered negatively with the carbohydrate response elements regions. These results identify a novel role for FXR as a modulator of hepatic carbohydrate metabolism. PMID- 15899889 TI - Phosphorylation of mammalian target of rapamycin (mTOR) at Ser-2448 is mediated by p70S6 kinase. AB - The mammalian target of rapamycin (mTOR) coordinates cell growth with the growth factor and nutrient/energy status of the cell. The phosphatidylinositol 3-kinase AKT pathway is centrally involved in the transmission of mitogenic signals to mTOR. Previous studies have shown that mTOR is a direct substrate for the AKT kinase and identified Ser-2448 as the AKT target site in mTOR. In this study, we demonstrate that rapamycin, a specific inhibitor of mTOR function, blocks serum stimulated Ser-2448 phosphorylation and that this drug effect is not explained by the inhibition of AKT. Furthermore, the phosphorylation of Ser-2448 was dependent on mTOR kinase activity, suggesting that mTOR itself or a protein kinase downstream from mTOR was responsible for the modification of Ser-2448. Here we show that p70S6 kinase phosphorylates mTOR at Ser-2448 in vitro and that ectopic expression of rapamycin-resistant p70S6 kinase restores Ser-2448 phosphorylation in rapamycin-treated cells. In addition, we show that cellular amino acid status, which modulates p70S6 kinase (S6K1) activity via the TSC/Rheb pathway, regulates Ser-2448 phosphorylation. Finally, small interfering RNA-mediated depletion of p70S6 kinase reduces Ser-2448 phosphorylation in cells. Taken together, these results suggest that p70S6 kinase is a major effector of mTOR phosphorylation at Ser-2448 in response to both mitogen- and nutrient-derived stimuli. PMID- 15899890 TI - Janus kinases affect thrombopoietin receptor cell surface localization and stability. AB - The thrombopoietin receptor (TpoR) regulates hematopoietic stem cell renewal, megakaryocyte differentiation, and platelet formation. TpoR signals by activating Janus kinases JAK2 and Tyk2. Here we show that, in addition to signaling downstream from the activated TpoR, JAK2 and Tyk2 strongly promote cell surface localization and enhance total protein levels of the TpoR. This effect is caused by stabilization of the mature endoglycosidase H-resistant form of the receptor. Confocal microscopy indicates that TpoR colocalizes partially with recycling transferrin in Ba/F3 cells. The interaction with JAK2 or Tyk2 appears to protect the receptor from proteasome degradation. Sequences encompassing Box1 and Box2 regions of the receptor cytosolic domain and an intact JAK2 or Tyk2 FERM domain are required for these effects. We discuss the relevance of our results to the reported defects of TpoR processing in myeloproliferative diseases and to the mechanisms of Tpo signaling and clearance via the TpoR. PMID- 15899891 TI - Ceramide 1-phosphate, a mediator of phagocytosis. AB - The agonist-stimulated metabolism of membrane lipids produces potent second messengers that regulate phagocytosis. We studied whether human ceramide kinase (hCERK) activity and ceramide 1-phosphate formation could lead to enhanced phagocytosis through a mechanism involving modulation of the membrane-structural order parameter. hCERK was stably transfected into COS-1 cells that were stably transfected with the FcgammaRIIA receptor. hCERK-transfected cells displayed a significant increase in phagocytic index in association with increased ceramide kinase activation and translocation to lipid rafts after activation with opsonized erythrocytes. When challenged with opsonized erythrocytes, hCERK transfected cells increased phagocytosis by 1.5-fold compared with vector control and simultaneously increased ceramide 1-phosphate levels 2-fold compared with vector and unstimulated control cells. Control and hCERK-transfected cells were subjected to cellular fractionation. Utilizing an antibody against hCERK, we observed that CERK translocates during activation from the cytosol to a lipid raft fraction. The plasma membrane-structural order parameter of the transfectants was measured by labeling cells with Laurdan. Cells transfected with hCERK showed a higher liquid crystalline order than control cells with stimulation, conditions that are favorable for the promotion of membrane fusion at the sites of phagocytosis. The change in the structural order parameter of the lipid rafts probably contributes to phagocytosis by promoting phagosome formation. PMID- 15899892 TI - Biochemical analysis of the DNA unwinding and strand annealing activities catalyzed by human RECQ1. AB - RecQ helicases play an important role in preserving genomic integrity, and their cellular roles in DNA repair, recombination, and replication have been of considerable interest. Of the five human RecQ helicases identified, three are associated with genetic disorders characterized by an elevated incidence of cancer or premature aging: Werner syndrome, Bloom syndrome, and Rothmund-Thomson syndrome. Although the biochemical properties and protein interactions of the WRN and BLM helicases defective in Werner syndrome and Bloom syndrome, respectively, have been extensively investigated, less information is available concerning the functions of the other human RecQ helicases. We have focused our attention on human RECQ1, a DNA helicase whose cellular functions remain largely uncharacterized. In this work, we have characterized the DNA substrate specificity and optimal cofactor requirements for efficient RECQ1-catalyzed DNA unwinding and determined that RECQ1 has certain properties that are distinct from those of other RecQ helicases. RECQ1 stably bound to a variety of DNA structures, enabling it to unwind a diverse set of DNA substrates. In addition to its DNA binding and helicase activities, RECQ1 catalyzed efficient strand annealing between complementary single-stranded DNA molecules. The ability of RECQ1 to promote strand annealing was modulated by ATP binding, which induced a conformational change in the protein. The enzymatic properties of the RECQ1 helicase and strand annealing activities are discussed in the context of proposed cellular DNA metabolic pathways that are important in the maintenance of genomic stability. PMID- 15899893 TI - Crystal structure of acetylcholine-binding protein from Bulinus truncatus reveals the conserved structural scaffold and sites of variation in nicotinic acetylcholine receptors. AB - The crystal structure of acetylcholine-binding protein (AChBP) from the mollusk Lymnaea stagnalis is the established model for the ligand binding domains of the ligand-gated ion channel family, which includes nicotinic acetylcholine, 5 hydroxytryptamine (5-HT3), gamma-aminobutyric acid (GABA), types A and C, and glycine receptors. Here we present the crystal structure of a remote homolog, AChBP from Bulinus truncatus, which reveals both the conserved structural scaffold and the sites of variation in this receptor family. These include rigid body movements of loops that are close to the transmembrane interface in the receptors and changes in the intermonomer contacts, which alter the pentamer stability drastically. Structural, pharmacological and mutational analysis of both AChBPs shows how 3 amino acid changes in the binding site contribute to a 5 10-fold difference in affinity for nicotinic ligands. Comparison of these structures will be valuable for improving structure-function studies of ligand gated ion channel receptors, including signal transduction, homology modeling, and drug design. PMID- 15899894 TI - Estrogen inhibits cardiomyocyte hypertrophy in vitro. Antagonism of calcineurin related hypertrophy through induction of MCIP1. AB - Evidence from in vivo studies suggests that some inputs to cardiac hypertrophy are opposed by the actions of estrogen. However, the mechanisms of E2 action in this respect are mainly unknown. An important pathway that is utilized by multiple hypertrophic stimuli involves the activation of the tyrosine phosphatase, calcineurin (PP2B). Here we show that 17beta-estradiol (E2) significantly prevents angiotensin II (AngII)- or endothelin-1 (ET-1)-induced new protein synthesis, skeletal muscle actin expression, and increased surface area in cultured rat cardiomyocytes. ET-1 stimulated calcineurin phosphatase activity, resulting in new protein synthesis, and both were prevented by E2. E2 induced the MCIP1 gene, an inhibitor of calcineurin activity, via phosphatidylinositol 3 kinase, transcriptional, and mRNA stability mechanisms. Small interfering RNA for MCIP1 significantly reversed both the E2 restraint of protein synthesis and the inhibition of AngII-induced calcineurin activity. AngII-induced the translocation of the hypertrophic transcription factor, NF-AT, to the nucleus of the cardiomyocyte and stimulated NF-AT transcriptional activity. Both were prevented by E2. AngII also stimulated the activation of ERK and protein kinase C, contributing to cardiac hypertrophy. E2 inhibited these pathways, related to the stimulation of atrial natriuretic peptide production and secretion. Thus, restraint of calcineurin and kinase signaling to the hypertrophic program underlie these important effects of E2. PMID- 15899895 TI - Substrate-dependent differences in U2AF requirement for splicing in adenovirus infected cell extracts. AB - U2AF has been characterized as an essential splicing factor required for efficient recruitment of U2 small nuclear ribonucleoprotein to the 3'-splice site in a pre-mRNA. The U2AF65 subunit binds to the pyrimidine tract of the pre-mRNA, whereas the U2AF(35) subunit contacts the 3'-splice site AG. Here we show that U2AF35 appears to be completely dispensable for splicing in nuclear extracts prepared from adenovirus late-infected cells (Ad-NE). As a consequence, the viral IIIa and cellular IgM introns, which both have suboptimal 3'-splice sites and require U2AF35 for splicing in nuclear extracts from uninfected cells, are transformed to U2AF35-independent introns in Ad-NE. Furthermore, we present evidence that two parallel pathways of 3'-splice site recognition exist in Ad-NE. We show that the viral 52,55K intron, which has an extended pyrimidine tract, requires U2AF for activity in Ad-NE. In contrast, the IgM intron, which has a weak 3'-splice site sequence context, undergoes the first catalytic step of splicing in U2AF-depleted Ad-NE, suggesting that spliceosome assembly occurs through a novel U2AF-independent pathway in Ad-NE. PMID- 15899896 TI - Cannabinoids and ghrelin have both central and peripheral metabolic and cardiac effects via AMP-activated protein kinase. AB - Endocannabinoids and ghrelin are potent appetite stimulators and are known to interact at a hypothalamic level. However, both also have important peripheral actions, including beneficial effects on the ischemic heart and increasing adipose tissue deposition, while ghrelin has direct effects on carbohydrate metabolism. The AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme that functions as a fuel sensor to regulate energy balance at both cellular and whole body levels, and it may mediate the action of anti-diabetic drugs such as metformin and peroxisome proliferator-activated receptor gamma agonists. Here we show that both cannabinoids and ghrelin stimulate AMPK activity in the hypothalamus and the heart, while inhibiting AMPK in liver and adipose tissue. These novel effects of cannabinoids on AMPK provide a mechanism for a number of their known actions, such as the reduction in infarct size in the myocardium, an increase in adipose tissue, and stimulation of appetite. The beneficial effects of ghrelin on heart function, including reduction of myocyte apoptosis, and its effects on lipogenesis and carbohydrate metabolism, can also be explained by its ability to activate AMPK. Our data demonstrate that AMPK not only links the orexigenic effects of endocannabinoids and ghrelin in the hypothalamus but also their effects on the metabolism of peripheral tissues. PMID- 15899897 TI - Residue Leu-641 of Acetyl-CoA synthetase is critical for the acetylation of residue Lys-609 by the Protein acetyltransferase enzyme of Salmonella enterica. AB - Posttranslational regulation of protein function by acetylation is present throughout nature. Regulation of protein function by Sir2 protein (sirtuin) deacetylases is conserved in all domains of life. In the prokaryote Salmonella enterica, the metabolic enzyme acetyl-coenzyme A synthetase (Acs) is regulated by a Sir2-dependent protein acetylation/deacetylation system (SDPADS). The recent identification of the acetyltransferase enzyme responsible for the acetylation of Acs defined the SDPADS in prokaryotes. This report identifies one residue in Acs, Leu-641, which is critical for the acetylation of Acs by the protein acetyltransferase enzyme. In vivo and in vitro evidence shows that mutations at Leu-641 prevent the acetylation of Acs by protein acetyltransferase, maintain the Acs enzyme in its active state, and bypass the need for sirtuin deacetylase activity during growth on acetate. PMID- 15899898 TI - Tissue-specific changes in H19 methylation and expression in mice with hyperhomocysteinemia. AB - Expression of the imprinted genes H19 and insulin-like growth factor 2 (Igf2), which lie in close proximity on mouse chromosome 7, is regulated by methylation of a differentially methylated domain (DMD) located 5' to H19. Biallelic expression of H19 has been observed in renal disease patients with hyperhomocysteinemia, a cardiovascular disease risk factor. The present study determined whether hyperhomocysteinemia produces decreased tissue methylation capacity, hypomethylation of the H19 DMD, and altered expression of H19 and Igf2 in adult mice. Mice heterozygous for disruption of the gene for cystathionine beta-synthase (Cbs+/-) and C57BL/6 (Cbs+/+) mice were fed a hyperhomocysteinemic or control diet, respectively, from weaning until 9-12 months of age. Higher plasma total homocysteine (p < 0.001) was found in hyperhomocysteinemic mice than in control mice (95 +/- 12 versus 5.0 +/- 0.3 micromol/liter). Hyperhomocysteinemia was accompanied by higher levels of S-adenosylhomocysteine (p < 0.05) and lower S-adenosylmethionine/S-adenosylhomocysteine ratios (p < 0.001) in liver and brain. The effect of hyperhomocysteinemia on H19 DMD methylation was tissue-specific. In liver, hyperhomocysteinemic mice had decreased H19 DMD methylation (p < 0.001). In brain, hyperhomocysteinemia was accompanied by increased H19 DMD methylation (p < 0.001) and a decrease in the ratio of H19/Igf2 transcripts (p < 0.05). In aorta, hyperhomocysteinemia produced an increase in H19 DMD methylation (p < 0.001) and a 2.5-fold increase in expression of H19 transcripts (p < 0.05). Levels of H19 transcripts in aorta correlated positively with plasma total homocysteine concentration (p < 0.05, r = 0.620). We conclude that hyperhomocysteinemia produces tissue-specific changes in H19 DMD methylation and increased vascular expression of H19 in adult mice. PMID- 15899899 TI - N-Glycans of Caenorhabditis elegans are specific to developmental stages. AB - We have examined the N-glycans present during the developmental stages of Caenorhabditis elegans using two approaches, 1) a combination of permethylation followed by MALDI-TOF mass spectrometry (MS) and 2) derivatization with 2 aminobenzamide followed by separation by high-performance liquid chromatography and analyses by MALDI-TOF MS, post source decay (PSD) MS, and MALDI-QoTOF MS/MS. The N-glycan profile of each developmental stage (Larva 1, Larva 2, Larva 3, Larva 4, and Dauer and adult) appears to be unique. The pattern of complex N glycans was stage-specific with the general trend of number and abundance of glycans being Dauer approximately = L1 > adult approximately = L4 > L3 approximately = L2. Dauer larvae contained complex N-glycans with higher molecular masses than those seen in other stages. MALDI-QoTOF MS/MS of Hex4HexNAc4 showed an N-acetyllac-tosamine substitution not previously observed in C. elegans. Phosphorylcholine (Pc)-substituted glycans were also found to be stage-specific. Higher molecular weight Pc-containing glycans, including fucose containing ones such as difucosyl Pc-glycan (Pc1dHex2Hex5HexNAc6) seen in Dauer larvae, have not been observed in any organism. Pc2Hex4HexNAc3, from Dauer larvae, when subjected to PSD MS analyses, showed Pc may substitute both core and terminally linked GlcNAc; no such structure has previously been reported in any organism. C. elegans-specific fucosyl and native methylated glycans were found in all developmental stages. Taken together, the above results demonstrate that in depth investigation of the role of the above N-glycans during C. elegans development should lead to a better understanding of their significance and the ways that they may govern interactions, both within the organism during development and between the mobile nematode and its pathogens. PMID- 15899900 TI - A novel phospholipase C, PLC(eta)2, is a neuron-specific isozyme. AB - Twelve phospholipase C (PLC) isozymes have been cloned so far, and they are divided into six classes, beta-, gamma-, delta-, epsilon-, zeta-, and eta-type, on the basis of structure and activation mechanisms. Here we report the identification of a novel PLC isozyme, PLC(eta)2. PLC(eta)2 is composed of conserved domains including pleckstrin homology, EF-hand, X and Y catalytic, and C2 domains and the isozyme-specific C-terminal region. PLC(eta)2 consists of 1164 amino acids with a molecular mass of 125 kDa. The PLC activity of PLC(eta)2 was more sensitive to calcium concentration than the PLC activity of the PLCdelta type enzyme, which is thought to be the most calcium-sensitive PLC. Immunofluorescence analysis showed that PLC(eta)2 was localized predominantly to the plasma membrane at resting state via the pleckstrin homology domain. This observation was supported by Western blot analysis of cytosol and membrane fractions. In addition, expression of PLC(eta)2 was detected after birth and showed a restricted distribution in the brain; it was particularly abundant in the hippocampus, cerebral cortex, and olfactory bulb. The pattern was similar to that of the neuronal marker microtubule-associated protein 2 by Western blot. Furthermore, in situ hybridization showed positive signals for PLC(eta)2 in pyramidal cells of the hippocampus. Finally, we found that PLC(eta)2 was expressed abundantly in neuron-containing primary culture but not in astrocyte enriched culture. These results indicate that PLC(eta)2 is a neuron-specific isozyme that may be important for the formation and/or maintenance of the neuronal network in the postnatal brain. PMID- 15899901 TI - Disruption of fusion results in mitochondrial heterogeneity and dysfunction. AB - Mitochondria undergo continual cycles of fusion and fission, and the balance of these opposing processes regulates mitochondrial morphology. Paradoxically, cells invest many resources to maintain tubular mitochondrial morphology, when reducing both fusion and fission simultaneously achieves the same end. This observation suggests a requirement for mitochondrial fusion, beyond maintenance of organelle morphology. Here, we show that cells with targeted null mutations in Mfn1 or Mfn2 retained low levels of mitochondrial fusion and escaped major cellular dysfunction. Analysis of these mutant cells showed that both homotypic and heterotypic interactions of Mfns are capable of fusion. In contrast, cells lacking both Mfn1 and Mfn2 completely lacked mitochondrial fusion and showed severe cellular defects, including poor cell growth, widespread heterogeneity of mitochondrial membrane potential, and decreased cellular respiration. Disruption of OPA1 by RNAi also blocked all mitochondrial fusion and resulted in similar cellular defects. These defects in Mfn-null or OPA1-RNAi mammalian cells were corrected upon restoration of mitochondrial fusion, unlike the irreversible defects found in fzodelta yeast. In contrast, fragmentation of mitochondria, without severe loss of fusion, did not result in such cellular defects. Our results showed that key cellular functions decline as mitochondrial fusion is progressively abrogated. PMID- 15899902 TI - The human mitochondrial transcription termination factor (mTERF) is fully active in vitro in the non-phosphorylated form. AB - The human mitochondrial transcription termination factor (mTERF) is a 39-kDa protein that terminates transcription at the 3'-end of the 16 S rRNA gene and thereby controls expression of the ribosomal transcription unit of mitochondrial DNA. The transcription termination activity of human mTERF has been notoriously difficult to study in vitro, and it has been suggested that the activity of the protein is regulated by posttranslational modifications or by protein polymerization. We here characterize the activity of recombinant human mTERF expressed in insect cells. We observed that mTERF efficiently promotes sequence specific termination in a completely recombinant and highly purified in vitro system for mitochondrial transcription. The termination activity has a distinct polarity, and we observed complete transcription termination when the mTERF binding site is oriented in a forward position relative the heavy strand promoter but only partial transcription termination when the binding site is in the reverse position. We analyzed the biochemical characteristics of the active mTERF protein and found that it is a stable monomer at physiological salt concentration. Structural analysis, including phosphostaining, two-dimensional electrophoresis, and electrospray mass spectrometry, detected no evidence of phosphorylation. We conclude that the monomeric human mTERF is fully active in its non-phosphorylated form and that the protein does not require additional cellular factors to terminate mitochondrial transcription in vitro. PMID- 15899903 TI - Recognition of phospholipids in Streptomyces phospholipase D. AB - To investigate the contribution of amino acid residues to the enzyme reaction of Streptomyces phospholipase D (PLD), we constructed a chimeric gene library between two highly homologous plds, which indicated different activity in transphosphatidylation, using RIBS (repeat-length independent and broad spectrum) in vivo DNA shuffling. By comparing the activities of chimeras, six candidate residues related to transphosphatidylation activity were shown. Based on the above result, we constructed several mutants to identify the key residues involved in the recognition of phospholipids. By kinetic analysis, we identified that Gly188 and Asp191 of PLD from Streptomyces septatus TH-2, which are not present in the highly conserved catalytic HXKXXXXD (HKD) motifs, are key amino acid residues related to the transphosphatidylation activity. To investigate the role of two residues in the recognition of phospholipids, the effects of these residues on binding to substrates were analyzed by surface plasmon spectroscopy. The result suggests that Gly188 and Asp191 are involved in the recognition of phospholipids in correlation with the N-terminal HKD motif. Furthermore, this study also provides experimental evidence that the N-terminal HKD motif contains the catalytic nucleophile, which attacks the phosphatidyl group of the substrate. PMID- 15899904 TI - Prostaglandin E2 Stimulates the beta-catenin/T cell factor-dependent transcription in colon cancer. AB - Cyclooxygenase and its derived prostaglandin E2 (PGE2) have been shown to stimulate the growth of cancer cells and promote tumor angiogenesis. Here, we show that PGE2 activated the beta-catenin/T cell factor-dependent transcription in colon cancer cells through the cAMP/protein kinase A pathway. The expression of cyclin D1 and vascular endothelial growth factor was induced by PGE2 in LS 174T cells. Moreover, PGE2 and mutated beta-catenin stimulated the transcription of cyclin D1 and vascular endothelial growth factor in a synergistic fashion. Mechanistically, PGE2 increased the phosphorylation of glycogen synthase kinase-3 and consequently accumulated beta-catenin. In addition, PGE2 induced the expression of T cell factor-4 transcription factor, which formed transcriptionally active complex with beta-catenin. In animal experiments, administration of 16,16-dimethyl PGE2 strongly increased the expression of cyclin D1 and vascular endothelial growth factor in APC(min/+) mouse polyps. Thus, our results provide a novel mechanism, suggesting that cyclooxygenase-2/PGE2 may exert pro-oncogenic actions through stimulating the beta-catenin/T cell factor mediated transcription, which plays critical roles in colorectal carcinogenesis. PMID- 15899905 TI - Communicable Disease and Health Protection Quarterly Review: October to December 2004. From the Health Protection Agency, Centre for Infections. PMID- 15899906 TI - The legacy of monazite processing in Brazil. AB - The exploitation of natural resources containing naturally occurring radionuclides may lead to enhanced levels of radioactive isotope and enhanced potential for exposure to naturally occurring radionuclides in products, by products, residues or wastes. Such resources include, for instance, monazite, the processing of which, in Brazil, generated a great amount of radioactive residues, being stored in buried concrete tanks, in temporary storage buildings and in sealed trenches. In addition, during the 1980s there were no radiological protection rules concerning the storage and transportation of these kinds of residues. Mineral radioactive residues were used as landfills and the residues of chemical processes contaminated floors and buildings. The decommissioning process and cleaning of old plants have generated tons of wastes that has been added to previously produced wastes. This paper reports and discusses the cycle of monazite in Brazil and its consequences in terms of site remediation and amount of wastes and residues generated and stored. PMID- 15899907 TI - Effect of calcium trisodium DTPA in rats with puncture wound contaminated by 90Y chloride. AB - The efficacy of diethylenetriaminepentaacetate calcium trisodium (CaNa(3)DTPA) in a dose of 34.7 micromol kg(-1) as a function of its route of administration was investigated in rats with a puncture wound contaminated by (90)Y-chloride at a concentration of 2.55 MBq kg(-1). Approximately 60% of (90)Y-chloride at a puncture wound was absorbed into the body of rats over 72 h post-puncture and radioactivity in femoral bone increased during the timed-release of (90)Y. Intravenous administration of CaNa(3)DTPA (systemic treatment) at 15 min post puncture reduced (90)Y at a puncture wound and in bone up to 75.6 and 84.3% of controls, respectively. Direct infiltration of CaNa(3)DTPA into a puncture wound site (local treatment) at 15 min post-puncture diminished radioactivity at the puncture wound and in bone up to 34.9 and 52.5% of controls, respectively. Thus, prompt local treatment may be effective for removing (90)Y from a puncture wound and minimising (90)Y-distribution to bone compared with systemic treatment. PMID- 15899908 TI - Annual effective dose due to natural radionuclides in building blocks in eight cities of Southwestern Nigeria. AB - The specific absorbed dose rates due to (226)Ra, (232)Th and (40)K in building blocks have been determined for a model dwelling dimension of 3.6 x 3.6 x 3.0 m(3), wall thickness 10 cm and density of 1.73 x 10(3) kg m(-3) in eight cities across Southwestern Nigeria. The Mustonen method of calculating gamma-ray annual exposure rate from the radioactivity concentrations (Bq kg(-1)) of (226)Ra, (232)Th and (40)K in the blocks have been adopted with modifications to suit the Nigerian situation. The specific absorbed dose rates obtained were as follows: 11.36, 14.94 and 0.92 microGy y(-1) per Bq kg(-1) for (226)Ra, (232)Th and (40)K, respectively. These dose rates were used to calculate the annual effective dose in the model dwelling. The annual effective dose obtained varied between 209.20 (Osogbo) and 400.36 microSv y(-1) (Abeokuta). The values are in the range of values obtained for the German Democratic Republic (309 microSv y(-1)) and Jordan (470 microSv y(-1)). PMID- 15899909 TI - Changing age-pattern of hospitalisation risk of chronic obstructive pulmonary disease in men and women in Canada. AB - OBJECTIVE: To examine the changing pattern of age distributions of hospitalisation for chronic obstructive pulmonary disease (COPD) among Canadian men and women. DESIGN: Retrospective cohort study. PARTICIPANTS: 257,604 COPD inpatients aged 55-90 years with 463,089 hospital admissions during a 3-year study period (1994/95, 1995/96 and 1996/97) in Canada. MAIN OUTCOME MEASURES: COPD listed as one of the first five underlying diagnoses (broad definition, 463,089 hospitalisations) or as first diagnosis (narrow definition, 142,770 hospitalisations). RESULTS: Overall, men were more likely to have hospitalisations for COPD and had a higher proportion of death at hospital than did women. The 3-year cumulative incidence was 42.2/1,000 for the broadly defined COPD hospitalisation and 14.0/1,000 for the narrowly defined COPD hospitalisation, and steadily increased with increasing age. The relative risk for women versus men gradually increased with decreasing age, and was significantly greater than unity in the 55-59 year group for narrowly defined COPD hospitalisation. CONCLUSIONS: In terms of impact on secondary care COPD is a disease of the elderly and is becoming more common in women, particularly in younger age groups. PMID- 15899910 TI - Perceived age as a predictor of old age mortality: a 13-year prospective study. AB - OBJECTIVES: To examine whether in older people perceived age is associated with risk of total mortality independent of chronological age. DESIGN: Prospective population-based study (Evergreen project) with mortality surveillance for 13 years after the baseline. SETTING: Face-to-face interview among community dwelling residents of the city of Jyvaskyla, Finland. SUBJECTS: 395 men and 770 women aged 65-84 years at baseline. MEASURES: Perceived physical age and perceived mental age were rated either as younger, the same or older in comparison with subject's chronological age. Death dates were received from the official register of the province of Central Finland. Confounders used were chronological age, education in years, number of long-term illnesses, self-rated health, depression score (Beck's 13-item depression scale), and cognitive status. RESULTS: Mortality rates per 1,000 person-years from the older to younger perceived physical age category were 99, 65 and 59 in men, and 81, 54 and 36 in women. In the perceived mental age categories, correspondingly, mortality rates were 139, 63 and 64 in men, and 82, 55 and 44 in women. The fully adjusted relative risk (RR) of death over 13 years with the perceived younger physical age category as referent was 1.42 (95% CI 1.00-2.02) in the older category and 1.28 (1.03-1.60) in the same age category (P=0.049). The crude RR of mortality for perceived mental age categories was 1.56 (1.09-2.23) in the older and 1.10 (0.92 1.31) in the same age as compared with the younger category (P=0.046). Adding cognitive status into the model diminished the predictive value of the model (P=0.545). CONCLUSIONS: Perceived age predicted worsening of health as described as mortality. Perceived age may indicate general well-being and faith in the future, potentially reflecting changes in health. PMID- 15899911 TI - Association between sensorimotor function and functional and reactive balance control in the elderly. AB - OBJECTIVES: Postural disturbances can arise from performing functional tasks and from external perturbations. Identification of sensorimotor factors associated with both types of balance control in the elderly can help us to understand better the balance problems facing older adults. DESIGN: Cross-sectional. SUBJECTS: Healthy young, stable older, and functionally unstable older adults with 16 participants in each group. METHODS: Clinical vibration sense and muscle strength of the lower extremity, and functional balance (FB) tests were conducted. The timing and amplitude of the reactive postural muscle responses of the leg postural muscles, recorded from standing subjects following support surface backward translation, were also examined. RESULTS: Young and older subjects differed significantly in the amplitude of their postural muscle responses, while the two older groups differed significantly in muscle strength and FB. When age was controlled, the strength of the ankle dorsi- and plantar flexors was the only significant predictor for FB. For reactive postural muscle responses, none of the sensorimotor factors was significant. CONCLUSION: Functional and reactive balance abilities differed in their associating factors. The difference in the patterns of association for functional and reactive balance implies the need for separate assessment for these two categories of balance control clinically. PMID- 15899912 TI - Decreased PARP and procaspase-2 protein levels are associated with cellular drug resistance in childhood acute lymphoblastic leukemia. AB - Drug resistance in childhood acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) is associated with impaired ability to induce apoptosis. To elucidate causes of apoptotic defects, we studied the protein expression of Apaf 1, procaspases-2, -3, -6, -7, -8, -10, and poly(adenosine diphosphate [ADP] ribose) polymerase (PARP) in cells from children with acute lymphoblastic leukemia (ALL; n = 43) and acute myeloid leukemia (AML; n = 10). PARP expression was present in all B-lineage samples, but absent in 4 of 15 T-lineage ALL samples and 3 of 10 AML cases, which was not caused by genomic deletions. PARP expression was a median 7-fold lower in T-lineage ALL (P < .001) and 10-fold lower in AML (P < .001) compared with B-lineage ALL. PARP expression was 4-fold lower in prednisolone, vincristine and L-asparaginase (PVA)-resistant compared with PVA sensitive ALL patients (P < .001). Procaspase-2 expression was 3-fold lower in T lineage ALL (P = .022) and AML (P = .014) compared with B-lineage ALL. In addition, procaspase-2 expression was 2-fold lower in PVA-resistant compared to PVA-sensitive ALL patients (P = .042). No relation between apoptotic protease activating factor 1 (Apaf-1), procaspases-3, -6, -7, -8, -10, and drug resistance was found. In conclusion, low baseline expression of PARP and procaspase-2 is related to cellular drug resistance in childhood acute lymphoblastic leukemia. PMID- 15899913 TI - Efficient migration of dendritic cells toward lymph node chemokines and induction of T(H)1 responses require maturation stimulus and apoptotic cell interaction. AB - Dendritic cells (DCs) have the unique ability to initiate primary immune responses, and they can be conditioned for vaccinal purposes to present antigens after the engulfment of apoptotic cells. To recruit the rare antigen-specific naive T cells, DCs require a maturation step and subsequent transport toward lymph node (LN). To date, prostaglandin E2 (PGE2) is the best-characterized compound inducing this LN-directed migration in vitro, but PGE2 may skew the immune responses in a T(H)2 direction. We demonstrate here that on incubation with apoptotic tumor cells and tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS), human monocyte-derived DCs become fully mature and acquire high migratory capacities toward LN-directing chemokines. The migration of TNF-alpha-treated DCs occurs only after cotreatment with apoptotic cells but not with necrotic cells. DC migration requires CD36 expression and incubation with apoptotic cells in the presence of heat-labile serum components. Moreover, on treatment with apoptotic cells and LPS, the migrating DCs are able to recruit naive T cells to generate T(H)1 immune responses. Our results show that the cotreatment of DCs with apoptotic tumor cells and inflammatory signals is promising for the design of an antitumoral DC-based vaccine. PMID- 15899914 TI - Mouse Polycomb M33 is required for splenic vascular and adrenal gland formation through regulating Ad4BP/SF1 expression. AB - Mice with disrupted mammalian PcG (Polycomb group) genes commonly show skeletal transformation of anterior-posterior identities. Disruption of the murine M33 gene, a PcG member, displayed posterior transformation of the vertebral columns and sternal ribs. In addition, failure of T-cell expansion and hypoplasia and sex reversal of the gonads, have been observed. In the present study, we identified defects in the splenic and adrenal formation of M33-knock-out (KO) mice on a C57BL/6 genetic background. The spleen in these animals was smaller than in the wild-type mice and was spotted red because of nonuniform distribution of blood cells. Histologic examination revealed disorganization of the vascular endothelium and its surrounding structures, and immunohistochemistry demonstrated disturbances in vascular formation and colonization of immature hematopoietic cells. These splenic phenotypes observed in the M33-KO mice were quite similar to those seen in Ad4BP/SF1 (Nr5a1) knock-outs. Moreover, the adrenal glands of M33 KO and Ad4BP/SF1 heterozygous KO mice were smaller than those of the wild-type mice. Western blot, immunohistochemistry, and reverse transcriptase-polymerase chain reaction (RT-PCR) analyses of the M33 knock-outs all indicated significantly low expression of adrenal 4 binding protein/steroidogenic factor-1 (Ad4BP/SF-1), indicating that M33 is an essential upstream regulator of Ad4BP/SF1. In agreement with these observations, chromatin immunoprecipitation assays with adrenocortical Y-1 cells revealed direct binding of the M33 containing PcG to the Ad4BP/SF1 gene locus. PMID- 15899915 TI - Suppressor of cytokine signaling-1 in T cells and macrophages is critical for preventing lethal inflammation. AB - The balance between pro- and anti-inflammatory cytokines modulates inflammation. Intracellular inhibitors of signaling, in turn, contribute to the negative regulation of cytokines. One of these inhibitors is suppressor of cytokine signaling-1 (SOCS-1). Socs1(-/-) mice die by 3 weeks of age with inflammation and fatty necrosis of the liver. Here, cre/loxP deletion of Socs1 was used to investigate the contribution of specific cells/tissues to inflammatory disease. Mice with SOCS-1 deficiency in myeloid and lymphoid cells, but not lymphoid alone, became ill at 50 to 250 days of age. These mice developed splenomegaly and T-cell/macrophage infiltration of many organs, including liver, lung, pancreas, and muscle. There were also abnormally high levels of the proinflammatory cytokines interferon gamma (IFN-gamma), tumor necrosis factor (TNF), and interleukin-12 (IL-12), and activated T cells circulating in these mice. Socs1(null) T cells were found to be hypersensitive to multiple cytokines, including IL-1, IL-2, and IL-12, resulting in IFN-gamma production without requiring T-cell receptor (TCR) ligation. Additionally, Socs1(null) macrophages produced excessive amounts of IL-12 and TNF in response to other cytokines, including IFN-gamma. A dysregulated cytokine network between T cells and macrophages is thus associated with this inflammatory disease. These findings indicate that SOCS-1 is critical in both T cells and macrophages for preventing uncontrolled inflammation. PMID- 15899916 TI - Rapid immunosuppressive effects of glucocorticoids mediated through Lck and Fyn. AB - Glucocorticoids (GCs) are effective immunosuppressive agents and mediate well defined transcriptional effects via GC receptors. There is increasing evidence that GCs also initiate rapid nongenomic signaling events. Using activated human CD4(+) lymphocytes and a peptide array containing 1176 different kinase consensus substrates, we generated a comprehensive profile of GC-induced rapid effects on signal transduction. The results show marked early differences in phosphorylation between GC-pretreated cells and control cells, including impaired phosphorylation of p56lck/p59fyn (Lck/Fyn) consensus substrates. Immunoprecipitation and in vitro kinase assays reveal rapid GC-induced down-modulation of Lck and Fyn kinases using SAM68 (Src [pp60c-src]-associated in mitosis 68 kDa) as a substrate. Additionally, immunoprecipitation experiments revealed reduced Lck-CD4 and Fyn CD3 associations, suggesting GC inhibited recruitment of these kinases to the T cell receptor complex. Western blot analysis revealed reduced phosphorylation of a series of downstream signaling intermediates following GC treatment, including protein kinase B (PKB), protein kinase C (PKC), and mitogen-activated protein kinases (MAPKs). Experiments with GC receptor-negative Jurkat cells and a pharmacologic GC receptor ligand (RU486) indicated that rapid inhibition of Lck and Fyn kinases is GC receptor dependent. Parallel experiments conducted following the application of GCs in healthy individuals confirmed suppression of Lck/Fyn in T cells within 1 hour in vivo. These results identify the inhibition of Lck and Fyn kinases as rapid targets of GCs, mediated via a GC receptor dependent pathway. PMID- 15899917 TI - Identification of NKG2A and NKp80 as specific natural killer cell markers in rhesus and pigtailed monkeys. AB - Investigations of natural killer (NK) cells in simian models of disease have been hampered by a lack of appropriate phenotypic markers and by an inadequate understanding of the regulation of NK cell activities. In the present study, a panel of monoclonal antibodies (mAbs) specific for various human NK receptors was screened for cross-reactivity with NK cells from rhesus macaques and pigtailed macaques. Flow cytometric analyses using anti-human NKG2A and anti-human NKp80 mAbs individually, and particularly in combination with anti-CD16 mAb, allowed for the identification of the entire NK cell population in both species. NK cells in monkeys were generally identified by negative selection of peripheral blood mononuclear cells (PBMCs) for the absence of T-cell, B-cell, and monocyte markers. mAb-mediated ligation of NKp80 induced NK cell cytotoxicity, while in the case of NKG2A it displayed a clear capability to inhibit the lysis of target cells by NK cells from macaques, as well as from humans. This new phenotypic and functional characterization of NKG2A and NKp80 in rhesus and pigtailed macaque NK cells provides a new approach in the analysis of their innate immune system. PMID- 15899918 TI - Tolerance induction by bone marrow transplantation in a multiple sclerosis model. AB - Experimental autoimmune encephalomyelitis (EAE) in rats is a highly valuable model of multiple sclerosis (MS) because it mimics major hallmarks of the human disease. EAE induced with myelin-oligodendrocyte-glycoprotein (MOG) in DA rats is relapsing/remitting, and lesions in the central nervous system show inflammation, demyelination, and axonal and neuronal loss. Recently, bone marrow transplantation (BMT) was introduced as a novel strategy to treat MS, but its efficiency and the underlying mechanism are debatable. In MOG-induced EAE we found that BMT at the peak of EAE but not in the chronic phase leads to disease attenuation. In both settings, rats receiving bone marrow (BM) transplants were protected from subsequently induced relapses. These findings could be confirmed by histopathology in which rats receiving BM transplants did not have lesions compared with controls not receiving transplants. Importantly, the protective effect was achieved by allogeneic, syngeneic, and BM grafts from diseased rats. BMT resulted in increased numbers of CD4(+)CD25(bright) regulatory T cells, increased Foxp3 expression, a shift in T-cell epitope recognition, and a strong reduction of autoantibodies even after rechallenge with MOG. Thus, our results indicate potential mechanisms of how BMT may contribute to the improvement of MS and provide a rationale for its application in patients suffering from various autoimmune diseases. PMID- 15899919 TI - Balanced expression of CXCR5 and CCR7 on follicular T helper cells determines their transient positioning to lymph node follicles and is essential for efficient B-cell help. AB - The production of high-affinity antibodies to T-dependent antigens requires the interaction of B cells and T helper cells expressing receptors specific for the same antigen. Although several mechanisms have been elucidated that regulate B cell trafficking within lymphoid organs, less is known about molecular cues that guide the small subpopulation of CD4+ follicular T helper cells to B-cell follicles. Using adoptive transfer of transgenic T cells in mice, we demonstrate that antigen-induced activation leads to a finely tuned positioning of T cells either to the T-cell area or the B-cell follicle. We show that expression of CXCR5 is indispensable for T cells to enter B-cell follicles, whereas expression of CCR7 provides a counteracting signal to retain activated T cells in the T-cell area. Although only few T cells transiently migrate from the T-cell area to the B cell follicle of peripheral lymph nodes following antigenic challenge, this step is essential to provide the help B cells require to produce antibodies efficiently. Thus, we demonstrate that the balanced expression of CCR7 and CXCR5 determines the positioning and proper function of follicular T helper cells. PMID- 15899920 TI - Bcl-x(L) prevents apoptosis of late-stage erythroblasts but does not mediate the antiapoptotic effect of erythropoietin. AB - The long form of B-cell lymphoma-x (Bcl-x(L)), an outer mitochondrial membrane protein, has been proposed to mediate the antiapoptotic action of erythropoietin on erythroid progenitor cells and to be necessary for heme synthesis in erythroblasts. Mice with conditional knockout of Bcl-x(L) (conditional bcl-x(-/-) mice) develop severe anemia that has been attributed to hemolysis and is accompanied by splenomegaly. We characterized further the anemia of conditional bcl-x(-/-) mice and investigated the role of Bcl-x(L) in the action of erythropoietin and in heme synthesis. We analyzed peripheral blood cells and cultured splenic erythroblasts of conditional bcl-x(-/-) mice and littermates that were rendered anemic by bleeding. Although they had massive splenic erythroblastosis, conditional bcl-x(-/-) mice had decreased circulating reticulocytes compared to littermates even prior to bleeding the littermates. Compared to erythroblasts of bled littermates, bcl-x(-/-) erythroblasts cultured with erythropoietin underwent apoptosis during the later, hemoglobin-synthesizing stages of differentiation. The bcl-x(-/-) erythroblasts synthesized heme, but at reduced rates compared to bled littermate erythroblasts. When cultured without erythropoietin, bcl-x(-/-) erythroblasts underwent apoptosis at early stages of differentiation, prior to hemoglobin synthesis. Bcl-x(L) is not required for heme synthesis and does not mediate the antiapoptotic effects of erythropoietin, but it prevents ineffective erythropoiesis due to apoptosis in late-stage, hemoglobin synthesizing erythroblasts. PMID- 15899921 TI - Randomised trial of parental support for families with very preterm children: outcome at 5 years. AB - AIMS: To test the effectiveness of a home based developmental education intervention in improving outcome at 5 years for very preterm infants. METHODS: The Avon Premature Infant Project (APIP) is a randomised controlled trial in which the parents of 284 babies born <33 weeks gestational age received a developmental education programme, a social support intervention, or standard care. A term reference population was also recruited. This study reports outcomes at 5 years (mean age 58 months 15 days) for 187 (66%) of these children without disability. Outcomes were assessed using the British Ability Scales II for cognitive development, the Movement ABC for motor impairment, and the Child Behavior Checklist for behavioural problems. RESULTS: Preterm infants showed poorer cognitive performance than their term peers. Mean (SD) general conceptual ability (GCA) scores were: Portage 99.2 (15.7); parent adviser 100.3 (14.8); preterm control 101.1 (15.0); term reference 107.2 (13.4). There were no significant differences between preterm groups in GCA scores indicating no effect of either intervention. Similarly, there was no significant effect of intervention on behavioural or motor outcomes. Further analyses, in which outcome data were adjusted for social factors, did not reveal any differences between the three preterm groups or by subgroups classified by a range of perinatal variables. CONCLUSION: The small advantage shown at 2 years of age is no longer detectable at 5 years. These results question the effectiveness of early intervention in enhancing cognitive, behavioural, and motor function at 5 years. PMID- 15899922 TI - Children with unexplained chronic pain: substantial impairment in everyday life. AB - AIMS: To describe and quantify impairment in an outpatient population of children with chronic pain of unknown origin (UCP). METHODS: A total of 149 children who presented with pain of at least three months' duration and without a satisfactory explanation at presentation were studied. Number of somatic complaints (Children's Somatisation Inventory, CSI), pain intensity (VAS, 0-10 cm), functional disability (Child Health Questionnaire (CHQ-CF) and clinical history), and general health perceptions (CHQ) were determined. RESULTS: Mean age of the children was 11.8 years; 73% were girls. Overall, 72% suffered impairment in sports activities, 51% reported absence from school, 40% experienced limitations in social functioning, and 34% had problems with sleeping. Mean number of somatic symptoms differed significantly between boys (8.4) and girls (10.7). The CHQ-CF scores for physical functioning, role/social functioning, and general health perceptions were 76.4, 70.7, and 57.5, respectively, indicating substantial impairment on all domains. The mean pain intensity was 4.7 for current and 7.1 for worst pain. Children solely evaluated by a general practitioner prior to referral reported less, though still substantial, impairment. Low general health perceptions, impaired role/social functioning, high pain intensity, and having headache or musculoskeletal pain were independent predictors of having significant impairment. CONCLUSIONS: Referred children with UCP show substantial impairment on multiple domains in daily life. PMID- 15899923 TI - Induction and activation of the aryl hydrocarbon receptor by IL-4 in B cells. AB - It is widely known that IL-4 and IL-13 act on various kinds of cells, including B cells, resulting in enhancement of proliferation, class switching to IgE and expression of several surface proteins. These functions are important for the recognition of the various antigens in B cells and are known to be involved in the pathogenesis of allergic diseases. However, it has not been known whether IL 4/IL-13 is involved in the metabolism of various kinds of xenobiotics including 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD), and it remains undetermined whether TCDD, an environmental pollutant, influences IgE production in B cells, exaggerating allergic reactions. We identified IL-4- or IL-13-inducible genes in a human Burkitt lymphoma cell line, DND-39, using microarray technology, in which the AHR gene was included. The AHR gene product, the aryl hydrocarbon receptor (AhR), was induced by IL-4 in both mouse and human B cells in a STAT6-dependent manner. IL-4 alone had the ability to translocate the induced AhR to the nuclei. TCDD, a ligand for AhR, rapidly degraded the induced AhR by the proteasomal pathway, although IL-4-activated AhR sustained its expression. AhR activated by IL-4 caused expression of a xenobiotic-metabolizing gene, CYP1A1, and TCDD synergistically acted on the induction of this gene by IL-4. However, the induction of AhR had no effect on IgE synthesis or CD23 expression. These results indicate that the metabolism of xenobiotics would be a novel biological function of IL-4 and IL-13 in B cells, whereas TCDD is not involved in IgE synthesis in B cells. PMID- 15899924 TI - Prolonged skin allograft survival by IL-10 gene-introduced CD4 T cell administration. AB - Both CD4 and CD8 T cells play crucial roles in immune responses in transplantation. Immunosuppressive drugs, such as FK506 and cyclosporin A, block the priming of alloreactive CD4 T(h) cells and the subsequent induction of allospecific CD8 cytotoxic effector T cells and inhibit allograft rejection. However, the desire to minimize chronic complications that may arise from the use of immunosuppressive agents drives the search for additional strategies for immunosuppression of allograft rejection. In this study, CD4 or CD8 T cells into which the IL-10 gene is introduced using an adenovirus vector containing human IL 10 (hIL-10) cDNA (Ad-hIL-10) and into mouse T cells transgenic for the Coxsackie virus and adenovirus receptor form a model system to study the effect of administration of IL-10-secreting T cells on the survival of the allogenic skin grafts. Ad-hIL-10-infected CD4 and CD8 T cells secreted a large amount of hIL-10 for 3-4 days in culture in vitro. Ad-hIL-10-infected CD4 T cells administered in vivo could be detected in the spleen for 7 days post-transfer. Significantly prolonged survival of grafts was observed in animals that received either Ad-hIL 10-infected activated CD4 T cells or T(h)2-skewed CD4 T cells as compared with controls. Furthermore, substantial enhancement of the effect was observed in B6.C H2(bm1)/ByJ transplants. Thus, a direct manipulation of T cells through the introduction of the immunosuppressive cytokine gene IL-10 may be a novel strategy for the control of allograft rejection. PMID- 15899925 TI - Immunosynapse formation coincides with rapid activation of NK cells by syngeneic T cells and correlates with clustering of MHC class I. AB - T cells cultured for 3 h with antigen-presenting cells (APCs) stimulated syngeneic IL-2-activated NK cells as measured via a standard chromium-release assay. Discrete caps containing both TCR and MHC-I had formed on the surface of these activated T cells. When conjugates were formed between NK cells and these activated T cells, >80% of the contact sites were in the MHC-I(dim) region outside the TCR-MHC-I cap. Stimulation with phorbol myristate acetate plus Ionomycin, which bypasses the need for cell surface events during activation, did not induce either cap formation or NK cell activation. Further, the addition of the protein transport inhibitor Brefeldin A did not block activation of NK cells. MHC-I is the major inhibitory ligand recognized by NK cells. One possible mechanism for the activation of NK cells by TCR-MHC-I-capped T cells is that aggregation of MHC-I into one region leaves the remaining T cell surface denuded of ligands for NK-inhibitory receptors. As a test of this hypothesis, we aggregated MHC-I on T cells with plate-bound anti-MHC-I mAb. This treatment conferred upon the T cells the capacity to activate NK cells, suggesting that MHC I clustering could contribute to the observed phenomenon. PMID- 15899926 TI - Co-infection with Trypanosoma brucei brucei prevents experimental autoimmune encephalomyelitis in DBA/1 mice through induction of suppressor APCs. AB - The immune system has co-evolved with the infectious agents that challenge it, and in response pathogens have developed different mechanisms to subvert host immunity. A wealth of evidence suggests that infections are important components in the development of a functional immune system, and understanding the modulation of the host immune system by pathogens may offer new therapeutic strategies in a non-infectious setting. We investigated how infection with the protozoan parasite Trypanosoma brucei brucei (Tbb) modulates the autoimmune response to recombinant myelin oligodendrocyte glycoprotein (rMOG) in DBA/1 mice. Mice harbouring a Tbb infection did not develop experimental autoimmune encephalomyelitis (EAE) induced by immunization with rMOG in CFA, an animal model for the human autoimmune disease multiple sclerosis. Additionally, mice infected with the parasite at the time of immunization or 1 week later developed less severe EAE than uninfected controls. Protected mice displayed a markedly diminished rMOG-specific proliferation and IFNgamma production in lymph node cells and had correspondingly low titres of serum anti-rMOG IgG. Antigen presenting cells (APCs) from spleens of Tbb-infected mice presented rMOG less efficiently to rMOG-specific T cells in vitro than did splenic APCs from uninfected mice and could also inhibit antigen-specific proliferation in control in vitro cultures. This suppressive effect is at least in part due to increased release of IL-10. Transfer of splenic APCs from Tbb-infected mice into mice immunized with rMOG-CFA 7 days previously abrogated disease significantly. These findings indicate that infections can prevent autoimmunity and that APCs might be used as immunomodulants. PMID- 15899927 TI - Contrast enhanced-cardiovascular magnetic resonance imaging in patients with pulmonary hypertension. AB - AIMS: To determine the presence and extent of delayed contrast enhancement (DCE) in patients with pulmonary hypertension (PHT) using contrast enhanced cardiovascular magnetic resonance imaging (ce-CMR). METHODS AND RESULTS: Twenty five patients with PHT underwent ce-CMR and right heart catheterization. Right ventricular (RV) and left ventricular (LV) volumes, ejection fraction, mass, and DCE mass were determined with ce-CMR. Mean pulmonary artery pressure (mean PAP) averaged 43 (12) mmHg and cardiac output 4.3 (1.2) L/min. DCE was demonstrated in 23 out of 25 patients. DCE was confined to the RV insertion points (RVIPs) in seven patients and extended into the interventricular septum (IVS) in the remaining 16 patients. In these 16 patients, septal contrast enhancement was associated with IVS bowing. The extent of contrast enhancement correlated positively with RV end-diastolic volume/body surface area, RV mass, mean PAP, and pulmonary vascular resistance and correlated inversely with RV ejection fraction. CONCLUSION: DCE was present within the RVIPs and IVS of most patients with PHT studied. Extent of DCE correlated with RV function and pulmonary haemodynamics. DCE was associated with IVS bowing and may provide a novel marker for occult septal abnormalities directly relating to the haemodynamic stress experienced by these patients. PMID- 15899928 TI - Quinidine plus verapamil vs. quinidine alone to prevent recurrences of atrial fibrillation. PMID- 15899929 TI - Erythromycin as a prokinetic agent in preterm neonates: a systematic review. AB - BACKGROUND: It often takes several days or even weeks to establish full enteral feeds (FEFs) in preterm, especially extremely low birthweight neonates because of feed intolerance related to gastrointestinal hypomotility. Clinical trials of erythromycin as a prokinetic agent in preterm neonates have reported conflicting results. AIM: To systematically review the efficacy and safety of erythromycin as a prokinetic agent in preterm neonates. METHODS: Only randomised controlled trials in preterm neonates (gestation < or = 37 weeks) were considered eligible for inclusion. The primary outcome was the time to reach FEFs of 150 ml/kg/day. The secondary outcomes included the incidence of erythromycin related adverse effects such as diarrhoea, cardiac arrhythmias, and hypertrophic pyloric stenosis. No restrictions were applied on the dose (low: 3-12 mg/kg/day; antimicrobial: > or = 12 mg/kg/6-8 hours) and route (oral or intravenous) and mode (prophylactic or rescue) of administration. The standard methodology for systematic reviews was followed. A subgroup analysis was pre-planned based on the dose and mode of drug administration. RESULTS: Seven trials (three prophylaxis, four rescue) with various doses, routes and modes of administration, and durations of erythromycin treatment and different results were found to be eligible for inclusion in the analysis. Meta-analysis could not be performed, as specific data were either inadequate or not available. CONCLUSION: The conflicting trial results may be explained by differences in dose and route and mode of administration of erythromycin and in gastrointestinal motor responses in the presence of different feeding conditions-for example, fasting v fed state, intermittent v continuous feeds. Gestational and postnatal ages during erythromycin treatment are also important. PMID- 15899930 TI - Improved neurosensory outcome at 8 years of age of extremely low birthweight children born in Victoria over three distinct eras. AB - AIM: To determine neurosensory outcome at 8 years of age of extremely low birthweight (ELBW) children born in the 1990s, how it varies with birth weight, and how it compares with ELBW children born in the 1980s and 1970s. METHODS: Subjects were born in the state of Victoria during 1991-92 and comprised 224 of 241 consecutive survivors with a birth weight of 500-999 g, and 223 of 262 normal birthweight (NBW) controls who had been randomly selected at birth. The comparison cohorts from earlier eras comprised 87 of 89 consecutive ELBW survivors born in 1979-80, 206 of 212 consecutive ELBW survivors born in 1985-87, and 51 of 60 randomly selected NBW survivors born in 1981-82. Survivors were assessed for neurological impairments (cerebral palsy, blindness, deafness, intellectual impairment) and disabilities at 8 years of age by paediatricians and psychologists blinded to perinatal details. RESULTS: For the 1991-92 cohorts, the rate of neurosensory disability was substantially higher in ELBW children compared with NBW controls (p<0.0001). Within the ELBW group, neurosensory disability was more prevalent in children with a birth weight of <750 g compared with a birth weight of 750-999 g (p = 0.024). Disability rates were lower in the 1991-92 ELBW cohort compared with the ELBW cohort born in 1979-80 (p = 0.046). CONCLUSIONS: Neurosensory disability rates at school age were more common in ELBW children born in the 1990s compared with NBW controls, and were significantly more common in the 500-749 g birthweight subgroup, but have improved compared with ELBW children born in earlier eras. PMID- 15899931 TI - Internal contamination of gloves: routes and consequences. AB - The effect of internal glove contamination was investigated using N-methyl pyrrolidone (NMP) as a biological marker to assess systemic absorption when wearing internally contaminated gloves, and when not wearing gloves but subjected to the same challenge contaminant. The routes by which the insides of gloves become contaminated were also investigated. The area of dermal contamination was quantified using a fluorescent tracer dye and a surface monitoring fluorimeter. The main routes of internal glove contamination were found to be self contamination, cuff entry and failed gloves. Wearing internally contaminated gloves led to higher systemic absorption than was gained from the equivalent skin contamination when not wearing gloves. Repeat wetting of fingers with aqueous NMP, when gloves were not worn, gave higher systemic absorption than the equivalent continuous exposure, probably due to the low volatility of NMP leading to increased concentration and longer residence time on the skin. PMID- 15899932 TI - Detecting DNA repair capacity of peripheral lymphocytes from cancer patients with UVC challenge test and bleomycin challenge test. AB - The objective of this study was to evaluate DNA repair capacity of cancer patients with the bleomycin (BLM) challenge test and the UVC challenge test. The human peripheral lymphocytes were collected from 33 patients with different kinds of cancers and 33 controls in the same hospital. The lymphocytes of each subject were divided into two groups: (1) In the BLM challenge test, the lymphocytes were treated with BLM (20 microgml(-1)) for 30 min, and repaired for 15 min. The DNA damage before and after BLM exposure was detected with comet assay to assess DNA repair capacity. (2) In the UVC challenge test, the lymphocytes were exposed to UVC (254 nm) at the dose of 1.5 Jm(-2). DNA damage of lymphocytes was measured before UVC exposure and at 90 and 240 min after UVC exposure using comet assay, then DNA repair percentage (DRP) was calculated. The results of this study indicate that the average DRPs of cancer patients were 75.63 +/- 3.11 and 68.98 +/- 4.19% calculated with tail length (TL) and tail moment (TM), respectively, in the BLM challenge test, which were significantly lower than those (91.11 +/- 1.09 and 88.19 +/- 1.71%) of controls (P < 0.01). Also, the mean DRPs of cancer patients were 49.19 +/- 3.47 and 58.27 +/- 3.64% calculated with TL and TM, respectively, in the UVC test, which were significantly lower than those (77.52 +/- 2.06 and 83.12 +/- 2.36%) of controls (P < 0.01). The correlation between the DRPs (%) drawn with TL and TM in the BLM test or between the DRPs (%) drawn with mean TL and mean TM in the UVC challenge test were significant (P < 0.05). The DNA repair capacity measured with the BLM and UVC challenge tests in 33 cancer patients was significantly lower than that in controls. PMID- 15899933 TI - The in vivo comet assay: use and status in genotoxicity testing. AB - The in vivo comet assay (single cell gel electrophoresis assay) in its alkaline version (pH >13) is being increasingly used in genotoxicity testing of substances such as industrial chemicals, biocides, agrochemicals, food additives and pharmaceuticals. Recommendations for an appropriate performance of the test using OECD guidelines for other in vivo genotoxicity tests have been published. In this review, we critically discuss the biological significance of comet assay effects in general and the status of the test in current strategies for genotoxicity testing. Examples for practical applications of the in vivo comet assay and potential consequences of positive and negative test results are given. The significance of comet assay results for hazard identification and risk assessment is discussed. In accordance with international guidelines for genotoxicity testing the in vivo comet assay is recommended for follow-up testing of positive in vitro findings. It is particularly useful as a tool for the evaluation of local genotoxicity, especially for organs/cell types which cannot easily be evaluated with other standard tests. A positive result in an appropriately performed in vivo comet assay indicates genotoxicity of the test compound in the tissue tested and gains particular significance when a mutagenic potential of the test compound has already been demonstrated in vitro. Such findings will have practical consequences in the risk assessment processes and further development of substances. PMID- 15899934 TI - Validation of Drosophila melanogaster as an in vivo model for genotoxicity assessment using modified alkaline Comet assay. AB - The single cell gel electrophoresis or Comet assay is one of the most popular techniques for genotoxicity assessment. The present study was undertaken to validate our previously modified version of the Comet assay for genotoxicity assessment in Drosophila melanogaster (Oregon R(+)) with four well-known mutagenic and carcinogenic alkylating agents, i.e. ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), N-ethyl-N-nitrosourea (ENU) and cyclophosphamide (CP). Third instar larvae (74 +/- 2 h) of D.melanogaster were fed different concentrations of EMS, MMS, ENU and CP (0.05, 0.5 and 1.0 mM) mixed standard Drosophila food for 24 h. At 98 +/- 2 h, the anterior midgut from control and treated larvae were dissected out, single-cell suspensions were prepared and Comet assay was performed. Our results show a dose-dependent increase in DNA damage with all the four alkylating agents, in comparison to control. The lower concentration (0.05 mM) of the test chemicals, except MMS, did not induce any DNA damage in the gut cells of the exposed larvae. When comparison of Comet parameters was made among the chemicals, MMS was found to be the most potent genotoxicant and ENU the least. The present study validated our previous observation and shows that D.melanogaster is a sensitive and suitable model for the in vivo assessment of genotoxicity using our modified alkaline Comet assay. PMID- 15899935 TI - Associations of serum fetuin-A with malnutrition, inflammation, atherosclerosis and valvular calcification syndrome and outcome in peritoneal dialysis patients. AB - BACKGROUND: Fetuin-A (alpha2-Heremans Schmid glycoprotein) has recently been identified as a circulating inhibitor of calcification and is regulated as a negative acute phase protein. However, its relationships with cardiac valvular calcification and atherosclerosis and outcome have not been evaluated in peritoneal dialysis (PD) patients. METHOD: We performed a prospective follow-up study in 238 PD patients with echocardiography done at baseline to detect cardiac valvular calcification and biochemical analysis performed for serum fetuin-A, albumin and C-reactive protein (CRP). RESULTS: Baseline serum fetuin-A concentration was (mean+/-SD) 0.309+/-0.068 g/l (normal range 0.4-0.95). Across the three tertiles of increasing serum fetuin-A, a significant trend effect was observed for age (P = 0.023), diabetes (P = 0.008), background atherosclerotic vascular disease (P = 0.010), cardiac valvular calcification (P = 0.002), serum albumin (P<0.001), subjective global assessment (P = 0.005) and CRP (P<0.001). Adjusting for CRP and calcium x phosphorus product, every 0.01 g/l increase in serum fetuin-A remained independently associated with a 6% decrease in the risk of valvular calcification (95% confidence intervals, 0.90-0.99; P = 0.028). Furthermore, serum fetuin-A showed a significant decrease across the four groups of patients with increasing components of the malnutrition, inflammation, atherosclerosis/calcification (MIAC) syndrome (P<0.001) and was the lowest among patients with all components of the MIAC syndrome (0.263+/-0.055 g/l) and highest among those who do not have the MIAC syndrome at all (0.338+/-0.063 g/l). Lower serum fetuin-A was associated with greater all-cause mortality (P = 0.0011) and fatal and non-fatal cardiovascular events (P = 0.0017), but its significance was lost when atherosclerotic vascular disease, valvular calcification, inflammation and malnutrition were included in the model. CONCLUSIONS: Serum fetuin-A showed important associations with valvular calcification, atherosclerosis, malnutrition and inflammation, and was linked to mortality and cardiovascular events in PD patients via its close relationships with the MIAC syndrome. PMID- 15899936 TI - Henoch-Schonlein purpura in a patient with diabetic nephropathy and vascular complications. PMID- 15899937 TI - High clotting factor activities and enhanced platelet-related haemostasis in a patient with recurrent episodes of thrombotic-thrombocytopenic purpura: additional risk factors for onset and recurrence? PMID- 15899938 TI - Peritonitis: limiting the damage. PMID- 15899939 TI - The association between BP and mortality in patients on chronic peritoneal dialysis. AB - BACKGROUND: The relationship between blood pressure (BP) and mortality in haemodialysis patients is unconventional. It is not clear if this is the consequence of uraemia or related to the dialysis type. The goal of this project was to identify the relationship between BP and mortality in patients on chronic peritoneal dialysis (PD). METHODS: Patients on PD (n = 1053) from the USRDS prospective DMMS Wave 2 study were analysed. Primary outcomes were all-cause and cardiovascular mortality and duration of hospitalization. RESULTS: Low systolic BP categories, <100 mmHg [hazard ratio (HR), 2.71, P<0.001; and HR 3.83, P<0.001, respectively] and 101-110 mmHg (HR 1.85, P<0.05; and HR 2.92, P<0.005, respectively), but not high systolic BP, increased the risk of all-cause and cardiovascular mortality, with systolic BP 111-120 mmHg as the reference. Pulse BP, but not diastolic BP, followed a similar trend. In subgroup analysis, this association was demonstrated only in patients with a history of heart failure, in patients with diabetes and in those treated with antihypertensive medications. CONCLUSION: Systolic BP <111 mmHg in PD patients is associated with higher mortality risk, while systolic BP >120 mmHg is associated with fewer hospital days. Aggressive treatment of hypertension in the PD population should be cautioned. PMID- 15899940 TI - Interleukin-1beta and tumor necrosis factor-alpha mediation of endotoxin action on growth hormone. AB - In humans and sheep, endotoxin (LPS) administration results in increased growth hormone (GH) concentrations. To determine the role of cytokines in the effect of LPS on GH, sheep were challenged with IL-1beta or TNF-alpha. GH data were compared with results with LH, where the major effects of LPS are known to act via the hypothalamus. Intracerebroventricular (icv) administration of IL-1beta or TNF-alpha did not alter plasma concentrations of GH. Endotoxin was then administered intravenously (iv) in combination with icv injection of IL-1 receptor antagonist (IL-1RA), TNF antagonist (sTNF-R1), or saline. Administration of LPS increased GH (P < 0.0001), although coadministration of IL-1ra or sTNF-R1 icv did not alter GH response to LPS. In contrast, plasma concentrations of LH were profoundly inhibited by icv administration of either cytokine (P < 0.03), but the LH response to LPS was not altered by cytokine antagonists. Intravenous administration of either IL-1beta or TNF-alpha increased plasma concentrations of GH (P < 0.0001). Administration of IL-1RA and sTNF-R1 iv prevented LPS-induced increases in GH. Although LH was suppressed by high iv doses of IL-1beta (P = 0.0063), the antagonists did not alter the LH response to LPS. To determine whether LPS might directly activate GH release, confocal microscopy revealed colocalization of CD14, the LPS receptor, with GH and, to a lesser extent, LH and some prolactin (PRL)-containing cells, but not ACTH or TSH. These data are consistent with the effects of LPS on GH secretion originating through peripheral cytokine presentation to the pituitary, as well as a potential to act directly on selective populations of pituitary cells via CD14. PMID- 15899941 TI - Genetic background determines the extent of islet amyloid formation in human islet amyloid polypeptide transgenic mice. AB - Genetic background is important in determining susceptibility to metabolic abnormalities such as insulin resistance and beta-cell dysfunction. Islet amyloid is associated with reduced beta-cell mass and function and develops in the majority of our C57BL/6J x DBA/2J (F(1)) male human islet amyloid polypeptide (hIAPP) transgenic mice after 1 yr of increased fat feeding. To determine the relative contribution of each parental strain, C57BL/6J (BL6) and DBA/2J (DBA2), to islet amyloid formation, we studied male hIAPP mice on each background strain (BL6, n = 13; and DBA2 n = 11) and C57BL/6J x DBA/2J F(1) mice (n = 17) on a 9% (wt/wt) fat diet for 1 yr. At the end of 12 mo, islet amyloid deposition was quantified from thioflavin S-stained pancreas sections. The majority of mice in all groups developed islet amyloid (BL6: 91%, F(1): 76%, DBA2: 100%). However, the prevalence (%amyloid-positive islets; BL6: 14 +/- 3%, F(1): 44 +/- 8%, DBA2: 49 +/- 9%, P < 0.05) and severity (%islet area occupied by amyloid; BL6: 0.03 +/- 0.01%, F(1): 9.2 +/- 2.9%, DBA2: 5.7 +/- 2.3%, p < or = 0.01) were significantly lower in BL6 than F(1) and DBA2 mice. Increased islet amyloid severity was negatively correlated with insulin-positive area per islet, in F(1) (r(2) = 0.75, P < 0.001) and DBA2 (r(2) = 0.87, P < 0.001) mice but not BL6 mice (r(2) = 0.07). In summary, the extent of islet amyloid formation in hIAPP transgenic mice is determined by background strain, with mice expressing DBA/2J genes (F(1) and DBA2 mice) being more susceptible to amyloid deposition that replaces beta-cell mass. These findings underscore the importance of genetic and environmental factors in studying metabolic disease. PMID- 15899942 TI - Interleukin-6 is a negative regulator of visfatin gene expression in 3T3-L1 adipocytes. AB - Visfatin is a novel adipocytokine exerting insulin-mimetic effects in various insulin-sensitive tissues such as liver, muscle, and fat. In contrast, interleukin (IL)-6 is a proinflammatory adipose-secreted factor that induces insulin resistance and plasma concentrations that correlate with the development of type 2 diabetes mellitus. In the present study, the impact of IL-6 on visfatin gene expression in 3T3-L1 adipocytes was determined by quantitative real-time reverse transcription-polymerase chain reaction. Interestingly, 30 ng/ml IL-6 time-dependently downregulated visfatin synthesis with a significant 40% suppression seen after 4 h of treatment. Furthermore, the addition of IL-6 for 16 h dose-dependently suppressed visfatin mRNA with significant effects first observed at concentrations as low as 3 ng/ml and a maximal 43% reduction at 30 ng/ml effector. Moreover, inhibitor studies suggested that the negative effect of IL-6 on visfatin expression is, at least in part, mediated by p44/42 mitogen activated protein kinase. In contrast, troglitazone did not reverse the negative effect of IL-6 on visfatin synthesis under these conditions. Taken together, our study suggests that IL-6 might influence glucose tolerance in part by regulation of the novel insulin-mimetic adipocytokine visfatin. PMID- 15899943 TI - Desensitization of the JAK2/STAT5 GH signaling pathway associated with increased CIS protein content in liver of pregnant mice. AB - Chronic exposure to growth hormone (GH) was related to the desensitization of the JAK2/STAT5 signaling pathway in liver, as demonstrated in cells, female rats, and transgenic mice overexpressing GH. The cytokine-induced suppressor (CIS) is considered a major mediator of this desensitization. Pregnancy is accompanied by an increment in GH circulating levels, which were reported to be associated with hepatic GH resistance, although the molecular mechanisms involved in this resistance are not clearly elucidated. We thus evaluated the JAK2/STAT5b signaling pathway and its regulation by the suppressors of cytokine signaling (SOCS)/CIS family and the JAK2-interacting protein SH2-Bbeta in pregnant mouse liver, a model with physiological prolonged exposure to high GH levels. Basal tyrosyl phosphorylation levels of JAK2 and STAT5b in pregnant mice were similar to values obtained for virgin animals, in spite of the important increment of GH they exhibit. Moreover, these signaling mediators were not phosphorylated upon GH stimulation in pregnant mice. A 3.3-fold increase of CIS protein content was found for pregnant mice, whereas the abundance of the other SOCS proteins analyzed and SH2-Bbeta did not significantly change compared with virgin animals. The desensitization of the JAK2/STAT5b GH signaling pathway observed in pregnant mice would then be mainly related to increased CIS levels rather than to the other regulatory proteins examined. PMID- 15899944 TI - Gene expression analysis of Tek/Tie2 signaling. AB - The elaboration of the vasculature during embryonic development involves restructuring of the early vessels into a more complex vascular network. Of particular importance to this vascular remodeling process is the requirement of the Tek/Tie2 receptor tyrosine kinase. Mouse gene-targeting studies have shown that the Tie2-deficient embryos succumb to embryonic death at midgestation due to insufficient sprouting and remodeling of the primary capillary plexus. To identify the underlying genetic mechanisms regulating the process of vascular remodeling, transcriptomes modulated by Tie2 signaling were analyzed utilizing serial analysis of gene expression (SAGE). Two libraries were constructed and sequenced using embryonic day 8.5 yolk sac tissues from Tie2 wild-type and the Tie2-null littermates. After tag extraction, 45,689 and 45,275 SAGE tags were obtained for the Tie2 wild-type and Tie2-null libraries, respectively, yielding a total of 21,376 distinct tags. Close to 62% of the tags were uniquely annotated, whereas 10% of the total tags were unknown. Using semiquantitative PCR, the differential expression of eight genes was confirmed that included Elk3, an important angiogenic switch gene which was upregulated in the absence of Tie2 signaling. The results of this study provide valuable insight into the potential association between Tie2 signaling and other known angiogenic pathways as well as genes that might have novel functions in vascular remodeling. PMID- 15899945 TI - Functional characterization of ACDP2 (ancient conserved domain protein), a divalent metal transporter. AB - We have begun to identify and characterize genes that are differentially expressed with low magnesium. One of these sequences conformed to the ancient conserved domain protein, ACDP2. Real-time RT-PCR of mRNA isolated from distal epithelial cells cultured in low-magnesium media relative to normal media and in kidney cortex of mice maintained on low-magnesium diets compared with those animals consuming normal diets confirmed that the ACDP2 transcript is responsive to magnesium. Mouse ACDP2 was cloned from mouse distal convoluted tubule cells, expressed in Xenopus laevis oocytes, and studied with two-electrode voltage-clamp studies. When expressed in oocytes, ACDP2 mediates saturable Mg2+ uptake with a Michaelis constant of 0.56 +/- 0.05 mM. Transport of Mg2+ by ACDP2 is rheogenic, is voltage-dependent, and is not coupled to Na+ or Cl- ions. Expressed ACDP2 transports a range of divalent cations: Mg2+, Co2+, Mn2+, Sr2+, Ba2+, Cu2+, and Fe2+; accordingly, it is a divalent cation transporter with wide substrate selectivity. The cations Ca2+, Cd2+, Zn2+, and Ni2+ did not induce currents, and only Zn2+ effectively inhibited transport. The ACDP2 transcript is abundantly present in kidney, brain, and heart with lower amounts in liver, small intestine, and colon. Moreover, ACDP2 mRNA is upregulated with magnesium deficiency, particularly in the distal convoluted tubule cells, kidney, heart, and brain. These studies suggest that ACDP2 may provide a regulated transporter for Mg2+ and other divalent cations in epithelial cells. PMID- 15899946 TI - Mitogenic effects of the up-regulation of minichromosome maintenance proteins in anaplastic thyroid carcinoma. AB - CONTEXT: Anaplastic thyroid carcinomas (ATC) are among the most aggressive human malignancies and are characterized by high mitotic activity. Minichromosome maintenance proteins (MCM) 2-7 are required to initiate eukaryotic DNA replication, and their overexpression has been associated with dysplasia and malignancy. OBJECTIVE: In an attempt to cast light on the mechanisms governing ATC, we evaluated MCM5 and MCM7 expression in human normal, papillary (PTC), and anaplastic thyroid samples, as well as in primary culture cells and transgenic mouse models. RESULTS: MCM5 and MCM7 expression was high in 65% of ATC and negligible in normal thyroid tissue and papillary thyroid carcinomas. In ATC, high MCM5 and MCM7 expression was paralleled by high levels of MCM2 and MCM6. An analysis of human ATC primary cell cultures and of a transgenic mouse model of ATC confirmed these findings. An increased transcription rate accounted for MCM7 up-regulation, because the activity of the MCM7 promoter was more than 10-fold higher in ATC cells compared with normal thyroid cells. Adoptive overexpression of wild-type p53, but not of its inactive (R248W and R273H) mutants, strongly down-regulated transcription from the MCM7 promoter, suggesting that p53 knock out contributes to MCM7 up-regulation in ATC. Treatment with small inhibitory duplex RNAs, which decrease MCM7 protein levels, reduced the rate of DNA synthesis in ATC cells. CONCLUSION: MCM proteins are overexpressed in ATC and sustain the high proliferative capacity of ATC cells. PMID- 15899947 TI - Increased transvascular lipoprotein transport in diabetes: association with albuminuria and systolic hypertension. AB - CONTEXT: Diabetes is associated with a highly increased risk of atherosclerosis, especially if hypertension or albuminuria is present. OBJECTIVE: We hypothesized that the increased transvascular lipoprotein transport in diabetes may be further accelerated if hypertension or albuminuria is present, possibly explaining increased intimal lipoprotein accumulation and thus atherosclerosis. DESIGN: The study was cross-sectional and was performed in 1999-2002. SETTING: The study took place in the referral center. PATIENTS: The patients included 60 with diabetes mellitus (27 with type 1 diabetes and 33 with type 2 diabetes) and 42 healthy controls. All were randomly recruited. MAIN OUTCOME MEASURE: We used an in vivo method for measurement of transvascular transport of low-density lipoprotein (LDL). Autologous 131I-LDL was reinjected iv, and the 1-h fractional escape rate was taken as an index of transvascular transport. RESULTS: Transvascular LDL transport was 1.8 (1.6-2.0), 2.3 (2.0-2.6), and 2.6 (1.3-4.0)%/[h x (liter/m2)] in healthy controls, diabetic controls, and diabetes patients with systolic hypertension or albuminuria, respectively (P = 0.013; F = 4.5; df =2; ANOVA). These differences most likely were not caused by altered hepatic LDL receptor expression, glycosylation of LDL, small LDL size, or medicine use. CONCLUSIONS: Transvascular LDL transport is increased in patients with diabetes mellitus, especially if systolic hypertension or albuminuria is present. Accordingly, lipoprotein flux into the arterial wall could be increased in these patients, possibly explaining accelerated development of atherosclerosis. PMID- 15899948 TI - Association between a protein polymorphism in the start codon of the vitamin D receptor gene and severe diabetic retinopathy in C-peptide-negative type 1 diabetes. AB - CONTEXT: The vitamin D (VD) receptor (VDR) is extensively expressed in retina. The plasma concentration of 1,25-dihydroxyvitamin D3 has been inversely correlated with the severity of diabetic retinopathy (DR), which raises the possibility that VD, through its antiinflammatory, antioxidant, antiproliferative, and antiangiogenic properties, may protect diabetic retina. The TaqI VDR polymorphism has been associated with severe DR. The FokI VDR polymorphism is a T-to-C substitution in the first codon (f allele), abolishing the first translation initiation site and resulting in a peptide lacking three amino acids (F allele), which increases the transcriptional activity of VDR. OBJECTIVE AND DESIGN: To examine whether FokI polymorphism is involved in severe DR, 254 Caucasians with longstanding C-peptide-negative type 1 diabetes, 128 patients with absent/mild DR (control group), and 126 patients with preproliferative/proliferative DR (study group) were genotyped using PCR restriction fragment length polymorphism analysis. RESULTS: The genotype distribution was in Hardy-Weinberg equilibrium and was different between groups (P = 0.046). The frequency of F allele was significantly higher in the control (66.4%) than in the study group (56%, odds ratio = 0.64, 95% confidence interval 0.44-0.92, P = 0.016). In subjects with fewer than 25 yr of diabetes duration (median value, n = 134), this association was strongly increased (P = 0.0008). CONCLUSIONS: In conclusion, we observed, in a cohort of Caucasians with C-peptide negative type 1 diabetes, a novel association between the functional FokI VDR polymorphism and severe DR, especially among subjects with fewer than 25 yr of diabetes duration. PMID- 15899949 TI - Dyslipidemia and metabolic syndrome in the sisters of women with polycystic ovary syndrome. AB - CONTEXT: Dyslipidemia is a feature of polycystic ovary syndrome (PCOS), but its pathogenesis remains controversial. OBJECTIVE: The objective of this study was to test the hypothesis that dyslipidemia is a heritable trait in sisters of women with PCOS. DESIGN: A case-control design was used. SETTING: The study took place at General Clinical Research Centers in four academic medical centers in the United States. PATIENTS: The subjects included 385 sisters of women with PCOS with the following reproductive phenotypes: sisters with PCOS (n = 51), sisters with hyperandrogenemia and regular menses (HA) (n = 38), unaffected sisters (n = 143), and unknown phenotypes (n = 153). One hundred twenty-five control women of comparable age, body mass index, and ethnicity to women with PCOS were included. INTERVENTIONS: Fasting blood was obtained for measurements of lipid profile, reproductive hormones, glucose, and insulin levels. MAIN OUTCOME MEASURES: The main outcome measures included lipid and lipoprotein levels and prevalence of metabolic syndrome. RESULTS: Sisters with PCOS and HA phenotypes had higher total (P < or = 0.001) and low-density lipoprotein cholesterol levels (P < or = 0.01) compared with unaffected sisters and control women. Triglyceride levels were elevated only in sisters with the PCOS phenotype (P < 0.05). The prevalence of metabolic syndrome was increased in sisters with the PCOS (n = 29) and HA (n = 17) phenotypes compared with unaffected sisters (n = 85) (P < 0.001 and P < 0.05, respectively). CONCLUSIONS: Low-density lipoprotein levels are increased in affected sisters of women with PCOS consistent with a heritable trait. The prevalence of metabolic syndrome is increased in affected sisters. PMID- 15899950 TI - L-folic acid supplementation in healthy postmenopausal women: effect on homocysteine and glycolipid metabolism. AB - CONTEXT: Hyperhomocysteinemia as well as alterations of glycemic and lipidic metabolism are recognized as risk factors for cardiovascular diseases. OBJECTIVE: The aim of this study was to examine the effect of L-folic acid supplementation on homocysteine (Hcy) and related thiols, such as cysteine (Cys) and Cys-glycine (Cys-Glyc) pathways and their relationship to glucose, insulin, and lipidic metabolism in normoinsulinemic postmenopausal women. DESIGN: This study was a randomized placebo, not double-blind, trial. SETTING: The study was performed in an academic research center. PATIENTS OR OTHER PARTICIPANTS: Twenty healthy postmenopausal women were selected. No patient was taking drugs known to affect lipid or glucose metabolism. INTERVENTION(S): Patients underwent two hospitalizations before and after 8 wk of L-acid folic (7.5 mg/d) or placebo administration. The glycemic metabolism was studied by an oral glucose tolerance test and a hyperinsulinemic euglycemic clamp. Hcy metabolism was studied by a standardized oral methionine-loading test. MAIN OUTCOME MEASURE(S): Hcy, Cys, and Cys-Glyc, basally and after a methionine loading test, were measured. Basal insulin, glucose, and peptide C levels as well as area under the curve for insulin, area under the curve for peptide, hepatic insulin extraction, and metabolic index were assayed. The total cholesterol, high-density lipoprotein (HDL) cholesterol, and low-density lipoprotein (LDL) cholesterol levels and the cholesterol/HDL and LDL/HDL ratios were also measured. RESULTS: The total basal Hcy concentration and the plasma postmethionine loading Hcy values were significantly decreased (P < 0.01) in L-folic acid-treated patients, whereas postmethionine loading Cys-Glyc levels were markedly increased (P < 0.02). Furthermore, L-folic acid intake induced a significant improvement in carbohydrate metabolism through an increase in fractional hepatic insulin extraction (P < 0.05) and peripheral insulin sensitivity (P < 0.02) in normoinsulinemic women. HDL levels considerably increased, inducing an improvement in other atherosclerotic indexes, such as cholesterol/HDL and LDL/HDL ratios (P < 0.03). CONCLUSIONS: These results show that folic acid supplementation lowers plasma Hcy levels and improves insulin and lipid metabolism, reducing the risk of cardiovascular disease. PMID- 15899951 TI - The relationship between parity and bone mineral density in women characterized by a homogeneous lifestyle and high parity. AB - CONTEXT: We reported previously that Old Order Amish (OOA) women have fewer hip fractures and higher bone mineral density (BMD) than non-Amish Caucasian women. OBJECTIVE: The objective of this study was to determine whether the high parity characteristic of OOA women contributes to their relative bone health. Previous data on the long-term effects of parity on BMD have yielded conflicting results with few data from very high parity populations. This observational study included participants in the Amish Family Osteoporosis Study, begun in 1997 to identify genetic and clinical determinants of osteoporosis in the OOA. We measured BMD by dual-energy x-ray absorptiometry at the spine, hip, and distal radius in 424 parous OOA women aged 40 and older (mean age, 57.7 +/- 12 yr; mean parity, 7.6 +/- 2.9). RESULTS: Increasing parity was associated with later menopause (P = 0.001) and modestly, but not significantly, higher body mass index (BMI) (P = 0.09). Increasing parity was associated with higher BMD at the total hip and trochanter (age-adjusted P = 0.02 and 0.03), no longer statistically significant after accounting for BMI. Among women aged 50-59 yr, parity was strongly associated with BMD even after accounting for age and BMI (age-adjusted P = 0.02), although this was not true for women younger than 50 or at least 60 yr old. CONCLUSIONS: We conclude that high parity is associated with increased hip BMD in OOA women, largely mediated by higher BMI. The parity-hip BMD association remained statistically significant after accounting for age and BMI only in women aged 50-59 yr, partially explained by a later menopausal age with high parity. The benefit of high parity on BMD appeared to be lost soon after the menopausal transition, and, therefore, these data provide evidence of neither a detrimental nor beneficial effect of high parity on long-term bone health. PMID- 15899952 TI - Safety of growth hormone treatment in pediatric patients with idiopathic short stature. AB - CONTEXT: Recombinant human GH was approved by the United States Food and Drug Administration in 2003 for the treatment of idiopathic short stature (ISS). However, to date, the safety of GH in this patient population has not been rigorously studied. OBJECTIVE: The objective of this study was to address the safety of GH treatment in children with ISS compared with GH safety in patient populations for which GH has been approved previously: Turner syndrome (TS) and GH deficiency (GHD). DESIGN/SETTING: The rates of serious adverse events (SAEs) and adverse events (AEs) of particular relevance to GH-treated populations were compared across the three patient populations among five multicenter GH registration studies. PATIENTS: Children with ISS, TS, or GHD were studied. INTERVENTION: Treatment consisted of GH doses ranging from 0.18-0.37 mg/kg.wk. MAIN OUTCOME MEASURES: The main outcome measures were rates of SAEs and AEs of special relevance to patients receiving GH. Laboratory measures of carbohydrate metabolism were used as outcome measures for the ISS studies. RESULTS: Within the ISS studies, comprising one double-blind, placebo-controlled study and one open label, dose-response study, SAEs (mainly hospitalizations for accidental injury or acute illness unrelated to GH exposure) were reported for 13-14% of GH-treated patients. Overall AE rates (serious and nonserious) as well as rates of potentially GH-related AEs were similar in the GHD, TS, and ISS studies (for ISS studies combined: otitis media, 8%; scoliosis, 3%; hypothyroidism, 0.7%; changes in carbohydrate metabolism, 0.7%; hypertension, 0.4%). Measures of carbohydrate metabolism were not affected by GH treatment in patients with ISS. There was no significant GH effect on fasting blood glucose in either study (GH dose range, 0.22-0.37 mg/kg.wk) or on insulin sensitivity (placebo-controlled study only). CONCLUSION: GH appears safe in ISS; however, the studies were not powered to assess the frequency of rare GH-related events, and longer-term follow-up studies of GH-treated patients with ISS are warranted. PMID- 15899953 TI - CLINICAL REVIEW: Use of antiepileptic drugs in the treatment of chronic painful diabetic neuropathy. AB - CONTEXT: Up to 25% of individuals with diabetes develop painful diabetic neuropathy, suffering spontaneous pain, allodynia, hyperalgesia, and other unpleasant symptoms. Decreased physical activity, increased fatigue, and mood and sleep problems may result. EVIDENCE ACQUISITION: A MEDLINE search was conducted, limiting searching to double-blind, randomized, controlled trials (1978 to present) of antiepileptic drugs (carbamazepine, gabapentin, pregabalin, topiramate, and lamotrigine) used in the treatment of chronic neuropathic pain. EVIDENCE SYNTHESIS: The most important aspect of treatment is targeted at modification of the underlying disease. However, approaches to symptomatic pain control are essential and include multiple drug classes. Tricyclic antidepressants, including imipramine, nortriptyline, and amitriptyline, have been the mainstays of treatment, but anticholinergic effects, such as dry mouth, blurring of vision, constipation, orthostatic hypotension, and cardiac arrhythmias, as well as other adverse effects, often limit their use. Other treatments include capsaicin, clonidine, acupuncture, and electrical stimulation, suggesting that there is no single effective treatment. First-generation antiepileptic drugs have been shown to be effective in neuropathic pain. The evidence supporting the use of a new generation of antiepileptic drugs in painful diabetic neuropathy is reviewed. PMID- 15899954 TI - Height and height Z-score are related to calcium absorption in five- to fifteen year-old girls. AB - CONTEXT: Understanding the relationship between calcium absorption and growth has been limited. We have developed a database of calcium absorption measurements in 315 girls aged 5.0-15.0 yr. DESIGN: We have used this database to assess the relationship between height, its age- and gender-normalized value (height Z score), and calcium absorptive efficiency. RESULTS: Overall, height was significantly related to calcium absorption (corrected for calcium intake, age, Tanner, stage, and ethnicity) (P = 0.001). Similarly, height Z-score was significantly related to calcium absorption (P < 0.007). About 3-3.5% of the variability (eta2) of absorption was associated with height or height Z-score. We found that calcium absorption was significantly lower in girls with height Z score equal to or less than zero compared with those with a height Z-score more than zero (difference, 3.9 +/- 1.4%, mean +/- sem; P = 0.007). Limiting the analysis to those girls in which Tanner staging was performed or those age 9 yr or older did not substantively affect these relationships. CONCLUSION: These results indicate that a small but significant component of the variability in calcium absorption is due to height. Identifying genetic risk factors for lowered calcium absorption during growth could lead to individual approaches for prevention of inadequate bone mass. PMID- 15899955 TI - Insulin-induced hypoglycemia accelerates gastric emptying of solids and liquids in long-standing type 1 diabetes. AB - CONTEXT: The rate of gastric emptying of carbohydrate is a major determinant of postprandial glycemia. In healthy subjects and patients with uncomplicated type 1 diabetes, there is evidence that gastric emptying may be accelerated by insulin induced hypoglycemia. OBJECTIVE: The objective was to determine the effects of acute hypoglycemia on gastric emptying in long-standing type 1 diabetes and evaluate whether the response to hypoglycemia is influenced by the rate of gastric emptying during euglycemia and/or autonomic dysfunction. DESIGN: Gastric emptying of a solid/liquid meal (100 g (99m)Tc-minced beef and 150 ml 67Ga-EDTA labeled water) was measured by scintigraphy on 2 separate days, during hypoglycemia and euglycemia. SETTING: These studies took place at the Department of Nuclear Medicine, Positron Emission Tomography, and Bone Densitometry at the Royal Adelaide Hospital. PATIENTS: Twenty type 1 patients (4 female, 16 male; age, 45.9 +/- 2.3 yr; duration of known diabetes, 18.0 +/- 2.7 yr) were recruited from outpatient clinics and the Diabetes Centre at the Royal Adelaide Hospital. INTERVENTION: Hypoglycemia (approximately 2.6 mmol/liter) was established 15 min before and maintained for 45 min after meal consumption. On one of the days, autonomic nerve function was evaluated using cardiovascular reflex tests. MAIN OUTCOME MEASURE: The main outcome measure was gastric emptying during hypoglycemia when compared with euglycemia. RESULTS: Twelve of the 20 subjects had autonomic neuropathy. Gastric emptying of both solid (P < 0.001) and liquid (P < 0.05) was faster during hypoglycemia. The magnitude of this acceleration was greater when the rate of gastric emptying during euglycemia was slower (solid, percentage retention at 100 min, r = -0.52, P < 0.05; liquid, 50% emptying time, r = -0.82, P < 0.0001, but not influenced by autonomic nerve function). CONCLUSIONS: Insulin-induced hypoglycemia accelerates gastric emptying of solids and liquids in long-standing type 1 diabetes, even in those patients with delayed emptying, and is likely to be an important mechanism in the counter-regulation of hypoglycemia. PMID- 15899956 TI - Quality of life in childhood onset growth hormone-deficient patients in the transition phase from childhood to adulthood. AB - BACKGROUND: Quality of life (QoL) has not been specifically assessed in GH deficient (GHD) transition patients. METHODS: We assessed QoL at baseline and after 1 and 2 yr of GH treatment in severely GHD patients, using an adult GHD specific questionnaire, QLS(M)-H. Subjects were randomized to GH, 25.0 microg/kg.d (n = 25) or 12.5 microg/kg.d (n = 28), or no treatment (n = 13). sd scores for QLS-H were calculated from normative data, specific to country of origin, gender, and age range of the patients. RESULTS: Baseline QLS-H sd scores were -0.35 +/- 1.17 in females and -0.70 +/- 1.05 in males (P = 0.280). sd scores for individual dimensions of ability to become sexually aroused, ability to tolerate stress, body shape, concentration, initiative/drive, physical stamina, and self-confidence were significantly lower than the normal average. Particularly affected were body shape (sd score, -0.80 +/- 0.99; quartile (Q)1:Q3, -1.52:-0.29) and sexual arousal (sd score, -0.41 +/- 0.88; Q1:Q3, -1.15: 0.13). Total QLS-H sd score increased slightly but not significantly for combined GH treatment groups compared with control at yr 1 (0.047 +/- 1.51 vs. -0.32 +/- 1.66; P = 0.845) but not after yr 2 (-0.00 +/- 0.80 vs. 0.12 +/- 0.89; P = 0.385); no dose effect of GH was observed. GH treatment significantly increased sd score from baseline to yr 2 for sexual arousal and body shape (0.23 +/- 0.78, P = 0.038; and 0.46 +/- 1.26, P = 0.035, respectively). CONCLUSION: Although overall baseline QoL was not compromised in severely GHD patients during the transition period, dimensions related to age-specific psychological problems were significantly worse than healthy subjects and appeared to positively respond to GH treatment. PMID- 15899957 TI - Normal secretion and action of the gut incretin hormones glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide in young men with low birth weight. AB - CONTEXT: Low birth weight (LBW) is associated with increased risk of type 2 diabetes mellitus. An impaired incretin effect was reported previously in type 2 diabetic patients. OBJECTIVE: We studied the secretion and action of glucagon like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) in young LBW men (n = 24) and matched normal birth weight controls (NBW) (n = 25). RESULTS: LBW subjects were 5 cm shorter but had a body mass index similar to NBW. LBW subjects had significantly elevated fasting and postprandial plasma glucose, as well as postprandial (standard meal test) plasma insulin and C-peptide concentrations, suggestive of insulin resistance. Insulin secretion in response to changes in glucose concentration ("beta-cell responsiveness") during the meal test was similar in LBW and NBW but inappropriate in LBW relative to insulin sensitivity. Fasting and postprandial plasma GLP-1 and GIP levels were similar in the groups. First- and second-phase insulin responses were similar in LBW and NBW during a hyperglycemic clamp (7 mm) with infusion of GLP-1 or GIP, respectively, demonstrating normal action of these hormones on insulin secretion. CONCLUSION: Reduced secretion or action of GLP-1 or GIP does not explain a relative reduced beta-cell responsiveness to glucose or the slightly elevated plasma glucose concentrations observed in young LBW men. PMID- 15899958 TI - Outcome of gamma knife radiosurgery in 82 patients with acromegaly: correlation with initial hypersecretion. AB - CONTEXT: Because surgical and medical therapies of acromegaly all have specific limitations, radiotherapy has been used as an adjunctive strategy. Stereotactic radiosurgery has not yet been widely evaluated. OBJECTIVE: The objective was to perform an analysis of long-term hormonal effects and tolerance of gamma knife radiosurgery. DESIGN: Eighty-two patients were prospectively studied over a decade, with a mean follow-up of 49.5 months. SETTING: All patients were treated at the Department of Functional Neurosurgery of Marseille, France. PATIENTS: The patients included 82 with active acromegaly, of whom 63 had previous transsphenoidal surgery. INTERVENTION: Intervention included radiosurgery using the Leksell Gamma Unit B model. MAIN OUTCOME MEASURES: Remission was diagnosed when mean GH levels were less than 2 ng/ml and IGF-I was normal for age off somatostatin agonists (at least 3 months). RESULTS: Seventeen percent of the patients were in remission without any treatment. Twenty-three percent previously uncontrolled on somatostatin agonists fulfilled the same criteria after gamma knife while maintained on medical treatment. Initial GH and IGF-I levels off somatostatin agonists were significantly higher in uncured than in remission group (P = 0.01 and 0.047, respectively). Withdrawal of somatostatin agonists at the time of radiosurgery had no incidence on the outcome. No significant difference was found in success rate whether patients had previously been treated or not. Long-term side effects included complete (n = 2) or partial (n = 12) hypopituitarism diagnosed 1-7 yr after gamma knife. CONCLUSIONS: Gamma knife radiosurgery may represent a therapeutic approach in patients with moderate initial or residual GH hypersecretion. PMID- 15899959 TI - Vitamin D insufficiency does not affect bone mineral density response to raloxifene. AB - CONTEXT: Vitamin D insufficiency and osteoporosis are common and often coexist in postmenopausal women. OBJECTIVE: The objective of this study was to test whether the presence of vitamin D insufficiency at the initiation of raloxifene therapy affected the subsequent response of bone mineral density (BMD). DESIGN, SETTING, AND PARTICIPANTS: We studied 7522 postmenopausal participants of the Multiple Outcomes of Raloxifene Evaluation, a placebo-controlled trial of the effects of raloxifene on BMD and fracture. INTERVENTION: After enrollment, all participants began daily supplements of 500 mg calcium and 400-600 IU cholecalciferol; 1 month later, women were randomly assigned to placebo or raloxifene. MAIN OUTCOME MEASURE: Serum levels of vitamin D [25-hydroxy vitamin D (25OHD)] were measured at enrollment, randomization, and 6 months later. We categorized participants' vitamin D status (deficient, insufficient, or sufficient) based on their randomization 25OHD level. We estimated the effects of treatment on BMD within these subgroups using linear regression models. RESULTS: At enrollment, 3.2% of participants were vitamin D deficient, and 51.8% were insufficient; after 7 months of cholecalciferol supplementation, 0.2% of all participants remained D deficient, and 23.6% remained insufficient. The effects of raloxifene on hip and spine BMD did not vary by vitamin D status at randomization (P = 0.08 and P = 0.7, respectively). CONCLUSION: We conclude that vitamin D status at initiation of raloxifene therapy does not affect the subsequent BMD response when coadministered with cholecalciferol and calcium. After 7 months of cholecalciferol therapy, very few women continued to have 25OHD levels in the deficient range; however, 25OHD levels remained suboptimal in nearly one fourth of the cohort. Additional research is needed to determine whether these observations can be generalized to other antiresorptive agents. PMID- 15899960 TI - Association of cryptorchidism with a specific haplotype of the estrogen receptor alpha gene: implication for the susceptibility to estrogenic environmental endocrine disruptors. AB - CONTEXT: The prevalence of cryptorchidism (CO) has increased during the past few decades in several countries, and this event has primarily been ascribed to the estrogenic effects of environmental endocrine disruptors (EEDs). Little is known, however, about the role of genetic susceptibility to EEDs in this phenomenon. OBJECTIVE: The objective of this study was to determine whether CO is associated with a specific haplotype of the gene for estrogen receptor alpha (ESR1) that mediates the estrogenic effects of EEDs. DESIGN: This was a case-control study. SETTING: The study was performed at the National Research Institute and University Hospitals. SUBJECTS: Sixty-three cryptorchid males, aged 1-13 yr, and 47 control males, aged 4-12 yr, were studied. INTERVENTION: After genotyping 15 single nucleotide polymorphisms widely distributed in the greater than 300-kb genomic sequences of ESR1, haplotype analysis was performed. MAIN OUTCOME MEASURE: Identification of a specific ESR1 haplotype associated with CO was the main outcome measure. RESULTS: A haplotype block was identified for an approximately 50-kb region encompassing single nucleotide polymorphisms 10-14 in the 3' region of ESR1 in both groups. The frequency of the estimated AGATA haplotype within the block was higher in the patients than in the control males (34.0% vs. 21.3%; P = 0.037), and the association of this haplotype with CO phenotype was significant in a recessive mode (P = 0.0060). The homozygosity for this haplotype was identified only in the patients, and the frequency of the homozygotes was significantly different between the two groups (10 of 63 vs. zero of 47; P = 0.0042). CONCLUSIONS: The association of CO with homozygosity for the specific ESR1 haplotype suggests the relevance of genetic susceptibility to EEDs in the development of CO. PMID- 15899961 TI - Defining the proinflammatory phenotype using high sensitive C-reactive protein levels as the biomarker. AB - CONTEXT: Inflammation is pivotal in atherosclerosis. The prototypic marker of inflammation is C-reactive protein (CRP). Numerous studies have confirmed that high CRP levels in normal volunteers predict cardiovascular events. OBJECTIVE: The objective of this study was to define proximal and associated abnormalities of the proinflammatory phenotype using CRP levels as the biomarker. DESIGN AND SUBJECTS: Two groups of normal, healthy subjects, selected by stringent criteria from an initial cohort of 252, were studied over the period of 12 months. Group 1 included subjects with consistently low CRP (<0.004 microM or <0.5 mg/liter; low CRP group; n = 15). Group 2 included subjects with consistently high CRP (>2.0 or >0.016 microM to <10 mg/liter or <0.085 microM; high CRP group; n = 13). MAIN OUTCOME MEASURES: Fasting blood (50 ml) was obtained, and the following parameters were assayed: high sensitivity CRP, fibrinogen, lipid profile, insulin, whole blood cytokines after stimulation with lipopolysaccharide (LPS; 100 ng/ml for 24 h), soluble cell adhesion molecules, plasminogen activator inhibitor-1, CD40, CD40 ligand, leptin, adiponectin, monocyte chemoattractant protein-1, IL-8, matrix metalloproteinase-3 (MMP-3), and MMP-9. Genomic DNA was obtained from peripheral blood leukocytes, and the TNF-alpha -308 genotype was determined. RESULTS: The median CRP levels were 0.0018 microM (0.21 mg/liter) and 0.031 microM (3.7 mg/liter) for the low and high groups, respectively. High CRP subjects were older and had significantly higher body mass indexes, triglycerides, insulin, homeostasis model assessment, and leptin levels compared with low CRP subjects. The markers of inflammation, plasminogen activator inhibitor-1, MMP-9, fibrinogen, and vascular cell adhesion molecule-1 levels were significantly higher in the high compared with the low CRP group. LPS-stimulated levels of whole blood IL-1beta, IL-6, and TNF were significantly higher, and IL-4 levels were significantly lower in the high CRP group. After age- and body mass index-adjusted analysis of covariance, only plasma MMP-9 levels and LPS stimulated whole blood IL-1beta and TNF levels were significantly higher in the high CRP group. The frequency of the rare A allele at TNF-alpha -308 was equivalent in high and low CRP groups. CONCLUSIONS: A phenotype characterized by increased plasma inflammatory mediators as well as increased LPS-stimulated whole blood TNF-alpha and IL-1beta levels is associated with high plasma CRP levels. This systemic inflammatory phenotype may contribute to vascular inflammation or may reflect inflammation in vessels or at other sites. PMID- 15899962 TI - Are California's large employers moving to catastrophic health insurance coverage? AB - Large employers in California are experimenting with new health benefit and insurance options as premium rates continue to escalate. This study examines the offer and penetration rates of catastrophic coverage insurance products, including high-deductible PPO and consumer-driven health plans, among large California employers before the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 was passed. Only a few employers offered these plans, and they did not provide adequate incentives for their workers to accept. California employers, like the rest of the nation, implemented these plans as alternatives rather than replacements to their current plan choices. PMID- 15899963 TI - Comparative and functional genomic analyses of the pathogenicity of phytopathogen Xanthomonas campestris pv. campestris. AB - Xanthomonas campestris pathovar campestris (Xcc) is the causative agent of crucifer black rot disease, which causes severe losses in agricultural yield world-wide. This bacterium is a model organism for studying plant-bacteria interactions. We sequenced the complete genome of Xcc 8004 (5,148,708 bp), which is highly conserved relative to that of Xcc ATCC 33913. Comparative genomics analysis indicated that, in addition to a significant genomic-scale rearrangement cross the replication axis between two IS1478 elements, loss and acquisition of blocks of genes, rather than point mutations, constitute the main genetic variation between the two Xcc strains. Screening of a high-density transposon insertional mutant library (16,512 clones) of Xcc 8004 against a host plant (Brassica oleraceae) identified 75 nonredundant, single-copy insertions in protein-coding sequences (CDSs) and intergenic regions. In addition to known virulence factors, full virulence was found to require several additional metabolic pathways and regulatory systems, such as fatty acid degradation, type IV secretion system, cell signaling, and amino acids and nucleotide metabolism. Among the identified pathogenicity-related genes, three of unknown function were found in Xcc 8004-specific chromosomal segments, revealing a direct correlation between genomic dynamics and Xcc virulence. The present combination of comparative and functional genomic analyses provides valuable information about the genetic basis of Xcc pathogenicity, which may offer novel insight toward the development of efficient methods for prevention of this important plant disease. PMID- 15899964 TI - Direct isolation and identification of promoters in the human genome. AB - Transcriptional regulatory elements play essential roles in gene expression during animal development and cellular response to environmental signals, but our knowledge of these regions in the human genome is limited despite the availability of the complete genome sequence. Promoters mark the start of every transcript and are an important class of regulatory elements. A large, complex protein structure known as the pre-initiation complex (PIC) is assembled on all active promoters, and the presence of these proteins distinguishes promoters from other sequences in the genome. Using components of the PIC as tags, we isolated promoters directly from human cells as protein-DNA complexes and identified the resulting DNA sequences using genomic tiling microarrays. Our experiments in four human cell lines uncovered 252 PIC-binding sites in 44 semirandomly selected human genomic regions comprising 1% (30 megabase pairs) of the human genome. Nearly 72% of the identified fragments overlap or immediately flank 5' ends of known cDNA sequences, while the remainder is found in other genomic regions that likely harbor putative promoters of unannotated transcripts. Indeed, molecular analysis of the RNA isolated from one cell line uncovered transcripts initiated from over half of the putative promoter fragments, and transient transfection assays revealed promoter activity for a significant proportion of fragments when they were fused to a luciferase reporter gene. These results demonstrate the specificity of a genome-wide analysis method for mapping transcriptional regulatory elements and also indicate that a small, yet significant number of human genes remains to be discovered. PMID- 15899965 TI - Ultraconserved elements in insect genomes: a highly conserved intronic sequence implicated in the control of homothorax mRNA splicing. AB - Recently, we identified a large number of ultraconserved (uc) sequences in noncoding regions of human, mouse, and rat genomes that appear to be essential for vertebrate and amniote ontogeny. Here, we used similar methods to identify ultraconserved genomic regions between the insect species Drosophila melanogaster and Drosophila pseudoobscura, as well as the more distantly related Anopheles gambiae. As with vertebrates, ultraconserved sequences in insects appear to occur primarily in intergenic and intronic sequences, and at intron-exon junctions. The sequences are significantly associated with genes encoding developmental regulators and transcription factors, but are less frequent and are smaller in size than in vertebrates. The longest identical, nongapped orthologous match between the three genomes was found within the homothorax (hth) gene. This sequence spans an internal exon-intron junction, with the majority located within the intron, and is predicted to form a highly stable stem-loop RNA structure. Real-time quantitative PCR analysis of different hth splice isoforms and Northern blotting showed that the conserved element is associated with a high incidence of intron retention in hth pre-mRNA, suggesting that the conserved intronic element is critically important in the post-transcriptional regulation of hth expression in Diptera. PMID- 15899966 TI - Identification of genomic features using microsyntenies of domains: domain teams. AB - The detection, across several genomes, of local conservation of gene content and proximity considerably helps the prediction of features of interest, such as gene fusions or physical and functional interactions. Here, we want to process realistic models of chromosomes, in which genes (or genomic segments of several genes) can be duplicated within a chromosome, or be absent from some other chromosome(s). Our approach adopts the technique of temporarily forgetting genes and working directly with protein "domains" such as those found in Pfam. This allows the detection of strings of domains that are conserved in their content, but not necessarily in their order, which we refer to as domain teams. The prominent feature of the method is that it relaxes the rigidity of the orthology criterion and avoids many of the pitfalls of gene-families identification methods, often hampered by multidomain proteins or low levels of sequence similarity. This approach, that allows both inter- and intrachromosomal comparisons, proves to be more sensitive than the classical methods based on pairwise sequence comparisons, particularly in the simultaneous treatment of many species. The automated and fast detection of domain teams, together with its increased sensitivity at identifying segments of identical (protein-coding) gene contents as well as gene fusions, should prove a useful complement to other existing methods. PMID- 15899967 TI - AnoEST: toward A. gambiae functional genomics. AB - Here, we present an analysis of 215,634 EST and cDNA sequences of a major vector of human malaria Anopheles gambiae structured into the AnoEST database. The expressed sequences are grouped into clusters using genomic sequence as template and associated with inferred functional annotation, including the following: corresponding Ensembl gene prediction, putative orthologous genes in other species, homology to known proteins, protein domains, associated Gene Ontology terms, and corresponding classification into broad GO-slim functional groups. AnoEST is a vital resource for interpretation of expression profiles derived using recently developed A. gambiae cDNA microarrays. Using these cDNA microarrays, we have experimentally confirmed the expression of 7961 clusters during mosquito development. Of these, 3100 are not associated with currently predicted genes. Moreover, we found that clusters with confirmed expression are nonbiased with respect to the current gene annotation or homology to known proteins. Consequently, we expect that many as yet unconfirmed clusters are likely to be actual A. gambiae genes. [AnoEST is publicly available at http://komar.embl.de, and is also accessible as a Distributed Annotation Service (DAS).]. PMID- 15899968 TI - Cell-replacement therapy for diabetes: Generating functional insulin-producing tissue from adult human liver cells. AB - Shortage in tissue availability from cadaver donors and the need for life-long immunosuppression severely restrict the large-scale application of cell replacement therapy for diabetic patients. This study suggests the potential use of adult human liver as alternate tissue for autologous beta-cell-replacement therapy. By using pancreatic and duodenal homeobox gene 1 (PDX-1) and soluble factors, we induced a comprehensive developmental shift of adult human liver cells into functional insulin-producing cells. PDX-1-treated human liver cells express insulin, store it in defined granules, and secrete the hormone in a glucose-regulated manner. When transplanted under the renal capsule of diabetic, immunodeficient mice, the cells ameliorated hyperglycemia for prolonged periods of time. Inducing developmental redirection of adult liver offers the potential of a cell-replacement therapy for diabetics by allowing the patient to be the donor of his own insulin-producing tissue. PMID- 15899969 TI - Geometric diffusions as a tool for harmonic analysis and structure definition of data: multiscale methods. AB - In the companion article, a framework for structural multiscale geometric organization of subsets of R(n) and of graphs was introduced. Here, diffusion semigroups are used to generate multiscale analyses in order to organize and represent complex structures. We emphasize the multiscale nature of these problems and build scaling functions of Markov matrices (describing local transitions) that lead to macroscopic descriptions at different scales. The process of iterating or diffusing the Markov matrix is seen as a generalization of some aspects of the Newtonian paradigm, in which local infinitesimal transitions of a system lead to global macroscopic descriptions by integration. This article deals with the construction of fast-order N algorithms for data representation and for homogenization of heterogeneous structures. PMID- 15899970 TI - Geometric diffusions as a tool for harmonic analysis and structure definition of data: diffusion maps. AB - We provide a framework for structural multiscale geometric organization of graphs and subsets of R(n). We use diffusion semigroups to generate multiscale geometries in order to organize and represent complex structures. We show that appropriately selected eigenfunctions or scaling functions of Markov matrices, which describe local transitions, lead to macroscopic descriptions at different scales. The process of iterating or diffusing the Markov matrix is seen as a generalization of some aspects of the Newtonian paradigm, in which local infinitesimal transitions of a system lead to global macroscopic descriptions by integration. We provide a unified view of ideas from data analysis, machine learning, and numerical analysis. PMID- 15899972 TI - The stressed gut: contributions of intestinal stress peptides to inflammation and motility. PMID- 15899973 TI - Vesicle reuse revisited. PMID- 15899974 TI - Biography of Douglas T. Fearon. PMID- 15899975 TI - Human-modified temperatures induce species changes: Joint attribution. AB - Average global surface-air temperature is increasing. Contention exists over relative contributions by natural and anthropogenic forcings. Ecological studies attribute plant and animal changes to observed warming. Until now, temperature species connections have not been statistically attributed directly to anthropogenic climatic change. Using modeled climatic variables and observed species data, which are independent of thermometer records and paleoclimatic proxies, we demonstrate statistically significant "joint attribution," a two-step linkage: human activities contribute significantly to temperature changes and human-changed temperatures are associated with discernible changes in plant and animal traits. Additionally, our analyses provide independent testing of grid-box scale temperature projections from a general circulation model (HadCM3). PMID- 15899976 TI - The rise of the ants: a phylogenetic and ecological explanation. AB - In the past two decades, studies of anatomy, behavior, and, most recently, DNA sequences have clarified the phylogeny of the ants at the subfamily and generic levels. In addition, a rich new harvest of Cretaceous and Paleogene fossils has helped to date the major evolutionary radiations. We collate this information and then add data from the natural history of the modern fauna to sketch a history of major ecological adaptations at the subfamily level. The key events appear to have been, first, a mid-Cretaceous initial radiation in forest ground litter and soil coincident with the rise of the angiosperms (flowering plants), then a Paleogene advance to ecological dominance in concert with that of the angiosperms in tropical forests, and, finally, an expansion of some of the lineages, aided by changes in diet away from dependence on predation, upward into the canopy, and outward into more xeric environments. PMID- 15899977 TI - Demasking biological oscillators: properties and principles of entrainment exemplified by the Neurospora circadian clock. AB - Oscillations are found throughout the physical and biological worlds. Their interactions can result in a systematic process of synchronization called entrainment, which is distinct from a simple stimulus-response pattern. Oscillators respond to stimuli at some times in their cycle and may not respond at others. Oscillators can also be driven if the stimulus is strong (or if the oscillator is weak); i.e., they restart their cycle every time they receive a stimulus. Stimuli can also directly affect rhythms without entraining the underlying oscillator (masking): Drivenness and masking are often difficult to distinguish. Here we use the circadian biological clock to explore properties of entrainment. We confirm previous results showing that the residual circadian system in Neurospora can be entrained in a mutant of the clock gene frequency (frq(9), a strain deficient in producing a functional FRQ protein). This finding has implications for understanding the evolution of circadian programs. By comparing data sets from independent studies, we develop a template for analyzing, modeling, and dissecting the interactions of entrained and masked components. These insights can be applied to oscillators of all periodicities. PMID- 15899978 TI - DksA potentiates direct activation of amino acid promoters by ppGpp. AB - Amino acid starvation in Escherichia coli results in a spectrum of changes in gene expression, including inhibition of rRNA and tRNA promoters and activation of certain promoters for amino acid biosynthesis and transport. The unusual nucleotide ppGpp plays an important role in both negative and positive regulation. Previously, we and others suggested that positive effects of ppGpp might be indirect, resulting from the inhibition of rRNA transcription and, thus, liberation of RNA polymerase for binding to other promoters. Recently, we showed that DksA binds to RNA polymerase and greatly enhances direct effects of ppGpp on the negative control of rRNA promoters. This conclusion prompted us to reevaluate whether ppGpp might also have a direct role in positive control. We show here that ppGpp greatly increases the rate of transcription initiation from amino acid promoters in a purified system but only when DksA is present. Activation occurs by stimulation of the rate of an isomerization step on the pathway to open complex formation. Consistent with the model that ppGpp/DksA stimulates amino acid promoters both directly and indirectly in vivo, cells lacking dksA fail to activate transcription from the hisG promoter after amino acid starvation. Our results illustrate how transcription factors can positively regulate transcription initiation without binding DNA, demonstrate that dksA directly affects promoters in addition to those for rRNA, and suggest that some of the pleiotropic effects previously associated with dksA might be ascribable to direct effects of dksA on promoters involved in a wide variety of cellular functions. PMID- 15899979 TI - A vital role of tubulin-tyrosine-ligase for neuronal organization. AB - Tubulin is subject to a special cycle of detyrosination/tyrosination in which the C-terminal tyrosine of alpha-tubulin is cyclically removed by a carboxypeptidase and readded by a tubulin-tyrosine-ligase (TTL). This tyrosination cycle is conserved in evolution, yet its physiological importance is unknown. Here, we find that TTL suppression in mice causes perinatal death. A minor pool of tyrosinated (Tyr-)tubulin persists in TTL null tissues, being present mainly in dividing TTL null cells where it originates from tubulin synthesis, but it is lacking in postmitotic TTL null cells such as neurons, which is apparently deleterious because early death in TTL null mice is, at least in part, accounted for by a disorganization of neuronal networks, including a disruption of the cortico-thalamic loop. Correlatively, cultured TTL null neurons display morphogenetic anomalies including an accelerated and erratic time course of neurite outgrowth and a premature axonal differentiation. These anomalies may involve a mislocalization of CLIP170, which we find lacking in neurite extensions and growth cones of TTL null neurons. Our results demonstrate a vital role of TTL for neuronal organization and suggest a requirement of Tyr-tubulin for proper control of neurite extensions. PMID- 15899980 TI - Dissociation of an antiviral compound from the internal pocket of human rhinovirus 14 capsid. AB - WIN antiviral compounds bind human rhinovirus, as well as enterovirus and parechovirus, in an internal cavity located within the viral protein capsid. Access to the buried pocket necessitates deviation from the average viral protein structure identified by crystallography. We investigated the dissociation of WIN 52084 from the pocket in human rhinovirus 14 by using an adiabatic, biased molecular dynamics simulation method. Multiple dissociation trajectories are used to characterize the pathway. WIN 52084 exits between the polypeptide chain near the ends of betaC and betaH in a series of steps. Small, transient packing defects in the protein are sufficient for dissociation. A number of torsion-angle transitions of the antiviral compound are involved, which suggests that flexibility in antiviral compounds is important for binding. It is interesting to note that dissociation is associated with an increase in the conformational fluctuations of residues never in direct contact with WIN 52084 over the course of dissociation. These residues are N-terminal residues in the viral proteins VP3 and VP4 and are located in the interior of the capsid near the icosahedral 5-fold axis. The observed changes in dynamics may be relevant to structural changes associated with virion uncoating and its inhibition by antiviral compounds. PMID- 15899981 TI - Sca-1 expression identifies stem cells in the proximal region of prostatic ducts with high capacity to reconstitute prostatic tissue. AB - We previously showed that prostatic stem cells are concentrated in the proximal regions of prostatic ducts. We now report that these stem cells can be purified from isolated proximal duct regions by virtue of their high expression of the cell surface protein stem cell antigen 1 (Sca-1). In an in vivo prostate reconstitution assay, the purified Sca-1-expressing cell population isolated from the proximal region of ducts was more effective in generating prostatic tissue than a comparable population of Sca-1-depleted cells (203.0 +/- 83.1 mg vs. 11.9 +/- 9.2 mg) or a population of Sca-1-expressing cells isolated from the remaining regions of ducts (transit-amplifying cells) (31.9 +/- 24.1 mg). Almost all of the proliferative capacity of the proximal duct Sca-1-expressing cell population resides within the fraction of cells that express high levels of Sca-1 (top one third), with the proximal region of prostatic ducts containing 7.2-fold more Sca 1(high) cells than the remaining regions. More than 60% of the high-expressing cells coexpress alpha6 integrin and the anti-apoptotic factor Bcl-2, markers that are also characteristic of stem cells of other origins. Further stratification of the phenotype of the stem cells may enable the development of rational therapies for treating prostate cancer and benign prostatic hyperplasia. PMID- 15899982 TI - Cooperation between platelet-derived CD154 and CD4+ T cells for enhanced germinal center formation. AB - It has been demonstrated previously that platelet-derived CD154 communicates with the adaptive immune compartment, enhancing B and T cell responses in CD154(-/-) mice. The presence of platelets was also shown to be necessary for optimal production of immunoglobulin G (IgG) in normal C57BL/6 mice. These data led us to hypothesize that platelets perform a sentinel function, quickly relaying activating signals to the adaptive immune compartment. Here, we report that platelet-derived CD154 increases serum IgG levels and germinal center formation under conditions where antigen-specific CD4(+) T cell numbers are limiting. We propose that in the physiologic setting where antigen-specific B and T cells are rare, platelets function to enhance signals required for robust adaptive humoral immunity. PMID- 15899983 TI - Signaling through CD16b in human neutrophils involves the Tec family of tyrosine kinases. AB - Tec kinases belong to the second largest family of nonreceptor tyrosine kinases. Although these kinases are expressed in myeloid cells, little is known about their implication in neutrophil function. We recently reported the participation of Tec kinases in the responses of human neutrophils to the bacterial peptide N formyl-l-methionyl-l-leucyl-l-phenylalanine via G-coupled protein receptors. In this study, we extended our investigations of Tec kinases to the signaling of the glycosylphosphatidylinositol-linked receptor CD16b, which is highly and specifically expressed in neutrophils. The results obtained indicate that Tec is translocated to the plasma membrane, phosphorylated, and activated upon CD16b cross-linking and that the activation of Tec is inhibited by Src-specific inhibitors as well as by the phosphatidylinositol-3 kinase inhibitor, wortmannin. As no specific inhibitor of Tec exists, the role of Tec kinases was further investigated using a-Cyano-b-hydroxy-b-methyl-N-(2,5-dibromophenyl)propenamide (LFM-A13), a compound known to inhibit Bruton's tyrosine kinase. We show that this compound also inhibits the kinase activity of Tec and provide evidence that the mobilization of intracellular calcium and the tyrosine phosphorylation of phospholipase Cgamma2 (PLCgamma2) induced upon CD16b engagement are inhibited by LFM-A13. We also show that Tec kinases are important for CD16b-dependent degranulation of neutrophils. In summary, we provide direct evidence for the implication of Tec in CD16b signaling and suggest that Tec kinases are involved in the phosphorylation and activation of PLCgamma2 and subsequently, in the mobilization of calcium in human neutrophils. PMID- 15899984 TI - Differential and additive effects of platelet-derived chemokines on monocyte arrest on inflamed endothelium under flow conditions. AB - Platelet-derived chemokines, such as regulated on activation, normal T expressed and secreted (RANTES; CC chemokine ligand 5), platelet factor 4 [PF4; CXC chemokine ligand 4 (CXCL4)], and epithelial neutrophil-activating protein 78 (ENA 78; CXCL5), or precursors, such as beta-thromboglobulin, which can be processed to neutrophil-activating protein-2 (NAP-2; CXCL7), may play an important role in monocyte recruitment during atherogenesis. Platelets can deposit chemokines on inflamed endothelium; however, little is known about differential or additive effects of platelet chemokines on monocyte arrest. Here, we demonstrate that preincubation of activated human microvascular endothelial cells (HMVECs) with RANTES, PF4, or NAP-2 but not ENA-78 dose-dependently increased surface immobilization and subsequent monocyte arrest in flow. RANTES was the most potent and efficient arrest chemokine. Pretreatment of HMVECs with beta-thromboglobulin enhanced monocyte arrest in the presence of cathepsin G generating NAP-2. Combined pretreatment of HMVECs with RANTES and PF4 at suboptimal concentrations synergistically increased arrest, and preincubation with chondroitinase ABC abrogated RANTES- and PF4-induced monocyte arrest. This was associated with reduced expression of chondroitin sulfate, RANTES, and PF4 on the HMVEC surface. Perfusion of HMVECs with platelets known to deposit RANTES and PF4 on the endothelial surface enhanced monocyte arrest, which was inhibited by Met-RANTES, chondroitinase, or a blocking antibody to PF4 but not to ENA-78. The relevance of platelet-derived chemokines was confirmed in adhesion assays with activated whole blood, where Met-RANTES and to a lesser extent, antibodies to PF4 and NAP-2 inhibited arrest of CD14-positive monocytes. Thus, multiple platelet-derived chemokines and processable precursors, which can be presented by specific endothelial proteoglycans, may contribute and cooperate differentially to induce monocyte recruitment. PMID- 15899985 TI - The cover. Four Jockeys. PMID- 15899986 TI - A piece of my mind. Saying good-bye. PMID- 15899987 TI - Pediatric sleep medicine comes of age. PMID- 15899988 TI - NIH panel tries to clear confusion, spur research on managing menopause. PMID- 15899989 TI - Systemic tools can improve cardiac care. PMID- 15899990 TI - Cord blood stem cell network proposed. PMID- 15899997 TI - Holes in the Swiss health care system. PMID- 15899998 TI - Holes in the Swiss health care system. PMID- 15899999 TI - Holes in the Swiss health care system. PMID- 15900000 TI - Holes in the Swiss health care system. PMID- 15900001 TI - Holes in the Swiss health care system. PMID- 15900002 TI - Holes in the Swiss health care system. PMID- 15900003 TI - Tobacco brand appearances in movies before and after the master settlement agreement. PMID- 15900004 TI - Risk of celiac disease autoimmunity and timing of gluten introduction in the diet of infants at increased risk of disease. AB - CONTEXT: While gluten ingestion is responsible for the signs and symptoms of celiac disease, it is not known what factors are associated with initial appearance of the disease. OBJECTIVE: To examine whether the timing of gluten exposure in the infant diet was associated with the development of celiac disease autoimmunity (CDA). DESIGN, SETTING, AND PATIENTS: Prospective observational study conducted in Denver, Colo, from 1994-2004 of 1560 children at increased risk for celiac disease or type 1 diabetes, as defined by possession of either HLA-DR3 or DR4 alleles, or having a first-degree relative with type 1 diabetes. The mean follow-up was 4.8 years. MAIN OUTCOME MEASURE: Risk of CDA defined as being positive for tissue transglutaminase (tTG) autoantibody on 2 or more consecutive visits or being positive for tTG once and having a positive small bowel biopsy for celiac disease, by timing of introduction of gluten-containing foods into the diet. RESULTS: Fifty-one children developed CDA. Findings adjusted for HLA-DR3 status indicated that children exposed to foods containing wheat, barley, or rye (gluten-containing foods) in the first 3 months of life (3 [6%] CDA positive vs 40 [3%] CDA negative) had a 5-fold increased risk of CDA compared with children exposed to gluten-containing foods at 4 to 6 months (12 [23%] CDA positive vs 574 [38%] CDA negative) (hazard ratio [HR], 5.17; 95% confidence interval [CI], 1.44-18.57). Children not exposed to gluten until the seventh month or later (36 [71%] CDA positive vs 895 [59%] CDA negative) had a marginally increased risk of CDA compared with those exposed at 4 to 6 months (HR, 1.87; 95% CI, 0.97-3.60). After restricting our case group to only the 25 CDA-positive children who had biopsy-diagnosed celiac disease, initial exposure to wheat, barley, or rye in the first 3 months (3 [12%] CDA positive vs 40 [3%] CDA negative) or in the seventh month or later (19 [76%] CDA positive vs 912 [59%] CDA negative) significantly increased risk of CDA compared with exposure at 4 to 6 months (3 [12%] CDA positive vs 583 [38%] CDA negative) (HR, 22.97; 95% CI, 4.55-115.93; P = .001; and HR, 3.98; 95% CI, 1.18-13.46; P = .04, respectively). CONCLUSION: Timing of introduction of gluten into the infant diet is associated with the appearance of CDA in children at increased risk for the disease. PMID- 15900005 TI - Thrombophilia, clinical factors, and recurrent venous thrombotic events. AB - CONTEXT: Data on the recurrence rate of venous thrombotic events and the effect of several risk factors, including thrombophilia, remain controversial. The potential benefit of screening for thrombophilia with respect to prophylactic strategies and duration of anticoagulant treatment is not yet known. OBJECTIVES: To estimate the recurrence rate of thrombotic events in patients after a first thrombotic event and its determinants, including thrombophilic abnormalities. DESIGN, SETTING, AND PATIENTS: Prospective follow-up study of 474 consecutive patients aged 18 to 70 years without a known malignancy treated for a first objectively confirmed thrombotic event at anticoagulation clinics in the Netherlands. The Leiden Thrombophilia Study (LETS) was conducted from 1988 through 1992 and patients were followed up through 2000. MAIN OUTCOME MEASURES: Recurrent thrombotic event based on thrombophilic risk factors, sex, type of initial thrombotic event (idiopathic or provoked), oral contraceptive use, elevated levels of factors VIII, IX, XI, fibrinogen, homocysteine, and anticoagulant deficiencies. RESULTS: A total of 474 patients were followed up for mean (SD) of 7.3 (2.7) years and complete follow-up was achieved in 447 (94%). Recurrence of thrombotic events occurred in 90 patients during a total of 3477 patient-years. The rate of thrombotic event recurrence was 25.9 per 1000 patient years (95% confidence interval [CI], 20.8-31.8 per 1000 patient-years). The incidence rate of recurrence was highest during the first 2 years (31.9 per 1000 patient-years; 95% CI, 20.3-43.5 per 1000 patient-years). The risk of thrombotic event recurrence was 2.7 times (95% CI, 1.8-4.2 times) higher in men than in women. Patients whose initial thrombotic event was idiopathic had a higher risk of a thrombotic event recurrence than patients whose initial event was provoked (hazard ratio [HR], 1.9; 95% CI, 1.2-2.9). Women who used oral contraceptives during follow-up had a higher thrombotic event recurrence rate (28.0 per 1000 patient-years; 95% CI, 15.9-49.4 per 1000 patient-years) than those who did not (12.9 per 1000 patient-years; 95% CI, 7.9-21.2 per 1000 patient-years). Recurrence risks of a thrombotic event by laboratory abnormality ranged from an HR of 0.6 (95% CI, 0.3-1.1) in patients with elevated levels of factor XI to an HR of 1.8 (95% CI, 0.9-3.7) for patients with anticoagulant deficiencies. CONCLUSIONS: Prothrombotic abnormalities do not appear to play an important role in the risk of a recurrent thrombotic event. Testing for prothrombotic defects has little consequence with respect to prophylactic strategies. Clinical factors are probably more important than laboratory abnormalities in determining the duration of anticoagulation therapy. PMID- 15900006 TI - Relative citation impact of various study designs in the health sciences. AB - CONTEXT: The relative merits of various study designs and their placement in hierarchies of evidence are often discussed. However, there is limited knowledge about the relative citation impact of articles using various study designs. OBJECTIVE: To determine whether the type of study design affects the rate of citation in subsequent articles. DESIGN AND SETTING: We measured the citation impact of articles using various study designs--including meta-analyses, randomized controlled trials, cohort studies, case-control studies, case reports, nonsystematic reviews, and decision analysis or cost-effectiveness analysis- published in 1991 and in 2001 for a sample of 2646 articles. MAIN OUTCOME MEASURE: The citation count through the end of the second year after the year of publication and the total received citations. RESULTS: Meta-analyses received more citations than any other study design both in 1991 (P<.05 for all comparisons) and in 2001 (P<.001 for all comparisons) and both in the first 2 years and in the longer term. More than 10 citations in the first 2 years were received by 32.4% of meta-analyses published in 1991 and 43.6% of meta-analyses published in 2001. Randomized controlled trials did not differ significantly from epidemiological studies and nonsystematic review articles in 1991 but clearly became the second-cited study design in 2001. Epidemiological studies, nonsystematic review articles, and decision and cost-effectiveness analyses had relatively similar impact; case reports received negligible citations. Meta analyses were cited significantly more often than all other designs after adjusting for year of publication, high journal impact factor, and country of origin. When limited to studies addressing treatment effects, meta-analyses received more citations than randomized trials. CONCLUSION: Overall, the citation impact of various study designs is commensurate with most proposed hierarchies of evidence. PMID- 15900007 TI - Life-threatening sepsis associated with adjuvant doxorubicin plus docetaxel for intermediate-risk breast cancer. AB - CONTEXT: Adjuvant chemotherapy with new cytotoxic agents for breast cancer must be properly assessed for toxicity. OBJECTIVE: To describe adverse events associated with adjuvant chemotherapy for breast cancer, which led to premature termination of a clinical trial. DESIGN, SETTING, AND PATIENTS: We conducted a prospective randomized multicenter study (Reposant sur des Arguments Pronostiques et Predictifs [RAPP]-01) to compare the effectiveness of 2 chemotherapy regimens. Patients (women aged 18-70 years) had primary unilateral breast cancer and either a moderate number of positive axillary lymph nodes (< or =3) or no positive axillary lymph nodes (N0), but were at a high risk of relapse. Patients were treated at 11 French cancer referral centers from June 1999 through January 2003. Primary prophylaxis for febrile neutropenia was not recommended in the study protocol. INTERVENTIONS: Doxorubicin, 50 mg/m2, plus docetaxel, 75 mg/m2, or doxorubicin, 60 mg/m2, plus cyclophosphamide, 600 mg/m2, given postoperatively for 4 courses. MAIN OUTCOME MEASURES: The main end point was the disease-free survival rate at 5 years, as estimated using the Kaplan-Meier product limit method. Secondary end points included safety, which is the focus of this article, and overall survival. RESULTS: A total of 627 women were enrolled. Median follow up is currently too short (24 months) to analyze the primary end point. The trial was terminated prematurely when 2 deaths related to drug toxicity and 1 case of perforative peritonitis occurred among patients with febrile neutropenia, all in the doxorubicin-docetaxel group. The incidence of febrile neutropenia was significantly higher with the doxorubicin-docetaxel regimen (40.8%) than with the doxorubicin-cyclophosphamide regimen (7.1%) (P<.001). CONCLUSIONS: A high risk of life-threatening complications associated with the doxorubicin-docetaxel regimen was found in this open-label controlled trial. The doxorubicin-docetaxel combination should not be considered as an alternative to the doxorubicin cyclophosphamide regimen outside carefully designed studies that include primary prophylaxis for febrile neutropenia. PMID- 15900008 TI - Neonatal signs after late in utero exposure to serotonin reuptake inhibitors: literature review and implications for clinical applications. AB - CONTEXT: A neonatal behavioral syndrome linked to in utero serotonin reuptake inhibitor (SRI) exposure during the last trimester of pregnancy has been identified. The US Food and Drug Administration (FDA) and drug manufacturers have recently agreed to a class labeling change for SRIs, which include selective serotonin reuptake inhibitors (SSRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs), to include information about potential adverse events in neonates exposed in utero. Integration of data about the neonatal behavioral syndrome into the management of pregnancy in women who take SRIs is a current challenge for physicians. OBJECTIVES: To review evidence regarding the SRI related neonatal syndrome and to help clinicians guide their patients in a risk benefit decision-making process. DATA SOURCES: We searched MEDLINE (1966-February 2005) and PsycINFO (1974-February 2005). All articles related to neonatal signs after in utero SRI exposure were acquired, as well as unpublished data on this topic from the FDA advisory committee meeting of June 2004. References cited in case reports and studies were reviewed. Foreign-language literature was included and translated to English. STUDY SELECTION AND DATA EXTRACTION: Studies were included if they had clearly identified maternal SRI exposure for a minimum of the final trimester of pregnancy through delivery and assessed neonatal outcomes. We identified 13 case reports describing a total of 18 cases. Nine cohort studies met criteria. When not included in the published article, relative risks and 95% confidence intervals (CIs) were computed from raw data and summary risk ratios and 95% CIs were determined with Mantel-Haenszel estimates. DATA SYNTHESIS: Compared with early gestational SRI exposure or no exposure, late SRI exposure carries an overall risk ratio of 3.0 (95% CI, 2.0-4.4) for a neonatal behavioral syndrome. The most SRI-related neonatal case reports involved fluoxetine and paroxetine exposures. Neonates primarily display central nervous system, motor, respiratory, and gastrointestinal signs that are usually mild and disappear by 2 weeks of age. Medical management has consisted primarily of supportive care in special care nurseries. A severe syndrome that consists of seizures, dehydration, excessive weight loss, hyperpyrexia, or intubation is rare in term infants (1/313 quantifiable cases). There have been no reported neonatal deaths attributable to neonatal SRI exposure. CONCLUSIONS: Available evidence indicates that in utero exposure to SRIs during the last trimester through delivery may result in a self limited neonatal behavioral syndrome that can be managed with supportive care. The risks and benefits of discontinuing an SRI during pregnancy need to be carefully weighed for each individual patient. Development and validation of assessment methods and clinical management strategies are critical to advancing this research. PMID- 15900009 TI - Five years after To Err Is Human: what have we learned? AB - Five years ago, the Institute of Medicine (IOM) called for a national effort to make health care safe. Although progress since then has been slow, the IOM report truly "changed the conversation" to a focus on changing systems, stimulated a broad array of stakeholders to engage in patient safety, and motivated hospitals to adopt new safe practices. The pace of change is likely to accelerate, particularly in implementation of electronic health records, diffusion of safe practices, team training, and full disclosure to patients following injury. If directed toward hospitals that actually achieve high levels of safety, pay for performance could provide additional incentives. But improvement of the magnitude envisioned by the IOM requires a national commitment to strict, ambitious, quantitative, and well-tracked national goals. The Agency for Healthcare Research and Quality should bring together all stakeholders, including payers, to agree on a set of explicit and ambitious goals for patient safety to be reached by 2010. PMID- 15900010 TI - Is this patient having a stroke? AB - CONTEXT: Patients suspected of having a stroke or transient ischemic attack require accurate assessment for appropriate acute treatment and use of secondary preventive interventions. OBJECTIVE: To update a 1994 systematic review of the accuracy and reliability of symptoms and findings on neurological examination for the evaluation of patients with suspected stroke or transient ischemic attack. DATA SOURCES: We identified potential articles dated between 1994 and 2005 by multiple search strategies of the MEDLINE database and review of article and textbook bibliographies along with private collections. STUDY SELECTION: Selected articles provided primary data or appropriate summary statistics of the accuracy and/or reliability of the history or physical examination for diagnosis or short term prognosis of patients with neurological signs prompting a consideration of stroke. Articles addressing accuracy also needed to provide a final diagnosis following neuroimaging and all relevant laboratory tests. DATA EXTRACTION: The authors reviewed and abstracted data for estimating sensitivities, specificities, positive and negative likelihood ratios (LRs). Reliability assessment was based on reported kappa (kappa) statistics or intraclass correlation coefficients as appropriate. DATA SYNTHESIS: The prior probability of a stroke among patients with neurologically relevant symptoms is 10%. Based on studies using modern neuroimaging, the presence of acute facial paresis, arm drift, or abnormal speech increases the likelihood of stroke (LR of > or =1 finding = 5.5; 95% CI, 3.3 9.1), while the absence of all 3 decreases the odds (LR of 0 findings = 0.39; 95% CI, 0.25-0.61). The accurate determination of stroke subtype requires neuroimaging to distinguish ischemic from hemorrhagic stroke. Early mortality increases among those with any combination of impaired consciousness, hemiplegia, and conjugate gaze palsy (LR of > or =1 finding = 1.8; 95% CI, 1.2-2.8 and LR of 0 findings = 0.36; 95% CI, 0.13-1.0). Symptoms associated with high agreement for the diagnosis of stroke or transient ischemic attack vs no vascular event are a sudden change in speech, visual loss, diplopia, numbness or tingling, paralysis or weakness, and non-orthostatic dizziness (kappa = 0.60; 95% CI, 0.52-0.68). The reliabilities of individual neurological findings vary from slight to almost perfect, but can be improved with standardized scoring systems such as the National Institutes of Health Stroke Scale. Based on examination findings, stroke vascular distribution can be determined with moderate to good reliability (kappa = 0.54; 95% CI, 0.39-0.68). CONCLUSIONS: The history and clinical findings provide the basis for evaluating patients with possible stroke and choosing appropriate treatments. Focusing on 3 findings (acute facial paresis, arm drift, or abnormal speech) might improve diagnostic accuracy and reliability. PMID- 15900011 TI - Ethics, the constitution, and the dying process: the case of Theresa Marie Schiavo. PMID- 15900012 TI - Perspectives on new recommendations for nonoccupational HIV postexposure prophylaxis. PMID- 15900013 TI - Infant gluten and celiac disease: too early, too late, too much, too many questions. PMID- 15900014 TI - JAMA patient page. Celiac disease. PMID- 15900015 TI - Characterization of novel CYP2A6 polymorphic alleles (CYP2A6*18 and CYP2A6*19) that affect enzymatic activity. AB - Genetic polymorphisms of CYP2A6 gene are known as a causal factor of the interindividual differences in nicotine metabolism. We found three novel CYP2A6 alleles. The CYP2A6(*)18A allele has a single nucleotide polymorphism (SNP) of A5668T (A1175T, Y392F) in exon 8. The CYP2A6(*)18B allele has synonymous SNPs of G51A (G51A), T5684C (T1191C), and T5702C (T1209C) in addition to A5668T (A1175T, Y392F). The CYP2A6(*)19 allele has the SNPs of A5668T (A1175T, Y392F), T6354C (intron 8), and T6558C (T1412C, I471T) as well as the conversion with the CYP2A7 sequence in the 3'-untranslated region, in which the latter two changes correspond to CYP2A6(*)7. Ethnic differences in the frequencies of these alleles were observed between whites, African-Americans, Japanese, and Koreans. Wild or variant CYP2A6 (CYP2A6(*)18, CYP2A6(*)19, and CYP2A6(*)7) were expressed in Escherichia coli. For coumarin 7-hydroxylation and 5-fluorouracil formation from tegafur, the K(m) values were increased, and V(max) values were decreased in CYP2A6.18 compared with those in CYP2A6.1, resulting in decreased clearance to 50 and 35% of that of the wild type, respectively. The K(m) and V(max) values for nicotine C-oxidation were both increased, resulting in no change of clearance. In CYP2A6.19, the effects on the coumarin 7-hydroxylation and 5-fluorouracil formation (increased K(m) and decreased V(max)) were prominent, resulting in decreased clearance to 8% of those of the wild type. For nicotine C-oxidation, the K(m) and V(max) values were both decreased, resulting in decreased clearance to 30% of that of the wild type. The changes of the kinetics in CYP2A6.19 were similar to those in CYP2A6.7. In vivo nicotine metabolism was evaluated in whites (n = 56) and Koreans (n = 40). Although the CYP2A6(*)18 and CYP2A6(*)19 alleles were found only heterozygously, a subject with CYP2A6(*)7/CYP2A6(*)19 showed a lower cotinine/nicotine ratio of the plasma concentration compared with homozygotes of the CYP2A6(*)1A, supporting the in vitro results that the CYP2A6(*)19 allele leads to decreased enzymatic activity. PMID- 15900016 TI - Molecular mechanisms regulating human CYP4B1 lung-selective expression. AB - Lung-selective cytochrome P450 expression is well recognized; however, little is known regarding regulatory mechanisms. To address this knowledge gap, transient expression of CYP4B1/luciferase constructs was used to identify a proximal, positively acting regulatory element, position -139 to -45, that functioned in all cells examined; a negatively acting element, position -457 to -216, that only functioned in HepG2 hepatoblastoma cells; and a distal, positively acting element, position -1087 to -1008, that functioned in A549 or BEAS-2B lung-derived cells but not HepG2 cells or 293 kidney-derived cells. Competitive electrophoretic mobility shift assays further localized specific A549, but not HepG2, nuclear protein binding to two sites within the distal element, CYP4B1 position -1052 to -1042 and -1026 to -1008. Several potential lung-selective transcription factor recognition sequences were identified within these elements. However, attempts to identify specific factor(s) were unsuccessful. In contrast, in vitro DNA/protein binding assays combined with transient expression and mutagenesis studies identified two functional Sephadex protein/Kruppel-like factor families of transcription factor sites within the proximal element (position -118 to -114 and position -77 to -73) that bound both Sephadex protein 1 (Sp1) and Sephadex protein 3 (Sp3) in vitro. Furthermore, Sp1-dependent synergistic regulation was observed in A549 cells involving the proximal and distal regulatory elements. Chromatin immunoprecipitation assays demonstrated binding of neither Sp1 nor Sp3 to the CYP4B1 proximal element in human liver tissue, whereas selective Sp1 binding was observed in human lung tissue. Thus, the composite findings are consistent with both the proximal Sp1 elements and the distal regulatory element acting to synergistically control CYP4B1 lung-selective expression. PMID- 15900017 TI - Functional involvement of rat organic anion transporter 2 (Slc22a7) in the hepatic uptake of the nonsteroidal anti-inflammatory drug ketoprofen. AB - Rat organic anion transporter 2 (rOat2, Slc22a7) is a sinusoidal multispecific organic anion transporter in the liver. The role of rOat2 in the hepatic uptake of drugs has not been thoroughly investigated yet. rOat2 substrates include nonsteroidal anti-inflammatory drugs, such as ketoprofen, indomethacin, and salicylate. In the present study, the uptake of ketoprofen, indomethacin, and salicylate by freshly isolated rat hepatocytes was characterized. The uptake of ketoprofen, indomethacin, and salicylate by hepatocytes was sodium-independent, and the rank order of their uptake activities was indomethacin > ketoprofen > salicylate. Kinetic analysis based on Akaike's Information Criterion suggested that the uptake of ketoprofen and indomethacin by hepatocytes consists of two saturable components and one nonsaturable one. The K(m) and V(max) values for the high- and low-affinity components for ketoprofen uptake were 0.84 and 97 microM and 35 and 1800 pmol/min/mg protein, respectively, whereas those for indomethacin were 1.1 and 140 microM and 130 and 16,000 pmol/min/mg protein, respectively. The K(m) values of the high-affinity component were similar to those for rOat2 (3.3 and 0.37 microM for ketoprofen and indomethacin, respectively). The uptake of ketoprofen by hepatocytes was significantly inhibited by probenecid and rOat2 inhibitors (indocyanine green, indomethacin, glibenclamide, and salicylate). Other inhibitors of rOatps (taurocholate and pravastatin) and rOat3 (pravastatin and p-aminohippurate) had a slight effect, but digoxin had no effect. These results suggest that rOat2 accounts partly for the hepatic uptake of ketoprofen and, presumably, indomethacin as a high-affinity site and that other transporters, such as rOatps, but not rOatp2, and rOat3, are also involved. PMID- 15900018 TI - The coordination of prostaglandin E2 production by sphingosine-1-phosphate and ceramide-1-phosphate. AB - The ability of pro-inflammatory cytokines such as interleukin-1beta (IL-1beta) to induce the major inflammatory mediator prostaglandin (PG) E(2) depends on the activation of two rate-limiting enzymes, phospholipase A(2) (PLA(2)) and cyclooxygenase 2 (COX-2). PLA(2) acts to generate arachidonic acid, which serves as the precursor substrate for COX-2 in the metabolic pathway leading to PGE(2) production. However, less is known about the mechanisms that coordinate the regulation of these two enzymes. We have provided prior evidence that sphingosine kinase 1 and its bioactive lipid product sphingosine-1-phosphate (S1P) mediate the effects of cytokines on COX-2 induction, whereas ceramide kinase and its distinct product, ceramide-1-phosphate (C1P), are required for the activation and translocation of cPLA(2) (FASEB J 17:1411-1421. 2003; J Biol Chem 278:38206 38213, 2003; J Biol Chem 279:11320-11326, 2004). Herein, we show that these two pathways are independent but coordinated, resulting in synergistic induction of PGE(2). Moreover, the combination of both S1P and C1P recapitulates the temporal and spatial activation of cPLA(2) and with COX-2 seen IL-1beta. Taken together, the results provide, for the first time, a mechanism that assures the coordinate expression and activation in time and space of COX-2 and cPLA(2), assuring maximal production of PGE(2). PMID- 15900019 TI - Activated extracellular signal-regulated kinases are necessary and sufficient to initiate tubulogenesis in renal tubular MDCK strain I cell cysts. AB - A classic in vitro model of renal cyst and tubule formation utilizes the Madin Darby canine kidney (MDCK) cell line, of which two strains exist. Most cyst and tubule formation studies that utilized MDCK cells have been performed with MDCK strain II cells. MDCK strain II cells form hollow cysts in a three-dimensional collagen matrix over 10 days and tubulate in response to hepatocyte growth factor, which increases levels of active (phosphorylated) ERK1/2. In this study, we demonstrate that MDCK strain I cells also form cysts when grown in a collagen matrix; however, MDCK strain I cell cysts spontaneously initiate the primary steps in tubulogenesis. Analysis of time-lapse microscopy of both MDCK strain I and strain II cell cysts during the initial stages of tubulogenesis demonstrates a highly dynamic process with cellular extensions and retractions occurring rapidly and continuously. MDCK strain I cell cysts can spontaneously initiate tubulogenesis mainly because of relatively higher levels of active ERK in MDCK strain I, compared with strain II, cells. The presence of either of two distinct inhibitors of ERK activation (UO126 and PD09059) prevents tubulogenesis from occurring spontaneously in MDCK strain I cell cysts and, in response to hepatocyte growth factor, in strain II cell cysts. The difference between MDCK strain I and strain II cell lines is likely explained by differing embryological origins, with strain I cells being of collecting duct, and hence ureteric bud, origin. Ureteric bud cells also have high levels of active ERK and spontaneously tubulate in our in vitro collagen gel system, with tubulogenesis inhibited by UO126 and PD09059. These results suggest that a seminal event in kidney development may be the activation of ERK in the mesonephric duct/ureteric bud cells destined to form the collecting tubules. PMID- 15900020 TI - Interactive effects of superoxide anion and nitric oxide on blood pressure and renal hemodynamics in transgenic rats with inducible malignant hypertension. AB - Superoxide anion contributes to the pathogenesis of various forms of hypertension, but its role in the development of malignant hypertension remains unclear. The present study was performed to determine the influence of superoxide anion on blood pressure and renal hemodynamics in transgenic rats with inducible malignant hypertension [strain name: TGR(Cyp1a1Ren2)]. Malignant hypertension was induced in male Cyp1a1-Ren2 rats (n = 6) through dietary administration of the aryl hydrocarbon, indole-3-carbinol (0.3%) for 7-9 days. Mean arterial pressure (MAP) and renal hemodynamics were measured in pentobarbital sodium-anesthetized Cyp1a1-Ren2 rats before and during intravenous infusion of the superoxide dismutase mimetic tempol (100 mumol/h). Basal MAP and renal vascular resistance (RVR) were elevated in rats induced with indole-3-carbinol compared with noninduced rats (n = 5) (184 +/- 4 vs. 127 +/- 3 mmHg, P < 0.01, and 29 +/- 2 vs. 21 +/- 1 mmHg.ml(-1).min.g, P < 0.01, respectively). Hypertensive rats had elevated excretion of urinary 8-isoprostane compared with normotensive rats (41 +/- 4 vs. 13 +/- 6 pg.min(-1).g(-1), P < 0.01). There were no differences in renal plasma flow and glomerular filtration rate between groups. Systemic administration of tempol decreased MAP (184 +/- 4 to 151 +/- 4 mmHg, P < 0.01) and RVR (29 +/- 2 to 25 +/- 2 mmHg.ml(-1).min.g, P < 0.05) in hypertensive but not in normotensive Cyp1a1-Ren2 rats. In addition, tempol administration decreased urinary excretion of 8-isoprostane (41 +/- 4 to 25 +/- 4 pg.min(-1).g( 1), P < 0.05). Renal plasma flow and glomerular filtration rate remained unaltered during tempol administration in both groups. The administration of the nitric oxide synthase inhibitor nitro-l-arginine attenuated the decrease in MAP and RVR in response to tempol. These findings indicate that superoxide anion contributes to the elevated RVR and increased arterial blood pressure, by a mechanism that is at least in part nitric oxide dependent, in Cyp1a1-Ren2 rats with malignant hypertension. PMID- 15900021 TI - Superiority of combination of thiazide with angiotensin-converting enzyme inhibitor or AT1-receptor blocker over thiazide alone on renoprotection in L NAME/SHR. AB - The renal and glomerular dynamic effects of combining thiazide and angiotensin antagonists have not been reported. The present study was designed to examine the effects of hydrochlorothiazide (HCTZ) alone or in combination with an angiotensin converting enzyme inhibitor or ANG II type 1-receptor blocker on renal hemodynamics, glomerular dynamics, renal function, and renal histopathology in the N(omega)-nitro-l-arginine methyl ester-treated spontaneously hypertensive rat (l-NAME/SHR) model. HCTZ (80 mg x kg(-1) x day(-1)) alone or in combination with enalapril (30 mg x kg(-1) x day(-1)) or losartan (30 mg x kg(-1) x day(-1)) or enalapril (15 mg.kg(-1).day(-1)) plus losartan (15 mg x kg(-1) x day(-1)) was administered to l-NAME/SHR (5.0 +/- 0.10 mg x kg(-1) x day(-1)) for 3 wk. Mean arterial pressure, total peripheral resistance, renal plasma flow, glomerular filtration rate, glomerular hydrostatic pressure, afferent and efferent glomerular arteriolar resistances, single nephron plasma flow, single nephron glomerular filtration rate, serum creatinine concentration, 24-h urinary protein excretion, and glomerular and arteriolar injury scores were determined. HCTZ reduced mean arterial pressure, total peripheral resistance, glomerular hydrostatic pressure, and afferent and efferent glomerular arteriolar resistances (P < 0.05, at least) but slightly increased renal plasma flow and single nephron plasma flow associated with reduced serum creatinine concentration, urinary protein excretion, and arteriolar injury score compared with l-NAME/SHR control. However, the combination of enalapril and/or losartan with HCTZ markedly improved each of these functions. These results demonstrated minor benefits of HCTZ monotherapy and a marked superiority of its combination with enalapril and/or losartan over HCTZ monotherapy on renoprotection in l-NAME/SHR, thereby providing strong evidence of their clinical benefits for hypertensive patients with renal functional impairment. PMID- 15900022 TI - Modeling GFR trajectories in diabetic nephropathy. AB - In an 8-year longitudinal study of Pima Indians with type 2 diabetes and nephropathy, we used statistical techniques that are novel and depend on minimal assumptions to compare longitudinal measurements of glomerular filtration rate (GFR). Individuals enrolled with new-onset microalbuminuria either progressed to macroalbuminuria (progressors, n = 13) or did not progress (nonprogressors, n = 13) during follow-up. Subjects with new-onset macroalbuminuria at screening were also followed (n = 22). Patients had their GFR determined serially by urinary iothalamate clearances (average 11 clearances; range 6-19). GFR courses of individuals were modeled using an adaptation of smoothing and regression cubic B splines. Group comparisons were based on five-component vectors of fitted GFR values using a permutation approach to a Hotelling's T(2) statistic. GFR profiles of initially microalbuminuric progressors differed significantly from those of nonprogressors (P = 0.003). There were no significant baseline differences between progressors and nonprogressors with respect to any measured clinical parameters. The course of GFR in the first 4 yr following progression to macroalbuminuria in initially microalbuminuric subjects did not differ from that in newly screened macroalbuinuric subjects (P = 0.27). Without imposing simplifying models on the data, the statistical techniques used demonstrate that the courses of decline of GFR in definable subgroups of initially microalbuminuric diabetic Pima Indians, although generally progressive, follow distinct trajectories that are related to the extent of glomerular barrier dysfunction, as reflected by the evolution from microalbuminuria to macroalbuminuria. PMID- 15900023 TI - SMP-534 inhibits TGF-beta-induced ECM production in fibroblast cells and reduces mesangial matrix accumulation in experimental glomerulonephritis. AB - Transforming growth factor-beta (TGF-beta) is a potent fibrotic factor responsible for the synthesis of extracellular matrix (ECM) and is implicated as the major determinant in pathogenesis of chronic fibroses, including kidney. The novel small compound SMP-534 reduced ECM production induced by TGF-beta in fibroblast cells. SMP-534 inhibited TGF-beta-induced p38 mitogen-activated protein kinase (p38) activation but did not inhibit epidermal growth factor (EGF) induced extracellular signal-related kinase (ERK) activation. We also found that oral administration of SMP-534 dose dependently lowered hydroxyproline contents in the cortical region of the kidney in rat anti-Thy-1 nephritis models. In periodic acid-Schiff staining of kidney sections, ECM accumulation was reduced by SMP-534 treatment. These data indicate that SMP-534 has potential in therapy for fibrotic diseases, including nephropathy. PMID- 15900024 TI - High NaCl increases TonEBP/OREBP mRNA and protein by stabilizing its mRNA. AB - Hypertonicity increases mRNA and protein abundance of the transcription factor tonicity-responsive enhancer/osmotic response element binding protein (TonEBP/OREBP), contributing to increased transcription of downstream osmoprotective genes. Previously, this was attributed to increased transcription of TonEBP/OREBP because no change was found in its mRNA stability. However, there is no direct evidence for increased transcription, and the 3'-untranslated region (UTR) of TonEBP/OREBP contains numerous adenylate/uridylate-rich elements, which can modulate RNA stability. Therefore, we have reinvestigated the effect of hypertonicity on TonEBP/OREBP mRNA stability. We find that, in mouse inner medullary collecting duct cells, raising osmolality from 300 to 500 mosmol/kgH(2)O by adding NaCl increases TonEBP/OREBP mRNA to a peak of 2.3-fold after 4 h, followed by a decline. TonEBP/OREBP protein increases to a sustained peak of 3.0-fold at 8 h. To determine the stability of TonEBP/OREBP mRNA, we measured the rate of its decrease after inhibiting transcription with actinomycin D, finding that it is stabilized for 6 h after addition of NaCl. This stabilization is sufficient to explain the increase in mRNA without any change in transcription. To investigate how hypertonicity stabilizes TonEBP/OREBP mRNA, we tested luciferase reporters containing parts of the TonEBP/OREBP mRNA UTR. Inclusion of both the 5'- and 3'-UTR increases reporter activity, consistent with mRNA stabilization. Surprisingly, however, it is the 5'-UTR that stabilizes; the 3'-UTR, by itself, decreases reporter activity. We concluded that 1) hypertonicity stabilizes TonEBP/OREBP mRNA, contributing to its increase, and 2) stabilization depends on the presence of the 5'-UTR. PMID- 15900025 TI - Dysfunction of the brain calcium channel CaV2.1 in absence epilepsy and episodic ataxia--a comment. PMID- 15900027 TI - Altered central excitability and analgesic treatment in patients with restless legs syndrome. PMID- 15900030 TI - Commentary on: Characteristics of two distinct clinical phenotypes in pathologically proven progressive supranuclear palsy: Richardson's syndrome and PSP-parkinsonism, by D. Williams, R. de Silva, D. Paviour, et al. (Brain-2004 01045.R1). PMID- 15900031 TI - A rationally designed small molecule that inhibits the HIF-1alpha-ARNT heterodimer from binding to DNA in vivo. AB - Modern drug development is focused on two steps: the identification of new molecular targets and the development of drugs that affect these targets. A molecular target can be an enzymatic activity or a macromolecular interface that is important in a disease pathway. Current drugs on the market are biased toward targeting cell surface receptors and intracellular enzymatic activities. However, macromolecular interfaces can also serve as potential molecular targets. A recent paper from Kaelin and Dervan's groups examined an underused molecular target transcription factor DNA binding. To specifically disrupt transcriptional activation, they used a rationally designed small molecule that binds specifically in the minor groove of a DNA sequence that in vivo is bound by a bHLH heterodimer transcription factor. PMID- 15900032 TI - Regulating inducible transcription through controlled localization. AB - Many signaling pathways regulate the activity of effector transcription factors by controlling their subcellular localization. Until recently, the cytoplasmic retention of inactive transcription factors was mainly attributed to binding partners that mask the nuclear localization signals (NLSs) of target proteins. Inactive transcription factors were thought to be exclusively cytoplasmic until their activation, after which the NLSs were unmasked to allow nuclear translocation. There is now a growing body of evidence, however, that challenges this simple model. This review discusses recent reports that suggest that inducible transcription factors can constantly shuttle between the cytoplasm and the nucleus, and that their apparent cytoplasmic retention can be achieved by binding partners that mask the NLSs, tether the transcription factor to cytoplasmic structures, or mark the transcription factor for proteasomal degradation. We also discuss the possibility that this more complex model of cytoplasmic retention might be applicable to a broader range of transcription factors and their associated signaling pathways. PMID- 15900034 TI - Radiation-induced thyroid cancer--what's new? PMID- 15900035 TI - Targeting coagulation to the tumor microvasculature: perspectives and therapeutic implications from preclinical studies. PMID- 15900036 TI - New office and new leader aim to streamline FDA cancer drug review process. PMID- 15900037 TI - NCCN guidelines advocate wider use of colony-stimulating factor. PMID- 15900038 TI - Cancer pain management guidelines issued for children; adult guidelines updated. PMID- 15900039 TI - Stat bite: Change in incidence of selected cancers in Great Britain, 1991-2000. PMID- 15900040 TI - Cancer research funding in Europe low compared with U.S., survey finds. PMID- 15900041 TI - Cancer risk prediction models: a workshop on development, evaluation, and application. AB - Cancer researchers, clinicians, and the public are increasingly interested in statistical models designed to predict the occurrence of cancer. As the number and sophistication of cancer risk prediction models have grown, so too has interest in ensuring that they are appropriately applied, correctly developed, and rigorously evaluated. On May 20-21, 2004, the National Cancer Institute sponsored a workshop in which experts identified strengths and limitations of cancer and genetic susceptibility prediction models that were currently in use and under development and explored methodologic issues related to their development, evaluation, and validation. Participants also identified research priorities and resources in the areas of 1) revising existing breast cancer risk assessment models and developing new models, 2) encouraging the development of new risk models, 3) obtaining data to develop more accurate risk models, 4) supporting validation mechanisms and resources, 5) strengthening model development efforts and encouraging coordination, and 6) promoting effective cancer risk communication and decision-making. PMID- 15900042 TI - Risk of thyroid cancer after exposure to 131I in childhood. AB - BACKGROUND: After the Chernobyl nuclear power plant accident in April 1986, a large increase in the incidence of childhood thyroid cancer was reported in contaminated areas. Most of the radiation exposure to the thyroid was from iodine isotopes, especially 131I. We carried out a population-based case-control study of thyroid cancer in Belarus and the Russian Federation to evaluate the risk of thyroid cancer after exposure to radioactive iodine in childhood and to investigate environmental and host factors that may modify this risk. METHODS: We studied 276 case patients with thyroid cancer through 1998 and 1300 matched control subjects, all aged younger than 15 years at the time of the accident. Individual doses were estimated for each subject based on their whereabouts and dietary habits at the time of the accident and in following days, weeks, and years; their likely stable iodine status at the time of the accident was also evaluated. Data were analyzed by conditional logistic regression using several different models. All statistical tests were two-sided. RESULTS: A strong dose response relationship was observed between radiation dose to the thyroid received in childhood and thyroid cancer risk (P<.001). For a dose of 1 Gy, the estimated odds ratio of thyroid cancer varied from 5.5 (95% confidence interval [CI] = 3.1 to 9.5) to 8.4 (95% CI = 4.1 to 17.3), depending on the risk model. A linear dose response relationship was observed up to 1.5-2 Gy. The risk of radiation-related thyroid cancer was three times higher in iodine-deficient areas (relative risk [RR]= 3.2, 95% CI = 1.9 to 5.5) than elsewhere. Administration of potassium iodide as a dietary supplement reduced this risk of radiation-related thyroid cancer by a factor of 3 (RR = 0.34, 95% CI = 0.1 to 0.9, for consumption of potassium iodide versus no consumption). CONCLUSION: Exposure to (131)I in childhood is associated with an increased risk of thyroid cancer. Both iodine deficiency and iodine supplementation appear to modify this risk. These results have important public health implications: stable iodine supplementation in iodine-deficient populations may substantially reduce the risk of thyroid cancer related to radioactive iodines in case of exposure to radioactive iodines in childhood that may occur after radiation accidents or during medical diagnostic and therapeutic procedures. PMID- 15900043 TI - Specific occlusion of murine and human tumor vasculature by VCAM-1-targeted recombinant fusion proteins. AB - BACKGROUND: The tumor vasculature is increasingly recognized as a target for cancer therapy. We developed and evaluated recombinant fusion proteins targeting the coagulation-inducing protein soluble tissue factor (sTF) to the luminal tumor endothelial antigen vascular cell adhesion molecule 1 (VCAM-1, CD106). METHODS: We generated fusion proteins consisting of sTF fused to antibody fragments directed against mouse or human VCAM-1 and characterized them in vitro by flow cytometry, surface plasmon resonance, and two-stage coagulation assays. Their therapeutic effects were tested in three human xenograft tumor models: L540rec Hodgkin lymphoma, Colo677 small-cell lung carcinoma, and Colo677/HDMEC small-cell lung carcinoma with human vasculature. Toxicity was analyzed by histologic examination of organs and determination of laboratory blood parameters. RESULTS: The fusion proteins bound VCAM-1 with nanomolar affinities and had the same coagulation activity as an sTF standard. Xenograft tumor-bearing mice treated with fusion protein (FP) alone or in combination with lipopolysaccharide (FP/L) or doxorubicin (FP/D) exhibited tumor-selective necrosis (L540rec tumors: 74% tumor necrosis [95% confidence interval {CI} = 55% to 93%] with FP/L versus 13% tumor necrosis [95% CI = 4% to 22%] with vehicle; Colo677 tumors: 26% [95% CI = 16% to 36%] with FP versus 8% [95% CI = 2% to 14%] with vehicle); tumor growth delay (Colo677/HDMEC: mean tumor weights after 3 days = 42 mg in FP-treated mice versus 71 mg in vehicle-treated mice, difference = 29 mg, 95% CI = 8 to 100, Mann Whitney P = .008); and some tumor regressions (one of seven FP-treated Colo677 tumor-bearing mice and two of seven FP/D-treated mice). The fusion protein was well tolerated. CONCLUSIONS: Recombinant tissue factor-based fusion proteins directed against an intraluminal tumor endothelial cell marker induce tumor selective intravascular coagulation, tumor tissue necrosis, and tumor growth delay. PMID- 15900044 TI - Effect of recall rate on earlier screen detection of breast cancers based on the Dutch performance indicators. AB - BACKGROUND: The recall rate (i.e., the rate at which mammographically screened women are recalled for additional assessment) in the Dutch breast screening program (0.89% in 2000 for subsequent examinations) is the lowest worldwide, with possible consequences including higher rates of late-detected (i.e., "missed") interval and screen-detected cancers. To estimate the effect of changes in recall rate on earlier detection of cancers, we carried out a blinded review of interval and screen-detected cancers in the Dutch screening program. METHODS: A total of 495 sets of screen-negative mammograms (prediagnostic mammogram and the immediate previous mammogram) were collected from women participating in the biennial Dutch screening program. Of these, 250 were from control subjects, and 245 were from women who were subsequently diagnosed with breast cancer (123 interval and 122 screen-detected cancers). These mammograms were read by 15 radiologists who specialize in screening mammography and were blinded to outcome. Mean detection sensitivities for different false-positive rates were calculated using a linear mixed model. These results were used to calculate the effect of recall rate adjustment on earlier detection of cancers and numbers of false-positives. RESULTS: Increasing the recall rate to 2.0% would increase the detection rate from 4.20 per thousand to 4.52 per thousand due to the earlier detection of interval cancers. Moreover, 0.54 per thousand of the screen-detected cancers would be detected 2 years earlier (late screen-detected cancers). At recall rates of 3.0% and 4.0% the detection rate would increase to 4.58 per thousand and 4.63 per thousand, respectively, and 0.64 per thousand and 0.72 per thousand, respectively, of the screen-detected cancers would be detected 2 years earlier. For each 1.0% incremental increase in recall rate above 5.0%, the detection rate would increase by approximately 0.03 per thousand, with positive predictive values decreasing to below 10%. CONCLUSION: Breast cancer can be detected earlier by lowering the threshold for recall, especially for recall rates of 1%-4%. With further recall rate increases, cancer detection levels off with a disproportionate increase of false-positive rates. PMID- 15900045 TI - Serum sex steroids in premenopausal women and breast cancer risk within the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - BACKGROUND: Contrasting etiologic hypotheses about the role of endogenous sex steroids in breast cancer development among premenopausal women implicate ovarian androgen excess and progesterone deficiency, estrogen excess, estrogen and progesterone excess, and both an excess or lack of adrenal androgens (dehydroepiandrosterone [DHEA] or its sulfate [DHEAS]) as risk factors. We conducted a case-control study nested within the European Prospective Investigation into Cancer and Nutrition cohort to examine associations among premenopausal serum concentrations of sex steroids and subsequent breast cancer risk. METHODS: Levels of DHEAS, (Delta4-)androstenedione, testosterone, and sex hormone binding globulin (SHBG) were measured in single prediagnostic serum samples from 370 premenopausal women who subsequently developed breast cancer (case patients) and from 726 matched cancer-free control subjects. Levels of progesterone, estrone, and estradiol were also measured for the 285 case patients and 555 matched control subjects who had provided information about the day of menstrual cycle at blood donation. Conditional logistic regression models were used to estimate relative risks of breast cancer by quartiles of hormone concentrations. All statistical tests were two-sided. RESULTS: Increased risks of breast cancer were associated with elevated serum concentrations of testosterone (odds ratio [OR] for highest versus lowest quartile = 1.73, 95% confidence interval [CI] = 1.16 to 2.57; P(trend) = .01), androstenedione (OR for highest versus lowest quartile = 1.56, 95% CI = 1.05 to 2.32; P(trend) = .01), and DHEAS (OR for highest versus lowest quartile = 1.48, 95% CI = 1.02 to 2.14; P(trend) = .10) but not SHBG. Elevated serum progesterone concentrations were associated with a statistically significant reduction in breast cancer risk (OR for highest versus lowest quartile = 0.61, 95% CI = 0.38 to 0.98; P(trend) = .06). The absolute risk of breast cancer for women younger than 40 followed up for 10 years was estimated at 2.6% for those in the highest quartile of serum testosterone versus 1.5% for those in the lowest quartile; for the highest and lowest quartiles of progesterone, these estimates were 1.7% and 2.6%, respectively. Breast cancer risk was not statistically significantly associated with serum levels of the other hormones. CONCLUSIONS: Our results support the hypothesis that elevated blood concentrations of androgens are associated with an increased risk of breast cancer in premenopausal women. PMID- 15900046 TI - p53-defective tumors with a functional apoptosome-mediated pathway: a new therapeutic target. AB - BACKGROUND: Although cancer cells appear to maintain the machinery for intrinsic apoptosis, defects in the pathway develop during malignant transformation, preventing apoptosis from occurring. How to specifically induce apoptosis in cancer cells remains unclear. METHODS: We determined the apoptosome activity and p53 status of normal human cells and of lung, colon, stomach, brain, and breast cancer cells by measuring cytochrome c-dependent caspase activation and by DNA sequencing, respectively, and we used COMPARE analysis to identify apoptosome specific agonists. We compared cell death, cytochrome c release, and caspase activation in NCI-H23 (lung cancer), HCT-15 (colon cancer), and SF268 (brain cancer) cells treated with Triacsin c, an inhibitor of acyl-CoA synthetase (ACS), or with vehicle. The cells were mock, transiently, or stably transfected with genes for Triacsin c-resistant ACSL5, dominant negative caspase-9, or apoptotic protease activating factor-1 knockdown. We measured ACS activity and levels of cardiolipin, a mitochondrial phospholipid, in mock and ACSL5-transduced SF268 cells. Nude mice carrying NCI-H23 xenograft tumors (n = 10) were treated with Triacsin c or vehicle, and xenograft tumor growth was assessed. Groups were compared using two-sided Student t tests. RESULTS: Of 21 p53-defective tumor cell lines analyzed, 17 had higher apoptosome activity than did normal cells. Triacsin c selectively induced apoptosome-mediated death in tumor cells (caspase activity of Triacsin c-treated versus untreated SF268 cells; means = 1020% and 100%, respectively; difference = 920%, 95% CI = 900% to 940%; P<.001). Expression of ACSL5 suppressed Triacsin c-induced cytochrome c release and subsequent cell death (cell survival of Triacsin c-treated mock- versus ACSL5-transduced SF268 cells; means = 40% and 83%, respectively; difference = 43%, 95% CI = 39% to 47%; P<.001). ACS was also essential to the maintenance of cardiolipin levels. Finally, Triacsin c suppressed growth of xenograft tumors (relative tumor volume on day 21 of Triacsin c-treated versus untreated mice; means = 4.6 and 9.6, respectively; difference = 5.0, 95% CI = 2.1 to 7.9; P = .006). CONCLUSIONS: Many p53-defective tumors retain activity of the apoptosome, which is therefore a potential target for cancer chemotherapy. Inhibition of ACS may be a novel strategy to induce the death of p53-defective tumor cells. PMID- 15900047 TI - Re: Integrin beta3 Leu33Pro homozygosity and risk of cancer. PMID- 15900048 TI - Re: Are breast density and bone mineral density independent risk factors for breast cancer? PMID- 15900050 TI - 20 years of patient dose studies: where should we go from here? PMID- 15900051 TI - Installation of a PACS system. PMID- 15900052 TI - Using PACS as a teaching resource. PMID- 15900053 TI - Rationale for using multiple antioxidants in protecting humans against low doses of ionizing radiation. AB - Health risks of low doses of ionizing radiation (10 cGy or less) may not be accurately estimated in humans by epidemiological study or mathematical modelling because of several inherent confounding factors including environmental, dietary and biological variables that cannot be accounted for in any radio epidemiological study. In addition, the expression of radiation-induced damage in humans not only depends upon total dose, dose rate, linear energy transfer (LET), and fractionation and protraction of total doses, but also on repair mechanisms, bystander effects, and exposure to chemical carcinogens, tumour promoters and other toxins. It also depends upon the levels of anti-carcinogenic and anti tumour promoting agents. Low doses of ionizing radiation should not be considered insignificant with regard to increasing the incidence of somatic mutations (neoplastic and non-neoplastic diseases) and heritable mutations in humans owing to its interaction with other toxins that can enhance damage produced by irradiation. It is very prudent to continue to support the well-established radiobiological concept that no radiation dose can be considered completely safe, and that all efforts must be made to reduce both the radiation dose and biological damage, no matter how small that damage might be, without sacrificing the benefits of radiation. Based on the results of many scientific experiments, formulations containing multiple antioxidants for biological protection against radiation damage in humans can be developed, and this strategy together with the existing physical concept of radiation protection, should further reduce potential risks of low doses of ionizing radiation in humans. PMID- 15900054 TI - Differences in bone mineral density and geometry in men and women: the Newcastle Thousand Families Study at 50 years old. AB - In 1947 Sir James Spence initiated the Newcastle Thousand Families study, which recruited all 1142 children born in the city between May and June that year. At the age of 50 years, 832 survivors were traced and invited to attend for measurement of bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA). The aim was to compare BMD measurements of men and women in this cohort, before and after adjustment for skeletal size. The femoral neck shaft angles (NSA) were also measured manually from the DXA scan printouts. A total of 171 men and 218 women agreed to participate. As expected men had greater bone mineral content and bone area at all sites (p<0.0001) and were taller and heavier (p<0.0001) than women. Men also had significantly higher BMD than women at all regions (p<0.0002), except at the femoral neck or lumbar spine. After correction for skeletal size and body weight, men had statistically significantly lower volumetric BMD at all sites. The measurement of NSA had good intra/interobserver errors and precision (coefficient of variations 0.79%, 1.2% and 1.2%). Men had significantly larger NSAs (mean 130 degrees , range 121-138 degrees ) than women (mean 128 degrees , range 119-137 degrees ). We conclude that there are gender differences in BMD, skeletal size and geometry in middle aged men and women, which together with the subsequent rate of bone loss, may influence fracture risk in later life. PMID- 15900055 TI - Acute ureteric calculus obstruction: unenhanced spiral CT versus HASTE MR urography and abdominal radiograph. AB - The aim of this study is to compare the performance of unenhanced spiral CT to the combination of HASTE MR urography (MRU) and plain abdominal radiography (KUB) in patients suspected of having acute calculus ureteric obstruction. 64 patients with suspected acute calculus ureteric obstruction were evaluated. The presence of perirenal fluid, presence and level of ureteric obstruction and calculi were assessed on both techniques. 44 of 64 (69%) patients had acute calculus ureteric obstruction based on clinical, radiographic or surgical findings. MRU showed perirenal fluid in acute ureteric obstruction (77%) with a greater sensitivity than CT showed stranding (45%). The combination of fluid and ureteric dilation on MRU showed a sensitivity of 93% (CT 80%), specificity of 95% (CT 85%), and accuracy of 94% (CT 81%). There were 61 findings of either fluid or ureteric dilatation on MRU in 44 acutely obstructed kidneys compared with 37 similar findings on CT (p<0.005). Although there was excellent reproducibility (Kappa=/>0.75) in the finding of perirenal fluid on MRU, there was only fair interobserver agreement (Kappa<0.4) regarding perirenal stranding on CT. MRU/KUB showed ureteric calculi in 21/29 (72%) of patients with calculi seen by CT. Overall, MRU/KUB revealed 2.4 abnormalities per acutely obstructed ureter compared with 1.8 abnormalities detected by CT. MRU/KUB using HASTE sequences can diagnose the presence of acute calculus ureteric obstruction with similar accuracy to spiral CT. The technique has less observer variability and is more accurate than CT in detecting evidence of obstruction such as perirenal fluid. PMID- 15900056 TI - Real-time blood-pool images of contrast enhanced ultrasound with Definity in the detection of tumour nodules in the liver. AB - Lower mechanical index (MI) technique with newer microbubble agents has been introduced into clinical practice as a newer ultrasound (US) imaging. However, the efficacy in detecting tumour nodules has not been proven scientifically. The aim of this study was to elucidate the efficacy of a blood-pool image of real time contrast-enhanced US under low MI in detecting liver tumours. 15 rabbits with VX-2 tumour were used; the number of implantations was none in two rabbits, one in four, two in five and three in four. US equipment was APLIO (Toshiba) with linear probe (3.5/7.0 MHz). The number, location and size of tumour nodules were examined by non-contrast tissue harmonic imaging (NC-US) or contrast-enhanced pulse subtraction harmonic imaging (C-US) under extra-low MI (MI 0.065) with the injection of Definity (30 microl kg(-1)). The number of tumour nodules detected by both NC-US and C-US were consistent with the histopathological results in five rabbits - two with none, two with one nodule and one with two nodules. In the other 10 rabbits, C-US showed all the implanted tumours and small daughter nodules around them that were confirmed by histopathology. However, NC-US failed to demonstrate two implanted nodules and all the daughter nodules. On the basis of the histopathological results, detectability of implanted tumour was not significantly different between NC-US (24/26, 92.3%) and C-US (26/26, 100%). However C-US was superior to NC-US in delineating the nodules and in detecting small daughter nodules. The sizes of the implanted tumour nodules measured by histopathology correlated closely with those measured by C-US. Real-time blood pool images by pulse subtraction harmonic imaging under extra-low MI with Definity will contribute to the improvement of the ultrasound delineation and detection of liver tumours. PMID- 15900057 TI - Effectiveness of flexible noise control image processing for digital portal images using computed radiography. AB - The purpose of this study was to evaluate the clinical efficacy of flexible noise control (FNC) image processing in off-line computed radiography (CR) portal images. An observer study was designed to compare FNC with multiobjective frequency processing (MFP) in images acquired using a CR portal imaging device (CRPID). The image processing of MFP and FNC used the same data and required no additional irradiation of patients, and all images were printed on 10-bit grey level dry laser film. Four radiation technologists and one oncologist served as observers and evaluated 40 sets of images for three different treatment sites: brain, lung and pelvis. Six to 10 anatomical landmarks were selected from each treatment site. Each observer was asked to rate each landmark in terms of its clinical visibility and rate the ease of making the pertinent verification in the MFP- and FNC-processed images. In the ratings of the visibility of landmarks and for the verification of treatment ports, FNC-processed images were more visible than MFP-processed images except for several landmarks in the anteroposterior (AP) pelvis such as the pubic symphysis. The visibility of landmarks in FNC processed images was comparable with that in MFP-processed images. The verification of treatment ports using the CRPID with FNC was generally achievable. In conclusion, this study suggests that FNC is effective for image processing of CR portal images. PMID- 15900058 TI - Comparative analyses of the dynamic properties of the bladder wall studied by repetitive pelvic CT scans of patients and cryo-sections of cadavers. AB - In radiotherapy planning systems, delineation of hollow normal tissue organs, such as the bladder, is time-consuming. Automated delineation may presuppose two assumptions: (1) the bladder resembles a spherical shell and (2) the volume of bladder tissue is preserved regardless of the volume of urine (luminal volume) inside. The purpose of the present study was to test these assumptions. 22 CT scans from 7 patients were studied retrospectively. Transverse cross-sectional areas enclosed by the outer contour (A(out)) and inner contour of the bladder (A(in)) were recorded from the images. Hence, the transverse cross-sectional area of the wall, A(wall)=A(out)-A(in), and the volume of bladder tissue at various luminal volumes, could be calculated. To quantify the method uncertainty, the same procedure was applied on three spherical plastic phantoms. The results were also compared with data from the Visible Human Project's photographs of cadaver cryo-sections. Assumption no. 1 stated above, implies that A(wall) is constant regardless of the level of intersection of the sphere. The data from cryo sections revealed a positive correlation for A(wall) and A(out), in contradiction to assumption no. 1 (p<0.001). The corresponding association derived from the repetitive CT scans of patients was also statistically significant (p<0.001) although linear regression revealed a less steep slope. A relationship was found between the volume of bladder tissue and luminal volume, hence contradicting assumption no. 2 (p<0.001). In conclusion the cross-sectional wall areas of the bladder, measured from patient CT scans, increase slightly with luminal cross sectional areas in contradiction to expected values derived from a simplistic spherical shell model. In addition, the volume of bladder tissue is related to the luminal volume. Our results may be of practical value when developing automated delineation tools in radiotherapy planning systems. PMID- 15900059 TI - Differentiation between solitary brain metastasis and high-grade glioma by diffusion tensor imaging. AB - We tested our hypothesis that fractional anisotropy (FA) maps of diffusion tensor imaging could be used to differentiate between a solitary brain metastasis and a high-grade glioma. In seven patients with a solitary metastasis and seven patients with a high-grade glioma, FA values of enhancing and non-enhancing parts of the tumour were compared. Additionally, we visually assessed FA maps. No significant difference in the FA values of either the enhancing or non-enhancing part was found between the two groups. In the visual assessment, displacement of subcortical white-matter fibres was found in five of the seven metastasis patients, but in only one glioma patient. Additionally, discrimination between tumour and oedema was possible in three of the seven metastasis patients, but not in any glioma patient. Although FA values are not helpful in differentiating between the two groups, visual differences in FA values can allow the differentiation. Displacement of white-matter fibres is another finding suggestive of metastasis. PMID- 15900060 TI - The effect of introducing radiographer reporting on the availability of reports for Accident and Emergency and General Practitioner examinations: a time-series analysis. AB - This study explores whether the introduction of selectively trained radiographers reporting Accident and Emergency (A&E) X-ray examinations of the appendicular skeleton affected the availability of reports for A&E and General Practitioner (GP) examinations at a typical district general hospital. This was achieved by analysing monthly data on A&E and GP examinations for 1993-1997 using structural time-series models. Parameters to capture stochastic seasonal effects and stochastic time trends were included in the models. The main outcome measures were changes in the number, proportion and timeliness of A&E and GP examinations reported. Radiographer reporting X-ray examinations requested by A&E was associated with a 12% (p=0.050) increase in the number of A&E examinations reported and a 37% (pA transition (NDOR1*1), detected in 24/200 Caucasian and 1/49 Japanese individuals, producing a valine to isoleucine substitution at codon 522 in the NADPH binding region. Expression of the flavin adenine dinucleotide/reduced nicotinamide phosphate dehydrogenase (NADPH) domain in Escherichia coli showed a significant 74% reduction in potassium ferricyanide reductase activity, but no effect on NADPH binding. NDOR1_v1 showed a 10-fold decrease in affinity for NADPH, and a 90% reduction in ferricyanide reductase activity. CONCLUSIONS: We have discovered a polymorphic variant of NDOR1, NDOR1*1, that produces a functionally impaired enzyme. This will help define the structure and function of NDOR1 and its relationship to cancer and other diseases. PMID- 15900211 TI - The A1 allele of the human D2 dopamine receptor gene is associated with increased activity of striatal L-amino acid decarboxylase in healthy subjects. AB - The A1 allele of the TaqI restriction fragment length polymorphism (RFLP) of the human dopamine D2 receptor gene (DRD2) is associated with a low density of D2 dopamine receptors in the striatum. Because of the important role of D2 autoreceptors in regulating dopamine synthesis, we aimed to examine whether subjects with the A1 allele have altered presynaptic dopamine function in the brain. We also studied the effects of two other DRD2 polymorphisms, C957 T and- 141C Ins/Del, which have been suggested to affect D2 receptor levels in brain. The relationships between the TaqIA RFLP, C957 T and--141C Ins/Del polymorphisms and striatal dopamine synthesis in 33 healthy Finnish volunteers were studied using positron emission tomography and [18F]fluorodopa ([18F]FDOPA), a radiolabelled analog of the dopamine precursor L-DOPA. Heterozygous carriers of the A1 allele (A1/A2; 10 subjects) had significantly higher (18%) [18F]FDOPA uptake in the putamen than subjects without the A1 allele (A2/A2; 23 subjects). C957 T and--141C Ins/Del polymorphisms did not significantly affect [18F]FDOPA Ki values. These results demonstrate that the A1 allele of DRD2 gene is associated with increased striatal activity of aromatic L-amino acid decarboxylase, the final enzyme in the biosynthesis of dopamine and the rate-limiting enzyme for trace amine (e.g. beta-phenylethylamine) synthesis. The finding can be explained by lower D2 receptor expression leading to decreased autoreceptor function, and suggests that dopamine and/or trace amine synthesis rate is increased in the brains of A1 allele carriers. PMID- 15900212 TI - Association of catechol-O-methyltransferase with smoking cessation in two independent studies of women. AB - OBJECTIVES AND METHODS: The Val108/158Met polymorphism in the gene that encodes COMT, a dopamine metabolizing enzyme, results in a three- to four-fold reduction in COMT activity. To determine if the lower activity Met allele of COMT was associated with smoking cessation in women, we used two independent studies: a population-based case--control study and a nicotine replacement clinical trial. RESULTS: In the case--control study, women with two Met alleles were significantly more likely to be ex-smokers than current smokers [OR=1.82, 95% CI (1.05, 3.17), P=0.03]. In the nicotine replacement clinical trial, among women, the Met/Met genotype was associated with a higher probability of smoking cessation based on both point prevalence and prolonged abstinence outcomes [OR=2.96, 95% CI (1.07, 8.14), P=0.04; OR=3.23, 95% CI (1.13, 9.20), P=0.03, respectively]. CONCLUSIONS: This first report of a significant association between COMT Val108/158Met and smoking cessation suggests that COMT variation has an effect on smoking behavior in women. PMID- 15900213 TI - Genetic variation in the bleomycin hydrolase gene and bleomycin-induced pulmonary toxicity in germ cell cancer patients. AB - OBJECTIVE: Use of bleomycin as a cytotoxic agent is limited by its pulmonary toxicity. Bleomycin is mainly excreted by the kidneys, but can also be inactivated by bleomycin hydrolase (BMH). An 1450A>G polymorphic site in the BMH gene results in an amino acid substitution in the C-terminal domain of the protein. Deletion of this domain, including the polymorphic site, reduces enzymatic activity. We investigated the relation between the BMH genotype and the risk of bleomycin-induced pneumonitis (BIP). METHODS: From male germ cell cancer patients, treated with bleomycin-containing chemotherapy at the University Hospital Groningen, The Netherlands, between 1977 and 2003, data were collected on age, cumulative bleomycin dose, pretreatment creatinine clearance, pulmonary metastases, lung function parameters, and occurrence of BIP. BIP was defined as: death due to BIP, or presence of clinical and/or radiographic signs of BIP during or following treatment. Polymerase chain reaction and restriction fragment length polymorphism were used to determine the BMH genotype. RESULTS: BIP developed in 38 (11%) of 340 patients; four of these cases were fatal. BMH genotype distribution did not differ between patients with and those without BIP. Patients with BIP were older and had a lower pretreatment creatinine clearance. Changes in pulmonary function tests were similar in patients with different genotypes. CONCLUSIONS: The BMH genotype was not associated with the development of BIP nor with changes in pulmonary function tests. Since renal function is important for bleomycin pharmacokinetics, variations in renal clearance may have obscured significant effects of the BMH genotype. PMID- 15900214 TI - Variation in the alpha2B-adrenergic receptor gene (ADRA2B) and its relationship to vascular response in vivo. AB - The alpha2B-adrenergic receptor (ADRA2B) plays an important role in vasoconstriction and blood pressure regulation. One common variant in the ADRA2B gene (del 301--303) has been identified, and results in markedly decreased receptor desensitization in vitro but does not alter vascular sensitivity in vivo. Therefore, we fully characterized genetic variations in ADRA2B and related them to phenotype in vivo. We examined 5812 bp of contiguous sequence of ADRA2B (promoter, exonic, and 3'-untranslated region; 3'-UTR) using the polymerase chain reaction to amplify the genomic target followed by bidirectional sequencing (n=68). Haplotypes were inferred using an expectation maximization algorithm. Vasoconstriction in response to increasing doses of the highly selective alpha2 adrenergic receptor agonist, dexmedetomidine (0.01--1000 ng/min) was measured in the dorsal hand vein using a linear variable differential transformer. The dose that produced 50% (ED50) of maximum venoconstriction (Emax) was determined for each subject from the individual dose--response curves. ED50 and Emax were compared in subjects with and without variant alleles and haplotypes of interest. We identified 24 variable sites, 12 in the promoter region, five in the coding region (including two previously described as non-synonymous variants) and seven in the 3'-UTR region. Four haplotypes were inferred, representing approximately 95% of the cohort. One haplotype, characterized by two single nucleotide polymorphisms in the promoter region, and one in the 3'-UTR, occurred in seven of 38 African-Americans, and was associated with a lower Emax, 61.3% [95% confidence interval (CI) 39.5--83.0, n=7] compared to 78.1% (CI 73.8--82.5) in wild-types (n=61) (P=0.02). There was no association between the nine common variants and dexmedetomidine ED50. We have described novel variants and haplotypes of the ADRA2B gene. These do not alter sensitivity to a selective alpha2-adrenergic receptor agonist but some may decrease maximal venoconstriction in vivo. PMID- 15900215 TI - Relative impact of CYP3A genotype and concomitant medication on the severity of atorvastatin-induced muscle damage. AB - Atorvastatin is metabolized through enzymes encoded by members of the cytochrome P-450 (CYP) 3A gene family. Some patients who take atorvastatin along with concomitant medications known to inhibit CYP3A enzyme activity (e.g. itraconazole) develop rhabdomyolysis secondary to a severe drug-induced myopathy. The present study aimed to characterize the relationship between CYP3A gene polymorphisms and atorvastatin-induced muscle damage in the context of concomitant medication. The study employed a retrospective case--control (n=137) design. Study subjects were recruited from the general patient population served by Marshfield Clinic, a large horizontally integrated multispecialty group practice located in central Wisconsin, and case assignment was based upon both subjective (myalgia) and objective inclusion criteria [elevated serum creatine kinase (CK) levels]. The primary outcome was the relationship between serum CK level and CYP3A genotype. CYP3A genotype was not associated with an increased risk for the development of atorvastatin-induced muscle damage. CYP3A4*1B and CYP3A5*3 allele frequencies were similar in cases (n=68) and controls (n=69). Conversely, CYP3A genotype was associated with an increased severity of atorvastatin-induced muscle damage. An association was identified between the non functional CYP3A5*3 allele and the magnitude of serum CK elevation in case patients experiencing myalgia. Patients who were homozygous for CYP3A5*3 demonstrated greater serum CK levels than patients who were heterozygous for CYP3A5*3, when concomitant lipid-lowering agents were sequentially removed from the analysis (P=0.025 without gemfibrozil, P=0.010 without gemfibrozil and niacin). The study demonstrates that patients who develop myalgia while taking atorvastatin are more likely to experience a greater degree of muscle damage if they express two copies of CYP3A5*3. PMID- 15900216 TI - Glutathione S-transferase polymorphisms and susceptibility to neuroblastoma. AB - There is evidence to suggest that polymorphic variations in the glutathione S transferase (GSTs) are associated with cancer susceptibility. The GST supergene family includes several genes with well characterized polymorphisms. Approximately 50% of the Caucasian population is homozygous for deletions in GSTM1 and approximately 20% are homozygous for deletions in GSTT1. Deletions lead to an absence of the protein, thus resulting in conjugation deficiency of mutagenic electrophiles to glutathione. The GSTP1 gene displays a polymorphism at codon 105 resulting in an Ile to Val substitution, which alters the enzymatic activity of the protein, and this has been suggested as a putative high-risk genotype in various cancers. In the present study, we investigated the relationship between GSTs polymorphism and the susceptibility to neuroblastoma, comparing GSTs genotypes of 256 children with neuroblastoma with those of 392 normal control subjects. No significant differences of allele frequencies were found between patients and controls. Within the neuroblastoma group, we further investigated whether any particular GSTs genotype was correlated with clinical and biological characteristics at diagnosis, but no association was detected. Our data do not support an important effect of GSTs genotype on neuroblastoma susceptibility. PMID- 15900217 TI - Polymorphisms in the mitochondrial aldehyde dehydrogenase gene (Aldh2) determine peak blood acetaldehyde levels and voluntary ethanol consumption in rats. AB - Dependence on alcohol, a most widely used drug, has a heritability of 50-60%. Wistar-derived rats selectively bred as low-alcohol consumers for many generations present an allele (Aldh2(2)) of mitochondrial aldehyde dehydrogenase that does not exist in high-alcohol consumers, which mostly carry the Aldh2(1) allele. The enzyme coded by Aldh2(2) has a four- to five-fold lower affinity for NAD than that coded by Aldh2(1). The present study was designed to determine whether these polymorphisms account for differences in voluntary ethanol intake and to investigate the biological mechanisms involved. Low-drinker F0 Aldh2(2)/Aldh2(2) rats were crossed with high-drinker F0 Aldh2(1)/Aldh2(1) rats to obtain an F1 generation, which was intercrossed to obtain an F2 generation that segregates the Aldh2 alleles from other genes that may have been coselected in the breeding for each phenotype. Data show that, with a mixed genetic background, F2 Aldh2(1)/Aldh2(1) rats voluntarily consume 65% more alcohol (P<0.01) than F2 Aldh2(2)/Aldh2(2) rats. A major phenotypic difference was a five fold higher (P<0.0025) peak blood acetaldehyde level following ethanol administration in the lower drinker F2 Aldh2(2)/Aldh2(2) compared to the higher drinker F2 Aldh2(1)/Aldh2(1) animals, despite the existence of identical steady state levels of blood acetaldehyde in animals of both genotypes. Polymorphisms in Aldh2 play an important role in: (i) determining peak blood acetaldehyde levels and (ii) modulating voluntary ethanol consumption. We postulate that the markedly higher peak of blood acetaldehyde generated in Aldh2(2)/Aldh2(2)(2) animals is aversive, leading to a reduced alcohol intake in Aldh2(2)/Aldh2(2) versus that in Aldh2(1)/Aldh2(1) animals. PMID- 15900218 TI - Phenotyping-genotyping of alternatively spliced genes in one step: study of CYP3A5*3 polymorphism. AB - Alternative splicing is required to increase the mRNA diversity of many genes, but can also be responsible for the abnormal expression of genes. For example, the CYP3A5*3 defective allele is caused by a single nucleotide polymorphism in intron 3. This mutation activates a cryptic acceptor splice site, which leads to the insertion of an intronic sequence containing premature termination codons in the mature mRNA, and hence the very low CYP3A5 protein expression in 75% of the Caucasian population. In the present study, we propose a novel strategy based on the quantitative real-time polymerase chain reaction with SYBR Green I chemistry, followed by melting curve analysis, to demonstrate and quantify the amount of splice variant mRNA. Using oligonucleotides flanking the insertion site, two products with different size can be obtained, which can be resolved by melting curve analysis. The relative ratio of differently spliced RNA can be estimated at the plateau phase by using the peak height ratio. For the CYP3A5 gene, the genotype, the level of expression and the proportion of alternatively spliced products were determined in a single reaction without DNA sequencing. PMID- 15900219 TI - Paraoxonase-1 promoter polymorphism C--107T and serum apolipoprotein AI interact to modulate serum paraoxonase-1 status. AB - OBJECTIVES: The objective was to examine the hypothesis that modifications to paraoxonase-1 specific activity (SP, activity per unit mass peptide) could contribute to serum paraoxonase-1 status, a determinant of the clinical efficacy of the enzyme. METHODS: Enzyme activities and concentrations were determined in a large population (n=912) of patients and controls. SP were subsequently examined as a function of paraoxonase-1 gene polymorphisms, plasma lipids and lipoproteins, and physiological and pathophysiological parameters. RESULTS: Pathophysiological parameters (diabetes, metabolic syndrome, smoking, aging) did not promote variations in paraoxonase-1 SP, whilst coronary disease lowered SP (P<0.003). No serum lipid, apolipoprotein or lipoprotein component had an impact on specific activity, with the exception of apolipoprotein AI (P<0.005, both substrates). The paraoxonase-1 promoter C--107T and Q192R polymorphisms influenced SP and, together with apolipoprotein AI, were highly significant, independent determinants in regression models. There was an interaction between apolipoprotein AI and the C--107T polymorphism, which significantly modulated SP and serum paraoxonase-1 status. CONCLUSIONS: Enzyme inactivation giving rise to modulated activity per unit mass of peptide is not a major contributor to pathological effects of disease on serum paraoxonase-1 status. The C--107T polymorphism and serum apolipoprotein AI have major impacts individually on SP and also provide an example of gene-environment interaction to modulate such activities. These effects accentuate the differences between--107C and--107T allele carriers in terms of serum paraoxonase-1 status. The data underline the complexity of the factors that determine serum paraoxonase-1 status and suggest that the latter would benefit from therapeutic modulation of serum high density lipoproteins. PMID- 15900220 TI - Molecular psychiatry meeting, February 2004, Park City, Utah. AB - The 11th Annual Molecular Psychiatry Meeting was held from 8 to 10 February 2004, at the Treasure Mountain Inn in Park City, Utah. The meeting was hosted by Bill Byerley, University of California, Irvine. PMID- 15900221 TI - Polymorphisms in the brain-derived neurotrophic factor gene are not associated with either anorexia nervosa or schizophrenia in Dutch patients. PMID- 15900222 TI - Genetic linkage analysis of the X chromosome in autism, with emphasis on the fragile X region. AB - The higher prevalence of autism in males than in females suggests the possible involvement of the X chromosome. To test the hypothesis that there are mutations increasing susceptibility to autism on the X chromosome, and in particular the distal portion of the long arm that encompasses the FMRI and MECP2 loci, a genetic linkage study was performed. Twenty-two fragile X-negative families multiplex for autism and related disorders were used for the study. Linkage analysis, for markers in the Xq27-q28 region, using model-free likelihood-based analysis, produced a maximum MLOD of 1.7 for the narrowest diagnostic category of the typical autism/severe autism spectrum, and nonparametric analysis produced a maximum non-parametric lod (NPL) score of 2.1 for a broad phenotype diagnostic model. Thus, this study offers modest support for a susceptibility locus for autism within the Xq27-q28 region. Further genetic investigations of this region are warranted. PMID- 15900223 TI - Characterization of CTG/CAG repeats on chromosome 18: a study of bipolar disorder. AB - Anticipation has been frequently found in bipolar families ascertained for linkage studies. An association of polymorphic triplet repeats with the bipolar phenotype in some pedigrees has been proposed. We have previously found linkage to chromosome 18 in a set of families with evidence of anticipation. As part of a search for CAG/CTG motifs on chromosome 18, we screened a genomic chromosome 18 cosmid library and identified 65 loci with trinucleotide repeats. Eleven of 33 genotyped loci were polymorphic, though none of these showed any evidence of instability. We performed genetic analysis of six loci in the Hopkins/Dana bipolar pedigrees ascertained for a genetic linkage study of bipolar disorder and found that the CAG repeat within the AD4D2 clone on 18q21.1 showed nominally significant over-transmission of the rare CAG23 allele (P=0.034). We have characterized all 65 trinucleotide repeats and flanking sequences with GENSCAN analysis and find that 29 were predicted to be in coding regions. These 29 trinucleotide-repeat-containing genes may be involved in functional modulation of their respective proteins, and may be candidates for other diseases or disease mechanisms that map to this region. PMID- 15900224 TI - Heritability of substance dependence in a native American population. AB - OBJECTIVES: To estimate the heritability of substance dependence and associated symptoms in a sample of Southwest California (Mission) Indians. METHODS: Families from eight contiguous Indian reservations were recruited in order to ascertain information on substance dependence symptoms and diagnoses using a semi structured diagnostic interview. Dependence diagnoses for alcohol, marijuana, stimulants and a measure of regular tobacco usage, any drug dependence or tobacco usage were obtained. Composite measures of alcohol dependence symptoms for withdrawal, drinking severity, antisocial problems and interpersonal problems were constructed from the nine groups of symptoms summarized in the diagnostic interview. Heritability estimates were calculated using variance component methods, as implemented in SOLAR. RESULTS: In this population, marijuana dependence (0.38) and regular tobacco use (0.43), alcohol dependence (DSM-III-R 0.19; ICD-10, 0.29) and stimulant dependence (0.25) showed evidence for moderate genetic influences as determined by heritability estimates. Four phenotypes constructed using the composite symptoms of alcohol dependence revealed that withdrawal had the highest heritability estimate (0.71), followed by antisocial problems (0.36) and drinking severity (0.34). Symptom clusters reflecting interpersonal problems did not appear to be highly heritable (0.19). CONCLUSION: Marijuana dependence, regular tobacco usage and composite phenotypes constructed from alcohol dependence symptoms for antisocial problems, drinking severity and withdrawal generally have patterns of familial aggregation, suggesting that they can be successfully used for linkage analysis in this Southwest California Indian sample. PMID- 15900225 TI - Association between the COMT Val158Met polymorphism and propensity to anxiety in an Australian population-based longitudinal study of adolescent health. AB - OBJECTIVES: Catechol-O-methyltransferase plays a central role in the metabolism of biogenic amines such as norepinephrine, dopamine and serotonin. Functional studies have demonstrated a dose relationship between ValMet genotypes and catechol-O-methyltransferase activity. Compared with the ValVal genotype, the ValMet and MetMet genotypes result in two- and four-fold reductions in catechol-O methyltransferase activity, respectively. Two recent reports have observed the association between the MetMet genotype and risk of anxiety in adult populations. We examined the association between the ValMet genotypes and propensity to anxiety across adolescence. METHODS: Participants were drawn from an eight-wave study of the mental and behavioural health of over 2000 young Australians followed from 14 to 24 years of age (Victorian Adolescent Health Cohort Study, 1992 to present). DNA was received from 962 participants using a cheek swab collection method. RESULTS: The odds of reporting persistent episodic anxiety (phobic avoidance, panic attacks) were doubled among carriers of the MetMet genotype (odds ratio 2.0, 95% confidence interval 1.1-3.4, P=0.014). A dose relationship between additional copies of the Met allele and persistent episodic anxiety was also observed (1.5, 1.1-1.94, P=0.007). Stratification by sex showed that the risk effect of the Met allele was among females only. No association was observed for measures of neuroticism, persistent generalized anxiety, or a composite measure of psychiatric distress. CONCLUSION: These data replicate previous findings suggesting association between the ValMet polymorphism and specific expressions of anxiety among females. PMID- 15900226 TI - Paternal age and intelligence: implications for age-related genomic changes in male germ cells. AB - BACKGROUND: A robust association between advancing paternal age and schizophrenia risk is reported, and genetic changes in the germ cells of older men are presumed to underlie the effect. If that is so, then the pathway may include effects on cognition, as those with premorbid schizophrenia are reported to have lower intelligence. There are also substantial genetic influences on intelligence, so de novo genetic events in male germ cells, which accompany advancing paternal age, may plausibly influence offspring intelligence. OBJECTIVE: An association of paternal age with IQ in healthy adolescents may illuminate the mechanisms that link it to schizophrenia. METHOD: We examined the association of paternal age and IQ scores using the Israeli Army Board data on 44 175 individuals from a richly described birth cohort, along with maternal age and other potential modifiers. RESULTS: A significant inverted U-shaped relationship was observed between paternal age and IQ scores, which was independent from a similar association of IQ scores with maternal age. These relationships were not significantly attenuated by controlling for multiple possible confounding factors, including the other parent's age, parental education, social class, sex and birth order, birth weight and birth complications. Overall, parental age accounted for approximately 2% of the total variance in IQ scores, with later paternal age lowering non-verbal IQ scores more than verbal IQ scores. CONCLUSION: We found independent effects of maternal and paternal age on offspring IQ scores. The paternal age effect may be explained by de novo mutations or abnormal methylation of paternally imprinted genes, whereas maternal age may affect fetal neurodevelopment through age-related alterations in the in-utero environment. The influence of late paternal age to modify non-verbal IQ may be related to the pathways that increase the risk for schizophrenia in the offspring of older fathers. PMID- 15900227 TI - Association of schizophrenia in African Americans to polymorphism in synapsin III gene. AB - Linkage studies in families with schizophrenia have pointed to chromosome 22q12 q13 as one of several regions of the genome that may contain a susceptibility gene. The gene coding for synapsin III, an intrinsic synaptic vesicle membrane protein, maps to this target region. Two tightly linked single-nucleotide polymorphisms were recently found in a small subset of patients with SZ - a synonymous variant, L469L (469G>A), and a non-synonymous variant, S470N (470G>A) which results in the loss of a mitogen-activated protein kinase serine phosphorylation site. We also found a slight increase in 470A in Caucasian patients from the US with schizophrenia. But, the sample size and allele frequency were too small to draw definitive conclusions. However, both single nucleotide polymorphisms were much more polymorphic in African American controls than in Caucasian controls, thereby providing a better sample cohort to analyze for schizophrenia involvement. For the codon 469 single-nucleotide polymorphisms, a 50-fold increase was observed in the frequency of 469A in African Americans compared with Caucasians. Furthermore, there was an increase in the percentage of African American patients with schizophrenia who were homozygous for the 469A allele compared with controls who were homozygous (11 versus 5%; AA vs. all other genotypes - Fisher statistic=3.08, P=0.04, one-tailed). An increase in 470A heterozygotes was also found, but the results fell short of being statistically significant. The findings support a role for synapsin III in a subset of African American patients with schizophrenia and raises questions about selective pressure in Africa to account for the extraordinary disparity of the 469 and 470 single-nucleotide polymorphisms in different ethnic populations. PMID- 15900228 TI - Novelty seeking: interaction between parental alcohol use and dopamine D4 receptor gene exon III polymorphism over 17 years. AB - Recent meta-analyses have questioned the association between the dopamine receptor D4 (DRD4) gene polymorphism and the temperament trait of novelty seeking, and proposed an interaction between the polymorphism and other factors. We wanted to test whether parental alcohol use during childhood moderated the effect of an offspring dopamine receptor gene (DRD4) polymorphism on the temperament trait of novelty seeking in adulthood. A population-based sample of children and adolescents (n=2149) and their parents was examined in 1980 and 1983 on parental alcohol use and rearing practices. In 1997, study participants completed the Temperament and Character Inventory for the novelty-seeking temperament trait, and a subsample (n=150) was genotyped for the DRD4 exon III polymorphism. For the participants with the father, but not the mother, reporting more frequent alcohol consumption or drunkenness in examinations 17 and/or 14 years before the novelty-seeking assessment, an association between the short (two- or five-repeat) alleles of the DRD4 gene and extremely high novelty-seeking scores was observed. When the father reported less frequent alcohol consumption or drunkenness, the genotype was not associated with novelty seeking. The association remained after controlling for sex, age, and maternal child-rearing. These results provide preliminary information on gene-environment interaction on the temperament trait of novelty seeking and may partly explain the heterogeneity of findings concerning the association between DRD4 polymorphisms and novelty seeking. PMID- 15900229 TI - Association of MAO A polymorphism and alcoholism in Brazilian females. AB - Among the different possible genes involved in the alcoholism etiology, the X linked monoamine oxidase A gene is a good candidate. The aim of this study was to assess whether a functional VNTR polymorphism in the promoter region of the monoamine oxidase A gene is associated with alcoholism, comparing patients of both sexes. Ninety-three alcohol-dependent patients (51 males, 42 females) and 93 sex-matched normal controls were engaged. In the total sample, the genotype containing at least one three-repeat allele was significantly more frequent among alcohol-dependent patients than controls (P=0.01). However, when the two sexes were analyzed separately, the difference was statistically significant only for females. This is of particular interest as rates of alcoholism in Brazil are markedly lower in females. Our results suggest that this monoamine oxidase A polymorphism could play a role in susceptibility to alcoholism, which may differ across sexes. PMID- 15900230 TI - Reward dependence is related to norepinephrine transporter T-182C gene polymorphism in a Korean population. AB - It is well established that approximately 50% of the variance in personality traits is genetic. The goal of this study was to investigate a relationship between personality traits and the T-182C polymorphism in the norepinephrine transporter gene. The participants included 115 healthy adults with no history of psychiatric disorders and other physical illness during the past 6 months. All participants were tested with the Temperament and Character Inventory and genotyped norepinephrine transporter gene polymorphism. Differences on the Temperament and Character Inventory dimensions among three groups were examined with one-way analysis of variance. Our study suggests that the norepinephrine transporter T-182C gene polymorphism is associated with reward dependence in Koreans, but the small number of study participants and their sex and age heterogeneity limits generalization of our results. Further studies are necessary with a larger number of homogeneous participants to confirm whether the norepinephrine transporter gene is related to personality traits. PMID- 15900231 TI - Suggestive evidence for association of D2S2188 marker (2q31.1) with autism in 143 Sicilian (Italian) TRIO families. AB - We have screened 143 Sicilian (Italian) families with one autistic child to verify, by a linkage disequilibrium approach, the involvement of the 2q31.1 region in the cause of the disease in these families. Our study design includes the use of intrafamilial association to prevent a population stratification bias and ethnic homogeneity of the sample. The results of our analysis provided suggestive evidence of the occurrence of transmission disequilibrium between autism and the D2S2188 polymorphism in Sicilian TRIO families, a finding which provides further and independent support to the hypothesis of the existence of a susceptibility gene (or genes) for autism on chromosome 2q. PMID- 15900232 TI - Association study of catechol-O-methyltransferase gene polymorphism in Korean male alcoholics. AB - OBJECTIVE: Catechol-O-methyltransferase, which has a functional genetic polymorphism, plays an important role in dopamine metabolism. The study analyzed the association between the catechol-O-methyltransferase gene polymorphism and alcohol dependence in the Korean population. METHODS: Ninety-seven male alcoholics and 94 male age-matched normal controls were enrolled in this study. Polymerase chain reaction-based genotyping was used to verify the presence of the catechol-O-methyltransferase gene polymorphism. The patients were divided into two subtypes (violent group and non-violent group) according to their history of violent behavior. RESULTS: No difference in the distribution of the catechol-O methyltransferase genotypes (H/H, H/L, L/L) and alleles (H, L) was observed between the patients and the controls. However, the differences between the violent and non-violent groups were significant in terms of the frequencies of the COMT genotypes (chi=7.977, df=2, P=0.019) and the alleles (chi=6.832, df=1, P=0.012). In addition, significant differences in the frequencies of the catechol O-methyltransferase allele (chi=4.481, df=1, P=0.040) were observed between the non-violent group and the controls. CONCLUSIONS: This suggests that the catechol O-methyltransferase gene polymorphism is not associated with the development of alcohol dependence, but may affect the susceptibility to a clinical heterogeneity of alcohol dependence, at least in the Korean population. PMID- 15900233 TI - Na+/H+ exchanger inhibitor prevented endothelial dysfunction induced by high glucose. AB - The aims of this study were to examine whether cariporide, a selective Na+/H+ exchanger inhibitor, has protective effects against endothelial dysfunction induced by high glucose in vitro and to investigate the potential mechanisms. Exposure of rat aorta rings to high glucose (44 mmol/L) for 6 hours caused an inhibition of acetylcholine-induced endothelium-dependent relaxation but had no effect on sodium nitroprusside-induced endothelium-independent relaxation. Treatment with cariporide (0.01, 0.1, 1 micromol/L) of aortic rings incubated with high-glucose medium attenuated the impaired endothelium-dependent relaxation in a dose-dependent manner. Furthermore, high glucose resulted in an increase of malondialdehyde and a decrease of superoxide dismutase activity in rat aorta rings, and these effects were reversed by cariporide. In addition, cariporide was able to inhibit the activation of Na+/H+ exchanger induced by high glucose in cultured endothelial cells. These findings suggest that the endothelial dysfunction induced by high glucose in vitro is caused by the activation of Na+/H+ exchanger. PMID- 15900234 TI - Evaluation of the coronary artery bypass graft. PMID- 15900235 TI - The current status of sentinel lymph node mapping in non-small cell lung cancer. AB - Sentinel lymph node (SLN) mapping has become a common procedure in the treatment of breast cancer and malignant melanoma. Its primary benefit is that it enables surgeons to avoid nontherapeutic lymph node dissection and the complications that follow. There are also several studies of the use of SLN mapping in the treatment of non-small cell lung cancer (NSCLC) reported in the English literature, and all present evidence for the existence of SLNs in NSCLC. Nevertheless, SLN mapping is not widely used in the treatment of NSCLC for several reasons: first, special precautions are required to minimize exposure when radioisotopes are used as tracers; second, it is difficult to detect the blue dyes used as tracers within anthoracotic thoracic lymph nodes; and third, major complications comparable to the arm edema seen in breast cancer or the lymphedema and nerve injury seen in melanoma are not seen with mediastinal lymph node dissection (MLND). To address these issues, new techniques are being developed by groups at several institutes, including our own. We believe that SLN mapping will enable surgeons to more precisely stage NSCLC, after which more sensitive techniques can be employed on a limited amount of tissue to detect occult micrometastatic disease with less cost and effort. SLN mapping can also be applied to video-assisted thoracic surgery (VATS) for NSCLC, enabling surgeons to avoid nontherapeutic and technically difficult MLND often necessary with traditional open surgery. For all of these reasons, we think that SLN mapping will be useful in the treatment of NSCLC, and that further development aimed at making SLN mapping a practical surgical procedure is warranted. PMID- 15900236 TI - Physiologic aspects in human lung transplantation. AB - Heart-lung transplantation (HLT), followed by single lung transplantation (SLT) and subsequently bilateral lung transplantation (BLT) have been developed as treatments for patients with end-stage pulmonary diseases. Initially, SLT was limited to idiopathic pulmonary fibrosis (IPF) cases and thought to be contraindicated not only for infectious diseases, but also for non-infectious diseases, including pulmonary emphysema (PE) and primary pulmonary hypertension (PPH), based on physiologic points of view. However, SLT is now widely performed for those non-infectious diseases and most of the recipients return to a normal active life. It is quite possible that BLT is superior to SLT in terms of pulmonary function, and it has been reported that BLT is better for PE and PPH patients in regards to perioperative course, postoperative exercise capacity, and long-term survival. For those situations and because of the present scarcity of donor organs, SLT must be utilized for selected non-infectious diseases for which it is safe and effective. When a single lung is replaced for IPF, PE, and PPH recipients, different physiologic situations are produced postoperatively, the understanding of which is extremely important to achieve good results, not only in the perioperative but also in the long term. PMID- 15900237 TI - Usefulness of automatic triangular anastomosis for esophageal cancer surgery using a linear stapler (TA-30). AB - In a clinical evaluation, thirty-one patients with thoracic esophageal cancer who were treated from 1997 to 2000 were selected as subjects including those who underwent hand sewn anastomosis method, circular stapler method and triangular anastomosis method. Incidence of anastomosis failure was 27.3%, 25.0% and 8.3% for the hand sewn anastomosis method, circular stapler method and triangular anastomosis method, respectively. Anastomosis stenosis was found in 32.4%, 45.6% and 8.3% for the hand sewn anastomosis method, circular stapler method and triangular anastomosis method, respectively. In the basic examination, reduction rate of anastomosis caliber was 22.1+/-4.8%, 14.9+/-1.4% and 7.37+/-0.9%, for the hand sewn anastomosis method, circular stapler method and triangular anastomosis method. Microscopic evaluation revealed serious problems with the circular stapler method. The cause of anastomotic stenosis may include the fact that if anastomosis is performed by a circular stapler method, all the layers of gastrointestinal tract are punched out at the anastomosis portion, and mucosal conjugation is not observed and the muscular layer is exposed in the inner lumen of the gastrointestinal tract. Taking that the ulcer is formed circularly at the anastomotic portion into account, it is easily understood that this circular ulcer develops into stenosis in the healing process. The advantage of triangular anastomosis for esophago-gastric anastomosis is less suture failure, and is extremely advantageous for prevention of stenosis at the anastomotic portion when compared with other anastomosis methods. However, with regard to the healing process of eversion anastomosis in gastrointestinal tract instrumental anastomosis, detailed examination is expected hereafter. PMID- 15900238 TI - Surgical treatment for lung cancer patients with poor pulmonary function. AB - This retrospective study was aimed to review the risk factors of postoperative hospital death in lung cancer patients with poor pulmonary reserves. We performed surgery on 30 lung cancer patients (average age: 71 years) with less than 1.0L of preoperative forced expiratory volume in one second (FEV1.0 < 1.0L) between 1982 and 2003. The preoperative FEV1.0 of these 30 patients was 0.81+/-0.1L on average. Six of 20 patients who underwent surgery between 1982 and 1997 died during their postoperative hospital stay (hospital death group). Between 1998 and 2003, 10 patients underwent surgery with uneventful postoperative courses. The clinical background was evaluated between the hospital death group (n = 6) and the survivor group (n = 24). The single-variable analysis of the preoperative conditions of the postoperative hospital death revealed the presence of two or more preoperative concomitant diseases (p < 0.001) and a performance status of grade 2 or higher (p = 0.005). Peripheral obstructive pneumonia with abscess and chronic pleurisy with adhesion influenced surgery and related to their postoperative hospital death. Whereas, patients with chronic obstructive pulmonary diseases (COPD) tended to show that pulmonary resection resulted in an improvement of pulmonary functions. CONCLUSION: To achieve better results of surgical treatment for patients with preoperative FEV1.0 < 1.0L, patient selection should be required based on precise evaluation of physical conditions and management of infectious diseases. Moreover, we thought that a preoperative performance status of grade 1 or lower, at most one preoperative concomitant disease, and a COPD are desirable for deciding the surgical indication. PMID- 15900239 TI - Conscious off pump coronary artery bypass surgery--an audit of our first 151 cases. AB - PURPOSE: An audit of our first 151 cases of conscious off pump coronary artery bypass (COPCAB) surgery with epidural anesthesia as sole anesthetic. METHODS: Patients underwent conscious off pump coronary artery bypass (OPCAB) surgery using high thoracic epidural anesthesia. The epidural catheter was inserted on the day before the surgery. RESULTS: There were 118 male and 33 female patients. The incision was via midsternotomy except in 3 patients. Single graft was performed in 25 patients, double in 61, triple in 46, quadruple in 19. Twenty nine patients developed pneumothorax. Three patients required conversion to general anesthesia. In one patient cardiopulmonary bypass (CPB) was instituted. There was no mortality in the group. CONCLUSION: Our experience shows that conscious OPCAB surgery can be performed safely in selected patients. PMID- 15900240 TI - Early results of aortic connector system for proximal anastomosis of the saphenous vein graft. AB - PURPOSE: Our objective is to clarify the technical points in using the symmetry aortic connector system (ACS) compared with hand-sewing from clinical and angiographic results of the patients who underwent off-pump coronary artery bypass grafting (CABG). METHODS: Among 84 patients who underwent CABG from January 2002 to August 2003, 36 off-pump CABGs were performed using at least one saphenous vein graft (SVG). Thirty proximal anastomosis of 17 patients were performed by ACS, and 30 proximal anastomosis of 19 patients were performed by hand-sewing under aortic side-clamping. Angiogram of the coronary arteries and grafts was carried out 18 days after the operation. RESULTS: Comparing the preoperative characteristics and intraoperative characteristics between the ACS group and hand-sewing group, revealed no significant factors at all. Postoperative angiography of coronary arteries and grafts revealed that the four SVGs with ACS were occluded probably because of kinking near the proximal anastomosis site. On the other hand, all SVGs anastomosed by hand-sewing were patent. CONCLUSION: ACS is very useful with easy handling during off-pump CABG, but it has potential limitations. In our limited experience, we have to mind the graft length as well as the position of proximal anastomosis so that the graft does not kink. PMID- 15900241 TI - Postoperative bronchial stump fistula responding well to occlusion with metallic coils and fibrin glue via a tracheostomy: a case report. AB - An 80-year-old man underwent middle and lower lobectomy of the right lung to treat squamous cell carcinoma (SCC) (4 cm in diameter) originating from the right B4 bronchus. On the 4th postoperative day, a massive air leak from the thoracic drain was noted. At that time, a diagnosis of bronchial stump fistula was made on the basis of the bronchoscopic findings. Continuous thoracic drainage, aspiration of sputum via a tracheostomy and intravenous administration of antibiotics were performed immediately after the diagnosis. However, the patient's condition was complicated by aspiration pneumonia. On the 11th postoperative day, bronchoscopic procedure to close the bronchial fistula was performed via the tracheostomy. During this procedure, metallic coils were first inserted into the fistula to serve as the core for occlusion. Then, instead of directly infusing fibrin glue, several small beans-sized pieces of Surgicell cotton (Johnson & Johnson Co., Cincinnati, OH) soaked in fluid A (concentrated fibrinogen) and the same number of Surgicell cotton pieces soaked in fluid B (thrombin) were alternately inserted into the fistula, to allow closure of the bronchial fistula. After this procedure, the embolus created remained in place without being expectorated, and the aspiration pneumonia entered remission, allowing the patient to be discharged from the hospital on the 24th postoperative day. At preset, 14 months after surgery, the patient is in good condition. This technique allows simple and reliable closure of a fistula if a tracheostomy is available. It should be selected as a therapy of first choice when dealing with patients with a postoperative bronchial stump fistula in poor general condition. Patients undergoing right pneumonectomy or middle and lower lobectomy of the right lung should be considered as belonging to a high risk group for bronchial fistula and as requiring preventable measures (e.g., covering the stump with an intercostal muscle flap). PMID- 15900242 TI - Primary lung cancer arising from the wall of a giant bulla. AB - We report a 58-year-old man who underwent surgical treatment of primary lung cancer arising from the wall of a giant bulla. Chest roentgenography and computed tomography revealed multiple emphysematous bullae in the bilateral upper lobes, and a right upper giant bulla with a mass measuring 6 cm arising on the bulla wall. Right upper lobectomy was performed, the postoperative pathological diagnosis was large cell carcinoma arising from the wall of a giant bulla. Although the postoperative course was uneventful and he was discharged, he underwent partial resection of the jejunum for recurrence of carcinoma in the jejunum, and postoperative chemotherapy, and he was alive 20 months after that operation. In general, patients with both pulmonary bullous disease and primary lung cancer have a very poor prognosis, because they receive treatment when the tumor is at an advanced stage. On the basis of our review of the literature, we recommend that middle-age male patients with a giant bulla who smoke should have annual chest roentgenography and/or chest computed tomography to screen for lung cancer arising in or close to the bullous disease, and that a giant bulla should be resected in patients older than 50 years because of the high incidence of coexisting cancer and bulla, to improve the prognosis of this disease. PMID- 15900243 TI - A case of invasive thymoma with endobronchial growth. AB - We report a rare case of invasive thymoma with endobronchial growth in a 69- year old woman. Chest CT revealed an ill-defined mass with a calcified lesion in the anterior mediastinum and coin lesion in the bilateral lung fields. An endobronchial polypoid tumor in the right B3 bronchus was found by bronchoscopic examination. A biopsy specimen obtained from the polypoid tumor suggested thymoma. After radiotherapy combined with chemotherapy, thymothymectomy combined with right upper lobectomy through median sternotomy was performed. The surgical specimen revealed that the tumor extended nodularly, not only into the pulmonary parenchyma but also into the bronchial lumen. We performed an added chemotherapy for lung metastasis, and partial resection of lung with video-assisted thoracoscopic surgery (VATS) was done. Now she is alive and disease-free. PMID- 15900244 TI - Video-assisted thoracic surgery for Ewing's sarcoma of the mediastinum in a 3 year-old girl. AB - We report of the experience of video-assisted thoracic surgery (VATS) for a Ewing's sarcoma at the posterior mediastinum in a 3 year old girl. The patient developed common cold like symptoms and developed rapid lower limb paraplegia. A chest computed tomography (CT) showed a solid mass adjacent to the vertebrae. This was diagnosed as a Ewing's sarcoma histopathologically by CT-guided needle biopsy (CT-NB). The mass was pressing on a nerve root in the vicinity of a vertebral foramen. As the tumor reduced after induction chemotherapy, with improvement of paraplegia, the patient underwent thoracoscopic examination followed by complete removal of the tumor by VATS. After surgery, the patient underwent postoperative chemo-radiotherapy. Five years has passed after operation without tumor recurrence. VATS could be an option as combined therapy for Ewing's sarcoma when well controlled and localized under induction therapy. PMID- 15900245 TI - A surgical case of ventricular septal perforation after repairing left ventricular free wall rupture. AB - A 78-year-old woman with diagnosis of acute myocardial infarction (AMI) in the anteroseptal area fell into cardiogenic shock suddenly just before starting percutaneous coronary intervention (PCI). Echocardiography showed left ventricular free wall rupture, then an emergent operation was performed by sutureless patch repair using collagen fleece with fibrinogen-based impregnation. Eight days later from the initial operation, the onset of ventricular septal perforation (VSP) was recognized. Fifteen days after, the infarct exclusion technique with endocardial patch was performed. She has been doing well 4 months after the operation without residual shunt. To our best knowledge, this is the first surgical case report that free wall rupture of left ventricle and VSP which are serious complications after myocardial infarction happened in succession. PMID- 15900246 TI - Re-do mitral valve replacement using the valve-on-valve technique: a case report. AB - We report a repeated mitral valve replacement (re-do MVR) using the valve-on valve technique for a degenerated bioprosthesis. A 49-year-old female, who had had a 29 mm Carpentier-Edwards mitral bioprosthesis for mitral regurgitation 20 years previously, was referred to our institution for dyspnea. She presented with pulmonary edema secondary to severe mitral bioprosthetic valve regurgitation. We replaced the degenerated mitral bioprosthesis with a 25 mm mechanical prosthesis using the valve-on-valve technique, as the struts of the bioprosthesis were embedded in the left ventricular myocardium. Removal of the bioprosthesis may be not only time-consuming but also complicated by cardiac rupture at the atrioventricular junction or the posterior left ventricular wall. The valve-on valve technique is a simplified procedure that can avoid the potential complications of complete excision of the bioprosthesis. We believe this technique can be a useful strategy for patients with a degenerated mitral bioprosthesis. PMID- 15900247 TI - A surgical case of concomitant aneurysms of the brachiocephalic and coronary arteries. AB - Brachiocephalic artery aneurysm with concomitant coronary artery aneurysm is rare. We describe a case of a patient with a history of prosthetic graft placement following resection of an abdominal aortic aneurysm and was subsequently found to have a brachiocephalic artery aneurysm. After surgical correction of the brachiocephalic aneurysm, postoperative coronary arteriography demonstrated coronary artery aneurysms, and the patient subsequently underwent coronary artery bypass grafting (CABG). PMID- 15900248 TI - Aortic root remodeling for root aneurysm with a uni-commissural aortic valve: report of a case. AB - A 23-year-old man was admitted for an aortic root aneurysm with mild aortic valve regurgitation (AR) and a small pressure gradient. At surgery, findings of aortic valve, one normal left posterior commissure and very rudimentary right anterior commissure, was compatible with the uni-commisural aortic valve. Aortic root replacement with valve-sparing technique was performed. Four years later no residual AR was observed. PMID- 15900249 TI - Intimal sarcoma of the thoracic aorta; clinical-course and autopsy finding. AB - We report a primary malignant tumor of the thoracic aorta with clinical-course from onset to death. A 63-year-old male was admitted to our hospital with intermittent claudication and bilateral lower extremity pain. The diagnosis was established after an abdominal operation. The tumor was subsequently resected including the thoracic aorta and replaced with a dacron graft. The pathological finding was of a primary intimal sarcoma. The patient lived for more than two years excluding the hospitalized period after the diagnosis. PMID- 15900250 TI - Surgical treatment of methicillin-resistant Staphylococcus aureus infection following infrainguinal arterial reconstruction. AB - A 64-year-old man was referred to our hospital with Methicillin-resistant Staphylococcus aureus (MRSA) infection following infrainguinal arterial reconstruction. As repeated MRSA sepsis occurred, we decided to remove the infected graft with distal revascularization via circuitous graft tunneling to avoid serious infections and allow limb salvage. An iliofemoro bypass was performed via an extra-anatomical bypass, from just below the iliac crest into the musculus quadriceps femoris using an 8 mm-ringed polyester gelatin polypropylene tube graft, with complete debridement of a groin infection. Postoperative 3-dimentional CT angiography revealed that the prostheses was patent and the patient had an uneventful postoperative course. We concluded that this extra-anatomical bypass was a safe procedure and an excellent option for patients with an infected vascular prosthetic graft in the groin after previous revascularization, like in our case with no available autogeneous vein grafts. PMID- 15900257 TI - Thrombosis risk assessment as a guide to quality patient care. PMID- 15900258 TI - Bilateral lower extremity duplex scanning revisited. PMID- 15900259 TI - The diagnosis of acute venous thromboembolism. PMID- 15900260 TI - Patent foramen ovale: the missing link between deep venous thrombotic disease and embolic stroke. PMID- 15900261 TI - The treatment of deep venous thrombosis, including the newer agents. PMID- 15900262 TI - Why warfarin and heparin need to overlap when treating acute venous thromboembolism. PMID- 15900263 TI - Management of pulmonary embolism in 2005. PMID- 15900264 TI - The role of fibrinolytic therapy in the treatment of venous thromboembolism. PMID- 15900265 TI - Immediate ambulation and leg compression in the treatment of deep vein thrombosis. PMID- 15900266 TI - Heparin-induced thrombocytopenia. PMID- 15900267 TI - Deep vein thrombosis and cancer: survival, recurrence, and anticoagulant choices. PMID- 15900268 TI - Long-term anticoagulation prophylaxis following acute thromboembolism. PMID- 15900269 TI - Practical aspects of the postthrombotic syndrome. PMID- 15900270 TI - CEAP classification for chronic venous disease. PMID- 15900271 TI - Perioperative bridging therapy for the at-risk patient on chronic anticoagulation. PMID- 15900272 TI - What's new for DVT prophylaxis for the medically ill. PMID- 15900273 TI - Air travel-related venous thromboembolism. PMID- 15900274 TI - Oral anticoagulants--the old and the new. PMID- 15900280 TI - Cytochrome P450 2C8: substrates, inhibitors, pharmacogenetics, and clinical relevance. AB - Cytochrome P450 (CYP) 2C8 [corrected] has been a relatively neglected member of the human CYP2C family. Over the period from 2000 through 2003, PubMed searches with the key word CYP2C8 returned only 10% to 15% of the citations obtained for all of the CYP2C enzymes combined. However, in the past year a crystal structure for CYP2C8 has been described, new inhibitors and probe substrates for the enzyme have been in development, the first case study was published linking CYP2C8 genetic polymorphisms to a disease state, and there has been an increasing awareness of the role that CYP2C8 plays in the disposition of therapeutic agents, especially from the pharmacogenetic and drug-drug interaction perspectives. This report discusses baseline characteristics of the enzyme and summarizes recent developments in these areas and their clinical relevance. PMID- 15900281 TI - CYP2C9 haplotype structure in European American warfarin patients and association with clinical outcomes. AB - OBJECTIVE: The goal of this study was to define the haplotype structure of the cytochrome P450 (CYP) 2C9 gene in a European American population and evaluate associations between CYP2C9 haplotypes and anticoagulation-related outcomes. METHODS: Genomic deoxyribonucleic acid from 192 European American patients stabilized on warfarin therapy was resequenced across 60 kilobases of the CYP2C9 genomic region, including all exons, dense sampling of introns, approximately 10 kilobases of the 5'-flanking region, and approximately 1.7 kilobases of the 3' untranslated region. RESULTS: A total of 132 single nucleotide polymorphisms (SNPs) were detected, of which 47 were present in the 5'-flanking promoter region, 11 in the exonic coding region, and 74 in the intron regions. Nine nonsynonymous SNPs in the coding region consisted of CYP2C9*2 , *3 , *9 , *11 , and *12 ; R125H; and 3 new structural variants. Sixty SNPs were present at a minor allele frequency of greater than 5%, and from this subpopulation, 23 haplotypes were inferred. Clustering analysis identified 6 major groups of related haplotypes that were further designated 1A, 1B, 1C, 1D, 2, or 3. The *2 and *3 SNPs appeared exclusively in groups 2 and 3, and these groups combined were associated with significantly reduced warfarin maintenance doses, longer time to stable dosing, and increased risk of bleeding. In contrast, combinations of haplotypes 1A, 1B, 1C, and 1D were not associated with differences in any of these outcomes. CONCLUSION: These data establish a whole-gene, high-resolution haplotype structure for CYP2C9 in a European American patient population and suggest that genetic variation in exons, rather than the promoter or other regulatory regions, is largely responsible for warfarin sensitivity associated with CYP2C9 variants in this population. PMID- 15900282 TI - Common genetic variants of microsomal epoxide hydrolase affect warfarin dose requirements beyond the effect of cytochrome P450 2C9. AB - BACKGROUND: Warfarin dose response is partially explained by the polymorphisms in the cytochrome P450 (CYP) 2C9 gene, affecting S -warfarin clearance, as well as by age and body weight. We examined the influence on warfarin dose requirements of candidate genes encoding microsomal epoxide hydrolase (mEH), as well as glutathione S -transferase A1 (GSTA1) components of vitamin K epoxide reductase and the gamma-glutamylcarboxylase (GGCX) gene. METHODS: We studied the effects of CYP2C9, mEH, GSTA1, and GGCX genotypes on warfarin maintenance doses, accounting for age, weight, vitamin K plasma concentrations and concurrent medications, in 100 patients undergoing therapeutic anticoagulation. RESULTS: Allele frequencies were 76.5%, 12.5%, and 11% for CYP2C9*1 , *2 , and *3 , respectively; 75% and 25% for mEH T 612 C; 75.8% and 24.2% for mEH A 691 G; 73.5% and 26.5% for GSTA1 T 631 G; and 70.5% and 29.5% for GGCX G 8762 A. Warfarin doses differed among the CYP2C9 ( 2C9*1 , 2C9*2 , and 2C9*3 ) genotype groups: 6.3 +/- 1.9 mg/d, 5.3 +/- 1.8 mg/d, and 3.8 +/- 1.7 mg/d, respectively (F = 4.83, P < .01). There were no differences in any of the other genotype groups. Among the 62 wild-type CYP2C9 patients, variant mEH T 612 C homozygotes required higher doses than heterozygotes and wild-type patients (7.5 +/- 2.9 mg/d, 6.5 +/- 4.2 mg/d, and 6.0 +/- 2.6 mg/d, respectively [F = 3.57, P = .03]). The odds ratio for requiring greater than 7 mg/d in variant mEH T 612 C patients versus wild-type patients was 3.14 (95% confidence interval, 1.47-6.67), accounting for CYP2C9. CONCLUSIONS: Variant mEH T 612 C genotypes are associated with warfarin doses of greater than 50 mg/wk beyond the effect of CYP2C9. PMID- 15900284 TI - Genotype-phenotype associations of cytochrome P450 3A4 and 3A5 polymorphism with midazolam clearance in vivo. AB - The molecular basis for the wide interindividual variability of cytochrome P450 (CYP) 3A metabolic activity was studied in vivo at a genetic level. A single oral dose of midazolam was administered to 26 healthy subjects. The variability in midazolam oral clearance was 11-fold. No differences in midazolam oral clearance related to gender or ethnicity were observed. Selective sequencing of CYP3A4 and CYP3A5 genes revealed 18 single nucleotide polymorphisms (SNPs), including 8 novel CYP3A4 SNPs. Thirteen novel CYP3A4 haplotypes, 2 novel CYP3A5 haplotypes, and 1 major novel multigene haplotype ( CYP3A4*VI - CYP3A5*3A ) were also identified. No significant genotype-phenotype or haplotype-phenotype associations were found for any of the SNPs or haplotypes studied, including CYP3A4*1B , CYP3A5*3 , and CYP3A5*6 , even when ethnicity was considered. The only exceptions were the haplotype CYP3A4*VI and the multigene haplotype CYP3A4*VI - CYP3A5*3A . The carriers of the haplotype CYP3A4*VI had a 1.8-fold higher clearance of midazolam in black subjects (ANOVA on ranks, P = .028) compared with other individuals, and the carriers of the multigene haplotype CYP3A4*VI - CYP3A5*3A had a 1.7-fold higher clearance in the entire population (ANOVA on ranks, P = .012). In conclusion, these results indicate that the genetic variants identified so far in the CYP3A4 and CYP3A5 genes have only a limited impact on CYP3A mediated drug metabolism in vivo. PMID- 15900285 TI - Human response to alpha2-adrenergic agonist stimulation studied in an isolated vascular bed in vivo: Biphasic influence of dose, age, gender, and receptor genotype. AB - BACKGROUND AND OBJECTIVES: Activation of alpha 2 -adrenergic receptors regulates a broad spectrum of physiologic responses, including blood pressure (centrally and peripherally), sedation, analgesia, insulin release, renal function, cognition, and behavior. The purpose of this study was to explore systematically the local vascular responses in humans triggered by a highly selective alpha 2 adrenergic agonist (azepexole [B-HT 933]) and whether such responses are dose dependent or influenced by age, gender, or allelic variation at the drug's receptor. METHODS: We evaluated dorsal hand vein vascular responses to the infusion of a wide spectrum of doses of azepexole, assessing any venodilation, as well as the maximal extent of venoconstriction (B max ) and the dose that produced a half-maximal effect (K d ), in 50 healthy normotensive adults of both genders and 4 ethnicities. Genomic deoxyribonucleic acid from the study subjects was evaluated at polymorphisms of the alpha 2B -adrenergic receptor gene (ADRA2B). RESULTS: We found previously unreported initial venodilation to low doses (10-100 ng/min) of azepexole, followed by progressive, intense venoconstriction to higher doses (200-100,000 ng/min) of the drug. Younger individuals (aged <30 years) had less venodilation than older individuals (aged >30 years) with low doses of azepexole but had a greater extent of venoconstriction at higher doses of azepexole (ANOVA, P = .001). Men had less venodilation than women with low doses of azepexole but greater venoconstriction with higher doses (ANOVA, P = .036). Several common polymorphisms (>10% minor allele frequency) at ADRA2B (insertion/deletion polymorphism [Glu 322-325 ], G 98C, C1182A, and C1776A) did not show an association with either B max or K d for the drug response. The A36G (Thr12Thr) synonymous single nucleotide polymorphism displayed a nonsignificant trend (P = .073) toward higher K d in A/G heterozygotes compared with A/A homozygotes. CONCLUSIONS: Local infusion into the human dorsal hand vein of a highly selective alpha 2 -adrenergic agonist, azepexole, produces biphasic responses, with venodilation at a low dose and intense venoconstriction at a higher concentration. These responses to azepexole show prominent differences as a function of age and gender but appear not to depend on common allelic variations at the ADRA2B receptor. PMID- 15900286 TI - Effects of gemfibrozil, itraconazole, and their combination on the pharmacokinetics of pioglitazone. AB - BACKGROUND AND OBJECTIVE: The thiazolidinedione antidiabetic drug pioglitazone is metabolized mainly by cytochrome P450 (CYP) 2C8 and CYP3A4 in vitro. Our objective was to study the effects of gemfibrozil, itraconazole, and their combination on the pharmacokinetics of pioglitazone to determine the role of these enzymes in the fate of pioglitazone in humans. METHODS: In a randomized, double-blind, 4-phase crossover study, 12 healthy volunteers took either 600 mg gemfibrozil or 100 mg itraconazole (first dose, 200 mg), both gemfibrozil and itraconazole, or placebo twice daily for 4 days. On day 3, they received a single dose of 15 mg pioglitazone. Plasma drug concentrations and the cumulative excretion of pioglitazone and its metabolites into urine were measured for up to 48 hours. RESULTS: Gemfibrozil alone raised the mean total area under the plasma concentration-time curve from time 0 to infinity [AUC(0-infinity)] of pioglitazone 3.2-fold (range, 2.3-fold to 6.5-fold; P < .001) and prolonged its elimination half-life (t (1/2) ) from 8.3 to 22.7 hours ( P < .001) but had no significant effect on its peak concentration (C max ) compared with placebo (control). Gemfibrozil increased the 48-hour excretion of pioglitazone into urine by 2.5-fold ( P < .001) and reduced the ratios of the active metabolites M-III and M-IV to pioglitazone in plasma and urine. Gemfibrozil decreased the area under the plasma concentration-time curve from time 0 to 48 hours [AUC(0-48)] of the metabolites M-III and M-IV by 42% ( P < .05) and 45% ( P < .001), respectively, but their total AUC(0-infinity) values were reduced by less or not at all. Itraconazole had no significant effect on the pharmacokinetics of pioglitazone and did not alter the effect of gemfibrozil on pioglitazone pharmacokinetics. The mean area under the concentration versus time curve to 49 hours [AUC(0-49)] of itraconazole was 46% lower ( P < .001) during the gemfibrozil-itraconazole phase than during the itraconazole phase. CONCLUSIONS: Gemfibrozil elevates the plasma concentrations of pioglitazone, probably by inhibition of its CYP2C8-mediated metabolism. CYP2C8 appears to be of major importance and CYP3A4 of minor importance in pioglitazone metabolism in vivo in humans. Concomitant use of gemfibrozil with pioglitazone may increase the effects and risk of dose-related adverse effects of pioglitazone. However, studies in diabetic patients are needed to determine the clinical significance of the gemfibrozil-pioglitazone interaction. PMID- 15900288 TI - A pharmacokinetic-pharmacodynamic model for the mobilization of CD34+ hematopoietic progenitor cells by AMD3100. AB - BACKGROUND: AMD3100 is a small-molecule CXCR4 antagonist that has been shown to induce the mobilization of CD34 + hematopoietic progenitor cells from bone marrow to peripheral blood. AMD3100 has also been shown to augment the mobilization of CD34 + cells in cancer patients when administered in combination with granulocyte colony-stimulating factor (G-CSF) (filgrastim). The purpose of this study was to characterize the exposure-response relationship of AMD3100 in mobilizing CD34 + cells when administered as a single agent in healthy volunteers. METHODS: AMD3100 concentrations and CD34 + cell counts obtained from 29 healthy subjects in a single-dose, intensively sampled pharmacokinetic/pharmacodynamic (PK-PD) study were analyzed by use of nonlinear mixed effects regression with the software NONMEM. FOCE (first order conditional estimation) with interaction was the estimation method, and simultaneous PK-PD fitting was adopted. RESULTS: The pharmacokinetics of AMD3100 was described by a 2-compartment model with first order absorption. The population estimates (+/-SE) for clearance and central volume of distribution were 5.17 +/- 0.49 L/h and 16.9 +/- 3.79 L, respectively. CD34 + cell mobilization was best described by an indirect effect model that stimulates the entry process of CD34 + from bone marrow to peripheral blood in the form of a sigmoid maximum effect model. The population estimates (+/-SE) of maximum effect, concentration causing 50% of maximum response, and equilibration time were 12.6 +/- 4.89, 53.6 +/- 11.9 mug/L, and 5.37 +/- 1.31 hours, respectively. CONCLUSIONS: This study characterizes the exposure-response relationship of AMD3100 in mobilizing CD34 + cells after subcutaneous administration. This PK-PD model will be useful in assessing relevant covariates and for optimizing the use of AMD3100 in various patient populations. PMID- 15900287 TI - In vivo effects of goldenseal, kava kava, black cohosh, and valerian on human cytochrome P450 1A2, 2D6, 2E1, and 3A4/5 phenotypes. AB - OBJECTIVES: Phytochemical-mediated modulation of cytochrome P450 (CYP) activity may underlie many herb-drug interactions. Single-time point phenotypic metabolic ratios were used to determine whether long-term supplementation of goldenseal ( Hydrastis canadensis ), black cohosh ( Cimicifuga racemosa ), kava kava ( Piper methysticum ), or valerian ( Valeriana officinalis ) extracts affected CYP1A2, CYP2D6, CYP2E1, or CYP3A4/5 activity. METHODS: Twelve healthy volunteers (6 women) were randomly assigned to receive goldenseal, black cohosh, kava kava, or valerian for 28 days. For each subject, a 30-day washout period was interposed between each supplementation phase. Probe drug cocktails of midazolam and caffeine, followed 24 hours later by chlorzoxazone and debrisoquin (INN, debrisoquine), were administered before (baseline) and at the end of supplementation. Presupplementation and postsupplementation phenotypic trait measurements were determined for CYP3A4/5, CYP1A2, CYP2E1, and CYP2D6 by use of 1 hydroxymidazolam/midazolam serum ratios (1-hour sample), paraxanthine/caffeine serum ratios (6-hour sample), 6-hydroxychlorzoxazone/chlorzoxazone serum ratios (2-hour sample), and debrisoquin urinary recovery ratios (8-hour collection), respectively. The content of purported "active" phytochemicals was determined for each supplement. RESULTS: Comparisons of presupplementation and postsupplementation phenotypic ratio means revealed significant inhibition (approximately 40%) of CYP2D6 (difference, -0.228; 95% confidence interval [CI], 0.268 to -0.188) and CYP3A4/5 (difference, -1.501; 95% CI, -1.840 to -1.163) activity for goldenseal. Kava produced significant reductions (approximately 40%) in CYP2E1 only (difference, -0.192; 95% CI, -0.325 to -0.060). Black cohosh also exhibited statistically significant inhibition of CYP2D6 (difference, -0.046; 95% CI, -0.085 to -0.007), but the magnitude of the effect (approximately 7%) did not appear to be clinically relevant. No significant changes in phenotypic ratios were observed for valerian. CONCLUSIONS: Botanical supplements containing goldenseal strongly inhibited CYP2D6 and CYP3A4/5 activity in vivo, whereas kava inhibited CYP2E1 and black cohosh weakly inhibited CYP2D6. Accordingly, serious adverse interactions may result from the concomitant ingestion of goldenseal supplements and drugs that are CYP2D6 and CYP3A4/5 substrates. Kava kava and black cohosh may interact with CYP2E1 and CYP2D6 substrates, respectively. Valerian appears to be less likely to produce CYP-mediated herb-drug interactions. PMID- 15900289 TI - Renal effects of the cyclooxygenase-inhibiting nitric oxide donator AZD3582 compared with rofecoxib and naproxen during normal and low sodium intake. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit cyclooxygenase (COX) and can thereby reduce renal function, especially with respect to solute excretion and renal perfusion. AZD3582 [4-(nitrooxy)butyl-(2S)-2-(6-methoxy-2 naphthyl)propanoate] is a COX-inhibiting nitric oxide donator. Donation of nitric oxide by AZD3582 could preserve blood flow and thereby counteract the deleterious effects of COX inhibition in the gastrointestinal tract and possibly in other organ systems, including the kidney. The aim of this single-dose study was to assess the hypothesis that AZD3582 would not adversely affect renal function compared with NSAIDs. METHODS: In a parallel, randomized, double-blind fashion, a total of 60 healthy subjects (age range, 20-44 years) received 2 single doses of 750 mg AZD3582, 1500 mg AZD3582, 50 mg rofecoxib, 500 mg naproxen, or placebo (n = 12 per group). The first dose was given after a 5-day normal-sodium diet (150 mmol/d), and the second was given after a consecutive 3-day low-sodium diet (10 mmol/d). Urinary sodium excretion during normal sodium intake and glomerular filtration rate (GFR) (assessed by iohexol clearance) during sodium depletion were the primary variables measured. RESULTS: Urinary sodium excretion was reduced in all active treatment groups (maximal reduction of approximately 11 mmol/h during normal sodium intake, P < .05 versus placebo for all groups). GFR was also reduced in all active treatment groups. In sodium-depleted subjects, the mean (SD) maximal reduction in GFR during 0 to 6 hours for 750 mg AZD3582, 1500 mg AZD3582, 50 mg rofecoxib, and 500 mg naproxen was 28.1 mL/min (13.5 mL/min), 33.7 mL/min (23.3 mL/min), 25.2 mL/min (29.2 mL/min), and 41.7 mL/min (30.7 mL/min), respectively, with a statistically significant difference between 500 mg naproxen and placebo. Relative changes in sodium excretion and GFR were similar during normal sodium intake and sodium depletion during active treatment. CONCLUSION: The renal effects of 750 mg and 1500 mg AZD3582 were similar to those of 500 mg naproxen and 50 mg rofecoxib. Thus the potential for nitric oxide donation to create a renal-sparing agent was not found for AZD3582. PMID- 15900290 TI - Linezolid and rifampin: Drug interaction contrary to expectations? PMID- 15900291 TI - Herbal medicines in Malaysia. PMID- 15900292 TI - Traditional aqueous kava extracts inhibit cytochrome P450 1A2 in humans: Protective effect against environmental carcinogens? PMID- 15900295 TI - Hereditary colorectal cancer-part II. PMID- 15900296 TI - Sociodemographic factors and delays in the diagnosis of six cancers: analysis of data from the "National Survey of NHS Patients: Cancer". AB - This paper aims to explore the relationship between sociodemographic factors and the components of diagnostic delay (total, patient and primary care, referral, secondary care) for these six cancers (breast, colorectal, lung, ovarian, prostate, or non-Hodgkin's lymphoma). Secondary analysis of patient-reported data from the 'National Survey of NHS patients: Cancer' was undertaken (65 192 patients). Data were analysed using univariate analysis and Generalised Linear Modelling. With regard to total delay, the findings from the GLM showed that for colorectal cancer, the significant factors were marital status and age, for lung and ovarian cancer none of the factors were significant, for prostate cancer the only significant factor was social class, for non-Hodgkin's lymphoma the only significant factor was age, and for breast cancer the significant factors were marital status and ethnic group. Where associations between any of the component delays were found, the direction of the association was always in the same direction (female subjects had longer delays than male subjects, younger people had longer delays than older people, single and separated/divorced people had longer delays than married people, lower social class groups had longer delays than higher social class groups, and Black and south Asian people had longer delays than white people). These findings should influence the design of interventions aimed at reducing diagnostic delays with the aim of improving morbidity, mortality, and psychological outcomes through earlier stage diagnosis. PMID- 15900297 TI - An overview of menopausal oestrogen-progestin hormone therapy and breast cancer risk. AB - Results from the Women's Health Initiative (WHI) trial support findings from observational studies that oestrogen-progestin therapy (EPT) use is associated with an increase in breast cancer risk. We conducted a meta-analysis using EPT specific results from the Collaborative Group on Hormonal Factors in Breast Cancer (CGHFBC) pooled analysis and studies published since that report to obtain an overview of EPT use and breast cancer risk. We also assessed risk by histologic subtype of breast cancer, by schedule of the progestin component of EPT, and by recency of use. We estimate that overall, EPT results in a 7.6% increase in breast cancer risk per year of use. The risk was statistically significantly lower in US studies than in European studies - 5.2 vs 7.9%. There was a significantly higher risk for continuous-combined than for sequential EPT use in Scandinavian studies where much higher total doses of progestin were used in continuous-combined than in sequential EPT. We observed no overall difference in risk for lobular vs ductal carcinoma but did observe a slightly higher risk for current vs past EPT use. PMID- 15900298 TI - Stage 4 neuroblastoma: sequential hemi-body irradiation or high-dose chemotherapy plus autologous haemopoietic stem cell transplantation to consolidate primary treatment. AB - The aim of the present study was to evaluate the effectiveness of two consecutive nonrandomised treatment programs applied between 1989 and 1999 at the Istituto Nazionale Tumori of Milan in an unselected cohort of 59 children over the age of one with stage 4 neuroblastoma. Both treatment programs consisted of two phases, the induction of the remission phase and the consolidation phase. The induction of the remission phase consisted of intensive chemotherapy, and remained the same throughout the study period. The consolidation phase consisted of sequential hemi body irradiation (HBI) (10 Gy per session, 6 weeks apart) in the first period (1988-June 1994) and sequential high-dose cyclophosphamide, etoposide, mitoxantrone+L-PAM and autologous haemopoietic stem cell transplantation in the second (July 1994-1999). Intention-to-treat analysis revealed a significantly better outcome for patients treated with the second program, the 5-year event free survival probability being 0.12 for program 1 and 0.31 for program 2 (P=0.03). This finding led us to conclude that sequential HBI is useless as consolidation treatment. The high-dose chemotherapy adopted in the second program enabled a proportion of patients to obtain long-term survival but, since the clinical results remain unsatisfactory, new treatment strategies are warranted. PMID- 15900299 TI - The metastasis associated protein S100A4: role in tumour progression and metastasis. AB - The metastasis associated protein S100A4 is a small calcium binding protein that is associated with metastatic tumors and appears to be a molecular marker for clinical prognosis. Below we discuss its biochemical properties and possible cellular functions in metastasis including cell motility, invasion, apoptosis, angiogenesis and differentiation. PMID- 15900300 TI - Analysis of differential gene expression in human melanocytic tumour lesions by custom made oligonucleotide arrays. AB - Melanoma is one of the most aggressive types of cancer and resection of the tumour prior to dissemination of tumour cells is still the most effective treatment. Therefore, early diagnosis of melanocytic lesions is important and identification of novel (molecular) markers would be helpful to improve diagnosis. Moreover, better understanding of molecular targets involved in melanocytic tumorigenesis could possibly lead to development of novel interventions. In this study, we used a custom made oligonucleotide array containing 298 genes that were previously found to be differentially expressed in human melanoma cell lines 1F6 (rarely metastasising) and Mel57 (frequently metastasising). We determined differential gene expression in human common nevocellular nevus and melanoma metastasis lesions. By performing nine dye-swap array experiments, using individual as well as pooled melanocytic lesions, a constant differential expression could be detected for 25 genes in eight out of nine or nine out of nine array analyses. For at least nine of these genes, namely THBD, FABP7, H2AFJ, RRAGD, MYADM, HR, CKS2, NCK2 and GDF15, the differential expression found by array analyses could be verified by semiquantitative and/or real-time quantitative RT-PCR. The genes that we identified to be differentially expressed during melanoma progression could be potent targets for diagnostic, prognostic and/or therapeutic interventions. PMID- 15900301 TI - Personality and cancer survival: the Miyagi cohort study. AB - We tested the hypothesis that personality plays a role in cancer outcome in a population-based prospective cohort study in Japan. In July 1990, 41 442 residents of Japan completed a short form of the Eysenck Personality Questionnaire-Revised and a questionnaire on various health habits, and between January 1993 and December 1997, 890 incident cases of cancer were identified among them. These 890 cases were followed up until March 2001, and a total of 356 deaths from all causes was identified among them. Cox proportional-hazards regression was used to estimate the hazard ratio (HR) of death according to four score levels on each of four personality subscales (extraversion, neuroticism, psychoticism, and lie), with adjustment for potential confounding factors. Multivariable HRs of deaths from all causes for individuals in the highest score level on each personality subscale compared with those at the lowest level were 1.0 for extraversion (95% CI=0.8-1.4; Trend P=0.73), 1.1 for neuroticism (0.8 1.6; Trend P=0.24), 1.2 for psychoticism (0.9-1.6; Trend P=0.29), and 1.0 for lie (0.7-1.5; Trend P=0.90). The data obtained in this population-based prospective cohort study in Japan do not support the hypothesis that personality is associated with cancer survival. PMID- 15900302 TI - Reduced expression of chemokine (C-C motif) ligand-2 (CCL2) in ovarian adenocarcinoma. AB - Chemokine (C-C motif) ligand-2 (CCL2) is a chemoattractant and activator of macrophages and is a key determinant of the macrophage infiltrate into tumours. We demonstrate here that CCL2 is expressed in normal human ovarian surface epithelium (HOSE) cells and is silenced in most ovarian cancer cell lines, and silenced or downregulated in the majority of primary ovarian adenocarcinomas. Analysis of the CCL2 locus at 17q11.2-q12 showed loss of heterozygosity (LOH) in 70% of primary tumours, and this was significantly more common in tumours of advanced stage or grade. However, we did not detect any mutations in the CCL2 coding sequence in 94 primary ovarian adenocarcinomas. These data support the hypothesis that CCL2 may play a role in the pathobiology of ovarian cancers, but additional studies will be required to evaluate this possibility. PMID- 15900303 TI - Point mutation in GRIM-19: a new genetic lesion in Hurthle cell thyroid carcinomas. PMID- 15900306 TI - Direct comparison of dietary portfolio vs statin on C-reactive protein. AB - BACKGROUND: 3-Hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins) markedly reduce serum cholesterol and have anti-inflammatory effects. The effect of cholesterol-lowering diets on inflammatory biomarkers is less well known. OBJECTIVE: To compare the efficacy of a dietary combination (portfolio) of cholesterol-lowering foods vs a statin in reducing C-reactive protein (CRP) as a biomarker of inflammation linked to increased cardiovascular disease risk. METHODS: In all, 34 hyperlipidemic subjects completed three 1-month treatments as outpatients in random order: a very low-saturated fat diet (control); the same diet with 20 mg lovastatin (statin); and a diet high in plant sterols (1.0 g/1000 kcal), soy protein (21.4 g/1000 kcal), viscous fibers (9.8 g/1000 kcal), and almonds (14 g/1000 kcal) (portfolio). Fasting blood samples were obtained at weeks 0, 2, and 4. RESULTS: Using the complete data, no treatment reduced serum CRP. However, when subjects with CRP levels above the 75th percentile for previously reported studies (> 3.5 mg/l) were excluded, CRP was reduced similarly on both statin, -16.3 +/- 6.7% (n = 23, P = 0.013) and dietary portfolio, -23.8 +/- 6.9% (n = 25, P = 0.001) but not the control, 15.3 +/- 13.6% (n = 28, P = 0.907). The percentage CRP change from baseline on the portfolio treatment (n = 25) was greater than the control (n = 28, P = 0.004) but similar to statin treatment (n = 23, P = 0.349). Both statin and portfolio treatments were similar in reducing CRP and numerically more effective than control but only the change in portfolio was significant after the Bonferroni adjustment. CONCLUSIONS: A combination of cholesterol-lowering foods reduced C-reactive protein to a similar extent as the starting dose of a first-generation statin. PMID- 15900307 TI - Effects of soy supplementation on blood lipids and arterial function in hypercholesterolaemic subjects. AB - BACKGROUND: Studies on soy supplementation suggest a cardioprotective potential. OBJECTIVE: To examine the effects on LDL cholesterol and arterial function as a result of dietary enrichment with soy supplementation. DESIGN: A Randomized, double blind, parallel intervention trial. SETTING: Department of Endocrinology and Metabolism C, Aarhus University Hospital, and Department of Human Nutrition, The Royal Veterinary and Agricultural University, Denmark. SUBJECTS: In all, 100 hypercholesterolaemic but otherwise healthy subjects were included in the study of which 89 completed it. INTERVENTIONS: Subjects were randomly assigned to 24 weeks of daily intake of either a soy supplement, Abalon (30 g soy protein, 9 g cotyledon fibre and 100 mg isoflavones) or placebo (30 g of casein). The soy supplement and placebo were provided in two sachets daily that were stirred in water. Fasting plasma lipids, TNF-alpha, homocysteine, insulin sensitivity, homeostasis model assessment (HOMA-IR), serum insulin, serum glucose, blood pressure as well as Glucagon-like peptide 1 (GLP-1), glucose-dependent insulinotropic peptide (GIP) and plasma lipids to a fat-rich meal were recorded before and after the intervention. In a sub study in 32 subjects, arterial dilatory capacity, compliance, and distensibility were recorded before and after the intervention. RESULTS: In the main study, no difference in fasting plasma lipid levels or insulin sensitivity was found between soy-based supplement and placebo. A significant postprandial increase in GIP to the meal test was observed in the soy group (P < 0.05). In a substudy, no difference between the groups in changes in flow-mediated vasodilatation (P = 0.84) was detected, while the soy supplementation caused a reduction in LDL and total cholesterol. CONCLUSIONS: No significant effects on blood lipids were observed in the main study to a soy supplementation in hypercholesterolaemic subjects after 24 weeks. In the substudy, the soy supplementation, however, reduced LDL and total cholesterol but did not influence markers of arterial function. PMID- 15900308 TI - Lower resting metabolic rate and higher velocity of weight gain in a prospective study of stunted vs nonstunted girls living in the shantytowns of Sao Paulo, Brazil. AB - OBJECTIVE: Previous studies have shown that stunting increases the risk of obesity in developing countries, particularly among girls and women, but the underlying reasons are not known. This study aimed to investigate the relationship between stunting, weight gain, and resting metabolic rate. DESIGN AND SUBJECTS: A prospective study was conducted over 36 months with girls from shantytowns in Sao Paulo, Brazil. A total of 15 stunted girls (S) were compared with 15 nonstunted (N) ones of similar weight for height ratio. INTERVENTIONS: Resting metabolic rate was measured using indirect calorimetry, and the socioeconomic status was determined by interviews in the household. In addition, body composition was measured by skinfold thickness, while the growth rate was calculated dividing the change in weight and the change in height by the follow up period. RESULTS: The results of the present study, when combined, revealed that the S group had a lower resting metabolic rate throughout the follow-up period with the differences being significant at 24 and 36 months of follow-up, associated with an increase in the rate of weight gain and a decrease in lean mass, when compared to the N group. CONCLUSIONS: These changes are known to be risk factors for obesity and may help to explain the particularly higher prevalence of obesity in women in urban areas of developing countries. PMID- 15900309 TI - Iron intake and dietary sources of iron in Flemish adolescents. AB - OBJECTIVE: To investigate the dietary iron intake and food sources of iron in Flemish adolescents. DESIGN: Cross-sectional survey; dietary assessment method: a 7-day estimated food record. SETTING: Private and public secondary schools in Ghent, a city in the Dutch-speaking part of Belgium. SUBJECTS: A total of 341 adolescents (129 boys and 212 girls), 13-18 y, randomly selected by a multistage clustered sampling technique. RESULTS: The mean total iron intake (s.d.) for boys was 13.4 (+/- 2.91) mg/day and for girls 10.1 (+/- 2.79) mg/day. A proportion of 38.8% of the boys and 99.5% of the girls had a mean total iron intake below the Belgian Recommended Dietary Allowance and 3.1% of the boys and 71.2% of the girls below the British Estimated Average Requirement. When bioavailable iron intake is considered, 84.5% of the boys and only 16.5% of the girls met the age-specific requirement. The food groups with the highest mean proportional contribution to total iron intake in both males and females were bread, meat and meat products, cereals and potatoes. A comparison of adolescents from the highest tertile of iron intake (mg/day) with adolescents from the lowest tertile showed a significantly higher energy-adjusted intake of brown bread and a significantly lower intake of soft drinks in the former group in both boys and girls. A significantly higher energy-adjusted intake of breakfast cereals in adolescents of the highest tertile than those of the lowest tertile was seen in girls only. Analyses in consumers only did not change this overall picture. CONCLUSIONS: One can conclude that the mean iron intake of Flemish girls is considerably lower than the current recommendations. An increased iron intake in this subgroup of the population is therefore advisable. PMID- 15900310 TI - Correlates of regular fish consumption in French elderly community dwellers: data from the Three-City study. AB - OBJECTIVE: Several studies have found an inverse association between fish consumption and risk of dementia. The objective of this study was to describe the characteristics of older fish consumers, in order to identify potential confounders in this relationship. DESIGN: Cross-sectional analysis of the baseline data of a prospective cohort study. SETTING: Bordeaux, Dijon, and Montpellier (France) in 1999-2000. SUBJECTS: A total of 9280 community dwellers aged 65 y and above participating in the baseline examination of the Three-City Study. INTERVENTIONS: All participants answered a face-to-face interview and underwent a physical examination. Cross-sectional analyses were performed by logistic regressions. RESULTS: Regular fish consumers (at least weekly) had a higher education (OR from 1.19 to 1.65, P = 0.0003) and income (OR from 1.37 to 1.89, P < 0.0001). Controlling for age, sex, education, and city, they had a higher consumption of pulses, fruit, and vegetables (P < 0.001). They were more often alcohol drinkers (P < 0.0001). They felt in better health (P < 0.05), exhibited less depressive symptoms (P < 0.001), and scored higher on the Mini Mental Status Examination (P < 0.05). However, their objective physical health status was not better, except that they were leaner. They suffered more often from hypertension and past stroke. CONCLUSIONS: Socioeconomic status, dietary habits, depression, and vascular risk factors could act as confounders in the relationship between fish consumption and risk of dementia. PMID- 15900311 TI - Typing without calling the allele: a strategy for inferring SNP haplotypes. PMID- 15900312 TI - Farm Family Exposure Study: methods and recruitment practices for a biomonitoring study of pesticide exposure. AB - PURPOSE: The Farm Family Exposure Study was initiated to characterize pesticide exposure to farm family members around the time of one pesticide application in a manner that will facilitate exposure assessment in epidemiologic studies of pesticides. METHODS: A sample of farm families with children was recruited by randomly selecting farmers from lists of licensed pesticide applicators in Minnesota and South Carolina. Eligible families were selected from among those who planned to apply one of three chemicals, glyphosate, 2,4-D, or chlorpyrifos, as part of their normal operations. The applicator, spouse, and all children in the family ages 4-17 years were included in the study. The applicator and spouse completed self-administered questionnaires addressing demographics, farming practices and potential exposures to them and their children. Field observers documented the application, recorded application practices, equipment, potential exposures, and the presence of children or spouses in the immediate vicinity of pesticide activities. All study participants were asked to collect each urine void for 5 days, 1 day before through 3 days after the application. Pesticides were measured in 24-h composite urine samples with a one part per billion limit of detection. RESULTS: Of 11,164 applicators screened, 994 families met the inclusion criteria. Of these, 95 families were enrolled. Enrollees were similar in most characteristics to their peers who were not participants in the study. In total, there were 106 applications, 10 of which involved more than one chemical. This resulted in urinary data for 48 farmers and spouses and their 79 children for glyphosate, 34 farmers and spouses and their 50 children for chlorpyrifos, and 34 farmers and spouses and their 53 children for 2,4-D. Compliance with the 24-h urine collection was particularly good for the adult participants. There were more missing samples for children than for adults, but overall compliance was high. CONCLUSION: The Farm Family Exposure Study should provide insights about pesticide exposure under real world conditions and thereby facilitate improved exposure assessment in epidemiologic studies of agricultural populations. PMID- 15900314 TI - Initiation factor modifications in the preapoptotic phase. AB - Recent studies have identified several mechanistic links between the regulation of translation and the process of apoptosis. Rates of protein synthesis are controlled by a wide range of agents that induce cell death, and in many instances, the changes that occur to the translational machinery precede overt apoptosis and loss of cell viability. The two principal ways in which factors required for translational activity are modified prior to and during apoptosis involve (i) changes in protein phosphorylation and (ii) specific proteolytic cleavages. In this review, we summarise the principal targets for such regulation, with particular emphasis on polypeptide chain initiation factors eIF2 and eIF4G and the eIF4E-binding proteins. We indicate how the functions of these factors and of other proteins with which they interact may be altered as a result of activation of apoptosis and we discuss the potential significance of such changes for translational control and cell growth regulation. PMID- 15900315 TI - Internal ribosome entry segment-mediated translation during apoptosis: the role of IRES-trans-acting factors. AB - During apoptosis, there is a reduction in translation initiation caused by caspase cleavage of several of the factors required for the cap-dependent scanning mechanism. Under these circumstances, many proteins that are required for apoptosis are instead translated by the alternative method of internal ribosome entry. This mechanism requires the formation of a complex RNA structural element and in the presence of internal ribosome entry segment (IRES)-trans acting factors (ITAFs), the ribosome is recruited to the RNA. The interactions of several ITAFs with IRESs have been investigated in detail, and several mechanisms of action have been noted, including acting as chaperones, stabilising and remodelling the RNA structure. Structural remodelling by PTB in particular will be discussed, and how this protein is able to facilitate recruitment of the ribosome to several IRESs by causing previously occluded sites to become more accessible. PMID- 15900317 TI - Impaired object recognition memory following methamphetamine, but not p chloroamphetamine- or d-amphetamine-induced neurotoxicity. AB - Repeated moderate doses of methamphetamine (mAMPH) damage forebrain monoaminergic terminals and nonmonoaminergic cells in somatosensory cortex, and impair performance in a novelty preference task of object recognition (OR). This study aimed to determine whether the memory deficit seen after a neurotoxic mAMPH regimen results from damage to dopamine (DA) and/or serotonin (5-HT) terminals. Animals were given a neurotoxic regimen of mAMPH, p-chloroamphetamine (PCA, preferentially damages 5-HT terminals), d-amphetamine (d-AMPH, preferentially damages DA terminals), or saline. After 1 week, animals were trained and tested for OR memory. Rats treated with mAMPH showed no recognition memory during the short-term memory (STM) test, whereas both PCA- and d-AMPH-treated rats showed OR STM scores comparable to controls. After behavioral testing, the specificity of monoaminergic lesions was determined by postmortem [125I]RTI-55 binding to dopamine (DAT) and serotonin (SERT) transporter proteins. Tissue from a separate group of animals killed 3 days after drug treatment was processed for Fluoro-Jade (F-J) fluorescence histochemistry to detect damaged cortical neurons. mAMPH treated rats showed reductions in striatal DAT and hippocampal (HC) and perirhinal (pRh) SERT, as well as degeneration of neurons in primary somatosensory cortex. In PCA-treated rats, HC and pRh SERT were substantially depleted, but striatal DAT and cortical neuron survival were unaffected. By contrast, d-AMPH-treated animals showed marked depletions in striatal DAT and cortical neurodegeneration, but HC and pRh SERT were unaffected. This pattern of results indicates that no single feature of mAMPH-induced neurotoxicity is sufficient to produce the OR impairments seen after mAMPH treatment. PMID- 15900318 TI - N-desmethylclozapine, a major metabolite of clozapine, increases cortical acetylcholine and dopamine release in vivo via stimulation of M1 muscarinic receptors. AB - The active moiety of clozapine, the prototypical antipsychotic drug, consists of clozapine and its major metabolite, N-desmethylclozapine (NDMC). Previous studies have suggested that NDMC may be more important than the patent compound itself for the improvement in cognition in patients with schizophrenia treated with clozapine. While the pharmacology of clozapine and NDMC are similar in most respects, NDMC has been shown to be an M1 muscarinic receptor partial agonist whereas clozapine is an M1 antagonist in vitro and in vivo. We hypothesized that NDMC may improve cognition by increasing dopamine (DA) and acetylcholine (ACh) release in medial prefrontal cortex (mPFC) via direct stimulation of M1 receptors, whereas both NDMC and clozapine itself would do so by other mechanisms as well, and that clozapine would inhibit the M1 agonist effect of NDMC. In the present study, using microdialysis in awake, freely moving rats, we found that NDMC at doses of 10 and 20, but not 5 mg/kg, significantly increased DA and ACh release in the mPFC and HIP, but not in the nucleus accumbens (NAC). The M1 preferring antagonist, telenzepine (3 mg/kg), completely blocked NDMC (10 mg/kg) induced increases in cortical DA and ACh release. Clozapine (1.25 mg/kg), which by itself had no effect on DA or ACh release in the cortex, blocked NDMC (10 mg/kg)-induced ACh, but not DA, release in the mPFC. The 5-HT1A receptor antagonist, WAY100635 (0.2 mg/kg) blocked NDMC (20 mg/kg)-induced cortical DA but not ACh release. These findings suggest that: (1) NDMC is an M1 agonist while clozapine is an M1 antagonist in vivo; (2) M1 agonism of NDMC can contribute to the release of cortical ACh and DA release; (3) NDMC, because of its M1 agonism, may more effectively treat the cognitive impairments observed in schizophrenia than clozapine itself; and (4) M1 receptor agonism may be a valuable target for the development of drugs that can improve cognitive deficit in schizophrenia, and perhaps other neuropsychiatric disorders as well. PMID- 15900319 TI - Inactivation of the cerebral NFkappaB pathway inhibits interleukin-1beta-induced sickness behavior and c-Fos expression in various brain nuclei. AB - The behavioral effects of peripherally administered interleukin-1beta (IL-1beta) are mediated by the production of cytokines and other proinflammatory mediators at the level of the blood-brain interface and by activation of neural pathway. To assess whether this action is mediated by NFkappaB activation, rats were injected into the lateral ventricle of the brain with a specific inhibitor of NFkappaB activation, the NEMO Binding Domain (NBD) peptide that has been shown previously to abolish completely IL-1beta-induced NFkappaB activation and Cox-2 synthesis in the brain microvasculature. NFkappaB pathway inactivation significantly blocked the behavioral effects of intraperitoneally administered IL-1beta in the form of social withdrawal and decreased food intake, and dramatically reduced IL-1beta induced c-Fos expression in various brain regions as paraventricular nucleus, supraoptic nucleus, and lateral part of the central amygdala. These findings strongly support the hypothesis that IL-1beta-induced NFkappaB activation at the blood-brain interface is a crucial step in the transmission of immune signals from the periphery to the brain that underlies further events responsible of sickness behavior. PMID- 15900320 TI - Current outlook of cardiology. Interview. PMID- 15900321 TI - Artificial metallonucleases. AB - The development of synthetic agents able to hydrolytically cleave DNA with high efficiency and selectivity is a fascinating challenge that will show the way to obtaining artificial nucleases able to compete with the natural enzymes. This Feature Article highlights the progress reported toward the realization of synthetic nucleases with particular attention to the strategies that can be pursued to improve efficiency and sequence selectivity. PMID- 15900322 TI - Inhibition of matrix metalloproteinase-9 by "multi-prong" surface binding groups. AB - A novel strategy of blocking the active site accessibility of MMP-9 by "multi prong" surface binding groups is described. PMID- 15900323 TI - Synthesis of constrained helical peptides by thioether ligation: application to analogs of gp41. AB - We present a straightforward and high yielding method to synthesize constrained helical peptides via thioether ligation; this method represents an attractive alternative to the formation of lactam bridge constraints to induce helicity in peptides. PMID- 15900324 TI - Recognition of CG inversions in DNA triple helices by methylated 3H-pyrrolo[2,3 d]pyrimidin-2(7H)-one nucleoside analogues. AB - Substituted 3H-pyrrolo[2,3-d]pyrimidin-2(7H)-one nucleoside analogues have been synthesised from 5-alkynyl-uridine derivatives, incorporated into triplex forming oligonucleotides (TFOs) and found to selectively bind CG inversions with enhanced affinity compared to T. PMID- 15900325 TI - Biosynthesis of conjugatable saccharidic moieties of GM2 and GM3 gangliosides by engineered E. coli. AB - Oligosaccharidic moieties of GM(2) and GM(3) gangliosides bearing an allyl or a propargyl aglycon, are efficiently biosynthesized on the gram scale by growing metabolically engineered Escherichia coli cells in the presence of the corresponding lactoside acceptors and sialic acid. PMID- 15900326 TI - Recognition of homo-polynucleotides containing adenine by a phenanthridinium bis uracil conjugate in aqueous media. AB - Among novel bis-nucleobase-phenanthridinium conjugates bis-uracil analogue stabilized significantly more effective poly-dA-poly-dT and poly-AH(+)-poly-AH(+) than adenine analogue and reference compound . For the alternating poly-dAdT-poly dAdT however, the binding preference is lost, pointing to the importance of specific interactions of uracils of with homopolynucleotides containing consecutive adenines. PMID- 15900327 TI - Enforcing solution phase nanoscopic aggregation in a palindromic tripeptide. AB - C-terminal dimerization of a tripeptide palindrome afforded fibrillation in solution through an assembly probably driven by hydrogen bonding and hydrophobic contributions; such an approach provides an expeditious entry into fabrication of fibrillating peptides from non-fibrillating peptide sequences. PMID- 15900328 TI - Hydrogenation reactions using scCO2 as a solvent in microchannel reactors. AB - We have developed an effective microfluidic system for hydrogenation reactions in scCO(2); the reactions proceeded very rapidly (within 1 second), by making the best use of scCO(2) and utilizing the large specific interfacial area of the microchannel reactor, and high reaction productivity was attained in each channel. PMID- 15900329 TI - Complementarity in bimolecular photochromism. AB - Irradiating 2,3,6,7-tetraphenylanthracene in the presence of 9,10 dimethylanthracene leads to exclusive formation of the cross-dimer. No photochemical reaction is observed when either of these chromophores is irradiated in the absence of the other. PMID- 15900330 TI - Stereo-differentiation in the excited state behaviour of naphthalene-thymine dyads. AB - Using two diastereomeric dyads containing naphthalene and thymine units, significant chiral discrimination has been found in the photophysical processes involving the naphthalene excited states: singlet deactivation by hydrogen bonding molecules, singlet-singlet energy transfer from thymine and triplet decay. PMID- 15900331 TI - Epoxytwinol A, a novel unique angiogenesis inhibitor with C2 symmetry, produced by a fungus. AB - We isolated a novel unique pentaketide dimer designated as epoxytwinol A from the fermentation broth of a fungus. The structure of epoxytwinol A was determined to have a new carbon skeleton with C(2) symmetry by elucidation of spectroscopic evidence. Epoxytwinol A inhibited endothelial cell migration stimulated by vascular endothelial growth factor (ED(100)= 2.6 microM). PMID- 15900332 TI - Kinetic analysis of hydrolytic reaction of homo- and heterochiral adenylyl(3' 5')adenosine isomers: breaking homochirality reduces hydrolytic stability of RNA. AB - The hydrolytic stability of the diastereomeric isomers of ApA was compared and the results show that heterochiral ApAs are more rapidly hydrolyzed than homochiral ApAs at low temperatures, suggesting that hydrolytic selection in cold environments in conjunction with selective polymerization may have been effective in enriching the homochirality of RNA. PMID- 15900333 TI - A novel one-pot three-component synthesis of 3-halofurans and sequential Suzuki coupling. AB - A novel sequence of Sonogashira coupling and electrophilic addition to an ynone, with concomitant deprotection and cyclocondensation, opens a new one-pot synthesis of 3-halofurans; the method can be readily elaborated to a new sequential Sonogashira-addition-cyclocondensation-Suzuki reaction to furnish 2,3,5-trisubstituted furans in a one-pot fashion. PMID- 15900334 TI - Direct electrochemistry and electrocatalysis with hemoglobin in water-soluble quantum dots film on glassy carbon electrode. AB - The direct electrochemistry of hemoglobin can be performed by immobilizing hemoglobin in a water-soluble quantum dots (CdSe-ZnS) film on glassy carbon electrode. PMID- 15900335 TI - An efficient solid phase synthesis of 5'-phosphodiester and phosphoramidate monoester nucleoside analogues. AB - An easy and efficient strategy to obtain libraries of 5'-phosphodiester and 5' phosphoramidate monoester nucleoside analogues in a highly pure form has been developed, starting from a new nucleoside based solid support. The nucleoside scaffold has been anchored through a 5'-phosphodiester linkage to Tentagel HL resin, functionalized with a 3-chloro-4-hydroxyphenylacetic linker. The solid phase synthesis of small libraries of 5'-phosphodiester and 5'-phosphoramidate monoester thymidine analogues is also reported. PMID- 15900336 TI - Aryl-aryl coupling via directed lithiation and oxidation. AB - Aryl lithium reagents formed by directed lithiation reactions undergo transmetallation with copper(I) salts to form organocuprates, which may be efficiently oxidized to yield ortho-substituted biaryls. PMID- 15900337 TI - Carbon-nitrogen bond cleavage in pyridine ring systems mediated by organometallic thorium(IV) complexes. AB - Thorium(IV) alkyl and aryl complexes of the type (C(5)Me(5))(2)ThR(2)(R = CH(2)Ph, Ph) have been found to mediate the facile ring-opening and dearomatization of the pyridine ring of pyridine N-oxide under ambient conditions to afford the first thorium eta(2)-(O,N)-oximate complexes. PMID- 15900338 TI - Elucidating interactions of ionic liquids with polymer films using confocal Raman spectroscopy. AB - We report on the molecular interactions between room-temperature ionic liquids (RTILs) and Nafion and PDMS membranes, proving that in contact with these polymers RTILs behave like electrolytes rather than solvents. PMID- 15900339 TI - Stereoselective hydroxylation of an achiral cyclopentanecarboxylic acid derivative using engineered P450s BM-3. AB - Substrate engineered, achiral carboxylic acid derivative was biohydroxylated with various mutants of cytochrome P450 BM-3 to give two out of the four possible diastereoisomers in high de and ee. The BM-3 mutants exhibit up to 9200 total turnovers for hydroxylation of the engineered substrate, which without the protecting group is not transformed by this enzyme. PMID- 15900340 TI - New hybrid organic-inorganic nanocomposites based on functional [Ti16O16(OEt)24(OEMA)8] nano-fillers. AB - New hybrid nanocomposites based on a methacrylate functionalized titanium-oxo cluster as nano-cross-linker show improved mechanical properties, optical transparency and photochromic activity. PMID- 15900341 TI - Enhancement of oxygen and methane solubility in 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl) imide using carbon dioxide. AB - The presence of CO(2) increases the solubility of O(2) and CH(4) in 1-hexyl-3 methylimidazolium bis(trifluoromethylsulfonyl) imide at 25 degrees C and pressures to 13 bar. PMID- 15900342 TI - Hindered rotation in an "exploded" biphenyl. AB - The first cases of hindered rotation around the triple bond in simple diphenylacetylenes were observed, including that in chiral 2,2' bis(trimethylsilyl)-6,6'-bis(dimethylthexylsilyl)diphenylacetylene. PMID- 15900343 TI - Fabrication of highly porous and micropatterned SnO2 films by oxygen bubbles generated on the anode electrode. AB - The fabrication process of highly porous SnO(2) thick film by reaction between tin ions and oxygen gas generated by an anodic applied potential on substrates in SnCl(2) aqueous solution is reported; moreover, we succeeded in forming porous SnO(2) micropatterns through site-selective deposition on a Pt-patterned F-doped SnO(2)(FTO) coated substrate . PMID- 15900344 TI - meso-Dichloropyrimidinyl substituted expanded porphyrins. AB - Condensation of 4,6-dichloro-2-phenylpyrimidine-5-carbaldehyde with pyrrole enables the synthesis of a number of expanded porphyrins of which the [26]hexaphyrin was easily isolated and successfully converted into the decasubstituted doubly N-fused hexaphyrin variant. PMID- 15900345 TI - Selenoaroyl complexes of molybdenum. AB - The reaction of mesityl isoselenocyanate with molybdenum alkylidynes provides the first structurally characterised examples of mononuclear selenoaroyl complexes, which may also be obtained directly from elemental selenium in the presence of a catalytic amount of mesityl isocyanide. PMID- 15900346 TI - Synthesis of ordered macroporous SiO2 in supercritical CO2 using 3D-latex array templates. AB - Ordered macroporous SiO(2) membranes were produced by the decomposition of silicon alkoxides in supercritical carbon dioxide (scCO(2)) using 3D-latex arrays as templates. PMID- 15900347 TI - TiCl4 catalyzed tandem construction of C-C and C-O bonds: a simple and one-pot atom-economical stereoselective synthesis of spiro-oxindoles. AB - An atom-economical stereoselective synthesis of [{1-acetyl-5-methyl-6,8 dioxabicyclo(3.2.1)octane}-7-spiro-3'-(indolin-2'-one)] derivatives, containing both the oxindole and 6,8-dioxabicyclo(3.2.1)octane moieties via TiCl(4) catalyzed coupling of 2-acetyl-6-methyl-2,3-dihydro-4H-pyran with isatin derivatives is described. PMID- 15900348 TI - The rhodium catalyzed three-component reaction of diazoacetates, titanium(IV) alkoxides and aldehydes. AB - The rhodium(II)-catalyzed three-component reaction of diazoacetates, titanium alkoxides and aldehydes is shown to give alpha-alkoxyl-beta-hydroxyl acid derivatives; the novel C-C bond formation reaction is proposed to occur through oxonium ylides derived from diazo compounds and titanium alkoxides, and followed by intermolecular trapping by aldehydes. PMID- 15900349 TI - Sustained production of H2O2 on irradiated TiO2- fluoride systems. AB - UV irradiation of fluorinated TiO(2) suspensions in water, in the presence of oxygen and a hole scavenger, leads to the production of H(2)O(2) with steady state concentration levels up to 1.3 millimolar; the H(2)O(2) formation rate follows the TiO(2) surface speciation, being maximum when the surface is completely covered by [triple bond]Ti-F groups; these results outline the importance of surface speciation on the photocatalytic process. PMID- 15900350 TI - A zinc(II)-based receptor for ATP binding and hydrolysis. AB - A protonated Zn(II) complex with a terpyridine-containing pentaamine macrocycle catalyses ATP hydrolysis in the presence of a second metal ion, which acts as cofactor assisting the phosphoryl transfer from ATP to an amine group of the receptor. PMID- 15900351 TI - One pot synthesis of 3,5-alkylated acetophenone and methyl benzoate derivatives via an anionic domino process. AB - The reaction of primary 1,3-dinitroalkanes with 2-ene-1,4-dione or 2-ene-4-oxo ester derivatives in acetonitrile with DBU as base, allow the one pot synthesis of 3,5-alkylated acetophenones and methyl benzoate derivatives respectively via an anionic domino process. PMID- 15900353 TI - Helicobacter pylori. Basics and clinical overview. AB - The discovery of Helicobacter pylori (H. pylori) has greatly changed the approach to the management of peptic ulcer disease and gastric cancer. A sound knowledge of the basics of H. pylori is an important aid in the diagnosis and treatment of clinical conditions associated with this infection. Gastric carcinoma is estimated to be the world's second most common cancer as a cause of death. It is hoped that gastric cancer can be prevented by H. pylori eradication; however, this issue is still under investigation. Active research is ongoing to highlight the mechanisms by which H. pylori leads to severe gastric diseases as well as finding associations with extra-gastric diseases. PMID- 15900354 TI - Correlation between developmental stages of the human heart and gestational ages. AB - OBJECTIVES: In this study we aimed to investigate the normal developmental pattern of the human fetal heart in second and third trimester, and to evaluate the correlation of the values with the gestational age (GA). METHODS: Seventy spontaneously aborted human fetuses with no external malformations were investigated and 11 parameters from each were assessed. The relationship of these parameters with the GA was evaluated by linear regression and correlation analyses using statistical methods. In addition, the values were compared to the gender and sides of the heart using Student t-test. This study was performed at the Department of Anatomy, Meram Medical Faculty, Selcuk University, Konya, Turkey, during the period from 2000 to 2004. RESULTS: There was a statistically significant linear relationship between the heart wall thicknesses, the development of the valves of the heart and GA. The mean values of the right side of the heart were found to be higher than those of the left side, and differences were statistically significant (p<0.05). Although, most of the parameters were found to be higher in females, only 2 of them were statistically significant. All parameters of the cardiac valves measured were significantly higher in the second trimester. There was no difference regarding the correlation ratio of the trimesters between the pulmonary valve diameter and GA. The measurement of the cardiac valves did not show any differences regarding the gender. CONCLUSION: This study presents the normal values of the development of the fetal heart in second and third trimester that may provide useful background information for clinical approaches. PMID- 15900355 TI - Khat chewing and arterial blood pressure. A randomized controlled clinical trial of alpha-1 and selective beta-1 adrenoceptor blockade. AB - OBJECTIVE: The aim of this work was to study prospectively the effect of alpha-1 and selective beta-1 adrenoceptor blockade on cardiovascular effects of khat chewing in healthy volunteers. METHODS: Sixty-three male volunteers chewed khat for 3 hours on 3 separate occasions, 1 or 2 weeks apart. Fifty age and weight matched male controls did not chew. The khat chewers received in a double blind 3 arm crossover design either indoramin 25 mg, atenolol 50 mg or placebo one hour before starting to chew. The non-chewing controls received atenolol 50 mg on a separate occasion. Pulse and blood pressure were measured at regular intervals before, during and after the chewing period. The study was carried out in Sana'a, Yemen between December 2001 and November 2003. RESULTS: Khat chewers in the atenolol treated group had significantly lower readings for systolic blood pressure (SBP) and pulse rate one, 2 and 3 hours after starting to chew than khat chewers pre-treated with placebo or indoramin and comparable to non-khat chewers. Three hour SPB readings in khat chewers with placebo, with atenolol and with indoramin (mean values [95% confidence interval]) were 123 (120.2-125.7), 115.7 (113.0-118.4), and 119.8 (116.9-122.8) (p<0.0001). Blood pressure and pulse rate were not altered in non-khat chewers taking 50 mg atenolol. Diastolic blood pressure during khat chewing rose in the atenolol, indoramin and placebo groups. CONCLUSION: The effect of khat chewing on systolic blood pressure and pulse rate is blocked by atenolol but not by indoramin. Beta-1 adrenoreceptors are probably important in mediating the cardiovascular effects of khat in man. PMID- 15900356 TI - Factors determining self-efficacy in chronic obstructive pulmonary disease. AB - OBJECTIVE: We investigated the factors determining self-efficacy in patients with chronic obstructive pulmonary disease (COPD). METHODS: Thirty male patients with COPD and 30 healthy controls participated in the study. Pulmonary function tests, 6-minute walk test (6MWT), modified Borg scale, modified Medical Research Council (MRC) dyspnea scale, Hospital Anxiety and Depression Scale (HADS), Activities of Daily Living Questionnaire (ADL-Q), and COPD Self-Efficacy Scale (CSES) were used for the evaluation. The study was performed between September 2003 and May 2004 at the Department of Chest Medicine, School of Physical Therapy and Rehabilitation, Hacettepe University, Ankara, Turkey. RESULTS: The COPD patients expressed higher depression and dyspnea level and impaired activities of daily living than healthy controls (p<0.05). Duration of disease and the ADL-Q score was significantly correlated with all subscales of CSES (p<0.05). Borg score and anxiety score were significantly related with negative affect, weather/environment, and behavioral risk factors subscales of CSES (p<0.05). Lung function was significantly related with physical exertion and behavioral risk factors, and 6MWT distance was significantly correlated with weather/environment and behavioral risk factors subscales (p<0.05). The MRC score was correlated with weather/environment score (p<0.05). The ADL-Q score and disease duration accounted for 76% of the variance in self-efficacy (p<0.05) CONCLUSION: Level of impaired activities of daily living due to respiratory limitation and the number of years since diagnosis are independent variables that predict self-efficacy, or level of confidence in engaging specific behaviors that lead to specific desired outcomes in COPD. PMID- 15900357 TI - Extent of potential drug interactions among patients receiving anti-hypertensive medications. AB - OBJECTIVE: To investigate the frequency of potential antihypertensive drug interactions among patients with cardiovascular diseases receiving antihypertensive medications. METHODS: The study took place in Nablus, Palestine starting April through October 2003. Patients with cardiovascular diseases (n=876) or who were receiving one or more antihypertensive medications were evaluated. All drugs prescribed for the patients were obtained from their medical files. A drug interaction database was developed based on updated Drug Interaction Facts to examine potential and level of drug interactions in each patient's regimen. Data were entered and analyzed using SPSS software. RESULTS: The number of "unique" pairs of potential drug interactions among the antihypertensive agents present in the data was 433. These included 16 cases (3.7%) level one; 34 cases (7.8%) level 2; 116 cases (26.8%) level 3; 136 cases (31.4%) level 4, and 131 (30.3%) level 5 interactions. Both increasing age and number of drugs were significantly associated with the potential for significant interactions at all levels with a p value less than 0.025. CONCLUSIONS: This study found a high frequency of potential drug interactions with agents typically used for hypertension. Similar investigations need to be carried out among patients with other types of chronic diseases. Drug interaction software might be necessary in governmental pharmacy departments. PMID- 15900358 TI - Comparison between drainage and curettage in the treatment of acute pilonidal abscess. AB - OBJECTIVE: Simple incision and drainage of a sacrococcygeal pilonidal abscess is associated with more than 40% recurrence. Definitive treatment of the chronic pilonidal sinus is recommended 4-8 weeks after incision and drainage. The present prospective clinical trial study was designed to compare the curettage and drainage of the acute abscess of the pilonidal sinus. Hospital stay, wound healing, and recurrence were noted. METHODS: This study was performed in Kerman University of Medical Sciences and Health Services, Iran from March 1999 to May 2004. One hundred and fifty patients who had an acute sacrococcygeal pilonidal abscess were randomly assigned to receive one of 2 drainage or curettage surgical procedures. All the surgical wounds were laid open, and daily sitting in a warm tub together with douche was recommended postoperatively. The 2 treatment modalities were compared in terms of the wound healing period, hospital stay, and postoperative complications using Chi-square and Student-t test. Patients with completed wound repair, consisting of 72 subjects in the curettage and 59 subjects in the drainage groups, were followed up to 65 months for the detection of recurrence by means of life table and Gehan s generalized Wilcoxon test. RESULTS: Most patients spent only one day in the hospital, with initial healing occurring in <2 months. Healing process in the curettage cases occurred more quickly than the drainage patients (96% versus 78.7%, p<0.001). In all cases, the treatment relieved symptoms, and all the patients returned to work 7-10 days after the treatment. Three patients in the curettage group (4%) and 16 patients in the drainage group (21.3%) did not complete wound healing within 10 weeks (p<0.001). Recurrence of disease was encountered significantly less in the curettage subjects than the drainage ones (11% versus 42%) (p<0.001). CONCLUSION: Unroofing and curettage, which is associated with higher rates of healing and lower rates of recurrence, may be the treatment of choice in the acute pilonidal abscess. PMID- 15900359 TI - Laparoscopic appendectomy in children. AB - OBJECTIVES: Laparoscopic appendectomy (LA) is rapidly becoming an alternative to open appendectomy for the treatment of appendicitis in children. We examined the efficacy, safety and complications of performing LA in children. METHODS: This is a critical review of the children who underwent LA between January 1999 and May 2003 at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. Demographic data, operative procedures, severity of appendicitis, histopathology, operative time, hospital stay, analgesia and complications were carefully analyzed. RESULTS: Laparoscopic appendectomy was attempted in 113 children. It was successfully completed in only 100 patients, 85 had uncomplicated appendicitis and 15 had complicated appendicitis. In 13 (11.5%) conversion to open appendectomy had to be carried out for complicated appendicitis. Their ages ranged between 2-12 years (average 8 years); 52 were males and 48 were females. Symptom duration average was 30 hours in uncomplicated appendicitis and 56 hours in complicated appendicitis. Operative time ranged between 35-80 minutes (average 53 minutes). Four patients with complicated appendicitis developed postoperative complications, 2 had wound infections and the other 2 had intra-abdominal collections. No intra-operative complications were encountered. The average hospital stay was 2 days for uncomplicated appendicitis and 6 days for complicated ones. Histopathological examination revealed 75 acutely inflamed appendices, 15 were perforated appendicitis with or without mass formation and 10 appendices reported to be normal. All patients were followed up for 6-12 months. CONCLUSION: Laparoscopic appendectomy is both a safe and effective method in managing children with acute uncomplicated appendicitis. In children with complicated appendicitis, LA should be carried out with caution and only by an experienced surgeon, as it is associated with a higher rate of conversion to open technique and more postoperative infectious complications. PMID- 15900360 TI - Birth anthropometric parameters in high and low altitude areas of Southwest Saudi Arabia. AB - OBJECTIVES: To study the anthropometric parameters of Southwestern Saudi newborns in Abha (a high altitude area) and Baish (a sea level area of Southwestern Saudi Arabia); to compare these parameters with those of the United States Center for Disease Control 2000 growth charts and to estimate the incidence of low birth weight (LBW) among the study population. METHODS: The study sample included all births from Abha Central Hospital (N=4300), and all births from Baish General Hospital (N=1200), Kingdom of Saudi Arabia during the period from January 2001 to January 2003. Only term births (>37-42 weeks gestation) were included. The anthropometric measurements included birthweight, crown-heel length and head circumference. The head circumference (occipitofrontal) was measured to the nearest 5 mm with an inelastic tape. Babies of LBW were identified. Low birth weight was defined as <2,500 gms at birth. Ponderal index (PI) was calculated using the formula of birthweight (in grams) x 100/length3. RESULTS: Saudi newborns are lighter and shorter than those of National Center for Health Statistics newborns. The same finding was evident for head circumference. Newborns from Abha are lighter (p<0.001) and shorter (p<0.001) than from Baish. However, this difference was not significant for head circumference (p=0.53). Abha newborns showed significantly lower mean weight (2845.4 gms versus 2951.8 gms, t=58, p<0.001), and lower mean length (48.1 cm versus 48.5 cm, t=4.65, p<0.001). Low birth weight was prevalent among 24.6% of all newborns. This figure was significantly higher among newborns from Abha (25.7%) than its counterpart in Baish (20.7%), p<0.001. A strong positive correlation between the PI and birthweight (p<0.0001, R2=0.52), after adjusting for the place of birth was evident. CONCLUSION: Neonates of Southwestern Saudi Arabia are significantly lighter and shorter than those of the reference population, as well as neonates in other areas of the Kingdom, with prevalent LBW especially in high altitude area, possibly due to intra-uterine growth retardation of secondary type. Ponderal index is significantly associated with birth weight even after adjusting for gender and place of birth. This might encourage the use of PI as an alternative to birth weight especially when gestational age is not available. PMID- 15900361 TI - Pattern of headache in school children in the State of Qatar. AB - OBJECTIVES: The aim of this study is to determine the prevalence rate and impact of headache in school children on school attendance with particular attention to migraine. METHODS: A cross sectional survey was conducted among the school children ranging from 6-17 years old over a period from March 2001 to April 2003. The study was carried out in 10 primary, preparatory and secondary schools. Subjects were selected by multistage stratified sampling procedure. This involved 851 children studying in the first to tenth year of school in the State of Qatar. RESULTS: The present study showed that the prevalence rate of recurrent headache was 85% and migraine 11.9%. Comparing gender frequency of headache, it was noted that it was higher in female students (86.5%) than males (81%). In respect of age, it was observed that the oldest children had more frequent episodes of headache, the highest rate was in the age group of 11-15 years old (49%). The most common triggers were fatigue (35.8%) and lack of sleep (17.6%). The most common symptoms that occurred before headache were change in mood for female students (39.1%) and blurred vision for males (34.6%). The impact of headache on children was frequent absence from school (80%), which affected their school performance. CONCLUSION: The current study indicated the high prevalence of headache among school children in Qatar, and its effect on school attendance and performance. PMID- 15900362 TI - Hodgkin's disease and association with Epstein-Barr virus in children in Southeast Turkey. AB - OBJECTIVES: There are relatively few reports on histologic and immunophenotypic features of Hodgkin's disease (HD) in children in Turkey. The aim of the present study is to characterize the clinicopathologic, immunophenotypic features and Ebstein-Barr virus (EBV) status of HD in children in our region. METHODS: Fifty two cases coded as HD in the Department of Pathology, Medical Faculty, Dicle University, Turkey, from 1990 to 2002 were retrieved. We analyzed clinicopathologic data, immunophenotype, and EBV status of all patients. RESULTS: In most cases, the cervical lymph nodes were involved. The 52 cases (35 boys, 17 girls; male to female ratio was 2.1) were categorized as mixed cellularity (MC) in 23 (44.2%), lymphocyte predominance (LP) in 16 (30.8%), nodular sclerosis (NS) in 10 (19.2%), and lymphocyte depletion (LD) in 3 (5.8%). Of 33 cases, including MC and NS, Hodgkin's Reed-Sternberg cells and variants were positive for CD15 in 27 cases (81.8%) and positive for CD30 in 30 cases (90.9%). All the cases of LP demonstrated the characteristic phenotype of this variant (CD45RB+, CD20+, CD15-, CD30-). Epithelial membrane antigen stained the lymphocytic/histiocytic cells in 3 cases (18.7%) of 16. Epstein-Barr virus-LMP1 were positive in 31 cases (61.5%) of a total of 52 cases and the most were MC (91.3%). CONCLUSION: In conclusion, mixed cellularity HD is the most common histologic subtype. This result differed from that reported in developed countries. We found a striking association with expression of EBV-latent membrane protein 1 in malignant cells in childhood HD. PMID- 15900363 TI - Gastritis in Saudi Arab children. AB - OBJECTIVE: Information on childhood gastritis in developing countries is rare; hence, the objective of this study is to report the pattern of this condition in Saudi Arab children. METHODS: Data analysis were carried out in all children <18 years of age who were referred for endoscopy at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia from 1993 to 2002. Only children with histology proven gastritis were included. RESULTS: One hundred and seventy-five patients out of 851, referred for the investigation of gastrointestinal symptoms, had histology-proven gastritis; whereas only 110/851 (13%) had endoscopic features of gastritis. Most of the children (96%) were Saudi nationals, the age ranged between 4 days to 18 years, and the male to female ratio was 0.4:1. The most common presentation of gastritis was abdominal pain in 42%, followed by combination of abdominal pain and vomiting in 20% and vomiting alone in 18% of the children. Rare but important presentations included refractory anemia, melena and halitosis. The prevalence of gastritis increased with age from 9% in children <5 years of age to 42% in adolescents 16-18 years of age. Likewise, the prevalence of Helicobacter pylori (H. pylori) gastritis increased from 47% in children <5, to 62% in the 6-15 year age group, and 69% in the 16-18 years old adolescents. Endoscopy was normal in 74/175 (42%) of histologic gastritis and the prevalence of H. pylori gastritis was the highest in cases of endoscopic nodularity (92%) CONCLUSION: Compared to findings from various parts of the world, this report document a similar clinical pattern, but high prevalence of H. pylori gastritis in Saudi Arab children. PMID- 15900364 TI - Prevalence of factor V Leiden and prothrombin G20210A gene mutation. AB - OBJECTIVES: To determine the prevalence of factor V Leiden (FVL) and prothrombin gene (PG) 20210A mutations in patients who attended the outpatient clinic and do not have a family history of thrombosis. METHODS: We researched FVL and PG20210A mutations in 151 outpatients (92 males and 59 females) who attended the Pediatrics Polyclinic, Medical Faculty, Dicle University, Turkey between May 2002 and July 2002. Peripheral venous bloods (2 cc) with ethylenediaminetetraacetic acid were used to isolate DNA by high pure polymerase chain reaction (PCR). Later, by using light-cycler FVL (Roche) and prothrombin mutation detection kit (Roche) and light-cycler equipment, FVL and PG20210A gene mutations were determined from the samples in the glass capillary tubes by PCR specific adaptation. For active protein C (APC) resistance, STA-STACLOT APC-R Detection Kit was used. Expected values were evaluated with STA equipment and using STA STACLOT APC-R procedure. RESULTS: The prevalence of heterozygote mutations of FVL was 4.6% and PG20210A was 0.7%. The FVL mutation frequency obtained in our study is lower than the other studies in Turkey, but in correlation with the results of the other Caucasian populations throughout the world. Active protein C resistance in patients carrying heterozygote mutation of FVL has been found in low rates. Factor V Leiden and PG20210A were confronted in high prevalences in patients who suffer venous thrombosis (VT) CONCLUSION: Scanning of FVL and PG20210A gene mutations may be recommended in high risk groups such as relatives of FVL and PG20210A carriers and relatives of patients with VT, and in during pregnancy, the use of oral contraceptives and before surgery. Routine scanning of FVL and PG20210A gene mutations is not recommended in people who do not have risk factors for VT. PMID- 15900365 TI - Accuracy of current oxacillin sensitivity tests routinely used in hospitals in Western Saudi Arabia. AB - OBJECTIVE: To determine the accuracy of the current oxacillin resistant Staphylococcus aureus (S. aureus) detection test used in Makkah hospitals in comparison with the National Committee for Clinical Laboratory Standards (NCCLS) method. METHODS: A total of 500 S. aureus clinical isolates and it's oxacillin sensitivity patterns were collected from the 4 main hospitals in Makkah, Kingdom of Saudi Arabia between April 2003 and January 2004. The oxacillin sensitivity of these clinical isolates were re-examined using the NCCLS standard method and confirmed using polymerase chain reaction (PCR) technique. RESULTS: Of 500 clinical isolates, 103 (20.6%) were resistant to oxacillin using NCCLS standard method but they were sensitive according to the current hospital routine sensitivity test method. The PCR technique confirmed the presence of mecA gene in 88/103 isolates appeared to be methicillin-resistant S. aureus (MRSA) using NCCLS standard technique. CONCLUSION: A significant percentage of MRSA are currently misdiagnosed in accordance with the current routine sensitivity method. In addition, some mecA negative and oxacillin resistant strains (according to the NCCLS standard method) can be misdiagnosed by using PCR technique. These findings emphasis the urgent need to comply with the recommended NCCLS guidelines for detection of oxacillin resistance. Moreover, the PCR technique can not be used as a single diagnostic tool for detection of MRSA. PMID- 15900366 TI - Cytopathological pattern of cervical Pap smear according to the Bethesda system in Southwestern Saudi Arabia. AB - OBJECTIVE: Cervical carcinoma is an important women's health problem worldwide. The aim of this study is to evaluate the abnormal cytological entities detected by cervical Pap smear in Al-Baha region of KSA, to reclassify the cytological findings according to the recently recommended Bethesda system and to discuss the significance of Pap smear's screening programs. METHODS: A retrospective study was designed to review the Pap smears from the archives of the Department of Pathology, King Fahad Hospital, Al-Baha over the last 10 years starting from 1994 to 2003 with correlation to the Bethesda system. Cytopathological aspects of Pap smears were reviewed with age distribution. RESULTS: Of the 2100 cases reviewed there were 166 cases (7.9%) with epithelial abnormalities that included: atypical squamous cells of undeterminate significance (ASCUS) were seen in 58 cases (2.76%) with age peak incidence (API) between 30-39 years; atypical squamous epithelium, cannot exclude high squamous intraepithelial lesion (HSIL) was seen in 4 cases (0.19%); low grade squamous intraepithelial lesion (LSIL) including human papillomavirus was seen in 27 cases (1.3%) with API between 50-59 years and HSIL was seen in 14 cases (0.66%) with API between 40-49 years. Squamous cell carcinoma was seen in 7 cases (0.33%) with API between 50-59 years. There were 54 cases (2.57%) of atypical endocervical cells and 2 cases (0.09%) of atypical endometrial cells. CONCLUSION: Atypical squamous cells of undeterminate significance, LSIL, HSIL, invasive cervical carcinoma and sexually related infectious diseases are less frequently encountered in Saudi women and occur at older age compared to the Western countries. Fewer women are screened for this disease in this region and health education is very important to encourage more Saudi female to have this important screening test particularly in this region of KSA. PMID- 15900367 TI - Evaluation of ovulation induction protocols for poor responders undergoing assisted reproduction techniques. AB - OBJECTIVE: To compare 3 stimulation protocols in poor ovulation responders undergoing in-vitro fertilization (IVF). METHODS: The study was a randomized, prospective clinical trial from June 2003 to July 2004, in Royan Institute, Tehran, Iran. One hundred and fifty-four patients, who had poor responses to ovulation induction in at least one previous IVF attempt, were randomly divided into 3 groups. In the first group, human menopausal gonadotropin (HMG) was administered from day 3 of the cycle at a dose rate of 150 IU/day. In the second group, gonadotropin-releasing hormone (GnRH) agonist was started at a dose rate of 800 microg/day by nasal spray or 500 microg/day subcutaneously in the mid luteal phase, followed by a standard HMG dose after pituitary down regulation was confirmed. In the third group, clomiphene at a dose rate of 100 mg/day was given from day 3 and HMG from day 6. Our main outcomes were number of mature oocytes, cancellation rate, number of HMG ampoules used and incidence premature luteinizing hormone (LH) surge. RESULTS: There was a high incidence of premature LH surge in all groups except in the GnRH group (p=0.0001) and there were significant differences between groups in HMG requirements (p=0.004). There were no significant differences between groups in number of mature oocytes recovered and cancellation rate. CONCLUSION: Results showed no advantage in the use of GnRH agonist compared to the older regimens of clomiphene plus HMG and HMG alone. The cancellation rate was similar for 3 protocols and HMG requirement was higher with the use of GnRH agonist. The treatment of poor responders in assisted reproductive technologies remains a challenge. PMID- 15900368 TI - An investigation of Helicobacter pylori using culture, histopathological and serological examination methods and its antimicrobial sensitivities. AB - OBJECTIVES: In this study, the determination of Helicobacter pylori (H. pylori) by culture, histopathological and serological methods in cases of endoscopically diagnosed as duodenitis and duodenal ulcer (DU), a comparison of their relative advantages, and its antibiotic sensitivities were investigated. METHODS: Helicobacter pylori was investigated using 3 methods (culture, histopathological and serological examination) in 50 patients (25 diagnosed with duodenitis and 25 with DU) at the Department of Gastroenterohepatology, Istanbul Haydarpasa Numune Hospital, Turkey between December 2000 and February 2001. An investigation into its antibiotic sensitivities to amoxicillin, clarithromycin, metronidazole and azithromycin by disc diffusion methods and to amoxicillin and clarithromycin by E test were investigated. RESULTS: Helicobacter pylori bacteria were observed in Gram stained preparates prepared from biopsy material in 34 out of 50 patients (68%), and were able to be produced in active culture in all these cases. Histopathological examination revealed the presence of H. pylori in 80% cases of DU and 60% cases of duodenitis; anti-CagA(IgG) was positively determined in 88% DU cases and in 60% duodenitis cases. There was a significant difference between the 2 groups in terms of diagnosis by histopathological and serological methods. The difference between the 2 groups produced in active culture in 84% cases of DU cases and 52% of duodenitis was statistically significant (p=0.0322). Using the E test and disc diffusion methods, 8.8% of the strains that reproduced in culture were resistant to and 91.2% were sensitive to clarithromycin. All strains were determined to be sensitive to amoxicillin: 17.6% of the strains were determined to be resistant to metronidazole, 11.7% to azithromycin. CONCLUSION: It was observed that Gram staining is a rapid and reliable method of pre-diagnosis for H. pylori; that histopathological examination methods are of considerable importance in diagnosis; and that the investigation of the positivity of anti CagA(IgG) will be a guide in the identification of virulent strains in particular. In addition, it was also concluded that since serological examination does not require invasive measures, this will pose an advantage. The culture method can be applied with the aim of diagnosis in cases identified as DU using endoscopy, and that in cases resistant to treatment it can be applied for the purpose of determining antimicrobial sensitivity. E-test and disc diffusion methods exhibited a rather good correlation, for which reason the disc diffusion method can be used in the determination of antimicrobial sensitivity in H. pylori strains. PMID- 15900369 TI - Occult hepatitis B among chronic liver disease patients. AB - OBJECTIVE: Occult hepatitis B virus (HBV) infection is characterized by presence of HBV infection with undetectable hepatitis B surface antigen (HBsAg). Diagnosis of occult HBV infection requires sensitive HBV-DNA polymerase chain reaction (PCR) assay. Occult hepatitis B is a new entity and the prevalence of it and its clinical importance has not been investigated yet in Iran. The aim of this study is to investigate the prevalence and clinical importance of occult hepatitis B among chronic liver disease patients in Iran. METHODS: We studied 35 consecutive paraffin-embedded liver tissues cases referred to Research Center for Gastroenterology and Liver Diseases, Tehran, Iran during the year 2001 to 2002 for liver biopsy due to its elevation of alanine aminotransferase (ALT) levels for more than 6 months. Liver biopsies were reviewed and HBV-DNA and HBsAg and Hepatitis B core antigen were assayed in liver tissue by PCR and immunohistochemistry (IHC). RESULTS: Our patients with chronic liver disease (CLD) included hepatitis C (77.1%), cryptogenic liver disease (20%), and autoimmune hepatitis (2.9%). Histologically, chronic hepatitis, cirrhosis and non specific changes were reported. Hepatitis B virus-DNA was detectable in 8 (22%) patients; however, IHC was negative in all. CONCLUSION: Occult hepatitis B is relatively frequent among patients with CLD in Iran. It maybe associated with more advanced liver pathology (cirrhosis) and more aggressive clinical course (decompensated cirrhosis). Occult HBV infection causes strong suppression of viral gene expression. PMID- 15900370 TI - Prevalence of obesity and overweight among Saudi adolescents in Eastern Saudi Arabia. AB - OBJECTIVE: To determine the prevalence of obesity and overweight among Saudi adolescents, using the 2 most widely used international references. METHODS: A cross-sectional study conducted towards the end of 2001 on a random sample of third grade intermediate and all 3 grades of secondary school Saudi students of both genders in Al-Khobar area, Eastern Saudi Arabia. The body mass index (BMI) was calculated. The American National Health and Nutrition Examination Survey (NHANES) growth charts, which have been adopted by the World Health Organization (WHO), were used. Student with a BMI of <85th and >95th percentile for age and genders, were defined as overweight and <95th percentile defined as obese. The International Obesity Taskforce (IOTF) age-sex-specific BMI cut-offs reference for defining overweight and obesity was used for comparison. RESULTS: The sample was 1766 students, comprising 675 males and 1091 females. The mean age was 16.4 +/- 1.7 years. The prevalence of obesity was higher in male than female students (19.3% versus 11.8%) while a higher proportion of female students than males were overweight (17.2% versus 10.2%). No significant difference was found between the 2 references used to determine the prevalence of obesity and overweight. CONCLUSION: The high prevalence of overweight and obesity recorded in this study call for prevention programs based on dietary and physical education in schools. PMID- 15900371 TI - Bilateral breast cancer. Incidence, diagnosis and histological patterns. AB - OBJECTIVE: Bilateral breast cancer is uncommon (1-2.6% of all patients with breast carcinoma). There are conflicting reports and inadequate data regarding the incidence and survival of such patients. This study examined the frequency, mode of detection, therapeutic modalities and histological features among the 2 breasts. METHODS: Medical records of patients who were treated for breast cancer were reviewed at Khartoum Teaching Hospital, Sudan during a 5-year period of 1994 to 1999. Data of patients found to have bilateral breast cancer was analyzed focusing on the demographic information, family history, menstrual status, surgical therapy, chemoradiation, staging and histopathological characteristics. RESULTS: Of 521 patients treated for breast cancer, 90 (17.2%) were reported to have advanced breast carcinoma (stage III, IV) and 7 (1.3%) revealed bilateral cancer: 5 (0.9%) synchronous and 2 (0.4%) metachronous breast cancer. The median age was 47.3 years (range, 24-81 years). Four (57.1%) patients had positive family history for breast cancer. First breast cancer was detected by mammogram in 2 cases while second cancer was diagnosed by the same study in 5 patients. Five (71.5%) underwent mastectomy and 2(28.5%) were treated by lumpectomy and axillary dissection. Six patients presented with stage III and one with stage IV breast cancer. Invasive ductal carcinoma was reported to be the most common type found in 10 (71.5%) breasts. Grade III breast carcinoma was revealed in 9 and Grade I cancer in 3 specimens. CONCLUSION: Bilateral breast cancer is invariably advanced when diagnosed. Mammogram is a valuable tool in early detection. Whether synchronous or metachronous, both breasts often share the same histological type. PMID- 15900372 TI - Assessment of sleep patterns, daytime sleepiness, and chronotype during Ramadan in fasting and nonfasting individuals. AB - OBJECTIVE: Sleep changes during Ramadan in nonfasting individuals have not been assessed before. We aimed to assess the effect of the attending lifestyle changes during Ramadan on sleep behavior and chronotype in fasting and nonfasting individuals. METHODS: Participants completed predesigned questionnaires to assess sleep patterns, daytime sleepiness, meal habits, and chronotype one week before Ramadan (BL) and during the first week (R1) and third week (R3) of Ramadan 1424 (Hijra) (between October 26 and November 15, 2003). Participants were divided into to 3 groups: fasting Saudis (SF), fasting non-Saudis (NSF), and non-fasting non-Saudis (NF). The study was conducted in Riyadh, Kingdom of Saudi Arabia. RESULTS: Complete data were available on 41 subjects in the SF group, 30 in the NSF, and 30 in the NF group. There was no changes in total sleep time or daytime sleepiness in all 3 groups. In the fasting groups (SF and NSF), bedtime and wake up time were delayed significantly at R1 and R3 compared to BL, the frequency of meals decreased significantly during Ramadan, and morningness/eveningness test showed an increase in the evening types during Ramadan. In the NF group, bedtime was delayed significantly during Ramadan, and morningness/eveningness test showed an increase in neither types and reduction in the morning types during Ramadan. CONCLUSIONS: Sleep behavior changes during Ramadan in nonfasting participants indicate that the attendant lifestyle changes during Ramadan have a significant effect on sleep behavior. Factors other than fasting may play an important role in modifying an individual's behavior during Ramadan. PMID- 15900373 TI - Health habits and risk factors among Omanis with hypertension. AB - OBJECTIVE: To determine the health habits and risk factors among Omanis with hypertension METHODS: The study was conducted at the Medical Unit of the Royal Hospital, Muscat, Sultanate of Oman during the period from January 2003 to September 2003. A cohort of Omani hypertensive patients (43 male and 57 female) with a mean aged of 51.47 years was randomly selected. All participants were interviewed with a questionnaire for 15-20 minutes. Medical records of the participants were reviewed to determine the presence of associated complications as well as other unrelated chronic health problems based on the diagnosis made by the treating physicians. This was in addition to the measurement and recording of blood pressure, height and weight. RESULTS: The study revealed that the prevalence of uncontrolled hypertension was 73%. The main cause of uncontrolled hypertension was due to unhealthy lifestyle such as more consumption of fatty food and salty food, little physical activity, obesity and age. The study also revealed that there was a significant difference between hypertension and family history, stress, drinking more coffee, physical activity and presence of associate clinical conditions including cardiac and renal. CONCLUSION: Our study shows that lifestyle should be encouraged along with hypertensive therapy as blood pressure control cannot be achieved without reducing salt and fat, maintaining an ideal weight, and maintaining regular exercise. Therefore, it is recommended that the need for effective nursing intervention aimed at increasing levels of awareness of complying with lifestyle modifications, along with hypertensive therapy and family enforcement should be encouraged. PMID- 15900374 TI - Subacute thyroiditis in Western Saudi Arabia. AB - OBJECTIVES: The aim of this study is to assess the clinical presentation of 23 patients with subacute thyroiditis (SAT) and the diagnostic value of radionuclear scan. METHODS: This is a cohort study, which consists of 23 patients with a suspected diagnosis of subacute thyroiditis. The study was carried out in the Endocrinology Clinic, King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia between July 2002 and July 2004. Medical charts including age, gender, clinical presentation, systemic symptoms and clinical examination of the thyroid gland were reviewed. Laboratory data included white blood count and its differential count, erythrocyte sedimentation rate (ESR), thyroid function test and thyroid antibodies. The radionuclear scan results were also noted. The mode of therapy provided to patients and the outcome of the treatment during a follow up period of 2 years was reported. RESULTS: Twenty-three adult patients with subacute thyroiditis (15 females and 8 males with a female to male ratio of 1.9:1) were reviewed over a 2-year period. The mean age was 35.8+/-9.2 years. Eighteen patients (78%) had an upper respiratory tract infection at the initial clinical presentation. Twenty patients (87%) visited an Ear, Nose and Throat specialist for sore throat and abnormal sensation in the throat at least 2 weeks before presentation to the endocrinologist. Two patients were admitted to a medical unit with a diagnosis of fever of unknown origin for 4 weeks. All patients had an elevated free thyroxine (35.7+/-19.8 pmol/L) and suppressed thyroid stimulating hormone (TSH) (0.043+/-0.065 IU). The radionuclear san showed either no uptake at all in 12 patients or minimal uptake in 11 patients (0.32+/ 0.55%). Eight patients (35%) received prednisolone therapy alone with an average dose of 30-40 mg daily for 7-8 days; 7 patients (30%) were treated with non steroidal anti-inflammatory drugs (NSAIDs) only. Eight (35%) patients were treated with both NSAIDs and corticosteroids. Hypothyroidism, with elevated TSH, was observed in 6 (26%) of our patients with positive thyroid antibodies during the first 6 months of follow up. There were no reported cases of recurrent or permanent hypothyroidism in our cohort study. CONCLUSION: Subacute thyroiditis is an uncommon disease that should be considered in the differential diagnosis of acute anterior neck pain, sore throat and fever especially in patients who do not respond to treatment. In the clinical setting, radioiodine uptake can help exclude other diseases, confirm the diagnosis and expedite the initiation of appropriate therapy to relieve symptoms. PMID- 15900375 TI - Frequency of isolated positive sputum cultures among pulmonary tuberculosis patients. AB - OBJECTIVE: The occurrence of isolated positive sputum cultures among pulmonary tuberculosis (PTB) patients during treatment is not widely reported. This study describes the frequency of isolated positive sputum cultures among PTB patients after initiation of chemotherapy. METHODS: Fifty sputum culture positive PTB patients, consisting of 38 (76%) males and 12 (24%) females with a mean age of 34.31 +/- 19.54 (13-75) years, were studied prospectively over a period of 10 months (September 1999 to June 2000) at Sahary Chest Hospital, Riyadh, Kingdom of Saudi Arabia. The majority of patients received standard 4-drug short-course antimicrobial chemotherapy. Weekly sputum smears and cultures were carried out for each patient during the treatment. RESULTS: Isolated positive sputum cultures were encountered in 4 (8%) of the studied patients. Two of them had 2 consecutive positive smears and cultures in their third and fourth week during treatment and were attributed to poor compliance to the treatment. Of the remaining 2 patients, one had single isolated culture in his fourth week, while the fourth patient showed a slight growth in the fifth week of the treatment. Due to the undetermined status of the smear and very few colonies in the culture, these 2 isolated cultures were attributed to the carry-over contamination during the laboratory procedures. The same treatment was continued with strict monitoring of compliance to the treatment and laboratory protocols. All of the 4 patients converted to negative before leaving the hospital. CONCLUSION: Isolated positive sputum cultures might appear (infrequently) during treatment either due to the treatment noncompliance or carry over contamination. However, under either circumstance, the same treatment should be continued along with strict monitoring of treatment compliance and specimen decontamination and related laboratory protocols. PMID- 15900376 TI - Indications, diagnostic yields and complications of transbronchial biopsy over 5 years in the State of Qatar. AB - OBJECTIVES: To review the indications, diagnostic yields and complications of transbronchial biopsy (TBB) in a tertiary hospital in the State of Qatar. METHODS: A retrospective review of our records revealed 1006 adult flexible fibre optic bronchoscopies (FFB) at Hamad General Hospital, State of Qatar between January 1999 and December 2003. A total of 85 (8.4%) TBB were performed, but complete data were available for 71/85 (83.5%), which were reviewed for indications, diagnostic yields and complications. RESULTS: Adequate samples were obtained in 58/71 TBBs (81.7%), while 13/71 TBBs (18.3%) yielded bronchial mucosa. The main indications in 16/71 (22.5%) TBBs for radiographic localized pulmonary disease were to rule out tuberculosis (TB) in 13 cases, and malignancy in 3 cases. Tuberculosis was verified in 3 (23%) of the 13 cases with localized disease. Fifty-five out of 71 (77.5%) TBBs were performed for radiographic diffuse pulmonary disease: 16/55 (29%) for miliary shadows, while 39/55 (70.9%) were carried out for reticular/reticulonodular infiltrates. Histopathology showed granulomatous lesions consistent with TB in 10/16 (62.5%) cases of miliary shadow. In the other pattern of diffuse disease, the histopathological diagnosis were obtained in 25/39 (64%) cases. It showed non-specific pulmonary fibrosis in 13 cases, sarcoidosis in 4 cases, connective tissue disease associated interstitial fibrosis in 4 cases, bronchiolitis obliterans organizing pneumonia (BOOP) in one case, eosinophilic pneumonia in one case, amiodarone toxicity in one case and lymphangitis carcinomatosis in one case. The main complications were minor bleeding <50 cc in 17 cases (23.9%), pneumothorax in 7 cases (9.8%)) and one case had sepsis. CONCLUSION: Our experience substantiates previous reports of the value and safety of transbronchial biopsy in the rapid diagnosis of smear negative miliary TB. In diffuse lung diseases of a non-infectious nature, other than sarcoidosis, lymphangitis carcinomatosis and few other conditions, a pathological diagnosis are much less likely to be reliably made on small pieces of tissue such as those provided by TBB. PMID- 15900377 TI - Mammographic features of isolated tuberculous mastitis. AB - OBJECTIVE: To present the mammography findings in 8 patients with tuberculosis (TB) of the breast, with a review of the literature. METHODS: This study is a retrospective data collection. Each chart with confirmed breast TB based on bacteriology or pathologic findings was analyzed for clinical presentation, gender, nationality, demographic data, prior history of TB, investigation, management, mammographic findings and ultrasound, when available. Mammograms were reviewed by 2 consultant radiologists without knowing the previous diagnosis or the nature of the study. The study was carried out at The State Tuberculosis Registry and Radiology Department, Hamad General Hospital, State of Qatar, from 1990 to 2002. RESULTS: Out of 13 females with TB mastitis, only 8 cases had mammograms preoperatively. The incidence of breast TB in Qatar is rare (1/1000 mammograms per year). Three types of TB mastitis were identified in our study; the nodular (50%), the diffuse (37.5%) of which 77% were limited to one sector of the breast and the sclerosing (12.5%) mastitis. Three patients (43%) were reported as carcinoma. CONCLUSION: Although mammography identified 3 types of TB, it was not helpful in differentiating TB from carcinoma of the breast. However, the careful evaluation of the degree of density and trabecular thickening of the mass in relation to it size might reduce the number of false positive cases of carcinoma diagnosed with mammograms. Biopsy specimen remains the best diagnostic tool in TB mastitis. PMID- 15900378 TI - Lack of effect of N-acetylcysteine treatment to ameliorate the progression of multiple organ failure. AB - OBJECTIVE: To investigate whether prolonged infusion of the oxygen free radical scavenger N-acetylcysteine (NAC) that is commenced immediately after admission to intensive care unit (ICU) could ameliorate the development or progression of multiple organ failure (MOF). METHODS: After receiving ethical committee approval, a prospective randomized, double-blind, placebo controlled study was performed in the Anesthesiology and Reanimation Intensive Care Unit, Hacettepe University Hospital, Ankara, Turkey between December 2002 and May 2003. Twenty six patients were randomized to receive either NAC in 5% dextrose 40 mg/kg/day or the same volume of 5% dextrose both in 4 divided doses. Two patients were withdrawn due to ICU stay <24 hours. Treatment effect on organ function was assessed by the sequential organ failure assessment (SOFA) scores according to physiological parameters of respiratory, hematological, hepatic, cardiovascular, central nervous system (CNS) and renal system scores that were obtained on admission, then daily. Chi-square, Mann Whitney U tests were used for statistical analysis. RESULTS: There was no significant difference between the 2 groups in any of the 5 organ dysfunction parameters, total maximum SOFA, delta SOFA length of intensive care stay, days of mechanical ventilation and mortality. In the NAC treatment group, the maximum SOFA coagulation score was higher than the control group (p<0.05). CONCLUSION: N-acetylcysteine (40 mg/kg/day) that was commenced immediately after admission to ICU did not ameliorate the progression of MOF in this small cohort of patients. We believe routine prophylactic use of low-dose NAC in all critically ill patients does not provide positive protection. PMID- 15900379 TI - Treatment of nosocomial meningitis due to a multidrug resistant Acinetobacter baumannii with intraventricular colistin. AB - We report a case of serious nosocomial meningitis due to a multidrug-resistant Acinetobacter baumannii in a 23-year-old woman who had a posterior fossa craniotomy with upper cervical laminectomy for excision of a meningioma at the level of foramen magnum. Post-operatively, she had neck pain with continuous fever and deterioration in the level of consciousness and convulsions. The CSF was turbid and had neutrophil pleocytosis. A multidrug-resistant Acinetobacter baumannii was isolated from the blood and CSF. The patient failed high doses of imipenem, ciprofloxacin and systemic colistin but responded well to intraventricular injections of colistin 125,000 units twice daily for 3 weeks. No apparent side effects were noticed. We have reviewed other similar cases reported in the literature. PMID- 15900380 TI - Spondylodiscitis developing after epidural catheter use by direct contamination. AB - Spondylodiscitis is a rare but serious complication of epidural catheter use. It is controversial as to which mechanism is responsible for development of this complication. It may develop due to direct contamination or hematogenous spreading. We report a 54-year-old woman who developed spondylodiscitis after epidural catheter was inserted and to whom analgesic and steroid treatment was given for chronic lumbar pain. In this case, each of 2 pathological mechanisms may be responsible. The diagnostic methods, mechanisms and management are discussed. PMID- 15900381 TI - An unusual cause of pelvi-ureteric junction obstruction. AB - Congenital lumbar hernia is rare in the pediatric age group. This report describes an infant with a superior lumbar hernia with herniation of the kidney that resulted in pelvi-ureteric junction obstruction. Surgical repair of the hernia and reduction of the herniated kidney resulted in resolution of the pelvi ureteric junction obstruction. The literature on the subject is also reviewed. PMID- 15900382 TI - High resolution computed tomography appearances of late sequelae of barium aspiration in an asymptomatic young child. AB - Barium aspiration is a well-known complication of upper gastro-intestinal studies. Consequences of aspiration are generally insignificant and leave no permanent changes in the lung parenchyma. However, large quantities of high density barium, if aspirated, lead to silent interstitial changes and fibrosis. High-resolution computed tomography HRCT appearances of lung changes have been demonstrated in adults; few such reports are available in the pediatric literature. We report a case of a child who aspirated barium 3 months before this presentation. The HRCT appearances of barium aspiration are presented with a review of the literature. PMID- 15900383 TI - Severe hypertension secondary to renal artery stenosis and Cushing's syndrome. AB - We present an unusual patient who simultaneously had severe renal artery stenosis RAS and Cushing's syndrome. The case highlights the difficulty of reaching a specific diagnosis of Cushing's syndrome and the possible interaction between Cushing's syndrome and some other concurrent illnesses that this patient had. A 37-year old man presented with severe hypertension HTN and uncontrolled diabetes mellitus DM without clear physical signs of Cushing's syndrome. He was found to have severe osteoporosis, proximal myopathy, several cutaneous warts, tinea versicolor, and chronic viral hepatitis. Captopril-stimulated renal scan and renal artery angiogram revealed severe RAS. Partial balloon dilatation of RAS led to improvement in HTN. Unexpectedly, urine free cortisol 24 hour was found extremely high. Serum adrenocorticotropic hormone ACTH was also elevated and high dose dexamethasone suppression tests were inconclusive. Several imaging studies failed to localize the source of ACTH. Despite normal MRI of the pituitary gland, bilateral inferior petrosal sinus sampling IPSS localized the source of ACTH secretion to the right side of the pituitary gland and right anterior hemihypophysectomy resulted in cure of Cushing's disease, HTN, DM, and tinea versicolor with significant improvement in cutaneous warts, osteoporosis, and chronic hepatitis. In conclusion, RAS and Cushing's syndrome may occur together. Significant hypercortisolemia can occur without clear signs of Cushing's syndrome. Controlling hypercortisolemia is of paramount importance when treating chronic infections in patients with Cushing's syndrome. PMID- 15900384 TI - Hemoglobin D/beta-thalassemia and beta-thalassemia major in a Saudi family. AB - The present report describes the clinical, hematological and molecular characteristics in a family with unique interaction between 3 different mutations discovered during routine workup for bone marrow transplantation. In this report, complete hematological and molecular studies were performed for a large Saudi family. The family consisted of parents and 9 children, which revealed that the father is compound heterozygous for hemoglobin Hb D Punjab/beta-thalassemia, the mother is a carrier for beta-thalassemia and 3 of their children are transfusion dependent beta-thalassemia. Two of the children are compound heterozygous for Hb D Punjab/beta-thalassemia like the father but with different genotype. The other 2 children have Hb D Punjab traits while 2 other children have beta-thalassemia traits. Although, compound heterozygous for Hb D/beta-thalassemia has been well described in the literature, our report emphasizes the importance of careful analysis of the electrophoresis results and the usefulness of molecular studies in premarital screening and other screening hemoglobinopathy programs. PMID- 15900385 TI - Purulent pericarditis due to Staphylococcus aureus in a patient with fever, cough and chest pain. PMID- 15900386 TI - Epidemiology of dermatophyte infection. Comparison of clinical and mycological findings. PMID- 15900387 TI - Assessment of toxoplasma IgG avidity test results in pregnant women. PMID- 15900388 TI - Association between angiographic evidence of coronary artery disease and seropositivity for Helicobacter pylori and Chlamydia pneumoniae infection. PMID- 15900389 TI - Screening for glucose-6-phosphate dehydrogenase deficiency in Behcet's disease. PMID- 15900390 TI - Abruptio placentae following snake bite in a Sudanese woman. PMID- 15900391 TI - Prevalence of smoking and frequency of visits to primary health care clinics. PMID- 15900392 TI - Surgical beds in neonatal intensive care unit. PMID- 15900393 TI - Which local anesthesia should be used in neonatal circumcision in newborns? PMID- 15900394 TI - Scoring systems for diagnosing acute appendicitis. PMID- 15900395 TI - Communities adjust their temperature optima by shifting producer-to-consumer ratio, shown in lichens as models: II. Experimental verification. AB - The community adaptation hypothesis [7] predicts that lichens, simple communities of microorganisms, can adapt to a wide range of thermal regimes by regulating the ratio of primary producers (algae) and consumers (fungi): R(p/c). To test this hypothesis, we determined R(p/c) values by image analysis of cross sections of herbarium specimens of the lichen Cladina rangiferina, which is widely distributed between the Arctic and the tropics. We found that R(p/c) for C. rangiferina increases with summer temperature by more than one order of magnitude, consistent with the hypothesis. To assess the ecological significance of community adaptation (R(p/c) regulation), other adaptive mechanisms (e.g., photobiont substitution, genetic adaptation, and photosynthetic acclimation in North American Cladina spp.) were studied. Laboratory investigations with algae and fungi isolated in culture from live specimens suggested that the role of these mechanisms is relatively minor and cannot account for the high degree of lichen adaptability. PMID- 15900396 TI - Primary intracranial hydatid cyst in the interpeduncular cistern. AB - Cerebral involvement in hydatid disease occurs in 1-4% of cases. There are few documented cases in the literature of intracranial hydatid cysts in unusual locations such as pons and thalamus. In this report, a case of 33-year-old male with primary intracranial hydatid cyst in the interpeduncular cistern is reported. This is the first such case in the literature. PMID- 15900397 TI - Intracranial pressure changes during fluid percussion, controlled cortical impact and weight drop injury in rats. AB - BACKGROUND: In traumatic brain injury research, the fluid percussion injury (FPI) model in the rat is widely used. The injury is graded based on indirect criteria, such as the extracranial pressure wave and/or physiological responses to the injury. We designed this study to investigate if the extracranially monitored pressure in the FPI-device corresponded to the actual intracranial situation. Severe controlled cortical impact (CCI) and severe weight drop injury (WDI) were studied for comparison. METHOD: We tested the correlation between the extra- and intracranial pressures during severe FPI in rat (2.6-2.9 atm), using pressure probes (diameter 0.34 mm) with high frequency (500 Hz) and high pressure range (1 5 atm). The probes were inserted into either of the lateral ventricles in FPI and in the contralateral lateral ventricle in CCI and WDI to compare the ictal pressure pulses between the models. FINDINGS: FPI showed a time lag between the extracranial, intracranial ipsilateral and intracranial contralateral pressure curves respectively, reflecting the different distances between the pressure source and the individual pressure probes. There was a high degree of correlation (r = 0.994, p<0.0001) between the extra- and intracranial pressure pulses, once corrected for the time lag. We found no significant differences between the extracranial and the intracranial peak pressure in either ventricle in FPI. In CCI and WDI the contralateral pressure pulses were significantly smaller than in FPI. CCI resulted in higher pressure peaks than WDI, due to higher impact velocity. CONCLUSIONS: The extracranial pressure pulse appears to be a good estimate of the intraventricular pressure pulse generated during FPI. Severe CCI and WDI generated intraventricular pressure pulses of much lower magnitude than FPI, explaining the lesser degree of brain stem involvement in the former models. PMID- 15900398 TI - Cerebral arteriovenous malformations: when is surgery indicated? PMID- 15900399 TI - Microsurgical results with large vestibular schwannomas with preservation of facial and cochlear nerve function as the primary aim. AB - OBJECTIVE: To evaluate our microsurgical results in dealing with vestibular schwannomas (VS) greater than or equal to 30 mm when preservation of cranial nerve function was considered more important than total tumour removal. METHODS: Sixteen consecutive cases were operated on by the same neurosurgeon according to a prospective protocol using intraoperative neuro-monitoring (IONM) based on electromyographic and brain stem auditory evoked potential recordings. Facial nerve function was evaluated on the House-Brackmann Scale and cochlear nerve function on the Gardner-Robertson Scale. Someone not involved in the clinical management of our patients collected all data. RESULTS: Fifteen patients showed facial nerve (FN) function of House-Brackmann grade (HBG) I or II at one year postoperatively and one kept the HBG IV she had preoperatively. Two patients of four maintained a cochlear nerve function of Gardner-Robertson grade (GRG) II. The tumour excision rates were: total, 68.7%; near total, 6.3%; subtotal, 18.7%, and partial, 6.3%. The average follow-up was 55 months (1-106). Three patients underwent radiotherapy later with growth stabilisation and no additional morbidity. CONCLUSION: When dealing with VS greater than or equal to 30 mm, microsurgery guided by IONM, with a rate of total or near-total tumour excision of about 75%, can retain socially acceptable facial nerve function (HBG I or II) in all cases and serviceable hearing (GRG I or II) in two cases out of four. Maintaining serviceable cranial nerve function should take precedence over total tumour excision. PMID- 15900400 TI - Rerupture of cerebral aneurysms during angiography--a retrospective study of 13 patients with subarachnoid hemorrhage. AB - OBJECTIVE, BACKGROUND: Cerebral angiography, performed within 24 hr of aneurysmal rupture, carries an increased risk of rebleeding. We have investigated the rerupture rate during angiography procedures under deep general anesthesia and the factors that contribute to rebleeding. METHODS: We divided 69 patients who had experienced aneurysmal rerupture into 2 groups. Group I (n = 13) suffered rebleeding during cerebral angiography and group II (n = 56) who rebled at a different time. We assessed the effects on rebleeding of the (1) time between the first insult and angiography, (2) WFNS clinical grade on admission, (3) blood pressure during angiography, (4) age and sex, (5) Fisher classification on admission, (6) aneurysmal site, and (7) Glasgow outcome score (GOS). RESULTS: Factors that had a statistically relevant effect on rebleeding during cerebral angiography (Group I) were the performance of angiography within 3 hr of the initial insult, the admission grade, and the aneurysmal site. Especially, the rerupture events during cerebral angiography were concentrated within 3 hr of the initial insult; the rate was 23.9% when angiograms were obtained within 3 hr of onset. Group I patients manifested a worse clinical grade and middle cerebral artery (MCA) aneurysms were prevalent in this group. However, there was no significant difference between the 2 groups with respect to blood pressure, age, sex, Fisher classification, and GOS. CONCLUSIONS: Cerebral angiography at ultra early timing (within 3 hr of the insult) carries a high risk of aneurysmal rerupture, even if the procedure is performed under deep anesthesia and normotensive blood pressure. Cerebral angiography during that period should be avoided. PMID- 15900401 TI - Magnetic resonance imaging in experimental subarachnoid haemorrhage. AB - BACKGROUND: We developed an MRI protocol to measure cerebrovascular diameter and blood flow velocity, and if we could detect cerebrovascular alterations after SAH and their impact on cerebral ischaemia. METHOD: SAH was induced in 15 Wistar rats by means of the endovascular filament method; 6 other rats served as control. MRI measurements were performed on a 4.7T NMR spectrometer 1 and 48 hours after SAH and 9 days thereafter. Diffusion-weighted and T2-weighted images were acquired to detect cerebral ischaemia. The arterial spin labelling method was used to measure CBF. MR angiography was used to measure vessel diameter and blood flow velocity, from which the arterial blood flow was calculated. FINDINGS: The ischemic lesion volume increased between 1 and 48 hours after SAH from 0.039 to 0.26 ml (P = 0.003). CBF decreased from 53.6 to 39.1 ml/100 g/min. The vessel diameter had narrowed, the blood flow velocity diminished as did the arterial blood flow in most vessels, but only the vasoconstriction in the right proximal ICA reached significance (0.49 mm to 0.43 mm, P = 0.016). Baseline values were restored at day 9. CONCLUSIONS: We showed that it is feasible to detect alterations of in vivo vessel diameter and blood flow velocities and their consequences for brain damage after experimental SAH in the rat. The growth of the infarct volume between day 0 and 2 after SAH and the parallel vasoconstriction suggest that delayed cerebral ischaemia after SAH occurs in rats and that this may be caused by vasoconstriction. PMID- 15900402 TI - Continuous cerebral compliance monitoring in severe head injury: its relationship with intracranial pressure and cerebral perfusion pressure. AB - BACKGROUND: Cerebral compliance expresses the capability to buffer an intracranial volume increase while avoiding a rise in intracranial pressure (ICP). The autoregulatory response to Cerebral Perfusion Pressure (CPP) variation influences cerebral blood volume which is an important determinant of compliance. The direction of compliance change in relation to CPP variation is still under debate. The aim of the study was to investigate the relationship between CPP and compliance in traumatic brain injured (TBI) patients by a new method for continuous monitoring of intracranial compliance as used in neuro-intensive care (NICU). METHOD: Three European NICU's standardised collection of CPP, compliance and ICP data to a joint database. Data were analyzed using an unpaired student t test and a multi-level statistical model. RESULTS: For each variable 108,263 minutes of data were recorded from 21 TBI patients (19 patients GCS/=20 mmHg in 20% and CPP<60 mmHg for 10.7% of the time. Compliance was lower (0.51+/-0.34 ml/mmHg) at ICP>/=20 than at ICP<20 mmHg (0.73+/-0.37 ml/mmHg) (p<0.0001). Compliance was significantly lower at CPP<60 than at CPP>/=60 mmHg: 0.56+/-0.36 and 0.70+/-0.37 ml/mmHg respectively (p<0.0001). The CPP - compliance relationship was different when ICP was above 20 mmHg compared with below 20 mmHg. At ICP<20 mmHg compliance rose as CPP rose. At ICP>/=20 mmHg, the relation curve was convexly shaped. At low CPP, the compliance was between 0.20 and 0.30 ml/mmHg. As the CPP reach 80 mmHg average compliance was 0.55 ml/mmHg., but compliance fell to 0.40 ml/mmHg when CPP was 100 mmHg. CONCLUSIONS: Low CPP levels are confirmed to be detrimental for intracranial compliance. Moreover, when ICP was pathological, indicating unstable intracranial equilibrium, a high CPP level was also associated with a low volume-buffering capacity. PMID- 15900403 TI - Cryptic cerebellopontine angle neuroglial cyst presenting with hemifacial spasm. AB - Hemifacial spasm (HFS) is commonly caused by a vascular loop compressing the Root Exit Zone (REZ) of the facial nerve. We report a case of HFS caused by a vascular loop that was abnormally displaced by a neuroglial cyst not seen in Magnetic Resonance Imaging (MRI). Microvascular decompression (MVD) was planned and the patient underwent a key-hole retromastoid posterior fossa exposure. A cystic lesion was found in the cerebellopontine angle (CPA), located around the seventh and eighth cranial nerves extending from the porous acousticus to the brainstem REZ of the facial nerve. The cyst wall was partially excised revealing the region of the neurovascular conflict. MVD of the facial nerve was performed with immediate postoperative complete resolution of the patient's symptoms. PMID- 15900404 TI - Analysis of saliva for glutathione and metabolically related thiols by liquid chromatography with ultraviolet detection. AB - A method for simultaneous determination of glutathione and its precursors cysteine, cysteinylglycine and homocysteine in saliva is presented. The procedure involves reductive conversion of disulfides to thiols, derivatization to their 2 S-quinolinium derivatives with 2-chloro-1-methylquinolinium tetrafluoroborate and separation and quantitation by reversed-phase ion-pairing high performance liquid chromatography with ultraviolet detection at 355 nm. The calibration performed with saliva samples spiked with thiol disulfides, within the practical concentration ranges, showed linear response of the detector. The method applied to the saliva samples donated by volunteers showed mean concentration (SD, n = 8) of cysteine, cysteinylglycine, glutathione and homocysteine: 26.5 (31.6), 6.05 (5.12), 16.97 (7.68), 3.64 (1.34) nmol/ml respectively. PMID- 15900405 TI - Expression of the apoptotic calcium channel P2X7 in the glandular epithelium. AB - In the current study, expression of the apoptotic calcium channel receptor P2X(7) and prostate-specific antigen (PSA) levels were studied in biopsy cores from 174 patients as well as 20 radical prostatectomy cases. In clinical biopsies, we have previously demonstrated that P2X(1 )and P2X(2) calcium channel receptors are absent from normal prostate epithelium that does not progress to prostate cancer within 5 years. In cases that did progress to prostate cancer however, P2X(1 )and P2X(2) labeling was observed in a stage-specific manner first in the nucleus, then the cytoplasm and finally on the apical epithelium, as prostate cancer developed. These markers were present up to 5 years before cancer was detectable by the usual morphological criteria (Gleason grading) as determined by H and E staining. In the current study, the apoptotic calcium channel receptor P2X(7) yielded similar results to that of P2X(1) and P2X(2). Using radical prostatectomy tissue sections as well as biopsies, these changes in calcium channel metabolism were noted throughout the prostate, indicating a field effect. This finding suggests that the presence of a prostate tumor could be detected without the need for direct sampling of tumor tissue, leading to detection of false negative cases missed by H or E stain. The reliability of PSA levels as a prognostic indicator has been questioned in recent years. In the current study, PSA levels were correlated with the P2X(7) labeling results. All patients who exhibited no P2X(7) labeling had a prostatic serum antigen (PSA) level of <2. Patients who exhibited stage-specific P2X(7) expression, and who later developed obvious prostate cancer as diagnosed by H and E stain, all had a PSA > 2. This finding suggests that increasing PSA may be an accurate indicator of cancer development. PMID- 15900406 TI - The tissue distribution of murine Abcc6 (Mrp6) during embryogenesis indicates that the presence of Abcc6 in elastic tissues is not required for elastic fiber assembly. AB - Mutations in the gene coding for the ABC transporter, ABCC6, in humans cause Pseudoxanthoma elasticum, which is characterized by the deposition of aberrant elastic fibers. To investigate whether the presence of ABCC6 in tissues synthesizing elastin is required for elastin deposition and elastic fiber assembly, we have compared the steady-state levels and tissue distribution of Abcc6 and tropoelastin mRNAs during mouse embryogenesis. Whereas tropoelastin mRNA levels rose during embryogenesis and were the highest in neonatal mice, Abcc6 mRNA levels remained constantly low throughout embryogenesis. In some tissues, both Abcc6 and tropoelastin mRNA were detected. However, Abcc6 mRNA and protein were not detected in neonatal aorta and arteries, which produce large amounts of elastin indicating that the presence of Abcc6 in elastic tissues is not required for elastic fiber assembly. PMID- 15900407 TI - Immunolocalization of fibroblast growth factor-2 (FGF-2) in the developing root and supporting structures of the murine tooth. AB - Epithelio-mesenchymal interactions are active during the development of the root of the tooth and are regulated by a variety of growth factors, such as fibroblast growth factors. FGF-2, 3, 4, and 8 have all been shown to play a role in the development of the crown of the tooth, but less is known about the factors that govern root formation, particularly FGF-2. The aim of this study was thus to elucidate the spatial and temporal expression of FGF-2 in the root of the developing tooth, as this growth factor is believed to be a mediator of epithelio mesenchymal interactions. Parasagittal sections of the maxillary and mandibular arches of post-natal mice were utilized and the roots of the molar teeth were studied. Immunocytochemistry utilizing an antibody to FGF-2 was performed on sections of teeth at various stages of development. Intense immunostaining for FGF-2 was observed in differentiating odontoblasts at the apical end of the tooth and in the furcation zone of the developing root at all the stages examined. FGF 2 localization was also observed in cementoblasts on post-natal days 16, 20 and 24. The pattern of localization of FGF-2 in the developing root suggests that this growth factor may participate in the signaling network associated with root development. PMID- 15900408 TI - Endogenous zinc can be a modulator of glycinergic signaling pathway in the rat retina. AB - Zinc is a modulator of glutamatergic inputs in the hippocampus. In the retina, however, we previously reported that endogenous zinc is present in the non glutamatergic neural processes and earlier electrophysiological studies suggest that zinc is a modulator of inhibitory signaling pathways, which are mediated by glycine and GABA. AII amacrine cells, a subpopulation of glycinergic amacrine cells, are identified by selective immunoreactivity for parvalbumin in the rat retina. In the present study, therefore, we focused on whether zinc is present in AII amacrine cells using silver amplification combined with immunohistochemistry in the rat retina. We also examined whether zinc modulate glycine response in the rat retina by the patch clamp technique. Association of silver precipitates with the parvalbumin-immunoreactive neural processes was observed at the ultrastructural level. We also found that zinc existed in the neural processes which were not parvalbumin-immunoreactive. Glycine-induced responses were augmented when the concentration of Zn(2+) was below 10 microM, but inhibited at Zn(2+) concentrations of 50 microM or more. Our results suggest the notion that zinc in neural processes of retinal neurons modulates the inhibitory signaling pathway, particularly that mediated by glycine receptors in AII amacrine cells. PMID- 15900409 TI - Localization of nitric oxide synthase in rat trigeminal primary afferent neurons using NADPH-diaphorase histochemistry. AB - Nitric oxide (NO) is a ubiquitous gaseous neurotransmitter that has been ascribed to a large number of physiological roles in sensory neurons. It is produced by the enzyme nitric oxide synthase (NOS). To identify the NOS-containing structures of rat trigeminal primary afferent neurons, located in the trigeminal ganglion (TrG) and mesencephalic trigeminal nucleus (MTN), histochemistry to its selective marker nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was applied in this study. In the TrG approximately half of the neuronal population was NADPH-d reactive. Strongly positive were neurons mainly of small-to-medium size. Neuronal profiles of large diameter were less intensely stained. In addition, NADPH-d-positive nerve fibers were dispersed throughout the ganglion. Nitrergic neurons were located in the caudal part and mesencephalic-pontine junction of the MTN. Most of them were large-sized pseudounipolar cells. In a more rostral aspect, the reactive psedounipolar MTN profiles gradually decreased in number and intensity of staining. There, only a fine meshwork of stained thin fibers and perisomatic terminal arborizations, and also some isolated perikarya of NADPH-d stained multipolar MTN neurons, were observed. The predominant NADPH-d localization in smaller in size TrG neurons, which are considered nociceptive, suggests that NO may play a role in the pain transmission in the rat trigeminal afferent pathways. In addition, the wide distribution of NADPH-d activity in large pseudounipolar and certain multipolar MTN neurons provides substantial evidence that NO may also participate in mediating proprioceptive information from the orofacial region. The differential expression patterns of nitrergic fibers in the TrG and MTN suggest that trigeminal sensory information processing is controlled by nitrergic input through different mechanisms. PMID- 15900410 TI - Expression of p57KIP2 in infantile hemangioma. AB - AIM: To compare the expression of p57 as indirect marker of genomic imprinting of CDKN1C in a series of infantile hemangiomas (IH) of patients with and without Beckwith-Wiedemann syndrome. MATERIALS AND METHODS: Cases of mammary, salivary gland, liver (one each), and placental (2 cases) capillary hemangiomas all with histological features akin to IH as well as typical examples of cutaneous (8 cases) IH were analyzed by immunohistochemistry with antibody against p57(KIP2). This protein is the product of CDKN1C an imprinted, maternally expressed gene. The liver hemangioma and both chorioangiomas were from patients with Beckwith Wiedemann syndrome. Positive and negative controls included normal placental tissue and complete hydatidiform mole, respectively. Positive staining was localized to nuclei. RESULTS: Endothelial cells from the skin, breast and salivary gland hemangiomas were p57(KIP2) positive while chorioangiomas and liver IH presenting in patients with Beckwith-Wiedemann syndrome were negative. Controls reacted appropriately. CONCLUSIONS: Endothelial cells of IH not associated with BWS normally express p57(KIP2) while chorioangiomas and IH of the liver associated with BWS do not. These results suggest that the BWS IH may result from dysregulation of the cell cycle. PMID- 15900411 TI - Active participation of apoptosis and mitosis in sublingual gland regeneration of the rat following release from duct ligation. AB - This study was designed to establish how mitotic cell proliferation and apoptotic cell death participate in the regeneration of atrophied rat sublingual glands. To induce atrophy to the sublingual gland of rats, the excretory duct was ligated unilaterally near the hilum, and after 1 week of ligation (day 0) the duct ligation was released to enable gland regeneration. The regenerating glands were examined with routine histology, immunohistochemistry for proliferating cell nuclear antigen (PCNA) as a marker of proliferating cells, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) as a marker of apoptotic cells, and transmission electron microscopy. At day 0, a few acini and many ducts remained in the atrophic sublingual glands, and newly formed immature acini were observed at day 3. Thereafter acinar cells progressively matured and increased in number, although the number of ducts decreased. Many PCNA- and some TUNEL-positive cells were seen in acini and ducts during regeneration. The labeling indices for both cell types were statistically significantly different from that of the control at several time points of the regeneration. Apoptotic and mitotic cells were also confirmed to be present in the experimental sublingual glands by electron microscopy. These observations suggest that apoptosis as well as mitosis of duct and acinar cells actively participate in and play important roles in sublingual gland regeneration. PMID- 15900412 TI - The tidemark of the chondro-osseous junction of the normal human knee joint. AB - The chondro-osseous junction includes the junction between calcified and non calcified cartilage matrices often referred to as the tidemark. A detailed knowledge of the structure, function and pathophysiology of the chondro-osseous junction is essential for an understanding both of the normal elongation of bones and of the pathogenesis of osteoarthrosis. In this study the molecular anatomy of the tidemark was studied using histochemical techniques, including lectin histochemistry, on blocks of normal cartilage from human knee joints. The tidemark stained with H and E, picro-sirius red, toluidine blue, safranin O and methyl green, but not with alcian blue in the presence of magnesium chloride at 0.05 M or above. It stained with only four lectins, those from Datura stramonium, Maclura pomifera, Erythrina crystagalli and Helix pomatia, out of the 19 used. Therefore, it is rich in collagen and contains hyaluronan, but appears to lack the glycosaminoglycans of 'conventional' proteoglycans and it expresses a very limited and distinctive lectin staining glycoprofile, which is probably attributable to specific glycoproteins. In addition, the tidemark had a distinct microanatomical trilaminate appearance. From all of these results it is clear that this part of the chondro-osseous junctional region is chemically more complex and distinctive than has previously been described. PMID- 15900413 TI - Balance between hypertrophic and hypoxic stimulus in caspase-3 activation during rat heart development. AB - During heart development, cell hyperplasia and hypertrophy are the main mechanisms by which cardiac mass grows. Both these processes along with programmed cell death lead to complete growth and function. In addition, since the establishment of cardiac function depends on the relationship between oxygen supply and demand, we investigated some of the molecular mechanisms at the basis of rat myocardial cell response to hypoxic stress at different times of neonatal life. In particular, the role played by hypertrophic and survival factors like NF kB and IAP-1 (Inhibiting Apoptosis Protein) and by death factors ASK-1 (Apoptosis Signal Regulating Kinase), JNK/SAPK (Jun-N-Terminal-Kinase/Stress-Activated Protein Kinase) pathways in regulating caspase-3 expression and activity has been evaluated by immunohistochemical and Western blotting analyses, respectively. Level of phosphorylation of IkBalpha and IAP-1 expression were substantial in 8 day-old hypoxic hearts, suggesting the persistence of NF-kB driven hypertrophic signal along with a rescue attempt against hypoxic stress. In contrast, ASK-1 mediated JNK/SAPK activation, regulating Bcl(2) levels, allows Bax homodimerization and caspase-3 activation in the same experimental conditions. Thus, a regulation carried out by NF-kB and JNK/SAPK pathways on caspase-3 activation at day 8 of neonatal life can be suggested as the main factor for the heart 'adaptive' response to hypoxia. PMID- 15900414 TI - Implications of ethnic group origin for Israeli women's mental health. AB - Israel is an immigrant society comprised of diverse ethnic groups differentiated by variables such as emigration history, health status, educational level, and economic status. The major question addressed in this study is whether differences in women's mental health status are related to ethnic group origin, per se, or rather can be explained by the associated social-demographic strata. The impact of ethnic group origin, social support, health status, education, and economic difficulties was assessed on a set of mental health outcomes. The analysis was carried out in a random sample of 522 women residing in the Negev region of Israel-a culturally diverse area. The findings indicate that social support, economic status, health status, and ethnic group origin all contribute to shaping women's mental health status. PMID- 15900415 TI - Medical complaints among Iraqi American refugees with mental disorders. AB - The Gulf War in 1991 resulted in an influx of refugees from Iraq to the United States and to other regions of the world. The purpose of this study was to describe the self-reported medical complaints of Iraqi American refugees who were seeking mental health services in southeastern Michigan. We anticipated that the frequency and pattern of medical symptoms would differ from that reported in the literature on United States Gulf War veterans or other Arab Americans who immigrated to the U.S. in the 1990s. Potential reasons for such differences include indirect effects, such as neglect of general health prior to and during the war, or direct effects, such as the impact of environmental changes from the war itself. As part of a larger study on the health of refugees from Iraq, self reported medical conditions and symptoms were analyzed in a sample of 116 adult Iraqi immigrants (46 male, 70 female) who were seeking or already receiving outpatient mental health services (n = 87) or treatment in a partial hospitalization program (n = 29). Measures were translated into Arabic and administered in an interview format by one of two bilingual mental health workers. The results were consistent with other studies on refugees in which the number of medical complaints reported was relatively high. Discussion centers on the importance of addressing the specific medical needs of refugees in general, and of the Iraqi refugees in particular, and on how they may be better served within our primary health care systems. PMID- 15900416 TI - Making sense of illness: late-in-life migration as point of departure for elderly Iranian immigrants' explanatory models of illness. AB - This article is based on data gathered through 60 qualitative interviews conducted within the realm of three research projects that have used "culture appropriate lenses" to study the postmigration situation of late-in-life Iranian immigrants to Sweden. The findings gathered through these studies were interpreted against the backdrop that culturally appropriate nursing theories provide. This meant that it was, at times, these elders' backgrounds as cultural "others" that were implicitly used to make sense of the various issues that were brought to the fore by these studies. The particular issue with which this article is concerned is the "unusualness" of these elders' explanatory models of illness. Inspired by the concept "definition of situation" in the symbolic interactionist perspective and by the feeling that this perspective might bring about a different interpretation of the original findings regarding their understandings of illness and disease, we set out to conduct a secondary analysis of these elders' explanatory models of illness. The findings presented in this article will show how the elderly Iranian immigrants interviewed in these three studies utilize the process of "late in life migration" as a point of reference for their understandings of what has caused the illnesses from which they suffered. Hereby we will suggest that the "unusualness" of their explanatory models of illness might be best understood if we focus on what they shared as immigrants (i.e., the fact that the process of late-in-life migration has made their culture obsolete) as opposed to what they shared as Iranians (i.e., their culture of origin). PMID- 15900417 TI - Asthma prevalence and severity in Arab American communities in the Detroit area, Michigan. AB - Immigrant populations provide a unique intersection of cultural and environmental risk factors implicated in asthma etiology. This study focuses on asthma prevalence and severity in 600 Arab American households in metro Detroit, the largest immigrant reception zone for Arab Americans in North America. The survey method introduced a number of novel features: (a) a ranking scheme for the key environmental risk factors for asthma was used to derive an aggregated environmental risk index (ERI) for each household, and (b) an aggregate measure of asthma severity based on symptom frequency and intensity. Environmental risk factors and surrogates for socioeconomic status (SES) were found to be stronger predictors of asthma prevalence than asthma severity, while demographic variables such as English fluency and birth in the United States were better predictors of asthma severity than asthma prevalence. These results suggest that SES variables may be more reflective of environmental exposures in communities involved in this study, while English fluency and birth in the United States may be linked to health care access and utilization behavior that can influence the asthma management. We also found a significant relationship between asthma prevalence and degree of acculturation. Asthma prevalence was highest among moderately acculturated immigrants compared with new immigrants and those who were well acculturated, suggesting that among Arab Americans in the Detroit area, risk factors associated with new immigrant status are replaced by "western" risk factors as the population becomes more acculturated. PMID- 15900418 TI - Measuring acculturation among male Arab immigrants in the United States: an exploratory study. AB - Given the absence of empirical measures that assess acculturation patterns among male Arab immigrants, a new acculturation instrument was developed and evaluated. One hundred and fifteen adult male Arab immigrants were administered the Male Arab Acculturation Scale (MAAS), and psychometrically established measures of ethnic identity and self-esteem. Satisfactory reliability is reported for the two acculturation scales, separation/assimilation and integration/marginalization. Ethnic identity, self-construal, personal self-esteem, and collective self-esteem appear to be differentially associated with acculturation patterns among male Arab immigrants. PMID- 15900419 TI - Refugees' perceptions of healthy behaviors. AB - This descriptive study explored refugees' knowledge and perceptions of nutrition, physical activity and smoking behaviors using the Health Promotion and Transtheoretical Models. A one-time interview used both closed- and open-ended questions. The sample included 31 adults from Bosnia, Iran, and Cuba. Refugees had some knowledge of a healthy diet and physical activity, and were aware of both benefits and barriers for health behaviors. They had a realistic perception of their weight (55% overweight), and none thought obesity was a positive characteristic. Changes in diet, physical activity and smoking since arrival in the US have been positive for some and negative for others. For all categories discussed, refugees were in the pre-contemplation stage of change. There is a need to study refugees' health behaviors over time after arrival. Health behavior interventions must be specific to ethnicity in order to accurately document progress and to be culturally appropriate. PMID- 15900420 TI - "It's like playing with your destiny": Bosnian immigrants' views of advance directives and end-of-life decision-making. AB - Patient autonomy is a primary value in US health care. It is assumed that patients want to be fully and directly informed about serious health conditions and want to engage in advance planning about medical care at the end-of-life. Written advance directives and proxy decision-makers are vehicles to promote autonomy when patients are no longer able to represent their wishes. Cross cultural studies have raised questions about the universal acceptance of these health care values among all ethnicities. In the current investigation, Bosnian immigrants were interviewed about their views of physician-patient communication, advance directives, and locus of decision-making in serious illness. Many of the respondents indicated that they did not want to be directly informed of a serious illness. There was an expressed preference for physician- or family-based health care decisions. Advance directives and formally appointed proxies were typically seen as unnecessary and inconsistent with many respondents' personal values. The findings suggest that the value of individual autonomy and control over the health care decisions may not be applicable to cultures with a collectivist orientation. PMID- 15900421 TI - Implications of black immigrant health for U.S. racial disparities in health. AB - This paper contributes to a growing understanding of U.S. black-white health disparities by using national-level data to disaggregate the health status of black Americans into the following subgroups: U.S.-born blacks, black immigrants from Africa, black immigrants from the West Indies, and black immigrants from Europe. Using new data on the 2000 and 2001 National Health Interview Surveys (NHIS), the authors compare the status of U.S.- and foreign-born blacks to that of U.S.-born whites on three measures of health. The analysis finds that U.S. born and European-born blacks have worse self-rated health, higher odds of activity limitation, and higher odds of limitation due to hypertension compared to U.S.-born whites. In contrast, African-born blacks have better health than U.S.-born whites on all three measures, while West Indian-born blacks have poorer self-rated health and higher odds of limitation due to hypertension but lower odds of activity limitation. These findings suggest that grouping together foreign-born blacks misses important variations within this population. Rather than being uniform, the black immigrant health advantage varies by region of birth and by health status measure. The authors conclude by exploring the implications of these findings for researchers, health professionals, and public policy. PMID- 15900422 TI - Stress, coping, and health: a comparison of Mexican immigrants, Mexican Americans, and non-Hispanic whites. AB - Mexican immigrants, Mexican-Americans, and non-Hispanic white Americans all face different stressors. Stress-coping strategies may vary for each group as well. We compared relationships among perceived stress, stress-coping strategies, and health-related quality of life (HRQL) in a rural sample of Mexican citizens living in the United States, Mexican-Americans, and non-Hispanic whites. Health related quality of life and stress-coping styles varied among the three groups. Mexican citizens reported significantly better physical functioning than did non Hispanic whites or Mexican-Americans. Mexican-Americans reported significantly better mental health functioning than did non-Hispanic whites or Mexican citizens. Mexican citizens were more likely to use positive reframing, denial, and religion, and less likely to use substance abuse and self-distraction, as stress-coping strategies. Stress-coping style may be a potentially modifiable predictor of physical and mental HRQL, and may account for part of the Hispanic health paradox. PMID- 15900423 TI - Hypoxic versus normoxic external-beam irradiation of cervical carcinoma combined with californium-252 neutron brachytherapy. Comparative treatment results of a 5 year randomized study. AB - PURPOSE: The article focuses on the treatment and protective effects of hypoxyradiotherapy during external-beam irradiation of cervical carcinoma, including paraaortic lymph nodes, combining radiotherapy with californium-252 ((252)Cf) neutron brachytherapy. An analysis of treatment results, early and late side effects and complications is presented. PATIENTS AND METHODS: From January 1989 to May 1997, 307 women with stage IIb and IIIb cervical carcinoma, treated with (252)Cf neutron brachytherapy, were randomly divided into two groups and treated with external-beam irradiation to the paraaortic lymph nodes as follows: 155 patients (59 with stage IIb, 96 with stage IIIb) were treated by external beam irradiation administered as a 60-Gy dose applied under conditions of acute hypoxia; 77 patients (30 with stage IIb and 47 with stage IIIb) received extended field irradiation up to L4 and 78 patients (29 with stage IIb and 49 with stage IIIb) up to T12. 152 patients (58 with stage IIb, 94 with stage IIIb) were treated by external-beam irradiation administered as a 40-Gy dose applied under normal oxygenation conditions. 73 patients (29 with stage IIb and 44 with stage IIIb) received extended-field irradiaton up to L4 and 79 patients (29 with stage IIb and 50 with stage IIIb) up to T12. The same 56 Gy-equivalent (eq) doses at point A and 19 Gy-eq doses at point B were applied intracavitarily in both groups. The total radiation doses at points A and B were 99 and 79 Gy-eq, respectively, for patients treated with external-beam irradiation to 60 Gy under conditions of acute hypoxia. For patients treated with external-beam irradiation to 40 Gy under normal oxygenation conditions, the doses at points A and B were 85 and 59 Gy-eq, respectively. RESULTS: The 5-year overall survival rate for all patients (stages IIb and IIIb) was 7.0% better for patients treated in acute hypoxia than for patients treated under normal oxygenation conditions (78.7% vs. 71.7% [p < 0.16]). The 5-year metastases-free survival rate was better by 11.7% for stage IIIb patients in the hypoxyradiotherapy group with extended field up to T12 as compared to patients with extended field up to L4 (97.4% vs. 85.7% [p < 0.05]). Comparison of metastases-free survival rate of stage IIIb patients after external-beam irradiation with extended field up to T12 in hypoxic condition versus normoxic condition showed a 12% better result for patients in hypoxic condition (97.4% vs. 85.4% [p < 0.04]). Occurrences of symptomatic radiation induced reactions during or shortly after irradiation were more frequently observed in patients treated with a lower dose under normoxic conditions. During the period of 6-12 years after treatment there were no changes in the frequencies of occurrences of late effects and complications. CONCLUSION: The importance of the protective effects of hypoxyradiotherapy for dose escalation in external-beam irradiation of cervical carcinoma, including paraaortic lymph nodes, with regard to an improvement of the cure rates of metastases in paraaortic lymph nodes has been confirmed. PMID- 15900424 TI - [Prognostic factors after surgery and adjuvant radiotherapy in cervical cancers. A retrospective analysis of 298 patients after surgery and adjuvant radiotherapy]. AB - PURPOSE: To determine prognostic factors in patients with cervical cancer treated with surgery followed by radiotherapy. PATIENTS AND METHODS: In a retrospective analysis, patients treated at the Department of Radiotherapy, University of Halle Wittenberg, Germany, from 1980 through 1993 were evaluated for local control, survival and treatment sequelae with special emphasis on prognostic factors. 298 patients (age 23-81 years, median 46 years) with cervical cancers were treated with surgery followed by radiotherapy to a minimum of 50 Gy. 250 patients received radiotherapy after radical hysterectomy with pelvic lymphadenectomy and 48 patients after hysterectomy. Most of the cancers were squamous cell carcinomas (81%) with few adenocarcinomas (12.4%). Grading was G1/G2 in 28.9% of the patients and G3/G4 in 39.6%. RESULTS: The 5-year overall survival was 68.8% for the whole group, 70.5% for squamous cell carcinomas (n = 241), 64.4% for adenocarcinomas (n = 37; not significant), 80% for G1/G2 tumors (n = 86), and 69% for G3/G4 carcinomas (n = 118; p = 0.074). Survival dependent on tumor size and positive nodes was 76.2% (n = 94) for pT1N0, 65.8% for pT1N+ (n = 31), 68.2% for pT2N0 (n = 62), and 41.4% for pT2N+ (n = 35). An impact of grading on 5-year survival was only found in patients with positive nodes: G1/G2N0 84% (n = 62) versus G1/G2N+ 64.8% (n = 23); G3/G4N0 79.6% (n = 71) versus G3/G4N+ 39.9% (n = 31). The number of positive nodes was the strongest prognostic factor. Multivariate analysis (Cox regression) established a risk factor of 3.06 (p < 0.01) for positive nodes, of 1.7 for grading G1/G2 versus G3/G4 (p = 0.087), and of 1.3 for tumor size (pT1 vs. pT2; p = 0.079). There were no differences in the clinical outcome between patients in stage pT1 with or without lymphadenectomy (75% [n = 130] vs. 79% [n = 39]). CONCLUSION: In patients with cervical cancers treated with surgery and radiotherapy, positive nodes were an independent prognostic factor for local control and survival and should be an indication for additional chemotherapy. A value of lymphadenectomy for survival in this group of irradiated patients could not be established. PMID- 15900425 TI - Dose-response relations for anal sphincter regarding fecal leakage and blood or phlegm in stools after radiotherapy for prostate cancer. Radiobiological study of 65 consecutive patients. AB - BACKGROUND: The estimation of the parameters that describe the dose-response relations of anal sphincter regarding the clinical endpoints of fecal leakage and blood or phlegm in stools is important in the optimization of prostate cancer radiotherapy. Also, the validity of the relative seriality model for this clinical case needs to be examined by associating the clinical follow-up results with the predicted complication rates. PATIENTS AND METHODS: In this study, 65 patients who received radiation therapy for clinically localized prostate adenocarcinoma are analyzed. The clinical treatment outcome and the three dimensional dose distribution delivered to anal sphincter were available for each patient. A questionnaire was used for assessing the clinical bowel and urinary symptoms. A maximum likelihood fitting was performed to calculate the best estimates of the parameters used by the relative seriality model. The clinical utilization of the calculated parameters in predicting anal sphincter complication probabilities was illustrated by applying the best estimate of the parameters to a subset of the patient population. RESULTS: The estimated values of the parameters for the two clinical endpoints are D (50) = 70.2 Gy, gamma = 1.22, s = 0.35 for fecal leakage and D (50) = 74.0 Gy, gamma = 0.75, s approximately 0 for blood or phlegm in stools. The standard deviations of the parameters were also calculated together with the confidence intervals of the dose-response curves. The analysis proved that the treatment outcome pattern of the patient material can suitably be reproduced by the relative seriality model (probability of finding a worse fit = 60.2%, the area under the receiver operating characteristic curve = 0.72 and 0.69 and chi(2)-test = 0.97 and 0.86, respectively). CONCLUSION: Fecal leakage is characterized by a medium relative seriality whereas blood or phlegm in stools was found to have strong volume dependence (low relative seriality). Diminishing the biologically effective uniform dose to anal sphincter < 40-45 Gy may significantly reduce the risk of fecal leakage or blood or phlegm in stools for patients irradiated for prostate cancer. PMID- 15900426 TI - Is grading of breast fibrosis with mammography feasible? AB - PURPOSE: To establish a grading system for mammographic fibrosis and correlate it with clinical fibrosis. PATIENTS AND METHODS: Analogous to the LENT/SOMA scale a four-tiered scoring scale of breast fibrosis in mammography (G0 = absent, G1 = barely increased density, G2 = definitely increased density to G3 = very marked density) was established by two observers in a group of 16 patients. Reference mammograms were selected. Independently and blinded to clinical results, three observers scored the fibrosis in mammograms of further 31 patients examined by one radiation oncologist in a cross-sectional follow-up study. Pretreatment parenchyma density was judged according to the American College of Radiology (ACR). Interobserver correlation of mammography scoring as well as correlation of mammography and clinical findings were calculated with Cohen's weighted kappa. All patients had breast-conserving surgery and axillary resection for breast carcinoma T1-2N0-1. The breast was irradiated to a median reference dose of 55 Gy (range 50-60 Gy) with 2 Gy five times weekly or 2.5 Gy four times weekly. Two patients received chemotherapy, 14 patients tamoxifen. Median age was 55 years, median follow-up 8 years (4-15 years). RESULTS: 14 of 31 patients had clinical fibrosis, twelve G1 and two G2. In mammography, mild fibrosis (G1) was seen in 12/12/18 patients (observer 1/2/3) and moderate fibrosis (G2) in 9/10/2 patients. Interobserver correlation for observers 1 and 2 who had developed the score was fair (Cohen's weighted kappa 0.64, 95% confidence interval 0.4-0.88). However, it was weak for observer 3 (0.36 and 0.42, respectively) who relied on reference mammograms only. Independent interobserver correlation of pretreatment breast density was good for all observers (Cohen's weighted kappa 0.73-0.8). The correlation of fibrosis by mammography and palpation was weak (Cohen's weighted kappa 0.32-0.42). CONCLUSION: Grading fibrosis as depicted by mammography is possible, especially if observers prepare by jointly analyzing a training group. It may be useful to study treatment effects, e. g., of fractionation or drugs, because retrospective and repeated analysis is possible. The correlation of mammography with clinical grading should be further evaluated with more objective clinical reference tools. PMID- 15900427 TI - Irradiation causes biphasic neutrophilic granulocyte phagocytic function. AB - BACKGROUND AND PURPOSE: The anti-inflammatory effect of low-dose radiotherapy is clinically well described. Nevertheless, until now neither the optimal dose nor the background of tissue reactions have been defined. The current study examines the influence of low radiation doses on neutrophilic granulocyte function, which could be helpful in finding the optimal dose for either stimulation or suppression of anti-inflammatory activity. MATERIAL AND METHODS: Lymphoprep density gradient-purified neutrophilic granulocytes of three voluntary, healthy donors were used for all experiments. Granulocytes were incubated 48 h in RPMI 1640 and irradiated with single doses of 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 6.0, and 12 Gy using a (137)Cs IBL 437L irradiator. Their function was assessed by measuring granulocytic release of reactive oxygen species (ROS) with luminol enhanced chemiluminescence after stimulation with phorbol myristate acetate (PMA). RESULTS: Relative changes of ROS release (ROS release before stimulation was set to 100%) increased after stimulation with PMA (mean +/- standard deviation [SD]): 0 Gy: 147.6% +/- 60%; 0.5 Gy: 153.6% +/- 70%; 1.0 Gy: 164.9% +/- 63%; 1.5 Gy: 177.8% +/- 66%; 2.0 Gy: 162.5% +/- 57%; 2.5 Gy: 156.2% +/- 60%; 3.0 Gy: 159.2% +/- 60%; 3.5 Gy: 126.9% +/- 55%; 4.0 Gy: 137.9% +/- 71%; 6.0 Gy: 148.3% +/- 65%; 12.0 Gy: 156.1% +/- 52%. The relative ROS release showed a significant increase at 1.5 Gy (p < 0.001) after PMA stimulation and a significant decrease of ROS release at 3.5 Gy (p < 0.005) and less markedly at 4.0 Gy (p < 0.05). 6.0 and 12.0 Gy showed a significant (p < 0.05) increase again. CONCLUSION: This ex vivo in vitro study on native human neutrophilic granulocytes shows an increase at 1.5 Gy and a significant decrease of granulocyte function at 3.5 and 4.0 Gy, as it was described for different other phenomena in low-dose radiotherapy. These results may provide a further explanation for the local anti-inflammatory effect of low-dose ionizing irradiation. PMID- 15900429 TI - Does the imaging method have an influence on the measured tumor height in ruthenium plaque therapy of uveal melanoma? AB - BACKGROUND AND PURPOSE: For ruthenium plaque therapy of uveal melanoma the precise knowledge of the tumor height is necessary. The purpose of this analysis is to examine systematic differences between ultrasound (US), computed tomography (CT), and magnetic resonance imaging (MRI) measurements of tumor height in uveal melanoma. MATERIAL AND METHODS: From 1993 to 2000, 208 patients with uveal melanoma were treated with ruthenium therapy. US was done in 190/208 patients, CT in 147/208 patients, and MRI in 81/208 patients. The measurements from the imaging modalities were compared to each other. Separate comparisons were carried out for tumor size and localization and between two-dimensional (2-D) and three dimensional (3-D) measurements in reconstructed planes. RESULTS: In direct comparison, CT measurements (median: 4.5 mm; range: 1.6-10.5 mm) were nearly identical to MRI measurements (median: 4.5 mm; range: 2.0-11.4 mm), while US yielded the highest values (median: 5.2 mm; range: 2.5-13.4 mm). Linear regression yielded the following values: CT versus US (R(2) = 0.88, correlation coefficient = 1.04), MRI versus US (R(2) = 0.79, correlation coefficient = 0.92), MRI versus CT (R(2) = 0.84, correlation coefficient = 0.90), and "2-D" versus reconstructed "3-D" (R2 = 0.93, correlation coefficient = 0.98). CONCLUSION: Differences between the measurements from all three modalities were generally acceptable, except for small tumors (< or =4 mm; limited spatial resolution of CT/MRI) and temporal or peripheral lesions (angular distortion in US). In special anatomic situations, CT/MRI measurements can also result in distortions. "3-D" measurements can sometimes help in these situations. PMID- 15900430 TI - Individual radiosensitivity does not correlate with radiation-induced apoptosis in lymphoblastoid cell lines or CD3+ lymphocytes. AB - BACKGROUND AND PURPOSE: Spontaneous and radiation-induced apoptosis of lymphoblastoid cell lines (LCLs) derived from healthy donors, cancer patients and donors with radiosensitivity syndromes as well as CD(3+) lymphocytes from patients with > or = grade 3 late toxicity were investigated as a possible marker for the detection of individual radiosensitivity. These investigations are based on the hypothesis that hypersensitive patients have reduced levels of apoptosis after in vitro irradiation as a result of a defect in the signaling pathway. MATERIAL AND METHODS: Epstein-Barr virus-(EBV-)transformed LCLs derived from five healthy donors, seven patients with heterozygous or homozygous genotype for ataxia-telangiectasia or Nijmegen breakage syndrome and five patients with > or = grade 3 late toxicity (RTOG) were investigated. In addition, CD(3+) lymphocytes from 21 healthy individuals and 18 cancer patients including five patients with a proven cellular hypersensitivity to radiation were analyzed. Cells were irradiated in vitro with a dose of 2 and 5 Gy and were incubated for 48 h. Apoptotic rates were measured by the TUNEL assay followed by customized image analysis. RESULTS: Four out of seven radiosensitivity syndrome patients were identified to have an increased cellular radiosensitivity as determined by reduced apoptotic rates after irradiation of their respective LCLs. Comparatively, only two of the five hypersensitive cancer patients were clearly identified by reduced apoptotic rates. Spontaneous apoptotic rates were very homogeneous among all 39 samples from controls and patients, while lymphocytes of all cancer patients showed significantly lower radiation-induced rates. CONCLUSION: Only a subgroup of hypersensitive patients may be identified by reduction of radiation-induced apoptotic rate. It is concluded that the hypothesis according to which hypersensitive cells have reduced levels of apoptosis is only conditionally true. The authors suggest that this assay can be used in combination with additional tests evaluating DNA double-strand break repair, cell-cycle control and chromosomal aberrations for the evaluation for individual hypersensitivity. PMID- 15900431 TI - Radiosurgery of small skull-base lesions. No advantage for intensity-modulated stereotactic radiosurgery versus conformal arc technique. AB - BACKGROUND AND PURPOSE: Intensity-modulated stereotactic radiotherapy (IMSRT) has shown the ability to conform the dose to concavities and to better avoid critical organs for large tumors. Given the availability of an electronically driven micro multileaf collimator, both intensity-modulated stereotactic radiosurgery (IMSRS) and dynamic conformal arc (DCA) technique (DCA) can be performed at the Novalis Shaped Beam Surgery Center, University of Erlangen-Nuremberg, Germany, since 12/2002. This study evaluates both techniques in small skull-base tumors treated with radiosurgery. MATERIAL AND METHODS: Between 12/2002 and 04/2004, a total of 109 radiosurgical procedures were performed in 77 patients, equally distributed between patients with acoustic neuroma (AN), pituitary adenoma (PA) and meningeoma (M). Six index patients (n = 2 AN, n = 1 PA, n = 3 M) routinely planned for dynamic arc stereotactic radiosurgery were replanned using the IMSRS approach (BrainScan, BrainLAB, Heimstetten, Germany). The RTOG radiosurgery quality assurance guidelines, isodose volumes, doses to organs at risk (OAR), and dose delivery criteria were compared. RESULTS: DCA was superior to IMSRS for homogeneity and coverage. IMSRS could keep the high-dose-irradiated volumes (90% isodose volume) lower than DCA in the PA and AN with very small volumes, but all other lower dose volumes were larger for IMSRS. Dose maxima to OAR were higher for IMSRS. Treatment delivery time for IMSRS would clearly exceed treatment time for DCA by a factor of 2-3. The integral absorbed dose to the brain was much higher in the IMSRS than in the DCA approach (factor 2-3). CONCLUSION: RTOG radiosurgery guidelines were best met by the DCA rather than IMSRS approach for the treatment of small skull-base lesions. The IMSRS approach will increase the time for planning, dose delivery and integral dose to the brain. Thus, IMSRT techniques are recommended for fractionated stereotactic radiotherapy to larger volumes rather than for radiosurgery in small skull-base lesions. PMID- 15900432 TI - [Inpatient rehabilitation of chronic dermatoses illustrated by atopic dermatitis]. AB - Atopic dermatitis is defined as a chronically relapsing skin disease resulting from complex interactions between genetic and environmental factors. It usually occurs during early childhood and shows typical clinical manifestations, depending on the patient's age. In cases of chronic atopic dermatitis, negative effects on professional and social activities and participation have to be expected. To counteract or overcome these threatening impairments in the different facets of life, prescribing inpatient rehabilitative measures should be considered early. Dermatological rehabilitation according to guidelines guarantees an interdisciplinary and multimodal treatment of atopic dermatitis. PMID- 15900433 TI - [Bilateral testicular gunshot injuries]. AB - Gunshot injuries to the testicles are rare and usually result in testicular atrophy. In the case of severe bilateral testicular injuries, this could cause not only infertility but also the need for lifetime testosterone-substitution. We report an 18-year-old patient with bilateral testicular gunshot injury. During the surgical exploration an orchiectomy of the complete ruptured left testicle was necessary. Debridement of the damaged tissue and a partial orchiectomy was performed on the right side. After the operation, the patient developed an incretory hypogonadism and oligozoospermia. During follow-up, an improvement in the sperm count and of the hormonal status occurred. These finally reached normal levels. After genital traumata, immediate surgical exploration should be performed. Based on the above results, the patient benefits from conservative debridement and primary repair of the injured testicle, if possible. An improvement in hormonal status and sperm parameters after testicular injury and consecutive testicular malfunction can occur. Regeneration of the testicular tissue seems possible. PMID- 15900434 TI - Speech recognition implementation in radiology. AB - Continuous speech recognition (SR) is an emerging technology that allows direct digital transcription of dictated radiology reports. The SR systems are being widely deployed in the radiology community. This is a review of technical and practical issues that should be considered when implementing an SR system. PMID- 15900435 TI - Atypical MRI appearance of desmoplastic infantile ganglioglioma. AB - We report the atypical MRI features and histopathological findings of a desmoplastic infantile ganglioglioma in an 8-year-old girl. The mass was predominantly solid with a large, solid, non-enhancing exophytic component. The adjacent brain showed cortical necrosis and white-matter gliosis, suggesting earlier hypoxia. PMID- 15900436 TI - Clinical and radiological outcome of hydroxyapatite-coated femoral stem in revision hip arthroplasty. AB - We used a proximally hydroxyapatite-coated femoral stem in revision arthroplasty of 48 cases with aseptic loosening and Paprosky defect class 1 or 2. We reviewed the outcome after 6.1 (4--9.3) years. The clinical outcome was good, with a mean postoperative HHS of 90 (51--100) points. There were five reoperations all on the acetabular side and none for the femoral stem. At follow-up, we observed cancellous sclerosis radiographically in 19 cases-especially in non-tightly fitted stems and mainly in Gruen zones 2 and 6. In 13 cases, cortical thickening was seen, mainly in Gruen zones 3 and 5 and especially in tightly fitted stems. These bony changes were significant and not related to any clinical parameter. They started to appear from 6 months onward, with increasing frequency with longer follow-up. We find that the standard Mallory-Head hydroxyapatite-coated femoral stem is suitable for revision in cases with lower-class femoral defects. PMID- 15900437 TI - Orthopaedic applications of gene therapy. AB - Gene therapy presents a novel approach to biological treatment. Several orthopaedic diseases can cause changes in biological signalling at the tissue level that potentially can be repaired or modified by inserting genes into the cells or tissues to modulate gene expression. Impaired bone healing, need for extensive bone formation, cartilage repair and metabolic bone diseases are all conditions where alterations of the signalling peptides involved may provide cure or improvement. In orthopaedic oncology, gene therapy may achieve induction of tumour necrosis and increased tumour sensitivity to chemotherapy. In the last decade, extensive improvements have been made to optimise gene therapy and have been tested on several orthopaedic conditions. How far this development has come in orthopaedics is highlighted in this paper. PMID- 15900438 TI - Desmoid tumours of the extremities and trunk: a review of the literature. AB - Desmoid tumours are rare neoplasms that display local aggressiveness but no propensity to metastasise. They are mainly localized in the abdominal wall, the bowel, and the mesentery or in extra-abdominal sites such as the trunk and the extremities. Surgical resection is the main treatment modality in extremities and trunk, with the optional combination of radiotherapy and/or chemotherapy. However, these tumours have a high propensity for recurrent growth. PMID- 15900439 TI - Feedforward-feedback control of dissolved oxygen concentration in a predenitrification system. AB - As the largest single energy-consuming component in most biological wastewater treatment systems, aeration control is of great interest from the point of view of saving energy and improving wastewater treatment plant efficiency. In this paper, three different strategies, including conventional constant dissolved oxygen (DO) set-point control, cascade DO set-point control, and feedforward feedback DO set-point control were evaluated using the denitrification layout of the IWA simulation benchmark. Simulation studies showed that the feedforward feedback DO set-point control strategy was better than the other control strategies at meeting the effluent standards and reducing operational costs. The control strategy works primarily by feedforward control based on an ammonium sensor located at the head of the aerobic process. It has an important advantage over effluent measurements in that there is no (or only a very short) time delay for information; feedforward control was combined with slow feedback control to compensate for model approximations. The feedforward-feedback DO control was implemented in a lab-scale wastewater treatment plant for a period of 60 days. Compared to operation with constant DO concentration, the required airflow could be reduced by up to 8-15% by employing the feedforward-feedback DO-control strategy, and the effluent ammonia concentration could be reduced by up to 15 25%. This control strategy can be expected to be accepted by the operating personnel in wastewater treatment plants. PMID- 15900440 TI - Contributing factors affecting the prognosis of surgical outcome for thoracic OLF (S.-U. Kuh et al.). PMID- 15900441 TI - The 1.13-A structure of iron-free cytochrome c peroxidase. AB - The iron-free cytochrome c peroxidase (CCP) crystal structure has been determined to 1.13 A and compared with the 1.2-A ferric-CCP structure. Quite unexpectedly, removal of the iron has no effect on porphyrin geometry and distortion, indicating that protein-porphyrin interactions and not iron coordination or formation of the axial His-Fe bond determines porphyrin conformation. However, there are changes in solvent structure in the distal pocket, which lead to changes in the distal His52 acid-base catalyst. The observed ability of His52 to move in response to small changes in solvent structure is very likely important for its role as a catalyst in assisting in the heterolytic fission of the peroxide O-O bond. PMID- 15900442 TI - The development of the DIGE system: 2D fluorescence difference gel analysis technology. AB - Two-dimensional (2D) gel electrophoresis is a powerful technique enabling simultaneous visualization of relatively large portions of the proteome. However, the well documented issues of variation and lack of sensitivity and quantitative capabilities of existing labeling reagents, has limited the use of this technique as a quantitative tool. Two-dimensional difference gel electrophoresis (2D DIGE) builds on this technique by adding a highly accurate quantitative dimension. 2D DIGE enables multiple protein extracts to be separated on the same 2D gel. This is made possible by labeling of each extract using spectrally resolvable, size and charge-matched fluorescent dyes known as CyDye DIGE fluors. 2D DIGE involves use of a reference sample, known as an internal standard, which comprises equal amounts of all biological samples in the experiment. Including the internal standard on each gel in the experiment with the individual biological samples means that the abundance of each protein spot on a gel can be measured relative (i.e. as a ratio) to its corresponding spot in the internal standard present on the same gel. Ettan DIGE is the system of technologies that has been optimized to fully benefit from the advantages provided by 2D DIGE. PMID- 15900443 TI - Quality assurance challenge 1. PMID- 15900444 TI - Solution to informatics and mass spectrometry challenge. PMID- 15900445 TI - Building a professional career. Initiative and meticulousness: mining, digging for the rare nuggets. PMID- 15900446 TI - Silver nanoparticle assemblies supported on glassy-carbon electrodes for the electro-analytical detection of hydrogen peroxide. AB - Electrochemical detection of hydrogen peroxide using an edge-plane pyrolytic graphite electrode (EPPG), a glassy carbon (GC) electrode, and a silver nanoparticle-modified GC electrode is reported. It is shown, in phosphate buffer (0.05 mol L(-1), pH 7.4), that hydrogen peroxide cannot be detected directly on either the EPPG or GC electrodes. However, reduction can be facilitated by modification of the glassy-carbon surface with nanosized silver assemblies. The optimum conditions for modification of the GC electrode with silver nanoparticles were found to be deposition for 1 min at -0.5 V vs. Ag from 5 mmol L(-1) AgNO3/0.1 mol L(-1) TBAP/MeCN, followed by stripping for 2 min at +0.5 V vs. Ag in the same solution. A wave, due to the reduction of hydrogen peroxide on the silver nanoparticles is observed at -0.68 V vs. SCE. The limit of detection for this modified nanosilver electrode was 2.0 x 10(-6) mol L(-1) for hydrogen peroxide in phosphate buffer (0.05 mol L(-1), pH 7.4) with a sensitivity which is five times higher than that observed at a silver macro-electrode. Also observed is a shoulder on the voltammetric wave corresponding to the reduction of oxygen, which is produced by silver-catalysed chemical decomposition of hydrogen peroxide to water and oxygen then oxygen reduction at the surface of the glassy-carbon electrode. PMID- 15900447 TI - The detection of nitrated tyrosine in neuropeptides: a MALDI matrix-dependent response. AB - Neuropeptides are a diverse class of signaling molecules that typically have one or more posttranslational modifications. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is an effective tool for identification and characterization of neuropeptides from samples as small as individual neurons. However, the detection of one particular posttranslational modification-nitrotyrosine-has been problematic because of the lability of the nitro group of nitrotyrosine under MALDI-MS conditions. The detection of nitrated tyrosine in peptide standards was dependent on the MALDI matrix used for the analysis. Specifically, sinapinic acid was the optimum matrix tested to observe this modification while it was not consistently detected with matrices such as 2,5-dihydroxybenzoic acid. Using the optimized procedures, several identified nitric-oxide-synthase positive neurons from Lymnaea stagnalis were tested to determine if the neuropeptides present were nitrated. In all cases, the nitrated form of the neuropeptide was not observed. The dependence on the sample preparation procedures of observing this particular chemical modification demonstrates the need for careful selection of sample-preparation methods with MALDI or the use of other ionization methods. PMID- 15900448 TI - Synthesis and analytical properties of micrometric biosensing lipobeads. AB - This paper describes the preparation for the first time of lipobead-based micrometric fluorescence biosensors and the optimization of their analytical properties. The study focused on the well-established urea biosensors as a model system. Fluorescence-sensing lipobeads were prepared by coating carboxyl functionalized silica microspheres with phospholipids. The enzyme urease and the pH indicator fluorescein-5-thiosemicarbazide were then attached covalently to the phospholipid membrane of the lipobeads. Urease converts urea to ammonia, which results in a pH increase in the analyte solution and to a urea concentration dependent increase in the fluorescence intensity of the sensing lipobeads. Previous fluorescence-sensing lipobeads were synthesized by coating polystyrene particles with a phospholipid membrane. The membrane was physically attached to the particles and the fluorophores were entrapped in the membrane. In this study, we prepared improved fluorescence-sensing lipobeads by utilizing covalent chemistry to bind the phospholipid membrane to the silica particles and the fluorophores to the membrane. This led to improvement in the stability of the newly developed urea-sensing lipobeads compared to previously developed miniaturized fluorescence biosensors. PMID- 15900449 TI - Individual cell migration analysis using fiber-optic bundles. AB - In this paper we describe a novel optical fiber-based technology for analyzing cell migration. Cells were labeled with a membrane-bound fluorescent dye and distributed onto a polished optical fiber bundle. When a cell passes over one of the individual fibers in the bundle, the membrane-bound dye causes a large intensity increase, which stays for a given "residence time" until the cell departs from the fiber. Residence time increases significantly upon exposure to an antimigratory drug, indicating a decrease in cell migration. This approach provides a simple migration assay and does not require sophisticated tracking software. By using optical fiber bundles containing smaller individual fibers with higher spatial resolution, this approach was employed to develop a migration assay based on subcellular imaging. The subcellular imaging platform allows for rapid analysis of migratory potential, reducing experimental time from several hours in a standard assay to 5 min using this technology. PMID- 15900450 TI - Amperometric nitrite sensor based on hemoglobin/colloidal gold nanoparticles immobilized on a glassy carbon electrode by a titania sol-gel film. AB - A novel amperometric nitrite sensor was developed based on the immobilization of hemoglobin/colloidal gold nanoparticles on a glassy carbon electrode by a titania sol-gel film. The sensor shows a pair of well-defined and nearly reversible cyclic voltammogram peaks for Hb Fe(III)/Fe(II) with a formal potential (E degrees ') of -0.370 V, and the peak-to-peak separation at 100 mV s(-1) was 66 mV vs. Ag/AgCl (3.0 M KCl) in a pH 6.9 phosphate buffer solution. The formal potential of the Hb Fe(III)/Fe(II) couple shifted linearly with pH with a slope of -50.0 mV/pH, indicating that electron transfer accompanies single-proton transportation. The sensor exhibited an excellent electrocatalytic response to the reduction of nitrite. The reduction overpotential was 0.45 V below that obtained at a colloidal gold nanoparticles/TiO2 sol-gel film-modified GCE. The linear range for nitrite determination for the sensor was 4.0 x 10(-6) to 3.5 x 10(-4) M, with a detection limit of 1.2 x 10(-6) M. The stability, repeatability and selectivity of the sensor were also evaluated. PMID- 15900451 TI - Molecularly imprinted polymers as tools for the screening of felodipine from dihydropyridine calcium antagonists by pressurized capillary electrochromatography. AB - A group of structurally similar dihydropyridine calcium antagonists (DHPs) and related compounds were used to simulate a combinatorial library. A molecularly imprinted polymer (MIP) comprising felodipine (FLD) was synthesized in situ inside the capillary for use in the separation of FLD from other DHPs by pressurized electrochromatography (pCEC). To evaluate the feasibility of using the MIP columns for the separation of FLD, parameters including pH, the applied voltages, and the effect of organic modifier were studied. The results indicated that the MIP columns demonstrated better recognition properties over a pH range of 4-6. The efficiency (plates/m) at pH 5.0 for the non-imprinted analytes was 117,000 for thiourea, 18,700 for nicarpidine, 17,300 for nisoldipine, and 14,600 for nifedipine; however, the efficiency for the imprinted analyte FLD was low, as evidenced by the broad peak, yielding only 5,100 plates/m. The column efficiency was also investigated under both micro-HPLC and pCEC conditions. PMID- 15900452 TI - Direct fluorometric analysis of a newly synthesised fluorescein-labelled marker for glomerular filtration rate. AB - There is an obvious and growing medical need for an accurate determination of kidney function in the diagnosis and management of renal diseases. The glomerular filtration rate (GFR) is the accepted gold standard measurement of kidney function. Several approaches to estimate the GFR are available, but most of them are inconvenient and, therefore, of limited acceptance. A new method of quantification with fluorescein-isothiocyanate (FITC) sinistrin (FS), a novel GFR marker, has been evaluated. The method is based on the fluorescence label of FS and can be performed with a standard fluorometer. To control the interference of protein with the fluorescence signal, a calibration function was developed. The accuracy of the fluorometric method established is comparable to the so-called "gold standard" of enzymatic determination of polyfructosan. Moreover, FS is easy to handle and requires low-cost instruments. Our results demonstrate the potential of the direct fluorometric analysis of the new FITC-labelled marker of being a precise, simple, rapid and cost-effective method for diagnosing disturbed kidney function and monitoring its treatment efficacy. The dramatic decrease in analytical effort will result in a significantly higher acceptability of GFR determination. PMID- 15900453 TI - Colored pI standards and gel isoelectric focusing in strongly acidic pH. AB - Colored, low molecular weight pI markers have been developed for isoelectric focusing (IEF) in acidic pH range. Their isoelectric points (pIs) were determined by direct measurement of the pH of the focused bands after completion of IEF on polyacrylamide gels. The practicable suitability of the proposed pI markers as pI standards for IEF was tested by applying gel IEF. The acidic pH gradient was created either by commercial synthetic carrier ampholytes or by mixture of simple buffers consisting of acids (non-ampholytes) and ampholytic buffers. By applying simple acids, it was possible to extend the acidic pH range beyond those achievable with commercial synthetic carrier ampholytes. By using an experimental arrangement without electrode electrolyte reservoirs with electrodes creating the fixed end of the gel, the strongly acidic pH gradient was stable even for prolonged focusing time. PMID- 15900454 TI - Uranium analysis in urine by inductively coupled plasma dynamic reaction cell mass spectrometry. AB - Urine uranium concentrations are the best biological indicator for identifying exposure to depleted uranium (DU). Internal exposure to DU causes an increased amount of urine uranium and a decreased ratio of 235U/238U in urine samples, resulting in measurements that vary between 0.00725 and 0.002 (i.e., natural and depleted uranium's 235U/238U ratios, respectively). A method based on inductively coupled plasma dynamic reaction cell mass spectrometry (ICP-DRC-MS) was utilized to identify DU in urine by measuring the quantity of total U and the 235U/238U ratio. The quantitative analysis was achieved using 233U as an internal standard. The analysis was performed both with and without the reaction gas oxygen. The reaction gas converted ionized 235U+ and 238U+ into 235UO2+ (m/z = 267) and 238UO2+ (m/z = 270). This conversion was determined to be over 90% efficient. A polyatomic interference at m/z 234.8 was successfully removed from the 235U signal under either DRC operating conditions (with or without oxygen as a reaction gas). The method was validated with 15 urine samples of known uranium compositions. The method detection limit for quantification was determined to be 0.1 pg U mL(-1) urine and an average coefficient of variation (CV) of 1-2% within the sample measurements. The method detection limit for determining 235U/238U ratio was 3.0 pg U mL(-1) urine. An additional 21 patient samples were analyzed with no information about medical history. The measured 235U/238U ratio within the urine samples correctly identified the presence or absence of internal DU exposure in all 21 patients. PMID- 15900455 TI - Charge-selective recognition at fibroin-modified electrodes for analytical application. AB - A novel fibroin-modified electrode with charge recognition is reported. The characteristics of silk fibroin membranes have been exploited for analytical applications. The membrane, with an isoelectric point of pH 4.5, was applied to graphite and carbon-fiber electrodes. The modified electrode was negatively charged in solutions of pH > 4.5, and so rejected anions and attracted cations. In solutions of pH < 4.5 the electrode was positively charged, and so rejected cations and attracted anions. The pH-responsive charge recognition of the modified electrode was investigated for some neurocompounds. A fibroin carbon fiber electrode was used for in-vivo determination of the concentration of the cationic neurotransmitter dopamine (DA). PMID- 15900456 TI - Pharmacokinetic detection of penicillin excreted in urine using a totally internally reflected resonance light scattering technique with cetyltrimethylammonium bromide. AB - A quantitative analysis method for penicillins including ampicillin (AmP), benzyl penicillin (BP), oxacillin (OA) and amoxycillin (AmO) is proposed that makes use of the totally internally reflected resonance light scattering (TIR-RLS) signal from the penicillin at the H2O/CCl4 interface in the presence of cetyltrimethylammonium bromide (CTMAB), and enables the pharmacokinetics of penicillin taken orally and excreted through urine to be monitored. Penicillin is coadsorbed with CTMAB at the H2O/CCl4 interface in neutral solution, resulting in the formation of ion associates that display greatly enhanced TIR-RLS signals (maximum at 368-372 nm). This enhanced TIR-RLS intensity was found to be proportional to the penicillin concentration over the range 0.2 x 10(-6) to 2.2 x 10(-6) mol L(-1), with limits of determination (3sigma) of 5.0 x 10(-8) to 7.0 x 10(-8) mol L(-1). Pharmacokinetics studies performed using the present method show that the excretion of orally-taken ampicillin through urine has a half-time of 1.05 h and an excremental quantum over 8 h of 49.3%, respectively. PMID- 15900457 TI - Development and validation of an analytical method for detection of estrogens in water. AB - An analytical procedure enabling routine analysis of four environmental estrogens at concentrations below 1 ng L(-1) in estuarine water samples has been developed and validated. The method includes extraction of water samples using solid-phase extraction discs and detection by gas chromatography (GC) with tandem mass spectrometry (MS-MS) in electron-impact (EI) mode. The targeted estrogens included 17alpha- and 17beta-estradiol (aE2, bE2), estrone (E1), and 17alpha ethinylestradiol (EE2), all known environmental endocrine disruptors. Method performance characteristics, for example trueness, recovery, calibration, precision, accuracy, limit of quantification (LOQ), and the stability of the compounds are presented for each of the selected estrogens. Application of the procedure to water samples from the Scheldt estuary (Belgium - The Netherlands), a polluted estuary with reported incidences of environmental endocrine disruption, revealed that E1 was detected most frequently at concentrations up to 7 ng L(-1). aE2 was detected once only and concentrations of bE2 and EE2 were below the LOQ. PMID- 15900458 TI - Simultaneous determination of 64 pesticides in river water by stir bar sorptive extraction and thermal desorption-gas chromatography-mass spectrometry. AB - A method for determining 68 pesticides in river water using stir bar sorptive extraction (SBSE)-thermal desorption (TD)-gas chromatography-mass spectrometry (GC-MS) is described. SBSE sampling was optimized for sample solution pH, salting out and methanol addition. Although salting-out enhanced the ability of the method to extract most of the pesticides with low absolute recoveries, the absolute recoveries of four pesticides were not improved by salting-out. The detection limits of the method for the pesticides ranged from 0.2 to 20 ng/l. Analyte recoveries from a river water sample spiked with standards at 10 and 100 ng/l were 58.5-132.0% (RSD: 1.8-15.8%) and 61.0-121.3% (RSD: 1.4-20.2%), respectively. PMID- 15900459 TI - Monitoring and effects of nicosulfuron in aquatic mesocosms: development of a simple analytical procedure and evidence for low toxicity to phytoplankton communities. AB - A new analytical procedure is proposed for monitoring nicosulfuron (sulfonylurea herbicide) in aquatic mesocosms derived from complex ecosystems. The approach is based on alternate use of the anionic and molecular forms of the pesticide during the procedure. It also takes into account the sensitivity of the molecule to hydrolysis. The procedure involves solid-phase extraction on a polystyrene divinylbenzene support followed by a conventional high-performance liquid chromatographic analysis with UV-diode array detection. Recovery tests on samples from natural waters demonstrated that the performances obtained were convenient for monitoring aquatic mesocosms (recoveries of 91+/-12% at 0.5 microg L(-1)). The method was used to monitor nicosulfuron in mesocosms initially spiked at 2 or 30 microg L(-1) . The evolution curves were compared to those obtained from mesocosms contaminated with atrazine at the same initial doses. The sensitivity of phytoplankton communities to nicosulfuron in aquatic mesocosms was found to be very limited. PMID- 15900460 TI - Analysis of volatile components from Ziziphora taurica subsp. taurica by steam distillation, superheated-water extraction, and direct thermal desorption with GCxGC-TOFMS. AB - Volatile components from the leaves of Ziziphora taurica subsp. taurica have been isolated by steam distillation, superheated-water extraction, and direct thermal desorption techniques. The volatile components were characterized by comprehensive two dimensional gas chromatography-time of flight mass spectrometry. The extraction yields from Z. taurica leaves were found to be 1.56% and 1.32% for steam distillation and superheated-water extraction respectively. The major compounds found in the volatile fractions of the leaves of Z. taurica were pulegone, terpinen-4-ol, cis-carveol, trans-carveol, and verbenone. The number of volatile components identified were 28, 30, and 41 for steam distillation, superheated-water extraction, and direct thermal desorption, respectively. PMID- 15900461 TI - Air handling in clean laboratory environments: the reason for anomalously high boron background levels. AB - The isotopic analysis of boron from material that carries only trace amounts of boron requires low boron blank levels. Large efforts are taken to keep blank levels low by purifying reagents. We performed exposure experiments and determined procedure blanks before and after the air-handling system was modified from silicate glass filters to polyethylene/polystyrole and active carbon filters. Our investigations demonstrate that the air-handling system may be the major source of boron blank in many laboratories, as the widely used silicate glass filters, which contain boron as a major component, in the form of borax and boric acid, release significant amounts of boron to the air. PMID- 15900462 TI - An environmentally friendly method for the extraction and determination of priority phenols in soils using microwave-assisted micellar extraction. AB - A non-ionic surfactant, polyoxyethylene 10 lauryl ether (POLE), was used for the microwave-assisted extraction (MAE) of priority phenolic compounds from soil samples. A central composite design was applied to optimize the extraction parameters, namely, time and power. Under the optimized conditions, the method was applied to different soil samples in order to analyze the influence of soil characteristics on the phenol extraction. Results demonstrated that most of these compounds can be recovered from the soils investigated in good yields (higher than 80%). The standard deviation is lower than 9% (n = 6) for most analytes. Validation of the method by analyzing a reference soil sample containing eight phenols and a comparison with Soxhlet extraction are also reported. PMID- 15900463 TI - Sensitive quantification of iodide by ion-exchange chromatography with electrochemical detection at a modified platinum electrode. AB - A rapid and very sensitive method for the accurate determination of free iodide in real samples is described. The method is based on anion-exchange chromatographic separation coupled with amperometric detection at a modified platinum electrode under constant applied potential (+0.85 V vs. Ag AgCl). An experimental setup with an in-line and very effective method of electrode modification is proposed using an amperometric thin-layer cross-flow detector and a flowing solution 300 mg/L of iodide; the working electrode is polarised to the limiting current for oxidation of iodide to iodine in acidic solutions with the consequent formation of an iodine-based film. The results indicated that the modified electrode exhibits high analytical response for iodide electrooxidation with good stability and long-life. The signal intensity of daily experimental sessions (8 h), during which standards and real samples were repeatedly injected, exhibits a moderate lowering (i.e. < 6%). Using a mixture of 25 mM HNO3 and 50 mM NaNO3 as an eluent phase in ion-exchange chromatography, the detection limit of iodide was estimated to be 0.5 microg/L (S/N = 3) with an injection volume of 50 microL. This method was applied successfully to quantify the iodide content of milk samples and in wastewaters as well as trace amounts in common vegetables and solutions containing high chloride levels. PMID- 15900464 TI - Comparison of mild extraction procedures for determination of plant-available arsenic compounds in soil. AB - In this work three mild extraction agents for determination of plant-available fractions of elements in soil were evaluated for arsenic speciation in soil samples. Pepper (Capsicum annum, L.) var. California Wonder was cultivated in pots, and aqueous solutions of arsenite, arsenate, methylarsonic acid, and dimethylarsinic acid, at a concentration of 15 mg As kg(-1) soil, were added at the beginning of the experiment. Control pots (untreated) were also included. Deionized water, 0.01 mol L(-1) CaCl2, and 0.05 mol L(-1) (NH4)2SO4 were used to extract the plant-available fraction of the arsenic compounds in soil samples collected during the vegetation period of the plants. Whereas in control samples the extractable arsenic fraction did not exceed 1% of total arsenic content, soil amendment by arsenic compounds resulted in extraction of larger amounts, which varied between 1.4 and 8.1% of total arsenic content, depending on soil treatment and on the extracting agent applied. Among arsenic compounds determined by HPLC ICPMS arsenate was predominant, followed by small amounts of arsenite, methylarsonic acid, and dimethylarsinic acid, depending on the individual soil treatment. In all the experiments in which methylarsonic acid was added to the soil methylarsonous acid was detected in the extracts, suggesting that the soil bacteria are capable of reducing methylarsonic acid before a further methylation occurs. No significant differences were observed between analytical data obtained by using different extraction procedures. PMID- 15900465 TI - Detection of banned meat and bone meal in feedstuffs by near-infrared microscopic analysis of the dense sediment fraction. AB - In this paper we present an alternative method for detection of meat and bone meal (MBM) in feedstuffs by near-infrared microscopic (NIRM) analysis of the particles in the sediment fraction (dense fraction (d >1.62) from dichloroethylene) of compound feeds. To apply this method the particles of the sediment fraction are spread on a sample holder and presented to the NIR microscope. By using the pointer of the microscope the infrared beam is focussed on each particle and the NIR spectrum (1112-2500 nm) is collected. This method can be used to detect the presence of MBM at concentrations as low as 0.05% mass fraction. When results from the NIRM method were compared with the classical microscopic method, a coefficient of determination (R2) of 0.87 was obtained. The results of this study demonstrated that this method could be proposed as a complementary tool for the detection of banned MBM in feedstuffs by reinforcement of the monitoring of feeds. PMID- 15900466 TI - Solid-phase ultraviolet sensing system for determination of methylxanthines. AB - In this study the use of a single continuous-flow solid-phase UV spectrophotometric sensing system for determination of methylxanthines was evaluated. Two methods were developed to determine caffeine (CF) and theophylline (TP) in pharmaceuticals and CF and theobromine (TB) in food and beverages. The sensor is based on transient and sequential retention of the analytes on a hydrophobic sensing solid zone (octadecyl silane C18 gel) and detection of their intrinsic UV absorbance. Temporary sequencing of the arrival of the analytes at the sensing zone is achieved by on-line separation of one of the analytes using a pre-column of the same particulate material, placed just before the flow cell. After TB or TP had been carried toward the sensing zone (by the appropriate carrier solution), produced its transitory signal, and been eluted by the carrier, an appropriate eluting solution (25% MeOH) was used to elute CF, which was strongly retained on the minicolumn, so that its transient signal could be recorded. The sensing zone was completely regenerated with this eluting solution, and so was ready for analysis of another sample. After selecting the most suitable conditions, the sensing system was calibrated in the range 1-16 and 1-12 mg L(-1) for CF and TP-TB, respectively, giving detection limits below 0.1 mg L( 1) with RSD values less than 3%. The usefulness of this approach has been evaluated by applying it to the determination of caffeine, theobromine, and theophylline in different samples of food, beverages, and pharmaceutical formulations. The results were in satisfactory agreement with those obtained by use of an HPLC reference method. PMID- 15900467 TI - Determination of pesticides and some metabolites in different kinds of milk by solid-phase microextraction and low-pressure gas chromatography-tandem mass spectrometry. AB - A new analytical method is proposed to determine more than 40 multiclass pesticides in different kinds of processed (whole, skimmed and powdered) and unprocessed (goat and human) milk samples using solid-phase microextraction (SPME). A comparative study between headspace (HS) and direct immersion (DI) was carried out. The effect of milk dilution and the use of acid to reduce the influence of the matrix in DI-SPME mode were also evaluated. DI of the SPME fiber into previously diluted and acidified milk samples achieved the best sensitivity results. Pesticides were determined using low-pressure gas chromatography-tandem mass spectrometry (LP-GC-MS/MS). Both of the selected techniques have been shown to be effective at reduce fat interference and can determine analytes present at very low concentrations (limits of quantification between 0.02 and 1.00 microg L( 1)). Performance characteristics such as linearity, recovery, precision, and lower limits, together with an estimation of the measurement uncertainty using validation data, are presented for each pesticide. All of the pesticides presented recovery rates of between 81 and 110% and precision values lower than 12% (expressed as the relative standard deviation). The overall uncertainty of the method was estimated at three different concentrations (10, 25 and 50 microg L(-1)) and was lower than 25.5% in all cases. The proposed analytical methodology was applied to the analysis of target pesticides in 35 samples: 15 commercial, 3 human and 17 goat milk samples. The metabolite p,p'-DDE was the compound most frequently found in both the breast and goat milk samples, at concentration levels < 20 microg L(-1). However, pesticide residues were not found in any of the other 15 commercial milk samples (skimmed, powdered and whole milk) analyzed. PMID- 15900468 TI - Simultaneous determination of hydride (Se) and non-hydride-forming (Ca, Mg, K, P, S and Zn) elements in various beverages (beer, coffee, and milk), with minimum sample preparation, by ICP-AES and use of a dual-mode sample-introduction system. AB - A method has been developed enabling direct analysis (i.e. after dilution only) of beer, instant coffee, milk, and milk powder by ICP-AES. Analysis of the beverages after dilution with a low concentration of HNO3 was used for accurate determination of essential minor and trace elements (Ca, Mg, K, P, S, and Zn). Selenium, introduced as the hydride, was determined simultaneously with the other non-hydride-forming elements using the commercial multi-mode sample-introduction system (MSIS). To obtain accurate results, however, some simple pre-treatment was needed. Analysis was also performed after microwave-assisted decomposition of the samples. Three different modes of sample-preparation, i.e. dilution only, partial decomposition (aqua regia treatment), and complete decomposition were compared. The results obtained by use of the three different sample-preparation methods were in very good agreement. Results from analysis of certified reference material (SRM 1459 non-fat milk powder) also verified the accuracy of the methods. The limit of detection obtained for Se using dual-mode sample introduction was 0.5 ng mL(-1), which corresponds to approximately 2 ng g(-1) in beer and approximately 4 ng g(-1) in coffee and milk when using the recommended procedure. PMID- 15900469 TI - Interfacing capillary electrophoresis and surface-enhanced resonance Raman spectroscopy for the determination of dye compounds. AB - The at-line coupling of capillary electrophoresis (CE) and surface-enhanced resonance Raman spectroscopy (SERRS) was optimized for the separation and subsequent spectroscopic identification of charged analytes (dye compounds). Raman spectra were recorded following deposition of the electropherogram onto a moving substrate. To this end a new interface was developed using a stainless steel needle as a (grounded) cathode. The outlet end of the CE capillary was inserted into this metal needle; CE buffer touching the needle tip served as the electrical connection for the CE separation. A translation table was used to move the TLC plate at a constant speed during the deposition. The distance between the tip of the fused silica column and the TLC plate was kept as small as possible in order to establish a constant bridge-flow, while avoiding direct contact. The dyes Basic Red 9 (BR9), Acid Orange 7 (AO7) and Food Yellow 3 (FY3) were used as test compounds. After CE separation in a 20 mM borate buffer at pH 10, after deposition, concentrated silver colloid was added to each analyte spot, followed by irradiation with 514.5 nm light from an argon ion laser to record the SERRS signal using a Raman microscope. Different types of silver colloids were tested: Lee-Meisel type (citrate), borate, and gold-coated silver. BR9 (positively charged) gave much more intense SERRS spectra than the two negatively charged dyes. For BR9 and AO7 the citrate-coated Lee-Meisel colloid yielded the most intense SERRS spectra. The CE-SERRS system was used to separate and detect the negatively charged dyes. Silver colloid and nitric acid (to improve adsorption) were added post-deposition. Even though their chemical structures are very similar, AO7 and FY3 could be readily distinguished based on their SERRS spectra. The limits of detection (S/N = 3) of the CE-SERRS system ranged from 6.7 x 10(-5) M (2.6 x 10(-12) mol injected) for FY3 down to 1.8 x 10(-6) M (7.0 x 10(-14) mol injected) for BR9. PMID- 15900470 TI - GC-IR based two-dimensional structural group analysis of petroleum products. AB - To characterize petroleum products, a method based on the combination of IR spectrometric structural group analysis, GC-simulated boiling analysis and multivariate regression techniques (PLS, PCR) has been developed. The best performance was achieved by a PLS regression model with six aliphatic and aromatic structural groups. Thereby, structural group distribution related to the boiling temperature could be obtained in order to quantify product-specific distribution parameters and to control material-conversion processes caused by biodegradation. PMID- 15900471 TI - Electrokinetic control of fluid in plastified laser-printed poly(ethylene terephthalate)-toner microchips. AB - The application of plastified laser-printed poly(ethylene terephthalate)(PET) toner microchips to capillary electrophoresis was investigated. Electroosmotic flow was observed in the direction of the cathode for the buffer system studied (phosphate, pH 3-10). Average electroosmotic mobilities of 1.71 x 10(-4) to 4.35 x 10(-4) cm(2) V(-1) s(-1) were observed from pH 3 to 10. This variation suggests that silica fillers in the toner and on the surface of the polymer dominate the zeta potential of the material, which is also confirmed by XPS measurements. Dopamine and catechol were used as model analytes for microchip electrophoresis in combination with electrochemical detection. Results show that these two analytes can be efficiently separated and detected electrochemically with the plastified laser-printed PET-toner microchips. PMID- 15900472 TI - Determination of levodopa methyl ester and its metabolites in rat serum by CZE with amperometric detection. AB - A reliable and reproducible method, capillary zone electrophoresis with amperometric detection (CZE-AD), has been developed for separation and quantification of levodopa methyl ester (LDME) and its biotransformation products levodopa (L-DOPA) and dopamine (DA) in rat serum. A carbon-disk electrode was used as working electrode. The optimum conditions for CZE detection were 50 mmol L(-1) phosphate solution at pH 7.0 as running buffer, 17 kV as separation voltage, 1.0 V (vs Ag/AgCl, 3.0 mol L(-1)) as detection potential, and sample injection for 8 s at 17 kV. The linear ranges were from 2.4 x 10(-2) to 2.2 microg mL(-1) for LDME, 2.9 x 10(-1) to 49.5 microg mL(-1) for L-DOPA, and 1.4 x 10(-2) to 1.5 microg mL(-1) for DA with correlation coefficients of 0.9997, 0.9994, and 0.9999, respectively. The detection limits for LDME, L-DOPA, and DA were 14.6, 98.0, and 9.7 ng mL(-1), respectively. Recoveries were 80.3% for LDME, 93.5% for L-DOPA, and 86.5% for DA. This method was applied to serum samples after intravenous injection of LDME and L-DOPA to rats. PMID- 15900473 TI - Determination of camptothecin and 10-hydroxycamptothecin in human plasma using polymer monolithic in-tube solid phase microextraction combined with high performance liquid chromatography. AB - A biocompatible in-tube solid-phase microextraction (SPME) device was used for the direct and on-line extraction of camptothecin and 10-hydroxycamptothecin in human plasma. Biocompatibility was achieved through the use of a poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column for extraction. Coupled to high performance liquid chromatography (HPLC) with UV detection, this on-line in-tube SPME method was successfully applied to the simultaneous determination of camptothecin and 10-hydroxycamptothecin in human plasma. The calculated detection limits for camptothecin and 10-hydroxycamptothecin were found to be 2.62 and 1.79 ng/mL, respectively. The method was linear over the range of 10-1000 ng/mL. Excellent method reproducibility was achieved, yielding RSDs of 2.49 and 1.59%, respectively. The detection limit (S/N = 3) of camptothecin was found to reach 0.1 ng/mL using fluorescence detection. The proposed method was shown to cope robustly with the extraction and analysis of camptothecin and 10-hydroxycamptothecin in plasma samples. PMID- 15900474 TI - Microdialysis sampling and high-performance liquid chromatography with chemiluminescence detection for in-vivo on-line determination and study of the pharmacokinetics of levodopa in blood. AB - A simple, reliable, and reproducible method for in-vivo on-line separation and determination of levodopa has been based on microdialysis then high-performance liquid chromatography with chemiluminescence detection. The perfusate is perfused at a flow rate of 5 microL min(-1). The concentration of levodopa in the dialysate is determined on line with a chemiluminescence system. The dialysate sample volume is approximately 20 microL. The response of the system is linearly related to the concentration of levodopa in the range 1 x 10(-8) to 1 x 10(-6) g mL(-1) (r2 = 0.9995) with a detection limit (3sigma) of 3 x 10(-9) g mL(-1) and sample throughput of 12 h(-1); RSD is 2.8% (n = 11). The method has been successfully used to study the pharmacokinetics of levodopa in vivo; the values of the pharmacokinetics parameters Cmax, AUC(0-t) and Tmax were 16.60, 20.92 ng mL(-1), and 90 min, respectively. PMID- 15900475 TI - Determination of phenazopyridine in human plasma via LC-MS and subsequent development of a pharmacokinetic model. AB - This paper describes a new LC-MS method for the determination of phenazopyridine and the subsequent development of a pharmacokinetic model for phenazopyridine in vivo. Phenazopyridine hydrochloride is a strong analgesic used in the treatment of urinary tract infections. Although it has been used as a clinical treatment for a very long time, pharmacokinetic data and suitable methods for its determination in plasma are currently lacking. The study described in this paper used high performance liquid chromatography-mass spectrometry, HPLC-MS, to determine the plasma concentrations of phenazopyridine in human subjects after oral administration. After liquid-liquid extraction, the phenazopyridine in the plasma was analyzed on a C18 column under SIM mode. A double-peak phenomenon was observed in most of the concentration-time profiles of the subjects. Although some drugs are known to cause this phenomenon, phenazopyridine has not been reported to do so. Several possible causes were analyzed in order to obtain an explanation. We proposed a two-site absorption compartment model to fit the concentration data in vivo, which has one more absorption site than the classical one-compartment model. The model describes the concentration profiles in different dose groups well and could provide an explanation for the double-peak phenomenon. The three dose groups exhibited similar model parameters and a linear pharmacokinetic process over the dose range used. PMID- 15900476 TI - Liquid chromatographic-electrospray mass spectrometric determination of cyclosporin A in human plasma. AB - A rapid, sensitive and selective liquid chromatography-mass spectrometry (LC-MS) assay has been developed for determination of cyclosporin A (CyA) in human plasma; cyclosporin B (CyB) was used as internal standard (IS). The method utilized a combination of a column-switching valve and a reversed-phase symmetry column. The mobile phase was a 25:75 (v/v) mixture of 10% aqueous glacial acetic acid and acetonitrile. Running time per single run was less than 10 min. Sample preparation included C8 SPE of human plasma spiked with the analyte and internal standard, evaporation of the eluate to dryness at 50 degrees C under N2 gas, and finally reconstitution in the mobile phase. Detection of cyclosporin A and the IS was performed in selected ion-monitoring mode at m/z 601.3 and 594.4 Da for CyA and IS, respectively. Quantitation was achieved by use of the regression equation of relative peak area of cyclosporin to IS against concentration of cyclosporin. The method was validated according to FDA guideline requirements. The linearity of the assay in the range 5.0-400.0 ng mL(-1) was verified as characterized by the least-squares regression line Y = (0.00268+/-1.9 x 10(-4))X+(0.00078+/-1.8 x 10(-3)), correlation coefficient, r = 0.9986+/-1.1 x 10(-3) (n = 48). Intra and inter-day quality-control measurements in the range 5.0-350.0 ng mL(-1) revealed almost 100% accuracy and < or = 9% CV for precision. The mean absolute recovery of CyA was found to be 84.01+/-9.9% and the respective relative recovery was 100.3+/-9.19. The limit of quantitation (LOQ) achieved was 5 ng mL(-1). Eventually, stability testing of the analyte and IS in plasma or stock solution revealed that both chemicals were very stable when stored for long or short periods of time at room temperature or -20 degrees C. PMID- 15900477 TI - Determination of total cysteamine in human plasma in the form of its 2-S quinolinium derivative by high performance liquid chromatography. AB - Cysteamine (mercaptamine) can be determined in plasma by liquid chromatography with ultraviolet detection after precolumn derivatization. The plasma is reduced with sodium borohydride in order to convert disulfides to thiols, and derivatized with 2-chloro-1-methylquinolinium tetrafluoroborate. The 2-S-quinolinium derivative of cysteamine is then separated from other thiols derivatives present in the plasma, and quantitated using high-performance liquid chromatography and then detection at 355 nm. Peaks from the main plasma thiols cysteine, cysteinylglycine, glutathione and homocysteine are also observed and can be measured as needed. The cystamine standards added to the plasma before the reduction step show that the response of the detector is linear within the range studied, from 0.1 to 40 micromol/L plasma. The imprecisions at the bottom and the top of the calibration range were 11.17 and 0.8% and the inaccuracies 8.64 and 1.50%, respectively, and the lower limit of quantitation was 0.1 nmol cysteamine in 1 ml of plasma. PMID- 15900478 TI - Liquid chromatographic/mass spectrometric investigation on the reaction products in the peroxidase-catalyzed oxidation of o-phenylenediamine by hydrogen peroxide. AB - Mass spectrometric evidence was obtained to confirm that the main reaction product of the horseradish peroxidase (POD)-catalyzed oxidation of o phenylenediamine (OPD) by hydrogen peroxide is 2,3-diaminophenazine. Although this reaction is one of the most widespread detection schemes in enzyme-linked immunosorbent assays (ELISAs), the literature data on the identity of the reaction product(s) have been strongly contradictory throughout the last few decades. Liquid chromatography with UV/Vis and mass spectrometric detection as well as exact mass measurements after LC fraction collection have led to the unambiguous identification of 2,3-diaminophenazine as main reaction product. 2,2' Diaminoazobenzene, which is frequently described in other publications to be the major reaction product, was not detected at all. PMID- 15900479 TI - Determination of Iron in Caco-2 cells by ET-AAS. AB - An electrothermal atomic absorption spectrometric method (ET-AAS) was developed for the direct determination of iron in intestinal Caco-2 cells after studying cell viability and proliferation using 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT test). Zeeman background correction and end-capped graphite tubes with L'vov platforms were used. Samples were dissolved in dimethylsulfoxide (DMSO) and pipetted directly into the graphite tube. The preashing, pretreatment and atomization steps were optimized. The temperatures selected were 600, 1200, and 2100 degrees C, respectively. Stability measurements were performed using iron standard solutions in DMSO on the one hand and acidified cell solutions on the other. Direct measurement and standard addition were compared in order to determine possible influences of the matrix. The low detection limit of the ET-AAS method (1.3 microg/L or 3.3 microg/g) combined with the small sample quantities required are ideal for the determination of iron in cells due to the low iron content and the limited growth area of the cells. The method was developed for iron uptake studies for toxicological purposes. PMID- 15900480 TI - Vitreous collagen metabolism before and after vitrectomy. AB - PURPOSE: To assess vitreous metabolism by measuring C-propeptide levels of type II procollagen (pCOL-II-C) and hyaluronan levels in the vitreous and in the vitreous fluid after vitrectomy for macular hole. METHODS: We obtained 1-ml vitreous samples during vitrectomy from 34 patients (34 eyes) with a macular hole (age range 50-77 years, mean 64 years). After vitrectomy, we performed fluid-air exchange in six eyes because of unresolved macular holes and collected 4-ml fluid samples. Gel-filtration high-performance liquid chromatography (HPLC) was used to determine the molecular weight of pCOL-II-C in the samples. The pCOL-II-C level was measured by sandwich enzyme immunoassay and hyaluronan by sandwich binding protein assay. RESULTS: HPLC showed that pCOL-II-C in the vitreous samples corresponded to purified pCOL-II-C from cartilage. The vitreous pCOL-II-C level (4.7+/-0.3 ng/ml) was similar to reported synovial fluid levels. In six eyes that underwent fluid-air exchange, pCOL-II-C in the fluid samples remained at a level similar to that in the vitreous samples, while hyaluronan levels in the fluid samples were significantly lower than in the vitreous samples. CONCLUSIONS: The molecular weight and concentrations of pCOL-II-C in the vitreous are similar to those in joint fluid. In patients with a macular hole, type II procollagen may be secreted persistently into the vitreous cavity before and after vitrectomy. PMID- 15900481 TI - Chiral recognition of aromatic compounds by beta-cyclodextrin based on bimodal complexation. AB - The chiral recognition of the selected aromatic chiral compounds by native beta cyclodextrin (beta-CD) based on bimodal complexation was studied using a flexible docking algorithm FDOCK. A quantitative empirical free energy relationship model was employed to predict the complex stability constants and the preferred binding modes. The results showed that the calculated complex stability constants are in good agreement with the experimental data. Furthermore, the main force responsible for host-guest complexation is the van der Waals force and the chiral molecules are completely included into the beta-CD cavity. The chiral recognition for the selected aromatic chiral compounds is the result of the van der Waals force counterbalancing with the other effects, such as the electrostatic interaction and the hydrophobic effect. PMID- 15900482 TI - Pattern computation in neural communication systems. AB - Biological data suggests that activity patterns emerging in small- and large scale neural systems may play an important role in performing the functions of the neural system, and in particular, neural computations. It is proposed in this paper that neural systems can be understood in terms of pattern computation and abstract communication systems theory. It is shown that analysing high-resolution surface EEG data, it is possible to determine abstract probabilistic rules that describe how emerging activity patterns follow earlier activity patterns. The results indicate the applicability of the proposed approach for understanding the working of complex neural systems. PMID- 15900483 TI - Computational theories on the function of theta oscillations. AB - Neural rhythms can be studied in terms of conditions for their generation, or in terms of their functional significance. The theta oscillation is a particularly prominent rhythm, reported to be present in many brain areas, and related to many important cognitive processes. The generating mechanisms of theta have extensively been studied and reviewed elsewhere; here we discuss ideas that have accumulated over the past decades on the computational roles it may subserve. Theories propose different aspects of theta oscillations as being relevant for their cognitive functions: limit cycle oscillations in neuronal firing rates, subthreshold membrane potential oscillations, periodic modulation of synaptic transmission and plasticity, and phase precession of hippocampal place cells. The relevant experimental data is briefly summarized in the light of these theories. Specific models proposing a function for theta in pattern recognition, memory, sequence learning and navigation are reviewed critically. Difficulties with testing and comparing alternative models are discussed, along with potentially important future research directions in the field. PMID- 15900484 TI - A field-theoretic approach to understanding scale-free neocortical dynamics. AB - A mesoscopic field-theoretic approach is compared with neural network and brain imaging approaches to understanding brain dynamics. Analysis of high spatiotemporal resolution rabbit electroencephalogram (EEG) reveals neural fields in the form of spatial patterns in amplitude (AM) and phase (PM) modulation of gamma and beta carrier waves that serve to classify EEGs from trials with differing conditioned stimuli (CS+/-). Paleocortex exemplified by olfactory EEG has one AM-PM pattern at a time that forms by an input-dependent phase transition. Neocortex shows multiple overlapping AM-PM patterns before and during presentation of CSs. Modeling suggests that neocortex is stabilized in a scale free state of self-organized criticality, enabling cooperative domains to form virtually instantaneously by phase transitions ranging in size from a few hypercolumns to an entire hemisphere. Self-organized local domains precede formation of global domains that supervene and contribute global modulations to local domains. This mechanism is proposed to explain Gestalt formation in perception. PMID- 15900487 TI - Functional IFNG polymorphism in intron 1 in association with an increased risk to promote sporadic breast cancer. AB - Interferon (IFN)-gamma is an important Th1 cytokine, which plays a role in immune surveillance and anti-tumor activity. A case-control study involving 54 sporadic breast cancer patients and 144 healthy controls was carried out to explore if the genotype variation of a proposed non-specific enhancer element with a dinucleotide (CA)n repeat in intron 1 has a role in the susceptibility to promote sporadic breast cancer. Genotype analysis carried out by single-strand length polymorphism and confirmed by sequencing showed an increased frequency of (CA)12 allele (P<0.001) and decreased frequencies of (CA)15 (P<0.01) and (CA)>15 (p<0.001) alleles in sporadic breast cancer patients as compared to controls. Further, in vitro reporter assays for (CA)12 and (CA)15 alleles suggested these to be associated with decreased and increased expressions, respectively, suggesting the (CA)12/(CA)12 background to act as one of the factors that could lead to low production of IFN-gamma. The study concludes that such genetic background for a proposed non-specific enhancer element with (CA)n repeat within intron 1 of the IFNG gene might put the individuals with this genotype at higher risk to promote the development of sporadic breast cancer due to a resultant compromised immune surveillance. PMID- 15900488 TI - The HLA-G genotype is associated with IL-10 levels in activated PBMCs. AB - Human leukocyte antigen (HLA)-G is an MHC class Ib molecule that is expressed at the feto-maternal interface during pregnancy. However, recent results have also shown that it may have important functions as an immuno-modulatory factor in adult life. Differences in the pattern of alternative splicing and in the stability of HLA-G mRNA transcripts have been associated with HLA-G polymorphisms, especially a 14 bp deletion/insertion polymorphism in the 3' untranslated region of the HLA-G gene. We have investigated the secretion of HLA G5/soluble HLA-G1 and interleukin-10 (IL-10) in lipopolysaccharide (LPS) activated peripheral blood mononuclear lymphocytes (PBMCs) in relation to the HLA G 14 bp genotype. No HLA-G5/sHLA-G1 could be detected in the non-activated control PBMC culture media, and there were no significant differences among the three HLA-G 14 bp genotypes regarding IL-10 concentrations. In LPS-activated PBMC cultures, no significant differences among the three HLA-G 14 bp genotypes regarding HLA-G5/sHLA-G1 concentrations were observed. However, this was in contrast to the IL-10 levels (P=0.0004, Kruskal-Wallis test). The +14/+14 bp PBMC samples expressed higher levels of IL-10 when compared to the -14/+14 bp genotype and the -14/-14 bp genotype. Interestingly, the IL-10 G/G polymorphism at position -1082 was more frequent in the +14/+14 bp genotype (P=0.024, chi2 test). These results support an autocrine loop between HLA-G5/sHLA-G1 and IL-10 expression in activated PBMCs, which may result in higher IL-10 levels in +14/+14 bp HLA-G genotypes. PMID- 15900489 TI - A new splicing acceptor site and poly(A)+ sequence signal within DQA1*0401 and DQA1*0501 mRNA 3'UTR contribute to increase the extraordinary diversity of mRNA isoforms. AB - In this paper, we have analysed the diversity of mRNA species generated by DQA1*0501 and DQA1*0401 alleles in homozygous B-lymphoblastoid cell lines. As we previously reported, six mRNA isoforms that differ in the 3'UTR have been identified in these cells. This diversity of mRNA species results both from the alternative use of two acceptor spliced sites and the differential selection of two poly(A)+ sequence signals by the processing machinery. In this report we describe a new acceptor sequence signal that allows generation of a new alternative spliced mRNA species. This acceptor sequence signal was also present in all of the seven DQA1 homozygous cell lines analysed. In addition, we have identified a previously undetected, non-conventional but functional, poly(A)+ sequence signal that lacks an identifiable AATAAA hexamer, one of the most important element of the core. This sequence signal allows the generation of two additional mRNA isoforms both in DQA1*0501 and DQA1*0401 homozygous cell lines but not in the others. We show that DQA1*0501 and DQA1*0401 primary transcripts can be processed into nine mRNA isoforms that differ in the 3'UTR. Finally, we summarized all the DQA1 mRNA species deriving from DQA1*0101, DQA1*0102, DQA1*0103, DQA1*0201, DQA1*0301, DQA1*0401 and DQA1*0501 alleles and shown in B lymphoblastoid cell lines. PMID- 15900490 TI - Identification of the MHC class I B locus in cynomolgus monkeys. AB - By determining the nucleotide sequences of more than 700 cDNA clones isolated from 16 cynomolgus monkeys, we identified 26 Mafa-B alleles. In addition, nine sequences with similarity to Mamu-I alleles were identified. Since multiple Mafa B alleles were found in each individual, it was strongly suggested that the cynomolgus MHC class I B locus might be duplicated and that the Mafa-I locus was derived from the B locus by gene duplication, as in the case of the Mamu-I locus of rhesus monkeys. PMID- 15900491 TI - Microsatellite typing of the rhesus macaque MHC region. AB - To improve the results gained by serotyping rhesus macaque major histocompatibility complex (MHC) antigens, molecular typing techniques have been established for class I and II genes. Like the rhesus macaque Mamu-DRB loci, the Mamu-A and -B are not only polymorphic but also polygenic. As a consequence, sequence-based typing of these genes is time-consuming. Therefore, eight MHC linked microsatellites, or short tandem repeats (STRs), were evaluated for their use in haplotype characterization. Polymorphism analyses in rhesus macaques of Indian and Chinese origin showed high STR allelic diversity in both populations but different patterns of allele frequency distribution between the groups. Pedigree data for class I and II loci and the eight STRs allowed us to determine extended MHC haplotypes in rhesus macaque breeding groups. STR sequencing and comparisons with the complete rhesus macaque MHC genomic map allowed the exact positioning of the markers. Strong linkage disequilibria were observed between Mamu-DR and -DQ loci and adjacent STRs. Microsatellite typing provides an efficient, robust, and quick method of genotyping and deriving MHC haplotypes for rhesus macaques regardless of their geographical origin. The incorporation of MHC linked STRs into routine genetic tests will contribute to efforts to improve the genetic characterization of the rhesus macaque for biomedical research and can provide comparative information about the evolution of the MHC region. PMID- 15900492 TI - Immunoglobulin light-chain genes in the rhesus macaque I: kappa light-chain germline sequences for subgroups IGKV1, IGKV and IGKV3. AB - The combined processes of immunoglobulin (IG) gene rearrangement and somatic hypermutation allow for the creation of an extremely diverse antibody repertoire. Knowledge of the germline sequence of the IG genes is required so that hypermutation and the affinity matured humoral response can be properly studied. Variable region genes can be arranged into subgroups; in humans, there are 11 IGLV subgroups and six IGKV subgroups. The rhesus macaque (Macaca mulatta) is a relevant non-human primate model for human immunological systems. A number of macaque IGHV, IGHD and IGHJ genes have already been reported, but only one light chain germline gene has been published so far. Here we report the isolation of new macaque IGKV genes by polymerase chain reaction (PCR) amplification from macaque genomic DNA using primers based on the human sequences. Twenty-eight IGKV1, 22 IGKV2 and 12 IGKV3 germline genes for the macaque were found, the open reading frames of which exhibit high homology to their human counterparts (>96, >99 and >96%, respectively). PMID- 15900493 TI - TRAV gene usage in pig T-cell receptor alpha cDNA. AB - Pig (Sus scrofa) TRA clones were isolated from cDNA libraries of total RNA from two different sources, the thymus of a 1-month-old LW strain pig and the peripheral blood lymphocytes of a 5-month-old Clawn strain pig. Among 103 complete TRA cDNA clones from both sources, 33 different TRAV genes were identified. By comparing their sequence identities against one another, these pig TRAV genes were grouped into 20 subgroups, including 13 subgroups, each containing only a single member. All of these pig subgroups gave corresponding human and mouse functional counterparts, suggesting their functional commonality. An exception was the Va01 gene segment, which lacked a functional human counterpart. The present report provides groundwork for studies on pig TRA expression. PMID- 15900494 TI - Reproductive and neurological Quaking(viable) phenotypes in a severe combined immune deficient mouse background. AB - The quaking(viable) (qkv) mutation, a spontaneous deletion of a multigenic region encompassing roughly 1 Mb at 5.9 cM on the proximal end of mouse chromosome 17, causes severe trembling in all homozygous animals and infertility in all homozygous males. Physiologically, quaking mice exhibit dysmyelination and postmeiotic spermatogenic arrest. Molecular defects in Qkv mice occur in the affected tissues, indicating the primary causes of these pathologies are cell autonomous. However, because both the reproductive and neurological defects are in immune-privileged sites and because some similar pathologies at both sites have been shown to be immune mediated, we tested whether the immune system participates secondarily in manifestation of Qkv phenotypes. The qkv mutation was bred into a severe combined immune-deficient mouse line (SCID; devoid of mature B and T cells) and penetrance of the neurological and the male sterile phenotypes was measured. Results showed that neither defect was ameliorated in the immune deficient background. We conclude that the Qkv pathologies do not likely involve a B- or T-cell-dependent response against these immune-privileged sites. PMID- 15900495 TI - Chicken TAP genes differ from their human orthologues in locus organisation, size, sequence features and polymorphism. AB - We have previously shown that in the chicken major histocompatibility complex, the two transporters associated with antigen processing genes (TAP1 and TAP2) are located head to head between two classical class I genes. Here we show that the region between these two TAP genes has transcription factor-binding sites in common with class I gene promoters. The TAP genes are also up-regulated by interferon-gamma in a similar way to mammalian TAP genes and in a way that suggests they are both transcribed from a bi-directional promoter. The gene structures of TAP1 and TAP2 differ from that of human TAPs in that TAP1 has a truncated exon 1 and TAP2 has fused exons, resulting in a much smaller gene size. The truncation of TAP1 results in the loss of approximately 150 amino acids, which are thought to be involved in endoplasmic reticulum retention, heterodimer formation and tapasin binding, compared to human TAP1. Most of the protein sequence features involved in binding ATP are conserved, with two exceptions: chicken TAP1 has a glycine in the switch region where other TAPs have glutamine or histidine, and both chicken TAP genes have serines in the C motif where mammalian TAP2 has an alanine. Lastly, the chicken TAP genes are highly polymorphic, with at least as many TAP alleles as there are class I alleles, as seen by investigating nine inbred lines of chicken. The close proximity of the TAP genes to the class I genes and the high level of polymorphism may allow co evolution of the genes, allowing TAP molecules to transport peptides specifically for the class I molecules of that haplotype. PMID- 15900496 TI - Host genes affect intestinal colonisation of newly hatched chickens by Campylobacter jejuni. AB - Campylobacter jejuni is the leading cause of food-borne gastro-enteritis and infection can be followed by severe clinical complications, such as the autoimmune neuropathy Guillain-Barre syndrome. Poultry meat is considered to be a common source of infection, with most flocks infected from 2 to 3 weeks of age. We have examined the effect of host genetics on the colonisation levels of C. jejuni in chickens. Chicks from different inbred lines were challenged with 10(7) to 10(8) cfu of C. jejuni 14N or C. jejuni 81-176 on the day of hatch and levels of bacterial colonisation measured over a period of 2-3 weeks. We consistently observed a 10- to 100-fold difference between four inbred lines in the number of C. jejuni organisms present in the cloaca or in the caeca, with the greatest differences detected between line N, which carried relatively high bacterial levels, and line 6(1), which carried relatively low numbers of bacteria. Amongst the four lines studied, major histocompatibility complex did not appear to be a major factor in determining the resistance. The difference in numbers of cloacal bacteria was observed as soon as 24 h after challenge and was still present at the end of the experiment. Lines N and 6(1) were chosen to analyse the mode of inheritance of the genetic differences in response to this infection. Challenge of progeny from reciprocal (6(1) female x N male) and (6(1) female x N male) F1 crosses and from (N female x 6(1) male) F1 female x N male backcrosses with C. jejuni 14N revealed that the difference in bacterial numbers was inherited in a manner consistent with the resistance (low bacterial numbers) controlled by a single autosomal dominant locus. These data suggest that it might be possible to identify the genes responsible by genetic mapping and candidate gene analysis. PMID- 15900497 TI - Artiodactyl emergence is accompanied by the birth of an extensive pool of diverse germline TRDV1 genes. AB - Molecular cloning of cDNA from gamma/delta T cells has shown that in sheep, the variable domain of the delta chain is chiefly determined by the expression of the TRDV1 subgroup, apparently composed of a large number of genes. There are three other TRDV subgroups, but these include only one gene each. To evaluate the extent and the complexity of the genomic TRDV repertoire, we screened a sheep liver genomic library from a single individual of the Altamurana breed and sheep fibroblast genomic DNA from a single individual of the Gentile di Puglia breed. We identified a total of 22 TRDV1 genes and the TRDV4 gene. A sequence comparison between germline and the rearranged genes indicates that, in sheep, the TRDV repertoire is generated by the VDJ rearrangement of at least 40 distinct TRDV1 genes. All germline TRDV1 genes present a high degree of similarity in their coding as well as in 5' and 3' flanking regions. However, a systematic analysis of the translation products reveals that these genes present a broadly different and specific repertoire in the complementarity-determining regions or recognition loops, allowing us to organize the TRDV genes into sets. We assume that selection processes operating at the level of ligand recognition have shaped the sheep TRDV germline repertoire. A phylogenetic study based on a sequence analysis of the TRDV genes from different mammalian species shows that the diversification level of these genes is higher in artiodactyl species compared to humans and mice. PMID- 15900498 TI - MHC promoter polymorphism in grey wolves and domestic dogs. AB - A functional immune system requires a tight control over major histocompatibility complex (MHC) gene transcription, as the abnormal MHC expression patterns of severe immunodeficiency and autoimmune diseases demonstrate. Although the regulation of MHC expression has been well documented in humans and mice, little is known in other species. In this study, we detail the level of polymorphism in wolf and dog MHC gene promoters. The promoter regions of the DRB, DQA and DQB locus were sequenced in 90 wolves and 90 dogs. The level of polymorphism was high in the DQB promoters, with variation found within functionally relevant regions, including binding sites for transcription factors. Clear associations between DQB promoters and exon 2 alleles were noted in wolves, indicating strong linkage disequilibrium in this region. Low levels of polymorphism were found within the DRB and DQA promoter regions. However, a variable site was identified within the T box, a TNF-alpha response element, of the DQA promoter. Furthermore, we identified a previously unrecognised 18-base-pair deletion within exon 1 of the DQB locus. PMID- 15900499 TI - Regulation of immunoglobulin gene transcription in a teleost fish: identification, expression and functional properties of E2A in the channel catfish. AB - The function of the transcriptional enhancer, Emu3', of the IgH locus of the channel catfish, Ictalurus punctatus, involves the interaction of E-protein and Oct family transcription factors. The E-proteins [class I basic helix-loop-helix (bHLH) family] are encoded in mammals by three genes: E2A (of which E12/E47 are alternatively spliced products), HEB, and E2-2. An E2A homologue has been identified in a catfish B-cell cDNA library and contains regions homologous to the bHLH and activation domains of mammalian and other vertebrate E2A proteins. E2A message is widely expressed, being readily detected in catfish B cells, T cells, kidney, spleen, brain, and muscle. Its expression is lower than that previously observed for TF12/CFEB, the catfish homologue of HEB. E2A strongly activated transcription of a muE5 motif-dependent construct in catfish B cells, and also activated transcription from the core region of the catfish IgH enhancer (Emu3') in a manner dependent on the presence of the muE5 site. Catfish E2A, expressed in vitro, bound the muE5 motif present in the core region of Emu3'. These results document the conservation of structure and function in vertebrate E2A and suggest a potential role of E2A in driving expression of the IgH locus at the phylogenetic level of a teleost fish. PMID- 15900500 TI - Structural and functional characterisation of the Toll like receptor 9 of Aotus nancymaae, a non-human primate model for malaria vaccine development. AB - In the absence of suitable rodent animal models for Plasmodium falciparum malaria, the efficacy testing of asexual blood-stage vaccine candidates in Aotus nancymaae represents a tool to select between different formulations before conducting expensive human clinical trials. CpG oligonucleotides (ODN) specifically promote the production of pro-inflammatory and Th1-type cytokines and they enhance the immunogenicity of co-administered antigens. Toll like receptor 9 (TLR-9) binds directly and sequence-specifically to single-stranded un methylated CpG-DNA mediating the biological effects of CpG ODN. We cloned and functionally characterised the TLR-9 cDNA of A. nancymaae. The cDNA encompassed 3,099 bp predicted to code for 1,032 amino acid residues. Results of homology searches to human TLR-9 suggested that the receptor is 93 and 94% identical at the nucleotide and amino acid sequence levels, respectively. Stimulation of splenocytes of A. nancymaae with CpG ODN resulted in proliferative responses in all animals analysed. FACS analysis of cultures incubated with CpG ODN 2006 indicated that the B cell marker CD20 was up-regulated consistent with B cell activation. The high level of sequence conservation of Aona-TLR-9 reinforces the suitability of A. nancymaae as animal model for malaria subunit vaccine development. PMID- 15900501 TI - Single nucleotide polymorphisms within the promoter region of the rhesus monkey tumor necrosis factor-alpha gene. AB - The human TNF-alpha gene is characterized by several single nucleotide polymorphisms (SNPs) in its promoter region, in part having been shown to influence TNF-alpha expression and susceptibility to various diseases. The rhesus macaque is widely used as an animal model for a variety of TNF-alpha associated pathological conditions, but little is known about genetic variation within the TNF-alpha promoter region. In order to check for such polymorphisms, primers based on rhesus sequence within 5' UTR were designed and used to amplify a approximately 1 kb product from genomic DNA of 29 animals. Sequencing and cloning revealed a total of 11 polymorphisms leading to five different haplotypes. PMID- 15900502 TI - Fas has a role in cerebral malaria, but not in proliferation or exclusion of the murine parasite in mice. AB - We examined the susceptibility of murine Fas-deficient mutants to malaria infection in order to investigate the role of Fas in an experimental murine model of cerebral malaria (CM). We infected mice of B6 and CBA wild-type and mutant backgrounds with Plasmodium berghei ANKA. The incidence of CM in the mutant mice (B6-lpr, CBA-lprcg) was decreased by about 50% compared with wild-type control strains at 2 weeks after infection. We did not observe significant differences of parasitemia during a murine malaria infection with nonlethal Plasmodium yoelii 17XNL between wild-type and lymphoproliferative (lpr) mutant mice of C3H and MRL genetic backgrounds, although B6-lpr mice exhibited significantly higher parasitemia than did B6 mice 12 to 18 days after infection. These results suggest Fas has a possible role in CM but may not play a major role in the proliferation or exclusion of a murine malaria parasite in a nonlethal infection. PMID- 15900503 TI - Dioxin: a review of its environmental effects and its aryl hydrocarbon receptor biology. AB - A highly persistent trace environmental contaminant and one of the most potent toxicants known is dioxin (2,3,7,8-tetrachlorodibenzo-para-dioxin or TCDD). TCDD induces a broad spectrum of biological responses, including induction of cytochrome P-450 1A1 (CYP1A1), disruption of normal hormone signaling pathways, reproductive and developmental defects, immunotoxicity, liver damage, wasting syndrome, and cancer. Its classification was upgraded from "possible human carcinogen" (group 2B) to "human carcinogen" (group 1) by the International Agency for Research on Cancer (IARC) in 1997. Exposure to TCDD may also cause changes in sex ratio, and tumor promotion in other animals. Because of the growing public and scientific concern, toxicological studies have been initiated to analyze the short- and long-term effects of dioxin. TCDD brings about a wide variety of toxic and biochemical effects via aryl hydrocarbon receptor (AhR) mediated signaling pathways. Essential steps in this adaptive mechanism include AhR binding of ligand in the cytoplasm of cells associated with two molecules of chaperone heatshock protein (Hsp90) and AhR interactive protein, translocation of the receptor to the nucleus, dimerization with the Ah receptor nuclear translocator, and binding of this heterodimeric transcription factor (present in CYP1A) to dioxin-responsive elements upstream of promoters that regulate the expression of genes involved in xenobiotic metabolism. PMID- 15900504 TI - Seasonal thermoregulatory responses in mammals. AB - This study examined the proportional seasonal winter adjustments of total and mass-specific basal power (watts and watts g-1, respectively), thermal conductance (watts g-1 degrees C-1), non-shivering thermogenesis capacity (ratio of NST/basal power), body temperature ( degrees C), and body mass (g) of mammals. The responses are best summarized for three different body size classes; small mammals (<100 g), intermediate-sized mammals (0.1-10 kg), and large mammals (>10 kg). The principal adjustments of the small mammals center on energy conservation, especially the Dehnel Effect, the winter reduction in body size of as much as 50%, accompanied by reductions in mass-specific basal power. On average, these reductions reduce the total basal power approximately in direct proportion to the mass reductions. Reductions in mass-specific basal power are matched by concomitant reductions in conductance to maintain the setpoint body temperature during winter. The overall thermoregulatory adjustments in small mammals serve to (a) lower overall winter power consumption, (b) maintain the setpoint body temperature, and (c) lower the lower critical limit of thermoneutrality and hence thermoregulatory costs. In intermediate-size mammals, the seasonal response is centered more on increasing thermogenic capacity by increasing basal power and NST capacity, accompanied by predictable and large reductions in conductance. The Dehnel effect is negligible. Very large mammals undergo the largest reductions in total and mass-specific basal power and conductance. However, there are too few data to resolve whether the reductions in total basal power can be attributed to the Dehnel effect, because the moderate decreases in body mass may also be caused by nutritional stress. Apart from the seasonal changes in basal power, these observations are consistent with the predictions of Heldmaier's seasonal acclimatization model. PMID- 15900505 TI - Adrenoceptor heterogeneity in the ruminal epithelium of sheep. AB - The pre-gastric rumen of sheep plays a crucial role in the fermentation of nutrients and in the absorption of nutrients and minerals. Adrenaline has been shown previously to increase ruminal absorption of glucose and water. The present study was intended to elucidate whether ruminal ion transport is also altered by adrenaline. In Ussing chambers, changes of I(sc) were recorded in isolated ovine ruminal epithelia after the serosal additions of adrenoceptor agonists or antagonists. I(sc) increased after the addition of adrenaline (10(-4) M) or clonidine (alpha2-agonist, 10(-4) M), but decreased after the addition of isoproterenol (beta-agonist, 10(-4) M) or terbutaline (beta2-agonist, 10(-5) M). The effect of adrenaline on I(sc) was augmented by the adrenoceptor antagonists prazosin (alpha(1), 10(-4) M) and bupranolol (beta, 10(-6) M), but inversed by yohimbine (alpha(2), 10(-5) M). Adrenaline induced an increase in Na+ net flux across the epithelium that was larger than the increase in equivalent current flow. It is concluded that adrenaline differentially regulates ion transport across the ruminal epithelium via alpha1-, alpha2-, and beta2-receptors. The main effect is a stimulation of electroneutral and electrogenic Na+ absorption. This stimulated Na+ absorption might be causative of increased water absorption from the rumen as described previously. PMID- 15900506 TI - Characterization of single L-type Ca2+ channels in myocytes isolated from the cricket lateral oviduct. AB - The single Ca2+ channel activity was obtained from cell-attached patch recordings with the use of pipettes filled with 100 mM Ba2+ as the charge carrier in myocytes isolated from the lateral oviduct of cricket Gryllus bimaculatus. The following results were obtained. (1) The channel had a unitary conductance of 18 pS. (2) The open time histogram of the channel could be fitted with a single exponential while the closed time histogram could be fitted with the sum of two exponentials, suggesting that there are at least one open state and two closed states for this channel. (3) The open probability of the channel increased with increasing membrane depolarization. (4) The mean current reconstructed by averaging individual current trace responses inactivated slowly and the current voltage relationship for the peak mean current showed a bell-shaped relation. (5) The dihydropyridine (DHP) Ca2+ antagonist, nifedipine, reduced the mean current by increasing the proportion of "blank" sweeps. On the other hand, the DHP Ca2+ agonist, Bay K 8644, increased the mean current by increasing the mean open-times of the channel. These results confirm a presence of DHP-sensitive L-type Ca2+ channel in myocytes isolated from the lateral oviduct of cricket G. bimaculatus. PMID- 15900507 TI - Examining the potential for nutritional stress in young Steller sea lions: physiological effects of prey composition. AB - The effects of high- and low-lipid prey on the body mass, body condition, and metabolic rates of young captive Steller sea lions (Eumetopias jubatus) were examined to better understand how changes in prey composition might impact the physiology and health of wild sea lions and contribute to their population decline. Results of three feeding experiments suggest that prey lipid content did not significantly affect body mass or relative body condition (lipid mass as a percent of total mass) when sea lions could consume sufficient prey to meet their energy needs. However, when energy intake was insufficient to meet daily requirements, sea lions lost more lipid mass (9.16+/-1.80 kg+/-SE) consuming low lipid prey compared with eating high-lipid prey (6.52+/-1.65 kg). Similarly, the sea lions lost 2.7+/-0.9 kg of lipid mass while consuming oil-supplemented pollock at maintenance energy levels but gained 5.2+/-2.7 kg lipid mass while consuming identical energetic levels of herring. Contrary to expectations, there was a 9.7+/-1.8% increase in metabolism during mass loss on submaintenance diets. Relative body condition decreased only 3.7+/-3.8% during periods of imposed nutritional stress, despite a 10.4+/-4.8% decrease in body mass. These findings raise questions regarding the efficacy of measures of relative body condition to detect such changes in nutritional status among wild animals. The results of these three experiments suggest that prey composition can have additional effects on sea lion energy stores beyond the direct effects of insufficient energy intake. PMID- 15900508 TI - Physiological properties of the gut lumen of terrestrial isopods (Isopoda: Oniscidea): adaptive to digesting lignocellulose? AB - Since any given trait of an organism is considered to represent either an adaptation to the environment or a phylogenetic constraint, most physiological gut characteristics should be adaptive in terms of optimizing digestion and utilization of the respective food source. Among the Crustacea, the taxon Oniscidea (Isopoda) is the only suborder that includes, and essentially consists of, species inhabiting terrestrial environments, feeding on food sources different from those of most other Crustacea (i.e., terrestrial leaf litter). Microelectrodes were used to assay physiological characteristics of the gut lumen from representatives of four families of terrestrial isopods: Trichoniscus pusillus (Trichoniscidae), Oniscus asellus (Oniscidae), Porcellio scaber (Porcellionidae), and Trachelipus rathkii (Trachelipodidae). Microsensor measurements of oxygen pressure (Clark-type oxygen microelectrodes) revealed that O2-consuming processes inside the gut lumen created steep radial oxygen gradients. Although all guts were oxic in the periphery, the radial center of the posterior hindgut was micro-oxic or even anoxic in the adults of the larger species. The entire gut lumen of all examined species was strongly oxidizing (Pt microelectrodes; apparent redox potential, Eh: +600-700 mV). Such conditions would allow for the coexistence of aerobic and anaerobic microorganisms, with both oxidative and fermentative activities contributing to digestion. Although bacterial O2 consumption was also observed in the midgut glands (hepatopancreas), they remained entirely oxic, probably owing to their large surface-to-volume ratio and high oxygen fluxes across the hepatopancreatic epithelium into the gland lumen. Measurements with pH microelectrodes (LIX-type) showed a slight pH gradient from acidic conditions in the anterior hindgut to neutral conditions in the posterior hindgut of O. asellus, P. scaber and T. rathkii. By contrast, the pH in the hindgut lumen of T. pusillus was almost constant. We discuss to what extent these physiological characteristics may be adaptive to the digestion of terrestrial food sources that are rich in lignocellulose. PMID- 15900509 TI - Variation in the fatty acid composition of blubber in Cape fur seals (Arctocephalus pusillus pusillus) and the implications for dietary interpretation. AB - Analysis of the fatty acid (FA) composition of blubber is a valuable tool in interpreting the diet of marine mammals. This technique is based on the principle that particular FA present in prey can be incorporated largely untransformed into predator adipose tissue stores, thereby providing biochemical signatures with which to identify prey species. Several studies of phocid seals and cetaceans have documented vertical stratification in the FA composition of blubber such that inferences about diet may vary greatly depending on the layer of the blubber that is analysed. It is not known whether blubber in otariid seals (fur seals and sea lions) also displays vertical stratification in FA composition. Furthermore, it is not known whether the FA composition of blubber is uniform in these species. In the present study, the vertical and regional variation in FA composition of blubber was investigated in seven adult female Cape fur seals (Arctocephalus pusillus pusillus). The proportion of monounsaturated fatty acids (MUFA) was greater in the outer (43.6+/-1.3%) than inner portion (40.9+/-1.2%; t(20)=5.59, P<0.001) whereas the proportions were greater in the inner than outer portions for saturated fatty acids (23.6+/-0.5% and 21.9+/-0.6%, respectively, t(20) = 5.31, P<0.001) and polyunsaturated fatty acids (PUFA, 35.5+/-0.7% and 34.5+/-0.7%, respectively, t(20) = 3.81, P < 0.001). There was an inverse relationship between MUFA and PUFA in the blubber, independent of sampling location. In addition, with the exception of the inner portion from non-lactating females, blubber from the mammary area had the highest proportions of 18:1omega9c and total MUFA, followed by blubber from the rump and neck, suggesting that the deposition and mobilisation of blubber lipids may not be uniform around the body in otariid seals. These results support the need for blubber tissue to be sampled from the same site on animals, and to the full depth of the blubber layer, to minimise variation in FA profiles that could occur if different sites and depths were sampled. Such standardisation of sampling will further aid in interpreting diet in otariid seals using the FA Signature Analysis approach. PMID- 15900510 TI - [3H]LY334370, a novel radioligand for the 5-HT1F receptor. I. In vitro characterization of binding properties. AB - [(3)H]LY334370 was developed as a radioligand to study the characteristics of this compound's interaction with the 5-HT(1F) receptor. Monovalent or divalent cations did not enhance the binding of [(3)H]LY334370 to the cloned human 5 HT(1F) receptor. In the presence of MgCl(2), the time to reach equilibrium was approximately 2 h, while in its absence equilibrium was reached in less than 1 h. [(3)H]LY334370 had high affinity for the cloned human 5-HT(1F) receptor (K(d)=0.446 nM) and the 5-HT(1F) receptor in rat brain (K(d)=0.388 nM). The expression density of 5-HT(1F) receptors, as determined by binding to homogenates of cortical regions from rat, was low (B(max)=79.1 fmol/mg protein). There was a statistically significant correlation between the apparent pK(i) for inhibition of [(3)H]LY334370 binding and the pEC(50) for stimulation of [(35)S]GTPgammaS binding to homogenates of cells expressing the cloned human 5-HT(1F) receptor. In addition, there was a statistically significant correlation between the apparent pK(i) for inhibition of [(3)H]LY334370 binding to the cloned human 5-HT(1F) receptor and the pID(50) for inhibition of trigeminal nerve stimulated dural plasma protein extravasation in the guinea pig. The conclusion from these studies is that [(3)H]LY334370 is a high affinity radioligand which can be used for the study of the 5-HT(1F) receptor in rat brain or in cells transformed with the human 5-HT(1F) receptor. PMID- 15900511 TI - [3H]LY334370, a novel radioligand for the 5-HT1F receptor. II. Autoradiographic localization in rat, guinea pig, monkey and human brain. AB - LY334370 is a high affinity, selective agonist at the 5-HT(1F) receptor. On this basis, the tritiated compound was examined for its utility in autoradiography to localize the 5-HT(1F) receptor in rat and guinea pig brain regions. Specific 5 HT(1F) receptor binding in rat brain was found in layers 4-5 of all cortical regions examined, as well as olfactory bulb and tubercle, nucleus accumbens, caudate putamen, parafascicular nucleus of the thalamus, medial mammillary nucleus, the CA3 region of the hippocampus, subiculum, and several amygdaloid nuclei. In guinea pig brain, the [(3)H]LY334370 binding sites were found at highest density in claustrum, but also in a layer of the cortex, caudate putamen, nucleus accumbens, thalamus, and medial mammillary nucleus. Some species differences in the distribution of the 5-HT(1F) receptor were noted. Side by side comparison of rat brain autoradiography with [(3)H]LY334370 and [(3)H]sumatriptan showed labeling in the same brain regions. Preliminary binding studies in rhesus monkey and human brain sections showed [(3)H]LY334370 binding in cortical layers 4-5, subiculum (in the monkey), and the granule cell layer of the cerebellum. These findings suggest a discrete localization of the 5-HT(1F) receptor in the rat, guinea pig, monkey and human brain, and confirms the utility of [(3)H]LY334370 as a potential tool to explore further the localization and possible functions of the 5-HT(1F) receptor. PMID- 15900512 TI - Differential inhibition by the Rho kinase inhibitor Y-27632 of the increases in contractility and Ca2+ transients induced by endothelin-1 in rabbit ventricular myocytes. AB - The role of Rho kinase activation in the regulation of cardiac contractility and Ca(2+) signaling remains unclear, whereas its role in smooth muscle regulation has been well documented. To study the potential role of Rho kinase in the regulation of cardiac contractility and Ca(2+) transients induced by endothelin-1 (ET-1) and isoproterenol, we used the Rho kinase inhibitor Y-27632 in rabbit ventricular myocardium and myocytes loaded with indo-1/AM. Y-27632 (3-30 microM) inhibited significantly the baseline contractility and Ca(2+) transients. Furthermore, Y-27632 suppressed the increase in contractility and Ca(2+) transients induced by ET-1 in a concentration-dependent manner, when it was used in a concentration at which it did not affect the effects of isoproterenol via beta-adrenoceptors. In the presence of Y-27632, ET-1 increased cell shortening in the absence of an increase in Ca(2+) transients. This is an indication that the increase in myofilament Ca(2+) sensitivity induced by ET-1 is less susceptible to the inhibitory action of Y-27632. These findings imply that the Rho kinase activation may partially contribute to the ET-1-induced regulation of contractility, primarily due to an ET-1-induced increase in Ca(2+) transients in rabbit ventricular myocardium. PMID- 15900513 TI - Characterisation of (R/S)-propafenone and its metabolites as substrates and inhibitors of P-glycoprotein. AB - Digoxin is a drug with a narrow therapeutic index, which is a substrate of the ATP-dependent efflux pump P-glycoprotein. Increased or decreased digoxin plasma concentrations occur in humans due to the inhibition or induction of this drug transporter in organs with excretory function such as small intestine, liver and kidney. It is well known that serum concentrations of digoxin increase considerably in humans if propafenone is given simultaneously. However, it has not been investigated in detail whether propafenone and its metabolites are substrates and/or inhibitors of human P-glycoprotein. The aim of this study, therefore, was to investigate the P-glycoprotein-mediated transport and inhibition properties of propafenone and its major metabolites 5 hydroxypropafenone and N-desalkylpropafenone in Caco-2 cell monolayers. Inhibition of P-glycoprotein-mediated transport by propafenone and its metabolites was determined using digoxin as a P-glycoprotein substrate. No polarised transport was observed for propafenone and N-desalkylpropafenone in Caco-2 cell monolayers. However, 5-hydroxypropafenone translocation was significantly greater from basal-to-apical compared with apical-to-basal (P(app) basal-apical vs. P(app) apical-basal, 10.21+/-2.63 x 10(-6) vs. 4.34+/-1.84 x 10( 6) cm/s; P<0.01). Moreover, propafenone, 5-hydroxypropafenone and N desalkylpropafenone inhibited P-glycoprotein-mediated digoxin transport with IC(50) values of 6.8, 19.9, and 21.3 microM, respectively. In summary, whereas propafenone and N-desalkylpropafenone are not substrates of P-glycoprotein, 5 hydroxypropafenone is translocated by human P-glycoprotein across cell monolayers. In addition, propafenone and its two major metabolites 5 hydroxypropafenone and N-desalkylpropafenone are inhibitors of human P glycoprotein and therefore contribute to the digoxin-propafenone interaction observed in humans. PMID- 15900514 TI - Antiarrhythmic and electrophysiological effects of long-chain omega-3 polyunsaturated fatty acids. AB - Recent studies indicate that a diet enriched in omega-3 polyunsaturated fatty acids may prevent sudden cardiac death. The goal of the present study was to elucidate how omega-3 polyunsaturated fatty acids such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and alpha-linolenic acid (ALA; 1-20 microM) may affect the cardiac activation and repolarization pattern. For this reason, DHA, EPA or ALA was infused in spontaneously beating isolated rabbit heart (Langendorff technique) and subjected to 256 electrodes epicardial mapping. All compounds exhibited a negative inotropic and chronotropic effect. EPA and ALA, but not DHA, prolonged QTc. The dispersion was enhanced at higher concentrations (>5 microM) by DHA and less (or not affected) by the others. The total activation time, reflecting ventricular conduction, was prolonged predominantly by DHA and to a lower extent by the other drugs. Atrioventricular conduction time was slowed only by DHA and EPA. To analyze of the pattern of activation, we determined the timepoint of activation as t(dU/dt(min)) for all 256 electrodes. The beat-to-beat similarity of these patterns was moderately reduced by all drugs. Regarding antiarrhythmic activity we found that the threshold for elicitation of a ventricular extrasystole was concentration-dependently enhanced by DHA and EPA, but not by ALA. DHA dose-dependently reduced longitudinal propagation velocity V(L) and to a lower extent transverse velocity V(T). Anisotropy was not significantly changed. EPA and ALA did not exhibit a systematic effect on V(L) or V(T). These results clearly demonstrate that DHA, EPA, and ALA exhibit direct electrophysiological effects with different profiles. PMID- 15900515 TI - Potentiation of sympathetic neuromuscular transmission mediated by muscarinic receptors in guinea pig isolated vas deferens. AB - In guinea-pig isolated vasa deferentia, purinergic neurogenic contractions and responses to applied adenosine 5'-triphosphate (ATP) were potentiated by carbachol; responses to adrenergic transmission and applied noradrenaline were not. Following blockade of P2 receptors and alpha-adrenoceptors, the residual neurogenic response was massively potentiated by carbachol, suggesting the presence of a non-purinergic, non-adrenergic component. In the presence of guanethidine, carbachol had no significant effect, indicating that sympathetic transmission was the only element involved. Use of oxotremorine and selective muscarinic receptor antagonists suggested that the potentiating effect of carbachol and oxotremorine was mediated via M3 muscarinic receptors without involvement of nicotinic receptors. PMID- 15900516 TI - The effects of the neurosteroids: pregnenolone, progesterone and dehydroepiandrosterone on muscarinic receptor-induced responses in Xenopus oocytes expressing M1 and M3 receptors. AB - The neurosteroids pregnenolone, progesterone, and dehydroepiandrosterone (DHEA) occur naturally in the nervous system. They act on neural tissues, participate in neuronal signaling, and are reported to alter neuronal excitability via nongenomic mechanisms. Muscarinic receptors have important roles in neuronal functions in the brain and autonomic nervous system. In this study, we investigated the effects of pregnenolone, progesterone, and DHEA on M(1) and M(3) muscarinic receptors using the Xenopus oocyte expression system. Pregnenolone and progesterone inhibited the acetylcholine (ACh)-mediated responses of M(1) and M(3) receptors expressed in Xenopus oocytes, whereas DHEA did not. The half maximal inhibitory concentrations (IC(50)) for pregnenolone inhibition of M(1) receptor- and M(3) receptor-mediated currents were 11.4 and 6.0 microM respectively; the IC(50) values for progesterone inhibition of M(1) receptor- and M(3) receptor-mediated currents were 2.5 and 3.0 microM respectively. The selective protein kinase C (PKC) inhibitor GF109203X had little effect on the pregnenolone or progesterone inhibition of the ACh-induced currents in Xenopus oocytes expressing M(1) or M(3) receptors. The inhibitory effects of pregnenolone and progesterone were overcome at higher concentrations of ACh. Pregnenolone and progesterone inhibited the [(3)H]quinuclidinyl benzilate (QNB) binding to M(1) and M(3) receptor expressed in Xenopus oocytes, and Scatchard plot analysis of [(3)H]QNB binding revealed that pregnenolone and progesterone altered the K(d) value and the B(max), indicating noncompetitive inhibition. In conclusion, pregnenolone and progesterone inhibited M(1) and M(3) receptor functions noncompetitively by the mechanism independent of PKC and by interfering with ACh binding to the receptors. PMID- 15900520 TI - Cross-linking of polyolefins: a study by thermoporosimetry with benzene derivatives as swelling solvents. AB - The solvents o-, m-, p-xylene, p-dichlorobenzene, 1,2,4-trichlorobenzene, and naphthalene were calibrated as condensates used in the thermoporosimetry technique. Exponential relationships were found connecting the pore radii R(p) (in nm) and the freezing-point depression of the swelling solvent deltaT (in degrees C) on the one hand and the apparent energy of crystallization W(a) (in J cm(-3)) and deltaT on the other hand: R(p) = t exp[-1/(c deltaT)]; W(a) = W0 exp(deltaT/f). Pore- or mesh-size distributions can be derived from differential scanning calorimetry results by using the following equation: dV(p)/dR(p) = k{[cY(T)deltaT2]/[W(a)R(p)]}. All the numerical parameters were determined. Polyethylene and polypropylene samples, cross-linked with high-energy electrons or gamma-rays, were submitted to thermoporosimetry study. Relative mesh-size distributions, which depend on the polymer/solvent pair, were calculated for these polyolefins with o-, m-, and p-xylene as solvent and were found to be in the same sequence as those of their degrees of swelling and the irradiation doses received. PMID- 15900517 TI - Novel human alpha1a-adrenoceptor single nucleotide polymorphisms alter receptor pharmacology and biological function. AB - We identified nine naturally-occurring human single nucleotide polymorphisms (SNPs) in the alpha(1a)-adrenoceptor (alpha(1a)AR) coding region, seven of which result in amino acid change. Utilizing rat-1 fibroblasts stably expressing wild type alpha(1a)AR or each SNP at both high and low levels, we investigated the effect of these SNPs on receptor function. Compared with wild type, two SNPs (R166K, V311I) cause a decrease in binding affinity for agonists norepinephrine, epinephrine, and phenylephrine, and also shift the dose-response curve for norepinephrine stimulation of inositol phosphate (IP) production to the right (reduced potency) without altering maximal IP activity. In addition, SNP V311I and I200S display altered antagonist binding. Interestingly, a receptor with SNP G247R (located in the third intracellular loop) displays increased maximal receptor IP activity and stimulates cell growth. The increased receptor signaling for alpha(1a)AR G247R is not mediated by altered ligand binding or a deficiency in agonist-mediated desensitization, but appears to be related to enhanced receptor-G protein coupling. In conclusion, four naturally-occurring human alpha(1a)AR SNPs induce altered receptor pharmacology and/or biological activity. This finding has potentially important implications in many areas of medicine and can be used to guide alpha(1a)AR SNP choice for future clinical studies. PMID- 15900521 TI - Determination of As(III) and As(V) in oilseeds by chronopotentiometric stripping analysis: development of a method. AB - Chronopotentiometric stripping analysis (CSA) was used for selective determination of As(III) and As(V) in different oilseeds. After the optimization of experimental parameters an appropriate procedure for sample pretreatment was developed. A detection limit of 2 microg/dm3 for As(III) was obtained with an electrolysis time of 600 s. This method was used for arsenic determination in sunflower, pumpkin, and flax seed, as well as for soy flakes and almond. PMID- 15900522 TI - Regio- and stereocontrolled total synthesis of benanomicin B. PMID- 15900523 TI - Sensing nitrite through a pseudoazurin-nitrite reductase electron transfer relay. AB - Nitrite is converted to nitric oxide by haem or copper-containing enzymes in denitrifying bacteria during the process of denitrification. In designing an efficient biosensor, this enzymic turnover must be quantitatively assessed. The enzyme nitrite reductase from Alcaligenes faecalis contains a redox-active blue copper centre and a nonblue enzyme-active copper centre. It can be covalently tethered to modified gold-electrode surfaces in configurations in which direct electron transfer is possible. A surface cysteine mutant of the enzyme can be similarly immobilised on bare electroactive gold substrates. Under such circumstances, however, electron transfer cannot be effectively coupled with substrate catalytic turnover. In using either the natural redox partner, pseudoazurin, or ruthenium hexammine as an "electron-shuttle" or "conduit" between enzyme and a peptide-modified electrode surface, the coupling of electron transfer to catalysis can be utilised in the development of an amperometric nitrite sensor. PMID- 15900524 TI - Highly chemo- and enantioselective arylative cyclization of alkyne-tethered electron-deficient olefins catalyzed by rhodium complexes with chiral dienes. PMID- 15900525 TI - A modular strategy to artificial double helices. PMID- 15900526 TI - Total synthesis of avrainvillamide (CJ-17,665) and stephacidin B. PMID- 15900527 TI - A catalytic asymmetric three-component 1,4-addition/aldol reaction: enantioselective synthesis of the spirocyclic system of vannusal A. PMID- 15900528 TI - A remarkable ring contraction en route to the chartelline alkaloids. PMID- 15900529 TI - Phosphine-stabilized copper-antimony clusters: syntheses, structures, and theoretical investigations of [Cu12(SbSiMe3)6(PiPr3)6], [Cu40Sb12(PMe3)20], and [Cu45Sb16(PEt2Me)16]. PMID- 15900530 TI - Total synthesis of cavicularin and riccardin C: addressing the synthesis of an arene that adopts a boat configuration. PMID- 15900531 TI - Synthesis of the putative structure of 7-deoxycylindrospermopsin: C7 oxygenation is not required for the inhibition of protein synthesis. PMID- 15900532 TI - Epitaxial casting of nanotubular mesoporous platinum. PMID- 15900533 TI - "Catalyst-on-a-tape"-Teflon: a new delivery and recovery method for homogeneous fluorous catalysts. PMID- 15900534 TI - Inhibitors of protein tyrosine phosphatases: next-generation drugs? AB - The protein tyrosine phosphatases (PTPs) constitute a family of closely related key regulatory enzymes that dephosphorylate phosphotyrosine residues in their protein substrates. Malfunctions in PTP activity are linked to various diseases, ranging from cancer to neurological disorders and diabetes. Consequently, PTPs have emerged as promising targets for therapeutic intervention in recent years. In this review, general aspects of PTPs and the development of small-molecule inhibitors of PTPs by both academic research groups and pharmaceutical companies are discussed. Different strategies have been successfully applied to identify potent and selective inhibitors. These studies constitute the basis for the future development of PTP inhibitors as drugs. PMID- 15900535 TI - An efficient and general iron-catalyzed arylation of benzyl alcohols and benzyl carboxylates. PMID- 15900536 TI - Does a stabilising interaction favouring the Z,Z configuration of -S-N=S=N-S- systems exist? AB - The existence of the orbital interaction presented in the literature as being the cause for the stabilisation of the Z,Z configuration of Ph-S-N=S=N-S-Ph (1) and its derivatives in the crystal phase, has been investigated. The results of theoretical calculations at the DFT/B3LYP/6-311+G* level of theory suggest that such a stabilising interaction might not exist or be extremely weak and that packing forces must be the main cause of the observed Z,Z configuration in the solid. To reach this conclusion structural and energetic parameters were combined to study the bonding in these -S-N=S=N-S- systems. For the analogous Ph-Se-N=S=N Se-Ph (2) in particular the isomeric equilibrium in solution found in the variable-temperature 77Se NMR spectrum indicates that, in the gas phase or in solution, the observed Z,Z configuration is not stabilised to a greater extent than the Z,E configuration. PMID- 15900537 TI - Axial ligand effects: utilization of chiral sulfoxide additives for the induction of asymmetry in (salen)ruthenium(II) olefin cyclopropanation catalysts. PMID- 15900538 TI - Precursors of biological cofactors from ultraviolet irradiation of circumstellar/interstellar ice analogues. AB - Biological cofactors include functionalized derivatives of cyclic tetrapyrrole structures that incorporate different metal ions. They build up structural partnerships with proteins, which play a crucial role in biochemical reactions. Porphyrin, chlorin, bacteriochlorin, and corrin are the basic structures of cofactors (heme, chlorophyll, bacteriochlorophyll, siroheme, F 430, and vitamin B12). Laboratory and theoretical work suggest that the molecular building blocks of proteins (alpha-amino acids) and nucleic acids (carbohydrates, purines, and pyrimidines) were generated under prebiotic conditions. On the other hand, experimental data on the prebiotic chemistry of cofactors are rare. We propose to search directly for the pathways of the formation of cofactors in the laboratory. Herein we report on the detection of N-heterocycles and amines in the room temperature residue obtained after photo- and thermal processing of an interstellar ice analogue under high vacuum at 12 K. Among them, hexahydro-1,3,5 triazine and its derivatives, together with monopyrrolic molecules, are precursors of porphinoid cofactors. Hexahydropyrimidine was also detected. This is the first detection of these compounds in experiments simulating circumstellar/interstellar conditions. Except for 2-aminopyrrole and 2,4 diaminofuran, which were only found in 13C-labeled experiments, all the reported species were detected in both 12C- and 13C-labeled experiments, excluding contamination. The molecules reported here might be present in circumstellar/interstellar grains and cometary dust and could be detected by the Stardust and Rosetta missions. PMID- 15900539 TI - Luminescent platinum(II) terpyridyl complexes: effect of counter ions on solvent induced aggregation and color changes. AB - A series of platinum(II) terpyridyl complexes [Pt(tpy)(C triple bond C-C triple bond CH)]X, 1-X (X=OTf-; PF6-; ClO4-; BF4-; BPh4-); [Pt(tpy)(C triple bond CC6H5)]X, 2-X (X=OTf-; PF6-; ClO4-; BF4-); [Pt(tpy)(C triple bond CC6H4OCH3 4)]OTf, 3-OTf, and [Pt(4'-CH3O-tpy)(C triple bond CC6H5)]OTf, 4-OTf (tpy=2,2':6',2''-terpyridine, OTf=trifluoromethanesulfonate) were synthesized and their photophysical properties determined. Electronic absorption and emission studies showed the formation of a new band upon increasing the diethyl ether content in an acetonitrile/diethyl ether mixture. This was ascribed to the formation of complex aggregates, the solution color of which is dependent on the nature of the anions. This indicates that counter ions play an important role in governing the degree of aggregation and the extent of interactions within these aggregates. Addition of various anions to solutions of 1-OTf and 1-PF6 produced anion-induced color changes upon solvent-induced aggregation, indicating that these complexes may serve as potential colorimetric anion probes. PMID- 15900540 TI - Effect of an arbuscular mycorrhizal fungus, Glomus mosseae, and a rock-phosphate solubilizing fungus, Penicillium thomii, on Mentha piperita growth in a soilless medium. AB - Rock phosphate effect on English mint (Mentha piperita L.) grown on steamed perlite:vermiculite (1:1, v:v) substrate, with and without rock phosphate, was evaluated in greenhouse experiments. Five treatments were carried out by inoculation with an arbuscular mycorrhizal fungus Glomus mosseae and a phosphorus solubilizing microorganism Penicillium thomii. Plant aerial biomass, phosphorus concentration in plant tissue, and P available in the substrate, were evaluated upon two harvests. After the first harvest, plant aerial biomass did not show significant differences between treatments using rock phosphate as fertilizer, although P content in plants inoculated with P. thomii was higher. The second harvest revealed a higher biomass and plant tissue P content in treatments inoculated with G. mosseae. P. thomii increased P available in the substrate, whereas in the absence of G. mosseae, it did not enhance plant tissue P content. Mycorrhizal colonization was not affected by P. thomii. Microbial inoculation effect on English mint growth was also evaluated. The microbial effect was positive in all treatments when compared with the control without rock phosphate. PMID- 15900541 TI - Hyperbranching induced by cold-shock or snow-flake mutation in Neurospora crassa is prevented by addition of exogenous calcium. AB - Hyphal tip growth is a highly polarized process of cell extension, which may be affected by chemical and physical stress. Neurospora crassa exposed to cold-shock lost its polarized growth and dichotomous branches were detected. These effects were not observed in the presence of 500 mM Ca2+. We compared here the morphological pattern of a snow-flake mutant (sn) and the wild-type (wt) exposed to 4 degrees C. Hyphal morphology, nuclei, actin and microtubule distribution were analyzed. No effects on sn hyphal morphology were detected at 4 degrees C. Exogenous Ca2+ converted sn to an essentially wt appearance. The results presented here suggest that sn mutation and cold-shock treatment have affected Ca2+ influx since addition of this cation to sn (30 degrees C) and to wt (4 degrees C) maintained polarized growth and normal nuclear and microtubules distribution. PMID- 15900542 TI - Occurrence of heavy metal-resistance in microflora from serpentine soil of Andaman. AB - Serpentine soils collected from Saddle Hills, Chidyatapu and Rutland of Andaman Islands, India were analyzed for physico-chemical and microbiological characteristics and compared with those from adjacent non-serpentine localities. The serpentine soils contained high levels of nickel (1740.0-8033.4 mg/kg dry soil), cobalt (93.2-533.4 mg/kg dry soil) and chromium (302.9-4437.0 mg/kg dry soil), in addition to 62-152 g of iron and 37-60 g of magnesium per kg dry soil. Characteristically the serpentine soils showed low microbial density (6.2-11.3 x 10(6) colony forming unit/g soil) and activity (1.7-3.5 microg fluorescein/g dry soil/h) than non-serpentine outcrops. Serpentine microbial population was dominated by bacteria which represented 5.12 to 9.5 x 10(6) cfu/g of soil, while the fungal population ranged from 0.17 to 3.21 x 10(6) cfu/g of soil. A total of 342 (200 from serpentine and 142 from non-serpentine soils) isolates were compared for Ni, Co and Cr resistance. Serpentine microflora was in general, highly resistant than non-serpentine ones and showed a metal-resistance profile of Cr > Ni > Co. Amongst the serpentine isolates, 8 and 11 bacteria tolerated > 12.0 mM Ni and > 16.0 mM Cr respectively, while 6 fungal isolates showed a minimum inhibitory concentration (MIC) value > 8.0 mM Co. These 25 serpentine strains also showed co-resistance to Cu, Zn and Mn but were sensitive to Hg and Cd. The selected bacterial isolates were resistant to ampicillin, penicillin G and polymyxin B, whereas fungal strains showed resistance to amphotericin B, nystatin and fusidic acid. PMID- 15900543 TI - Developmental changes in Trichoderma viride enzymes abundant in conidia and the light-induced conidiation signalling pathway. AB - The expression of glutamic acid decarboxylase gene and the laccase activity were measured during the development of surface-cultivated Trichoderma viride mycelia in order to examine their up-regulation by light. The results show that the changes in activity of GAD induced by light observed previously are caused by transcriptional regulation of gad gene expression in both submerged mycelia and aerial mycelia after photoinduction. The expression of tga gene encoding a T. viride G(alpha) protein was found not to be up-regulated by light and was also present in the non-conidiating mutant of T. viride suggesting that this protein is not involved in the regulation of conidiation in this fungus, or that it plays a role is in later stages of conidia development. The activity of laccase was also not light-inducible and may be related to the maturation of conidia. PMID- 15900544 TI - Fungal endophytes in five medicinal plant species from Kudremukh Range, Western Ghats of India. AB - Eighteen species of endophytic fungi were isolated from bark, stem and leaf segments of five medicinal plant species growing within the Kudremukh range in the Western Ghats of India. The dominant endophytic fungal species isolated from these plant species were Curvularia clavata, C. lunata, C. pallescens and Fusarium oxysporum. The highest species richness as well as frequency of colonization of endophytic fungi was found in the leaf segments, rather than the stem and bark segments, of the host plant species. The greatest number of endophytic fungal species were found within Callicarpa tomentosa (11 species), whereas Lobelia nicotinifolia harbored the lowest number of fungal endophytes (5 species). This study provides evidence that fungal endophytes are host and tissue specific. PMID- 15900545 TI - Production of 2-methylisoborneol by Streptomyces violaceusniger and its transformation by selected species of Pseudomonas. AB - The effect of glucose, glycerol and yeast-extract (YE) on 2-methylisoborneol (MIB) production by Streptomyces violaceusniger (C4-S strain) was determined. Glycerol as a sole carbon source promoted MIB production was maximum concentration (40.65 microg/l) after 10 days incubation, while glucose favored biomass production. Increasing YE concentration from 0.1 to 1% (W/V) repressed MIB production with 62 and 83% reduction after 10 and 14 days, respectively. Transformation of this odorous compound was investigated using seven different Pseudomonas species. Action of the different Pseudomonas spp. on the C4-S extract was followed at 1 hour, 2 hours, 6 hours and 12 hours. Pseudomonas spp. 1, 2, 3 and 5 indicated a positive reaction; however, Pseudomonas spp. 4, 6 and 7 showed no effect. These data suggest that 2-methylisoborneol can be transformed by Pseudomonas spp. and the test adopted by this study can be applied for screening organisms for their ability to transform MIB. PMID- 15900546 TI - Tannic acid resistance in ruminal streptococcal isolates. AB - Tannin degrading isolates of Streptococcus spp. from rumen of non-adaptive cattle, when grown in BHI broth, were able to tolerate tannic acid upto a level of 50 g/l. An increase in lag period from 1.5 to 6 h was observed for the isolates in presence of increased concentration of tannic acid. In addition, the morphology of gram positive diplococci converted to an elongated chain of 40-50 cells with increasing tannic acid from 1 to 4%. Qualitatively, the tannase activity was found to be present in the isolates tested, indicating their potential of being a tannin degrader. PMID- 15900547 TI - Rhodium-catalyzed dehydrocoupling of fluorinated phosphine-borane adducts: synthesis, characterization, and properties of cyclic and polymeric phosphinoboranes with electron-withdrawing substituents at phosphorus. AB - The dehydrocoupling of the fluorinated secondary phosphine-borane adduct R2PH.BH3 (R = p-CF3C6H4) at 60 degrees C is catalyzed by the rhodium complex [{Rh(mu Cl)(1,5-cod)}2] to give the four-membered chain R2PH-BH2-R2P-BH3. A mixture of the cyclic trimer [R2P-BH2]3 and tetramer [R2P-BH2]4 was obtained from the same reaction at a more elevated temperature of 100 degrees C. The analogous rhodium catalyzed dehydrocoupling of the primary phosphine-borane adduct RPH2.BH3 at 60 degrees C gave the high molecular weight polyphosphinoborane polymer [RPH-BH2]n (Mw = 56,170, PDI = 1.67). The molecular weight was investigated by gel permeation chromatography and the compound characterized by multinuclear NMR spectroscopy. Interestingly, the electron-withdrawing fluorinated aryl substituents have an important influence on the reactivity as the dehydrocoupling process occurred efficiently at the mildest temperatures observed for phosphine borane adducts to date. Thin films of polymeric [RPH-BH2]n (R = p-CF3C6H4) have also been shown to function as effective negative-tone resists towards electron beam (e-beam) lithography (EBL). The resultant patterned bars were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM) and time-of flight secondary ion mass spectrometry (TOF-SIMS). PMID- 15900548 TI - Polystyrene-supported organotin dichloride as a recyclable catalyst in lactone ring-opening polymerization: assessment and catalysis monitoring by high resolution magic-angle-spinning NMR spectroscopy. AB - Dialkyltin dichloride grafted to a cross-linked polystyrene, with the formula [P H]((1-t))[P-(CH2)nSnBuCl2]t (P=[CH2CH(pC6H4)], t=the degree of functionalization, and n=6 or 11), is investigated as a recyclable catalyst in the ring-opening polymerization (ROP) of epsilon-caprolactone (CL). It is demonstrated that high resolution magic-angle-spinning (HR-MAS) NMR spectroscopy is an invaluable tool to characterize completely the supported catalyst. The 2D 1H-13C HSQC HR-MAS spectrum, in particular, allowed extensive assignment of the 1H and 13C resonances, as well as accurate measurement of the (n)J((1)H-(117/119)Sn) and (n)J((13)C-(117/119)Sn) coupling constants. 1H and 119Sn HR-MAS NMR spectroscopy is presented as a monitoring tool for catalytic processes based on organotin compounds, particularly for the investigation of the extent to which polymerization residues are observable in situ in the material pores and for the assessment of the chemical integrity and recycling conditions of the grafted catalyst. From polymerization experiments with CL, initiated by n-propanol and with [P-H]((1-t))[P-(CH2)nSnBuCl2]t of various compositions as the supported catalyst, it appears that a partial 'burst' of the polystyrene support occurs when the length of the alkyl spacer is limited to n=6, as a result of polymer chains growing within the pores of the support. However, extension of the length of the aliphatic polymethylene spacer from 6 to 11 carbon atoms preserves the support integrity and allows the production of catalyst-deprived poly(epsilon caprolactone) (PCL) oligomers. A preliminary attempt to recycle the heterogeneous catalyst has shown that very good reproducibility can be obtained, in terms of both catalyst activity and molecular-weight parameters of the as-recovered PCL polyester chains. PMID- 15900549 TI - Emergency stent graft deployment for acute aortic rupture following primary stenting for aortic coarctation. AB - We present a case of endovascular stenting for aortic recoarctation complicated by acute aortic rupture, which was successfully treated by emergency stent graft deployment. This case illustrates the potential serious complication that can occur during aortic stent dilatation and how the prompt deployment of a stent graft helped to change the clinical outcome of this life-threatening situation. PMID- 15900550 TI - The role of potassium as an ionic signal in the regulation of cyanobacterium Nostoc muscorum response to salinity and osmotic stress. AB - Spontaneously occurring thallium-resistant (Tl+ -R) mutant of diazotrophic, heterocystous cyanobacterium Nostoc muscorum is described in which Tl+ -R phenotype is the cause of defective salinity-inducible K+ uptake activity and proline uptake activity. The results indicate that mutant strain is more sensitive to salinity and osmotic stresses than its parent. The mutational inactivation of salinity-inducible proline uptake was found associated with increased sensitivity of the cyanobacterial photosynthetic O2 evolution and nitrogenase activity under salinity and osmotic stresses. It is suggested that in the cyanobacterium Nostoc muscorum K+ functions as a primary osmolyte which is associated with salinity-inducible proline uptake leading to the acquisition of salt tolerance in the cyanobacterium. PMID- 15900551 TI - Low-molecular-weight heparin vs. unfractionated heparin in percutaneous coronary intervention: a combined analysis. AB - This meta-analysis assessed the rates of the efficacy and safety endpoints with intravenous low-molecular-weight heparin (LMWH) compared with unfractionated heparin (UFH) in patients undergoing percutaneous coronary intervention (PCI). Subcutaneous LMWH has compared favorably with UFH, but limited experience exists with intravenous LMWH for immediate anticoagulation in PCI. The meta-analysis included data from eight randomized trials in which patients received LMWH (n = 1,037) or UFH (n = 978) during PCI. Seven additional nonrandomized studies/registries were analyzed to assess the efficacy and safety of LMWH during PCI. Efficacy endpoints were ischemic events (usually a composite of death, myocardial infarction, and urgent revascularization) and the safety endpoint was bleeding (major, minor, or all bleeding). In the randomized studies, LMWH was comparable with UFH in terms of efficacy (6.2% vs. 7.5%) and major bleeding (0.9% vs. 1.8%). The analysis of pooled data, randomized or not, suggests potential improved efficacy (5.8% vs. 7.6%) and reduced major bleeding (0.6% vs. 1.8%) with LMWH (n = 3,787) compared with UFH (n = 978). During PCI, intravenous LMWH without coagulation monitoring has the potential to be at least as safe and efficacious as intravenous UFH. Further studies of LMWHs in PCI are therefore required. PMID- 15900552 TI - Fluoroscopic localization of the femoral head as a landmark for common femoral artery cannulation. AB - We sought to determine the reliability of frequently used landmarks for femoral arterial access in patients undergoing cardiac catheterization. The common femoral artery (CFA) is the most frequently used arterial access in cardiac catheterization. Arterial sheath placement into the CFA has been shown to decrease vascular complications. Some authors recommend locating the inferior border of the femoral head using fluoroscopy due to the relationship of the femoral head and the bifurcation of the CFA. We performed a descriptive study in a prospective design of 158 patients undergoing catheterization from the femoral approach. A femoral angiogram was performed, and the CFA bifurcation location was recorded in relation to the inguinal ligament, middle and inferior border of the femoral head, and the inguinal skin crease. The CFA bifurcation was distal to the inguinal ligament, middle femoral head, and inferior femoral head in most patients with mean distances (cm +/- SD) of 7.5 +/- 1.7, 2.9 +/- 1.5, and 0.8 +/- 1.2, respectively. The inguinal skin crease was below the bifurcation in 78% of patients (-1.8 +/- 1.6 cm). The CFA overlies the femoral head in 92% of cases. The femoral head has a consistent relationship to the CFA, and localization using fluoroscopy is a useful landmark. PMID- 15900553 TI - Novel method for extraction of a massive refractory coronary thrombus after stent implantation. AB - Percutaneous treatment of thrombotic coronary lesions is associated with an increased incidence of complications. In particular, management of massive coronary thrombus refractory to traditional methods of removal may be especially problematic. We report a novel application of the FilterWire distal protection device to retrieve a massive refractory native coronary thrombus present after drug-eluting stent implantation. PMID- 15900554 TI - Underexpansion of sirolimus-eluting stents: incidence and relationship to delivery pressure. AB - We aimed to assess the incidence of underexpansion and the relationship between delivery pressure and expansion with sirolimus-eluting stents. Adequate stent expansion contributes to early and late improved outcomes. In 51 patients (53 lesions) with native coronary artery narrowing, balloon-expandable sirolimus eluting stents (Cypher) were serially expanded with gradual balloon inflations [14 atm, 20 atm, and in case of minimal stent cross-sectional area (CSA)/reference lumen CSA < 50% at 20 atm, postdilatation with 0.5 mm larger balloon]. Intravascular ultrasound (IVUS) imaging was performed before intervention and after each gradual balloon inflation. Stent expansion (minimal stent CSA/reference lumen CSA) was measured. Stent expansion was 72% +/- 16% after 14 atm balloon inflation, 90% +/- 18% after 20 atm balloon inflation (P < 0.001 vs. 14 atm), and 90% +/- 18% at the end of the procedure (including optional postdilatations with 0.5 mm larger balloon; P = NS vs. 20 atm). Stent expansion addressed by MUSIC criteria (all struts apposed, no tissue protrusion, and final lumen CSA > 80% of the reference or > 90% if minimal lumen CSA was < 9 mm2) was adequate in 15% of the cases after 14 atm balloon inflation, in 60% after 20 atm balloon inflation (P < 0.001 vs. 14 atm), and in 60% at the end of the procedure (P = NS vs. 20 atm). Sirolimus-eluting stent underexpansion is common when deployed at conventional pressures. Increasing balloon delivery pressure or assessing stent expansion with IVUS seems warranted in order to ensure adequate sirolimus-eluting stent deployment. PMID- 15900555 TI - On the hostile massive thrombus and the means to eradicate it. PMID- 15900556 TI - My buddy, my friend: focused force angioplasty using the buddy wire technique in an inadequately expanded stent. AB - Progressive minituarization of angioplasty hardware has resulted in a reduction of access site complications. This has limited to some degree the equipment used with small delivery systems. The buddy wire technique is a potential solution for resistant lesions with an underexpanded stent that can be used without having to upsize the introducer and guiding catheter. PMID- 15900558 TI - Small was good, will big be better? PMID- 15900559 TI - Stent-based delivery of ABT-578 via a phosphorylcholine surface coating reduces neointimal formation in the porcine coronary model. AB - Stent-based delivery of the antiproliferative and immunosuppressive macrocyclic lactone sirolimus reduces neointimal formation and restenosis by cytostatic inhibition of vascular smooth muscle cell proliferation. The objective of this study was to determine the feasibility and efficacy of stent-based delivery of ABT-578, a structurally unique macrocyclic lactone. Stainless steel balloon expandable stents were coated with thin layer of phosphorylcholine (PC) or PC with ABT-578 (10 microg/mm). Fifteen juvenile domestic pigs underwent placement of oversized bare metal (n = 15), PC (n = 8), and PC with ABT-578 (n = 9) stents in the coronary arteries. At 28 days, histology demonstrated similar mean injury scores for the control, PC-, and ABT-578-coated stents. The mean neointimal area (mm2) was significantly reduced for ABT-578 (1.70 +/- 0.47) as compared with PC (2.82 +/- 1.24) and control (2.89 +/- 1.91) stents (P < or = 0.05). The 40% reduction in neointimal area resulted in significantly less mean percent diameter stenosis for ABT-578 (19.4% +/- 4.0%) as compared with PC (30.3 +/- 12.1 %) and control (29.4% +/- 15.5%) stents (P < or = 0.03). Twelve of the 45 bare metal stent cross-sections (26.7%) exhibited a giant cell reaction, while none of the sections from the ABT-578-eluting stents had a giant cell reaction (P = 0.004). Stent-based delivery of ABT-578 via a PC surface coating inhibits neointimal formation at 28 days in the porcine coronary model. Further study is necessary to determine the dose-response and long-term effects ABT-578-eluting stents in the porcine coronary model. PMID- 15900560 TI - Determination of the concentration scaling law of the scattering coefficient of water solutions of Intralipid at 832 nm by comparison between collimated detection measurements and Monte Carlo simulations. AB - BACKGROUND AND OBJECTIVES: Intralipid (IP) is a scatterer extensively used in the building of phantoms for Biomedical Optics measurements. Recently, deviations from the linearity have been shown for the concentration scaling law of the scattering coefficient of IP water solutions at visible wavelengths. In this work this scaling law was determined at 832 nm. STUDY DESIGN/MATERIALS AND METHODS: Space resolved transmittance measurements of a laser beam at 832 nm through water solutions of IP and ink were performed and compared with the corresponding results of Monte Carlo simulations. RESULTS: The comparison provides a quadratic dependence of mu'(s) on the volume-to-volume scatterer concentration, C(IP), in the range of C(IP) values (0.0024or=40 years. AB - BACKGROUND: The 2001 Bethesda System recommends reporting benign endometrial cells (BECs) in women aged >or=40 years and considers these women at risk for endometrial pathology. The current study examines the relative risk of hyperplasia or malignancy conferred by the presence versus the absence of BECs in Papanicolaou (Pap) tests of women aged >or=40 years. METHODS: Women aged >or=40 years represented 29,177 (46.2%) of 63,202 Pap tests obtained over 1 year from a largely suburban screened population. Of these, 866 Pap tests (3%) showed BECs. Over the same 1 year period, 789 women aged >or=40 years had endometrial histologic evaluations between 14 days and 6 months following a Pap test. The Pap tests of 159 women had BECs, and 33 had atypical (n=32) or malignant (n=1) endometrial cells. The 597 remaining women, who had Pap tests without endometrial cells but who had endometrial sampling for other reasons, served as controls. RESULTS: There were nine endometrial hyperplasias (5.7%) and no adenocarcinomas in the BECs group, whereas 34 hyperplasias (5.7%) and 6 adenocarcinomas (1%) were diagnosed in the controls. These differences were not statistically significant, even after restricting the analysis to women aged >or=50 years or to women known to be postmenopausal. CONCLUSIONS: The current study found that women aged >or=40 years with BECs in their Pap tests did not have more endometrial hyperplasias or malignancies when compared with women who had endometrial sampling for reasons other than the presence of endometrial cells in a Pap test. PMID- 15900573 TI - Interleukin-4 gene, but not the interleukin-1 beta gene polymorphism, is associated with oral cancer. AB - We aimed to evaluate whether polymorphisms of the interleukin-4 (IL-4) gene promoter and intron 3 regions, and polymorphisms of the IL-1 beta gene promoter and exon 5 regions are associated with oral cancer. This study included 130 patients with oral cancer and 105 age-matched healthy controls who lived in the same area as the patients. Each genetic polymorphism was typed by polymerase chain reaction (PCR)-based restriction analysis. We then compared the genotype distribution and allelic frequencies of each polymorphism between the oral cancer patients and the controls. The CC homozygote genotype of the IL-4 gene promoter 590 region differed significantly between the patients with oral cancer and the controls (odds ratio (OR)=6.0, 95% confidence interval (CI)=1.2-30.7, chi-square test, P=0.044). No significant difference in either the genotype distribution or the allelic frequencies of the IL-1 beta gene polymorphisms was observed between patients with oral cancer and controls. The IL-4 gene -590 C/T polymorphism is associated with oral cancer and is a suitable genetic marker for screening for oral cancer. However, whether the -590 C/T polymorphism of the IL4 gene plays a role in oral cancer remains unclear. Further substantiation based on larger patient samples is needed. PMID- 15900574 TI - Effects of weekly paclitaxel or paclitaxel plus carboplatin on functionality and symptoms of geriatric patients with cancer as measured by a brief geriatric oncology module: a pilot experience. AB - BACKGROUND: To measure functionality, symptoms, and quality of life in elderly patients, a geriatric oncology module (GOM) was developed that could be rapidly and effectively administered in community oncology practices. METHODS: The GOM was validated by semi structured patient interviews. The authors tested 26 patients receiving chemotherapy with paclitaxel or paclitaxel plus carboplatin to determine the effect of drug treatment on functionality, symptoms, and quality of life (QOL). RESULTS: Chemotherapy stabilized or improved functionality, symptoms, and QOL in 75%, 86%, and 76% of patients, respectively. This GOM was easily implemented in a busy community oncology practice. CONCLUSIONS: Weekly paclitaxel and paclitaxel plus carboplatin produced measurable palliation in geriatric oncology patients. This GOM may be useful in the assessment of specific chemotherapeutic interventions in elderly patients with cancer in community oncology practices. PMID- 15900575 TI - Reflex human papilloma virus infection testing detects the same proportion of cervical intraepithelial neoplasia grade 2-3 in young versus elderly women. AB - BACKGROUND: The American Society for Colposcopy and Cervical Pathology recently recommended triage of women with atypical squamous cells of undetermined significance (ASCUS) through reflex human papilloma virus (HPV) infection testing. Because age is an important determinant of the incidence of both ASCUS and HPV infection, the current study was conducted to determine the impact of age on reflex HPV infection test performance. METHODS: The authors reviewed all liquid-based Papanicolaou tests submitted for reflex HPV infection testing to the University of Alabama Medical Center during the period from January 2002 to January 2004. To assess the impact of age on reflex HPV infection testing, women were divided into 6 age groups (<25, 25-34, 35-44, 45-54, 55-64, >64 years). Trend of different diagnostic categories with age was assessed using the chi square test for linear trend. As a measure of test performance, the proportions of cervical intraepithelial neoplasia (CIN) lesions confirmed by biopsy through reflex testing (of the total CIN lesions detected in the study population) were then compared among different age groups. RESULTS: During the study period, 25,164 ThinPrep tests were performed during the study period, with 2672 (11%) cases of ASCUS and 1371 (5%) cases of squamous intraepithelial lesions (SILs). A total of 2309 (86%) ASCUS cases were tested for HPV infection, of which 789 (34%) were positive. Biopsy specimens were obtained from 831 patients with SILs and from 316 patients with HPV+ASCUS. Of 246 patients with CIN Grade 2-3 (CIN2-3) detected on biopsy, 36 (15%) were triaged through reflex HPV infection testing and the rest were triaged through cytologic diagnosis of SILs. Despite the trend in the prevalence of HPV+ASCUS and SILs with increasing age (P < 0.05), there was no significant difference in the proportion (range, 9-23%) of CIN2-3 detected through reflex HPV infection testing in different age groups. CONCLUSIONS: Reflex HPV infection testing performed similarly for women in different age groups, including perimenopausal and postmenopausal women. PMID- 15900576 TI - Gastrointestinal stromal tumors in Iceland, 1990-2003: the icelandic GIST study, a population-based incidence and pathologic risk stratification study. AB - Gastrointestinal stromal tumor (GIST) is a newly defined clinical and pathologic entity. This study examines the whole population-based incidence of GIST as well as pathologic risk stratification schemes. All patients diagnosed in Iceland with a gastrointestinal mesenchymal tumor over the years 1990-2003 were evaluated with an immunohistochemical panel including staining for c-kit. The age-adjusted incidence of GIST was calculated. Size, mitotic rate per 50 HPF and various other pathologic parameters were evaluated. Each tumor was categorized into 1 of 4 recently defined NIH risk stratification categories. Fifty-seven of the mesenchymal gastrointestinal tumors were positive for c-kit and therefore categorized as GIST. The annual incidence for the study period is 1.1 per 100,000. The median age of patients was 65.8 years and median tumor size was 4.6 cm. Only 2 of 35 gastric tumors fall into the NIH high-risk category while half of the nongastric tumors (11 of 22) fall into this high-risk category. Eight of the 57 tumors (14%) metastasized, 7 of which were nongastric. The positive predictive value for malignant behavior of the high-risk category is 46%. The negative predictive value of low- and very-low-risk NIH category is 100%. Pathologic predictors of malignant behavior are tumor size, mitotic rate, mucosal disruption, necrosis and high cellularity. Nongastric GISTs are clearly at much higher risk of a malignant behavior than gastric GISTs. This population-based GIST study estimates the incidence of GISTs at 1.1 per 100,000 and furthermore supports the NIH consensus categories for the prediction of malignant behavior of GISTs. PMID- 15900577 TI - Clinical and immunohistologic typing of salivary duct carcinoma: a report of 50 cases. AB - BACKGROUND: Due to the low incidence of salivary duct carcinoma (SDC), only limited data in regard to the biologic behavior of this tumor and its immunohistochemical characteristics are available. The authors analyzed the clinical, molecular, and genetic profile of SDC and identified prognostic factors. METHODS: The follow-up of 50 patients with SDC was obtained and paraffin embedded tumor samples were examined immunohistochemically. In all samples, the expression of Ki-67, HER-2, and the oncoproteins p16 and p53 was examined immunohistochemically, followed by a mutation analysis of p16 and p53. The survival rate was calculated by the Kaplan-Meier method and prognostic variables were analyzed with the log-rank test. RESULTS: SDC predominantly effected male patients (66%) in their 7th decade of life. SDC mainly occurred in the parotid gland (78%; submandibular gland, 12%; minor salivary glands, 10%). Approximately two-thirds of the patients (33 of 50) presented with a T3/T4 tumor. In 28 patients (56%), cervical lymph node metastasis was present at the time of diagnosis. Local disease recurrence was observed in 48% of patients an average of 17.4 months after initial treatment. Distant disease metastasis developed in 48% of patients an average of 29 months after initial treatment. The average overall survival period was 56.2 months. In the current study, 20.6% of the probes with positive HER-2/neu expression were (+++) positive. p53 was expressed in 83.9% of the tumor samples. In 11.8% of the tumor samples, there was a lack of p16 expression. CONCLUSIONS: Mutations of the p53 gene were more frequent in tumor samples with (++) and (+++) immunoreactivity and mainly affected exons 7 and 8. A mutation of the p16 gene was only found in 1 tumor sample. Expression of HER 2/neu and p53 was statistically linked (P < 0.05) to early local disease recurrence, distant disease metastasis, and survival rates. PMID- 15900578 TI - Quantitative evaluation of telomerase subunits in urine as biomarkers for noninvasive detection of bladder cancer. AB - The aim of our study was to prospectively evaluate the potential diagnostic value and clinical applicability of quantitative analysis of telomerase subunits gene expression in urine for noninvasive detection of bladder cancer. Expression levels of human telomerase reverse transcriptase (hTERT) and human telomerase RNA (hTR) were analyzed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) in urine samples from 163 subjects with bladder cancer and 237 controls (163 individuals with benign genitourinary diseases; 74 healthy subjects). The sensitivity, specificity and optimal cutoffs were determined and compared to the corresponding values obtained by voided urine cytology. Quantitative urinary hTR analysis detects bladder cancer with an overall sensitivity of 77.0%, whereas hTERT analysis reached a sensitivity of 55.2%. The majority of undetected tumors were small, low-grade pTa lesions. Both hTR and hTERT proved to be significantly more sensitive than cytology (34.5%; p < 0.001). Specificities for hTR, hTERT and cytology were 72.1%, 85.0% and 92.7%, respectively, in the total study population and 96.9%, 89.2% and 100%, respectively, in healthy subjects. Higher diagnostic accuracy was achieved by hTR than by hTERT analysis (p < 0.05). The specificity of hTR increased to 85.0% in the total population if urinary leukocyte contamination was excluded. These data suggest that quantitative hTR analysis is the most accurate telomerase-based test for bladder cancer detection and has the potential to replace cytology as a noninvasive biomarker for disease diagnosis and follow-up. PMID- 15900579 TI - Prevalence of types 16 and 33 is increased in high-risk human papillomavirus positive women with cervical intraepithelial neoplasia grade 2 or worse. AB - High-risk human papillomavirus (hrHPV) types are causally related to cervical cancer and its high-grade precursor lesions. The risk posed by the different hrHPV types for the development of cervical intraepithelial neoplasia grade 2 or worse (> or =CIN2) needs to be established. Here, we present the hrHPV type distribution in relation to cytology and histology for women participating in a cervical screening program. From 44,102 women who participated in a population based cervical screening program in the Netherlands, 2,154 hrHPV GP5+/6+ PCR positive women were recruited to determine the distribution of 14 hrHPV types by reverse line blotting of GP5+/6+ PCR products. For each HPV type, associations with cytology and histologically confirmed > or =CIN2 were measured by odds ratios. HPV types 16 and 33 were more prevalent in women, amongst those containing a single hrHPV type, with moderate dyskaryosis or worse (>BMD) than in women with normal cytology, but only in case of underlying > or =CIN2 (OR 4.10, 95%CI 2.98-5.64 and OR 2.68, 95%CI 1.39-5.15, respectively). Similar results were obtained for women with double infections (OR 3.29, 95% CI 1.61-6.75 and OR 4.37, 95% CI 1.17-16.34). Coexisting types did not influence the prevalence of > or =CIN2 in HPV 16 or 33 positive women. The increased prevalence of type 16 and 33 in hrHPV positive women with > or =CIN2, compared to women with normal cytology, suggests that infection with these types confers an increased risk for development of > or =CIN2. Distinguishing these types may therefore have implications for future cervical screening strategies. PMID- 15900580 TI - Wnt2 as a new therapeutic target in malignant pleural mesothelioma. AB - Malignant mesothelioma of the pleura (MPM) is a highly aggressive neoplasm with a poor prognosis and limited treatment options. A better understanding of its pathogenesis is essential to developing alternative therapeutic strategies. We previously demonstrated that the Wnt signaling pathway is activated in MPM through the overexpression of disheveled proteins. To extend our knowledge of Wnt signaling activation in MPM, we performed Wnt-specific microarrays in normal pleura and MPM. We found that the most common event in MPM was the upregulation of Wnt2. We inhibited Wnt2 by siRNA and a monoclonal anti-Wnt2 antibody and analyzed their effects on apoptosis and downstream signaling effectors. We then assessed the antiproliferative effects of the Wnt2 antibody and Alimta, one of the current standard treatments of MPM. We confirmed Wnt2 overexpression at the mRNA and protein level in MPM cell lines and tissues. We then demonstrated that inhibition of Wnt2 by siRNA or a monoclonal antibody induces programmed cell death in MPM cells. We next analyzed the effects of the anti-Wnt2 antibody and of Alimta on MPM cell proliferation. We found that although Wnt2 antibody by itself had less antiproliferative potency than Alimta, the two in combination had substantially more activity than Alimta alone. We thus propose that inhibition of Wnt2 is of therapeutic interest in the development of more effective treatments for MPM. PMID- 15900581 TI - Dendritic cell maturation by CD11c- T cells and Valpha24+ natural killer T-cell activation by alpha-galactosylceramide. AB - Human invariant Valpha24+ natural killer T (NKT) cells display potent antitumor activity upon stimulation. Activation of endogenous Valpha24+ NKT cells would be one strategy for the treatment of cancer patients. For example, dendritic cells (DCs) loaded with a glycolipid NKT cell ligand, alpha-galactosylceramide (alphaGalCer, KRN7000), are a possible tool for the activation and expansion of functional Valpha24+ NKT cells in vivo. In this report, we demonstrate that the levels of expansion and the ability to produce IFN-gamma of Valpha24+ NKT cells induced by alphaGalCer-loaded whole PBMCs cultured with IL-2 and GM-CSF (IL-2/GM CSF-cultured PBMCs) were superior to those of cells induced by monocyte-derived CD11c+ DCs (moDCs) developed with IL-4 and GM-CSF. Interestingly, CD11c+ cells in the IL-2/GM-CSF-cultured PBMCs showed a mature phenotype without further stimulation and exerted potent stimulatory activity on Valpha24+ NKT cells to enable them to produce IFN-gamma preferentially at an extent equivalent to mature moDCs induced by stimulation with LPS or a cytokine cocktail. Cocultivation with CD11c- cells in the IL-2/GM-CSF-cultured PBMCs induced maturation of moDCs. In particular, CD11c-CD3+ T cells appeared to play important roles in DC maturation. In addition, TNF-alpha was preferentially produced by CD11c-CD3+ T cells in IL 2/GM-CSF-cultured PBMCs and was involved in the maturation of moDCs. Thus, the maturation of DCs induced by CD11c- T cells through TNF-alpha production appears to result in the efficient expansion and activation of Valpha24+ NKT cells to produce IFN-gamma preferentially. PMID- 15900582 TI - Lipid-lowering agents and risk of melanoma. PMID- 15900584 TI - Vitamin E amides, a new class of vitamin E analogues with enhanced proapoptotic activity. AB - Vitamin E (VE) analogues, epitomized by alpha-tocopheryl succinate (alpha-TOS), are proapoptotic agents with selective antineoplastic activity. The molecule of alpha-TOS comprises several structurally and functionally distinct moieties that can be modified in order to yield analogues with higher activity. In order to find analogues with higher apoptogenic efficacy, we prepared novel compounds where the ester bond was replaced by an amide bond. All of these analogues were significantly more proapoptotic than their ester counterparts, with alpha tocopheryl maleyl amide being the most effective. Importantly, methylation of the free carboxylic group completely obliterated apoptogenic activity of the compounds. Similarly as shown for the ester analogues, the amides induced apoptosis by mitochondrial destabilization. Superiority of amides over the ester analogues may be due to their higher partitioning into the lipid phase, as suggested by the log p-values that were lower for the amides than the corresponding esters. In conclusion, we present evidence that modification of the ester bond of agents such as alpha-TOS can be used as a basis for generating novel analogues with higher efficacy of killing malignant cells, an activity that suggests anticancer effect of the agents. PMID- 15900585 TI - Genomewide loss of heterozygosity and its clinical associations in non small cell lung cancer. AB - We extensively allelotyped a panel of 71 microdissected primary surgically resected non small cell lung cancer (NSCLC) tumors to identify chromosomal regions that are likely to contain tumor suppressor genes (TSGs) or associated with clinicopathologic and prognostic effects. Loss of heterozygosity (LOH) was detected by genotyping of 177 microsatellite markers and correlation of LOH with clinicopathologic parameters and prognosis was analyzed. Twenty markers showed an LOH frequency greater than 48%, and 8 of them (2p23.3, 2p24.3, 2q35, 6p22.2, 7p14.3, 7p22.2, 17q24.3 and 21q22.3) were novel in NSCLC. The high LOH regions were confirmed by further aligning continuous LOH regions from another set of 24 NSCLC tissues and defining 7 minimal deletion regions ranging from 1.29 to 12.26 cM. The aberrations of 8 markers showed a significant correlation with alteration of p16 and Rb proteins, suggesting the gene(s) located in the chromosomal loss that may interact with p16/Rb pathway. In addition, markers specifically associated with smoking, histology types and tumor stages were identified and the linked candidate TSGs were suggested. For example, marker D1S1612 closely linked with Mig-6 gene was associated with smoking patients, squamous cell carcinoma patients and late-stage patients. Furthermore, 3 markers, D2S2968, D6S2439 and D7S1818, were significantly associated with poor prognosis of NSCLC patients using both univariate and multivariate Cox's regression analyses (p = 0.035, 0.022 and 0.006, respectively). These markers can potentially be used for early lung cancer detection, outcome measurement and the positional cloning of new TSGs whose loss of function contributes to NSCLC tumorigenesis. PMID- 15900586 TI - Statistical inference for cancer trials with treatment switching. AB - In cancer clinical trials, it is not uncommon that some patients switched their treatments due to lack of efficacy and/or disease progression under ethical consideration. This treatment switch makes it difficult for the evaluation of the efficacy of the treatment under investigation. The current existing methods consider random treatment switch and do not take into consideration of prognosis and/or investigator's assessment that leads to patients' treatment switch. In this paper, we model patients' treatment switching effect in a latent event times model under parametric setting or a latent hazard rate model under the semi parametric proportional hazard model. Statistical inference procedures under both models are provided. A simulation study is performed to investigate the performance of the proposed methods. PMID- 15900587 TI - Estimating the proportion of cured patients in a censored sample. AB - There has been a recurring interest in modelling survival data which hypothesize subpopulations of individuals highly susceptible to some types of adverse events while other individuals are assumed to be at much less risk, like recurrence of breast cancer. A binary random effect is assumed in this article to model the susceptibility of each individual. We propose a simple multiple imputation algorithm for the analysis of censored data which combines a binary regression formulation for the probability of occurrence of an event, say recurrence of the breast cancer tumour, and a Cox's proportional hazards regression model for the time to occurrence of the event if it does. The model distinguishes the effects of the covariates on the probability of cure and on the time to recurrence of the disease. A SAS macro has been written to implement the proposed multiple imputation algorithm so that sophisticated programming effort can be rendered into a user-friendly application. Simulation results show that the estimates are reasonably efficient. The method is applied to analyse the breast cancer recurrence data. The proposed method can be modified easily to accommodate more general random effects other than the binary random effects so that the random effects not only affect the probability of occurrence of the event, but also the heterogeneity of the time to recurrence of the event among the uncured patients. PMID- 15900588 TI - A novel mouse model for segmental orthotopic colon cancer. AB - Spontaneous colon tumor mouse strains offer numerous advantages in modeling disease. However, the wide temporal window in which lesions form and the stochastic nature of lesion location require larger cohorts for assessment of disease modulation. Reliable, reproducible and inexpensive mouse models of early stage and invasive cancer would add to existing transgenic models. We show a new method for the creation of orthotopic murine tumors centered in the mucosal and submucosal layers anywhere in the colon, allowing creation of lesions of known age, location and extent. The system overcomes the disadvantages of heterotopic implantation and allows evaluation of lesions distally in the colon as well as proximally, thereby providing an additional method to study the effects of regionality. Invasion, host vascularization and application to disparate cell lines are demonstrated. Noninvasive imaging with magnetic resonance and colonoscopy, allowed in part by the tumor location, show potential applications of this approach. PMID- 15900589 TI - Omega-3 polyunsaturated fatty acids attenuate breast cancer growth through activation of a neutral sphingomyelinase-mediated pathway. AB - The effect of fish oils and their active omega-3 fatty acid constituents, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), were investigated on breast cancer growth. In in vivo experiments, mice were fed diets that were rich in either omega-3 (fish oil) or omega-6 (corn oil) fatty acids. Three weeks after implantation of MDA-MB-231 breast cancer cells, the tumor volume and weight were significantly lower (p < 0.05) for mice fed the omega-3 diets compared to those fed the omega-6 diets. Dietary fish oil also caused a 40% (p < 0.05) increase in neutral sphingomyelinase (N-SMYase) activity in the tumors. The tumor tissues from fish oil-fed animals expressed elevated p21 (waf1/cip1) mRNA, whereas tumor tissues from corn oil-fed animals exhibited undetectable levels of p21 expression. In in vitro experiments, at concentrations as low as 25 muM, DHA and EPA inhibited the growth of cultured MDA-MB-231 cells in a dose-dependent manner by 20-25% (p < 0.05). N-SMYase activity was also increased by 30-40% (p < 0.05) in the DHA- or EPA-treated cells in which an increase in ceramide formation was observed. DHA and EPA were both observed to enhance membrane bleb formation and also to induce the expression of p21. Omega-3 fatty acids-induced bleb formation and p21 expression were inhibited by the N-SMYase inhibitor GW4869, which also inhibited apoptosis by approximately 40% (p < 0.05). The results suggest that inhibition of breast cancer growth in nude mice by dietary fish oil and inhibition of breast cancer cell growth in culture by treatment with DHA and EPA is mediated by activation of N-SMYase. PMID- 15900590 TI - Mantle cell lymphomas with low levels of cyclin D1 long mRNA transcripts are highly proliferative and can be discriminated by elevated cyclin A2 and cyclin B1. AB - The role of transcript variants of cyclin D1 in cancer biology is unclear. Most tumors with high levels of cyclin D1 express 2 transcripts due to alternative splicing: one full-length transcript of 4.4 kb and one short transcript of approximately 1.7 kb. The short transcript lacks part of the 3'UTR region regulating mRNA stability and has a longer half-life. In our study, the contribution of each of these mRNAs to gene expression and cell proliferation has been investigated in mantle cell lymphoma (MCL), a B cell lymphoma characterized by a specific gene translocation resulting in enhanced expression of cyclin D1. A subset of MCL tumors with low levels of the long cyclin D1 transcript (cyclin D1 3'UTR) was identified by quantitative PCR and by oligonucleotide array hybridization. This tumor-subset had 3.4-fold higher levels of the short form of cyclin D1 mRNA (p < 0.0001) and had higher expression of cyclin D1 protein. Gene expression analysis identified a number of cell-cycle regulatory genes as upregulated. There was a significant difference in frequencies of cyclin B1 (p = 0.0006) and cyclin A2 (p = 0.0006) positive cells that discriminated MCL with low cyclin D1 3'UTR from other highly proliferative MCL. Among differentially expressed genes, there was a highly upregulated gene with homology to the group of cell-cycle promoting E2F transcription partners, E2F_TDP5. Several of the upregulated genes, such as TOP2A, AURORA A and RRM2 may influence a response to therapy. Identification of MCL with low cyclin D1 3'UTR is important because it seems to be associated with shorter overall survival. PMID- 15900591 TI - Adenoviral modulation of the tumor-associated system L amino acid transporter, LAT1, alters amino acid transport, cell growth and 4F2/CD98 expressionwith cell type specific effects in cultured hepatic cells. AB - Altered expression of metabolite transporters is observed frequently in tumor cell lines and primary neoplasms. The extent to which these may to contribute to the growth autonomy associated with cancer is not clear. LAT1 is a major L-type amino acid transporter over-expressed in a variety of cancer types and a light chain component of the CD98 heterodimer. We utilized an adenoviral expression system to modulate the level of LAT1 in a hepatic in vitro model to examine phenotypic changes associated with short-term exogenous and blocked expression. LAT1 levels were increased three fold and resulted in increased L-type amino acid transport as a result of adenoviral expression in murine hepatocytes. The protein was expressed on the cell surface and complexed with the CD98 heavy chain known as 4F2. Surprisingly, levels of the total CD98 protein complex were increased 2.4 fold as a result of adenoviral expression of light chain only, suggesting coordinate regulation. Exogenous overexpression was less effective in normal rat liver cells relative to mouse. LAT1 antisense expression in hepatic tumor cells resulted in a modest though statistically significant decrease in cell number, viability and S-phase cells over a 5-day period relative to controls despite the absence of a significant decrease in L-type transport over this period. These studies are preparatory to in vivo efforts focusing on LAT1/CD98 as a potential therapeutic target. PMID- 15900592 TI - Histologic characteristics of skin cancer in Hiroshima and Nagasaki: background incidence and radiation effects. AB - Skin cancers, though rare in Japan, have reportedly been on the rise, but little else is known about epidemiologic features of different histologic types of skin cancer. The Life Span Study cohort, which consists of 93,700 atomic-bomb survivors, many of whom were exposed to negligibly low radiation doses, and 26,600 people not exposed to radiation, enables a population-based study of spontaneous as well as radiation-related cancer risk. Skin tumor incident cases diagnosed between 1958 and 1987 were ascertained by linkage to the Hiroshima and Nagasaki tumor registries augmented by searches of other data sources. Study pathologists reviewed tumor specimens and pathology reports and classified tumors using the World Health Organization classification scheme. They identified 274 primary incident skin cancers, of which 106 were basal cell carcinoma (BCC), 81 were squamous cell carcinoma (SCC), and 14 were malignant melanomas. Background incidence rates and radiation effects were assessed by Poisson regression models allowing for the effects of demographic and other covariates. BCC and SCC background incidence rates were both about 3 per 100,000 per year. BCCs were mainly on the head/neck (81%), whereas SCCs occurred most frequently on the arms/legs (45%) and head/neck (29%), consistent with the presumed role played by solar UV exposure in skin cancer. The BCC rates increased significantly between 1958 and 1987, whereas the SCC rates remained unchanged. The excess absolute risk of BCC per unit skin surface area related to atomic-bomb radiation exposure did not differ between UV-exposed and shielded parts of the body, suggesting the additivity of the radiation-related and background BCC risks. PMID- 15900593 TI - Frequent genomic abnormalities at TWIST in human pediatric osteosarcomas. AB - The identification of genes as markers for chromosome aberrations in specific tumors might facilitate oncogenesis mechanism comprehension, cancer detection, prediction of clinical outcomes, and response to therapy. Previous physiologic and oncologic data identified the TWIST gene as a marker for mesodermal derivative and bone tissue differentiation, but its contribution to bone malignancies has not been investigated. In the present study, search for genomic alterations in high-grade pediatric osteosarcomas was focused on the 7p21 region, and more specifically on the TWIST gene. In a cohort of 74 patients, we observed by allelotyping that 31 of 68 informative tumors were rearranged at the TWIST locus. Among them, analysis by quantitative PCR (QPCR) revealed that, surprisingly, mostly deletions (22/68), but also amplifications (9/68), of the TWIST gene were detected. Furthermore, deletions at TWIST were statistically correlated to other molecular abnormalities, like alterations at the APC or c-kit loci, as well as to clinical features such as a poor outcome. This work shows that the TWIST gene seemed to be involved in high-grade pediatric osteosarcomas and is a new marker with a possible initial predictive value. PMID- 15900594 TI - Maternal height, pregnancy estriol and birth weight in reference to breast cancer risk in Boston and Shanghai. AB - Birth weight has been positively associated with breast cancer risk in adult life and is positively associated with the principal pregnancy estrogen estriol. Birth weight is lower among Chinese women than among Caucasian women, but paradoxically, pregnancy estriol levels are higher among the former than the latter. We studied a cohort of 317 Caucasian pregnant women in Boston, MA, and 339 Chinese pregnant women in Shanghai, China. We investigated whether maternal height, which is inversely associated with pregnancy estriol levels, interacts with this hormone in relation to birth weight, thus accommodating the apparently contradictory ecologic and analytic evidence concerning the role of pregnancy estrogens on breast cancer risk in the offspring. In both Boston and Shanghai, there was a positive association of pregnancy estriol with birth weight among taller women, whereas among shorter women the association was essentially null. The relevant interaction terms were highly significant in Boston (p approximately 0.006), whereas in Shanghai, where pregnant women were generally shorter, the interaction term was suggestive (p approximately 0.14). We conclude that maternal height should be considered an important risk factor for breast cancer in the offspring since it is a crucial determinant of birth weight, both directly and through positive interaction with the principal pregnancy estrogen estriol. PMID- 15900595 TI - Identification of postoperative adjuvant chemotherapy responders in non-small cell lung cancer by novel biomarker. AB - Cisplatin-based (CDDP-based) adjuvant chemotherapy of non-small cell lung cancer (NSCLC) was reported to yield 5-15% improvement in 5-year survival compared to complete resection alone. The importance of information concerning preselection of good responders has become increasingly evident. The purpose of our study is the establishment of a preselection of good responders for CDDP-based adjuvant chemotherapy. We investigated protein expressions comparing intensity between parent strains (H69 and PC14 lung cancer cultured cells) and resistant strains against CDDP using 2-dimensional polyacrylamide gel electrophoresis (2-DE). Immunohistochemically, we evaluated the relationship between protein expression associated with CDDP-resistance and the clinical effects of platinum-based postoperative adjuvant chemotherapy using 126 surgically-resected NCLC materials. We detected 2 kinds of polypeptides that changed expression levels on 2-DE gels. The analyses of the amino acid sequence showed that these polypeptides were reticulocalbin (RCN) and glutathione-S-transferase-pi (GST-pi). The 2-DE analysis showed decreased expression in RCN and overexpression in GST-pi with the acquisition of CDDP-drug resistance. RCN-transfectant of H69 CDDP-resistant strain showed intermediate sensitivity between the parent strain and the CDDP resistant strain. RCN-positive cases showed a statistically significant better disease-free survival only in the cases receiving postoperative platinum-based adjuvant chemotherapy after curative resection (p = 0.007). In addition, cases that were both RCN-positive and GST-pi-negative showed a statistically significantly better outcome (p = 0.0150). In the cases without postoperative adjuvant chemotherapy no relationship between the outcome and these expressions was seen. The evaluation of RCN and GST-pi might provide valuable information concerning postoperatively therapeutic strategy from the standpoint of individualized postoperative therapy. PMID- 15900596 TI - Akt phosphorylation associates with LOH of PTEN and leads to chemoresistance for gastric cancer. AB - Growth factor receptor-mediated signal transduction has been implicated in conferring resistance to conventional chemotherapy on cancer cells. We describe a pathway that involves AKT/PI3K to mediate chemoresistance in gastric cancer patients. Primary gastric carcinoma tissues and corresponding normal mucosa were obtained from 76 gastric cancer patients who underwent surgery in the Department of Surgery II in Kyushu University Hospital from the years 1996-2000. AKT activation was investigated by immunostaining with a phosphorylation-specific antibody, and LOH (loss of heterozygosity) of PTEN was studied in the same samples. AKT was phosphorylated in 22 cases (28.9%) of gastric cancer cases. AKT and phosphorylated AKT were not correlated with any clinicopathological factor. We found that the gastric cancer patients who had higher AKT phosphorylation (activated AKT) seemed to have LOH of PTEN (p = 0.0008). When the chemotherapeutic sensibilities of these patients were studied in an MTT assay, it was found that the activated AKT was associated with increased resistance to multiple chemotherapeutic agents (5-fluorouracil, adriamycin, mitomycin C and cis platinum). The results of our study indicate that AKT activation and LOH of PTEN plays an important role in conferring a broad-spectrum chemoresistance in gastric cancer patients. It also indicates that AKT may therefore be a novel molecular target for therapies or chemosensitivity tests that improve the outcomes of gastric cancer patients. PMID- 15900597 TI - Etiologic factors associated with p53 immunostaining in cutaneousmalignant melanoma. AB - Findings from a case-control study of cutaneous malignant melanoma (CMM) in Queensland, Australia, suggest that melanomas exhibiting p53 immunostaining possess different risk factors from those of other melanomas. To further explore this hypothesis, a case-only analysis of risk factors for p53 immunostaining with anti-p53 MAb DO-7 was undertaken in 523 people diagnosed with CMM in Canada and Australia. Phenotypic factors and past sun exposure were measured using a self administered questionnaire and telephone interview. The presence of strong p53 staining (>10% of cell nuclei positively stained vs. <1% staining) was positively associated with some indicators of high cumulative sun exposure: lentigo maligna melanoma subtype (OR = 3.2 vs. superficial spreading subtype), melanoma location on the head and neck (OR = 2.8 vs. back), histopathologic evidence of solar elastosis (OR = 2.1) and previous diagnosis of nonmelanoma skin cancer (OR = 2.4). Strong staining was negatively associated with high nevus density on the back (OR = 0.2 for >25 nevi vs. 0-3 nevi) and histologic evidence of a coexisting nevus (OR = 0.3). Other factors associated with strong p53 immunostaining include greater Breslow thickness (OR = 7.4 for >4.00 vs. <0.76 mm), male sex (OR = 2.2) and dense freckling (OR = 6.6 vs. few freckles). Of these, thickness, male sex, dense freckling, low nevus density on the back, histologic subtype and history of nonmelanoma skin cancer appeared to be independently associated with strong p53 staining. Our findings are consistent with the Queensland study in suggesting that variables indicating high accumulated sun exposure are positively associated with p53 staining and that an increased number of nevi is positively associated with its absence; they may reflect etiologic and pathogenetic heterogeneity in melanoma. PMID- 15900598 TI - Chromosome 1q loss of heterozygosity frequently occurs in sporadic insulinomas and is associated with tumor malignancy. AB - The pathogenesis of sporadic insulinomas is not clear, and there are no reliable genetic determinants that are useful to distinguish malignant and benign forms of this tumor. It was reported that 1q LOH might contribute to pathogenesis in gastrinomas and was correlated with tumor progression. However, little data are available on 1q LOH in sporadic insulinomas. In our study, we determine whether 1q LOH occurs in sporadic insulinomas and is associated with tumor malignancy by performing 1q allelotyping with 17 markers in 40 tumors and pair normal DNA. Thirty-five (88%) insulinomas had 1q LOH. Of the 35 insulinomas with 1q LOH, 14 (40%) had 1q21.3-23.2 LOH over a 7.5 cM region (SRO-1), whereas LOH in 21 tumors (60%) occurred at 1q31.3 over an 11.4 cM area (SRO-2). Of 24 tumors without MEN1 LOH, 20 had either SRO-1 or SRO-2 LOH (83%), whereas in 16 tumors with MEN1 LOH, 9 were shown to have LOH at either SRO-1 or SRO-2 (56%) (p = 0.065). This result suggests that LOH at 2 SRO might be MEN1 gene independent and may contribute to the pathogenesis in a subset of insulinomas without MEN1 gene LOH. The presence of 1q21.3-23.2 LOH is significantly associated with malignancy of insulinomas (p = 0.014). The high frequency of LOH at 1q 21.3-23.2 and 1q31.3 suggests these 2 areas may harbor putative tumor suppressor genes that may play an important role in the tumorigenesis of a subset of insulinomas. LOH at 1q21.3-23.2, which was associated with tumor malignancy, could be one of the genetic markers for identifying malignancy in sporadic insulinomas. PMID- 15900599 TI - Differential expression of steroid receptors in prostate tissues before and after radiation therapy for prostatic adenocarcinoma. AB - The expression, distribution and the role of steroid receptors in benign and malignant untreated prostate tissues is well recognized, however, the status of steroid receptors in prostate after radiotherapy (RT) for adenocarcinoma has not yet been studied fully. Immunohistochemical evaluation of androgen receptor (AR), estrogen receptor-alpha (ER-alpha), estrogen receptor-beta (ER-beta), and progesterone receptor (PR) was carried out in prostate needle biopsies obtained before and after radiotherapy from 60 patients with adenocarcinoma. The ER-beta transcripts were also studied by RT-PCR in LNCaP prostate carcinoma cell line before and 24 hr after gamma-irradiation at 0.5 Gy and 8.0 Gy. Significantly higher level of ER-beta expression was found in post-radiation samples of prostate adenocarcinoma and benign epithelium. After RT, all steroid receptors were upregulated in prostatic stroma. Tumor AR expression did not change significantly. Although a positive association between AR and ER-beta expression was observed in pre-treatment prostate adenocarcinoma, it was lost after RT suggesting that these 2 steroid receptors respond differently to RT. High levels of pretreatment tumor ER-beta were associated with local recurrence after RT and decreased biochemical recurrence-free survival (p = 0.028). LNCaP cell line that expressed no ER-beta mRNA before gamma-irradiation, clearly expressed ER-beta mRNA 24 hr after 0.5 Gy and 8.0 Gy. Upregulation of all steroid receptors in the prostate stroma and upregulation of ER-beta in the tumor epithelium after RT, may represent a protective tissue response to radiation-induced tissue injury. Although stromal AR was doubled after RT, the tumor and benign epithelium expression of AR seemed resistant to change by RT. PMID- 15900600 TI - Breast cancer risk in BRCA1 and BRCA2 mutation carriers and polyglutamine repeat length in the AIB1 gene. AB - Marked variation in phenotypic expression among BRCA1 and BRCA2 mutation carriers may be partly explained by modifier genes that influence mutation penetrance. Variation in CAG/CAA repeat lengths coding for stretches of glutamines in the C terminus of the AIB1 protein (amplified in breast cancer 1, a steroid receptor coactivator) has been proposed to modify the breast cancer risk in women carrying germline BRCA1 mutations. We genotyped the AIB1 repeat length polymorphism from the genomic DNA of a group of 851 BRCA1 and 324 BRCA2 female germline mutation carriers to estimate an association with breast cancer risk modification. Hazard ratios (HR) were calculated using a Cox proportional hazards model. For BRCA1 and BRCA2 mutation carriers, analyzed separately and together, we found that women who carried alleles with 28 or more polyglutamine repeats had no increased risk of breast cancer compared to those who carried alleles with fewer repeats (HR for BRCA1/2 carriers = 0.88, 95% CI [confidence interval] = 0.75-1.04). Analyzing average repeat lengths as a continuous variable showed no excess risk of breast cancer (BC) in BRCA1 or BRCA2 mutation carriers (HR for average repeat length in BRCA1/2 carriers = 1.01, 95% CI = 0.92-1.11). These results strongly suggest that contrary to previous studies, there is no significant effect of AIB1 genetic variation on BC risk in BRCA1 mutation carriers and provide an indication that there is also no strong risk modification in BRCA2 carriers. PMID- 15900601 TI - Androgen receptor modifications in prostate cancer cells upon long-termandrogen ablation and antiandrogen treatment. AB - To study the mechanisms whereby androgen-dependent tumors relapse in patients undergoing androgen blockade, we developed a novel progression model for prostate cancer. The PC346C cell line, established from a transurethral resection of a primary tumor, expresses wild-type (wt) androgen receptor (AR) and secretes prostate-specific antigen (PSA). Optimal proliferation of PC346C requires androgens and is inhibited by the antiandrogen hydroxyflutamide. Orthotopic injection in the dorsal-lateral prostate of castrated athymic nude mice did not produce tumors, whereas fast tumor growth occurred in sham-operated males. Three androgen-independent sublines were derived from PC346C upon long-term in vitro androgen deprivation: PC346DCC, PC346Flu1 and PC346Flu2. PC346DCC exhibited androgen-insensitive growth, which was not inhibited by flutamide. AR and PSA were detected at very low levels, coinciding with background AR activity in a reporter assay, which suggests that these cells have bypassed the AR pathway. PC346Flu1 and PC346Flu2 were derived by culture in steroid-stripped medium supplemented with hydroxyflutamide. PC346Flu1 strongly upregulated AR expression and showed 10-fold higher AR activation than the parental PC346C. PC346Flu1 proliferation was inhibited in vitro by R1881 at 0.1 nM concentration, consistent with a slower tumor growth rate in intact males than in castrated mice. PC346Flu2 carries the well-known T877A AR mutation, causing the receptor to become activated by diverse nonandrogenic ligands including hydroxyflutamide. Array based comparative genomic hybridization revealed little change between the various PC346 lines. The common alterations include gain of chromosomes 1, 7 and 8q and loss of 13q, which are frequently found in prostate cancer. In conclusion, by in vitro hormone manipulations of a unique androgen-dependent cell line expressing wtAR, we successfully reproduced common AR modifications observed in hormone-refractory prostate cancer: downregulation, overexpression and mutation. PMID- 15900602 TI - In vivo and in vitro characterization of human fibroblasts recruited selectively into human cancer stroma. AB - Fibroblasts, which are a major component of cancer-induced stroma, can have a significant impact on the progression of adjacent malignant epithelia. To characterize fibroblasts recruited into cancer-induced stroma, we examined the recruitment efficiency of 9 human fibroblast cell lines into experimental tumors generated in immunodeficient mice. Green fluorescence protein (GFP)-labeled fibroblast cell lines and human pancreatic cancer cell line Capan-1 were injected i.p. at different sites; the GFP-labeled cells within xenografts were then analyzed. KM104GFP (bone marrow) and VA-13GFP (lung) were selectively recruited into cancer stroma more efficiently than the other cell lines. KM104GFP cells did not affect tumor volume; however, VA-13GFP cells increased tumor volume by about 2-fold. After 5 cyclic in vivo passages of KM104GFP in Capan-1, we selected a subpopulation with an 8.4-fold higher recruitment efficiency (KM104GFP-5G) compared to parental KM104GFP. KM104GFP-5G also exhibited higher chemotaxis and chemoinvasion activity compared to KM104GFP in response to cancer-released chemoattractant(s). Oligonucleotide microarray analysis identified 8 genes with >3-fold upregulation and 6 genes with >3-fold downregulation in KM104GFP-5G. Immunohistochemistry confirmed that fibroblasts recruited into pancreatic cancer stroma strongly expressed carbonic anhydrase IX and keratin-8, whose transcripts were upregulated in KM104GFP-5G by oligonucleotide microarray analysis, whereas their expression in fibroblasts within noncancerous pancreatic stroma were under the detection level. Our results indicate that fibroblast recruitment is not selective with respect to organ origin and that particular fibroblast subpopulations with specific phenotypic characteristics could be recruited efficiently into cancer-induced stroma. PMID- 15900603 TI - Resection of residual disease in patients with metastatic gastrointestinal stromal tumors responding to treatment with imatinib. AB - Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors of the gastrointestinal tract. Long-term survival of patients with metastatic disease has only been observed in patients with completely resected disease. Recently, the tyrosine kinase inhibitor imatinib has been found to yield responses in the majority of patients with metastatic GIST suggesting improved resectability in responding patients. Combined treatment approaches including resective surgery after imatinib treatment in patients with advanced metastatic disease have rarely been explored. We report a series of 90 patients with metastatic GIST in whom treatment with imatinib enabled 12 patients with mostly recurrent and extensive disease to be considered for resection of residual disease. In 11 of these patients, complete resection could be achieved. Viable tumor cells were found in all but one resected specimens suggesting that despite favorable radiological or clinical responses, imatinib is unlikely to induce pathological complete responses. Until more mature data from prospective trials are available, these data suggest that an early aggressive surgical approach should be considered for all patients with metastatic GIST. Further trials investigating a combined surgical and pre/postoperative treatment with imatinib in patients with advanced metastatic GIST are warranted. PMID- 15900604 TI - International lung cancer trends by histologic type: male:female differences diminishing and adenocarcinoma rates rising. AB - Lung cancer rates have peaked among men in many areas of the world, but rates among women continue to rise. Most lung cancers are squamous cell carcinoma, small cell carcinoma, or adenocarcinoma; trends vary according to type. We compiled population-based morphology-specific incidence data from registries contributing to the International Agency for Research on Cancer (IARC) databases. Unspecified cancers and carcinomas were reallocated based on a registry, time period, sex and age group-specific basis. Where available, data from several registries within a country were pooled for analysis. Rates per 100,000 person years for 1980-1982 to 1995-1997 were age-adjusted by the direct method using the world standard. Squamous cell carcinoma rates among males declined 30% or more in North America and some European countries while changing less dramatically in other areas; small cell carcinoma rates decreased less rapidly. Squamous and small cell carcinoma rates among females generally rose, with the increases especially pronounced in the Netherlands and Norway. In contrast, adenocarcinoma rates rose among males and females in virtually all areas, with the increases among males exceeding 50% in many areas of Europe; among females, rates also rose rapidly and more than doubled in Norway, Italy and France. Rates of all lung cancer types among women and adenocarcinoma among men continue to rise despite declining cigarette use in many Western countries and shifts to filtered/low-tar cigarettes. Renewed efforts toward cessation and prevention are mandatory to curb the prevalence of cigarette smoking and to reduce lung cancer rates eventually. PMID- 15900605 TI - Generation of RAGE-1 and MAGE-9 peptide-specific cytotoxic T-lymphocyte lines for transfer in patients with renal cell carcinoma. AB - Renal cell carcinomas (RCCs) are supposed to be immunogenic, and several clinical trials of immunotherapy using tumor lysate-pulsed dendritic cells (DCs) have been performed. We report on the generation of RAGE-1 and MAGE-9 peptide-specific CTL lines. RAGE-1 and MAGE-9 are expressed in 56% and 38% of RCCs. Seven MAGE-9- and 13 RAGE-1-derived peptides were found to be immunogenic in the context of the HLA A*0201 MHC. CTLs were generated by coculture with peptide-pulsed, activated B cells, which were easily generated in great quantities and displayed functional activity for a prolonged period of time. MAGE-9 and RAGE-1 peptide-specific CTL lines were strictly peptide-specific and displayed high cytotoxic activity not only against peptide-loaded T2 cells but also against HLA-A*0201-positive RCC lines, which naturally express MAGE-9, RAGE-1 or both. Thus, B cells are well suited as APCs for the generation of large numbers of tumor peptide-specific CTLs for adoptive transfer. MAGE-9 as well as RAGE-1 may well provide suitable targets for immunotherapy of RCC. PMID- 15900606 TI - The partial area under the summary ROC curve. AB - The area under the curve (AUC) is commonly used as a summary measure of the receiver operating characteristic (ROC) curve. It indicates the overall performance of a diagnostic test in terms of its accuracy at various diagnostic thresholds used to discriminate cases and non-cases of disease. The AUC measure is also used in meta-analyses, where each component study provides an estimate of the test sensitivity and specificity. These estimates are then combined to calculate a summary ROC (SROC) curve which describes the relationship between test sensitivity and specificity across studies. The partial AUC has been proposed as an alternative measure to the full AUC. When using the partial AUC, one considers only those regions of the ROC space where data have been observed, or which correspond to clinically relevant values of test sensitivity or specificity. In this paper, we extend the idea of using the partial AUC to SROC curves in meta-analysis. Theoretical and numerical results describe the variation in the partial AUC and its standard error as a function of the degree of inter study heterogeneity and of the extent of truncation applied to the ROC space. A scaled partial area measure is also proposed to restore the property that the summary measure should range from 0 to 1. The results suggest several disadvantages of the partial AUC measures. In contrast to earlier findings with the full AUC, the partial AUC is rather sensitive to heterogeneity. Comparisons between tests are more difficult, especially if an empirical truncation process is used. Finally, the partial area lacks a useful symmetry property enjoyed by the full AUC. Although the partial AUC may sometimes have clinical appeal, on balance the use of the full AUC is preferred. PMID- 15900607 TI - Total loss of MHC class I is an independent indicator of good prognosis in breast cancer. AB - Tumours can be recognised by CTL and NK cells. CTL recognition depends on expression of MHC Class I loaded with peptides from tumour antigens. In contrast, loss of MHC Class I results in NK activation. In our study a large set of samples from patients with primary operable invasive breast cancer was evaluated for the expression of MHC Class I heavy and light by immunohistochemical staining of 439 breast carcinomas in a tissue microarray. Forty-seven percent (206 of 439) of breast carcinomas were considered negative for HLA Class I heavy chain (HC10), whereas lack of anti-beta2m-antibody staining was observed in 39% (167 of 424) of tumours, with only 3% of the beta2m-negative tumours expressing detectable HLA Class I heavy chain. Correlation with patient outcome showed direct relationship between patient survival and HLA-negative phenotype (log rank = 0.004). A positive relationship was found between the intensity of expression of MHC Class I light and heavy chains expression and histological grade of invasive tumour (p < 0.001) and Nottingham Prognostic Index (p < 0.001). To investigate whether HLA Class I heavy and light chains expression had independent prognostic significance, Cox multivariate regression analysis, including the parameters of tumour size, lymph node stage, grade and intensity of HC10 and anti-beta2m staining, was carried out. In our analysis, lymph node stage (p < 0.001), tumour grade (p = 0.005) and intensity of MHC Class I light and heavy chains expression were shown to be independent prognostic factors predictive of overall survival (p values HC10 = 0.047 and beta2m = 0.018). PMID- 15900608 TI - Novel functional biodegradable polymer. III. The construction of poly(gamma glutamic acid)-sulfonate hydrogel with fibroblast growth factor-2 activity. AB - Poly(gamma-glutamic acid)-sulfonate (gamma-PGA-S) hydrogel, with fibroblast growth factor (FGF)-2 activity was investigated as a novel, next-generation tissue-engineering material. gamma-PGA-net-gamma-PGA-S72 (S72-netgel) was prepared with gamma-PGA-S and gamma-PGA-S72 (72% sulfonated gamma-PGA) to provide the high mobility of gamma-PGA-S72 for FGF-2 activity. Cell adhesion and proliferation activities were evaluated on gamma-PGA and gamma-PGA-S hydrogels along with S72-netgels. Both cell adhesion and proliferation activities of gamma PGA and gamma-PGA-S hydrogels were low. In contrast, S72-netgels had high cell adhesion and proliferation activities, because of their low swelling ratios and high sulfonic acid group concentrations. Furthermore, S72-netgels had high FGF-2 activity, because gamma-PGA-S72 retained FGF-2 activity when incorporated into S72-netgels. S72-netgels should be useful as next-generation tissue-engineering material containing FGF-2 activity. PMID- 15900609 TI - Evaluation of modifying collagen matrix with RGD peptide through periodate oxidation. AB - The aim of the study is to evaluate the effect of modifying collagen matrices with Arg-Gly-Asp (RGD) peptide through periodate oxidation. The collagen matrices were modified with RGD peptide, by periodate activation. The modified collagen matrices and unmodified matrices were characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and electron spectroscopy for chemical analysis (ESCA). Mesenchymal stem cells (MSCs) were used to evaluate the cell compatibility of collagen matrices. In terms of cell growth, the MSCs attached much better on the modified matrix than on the unmodified one. But there was no significant difference between two groups regarding the MSC proliferation. Compared to the unmodified matrices, the mechanical strength of the modified matrix decreased sharply, and its 3D structure was destroyed. Introducing specific RGD receptor-mediated adhesion sites on matrices obviously enhanced the MSC adhesion on collagen matrices, but the coupled method of periodate oxidation would likely result in the declination of the mechanical strength of the matrix, as well as the destruction of the matrix structure. This would affect the cell growth on the matrix, and decrease the histocompatibility of the matrices. PMID- 15900610 TI - Effect of synovial fluid, phosphate-buffered saline solution, and water on the dissolution and corrosion properties of CoCrMo alloys as used in orthopedic implants. AB - The corrosion and dissolution of high- and low-carbon CoCrMo alloys, as used in orthopedic joint replacements, were studied by immersing samples in phosphate buffered saline (PBS), water, and synovial fluid at 37 degrees C for up to 35 days. Bulk properties were analyzed with a fine ion beam microscope. Surface analyses by X-ray photoelectron spectroscopy and Auger electron spectroscopy showed surprisingly that synovial fluid produced a thin oxide/hydroxide layer. Release of ions into solution from the alloy also followed an unexpected pattern where synovial fluid, of all the samples, had the highest Cr concentration but the lowest Co concentration. The presence of carbide inclusions in the alloy did not affect the corrosion or the dissolution mechanisms, although the carbides were a significant feature on the metal surface. Only one mechanism was recognized as controlling the thickness of the oxide/hydroxide interface. The analysis of the dissolved metal showed two mechanisms at work: (1) a protein film caused ligand-induced dissolution, increasing the Cr concentration in synovial fluid, and was explained by the equilibrium constants; (2) corrosion at the interface increased the Co in PBS. The effect of prepassivating the samples (ASTM F-86-01) did not always have the desired effect of reducing dissolution. The release of Cr into PBS increased after prepassivation. The metal-synovial fluid interface did not contain calcium phosphate as a deposit, typically found where samples are exposed to calcium rich bodily fluids. PMID- 15900611 TI - Expression pattern of the chromosome 21 transcription factor Ets2 in cell-seeded three-dimensional bone constructs. AB - The ability to generate new bone for reconstructive surgery use is a major clinical need. Tissue engineering with osteoprogenitor cells isolated from the patient's periosteum and seeded into bioresorbable scaffolds offers a promising approach to the generation of skeletal tissue. To our knowledge, there is no description about the expression of Ets2 in tissue engineered "bone neotissue." The aim of our study was to manufacture cell-seeded three-dimensional bone constructs with human periosteal cells on poly (lactic-co-glycolic acid) polymer fleeces to describe the expression pattern of Ets2 and its target genes osteocalcin and osteopontin; expression analysis of type I collagen, core-binding factor-1, alkaline phosphatase, and osteonectin; the ability of matrix mineralization and ALP enzymatic activity showed the osteogenic character of the constructs. A significant correlation between the expression of Ets2 and osteopontin mRNA (r = -0.70; p < 0.05) could be shown. A 1.35-fold increase of Ets2 expression from days 1 to 9 was detected, followed by a slight decrease from days 11 to 15. Until the end of the culture period, the expression of Ets2 reached a comparable high level as detected on day 9. In contrast, the expression level of osteopontin mRNA reached a maximum at day 7, followed by a progressive 3.04-fold decrease until day 21. This study shows for the first time that Ets2 gene and its transcriptional target genes are expressed in tissue-engineered bone constructs. These findings have the potential to provide much-needed information about the role and function of Ets2 in human osteogenesis processes and creation of "bone neotissue." PMID- 15900614 TI - Development of new injectable bulking agents: biocompatibility of radiopaque polymeric microspheres studied in a mouse model. AB - Radiopaque polymeric microspheres have a potential as new bulking agents for treatment of stress urinary incontinence (SUI). The advantage over existing bulking agents lies in their X-ray visibility in situ; other polymeric bulking agents (e.g., PTFE or silicone rubbers) are practically radiolucent (i.e., incapable of absorbing X-radiation). Radiopacity is useful in practice because of the high spatial accuracy of X-ray imaging. For instance, X-ray fluoroscopy can be used to assess possible migration of the bulking agent over time or to provide guidance in cases in which a second injection of a bulking agent is necessary (repeated treatment of SUI). Biocompatibility of injected radiopaque microspheres was investigated in vivo by using the mouse as a model. Microspheres were injected subcutaneously (9 animals) or intramuscularly (9 animals), and follow-up was 8 days or 3 months. X-ray fluoroscopy gave clear images of the microspheres as an ensemble, and it was found that no migration occurred during 3 months. Histopathology confirmed that all microspheres stayed close to the site of the injection. The microspheres appeared to be well tolerated; only a few giant cells, manifesting a mild inflammatory reaction, were encountered. At 3 months, capillary blood vessels were observed throughout the microsphere beds, and macrophages and fibroblast cells were seen in between the microspheres. This is encouraging with respect to the intended application, although it must be acknowledged that the data refer merely to a mouse model. Further experiments with larger, more representative models (rabbit and goat) are in progress. PMID- 15900615 TI - Fabrication, implantation, elution, and retrieval of a steroid-loaded polycaprolactone subretinal implant. AB - A subretinal drug delivery system was developed to overcome the limitations of current treatments for retinal disease. A rod-shaped implant was made by embedding the corticosteroid triamcinolone acetonide within a biodegradable polycaprolactone polymer matrix. The implant was fabricated by homogeneously mixing the polymer and drug in solvent. The mixture was then dried, melted, and extruded, and the prepared solid form was drawn into a filament. The rods were mechanically sectioned to a length of 2 mm with a diameter of up to 320 microm. The rods were successfully implanted into the subretinal space of six rabbits. No complications were observed during the 4-week follow-up period. Initial observations of the implantation and elution characteristics revealed that polycaprolactone is well tolerated by the retinal tissue and that the implant can elute steroid for a period of at least 4 weeks without eliciting inflammatory response or complications. In vitro drug elution rates of different polymer to drug ratios and geometries into a balanced salt solution/bovine serum albumin (1%) solution showed an early rapid-release phase and late first-order phase. Histology and device retrieval after implantation revealed minimal encapsulation and good preservation of cellular morphology during the follow-up period and a more fibrous polymer microstructure of the implant. PMID- 15900616 TI - Liquid chromatography-electron ionization mass spectrometry: fields of application and evaluation of the performance of a Direct-EI interface. AB - A comprehensive evaluation and a thorough discussion of the fields of possible applications of the Direct-EI interface are described in this review. Direct-EI allows the direct introduction of the effluent from a capillary HPLC column into the electron ionization (EI) ion source of a mass spectrometer. Thanks to the reduced liquid intake and an in-source nebulizer, the interfacing process occurs smoothly and entirely into the ion source. No intermediate interfacing mechanisms of any sort are interposed between the column and the mass spectrometer, thus circumventing any undesired sample loss and minimizing the effort for instrument modification. Theoretically, any GC-MS system can be converted into an LC-MS for EI amenable compounds. Several parameters, crucial for a successful integration of liquid chromatography and mass spectrometry, have been considered in the evaluation of the functioning of such an interface: limit of detection, linearity of response, reproducibility, and chromatographic compatibility. Different mobile phases, also containing non-volatile buffers, were taken into account, demonstrating an outstanding separation flexibility. The entire set of experiments was carried out at different flow rates and temperatures of the ion source. The interface behavior was also tested in real world applications, with mixtures of pesticides, hormones, nitro-PAH, and endocrine-disrupting compounds, allowing picogram level detection and the possibility to record library matchable, readily interpretable electron ionization mass spectra, for prompt compound characterization and confirmation. PMID- 15900617 TI - Pharmacokinetics and tissue distribution of d-alpha-tocopheryl succinate formulations following intravenous administration in the rat. AB - Unlike d-alpha tocopherol (T), d-alpha tocopheryl succinate (TS) has the unique ability to selectively kill tumor cells while protecting normal tissue from toxic oxidative stress. The pharmacokinetics of TS and the serum and tissue disposition of TS were studied in male Sprague-Dawley rats to delineate formulation dependent disposition between TS administered as the Tris salt (TS-T) (a liposomal formulation) or as the free acid (TS-FA) dissolved in polyethylene glycol (PEG) 400. The pharmacokinetics of TS was studied after single intravenous (i.v.) equimolar doses of 124 mg/kg TS-T and 100 mg/kg of TS-FA. Serial blood samples were collected via a catheter inserted into the right jugular vein and serum samples were analysed for TS and T levels using a reverse phase HPLC method. Terminal tissue samples were also collected at 24 and 48 h. After an acute i.v. dose of TS-T, serum AUC, t(1/2), Cl and V(d) of TS were 2601.0 +/- 351.7 microg h/ml, 9.98 +/- 1.02 h, 0.049 +/- 0. 0073 l/h/kg and 0.7 +/- 0.14 l/kg (mean +/- SD), respectively. The acute i.v. administration of TS-FA (PEG formulation) yielded results similar to those observed for TS-T, with a serum AUC, t(1/2), Cl and V(d) of 2553.3 +/- 166.4 microg h/ml, 9.83 +/- 0.86 h, 0.039 +/- 0.0027 l/h/kg and 0.56 +/- 0.09 l/kg (mean +/- SD), respectively. Distribution into tissues and a low Cl was apparent, with the highest concentrations of TS in the liver and lung, regardless of formulation. As expected, baseline endogenous concentrations of T were present in both groups, with a net increase in T levels, occurring as TS was hydrolysed to T, which slowly peaked in serum between 7-8 h post-dose. Intravenous TS administration, regardless of formulation, also resulted in significant T accumulation in all tissues examined, which was especially abundant in the liver and lung. Likewise, there was a lack of significant effect of formulation on the pharmacokinetics and tissue distribution of TS. The only significant formulation difference was a small but significant increase in serum T and liver T levels in the TS in PEG formulation group. These results indicate that TS may be especially useful for the targeted delivery of T and TS to the lung and liver for anti-oxidant and anti-cancer activity. PMID- 15900618 TI - Perspectives. What to expect in Medicare Rx: varying premiums, PDPs for duals. PMID- 15900619 TI - Electronic "eye" aids heart failure. PMID- 15900620 TI - Ask the doctor. I am 86 and have atrial fibrillation. I started taking warfarin to avoid blood clots, but right away developed a very high bleeding time and needed to be hospitalized. I don't want to go through that again. Is there another approach for me? PMID- 15900621 TI - Risks associated with blood transfusion after total knee arthroplasty. PMID- 15900622 TI - Letters regarding article by Bloomfield et al, "microvolt T-wave alternans distinguishes between patients likely and patients not likely to benefit from implanted cardiac defibrillator therapy". PMID- 15900623 TI - Should everyone with an ejection fraction less than or equal to 30% receive an implantable cardioverter-defibrillator? Everyone with an ejection fraction < or = 30% should receive an implantable cardioverter-defibrillator. PMID- 15900624 TI - Gastrointestinal perforation and cancer therapy: managing risk to achieve benefit. PMID- 15900625 TI - Erythropoietin in the treatment of cancer. PMID- 15900626 TI - Management of vancomycin-resistant enterococci--a paradigm. PMID- 15900627 TI - Successful treatment of Naegleria fowleri meningoencephalitis by using intravenous amphotericin B, fluconazole and rifampicin. AB - BACKGROUND: Primary amebic meningoencephalitis (PAM) is an emerging disease with a rapidly fatal outcome. Only eight reports of cured cases have appeared in the medical literature to date. METHODS: A 10-year-old boy developed PAM caused by Naegleria fowleri 1 week after swimming in an irrigation canal. He was admitted to our hospital after 9 h of severe headache and vomiting, fever, ataxic gait, mild confusion, and seizures were evident. Trophozoites were identified in the cerebrospinal fluid (CSF). Treatment with intravenous (i.v.) dexamethasone, amphotericin B, fluconaloze, and oral rifampicin was started. After several hours of conflicting clinical signs, recovery began, and on the third day he was conscious again. Hospital discharge occurred on day 23, after a normal brain CT scan. There was no sequel to the disease during the following 12 months. RESULTS: The amebas present in the CSF were identified and confirmed as N. fowleri after observation of wet mounts and of cultures seeded on 1.5% non-nutrient agar plates covered with Escherichia coli, vegetative and cystic forms, enflagellation experiments in distilled water at 98 degrees F, temperature tolerance testing and by indirect immunofluorescence using N. fowleri LEE antibody. The genotype was determined by PCR amplification and sequencing of the internal transcribed spacers (ITS) including the 5.8S rDNA. CONCLUSIONS: Early treatment of PAM by i.v. administration of amphotericin B and fluconazole, and oral administration of rifampicin can offer some hope of cure for this devastating disease. PMID- 15900628 TI - The role of plasma high molecular weight kininogen in experimental intestinal and systemic inflammation. AB - Inflammation is accompanied by activation of the plasma kallikrein-kinin system (KKS). KKS activation has been demonstrated in a variety of inflammatory human diseases. To further explore the participation of KKS in arthritis and inflammatory bowel disease, we used two experimental animal models in arthritis and enterocolitis. We found that activation of KKS is associated with arthritis induced by intraperitoneal injection of peptidoglycan-polysaccharide polymers (PG PS) as well as the enterocolitis and systemic inflammation induced also by PG-PS when injected into the intestinal wall of genetically susceptible Lewis rats. We postulated that KKS participates in the pathogenesis of inflammatory reactions involved in cellular injury, coagulation, fibrinolysis, kinin formation, complement activation, cytokine secretion, and release of proteases. We demonstrated that therapy with a specific plasma kallikrein inhibitor modulated the experimental enterocolitis, arthritis, and systemic inflammation. The fact that deficiency of plasma high molecular weight kininogen in the genetically susceptible Lewis rat results in decreased chronic enterocolitis and systemic inflammation also supports our hypothesis. We suggest that KKS plays a similar role in idiopathic human intestinal inflammatory disease and arthritis, making kallikrein-kinin system proteins appealing targets for drug therapy in chronic inflammatory diseases such as rheumatoid arthritis and Crohn's disease. PMID- 15900629 TI - HIV immune pathogenesis: an update from CROI. PMID- 15900630 TI - HIV and insulin resistance in context. PMID- 15900631 TI - Policy watch. Will the new women get it? PMID- 15900632 TI - HIV-hepatitis B virus coinfection: questions and answers. PMID- 15900633 TI - Recent developments in HIV treatment, Part 1. PMID- 15900634 TI - Primary HIV-1 infection: to treat or not to treat? AB - Treatment of primary HIV-1 infection with HAART may represent a unique opportunity to alter the course of this chronic disease. In spite of this, enthusiasm for the "ht early, hit hard" approach has waned over time, given the lack of data showing long-term clinical benefit. This article will examine the risks and benefits of starting HAART during acute HIV-1 disease. PMID- 15900635 TI - Editorial comment: treatment of acute HIV infection--uncertainties about best practice. PMID- 15900636 TI - Cryoglobulinemic vasculitis and lymphocytic interstitial pneumonitis in a person with HIV infection. AB - A wide variety of rheumatologic complications have been reported in the setting of HIV-1 infection. We present the case of an HIV-1-infected patient with small vessel vasculitis and inflammatory lung disease. This case illustrates the complexity of the differential diagnosis of inflammatory and autoimmune disease states in HIV-1-infected patients. PMID- 15900637 TI - Effect of carbon ions on life span shortening and tumorigenesis in mice. AB - One of the important concerns for astronauts in space is cancer risk associated with cosmic radiation, including heavy particle ions. But little information on cancer risk is available. We investigated the effect of carbon ions on life span shortening and tumor induction in B6C3F1 mice. The mice were exposed weekly to 0.4 and 2.0 Gy whole-body carbon-ion- or X-ray-irradiation for 4 consecutive weeks. The spectrum of induced tumors varied depending on the dose. The cause of death was thymic lymphomas and liver tumors at high and low dose, respectively. The life span shortening by X-rays was proportional to dose, while carbon ions produced a convex upward relationship. The relative biological effectiveness for the 50% life span shortening was about 1.4. The large effect of carbon ions encourages the study on tumor induction at low doses in the space. PMID- 15900639 TI - Heart Rhythm 2004. Abstracts of the 25th Annual Meeting of the Heart Rhythm Society. May 19-22, 2004, San Francisco, California, USA. PMID- 15900638 TI - Abstracts of the 149th meeting of the Acoustical Society of America joint meeting with the Canadian Acoustical Association. Vancouver, Canada, 16-20 May 2005. PMID- 15900640 TI - Digestive Disease Week and the 106th Annual Meeting of the American Gastroenterological Association. May 14-19, 2005, Chicago, Illinois, USA. Abstracts. PMID- 15900641 TI - A career in nursing. PMID- 15900642 TI - Fluids containing a highly branched cyclic dextrin influence the gastric emptying rate. AB - The rates of gastric emptying for highly branched cyclic dextrin (HBCD) and other carbohydrate (CHO) solutions were examined using ultrasonograph techniques. Ten healthy volunteers ingested water, physiological saline, or solutions containing various CHO, such as HBCD, glucose, maltose, sucrose, and commercially available dextrin. After a subject drank one of the solutions, the relaxed cross-sectional area of the pylorus antrum was measured at rest by real-time ultrasonography. The time required for gastric emptying was correlated with the relaxed cross sectional area of the pylorus antrum. Among all of the solutions tested, physiological saline was transferred fastest from the stomach to the small intestine. For solutions of the same CHO, 5 % solution was transferred faster than 10 % solution. For CHO solutions other than HBCD, a low osmotic pressure was associated with rapid transfer from the stomach. The gastric emptying time (GET) of HBCD solution increased with an increase in its concentration. A shorter GET was observed for the CHO solutions at 59 to 160 mOsm regardless of their concentration. A sports drink based on 10 % HBCD adjusted to 150 mOsm by the addition of various minerals, vitamins, and organic acids was evacuated significantly (p < 0.05) faster than a 10 % HBCD solution or a sports drink based on 10 % commercially available dextrin (DE16), which has a higher osmotic pressure (269 mOsm). Our results suggest that a shorter GET could be achieved with CHO solutions with osmotic pressures of 59 - 160 mOsm. Therefore, a sports drink based on 10 % HBCD adjusted to 150 mOsm by the addition of minerals, vitamins, and organic acids could supply adequate quantities of CHO, fluid, and minerals simultaneously in a short time, without increasing GET. PMID- 15900643 TI - [In memory of Iurii Aleksandrovich Ovchinnikov]. PMID- 15900644 TI - The fluid-dynamic disturbances induced on the ISS, based on the first acceleration measurements on board the space station. AB - The different acceleration components on the ISS that are responsible for the generation of convective motions in a fluid cell either in the presence of density gradients or in quasi-isodense processes, are analyzed. The NASA measurements of the quasi-steady and periodic acceleration on the ISS are considered and their effects on fluid-dynamic experiments are computed and discussed under different assumptions. In particular, numerical simulations are carried out to identify the relative importance of linear and pendular accelerations, due to possible rotations of the P/L around its center of mass. The effects caused by variable accelerations created by an isolation mount that exhibits an attenuation factor not constant within the payload volume, caused by the reaction forces of the umbilicals, are computed and analyzed. PMID- 15900645 TI - Immobilization induces a very rapid increase in osteoclast activity. AB - We studied in a randomized, strictly controlled cross-over design, the effects of 6 days 6 degrees head-down tilt bed rest (HDT) in eight male healthy subjects in our metabolic ward. The study consisted of two periods (phases) of 11 days each in order to allow for the test subjects being their own controls. Both study phases were identical with respect to environmental conditions, study protocol and diet. Two days before arriving in the metabolic ward the subjects started with a diet. The diet was continued in the metabolic ward. The metabolic ward period (1l days) was divided into three parts: 4 ambulatory days, 6 days either HDT or control and 1 recovery day. Continuous urine collection started on the first day in the metabolic ward to analyze calcium excretion and bone resorption markers. On the 2nd ambulatory day in the metabolic ward and on the 5th day in HDT or control blood was drawn to analyze serum calcium, parathyroid hormone, and bone formation markers. Urinary calcium excretion was, as early as the first day in immobilization, increased (p<0.01). CTX- and NTX-excretion stayed unchanged in the first 24 h in HDT compared to the control. But already on the 2nd day of immobilization, both bone resorption markers significantly increased. We conclude from these results--pronounced rise of bone resorption markers--that already 24 h of immobilization induce a significant rise in osteoclast activity in healthy subjects. Thus, it appears possible to use short-term bed rest studies as a first step for the development of countermeasures to immobilization. PMID- 15900646 TI - Mars habitat modules: launch, scaling and functional design considerations. AB - The Sasakawa International Center for Space Architecture (SICSA) is undertaking a multi-year research, planning and design study that is exploring near- and long term commercial space development opportunities. The central goal of this activity is to conceptualize a scenario of sequential, integrated private enterprise initiatives that can carry humankind forward to Mars. Each development stage is planned as a building block to provide the economic foundation, technology advancements and operational infrastructure to support others that follow. This report presents fundamental issues and requirements associated with planning human Mars initiatives that can transfer crews, habitats and equipment from Earth to Mars orbit, deliver them to the planet's surface, and return people and samples safely back to Earth. The study builds in part upon previous studies which are summarized in SICSA's: Commercial Space Development Plan and the Artificial Gravity Science and Excursion Vehicle reports. Information and conclusions produced in this study provide assumptions and a conceptual foundation for a subsequent report titled The First Mars Outpost: Planning and Concepts. PMID- 15900647 TI - Home health payment reform: trends in the supply of rural agencies and availability of home-based skilled services. PMID- 15900648 TI - [Analgesics in pregnancy]. PMID- 15900649 TI - Competition and disclosure incentives: an empirical study of HMOs. AB - I examine Health Maintenance Organizations' (HMOs) voluntary disclosure of product quality, which is not as complete as unraveling theories predict. After controlling for cost and demand factors, I find that HMOs use voluntary disclosure to differentiate from competitors, with lower disclosure rates in highly competitive markets. These findings are consistent with product differentiation, but challenge the intuition that competition should lead to more provision of quality information. PMID- 15900650 TI - Postpartum ovarian vein thrombosis. PMID- 15900651 TI - High prevalence of 5G allele in Amerindian tribes and Mestizos from Mexico at 4G/5G PAI-I gene promoter polymorphism. PMID- 15900652 TI - The Activated Seven Lupus Anticoagulant (ASLA) test has comparable sensitivity to classical assays for screening of lupus anticoagulant. PMID- 15900653 TI - Hospital oversight in Medicare: accreditation and deeming authority. AB - To be eligible to receive payment from Medicare, hospitals must be certified to meet certain conditions. Hospitals may gain such credentials by choosing to be reviewed by a state certification agency under contract to the Centers for Medicare & Medicaid Services or to be accredited by either the Joint Commission on Accreditation of Healthcare Organizations or the American Osteopathic Association. This issue brief looks at how accreditation of hospitals developed and how it continues to change. It considers the legal and practical reasons that a majority of hospitals choose accreditation and why some hospitals do not, along with broader consideration of the extent to which accreditation may be judged of value to Medicare beneficiaries. The intersection of state and federal oversight responsibilities and the role of accrediting organizations in hospital quality improvement also are examined. PMID- 15900654 TI - An update on Americans' access to prescription drugs. AB - More Americans--especially those with chronic conditions such as diabetes, asthma and depression--are going without prescription drugs because of cost concerns, according to a new national study by the Center for Studying Health System Change (HSC). In 2003, more than 14 million American adults with chronic conditions- over half of whom were low income--could not afford all of their prescriptions. Between 2001 and 2003, the proportion of privately insured, working-age people with chronic conditions who reported not filling at least one prescription because of cost concerns increased from 12.7 percent to 15.2 percent. Likewise, the proportion of elderly, chronically ill Medicare beneficiaries without supplemental private insurance with problems affording prescription drugs rose from 12.4 percent to 16.4 percent between 2001 and 2003. At the same time, significant disparities in prescription drug access persisted between black and white Americans with chronic conditions, with blacks about twice as likely to report problems affording prescriptions. PMID- 15900655 TI - The need for new therapeutic agents: what is the pipeline? AB - There is a clinical need for new treatment options for serious Gram-positive infections. Recently introduced agents such as the newer fluoroquinolones and the ketolide telithromycin have limited use as they do not cover methicillin resistant Staphylococcus aureus (MRSA) or glycopeptide-resistant enterococci (GRE). The clinical use of the streptogramin combination quinupristin/dalfopristin, which has activity against MRSA and vancomycin resistant Enterococcus faecium, is limited because administration is via a slow infusion of a large volume. The oxazolidinone linezolid is active against MRSA and GRE but resistant organisms and treatment failures have been reported. A number of compounds currently in development show promise, the new glycopeptides oritavancin, dalbavancin and the glycolipodepsipeptide ramoplanin, as well as the new tetracyclines tigecycline and BAY73-7388. However, in some cases, there is concern that resistance may develop quickly to new compounds that are based on existing antimicrobial agents. Therefore daptomycin, a novel lipopeptide with a unique mode of action, is of particular interest. Daptomycin is active against MRSA (including vancomycin-resistant strains) and GRE. Daptomycin displays rapid concentration-dependent killing and is bactericidal even in the stationary phase of growth. Daptomycin-resistant strains are very difficult to generate in vitro. A dosage of 4 mg/kg intravenous once a day has been shown to be efficacious in two evaluator-blinded trials of complicated skin and soft tissue infections with clinical success rates similar for daptomycin and comparators (vancomycin or penicillinase-resistant penicillins). With its activity against key Gram-positive pathogens, including resistant strains, daptomycin has potential as a valuable addition to the available treatment options for serious Gram-positive infections. PMID- 15900656 TI - Holistic nursing theory: connecting heart & mind in practice. PMID- 15900657 TI - The language of holism. PMID- 15900658 TI - Attaining legitimacy with holistic nursing research. PMID- 15900659 TI - One thing I know for sure about poetry. PMID- 15900660 TI - Innovative communication brings holism to Duke. PMID- 15900661 TI - Interested in policy? Start with your state BON. Interview with Geneie Everett. PMID- 15900662 TI - Degradability and sediment sorption of an alcohol polyglycol ether surfactant putatively useful for the control of red swamp crayfish in rice fields. AB - This work reports studies of the degradation rates of a fatty alcohol polyglycol ether non-ionic surfactant, Genapol OXD-080, putatively useful for the control of red swamp crayfish (Procambarus clarkii Girard) in rice fields under laboratory and field conditions. The influence of temperature, sediment site specificity and sorption were taken into account. The degradation kinetics of the surfactant depends on the experimental conditions: type of inocula and temperature. The distribution of this chemical in aquatic systems was also examined. Genapol OXD 080 was removed into the sediments readily after application, and sorption was considered the major path of removal from the water phase. Data suggest that further studies are required regarding the effects of Genapol OXD-080 in aquatic organisms resident in rice fields, in parallel with the development of technologies related with the use of surfactants to control P. clarkii populations. PMID- 15900663 TI - Environmental impact assessment of the mining and concentration activities in the Kola Peninsula, Russia by multidate remote sensing. AB - On the Kola Peninsula, the mining and concentration industry exerts anthropogenic impact on the environment. Tailing dumps cause airborne pollution through dusting, and waterborne pollution due to direct dumping and accidental releases. The objectives were: (1) to analyse multidate satellite images for 1964-1996 to assess the environmental pollution from the mining and concentration activity in the Kola in temporal perspective; (2) to evaluate remote sensing methods for integrated environmental impact assessment. The area of mining and industrial sites steadily expands and amounted to 94 km2 in 1996. The polluted water surface amounted to at least 150 km2 through dumping in 1978 and to 106 km2 in 1986 due to dusting. Thus, the impact from the mining and concentration activity should be reconsidered as more significant than it was officially anticipated. In the past the main mechanism of pollution was direct dumping into the lakes. Currently and in future, airborne pollution after dusting storms will dominate. The effective recultivation of the tailing dumps will be a long-term process. For effective assessment of impacts from the mining and concentration industry, remote sensing methods should be complemented by in-situ measurements, fieldwork, and mathematical modelling. PMID- 15900664 TI - Volatile organic compounds in the area of Madrid: a chemometrical approach. AB - Multivariate techniques have been applied to the set of data obtained after one year sampling of volatile organic compounds (VOCs), including volatile aldehydes in the area of Madrid in order to evaluate the possible correspondence among groups of analysed compounds, as far as modelling of different emission sources in relation to location and season. The measurements were carried out at four sites in Madrid, characteristic for urban and suburban areas. Additionally, as reference, a rural area 100 km far from the city was considered. Results of correlation analysis, factor and cluster analysis are presented. Higher correlations were found between variables related with traffic emissions. Factor analysis results showed two main significant variables related to anthropogenic and biogenic emissions respectively. In relation to cluster analysis, samples were grouped according to sampling site and seasonal variations. PMID- 15900665 TI - Evaluation of metric precision for a riparian forest survey. AB - This article evaluates the performance of a protocol to monitor riparian forests in western Oregon, United States based on the quality of the data obtained from a field survey. Precision is the criteria used to determine the quality of 19 field and 6 derived metrics. The derived metrics were calculated from the field data. The survey consisted of 110 riparian sites on public and private lands that were sampled during the summers of 1996 and 1997. In order to calculate metric precision, some of the field plots were re-measured. Metric precision was defined in terms of the coefficient of variability (CV) and standard deviation and then compared with a pre-defined data quality objective (DQO). A metric was considered precise if the CV met or exceeded the DQO. The geomorphology metrics were not precise while the forest stand inventory metrics and forest cover metrics, with some exceptions, were precise. The precision for many of the field and derived metrics compared favorably with the level of precision for similar metrics reported in the literature. Recommendations are made to improve the precision for some metrics and they include changing the way precision is calculated, re defining the field protocol, or improving field training. PMID- 15900666 TI - Assessment of calibration parameters for an aerial gamma spectrometry system using Monte-Carlo technique. AB - During a radiation emergency subsequent to a nuclear accident or weapon fallout, quick assessment of the ground contamination and the resulting exposure is of prime importance in planning and execution of effective counter measures. For an online assessment of ground contamination, it is essential to calibrate the detector system for several parameters viz. the source energy, source deployment matrix, the flight altitude and position above the contaminated surface. This article discusses the methodology to predict all the necessary parameters like photon fluence at various altitudes, the photo-peak counts in different energy windows, Air to Ground Correlation Factors IAGCF) and the dose rate at any height due to air scattered gamma ray photons. The methodology includes generation of theoretically simulated gamma spectra at a required detector position for a given source distribution on the ground using Monte-Carlo method provided by general purpose coupled neutron/photon transport code (MCNP CCC-200). Thus generated gamma spectra are analyzed to arrive at the required parameters mentioned above. PMID- 15900667 TI - Rate-limited desorption of volatile organic compounds from soils and implications for the remediation of a Louisiana Superfund site. AB - The rates of desorption of trichloroethylene (TCE) and 1,3-dichlorobenzene (DCB) from a silty soil at a Superfund site and a silty-clayey soil from an uncontaminated bottomland hardwood swamp in Baton Rouge, Louisiana were studied in laboratory batch systems. The effect of the age of soil contamination was studied using a laboratory-spiked soil incubated for 3 days, 3 months and 5 months. An empirical non-linear model was used to describe the bi-phasic nature of desorption with one fraction (labile) being released in relatively short periods of time (typically 24-100 hr) and a second fraction (non-labile or irreversible) being resistant to desorption. The non-linear model parameters, viz., the fraction of the chemical released rapidly (F), and the first order desorption rate coefficients, k1 and k2 respectively for the labile and slowly released fractions were determined by fitting the experimental data to the model. The data fit the model well as indicated by the high r2 values. The estimate of k1 was good. However, the values of k2 are known with less precision due to the limited duration of the experiment and number of samples taken at long times. In addition, desorption kinetics of 3 and 5-month old contaminated soils showed that progressively less amount of contaminant was available for facile desorption (lower F) compared to freshly contaminated soil. The labile fraction had desorption rate constants of the order of 10(-1) h(-1), whereas the slowly released fraction had rate constants of the order of 10(-4) h(-1) in accord with literature reported values for a variety of other compounds and soils. Possible mechanisms describing these rates and implications for the site clean up are discussed. PMID- 15900668 TI - Implants in partially edentulous patients. A longitudinal study of bridges supported by both implants and natural teeth. AB - The aim of the study was to compare the outcome of bridges supported by implants with bridges supported by a combination of implants and natural teeth abutments. The study comprised 23 patients with Applegate Kennedy Class I dentition in the mandible and a full upper denture. Implants ad modum Branemark were inserted in the posterior areas of both mandibular quadrants. On one side, a bridge supported by 2 implants was constructed (Type I) and on the other side, a bridge supported by 1 tooth (mostly the canine or first premolar) and 1 implant was made (Type II). A total of 46 bridges were made and during the 3-year follow-up period, 4 Type I and 2 Type II bridges were lost. 8 out of 69 implants were lost during the 3-year follow-up, resulting in an implant survival rate of 88.4%. Marginal bone loss, one of several parameters, was evaluated on standardized intraoral radiographs. This was performed during the 1st and 2nd year of function and the total mean bone loss from loading was 0.46 mm and 0.56 mm, respectively. The bone loss during the 2nd year of function was significantly less, adjacent to implants supporting Type II bridges, than adjacent to implants supporting Type I bridges. Summarily, no disadvantages of combining of teeth and implants in the same bridge were found in this study. On the contrary, the slightly lower marginal bone loss adjacent to implants in Type II bridges may indicate that the bone reactions could be more favorable when bridges are connected to both implants and teeth. PMID- 15900669 TI - Speech before and after treatment with bridges on osseointegrated implants in the edentulous upper jaw. AB - Speech may be affected by the transition from complete dentures to fixed prosthesis on osseointegrated implants (OIB). If so, is there any correlation between interdental spaces (open or closed) in the prosthodontic construction and deteriorated speech? 21 individuals advocated for maxillary OIB were randomised so that one half of the group got constructions with wide interdental spaces and the other half with minimised interdental spaces. All patients were examined audiologically and their speech was individually tape-recorded before and after treatment. Perceptual, acoustic, model and audiological analyses were made. Approximately 60% of the patients were judged to have indistinct speech after the treatment. The s-sound was perceptually and acoustically distorted. 67% of the patients suffered from hearing defects. No correlation between interdental spaces (open or closed) and deteriorated speech could be found. It is supposed, that hearing plays a part in the effort to adjust speech to a new maxillary OIB, and thus reduced hearing can enhance speech difficulties. PMID- 15900670 TI - Posterior jaw bone resorption in osseointegrated implant-supported overdentures. AB - For several years, osseointegrated implant-supported overdentures have been used in the rehabilitation of full edentulism with excellent results, at least in the lower jaw. This study involved 3 groups of patients with different prosthetic reconstructions: (1) mandibular overdentures supported by 2 implants connected by a bar (30 patients), (2) mandibular fixed prostheses supported by 4-6 implants (25 patients) and (3) mandibular complete dentures without implant support as controls (85 patients). The primary aim of this study was to examine on orthopantomograms (by means of the area index to minimize distortion and magnification errors), posterior mandibular ridge resorption in the 3 treatment groups. The present data demonstrated a minimal posterior mandibular ridge resorption in patients with fixed implant-supported prostheses. A more considerable posterior ridge resorption was observed in the complete denture group and especially in the overdenture wearers. For the latter, the annual posterior jaw bone resorption after the post-extraction remodeling period of 6 months, was 2- to 3-fold that of full denture wearers. When patients were edentulous for more than 10 years, the difference between the 2 latter groups disappeared. It is suggested that although the overdenture design on 2 implants offers advantages from a financial and failure rate point of view, its indications in younger patients should cautiously be evaluated in a long-term perspective concerning posterior mandibular bone resorption. PMID- 15900671 TI - Histomorphometric analyses of hydroxyapatite-coated and uncoated titanium implants. The importance of the implant design. AB - Hydroxyapatite-coated and uncoated titanium screws were inserted in the rabbit tibial metaphysis and evaluated by histomorphometry after 6 months of follow-up. There was no difference in direct bone contact between the HA-coated implants and the uncoated controls. The effect of HA-coating on screw-shaped implants seems to be uncertain, in contrast to unthreaded cylindrical designs, where there is more abundant documentation in favour of HA-coated implants. PMID- 15900672 TI - Histomorphometric and removal torque analysis for TiO2-blasted titanium implants. An experimental study on dogs. AB - The aim of the present study was to compare the anchorage of TiO2-blasted screw and cylindrical implants with conventionally used machine-produced screw and cylindrical implants inserted immediately in extraction sockets on dogs. 6 adult mongrel dogs had 3rd and 4th mandibular premolars extracted bilaterally and 24 commercial pure titanium implants were placed immediately in extraction sockets and covered with mucoperiosteum. Each dog had inserted 4 implants: 1 screw implant and 1 cylindrical implant blasted with titanium-dioxide-particles; 1 screw implant and 1 cylindrical implant with machine-produced (m.p.) surface (controls). After a healing period of 12 weeks, 16 implants from 4 animals were used for removal torque test, which demonstrated that significantly higher removal torque force was needed to unscrew the implants blasted with titanium dioxide-particles, than the normal m.p. implants. The medians for the TiO2 blasted screw and cylindrical implants were > 150 Ncm and 105 Ncm, respectively, while the values for the m.p. implants were 60 Ncm and 35 Ncm, respectively. The SEM investigation demonstrated a high irregularity of the TiO2-blasted surface compared to the machined surface. The Ra and Rz values for surface roughness were higher for the TiO2-blasted implants than for the normal m.p. implants. Histomorphometrically, the arithmetic mean of the direct bone-implant contact fraction was 69%. There was no significant difference in direct bone-implant contact length fraction between TiO2-blasted implants and the control implants. The implants blasted with titanium-dioxide-particles in this study showed a better anchorage than implants with a machine-produced surface. The screw implants showed a better anchorage than the cylindrical implants. PMID- 15900673 TI - Plasminogen activator in human gingival tissue adjacent to dental implants. AB - The installment of endosseous dental implants has become an accepted treatment procedure, and the long-term clinical results appear excellent. The composition of the soft tissue environment, however, is different from that around natural teeth. One characteristic of original junctional epithelium is its association with plasminogen activator (PA) activity. In 11 patients with a total of 30 ITI hollow-screw titanium dental implants, 16 biopsies were taken. Histologic cryostat sections were assayed for the presence of PA in the junctional epithelium. The results demonstrated that junctional epithelium around titanium implants yields PA activity in a manner very similar to that of natural teeth. The ability to produce this enzyme activity is not related to the developmental origin of the junctional cells, but to their position and function at the base of the gingival sulcus. PMID- 15900674 TI - Visualisation of the mandibular canal by different radiographic techniques. AB - 6 mandibles were radiographically examined bilaterally to visualise the mandibular canal. 5 imaging techniques were used: periapical radiography, panoramic radiography, hypocycloidal tomography, spiral tomography and computed tomography (CT). Panoramic radiographs were obtained with 2 different X-ray machines. The CT-examinations comprised direct images and standard reconstructions based on axial slices. The specimens were subsequently sectioned for contact radiography. The visibility of the mandibular canal was estimated by 3 observers at special reference points on all radiographs and classified as clearly visible, questionable visibility or not visible. The contact radiographs served as the "gold standard". The inter-observer and the intra-observer agreement were assessed by calculating the overall agreement and the x value. Direct coronal computed tomography, as well as spiral and hypocycloidal tomography, gave better visualisation of the mandibular canal than periapical and panoramic radiography. PMID- 15900675 TI - [The use of in-situ hybridization for the detection of Brachyspira spp. in pigs]. AB - Diagnosis of Brachyspira infections in swine and the differentiation of the involved bacteria is time-consuming and in most cases unsatisfactory. Detecting Brachyspira directly in the damaged Brachyspira of the large intestine could provide a direct correlation between histological lesionsa and bacterial growth. In this study we investigated whether in-situ hybridization (ISH) with a digoxigenin-labeled RNA-probe is a suitable method for detecting Brachyspira in the mucosa of the large intestine. Formalin-fixed and paraffin-embedded tissue sections of the large intestine from 78 pigs, which showed macroscopic and histological findings of Brachyspira-associated colitis, were stained with hematoxylin and eosin and Warthin-Starry silver impregnation and subjected to ISH. We used a RNA-probe with a length of 334bp, complementary to a part of the 23S rRNA of all members of the genus Brachyspira. All sections were treated with this anti-sense probe and with a sense control probe. 64 samples (82%) showed clearly positive ISH signals. Thus ISH is a suitable method for detecting Brachyspira directly within the lesions of the large intestine. The quantity of Brachyspira identified by ISH was always lower than by Warthin-Starry staining. Whether this reflects lower sensitivity of the ISH technique, or the fact that other bacteria with morphological similarities to Brachyspira were also stained by Warthin-Starry is unknown as yet. The present investigations provide a basis of further research developing specific probes to distinguish between pathogenic and non pathogenic Brachyspira species and probes detecting other bacteria with morphological similarity to Brachyspira. PMID- 15900676 TI - [Analysis of bulk milk samples using polymerase chain reaction: an additional tool for bovine viral diarrhea monitoring]. AB - Programmes for the eradication and control of infections with bovine viral diarrhea virus (BVDV) concentrate on the identification and elimination of persistently infected (PI) animals. The identification of these animals is mainly based on the detection of viral antigen using ELISA techniques. Protocols detecting viral nucleic acid using RT-PCR have been described recently. Due to high costs the German model recommends screening of animals of 9 up to 36 months of age. Screening of bulk milk samples using RT-PCR technology would allow a system independent of age. The aim of the present study was to test whether bulk milk samples (1433 including max. 50 animals each) collected in four counties of Lower Saxony are suitable for a complementary identification of PI animals via RT PCR. Thirty-one bulk milk samples derived from 27 dairy herds were BVDV positive, corresponding to 2.3 % of the herds analysed in this study. Two samples first scored doubtful. Follow up tests revealed lactating PI animals in most cases (18). In other cases the epidemiological status of the herd, i.e. high sero prevalence and/or presence of PI animals among non-lactating cattle, suggested a transient infection detected in the first bulk milk sample. These results demonstrate that monitoring of lactating cattle of any age using RT-PCR is a very sensitive, economically effective additional method for the identification of PI animals. PMID- 15900677 TI - [Factors influencing conception rate after synchronization of ovulation and timed artificial insemination--a review]. AB - This review describes factors that affect conception rate after synchronization of ovulation and timed artificial insemination. Intervals of 7 days between GnRH and PGF2alpha, 48 hours to the second GnRH treatment and a further 16 to 20 hours to the timed insemination have been proven to be most effective. Conception rates (CR) increase as lactation progresses up to 100 days in milk. Primiparous cows have higher CR than older cows. Anovular cows at the start of the synchronization protocols have poor CR. These are highest for cows started in early dioestrus. While poor body condition and some post partum and post insemination health disorders have negative effects on the CR, a significant effect of postpartum chronic endometritis could not be demonstrated. High milk yield was also not shown to have a negative effect on CR in almost all studies, while the negative effect of heat stress on fertility is also found in Ovsynch cows. However, the negative effects of high milk yields and heat stress on AI submission rates are overcome by the timed insemination protocol. PMID- 15900678 TI - Elastic properties of hoof horn on different positions in the bovine claw. AB - Hind claws of 15 adult, sound Fleckvieh cows were used for material analysis. The elastic modulus was tested in tension tests according to EN ISO 527 and ASTM D 638-03 at a universal material testing machine. Samples were taken from different segments of the bovine claw to find the differences in material properties. Samples orientation was parallel to the horn tubules and transversal, respectively. Dry matter of the test samples was determined at the time of testing. Elastic modulus values were highest with mean = 659.7 N/mm2 at the dorsal wall. Values dropped axial to 416.3 N/mm2, abaxial to 343.9 N/mm2 for longitudinal (parallel) samples and to 433.1 N/mm2 for transversal samples. The elastic modulus of the sole segment was found to be 172.1 N/mm2. No difference was calculated neither between right and left feet, lateral and medial claws, nor between longitudinal and transversal samples. PMID- 15900679 TI - [Evidence based veterinary medicine]. AB - The characteristics and advantages of evidence based (human-) medicine (EBM) are introduced. By summarising information and analysing the results of different clinical trials relating to a specific topic by expert commissions concise and advanced conclusions can be formulated. That kind of evidence (certainty that results are true) increases the explanatory power of a single trial by far. Precondition for the development of an evidence based veterinary medicine (EBVM) is an improvement of the quality, design and implementation of clinical trials. Continuous publication of these conclusions (EBVM) can support the practitioner or clinician in the decision making process for an optimal treatment. Furthermore the implementation of state of the art intervention strategy is assured. PMID- 15900680 TI - [Frequency of congenital anomalies in cattle: results from the practice in comparison with literature]. AB - The study gives a short survey of the literature of frequently observed congenital anomalies in cattle and refers to data on cases of congenital anomalies registered in a veterinary practice over a period of 8 years and in an AI (artificial insemination) station of the Rinder-Union West (RUW) during 4 years. The frequency of congenital anomalies was estimated at 0.013% in the area of the RUW. In the veterinary practice a frequency of 0.51% was found. The most prevalent anomalies were seen in legs (39% in the veterinary practice and 21% in RUW) followed by congenital anomalies of the spine (9% in the veterinary practice and 17% in RUW). Arthrogryposes were most frequent among the anomalies of the legs with 65% (veterinary practice) and 39% (RUW), respectively, of all cases of registered leg anomalies in the respective area. Umbilical hernia and atresia of segments of the intestinum were seen in 8-30% of all registered cases. The frequency of congenital anomalies differed not significantly among paternal half sib groups. A questionnaire was proposed for the registration of congenital anomalies in progeny tests of AI bulls. A series of photographs showing the most prevalent congenital anomalies is supporting the registration form. PMID- 15900681 TI - Detection of ostreid herpesvirus-1 (OsHV-1) by PCR using a rapid and simple method of DNA extraction from oyster larvae. AB - A DNA extraction procedure was developed for the detection of ostreid herpesvirus 1 (OsHV-1) using the polymerase chain reaction (PCR) in oyster larvae. The DNA extraction procedure developed was tested on 8 larval samples. Abnormal nuclei with characteristic features associated with OsHV-1 infections were only observed in samples in which the viral DNA was detected by PCR. A previously described competitive PCR method was applied to detect inhibition during PCR reactions. The results show that the method can be used on small amounts of oyster larvae (3 mg) for the detection of OsHV-1 DNA by PCR. PMID- 15900682 TI - Differences in susceptibility of palaemonid shrimp species to yellow head virus (YHV) infection. AB - Five species of palaemonid shrimp, Macrobrachium rosenbergii, M. lanchesteri, M. sintangense, Palaemon styliferus and P. serrifer, were collected from Penaeus monodon farming areas in Thailand. Some of each species were artificially infected with yellow head virus (YHV) by injection and then monitored by RT-PCR and by immunohistochemistry using monoclonal antibodies specific to 116 kDa, 64 kDa, and 20 kDa proteins of YHV. Natural YHV infections were not detected in any of the shrimp examined. In YHV injection experiments, a high proportion of P. serrifer, P. styliferus and M. sintangense exhibited mild to moderate YHV infections at 3 d post-injection. The severity of infection was reduced in shrimp that survived to 10 and 30 d post-injection. Using immunohistochemistry and RT PCR, a small proportion of M. lanchesteri showed very mild YHV infections at Day 3 but no infections at Days 10 and 30. No YHV infections resulted in M. rosenbergii. The evidence suggested that M. sintangense, P. styliferus and P. serrifer are susceptible to YHV and carry it for some time. In contrast, M, rosenbergii and M. lanchesteri appear to resist YHV infection and eliminate YHV efficiently. Because they display a range of responses to YHV, palaemonid shrimp may serve as a good model for studying YHV defense mechanisms in shrimp. PMID- 15900683 TI - Efficacy of an infectious hematopoietic necrosis (IHN) virus DNA vaccine in Chinook Oncorhynchus tshawytscha and sockeye O. nerka salmon. AB - The level of protective immunity was determined for Chinook Oncorhynchus tshawytscha and sockeye/kokanee salmon (anadromous and landlocked) O. nerka following intramuscular vaccination with a DNA vaccine against the aquatic rhabdovirus, infectious hematopoietic necrosis virus (IHNV). A DNA vaccine containing the glycoprotein gene of IHNV protected Chinook and sockeye/kokanee salmon against waterborne or injection challenge with IHNV, and relative percent survival (RPS) values of 23 to 86% were obtained under a variety of lethal challenge conditions. Although this is significant protection, it is less than RPS values obtained in previous studies with rainbow trout (O. mykiss). In addition to the variability in the severity of the challenge and inherent host susceptibility differences, it appears that use of a cross-genogroup challenge virus strain may lead to reduced efficacy of the DNA vaccine. Neutralizing antibody titers were detected in both Chinook and sockeye that had been vaccinated with 1.0 and 0.1 pg doses of the DNA vaccine, and vaccinated fish responded to viral challenges with higher antibody titers than mock-vaccinated control fish. PMID- 15900684 TI - Early interactions of Flavobacterium psychrophilum with macrophages of rainbow trout Oncorhynchus mykiss. AB - The early interactions of a low and a highly virulent Flavobacterium psychrophilum strain with head kidney and spleen macrophages of rainbow trout Oncorhynchus mykiss were characterized. The highly virulent strain was killed 5.8 to 11 times less frequently than the low virulent strain. The head kidney macrophages showed a microbicidal activity approximately twice as high as that of the spleen macrophages. A 2- to 3-fold higher production of reactive oxygen species (ROS) was induced by the highly virulent strain than by the low virulent one. The head kidney macrophages produced approximately twice as much ROS as the spleen macrophages. The low virulent strain was killed approximately 10 times more frequently by H2O2 than was the highly virulent strain. In spleen macrophages, the highly virulent strain caused twice as much cytotoxic effects compared to the low virulent strain. In conclusion, virulence in F. psychrophilum appears to be correlated with higher O. mykiss macrophage cytotoxicity and resistance to ROS and, therefore, with enhanced resistance to bacterial killing. Moreover, due to lower ROS production, spleen macrophages have a lower antimicrobial action against F. psychrophilum, compared to head kidney macrophages and, thus, might form a 'safe site' in which bacteria can reside. PMID- 15900685 TI - Detection of mycobacteria in aquarium fish in Slovenia by culture and molecular methods. AB - Thirty-five aquarium fish were investigated for the presence of mycobacteria by culture and molecular methods. The following species were examined: goldfish Carassius auratus auratus, guppy Poecilia reticulata, 4 three-spot gourami Trichogaster trichopterus, dwarf gourami Colisa lalia, Siamese fighting fish Betta splendens, freshwater angelfish Pterophyllum scalare, African cichlid fish Cichlidae spp., cichlid fish Microgeophagus altispinosus, cichlid fish Pseudotropheus lombardoi, blue streak hap Labidochromis caeruleus, sterlet Acipenser ruthenus, southern platyfish Xiphophorus maculatus, and catfish Corydoras spp. Isolates of mycobacteria were obtained in 29 cases (82.9%). Two specimens were positive using Ziehl-Neelsen (ZN) staining, but the cultivation failed. Four specimens were both ZN- and culture-negative. On the basis of GenoType Mycobacterium assay (Hain Life-science) and restriction enzyme analysis of the amplified products (PCR-RFLP), 23 isolates (79.3%) were identified: 7 as Mycobacterium fortuitum, 6 as M. gordonae, 6 as M. marinum, 3 as M. chelonae, and 1 as M. peregrinum. Five isolates remained unidentified (Mycobacterium spp.). One case probably represented a mixed infection (M. marinum/M. fortuitum). Since M. marinum infections are also detected in humans, the significance of mycobacteria in aquarium fish should not be overlooked. PMID- 15900686 TI - Characterisation of an emerging rickettsia-like organism in Tasmanian farmed Atlantic salmon Salmo salar. AB - A rickettsia-like organism (RLO) was observed in farmed Atlantic salmon Salmo salar located in south-east Tasmania, Australia. Several assays such as immunoperoxidase, immunoelectron microscopy, polymerase chain reaction and nucleic acid sequencing, as well as phylogenetic analysis of rDNA sequences, were performed on infected fish tissues. Immunohistochemistry results suggested the presence of related antigenic determinants between the Tasmanian RLO and the type strain LF-89 of Piscirickettsia salmonis. However, sequence alignment demonstrated that the Tasmanian RLO contains a 19 bp deletion at the 3'-end of the internal transcribed spacer region of the rDNA operon, indicating a genetic divergence from P. salmonis isolates, which are exotic to Australia. PMID- 15900687 TI - Histochemistry of glycoconjugates in mucous cells of Salmo trutta uninfected and naturally parasitized with intestinal helminths. AB - Mucus secreted onto the surface of the intestine forms a physical barrier to invading parasites so that a possible attachment of helminths to the surface is prevented and their expulsion by peristalsis facilitated. In mammals, intestinal parasites induce hyperplasia and hypertrophy of intestine goblet cells and provoke changes in the mucus composition. In fish, this topic has received less attention. In the present investigation, histochemical methods were employed to compose intestinal mucous cell numbers and their glycoconjugate composition were compared by uninfected brown trout Salmo trutta and in S. trutta parasitized with Cyathocephalus truncatus or Pomphorhynchus laevis. When P. laevis was present in the intestine of the brown trout, the total mucous cell number, and the number of mucous cells containing acid or mixed glycoconjugates were significantly enhanced. No significant change in the total mucous cell number was detected in the intestine of fish parasitized with C. truncatus in comparison with uninfected brown trout. A significant increase was observed in the number of both acid (especially sulphated) and mixed glycoconjugates containing mucous cells as well as a significant decrease in the number of neutral glycoconjugates containing mucous cells. When intestinal helminths were present, the thickness of the adherent mucous gel increased. In a limited number of other fish species, the occurrence of gill and intestinal parasites has been reported to increase the mucosal glycoconjugate secretions. Our study is the first quantitative report on the effects of intestinal helminths on the density of mucous cells and mucus composition in a fish species. PMID- 15900688 TI - Radiodiagnostic method for studying the dynamics of Anguillicola crassus (Nematoda: Dracunculoidea) infection and pathological status of the swimbladder in Lake Balaton eels. AB - Swimbladder changes resulting from Anguillicola crassus infection of the European eel Anguilla anguilla have been the subject of several studies reported in the literature. These investigations, however, studied exclusively the status of infection at a given point in time and did not deal with changes in swimbladder infection in eels suffering from anguillicolosis over a period of time. In this study, A. crassus-induced pathological changes were monitored in 78 eels naturally infected in Lake Balaton and subsequently kept in the laboratory, thus excluding the possibility of further infection. During the 3 mo study, the status of the swimbladder was checked by radiographic examination on 4 occasions. At the end of the study the eels were dissected and the gross pathological changes in the swimbladders were compared with the radiographic findings. As compared to their starting condition, by the end of the study the pathological status of the swimbladder had deteriorated in 55% and remained the same in 37% of the cases. Tendency to improvement (1%) and variable findings (7%) were recorded in a low percentage of cases only. With the help of the radiographs presented, the dynamics of A. crassus infection and of changes in the swimbladder of individual eel specimens can be monitored easily. PMID- 15900689 TI - Classification and occurrence of abnormally developed Paradiplozoon homoion (Monogenea, Diplozoinae) parasitising gudgeon Gobio gobio. AB - Morphological analyses of the attachment apparatus (clamps and central hooks) of Paradiplozoon homoion (Bychowsky & Nagibina, 1959) (Diplozoinae, Monogenea) parasitising gills of Gobio gobio (L.) showed a high percentage of abnormally developed parasite specimens. Four different localities in the Vlara River basin, Czech Republic, were investigated for the presence of such abnormal individuals. The highest percentage of abnormalities in the attachment apparatus (over 39%) was recorded in the Vlara River, at Bohuslavice. This study provides a comprehensive classification of these abnormalities with 7 types of abnormalities described and illustrated. Abnormalities of parts of the attachment apparatus that form in the later stage of ontogenetic development were the most frequent, the most frequent types of abnormalities being clamps with abnormal sclerites, and combinations of abnormalities. Abnormalities of the central hooks were also found in our material. The abnormalities found in diplozoids are probably connected with environmental pollution; however, this point requires further investigation. PMID- 15900690 TI - Treatment of ichthyophthiriasis after malachite green. I. Concrete tanks at salmonid farms. AB - Since the use of malachite green was banned in many European countries, new alternative treatments have been tested to prevent white spot disease caused by Ichthyophthirius multifiliis. We tested formalin, potassium permanganate (KMnO4), chloramine-T, hydrogen peroxide (H2O2) and Per Aqua or Desirox alone or in combinations of 2 chemicals, one of which was always formalin, in 50 m2 concrete tanks at 2 farms producing salmon Salmo salar smolt in 2001 and 2002. Both Per Aqua and Desirox are combinations of peracetic acid, acetic acid and hydrogen peroxide. The alternative chemicals or their combinations can be used successfully to lower the parasite burden to such a level that no high mortality occurs during the first 4 wk after the start of an infection. This period of time allows the fish to develop immunity against these ciliates, and treatments can be reduced and stopped in due course. I. multifiliis decreased in number 3 to 4 wk after the beginning of the infection in all the treatments. Large differences in parasite burden and mortality occurred among the replicates in all except the Desirox-formalin tanks, which means that they are not as reliable as the malachite green-formalin used previously. It was also evident that the chemicals and their concentrations must be planned carefully to suit the conditions on each farm. PMID- 15900691 TI - Parasite communities of Adriatic cage-reared fish. AB - From June 2001 to March 2002, 7 semi-offshore facilities in the Adriatic Sea rearing sea bass Dicentrarchus labrax, sea bream Sparus aurata, sharpsnout bream Diplodus puntazzo and red sea bream Pagellus bogaraveo aged 1+, were monitored for the presence of protozoan and metazoan parasites. Obtained data sets of abundance and prevalence were used to evaluate the structure and dynamic of infra and component parasite communities. In all hosts, except the red sea bream, dominant parasites were monogenean specialists, showing clear seasonality. Average infracommunity richness was very low, ranging from 1 to 3 parasites per fish, while at component community level, values ranged from 3 to 8, depending on host species. Based on low diversity indices and a moderate to high similarity coefficient between different facilities, it can be concluded that the parasitofauna of Adriatic cage-reared fish is a stable and impoverished stochastic assemblage, with monogeneans as the predominant parasitic group. PMID- 15900692 TI - Occurrence of Perkinsus olseni (Protozoa: Apicomplexa) and other parasites in the venerid commercial clam Pitar rostrata from Uruguay, southwestern Atlantic coast. AB - A study was conducted into the health status of natural populations of the venerid clam Pitar rostrata from Uruguay. Perkinsus sp. was detected in 22% of the clams. Severe hemocytic infiltration was detected in the tissues parasitized by this protozoan parasite. The sequencing of the ITS-5.8S gene cluster of the parasite confirmed that it belonged to the Perkinsus olseni species. Rickettsia or Chlamidia-like organisms were also found, with a prevalence of 11%, although without apparent host reaction; an unidentified species of Coccidia was found in the nephridia of 78% of the clams, with the intensity of infection ranging from moderate to high. A gregarine, Nematopsis-like organism was observed mainly in the epithelial cells of the intestine, without host response and with a prevalence of 56%. Of the metazoan parasites, trematodes were found in 11% of the individuals analyzed. PMID- 15900693 TI - Relationship between papillary fibrosis and patency of the accessory pancreatic duct. AB - A study of structure and function of the minor duodenal papilla was carried out on 36 autopsy specimens of the human duodenopancreas. An original perfusion method with manometric control was developed for testing the minor papilla patency, i.e. resistency to intraductal pressure. According to this test, all the specimens were classified as: Type I: non-patent (45%); Type II: patent under normal, physiological pressure (36%); and Type III: patent under increased pressure (19%). After that, the whole minor papilla specimens underwent longitudinal sectioning with HE staining. Light microscopy showed various degrees of fibrosis (mild-45%, moderate-29%, and severe-26%). The correlation between the fibrosis and the patency was significant: papillae with lower degrees of fibrosis were more patent and vice versa. Further, the pancreatic tissue in the Santorini segment of the pancreatic head followed the same manner. No apparent sphincter of the minor papilla was found in any of the specimens analyzed. PMID- 15900694 TI - The anatomical study on the rat pancreas and its ducts with emphasis on the surgical approach. AB - It was intended to present information about the anatomy of the pancreas and especially to emphasize the variation of pancreatic ducts in the rat, which may guide researchers in experimental studies. In 27 adult rats, latex dye was introduced into the biliopancreatic duct, portal vein and arteries. The pancreas with the neighboring structures was studied with the aid of the dissection microscope and measurements were by means of micrometric ocular. The pancreas was divided into three parts i.e. the biliary, duodenal and gastrosplenic portions. The biliopancreatic and pancreatic ducts as well as biliary and duodenal portions of the pancreas could be seen ventrally with a minor procedure such as pulling the duodenum caudally, and additionally the entire pancreas and its ducts could also be reached dorsally since the stomach was turned cranially with the duodenum. The biliopancreatic duct diameter and length of the ducts were 1.01+/ 0.03 and 28.86+/-0.59 mm, respectively. The anterior pancreatic duct originated from the biliopancreatic duct on different sides, 17.96+/-0.75mm away from the duodenal papilla. The anterior pancreatic duct drained the gastrosplenic portion of the rat pancreas and this duct, via the duodenal duct, partially collected secretion of the duodenal portion at a ratio of 74.07%. The posterior pancreatic duct opened to the biliopancreatic duct and its distance from papilla duodeni was changed from 0 to 8 mm. When the biliopancreatic duct is ligated at the level of the duodenal opening, the posterior pancreatic duct or a small duct from duodenal portion should be taken into consideration because these ducts opened to biliopancreatic duct at the level of papilla duodeni at a ratio of 37.04%. In conclusion, the duct system of pancreas had a great variation and therefore the success rate of the surgery could be increased when surgeons have more knowledge on variations of the duct system in this region. PMID- 15900695 TI - Long-term reinnervation effects after sciatic nerve lesions in adult rats. AB - Transection of the sciatic nerve in adult rats induces drastic changes in hindleg muscles. Earlier, we demonstrated that the reinnervated soleus (SOL) muscle, 21 weeks after a transection mainly contains type II fibers. This is in striking contrast to normal muscle, which consists to 80% of type I muscle fibers. Also we observed 13.9% of the fibers to be polyneurally innervated. The problem of the present study is whether these changes are reversible after Longer survival periods. Therefore, the SOL was studied 60 weeks after transection and reconstruction by an autologous nerve graft. In six rats, we studied muscle fiber distributions by monoclonal antibodies, and innervation patterns by cholinesterase staining and AgNO3 impregnation. Still at 60 weeks, only 20% of the muscle fibers are of type I and this is similar to results at 21 weeks, indicating that no recovery to the normal has been reached by that age. Furthermore, 20% of the endplates in the reinnervated SOL were polyneurally innervated, but we also observed this in 10% of the endplates on the control side. These increases, compared to data at 21 weeks, are interpreted as an aging effect. PMID- 15900696 TI - Bilateral absence of ansa cervicalis replaced by vagocervical plexus: case report and literature review. AB - The bilateral absence of the ansa cervicalis was discovered during dissection of the neck in an elderly male cadaver. On both sides it was replaced by a vagocervical plexus formed by the vagus nerve and C1 and C2 ventral rami from the cervical plexus. A descending branch from this vagocervical plexus supplied the strap muscles of the neck. From a review of the literature the incidence of bilateral absence of the ansa cervicalis and its replacement by a vagocervical plexus appears to be extremely rare. Morphological variability, embryological basis, and clinical relevance of this uncommon anatomical variant are discussed. PMID- 15900697 TI - [Micro anatomy of the cavernous sinus]. AB - PURPOSE: The surgical anatomy of the human cavernous sinus has been described by several authors. A clearer understanding of the anatomy of the cavernous sinus has resulted in an increased interest in surgical approaches to this region. Regarding the anatomical concept of the cavernous sinus previous descriptions should be revised. METHODS: In our study, 35 skull-base blocks including both cavernous sinuses were investigated. RESULTS: There is considerable reason that the lateral wall of the cavernous sinus is covered by a "medial temporal dura layer" attached to the leptomeningeal pockets of the cranial nerves III-V1+2. There is no significant constance of the internal carotid artery within the cavernous sinus. All cranial nerves adjacent to or traversing the cavernous sinus are embedded within a leptomeningeal sheath similar to a pocket. The abducent nerve traverses a venous space, which should be named "petroclival venous confluence" rather than Dorello's canal. The extremely variable cavernous space is covered by a one-layer cell line. There is no evidence of any venous-plexus structure. Fibrose bands (trabecle) are present in different numbers. PMID- 15900698 TI - A morphometric study and variations on the lumbar arteries of human fetuses. AB - Lumbar arteries are in series with the posterior intercostal arteries. The aim of this study has been to investigate the morphometric data on the abdominal aorta of the human fetus and define different types of origin variations of the lumbar artery. Initially, the latex solution colored with red ink was injected into the thoracic aorta. The abdominal aorta and its branches were dissected and examined bilaterally in 120 fetuses (ranging between 16 and 32 post-menstrual weeks) and the anatomic variations recorded. Fourteen fetuses with variations were studied and photographed. The origins and morphologic variations of the lumbar arteries were defined and classified. The morphological relationship of the lumbar arteries to the abdominal aorta and the length and width (diameter) of the vessels were investigated by performing measurements using a digital calliper (mm). The frequency and the types of the different variations determined in the present study have been listed. Abdominal aortic branches and Lumbar artery aneurysms are rare lesions with potentially life-threatening consequences and they are difficult to access anatomically and formidable to manage operatively. During the performance of conventional or open surgical replacement of an abdominal aortic aneurysm with prosthetic grafts, the surgeon needs precise knowledge of the anatomy of the abdominal aortic branches and immediate retroperitoneal structures. The variations on the lumbar arteries may have clinical importance. When this region is under diagnostic and/or surgical investigation using computed tomography scan or diagnostic angiography, the possibility of these variations should be take into consideration to avoid complications. PMID- 15900699 TI - Skeletal segments of the human pollical and hallucal rays: comparison and analysis of their intrinsic proportions. AB - Since in humans, the first digital ray has a different functional significance in the hand and in the foot, the comparison of the pollical and hallucal rays seems interesting in order to evaluate the influence of specific evolutionary functional factors and biomechanical constraints. The objective of this study was to provide original and detailed data on the intrinsic proportions of the three segments of the human pollical and hallucal rays (metacarpal or metatarsal bone, proximal phalanx, and distal phalanx) in order to allow a quantitative comparison of their relative development. No similar data evaluated from three-dimensional approaches (volumetric or ponderal) seem to be available in the literature. The material consisted of 77 skeletons of the pollical ray and 77 skeletons of the hallucal ray of normal adult individuals. The ponderal approach was used. The first metapodial represented a mean of 57.17% for the pollical ray and 68.48% for the hallucal ray, whereas the phalanges respectively represented 42.83% and 31.52%. The intrinsic phalangeal proportions were almost identical for the pollex and for the hallux (respectively, 71.76% and 72.88% for the proximal phalanx; and 28.24% and 27.12% for the distal phalanx). Since these intrinsic proportions are the reflection of biological, functional and evolutionary constraints, these data seem of interest for the analysis of the functional significance of these rays, for the characterization of malformative syndromes in clinical practice, for reference in comparative morphology, and for the interpretation of hominid fossil bones. PMID- 15900700 TI - Accessory muscle in the hypothenar region: a functional approach. AB - An accessory muscle was found in the hypothenar region on both hands during routine cadaver dissection. This muscle originated from the tendon of the flexor carpi radialis, crossed the palma manus region superficially and inserted together with the abductor digiti minimi muscle into the ulnar aspect of the basis of the fifth proximal phalanx. The muscle was supplied by one branch arising from the main trunk of the ulnar nerve. Abnormalities of the hypothenar muscles have been described by many authors with a focus on their structural aspects, but there is not enough data about the possible functions they could induce. In our study, we try to elucidate the functions of this accessory muscle. We did not name the variant muscle as it has various functions, each similar to that of individual hypothenar muscles. PMID- 15900701 TI - Adaptation of the hindlimbs for climbing in bears. AB - The hindlimbs of the Malayan sun bear (Helarctos malayanus), the polar bear (Ursus maritimus), the brown bear (Ursus arctos) and the giant panda (Ailuropoda melanoleuca) have been anatomically and osteometrically studied. The Musculus tibialis cranialis of the Malayan sun bear and the giant panda possessed a well developed rich fleshy portion until the distal end of the tibia. In the polar bear and the brown bear, however, the fleshy portion of the M. tibialis cranialis was not developed until the distal end of the tibia. The tendon of the M. tibialis cranialis inserting on the proximal end of the Ossa metatarsalia was shorter in the Malayan sun bear and the giant panda than in the polar bear and the brown bear. In the Malayan sun bear and the giant panda, moreover, the M. popliteus was attached more distally to the tibia than in the polar bear and the brown bear. The stable dorsiflexion and supination of the foot and the efficient pronation of the crus are important for skillful tree climbing. The present study suggests that the Malayan sun bear and the giant panda have hindlimbs especially adapted to tree climbing by the well-developed fleshy portion of the M. tibialis cranialis reaching the distal end of the tibia, its short tendon, and the M. popliteus inserting near the distal end of the tibia. PMID- 15900702 TI - Postnatal cerebellar granule cells of the white rat (Rattus norvegicus): a quantitative study, using design-based stereology. AB - A stereological study was carried out on postnatal cerebellar granule cells of rats aged 6 and 10 days, for detecting whether and how much they would differ from those of young adult rats. The following parameters were estimated: number weighted mean volume of the nucleus and of the soma; mean total surface area of the soma; mean absolute volumes per cell of total cytoplasm, mitochondria, Golgi apparatus, and cytosol; mean surface density of the rough endoplasmic reticulum (RER); mean total surface area of the RER. These values were compared between the two postnatal ages. In addition, those values were also analysed in comparison to the ones depicted in young adult rats (60 days), already published by our team, in order to detect similarities between them. It was noticed that, between 6 and 10 days, the mean surface density of the RER was the only parameter that did not change significantly. The comparison of each of the postnatal ages with 60 days revealed that, with the exception of the absolute volume of Golgi apparatus, significant differences were displayed concerning other organelles and cellular compartments. It was concluded that, although fine structural differences have been disclosed, from the stereological point of view postnatal granule cells at 10 days were practically similar to the young adult ones at 60 days. Some potential physiological implications have been considered. PMID- 15900703 TI - Lymphocyte-rich Hassall bodies in the normal human thymus. AB - The study is concerned with the presence of an unusual Lymphocyte-rich type of Hassall body in the normal human thymus. Lymphocyte-rich Hassall bodies are large, they are usually lined by squamous epithelial cells, and contain closely packed lymphocytes in areas with necrotic and hyaline material. Lymphocyte-rich areas of the HassaLL bodies do not contain epithelial cells (as demonstrated with cytokeratin 8/18, 7, 34betaE12, and MNF116), they were negative for CD20 and S100 protein. Immunoreaction for CD3 was positive in 3 of 4 cases. Our results suggest that this type of Hassall body may be involved in the negative selection of thymocytes. PMID- 15900704 TI - A geometric description of the urinary bladder of Turkish shepherd dog (Karabas). AB - In this study, a geometric and experimental work of the urinary bladder of a dog is presented. Experimentally, the diameters on the neck (collum vesicae region), the body (corpus vesicae region), the bottom (apex region) and the longitudinal length of the urinary bladder were measured. Geometrically it was shown that the urinary bladder is comparable with a surface of revolution of a planar (profile) curve. On the same regions of the modelled surface the diameters of the horizontal sections have been measured. It was found that the geometric and the experimental values were closely related. Further the Gaussian curvature K at certain points on the same regions of the modelled surface was calculated. The quadratic approximations of the urinary bladder surface was described with the sign of K at those points. PMID- 15900705 TI - Light microscopic study of periapical lesions associated with asymptomatic apical periodontitis. AB - The purpose of the study has been to evaluate the degree of chronic inflammation in tissues surrounding the apex of the tooth root in patients with apical periodontitis in the remission phase. The material included 37 apical granulomas and radicular cysts obtained as a result of apiectomy, and 20 teeth which were removed together with the focus of the periapical inflammation. Routine histological techniques, as well as the immunofluorescent and immuno-chemical methods were used to examine the material. Despite the absence of clinical symptoms in 23 of 57 cases, the morphological signs of chronic inflammation were observed in the apical area of the tooth root. Morphological signs of viral invasion of epithelial and stromal cells in the radicular cyst wall were revealed in six cases. The presence of the virus of Herpes simplex I in epithelial cells (five cases) and adenoviral invasion (one case) was confirmed by immuno fluorescent and immuno-chemical methods. Histological examination often reveals morphological signs of an active inflammatory process in the periapical tissues of patients treated during clinical remission. In our opinion, the presence of viruses in the epithelial cells of the radicular cyst may contribute to the persistence of the active stage of the inflammatory process. PMID- 15900706 TI - The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles. AB - Synaptophysin and synaptobrevin are abundant membrane proteins of neuronal small synaptic vesicles. In mature, differentiated neurons they form the synaptophysin/synaptobrevin (Syp/Syb) complex. Synaptobrevin also interacts with the plasma membrane-associated proteins syntaxin and SNAP25, thereby forming the SNARE complex necessary for exocytotic membrane fusion. The two complexes are mutually exclusive. Synaptobrevin is a C-terminally membrane-anchored protein with one transmembrane domain. While its interaction with its SNARE partners is mediated exclusively by its N-terminal cytosolic region it has been unclear so far how binding to synaptophysin is accomplished. Here, we show that synaptobrevin can be cleaved in its synaptophysin-bound form by tetanus toxin and botulinum neurotoxin B, or by botulinum neurotoxin D, leaving shorter or longer C terminal peptide chains bound to synaptophysin, respectively. A recombinant, C terminally His-tagged synaptobrevin fragment bound to nickel beads specifically bound synaptophysin, syntaxin and SNAP25 from vesicular detergent extracts. After cleavage by tetanus toxin or botulinum toxin D light chain, the remaining C terminal fragment no longer interacted with syntaxin or SNAP 25. In contrast, synaptophysin was still able to bind to the residual C-terminal synaptobrevin cleavage product. In addition, the His-tagged C-terminal synaptobrevin peptide 68 116 was also able to bind synaptophysin in detergent extracts from adult brain membranes. These data suggest that synaptophysin interacts with the C-terminal transmembrane part of synaptobrevin, thereby allowing the N-terminal cytosolic chain to interact freely with the plasma membrane-associated SNARE proteins. Thus, by binding synaptobrevin, synaptophysin may positively modulate neurotransmission. PMID- 15900707 TI - Disruption of target cell adhesion structures by the Yersinia effector YopH requires interaction with the substrate domain of p130Cas. AB - The docking protein p130Cas has, together with FAK, been found as a target of the Yersinia virulence effector YopH. YopH is a protein tyrosine phosphatase that is delivered into host cells via the bacterial type III secretion machinery, and the outcome of its activity is inhibition of host cell phagocytosis. In the present study using p130Cas-/- cells, and p130Cas-/- cells expressing variants of GFPp130Cas, we show that this docking protein, via its substrate domain, is responsible for subcellular targeting of YopH in eukaryotic cells. Since YopH inhibits phagocytosis, p130Cas was expected to be critical for signalling mediating bacterial internalization. However, p130Cas-/- cells did not exhibit reduced capacity to internalize Yersinia. On the other hand, when a dominant negative variant of p130Cas was expressed in these cells, the phagocytic capacity was severely impaired. Moreover, the p130Cas-/- cells displayed a marked reduced sensitivity towards YopH-mediated detachment compared to wild-type cells. Transfecting these cells with full-length p130Cas rendered cells hypersensitive to both mechanical and Yersinia-mediated detachment. This hypersensitivity was not seen upon transfection with the dominant negative substrate domain-deleted variant of p130Cas. This implicates p130Cas as a prominent regulator of cell adhesion, where its substrate-binding domain has a significant function. PMID- 15900708 TI - Hid can induce, but is not required for autophagy in polyploid larval Drosophila tissues. AB - The major cell death pathways are apoptosis and autophagy-type cell death in Drosophila. Overexpression of proapoptotic genes in developing imaginal tissues leads to the activation of caspases and apoptosis, but most of them show no effect on the polytenic cells of the fat body during the last larval stage. Surprisingly, overexpression of Hid induces caspase-independent autophagy in the fat body, as well as in most other larval tissues tested. Hid mutation results in inhibition of salivary gland cell death, but the disintegration of the larval midgut is not affected. Electron microscopy shows that autophagy is normally induced in fat body, midgut and salivary gland cells of homozygous mutant larvae, suggesting that Hid is not required for autophagy itself. Constitutive expression of the caspase inhibitor p35 produces identical phenotypes. Our results show that the large, post-mitotic larval cells do not react or activate autophagy in response to the same strong apoptotic stimuli that trigger apoptosis in small, mitotically active imaginal disc cells. PMID- 15900709 TI - Activated cofilin colocalises with Arp2/3 complex in apoptotic blebs during programmed cell death. AB - Etoposide inhibits topoisomerase II and induces apoptosis in human epidermoid cancer cells (A431) and normal rat fibroblasts (NRK) as verified by apoptotic morphology and chromatin degradation. Here we examine changes in the localisation of actin, cofilin and the Arp2/3 complex during the apoptotic process in response to etoposide. Twenty-four hours after etoposide addition, a large number of cells of both lines exhibited nuclear and cytoplasmic fragmentation with the formation of numerous blebs typical of apoptosis. Etoposide exposure induces dissolution of stress fibres and an increase in actin and cofilin in membrane patches and apoptotic blebs. The actin is more peripherally located than the cofilin, similar to that reported for lamellipodia of highly motile keratocytes. By contrast, in control cells, cofilin is evenly distributed throughout the cytoplasm, though often enriched around the nucleus. The active form is inferred to be more peripherally localised and to be present in apoptotic blebs, since an antibody specific for phosphorylated cofilin did not stain the cell periphery nor apoptotic blebs. Although immunoblots of 2D gels demonstrate that the ratio of de phosphorylated to phosphorylated cofilin does not change after etoposide treatment, this does not mean that there are no changes in the turnover of the active and inactive forms. Transfection of both cell lines with EGFP-containing constructs of wild-type cofilin and mutants resembling its activated (S3A) and inactivated (S3D) forms shows that the active form has a more peripheral localisation and is also present in the membrane blebs with a strong colocalisation with actin. We further show that Arp2/3 also localises in apoptotic blebs and discuss the role of these proteins in apoptosis by analogy with actin-based protrusive motility in lamellipodia. PMID- 15900710 TI - Recombinant antibody expression vectors enabling double and triple immunostaining of tissue culture cells using monoclonal antibodies. AB - Next to the already available mouse monoclonal and laboratory animal-derived polyclonal antibodies, recombinant antibodies offer an additional and virtually unlimited arsenal of new immunohistochemical research tools. The major advantages of recombinant antibodies are their rapid and easy generation against virtually any target. The avidity of antibody fragments can be increased by partial dimerisation. This can be achieved by fusion of CL domains derived of different species to recombinant antibody domains. The VL-linker-VH-CL constructs result in significantly lower dimerisation levels compared to the VH-linker-VL-CL antibody constructs. The most efficient dimerisation occurs with the Jun-tagged scFvs. The very large and rapidly expanding collection of recombinant antibodies already available combined with the ease of introducing various tag sequences allows for an almost unrestricted number of easily adjustable research tools. To our best knowledge we report for the first time that using CL domains derived from different species, in combination with readily available commercial secondary antibodies specific for these CL domains, provides an easy method for the application of recombinant monoclonal antibodies of various origins in immunohistochemical analyses eliminating the problem of co-staining with multiple mono- or polyclonal antibodies. Both double and triple labelling experiments can be performed successfully. PMID- 15900711 TI - Cytoskeletal dynamics--6th Young Scientists Meeting of the German Society for Cell Biology (DGZ) held in Heidelberg, September 30th to October 2nd, 2004. PMID- 15900712 TI - Cadmium adsorption in montmorillonite as affected by glyphosate. AB - Behaviors of soil heavy metals are often affected by coexisting herbicides due to their physical and chemical interaction. Effect of glyphosate, an herbicide containing -PO3(2-) and -COOH groups, on cadmium adsorption in montmorillonite was studied in detail. The results showed that the cadmium adsorption quantity in montmorillonite increased with increasing soil solution pH and cadmium concentration as usual, but decreased with glyphosate, which is due to the formation of a low affinity complex of Cd and glyphosate and decreasing solution pH induced by glyphosate addition. When the equilibrium solution pH was below 6.7, glyphosate has little effect on cadmium adsorption, but when the equilibrium solution pH was above 6.7, glyphosate significantly decreased cadmium adsorption quantity in montmorillonite. In addition, the adding order of Cd and glyphosate also influenced Cd adsorption quantity in montmorillonite. PMID- 15900713 TI - Chemical speciation and mobility of heavy metals in municipal solid waste incinerator fly ash. AB - Chemical speciation is a significant factor that governs the toxicity and mobility of heavy metals in municipal solid waste incinerator fly ash. Sequential extraction procedure is applied to fractionate heavy metals (Pb, Zn, Cd, Cu, and Cr) into five defined groups: exchangeable, carbonate, Fe-Mn oxide, organic, and residual fractions. The mobility of heavy metals is also investigated with the aid of toxicity characteristic leaching procedure. In the fly ash sample, Pb is primarily presented in the carbonate (51%) and exchangeable (20%) fractions; Cd and Zn mainly exist as the exchangeable(83% and 49% respectively); Cu is mostly contained in the last three fractions (totally 87%); and Cr is mainly contained in the residual fraction (62%). Pb, Zn and Cd showed the high mobility in the investigation, thus might be of risk to the natural environment when municipal solid waste incinerator fly ash is landfilled or reutilized. PMID- 15900714 TI - Pyrolysis of EVA and its application in recycling of photovoltaic modules. AB - The basic pyrolysis behaviour of ethylene vinyl acetate (EVA) copolymer, which is often used as a lamination agent in solar modules, was investigated in thermogravimetry, differential thermal analysis(DTA) and thermovolumetry. The TG analysis showed that the EVA pyrolysis can be accelerated under the partial oxidizing atmosphere but the end pyrolysis temperature must be higher than in nitrogen, to eliminate the coke formed. Meanwhile, a strong exothermal peak occurs at about 450 degrees C under the air condition and gets weaker obviously at the oxygen content lower than 10 vol. %. The mass balance of EVA pyrolysis was given through the thermovolumetry with the output of 10 wt. % permanent gas, 89.9 wt. % condensate and 0.1% residual coke. Besides, the composition of the permanent gas and condensate at different pyrolysis stages were analysed and interpreted on the known pyrolysis mechanism. PMID- 15900715 TI - Screening of flocculant-producing microorganisms and flocculating activity. AB - A strain saccharomycete STSM-1 with high flocculanting activity was isolated from activated sludge with conventional methods. The high production rate and the low cost STSM-1 medium was obtained by selecting different kinds of media, carbon source, nitrogen source and inorganic salt ion. The best flocculant-producing conditions were found by changing medium initial pH, culture temperature and ventilation flow. The best flocculating effect was obtained by changing positive ion types, density and concentration of flocculant. PMID- 15900716 TI - Investigation of factors on a fungal biofilter to treat waste gas with ethyl mercaptan. AB - The biofilter is cost-effective for the waste gases treatment. The bacterial is the main microorganism in the conventional biofilters. However, it faces some problems on the elimination of hydrophobic compounds. In order to overcome these problems, the biofilters with fungi were developed. The objective of this study is to investigate the factors affecting ethyl mercaptan (EM)-degradation using a fungal biofilter. A laboratory experiment was set up. The effects of loading rate, empty bed residence times (EBRT) and pH on EM degradation were investigated. Over 95% removals of EM could be achieved, under the condition of the influent loadings below 50 g/(m x h). Removal efficiencies improved to 98% with EM loading decreased to 45 g/(m x h). For long EBRT of 58 s corresponding to a low rate of 0.3 m3/h, the EM removal efficiencies of over 98% were observed. However, when EBRT was decreased to 14 s, the removal efficiencies fell under 80%. The pH range of 3-5 was feasible to fungi. PMID- 15900717 TI - Nitrogen removal influence factors in A/O process and decision trees for nitrification/denitrification system. AB - In order to improve nitrogen removal in anoxic/oxic(A/O) process effectively for treating domestic wastewaters, the influence factors, DO(dissolved oxygen), nitrate recirculation, sludge recycle, SRT(solids residence time), influent COD/TN and HRT(hydraulic retention time) were studied. Results indicated that it was possible to increase nitrogen removal by using corresponding control strategies, such as, adjusting the DO set point according to effluent ammonia concentration; manipulating nitrate recirculation flow according to nitrate concentration at the end of anoxic zone. Based on the experiments results, a knowledge-based approach for supervision of the nitrogen removal problems was considered, and decision trees for diagnosing nitrification and denitrification problems were built and successfully applied to A/O process. PMID- 15900718 TI - Sediment distribution pattern mapped from the combination of objective analysis and geostatistics in the large shallow Taihu Lake, China. AB - Investigation was made into sediment depth at 723 irregularly scattered measurement points which cover all the regions in Taihu Lake, China. The combination of successive correction scheme and geostatistical method was used to get all the values of recent sediment thickness at the 69 x 69 grids in the whole lake. The results showed that there is the significant difference in sediment depth between the eastern area and the western region, and most of the sediments are located in the western shore-line and northern regimes but just a little in the center and eastern parts. The notable exception is the patch between the center and Xishan Island where the maximum sediment depth is more than 4.0 m. This sediment distribution pattern is more than likely related to the current circulation pattern induced by the prevailing wind-forcing in Taihu Lake. The numerical simulation of hydrodynamics can strong support the conclusion. Sediment effects on water quality was also studied and the results showed that the concentrations of TP, TN and SS in the western part are obviously larger than those in the eastern regime, which suggested that more nutrients can be released from thicker sediment areas. PMID- 15900719 TI - Spatial distribution of heavy metals of agricultural soils in Dongguan, China. AB - Distribution and speciation of heavy metals of agricultural soils(85 surface soil samples and 4 soil profiles) in Dongguan were investigated, while total Cr, Cu, Ni, Pb, Zn(abbreviated as Cr, Cu, Ni, Pb, Zn) and available Cu, Zn(Av-Cu, Av-Zn) were analyzed by a flame absorption spectrophotometer(AAS), and total Cd(Cd) was analyzed using graphite furnace AAS. The content of Cd, Cu and Ni was partially much more than the second grade of GB15618-1995 even though the mean contents of all heavy metal were less than the threshold value of the second grade and only the mean content of Pb was more than the value of national background. Results of descriptive statistic showed that the mean content of heavy metals should depend on land utilization and spatial location at some extent. The heavy metal contents were higher in the Southwest and Northwest than in the Central. In addition, the mean contents of Zn and Pb in Dongguan paddy soils were significantly higher than those of Pearl River Delta(PRD) and Taihu Lake region(TLR). Correlation analyses indicated that there existed significant correlation between Cr and Ni in orchard soils, and among Zn, Cd and Cu, between Av-Cu and Cu, between Av-Zn and Cr, Ni, pH value in vegetable soils, and a weak relationship between Cd, Cu and Pb, between Av-Zn and Zn. Principal component analyses(PCA) showed that the order of importance should be Zn > Pb > Cr > Ni > Cu. PMID- 15900720 TI - Adsorption mechanism of phenolic compounds from aqueous solution on hypercrosslinked polymeric adsorbent. AB - The adsorption of two phenols, namely, phenol and salicylic acid(SA) onto a water compatible hypercrosslinked polymeric resin (NJ-8) were studied in terms of pseudo-second-order and first order mechanisms for chemical sorption as well as an intraparticle diffusion mechanism process. Kinetic analysis showed that the intraparticle diffusion process was the essential rate-controlling step. The activation energies of sorption have also been evaluated with the pseudo-second order and intraparticle diffusion constants, respectively. Adsorption equilibrium data were well fitted by the Langmuir, Freundlich and Redlich-Peterson isotherms. Adsorption was exothermic and basically of a type of transition between physical and chemical character. The sorption capacity was higher for SA due to its more hydrophobic. Phenol has a higher adsorption enthalpy since it could form stronger hydrogen bonding on NJ-8. PMID- 15900721 TI - Rapid treatment of atrazine-contaminated water by nickel/iron bimetallic system. AB - The utility of nickel/iron in the remediation of atrazine-contaminated water was investigated. The experimental results showed that nickel/iron had effective catalytic activity in dechlorinating atrazine under acidic conditions. The dechlorination reaction approximately followed the first-order kinetics under the experimental conditions (nickel/iron: 1.0 g/250 ml; C(atrazine) = 20.0 mg/L), the reaction rate increased with decreasing pH value of the reaction solution and increasing the proportion of Ni : Fe within 2.95%. For condition with 2.95% nickel/iron, the reaction rate constants were 0.07518 (R = 0.9927), 0.06212 (R = 0.9846) and 0.00131 min(-1) (R = 0.9565) at pH = 2.0, 3.0 and 4.0, respectively. HPLC analysis was used to monitor the decline of atrazine concentration. PMID- 15900722 TI - Binding of pyrene to dissolved organic matters: fractionation and characterization. AB - Based on a modified Leenheer DOM fractionation scheme, fractionation of DOM from the paddy soil was conducted by using XAD-8 resin into hydrophobic bases (HOB), hydrophobic acids(HOA), acid-insoluble matter (AIM), hydrophobic neutrals (HON) and hydrophilic matter (HIM). In total carbon content of DOM, 35.32% were the HIM and only 0.73% the HOB. However, HOA and AIM altogether occupied 53.45%, while the HON fraction represented 10%. The sorption experiments were conducted to determine the sorption capacity of pyrene on unfractionated DOM and its fractions. Elemental analysis, 1H-NMR and FTIR spectra were carried out on unfractionated DOM and its fractions to examine the relationship between the structure of DOM and partition coefficients (K(oc)). The results showed that HON had a greater affinity for binding pyrene than other fractions. While HON was characterized by large long-chain alkylate (aliphatic structure). AIM exhibited relative higher K(oc) values than HOA and HIM, due to much aromatic structure in AIM, while the high content of carboxylic groups of HOA and HIM depressed their binding capacity. This study demonstrated HON is a key subcomponents of DOM in binding of pyrene, in other words, aliphalic structure in DOM play an important role in binding of pyrene. PMID- 15900723 TI - Effects of nutrients on Microcystis growth more easily forming bloom. AB - Different nutrient media experimentally were N, P and Fe-limited conditions and a serial of diluted BG11 media. The cell change of morphology and life history, cell number, cell color and cell area of Microcystis were analyzed quantitatively. First, the effects of nitrogen, phosphorus and iron depletion were distinctively different. Phosphorus and iron depletion caused more special division cells, slowly growth increasing, the easier change of bigger cell area. Second, the nitrogen and iron depletion could make the color of alga from green to brown. Finally, according to the resource competition and Monod equation, Microcystis kinetics of phosphorus and iron were also examined. K(s) and micro(max) of phosphorus absorption were 0.0352 micromol/L, 0.493 d(-1) respectively; iron absorption: 0.00323 micromol/L, 0.483 d(-1). In a word, some evidences of the Microcystis bloom dominance in certain nutrient conditions were indicated in the experiments. The dominances were determined as the reviving under the adverse circumstances through the special division, the various nitrogen resources, and the lower kinetics of phosphorus and iron than that of most of other algae. The conclusions provided the scientific basis for preventing and managing Microcystis bloom in freshwater. PMID- 15900724 TI - Photocatalytic remediation of gamma-HCH contaminated soil induced by alpha-Fe2O3 and TiO2. AB - Heterogeneous photocatalytic degradation of gamma-HCH on soil surfaces was carried out to evaluate the photocatalytic effectiveness of alpha-Fe2O3 and TiO2 toward degrading gamma-HCH on soil surfaces. After being spiked with gamma-HCH, soil samples were loaded with alpha-Fe2O3 or TiO2 and exposed to UV-light irradiation. Different catalyst loads, 0%, 2%, 5%, 7%, and 10% (wt.) alpha-Fe2O3; 0%, 0.5%, 1%, 2: (wt.)TiO2, were tested for up to 7 d irradiation. The effects of soil thickness, acidity, and humic substances were also investigated. The obtained results indicated that the gamma-HCH photodegradation follows the pseudo first-order kinetics. The addition of alpha-Fe2O3 or TiO2 accelerates the photodegradation of gamma-HCH, while the photodegradation rate decreases when the content of alpha-Fe2O3 exceeds 7% (wt.). The degradation rate increases with the soil pH value. Humic substances inhibit the photocatalytic degradation of gamma HCH. Pentachlorocyclohexene, tetrachlorocyclohexene, and trichlorobenzene are detected as photodegradation intermediates, which are gradually degraded with the photodegradation evolution. PMID- 15900725 TI - Fenton treatment of olive oil mill wastewater--applicability of the method and parameters effects on the degradation process. AB - The low biodegradability of polyphenolic compounds typically found in olive processing indicated that biological treatment is not always successful in the treatment of olive oil mill wastewater in term of COD removal. In this study the results of investigations on the applicability of Fenton's reagent in the treatment of this effluent were discussed. The efficiency of this method was determined. 86% of removal COD was obtained using 5 mol H2O2 and 0.4 mol Fe2+ per liter of crude OMW. The main parameters that govern the complex reactive system, i.e., time, pH, [H2O2] and [Fe(II)] were studied. PMID- 15900726 TI - Analysis of alpha, beta, gamma-hexachlorocyclohexanes in water by novel activated carbon fiber-solid phase microextraction coupled with gas chromatography-mass spectrometry. AB - A fast and simple method for determination of alpha, beta, gamma hexachlorocyclohexanes (HCHs) in water using activated carbon fiber-solid phase microextraction(ACF-SPME) were studied. Results showed the performance of adsorption and desorption of three HCHs on ACF were excellent. A wide linear range from 10 to 100 microg/L and detection limits of the ng/L level were obtained using ACF-SPME with GC-MS in selected ion monitoring(SIM) acquisition mode. The proposed method was also successfully applied for determination of three HCHs in tap water. Compared to commercial fibers, ACF showed some advantages such as better resistance to solvents, higher thermal stability, longer lifetime and lower cost. The data demonstrated that GC-MS with ACF-SPME is well suitable for the analysis of HCHs in water. PMID- 15900727 TI - Use of two-surfactants mixtures to attain specific HLB values for assisted TPH diesel biodegradation. AB - In a surfactant assisted biodegradation process, the choice of surfactant(s) is of crucial importance. The question is: does the type of surfactant (i.e. chemical family) affect the biodegradation process at fixed hidrophillic lypofillic balance (HLB) values? Microcosm assessments were developed using contaminated soil, with around of 5000 mg/kg of hydrocarbons as TPH-diesel. Mixtures of three nonionic surfactants were employed to get a wide range of specific HLB values. Tween20 and Span20 were mixed in the appropriate proportions to get HLB values between 8.6 and 16.7. Tween/Span60 mixtures reached HLB values between 4.7 and 14.9. Finally, Tween/Span80 combinations yielded HLB values between 4.3 and 15. TPH-diesel biodegradation was measured at the beginning, and after 8 weeks, as well as the FCU/gr(soil), as a measure of microorganisms' development during the biodegradation period. A second aim of this work was to assess the use of guar gum as a biodegradation enhancer instead of synthetic products. The conclusions of this work are that surfactant chemical family, and not only the HLB value clearly affects the assisted biodegradation rate. Surfactant's synergism was clearly observed. Regarding the use of guar gum, no biodegradation enhancement was observed for the three assessed concentrations, i.e., 2, 20, and 200 mg/kg, respectively. On the contrary, TPH-diesel removal was lower as the gum concentration increased. It is quite possible that guar gum was used as a microbial substrate. PMID- 15900728 TI - Application of ultrasonic to speciation analysis of heavy metals in soil. AB - In order to reduce the operation time and improve the extraction efficiency, ultrasonic energy by means of ultrasonic bath was used to the modified Tessier sequential extraction for speciation analysis of heavy metals in soil. Extractable contents of Cu, Fe, Mn, Ni, Pb and Zn were measured by atomic absorption spectroscopy (AAS). The merit of the ultrasonic extraction (UE) applied to the modified Tessier method is not only that the operation time for the first 4 fractions was reduced from ca. 18 h to 8 h, comparing with conventional extraction (CE), but also the extraction efficiency was higher. The results for both of UE and CE were consistent. The extractable Cu, Ni and Zn in the sample No. 1 were mainly associated with the third fraction (Fe-Mn oxides fraction), and fourth fraction (organic matter fraction) in the sample No.2. The extractable Fe and Mn were all mainly associated with the third fraction, and Pb the fourth fraction in both of the samples. The effects of concentration of hydroxylamine chloride on the capability for the extraction of studied metals were also studied. PMID- 15900729 TI - Cadmium uptake by different rice genotypes that produce white or dark grains. AB - A pot experiment was conducted to investigate cadmium (Cd) uptake by different rice cultivars that produce white or dark grains. Four cultivars with white grains (hereafter, white rice) and five cultivars with dark colors (hereafter dark rice) were selected for this experiment. Three levels of soil Cd concentrations, background (0), 5 and 10 mg/kg, were used. After harvest, plant biomass, tissue concentrations of Cd, Ca, Fe, Cu and Zn were analyzed. The results showed that Cd concentrations are significantly different between different genotypes, but when comparing the Cd concentrations for the two groups, no significant difference was found. For other divalent cations, Ca concentrations in dark rice were higher than those in white ones (P < 0.001 for shoots, P = 0.037 for roots); Fe concentrations in dark rice were also higher than those in white ones (P = 0.001 either in shoot or root); Zn concentrations in shoot of dark rice were higher than those in white ones, but no significant difference in roots. The total molar concentrations of divalent cations in dark rice were also significantly higher than in white rice. The potential benefit of higher Ca and Fe concentrations in dark rice and similar Cd concentrations in both groups is also discussed in this paper. PMID- 15900730 TI - Aerobic dechlorination of cis- and trans-dichloroethenes by some indigenous bacteria isolated from contaminated sites in Africa. AB - The innate toxicity of dichloroethenes (DCEs) and their tendency to be reduced to vinyl chloride (VC) (a known human carcinogen) is a cause for environmental concern. Aerobic bacteria capable of growth on cis- and trans-DCEs as sole carbon and energy sources were isolated by enrichment culture technique and identified to belong to the genera; Bacillus, Pseudomonas and Acinetobacter. Axenic and mixed cultures of the bacterial isolates utilized DCEs at concentrations above the maximum contaminant level allowable in drinking water by the Environmental Protection Agency. Their specific growth rate constant ranged significantly (P < 0.05) between 0.346-0.552 and 0.461-0.667 d(-1); while the maximum specific substrate utilization rate ranged significantly (P < 0.05) between 20.01-29.79 and 31.40-42.83 nmol h(-1) (mg of protein)(-1) in cis- and trans-DCE, respectively. The optimum growth was observed at 30 degrees C and at a pH of 7.0 with up to 96% of the stoichiometric-expected chloride released. Serial adaptation positively affected the growth yields and dehalogenase activities of the organisms with multiple antibiotic patterns also demonstrated by the isolates. These findings therefore indicated the important roles that these organisms may play in the bioremediation of sites polluted with chlorinated ethene compounds in Africa. PMID- 15900731 TI - Catalytic oxidation of calcium sulfite in solution/aqueous slurry. AB - Forced oxidation of calcium sulfite aqueous slurry is a key step for the calcium based flue gas desulfurization (FGD) residue. Experiments were conducted in a semi-batch system and a continuous flow system on lab scales. The main reactor in semi-batch system is a 1000 ml volume flask. It has five necks for continuous feeding of gas and a batch of calcium sulfite solution/aqueous slurry. In continuous flow system, the main part is a jacketed Pyrex glass reactor in which gas and solution/aqueous slurry are fed continuously. Calcium sulfite oxidation is a series of complex free-radical reactions. According to experimental results and literature data, the reactions are influenced significantly by manganese as catalyst. At low concentration of manganese and calcium sulfite, the reaction rate is dependent on 1.5 order of sulfite concentration, 0.5 order of manganese concentration, and zero order of oxygen concentration in which the oxidation is controlled by chemical kinetics. With concentrations of calcium sulfite and manganese increasing, the reactions are independent gradually on the constituents in solution but are impacted by oxygen concentration. Manganese can accelerate the free-radical reactions, and then enhances the mass transfer of oxygen from gas to liquid. The critical concentration of calcium sulfite is 0.007 mol/L, manganese is 10(-4) nol/L, and oxygen is of 0.2-0.4 atm. PMID- 15900732 TI - Influence of the Haizhou Open Pit Coal Mine on the atmospheric flow over Fuxin, China. AB - The influence of the Haizhou Open Pit Mine on the atmospheric flow in nearby Fuxin City in China was analyzed with the aid of the steady-state Navier-Stokes equations. The finite element method was used to obtain numerical solutions to these equations. The results showed that the Haizhou Open Pit Coal Mine contributes to the turbulent flow in the Fuxin City and its surroundings. However, when compared with the climatic effects, the open pit mine has a relatively small impact on the atmospheric flow over Fuxin. PMID- 15900733 TI - Assessment on the pollution of nitrogen and phosphorus of Beijing surface water based on GIS system and multivariate statistical approaches. AB - This paper presented the characteristics of nitrogen and phosphorus pollution in Beijing surface water during the survey. A significant difference was found out in concentration distribution of various parameters of nitrogen and phosphorus. Most water bodies in five water systems were polluted by total nitrogen with the content even up to 120 mg/L which was higher than exceeded the fifth class standard of national surface water quality standard GB3838-2002 except for several segments of Chaobaihe and Yongdinghe. Ammonia and phosphorus showed a similar tendency of distribution with higher content in Daqinghe, Beiyunhe and Jiyunhe water systems, but with relatively low concentrations in Chaobaihe and Yongdinghe water systems. Meanwhile, nitrate was found at comparatively low content (mostly less than 10 mg/L) and could fit for corresponding water quality requirements. Totally, the water quality of Daqinghe, Jiyunhe and Beiyunhe river systems as well as the lower reaches of Yongdinghe and Chaobaihe was contaminated seriously with high content of total nitrogen and phosphorus. Through multivariate statistical approaches, it can be concluded that total nitrogen, ammonia and total phosphorus was highly correlated to chemical oxygen demand, biochemical oxygen demand, dissolved oxygen and electrical conductivity, which explained the same pollution source from anthropogenic activities. PMID- 15900734 TI - Microbial changes in rhizospheric soils contaminated with petroleum hydrocarbons after bioremediation. AB - Effects of bioremediation on microbial communities in soils contaminated with petroleum hydrocarbons are a scientific problem to be solved. Changes in dominate microbial species and the total amount of microorganisms including bacteria and fungi in rhizospheric soils after bioremediation were thus evaluated using field bioremediation experiments. The results showed that there were changed dominant microorganisms including 11 bacterial strains which are mostly Gram positive bacteria and 6 fungal species which were identified. The total amount of microorganisms including bacteria and fungi increased after bioremediation of microbial agents combined with planting maize. On the contrary, fungi in rhizospheric soils were inhibited by adding microbial agents combined with planting soybean. PMID- 15900735 TI - Treatment of anthraquinone dye wastewater by hydrolytic acidification-aerobic process. AB - Experiment on microbial degradation with two kinds of biological process, hydrolytic acidification-aerobic process and aerobic process was conducted to treat the anthraquinone dye wastewater with COD(Cr) concentration of 400 mg/L and chroma 800. The experimental result demonstrated that the hydrolytic-aerobic process could raise the biodegradability of anthraquinone dye wastewater effectively. The effluent COD(Cr) can reach 120-170 mg/L and chroma 150 which is superior to that from simple aerobic process. PMID- 15900736 TI - Projection pursuit cluster model and its application in water quality assessment. AB - One of the difficulties frequently encountered in water quality assessment is that there are many factors and they cannot be assessed according to one factor, all the effect factors associated with water quality must be used. In order to overcome this issues the projection pursuit principle is introduced into water quality assessment, and projection pursuit cluster (PPC) model is developed in this study. The PPC model makes the transition from high dimension to one dimension. In other words, based on the PPC model, multifactor problem can be converted to one factor problem. The application of PPC model can be divided into four parts: (1) to estimate projection index function Q(a -->); (2) to find the right projection direction a -->; (3) to calculate projection characteristic value of the i th sample z(i), and (4) to draw comprehensive analysis on the basis of z(i). On the other hand, the empirical formula of cutoff radius R is developed, which is benefit for the model to be used in practice. Finally, a case study of water quality assessment is proposed in this paper. The results showed that the PPC model is reasonable, and it is more objective and less subjective in water quality assessment. It is a new method for multivariate problem comprehensive analysis. PMID- 15900737 TI - Modification on the conventional procedure to measure AOC in drinking water. AB - Additional phosphorus will be introduced to water sample if the conventional procedure is used to measure assimilable organic carbon (AOC) in drinking water. It has been shown that there are the cases that phosphorus is the limiting nutrient for microbial growth in drinking water. The measured value of AOC would not be able to indicate appropriately the regrowth potential of bacteria in this case. The conventional procedure used to measure AOC was modified to avoid the introduction of additional phosphorus to water sample in this study. It was shown that it was feasible to measure AOC in water using the modified procedure. Furthermore, the measured value of AOC determined by the modified procedure could indicate appropriately the regrowth potential of bacteria in drinking water despite either organics or phosphorus was the limiting nutrient for bacterial regrowth. PMID- 15900738 TI - Ammonium removal by modified zeolite from municipal wastewater. AB - Ammonium removal by modified zeolite, H-form and Na-form zeolite, were examined by batch-type methods. The adsorption of ammonium on modified zeolite was exothermic process. The saturation adsorption capacity of ammonium on H-form and Na-form zeolite were 21.23 and 41.15 mg/g, respectively. After ten times adsorption-desorption-readsorption cycles the standard deviations of H-form and Na-form zeolite were 6.34% and 6.59%. The zeolite adsorption process has proved cost effective and practical in reducing ammonium by H-form and Na-form zeolite in municipal wastewater from concentration 27.68 mg/L to 2.80 mg/L and 5.91 mg/L. PMID- 15900739 TI - Model-based evaluation on the conversion ratio of ammonium to nitrite in a nitritation process for ammonium-rich wastewater treatment. AB - Modeling for nitritation process was discussed and analyzed quantitatively for the factors that influence nitrite accumulation. The results indicated that pH, inorganic carbon source and Hydraulic Retention Time (HRT) as well as biomass concentration are the main factors that influenced the conversion ratio of ammonium to nitrite. A constant high pH can lead to a high nitritation rate and results in high conversion ratio on condition that free ammonia inhibition do not happen. In a CSTR system, without pH control, this conversion ratio can be monitored by pH variation in the reactor. The pH goes down far from the inlet level means a strongly nitrite accumulation. High concentration of alkalinity can promoted the conversion ratio by means of accelerating the nitritation rate through providing sufficient inorganic carbon source(carbon dioxide). When inorganic carbon source was depleted, the nitritation process stopped. HRT adjustment could be an efficient way to make the nitritation system run more flexible, which to some extent can meet the requirements of the fluctuant of inlet parameters such as ammonium concentration, pH, and temperature and so on. Biomass concentration is the key point, especially for a CSTR system in steady state, which was normally circumscribed by the characteristics of bacteria and may also affected by aeration mode and can be increased by prolonging the HRT on the condition of no nitrate accumulation when no recirculation available. The higher the biomass concentration is, the better the nitrite accumulation can be obtained. PMID- 15900740 TI - Desulfurization of dibenzothiophene by a newly isolated Corynebacterium sp. ZD-1 in aqueous phase. AB - Sulfur emission through fuel combustion is a global problem because it is a major cause of acid rain. Crud oil contains many heterocyclic organic sulfur compounds, among which dibenzothiophene (DBT) and DBTs bearing alkyl substitutions usually are representative compounds. A strain was isolated from refinery sludge and identified as Corynebacterium ZD-1. The behavior of DBT degradation by ZD-1 in aqueous phase was investigated. Corynebacterium ZD-1 could metabolize DBT to 2 hydroxybiphenyl(2-HBP) as the dead-end metabolite through a sulfur-specific pathway. In shake flask culture, ZD-1 had its maximal desulfurization activity in the late exponential growth phase and the specific production rate of 2-HBP was about 0.14 (mmol x kg dry cell(-1) x min(-1), mmol x KDC(-1) x min(-1)). Active resting cells for desulfurization should be prepared only in this period. 2-HBP inhibited the growth of strain ZD-1, the production of DBT degradation enzymes, and the activity of enzymes. Sulfate inhibited the production of dibenzothiophene (DBT) degradation enzymes but had no effect on the enzymes' activity. The production rates of 2-HBP at lower cell densities were higher and the maximum amount conversion of DBT to 2-HBP (0.067 mmol/L) after 8 h was gained at 9.2 g dry cell/L rather higher cell density. The results indicated that this newly isolated strain could be a promising biocatalyst for DBT desulfurization. PMID- 15900741 TI - Intensification of adsorption process by using the pyrolytic char from waste tires to remove chromium (VI) from wastewater. AB - Pyrolysis has the potential of transforming waste into valuable recyclable products. Pyrolytic char (PC) is one of the most important products from the pyrolysis of used tires. One of the most significant applications for pyrolytic char recovered is used for the removal of Cr(VI) in the wastewater effluent to control waste by waste. The surface chemistry properties of surface element distribution/concentration and chemical structure were examined for the pyrolytic char and the commercial activated carbon (CAC) respectively. The results showed that surfaces of PC possesses a large amount of ester and hydrocarbon graft, whereas there are mainly carbon functional components of C-OH, C=O and COOH on the surface of CAC. Therefore the surface electronegativity of PC is lower than that of CAC in the water. The repulsive interactions between the surfaces of PC and the negatively charged Cr(VI) ion are weaker than that of CAC, which results in an intensification of the adsorption process by the utilization of PC. The adsorption isotherms of Cr(VI) ion on the two kinds of carbons were determined experimentally. The larger adsorption amount on the PC in the case of Cr(VI) may be attributed mainly to its special surface micro-chemical environment. The mechanism of the removal Cr(VI) from aqueous solution was assumed to be the integration of adsorption and redox reaction. The adsorption was the rate controlled step for Cr(VI) removal. The adsorption of Cr(VI) was identified as pseudo-second-order kinetics. The rate constants of adsorption were evaluated. PMID- 15900742 TI - Modelling the biological performance of a side-stream membrane bioreactor using ASM1. AB - Membrane bioreactors (MBRs) are attracting global interest but the mathematical modeling of the biological performance of MBRs remains very limited. This study focuses on the modelling of a side-stream MBR system using Activated Sludge Model No. 1 (ASM1), and comparing the results with the modelling of traditional activated sludge processes. ASM1 parameters relevant for the long-term biological behaviour in MBR systems were calibrated (i.e. Y(H) = 0.72gCOD/gCOD, Y(A) = 0.25gCOD/gN, b(H) = 0.25 d(-1), b(A) = 0.080 d(-1) and f(P) = 0.06), and generally agreed with the parameters in traditional activated sludge processes, with the exception that a higher autotrophic biomass decay rate was observed in the MBR. A sensitivity analysis for steady state operation and DO dynamics suggested that the biological performance of the MBR system (the sludge concentration, effluent quality and the DO dynamics) are very sensitive to the parameters(i.e. Y(H), Y(A), b(H), b(A), micro(maxH) and micro(maxA)), and influent wastewater components(X(I), S(S), X(S), S(NH)). PMID- 15900743 TI - Biogenic VOCs emission inventory development of temperate grassland vegetation in Xilin River basin, Inner Mongolia, China. AB - Given the key role of biogenic volatile organic compounds (VOCs) to tropospheric chemistry and regional air quality, it is important to generate accurate VOCs emission inventories. However, only a less fraction of plant species, in temperate grassland of Inner Mongolia, has been characterized by quantitative measurements. A taxonomic methodology, which assigns VOCs measurements to unmeasured species, is an applicable and inexpensive alternation for extensive VOCs emission survey, although data are needed for additional plant families and genera to further validate the taxonomic approach in grassland vegetation. In this experiment, VOCs emission rates of 178 plant species were measured with a portable photoionization detector (PID). The results showed the most of genera and some families have consistent feature of their VOCs emission, especially for isoprene, and provide the basic premise of taxonomic methodology to develop VOCs emission inventories for temperate grassland. Then, the taxonomic methodology was introduced into assigning emission rate to other 96 species, which no measured emission rates available here. A systematical emission inventory of temperate grassland vegetation in Inner Mongolia was provided and further evidence that taxonomy relationship can serve as a useful guide for generalizing the emissions behavior of many, but not all, plant families and genera to grassland vegetation. PMID- 15900744 TI - Volatilization behaviors of diesel oil from the soils. AB - The volatilization of diesel oil, Shengli crude oil and 90 # gasoline on glass surface of petri dishes were conducted at the ambient temperature of 25 degrees C. Diesel oil evaporates in a power manner, where the loss of mass is approximately power with time. 90 # gasoline evaporates in a logarithmic with time. Where as the volatilization of Shengli crude oil fit either the logarithmic or power equation after different time, and has similar R2. And the effects of soil type and diesel oil and water content on volatilization behavior in unsaturated soil were studied in this paper. Diesel oil and water content in the soils play a large role in volatilization from soils. Appropriate water helps the wicking action but too much water stops it. The wicking action behaves differently in four different types of soils in the same volatilization experiment of 18% diesel oil content and air-dry condition. PMID- 15900745 TI - Biological nitrogen removal with enhanced phosphate uptake in (AO)2 SBR using single sludge system. AB - Simultaneous biological phosphorus and nitrogen removal with enhanced anoxic phosphate uptake via nitrite was investigated in an anaerobic-aerobic-anoxic aerobic sequencing batch reactor ((AO)2 SBR). The system showed stable phosphorus and nitrogen removal performance, and average removals for COD, TN and TP were 90%, 91% and 96%, respectively. The conditions of pH 7.5-8.0 and temperature 32 degrees C were found detrimental to nitrite oxidation bacteria but favorable to ammonia oxidizers, and the corresponding specific oxygen uptake rates(SOUR) for phase 1 and 2 of nitrification process were 0.7 and 15 mgO2/(gVSS x h) in respect, which led to the nitrite accumulation in aerobic phase of (AO)2 SBR. Respiratory tests showed that 40 mgNO2-N/L did not deteriorate the sludge activity drastically, and it implied that exposure of sludge to nitrite periodically enabled the biomass to have more tolerance capacity to resist the restraining effects from nitrite. In addition, batch tests were carried out and verified that denitrifying phosphorus accumulation organisms (DPAOs) could be enriched in a single sludge system coexisting with nitrifiers by introducing an anoxic phase in an anaerobic-aerobic SBR, and the ratio of the anoxic phosphate uptake capacity to aerobic phosphate uptake capacity was 45%. It was also found that nitrite (up to 20 mgNO2-N/L) was not inhibitory to anoxic phosphate uptake and could serve as an electron acceptor like nitrate, but presented poorer efficiency compared with nitrate. PMID- 15900746 TI - Anammox transited from denitrification in upflow biofilm reactor. AB - Anammox was successfully transited from heterotrophic denitrification and autotrophic denitrification in two upflow biofilm reactors, respectively. The results showed that the volumetric loading rate and nitrogen removal efficiency in the reactor transited from heterotrophic denitrification were higher than that in its counterpart. When the hydraulic retention time was 12 h or so, the total nitrogen loading rate was about 0.609 kg N/(m3 x d), and the effluent ammonia and nitrite concentrations were less than 8.5 mg/L and 2.5 mg/L, respectively. The upflow anammox biofilm reactor was capable of keeping and accumulating the slow growing bacteria efficiently. During operation of the reactor, the biomass color was gradually turned from brownish to red, and the ratio of ammonia consumption, nitrite consumption and nitrate production approached the theoretical one. These changes could be used as an indicator for working state of the reactor. PMID- 15900747 TI - Use of rural energy resources and eco-environmental degradation in Tibet. AB - Cattle dung, firewood, and crop straw have being used as survival necessities by farmers and herdsmen for thousands of years in Tibet. Until recently such biotic energy source still constitutes more than 92 per cent of total rural energy consumption due to lack of petroleum, coal and new alternative energy sources. As a result, environmental degradation such as land desertification, soil erosion, grassland degradation and soil fertility reduction is increasingly aggravated, the area of desertified land has increased 1467.5 km2 from 1991 to 1997. Degraded area of grassland has reached 2.60 x 10(7) hm2, increased by 116.1% from 1987 to 1996. To prevent further deterioration of eco-environment in Tibet great efforts should be made to make full use of ample solar energy, wind energy and other biotic energy of the Qinghai-Tibet Plateau. The solar cooking stoves and solar hothouse, expand forest area and replace existing abiotic energy sources with firewood forest should be popularized. This is an urgent task to protect the eco environment of Tibet today. PMID- 15900748 TI - Organic carbon stock in topsoil of Jiangsu Province, China, and the recent trend of carbon sequestration. AB - Data collection of soil organic carbon(SOC) of 154 soil series of Jiangsu, China from the second provincial soil survey and of recent changes in SOC from a number of field pilot experiments across the province were collected. Statistical analysis of SOC contents and soil properties related to organic carbon storage were performed. The provincial total topsoil SOC stock was estimated to be 0.1 Pg with an extended pool of 0.4 Pg taking soil depth of 1 m, being relatively small compared to its total land area of 101700 km2 . One quarter of this topsoil stock was found in the soils of the Taihu Lake region that occupied 1/6 of the provincial arable area. Paddy soils accounted for over 50% of this stock in terms of SOC distribution among the soil types in the province. Experimental data from experimental farms widely distributed in the province showed that SOC storage increased consistently over the last 20 years despite a previously reported decreasing tendency during the period between 1950--1970. The evidence indicated that agricultural management practices such as irrigation, straw return and rotation of upland crops with rice or wheat crops contributed significantly to the increase in SOC storage. The annual carbon sequestration rate in the soils was in the range of 0.3-3.5 tC/(hm2 x a), depending on cropping systems and other agricultural practices. Thus, the agricultural production in the province, despite the high input, could serve as one of the practical methods to mitigate the increasing air CO2. PMID- 15900749 TI - Investigation of natural VOC emitted from tropical vegetations in China. AB - Twenty-three kinds of typical plants in Xishuangbanna, the tropical area of southwestern China, were screened to estimate the emission rates of isoprene and monoterpenes by adopting bag-enclosure and curette sampling methods followed by a GC-FID analysis. It was found that the Ficus species were mainly emitting isoprene and most tropical vegetations were mainly releasing monoterpenes. The results also showed that the emissions of isoprene were affected by both temperature and PAR(Photosynthetic Active Radiation), while monoterpene emissions were mainly temperature-dependent. PMID- 15900750 TI - Phytoremediation for phenanthrene and pyrene contaminated soils. AB - Phytoremediation of soil contaminated with phenanthrene and pyrene was investigated using twelve plant species. Plant uptake and accumulation of these chemicals were evaluated. At the end of the experiment(45 d), the remaining respective concentrations of soil phenanthrene and pyrene in spiked vegetated soils, with initial phenanthrene of 133.3 mg/kg and pyrene of 171.5 mg/kg, were 8.71-16.4 and 44.9--65.0 mg/kg, generally 4.7%--49.4% and 7.1%--35.9% lower than their concentrations in the non-vegetated soils. The loss of phenanthrene and pyrene in vegetated spiked soils were 88.2%--93.0% and 62.3%--73.8% of the added amounts of these contaminants, respectively. Although plant uptake and accumulation of these compounds were evident, and root concentrations and RCFs (root concentration factors; defined as the ratio of PAH concentrations in roots and in the soils on a dry weight basis) of these compounds significantly positively correlated to root lipid contents, plant uptake and accumulation only accounted for less than 0.01% and 0.23% of the enhanced loss of these chemicals in vegetated versus non-vegetated soils. In contrast, plant-promoted microbial biodegradation was the dominant mechanism of the phytoremediation for soil phenanthrene and pyrene contamination. Results from this study suggested a feasibility of the establishment of phytoremediation for soil PAH contamination. PMID- 15900751 TI - Synergistic effect of palladium and oxygen vacancies in the Pd/perovskite catalysts synthesized by the spc method. AB - A series of oxygen-deficient perovskite-supported palladium catalysts were prepared by the "solid phase crystallization" (spc) method and investigated with XRD, TPR, TPD, TEM, XPS, BET analysis and CO oxidation. It was found that Pd/perovskite catalysts synthesized by the spc method were more active for CO oxidation than the calcined LaCo0.95 Pd0.05 O3, where Pd dispersed in the solid solution. H2-reducing treatment in the spc method could yield not only high dispersed fine Pd particles on the perovskite surface but also oxygen-deficient structure. In these perovskite-supported Pd catalysts, oxygen vacancies adsorbed, activated and supplied oxygen to the active Pd sites, where the oxidation occurred with adsorbed CO. The high activities were due to the cooperative action of Pd and oxygen vacancies. PMID- 15900752 TI - Absorption of phosphorus from wastewater by aged refuse excavated from municipal solid waste landfill. AB - Municipal solid waste(refuse) landfill stabilizes as the refuse degrades. After years of biodegradation, the refuse in the landfill becomes stabilized and aged, which may vary with the local climate, humidity, and composition of refuse placed. In this work, it is found that the refuse with an age of over 8 years at Shanghai Refuse Landfill has been significantly stabilized and sufficiently aged and is thus suitable for excavation. The 8-year old aged refuse is mechanically screened, and the fine fractions of refuse(aged refuse) with a diameter less than 2 cm are then used as a biological absorbent for removal of both inorganic and organic phosphorus in livestock wastewater and prepared aqueous solution. It is proved that the aged refuse is very effective for the quantitative removal of both types of phosphorus. The absorption mechanism is proposed. It is considered that phosphorus is firstly absorbed onto the surface of the aged refuse and then used as a substrate for the growth of microorganisms which ultimately leave the aged refuse as sludge. PMID- 15900753 TI - Adsorption of Pb, Cd to Fe, Mn oxides in natural freshwater surface coatings developed in different seasons. AB - Metal oxides( Fe, Mn oxides) in natural surface coatings(biofilms and associated minerals) are believed to play a significant role in the fate and transport of trace metal in aquatic environments. Seasonal variation of Fe, Mn oxides and organic materials in surface coatings, which were developed periodically on glass slides in Nanhu Lake, Jilin Province, China over the time frame of three seasons, was investigated in order to understand the influence of metal oxides on Pb and Cd adsorption to heterogeneous surface coating materials (biofilm). Pb and Cd adsorption was measured under controlled laboratory conditions(mineral salts solution with defined speciation, ionic strength 0.05 mol/L, 25 degrees C and pH 6.0). The classical Langmuir adsorption isotherm was applied to estimate equilibrium coefficients of Pb and Cd adsorption to the surface coatings. In general, components in the surface coatings varied greatly with seasons altering and obtained higher concentrations in summer while the content of iron oxides always exceeded that of manganese oxides. Correlation analyses between the maximum adsorption of Pb and Cd and components in the surface coatings developed periodically indicated that Pb phase association with Mn oxides and Cd phase association with Fe oxides as well as Mn oxides were statistically significant. Effect of Mn oxides on Cd adsorption was confirmed in view of its higher content in the surface coatings. The importance of ferromanganese oxides for Pb and Cd adsorption to the natural surface coatings developed in different seasons was evidenced. PMID- 15900754 TI - Effects of cracks and some key factors on emissions of nitrous oxide in paddy fields. AB - Paddy field is a primary agricultural landscape in the south of China and is often regarded as one of main sources emitting nitrous oxide to atmosphere. The nitrous oxide emissions under a variety of paddy field practices, such as fertilization, flooding/draining management were investigated to study on agricultural activities on paddy field affect the dynamic process of the emission. Under no addition of fertilizers the average emission flux of nitrous oxide was 8.55 microg/(m2 x h) during the rice( Oryza Sativa L. ) growth season. The results indicated that most of nitrous oxide emissions occurred during the crack forming-and-expansion period when paddy field was being drained. The diurnal emissions peak of nitrous oxide appeared at 20:30 at night in cracked rice fields. The statistical analysis suggested that the correlation of nitrous oxide emissions flux ( Y) with soil water content ( X1 ), soil temperature ( X2 ), and Eh ( X3 ), could be described in a regression equation: Y= - 1498.95 + 2895.48X, + 50.63 X2 - 96.99X1 x X2 + 0.006X2 x X3. There were the different power equations to simulate the correlations between the everyday dynamic N2O emissions and the mean surface area of cracks, mean volume and depth of cracks respectively during paddy soil drying by soil columns incubation experiments. Taken all together, the current study presented a dynamic analysis of nitrous oxide emission of paddy field under various conditions, therefore provided a basis for the management to balance between environmental effect and paddy field activities. PMID- 15900755 TI - Interaction of humic substances and hematite: FTIR study. AB - The present work extended the knowledge on the binding and complexation of humic substances( humic acid or fulvic acid) and hematite by Fourier transform infrared spectroscopy( FTIR) . The FTIR data gained gave the consist evidences by two different sampling preparation methods that the interaction mechanism between humic substances and hematite was mainly conform to the ligand-exchange involving carboxylic functional groups of humic substances and the surfaces sites of hematite. The present method, although associated with some uncertainties, provided an opportunity to increase the knowledge in this field. PMID- 15900756 TI - Distribution, sources and potential toxicological significance of polycyclic aromatic hydrocarbons(PAHs) in Guanting Reservoir sediments, China. AB - The Guanting Reservoir lost its function as the second biggest drinking water source for Beijing due to the pollutions from the upstream flow of Yongding River in 1997. From 1998, lots of studies were carried out to renew the function of Guanting Reservoir as domestic drinking water source before 2008 Olympic Games. This is the first time that polycyclic aromatic hydrocarbon concentrations in the surface sediment of Guanting Reservoir have been analyzed. A distinctive spatial distribution of PAHs was observed. Sediments from four sites along Inlet of Yongding River to reservoir bam had PAHs concentrations of 1377-2855 microg/g dw in descending order. The composition of PAHs is investigated and used to assess petrogenic, combustion and naturally derived of the sediments samples of Guanting Reservoir. Special PAHs ratios, such as phenanthrene/anthracene (P/A) and fluoranthene/pyrene (FI/Pyr) were calculated to assess the relative importance of different origins. The data confirmed a relatively high level of petrogenic contamination in four sites. These high PAHs levels were associated with the input of untreated and partially treated industrial sewage. In addition, the concentrations of PAHs compounds of samples indicated that sediments of reservoir were most likely to pose potential biological impairment. PMID- 15900757 TI - Spatial variations in daily average CO2 concentrations above wetland surface of Xianghai National Nature Reserve, China. AB - Horizontal and vertical variations of daily average CO2 concentration above the wetland surface were studied in Xianghai National Nature Reserve of China in August, 2000. The primary purpose was to study spatial distribution characteristics of CO2 concentration on the four levels of height(0.1 m, 0.6 m, 1.2 m and 2 m) and compare the differences of CO2 concentration under different land covers. Results showed that daily average CO2 concentration above wetland surface in Xianghai National Natural Reserve was lower than that above other wetlands in northeast China as well as the worldwide average, suggesting that Xianghai wetland absorbed CO2 in August and acted as "sink" of CO2. The horizontal variations on the four levels of height along the latitude were distinct, and had the changing tendency of "decreasing after increasing" with the increase of height. The areas with obvious variations were consistent on different levels of height, and those with the highest variations appeared above surface of shore, sloping field, Typha wetland and Phragmites wetland; the vertical variations were greatly different, with the higher variations in Phragmites wetland and Typha wetland, and the lands near the shore and the sloping field with the lower variations. Spatial variations of daily average CO2 concentrations above wetland surface were affected by surface qualities and land covers. PMID- 15900758 TI - Synchronous municipal sewerage-sludge stabilization. AB - A study on a pilot plant accomplishing synchronous municipal sewerage-sludge stabilization was conducted at a municipal sewerage treatment plant. Stabilization of sewerage and sludge is achieved in three-step process: anaerobic reactor, roughing filter and a microbial-earthworm-ecofilter. The integrated ecofilter utilizes an artificial ecosystem to degrade and stabilize the sewerage and sludge. When the hydraulic retention time(HRT) of the anaerobic reactor is 6 h, the hydraulic load(HL) of the bio-filter is 16 m3/(m2 x d), the HL of the eco filter is 5 m3/(m2 x d), the recycle ratio of nitrified liquor is 1.5, the removal efficiency is 83%-89% for COD(Cr), 94%-96% for BOD5, 96%-98% for SS, and 76%-95% for NH3-N. The whole system realizes the zero emission of sludge, and has the characteristics of saving energy consumption and operational costs. PMID- 15900759 TI - Designing principles of an ecological water storage basin on coastal saline: a case study. AB - The degradation of water source environment becomes serious problems accompanying with rapid urbanization in China. Ecological engineering provides ecologically sound and cost-effective solution to solving this problem. As a case study, a 15 hm2 ecological water storage basin for a water plant was designed and constructed on the TEDA area in Tianjin City. Located on saline, the construction of this project has to face serious difficulties, such as high salinity, scarce seed banks of macrophytes, and strong winds. Freshwater replacement, soil amendation and macrophytes planting at the basinshore, wooden water breaker and plastic membrane installation and other measures were conducted for the assistance of plant community establishment. The result showed that the chloride concentration in the basin water decreased from 11600 mg/L to less than 100 mg/L, and the chloride content in the basin sediment decreased from 2.1 % to 0.35% after freshwater soaking. The introduced macrophytes of 8 species all survived and 11 other macrophytes species were occurred in the basin. A new ecosystem was created with increased biological diversity in the original saline, and the water quality was improved. This ecological water storage basin also provided a pleasing landscape for local people. PMID- 15900760 TI - Effects of methyl-CD and humic acid on hydrolytic degradation of the herbicide diclofop-methyl. AB - Hydrolytic degradation of the herbicide diclofop-methyl was investigated in the multi-pH deionized water, natural aquatic systems and soil suspensions. Resulting data indicated that the herbicide was stable in the acidic and nearly neutral solutions for at least 15 d. The herbicide diclofop-methyl rapidly dissipated in the natural aquatic systems and soil suspensions with half-lives less than 4 d. Methyl-CD (partially methylated beta-cyclodextrin) improved its hydrolytic degradation in the pH 8 deionized water and natural aquatic systems while humic acid inhibited its hydrolytic degradation at the same conditions. But dissolved organic matter in the natural aquatic systems and soil suspensions increased its hydrolysis. Two catalysis mechanisms were introduced to describe the effects of cyclodextrin and organic matter on its hydrolytic metabolism. Though inorganic ions maybe improved its hydrolysis reaction in the natural aquatic systems, Fe2+ and Cu2+ did not form complexes with the herbicide and had poor influences on its hydrolytic degradation whether cyclodextrin was added or not. PMID- 15900761 TI - Effect of free-air CO2 enrichment on nematode communities in a Chinese farmland ecosystem. AB - At a rice-wheat rotational free-air CO2 enrichment( FACE) platform, the effect of elevated atmospheric CO2 on soil nematode communities in a farmland ecosystem was studied. Wheat plots were exposed to elevated atmospheric CO2 (ambient 370 microl/L + 200 microl/L). 32 families and 40 genera of nematode were observed in soil suspensions during the study period. Under FACE treatment, the numbers of total nematodes, bacterivores and fungivores exhibited an increasing trend. Because of the seasonal variation of soil temperature and moisture, the effect of elevated atmospheric CO2 on soil nematodes was only observed under favorable conditions. The response of nematode communities to elevated atmospheric CO2 may indicate the change of soil food web. PMID- 15900762 TI - Visible light induced photodegradation of organic pollutants on nitrogen and fluorine co-doped TiO2 photocatalyst. AB - The nitrogen and fluorine co-doped TiO2 polycrystalline powder was synthesized by calcinations of the hydrolysis product of tetrabutyl titanate with ammonium fluoride. Nitrogen and fluorine co-doping causes the absorption edge of TiO2 to shift to a lower energy region. The photocatalytic activity of co-doped TiO2 with anatase phases was found to be 2.4 times higher than that of the commercial TiO2 photocatalyst Degussa P25 for phenol decomposition under visible light irradiation. The co-doped TiO2 powders only contain anatase phases even at 1000 degrees C. Apparently, ammonium fluoride added retarded phase transformation of the TiO2 powders from anatase to rutile. The substitutional fluorine and interstitial nitrogen atoms in co-doped TiO2 polycrystalline powder were responsible for the vis light response and caused the absorption edge of TiO2 to shift to a lower energy region. PMID- 15900763 TI - Role of nitrification inhibitor DMPP (3, 4-dimethylpyrazole phosphate) in NO(3-) N accumulation in greengrocery( Brassica campestris L. ssp. chinensis) and vegetable soil. AB - The influence of nitrification inhibitor (NI) 3, 4-dimethylpyrazole phosphate (DMPP) on nitrate accumulation in greengrocery (Brassica campestris L. ssp. chinensis) and vegetable soil at surface layer were investigated in field experiments in 2002 and 2003. Results showed that NI DMPP took no significant effect on yields of edible parts of greengrocery, but it could significantly decrease NO(3-)-N concentration in greengrocery and in vegetable soil at surface layer. In addition, NI DMPP could reduce the NO(3-)-N concentration during the prophase stage of storage. PMID- 15900764 TI - DNA damage of germ cell of rat induced by nitrotoluene chemicals. AB - 2,4-dinitrotoluene(2,4-DNT), 2,6-dinitrotoluene(2,6-DNT) and 4-nitrotoluene(4-NT) are typical pollutants in the Songhua River of Northeast China. Sertoli/germ cell cocultures and single cell gel electrophoresis(SCGE) are applied to investigate whether they have genotoxicity on DNA damage of germ cell of Kunming male rat. The results showed that all three nitrotoluene compounds tested could induce DNA single-strand breaks of the germ cell. A significant relationship is found between logarithm dose and the degree of DNA damage, which implies that 2,4-DNT, 2,6-DNT and 4-NT have genotoxicity and can induce the germ cell DNA strand to break in vitro. PMID- 15900765 TI - Investigation on activated semi-coke desulfurization. AB - An activated semi-coke with industrial-scale size was prepared by high-pressure hydrothermal chemistry activation, HNO3 oxidation and calcination activation in proper order from Inner Mongolia Zhalainuoer semi-coke, which is rich in resource and cheap in sale. SO2 adsorption capacity on this activated semi-coke was assessed in the fixed bed in the temperature range of 60-170 degrees C, space velocity range of 500-1300 h(-1), SO2 concentration of 1000-3000 ppmv, and N2 as balance. The surface area, elemental and proximate analysis for both raw semi coke and activated semi-cokes were measured. The experimental results showed that the activated semi-coke has a high adsorption capacity for sulfur dioxide than the untreated semi-coke. This may be the result of increase of surface area on activated semi-coke and surface oxygen functional groups with basicity characteristics. Comparison to result of FTIR, it is known that group of -C-O-C- may be active center of SO2 catalytic adsorption on activated semi-coke. PMID- 15900766 TI - Evolution of nutrient structure and phytoplankton composition in the Jiaozhou Bay ecosystem. AB - The inventories of nutrients in the surface water and large phytoplankton( > 69 microm) were analyzed from the data set of JERS ecological database about a typical coastal waters, the Jiaozhou Bay, China, from 1960s for N, P and from 1980s for Si. By examining long-term changes of nutrient concentration, calculating stoichiometric balance, and comparing diatom composition, Si limitation of diatom production was found to be more possible. The possibility of Si limitation was from 37% in 1980s to 50% in 1990s. Jiaozhou Bay ecosystem is becoming serious eutrophication, with notable increase of NO2-N, NO3-N and NH4-N from 0.1417 micromol/L, 0.5414 micromol/L, 1.7222 micromol/L in 1960s to 0.9551 micromol/L, 3.001 micromol/L, 8.0359 micromol/L in late 1990s respectively and prominent decrease of Si from 4.2614 micromol/L in 1980s to 1.5861 micromol/L in late 1990s; the nutrient structure is controlled by nitrogen; the main limiting nutrient is probably silicon; because of the Si limitation the phytoplankton community structure has changed drastically. PMID- 15900767 TI - Effect of Mo contents on properties of Mo/ZSM-5 zeolite catalyst for NOx reduction. AB - Mo/ZSM-5 catalysts with different Mo content were prepared by impregnation method. The effect of Mo content on the property of Mo/ZSM-5 catalysts and their performance for selective catalytic reduction(SCR) of NO with ammonia was investigated by XRD, ICP, XPS and NO-TPD respectively. The results showed that the catalytic activity of Mo/ZSM-5 for SCR of NO is strongly influenced by the Mo loading in HZSM-5 zeolites. The NOx conversion reached the highest value of 64.2% at 375 degrees C when Mo content is about 10.9%, and the temperatures at which the maximum of NOx conversion obtained were declined with the increase of Mo content. From XRD results, it can be seen that it exhibits the distinct interaction between Mo and HZSM-5 when Mo content is about 10.9% . This may result in a suitable phase structure in Mo/ZSM-5 catalyst, which is advantageous for NO reduction. XPS and NO-TPD results also showed that the catalytic activity of Mo/ZSM-5 may be related to the Mo percent on the surface. PMID- 15900768 TI - Variations of dominant microbial populations in groundwater in response to the leachate from Laogang Landfill. AB - Temporal changes of dominant microbial populations in groundwater in response to the leachate from Shanghai Laogang Landfill were investigated. Concentrations of dissolved redox-relevant species in groundwater suggested that the dominating redox process had changed from denitrification to methane-production/sulfate reduction due to landfilling. Dominant microbial populations were determined using restriction fragment length polymorphism( RFLP) analyses of 16S rRNA gene libraries, which were further studied by sequencing and phylogenetic analyses. The results indicated that obvious shifts of dominant microbial populations had occurred in groundwater in response to the pollution of leachate. The closest relatives of some dominant clones are accordant with the dominating redox processes determined by hydrochemical analyses, based on the GenBank's indications on the ability to perform redox reactions. PMID- 15900769 TI - Combined effect of US/PFS on the black liquor of making paper. AB - The reed pulping black liquor is used as a sample. Researches of (1) combined effect of ultrasound( US)/polyferric sulphate (PFS)/hydrogen peroxide (H2O2); (2) effects of the horn tip areas; (3) effect of dose of PFS were carried. Extrapolation of these experiments indicated treatment results are possibly related to complex chemical compositions in the reed pulping black liquor. PMID- 15900770 TI - Levels of polychlorinated biphenyls in 1,4-dichlorobenzene mothballs. AB - Although polychlorinated biphenyls (PCBs) are globally recognized pollutant, an understanding of their resources as by-products in chemical industry is poorly investigated. This paper presents data from a study that was conducted in order to determine the extent of PCBs in 1,4-dichlorobenzene( p-DCB) mothballs. Total PCBs and dioxin-like PCBs were detected in five mothball samples. Total PCB concentrations ranged from 328 ng/g to 1798 ng/g, while the levels of WHO-TEQ were between 0.16 pg/g and 13 pg/g. Mean concentrations of total PCBs and WHO-TEQ in the samples were 724 ng/g and 3. 2 pg/g, respectively. The highest level of PCB congeners was that of trichlorinated biphenyls (537 ng/g), which maybe due to the relatively low chlorination in the process of p-DCB production. A mechanism of the formation of PCBs from polychlorinated benzene in the presence of chlorine is also suggested. The results suggested that re-estimation on the risk of p-DCB products, especially the products for daily use such as mothballs, is expected. PMID- 15900771 TI - Leaching of metals on stabilization of metal sludge using cement based materials. AB - Toxicity characteristic leaching procedure(TCLP) of zinc plating sludge was carried out to assess the leaching potential of the sludge and the leachates were analyzed for heavy metals. The concentration of zinc, chromium, and lead in the leachate were 371.5 mg/L, 1.95 mg/L and 1.99 mg/L respectively. Solidification of zinc sludge was carried out using four different binder systems consisting of cement mortar, fly ash, clay and lime and cured for 28 d. The ratio of sludge added varied from 60% to 80% by volume. The solidified products were tested for metal fixing efficiency and physical strength. It was observed that the volume of sludge added that resulted in maximum metal stabilization was 60% for all the combinations, above which the metal fixation efficiency decreased resulting in high values of zinc in the leachate. Addition of 5% sodium silicate enhanced the chemical fixation of metals in all the binder systems. Among the four fixing agents studied, mixture of fly ash: lime, and cement mortar: lime stabilized zinc and other metals in the sludge effectively than other combinations. Addition of lime increased the stabilization of zinc whereas cement mortar increased the strength of the solidified product. PMID- 15900772 TI - Natural organic matter (NOM) removal from surface water by coagulation. AB - Poly-aluminum-chloride-sulfate(PACS) with different SO4(2-)/Al3+ mole ratios and bacicity(y) of 2.0 was synthesized using AlCl3 x 6H2O, Al2 (SO4 )3 x 18H2O and Na2CO3 as raw materials. The effect of SO4(2-)/Al3+ ratio on the performance of PACS for removal of natural organic matter(NOM) with humic-rich actual water was examined. It was found that PACS with SO4(2-)/Al3+ mole ratio of 0.0664 achieved the best NOM removal results and was selected to investigate its performance in comparison with PAC, FeCl3 and alum (Al2 (SO4)3 x 18H2O). The experimental results showed that the optimum NOM removals were achieved at pH 5.0-8.2 and the dose of about 5.0 mg/L as Al both for the selected PACS and PAC, at pH 5.0-6.0 and the dose of about 7.0 mg/L as Fe for FeCl3, and at pH 5.0-7.0 and the dose of about 7.0 mg/L as Al for alum, respectively. At the optimum conditions, the selected PACS achieved the best NOM removal result, followed by PAC, FeCl3, and then alum. The concentration of residual aluminum in treated water by the selected PACS and PAC under the optimum coagulant conditions was approximately 115 microg/L, which can completely comply with the regulated limits. PMID- 15900773 TI - Streptomyces avermitilis from marine. AB - The insecticidal strain 173 was isolated from marine source and its activity was explored by the bioassay of brine shrimp and Helicoverpa armigera. Based on morphological, physiological and molecular properties, the insecticidal strain 173 was identified as Streptomycetes avermitilis, which is the best insecticidal microorganism found in the terrestrial environment. The taxonomy of the strain 173, insecticidal spectrum and properties of the corresponding insecticidal antibiotics are reported. PMID- 15900774 TI - Comparison of Pb and Cd adsorption to the surface coatings and surficial sediments collected in Xianghai Wetland. AB - Surface coatings and surficial sediments were obtained in four natural waters in Xianghai Wetland in China to study the role of surface coatings and surficial sediments in controlling the transporting and cycling of heavy metals in aquatic environments. Pb and Cd adsorption to the surface coatings and surficial sediments were measured under controlled laboratory conditions(mineral salts solution with defined speciation, ionic strength 0.05 mol/L, 25 degrees C and pH 6.0 for surface coatings; and 0.005 mol/L CaCl2 solution, 25 degrees C and pH 6.0 for surficial sediments). The Langmuir adsorption isotherm was applied to estimate equilibrium coefficients of Pb and Cd adsorption to the surface coatings and surficial sediments, and the component analyses of surface coatings and surficial sediments were also carried out. Correlation analyses between the maximum adsorption of Pb and Cd (adsorption max ) and the components in the surface coatings and surficial sediments suggested that there was a statistically significant trend for Pb and Cd adsorption( adsorption max ) to the surface coatings to increase with increasing in contents of Fe and Mn oxides in the surface coatings and surficial sediments. And the metal adsorption abilities of surface coatings were much stronger than those of surficial sediments, highlighting that in the same water, i.e. at the same pH and initial metal concentrations, the metals (such as lead and cadmium) in supernatant were feasible to be adsorbed by surface coatings than surficial sediments. The more importance of surface coatings than surficial sediments for adsorbing and cycling of heavy metals in aquatic environments was evidenced. PMID- 15900775 TI - Changes in soil microbial community structure associated with two types of genetically engineered plants analyzing by PLFA. AB - With the rapid expansion of GEPs(genetically engineered plants), people are more and more concerned about the ecological risks brought by their release. Assessing the effect of GEPs on soil microbial ecology is indispensable to study their ecological risks. In our study, the phospholipids fatty acid ( PLFA) method was used to analyze the microbial community of soil samples collected from fields with two types of GEPs-Bt transgenic corn and PVY ( potato virus Y) cell protein gene transgenic potato. The principal components analysis (PCA) showed all controls were on the right of related GEPs samples along the PC1 (the first principal component) axis, which means a decrease of fungi in soils with genetically engineered crop since most of PLFAs that are strongly positively correlated with PC1 represent fungi. For samples collected from Bt transgenic cornfield, the ratios of gram-positive to gram-negative bacteria were less than those of controls. For samples of transgenic potato field, these ratios were lower than those of controls when soils were collected from deep layer (20-40 cm), but were higher when soils collected from surface layer(0-20 cm). For soils collected from 0-20 cm, the ratios of fungi to bacteria for all GEPs samples were at the same level. So were such rations for all controls. Changes of soil microbial community in two types of GEPs fields were detected in our study, but the causes and more information still needs further study. PMID- 15900776 TI - Subjective annoyance caused by indoor low-level and low frequency noise and control method. AB - The influence of low-level noise has not been widely noticed. This paper discovered that low-level and low frequency noise (A-weighted equivalent level Leq < 45 dB) causes higher probability of subjective annoyance. The fuzzy mathematic principle was applied to deal with the threshold level of subjective annoyance from noise in this study; there is preferable relationship between the indoor noise and noise annoyance at low frequency noise level. Study indicated at the same centered noise level, the change of annoyance probability is mainly caused by the change of the frequency spectrum characteristic of the indoor noise. Under low noise level environment, without change of the medium-low frequency noise, the slight increase of medium-high frequency noise level with the help of noise sheltering effect can significantly reduce the noise annoyance. This discovery brings a new resolution on how to improve the environmental quality of working or living places. A noise control model is given in this study according to the acoustic analysis. PMID- 15900777 TI - Investigation of polycyclic aromatic hydrocarbons from coal gasification. AB - The hazardous organic pollutants generated from coal gasification, such as polycyclic aromatic hydrocarbons(PAHs), are highly mutagenic and carcinogenic. More researchers have paid particular attention to them. Using air and steam as gasification medium, the experiments of three kinds of coals were carried out in a bench-scale atmospheric fluidized bed gasifier. The contents of the 16 PAHs specified by US EPA in raw coal, slag, bag house coke, cyclone coke and gas were measured by HPLC to study the contents of PAHs in raw coal and the effects of the inherent characters of coals on the formation and release of PAHs in coal gasification. The experimental results showed that the distributions of PAHs in the gasified products are similar to raw coals and the total-PAHs content in coal gasification is higher than in raw coal(except Coal C). The total-PAHs contents increase and then decrease with the rise of fixed carbon and sulfur of coal while there has an opposite variation when volatile matters content increase. The quantities of PAHs reduce with the increase of ash content or the drop of heating value during coal gasification. PMID- 15900778 TI - Structural and photocatalytic properties of TiO2 films fabricated on silicon substrates by MOCVD method. AB - Silicon ( 111) and Silicon(100) were employed for fabrication of TiO2 films by metal organic chemical vapor deposition( MOCVD). Titanium(IV) isopropoxide(Ti[O(C3H7)4] ) was used as a precursor. The as-deposited TiO2 films were characterized with FE-SEM, XRD and AFM. The photocatalytic properties were investigated by decomposition of aqueous Orange II . And UV-VIS photospectrometer was used for checking the absorption characteristics and photocatalytic degradation activity. The crystalline and structural properties of TiO2 film had crucial influences on the photodegradation efficiency. For MOCVD in-situ deposited films on Si substrates, the photoactivities varied following a shape of "M": at lower(350 degrees C ), middle(500 degrees C ) and higher(800 degrees C ) temperature of deposition, relative lower photodegradation activities were observed. At 400 degrees C and 700 degrees C of deposition, relative higher efficiencies of degradation were obtained, because one predominant crystallite orientation could be obtained as deposition at the temperature of two levels, especially a single anatase crystalline TiO2 film could be obtained at 700 degrees C. PMID- 15900779 TI - Biological formation of 5-aminolevulinic acid by photosynthetic bacteria. AB - In this study, 7 stains of Rhodopseudomonas sp. were selected from 36 photosynthetic bacteria stains storied in our laboratory. Rhodopseudomonas sp. strain 99-28 has the highest 5-aminolevulinic acid(ALA) production ability in these 7 strains. Rhodopseudomonas sp. 99-28 strain was mutated using ultraviolet radiation and a mutant strain L-1, which ALA production is higher than wild strain 99-28 about one times, was obtained. The elements affecting ALA formation of strain 99-28 and L-1 were studied. Under the optimal condition( pH 7.5, supplement of ALA dehydratase(ALAD) inhibitor, levulinic acid(LA) and precursors of ALA synthesis, glycine and succinat, 3000 Ix of light density), ALA formation of mutant L-1 was up to 22.15 mg/L. Strain L-1 was used to treat wastewater to remove COD(Cr) and produce ALA. ALA production was 2.819 mg/L, 1.531 mg/L, 2.166 mg/L, and 2.424 mg/L in monosodium glutamate wastewater(MGW), succotash wastewater(SW), brewage wastewater(BW), and citric acid wastewater(CAW) respectively. More than 90% of COD(Cr) was removed in four kinds of wastewater. When LA, glycin and succinate were supplied, ALA production was dramatically increased, however, COD(Cr) could hardly be removed. PMID- 15900780 TI - Determination of arsenic in air particulates and diesel exhaust particulates by spectrophotometry. AB - A method was developed for the determination of trace arsenic by spectrophotometry. The proposed method is rapid, simple, and inexpensive. This method can be used for sensitive determination of trace arsenic in environmental samples and especially in air particulates. The results obtained by this method as a proposed method were compared with those obtained by hydride generation atomic absorption spectrometry as a popular reported method for the determination of arsenic and an excellent agreement was found between them. The method was also used for determination of arsenic associated with airborne particulate matter and diesel exhaust particulates. The results showed that considerable amount of arsenic are associated with diesel engine particulates. The variation in concentration of arsenic was also investigated. The atmospheric concentration of arsenic was different in different sampling stations was dependent to the traffic density. PMID- 15900781 TI - Adsorption of direct green B on mixed hydroxy-Fe-Al pillared montmorillonite with large basal spacing. AB - Through cation-exchanging of montmorillonite, mixed hydroxy Fe-Al-pillared montmorillonites with large basal spacing were synthesized. The application of these mixed hydroxy Fe-Al-pillared montmorillonites as adsorbents in the areas of environmental protection and pollution control was investigated. Adsorption of Direct Green B, an organic dye, on OH--Fe--, OH--Al--, OH--Fe--Al-- pillared montmorillonites was studied. Results showed that these pillared montmorillonites had much better adsorption properties than traditional Na-organo-montmorillonite. It was also found that pH of the solutions had great effect on the performance of these hydroxy-metal pillared montmorillonites in treating the wastewater containing Direct Green B. PMID- 15900782 TI - Chemical and biological flocculation process to treat municipal sewage and analysis of biological function. AB - The pilot-scale experimental apparatus and the procedure of the chemical and biological flocculation process to verify the feasibility in treating Shanghai municipal sewage were introduced in this paper. In addition, the biological function of the process was discussed. The results of optimal running showed that in the reaction tank, the concentration of mixed liquor suspended solid(MLSS) was 2 g/L, hydraulic retention time(HRT) was 35 min, dosage of liquid polyaluminium chloride(PAC) was 60 mg/L, and the concentration of polyacrylamide(PAM) was 0.5 mg/L. The effluent average concentrations of COD(Cr), TP, SS and BOD5 were 50 mg/L, 0.62 mg/L, 18 mg/L, and 17 mg/L, respectively. These were better than the designed demand. In addition, the existence of biological degradation in this system was proven by several methods. The removal efficiencies of the chemical and biological flocculation process were 20% higher than that of the chemical flocculation process above at the same coagulant dosage. The treatment process under different situations was evaluated on a pilot-scale experiment, and the results provided magnificent parameters and optimal condition for future operation of the plant. PMID- 15900783 TI - Heavy metal concentrations in redeveloping soil of mine spoil under plantations of certain native woody species in dry tropical environment, India. AB - Total concentration of heavy metals (Cd, Cr, Cu, Fe, Pb, Ni, Mn and Zn) was estimated in the redeveloping soil of mine spoil under 5-yr old plantations of four woody species namely: Albizia lebbeck, Albizia procera, Tectona grandis and Dendrocalamus strictus. The data recorded in the present study were compared with other unplanted coal mine spoil colliery, which was around to the study site and adjoining area of dry tropical forest. Among all the heavy metals, the maximum concentration was found for Fe and minimum for Cd. However, among all four species, total concentrations of these heavy metals were recorded maximally in the plantation plots of T. grandis except for Fe, while minimally in A. lebbeck except for Zn, whereas, the maximum concentration of Fe and Zn was in the plantation plots of D. strictus and A. procera. Statistical analysis revealed significant differences due to species for all the heavy metals except Cu. Among four species, A. lebbeck, A. procera and D. strictus showed more efficient for reducing heavy metal concentrations whereas T. grandis was not more effective to reduce heavy metal concentrations in redeveloping soil of mine spoil. PMID- 15900784 TI - Comment on "removal of heavy metals from aqueous solution by carbon nanotubes: adsorption equilibrium and kinetics" by Li, Y.H., Di, Z.C., Luan, Z. K., Ding, J., Zuo, H., Wu, X. Q., Xu, C.L. And Wu, D.H. PMID- 15900785 TI - Act 422 reduces recoupment time. PMID- 15900786 TI - Report of the Arkansas Health Care Access Foundation Inc. PMID- 15900787 TI - Passing the torch. Retiring AMS executive leaves a legacy of leadership. PMID- 15900789 TI - Arkansas Department of Health 2004 report. PMID- 15900792 TI - Health information technology: from theory to practice. AFMC helps Arkansas physicians rise to the challenge. PMID- 15900793 TI - Overview of biologic agents in medicine and dermatology. AB - Three agents have recently been approved by the Food and Drug Administration for the treatment of chronic plaque psoriasis: alefacept, efalizumab, and etanercept. The field of dermatology has now entered a new era, joining other disciplines of medicine that have been using biologic agents for decades. These new therapies offer psoriatic patients the potential for safe and effective long-term management of this disease. This article reviews how an increased understanding of the pathophysiology of psoriasis led to the development of these products. PMID- 15900794 TI - Alefacept for the treatment of psoriasis: a review of the current literature and practical suggestions for everyday clinical use. AB - Alefacept is the first biologic therapy approved by the United States Food and Drug Administration for the management of patients with moderate-to-severe chronic plaque psoriasis. Alefacept is a fully human fusion protein with a dual mechanism of action inhibiting T-cell activation as well as selectively reducing memory T cells. More than 2000 psoriasis patients have been treated with alefacept in clinical trials. These studies reveal an excellent clinical response, with 33% of patients achieving a Psoriasis Area Severity Index (PASI) score of 75 and 57% of patients achieving a PASI 50 after one course of alefacept. Patients achieving both PASI 75 and PASI 50 have significant improvements in their quality of life. The best responders can have long remissions, and there is a tendency toward continued improvement with subsequent courses of alefacept. The safety profile over the short and intermediate term is excellent. Preliminary data regarding alternate dosing regimens, transitioning patients from conventional systemics to alefacept, and combining alefacept with ultraviolet light therapy will be highlighted. We also will discuss our practical approach to patient selection, CD4 monitoring, management of infections while on alefacept, as well as decisions regarding retreatment and combination therapy. PMID- 15900795 TI - Clinical considerations of efalizumab therapy in patients with psoriasis. AB - Psoriasis is a chronic, incurable disease that often requires decades of therapy to maintain disease control. Efalizumab is a recombinant monoclonal IgG1 antibody approved for use in patients with chronic moderate-to-severe plaque psoriasis. To date, efalizumab has been evaluated extensively in more than 3500 patients, including in studies that have evaluated its efficacy and safety during extended use. Just as psoriasis fluctuates in severity, the response to treatment with efalizumab can vary among patients. On the basis of my personal experience managing patients in and out of clinical trials, most patients benefit from efalizumab. The possibility exists of intercurrent events during efalizumab therapy, such as the development of a transient localized papular eruption or mild arthralgia or, in a few patients, a generalized inflammatory flare or severe arthralgia. However, there are techniques to potentially manage these events in a manner that maximizes patient comfort and compliance. If dermatologists become comfortable recognizing the subset of patients who are overall excellent responders but develop a papular eruption or mild and manageable arthralgia, they will be able to readily incorporate this effective biologic into their daily practice. In this article, clinical trial data and case reports illustrate recommended patient management techniques and the substantial long-term benefits that psoriasis patients may realize with efalizumab therapy. PMID- 15900796 TI - The treatment of psoriasis with etanercept. AB - Etanercept is a tumor necrosis factor alpha (TNF-alpha) inhibitor approved for the treatment of psoriasis. Etanercept is a soluble version of the tumor necrosis factor receptor (TNFR) that neutralizes the proinflammatory activity of TNF alpha, a molecule central to the pathogenesis of psoriasis. Patients receiving etanercept continuously during both 12 and 24 weeks show a significant reduction in the signs of psoriasis. Further, higher doses of etanercept provide better efficacy. Both clinical trial and postmarketing experience with etanercept is extensive and, thus, etanercept has a well-defined safety and tolerability profile. With appropriate patient selection and follow-up, etanercept therapy has a very good benefit-to-risk ratio and represents a convenient option for patients with moderate-to-severe psoriasis. PMID- 15900797 TI - Combining traditional agents and biologics for the treatment of psoriasis. AB - Psoriasis patients deserve long-term control of their disease with optimal safety. Traditional agents (methotrexate, cyclosporine, retinoids, and photochemotherapy [PUVS]), although providing excellent short-term control, may produce acute or chronic toxicities, thus limiting their usage. Dermatologists are well versed in combination and rotational therapies for psoriasis, using these and other agents. With the advent of biologic therapies (three currently approved, and others pending), the potential for safer long-term psoriasis control is being realized. A review of the literature, plus our personal experience in using combinations of traditional agents and biologics, is presented. PMID- 15900798 TI - Psoriatic arthritis and psoriasis: need for a multidisciplinary approach. AB - Psoriatic arthritis is a progressive and often destructive form of seronegative inflammatory arthritis that is associated with psoriasis. It can be difficult to diagnose because it can present in a number of different ways, often indistinguishable from noninflammatory arthropathies such as osteoarthritis as well as inflammatory arthritis such as gout. Severe arthritis may be observed in the absence of psoriasis, or mild arthritis may be seen in the presence of moderate-to-severe psoriasis. A high index of suspicion, screening of psoriasis patients, and close follow-up and evaluation with rheumatology often is needed to make the diagnosis. Early recognition of the disorder and timely therapy can prevent long-term complications, such as permanent joint destruction and disability. With the advent of biologic agents, we are better equipped to manage psoriatic arthritis today. Because dermatologists are on the front-line of psoriasis management, we are perfectly poised to identify and help improve care for patients who suffer from both psoriasis and psoriatic arthritis. PMID- 15900799 TI - The impact of biologics on the quality of life of psoriasis patients and the economics of psoriasis care. AB - Psoriasis has a tremendous impact on patients' lives, affecting them physically, psychologically, and socially. Thus, it is not merely a cosmetic concern and often warrants appropriately aggressive treatment. Traditional treatments for moderate-to-severe psoriasis include phototherapy, oral retinoids, methotrexate, and cyclosporine. Newer biologics combat the immunologic mechanism responsible for psoriasis and, to date, carry a more favorable side effect profile. We examined the impact on quality of life of biologics and assessed their total direct costs to psoriasis patients. Biologic treatments significantly improve the quality of life of psoriasis patients; however, they cost significantly more than traditional therapies. This difference calls for physicians to weigh the costs and benefits of biologic therapies and compare them to those of traditional treatments when considering care for psoriasis patients. PMID- 15900800 TI - Potential future therapies for psoriasis. PMID- 15900801 TI - [Child and adolescent psychosomatic medicine]. PMID- 15900802 TI - [Relevance and results of Theory of Mind research for autism and other psychiatric disorders]. AB - OBJECTIVES: Disorders of the autistic spectrum are basically characterised by a triad of symptoms: dysfunction of social interaction, communication deficits, and stereotyped behaviour patterns and interests. One of the most prominent approaches to explaining these abnormalities is the "Theory of Mind" (Baron-Cohen et al., 1885). METHODS: The present review discusses and critically examines the ongoing research and recapitulates the essential findings of the last ten years, focussing on their methodological quality and utility to explain other psychiatric disorders. RESULTS: Despite considerable research efforts in this field, the existing concepts do not constitute a consistent framework for analysing the development of autism spectrum disorders. CONCLUSIONS: Future research should aim to verify the impact of the existing theoretical models and to emphasise the similarities of the different concepts in order to gain specific information about potential causal factors of autism. PMID- 15900803 TI - [Outpatient treatment of childhood and adolescent obesity]. AB - OBJECTIVES: Obesity in childhood is an increasing problem in both industrial and developing countries. Prevalence rates are up 15% in the U.S.A. and up 10% in Europe. The present paper gives an overview of current research on medical and psychosocial problems associated with childhood obesity. Treatment options are introduced and 24 studies are reviewed in detail with regard to treatment characteristics and outcome. METHODS: Medline and PsycINFO searches for outpatient treatment studies on childhood obesity between 1966 and 2003 were carried out. RESULTS: The limited comparability of the studies necessarily restricts their generalizability. CONCLUSIONS: Implications for the treatment of childhood obesity and further research are discussed. PMID- 15900804 TI - [Aggressive behaviour and substance abuse among schizophrenic adolescents compared to antisocial adolescents--a follow-up study]. AB - OBJECTIVES: The objective of this study was to analyze aggressive behaviour towards others by schizophrenic as opposed to antisocial adolescents, and the influence of substance abuse before, during and after their hospitalization. METHODS: We analyzed 21 schizophrenic adolescents and compared their aggressive behaviour and their substance abuse to that of 21 antisocial juveniles before and during their hospitalization and again at the time of a follow-up interview. The two samples were matched for age, sex and intelligence. In a first step, data were gathered from the hospital records, in a second step, for follow-up data we conducted standardized telephone interview with the patient and his or her parent or caregiver. Within the analysis we focused on aggressive behaviour towards other people and objects, as well as on criminal acts and regular substance abuse. RESULTS: We found less aggressive behaviour among psychotic patients during and post-hospitalization than among their antisocial counterparts. As inpatients, the acutely psychotic juveniles were at higher risk for aggressive acts, but adequate treatment subdued their offensive behaviour. In the long term, there were fewer criminal arrests among psychotic patients. Only in connection with their substance abuse, their aggressive misconduct towards others increased. CONCLUSIONS: Our results suggest that drug treatment during adolescence might help to lessen the risk of aggressive behaviour towards others. PMID- 15900805 TI - [Evaluation with the "Psychotherapie Basisdokumentation" for Children and Adolescents: Psy-BaDo-KJ--a questionnaire for quality assurance and evaluation of psychotherapy for children and adolescents]. AB - OBJECTIVES: Currently only few instruments exist to evaluate psychotherapy for children and adolescents (Mattejat & Remschmidt, 1993). This study presents the development of a new instrument: the "Psychotherapie Basisdokumentation fur Kinder und Jugendliche: Psy-BaDo-KJ" (Winter, 2002). This instrument assesses individual therapy goals and the outcome of therapy on the basis of information from children and adolescents, parents and therapists. METHODS: We based our instrument on a questionnaire originally developed for adults: the "Psychotherapie Basisdokumentation" (Psy-BaDo, Heuft & Senf, 1998). To develop the new instrument, we conducted expert interviews that were then analysed qualitatively. Between January 2002 and June 2003 all inpatients (> or = 14 years) as well as their parents and therapists were given questionnaires. To date, data for 35 patients have been evaluated and included in the current study. The diagnoses were recorded according to ICD-10 (Remschmidt et al., 2002). RESULTS: The study shows that patients, parents and therapists usually formulate two or three individual therapy goals but with different content. While the patients mention primarily goals focussed on somatic health, the focus of parents and therapists is predominantly on psychological and mental health. According to the outcome of therapy, the level of agreement among patients, parents and therapists is high. Patients tend to be more optimistic than their parents and therapists. CONCLUSIONS: The "Psychotherapie Basisdokumentation fur Kinder und Jugendliche" (Psy-BaDo-KJ) is a new instrument and can be used efficiently for quality assurance and evaluation of psychotherapy for children and adolescents. PMID- 15900806 TI - [Comparing assessment methods of infant emotionality]. AB - OBJECTIVES: The concept of infant emotionality (temperament) is frequently used in the fields of developmental psychopathology, developmental psychology, and child and adolescent psychiatry. However, a valid assessment of the construct has to deal with some difficulties. Parent reports and behaviour observations of the interaction between caregiver and infant may be biased by parental characteristics, while laboratory assessment procedures often have not been sufficiently validated. METHODS: In the present study, three dimensions of temperament were assessed at three ages during the first year of life using three different measurement approaches. Convergent validity, discriminant validity and the associations of the temperament measures with maternal characteristics, i.e. depression, anxiety and educational status were analyzed. The study group consisted of 101 healthy first-born infants and their primary caregivers. At the ages of 4, 8 and 12 months, positive emotionality, negative emotionality and withdrawal/anxiety were assessed by means of a parent questionnaire, naturalistic behavioural observations of the caregiver-infant interaction, and by means of laboratory routines. RESULTS: Aside from just two exceptions, there were significant convergent correlations between the different measures at each age. Particularly the laboratory routines revealed a good degree of discriminant validity. Questionnaire scores often correlated with the mothers' characteristics. These correlations were independent of the associations with the observational data. CONCLUSIONS: Therewith, parent reports include an objective as well as a subjective component. PMID- 15900807 TI - A review of longitudinal studies on youth predictors of adulthood physical activity. AB - The aim of this article was to review the literature on the youth predictors of physical activity in adulthood. The outcome measure in the reviewed studies was the level and types of physical activity at adult age. The youth predictors here included different measures of physical activity, health and fitness as well as some social and behavioral variables measured in childhood or in adolescence. The goal was to identify the elements in youth that are associated with a high level of physical activity in adulthood. This information can be used by health promoters who plan physical activity programs for young people to enhance lifelong physical activity. This literature review is also addressed to researchers to provide ideas for further studies in this area. PMID- 15900808 TI - Physical activity in adolescence. A review with clinical suggestions. AB - Despite some inconsistencies in research methodologies, most findings support a positive correlation between participation in physical activities and well-being in adulthood. The results are consistent across the life span of both genders. Favorable connection between physical exercise to physical, psychological, emotional and educational benefits has been constantly proven. Despite such results a comparison between present to past findings show a global tendency for sedentary life style and reduced physical activities in many countries across ages and genders. There are claims that achieving an adult healthy life style is rooted in habits acquired at early ages, thus pointing at childhood and adolescence as the starting point of an active and healthy adulthood. The present article reviews the current literature and findings relating to physical activity with better health and an emphasis on adolescence. Factors correlated to participation of adolescents in physical activities are presented and some clinical issues to promote such activity are discussed. The authors strongly recommend enhanced initiation of community based easily accessed physical exercise programs, for children and adolescents. PMID- 15900809 TI - A review of physical activity and well-being. AB - The World Health Organization defines health as a state of complete physical, mental and social well-being. Based on this declaration the construct of well being has been researched. Many researchers have set a clear path between physical exercise and feeling better or the connection of reduced physical activity and diminished health. Nevertheless the allusive subjective psychological construct of well-being has not been directly connected to physical activity. Despite abundance of technical evidence that supports notions of correlations between physical exercise and well-being, the scientific proof is not within our reach yet. Some of the basic reasons are the facts that the definition of well-being is unclear, not many RCT's (Randomized Control Trials) have been performed, dose related results are scarce and many articles use small populations and different methodology. Until an actual research based connection will be found between physical exercise and well-being, the authors strongly recommend physical activity as part of everyone's leisure time, since there are enough proven physical mental, and social benefits to physical activity besides well-being. PMID- 15900810 TI - Adapted physical activities for the intellectually challenged adolescent: psychomotor characteristics and implications for programming and motor intervention. AB - Intellectually challenged adolescents with varying degrees of functioning share common behavioral and psychomotor characteristics. A specially designed instructional approach and positive social attitude are necessary when dealing with this population. Research reveals that many of these individuals have developmental delays in the acquisition of basic motor skills. When compared as a group to their non-handicapped peers, intellectually challenged adolescents display low physical fitness and have perceptual-motor difficulties, which affect their learning. The major difficulty these individuals are faced with stems from language comprehension barriers and their failure to grasp instructions. In addition, some possess physical characteristics, which pose constraints in learning and performing of motor skills. A social attitude of equality and acceptance plays a major role in their successful inclusion in society. Researchers agree that the limitations mentioned above, affect the motivation of these individuals and lead to a lack of opportunity for regular participation in movement, physical activities and sports. This situation leads to low performance in the motor domain and in other areas as well. This article describes the behavioral and psychomotor characteristics of intellectually challenged individuals and offers general and specific recommendations for instruction and intervention. PMID- 15900811 TI - Physical inactivity and socioeconomic status in Canadian adolescents. AB - Physical activity is an important determinant of population health. In this paper, we examine levels of physical activity among Canadian adolescents in relation to socioeconomic status. METHODS: Data from the Canadian Community Health Survey, Cycle 2.1, conducted in 2003, were used to evaluate the association between socioeconomic status and physical activity. This survey included 9,294 males between 12 and 20 years of age, and 9,147 females in the same age range. RESULTS: Among adolescents aged 12 to 20 years, 55.9% of males and 39.2% of females were physically active. The proportion of subjects in the active category tended to decrease with age in both males and females. Adolescents from low income families were more physically inactive (odds ratio=1.30, 95% confidence interval=1.29-1.32) than those from high income families. CONCLUSION: Physical inactivity in adolescence represents an important population health issue in Canada. Physical activity levels tended to be higher among males, younger people, and those with higher socioeconomic status. Multi sectoral policies and initiatives are needed to create environments that promote physical activity among Canadian youth. PMID- 15900812 TI - Biochemical evaluation of antioxidant function after a controlled optimum physical exercise among adolescents. AB - Sensible physical exercise is shown to prevent certain neurovascular problems. However, in recent times, non-traumatic sudden death in young athletes has been observed and the incidence level is always very disturbing, because of the spontaneous nature of the occurrence. It most commonly occurs fivefold more in male than female athletes. Although it is believed that congenital cardiovascular disease is the leading cause of non-traumatic sudden athletic death, however, sudden physical alteration in the biochemical composition of the body system may, at least in part, play an important role. The role of antioxidants in the general maintenance of homeostasis has already been established. In this study, total antioxidant function in athletes subjected to controlled physical exercise was evaluated to determine the extent to which intensive physical exercise could alter the health conditions if adequate actions are not taken to adjust the biochemistry of the body system. Ten male field-track athletes were exercised using a fixed workload treadmill test. Blood samples were drawn before and after the exercise. The subjects exercised almost to their maximum running distance at a higher "fatigue" workload for a maximum of 20 min each day for five days. The lymphocytes' total antioxidant function was measured by addition of a peroxide (oxidative stress) to complete the medium. Lymphocyte growth response with peroxide was reported as a percentile of growth responses from a reference range of apparently healthy individuals. Values below the 25th percentile indicate a deficient antioxidant function. The results showed that although vitamin B3, B6, and B12 requirements were normal, there was a consistent low value in the total percentile of vitamins B1 (<79%), B2 (<54%), folate (<33%, and biotin (<70%). There was a dramatic decrease in the mean values of antioxidant function (38.1%) in all the subjects as opposed to the reference range of <75%. Overall reduction antioxidant function indicates decreased ability to resist oxidative stress, or an increased oxidant load, suggesting increased antioxidant utilization and/or cellular "tear and wear" scenario. It is therefore suggested that after intensive physical exercise, antioxidant functions should be monitored and supplemented whenever necessary to maintain the integrity of the cellular function. PMID- 15900813 TI - Don't stop, don't stop: physical activity and adolescence. AB - The aim was to assess whether physical activity decreases during adolescence, whether this decrease depends on the gender, whether physical activity is related to personal, family, and school factors, and whether it is associated to healthy behaviors in a sample of adolescents. Data were drawn from a survey carried out in 2001 among in-school Catalan adolescents aged 14-19 years. Subjects were divided in two groups: physical activity (N=4,185, 43.5% females) and no-physical activity (N=2,743; 68.9% females). Personal, family, school and lifestyles' variables were compared. Chi-square and Odds Ratio were used to compare qualitative variables and Student's t to compare quantitative variables. For the multivariate analysis, all statistically significant variables in the univariate analysis in each of the four groups of variables (plus age) were introduced in a non-conditioned multiple regression. Analysis was performed separately by gender. Physical activity was significantly more frequent among males and decreased with age. Globally, physically active youth perceived themselves as healthier and happier with their body image, they showed a better relationship with their parents, were better connected to school, and exhibited healthier lifestyles. As physical activity has important benefits on health, health professionals dealing with adolescents should encourage adolescents to keep practicing. This message must be specially directed to females. PMID- 15900814 TI - Prevalence of obesity and lack of physical activity among Kentucky adolescents. AB - Lack of physical activity and overweight status continues to be a significant health problem in the United States. To assess the actual prevalence of these problems, we reviewed data from the School-based Health Promotion Centers in one middle school, and one High school in central Kentucky. A total of 232 6th graders and 607 9th graders were included. A total of 92% of 6th graders and 45% of 9th graders reported complete lack of regular exercise. 37% of 9th graders and 59% of 6th graders had inappropriate nutrition, 47% of 9th graders and 33% of 6th graders had Body Mass Index over the 85th percentile for age. Among overweight adolescents, only 16% of 9th graders and 1% of 6th graders thought they were overweight. Since the body mass index data is derived from actual objective measurement, we conclude that overweight status is a significant problem in Kentucky and that the actual numbers may be higher than those reported for the State based on surveys and estimates. Furthermore, many adolescents are not aware of their weight problem and most of them do not participate in regular exercise. PMID- 15900815 TI - Hydrotherapy combined with Snoezelen multi-sensory therapy. AB - The aim of this article is to present a new and challenging model of treatment that combines two therapeutic interventions: hydrotherapy and Snoezelen or controlled multisensory stimulation. The combination of the two therapeutic approaches enhances the treatment effect by utilizing the unique characteristics of each approach. We believe that this combined model will further enhance each media to the benefit of the clients and create a new intervention approach. This article relates to a hydrotherapy swimming pool facility that has been established at the Williams Island Therapeutic Swimming and Recreation Center, Beit Issie Shapiro, Raanana in Israel, after acquiring many years of experience and gaining substantial knowledge both in the field of hydrotherapy and Snoezelen intervention. Beit Issie Shapiro is a non-profit community organization providing a range of services for children with developmental disabilities and their families. The organization provides direct services for nearly 6,000 children and adults each year. This article provides an overview of hydrotherapy and Snoezelen and presents a case study, which will demonstrate the new model of treatment and show how this new and innovative form of therapy can be used as a successful intervention. We believe it will open a path to enriching the repertoire of therapists helping people with special needs. This article is also addressed to researchers to provide ideas for further studies in this area. PMID- 15900816 TI - Physical fitness and adolescence. AB - Children, adolescents and adults all benefit from physical exercise and physical fitness can also bring well-being to the person. Tracking physical fitness and activity over time from childhood, adolescence into adulthood will provide information on the benefits and also increase over knowledge on intervention and intervention programs. Many children and adolescents participate in physical activity, but this activity decline in adulthood. This short review provide information from several studies on the benefit of physical exercise and fitness through childhood, adolescence into early adulthood. PMID- 15900817 TI - Neonatal teeth association with Down's syndrome. A case report. AB - A case of natal teeth in a four-day-old female brought by a forty four year old mother is presented. These natal teeth were present in the upper and lower jaws. Of special interest was the shape and positions of these especially the upper tooth in the left canine region and the fact that these teeth were present in a child with Down's syndrome features. The anxiety of the mother and the desire to get them removed immediately underlines the superstition and misconceptions with which natal teeth are viewed. The multidisciplinary approach to management is also highlighted. PMID- 15900818 TI - [Cost analysis of operating the oral health service of the regional hospital center of Ziguinchor (Senegal)]. AB - This study related to the analysis of the cost of operation of the service of oral health in the regional hospital complex of Ziguinchor for the year 2002. Its objective is to analyze the costs of operation of the service in order to allow a good covering of the costs while guaranteeing the accessibility to care and the perennity of the offer of service. The methodology used for the determination of the costs was the method of the complete costs (or method of the homogeneous sections). The various services provided by the service were identified. The resources mobilized for the realization of these services were determined and developed. They were classified in direct loads, i.e. directly ascribable to the provided service and in indirect loads when the resources are common to more than one service. The charge of the indirect loads was done thanks to a scale. The total cost of operation of the service of stomatology for year 2002 amounts to 11464943 F CFA. Two financial backers intervened with the financing of the service. The public funds support 75% of the loads and the community 25%. The cost to be recovered is 2845513 FCFA. The rate of covering is surplus for all the services safe for the oral surgery, the obturations with the composites and radiography. It is thus of primary importance to re-examine tariffing in force with the fall. This fall of tariffing will certainly make it possible to take restraint measures of the exemptions from payment. The covering of the costs by the means of the Community participation requires the installation of a tariffing likely to ensure the perennity of the services and to avoid exclusion. PMID- 15900819 TI - Need for preventive/interceptive orthodontic treatment among 7-10-year-old children in Ibadan, Nigeria: an epidemiological survey. AB - The aim of this epidemiological survey was to assess the level of needs for preventive/interceptive orthodontics among 7-10-year-old children in Ibadan, Nigeria, which is the second vigilance period in the study of developing occlusions. The sample consisted of 493 school children--237 (48.1%) boys and 256 (51.9%) girls randomly selected from various schools and different socio-economic groups. They were all examined in their school environments under natural illumination and the findings entered into a pre-structured form. The study revealed that about 27% of the children had need for one form of preventive/interceptive orthodontic treatment or the other with some having multiple needs. Some of the needs detected were as follows: prolonged retention of primary teeth (6.9%), cross bite (10.3%), early loss of primary teeth (4.3%), oral habits (7.3%), over jet of more than 6mm (4.9%), carious lesions (14.8%) crowding (18.9%) and clinically missing upper lateral incisors (3.7%). The relationship between observed occlusal discrepancies and oral habits was found very statistically significant (p=0.001). No gender differences were noted among the children with preventive/interceptive needs (p>0.05). Children from the working class accounted for 55.2% of the study sample, while 44.8% were from the middle class families. It was concluded that over one third of these children would benefit from preventive interceptive orthodontics and that such care should be encouraged in our environment due to their known numerous benefits especially in a country like Nigeria where many can not afford the cost of comprehensive orthodontic services. PMID- 15900820 TI - [Asepsis and antisepsis in dental offices in Dakar]. AB - To evaluate hygiene in dental offices in Dakar, the authors questioned 64 public, private and semi-private dentists. From 62 practitioners who completed their questionnaire, it appeared that the dry-heated sterilizer (poupinel) was 5 times more used than the autoclave; The almost practitioners sayed respecting the sterilisation process and 66% of them said wearing gloves, masks and glasses during interventions. In spite of the hope these results give, this study is limited because it doesn't take into account many aspects of asepsis and antisepsis in dental office which require a rigorous application next to a personal who needs to be better trained. PMID- 15900821 TI - Traditional oral health practices among Kanuri women of Borno State, Nigeria. AB - A structured questionnaire was administered on 495 women (urban 339, rural 156) from two LGAs of Borno State, Nigeria, using the interviewer method. The age range of the subjects was 12 to 80 years with a mean age (+/- SD) of 35.7 +/- 13.44 years. Majority (83.5%) did not have any formal education. Oral hygiene tools used by the respondents included toothbrush/paste 36 (7.9%), chewing stick 250 (54.9%), charcoal 159 (34.9%) and ordinary water 10 (2.2%). Of those using chewing sticks; 168 (67.2%) use Salvadora persica, 36 (14.4%) use Azadirachta indica and 46 (18.4%) use Eucalyptus camaldulensis. Forty (8.1%) of the respondents do not clean their teeth at all. Strong association was found between choice of teeth cleaning material and educational level (P=0.000). Three hundred and one (60.8%) of the respondents stain their teeth with flowers of Solanum incanum or Nicotania tabacum while, 218 (44.0%) perform tattooing of lip or gingivae and of this number 213 (97.7%) performed tattooing before marriage. Tattooing is usually performed without local anaesthesia with thorns of Balanites aegyptiaca and a mixture of charcoal & seeds of Acacia nilotica var. tomentosa as pigments. It is concluded that traditional oral health practices still constitute important part of the lifestyle in the study population. PMID- 15900822 TI - Orthodontic treatment need and demand in a group of Nigerian adults: a teaching hospital-based study. AB - Adults may be less influenced by peer perceptions and are, generally, more stable in their concerns about appearance compared to adolescents. This study aimed at assessing the level of objective orthodontic treatment need and demand for such care among the Nigerian adults in a referral centre (University College Hospital, Ibadan) as well as their social class. The Dental Aesthetic Index was used to assess the objective orthodontic treatment need while the social class of the subjects was according to the Registrar General's social class. During the study period (January 2001-September 2002), 49 adults aged 20-55 years with mean age of 25+ 6.3 (SD) years accounting for 27.5% of all orthodontic patients were seen. Equal proportions of the subjects had normal or minor malocclusions needing no treatment or slight treatment need and very severe or handicapping malocclusions with treatment considered mandatory (32.6% each). Next were severe malocclusions with treatment highly desirable accounting for 20.4%, followed by definite malocclusions with treatment elective (14.3%). Majority (71.4%) of the subjects was of the middle class families. PMID- 15900823 TI - [Oral health care practices in Abidjan]. AB - Abidjan, the economical capital of Ivory Coast has a CAO of 2.7 with index O practically nil. The professional density is one surgeon for 12.000 inhabitants. Abidjan population, despite the requirements in increasing care has not recourse to the dental surgeons for their assumption of responsibility. The general objective of our study is to describe the therapeutic itinerary in odonto stomatology of the Abidjan population. We have realised a questionnaire survey beside of population sample. The results show that 25 per cent of our population gives up care despite the presence of pain or embarrassment. Our study has allowed highlighting all the other recourses to the care. Indeed, the people who give up the care have much other recourse which has be identified in order of importance: the modern and traditional self-medication. Let us note in our study, that the choice of dental surgeon knows a constant evolution from the first to the third recourse. The lack of financial means constitutes however the first obstacle at the accessibility to the oral care in Ivory Coast. PMID- 15900824 TI - [African research in odontostomatology and international cooperation]. AB - In this study, the authors measure the efficacy of the African odonto stomatologic research in the International Assocation for Dental Research (I.A.D.R.) and value the impact of the international cooperation upon that research. This evaluation is realized from the number of publications presented by the African countries at the congressess of I.A.D.R. The obtained results show: 1. The African odonto-stomatologic is the less successful in the world and produced, in 1997, 0.4% of the studies published on the world, 2. Some of the African countries are able to produce scientific results of International valour (South Africa, Egypt and Nigeria in a less degree). Generally, these countries work jointly with anglo-saxon universities. 3. During these last two years, Cameroon obtained very encouraging results thanks to his collaboration with the University of Bordeaux 2 (France), represented by the regretted Professeur Ph. NDOBO-EPOY. PMID- 15900825 TI - [Pulmonary tuberculosis (in adults)]. PMID- 15900826 TI - [Role of computed tomography and magnetic resonance imaging in the diagnosis of hepatitis and liver cirrhosis]. AB - Computed tomography (CT) and Magnetic Resonance Imaging (MRI) have no significant impact in the evaluation of diffuse liver disease. Cirrhosis and hepatitis are not of specific imaging findings, the image of cirrhosis is depending on degree of disease. Nodular lesions are frequent findings in cirrhotic livers. For differentiation of regenerative nodules, dysplastic nodules and hepatocellular carcinoma CT and MRI are playing their role in localization and characterization of these lesions. Sensitivity and specificity are varying, depending on the technical applications of CT and MRI, and the application of contrast materials. MRI is superior in characterizing the lesions due to the different appearance of the lesions in different sequences. CT is superior as the staging modality. Complications of cirrhosis like ascites, varices of the oesophageal veins are diagnosed. CT and MRI are necessary when interventional or surgical procedures are planned or for therapy surveillance. PMID- 15900827 TI - [Value of ultrasound and doppler sonography in chronic hepatitis and liver cirrhosis]. AB - Ultrasound is unquestionably of great importance in the diagnosis of liver diseases due to the technical improvements of the last years. Indications for an ultrasound examination are the primary diagnosis of chronic liver disease and the screening for hepatocellular carcinoma. Apart from the diagnosis of portal hypertension in liver cirrhosis a portal vein thrombosis should be detected or excluded in cases of new or refractory ascites. The Budd-Chiari syndrome is another rare but important indication. There is a considerable improvement in the differential diagnosis of focal liver lesions especially due to contrast-enhanced sonography. Using phase-inversion harmonic sonography it is possible to detect tumor vascularity in a very sensitive manner. The accuracy of contrast-enhanced sonography is comparable with helical-CT or MRT, and contrast-enhanced sonography is very useful in the evaluation of response to ablation therapy of hepatocellular carcinoma. PMID- 15900828 TI - [Evaluation of non-invasive scores in the follow-up of chronic hepatitis]. AB - To differentiate biliary diseases the biopsy of liver tissue for histological examination is still the method of choice (golden standard). By histology the exact grading (inflammatory activity) and staging (of fibrosis) is possible. Hereby, the severity of liver disease and the progress of fibrosis can be determined. None of the various fibrosis marker like hyaluronate or collagen III peptid are able to replace the histology, neither alone nor as score. These biochemical marker and clinical parameter are suitable to follow up the progress of the detected fibrosis using the so-called noninvasive staging and to reduce the number of follow up biopsies. In fast progression of liver fibrosis the follow up by biopsy is almost more affirmative. For non-invasive follow ups the particular parameter hyaluronate is qualified best, according to the actual literature. Scores must keep on being evaluated on its value. PMID- 15900829 TI - [Management of chronic hepatitis B]. AB - As a leading cause of liver cirrhosis and hepatocellular carcinoma, chronic hepatitis B poses a major health care problem. Currently approved therapeutic options include interferon-alpha, pegylated interferon-alpha, lamivudine and adefovir. Interferon-alpha can induce long-term suppression of viral replication in a proportion of patients. However, treatment is associated with considerable side effects. Lamivudine and adefovir effectively suppress viral replication and induce histological improvement in the majority of patients. However, recurrence rates are high after cessation of treatment. During long-term treatment about 20% lamivudine-resistant mutants emerge annually, while currently available studies suggest that this number is about 3% for adefovir after two years of therapy. Efficacy of the respective antivirals is affected by virological and clinical parameters, thus requiring individual treatment strategies. PMID- 15900830 TI - [Eyebright. Nomen est omen]. PMID- 15900831 TI - [Investigation of methicillin resistance and aminoglycoside modifying enzyme genes in hospital staphylococci by multiplex-polymerase chain reaction]. AB - This study was conducted to investigate the presence of methicillin and aminoglycoside resistance encoding genes by multiplex-polymerase chain reaction (PCR) and by phenotypic methods in staphylococci isolated from inpatients' clinical specimens. The presence of aac(6')1aph(2"), aph(3')-IIIa and ant(4)-Ia genes encoding aminoglycoside modifying enzymes (AME) and mecA gene encoding methicillin resistance were genotypically investigated. A total of 19 S. aureus and 30 coagulase negative staphylococci (CNS) were tested. Thirty four (69.4%) of the isolates were found to be resistant to oxacillin with disk diffusion test, 33 (97%) of which were found to harbour mecA gene. The correspondance between oxacillin resistance and presence of mecA gene was found to be 100% in S. aureus isolates, while it was 95.7% in CNS. Twenty two (44.9%), 7 (14.3%) and 2 (4.1%) isolates were found to harbour aac(6')/aph(2"), aph(3')-IIIa and ant(4)-/a AME genes, respectively. At least one or more AME genes were detected in 72.7% of mecA positive isolates. PMID- 15900832 TI - [Etiological agents in bacterial meningitis and cerebrospinal fluid shunt infections and their antibiotic susceptibilities]. AB - Akut bakteriyel menenjit ve santral sinir sistemi sant enfeksiyonlarinda etkenler ve antibiyotik duyarliliklari PMID- 15900833 TI - [Inhibitor effect of vaginal lactobacilli on group B streptococci]. AB - Neonatal group B streptococcal (GBS) infections are one of the important health problems because of their high mortality and morbidity rates in certain countries. There are some preventive approaches, including perinatal antibiotic therapy against these infections. Recently, vaccination with conjugated GBS polysaccharides has also been practised. In this study, the in vitro inhibitory effects of 51 lactobacilli (of them 50 were purified from vaginal swabs, 1 from a commercial vaginal tablet) on five GBS (4 clinical isolates and 1 standard strain) were investigated by sandwich plate technique and deferred antagonism well technique. Ten clinical isolates (20%) and the drug-purified Lactobacilli expressed pronounced inhibitory effects on growth of GBS. All of the inhibitory isolates and 10 randomly selected non-inhibitory isolates were identified by API 50CHL kit (BioMeriex, France). Seven (70%) of the inhibitory clinical isolates were Lactobacillus rhamnosus. The inhibitory isolates had higher acid production than the non-inhibitory ones (p < 0.05), and pH-adjustment destroyed their inhibitory effects entirely. If these results could be applied in vivo, it could be postulated that administration of certain lactobacilli as probiotics via an appropriate regimen may be a safe, physiological and cheaper alternative for prevention of neonatal GBS infections. PMID- 15900834 TI - [Nosocomial Stenotrophomonas maltophilia infections in a university hospital]. AB - Stenotrophomonas maltophilia is a nosocomial pathogen of increasing importance. In our study, 190 S. maltophilia strains isolated from 153 hospitalized patients between January 2000-April 2004, at Farabi Hospital at Medical School of Karadeniz Technical University, were prospectively evaluated. Of these patients 67.9% were clinically compatible with nosocomial infection, and 32% were considered as colonization. It was observed that rate of infection had a tendency to increase one year of age and above 50 years of age. Nosocomial infection and/ or colonization with S. maltophilia was detected in 19.7 +/- 15.2 (1-89) days after hospitalization. The clinical manifestations were bacteremia (36.5%), pneumoniae (28.8%), urinary system infection (12.5%), surgical site infection (11.5%) and peritonitis (6.7%). The bacteremia episodes were associated with central venous catheter in 37.3% (19/51), ventilator associated pneumonia in 11.7% (6/51), urinary system infection in 7.8% (4/51), peritonitis in 3.9% (2/51), and surgical site infection in 1.9% (1/51) of cases. Nineteen patients (37.3%) had no apparent primary source of infection. Higher APACHE II score, longer duration of hospitalization and prior extended-spectrum antibiotic therapy were observed in most of the patients. Antibiotic susceptibility testing revealed that, the most effective antibiotics against the isolates were trimethoprim sulfamethoxazole (94%), ticarcillin/clavulanate (79%) and ciprofloxacin (53.5%). Crude mortality rate in the patients with S. maltophilia infections was found to be 25%. In addition, it was observed that proper antibiotic treatment had protective role against mortality (14.6% vs 63.6%; OR = 0.1, Cl95 0.12-0.42, P = 0.000). It can be concluded that to prevent infections due to S. maltophilia , effective infection control programmes and rational antibiotic use policies should be established promptly. PMID- 15900835 TI - [Retrospective evaluation of Cobas Amplicor system in the rapid diagnosis of Mycobacterium tuberculosis complex]. AB - The aim of this study was to evaluate the value of Cobas Amplicor Mycobacterium tuberculosis (MTB) polymerase chain reaction (PCR) system in the rapid diagnosis of tuberculosis. During the study period, the results of acid-fast stain (AFS), culture and Cobas Amplicor MTB system obtained from 937 clinical (158 respiratory and 779 non-respiratory) specimens were retrospectively evaluated. When culture results were accepted as the gold standard, the sensitivity, specificity, positive and negative predictive values of Cobas Amplicor MTB PCR system were found as 100% for smear-positive respiratory specimens, 75%, 91.7%, 40% and 98% for smear-negative respiratory specimens, and 89.5%, 91.8%, 65.4% and 98.1% for all of the respiratory specimens, respectively. These rates were found as follows for smear-positive, smear-negative and all of the non-respiratory specimens, respectively; 100%, 66.7%, 83.3% and 100%; 29.2%, 97.3%, 26.9% and 97.6%; and 50%, 97.1%, 44.7% and 97.6%. The overall inhibition rate for Cobas Amplicor MTB was 4.8%. In conclusion, Cobas Amplicor MTB PCR system was considered as a rapid and reliable method for the diagnosis of tuberculosis in smear positive pulmonary samples. However, in smear negative pulmonary samples and non-respiratory samples, test results should be evaluated together with clinical and other laboratory data. PMID- 15900836 TI - [Serotype distribution of enteroviruses isolated from paediatric cases prediagnosed as aseptic meningitis between 2001-2004 period]. AB - Enteroviruses have major clinical and public health importance and are one of the leading causes of aseptic meningitis. There are many diseases with similar clinical symptoms and cerebrospinal fluid (CSF) findings of aseptic meningitis, thus virus isolation and identification is crucial for definitive diagnosis. Virological diagnosis is nonetheless important to distinguish between induced meningitis and other treatable causes of disease with a similar clinical picture. A total of 249 samples obtained from 246 cases (age range: 0-15 years), prediagnosed as aseptic meningitis, were sent to Virology Laboratory of Refik Saydam Hygiene Center. The patients were followed at Department of Pediatric Infectious Diseases in the Social Security Hospital, Ankara, Turkey, between 2001 and 2004. Stool (n: 180), CSF (n: 54) and throat swab (n: 15) samples have been inoculated to RD (rhabdomyosarcoma), Hep-2 (human epithelioma) and L20B (transgenic mice) cell lines, and followed up for the presence of cytopathic effects. A total of 95 enterovirus strains were isolated from 85 (34.6%) cases, and serotyped by using RIVM (National Institute of Public and the Environment, Nederlands) antisera with microneutralization method. As a result, the most frequently isolated types were found as echovirus type 30 (n: 24) and coxsackievirus type B (n: 19), which were most frequently isolated between July to October. This is the first report from Turkey for aseptic meningitis cases due to echovirus type 25 (n:3), 18 (n:2), 14 (n:1), 13 (n:4), 11 (n:6), 9 (n:1), 6 (n:9), 5 (n:1), 4 (n:1) and coxsackievirus type A9 (n:1). PMID- 15900837 TI - [Detection of TT virus DNA by nested-PCR method in non A-E hepatitis cases]. AB - TT virus (TTV) is the unique single stranded circular DNA virus, isolated from humans. Viral DNA is shown to be present not only in patients with hepatitis of unknown etiology but also in apparently healthy populations. TTV's role as a causative agent for non A-E hepatitis is widely questioned. In this study, 23 non A-E hepatitis patients (mean age: 46.8 yrs), 21 chronic hepatitis B (CHB) patients (mean age: 41.0 yrs), 28 chronic hepatitis C (CHC) patients (mean age: 48.1yrs) have been investigated, together with 90 healthy blood donors (mean age: 33.4 yrs), for the presence of TTV-DNA by nested polymerase chain reaction (PCR). Two sets of primers targeting different regions (N22 and NCR) of the viral genome were combined to enhance the detection sensitivity. TTV-DNA was detected in 10 (43.5%) of non A-E hepatitis, 10 (35.7%) of CHC, 4 (19.1%) of CHB patients, and 16 of studied 52 (30.8%) blood donors, by N22-PCR. Viral DNA positivity rates by using NCR-PCR were found as follows for the groups respectively; 65.2% (15/23), 50% (14/28), 42.9% (9/21) and 57.8% (52/90). The differences of TTV-DNA positivity rates between study groups were found statistically insignificant, when data from each set of primer were compared (p>0.05). However, the positivity rate of non A-E hepatitis group (91.3%) was significantly higher than CHC (64.2%), CHB (47.6%), and control (57.8%) groups, when the detection sensitivities of both sets were combined (p=0.002 and p=0.046, respectively). The study also revealed that the prevalence of TTV might be underestimated if appropriate molecular methods were not employed. DNA sequencing and genotyping are required to establish TTV's role as a causative agent of non A-E hepatitis and to identify certain genotypes that might have a role in the pathogenesis of hepatitis. PMID- 15900838 TI - [Investigation of TT virus-DNA in multitransfused children and healthy children]. AB - TT virus (TTV) is a naked, single stranded DNA virus, which has been discovered in the serum of a patient with posttransfusion hepatitis of unknown etiology. TTV is widespread in the population, however, the mode of its transmission is unclear. This study was conducted to search for TTV-DNA positivity rates and its relationship with the clinical outcomes of recipients who underwent multiple blood or blood product transfusion, together with healthy children. TTV-DNA was investigated in 52 multitransfused pediatric patients (age range: 3 mnths - 17.5 yrs, mean age: 9.2 +/- 5.7 years) and 18 healthy children (age range: 1 mnth - 16.5 yrs, mean age: 8.1 +/- 4.9 years), by qualitative in-house semi-nested polymerase chain reaction (PCR) with the primers NG059, NG061 and NG063, generated from ORF1 region of the viral genome. TTV-DNA was found positive in 30.8% of multitransfused, and 16.7% of healthy children. The differences of TTV DNA positivity rates between the multitransfused and control groups, and ALT values between the patients with positive and negative TTV-DNA, were statistically insignificant (p>0.05). As a result, no relationship was detected between TTV positivity and hepatitis, although there was a statistically insignificant increase of TTV-DNA positivity in multitransfused children. However, since the primers of ORF1 N22 region used in our PCR method did not have enough sensitivity for the detection of TTV-DNA, it has been concluded that more sensitive primers such as UTR primers, should be used for more reliable evaluation of the results. PMID- 15900839 TI - [Evaluation of a commercial enzyme immunoassay for the detection of interferon gamma levels in active tuberculosis patients and vaccinated healthy subjects]. AB - The detection of plasma interferon gamma (IFN-g) levels has an important value for the evaluation of cell mediated immune response to Mycobacterium tuberculosis. The aim of this study was to investigate the plasma IFN-g levels by a commercial enzyme immunoassay (ELISA) and to compare the levels between recently diagnosed culture positive lung tuberculosis patients and BCG vaccinated healthy controls. Twenty-three patients with active lung tuberculosis (13 males, 10 females) and 34 BCG vaccinated healthy adults (16 male, 18 female) have been included in the study. The control subjects were questioned about passed tuberculosis infection and/or a contact with tuberculosis patients. No risk factors for exposure to M. tuberculosis were found in the control group. IFN-g levels were measured by QuantiFERON-TB (Cellestis, Australia) kit, and 22 of patients and 17 of control subjects were found to be positive. As a result, the sensitivity of QuantiFERON-TB test was high (95.6%), however its specificity was quite low (50%). In conclusion, QuantiFERON-TB may be used as a supplementary diagnostic test in patients considered to have active tuberculosis, before treatment. As BCG is in routine vaccination programme and the number of active tuberculosis cases is high in our country, this test seems to be invalid for the diagnosis of latent tuberculosis. Therefore, more specific tests that are not affected by the vaccine response, are required for the diagnosis of latent tuberculosis. PMID- 15900840 TI - [Bacterial contamination in blood and blood products]. AB - Skin disinfection during phlebotomy is a critical step for bacterial contamination of blood and blood products. The aim of this study was to investigate the bacterial contamination rates during phlebotomy and to detect the probable microorganisms present. Skin disinfections of 100 blood donors were performed by using povidone iodine solution with standard procedure. Fifteen mililiters of blood samples were drawn from the transfusion set and inoculated into culture flasks of automated Bact/Alert (BioMerieux) system. Blood cultures were monitorized for one week, and bacteria in positive cultures were identified by using classical microbiological methods in addition with API identification system (BioMerieux; ID32 Staph, 20 Strep). As a result, bacterial growth was detected in four (4%) of the blood samples, whereas 96% of the samples were found sterile. Staphylococcus epidermidis was the microorganism which had been grown in three of the samples, and Streptococcus mutans in one. The positivity rate detected in our study was considered high, since expected bacterial contamination rates in blood transfusions were between 0.2-0.5%. This data indicated that the procedures used in phlebotomy such as the choice of phlebotomy region, disinfectant use and disinfection time should be re-evaluated in our blood centre. PMID- 15900841 TI - [A case with fever of unknown origin during treatment for malaria: multi-drug resistant Salmonella typhi infection]. AB - Typhoid fever is an acute infectious disease caused by Salmonella serotype Typhi, leading to endemic or epidemic outbreaks in tropical/ subtropical countries (especially in India, Southeast Asia, Central and South Africa). In this report, a 27 years old male patient with malaria has been presented. The patient was diagnosed to have malaria while working in Afghanistan, and received malaria treatment since one month. He admitted to our hospital because of still continuing high fever, and other complaints (weight loss, night sweats, weakness, anorexia). His fever was 39.5 degrees C at admission, and blood smears were negative for Plasmodium sp. On the third day of admission, rose spots were detected on the skin of the abdomen and chest, and group agglutination tests gave positive results for S. Typhi O (titer: 1/800), and S. Typhi H (titer: 1/3200). Blood cultures revealed growth of Salmonella enterica serotip Typhi. The isolate was found to be resistant to ampicillin, chloramphenicol, tetracyclin and trimethoprimsulfamethoxazole, and sensitive to ciprofloxacin. The patient was treated successfully with ciprofloxacin for 14 days. PMID- 15900842 TI - [Case report: a case of tularemia with delayed diagnosis]. AB - Tularemia is a zoonotic disease caused by Francisella tularensis. In this report, a tularemia case who was living in a non-endemic area, has been presented. A 24 years old female patient with multiple cervical lymphadenitis has been admitted to the outpatient clinic of Ankara Training and Research Hospital, Turkey. Her complaints started two months ago as signs and symptoms of an upper respiratory tract infection. As she had received non-specific treatment (amoxycillin clavulanate; 2x 1g/day) ineffective against F. tularensis, lymph nodes were suppurated. The diagnosis was made serologically, microagglutination test was positive at a titer of 1/160. The antimicrobial therapy (streptomycin; 1x1 gr im, doxycycline; 2x100 mg/day) was prolonged to four weeks, according to the clinical response of suppurated lymph nodes. It can be concluded that, early diagnosis and treatment of tularemia are important to prevent abscess formation, and patients with delayed diagnosis may benefit from prolonged therapy. PMID- 15900843 TI - [Case report: cutaneous tuberculosis and tuberculous osteomyelitis]. AB - In this report, a 27 years old male patient diagnosed to have skin and bone tuberculosis (TB) has been presented. The patient admitted to the hospital with the complaints of fever, weight loss and night sweats. Patient's history revealed that following a trauma a skin lesion in the right ankle was developed and this was followed by the development of many lesions in different parts of the body. The lesions persisted despite the use of various antibiotics since a year. It has been recorded that his father has already been receiving anti-tuberculosis treatment. Osteomyelitis was detected in the distal part of right tibia by computerized tomography, and Mycobacterium tuberculosis was isolated from the specimens of skin lesion. The patient was immunocompetent, and there was no pulmonary involvement. Isoniazid (INAH), rifampin (RIF), ethambutol and morphozinamid therapy has been started and completed to 12 months with INAH and RIF. In the post-treatment follow-up of patient for one year, no relapse was detected. As a result, tuberculosis should be considered in patients with persistent skin lesions especially in endemic countries. PMID- 15900844 TI - [A transfusion-transmitted malaria case]. AB - Malaria is a rare but potentially serious complication of blood transfusion. In this report a transfusion-transmitted malaria case has been presented. A 47-years old woman admitted to our clinic with the complaints of striking fever with chills, diarrhea and vomiting. She had history of an operation and transfusion before 10 days of admission. On physical examination jaundice, splenomegaly and abdominal tenderness were detected. Laboratory results revealed anemia, and elevated LDH and bilirubin levels. Examination of thin blood films yielded Plasmodium vivax trophozoites. Chloroquine was initiated for therapy and the patient was successfully treated. On the other hand, informations about her blood donor indicated that he had been in the military service in Southeast Anatolia of Turkey where malaria is endemic. All the efforts to reach the donor, for his diagnosis and treatment, were failed. Since our region (Northeast Anatolia) is not an endemic area for malaria and the patient had no travel history to an endemic area, it has been considered that the transmission route of malaria in this case was blood transfusion. In conclusion, as there are no available approved tests for malaria screening of donations, the transfusion-transmitted malaria can only be prevented by careful questioning of the donors. PMID- 15900845 TI - [Culture based diagnostic methods for tuberculosis]. AB - Culture methods providing isolates for identification and drug susceptibility testing, still represent the gold standard for the definitive diagnosis of tuberculosis, although the delay in obtaining results still remains a problem. Traditional solid media are recommended for use along with liquid media in primary isolation of mycobacteria. At present, a number of elaborate culture systems are available commercially. They range from simple bottles and tubes such as MGIT (BD Diagnostic Systems, USA), Septi-Chek AFB (BD, USA) and MB Redox (Biotest Diagnostics, USA) to semiautomated system (BACTEC 460TB, BD, USA) and fully automated systems (BACTEC 9000 MB [BD, USA], BACTEC MGIT 960 [BD, USA], ESP Culture System II [Trek Diagnostics, USA], MB/BacT ALERT 3D System [BioMerieux, NC], TK Culture System [Salubris Inc, Turkey]). Culture methods available today are sufficient to permit laboratories to develop an algoritm that is optimal for patients and administrative needs. In this review article, the culture systems used for the diagnosis of tuberculosis, their mechanisms, advantages and disadvantages have been discussed under the light of recent literature. PMID- 15900846 TI - [Laboratory diagnosis and antibiotic susceptibilities of nontuberculous mycobacteria]. AB - Nontuberculous mycobacteria (NTM) are found widespread in nature. They can live in soil, water supplies, plants and various environmental conditions. Today there are nearly 100 known NTM species and more than 50 of them are associated with human diseases. NTM can cause pulmonary diseases, lymphadenitis, skin, soft tissue and skeletal infections, catheter related bloodstream infections and disseminated infections in patients with underlying diseases like AIDS, chronic obstructive pulmonary diseases, emphysema, bronchiectasis, cystic fibrosis and chronic alcoholism. The laboratory diagnosis of NTM has improved in parallel to the improvements in molecular biology. The new DNA sequencing and microarray technologies have been started to use in laboratories. Antibiotic susceptibility tests can be used both for taxonomic and clinical purposes. In this review article, the importance of laboratory diagnosis and antibiotic susceptibilities of NTM have been discussed. PMID- 15900847 TI - [Antibiotic resistance among enterococci isolated from clinical samples at Trakya University Hospital in the last two years]. PMID- 15900848 TI - [Antibiotic resistance of nosocomial methicillin-resistant Stahylococcus aureus isolates in our hospital]. PMID- 15900850 TI - Blood pressure guidelines and screening techniques. AB - Recording and monitoring blood pressure should be a lifetime practice. Studies have shown that people with normal blood pressure readings between the ages of 55 and 65 still have an 80 to 90 percent risk of developing hypertension by the age of 80. The information in this course teaches the dental health professional to be aware of the causes of hypertension and how to determine and assess identifiable causes. Understanding the terms systolic, diastolic and pulse pressure along with knowledge of the various blood pressure categories and the factors that influence blood pressure is important. This course provides you with the appropriate method of taking a blood pressure and how to determine the recommendations for follow-up and lifestyle modifications for patient treatment. PMID- 15900851 TI - The new roles of today's dental team. PMID- 15900852 TI - Mirror, mirror, on the wall, who's the most valuable of all? PMID- 15900853 TI - Treating the non-english speaking patient. PMID- 15900854 TI - Predictors of success on the dental assisting national board exam. PMID- 15900855 TI - Professional liability for ADAA members. PMID- 15900856 TI - Risk management assessment: dentists' ethical duty to provide emergency after hours care. PMID- 15900857 TI - Miracles in Iraq: "why you should always brush your teeth". PMID- 15900859 TI - Just a skosh ... controlling those sticky moments. PMID- 15900858 TI - Salary survey results say CDAs still earning more! PMID- 15900860 TI - Parking your car: communicating with wheels. PMID- 15900861 TI - Blood in the gutter: handling the awful call. PMID- 15900862 TI - It's care with a capital "C". PMID- 15900863 TI - The ambulance purchasing process: Part 2. PMID- 15900864 TI - Barrels of bad news. PMID- 15900865 TI - Responding to school emergencies. PMID- 15900866 TI - Wireless snapshots. PMID- 15900867 TI - The importance of rural E9-1-1. PMID- 15900868 TI - Getting there is half the battle. PMID- 15900869 TI - Mobile technology--mobile data terminals on steroids. PMID- 15900870 TI - You've got question? Research has the answers. PMID- 15900871 TI - The role of dispatch in pattern surveillance. PMID- 15900872 TI - Stings and bites: what to do about envenomation injuries. AB - Although serious envenomation injuries are rare in the United States, all emergency workers should be prepared for them. Most severe complications occur in people with previous allergies or pre-existing conditions. Many people, especially those with prediagnosed allergies, will be capable of self-treatment prior to EMS's arrival; however, prepare for the worst. EMS personnel should know how to treat virtually any envenomation injuries with general practices, and should inform the receiving hospital so venom-specific treatment can be prepared. Crews should also know what creatures are indigenous to their areas and know what to expect when the tones go off. Be aware that a side effect of any animal bite or sting, whether venomous or not, is bacterial infection. Remind patients of this. The hospital physician will also educate them about infection. If you respond to a call but the patient refuses transport and signs a waiver, you must educate him about possible bacterial contamination from the bite or sting. Never waive a patient until he has been educated regarding his injury and still refuses transport. PMID- 15900873 TI - Cocaine, excited delirium and sudden unexpected death. PMID- 15900874 TI - Strategic planning for EMS agencies. PMID- 15900875 TI - Therapeutic hypothermia. An old treatment gets a new look. PMID- 15900876 TI - Community paramedicine: part of an integrated healthcare system. PMID- 15900877 TI - Medic on the green. PMID- 15900878 TI - Monitoring the stability of Rubisco in micropropagated grapevine (Vitis vinifera L.) by two-dimensional electrophoresis. AB - Plants cultured in vitro suffer from several physiological and biochemical impairments due to the artificial conditions of growth, namely the composition of the heterotrophic media. Upon transfer to ex vitro, the higher irradiances, compared to in vitro, can lead to oxidative stress symptoms, which can be counteracted by CO2 concentrations above atmospheric levels. Here we analyse the stability of Rubisco in in vitro grapevine plantlets, and after transfer to ex vitro under four acclimatization treatments: low irradiance (LL, 150 micromol m( 2)s(-1)) and high irradiance (HL, 300 micromol m(-2)s(-1)) in association with CO2 concentrations of 350 (LCO2) and 700 (HCO2) microL L(-1). Proteins were separated with SDS polyacrylamide gel electrophoresis and two-dimensional electrophoresis and Rubisco degradation peptides were analysed by immunoblotting with anti-LSU antibodies. These degradation products were present in the leaves of plantlets under both in vitro and ex vitro treatments. Under LCO2 they were maintained for almost all of the 28 days of the acclimatization period, while becoming scarcely detected after 14 days under HCO2 and after 7 days when HCO2 was associated with HL. These results appear to confirm the counteraction of HCO2 concentrations over the oxidative stress eventually caused by HL. The patterns of soluble sugars in acclimatizing leaves under HLHCO2 also gave an indication of a faster acquisition of autotrophic characteristics. PMID- 15900879 TI - Profiling C6-C3 and C6-C1 phenolic metabolites in Cocos nucifera. AB - This paper reports the detection and identification of phenolic metabolites (C6 C3 and C6-C1 compounds) in Cocos nucifera. An HPLC/UV system was used to analyze the soluble and wall-associated phenolics in mesocarp and leaf tissues of C. nucifera. Alkaline hydrolysis of the cell wall material of the mesocarpic and leaf tissues yielded 4-hydroxybenzoic acid as the major phenolic compound. Other phenolic acids identified were ferulic acid, 4-coumaric acid, 4 hydroxybenzaldehyde and vanillic acid. No significant qualitative differences in composition were observed between leaf and mesocarp, but there were quantitative variations in the metabolite levels. PMID- 15900880 TI - Analysis of bean (Phaseolus vulgaris L.) proteins affected by copper stress. AB - The effect of excess copper on the expression of soluble proteins in 10-day old Phaseolus vulgaris seedlings was studied with two-dimensional electrophoresis and mass spectrometry, to find sensitive biochemical markers of exposure. Despite major differences in root Cu contents, both 15 and 50 microM Cu treatments resulted in equal enhancements of Cu in the primary leaves. Three proteins, apparently reacting in a dose-dependent manner to Cu exposure, were identified from roots. The levels of an intracellular pathogenesis-related protein and a newly identified protein homologous to PvPR1, PvPR2, were increased with increasing Cu concentration. The level of a newly identified PR-10 protein decreased in a dose-dependent manner. No significant difference was observed in the leaf protein pattern between controls and 15 microM Cu-treated plants. However, at 50 microM Cu exposure, the appearance of PvPR1 and a homologue of Arabidopsis thaliana thylakoid lumenal 17.4kDa protein was observed. Another protein slightly enhanced by Cu treatment had sequence homology to a mitochondrial precursor of glycine cleavage system H protein of Flaveria pringlei. PMID- 15900881 TI - Characterization of a polyclonal antiserum against the monoterpene monooxygenase, geraniol 10-hydroxylase from Catharanthus roseus. AB - Geraniol 10-hydroxylase (G10H) is a P450 containing enzyme which is the first committed step in the biosynthesis of monoterpene indole alkaloids (MIAs), including the Catharanthus roseus-anticancer drugs vinblastine and vincristine. It is thought that G10H has a regulatory role in MIA production. In the present paper, we report the characterization of a polyclonal serum raised against the purified G10H polypeptide. Anti-G10H IgG was able to inhibit the G10H activity and also recognized the G10H polypeptide from C. roseus and other plants producing MIAs. These results establish the usefulness of this antiserum as a biochemical tool for the study of G10H regulation. PMID- 15900882 TI - Antioxidant enzymes and isoflavonoids in chilled soybean (Glycine max (L.) Merr.) seedlings. AB - Changes of activity antioxidant enzymes and of levels of isoflavonoids were studied in the roots and hypocotyls of the etiolated soybean (Glycine max (L.) Merr. var. Essor) seedlings, submitted to cold. Prolonged exposure to 1 degrees C inhibited hypocotyl and root elongation and limited their growth after seedlings were transferred to 25 degrees C. Roots were more sensitive to chilling than hypocotyls. At 1 degrees C a gradual increase in MDA concentration in roots but not in hypocotyls was observed. An increase in catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1) activity in hypocotyls was observed both at 1 degrees C and after transfer of plants to 25 degrees C. In roots, CAT activity increased after 4 days of chilling, while SOD activity only after rewarming. L-Phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity decreased in roots of chilled seedlings, but did not change in hypocotyls until activity increased after transfer to 25 degrees C. The content of genistein and daidzein increased after 24 h of treatment by low temperature and then decreased with prolonged chilling in hypocotyls and remained high in roots. However, it should be noted that genistin level (genistein glucoside) in chilled hypocotyls is 10 times higher than in roots, despite falling tendency. The role of antioxidant enzymes and isoflavonoids in preventing chilling injury in hypocotyls and roots of soybean seedlings is discussed. PMID- 15900883 TI - Water deficit induced oxidative damage in tea (Camellia sinensis) plants. AB - When the tea (Camellia sinensis) leaf water potential was -1.1 MPa (Moderate water deficit), there was 58% inhibition of photosynthesis accompanied by increased zeaxanthin, malondialdehyde, oxidized proteins and superoxide dismutase activity. When the leaf water potential was -2MPa (severe water deficit), there was nearly complete inhibition of photosynthesis apart from a decrease in chlorophylls, beta-carotene, neoxanthin and lutein. Water deficit at this level caused further conversion of violaxanthin to zeaxanthin, suggesting damage to the photosynthetic apparatus. There were consistent decreases in antioxidants and pyridine nucleotides, and accumulation of catalytic Fe, malondialdehyde and oxidized proteins. It is inferred that, in tea plants, the increase in catalytic Fe and the decrease in antioxidant protection may be involved in the oxidative damage caused by severe water deficit, but not necessarily in the incipient stress induced by moderate water deficit. PMID- 15900884 TI - Uptake of sodium in quince, sugar beet, and wheat protoplasts determined by the fluorescent sodium-binding dye benzofuran isophthalate. AB - The uptake of sodium into protoplasts of quince (Cydonia oblonga Mill, clone BA29), sugar beet (Beta vulgaris L. cv. Monohill), and wheat (Triticum aestivum L. cv. Kadett) was determined by use of the acetoxy methyl ester of the fluorescent sodium-binding benzofuran isopthalate (SBFI-AM). In the presence of 1 mM CaCl2, little sodium was taken up in the cytosol of quince mesophyll cells compared to cytosols of sugar beet and wheat. Upon addition of 40 mM NaCl, approximately the same amount of sodium was taken up in leaf and root protoplasts of wheat, but no sodium was taken up in quince. However, in calcium-free medium, obtained by addition of ethylene glycol tetra acetic acid (EGTA), quince protoplasts transiently took up sodium in the cytosol when 200-400 mM NaCl was added to the protoplast medium. Moreover, after cultivation of quince in the presence of 200 mM sodium for 4 weeks, the cytosol of isolated protoplasts did not take up any sodium at all from a calcium-free medium. The results show that protoplasts from salt tolerant quince only temporarily take up sodium in the cytosol and that they have a mechanism for fast extrusion of sodium from that compartment. These mechanisms are probably important for the high salt tolerance of quince. Calcium blocks the sodium uptake into the cytosol of both quince and wheat protoplasts. PMID- 15900885 TI - Increase of photosynthesis and starch in potato under elevated CO2 is dependent on leaf age. AB - Potato plants (Solanum tuberosum cv. Bintje) were grown in open top chambers under ambient (400 microL L(-1)) and elevated CO2 (720 microL L(-1)). After 50 days one half of each group was transferred to the other CO2 concentration and the effects were studied in relation to leaf age (old, middle-aged and young leaves) in each of the four groups. Under long-term exposure to elevated CO2, photosynthesis increased between 10% and 40% compared to ambient CO2. A subsequent shift of the same plants to ambient CO2 caused a 20-40% decline in photosynthetic rate, which was most pronounced in young leaves. After shifting from long-term ambient to elevated CO2, photosynthesis also increased most strongly in young leaves (90%); these experiments show that photosynthesis was downregulated in the upper young fully expanded leaves of potato growing long term under elevated CO2. Soluble sugar content in all leaf classes under long term exposure was stable irrespective of the CO2 treatment, however under elevated CO2 young leaves showed a strongly increased starch accumulation (up to 400%). In all leaf classes starch levels dropped in response to the shift from 720 to 400 microL L(-1) approaching ambient CO2 levels. After the shift to 720 microL L(-1), sucrose and starch levels increased, principally in young Leaves. There is clear evidence that leaves of different age vary in their responses to changes in atmospheric CO2 concentration. PMID- 15900886 TI - Sap flow, gas exchange, and hydraulic conductance of young apricot trees growing under a shading net and different water supplies. AB - The experiment was carried out in a research field near Murcia, Spain, over a 3 week period between September 26 and October 16, 2000. Sixteen trees were used in the experiment, eight of which were placed under a rectangular shading net, while the other eight were maintained in the open air. Trees were irrigated once per day and, after October 5th, water was witheld from eight trees (four shaded and four unprotected for 5 days). The leaf stomatal conductance and the photosynthesis rates were higher in the shaded trees than in the exposed plants, probably because the leaf water potential was lower in the unshaded plants. This higher leaf conductance partially compensated for the effect of low radiation on transpiration, and the reduction of daily sap flow registered in shaded trees was only around 10-20%. The net also affected trunk diameter changes, with the shaded trees showing lower values of maximum daily shrinkage. Soil water deficit and high radiation had a similar effect on plant water parameters, lowering leaf water potential, leaf stomatal conductance, and the photosynthesis rate. The effects of both conditions were accumulative and so the exposed water-stressed plants showed the lowest values of total hydraulic resistance and water use efficiency, while the shaded well-irrigated trees registered the highest values for both parameters. For this reason, we think that net shading could be extended to apricot culture in many areas in which irrigation water is scarce and insolation is high. PMID- 15900887 TI - Somatic embryogenesis in Solanum tuberosum from cell suspension cultures: histological analysis and extracellular protein patterns. AB - An embryogenic cell suspension, continuously grown in Murashige and Skoog (MS) medium with 0.5 mg/L of 2,4-dichlorophenoxyacetic acid, was established from friable callus of Solanum tuberosum internode sections. The cell suspension was predominantly composed of cell masses and free embryogenic cells. When transferred to an auxin-free medium with zeatin, somatic embryos (SEs) developed and converted to complete plants when cultured on solid MS medium without growth regulators. The system produced approximately 600 SEs per 50 mL of medium. In this investigation, accumulation of extracellular proteins (EPs) of different molecular weights were found associated to different phases of the embryogenic process. At the initiation of the cell suspension, cell clusters and free cells present in the culture (phase "A") secreted a 78kDa EP, unique to this phase. In phase "B", which is related to embryonic cell determination process, proteins (7 14kDa) were secreted mainly by embryogenic cells. In phase "C", SEs in different developmental stages secreted protein of 32 kDa, which appeared as a particular feature of the phase. EPs of phase "D", secreted by torpedo and mature embryos, had molecular weights between 20 and 50 kDa. Further studies will be necessary to identify these proteins and link them to previously identified somatic embryogenesis-related proteins. Histological analysis of the potato embryogenesis in liquid media showed unicellular origin of the SE. PMID- 15900888 TI - An anther-specific cysteine-rich protein of tomato localized to the tapetum and microspores. AB - The tapetum is a nutritive tissue of the stamen that is essential for normal microspore development. While numerous tapetal-specific genes have been identified, little information is available on the localization and function of the proteins produced by these genes. The tapetally produced protein 5B-CRP is cysteine-rich, has a secretory signal sequence and lacks an endoplasmic reticulum retention sequence. The 5B-CRP mRNA is expressed specifically within the tapetum and accumulates from premeiosis to tetrad release. Antibodies generated against an Escherichia coli fusion protein only recognized 5B-CRP in the reduced state. The 5B-CRP was detected as a 6 kDa protein in extracts of stamens from microspore meiosis through anthesis and was also observed in extracts from dehisced pollen. In situ, 5B-CRP was localized in stamens to the tapetum and the developing microspores, from the tetrad through early free microspore stages. Based on similarity to proteins with known functions, 5B-CRP may inhibit proteasome activity within the stamen locule. PMID- 15900889 TI - Enhanced drought tolerance of transgenic rice plants expressing a pea manganese superoxide dismutase. AB - We investigated the role that manganese superoxide dismutase (MnSOD), an important antioxidant enzyme, may play in the drought tolerance of rice. MnSOD from pea (Pisum sativum) under the control of an oxidative stress-inducible SWPA2 promoter was introduced into chloroplasts of rice (Oryza sativa) by Agrobacterium mediated transformation to develop drought-tolerant rice plants. Functional expression of the pea MnSOD in transgenic rice plants (T1) was revealed under drought stress induced by polyethylene glycol (PEG) 6000. After PEG treatment the transgenic leaf slices showed reduced electrolyte leakage compared to wild type (WT) leaf slices, whether they were exposed to methyl viologen (MV) or not, suggesting that transgenic plants were more resistant to MV- or PEG-induced oxidative stress. Transgenic plants also exhibited less injury, measured by net photosynthetic rate, when treated with PEG. Our data suggest that SOD is a critical component of the ROS scavenging system in plant chloroplasts and that the expression of MnSOD can improve drought tolerance in rice. PMID- 15900890 TI - Micropropagation of Dendrobium nobile from shoot tip sections. AB - Successful shoot regeneration of Dendrobium nobile was achieved using thin shoot tip sections and triacontanol (TRIA) for the first time. Protocorm-like bodies (PLBs) or proliferating shoot buds were observed when thin shoot tip sections were cultured on the basal medium of Mitra et at. (Indian J. Exp. Biol. 14 (1976) 350) supplemented with 4.0 microg L(-1) TRIA. The highest percentage of explants (93%) produced PLBs or proliferating shoot buds (21) at 4.0 microg L(-1) TRIA supplemented basal medium. All the newly formed PLBs or proliferating shoot buds survived and ultimately produced healthy shoots with 2-3 leaves. Shoots produced roots when cultured on basal medium supplemented with 2.0 microg L(-1) TRIA. The well-rooted shoots were transferred to pots containing charcoal chips, coconut husk and broken tiles (2:2:1), and a 92% survival rate was achieved. This work reveals that TRIA can be used as an effective growth regulator in the micropropagation and conservation of D. nobile. PMID- 15900891 TI - cDNA cloning and expression of the phytoene synthase gene in sunflower. AB - A PCR approach was used to isolate a phytoene synthase (Psy) cDNA from sunflower (Helianthus annuus L.). A reconstructed full-length sequence (1598 bp) of the Psy cDNA was obtained; it contained a 1242 bp CDS, 172-nucleotides of 5'-untranslated region (UTR), and 170-nucleotides of 3'-UTR. The predicted protein (46.8kDa) displayed a sequence of 414 amino acid residues with a putative transit sequence for plastid targeting in the N-terminal region. Phylogenetic analysis demonstrated that the sunflower Psy (HaPsy) clustered with the marigold (Tagetes erecta) Psy gene, with which it showed an overall amino acid identity of 93.7%. Moreover, the HaPsy sequence relates closely with other Psy sequences of higher plants. HaPsy was highly expressed in cotyledons, and young and mature leaves. In contrast, HaPsy transcript levels were comparatively lower in the stem and almost absent in the roots. The HaPsy transcript levels were influenced by leaf expansion, which suggested that the expression of the HaPsy gene is regulated during the process of leaf development. The role of HaPsy in controlling carotenoid biosynthesis is demonstrated by the concurrent increase of HaPsy transcript levels with the light-dependent enhanced carotenoid production in green tissues of sunflower. PMID- 15900892 TI - Third meeting of the Cysticercosis Working Group in Eastern and Southern Africa takes place in Maputo. PMID- 15900893 TI - Cardiac involvement in canine babesiosis. AB - Cardiac dysfunction in canine babesiosis has traditionally been regarded as a rare complication, with the majority of lesions reported as incidental findings at post-mortem examination. Recent studies have, however, demonstrated cardiac lesions in canine babesiosis. Cardiac troponins, especially troponin I, are sensitive markers of myocardial injury in canine babesiosis, and the magnitude of elevation of plasma troponin I concentrations appears to be proportional to the severity of the disease. ECG changes in babesiosis are similar to the pattern described for myocarditis and myocardial ischaemia and together with histopathological findings indicate that the heart suffers from the same pathological processes described in other organs in canine babesiosis, namely inflammation and hypoxia. The clinical application of the ECG appears to be limited and thus cardiovascular assessment should be based on functional monitoring rather than an ECG tracing. On cardiac histopathology from dogs that succumbed to babesiosis, haemorrhage, necrosis, inflammation and fibrin microthrombi in the myocardium were documented, all of which would have resulted in ECG changes and elevations in cardiac troponin. Myocardial damage causes left ventricular failure, which will result in hypotension and an expansion of the plasma volume due to homeostatic mechanisms. PMID- 15900894 TI - Epidemiology and control of trematode infections in cattle in Zimbabwe: a review. AB - In this paper the main epidemiological aspects of the major domestic ruminant trematode infections in Zimbabwe are reviewed and discussed with regard to the available options for control. Seasonal occurrence of amphistomes, Fasciola gigantica and Schistosoma mattheei are considered both in the definitive and intermediate hosts. The regional distribution of the trematodes is reviewed in relation to the distribution patterns of their snail intermediate hosts. Based on the epidemiological features of the trematodes, practical control measures are suggested. PMID- 15900895 TI - Anaesthesia of nyala (Tragelaphus angasi) with a combination of thiafentanil (A3080), medetomidine and ketamine. AB - A combination of thiafentanil (A3080), medetomidine hydrochloride (MED) and ketamine hydrochloride (KET) was evaluated in 19 boma-habituated (12 female and 7 males) and 9 free-ranging nyala (7 male and 2 females) (Tragelaphus angasi) to develop a safe and reliable anaesthesia protocol. Wide dosages were used safely during this study with ranges for A3080 of 45 +/- 8 microg/kg with MED of 69 +/- 19 microg/kg and KET of 3.7 +/- 1.0 mg/kg (200 mg/ animal). The dosages developed on boma-habituated nyala proved to be equally effective in 9 adult free-ranging nyala (7 males and 2 females). The optimum dosage for nyala was a combination of A3080 (40-50 microg/kg), MED (60-80 microg/kg) plus 200 mg of KET/animal. The anaesthesia was characterised by a short induction, good muscle relaxation and mild hypoxaemia during monitoring the anaesthesia was rapidly and completely reversed by naltrexone hydrochloride (30 mg/mg of A3080) and atipamezole hydrochloride (5 mg/mg of MED) given intramuscularly. There was no mortality or morbidity associated with this protocol. PMID- 15900896 TI - Effect of mastitis on macro-minerals of bovine milk and blood serum in Sudan. AB - Milk and blood serum from clinically mastitis infected, subclinically mastitis infected and healthy Friesian cows (15 samples from each of 3 groups) were evaluated for macrominerals (sodium, potassium, calcium, magnesium and phosphorus). The milk from cows infected with subclinical mastitis revealed a significant decrease in potassium (P < 0.001) and a significant increase in sodium and phosphorus content (P < 0.01). Similarly, the milk from cows with the clinical form of the disease showed a significant increase in sodium (P < 0.001) and a significant decrease in potassium, magnesium (P < 0.001) and calcium (P < 0.01). Comparison of healthy cow's milk with that from cows with subclinical mastitis revealed a highly significant increase in sodium (P < 0.001). Comparison of healthy cow's milk with that of clinically mastitic milk showed a highly significant decrease in levels of calcium, magnesium (P < 0.001) and potassium (P < 0.01). However, sodium increased highly significantly (P < 0.001). Comparison of macro-minerals in milk from cows with subclinical and clinical mastitis revealed a significant decrease in potassium contents (P < 0.05) compared with that of healthy cows. Potassium levels were found to decrease significantly (P < 0.05) in subclinically infected cow's blood serum. However, calcium and phosphorus showed a significant decrease (P < 0.01) in blood serum samples from the clinically infected cows. PMID- 15900897 TI - Clinical and clinicopathological changes in 6 healthy ponies following intramuscular administration of multiple doses of imidocarb dipropionate. AB - Haematological variables and selected serum indices, particularly those affected by changes in renal and hepatic function, were examined in 6 healthy ponies following 4 intramuscular doses of 4 mg/kg imidocarb dipropionate administered every 72 hours. This treatment regime has been reported to sterilise experimental Babesia equi infections in horses and may have value in preventing the spread of this disease during exportation of possible carrier horses to non-endemic countries. Serum bile acids and serum gamma glutamyltransferase activity were measured to evaluate the effect of this treatment regime on hepatic function. Owing to the absence of any increase in these variables it was concluded that this treatment regime had no clinically detectable deleterious effect on hepatic function in healthy ponies. Urinary gamma glutamyltransferase : creatinine ratios (IU/g), serum creatinine and fractional clearance of sodium, potassium and phosphate (%) were calculated as a measure of renal function. Urinary GGT and urinary GGT : creatinine ratios were significantly elevated on Day 5 of the trial, with 2 of the trial animals also exhibiting mild azotaemia indicative of changes in renal function. The changes in urine GGT : urine creatinine ratios observed in this study also provides evidence of the value of this ratio for the early detection of renal toxicity, following exposure to nephrotoxic agents. PMID- 15900898 TI - The effect of water intake prior to blood sampling on packed cell volume in sheep. AB - The effect of water intake prior to blood sampling on subsequent packed cell volume (PCV) was investigated in ewe lambs (8 months of age) of the Dohne Merino, Merino and Letelle flocks at Grootfontein Agricultural Development Institute. On the afternoon of the day before the experiment was conducted, a blood sample for a baseline PCV value (R) was taken from each animal. The following day, 15 ewes of each breed (n = 45) were dosed with 1 litre of water and another 15 of each (n = 45) were used as a control. Blood samples for PCV were taken concurrently for both the control and water treatment groups before the dose (0), and at 15, 30 and 60 minutes after dosing. PCV were subsequently determined with the microcapillary centrifuge technique. Baseline PCV of Letelle ewes was higher (32.4 +/- 0.6) than that of the Dohne Merino (29.7 +/- 0.6) and Merino (28.7 +/- 0.6) ewes. Furthermore, recovery rate at 30 minutes after treatment also differed among breeds. Although there were significant differences between the control and water treatment groups at R and 0 minutes, which were probably due to inherent animal differences, there were no significant differences between PCV of the 2 groups during the remainder of the experimental period. Overall it can be concluded that water intake before blood sampling for the determination of PCV has no significant effect on haematocrit. Differences among breeds were more pronounced than those between treatment groups. PMID- 15900899 TI - Post-anaesthetic myelomalacia in a horse. AB - This article describes a rare neurological complication of anaesthesia in a 2 year-old Clydesdale colt undergoing castration. Anaesthesia was induced with glyceryl guaiacol ether and ketamine and maintained with halothane. Following an uneventful anaesthetic of 40 minutes, the horse recovered from anaesthesia in a padded recovery stall. After approximately 70 minutes in the recovery stall, the horse attempted to stand and adopted a dog sitting position. One hundred and fifty minutes later, the horse became distressed and was sedated with xylazine. Clinical examination of the horse did not reveal any evidence of myositis or fractures. A neurological examination revealed an intact anal reflex, deep pain response in the hind legs, tail tone and voluntary movement of the hind legs was possible. The horse deteriorated neurologically over the next 24 hours and was euthanased on humane grounds. The horse was submitted for necropsy. Gross pathology was unremarkable except for a small amount of haemorrhage around the right kidney. Histopathology revealed no abnormalities in any muscle groups or peripheral nerves. Congestion and axonal swelling of the spinal cord was evident from T16 to S1. Ischaemic neurons were evident from L 1 to L 6. The most prominent lesions were at L4 and L5. A diagnosis of myelomalacia was made. This is a rare complication of anaesthesia in horses with 9 case studies appearing in the literature since 1979. This is the 1st case to be reported in South Africa. The speculated pathophysiology and risk factors for this complication are discussed. PMID- 15900900 TI - Aeromonas hydrophila-associated skin lesions and septicaemia in a Nile crocodile (Crocodylus niloticus). AB - Aeromonas hydrophila is one of the most common bacteria associated with the aquatic environment. There are, however, limited data on A. hydrophila infection in crocodilians. The aim of this report is to describe a case of skin lesions and septicaemia associated with A. hydrophila in a Nile crocodile (Crocodylus niloticus). A captive male crocodile in the Zoological Park of Antalya (Turkey) was found dead without showing signs of any disease. Gross examination showed brown or red-spotted skin lesions of varying size. These lesions were mostly scattered over the abdomen and occasionally on the tail and feet. At necropsy, numerous white, multifocal and randomly distributed areas were seen on the liver. Gram-stained smears from skin and liver lesions showed Gram-negative bacilli arranged in clusters. Pure cultures of A. hydrophila were recovered from skin, internal organs and blood. Isolates were found to be susceptible to ceftiofur, amoxicillin + clavulanic acid, oxytetracycline, enrofloxacin, danofloxacin, neomycin, gentamicin, and lincomycin + neomycin. A pathogenicity test was performed using this isolate on 4 male 2-year-old New Zealand white rabbits. Local abscesses formed in 2 rabbits injected subcutaneously and the 2 that were injected intraperitoneally died as a result of septicaemia. In conclusion, this report has shown that A. hydrophila may cause skin lesions and even death due to septicaemia in crocodiles. PMID- 15900901 TI - Papillary ovarian cystadenocarcinoma in a dog. AB - An 11-year-old female German Shepherd dog was presented for investigation of progressive enlargement of the abdomen, periodic bloody discharge from the vulva and rapid exhaustion. Transabdominal ultrasonography and lateral abdominal radiography demonstrated an echogenic formation with anechogenic cavities located cranial to the urinary bladder and a homogeneous shadow with an elliptical shape was located caudal to the rib arc. Both showed indistinct borders. Exploratory laparotomy identified bilateral ovarian masses and ovariohysterectomy was performed. Histopathology confirmed ovarian cystadenocarcinoma. The dog remained clinically normal without evidence of metastatic disease 4 months after surgery. Papillary cystadenocarcinoma in the bitch could affect both ovaries and manifests with a rapid growth rate and clinical signs such as rapid exhaustion, abdominal enlargement and vulval discharge. Ovariohysterectomy is the treatment option. PMID- 15900902 TI - Small intestinal foreign body in an adult Eclectus parrot (Eclectus roratus). AB - A 14-month-old female Eclectus parrot (Eclectus roratus) was presented with a 4 week history of bloody diarrhea and depression. No additional information could be gained from physical examination. Only selected diagnostic tests (faecal examination, haematocrit, aspartate aminotransferase, and uric acid) could be performed due to financial constraints, but all where within reference range. Unspecific antibiotic treatment was started and the bird responded well initially, but had to be readmitted 2.5 weeks after initial presentation. Four weeks after initial presentation the owner finally consented to taking whole body radiographs and a diagnosis of an intestinal foreign body could be made. The foreign body was surgically removed 2 days later. The bird recovered uneventfully after surgery and was still in good health 1 year after surgery. This article emphasises the importance of diagnostic imaging in the avian patient. A brief review of avian gastrointestinal foreign bodies is given (concentrating on the psittacine patient) and the importance of distinguishing metallic from non metallic gastrointestinal foreign bodies are discussed. PMID- 15900903 TI - Tumor vaccine: current trends in antigen specific immunotherapy. AB - Effective cancer treatment to prevent the tumor growth as well as to stop its recurrence is the dream of oncologists. Currently available therapeutic measures like, radiotherapy and chemotherapy, often suffer from severe toxicity and lack of specificity of the drug towards tumor cells. Another promising approach is the 'immunotherapy', in which either the immune system is activated by tumor vaccine to combat the tumor growth or antitumor antibodies can be used. Vaccination can stimulate humoral, cellular and innate immune systems to generate various effector molecules, like antibody, cytotoxic T cells, cytokines etc. In antigen specific immunotherapy, the immune system can be stimulated actively by antigen based tumor vaccine to kill only those tumor cells, having expression of the particular tumor associated antigen. Different experimental, preclinical and clinical studies have proved that generated immune responses are effective to restrict the tumor growth. Useful strategies of antigen specific immunotherapy and outcome of various laboratory and clinic based studies are discussed. PMID- 15900904 TI - Effect of glutamine analogue-acivicin on tumor induced angiogenesis in Ehrlich ascites carcinoma. AB - The inhibition of tumor growth and tumor induced angiogenesis by the glutamine antimetabolite acivicin was evaluated in 6-7 weeks old male Swiss albino mice bearing Ehrlich ascites carcinoma (EAC) transplanted by intraperitoneal (ip) injections of EAC cells. Treatment involving ip injections with two different doses of acivicin (0.05 and 0.41microg/g body weight/day) in saline revealed decrease in tumor volumes and reduced number of blood vessels on peritoneal wall after 10 and 15 days of treatment when compared to control (i.e. injected with saline only). Vascular hyperpermeability was found to be lesser in the treated groups of mice than the control as indicated by the FITC- D and colloidal carbon assay. Serum VEGF level was found to decrease in the drug treated groups both after 10 and 15 days of treatment. The results thus suggest that acivicin may suppress tumoral angiogenesis through regulation of VEGF level. PMID- 15900905 TI - Sheep erythrocyte demonstrated better effect than IL-2 and IFN-gamma as biological response modifier against glioma in experimental model. AB - The significant insights into the immunobiology of central nervous system (CNS) and brain tumor have opened up the feasibility of applying 'Immunotherapy' as an alternative to the poor prognosis of malignant brain tumor with conventional therapeutic approaches. Though cytokines like IL-2 and IFN-gamma used against glioma showed some favorable results by eliciting Th1 type immune response, a proper immunotherapeutic agent is still to be searched for. Sheep erythrocyte (SRBC), a corpuscular antigen showed a better therapeutic efficacy in terms of enhanced survival and augmentation of cell mediated immunity (CMI) in a glioma model developed by chemical carcinogen ethyl nitrosourea. Histological findings revealed most efficient glioma rejection in SRBC and combination biological response modifier (BRM) treated groups. Simultaneously E-rosetting, cytotoxicity of lymphocytes, phagocytosis and antigen presenting capacity of myeloid cells established the better therapeutic efficacy of SRBC alone than other BRMs viz. IL 2 and IFN-gamma. Even the effect of combination therapy of different BRMs showed marginal differences in facilitating glioma reduction than the single use of SRBC. These findings emphasized the application of SRBC as an exogenous BRM having the potential as a rational therapeutic adjunct against glioma. PMID- 15900906 TI - Anti-lipidperoxidative role of exogenous dehydroepiendrosterone (DHEA) administration in normal ageing rat brain. AB - Effects of exogenous dehydroepiendrosterone (DHEA) administration on the levels of lipid proxidation products, malondialdyde (MDA)-a thiobarbuteric acid reactive substance (TBARS) and 4-hydroxynonenal (4-HNE) in different brain regions viz. cerebral cortex, hippocampus cerebellum, and brain stem of 12 and 22 months old rats were studied. DHEA treatment significantly depressed TBARS and 4-HNE in all the brain regions studied, in both the age group rats. Interestingly, the magnitude of decrease was higher in the 22 months old rats than that in 12 months old rats. The results suggest that older the animal, better will be the response of exogenous DHEA administration against age-related peroxidative products. PMID- 15900907 TI - Acute analgesic effect of loperamide as compared to morphine after intrathecal administration in rat. AB - Loperamide, a mu opioid receptor agonist, which is commonly used as an antidiarrhoeal agent has been reported to possess analgesic activity after intrathecal administration. However, the exact analgesic profile, i.e., onset, duration and intensity of analgesia in relation to morphine is not fully known. In the present study, the acute analgesic effect of loperamide (5 microg) was compared with that of morphine (5 microg) and morphine + loperamide (5 microg of each) using the tail flick method after intrathecal administration. Naloxone (5 mg/kg) reversibility of the analgesic effect was also studied. The analgesic response of loperamide was significantly higher than morphine. Even after 22 hr, maximum possible effect was greater than 49%. Naloxone partially antagonized the analgesic effect of loperamide. This suggested that loperamide may be acting through blockade of Ca2+ channels besides activating mu opioid receptors. Loperamide may prove to be a better substitute for morphine as spinal analgesic. PMID- 15900908 TI - Effect of Emblica officinalis (Gaertn) on CCl4 induced hepatic toxicity and DNA synthesis in Wistar rats. AB - A single dose of CCl4 (1 ml/kg body weight, po in corn oil) increased the levels of SGOT (serum glutamate oxaloacetate transaminase), SGPT (serum glutamate pyruvate transaminase), LDH (lactate dehydrogenase), glutathione-S-transferase and depletion in reduced glutathione, glutathione peroxidase and glutathione reductase. It also caused enhancement in the levels of lipid peroxidation (LPO) and DNA synthesis. There was also pathological deterioration of hepatic tissue as evident from multivacuolated hepatocytes containing fat globules around central vein. The pretreatment of E. officinalis for 7 consecutive days showed a profound pathological protection to liver cell as depicted by univacuolated hepatocytes. Pretreatment with E. officinalis at doses of 100 and 200 mg/kg body weight, prior to CCl4 intoxication showed significant reduction in the levels of SGOT, SGPT, LDH, glutathione-S-transferase, LPO and DNA synthesis. There was also increase in reduced glutathione, glutathione peroxidase and glutathione reductase. The results suggest that E. officinalis inhibits hepatic toxicity in Wistar rats. PMID- 15900909 TI - Nutraceutical effects of garlic oil, its nonpolar fraction and a Ficus flavonoid as compared to vitamin E in CCl4 induced liver damage in rats. AB - Carbon tetrachloride feeding (3.2g/kg/72hr) for one month increased significantly the serum and tissue lipid profile and deranged the enzyme levels viz; alkaline phosphatase, alanine transaminase, aspartate transaminase, glutathionze reductase, HMGCoA reductase, catalase, gluc.6.PDH and malic enzyme in rats. Simultaneously the lipid peroxidation level in liver was also raised. On administration of garlic oil and its major nonpolar fraction (NPFGO) and a flavonoid isolated from the bark of Ficus bengalensis Linn, viz; leucopelargonin derivative respectively to different groups(100mg/kg/day) the deleterious effects of CCl4 were significantly ameliorated. The liver damage by CCl4 was satisfactorily prevented by these samples as effectively as Vit. E (50 mg/kg/day). The results prove that important nutraceuticals (phytonutrients) like bioflavonoids and theols i.e. allylic sulphide rich fractions give protection from toxins like CCl4. The order of beneficial effects of the drugs are Leucopelargonin > NPFGO > Garlic oil and their effects are comparable to that of vitamin E used at a minimal dose. PMID- 15900910 TI - Effect of chronic ethanol administration on testicular antioxidant system and steroidogenic enzyme activity in rats. AB - In order to find out the effect of chronic ethanol administration on testicular antioxidant system and steroidogenic enzyme activity, male rats fed with ethanol 1.6g/kg body weight per day for four weeks were studied. Besides a drastic reduction in body and testis weight, there was decrease in ascorbic acid, reduced glutathione and activities of superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase in the testicular tissue of the treated animals. Simultaneously, there was increase in lipid peroxidation and glutathione S-transferase activity. Activities of 3 beta-hydroxy steroid dehydrogenase and 17 beta-hydroxy steroid dehydrogenase were also found decreased in the treated animals. The results indicate that chronic ethanol administration resulted in increase in oxidative stress and decrease in the activities of steroidogenic enzymes in the rat testes. PMID- 15900911 TI - Effect of sublethal levels of nitrite on some blood parameters of juvenile Labeo rohita (Hamilton-buchanan). AB - Juveniles of L. rohita were exposed to sublethal levels of nitrite (0.02, 0.1 and 0.4 mg/l) for 2, 24, 48 and 96 hr. The time of exposure at individual concentrations of nitrite did not show any significant difference in haemoglobin, cortisol, chloride and lactic acid. Haematocrit showed significant reduction with increasing concentration of nitrite irrespective of duration of exposure. Fishes exposed to 0.4 mg/l nitrite showed significantly high levels of glucose beyond 2 hr. The mean erythrocytic fragility of fishes exposed to the 3 concentrations of nitrite for 3 exposure periods showed significant higher sensitivity to osmotic stress. The results suggest decrease in haematocrit and cell wall strength of erythrocytes creating stress to fish. PMID- 15900912 TI - Genetic basis of hybrid male sterility among three closely related species of Drosophila. AB - The genetic basis of hybrid male sterility among three closely related species, Drosophila bipectinata, D. parabipectinata and D. malerkotliana has been investigated by using backcross analysis methods. The role of Y chromosome, major hybrid sterility (MHS) genes (genetic factors) and cytoplasm (non-genetic factor) have been studied in the hybrids of these three species. In the species pair, bipectinata--parabipectinata, Y chromosome introgression of parabipectinata in the genomic background of bipectinata and the reciprocal Y chromosome introgression were unsuccessful as all males in second backcross generation were sterile. Neither MHS genes nor cytoplasm was found important for sterility. This suggests the involvement of X-Y, X-autosomes or polygenic interactions in hybrid male sterility. In bipectinata--malerkotliana and parabipectinata--malerkotliana species pairs, Y chromosome substitution in reciprocal crosses did not affect male fertility. Backcross analyses also show no involvement of MHS genes or cytoplasm in hybrid male sterility in these two species pairs. Therefore, X- autosome interaction or polygenic interaction is supposed to be involved in hybrid male sterility in these two species pairs. These findings also provide evidence that even in closely related species, genetic interactions underlying hybrid male sterility may vary. PMID- 15900913 TI - Molecular tagging of gene conferring leaf blight resistance using microsatellites in sorghum [Sorghum bicolor (L.) Moench]. AB - Resistance to leaf blight in sorghum [Sorghum bicolor (L.) Moench] accession G 118 was found to segregate as a single dominant trait in a cross to susceptible cultivar, HC-136. Molecular marker(s) linked to the locus for disease resistance was identified using simple sequence repeat (SSR) markers coupled with bulk segregant analysis. Genomic DNA from the parental cultivars and bulks were screened by PCR amplification with 50 simple sequence repeat primer pairs. Out of these, 38 SSR primers produced polymorphism between parents. After screening of these 38 SSRs with resistant and susceptible bulk, one SSR primer, Xtxp 309 produced a unique band of approximately 700 bp only in resistant parent and resistant bulk and a unique band of 450 bp only in susceptible parent and susceptible bulk. Upon screening with individual resistant and susceptible recombinant inbred lines (RILs), marker Xtxp 309 produced amplification in 23 of the 26 resistant RILs and no amplification was produced in any of the 25 susceptible RILs. The same marker Xtxp 309 produced amplification in 21 of the susceptible RILs and 3 of the resistant RILs of 450 bp band. This was found to be located at a distance of 3.12 cM away from the locus governing resistance to leaf blight which was considered to be closely linked and 7.95 cM away from the locus governing susceptibility to leaf blight. This marker may prove useful in MAS for gene introgression, plant genetic diagnostics and gene pyramiding for resistance via genetic transformation for disease resistance in plants. PMID- 15900914 TI - Characterization of an exopolysaccharide produced by a marine Enterobacter cloacae. AB - An exopolysaccharide producing marine bacterium, Enterobacter cloacae, was isolated from marine sediment collected from Gujarat coast, India. Chemical investigation of exopolysaccharide (EPS 71 a) revealed that this exopolysaccharide was an acidic polysaccliaride containing high amount of uronic acid, fucose and sulfate which is rare for bacterial exopolysaccharides. EPS 71a was found to have fucose, galactose, glucose and glucuronic acid in a molar ratio of 2: 1: 1: 1. PMID- 15900915 TI - Phytopesticidal and repellent efficacy of Litsea salicifolia (Lauraceae) against Aedes aegypti and Culex quinquefasciatus. AB - Litsea salicifolia is one of the many plants used as phytopesticide, traditionally by various tribes of Assam. Of the five extracts of L. salicifolia tested for the bioactivity against Aedes aegypti and Culex quinquefasciatus, the aqueous extract was more effective compared to other extracts exhibiting bioactivity at 72 ppm against A. aegypti. The hexane extract (2000 ppm) exhibited 70% repellent activity for 3 hr against A. aegypti and 46% activity for 3 hr against C. quinquefasciatus. This is the first report on the insecticidal properties of L. salicifolia which can be further developed as an eco-friendly biopesticides of the future. PMID- 15900916 TI - Comparative protein profiles of Salmonella and Escherichia coli. AB - Protein profiles of selected Salmonella serovars were compared with E. coli to identify genus specific protein(s) for Salmonella. The PDP formed of different Salmonella serovars were compared with E. coli O78 when subjected to SDS-PAGE yielded 11, 15, 15, 11 and 14 bands in S. Bareilly, S. Gallinarum, S. Typhimurium and S. Weltevreden and E. coli O78 respectively. The bands produced were compared with each other. It was found that S. Weltevreden shared 7 bands with E. coli O78, A protein of molecular weight 20.89 kDa was found in all Salmonella serovars, but not in E. coli O78 suggesting its genus specific attribute. PMID- 15900917 TI - Retrieving text inferences: controlled and automatic influences. AB - Bridging inferences contribute to text coherence by identifying the connections among ideas, whereas elaborative inferences simply specify sensible extrapolations from text. Bridging inferences have been indistinguishable from explicit text ideas on numerous measures, suggesting similar longterm memory (LTM) representations for the two, whereas elaborative inferences are inferior. To evaluate the LTM representations of text ideas, we used the extended process dissociation procedure (Buchner, Erdfelder, & Vaterrodt-Plunnecke, 1995; Jacoby, 1991) to partition the controlled, recollective contributions to text retrieval from the automatic, familiarity-based contributions. The automatic contribution to the recognition of implied concepts was consistently negligible, an outcome consistent with the absence of perceptual processing of those concepts during the original reading. In addition, the controlled basis of recognition was consistently higher for explicit than for implicit concepts, which suggests a more robust conceptual representation for explicit text ideas (Yonelinas, 2002). These results were interpreted to reflect the asymmetric representation of explicit ideas and inferences (elaborative and even bridging inferences) in the surface, propositional textbase, and situational levels of text representation. PMID- 15900918 TI - Text cohesion and metacomprehension: immediate and delayed judgments. AB - In three experiments, we examined comprehension judgments made after a piece of text had been read. We propose that such metacognitive judgments are based on the content of working memory at the exact moment of assessment. Generally speaking, this working metacognition hypothesis is in agreement with Koriat's cue utilization approach, which implies that different elements of information will be available (and used) depending on the moment at which a judgment is made. More specifically, our hypothesis focuses on the management of working memory during reading as a cause for cue (un)availability. In support of these views, the results of Experiment 1 showed that a cohesion manipulation affecting the comprehension of specific paragraphs influences judgments only on these paragraphs, and not on judgments on the whole text. In Experiment 2, we showed that an interfering task that takes place just before this paragraph judgment is made wipes out this cohesion effect. Experiment 3 showed, on the other hand, that the whole-text judgment may, nevertheless, be affected by text cohesion, provided that the readers keep an access to the text when the judgment is made. These results support the idea that working memory management makes different cues available for metacognitive ratings at different delays. PMID- 15900919 TI - Modulation of regularity and lexicality effects in reading aloud. AB - We examined the question of whether the sizes of the regularity and lexicality effects in naming can be modulated as a function of filler type (nonwords or low frequency exception words). The lexicality effect was larger in the exception word filler condition than in the nonword filler condition, but the size of the regularity effect was essentially unaffected by filler type. This pattern is at odds with what is generally assumed to be the predictions from dual-route theories of reading aloud. An attempt was next made to determine whether the dual route cascaded model of Coltheart, Rastle, Perry, Langdon, and Ziegler (2001) could possibly simulate this pattern when changes were introduced to each of the three parameters that affect the contribution of the nonlexical route. We discuss the implications of these results for the idea that reliance on the lexical and nonlexical routes is under strategic control. PMID- 15900920 TI - The retrieval practice effect in associative recognition. AB - Recalling an item interferes with recall of related memories. Evidence is presented that retrieval interference occurs in associative recognition as well as recall. In Experiment 1, subjects studied pairs of category exemplars. Retrieval practice followed, during which some pairs appeared in a cued recall test. A final test of associative recognition (with remember-know judgments) found lower accuracy and hit rate for nonpracticed pairs belonging to retrieval practiced categories. In Experiment 2, subjects studied noun pairs from overlapping sets, with study duration manipulated between subjects. Retrieval practice was manipulated by presenting some members of a set in a previous block during the recognition test. With long study duration, retrieval interference was evident in both recognition and remember judgments. With short study duration, it appeared only in remember judgments. These results support a dual-process account in which retrieval interference is specific to recollection and becomes evident in recognition performance only when recollection is sufficiently dominant. PMID- 15900921 TI - Associative interference in recognition memory: a dual-process account. AB - Associative interference from overlapping word pairs (A-B, A-D) reduces recall but has inconsistent effects on recognition. A dual-process account suggests that interference conditions reduce recollection but increase familiarity. This is predicted to increase recognition false alarms but have variable effects on recognition hits, depending on the relative contribution of recollection and familiarity. In three experiments that varied materials (sentences or random nouns) and test type (associative or pair recognition), interference conditions always increased recognition false alarms, but sometimes increased and sometimes decreased recognition hits. However, remember hits always decreased and know hits always increased with interference, patterns predicted of the recollection and familiarity processes, respectively. According to the dual-process view, a manipulation that affects the component processes in opposite ways can produce inconsistent patterns of recognition performance as the relative contribution of recollection and familiarity changes across tasks. PMID- 15900922 TI - Pairs do not suffer interference from other types of pairs or single items in associative recognition. AB - What is the source of interference on a memory test following study of a list containing different types of pairs? Many current models predict that pairs and singles of all types will jointly interfere and therefore harm memory. Such list length effects have often been observed for lists of a single-item type (e.g., a list of words). Here, we examine interference for lists containing multiple types of pairs (e.g., word-word, face-face, word-face). In three experiments, we manipulate the number of each type on the study list. In associative recognition, discrimination fell as the number of pairs of the same type rose, but the number of pairs of other types had little effect. That is, we found a list length effect within, but not between, classes of stimuli. We highlight the importance of representation and propose alternatives to current model representations that can predict such findings. PMID- 15900923 TI - Time versus items in judgment of recency. AB - In the numerical judgment of recency (JOR) task, subjects judge how many items have intervened since the test item was previously presented. Two experiments were conducted to determine whether the basis of JOR is the age of the memory (time) or the number of intervening items. Subjects went through a long list that was made up of alternating fast blocks and slow blocks, but the block structure was disguised by probabilistic selection of a short or long intertrial interval. In both experiments, JOR was found to be a simple function of time, with no added contribution from the number of items. PMID- 15900924 TI - Are first impressions lasting impressions? An exploration of the generality of the primacy effect in memory for repetitions. AB - In five experiments, we investigated the primacy effect in memory for repetitions (DiGirolamo & Hintzman, 1997), the finding that when participants are shown a study list that contains two very similar versions of the same stimulus, memory is biased in the direction of the version that was presented first. In the experiments reported, the generality of the effect was examined by manipulating the orientation and features of the repeated stimuli. The results confirmed that the effect is reliable when stimulus changes affect the accidental properties of the stimulus (properties of the stimulus that give information about distance or angle but do little to aid in identification). However, the effect was not found when changes were made to other aspects of the stimulus. The results suggest that the primacy effect in memory for repetitions is not robust across all stimulus changes and converge with previous findings that have demonstrated that such properties of stimuli as orientation and size are represented differently in memory than are other stimulus characteristics. PMID- 15900925 TI - Intention to learn influences the word frequency effect in recall but not in recognition memory. AB - Watkins, LeCompte, and Kim (2000) suggested that the recall advantage for rare words in mixed lists is due to a compensatory study strategy that favors the rare words. They found the advantage was reversed when rare and common words were studied under incidental learning conditions. The present study investigated the possibility that the rare-word advantage in recognition memory is also the result of a compensatory study strategy. Experiment 1 replicated the findings of Watkins et al. that the rare-word advantage in recall is eliminated under incidental learning conditions. In contrast, Experiment 2 showed that the rare-word advantage in recognition memory is maintained under both intentional and incidental learning conditions. Experiment 3 replicated the results of Experiments 1 and 2 using different stimuli and a different orienting task. Finally, Experiment 4 showed that the rare-word advantage in recognition is maintained with pure lists. These findings show that the rare-word advantage in recognition memory is not the result of a compensatory study strategy. Instead, rare words are encoded more distinctively than common words, irrespective of participants' intention to remember them. PMID- 15900926 TI - Visual short-term memory is not improved by training. AB - A critical question in visual working or short-term memory (VSTM) research is whether the ability to remember briefly presented visual stimuli can be increased. Here we test whether VSTM for locations and shapes is improved by training that allows one to utilize another memory system, visual longterm memory (VLTM). Training was done by repeatedly presenting a subset of memory displays, creating long-term memory traces for these displays. Surprisingly, VSTM performance for repeated displays was not higher than for nonrepeated ones, even though participants recognized repeated displays on a forced-choice test given at the end of the experiment. We suggest that the fidelity of information held by VLTM is inferior to that of information held by VSTM and thus provides no additional benefit over what is extracted on the fly by VSTM. PMID- 15900927 TI - Age-related differences in executive control of working memory. AB - In two experiments, we used dual-task methodology to assess the effect of aging on executive control of working memory. We hypothesized that (1) age-related dual task costs would be observed even when individual tasks represent different perceptual modalities; (2) age would modulate the effect of increased temporal overlap on dual-task performance; and (3) the vulnerability of specific memory mechanisms to interference would be age related. We found that aging was associated with disproportionate dual-task costs that increased when extending the overlap between individual tasks. The effect of interference with encoding, and arguably output, was disproportionately larger in old than in young individuals. Ensuring that individual tasks represent different perceptual modalities is important but insufficient when using dual-task methodology to assess the effect of aging on executive function. The degree of temporal overlap between individual tasks and the sensitivity of specific memory operations to interference should be considered, as well. PMID- 15900928 TI - Conversation and convention: enduring influences on name choice for common objects. AB - The name chosen for an object is influenced by both short-term history (e.g., speaker-addressee pacts) and long-term history (e.g., the language's naming pattern for the domain). But these influences must somehow be linked. We propose that names adopted through speaker-addressee collaboration have influences that carry beyond the original context. To test this hypothesis, we adapted the standard referential communication task. The first director of each matching session was a confederate who introduced one of two possible names for each object. The director role then rotated to naive participants. The participants later rated name preference for the introduced and alternative names for each object. They also rated object typicality or similarity to each named category. The name that was initially introduced influenced later name use and preference, even for participants who had not heard the name from the original director. Typicality and similarity showed lesser effects from the names originally introduced. Name associations built in one context appear to influence retrieval and use of names in other contexts, but they have reduced impact on nonlinguistic object knowledge. These results support the notion that stable conventions for object names within a linguistic community may arise from local interactions, and they demonstrate how different populations of speakers may come to have a shared understanding of objects' nonlinguistic properties but different naming patterns. PMID- 15900929 TI - Learning categories by making predictions: an investigation of indirect category learning. AB - Categories are learned in many ways, but studies of category learning have generally focused on classification learning. This focus may limit the understanding of categorization processes. Two experiments were conducted in which participants learned categories of animals by predicting how much food each animal would eat. We refer to this as indirect category learning, because the task andthe feedback were not directly related to category membership, yet category learning was necessary for good performance in the task. In the first experiment, we compared the performance of participants who learned the categories indirectly with the performance of participants who first learned to classify the objects. In the second experiment, we replicated the basic findings and examined attention to different features during the learning task. In both experiments, participants who learned in the prediction-only condition displayed a broader distribution of attention than participants who learned in the classification-and-prediction condition did. Some participants in the prediction only group learned the family resemblance structure of the categories, even when a perfect criterial attribute was present. In contrast, participants who first learned to classify the objects tended to learn the criterial attribute. PMID- 15900930 TI - The hot hand fallacy and the gambler's fallacy: two faces of subjective randomness? AB - The representativeness heuristic has been invoked to explain two opposing expectations--that random sequences will exhibit positive recency (the hot hand fallacy) and that they will exhibit negative recency (the gambler's fallacy). We propose alternative accounts for these two expectations: (1) The hot hand fallacy arises from the experience of characteristic positive recency in serial fluctuations in human performance. (2) The gambler's fallacy results from the experience of characteristic negative recency in sequences of natural events, akin to sampling without replacement. Experiment 1 demonstrates negative recency in subjects' expectations for random binary outcomes from a roulette game, simultaneously with positive recency in expectations for another statistically identical sequence-the successes and failures of their predictions for the random outcomes. These findings fit our proposal but are problematic for the representativeness account. Experiment 2 demonstrates that sequence recency influences attributions that human performance or chance generated the sequence. PMID- 15900931 TI - Goal specificity and knowledge acquisition in statistics problem solving: evidence for attentional focus. AB - Solving training problems with nonspecific goals (NG; i.e., solving for all possible unknown values) often results in better transfer than solving training problems with standard goals (SG; i.e., solving for one particular unknown value). In this study, we evaluated an attentional focus explanation of the goal specificity effect. According to the attentional focus view, solving NG problems causes attention to be directed to local relations among successive problem states, whereas solving SG problems causes attention to be directed to relations between the various problem states and the goal state. Attention to the former is thought to enhance structural knowledge about the problem domain and thus promote transfer. Results supported this view because structurally different transfer problems were solved faster following NG training than following SG training. Moreover, structural knowledge representations revealed more links depicting local relations following NG training and more links to the training goal following SG training. As predicted, these effects were obtained only by domain novices. PMID- 15900932 TI - Fictive motion as cognitive simulation. AB - Sentences such as The road runs through the valley and The mountain range goes from Canada to Mexico include a motion verb but express no explicit motion or state change. It is argued that these sentences involve fictive motion, an implicit type of motion. But do people trying to understand these sentences mentally simulate motion? This question was addressed in four experiments. In each, participants read a story about travel--for instance, fast versus slow, short versus long distance, and easy versus difficult terrain--and then made a timed decision about a fictive motion sentence. Overall, latencies were shorter after they had read about fast travel, short distances, and easy terrains. Critically, the effect did not arise with nonfictive motion target sentences (e.g., The road is in the valley), as was demonstrated in three control studies. The results suggest that the processing of fictive motion includes mental simulation. PMID- 15900933 TI - Effect of exercise on age-related changes in collagen fibril diameter distributions in the common digital extensor tendons of young horses. AB - OBJECTIVE: To determine whether specific treadmill exercise regimens would accelerate age-related changes in collagen fibril diameter distributions in the common digital extensor tendon (CDET) of the forelimbs of young Thoroughbreds. ANIMALS: 24 female Thoroughbreds. PROCEDURE: Horses were trained for 18 weeks (6 horses; short term) or 18 months (5 horses; long term) on a high-speed treadmill; 2 age-matched control groups (6 horses/group) performed walking exercise only. Horses were (mean +/- SD) 24 +/- 1 months and 39 +/- 1 months old at termination of the short-term and long-term regimens, respectively. Midmetacarpal CDET specimens were obtained and processed for transmission electron microscopy. Diameter and area of at least 1,000 collagen fibrils/specimen were measured by use of computerized image analysis. Mass-average diameter (MAD) of collagen fibrils and collagen fibril index were calculated for each horse. RESULTS: Collagen fibril MAD for the older horses was significantly less than that for the younger horses. Exercise did not significantly affect fibril diameter or distributions in either age group, and collagen fibril index did not differ significantly between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Age-related reduction in collagen fibril MAD agreed with findings for other tendons and species. Training did not accelerate age-related change in the CDET in contrast to a reported decrease in collagen fibril MAD in the superficial digital flexor tendon of horses trained long term. Our results support the concept that the functionally distinct nature of the CDET and superficial digital flexor tendon in horses results in fundamentally different responses to high-speed exercise regimens. PMID- 15900934 TI - Effects of caffeine on exercise performance of physically fit Thoroughbreds. AB - OBJECTIVE: To determine the effects of a dose of caffeine (2.5 mg/kg, IV) administered to physically fit Thoroughbreds during incremental exercise testing to fatigue on a treadmill. ANIMALS: 10 conditioned Thoroughbreds. PROCEDURE: Horses were randomly assigned to receive caffeine or a control solution. Each horse received both treatments in a crossover design with a 3-week interval between treatments. Each horse was administered caffeine (2.5 mg/kg) or an equivalent amount of a control solution IV. One hour after injection, each horse performed an incremental exercise test to exhaustion. Hematologic values, heart rate, oxygen consumption, carbon dioxide production, plasma lactate concentration, urine and serum concentrations of caffeine and metabolites, and time until exhaustion were monitored. Statistical analysis was performed by use of a mixed-effects linear model. RESULTS: Significant differences in measured values when horses were treated with caffeine or the control solution were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: A dose of caffeine (2.5 mg/kg, IV) appears to have no effect on any performance variable of physically fit Thoroughbreds during incremental exercise testing to fatigue. PMID- 15900935 TI - Influence of general anesthesia on pharmacokinetics of intravenous lidocaine infusion in horses. AB - OBJECTIVE: To compare the disposition of lidocaine administered IV in awake and anesthetized horses. ANIMALS: 16 horses. PROCEDURE: After instrumentation and collection of baseline data, lidocaine (loading infusion, 1.3 mg/kg administered during 15 minutes (87 microg/kg/min); constant rate infusion, 50 microg/kg/min) was administered IV to awake or anesthetized horses for a total of 105 minutes. Blood samples were collected at fixed times during the loading and maintenance infusion periods and after the infusion period for analysis of serum lidocaine concentrations by use of liquid chromatography with mass spectral detection. Selected cardiopulmonary parameters including heart rate (HR), mean arterial pressure (MAP), arterial pH, PaCO2, and PaO2 were also recorded at fixed time points during lidocaine administration. Serum lidocaine concentrations were evaluated by use of standard noncompartmental analysis. RESULTS: Serum lidocaine concentrations were higher in anesthetized than awake horses at all time points during lidocaine administration. Serum lidocaine concentrations reached peak values during the loading infusion in both groups (1,849 +/- 385 ng/mL and 3,348 +/- 602 ng/mL in awake and anesthetized horses, respectively). Most lidocaine pharmacokinetic variables also differed between groups. Differences in cardiopulmonary variables were predictable; for example, HR and MAP were lower and PaO2 was higher in anesthetized than awake horses but within reference ranges reported for horses under similar conditions. CONCLUSIONS AND CLINICAL RELEVANCE: Anesthesia has an influence on the disposition of lidocaine in horses, and a change in dosing during anesthesia should be considered. PMID- 15900936 TI - Evaluation of transdermal application of glipizide in a pluronic lecithin gel to healthy cats. AB - OBJECTIVE: To evaluate plasma glipizide concentration and its relationship to plasma glucose and serum insulin concentrations in healthy cats administered glipizide orally or transdermally. ANIMALS-15 healthy adult laboratory-raised cats. PROCEDURE: Cats were randomly assigned to 2 treatment groups (5 mg of glipizide, PO or transdermally) and a control group. Blood samples were collected 0, 10, 20, 30, 45, 60, 90, and 120 minutes and 4, 6, 10, 14, 18, and 24 hours after administration to determine concentrations of insulin, glucose, and glipizide. RESULTS: Glipizide was detected in all treated cats. Mean +/- SD transdermal absorption was 20 +/- 14% of oral absorption. Mean maximum glipizide concentration was reached 5.0 +/- 3.5 hours after oral and 16.0 +/- 4.5 hours after transdermal administration. Elimination half-life was variable (16.8 +/- 12 hours orally and 15.5 +/- 15.3 hours transdermally). Plasma glucose concentrations decreased in all treated cats, compared with concentrations in control cats. Plasma glucose concentrations were significantly lower 2 to 6 hours after oral administration, compared with after transdermal application; concentrations were similar between treatment groups and significantly lower than for control cats 10 to 24 hours after treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Transdermal absorption of glipizide was low and inconsistent, but analysis of our results indicated that it did affect plasma glucose concentrations. Transdermal administration of glipizide is not equivalent to oral administration. Formulation, absorption, and stability studies are required before clinical analysis can be performed. Transdermal administration of glipizide cannot be recommended for clinical use at this time. PMID- 15900937 TI - Relationships between hoof-acceleration patterns of galloping horses and dynamic properties of the track. AB - OBJECTIVE: To define relationships between hoof-acceleration patterns of galloping horses and dynamic properties of the track. ANIMALS: 8 Thoroughbred horses without lameness. PROCEDURE: Acceleration-time curves were recorded by use of accelerometers attached to each hoof as each horse galloped over the track straightaway. Four sessions were conducted for each horse, with the track surface modified by sequentially adding water before each session. These acceleration time curves were analyzed to determine peak accelerations during the support phase of the stride. Track dynamic properties (hardness, rebound, deceleration rate, rebound rate, and penetration) were recorded with a track-testing device. Moisture content and dry density were measured from soil samples. Stepwise multiple regression was used to identify relationships between hoof-acceleration variables and track dynamic properties. RESULTS: Track rebound rate was most consistently related to hoof variables, especially through an inverse relationship with negative acceleration peaks for all hooves. Also, rebound rate was related to initial acceleration peak during propulsion of the hooves of the forelimb and the nonlead hind limb as well as to the second acceleration peak during propulsion of the lead hooves of the hind limb and nonlead forelimb. CONCLUSIONS AND CLINICAL RELEVANCE: The inverse relationship between track rebound rate and negative acceleration peaks for all hooves reflects the most important dynamic property of a track. Any factor that reduces negative acceleration of the hooves will increase stride efficiency by allowing smoother transition from retardation to propulsion and therefore may be important in determining the safety of racing surfaces. PMID- 15900938 TI - Tracheobronchoscopic assessment of exercise-induced pulmonary hemorrhage in horses. AB - OBJECTIVE: To determine the interobserver variability of assessment of exercise induced pulmonary hemorrhage (EIPH) during tracheobronchoscopic examination in horses. ANIMALS: 747 Thoroughbred racehorses. Procedures-850 tracheobronchoscopic examinations were performed within 2 hours of racing for the horses. Examinations were recorded on videotape, and EIPH and its severity were assessed independently by 3 veterinarians. Concordance was determined by calculation of the Cohen weighted kappa statistic and tabulation of scores assigned by each observer. RESULTS: Weighted kappa statistics ranged from 0.75 to 0.80. In 99.4% of observations, all observers agreed or 2 of 3 agreed and the third differed by < or = 1 grade. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that interobserver reliability of tracheobronchoscopic assessment of EIPH in Thoroughbred racehorses is high when the examination is conducted by experienced veterinarians. Concordance among investigators is sufficient to justify use of this grading system for further studies and clinical descriptions of EIPH. PMID- 15900939 TI - Evaluation of vacuum and dynamic cell seeding of polyglycolic acid and chitosan scaffolds for cartilage engineering. AB - OBJECTIVES: To compare combined vacuum and rotation with the spinner flask technique for seeding chondrocytes on chitosan versus polyglycolic acid matrices. SAMPLE POPULATION: Porcine chondrocytes. PROCEDURE: A suspension containing 5 X 10(6) chondrocytes/scaffold was used to evaluate 2 seeding techniques, including a spinner flask and a custom-designed vacuum chamber used for 2 hours prior to transfer to a bioreactor. For each seeding technique, prewetted scaffolds were composed of polyglycolic acid (PGA) mesh or macroporous chitosan sponge. Constructs were collected at 48 hours for DNA quantification, measurement of water and gycosaminoglycan (GAG) content, and scanning electron microscopy. RESULTS: Yield of both seeding techniques was similar for each type of scaffold. Percentage of cells contained in the center of PGA constructs was increased with seeding in the bioreactor (43% of total cell number), compared with the spinner flask (18%). The DNA content and cell number per construct were 10 times greater for PGA constructs, compared with chitosan constructs. Chitosan scaffolds seeded in the bioreactor yielded a significantly higher GAG:DNA ratio than did PGA scaffolds. Whereas chondrones formed on chitosan scaffolds, cell distribution was more uniform on PGA scaffolds. CONCLUSIONS AND CLINICAL RELEVANCE: The vacuum bioreactor technique allowed seeded chondrocytes to attach to PGA scaffolds within 48 hours and improved uniformity of cell distribution, compared with the spinner technique. Although formation of extracellular matrix may be stimulated by seeding chitosan scaffolds in the bioreactor, further evaluations of the seeding technique and characteristics of chitosan scaffolds are warranted. PMID- 15900940 TI - Effects of sevoflurane dose and mode of ventilation on cardiopulmonary function and blood biochemical variables in horses. AB - OBJECTIVE: To quantitate effects of dose of sevoflurane and mode of ventilation on cardiovascular and respiratory function in horses and identify changes in serum biochemical values associated with sevoflurane anesthesia. ANIMALS: 6 healthy adult horses. PROCEDURE: Horses were anesthetized twice: first, to determine the minimum alveolar concentration (MAC) of sevoflurane and second, to characterize cardiopulmonary and serum biochemical responses of horses to 1.0, 1.5, and 1.75 MAC multiples of sevoflurane during controlled and spontaneous ventilation. Results-Mean (+/- SEM) MAC of sevoflurane was 2.84 +/- 0.16%. Cardiovascular performance during anesthesia decreased as sevoflurane increased; the magnitude of cardiovascular depression was more severe during mechanical ventilation, compared with spontaneous ventilation. Serum inorganic fluoride concentration increased to a peak of 50.8 +/- 7.1 micromol/L at the end of anesthesia. Serum creatinine concentration and sorbitol dehydrogenase activity reached their greatest values (2.0 +/- 0.8 mg/dL and 10.2 +/- 1.8 U/L, respectively) at 1 hour after anesthesia and then returned to baseline by 1 day after anesthesia. Serum creatine kinase, aspartate aminotransferase, and alkaline phosphatase activities reached peak values by the first (ie, creatine kinase) or second (ie, aspartate aminotransferase and alkaline phosphatase) day after anesthesia. CONCLUSIONS AND CLINICAL RELEVANCE: Sevoflurane causes dose-related cardiopulmonary depression, and mode of ventilation further impacts the magnitude of this depression. Except for serum inorganic fluoride concentration, quantitative alterations in serum biochemical indices of liver- and muscle-cell disruption and kidney function were considered clinically unremarkable and similar to results from comparable studies of other inhalation anesthetics. PMID- 15900941 TI - Virulence of fungal spores determined by tracheal inoculation of goats following inhalation of aerosolized sterile feedyard dust. AB - OBJECTIVE: To compare the virulence of spores of 7 fungi by tracheal inoculation of goats following exposure of goats to an aerosol of sterilized feedyard dust. Animals-54 weanling Boer-Spanish goats. PROCEDURE: A prospective randomized controlled study was conducted. There were 7 fungal treatment groups, a tent control group, and a pen control group (n = 6 goats/group). Goats in the 7 treatment and tent control groups were exposed to autoclaved aerosolized feedyard dust for 4 hours in a specially constructed tent. Goats in the 7 treatment groups were then inoculated intratracheally with 30 mL of a fungal spore preparation, whereas tent control goats were intratracheally inoculated with 30 mL of physiologic saline (0.9% NaCI) solution. These treatments were repeated each week for 6 weeks. RESULTS: Severity of pathologic changes differed significantly among the 7 fungal treatment groups as determined on the basis of gross atelectatic and consolidated lung lesions and histologic lesions of the lungs. Descending order for severity of lesions was Mucor ramosissimus, Trichoderma viride, Chaetomium globosum, Stachybotrys chartarum, Aspergillus fumigatus, Penicillium chrysogenum, and Monotospora lanuginosa. Trichoderma viride spores were the most invasive and were isolated from the bronchial lymph nodes and thoracic fluid of all 6 goats administered this organism. Spores were observed-histologically in lung tissues harvested 72 hours after inoculation from all treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: 4 of 7 fungal spore types induced significantly larger lung lesions, compared with those induced by the other 3 spore types or those evident in control goats. PMID- 15900942 TI - Evaluation of displacement of the digital cushion in response to vertical loading in equine forelimbs. AB - OBJECTIVE: To evaluate patterns of digital cushion (DC) displacement that occur in response to vertical loading of the distal portion of the forelimb in horses. Sample Population-Forelimbs from 10 horses with normal feet. PROCEDURE: Patterns of DC displacement induced by in vitro vertical limb loading were determined. Load-induced displacement of the DC was defined as the magnitude and direction of displacement of 6 radio-dense, percutaneously implanted markers in specific regions of the DC. The effects of solar support and nonsupport on displacement of the DC were compared. RESULTS: Regional displacement of the DC occurred principally along distal and palmar vectors in response to vertical loading. Medial or lateral abaxial displacements were variable and appeared to be dependent on response of the limb to the applied load. Displacement of the DC was not affected by the degree of solar support. CONCLUSIONS AND CLINICAL RELEVANCE: Data indicated that the biomechanical function of the DC is to act as a restraint to the displacement of the second phalanx or as a passive structure that allows flexibility of the caudal two thirds of the foot. Results did not indicate that the DC provides a force that induces displacement of or an active restraint against outward displacement of the hoof wall capsule. PMID- 15900943 TI - Digital blood flow and plasma endothelin concentration in clinically endotoxemic horses. AB - OBJECTIVE: To measure plasma endothelin-1 (ET-1) concentrations and digital blood flow in clinically endotoxemic horses. ANIMALS: 36 adult horses that underwent emergency celiotomy for primary gastrointestinal tract disease. PROCEDURE: On days 2 and 5 following surgery, Doppler ultrasonographic digital arterial blood flow measurements were obtained. Hematologic and biochemical analyses were performed, and plasma concentrations of ET-1 and endotoxin (lipopolysaccharide) were determined. A scoring system based on 9 clinical variables was used to assign horses to group B (quartile with greatest cumulative score) or group A (remaining 3 quartiles). Follow-up at 2.5 years was obtained by telephone questionnaire. RESULTS: For all horses on day 2, median (interquartile values) plasma ET-1 concentrations were 1.4 (0.8, 1.7) pg/mL, whereas on day 5, plasma ET 1 concentrations were 1.0 (0.5, 1.6) pg/mL. On day 2, digital blood flow was 0.057 (0.02, 0.07) mL/min in group A horses and 0.035 (0.02, 0.03) mL/min in group B horses. On day 5, plasma ET-1 concentration was significantly (73%) higher in group B horses, compared with group A horses. Thirty of 36 horses were alive at 2.5 years; group A horses were more likely to have survived (odds ratio, 25; 95% confidence interval, 2.4 to 262). Significant associations were found between an increase in digital pulses, hoof wall temperatures, or both and increased digital blood flow (0.14 vs 0.04 mL/min) on day 2 and increased digital arterial diameter (0.32 vs 0.23 cm) on day 5. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with more severe endotoxemia had decreased digital blood flow, increased plasma ET-1 concentrations, and decreased long-term survival. PMID- 15900944 TI - Hemodynamic effects of orally administered carvedilol in healthy conscious dogs. AB - OBJECTIVE: To evaluate the hemodynamic effects of orally administered carvedilol in healthy dogs with doses that might be used to initiate treatment in dogs with congestive heart failure. ANIMALS: 24 healthy dogs. PROCEDURE: Dogs were randomly allocated to receive carvedilol PO at a dose of 1.56, 3.125, or 12.5 mg, twice daily for 7 to 10 days; 6 dogs served as controls. Investigators were blinded to group assignment. Hemodynamic variables were recorded prior to administration of the drug on day 1 and then 2, 4, and 6 hours after the morning dose on day 1 and days 7 to 10. Change in heart rate after IV administration of 1microg of isoproterenol/kg and change in systemic arterial blood pressure after IV administration of 8 microg of phenylephrine/kg were recorded 2 and 6 hours after administration of carvedilol. RESULTS: Administration of carvedilol did not significantly affect resting hemodynamic variables or response to phenylephrine. The interaction of day and carvedilol dose had a significant effect on resting heart rate, but a significant main effect of carvedilol dose on resting heart rate was not detected. Increasing carvedilol dose resulted in a significant linear decrease in heart rate response to isoproterenol. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy conscious dogs, orally administered carvedilol at mean doses from 0.08 to 0.54 mg/kg given twice daily did not affect resting hemodynamics. Over the dose range evaluated, there was a dose-dependent attenuation of the response to isoproterenol, which provided evidence of beta adrenergic receptor antagonism. PMID- 15900945 TI - Effect of an indwelling nasogastric tube on gastric emptying rates of liquids in horses. AB - OBJECTIVE: To evaluate the effect of an indwelling nasogastric tube on gastric emptying of liquids in horses. ANIMALS: 9 healthy adult horses. PROCEDURE: A randomized block crossover design was used. For treatment group horses, a nasogastric tube was placed and 18 hours later, acetaminophen was administered; the nasogastric tube remained in place until the experiment was complete. For control group horses, a nasogastric tube was passed into t stomach, acetaminophen was administered, and the nasogastric tube was removed immediately. Serial blood samples were collected 15 minutes before and after administration of acetaminophen. Serum concentration of acetaminophen was determined by use of fluorescence polarization immunoassay. The variables, time to maximum acetaminophen concentration (Tmax) and the appearance constant for acetaminophen (Kapp), were determined. The values for Kapp and Tmax in horses with and without prolonged nasogastric tube placement were compared. RESULTS: No significant difference was found in Kapp between horses with and without prolonged nasogastric tube placement; the median difference in Kapp was 0.01 min(-1) (range, -0.48 to 0.80 min(-1). No significant difference was found in Tmax between horses with and without prolonged nasogastric tube placement; the median difference in Tmax was 5 minutes (range, -30 to 50 minutes). Reanalysis of data following the removal of possible outlier values from 1 horse resulted in a significant difference in Tmax between horses with and without prolonged nasogastric tube placement. CONCLUSIONS AND CLINICAL RELEVANCE: Although no clinically important impact of 18 hours of nasogastric intubation was found on gastric emptying in healthy was found among horses. PMID- 15900946 TI - Computer-assisted kinematic evaluation of induced compensatory movements resembling lameness in horses trotting on a treadmill. AB - OBJECTIVE: To characterize compensatory movements of the head and pelvis that resemble lameness in horses. ANIMALS: 17 adult horses. PROCEDURE: Kinematic evaluations were performed while horses trotted on a treadmill before and after shoe-induced lameness. Lameness was quantified and the affected limb determined by algorithms that measured asymmetry in vertical movement of the head and pelvis. Induced primary lameness and compensatory movements resembling lameness were assessed by the Friedman test. Association between induced lameness and compensatory movements was examined by regression analysis. RESULTS: Compensatory movements resembling lameness in the ipsilateral forelimb were seen with induced lameness of a hind limb. There was less downward and less upward head movement during and after the stance phase of the ipsilateral forelimb. Doubling the severity of lameness in the hind limb increased severity of the compensatory movements in the ipsilateral forelimb by 50%. Compensatory movements resembling lameness of the hind limb were seen after induced lameness in a forelimb. There was less upward movement of the pelvis after the stance phase of the contralateral hind limb and, to a lesser extent, less downward movement of the pelvis during the stance phase of the ipsilateral hind limb. Doubling the severity of lameness in the forelimb increased compensatory movements of the contralateral hind limb by 5%. CONCLUSIONS AND CLINICAL RELEVANCE: Induced lameness in a hind limb causes prominent compensatory movements resembling lameness in the ipsilateral forelimb. Induced lameness in a forelimb causes slight compensatory movements resembling lameness in the ipsilateral and contralateral hind limbs. PMID- 15900947 TI - Assessment of the effect of dilution of blood samples with sodium heparin on blood gas, electrolyte, and lactate measurements in dogs. AB - OBJECTIVE: To evaluate the effect of dilution of blood samples with sodium heparin on blood gas, electrolyte, and lactate measurements in dogs. Sample Population-Venous blood samples collected from 6 adult dogs of various breeds. PROCEDURE: Syringes were prepared with anticoagulant via 1 of 4 techniques, and the residual volume of liquid heparin in each type of prepared syringe was determined. Blood gas values and other selected clinicopathologic variables were measured in whole blood samples after collection (baseline) and after aliquots of the samples were diluted with heparin via 1 of the 4 manual syringe techniques. By use of a tonometer, whole blood samples were adjusted to 1 of 3 oxygen concentrations (40, 100, or 600 mm Hg) and the PO2 values were measured at baseline and subsequent to the 4 heparin dilutions. RESULTS: The 4 syringe techniques resulted in 3.9%, 9.4%, 18.8%, and 34.1% dilutions of a 1-mL blood sample. Compared with baseline values, dilution of blood samples with liquid heparin significantly changed the measured values of PCO2, PO2, and base deficit and concentrations of electrolytes and lactate. Of the variables assessed, measurement of ionized calcium concentration in blood was most affected by heparin dilution. CONCLUSIONS AND CLINICAL RELEVANCE: These findings in dogs indicate that dilution of blood samples with heparin can be a source of preanalytical error in blood gas, electrolyte, and lactate measurements. Limiting dilution of blood samples with heparin to < 4% by volume via an evacuation technique of syringe heparinization is recommended. PMID- 15900948 TI - Assessment of the hemodynamic effects of lidocaine administered IV in isoflurane anesthetized cats. AB - OBJECTIVE: To determine the hemodynamic effects of lidocaine (administered IV to achieve 6 plasma concentrations) in isoflurane-anesthetized cats. ANIMALS: 6 cats. PROCEDURE: Cats were anesthetized with isoflurane in oxygen (end-tidal isoflurane concentration set at 1.25 times the predetermined individual minimum alveolar concentration). Lidocaine was administered IV to each cat to achieve target pseudo-steady-state plasma concentrations of 0, 3, 5, 7 9, and 11 microg/mL, and isoflurane concentration was reduced to an equipotent concentration. At each plasma lidocaine concentration, cardiovascular and blood gas variables; PCV; and plasma total protein, lactate, lidocaine, and monoethylglycinexylidide concentrations were measured in cats before and during noxious stimulation. Derived variables were calculated. RESULTS: n isoflurane anesthetized cats, heart rate, cardiac index, stroke index, right ventricular stroke work index, plasma total protein concentration, mixed-venous PO2 and hemoglobin oxygen saturation, arterial and mixed-venous bicarbonate concentrations, and oxygen delivery were significantly lower during lidocaine administration, compared with values determined without lidocaine administration. Mean arterial pressure, central venous pressure, pulmonary artery pressure, systemic and pulmonary vascular resistance indices, PCV, arterial and mixed venous hemoglobin concentrations, plasma lactate concentration, arterial oxygen concentration, and oxygen extraction ratio were significantly higher during administration of lidocaine, compared with values determined without lidocaine administration. Noxious stimulation did not significantly affect most variables. CONCLUSIONS AND CLINICAL RELEVANCE: In isoflurane-anesthetized cats, although IV administration of lidocaine significantly decreased inhalant requirements, it appeared to be associated with greater cardiovascular depression than an equipotent dose of isoflurane alone. Administration of lidocaine to reduce isoflurane requirements is not recommended in cats. PMID- 15900949 TI - Effects of desflurane and mode of ventilation on cardiovascular and respiratory functions and clinicopathologic variables in horses. AB - OBJECTIVE: To quantitate the effects of desflurane and mode of ventilation on cardiovascular and respiratory functions and identify changes in selected clinicopathologic variables and serum fluoride values associated with desflurane anesthesia in horses. ANIMALS: 6 healthy adult horses. PROCEDURE: Horses were anesthetized on 2 occasions: first, to determine the minimum alveolar concentration (MAC) of desflurane in O2 and second, to characterize cardiopulmonary and clinicopathologic responses to 1X, 1.5X, and 1.75X desflurane MAC during both controlled and spontaneous ventilation. RESULTS: Mean +/- SEM MAC of desflurane in horses was 8.06 +/- 0.41 %; inhalation of desflurane did not appear to cause airway irritation. During spontaneous ventilation, mean PaCO2 was 69 mm Hg. Arterial blood pressure, stroke volume, and cardiac output decreased as the dose of desflurane increased. Conditions of intermittent positive pressure ventilation and eucapnia resulted in further cardiovascular depression. Horses recovered quickly from anesthesia with little transient or no clinicopathologic evidence of adverse effects. Serum fluoride concentration before and after administration of desflurane was below the limit of detection of 0.05 ppm (2.63microM/L). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that desflurane, like other inhalation anesthetics, causes profound hypoventilation in horses. The magnitude of cardiovascular depression is related to dose and mode of ventilation; cardiovascular depression is less severe at doses of 1X to 1.5X MAC, compared with known effects of other inhalation anesthetics under similar conditions. Desflurane is not metabolized to an important degree and does not appear to prominently influence renal function or hepatic cellular integrity or function. PMID- 15900950 TI - Effect of a ventral slot procedure and of smooth or positive-profile threaded pins with polymethylmethacrylate fixation on intervertebral biomechanics at treated and adjacent canine cervical vertebral motion units. AB - OBJECTIVE: To investigate the biomechanics of cervical vertebral motion units (VMUs) before and after a ventral slot procedure and after subsequent pin-poly methylmethacrylate (pin-PMMA) fixation and to assess the use of smooth and positive-profile threaded (PPT) pins in pin-PMMA fixation and intravertebral pin placement. SAMPLE POPULATION: Cervical portions (C3 through C6 vertebrae) of 14 cadaveric canine vertebral columns. PROCEDURE: Flexion and extension bending moments were applied to specimens before and after creation of a ventral slot across the C4-C5 intervertebral space and after subsequent smooth or PPT pin-PMMA fixation at that site. Data for the C3-C4, C4-C5, and C5-C6 VMUs were compared among treatments and between pin types, and pin protrusion was compared between pin types. RESULTS: Compared with values in intact specimens, ventral slot treatment increased neutral zone range of motion (NZ-ROM) by 98% at the treated VMUs and appeared to decrease overall ROM at adjacent VMUs; pin-PMMA fixation decreased NZ-ROM by 92% at the treated VMUs and increased overall NZ-ROM by 19% to 24% at adjacentVMUs. Specimens fixed with PPT pins were 82% (flexion) and 80% (extension) stiffer than smooth-pin-fixed specimens. Overall, 41% of pins protruded into foramina; PPT pins were more likely to protrude into transverse foramina. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that fixation of a cervical VMU alters the biomechanics of adjacent VMUs and may contribute to intervertebral degeneration of adjacent intervertebral disks. Use of threaded pins may lower the incidence of pin loosening and implant failure but enhances the likelihood of transverse foramina penetration. PMID- 15900951 TI - Seroprevalence of antibodies against Bartonella species and evaluation of risk factors and clinical signs associated with seropositivity in dogs. AB - OBJECTIVE: To determine the seroprevalence of antibodies against Bartonella spp in a population of sick dogs from northern California and identify potential risk factors and clinical signs associated with seropositivity. SAMPLE POPULATION: Sera from 3,417 dogs. PROCEDURE: Via an ELISA, sera were analyzed for antibodies against Bartonella vinsonii subsp berkhoffii, Bartonella clarridgeiae, and Bartonella henselae; test results were used to classify dogs as seropositive (mean optical density value > or = 0.350 for B henselae or > or = 0.300 for B clarridgeiae or B vinsonii subsp berkhoffi) or seronegative. Overall, 305 dogs (102 seropositive and 203 seronegative dogs) were included in a matched case control study. RESULTS: 102 of 3,417 (2.99%) dogs were seropositive for > or = 1 species of Bartonella. Of these, 36 (35.3%) had antibodies against B henselae only, 34 (33.3%) had antibodies against B clarridgeiae only, 2 (2.0%) had antibodies against B vinsonii subsp berkhoffii only, and 30 (29.4%) had antibodies against a combination of those antigens. Compared with seronegative dogs, seropositive dogs were more likely to be herding dogs and to be female, whereas toy dogs were less likely to be seropositive. Seropositive dogs were also more likely to be lame or have arthritis-related lameness, nasal discharge or epistaxis, or splenomegaly. CONCLUSIONS AND CLINICAL RELEVANCE: Only a small percentage of dogs from which serum samples were obtained had antibodies against Bartonella spp. Breed appeared to be an important risk factor for seropositivity. Bartonella infection should be considered in dogs with clinical signs of lameness, arthritis-related lameness, nasal discharge or epistaxis, or splenomegaly. PMID- 15900952 TI - Evaluation of a selective neurectomy model for low urethral pressure incontinence in female dogs. AB - OBJECTIVE: To develop a model of low urethral pressure incontinence and compare the relative contributions of the pudendal and hypogastric nerves with urethral function by performing selective neurectomy and ovariohysterectomy in dogs. ANIMALS: 19 healthy Foxhounds. PROCEDURE: Dogs were allocated into 2 groups. The first group (10 dogs) underwent bilateral hypogastric neurectomy and ovariohysterectomy and subsequent bilateral pudendal neurectomy. The second group (9 dogs) underwent bilateral pudendal neurectomy and subsequent hypogastric neurectomy and ovariohysterectomy. Urethral pressure profilometry and leak point pressure (LPP) tests were performed before and after each neurectomy. RESULTS: Before surgery, mean +/- SD LPP and maximal urethral closure pressure (MUCP) in all dogs were 169.3 +/- 24.9 cm H2O and 108.3 +/- 19.3 cm H2O, respectively; these values decreased to 92.3 +/- 27 cm H2O and 60.7 +/- 20.0 cm H2O, respectively, after both selective neurectomy surgeries. There was a progressive decline of LPP after each neurectomy; however, MUCP decreased only after pudendal neurectomy. Fifteen dogs had mild clinical signs of urinary incontinence. All dogs appeared to have normal bladder function as indicated by posturing to void and consciously voiding a full stream of urine. Urinary tract infection did not develop in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: Hypogastric and pudendal neurectomy and ovariohysterectomy caused a maximum decrease in LPP whereas pudendal neurectomy caused a maximum decrease in MUCP. IMPACT ON HUMAN MEDICINE: This model may be useful for evaluation of treatments for improving urinary control in postmenopausal women. PMID- 15900953 TI - Development of in vitro assays for the evaluation of cyclooxygenase inhibitors and predicting selectivity of nonsteroidal anti-inflammatory drugs in cats. AB - OBJECTIVE: To develop and validate in cats suitable in vitro assays for screening and ranking nonsteroidal antiinflammatory drugs (NSAIDs) on the basis of their inhibitory potencies for cyclooxygenase (COX)-1 and COX-2. ANIMALS: 10 cats. PROCEDURE: COX-1 and COX-2 activities in heparinized whole blood samples were induced with calcium ionophore and lipopolysaccharide, respectively. For the COX 2 assay, blood was pretreated with aspirin. The COX-1 and COX-2 assays were standardized, such that time courses of incubation with the test compounds and conditions of COX expression were as similar as possible in the 2 assays. Inhibition of thromboxane B2 production, measured by use of a radioimmunoassay, was taken as a marker of COX-1 and COX-2 activities. These assays were used to test 10 to 12 concentrations of a COX-1 selective drug (SC-560) and of 2 NSAIDs currently used in feline practice, meloxicam and carprofen. Selectivities of these drugs were compared by use of classic 50% and 80% inhibitory concentration (ie, IC50 and IC80) ratios but also with alternative indices that are more clinically relevant. RESULTS: These assay conditions provide a convenient and robust method for the determination of NSAID selectivity. The S(+) enantiomeric form of carprofen was found to be COX-2 selective in cats, but meloxicam was only slightly preferential for this isoenzyme. CONCLUSIONS AND CLINICAL RELEVANCE: In vitro pharmacodynamic and in vivo pharmacokinetic data predict that the COX-2 selectivity of both drugs for cats will be limited when used at the recommended doses. This study provides new approaches to the selection of COX inhibitors for subsequent clinical testing. PMID- 15900954 TI - Localization, morphology, and immunohistochemistry of spinal cord and dorsal root ganglion neurons that innervate the gastrocnemius and superficial digital flexor muscles in cattle. AB - OBJECTIVE: To determine the location, morphology, and neurochemical code of spinal cord and dorsal root ganglion neurons that innervate the gastrocnemius muscle (GM) and superficial digital flexor muscle (FDSM) in cattle. ANIMALS: 5 healthy Friesian male calves. PROCEDURE: 2 different types of neuronalretrograde fluorescent tracers (fast blue and diamidino yellow) were injected into the GM and FDSM, respectively. The neurochemical code (substance P, calcitonin gene related peptide, galanin, and neuronal nuclear protein) of labeled neurons was investigated by immunohistochemistry. RESULTS: Neurons innervating the GM and FDSM were located along the L6-S2 spinal cord segments and ganglionic levels. A cranial-to-caudal topographic distribution for each muscle was found, indicating that the motor nuclei of the 2 muscles are organized by a somatotopic pattern. The GM and FDSM motoneurons were immunoreactive only for calcitonin gene-related peptide, whereas the afferent neurons were immunoreactive for all of the neurochemical markers considered. CONCLUSION AND CLINICAL RELEVANCE: In our study, location and the extent of neurons that supply the GM and FDSM of cattle were characterized completely. Because the GM and FDSM are involved in spastic paresis of calves and it is thought that spastic paresis results from an excessive activity of the neuromuscular spindle reflex arc, findings in our study may be useful for further electrophysiologic and clinical studies. Knowledge of the neurochemical code of neurons that supply the GM and FDSM in healthy calves could be used to compare chemical alterations in the same neuronal population of affected calves. PMID- 15900955 TI - A critical role of interleukin-10 in the response of bovine macrophages to infection by Mycobacterium avium subsp paratuberculosis. AB - OBJECTIVES: To evaluate the role of interleukin (IL)-10 in the inability of monocyte-derived bovine macrophages to kill Mycobacterium avium subsp paratuberculosis organisms in vitro. SAMPLE POPULATION: Monocytes were obtained from healthy adult Holstein dairy cows that had negative results when tested for infection with M avium subsp paratuberculosis. PROCEDURE: Monocyte-derived macrophages were incubated with M avium subsp paratuberculosis for 2, 6, 24, 72, or 96 hours with or without addition of saturating concentrations of a goat anti human IL-10 that has been documented to neutralize bovine IL-10 activity. Variables assessed included ingestion and killing of M avium subsp paratuberculosis; expression of tumor necrosis factor (TNF)-alpha, IL-12, IL-8, major histocompatability (MHC) class II, vacuolar H+ ATPase, and B cell CLL/lymphoma 2 (BCL-2); production of nitric oxide; acidification of phagosomes; and apoptosis of macrophages. RESULTS: Neutralization of IL-10 enabled macrophages to kill 57% of M avium subsp paratuberculosis organisms within 96 hours. It also resulted in an increase in expression of TNF-alpha, IL-12, IL-8, MHC class II, and vacuolar H+ ATPase; decrease in expression of BCL-2; increase in acidification of phagosomes; apoptosis of macrophages; and production of nitric oxide. CONCLUSIONS AND CLINICAL RELEVANCE: The capacity of M avium subsp paratuberculosis to induce IL-10 expression may be a major determinant of virulence for this organism. PMID- 15900956 TI - Growth factors and their potential therapeutic applications for healing of musculoskeletal and other connective tissues. PMID- 15900957 TI - Potential implications of lactoferrin as a therapeutic agent. PMID- 15900958 TI - Estimating the impact of obesity. PMID- 15900959 TI - Origins and early development of the case-control study: Part 1--Early evolution. PMID- 15900960 TI - Obesity and its comorbidities: present and future importance on health status in Switzerland. AB - OBJECTIVES: The objective of this study was to estimate the proportion of various diseases attributable to obesity in Switzerland in order to get a hint of its present and future importance on health status and on our health care budgets. METHODS: The population attributable risk (PAR) for each of 17 obesity-linked conditions was estimated as the proportion of each disease condition which is attributable to obesity. The fraction of each disease that is attributable to obesity in Switzerland was calculated using the proportion (prevalence) of obesity in Switzerland and the relative risk of suffering from a given obesity related disease in Switzerland or comparable countries. RESULTS: With a PAR of 88.6% diabetes represents the disease with the highest proportion attributable to obesity in Switzerland. It is followed by a PAF of 26.8% for hypertension, 24.7% for oesophageal cancer and 24.4% for gallstones. PARs of 17.4% and 5.7% were estimated for coronary heart disease and depression. CONCLUSION: Treatment of these first four diseases represents 89% of the total health care costs attributable to obesity in Switzerland. The impact the present obesity epidemic on health status as well as its social and economic consequences must be recognised. PMID- 15900961 TI - Economic burden of obesity and its comorbidities in Switzerland. AB - OBJECTIVES: To estimate the total annual economic costs caused by overweight and obesity in Switzerland. METHODS: Top-down estimation of direct treatment costs for obesity including medication, nutritional counselling, and surgical interventions was carried out. Using Swiss prevalence data (2002) and literature based estimates of the relative risks the population attributable fraction (PAF) was calculated for 18 overweight- and obesity-related diseases. PAF was then used in combination with estimates of the total (direct and indirect) health care costs of these diseases to estimate the economic burden of obesity for Switzerland. RESULTS: The estimated total costs in Switzerland amounted to CHF 2691 million per year (on cost basis 2001). Allowance for uncertainty in parameter assumptions and values in the published literature used applying a sensitivity range of +/- 20% gave a cost range of between CHF 2153 and 3 229 millions, representing approx. 2.3-3.5% of total health care expenditures. Overweight and obesity contribute each approx. 50% to these costs. CONCLUSION: Overweight and obesity represent a considerable financial burden to the Swiss society. According to their present trends, this economic burden is expected to grow over the coming years. PMID- 15900962 TI - [Attendance at back care programmes: a representative study of the working population in Germany]. AB - OBJECTIVES: To investigate nationwide participation rates in back care programmes of the German working population. METHODS: From a national health survey conducted in Germany between October 1997 to March 1999 (n = 3313 employees aged 18 to 69), we investigated how participation in back care programmes correlates with sociodemographic characteristics, occupation, lifestyle and health factors. RESULTS: Approximately one out of six (17.5%) employee has ever attended a back care class. 7.2% took part in this kind of health promotion programme during the past year. After adjusting for various symptom prevalences, the survey revealed that males, full-time employees, singles and people with an unfavourable lifestyle were less likely to attend such programmes. CONCLUSIONS: This paper is the first to provide representative data on uptake of back care classes and investigate the correlates of participation in back care programs in Germany. The study indicates that people with the highest risk for low back pain are the least likely to take back care classes. PMID- 15900963 TI - [Differences between Eastern European immigrants of German origin and the rest of the German population in health status, health care use and health behaviour: a comparative study using data from the KORA-Survey 2000]. AB - Differences between Eastern European immigrants of German origin and the rest of the German population in health status, health care use and health behaviour: a comparative study using data from the KORA-Survey 2000 OBJECTIVES: To identify differences in health status, health care use and health behaviour between Eastern European immigrants of German origin and the rest of the German population in order to develop new concepts for preventive programs. METHODS: Using data from the KORA-Survey 2000 (Cooperative Health Research in the Region of Augsburg), immigrants of German origin were compared to the rest of the German population with multiple regression models controlling for sex, age, living with partner, years of education and occupation. Tests of trend were performed for the groups classified according to the year of immigration. RESULTS: Compared to other Germans, immigrants of German origin consider their health status more often as poor. They were more likely to be obese (OR = 1.95) and have hyperlipidemia (total-cholesterol/HDL-C > 4: OR = 1.71). They were less likely to use cancer screening (OR = 0.41) or to perform sport activities (OR = 0.47). All these differences decrease with the length of residence in Germany. CONCLUSIONS: Concerning their health status, Eastern European immigrants of German origin were identified as a high risk group. They should be considered for specific preventive programs and health care interventions, especially during the first years of immigration. PMID- 15900964 TI - Adolescent substance abuse, the importance of family, school and peers: data from the health behaviour in school children. AB - OBJECTIVES: To determine the possible factors associated with adolescent substance use in a country having recently experienced war. METHODS: The survey applied the World Health Organization research protocol for cross-national survey -HBSC: a cross-sectional study among 1540 15-year old adolescents in Belgrade was conducted. The research instrument was a self-administered questionnaire. RESULTS: Substance abuse is associated with living in a single parent family, living in a family without support, having a poor commitment to school, displaying aggressive behaviour, and spending a lot of time with friends. CONCLUSIONS: Factors associated with adolescent substance use include social background, family, school and peers. Such associations could help to focus on better measures to eliminate or reduce the risk of substance use. PMID- 15900965 TI - Unity in organisation and regulation of catabolic operons in Lactobacillus plantarum, Lactococcus lactis and Listeria monocytogenes. AB - Global regulatory circuits together with more specific local regulators play a notable role when cells are adapting to environmental changes. Lactococcus lactis is a lactic acid bacterium abundant in nature fermenting most mono- and disaccharides. Comparative genomics analysis of the operons encoding the proteins and enzymes crucial for catabolism of lactose, maltose and threhalose revealed an obvious unity in operon organisation . The local regulator of each operon was located in a divergent transcriptional direction to the rest of the operon including the transport protein-encoding genes. Furthermore, in all three operons a catabolite responsive element (CRE) site was detected inbetween the gene encoding the local regulator and one of the genes encoding a sugar transport protein. It is evident that regardless of type of transport system and catabolic enzymes acting upon lactose, maltose and trehalose, respectively, Lc. lactis shows unity in both operon organisation and regulation of these catabolic operons. This knowledge was further extended to other catabolic operons in Lc. lactis and the two related bacteria Lactobacillus plantarum and Listeria monocytogenes. Thirty-nine catabolic operons responsible for degradation of sugars and sugar alcohols in Lc. lactis, Lb. plantarum and L. monocytogenes were investigated and the majority of those possessed the same organisation as the lactose, maltose and trehalose operons of Lc. lactis. Though, the frequency of CRE sites and their location varied among the bacteria. Both Lc. lactis and Lb. plantarum showed CRE sites in direct proximity to genes coding for proteins responsible for sugar uptake. However, in L. monocytogenes CRE sites were not frequently found and not in the vicinity of genes encoding transport proteins, suggesting a more local mode of regulation of the catabolic operons found and/or the use of inducer control in this bacterium. PMID- 15900966 TI - Reassignment of enterobacter dissolvens to Enterobacter cloacae as E. cloacae subspecies dissolvens comb. nov. and emended description of Enterobacter asburiae and Enterobacter kobei. AB - The taxonomic position of Enterobacter dissolvens was re-evaluated based on the analysis of the type strain ATCC 23373T and three clinical isolates. The strains were assigned to the genetic cluster of the species by phylogenetic sequence analysis in the frame of a recent population genetic study. The relatedness of E. dissolves to the other species of the E. cloacae complex was analyzed by DNA-DNA hybridization studies based on melting profiles in microplates. The genetic cluster of E. dissolvens fell into the same DNA-relatedness group like E. cloacae with mean deltaTm-values of 3.9 degrees C confirming the hybridization results of three former studies. Phenotypic analysis of the E. cloacae and E. dissolvens strains, respectively, based on 115 biochemical reactions yielded the esculin test as the only one differentiating between them by being positive for E. dissolvens and negative for E. cloacae strains. The name E. cloacae subsp. dissolvens comb. nov. is proposed for the group of organisms formerly referred to as E. dissolvens, and the name E. cloacae subsp. cloacae comb. nov. for the group of organisms formerly referred to as E. cloacae. The species descriptions of Enterobacter kobei and Enterobacter asburiae were emended based on the data collected on 17 and 15 strains, respectively. The strains were assigned to the respective species by a combination of phylogenetic sequence analyzes and DNA-DNA hybridizations. Phenotypic analyzes of 115 reactions gave detailed species profiles with new differentiating phenotypic properties. PMID- 15900967 TI - Description of Enterobacter ludwigii sp. nov., a novel Enterobacter species of clinical relevance. AB - A new species, Enterobacter ludwigii, is presented on the basis of the characteristics of 16 strains, which were isolated from clinical specimens. These bacteria form a distinct genetic cluster in phylogenetic analyses of the population structure of the Enterobacter cloacae complex. As determined by DNA DNA cross-hybridization experiments in microplates, this genetic cluster can be delineated from the other species of the E. cloacae complex with deltaTm values equal to or above 5 degrees C with Enterobacter hormaechei being the closest relative. The bacteria are gram-negative, fermentative, motile rods with the general characteristics of the genus Enterobacter and the E. cloacae complex in particular. E. ludwigii can be differentiated from the other Enterobacter species by its growth on myo-inositol and 3-0-methyl-D-glucopyranose. The type strain is EN-119 (= DSM 16688T = CIP 108491T). PMID- 15900968 TI - Enterobacter radicincitans sp. nov., a plant growth promoting species of the family Enterobacteriaceae. AB - A plant growth promoting bacterial isolate (D5/23T) from the phyllosphere of winter wheat, able to fix atmospheric nitrogen and to produce auxines and cytokinins was investigated in a polyphasic taxonomy approach. Phylogenetic analyses using the 16S rRNA gene sequence of the strain clearly indicated that the strain belonged to the family Enterobacteriaceae, most closely related to Enterobacter cloacae with 99.0% and Enterobacter dissolvens with 98.5% sequence similarity. Phylogenetic analysis derived from the sequence of the rpoB gene showed the highest sequence similarities to Enterobacter cowanii (93.0%) but supported the distinct position of strain D5/23T. The isolate produced a fatty acid pattern typical for members of the family Enterobacteriaceae. On the basis of the phylogenetic analyses, DNA-DNA hybridizations, and the unique physiological and biochemical characteristics, we propose that strain D5/23T represents a new species of the genus Enterobacter for which we propose the name Enterobacter radicincitans sp. nov. PMID- 15900969 TI - Polymorphism of Aeromonas spp. tRNA intergenic spacers. AB - We investigated the length polymorphism of the intergenic spacers lying between tRNA genes of Aeromonas spp. A total of 69 strains representing all known genomic species of Aeromonas were used in the study. tDNA-PCR patterns were examined by Dice coefficient (S(D)) and unweighted pair group method of clustering (UPGMA). The strains were allocated into 15 groups at a similarity level of 70%. The strains belonging to seven genomic species: A. hydrophila (HG 1), A. caviae (HG 4), A. sobria (HG 7), A. veronii (HG 8/10), A. encheleia (HG 16), A. popoffii (HG 17), and A. culicicola (HG 18) formed distinct clusters. Our study revealed a genetic heterogeneity of the following species: A. bestiarum, A. salmonicida, A. media, A. eucrenophila, A. jandaei, A. schubertii, and A. allosaccharophila. PMID- 15900970 TI - Isolation and characterization of heterotrophic bacteria able to grow aerobically with quaternary ammonium alcohols as sole source of carbon and nitrogen. AB - The quaternary ammonium alcohols (QAAs) 2,3-dihydroxypropyl-trimethyl-ammonium (TM), dimethyl-diethanol-ammonium (DM) and methyl-triethanol-ammonium (MM) are hydrolysis products of their parent esterquat surfactants, which are widely used as softeners in fabric care. We isolated several bacteria growing with QAAs as the sole source of carbon and nitrogen. The strains were compared with a previously isolated TM-degrading bacterium, which was identified as a representative of the species Pseudomonas putida (Syst. Appl. Microbiol. 24 (2001) 252). Two bacteria were isolated with DM, referred to as strains DM 1 and DM 2, respectively. Based on 16S-rDNA analysis, they provided 97% (DM 1) and 98% (DM 2) identities to the closest related strain Zoogloea ramigera Itzigsohn 1868AL. Both strains were long, slim, motile rods but only DM 1 showed the floc forming activity, which is typical for representatives of the genus Zoogloea. Using MM we isolated a Gram-negative, non-motile rod referred to as strain MM 1. The 16S-rDNA sequence of the isolated bacterium revealed 94% identities (best match) to Rhodobacter sphaeroides only. The strains MM 1 and DM 1 exclusively grew with the QAA which was used for their isolation. DM 2 was also utilizing TM as sole source of carbon and nitrogen. However, all of the isolated bacteria were growing with the natural and structurally related compound choline. PMID- 15900971 TI - A molecular systematic survey of cultured microbial associates of deep-water marine invertebrates. AB - A taxonomic survey was conducted to determine the microbial diversity held within the Harbor Branch Oceanographic Marine Microbial Culture Collection (HBMMCC). The collection consists of approximately 17,000 microbial isolates, with 11,000 from a depth of greater than 150 ft seawater. A total of 2273 heterotrophic bacterial isolates were inventoried using the DNA fingerprinting technique amplified rDNA restriction analysis on approximately 750-800 base pairs (bp) encompassing hypervariable regions in the 5' portion of the small subunit (SSU) 16S rRNA gene. Restriction fragment length polymorphism patterns obtained from restriction digests with RsaI, HaeIII, and HhaI were used to infer taxonomic similarity. SSU 16S rDNA fragments were sequenced from a total of 356 isolates for more definitive taxonomic analysis. Sequence results show that this subset of the HBMMCC contains 224 different phylotypes from six major bacterial clades (Proteobacteria (Alpha, Beta, Gamma), Cytophaga, Flavobacteria, and Bacteroides (CFB), Gram + high GC content, Gram + low GC content). The 2273 microorganisms surveyed encompass 834 alpha-Proteobacteria (representing 60 different phylotypes), 25 beta-Proteobacteria (3 phylotypes), 767 gamma-Proteobacteria (77 phylotypes), 122 CFB (17 phylotypes), 327 Gram + high GC content (43 phylotypes), and 198 Gram + low GC content isolates (24 phylotypes). Notably, 11 phylotypes were < or =93% similar to the closest sequence match in the GenBank database even after sequencing a larger portion of the 16S rRNA gene (approximately 1400 bp), indicating the likely discovery of novel microbial taxa. Furthermore, previously reported "uncultured" microbes, such as sponge-specific isolates, are part of the HBMMCC. The results of this research will be available online as a searchable taxonomic database (www.hboi.edu/dbmr/dbmr_hbmmd.html). PMID- 15900972 TI - Diverse bacteria isolated from root nodules of Phaseolus vulgaris and species within the genera Campylotropis and Cassia grown in China. AB - Eighty bacterial isolates from root nodules of the leguminous plants Phaseolus vulgaris, Campylotropis spp. and Cassia spp. grown in China were classified into five groups by phenotypic analyses, SDS-PAGE of whole-cell proteins, PCR-based 16S rRNA gene restriction-fragment-length-polymorphism and sequencing. Thirty three isolates from the three plant genera were identified as Agrobacterium tumefaciens because they are closely related to the type strain of A. tumefaciens. Fourteen isolates from P. vulgaris grown in Yunnan and Inner Mongolia were classified as R. leguminosarum bv. phaseoli based on their close relationship with the type strain in numerical taxonomy and in 16S rDNA phylogeny. Twenty-seven isolates from Campylotropis delavayi, P. vulgaris and four species of Cassia grown in the central zones of China were classified into three groups within the genus Bradyrhizobium. One of these three groups could be defined as Bradyrhizobium japonicum. Our results demonstrated that P. vulgaris and the species of Campylotropis and Cassia could form nodules with diverse rhizobia in Chinese soils, including novel lineages associated with P. vulgaris. These results also offered information about the convergent evolution between rhizobia and legumes since the rhizobial populations associated with P. vulgaris in Chinese soils were completely different from those in Mexico, the original cite of this plant. Some rhizobial species could be found in all of the three leguminous genera. PMID- 15900973 TI - Natural association of Gluconacetobacter diazotrophicus and diazotrophic Acetobacter peroxydans with wetland rice. AB - The family Acetobacteraceae currently includes three known nitrogen-fixing species, Gluconacetobacter diazotrophicus, G. johannae and G. azotocaptans. In the present study, acetic acid-producing nitrogen-fixing bacteria were isolated from four different wetland rice varieties cultivated in the state of Tamilnadu, India. Most of these isolates were identified as G. diazotrophicus on the basis of their phenotypic characteristics and PCR assays using specific primers for that species. Based on 16S rDNA partial sequence analysis and DNA: DNA reassociation experiments the remaining isolates were identified as Acetobacter peroxydans, another species of the Acetobacteraceae family, thus far never reported as diazotrophic. The presence of nifH genes in A. peroxydans was confirmed by PCR amplification with nifH specific primers. Scope for the findings: This is the first report of the occurrence and association of N2-fixing Gluconacetobacter diazotrophicus and Acetobacter peroxydans with wetland rice varieties. This is the first report of diazotrophic nature of A. peroxydans. PMID- 15900974 TI - My time. PMID- 15900975 TI - Now you see and now you don't. PMID- 15900976 TI - Laparoscopic radical nephrectomy in renal cell carcinoma. AB - Historically, the only surgical option for removal of a renal mass was through a large incision in the abdomen. Recently, many urologists have adopted new innovations in the removal of renal masses including laparoscopic radical nephrectomy. The laparoscopic approach to radical nephrectomy has been associated with improved patient outcomes including decreased pain, shorter hospital stay, rapid recovery, and improved aesthetic cosmetic appearance. Laparoscopic radical nephrectomy may be performed in three different types of surgical procedures including laparoscopic hand-assisted radical nephrectomy, transperitoneal radical nephrectomy, and retroperitoneal radical nephrectomy. Understanding the procedure, preoperative instructions, and recovery will assist nurses in counseling patients considering laparoscopic radical nephrectomy. PMID- 15900977 TI - Individual quality of life following radical prostatectomy in men with prostate cancer. AB - PURPOSE: The aim of this study was to examine the individual quality of life (QoL) of men following radical prostatectomy for prostate cancer. The following research questions were addressed: (a) What are the most important areas of quality of life for men following radical prostatectomy? (b) How do these men rate their satisfaction in each area and what is the relative importance of each area to their overall quality of life? METHODS: The purposive sample consisted of 11 men with prostate cancer who had undergone a radical prostatectomy 3 to 4 months earlier. QoL was examined using the SEIQoL-DW (Schedule for the Evaluation of Individual QoL: A Direct Weighting Procedure). The data were analyzed by means of qualitative content analysis (five most important QoL areas). FINDINGS: The 11 respondents named a total of 55 QoL areas which they described and labelled. They then rated their current satisfaction in each area, and how important each one was to them. A second analysis of the content was made to identify the main QoL areas. The 55 quality of life areas mentioned by respondents were reduced to the following categories: health, activity, family, relationship with a partner, autonomy, independence, hobby, financial security, and sexuality. Health, family, and relationship with a partner are the thee areas which had the most impact on QoL. Overall, the respondents had a high quality of life value. Impotence and incontinence did not appear to have a very negative impact on quality of life. CONCLUSIONS: SEIQoL-DW was used for the first time in patients with prostate cancer. In a urology department where nurses and patients are confronted daily with the topics of intimacy, sexuality, and sense of embarrassment, more importance should be placed on the topic of sexuality when taking a patient history. Nurses should be trained in communication techniques that enable them to engage patients in a safe and therapeutic dialogue about their sexual concerns related to the diagnosis of prostate cancer. SEIQoL-DW can support the communication with patients. PMID- 15900978 TI - Use of a continence nurse specialist in an extended care facility. AB - PURPOSE: The aim of this study was to determine if the number of incontinence episodes for an elderly female population could be decreased through an individualized continence program in a Connecticut long-term care center. METHODS: Forty-two female residents who in a long-term care facility who were incontinent or had urgency related to overactive bladder were included in the incontinence program. Total numbers of incontinent episodes for each participant were recorded 1 week prior to the study. An individualized plan of care for each patient was developed by the continence specialist and the plan of care implemented for at least 1 year. FINDINGS: After the continence specialist recommended a program of treatment and the program of treatment was implemented for 1 year, the number of UTIs (31 preintervention year; 6 postintervention year) and pressure ulcers (15 preintervention year; 2 postintervention year) were substantially decreased and the number of falls cut by more than 50% (18 preintervention; 7 postintervention). CONCLUSION: A nurse continence specialist can be used to help long-term care facilities plan a program that will direct individualized nursing interventions that will improve patient outcomes related to UTI, pressure sore and fall rates, and reduce the costs of care. PMID- 15900979 TI - What Americans understand and how they are affected by bladder control problems: highlights of recent nationwide consumer research. AB - Over the last 5 years, the National Association For Continence has sponsored epidemiologic surveys of adults across the United States in order to quantify the prevalence of bladder control problems in the community, to understand how quality of life of those people experiencing these medical disorders is affected, and what beliefs people in general hold surrounding bladder health. Collectively, these nationwide surveys offer statistically significant documentation of how widespread, how far-reaching, how misunderstood, and how real the symptoms of urinary incontinence and related symptoms of overactive bladder are, and lay a foundation to support future investment in research, education, and collaboration. PMID- 15900980 TI - Exertional rhabdomyolysis in unsupervised exercises in a correctional setting: a case study. AB - Unsupervised overexercise in the prison population can lead to exertional rhabdomyolysis, a condition that can lead to kidney damage and/or failure. Knowledge deficit of the risk from this activity will be addressed. Signs and symptoms of rhabdomyolysis and its sequelae will be presented along with a case study. Inter-departmental responsibilities in the correctional setting will be considered. The importance of recognizing the signs and symptoms of this condition by nurses in various clinical settings will also be explored. PMID- 15900981 TI - A bond of trust: delegation. PMID- 15900982 TI - The interview in qualitative research. PMID- 15900983 TI - Update on foodborne diseases. PMID- 15900985 TI - Getting ready for certification: urinary calculi. PMID- 15900986 TI - Abandoning isolated free PSA screenings for more comprehensive cardiovascular and cancer screening and education. AB - The time is ripe to abandon free PSA screenings and replace them with free comprehensive health screenings. For example, cardiovascular disease (CVD) risk screening (cholesterol, blood pressure) that could also include PSA, weight measurement, and a variety of other tests and educational assessments just make more sense in terms of the larger picture of influencing all-cause morbidity and mortality. Numerous reasons are proffered for this important change. Abandoning this myopic single-disease only approach must be embraced by numerous medical centers and health professionals before this comprehensive approach can be successfully implemented. PMID- 15900987 TI - Guillain-Barre--not an Irish beer. PMID- 15900988 TI - Weight changes, exercise, and dietary patterns during freshman and sophomore years of college. AB - Weight gain and behavioral patterns during college may contribute to overweight and obesity in adulthood. The aims of this study were to assess weight, exercise, and dietary patterns of 764 college students (53% women, 47% men) during freshman and sophomore years. Students had their weight and height measured and completed questionnaires about their recent exercise and dietary patterns. At the beginning of freshman year, 29% of students reported not exercising, 70% ate fewer than 5 fruits and vegetables daily, and more than 50% ate fried or high-fat fast foods at least 3 times during the previous week. By the end of their sophomore year, 70% of the 290 students who were reassessed had gained weight (4.1+/-3.6 kg, p < .001), but there was no apparent association with exercise or dietary patterns. Future research is needed to assess the contributions of fat, muscle, and bone mass to observed weight gain and to determine the health implications of these findings. PMID- 15900989 TI - Prevalence and motives for illicit use of prescription stimulants in an undergraduate student sample. AB - To assess the prevalence and motives for illicit use of prescription stimulants and alcohol and other drugs (AODs), associated with these motives, the authors distributed a self-administered Web survey TO a random sample of 9,161 undergraduate college students. Of the study participants, 8.1% reported lifetime and 5.4% reported past-year illicit use of prescription stimulants. The most prevalent motives given for illicit use of prescription stimulants were to (1) help with concentration, (2) increase alertness, and (3) provide a high. Although men were more likely than women were to report illicit use of prescription stimulants, the authors found no gender differences in motives. Regardless of motive, illicit use of prescription stimulants was associated with elevated rates of AOD use, and number of motives endorsed and AOD use were positively related. Students appear to be using these prescription drugs non-medically, mainly to enhance performance or get high. PMID- 15900990 TI - My student body: a high-risk drinking prevention web site for college students. AB - The authors investigated the efficacy of an interactive Web site, MyStudentBody.com: Alcohol (MSB:Alcohol) that offers a brief, tailored intervention to help heavy drinking college students reduce their alcohol use. They conducted a randomized, controlled clinical trial to compare the intervention with an alcohol education Web site at baseline, postintervention, and 3-month follow-up. Students were assessed on various drinking measures and their readiness to change their drinking habits. The intervention was especially effective for women and persistent binge drinkers. Compared with women who used the control Web site, women who used the intervention significantly reduced their peak and total consumption during special occasions and also reported significantly fewer negative consequences related to drinking. In addition, persistent heavy binge drinkers in the experimental group experienced a more rapid decrease in average consumption and peak consumption compared with those in the control group. The authors judged MSB:Alcohol a useful intervention for reaching important subgroups of college binge drinkers. PMID- 15900991 TI - Social support and physical health: the importance of belonging. AB - Social support is a multifaceted construct recognized as a significant predictor of physical health. In this study, the authors examined several support domains simultaneously in a sample of 247 college students to determine their unique prediction of physical health perceptions and physical symptoms. They also examined gender differences across social support and physical health. Their findings indicated that belonging predicted better health perceptions for women and fewer physical symptoms for men, suggesting that a sense of connection to a group of others is a key support component for the physical health of college students. PMID- 15900992 TI - Relationships between physical activity and the proximity of exercise facilities and home exercise equipment used by undergraduate university students. AB - The authors used stratified random sampling procedures to investigate the relationships among physical activity (PA), the proximity of exercise facilities, and the quantity of home exercise equipment in a sample of 411 undergraduates. To examine the data they collected from the modified Godin Leisure-Time Exercise Questionnaire and the Home Environment Questionnaire, the authors used correlational analyses and analysis of variance. Intensity and duration of PA showed a significant relationship to the proximity of exercise facilities. Intensity, frequency, and duration of PA were significantly related to the quantity of students' home exercise equipment. Freshmen and sophomores engaged in PA closer to their residences and participated in 3 more exercise bouts per week than juniors and seniors. These findings suggest that proximity of the activity episode may have an effect on PA behavior of college students. PMID- 15900993 TI - Responses of US college and university student health services to the 2004 influenza vaccine shortage. AB - The United States experienced a shortage of influenza vaccine for the 2004--2005 influenza season. The authors surveyed college health programs to determine whether they had targeted vaccine to priority groups and knew how to reallocate remaining vaccine. They used an electronic message to distribute a Web-based survey to the members of 3 college-affiliated organizations--the Association of American Colleges and Universities, American Association of Community Colleges, American College Health Association--and to subscribers of the Student Health Service Listserv. They received 434 completed surveys. Sixty percent (259) of the respondents stated they had received vaccine and planned to vaccinate their high risk students, staff, and faculty members; 77% (198) planned to reallocate leftover vaccine. Given the potential for future disruptions of the influenza vaccine supply, the authors recommend that college health programs establish policies to identify members of their high-risk population and also consider providing the live attenuated influenza virus vaccine. PMID- 15900994 TI - Academic incentives for students can increase participation in and effectiveness of a physical activity program. AB - The authors sought to determine whether a greater academic incentive would improve the effectiveness and student adherence to a 12-week voluntary exercise program designed to decrease students' percentage of body fat. They randomly assigned 210 students to 1 of 2 groups with different academic reward structures. The group with the greater reward structure showed better exercise adherence and lost more body fat than those without the additional incentive. These findings suggest that an academic incentive can increase overall student adherence to a voluntary exercise program and can boost the effectiveness of the program in a university environment. The findings also have potential implications for on campus promotion of physical activity. PMID- 15900995 TI - Survival in high-grade glioma: a study of survival in patients unfit for or declining radiotherapy. AB - AIMS: To determine the survival of patients with high-grade glioma (HGG) and a poor prognosis in terms of age or performance status managed with best supportive care alone. METHODS: An analysis of survival was carried out on 123 patients with HGG declining or judged unfit to receive radiotherapy, on the basis of age or performance status, between February 1998 and October 2003. Karnofsky performance status (KPS), biopsy or resection or no surgery, attendance at clinic and reason for not receiving radiotherapy were prospectively recorded. RESULTS: Of the 123 patients, three were excluded from the analysis, as no outcome data were available. Median age was 66 years (range 29-91 years), and median KPS was 50 (range 30-100). All 120 patients included had died at the time of analysis. Overall median survival was 68 days (95% CI 56-85), range 2-294 days and interquartile range 35-123 days. Median survival of 22 patients declining radiotherapy was 75 days (95% CI 53-123), of 98 patients unfit for radiotherapy 67 days (95% CI 48-88); non-significant difference P = 0.36. Median survival of 26 patients undergoing biopsy was 95 days (95% CI 66-123), of 56 undergoing surgical resection 74 days (95% CI 47-93), and of 38 receiving no surgical intervention 59 days (95% CI 47-70); non-significant difference P = 0.16. CONCLUSION: For patients with HGG and a poor prognosis, in terms of age or performance status managed with best supportive care, survival is short. Survival may be too short to benefit from radiotherapy and possibly surgery. PMID- 15900996 TI - Treatment of recurrent head and neck cancer: re-irradiation or chemotherapy? AB - For most patients with head and neck cancer, locoregional disease recurrence carries an extremely poor prognosis and has severe adverse effects on quality of life. Only a few patients are suitable for salvage surgery and, even in selected cases, the success rate is low. Most patients are managed by supportive palliative care, or with palliative chemotherapy. In the UK, re-irradiation is rarely used because of concerns about treatment-related toxicity and lack of efficacy. Despite this, a significant body of evidence suggests that re irradiation may have a higher probability of achieving local control than other treatments. In this review, we discuss the use of re-irradiation in patients with locally recurrent head and neck cancer, and present the pertinent data. PMID- 15900997 TI - Comparison of induction chemotherapy with docetaxel, cisplatin, and 5 fluorouracil (TPF) followed by radiation vs concurrent chemoradiotherapy with TPF in patients with locally advanced squamous cell carcinoma of the head and neck. AB - AIMS: To compare the effectiveness of induction chemotherapy with docetaxel, cisplatin and 5-fluorouracil (TPF) followed by radiation with that of concurrent chemoradiotherapy with TPF in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN). MATERIALS AND METHODS: In a group of patients receiving induction chemotherapy followed by radiation, 15 patients received two cycles of chemotherapy with docetaxel 60 mg/m2, cisplatin 70 mg/m2 and 5-day 5-fluorouracil (5-FU) 750 mg/m2/day. Radiotherapy was begun 21 days after completing chemotherapy. In the group receiving concurrent chemoradiotherapy, 19 patients received two cycles of chemotherapy with docetaxel 50 mg/m2, cisplatin 60 mg/m2, and 5-day 5-FU 600 mg/m2/day. Radiation was begun on the first day of chemotherapy. The total radiation dose was between 63 and 74 Gy. RESULTS: Overall response rate (partial and complete response--both 100%) and complete response rate (87% and 84%) were similar, but, in overall survival, concurrent chemoradiotherapy with TPF was better than induction chemotherapy with TPF followed by radiation. Mucositis and anaemia were more frequent in the group receiving concurrent chemoradiotherapy, but the group receiving concurrent chemoradiotherapy with TPF improved overall survival. CONCLUSIONS: This is a small non-randomised comparison. The effectiveness of concurrent chemoradiotherapy with TPF was better than that of induction chemotherapy with TPF followed by radiation. PMID- 15900998 TI - Cardiac complications of radiation therapy. AB - AIMS: To present an overview of cardiac complications arising from radiation therapy. MATERIALS AND METHODS: Medline the (February 2004) and Embase (1974 to February 2004) searches of the medical literature relating to the cardiac complications of radiotherapy were conducted. RESULTS: Radiation damage may affect the pericardium, myocardium or coronary vasculature, and consists of fibrotic or small vessel damage. Cardiac complications are a particular problem with radiation treatments to the mediastinum and breast, especially when greater than 65% of the heart is irradiated. Most of the literature relates to the treatment of Hodgkin's disease, as patients with this disease tend to be young and live long enough to manifest late cardiac complications. Pericarditis, angina, myocardial infarction and arrhythmias are the most frequent causes of morbidity, with myocardial infarction being the most common fatal complication. The incidence of ischaemic heart disease does not increase rapidly until 10 years from treatment. CONCLUSIONS: Much of the evidence relates to the use of outdated radiation therapy equipment and techniques. Today's patients almost certainly have a lower risk of cardiac complications. Cardiac complications are probably under-reported, as they occur long after cured patients have been discharged from follow-up. PMID- 15900999 TI - Concomitant chemoradiotherapy for muscle-invasive bladder cancer: the way forward for bladder preservation? AB - Muscle-invasive bladder cancer is a common malignancy with a high mortality rate. Despite ongoing debates about the optimal primary intervention, radical cystectomy remains the cornerstone of first-line therapy in many institutions. Over the past decade, bladder-preserving strategies involving transurethral resection (TUR), chemotherapy and radiotherapy have evolved. However, the advantage of these approaches over radiation treatment as monotherapy has yet to be fully evaluated. In other tumour models, most notably cervical and anal cancer, radiation and chemotherapy delivered concomitantly have resulted in significant survival advantages. Here, we consider the potential value of this approach in the treatment of invasive bladder cancer. Concomitant chemoradiotherapy is currently the mainstay of several bladder-preserving programmes reported in the medical literature. Overall, local control and survival rates compare favourably with contemporary cystectomy series; however, difficulties in drawing valid conclusions are highlighted. Concomitant chemoradiotherapy may have a role in the management of certain patient subgroups, and the debate should remain open. Further large-scale randomised trials are needed, and information regarding bladder function and quality of life after treatment is lacking at present. The importance of close follow-up and prompt salvage cystectomy is emphasised. PMID- 15901000 TI - Tumour staging using magnetic resonance imaging in clinically localised prostate cancer: relationship to biochemical outcome after neo-adjuvant androgen deprivation and radical radiotherapy. AB - AIMS: To evaluate the prognostic significance of magnetic resonance imaging (MRI) tumour stage in clinically localised prostate cancer. MATERIALS AND METHODS: Between 1988 and 1999, 199 men with clinically localised prostate cancer (T -T4, N0/Nx, M0) were treated with neo-adjuvant androgen deprivation and radical radiotherapy, and were staged using MRI. Concordance between clinical tumour (cT) stage, as determined by digital rectal examination, and MRI tumour (mT) stage was assessed. Univariate and multivariate analyses using the Cox proportional hazards model were used to study the prognostic role of cT stage and mT stage in addition to established prognostic factors. RESULTS: Of these 199 patients, 103 (52%) were upstaged on MRI, seven (3%) were downstaged, and in 89 (45%) cT and mT stages were concordant. With median follow-up of 3.8 years, 5-year freedom from prostate specific antigen (PSA) failure was 48% (95% confidence interval (CI) 39-56%). On univariate analysis, freedom from PSA failure was associated with mT stage (P = 0.009) as well as Gleason score (P < 0.001) and initial PSA (P < 0.001), but not cT stage (P = 0.449). On multivariate analysis, Gleason score (P = 0.001), initial PSA (P < 0.001), but not mT stage (P = 0.112) remained independent determinants of freedom from PSA failure. For the subgroup of 149 patients with cT1-2 disease, mT stage was a significant predictor of increased risk of PSA failure on univariate analysis (P = 0.005), but not multivariate analysis (P = 0.19). CONCLUSION: Freedom from PSA failure was more closely associated with mT stage than cT stage. Future studies are warranted to determine whether mT stage is an independent determinant of treatment outcome. PMID- 15901001 TI - The utility of digital rectal examination after radical radiotherapy for prostate cancer. AB - AIMS: The aim of the current study was to determine the utility of routine digital rectal examination (DRE) after radical radiotherapy for prostate cancer. MATERIALS AND METHODS: Between 1990 and 1999, 899 patients with clinically localised prostatic adenocarcinoma (T1-4, N0/Nx, M0/Mx) underwent neoadjuvant androgen deprivation and radical radiotherapy at the Royal Marsden Hospital. Patients were followed with serum prostate-specific antigen (PSA) test and DRE carried out at 6-monthly intervals for the first 2 years, and then annually. RESULTS: At a median follow-up of 5 years, 39 out of 899 cases (4.3%) had local recurrence detected on DRE. DRE failed to detect any local recurrences in the absence of a rising PSA. The lowest serum PSA concentration at the time of clinically detectable local recurrence was 1.7 ng/ml. CONCLUSIONS: These findings question the standard model of follow-up after radiotherapy for prostate cancer, and suggest that alternatives, such as telephone clinics, should be considered. PMID- 15901002 TI - What's new in the management of cutaneous T-cell lymphoma? AB - The aetiology and clinical management of primary cutaneous T-cell lymphoma (CTCL) and specifically of mycosis fungoides and Sezary syndrome are poorly defined. Interesting new insights into CTCL disease biology as well as a number of emerging of novel therapeutic interventions make this an increasingly interesting area for dermatologists and oncologists involved in the treatment of CTCL. This review article covers much of this new information including new drugs, such as denileukin diftitox (Ontak) a targeted cytotoxic biological agent, Bexarotene an RXR selective retinoid, anti-CD4 monoclonal antibodies (mAb), new cytotoxics agents and vaccines. PMID- 15901003 TI - Primary central nervous system lymphoma: a single-centre experience of 55 unselected cases. AB - AIMS: Current treatment for primary central nervous system lymphoma (PCNSL) involves high-dose methotrexate (HDMTX) with or without radiotherapy. Many published studies describing this approach include a highly selected group of patients. We report a single-centre experience of unselected cases of PCNSL. MATERIALS AND METHODS: We retrospectively reviewed the case notes of 55 consecutive patients diagnosed with biopsy-proven PCNSL between 1995 and 2003 at Addenbrooke's Hospital Cambridge, UK. We describe the treatment and outcome, including survival, treatment-related toxicity and long-term functional disability. RESULTS: At diagnosis, 45% of patients were considered unfit to receive treatment with HDMTX, owing to poor performance status or comorbidity. These patients had a median survival of 46 days and may not have been included in other published studies. The remaining patients were treated with a chemotherapy regimen, which included HDMTX. Patients who received at least one cycle of a chemotherapy containing HDMTX had a median survival of 31 months. Forty per cent did not complete planned chemotherapy owing to toxicity, disease progression or death. The median survival of patients treated with HDMTX aged 60 years compared with patients aged under 60 years was 26 months vs 41 months (P = 0.07), respectively. Younger patients treated with HDMTX, who achieved complete remission with chemotherapy, had a median survival of 56 months. We identified a high incidence of functional disability among survivors, resulting from a combination of the tumour itself, the neurosurgical procedure required for diagnosis and the late neurotoxicity of combined chemoradiotherapy. CONCLUSION: The treatment of PCNSL is associated with significant early and late toxicity. Further attempts to improve treatment should address mechanisms to reduce this toxicity. In particular, the benefit of radiotherapy in patients who achieve complete remission with HDMTX will remain uncertain until it is addressed in a multicentre, randomised trial. PMID- 15901004 TI - An audit of the indications for and techniques of palliative splenic radiotherapy in the UK. AB - AIMS: This paper describes a national audit of the indications for, and techniques and toxicity of, palliative splenic radiotherapy in haematological disorders. MATERIALS AND METHODS: A postal questionnaire was sent to consultant clinical oncologists treating haematological malignancies in the UK. RESULTS: The response rate was 76%. The audit shows chronic lymphocytic leukaemia, myelofibrosis and chronic myeloid leukaemia to be the most common underlying conditions in which splenic irradiation is used. Painful splenomegaly and hypersplenism were the most common indications. Dose fractionation schedules vary widely across the UK, and also the level of full blood counts used to interrupt radiotherapy. CONCLUSIONS: Palliative splenic radiotherapy continues to be widely used in the UK in small numbers of patients and seems to be well tolerated. Re irradiation for further symptomatic progression is also commonly carried out. PMID- 15901005 TI - Natural history of tumour-related sacral obliteration with nerve-root preservation. AB - Benign aggressive bone tumours can present a dilemma when the definitive treatment options necessitate enormous and permanent functional deficits. Here, we present a case of a massive sacral giant-cell tumour causing dramatic skeletal obliteration, which was successfully treated with radical radiotherapy rather than ablative surgery. The excellent functional outcome highlights the importance of nerve-root preservation in selecting treatment modalities. PMID- 15901006 TI - Work-based learning, role extension and skills mix within dose planning: target volume definition for carcinoma of the prostate by non-clinicians. PMID- 15901007 TI - Malignant melanoma and SIADH. PMID- 15901008 TI - Imbalanced synthesis of interleukin-1beta and interleukin-1 receptor antagonist: prognostication in intestinal-type gastric cancer. PMID- 15901009 TI - Primary vascular leiomyosarcoma of the femoral vein leading to metastases of scalp and lungs. PMID- 15901011 TI - Marketing dentistry, where to next? PMID- 15901010 TI - Primary small-cell carcinoma of the breast. PMID- 15901013 TI - The "paper chase" in medical aid claims. PMID- 15901012 TI - The frequency of culturally derived dental modification practices on the Cape Flats in the Western Cape. AB - The people living on the Cape Flats (Western Cape) have been practicing dental modification by having their incisors removed for at least 60 years. A survey, by means of a questionnaire, of eight adjoining areas in the Northern Suburbs was done to investigate the current prevalence of this practice. Three groups of study subjects (scholars, working people and retired people) were included to gain a perspective of the community in general. The individual ages ranged between 15 and 83 years old. A total of 2 167 individuals participated in this study of which 41% had modified their teeth. More males (44.8%) than females (37.9%) were involved in this practice. The percentage incidence was overwhelmingly higher in lower income areas. 'Race' and social class are both important factors. Of people who called themselves 'coloured', 74.5% modified their teeth, but 39.8% of 'blacks' and 31.6% of 'whites' within the sample had also practiced tooth removal. The frequency of tooth removal for those who identified themselves as South African was much lower (22.9%). This suggests that dental modification is a phenomenon of ethnicity and community and those who perceive themselves to be part of a broader national fabric are less likely to practice it. PMID- 15901014 TI - The bonding effectiveness of five luting resin cements to the IPS Empress 2 all ceramic system. AB - BACKGROUND: Variolink II is the only resin cement used for bonding IPS (Ivoclar Porcelain System) Empress 2 ceramic restorations. Alternative luting resin cements need to be investigated for their bonding effectiveness with the IPS Empress 2 ceramic. OBJECTIVES: To determine the shear bond strength (SBS) and the effect of thermocycling, on the bonding effectiveness, of five resin cements to IPS Empress 2 ceramic. MATERIALS AND METHODS: The projecting surfaces of one hundred ceramic discs were ground wet on silicone carbide paper. The specimens were divided into 5 groups of 20. The resin cements were bonded to the prepared ceramic surfaces, in the form of a stub. The specimens were stored under distilled water at 37 degrees C in an oven for 24 hours. Ten specimens in each group were thermocycled for 300 cycles between 5 degrees C and 55 degrees C. All the specimens were stressed to failure in an Instron Materials Testing Machine. RESULTS: The results were subjected to a one-way analysis of variance (ANOVA). Statistically similar mean SBS values were grouped using the Bonferroni (Dunn) multiple comparison test. The means for the non-thermocycled group were: 26.21, 19.41, 17.69, 17.43, and 15.76. The means for the thermocycled group were: 22.90, 15.72, 14.34, 13.96 and 13.45. The differences between the means were highly significant (p < 0.0125). CONCLUSIONS: The shear bond strength of Variolink II and Rely XARC to IPS Empress 2 ceramic was effective. Thermocycling had a significant effect on the mean SBS values of Calibra. Thermocycling had no significant effect on the mean SBS values of the other resin cements. PMID- 15901015 TI - Effect of saliva contamination on microleakage of a bonding system. AB - Saliva contamination is a major clinical problem during restorative dental treatment. Restorative systems must possess sufficient marginal sealing ability to avoid microleakage. PURPOSE: The purpose of this study was to investigate the effect of saliva contamination on the microleakage of cavities restored with Prime & Bond NT and Dyract-AP. METHOD: Cavities were prepared with occlusal margins in enamel and gingival margins in dentine on the buccal surfaces of extracted human permanent premolar teeth. The cavities were etched with phosphoric acid or a non rinse conditioner (NRC). Preparations were distributed randomly into 4 treatment groups (N = 18) consisting of two respective etching methods (phosphoric acid/NRC), each with and without saliva contamination. After the restorative process the samples were thermocycled in dye, sectioned, and scored for microleakage. RESULTS: Contamination with saliva had no significant effect on microleakage on the enamel or dentine sides where phosphoric acid was used as conditioner. Where NRC was used, significantly more microleakage was observed at the enamel sides, but significantly less at the dentine sides. PMID- 15901017 TI - Certificate of Need. PMID- 15901016 TI - Orthodontic space maintenance--a review of current concepts and methods. AB - Crowding and a lack of space in the permanent dentition are amongst the most common orthodontic problems encountered in orthodontic patients. Often the aetiology of these problems is linked to premature loss of one or more deciduous teeth with corresponding loss of space. Many of these cases could have been prevented or their severity alleviated if the practitioner employed adequate planning and space maintenance during initial treatment in the mixed dentition. A review of some of the current orthodontic space maintenance appliances and their indications is presented along with a table to aid the practitioner in treatment planning. PMID- 15901018 TI - SADA fees as a basis from which to calculate fair fees for 2005. PMID- 15901019 TI - The management of type II diabetes. AB - Type II diabetes mellitus is one of the most common chronic diseases seen by general practitioners. The seriousness of the disease and its consequences for the patient are often underestimated. Insight into the pathophysiology of the disease often helps to improve the management of these patients. It is therefore crucial to know how to treat insulin resistance and the associated metabolic syndrome. Knowledge of the different components of this dreadful syndrome is central to the success of treating these patients. PMID- 15901020 TI - The differential diagnosis of heart failure. PMID- 15901021 TI - The metabolic syndrome, insulin resistance and cardiovascular disease. AB - The metabolic syndrome (MS) is a common lifestyle disease in the western world and has significant morbidity and premature mortality, especially regarding cardiovascular disease (CVD). Although insulin resistance (and hyperinsulinaemia) is an early marker of MS and future adverse cardiovascular outcomes, it is not known if on its own this is sufficient. The issue is further clouded in prospective studies by the development in study subjects of some, or all of the components of MS, each of which is an independent risk factor for CVD. Therefore, in spite of a number of appropriate long-term studies, the exact contributions of the individual components of MS remain unclear. Their combination is unequivocally responsible for this presentday epidemic of CVD. PMID- 15901022 TI - General practitioner's radiology. Case 30. Mixed radiolucent/ radiopaque lesion in the 45-46 regions. PMID- 15901023 TI - [Designer drugs and anesthesia]. PMID- 15901024 TI - [Routine preoperative screening of asymptomatic patients: beliefs and attitudes of anesthesiologists in the Canary Islands, Spain]. AB - OBJECTIVES: To identify factors that explain anesthesiologists' inappropriate use of preoperative tests in asymptomatic patients scheduled for elective surgery. MATERIAL AND METHODS: A validated questionnaire was sent to anesthesiologists at university hospitals in the Canary Islands. Information about preoperative testing patterns and reasons for selecting tests was gathered. RESULTS: The questionnaire was self-administered by 66 anesthesiologists (68% of the total). Scientific knowledge was not the reason why most respondents ordered preoperative tests in asymptomatic individuals. That was the opinion of 95% of anesthesiologists for chest x-rays, 82% for electrocardiograms, and 68% for laboratory tests. Clinical history and a medical examination gave sufficient information for selecting patients in need of specific tests in the opinion of 77.19% of the respondents. Half did not agree that routine electrocardiograms and laboratory tests should be abandoned. The justification for these tests was coverage of medical malpractice liability for 68.42%. Most considered that the need to order preoperative tests in asymptomatic patients increased after age 40. CONCLUSIONS: Although anesthesiologists admit that their request for preoperative tests in asymptomatic individuals is not supported by scientific evidence, the quest for safety and legal protection from the consequences of potential adverse consequences of providing anesthesia affects the selection of preoperative tests for asymptomatic patients. PMID- 15901025 TI - [Prevalence and risk factors for early postoperative infection after liver transplantation]. AB - OBJECTIVES: To analyze factors related to the development of infection soon after a liver transplant. PATIENTS AND METHOD: Retrospective study of 1000 liver transplants in adults between 1991 and 2004. Pre-, intra- and postoperative variables of recipients were analyzed in 2 groups according to whether infection did or did not develop. RESULTS: Infection developed in 151 patients. Bacterial infections were the most common type. Significant risk factors for infection in the multivariate analysis were sex (odds ratio [OR], 0.54; 95% confidence interval [CI], 0.33-0.90); Child-Pugh stage (OR, 1.89; 95% CI, 1.29-2,77); hepatitis C virus cirrhosis (OR, 0.58; 95% CI, 0.34-0.99); post-reperfusion syndrome (OR, 1.82; 95% CI, 1.03-3.21); vena cava preservation technique (OR, 0.43; 95% CI, 0.22-0.84); history of diabetes mellitus (OR, 2.38; 95% CI, 1.34 4.22); respiratory distress syndrome (OR, 6.60; 95% CI, 1.16-37.45); pulmonary edema (OR, 2.36; 95% CI, 1.44-3.86); renal dysfunction (OR, 3.25; 95% CI, 1.89 5.60); acute renal insufficiency (OR, 20.24; 95% CI, 9.88-41.46); neurological alterations (OR, 3.36; 95% CI, 1.94-5.821); postoperative bleeding (OR, 2.80; 95% CI, 1.32-5.97); graft dysfunction (OR, 2.07; 95% CI, 1.21-3.53); primary graft failure (OR, 0.07; 95% CI, 0.01-0.33). CONCLUSION: Infection is a serious complication that continues to be difficult to control. Certain risk factors can be improved with careful management (kidney failure, pulmonary edema) or appropriate donor-recipient matching (initial dysfunction). Others, however, are inherent to the procedure (post-reperfusion syndrome, sex) or to immunosuppression, which acts as a true mediator of infection with regard to both its appearance and its clinical manifestation. PMID- 15901026 TI - [Stress, hemodynamic and immunological responses to inhaled and intravenous anesthetic techniques for video-assisted laparoscopic cholecystectomy]. AB - OBJECTIVE: Given the immunomodulatory effects of anesthesia and surgery, 2 anesthetic regimens in clinical use were compared to evaluate hemodynamic, stress, and immunologic response in patients undergoing laparoscopic cholecystectomy. PATIENTS AND METHODS: Randomized controlled trial in patients classified ASA I and scheduled for laparoscopic cholecystectomy. Patients were randomly assigned to the inhaled anesthetic group (13 anesthetized with propofol fentanyl-isoflurane) or the total intravenous anesthesia (TIVA) group (14 patients anesthetized with propofol-remifentanil). Patients in both groups received the muscle relaxant vecuronium. We assessed hemodynamic variables, cortisol levels, prolactin, interleukin 6, white cell and lymphocyte counts before, during (1 hour after induction) and after (24 hours and 7 days) surgery. RESULTS: Hemodynamic variables were stable in both groups. Significant changes in prolactin levels and markers of immune and inflammatory responses between baseline and later measurements occurred in both groups. Patients who received TIVA had no change in cortisol levels at any time during the study. The TIVA group had lower levels of cortisol than did the inhaled anesthesia group (TIVA, 207 [SD, 100] ng/mL; inhaled 293 [97] ng/mL; P<0.05)), higher neutrophil counts (TIVA, 75 [12.5]%; inhaled: 62 [20]%; P<0.05) and higher CD4+ T lymphocyte counts (TIVA, 53 [11.6]%; inhaled: 42 [17.6]%; P<0.001). CONCLUSION: Although both techniques afford hemodynamic stability, lower cortisol levels were observed with the application of TIVA with propofol-remifentanil. That would be the technique of choice for patients with compromised immune response. PMID- 15901027 TI - [Epidural obstetric analgesia, maternal fever and neonatal wellness parameters]. AB - OBJECTIVE: To study the relation between epidural analgesia and the development of maternal fever during labor and childbirth, and to determine the possible relation between that association and neonatal welfare and in the performance of tests to rule out sepsis in newborns. PATIENTS AND METHODS: Prospective study of all women who gave birth at Fundacion Hospital Alcorcon over a period of 3 years. All the women were offered epidural analgesia based on infusion of 0.0625% bupivacaine and 2 microg x mL(-1). Data collected were age, nulliparity, epidural analgesia infusion, induction of labor, uterine stimulation with oxytocin, type of birth, fetal weight, duration of dilation and expulsion, Apgar score (at 1 and 5 minutes), umbilical artery pH, and maternal temperature. RESULTS: Data for 4364 women were analyzed. Fever developed during labor in 5.7%; 93.7% of the fevers occurred in women receiving epidural analgesia (P<0.05). Logistic regression analysis revealed that independent risk factors for the development of fever were epidural analgesia (odds ratio [OR], 1.78; 95% confidence interval [CI], 1.05 3.04), nulliparity (OR, 2,929; 95% CI, 2.005-4.279), fetal weight (OR, 1.484; 95% CI, 1.102-2.001), and duration of labor (OR, 1.003; 95% CI, 1.003-1.004). No significant differences in Apgar score at 5 minutes or umbilical artery pH were found between the women with and without fever. Tests to rule out sepsis were ordered for 85.1% of the infants of mothers with fever after epidural analgesia. CONCLUSIONS: Epidural analgesia was associated with greater risk of developing fever in mothers giving birth, but that association had no repercussion on the neonatal wellness parameters studied. PMID- 15901028 TI - [Magnesium in anesthesia and postoperative recovery care]. AB - Magnesium is involved in many physiological processes and in the pathophysiology of many diseases that affect surgical patients. The incidence of hypomagnesemia in the perioperative setting is high and is sometimes underestimated, with important prognostic implications. Magnesium also has a variety of therapeutic indications in postoperative recovery care, obstetrics, cardiology, heart surgery, pain treatment, anesthesia, pneumology, etc. Magnesium's role in the organism and its pharmacological properties continue to be studied and new situations in which the ion plays a relevant part are being suggested. It has become essential for the anesthesiologist to understand the pharmacological, clinical, and physiological properties of magnesium. The present review aims to give a simple but complete overview of the physiological importance of the magnesium ion, the perioperative changes that occur, and its therapeutic applications in numerous clinical contexts. PMID- 15901029 TI - [Intracranial hypotension syndrome treated with an epidural blood patch]. AB - A 41-year-old woman was admitted to the internal medicine department to assess incapacitating postural headache. Clinical findings suggested the need for computed tomography and nuclear magnetic resonance scanning of the head, which led to a diagnosis of spontaneous intracranial hypotension syndrome. Later, isotopic cysternography and nuclear magnetic resonance imaging of the spine were used unsuccessfully to try to locate the cerebrospinal fluid leak that caused the syndrome. When conservative treatment proved ineffective, the pain clinic was called in to perform an epidural blood patch procedure. The patch led to an improvement in symptoms and the syndrome resolved completely after a second lumbar blood patch was used. PMID- 15901030 TI - [Subdural empyema due to Mycoplasma hominis after a cesarean section under spinal anesthesia]. AB - The isolation of Mycoplasma hominis in cultured biopsy material from a subdural empyema is a very rare finding. Likewise, subdural empyema complicating epidural anesthesia is an uncommon event after cesarean delivery. We report the case of a 32-year-old patient who presented a throbbing headache when standing 48 hours after undergoing cesarean delivery under spinal anesthesia. On the fifth day after surgery, the headache worsened, fever developed, and an abscess was detected at the abdominal wall; antibiotic treatment was prescribed. When fever and headache persisted and abdominal infection had been ruled out, nuclear magnetic resonance imaging of the head revealed subdural empyema. Emergency surgery to drain pus was carried out twice. Mycoplasma hominis was isolated from a blood-agar culture of the exudate. The patient recovered fully after combined surgical and antibiotic treatment. PMID- 15901031 TI - [Problems anchoring epidural catheters with Drain-Fix devices]. PMID- 15901032 TI - [Cardiac arrest during tracheal extubation in a woman undergoing surgery for pheochromocytoma]. PMID- 15901033 TI - [Tonsillectomy and refractory bronchospasm in a girl with Down's syndrome]. PMID- 15901034 TI - [Anesthesia in Wilson's disease]. PMID- 15901035 TI - [Hemostasis-altering drugs and techniques for regional anesthesia and analgesia: safety recommendations]. PMID- 15901036 TI - [On 2 cases of convulsions after use of ropivacaine for brachial plexus block]. PMID- 15901037 TI - [Diffuse idiopathic skeletal hyperostosis, Forestier and Rotes-Querol syndrome, and difficult spinal puncture]. PMID- 15901038 TI - [Dental care of patients with leukemia]. AB - Leukemias include a variety of acute and chronic malignant hematological diseases that require antineoplastic chemotherapy. Hematological stem cell transplantation allows an aggressive chemotherapy during which patients suffer from severe immunosuppression. Oral infections may cause serious complications during this immunosuppression. Therefore, professional diagnosis and elimination of oral infection foci must be carried out as early as possible before such treatment. Aggressive chemotherapy causes hyposalivation and as a consequence an increased risk for oral hard and soft tissue diseases. Adequate oral care before, during and after chemotherapy combined with a stem cell transplantation is necessary to prevent oral diseases and systemic complications of oral origin. PMID- 15901039 TI - [The penetration of various adhesives into early enamel lesions in vitro]. AB - The aim of the present study was to evaluate the penetration depth (PD) and the thickness of the oxygen inhibition layer (OIL) of a fissure sealant (Helioseal, Vivadent) and various adhesives (Heliobond, Excite, Vivadent; Resulcin, Merz; Solobond M, Voco; Prompt L-Pop, 3M-Espe) applied to enamel lesions in vitro. From 27 bovine teeth 54 enamel specimens were prepared and covered with nail varnish (control) thus obtaining three windows for treatment. After demineralisation (pH 5.0, 14 d) two of the windows were etched with phosphoric acid (20%, 5 s), whereas the third area served as control. The specimens were divided randomly into six groups (n = 9) and the respective adhesive was applied (90 s), either once or twice. Light-curing followed each application. Enamel slabs (perpendicular to the surface) were cut and studied after infiltration with a fluorescent low-viscous resin using confocal microscopy (CLSM). The image of the lesion was divided into two areas with different grey values. Lesion depths were calculated (ImageJ) from the surface to that point in the lesion where the grey value clearly changed to a darker grey value. The zone with the darker grey values marked the front of demineralisation. Mean lesion depths (+/- SD) after demineralisation were measured at 105 (+/- 21) microm. After single application, Resulcin [89 (+/- 22)%] and Helioseal [98 (+/- 6)%] had almost completely penetrated the lesion. Heliobond [126 (+/- 33)%] and Excite [184 (+/- 40)%] penetrated even deeper than the defined lesion. For Excite double application decreased the OIL significantly (p = 0.03; adjusted paired t-test). Adhesives are capable to penetrate artificial initial enamel lesions completely. Follow-up studies are needed to confirm this effect for natural lesions. PMID- 15901040 TI - [Care of a patient with an implant-borne, bar-retained prosthesis in the maxilla. Case report]. AB - A case presentation documents the prosthetic treatment of a denture in the upper jaw, and a conventional tooth--supported as well as a combined tooth--implant- supported fixed partial denture in the lower jaw. PMID- 15901041 TI - Comparative bioavailability/ bioequivalence of two different sertraline formulations: a randomised, 2-period x 2-sequence, crossover clinical trial in healthy volunteers. AB - An open-label, randomised, crossover single dose study, using 2 periods x 2 sequences, with a minimum washout period of 4 weeks, was conducted in order to assess the comparative bioavailability of two formulations of sertraline hydrochloride (CAS 79617-96-2) 100 mg tablets. Plasma samples were obtained at intake (baseline) and at +1 h, +2 h, +3 h, +4 h, +5 h, +6 h, +7 h, +8 h, +9 h, +12 h, +24 h, +48 h, +72 h and +96 h post administration. Sertraline plasma concentrations were determined by high pressure liquid chromatography with tandem mass detection (HPLC-MS/MS) and the lower limit of quantification was set at 100.15 pg/mL. Pharmacokinetic parameters used for bioequivalence assessment (AUClast, AUCinf and Cmax) were determined by non-compartmental analysis. Classical 90 % confidence intervals (90CI) were calculated for the overall sample, and for males and females separately, and gender effects were investigated using an appropriate model. The results showed that overall classical 90CI were 84.55-100.32 % for Cmax 86.96-98.68 % for AUClast, and 86.79 98.93 for AUCinf, that is, they were all within the predefined ranges for bioequivalence acceptance. Separate gender analysis showed very similar results for males and females when analysed independently, and no gender effects were detected in bioequivalence analysis (p > 0.05). It may be therefore concluded that the evaluated formulations are bioequivalent in terms of rate and extent of absorption. PMID- 15901042 TI - Single and multiple dose pharmacokinetic evaluation of a transdermal delivery system of imipramine hydrochloride. AB - The transdermal route provides an attractive alternative to the presently used peroral therapy with tricyclic antidepressants due to the avoidance of first-pass metabolism and the associated side effects. In this investigation an earlier developed transdermal delivery system (TDS) of imipramine hydrochloride (CAS 113 52-0; IMH) was evaluated with respect to dose proportionality at three different dose levels. Linearity was observed with the lower doses. For the prediction of in vivo plasma levels, various pharmacokinetic parameters such as alpha, beta, volume of distribution, and AUC0-infinity. were determined by single dose intravenous administration (2 mg/kg). The lowest dose was selected for the multiple dose study taking into consideration the issues of stability, safety, therapeutic range and linearity of pharmacokinetics. At all dose levels the experimental plasma values were significantly lower than predicted levels (p < 0.05) but 30-50 fold higher than the therapeutic range with no significant difference at different dose levels. The plasma levels obtained by repeated application were comparable to that obtained in the single dose study. In addition, IMH exhibited dose proportional pharmacokinetics at the higher doses (above 50 mg/day). The developed TDS was able to maintain steady-state plasma levels for the entire duration of the multiple dose study. PMID- 15901043 TI - Effects of adrenoceptor blocking drugs on cardiovascular responsiveness to passive orthostasis: a placebo-controlled double-blind study. AB - OBJECTIVE: To compare the acute effects of the beta-blocker propranolol (CAS 525 66-6), beta + alpha1-blocker carvedilol (CAS 72956-09-3) and alpha1-blocker tamsulosin (CAS 106463-17-6) on the cardiovascular responses to passive orthostasis. METHODS: The responses to passive orthostasis (tilt provocation at 60 degress for 8 min) were measured in normotensive healthy volunteers with finger-blood-pressure and whole-body impedance cardiography prior to drugs and three days after beginning the medications. The treatments were moderate oral doses of the beta-blocker propranolol HCl (40 mg twice daily, n = 7), (beta + alpha1-blocker carvedilol (12.5 mg once daily for two days and thereafter 25 mg daily, n = 7), alpha1-blocker tamsulosin HCl (0.4 mg once daily, n = 6), or placebo. The drugs were distributed in a randomised, double-blind fashion. RESULTS: When measured prior to the head-up tilt test, propranolol and carvedilol had decreased supine systolic arterial pressure (SAP) more than placebo, and heart rate (HR) more than placebo and tamsulosin. Propranolol had decreased also pulse wave velocity more than placebo. The cardiac index (CI), stroke index (SI) and systemic vascular resistance index (SVRI) were not changed with any drugs. During the head-up tilt, tamsulosin decreased SAP and SVRI and augmented CI but not SI. The SVRI response curve with tamsulosin differed from that with propranolol or carvedilol. Also, tamsulosin increased HR compared to the beta blocking drugs. The propranolol and carvedilol groups did not differ from each other in terms of any parameter. CONCLUSIONS: Tamsulosin significantly decreases SAP in the passive head-up tilt, indicating that it is not purely uroselective alpha1-blocker. The responses with tamsulosin clearly differ from those with propranolol and carvedilol, the responses with the latter two being practically equal. PMID- 15901044 TI - Bioequivalence study of two different tablet formulations of carvedilol in healthy volunteers. AB - OBJECTIVE: The aim of this study was to compare the extent and rate of absorption of two different carvedilol (CAS 72956-09-3) tablet formulations: 25 mg tablets, as the test formulation and the reference innovator product (25 mg tablets). METHODS: This study was designed as a single-dose, open-label, randomised, with a two-period and two-sequence crossover design, with blind determination of drug plasma concentration and a minimum 7-day washout period. Twenty-four healthy volunteers of both sexes were randomly assigned to treatment sequences. Carvedilol concentrations were determined in plasma samples obtained over a 24-h interval: baseline (pre-administration) and at 14 different times within the 24 h after administration. The analytical method, which used HPLC coupled with a MS/MS detector, was duly validated and the analytical assay was performed in compliance with Good Laboratory Practice (GLP). The limit of quantification (LOQ) was 0.50 ng/mL. Pharmacokinetic parameters representing the extent and/or rate of absorption (AUCinf, AUClast, and Cmax) were obtained. As secondary objective the tolerability of both formulations was also evaluated. RESULTS: The geometric mean of the test/reference formulations individual percent ratio was 98.14 % for AUCinf, 98.44 % for AUClast and 98.39 % for Cmax. The 90 % CI for the geometric mean of the individual ratio test/references formulations was 95.13 to 101.24 % for AUCinf, 95.23 to 101.76 % for AUClast, and 88.26 to 109.67 % for Cmax. CONCLUSIONS: The 90 % CI values obtained for AUCinf, AUClast, and Cmax are within the interval proposed by the EMEA/CPMP and the FDA as bioequivalence acceptance criteria, and consequently it can be conclude that the test formulation is bioequivalent with the reference formulation both in terms of rate and extent of absorption after single dose administration. The results from a previous pilot study allowed an optimal design for this trial. PMID- 15901045 TI - Bioequivalence study of two different coated tablet formulations of finasteride in healthy volunteers. AB - This study was conducted in order to assess the bioequivalence of two different coated tablet formulations containing 5 mg finasteride (CAS 98319-26-7). Twenty six healthy volunteers were enrolled in an open, randomised, crossover single dose study with 2 periods x 2 sequences and a minimum washout period of 7 days. Plasma samples were obtained over 24 h (at baseline, +0.5 h, +1 h, +1.5 h, +2 h, +2.5 h, +3 h, 3.5 h, +4 h, +4.5 h, +5 h, +6 h, +8 h, +10 h, +12 h, +16 h and +24 h after administration). Finasteride levels were determined by high-pressure liquid chromatography with tandem mass detection, HPLC-MS/ MS, (LOQ 0.50 ng/mL). Pharmacokinetic parameters used for bioequivalence assessment (AUClast and Cmax were main evaluation criteria, however, AUCinf was also analysed) were determined from the finasteride concentration data using non-compartmental analysis. The 90 % confidence intervals (obtained by ANOVA) were 86.31-98.69 for Cmax, 95.40 104.88 for AUClast and 96.20-105.81 for AUCinf that is, they were all within the predefined ranges. It may be therefore concluded that the evaluated formulations are bioequivalent in terms of rate and extent of absorption. PMID- 15901046 TI - In vitro anti-HIV potency of stampidine alone and in combination with standard anti-HIV drugs. AB - The purpose of the present study was compare the in vitro anti-HIV potency stampidine (CAS 217178-62-6), a novel aryl phosphate derivative of stavudine (CAS 3056-17-5), and drug combinations containing stampidine to the anti-HIV tency of the standard drugs zidovudine (CAS 30516-87-1), stavudine, lamivudine (CAS 134678 17-4), nelfinavir (CAS 159989-65-8), and nevirapine (CAS 129618-40-2) as well as their combinations. Stampidine inhibited the laboratory HIV-1 strain HTLV(IIIB) (B-envelope subtype) as well as the primary clinical HIV-1 isolates BR/92/025 (C envelope subtype) and BR/93/20 (F-envelope sub-type) with subnanomolar IC50 values. Stampidine was as effective as zidovudine against HTLV(IIIB) and BR/92/025 and 3-logs more effective than zidovudine against BR/93/20. Stampidine was more effective than stavudine, lamivudine, nelfinavir, and nevirapine against all three HIV-1 isolates. The combination of stampidine with zidovudine + lamivudine was more effective than the combination of nelfinavir or nevirapine with zidovudine lamivudine against all three HIV-1 isolates. The combination of stampidine with nelfinavir was more effective than zidovudine + lamivudine as well as the combination of zidovudine + lamivudine with nelfinavir. The combination of stampidine with lamivudine + nelfinavir was more effective than the combination of zidovudine with lamivudine + nelfinavir. The combination of stampidine with lamivudine + nevirapine was more effective than the combination of stavudine with lamivudine + nevirapine. These findings demonstrate that (a) stampidine, as well as its combinations with the standard anti-HIV drugs zidovudine, lamivudine, nelfinavir or nevirapine, are potent inhibitors of HIV-1 replication in human peripheral blood mononuclear cells, and (b) replacement of either zidcovudine, zidovudine+lamivudine or stavudine in 3-drug cocktails with stampidine resulted in greater anti-HIV potency in vitro. PMID- 15901047 TI - In vitro activities of thiadiazine derivatives against Leishmania amazonensis. AB - Ten thiadiazine derivatives were tested in vitro for antiparasitic effects against both extracellular promastigotes and intracellular amastigotes of Leishmania amazonensis. The results showed that the evaluated compounds exhibited a strong antiproliferative activity on all developmental stages of the parasite. The minimal inhibitory concentration and the 50 % effective concentration values against the promastigote were 2.1-5.1 microg/ml and 0.6-1.8 microg/ml, respectively. The tested compounds caused an irreversible inhibition of the promastigote growth either after 1 h of treatment with 10 microg/ml or after 24 h with 1 microg/ml. Also, the thiadiazine derivatives were active against amastigotes producing between 12 and 89 % of reduction of infection at 100 ng/ml. However, the compounds exhibited high toxicity and provoked inhibition of the phagocytosis in the murine host cell. PMID- 15901048 TI - Validation of a simple bioanalytical method for the determination of DRF-6196, a novel oxazolidinone, in mouse plasma: application to a single-dose pharmacokinetic study. AB - An isocratic simple, specific, sensitive and reproducible high performance liquid chromatography (HPLC) method was developed and validated for the estimation of DRF-6196, a novel oxazolidinone in mouse plasma. This method involves a simple liquid/liquid extraction of DRF-6196 and the internal standard (IS; chlorzoxazone, CAS 95-25-0) from plasma into dichloromethane/ethyl acetate mixture that was evaporated under nitrogen. The HPLC analysis was carried out on an Inertsil ODS 2 column using 0.01 mol/L potassium dihydorgen ortho phosphate (pH 3.2) and acetonitrile (65:35, v/v) as mobile phase. The eluate was monitored using an UV detector set at 266 nm. Ratio of peak area of analyte to IS was used for quantification of plasma samples. The retention time of DRF-6196 and IS were 8.2 and 11.1 min, respectively. The assay was linear (r2 > 0.999) in the concentration range 0.1-50 microg/ml. Absolute recovery for analyte and IS was > 94 % from mouse plasma. The lower limit of quantification (LLOQ) of DRF-6196 was 0.1 microg/ml. The inter- and intra-day precision in the measurement of quality control (QC) samples, 0.1, 0.3, 15.0 and 40.0 microg/ml, were in the range 3.64 to 9.51 % relative standard deviation (RSD) and 0.92 to 6.23 % RSD, respectively. Accuracy in the measurement of QC samples was in the range 88.15 to 106.05 % of the nominal values. The analyte and IS were stable in the stability studies viz., benchtop, autosampler and freeze/thaw cycles. The stability of DRF-6196 was established for 1 month at -80 degrees C. The assay method was successfully applied to a pharmacokinetic study of DRF-6196 in mice. PMID- 15901049 TI - Inflammatory markers in bronchoalveolar lavage fluid from human volunteers 2 hours after hydrogen fluoride exposure. AB - Fluoride has been in focus as a possible causal agent for respiratory symptoms amongst aluminium potroom workers for several decades. Previously, using bronchoalveolar lavage (BAL), we demonstrated airway inflammation in healthy volunteers 24 hours after exposure to hydrogen fluoride (HF). The objective of the present study was to examine early lung responses to HF exposure. Bronchoscopy with BAL was performed 2 hours after the end of 1-hour exposure to HE Significant reductions in the total cell number and the number of neutrophils and lymphocytes were observed in bronchoalveolar portion (BAP), whereas there were no significant changes in the bronchial portion (BP). Significantly decreased concentrations of beta2-MG, IL-6 and total protein were found in both BAP and BP. Additionally, IL-8 was significantly reduced in BP, and ICAM-1 and albumin were present in lower concentrations in BAP. Lung function measurements were not affected by HF exposure. These reported effects are presumably transitory, as many were not present in the airways 24 hours after a similar HF exposure. PMID- 15901050 TI - Safety pharmacology of sibutramine mesylate, an anti-obesity drug. AB - Sibutramine mesylate is a new anti-obesity drug. It is a crystalline salt of sibutramine developed to improve the solubility of sibutramine hydrochloride. Methanesulfonic acid was used as a salt-forming acid instead of hydrochloric acid, resulting in a greatly improved solubility of 1000 mg/mL in water. Sibutramine mesylate was administered orally to ICR mice, Sprague-Dawley rats, and beagle dogs at dose levels of 1.15, 3.45, and 11.50 mg/kg to measure its effects on the central nervous system (CNS), general behaviour, cardiovascular respiratory system and the other organ systems. Following administration of sibutramine mesylate, spontaneous locomotor activity was significantly increased from 120 min to 24 hours at 3.45 mg/kg and from 30 min to 24 hours at 11.50 mg/kg. Furthermore, there were a decrease in hexobarbital-induced sleep time, an increase in respiratory rate at 120 min, increases in intestinal transport capacity and gastric pH at 11.50 mg/kg, and decreases in gastric volume and total acidity at 3.45 and 11.50 mg/kg. However sibutramine mesylate caused no effects on general behaviour, motor coordination, body temperature, analgesia, convulsion, blood pressure, heart rate, electrocardiogram, cardiac functions of the isolated rat heart, isolated smooth muscles and renal function. Based on the above results, it was concluded that sibutramine mesylate caused effects on the spontaneous locomotor activity, hexobarbital-induced sleep time, respiration, gastrointestinal transport, and gastric secretion at a dose level of 3.45 mg/kg or greater but caused no effects on other general pharmacological reactions. PMID- 15901051 TI - Antigenotoxic effects of ascorbic acid against megestrol acetate-induced genotoxicity in mice. AB - The genotoxicity of megestrol acetate was studied in mouse bone marrow cells using sister chromatid exchanges (SCEs) and chromosomal aberrations (CAs) as parameters. Megestrol acetate (8.12, 16.25 and 32.50 mg/ kg of body weight) was injected intraperitoneally separately in different groups of animals. Both CAs and SCEs were statistically increased at 16.25 and 32.50 mg/kg of body weight. Our earlier in vitro studies show the generation of free oxygen radicals, by synthetic progestins responsible for the genotoxic damage. As the genotoxic effects of steroids can be reduced by natural products having antioxidant properties, and ascorbic acid possesses antioxidant activity, ascorbic acid (20, 40 or 60 mg/kg of body weight) administered together with megestrol acetate (32.50 mg/kg of body weight) significantly decreased CAs and SCEs, suggesting an antigenotoxic role of ascorbic acid against megestrol acetate-induced genotoxic damage in mice bone marrow cells. The antigenotoxic effect was clearly dose dependent. The highest protective effect was observed at 60 mg/kg body weight of ascorbic acid treated with 32.50 mg/kg body weight of megestrol acetate. PMID- 15901052 TI - Protective effects of metformin treatment on the liver injury of streptozotocin diabetic rats. AB - Metformin is a biguanide derivate used as an oral hypoglycaemic drug in diabetics. The aim of this study was to examine the histological and biochemical effects of metformin in streptozotocin (STZ)-treated rats. The animals were rendered diabetic by intraperitoneal injection of 65 mg/kg STZ. Fourteen days later, metformin was given at 25 mg/kg by gavage, daily for 28 days, to STZ diabetic rats and a control group. In the STZ-diabetic group, some degenerative changes were observed by light microscopic examination. But the degenerative changes were decreased in the STZ-diabetic group given metformin. In the STZ diabetic group, blood glucose levels, serum alanine and aspartate transaminase (ALT and AST) activities, total lipid levels, and sodium and potassium levels increased, while body weight, serum magnesium levels and liver glutathione (GSH) levels decreased. In the STZ-diabetic group given metformin, blood glucose levels, serum ALT and AST activities, total lipid, and sodium and potassium levels decreased, and liver GSH and serum magnesium levels increased. As a result of all the morphological and biochemical findings obtained, it was concluded that metformin has a protective effect against the hepatotoxicity produced by STZ diabetes. PMID- 15901053 TI - Neuroprotective effects of Withania somnifera on 6-hydroxydopamine induced Parkinsonism in rats. AB - 6-Hydroxydopamine (6-OHDA) is one of the most widely used rat models for Parkinson's disease. There is ample evidence in the literature that 6-OHDA elicits its toxic manifestations through oxidant stress. In the present study, we evaluated the anti-parkinsonian effects of Withania somnifera extract, which has been reported to have potent anti-oxidant, anti-peroxidative and free radical quenching properties in various diseased conditions. Rats were pretreated with 100, 200 and 300 mg/kg b.w. of the W. somnifera extract orally for 3 weeks. On day 21, 2 microL of 6-OHDA (10 microg in 0.1% in ascorbic acid-saline) was infused into the right striatum while sham operated group received 2 microL of the vehicle. Three weeks after 6-OHDA injections, rats were tested for neurobehavioral activity and were killed 5 weeks after lesioning for the estimation of lipidperoxidation, reduced glutathione content, activities of glutathione-S-transferase, glutathione reductase, glutathione peroxidase, superoxide dismutase and catalase, catecholamine content, dopaminergic D2 receptor binding and tyrosine hydroxylase expression. W. somnifera extract was found to reverse all the parameters significantly in a dose-dependent manner. Thus, the study demonstrates that the extract of W. somnifera may be helpful in protecting the neuronal injury in Parkinson's disease. PMID- 15901054 TI - Soy isoflavones inhibits the genotoxicity of benzo(a)pyrene in Swiss albino mice. AB - Dietary factors are considered important environmental risk determinants for various diseases. Isoflavones are one of the biologically active polyphenolic plant constituents that possess potential chemopreventive properties against a wide variety of chronic diseases. In the present study we have evaluated the antimutagenic potential of soy isoflavones against benzo(a)pyrene (B[a]P) (125 mg/ kg) induced genotoxicity in Swiss albino mice. The effect of soy isoflavones was studied by in vivo bone marrow chromosomal aberration and micronuclei induction test. Using an alkaline unwinding assay we monitored the DNA strand breaks. Two doses of soy isoflavones (20 and 40 mg/kg b.wt) were given orally for seven days prior to the administration of B[a]P. Soy isoflavone inhibited the genotoxicity of B[a]P in terms of chromosomal aberration and micronucleus formation. DNA strand break levels in only B[a]P treated group remained significantly high from the control values (P < 0.001). The pretreatment of soy isoflavone showed gradual reduction in strand breaks significantly (P < 0.001) and dose dependently. Soy isoflavone pretreatment also decreased cytochrome P450 (CYP) content. The activity of CYP was also decreased dose dependently by pretreatment with soy isoflavone. The chemopreventive effect of soy isoflavone on the inhibition of CYP activity and DNA integrity mediate the possible mechanism of inhibition of genotoxicity. PMID- 15901055 TI - Designation of substances as skin sensitizing chemicals: a reply. PMID- 15901056 TI - ECHN honors cancer survivors with fun, food and inspirational stories. AB - A nostalgia theme was fully explored by Eastern Connecticut Health Network (ECHN), Manchester, Conn., in its celebration of Cancer Survivors Day, June 6. The observance is sponsored by the national Cancer Survivors Day organization. This year more than 700 facilities across the country observed the occasion. PMID- 15901057 TI - Publicist's advice: hire a PR firm before you need one. AB - Tom Rees, self-confessed former PR representative for the University of Colorado Health Sciences Center, addresses the value healthcare organizations can find in public relations consultation. PMID- 15901058 TI - Parma and Prince William hospitals deliver magazines with purpose. AB - Two quarterly publications are featured here for your consideration. One is Informed, the newly re-designed magazine by Parma Community General Hospital, Parma, Ohio. Also, we present Healthy Perspective magazine, published quarterly Prince William Health System, Manassas, Va. PMID- 15901059 TI - OSF Saint Anthony finds web site important as a recruitment tool: consistent monitoring of site reveals reader interest. AB - OSF Saint Anthony Medical Center, Rockford, Ill., has found its web site, www.osfhealth.com, to be a highly valuable tool for recruiting employees. The 254 bed acute care center maintains its own web site in cooperation with its parent organization, OSF Healthcare System, Peoria, Ill. PMID- 15901060 TI - Hospitals target-marketing wound centers to build profit statements. AB - Wound care centers add to the hospital's' profits as well as serving as a marketing vehicle for other services. Learn how the program is promoted by Shawnee Mission Medical Center, Shawnee Mission, Kan., Stamford Hospital, Stamford, Conn., and Good Samaritan Hospital, Suffern, N.Y. PMID- 15901061 TI - Meridian health annual report focuses on what is 'brand new'. AB - The 2003-2004 annual report by Meridian Health, Neptune, N.J., is titled brand new. The high-end publication was produced especially for the healthcare system's employees, board members, shareholders and volunteers. PMID- 15901062 TI - Detroit's Henry Ford Health System awarded Foster G. McGaw Prize. AB - Henry Ford Health System, Detroit, is named winner of the prestigious Foster G. McGaw Prize for excellence in community service. The organization received 100,000 dollars from the sponsors, the American Hospital Association, The Baxter International Foundation and the Cardinal Health Foundation. PMID- 15901064 TI - Differentiation of the human mesenchymal stem cells derived from bone marrow and enhancement of cell attachment by fibronectin. AB - The ability of human mesenchymal stem cells (hMSC) to differentiate into osteoblasts was examined through the use of osteogenic induction medium (MSCOIM) cultures. hMSC first attached to the dish surface and exhibited fibroblast-like spindle shapes, and after proliferation, formed cuboidal shapes. Calcium assays and the use of von Kossa and alizarin red S staining showed that hMSC were capable of mineralization when cultured in MSCOIM. Gene expressions of Cbfa-1 and BMP-4, which are markers for osteogenic differentiation, were also increased during the hMSC differentiation into osteoblasts. When compared to albumin (Alb) coated dishes, microscopic observation documented enhanced cell attachment and spreading when hMSC were cultured on fibronectin (FN)-coated dishes. Adherent cell numbers also exhibited a greater increase on the FN-coated dishes during earlier culture stages than that seen for the Alb-coated dishes. These findings suggest that hMSC have the capability to differentiate into osteoblasts and that FN can stimulate the attachment and spreading of the hMSC. PMID- 15901063 TI - Relationship between Porphyromonas gingivalis, Epstein-Barr virus infection and reactivation in periodontitis. AB - The aim of this study was to assess the relationship-between Porphyromonas gingivalis, Epstein-Barr virus (EBV) infection and reactivation in periodontitis using real-time PCR. The mean proportion of P. gingivalis cells to total bacterial cells in the saliva from EBV-positive periodontitis patients was significantly higher than that in saliva from EBV-negative patients. An EBV positive B-cell line was used to determine whether P. gingivalis sonicate induced reactivation of EBV, using real-time PCR to measure the virus genome in the culture medium. A significant increase in EBV numbers was observed after the stimulation with P. gingivalis sonicate. These findings suggest that the interaction between EBV and P. gingivalis is bi-directional, with EBV reactivation suppressing host defenses and permitting overgrowth of P. gingivalis, and P. gingivalis having the potential to induce EBV reactivation. PMID- 15901065 TI - The effect of gamma radiation on enamel hardness and its resistance to demineralization in vitro. AB - Given the importance of sterilizing human teeth before using them in research, the effects of a 25 kGy dose of gamma radiation on the mechanical properties of enamel and its resistance to demineralization were evaluated. Thirty human teeth were sectioned longitudinally, and while one half of each tooth was irradiated, the other half was kept as a control. Abraded and unabraded enamel slabs were obtained from these halves. The surface microhardness (SMH) of abraded slabs of irradiated and non-irradiated enamel was determined to evaluate the effect of radiation on enamel structure. Further, both abraded and unabraded slabs, either irradiated or non-irradiated, were submitted to a pH-cycling model to evaluate enamel resistance to demineralization, which was quantified by mineral loss (deltaZ) using cross-sectional microhardness. The data for SMH and deltaZ were statistically analyzed by t-test and ANOVA, respectively. The difference in enamel SMH between slabs from irradiated teeth and non-irradiated teeth was not statistically significant (P > 0.05). The abraded enamel slabs showed higher values of deltaZ than unabraded enamel slabs (P < 0.05), but the irradiation effect was not statistically significant (P > 0.05). The results suggest that the medical gamma radiation dose of 25 kGy does not affect either enamel hardness or its resistance to demineralization. PMID- 15901066 TI - Esthetic perception and psychosocial impact of developmental enamel defects among Malaysian adolescents. AB - The aim of this study was to investigate the prevalence and psychosocial impact of enamel defects among 16-year-old school children on the island of Penang. The data were collected through a self-administered questionnaire survey and an oral examination, using the Modified Developmental Defects of Enamel Index (FDI, 1992). In all, 1024 subjects were selected using a multistage random sampling technique. About two-thirds of the sample (67.1%) had at least one tooth affected by enamel defects. Enamel opacities accounted for 85.6% of the total condition. Diffuse-type opacity predominated (63.5%). Among subjects who expressed dissatisfaction, 18.8% reported covering their mouths when smiling, 8.7% avoided going out with friends and 39.1% had consulted their dentists. About 17% of the subjects reported that their parents had complained about the color of their front teeth but only 5.7% had experienced being teased by their friends about the problem. Two-thirds of the subjects were affected by enamel defects involving at least one tooth; however, the esthetic perception and psychosocial impact of those affected were minor. PMID- 15901067 TI - Extracellular matrix molecules in chronic obstructive sialadenitis: an immunocytochemical and Western blot investigation. AB - The exact pathomechanism of inflammation progress and fibrosis in chronic sialadenitis is unknown. Connective tissue growth factor (CTGF), matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) have been implicated in the pathogenesis of various fibrotic conditions. These factors are thought to be essential in the regulation of extracellular matrix turnover and the development of tissue fibrosis. In the present study, the expression of CTGF, MMP-2, -3, -9, -13 and TIMP-3 was examined in chronic obstructive sialadenitis. Tissue samples of 13 patients with chronic sialadenitis of the submandibular gland associated with sialolithiasis and 4 normal tissue samples of the submandibular gland were analyzed immunohistochemically and by Western blot analysis. An intense CTGF immunoreactivity was observed in the ductal system of inflamed salivary glands, whereas in normal glands no reactivity or a very low CTGF immunoreactivity was present. Immunohistochemical studies revealed a low to strong reactivity of MMP-2, -3, -9, -13, and TIMP-3 in the ductal system, in acinar cells and in lymphomonocytic infiltrates in normal and inflamed tissues. The expression of MMP-2, -3, -9, -13, and TIMP-3 was confirmed by Western blotting in all cases. Over-expression of CTGF in chronic obstructive sialadenitis suggests that this factor may play a role in glandular fibrosis. However, the physiological role of MMP-2, -3, -9, -13, and TIMP-3 in normal glands, as well as their possible role in inflammation progress and fibrosis in chronic obstructive sialadenitis, remains to be elucidated. PMID- 15901068 TI - Effect of noble metal adhesive systems on bonding between an indirect composite material and a gold alloy. AB - In this study, the bond strength between an indirect composite and a gold alloy was determined for the purpose of evaluating noble metal bonding systems. A single liquid primer designed for conditioning noble metal alloys (Infis Opaque Primer) and tri-n-butylborane-initiated adhesive resins (Super-Bond C & B), with or without the powder component, were assessed. Cast gold alloy disks (Casting Gold type IV) were air-abraded with alumina, followed by six surface preparations, and were then bonded with a light-activated composite material (New Metacolor Infis). Shear testing was performed both before and after thermocycling for evaluation of bond durability. The results showed that three primed groups improved post-thermocycling bond strengths compared to each of the corresponding unprimed groups (P < 0.01). The bond strength was reduced for all six groups by the application of thermocycling (P < 0.01). After thermocycling, the group primed with the Infis Opaque Primer material and bonded with the Super-Bond C & B resin exhibited the greatest bond strength (23.4 MPa). The Infis Opaque Primer and Super-Bond bonding system increased the post-thermocycling bond strength of the control group by a factor of approximately ten. This simple technique is applicable in the fabrication of composite veneered restorations and cone telescope dentures. PMID- 15901069 TI - Correlation study on oral health and electrocardiogram abnormalities. AB - The purpose of this study was to investigate the association between periodontal conditions and electrocardiogram test results that were obtained to screen for coronary heart disease risk factors. The present study included a total of 578 subjects who underwent annual medical check-ups at the Total Health Care Center in Otsu, Shiga Prefecture, Japan. To calculate the odds ratios for the electromyography abnormalities, we performed a logistic regression analysis for the oral examination, electrocardiogram, and blood analysis data. The crude odds ratio was obtained by a logistic regression analysis of age, sex, number of missing teeth, number of filled teeth, simplified oral hygiene index, community periodontal index, and blood analysis factors and results indicated there was a statistically significant correlation with the prevalence of electrocardiogram abnormalities. However, electrocardiogram abnormalities have a strong correlation with demographic factors such as sex and age. Therefore the experimental factors representing oral status were reexamined after the odds ratios were adjusted for age and sex. As a result of this adjustment, the new odds ratios that were determined indicated that there were no correlations between the oral factors and the prevalence of electrocardiogram abnormalities. PMID- 15901070 TI - Congenital absence of the permanent canines: a clinico-statistical study. AB - We report here a clinico-statistical study of congenital absence of the permanent canines (CAPC). Sixty-five cases (22 men and 43 women) of CAPC were found in the files of 35,927 outpatients, an incidence of 0.18%. Thirty-seven cases had single absence of the canine and 28 cases had multiple absences. There were 42 cases in the maxilla, 17 in the mandible, and 6 in both the maxilla and the mandible. In the single absence cases, there was a tendency of occurrence at the left side in the maxilla, and the right side in the mandible. There were 39 CAPC cases without absence of other permanent teeth. Several complicated dental anomalies were seen such as persistence of deciduous teeth, congenital absence of other permanent teeth, microdontia, malposition, and complete or incomplete impaction (excluding the third molars). The complication rate of other permanent tooth absences was higher in the cases with CACP in the maxilla than in the mandible. PMID- 15901071 TI - Odontogenic cysts: a clinical study of 90 cases. AB - The maxillofacial region is affected by a greater number of cysts than any other part of the body. In this study, 90 odontogenic cysts were collected from 90 patients over a five-year period. Patients with radicular cysts, dentigerous cysts and odontogenic keratocysts were further analyzed with regard to age, sex and anatomical distribution. Using the histological classification of the World Health Organization, 53 cases (59%) were classified as radicular cysts, 24 (27%) as keratocysts and 13 (14%) as dentigerous cysts. Radicular cysts occurred most frequently in the anterior region of the maxilla, odontogenic keratocysts in the ramus and angular region of the mandible, and dentigerous cysts in the mandible. No recurrences were observed during the limited follow-up period. PMID- 15901072 TI - Cleido cranial dysplasia: report of a family. AB - A family case of Cleidocranial Dysplasia is presented. A mother and two adolescent girls were examined. In all three cases, a radiological series was performed over the entire body. Generalized dysplasia in bones, prolonged retention of primary teeth, and delayed eruption of permanent, as well as supernumerary teeth was diagnosed. The citogenetic study with GTG band showed normal 46, XX. Bilateral audiometry in the mother demonstrated a mild to moderate hypoacustic condition. Radiological findings are presented and the importance of early diagnosis is discussed. PMID- 15901073 TI - [Early progress in water]. PMID- 15901074 TI - [The only dependable factor is the unreliability.-- children in alcoholic families]. PMID- 15901075 TI - [Conducting conversation with difficult patients and their relatives]. PMID- 15901076 TI - [Without diet children with phenylketonuria are threatened by intellectual decline/health insurance bound not to pay anymore for special nutrition]. PMID- 15901077 TI - [Fetal sensation]. PMID- 15901078 TI - [Allergies in the 1st life year. The Allergy Prevention Guideline 2004 of the Allergy Prevention Action Alliance (abap)]. PMID- 15901079 TI - [How children with bronchial asthma cope with their disease]. PMID- 15901080 TI - [Hotel Hospital: curse and blessings of rooming-in]. PMID- 15901081 TI - Goals and goal setting. AB - Once you decide upon your personal definition of what a high quality of life is, set goals accordingly to achieve that objective. This is accomplished by tracking the key performance indicators of your practice to gain a statistical backing, and then setting, documenting and achieving your goals through the concept of Goal Curves. In doing so, you reap benefits in not only your practice but also your personal life. PMID- 15901082 TI - Choice of a dental implant system. AB - Many dentists are bewildered by the intricacies and complexities of dental implants. They are constantly besieged by product advertisements and can find it difficult to choose which systems to work with. Some dentists are so intimidated by the subject that they choose to avoid getting involved with implants and instead stick to traditional tooth replacement systems. By breaking implants down into 4 main components, the body, collar, connection, and restorative post, it is easier to understand the structure and function of dental implants. Each portion should be designed to achieve certain objectives. Once these structural components are understood, it is easier to compare and contrast differing implant systems. PMID- 15901083 TI - Esthetic restoration of tooth structure using a nanofill composite system. PMID- 15901084 TI - The use of a mineralized allograft for sinus augmentation: an interim histological case report from a prospective clinical study. AB - A prospective clinical research study of various graft materials used for the augmentation of human maxillary sinuses is currently in progress at New York University Department of Implant Dentistry. This interim case report describes the healing response after a sinus augmentation procedure using a new mineralized cancellous bone allograft. The results after 9 months of healing demonstrated a vital bone content of 25.2% in the grafted sinus, as ascertained from a trephine core taken from the superior aspect of the lateral window area. Although the vital bone requirement for implant survival in an augmented sinus is unknown, the 25.2% vital bone demonstrated in this case compares favorably with that reported in the literature for other augmentation materials, including xenografts, alloplasts, and autogenous bone. PMID- 15901085 TI - Extraction-site ridge preservation. PMID- 15901086 TI - 'X' marks the spot. PMID- 15901087 TI - Extreme makeovers. PMID- 15901088 TI - Informed consent: a stepping stone in risk management. AB - This article discusses 1 facet of risk management: informed consent. It provides a basis from which the concept of risk management has evolved to the current understanding of informed consent. It develops the concept of informed consent through discussion of the elements that make up this integral part of risk management. It should be noted that, as with any legal concept, local legal counsel should be consulted, because state law is it applies to informed consent may vary. PMID- 15901089 TI - A forum for introducing new products into the dental school curriculum. PMID- 15901090 TI - Review and analysis of inhalation dosimetry methods for application to children's risk assessment. AB - Young children have a greater ventilation rate per body weight or pulmonary surface area as compared to adults. The implications of this difference for inhalation dosimetry and children's risk assessment were evaluated in runs of the U.S. Environmental Protection Agency (U.S. EPA) 1994 reference concentration (RfC) methodology and the ICRP 1994 inhalation dosimetry model. Dosimetry estimates were made for 3-mo-old children and adults for particles and Category 1 and 2 reactive gases in the following respiratory-tract regions: extrathoracic (ET), tracheobronchial (BB), bronchioles (bb), and pulmonary (PU). Systemic dosimetry estimates were made for nonreactive (Category 3) gases. Results suggest similar ET dosimetry for children and adults for all types of inhaled materials. BB dosimetry was also similar across age groups except that the dosimetry of ultrafine particles in this region was twofold greater in 3-mo-old children than in adults. In contrast, the bb region generally showed higher dosimetry of particles and gases in adults than in children. Particle dose in the PU region was two- to fourfold higher in 3-mo-old children, with the greatest child/adult difference occurring for submicron size particles. Particulate dosimetry estimates with the default RfC methodology were below those found with the ICRP model for both adults and children for submicrometer sized particles. There were no cases in which reactive gas dosimetry was substantially greater in the respiratory regions of 3-mo-old children. Estimates of systemic dose of Category 3 gases were greater in 3-mo-old children than in adults, especially for liver dose of metabolite for rapidly metabolized gases. These analyses support the approach of assuming twofold greater inhalation dose in children than adults, although there are cases in which this differential can be greater and others where it can be less. PMID- 15901091 TI - Contaminant loads and hematological correlates in the harbor seal (Phoca vitulina) of San Francisco Bay, California. AB - An expanding body of research indicates that exposure to contaminants may impact marine mammal health, thus possibly contributing to population declines. The harbor seal population of the San Francisco Bay (SFB), California, has suffered habitat loss and degradation, including decades of environmental contamination. To explore the possibility of contaminant-induced health alterations in this population, blood levels of polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (DDE), and polybrominated diphenyl ethers (PBDEs) were quantified in free-ranging seals; relationships between contaminant exposure and several key hematological parameters were examined; and PCB levels in the present study were compared with levels determined in SFB seals a decade earlier. PCB residues in harbor seal blood decreased during the past decade, but remained at levels great enough that adverse reproductive and immunological effects might be expected. Main results included a positive association between leukocyte counts and PBDEs, PCBs, and DDE in seals, and an inverse relationship between red blood cell count and PBDEs. Although not necessarily pathologic, these responses may serve as sentinel indications of contaminant-induced alterations in harbor seals of SFB, which, in individuals with relatively high contaminant burdens, might include increased rates of infection and anemia. PMID- 15901092 TI - PCBs and TCDD, alone and in mixtures, modulate marine mammal but not B6C3F1 mouse leukocyte phagocytosis. AB - Increasing evidence has supported the general hypothesis that organochlorines (OC) can produce immunotoxic effects in marine mammals. One important innate defense mechanism is phagocytosis, the ability of cells to ingest extracellular macromolecules. The present study is aimed at characterizing the immunomodulatory potential of mixtures of OCs on phagocytosis compared to that of individual compounds in different species of marine mammals and mice, the traditional model to study mammalian immunotoxicity. The ability of peripheral blood neutrophils and monocytes to engulf fluorescent microspheres was evaluated using flow cytometry. The immunomodulatory effects of three non-coplanar polychlorinated biphenyl (PCB) congeners, 138, 153, 180, one coplanar PCB, 169, as well as 2,3,7,8-TCDD, and all possible mixtures (26) were tested upon in vitro exposure. All species were not equally sensitive to the adverse effects of OCs on either neutrophils or monocytes phagocytosis. With the exception of harbor seals, all mixtures that significantly modulated neutrophil or monocyte phagocytosis contained at least one non-coplanar PCB. Regression analysis revealed that the non-coplanar congeners, more than the coplanar congeners, explained the variability in phagocytosis. Dendrograms revealed that phylogeny could not predict immunotoxicity. The currently used toxic equivalency (TEQ) approach and the traditional mouse model both failed to predict experimentally induced immunomodulatory effects in marine mammals tested, leading us to question the reliability of both TEQs and mouse model in risk assessment of OC mixtures. Testing the relative sensitivity to immunomodulatory effects of contaminants and contaminant mixtures between different species of marine mammals may have important implications for risk assessment as well as conservation and management strategies. PMID- 15901093 TI - Iron release and oxidative DNA damage in splenic toxicity of aniline. AB - Mechanisms by which aniline produces selective toxicity to the spleen are not well understood. Previously, studies showed that aniline exposure induces lipid peroxidation and protein oxidation in the spleen. The present study was aimed to determine the release of free iron and oxidative DNA damage in the spleen following aniline exposure. To achieve this, male SD rats were orally administered 1 mmol/kg/d aniline for 7 d, while controls received the vehicle only. Total splenic iron content showed a significant increase of 200% in the aniline-treated rats, whereas free iron (low-molecular-weight chelatable iron) showed a marked increase of 375% in comparison to controls. Oxidative DNA damage, measured in terms of 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, showed a remarkable increase of 83% in the aniline-treated rats. These results suggest an association between release of free iron and oxidative DNA damage, which could lead to mutagenic and/or carcinogenic responses in the spleen. PMID- 15901094 TI - In vitro conjugation of ethanolamine with fatty acids by rat liver subcellular fractions. AB - Previous studies from our laboratory have shown the enzymic formation of fatty acid (FA) conjugates of xenobiotic alcohols and amines. In the present study, the formation of FA conjugates of a bifunctional compound, ethanolamine was investigated by incubating [1-14C]oleic acid (1 mM) with ethanolamine (25 mM) at 37 degrees C in the presence of various rat liver subcellular fractions. The resultant product (or products) was separated by thin-layer chromatography (TLC) and the radioactivity corresponding to the relative flow of fatty acid amide was determined. Under similar conditions, formation of ethanolamides of palmitic, stearic, linoleic, linolenic, and arachidonic acids were also examined. The formation of ethanolamine conjugate with oleic acid was found to be 16.3 nmol/h/mg protein as compared to 6.7, 6.2, 8.1, 8.3, and 7.6 nmol/h/mg protein for palmitic, stearic, linoleic, linolenic, and arachidonic acids, respectively. The formation of oleoyl ethanolamide was found to be 18.9, 40.1, 65.9, and 0.3 nmol/h/mg protein in postnuclear, mitochondrial, microsomal, and cytosolic fractions, respectively. Mass spectrometric and nuclear magnetic resonance spectroscopic data of the TLC-purified product confirm the formation of oleoyl ethanolamide, and amidation appeared to be a preferred reaction over esterification. The results of this study suggest that the enzyme responsible for the amidation of fatty acids resides mainly in the microsomal fraction of the liver, and that oleic acid is a better substrate than other fatty acids used in the present study. PMID- 15901095 TI - A comparison of the potency of newly developed oximes (K005, K027, K033, K048) and currently used oximes (pralidoxime, obidoxime, HI-6) to reactivate sarin inhibited rat brain acetylcholinesterase by in vitro methods. AB - The potency of newly developed and currently used oximes to reactivate sarin inhibited acetylcholinesterase was evaluated using in vitro methods. A rat brain homogenate was used as a source of acetylcholinesterase. Significant differences in reactivation potency among all tested oximes were observed. Although the ability of newly developed oximes to reactivate sarin-inhibited acetylcholinesterase does not reach the reactivating potency of the oxime HI-6, the oxime K033 seems to be a more efficacious reactivator of sarin-inhibited acetylcholinesterase than other currently available oximes (pralidoxime, obidoxime) at concentrations (10(-5)-10(-4)M) corresponding to recommended doses in vivo. The results of our study also confirm that the reactivation potency of the tested reactivators depends on many factors, such as (1) the number of pyridinium rings, (2) the number of oxime groups and their position, and (3) the length and the shape of the linkage bridge between pyridinium rings. PMID- 15901096 TI - Management of herbicide-tolerant oilseed rape in Europe: a case study on minimizing vertical gene flow. AB - The potential commercialization of genetically modified herbicide-tolerant (GMHT) oilseed rape in Europe raises various concerns about their potential environmental and agronomic impacts, especially those associated with the escape of transgenes. Pollen of oilseed rape can be dispersed in space, resulting in the fertilization of sympatric compatible wild relatives (e.g. Brassica rapa) and oilseed rape cultivars grown nearby (GM and/or non-GM Brassica napus). The spatial and temporal dispersal of seeds of oilseed rape may lead to feral oilseed rape populations outside the cropped areas and oilseed rape volunteers in subsequent crops in the rotation. The incorporation of a HT trait(s) may increase the fitness of the recipient plants, making them more abundant and persistent, and may result in weeds that are difficult to control by the herbicide(s) to which they are tolerant. Vertical gene flow from transgenic oilseed rape to non GM counterparts may also have an impact on farming and supply chain management, depending on labelling thresholds for the adventitious presence of GM material in non-GM products. Given the extent of pollen and seed dispersal in oilseed rape, it is obvious that the safe and sound integration of GMHT oilseed rape in Europe may require significant on-farm and off-farm management efforts. Crucial practical measures that can reduce vertical gene flow include (1) isolating seed production of Brassica napus, (2) the use of certified seed, (3) isolating fields of GM oilseed rape, (4) harvesting at the correct crop development stage with properly adjusted combine settings, (5) ensuring maximum germination of shed seeds after harvest, (6) controlling volunteers in subsequent crops, and (7) keeping on-farm records. The implementation of the recommended practices may, however, be difficult, entailing various challenges. PMID- 15901097 TI - Gene flow scenarios with transgenic maize in Mexico. AB - Maize diversity is widespread in Mexico and it has been stewarded by campesinos in small communities until the present. With the arrival of transgenic maize, the objective of this study is to analyze possible scenarios that could result if genetically modified maize were not regulated and openly available in Mexico. By applying a simple logistic model based on the conditions of maize production in Mexico, the dispersion of transgenic maize in different situations within fields of farmers is described. In traditional open systems of freely exchanged seed within communities it is concluded that the most likely outcome of GM maize release is the incorporation of transgenes in the genome of Mexican germplasm and possibly in that of teosinte. PMID- 15901099 TI - [New health care: the hospital to come]. PMID- 15901098 TI - Diversity of alternative hosts of maize stemborers in Trans-Nzoia district of Kenya. AB - Genetically-engineered (GE) crops such as those expressing insecticidal Bacillus thuringiensis (Bt) toxin genes have the potential to greatly reduce the use of broad spectrum insecticides and increase crop productivity. However, development of resistance by the target insect species is an important consideration in the deployment of this strategy. In areas where GE crops are deployed on a large scale, current resistance management strategies rely on a 'refuge strategy', comprising the incorporation of a certain proportion of non-GE plants in the agro ecosystems, to conserve susceptible individuals of the target pests. In the USA, simulation models indicate that at least 20% of the crop should be non-Bt plants. In Africa, the target lepidopteran stemborers attack a wide range of wild grass species as well as cultivated cereal crops. Wild grasses generally occur in the vicinity of maize and other cereal fields, and may provide a refuge if GE crops are in the farming systems. To assess the quality of these grasses as refuges, it is critical to obtain information about their size and spatial distribution. In this study, we have assessed the abundance and diversity of alternative refuge of stemborers, mainly wild grasses occurring in the proximity of maize fields, in Trans-Nzoia district, one of the most important maize growing areas in Kenya. The proportion of wild host plants relative to maize was found to decline from 100% during the non-cropping season to <8% during the maize-growing season. The Shannon-Weaver diversity index indicated high variation in the diversity of wild hosts of stemborers between agro-ecological zones in the district. The results of this study are discussed in light of the possible role that wild host plant species might play in stemborer resistance management following the introduction of Bt maize. PMID- 15901100 TI - [Use of oral anticholinergic therapy in children under 1 years of age with high risk bladder]. AB - THE AIM: To assess the effectiveness and safety of the treatment with oral anticholinergic agents (Oxybutin clorure) in patients under 1 year old, and who aree carriers of high risk bladder secondary to neurological illness as well as no neurological one. MATERIAL AND METHODS: Since 1989, we have indicated treatment with anticholinergic agents to 16 patients: 9 patients had neurogenic bladder secondary to: myelomeningocele (n=7) and sacrocoxigeal teratoma (n=2). Others 7 patients had non neurogenic bladder secondary to: posterior urethral valvulas (n=1), valvula-like syndrome (n=4), post-surgery of neonatal giant bladder diverticulum (n=1) and Prune-Belly syndrome (n=1). The urodinamic study was performed during the first six months of life, being "high risk bladder" defined according to the parameters of compliance vesical and pressure of leak at point (PER). Five of the patients showed neonatal cronic renal failure (CRF), who were treated by cutaneus temporary derivation. All patients at treatment with anticholinergic agents at a 0.2 mg/kg/day dose was established; other early adjunctive treatment prior to the closure of the urinary derivation in children with CRF(n=5); or as a part of the conservative treatment (n=3), alone or associate to intermittent bladder catheterization (IBC) (n=8). During the treatment with anticholinergic agents, the cardiac frequency was controlled by EKG registration in 6 patients, being the rest of the children clinicaly controled (skin colour, mouth dryness, cardiac frequency and intestinal function). RESULTS: In all the cases, the minimum duration of the treatment was one year, until the functional stabilization of the urinary tract. It is to underline the absence of secondary complications which would have caused the suspension or the reduction of the treatment at long term. Conventional studies of urologicals image and urodinamic studies, showed the stabilization of the urinary tract and also the preservation of kidney function and not only was demonstrated in those patients with oral anticholinergic agent as their unique treatment, but also in those patients who had previously been derivated. The previous vesical stabilization allowed the optimum result of subsequent corrective surgeries in five patients (ureteroneocystostomy and vesiscotomy clousure. In cases with previous derivation, the treatment with oral anticholinergic agents made easier vesicostomic clousure in the forth quarter of the first year old wih the maintenance of the vesical stability. CONCLUSIONS: The use of oral anticholinergic agents at a dose of 0.2 mg/kg/day, has resulted to be safe as well as effective in the treatment of high risk bladder in infancy those with less than one year of life. In those patients with serious dysplasia by reflux, it contributes to the preservation of kidney function, it makes easier the early desderivation and it also stabilizes in a functional way the bladder as a previous step to subsequent reconstructive surgeries. PMID- 15901101 TI - [Results of laparoscopic treatment of complicated appendicitis]. AB - Laparoscopic appendectomy in children is a generally accepted procedure for the treatment of non-complicated acute appendicitis. Nevertheless, the role of laparoscopy in complicated cases is controversial. We show our experience with 40 cases of complicated acute appendicitis treated by means laparoscopy between February 2000 and October 2002. In every case we used 3 ports, one umbilical and the other two in both lower quadrants. The appendix was gangrenous in 31 patients and in the other 9 was perforated. Appendectomy was performed in an extracorporeal way in most of the cases. Average surgical time was 71 minutes and mean hospital stay was 8.8 days. Intraoperative complications occurred in 8 cases (20%) and postoperative complications were observed in 9 patients (22.5%). Four patients were reoperated (2 open and 2 laparoscopic) in order to drain two abscesses and treat two obstructions. The results of this serie is compared with another group of 40 complicated appendicitis operated in a classic open way in the same period of time. Postoperative complications were less often in the laparoscopic group. Oral intake, need for analgesia and hospital stay are more favorable in the laparoscopic group. PMID- 15901102 TI - [Reliability index of anorectal manometry for the diagnosis of Hirschspurng disease]. AB - The anorectal manometry (AM) and suction rectal biopsy (BS) are the main and accepted methods for diagnosis of Hirschsprung disease (HD). The reliability of these methods in newborns have been subject of controversy up to the present. MATERIAL AND METHODS: For diagnosis of HD purpose we have performed 2227 AM records in 1744 children. Initially we used a minichamber type probe joined to a close system to perform AM. From 1984 we developed a new type of 4 ways miniprobe connected to an open system which improved the reliability of the records. RESULTS: The indexes of reliability for our database have been the following: sensitivity 98%, specificity 97%, positive predictive value 85%, negative predictive value 99%, false positive 3.4% and false negative 1.8%. CONCLUSIONS: AM is a non invasive method with a high reliability, available in our hospital. It is a choice method for diagnosis of HD. PMID- 15901103 TI - [Importance of initial management in severe pediatric trauma]. AB - INTRODUCTION: Trauma is the most frequent cause of mortality in childhood and adolescence and causes almost 25% of admissions in Pediatric Intensive Care Units (PICU). We have evaluated the initial assesment of the severely injured children admitted in our PICU (pre-hospital care). MATERIAL AND METHODS: We reviewed the children younger than 16 years admitted in our PICU between January 1996 and December 2002. Prehospital caretakers, transportation after initial evaluation and therapeutic management were analized, using Pediatric Trauma Score (PTS) and Pediatric Risk of Mortality Score (PRISM) as predictors of injury severity and mortality, respectively. RESULTS: We treated 152 traumatized children in this period, 106 males and 46 females, with a mean age of 7.5 +/- 4.3 years. 116 patients received inmediate medical care with a mean PTS significatively greater than non-medical group (12 children). Non-medical caretakers treated 8.1% of severe trauma (PTS<8). Specialized transporter was inadequated in 7.1% of severe traumatized children. Gastric and vesical tube and spinal inmobilization were accomplished in 50%, specially in children with low PTS and high PRISM. We found a great variability in fluid and drugs administration. CONCLUSIONS: Although there has been a good evolution in treatment of pediatric trauma, in order to diminish morbidity and mortality it is necessary to identify and correct deficiencies in management, specially during the "golden hour", and train pre hospital caretakers in pediatric trauma management. PMID- 15901104 TI - [Biliary atresias operated with favourable results: predictable outcome]. AB - Since 1975, our experience in the treatment of biliary atresia with Kasai's technique has improved little by little, achieving 65% favourable outcome in the last five years. We define "good results" as the complete restoration of biliary flow and normalization of bilirrubin levels. The long-term evolution of these good results can be diverse. The objective of the present work is to analyze the outcome of patients in our series in whom a favourable initial response was achieved, as well as evaluating their present situation and future perspectives. The authors present a total of 17 patients operated by Kasai's technique since 1985, that constitutes the group with good results in our series. The controls were based on general analysis, liver function and periodic ultrasound explorations. All received a standardized medical treatment consisting of vitamin supplements (A, D3, E, K) minerals (zinc, calcium, phosphate, iron) ursodexoxicolic acid, luminal,as well as close control of calorie intake. In two patients the levels of bilirrubine were progressively increased with time, stabilizing at between 5/6 mgs/100 ml, with progressive hepatic hardening, appearance of splenomegalia, indirect signs of portal hypertension and a slight deterioration of hepatic function. One received a transplant at age 12 with Quick levels below 50%. The other, aged 16, continues with an acceptable hepatic function and good quality of life under recommendation of transplant. Eleven patients with ages ranging from fourteen months to seventeen years presented slight and firm hepatomegalia, moderate portal hypertension, GOT 71 +/- 8 mg/100 ml, GPT 97 +/- 11 mg/100 ml and normal bilirrubine levels. From this group, 3 patients, all under five years of age, experienced bleeding from esophageal varices which were controlled by sclerosis and medical treatment (propanolol and isosorbide dinitrate). Recently, one three year-old patient developed a hepatocarcinoma of rapid, mortal evolution. Since then, the determination of alfa feto protein in follow-up controls has been introduced. Four other patients of 5, 6, 14, 16, years of age are completely assymptomatic with an excellent clinical evolution. In our experience,the patients that overcome the third year after surgery without serious complications seem destined to reach puberty with a good quality of life. However, some cases show signs of hepatic fibrosis and portal hypertension, 77% in our series. Only 23% of patients with a favorable initial evolution appear to present a complete normalization of their hepatic lesion in the long term. PMID- 15901105 TI - [Chest wall distraction in thoracogenic scoliosis]. AB - INTRODUCTION: Combined congenital scoliosis and rib fusion associated with other chest deformities during infancy can lead to a progressive hypoplastic thorax that could be unable to support normal lung growth and respiratory function. Campbell introduced an expansion thoracoplasty technique in which fused ribs are separated and a vertical expandable prosthetic titanium rib is used as a chest wall distraction device to enlarge the affected hemithorax. This technique benefits the underlying lung by improving the thoracic volume and the respiratory function. PATIENTS AND METHODS: Four patients (3 boys and 1 girl) with severe unilateral thoracic deformity with combined scoliosis and rib fusion plus a restrictive respiratory insufficiency are presented herein; 1 of them needed continuous CPAP preoperatively. All had progressive scoliosis and failure to thrive. Preoperative evaluation included three-dimensional CT reconstruction. A thoracic expansion placing an intercostal vertical expandable prosthetic titanium device as a chest wall distractor was performed. Afterwards, distraction control was made every four months. RESULTS: Our 4 patients had a satisfactory outcome. Scoliosis was corrected and there were improvements of volume and function of the thorax. CONCLUSIONS: The placement of an intercostal distractor device improves the thoracic insufficiency syndrome, lengthening and expanding the thoracic cage at the same time. These effects benefit respiratory function and correct scoliosis, allowing an adecuate lung function. PMID- 15901106 TI - [Continent urinary diversion: the Mitrofanoff principle]. AB - INTRODUCTION: Continent urinary diversion (based on Mitrofanoff's principle), despite its complexity, is the gold standard in the treatment of those vesicourethral disfunctions that need clean intermittent catheterization (CIC) to achieve complete vesical voiding, in patients with non easily catheterizable native urethra. AIM: To analize our experience in continent urinary diversion at our centre. PATIENTS AND METHODS: We have reviewed the records of the 14 cases of continent urinary diversion in the last 8 years. They were grouped in order to the underliying condition: 1-Bladder exstrophy group (n=5): Mean age at diversion time was 5.5 years (range 3-7). In all patients the original Mitrofanoff s technique was performed (continent cutaneous appendicovesicostomy). Associated procedures were: ureterovesical reimplantation in all 5 cases; bladder neck reconstruction also in all 5 (3 of them needed vesicourethral transection); and bladder augmentation in two cases, using ileum and sigmoid respectively. 2- Myelomeningocele group (n=9, 10 procedures): Mean age was 11.5 years (range 6 16). Appendicovesicostomy was performed in 7 cases and a reconfigured ileum with the Casale technique was used in 3 cases (primarily in 2 and as an alternative in one). Associated procedures were: ureterovesical reimplantetion in 5 cases and bladder augmentation in 7 (using sigmoid in 4, ileum in 2 and urether in one case). RESULTS: In all patients complete continence was achieved. Complications found were: one appendicostomy prolapse, one appendix necrosis (that was then diverted with Casale's technique), one appendicular conduit stenosis and one case with catheterization difficulties that needed a tappering of the ileum conduit. Nowadays, 13 out of 14 pacients follow the CIC program each 3-4 hours without complication. CONCLUSION: Continent urinary diversion improves autonomy and life quality in those patients that need a definitive urinary diversion and have a long life expectancy. We have used these procedure with good results in patients with severe vesicourethral disfunction (of an intrinsic or neuropathic origin) in which clean intermittent catheterization was not possible through native urethra, or in patients with refractary incontinence in which vesicourethral transection was the only effective treatment. PMID- 15901107 TI - [Acute secundary abdomen to pancreatic hydatid cyst]. AB - The hydatidosis is a frequent zoonosis in Spain, but isolated location in pancreas and their onset as acute abdomen is excepcional. We present a boy 14 years old, with abdominal pain and low-grade fever since 1 month. Hemogram shows eosinophilia, and echography an anechoic mass in tail of pancreas beside of free peritoneal liquid. At surgery procedure: a broken hydatid cyst is verified, surrounded of tissues inflammatory reaction that evolved to pancreatic fistula. The differents forms from clinical presentation of the pancreatic hydatid cyst, as well as the complications related to their location are analyzed. PMID- 15901108 TI - [Congenital cystic adenomatoid malformation associated with pulmonary sequestration]. AB - The association of congenital cystic adenomatoid malformation with pulmonary sequestration is infrequent. Has been reported less than 60 cases. We describe two patients with this association. One of this with extralobar pulmonary sequestration, and the other with an intralobar one. PMID- 15901109 TI - [Congenital colonic atresia. Surgery correction in one or two stages?]. AB - Sigmoid atresia is rare, the recognzed prevalence is 1 per 15.000 to 60.000 live born. We present a 30 hours old boy with vomiting, abdominal distension and failure to pass meconium. Physical examination showed severe abdominal distension. An enema was performed and a gray mucus plug taken out. The abdominal plain XR showed bowel obstruction and in a barium enema only 5-7 cm of microcolon was seen. With the diagnosis of sigmoid atresia the patient underwent surgery, observing one atresia type I, with dilated proximal loop and distal microcolon. A colostomy was performed and few days later one end-to-end anastomosis was made. PMID- 15901110 TI - [Torsion of paraovarian cyst in pediatrics]. AB - The paraovarian cyst ploughs to little frequency in pediatric population. We report a case of twist proved at surgery, and the difficulty of preoperatory diagnosis. It is very important to consider the possibility of twist to be a complication of a paraovarian cyst to try to prevent salpinguectomy. PMID- 15901111 TI - [Free fat flap for the treatment of Parry-Romberg disease in children]. AB - Progressive hemifacial Atrophy (PHA) or Romberg's Disease is a rare entity of unknown etiology, which most often affects only one side of the face and can include all tissue planes in different grades of severity. Of the several treatments proposed, pedicled or free flaps seem to achieve the best cosmetic results in severe cases. In mild to moderate cases, free autologous fat transplant may obtain optimal results with lower complexity, cost and morbidity. The presence of multiple adipoblasts might increase the possibilities of graft take, particularly in children. Furthermore, refinements in the processes of extraction, preparation and reintroduction of the fat have contributed to improve the results that may be long lasting. We present 2 cases of PHA in 2 girls (11 and 13 years old), that were treated by lipofilling. No complications occurred. In both cases we observed a small reduction in the volume of the initial graft. Thus the procedure was repeated 6 months later to improve facial symmetry. Results have been excellent in one year follow-up. PMID- 15901112 TI - Testimony as an expert witness--medical ethics and professionalism count. PMID- 15901113 TI - You've got mail. PMID- 15901114 TI - Colorectal cancer screening by primary care physicians in Iowa. PMID- 15901115 TI - Cranial vault modification as a cultural artifact: a comparison of the Eurasian steppes and the Andes. AB - This paper details the practice of intentional cranial vault modification in the Eurasian steppes as well as in the pre-Columbian Andes focusing on the similarities and differences in how the practice was used to respond to changes in society. The appearance of vault modification in the steppes and the forms seen in the cemeteries of the Syr Darya and Amu Darya River deltas are discussed. Temporal changes in the pattern of modification are also investigated, especially the dramatic homogenization of the custom resulting from the conquests of the Huns. This is contrasted with incidences of cranial modification in the south central Andes, including the appearance of deliberate head shaping as well as shifts in the practice during the expansion of the Bolivian Altiplano state of Tiwanaku. Similarities in the use of cranial vault modification between these unrelated areas and in the alterations of the practice resulting from foreign contact are considered in light of vault modification's role as a malleable cultural artifact. PMID- 15901116 TI - Three-dimensional technology for linear morphological studies: a re-examination of cranial variation in four southern African indigenous populations. AB - In order to compare linear dimensions made by traditional anthropometric techniques, and those obtained from three-dimensional coordinates, samples of four indigenous southern African populations were analysed. Linear measurements were obtained using mathematically transformed, three-dimensional landmark data on 207 male crania of Cape Nguni, Natal Nguni, Sotho and Shangaan. Univariate comparisons for accuracy of the transformed linear data were made with those in a traditional linear study by de Villiers (The Skull of the South African Negro: A Biometrical and Morphological Study. Witwatersrand University Press, Johannesburg) on similar samples and equivalent landmarks. Comparisons were not made with her Penrose (Ann Eugenics 18 (1954) 337) analysis as an apparently anomalous 'shape'-'size' statistic was found. The univariate comparisons demonstrated that accurate linear measurements could be derived from three dimensional data, showing that it is possible to simultaneously obtain data for three-dimensional geometric 'shape' and linear interlandmark analyses. Using Penrose and canonical variates analyses of the transformed three-dimensional interlandmark measurements, similar population distances were found for the four indigenous southern African populations. The inter-population distance relationships took the form of three separated pairs of distances, with the within-pair distances very similar in size. The cranial features of the four populations were found to be overall very similar morphometrically. However the populations were each shown by CVA to have population specific features, and using discriminant analyses 50% or more of the individual crania (with the exception of the Sotho) could be referred to their correct populations. PMID- 15901117 TI - How close to a pendulum is human upper limb movement during walking? AB - The aim of this work was to investigate how close to pendulum-like behaviour the periodic motion of the human upper limb (or upper extremity) is, during normal walking at a comfortable speed of locomotion. Twenty-five healthy young persons (males and females) participated in the experiment. Biomechanical testing was undertaken (mass and centre of mass of each segment of the total upper extremity). Participants were walking on a treadmill with a standardised velocity of 1.1 ms(-1) (comfortable speed for all of them). A video analysis system with Silicon software was used to measure the different angles of the arm and forearm. The theoretical period of motion and maximal angular velocity were computed for the centre of mass of the total upper limb from the measured phases of the arm swing and associated positional potential energies. Actual measured periods of motion, in comparison, represented a level of similarity to a lightly damped simple pendulum. Using this assumption, the "damping factor" was calculated from the ratio between theoretical and measured values. A vast majority of people exhibited an actual angular velocity exceeding the expected theoretical angular velocity calculated for a virtual pendulum of similar mass and length characteristics. This may be due to muscle forces that are contributing to the motion of the upper limb during walking rather than simple gravity force acting alone. The observed positional potential energy of the dominant limb was greater than that of the non-dominant limb for the vast majority of participants. PMID- 15901119 TI - Patella measurements of South African whites as sex assessors. AB - This study uses metrical characteristics of the patella to derive population specific equations for sex determination in South Africa. Six measurements were taken from 120 normal and undamaged left patellae in a sample of known race, equally distributed by sex, obtained from the Raymond A Dart collection of human skeletons. These data were subjected to discriminant analysis. Maximum height and maximum breadth were selected in the stepwise analysis with an average accuracy of 83% in correct sex classification. Four functions were derived from the direct analysis with a range of average accuracy between 77% and 85%. While most individual variables showed high misclassification rates and may not be useful on their own, maximum height and maximum breadth have an average accuracy of 85 and 79%, respectively. PMID- 15901118 TI - Pattern influence on the fingers. AB - In this study dermatoglyphic traits in 16 populations in northern Hungary were examined. The occurrence and probability of the manifestation of finger pattern types were analysed. Certain relationships were found that show that if a given pattern type is present on one of the fingers, then, in these cases, the frequency of pattern types on the other nine fingers typically is different from that in the entire sample. We termed this relationship or effect; pattern influence. In most cases there was a strong significant difference in frequency values between the entire sample and the studied cases. We studied the relationship of populations and pattern types using correspondence analysis. In the entire sample, most populations are located around the ulnar loop and the whorl. This arrangement is significantly and typically different in pattern influence cases. Most populations aggregated around the pattern type in which pattern influence was studied. Pattern influence was found in all populations studied, and in both sexes. PMID- 15901120 TI - Lateral epicondylalgia or epicondylitis: what's in a name? PMID- 15901121 TI - The relationship between lumbar segmental motion and pain response produced by a posterior-to-anterior force in persons with nonspecific low back pain. AB - STUDY DESIGN: Cross-sectional. OBJECTIVE: To investigate the association between lumbar segmental motion and pain response during the application of a posterior to-anterior (PA) force to the lumbar spinous processes in persons with nonspecific low back pain. BACKGROUND: Although low back pain is believed to be associated with altered segmental motion of the lumbar spine, the relationship between subjective reports of pain and objective measurements of segmental motion has not been established. METHODS AND MEASURES: Thirty-five individuals between 18 and 45 years of age with nonspecific low back pain (less than 3 months' duration) participated. All subjects participated in 2 separate procedures: (1) segmental motion assessment during a PA force application over the lumbar spinous processes using dynamic magnetic resonance imaging (MRI), and (2) pain assessment during a PA force application procedure outside of the MRI environment. Frequency counts were used to determine the lumbar segments that were most painful, and which functional spinal units had the most and least motion. Fisher exact tests were performed to determine if an association existed between the most painful segment and the functional spinal unit with the most or least motion. RESULTS: L5 was deemed the most painful segment in nearly half of the participants (48.1%). The L1-2 and L3-4 functional spinal units most frequently had the most motion (25.9% each) and the L4-5 functional spinal units most frequently had the least motion (29.6%). No association was found between the most painful segment and the functional spinal units with either the most or least motion. CONCLUSION: The results of this study indicate that an assumption regarding segmental motion cannot be inferred from the pain response when using a PA force application procedure. PMID- 15901122 TI - The addition of the Protonics brace system to a rehabilitation protocol to address patellofemoral joint syndrome. AB - STUDY DESIGN: Randomized clinical trial. OBJECTIVES: To investigate the clinical efficacy of the addition of the Protonics system to a standard exercise-based patellofemoral rehabilitation protocol. BACKGROUND: The Protonics system has been suggested as an intervention for patients with patellofemoral pain syndrome (PFPS). However, the effects of this system have not been compared to the effects associated with traditional exercise-based rehabilitation alone. METHODS AND MEASURES: Seventeen of 34 females (mean age, 28 years; range, 13-55 years) diagnosed with PFPS were randomly assigned to wear the Protonics system while participating in a conventional exercise-based rehabilitation program. Functional and patient-reported outcome measures were evaluated, including Kujala score and the lateral step-up test. In addition, measurements of hip internal and external rotation, hip extension, and iliotibial band muscle length were compared between groups. RESULTS: Patients in both groups demonstrated improvement in Kujala score (P<.001), performance on the lateral step-up test (P<.001), and pain during the step-up test (P<.001) at the conclusion of the study. However, there was no difference between groups with respect to improvement in Kujala score (P = .33), step-up test performance (P = .47), or pain during the step-up test (P = .24). Patients using the Protonics system demonstrated greater gain in passive hip extension (P = .023) and increased hip external rotation motion (P = .017) at discharge versus patients treated with exercise alone. However, there was no difference in iliotibial band flexibility (P = .80) or hip internal rotation motion (P = .09) between groups. A greater proportion of patients in the Protonics group reported no pain with step-up testing at each 2-week interval. However, the 2.2 fewer visits required by patients in the Protonics group to meet discharge criteria did not achieve statistical significance (P = .08). CONCLUSIONS: Patients using the Protonics system demonstrated a shift in available hip rotation and increased passive hip extension flexibility. However, these changes were not outside the bounds of potential measurement error and did not translate into significant functional differences from a similar group treated with exercise alone. The economic implications of an average 2.2-visit decrease in treatment sessions per patient using the Protonics system are uncertain. PMID- 15901123 TI - Disability and walking capacity in patients with lumbar spinal stenosis: association with sensorimotor function, balance, and functional performance. AB - STUDY DESIGN: One-group, prospective, cross-sectional study. OBJECTIVES: To determine how sensorimotor function, balance, and physical performance are associated with disability and walking capacity in patients with lumbar spinal stenosis. BACKGROUND: Disability and limited walking capacity are often reported by patients with lumbar spinal stenosis. Identification of associated factors could provide information for future investigations leading to better prevention and intervention strategies. METHODS AND MEASURES: Fifty patients with lumbar spinal stenosis answered questions regarding symptom intensity, disability, and walking capacity. Muscle strength and vibration sense were assessed to represent sensorimotor function. Balance ability was measured by single-leg stance time and basic physical performance was tested by the up-and-go (UG) test. Regression analyses, entering demographics and symptom intensity as control variables, and sensory, strength, balance, and physical performance as additional independent variables, were conducted separately for disability and walking capacity. RESULTS: Symptom intensity, vibration sense at the big toe, and UG test time were significantly correlated with disability. The final regression model showed that the control variables explained 20% of the variance, while vibration sense and UG test time explained an additional 20% of the variance. Walking capacity was significantly correlated with vibration sense at the big toe and UG test time. No significant regression model emerged for walking capacity. CONCLUSIONS: A moderate amount of variance in disability could be explained by sensory function at the big toe and physical performance. These factors should be considered in future research. PMID- 15901124 TI - The effect of long versus short pectoralis minor resting length on scapular kinematics in healthy individuals. AB - STUDY DESIGN: Two-group comparison. OBJECTIVE: To compare scapular kinematics during arm elevation between groups distinguished by pectoralis minor resting length. BACKGROUND: Studies have demonstrated that individuals with subacromial impingement have altered scapular kinematics, such as loss of posterior tipping and increased internal rotation. One proposed mechanism for these alterations is an adaptively short pectoralis minor. This anterior scapulothoracic muscle may impact normal scapular kinematics if adaptively short. METHODS AND MEASURES: Fifty volunteers without shoulder pain were divided into long or short groups according to normalized pectoralis minor resting length. An electromagnetic motion capture system determined the angular orientation of the scapula, humerus, and trunk during arm elevation in 3 separate planes. Groups were compared for 3 dimensional scapular orientation relative to the trunk at arm elevation angles of 30 degrees, 60 degrees, 90 degrees, and 120 degrees, using a mixed-model analysis of variance (ANOVA). RESULTS: There were statistically significant interaction effects between group and arm elevation angle for scapular tipping in all planes of arm elevation, with the scapula for the short group staying anteriorly tipped at higher angles. There was also a significant interaction for scapular internal rotation at lower arm elevation angles in the coronal plane only, with individuals with a shorter pectoralis minor demonstrating a more internally rotated scapula. CONCLUSIONS: The group distinguished by a short pectoralis minor demonstrated scapular kinematics similar to the kinematics exhibited in earlier studies by subjects with shoulder impingement. These results support the theory that an adaptively short pectoralis minor may influence scapular kinematics and is therefore a potential mechanism for subacromial impingement. PMID- 15901125 TI - Torque responses in human quadriceps to burst-modulated alternating current at 3 carrier frequencies. AB - STUDY DESIGN: Independent group design, 1 group per carrier frequency, with subjects stratified by gender and systematically assigned to groups. OBJECTIVE: To determine the effect of the frequency of burst-modulated alternating current (AC) on the electrically induced torque (EIT) response of the quadriceps femoris muscle. BACKGROUND: Many studies have examined the influence of current amplitude and burst modulation on the EIT response. However, research is limited on the effects of carrier frequency on the EIT response of the quadriceps femoris. METHODS AND MEASURES: Twenty-three healthy subjects (mean age +/- SD, 22.7 +/- 1.7 years) volunteered for the study and were systematically assigned to 1 of 3 groups: group 1 (2500-Hz AC), group 2 (3750-Hz AC), and group 3 (5000-Hz AC). For normalization purposes, maximal voluntary isometric contraction (MVIC) force was assessed with a load cell prior to the application of 5 minutes of neuromuscular electrical stimulation (NMES). The EIT and current amplitude were recorded during each of the 5 minutes of NMES. RESULTS: The mean EIT ([EIT/MVIC] x 100) produced at equivalent current amplitudes when using the 2500-Hz carrier frequency was significantly greater than the mean EITs generated at 3750 Hz (P = .004) and 5000 Hz (P<.001), respectively. CONCLUSIONS: Burst-modulated alternating current delivered at 2500 Hz produced greater EITs than those generated at 3750 and 5000 Hz. PMID- 15901126 TI - The influence of gender and age on hamstring muscle length in healthy adults. AB - STUDY DESIGN: Cross-sectional descriptive study. OBJECTIVES: To examine the factors of gender and age, stratified by 10-year increments, on hamstring muscle length (HML) as measured by passive straight-leg raise (PSLR) and popliteal angle (PA). BACKGROUND: Differences in HML between men and women have not been examined for a large group of healthy adults over a wide range of ages. The usefulness of these data is to provide some typical values of HML for future reference. METHODS AND MEASURES: Two hundred fourteen adults (108 women, 106 men; age range, 20-79 years) with no known history of hip or knee joint disease and no history of recent hamstring strain participated in the study. PSLR (trunk-thigh angle) and PA (thigh-leg angle) were estimated with a goniometer. A 2-way analysis of variance (ANOVA) was used to analyze the effects of 2 independent variables (gender and age) on 2 dependent variables (PSLR and PA). Statistical significance was established at alpha<.05. RESULTS: HML differed significantly (P<.001) between genders for both methods of measurement, with females demonstrating greater flexibility than their male counterparts. The difference between genders was 8 degrees for PSLR and 11 degrees for PA. HML was not influenced by age. CONCLUSIONS: This study provides physical therapists with typical values of HML in healthy men and women. PMID- 15901127 TI - Immunohistochemical profile and c-kit mutations in gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) are low-grade sarcomas arising from the interstitial cells of Cajal, harboring mutation of c-kit. We investigated the morphological, immunohistochemical, and molecular profile of 55 GISTs to establish the prevalence of mutations, their clinical significance, and diagnostic utility. c-kit mutations were investigated by evaluating the entire coding sequence of the gene with non-radioisotopic PCR-SSCP, and characterized with fluorescent cycle sequencing. Mutations were detected in 39 tumors (71%), the majority (67%) involving exon 11. Two tumors showed exon 9 mutations (one tumor located in the small intestine and one in the stomach), whereas two cases showed a polymorphism at the splicing site of exon/intron 1 present in healthy blood donors with a 3% frequency. CD117 was expressed in 53 tumors (96%); CD34 was positive in 42 cases (76%); 42 cases (76%) expressed both CD117 and CD34. c kit mutations were similarly distributed in stromal tumors at low risk of aggressive behavior (78%), intermediate risk (66%), and high risk (71%). Fifteen tumors expressing CD117 showed wild-type kit gene, and on histological grounds, they were equally distributed among epithelioid and spindle cell morphology. One case neither expressed CD117 nor did it show c-kit mutation. Data suggest that both immunohistochemical and molecular evaluation may be useful in tumors likely to be classified as GISTs; molecular analysis appears valuable to support the diagnosis and to identify cases that can benefit from recent novel therapeutic tools. PMID- 15901128 TI - Overexpression of CD10 and reduced MUC2 expression correlate with the development and progression of colorectal neoplasms. AB - There are two opposing theories of the natural history of colorectal neoplasm, adenoma-carcinoma sequence and de novo carcinogenesis. To elucidate the histogenesis of colorectal carcinoma, we investigated the expression of CD10, MUC2, MUC5AC, MUC6, and p53 in colorectal neoplasms. Sixty-seven morphologically distinct neoplastic specimens were divided into the following groups according to morphology: adenoma (groups A and B), protruded-type carcinoma (group C), superficial-type carcinoma with adenomatous component (group D), or superficial type carcinomas without any adenomatous component (group E). Diagnoses of adenomas and carcinomas were based upon the Vienna classification of gastrointestinal epithelial neoplasia. The expression of CD10 in group E lesions was more intense than in the other groups. Regardless of morphology, MUC2 expression was significantly decreased in CD10-positive carcinomas, and the p53 positive rate was much higher in CD10-positive than in CD10-negative carcinomas. The overexpression of CD10 and reduced expression of MUC2 may be associated with the development and progression of colorectal carcinoma. A specific tendency was evident in superficial-type carcinomas without any adenomatous component (de novo carcinomas). These carcinomas are considered to be more aggressive than other morphologically distinct carcinomas. PMID- 15901129 TI - VEGFR-3 expression in breast cancer tissue is not restricted to lymphatic vessels. AB - We examined the immunohistochemical reactivity for vascular endothelial growth factor receptor 3 (VEGFR-3), a protein playing an important role in lymphangiogenesis, in breast cancer. A retrospective series of 77 invasive ductal breast carcinomas was investigated. The relationship between VEGFR-3 expression and clinicopathologic parameters was examined for statistical significance using Pearson's chi-square (chi2) test and Fisher's exact test (when n<5). Threshold for significance was p<0.05. Patient age ranged from 31 to 77 years (mean: 55 years). The VEGFR-3 immunoreactivity was as follows: 5 cases were negative (6.5%), 35 + (45.4%), 27+ + (35.1%), and 10+ + + (13.0%). Reactions were positive for both lymphatic and blood vessels in several cases. VEGFR-3-positive reactions were more frequent in the tumor periphery than within the tumor. Immunoreactivity was also observed in myoepithelial cells surrounding both normal ducts and ducts with ductal carcinoma in situ. Statistical analysis of VEGFR-3 reactions was not significantly related to node status, microvessel density, and tumor grade. Ploidy showed a tendency towards significance (p=0.063); however, owing to the limited number of cases, statistical significance was not reached. VEGFR-3 lacks lymphatic vessel specificity and is also expressed in blood vessels, myoepithelial cells, and neoplastic cells. PMID- 15901130 TI - HMGA1 proteins in human atherosclerotic plaques. AB - One of the major characteristics of atherosclerosis is the migration of smooth muscle cells (SMC) from the tunica media to the intima, caused by alterations in the environment, e.g. mechanical, chemical, or immunologic injuries of the arterial walls. A group of molecules that may act as a main regulator of SMC phenotype switching is formed by the so-called HMGA1 high-mobility group proteins. One target gene of the HMGA1 protein, playing a major role in the development of atherosclerotic lesions, is CD44. The expression of CD44 is regulated by IL-1beta, but binding of HMGA1 potentiates the transactivation of the CD44 promoter. In this study, the HMGA1 expression of human atherosclerotic plaque samples was examined. Compared to the non-active components, all major components of the well-developed atherosclerotic plaques showed strong positivity of the high-mobility group protein HMGA1 in their activated areas, e.g. neointimal SMCs, macrophages, newly built blood vessels. This report is the first to describe HMGA1 as one of the first mediators in the development of human atherosclerotic plaques. PMID- 15901131 TI - Correlation between DNA alterations and p53 and p16 protein expression in cancer cell lines. AB - To investigate the interaction between DNA abnormalities, p53 and p16 gene mutations, and methylation and protein expression, 20 cancer cell lines were examined by Western blotting. A clear relation was found to exist between p53 accumulation and mutation status. Of 20 cell lines examined, 14 demonstrated p53 homozygous mutations in exons 3-10, including 12 missense mutations, one nonsense mutation, and one frameshift mutation. Overexpression of p53 was always linked to missense mutations in exons 6-8. Intermediate expression of p53 was noted in cells with missense mutations or polymorphism to proline at codon 72 in exons 4 5, whereas there was slight or no visible expression in wild type cells and in cells with nonsense and frameshift mutations. DNA aberration in the p16 promoter gene correlated significantly with protein expression of the p16 suppressor gene. Overexpression was noted in six cell lines, intermediate expression in two, and slight or no visible expression in 12. Methylation-caused disappearance of p16 protein was noted in 40% (8/20) of the cell lines. Of six cell lines overexpressing p16 protein, two could be amplified with primers for both unmethylated and methylated forms in a methylation-specific RCP analysis. One cell line with no visible expression could also be amplified with both primers. Overexpression or disappearance of p16 protein may readily occur when one of two alleles has been methylated. PMID- 15901132 TI - Are external quality assurance (EQA) slide schemes a valid tool for the performance assessment of histopathologists? AB - Quality assurance plays a vital role in the healthcare profession and histopathologists play a central role in the diagnosis and treatment of disease. In the past these specialists have worked in isolation and quality assurance of their work has been difficult. In recent years this has changed with the introduction of External Quality Assurance slide schemes. This paper discusses how these schemes have evolved, the problems of standard setting and their validity as a measure of pathologists performance. PMID- 15901133 TI - Generalized herpes simplex virus infection in an immunocompromised patient- report of a case and review of the literature. AB - Patients with immunodeficiency or treatment-related immunosuppression are at an increased risk of developing severe herpes simplex virus (HSV) infection. We present a fatal case of a generalized HSV-1 infection in a 22-year-old female afflicted by acute lymphoblastic leukemia who was treated with polychemotherapy. The terminal clinical course was characterized by abdominal pain, progressive hepatic failure, and disseminated intravascular coagulation. Autopsy revealed non perioral herpetic skin lesions and mucosal ulceration of the esophagus and colon. Punctuated areas of yellow-tan necrosis with hyperemic rims were detected in the liver, spleen, and lung. Numerous petechiae were observed on the mucosal surface of the esophagus, jejunum, ileum, and colon. Microscopically, lesions demonstrated the cellular changes characteristic of herpetic infection. Immunohistochemistry for identification of the virus using monoclonal antibodies against HSV-1 and HSV-2 showed positive staining for HSV-1. Polymerase chain reaction and sequencing confirmed HSV-1 positivity. Emphasis must be placed on clinical awareness of a generalized HSV infection in immunocompromised patients. Absence of orofacial or genital lesions does not rule out the possibility of active HSV infection. PMID- 15901134 TI - Low-grade central osteosarcoma of the skull. AB - A case of low-grade central osteosarcoma of the skull in a 36-year-old male is described. A 10cm mass of the left temporo-occipital bone, which was incidentally found 6 years previously on the occasion of a head trauma, was osteolytic and osteoplastic, affecting soft tissues and the subdural space. The patient was treated with intralesional curettage. Histologically, the tumor was characterized by spindle cell proliferation with woven and lamellar bone formation, resembling fibrous dysplasia, parosteal osteosarcoma, low-grade fibrosarcoma, or desmoplastic fibroma. The spindle cells were fairly uniform with mild atypia and low-mitotic activity (3 per 50 high power fields), and cellularity varied from low to high. Clinicians and pathologists should be aware of this type of low grade osteosarcoma of the skull and distinguish it from its mimics, including fibrous dysplasia, parosteal osteosarcoma, and low-grade fibrosarcoma. Careful microscopic correlation with radiographic findings is the clue to the correct diagnosis. PMID- 15901135 TI - Mucinous adenocarcinoma of posterior urethra. Report of a case. AB - Primary carcinoma of the male urethra accounts for less than 1% of malignancies in males. Mucinous adenocarcinoma of the urethra is extremely rare, and its biologic behavior is not well known. We report a case of mucinous adenocarcinoma showing the histologic features of colloid adenocarcinoma that appears to have evolved either by neoplastic degeneration of goblet cells found in the urethral epithelium or by malignant degeneration of persistent glandular elements of uretheritis cystica and glandularis. PMID- 15901137 TI - A case of vulvar superficial angiomyxoma with necrotizing angiitis-like lesions and expression of granulocyte-colony stimulating factor. AB - A vulvar tumor of 3-year-old girl was resected. The tumor had a pedunculated polypoid appearance with a multinodular surface and was covered by normal colored skin. Histologically, the tumor was lobulated and consisted of sparse stellate- or spindle-shaped tumor cells with a large amount of edematous stroma admixed with myxomatous areas. The tumor was rich in blood vessels of various sizes. Several blood vessels showed fibrinoid necrosis. There was a diffuse neutrophilic infiltration in the stroma. The tumor was diagnosed as a superficial angiomyxoma. Immunohistochemically, the tumor cells diffusely expressed vimentin, focally alpha-smooth muscle actin and desmin. Both estrogen and progesterone receptors were negative. Occasionally, they expressed CD34. Most of the tumor cells expressed granulocyte-colony stimulating factor (G-CSF). Endothelial cells of tumor blood vessels occasionally expressed intercellular adhesion molecule-1 or E selectin. Some endothelial cells in the tumor were immunolabeled by anti-vascular endothelial growth factor (VEGF) antibody along their luminal surfaces. In the present case, G-CSF and adhesion molecules to neutrophils may have played some roles in neutrophilic infiltration into the tumor and in fibrinoid necrosis of the blood vessels. In addition to these molecules, VEGF may have contributed to vascular growth, leading to edematous stroma. PMID- 15901136 TI - HER-2/neu oncogene in uterine carcinosarcoma on tamoxifen therapy. AB - HER-2/neu is an oncogene located on chromosome 17, encoding a type 1 tyrosine kinase growth factor receptor. HER-2/neu is overexpressed in 25-30% of breast cancers, increasing the aggressiveness of the tumor. We describe HER-2/neu overexpression and her-2/neu oncogene amplification in a case of uterine carcinosarcoma occurring in a 46-year-old women who had undergone mastectomy and a 2-year postoperative treatment with tamoxifen for invasive breast cancer. This is the first study demonstrating HER-2/neu expression and her-2/neu oncogene amplification in a uterine carcinosarcoma that has developed in a patient given tamoxifen therapy. It still needs to be clarified whether HER-2/neu overexpression increases the aggressiveness of carcinosarcoma, or whether HER 2/neu has a direct role in its pathogenesis, as described in breast cancers. Our observation of the her-2/neu oncogene amplification does not shed light on the prognostic impact of uterine carcinosarcoma following tamoxifen therapy, but it may indicate the need for further studies of HER-2/neu overexpression in a larger series of uterine carcinosarcoma patients, and it may permit us to hypothesize about a therapeutic concept, including the inhibition of HER-2/neu by humanized monoclonal antibodies also in uterine carcinosarcoma patients. PMID- 15901138 TI - Withdrawal of immunosuppression contributing to the remission of malignant melanoma: a case report. PMID- 15901139 TI - Synthesis of C3-symmetric tris(beta-hydroxy amide) ligands and their Ti(IV) complex-catalyzed enantioselective alkynylation of aldehydes. AB - [reaction: see text]. A series of new chiral C3-symmetric tris(beta-hydroxy amide) ligands have been synthesized via the reaction of 1,3,5 benzenetricarboxylic chloride and optically pure amino alcohols (up to 96% yield). The asymmetric catalytic alkynylation of aldehydes with these new C3 symmetric chiral tris(beta-hydroxy amide) ligands and Ti (O(i)'Pr)4 was investigated. Ligand 4c synthesized from (1R,2S)-(-)-2-amino-1,2-diphenylethanol is effective for the enantioselective alkynylation of various aldehydes, and high enantioselectivity was obtained with aromatic aldehydes and alpha,beta unsaturated aldehyde (up to 92% ee). PMID- 15901140 TI - Superparamagnetic nanoparticle-supported catalysis of Suzuki cross-coupling reactions. AB - Emulsion polymerization was examined as a novel route for the synthesis of core/shell superparamagnetic nanoparticles consisting of a highly crystalline gamma-Fe2O3 core and a very thin polymeric shell wall. These nanoparticles were used as soluble supports for immobilizing Pd catalysts to promote Suzuki cross coupling reactions. Recovery of catalysts was facilely achieved by applying a permanent magnet externally. Isolated catalysts were reused for new rounds of reactions without significant loss of their catalytic activity. PMID- 15901141 TI - Stereoselective photooxidation of enecarbamates: reactivity of ozone vs singlet oxygen. AB - [reaction: see text]. Oxazolidinone-functionalized enecarbamates show contrasting behavior upon oxidation by singlet oxygen and by ozone. The observed stereoselectivity difference indicates that the oxidation with ozone is subject to classic steric effects, whereas the very high selectivity in the photooxidation with singlet oxygen is derived from vibrational deactivation. PMID- 15901142 TI - Solvent effect on concertedness of the transition state in the hydrolysis of p nitrophenyl acetate. AB - [reaction: see text]. The reaction pathway for the alkaline hydrolysis of p nitrophenyl acetate is investigated using density functional theory. It is shown that solvent plays an indispensable role in shaping the concerted transition state. The concertedness of this transition state is supported by good agreement with the measured kinetic isotope effects. PMID- 15901143 TI - Ruthenium-catalyzed diyne hydrative cyclization: synthesis of substituted 1,3 diene synthons. AB - [reaction: see text]. A novel and versatile strategy for the synthesis of highly functionalized substituted 3-sulfolenes based on [CpRu(CH3CN)3]PF6-catalyzed hydrative cyclization has been developed. A marked ketone directing effect in ruthenium-catalyzed cyclization was observed for the first time. This provides complementary chemoselectivity for the synthesis of 3-sulfolenes and other cyclic enones. The utility of this method has been demonstrated by SO2 extrusion of 3 sulfolenes to afford 1,3-dienes and the subsequent inter- and intramolecular Diels-Alder reaction. PMID- 15901144 TI - Suzuki coupling of aryl chlorides with phenylboronic acid in water, using microwave heating with simultaneous cooling. AB - [reaction: see text]. We present here a methodology for the Suzuki coupling of aryl chlorides with phenylboronic acid using Pd/C as a catalyst, water as a solvent, and microwave heating. We show that simultaneous cooling in conjunction with microwave heating prolongs the lifetime of the aryl chloride substrates during the course of the reaction and, as a result, yields of the desired biaryl as well as overall recovery of material can be increased. PMID- 15901145 TI - Titanium-mediated alkylative coupling of N-acylpyrroles. AB - [reaction: see text]. Inter- and intramolecular titanium-mediated coupling reactions of N-acylpyrroles are reported for convenient functionalization of terminal olefins. Comparison with the Kulinkovich reaction of esters and other carboxylic acid derivatives is also included. PMID- 15901146 TI - Directed library of anilinogeranyl analogues of farnesyl diphosphate via mixed solid- and solution-phase synthesis. AB - [reaction: see text]. A directed library of anilinogeranyl diphosphate analogues of the isoprenoid farnesyl diphosphate has been prepared by solid-phase organic synthesis using a traceless linker strategy in moderate yield in three steps: reductive amination, bromination, and treatment with ((n-Bu)4N)3HP2O7. PMID- 15901147 TI - Synthesis of unsaturated amino alcohols through unexpectedly selective Ru catalyzed cross-metathesis reactions. AB - [reaction: see text]. A two-step synthesis of N-protected unsaturated amino alcohols is disclosed that relies on an unexpectedly selective cross-metathesis (CM) involving allyl cyanide and pent-4-en-1-ol. The solution concentration and the identity of the Ru complex used are critical to the selectivity and efficiency of CM reactions. The intermediate obtained by CM is converted efficiently to the final desired products through a one-pot nitrile reduction/amine protection procedure. PMID- 15901148 TI - Synthesis of allenamides by copper-catalyzed coupling of allenyl halides with amides, carbamates, and ureas. AB - [reaction: see text]. A variety of N-substituted allenes have been synthesized by the copper-catalyzed coupling reaction between allenyl halides and amides, carbamates, and ureas. The reactions proceed in good to excellent yield using 7 mol % copper thiophenecarboxylate and 15 mol % of a diamine catalyst. PMID- 15901149 TI - Reaction of diaryl-1,2-diones with triphenylphosphine and diethyl azodicarboxylate leading to N,N-dicarboethoxy monohydrazones via a novel rearrangement. AB - [reaction: see text]. A mechanistically novel reaction of diaryl-1,2-diones with diethyl azodicarboxylate and triphenylphosphine to afford N,N-dicarboethoxy monohydrazones is described. The reaction proceeds via a nitrogen to nitrogen migration of a carboethoxy group. PMID- 15901150 TI - Unexpected reaction of dimethyl acetylenedicarboxylate with in situ generated arylketenes catalyzed by 1-methylimidazole. AB - [reaction: see text]. An unexpected 1-methylimidazole-catalyzed reaction of dimethyl acetylenedicarboxylate (DMAD) with in situ generated arylketenes leading to the synthesis of dimethyl 2-arylidenesuccinates under mild conditions is described. A plausible mechanism has been proposed. PMID- 15901152 TI - New fluorescent chemosensor based on exciplex signaling mechanism. AB - [reaction: see text]. A novel fluorescent chemosensor (compound 1) containing aminonaphthol, which selectively recognizes fluoride anion with high sensitivity, was synthesized. The fluorescence of compound 1 was quenched rapidly by fluoride ion, and a new peak at a longer wavelength emerged concurrently, which constituted the signature for fluoride detection. The mechanism of exciplex formation was proposed for the interesting observation. PMID- 15901151 TI - Magic ring catenation by olefin metathesis. AB - [reaction: see text]. Olefin metathesis has been employed in the efficient syntheses of a [2]catenane with the templation being provided by the recognition between a secondary ammonium ion and a crown ether. In one approach, a crown ether precursor has been clipped around an NH2+ center situated in a macrocyclic ring, yielding the mechanically interlocked compound. In the other approach, the reversible nature of olefin metathesis allows for a magic ring synthesis to occur wherein two free macrocycles can be employed as the stationary materials, leading to the formation of the same [2]catenane. PMID- 15901153 TI - Ruthenium-catalyzed decarboxylative insertion of electrophiles. AB - [reaction: see text]. A ruthenium complex, Cp*Ru(bipyridyl)Cl, has been developed as a catalyst for the first regioselective tandem Michael addition-allylic alkylation of activated Michael acceptors. The net transformation is the decarboxylative insertion of Michael acceptors into allyl beta-ketoesters. PMID- 15901154 TI - DNA methyltransferase-moderated click chemistry. AB - [reaction: see text]. Biological methylation plays a vital role in regulatory mechanisms of gene transcription. Methylation of both promoter sequences within the genome, as well as protein substrates, has a profound impact upon gene transcription. Yet, few tools exist by which to identify sites of biological methylation in complex biological mixtures. We have generated a novel adenosine derived N-mustard that serves as an efficient synthetic cofactor and allows for subsequent "click" chemistry involving the modified nucleic acid substrate. PMID- 15901155 TI - Lancifodilactone G: a unique nortriterpenoid isolated from Schisandra lancifolia and its anti-HIV activity. AB - [structure: see text]. Lancifodilactone G (1), a novel, highly oxygenated nortriterpenoid featuring a partial enol structure and a spirocyclic moiety, was isolated from the medicinal plant Schisandra lancifolia. Its structure and stereochemistry were determined from extensive one- and two-dimensional NMR and mass spectral data, coupled with single-crystal X-ray analysis. Compound 1 exerted minimal cytotoxicity against C8166 cells (CC50 > 200 microg/mL) and showed anti-HIV activity with EC50 = 95.47 +/- 14.19 microg/mL and a selectivity index in the range of 1.82-2.46. PMID- 15901156 TI - Synthetic studies on the bryostatins: preparation of a truncated BC-ring intermediate by pyran annulation. AB - [reaction: see text]. A synthesis of a potential BC-ring subunit (C9-C27) for bryostatin 1, a remarkably potent anticancer agent, has been developed in 16 steps and 18% overall yield. The key features of this route include a BITIP catalyzed asymmetric allylation reaction, chelation-controlled allylations, a hydroformylation reaction, and a pyran annulation reaction. PMID- 15901157 TI - Synthetic studies on the bryostatins: synthetic routes to analogues containing the tricyclic macrolactone core. AB - [reaction: see text]. Synthesis of the first of a projected series of bryostatin analogues has been accomplished in 26 steps and 2.2% overall yield. In this letter, we detail two approaches to the structural core of these tricyclic macrolactone bryostatin analogues. The key features of the route include BITIP catalyzed asymmetric allylation reactions and Mukaiyama aldol reactions, a chelation-controlled allylation, pyran annulation reactions, and macrolactonization. PMID- 15901158 TI - Radester, a novel inhibitor of the Hsp90 protein folding machinery. AB - [reaction: see text]. The antitumor antibiotics radicicol and geldanamycin are potent inhibitors of the Hsp90 protein folding machinery. Radester is a hybrid composed of radicicol's resorcinol ring and geldanamycin's quinone through an isopropyl ester. Radester was prepared, and the cytotoxicity of it and the corresponding hydroquinone were determined in MCF-7 breast cancer cells to be 13.9 and 7.1 microM, respectively. Protein degradation assays were performed on Hsp90-dependent client proteins, Her-2 and Raf, to correlate Hsp90 inhibition to cytotoxicity. PMID- 15901159 TI - Convenient synthesis of nucleoside and isonucleoside analogues. AB - [reaction: see text]. A very simple methodology to stereoselectively achieve tricyclic isonucleosides (nucleobase = thymine, uracil, and 5-fluoruracil) and 3' C-branched nucleosides (nucleobase = theophylline) was performed by means of a DBU-mediated addition process using a readily available 2-bromo sugar. The mechanism for these transformations implies the loss of both substituents at C-2 and C-3 on the sugar moiety, and although it seems that DBU is probably involved, its involvement has not yet been ascertained. Cytosine did not react under these conditions. PMID- 15901160 TI - Exhaustively methylated azacalix[4]arene: preparation, conformation, and crystal structure with exclusively CH/pi-controlled crystal architecture. AB - [structure: see text]. Described are the preparation, conformation, and crystal structure of exhaustively methylated azacalix[4]arene involving nitrogen atoms as bridging units. NMR and X-ray crystallographic analysis have demonstrated that this novel azacalix[4]arene adopts a 1,3-alternate conformation both in solution and in the solid state. The crystal structure has been characterized solely by intermolecular CH/pi interactions, by which the azacalix[4]arenes mutually interact with each other outside the cavity to furnish a two-dimensional network structure. PMID- 15901161 TI - Heteroannulation of nitroketene N,S-arylaminoacetals with POCl3: a novel highly regioselective synthesis of unsymmetrical 2,3-substituted quinoxalines. AB - [reaction: see text]. A novel regioselective route for the synthesis of substituted and fused 3-chloro-2-(methylthio)quinoxalines through POCl3-mediated heteroannulation of a range of alpha-nitroketene N,S-anilinoacetals has been reported. PMID- 15901162 TI - Regioselective Lewis acid-mediated [1,3] rearrangement of allylvinyl ethers. AB - [reaction: see text]. Aluminum and copper Lewis acids were implemented to effect a regioselective [1,3] rearrangement of allylvinyl ethers in moderate to good yields. The use of trisubstituted alkenes leads to depressed levels of Claisen products. PMID- 15901163 TI - From azides to nitriles. A novel fast transformation made possible by BrF3. AB - [reaction: see text]. Various alkyl and aryl azides, readily obtained from halides or alcohols, were transformed into the corresponding nitriles using bromine trifluoride in moderate to good yields. The reaction is general and gives positive results with aliphatic, aromatic, cyclic, and functionalized azides. It can also be applied to the synthesis of optically active nitriles. PMID- 15901164 TI - Ruthenium-catalyzed intermolecular coupling reactions of arylamines with ethylene and 1,3-dienes: mechanistic insight on hydroamination vs ortho-C-H bond activation. AB - [reaction: see text]. The cationic ruthenium complex [(PCy3)2(CO)(Cl)Ru=CHCH=C(CH3)2]+BF4- was found to be an effective catalyst for the coupling reaction of aniline and ethylene to form a approximately 1:1 ratio of N-ethylaniline and 2-methylquinoline products. The analogous reaction with 1,3 dienes resulted in the preferential formation of Markovnikov addition products. The normal isotope effect of k(NH)/k(ND) = 2.2 (aniline and aniline-d7 at 80 degrees C) and the Hammett rho = -0.43 (correlation of para-substituted p-X C6H4NH2) suggest an N-H bond activation rate-limiting step for the catalytic reaction. PMID- 15901165 TI - Ni(II)-catalyzed Conia-ene reaction of 1,3-dicarbonyl compounds with alkynes. AB - [reaction: see text]. We have discovered a Ni(II)-catalyzed Conia-ene reaction of 1,3-dicarbonyl compounds with alkynes. In the presence of Ni(acac)2 and Yb(OTf)3, various acetylenic 1,3-dicarbonyl compounds underwent Conia-ene reaction to give mono- and bicyclic olefinic cyclopentanes. A mechanism involving the enol-yne-Ni complex formation is proposed and supported by deuterium-labeling experiments. PMID- 15901166 TI - Highly stereocontrolled synthesis of gem-difluoromethylenated azasugars: D- and L 1,4,6-trideoxy-4,4-difluoronojirimycin. AB - [reaction: see text]. D-1,4,6-trideoxy-4,4-difluoronojirimycin and L-1,4,6 trideoxy-4,4-difluoronojirimycin, a novel series of gem-4,4-difluoromethylenated azasugars, were synthesized from CF3CH2OH in 10 steps. A key step was the highly diastereoselective construction of the piperidine ring via reductive amination. PMID- 15901167 TI - N-heterocyclic carbene catalyzed trifluoromethylation of carbonyl compounds. AB - [reaction: see text]. A novel N-heterocyclic carbene (NHC) catalyzed trifluoromethylation reaction of carbonyl compounds was discovered. Both enolizable and nonenolizable aldehydes and alpha-keto esters undergo facile trifluoromethylation with TMSCF3 at room temperature in the presence of only 0.5 1 mol % of the commercially available NHC (1), providing CF3-substituted alcohols in good yields. Selective trifluoromethylation of aldehydes over ketones can be achieved under NHC catalysis. These conditions are mild and simple and tolerate a variety of functional groups. PMID- 15901168 TI - Unique Michael addition-initiated domino reaction for the stereoselective synthesis of functionalized macrolactones from alpha-nitroketones in water. AB - [reaction: see text]. A unique domino reaction of alpha-nitrocycloalkanones with alpha-alkyl alpha,beta-unsaturated aldehydes in aqueous base was discovered, leading to the one-pot synthesis of hitherto unknown functionalized, bridged, bicyclic lactones containing 10-, 11-, 13-, and 15-membered rings. The structures of these heterocyclic compounds, containing also an unusual 6-hydroxy-1,2-oxazine ring, were determined by spectral and single-crystal X-ray diffraction studies. PMID- 15901169 TI - Novel ozone-mediated cleavage of the benzhydryl protecting group from aziridinyl esters. AB - [reaction: see text]. N-Benzhydryl aziridines-2-carboxylates can be readily obtained from the catalytic asymmetric aziridination reaction from N benzhydrylimines and ethyl diazoacetate. Cleavage of the benzhydryl group by hydrogenolysis leads to ring opening when R = aryl. Surprisingly, ozone will selectively oxidize the benhydryl group in these aziridines even when R is an aryl group. This allows for a new deprotection strategy for these aziridines whose generality is explored. PMID- 15901170 TI - Reaction titration: a convenient method for titering reactive hydride agents (Red Al, LiAlH4, DIBALH, L-Selectride, NaH, and KH) by No-D NMR spectroscopy. AB - The concentration of reactive metal hydride (Met-H) reducing agents can be determined (in < or = 20 min) using No-D NMR spectroscopy. The method involves (i) reacting Met-H with an excess of p-methoxybenzaldehyde, (ii) quenching with excess acetic acid, (iii) recording the No-D NMR spectrum of this homogeneous mixture, and (iv) deducing the concentration of Met-H from the % conversion (as measured by integration). By a conceptually related method, the titer of the basic alkali metal hydrides KH and NaH can also be determined. PMID- 15901171 TI - Arylation of diarylamines catalyzed by Ni(II)-PPh3 system. AB - [reaction: see text]. The cross-coupling of bromomagnesium diarylamides, generated in situ from diarylamines, with aryl bromides or iodides can be effected with a simple NiCl2(PPh3)2-PPh3 catalyst system under relatively mild conditions. This coupling reaction is an inexpensive, convenient, and practical method, functioning as an alternative to the corresponding Pd-catalyzed or Cu mediated process for the synthesis of triarylamines. PMID- 15901172 TI - Synthesis of carbazoles from ynamides by intramolecular dehydro Diels-Alder reactions. AB - [reaction: see text]. A new approach to carbazoles and benzannulated carbazoles by means of intramolecular dehydro Diels-Alder reactions of ynamides is reported. By using this approach, carbazoles and benzo[b]-, tetrahydrobenzo[b]-, naphtho[1,2-b]-, naphtho[2,1-b]-, and dibenzo[a,c]carbazoles have been prepared in moderate-to-good yields. PMID- 15901173 TI - Synthesis of nucleoside alpha-thiotriphosphates via an oxathiaphospholane approach. AB - [reaction: see text]. Nucleoside 5'-O-(alpha-thiotriphosphates) were obtained in reactions of the appropriate nucleoside 5'-O-(2-thio-1,3,2-oxathiaphospholanes) with pyrophosphate in the presence of DBU. The presented method allows also for preparation of alpha-seleno congeners and corresponding alpha-modified diphosphates. PMID- 15901174 TI - Enantioselective access to isoquinuclidines by tropenone desymmetrization and homoallylic radical rearrangement: synthesis of (+)-ibogamine. AB - [reaction: see text]. Chiral lithium amide-induced desymmetrization of a tropenone and subsequent Bu3SnH-catalyzed nitrogen-directed homoallylic radical rearrangement constitute key steps in a new strategy to dehydroisoquinuclidines. The strategy was applied in a synthesis of (+)-ibogamine. PMID- 15901175 TI - Synthesis of the C12-C19 fragment of (+)-peloruside A through a diastereomer discriminating RCM reaction. AB - [reaction: see text]. A short and efficient asymmetric synthesis of the C12-C19 fragment of the cytotoxic macrolide (+)-peloruside A has been achieved via a highly diastereomer-discriminating RCM of alpha-branched but-3-enoate ester of a methallylic alcohol derived from hydrolytically resolved (S)-(-)-propylene oxide. PMID- 15901176 TI - Rhodium-catalyzed arylation using arylboron compounds: efficient coupling with aryl halides and unexpected multiple arylation of benzonitrile. AB - [reaction: see text]. The Suzuki-Miyaura-type cross-coupling of arylboron compounds with aryl halides proceeds efficiently in the presence of a rhodium based catalyst system to produce the corresponding biaryls. Furthermore, it has unexpectedly been observed that the treatment with benzonitrile under similar conditions brings about its multiple arylation, in which nucleophilic arylation on the cyano group and subsequent ortho arylation via C-H bond cleavage are involved. PMID- 15901177 TI - Mycapolyols A-F, new cytotoxic metabolites of mixed biogenesis from the marine sponge Mycale izuensis. AB - [structure: see text]. Mycapolyols A-F (1-6), six new unusual PKS/NRPS metabolites, were isolated from the marine sponge Mycale izuensis. The gross structures were elucidated by analysis of spectroscopic data, while the stereochemistry was established using chemical method and the universal NMR database. PMID- 15901178 TI - A sequential palladium-catalyzed alder-ene-reductive amination reaction. AB - [reaction: see text]. Alkyne allyl alcohols are readily transformed into beta amino ethyl alkylidene tetrahydrofurans or beta-amino ethyl alkylidene pyrrolidines in a Pd-catalyzed cycloisomerization-reductive amination sequence with remarkable chemoselectivity leaving benzyl groups and trisubstituted double bonds intact. PMID- 15901179 TI - CO-retentive addition reactions of fluorinated acid chlorides to alkynes. AB - [reaction: see text]. The reaction of perfluorinated acid chlorides with terminal alkynes is efficiently catalyzed by rhodium complexes and proceeds with retention of the CO moiety in the acid chloride to afford (Z)-1-perfluoroacyl-2-chloro-1 alkenes selectively in high yields. PMID- 15901180 TI - Short and efficient route to the fully functionalized polar core of scyphostatin. AB - [reaction: see text]. Diastereoselective oxidative dearomatization of benzopyran 5 to the corresponding p-quinol 9b allows a fast, efficient, and versatile entry to scyphostatin's polar epoxycyclohexenone moiety as demonstrated by the preparation of its palmitoyl analogue 16 (R = (CH2)14CH3). PMID- 15901181 TI - Chelation-controlled intermolecular alkene and alkyne hydroacylation: the utility of beta-thioacetal aldehydes. AB - [reaction: see text]. Beta-thioacetal-substituted aldehydes, which are conveniently prepared from the corresponding ynals, can be combined with a range of alkynes or electron-poor alkenes to deliver intermolecular hydroacylation adducts. The reactions employ [Rh(dppe)]ClO4 as a catalyst and are proposed to proceed via a chelated rhodium acyl intermediate. The thioacetal-containing products can be deprotected to the corresponding ketones or reduced to alkanes in good yields. PMID- 15901182 TI - Efficient dendritic diphosphino Pd(II) catalysts for the Suzuki reaction of chloroarenes. AB - reaction: see text]. The monomeric diphosphino Pd(II) complex 1 and the first three generations of dendritic analogues G1, G2, and G3 are efficient catalysts for the Suzuki coupling reaction of halogenoarenes, including chloroarenes with phenylboronic acid. The recovery and reuse of the dendritic catalysts G1, G2, and G3 are discussed. PMID- 15901183 TI - Unique diketopiperazine dimers from the insect pathogenic fungus Verticillium hemipterigenum BCC 1449. AB - [structure: see text]. Vertihemiptellides A (1) and B (2), unique diketopiperazine dimers, were isolated from the insect pathogenic fungus Verticillium hemipterigenum BCC 1449. Structures of these compounds were elucidated by NMR and mass spectral analysis, and the stereochemistry of 1 was determined by X-ray crystallography. The absolute stereochemistry of bisdethiodi(methylthio)-1-demethylhyalodendrin (3), previously isolated from the same fungus, was revised to the (3R,6R) configuration. PMID- 15901184 TI - Total synthesis of (+)-tubelactomicin A. 1. Stereoselective synthesis of the lower-half segment by an intramolecular Diels-Alder approach. AB - [reaction: see text]. Starting from diethyl (R)-malate, synthesis of the lower half segment of (+)-tubelactomicin A, a 16-membered macrolide antibiotic, has been achieved. The synthesis involved the highly endo- and pi-facial selective intramolecular Diels-Alder reaction achieved using a trisubstituted methacrolein derivative tethering a 10-carbon dienyne unit at the beta-carbon, which in turn was prepared from a known allylated malic acid derivative. PMID- 15901185 TI - Total synthesis of (+)-tubelactomicin A. 2. Synthesis of the upper-half segment and completion of the total synthesis. AB - [reaction: see text]. We have completed the total synthesis of natural (+) tubelactomicin A (1), a 16-membered macrolide antibiotic. This Letter presents a highly efficient synthesis of the upper-half segment (C14-C24) and the completion of the total synthesis featuring a high-yielding Stille coupling for the connection of the upper-half and lower-half segments and Mukaiyama macrolactonization for the construction of the entire structure of 1. PMID- 15901186 TI - Synthesis of fluorous and nonfluorous polycyclic systems by one-pot, double intramolecular 1,3-dipolar cycloaddition of azomethine ylides. AB - [reaction: see text]. Under microwave irradiation, a one-pot, double intramolecular [3 + 2]-cycloaddition reaction of azomethine ylides leads to formation of a novel hexacyclic ring system. The major diastereomer is isolated, and its stereochemistry is determined by X-ray crystal structure analysis. PMID- 15901187 TI - Synthesis of the C11-C23 fragment of spirastrellolide A. A ketal-tethered RCM approach to the construction of spiroketals. AB - [reaction: see text]. The synthesis of the C11-C23 fragment in spirastrellolide A is described here featuring a ketal-tethered RCM as an alternative approach to the construction of spiroketals. PMID- 15901188 TI - 2-O-propargyl ethers: readily cleavable, minimally intrusive protecting groups for beta-mannosyl donors. AB - [reaction: see text]. The use of 2-O-propargyl ethers as protecting groups in 4,6 O-benzylidene-protected mannopyranosyl donors bearing either bulky silyl groups or glycosidic linkages on O3 overcomes the poor stereoselectivity achieved with the corresponding 2-O-benzyl ethers, due to a reduction of steric buttressing. Deprotection is conducted by treatment with potassium tert-butoxide followed by catalytic osmium tetroxide and N-methylmorpholine N-oxide. PMID- 15901189 TI - Sagittamides A and B. Polyacetoxy long-chain acyl amino acids from a didemnid ascidian. AB - [structure: see text]. An unidentified tunicate from Pohnpei, Micronesia, yielded sagittamides A and B-compounds comprising a long-chain C26 dicarboxylic acid that acylates terminal L-valine and L-ornithine groups. The structures, which contain an unprecedented internal O-hexacetyl-1,2,3,4,5,6-hexaol moiety, were determined by combined spectroscopic analysis including mass spectrometry and 1D and 2D NMR and chemical degradation. The partial absolute stereochemistry of the new compounds was addressed by Marfey's analysis. PMID- 15901190 TI - Rhodium-catalyzed asymmetric 1,4-addition of arylboronic acids to coumarins: asymmetric synthesis of (R)-tolterodine. AB - [reaction: see text]. Rhodium-catalyzed asymmetric 1,4-addition of arylboronic acids to coumarins proceeded with high enantioselectivity in the presence of a rhodium catalyst (3 mol %) generated from Rh(acac)(C2H4)2 and (R)-Segphos to give the corresponding (R)-4-arylchroman-2-ones in over 99% ee. This asymmetric reaction was applied to the synthesis of (R)-tolterodine. PMID- 15901191 TI - Tandem Stille/Suzuki-Miyaura coupling of a hetero-bis-metalated diene. Rapid, one pot assembly of polyene systems. AB - [structure: see text]. The synthesis of a hetero-bis-metallo 1,3-butadiene is reported, and its use as an orthogonal Stille and Suzuki-Miyaura coupling partner is detailed. The tin/boron diene participated successfully in a one-pot, sequential Stille and Suzuki-Miyaura coupling protocol, and its utility was demonstrated in the two-step construction of the pentaene side chain of the Fusarium metabolite lucilactaene. PMID- 15901192 TI - The 5th European Conference on Health Economics. PMID- 15901193 TI - Do patients always prefer quicker treatment? : a discrete choice analysis of patients' stated preferences in the London Patient Choice Project. AB - The London Patient Choice Project (LPCP) was established to offer NHS patients more choice over where and when they receive treatment, and to reduce waiting times. The LPCP offered those patients waiting around 6 months for elective procedures a choice of treatment at an alternative NHS or private hospital, or treatment at an overseas hospital.The aim of this article is to investigate the following questions regarding patients' response to choice: (a) What are the factors that patients consider when deciding whether to accept the alternatives they are offered? (b) What is the relative importance to patients of each factor when making their choices, i.e. what trade-offs are patients prepared to make between time waited and other factors? (c) Are there any systematic differences between subgroups of patients (in terms of their personal, health and sociodemographic characteristics) in their response to choice?Patients' preferences were elicited using a discrete choice experiment. Patients eligible to participate in the LPCP were recruited prior to being offered their choice between hospitals and each presented with seven hypothetical choices via a self completed questionnaire. Data were received from 2114 patients. Thirty percent of respondents consistently chose their 'current' over the 'alternative' hospital. All the attributes and levels examined in the experiment were found to exhibit a significant influence on patients' likelihood of opting for an alternative provider, in the expected direction. Age, education and income had an important effect on the 'uptake' of choice. Our results suggest several important implications for policy. First, there may be equity concerns arising from some patient subgroups being more predisposed to accept choice. Second, although reduced waiting time is important to most patients, it is not all that matters. For example, the reputation of the proffered alternatives is of key importance, suggesting careful thought is required about what information on quality and reputation can/should be made available and how it should be made available to facilitate informed choice. PMID- 15901194 TI - Is patient choice an effective mechanism to reduce waiting times? AB - In many countries, patient choice is a routine part of the normal healthcare system. However, many choice initiatives in secondary care are part of policies aimed at reducing waiting times. This article provides evidence on the effectiveness of patient choice as a mechanism to reduce waiting times within a metropolitan area. The London Patient Choice Project was a large-scale pilot offering patients on hospital waiting lists a choice of alternative hospitals with shorter waiting times. A total of 22 500 patients were offered choice and 15 000 accepted. The acceptance rate of 66% was very high by international standards. In this article we address two questions. First, did the introduction of choice significantly reduce waiting times in London relative to the rest of the country where there was no choice? Second, how were the waiting times of London patients not offered choice affected by the choice regime? We examine the evidence on these issues for one specialty, orthopaedics. A difference-in difference analysis is used to compare waiting times for hospitals within London before and after the introduction of choice. Although there was a small but significant reduction in waiting times in London relative to other areas where there was no patient choice, the main effect of the choice regime was to produce convergence of mean waiting times within London. Convergence was achieved by bringing down waiting times at the hospitals with high waiting times to the levels that prevailed in hospitals with low waiting times. This represented a clear improvement in equity of access, an important objective of the English National Health Service. PMID- 15901195 TI - How much might universal health insurance reduce socioeconomic disparities in health? : A comparison of the US and Canada. AB - A strong association between lower socioeconomic status and worse health has been documented within many countries, but little work has been done to compare the strength of this relationship across countries. We compare the strength of the relationship between income and self-reported health in the US and Canada. We find that being below median income raises the likelihood that a middle-aged person is in poor or fair health by about 15 percentage points in the US, compared with less than 8 percentage points in Canada. We also find that this 7 percentage points stronger relationship between low income and poor health in the US compared with Canada is reduced by about 4 percentage points after age 65, the age at which virtually all US citizens receive basic health insurance through the Medicare programme. Income differences in the probability that an individual lacks a usual source of care are also significantly larger in the US than in Canada before the age of 65, but about the same after age 65. Our results are therefore consistent with the theory that the availability of universal health insurance in the US, or at least some other difference that occurs around the age of 65 in one country but not the other, decreases the difference in the strength of the income-health relationship in the US compared with Canada. PMID- 15901196 TI - Senior citizens and the burden of prescription drug outlays: what lessons for the Medicare prescription drug benefit? AB - This article uses data from a cohort of elderly and retired persons over the 1996 2001 period in the US to (i) determine the extent to which changes in socioeconomic or demographic characteristics, particularly age, income and education, impact the total amount that is spent on prescription drugs and (ii) to analyse the predictors of individual out-of-pocket (OOP) prescription drug outlays among the same cohort and determine whether age, race, sex, income, education, marital status and health status have an influence on these. The analysis considers the implications for elderly individuals who choose to participate in the new Medicare Part D drug benefit, labelled the Medicare Prescription Drug, Improvement, and Modernisation Act of 2003. The results highlight the necessity for the Medicare prescription drug benefit to carefully target the eldest among the elderly, who are most in need and are in danger of becoming trapped in the so-called Medicare 'doughnut hole', i.e. incur high prescription drug outlays, without adequate coverage. The study also finds evidence that women, those who are not married, middle income elderly and those in poor health, who purchase drugs more intensively, are at risk of incurring significant prescription OOP drug outlays. PMID- 15901197 TI - Consumer mobility in social health insurance markets : a five-country comparison. AB - During the 1990s, the social health insurance schemes of Germany, the Netherlands, Switzerland, Belgium and Israel were significantly reformed by the introduction of freedom of choice (open enrolment) of health insurer. This was introduced alongside a system of risk adjustment to compensate health insurers for enrolees with predictable high medical expenses. Despite the similarity in the health insurance reforms in these countries, we find that both the rationale behind these reforms and their impact on consumer choice vary widely.In this article we seek to explain the observed variation in switching rates by cross country comparison of the potential determinants of health insurer choice. We conclude that differences in choice setting, and in the net benefits of switching, offer a plausible explanation for the large differences in consumer mobility.Finally, we discuss the policy implications of our cross-country comparison. We argue that the optimal switching rate crucially depends on the goals of the reforms and the quality of the risk-adjustment system. In view of this, we conclude that switching rates are currently too low in the Netherlands, and an active government policy to encourage consumer mobility seems warranted. In Germany and Switzerland, high switching rates call for an improvement of the rather poor risk-adjustment systems. Given low switching rates in Israel and Belgium, improving risk adjustment is less urgent, but still required in the long run. PMID- 15901198 TI - Tackling regional health inequalities in france by resource allocation : a case for complementary instrumental and process-based approaches? AB - This article aims to evaluate the results of two different approaches underlying the attempts to reduce health inequalities in France. In the 'instrumental' approach, resource allocation is based on an indicator to assess the well-being or the quality of life associated with healthcare provision, the argument being that additional resources would respond to needs that could then be treated quickly and efficiently. This governs the distribution of regional hospital budgets. In the second approach, health professionals and users in a given region are involved in a consensus process to define those priorities to be included in programme formulation. This 'procedural' approach is employed in the case of the regional health programmes. In this second approach, the evaluation of the results runs parallel with an analysis of the process using Rawlsian principles, whereas the first approach is based on the classical economic model.At this stage, a pragmatic analysis based on both the comparison of regional hospital budgets during the period 1992-2003 (calculated using a 'RAWP [resource allocation working party]-like' formula) and the evolution of regional health policies through the evaluation of programmes for the prevention of suicide, alcohol-related diseases and cancers provides a partial assessment of the impact of the two types of approaches, the second having a greater effect on the reduction of regional inequalities. PMID- 15901199 TI - Income-related inequality in the use of dental services in Finland. AB - The aim of this article is to measure and explain income-related inequalities in dentist utilisation. We apply concentration and horizontal inequity indices and the decomposition method to decompose observed inequalities into sources. The data are from the Finnish Health Care Survey of 1996. We examine three measures of utilisation: (a) the total number of visits; (b) the probability of visiting a dentist; and (c) the conditional number of positive visits for (i) visits to all dentists, (ii) those to public dentists and (iii) those to private dentists. The results for the whole sample show pro-poor inequities in all three measures of utilisation in public care, whereas in the first two measures there are pro-rich inequities nationwide and in private care. Among those entitled to age-based subsidised dental care, we find equality and equity in all three measures of utilisation nationwide. The two main factors related to pro-rich distributions of use are income and dentist's recall. To enhance equity in dental care across income groups, attention should be focused on supply factors and other incentives to encourage the poor to contact dentists more often. PMID- 15901200 TI - Burning a hole in the budget: tobacco spending and its crowd-out of other goods. AB - Smoking is an expensive habit. Smoking households spend, on average, more than $US1000 annually on cigarettes. When a family member quits, in addition to the former smoker's improved long-term health, families benefit because savings from reduced cigarette expenditures can be allocated to other goods. For households in which some members continue to smoke, smoking expenditures crowd-out other purchases, which may affect other household members, as well as the smoker. We empirically analyse how expenditures on tobacco crowd-out consumption of other goods, estimating the patterns of substitution and complementarity between tobacco products and other categories of household expenditure. We use the Consumer Expenditure Survey data for the years 1995-2001, which we complement with regional price data and state cigarette prices. We estimate a consumer demand system that includes several main expenditure categories (cigarettes, food, alcohol, housing, apparel, transportation, medical care) and controls for socioeconomic variables and other sources of observable heterogeneity. Descriptive data indicate that, comparing smokers to nonsmokers, smokers spend less on housing. Results from the demand system indicate that as the price of cigarettes rises, households increase the quantity of food purchased, and, in some samples, reduce the quantity of apparel and housing purchased. PMID- 15901201 TI - Nurses' labour supply with an endogenous choice of care level and shift type : a nested discrete choice model with nonlinear income. AB - It is argued that increasing wages will not only attract more nurses to the health sector, but also increase the number of hours worked for those already there. This article focuses on the response of registered nurses employed in the public sector when they are allowed to endogenously choose between jobs in hospitals and primary care and between day and shift work. A structural labour supply model is estimated on Norwegian micro-data with job-specific wages and hours. The simulation of an overall public wage increase indicates a reduction in total hours. Thus, in contrast to the claim, the income effect seems to dominate in the labour supply of health sector employees. PMID- 15901202 TI - Testosterone and atherosclerosis in aging men: purported association and clinical implications. AB - Two of the strongest independent risk factors for coronary heart disease (CHD) are increasing age and male sex. Despite a wide variance in CHD mortality between countries, men are consistently twice as likely to die from CHD than their female counterparts. This sex difference has been attributed to a protective effect of female sex hormones, and a deleterious effect of male sex hormones, upon the cardiovascular system. However, little evidence suggests that testosterone exerts cardiovascular harm. In fact, serum levels of testosterone decline with age, and low testosterone is positively associated with other cardiovascular risk factors. Furthermore, testosterone exhibits a number of potential cardioprotective actions. For example, testosterone treatment is reported to reduce serum levels of the pro-inflammatory cytokines interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha, and to increase levels of the anti-inflammatory cytokine IL 10; to reduce vascular cell adhesion molecule (VCAM)-1 expression in aortic endothelial cells; to promote vascular smooth muscle and endothelial cell proliferation; to induce vasodilatation and to improve vascular reactivity, to reduce serum levels of the pro-thrombotic factors plasminogen activator inhibitor (PAI)-1 and fibrinogen; to reduce low-density lipoprotein-cholesterol (LDL-C); to improve insulin sensitivity; and to reduce body mass index and visceral fat mass. These actions of testosterone may confer cardiovascular benefit since testosterone therapy reduces atheroma formation in cholesterol-fed animal models, and reduces myocardial ischemia in men with CHD. Consequently, an alternative hypothesis is that an age-related decline in testosterone contributes to the atherosclerotic process. This is supported by recent findings, which suggest that as many as one in four men with CHD have serum levels of testosterone within the clinically hypogonadal range. Consequently, restoration of serum levels of testosterone via testosterone replacement therapy could offer cardiovascular, as well as other, clinical advantages to these individuals. PMID- 15901203 TI - Anticoagulation during cardioversion in patients with atrial fibrillation: current clinical practice. AB - The role of anticoagulation in the long-term treatment of atrial fibrillation (AF) has been well established in prospective randomized trials. Less certainty exists on the optimal anticoagulation in the setting of AF cardioversion. Current guidelines advocate anticoagulation for 3-4 weeks before and after cardioversion of AF of >48 hours' duration. Alternatively, early cardioversion may be performed after exclusion of left atrial thrombi by transesophageal echocardiography. However, with conventional anticoagulation, the risk of bleeding has to be considered and, thus, anticoagulation is frequently underused in the clinical setting. Moreover, the role of cardioversion has been questioned by recent trials suggesting no benefit of sinus rhythm restoration over rate control in AF. This article aims to summarize the currently available data on anticoagulation in cardioversion of AF in the context of these new studies and points to some new drugs such as low-molecular weight heparins and oral thrombin inhibitors that may lead to safer anticoagulation for prevention of thromboembolic complications of AF in the future. PMID- 15901204 TI - Statins and the response to myocardial injury. AB - The benefits of long-term statin (HMG-CoA reductase inhibitor) treatment for preventing coronary events have been well documented in several large-scale prospective clinical trials. By influencing the determinants of myocardial injury, statins may produce direct cardioprotective effects in the ischemic myocardium and prevent further damaging recurrent events. Although not proven fully in a clinical setting, cholesterol-independent or 'pleiotropic' effects of statins are thought to protect against myocardial injury and may occur via a number of mechanisms. Endothelial dysfunction occurs early in the development of atherosclerosis and is associated with a reduction in endothelial nitric oxide (NO) production. Statins have been shown to increase the expression of endothelial NO synthase, with subsequent augmentation of NO in the vasculature. Statins have also been reported to have anti-inflammatory effects, reducing the release of cytokines and chemokines, decreasing the expression of pro inflammatory cell adhesion molecules, and reducing the accumulation of neutrophils in myocardial tissue following ischemia and reperfusion. Indeed, the role of statins in reducing infarct size is supported by data from a number of preclinical studies. Statin treatment, administered at the onset of reperfusion, has been shown to reduce infarct size by approximately 50% following ischemia in various animal models, and this may be an NO-dependent effect. Randomized clinical trials have indicated that early initiation of statin treatment is associated with a reduction in both the rate of recurrence of cardiovascular events and death in patients with acute coronary syndrome. In addition, decreased rates of myocardial infarction and mortality were demonstrated in several retrospective studies where statin treatment was administered before an interventional procedure. There is a need for further clinical trials to fully elucidate the importance of pre-procedural statin therapy and to determine the extent and mechanisms by which statins exert their cardioprotective effects. PMID- 15901205 TI - Angiotensin II receptor antagonists alone and combined with hydrochlorothiazide: potential benefits beyond the antihypertensive effect. AB - Angiotensin II receptor antagonists (angiotensin receptor blockers; ARBs) and thiazide diuretics have an accepted place in the management of hypertension. Most patients require combination therapy with two or more drugs to adequately control blood pressure to targets recommended by European and international guidelines. ARBs and the thiazide diuretic hydrochlorothiazide have complementary modes of action. Fixed-dose combinations of an ARB and low-dose hydrochlorothiazide provide a convenient and effective treatment option for patients who do not achieve blood pressure targets on monotherapy, without compromising the placebo like tolerability of ARBs. In Europe, fixed-dose combinations with hydrochlorothiazide currently are available for the ARBs candesartan, eprosartan, irbesartan, losartan, telmisartan, and valsartan. Recently, a number of studies have focused on the use of ARBs in monotherapy and in combination therapy, in conditions including congestive heart failure, post-myocardial infarction management, hypertension with cardiovascular risk factors, and diabetic and non diabetic nephropathy. Evidence from these studies suggests a beneficial role beyond the antihypertensive effect of these therapies in providing protection against cardiovascular, renovascular, and cerebrovascular events. PMID- 15901206 TI - Vascular closure devices: a review of their use after invasive procedures. AB - In the endovascular procedure setting, vascular closure devices (VCD) have emerged as an alternative to mechanical compression in order to achieve vascular hemostasis after puncture of the femoral artery. VCD are categorized based primarily on the principle mechanism of hemostasis, which includes biodegradable plug, suture, staples, or ultrasound. While VCD offer advantages over mechanical compression (shorter time to hemostasis and patient ambulation, high rate of patient satisfaction, and greater cost-effectiveness) complications related to the site of femoral access are still present. Efficacy and safety of VCD have been evaluated in a number of clinical trials, but to date there is still a lack of randomized clinical trials with sample sizes large enough to reveal superiority or non-inferiority of VCD compared with mechanical compression. Mechanical compression and VCD are effective and well tolerated in the setting of diagnostic procedures and procedures that do not use anticoagulation. For both methods, success rates are lower, and complication rates higher, in the setting of interventional procedures and when anticoagulation medications are used. Regardless of the device type, deployment failure is the major drawback of VCD. However, overall, meta-analysis data demonstrated that complications and success rates are not significantly different between mechanical compression and VCD. Current data suggest that the correct answer on whether or not to use VCD still remains in the operator domain where integration of clinical data and familiarity with a particular closure device plays a key role in achieving successful hemostasis and avoiding access site complication. PMID- 15901207 TI - Cardiovascular risk factors associated with insulin resistance: effects of oral antidiabetic agents. AB - Patients with type 2 diabetes mellitus have a greater risk of cardiovascular disease than nondiabetic individuals. These patients are often insulin resistant and have an associated clustering of risk factors that contribute to cardiovascular disease. The risk factors include dyslipidemia, hypertension, altered hemostasis, and chronic inflammation. A primary objective in the management of type 2 diabetes mellitus is normalization of blood glucose levels; however, some of the oral drugs used to control blood glucose levels have significant effects on these risk factors. In this article, we review the current data involving the modification of these cardiovascular risk factors by the biguanide (metformin), the thiazolidinediones (troglitazone, rosiglitazone, and pioglitazone), the alpha-glucosidase inhibitors (miglitol, acarbose), and the insulin secretagogs (glyburide [glibenclamide], glipizide, chlorpropamide, tolbutamide, tolazamide, glimepiride, repaglinide, and nateglinide). Generally, the thiazolidinediones improve hemostasis and endothelial function and reduce blood pressure, while having variable effects on dyslipidemia. Metformin improves dyslipidemia and altered hemostasis and decreases plasma C-reactive protein levels with little or no effect on blood pressure. Data on the effects of the alpha-glucosidase inhibitors and insulin secretagogs are sparse; however, these drugs appear to have little or no effect on cardiovascular risk factors. PMID- 15901208 TI - Intravital observation of Plasmodium berghei sporozoite infection of the liver. AB - Plasmodium sporozoite invasion of liver cells has been an extremely elusive event to study. In the prevailing model, sporozoites enter the liver by passing through Kupffer cells, but this model was based solely on incidental observations in fixed specimens and on biochemical and physiological data. To obtain direct information on the dynamics of sporozoite infection of the liver, we infected live mice with red or green fluorescent Plasmodium berghei sporozoites and monitored their behavior using intravital microscopy. Digital recordings show that sporozoites entering a liver lobule abruptly adhere to the sinusoidal cell layer, suggesting a high-affinity interaction. They glide along the sinusoid, with or against the bloodstream, to a Kupffer cell, and, by slowly pushing through a constriction, traverse across the space of Disse. Once inside the liver parenchyma, sporozoites move rapidly for many minutes, traversing several hepatocytes, until ultimately settling within a final one. Migration damage to hepatocytes was confirmed in liver sections, revealing clusters of necrotic hepatocytes adjacent to structurally intact, sporozoite-infected hepatocytes, and by elevated serum alanine aminotransferase activity. In summary, malaria sporozoites bind tightly to the sinusoidal cell layer, cross Kupffer cells, and leave behind a trail of dead hepatocytes when migrating to their final destination in the liver. PMID- 15901209 TI - Interaction of 2-hydroxy-substituted Nile red fluorescent probe with organic nitrogen compounds. AB - The fluorescent properties of 2-hydroxy Nile red dye (HONR) proved to be highly sensitive to the basicity of hydrogen bond acceptors. Fluorescence quantum yields and fluorescence decay profiles were measured as the function of the concentration of organic nitrogen compounds in solvents of various polarities. The detailed mechanism and the kinetics of the fluorescence quenching were revealed with the combined analysis of the steady-state and time-resolved spectroscopic data. The relative contribution of the competing reaction steps was found to be very sensitive to the basicity of the additive and to solvent polarity. The most profound change appeared in the unimolecular deactivation pathways of the excited hydrogen-bonded HONR, whereas the formation rate of this species varied to a lesser extent. The dissociation into excited HONR and ground state base was able to compete with the energy dissipation only when 2,4,6 trimethylpyridine was used as hydrogen bond acceptor in toluene. The bimolecular quenching of the excited hydrogen-bonded complex played significant role in apolar solvents. Proton displacement along the hydrogen bond in the excited complex led to excited ion pairs in polar media. PMID- 15901210 TI - Entropy changes drive the electron transfer reaction of triplet flavin mononucleotide from aromatic amino acids in cation-organized aqueous media. A laser-induced optoacoustic study. AB - The thermodynamic parameters for the formation of the free radicals upon electron transfer quenching of the flavin triplet state (3FMN) by tryptophan and tyrosine, Delta(FR)H and Delta(FR)V, were obtained in aqueous solution by the application of laser-induced optoacoustic spectroscopy at various temperatures. The Delta(FR)H and Delta(FR)V values include the electron transfer and charge separation steps plus the protonation of the FMN anion radical and the deprotonation of the amino-acid cation radical. A linear correlation was found between the Delta(FR)H and Delta(FR)V values for each of the amino acids in phosphate buffers of [CH3(CH2)3]4N+, Li+, NH4+, K+ and Cs+. The compensation between Delta(FR)H and Delta(FR)V within the salt series, and the independent evaluation of the Gibbs energy for electron transfer Delta(ET)G(o) afforded the entropy change, Delta(FR)S, for the reaction, different for the two amino acids. The values of Delta(FR)H, Delta(FR)V and Delta(FR)S in each buffer are mainly determined by the changes in strength and probably number of hydrogen bonds between the reacting partners and water produced along all steps leading to the radicals FMNH* and A*. The Delta(FR)V values linearly correlate with the tabulated entropy of organization of the water structure for the five cations, DeltaS(o)(cat). The entropy change upon formation of the free radicals, Delta(FR)S, quantitatively correlated to the Delta(FR)V value, drives the separation of the ion pair after the electron transfer reaction in the case of highly organizing cations. The ratio X = T Delta(FR)S/Delta(FR)V = (55 +/- 9) kJ cm(-3) for Trp as 3FMN quencher is smaller than X = (83 +/- 9) kJ cm(-3) for Tyr as quencher. These values are discussed in conjunction with the Marcus reorganization energy, as calculated from the Gibbs activation energy of the electron transfer process, which is independent of the salt present but different for each of the two quenchers. PMID- 15901211 TI - Circular dichroism of the photoreceptor pigment oxyblepharismin. AB - Circular dichroism (CD) was used to study the structure of oxyblepharismin (OxyBP), the photoreceptor chromophore for the photophobic response of the blue form of Blepharisma japonicum. Both the chromophore associated to its native protein and the free chromophore in ethanol solution were investigated. CD spectra in the far-UV range indicate that OxyBP induces a slight increase in the alpha-helix content of the protein matrix. CD spectra in the near-UV and visible region of the spectrum show that OxyBP adopts a chiral conformation with a preferential geometry not only when associated to its protein matrix, but also when isolated and dissolved in ethanol. This experimental result is related to the existence of a high-energy interconversion barrier between two enantiomeric structures of the molecule and discussed on the basis of an asymmetric biosynthesis of its precursor, blepharismin. PMID- 15901212 TI - An exploration of the health benefits of factors that help us to thrive. PMID- 15901213 TI - Positive emotions: exploring the other hemisphere in behavioral medicine. AB - The search for the psychological antecedents of medical disorders has focused on the role of stress and negative emotional states. Previous research in this area has investigated relations between negative emotions and physiological adaptations (e.g., blood pressure elevations), adverse health behaviors (e.g., smoking), and social conditions (e.g., social isolation). In this discussion, we argue that more attention is needed to understand the effects of positive emotional states on health enhancement and disease prevention. In each of the areas cited previously, evidence is beginning to emerge that indicates that positive emotions can be associated with health promoting conditions. Interventions using cognitive behavioral strategies or meditation can increase positive emotional states that are maintained over time and that may benefit health and well-being. Implications for behavioral medicine are discussed. PMID- 15901214 TI - The psychobiology of emotion: the role of the oxytocinergic system. AB - A necessary condition for the individual's survival is the capacity for mental, behavioral, and physiological adaptation to external and internal conditions. Consequently, the integrated organism strives to maintain a dynamic, functional balance and integrity under varying conditions. Effective individual adaptation processes are basically dependent on the functioning of the integrated psychophysiological system. In humans, the brain plays a fundamental role in these processes. It serves the adaptation of individuals to current and anticipated conditions by selecting, interpreting, and transforming information into mental, behavioral, and physiological responses. In doing so, the incoming information is linked to existing structures of emotions, values, and goals. Consequently, the interpretation of external information may vary and become subjective depending on an individual's present and past experiences (see e.g., Magnusson, 2003). Hitherto, empirical research has been mainly concerned with the aspect of the psychophysiological system, which is activated in situations that are perceived by the individual as threatening, harmful, or demanding and in which the fight-flight and stress responses described by Cannon (1929) and Selye (1976) play an important role. The aim of this article is to draw attention to a component of the psychophysiological system, the calm and connection system, underlying well-being and socialization. By including this new system, the model of the integrated individual becomes more complete and it enriches the understanding of emotional aspects of brain functioning. PMID- 15901215 TI - Altuism, happiness, and health: it's good to be good. AB - Altruistic (other-regarding) emotions and behaviors are associated with greater well-being, health, and longevity. This article presents a summary and assessment of existing research data on altruism and its relation to mental and physical health. It suggests several complimentary interpretive frameworks, including evolutionary biology, physiological models, and positive psychology. Potential public health implications of this research are discussed, as well as directions for future studies. The article concludes, with some caveats, that a strong correlation exists between the well-being, happiness, health, and longevity of people who are emotionally and behaviorally compassionate, so long as they are not overwhelmed by helping tasks. PMID- 15901216 TI - Life meaning: an important correlate of health in the Hungarian population. AB - One of the 5 coping scales in Rahe's Brief Stress and Coping Inventory, entitled Life Meaning, was examined in relation to demographic characteristics, other coping measures, and health status in a sample of 12,640 Hungarian participants. Participants were selected to represent the country's population according to sex, age, and place of residence. The study also explored the contribution of life meaning to the explanation of variations of middle-aged (45-64 years) male and female mortality rates across 150 subregions in Hungary. On an ecological level life meaning proved to be inversely related to male and female oncological, female cardiovascular, and total premature mortality rates in the 150 subregions of Hungary and on an individual level to participants' reported health status. In the total sample of individuals after controlling for gender, age, and education, life meaning scores showed strong correlations with the World Health Organization well-being scale, with self-rated absence of depression, with self-rated health, and with self-rated absence of disability. Although relatively unrelated to age, gender, and education, life meaning was positively related to self-efficacy, importance of religion, problem-oriented coping, and social support. PMID- 15901217 TI - Dispositional optimism and the mechanisms by which it predicts slower disease progression in HIV: proactive behavior, avoidant coping, and depression. AB - The issue of whether optimism may prospectively protect against disease progression is one that has generated much interest, with mixed results in the literature. The purpose of this study was to determine whether dispositional optimism predicts slower disease progression in HIV. Two indicators of disease progression, CD4 counts and viral load, were assessed over 2 years in a diverse group (men, women, White, African American, Hispanic) of 177 people with HIV in the midrange of disease at entry to the study. Optimism predicted slower disease progression (less decrease in CD4 and less increase in viral load) controlling for baseline CD4 and viral load, antiretroviral treatment, gender, race, education, and drug use. Those low on optimism (25th percentile) lost CD4 cells at a rate 1.55 times faster than those high on optimism (75th percentile). Optimists had higher proactive behavior, less avoidant coping, and less depression: These variables mediated the linear optimism-disease progression relationship. Thus, optimists may reap health benefits partly through behavioral (proactive behavior), cognitive (avoidant coping), and affective (depression) pathways. Implications, limitations, and interpretations are discussed. PMID- 15901218 TI - Sense of coherence and biomarkers of health in 43-year-old women. AB - The aim of this cross-sectional study was to investigate how sense of coherence (SOC) relates to biomarkers of health in 43-year-old nonsmoking premenopausal women. Before taking part in a standardized medical health examination including assessment of blood pressure, blood lipids, and physical symptoms, participants completed a three-item measure of SOC. On the basis of their SOC scores, the 244 women with complete datasets were categorized into 1 of 3 groups with a weak, intermediate, or strong SOC. Results showed that women with a strong SOC had significantly lower levels of systolic blood pressure (p < .05) and total cholesterol (p < .05) than did women with a weak SOC. It is suggested that the lower levels of systolic blood pressure and total cholesterol found in women with a strong SOC may constitute a biological buffer against ill health and disease. PMID- 15901219 TI - Relations between companion animals and self-reported health in older women: cause, effect or artifact? AB - A large longitudinal dataset on women's health in Australia provided the basis of analysis of potential positive health effects of living with a companion animal. Age, living arrangements, and housing all strongly related to both living with companion animals and health. Methodological problems in using data from observational studies to disentangle a potential association in the presence of substantial effects of demographic characteristics are highlighted. Our findings may help to explain some inconsistencies and contradictions in the literature about the health benefits of companion animals, as well as offer suggestions for ways to move forward in future investigations of human-pet relationships. PMID- 15901220 TI - Avoidance and processing as predictors of symptom change and positive growth in an integrative therapy for depression. AB - Depression is a leading cause of disability worldwide and can worsen the course of a variety of medical illnesses. There is a clear need to develop more potent treatments for this debilitating disorder and prevent its return. We are developing a promising psychotherapy that integrates components of current, empirically supported therapies for depression and also teaches healthy lifestyle and emotion regulation habits to promote psychological health. In the 1st open trial, growth curve analyses revealed a significant linear decrease in symptoms of depression in a sample of 29 clients who completed the therapy. Participants wrote essays about their depression each week, and the content was analyzed using a new coding system of change processes. Hierarchical linear modeling (HLM) revealed that peak levels of processing in the essays were associated with more improvement in depression and with the expression of more hope and of both negative and positive views of the self, presumably as clients explored their depressive views of self. Peak levels of avoidance were associated with less improvement in depression and with more hopelessness and negative views of the self. These preliminary results suggest possible targets of change that can facilitate symptom reduction and perhaps also promote psychological health. PMID- 15901222 TI - Temporal concordance of anxiety disorders and child sexual abuse: implications for direct versus artifactual effects of sexual abuse. AB - This study examined the temporal concordance between the onset of childhood anxiety disorders and the points of onset and ending of child sexual abuse (CSA). Sexually abused children (N = 158) were assessed with structured diagnostic interviews. Onset ages for lifetime prevalence anxiety disorders were combined and sequenced with the onset and ending of sexual abuse. Hazard rates were calculated. Departures from the overall linear hazard trajectory for onsets were modeled using piecewise growth curve analyses. Increases from the overall trajectory were found around the point of sexual abuse onset for most childhood anxiety disorders. Decreases were found around the ending of sexual abuse. The risk for developing new anxiety disorders after the onset of sexual abuse showed a positive dose-effect relation with abuse severity. The findings add support to the idea that CSA can have a direct link to childhood anxiety disorders, apart from confounded vulnerability factors, postabuse events, or stable family background factors. The findings are contrasted with those from cross-sectional partial correlation studies that have suggested that there is little direct connection between sexual abuse and mental health outcomes. PMID- 15901223 TI - Children's responses to conflict between their different parents: mothers, stepfathers, nonresident fathers, and nonresident stepmothers. AB - Children who have experienced parental separation have potentially 3 sets of parents whose relationships may impact on them: mother and former partner, mother and stepfather, and father and new partner. Children's accounts of their response to conflict between these different parental dyads were studied, in relation to the quality of their relationships with these parents assessed with child interviews and questionnaires, and to maternal reports of the children's adjustment, in a sample of 159 children growing up in different family settings. Involvement in conflict within 1 parental dyad was chiefly unrelated to such involvement in conflict between the other parental dyads. In contrast, there was evidence for "spillover" effects in relationships within families; for instance, high frequencies of conflict between parents were linked to more troubled parent child relationships. Children were more likely to side with the parent to whom they were biologically related than with stepparents. Involvement in mother nonresident father conflict and in mother-stepfather conflict were both associated with adjustment problems, independent of the qualities of positivity and conflict in the relationship between child and parent. Implications for views on "family boundaries" are considered. PMID- 15901224 TI - The relations among abuse, depression, and adolescents' autobiographical memory. AB - This study examined the relations among early and recent experiences with abuse, depression, and adolescents' autobiographical memory in a longitudinal study of family violence. Participants' (N = 134) exposure to violence was documented when they were 6 to 12 years old and again when they were 12 to 18 years old. The second assessment included measures of depression and autobiographical memory for childhood experiences. Memory problems were more consistently related to current circumstances than childhood abuse history. For instance, depressive symptoms were associated with increased rates of "overgeneral" childhood memories. Recent exposure to family violence predicted more overgeneral memories, shorter memories, and lower rates of negative memories. The patterns suggest that adolescents currently stressed by depression or family violence might strategically avoid the details of past experiences to regulate affect. PMID- 15901225 TI - Childhood traumatic grief: an exploration of the construct in children bereaved on September 11. AB - This study is an exploration of the measurement and correlates of childhood traumatic grief (CTG). Eighty-three children of uniformed service personnel who died during the World Trade Center attack on September 11, 2001, were assessed using measures of demographic characteristics, trauma exposure (physical proximity, emotional proximity, and secondary adversities), use of coping strategies, psychiatric symptoms (posttraumatic stress disorder [PTSD], general anxiety, depression), self-esteem, and traumatic grief. An exploratory factor analysis of the Extended Grief Inventory (EGI; Layne, Savjak, Saltzman, & Pynoos, 2001) indicated distinct constructs of normal versus traumatic grief. CTG factor scores were correlated with secondary adversities from the traumatic event, symptoms of PTSD, anxiety, depression, and coping responses, underscoring the theoretical and clinical utility of the content of the measure. Study limitations and future research recommendations are discussed. PMID- 15901226 TI - Positive parenting as a mediator of the relations between parental psychological distress and mental health problems of parentally bereaved children. AB - This study investigated a positive parenting composite of multiple measures of warmth and consistent discipline as a mediator of the relations between surviving parents' psychological distress and parentally bereaved children's mental health problems using both cross-sectional and prospective longitudinal models. The study included 214 bereaved children ages 7 to 16 and their surviving parent or current caregiver. A multirater, multimethod measurement model of positive parenting was developed. Although the mediational model was supported by analysis of the cross-sectional data, it was not supported in the 3-wave longitudinal model. However, the longitudinal model did find a significant path from positive parenting at Wave 2 to child mental health problems 11 months later at Wave 3, controlling for stability in child mental health problems. Implications for understanding the development of mental health problems of parentally bereaved children are discussed. PMID- 15901227 TI - Positive illusory bias and the self-protective hypothesis in children with learning disabilities. AB - We tested the hypothesis that overestimations of performance by children with learning disabilities (LD) are self-protective and will dissipate following positive feedback. Twenty-three boys and 17 girls with LD (ages 10.6 to 13.5 years) and a control group of non-LD matched children (22 boys and 17 girls) provided a prediction of their performance on a spelling test prior to completing the test. Subsequently, they were randomly assigned to either a positive feedback or a no-feedback condition. Finally, they provided a second prediction of performance on an equivalent spelling test. In children with LD, there was a positive bias in their predictions of performance, and, following positive feedback, their predictions became accurate. In children without LD, there was no positive bias and no effect of feedback. The results provide further support for the presence of a positive illusory bias and for the self-protective hypothesis in children with LD. PMID- 15901228 TI - The measure of Adolescent Heterosocial Competence: development and initial validation. AB - We developed and began construct validation of the Measure of Adolescent Heterosocial Competence (MAHC), a self-report instrument assessing the ability to negotiate effectively a range of challenging other-sex social interactions. Development followed the Goldfried and D'Zurilla (1969) behavioral-analytic model for assessing competence. Approximately 700 adolescents participated in 5 systematic studies. Studies 1 through 4 generated the MAHC item and response content, as well as the basis for response scoring. In the first 3 studies, extensive data were collected from the target population regarding problematic heterosocial situations and potential responses to those situations. In Study 4, expert jdudges rated the relative effectiveness of the adolescent-identified behavioral responses. In the final study, scale reliability and construct validity of the resulting 40-item multiple-choice self-report instrument were examined. Factor analysis yielded no interpretable factors, and the internal consistency of the total scale was acceptable (alpha =.73). Investigation of convergent and discriminant validity revealed that the MAHC was significantly related to measures of general social competence and anxiety in heterosexual situations and was not associated with a measure of socioeconomic status. Contrary to expectations, the MAHC was not correlated with peer ratings of social acceptance. PMID- 15901229 TI - Adolescent motherhood and postpartum depression. AB - Adolescent mothers undergo unique personal and social challenges that may contribute to postpartum functioning. In this exploratory investigation completed within a risk and resilience framework, 149 adolescent mothers, ages 15 to 19, who participated in school-based teen parents' programs, completed measures of parental stress (social isolation and role restriction), maternal competence, weight/shape concerns, and depression. The sample was quite diverse, and no ethnic differences in base rate levels of the variables were detected. Regression analyses indicated that social isolation, maternal competence, and weight/shape concerns predicted unique variance associated with depression level. The findings are discussed in light of future work and the continued need to inform prevention and treatment programs for young mothers. PMID- 15901230 TI - A brief form of the child abuse potential inventory: development and validation. AB - A brief version of the Child Abuse Potential Inventory (CAP) was developed using a development sample of N = 1,470, and cross-validated using an additional sample of N = 713. Items were selected to maximize (a) CAP variance accounted for; (b) prediction of future child protective services reports; (c) item invariance across gender, age, and ethnicity; (d) factor stability; and (e) readability and acceptability. On cross-validation, scores from the resulting 24-item risk scale demonstrated an internal consistency estimate of .89, a stable 7-factor structure, and substantial correlations with the CAP Abuse Risk score (r = .96). The CAP risk cutoff was predicted with 93% sensitivity and 93% specificity (area under the receiver operating characteristics curve = .98), and the Brief Child Abuse Potential Inventory (BCAP) and CAP demonstrated similar patterns of external correlates. The BCAP may be useful as a time-efficient screener for abuse risk. PMID- 15901231 TI - Factor analytic study of the Children's Yale-Brown Obsessive-Compulsive Scale. AB - This study examined the psychometric properties of the Children's Yale-Brown Obsessive-Compulsive Scale (CY-BOCS; Scahill et al., 1997). Participants were 82 children and adolescents diagnosed with obsessive-compulsive disorder (OCD). Confirmatory factor analyses of 2 previously found models (Obsessions and Compulsions; Disturbance and Severity) yielded poor fit indexes. Exploratory factor analysis supported a model of severity and disturbance with slightly different item content than found by McKay et al. (2003). The internal consistency of the factors was acceptable, and the convergent and divergent validity was supported vis-a-vis correlations with clinician ratings of impairment, self-report measures of depression and anxiety, and parent ratings of Tourette's disorder (TD) symptoms. These findings suggest that the CY-BOCS Total Score may represent an inaccurate assessment of symptom severity and supports the use of the revised Severity and Disturbance factors in assessing illness severity. PMID- 15901232 TI - Peer-mediated treatment of socially withdrawn maltreated preschool children: cultivating natural community resources. AB - This study evaluated the effectiveness of Resilient Peer Treatment (RPT). This is a peer-mediated, classroom-based intervention for socially withdrawn, maltreated preschool children. It examined whether the RPT impact generalized from the treatment setting to larger classroom context. Eighty-two maltreated and nonmaltreated, socially withdrawn Head Start children were randomly assigned to either RPT or attention-control (AC) conditions. Data were collected by teachers and independent observers blind to both maltreatment status and treatment condition. Treatment resulted in higher levels of collaborative peer play interactions in the treatment setting posttreatment for both the maltreated and nonmaltreated children. Results documented generalization of the treatment impact to classroom free-play sessions. These findings were supported by teacher ratings of interactive peer play and social skills. PMID- 15901233 TI - The relation between gender role orientation and fear and anxiety in nonclinic referred children. AB - This study examined the relation between gender role orientation and fear and anxiety in a sample of nonclinic-referred children (N = 209) ages 10 to 13 years. Children and their parents completed questionnaires assessing children's gender role orientation, toy and activity preferences, and fear and anxiety. Results generally indicated that femininity and a preference for girls' toys and activities were positively associated with fear and anxiety, whereas masculinity and a preference for boys' toys and activities were negatively related to these emotions. Furthermore, gender role orientation accounted for more of the variance in fear and anxiety scores than the child's sex. PMID- 15901234 TI - Early onset bipolar disorder: clinical and research considerations. AB - This article examined some of the reasons for confusion and controversy surrounding the frequency of diagnosis of bipolar disorder, especially in prepubertal children. Four case vignettes are used to articulate questions surrounding manifestations of euphoria and grandiosity, informant variance, diagnostic implications of medication-induced behavioral toxicity, and treatment implications of family history. Although extant literature cited addresses some of the issues, specific research is needed for definitive answers. PMID- 15901235 TI - Qualitative clinical research with children and adolescents. AB - This article provides an overview of how qualitative research methods (QRMs) can augment the literature in child and adolescent clinical psychology by contributing to theory and hypothesis building. We discuss the utility of qualitative methods in examining the nature of clinical processes and obtaining deeper understandings about quantitative findings. We also present strategies for designing and conducting qualitative investigations, address ethical issues involved in conducting qualitative research with minors, and discuss limitations on inferences that can be made from qualitative findings. PMID- 15901236 TI - Normal-flow virus filtration: detection and assessment of the endpoint in bio processing. AB - The breakthrough of a model virus, bacteriophage PhiX-174, through normal-flow virus filters was studied using both commercial process fluids and model feed streams. The results indicate that (i) PhiX-174 is a reasonable model for a mammalian parvovirus [MMV (murine minute virus)] in virus filtration studies; (ii) PhiX-174 LRV [log(reduction value)] shows a better correlation with percentage flow decline compared with volume processed under a variety of conditions; (iii) although the extent of decline in virus LRV is dependent on the mechanism of filter fouling, the fouling mechanisms operative in a viral validation study are representative of those likely to be found under actual production conditions. The mechanism of LRV decline by many process streams was proposed to be due to selective plugging of small pores. A theoretical model as well as a predictive equation for LRV decline versus flow decay was derived; experimental results from filtration studies using pore-plugging feed stocks were consistent with the equation. As protein solutions may vary in their adsorptive versus plugging behaviour during filtration, an evaluation of the LRV-versus-flow decay relationship on a biopharmaceutical-product-specific basis may be warranted. PMID- 15901238 TI - Characterization of the ATPase activity of topoisomerase II from Leishmania donovani and identification of residues conferring resistance to etoposide. AB - We have cloned and expressed the 43 kDa N-terminal domain of Leishmania donovani topoisomerase II. This protein has an intrinsic ATPase activity and obeys Michaelis-Menten kinetics. Cross-linking studies indicate that the N-terminal domain exists as a dimer both in the presence and absence of nucleotides. Etoposide, an effective antitumour drug, traps eukaryotic DNA topoisomerase II in a covalent complex with DNA. In the present study, we report for the first time that etoposide inhibits the ATPase activity of the recombinant N-terminal domain of L. donovani topoisomerase II. We have modelled the structure of this 43 kDa protein and performed molecular docking analysis with the drug. Mutagenesis of critical amino acids in the vicinity of the ligand-binding pocket reveals less efficient inhibition of the ATPase activity of the enzyme by etoposide. Taken together, these results provide an insight for the development of newer therapeutic agents with specific selectivity. PMID- 15901239 TI - The signal peptide of the rat corticotropin-releasing factor receptor 1 promotes receptor expression but is not essential for establishing a functional receptor. AB - Approximately 5-10% of the GPCRs (G-protein-coupled receptors) contain N-terminal signal peptides that are cleaved off during receptor insertion into the ER (endoplasmic reticulum) membrane by the signal peptidases of the ER. The reason as to why only a subset of GPCRs requires these additional signal peptides is not known. We have recently shown that the signal peptide of the human ET(B)-R (endothelin B receptor) does not influence receptor expression but is necessary for the translocation of the receptor's N-tail across the ER membrane and thus for the establishment of a functional receptor [Kochl, Alken, Rutz, Krause, Oksche, Rosenthal and Schulein (2002) J. Biol. Chem. 277, 16131-16138]. In the present study, we show that the signal peptide of the rat CRF-R1 (corticotropin releasing factor receptor 1) has a different function: a mutant of the CRF-R1 lacking the signal peptide was functional and displayed wild-type properties with respect to ligand binding and activation of adenylate cyclase. However, immunoblot analysis and confocal laser scanning microscopy revealed that the mutant receptor was expressed at 10-fold lower levels than the wild-type receptor. Northern-blot and in vitro transcription translation analyses precluded the possibility that the reduced receptor expression is due to decreased transcription or translation levels. Thus the signal peptide of the CRF-R1 promotes an early step of receptor biogenesis, such as targeting of the nascent chain to the ER membrane and/or the gating of the protein-conducting translocon of the ER membrane. PMID- 15901240 TI - A role for tumour necrosis factor alpha in human small bowel iron transport. AB - Cytokines are integral to the development of anaemia of chronic inflammation. Cytokines modulate hepcidin expression and iron sequestration by the reticuloendothelial system but their direct effects on small bowel iron transport are not well characterized. The aim of the present study was to examine the local effects of TNFalpha (tumour necrosis factor alpha) on small bowel iron transport and on iron transporter expression in the absence of hepcidin. The effects of TNFalpha on iron transport were determined using radiolabelled iron in an established Caco-2 cell model. The effect of TNFalpha on the expression and localization of the enterocyte iron transporters DMT-1 (divalent metal transporter 1), IREG-1 (iron-regulated transporter 1) and ferritin was determined utilizing Caco-2 cells and in a human ex vivo small bowel culture system. TNFalpha mediated an early induction in both iron import and iron export, which were associated with increased DMT-1 and IREG-1 mRNA and protein expression (P<0.05). However, by 24 h, both iron import and iron export were significantly inhibited, coinciding with an induction of ferritin heavy chain (P<0.05) and a decrease in DMT-1 and IREG-1 to baseline levels. In addition, there was a relocalization of IREG-1 away from the basolateral cell border and increased iron deposition in villous enterocytes. In conclusion, TNFalpha has a direct effect on small bowel iron transporter expression and function, leading to an inhibition of iron transport. PMID- 15901241 TI - Effect of ischaemic preconditioning on regional release of inflammatory markers. AB - Systemic markers of inflammation may be increased in patients after percutaneous coronary intervention. In the present study, we evaluated whether IP (ischaemic preconditioning) attenuated inflammation by activating KATP (ATP-sensitive potassium) channels in patients undergoing coronary angioplasty. Patients (n=36) undergoing angioplasty of a major left coronary artery were allocated randomly to one of four groups: a control group, a group receiving nicorandil (an agonist of KATP channels), an IP group or an IP group pretreated with glibenclamide (an antagonist of KATP channels). To measure the release of sCD40L, P-selectin and myeloperoxidase from the ischaemic region, blood samples were drawn simultaneously from the ascending aorta and the great cardiac vein before and 15 min after coronary angioplasty. At 15 min after angioplasty, a significant increase in sCD40L and P-selectin levels in the great cardiac vein in the control group was observed. IP- and nicorandil-treated patients did not show a significant change in sCD40L and P-selectin levels in response to angioplasty. However, the IP-induced attenuation of sCD40L and P-selectin release was abolished by administering glibenclamide. The change in myeloperoxidase levels mirrored those of sCD40L and P-selectin. The levels of inflammatory markers in the aorta remained stable throughout the study. Patients undergoing angioplasty had increased sCD40L and P-selectin levels in the ischaemic region. In conclusion, IP abolished angioplasty-induced myeloperoxidase release by preventing activated platelet-induced P-selectin release via a KATP-channel initiated pathway. Therefore, in addition to its primary effect on cardioprotection, IP may also provide beneficial anti-inflammatory effects on the interaction between platelets and neutrophils. PMID- 15901242 TI - Failure of angiotensin II to suppress plasma renin activity in normotensive subjects with a positive family history of hypertension. AB - The renin-angiotensin system is implicated in the pathophysiology of hypertension. Renin release is regulated by a number of factors, including circulating Ang II (angiotensin II), the so-called short feedback loop. The aim of the present study was to investigate the responsiveness of circulating Ang II on PRA (plasma renin activity) in normotensive subjects with a PFH or NFH (positive or negative family history of hypertension respectively). PRA, renal haemodynamics and urinary sodium excretion were measured during infusion of Ang II without and with pretreatment with the AT1 (Ang II type 1) receptor blocker irbesartan. Normotensive men with a PFH (n=13) and NFH (n=10), with a mean age of 38 years, were given on different occasions intravenous Ang II infusions of 0.1, 0.5 and 1.0 ng.kg-1 of body weight.min-1 before and after pretreatment with 150 mg of irbesartan once a day for 5 consecutive days. RPF (renal plasma flow) and GFR (glomerular filtration rate) were also measured. Before Ang II infusion, the PFH and NFH groups did not differ with respect to BP (blood pressure), body mass index, PRA, RBF (renal blood flow) or urinary sodium. There was no difference in BP or renal haemodynamic response to the highest Ang II dose between the groups. PRA declined with the highest Ang II dose (P<0.01) in subjects with a NFH, but not in subjects with a PFH. After treatment with irbesartan when Ang II had no effect on BP in either group, Ang II also suppressed PRA in subjects with a PFH (P<0.01), and the difference between the groups at baseline was thus eliminated. In conclusion, these findings indicate that subjects with a PFH have a defective Ang II suppression of PRA, which is corrected by AT1 receptor blockade. PMID- 15901243 TI - Aquaporins: highways for cells to recycle water with the outside world. PMID- 15901244 TI - Membrane water transport and aquaporins: looking back. PMID- 15901245 TI - Clinical update on renal aquaporins. AB - Following the discovery of the aquaporin-1 water channel over a decade ago, molecular techniques have been developed to examine the role of renal aquaporin water channels under numerous physiological and pathological conditions. The present article reviews current knowledge regarding the function and dysfunction of renal aquaporins in disorders of water metabolism. PMID- 15901246 TI - Aquaporins from pathogenic protozoan parasites: structure, function and potential for chemotherapy. AB - Infectious diseases, caused by protozoa, such as malaria, sleeping sickness, Chagas' disease or leishmaniasis, are a global threat. The increase in the number of affected individuals and the rapid spread of drug-resistant strains call for specific novel strategies to combat human pathogenic parasites. In the search for novel drug targets, transport proteins for nutrients and metabolites of the parasite-host interface are getting into focus. The present review summarizes and discusses the currently available results on protozoan aquaporins. Various genes coding for aquaporin water and solute channels have been identified in the protozoan genomes and they are probable elements of the parasite's cell membrane. Phylogenetic analysis reveals that individual aquaporin genes are of bacterial or plant origin. So far, six protozoan aquaporins have been cloned and functionally characterized. Typically, these are bifunctional channels and pass water at intermediate to high rates as well as uncharged solutes. In the present review, amino acid compositions of the individual pore entries are compared and permeability properties are attributed to specific protein features. Furthermore, possible physiological roles in osmotic protection and metabolism are discussed. Finally, the potential of protozoan aquaporins for use as a target or entry pathway for chemotherapeutic compounds is reviewed. PMID- 15901247 TI - Linking nuclear mRNP assembly and cytoplasmic destiny. AB - From the very beginning, mRNAs have a complex existence. They are transcribed, capped, spliced, modified at the 3'end, exported from the nucleus, translated, and eventually degraded. These many events not only affect the overall survival and properties of an mRNA, but are also carefully co-ordinated and integrated with quality control mechanisms that function to ensure that only 'proper' mRNAs are translated at the correct developmental time and place. This does not mean that all mRNAs follow a single or uniform path from synthesis to death. Instead, there are diverse means by which the activities of specific mRNAs are regulated, and these controls often depend upon multiple events in the mRNA's life. mRNAs are not found naked in the cell, instead they are part of complex RNPs (ribonucleoproteins) that consist of many factors. These RNPs are highly dynamic structures that change during the lifetime of a given RNA; linking events such as synthesis and processing to the final fate of the mRNA. Here, we will discuss what is known of the assembly of RNPs in general, with specific reference to the myriad of connections between different nuclear events and the cytoplasmic activity of an mRNA. Due to space limitations this review is not comprehensive, instead we focus on specific examples to illustrate these emerging themes in gene expression. PMID- 15901248 TI - Grb10 and Grb14: enigmatic regulators of insulin action--and more? AB - The Grb proteins (growth factor receptor-bound proteins) Grb7, Grb10 and Grb14 constitute a family of structurally related multidomain adapters with diverse cellular functions. Grb10 and Grb14, in particular, have been implicated in the regulation of insulin receptor signalling, whereas Grb7 appears predominantly to be involved in focal adhesion kinase-mediated cell migration. However, at least in vitro, these adapters can bind to a variety of growth factor receptors. The highest identity within the Grb7/10/14 family occurs in the C-terminal SH2 (Src homology 2) domain, which mediates binding to activated receptors. A second well conserved binding domain, BPS [between the PH (pleckstrin homology) and SH2 domains], can act to enhance binding to the IR (insulin receptor). Consistent with a putative adapter function, some non-receptor-binding partners, including protein kinases, have also been identified. Grb10 and Grb14 are widely, but not uniformly, expressed in mammalian tissues, and there are various isoforms of Grb10. Binding of Grb10 or Grb14 to autophosphorylated IR in vitro inhibits tyrosine kinase activity towards other substrates, but studies on cultured cell lines have been conflicting as to whether Grb10 plays a positive or negative role in insulin signalling. Recent gene knockouts in mice have established that Grb10 and Grb14 act as inhibitors of intracellular signalling pathways regulating growth and metabolism, although the phenotypes of the two knockouts are distinct. Ablation of Grb14 enhances insulin action in liver and skeletal muscle and improves whole-body tolerance, with little effect on embryonic growth. Ablation of Grb10 results in disproportionate overgrowth of the embryo and placenta involving unidentified pathways, and also impacts on hepatic glycogen synthesis, and probably on glucose homoeostasis. This review discusses the extent to which previous studies in vitro can account for the observed phenotype of knockout animals, and considers evidence that aberrant function of Grb10 or Grb14 may contribute to disorders of growth and metabolism in humans. PMID- 15901249 TI - Predicting protein subcellular localization: past, present, and future. AB - Functional characterization of every single protein is a major challenge of the post-genomic era. The large-scale analysis of a cell's proteins, proteomics, seeks to provide these proteins with reliable annotations regarding their interaction partners and functions in the cellular machinery. An important step on this way is to determine the subcellular localization of each protein. Eukaryotic cells are divided into subcellular compartments, or organelles. Transport across the membrane into the organelles is a highly regulated and complex cellular process. Predicting the subcellular localization by computational means has been an area of vivid activity during recent years. The publicly available prediction methods differ mainly in four aspects: the underlying biological motivation, the computational method used, localization coverage, and reliability, which are of importance to the user. This review provides a short description of the main events in the protein sorting process and an overview of the most commonly used methods in this field. PMID- 15901250 TI - A brief review of computational gene prediction methods. AB - With the development of genome sequencing for many organisms, more and more raw sequences need to be annotated. Gene prediction by computational methods for finding the location of protein coding regions is one of the essential issues in bioinformatics. Two classes of methods are generally adopted: similarity based searches and ab initio prediction. Here, we review the development of gene prediction methods, summarize the measures for evaluating predictor quality, highlight open problems in this area, and discuss future research directions. PMID- 15901251 TI - Granulometric analysis of spots in DNA microarray images. AB - As the topological properties of each spot in DNA microarray images may vary from one another, we employed granulometries to understand the shape-size content contributed due to a significant intensity value within a spot. Analysis was performed on the microarray image that consisted of 240 spots by using concepts from mathematical morphology. In order to find out indices for each spot and to further classify them, we adopted morphological multiscale openings, which provided microarrays at multiple scales. Successive opened microarrays were subtracted to identify the protrusions that were smaller than the size of structuring element. Spot-wise details, in terms of probability of these observed protrusions, were computed by placing a regularly spaced grid on microarray such that each spot was centered in each grid. Based on the probability of size distribution functions of these protrusions isolated at each level, we estimated the mean size and texture index for each spot. With these characteristics, we classified the spots in a microarray image into bright and dull categories through pattern spectrum and shape-size complexity measures. These segregated spots can be compared with those of hybridization levels. PMID- 15901252 TI - EST-based analysis of gene expression in the porcine brain. AB - Since pig is an important livestock species worldwide, its gene expression has been investigated intensively, but rarely in brain. In order to study gene expression profiles in the pig central nervous system, we sequenced and analyzed 43,122 high-quality 5' end expressed sequence tags (ESTs) from porcine cerebellum, cortex cerebrum, and brain stem cDNA libraries, involving several different prenatal and postnatal developmental stages. The initial ESTs were assembled into 16,101 clusters and compared to protein and nucleic acid databases in GenBank. Of these sequences, 30.6% clusters matched protein databases and represented function known sequences; 75.1% had significant hits to nucleic acid databases and partial represented known function; 73.3% matched known porcine ESTs; and 21.5% had no matches to any known sequences in GenBank. We used the categories defined by the Gene Ontology to survey gene expression in the porcine brain. PMID- 15901253 TI - Fast tree search for a triangular lattice model of protein folding. AB - Using a triangular lattice model to study the designability of protein folding, we overcame the parity problem of previous cubic lattice model and enumerated all the sequences and compact structures on a simple two-dimensional triangular lattice model of size 4+5+6+5+4. We used two types of amino acids, hydrophobic and polar, to make up the sequences, and achieved 2(23)+2(12) different sequences excluding the reverse symmetry sequences. The total string number of distinct compact structures was 219,093, excluding reflection symmetry in the self avoiding path of length 24 triangular lattice model. Based on this model, we applied a fast search algorithm by constructing a cluster tree. The algorithm decreased the computation by computing the objective energy of non-leaf nodes. The parallel experiments proved that the fast tree search algorithm yielded an exponential speed-up in the model of size 4+5+6+5+4. Designability analysis was performed to understand the search result. PMID- 15901254 TI - A novel method for N-terminal acetylation prediction. AB - The NetAcet method has been developed to make predictions of N-terminal acetylation sites, but more information of the data set could be utilized to improve the performance of the model. By employing a new way to extract patterns from sequences and using a sample balancing mechanism, we obtained a correlation coefficient of 0.85, and a sensitivity of 93% on an independent mammalian data set. A web server utilizing this method has been constructed and is available at http://166.111.24.5/acetylation.html. PMID- 15901255 TI - Timeline of genomics (1977-2004). PMID- 15901256 TI - Preterm delivery. PMID- 15901257 TI - Biomarkers for the prediction of preterm delivery. AB - This structured review discusses the current literature on selected biomarkers and their ability to predict preterm delivery (PTD). Among symptomatic women, the likelihood ratio (LR+) for the prediction of PTD was found to be greater than 10 using amniotic fluid (AF) interleukin-6 (IL-6), AF Ureaplasma urealyticum, as well as a multi-marker consisting of cervical IL-6, cervical IL-8, and cervical length (CL). The LR+ was found to be between 5 and 10 for serum C-reactive protein (CRP). An LR+ between 2.5 and 5 was recorded for serum corticotropin releasing hormone (CRH), cervical fetal fibronectin (fFN), cervical IL-6, serum relaxin, and a multi-marker consisting of fFN and CL. CL and bacterial vaginosis (BV) both predicted PTD in women with preterm labor with an LR+ of less than 2.5. In asymptomatic women, AF U. urealyticum and a multimarker consisting of five individual markers [fFN, CL, serum alpha-fetoprotein (AFP), serum alkaline phosphatase, and serum granulocyte colony-stimulating factor (G-CSF)] predicted PTD with an LR+ greater than 10. The LR+ was between 5 and 10 for serum relaxin and CL. LRs+ recorded for serum alkaline phosphatase, salivary estriol, serum CRH, serum G-CSF, cervical IL-6, AF IL-6, cervical fFN, AFP, and Chlamydia all ranged between 2.5 and 5. Finally, an LR+ below 2.5 has been documented for serum ferritin, serum CRP, BV, and cervical ferritin. PMID- 15901258 TI - Progesterone supplementation for preventing preterm birth: a systematic review and meta-analysis. AB - AIM: The aim of this study is to assess the role of progesterone in preterm birth prevention. METHODS: A MEDLINE search (from 1966 to the present; date of last search January 2005) was performed - using the key words progesterone, pregnancy, preterm birth, preterm labor, and randomized, controlled trial - in order to identify randomized, controlled trials in which progesterone (either intramuscular or vaginal administration) was compared with placebo or no treatment. Data were extracted and a meta-analysis was performed. RESULTS: Seven randomized, controlled trials were identified. Women who received progesterone were statistically significantly less likely to give birth before 37 weeks (seven studies, 1020 women, RR = 0.58, 95% CI = 0.48-0.70), to have an infant with birth weight of < or =2.5 kg (six studies, 872 infants, RR = 0.62, 95% CI = 0.49-0.78), or to have an infant diagnosed with intraventricular hemorrhage (one study, 458 infants, RR = 0.25, 95% CI = 0.08-0.82). CONCLUSIONS: For progesterone supplementation to be advocated for women at the risk of preterm birth, the prolongation of gestation demonstrated in this meta-analysis must translate into improved infant outcomes, including a reduction in mortality. There is currently insufficient information to allow recommendations regarding the optimal dose, route, and timing of administration of progesterone supplementation. PMID- 15901259 TI - Prediction of term and preterm parturition and treatment monitoring by measurement of cervical cross-linked collagen using light-induced fluorescence. PMID- 15901260 TI - Vaginal markers of preterm birth. PMID- 15901261 TI - The Danish National Birth Cohort - a data source for studying preterm birth. PMID- 15901262 TI - Recent work on the epidemiology of preterm birth. PMID- 15901263 TI - Cervical length assessment by ultrasound. PMID- 15901264 TI - Serum biomarkers of spontaneous preterm birth. PMID- 15901265 TI - Histologic placental lesions in women with recurrent preterm delivery. AB - BACKGROUND: The aim of this study was to evaluate whether particular placental histopathology lesions are associated with recurrent preterm birth. METHODS: We analyzed a database of 413 consecutive singleton pregnancies delivered at <32 weeks with past reproductive history available. After the exclusion of nulliparous women, the pregnancies were divided according to the obstetrical history into group 1 (n = 328), women without prior preterm delivery (PTD); group 2 (n = 49), women with one prior preterm childbirth; and group 3 (n = 36), women with > or =2 prior preterm deliveries. Demographic and clinical variables were compared among the three groups by using Kruskal-Wallis test and chi-square test for trend. Finally, the individual placental lesions (i.e. 42 lesions of acute or chronic inflammation, uteroplacental vascular pathology, and intraplacental villous lesions) were correlated with the number of prior preterm deliveries by using regression analysis. A two-tailed P < 0.05 was considered significant. RESULTS: No differences were found among the three groups in demographic or clinical variables. Regression analysis of scored placental lesions corrected for gestational age at delivery showed that the number of prior preterm deliveries was correlated only with chronic marginating choriodeciduitis (correlation coefficient = 0.13; P = 0.01) and acute choriodeciduitis (correlation coefficient = 0.14; P = 0.008). CONCLUSIONS: Among women delivered at <32 weeks, those with prior preterm birth have histologic findings compatible with acute or chronic inflammatory involvement of the uterine cavity, suggesting that a prepregnancy endometrial infection rather than an ascending intrapregnancy pathway may be responsible for some recurrences of PTD. PMID- 15901266 TI - Interleukin-6 and interleukin-8 in cervical fluid in a population of Swedish women in preterm labor: relationship to microbial invasion of the amniotic fluid, intra-amniotic inflammation, and preterm delivery. AB - BACKGROUND: Intrauterine infection and inflammation in women with preterm labor are related to adverse perinatal outcome. Due to its subclinical nature, a correct diagnosis depends on retrieval of amniotic fluid. Amniocentesis is, however, not performed as a clinical routine because of its invasiveness. Hypothetically, cytokines in the cervical fluid may represent an alternative diagnostic approach. The aim was to examine cervical interleukin (IL)-6 and IL-8 in relation to microbial invasion of the amniotic fluid, intra-amniotic inflammation, and preterm birth in women in preterm labor. METHODS: Women with singleton pregnancies in preterm labor (<34 weeks of gestation) and intact membranes were included. Cervical (n = 91) and amniotic fluids (n = 56) were collected. Polymerase chain reaction for Ureaplasma urealyticum and Mycoplasma hominis and culture for aerobic and anaerobic bacteria were performed. IL-6 and IL-8 were analyzed with enzyme-linked immunosorbent assay. RESULTS: Non lactobacillus-dominated biota was detected in cervical secretion in 25% (22/89) and the presence of micro-organisms in the amniotic fluid in 16% (9/56) of the patients. The presence of U. urealyticum in the cervical fluid (21/46) was associated with significantly higher levels of IL-6 in the secretion. IL-6 and IL 8 were significantly higher in cervical fluid of women with intra-amniotic infection and inflammation and in women who delivered < or =7 days and/or before 34 weeks of gestation. Cervical IL-6 > or = 1.7 ng/ml was related to intra amniotic inflammation (relative risk: 2.67; range: 1.50-4.74) and had a sensitivity, specificity, positive predictive value, and negative predictive value of 58, 83, 75, and 69%, respectively, in the identification of intra amniotic inflammation. Similar data were obtained for IL-8 > or = 6.7 ng/ml. CONCLUSIONS: High levels of cervical IL-6 and IL-8 are moderately predictive of intrauterine infection/inflammation and preterm delivery. PMID- 15901267 TI - Preterm birth in Sweden 1973-2001: rate, subgroups, and effect of changing patterns in multiple births, maternal age, and smoking. AB - BACKGROUND: The objectives of this report are to evaluate changes in the preterm birth rate in Sweden 1973-2001. Furthermore, describe the proportion of spontaneous and indicated preterm births and assess risk factors for the subgroups of preterm birth during the period from 1991 to 2001. METHODS: A population-based register study of all births occurring in Sweden from 1973 to 2001 registered in the Swedish Medical Birth Register was designed. The analysis of subgroups was restricted to the period 1991-2001. Gestational age was calculated using last menstrual period and best estimate. Odds ratio for preterm birth related to risk factors was calculated for the subgroups' spontaneous and indicated preterm birth. RESULTS: After an increase in the beginning of the 1980s, the preterm birth rate has decreased from 6.3% in 1984 to 5.6% in 2001 (P < 0.0001). The proportion of multiple births born preterm of the total birth rate increased from 0.34% in 1973 to 0.71% in 2001 (P < 0.0001). Spontaneous preterm births account for 55.2% and iatrogenic preterm births for 20.2% of all preterm births. The strongest association with maternal smoking in early pregnancy was found at gestational age <28 weeks and spontaneous preterm birth [odds ratio (OR) smoking versus no smoking: 1.55, 95% confidence intervals (CI): 1.42-1.69]. The strongest association for maternal age was found between gestational age <28 weeks and indicated preterm birth (OR 5-year increase: 1.34, 95% CI: 1.21-1.47). CONCLUSIONS: The preterm birth rate in Sweden has decreased since the mid 1980s. The composition of different subtypes of preterm birth in a Scandinavian low-risk population seems to be similar to populations with higher incidence of preterm birth and perinatal infections. PMID- 15901268 TI - Monocyte chemotactic protein-2 and -3 in amniotic fluid: relationship to microbial invasion of the amniotic cavity, intra-amniotic inflammation and preterm delivery. AB - OBJECTIVE: To evaluate the presence of monocyte chemotactic protein (MCP)-2 and MCP-3 in cervical and amniotic fluid in women in preterm labor. STUDY DESIGN: Cervical and amniotic fluid was sampled from women with singleton pregnancies (< or =34 weeks) in preterm labor (n = 58). RESULTS: Monocyte chemotactic protein-2 (range: 80-583 pg/ml) and MCP-3 (range: 36-649 pg/ml) were detectable in 7/58 women in preterm labor. Monocyte chemotactic protein-3 was found significantly more often in amniotic fluid of women delivered within 7 days (P < 0.001), <34 weeks (P = 0.002), or with intra-amniotic inflammation (P < 0.001) and microbial invasion of the amniotic fluid (P = 0.003). Women in preterm labor had detectable levels of MCP-2 significantly more often if they gave birth before 34 weeks of gestation (P = 0.038) or had intra-amniotic inflammation (P = 0.042). CONCLUSIONS: The presence of MCPs in amniotic fluid of women in preterm labor was associated with preterm birth before 34 weeks of gestation (MCP-2 and MCP-3), microbial invasion (MCP-3), and inflammation (MCP-2 and MCP-3) of the amniotic cavity. PMID- 15901269 TI - Maternal smoking and causes of very preterm birth. AB - BACKGROUND: Smoking increases the risk of preterm birth. The present study was made to elucidate the relation of smoking to causes of very preterm birth. METHODS: In a case-control study on all very preterm births in two regions of Stockholm 1988-1992, prospectively collected data were extracted from antenatal and delivery records on smoking, other maternal characteristics, pregnancy complications, and causes of preterm birth. Cases were live single births with a gestational age of < or =32 weeks and 0 days, and controls were live singletons delivered at 37 weeks or later (n = 295, respectively). Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated with logistic regression. RESULTS: Compared with non-smokers, adjusted ORs of very preterm birth among moderate smokers (1-9 cigarettes per day) and heavy smokers (> or =10 cigarettes per day) were 1.4 (95% CI 0.8-2.4) and 2.9 (95% CI 1.5-5.7), respectively. Compared with non-smokers, risk of preterm labor was increased among moderate and heavy smokers [ORs 1.9 (95% CI 1.0-3.6) and 2.6 (95% CI 1.1-1.6), respectively]. These risks remained essentially unchanged in women without an identifiable cause of preterm labor ('idiopathic preterm labor'). Smoking was also associated with dose dependent increases in risks of preterm birth due to preterm premature rupture of membranes and late pregnancy bleedings. There was no association between smoking and risk of very preterm birth caused by hypertensive diseases. CONCLUSIONS: Smoking increases the risk of very preterm birth caused by preterm labor (including idiopathic preterm labor), preterm premature rupture of membranes, and late pregnancy bleedings. PMID- 15901270 TI - Good perinatal outcome in selective vaginal breech delivery at term. AB - BACKGROUND AND AIM: To compare perinatal outcome in groups of planned vaginal breech delivery, elective cesarean section with the fetus in breech presentation, and planned vaginal delivery with the fetus in cephalic presentation in a university hospital with a tradition of managing breech deliveries by the vaginal route. METHODS: A cohort study from a 7-year period 1995-2002, including 590 planned vaginal deliveries with a term (> 37 weeks) singleton fetus in breech presentation, 396 elective cesarean sections with a term singleton fetus in breech presentation, and 590 control women intending vaginal delivery with a singleton term fetus in cephalic presentation. RESULTS: The Apgar scores were lower in the group of planned vaginal breech delivery, but in other outcome measures there were no significant intergroup differences. The overall neonatal morbidity was small (1.2% vs. 0.5% vs. 0.3% in the respective study groups) if compared to a recently published randomized multicenter study. CONCLUSIONS: Selective vaginal breech deliveries may be safely undertaken in units having a tradition of vaginal breech deliveries. PMID- 15901271 TI - Evidence-based changes in term breech delivery practice in Sweden. AB - OBJECTIVE: Medical documentation on term breech delivery (TBD) advocates planned abdominal delivery based on evidence. The aim of the present study was to describe a change in TBD practice in Sweden following evidence-based documentation arguing in favor of TBD by cesarean section (CS). MATERIALS AND METHODS: The study was a population-based observational study based on data from the Swedish Medical Birth Register. Eligible subjects were all mothers with singleton children in term breech (TB) presentation born between 1999 and 2001 at > 36 weeks' gestational age. Data were processed, and subjects were subdivided into groups, according to mode of delivery. RESULTS: The CS rate increased from 75.3% in 1999 to 86.0% in 2001, due to an increase in planned abdominal deliveries. The change towards abdominal deliveries was more obvious for hospitals with fewer deliveries. While today, an increasing number of hospitals clearly have a non-selective CS policy, with a > 95% CS rate, others still deliver 30% of TB babies vaginally. CONCLUSION: In conclusion, the evidence-based recommendation for TBD has so far had considerable impact on Swedish obstetric practice. PMID- 15901272 TI - Assisted vaginal delivery versus caesarean section in breech presentation. AB - BACKGROUND: The Term Breech Trial (TBT), a well-known study conducted by Hannah and published in the Lancet, revealed a better outcome for neonates after primary caesarean section compared with attempted vaginal delivery. The aim of the present study was to determine whether the results of TBT have to be taken into account when counseling pregnant women in central Europe. METHODS: We investigated 882 women who had delivered infants in breech presentation over a period of 11 years. The neonates had a birthweight of >2500 g and no malformations. We compared mortality and serious neonatal morbidity after attempted vaginal delivery and after primary caesarean section. RESULTS: No infant or maternal mortality was registered in either group. Serious neonatal morbidity was higher (0.5%; n = 2) for attempted vaginal delivery than for primary caesarean section; in the latter group, no child fulfilled the criteria for serious neonatal morbidity. However, the difference was not statistically significant. As expected, after attempted vaginal delivery, the base excess, and 5-min APGAR scores were indicative of more markedly depressed children. CONCLUSION: After careful exclusion of risk factors and informing the patient in detail about the risks and possible complications, vaginal delivery from breech presentation is still warrantable. PMID- 15901273 TI - Term breech delivery in Sweden: mortality relative to fetal presentation and planned mode of delivery. AB - OBJECTIVES: To compare perinatal and infant mortality in breech and cephalic presentations and between planned vaginal and cesarean section (CS) breech deliveries in Sweden. METHODS: The study comprised two parts. Study A is a national cohort study for the period 1991-2001, including 22 549 breech presentations and 875 249 cephalic presentations born at > or =38 completed weeks. Study B is a case-control study, including all 164 breech deliveries with perinatal or 1-year infant death (during 1991-1999 in Sweden) and controls. RESULTS: Study A: Among non-malformed infants, the total mortality rate was 0.46% in breech and 0.28% in cephalic presentations [adjusted odds ratio (OR) 1.6; 95% confidence interval 1.3-1.9]. Non-malformed breech babies were at an increased risk of antenatal death (breech versus cephalic hazard ratio: 2.7, 2.1-3.6). The infant mortality among non-malformed breech deliveries was higher in vaginal birth than in delivery by CS before labor (OR 2.5, 1.2-5.3). The perinatal + infant mortality among non-malformed breech babies was higher at delivery after 39 completed weeks than at CS delivery at 38 weeks (0.53% versus 0.14%; OR 3.5, 1.9-6.4). The estimated needed number of CS to avoid one death was 400. Study B: In breech presentations without malformations, OR for perinatal or infant death was 3.1 (1.7-5.8) at planned vaginal delivery compared with planned CS delivery, and when breech presentations not diagnosed at 37 gestational weeks were excluded, OR was 3.7 (1.6-9.2). CONCLUSIONS: These large population-based and case-control studies both show a significant reduction of perinatal and infant mortality with planned CS in term breech pregnancy. PMID- 15901274 TI - Ovarian actinomycosis in absence of intrauterine contraceptive device: an unusual presentation. PMID- 15901275 TI - Placental chorioangioma with hydrops foetalis: a case report. PMID- 15901276 TI - Bilateral persistent sciatic artery: a potential risk in pelvic arterial embolization for primary postpartum hemorrhage. PMID- 15901277 TI - Uterine artery embolization to control bleeding after myomectomy. PMID- 15901278 TI - Breech deliveries. PMID- 15901280 TI - Molecular, cellular and developmental biology of urothelium as a basis of bladder regeneration. AB - Urinary bladder malfunction and disorders are caused by congenital diseases, trauma, inflammation, radiation, and nerve injuries. Loss of normal bladder function results in urinary tract infection, incontinence, renal failure, and end stage renal dysfunction. In severe cases, bladder augmentation is required using segments of the gastrointestinal tract. However, use of gastrointestinal mucosa can result in complications such as electrolyte imbalance, stone formation, urinary tract infection, mucous production, and malignancy. Recent tissue engineering techniques use acellular grafts, cultured cells combined with biodegradable scaffolds, and cell sheets. These techniques are not all currently applicable for human bladder reconstruction. However, new avenues for bladder reconstruction maybe facilitated by a better understanding of urogenital development, the cellular and molecular biology of urothelium, and cell-cell interactions, which modulate tissue repair, homeostasis, and disease progression. PMID- 15901281 TI - Fibroblast control on epithelial differentiation is gradually lost during in vitro tumor progression. AB - This study aimed to investigate the role of underlying fibroblasts on morphogenesis of in vitro epithelium reconstituted with normal and neoplastic human oral keratinocytes at various stages of malignant transformation. Primary normal human oral keratinocytes (NOKs), early neoplastic/dysplastic human oral keratinocytes (DOK cell line), and neoplastic human oral keratinocytes (PE/CA-PJ 15 cell line) were organotypically grown on top of a collagen type I matrix with or without primary normal human oral fibroblasts. Morphogenesis of the reconstituted epithelia was assessed by histomorphometry, immunohistochemistry (Ki-67, cyclin D1, cytokeratin 13 (CK13), collagen IV, E-cadherin, p53, CD40), and the terminal deoxynucleotidyl transferase-mediated dUTP in situ nick end labelling method. Reproducible in vitro models of multistage oral carcinogenesis were established. Presence of fibroblasts in the collagen matrix significantly increased cell proliferation in all three models (p<0.05), and induced an invasive pattern of growth in the neoplastic cell lines (p<0.05). In normal, but not in neoplastic oral keratinocytes fibroblasts induced the expression of CD40, and polarized the expression of E-cadherin and p53 to the basal cell layer. In both normal and early neoplastic keratinocytes (DOK cell line), fibroblasts induced the expression of CK13 and collagen IV. In the neoplastic oral keratinocytes (PE/CA-PJ 15 cell line), the presence of underlying fibroblasts did not change the expression of any of the protein markers assessed. This study showed that (1) major steps of oral carcinogenesis can be reproduced in vitro, and (2) the tight control exerted by fibroblasts on epithelial morphogenesis of in vitro reconstituted normal human oral mucosa is gradually lost during neoplastic progression. PMID- 15901282 TI - Frizzled-7 receptor ectodomain expression in a colon cancer cell line induces morphological change and attenuates tumor growth. AB - Frizzled (FZD) receptors have a conserved N-terminal extracellular cysteine-rich domain that interacts with Wnts and co-expression of the receptor ectodomain can antagonize FZD-mediated signalling. Using the ectodomain as an antagonist we have modulated endogenous FZD7 signalling in the moderately differentiated colon adenocarcinoma cell line, SK-CO-1. Unlike the parental cell line, which grows as tightly associated adherent cell clusters, the FZD7 ectodomain expressing cells display a spread out morphology and grow as a monolayer in tissue culture. This transition in morphology was associated with decreased levels of plasma membrane associated E-cadherin and beta-catenin, localized increased levels of vimentin and redistribution of alpha6 integrin to cellular processes in the FZD7 ectodomain expressing cells. The morphological and phenotype changes induced by FZD7 ectodomain expression in SK-CO-1 cells is thus consistent with the cells undergoing an epithelial-to-mesenchymal-like transition. Furthermore, initiation of tumor formation in a xenograft tumor growth assay was attenuated in the FZD7 ectodomain expressing cells. Our results indicate a pivotal role for endogenous FZD7 in morphology transitions that are associated with colon tumor initiation and progression. PMID- 15901283 TI - PTHrP promotes murine secondary trophoblast giant cell differentiation through induction of endocycle, upregulation of giant-cell-promoting transcription factors and suppression of other trophoblast cell types. AB - The murine trophoblast cell lineage represents an intriguing experimental cell model as it is composed of four trophoblast stem (TS)-derived cell types: trophoblast giant cells (TGCs), spongiotrophoblast, syncytotrophoblast, and glycogen trophoblast cells. To investigate the role of parathyroid hormone related protein (PTHrP) in TGC differentiation, we analyzed the effect of exogenous PTHrP on secondary TGCs of day 8.5 p.c. ectoplacental cone explant culture. Secondary TGCs expressed PTHrP and PTHR1 receptor in vivo and in vitro. TGCs treated with PTHrP had reduced proliferation and decreased apoptosis starting from day 2 in culture, and enhanced properties of giant cell differentiation: increased DNA synthesis, number of cells with giant nuclei and expression of placental lactogen-II (PL-II). The induction of TGC formation by PTHrP correlated with downregulation of cyclin B1 and mSNA expression, but upregulation of cyclin D1, thus allowing mitotic-endocycle transition. Moreover, PTHrP treatment influenced TGC differentiation by inducing the expression of transcription factors known to stimulate giant cell formation: Stra13 and AP 2gamma, and inhibiting the formation of other trophoblast cell types by suppressing trophoblast progenitors and spongiotrophoblast-promoting factors, Eomes, Mash-2, and mSNA. Taken together with the spatial and temporal patterns of TGC formation and PTHrP synthesis in vivo, these findings indicate an important role for PTHrP in the differentiation of secondary TGCs during placentation. PMID- 15901284 TI - Cytoskeletal dynamics of the teleostean fin ray during fin epimorphic regeneration. AB - Teleost fishes can regenerate their fins by epimorphic regeneration, a process that involves the transition of the formerly quiescent tissues of the stump to an active, growing state. This involves dynamic modifications of cell phenotype and behavior that must rely on alterations of the cytoskeleton. We have studied the spatial and temporal distribution of three main components of the cytoskeleton (actin, keratin and vimentin) in the regenerating fin, in order to establish putative relationships between cell cytoskeleton and cell behavior. According to our results, the massive rearrangement undergone by the epidermis right after injury, which takes place by cell migration, correlates with a transient down regulation of keratin and a strong up-regulation of actin in the epidermal cells. During the subsequent epidermal growth, based on cell proliferation, keratin normal pattern is recovered while actin is down-regulated, although not to normal (quiescent) levels. The epidermal basal layer in contact with the blastema displays a particular cytoskeletal profile, different to that of the rest of the epidermal cells, which reflects its special features. In the connective tissue compartment, somatic cells do not contain vimentin, but keratin, as intermediate filament. Proliferative and migrative activation of these cells after injury correlates with actin up-regulation. Although this initial activation does not involve keratin down-regulation, blastemal cells were later observed to lack keratin, suggesting that such cytoskeletal modification might be needed for connective tissue cells to dedifferentiate and form the blastema. Cell differentiation in the newly formed, regenerated ray is accompanied by actin down regulation and keratin up-regulation. PMID- 15901285 TI - Male sterility in mice lacking retinoic acid receptor alpha involves specific abnormalities in spermiogenesis. AB - The severe degeneration of the germinal epithelium and subsequent male sterility observed in mice null for the retinoic acid receptor alpha (RARalpha) gene suggested its critical role in spermatogenesis, although the etiology and progression of these abnormalities remain to be determined. Previous studies have revealed that elongated spermatids in RARalpha(-/-) testes were improperly aligned at the tubular lumen and did not undergo spermiation at stage VIII(*). We now report a distinctive failure of step 8-9 spermatids to orient properly with regard to the basal aspect of Sertoli cells, resulting in stage VIII(*)-IX(*) tubules with randomly oriented spermatids. By in situ terminal deoxynucleotidyltransferase-mediated deoxy-UTP nick end labeling (TUNEL), we noted that elongating spermatids frequently underwent apoptosis. Immunohistochemical analysis revealed that while activated caspase-3, the primary effector caspase in the apoptotic cell death machinery, was detected in the nuclei of primary spermatocytes in the first wave of spermatogenesis and occasionally in spermatogonia of both normal and mutant testes, it was not involved in the death of elongating spermatids in RARalpha(-/-) testes. Thus, sterility in RARalpha(-/-) males was associated with specific defects in spermiogenesis, which may correlate with a failure in both spermatid release and spermatid orientation to the basal aspect of Sertoli cells at stage VIII(*) in young adult RARalpha(-/-) testis. Further, the resulting apoptosis in elongating spermatids appears to involve pathways other than that mediated by activated caspase-3. PMID- 15901286 TI - Catheterization-Doppler discrepancies in nonsimultaneous evaluations of aortic stenosis. AB - Prior validation studies have established that simultaneously measured catheter (cath) and Doppler mean pressure gradients (MPG) correlate closely in evaluation of aortic stenosis (AS). In clinical practice, however, cath and Doppler are rarely performed simultaneously; which may lead to discrepant results. Accordingly, our aim was to ascertain agreement between these methods and investigate factors associated with discrepant results. We reviewed findings in 100 consecutive evaluations for AS performed in 97 patients (mean age 72 +/- 10 yr) in which cath and Doppler were performed within 6 weeks. We recorded MPG, aortic valve area (AVA), cardiac output, and ejection fraction (EF) by both methods. Aortic root diameter, left ventricular end-diastolic dimension (LVIDd) and posterior wall thickness (PWT) were measured by echocardiography and gender, heart rate, and heart rhythm were also recorded. An MPG discrepancy was defined as an intrapatient difference > 10 mmHg. Mean pressure gradients by cath and Doppler were 36 +/- 22 mmHg and 37 +/- 20 mmHg, respectively (P = 0.73). Linear regression showed good correlation (r = 0.82) between the techniques. An MPG discrepancy was found in 36 (36%) of 100 evaluations; in 19 (53%) of 36 evaluations MPG by Doppler was higher than cath, and in 17 (47%) of 36, it was lower. In 33 evaluations, EF differed by >10% between techniques. Linear regression analyses revealed that EF difference between studies was a significant predictor of MPG discrepancy (P = 0.004). Women had significantly higher MPG than men by both cath and Doppler (43 +/- 25 mmHg versus 29 +/- 15 mmHg [P = 0.001]; 42 +/- 23 mmHg versus 32 +/- 15 mmHg [P = 0.014], respectively). Women exhibited discrepant results in 23 (47%) of 49 evaluations versus 13 (25%) of 51 evaluations in men (P = 0.037). After adjustment for women's higher MPG, there was no statistically significant difference in MPG discrepancy between genders (P = 0.22). No significant interactions between MPG and aortic root diameter, relative wall thickness (RWT), heart rate, heart rhythm, cardiac output, and time interval between studies were found. In clinical practice, significant discrepancies in MPG were common when cath and Doppler are performed nonsimultaneously. No systematic bias was observed and Doppler results were as likely yield lower as higher MPGs than cath. EF difference was a significant predictor of discrepant MPG. Aortic root diameter, relative wall thickness, heart rate, heart rhythm, cardiac output, presence or severity of coronary artery disease, and time interval between studies were not predictors of discrepant results. PMID- 15901287 TI - Correlation between right ventricular indices and clinical improvement in epoprostenol treated pulmonary hypertension patients. AB - The aim of this study was to evaluate which parameter of right ventricular (RV) echocardiographic best mirrors the clinical status of patients with pulmonary arterial hypertension. Patients with pulmonary arterial hypertension on epoprostenol therapy were identified via hospital registry. Twenty patients, (16 females, 4 males) were included in the study, 9 with primary pulmonary hypertension and 11 with other diseases. Echocardiograms before therapy and at 22.7 (+/-9.3) months into therapy were compared. The right ventricular myocardial performance index (RVMPI) was measured as the sum of the isometric contraction time and the isometric relaxation time divided by right ventricular ejection time. Other measures included peak tricuspid regurgitation jet velocity (TRV), pulmonary artery systolic pressure (PASP), pulmonary valve velocity time integral (PVVTI), PASP/PVVTI (as an index of total pulmonary resistance) and symptoms by New York Heart Association (NYHA) functional class. Echo parameters of right ventricular function were analyzed in patients, before and during therapy. There was significant improvement of NYHA class in patients following epoprostenol therapy (P < 0.0001). Peak tricuspid regurgitant jet velocity (pre 4.2 +/- 0.6 m/sec, post 3.8 +/- 0.7 m/sec, P = 0.02) and PASP/PVVTI (pre 6.7 +/- 3.3 mmHg/m per second, post 4.8 +/- 2.2 mmHg/m per second, P < 0.0001) were significantly improved during treatment. RVMPI did not improve (pre 0.6 +/- 0.3, post 0.6 +/- 0.3, P = 0.54). Changes in NYHA class did not correlate with changes in RVMPI (P = 0.33) or changes in PASP/PVVTI (P = 0.58). Despite significant improvements in TRV, PASP/PVVTI, and NYHA class, there was no significant change in RVMPI on epoprostenol therapy. Changes in right ventricular indices were not correlated with changes in NYHA class. PMID- 15901288 TI - Color M-mode flow propagation velocity and conventional doppler indices in the assessment of diastolic left ventricular function during isometric exercise. AB - BACKGROUND AND OBJECTIVE: Color M-mode flow propagation velocity (Vp) was shown to be a preload-independent measure of diastolic function. To study the effects of an increase in afterload induced by isometric handgrip exercise on diastolic function assessment in patients with cardiomyopathy, we measured Vp and conventional Doppler indices at baseline and at 30% of predetermined maximum handgrip strength. METHODS: Twenty-four patients with systolic dysfunction were divided into two groups: Group I comprising 12 patients with E/A < 1 (early filling velocity/atrial contraction velocity) and Group II comprising 12 patients with E/A > 1. All the patients underwent measurement of Vp, E velocity, its deceleration time (DT), A velocity, isovolumic relaxation time (IVRT), and pulmonary atrial flow reversal velocity (PFR) at baseline and at 30% of predetermined maximum handgrip strength. Twelve healthy controls underwent these same measurements. RESULTS: When comparing baseline to peak echocardiographic data, no significant changes were noted in Vp in any of the groups while a shift of pulsed Doppler indices of Group I toward a pattern closer to that of Group II was noted and a decrease in E velocity and E/A ratio with an increase in IVRT occurred in healthy controls. CONCLUSIONS: Color M-mode flow propagation velocity seems to be an afterload-independent measure of diastolic function in patients with moderate to severe cardiomyopathy while pulsed Doppler indices are more sensitive to loading conditions induced by isometric exercise. PMID- 15901289 TI - Contrast echocardiography can save nondiagnostic exams in mechanically ventilated patients. AB - Patients in an intensive care unit (ICU) under mechanical ventilation (MV) are very difficult to image by transthoracic echocardiography, diminishing the beneficial information that could be obtained by this noninvasive approach. The objective of this study is to assess whether the addition of a contrast agent to fundamental imaging (FI) can improve or change the initial diagnosis in cardiac postoperative patients under mechanical ventilation by enhancing endocardial border delineation and Doppler flow signal. Thirty mechanically ventilated post cardiac surgery patients (20 men, mean age 61 +/- 13 years) were evaluated with FI before and after intravenous injection of contrast. Left ventricular endocardial border delineation score index (EBDSI), estimated left ventricular ejection fraction (LVEF), and color and spectral Doppler were analyzed. The use of contrast resulted in a significant increase in the number of well-delineated segments, with a salvage rate of 77% of nondiagnostic studies. EBDSI was 1.62 +/- 0.61, before contrast, increasing to 2.05 +/- 0.53 after it (P < 0.001). There was a change in the LVEF estimation in 5 exams, and a new wall motion abnormality was detected in other 4 exams, after the use of contrast. Moreover, a significant change was observed in the quantification of mitral regurgitation in 5 patients, in the aortic transvalvular peak gradient in 1 patient, and measurement of tricuspid regurgitation peak flow velocity in 8 patients. It is concluded that in cardiac postoperative patients under mechanical ventilation, intravenous injection of a contrast agent using FI resulted in a high salvage rate of studies and changed the initial diagnosis in a significant number of patients. PMID- 15901290 TI - Pulmonary transit of echocontrast agents during mechanical ventilation: a clinical transthoracic echocardiographic study. AB - In this study, we investigated whether the ultrasound contrast agents Levovist or Sono Vue injected intravenously during mechanical ventilation effectively pass through the pulmonary circulation. With echocardiography, we measured the time for the contrast to pass through the lungs; and the intensity of right and left ventricular cavity opacification at four time points: during spontaneous breathing (baseline), 5 minutes after the beginning of mechanical ventilation, and 5 minutes and 30 minutes after extubation. Forty patients undergoing elective peripheral neurosurgical procedures were prospectively and randomly enrolled: 20 patients received intravenous Levovist 1 g and 20 patients received intravenous Sono Vue 1 mL, at the four predefined time points. After intravenous injection, both Levovist and Sono Vue effectively passed through the lungs and opacified the right and left ventricular cavities, at the four time points. Pulmonary transit times were similar and constant for the two contrast agents tested: 6 +/- 2 seconds at baseline, 5 +/- 2 seconds during mechanical ventilation, 7 +/- 2 seconds at 5 minutes and 6 +/- 2 seconds at 30 minutes after extubation with Levovist; and 6 +/- 4 seconds at baseline, 6 +/- 3 seconds during mechanical ventilation, 6 +/- 2 seconds at 5 minutes and 7 +/- 3 seconds at 30 minutes after extubation with Sono Vue. In all patients, each of the four contrast injections achieved high-grade right and left ventricular chamber opacification. In conclusion, both the ultrasound contrast agents tested in this study, Levovist and Sono Vue, after intravenous injection pass through the pulmonary circulation during mechanical ventilation. Ultrasound contrast agents with these characteristics are suitable for intraoperative organ perfusion studies, with intravenous injection. PMID- 15901291 TI - Stunning of the left atrium after conversion of atrial fibrillation: predictor for maintenance of sinus rhythm? AB - Recurrence of atrial fibrillation (AF) after cardioversion (CV) to sinus rhythm (SR) is determined by various clinical and echocardiographic parameters. The aim of this study was to determine the value of mitral inflow A-wave velocity, performed at 24 hours after CV in estimation of AF recurrence. The study group consisted of 187 consecutive patients with nonvalvular atrial fibrillation, who had been cardioverted to SR from 1998 to 2000. Transthoracic echocardiography was performed in all cases recruited for the study 24 hours after CV. Left atrial (LA) diameter, left ventricular ejection fraction, and mitral inflow A-wave velocity were measured. The patients were evaluated in five groups according to their recurrence time (<30 days, 31-90 days, 91-180 days, 181-365 days, and >365 days). Maintenance of SR was determined to have a negative linear correlation with age (r =-0.97, P = 0.006), LA diameter (r =-0.93, P = 0.02), and AF duration (r =-0.93, P = 0.02), while having a positive linear correlation with mitral inflow A-wave velocity (r = 0.96, P = 0.008). In the maintenance of sinus rhythm, age, LA diameter, and AF duration were not affected from the method of CV, while mitral inflow A-wave velocity was found to be affected with the method of CV. No relationship was determined between mitral inflow A-wave velocity and the maintenance of sinus rhythm in those performed electrical cardioversion, while frequency of recurrence was found to be higher in those with slow mitral inflow A wave velocity who were performed pharmacological cardioversion (r = 0.89, P = 0.004). In conclusion, age, duration of AF, LA diameter, and the mitral inflow A wave velocity can be used to predict the maintenance of SR after CV. PMID- 15901292 TI - The importance of increased interatrial septal thickness in patients with atrial fibrillation: a transesophageal echocardiographic study. AB - BACKGROUND: Histological studies in animal models have showed that extensive atrial fibrosis or fatty deposition as a result of loss of atrial cardiomyocytes increases the propensity to develop atrial fibrillation (AF). Although several reports have suggested that AF in humans may be a consequence of these mechanisms, no study has correlated the presence of AF with interatrial septal thickness (IAST). METHODS: Accordingly, we conducted a prospective analysis in 150 consecutive patients referred for transesophageal echocardiography (TEE) for numerous medical reasons. A total of 105 patients (mean age 62 +/- 14 years) who met inclusion and exclusion criteria were included for analysis. Pertinent demographics, echocardiographic chamber dimensions, left ventricular ejection fraction (LVEF), color flow, and spectral Doppler analysis, as well as IAST measured in both systole (S) and diastole (D) were obtained. RESULTS: When patients with a documented history of AF were compared to patients without documented AF, no statistical difference was noted in terms of patients' height, weight, IVS thickness, or LVEF. Patients with AF had a significantly larger left and right atrial dimensions (P < 0.001), lower left atrial appendage emptying velocities (P < 0.002), and pulmonary vein systolic Doppler signal (P < 0.01). The IAST in systole in patients with AF was 0.75 +/- 0.27 cm versus 0.60 +/- 0.16 cm in patients without AF (P < 0.006) while the IAST in diastole was 0.61 +/- 0.22 cm versus 0.49 +/- 0.12 cm, respectively (P < 0.009). However, no statistical difference was noted between IAST in either systole (P < 0.8) or diastole (P < 0.8) among patients with AF based on the duration of this arrhythmia. CONCLUSIONS: The results of this prospective TEE study show a statistically significant increase in IAST with the presence of AF independent of patient's age, height, weight, and the degree of IVS thickness. In addition, since no significant valvular abnormalities or compromise in left ventricular systolic function were present, the increase in IAST in patients with AF then suggest possible changes in the material properties of the atrial wall, easily identified by TEE on the interatrial septum, either as a cause or as a result of AF. Since no correlation was found between the degree of IAST and the duration of AF, the presence of IAST not only might identify patients with a higher propensity to have or develop this atrial arrhythmia; but also be a surrogate marker of changes within the components of the atrial wall in AF. PMID- 15901293 TI - Immediate and long-term effects of color Doppler ultrasound on myocardial cell apoptosis of fetal rats. AB - The objective of this article is to evaluate the safety of diagnostic color Doppler ultrasound (CDUS) on embryos by observing its effects on myocardial cell apoptosis of fetal rats. Pregnant Sprague-Dawley rats were divided into fetal group and neonatal group according to the sample-drawing time averagely and randomly, and each group was subdivided into control group and insonification group. The control group was sham-insonificated, and the insonification group was insonificated by diagnostic ultrasound (3.0 MHz, Tis = 1.8, MI = 1.6) for 30 minutes. The fetal rats' hearts were removed 24 hours after insonification and the neonatal rats' hearts were removed 10 days after birth. Apoptosis cells were detected with TUNEL (in situ terminal deoxynucleotidyl transferase-mediated D-UTP nick end labeling), and ultrastructural changes were observed with transmission electron microscope. Myocardial cell apoptosis was significantly higher in the fetal insonification group than in the fetal control group (P < 0.05), and it was significantly higher in the fetal groups than in the neonatal groups (P < 0.05), but there was no significant difference in myocardial cell apoptosis between the neonatal groups (P < 0.05). Apoptotic changes of myocardial cells in the fetal insonification group were observed with transmission electron microscope, which showed margination, condensation of the nuclear chromatin, etc. It is concluded that apoptosis and ultrastructural changes could be induced if fetal rat's heart was irradiated continuously over 30 minutes by diagnostic CDUS, but this phenomenon would disappear after birth. PMID- 15901294 TI - A rare case of Lactobacillus acidophilus presenting as mitral valve bacterial endocarditis. AB - Lactobacillus acidophilus is a gram-positive rod that is a commensal of human mucosal tissues. They are usually considered nonpathogenic flora of the mouth, gut, and female genital tract. Lactobacillus is a rare cause of endocarditis with less than 50 cases reported in the world literature to date. We report the case of a 63-year-old woman who developed native mitral valve endocarditis secondary to L. acidophilus, and examine the literature regarding this rare entity. PMID- 15901295 TI - Atrial endocarditis--the importance of the regurgitant jet lesion. AB - The jet lesions of valvular regurgitation or intracardiac shunts have been hypothesized to play an important role in the pathogenesis of endocarditis for many years. We describe a case of mitral valve endocarditis that involved the left atrium along the path of a jet lesion. This resulted in atrial endocarditis and pericarditis, both of which complicated her presentation and hospital course. Using transesophageal echocardiography, we were able to directly visualize the path and full extent of infection prior to surgery. Special attention should be focused upon the path of eccentric jets in order to fully define the extent of endocarditis. PMID- 15901296 TI - Continuous intraoperative transesophageal echocardiography during pericardiectomy for constrictive pericarditis revealing dynamic change in chamber size. AB - We review a case of a 50-year-old man with diagnosis of constrictive pericarditis, who underwent pericardial stripping with continuous intraoperative transesophageal echocardiography (TEE). This patient demonstrates the immediate dynamic changes in chamber size after successful surgical removal of pericardium as demonstrated by TEE. PMID- 15901297 TI - Mitral annular subvalvular left ventricular aneurysm. AB - Annular subvalvular left ventricular aneurysm was first reported in 1962. This type of aneurysm usually arises from the annular subaortic or submitral region of the left ventricle. It should be differentiated from the left ventricular false aneurysm, which was caused by myocardial necrosis. The etiology of subvalvular aneurysm remains unclear. We have presented a case of annular submitral left ventricular aneurysm. The patient had no history of coronary artery disease. Two dimensional echocardiography and magnetic resonance image (MRI) showed a huge left ventricular aneurysm existed. An annular submitral left ventricular aneurysm was confirmed by an open heart surgery and pathological examination. PMID- 15901298 TI - Free-floating left atrial ball thrombus following mitral valve replacement. AB - We report the case of a 51-year-old woman who underwent mitral valve replacement for prolapse with severe regurgitation, depressed ejection fraction, and atrial fibrillation. Two weeks post-operatively, a transesophageal echocardiogram was performed for bacteremia. The patient was found incidentally to have a large free floating ball thrombus in the left atrium. The patient was managed with anticoagulation because of the high-risk nature of repeat surgery. One month following diagnosis, the patient still had persistent thrombus in the left atrium seen on transthoracic echocardiography despite therapeutic anticoagulation. Free floating ball thrombus is a rare and dramatic finding seen on echocardiography in patients with mitral valve disease. PMID- 15901299 TI - A right atrial mass and a pseudomass. AB - Right atrial (RA) masses are rare entities often detected incidentally during imaging studies. Leading etiologies of right atrial masses are tumor, thrombi, and vegetations. We present two cases of right atrial masses, a cardiac lipoma and an artifact. Clinical and echocardiographic characteristics of benign cardiac tumors are reviewed. We then highlight the importance of considering artifact in the differential diagnosis of atrial masses. Finally, we discuss echocardiographic characteristics of right atrial masses that may provide clues for diagnosis. Right atrial masses, often detected incidentally during imaging studies, are uncommon and can be due to many etiologies including tumors, thrombus, vegetations, normal variants, and artifacts. We describe 2 patients with RA masses detected on routine transthoracic echocardiogram. PMID- 15901300 TI - Live three-dimensional transthoracic echocardiographic assessment of thrombus in the innominate veins and superior vena cava utilizing right parasternal and supraclavicular approaches. AB - We report the usefulness of right parasternal and supraclavicular live three dimensional transthoracic echocardiography in the delineation and follow-up of a thrombus involving a catheter placed in superior vena cava for dialysis in an adult patient with chronic renal disease. PMID- 15901301 TI - Multiple causes of left ventricular outflow tract obstruction in a 58-year-old woman. PMID- 15901302 TI - Compression of intrapericardial structures in tamponade, including "new" echocardiographic manifestations. PMID- 15901303 TI - Commonly used respiratory and pharmacologic interventions in the echocardiography laboratory. PMID- 15901304 TI - Papillary fibroelastoma and Lambl's excrescences: echocardiographic diagnosis and differential diagnosis. PMID- 15901305 TI - Profiling of liquid crystal displays with Raman spectroscopy: preprocessing of spectra. AB - Raman spectroscopy is applied for characterizing paintable displays. Few other options than Raman spectroscopy exist for doing so because of the liquid nature of functional materials. The challenge is to develop a method that can be used for estimating the composition of a single display cell on the basis of the collected three-dimensional Raman spectra. A classical least squares (CLS) model is used to model the measured spectra. It is shown that spectral preprocessing is a necessary and critical step for obtaining a good CLS model and reliable compositional profiles. Different kinds of preprocessing are explained. For each data set the type and amount of preprocessing may be different. This is shown using two data sets measured on essentially the same type of display cell, but under different experimental conditions. For model validation three criteria are introduced: mean sum of squares of residuals, percentage of unexplained information (PUN), and average residual curve. It is shown that the decision about the best combination of preprocessing techniques cannot be based only on overall error indicators (such as PUN). In addition, local residual analysis must be done and the feasibility of the extracted profiles should be taken into account. PMID- 15901306 TI - Phase transition in poly(vinylidene fluoride) investigated with micro-Raman spectroscopy. AB - The phase transition from the non-polar alpha-phase to the polar beta-phase of poly(vinylidene fluoride) (PVDF) has been investigated using micro-Raman spectroscopy, which is advantageous because it is a nondestructive technique. Films of alpha-PVDF were subjected to stretching under controlled rates at 80 degrees C, while the transition to beta-PVDF was monitored by the decrease in the Raman band at 794 cm(-1) characteristic of the alpha-phase, along with the concomitant increase in the 839 cm(-1) band characteristic of the beta-phase. The alpha-->beta transition in our PVDF samples could be achieved even for the sample stretched to twice (2x-stretched) the initial length and it did not depend on the stretching rate in the range between 2.0 and 7.0 mm/min. These conclusions were corroborated by differential scanning calorimetry (DSC) and X-ray diffraction experiments for PVDF samples processed under the same conditions as in the Raman scattering measurements. Poling with negative corona discharge was found to affect the alpha-PVDF morphology, improving the Raman bands related to this crystalline phase. This effect is minimized for films stretched to higher ratios. Significantly, corona-induced effects could not be observed with the other experimental techniques, i.e., X-ray diffraction and infrared spectroscopy. PMID- 15901307 TI - Method for quantitative determination of spatial polymer distribution in alginate beads using Raman spectroscopy. AB - A new method based on Raman spectroscopy is presented for non-invasive, quantitative determination of the spatial polymer distribution in alginate beads of approximately 4 mm diameter. With the experimental setup, a two-dimensional image is created along a thin measuring line through the bead comprising one spatial and one spectral dimension. For quantitative analysis of the Raman spectra, the method of indirect hard modeling was applied to make use of the information contained in the entire recorded spectra. For quantification of the alginate signals from within the beads, a calibration curve acquired from sodium alginate solutions was used after it was shown that only negligible differences occur between signals from alginate solutions and alginate gels. The distribution of alginate over the bead gel matrix was acquired with high spatial (51 microm) and time (12 s) resolution. The inhomogeneous distribution obtained using the new measuring technique is qualitatively in excellent agreement with data from the literature. In contrast to known measuring techniques, correct quantitative information about the spatial polymer distribution within the matrix was derived. It gave an alginate mass fraction of approximately 0.045 g/g at the edges and 0.02 g/g in the center of the beads. Next to the determination of mere polymer concentrations, the excellent time resolution of the presented method will enable investigation of the dynamic process of gel formation and it will also serve as a basis for investigation of mass transfer of small diffusing molecules in alginate matrices. PMID- 15901308 TI - Red-excitation dispersive Raman spectroscopy is a suitable technique for solid state analysis of respirable pharmaceutical powders. AB - Dispersive Raman spectroscopy with excitation by a red diode laser is suitable for quantitative crystallinity measurements in powders for pulmonary drug delivery. In spray-dried mixtures of salmon calcitonin and mannitol, all three crystalline polymorphs of mannitol and amorphous mannitol were unambiguously identified and their mass fractions were measured with a limit of quantification of about 5%. The instrument design offered high sensitivity and adequate background suppression, resulting in a low limit of detection in the range of 0.01% to 1%. This spectroscopy method has significant advantages over established techniques regarding specificity, sensitivity, and sample requirements. PMID- 15901309 TI - Noninvasive method for the assessment of dermal uptake of pesticides using attenuated total reflectance infrared spectroscopy. AB - Dermal absorption of pesticides is a primary exposure route for agricultural workers, but is not well characterized. Current measurement techniques are either invasive, such as tape-stripping, or require extensive sample preparation or analysis time, such as urinary metabolite monitoring or wipe sampling followed by gas chromatography analysis. We present the application of a noninvasive, spectroscopic approach for the measurement of pesticide absorption into skin. Attenuated total reflectance infrared spectroscopy (ATR-IR) was used to monitor directly the absorption of two pesticides--captan and azinphos-methyl--in ten volunteers over 20 min under occlusive conditions. We found substantial variability in absorption across subjects. Our results were comparable to those measured by the more traditional method of wipe-sampling followed by extraction and gas chromatography analysis. Multivariate data analysis, in the form of multivariate curve resolution (MCR), is a novel addition to this type of experiment, yielding time-resolved information unachievable by standard methods. These data are potentially more informative than the monitoring of blood or urinary metabolites because they can be acquired in essentially real-time, allowing observations of pesticide absorption on a rapid timescale rather than over hours or days. PMID- 15901310 TI - Determination of the percentage of homopolymer component in Ziegler/Natta catalyst linear low-density polyethylene resins using high-temperature cell Fourier transform infrared and partial least squares quantitative analysis technique. AB - A new method for the determination of the percentage of homopolymer component, using high-temperature cell Fourier transform infrared (FT-IR) by partial least squares (PLS) quantitative analysis technique, was developed and applied to Ziegler Natta linear low-density polyethylene (LLDPE). The method is based on the IR spectrum changes between the 730 cm(-1) band and 720 cm(-1) band at the temperature of 110 degrees C, which is near the melting point of the polyethylene. The HD % (the percentage of high-density component, i.e., the percentage of homopolymer component) results obtained by CTREF (CRYSTAF in TREF mode) technique are used as the input data together with the respective FT-IR spectra for PLS analyses to establish a calibration curve. The PLS quality is characterized by a correlation coefficient of 0.997 (cross-validation) using four factors and a root mean square error of calibration (RMSEC) of 0.772. The HD% of the unknown can then be predicted by the PLS software from the unknown FT-IR spectrum. A control resin was tested seven times by CTREF and FT-IR. The HD% of the control resin was 28.59+/-0.88% by CTREF and 29.05+/-2.37% by FT-IR. It was found that the method was applicable for the same comonomer type of LLDPE within a melt index range and density. PMID- 15901311 TI - Dynamic infrared linear dichroism study of the temperature-dependent, viscoelastic behavior of a poly(ester urethane). AB - In the study reported here, of the poly(ester urethane), Estane 5703, simultaneous dynamic mechanical analysis (DMA) and dynamic infrared linear dichroism (DIRLD) measurements have been carried out at continuously variable temperatures from -50 to +30 degrees C. Multivariate curve resolution-alternating least squares (MCR-ALS) analysis of the spectral data has been correlated with the thermo-mechanical properties. Spectral changes, analyzed as a function of temperature, are compared with the storage and loss moduli to provide insight into viscoelastic behavior at the molecular level. In addition, the data for pure Estane have been compared to those for plasticized Estane samples, which contain 10 and 30% plasticizer by weight. These comparisons show a strong and consistent correlation between the macroscopic rheological properties and the microscopic (molecular, inter-molecular, and sub-molecular) responses of this block co polymer. PMID- 15901312 TI - Phase correction of interferograms using digital all-pass filters. AB - A method for the phase correction of interferograms in Fourier transform infrared spectroscopy is presented. It is shown that phase error can be canceled to within an arbitrary angular precision by a low-order digital all-pass filter. Such a filter only modifies the phase of the Fourier transform of the interferogram and keeps the magnitude unchanged, like the Mertz method, for example. However, our method minimizes the asymmetric apodization that results in photometric errors when using the Mertz method alone. A practical example is provided in which phase correction over a frequency range of 800 cm(-1) to 4000 cm(-1) using a 9-pole all pass filter resulted in a photometric error of <0.01%, much less than the 0.3% error of the Mertz method. An alternative and faster (approximately 100 ms) approach is to use an all-pass filter with lower angular precision followed by the Mertz method. Removing most of the phase error with the filter brings the interferogram to an optimal state so that the residual phase error can be completely removed with the Mertz procedure without introducing photometric error. The method can be used in most experiments, including emission spectroscopy, where conventional techniques are inadequate. A simple all-pass filter design algorithm is given. PMID- 15901313 TI - Orientation distribution functions based upon both (P1), (P3) order parameters and upon the four (P1) up to (P4) values: application to an electrically poled nonlinear optical azopolymer film. AB - The most probable orientational distribution functions of rod-like polar molecules contained in a noncentrosymmetric uniaxial system are established using the first-rank and third-rank Legendre polynomials, (P1(cos theta)) and (P3(cos theta)) order parameters, and the maximum entropy method. Emphasis is put on the different domains of existence in the ((P1), (P3)) plane for the various shapes of the distributions: it is thus shown that, for any positive (P1(cos theta)) value and for decreasing (P3(cos theta)) values, the distribution function may exhibit either a distorted oblate form with an intense maximum at 0 degrees, or a three-leaved rose curve with maxima at 60 degrees, 180 degrees, and 300 degrees, and finally another markedly oblate shape with a strong maximum at 180 degrees. As an illustrative example, we have considered the azobenzene molecular orientations in an electrically poled p(DR1M) homopolymer thin film after a thermal process and several relaxation periods. We have made use not only of the (P1) and (P3) parameters determined from polarized second-harmonic generation (SHG) measurements, but also of the (P2) values extracted from UV-visible spectra and of the (P4) values adjusted according to the information entropy theory. In such a thin film with very large nonlinear properties (d33 coefficients were varying from 437.0 to 117.0 pm/V at 1064 nm) it is evidenced that a strong polar order is maintained even after a long relaxation period of 42 days. So, the distribution functions demonstrate that the poling treatment was quite efficient and they emphasize the importance in the determination of both couples of odd and even order parameters in such uniaxially oriented optical elements. PMID- 15901314 TI - Equilibrium modeling of mixtures of methanol and water. AB - An understanding of the species that form in mixtures of alcohol and water is important for their use in liquid chromatography applications. In reverse-phase liquid chromatography the retention of solutes on a chromatography column is influenced by the composition of the mobile phase, and in the case of alcohol and water mobile phases, the amount of free alcohol and water present. Previous and similar modeling studies of methanol (MeOH) and water mixtures by near-infrared (NIR) spectroscopy have found up to four species present including free MeOH and water and MeOH and water complexes formed by hydrogen bonding associations. In this work an equilibrium model has been applied to NIR measurements of MeOH and water mixtures. A high-performance liquid chromatography (HPLC) pump was coupled to an NIR flow cell to produce a gradual change in mixture composition. This resulted in a greater mixture resolution than has been achieved previously by manual mixture preparation. It was determined that five species contributed to the data. An equilibria model consisting of MeOH, H2O, MeOH(H2O) (log K(MeOH)H2O = 0.10+/-0.03), MeOH(H2O)4 (log K(MeOH)4H2O = -2.14+/-0.08), and MeOH(H2O)9 (log K(MeOH)9H2O = -8.6+/-0.1) was successfully fitted to the data. The model supports the results of previous work and highlights the progressive formation of MeOH and water complexes that occur with changing mixture composition. The model also supports that mixtures of MeOH and water are not simple binary mixtures and that this is responsible for observed deviations from expected elution behavior. PMID- 15901315 TI - Multiplex single strand conformation polymorphism analysis by capillary electrophoresis with on-the-fly fluorescence lifetime detection. AB - This paper describes the use of on-the-fly fluorescence lifetime detection (OFLD) for multiplex single strand conformation polymorphism (SSCP) analysis by capillary electrophoresis (CE). The dye labels studied for multiplex SSCP-OFLD-CE analyses included RG, NBD, and BODIPY-FL. The dyes were first investigated for a model system of "Wild Type" and "Mutant" 43-base fragments designed to vary by a single A/T substitution. Two dye pairs, BODIPY-FL/ RG and BODIPY-FL/NBD, were then used to detect the G20210A mutation in the human prothrombin gene. Mobility correction was required for the BODIPY-FL/RG system. Three "blind" analyses were performed of three mixtures that combined a control fragment (wild type-BODIPY FL) with two "unknown" fragments selected among four possibilities (wild type or mutant labeled with NBD or RG). In each multiplex analysis, the "origin" of the unknown fragments was correctly identified on the basis of fluorescence lifetime of the dye label and the presence or absence of the mutation was correctly determined on the basis of conformation-induced differences in migration time. PMID- 15901316 TI - Comparison of near-infrared and laser-induced breakdown spectroscopy for determination of magnesium stearate in pharmaceutical powders and solid dosage forms. AB - Near-infrared (NIR) spectroscopy has become well established in both the pharmaceutical arena and other areas as a useful technique for rapid quantitative analysis of solid materials. Though laser-induced breakdown spectroscopy (LIBS) has not been widely applied in the pharmaceutical industry, the technique has been used for rapid quantitative analysis of solids in many other applications. One analysis amenable to each technique is the determination of magnesium stearate in solids during the lubrication blending unit operation of pharmaceutical processing. A comparative study of the utility of these two techniques for this application will be presented. Necessary sample preparations and the extent and type of matrix effects will be discussed. Additionally, it will be shown that NIR provides better accuracy and precision than LIBS with the experimental parameters used; however, LIBS showed superior selectivity as it was demonstrated to be more robust to sample matrix perturbations. Examples of blending applications will also be presented. PMID- 15901317 TI - Hydrogen leak detection using laser-induced breakdown spectroscopy. AB - Laser-induced breakdown spectroscopy (LIBS) is investigated as a technique for real-time monitoring of hydrogen gas. Two methodologies were examined: The use of a 100 mJ laser pulse to create a laser-induced breakdown directly in a sample gas stream, and the use of a 55 mJ laser pulse to create a laser-induced plasma on a solid substrate surface, with the expanding plasma sampling the gas stream. Various metals were analyzed as candidate substrate surfaces, including aluminum, copper, molybdenum, stainless steel, titanium, and tungsten. Stainless steel was selected, and a detailed analysis of hydrogen detection in binary mixtures of nitrogen and hydrogen at atmospheric pressure was performed. Both the gaseous plasma and the plasma initiated on the stainless steel surface generated comparable hydrogen emission signals, using the 656.28 Halpha emission line, and exhibited excellent signal linearity. The limit of detection is about 20 ppm (mass) as determined for both methodologies, with the solid-initiated plasma yielding a slightly better value. Overall, LIBS is concluded to be a viable candidate for hydrogen sensing, offering a combination of high sensitivity with a technique that is well suited to implementation in field environments. PMID- 15901318 TI - 1H and 13C nuclear magnetic resonance studies of the hindered phencyclone adducts of some smaller branched N-alkyl maleimides: rigorous aryl proton assignments with high-resolution two-dimensional (COSY45) spectroscopy, and anisotropic shielding effects and ab initio geometry optimizations. AB - Phencyclone, 1, a potent Diels-Alder diene, reacts with a series of N alkylmaleimides, 2, to form hindered adducts, 3. The 300 MHz 1H and 75 MHz 13C NMR studies of these adducts at ambient temperatures have demonstrated slow rotations on the nuclear magnetic resonance (NMR) timescales for the unsubstituted bridgehead phenyl groups, and have revealed substantial magnetic anisotropic shielding effects in the 1H spectra of the N-alkyl groups of the adducts. The selected N-alkyl groups for the target compounds emphasized smaller branched alkyls, including C3 (isopropyl, a); C4 (isobutyl, b; and t-butyl, c); C5 (n-pentyl, d; isopentyl [isoamyl], e; 1-ethylpropyl, f; t-amyl, g;) and a related C8 isomer (1,1,3,3-tetramethylbutyl ["t-octyl"], h). The straight-chain n pentyl analog was included as a reference. This present work on the branched N alkylmaleimide adducts appreciably extends our earlier compilation on the N-n alkylmaleimide adducts. Key methods for proton assignments included "high resolution" 1H-1H chemical shift correlation spectroscopy, COSY45. 13C NMR of the adducts, 3, verified the expected number of aryl carbons for slow exchange limit (SEL) spectra of the bridgehead phenyl groups. The synthetic routes involved reaction of the corresponding amines, 4, with maleic anhydride to give the N alkylmaleamic acids, 5, which underwent cyclodehydration to form the maleimides, 2. Magnetic anisotropic shielding magnitudes for alkyl group protons in the adducts were calculated relative to corresponding proton chemical shifts in the maleimides. Geometry optimizations for the above adducts (and for the N-n butylmaleimide adduct) were performed at the Hartree-Fock level with the 6-31G* basis set. The existence of different contributing conformers for the adducts is discussed with respect to their calculated energies and implications regarding experimentally observed anisotropic shielding magnitudes. PMID- 15901319 TI - Diels-alder adducts of 3,6-dibromophencyclone with symmetrical 1,4-disubstituted cis-2-butenes: comparisons with the adduct of phencyclone and N-benzylmaleimide, and one-dimensional and two-dimensional nuclear magnetic resonance studies and ab initio structure calculations. AB - Hindered Diels-Alder adducts have been prepared from 3,6-dibromophencyclone, 2, with cis-1,4-diacetoxy-2-butene, 3; cis-2-butene-1,4-diol, 4; and N benzylmaleimide, 5. The adduct from the parent phencyclone, 1, with N benzylmaleimide was prepared for comparison. One- and two-dimensional (1D and 2D) proton and carbon-13 NMR studies (at 7.05 tesla, ambient temperatures), including high-resolution COSY45 and HETCOR (XHCORR) chemical shift correlation spectra, were performed, allowing extensive rigorous assignments for protons and protonated carbons. Substantial anisotropic shielding was seen for the ortho protons of the N-benzyl group in the adducts of 5 with 1 or 2, with these aryl protons resonating at 6.25 ppm (CDCl3) for each adduct. The unsubstituted bridgehead phenyls of all four adducts showed slow exchange limit (SEL) 1H and 13C spectra. Greater shift dispersions for the bridge-head phenyl protons in the adducts from 5 relative to those from 3 or 4 suggested the role of the imide carbonyls for anisotropic contributions or for influences on adduct geometry. Ab initio geometry optimizations were performed at the Hartree-Fock level with the 6 31G* basis set (or the LACVP* basis set for the bromine-containing compounds) for each of the adducts. For the two adducts from benzylmaleimide, separate minima were located corresponding to conformers in which the benzyl group was directed into the adduct cavity (syn) or out of the adduct cavity (anti). Calculated energies and geometric parameters for the adducts are presented, and these suggested a significantly different structure for the dibromo diacetate adduct, in terms of general symmetry and bridgehead phenyl geometries, compared to the other adducts. PMID- 15901320 TI - Diels-alder adducts of 3,6-dibromophencyclone with short-chain N-n alkylmaleimides: 1H and 13C nuclear magnetic resonance studies of hindered rotations and magnetic anisotropy, and ab initio calculations for optimized structures. AB - 3,6-Dibromophencyclone, 2, reacted with N-ethylmaleimide, 3a; N-n propylmaleimide, 3b; and N-n-butylmaleimide, 3c; to form the corresponding Diels Alder adducts, 4a, 4b, and 4c. The nuclear magnetic resonance (NMR) spectra of the adducts were studied at ambient temperatures at 300 MHz for proton and 75 MHz for carbon-13. Full proton assignments were achieved by high-resolution COSY45 spectra for the aryl proton regions. Rigorous assignments for protonated carbons were obtained with the heteronuclear chemical shift correlation spectra (HETCOR). Slow exchange limit (SEL) spectra were observed for both proton and carbon-13 NMR for each adduct, with slow rotation on the NMR timescales for the unsubstituted bridgehead phenyl groups. Endo Diels-Alder adduct stereochemistry was supported by substantial magnetic anisotropic shielding effects in the 1H NMR spectra of the alkyl groups. Proton NMR shifts are compared with those previously reported for the corresponding adducts, 5b and 5c, obtained from 3b and 3c, respectively, with the parent compound, phencyclone, 1. Results of ab initio molecular modeling calculations at the Hartree-Fock level using the LACVP* basis set for conformers of the dibrominated adducts, 4a-4c, are presented, together with HF/6-31G* results for the non-brominated adducts, 5a, 5b, and 5c. Novel aspects of this present work include: (a) attempts to quantitatively evaluate alkyl proton NMR shielding magnitudes in the adducts, relative to maleimide precursors, and (b) use of ab initio Hartree-Fock level calculations to try to reconcile adduct geometries with the observed shielding magnitudes. Our results here complement and extend studies of: (a) adducts of the parent phencyclone with straight-chain N-n-alkylmaleimides, and (b) adducts of 3,6-dibromophencyclone with other symmetrical dienophiles. PMID- 15901323 TI - Subsurface probing in diffusely scattering media using spatially offset Raman spectroscopy. AB - We describe a simple methodology for the effective retrieval of Raman spectra of subsurface layers in diffusely scattering media. The technique is based on the collection of Raman scattered light from surface regions that are laterally offset away from the excitation laser spot on the sample. The Raman spectra obtained in this way exhibit a variation in relative spectral intensities of the surface and subsurface layers of the sample being investigated. The data set is processed using a multivariate data analysis to yield pure Raman spectra of the individual sample layers, providing a method for the effective elimination of surface Raman scatter. The methodology is applicable to the retrieval of pure Raman spectra from depths well in excess of those accessible with conventional confocal microscopy. In this first feasibility study we have differentiated between surface and subsurface Raman signals within a diffusely scattering sample composed of two layers: trans-stilbene powder beneath a 1 mm thick over-layer of PMMA (poly(methyl methacrylate)) powder. The improvement in contrast of the subsurface trans-stilbene layer without numerical processing was 19 times. The potential applications include biomedical subsurface probing of specific tissues through different overlying tissues such as assessment of bone quality through skin, providing an effective noninvasive means of screening for bone degeneration, other skeletal disease diagnosis, and dermatology studies, as well as materials and catalyst research. PMID- 15901324 TI - Role of the micro- and nanostructure in the performance of surface-enhanced Raman scattering substrates assembled from gold nanoparticles. AB - Highly active and stable substrates for surface-enhanced Raman scattering (SERS) can be fabricated by using colloidal crystals to template gold nanoparticles into structured porous films. The structure-dependent performance of these SERS substrates was systematically characterized with cyanide in continuous flow microfluidic chambers. A matrix of experiments was designed to isolate the SERS contributions arising from nano- and microscale porosity, long-range ordering of the micropores, and the thickness of the nanoparticle layer. The SERS results were compared to the substrate structure observed by scanning electron microscopy (SEM) and optical microscopy to correlate substrate structure to SERS performance. The Raman peak intensity was consistently highest for nanoporous substrates with three-dimensionally ordered micropores, and decreases if the micropores are not ordered or not templated. Removing the nanoscale porosity by fusion of the nanoparticles (without removing the large micropores) leads to a drastic plunge in substrate performance. The peak intensity does not strongly correlate to the thickness of the nanoparticle films. The results make possible the efficient controlled fabrication of stable, reproducible, and highly active substrates for SERS based chemical sensors with continuous sampling. PMID- 15901325 TI - Dual layer and multilayer enhancements from silver film over nanostructured surface-enhanced Raman substrates. AB - Novel dual layer and multilayer silver film over nanostructure (SFON) substrates have been developed that provide surface-enhanced Raman scattering (SERS) signal enhancements of greater than 1000% compared to conventional single layer SFON substrates. These substrates provide signal enhancement factors of 3.8 x 10(5) and greater for a variety of SERS active analytes. Substrate preparation is accomplished by vapor depositing a thick (approximately 100 nm) layer of silver on top of an underlying layer of alumina nanoparticles, followed by deposition of additional layers of silver with silver oxide layers between them. Unlike previous dual layer silver island based substrates that have been developed, these substrates do not rely on achieving an optimal morphology via deposition of silver. Instead, these substrates rely on the roughness being provided by the original under-layer, providing enhanced substrate homogeneity and more reproducible signals than either silver island substrates or colloidal substrates. In addition, the signal enhancement gives these substrates extended lifetimes compared to conventional single layer SFON substrates. Finally, this study also shows that geometric surface structure and surface roughness factors play little or no role in this enhancement process, allowing for this multilayer fabrication process to be applied to many different types of substrates achieving similar or even greater results. PMID- 15901326 TI - Surface-enhanced Raman spectra of calf thymus DNA adsorbed on concentrated silver colloid. AB - Raman and surface-enhanced Raman scattering (SERS) spectra of calf thymus DNA were investigated. We have carried out improvements to the silver colloid preparation method of Lee and Meisel in two respects. In one method, the silver sol was boiled with rapid stirring for over two hours. In the second method, the silver sol was concentrated by centrifugation before adding it to the DNA solution. The resulting hydrosol could be stored for 15 months because of its high stabilization. Structural information with respect to the phosphate backbone, deoxyribose, and four bases of DNA could be obtained before and after the DNA solutions were added to the concentrated Ag colloid substrate. The intensities of almost all characteristic bands assigned to various groups of the components of DNA were enhanced to a remarkable degree. The enhancement effect of the DNA solution at neutral pH 7.0 was obviously much better than that at acidic pH 3.4 or at alkaline pH 8.5. Intensity increases of the SERS bands of the DNA solution with time were observed. The SERS signals obtained 16 hours after the interaction of the Ag colloid with the DNA solution were much better than the SERS signals obtained just after the mixed liquid was prepared. This method can be widely used to store the Ag colloid for long times and to obtain the SERS spectra of DNA molecules, and it can further be used to study the adsorption behavior of solute biomacromolecules in different solvents. PMID- 15901327 TI - Real-time imaging of tunable adenosine 5-triphosphate release from an MCM-41-type mesoporous silica nanosphere-based delivery system. AB - We studied a mesoporous silica nanosphere (MSN) material with tunable release capability for drug delivery applications. We employed luciferase chemiluminescence imaging to investigate the kinetics and mechanism of the adenosine 5-triphosphate (ATP) release with various disulfide-reducing agents as uncapping triggers. ATP molecules were encapsulated within the MSNs by immersing dry nanospheres in aqueous solutions of ATP followed by capping of the mesopores with chemically removable caps, such as cadmium sulfide (CdS) nanoparticles and poly(amido amine) dendrimers (PAMAM), via a disulfide linkage. By varying the chemical nature of the ''cap'' and ''trigger'' molecules in our MSN system, we discovered that the release profiles could indeed be regulated in a controllable fashion. PMID- 15901328 TI - On the use of spectroscopic techniques for interaction studies, part I: complexation between europium and small organic ligands. AB - In the framework of environmental studies, it is important to understand the interaction of humic substances with cations (heavy metals, radionuclides) and to determine their complexation constants in order to evaluate their potential impact on their fate. For this purpose, two techniques have been used: electrospray ionization mass spectrometry, a newly used technique in speciation studies, and time-resolved laser-induced fluorescence spectrometry, a well-known technique for such studies. As a first step, for simplification purposes and to compare both techniques, simple molecules having functional groups present in humic substances have been selected, such as acetic, glycolic, and 4 hydroxyphenylacetic acids. Both techniques have been used to obtain stoichiometries and complexation constants between these simple molecules and europium (III). PMID- 15901329 TI - Nanoliter serum sample analysis by mid-infrared spectroscopy for minimally invasive blood-glucose monitoring. AB - The aim of this study was to demonstrate that mid-infrared spectroscopy is able to quantify glucose in a serum matrix with sample volumes well below 1 muL. For this, we applied mid-infrared attenuated total reflectance (ATR) or transmission based spectroscopic methods to glucose quantification in microsamples of dry-film sera, either undiluted or diluted 10 times in distilled water. The sample series spanned physiological glucose concentrations between 50 and 600 mg/dL and volumes of 80, 8, and 1 nL. Calibration was carried out using multivariate partial least squares (PLS) modeling with spectral data between 1180 and 940 cm(-1). Best performance was achieved in the ATR experiments. For raw ATR spectra, the optimum standard error of prediction (SEP) of 13.3 mg/dL was obtained for the 8 nL sample series with subsequent 10-fold dilution. With respect to the coefficient of variation of the glucose assay, CV(pred), we obtained a value of 3% for the 80 nL volume samples with spectral preprocessing using matrix protein absorption bands as an internal standard, 4% for the 8 nL samples, and 6% for the 1 nL samples with raw data. Spectral standardization resulted in significant improvement, especially for the 80 nL volume sample series. By contrast, the accuracy of the glucose assay for the 1 nL sample volume series could not be improved either by internal standardization or by considering the dry film areas for normalization, which we attribute to varying topographies of the dry films. PMID- 15901330 TI - Monitoring the thermal gelation of cellulose ethers in situ using attenuated total reflectance fourier transform infrared spectroscopy. AB - In this work attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy was used to probe the thermal gelation behavior of aqueous solutions of hydroxypropyl methylcellulose (HPMC), specifically thermal gelation and accompanying precipitation. Cloud point measurements are usually evaluated through turbidity in dilute solutions but the method cannot readily be applied to more concentrated or highly viscous solutions. From the ATR-FTIR data, intensity changes of the nu(CO) band marked the onset of gelation and information about the temperature of gelation and the effect of the gel structure on the water hydrogen bonding network was elucidated. Changes in the relative intensities of bands associated with the methoxyl groups and hydrogen-bond-forming secondary alcohol groups indicated that hydrophobic polymer chain interactions were involved in the gelation process. The dominance of inter-molecular H bonding over intra-molecular H bonding within the cellulose ether in solution was also observed. The ATR-FTIR data was in good agreement with measurements of turbidity conducted on the same systems. The work indicates significant potential for the use of ATR-FTIR for the investigation of gelation and cloud point measurements in viscous cellulosic formulations. PMID- 15901331 TI - Mid-infrared fiber-optic attenuated total reflection spectroscopy of the solid liquid phase transition of water. AB - Measurements of mid-infrared (MIR) absorption spectra of water and heavy water were carried out by fiber-optic evanescent wave spectroscopy, using silver halide (AgClBr) infrared fibers. Such measurements were performed for the first time on one sample, during the solid-liquid phase transition. From the variation of the spectra with temperature we found a new isosbestic point (at 3280 cm(-1) for H(2)O or at 2475 cm(-1) for D(2)O) and we identified five components of the O-H (O-D) stretch band. These phenomena have provided new information about the molecular structure of water. PMID- 15901332 TI - A high-sensitivity femtosecond to microsecond time-resolved infrared vibrational spectrometer. AB - We describe an apparatus that provides, for the first time, a seamless bridge between femtosecond and microsecond time-resolved Raman and infrared vibrational spectroscopy. The laser system comprises an actively Q-switched sub-nanosecond pulsed kilohertz laser electronically synchronized to an ultrafast titanium sapphire regenerative amplifier to within 0.2 ns. The ultrafast amplifier provides the stable probe light source enabling high-sensitivity infrared vibrational spectroscopy of transients. Time-resolved infrared spectra of the excited-state relaxation dynamics of metal carbonyl compounds are presented to illustrate the capability of the apparatus, and transient data is resolved from 1 picosecond to over 100 microseconds. The results are compared to conventional nanosecond Fourier transform infrared (FT-IR) and laser based flash photolysis time-resolved infrared technology. PMID- 15901333 TI - Maximum entropy deconvolution of infrared spectra: use of a novel entropy expression without sign restriction. AB - Absorbance and difference infrared spectra are often acquired aiming to characterize protein structure and structural changes of proteins upon ligand binding, as well as for many other chemical and biochemical studies. Their analysis requires as a first step the identification of the component bands (number, position, and area) and as a second step their assignment. The first step of the analysis is challenged by the habitually strong band overlap in infrared spectra. Therefore, it is useful to make use of a mathematical method able to narrow the component bands to the extent to eliminate, or at least reduce, the band overlap. Additionally, to be of general applicability this method should permit negative values for the solution. We present a maximum entropy deconvolution approach for the band-narrowing of absorbance and difference spectra showing the required characteristics, which uses the generalized negative Burg-entropy (Itakura-Saito discrepancy) generalized for difference spectra. We present results on synthetic noisy absorbance and difference spectra, as well as on experimental infrared spectra from the membrane protein bacteriorhodopsin. PMID- 15901334 TI - Comparison of methods for transfer of calibration models in near-infared spectroscopy: a case study based on correcting path length differences using fiber-optic transmittance probes in in-line near-infrared spectroscopy. AB - This article addresses problems related to transfer of calibration models due to variations in distance between the transmittance fiber-optic probes. The data have been generated using a mixture design and measured at five different probe distances. A number of techniques reported in the literature have been compared. These include multiplicative scatter correction (MSC), path length correction (PLC), finite impulse response (FIR), orthogonal signal correction (OSC), piecewise direct standardization (PDS), and robust calibration. The quality of the predictions was expressed in terms of root mean square error of prediction (RMSEP). Robust calibration gave good calibration transfer results, while the other methods did not give acceptable results. PMID- 15901335 TI - Standard reference material 2036 near-infrared reflection wavelength standard. AB - Standard Reference Material 2036 (SRM 2036) is a certified transfer standard intended for the verification and calibration of the wavelength/wavenumber scale of near-infrared (NIR) spectrometers operating in diffuse or trans-reflectance mode. SRM 2036 Near-Infrared Wavelength/Wavenumber Reflection Standard is a combination of a rare earth oxide glass of a composition similar to that of SRM 2035 Near-Infrared Transmission Wavelength/Wavenumber Standard and SRM 2065 Ultraviolet-Visible-Near-Infrared Transmission Wavelength/Wavenumber Standard, but is in physical contact with a piece of sintered poly(tetrafluoroethylene) (PTFE). The combination of glass contacted with a nearly ideal diffusely reflecting backing provides reflection-absorption bands that range from 15% R to 40% R. SRM 2036 is certified for the 10% band fraction air wavelength centroid location, (10%)B, of seven bands spanning the spectral region from 975 nm to 1946 nm. It is also certified for the vacuum wavenumber (10%)B of the same seven bands in the spectral region from 10 300 cm(-1) to 5130 cm(-1) at 8 cm(-1) resolution. Informational values are provided for the locations of thirteen additional bands from 334 nm to 804 nm. PMID- 15901336 TI - Terahertz pulsed spectroscopy as a new tool for measuring the structuring effect of solutes on water. AB - Absorption spectra of aqueous solution of ''chaotropes'' (structure maker) and ''kosmotropes'' (structure breaker) have been recorded in the mid-infrared (MIR) and terahertz (THz) spectral region. A different impact of the two groups of solutes on the absorption spectrum of water was found in the recorded THz spectra. A concentration-dependent increased absorption across the investigated THz spectral region (0.04-2 THz, 1.3-66 cm(-1), respectively) has been recorded for all studied chaotropic solutions, whereas the opposite has been obtained for kosmotrope containing solutions. In the case of ionic solutes a further increase in absorption towards higher frequencies was measured. The distinction between chaotrope and kosmotrope solutes was, as expected, also possible in the MIR spectral region. Depending on the structure-forming effect of the solute the OH stretch vibration of the water (around 3400 cm(-1)) was slightly shifted. A red shift has been observed for solution of kosmotropes, whereas a blue shift was observed in the case of solutions containing chaotropes. Compared to the MIR spectral region the structure influencing effect of solutes can be more efficiently studied in the THz spectral region, which provides information from interactions between neighboring water molecules. PMID- 15901337 TI - Depth profiling of the optical absorption coefficient in ultraviolet-degraded poly(vinylchloride) films by dual beam light profile microscopy. AB - Dual beam laser light profile microscopy (LPM) was applied in this work to the depth mapping of the optical absorption coefficient in photo-degraded poly(vinyl chloride) films. Depth profiles followed the absorption coefficient of a conjugated polyene photoproduct at visible wavelengths in photolyzed films of approximately 200 mum thickness. Both continuous and layered (separable laminate) films were studied. The absorption coefficient profiles reconstructed from photo degraded thin films showed the classic concentration profiles seen in the literature for PVC degraded in the presence of oxygen and nitrogen atmospheres. In the case of single thin layers with continuous properties, the depth profiles were smooth and regular with minimum spatial noise. In the LPM of laminate structures, more optical anomalies were present because of the multiply interfacial structures that appeared in both the images and the reconstructed depth profiles. Notwithstanding, it was possible to profile the optical absorption coefficient at a level of error comparable to standard microtome methods. The latter was determined by comparing the LPM results to a destructive layer-by-layer analysis performed in parallel on the imaged materials. The dual beam LPM method should be generally useful for establishing polyene concentration profiles in industrial materials produced by photochemical, thermal, and chemical degradation mechanisms. PMID- 15901338 TI - Spectroscopic characterization of two different microwave (2.45 GHz) induced argon plasmas at atmospheric pressure. AB - A surface-wave-sustained discharge created by using a surfatron device in a tube open to the atmosphere can be used to maintain a microwave (2.45 GHz) plasma at atmospheric pressure at powers of less than 300 W. The TIA (Torche a Injection Axiale) is a device also producing a plasma that, moreover, permits us to work at high power (higher than 200 W and up to 1000 W). A study of the departure from the thermodynamic equilibrium existing in the argon plasmas created by both devices has been done by using optical emission spectroscopy techniques in order to characterize them and to evaluate their possible advantages when they are used for applied purposes. PMID- 15901339 TI - Investigation of the state of local thermodynamic equilibrium of a laser-produced aluminum plasma. AB - In this work, the assumption of local thermodynamic equilibrium (LTE) for a laser induced plasma in ambient air is examined experimentally using two different laser systems, namely an infrared short-pulse Ti : Sapphire laser and an ultraviolet long-pulse XeCl excimer laser. The LTE assumption is investigated by examining the plasma produced at a laser fluence of 10 J/cm(2) from aluminum targets containing iron and magnesium impurities. The excitation temperature is deduced from Boltzmann diagrams built from a large number of spatially integrated neutral iron lines distributed from 3.21 to 6.56 eV. It is shown that at any time after the end of the laser pulse, the neutral excited states are in excellent Boltzmann equilibrium. Detailed investigation of Boltzmann equilibrium further validates previous temperature measurements using less accurate diagrams. However, observations of ion lines provide some evidence that the ionized species do not obey Saha equilibrium, thereby indicating departure from LTE. This could be explained by the fact that the plasma cannot be considered as stationary for these species. PMID- 15901340 TI - Characterization of delamination cracks in Al2O3/Al2O3+3Y-TZP multilayered composite by raman and fluorescence piezo-spectroscopy. PMID- 15901341 TI - A simple approach to normalization for spectroscopic data mining. PMID- 15901342 TI - The use of hypromellose in oral drug delivery. AB - Hypromellose, formerly known as hydroxypropylmethylcellulose (HPMC), is by far the most commonly employed cellulose ether used in the fabrication of hydrophilic matrices. Hypromellose provides the release of a drug in a controlled manner, effectively increasing the duration of release of a drug to prolong its therapeutic effect. This review provides a current insight into hypromellose and its applicability to hydrophilic matrices in order to highlight the basic parameters that affect its performance. Topics covered include the chemical, thermal and mechanical properties of hypromellose, hydration of the polymer matrices, the mechanism of drug release and the influence of tablet geometry on drug-release rate. The inclusion of drug-release modifiers within hypromellose matrices, the effects of dissolution media and the influence of both the external environment and microenvironment pH within the gel matrix on the properties of the polymer are also discussed. PMID- 15901343 TI - Multidrug resistance modulator interactions with neutral and anionic liposomes: membrane binding affinity and membrane perturbing activity. AB - A variety of cationic lipophilic compounds (modulators) have been found to reverse the multidrug resistance of cancer cells. In order to determine the membrane perturbing efficacy and the binding affinity of such drugs in neutral and anionic liposomes, the leakage of Sulfan blue induced by five modulators bearing different electric charges was quantified using liposomes with and without phosphatidic acid (xEPA=0 and 0.1), at four lipid concentrations. The binding isotherms were drawn up using the indirect method based on the dependency of the leakage rate on the modulator and the lipid concentrations. Upon inclusion of negatively charged lipids in the liposomes: (i) the binding of cationic drugs was favoured, except in a case where modulator aggregation occurred in the lipid phase; (ii) the drugs with a net electric charge greater than 1.1 displayed a greater enhancement in their potency to produce membrane perturbation; and (iii) the EPA effect on membrane permeation was due mainly to that on membrane perturbation (>or=50%) and, to a lesser extent, to that on the binding affinity (or=3.4 L kg-1) and half-life (t1/2; 7 h), negligible plasma protein binding (approximately 0.1%), low/intermediate oral uptake (>or=13% as intact substance) and low and varying oral bioavailability (mean 1.4% in minipigs and 3.9% in dogs). Following administration of therapeutically relevant oral doses, plasma concentrations of AZD3582 were very low (40 h in rats, minipigs and dogs, respectively. The Vss and CL for naproxen were small. Plasma protein binding was extensive, and saturation was observed within the therapeutic dose and concentration range. Intake of food prolonged the systemic absorption of naproxen in the minipig. The pharmacokinetics of naproxen did not show apparent time- or gender-related dependency. Following oral dosing of [3H]-, [14C]- and [15N]-AZD3582, most [14C]- and [3H]-activity was excreted in urine and expired air, respectively. Seventeen per cent of [15N] was recovered in minipig urine as [15N]-nitrate. About 30% of [3H]-activity (naproxen and/or naproxen-related metabolites) was excreted in bile and re-absorbed. Concentrations of [14C]-activity (nitrooxy-butyl group and/or its metabolites) in milk were higher than in plasma and [3H]-activity in milk. [3H]- and [14C]-excretion data indicated that intact AZD3582 was not excreted in urine, bile or milk to a significant extent. There was no apparent consistency between tissue distribution of [14C]- and [3H]-activity in the rat, which suggests rapid and extensive metabolism of extravascularly distributed AZD3582. A substantial increase of plasma nitrate levels was found after single and repeated oral doses of AZD3582 in the minipig. No inhibition or induction of CYP450 was found. PMID- 15901349 TI - S-adenosyl-L-methionine: transcellular transport and uptake by Caco-2 cells and hepatocytes. AB - S-adenosyl-L-methionine (SAMe) is an endogenous molecule that is known to be protective against hepatotoxic injury. Although oral SAMe appears to be absorbed across the intestinal mucosa, its systemic bioavailability is low. The reason for this is unknown. Using the Caco-2 cell culture model for enterocyte absorption, we determined the mode by which SAMe is transported across this cell monolayer. We also determined the extent it is taken up by both Caco-2 cells and hepatocytes. In Caco-2 cells transport was observed in both apical to basolateral and basolateral to apical directions. The apparent permeability coefficients (Papp) appeared to be concentration independent and were similar in both directions (0.7x10(-6) and 0.6x10(-6) cm s-1, respectively), i.e. identical to that of the paracellular transport marker mannitol (0.9x10(-6) and 0.7x10(-6) cm s-1). This mode of transport was supported by a four-fold increase in the Papp for SAMe transport in Ca++-free buffer. Cellular uptake of SAMe was examined in both Caco-2 cells and cultured rat hepatocytes. Uptake by hepatocytes was not saturable in a concentration range of 0.001-100 microM. Accumulation by both cell types was very low, with a cell:medium ratio at equilibrium of only 0.2-0.5. This low cell accumulation supports the finding of paracellular transport as the only mode of cell membrane transport. Increased hepatocellular protection for SAMe may be accomplished by converting SAMe to a more lipid-soluble prodrug. PMID- 15901350 TI - In-vitro and in-vivo anti-inflammatory and antinociceptive effects of the methanol extract of the roots of Morinda officinalis. AB - The anti-inflammatory effects of the methanol extract of the roots of Morinda officinalis (MEMO) (Rubiaceae) were evaluated in-vitro and in-vivo. The effects of MEMO on lipopolysaccharide (LPS)induced responses in the murine macrophage cell line RAW 264.7 were examined. MEMO potently inhibited the production of nitric oxide (NO), prostaglandin E2 and tumour necrosis factor-alpha (TNF-alpha) in LPS-stimulated RAW 264.7 macrophages. Consistent with these results, the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein level, and of iNOS, COX-2 and TNF-alpha at the mRNA level, was also inhibited by MEMO in a concentration-dependent manner. Furthermore, MEMO inhibited the nuclear factor kappa B (NF-kappaB) activation induced by LPS, and this was associated with the prevention of degradation of the inhibitor kappaB (IkappaB), and subsequently with attenuated p65 protein in the nucleus. The anti inflammatory effect of MEMO was examined in rats using the carrageenan-induced oedema model. The antinociceptive effects of MEMO were assessed in mice using the acetic acid-induced abdominal constriction test and the hot-plate test. MEMO (100, 200 mg kg-1 per day, p.o.) exhibited anti-inflammatory and antinociceptive effects in these animal models. Taken together, the data demonstrate that MEMO has anti-inflammatory and antinociceptive activity, inhibiting iNOS, COX-2 and TNF-alpha expression by down-regulating NF-kappaB binding activity. PMID- 15901351 TI - Protective effect of aqueous garlic extract against naphthalene-induced oxidative stress in mice. AB - The aim of this study was to investigate the possible protective effects of aqueous garlic extract (AGE) against naphthalene-induced oxidative changes in liver, kidney, lung and brain of mice. Balb/c mice (25-30 g) of either sex were divided into five groups each comprising 10 animals. Mice received for 30 days: 0.9% NaCl, i.p. (control); corn oil, i.p; AGE in a dose of 125 mg kg-1, i.p.; naphthalene in a dose of 100 mg kg-1, i.p. (dissolved in corn oil); and AGE (in a dose of 125 mg kg-1, i.p.) plus naphthalene (in a dose of 100 mg kg-1, i.p.). After decapitation, liver, kidney, lung and brain tissues were excised. Malondialdehyde (MDA) and glutathione (GSH) levels and myeloperoxidase activity (MPO) were determined in the tissues, while oxidant-induced tissue fibrosis was determined by collagen content. Tissues were also examined microscopically. Serum aspartate aminotransferase, alanine aminotransferase levels and blood urea nitrogen and creatinine concentrations were measured for the evaluation of hepatic and renal function, respectively. MDA and GSH levels were also assayed in serum samples. In the naphthalene-treated group, GSH levels decreased significantly, while MDA levels, MPO activity and collagen content increased in the tissues (P<0.01-0.001), suggesting oxidative organ damage, which was also verified histologically. In the AGE-treated naphthalene group, all of these oxidant responses were reversed significantly (P<0.05-0.01). Hepatic and renal function test parameters, which increased significantly (P<0.001) following naphthalene administration, decreased (P<0.05-0.001) after AGE treatment. The results demonstrate the role of oxidative mechanisms in naphthalene-induced tissue damage. The antioxidant properties of AGE ameliorated oxidative organ injury due to naphthalene toxicity. PMID- 15901352 TI - Isobolographic analysis of the sedative interaction between six central nervous system depressant drugs and Valeriana edulis hydroalcoholic extract in mice. AB - It has been declared frequently that valerian may potentiate the effect of other central nervous system (CNS) depressant drugs, however there has been a lack of experimental data. We have evaluated the profile of the interactions between the ethanol extract of Valeriana edulis spp procera and six CNS depressant drugs using an exploratory model to test the sedative effect in mice. All the compounds tested showed a dose-dependent sedative effect with the following ED50 values: valerian 181.62, diazepam 1.21, ethanol 1938, pentobarbital 11.86, buspirone 1.04, haloperidol 0.41 and diphenhydramine 17.06 mg kg-1. An isobolographic analysis was used to evaluate the sedative interaction of the intraperitoneal co administration of 1:1 fixed-ratio combination of equi-effective doses of valerian extract with each CNS depressant drug. The ED50 theoretical (Zadd) and experimental (Zexp) for each combination were: valerian+diazepam,Zadd=91.41 mg kg 1, Zexp=81.64 mg kg-1; valerian+ethanol, Zadd=1060.22 mg kg-1, Zexp=687.89 mg kg 1; valerian+pentobarbital, Zadd=96.74 mg kg-1, Zexp=151.83 mg kg-1; valerian+buspirone, Zadd=91.33 mg kg-1, Zexp=112.73 mg kg-1; valerian+haloperidol, Zadd=91.01 mg kg-1, Zexp=91.52 mg kg-1; valerian+diphenhydramine, Zadd=99.34 mg kg-1, Zexp=123.52 mg kg-1. Neither synergistic nor attenuate effects were found in any of the combinations evaluated. We concluded that the valerian extract did not potentiate the sedative effect of commonly prescribed CNS depressant drugs as was expected. The additive effect found through the isobolographic analysis suggested that the sedative effect of V. edulis resulted from the activation of common mechanisms of haloperidol, diazepam, buspirone, pentobarbital, diphenhydramine and ethanol. PMID- 15901353 TI - Influence of the Ginkgo extract EGb 761 on rat liver cytochrome P450 and steroid metabolism and excretion in rats and man. AB - Extracts from leaves of Ginkgo biloba L. are among the most used herbal medicinal products worldwide. Based on in-vitro tests and studies in rats, concern has been expressed that intake of Ginkgo extracts may affect hepatic metabolism of xenobiotics and cause drug interactions, although no evidence for modulation of cytochrome P450 (CYP450) enzyme activity was obtained in human trials. Because of these contradictory findings, we investigated the effects of the standardised extract EGb 761 on hepatic CYP450 in rats. EGb 761 (100 mg kg-1 daily, p.o., for 4 days) strongly increased liver CYP450 content and altered the ex-vivo biotransformation of androstendione, as well as metabolism of endogenous steroids. However, in human subjects no effect on the urinary steroid profile was observed after intake of EGb 761 for 28 days (240 mg daily). These results indicate that the effects of EGb 761 on drug metabolising enzymes are specific for rats and may not be extrapolated to man. PMID- 15901354 TI - Nelumbinis Semen reverses a decrease in hippocampal 5-HT release induced by chronic mild stress in rats. AB - Depression is associated with a dysfunctional serotonin system. Recently, several lines of evidence have suggested that a very important evoking factor in depression may be a serotonin deficit in the hippocampus. This study assessed the antidepression effects of Nelumbinis Semen (NS) through increasing serotonin concentrations under normal conditions and reversing a decrease in serotonin concentrations in rat hippo-campus with depression-like symptoms induced by chronic mild stress (CMS). Using an in-vivo microdialysis technique, the serotonin-enhancing effect of NS on rat hippocampus was investigated and its effects compared with those of two well-known antidepressants, Hypericum perforatum (St John's wort) and fluoxetine (Prozac). Rats were divided into five groups: saline-treated normal, without CMS; saline-treated stress control; NS-, St John's wort- and fluoxetine-treated rats under CMS for 8 weeks or no stress treatment. NS and fluoxetine significantly increased serotonin in normal conditions and reversed a CMS-induced decrease in serotonin release in the hippocampus (P<0.05 compared with normal group or control group under CMS). These results suggest that NS increases the serotonin levels normally decreased in depression, resulting in an enhancement of central serotonergic transmission and possible therapeutic action in depression. It is suggested that NS may present an antidepressant effect through enhancement of serotonin. PMID- 15901355 TI - Effect of the anticancer drug oracin on mouse liver topoisomerases I and II. AB - The inhibitory effect of oracin on the activities of topoisomerases I and II isolated from the nuclei of mice liver tissue was studied. This drug showed a 100% inhibitory effect at 5 microM and 50% inhibition at 1 microM on topoisomerase II activity, while the activity of topoisomerase I at these concentrations was not inhibited. PMID- 15901356 TI - Kinetic characterization of glycosidase activity from disaccharide conjugate to monosaccharide conjugate in Caco-2 cells. AB - Glycosidase activity influences the intestinal absorption of glycosides. Our previous study in rats suggested that disaccharide conjugates might be prototypes for pre-prodrugs aiming at the Na(+)/glucose co-transporter-mediated transport of prodrugs (drug glucoside) as a novel absorption pathway. One of the crucial factors is the formation of a glucoside drug from the disaccharide conjugate. Since there is a large species difference in metabolism, it is necessary to examine the cells and/or enzymes derived from human tissue to confirm this concept. In this paper, we kinetically characterized the glycosidase activity of disaccharide conjugates in Caco-2 cells. Disaccharide conjugates of p-nitrophenol (p-NP) (p-NP beta-cellobioside, p-NP beta-lactoside and p-NP beta-maltoside) were hydrolysed to p-NP beta-glucoside. beta-glucosidase or beta-galactosidase (lactase/phloridzin hydrolase, LPH) and alpha-glucosidase (sucrase-isomaltase) had different pH-dependent activities for disaccharide conjugates. At neutral pH, LPH has low affinity and low capacity, and sucrase-isomaltase has high affinity and high capacity, whereas at acid pH, LPH has high affinity and low capacity, and sucrase-isomaltase has low affinity and high capacity. The hydrolysis clearance calculated with Vmax/Km indicated that sucrase-isomaltase activity is much higher than LPH activity at either neutral or acid pH in Caco-2 cells. Since the hydrolysis rate of the disaccharide conjugate was highly dependent on the pH value and type of glycoside linkage, the appropriate selection of a glycoside form after consideration of these differences is the key to designing a sugar conjugate prodrug. PMID- 15901357 TI - Improvement of diabetic states in streptozotocin-induced type 1 diabetic rats by vanadyl sulfate in enteric-coated capsules. AB - Chronic oral administration of vanadyl sulfate has recently been shown to improve the state of type 2 diabetic subjects. Mild gastrointestinal symptoms and side effects, however, have been observed in some subjects. To find safer and more effective dosages, we have developed an enteric-coated capsule containing solid vanadyl sulfate (ECC/VS), which enhances the bioavailability of vanadyl sulfate to almost double that of vanadyl sulfate solution. ECC/VS was chronically administered to treat streptozotocin-induced diabetic rats (STZ-rats), an animal model of type 1 diabetes mellitus, and an equivalent blood-glucose-lowering effect was observed at half the doses of vanadyl sulfate alone. In addition, we observed almost the same total vanadium levels in the serum after chronic administration of ECC/VS as those of vanadyl sulfate alone, suggesting that plasma vanadium levels correlate with the hypoglycaemic activity of vanadyl sulfate. These results indicate that oral ECC/VS improves the diabetic state by enhancing the uptake of vanadium in STZ-rats. These findings will be useful in designing clinical trials of vanadyl sulfate for diabetic subjects. PMID- 15901358 TI - Carbon dioxide for euthanasia: concerns regarding pain and distress, with special reference to mice and rats. AB - Carbon dioxide (CO2) is the most commonly used agent for euthanasia of laboratory rodents, used on an estimated tens of millions of laboratory rodents per year worldwide, yet there is a growing body of evidence indicating that exposure to CO2 causes more than momentary pain and distress in these and other animals. We reviewed the available literature on the use of CO2 for euthanasia (as well as anaesthesia) and also informally canvassed laboratory animal personnel for their opinions regarding this topic. Our review addresses key issues such as CO2 flow rate and final concentration, presence of oxygen, and prefilled chambers (the animal is added to the chamber once a predetermined concentration and flow rate have been reached) versus gradual induction (the animal is put into an empty chamber and the gas agent(s) is gradually introduced at a fixed rate). Internationally, animal research standards specify that any procedure that would cause pain or distress in humans should be assumed to do so in non-human animals as well (Public Health Service 1986, US Department of Agriculture 1997, Organization for Economic Cooperation and Development 2000). European Union guidelines, however, specify a certain threshold of pain or distress, such as 'skilled insertion of a hypodermic needle', as the starting point at which regulation of the use of animals in experimental or other scientific procedures begins (Biotechnology Regulatory Atlas n.d.). There is clear evidence in the human literature that CO2 exposure is painful and distressful, while the non human literature is equivocal. However, the fact that a number of studies do conclude that CO2 causes pain and distress in animals indicates a need for careful reconsideration of its use. Finally, this review offers recommendations for alternatives to the use of CO2 as a euthanasia agent. PMID- 15901359 TI - A simple method for assessing muscle function in common marmosets. AB - A novel method of assessing muscle function in the common marmoset was developed as part of a multidisciplinary long-term study. The method involved home cage presentation of a weight-pulling task. Over a 4-5 month period, 38 of 42 animals were successfully trained to displace weights of up to 920 g (mean 612+/-20 g). Performance, following initial training, was stable and independent of gender or body weight. PMID- 15901360 TI - A novel method for activity monitoring in small non-human primates. AB - Patterns of spontaneous activity are valuable reflections of well-being in animals and humans and, because of this, investigations have frequently incorporated some form of activity monitoring into their studies. It is widely believed that activity monitoring, alongside assessments of general behaviour, should be included in initial CNS safety pharmacology screening. As the number of marmoset studies having actimetry as their focus, or as an adjunct, is increasing, we wished to evaluate an alternative approach to those commonly used. The method is based on miniaturized accelerometer technologies, currently used for human activity monitoring.Actiwatch-Minis were used to monitor the activity of two groups of differently housed marmosets for 14 consecutive days. Group A consisted of four mixed-sex pairs of animals and group B comprised eight group housed males. Activity profiles were generated for weekday and weekend periods. The devices captured quantifiable data which showed differences in total activity between the two differently housed groups and revealed intragroup variations in the temporal spread of activity between weekdays and weekends. The Actiwatch-Mini has been shown to generate retrospective, data-logged activity counts recorded from multiple animals in a single arena by means of non-invasive monitoring. PMID- 15901361 TI - The isolated working heart model in infarcted rat hearts. AB - Congestive heart failure (CHF) is one of the most common causes of death in western countries. The aim of this study was to establish and validate the working heart model in rat hearts with CHF. In the rat model the animals show parameters and symptoms that can be extrapolated to the clinical situation of patients with end-stage heart failure. The focus of attention was the evaluation of cardiodynamics (e.g.contractility) in the isolated 'working heart' model. The geometric properties of the left ventricle were measured by planimetry (stereology). Formulae available in the past for determining certain parameters in the working heart model (e.g.external heart work) have to be fitted to the circumstances of the infarcted rat hearts with its different organ properties.CHF was induced in Wistar Kyoto (WKY/NHsd) and spontaneously hypertensive rats (SHR/NHsd) by creating a permanent (8 week) occlusion of the left coronary artery, 2 mm distal to the origin from the aorta, by a modified technique (Itter et al. 2004). This resulted in a large infarction of the free left ventricular wall. We were able to establish and adapt a new and predictive working heart model in spontaneously hypertensive rat hearts with myocardial infarction (MI) 8 12 weeks after coronary artery ligation. At this stage the WKY rat did not show any symptoms of CHF. The SHR rat represented characteristic parameters and symptoms that could be extrapolated to the clinical situation of patients with end-stage heart failure (NYHA III-IV). Upon inspection, severe clinical symptoms of CHF such as dyspnoea, subcutaneous oedema, palebluish limbs and impaired motion were prominent. On necropsy the SHR showed lung oedema, hydrothorax, large dilated left and right ventricular chambers and hypertrophy of the septum. In the working heart model the infarcted animals showed reduced heart power, diminished contractility and enhanced heart work, much more so in the SHR/NHsd than in the Wistar Kyoto rat (WKY/NHsd). The aim for the future is to find a causal therapy of heart failure treatment. At present, only palliative therapy is possible for patients with heart failure. For this reason the working heart model in CHF rat hearts should provide a valuable method for early testing of new therapeutic approaches for patients with CHF. PMID- 15901362 TI - Development of a gut perfusion model as an alternative to the use of live fish. AB - An alternative fish model with the principal aim of studying the interaction between fish pathogens and the intestinal tissue was developed. The preparation consisted of an excised gut tractus from rainbow trout (Oncorhynchus mykiss), perfused through cannulation of the aorta intestinalis ventralis with filtered and heparinized Cortland+dextran 1% as the perfusion fluid. This perfusion fluid was delivered by means of a drip. The perfused gut tractus was suspended in a circular bath filled with Ringer solution, which was aerated and kept at a constant temperature of 12 degrees C. Unperfused gut placed in Ringer solution at the same temperature served as the negative control. Perfusion was effective in maintaining the gut in a healthy condition for at least 60 min with only slight oedema and sloughing of the epithelium. Conversely, the unperfused gut revealed excessive tissue degeneration and severe necrosis. PMID- 15901363 TI - Indicators for post-surgery recovery in Mongolian gerbils (Meriones unguiculatus). AB - Telemetric investigations of various parameters are widely used to estimate an animal's state. However, the implantation of the transmitters includes anaesthesia and surgery and has short and longer lasting impacts on the studied object. The aim of the present paper was to evaluate these effects in Mongolian gerbils, namely the hypothermia caused by the anaesthetic and the enduring disturbance of daily rhythmicity until complete recovery. The surgery associated with the implantation of the transmitters differed both in severity and type of anaesthesia. Whereas normal values of body temperature were restored within hours, restoration of daily rhythm required several days, depending on the severity of the surgical procedure. Also, the sensitivity of the body temperature to activity changes was different until the rhythms were re-established. A method based on the rhythm magnitude and shape was proposed to estimate the time until complete recovery of the animals. PMID- 15901364 TI - Quantitative assessment of mouse skin transplant rejection using digital photography. AB - Mouse skin transplantation is an established in vivo model used to investigate the T-cell-mediated immune response of acute allograft rejection. The critical endpoint of this model is complete rejection of the allograft. However, visual judgement of this end stage of rejection is an arbitrary process and difficult to standardize. To overcome this problem, we established a monitoring method based on digital photography. Serial pictures from skin allografts of individual animals (C57BL/6 on BALB/c) were taken with a digital camera mounted on a microsurgical microscope. Thereby, the description and the correct assessment of early stages of rejection were possible due to the magnification of grafts by the microscope. Rejection scores were introduced to describe different stages from retained to completely rejected grafts. With cyclosporine A as a standard immunosuppressive treatment, we showed that early stages of skin rejection were unambiguously identified. This procedure allows the earlier termination of the experiment and reduction of animal distress, and it can be re-evaluated anywhere and any time after completion.This study demonstrates the suitability of monitoring experimental skin allograft rejection by digital photography, entailing several refinements in animal experimentation, both for the researcher and for the animal. PMID- 15901365 TI - Biopsy of the mouse prostate. AB - Many transgenic and knockout mouse models of prostate cancer have become available over the past decade. In this paper we describe a simple biopsy technique of the murine prostate. This technique allows sequential follow-up of the prostate in an individual mouse. Its use could also reduce the number of mice used in studies of the prostate gland. PMID- 15901366 TI - Multiple in vivo passages enhance the ability of a clinical Helicobacter pylori isolate to colonize the stomach of Mongolian gerbils and to induce gastritis. AB - The Mongolian gerbil is an excellent animal model for Helicobacter pylori-induced gastritis in humans. In this study, initially low colonization rates of the H. pylori strains ATCC 43504, SS1, or HP87 inoculated into gerbils caused difficulties in establishing this model. In order to increase the colonization ability and pathogenicity, the clinical HP87 isolate was selected for adaptation to the gerbil stomach by multiple in vivo passages through gerbils. Development of gastritis was examined histologically at 4-52 weeks after infection. The proportion of gerbils which tested positive for H. pylori by culture at four weeks after inoculation gradually increased from 11.1% of gerbils inoculated with HP87 without prior in vivo passage (P0) to 100% of gerbils inoculated with HP87 with seven in vivo passages (P7). In addition, adaptation of HP87 resulted in more severe histopathological changes. Gerbils infected with adapted HP87 (P7) exhibited severe infiltration by monomorphonuclear and polymorphonuclear leukocytes in the mucosa, submucosa, and subserosa of the gastric antrum, as well as epithelial changes consisting of hyperplasia, erosion, and ulceration. Histopathological changes increased in severity from four to 52 weeks after infection. Adaptation of HP87 during its passages through gerbils could be due to genetic changes in bacterial colonization factors. Identification of these changes might be useful to understand the underlying mechanism of gastric adaptation and pathogenesis of H. pylori. PMID- 15901367 TI - Mycotoxins in laboratory rodent feed. AB - Twenty-one batches of fixed-formula rodent diets from three feed manufacturers were tested for the presence of five mycotoxins: deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin, T-2 toxin and ochratoxin A (OTA). Five batches were also tested for the presence of zearalenone (ZEN) and six batches for aflatoxins. Detectable levels of DON (up to 298 microg/kg), NIV (up to 118 microg/kg), OTA (up to 3.1 microg/kg) or ZEN (up to 26.7 microg/kg) were found in samples from all manufacturers. Three batches contained two (DON or NIV and OTA or ZEN) and one batch contained three (DON, OTA and ZEN) different mycotoxins. Aflatoxins, T 2 and HT-2 were not detected in any of the batches. The concentrations of mycotoxins detected in the feed were low, but indicated that feed ingredients, probably the cereal ingredients, were contaminated by mycotoxins. Since mycotoxins are known to have toxic and/or immunosuppressive effects, non contaminated ingredients should be used for production of laboratory animal feed. The results imply that an improved quality control of ingredients used for laboratory rodent feed should be implemented. PMID- 15901368 TI - Blood sampling from the cranial vena cava in the Norway rat (Rattus norvegicus). AB - This paper describes blood sampling from the cranial vena cava (CVC) in the Norway rat. In order to limit stress, the blood sampling should be done under short-term inhalation anaesthesia, for example, an oxygen/isoflurane mixture. The injection site is just cranial to the first rib, 0.3-0.8 cm lateral to the manubrium when the animal is in dorsal recumbency. The needle, attached to a syringe, is inserted at 30 degrees in the direction of the opposite femoral head. After penetration of the skin, negative pressure is developed in the syringe and the insertion of the needle is continued for another 0.2-1 cm in the given direction until blood begins to flow. The amount of blood sampled ranges from 0.8 to 2.5 mL depending on the body weight of the patient. A trial on 50 rats aged 5 24 months included 25 rats sampled once, eight rats sampled twice with an interval of seven days, 11 rats sampled twice with an interval of three weeks and four rats sampled four times with intervals of four weeks--a total of 87 blood samplings. The serious complications quoted in association with blood sampling from the CVC in other experimental animals (vascular lacerations, heart puncture, serious haemorrhage, tracheal and throat trauma) were not observed in our study. There were only four blood samplings (4.5%) with mild haemorrhage from the injection site, due to erroneous sampling from the jugular vein. PMID- 15901369 TI - Effect of accommodating sucking and nosing on the behaviour of artificially reared piglets. AB - Neonatal piglets are often used in biomedical research applications that require artificial rearing. Social housing can be problematic because the piglets develop belly nosing, navel and ear sucking that can result in injury. Our objective was to determine the effectiveness of using feeding devices that provide various opportunities for sucking and nosing behaviour on reducing piglet-directed behaviour of group-housed laboratory piglets. Fifteen piglets were used in each of four trials. The piglets nursed their dam for approximately 72 h to obtain passive immunity before transfer to a laboratory facility where they were allotted, five per group, to one of three stainless steel isolator units. Each unit featured a different style of feeding system for the delivery of milk replacer: a plastic trough (T), a nipple (N) mounted on a smooth plexiglass wall, or a nipple mounted on a pliant bag of sterile water (artificial udder [AU]). Each system had five feeding spaces so that all piglets fed simultaneously. Milk was provided at 6-h intervals, and behaviour was recorded on alternate days for 12 days post-weaning. Although trough-fed piglets began to eat much sooner than those piglets fed from nipples, time spent nosing, chewing or sucking on pen mates and belly nosing were markedly higher in T piglets than in either N or AU, overall (mean: P<0.05) and over time (quadratic: P<0.05). Over time, N piglets developed a stereotypic snout rubbing on the wall behind the nipples, while AU piglets massaged and often fell asleep in contact with the udder from day 2 of the trial. Resting patterns were also affected. N and AU piglets settled down to rest more quickly (P<0.01) and spent significantly more time resting in the hour following feeding than T piglets (P<0.05). A feeding device that accommodates both sucking and massage can significantly reduce piglet-directed behaviour and may facilitate social housing of artificially reared piglets. PMID- 15901372 TI - Hunterian Lecture. Regulation of hypoxia-inducible factor 1 (HIF-1) by nitric oxide in oral squamous cell carcinoma. AB - INTRODUCTION: Hypoxia is a common feature of many cancers, contributing to tumour progression as well as potentially compromising radiotherapy and chemotherapy. Hypoxia-inducible factor 1 (HIF-1) is an essential component in changing the transcriptional response of tumours under hypoxia and it targets the transcription of many genes involved in cancer biology. Over-expression of HIF-1 has been associated with increased patient mortality in several cancer types. Regulation of HIF-1 by the signalling molecule nitric oxide (NO) is becoming increasingly recognised. METHOD: Three oral cancer cell lines were used to investigate the effects of NO synthase enzymes (NOS) on HIF-1alpha expression under both normal oxygen and hypoxic conditions. The effect of NOS inhibition was evaluated with the drug L-NMMA. Protein expression was determined with Western blotting. RESULTS & CONCLUSIONS: HIF-1alpha expression occurred following exposure of cells to 3% oxygen for 8 h in all three cell lines, and its expression was found to be dependent on NOS activity, being reduced or inhibited by L-NMMA. Although the mechanism remains to be established, NO appears to play a role in the expression of HIF-1alpha in oral cancer. The possible clinical implications of targeting HIF-1 in cancer are discussed. PMID- 15901373 TI - The case for radical prostatectomy. PMID- 15901374 TI - Pancreatic cancer--is an aggressive approach justified? AB - INTRODUCTION: Surgery is the only curative treatment for carcinoma of the pancreas. Resection rates can be low (4.5%), figures of 30% have also been suggested as possible. The approach undertaken in this unit is to consider all patients as potentially resectable unless otherwise proven. PATIENTS & METHODS: 140 patients were studied over 6-year period; 113 underwent palliative treatment (48% distant metastases, 40% local spread, 11% high operative risk); 14 had a triple bypass (14/113 = 12%), 99 were managed conservatively, 43 received palliative chemotherapy. 23/140 (16%) underwent Whipple's procedure (n = 23; 12 females, 11 males; mean age, 60 years); 4/23 had chronic pancreatitis. Distal pancreatectomy was undertaken in 4 patients. RESULTS: Median survival time for patients undergoing a triple bypass was 5 months (range, 0.1-20 months), 3 months for patients treated conservatively (range, 0.1-30 months) and 5 months for patients undergoing palliative chemotherapy (range, 1-30 months). 30-day mortality for Whipple's procedure was 4% (1/23) with median survival rate for patients with carcinoma of 13 months (range, 5-66 months); 31 months for patients with clear resection margins and negative nodes (n = 5). CONCLUSION: This policy allows a resection rate of 19% with increased median survival rate for patients with cancer by 8 months more than those who where not resected. Aggressive staging and pancreatic biopsies allow patients to be entered into chemotherapy trials with improvement in survival and potential future benefits. PMID- 15901375 TI - Hepatic resection for breast cancer metastases. AB - INTRODUCTION: Hepatic resection is an established modality of treatment for colorectal cancer metastases. Resection of breast cancer liver metastases remains controversial, but has been shown to be an effective treatment in selected cases. This study reports the outcome of 8 patients with liver metastases from breast cancer. PATIENTS & METHODS: 8 patients with liver metastases from previously treated breast cancer were referred for hepatic resection between September 1996 and December 2002. Six were eligible for liver resection. The mean age was 45.8 years. The resections performed included 1 segmentectomy and 5 hemihepatectomies of which one was an extended hemihepatectomy. One patient had a repeat hepatectomy 44 months after the first resection. RESULTS: There were no postoperative deaths or major morbidity. The resectability rate was 75%. Follow up periods range from 6 to 70 months with a median survival of 31 months following resection. There have been 2 deaths, one died of recurrence in the residual liver at 6 months and one died disease-free from a stroke. Of the remaining 4 patients, 1 has had a further liver resection at 44 months following which she is alive and 'disease-free' at 70 months. The one patient with peritoneal recurrence is alive 49 months after her liver resection with 2 patients remaining disease-free. CONCLUSION: Hepatic resection for breast cancer liver metastases is a safe procedure with low morbidity and mortality. PMID- 15901376 TI - Soft tissue sarcomas: are current referral guidelines sufficient? AB - INTRODUCTION: To investigate the adequacy of current early referral guidelines for patients with extremity soft tissue sarcomas. PATIENTS & METHODS: 365 patients with confirmed soft tissue sarcomas were evaluated. Data were collected prospectively and included the length of history and the presence of features in current guidelines suggestive of malignancy (pain, rapidity of growth, depth and tumour size). Statistical analysis included the t-test, ANOVA and the Chi test. RESULTS: Deep tumours were the commonest (306 patients with deep tumours). Pain was the least consistent feature (176 patients with pain). 345 patients with one or more of the guideline features had an average history of 19.86 months, 238 of these were seen after more than 3 months. CONCLUSION: Although the majority of soft tissue sarcomas in our patients had one or more of the clinical guideline features, there was still an unacceptable delay in referring these patients to a specialist unit. The referral guidelines should be modified with special emphasis on depth, which is the most sensitive, followed by size and a history of rapid growth. This combined with increased awareness of these guidelines and a well advertised, open-access clinic linked to a specialist unit should allow for a more rapid evaluation of soft tissue tumours. PMID- 15901377 TI - A team approach to musculo-skeletal disorders. AB - INTRODUCTION: The majority of patients with musculo-skeletal problems referred to hospitals in the UK have to wait for months, if not over a year, before finally seeing an orthopaedic surgeon. In Stobhill Hospital, Glasgow, the waiting time for an out-patient appointment was 182 days in 1995, with only 20% of the referrals requiring surgery. The aim of this paper was to reduce the out-patient waiting times based on a co-ordinated team approach. METHODS: An outpatient musculo-skeletal service was developed over a 7-year period at Stobhill Hospital. The traditional consultant-based model, in which the consultant and a trainee saw all new patients referred to the hospital, was gradually replaced with a team approach, based on continuous reconfiguration of the roles of the orthopaedic surgeon and rheumatologist and extending the roles of nurses, physiotherapists and podiatrists. This was achieved by: (i) protocol-based daily triage for all referrals to the most appropriate health professional in the team, by the senior out-patient nursing staff; (ii) allocation of appointments based on clinical priority, with a fast-track for urgent cases; and (iii) improvement of inter disciplinary communication, facilitating the retraction as well as the extension of traditional roles. RESULTS: Despite the number of GP referrals to the orthopaedic out-patient department at Stobhill nearly doubling in a period of 5 years, the out-patient waiting time decreased by about 50% (90 days from 182 days). This reduction in waiting times improved patient and GP satisfaction levels. We also noticed an improved morale and personal development of the health professionals as they saw patients appropriate to their skills and expertise. CONCLUSION: The team's experience demonstrates the effectiveness of a team approach in tackling what is often seen as the insoluble problem of orthopaedic waiting times. This is based on excellent communication and collaboration, with a clear aim of improving patient care that is evidence based. PMID- 15901378 TI - An algorithm to standardise the investigation of the undiagnosed traumatic painful adult hip: results at one year. AB - INTRODUCTION: A significant number of adults present to accident and emergency departments with a painful hip following a fall. When plain radiography is non diagnostic, it has been traditionally difficult to decide on further investigations as rapid access MRI is still unavailable in many NHS hospitals and, therefore, alternative methods of reliable investigation are required. PATIENTS & METHODS: An algorithm was designed for the management of these patients without the availability of MRI investigation. Over a 60-week period, 278 patients were admitted of whom 31 were adult patients with trauma-related hip pain with no fracture on plain radiography. RESULTS: We revealed 42% had fractures of the hip or pelvic girdle. None of the hip fractures deteriorated to a worse prognostic grade during the investigation process and no hip fractures were missed. CONCLUSIONS: This approach towards a challenging diagnostic problem has been successful in identifying all hip fractures, and no fracture has deteriorated to a worse prognostic group. PMID- 15901379 TI - Short-listing for orthopaedic specialist registrar posts--what is important? AB - INTRODUCTION: Competition for specialist registrar (SpR) posts in orthopaedics is high. The aim of the current study was to provide evidence-based advice for applicants applying for SpR posts in orthopaedics. METHODS: The short-listing forms of 273 applicants for orthopaedic SpR posts in South Thames (West) were reviewed. The experience of short-listed candidates was compared with those that were not. RESULTS: We have shown a high chance of being short-listed between 5 and 6 years after qualification with a sharp fall off either side of this. It is clear that a wide range of appropriate experience is more important than high volume and low quality experience in orthopaedics alone. CONCLUSION: Candidates who plan ahead and gain a broad experience have a better chance of progressing in an orthopaedic career. Good, early career advice is essential to achieve this. PMID- 15901380 TI - The use of skull X-rays in head injury in the emergency department--a changing practice. AB - INTRODUCTION: This is a consequential study of a previous audit, evaluating the role of skull X-rays in the emergency department in patients with head injuries, to see if the changes recommended, (implementation of The Royal College of Surgeons of England guidelines) have been adopted successfully and had the desired result. PATIENTS & METHODS: All patients who attended the accident and emergency department at the Royal Berkshire Hospital from 1-30 November 2003 with a diagnosis of head injury had their notes analysed for indications for skull X ray, presence of fracture and outcome. RESULTS: 278 patients were identified as having a head injury and had notes available, 19% (54/278) of these patients had a skull X-ray, of whom 31% (17/54) had a clearly documented indication. This shows a marked improvement from the previous audit when 50% (193/385) had a skull X-ray with only 7% (14/193) having a clearly documented indication. DISCUSSION: Following the introduction of new guidelines, a clear improvement in the practice of evidence-based medicine has been achieved; however, there is still room for further improvement and on-going education of staff and auditing of performance will help to ensure this continues. PMID- 15901381 TI - Appendix mass: do we know how to treat it? AB - INTRODUCTION: The traditional management of appendiceal mass has been an initial conservative approach followed by interval appendicectomy. More recently, the necessity of interval appendicectomy has been questioned by a growing amount of evidence in the surgical literature. The aim of this study was to review the available scientific evidence and to determine how appendiceal masses are currently being managed in the Mid-Trent region by general surgeons. PATIENTS & METHODS: A literature search using Medline, Embase, Cinahl, HMIC and Biosis was carried out. A personal or telephonic survey of all consultants and specialist registrars working in general surgery in the Mid-Trent region (n = 67) was conducted recording their management protocol of 3 different clinical scenarios- a 14-year-old boy, a 29-year-old female and a 68-year-old male. Responses of the questionnaire were entered to a database in Microsoft Access 2000 and analysed. RESULTS: The results showed that there was difference of opinion on the management of appendix mass in either scenario. Appendectomy (interval or emergency) is still practised by 75% of general surgeons in the Mid-Trent region and less that 25% manage asymptomatic appendix mass without interval appendectomy. Additionally, specialist registrars appear more likely not to offer patients interval appendicectomy after successful conservative management (P < 0.05). CONCLUSIONS: At present, there is no agreed consensus on the management of appendiceal mass. There is a need to develop a protocol for the management of this common problem. PMID- 15901382 TI - Do specialist registrars fail smokers? AB - INTRODUCTION: Medical and surgical SpRs were interviewed to determine what efforts they make to stop patients smoking. METHODS: Telephone interview of 53 SpRs in the Oxford region. RESULTS: The majority regularly asked whether patients smoked, few did anything about it. Surgical SpRs were less likely than medical SpRs to advise patients to stop, discuss smoking-related health problems, discuss the benefits of quitting or advise nicotine replacement therapy. Few SpRs had been trained to counsel smokers and less than half felt that their input helped patients to stop. CONCLUSION: Medical education is lacking in teaching students and junior doctors how to help patients stop smoking. PMID- 15901383 TI - Maximising SHO training by inclusion of research fellows into a novel hybrid rota. AB - INTRODUCTION: As a result of current government legislation, junior surgical trainees are increasingly working in shift patterns. Night shifts provide few training opportunities and recent debate has focused on the most effective way of providing appropriate surgical training in the face of these working pattern restrictions. METHODS: At York Hospital, we have recently implemented a new hybrid rota in general surgery which uses research fellows to cover the majority of night-time shifts at SHO level. RESULTS & DISCUSSION: Surgical trainees have benefited by spending a much greater proportion of their time working with patients during the day where training is more continuous, better supervised and more uniform. Research fellows have benefited in being able to maintain their clinical skills whilst having sufficient free time to pursue research during the day. Extending the role of the research fellow at night to include registrar duties could also release higher surgical trainees from some night-time service commitments and allow them to benefit from more day-time training. PMID- 15901384 TI - Implementation of interventional procedures guidance. PMID- 15901386 TI - Can performance as an undergraduate assist entry selection into surgical training programmes? PMID- 15901388 TI - The generality of negative hierarchically restrictive behaviours. AB - Previous research has shown that when an actor engages in a negative hierarchically restrictive behaviour, a strong correspondent trait attribution is made and this behaviour is expected to generalize across situations (Trafimow, 2001). This paper discusses three experiments that examined the effects of extreme situations and perceived morality of the actor on the way in which participants make trait attributions, and the extent to which those behaviours are expected to generalize to other situations. Findings from Experiments 1 and 2 indicate that even negative hierarchically restrictive behaviours do not always lead to strong correspondent inferences if the situation in which the initial behaviour was performed was sufficiently extreme. Experiment 3 served to support these findings and cleared up questions from the first two experiments. Findings supported the hypothesis that some situations inhibit negative trait attributions and behaviour generalizations. Furthermore, findings indicate that the perception of the morality of the actor determines whether or not a negative hierarchically restrictive behaviour will lead to a negative trait attribution and generalization. PMID- 15901389 TI - Attitude change and the public-private attitude distinction. AB - In this paper, we highlight the importance of the distinction between public and private attitudes in research on attitude change. First, we clarify the definitions of public and private attitudes by locating the researcher as a potential source of influence. In a test of this definition, we compare participant reports of potentially embarrassing behaviour and the study's importance between participants responding when a researcher has potential access to their reports (public condition), and participants whose reports the researcher has no potential access to (private condition). Participants high in public self-focus or low in defensive self-presentation reported the study to be more important in the public condition than the private condition. Further, participants in the public condition reported less frequency of engaging in embarrassing behaviours than those in the private condition, an effect not moderated by individual differences. We conclude that the public-private distinction is an essential element in attitude change theory. PMID- 15901390 TI - Predictive models of implicit and explicit attitudes. AB - Explicit attitudes have long been assumed to be central factors influencing behaviour. A recent stream of studies has shown that implicit attitudes, typically measured with the Implicit Association Test (IAT), can also predict a significant range of behaviours. This contribution is focused on testing different predictive models of implicit and explicit attitudes. In particular, three main models can be derived from the literature: (a) additive (the two types of attitudes explain different portion of variance in the criterion), (b) double dissociation (implicit attitudes predict spontaneous whereas explicit attitudes predict deliberative behaviour), and (c) multiplicative (implicit and explicit attitudes interact in influencing behaviour). This paper reports two studies testing these models. The first study (N = 48) is about smoking behaviour, whereas the second study (N = 109) is about preferences for snacks versus fruit. In the first study, the multiplicative model is supported, whereas the double dissociation model is supported in the second study. The results are discussed in light of the importance of focusing on different patterns of prediction when investigating the directive influence of implicit and explicit attitudes on behaviours. PMID- 15901391 TI - The goal-dependent automaticity of drinking habits. AB - In recent treatments of habitual social behaviour, habits are conceptualized as a form of goal-directed automatic behaviour that are mentally represented as goal action links. Three experiments tested this conceptualization in the context of students' drinking (alcohol consumption)habits. Participants were randomly assigned to conditions where either a goal related to drinking behaviour (socializing) was activated, or an unrelated goal was activated. In addition, participants' drinking habits were measured. The dependent variable in Experiments 1 and 2 was readiness to drink, operationalized by speed of responding to the action concept 'drinking' in a verb verification task. Experiment 3 used the uptake of a voucher to measure drinking behaviour. Findings supported the view that when habits are established, simply activating a goal related to the focal behaviour automatically elicits that behaviour. These findings are consistent with a goal-dependent conception of habit. Possibilities for interventions designed to attenuate undesirable habitual behaviours are considered. PMID- 15901392 TI - The effect of anxiety on impression formation: affect-congruent or stereotypic biases? AB - Two classes of theories propose that anxious individuals will form either more affect-congruent or more stereotypic impressions of others. These theories' predictions are not mutually exclusive. Eighty-one participants were examined to determine if either class of theories was more descriptive of the effect of anxiety on impression formation or whether a theory combining elements of both was more appropriate. Anxious participants read behavioural descriptions about an Australian Aboriginal target that were stereotypic, non-stereotypic, threatening, and non-threatening, and rated the target on traits that corresponded to the behavioural descriptions. Anxious participants formed impressions that were more affect-congruent, but not more stereotypic, than those formed by control participants. This result was replicated in a field study with 61 participants who were waiting to see a dentist. Future studies should examine the cognitive mechanisms that influence and underlie anxious affect-congruent impression formation. PMID- 15901393 TI - The subjective experience of partnership love: a Q Methodological study. AB - The social scientific literature seems increasingly to accept that defining partnership or 'romantic' love in a singular and timeless fashion might not be a realisable task (Fehr, 1988). One clearly discernible response to this situation takes the form of a turn towards studying the subjective experience of partnership love. The work of Marston, Hecht, Manke, McDaniel, and Reeder (1998) and Sternberg (1995, 1996, 1998) are highlighted as theoretical and methodological exemplars in this regard. Each successfully abstracts a range of cultural conceptions of partnership love from the subjective experiences of their participants. It is argued, however, that the subjective experiences of the participants themselves, and particularly the holistic or Gestalt nature of those experiences, are left behind in the process of abstraction. This new research seeks to rectify that situation. An alternative (but nonetheless related) approach to the study of partnership love is duly outlined and an illustrative Q Methodological study is reported which reveals eight distinct subjective experiences of partnership love at work amongst our participants (N = 50). Each of these holistic experiences is presented in the form of a narrative account. These accounts are then discussed and situated in relation to existing academic, cultural and historical conceptions of partnership love. PMID- 15901394 TI - Upward social comparison and self-concept: inspiration and inferiority among art students in an advanced programme. AB - We examined the role of social comparison in changes in the artistic self-concept of adolescents attending an advanced arts programme. Unfavourable comparisons that promoted a sense of inferiority and favourable comparisons that promoted inspiration were measured just prior to, in the first week of, and at the end of the 6 week programme. Consistent with the 'big fish little pond effect', inferiority comparisons made during the programme were associated with negative changes in self-concept. Consistent with the social comparison literature, however, inspiration comparisons made during the programme were associated with positive changes in self-concept. Rather than suggesting that exposure to highly talented peers is necessarily unfavourable, results suggest that the interpretation of the comparisons made in situ determines the favourability of such exposure. PMID- 15901395 TI - Giver-receiver asymmetries in gift preferences. AB - When people are asked to choose between gift items, givers and receivers sometimes show different patterns of preferences. The article reports four experimental studies exploring these giver-receiver asymmetries. Whereas givers tend to prefer exclusive, but smaller gift items, receivers appear to prefer less luxurious, and more useful gifts (Experiment 1). Givers prefer gift vouchers to cash, and are concerned about timing, whereas more receivers accept cash gifts, and claim that it does not matter if the gift arrives late (Experiment 2). One interpretation of the results could be that givers conform more strongly to gift conventions (cultural norms for gift exchange). It is further argued that these differences are not due to a perceived difference between self and others (Experiment 3), but rather because of situational differences. When receivers perform separate evaluations of gifts rather than joint evaluations (pairwise comparisons), they tend to change their preference pattern towards a higher rating of exclusive gift items (Experiment 4). PMID- 15901399 TI - A unified approach to building and controlling spiking attractor networks. AB - Extending work in Eliasmith and Anderson (2003), we employ a general framework to construct biologically plausible simulations of the three classes of attractor networks relevant for biological systems: static (point, line, ring, and plane) attractors, cyclic attractors, and chaotic attractors. We discuss these attractors in the context of the neural systems that they have been posited to help explain: eye control, working memory, and head direction; locomotion (specifically swimming); and olfaction, respectively. We then demonstrate how to introduce control into these models. The addition of control shows how attractor networks can be used as subsystems in larger neural systems, demonstrates how a much larger class of networks can be related to attractor networks, and makes it clear how attractor networks can be exploited for various information processing tasks in neurobiological systems. PMID- 15901400 TI - Synchronized firings in the networks of class 1 excitable neurons with excitatory and inhibitory connections and their dependences on the forms of interactions. AB - Synchronized firings in the networks of class 1 excitable neurons with excitatory and inhibitory connections are investigated, and their dependences on the forms of interactions are analyzed. As the forms of interactions, we treat the double exponential coupling and the interactions derived from it: pulse coupling, exponential coupling, and alpha coupling. It is found that the bifurcation structure of the networks depends mainly on the decay time of the synaptic interaction and the effect of the rise time is smaller than that of the decay time. PMID- 15901401 TI - A hierarchy of associations in hippocampo-cortical systems: cognitive maps and navigation strategies. AB - In this letter we describe a hippocampo-cortical model of spatial processing and navigation based on a cascade of increasingly complex associative processes that are also relevant for other hippocampal functions such as episodic memory. Associative learning of different types and the related pattern encoding recognition take place at three successive levels: (1) an object location level, which computes the landmarks from merged multimodal sensory inputs in the parahippocampal cortices; (2) a subject location level, which computes place fields by combination of local views and movement-related information in the entorhinal cortex; and (3) a spatiotemporal level, which computes place transitions from contiguous place fields in the CA3-CA1 region, which form building blocks for learning temporospatial sequences. At the cell population level, superficial entorhinal place cells encode spatial, context-independent maps as landscapes of activity; populations of transition cells in the CA3-CA1 region encode context-dependent maps as sequences of transitions, which form graphs in prefrontal-parietal cortices. The model was tested on a robot moving in a real environment; these tests produced results that could help to interpret biological data. Two different goal-oriented navigation strategies were displayed depending on the type of map used by the system. Thanks to its multilevel, multimodal integration and behavioral implementation, the model suggests functional interpretations for largely unaccounted structural differences between hippocampo-cortical systems. Further, spatiotemporal information, a common denominator shared by several brain structures, could serve as a cognitive processing frame and a functional link, for example, during spatial navigation and episodic memory, as suggested by the applications of the model to other domains, temporal sequence learning and imitation in particular. PMID- 15901404 TI - Theory of the snowflake plot and its relations to higher-order analysis methods. AB - The snowflake plot is a scatter plot that displays relative timings of three neurons. It has had rather limited use since its introduction by Perkel, Gerstein, Smith, and Tatton (1975), in part because its triangular coordinates are unfamiliar and its theoretical properties are not well studied. In this letter, we study certain quantitative properties of this plot: we use projections to relate the snowflake plot to the cross-correlation histogram and the spike triggered joint histogram, study the sampling properties of the plot for the null case of independent spike trains, study a simulation of a coincidence detector, and describe the extension of this plot to more than three neurons. PMID- 15901405 TI - Asymptotic theory of information-theoretic experimental design. AB - We discuss an idea for collecting data in a relatively efficient manner. Our point of view is Bayesian and information-theoretic: on any given trial, we want to adaptively choose the input in such a way that the mutual information between the (unknown) state of the system and the (stochastic) output is maximal, given any prior information (including data collected on any previous trials). We prove a theorem that quantifies the effectiveness of this strategy and give a few illustrative examples comparing the performance of this adaptive technique to that of the more usual nonadaptive experimental design. In particular, we calculate the asymptotic efficiency of the information-maximization strategy and demonstrate that this method is in a well-defined sense never less efficient--and is generically more efficient--than the nonadaptive strategy. For example, we are able to explicitly calculate the asymptotic relative efficiency of the staircase method widely employed in psychophysics research and to demonstrate the dependence of this efficiency on the form of the psychometric function underlying the output responses. PMID- 15901406 TI - Maximum likelihood set for estimating a probability mass function. AB - We propose a new method for estimating the probability mass function (pmf) of a discrete and finite random variable from a small sample. We focus on the observed counts--the number of times each value appears in the sample--and define the maximum likelihood set (MLS) as the set of pmfs that put more mass on the observed counts than on any other set of counts possible for the same sample size. We characterize the MLS in detail in this article. We show that the MLS is a diamond-shaped subset of the probability simplex [0,1]k bounded by at most k x (k-1) hyper-planes, where k is the number of possible values of the random variable. The MLS always contains the empirical distribution, as well as a family of Bayesian estimators based on a Dirichlet prior, particularly the well-known Laplace estimator. We propose to select from the MLS the pmf that is closest to a fixed pmf that encodes prior knowledge. When using Kullback-Leibler distance for this selection, the optimization problem comprises finding the minimum of a convex function over a domain defined by linear inequalities, for which standard numerical procedures are available. We apply this estimate to language modeling using Zipf's law to encode prior knowledge and show that this method permits obtaining state-of-the-art results while being conceptually simpler than most competing methods. PMID- 15901407 TI - Estimating entropy rates with Bayesian confidence intervals. AB - The entropy rate quantifies the amount of uncertainty or disorder produced by any dynamical system. In a spiking neuron, this uncertainty translates into the amount of information potentially encoded and thus the subject of intense theoretical and experimental investigation. Estimating this quantity in observed, experimental data is difficult and requires a judicious selection of probabilistic models, balancing between two opposing biases. We use a model weighting principle originally developed for lossless data compression, following the minimum description length principle. This weighting yields a direct estimator of the entropy rate, which, compared to existing methods, exhibits significantly less bias and converges faster in simulation. With Monte Carlo techinques, we estimate a Bayesian confidence interval for the entropy rate. In related work, we apply these ideas to estimate the information rates between sensory stimuli and neural responses in experimental data (Shlens, Kennel, Abarbanel, & Chichilnisky, in preparation). PMID- 15901408 TI - Spike timing precision and neural error correction: local behavior. AB - The effects of spike timing precision and dynamical behavior on error correction in spiking neurons were investigated. Stationary discharges-phase locked, quasiperiodic, or chaotic-were induced in a simulated neuron by presenting pacemaker presynaptic spike trains across a model of a prototypical inhibitory synapse. Reduced timing precision was modeled by jittering presynaptic spike times. Aftereffects of errors-in this communication, missed presynaptic spikes were determined by comparing postsynaptic spike times between simulations identical except for the presence or absence of errors. Results show that the effects of an error vary greatly depending on the ongoing dynamical behavior. In the case of phase lockings, a high degree of presynaptic spike timing precision can provide significantly faster error recovery. For nonlocked behaviors, isolated missed spikes can have little or no discernible aftereffects (or even serve to paradoxically reduce uncertainty in postsynaptic spike timing), regardless of presynaptic imprecision. This suggests two possible categories of error correction: high-precision locking with rapid recovery and low-precision nonlocked with error immunity. PMID- 15901409 TI - A new approach to spatial covariance modeling of functional brain imaging data: ordinal trend analysis. AB - In neuroimaging studies of human cognitive abilities, brain activation patterns that include regions that are strongly interactive in response to experimental task demands are of particular interest. Among the existing network analyses, partial least squares (PLS; McIntosh, 1999; McIntosh, Bookstein, Haxby, & Grady, 1996) has been highly successful, particularly in identifying group differences in regional functional connectivity, including differences as diverse as those associated with states of awareness and normal aging. However, we address the need for a within-group model that identifies patterns of regional functional connectivity that exhibit sustained activity across graduated changes in task parameters. For example, predictions of sustained connectivity are commonplace in studies of cognition that involve a series of tasks over which task difficulty increases (Baddeley, 2003). We designed ordinal trend analysis (OrT) to identify activation patterns that increase monotonically in their expression as the experimental task parameter increases, while the correlative relationships between brain regions remain constant. Of specific interest are patterns that express positive ordinal trends on a subject-by-subject basis. A unique feature of OrT is that it recovers information about functional connectivity based solely on experimental design variables. In particular, there is no requirement by OrT to provide either a quantitative model of the uncertain relationship between functional brain circuitry and subject variables (e.g., task performance and IQ) or partial information about the regions that are functionally connected. In this letter, we provide a step-by-step recipe of the computations performed in the new OrT analysis, including a description of the inferential statistical methods applied. Second, we describe applications of OrT to an event-related fMRI study of verbal working memory and H(2)15O-PET study of visuo-motor learning. In sum, OrT has potential applications to not only studies of young adults and their cognitive abilities, but also studies of normal aging and neurological and psychiatric disease. PMID- 15901410 TI - Investigating the fault tolerance of neural networks. AB - Particular levels of partial fault tolerance (PFT) in feedforward artificial neural networks of a given size can be obtained by redundancy (replicating a smaller normally trained network), by design (training specifically to increase PFT), and by a combination of the two (replicating a smaller PFT-trained network). This letter investigates the method of achieving the highest PFT per network size (total number of units and connections) for classification problems. It concludes that for non-toy problems, there exists a normally trained network of optimal size that produces the smallest fully fault-tolerant network when replicated. In addition, it shows that for particular network sizes, the best level of PFT is achieved by training a network of that size for fault tolerance. The results and discussion demonstrate how the outcome depends on the levels of saturation of the network nodes when classifying data points. With simple training tasks, where the complexity of the problem and the size of the network are well within the ability of the training method, the hidden-layer nodes operate close to their saturation points, and classification is clean. Under such circumstances, replicating the smallest normally trained correct network yields the highest PFT for any given network size. For hard training tasks (difficult classification problems or network sizes close to the minimum), normal training obtains networks that do not operate close to their saturation points, and outputs are not as close to their targets. In this case, training a larger network for fault tolerance yields better PFT than replicating a smaller, normally trained network. However, since fault-tolerant training on its own produces networks that operate closer to their linear areas than normal training, replicating normally trained networks ultimately leads to better PFT than replicating fault-tolerant networks of the same initial size. PMID- 15901411 TI - Novel chemiluminescence-inducing cocktails, part I: the role in light emission of combinations of luminal with SIN-1, selenite, albumin, glucose oxidase and Co2+. AB - It is known that many agents influence the capacity of cells to produce reactive oxygen species. However, assaying these agents, both those that stimulate and those that inhibit reactive oxygen production, can be complicated and time consuming. Here, a method is described in which two different cocktails are employed to stimulate luminol-dependent chemiluminescence (LDCL). These cocktails are comprised of luminol, with either sodium selenite [IV] (SEL) or tellurite [IV] (TEL) (where IV and VI refer to the 4+ or 6+ oxidation state of selenium or tellurium salts, respectively), morpholinosidonimine (SIN-1), serum albumin and Co(2+), called the SIN-1a (with selenite) and SIN1b (with tellurite) cocktails, respectively; or luminol with glucose oxidase (GO), sodium selenite [IV] and Co(2+), called the GO cocktail. The cocktails functioned best in Hank's balanced salt solution (HBSS) containing 1% glucose at pH 7.4, incubated at approximately 22 degrees C. Within 30-60 s there was a burst of luminescence, which lasted for 7-10 min. In 100% ethanol, the SIN-1 cocktails also generated LDCL to 70% of that produced in HBSS. Neither selenite [VI], seleno-cystine, seleno-methionine, nor the selenium-containing drug, ebselen, could replace SEL. Moreover, the effects of the NO-donor, SIN-1, could not be replicated by the oxyradical generators, xanthine-xanthine oxidase or hypochlorous acid. Only low levels of luminescence were generated by combinations of the peroxyl radical generator, 2,2'-azobis-2 amidinopropane dihydrochloride (AAPH) with either SEL or TEL. It is suggested that light emission induced by the SIN1 cocktail results from the oxidation of SEL [IV] to the [VI] state, possibly due to the generation of mixtures of superoxide, peroxide, peroxynitrite and also of unidentified oxidant species, catalyzed by CoCo(2+). However, the involvement of hydroxyl radicals in LDCL could not be confirmed by use of either dimethyl thiourea or by electron spin resonance (ESR). LDCL induced by the two cocktails is strongly reduced by phosphates, EDTA, deferoxamine, CuCo(2+), MnCo(2+), as well as by the "classical" antioxidants superoxide dismutase (SOD), ascorbate, vitamin E, uric acid or thiols. It is suggested that these chemiluminescence cocktail systems can be used to determine the total anti-oxidant capacities of biological fluids and commercially available anti-oxidants. PMID- 15901412 TI - Novel chemiluminescence-inducing cocktails, part II: measurement of the anti oxidant capacity of vitamins, thiols, body fluids, alcoholic beverages and edible oils. AB - Using two luminescence-inducing cocktails, two distinct patterns of inhibition of light by different anti-oxidants have been identified, comprising Group A, in which a complete inhibition of light emission which is then followed by re emergence of light, forming apparent S-shaped curves or similar shapes. This light pattern is induced by the "classical" anti-oxidants, ascorbate, vitamin E, uric acid, thiols, deferoxamine, as well as by anti-oxidant agents present in plasma, saliva, urine and in extracts derived from black coffee, and Group B, in which a gradually emerging "mound"-shaped pattern of light was seen with extracts from the Tibetan plant mixture PADMA-28, elderberry (Sambucol), grape seeds, green and black teas, apple, parsimony, red wines, edible oils and SOD. While the results with the Group A agents point to the presence of probably a single, major, anti-oxidants relatively sensitive to oxidation, Group B agents probably include a mixture of anti-oxidants which are more resistant to oxidation. It was also shown that agents from Group B could protect agents from Group A against consumption by the oxidants generated by the cocktails. It is proposed that these simple to use cocktails which probably generate a multiplicity of oxidants mimicking those generated by activated phagocytes, can rapidly assess the total anti-oxidant capacities (TAOC) in body fluids derived from patients suffering of excessive oxidative stress. Also, this technique may be useful in determining the content of dietary anti-oxidants recommended as supplements to enhance the resistance against excessive oxidation of lipids. PMID- 15901413 TI - Pharmacological and haematological results of rat skin burn injury treatment with Cu(II)2(3,5-diisopropylsalicylate)4. AB - This research was performed to determine whether or not treatment of burn-injured rats with Cu(II)2(3,5-diisopropylsalicylate)4(Cu(II)2(3,5-DIPS)4) facilitated recovery from burn-injury. Four groups of adult male rats received a standard skin burn 1 h before an initial subcutaneous treatment which was continued daily for three days with either 0, 5, 10 or 20micromol Cu(II)2(3,5-DIPS)4/kg body mass. A fifth group was given no treatment. A sixth group served as a non-burn injured non-treated normal control group. At 3 h and on days 1, 2, 3, 7 and 14 post-burn-injury blood samples were obtained from rats in all groups for the determination of leukocyte, platelet and erythrocyte counts, clotting times, hemoglobin and hematocrit values. Total protein and middle mass peptides in plasma, as well as plasma lipid and erythrocyte membrane peroxidation products were determined on days 7 and 14. Burn wound healing and body mass were determined daily from day 0 to 6 with a notation of crust rejection by day 14. Treatment with Cu(II)2(3,5-DIPS)4 produced effects consistent with a facilitation of Cu-dependent immune-mediated physiological inflammatory responses to burn injury. It is concluded that treatment of burn injury with Cu(II)2(3,5-DIPS)4 supports Cu-dependent physiological responses involved in overcoming burn injury, which may have been further optimized by continued treatment beyond day 2, the last day of treatment. PMID- 15901414 TI - Comparison between C57Bl/6 and C57Bl/10 mycobacterial mouse pleurisy with respect to cellular migration and nitric oxide production. AB - Mycobacterium bovis-BCG (BCG) and Mycobacterium leprae (ML) have opposite inflammatory properties. Mycobacteria-induced pleurisy in C57Bl/6 and C57Bl/10 mice was evaluated to establish if their innate responses could be comparable, verifying cellular migration and nitrite production. Kinetic responses after ML or BCG intrathoracic injection were compared in those mice, sharing the H-2(b) MHC haplotype. BCG led to acute eosinophilia and late neutrophilia in both mice. In C57Bl/6 late pleurisy, monocytes and neutrophil recruitment was dose- and iNOS dependent, inhibited by methotrexate but not by indomethacin. Pleural macrophages released nitrites ex vivo after 7 days of BCG stimulus, without "priming" and blocked by the nitrite inhibitor L-N5-(1-iminoethyl)-ornithine (L-NIO). ML did not induce cellular migration or nitrite production, independent of the mouse strain, timing, or number of bacilli. Although these mycobacteria have high homology, there was no effect of ML on BCG-evoked secondary cellular recruitment. Both C57Black mice trigger similar onset of inflammatory responses to these mycobacteria, so far can alternatively be used in experimental studies. PMID- 15901415 TI - PADMA-28, a traditional Tibetan herbal preparation, blocks cellular responses to bFGF and IGF-I. AB - The growth factors basic fibroblast growth factor (bFGF) and insulin-like growth factor 1 (IGF-I) have been implicated in the pathophysiology of atherosclerosis and restenosis. The Tibetan herbal preparation PADMA-28 (a mixture of 22 plants which is used as an anti-atherosclerosis agent) was tested for its ability to inhibit the mitogenic activity of bFGF and IGF-I, growth factors involved in restenosis, atherosclerosis and tumour progression. DNA synthesis and proliferation of vascular smooth muscle cells, in response to serum bFGF, thrombin, or combinations thereof, were abrogated in the presence of microgram amounts of both the aqueous and organic, partially purified, extracts of PADMA 28. These fractions also inhibited IGF-I-mediated proliferation, migration and invasion of tumour cells responsive to IGF-I. The inhibition by PADMA 28 was reversible upon removal of the PADMA extracts, indicating that the effects were not related to cell toxicity. These and other properties (i.e., anti-oxidant activity) of PADMA-28 may be responsible for its beneficial effect as an anti atherosclerotic agent, suggesting that this herbal preparation may have potential applications in the prevention of intimal hyperplasia and arterial stenosis secondary to coronary angioplasty and bypass surgery, as well as in the prevention and treatment of other vascular diseases and tumour growth and metastasis. PMID- 15901416 TI - Interleukin-1beta inhibits paw oedema induced by local administration of latex of Calotropis procera extracts. AB - Interleukin-1beta (IL-1beta), a pro-inflammatory cytokine, has been reported to exhibit anti-inflammatory properties in the carrageenan-induced paw oedema model. In the present study, we have evaluated the anti-inflammatory activity of IL 1beta against inflammation induced by local administration of the methanol extract of dried latex of Calotropis procera (MeDL) and compared it with that against carrageenan. The anti-inflammatory activity of standard anti-inflammatory drugs, phenylbutazone (PBZ) and dexamethasone (DEX), was also evaluated against both inflammagens. Injection of an aqueous solution of dried latex and MeDL into the sub-plantar surface of the rat paw produced intense inflammation with a peak response occurring within 2 h, while the peak inflammatory response with carrageenan was obtained at 3 h. Subcutaneous injection of IL-1beta was found to be more effective against the inflammatory response elicited by carrageenan (70% inhibition) as compared to MeDL (50% inhibition) at 20microg/kg dose. On the other hand, PBZ effectively inhibited the inflammatory response elicited by both MeDL and carrageenan, while DEX was more effective against carrageenan. Thus, our study indicates that the difference in the anti-inflammatory effect of IL-1beta against latex of C. procera extract and carrageenan is due to the release of different mediators released by these inflammagens. PMID- 15901417 TI - Evaluation of the anti-inflammatory and analgesic properties of the extract of Russelia equisetiformis (Schlecht & Cham) Scrophulariacae. AB - A methanolic extract of Russelia equisetiformis whole plant was studied for anti inflammatory and analgesic activities in rats and mice using carrageenan-induced rat paw oedema, acetic-acid-induced writhing and tail-flick test. The extract, at 10, 20 and 40 mg/kg, significantly (P <0.05) inhibited carrageenan-induced oedema in rats. Abdominal constriction induced by acetic acid was also inhibited by the extract, within the same dose range. The extract at the same dose also prolonged the latency period in the tail-flick response test, which was reverted by naloxone. The results suggested that the extract possesses potential anti inflammatory and analgesic properties. PMID- 15901418 TI - The new GMS contract: impact and implications for managing the changes. AB - BACKGROUND: In February 2003, a new General Practitioner (GP) contract was agreed between the profession's leaders and the government, which was later accepted following a national ballot of GPs. However, the ballot simply required respondents to vote for or against the proposal; it did not provide any opportunity to identify which aspects of the new contract were more or less acceptable. Since the proposed changes were far reaching, the implications of implementing and managing these were considerable. Consequently, some information about how GPs viewed various components of the new contract would enable a more targeted and effective management strategy to be developed that would facilitate the introduction of all aspects of the contract. OBJECTIVES: To survey GPs working within the West Midlands region regarding their opinions on each of the key features of the new contract. METHOD: A postal survey of 360 GPs was undertaken, using a specially devised questionnaire. RESULTS: Four factors emerged as the most acceptable aspects of the contract: option to opt out of out of-hours work, flexibility in the services provided, prediction of future income levels and linking practice to performance targets. Least acceptable were: performance monitoring systems, the new financial formula for calculating income, greater patient involvement in service development and 24/48 hour access. With regard to potential outcomes of the contract, the most positive were considered to be increased proportion of salaried GPs, increased salaries, appropriate quality standards for care, earlier retirement; the factors least likely to be of potential benefit were: reduction in occupational stress, simplification of the regulatory framework, improved equity of workload and improved staff retention. Further analysis of the results using inferential statistics revealed a range of subgroup differences in reaction to the contract. CONCLUSION: Overall, those aspects of the new contract that are perceived to reduce workload and enhance salary were supported, while those that increase targets and bureaucracy were not. Generally, there was only moderate support for the changes, which could be explained by a general scepticism about any top-down modifications, the practicality and power of the changes to impact upon practice and/or a genuine belief that the modifications are unacceptable. Taken together, these results provide an indicative focus for managing the implementation of the new contract, especially with regard to its least acceptable components and the emerging differences between subgroups of GPs. PMID- 15901419 TI - Strategies for multi-hospital networks: a framework. AB - The last two decades have seen the significant emergence of multi-hospital networks (MHNs) in the health sectors of all industrialized countries. Such networks seem to represent an organizational choice, which provides interesting opportunities to cope with cost and quality issues stressing the health field. This work proposes and discusses a framework for strategies that can be implemented by MHNs to achieve projected benefits. The estimated advantages are then weighed against unexpected or underrated drawbacks and empirical evidence. The conclusions point out MHNs as viable future alternatives for freestanding hospitals looking to improve their efficiency and financial stability, on condition that a 'proper management' of their consolidation processes is carried out. This requires extensive pre-deal analysis to critically assess what benefits could be achieved through the network's formation, as well as post-deal perseverance in implementing consolidation strategies to their full. In this perspective, specific advices apt to minimize the risk of creating MHNs in the wake of a management fashion instead of through sound rational assessments are also discussed. PMID- 15901420 TI - From the trenches: views from decision-makers on health services priority setting. AB - Due to resource scarcity, health organizations worldwide must decide what services to fund and, conversely, what services not to fund. One approach to priority setting, which has been widely used in Britain, Australia, New Zealand and Canada, is programme budgeting and marginal analysis (PBMA). To date, such activity has primarily been based at a micro level, within programmes of care. In order to institute and refine the PBMA framework at a macro level across major service areas within a single health authority, researchers and decision-makers in Alberta embarked on a participatory action research project together. This paper identifies key issues of importance to decision-makers in a real-world priority-setting context. Themes discussed include making comparisons across disparate patient groups, dealing with political factors, using relevant forms of evidence, recognizing innovations and involving the public. The in-depth insight gained through this qualitative analysis will enable future refinement of PBMA at a macro level in the health authority under study, and should also serve to inform priority-setting activity in regionalized contexts elsewhere. In identifying aspects of priority setting that are important to decision-makers, researchers can also be better informed with respect to real-world processes. PMID- 15901421 TI - Using the balanced scorecard to characterize benefits of integration in the safety net. AB - The purpose of this study was to develop a comprehensive framework depicting the potential benefits of integration among health-care providers that serve vulnerable populations. Research teams interviewed participants in 12 integrated functions across seven community health-centre-led networks. Functions included clinical processes; managed care contracting; and administrative services such as human resources, finance, and information systems. Using a Balanced Scorecard framework, benefits were identified across financial, customer, internal business, and learning and growth perspectives. Financial benefits were more frequently cited relative to managed care and administrative functions than relative to clinical functions. Clinical functions were frequently characterized by perceived improvements in patient care quality, while managed-care functions appeared to yield most benefits in access. Administrative functions were most often associated with improvements in internal business operations. There were substantial findings in learning and growth across all three types of integration, in keeping with the early stages of the integrated functions in the study. Findings imply that integration among health-care providers yields a wide range of benefits, but not necessarily quickly or financial in nature. PMID- 15901422 TI - The estimation of distributions and the minimum relative entropy principle. AB - Estimation of Distribution Algorithms (EDA) have been proposed as an extension of genetic algorithms. In this paper we explain the relationship of EDA to algorithms developed in statistics, artificial intelligence, and statistical physics. The major design issues are discussed within a general interdisciplinary framework. It is shown that maximum entropy approximations play a crucial role. All proposed algorithms try to minimize the Kullback-Leibler divergence KLD between the unknown distribution p(x) and a class q(x) of approximations. However, the Kullback-Leibler divergence is not symmetric. Approximations which suppose that the function to be optimized is additively decomposed (ADF) minimize KLD(q||p), the methods which learn the approximate model from data minimize KLD(p||q). This minimization is identical to maximizing the log-likelihood. In the paper three classes of algorithms are discussed. FDA uses the ADF to compute an approximate factorization of the unknown distribution. The factors are marginal distributions, whose values are computed from samples. The second class is represented by the Bethe-Kikuchi approach which has recently been rediscovered in statistical physics. Here the values of the marginals are computed from a difficult constrained minimization problem. The third class learns the factorization from the data. We analyze our learning algorithm LFDA in detail. It is shown that learning is faced with two problems: first, to detect the important dependencies between the variables, and second, to create an acyclic Bayesian network of bounded clique size. PMID- 15901424 TI - Editorial introduction: special issue on estimation of distribution algorithms. PMID- 15901425 TI - Population-based continuous optimization, probabilistic modelling and mean shift. AB - Evolutionary algorithms perform optimization using a population of sample solution points. An interesting development has been to view population-based optimization as the process of evolving an explicit, probabilistic model of the search space. This paper investigates a formal basis for continuous, population based optimization in terms of a stochastic gradient descent on the Kullback Leibler divergence between the model probability density and the objective function, represented as an unknown density of assumed form. This leads to an update rule that is related and compared with previous theoretical work, a continuous version of the population-based incremental learning algorithm, and the generalized mean shift clustering framework. Experimental results are presented that demonstrate the dynamics of the new algorithm on a set of simple test problems. PMID- 15901426 TI - Globally multimodal problem optimization via an estimation of distribution algorithm based on unsupervised learning of Bayesian networks. AB - Many optimization problems are what can be called globally multimodal, i.e., they present several global optima. Unfortunately, this is a major source of difficulties for most estimation of distribution algorithms, making their effectiveness and efficiency degrade, due to genetic drift. With the aim of overcoming these drawbacks for discrete globally multimodal problem optimization, this paper introduces and evaluates a new estimation of distribution algorithm based on unsupervised learning of Bayesian networks. We report the satisfactory results of our experiments with symmetrical binary optimization problems. PMID- 15901427 TI - Estimation of distribution algorithms with Kikuchi approximations. AB - The question of finding feasible ways for estimating probability distributions is one of the main challenges for Estimation of Distribution Algorithms (EDAs). To estimate the distribution of the selected solutions, EDAs use factorizations constructed according to graphical models. The class of factorizations that can be obtained from these probability models is highly constrained. Expanding the class of factorizations that could be employed for probability approximation is a necessary step for the conception of more robust EDAs. In this paper we introduce a method for learning a more general class of probability factorizations. The method combines a reformulation of a probability approximation procedure known in statistical physics as the Kikuchi approximation of energy, with a novel approach for finding graph decompositions. We present the Markov Network Estimation of Distribution Algorithm (MN-EDA), an EDA that uses Kikuchi approximations to estimate the distribution, and Gibbs Sampling (GS) to generate new points. A systematic empirical evaluation of MN-EDA is done in comparison with different Bayesian network based EDAs. From our experiments we conclude that the algorithm can outperform other EDAs that use traditional methods of probability approximation in the optimization of functions with strong interactions among their variables. PMID- 15901428 TI - Drift and scaling in estimation of distribution algorithms. AB - This paper considers a phenomenon in Estimation of Distribution Algorithms (EDA) analogous to drift in population genetic dynamics. Finite population sampling in selection results in fluctuations which get reinforced when the probability model is updated. As a consequence, any probability model which can generate only a single set of values with probability 1 can be an attractive fixed point of the algorithm. To avoid this, parameters of the algorithm must scale with the system size in strongly problem-dependent ways, or the algorithm must be modified. This phenomenon is shown to hold for general EDAs as a consequence of the lack of ergodicity and irreducibility of the Markov chain on the state of probability models. It is illustrated in the case of UMDA, in which it is shown that the global optimum is only found if the population size is sufficiently large. For the needle-in-a haystack problem, the population size must scale as the square root of the size of the search space. For the one-max problem, the population size must scale as the square-root of the problem size. PMID- 15901429 TI - Space complexity of estimation of distribution algorithms. AB - In this paper, we investigate the space complexity of the Estimation of Distribution Algorithms (EDAs), a class of sampling-based variants of the genetic algorithm. By analyzing the nature of EDAs, we identify criteria that characterize the space complexity of two typical implementation schemes of EDAs, the factorized distribution algorithm and Bayesian network-based algorithms. Using random additive functions as the prototype, we prove that the space complexity of the factorized distribution algorithm and Bayesian network-based algorithms is exponential in the problem size even if the optimization problem has a very sparse interaction structure. PMID- 15901430 TI - How do we move forward? PMID- 15901431 TI - Patient-active prevention in primary dental care: a characterisation of general practices in Northern Ireland. AB - AIM: To investigate the preventive orientation of general dental practices by examining their patient-active prevention activities, practice policies for prevention and employment strategies. METHOD: All general dental practices located within the region of the Eastern Health and Social Services Board (EHSSB) in Northern Ireland were contacted and invited to participate. A questionnaire assessed practice characteristics, patient-active prevention, practice policies and employment strategies. The principal general dental practitioner (GDP) was invited to complete the questionnaire. RESULTS: Seventy-seven per cent of practices participated. Nearly all the practices provided patient-active prevention; however, lower proportions of dentists provided advice on diet for dental caries (3%), oral hygiene instruction, fluoride toothpaste use and interdental cleaning (7%) with each new course of dental treatment. 'Patient active' prevention practices were 5.8 times more likely to employ a hygienist and 5.3 times more likely to have a practice policy to screen for oral cancer. CONCLUSIONS: These findings suggest that the employment of a hygienist is central to the reorientation of primary dental care. The Government must be encouraged to provide the financial means to allow primary care to shift from being disease centred to health-focused. PMID- 15901432 TI - The use of ozone in dentistry and medicine. AB - There is growing interest in the use of ozone in oral healthcare and the National Institute for Clinical Excellence (NICE) is at present reviewing the evidence for its effectiveness in the management of occlusal and plain surface caries. These are only two of the clinical problems for which ozone can, and has, been used; it has also been employed for a wide variety of other purposes in both dentistry and medicine. This pale blue-coloured gas plays an important role as a natural constituent in the higher layer of the Earth's atmosphere. There is growing evidence that it can be employed as a useful therapeutic agent. This paper reviews its therapeutic uses to date and suggests its possible future clinical applications. Consumer demands for this strong oxidant may increase as the general public becomes increasingly aware of its therapeutic capacity and the non invasive manner in which it can be administered. PMID- 15901433 TI - Evaluation of the potential risk of occupational asthma in dentists exposed to contaminated dental unit waterlines. AB - INTRODUCTION: Most of the organisms isolated from dental unit waterlines (DUWL) are Gram-negative bacteria, which contain cell wall endotoxin. A consequence of endotoxin exposure is the exacerbation of asthma. OBJECTIVES: This study examined the prevalence and onset of asthma among dentists and determined whether or not these were associated with the microbiological quality of DUWL in their practices. METHODS: 266 randomly selected dentists (100 from rural Northern Ireland, 166 from London) completed a health questionnaire, which included questions on prevalence and time of onset of asthma. Water samples taken from the dental handpieces and surgery washbasin cold taps in all the practices were analysed using standard techniques. The questionnaire data were evaluated using both single and multivariable logistic regression. The variables considered were: smoking; surgery location; time treating patients per week; DUWL counts of Pseudomonas aeruginosa, total Pseudomonas spp., fungi, Mycobacterium spp., total aerobic colony counts (ACC) at 22 degrees C and 37 degrees C. RESULTS: There was no significant association between any of the variables tested in dentists and a history of asthma. A subgroup analysis was performed on dentists (n=33) who reported developing asthma since they started dental training. The final multivariable model indicated that passive smoking (OR 0.08, 95% CI 0.01-0.87, P=0.038) and total aerobic counts of >200 cfu/ml at 37 degrees C (OR 6.72, 95% CI 1.15-39.24, P=0.034) were significant variables for developing asthma since starting training as a dentist. ACC were significantly higher in London (P<0.0001) and London dentists were more likely to have developed asthma since they started training than their Northern Ireland counterparts (OR 4.4, 95% CI 1.09-17.72, P=0.033). CONCLUSIONS: This study suggests that the temporal onset of asthma may be associated with occupational exposure to contaminated DUWL among dentists in London and Northern Ireland. PMID- 15901434 TI - The effect of infection-control barriers on the light intensity of light-cure units and depth of cure of composite. AB - AIM: Dental curing lights are vulnerable to contamination with oral fluids during routine intra-oral use. This controlled study aimed to evaluate whether or not disposable transparent barriers placed over the light-guide tip would affect light output intensity or the subsequent depth of cure of a composite restoration. METHODS: The impact on light intensity emitted from high-, medium- and low-output light-cure units in the presence of two commercially available disposable infection-control barriers was evaluated against a no-barrier control. Power density measurements from the three intensity light-cure units were recorded with a radiometer, then converted to a digital image using an intra-oral camera and values determined using a commercial computer program. For each curing unit, the measurements were repeated on ten separate occasions with each barrier and the control. Depth of cure was evaluated using a scrape test in a natural tooth model. RESULTS: At each level of light output, the two disposable barriers produced a significant reduction in the mean power density readings compared to the no-barrier control (P<0.005). The cure sleeve inhibited light output to a greater extent than either the cling film or the control (P<0.005). Only composite restorations light-activated by the high level unit demonstrated a small but significant decrease in the depth of cure compared to the control (P<0.05). CONCLUSION: Placing disposable barriers over the light-guide tip reduced the light intensity from all three curing lights. There was no impact on depth of cure except for the high-output light, where a small decrease in cure depth was noted but this was not considered clinically significant. Disposable barriers can be recommended for use with light-cure lights. PMID- 15901436 TI - Transmigration of a maxillary canine. A case report. AB - In the oral cavity, transmigration is defined as a tooth that crosses the mid line by more than half its length. Following extensive literature review, it was found that, although well documented with respect to mandibular canines, there were only two publications detailing maxillary cuspid transmigration. This report presents a case of transmigration where the left maxillary canine gradually crossed the mid-line and migrated to the right side in a patient with hemifacial microsomia and cleft palate. The Mupparapu classification of the migratory pattern of the mandibular canines is discussed. Various clinical considerations, as well as guidelines for general practitioners to diagnose and manage such a condition, are also discussed. PMID- 15901437 TI - Digital imaging in pathology: the case for standardization. AB - The process of digital imaging in microscopy is a series of operations, each contributing to the quality of the final image that is displayed on the computer monitor. The operations include sample preparation and staining by histology, optical image formation by the microscope, digital image sampling by the camera sensor, postprocessing and compression, transmission across the network and display on the monitor. There is an extensive literature about digital imaging and each step of the process is fairly well understood. However, the complete process is very hard to standardize or even to understand fully. The important concepts for pathology imaging standards are: (1) systems should be able to share image files, (2) the standards should allow the transmission of information on baseline colours and recommended display parameters, (3) the images should be useful to the pathologist, not necessarily better or worse than direct examination of a slide under the microscope, (4) a mechanism to evaluate image quality objectively should be present, (5) a mechanism to adjust and correct the minor errors of tissue processing should be developed, (6) a public organization should support pathologists in the development of standards. PMID- 15901438 TI - Information on telemedicine. AB - Although the practice of telemedicine is still not considered part of mainstream health care, there has been a steady growth in information about telemedicine since 1999. The increasing body of literature about telemedicine, the staying power of various telemedicine societies and Websites, and the growing number of well-attended conferences about telemedicine all attest that interest in and the practice of telemedicine worldwide are very much alive and well. Searching for information has also become easier with the advent of better and faster search engines, particularly Google. Nonetheless, many Websites are out of date and the semantic confusion between 'telemedicine' and other terms such as 'telehealth' means that considerable perseverance is required to sift the wheat from the chaff. There is a wide variety of both electronic and print resources that have proven to be reliable sources of information. These include: the Internet (Web), books and reports, journals and trade magazines, telemedicine societies and conferences. While telemedicine cannot yet be considered to be part of mainstream health care, it has become a more familiar part of health terminology worldwide. PMID- 15901439 TI - Education through telemedicine networks: setting quality standards. AB - Quality standards for educational programming have received limited attention in telemedicine. We selected five sets of standards from the distance education literature established by: (1) the American Council on Education; (2) the American Distance Education Consortium; (3) the Council of Regional Accrediting Commissions; (4) the Distance Education and Training Council; (5) the Innovations in Distance Education Project. The standards were reviewed to determine the purposes they were intended to serve and the process by which they were established. The content of the five sets of standards were summarized around the 'four commonplaces' of education: learner, teacher, curriculum and context. Four major findings emerged. First, none of the sets of standards addresses all of the issues that are potentially relevant to telemedicine education; all emphasize certain topics while neglecting others. Second, there are some important aspects of telemedicine that are not addressed at all, such as patient confidentiality. Third, the standards generally provide a framework for defining high quality in distance education, leaving to those at the local level the task of deciding how a standard applies in their setting. Finally, the standards reviewed have many elements that could potentially apply to telemedicine education. Setting quality standards for education through telemedicine requires a systematic approach and a means for continuous improvement of those standards. PMID- 15901440 TI - A review of telemedicine in Uzbekistan. AB - The government of Uzbekistan has recognized the need for telemedicine. With the support of several international groups, three telemedicine programmes have been established. This international assistance has provided equipment and training. There is a national network of emergency medical services, and two e-referral projects, one led by the Swinfen Charitable Trust and the other by NATO. Although these have been successful, they continue to face similar difficulties. However, telemedicine has shown its value. PMID- 15901441 TI - Remote interpretation in medical encounters: a systematic review. AB - We conducted a systematic review of remote interpretation. Any published or unpublished research article that had telephone or videoconferenced interpretation as an intervention was included in the review. Nine articles were identified: seven on telephone and two on videoconferenced interpretation. Only one study was a randomized controlled trial. Remote interpretation was associated with shorter intervals between consultations, but there were no consistent differences in relation to consultation length. Client and doctor satisfaction was as good with remote interpretation as with face-to-face interpretation, but interpreters preferred face-to-face interpretation. Although the costs associated with remote interpreting are higher, especially in the case of videoconferencing, these may be offset by efficiency gains. The review suggests that remote interpretation is an acceptable and accurate alternative to traditional methods, although the associated costs are higher. PMID- 15901442 TI - Role of telehealth in seating clinics: a case study of learners' perspectives. AB - We conducted a qualitative case study of the introduction of telehealth in a seating clinic, which was an existing outreach service between two hospitals in Alberta, approximately 300 km apart. Interviews were conducted with the staff who were involved in planning and implementing the telehealth initiative. The study showed that, from the perspective of the staff (who were learners), implementation of telehealth in seating clinics differs from other less tactile telehealth applications in certain ways: (1) the importance of multidisciplinary teams in the procedures, (2) the importance of proper visualization and communication among the staff to convey the pressure changes and measurements to the technicians at the major centre to help them build or adjust the seating devices and (3) the reluctance of staff to trust others' judgements. Planning of service provision and telelearning for seating clinics requires the involvement of staff at all stages. Thus, the implementation of telehealth should be a stepwise process, allowing a highly interactive approach, without affecting the multidisciplinary nature of seating clinics. PMID- 15901443 TI - An observational study of veterans with diabetes receiving weekly or daily home telehealth monitoring. AB - We assessed the utilization of health-care services and clinical outcomes in veterans with diabetes who were enrolled in two care coordination/home telehealth programmes. One group of patients was monitored weekly (n = 197), with more intensive evaluations, while the other was monitored daily (n = 100), but less intensively. Although patients in the two groups were fairly similar in demographic terms and in their clinical characteristics at baseline, they had different service utilization patterns during the 12-month pre-enrollment period. Over the 12-month study period, the proportion of one or more hospital admissions and number of bed days of care decreased in the daily monitoring group, and increased in the weekly monitoring group, more or less doubling in the former and being halved in the latter. Unscheduled primary care clinic visits were lower in the daily monitored group than in the weekly monitored group. The differences between the two groups were significant (P < 0.01). There were no significant differences between the groups in the clinical outcomes. Future research should employ randomized controlled trial designs to determine if intensities of home monitoring lead to differences in service utilization and health outcomes. PMID- 15901444 TI - Remote psychotherapy for terminally ill cancer patients. AB - We conducted a feasibility study of remote psychotherapy in 10 terminally ill cancer patients with diagnoses of adjustment disorder or major depression. Subjects received six sessions of individual cognitive therapy with the same therapist. Sessions alternated between face-to-face sessions and remote sessions delivered by analogue videophone. After each therapy session, a brief questionnaire was used to evaluate the subjects' level of satisfaction with the session, sense of connectedness to the therapist and overall progress being made in the therapy. Nine patients completed the study. Of 53 completed therapy sessions, 21 were by videophone and 32 were conducted face to face. Participants reported strong positive perceptions and acceptance after almost all therapy sessions, regardless of service delivery mode. The study suggests that there may be a role for the delivery of psychotherapy using low-bandwidth videophones. PMID- 15901446 TI - Recruiting gamete donors: response to Craft and Thornhill. AB - This response challenges the notion that paying more money, in whatever form, will overcome the anxieties of gamete donors regarding the removal of anonymity. An overview of the literature on payment and donor recruitment indicates that this is a complex issue. That literature does provide 'pointers' to the types of persons who should be 'targeted' in an information sharing and non-payment system. The experience in Sweden, both in relation to donor recruitment and parents' information-sharing with their children, is clarified, as the evidence is not always presented accurately. PMID- 15901447 TI - Further HFEA restrictions on egg donation in the UK: two strikes and you're out! AB - The UK and Europe have lagged far behind the USA in the number of egg donation cycles performed over the past two decades. This disparity has been largely attributed to governmental restraints placed on the method within these locales, combined with the lack of regulation in the USA. Severely limiting donor compensation and requiring donors to be identified 18 years or more after their participation will undoubtedly lead to the demise of egg donation as the UK now knows it. Throwing more money at the problem, in the form of increased donor compensation, is unlikely to fix the shortage of participants that is to come. As already witnessed, increasing numbers of women and couples will probably seek treatment outside their native borders to escape the unreasonable demands placed upon their personal liberties by government. Regulation will promote 'reproductive tourism' as patients seek care in areas of the world where self determination is protected by law. PMID- 15901448 TI - Too late for change, too early to judge, but an oxymoron will not solve the problem. AB - In the face of an impending change in the recruitment of gamete donors in the UK, many predict a decrease in the number of donations. It is argued that the dangers of inordinate compensation may only increase this possible dearth by deterring altruistic donors, and that large payments may be especially prejudicial to the interests and welfare of children born as a result of gamete donation. Thus, "an 'all-inclusive' financial allowance to ...donor(s)" appears to be a contradictory statement (an oxymoron) that is unlikely to resolve the practical problems arising in gamete donor recruitment in the UK. PMID- 15901449 TI - Belgian legislation and the effect of elective single embryo transfer on IVF outcome. AB - In order to reduce the number of multiple pregnancies following IVF, the Belgian government agreed to reimburse laboratory expenses for six IVF cycles up to the age of 42 years, in exchange for restriction of the number of embryos replaced. Data on assisted reproduction outcome before and after the introduction of this new legislation were analysed retrospectively in terms of implantation, pregnancy and multiple pregnancy rates. After the introduction of the new law, the percentage of single embryo transfer increased from 14 to 49%. Implantation rates were 25.9 and 23% respectively. There was no difference in the overall pregnancy rate before and after the introduction (36 versus 37%). Twin pregnancies, however, decreased from 19 to 3%. These findings indicate that elective single embryo transfer significantly decreases the twin pregnancy rate without a reduction in the overall pregnancy rate. PMID- 15901450 TI - Batch-to-batch consistency of human-derived gonadotrophin preparations compared with recombinant preparations. AB - Different gonadotrophin preparations derived from human urine or manufactured by recombinant technology are currently used in clinical practice for the treatment of infertility. It has been widely assumed that gonadotrophin products manufactured by recombinant technology have better batch-to-batch consistency compared with human-derived preparations and that this potentially will be shown to provide a more constant clinical response, but there is little evidence for either statement. This study compared the batch-to-batch consistency between urinary-derived and recombinant manufactured gonadotrophin preparations using standard analytical techniques, as well as a novel in-vitro follicle bioassay to evaluate the consistency of the biological response at the target organ. Oligosaccharide isoform profiling, immunoassay testing, size exclusion chromatography analysis and in-vitro bioassay testing of urinary derived gonadotrophin preparations (MENOPUR and BRAVELLE) confirm that these products display a high degree of batch-to-batch consistency, similar to recombinant FSH (GONAL-f) either filled by mass or bioassay. The data also suggest that the batch to-batch variation is independent of the manufacturing procedure (filled-by bioassay or filled-by-mass) for the recombinant preparation (Gonal-f), but that the total FSH bioactivity delivered from a single dose preparation after reconstitution differs between the two manufacturing procedures. PMID- 15901451 TI - Factors affecting outcome after ICSI with spermatozoa retrieved from cryopreserved testicular tissue in non-obstructive azoospermia. AB - There is a lack of data regarding variables affecting the treatment outcome for non-obstructive azoospermia when spermatozoa from cryopreserved testicular specimens are utilized for ICSI. The objective of the present retrospective analysis was to investigate the effect of various parameters on treatment outcome in such cases. One hundred and sixty-five couples with non-obstructive azoospermic males undergoing a total of 297 cycles were included. In all cases the testicular tissue retrieved by multiple open-biopsy testicular sperm extraction was stored in liquid nitrogen and, after thawing, only mature spermatozoa were used for ICSI. When no motile spermatozoa were recovered, immotile spermatozoa were used. In 159 cycles, motile spermatozoa were utilized for ICSI, while in 138 cycles immotile spermatozoa were utilized. Higher normal fertilization rate (60.4 +/- 3.1 versus 51.3 +/- 1.6%, P < 0.05), number of embryos transferred (2.8 +/- 0.06 versus 2.6 +/- 0.04, P < 0.05), modified cumulative embryo score (31.2 +/- 1.6 versus 23.9 +/- 0.8, P < 0.001), and proportion of motile spermatozoa injected (67.8 versus 49.8%, P < 0.05) were observed in cycles that resulted in clinical pregnancies. Binary logistic regression analysis showed that sperm motility (odds ratio 2.06, 95% CI 1.1-3.9, P < 0.05), but not woman's age, number of treatment cycle, type of GnRH-analogue used for pituitary suppression, number of oocytes retrieved or number of embryos transferred was a significant determinant of the likelihood of clinical pregnancy. In conclusion, sperm motility after freeze/thawing of testicular tissue is the major determinant of the success of ICSI in non-obstructive azoospermia. PMID- 15901452 TI - Cumulative pregnancy rate after ICSI with cryopreserved testicular tissue in non obstructive azoospermia. AB - The aim of the present study was to describe a simplified and inexpensive method of testicular tissue freezing, to assess the cumulative clinical pregnancy rate (CPR) by this technique, and to provide useful information for counselling couples with non-obstructive azoospermia. One hundred and sixty-five couples with non-obstructive azoospermic males pursuing assisted conception, from December 1995 to December 2002, were included. In all cases, the testicular tissue retrieved by open multiple-biopsy (both sides, by testicular sperm extraction) was frozen using a simple liquid nitrogen vapour freezing technique and was stored in liquid nitrogen thereafter. Only mature spermatozoa were used for intracytoplasmic sperm injection (ICSI) after thawing. Expected CPR were calculated using the Kaplan-Meier survival analysis. A total of 281 cycles were performed resulting in 53 clinical pregnancies. Crude and expected CPR (95% confidence intervals) after three cycles were 32.1 (25.7-40.1) and 55.7% (37.0 74.4) respectively. In conclusion, this simplified method for freezing testicular tissue resulted in a satisfactory outcome after ICSI in cases of non-obstructive azoospermia. PMID- 15901453 TI - Prevalence of chromosome defects in azoospermic and oligoastheno-teratozoospermic South Indian infertile men attending an infertility clinic. AB - The prevalence of chromosomal abnormalities in azoospermic and oligoastheno teratozoospermic infertile men of South Indian origin undergoing assisted reproductive technologies was evaluated. In addition, the study aimed to investigate new abnormal karyotypes involving autosomes in azoospermia and sex chromosomes in oligoastheno-teratozoospermic individuals that are supposed to be rare. Metaphase chromosomes of 744 infertile men, including 272 men with azoospermia and 472 men with oligoastheno-teratozoospermia (OAT), were analysed using Giemsa-trypsin-Giemsa banding and fluorescence in-situ hybridization (FISH) wherever necessary. Chromosomal abnormalities were observed in 59 (7.9%) individuals of the total studied population. Among these, 30 out of 272 (11.0%) azoospermic men and 29 out of 472 (6.1%) infertile men with OAT showed chromosomal abnormalities. A strong and statistically significant association (OR = 1.89; P = 0.0235) of chromosomal abnormalities and sex chromosome abnormalities (OR = 4.29; P = 0.001) with azoospermia when compared with OAT was observed. In addition, six autosomal abnormalities associated with azoospermia and two abnormalities involving Y chromosome, which include a novel karyotype (mos 46,XY/51,XYYYYYY) in OAT individuals, were detected. PMID- 15901454 TI - Effect of inner myometrium fibroid on reproductive outcome after IVF. AB - To evaluate the influence of inner myometrium fibroids (myomas) on the outcome of IVF cycles, a retrospective agematched controlled study was performed at SISMeR Reproductive Medicine Unit. The study group included 129 IVF/intracytoplasmic sperm injection cycles in 75 patients with one or more intramural and/or submucosal fibroids, while the control group consisted of 129 cycles in 127 patients without fibroids. The two groups were similar for mean oestradiol concentration at human chorionic gonadotrophin administration (1205.16 +/- 874 versus 1395 +/- 821 pg/ml), mean number of transferred embryos (2.02 +/- 0.4 versus 2.14 +/- 0.6) and clinical pregnancy rate (34.9 versus 41.1%). Conversely, the implantation rate was significantly lower in the study group (18.0%) than in the control group (26.5%; chi(2) = 4.81, P < 0.05), whereas the rate of spontaneous abortion demonstrated an opposite trend (40 versus 18.9%; chi(2) = 4.34, P < 0.05). Further research should be aimed at classifying fibroids on the basis of their location, especially when they are positioned in the junctional zone of the myometrium. Whether this classification will be superior in predicting the impact of fibroids on the reproductive outcome should be elaborated in a large multicentric study. PMID- 15901455 TI - Cell polarity during folliculogenesis and oogenesis. AB - Polarity is an important aspect of oogenesis and early development for many animal groups, but only recently it has become relevant to the study of mammals. Mammalian oocyte development occurs through tight coordination and interaction between all ovarian structures. In fact, bi-directional communication between the oocyte and its companion granulosa cells (GC) in the ovarian follicle seems essential for GC proliferation, differentiation, and production of a functional female gamete. The transzonal projections (TZP), which are specialized extensions from granulosa cells that terminate on the oolema after crossing the zona pellucida, are major structural components necessary for oocyte-GC interaction. Granulosa cell polarity seems to be a necessary requisite for appropriate function of TZP, and the role of FSH as modulator of a polarized phenotype on GC is discussed. This article also discusses oocyte polarity with special reference to the partial loss of polarity that occurs during in-vitro oocyte maturation and possible implications in the modulation of oocyte competencies. Cytoskeletal markers that may account for oocyte quality were defined and found to be distinct in in-vivo and in-vitro matured oocytes. Implications of partial loss of oocyte polarity during in-vitro maturation, reflected by distinct distribution of these markers, are further discussed. It is also proposed that expression of both somatic and germ cell polarity in the ovarian follicle will ultimately determine acquisition of meiotic, fertilization and developmental competences by the oocyte. PMID- 15901456 TI - Comparison of two sequential media for culturing cleavage-stage embryos and blastocysts: embryo characteristics and clinical outcome. AB - This study was undertaken to compare the outcome of day 3, day 5, and frozen thawed embryo transfer cycles where embryo culture was undertaken using the G1.2 G2.2 versus GIII series sequential media. A total of 400 day 3, 73 day 5 and 126 frozen-thawed embryo transfer cycles were analysed. Treatment cycles were quasi randomized on the day of oocyte retrieval to embryo culture in G1.2-G2.2 versus GIII series sequential media. Randomization was undertaken according to alternating weekdays. Significantly more embryos were of grade 1 or 2 quality on day 3 in the GIII group (P < 0.05). Likewise, more embryos in the GIII group had eight blastomeres on day 3 (P < 0.05) and were able to hatch spontaneously (P < 0.05). For day 3 embryo transfer cycles, implantation and clinical pregnancy rates were 14.4 and 25.7% versus 37.8 and 50.3% in G1.2-G2.2 and GIII groups respectively (P < 0.05). For day 5 embryo transfer cycles, implantation rates were significantly higher (29 versus 45%; P < 0.05) in the GIII group. There was a trend towards higher pregnancy rates; however, this did not reach statistical significance due to fewer cycles analysed in this group. In-vitro culture in GIII series sequential media yields better quality embryos that implant more efficiently, compared with culture in G1.2-G2.2 media. PMID- 15901457 TI - Is there still a place for co-cultures in the era of sequential media? AB - Co-cultures have been advocated in assisted reproduction owing to the inadequacy of simple media to support embryo development beyond the cleavage stage. Different human and non-human cells and cell lines have been used for co cultures. High rates of blastocyst formation have been reported with the use of co-cultures, and they have been proposed as a salvage treatment option in couples with repeated implantation failures. Since the advent of complex sequential media, which yield very high blastocyst formation and blastocyst implantation rates, the need for co-cultures has been questioned. Upon review of the literature, it is evident that well-designed randomized studies that compare co cultures with simple or sequential media do not exist. Progression to the blastocyst stage for cleavage stage embryos appears to be similar, if not better, for embryos that are cultured in modern sequential media, rendering the use of co cultures obsolete. Furthermore, there is no consensus regarding the necessity of sequential media, as similar results have been obtained with a single medium formulation that supports all stages of the preimplantation period. Whether co cultures are beneficial in patients with repeated implantation failures, however, should be investigated in randomized trials. Co-cultures still serve as powerful tools for understanding embryo metabolism. Furthermore, co-cultures may be instrumental in studying expression of implantation-related genes and embryo endometrium interaction. PMID- 15901458 TI - Differential staining combined with TUNEL labelling to detect apoptosis in preimplantation bovine embryos. AB - Development of accurate laboratory methods to assess embryo quality will improve the efficiency of embryo production from in-vitro culture systems. Currently, the techniques of TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end (TUNEL) labelling for the detection of apoptosis, and differential staining for determining the ratio of inner cell mass (ICM) to trophectoderm (TE) cells, are used separately to assess embryo quality in a range of different species. This paper reports a unique, simple and fast method for the assessment of embryo quality using differential staining of TE and ICM, but combined with TUNEL labelling (DST staining). This technique was used to investigate the effect of serum supplementation on total cell number, ICM:TE ratio and apoptosis index after in-vitro production of bovine embryos. Serum supplementation increased total cell number (P < 0.01), but reduced the ratio of ICM:TE cells. No differences were observed in the number of apoptotic nuclei between treatments, or in the localization of the apoptotic nuclei. However, more apoptotic nuclei were observed in ICM than TE cells in both culture groups. In conclusion, using DST, it has been possible to carry out both a qualitative and quantitative analysis of embryos produced using the two different methods. DST provides a means of assessing the effect of culture conditions on cell number of both embryo compartments (ICM and TE), as well as providing information on the localization of apoptotic nuclei within the blastocyst. PMID- 15901459 TI - Progressive elimination of microinjected trehalose during mouse embryonic development. AB - Recently, sugars such as trehalose have been introduced into mammalian cells by overcoming the permeability barrier of cell membranes, and have provided improved tolerance against stresses associated with freezing and drying. However, the fate of the intracellular sugars has remained an open question. To address this issue, mouse oocytes were microinjected with 0.1 mol/l trehalose, and intracellular trehalose and glucose concentrations were determined during embryonic development using a high performance liquid chromatography and pulsed amperometric detection protocol. Trehalose was not detected in non-injected controls at any stage of development. In the microinjection group, the amount of intracellular trehalose progressively decreased as embryos developed. There was a corresponding increase in intracellular glucose concentration at the two-cell stage, suggesting cleavage of trehalose to two glucose molecules. In summary, this study presents a simple, highly sensitive protocol to determine intracellular sugars. The data reveal rapid elimination of microinjected trehalose during embryonic development. These findings have implications for designing osmolarity-optimized culture media for sugar-injected oocytes. PMID- 15901460 TI - Granulocyte-macrophage colony-stimulating factor stimulates mouse blastocyst inner cell mass development only when media lack human serum albumin. AB - The aim of the current study was to examine the effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the development and differentiation of preimplantation mouse embryos from different strains and under different culture conditions. Embryos from F1 hybrid mice were cultured in a modified G1 medium lacking amino acids and EDTA (simple G1), human tubal fluid medium (HTF) or in G1/G2 sequential media, supplemented with GM-CSF (0, 2, 4, 8, and 16 ng/ml). Embryos from CF1 mice were subsequently cultured in G1/G2 with (5 mg/ml) or without HSA, in the absence or presence of GM-CSF (2 ng/ml). GM-CSF had no effect at any concentration on F1 embryo development and blastocyst cell numbers, irrespective of the culture media used. Similarly, GM-CSF had no effect on CF1 blastocyst development. However, a stimulatory effect of GM-CSF was evident on total blastocyst cell number and ICM development when CF1 embryos were cultured in the absence of HSA. When HSA was present in the media the beneficial effect of GM-CSF was negated. There was no difference in the number of apoptotic cells in CF1 blastocysts when G1/G2 were supplemented with GM-CSF with or without HSA. These data indicate that there is no beneficial effect of supplementing either simple (simple G1 or HTF) or more complete (G1/G2) media with GM-CSF when protein is present in the medium. However, when culture conditions are suboptimal and non physiological, i.e. the absence of protein, GM-CSF stimulates development of both total cell numbers and ICM development of CF1 blastocysts. PMID- 15901461 TI - Strategies for preimplantation genetic diagnosis of Angelman syndrome caused by mutations in the UBE3A gene. AB - Angelman syndrome (AS) is a neurodevelopmental disorder associated with the loss of maternal gene expression in chromosome region 15q11-q13. AS is caused by a wide variety of genetic mechanisms, including mutations in the UBE3A gene that have been identified in 10-15% of patients; when the mother is heterozygous for the causative mutation, the risk of recurrence in subsequent pregnancies is 50%. The present authors have developed a preimplantation genetic diagnosis (PGD) assay for a family displaying a 10 bp deletion in exon 9 of the UBE3A gene, which was shared by two affected children and their phenotypically normal mother. A duplex polymerase chain reaction protocol was established, allowing the efficient amplification of the mutation together with an informative microsatellite marker (D15S122) located in intron 1 of the UBE3A gene. As most of UBE3A mutations identified so far are unique to one family, the present authors have also developed an indirect single cell protocol based upon the co-amplification of two microsatellite markers located within (D15S122) and close to the UBE3A gene (D15S1506). This strategy may be applied to all informative families requesting PGD for Angelman syndrome associated with mutations in the UBE3A gene. PMID- 15901462 TI - Embryo-maternal interactive factors regulating the implantation process: implications in assisted reproductive. AB - The embryo-maternal dialogue that starts very early in the life of the embryo is crucial for its own implantation. A disturbance in this dialogue is the major reason for which 60% of all pregnancies are terminated at the end of the periimplantation period. Many studies have been performed to improve the understanding of the molecular mechanisms involved in this dialogue. Both partners, the mother and the embryo, are equally involved in this exchange of signals. Much progress has been done in understanding the role of (i) chorionic gonadotrophin, (ii) growth factors and cytokines, and (iii) steroid hormones and other mediators, produced either by the embryo, by the mother, or by both, during the peri-implantation period. Today it is clear that their production dictates changes in the endometrium, in the immunological system of the mother and in embryo metabolism, that enable the embryo to implant. Knowledge of the molecular mechanisms involved in the embryo-maternal interaction are reviewed in this article. PMID- 15901463 TI - Early detection of fetal structural abnormalities. AB - Most published data on the detection of fetal anomalies at 11-14 weeks are from specialized centres with considerable experience in fetal anomaly scanning. However, there is still limited information on the feasibility and limitations of the screening of these anomalies compared with the now classical mid-gestation screening. This review indicates that overall, the detection rate of fetal anomalies at 11-14 weeks is 44% compared with 74% by the mid-pregnancy scan. Major abnormalities of the fetal head, abdominal wall and urinary tract, and of the umbilical cord and placenta, can be reliably detected at 10-11 weeks of gestation. Detection of other anomalies such as spina bifida, diaphragmatic hernia or heart defects is limited before 13 weeks of gestation. So far it cannot be stated that routine first trimester screening can be used on a large scale to evaluate the fetal spine and heart in the general population. In particular, in screening for congenital heart defects, the ability to perform a full cardiac examination increases from 20% at 11 weeks to 92% at 13 weeks. The early prenatal diagnosis of these anomalies may be improved by screening at 13-14 weeks rather than during the first trimester. PMID- 15901470 TI - Allele frequencies and the r2 measure of linkage disequilibrium: impact on design and interpretation of association studies. AB - The design and interpretation of genetic association studies depends on the relationship between the genotyped variants and the underlying functional variant, often parameterized as the squared correlation or r(2) measure of linkage disequilibrium between two loci. While it has long been recognized that placing a constraint on ther(2) between two loci also places a constraint on the difference in frequencies between the coupled alleles, this constraint has not been quantified. Here, quantification of this severe constraint is presented. For example, for r(2) >/= .8, the maximum difference in allele frequency is +/- .06 which occurs when one locus has allele frequency .5. For r(2) >/= .8 and allele frequency at one locus of .1, the maximum difference in allele frequency at the second locus is only +/- .02. The impact on the design and interpretation of association studies is discussed. PMID- 15901471 TI - New concepts for distinguishing the hidden patterns of linkage disequilibrium which underlie association between genotypes and complex phenotypes. AB - A disappointing feature of conventional methods for detecting association between DNA variation and a phenotype of interest is that they tell us little about the hidden pattern of linkage disequilibrium (LD) with the functional variant that is actually responsible for the association. This limitation applies to case-control studies and also to the transmission/disequilibrium test (TDT) and other family based association methods. Here we present a fresh perspective on genetic association based on two novel concepts called 'LD squares' and 'equirisk alleles'. These describe and characterize the different patterns of gametic LD which underlie genetic association. These concepts lead to a general principle - the Equi-Risk Allele Segregation Principle - which captures the way in which underlying LD patterns affect the transmission patterns of genetic variants associated with a phenotype. This provides a basis for distinguishing the hidden LD patterns and might help to locate the functional variants responsible for the association. PMID- 15901472 TI - The value of four mental health self-report scales in predicting interview-based mood and anxiety disorder diagnoses in sibling pairs. AB - Questionnaire-based dimensional measures are often employed in epidemiological studies to predict the presence of psychiatric disorders. The present study sought to determine how accurately 4 dimensional mental health measures, the 12 item General Health Questionnaire (GHQ-12), Neuroticism (EPQ-N), the high positive affect and anxious arousal scales from the Mood and Anxiety Symptoms Questionnaire (MASQ-HPA and MASQ-AA) and a composite of all 4, predicted psychiatric caseness as diagnosed by the University of Michigan Composite International Diagnostic Interview (UM-CIDI). Community subjects were recruited through general practitioners; those who agreed to participate were sent a questionnaire containing the above measures. Subsequently, the UM-CIDI was administered by telephone to 469 subjects consisting of sibling pairs who scored most discordantly or concordantly on a composite index of the 4 measures. Logistic Regression and Receiver Operating Characteristic (ROC) curve analyses were carried out to assess the predictive accuracy of the dimensional measures on UM-CIDI diagnosis. A total of 179 subjects, 62 men and 117 women with an average age of 42 years, were diagnosed with at least one of the following psychiatric disorders: depression, dysthymia, generalized anxiety disorder (GAD), social phobia, agoraphobia and panic attack. The six disorders showed high comorbidity. EPQ-N and the Composite Index were found to be very strong and accurate predictors of psychiatric caseness; they were however unable to differentiate between specific disorders. The results from the present study therefore validated the four mental health measures as being predictive of psychiatric caseness. PMID- 15901473 TI - Where did I leave my keys? A twin study of self-reported memory ratings using the multifactorial memory questionnaire. AB - Behavior genetics has convincingly shown the importance of genetic factors in objective tests of memory function. However, self-report memory tests have received little attention. This study used items from the Multifactorial Memory Questionnaire (MMQ) to estimate the heritability of self-reported memory contentment and ability in 909 monozygotic (MZ) and 1034 dizygotic (DZ) twin pairs aged between 20 and 84 years from the St Thomas' Adult UK Twin Register. Heritability estimates ranged between 37% and 64% for contentment (e.g., reporting to worry about one's memory) and approximately 45% for ability (e.g., reporting a tendency to forget keys). Shared family environmental influences (between 32% and 33%) were found for some abilities (e.g., learning to use a new gadget). Given their clinical significance and ease of administration, these tests could prove to be useful in examining memory functioning in large-scale population studies. PMID- 15901474 TI - Comparative rating measures of health and environmental exposures: how well do twins agree? AB - Twins are sometimes used as proxy informants but little is known about reliability and validity of the information thus obtained. The present study asks: (1) to what extent do twin pairs agree with each other on comparative ratings of health, psychosocial traits, and environmental exposures?; and (2) how well do comparative ratings agree with usual self-reported information about the exposures? Using 55 monozygotic (MZ) and 71 dizygotic (DZ) same-sex pairs reared together, percentage agreement was calculated for 44 comparative ratings. Pairs agreed on average about half of the time. Agreement was higher for more discrete exposures, such as smoking, but lower for more subjective variables, such as the degree to which life is experienced as stressful. Signed rank tests were used to contrast comparative ratings to differences in self-reports. Differences between twin partners in their self-report indices, where available, were in the direction suggested by the comparative rating. Comparative ratings appear most accurate for smoking and alcohol use, and less consistent for mental health symptoms and self-rated health. PMID- 15901475 TI - Paternal alcoholism and offspring conduct disorder: evidence for the 'common genes' hypothesis. AB - Not only are alcoholism and externalizing disorders frequently comorbid, they often co-occur in families across generations; for example, paternal alcoholism predicts offspring conduct disorder just as it does offspring alcoholism. To clarify this relationship, the current study examined the 'common genes' hypothesis utilizing a children-of-twins research design. Participants were male monozygotic (MZ) and dizygotic (DZ) twins from the Vietnam Era Twin Registry who were concordant or discordant for alcohol dependence together with their offspring and the mothers of those offspring. All participants were conducted through a structured psychiatric interview. Offspring risk of conduct disorder was examined as a function of alcoholism genetic risk (due to paternal and co twin alcohol dependence) and alcoholism environmental risk (due to being reared by a father with an alcohol dependence diagnosis). After controlling for potentially confounding variables, the offspring of alcohol-dependent fathers were significantly more likely to exhibit conduct disorder diagnoses than were offspring of nonalcohol-dependent fathers, thus indicating diagnostic crossover in generational family transmission. Comparing offspring at high genetic and high environmental risk with offspring at high genetic and low environmental risk indicated that genetic factors were most likely responsible for the alcoholism conduct disorder association. The observed diagnostic crossover (from paternal alcoholism to offspring conduct disorder) across generations in the context of both high and low environmental risk (while genetic risk remained high) supported the common genes hypothesis. PMID- 15901476 TI - Human cytokine response to ex vivo amyloid-beta stimulation is mediated by genetic factors. AB - Through its ability to induce the enhanced release and production of cytokines, amyloid-beta is responsible for the chronic inflammatory response that contributes to Alzheimer's disease (AD). Determining whether the response of monocytes to amyloid-beta stimulation is under genetic control may help understand the basis of why some people are more prone to develop neuronal degeneration than others. In the current study we investigated the heritability of the cytokine (IL-10, IL-6, IL-1beta, IL-1ra, TNF-alpha) production capacity upon ex vivo stimulation with amyloid-beta in whole blood samples of 222 twins and 85 singleton siblings from 139 extended twin families. It was found that individual differences in amyloid-beta induced cytokine production capacity are to a large extent of genetic origin, with heritability estimates ranging from 55% (IL-1beta) to 68% (IL-6). We conclude that genes influencing amyloid-beta-induced cytokine response may provide clues to the progression of AD pathology. PMID- 15901477 TI - Genetic factors in seizures: a population-based study of 47,626 US, Norwegian and Danish twin pairs. AB - The purpose of the study was to describe a large sample of twins reporting a history of seizures, to characterize seizures in the three subpopulations, and to estimate the relative importance of genetic and environmental factors in seizure occurrence. Seizure history was determined by questionnaires completed by twins in population-based twin registries in the United States, Norway and Denmark. Concordance rates were calculated for all seizure categories within and across twin populations. Of 47,626 twin pairs evaluated, 6234 reported a history of seizures in one or both twins. Concordance rates were significantly higher for monozygotic (MZ) versus dizygotic (DZ)pairs for all seizure categories within and across populations. The results of this study involving the largest unselected, population-based sample of twins with seizures assembled to date confirm the importance of genetic factors in determining risk for epilepsy, febrile seizures, other seizures and staring spells. This sample is likely to provide an important resource for studying the genetics of epilepsy subtypes and febrile seizures. PMID- 15901478 TI - Birthweight and adult health in a population-based sample of Norwegian twins. AB - Population-based twin data were used to test (a) whether lower birthweight confers a greater risk of adult health disorders, and (b) whether within-pair birthweight differences in twins explain discordance for health outcomes. The sample consisted of 1201 monozygotic (MZ) male twins, 1048 dizygotic (DZ) male twins, 1679 MZ female twins, 1489 DZ female twins, and 2423 opposite-sex DZ twins, born in Norway between 1967 and 1979. The relationship between birthweight and self-reported health outcomes were studied using multivariable logistic regression. In the full sample (n = 7840), birthweight was negatively associated with risk for nearsightedness (odds ratio OR = 0.76, 95% CI: 0.65 - 0.92) and minimal brain disorder (OR = 0.27, 95% CI: 0.16-0.44) when adjusted for gestational age, sex, zygosity, age, education and body mass index after correction for intraclass correlations and multiple comparisons. Within-pair analysis of 159 MZ and 224 DZ pairs revealed that myopic twins were on average 2 g (p = .966) and 64 g (p = .040) lighter than nonmyopic twins in MZ and DZ pairs respectively, suggesting that genetic factors may play an important role in the associations between birthweight and nearsightedness. Within-pair analysis of twins discordant for a minimal brain disorder indicated that affected twins were 80 g (p = .655) and 85 g (p = .655) lighter than their healthy co-twins in MZ and DZ pairs respectively, although there were only 2 MZ and 2 DZ discordant pairs. PMID- 15901479 TI - The effects of twins, parity and age at first birth on cancer risk in Swedish women. AB - The effect of reproductive history on the risk of cervical, colorectal and thyroid cancers and melanoma has been explored but the results to date are inconsistent. We aimed to examine in a record-linkage cohort study the risk of developing these cancers, as well as breast, ovarian and endometrial cancers, among mothers who had given birth to twins compared with those who had only singleton pregnancies. Women who delivered a baby in Sweden between 1961 and 1996 and who were 15 years or younger in 1961 were selected from the Swedish civil birth register and linked with the Swedish cancer registry. We used Poisson regression to assess associations between reproductive factors and cancer. Twinning was associated with reduced risks of breast, colorectal, ovarian and uterine cancers, although no relative risks were statistically significant. The delivery of twins did not increase the risk of any cancers studied. Increasing numbers of maternities were associated with significantly reduced risks of all tumors except thyroid cancer. We found positive associations between a later age at first birth and breast cancer and melanoma, while there were inverse associations with cervix, ovarian, uterine and colorectal cancers. These findings lend weight to the hypothesis that hormonal factors influence the etiology of colorectal cancer in women, but argue against any strong effect of hormones on the development of melanoma or tumors of the thyroid. PMID- 15901480 TI - The convergence of the regional twinning rates in Sweden, 1751-1960. AB - Sweden has, as a whole nation, the oldest continuous population register of births, including twin and higher multiple maternities, starting in the 17th century. Until the 1920s, the rates of multiple maternities in Sweden were among the highest known among Europeans. Strong secular and regional fluctuations were noted. Some of the eastern counties showed especially marked decreasing trends. The twinning rate had no clear associations with the anthropometric and serological data. In this paper we study the temporal and regional variations of the twinning rate in the 25 counties of Sweden from 1751 to 1960. Different statistical methods were applied in order to test the hypothesis that, irrespective of the initial levels, the twinning rates for the counties converge towards a common low level. We present and interpret a geometrical model for the trends of the regional twinning rates. We also analyze the regional heterogeneity using the ranges and the coefficients of variation of the regional twinning rates. All the methods gave consistent results, supporting our convergence hypothesis that the regional differences in the twinning rate are gradually disappearing. In addition, this study supports our earlier findings that the regional heterogeneity cannot be explained by differences in the distribution of maternal age and parity. We suggest that the convergence may be caused by increased urbanization and industrialization and by the increased interregional migration of citizens as a consequence of better communications, which lead to the breaking up of isolates and decreased endogamy. PMID- 15901481 TI - Maternal serum steroid levels are unrelated to fetal sex: a study in twin pregnancies. AB - Increased prenatal exposure to testosterone (T) in females of an opposite-sex (OS) twin pair may have an effect on the development of sex-typical cognitive and behavioral patterns. The prenatal exposure to T due to hormone transfer in OS twin females may occur in two ways, one directly via the feto-fetal transfer route within the uterus, the other indirectly through maternal-fetal transfer and based in the maternal-fetal compartment. Although some studies in singletons indeed found that women pregnant with a male fetus have higher T levels during gestation than women pregnant with a female fetus, many other studies could not find any relation between the sex of the fetus and maternal serum steroid levels. Therefore at present it is unclear whether a pregnant woman bearing a male has higher levels of T than a woman bearing a female. Up to this point, no-one has investigated this issue in twin pregnancies. We examined the relationship between maternal serum steroid levels and sex of fetus in 17 female-female, 9 male-male and 29 OS twin pregnancies. No differences were observed between the maternal serum steroid levels of women expecting single-sex and mixed-sex offspring. It is concluded that the source of prenatal T exposure in females probably comes from the fetal unit, which is the direct route of fetal hormone transfer. PMID- 15901482 TI - Chimeric dizygotic twins: implications for development and disease. PMID- 15901483 TI - Altered distribution of striatal activity-dependent synaptic plasticity in the 3 nitropropionic acid model of Huntington's disease. AB - Huntington's disease (HD) is a neurodegenerative disorder characterized by involuntary choreiform movements, neuropsychiatric disturbances and cognitive decline. The hyperkinetic phenomenology has commonly been attributed to a disturbance of the basal ganglia function, mainly the neostriatum, but its pathophysiology mechanisms remain unclear. Activity-dependent long-term changes in synaptic efficacy, such as long-term potentiation (LTP) and long-term depression (LTD), are widely considered to be the cellular models for acquisition and storage of information in neuronal networks. Both LTP and LTD have been described at the corticostriatal pathway and they might be probably involved not only in physiological motor behavior processing but also in disease states affecting that pathway. Systemic injection of 3-nitropropionic acid (3-NP) induces excitotoxic striatal lesions and abnormal movements in rodents, resembling those seen in HD. We examined synaptic plasticity in dorsolateral striatum slices prepared from both control and 3-NP-treated rats by recording extracellular field potentials. Our results reinforce the idea that both forms of activity-dependent synaptic plasticity can be recorded at the dorsolateral region of striatum by the same stimulating protocol in control rats and suggest that 3 NP-induced striatal lesions may be associated with suppression of LTD expression in the sensorimotor striatum. PMID- 15901484 TI - Tetranectin in cerebrospinal fluid: biochemical characterisation and evidence of intrathecal synthesis or selective uptake into CSF. AB - BACKGROUND: Tetranectin (TN) is a 67 kDa glycoprotein thought to play a prominent role in the regulation of proteolytic processes via its binding to plasminogen and indirect activation of plasminogen. The TN concentration in serum is approximately 10 mg/l and is reduced in patients with several cancers. The TN concentration in the normal CSF has not been examined. METHODS: The TN concentration in the serum and CSF of 47 normal subjects without neurological disorders was established using a polyclonal sandwich ELISA. RESULTS: The median TN concentration (quartile range) was 10.8 mg/l (9.0-12.1) in serum and 0.43 mg/l (0.3-0.53) in CSF. The TN index median (quartile range), defined as (TN CSF concentration/TN serum concentration)/(Albumin CSF concentration/Albumin serum concentration), was found to be 5.5 (4.7-7.6), suggesting intrathecal synthesis or selective uptake of TN in CNS. Immunohistochemistry showed TN immunoreactivity in neurons and dendrites, but no staining in glial cells in the cerebrum and cerebellum. In plexus choroideus, the ependymal cells exhibited strong immunoreactivity. TN in serum and CSF were immunochemically identical and of similar size. CONCLUSION: TN is present in normal brain and CSF, and the TN index is very high, but further studies are necessary to decide whether TN is synthesised in the CNS or selectively transported over the blood-brain barrier. PMID- 15901485 TI - Sca-1 negatively regulates proliferation and differentiation of muscle cells. AB - Satellite cells are tissue-specific stem cells critical for skeletal muscle growth and regeneration. Upon exposure to appropriate stimuli, satellite cells produce progeny myoblasts. Heterogeneity within a population of myoblasts ensures that a subset of myoblasts readily differentiate to form myotubes, whereas other myoblasts remain undifferentiated and thus available for future muscle growth. The mechanisms that contribute to this heterogeneity in myoblasts are largely unknown. We show that satellite cells are Sca-1(neg) but give rise to myoblasts that are heterogeneous for sca-1 expression. The majority of myoblasts are sca 1(neg), rapidly divide, and are capable of undergoing myogenic differentiation to form myotubes. In contrast, a minority population is sca-1(pos), divides slower, and does not readily form myotubes. Sca-1 expression is not static but rather dynamically modulated by the microenvironment. Gain-of-function and loss-of function experiments demonstrate that sca-1 has a functional role in regulating proliferation and differentiation of myoblasts. Myofiber size of sca-1 null muscles is altered in an age-dependent manner, with increased size observed in younger mice and decreased size in older mice. These studies reveal a novel system that reversibly modulates the myogenic behavior of myoblasts. These studies provide evidence that, rather than being a fixed property, myoblast heterogeneity can be modulated by the microenvironment. PMID- 15901486 TI - 4-Hydroxyestradiol induces oxidative stress and apoptosis in human mammary epithelial cells: possible protection by NF-kappaB and ERK/MAPK. AB - Catechol estrogens, the hydroxylated metabolites of 17beta-estradiol (E2), have been considered to be implicated in estrogen-induced carcinogenesis. 4 Hydroxyestradiol (4-OHE2), an oxidized metabolite of E2 formed preferentially by cytochrome P450 1B1, reacts with DNA to form depurinating adducts thereby exerting genotoxicity and carcinogenicity. 4-OHE2 undergoes 2-electron oxidation to quinone via semiquinone, and during this process, reactive oxygen species (ROS) can be generated to cause DNA damage and cell death. In the present study, 4-OHE2 was found to elicit cytotoxicity in cultured human mammary epithelial (MCF 10A) cells, which was blocked by the antioxidant trolox. MCF-10A cells treated with 4-OHE2 exhibited increased intracellular ROS accumulation and 8-oxo-7,8 dihydroxy-2'-deoxyguanosine formation, and underwent apoptosis as determined by poly(ADP-ribose)polymerase cleavage and disruption of mitochondrial transmembrane potential. The redox-sensitive transcription factor nuclear factor kappaB (NF kappaB) was transiently activated by 4-OHE2 treatment. Cotreatment of MCF-10A cells with the NF-kappaB inhibitor, L-1-tosylamido-2-phenylethyl chloromethyl ketone, exacerbated 4-OHE2-induced cell death. 4-OHE2 also caused transient activation of extracellular signal-regulated protein kinases (ERK) involved in transmitting cell survival or death signals. A pharmacological inhibitor of ERK aggravated the 4-OHE2-induced cytotoxicity, supporting the pivotal role of ERK in protecting against catechol estrogen-induced oxidative cell death. PMID- 15901487 TI - Isolation and characterization of a new Vesivirus from rabbits. AB - This report describes the isolation, cDNA cloning, complete genome nucleotide sequence, and partial characterization of a new cultivable calicivirus isolated from juvenile feeder European rabbits (Oryctolagus cuniculus) showing symptoms of diarrhea. Absence of neutralization by type-specific neutralizing antibodies for 40 caliciviruses and phylogenetic sequence comparisons of the open reading frame 1-encoded polyprotein with those of other caliciviruses demonstrate that this new calicivirus is a putative novel member of the Vesivirus genus which is closely related to the marine calicivirus subgroup. According to its putative classification, this new virus has been named rabbit vesivirus. PMID- 15901488 TI - Seasonal variations and mass closure analysis of particulate matter in Hong Kong. AB - The chemical characteristics of ambient particulate matters in urban and rural areas of Hong Kong were determined in this study. A monitoring program starting from November 2000 to February 2001 (winter) and June 2001 to August 2001 (summer) for PM10 and PM2.5 was performed at three monitoring stations in Hong Kong. Twenty-four-hour PM10 and PM2.5 samples were collected once every 6 days at two urban sites, PolyU and KT, and every 12 days at a background site, HT, with Hi-Vol samplers. High concentrations of OC, EC (except in PolyU), water-soluble ions and elements were observed in winter among the three sampling sites for PM10 and PM2.5 fractions. Seasonal variations were significant in background HT. Dilution effect due to the increase in mixing depth and precipitation in summer reduced the concentrations of particulate matters. Long-range transport could contribute to the higher concentrations of particulate matter in the winter. Chemical mass closure calculations were performed for PM10 and PM2.5 observed. Mass closure improved when separate factors (1.4 and 1.9 respectively) were used to convert water-soluble organic carbon (WSOC) and water-insoluble organic carbon (WINSOC) into corresponding organic masses. The urban sites showed high percentages of water-soluble ions in winter and high percentages of carbonaceous species in summer. Better results were obtained for the chemical mass closure analysis in winter than in summer. High temperature and solar radiation in summer increased the rate of the complex photochemical reaction in the atmosphere. Therefore the chemical mass closure analysis would underestimate the volatized species and secondary aerosols during summer. PMID- 15901489 TI - Human netrin-G1 isoforms show evidence of differential expression. AB - The recently identified netrins-G1 and -G2 form a distinct subgroup within the UNC-6/netrin gene family of axon guidance molecules. In this study, we determined the size and structure of the exon/intron layout of the human netrin-G1 (NTNG1) and -G2 (NTNG2) genes. Northern analysis of both genes showed limited nonneuronal but wide brain expression, particularly for NTNG2. Reverse transcriptase PCR detected nine alternatively spliced isoforms including four novel variants of NTNG1 from adult brain. A semiquantitative assay established that major expression was restricted to isoforms G1c, G1d, G1a, and G1e in the brain and to G1c in the kidney. There is also evidence of developmental regulation of these isoforms between fetal and adult brain. In conclusion, NTNG1 may use alternative splicing to diversify its function in a developmentally and tissue-specific manner. PMID- 15901490 TI - Imaging phonology without print: assessing the neural correlates of phonemic awareness using fMRI. AB - Acquisition of phonological processing skills, such as the ability to segment words into corresponding speech sounds, is critical to the development of efficient reading. Prior neuroimaging studies of phonological processing have often relied on auditory stimuli or print-mediated tasks that may be problematic for various theoretical and empirical reasons. For the current study, we developed a task to evaluate phonological processing that used visual stimuli but did not require interpretation of orthographic forms. This task requires the subject to retrieve the names of objects and to compare their first sounds; then, the subject must indicate if the initial sounds of the names of the pictures are the same. The phonological analysis task was compared to both a baseline matching task and a more complex control condition in which the participants evaluated two different pictures and indicated whether they represented the same object. The complex picture-matching condition controls for the visual complexity of the stimuli but does not require phonological analysis of the names of the objects. While both frontal and ventral posterior areas were activated in response to phonological analysis of the names of pictures, only inferior and superior frontal gyrus exhibited differential sensitivity to the phonological comparison task as compared to the complex picture-matching control task. These findings suggest that phonological processing that is not mediated by print relies primarily on frontal language processing areas among skilled readers. PMID- 15901492 TI - Cotranscriptional mRNP assembly: from the DNA to the nuclear pore. AB - Transcription is coupled with the concomitant assembly of RNA-binding proteins to the nascent mRNA to generate a stable and export-competent mRNP particle. RNA binding factors recruited at active transcription sites specify the processing, nuclear export, subcellular localization, translation and stability of the mRNA. The assembly of the mRNP particle starts with the association of the cap-binding protein complex followed by the splicing-dependent assembly of the exon-junction complex in intron-containing genes and by the binding of RNA-export adaptor proteins. New findings suggest that mRNP assembly is a genetically controlled process that plays a key role in gene expression and other cellular processes, including the maintenance of genome integrity. PMID- 15901493 TI - Rules of engagement: co-transcriptional recruitment of pre-mRNA processing factors. AB - The universal pre-mRNA processing events of 5' end capping, splicing, and 3' end formation by cleavage/polyadenylation occur co-transcriptionally. As a result, the substrate for mRNA processing factors is a nascent RNA chain that is being extruded from the RNA polymerase II exit channel at 10-30 bases per second. How do processing factors find their substrate RNAs and complete most mRNA maturation before transcription is finished? Recent studies suggest that this task is facilitated by a combination of protein-RNA and protein-protein interactions within a 'mRNA factory' that comprises the elongating RNA polymerase and associated processing factors. This 'factory' undergoes dynamic changes in composition as it traverses a gene and provides the setting for regulatory interactions that couple processing to transcriptional elongation and termination. PMID- 15901494 TI - Connections between mRNA 3' end processing and transcription termination. AB - Discoveries within the last few years have revealed that the multiple steps in gene expression are remarkably integrated. There have recently been several advances in deciphering how mRNA 3' end processing is linked with transcription elongation and termination. It has been known for quite a long time that transcription termination is somehow intertwined with polyadenylation, but it is still unclear exactly how these two processes influence each other. Some recent reports are shedding light on these connections. PMID- 15901495 TI - Promoter usage and alternative splicing. AB - Recent findings justify a renewed interest in alternative splicing (AS): the process is more a rule than an exception as it affects the expression of 60% of human genes; it explains how a vast mammalian proteomic complexity is achieved with a limited number of genes; and mutations in AS regulatory sequences are a widespread source of human disease. AS regulation not only depends on the interaction of splicing factors with their target sequences in the pre-mRNA but is coupled to transcription. A clearer picture is emerging of the mechanisms by which transcription affects AS through promoter identity and occupation. These mechanisms involve the recruitment of factors with dual functions in transcription and splicing (i.e. that contain both functional domains and hence link the two processes) and the control of RNA polymerase II elongation. PMID- 15901496 TI - TREX, SR proteins and export of mRNA. AB - The machineries involved in gene expression are highly conserved from yeast to metazoans. However, a fundamental difference between these organisms is that most yeast genes lack introns whereas the converse is true in higher organisms. Recent studies of the TREX complex, which functions in mRNA export, unexpectedly revealed that this complex is recruited by the transcription machinery in yeast whereas the TREX complex appears to be recruited by the splicing machinery in mammals. Studies during the past year also revealed a possible conserved role for SR protein dephosphorylation in regulating the interaction between SR proteins and the mRNA export receptor TAP (Mex67 in yeast). There is also an interesting possibility that an SR protein-TREX complex interaction is a conserved part of the mRNA export machinery. PMID- 15901497 TI - Metazoan replication-dependent histone mRNAs: a distinct set of RNA polymerase II transcripts. AB - Metazoan replication-dependent histone mRNAs are the only eukaryotic mRNAs that lack polyA tails. The genes for the five histone proteins have remained physically linked during evolution. Expression of histone mRNAs and histone proteins requires a unique set of factors, and may be coordinated by association of the histone genes with Cajal bodies. Recently several novel factors, including components of the U7 snRNP, as well as proteins involved in regulation of histone gene expression, have been described. PMID- 15901498 TI - Crosstalk in gene expression: coupling and co-regulation of rDNA transcription, pre-ribosome assembly and pre-rRNA processing. AB - Ribosomes, the large RNPs that translate mRNA into protein in the cytoplasm of eukaryotic cells, are synthesized in a subcompartment of the nucleus, the nucleolus. There, transcription by Pol I yields a pre-rRNA which is modified, cleaved and assembled with ribosomal proteins to make functional ribosomes. Previously, rRNA transcription and pre-rRNA cleavage in eukaryotes were considered to be separable steps in gene expression. However, recent findings suggest that these two steps in gene expression can be concurrent and are co regulated. Unexpectedly, optimal rDNA transcription requires the presence of a defined subset of components of the pre-rRNA processing machinery. PMID- 15901499 TI - Formation of export-competent mRNP: escaping nuclear destruction. AB - In eukaryotic cells, primary transcripts are processed and bound by proteins before export to the cytoplasm. Nuclear production of export-competent messenger ribonucleoprotein particles (mRNPs) is a complicated process, and mRNP biogenic events that function sub-optimally are rapidly attacked by surveillance leading to degradation of the mRNA. Export of nuclear mRNAs is therefore constantly challenged by the opposing force of mRNA retention and decay. This balance ensures that only 'perfect' transcripts persist, and that non-functional and potentially deleterious transcripts are removed early in their biogenesis. Thus, eukaryotic systems of mRNP quality control can be viewed as simple Darwinian principles operating at the molecular level. PMID- 15901500 TI - Quality control of messenger ribonucleoprotein particles in the nucleus and at the pore. AB - The spatial separation of nuclear transcription and cytoplasmic translation in eukaryotic cells implies that mRNAs have to travel. On their journey, proteins involved in the various steps of transcript formation, processing and transport dynamically interact with mRNAs to form diverse messenger ribonucleoprotein complexes (mRNPs). Increasing evidence indicates that the protein complexes involved in distinct phases of manufacturing a bona fide mRNA in the nucleus are tightly coupled. Moreover, the recent demonstration that active genes migrate into preassembled, shared nuclear sub-compartments suggests that mRNAs are churned out in large 'transcription factories' with distinct but interconnected divisions. Nuclear factors have now been identified that specifically control the quality of mRNAs without affecting mRNP biogenesis or export. PMID- 15901501 TI - Retention and repression: fates of hyperedited RNAs in the nucleus. AB - Double-stranded RNA (dsRNA) is often formed in the nuclei of mammalian cells, but in this compartment it does not induce the effects characteristic of cytoplasmic dsRNA. Rather, recent work has suggested that nuclear dsRNA is a target for the ADAR class of enzymes, which deaminate adenosines to inosines. Further, there are a number of distinct fates of such edited RNA, including nuclear retention and perhaps also gene silencing. PMID- 15901502 TI - Mechanistic links between nonsense-mediated mRNA decay and pre-mRNA splicing in mammalian cells. AB - Nonsense-mediated mRNA decay (NMD) generally involves nonsense codon recognition by translating ribosomes at a position approximately 25 nts upstream of a splicing-generated exon junction complex of proteins. As such, NMD provides a means to degrade abnormal mRNAs that encode potentially deleterious truncated proteins. Additionally, an estimated one-third of naturally occurring, alternatively spliced mRNAs is also targeted for NMD. Given the extraordinary frequency of alternative splicing together with data indicating that naturally occurring transcripts other than alternatively spliced mRNAs are likewise targeted for NMD, it is believed that mammalian cells routinely utilize NMD to achieve proper levels of gene expression. PMID- 15901503 TI - Nonsense-mediated mRNA decay: molecular insights and mechanistic variations across species. AB - Nonsense-mediated mRNA decay (NMD) is an mRNA surveillance pathway that ensures the rapid degradation of mRNAs containing premature translation termination codons (PTCs), thereby preventing the synthesis of truncated and potentially harmful proteins. In addition, this pathway regulates the expression of approximately 10% of the transcriptome and is essential in mice. Although NMD is conserved in eukaryotes, recent studies in several organisms have revealed that different mechanisms have evolved to discriminate natural from premature stop codons and to degrade the targeted mRNAs. With the elucidation of the first crystal structures of components of the NMD machinery, the way is paved towards a molecular understanding of the protein interaction network underlying this process. PMID- 15901504 TI - RNA decapping inside and outside of processing bodies. AB - Decapping is a central step in eukaryotic mRNA turnover. Recent studies have identified several factors involved in catalysis and regulation of decapping. These include the following: an mRNA decapping complex containing the proteins Dcp1 and Dcp2; a nucleolar decapping enzyme, X29, involved in the degradation of U8 snoRNA and perhaps of other capped nuclear RNAs; and a decapping 'scavenger' enzyme, DcpS, that hydrolyzes the cap structure resulting from complete 3'-to-5' degradation of mRNAs by the exosome. Several proteins that stimulate mRNA decapping by the Dcp1:Dcp2 complex co-localize with Dcp1 and Dcp2, together with Xrn1, a 5'-to-3' exonuclease, to structures in the cytoplasm called processing bodies. Recent evidence suggests that the processing bodies may constitute specialized cellular compartments of mRNA turnover, which suggests that mRNA and protein localization may be integral to mRNA decay. PMID- 15901507 TI - Driving in America: a convergence of public policy and science. PMID- 15901505 TI - Dynamics of transcription and mRNA export. AB - Understanding the different molecular mechanisms responsible for gene expression has been a central interest of molecular biologists for several decades. Transcription, the initial step of gene expression, consists of converting the genetic code into a dynamic messenger RNA that will specify a required cellular function following translocation to the cytoplasm and translation. We now possess an in-depth understanding of the mechanism and regulations of transcription. By contrast, an understanding of the dynamics of an individual gene's expression in real time is just beginning to emerge following recent technological developments. PMID- 15901509 TI - The year 2004: a depressing year for the treatment of depression in adolescence. PMID- 15901510 TI - Protection against antisocial behavior in children exposed to physically abusive discipline. AB - PURPOSE: The study investigated protective factors (school commitment/importance, parent/peer disapproval of antisocial behavior, positive future orientation, and religion) hypothesized to lower risk for antisocial behavior among adolescents who, as children, had been physically abused. Protective factors also were investigated for comparison, nonabused children, and for children at risk on abuse and other factors: low socioeconomic status and early antisocial behavior. METHODS: Analyses used a two-step hierarchical regression approach. In step 1, age, gender, and early antisocial behavior were entered as controls. In step 2, each protective factor was entered separately as a predictor. A final regression model in each case examined the additive (combined) effect of all protective factors on a given outcome. Tests of predictor-by-group interactions were used to examine group differences. RESULTS: Among abused and nonabused children, having a strong commitment to school, having parents and peers who disapprove of antisocial behavior, and being involved in a religious community lowered rates of lifetime violence, delinquency, and status offenses. Having a positive future orientation appeared less powerful as a protective influence. Exposure to an increasing number of protective factors was for each outcome associated with a diminution in risk for antisocial behavior. CONCLUSIONS: Protective factors represent targets for preventive intervention that are viable for children as they enter adolescence. The fact that protective factors were predictive of lower antisocial behavior in both the abuse and comparison groups suggests that protective effects are more universal than they are unique to a given group of children. PMID- 15901511 TI - Does perceived parental rejection make adolescents sad and mad? The association of perceived parental rejection with adolescent depression and aggression. AB - PURPOSE: To research the association of perceived parental rejection to adolescent depression and aggression. METHODS: This study focused on 1329 Dutch junior high and high school students (47.9% males and 52.1% females; age range 10 19 years) that completed depression, aggression and perceived parental rejection questionnaires. The data were analyzed by structural equation modeling that assumed a relationship between perceived parental rejection and adolescent aggression, as mediated by adolescent depression. RESULTS: Perceived parental rejection, mediated through adolescent depression, explains aggressive behaviors of adolescents, as tested by a mediation model. Additionally, the fit of this mediation model is somewhat enhanced when direct paths from perceived parental rejection to aggression are included. Further analysis demonstrates that these effects are also somewhat dependent on the gender and the age of the adolescents, as would be expected in light of previous studies of these cohorts. CONCLUSIONS: The study of perceived parental rejection should receive the same attention in the research of the development of both adolescent depression and aggression, as has been the case for adolescent peer rejection. PMID- 15901512 TI - The influence of smoking imagery on the smoking intentions of young people: testing a media interpretation model. AB - PURPOSE: To assess a theoretical model of adolescents' exposure to films, perceptions of smoking imagery in film, and smoking intentions. METHODS: A structured questionnaire was completed by 3041 Year 8 (aged 12 years) and Year 12 (aged 16 years) students from 25 schools in Auckland, New Zealand. The survey assessed the relationships among exposure to films, attitudes about smoking imagery, perceptions of smoking prevalence and its acceptability, and expectations of smoking in the future. Measures included exposure to films, perceived pervasiveness of, and nonchalant attitudes about smoking imagery, identification of positive smoker stereotypes in films, perceived smoking prevalence, judgment of smoking acceptability, and smoking expectations. Path analytic techniques, using multiple regression analyses, were used to test the pattern of associations identified by the media interpretation model. RESULTS: Hierarchical regression analyses revealed that film exposure predicted higher levels of perceived smoking prevalence, perceived imagery pervasiveness, and nonchalant attitudes about smoking imagery. Nonchalant attitudes, identification of positive smoker stereotypes, and perceived smoking prevalence predicted judgments of smoking acceptability. Acceptability judgments, identification of positive stereotypes, and perceived smoking prevalence were all positively associated with smoking expectations. The media interpretation model accounted for 24% of the variance in smoking expectations within the total sample. CONCLUSIONS: Smoking imagery in film may play a role in the development of smoking intentions through inflating the perception of smoking prevalence and presenting socially attractive images. PMID- 15901513 TI - Sport and physical activity participation and substance use among adolescents. AB - PURPOSE: To examine the association between participation in specific school sponsored sports and out-of-school sports/physical activities and substance use. METHODS: Subjects consisted of 891 8th grade youth from three schools. Baseline data were collected using the Youth Alcohol & Drug Survey (2000) and following standardized protocols. Logistic regressions were conducted to identify associations between the independent variables of school-sponsored sports, and out-of-school sports/physical activities, and each of the four substance use dependent variables, while controlling for race. Additionally, logistic regressions were run separately for males and females to examine gender differences. RESULTS: Participation in any one of seven specific sports/physical activities was associated with increased substance use for one or both genders, whereas participation in any one of four other specific sports/physical activities was associated with decreased use for one or both genders. Those sports associated with increased use differed for males and females, as did those associated with decreased use. Females in school-sponsored dance/cheerleading/gymnastics were at decreased risk of alcohol use, whereas those in out-of-school dance/cheerleading/gymnastics, skateboarding or surfing were at increased risk for using at least one substance. Males in out-of-school swimming were at decreased risk of heavy alcohol use, whereas those in school sponsored football, swimming, wrestling or out-of-school tennis were at increased risk for using at least one substance. CONCLUSIONS: Educators cannot assume all sports/physical activities have a positive relationship with youth substance use. School-sponsored, male-dominated sports appeared to be associated with an increased substance use risk for males, whereas out-of-school, mixed-gender sports appeared to be for females. PMID- 15901514 TI - Fruits, vegetables, and football: findings from focus groups with alternative high school students regarding eating and physical activity. AB - PURPOSE: To increase our understanding of factors that may influence the dietary and physical activity practices of adolescents attending an alternative high school (AHS). METHODS: Seventy students (36 girls, 34 boys) from urban and suburban AHSs in the Minneapolis-St. Paul metropolitan area participated in 7 focus groups to discuss their perceptions and opinions about factors that influence their eating and physical activity behaviors and to offer suggestions regarding school-based strategies to support and to promote healthy physical activity and eating practices among students. Mixed-gender groups were facilitated by a trained moderator by using a set of standardized questions to guide the 45- to 60-minute discussions. Focus groups were audiotaped, transcribed, and analyzed using a 3-step process for qualitative analysis. RESULTS: Time, cost, availability, and convenience were identified as key factors that influenced students' food choices and the choice to be active physically. Access to healthy foods and physical activity was problematic, especially at school. Students also reported that social support from their friends, family, and teachers, and role-modeling behaviors of adults enhanced their likelihood of eating healthy foods and being active. CONCLUSIONS: Study findings suggest that programs that target social-environmental factors that include norms, role models, social support, and opportunities to practice a health behavior have the potential to affect positively the dietary and physical activity practices of teenagers attending an AHS. Interventions that aim to increase opportunities at school to practice healthy eating and physical activity may be effective, especially in promoting and supporting healthy behavior change among students. PMID- 15901515 TI - Comparison of serum lipid levels among Korean, Korean-Chinese, and Han-Chinese adolescents. AB - PURPOSE: To follow offspring of emigrants from Korea to China to assess the effects of obesity and urbanization/westernization on atherogenic risk profiles. Obesity and serum lipid levels during adolescence are associated with risk for atherosclerotic diseases during adulthood, but the effect of obesity on serum lipid levels in relatively lean populations is unclear, particularly among adolescents. METHODS: The correlation of anthropometric measures with serum lipids was assessed in 2345 adolescents aged 16 to 18 years (four study groups: Korean-Chinese, n = 701; Korean-Rural, n = 671; Korean-Urban, n = 523; and Han Chinese, n = 450). RESULTS: Korean adolescents had higher average total cholesterol (TC) levels (146 mg/dl and 156 mg/dl for boys and girls, respectively) compared with Chinese adolescents (127 mg/dl and 143 mg/dl in China, respectively). The racial difference in atherogenic lipids (TC, low density lipoprotein cholesterol [LDL-C], or their ratio to high-density lipoprotein cholesterol [HDL-C]) between Korean-Chinese and Han-Chinese was not found, but the urbanization difference among Korean adolescents (Korean-Chinese, Korean-Rural, and Korean-Urban) was obvious. The correlations of serum lipid levels with obesity indices were significant in TC, LDL-C, TC/HDL-C, LDL-C/HDL-C, and triglyceride (TG) among boys (p < .001) and in HDL-C, TC/HDL-C, LDL-C/HDL-C, and TG among girls (p < .05), but not strongly correlated (Pearson r < .2). CONCLUSIONS: Significantly higher levels of TC and LDL-C (approximately 20 mg/dl) in Korean adolescents compared with Chinese adolescents were not owing to obesity or any racial difference. These findings underscore the importance of health promotion strategies including changes of lifestyle during childhood and adolescence. PMID- 15901516 TI - Is peer education the best approach for HIV prevention in schools? Findings from a randomized controlled trial. AB - PURPOSE: To evaluate the effectiveness of peer education when compared to teacher led curricula in AIDS prevention programs conducted in schools in Rome, Italy. METHODS: Eighteen high schools were randomly assigned to one of two prevention programs: one led by teachers and the other by peer leaders. A sample of students attending the last 2 years (n = 1295) completed pre and post-intervention questionnaires. Changes in sexual behaviors, knowledge, prevention skills, risk perception and attitudes were first evaluated within each intervention group. Afterwards, changes in knowledge, prevention skills, risk perception, and attitudes total scale scores (post-test scores minus pre-test scores) were compared between the two groups, through linear regression models, in order to control for confounders, taking into account the within-school correlation in score changes. As for sexual behaviors, number of sexual partners and frequency of condom use in the 3 months before post-test were compared by intervention group through ordinal regression models, also taking into account behaviors reported before pre-test. RESULTS: For both groups, we observed significant improvements in skills, knowledge, attitudes and risk perception. The peer-led group showed a 6.7% (95% C.I. 1.9-11.5) scores greater improvement in knowledge, compared to the teacher-led group. In neither group were improvements observed in condom use or number of sexual partners. CONCLUSIONS: The only apparent benefit of the peer-led intervention, compared to that led by teachers, was a greater improvement in knowledge of HIV. Neither of the interventions induced changes in sexual behavior. However, the role of possible biases and methodological problems must be considered when interpreting these results. PMID- 15901517 TI - Adolescent cigarette smoking and mental health symptoms. AB - PURPOSE: Adolescent cigarette smoking has been reported to be associated with substance use and depression. Less is known about the association between smoking and other mental health symptoms, or the impact of mental health treatment on cigarette smoking and psychiatric morbidity including the substance use disorders. The purpose of this study is to examine the association among adolescent cigarette smoking, mental health symptoms including substance use disorders, and mental health treatment among adolescents treated at a large, hospital-based young adult clinic. METHODS: Observational study of 486 adolescent patients presenting for routine medical care. RESULTS: Participants were two thirds female with an average age of 16.6 years and of diverse ethnic background; 129 (28.6%) reported ever-using tobacco products, and 68 (14%) were current smokers who had an average of 2 cigarettes per day. Current smoking was associated with significantly increased odds of having mental health symptoms and substance use disorders, even after controlling for age and previous mental health treatment, and in both boys and girls. CONCLUSIONS: This study supports the increasing appreciation that adolescent tobacco use is clinically important, even at apparently low levels. Adolescent cigarette smoking is generally more common than alcohol and illicit drug use, and these results highlight its multifarious linkages with use of other substances and mental health symptoms. Thus, early identification and intervention of adolescent cigarette smoking may help not only to avert later daily smoking, but also may assist in identifying patients at increased risk for substance use and psychiatric problems. PMID- 15901518 TI - Does office-based counseling of adolescents and young adults improve self reported safety habits? A randomized controlled effectiveness trial. AB - PURPOSE: The purpose of this study was to evaluate brief physician advice regarding seatbelt and bicycle helmet use in adolescents and young adults. METHODS: We recruited 200 patients ages 11-24 years presenting for all visits to a primary care clinic in the Southwestern United States from January 2000 to March 2001. Patients were randomized to control or a single 2- to 3-minute scripted motivational counseling intervention delivered by physicians with an educational brochure and discount helmet coupon. We conducted telephone follow-up evaluation at 3 months. Main outcome measures were self-reported seatbelt and bicycle helmet use assessed on a 4-point Likert scale and attitudes toward these behaviors assessed on a 5-point Likert scale and analyzed by the Wilcoxon signed rank test. RESULTS: For the intervention group, mean Likert scores precounseling and postcounseling for self-reported seatbelt use were 1.3 and 1.4, respectively, with a mean difference of .04 (95% confidence interval [CI], -.1 to .2). For self reported bicycle helmet use, scores were 3.1 and 3.0, respectively, with a mean difference of .1 (95% CI, -.3 to .2). Combined mean Likert scores measuring subject's attitudes about seat belt use were 13.8 and 14.0, respectively, with a mean difference of .2 (95% CI, -.3 to .6). For bicycle helmet use, scores were 20.0 and 20.9, respectively, with a mean difference of .9 (95% CI, -1.4 to 3.3). CONCLUSION: We were unable to detect a significant improvement in safety behaviors or attitudes by adolescents and young adults after a brief intervention by physicians during routine office visits. PMID- 15901519 TI - Which pediatric residents assist and arrange follow-up for patients and parents who use tobacco? AB - The current study seeks to identify predictors of pediatric resident tobacco intervention behaviors. Training in the residency program, beliefs about the appropriateness of pediatric interventions, and beliefs about the availability of effective interventions were most likely to predict intervention with assisting and arranging strategies. PMID- 15901520 TI - Condom failure among adolescents: implications for STD prevention. AB - This study of 921 adolescents found condom failure (past 90 days) was experienced by at least one-third of the sample, regardless of gender. Frequency of condom failure was positively associated with STD diagnosis (AOR = 1.22, 95% CI = 1.01 1.48), with the odds of testing positive increasing 22% for each added event of failure. PMID- 15901521 TI - Classifying asthma severity: prospective symptom diary or retrospective symptom recall? AB - We used an observational within-subject control study at an inner-city school to compare asthma severity classification obtained by retrospective symptom recall with that obtained by a prospective symptom diary. The prospective symptom diary identified a higher proportion of persistent asthmatics than the retrospective symptom recall. PMID- 15901522 TI - Can continuous glucose monitoring provide objective documentation of hypoglycemia unawareness? AB - OBJECTIVE: To establish criteria defining hypoglycemia as detected by the continuous glucose monitoring system (CGMS) in patients with type 1 diabetes that best predict hypoglycemia unawareness (HUN), established by a validated questionnaire. METHODS: Adult patients were selected for inclusion in this study if they had long-standing type 1 diabetes, a fasting level of C peptide of < or = 0.6 ng/mL, commitment to achieving glycemic control, and a hemoglobin A1c value no higher than 9%. After clinical data and self-monitoring of plasma glucose data were collected, patients underwent a 72-hour glucose monitoring session with use of a Medtronic-MiniMed CGMS. The presence of HUN was determined by a questionnaire. Factors independently associated with HUN were estimated by multivariate independent analysis. RESULTS: Our study group consisted of 60 patients (33 women and 27 men) who ranged in age from 18 to 84 years (mean, 50.4) and had had diabetes for 5 to 56 years (mean, 23.8). The best predictor of HUN was the maximal duration of hypoglycemia, as determined by the CGMS (P = 0.001). Detection of hypoglycemic episodes with a duration of more than 90 minutes identified patients who had HUN with an 88% specificity and 75% sensitivity. HUN was also significantly associated with use of angiotensin-converting enzyme inhibitors or angiotensin receptor blockers (P = 0.003) and with a longer duration of diabetes (P = 0.008). CONCLUSION: The CGMS can be used for objective detection of patients with HUN. PMID- 15901523 TI - Hypoglycemia in hospitalized adult patients without diabetes. AB - OBJECTIVE: To study hypoglycemia in hospitalized patients without diabetes and to assess the role of underlying illness in th development of hypoglycemia and its association with outcome. METHODS: We undertook a retrospective review of medical records at a university-affiliated tertiary care teaching hospital to identify adult patients without diabetes, admitted between 1994 and 2002, who presented with a blood glucose level of < or =50 mg/dL (< or =2.8 mmol/L). RESULTS: We identified 88 patients (51 men and 37 women) who fulfilled the inclusion criteria. The mean age of the study group was 64 years. Specific contributing comorbid illnesses that could cause hypoglycemia were found in 63 patients (72%), 24 (38%) of whom had more than one comorbid condition. Chronic renal failure was found in 22 patients (25%); alcohol intoxication was present in 13 patients (15%); and liver failure, sepsis, cancer, and endocrine disorders each accounted for 11 cases of hypoglycemia (12%). Of the 88 patients, 14 (16%) died. These patients were significantly older than those who survived: mean age, 75.4 versus 61.8 years, respectively (P = 0.034). Only 2 of 51 patients with a single episode of hypoglycemia died, in comparison with 12 of 37 patients with more than one hypoglycemic episode (P = 0.001). CONCLUSION: Hypoglycemia in patients without diabetes is associated with a high mortality rate. This study of hospitalized adult patients without diabetes who had low blood glucose levels extends the observation that hypoglycemia is associated with major comorbid conditions. Thus, hypoglycemia is a marker for poor outcome and a signal to the clinician to search for underlying illnesses. PMID- 15901524 TI - Insulin autoimmunity and hypoglycemia in seven white patients. AB - OBJECTIVE: To report the clinical, biochemical, and immunologic characteristics of 7 white patients with the rare disorder of hyperinsulinemic hypoglycemia in association with spontaneously generated high titers of antibodies to human insulin. METHODS: We reviewed the clinical data, history, and symptoms of the 7 study patients and summarized the biochemical findings during a spontaneous episode of hypoglycemia. Insulin antibody binding was measured in all patients, and antibody affinity, capacity, and clonality were analyzed in 4. A mixed meal study was conducted in 2 patients. A potential mechanism for postprandial hypoglycemia is presented. RESULTS: In all 7 patients (6 women and 1 man), symptoms were neuroglycopenic, occurring primarily postprandially but during fasting in some patients. During hypoglycemia, concentrations of insulin, proinsulin, and, in most patients, C peptide considerably exceeded those observed in patients with insulinoma. These concentrations were spuriously elevated as a result of interference by the autoantibodies in the immunoassays. No patient had evidence of an insulinoma on various radiologic localization procedures directed at the pancreas. Insulin antibodies showed a high percentage of binding to human insulin--50 to 90%. Heterogeneity of antibodies regarding clonality and antibody binding sites was noted; some patients had polyclonal and some had monoclonal IgG class antibodies. Most patients had two categories of binding sites: high affinity/low capacity and low capacity/high affinity. Although the mechanism for postprandial hypoglycemia remains conjectural, prolonged elevations of postprandial concentrations of total and free insulin are consistent with the putative mechanism of a buffering effect of insulin antibodies. CONCLUSION: Insulin autoimmune hypoglycemia, although rare in any racial group and especially in white subjects, can be readily detected by high titers of insulin antibodies. Such a determination should be done in all patients undergoing evaluation for hypoglycemia. PMID- 15901525 TI - Hypertension in a patient with aldosterone deficiency. AB - OBJECTIVE: To describe a patient with aldosterone synthase deficiency, who presented with failure to thrive, hypovolemic hyponatremia, and the unexpected finding of hypertension. METHODS: We present a case report, review the related literature, and outline a possible mechanism for the concomitant occurrence of high blood pressure and hyponatremia in this patient. RESULTS: A 5-month-old infant with unambiguous female genitalia was admitted to our hospital with failure to thrive and hyponatremia. Her blood pressure was 115/88 mm Hg (>95% for age). The serum sodium concentration was 123 mEq/L (normal for age, >130), and the potassium level was 5.3 mEq/L (normal, 3.5 to 5.3). A direct renin measurement by immunochemiluminescence assay was 11,400 microU/mL (normal, <5), and the aldosterone level was 4 ng/dL (normal, 2 to 70). These findings indicated a diagnosis of aldosterone synthase deficiency. Treatment with fludrocortisone and sodium chloride was begun, but the hypertension worsened. Therapy with an angiotensin-converting enzyme inhibitor was transiently required. CONCLUSION: Angiotensin II, a potent vasoconstrictor, is an intermediate in the renin angiotensin system. We believe that this protein was the cause of the hypertension in the setting of aldosterone deficiency in our patient. PMID- 15901526 TI - Persistent tumor-induced osteomalacia confirmed by elevated postoperative levels of serum fibroblast growth factor-23 and 5-year follow-up of bone density changes. AB - OBJECTIVE: To describe a case of persistent tumor-induced osteomalacia, determine whether serum fibroblast growth factor-23 (FGF-23) levels postoperatively indicate incomplete tumor resection, and report lumbar spine and forearm bone mineral density (BMD) changes during 5 years of follow-up. METHODS: We present clinical, radiologic, histologic, and bone densitometry data as well as serum FGF 23 levels (determined with use of a novel C-terminal enzyme-linked immunosorbent assay) from the study patient and discuss these findings in the context of previous literature. RESULTS: A 52-year-old man, who presented with muscle weakness and multiple fractures, was found to have low values for serum phosphorus, serum 1,25-dihydroxyvitamin D, and maximal tubular reabsorption of phosphate per glomerular filtration rate, a high level of serum alkaline phosphatase, and a normal serum concentration of parathyroid hormone, characteristic of tumor-induced osteomalacia. Magnetic resonance imaging to evaluate an abnormality of the left foot revealed a soft tissue mass, biopsy of which confirmed the presence of a benign, phosphaturic, mesenchymal tumor. The baseline serum FGF-23 level (2,050 RU/mL) was more than 17 times the upper limit of normal for adults (23 to 118 RU/mL) and decreased substantially within 1 day after partial resection of the tumor but remained above normal postoperatively. BMD changes indicated rapid substantial recovery of vertebral BMD but ongoing loss of forearm bone density. CONCLUSION: The serum FGF-23 level is high in a substantial proportion of patients with tumor-induced osteomalacia. The postoperative above normal levels of serum FGF-23 correlated with known persistence of tumor in our study patient. In a patient with normal renal function, such as our study patient, levels of serum FGF-23 studied with use of the C-terminal enzyme-linked immunosorbent assay reached their nadir within 24 hours postoperatively. This result suggests that this assay can provide clinicians with rapid prognostic information in patients with known or suspected residual tumor. BMD should be assessed at both appendicular and axial sites in patients with persistent tumor-induced osteomalacia. PMID- 15901527 TI - Rapid progression from subclinical to symptomatic overt hypothyroidism. AB - OBJECTIVE: To report a case of Hashimoto's thyroiditis with rapid progression from subclinical to overt symptomatic hypothyroidism and to discuss the potential precipitating factors and the implications on clinical decisions about monitoring and treatment of early thyroid failure. METHODS: We describe a patient with long standing subclinical hypothyroidism who had progression to severe overt hypothyroidism during a 2-month period, without an identifiable precipitating factor. All medical care was provided at a single institution, and all relevant medical records were reviewed. RESULTS: For at least 2 years, an 84-year-old man had a pattern of subclinical hypothyroidism, including normal levels of serum free thyroxine, serum thyrotropin concentrations ranging from 4.4 to 9.6 microIU/mL, and elevated levels of anti-thyroid peroxidase antibodies. During a 2 month period, symptoms of cold intolerance, a 4.5-kg weight gain, and fatigue developed, and the patient was found to have low free thyroxine and free triiodothyronine concentrations and a serum thyrotropin concentration of 80.9 microIU/mL. The patient did not use any medication previously identified as a trigger to the development of hypothyroidism, had no exposure to iodine or contrast administration, and reported no intercurrent infection that might explain the rapid progression of hypothyroidism. CONCLUSION: Most patients with subclinical hypothyroidism have progression to overt hypothyroidism at a slow rate. Elderly patients with high antithyroid antibody titers may have a higher than previously recognized risk of rapid development of overt hypothyroidism, and earlier intervention with levothyroxine treatment may be indicated. PMID- 15901528 TI - Endocrine disturbances in empty sella syndrome: case reports and review of literature. AB - OBJECTIVE: To report 5 cases of empty sella syndrome (ESS) manifesting with various degrees of pituitary dysfunction. METHODS: We describe the initial manifestations in 5 patients with primary ESS and in previous cases of ESS reported in the English language literature. RESULTS: Review of our recent medical records identified 5 patients referred for evaluation of pituitary deficiencies in whom ESS was diagnosed. Glucocorticoid replacement was required in 3 patients, 2 of whom presented initially with symptoms of severe glucocorticoid deficiency. In each case, magnetic resonance imaging of the brain demonstrated an empty sella. CONCLUSION: Our cases suggest that endocrine abnormalities are not rare as the initial manifestation of ESS and that, contrary to many studies in the literature, the endocrine abnormalities may be quite severe. PMID- 15901529 TI - Life-threatening thyrotoxicosis induced by combination therapy with PEG interferon and ribavirin in chronic hepatitis C. PMID- 15901530 TI - Idle thoughts from an addled mind. At a sixth-grade reading level. PMID- 15901531 TI - Visual vignette. PMID- 15901535 TI - Catalysis: principles, progress, prospects. AB - In this introductory paper, we endeavour to bridge the gaps that currently exist between the three main subdivisions of catalysis: enzymatic, homogeneous and heterogeneous. Hitherto, there has been a tendency for each of these three divisions to grow separately using their own concepts, phrases and techniques. However, there is much that unites them, not least the notion of the catalytically active site and, in particular, its often unusual (constrained) state of electronic or atomic environmental disposition. We identify many points of similarity between, for example, the mode of action of, metalloenzymes on the one hand and the recent generation of transition metal ions embedded within nanoporous (usually siliceous) solids on the other. Useful unifying principles emerge from considerations of free-energy/reaction-coordinate plots. We present a number of tabulations and comparisons designed to facilitate the understanding of the mode of operation of existing, and the performance of new, catalysts. In doing so, we have drawn on our own work as well as that of others, including contributions that are to be found in this volume, with the intention of covering the great variety of catalytic phenomena. PMID- 15901536 TI - Intermediates in P450 catalysis. AB - Cytochromes P450 catalyse the insertion of one O2-derived oxygen atom in unactivated C-H bonds, and as such, are potent oxidants. A significant amount is known about the P450 catalytic cycle owing partly to the single heme group at the active site that provides spectroscopic handles in tracking various intermediates. A sophisticated array of electron paramagnetic, electron double nuclear resonance, and more traditional absorption spectroscopies have been able to identify key intermediates, while crystallography has defined the structure of the substrate-free, -bound, and oxy-complexes. What has remained elusive is the Fe(IV)=O intermediate, thought to be the active hydroxylating agent. Here, theory and especially density functional calculations have provided valuable insights. PMID- 15901537 TI - Oxidation by 2-oxoglutarate oxygenases: non-haem iron systems in catalysis and signalling. AB - The 2-oxoglutarate (2OG) and ferrous iron dependent oxygenases are a superfamily of enzymes that catalyse a wide range of reactions including hydroxylations, desaturations and oxidative ring closures. Recently, it has been discovered that they act as sensors in the hypoxic response in humans and other animals. Substrate oxidation is coupled to conversion of 2OG to succinate and carbon dioxide. Kinetic, spectroscopic and structural studies are consistent with a consensus mechanism in which ordered binding of (co)substrates enables control of reactive intermediates. Binding of the substrate to the active site triggers the enzyme for ligation of dioxygen to the metal. Oxidative decarboxylation of 2OG then generates the ferryl species thought to mediate substrate oxidation. Structural studies reveal a conserved double-stranded beta-helix core responsible for binding the iron, via a 2His-1carboxylate motif and the 2OG side chain. The rigidity of this core contrasts with the conformational flexibility of surrounding regions that are involved in binding the substrate. Here we discuss the roles of 2OG oxygenases in terms of the generic structural and mechanistic features that render the 2OG oxygenases suited for their functions. PMID- 15901538 TI - Activation of oxygen at metal surfaces. AB - Oxygen chemisorption at metal surfaces has been shown through a combination of scanning tunnelling microscopy and photoelectron spectroscopy to involve transient states that provide low energy pathways for a wide range of surface reactions including the catalytic oxidation of ammonia and hydrocarbons. The kinetically 'hot' transients are disordered and mobile, become unreactive when they form ordered structures, and are characterized by non-classical kinetic behaviour. The role of surface additives (caesium) in controlling oxygen structures and the implications of oxygen transients for theory and reaction mechanisms in applied catalysis are considered. PMID- 15901539 TI - Methods and models for studying mechanisms of redox-active enzymes. AB - Methods and models used in recent quantum chemical studies of redox-active enzymes are described. Only density functional methods are able to treat these systems with sufficient accuracy. For the most accurate of these methods, the so called hybrid methods, a fraction of exact exchange is included and the deviation from experimental energies is usually not higher than 5 kcal mol-1. The size of the models depends on the problem studied, but is usually in the range 60-100 atoms. To keep the optimized structures sufficiently close to the experimental ones, one atom in each amino acid residue is kept frozen to its position in the X ray structure. Examples from different recent studies are given with emphasis on dioxygen evolution in photosystem II and proton pumping in cytochrome oxidase. The main advantage of using theoretical methods to study these systems is that short-lived intermediates and transition states can be investigated as easily as stable structures. PMID- 15901540 TI - Hydroxylation of C-H bonds at carboxylate-bridged diiron centres. AB - Nature uses carboxylate-bridged diiron centres at the active sites of enzymes that catalyse the selective hydroxylation of hydrocarbons to alcohols. The resting diiron(III) state of the hydroxylase component of soluble methane monooxygenase enzyme is converted by two-electron transfer from an NADH-requiring reductase into the active diiron(II) form, which subsequently reacts with O2 to generate a high-valent diiron(IV) oxo species (Q) that converts CH4 into CH3OH. In this step, C-H bond activation is achieved through a transition state having a linear C...H...O unit involving a bound methyl radical. Kinetic studies of the reaction of Q with substrates CH3X, where X=H, D, CH3, NO2, CN or OH, reveal two classes of reactivity depending upon whether binding to the enzyme or C-H bond activation is rate-limiting. Access of substrates to the carboxylate-bridged diiron active site in the hydroxylase (MMOH) occurs through a series of hydrophobic pockets. In the hydroxylase component of the closely related enzyme toluene/o-xylene monooxygenase (ToMOH), substrates enter through a wide channel in the alpha-subunit of the protein that tracks a course identical to that found in the structurally homologous MMOH. Synthetic models for the carboxylate-bridged diiron centres in MMOH and ToMOH have been prepared that reproduce the stoichiometry and key geometric and physical properties of the reduced and oxidized forms of the proteins. Reactions of the diiron(II) model complexes with dioxygen similarly generate reactive intermediates, including high-valent species capable not only of hydroxylating pendant C-H bonds but also of oxidizing phosphine and sulphide groups. PMID- 15901541 TI - Active sites and states in the heterogeneous catalysis of carbon-hydrogen bonds. AB - C-H bond activation for several alkenes (ethylene, propylene, isobutene, cyclohexene and 1-hexene) and alkanes (methane, ethane, n-hexane, 2-methylpentane and 3-methylpentane) has been studied on the (111) crystal face of platinum as a function of temperature at low (10(-6) Torr) and high (>/=1 Torr) pressures in the absence and presence of hydrogen pressures (>/=10 Torr). Sum frequency generation (SFG) vibrational spectroscopy has been used to characterize the adsorbate structures and high pressure scanning tunnelling microscopy (HP-STM) has been used to monitor their surface mobility under reaction conditions during hydrogenation, dehydrogenation and CO poisoning. C-H bond dissociation occurs at low temperatures, approximately 250 K, for all of these molecules, although only at high pressures for the weakly bound alkanes because of their low desorption temperatures. Bond dissociation is known to be surface structure sensitive and we find that it is also accompanied by the restructuring of the metal surface. The presence of hydrogen slows down dehydrogenation and for some of the molecules it influences the molecular rearrangement, thus altering reaction selectivity. Surface mobility of adsorbates is essential to produce catalytic activity. When surface diffusion is inhibited by CO adsorption, ordered surface structures form and the reaction is poisoned. Ethylene hydrogenation is surface structure insensitive, while cyclohexene hydrogenation and dehydrogenation are structure sensitive. n-Hexane and other C6 alkanes form either upright or flat-lying molecules on the platinum surface which react to produce branched isomers or benzene, respectively. PMID- 15901542 TI - Metal-ligand bifunctional catalysis for asymmetric hydrogenation. AB - Chiral diphosphine/1,2-diamine-Ru(II) complexes catalyse the rapid, productive and enantioselective hydrogenation of simple ketones. The carbonyl-selective hydrogenation takes place via a non-classical metal-ligand bifunctional mechanism. The reduction of the C=O function occurs in the outer coordination sphere of an 18e trans-RuH2(diphosphine)(diamine) complex without interaction between the unsaturated moiety and the metallic centre. The Ru atom donates a hydride and the NH2 ligand delivers a proton through a pericyclic six-membered transition state, directly giving an alcoholic product without metal alkoxide formation. The enantiofaces of prochiral ketones are differentiated on the chiral molecular surface of the saturated RuH2 species. This asymmetric catalysis manifests the significance of 'kinetic' supramolecular chemistry. PMID- 15901543 TI - Computational approaches to the determination of active site structures and reaction mechanisms in heterogeneous catalysts. AB - We apply quantum chemical methods to the study of active site structures and reaction mechanisms in mesoporous silica and metal oxide catalysts. Our approach is based on the use of both molecular cluster and embedded cluster (QM/MM) techniques, where the active site and molecular complex are described using density functional theory (DFT) and the embedding matrix simulated by shell model potentials. We consider three case studies: alkene epoxidation over the microporous TS-1 catalyst; methanol synthesis on ZnO and Cu/ZnO and C-H bond activation over Li-doped MgO. PMID- 15901544 TI - Activation of diatomic molecules at solid surfaces. AB - The interaction of a diatomic molecule with a well-defined single crystal surface represents the prototype of the elementary processes involved in heterogeneous catalysis, and it can be studied down to atomic length and extremely short (fs) time-scales. Our present understanding is illustrated by examples concerning the activation of hydrogen (including also the breakdown of thermal equilibrium during rapid laser-induced associative desorption), the role of steps as 'active sites' in the dissociative adsorption of NO or N2, and the mechanism and kinetics of the catalytic oxidation of CO. PMID- 15901545 TI - Catalytic reduction of dinitrogen to ammonia at well-defined single metal sites. AB - Dinitrogen (N2) is reduced to ammonia at room temperature and 1atm with molybdenum catalysts that contain tetradentate [HIPTN3N]3- triamidoamine ligands {[HIPTN3N]3-=[{3,5-(2,4,6-i-Pr3C6H2)2C6H3NCH2CH2}3N]3-, an example being [HIPTN3N]Mo(N2)} in heptane. Slow addition of the proton source ({2,6 lutidinium}{BAr'4}; Ar'=3,5-(CF3)2C6H3) and reductant (decamethyl chromocene) assure a high yield of ammonia (63-65% in four turnovers) versus dihydrogen formation. Numerous X-ray studies, along with isolation and characterization of seven intermediates in the proposed catalytic reaction (under noncatalytic conditions), suggest that N2 is being reduced at a sterically protected, single Mo centre that cycles between states Mo(III), Mo(IV), Mo(V) and Mo(VI). PMID- 15901546 TI - Structural basis of biological nitrogen fixation. AB - Biological nitrogen fixation is mediated by the nitrogenase enzyme system that catalyses the ATP dependent reduction of atmospheric dinitrogen to ammonia. Nitrogenase consists of two component metalloproteins, the MoFe-protein with the FeMo-cofactor that provides the active site for substrate reduction, and the Fe protein that couples ATP hydrolysis to electron transfer. An overview of the nitrogenase system is presented that emphasizes the structural organization of the proteins and associated metalloclusters that have the remarkable ability to catalyse nitrogen fixation under ambient conditions. Although the mechanism of ammonia formation by nitrogenase remains enigmatic, mechanistic inferences motivated by recent developments in the areas of nitrogenase biochemistry, spectroscopy, model chemistry and computational studies are discussed within this structural framework. PMID- 15901547 TI - Commentary on industrial processes. AB - A brief commentary on the three papers presented in the session on 'industrial processes' is presented. In particular, emphasis is placed on the factors considered to be crucial for an industrial process, namely: catalyst activity, the selectivity of the overall catalysed reaction and the lifetime over which the performance is observed. Control of selectivity is viewed as being most important, and examples of how this is achieved through the use of membrane reactors and catalyst design are described. PMID- 15901548 TI - Membrane reactor technology for C5/C6 hydroisomerization. AB - In this paper, we propose an improved hydroisomerization process, making use of membrane reactor technology. Linear alkanes are selectively supplied from a hydrocarbon feed (consisting of branched and linear alkanes) through an inert tubular membrane to a packed bed of catalyst. The results indicate that n-, mono- and di-branched components in a gas mixture can be separated with a selectivity factor of greater than 20 with a zeolite membrane under dedicated parameter settings. The RON-value of the product was calculated to be as high as 90 in a single pass reactor, which is 50 points higher than the feed value. The flux through the membrane could be optimized to give a STY/ATY ratio for the reactor of 877 m-1, which falls within the limits of technical feasibility. PMID- 15901549 TI - Novel, benign, solid catalysts for the oxidation of hydrocarbons. AB - The catalytic properties of two classes of solid catalysts for the oxidation of hydrocarbons in the liquid phase are discussed: (i) microporous solids, encapsulating transition metal complexes in their cavities and (ii) titanosilicate molecular sieves. Copper acetate dimers encapsulated in molecular sieves Y, MCM-22 and VPI-5 use dioxygen to regioselectively ortho-hydroxylate L tyrosine to L-dopa, phenol to catechol and cresols to the corresponding o dihydroxy and o-quinone compounds. Monomeric copper phthalocyanine and salen complexes entrapped in zeolite-Y oxidize methane to methanol, toluene to cresols, naphthalene to naphthols, xylene to xylenols and phenol to diphenols. Trimeric mu3-oxo-bridged Co/Mn cluster complexes, encapsulated inside Y-zeolite, oxidize para-xylene, almost quantitatively, to terephthalic acid. In almost all cases, the intrinsic catalytic activity (turnover frequency) of the metal complex is enhanced very significantly, upon encapsulation in the porous solids. Spectroscopic and electrochemical studies suggest that the geometric distortions of the complex on encapsulation change the electron density at the metal ion site and its redox behaviour, thereby influencing its catalytic activity and selectivity in oxidation reactions. Titanosilicate molecular sieves can oxidize hydrocarbons using dioxygen when loaded with transition metals like Pd, Au or Ag. The structure of surface Ti ions and the type of oxo-Ti species generated on contact with oxidants depend on several factors including the method of zeolite synthesis, zeolite structure, solvent, temperature and oxidant. Although, similar oxo-Ti species are present on all the titanosilicates, their relative concentrations vary among different structures and determine the product selectivity. PMID- 15901550 TI - Controlling automotive exhaust emissions: successes and underlying science. AB - Photochemical reactions of vehicle exhaust pollutants were responsible for photochemical smog in many cities during the 1960s and 1970s. Engine improvements helped, but additional measures were needed to achieve legislated emissions levels. First oxidation catalysts lowered hydrocarbon and carbon monoxide, and later nitrogen oxides were reduced to nitrogen in a two-stage process. By the 1980s, exhaust gas could be kept stoichiometric and hydrocarbons, carbon monoxide and nitrogen oxides were simultaneously converted over a single 'three-way catalyst'. Today, advanced three-way catalyst systems emissions are exceptionally low. NOx control from lean-burn engines demands an additional approach because NO cannot be dissociated under lean conditions. Current lean-burn gasoline engine NOx control involves forming a nitrate phase and periodically enriching the exhaust to reduce it to nitrogen, and this is being modified for use on diesel engines. Selective catalytic reduction with ammonia is an alternative that can be very efficient, but it requires ammonia or a compound from which it can be obtained. Diesel engines produce particulate matter, and, because of health concerns, filtration processes are being introduced to control these emissions. On heavy duty diesel engines the exhaust gas temperature is high enough for NO in the exhaust to be oxidised over a catalyst to NO2 that smoothly oxidises particulate material (PM) in the filter. Passenger cars operate at lower temperatures, and it is necessary to periodically burn the PM in air at high temperatures. PMID- 15901552 TI - [Isolation and identification of a gene: attractin related to inhibiting erythroid differentiation in K562 cells]. AB - Differential display PCR method was used to isolate four differential display fragments between the wild type K562 cell line and the variant type K562 cell line that expressed the human beta-globin gene. After sequencing, the most remarkable different fragment,named dd1,was selected for further study. The analysis of RT-PCR and Northern blot hybridization showed that dd1 was exactly the differentiation fragment between the two cell lines. The homology analysis indicated that dd1 was matched to Attractin (GeneBank registration No.AF106861). It might be an adhesion receptor related to inhibiting erythroid differentiation based on its structure. PMID- 15901553 TI - [A study of insertion/delation polymorphism of the angiotensin-converting enzyme gene in pilots]. AB - In order to understand insertion/delation (I/D) polymorphism of the angiotensin converting enzyme (ACE) gene in pilots,and to explore the relationship between ACE gene I/D polymorphism and the performance of the pilots, the polymerase chain reaction (PCR) was used to determine the genotypes for an I/D polymorphism in intron 16 of the ACE gene in 118 pilots and 96 healthy subjects as controls. The result showed that the I/D polymorphism in intron 16 of the ACE gene was categorized into three genotypes: two deletion alleles (genotype DD), heterozygous alleles (genotype ID), and two insertion alleles (genotype II). The genotype II and I allele frequency were significantly higher in pilots (44.07% and 0.65) than that in healthy subjects (31.25% and 0.52). It is suggested that I gene of ACE may play a role in performance of the pilots. PMID- 15901554 TI - [Studies of the clone and cell expression of human lactoferritin gene]. AB - In this paper,we directly cloned 2.366 Kb cDNA sequence of human lactoferrin gene and 800 bp 5'flank regulatory sequence of beta/lactoglobulin gene from goat by PCR,then connected them with the expression vector pLNCX. The recombinant plasmid pLNCXHLF containing human lactoferrin gene cDNA was transfected into mice mammary tumor cell line MA/782 after liposome transinfection. Positive single clone cells were selected with G418 and by PCR. After proliferating,the transfected cells immobilized and cultured in sodium alginate were induced by hormone. The result of Western blotting analysis on cultured cell supernatant shows that transfected cells can express the exogenic gene and secrete hLF protein, whose MW is 34 KD. The highest amount detected by ELISA reached 65 mg/l medium/10(5) cells. The result of antibacterial experiment indicates that the recombinant hLF protein has the effect of inhibiting E.coli proliferation;moreover,its activity is superior to the commercial available hLF's. PMID- 15901555 TI - [A study of forensic individual identification used STR locus with silver staining and multiplex PCR methods]. AB - Amplification of short tandem repeat(STR) loci has become a useful tool for human identification applications. To improve throughput and efficiency for the forensic materials and gain foure and six STR locis multiplex methods with silver staining, CSF1PO,TPOX,THO1 and vWA(referred to as multiplex A), D18S51, D7S820, D13S317, D5S818, D3S1358 and Amelogenin(referred to as multiplex B) have been evaluated for use in a rape case. The products of multiplex amplication were separated in a denaturing polyacrylamide gel and analyzed with silver staining. Two multiplex amplications used in this case could provide a power of discrimination of approximately 2.43 x 10(-19). Silver staining was shown to be a validation methods for analysing the products of four and six multiplex amplications. PMID- 15901556 TI - [Analysis of GUCA1B,GNGT1 and RGS9 genes in patients with retinitis pigmentosa]. AB - To screen possible disease-causing mutations in the GUCA1B gene, GNGT1 gene,and the alternative-splicing region of RGS9 gene in 120 probands with retinitis pigmentosa,genomic DNA was collected from 120 probands with retinitis pigmentosa out of 120 families. The coding sequences of the GUCA1B and GNGT1 genes and the alternative splicing region of the RGS9 gene were analyzed by using PCR heteroduplex-SSCP method. Mutation was confirmed by DNA sequencing. A T/C polymorphism was identified in exon 1 of the GUCA1B gene in 31 of the 120 probands. Heteroduplex-SSCP analysis of the GUCA1B and GNGT1 coding regions and RGS9 alternative splicing region showed no mutations in 120 patients with retinitis pigmentosa. We found no evidence that mutation in GUCA1B,GNGT1,or RGS9 gene is a cause of retinitis pigmentosa. PMID- 15901557 TI - [Discovery of Cervus nippon Temminck-derived sentrin/SUMO]. AB - In order to identify unknown encoding cDNAs of Cervus nioppon Temminck (sika deer), we constructed a cDNA library of uterus from Jilin-Shuangyang Cervus nippon Temminck using PCR cDNA library kit. PCR products of the library were cloned into pGEM-Teasy vectors and the cDNAs were sequenced and analyzed by nucleotide homology comparison against GenBank Database using the BLAST network service. The results showed that the cDNA library contained cDNA fragments of different lengths and a full length encoding cDNA highly homologous to human sentrin-1/SUMO-1 (small ubiquitin-related modifier 1) was identified. The cDNA was deposited in GenBank under the accession number AF 242526. These show that Cervus nippon Temminck-derived sentrin/SUMO gene has been discovered from PCR cDNA library of uterus from Cervus nippon Temminck. PMID- 15901558 TI - [Sex control of silkworm by the interaction between gene and environment]. AB - The silkworm embryo of the varieties, sBan, K05, Ban1, S1 and D05 was sensitive to the high temperature and low humidity incubating condition, while that of Nong4, Xinjiu and Xiankang was more endurable. By crossing the female Xinjiu with the male of sensitive varieties, such as sBaa, K05, Ban1, S1 or D05, their female progenies were sensitive to the high temperature and low humidity during incubation and could not hatch, while the males could hatch normally. Experiment showed that silkworm sex could be controlled by the interaction between gene and environment. The sensitive period was at late stage during the embryo development. The RAPD analysis showed that the similarity index of the five temperature-sensitive varieties of sBan, K05, Ban1, S1 and D05 was 63.04-76.47%. PMID- 15901559 TI - [Cloning and expression of a gene encoding shortened cecropin A-melittin hybrid in E.coli]. AB - According to the studies of David Andreu and other scientists, we designed and synthesized a gene encoding shortened cecropin A-melittin hybrid. This hybrid consists of CA(1-7) and M(5-12). The gene was inserted into pGEMEX-1 in the site of EcoRI and BamHI. A positive clone was obtained by hybridization and DNA sequencing and was expressed in JM109(DE3). PMID- 15901560 TI - [Transfer of insecticidal protein gene from Bacillus thuringiensis into conventional maize inbred-line mediated by Agrobacterium tumefaciens]. AB - Excellent inbred-lines of maize,340 and 4112, which were used largely in hybridized combination were transformed with Agrobacterium tumefaciens. The immature embryos and their original calli were infected by A.tumefaciens LBA4404 containing plasmid pGBIL04. After 3 days of co-cultivation, the immature embryos and calli were continuously selected on the medium containing phosphinothricin (PPT) for 3 generations, then plants were regenerated. It was proved by PCR analysis that the target Bt gene had been integrated into the genome of regenerated plants. The results showed that fresh original calli from the immature embryos after pre-culture were suitable acceptors. The results also showed that it could increase the frequency of selection by properly lowering the co-culture temperature to 22 degrees . PMID- 15901561 TI - [Transformation of Populus tomentosa with anti-PLD gene]. AB - Antisense Phospholipase Dgamma( PLDgamma) gene was introduced into Populus tomentosa mediated by Agrobactrium tumefaciens. The young leaves of triploid populus were used as the material and the regeneration system of high frequency has been established. We have developed the traditional transgene method by Agrobactrium trmefaciens and obtained many transgenic plants of anti-PLDgamma gene. Tests showed that the transgenic plants can grow well on the culture medium with 0.7% NaCl. PMID- 15901562 TI - [The genetic diversity and homology of Anabaena azollae and its host plant (Azolla) based on rapd analysis]. AB - Symbiotic Anabeana azollae and its host plant Anabeana-free Azolla were isolated from 16 Azolla accessions representing different Azolla species or geographic origins.DNA polymorphic fragments were obtained by simultaneous RAPD amplification of both symbiont and host. The UPGMA clusters of Anabeana azollae and its host Azolla were established separately based on Dice coefficient caculation and a coordinated relationship was shown between Anabeana azollae and its Azolla host along both individual genetic divergence,but this genetic homology was reduced among different strains within Azolla species while the obvious mutants of Anabeana azollae were detected in some Azolla tested strains collected from different geographic area in the same host species. PMID- 15901563 TI - [Construction and application of an in silico elongation system of nucleic acid sequence]. AB - Normally it is difficult to obtain full-length cDNA sequence of novel genes. More and more expressed sequence tags(ESTs) have been obtained since the start-up of human genome project. Powerful system is badly needed for data mining on these EST sequences. Based on a personal computer coupled with Linux operating system and EST database, the Blast software and Phrap software were used to construct a platform for in silico elongation of ESTs in our lab. The performance was tested using 11386 EST sequences and 511 partial-length cDNA sequences. Results demonstrated that 8373 EST and 389 cDNA sequence were elongated using this system. Thus the platform seems to be a fast way for full-length cDNA sequence cloning of new genes. PMID- 15901564 TI - [The kinetic and mathematical model of PCR amplification experiment]. AB - The PCR technique has been set up for nearly twenty years and is becoming more and more ripe. But because of the multiple influencing factors and complicated reaction procedures,no mathematical method that can describe the PCR reaction has been given. On the basis of its elementary principle,we suggested a kinetic equation to describe the reaction procedure,Wamp=[Ntarg x (1+P)n1+0.5 x Cenz x U x P x Ceactive x (n-nl)-Ntarg x (1+n x P)] x Cu x M. This equation can describe correctly the accumulation rule of PCR product and thus build up the kinetic mathematical model of PCR reaction. The predicted CT value of PE 7700 by the kinetic-mathematical model was in accordance with the real value detected by the machine. This kinetic-mathematical model accompanied by proper detecting equipment and computer could make an automatic PCR instrument, which would produce much better result. A laboratory can predict the amount of PCR product by this model and provide accurate information for further handling of PCR product according to its own condition. In this model,the molecular basis that PCR reaction is doomed to change from exponential amplification to linear amplification had been clarified. PMID- 15901565 TI - [Rapid extraction of MtDNA from Cryptococcus neoformans and examined by electronic microscopy]. AB - Cryptococcus neoformans may be grown to the exponential phase,are broken by a combination of NovoZym234 and mechanical means,and mitochondrial DNA was extracted from DNaseI-treated mitochondrial preparation by differential centrifugation. Three pellets, including yeast cell, protoplasts, mitochondrial, were examined by transmission electronic microscopy. The resulting mtDNA is sufficiently pure for restriction endonucleases analysis and PCR in further studying. A rapid and effective method for the preparation of the mtDNA of C.neoformans was established. PMID- 15901566 TI - [The molecular biology of cytoplasmic male sterility in Chinese cabbage (Brassica campestris ssp.pekinensis)]. PMID- 15901567 TI - [Nonsyndromic hereditary deafness genes research progress and related databases]. AB - Deafness is the most prevalent sensory system impairment of human, and 70% of genetic deafness belongs to nonsyndromic hearing impairment. The total number of genes involved in nonsyndromic hereditary deafness has been estimated to above 100. So far, approximate 80 loci have been mapped to human chromosome, and 23 genes have been identified. In this article,these 23 genes were summarized systematically and some databases about hereditary deafness were provided for reference. PMID- 15901568 TI - [Advances in major histocompatibility complex research in poultry]. AB - As the development of poultry genome project, it has been acquired many advances in the study of poultry MHC. At present, we have achieved some MHC sequences, locations of MHC on chromosome and some MHC gene functions. This article give a detailed introduction about gene structure of poultry MHC and its role in immune reaction, relation with antibody and advances in poultry genome about MHC. With the development of related research, how to use the result of the study more efficiently become more and more important to the poultry MHC study. PMID- 15901569 TI - [Germ cells in the murine embryonic development]. AB - This paper reviewed the recent progress of the origin, migration and proliferation, sex determination, and genomic modification of murine germ cells during its embryonic development. Murine germ cells originate from primordial germ cells at about 7-7.5dpc. Then PGCs migrated into germinal ridge at about 12.5dpc during which Steel/c-kit signal pathway plays important roles and stopped division at 13.5dpc. The sex of germ cells was mainly determined by the soma microenvironment in the gonad. And there are essential genes for sperm formation on the Y chromosome. The de novo methylation of murine germ cells was much later than soma cells and was completed at about 18.5dpc. The X chromosome reactivation of female germ cells was finished at about 14.5-15.5dpc which was independent of sexual differentiation of germinal ridge. PMID- 15901570 TI - [Genes of metal-binding proteins and their application in bioremediation of heavy metals]. AB - Heavy metal pollution has become a global environmental hazard. The use of microorganisms and plants for the decontamination of heavy metals is recognized as a low lost and high efficiency method for cleaning up metal contamination. It shows that various metal-binding proteins such as metallothioneins (MTs) or phytochelatines (PCs) play an important role in defense systems and detoxification to heavy metals in organisms. Many genes of MTs and PCs have been cloned and utilized successfully in genetically modified bacteria and plants for increasing remediation capacity. These transgenic organisms have been displayed a great potential in bioremediation and phytoremediation of heavy metals. PMID- 15901571 TI - [Progress of molecular floral development research in rice]. AB - Rice (Oryza sativa L.) is not only one of the most important food crops in the world,but also a model plant for study of molecular developmental biology in monocots. In addition, the rice floral organs provide the basis for grain formation. Study of rice floral development has become a new focus of plant molecular genetics. Recently, notable progress has been made in study of gene regulation in rice floral development. In the review, genetic and molecular mechanisms of floral induction, floral meristem formation, and floral organ development in rice are summarized. PMID- 15901572 TI - [Antibody phage display technology: realities approaching to the dream]. PMID- 15901573 TI - [Expression vector used in chloroplast genetic transformation]. AB - Chloroplast genetic transformation is a new way of plant genetic engineering. This paper reviews the construction methods of expression vector used in chloroplast genetic transformation. It contains the homologous recombinant fragments, the chloroplast specific promoter and terminater, the selectable marker genes and the interest genes whose expression in chloroplast have been achieved. PMID- 15901574 TI - [The basic outline of the evolution of single cell life-form]. AB - In 1960s, kingdoms of organisms were charted generally in a five branching form. Later, the endosymbiont hypothesis for the mitochondria and the chloroplast was proposed. The life-form is divided into two forms, the prokaryotes (bacteria) and the eukaryotes. The study of the molecular biology made the progress faster. In 1980s, Woese, CR.asserted that two-domain view of life was no longer true, a three-domain construct, the Bacteria, the Archaea, and the Eukaryotes had to take its place. At first, phylogeny trees based on differences in the amino acid sequences, then among ribosomal RNAs and also nuclear gene from hundreds of microbial species were depicted and many mini phylogenetic trees grouped the species according to their differences in the sequences. It was found that they shared genes between their contemporaries and across the species barriers. At the root of the phylogeny tree, there was not a single common cell, it was replaced by a common ancestral community of primitive cells. Genes transfered rather freely as the transposons swapping between those cells. There was no last universal common ancestor of single cell that could be found in the revised Tree of Life, It was not easy to represent the genealogical patterns of thousands of different families of genes, in one systematic map, therefor there was no trunk at all. PMID- 15901575 TI - The costs and effects of introducing selectively trained radiographers to an A&E reporting service: a retrospective controlled before and after study. AB - The costs and effects of introducing selectively trained radiographers reporting accident and emergency (A&E) radiographs of the appendicular skeleton in a district general hospital were assessed using a retrospective controlled before and after design. Reference standard reports were compared with a random stratified sample of 200 A&E and 200 general practitioner (GP) reports before and after the intervention. GP reports were used as a non-intervention, non equivalent control group. An A&E specialist registrar judged whether incorrect A&E reports might have a clinically important effect on patient management. The effect of incorrect A&E reports on outcome was assessed by patient re-attendance to the hospital because of missed abnormalities. The annual, average and incremental costs of radiographers and radiologists reporting A&E radiographs were calculated and a sensitivity analysis was undertaken. The introduction of the radiographers resulted in a 1% (95% CI -7.9 to 5.9) fall in A&E radiograph reporting accuracy and 11% (95% CI -33.7 to 11.3) reduction of cases in which incorrect A&E reports might have a clinically important effect on patient management. Only two A&E reports (one before and one after the intervention) affected patient outcome in that a fracture missed at the first visit resulted in patient re-attendance to the X-ray Department. There was a saving of 361 pounds per annum to the X-ray Department. In conclusion this study provides further evidence that selectively trained radiographers can accurately report A&E plain radiographs and at no additional cost. PMID- 15901576 TI - 24-hour ambulatory electrocardiographic monitoring is unhelpful in the investigation of older persons with recurrent falls. AB - BACKGROUND: Although frequently used in the assessment of patients with falls, it is unclear whether 24-hour ambulatory electrocardiography contributes to their assessment in older persons. OBJECTIVE: To identify electrocardiographic abnormalities in patients with recurrent falls and case controls, and determine whether 24-hour ambulatory electrocardiography identifies causal arrhythmias for falls. DESIGN: Prospective case-control study. METHODS: 24-hour ambulatory electrocardiography recordings were compared for the type and prevalence of arrhythmias and symptom correlation in consecutive older subjects with recurrent falls attending the accident and emergency department and in case controls (no previous falls or syncope). ECG abnormalities were categorised as major (ventricular arrhythmia, pauses, <30 b.p.m., Mobitz II, complete heart block) and minor (multiple ectopics, paroxysmal atrial arrhythmia and other bradyarrhythmias). RESULTS: 128 fallers (76 +/-6 years) and 100 case controls (75 +/-5 years) were recruited. Co-medication and co-morbidity were similar in both groups. 49% (63) of recordings in fallers and 41% (41) of recordings in controls were abnormal. There was no difference between groups in the prevalence of major or minor abnormalities or of symptoms during recording (breathlessness, fatigue, chest pain and dizziness). Palpitations occurred in 10% of fallers and 13% of controls. One patient fell during monitoring with no associated rhythm abnormality. CONCLUSION: Multiple abnormalities are present on 24-hour ambulatory electrocardiography in older people whether or not they have experienced falls. 24-hour electrocardiography does not discriminate between fallers and non-fallers and is not helpful in the investigation of recurrent falls. PMID- 15901577 TI - Indwelling catheter use in home care: elderly, aged 65+, in 11 different countries in Europe. AB - OBJECTIVE: To describe possible differences/patterns in the use of indwelling urinary catheters (IUC) in Europe. DESIGN: A prospective, population-based, assessment study. SETTING: The target population was 4,455 (random sample of 405 from each of 11 countries) aged 65+ receiving home care. METHODS: The clients were assessed by using the Resident Assessment Instrument MDS-HC; epidemiological and medical characteristics of clients and service utilisation were recorded. MEASUREMENTS: The frequency of use of IUC related to the patients' activities of daily living (ADL) and cognitive functioning. RESULTS: The sample consisted of 4,010 informants: 74% female, with mean age 82.3 +/-7.3 years; men 80.9 +/-7.5 years and female 82.8 +/-7.3 years. A total of 216 (5.4%) clients were using IUC. In Italy 23% were using a catheter compared with 0% in The Netherlands. Catheter use was more common in men than in women (11.5% versus 3.3%). Use of IUC was significantly correlated to certain diseases and symptoms and increase in care burden and formal services. Twenty-six per cent of the informants with indwelling catheters scored three or more on a hierarchical ADL scale (0-6). The clients using IUC in the Nordic countries were less dependent on care than in the other European countries. Models built on multivariate analysis explained 37% of the use of IUC. Tradition and attitudes may explain the differences between the sites. CONCLUSIONS: Catheter use is associated with formal or family care burden. The need for nursing home placement ought to be considered in some cases. A stricter criterion for using IUC may be considered in the southern European countries. PMID- 15901578 TI - Percutaneous endoscopic gastrostomy; evidence of different prognosis in various patient subgroups. AB - BACKGROUND: As there are no prospective randomised trials about percutaneous endoscopic gastrostomy (PEG) insertion, the medical staff and caregivers encounter great difficulty in deciding when and if to perform this procedure. OBJECTIVE: To explore which variables are associated with increased mortality after PEG insertion. DESIGN: Prospective observational study. SETTING: Gastroenterological unit of a 500-bed community hospital. SUBJECTS: All patients over the age of 50 years referred for PEG insertion between January 1992 and December 2002. METHODS: Patients were studied for their indication for PEG insertion as well as their main medical problems, and demographic details and medical records were reviewed yearly until mortality. RESULTS: 674 patients were enrolled (mean age 80.1 years, 42% men). The median survival was worst in diabetic patients (128 days, P <0.05), patients referred from hospital (161 days, P <0.01) and patients over the age of 80 years with dementia (171 days, P <0.001). The best median survival was found among demented patients under the age of 80 (467 days, P <0.05) and women under the age of 80 referred from nursing homes (780 days, P <0.01). CONCLUSIONS: The outcome after PEG insertion is variable, with survival of over a year in many of the patients. These data are important for the medical staff, the patients and their caregivers when deciding about PEG placement. PMID- 15901579 TI - FDA ponders future of NSAIDs: Pfizer reluctantly withdraws Bextra. PMID- 15901580 TI - States, hospitals learn emergency-preparedness lessons in TOPOFF 3. PMID- 15901581 TI - Some firms will use pharmacists in Medicare chronic care project. PMID- 15901582 TI - Guggul for hypercholesterolemia. PMID- 15901583 TI - Transitioning into the life of a practicing pharmacist. PMID- 15901584 TI - Bevacizumab: improved survival at what cost? PMID- 15901585 TI - Researching the human genome. PMID- 15901586 TI - Battle for funding of pharmacy residency programs. PMID- 15901587 TI - Bevacizumab in combination chemotherapy for colorectal and other cancers. AB - PURPOSE: The pharmacology, pharmacokinetics, pharmacodynamics, clinical efficacy, adverse effects, and dosage and administration of bevacizumab in patients with colorectal and other cancers are reviewed. SUMMARY: Bevacizumab is a recombinant human monoclonal antibody that inhibits the biological activities of vascular endothelial growth factor (VEGF), a protein involved in the neovascularization of multiple malignant tumors. A single dose of bevacizumab 0.1-10 mg/kg yields a maximum concentration of 2.8-284 microg/mL and shows a dose-response relationship. Pre-clinical and clinical studies have shown that bevacizumab has both cytostatic and cytotoxic effects, resulting in a reduction in tumor growth and increases in median survival time and time to tumor progression. Bevacizumab is available as an intravenous agent and carries FDA-approved labeling for use in the first-line treatment of meta-static colorectal cancer in combination with fluorouracil-based chemotherapy. Bevacizumab 5 mg/kg is infused intravenously over 30-90 minutes every two weeks. No dosage reductions are required for patients with renal or hepatic dysfunction. Bevacizumab has also yielded preliminary evidence of efficacy for breast, non-small-cell lung, pancreatic, prostate, renal, and hepatic cancers, as well as for melanoma and acute myelogenous leukemia. The most frequent adverse effects are nausea, vomiting, headache, epistaxis, anorexia, stomatitis, dyspnea, and constipation. CONCLUSION: Bevacizumab combined with fluorouracil-based chemotherapy has become the standard of care for the first-line treatment of metastatic colorectal cancer and may prove useful for other tumors as well. PMID- 15901588 TI - Efficacy and safety of rosuvastatin in treatment of dyslipidemia. AB - PURPOSE: The chemistry, pharmacology, pharmacokinetics, drug interactions, clinical efficacy, adverse effects, dosage and administration, and place in therapy of rosuvastatin are reviewed. SUMMARY: Rosuvastatin, the latest statin to receive approved labeling by the Food and Drug Administration, has shown superior efficacy in lowering low-density-lipoprotein (LDL) cholesterol. At daily doses of 5-40 mg, rosuvastatin produces mean reductions in plasma LDL cholesterol of 45 63%, statistically greater than those achieved with equivalent doses of atorvastatin, simvastatin, and pravastatin. Rosuvastatin also improves triglyceride, non-high-density lipoprotein (HDL)-cholesterol, and HDL cholesterol levels to produce a more favorable lipid profile. Rosuvastatin's safety was studied in more than 10,000 patients, exceeding the number of patients evaluated before the launch of any other statin. Many of these patients took the drug for up to 96 weeks. With regard to muscle, renal, and hepatic toxicity and the withdrawal rate due to adverse events, rosuvastatin demonstrates a safety profile similar to that of the other marketed statins. Rosuvastatin undergoes only minor metabolism (10% of the administered dose) by the cytochrome P-450 2C9 isoenzyme. Significant drug interactions were reported with cyclosporine, gemfibrozil, warfarin, and antacids. Evidence suggests that rosuvastatin will be a valuable addition to the choices for treatment of patients with dyslipidemia. CONCLUSION: Rosuvastatin has greater efficacy in lowering LDL cholesterol and non-HDL cholesterol concentrations than the other statins. It has been shown to enable more patients to reach their LDL cholesterol goals than other statins and to do so with an acceptable safety profile. PMID- 15901589 TI - Effects of a medication assistance program on health outcomes in patients with type 2 diabetes mellitus. AB - PURPOSE: The effects of a clinic-based medication assistance program (MAP) on the health outcomes and medication use of patients with type 2 diabetes mellitus were studied. METHODS: In this retrospective analysis, data from the University of Arkansas for Medical Sciences pharmacy-managed MAP and outpatient pharmacy databases were collected for adult patients with type 2 diabetes mellitus who were monitored in the university's internal medicine clinic one year before and after enrollment in the MAP. Data on patient demographics, medication use, and disease indicators (glycosylated hemoglobin [HbA(1c)], high-density-lipoprotein [HDL] cholesterol, low-density-lipoprotein [LDL] cholesterol, total cholesterol, triglyceride, and blood pressure levels) were collected for the year before enrollment and for one year after enrollment. Statistical analyses were conducted using descriptive analyses, paired t tests, and the Wilcoxon signed rank test. RESULTS: Of the 401 patients enrolled in the internal medicine clinic who were enrolled in the MAP, sufficient data were available for 52 patients, of whom 73% were women, 50% were African American, and 48% were white. Their mean age was 59 years. All were self-paying customers, with 67.3% receiving Medicare benefits. Patients received more prescription medications (p < 0.001) and antihyperglycemic medications (p = 0.001) after enrollment in the program. Mean HbA(1c) and LDL cholesterol levels decreased significantly after enrollment (p < 0.001 for both). Mean HDL cholesterol levels and systolic and diastolic blood pressure measurements did not change significantly. CONCLUSION: A clinic-based MAP managing the use of pharmaceutical manufacturers' drug assistance programs increased indigent patients' access to antihyperglycemic medications and improved patients' clinical outcomes. PMID- 15901590 TI - Automatic replacement of albuterol nebulizer therapy by metered-dose inhaler and valved holding chamber. AB - PURPOSE: Evidence supporting the delivery of bronchodilators with a metered-dose inhaler and a valved holding chamber (MDI+VHC) in place of a small-volume nebulizer (SVN) is discussed, and the steps taken to accomplish such a conversion program at one institution are described. SUMMARY: Double-blind, randomized studies in patients with acute exacerbations of asthma have demonstrated that higher doses of albuterol delivered by MDI+VHC (4-10 puffs per dose) are as effective as 2.5 mg of albuterol sulfate delivered by SVN. Three double-blind studies support the conclusion that the two methods are equivalent with respect to both efficacy and adverse effects in patients with chronic obstructive pulmonary disease. MDI+VHC offers practical advantages over SVN, including the capacity for home use by the patient, portability, less setup time, and no need for daily disinfection. Pharmacists and respiratory therapists obtained approval through the pharmacy and therapeutics committee for respiratory therapists to convert orders for bronchodilators delivered by SVN to administration by MDI+VHC. The conversion policy allows physicians to override it, but none have exercised this option. On intensive care units (ICUs), the policy resulted in a 53% increase in the use of MDI+VHC during the six-month period after it went into effect. Respiratory therapists have been less thorough in implementing the policy for non-ICU patients. CONCLUSION: Delivery of bronchodilators by MDI+VHC is as effective as delivery by SVN but offers several advantages. A policy to switch patients from SVN to MDI+VHC for bronchodilator administration met with limited success. PMID- 15901591 TI - Warfarin maintenance dosages in the very elderly. AB - PURPOSE: The effect of age on maintenance dosages among elderly patients with a stable International Normalized Ratio (INR) was studied. METHODS: The records of all patients monitored by an anticoagulation clinic between October 2000 and July 2002 were randomly selected for review. Data collection continued until at least 50 patients were enrolled in each of the following age groups: <75, 75-79, 80-84, and >/=85 years. To be included in the analysis, patients had to have been >50 years of age, had to have had a targeted INR of 2-3, and had to have had two consecutive INR values within the targeted range not more than 60 days apart. RESULTS: A total of 532 patient charts were reviewed, and 253 met the criteria for inclusion in the study. The mean +/- S.D. age of the patients was 80.6 +/- 7.8 years (range, 55-98 years). Atrial fibrillation or flutter was the most common indication for treatment. The mean daily warfarin dose was significantly lower with increasing age; the mean +/- S.D. daily dose for patients younger than 75 years was 4.9 +/- 2.6 mg/day, while that for patients 85 years or older was 3.5 +/- 2.6 mg/day. CONCLUSION: The warfarin dosage for maintenance therapy in elderly patients, especially those older than 75 years, appeared to be inversely related to age. PMID- 15901592 TI - Perceived importance of pharmacy management skills. AB - PURPOSE: U.S. and Canadian health-system pharmacists' perceptions of the importance of managerial skills and self-ratings of skills were studied. METHODS: A questionnaire asking recipients to rate the importance of 61 pharmacy management skills and to rate their own skill levels was prepared. The instrument was mailed in 2000 to pharmacy managers in Canada. Participants in the Leadership in Healthcare Administration for Pharmacists conference in Phoenix, Arizona, received the survey at the end of the 2001 and 2002 conferences. Participants in the 2002 Department of Veterans Affairs pharmacists' conference in Memphis, Tennessee, received the survey eight weeks before the conference. RESULTS: The net response rates for the Canadian, Arizona, and Tennessee surveys were 52.7%, 56.9%, and 38.4%, respectively. The five skills rated most important in each of the three surveys were all practice foundation skills and tended to be required by all health care managers. Skills rated least important were also generally similar among the surveys. Only five skills demonstrated a significant mean difference in perceived importance among the surveys. In all three surveys, demonstrating ethical conduct was rated the most important skill and was judged by participants to be their greatest strength. Using an organized system for staying current with managerial literature was cited as the greatest weakness by the Tennessee sample and the second greatest weakness by the Canadian sample. CONCLUSION: Surveys in the United States and Canada found differences and similarities in pharmacy managers' opinions of the importance of managerial skills and in self-rated managerial strengths. Also identified were gaps in training. PMID- 15901593 TI - Extended stability of ascorbic acid in 5% dextrose injection and 0.9% sodium chloride injection. PMID- 15901594 TI - Improved control of medication use with an integrated bar-code-packaging and distribution system. PMID- 15901595 TI - The other side of the human genome. PMID- 15901596 TI - Integrin alpha(IIb)beta3 signals lead cofilin to accelerate platelet actin dynamics. AB - Cofilin, in its Ser3 dephosphorylated form, accelerates actin filament turnover in cells. We report here the role of cofilin in platelet actin assembly. Cofilin is primarily phosphorylated in the resting platelet as evidenced by a specific antibody directed against its Ser3 phosphorylated form. After stimulation with thrombin under nonstirring conditions, cofilin is reversibly dephosphorylated and transiently incorporates into the actin cytoskeleton. Its dephosphorylation is maximal 1-2 min after platelet stimulation, shortly after the peak of actin assembly occurs. Cofilin rephosphorylation begins 2 min after activation and exceeds resting levels by 5-10 min. Cofilin is dephosphorylated with identical kinetics but fails to become rephosphorylated when platelets are stimulated under stirring conditions. Cofilin is normally rephosphorylated when platelets are stimulated in the presence of Arg-Gly-Asp-Ser (RGDS) peptide or wortmannin to block alpha(IIb)beta3 cross-linking and signaling or in platelets isolated from a patient with Glanzmann thrombasthenia, which express only 2-3% of normal alpha(IIb)beta3 levels. Furthermore, actin assembly and Arp2/3 complex incorporation in the platelet actin cytoskeleton are decreased when alpha(IIb)beta3 is engaged. Our results suggest that cofilin is essential for actin dynamics mediated by outside-in signals in activated platelets. PMID- 15901597 TI - iNOS initiates and sustains metabolic arrest in hypoxic lung adenocarcinoma cells: mechanism of cell survival in solid tumor core. AB - Nitric oxide (NO) modulates cellular metabolism by competitively inhibiting the reduction of O2 at respiratory complex IV. The aim of this study was to determine whether this effect could enhance cell survival in the hypoxic solid tumor core by inducing a state of metabolic arrest in cancer cells. Mitochondria from human alveolar type II-like adenocarcinoma (A549) cells showed a fourfold increase in NO-sensitive 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM) fluorescence and sixfold increase in Ca2+-insensitive NO synthase (NOS) activity during equilibration from Po2s of 100-->23 mmHg, which was abolished by N(omega) nitro-L-arginine methyl ester-HCl (L-NAME) and the inducible NOS (iNOS) inhibitor, N6-(1-iminoethyl)-L-lysine dihydrochloride (L-NIL). Similarly, cytosolic and compartmented DAF-FM fluorescence increased in intact cells during a transition between ambient Po2 and 23 mmHg and was abolished by transfection with iNOS antisense oligonucleotides (AS-ODN). In parallel, mitochondrial membrane potential (deltapsi(m)), measured using 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolo-carbocyanine iodide (JC-1), decreased to a lower steady state in hypoxia without change in glycolytic rate, adenylate energy charge, or cell viability. However, L-NAME or iNOS AS-ODN treatment maintained deltapsi(m) at normoxic levels irrespective of hypoxia and caused a marked activation of glycolysis, destabilization energy charge, and cell death. Comparison with other cancer-derived (H441) or native tissue-derived (human bronchial epithelial; alveolar type II) lung epithelial cells revealed that the hypoxic suppression of deltapsi(m) was common to cells that expressed iNOS. The controlled dissipation of deltapsi(m), absence of an overt glycolytic activation, and conservation of viability suggest that A549 cells enter a state of metabolic suppression in hypoxia, which inherently depends on the activation of iNOS as Po2 falls. PMID- 15901598 TI - Expression and membrane localization of MCT isoforms along the length of the human intestine. AB - Recent studies from our laboratory and others have demonstrated the involvement of monocarboxylate transporter (MCT)1 in the luminal uptake of short-chain fatty acids (SCFAs) in the human intestine. Functional studies from our laboratory previously demonstrated kinetically distinct SCFA transporters on the apical and basolateral membranes of human colonocytes. Although apical SCFA uptake is mediated by the MCT1 isoform, the molecular identity of the basolateral membrane SCFA transporter(s) and whether this transporter is encoded by another MCT isoform is not known. The present studies were designed to assess the expression and membrane localization of different MCT isoforms in human small intestine and colon. Immunoblotting was performed with the purified apical and basolateral membranes from human intestinal mucosa obtained from organ donor intestine. Immunohistochemistry studies were done on paraffin-embedded sections of human colonic biopsy samples. Immunoblotting studies detected a protein band of approximately 39 kDa for MCT1, predominantly in the apical membranes. The relative abundance of MCT1 mRNA and protein increased along the length of the human intestine. MCT4 (54 kDa) and MCT5 (54 kDa) isoforms showed basolateral localization and were highly expressed in the distal colon. Immunohistochemical studies confirmed that human MCT1 antibody labeling was confined to the apical membranes, whereas MCT5 antibody staining was restricted to the basolateral membranes of the colonocytes. We speculate that distinct MCT isoforms may be involved in SCFA transport across the apical or basolateral membranes in polarized colonic epithelial cells. PMID- 15901599 TI - Mitochondrial network complexity and pathological decrease in complex I activity are tightly correlated in isolated human complex I deficiency. AB - Complex I (NADH:ubiquinone oxidoreductase) is the largest multisubunit assembly of the oxidative phosphorylation system, and its malfunction is associated with a wide variety of clinical syndromes ranging from highly progressive, often early lethal, encephalopathies to neurodegenerative disorders in adult life. The changes in mitochondrial structure and function that are at the basis of the clinical symptoms are poorly understood. Video-rate confocal microscopy of cells pulse-loaded with mitochondria-specific rhodamine 123 followed by automated analysis of form factor (combined measure of length and degree of branching), aspect ratio (measure of length), and number of revealed marked differences between primary cultures of skin fibroblasts from 13 patients with an isolated complex I deficiency. These differences were independent of the affected subunit, but plotting of the activity of complex I, normalized to that of complex IV, against the ratio of either form factor or aspect ratio to number revealed a linear relationship. Relatively small reductions in activity appeared to be associated with an increase in form factor and never with a decrease in number, whereas relatively large reductions occurred in association with a decrease in form factor and/or an increase in number. These results demonstrate that complex I activity and mitochondrial structure are tightly coupled in human isolated complex I deficiency. To further prove the relationship between aberrations in mitochondrial morphology and pathological condition, fibroblasts from two patients with a different mutation but a highly fragmented mitochondrial phenotype were fused. Full restoration of the mitochondrial network demonstrated that this change in mitochondrial morphology was indeed associated with human complex I deficiency. PMID- 15901600 TI - Stimulation of astrocyte Na+/H+ exchange activity in response to in vitro ischemia depends in part on activation of ERK1/2. AB - We recently reported that Na+/H+ exchanger isoform 1 (NHE1) activity in astrocytes is stimulated and leads to intracellular Na+ loading after oxygen and glucose deprivation (OGD). However, the underlying mechanisms for this stimulation of NHE1 activity and its impact on astrocyte function are unknown. In the present study, we investigated the role of the ERK1/2 pathway in NHE1 activation. NHE1 activity was elevated by approximately 75% in NHE1+/+ astrocytes after 2-h OGD and 1-h reoxygenation (REOX). The OGD/REOX-mediated stimulation of NHE1 was partially blocked by 30 microM PD-98059. Increased expression of phosphorylated ERK1/2 was detected in NHE1+/+ astrocytes after OGD/REOX. Moreover, stimulation of NHE1 activity disrupted not only Na+ but also Ca2+ homeostasis via reverse-mode operation of Na+/Ca2+ exchange. OGD/REOX led to a 103% increase in intracellular Ca2+ concentration ([Ca2+]i) in NHE1+/+ astrocytes in the presence of thapsigargin. Inhibition of NHE1 activity with the NHE1 inhibitor HOE-642 decreased OGD/REOX-induced elevation of [Ca2+]i by 73%. To further investigate changes of Ca2+ signaling, bradykinin-mediated Ca2+ release was evaluated. Bradykinin-mediated intracellular Ca2+ transient in NHE1+/+ astrocytes was increased by approximately 84% after OGD/REOX. However, in NHE1-/- astrocytes or NHE1+/+ astrocytes treated with HOE-642, the bradykinin-induced Ca2+ release was increased by only approximately 34%. Inhibition of the reverse mode of Na+/Ca2+ exchange abolished OGD/REOX-mediated Ca2+ rise. Together, our data suggest that ERK1/2 is involved in activation of NHE1 in astrocytes after in vitro ischemia. NHE1-mediated Na+ accumulation subsequently alters Ca2+ homeostasis via Na+/Ca2+ exchange. PMID- 15901601 TI - Novel lipid mediator aspirin-triggered lipoxin A4 induces heme oxygenase-1 in endothelial cells. AB - Lipoxins (LX) and aspirin-triggered LX (ATL) are eicosanoids generated during inflammation via transcellular biosynthetic routes that elicit distinct anti inflammatory and proresolution bioactions, including inhibition of leukocyte mediated injury, stimulation of macrophage clearance of apoptotic neutrophils, repression of proinflammatory cytokine production, and inhibition of cell proliferation and migration. Recently, it was reported that aspirin induces heme oxygenase-1 (HO-1) expression on endothelial cells (EC) in a COX-independent manner, what confers protection against prooxidant insults. However, the underlying mechanisms remain unclear. In this study, we investigated whether an aspirin-triggered lipoxin A(4) stable analog, 15-epi-16-(para-fluoro)-phenoxy lipoxin A(4) (ATL-1) was able to induce endothelial HO-1. Western blot analysis showed that ATL-1 increased HO-1 protein expression associated with increased mRNA levels on EC in a time- and concentration-dependent fashion. This phenomenon appears to be mediated by the activation of the G protein-coupled LXA(4) receptor because pertussis toxin and Boc-2, a receptor antagonist, significantly inhibited ATL-1-induced HO-1 expression. We demonstrate that treatment of EC with ATL-1 inhibited VCAM and E-selectin expression induced by TNF-alpha or IL-1beta. This inhibitory effect of the analog is modulated by HO-1 because it was blocked by SnPPIX, a competitive inhibitor that blocks HO-1 activity. Our results establish that ATL-1 induces HO-1 in human EC, revealing an undescribed mechanism for the anti-inflammatory activity of these lipid mediators. PMID- 15901602 TI - Differential susceptibility of subsarcolemmal and intermyofibrillar mitochondria to apoptotic stimuli. AB - Apoptosis can be evoked by reactive oxygen species (ROS)-induced mitochondrial release of the proapoptotic factors cytochrome c and apoptosis-inducing factor (AIF). Because skeletal muscle is composed of two mitochondrial subfractions that reside in distinct subcellular regions, we investigated the apoptotic susceptibility of subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondria. SS and IMF mitochondria exhibited a dose-dependent release of protein in response to H2O2 (0, 25, 50, and 100 microM). However, IMF mitochondria were more sensitive to H2O2 and released a 2.5-fold and 10-fold greater amount of cytochrome c and AIF, respectively, compared with SS mitochondria. This finding coincided with a 44% (P < 0.05) greater rate of opening (maximum rate of absorbance decrease, V(max)) of the protein release channel, the mitochondrial permeability transition pore (mtPTP), in IMF mitochondria. IMF mitochondria also exhibited a 47% (P < 0.05) and 60% (0.05 < P < 0.1) greater expression of the key mtPTP component voltage-dependent anion channel and cyclophilin D, respectively, along with a threefold greater cytochrome c content, but similar levels of AIF compared with SS mitochondria. Despite a lower susceptibility to H2O2-induced release, SS mitochondria possessed a 10-fold greater Bax-to-Bcl-2 ratio (P < 0.05), a 2.7-fold greater rate of ROS production, and an approximately twofold greater membrane potential compared with IMF mitochondria. The expression of the antioxidant enzyme Mn2+-superoxide dismutase was similar between subfractions. Thus the divergent protein composition and function of the mtPTP between SS and IMF mitochondria contributes to a differential release of cytochrome c and AIF in response to ROS. Given the relatively high proportion of IMF mitochondria within a muscle fiber, this subfraction is likely most important in inducing apoptosis when presented with apoptotic stimuli, ultimately leading to myonuclear decay and muscle fiber atrophy. PMID- 15901603 TI - Modulation of mitochondrial Ca2+ by nitric oxide in cultured bovine vascular endothelial cells. AB - In the present study, we used laser scanning confocal microscopy in combination with fluorescent indicator dyes to investigate the effects of nitric oxide (NO) produced endogenously by stimulation of the mitochondria-specific NO synthase (mtNOS) or applied exogenously through a NO donor, on mitochondrial Ca2+ uptake, membrane potential, and gating of mitochondrial permeability transition pore (PTP) in permeabilized cultured calf pulmonary artery endothelial (CPAE) cells. Higher concentrations (100-500 microM) of the NO donor spermine NONOate (Sper/NO) significantly reduced mitochondrial Ca2+ uptake and Ca2+ extrusion rates, whereas low concentrations of Sper/NO (<100 microM) had no effect on mitochondrial Ca2+ levels ([Ca2+]mt). Stimulation of mitochondrial NO production by incubating cells with 1 mM L-arginine also decreased mitochondrial Ca2+ uptake, whereas inhibition of mtNOS with 10 microM L-N(5)-(1-iminoethyl)ornithine resulted in a significant increase of [Ca2+]mt. Sper/NO application caused a dose-dependent sustained mitochondrial depolarization as revealed with the voltage-sensitive dye tetramethylrhodamine ethyl ester (TMRE). Blocking mtNOS hyperpolarized basal mitochondrial membrane potential and partially prevented Ca2+-induced decrease in TMRE fluorescence. Higher concentrations of Sper/NO (100-500 microM) induced PTP opening, whereas lower concentrations (<100 microM) had no effect. The data demonstrate that in calf pulmonary artery endothelial cells, stimulation of mitochondrial Ca2+ uptake can activate NO production in mitochondria that in turn can modulate mitochondrial Ca2+ uptake and efflux, demonstrating a negative feedback regulation. This mechanism may be particularly important to protect against mitochondrial Ca2+ overload under pathological conditions where cellular [NO] can reach very high levels. PMID- 15901604 TI - A randomized trial of strategies for assessing eligibility for long-term domiciliary oxygen therapy. AB - RATIONALE: Restricting oxygen administration to those who benefit is desirable. OBJECTIVE: To determine the impact of alternative strategies for assessing eligibility for domiciliary oxygen on funded oxygen use, quality of life, and costs. METHODS: We randomized applicants for domiciliary oxygen therapy to an assessment system that relied on data collected by oxygen providers at the time of application and judgments by Home Oxygen Program personnel (conventional assessment) or to a system of data collection by a respiratory therapist that included, in patients unstable at the time of initial assessment, a repeat assessment after 2 months of stability (alternative assessment). MEASUREMENTS AND MAIN RESULTS: A total of 276 applicants were allocated to the conventional arm and 270 to the alternative assessment. In the year after application, oxygen use was lower in the alternative arm with no between-group differences in mortality, quality of life, or resource use in the community. Although alternative assessment applicants had on average higher assessment costs by dollars Canadian 155 per applicant, these costs were more than offset by decreased Home Oxygen Program costs of dollars Canadian 596 per applicant using Canadian cost weights. The comparable U.S. dollar figures were dollars US 309 and dollars US 432, respectively, and the difference in cost between strategies was therefore smaller using U.S. cost weights. CONCLUSIONS: Reassessment of applicants for domiciliary oxygen after several months of stability identifies an appreciable portion of initially eligible patients who are no longer eligible, thus reducing program costs to public funders without adverse consequences on quality of life, mortality, or other resource use. PMID- 15901605 TI - Exhaled nitric oxide: a predictor of steroid response. AB - RATIONALE: The initial management of patients who present with persistent respiratory symptoms includes recognizing those with the potential to benefit from inhaled steroid therapy. To date, this has required undertaking a "trial of steroid" to identify responders. There is increasing evidence that steroid response is more likely in patients with eosinophilic airway inflammation, and this can be assessed indirectly using exhaled nitric oxide (FENO) measurements. OBJECTIVES: We aimed to assess the predictive accuracy of FENO to identify steroid response in 52 patients presenting with undiagnosed respiratory symptoms in a single-blind, fixed-sequence, placebo-controlled trial of inhaled fluticasone for 4 weeks. METHODS: Comparisons of predictive accuracy were made between FENO and other conventional predictors: peak flows, spirometry, bronchodilator response, and airway hyperresponsiveness measured at baseline. "Steroid response" was defined as change in symptoms, peak flows, spirometry, or airway hyperresponsiveness to adenosine based on established guidelines and recommendations. RESULTS: Steroid response was significantly greater in the highest FENO tertile (> 47 ppb) for each endpoint. This outcome was independent of the diagnostic label. The predictive values for FENO were significantly greater than for almost all other baseline predictors, with an optimum cut point of 47 ppb. CONCLUSIONS: FENO measurements greater than 47 ppb provide a means of predicting steroid response in patients with undiagnosed respiratory symptoms. Assessing airway inflammation is of more practical value than diagnostic labeling when considering the potential usefulness of inhaled antiinflammatory therapy. PMID- 15901606 TI - Efficacy of macrophage-activating lipopeptide-2 combined with interferon-gamma in a murine asthma model. AB - RATIONALE: The incidence and prevalence of allergic asthma, caused by Th2 mediated inflammation in response to environmental antigens, is increasing. Epidemiologic data suggest that a lack of Th1-inducing factors may play a pivotal role in the development of this disease. We have previously shown that dendritic cells treated with macrophage-activating lipopeptide-2 (MALP-2) combined with IFN gamma modulate the Th2 response toward Th1 in an in vitro allergy model. OBJECTIVE: To test in vivo efficacy of this regime, the effects of the substances were evaluated in a mouse model of allergic airway inflammation. METHODS: Female Balb/c mice were sensitized to ovalbumin, whereas control animals were sham sensitized with adjuvant only. After 4 weeks, MALP-2 and IFN-gamma or NaCl, respectively, were intratracheally instillated. After inhalational ovalbumin challenge, airway hyperreactivity (AHR) to inhaled methacholine was measured by head-out body plethysmography. The animals were subsequently killed to sample bronchoalveolar lavage fluid and lungs. RESULTS: Sensitized NaCl-treated mice developed marked AHR compared with sham-sensitized animals. This coincided with eosinophilia as well as the amplification of eotaxin and the Th2 cytokines interleukin (IL)-5 and IL-13 in the bronchoalveolar lavage fluid. Treatment of sensitized mice with MALP-2 and IFN-gamma significantly reduced AHR compared with the sensitized, NaCl-treated positive control. Eosinophilia as well as Th2 cytokines were reduced to the levels of unsensitized animals. In contrast, IL 12p70 and neutrophils were markedly increased by treatment with both substances. CONCLUSION: These data demonstrate the in vivo efficacy of MALP-2 and IFN-gamma to reduce allergic inflammation and AHR in allergic asthma. PMID- 15901607 TI - Differential proteolytic enzyme activity in eosinophilic and neutrophilic asthma. AB - RATIONALE: Asthma is characterized by both chronic inflammation and remodeling of the airways. Proteases are important mediators of inflammation, cytokine activation, and tissue remodeling. OBJECTIVES: This study investigated matrix metalloproteinase-9 (MMP-9) and neutrophil elastase (NE) enzyme activity in the sputum of subjects with different inflammatory phenotypes of asthma (eosinophilic, neutrophilic, and paucigranulocytic asthma) and in healthy control subjects. METHODS AND MEASUREMENTS: Nonsmoking adults with asthma and healthy control subjects underwent hypertonic saline challenge and sputum induction. Selected sputum portions were dispersed with dithiothreitol and assayed for MMP-9 and NE enzyme activity. MAIN RESULTS: Subjects with eosinophilic asthma had significantly more active MMP-9 (39 ng/ml) compared with those with neutrophilic asthma (10 ng/ml) and control subjects (2.5 ng/ml, p < 0.01). Although there were high levels of total MMP-9 in neutrophilic asthma (5,273 ng/ml), most (> 99%) was inactivated (and bound to tissue inhibitor of metalloproteinase-1). In neutrophilic asthma, more subjects had NE activity (39%) compared with both healthy control subjects (0%), subjects with eosinophilic asthma (6%), or subjects with paucigranulocytic asthma (0%, p < 0.05). There were strong and consistent positive correlations between interleukin-8, neutrophils, and proteolytic enzymes. MMP-9 was inversely correlated with NE (r = -0.93). CONCLUSIONS: Proteolytic enzyme activity in asthma is dependent on the underlying inflammatory phenotype and is differentially regulated with MMP-9 activity a feature of eosinophilic inflammation, and active NE in neutrophilic inflammation. PMID- 15901608 TI - Early signs of atherosclerosis in obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) is associated with several cardiovascular diseases. However, the mechanisms are not completely understood. Recent studies have shown that OSA is associated with multiple markers of endothelial damage. We hypothesized that OSA affects functional and structural properties of large arteries, contributing to atherosclerosis progression. METHODS AND MEASUREMENTS: Twelve healthy volunteers, 15 patients with mild to moderate OSA, and 15 with severe OSA matched for age, sex, and body mass index were studied by using (1) full standard overnight polysomnography; (2) carotid femoral pulse wave velocity with a noninvasive automatic device; and (3) a high definition echo-tracking device to measure intima-media thickness, diameter, and distensibility. All participants were free of hypertension, diabetes, and smoking, and were not on any medications. Patients with OSA were naive to treatment. MAIN RESULTS: Significant differences existed between control subjects and patients with mild to moderate and severe OSA (apnea-hypopnea index, 3.1 +/- 0.3, 16.2 +/- 1.7, and 55.7 +/- 5.9 events/hour, respectively) in pulse wave velocity (8.7 +/- 0.2, 9.2 +/- 0.2, and 10.3 +/- 0.2 m/second; p < 0.0001), intima-media thickness (604.4 +/- 25.2, 580.2 +/- 29.0, and 722.2 +/- 35.2 microm; p = 0.004), and carotid diameter (6,607.8 +/- 126.7, 7,152.3 +/- 114.4, and 7,539.9 +/- 161.2 microm; p < 0.0001). Multivariate analyses showed that the apnea-hypopnea index correlated independently with pulse wave velocity and intima media thickness variability (r = 0.61, p < 0.0001, and r = 0.44, p = 0.004, respectively), whereas minimal oxygen saturation correlated with the carotid diameter (r = -0.60, p < 0.0001). CONCLUSIONS: Middle-aged patients with OSA who are free of overt cardiovascular diseases have early signs of atherosclerosis. All vascular abnormalities correlated significantly with the severity of the OSA, which further supports the hypothesis that OSA plays an independent role in atherosclerosis progression. PMID- 15901609 TI - In vivo and in vitro studies of a novel cytokine, interleukin 4delta2, in pulmonary tuberculosis. AB - RATIONALE: Tuberculosis progresses despite potent Th1 responses. A putative explanation is the simultaneous presence of a subversive Th2 response. However, interpretation is confounded by interleukin 4delta2 (IL-4delta2), a splice variant and inhibitor of IL-4. OBJECTIVE: To study levels of mRNA encoding IL-4 and IL-4delta2, and their relationship to treatment and clinical parameters, in cells from lung lavage and blood from patients with pulmonary tuberculosis. METHODS: IL-4delta2, IFN-gamma, IL-4, and soluble CD30 (sCD30) levels were measured by polymerase chain reaction and relevant immunoassays in 29 patients and matched control subjects lacking responses to tuberculosis-specific antigens. RESULTS: mRNA levels for IL-4 and IL-4delta2 were elevated in unstimulated cells from blood and lung lavage of patients versus control subjects (p < 0.005). In control subjects, there were low basal levels of IL-4 and IL-4delta2 mRNA expressed mainly by non-T cells (p < 0.05). However, in patients, there were greater levels of mRNA for both cytokines in both T- and non-T-cell populations (p < 0.05 compared with control subjects). Radiologic disease correlated with the IL-4/IFN-gamma ratio and sCD30 (p < 0.005). After chemotherapy, IL-4 mRNA levels remained unchanged, whereas IL-4delta2 increased in parallel with IFN-gamma (p < 0.05). Sonicates of Mycobacterium tuberculosis upregulated expression of IL-4 relative to IL-4delta2 in mononuclear cell cultures from patients (p < 0.05). CONCLUSIONS: A Th2-like response, prominent in T cells and driven by tuberculosis antigen, is present in tuberculosis and modulated by treatment, suggesting a role for IL-4 and IL-4delta2 in the pathogenesis of tuberculosis and their ratio as a possible marker of disease activity. The specific antigens inducing the IL-4 response require identification to facilitate future vaccine development strategies. PMID- 15901610 TI - Inhaled corticosteroids in chronic obstructive pulmonary disease: results from two observational designs free of immortal time bias. AB - RATIONALE: Recent cohort studies in chronic obstructive pulmonary disease (COPD) have questioned the validity of previously reported associations between inhaled corticosteroids (ICS) and reductions in mortality and rehospitalization in observational studies. Using time-dependent versions of statistical survival models, these studies have suggested immortal time bias as responsible for the proposed beneficial association. OBJECTIVES: We explored the extent of this bias in a study of patients with COPD monitored for a year from COPD discharge with two designs free of any immortal time bias in the General Practice Research Database in the United Kingdom. METHODS: In Design 1, we used only patients whose treatment status was defined on the same day of discharge to obtain a matched cohort based on propensity scores, which were derived from the patient-level baseline characteristics. In Design 2, we identified all in the study cohort who experienced death or rehospitalization and then matched each case to up to four noncases by randomly sampling from the cohort risk sets without regard to treatment status. MEASUREMENTS AND MAIN RESULTS: The propensity scores matched cohort analysis of 786 patients without a wait time found a significant risk reduction associated with use of ICS: hazard ratio, 0.69 (95% confidence interval, 0.52-0.93). The matched nested case-control analysis of 2,222 patients, designed without regard to exposure status and hence free of immortal time bias, gave a similar association with exposure to ICS in the last 6-month period: hazard ratio, 0.71 (0.56-0.90). CONCLUSIONS: We conclude that immortal time bias cannot account for the risk reduction associated with ICS exposure in observational studies. PMID- 15901611 TI - Effect of prone position on regional shunt, aeration, and perfusion in experimental acute lung injury. AB - RATIONALE: The prone position is used to improve gas exchange in patients with acute respiratory distress syndrome. However, the regional mechanism by which the prone position improves gas exchange in acutely injured lungs is still incompletely defined. METHODS: We used positron emission tomography imaging of [(13)N]nitrogen to assess the regional distribution of pulmonary shunt, aeration, perfusion, and ventilation in seven surfactant-depleted sheep in supine and prone positions. RESULTS: In the supine position, the dorsal lung regions had a high shunt fraction, high perfusion, and poor aeration. The prone position was associated with an increase in lung gas content and with a more uniform distribution of aeration, as the increase in aeration in dorsal lung regions was not offset by loss of aeration in ventral regions. Consequently, the shunt fraction decreased in dorsal regions in the prone position without a concomitant impairment of gas exchange in ventral regions, thus leading to a significant increase in the fraction of pulmonary perfusion participating in gas exchange. In addition, the vertical distribution of specific alveolar ventilation became more uniform in the prone position. A biphasic relation between regional shunt fraction and gas fraction showed low shunt for values of gas fraction higher than a threshold, and a steep linear increase in shunt for lower values of gas fraction. CONCLUSION: In a surfactant-deficient model of lung injury, the prone position improved gas exchange by restoring aeration and decreasing shunt while preserving perfusion in dorsal lung regions, and by making the distribution of ventilation more uniform. PMID- 15901612 TI - Genomewide screen for pulmonary function in 200 families ascertained for asthma. AB - Changes in pulmonary function are important in determining asthma outcome. Genetic factors may influence airway obstruction in asthma. We performed a genomewide screen in 200 families of probands objectively diagnosed with asthma in the 1960s to identify chromosomal regions related to changes in pre- and postbronchodilator lung function (FEV1, VC, and FEV1%VC) and assess influences of early-life smoke exposure. Smoking (pack-years), age, sex, and height were covariates in variance component analyses. Significant evidence for linkage of pre- and postbronchodilator FEV1%VC was obtained for chromosome 2q32 (LOD,4.9, increasing to 6.03 with additional fine-mapping markers, and 3.2, respectively). Linkage existed for chromosome 5q for pre- and postbronchodilator VC (likelihood of disease [LOD], 1.8 and 2.6, respectively). Results for pre- and postbronchodilator FEV1 were less significant (LOD, 1.5 and 1.6, chromosomes 11p and 10q, respectively). Results were not affected by passive smoke exposure. There is significant evidence for linkage of FEV1%VC to chromosome 2q32 in families of probands with asthma, 35 cM proximal from linkage previously observed in families of probands with early-onset chronic obstructive pulmonary disease. Thus, there may be multiple genes on chromosome 2q that are important in determining presence and degree of airflow limitation in families ascertained for obstructive airway disease. PMID- 15901613 TI - Cerebrovascular response to carbon dioxide in patients with congestive heart failure. AB - RATIONALE: Cerebrovascular reactivity to CO(2) provides an important counterregulatory mechanism that serves to minimize the change in H(+) at the central chemoreceptor, thereby stabilizing the breathing pattern in the face of perturbations in Pa(CO(2)). However, there are no studies relating cerebral circulation abnormality to the presence or absence of central sleep apnea in patients with heart failure. OBJECTIVES: To determine whether patients with congestive heart failure and central sleep apnea have an attenuated cerebrovascular responsibility to CO(2). METHODS: Cerebral blood flow velocity in the middle cerebral artery was measured in patients with stable congestive heart failure with (n = 9) and without (n = 8) central sleep apnea using transcranial ultrasound during eucapnia (room air), hypercapnia (inspired CO(2), 3 and 5%), and hypocapnia (voluntary hyperventilation). In addition, eight subjects with apnea and nine without apnea performed a 20-second breath-hold to investigate the dynamic cerebrovascular response to apnea. MEASUREMENTS AND MAIN RESULTS: The overall cerebrovascular reactivity to CO(2) (hyper- and hypocapnia) was lower in patients with apnea than in the control group (1.8 +/- 0.2 vs. 2.5 +/- 0.2%/mm Hg, p < 0.05), mainly due to the prominent reduction of cerebrovascular reactivity to hypocapnia (1.2 +/- 0.3 vs. 2.2 +/- 0.1%/mm Hg, p < 0.05). Similarly, brain blood flow demonstrated a smaller surge after a 20-second breath hold (peak velocity, 119 +/- 4 vs. 141 +/- 8% of baseline, p < 0.05). CONCLUSION: Patients with central sleep apnea have a diminished cerebrovascular response to PET(CO(2)), especially to hypocapnia. The compromised cerebrovascular reactivity to CO(2) might affect stability of the breathing pattern by causing ventilatory overshooting during hypercapnia and undershooting during hypocapnia. PMID- 15901614 TI - Heme oxygenase-1: from bench to bedside. AB - As aspects of basic science come to play an increasingly prominent role in clinical medicine, heme oxygenase-1 is one of several molecules emerging as a central player in diseases of the lung and intensive care unit. Although the apparent raison d'etre of this enzyme is to dispose of heme, its activity results in cytoprotection against oxidative injury and cellular stresses. As the lung interfaces directly with an oxidizing environment, it is expected that heme oxygenase-1 would be involved in many aspects of lung health and disease. The protective effects of heme oxygenase-1 and products of its enzymatic activity, including carbon monoxide, biliverdin and bilirubin, and ferritin, have opened the door to potential therapeutic and disease-monitoring possibilities that one day may be applicable to pulmonary medicine. This article introduces readers to the history of heme oxygenase research, the role of this enzyme in the lung, and related new developments to look forward to in the fields of pulmonary and critical care medicine. PMID- 15901615 TI - 4E-binding protein phosphorylation and eukaryotic initiation factor-4E release are required for airway smooth muscle hypertrophy. AB - The molecular mechanisms of airway smooth muscle hypertrophy, a feature of severe asthma, are poorly understood. We previously established a conditionally immortalized human bronchial smooth muscle cell line with a temperature-sensitive SV40 large T antigen. Temperature shift and loss of large T cause G1-phase cell cycle arrest that is accompanied by increased airway smooth muscle cell size. In the present study, we hypothesized that phosphorylation of eukaryotic initiation factor-4E (eIF4E)-binding protein (4E-BP), which subsequently releases eIF4E and initiates cap-dependent mRNA translation, was required for airway smooth muscle hypertrophy. Treatment of cells with chemical inhibitors of PI 3-kinase and mammalian target of rapamycin blocked protein synthesis and cell growth while decreasing the phosphorylation of 4E-BP and increasing the binding of 4E-BP to eIF4E, consistent with the notion that 4E-BP1 phosphorylation and eIF4E function are required for hypertrophy. To test this directly, we infected cells with a retrovirus encoding a phosphorylation site mutant of 4E-BP1 (AA-4E-BP-1) that dominantly inhibits eIF4E. Upon temperature shift, cells infected with AA-4E-BP 1, but not empty vector, failed to undergo hypertrophic growth. We conclude that phosphorylation of 4E-BP, eIF4E release, and cap-dependent protein synthesis are required for hypertrophy of human airway smooth muscle cells. PMID- 15901616 TI - Interleukin-17 as a recruitment and survival factor for airway macrophages in allergic airway inflammation. AB - Recent data indicate that the proinflammatory cytokine, interleukin (IL)-17, stimulates certain effector functions of human macrophages. We evaluated whether IL-17 mediates allergen-induced accumulation of airway macrophages and, if so, whether such an effect relates to the control of macrophage recruitment and survival. BALB/c mice were sensitized and challenged with ovalbumin. Three hours before challenge an anti-mouse IL-17 mAb (a-IL-17) was administered. Sampling was conducted 24 h after the allergen challenge. In vitro chemotaxis assay for blood monocytes and culture of airway macrophages, immunocytochemistry for Fas-antigen, and matrix metalloproteinase-9 (MMP-9) were used to determine the effect of IL-17 on the recruitment, survival, and activity of airway macrophages. A-IL-17 reduced the number of airway neutrophils and macrophages after allergen challenge. In vitro, recombinant IL-17 induced migration of blood monocytes and prolonged survival of airway macrophages. A-IL-17 also increased the expression of Fas antigen in airway macrophages in vivo. Finally, the expression of MMP-9 by airway neutrophils and macrophages in vivo was downregulated by a-IL-17. This study indicates that endogenous IL-17 mediates the accumulation of macrophages during allergen-induced airway inflammation. IL-17 exerts its effects by acting directly on airway macrophages by promoting their recruitment and survival. Furthermore, IL-17 is involved in controlling the proteolytic activity of macrophages and neutrophils in allergen-induced airway inflammation. PMID- 15901617 TI - Nitric oxide, hemoglobin, and hypoxic vasodilation. PMID- 15901618 TI - A centennial history of research on asthma pathogenesis. PMID- 15901620 TI - Effectiveness of highly active antiretroviral therapy among injection drug users with late-stage human immunodeficiency virus infection. AB - Highly active antiretroviral therapy (HAART) has been shown to be effective in different populations, but data among injection drug users are limited. Human immunodeficiency virus-infected injection drug users recruited into the Acquired Immunodeficiency Syndrome Link to Intravenous Experiences (ALIVE) Study as early as 1988 were tested semiannually to identify their first CD4-positive T lymphocyte cell count below 200/microl; they were followed for mortality through 2002. Visits were categorized into the pre-HAART (before mid-1996) and the HAART eras and further categorized by HAART use. Survival analysis with staggered entry was used to evaluate the effect of HAART on acquired immunodeficiency syndrome related mortality, adjusting for other medications and demographic, clinical, and behavioral factors. Among 665 participants, 258 died during 2,402 person-years of follow-up. Compared with survival in the pre-HAART era, survival in the HAART era was shown by multivariate analysis to be improved for both those who did and did not receive HAART (relative hazards = 0.06 and 0.33, respectively; p < 0.001). Inferences were unchanged after restricting analyses to data starting with 1993 and considerations of lead-time bias and human immunodeficiency viral load. The annual CD4-positive T-lymphocyte cell decline was less in untreated HAART-era participants than in pre-HAART-era participants (-10/microl vs. -37/microl, respectively), suggesting that changing indications for treatment may have contributed to improved survival and that analyses restricted to the HAART era probably underestimate HAART effectiveness. PMID- 15901621 TI - Exposure to Chlamydia pneumoniae infection and progression of age-related macular degeneration. AB - Recent studies have found an association between exposure to Chlamydia pneumoniae infection and risk of age-related macular degeneration (AMD). To assess a potential risk of AMD progression posed by exposure to C. pneumoniae, the authors reexamined Australian residents in 2001-2002 who were aged 51-89 years with early AMD at baseline (1992-1995). Examination included macular photography and an enzyme-linked immunosorbent assay to determine antibody titers to the elementary bodies from C. pneumoniae AR39. AMD progression was assessed quantitatively, using both coarse and fine progression steps following an international classification for AMD grading, and also qualitatively, by side-by-side comparison of baseline and follow-up macular photographs. Serologic data were available for 246 of 254 (97%) subjects. AMD progression was associated with a higher antibody titer. After adjustment for age, smoking, family history of AMD, history of cardiovascular diseases, and source study, the subjects in the upper tertiles of antibody titers were 2.1 (95% confidence interval: 0.92, 4.69), 2.6 (95% confidence interval: 1.24, 5.41), and 3.0 (95% confidence interval: 1.46, 6.37) times more at risk of progression than those in the lowest tertile, using three definitions of progression, respectively. The fact that seroreactivity to C. pneumoniae was independently associated with the risk of AMD progression suggests that C. pneumoniae infection may be an additional risk factor for AMD progression. PMID- 15901622 TI - Retinal degeneration and other eye disorders in wives of farmer pesticide applicators enrolled in the agricultural health study. AB - Retinal degeneration is the leading cause of visual impairment in older adults. An association between retinal degeneration and fungicide use was observed previously among farmer pesticide applicators in the Agricultural Health Study, a large study of farm families from Iowa and North Carolina. The objective of this investigation was to determine whether wives of these farmer pesticide applicators were at increased risk of retinal degeneration. Self-reported cross sectional data obtained via questionnaire between 1993 and 1997 from 31,173 wives were used. Associations of specific pesticides and groups of pesticides based on function (fungicides, herbicides, insecticides, and fumigants) or chemical structure (organophosphates, organochlorines, and carbamates) with eye disorders were evaluated using logistic and hierarchical logistic regression analyses. Self reported retinal degeneration was associated with the wife's fungicide use (odds ratio = 1.9, 95% confidence interval: 1.2, 3.1) after adjustment for age and state of residence. Specific fungicides that appeared to drive this association were maneb or mancozeb and ziram. No associations between pesticide use and other eye disorders were found. Although these findings for retinal degeneration are based solely on self-reported disease, they are consistent with those reported for farmer pesticide applicators. These findings suggest that exposure to some fungicides and other pesticides may increase the risk of retinal degeneration and warrant further investigation. PMID- 15901623 TI - Particulate air pollution and the rate of hospitalization for congestive heart failure among medicare beneficiaries in Pittsburgh, Pennsylvania. AB - The authors used a case-crossover approach to evaluate the association between ambient air pollution and the rate of hospitalization for congestive heart failure among Medicare recipients (aged > or =65 years) residing in Allegheny County (Pittsburgh area), Pennsylvania, during 1987-1999. They also explored effect modification by age, gender, and specific secondary diagnoses. During follow-up, 55,019 patients were admitted with a primary diagnosis of congestive heart failure. In single-pollutant models, particulate matter with an aerodynamic diameter of <10 microm (PM(10)), carbon monoxide, nitrogen dioxide, and sulfur dioxide-but not ozone-were positively and significantly associated with the rate of admission on the same day. The strongest associations were observed with carbon monoxide, nitrogen dioxide, and PM(10). The associations with carbon monoxide and nitrogen dioxide were the most robust in two-pollutant models, remaining statistically significant even after adjusting for other pollutants. Patients with a recent myocardial infarction were at greater risk of particulate related admission; otherwise, there was no significant effect modification by age, gender, or other secondary diagnoses. These results suggest that short-term elevations in air pollution from traffic-related sources may trigger acute cardiac decompensation in heart failure patients and that those with certain comorbid conditions may be more susceptible to these effects. PMID- 15901624 TI - Lung cancer risk: effect of dairy farming and the consequence of removing that occupational exposure. AB - The aim of this study was to confirm the exposure-dependent reduction in lung cancer risk reported for dairy farmers exposed to endotoxin and to evaluate the consequence of leaving dairy farming and taking employment in industry or services, where exposure to microbial agents is lower. Standardized mortality ratios, with 95% confidence intervals, for 2,561 self-employed dairy farmers were estimated, considering the general population of Veneto, Italy, from 1970 to 1998 as the reference. Sixty-two lung cancer cases, whose information was checked against clinical records, were compared with 333 controls in a cohort-nested case control study. Odds ratios with 95% confidence intervals were estimated by logistic regression analysis. A downward trend of standardized mortality ratios for lung cancer across tertiles of number of dairy cattle on the farm was significant (p < 0.05) from 1970 to 1984 but not from 1985 to 1998, when most subjects were no longer dairy farmers. Age- and smoking-adjusted odds ratios for lung cancer significantly decreased with increasing number of dairy cattle (p for trend = 0.001) for workers for whom < or =15 but not >15 years had elapsed from the end of work to the end of follow-up. In conclusion, increased levels of endotoxin (or other associated environmental factors) might be protective against lung cancer; protection diminishes over time after that exposure is removed. PMID- 15901625 TI - Cancer risk in persons with oral cleft--a population-based study of 8,093 cases. AB - The authors conducted a nationwide study of the occurrence of cancer among 8,093 Danish oral cleft cases born in 1936 through 1998 and followed in the Danish Cancer Registry from 1968 through 1998, a total of 175,863 person-years, to assess a possible association between cancer and oral clefts. Observed and expected numbers of cancers among oral cleft cases were summarized as the overall and as 52 site-specific standardized incidence ratios. The expected overall number of all cancers was 131, but 140 incident cancers were found, corresponding to a standardized incidence ratio of 1.07 (95% confidence interval (CI): 0.90, 1.26). Analyses of the 52 sites for all oral cleft cases and analyses stratified for three cleft subgroups and the two sexes revealed only a few significant associations: an increased occurrence of breast cancer among females born with cleft lip and/or cleft palate (standardized incidence ratio (SIR) = 1.52, 95% CI: 1.05, 2.14), primary brain cancer among females born with cleft palate (SIR = 3.11, 95% CI: 1.14, 6.78), and primary lung cancer among males born with both cleft lip and cleft palate (SIR = 2.49, 95% CI: 1.00, 5.14). The results do not provide evidence for an increased overall cancer risk for individuals born with oral clefts. PMID- 15901626 TI - Role of goiter and of menstrual and reproductive factors in thyroid cancer: a population-based case-control study in New Caledonia (South Pacific), a very high incidence area. AB - Exceptionally high incidence rates of thyroid cancer have been reported for Melanesian women in New Caledonia (South Pacific). To investigate the occurrence of thyroid cancer in that country and to clarify the role of goiter and hormonal factors in that disease in women, a countrywide population-based case-control study was conducted in 1993-1999. The study included 293 cases, identified through pathology registers and whose thyroid cancer was verified histologically, and 354 population controls. Thyroid cancer was associated with goiter, age at menarche, irregular menstruation, and hysterectomy. There was a dose-response trend with number of full-term pregnancies (p = 0.01), with an odds ratio of 2.2 (95% confidence interval: 1.1, 4.3) for women with eight or more pregnancies. Miscarriage, particularly as an outcome of the first pregnancy, was also indicated as a risk factor. The association between voluntary abortion and thyroid microcarcinoma could be explained by enhanced medical surveillance and improved cancer detection in women undergoing abortion. Oral contraceptives and hormone replacement therapy were unrelated to thyroid cancer. The very high birth rate among Melanesian women in New Caledonia, as well as late age at menarche, may explain, in part, their elevated incidence of thyroid cancer. PMID- 15901627 TI - Incidence of primary headache: a Danish epidemiologic follow-up study. AB - The incidence of migraine in a general population has been assessed in few longitudinal studies, and the incidence of tension-type headache has never been assessed. The authors aimed to assess the incidence of migraine and tension-type headache in Denmark by conducting a 12-year follow-up study of a general population (1989-2001). The design and methods of follow-up replicated the baseline study exactly, including use of the International Headache Society's diagnostic criteria and administration of headache diagnostic interviews by a physician. Of 740 persons aged 25-64 years examined in 1989, 673 were eligible in 2001 and 549 (81.6%) participated. The incidence of migraine was 8.1 per 1,000 person-years (male:female ratio, 1:6), and the incidence of frequent tension-type headache was 14.2 per 1,000 person-years (male:female ratio, 1:3). Both rates decreased with age. The incidence of migraine was higher than that previously calculated from cross-sectional studies. Risk factors for migraine were familial disposition, no vocational education, a high work load, and frequent tension-type headache. For tension-type headache, risk factors were poor self-rated health, inability to relax after work, and sleeping few hours per night. The gender difference for tension-type headache differed from that for migraine, and no association with educational level was observed. PMID- 15901628 TI - Birth weight, infant weight gain, and cause-specific mortality: the Hertfordshire Cohort Study. AB - Low birth weight, a marker of adverse intrauterine circumstances, is known to be associated with a range of disease outcomes in later life, including coronary heart disease, hypertension, type 2 diabetes, and osteoporosis. However, it may also decrease the risk of other common conditions, most notably neoplastic disease. The authors describe the associations between birth weight, infant weight gain, and a range of mortality outcomes in the Hertfordshire Cohort. This study included 37,615 men and women born in Hertfordshire, United Kingdom, in 1911-1939; 7,916 had died by the end of 1999. For men, lower birth weight was associated with increased risk of mortality from circulatory disease (hazard ratio per standard deviation decrease in birth weight = 1.08, 95% confidence interval: 1.04, 1.11) and from accidental falls but with decreased risk of mortality from cancer (hazard ratio per standard deviation decrease in birth weight = 0.94, 95% confidence interval: 0.90, 0.98). For women, lower birth weight was associated with a significantly (p < 0.05) increased risk of mortality from circulatory and musculoskeletal disease, pneumonia, injury, and diabetes. Overall, a one-standard-deviation increase in birth weight reduced all-cause mortality risk by age 75 years by 0.86% for both men and women. PMID- 15901629 TI - The associations of physical activity and adiposity with alanine aminotransferase and gamma-glutamyltransferase. AB - The mechanisms linking obesity and inactivity to diabetes mellitus are unclear. Recent studies have shown associations of alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) with diabetes. In a random sample of 3,789 British women aged 60-79 years, the authors examined the associations of obesity and physical activity with ALT and GGT (1999-2001). Both body mass index and waist:hip ratio were independently (of each other, physical activity, alcohol consumption, smoking, and childhood and adulthood social class) positively and linearly associated with ALT and GGT. In adjusted models, a one-standard deviation increase in body mass index was associated with a 0.46-units/liter (95% confidence interval (CI): 0.16, 0.75) increase in ALT and a 2.14-units/liter (95% CI: 0.99, 3.30) increase in GGT. Similar results for a one-standard-deviation increase in waist:hip ratio were 13.96 (95% CI: 10.44, 17.48) for ALT and 39.44 (95% CI: 25.89, 52.98) for GGT. Frequency of physical activity was inversely and linearly associated with GGT in fully adjusted models, but the inverse association with ALT was attenuated towards the null after adjustment for body mass index and waist:hip ratio. Adjustment for ALT and GGT resulted in some attenuation of the strong linear associations of body mass index and waist:hip ratio with diabetes. These findings provide some support for the suggestion that the relation between obesity and diabetes is, at least in part, mediated by liver pathology. PMID- 15901630 TI - Re: "Mortality from Solid Cancers among Workers in Formaldehyde Industries". PMID- 15901632 TI - Comparison of long term outcome of patients with rheumatoid arthritis presenting with undifferentiated arthritis or with rheumatoid arthritis: an observational cohort study. AB - BACKGROUND: The outcome of undifferentiated arthritis (UA) ranges from remission to rheumatoid arthritis (RA) fulfilling the American College of Rheumatology (ACR) classification criteria. OBJECTIVES: To report the outcome of UA after 1 year of follow up and compare the disease course of patients who presented with UA, but evolved into RA within 1 year (UA-RA group), with that of patients who presented with RA fulfilling the ACR criteria (RA-RA group). METHODS: The diagnosis of 330 patients who presented with UA was recorded at 1 year. The UA-RA and RA-RA groups were then followed up for 3 more years. Outcome measurements were radiographic progression, disease activity, and functional capacity. RESULTS: From 330 patients who were diagnosed UA, 91 had evolved into RA at 1 year; 62 patients had presented with RA. No significant differences were detected between the UA-RA and RA-RA groups in median Sharp/van der Heijde score at baseline, radiographic progression rates, disease activity, and functional capacity. However, significantly more disease modifying antirheumatic drugs were prescribed in the RA-RA group. CONCLUSION: The disease outcome of patients who present with UA that evolves into RA within 1 year is the same as that of patients who present with RA as measured by radiographic progression, disease activity, and functional capacity. PMID- 15901633 TI - Inhibition of collagen gene expression in systemic sclerosis dermal fibroblasts by mithramycin. AB - BACKGROUND: The anti-tumour antibiotic mithramycin is also a potent inhibitor of fibrosis after glaucoma surgery. This drug displays high affinity binding to GC rich sequences in DNA, including those present in the promoter of the gene encoding the alpha1 chain of type I collagen (COL1A1). OBJECTIVE: To evaluate the effects of mithramycin on COL1A1 expression in systemic sclerosis fibroblasts. METHODS: Confluent cultures of dermal fibroblasts from patients with recent onset diffuse systemic sclerosis were treated with mithramycin in vitro. Cell viability and protein expression were examined by fluorescence and confocal imaging. Type I collagen production was analysed by confocal imaging and metabolic labelling. COL1A1 messenger RNA levels and stability were assessed by northern hybridisation, and COL1A1 transcription was examined by transient transfections. RESULTS: Treatment of systemic sclerosis fibroblasts with mithramycin (10-100 nmol/l) did not cause significant cytotoxicity. Type I collagen biosynthesis decreased by 33-40% and 50-70% in cells cultured with mithramycin at 10 nmol/l and 100 nmol/l, respectively. Mithramycin at 50 nmol/l decreased COL1A1 mRNA levels by 40-60%. The effects of mithramycin on collagen gene expression were mediated by transcriptional and post-transcriptional mechanisms as shown by the reduction of COL1A1 promoter activity and by a decrease in the stability of these transcripts, respectively. CONCLUSIONS: Mithramycin causes potent inhibition of collagen production and gene expression in systemic sclerosis dermal fibroblasts in vitro in the absence of cytotoxic effects. These results suggest that this drug may be an effective treatment for the fibrotic process which is the hallmark of systemic sclerosis. PMID- 15901634 TI - Double blind, randomised, placebo controlled study of leflunomide in the treatment of active ankylosing spondylitis. AB - OBJECTIVE: To assess the efficacy and safety of leflunomide in active ankylosing spondylitis (AS) compared with placebo in a 24 week pilot study. METHODS: In a single centre randomised, double blind, placebo controlled study, 45 patients with active AS were randomised to either leflunomide 20 mg daily (n=30) or placebo (n=15). Active disease was defined as a score of >or=4 on the Bath ankylosing spondylitis disease activity index (0-10), and pain of >or=4 on a visual analogue scale (0-10). The primary efficacy variable at week 24 was the 20% response rate, as recommended by the Assessments in Ankylosing Spondylitis (ASAS) working group. Secondary outcome variables included general wellbeing, metrology index, swollen joint count, erythrocyte sedimentation rate, and C reactive protein. RESULTS: In all, 13 women and 32 men were studied. Demographic and disease indices were comparable between the two treatment groups at baseline. The rate of ASAS 20% responders was not significantly different: 27% in the leflunomide treated patients and 20% in the placebo group (95% confidence interval, -32% to 19%). No significant differences were found between the treatment groups in mean changes of the secondary outcome variables. Eleven patients were withdrawn prematurely from the study because of adverse events (7), lack of efficacy (3), and non-compliance (1). Most frequently adverse events were gastrointestinal side effects and skin disorders. CONCLUSIONS: In this placebo controlled study, leflunomide treatment did not result in a significant improvement of the ASAS 20% response in active ankylosing spondylitis. No unexpected or severe adverse events occurred. PMID- 15901635 TI - Muscle strength, pain, and disease activity explain individual subdimensions of the Health Assessment Questionnaire disability index, especially in women with rheumatoid arthritis. AB - OBJECTIVE: To study the extent to which muscle strength and performance, pain, and disease activity are associated with the total Health Assessment Questionnaire (HAQ) disability index and its subdimensions in male and female patients with rheumatoid arthritis. METHODS: HAQ for functional capacity was completed by 135 patients with rheumatoid arthritis referred for orthopaedic surgery (74% women; mean (SD) age 62 (10) years; disease duration 19 (13) years, 70% positive for rheumatoid factor). Knee extension, trunk extension and flexion, grip strength, walking speed, and sit-to-stand test were measured to mirror physical function. Radiographs of hands and feet, pain, and the modified 28 joint disease activity score (DAS28) were also assessed. RESULTS: Mean total HAQ was 1.08 (0.68) in women and 0.67 (0.70) in men (p = 0.0031). Women had greater disability than men in five of the eight subdimensions of the HAQ. Grip strength was 48%, knee extension strength 46%, trunk extension strength 54%, and trunk flexion strength 43% lower in women than in men. Knee extension strength was inversely correlated with walking time (r = -0.63 (95% confidence interval, -0.73 to -0.51)) and with sit-to-stand test (r = -0.47 (-0.60 to -0.31)). In an ordered logistic regression analysis in female rheumatoid patients, DAS28, pain, knee extension strength, and grip strength were associated with the total HAQ disability index. CONCLUSIONS: Women reported greater disability than men both in the total HAQ and in the majority of its eight subdimensions. In addition to disease activity and pain, muscle strength has a major impact on disability especially in female rheumatoid patients. PMID- 15901636 TI - MicroRNA Mirn122a reduces expression of the posttranscriptionally regulated germ cell transition protein 2 (Tnp2) messenger RNA (mRNA) by mRNA cleavage. AB - MicroRNAs play important roles in regulating development at both transcriptional and posttranscriptional levels. Here, we report 29 microRNAs from mouse testis that are differentially expressed as the prepubertal testis differentiates to the adult testis. Using computational analyses to identify potential microRNA target mRNAs, we identify several possible male germ cell target mRNAs. One highly conserved sequence in the 3'-untranslated region (UTR) of transition protein 2 (Tnp2) mRNA, a testis-specific and posttranscriptionally regulated mRNA in postmeiotic germ cells, is complementary to Mirn122a. Mirn122a is enriched in late-stage male germ cells and is predominantly on polysomes. Mirn122a, but not another noncomplementary microRNA, inhibits the activity of a luciferase reporter construct containing the 3'-UTR of Tnp2. Site-directed mutations of Mirn122a indicate that base pairing of the 5'-region of Mirn122a to its complementary site in the 3'-UTR of Tnp2 mRNA is essential for the downregulation of luciferase activity. Real-time reverse transcription-polymerase chain reaction and ribonuclease protection assays reveal that the Mirn122a-directed decrease of the Tnp2 reporter gene activity results from mRNA cleavage. We propose that specific microRNAs, such as Mirn122a, could be involved in the posttranscriptional regulation of mRNAs such as Tnp2 in the mammalian testis. PMID- 15901637 TI - Effects of embryo culture on angiogenesis and morphometry of bovine placentas during early gestation. AB - The objective of this study was to determine the effects of undefined and semidefined culture systems for in vitro embryo production on angiogenesis and morphometry of bovine placentas during early gestation. Blastocysts produced in vivo were recovered from superovulated Holstein cows and served as controls. Blastocysts produced in vitro were exposed to either serum-supplemented medium with cumulus cell coculture (in vitro-produced with serum; IVPS) or modified synthetic oviductal fluid medium without serum or coculture (mSOF). Single blastocysts from each production system were transferred into heifers. Fetuses and placentas were recovered on Day 70 of gestation. Cotyledonary tissues were obtained for quantification of vascular endothelial growth factor (VEGF) and peroxisome proliferator-activated receptor-gamma (PPARG) mRNA and protein. Samples of placentomes were prepared for immunocytochemistry and histological analysis. Placentas from the mSOF group were heavier and had the fewest placentomes, least placental fluid, and lowest placental efficiency (fetal weight/placental weight) compared with the in vivo and IVPS groups. There was no effect of embryo culture system on volume densities of fetal villi or maternal endometrium within placentomes. The volume density of fetal pyknotic cells was increased in placentomes in the mSOF group compared with the in vivo and IVPS groups. Placentomes in the mSOF group had decreased densities of blood vessels and decreased levels of VEGF mRNA in cotyledonary tissue. In conclusion, compared with placentas from embryos produced in vivo or in vitro using an undefined culture system, placentas from embryos produced in vitro using a semidefined culture system exhibited a greater degree of aberrant development of the placenta during early gestation. PMID- 15901638 TI - Effects of progranulin on blastocyst hatching and subsequent adhesion and outgrowth in the mouse. AB - Using cDNA microarray methodology, we have shown previously that transcripts of progranulin gene (Grn, also known as acrogranin), a recently identified autocrine growth factor, were upregulated in mouse blastocysts adhered to the filter membrane in an in vitro-culture system. In the present study, we investigated the expression and effects of progranulin on blastocyst hatching, adhesion, and embryo outgrowth during the peri-implantation period in the mouse. During this period, substantial amounts of Grn mRNA were present in both inner cell mass (ICM) and trophectoderm. Progranulin was localized exclusively to the surface of the trophectoderm in early and pre- and postadhesion blastocysts as well as in trophoblast cells and ICM of outgrowth embryos, being secreted as a single, 88 kDa form into the surrounding medium. NIH3T3 cells that had been transfected with a progranulin expression construct secreted the 88-kDa form of the protein, from which a 68-kDa form could be generated by deglycosylation. In vitro treatment of blastocysts with recombinant progranulin promoted blastocyst hatching, adhesion, and outgrowth, whereas rabbit anti-mouse progranulin immunoglobulin G reduced the incidence of blastocyst hatching, adhesion, and outgrowth. Studies of bromodeoxyuridine incorporation and immunodissection of the ICM revealed that progranulin was effective on the trophectoderm but not on the ICM. These results indicate that progranulin is an important factor for the processes of blastocyst hatching, adhesion, and outgrowth, and they suggest that the effects of progranulin on blastocyst adhesion and outgrowth may have been triggered by the previous action of progranulin to induce hatching of the blastocysts. PMID- 15901639 TI - Demonstration of a glycoprotein derived from the Ceacam10 gene in mouse seminal vesicle secretions. AB - CEACAM10 was purified from mouse seminal vesicle secretions by a series of purification steps that included ion exchange chromatography on a DEAE-Sephacel column and ion exchange high-performance liquid chromatography on a sulfopropyl column. It was shown to be a 36-kDa glycoprotein with an N-linked carbohydrate moiety. The circular dichromoism spectrum of CEACAM10 in 50 mM phosphate buffer at pH 7.4 appeared as one negative band arising from the beta form at 217 nm. CEACAM10 was expressed predominantly in seminal vesicles of adult mice. Both CEACAM10 and its mRNA were demonstrated on the luminal epithelium of the mucosal folds in the seminal vesicle. The amount of Ceacam10 mRNA in the seminal vesicle was correlated with the stage of animal maturation. Castration of adult mice resulted in cessation of Ceacam10 expression, while treatment of castrated mice with testosterone propionate in corn oil restored Ceacam10 expression in the seminal vesicle. During the entire course of pregnancy, Ceacam10 might be silent in the embryo. A cytochemical study illustrated the presence of the CEACAM10 binding region on the entire surface of mouse sperm. CEACAM10-sperm binding greatly enhanced sperm motility in vitro. PMID- 15901640 TI - Identification of a new variant of PDE1A calmodulin-stimulated cyclic nucleotide phosphodiesterase expressed in mouse sperm. AB - In mature sperm, cAMP plays an important role as a second messenger regulating functions that include capacitation, the acrosome reaction, motility, and, in some cases, chemosensing. We have cloned from mouse testis a novel calmodulin stimulated cyclic nucleotide phosphodiesterase 1A isoform, Pde1a_v7 (mmPDE1A7), which arises from an alternative transcription start in the cyclic nucleotide phosphodiesterase 1A gene. The open reading frame is predicted to encode a polypeptide with a molecular mass of 52 kDa. Two further variants of this form, which contain two additional new exons, arise from alternative splicing. Analysis of testis cDNA by real-time polymerase chain reaction (PCR) indicates that the Pde1A_v7 transcript variant is the most abundant. The PDE1A_v7 protein uniquely lacks the first amino-terminal calmodulin-binding domain, but does possess an inhibitory domain and a second calmodulin-binding site shared with other variants. In vitro translation of the corresponding Pde1a_v7 cDNA produced a 52 kDa polypeptide having cyclic nucleotide hydrolytic activity, which was stimulated threefold by calcium-bound calmodulin. Immunoprecipitation of cyclic nucleotide phosphodiesterase 1 activity from detergent extracts of mouse sperm revealed a major protein of the size expected for PDE1A_v7, and the immunocytochemical staining for cyclic nucleotide phosphodiesterase 1A in mouse sperm showed intense immunoreactivity in the tail only. These observations, along with the PCR data, strongly suggest that this new variant PDE1A_v7 is the major form of cyclic nucleotide phosphodiesterase 1A expressed in mature sperm and is therefore likely to play an important role in cyclic nucleotide regulation of mature sperm function. PMID- 15901641 TI - Stimulation of macrophage migration inhibitory factor expression in endometrial stromal cells by interleukin 1, beta involving the nuclear transcription factor NFkappaB. AB - Endometriosis, the ectopic development of endometrial tissue, is, particularly in peritoneal endometriosis, believed to result from tubal reflux of menstrual tissue. The release of cytokines and growth factors by refluxed endometrial cells in response to peritoneal inflammatory stimuli may enhance the capability of endometrial cells to implant and grow into the peritoneal host tissue. Herein we report that interleukin 1 (IL1), a major proinflammatory cytokine that is overproduced by endometriosis women-derived peritoneal macrophages and found in elevated concentrations in the peritoneal fluid of patients with endometriosis, stimulates the synthesis and the secretion of macrophage migration inhibitory factor (MIF) by human endometrial stromal cells. IL1B (0.1-100 ng/ml) exerted dose- and time-dependent effects of MIF protein secretion and mRNA synthesis, as shown by ELISA and reverse transcription-polymerase chain reaction, respectively. IL1B appeared to induce MIF gene transcription via the kappaB nuclear transcription factor (NFkappaB), as shown by electrophoretic mobility shift assay and Western blot analysis of IkappaB phosphorylation. Curcumin (10(-8) M), which is known for inhibiting NFkappaB activation, inhibited IL1B-induced MIF secretion as well as NFkappaB nuclear translocation and DNA binding. Taken together, these findings clearly show that IL1B up-regulates the expression of MIF in endometrial stromal cells in vitro and acts via NFkappaB. This may play an important role in the physiology of the human endometrium and the pathophysiology of endometriosis considering the immunomodulatory properties of MIF as well as its role in cell growth, angiogenesis and tissue remodeling. PMID- 15901642 TI - Exposure in utero to di(n-butyl) phthalate alters the vimentin cytoskeleton of fetal rat Sertoli cells and disrupts Sertoli cell-gonocyte contact. AB - Di(n-butyl) phthalate (DBP) is commonly used in personal care products and as a plasticizer to soften consumer plastic products. Male rats exposed to DBP in utero have malformations of the male reproductive tract and testicular atrophy characterized by degeneration of seminiferous epithelium and decreased sperm production. In the fetal testis, in utero exposure to DBP reportedly resulted in reduced testosterone levels, Leydig cell aggregates, and multinucleated gonocytes (MNG). We investigated whether exposure in utero to DBP affects rat fetal Sertoli cells and compromises interactions between Sertoli and germ cells in the developing testis. Histological examination showed that MNG occurred at low frequency in the normal fetal rat testis. Exposure in utero at the dose level of DBP above estimated environmental or occupational human exposure levels significantly increased the number of these abnormal germ cells. Postnatally, MNG exhibited aberrant mitoses and were detected at the basal lamina. MNG were not apoptotic in the fetal and postnatal rat testes, as indicated by TUNEL. Sertoli cells in DBP-exposed fetal testis had retracted apical processes, altered organization of the vimentin cytoskeleton, and abnormal cell-cell contacts with gonocytes. The effect of DBP on Sertoli cell morphology at the level of light microscopy was reversed after birth and cessation of exposure. Our data indicate that fetal Sertoli cells are targeted by exposure in utero to DBP and suggest that abnormal interactions between Sertoli and germ cells during fetal life play a role in the development of MNG. PMID- 15901643 TI - Systematic review and meta-analysis of studies of the timing of tracheostomy in adult patients undergoing artificial ventilation. AB - OBJECTIVE: To compare outcomes in critically ill patients undergoing artificial ventilation who received a tracheostomy early or late in their treatment. DATA SOURCES: The Cochrane Central Register of Clinical Trials, Medline, Embase, CINAHL, the National Research Register, the NHS Trusts Clinical Trials Register, the Medical Research Council UK database, the NHS Research and Development Health Technology Assessment Programme, the British Heart Foundation database, citation review of relevant primary and review articles, and expert informants. STUDY SELECTION: Randomised and quasi-randomised controlled studies that compared early tracheostomy with either late tracheostomy or prolonged endotracheal intubation. From 15,950 articles screened, 12 were identified as "randomised or quasi randomised" controlled trials, and five were included for data extraction. DATA EXTRACTION: Five studies with 406 participants were analysed. Descriptive and outcome data were extracted. The main outcome measure was mortality in hospital. The incidence of hospital acquired pneumonia, length of stay in a critical care unit, and duration of artificial ventilation were also recorded. Random effects meta-analyses were performed. RESULTS: Early tracheostomy did not significantly alter mortality (relative risk 0.79, 95% confidence interval 0.45 to 1.39). The risk of pneumonia was also unaltered by the timing of tracheostomy (0.90, 0.66 to 1.21). Early tracheostomy significantly reduced duration of artificial ventilation (weighted mean difference -8.5 days, 95% confidence interval -15.3 to -1.7) and length of stay in intensive care (-15.3 days, -24.6 to -6.1). CONCLUSIONS: In critically ill adult patients who require prolonged mechanical ventilation, performing a tracheostomy at an earlier stage than is currently practised may shorten the duration of artificial ventilation and length of stay in intensive care. PMID- 15901644 TI - Adult socioeconomic, educational, social, and psychological outcomes of childhood obesity: a national birth cohort study. AB - OBJECTIVES: To assess adult socioeconomic, educational, social, and psychological outcomes of childhood obesity by using nationally representative data. DESIGN: 1970 British birth cohort. PARTICIPANTS: 16,567 babies born in Great Britain 5-11 April 1970 and followed up at 5, 10, and 29-30 years. MAIN OUTCOME MEASURES: Obesity at age 10 and 30 years. Self reported socioeconomic, educational, psychological, and social outcomes at 30 years. Odds ratios were calculated for the risk of each adult outcome associated with obesity in childhood only, obesity in adulthood only, and persistent child and adult obesity, compared with those obese at neither period. RESULTS: Of the 8490 participants with data on body mass index at 10 and 30 years, 4.3% were obese at 10 years and 16.3% at 30 years. Obesity in childhood only was not associated with adult social class, income, years of schooling, educational attainment, relationships, or psychological morbidity in either sex after adjustment for confounding factors. Persistent obesity was not associated with any adverse adult outcomes in men, though it was associated among women with a higher risk of never having been gainfully employed (odds ratio 1.9, 95% confidence interval 1.1 to 3.3) and not having a current partner (2.0, 1.3 to 3.3). CONCLUSIONS: Obesity limited to childhood has little impact on adult outcomes. Persistent obesity in women is associated with poorer employment and relationship outcomes. Efforts to reduce the socioeconomic and psychosocial burden of obesity in adult life should focus on prevention of the persistence of obesity from childhood into adulthood. PMID- 15901645 TI - Motor neuron pathology in experimental autoimmune encephalomyelitis: studies in THY1-YFP transgenic mice. AB - Using adult male C57BL/6 mice that express a yellow fluorescent protein transgene in their motor neurons, we induced experimental autoimmune encephalomyelitis (EAE) by immunization with myelin oligodendrocyte glycoprotein peptide 35-55 (MOG peptide) in complete Freund's adjuvant (CFA). Control mice of the same transgenic strain received CFA without MOG peptide. Early in the course of their illness, the EAE mice showed lumbosacral spinal cord inflammation, demyelination and axonal fragmentation. By 14 weeks post-MOG peptide, these abnormalities were much less prominent, but the mice remained weak and, as in patients with progressive multiple sclerosis, spinal cord atrophy had developed. There was no significant loss of lumbar spinal cord motor neurons in the MOG peptide-EAE mice. However, early in the course of the illness, motor neuron dendrites were disrupted and motor neuron expression of hypophosphorylated neurofilament-H (hypoP-NF-H) immunoreactivity was diminished. By 14 weeks post-MOG peptide, hypoP-NF-H expression had returned to normal, but motor neuron dendritic abnormalities persisted and motor neuron perikaryal atrophy had appeared. We hypothesize that these motor neuron abnormalities contribute to weakness in this form of EAE and speculate that similar motor neuron abnormalities are present in patients with progressive multiple sclerosis. PMID- 15901646 TI - A longitudinal fMRI study on motor activity in patients with multiple sclerosis. AB - Using functional MRI (fMRI), patients with multiple sclerosis showed a greater extent of motor activation than controls. Although functional changes are often interpreted as adaptive and as a contributing factor in limiting the clinical deficit, no longitudinal studies have yet been performed for multiple sclerosis. Sixteen patients with multiple sclerosis, two patients with possible multiple sclerosis and nine age-matched controls underwent two fMRI studies with a time interval of 15-26 months. The motor task consisted of a self-paced sequential finger opposition movement with the right hand. Patients with multiple sclerosis exhibited greater bilateral activation than controls in both fMRI studies. At follow-up, patients showed a reduction in functional activity in the ipsilateral sensorimotor cortex and in the contralateral cerebellum. No significant differences between the two fMRI studies were observed in controls. Activation changes in ipsilateral motor areas correlated inversely with age, extent and progression of T1 lesion load, and occurrence of a new relapse. This study may help the understanding of the evolution of brain plastic changes in multiple sclerosis indicating that, in younger patients with a less structural brain damage and benign clinical course, the brain reorganizes its functional activity towards a more lateralized pattern of brain activation. The tendency towards a normalization of brain functional activity is hampered in older patients and in those developing relapses or new irreversible brain damage. PMID- 15901647 TI - Ion changes in spreading ischaemia induce rat middle cerebral artery constriction in the absence of NO. AB - In rats, cortical spreading hyperaemia is coupled to a spreading neuroglial depolarization wave (spreading depression) under physiological conditions, whereas cortical spreading ischaemia is coupled to it if red blood cell products are present in the subarachnoid space. Spreading ischaemia has been proposed as the pathophysiological correlate of the widespread cortical infarcts abundantly found in autopsy studies of patients with subarachnoid haemorrhage. The purpose of the present study was to investigate whether the extracellular ion changes associated with the depolarization wave may cause the vasoconstriction underlying spreading ischaemia. We induced spreading ischaemia in vivo with the nitric oxide (NO) scavenger oxyhaemoglobin and an elevated K+ concentration in the subarachnoid space while slow potential, pH, extracellular volume and concentrations of K+, Na+, Ca2+ and Cl- were measured in the cortex with microelectrodes. We then extraluminally applied an ionic cocktail (cocktail(SI)) to the isolated middle cerebral artery in vitro, matching the ionic composition of the extracellular space as measured during spreading ischaemia in vivo. Extraluminal application of cocktail(SI) caused middle cerebral artery dilatation in the absence and constriction in the presence of NO synthase inhibition in vitro, corresponding with the occurrence of spreading hyperaemia in the presence and spreading ischaemia in the absence of NO in vivo. The L-type Ca2+ inhibitor nimodipine caused the cocktail(SI)-induced vasoconstriction to revert to vasodilatation in the absence of NO in vitro similar to the reversal of spreading ischaemia to spreading hyperaemia in response to nimodipine in vivo. We found that K+ was the predominant vasoconstrictor contained in cocktail(SI). Its vasoconstrictor action was augmented by NO synthase inhibition. Our results suggest that, under elevated baseline K+ as a hallmark of any condition of energy deficiency, the extracellular ion changes represent the essential mediator of the vascular response to spreading neuroglial depolarization. In the presence of NO they mediate vasodilatation and in its absence they mediate constriction. PMID- 15901648 TI - Brain biopsy in dementia. AB - Brain biopsy has an uncertain role in the diagnosis of dementia. Here we report a retrospective analysis of 90 consecutive cerebral biopsies undertaken for the investigation of dementia in adults at a tertiary referral centre between 1989 and 2003. In most cases (90%), biopsy consisted of a right frontal full thickness resection of cortex, white matter and overlying leptomeninges. Fifty-seven per cent of biopsies were diagnostic: the most frequent diagnoses were Alzheimer's disease (18%), Creutzfeldt-Jakob disease (12%) and inflammatory disorders (9%). Other diagnoses in individual patients included Pick's disease, corticobasal degeneration and other tauopathies, Lewy body dementia, multiple sclerosis, Whipple's disease, progressive multifocal leucoencephalopathy, cerebral autosomal dominant arteriopathy with subcortical ischaemic leucoencephalopathy, vasculopathies and paraneoplastic encephalopathy. The most frequent biopsy finding in the non-diagnostic group and for the series as a whole (37%) was non specific gliosis variably affecting both cortex and white matter. Complications (11%) included seizures, intracranial and wound infections, and intracranial haemorrhage; there were no deaths or lasting neurological sequelae attributable to the procedure. No trends in diagnostic yield or complication rate over the course of the series were identified. Information obtained at biopsy determined treatment in 11%. A raised cerebrospinal fluid cell count was the only robust predictor of a potentially treatable (inflammatory) process at biopsy. The constellation of behavioural change, raised CSF protein and matched oligoclonal bands in CSF and serum was associated with non-specific gliosis at biopsy. This series underlines the value of cerebral biopsy in the diagnosis of dementia, and suggests that certain clinical and laboratory features may be useful in guiding the decision to proceed to brain biopsy where a treatable disease cannot be excluded by other means. PMID- 15901649 TI - Mitochondrial changes in skeletal muscle in amyotrophic lateral sclerosis and other neurogenic atrophies. AB - Previous findings suggested specific mitochondrial dysfunction in skeletal muscle of patients with amyotrophic lateral sclerosis (ALS). To answer the question of whether the dysfunction is specific, we investigated the histochemical distribution of mitochondrial marker activities, the ratio of mitochondrial (mt) versus nuclear (n) DNA, and the activities of citrate synthase (CS) and respiratory chain enzymes in muscle biopsies of 24 patients with sporadic ALS. The data were compared with those in 23 patients with other neurogenic atrophies (NAs), and 21 healthy controls. Muscle histology revealed similar signs of focally diminished mitochondrial oxidation activity in muscle fibres in both diseased groups. There was only minimal decline of mt/nDNA ratios in ALS and NA patients in comparison with healthy controls. The specific activities of mitochondrial markers CS and succinate dehydrogenase were significantly increased in both ALS and NA patients. The specific activities of respiratory chain enzymes were not significantly different in all three groups. It is concluded that the histochemical, biochemical and molecular mitochondrial changes in muscle are not specific for ALS, but accompany other NAs as well. PMID- 15901650 TI - Temporal analysis of cortical mechanisms for pain relief by tactile stimuli in humans. AB - The mechanisms by which vibrotactile stimuli relieve pain are not well understood, especially in humans. We recorded cortical magnetic responses to paired noxious (intra-epidermal electrical stimulation, IES) and innocuous (transcutaneous electrical stimulation, TS) stimuli applied to the back at a conditioning-test interval (CTI) of -500 to 500 ms. Results showed that IES induced responses were remarkably attenuated when TS was applied 20-60 ms later and 0-500 ms earlier than IES (CTI = -60 to 500 ms). Since the signals evoked by IES reached the spinal cord (CTI = -60 to -20 ms conditions) and the cortex (-60 and -40 ms condition) earlier than those evoked by TS, the present results indicate that cortical responses to noxious stimuli can be inhibited by innocuous tactile stimuli at the cortical level, with minimal contribution at the spinal level. PMID- 15901651 TI - Parasagittal asymmetries of the corpus callosum. AB - Significant relationships have been reported between midsagittal areas of the corpus callosum and the degree of interhemispheric transfer, functional lateralization and structural brain asymmetries. No study, however, has examined whether parasagittal callosal asymmetries (i.e. those close to the midline of the brain), which may be of specific functional consequence, are present in the human brain. Thus, we applied magnetic resonance imaging and novel computational surface-based methods to encode hemispheric differences in callosal thickness at a very high resolution. Discrete callosal areas were also compared between the hemispheres. Furthermore, acknowledging the frequently reported sex differences in callosal morphology, parasagittal callosal asymmetries were examined within each gender. Results showed significant rightward asymmetries of callosal thickness predominantly in the anterior body and anterior third of the callosum, suggesting a more diffuse functional organization of callosal projections in the right hemisphere. Asymmetries were increased in men, supporting the assumption of a sexually dimorphic organization of male and female brains that involves hemispheric relations and is reflected in the organization and distribution of callosal fibers. PMID- 15901652 TI - Reflexive orienting in spatial neglect is biased towards behaviourally salient stimuli. AB - Patients with spatial neglect are impaired when detecting contralesional targets presented shortly after an ipsilesional cue. This "disengagement" deficit is believed to reflect reflexive orienting towards ipsilesional stimuli that is independent of behavioural goals. Here, we show that the extent of this spatial bias depends on the behavioural salience of ipsilesional stimuli. Healthy participants, brain-injured patients without neglect and neglect patients reacted to ipsilesional and contralesional visual targets. Prior to target presentation, a visual cue similar or dissimilar to the target was presented at target position or opposite the target. Although participants did not react to the similar cue, it had high behavioural salience since it shared features with the target stimulus. Neglect patients showed dramatically increased reaction times to contralesional targets, but only when these followed behaviourally relevant ipsilesional cues. No decrease of performance was observed with irrelevant cues. This performance pattern was not due to perceptual similarity, since the same effect was found when cue and target were semantically related but differed perceptually. Importantly, semantically related cues ceased to attract attention when they were defined as behaviourally irrelevant. These results show that neglect patients only orient attention reflexively towards ipsilesional stimuli with high behavioural salience. PMID- 15901653 TI - Running wheel accessibility affects the regional electroencephalogram during sleep in mice. AB - Regional aspects of sleep homeostasis were investigated in mice provided with a running wheel for several weeks. Electroencephalogram (EEG) spectra of the primary motor (frontal) and somatosensory cortex (parietal) were recorded for three consecutive days. On a single day (day 2) the wheel was locked to prevent running. Wheel running correlated negatively with the frontal-parietal ratio of slow-wave activity (EEG power between 0.75 and 4.0 Hz) in the first 2 h after sleep onset (r = -0.60; P < 0.01). On day 2 frontal EEG power (2.25-8.0 Hz) in non-rapid eye movement sleep exceeded the level of the previous day, indicating that the diverse behaviors replacing wheel-running elicited more pronounced regional EEG differences. The frontal-parietal power ratio of the lower frequency bin (0.75-1.0 Hz) in the first 2 h of sleep after dark onset correlated positively with the duration of the preceding waking (r = 0.64; P < 0.001), whereas the power ratio in the remaining frequencies of the delta band (1.25-4.0 Hz) was unrelated to waking. The data suggest that in mice EEG power in the lower frequency, corresponding to the slow oscillations described in cats and humans, is related to local sleep homeostasis. PMID- 15901654 TI - Postnatal expression profile of OBCAM implies its involvement in visual cortex development and plasticity. AB - This study examined the expression of a neuron-specific cell adhesion molecule, OBCAM (opioid-binding cell adhesion molecule), at both the mRNA and protein levels in the cat primary visual cortex at various postnatal ages, using cDNA array analysis and immunocytochemistry. Results obtained using both methods showed that the expression level of OBCAM was high in young and low in older and adult visual cortex. OBCAM-immunoreactivities were associated predominantly with perikarya and dendrites of pyramidal neurons, and OBCAM-immunopositive neurons were present in all cortical layers. Immunostaining of OBCAM in adult visual cortex showed a reduced number of immunopositive neurons and neurites and relatively lower staining intensities as compared with younger animals. In addition, the number of OBCAM-immunopositive neurons was significantly higher in the visual cortex of 4-month-old animals dark-reared from birth than those in age matched normally reared animals. These results suggest that OBCAM may play an important role in visual cortex development and plasticity. PMID- 15901655 TI - Attentional modulation of cortical neuromagnetic gamma response to biological movement. AB - Processing of biological motion represented solely by a set of lights on the joints of a human body is traditionally viewed as largely independent of attention. Here, by manipulating attention-related task demands, we assess changes in the neuromagnetic cortical response to a point-light walker. Irrespective of task demands, biological motion evokes an increase in oscillatory gamma activity over the left parieto-occipital region at 80 ms post-stimulus. Only an attended walker, however, yielded further peaks over the right parietal (120 ms) and temporal (155 ms) cortices. By contrast, the magnetoencephalographic (MEG) response to an ignored walker is restricted to the left parieto-occipital region. In addition, peaks in oscillatory activity occur in response to both attended (canonical and scrambled) configurations at 180-200 ms from stimulus onset over the right fronto-temporal regions, most likely reflecting maintenance of the target configuration in working memory. For the first time, we demonstrate that the time course and topographic dynamics of oscillatory gamma activity in response to biological movement undergoes top-down influences and can be profoundly modulated by the withdrawal of attention. PMID- 15901656 TI - Repeated stress induces dendritic spine loss in the rat medial prefrontal cortex. AB - The prefrontal cortex (PFC) plays an important role in higher cognitive processes, and in the regulation of stress-induced hypothalamic-pituitary-adrenal (HPA) activity. Here we examined the effect of repeated restraint stress on dendritic spine number in the medial PFC. Rats were perfused after receiving 21 days of daily restraint stress, and intracellular iontophoretic injections of Lucifer Yellow were carried out in layer II/III pyramidal neurons in the anterior cingulate and prelimbic cortices. We found that stress results in a significant (16%) decrease in apical dendritic spine density in medial PFC pyramidal neurons, and confirmed a previous observation that total apical dendritic length is reduced by 20% in the same neurons. We estimate that nearly one-third of all axospinous synapses on apical dendrites of pyramidal neurons in medial PFC are lost following repeated stress. A decrease in medial PFC dendritic spines may not only be indicative of a decrease in the total population of axospinous synapses, but may impair these neurons' capacity for biochemical compartmentalization and plasticity in which dendritic spines play a major role. Dendritic atrophy and spine loss may be important cellular features of stress-related psychiatric disorders where the PFC is functionally impaired. PMID- 15901657 TI - Inhibitory and excitatory effects of iodobenzene on the antennal benzoic acid receptor cells of the female silk moth Bombyx mori L. AB - As shown in single-sensillum recordings, iodobenzene has a bimodal effect on the receptor cell tuned to benzoic acid (BA) of the female silk moth Bombyx mori. Exposure to iodobenzene causes an inhibition of the response to BA. With stimulation by iodobenzene alone, a reduction of basic nerve impulse firing during exposure is followed by a transient post-stimulus excitation (rebound). We suggest that inhibition suppresses excitation during exposure but fades afterwards more rapidly than excitation. Due to the spatial equivalence of the iodine and the acid residue, these effects might indicate opposing interactions of iodobenzene with the specific site for the key compound BA. This is supported by the fact that substitutions by smaller halogens are less effective in both inhibition and rebound. The inhibitory effect but not the rebound with iodobenzene alone was also observed in receptor cells tuned to key compounds other than benzoic acid, e.g. in the cell most sensitive to 2,6-dimethyl-5 heptene-2-ol (DMH-cell) occurring in the same sensillum as the BA-cell, or in the bombykol- and bombykal-cells of the male. At least in these cells the inhibitory effect might reflect the action of iodobenzene on a general site, e.g. the lipid matrix of the plasma membrane of the receptor cells. PMID- 15901658 TI - Written language of deaf and hard-of-hearing students in public schools. AB - We obtained data on the writing of 110 deaf or hard-of-hearing students attending public schools who completed the spontaneous writing portion of the Test of Written Language. The average written quotient for the sample was in the below average range but within 1 standard deviation of the test mean. Forty-nine percent of the sample received written quotients within or above the average range. Mean scores for the three subtests of contextual conventions, contextual language, and story construction were within the low-average range; between 55% and 68% of students scored within the average or above-average range for the subtests. Predictors of writing quotients were eligibility for free lunch, grade, degree of hearing loss and gender; however, only 18% of the variance in total writing quotients was explained by these variables. The data indicate that attention needs to be paid to the writing ability and instruction of many public school students regardless of degree of hearing loss. PMID- 15901659 TI - Retinoic acid regulates postnatal neurogenesis in the murine subventricular zone olfactory bulb pathway. AB - Neurogenesis persists throughout life in the rodent subventricular zone (SVZ) olfactory bulb pathway. The molecular regulation of this neurogenic circuit is poorly understood. Because the components for retinoid signaling are present in this pathway, we examined the influence of retinoic acid (RA) on postnatal SVZ olfactory bulb neurogenesis. Using both SVZ neurosphere stem cell and parasagittal brain slice cultures derived from postnatal mouse, we found that RA exposure increased neurogenesis by enhancing the proliferation and neuronal differentiation of forebrain SVZ neuroblasts. The RA precursor retinol had a similar effect, which was reversed by treating cultures with the RA synthesis inhibitor disulfiram. Electroporation of dominant-negative retinoid receptors into the SVZ of slice cultures also blocked neuroblast migration to the olfactory bulb and altered the morphology of the progenitors. Moreover, the administration of disulfiram to neonatal mice decreased in vivo cell proliferation in the striatal SVZ. These results indicate that RA is a potent mitogen for SVZ neuroblasts and is required for their migration to the olfactory bulb. The regulation of multiple steps in the SVZ-olfactory bulb neurogenic pathway by RA suggests that manipulation of retinoid signaling is a potential therapeutic strategy to augment neurogenesis after brain injury. PMID- 15901660 TI - Xema, a foxi-class gene expressed in the gastrula stage Xenopus ectoderm, is required for the suppression of mesendoderm. AB - The molecular basis of vertebrate germ layer formation has been the focus of intense scrutiny for decades, and the inductive interactions underlying this process are well defined. Only recently, however, have studies demonstrated that the regulated inhibition of ectopic germ layer formation is also crucial for patterning the early vertebrate embryo. We report here the characterization of Xema (Xenopus Ectodermally-expressed Mesendoderm Antagonist), a novel member of the Foxi-subclass of winged-helix transcription factors that is involved in the suppression of ectopic germ layer formation in the frog, Xenopus laevis. Xema transcripts are restricted to the animal pole ectoderm during early Xenopus development. Ectopic expression of Xema RNA inhibits mesoderm induction, both by growth factors and in the marginal zone, in vivo. Conversely, introduction of antisense morpholino oligonucleotides directed against the Xema transcript stimulates the expression of a broad range of mesodermal and endodermal marker genes in the animal pole. Our studies demonstrate that Xema is both necessary and sufficient for the inhibition of ectopic mesendoderm in the cells of the presumptive ectoderm, and support a model in which Fox proteins function in part to restrict inappropriate germ layer development throughout the vertebrate embryo. PMID- 15901661 TI - CrebA regulates secretory activity in the Drosophila salivary gland and epidermis. AB - Understanding how organs acquire the capacity to perform their respective functions is important for both cell and developmental biology. Here, we have examined the role of early-expressed transcription factors in activating genes crucial for secretory function in the Drosophila salivary gland. We show that expression of genes encoding proteins required for ER targeting and translocation, and proteins that mediate transport between the ER and Golgi is very high in the early salivary gland. This high level expression requires two early salivary gland transcription factors; CrebA is required throughout embryogenesis and Fkh is required only during late embryonic stages. As Fkh is required to maintain late CrebA expression in the salivary gland, Fkh probably works through CrebA to affect secretory pathway gene expression. In support of these regulatory interactions, we show that CrebA is important for elevated secretion in the salivary gland. Additionally, CrebA is required for the expression of the secretory pathway genes in the embryonic epidermis, where CrebA had previously been shown to be essential for cuticle development. We show that zygotic mutations in several individual secretory pathway genes result in larval cuticle phenotypes nearly identical to those of CrebA mutants. Thus, CrebA activity is linked to secretory function in multiple tissues. PMID- 15901662 TI - Sequential roles for Mash1 and Ngn2 in the generation of dorsal spinal cord interneurons. AB - The dorsal spinal cord contains a diverse array of neurons that connect sensory input from the periphery to spinal cord motoneurons and brain. During development, six dorsal neuronal populations (dI1-dI6) have been defined by expression of homeodomain factors and position in the dorsoventral axis. The bHLH transcription factors Mash1 and Ngn2 have distinct roles in specification of these neurons. Mash1 is necessary and sufficient for generation of most dI3 and all dI5 neurons. Unexpectedly, dI4 neurons are derived from cells expressing low levels or no Mash1, and this population increases in the Mash1 mutant. Ngn2 is not required for any specific neuronal cell type but appears to modulate the composition of neurons that form. In the absence of Ngn2, there is an increase in the number of dI3 and dI5 neurons, in contrast to the effects produced by activity of Mash1. Mash1 is epistatic to Ngn2, and, unlike the relationship between other neural bHLH factors, cross-repression of expression is not detected. Thus, bHLH factors, particularly Mash1 and related family members Math1 and Ngn1, provide a code for generating neuronal diversity in the dorsal spinal cord with Ngn2 serving to modulate the number of neurons in each population formed. PMID- 15901663 TI - Wnt2b inhibits differentiation of retinal progenitor cells in the absence of Notch activity by downregulating the expression of proneural genes. AB - During the development of the central nervous system, cell proliferation and differentiation are precisely regulated. In the vertebrate eye, progenitor cells located in the marginal-most region of the neural retina continue to proliferate for a much longer period compared to the ones in the central retina, thus showing stem-cell-like properties. Wnt2b is expressed in the anterior rim of the optic vesicles, and has been shown to control differentiation of the progenitor cells in the marginal retina. In this paper, we show that stable overexpression of Wnt2b in retinal explants inhibited cellular differentiation and induced continuous growth of the tissue. Notably, Wnt2b maintained the undifferentiated progenitor cells in the explants even under the conditions where Notch signaling was blocked. Wnt2b downregulated the expression of multiple proneural bHLH genes as well as Notch. In addition, expression of Cath5 under the control of an exogenous promoter suppressed the negative effect of Wnt2b on neuronal differentiation. Importantly, Wnt2b inhibited neuronal differentiation independently of cell cycle progression. We propose that Wnt2b maintains the naive state of marginal progenitor cells by attenuating the expression of both proneural and neurogenic genes, thus preventing those cells from launching out into the differentiation cascade regulated by proneural genes and Notch. PMID- 15901664 TI - Tbx20 is essential for cardiac chamber differentiation and repression of Tbx2. AB - Tbx20, a member of the T-box family of transcriptional regulators, shows evolutionary conserved expression in the developing heart. In the mouse, Tbx20 is expressed in the cardiac crescent, then in the endocardium and myocardium of the linear and looped heart tube before it is restricted to the atrioventricular canal and outflow tract in the multi-chambered heart. Here, we show that Tbx20 is required for progression from the linear heart tube to a multi-chambered heart. Mice carrying a targeted mutation of Tbx20 show early embryonic lethality due to hemodynamic failure. A linear heart tube with normal anteroposterior patterning is established in the mutant. The tube does not elongate, indicating a defect in recruitment of mesenchyme from the secondary heart field, even though markers of the secondary heart field are not affected. Furthermore, dorsoventral patterning of the tube, formation of working myocardium, looping, and further differentiation and morphogenesis fail. Instead, Tbx2, Bmp2 and vinexin alpha (Sh3d4), genes normally restricted to regions of primary myocardium and lining endocardium, are ectopically expressed in the linear heart tube of Tbx20 mutant embryos. Because Tbx2 is both necessary and sufficient to repress chamber differentiation (Christoffels et al., 2004a; Harrelson et al., 2004), Tbx20 may ensure progression to a multi-chambered heart by repressing Tbx2 in the myocardial precursor cells of the linear heart tube destined to form the chambers. PMID- 15901665 TI - Identification of functional sine oculis motifs in the autoregulatory element of its own gene, in the eyeless enhancer and in the signalling gene hedgehog. AB - In Drosophila, the sine oculis (so) gene is important for the development of the entire visual system, including Bolwig's organ, compound eyes and ocelli. Together with twin of eyeless, eyeless, eyes absent and dachshund, so belongs to a network of genes that by complex interactions initiate eye development. Although much is known about the genetic interactions of the genes belonging to this retinal determination network, only a few such regulatory interactions have been analysed down to the level of DNA-protein interactions. Previous work in our laboratory identified an eye/ocellus specific enhancer of the sine oculis gene that is directly regulated by eyeless and twin of eyeless. We further characterized this regulatory element and identified a minimal enhancer fragment of so that sets up an autoregulatory feedback loop crucial for proper ocelli development. By systematic analysis of the DNA-binding specificity of so we identified the most important nucleotides for this interaction. Using the emerging consensus sequence for SO-DNA binding we performed a genome-wide search and have thereby been able to identify eyeless as well as the signalling gene hedgehog as putative targets of so. Our results strengthen the general assumption that feedback loops among the genes of the retinal determination network are crucial for proper development of eyes and ocelli. PMID- 15901666 TI - The role of apoptosis in the regulation of trophoblast survival and differentiation during pregnancy. AB - Apoptosis is important for normal placental development, but it may also be involved in the pathophysiology of pregnancy-related diseases. Normal placental development is dependent upon the differentiation and invasion of the trophoblast, the main cellular component of the placenta. Trophoblast apoptosis increases in normal placentas as gestation proceeds, and a greater incidence of trophoblast apoptosis has been observed in pregnancies complicated by preeclampsia or intrauterine growth retardation (IUGR). In response to different stimuli, apoptosis may be initiated extrinsically by the death receptor pathway or intrinsically by the mitochondrial pathway. The central executioners of apoptosis are the caspases, which cleave numerous vital cellular proteins to affect the apoptotic cascade. By inhibiting caspase activation, several endogenous inhibitors, including flice-like inhibitory proteins (FLIPs), inhibitors of apoptosis (IAPs), and antiapoptotic Bcl-2 family members, can prevent further propagation of the death signal. Macrophages present at the maternal-fetal interface may also contribute to trophoblast survival by removing apoptotic cells and producing cytokines and growth factors, which influence the progression of the apoptotic cascade. This review focuses on the role of apoptosis in trophoblast development and differentiation, the molecular mechanisms by which normal trophoblast apoptosis can occur, and how it is regulated to prevent excessive trophoblast apoptosis and possible pregnancy complications. PMID- 15901667 TI - The decline of androgen levels in elderly men and its clinical and therapeutic implications. AB - Aging in men is accompanied by a progressive, but individually variable decline of serum testosterone production, more than 20% of healthy men over 60 yr of age presenting with serum levels below the range for young men. Albeit the clinical picture of aging in men is reminiscent of that of hypogonadism in young men and decreased testosterone production appears to play a role in part of these clinical changes in at least some elderly men, the clinical relevancy of the age related decline in sex steroid levels in men has not been unequivocally established. In fact, minimal androgen requirements for elderly men remain poorly defined and are likely to vary between individuals. Consequently, borderline androgen deficiency cannot be reliably diagnosed in the elderly, and strict differentiation between "substitutive" and "pharmacological" androgen administration is not possible. To date, only a few hundred elderly men have received androgen therapy in the setting of a randomized, controlled study, and many of these men were not androgen deficient. Most consistent effects of treatment have been on body composition, but to date there is no evidence-based documentation of clinical benefits of androgen administration to elderly men with normal or moderately low serum testosterone in terms of diminished morbidity or of improved survival or quality of life. Until the long-term risk-benefit ratio for androgen administration to elderly is established in adequately powered trials of longer duration, androgen administration to elderly men should be reserved for the minority of elderly men who have both clear clinical symptoms of hypogonadism and frankly low serum testosterone levels. PMID- 15901668 TI - Transforming growth factor-beta1 to the bone. AB - TGF-beta1 is a ubiquitous growth factor that is implicated in the control of proliferation, migration, differentiation, and survival of many different cell types. It influences such diverse processes as embryogenesis, angiogenesis, inflammation, and wound healing. In skeletal tissue, TGF-beta1 plays a major role in development and maintenance, affecting both cartilage and bone metabolism, the latter being the subject of this review. Because it affects both cells of the osteoblast and osteoclast lineage, TGF-beta1 is one of the most important factors in the bone environment, helping to retain the balance between the dynamic processes of bone resorption and bone formation. Many seemingly contradictory reports have been published on the exact functioning of TGF-beta1 in the bone milieu. This review provides an overall picture of the bone-specific actions of TGF-beta1 and reconciles experimental discrepancies that have been reported for this multifunctional cytokine. PMID- 15901669 TI - Guidelines for the diagnosis and treatment of chronic heart failure: executive summary (update 2005): The Task Force for the Diagnosis and Treatment of Chronic Heart Failure of the European Society of Cardiology. PMID- 15901670 TI - PAX6 expression identifies progenitor cells for corneal keratocytes. AB - Keratocytes of the corneal stroma produce a transparent extracellular matrix required for vision. During wound-healing and in vitro, keratocytes proliferate, becoming fibroblastic, and lose biosynthesis of unique corneal matrix components. This study sought identification of cells in the corneal stroma capable of assuming a keratocyte phenotype after extensive proliferation. About 3% of freshly isolated bovine stromal cells exhibited clonal growth. In low-mitogen media, selected clonal cultures displayed dendritic morphology and expressed high levels of keratan sulfate, aldehyde dehydrogenase 3A1, and keratocan, molecular markers of keratocyte phenotype. In protein-free media, both primary keratocytes and selected clonal cells aggregated to form attachment-independent spheroids expressing elevated levels of those marker molecules. The selected clonal cells exhibited normal karyotype and underwent replicative senescence after 65-70 population doublings; however, they continued expression of keratocyte phenotypic markers throughout their replicative life span. The progenitor cells expressed elevated mRNA for several genes characteristic of stem cells and also for genes expressed during ocular development PAX6, Six2, and Six3. PAX6 protein was detected in the cultured progenitor cells and a small number of stromal cells in intact tissue but was absent in cultured keratocytes and fibroblasts. Cytometry demonstrated PAX6 protein in 4% of freshly isolated stromal cells. These results demonstrate the presence of a previously unrecognized population of PAX6-positive cells in adult corneal stroma that maintain the potential to assume a keratocyte phenotype even after extensive replication. The presence of such progenitor cells has implications for corneal biology and for cell-based therapies targeting corneal scarring. PMID- 15901671 TI - Association of valproate-induced teratogenesis with histone deacetylase inhibition in vivo. AB - Chemically induced birth defects are an important public health and human problem. Here we use Xenopus and zebrafish as models to investigate the mechanism of action of a well-known teratogen, valproic acid (VPA). VPA is a drug used in treatment of epilepsy and bipolar disorder but causes spina bifida if taken during pregnancy. VPA has several biochemical activities, including inhibition of histone deacetylases (HDACs). To investigate the mechanism of action of VPA, we compared its effects in Xenopus and zebrafish embryos with those of known HDAC inhibitors and noninhibitory VPA analogs. We found that VPA and other HDAC inhibitors cause very similar and characteristic developmental defects whereas VPA analogs with poor inhibitory activity in vivo have little teratogenic effect. Unbiased microarray analysis revealed that the effects of VPA and trichostatin A (TSA), a structurally unrelated HDAC inhibitor, are strikingly concordant. The concordance is apparent both by en masse correlation of fold-changes and by detailed similarity of dose-response profiles of individual genes. Together, the results demonstrate that the teratogenic effects of VPA are very likely mediated specifically by inhibition of HDACs. PMID- 15901672 TI - Proapoptotic Bak is sequestered by Mcl-1 and Bcl-xL, but not Bcl-2, until displaced by BH3-only proteins. AB - Commitment of cells to apoptosis is governed largely by the interaction between members of the Bcl-2 protein family. Its three subfamilies have distinct roles: The BH3-only proteins trigger apoptosis by binding via their BH3 domain to prosurvival relatives, while the proapoptotic Bax and Bak have an essential downstream role involving permeabilization of organellar membranes and induction of caspase activation. We have investigated the regulation of Bak and find that, in healthy cells, Bak associates with Mcl-1 and Bcl-x(L) but surprisingly not Bcl 2, Bcl-w, or A1. These interactions require the Bak BH3 domain, which is also necessary for Bak dimerization and killing activity. When cytotoxic signals activate BH3-only proteins that can engage both Mcl-1 and Bcl-x(L) (such as Noxa plus Bad), Bak is displaced and induces cell death. Accordingly, the BH3-only protein Noxa could bind to Mcl-1, displace Bak, and promote Mcl-1 degradation, but Bak-mediated cell death also required neutralization of Bcl-x(L) by other BH3 only proteins. The results indicate that Bak is held in check solely by Mcl-1 and Bcl-x(L) and induces apoptosis only if freed from both. The finding that different prosurvival proteins have selective roles has notable implications for the design of anti-cancer drugs that target the Bcl-2 family. PMID- 15901673 TI - Histone deposition protein Asf1 maintains DNA replisome integrity and interacts with replication factor C. AB - Chromatin assembly and DNA replication are temporally coupled, and DNA replication in the absence of histone synthesis causes inviability. Here we demonstrate that chromatin assembly factor Asf1 also affects DNA replication. In budding yeast cells lacking Asf1, the amounts of several DNA replication proteins, including replication factor C (RFC), proliferating cell nuclear antigen (PCNA), and DNA polymerase epsilon (Pol epsilon), are reduced at stalled replication forks. In contrast, DNA polymerase alpha (Pol alpha) accumulates to higher than normal levels at stalled forks in asf1Delta cells. Using purified, recombinant proteins, we demonstrate that RFC directly binds Asf1 and can recruit Asf1 to DNA molecules in vitro. We conclude that histone chaperone protein Asf1 maintains a subset of replication elongation factors at stalled replication forks and directly interacts with the replication machinery. PMID- 15901674 TI - The C. elegans homolog of the mammalian tumor suppressor Dep-1/Scc1 inhibits EGFR signaling to regulate binary cell fate decisions. AB - Protein phosphorylation by kinases and the subsequent dephosphorylation by phosphatases are key mechanisms that regulate intracellular signal transduction during development. Here, we report the identification of the receptor protein tyrosine phosphatase DEP-1 as a negative regulator of the Caenorhabditis elegans EGF receptor. DEP-1 amplifies in the developing vulva and the excretory system the small differences in the amount of EGF signal received by equivalent precursor cells to achieve binary cell fate decisions. During vulval development, DEP-1 inhibits EGFR signaling in the secondary cell lineage in parallel with the NOTCH-mediated lateral inhibition, while EGFR signaling simultaneously down regulates DEP-1 and NOTCH expression in the primary cell lineage. This regulatory network of inhibitors results in the full activation of the EGFR/RAS/MAPK pathway in the primary vulval cells and at the same time keeps the EGFR/RAS/MAPK pathway inactive in the adjacent secondary cells. Mammalian Dep-1/Scc1 functions as a tumor-suppressor gene in the intestinal epithelium. Thus, mutations in human Dep 1 may promote tumor formation through a hyperactivation of the EGF receptor. PMID- 15901675 TI - O-linked glycosylation in maize-expressed human IgA1. AB - O-Linked glycans vary between eukaryotic cell types and play an important role in determining a glycoprotein's properties, including stability, target recognition, and potentially immunogenicity. We describe O-linked glycan structures of a recombinant human IgA1 (hIgA1) expressed in transgenic maize. Up to six proline/hydroxyproline conversions and variable amounts of arabinosylation (Pro/Hyp + Ara) were found in the hinge region of maize-expressed hIgA1 heavy chain (HC) by using a combination of matrix-assisted laser-desorption ionization mass spectrometry (MALDI MS), chromatography, and amino acid analysis. Approximately 90% of hIgA1 was modified in this way. An average molar ratio of six Ara units per molecule of hIgA1 was revealed. Substantial sequence similarity was identified between the HC hinge region of hIgA1 and regions of maize extensin family of hydroxyproline-rich glycoproteins (HRGP). We propose that because of this sequence similarity, the HC hinge region of maize-expressed hIgA1 can become a substrate for posttranslational conversion of Pro to Hyp by maize prolyl hydroxylase(s) with the subsequent arabinosylation of the Hyp residues by Hyp glycosyltransferase(s) in the Golgi apparatus in maize endosperm tissue. The observation of up to six Pro/Hyp hydroxylations combined with extensive arabinosylation in the hIgA1 HC hinge region is well in agreement with the Pro/Hyp hydroxylation model and the Hyp contiguity hypothesis suggested earlier in literature for plant HRGP. For the first time, the extensin-like Hyp/Pro conversion and O-linked arabinosylation are described for a recombinant therapeutic protein expressed in transgenic plants. Our findings are of significance to the field of plant biotechnology and biopharmaceutical industry developing transgenic plants as a platform for the production of recombinant therapeutic proteins. PMID- 15901676 TI - Imino sugar inhibitors for treating the lysosomal glycosphingolipidoses. AB - The inherited metabolic disorders of glycosphingolipid (GSL) metabolism are a relatively rare group of diseases that have diverse and often neurodegenerative phenotypes. Typically, a deficiency in catabolic enzyme activity leads to lysosomal storage of GSL substrates and in many diseases, several other glycoconjugates. A novel generic approach to treating these diseases has been termed substrate reduction therapy (SRT), and the discovery and development of N alkylated imino sugars as effective and approved drugs is discussed. An understanding of the molecular mechanism for the inhibition of the key enzyme in GSL biosynthesis, ceramide glucosyltransferase (CGT) by N-alkylated imino sugars, has also lead to compound design for improvements to inhibitory potency, bioavailability, enzyme selectivity, and biological safety. Following a successful clinical evaluation of one compound, N-butyl-deoxynojirimycin [(NB DNJ), miglustat, Zavesca], for treating type I Gaucher disease, issues regarding the significance of side effects and CNS access have been addressed as exposure of drug to patients has increased. An alternative experimental approach to treat specific glycosphingolipid (GSL) lysosomal storage diseases is to use imino sugars as molecular chaperons that assist protein folding and stability of mutant enzymes. The principles of chaperon-mediated therapy (CMT) are described, and the potential efficacy and preclinical status of imino sugars is compared with substrate reduction therapy (SRT). The increasing use of imino sugars for clinical evaluation of a group of storage diseases that are complex and often intractable disorders to treat has considerable benefit. This is particularly so given the ability of small molecules to be orally available, penetrate the central nervous system (CNS), and have well-characterized biological and pharmacological properties. PMID- 15901677 TI - Pregnancy outcome after blastocyst transfer as compared to early cleavage stage embryo transfer. PMID- 15901678 TI - Preimplantation genetic diagnosis for an insertional translocation. PMID- 15901679 TI - Olfactometric and rhinomanometric outcomes in post-menopausal women treated with hormone therapy: a prospective study. PMID- 15901680 TI - Fertility outcome after a randomized trial comparing curettage with misoprostol for treatment of early pregnancy failure. PMID- 15901681 TI - Layers of signaling in a bacterium-host association. PMID- 15901682 TI - Neutral lipid bodies in prokaryotes: recent insights into structure, formation, and relationship to eukaryotic lipid depots. PMID- 15901683 TI - Vibrio fischeri uses two quorum-sensing systems for the regulation of early and late colonization factors. AB - Vibrio fischeri possesses two quorum-sensing systems, ain and lux, using acyl homoserine lactones as signaling molecules. We have demonstrated previously that the ain system activates luminescence gene expression at lower cell densities than those required for lux system activation and that both systems are essential for persistent colonization of the squid host, Euprymna scolopes. Here, we asked whether the relative contributions of the two systems are also important at different colonization stages. Inactivation of ain, but not lux, quorum-sensing genes delayed initiation of the symbiotic relationship. In addition, our data suggest that lux quorum sensing is not fully active in the early stages of colonization, implying that this system is not required until later in the symbiosis. The V. fischeri luxI mutant does not express detectable light levels in symbiosis yet initiates colonization as well as the wild type, suggesting that ain quorum sensing regulates colonization factors other than luminescence. We used a recently developed V. fischeri microarray to identify genes that are controlled by ain quorum sensing and could be responsible for the initiation defect. We found 30 differentially regulated genes, including the repression of a number of motility genes. Consistent with these data, ain quorum-sensing mutants displayed an altered motility behavior in vitro. Taken together, these data suggest that the sequential activation of these two quorum-sensing systems with increasing cell density allows the specific regulation of early colonization factors (e.g., motility) by ain quorum sensing, whereas late colonization factors (e.g., luminescence) are preferentially regulated by lux quorum sensing. PMID- 15901684 TI - Biosynthetic pathway of Pseudomonas aeruginosa 4-hydroxy-2-alkylquinolines. AB - The role of intercellular communication in the regulation of bacterial multicellular behavior has received widespread attention, and a variety of signal molecules involved in bacterial communication have been discovered. In addition to the N-acyl-homoserine lactones, 4-hydroxy-2-alkylquinolines (HAQs), including the Pseudomonas quinolone signal, have been shown to function as signal molecules in Pseudomonas aeruginosa. In this study we unraveled the biosynthetic pathway of HAQs using feeding experiments with isotope-labeled precursors and analysis of extracted HAQs by gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy. Our results show that the biosynthesis of various HAQ metabolites is directed via a common metabolic pathway involving a "head-to-head" condensation of anthranilic acid and beta-keto fatty acids. Moreover, we provide evidence that the beta-keto-(do)decanoic acids, crucial for the biosynthesis of the heptyl and nonyl derivatives of the 4-hydroxyquinolines in P. aeruginosa, are at least in part derived from a common pool of beta-hydroxy(do)decanoic acids involved in rhamnolipid biosynthesis. PMID- 15901685 TI - The archaeon Pyrococcus horikoshii possesses a bifunctional enzyme for formaldehyde fixation via the ribulose monophosphate pathway. AB - Pyrococcus horikoshii OT3, a hyperthermophilic and anaerobic archaeon, was found to have an open reading frame (PH1938) whose deduced amino acid sequence of the N terminal and C-terminal halves showed significant similarity to two key enzymes of the ribulose monophosphate pathway for formaldehyde fixation in methylotrophic bacteria, 3-hexulose-6-phosphate synthase (HPS) and 6-phospho-3-hexuloisomerase (PHI), respectively. The organism constitutively produced the encoded protein and exhibited activity of the sequential HPS- and PHI-mediated reactions in a particulate fraction. The full-length gene encoding the hybrid enzyme, the sequence corresponding to the HPS region, and the sequence corresponding to the PHI region were expressed in Escherichia coli and were found to produce active enzymes, rHps-Phi, rHps, or rPhi, respectively. Purified rHps-Phi and rHps were found to be active at the growth temperatures of the parent strain, but purified rPhi exhibited significant susceptibility to heat, suggesting that thermostability of the PHI moiety of the bifunctional enzyme (rHps-Phi) resulted from fusion with HPS. The bifunctional enzyme catalyzed the sequential reaction much more efficiently than a mixture of rHps and rPhi. These and other biochemical characterizations of the PH1938 gene product suggest that the ribulose monophosphate pathway plays a significant role in the archaeon under extreme environmental conditions. PMID- 15901686 TI - Recycling of the anhydro-N-acetylmuramic acid derived from cell wall murein involves a two-step conversion to N-acetylglucosamine-phosphate. AB - Escherichia coli breaks down over 60% of the murein of its side wall and reuses the component amino acids to synthesize about 25% of the cell wall for the next generation. The amino sugars of the murein are also efficiently recycled. Here we show that the 1,6-anhydro-N-acetylmuramic acid (anhMurNAc) is returned to the biosynthetic pathway by conversion to N-acetylglucosamine-phosphate (GlcNAc-P). The sugar is first phosphorylated by anhydro-N-acetylmuramic acid kinase (AnmK), yielding MurNAc-P, and this is followed by action of an etherase which cleaves the bond between D-lactic acid and the N-acetylglucosamine moiety of MurNAc-P, yielding GlcNAc-P. The kinase gene has been identified by a reverse genetics method. The enzyme was overexpressed, purified, and characterized. The cell extract of an anmK deletion mutant totally lacked activity on anhMurNAc. Surprisingly, in the anmK mutant, anhMurNAc did not accumulate in the cytoplasm but instead was found in the medium, indicating that there was rapid efflux of free anhMurNAc. PMID- 15901687 TI - Bordetella AlcS transporter functions in alcaligin siderophore export and is central to inducer sensing in positive regulation of alcaligin system gene expression. AB - Bordetella pertussis and Bordetella bronchiseptica, which are respiratory mucosal pathogens of mammals, produce and utilize the siderophore alcaligin to acquire iron in response to iron starvation. A predicted permease of the major facilitator superfamily class of membrane efflux pumps, AlcS (synonyms, OrfX and Bcr), was reported to be encoded within the alcaligin gene cluster. In this study, alcS null mutants were found to be defective in growth under iron starvation conditions, in iron source utilization, and in alcaligin export. trans complementation using cloned alcS genes of B. pertussis or B. bronchiseptica restored the wild-type phenotype to the alcS mutants. Although the levels of extracellular alcaligin measured in alcS strain culture fluids were severely reduced compared with the wild-type levels, alcS mutants had elevated levels of cell-associated alcaligin, implicating AlcS in alcaligin export. Interestingly, a deltaalcA mutation that eliminated alcaligin production suppressed the growth defects of alcS mutants. This suppression and the alcaligin production defect were reversed by trans complementation of the deltaalcA mutation in the double mutant strain, confirming that the growth-defective phenotype of alcS mutants is associated with alcaligin production. In an alcA::mini-Tn5 lacZ1 operon fusion strain background, an alcS null mutation resulted in enhanced AlcR-dependent transcriptional responsiveness to alcaligin inducer; conversely, AlcS overproduction blunted the transcriptional response to alcaligin. These transcription studies indicate that the alcaligin exporter activity of AlcS is required to maintain appropriate intracellular alcaligin levels for normal inducer sensing and responsiveness necessary for positive regulation of alcaligin system gene expression. PMID- 15901688 TI - The Campylobacter jejuni response regulator, CbrR, modulates sodium deoxycholate resistance and chicken colonization. AB - Two-component regulatory systems play a major role in the physiological response of bacteria to environmental stimuli. Such systems are composed of a sensor histidine kinase and a response regulator whose ultimate function is to affect the expression of target genes. Response regulator mutants of Campylobacter jejuni strain F38011 were screened for sensitivity to sodium deoxycholate. A mutation in Cj0643, which encodes a response regulator with no obvious cognate histidine kinase, resulted in an absence of growth on plates containing a subinhibitory concentration of sodium deoxcholate (1%, wt/vol). In broth cultures containing 0.05% (wt/vol) sodium deoxycholate, growth of the mutant was significantly inhibited compared to growth of the C. jejuni F38011 wild-type strain. Complementation of the C. jejuni cbrR mutant in trans restored growth in both broth and plate cultures supplemented with sodium deoxycholate. Based on the phenotype displayed by its mutation, we designated the gene corresponding to Cj0643 as cbrR (Campylobacter bile resistance regulator). While the MICs of a variety of bile salts and other detergents for the C. jejuni cbrR mutant were lower, no difference was noted in its sensitivity to antibiotics or osmolarity. Finally, chicken colonization studies demonstrated that the C. jejuni cbrR mutant had a reduced ability to colonize compared to the wild-type strain. These data support previous findings that bile resistance contributes to colonization of chickens and establish that the response regulator, CbrR, modulates resistance to bile salts in C. jejuni. PMID- 15901689 TI - Characterization of two new aminopeptidases in Escherichia coli. AB - Two genes in the Escherichia coli genome, ypdE and ypdF, have been cloned and expressed, and their products have been purified. YpdF is shown to be a metalloenzyme with Xaa-Pro aminopeptidase activity and limited methionine aminopeptidase activity. Genes homologous to ypdF are widely distributed in bacterial species. The unique feature in the sequences of the products of these genes is a conserved C-terminal domain and a variable N-terminal domain. Full or partial deletion of the N terminus in YpdF leads to the loss of enzymatic activity. The conserved C-terminal domain is homologous to that of the methionyl aminopeptidase (encoded by map) in E. coli. However, YpdF and Map differ in their preference for the amino acid next to the initial methionine in the peptide substrates. The implication of this difference is discussed. ypdE is the immediate downstream gene of ypdF, and its start codon overlaps with the stop codon of ypdF by 1 base. YpdE is shown to be a metalloaminopeptidase and has a broad exoaminopeptidase activity. PMID- 15901690 TI - Levels and activity of the Pseudomonas putida global regulatory protein Crc vary according to growth conditions. AB - The global regulatory protein Crc is involved in the repression of several catabolic pathways for sugars, hydrocarbons, and nitrogenated and aromatic compounds in Pseudomonas putida and Pseudomonas aeruginosa when other preferred carbon sources are present in the culture medium (catabolite repression), therefore modulating carbon metabolism. We have analyzed whether the levels or the activity of Crc is regulated. Crc activity was followed by its ability to inhibit the induction by alkanes of the P. putida OCT plasmid alkane degradation pathway when cells grow in a complete medium, where the effect of Crc is very strong. The abundance of crc transcripts and the amounts of Crc protein were higher under repressing conditions than under nonrepressing conditions. The presence of crc on a high-copy-number plasmid considerably increased Crc levels, but this impaired its ability to inhibit the alkane degradation pathway. Crc shows similarity to a family of nucleases that have highly conserved residues at their catalytic sites. Mutation of the corresponding residues in Crc (Asp220 and His246) led to proteins that can inhibit induction of the alkane degradation pathway when present at normal or elevated levels in the cell. Repression by these mutant proteins occurred only under repressing conditions. These results suggest that both the amounts and the activity of Crc are modulated and support previous proposals that Crc may form part of a signal transduction pathway. Furthermore, the activity of the mutant proteins suggests that Crc is not a nuclease. PMID- 15901691 TI - Iron-responsive regulation of the Helicobacter pylori iron-cofactored superoxide dismutase SodB is mediated by Fur. AB - Maintaining iron homeostasis is a necessity for all living organisms, as free iron augments the generation of reactive oxygen species like superoxide anions, at the risk of subsequent lethal cellular damage. The iron-responsive regulator Fur controls iron metabolism in many bacteria, including the important human pathogen Helicobacter pylori, and thus is directly or indirectly involved in regulation of oxidative stress defense. Here we demonstrate that Fur is a direct regulator of the H. pylori iron-cofactored superoxide dismutase SodB, which is essential for the defense against toxic superoxide radicals. Transcription of the sodB gene was iron induced in H. pylori wild-type strain 26695, resulting in expression of the SodB protein in iron-replete conditions but an absence of expression in iron-restricted conditions. Mutation of the fur gene resulted in constitutive, iron-independent expression of SodB. Recombinant H. pylori Fur protein bound with low affinity to the sodB promoter region, but addition of the iron substitute Mn2+ abolished binding. The operator sequence of the iron-free form of Fur, as identified by DNase I footprinting, was located directly upstream of the sodB gene at positions -5 to -47 from the transcription start site. The direct role of Fur in regulation of the H. pylori sodB gene contrasts with the small-RNA-mediated sodB regulation observed in Escherichia coli. In conclusion, H. pylori Fur is a versatile regulator involved in many pathways essential for gastric colonization, including superoxide stress defense. PMID- 15901692 TI - Characterization of pathways dependent on the uvsE, uvrA1, or uvrA2 gene product for UV resistance in Deinococcus radiodurans. AB - The genome of a radiation-resistant bacterium, Deinococcus radiodurans, contains one uvsE gene and two uvrA genes, uvrA1 and uvrA2. Using a series of mutants lacking these genes, we determined the biological significance of these components to UV resistance. The UV damage endonuclease (UvsE)-dependent excision repair (UVER) pathway and UvrA1-dependent pathway show some redundancy in their function to counteract the lethal effects of UV. Loss of these pathways does not cause increased sensitivity to UV mutagenesis, suggesting either that these pathways play no function in inducing mutations or that there are mechanisms to prevent mutation other than these excision repair pathways. UVER efficiently removes both cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts (6-4PPs) from genomic DNA. In contrast, the UvrA1 pathway does not significantly contribute to the repair of CPDs but eliminates 6-4PPs. Inactivation of uvrA2 does not result in a deleterious effect on survival, mutagenesis, or the repair kinetics of CPDs and 6-4PPs, indicating a minor role in resistance to UV. Loss of uvsE, uvrA1, and uvrA2 reduces but does not completely abolish the ability to eliminate CPDs and 6-4PPs from genomic DNA. The result indicates the existence of a system that removes UV damage yet to be identified. PMID- 15901693 TI - Structural comparison of ten serotypes of staphylocoagulases in Staphylococcus aureus. AB - Staphylocoagulase detection is the hallmark of a Staphylococcus aureus infection. Ten different serotypes of staphylocoagulases have been reported to date. We determined the nucleotide sequences of seven staphylocoagulase genes (coa) and their surrounding regions to compare structures of all 10 staphylocoagulase serotypes, and we inferred their derivations. We found that all staphylocoagulases are comprised of six regions: signal sequence, D1 region, D2 region, central region, repeat region, and C-terminal sequence. Amino acids at both ends, 33 amino acids in the N terminal (the signal sequences and the seven N terminal amino acids in the D1 region) and 5 amino acids in the C terminal, were exactly identical among the 10 serotypes. The central regions were conserved with identities between 80.6 and 94.1% and similarities between 82.8 and 94.6%. Repeat regions comprising tandem repeats of 27 amino acids with a 92% identity on average were polymorphic in the number of repeats. On the other hand, D1 regions other than the seven N-terminal amino acids and D2 regions were less homologous, with diverged identities from 41.5 to 84.5% and 47.0 to 88.9%, respectively, and similarities from 53.5 to 88.7% and 56.8 to 91.9%, respectively, although the predicted prothrombin-binding sites were conserved among them. In contrast, flanking regions of coa were highly homologous, with nucleotide identities of more than 97.1%. Phylogenetic relations among coa did not correlate with those among the flanking regions or housekeeping genes used for multilocus sequence typing. These data indicate that coa could be transmitted to S. aureus, while the less homologous regions in coa presumed to be responsible for different antigenicities might have evolved independently. PMID- 15901694 TI - 16S rRNA mutations that confer tetracycline resistance in Helicobacter pylori decrease drug binding in Escherichia coli ribosomes. AB - Tetracycline resistance in clinical isolates of Helicobacter pylori has been associated with nucleotide substitutions at positions 965 to 967 in the 16S rRNA. We constructed mutants which had different sequences at 965 to 967 in the 16S rRNA gene present on a multicopy plasmid in Escherichia coli strain TA527, in which all seven rrn genes were deleted. The MICs for tetracycline of all mutants having single, double, or triple substitutions at the 965 to 967 region that were previously found in highly resistant H. pylori isolates were higher than that of the mutant exhibiting the wild-type sequence of tetracycline-susceptible H. pylori. The MIC of the mutant with the 965TTC967 triple substitution was 32 times higher than that of the E. coli mutant with the 965AGA967 substitution present in wild-type H. pylori. The ribosomes extracted from the tetracycline-resistant E. coli 965TTC967 variant bound less tetracycline than E. coli with the wild-type H. pylori sequence at this region. The concentration of tetracycline bound to the ribosome was 40% that of the wild type. The results of this study suggest that tetracycline binding to the primary binding site (Tet-1) of the ribosome at positions 965 to 967 is influenced by its sequence patterns, which form the primary binding site for tetracycline. PMID- 15901695 TI - Regulation of mtrF expression in Neisseria gonorrhoeae and its role in high-level antimicrobial resistance. AB - The obligate human pathogen Neisseria gonorrhoeae uses the MtrC-MtrD-MtrE efflux pump to resist structurally diverse hydrophobic antimicrobial agents (HAs), some of which bathe mucosal surfaces that become infected during transmission of gonococci. Constitutive high-level HA resistance occurs by the loss of a repressor (MtrR) that negatively controls transcription of the mtrCDE operon. This high-level HA resistance also requires the product of the mtrF gene, which is located downstream and transcriptionally divergent from mtrCDE. MtrF is a putative inner membrane protein, but its role in HA resistance mediated by the MtrC-MtrD-MtrE efflux pump remains to be determined. High-level HA resistance can also be mediated through an induction process that requires enhanced transcription of mtrCDE when gonococci are grown in the presence of a sublethal concentration of Triton X-100. We now report that inactivation of mtrF results in a significant reduction in the induction of HA resistance and that the expression of mtrF is enhanced when gonococci are grown under inducing conditions. However, no effect was observed on the induction of mtrCDE expression in an MtrF-negative strain. The expression of mtrF was repressed by MtrR, the major repressor of mtrCDE expression. In addition to MtrR, another repressor (MpeR) can downregulate the expression of mtrF. Repression of mtrF by MtrR and MpeR was additive, demonstrating that the repressive effects mediated by these regulators are independent processes. PMID- 15901696 TI - Expression and site-directed mutagenesis of the lactococcal abortive phage infection protein AbiK. AB - Abortive infection mechanisms of Lactococcus lactis form a heterogeneous group of phage resistance systems that act after early phage gene expression. One of these systems, AbiK, aborts infection of the three most prevalent lactococcal phage groups of the dairy industry. In this study, it is demonstrated that the antiphage activity depends on the level of expression of the abiK gene and on the presence of a reverse transcriptase (RT) motif in AbiK. The abiK gene was shown to be part of an operon that includes two additional open reading frames, with one of these encoding a phage-related transcriptional repressor named Orf4. Expression of AbiK is driven by two promoters, PabiK and Porf3, the latter being repressed by Orf4 in vivo. Binding of the purified Orf4 to the Porf3 promoter was demonstrated in vitro by gel retardation assays. The N-terminal half of the deduced AbiK protein possesses an RT motif that was modified by site-directed mutagenesis. Conservative mutations in key positions resulted in the complete loss of the resistance phenotype. These data suggest that an RT activity might be involved in the phage resistance activity of AbiK. A model for the mode of action of AbiK is proposed. PMID- 15901697 TI - Purification and characterization of allophanate hydrolase (AtzF) from Pseudomonas sp. strain ADP. AB - AtzF, allophanate hydrolase, is a recently discovered member of the amidase signature family that catalyzes the terminal reaction during metabolism of s triazine ring compounds by bacteria. In the present study, the atzF gene from Pseudomonas sp. strain ADP was cloned and expressed as a His-tagged protein, and the protein was purified and characterized. AtzF had a deduced subunit molecular mass of 66,223, based on the gene sequence, and an estimated holoenzyme molecular mass of 260,000. The active protein did not contain detectable metals or organic cofactors. Purified AtzF hydrolyzed allophanate with a k(cat)/K(m) of 1.1 x 10(4) s(-1) M(-1), and 2 mol of ammonia was released per mol allophanate. The substrate range of AtzF was very narrow. Urea, biuret, hydroxyurea, methylcarbamate, and other structurally analogous compounds were not substrates for AtzF. Only malonamate, which strongly inhibited allophanate hydrolysis, was an alternative substrate, with a greatly reduced k(cat)/K(m) of 21 s(-1) M(-1). Data suggested that the AtzF catalytic cycle proceeds through a covalent substrate-enzyme intermediate. AtzF reacts with malonamate and hydroxylamine to generate malonohydroxamate, potentially derived from hydroxylamine capture of an enzyme tethered acyl group. Three putative catalytically important residues, one lysine and two serines, were altered by site-directed mutagenesis, each with complete loss of enzyme activity. The identity of a putative serine nucleophile was probed using phenyl phosphorodiamidate that was shown to be a time-dependent inhibitor of AtzF. Inhibition was due to phosphoroamidation of Ser189 as shown by liquid chromatography/matrix-assisted laser desorption ionization mass spectrometry. The modified residue corresponds in sequence alignments to the nucleophilic serine previously identified in other members of the amidase signature family. Thus, AtzF affects the cleavage of three carbon-to-nitrogen bonds via a mechanism similar to that of enzymes catalyzing single-amide-bond cleavage reactions. AtzF orthologs appear to be widespread among bacteria. PMID- 15901698 TI - Novel molecular features of the fibrolytic intestinal bacterium Fibrobacter intestinalis not shared with Fibrobacter succinogenes as determined by suppressive subtractive hybridization. AB - Suppressive subtractive hybridization was conducted to identify unique genes coding for plant cell wall hydrolytic enzymes and other properties of the gastrointestinal bacterium Fibrobacter intestinalis DR7 not shared by Fibrobacter succinogenes S85. Subtractive clones from F. intestinalis were sequenced and assembled to form 712 nonredundant contigs with an average length of 525 bp. Of these, 55 sequences were unique to F. intestinalis. The remaining contigs contained 764 genes with BLASTX similarities to other proteins; of these, 80% had the highest similarities to proteins in F. succinogenes, including 30 that coded for carbohydrate active enzymes. The expression of 17 of these genes was verified by Northern dot blot analysis. Of genes not exhibiting BLASTX similarity to F. succinogenes, 30 encoded putative transposases, 6 encoded restriction modification genes, and 45% had highest similarities to proteins in other species of gastrointestinal bacteria, a finding suggestive of either horizontal gene transfer to F. intestinalis or gene loss from F. succinogenes. Analysis of contigs containing segments of two or more adjacent genes revealed that only 35% exhibited BLASTX similarity and were in the same orientation as those of F. succinogenes, indicating extensive chromosomal rearrangement. The expression of eight transposases, and three restriction-modification genes was confirmed by Northern dot blot analysis. These data clearly document the maintenance of carbohydrate active enzymes in F. intestinalis necessitated by the preponderance of polysaccharide substrates available in the ruminal environment. It also documents substantive changes in the genome from that of F. succinogenes, which may be related to the introduction of the array of transposase and restriction modification genes. PMID- 15901699 TI - Fructose utilization in Lactococcus lactis as a model for low-GC gram-positive bacteria: its regulator, signal, and DNA-binding site. AB - In addition to its role as carbon and energy source, fructose metabolism was reported to affect other cellular processes, such as biofilm formation by streptococci and bacterial pathogenicity in plants. Fructose genes encoding a 1 phosphofructokinase and a phosphotransferase system (PTS) fructose-specific enzyme IIABC component reside commonly in a gene cluster with a DeoR family regulator in various gram-positive bacteria. We present a comprehensive study of fructose metabolism in Lactococcus lactis, including a systematic study of fru mutants, global messenger analysis, and a molecular characterization of its regulation. The fru operon is regulated at the transcriptional level by both FruR and CcpA and at the metabolic level by inducer exclusion. The FruR effector is fructose-1-phosphate (F1P), as shown by combined analysis of transcription and measurements of the intracellular F1P pools in mutants either unable to produce this metabolite or accumulating it. The regulation of the fru operon by FruR requires four adjacent 10-bp direct repeats. The well-conserved organization of the fru promoter region in various low-GC gram-positive bacteria, including CRE boxes as well as the newly defined FruR motif, suggests that the regulation scheme defined in L. lactis could be applied to these bacteria. Transcriptome profiling of fruR and fruC mutants revealed that the effect of F1P and FruR regulation is limited to the fru operon in L. lactis. This result is enforced by the fact that no other targets for FruR were found in the available low-GC gram positive bacteria genomes, suggesting that additional phenotypical effects due to fructose metabolism do not rely directly on FruR control, but rather on metabolism. PMID- 15901700 TI - Sulfur amino acid metabolism and its control in Lactococcus lactis IL1403. AB - Cysteine and methionine availability influences many processes in the cell. In bacteria, transcription of the specific genes involved in the synthesis of these two amino acids is usually regulated by different mechanisms or regulators. Pathways for the synthesis of cysteine and methionine and their interconversion were experimentally determined for Lactococcus lactis, a lactic acid bacterium commonly found in food. A new gene, yhcE, was shown to be involved in methionine recycling to cysteine. Surprisingly, 18 genes, representing almost all genes of these pathways, are under the control of a LysR-type activator, FhuR, also named CmbR. DNA microarray experiments showed that FhuR targets are restricted to this set of 18 genes clustered in seven transcriptional units, while cysteine starvation modifies the transcription level of several other genes potentially involved in oxidoreduction processes. Purified FhuR binds a 13-bp box centered 46 to 53 bp upstream of the transcriptional starts from the seven regulated promoters, while a second box with the same consensus is present upstream of the first binding box, separated by 8 to 10 bp. O-Acetyl serine increases FhuR binding affinity to its binding boxes. The overall view of sulfur amino acid metabolism and its regulation in L. lactis indicates that CysE could be a master enzyme controlling the activity of FhuR by providing its effector, while other controls at the enzymatic level appear to be necessary to compensate the absence of differential regulation of the genes involved in the interconversion of methionine and cysteine and other biosynthesis genes. PMID- 15901701 TI - Role of pi dimers in coupling ("handcuffing") of plasmid R6K's gamma ori iterons. AB - One proposed mechanism of replication inhibition in iteron-containing plasmids (ICPs) is "handcuffing," in which the coupling of origins via iteron-bound replication initiator (Rep) protein turns off origin function. In minimal R6K replicons, copy number control requires the interaction of plasmid-encoded pi protein with the seven 22-bp iterons of the gamma origin of replication. Like other related Rep proteins, pi exists as both monomers and dimers. However, the ability of pi dimers to bind iterons distinguishes R6K from most other ICPs, where only monomers have been observed to bind iterons. Here, we describe experiments to determine if monomers or dimers of pi protein are involved in the formation of handcuffed complexes. Standard ligation enhancement assays were done using pi variants with different propensities to bind iterons as monomers or dimers. Consistent with observations from several ICPs, a hyperreplicative variant (pi.P106L(wedge)F107S) exhibits deficiencies in handcuffing. Additionally, a novel dimer-biased variant of pi protein (pi.M36A(wedge)M38A), which lacks initiator function, handcuffs iteron-containing DNA more efficiently than does wild-type pi. The data suggest that pi dimers mediate handcuffing, supporting our previously proposed model of handcuffing in the gamma ori system. Thus, dimers of pi appear to possess three distinct inhibitory functions with respect to R6K replication: transcriptional autorepression of pi expression, in cis competition (for origin binding) with monomeric activator pi, and handcuffing mediated inhibition of replication in trans. PMID- 15901702 TI - Characterization of methionine export in Corynebacterium glutamicum. AB - Corynebacterium glutamicum is known for its effective excretion of amino acids under particular metabolic conditions. Concomitant activities of uptake and excretion systems would create an energy-wasting futile cycle; amino acid export systems are therefore tightly regulated. We have used a DNA microarray approach to identify genes for membrane proteins which are overexpressed under conditions of elevated cytoplasmic concentrations of methionine. One of these genes was brnF, coding for the larger subunit of BrnFE, a previously identified two component isoleucine export system. By deletion, complementation, and overexpression of the brnFE genes in a C. glutamicum strain, in which the two uptake systems for methionine were inactivated, we identified BrnFE as being responsible for methionine export. In the presence of both substrates in the cytoplasm, BrnFE was found to transport isoleucine and methionine at similar rates. The expression of the brnFE gene cluster depends on an Lrp-type transcription factor and was shown to be strongly induced by increasing cytoplasmic methionine concentration. Methionine was a better inducer than isoleucine, indicating that methionine rather than isoleucine might be the native substrate of BrnFE. When the synthesis of BrnFE was blocked by chloramphenicol, fast methionine export was still observed, but only at greatly increased cytoplasmic levels of this amino acid. This indicates the presence of at least one other methionine export system, presumably with low affinity but high capacity. Under conditions where cytoplasmic methionine does not exceed a concentration of 50 mM, BrnFE is the dominant export system for this amino acid. PMID- 15901703 TI - Alteration of the fatty acid profile of Streptomyces coelicolor by replacement of the initiation enzyme 3-ketoacyl acyl carrier protein synthase III (FabH). AB - The first elongation step of fatty acid biosynthesis by a type II dissociated fatty acid synthases is catalyzed by 3-ketoacyl-acyl carrier protein (ACP) synthase III (KASIII, FabH). This enzyme, encoded by the fabH gene, catalyzes a decarboxylative condensation between an acyl coenzyme A (CoA) primer and malonyl ACP. In organisms such as Escherichia coli, which generate only straight-chain fatty acids (SCFAs), FabH has a substrate preference for acetyl-CoA. In streptomycetes and other organisms which produce a mixture of both SCFAs and branched-chain fatty acids (BCFAs), FabH has been shown to utilize straight- and branched-chain acyl-CoA substrates. We report herein the generation of a Streptomyces coelicolor mutant (YL/ecFabH) in which the chromosomal copy of the fabH gene has been replaced and the essential process of fatty acid biosynthesis is initiated by plasmid-based expression of the E. coli FabH (bearing only 35% amino acid identity to the Streptomyces enzyme). The YL/ecFabH mutant produces predominantly SCFAs (86%). In contrast, BCFAs predominate (approximately 70%) in both the S. coelicolor parental strain and S. coelicolor YL/sgFabH (a deltafabH mutant carrying a plasmid expressing the Streptomyces glaucescens FabH). These results provide the first unequivocal evidence that the substrate specificity of FabH observed in vitro is a determinant of the fatty acid made in an organism. The YL/ecFabH strain grows significantly slower on both solid and liquid media. The levels of FabH activity in cell extracts of YL/ecFabH were also significantly lower than those in cell extracts of YL/sgFabH, suggesting that a decreased rate of fatty acid synthesis may account for the observed decreased growth rate. The production of low levels of BCFAs in YL/ecFabH suggests either that the E. coli FabH is more tolerant of different acyl-CoAs substrates than previously thought or that there is an additional pathway for initiation of BCFA biosynthesis in Streptomyces coelicolor. PMID- 15901704 TI - The ExsA protein of Bacillus cereus is required for assembly of coat and exosporium onto the spore surface. AB - The outermost layer of spores of the Bacillus cereus family is a loose structure known as the exosporium. Spores of a library of Tn917-LTV1 transposon insertion mutants of B. cereus ATCC 10876 were partitioned into hexadecane; a less hydrophobic mutant that was isolated contained an insertion in the exsA promoter region. ExsA is the equivalent of SafA (YrbA) of Bacillus subtilis, which is also implicated in spore coat assembly; the gene organizations around both are identical, and both proteins contain a very conserved N-terminal cortex-binding domain of ca. 50 residues, although the rest of the sequence is much less conserved. In particular, unlike SafA, the ExsA protein contains multiple tandem oligopeptide repeats and is therefore likely to have an extended structure. The exsA gene is expressed in the mother cell during sporulation. Spores of an exsA mutant are extremely permeable to lysozyme and are blocked in late stages of germination, which require coat-associated functions. Two mutants expressing differently truncated versions of ExsA were constructed, and they showed the same gross defects in the attachment of exosporium and spore coat layers. The protein profile of the residual exosporium harvested from spores of the three mutants- two expressing truncated proteins and the mutant with the original transposon insertion in the promoter region--showed some differences from the wild type and from each other, but the major exosporium glycoproteins were retained. The exsA gene is extremely important for the normal assembly and anchoring of both the spore coat and exosporium layers in spores of B. cereus. PMID- 15901705 TI - Identification of a turnover element in region 2.1 of Escherichia coli sigma32 by a bacterial one-hybrid approach. AB - Induction of the heat shock response in Escherichia coli requires the alternative sigma factor sigma32 (RpoH). The cellular concentration of sigma32 is controlled by proteolysis involving FtsH, other proteases, and the DnaKJ chaperone system. To identify individual sigma32 residues critical for degradation, we used a recently developed bacterial one-hybrid system and screened for stabilized versions of sigma32. The five single point mutations that rendered the sigma factor more stable mapped to positions L47, A50, and I54 in region 2.1. Strains expressing the stabilized sigma32 variants exhibited elevated transcriptional activity, as determined by a groE-lacZ fusion. Structure calculations predicted that the three mutated residues line up on the same face of an alpha-helix in region 2.1, suggesting that they are positioned to interact with proteins of the degradation machinery. PMID- 15901706 TI - Kinetic studies of polyhydroxybutyrate granule formation in Wautersia eutropha H16 by transmission electron microscopy. AB - Wautersia eutropha, formerly known as Ralstonia eutropha, a gram-negative bacterium, accumulates polyhydroxybutyrate (PHB) as insoluble granules inside the cell when nutrients other than carbon are limited. In this paper, we report findings from kinetic studies of granule formation and degradation in W. eutropha H16 obtained using transmission electron microscopy (TEM). In nitrogen-limited growth medium, the phenotype of the cells at the early stages of granule formation was revealed for the first time. At the center of the cells, dark stained "mediation elements" with small granules attached were observed. These mediation elements are proposed to serve as nucleation sites for granule initiation. TEM images also revealed that when W. eutropha cells were introduced into nitrogen-limited medium from nutrient-rich medium, the cell size increased two- to threefold, and the cells underwent additional volume changes during growth. Unbiased stereology was used to analyze the two-dimensional TEM images, from which the average volume of a W. eutropha H16 cell and the total surface area of granules per cell in nutrient-rich and PHB production media were obtained. These parameters were essential in the calculation of the concentration of proteins involved in PHB formation and utilization and their changes with time. The extent of protein coverage of the granule surface area is presented in the accompanying paper. PMID- 15901707 TI - Analysis of transient polyhydroxybutyrate production in Wautersia eutropha H16 by quantitative Western analysis and transmission electron microscopy. AB - Polyhydroxybutyrates (PHBs) are polyoxoesters generated from (R)3-hydroxybutyryl coenzyme A by PHB synthase. During the polymerization reaction, the polymers undergo a phase transition and generate granules. Wautersia eutropha can transiently accumulate PHB when it is grown in a nutrient-rich medium (up to 23% of the cell dry weight in dextrose-free tryptic soy broth [TSB]). PHB homeostasis under these growth conditions was examined by quantitative Western analysis to monitor the proteins present, their levels, and changes in their levels over a 48 h growth period. The proteins examined include PhaC (the synthase), PhaP (a phasin), PhaR (a transcription factor), and PhaZ1(a), PhaZ1(b), and PhaZ1(c) (putative intracellular depolymerases), as well as PhaZ2 (a hydroxybutyrate oligomer hydrolase). The results show that PhaC and PhaZ1(a) were present simultaneously. No PhaZ1(b) or PhaZ1(c) was detected at any time throughout growth. PhaZ2 was observed and exhibited an expression pattern different from that of PhaZ1(a). The levels of PhaP changed dramatically and corresponded kinetically to the levels of PHB. Transmission electron microscopy (TEM) provided the dimensions of the average cell and the average granule at 4 h and 24 h of growth (J. Tian, A. J. Sinskey, and J. Stubbe, J. Bacteriol. 187:3814-3824, 2005). This information allowed us to calculate the amount of each protein and number of granules per cell and the granule surface coverage by proteins. The molecular mass of PHB (10(6) Da) was determined by dynamic light scattering at 4 h, the time of maximum PHB accumulation. At this time, the surface area of the granules was maximally covered with PhaP (27 to 54%), and there were one or two PhaP molecules/PHB chain. The ratio of PHB chains to PhaC was approximately 60, which required reinitiation of polymer formation on PhaC. The TEM studies of wild type and deltaphaR strains in TSB provided further support for an alternative mechanism of granule formation. PMID- 15901708 TI - Structure-based site-directed mutagenesis of the UDP-MurNAc-pentapeptide-binding cavity of the FemX alanyl transferase from Weissella viridescens. AB - Weissella viridescens FemX (FemX(Wv)) belongs to the Fem family of nonribosomal peptidyl transferases that use aminoacyl-tRNA as the amino acid donor to synthesize the peptide cross-bridge found in the peptidoglycan of many species of pathogenic gram-positive bacteria. We have recently solved the crystal structure of FemX(Wv) in complex with the peptidoglycan precursor UDP-MurNAc-pentapeptide and report here the site-directed mutagenesis of nine residues located in the binding cavity for this substrate. Two substitutions, Lys36Met and Arg211Met, depressed FemX(Wv) transferase activity below detectable levels without affecting protein folding. Analogues of UDP-MurNAc-pentapeptide lacking the phosphate groups or the C-terminal D-alanyl residues were not substrates of the enzyme. These results indicate that Lys36 and Arg211 participate in a complex hydrogen bond network that connects the C-terminal D-Ala residues to the phosphate groups of UDP-MurNAc-pentapeptide and constrains the substrate in a conformation that is essential for transferase activity. PMID- 15901709 TI - A multidomain fusion protein in Listeria monocytogenes catalyzes the two primary activities for glutathione biosynthesis. AB - Glutathione is the predominant low-molecular-weight peptide thiol present in living organisms and plays a key role in protecting cells against oxygen toxicity. Until now, glutathione synthesis was thought to occur solely through the consecutive action of two physically separate enzymes, gamma-glutamylcysteine ligase and glutathione synthetase. In this report we demonstrate that Listeria monocytogenes contains a novel multidomain protein (termed GshF) that carries out complete synthesis of glutathione. Evidence for this comes from experiments which showed that in vitro recombinant GshF directs the formation of glutathione from its constituent amino acids and the in vivo effect of a mutation in GshF that abolishes glutathione synthesis, results in accumulation of the intermediate gamma-glutamylcysteine, and causes hypersensitivity to oxidative agents. We identified GshF orthologs, consisting of a gamma-glutamylcysteine ligase (GshA) domain fused to an ATP-grasp domain, in 20 gram-positive and gram-negative bacteria. Remarkably, 95% of these bacteria are mammalian pathogens. A plausible origin for GshF-dependent glutathione biosynthesis in these bacteria was the recruitment by a GshA ancestor gene of an ATP-grasp gene and the subsequent spread of the fusion gene between mammalian hosts, most likely by horizontal gene transfer. PMID- 15901710 TI - Structures of the iron-sulfur flavoproteins from Methanosarcina thermophila and Archaeoglobus fulgidus. AB - Iron-sulfur flavoproteins (ISF) constitute a widespread family of redox-active proteins in anaerobic prokaryotes. Based on sequence homologies, their overall structure is expected to be similar to that of flavodoxins, but in addition to a flavin mononucleotide cofactor they also contain a cubane-type [4Fe:4S] cluster. In order to gain further insight into the function and properties of ISF, the three-dimensional structures of two ISF homologs, one from the thermophilic methanogen Methanosarcina thermophila and one from the hyperthermophilic sulfate reducing archaeon Archaeoglobus fulgidus, were determined. The structures indicate that ISF assembles to form a tetramer and that electron transfer between the two types of redox cofactors requires oligomerization to juxtapose the flavin mononucleotide and [4Fe:4S] cluster bound to different subunits. This is only possible between different monomers upon oligomerization. Fundamental differences in the surface properties of the two ISF homologs underscore the diversity encountered within this protein family. PMID- 15901711 TI - Structure and genome organization of AFV2, a novel archaeal lipothrixvirus with unusual terminal and core structures. AB - A novel filamentous virus, AFV2, from the hyperthermophilic archaeal genus Acidianus shows structural similarity to lipothrixviruses but differs from them in its unusual terminal and core structures. The double-stranded DNA genome contains 31,787 bp and carries eight open reading frames homologous to those of other lipothrixviruses, a single tRNA(Lys) gene containing a 12-bp archaeal intron, and a 1,008-bp repeat-rich region near the center of the genome. PMID- 15901712 TI - Structure-activity analysis of microcinJ25: distinct parts of the threaded lasso molecule are responsible for interaction with bacterial RNA polymerase. AB - Peptide microcin J25 (MccJ25) inhibits bacterial RNA polymerase. We show that thermolysin-cleaved MccJ25 and MccJ25 lacking amino acids 13 to 17 also inhibit transcription. Our data and structural analysis of intact and thermolysin digested MccJ25 suggest that distinct regions of MccJ25 are involved in transcription inhibition and cell entry. PMID- 15901713 TI - Surface structure and nanomechanical properties of Shewanella putrefaciens bacteria at two pH values (4 and 10) determined by atomic force microscopy. AB - The nanomechanical properties of gram-negative bacteria (Shewanella putrefaciens) were investigated in situ in aqueous solutions at two pH values, specifically, 4 and 10, by atomic force microscopy (AFM). For both pH values, the approach force curves exhibited subsequent nonlinear and linear regimens that were related to the progressive indentation of the AFM tip in the bacterial cell wall, including a priori polymeric fringe (nonlinear part), while the linear part was ascribed to compression of the plasma membrane. These results indicate the dynamic of surface ultrastructure in response to changes in pH, leading to variations in nanomechanical properties, such as the Young's modulus and the bacterial spring constant. PMID- 15901714 TI - Alginate lyase (AlgL) activity is required for alginate biosynthesis in Pseudomonas aeruginosa. AB - To determine whether AlgL's lyase activity is required for alginate production in Pseudomonas aeruginosa, an algLdelta::Gm(r) mutant (FRD-MA7) was created. algL complementation of FRD-MA7 restored alginate production, but algL constructs containing mutations inactivating lyase activity did not, demonstrating that the enzymatic activity of AlgL is required for alginate production. PMID- 15901715 TI - Osmotic regulation of the Escherichia coli bdm (biofilm-dependent modulation) gene by the RcsCDB His-Asp phosphorelay. AB - The RcsCDB His-Asp phosphorelay is shown to positively regulate the bdm (biofilm dependent modulation) and sra (stationary-phase-induced ribosome-associated protein) genes in Escherichia coli. The regulation is direct and requires an RcsB box next to the bdm -35 element. In addition, bdm is shown to be activated by osmotic shock in an Rcs-dependent way. PMID- 15901716 TI - Two additional components of the accessory sec system mediating export of the Streptococcus gordonii platelet-binding protein GspB. AB - The gspB-secY2A2 locus of Streptococcus gordonii strain M99 encodes the platelet binding glycoprotein GspB, along with proteins that mediate its glycosylation and export. We have identified two additional components of the accessory Sec system (Asp4 and Asp5) encoded just downstream of gtfB in the gspB-secY2A2 locus. These proteins are required for GspB export and for normal levels of platelet binding by M99. Asp4 and Asp5 may be functional homologues of SecE and SecG, respectively. PMID- 15901717 TI - Methylotrophic autotrophy in Beijerinckia mobilis. AB - Representatives of the genus Beijerinckia are known as heterotrophic, dinitrogen fixing bacteria which utilize a wide range of multicarbon compounds. Here we show that at least one of the currently known species of this genus, i.e., Beijerinckia mobilis, is also capable of methylotrophic metabolism coupled with the ribulose bisphosphate (RuBP) pathway of C1 assimilation. A complete suite of dehydrogenases commonly involved in the sequential oxidation of methanol via formaldehyde and formate to CO2 was detected in cell extracts of B. mobilis grown on CH3OH. Carbon dioxide produced by oxidation of methanol was further assimilated via the RuBP pathway as evidenced by reasonably high activities of phosphoribulokinase and ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO). Detection and partial sequence analysis of genes encoding the large subunits of methanol dehydrogenase (mxaF) and form I RubisCO (cbbL) provided genotypic evidence for methylotrophic autotrophy in B. mobilis. PMID- 15901718 TI - A gene encoding L-methionine gamma-lyase is present in Enterobacteriaceae family genomes: identification and characterization of Citrobacter freundii L-methionine gamma-lyase. AB - Citrobacter freundii cells produce L-methionine gamma-lyase when grown on a medium containing L-methionine. The nucleotide sequence of the hybrid plasmid with a C. freundii EcoRI insert of about 3.0 kbp contained two open reading frames, consisting of 1,194 nucleotides and 1,296 nucleotides, respectively. The first one (denoted megL) encoded L-methionine gamma-lyase. The enzyme was overexpressed in Escherichia coli and purified. The second frame encoded a protein belonging to the family of permeases. Regions of high sequence identity with the 3'-terminal part of the C. freundii megL gene located in the same regions of Salmonella enterica serovar Typhimurium, Shigella flexneri, E. coli, and Citrobacter rodentium genomes were found. PMID- 15901719 TI - The eefABC multidrug efflux pump operon is repressed by H-NS in Enterobacter aerogenes. AB - The Enterobacter aerogenes eefABC locus, which encodes a tripartite efflux pump, was cloned by complementation of an Escherichia coli tolC mutant. E. aerogenes deltaacrA expressing EefABC became less susceptible to a wide range of antibiotics. Data from eef::lacZ fusions showed that eefABC was not transcribed in the various laboratory conditions tested. However, increased transcription from Peef was observed in an E. coli hns mutant. In addition, EefA was detected in E. aerogenes expressing a dominant negative E. coli hns allele. PMID- 15901720 TI - Quorum sensing negatively controls type III secretion regulon expression in Pseudomonas aeruginosa PAO1. AB - A systematic analysis of the type III secretion (T3S) genes of Pseudomonas aeruginosa strain PAO1 revealed that they are under quorum-sensing control. This observation was supported by the down-regulation of the T3S regulon in the presence of RhlR-C4HSL and the corresponding advanced secretion of ExoS in a rhlI mutant. PMID- 15901721 TI - Evolution of subspecies of Francisella tularensis. AB - Analysis of unidirectional genomic deletion events and single nucleotide variations suggested that the four subspecies of Francisella tularensis have evolved by vertical descent. The analysis indicated an evolutionary scenario where the highly virulent F. tularensis subsp. tularensis (type A) appeared before the less virulent F. tularensis subsp. holarctica (type B). Compared to their virulent progenitors, attenuated strains of F. tularensis exhibited specific unidirectional gene losses. PMID- 15901722 TI - Critical hydrophobic interactions between phosphorylation and actuator domains of Ca2+-ATPase for hydrolysis of phosphorylated intermediate. AB - Functional roles of seven hydrophobic residues on the interface between the actuator (A) and phosphorylation (P) domains of sarcoplasmic reticulum Ca2+ ATPase were explored by alanine and serine substitutions. The residues examined were Ile179/Leu180/Ile232 on the A domain, Val705/Val726 on the P domain, and Leu119/Tyr122 on the loop linking the A domain and M2 (the second transmembrane helix). These residues gather to form a hydrophobic cluster around Tyr122 in the crystal structures of Ca2+-ATPase in Ca2+-unbound E2 (unphosphorylated) and E2P (phosphorylated) states but are far apart in those of Ca2+-bound E1 (unphosphorylated) and E1P (phosphorylated) states. The substitution-effects were also compared with those of Ile235 on the A domain/M3 linker and those of T181GE of the A domain, since they are in the immediate vicinity of the Tyr122-cluster. All these substitutions almost completely inhibited ATPase activity without inhibiting Ca2+-activated E1P formation from ATP. Substitutions of Ile235 and T181GE blocked the E1P to E2P transition, whereas those in the Tyr122-cluster blocked the subsequent E2P hydrolysis. Substitutions of Ile235 and Glu183 also blocked EP hydrolysis. Results indicate that the Tyr122-cluster is formed during the E1P to E2P transition to configure the catalytic site and position Glu183 properly for hydrolyzing the acylphosphate. Ile235 on the A domain/M3 linker likely forms hydrophobic interactions with the A domain and thereby allowing the strain of this linker to be utilized for large motions of the A domain during these processes. The Tyr122-cluster, Ile235, and T181GE thus seem to have different roles and are critical in the successive events in processing phosphorylated intermediates to transport Ca2+. PMID- 15901723 TI - NAD+-dependent DNA Ligase (Rv3014c) from Mycobacterium tuberculosis. Crystal structure of the adenylation domain and identification of novel inhibitors. AB - DNA ligases utilize either ATP or NAD+ as cofactors to catalyze the formation of phosphodiester bonds in nicked DNA. Those utilizing NAD+ are attractive drug targets because of the unique cofactor requirement for ligase activity. We report here the crystal structure of the adenylation domain of the Mycobacterium tuberculosis NAD+-dependent ligase with bound AMP. The adenosine nucleoside moiety of AMP adopts a syn-conformation. The structure also captures a new spatial disposition between the two subdomains of the adenylation domain. Based on the crystal structure and an in-house compound library, we have identified a novel class of inhibitors for the enzyme using in silico docking calculations. The glycosyl ureide-based inhibitors were able to distinguish between NAD+- and ATP-dependent ligases as evidenced by in vitro assays using T4 ligase and human DNA ligase I. Moreover, assays involving an Escherichia coli strain harboring a temperature-sensitive ligase mutant and a ligase-deficient Salmonella typhimurium strain suggested that the bactericidal activity of the inhibitors is due to inhibition of the essential ligase enzyme. The results can be used as the basis for rational design of novel antibacterial agents. PMID- 15901724 TI - Formation of an ATP-DnaA-specific initiation complex requires DnaA Arginine 285, a conserved motif in the AAA+ protein family. AB - Escherichia coli DnaA protein, a member of the AAA+ superfamily, initiates replication from the chromosomal origin oriC in an ATP-dependent manner. Nucleoprotein complex formed on oriC with the ATP-DnaA multimer but not the ADP DnaA multimer is competent to unwind the oriC duplex. The oriC region contains ATP-DnaA-specific binding sites termed I2 and I3, which stimulate ATP-DnaA dependent oriC unwinding. In this study, we show that the DnaA R285A mutant is inactive for oriC replication in vivo and in vitro and that the mutation is associated with specific defects in oriC unwinding. In contrast, activities of DnaA R285A are sustained in binding to the typical DnaA boxes and to ATP and ADP, formation of multimeric complexes on oriC, and loading of the DnaB helicase onto single-stranded DNA. Footprint analysis of the DnaA-oriC complex reveals that the ATP form of DnaA R285A does not interact with ATP-DnaA-specific binding sites such as the I sites. A subgroup of DnaA molecules in the oriC complex must contain the Arg-285 residue for initiation. Sequence and structural analyses suggest that the DnaA Arg-285 residue is an arginine finger, an AAA+ family specific motif that recognizes ATP bound to an adjacent subunit in a multimeric complex. In the context of these and previous results, the DnaA Arg-285 residue is proposed to play a unique role in the ATP-dependent conformational activation of an initial complex by recognizing ATP bound to DnaA and by modulating the structure of the DnaA multimer to allow interaction with ATP-DnaA-specific binding sites in the complex. PMID- 15901725 TI - The hydrophobic hinge region of rat DNA polymerase beta is critical for substrate binding pocket geometry. AB - The hydrophobic hinge of DNA polymerase beta facilitates closing and stabilization of the enzyme once the nucleotide substrate has bound. Alteration of the hydrophobic nature of the hinge by the introduction of a hydrophilic glutamine residue in place of isoleucine 260 results in an inaccurate polymerase. The kinetic basis of infidelity is lack of discrimination during the binding of substrate. The I260Q polymerase beta variant has lower affinity than wild type enzyme for the correct substrate and much higher affinity for the incorrect substrate. Our results demonstrate that the hinge is important for formation of the substrate binding pocket. Our results are also consistent with the interpretation that DNA polymerase beta discriminates the correct from incorrect substrate during the binding step. PMID- 15901726 TI - Nuclear import and export signals in control of Nrf2. AB - Nrf2 binds to the antioxidant response element and regulates expression and antioxidant induction of a battery of chemopreventive genes. In this study, we have identified nuclear import and export signals of Nrf2 and show that the nuclear import and export of Nrf2 is regulated by antioxidants. We demonstrate that Nrf2 contains a bipartite nuclear localization signal (NLS) and a leucine rich nuclear export signal, which regulate Nrf2 shuttling in and out of the nucleus. Immunofluorescence and immunoblot analysis revealed that Nrf2 accumulates in the nucleus within 15 min of antioxidant treatment and is exported out of nucleus by 8 h after treatment. Nrf2 mutant lacking the NLS failed to enter the nucleus and displayed diminished expression and induction of the downstream NAD(P)H:quinone oxidoreductase 1 gene. The Nrf2 NLS sequence, when fused to green fluorescence protein, resulted in the nuclear accumulation of green fluorescence protein, indicating that this signal sequence was sufficient to direct nuclear localization of Nrf2. A nuclear export signal (NES) was characterized in the C terminus of Nrf2, the deletion of which caused Nrf2 to accumulate predominantly in the nucleus. The Nrf2 NES was sensitive to leptomycin B and could function as an independent export signal when fused to a heterologous protein. Further studies demonstrate that NES-mediated nuclear export of Nrf2 is required for degradation of Nrf2 in the cytosol. These results led to the conclusion that Nrf2 localization between cytosol and nucleus is controlled by both nuclear import and export of Nrf2, and the overall distribution of Nrf2 is probably the result from a balance between these two processes. Antioxidants change this balance in favor of nuclear accumulation of Nrf2, leading to activation of chemopreventive proteins. Once this is achieved, Nrf2 exits the nucleus for binding to INrf2 and degradation. PMID- 15901727 TI - Analysis of the heteromeric CsdA-CsdE cysteine desulfurase, assisting Fe-S cluster biogenesis in Escherichia coli. AB - Biogenesis of iron-sulfur (Fe-S) cluster-containing proteins relies on assistance of complex machineries. To date three systems, NIF, ISC, and SUF, were reported to allow maturation of Fe-S proteins. Here we report that the csdA-csdE (formally ygdK) genes of Escherichia coli constitute a sulfur-generating system referred to as CSD which also contributes to Fe-S biogenesis in vivo. This conclusion was reached by applying a thorough combination of both in vivo and in vitro strategies and techniques. Yeast two-hybrid analysis allowed us to show that CsdA and CsdE interact. Enzymology analysis showed that CsdA cysteine desulfurase activity is increased 2-fold in the presence of CsdE. Mass spectrometry analysis and site-directed mutagenesis showed that residue Cys-61 from CsdE acted as an acceptor site for sulfur provided by cysteine desulfurase activity of CsdA. Genetic investigations revealed that the csdA-csdE genes could act as multicopy suppressors of iscS mutation. Moreover, both in vitro and in vivo investigations pointed to a specific connection between the CSD system and quinolinate synthetase NadA. PMID- 15901728 TI - Tissue- and gene-specific recruitment of steroid receptor coactivator-3 by thyroid hormone receptor during development. AB - Numerous coactivators that bind nuclear hormone receptors have been isolated and characterized in vitro. Relatively few studies have addressed the developmental roles of these cofactors in vivo. By using the total dependence of amphibian metamorphosis on thyroid hormone (T3) as a model, we have investigated the role of steroid receptor coactivator 3 (SRC3) in gene activation by thyroid hormone receptor (TR) in vivo. First, expression analysis showed that SRC3 was expressed in all tadpole organs analyzed. In addition, during natural as well as T3-induced metamorphosis, SRC3 was up-regulated in both the tail and intestine, two organs that undergo extensive transformations during metamorphosis and the focus of the current study. We then performed chromatin immunoprecipitation assays to investigate whether SRC3 is recruited to endogenous T3 target genes in vivo in developing tadpoles. Surprisingly, we found that SRC3 was recruited in a gene- and tissue-dependent manner to target genes by TR, both upon T3 treatment of premetamorphic tadpoles and during natural metamorphosis. In particular, in the tail, SRC3 was not recruited in a T3-dependent manner to the target TRbetaA promoter, suggesting either no recruitment or constitutive association. Finally, by using transgenic tadpoles expressing a dominant negative SRC3 (F-dnSRC3), we demonstrated that F-dnSRC3 was recruited in a T3-dependent manner in both the intestine and tail, blocking the recruitment of endogenous coactivators and histone acetylation. These results suggest that SRC3 is utilized in a gene- and tissue-specific manner by TR during development. PMID- 15901729 TI - A novel Porphyromonas gingivalis FeoB plays a role in manganese accumulation. AB - FeoB is an atypical transporter that has been shown to exclusively mediate ferrous ion transport in some bacteria. Unusually the genome of the periodontal pathogen Porphyromonas gingivalis has two genes (feoB1 and feoB2) encoding FeoB homologs, both of which are expressed in bicistronic operons. Kinetic analysis of ferrous ion transport by P. gingivalis W50 revealed the presence of a single, high affinity system with a K(t) of 0.31 microM. FeoB1 was found to be solely responsible for this transport as energized cells of the isogenic FeoB1 mutant (W50FB1) did not transport radiolabeled iron, while the isogenic FeoB2 mutant (W50FB2) transported radiolabeled iron at a rate similar to wild type. This was reflected in the iron content of W50FB1 grown in iron excess conditions which was approximately half that of the wild type and W50FB2. The W50FB1 mutant had increased sensitivity to both oxygen and hydrogen peroxide and was avirulent in an animal model of infection whereas W50FB2 exhibited the same virulence as the wild type. Analysis of manganous ion uptake using inductively coupled plasma-mass spectrometry revealed a greater than 3-fold decrease in intracellular manganese accumulation in W50FB2 which was also unable to grow in manganese-limited media. The protein co-expressed with FeoB2 appears to be a novel FeoA-MntR fusion protein that exhibits homology to a manganese-responsive, DNA-binding metalloregulatory protein. These results indicate that FeoB2 is not involved in iron transport but plays a novel role in manganese transport. PMID- 15901730 TI - Mammalian selenoprotein thioredoxin-glutathione reductase. Roles in disulfide bond formation and sperm maturation. AB - Thioredoxin reductases (TRs) are important redox regulatory enzymes, which control the redox state of thioredoxins. Mammals have cytosolic and mitochondrial TRs, which contain an essential selenocysteine residue and reduce cytosolic and mitochondrial thioredoxins. In addition, thioredoxin/glutathione reductase (TGR) was identified, which is a fusion of an N-terminal glutaredoxin domain and the TR module. Here we show that TGR is expressed at low levels in various tissues but accumulates in testes after puberty. The protein is particularly abundant in elongating spermatids at the site of mitochondrial sheath formation but is absent in mature sperm. We found that TGR can catalyze isomerization of protein and interprotein disulfide bonds and localized this function to its thiol domain. TGR targets include proteins that form structural components of the sperm, including glutathione peroxidase GPx4/PHGPx. Together, TGR and GPx4 can serve as a novel disulfide bond formation system. Both enzymes contain a catalytic selenocysteine consistent with the role of selenium in male reproduction. PMID- 15901731 TI - Identification of the VirB4-VirB8-VirB5-VirB2 pilus assembly sequence of type IV secretion systems. AB - Type IV secretion systems mediate the translocation of virulence factors (proteins and/or DNA) from Gram-negative bacteria into eukaryotic cells. A complex of 11 conserved proteins (VirB1-VirB11) spans the inner and the outer membrane and assembles extracellular T-pili in Agrobacterium tumefaciens. Here we report a sequence of protein interactions required for the formation of complexes between VirB2 and VirB5, which precedes their incorporation into pili. The NTPase Walker A active site of the inner membrane protein VirB4 is required for virulence, but an active site VirB4 variant stabilized VirB3 and VirB8 and enabled T-pilus formation. Analysis of VirB protein complexes extracted from the membranes with mild detergent revealed that VirB2-VirB5 complex formation depended on VirB4, which identified a novel T-pilus assembly step. Bicistron expression demonstrated direct interaction of VirB4 with VirB8, and analyses with purified proteins showed that VirB5 bound to VirB8 and VirB10. VirB4 therefore localizes at the basis of a trans-envelope interaction sequence, and by stabilization of VirB8 it mediates the incorporation of VirB5 and VirB2 into extracellular pili. PMID- 15901732 TI - The crucial role of trehalose and structurally related oligosaccharides in the biosynthesis and transfer of mycolic acids in Corynebacterineae. AB - Trehalose (alpha-D-glucopyranosyl-alpha'-D-glucopyranoside) is essential for the growth of the human pathogen Mycobacterium tuberculosis but not for the viability of the phylogenetically related corynebacteria. To determine the role of trehalose in the physiology of these bacteria, the so-called Corynebacterineae, mutant strains of Corynebacterium glutamicum unable to synthesize trehalose due to the knock-out of the genes of the three pathways of trehalose biosynthesis, were biochemically analyzed. We demonstrated that the synthesis of trehalose under standard conditions is a prerequisite for the production of mycolates, major and structurally important constituents of the cell envelope of Corynebacterineae. Consistently, the trehalose-less cells also lack the cell wall fracture plane that typifies mycolate-containing bacteria. Importantly, however, the mutants were able to synthesize mycolates when grown on glucose, maltose, and maltotriose but not on other carbon sources known to be used for the production of internal glucose phosphate such as fructose, acetate, and pyruvate. The mycoloyl residues synthesized by the mutants grown on alpha-D-glucopyranosyl containing oligosaccharides were transferred both onto the cell wall and free sugar acceptors. A combination of chemical analytical approaches showed that the newly synthesized glycolipids consisted of 1 mol of mycolate located on carbon 6 of the non reducing glucopyranosyl unit. Additionally, experiments with radioactively labeled trehalose showed that the transfer of mycoloyl residues onto sugars occurs outside the plasma membrane. Finally, and in contradiction to published data, we demonstrated that trehalose 6-phosphate has no impact on mycolate synthesis in vivo. PMID- 15901733 TI - Characterization of ChpBK, an mRNA interferase from Escherichia coli. AB - Escherichia coli contains a number of antitoxin-toxin modules on its chromosome, which are responsible for cell growth arrest and possible cell death. ChpBK is a toxin encoded by the ChpBIK antitoxin-toxin module. This module consists of a pair of genes, chpBI and chpBK encoding antitoxin ChpBI and toxin ChpBK, respectively. ChpBK consists of 116 amino acid residues, and its sequence shows 35% identity and 52% similarity to MazF, another E. coli toxin. MazF has been shown to be a sequence-specific (ACA) endoribonuclease that cleaves cellular mRNAs and effectively blocks protein synthesis and is thus termed as an mRNA interferase. Here we demonstrate that ChpBK is another mRNA interferase in E. coli whose induction effectively blocks cell growth in a manner similar to that of MazF. The protein synthesis as judged by incorporation of [35S]methionine was, however, reduced by only 60% upon ChpBK induction. We demonstrate that ChpBK is a new sequence-specific endoribonuclease that cleaves mRNAs both in vivo and in vitro at the 5'-or3'-side of the A residue in ACY sequences (Y is U, A, or G). The ChpBK cleavage of a synthetic RNA substrate generated a 2',3'-cyclic phosphate group at the 3'-end of the 5'-end product and a 5'-OH group at the 5' end of the 3'-end product in a manner identical to that of MazF. PMID- 15901734 TI - Y25S variant of Paracoccus pantotrophus cytochrome cd1 provides insight into anion binding by d1 heme and a rare example of a critical difference between solution and crystal structures. AB - Tyr25 is a ligand to the active site d1 heme in as isolated, oxidized cytochrome cd1 nitrite reductase from Paracoccus pantotrophus. This form of the enzyme requires reductive activation, a process that involves not only displacement of Tyr25 from the d1 heme but also switching of the ligands at the c heme from bis histidinyl to His/Met. A Y25S variant retains this bis-histidinyl coordination in the crystal of the oxidized state that has sulfate bound to the d1 heme iron. This Y25S form of the enzyme does not require reductive activation, an observation previously interpreted as meaning that the presence of the phenolate oxygen of Tyr25 is the critical determinant of the requirement for activation. This interpretation now needs re-evaluation because, unexpectedly, the oxidized as prepared Y25S protein, unlike the wild type, has different heme iron ligands in solution at room temperature, as judged by magnetic circular dichroism and electron spin resonance spectroscopies, than in the crystal. In addition, the binding of nitrite and cyanide to oxidized Y25S cytochrome cd1 is markedly different from the wild type enzyme, thus providing insight into the affinity of the oxidized d1 heme ring for anions in the absence of the steric barrier presented by Tyr25. PMID- 15901735 TI - Macrophages survive hyperoxia via prolonged ERK activation due to phosphatase down-regulation. AB - Macrophages exposed to hyperoxia in the lung continue to survive for prolonged periods. We previously reported (Nyunoya, T., Powers, L. S., Yarovinsky, T. O., Butler, N. S., Monick, M. M., and Hunninghake, G. W. (2003) J. Biol. Chem. 278, 36099-36106) that hyperoxia induces cell cycle arrest and sustained extracellular signal-related kinase (ERK) activity in macrophages. In this study, we determined the mechanisms of hyperoxia-induced ERK activation and how ERK activity plays a pro-survival role in hyperoxia-exposed cells. Inhibition of ERK activity decreased survival of hyperoxia-exposed macrophages. This was due, at least in part, to down-regulation of the pro-apoptotic Bcl-2 family member, BimEL. In determining the mechanism of ERK activation by hyperoxia, we found that ERK activation was not associated with hyperoxia-induced activation of the upstream ERK kinase mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1/2. When we examined the ability of whole cell lysates from hyperoxia exposed cells to dephosphorylate purified phosphorylated ERK, we found decreased ERK-directed phosphatase activity. Two particular ERK-directed phosphatases (protein phosphatase 2A and MAPK phosphatase-3) demonstrated decreased activity in hyperoxia-exposed cells. Moreover, whole cell lysates from normoxia-exposed cells depleted of PP2A or MAPK phosphatase-3 were also less able to dephosphorylate ERK. These data demonstrate that, in hyperoxia-exposed macrophages, sustained activation of ERK due to phosphatase down-regulation permits macrophage survival via effects on the balance between pro- and anti apoptotic Bcl-2 family proteins. PMID- 15901736 TI - Inefficient maturation of the rat luteinizing hormone receptor. A putative way to regulate receptor numbers at the cell surface. AB - Increasing evidence suggests that the folding and maturation of monomeric proteins and assembly of multimeric protein complexes in the endoplasmic reticulum (ER) may be inefficient not only for mutants that carry changes in the primary structure but also for wild type proteins. In the present study, we demonstrate that the rat luteinizing hormone receptor, a G protein-coupled receptor, is one of these proteins that matures inefficiently and appears to be very prone to premature degradation. A substantial portion of the receptors in stably transfected human embryonic kidney 293 cells existed in immature form of M(r) 73,000, containing high mannose-type N-linked glycans. In metabolic pulse chase studies, only approximately 20% of these receptor precursors were found to gain hormone binding ability and matured to a form of M(r) 90,000, containing bi- and multiantennary sialylated N-linked glycans. The rest had a propensity to form disulfide-bonded complexes with a M(r) 120,000 protein in the ER membrane and were eventually targeted for degradation in proteasomes. The number of membrane bound receptor precursors increased when proteasomal degradation was inhibited, and no cytosolic receptor forms were detected, suggesting that retrotranslocation of the misfolded/incompletely folded receptors is tightly coupled to proteasomal function. Furthermore, a proteasomal blockade was found to increase the number of receptors that were capable of hormone binding. Thus, these results raise the interesting possibility that luteinizing hormone receptor expression at the cell surface may be controlled at the ER level by regulating the number of newly synthesized proteins that will mature and escape the ER quality control and premature degradation. PMID- 15901737 TI - Three distinct molecular surfaces in ephrin-A5 are essential for a functional interaction with EphA3. AB - Eph receptor tyrosine kinases (Ephs) function as molecular relays that interact with cell surface-bound ephrin ligands to direct the position of migrating cells. Structural studies revealed that, through two distinct contact surfaces on opposite sites of each protein, Eph and ephrin binding domains assemble into symmetric, circular heterotetramers. However, Eph signal initiation requires the assembly of higher order oligomers, suggesting additional points of contact. By screening a random library of EphA3 binding-compromised ephrin-A5 mutants, we have now determined ephrin-A5 residues that are essential for the assembly of high affinity EphA3 signaling complexes. In addition to the two interfaces predicted from the crystal structure of the homologous EphB2.ephrin-B2 complex, we identified a cluster of 10 residues on the ephrin-A5 E alpha-helix, the E-F loop, the underlying H beta-strand, as well as the nearby B-C loop, which define a distinct third surface required for oligomerization and activation of EphA3 signaling. Together with a corresponding third surface region identified recently outside of the minimal ephrin binding domain of EphA3, our findings provide experimental evidence for the essential contribution of three distinct protein interaction interfaces to assemble functional EphA3 signaling complexes. PMID- 15901738 TI - Direct binding to ceramide activates protein kinase Czeta before the formation of a pro-apoptotic complex with PAR-4 in differentiating stem cells. AB - We have reported that ceramide mediates binding of atypical protein kinase C (PKC) zeta to its inhibitor protein, PAR-4 (prostate apoptosis response-4), thereby inducing apoptosis in differentiating embryonic stem cells. Using a novel method of lipid vesicle-mediated affinity chromatography, we showed here that endogenous ceramide binds directly to the PKCzeta.PAR-4 complex. Ceramide and its analogs activated PKCzeta prior to binding to PAR-4, as determined by increased levels of phosphorylated PKCzeta and glycogen synthase kinase-3beta and emergence of a PAR-4-to-phosphorylated PKCzeta fluorescence resonance energy transfer signal that co-localizes with ceramide. Elevated expression and activation of PKCzeta increased cell survival, whereas expression of PAR-4 promoted apoptosis. This suggests that PKCzeta counteracts apoptosis, unless its ceramide-induced activation is compromised by binding to PAR-4. A luciferase reporter assay showed that ceramide analogs activate nuclear factor (NF)-kappaB unless PAR-4-dependent inhibition of PKCzeta suppresses NF-kappaB activation. Taken together, our results show that direct physical association with ceramide and PAR-4 regulates the activity of PKCzeta. They also indicate that this interaction regulates the activity of glycogen synthase kinase-3beta and NF-kappaB. PMID- 15901739 TI - Point mutational analysis of the liganding site in human glycolipid transfer protein. Functionality of the complex. AB - Mammalian glycolipid transfer proteins (GLTPs) facilitate the selective transfer of glycolipids between lipid vesicles in vitro. Recent structural determinations of the apo- and glycolipid-liganded forms of human GLTP have provided the first insights into the molecular architecture of the protein and its glycolipid binding site (Malinina, L., Malakhova, M. L., Brown, R. E., and Patel, D. J. (2004) Nature 430, 1048-1053). In the present study, we have evaluated the functional consequences of point mutation of the glycolipid liganding site of human GLTP within the context of a carrier-based mechanism of glycolipid intermembrane transfer. Different approaches were developed to rapidly and efficiently assess the uptake and release of glycolipid by GLTP. They included the use of glass-immobilized, glycolipid films to load GLTP with glycolipid and separation of GLTP/glycolipid complexes from vesicles containing glycolipid (galactosylceramide or lactosylceramide) or from monosialoganglioside dispersions by employing nickel-nitrilotriacetic acid-based affinity or gel filtration strategies. Point mutants of the sugar headgroup recognition center (Trp-96, Asp 48, Asn-52) and of the ceramide-accommodating hydrophobic tunnel (Phe-148, Phe 183, Leu-136) were analyzed for their ability to acquire and release glycolipid ligand. Two manifestations of point mutation within the liganding site were apparent: (i) impaired formation of the GLTP/glycolipid complex; (ii) impaired acquisition and release of bound glycolipid by GLTP. The results are consistent with a carrier-based mode of GLTP action to accomplish the intermembrane transfer of glycolipid. Also noteworthy was the inefficient release of glycolipid by wtGLTP into phosphatidylcholine acceptor vesicles, raising the possibility of a function other than intermembrane glycolipid transfer in vivo. PMID- 15901740 TI - Mutational and structural analyses of the hinge region of membrane type 1-matrix metalloproteinase and enzyme processing. AB - Membrane type 1 (MT1)-matrix metalloproteinase (MMP) is a major mediator of collagen degradation in the pericellular space in both physiological and pathological conditions. Previous evidence has shown that on the cell surface, active MT1-MMP undergoes autocatalytic processing to a major membrane-tethered 44 kDa product lacking the catalytic domain and displaying Gly285 at its N terminus, which is at the beginning of the hinge domain. However, the importance of this site and the hinge region in MT1-MMP processing is unknown. In the current study, we generated mutations and deletions in the hinge of MT1-MMP and followed their effect on processing. These studies established Gly284-Gly285 as the main cleavage site involved in the formation of the 44-kDa species. However, alterations at this site did not prevent processing. Instead, they forced downstream cleavages within the stretch of residues flanked by Gln296 and Ser304 in the hinge region, as determined by the processing profile of various hinge deletion mutants. Also, replacement of the hinge of MT1-MMP with the longer MT3 MMP hinge did not prevent processing of MT1-MMP. Molecular dynamic studies using a computational model of MT1-MMP revealed that the hinge region is a highly motile element that undergoes significant motion in the highly exposed loop formed by Pro295-Arg302 consistent with being a prime target for proteolysis, in agreement with the mutational data. These studies suggest that the hinge of MT1 MMP evolved to facilitate processing, a promiscuous but compulsory event in the destiny of MT1-MMP, which may play a key role in the control of pericellular proteolysis. PMID- 15901741 TI - BAD detects coincidence of G2/M phase and growth factor deprivation to regulate apoptosis. AB - BAD, a member of the Bcl-2 protein family, promotes mitochondria-dependent apoptosis. Here, we report that BAD dissociates from 14-3-3zeta at each G2/M phase of proliferating lymphoid cells. The cell cycle-dependent dissociation of BAD was associated with phosphorylation at Ser-128, whereas mutant S128A-BAD, in which Ser-128 was converted to alanine, remained associated with 14-3-3zeta throughout the cell cycle. Although the cell cycle-dependent dissociation of BAD per se did not induce apoptosis, growth factor deprivation induced prompt apoptosis at the G2/M phase but not at the G1 phase. In cells expressing S128A BAD, growth factor deprivation-induced apoptosis was markedly delayed and was accompanied by a delayed dephosphorylation of growth factor-dependent regulatory serine residues. These results indicate that BAD induces apoptosis upon detecting the coincidence of G2/M phase and growth factor deprivation. PMID- 15901742 TI - Over-the-counter progesterone cream produces significant drug exposure compared to a food and drug administration-approved oral progesterone product. AB - Progesterone products are available in prescription form as well as over-the counter (OTC) topical preparations sold for "cosmetic" uses. In a randomized study design, the authors compared the drug exposure from an OTC progesterone cream to a Food and Drug Administration-approved oral preparation at the labeled daily doses recommended for each product. Twelve healthy postmenopausal women received 200-mg oral progesterone capsules once daily for 12 days or progesterone cream 40 mg twice daily for 12 days. At steady state (day 12 of each phase), whole-blood samples were collected over 24 hours (oral progesterone) or 12 hours (topical progesterone) and assayed for total progesterone concentration. No significant differences were found in dose-normalized 24-hour progesterone exposure comparing the cream to oral capsules (median AUC(0-24) 12.5 ng x h/mL vs 10.5 ng x h/mL, respectively; P = .81). In light of the potential risks associated with long-term progesterone use, the authors question whether topical progesterone products should be available OTC. PMID- 15901743 TI - Pharmacokinetics, safety, and tolerability of intramuscular ziprasidone in healthy volunteers. AB - Little has been published regarding the pharmacokinetics of the intramuscular (IM) formulation of Ziprasidone. The authors report results from 2 early phase I studies in healthy volunteers: a trial of single 5-, 10-, or 20-mg IM doses of ziprasidone in 24 subjects and an open-label 3-way crossover trial of 5-mg intravenous (IV), 5-mg IM, and 20-mg oral ziprasidone in 12 subjects. Absorption of IM ziprasidone was rapid (Tmax < 1 hour). The IM pharmacokinetic profile was consistent between studies and linear, with dose-related increases in exposure observed. The mean IM elimination t(1/2) was short and approximately 2.5 hours. The mean bioavailability for the 5-mg IM ziprasidone dose was approximately 100%. Adverse events were generally mild to moderate, and no subjects were discontinued from the study. No significant effects on renal function or other laboratory values were noted. These results support the use of IM ziprasidone in treating acutely agitated patients with schizophrenia. PMID- 15901744 TI - Population pharmacokinetics of eniporide and its metabolite in healthy subjects and patients with acute myocardial infarction. AB - Eniporide (EMD 96 875) is a novel and selective inhibitor of the Na+-H+ exchange (NHE-1) inhibitor. The study objectives were to identify a structural model for population pharmacokinetic analysis of eniporide and its metabolite (EMD 112 843) using nonlinear mixed-effects modeling after short-term infusion (dose: 2.5-400 mg) in healthy subjects and patients undergoing myocardial reperfusion therapy. Pooled concentrations of eniporide and its metabolite from healthy subjects (n = 153; 4815 observations) and patients (n = 304; 1465 observations) were included in the pharmacokinetic analysis. Population estimates of clearance and volume of distribution of eniporide were 29.2 L/h (24.1% coefficient of variation [CV], healthy), 20.8 L/h (28.0% CV, patients) and 20.4 L (13.1% CV, healthy), 16.9 L (24.9% CV, patients), respectively. Statistical significance was achieved for the effect of age on clearance and creatinine clearance on volume of distribution of eniporide. The impact of the covariates on eniporide pharmacokinetics is minimal to warrant any dosage adjustments in patient population. PMID- 15901745 TI - Steady-state serum concentrations of progesterone following continuous intravenous infusion in patients with acute moderate to severe traumatic brain injury. AB - Progesterone (PG) has been shown to provide substantial neuroprotection after traumatic brain injury (TBI) in multiple animal models. As a first step in assessing applicability to humans, the authors examined the effects of acute TBI and extracranial trauma on the pharmacokinetics of PG given by intravenous infusion. Multiple blood samples were obtained from 11 female and 21 male trauma patients receiving PG and 1 female and 3 male patients receiving placebo infusions for 72 hours. Values for C(SS), CL, t(1/2), and Vd were obtained using AUC((0-72)) and postinfusion blood samples. C(SS) values were 337 +/- 135 ng/mL, which were significantly lower than the target concentration of 450 +/- 100 ng/mL. The lower C(SS) is attributed to the CL, which was higher than anticipated. In addition, t(1/2) was longer and V(d) was higher than anticipated. These results demonstrate that stable PG concentrations can be rapidly achieved following TBI. PMID- 15901746 TI - A single administration of recombinant human interleukin-12 is associated with increased expression levels of interferon-gamma and signal transducer and activator of transcription in healthy subjects. AB - The objectives of this study were to assess the safety and tolerability of single doses of 1, 4, and 8 mug of recombinant human interleukin-12 (rhIL-12) administered subcutaneously to healthy subjects. The pharmacokinetics, pharmacodynamics, and pharmacogenomics of rhIL-12 were evaluated. Recombinant human IL-12 was well tolerated in these healthy male and female subjects. The most frequently reported adverse events were flu-like symptoms, which exhibited a dose-response relationship. Pharmacokinetic analysis suggested that serum IL-12 levels increased with dose. Analysis of serum levels indicated that interferon gamma increased with the dose of rhIL-12, whereas IL-6 levels showed no changes with rhIL-12 treatment. The messenger ribonucleic acid expression of signal transducer and activator of transcription was significantly increased 24 hours after the administration of rhIL-12 for all dose groups versus placebo, and results indicated that the magnitude of increase may be dose dependent. This study suggests that interferon-gamma and signal transducer and activator of transcription are biomarkers of rhIL-12 activity. PMID- 15901747 TI - Penetration of ertapenem into different pulmonary compartments of patients undergoing lung surgery. AB - Ertapenem is approved for the treatment of community-acquired pneumonia (CAP), but its in vivo penetration into lung tissue (LT), epithelial lining fluid (ELF), and alveolar cells (AC) is unknown. Fifteen patients undergoing thoracotomy were treated with 1 g intravenously for perioperative prophylaxis. Bronchoalveolar lavage was performed 1, 3, and 5 hours after ertapenem infusion. Normal LT was sampled at the time of lung extraction. Blood was collected before and at different time points up to 24 hours after infusion. Mean concentrations of ertapenem in plasma, ELF, and AC were at 1.0 hour, 63.1, 4.06, 0.004 mg/L; at 3.0 hours, 39.7, 2.59, 0.003 mg/L; and at 5.0 hours, 27.2, 2.83, 0.007 mg/L. Mean (range) concentration in LT was 7.60 (2.5-19.4) mg/kg tissue 1.5 to 4.5 hours after infusion. In plasma, ertapenem exhibited a Cmax of 94.7 +/- 23.3 mg/L and an AUC(0-last) of 501.1 +/- 266.3 mg x h/L. These results, combined with the reported (MIC)90 of most CAP bacteria, support the previously observed clinical efficacy of ertapenem in the treatment of CAP. PMID- 15901748 TI - Effect of age and postoperative time on cytochrome p450 enzyme activity following liver transplantation. AB - This study evaluates the changes in cytochrome P450 (CYP) enzyme activity in orthotopic liver transplant (OLTx) patients in relation to recipient age and postoperative time. Thirty-eight stable OLTx patients, separated into younger and older age groups, and 21 healthy subjects were given a 5-drug cocktail including chlorzoxazone (CYP2E1), caffeine (CYP1A2), dapsone (CYP3A4), mephenytoin (CYP2C19), and debrisoquin (CYP2D6). The phenotypic indexes were determined for each associated enzyme. Compared to young healthy subjects, the CYP2E1 capacity was significantly increased in younger and older OLTx patients (P < .001), while the CYP2C19 capacity was decreased significantly in younger and older OLTx patients within 30 days postoperatively (P < .01). The CYP2D6 capacity was significantly lower after 30 days postoperatively in older OLTx patients (P < .05). The authors conclude that within 30 days postoperatively, CYP2E1 capacity was markedly elevated in OLTx patients, while 2C19 function was significantly reduced. CYP2D6 capacity was impaired after 30 days postoperatively. Younger and older OLTx patients experienced similar changes in major CYP450 enzyme capacity following liver transplantation. PMID- 15901749 TI - Pharmacokinetics of paclitaxel in ovarian cancer patients and genetic polymorphisms of CYP2C8, CYP3A4, and MDR1. AB - Interindividual differences in the pharmacokinetics of paclitaxel and its metabolites in Japanese ovarian cancer patients were investigated in relation to genetic polymorphisms of the CYP2C8, CYP3A4, and MDR1 genes. The area under the concentration-time curve (AUC) ratios of paclitaxel/6alpha-hydroxypaclitaxel and paclitaxel/3 -p-hydroxypaclitaxel calculated as the metabolic index of CYP2C8 and CYP3A4 showed 13- and 12-fold interindividual variations, respectively. No patient had any CYP2C8 variants, while 2 patients were heterozygotes of CYP3A4*16. For the MDR1 gene, the frequencies of -129C, 1236C, 2677T, 2677A, and 3435T alleles were 2.2%, 8.7%, 56.5%, 4.4%, and 52.2%, respectively. Subjects possessing the 3435T allele had a significantly (P < .05) higher AUC of 3'- p hydroxypaclitaxel compared to those possessing the 3435C allele. Leukocytopenia was significantly (P < .05) related to the AUC of paclitaxel. Genotyping of the CYP2C8, CYP3A4, and MDR1 genes might not be essential to predict adverse effects of paclitaxel in Japanese patients, although an allelic variant of MDR1 may functionally affect the pharmacokinetics of its metabolite. PMID- 15901750 TI - Bioavailability and pharmacokinetics of alkamides from the roots of Echinacea angustifolia in humans. AB - Alkamides are suspected to contribute to the activity of Echinacea preparations. They are mainly derived from undeca- and dodecanoic acid and differ in the degree of unsaturation and the configuration of the double bonds. In total, 6 alkamides have been isolated from the roots of Echinacea angustifolia as major lipophilic constituents and have been investigated regarding their pharmacokinetics. A sensitive and specific method has been developed for the identification and quantification of these alkamides in human plasma using liquid chromatography electrospray ionization ion-trap mass spectrometry. The method was applied to analyze plasma samples obtained from a randomized, open, single-dose, crossover study after oral administration of a 60% ethanolic extract from the roots of E. angustifolia to 11 healthy subjects. The maximum concentration of dodeca 2E,4E,8Z,10E/Z-tetraenoic acid isobutylamides, the main alkamides in the roots of E. angustifolia, appeared already after 30 minutes and was 10.88 ng/mL for the 2.5-mL dose. PMID- 15901751 TI - Lack of significant effect of grapefruit juice on the pharmacokinetics of lansoprazole and its metabolites in subjects with different CYP2C19 genotypes. PMID- 15901752 TI - Bioavailability of amlodipine besylate following oral administration as a tablet dispersed in applesauce. PMID- 15901753 TI - The effect of ketoconazole on the pharmacokinetics of a selective alpha 1A adrenoceptor antagonist. PMID- 15901754 TI - Utility of levosimendan, a new calcium sensitizing agent, in the treatment of cardiogenic shock due to myocardial stunning in patients with ST-elevation myocardial infarction: a series of cases. PMID- 15901755 TI - Interaural translational VOR: suppression, enhancement, and cognitive control. AB - We investigated the influence of cognitive factors on the early response of the interaural translational vestibuloocular reflex (tVOR) in six normal subjects. Variables were prior knowledge of direction of head motion and the position of the fixation target relative to the head [head-fixed (HF) or space-fixed (SF)]. A manually driven device provided a step-like head translation (approximately 35 mm distance, peak acceleration, 0.6-1.3 g). Subjects looked at the SF or HF target located 15 cm in front of their heads in otherwise complete darkness. The testing paradigms were: random interleaving of SF and HF targets with unknown direction of head movement, known target location with random head direction (SFR or HFR), and known target location with known head direction (SFP or HFP). Timing was always unpredictable. A "gain" of the slow phase was calculated with respect to ideal performance (maintained fixation of the SF target, recorded/ideal eye velocity computed at time of peak head velocity). At such times, there were no significant differences in gain between HF and SF trials in the random condition; the average gain was approximately 36% of ideal. On the other hand, responses in the SFR and HFR conditions differed as early as 20 ms after the head began moving. Average gain was higher (0.43 +/- 0.11 vs. 0.34 +/- 0.14; means +/- SD, P < 0.05) for each subject in the SFR than the HFR condition. For SFP and HFP, the responses differed from the onset of head motion. Average slow-phase gain was higher (0.49 +/- 0.12 vs. 0.31 +/- 0.12, P < 0.02) for each subject in SFP than in HFP. The timing of corrective saccades during the tVOR was also influenced by cognitive factors. Visual error signals seemed to be more important for triggering saccades in HF trials, whereas preprogramming, probably based on labyrinthine information, seemed to be more important in SF trials. Simulations showed that the changes in slow-phase gain with cognition could be reproduced with simple parametric adjustments of the gain of activity from otolith afferents and suggest that higher-level cognitive control of the VOR could occur as early as the synapse of peripheral afferents on neurons in the vestibular nuclei, either directly from higher level centers or via the cerebellum. In sum, the tVOR both in its slow-phase response and the saccadic corrections-is subject to "higher-level" cognitive influences including knowledge of where the line of sight must point during head motion and the impending direction of head motion. PMID- 15901756 TI - Sex differences in the cannabinoid modulation of an A-type K+ current in neurons of the mammalian hypothalamus. AB - Cannabinoids regulate biological processes governed by the hypothalamus including, but not limited to, energy homeostasis and reproduction. The present study sought to determine whether cannabinoids modulate A-type K(+) currents (I(A)) in neurons of the hypothalamic arcuate nucleus (ARC). Whole cell patch clamp recordings were performed in slices through the ARC prepared from castrated female and male guinea pigs. Forty percent of guinea pig ARC neurons exhibited a transient outward current that was antagonized by high (mM) concentrations of 4 aminopyridine and (100 nM) rHeteropodatoxin-2. Five of these neurons also were immunopositive for both beta-endorphin and the Kv4.2 channel subunit. Bath application of the CB1 receptor agonists WIN 55,212-2 (1 microM) or ACEA (1 microM) selectively induced a rightward shift in the inactivation curve for the I(A), significantly increasing the half-maximal voltage without affecting the peak current magnitude, in neurons from female but not male animals. The CB1 receptor antagonist AM251 (1 microM) reversed this action. Collectively, these data reveal that guinea pig ARC neurons, including proopiomelanocortin neurons, express a prominent I(A) that is positively modulated by cannabinoids in a sex specific way by altering the voltage dependence of its inactivation. The resultant inhibitory effect on this neuronal population may shed some insight into the mechanism(s) by which cannabinoids influence hypothalamic function. PMID- 15901757 TI - Synaptic transmission at the cochlear nucleus endbulb synapse during age-related hearing loss in mice. AB - Age-related hearing loss (AHL) typically starts from high-frequency regions of the cochlea and over time invades lower-frequency regions. During this progressive hearing loss, sound-evoked activity in spiral ganglion cells is reduced. DBA mice have an early onset of AHL. In this study, we examined synaptic transmission at the endbulb of Held synapse between auditory nerve fibers and bushy cells in the anterior ventral cochlear nucleus (AVCN). Synaptic transmission in hearing-impaired high-frequency areas of the AVCN was altered in old DBA mice. The spontaneous miniature excitatory postsynaptic current (mEPSC) frequency was substantially reduced (about 60%), and mEPSCs were significantly slower (about 115%) and smaller (about 70%) in high-frequency regions of old (average age 45 days) DBA mice compared with tonotopically matched regions of young (average age 22 days) DBA mice. Moreover, synaptic release probability was about 30% higher in high-frequency regions of young DBA than that in old DBA mice. Auditory nerve-evoked EPSCs showed less rectification in old DBA mice, suggesting recruitment of GluR2 subunits into the AMPA receptor complex. No similar age-related changes in synaptic release or EPSCs were found in age matched, normal hearing young and old CBA mice. Taken together, our results suggest that auditory nerve activity plays a critical role in maintaining normal synaptic function at the endbulb of Held synapse after the onset of hearing. Auditory nerve activity regulates both presynaptic (release probability) and postsynaptic (receptor composition and kinetics) function at the endbulb synapse after the onset of hearing. PMID- 15901758 TI - Pain and temperature encoding in the human thalamic somatic sensory nucleus (Ventral caudal): inhibition-related bursting evoked by somatic stimuli. AB - Stimulus-evoked inhibitory events have not been demonstrated in thalamic spike trains encoding of pain and temperature stimuli. We have now tested the hypothesis that the human thalamic response to mechanical and thermal stimuli is characterized by low-threshold calcium spike (LTS)-associated bursts of high frequency action potentials preceded by prolonged inhibition. The results included 57 neurons recorded in the human thalamic principal somatic sensory nucleus (ventral caudal, Vc) of 24 patients during awake surgery. Neurons were classified by the grading of their response with stimulus intensity into the painful range (graded or non-graded) and the stimulus response (to mechanical, cold, or heat stimuli). Firing rates were analyzed by the response to all stimuli combined (stimuli overall) and to the stimulus characteristic of the stimulus response type (optimal stimulus), e.g., cold stimuli for neurons of the cold stimulus response type. All neuronal categories had clear stimulus-evoked LTS bursting as identified by the criteria for selecting bursts in the spike train, by significant preburst inhibition, and by preburst inter-spike interval not significantly <100 ms. Stimulus-evoked LTS burst rates were significantly higher for neurons in the cold stimulus response type independent of the firing rate between bursts. The parameters of preburst inhibition were largely independent of the neuronal category and the stimuli included in the analysis, which suggests inhibitory mechanisms are similar across neuronal types. Therefore LTS bursting is a substantial, nonlinear component of the spontaneous and stimulus-evoked activity of thalamic neurons in awake humans. PMID- 15901759 TI - Population coding of self-motion: applying bayesian analysis to a population of visual interneurons in the fly. AB - Coding of sensory information often involves the activity of neuronal populations. We demonstrate how the accuracy of a population code depends on integration time, the size of the population, and noise correlation between the participating neurons. The population we study consists of 10 identified visual interneurons in the blowfly Calliphora vicina involved in optic flow processing. These neurons are assumed to encode the animal's head or body rotations around horizontal axes by means of graded potential changes. From electrophysiological experiments we obtain parameters for modeling the neurons' responses. From applying a Bayesian analysis on the modeled population response we draw three major conclusions. First, integration of neuronal activities over a time period of only 5 ms after response onset is sufficient to decode accurately the rotation axis. Second, noise correlation between neurons has only little impact on the population's performance. And third, although a population of only two neurons would be sufficient to encode any horizontal rotation axis, the population of 10 vertical system neurons is advantageous if the available integration time is short. For the fly, short integration times to decode neuronal responses are important when controlling rapid flight maneuvers. PMID- 15901760 TI - An oscillatory hierarchy controlling neuronal excitability and stimulus processing in the auditory cortex. AB - EEG oscillations are hypothesized to reflect cyclical variations in the neuronal excitability, with particular frequency bands reflecting differing spatial scales of brain operation. However, despite decades of clinical and scientific investigation, there is no unifying theory of EEG organization, and the role of ongoing activity in sensory processing remains controversial. This study analyzed laminar profiles of synaptic activity [current source density CSD] and multiunit activity (MUA), both spontaneous and stimulus-driven, in primary auditory cortex of awake macaque monkeys. Our results reveal that the EEG is hierarchically organized; delta (1-4 Hz) phase modulates theta (4-10 Hz) amplitude, and theta phase modulates gamma (30-50 Hz) amplitude. This oscillatory hierarchy controls baseline excitability and thus stimulus-related responses in a neuronal ensemble. We propose that the hierarchical organization of ambient oscillatory activity allows auditory cortex to structure its temporal activity pattern so as to optimize the processing of rhythmic inputs. PMID- 15901761 TI - Endogenous ATP involvement in mustard-oil-induced central sensitization in trigeminal subnucleus caudalis (medullary dorsal horn). AB - Central sensitization represents a sustained hypersensitive state of dorsal horn nociceptive neurons that can be evoked by peripheral inflammation or injury to nerves and tissues. It reflects neuroplastic changes such as increases in neuronal spontaneous activity, receptive field size, and responses to suprathreshold stimuli and a decrease in activation threshold. We recently demonstrated that purinergic receptor mechanisms in trigeminal subnucleus caudalis (Vc; medullary dorsal horn) are also involved in the initiation and maintenance of central sensitization in brain stem nociceptive neurons of trigeminal subnucleus oralis. The aim of the present study was to investigate whether endogenous ATP is involved in the development of central sensitization in Vc itself. The experiments were carried out on urethan/alpha-chloralose anesthetized and immobilized rats. Single neurons were recorded and identified as nociceptive-specific (NS) in the deep laminae of Vc. During continuous saline superfusion (0.6 ml/h it) over the caudal medulla, Vc neuronal central sensitization was readily induced by mustard oil application to the tooth pulp. However, this mustard-oil-induced central sensitization could be completely blocked by continuous intrathecal superfusion of the wide-spectrum P2X receptor antagonist pyridoxal-phosphate-6-azophenyl-2, 4-disulphonic acid tetra-sodium (33 100 microM) and by apyrase (an ectonucleotidase enzyme, 30 units/ml). Superfusion of the selective P2X1, P2X3 and P2X(2/3) receptor antagonist 2',3'-O-(2,4,6 trinitrophenyl) adenosine 5'-triphosphate (6-638 microM) partially blocked the Vc central sensitization. The two P2X receptor antagonists did not significantly affect the baseline nociceptive properties of the Vc neurons. These findings implicate endogenous ATP as an important mediator contributing to the development of central sensitization in nociceptive neurons of the deep laminae of the dorsal horn. PMID- 15901762 TI - BDNF-induced facilitation of afferent-evoked responses in lamina II neurons is reduced after neonatal spinal cord contusion injury. AB - We previously reported that brain-derived neurotrophic factor (BDNF), a pronociceptive neurotransmitter, induces synaptic facilitation of excitatory postsynaptic current (EPSC) in lamina II neurons of neonatal rats up to P14 in a N-methyl-d-aspartate (NMDA) receptor-dependent manner. Here we used the patch clamp technique to study synaptic and NMDA-evoked responses in transverse spinal slices in the lumbar enlargement as well as the ability of BDNF to modify these responses from 1 day to 6 wk after neonatal contusion. In older uninjured animals (>P14), BDNF continued to evoke synaptic facilitation although superfusion of NMDA (in TTX) induced inward current of significantly smaller amplitude than that observed in younger rats. After contusion injury, BDNF was unable to facilitate dorsal root-evoked EPSCs in lamina II neurons despite the finding that NMDA evoked currents were only slightly smaller than those observed in age-matched uninjured animals. These findings suggest that although BDNF-induced facilitation of the AMPA/kainate receptor-mediated response to dorsal root stimulation is maintained in the mature dorsal horn from intact rats, BDNF may no longer elicit these pronociceptive actions after neonatal contusion injury. The lack of change in NMDA-evoked currents in contused cords suggests that diminished NMDA receptor function is not the major cause of the decline in BDNF action after contusion. It seems more likely that diminished trkB expression and enhanced expression of truncated trkB receptors in the contused cord play a significant role in determining the reduced effect of BDNF under these conditions. PMID- 15901763 TI - Sam68-like mammalian protein 2, identified by digital differential display as expressed by podocytes, is induced in proteinuria and involved in splice site selection of vascular endothelial growth factor. AB - Podocytes, the glomerular epithelial cells of the kidney, share important features with neuronal cells. In addition to phenotypical and functional similarities, a number of gene products have been found to be expressed exclusively or predominantly by both cell types. With the hypothesis of a common transcriptome shared by podocytes and neurons, digital differential display was used to identify novel podocyte-expressed gene products. Comparison of brain and kidney cDNA libraries with those of other organs identified Sam68-like mammalian protein 2 (SLM-2), a member of the STAR family of RNA processing proteins, as expressed by podocytes. SLM-2 expression was found to be restricted in the kidney to podocytes. In proteinuric diseases, SLM-2, a known regulator of neuronal mRNA splice site selection, was found significantly upregulated on mRNA and protein levels. Knockdown of SLM-2 by short interfering RNA in podocytes was performed to evaluate its biologic role. RNA splicing of vascular endothelial growth factor (VEGF), a key regulator of the filtration barrier and expressed as functionally distinct splice isoforms, was evaluated. VEGF(165) expression was found to be reduced by 25% after SLM-2 knockdown. In vivo, the glomerular expression of SLM-2 correlated with the mRNA levels of VEGF(165). This study demonstrates the power of digital differential display to predict cell type-specific gene expression by hypothesis-driven analysis of tissue cDNA libraries. SLM-2-dependent VEGF splicing indicates the importance of mRNA splice site selection for glomerular filtration barrier function. PMID- 15901764 TI - Metabolic syndrome and the risk for chronic kidney disease among nondiabetic adults. AB - The metabolic syndrome is a risk factor for the development of diabetes and cardiovascular disease; however, no prospective studies have examined the metabolic syndrome as a risk factor for chronic kidney disease (CKD). A total of 10,096 nondiabetic participants who were in the Atherosclerosis Risk in Communities study and had normal baseline kidney function composed the study cohort. The metabolic syndrome was defined according to recent guidelines from the National Cholesterol Education Program. Incident CKD was defined as an estimated GFR (eGFR) <60 ml/min per 1.73 m2 at study year 9 among those with an eGFR > or =60 ml/min per 1.73 m2 at baseline. After 9 yr of follow-up, 691 (7%) participants developed CKD. The multivariable adjusted odds ratio (OR) of developing CKD in participants with the metabolic syndrome was 1.43 (95% confidence interval [CI], 1.18 to 1.73). Compared with participants with no traits of the metabolic syndrome, those with one, two, three, four, or five traits of the metabolic syndrome had OR of CKD of 1.13 (95% CI, 0.89 to 1.45), 1.53 (95% CI, 1.18 to 1.98), 1.75 (95% CI, 1.32 to 2.33), 1.84 (95% CI, 1.27 to 2.67), and 2.45 (95% CI, 1.32 to 4.54), respectively. After adjusting for the subsequent development of diabetes and hypertension during the 9 yr of follow-up, the OR of incident CKD among participants with the metabolic syndrome was 1.24 (95% CI, 1.01 to 1.51). The metabolic syndrome is independently associated with an increased risk for incident CKD in nondiabetic adults. PMID- 15901765 TI - Protection against renal ischemia reperfusion injury by CD44 disruption. AB - Inflammation contributes to renal ischemia reperfusion (I/R) injury, potentially causing renal dysfunction. The inflammatory infiltrate mainly consists of neutrophils, which are deleterious for the renal tissue. Because CD44 is expressed by neutrophils and is rapidly upregulated by capillary endothelial cells after I/R injury, it was hypothesized that CD44 might play an important role in the development of I/R injury. This study showed that rapid CD44 upregulation on renal capillary endothelial cells mediates neutrophil recruitment to the postischemic tissue. Hence, CD44 deficiency led to decreased influx of neutrophils regardless of comparable levels in chemotactic factors expressed in the kidney. The reduced influx of neutrophils was associated with preserved renal function and morphology. Adoptive transfer experiments of labeled neutrophils revealed that endothelial CD44 rather than neutrophil CD44 mediates neutrophil migration. Activation of neutrophils increased cell-surface expression of hyaluronic acid (HA). Altogether, a novel mechanism in the recruitment of neutrophils that involves interaction of endothelial CD44 and neutrophil HA was found. Either blocking endothelial CD44 or removal of neutrophil HA decreased rolling and adhesion of neutrophils. Administration of anti-CD44 to mice reduced the influx of neutrophils into the postischemic tissue, associated with renal function preservation. Therefore, anti-CD44-based therapies may contribute to prevent or reduce renal I/R injury. PMID- 15901766 TI - Double-blind comparison of full and partial anemia correction in incident hemodialysis patients without symptomatic heart disease. AB - It is unclear whether physiologic hemoglobin targets lead to cardiac benefit in incident hemodialysis patients without symptomatic heart disease and left ventricular dilation. In this randomized, double-blind study, lower (9.5 to 11.5 g/dl) and higher (13.5 to 14.5 g/dl) hemoglobin targets were generated with epoetin alpha over 24 wk and maintained for an additional 72 wk. Major eligibility criteria included recent hemodialysis initiation and absence of symptomatic cardiac disease and left ventricular dilation. The primary outcome measure was left ventricular volume index (LVVI). The study enrolled 596 patients. Mean age, duration of dialysis therapy, baseline predialysis hemoglobin, and LVVI were 50.8 yr, 0.8 yr, 11.0 g/dl, and 69 ml/m2, respectively; 18% had diabetic nephropathy. Mean hemoglobin levels in the higher and lower target groups were 13.3 and 10.9 g/dl, respectively, at 24 wk. Percentage changes in LVVI between baseline and last value were similar (7.6% in the higher and 8.3% in the lower target group) as were the changes in left ventricular mass index (16.8 versus 14.2%). For the secondary outcomes, the only between-group difference was an improved SF-36 Vitality score in the higher versus the lower target group (1.21 versus -2.31; P = 0.036). Overall adverse event rates were similar in both target groups; higher (P < 0.05) rates of skeletal pain, surgery, and dizziness were seen in the lower target group, and headache and cerebrovascular events were seen in the higher target group. Normalization of hemoglobin in incident hemodialysis patients does not have a beneficial effect on cardiac structure, compared with partial correction. PMID- 15901767 TI - RhoGTPase activation is a key step in renal epithelial mesenchymal transdifferentiation. AB - ESRD is characterized by an interstitial infiltrate of inflammatory cells in association with tubular atrophy, epithelial mesenchymal transdifferentiation (EMT), and interstitial fibrosis. Human proximal tubular epithelial cells (HK2 cells) undergo EMT in response to activated PBMC conditioned medium (aPBMC-CM), showing acquisition of a fibroblastoid morphology, increased fibronectin-EDA (EDA) expression, loss of junctional E-cadherin localization, and cytokeratin 19 (K19) expression. The signaling pathway(s) that regulates EMT in response to aPBMC-CM is not well understood. This study shows that aPBMC-CM induces a rapid activation of RhoA, Rac1, and Cdc42 activity in HK2 cells from 15 min to 48 h. Moreover, infection with adenovirus expressing constitutively active RhoA, Rac1, and Cdc42 significantly increased the expression of EDA and downregulated expression of E-cadherin and K19. Dominant negative RhoA expression suppressed aPBMC-CM-induced upregulation of EDA but did not restore the expression of E cadherin and K19. Constitutively active RhoA activated the Rho kinase and its downstream effectors, whereas constitutively active Rac1 and Cdc42 activated the P21-activated protein kinase in epithelial cells. In further experiments, HK2 cells were treated with toxin B, exoenzyme C3, Y-27632, and HA1077. These strategies, inhibiting the Rho/Rho kinase pathway, as well as the Rac1/Cdc42/P21 activated protein kinase pathway, blocked transdifferentiation of HK2 cells in response to aPBMC-CM. To conclude, these results indicate that aPBMC-CM activates RhoA, Rac1, and Cdc42 and their downstream effectors, leading to HK2 cells undergoing transdifferentiation. Therefore, activation of small RhoGTPases is a key step in the mechanism of EMT and likely to be a contributor to tubulointerstitial fibrosis. PMID- 15901768 TI - Endoplasmic reticulum stress-associated caspase 12 mediates cisplatin-induced LLC PK1 cell apoptosis. AB - Reactive oxygen metabolites are important mediators in cisplatin-induced apoptosis in renal tubular epithelial cells (LLC-PK1). Mitochondria have been implicated to play a principal role in cisplatin-induced apoptosis. Caspase 12, an endoplasmic reticulum (ER)-specific caspase, participates in apoptosis under ER stress. Cytochrome P450 system is crucial to the generation of reactive oxygen metabolites and is present at high concentration in the ER. The direct role of caspase 12 in any model of renal injury has not previously been described. In this study, cleavage of procaspase 12 preceded that of caspases 3 and 9 after cisplatin treatment of LLC-PK1 cells. The active form of caspase 8 was not detected throughout the course of study. Preincubation of the LLC-PK1 cells with the caspase 9 inhibitor did not attenuate caspase 3 activation and provided no significant protection. Caspase 3 inhibitor provided only modest protection against cisplatin-induced apoptosis. LLC-PK1 cells that were transfected with anti-caspase 12 antibody significantly attenuated cisplatin-induced apoptosis. Taken together, these data indicate that caspase 12 plays a pivotal role in cisplatin-induced apoptosis. It is proposed that the oxidative stress that results from the interaction of cisplatin with the ER cytochrome P450 leads to activation of procaspase 12, resulting in apoptosis. PMID- 15901769 TI - Membrane proteinase 3 expression in patients with Wegener's granulomatosis and in human hematopoietic stem cell-derived neutrophils. AB - A large membrane proteinase 3 (mPR3)-positive neutrophil subset (mPR3high) is a risk for Wegener's granulomatosis (WG). The relationship between mPR3 expression and clinical manifestations was investigated in 81 WG patients and mPR3 expression was studied in CD34+ stem cell-derived human neutrophils. The mPR3high neutrophil percentage correlated with renal function, anemia, and albumin at the time of presentation. The mPR3high neutrophil percentage and renal failure severity correlated directly after 5 yr. For elucidating mechanisms that govern mPR3 expression, studies were conducted to determine whether the genetic information that governs mPR3 expression resides within the neutrophils, even without stimuli possibly related to disease. CD34+ hematopoietic stem cells were differentiated to neutrophils, and their mPR3 expression was determined. A two step amplification/differentiation protocol was used to differentiate human CD34+ hematopoietic stem cells into neutrophils with G-CSF. The cells progressively expressed the neutrophil surface markers CD66b, CD35, and CD11b. The ferricytochrome C assay demonstrated a strong respiratory burst at day 14 in response to PMA but none at day 0. Intracellular PR3 was detectable from day 4 by Western blotting. An increasing percentage of a mPR3-positive neutrophil subset became detectable by flow cytometry, whereas a second subset remained negative, consistent with a bimodal expression. Finally, human PR3-anti-neutrophil cytoplasmic autoantibodies induced a stronger respiratory burst, compared with human control IgG in stem cell-derived neutrophils. Taken together, these studies underscore the clinical importance of the WG mPR3 phenotype. The surface mPR3 on resting cells is probably genetically determined rather than being dictated by external factors. PMID- 15901770 TI - Mutually exclusive glomerular innervation by two distinct types of olfactory sensory neurons revealed in transgenic zebrafish. AB - The olfactory epithelium of fish contains two major types of olfactory sensory neurons (OSNs) that are distinct morphologically (ciliated vs microvillous) and possibly functionally. Here, we found that these OSNs express different sets of signal transduction machineries: the ciliated OSNs express OR-type odorant receptors, cyclic nucleotide-gated channel A2 subunit, and olfactory marker protein (OMP), whereas the microvillous OSNs express V2R-type receptors and transient receptor potential channel C2 (TRPC2). To visualize patterns of axonal projection from the two types of OSNs to the olfactory bulb (OB), we generated transgenic zebrafish in which spectrally distinct fluorescent proteins are expressed in the ciliated and microvillous OSNs under the control of OMP and TRPC2 gene promoters, respectively. An observation of whole-mount OB in adult double-transgenic zebrafish revealed that the ciliated OSNs project axons mostly to the dorsal and medial regions of the OB, whereas the microvillous OSNs project axons to the lateral region of the OB. A careful histological examination of OB sections clarified that the axons from the two distinct types of OSNs target different glomeruli in a mutually exclusive manner. This segregation is already established at very early developmental stages in zebrafish embryos. These findings clearly demonstrate the relationships among cell morphology, molecular signatures, and axonal terminations of the two distinct types of OSNs and suggest that the two segregated neural pathways are responsible for coding and processing of different types of odor information in the zebrafish olfactory system. PMID- 15901771 TI - Camguk/CASK enhances Ether-a-go-go potassium current by a phosphorylation dependent mechanism. AB - Signaling complexes are essential for the modulation of excitability within restricted neuronal compartments. Adaptor proteins are the scaffold around which signaling complexes are organized. Here, we demonstrate that the Camguk (CMG)/CASK adaptor protein functionally modulates Drosophila Ether-a-go-go (EAG) potassium channels. Coexpression of CMG with EAG in Xenopus oocytes results in a more than twofold average increase in EAG whole-cell conductance. This effect depends on EAG-T787, the residue phosphorylated by calcium- and calmodulin dependent protein kinase II (Wang et al., 2002). CMG coimmunoprecipitates with wild-type and EAG-T787A channels, indicating that T787, although necessary for the effect of CMG on EAG current, is not required for the formation of the EAG CMG complex. Both CMG and phosphorylation of T787 increase the surface expression of EAG channels, and in COS-7 cells, EAG recruits CMG to the plasma membrane. The interaction of EAG with CMG requires a noncanonical Src homology 3-binding site beginning at position R1037 of the EAG sequence. Mutation of basic residues, but not neighboring prolines, prevents binding and prevents the increase in EAG conductance. Our findings demonstrate that membrane-associated guanylate kinase adaptor proteins can modulate ion channel function; in the case of CMG, this occurs via an increase in the surface expression and phosphorylation of the EAG channel. PMID- 15901772 TI - Regulation of activin mRNA and Smad2 phosphorylation by antidepressant treatment in the rat brain: effects in behavioral models. AB - Activin is a member of the transforming growth factor-beta family that is involved in cell differentiation, hormone secretion, and regulation of neuron survival. The cellular responses to activin are mediated by phosphorylation of a downstream target, Smad2. The current study examines the influence of chronic electroconvulsive seizures (ECSs), as well as chemical antidepressants, on the expression of activin betaA and the phosphorylation of Smad2 in the rat hippocampus and frontal cortex. Chronic ECSs (10 d) resulted in a significant increase in activin betaA mRNA expression and Smad2 phosphorylation in both the hippocampus and frontal cortex. Chronic fluoxetine did not influence activin betaA expression, but fluoxetine as well as desipramine did increase Smad2 phosphorylation in the frontal cortex. The functional significance of increased activin was further tested by examining the effects of activin infusions into the hippocampus on a behavioral model of depression, the forced swim test (FST). A single bilateral infusion of activin A or activin B into the dentate gyrus of the hippocampus produced an antidepressant-like effect in the FST that was comparable in magnitude with fluoxetine. In contrast, infusion of the activin antagonist inhibin A did not influence behavior but blocked the effect of activin A. The results suggest that regulation of activin and Smad signaling may contribute to the actions of antidepressant treatment and may represent novel targets for antidepressant drug development. PMID- 15901773 TI - Tissue inhibitor of metalloproteinase-2 promotes neuronal differentiation by acting as an anti-mitogenic signal. AB - Although traditionally recognized for maintaining extracellular matrix integrity during morphogenesis, the function of matrix metallo-proteinases (MMPs) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), in the mature nervous system is essentially unknown. Here, we report that TIMP-2 induces pheochromocytoma PC12 cell-cycle arrest via regulation of cell-cycle regulatory proteins, resulting in differentiation and neurite outgrowth. TIMP-2 decreases cyclins B and D expression and increases p21Cip expression. Furthermore, TIMP-2 promotes cell differentiation via activation of the cAMP/Rap1/ERK (extracellular signal-regulated kinase) pathway. Expression of dominant-negative Rap1 blocks TIMP-2-mediated neurite outgrowth. Both the cell-cycle arrest and neurite outgrowth induced by TIMP-2 was independent of MMP inhibitory activity. Consistent with the PC12 cell data, primary cultures of TIMP-2 knock-out cerebral cortical neurons exhibit significantly reduced neurite length, which is rescued by TIMP-2. These in vitro results were corroborated in vivo. TIMP-2 deletion causes a delay in neuronal differentiation, as demonstrated by the persistence of nestin-positive progenitors in the neocortical ventricular zone. The interaction of TIMP-2 with alpha3beta1 integrin in the cerebral cortex suggests that TIMP-2 promotes neuronal differentiation and maintains mitotic quiescence in an MMP independent manner through integrin activation. The identification of molecules responsible for neuronal quiescence has significant implications for the ability of the adult brain to generate new neurons in response to injury and neurological disorders, such as Alzheimer's and Parkinson's diseases. PMID- 15901775 TI - Large-field visual motion directly induces an involuntary rapid manual following response. AB - Recent neuroscience studies have been concerned with how aimed movements are generated on the basis of target localization. However, visual information from the surroundings as well as from the target can influence arm motor control, in a manner similar to known effects in postural and ocular motor control. Here, we show an ultra-fast manual motor response directly induced by a large-field visual motion. This rapid response aided reaction when the subject moved his hand in the direction of visual motion, suggesting assistive visually evoked manual control during postural movement. The latency of muscle activity generating this response was as short as that of the ocular following responses to the visual motion. Abrupt visual motion entrained arm movement without affecting perceptual target localization, and the degrees of motion coherence and speed of the visual stimulus modulated this arm response. This visuomotor behavior was still observed when the visual motion was confined to the "follow-through" phase of a hitting movement, in which no target existed. An analysis of the arm movements suggests that the hitting follow through made by the subject is not a part of a reaching movement. Moreover, the arm response was systematically modulated by hand bias forces, suggesting that it results from a reflexive control mechanism. We therefore propose that its mechanism is radically distinct from motor control for aimed movements to a target. Rather, in an analogy with reflexive eye movement stabilizing a retinal image, we consider that this mechanism regulates arm movements in parallel with voluntary motor control. PMID- 15901774 TI - Modulation of presynaptic Ca2+ entry by AMPA receptors at individual GABAergic synapses in the cerebellum. AB - Cerebellar Purkinje cells (PCs) receive GABAergic input that undergoes powerful retrograde modulation by presynaptic cannabinoid and glutamate receptors. Here we examine a distinct modulatory mechanism at these synapses, which does not require postsynaptic depolarization and acts via presynaptic AMPA receptors. We find that this mechanism operates mainly in the somatic vicinity of PCs in which large boutons of basket cell axons form synapses on the PC soma. We use fast confocal microscopy and detailed kinetic modeling to estimate that, in these boutons, an action potential opens 100-200 Ca2+ channels, eliciting a brief 3-5 microM transient, followed by a longer-term, 15-30 nM rise of free Ca2+ (above the resting level of approximately 100 nM). Brief activation of local AMPA receptors suppresses Ca2+ entry (probably by silencing 20-40 P/Q-type channels) in a subgroup of terminals that tend to show a higher dynamic range of Ca2+ signaling. The results provide the first quantitative description of presynaptic Ca2+ kinetics and its modulation by AMPA receptor activation (most likely via a glutamate spillover-mediated mechanism) at identified GABAergic synapses. PMID- 15901776 TI - Selectivity of canary HVC neurons for the bird's own song: modulation by photoperiodic conditions. AB - To what extent seasonal factors modify the neuronal functional properties within the nuclei of the avian song system remains an open question. In adult songbirds, neurons of the song premotor nucleus HVC (used as a proper name) exhibit selective responses for the bird's own song (BOS). Here we examine whether, outside the breeding season, when songs are less stereotyped, HVC neurons of male canaries still respond selectively to the BOS produced during this period. In an initial experiment, single-unit recordings (n = 114) revealed that the neuronal selectivity for the current BOS was attenuated in males exposed to a short-day photoperiod (typical of the nonbreeding season) compared with that found in males exposed to a long-day photoperiod. In long-day conditions, 35% of the cells responded to the BOS, whereas only 12% did in short-day conditions; there were four times more selective cells (d' > 1) in long-day than in short-day conditions. To determine whether these effects were the consequence of differences in acoustic features between breeding and nonbreeding songs, neurons (n = 72) recorded in short-day conditions were tested with both a short-day BOS and a long-day BOS. A low percentage of neurons exhibited responses to short-day or to long-day BOS (11% for each song). Responses of putative interneurons (spike duration < 0.4 ms) and of putative relay cells were similarly attenuated by the short-day conditions. These results strongly suggest that, in canary, rather than being a fixed property, the selectivity for the BOS moves along a continuum and peaks when the day length mimics the breeding conditions. PMID- 15901777 TI - Adaptation of sensory neurons to hyalectin and decorin proteoglycans. AB - Proteoglycans are abundantly expressed in the pathways of developing and regenerating neurons, yet the responses of neurons to specific proteoglycans are not well characterized. We have shown previously that one chondroitin sulfate proteoglycan (CSPG), aggrecan, is potently inhibitory to sensory axon extension in short-term assays and that over time, embryonic neurons adapt to aggrecan mediated inhibition through the transcriptional upregulation of integrin expression (Condic et al., 1999). Here, we have compared the response of embryonic sensory neurons to structurally distinct CSPGs that belong to either the hyalectin (or lectican) family of large, aggregating proteoglycans or the decorin (or small leucine-rich proteoglycan) family of smaller proteoglycans. Both of these structurally diverse proteoglycan families are expressed in developing embryos and inhibit outgrowth of embryonic sensory neurons in short term cultures. These results document a previously uncharacterized inhibitory function for the decorin-family proteoglycan biglycan. Interestingly, embryonic neurons adapt to these diverse proteoglycans over time. Adaptation is associated with upregulation of select integrin alpha subunits in a proteoglycan-specific manner. Overexpression of specific integrin alpha subunits improves neuronal regeneration on some but not all decorin-family CSPGs, suggesting that neurons adapt to inhibition mediated by closely related proteoglycans using distinct mechanisms. Our findings indicate that CSPGs with diverse core proteins and distinct numbers of chondroitin sulfate substitution sites mediate a similar response in sensory neurons, suggesting that increased integrin expression may be an effective means of promoting axonal regeneration in the presence of diverse inhibitory proteoglycan species in vivo. PMID- 15901778 TI - Regulation of NMDA receptors by neuregulin signaling in prefrontal cortex. AB - Recent linkage studies have identified a significant association of the neuregulin gene with schizophrenia, but how neuregulin is involved in schizophrenia is primarily unknown. Aberrant NMDA receptor functions have been implicated in the pathophysiology of schizophrenia. Therefore, we hypothesize that neuregulin, which is present in glutamatergic synaptic vesicles, may affect NMDA receptor functions via actions on its ErbB receptors enriched in postsynaptic densities, hence participating in emotional regulation and cognitive processes that are impaired in schizophrenia. To test this, we examined the regulation of NMDA receptor currents by neuregulin signaling pathways in prefrontal cortex (PFC), a prominent area affected in schizophrenia. We found that bath perfusion of neuregulin significantly reduced whole-cell NMDA receptor currents in acutely isolated and cultured PFC pyramidal neurons and decreased NMDA receptor-mediated EPSCs in PFC slices. The effect of neuregulin was mainly blocked by application of the ErbB receptor tyrosine kinase inhibitor, phospholipase C (PLC) inhibitor, IP3 receptor (IP3R) antagonist, or Ca2+ chelators. The neuregulin regulation of NMDA receptor currents was also markedly attenuated in cultured neurons transfected with mutant forms of Ras or a dominant negative form of MEK1 (mitogen-activated protein kinase kinase 1). Moreover, the neuregulin effect was prevented by agents that stabilize or disrupt actin polymerization but not by agents that interfere with microtubule assembly. Furthermore, neuregulin treatment increased the abundance of internalized NMDA receptors in cultured PFC neurons, which was also sensitive to agents affecting actin cytoskeleton. Together, our study suggests that both PLC/IP3R/Ca2+ and Ras/MEK/ERK (extracellular signal-regulated kinase) signaling pathways are involved in the neuregulin-induced reduction of NMDA receptor currents, which is likely through enhancing NR1 internalization via an actin-dependent mechanism. PMID- 15901779 TI - Variation of input-output properties along the somatodendritic axis of pyramidal neurons. AB - The firing evoked by injection of simulated barrages of EPSCs into the proximal dendrite of layer 5 pyramidal neurons is greater than when comparable inputs are injected into the soma. This boosting is mediated by dendritic Na+ conductances. However, the presence of other active conductances in the dendrites, some of which are nonuniformly distributed, suggests that the degree of boosting may differ along the somatodendritic axis. Here, we injected EPSC barrages at the soma and at the proximal, middle, and distal segments of the apical dendrite and measured boosting of subthreshold and suprathreshold responses. We found that although boosting was maintained throughout the apical dendrite, the degree of boosting changed nonmonotonically with distance from the soma. Boosting dipped in the middle dendritic segments as a result of the deactivation of the hyperpolarization-activated cation current, Ih, but increased in the distal dendrites as a result of the activation of Ca2+ conductances. In the distal dendrites, EPSC barrages evoked repetitive bursts of action potentials, and the bursting pattern changed systematically with the magnitude of the input barrages. The quantitative changes in boosting along the somatodendritic axis suggest that inputs from different classes of presynaptic cells are weighted differently, depending on the location of the synaptic contacts. Moreover, the tight coupling between burst characteristics and stimulus parameters indicate that the distal dendrites can support a coding scheme that is different from that at sites closer to the soma, consistent with the notion of a separate dendritic integration site. PMID- 15901780 TI - Regulation of alpha-synuclein expression in limbic and motor brain regions of morphine-treated mice. AB - Chronic exposure to opiates produces dependence and addiction, which may result from neuroadaptations in the dopaminergic reward pathway and its target brain regions. The neuronal protein alpha-synuclein has been implicated in neuronal plasticity and proposed to serve as a negative regulator of dopamine neurotransmission. Thus, alpha-synuclein could mediate some effects of opiates in the brain. The present study investigated the influence of acute and chronic morphine administration on alpha-synuclein mRNA and protein expression in the brains of mice. Downregulation of alpha-synuclein mRNA was observed in the basolateral amygdala, dorsal striatum, nucleus accumbens, and ventral tegmental area of mice withdrawn from chronic morphine treatment. The changes were the most pronounced after longer periods of withdrawal (48 h). In contrast, levels of alpha-synuclein protein, as assessed by Western blotting, were significantly increased in the amygdala and striatum/accumbens (but not in the mesencephalon) of morphine-withdrawn mice. In both brain regions, levels of alpha-synuclein were elevated for as long as 2 weeks after treatment cessation. Because alpha synuclein is a presynaptic protein, the detected opposite changes in its mRNA and protein levels are likely to take place in different populations of projection neurons whose somata are in different brain areas. Axonal localization of alpha synuclein was confirmed by immunofluorescent labeling. An attempt to identify postsynaptic neurons innervated by alpha-synuclein-containing axon terminals revealed their selective apposition to calbindin D28K-negative projection neurons in the basolateral amygdala. The observed changes in alpha-synuclein levels are discussed in connection with their putative role in mediating suppression of dopaminergic neurotransmission during opiate withdrawal. PMID- 15901781 TI - Multisensory integration of dynamic faces and voices in rhesus monkey auditory cortex. AB - In the social world, multiple sensory channels are used concurrently to facilitate communication. Among human and nonhuman primates, faces and voices are the primary means of transmitting social signals (Adolphs, 2003; Ghazanfar and Santos, 2004). Primates recognize the correspondence between species-specific facial and vocal expressions (Massaro, 1998; Ghazanfar and Logothetis, 2003; Izumi and Kojima, 2004), and these visual and auditory channels can be integrated into unified percepts to enhance detection and discrimination. Where and how such communication signals are integrated at the neural level are poorly understood. In particular, it is unclear what role "unimodal" sensory areas, such as the auditory cortex, may play. We recorded local field potential activity, the signal that best correlates with human imaging and event-related potential signals, in both the core and lateral belt regions of the auditory cortex in awake behaving rhesus monkeys while they viewed vocalizing conspecifics. We demonstrate unequivocally that the primate auditory cortex integrates facial and vocal signals through enhancement and suppression of field potentials in both the core and lateral belt regions. The majority of these multisensory responses were specific to face/voice integration, and the lateral belt region shows a greater frequency of multisensory integration than the core region. These multisensory processes in the auditory cortex likely occur via reciprocal interactions with the superior temporal sulcus. PMID- 15901783 TI - Tuft calcium spikes in accessory olfactory bulb mitral cells. AB - The mammalian accessory olfactory system is critical for the detection and identification of pheromones and the representation of complex stimuli including sex, genetic relatedness, and individual identity. Mitral cells, the principal cells of the accessory olfactory bulb (AOB), receive monosynaptic input from the sensory periphery and already show highly specific response properties, firing selectively for combinations of genetic markers and gender-specific cues. Vomeronasal sensory neuron axons form synapses onto distal tuft-like branches of mitral cell primary dendrites. We have studied dendritic excitability and synaptic integration in AOB mitral cell dendrites, and we show that dendrites of accessory olfactory bulb mitral cells support action potential propagation and can fire regenerative spike-like events that are likely to contribute to the integration of inputs to these cells. These tuft spikes may be important for the specificity of AOB mitral cell responses. PMID- 15901784 TI - Endogenous kappa-opioid receptor systems regulate mesoaccumbal dopamine dynamics and vulnerability to cocaine. AB - Genetic and pharmacological approaches were used to examine kappa-opioid receptor (KOR-1) regulation of dopamine (DA) dynamics in the nucleus accumbens and vulnerability to cocaine. Microdialysis revealed that basal DA release and DA extraction fraction (Ed), an indirect measure of DA uptake, are enhanced in KOR-1 knock-out mice. Analysis of DA uptake revealed a decreased Km but unchanged Vmax in knock-outs. Knock-out mice exhibited an augmented locomotor response to cocaine, which did not differ from that of wild-types administered a behavioral sensitizing cocaine treatment. The ability of cocaine to increase DA was enhanced in knock-outs, whereas c-fos induction was decreased. Although repeated cocaine administration to wild types produced behavioral sensitization, knock-outs exhibited no additional enhancement of behavior. Administration of the long acting KOR antagonist nor-binaltorphimine to wild-type mice increased DA dynamics. However, the effects varied with the duration of KOR-1 blockade. Basal DA release was increased whereas Ed was unaltered after 1 h blockade. After 24 h, release and Ed were increased. The behavioral and neurochemical effects of cocaine were enhanced at both time points. These data demonstrate the existence of an endogenous KOR-1 system that tonically inhibits mesoaccumbal DA neurotransmission. Its loss induces neuroadaptations characteristic of "cocaine sensitized" animals, indicating a critical role of KOR-1 in attenuating responsiveness to cocaine. The increased DA uptake after pharmacological inactivation or gene deletion highlights the plasticity of mesoaccumbal DA neurons and suggests that loss of KOR-1 and the resultant disinhibition of DA neurons trigger short- and long-term DA transporter adaptations that maintain normal DA levels, despite enhanced release. PMID- 15901782 TI - Mesocortical dopamine neurons operate in distinct temporal domains using multimodal signaling. AB - In vivo extracellular recording studies have traditionally shown that dopamine (DA) transiently inhibits prefrontal cortex (PFC) neurons, yet recent biophysical measurements in vitro indicate that DA enhances the evoked excitability of PFC neurons for prolonged periods. Moreover, although DA neurons apparently encode stimulus salience by transient alterations in firing, the temporal properties of the PFC DA signal associated with various behaviors is often extraordinarily prolonged. The present study used in vivo electrophysiological and electrochemical measures to show that the mesocortical system produces a fast non DA-mediated postsynaptic response in the PFC that appears to be initiated by glutamate. In contrast, short burst stimulation of mesocortical DA neurons that produced transient (<4 s) DA release in the PFC caused a simultaneous reduction in spontaneous firing (consistent with extracellular in vivo recordings) and a form of DA-induced potentiation in which evoked firing was increased for tens of minutes (consistent with in vitro measurements). We suggest that the mesocortical system might transmit fast signals about reward or salience via corelease of glutamate, whereas the simultaneous prolonged DA-mediated modulation of firing biases the long-term processing dynamics of PFC networks. PMID- 15901785 TI - Effect of catechol-O-methyltransferase val158met genotype on attentional control. AB - The cingulate cortex is richly innervated by dopaminergic projections and plays a critical role in attentional control (AC). Evidence indicates that dopamine enhances the neurophysiological signal-to-noise ratio and that dopaminergic tone in the frontal cortex is critically dependent on catechol-O-methyltransferase (COMT). A functional polymorphism (val158met) in the COMT gene accounts for some of the individual variability in executive function mediated by the dorsolateral prefrontal cortex. We explored the effect of this genetic polymorphism on cingulate engagement during a novel AC task. We found that the COMT val158met polymorphism also affects the function of the cingulate during AC. Individuals homozygous for the high-activity valine ("val") allele show greater activity and poorer performance than val/methionine ("met") heterozygotes, who in turn show greater activity and poorer performance than individuals homozygous for the low activity met allele, and these effects are most evident at the highest demand for AC. These results indicate that met allele load and presumably enhanced dopaminergic tone improve the "efficiency" of local circuit processing within the cingulate cortex and thereby its function during AC. PMID- 15901786 TI - Physiological maturation of photoreceptors depends on the voltage-gated sodium channel NaV1.6 (Scn8a). AB - Voltage-gated sodium channels (VGSCs) ensure the saltatory propagation of action potentials along axons by acting as signal amplifiers at the nodes of Ranvier. In the retina, activity mediated by VGSCs is important for the refinement of the retinotectal map. Here, we conducted a full-field electroretinogram (ERG) study on mice null for the sodium channel NaV1.6. Interestingly, the light-activated hyperpolarization of photoreceptor cells (the a-wave) and the major "downstream" components of the ERG, the b-wave and the oscillatory potentials, are markedly reduced and delayed in these mice. The functional deficit was not associated with any morphological abnormality. We demonstrate that Scn8a is expressed in the ganglion and inner nuclear layers and at low levels in the outer nuclear layer beginning shortly before the observed ERG deficit. Together, our data reveal a previously unappreciated role for VGSCs in the physiological maturation of photoreceptors. PMID- 15901787 TI - Molecular determinants of KCNQ (Kv7) K+ channel sensitivity to the anticonvulsant retigabine. AB - Epilepsy is caused by an electrical hyperexcitability in the CNS. Because K+ channels are critical for establishing and stabilizing the resting potential of neurons, a loss of K+ channels could support neuronal hyperexcitability. Indeed, benign familial neonatal convulsions, an autosomal dominant epilepsy of infancy, is caused by mutations in KCNQ2 or KCNQ3 K+ channel genes. Because these channels contribute to the native muscarinic-sensitive K+ current (M current) that regulates excitability of numerous types of neurons, KCNQ (Kv7) channel activators would be effective in epilepsy treatment. A compound exhibiting anticonvulsant activity in animal seizure models is retigabine. It specifically acts on the neuronally expressed KCNQ2-KCNQ5 (Kv7.2-Kv7.5) channels, whereas KCNQ1 (Kv7.1) is not affected. Using the differential sensitivity of KCNQ3 and KCNQ1 to retigabine, we constructed chimeras to identify minimal segments required for sensitivity to the drug. We identified a single tryptophan residue within the S5 segment of KCNQ3 and also KCNQ2, KCNQ4, and KCNQ5 as crucial for the effect of retigabine. Furthermore, heteromeric KCNQ channels comprising KCNQ2 and KCNQ1 transmembrane domains (attributable to transfer of assembly properties from KCNQ3 to KCNQ1) are retigabine insensitive. Transfer of the tryptophan into the KCNQ1 scaffold resulted in retigabine-sensitive heteromers, suggesting that the tryptophan is necessary in all KCNQ subunits forming a functional tetramer to confer drug sensitivity. PMID- 15901788 TI - The functional divide for primary reinforcement of D-amphetamine lies between the medial and lateral ventral striatum: is the division of the accumbens core, shell, and olfactory tubercle valid? AB - When projection analyses placed the nucleus accumbens and olfactory tubercle in the striatal system, functional links between these sites began to emerge. The accumbens has been implicated in the rewarding effects of psychomotor stimulants, whereas recent work suggests that the medial accumbens shell and medial olfactory tubercle mediate the rewarding effects of cocaine. Interestingly, anatomical evidence suggests that medial portions of the shell and tubercle receive afferents from common zones in a number of regions. Here, we report results suggesting that the current division of the ventral striatum into the accumbens core and shell and the olfactory tubercle does not reflect the functional organization for amphetamine reward. Rats quickly learned to self-administer D amphetamine into the medial shell or medial tubercle, whereas they failed to learn to do so into the accumbens core, ventral shell, or lateral tubercle. Our results suggest that primary reinforcement of amphetamine is mediated via the medial portion of the ventral striatum. Thus, the medial shell and medial tubercle are more functionally related than the medial and ventral shell or the medial and lateral tubercle. The current core-shell-tubercle scheme should be reconsidered in light of recent anatomical data and these functional findings. PMID- 15901789 TI - Estradiol activates group I and II metabotropic glutamate receptor signaling, leading to opposing influences on cAMP response element-binding protein. AB - In addition to mediating sexual maturation and reproduction through stimulation of classical intracellular receptors that bind DNA and regulate gene expression, estradiol is also thought to influence various brain functions by acting on receptors localized to the neuronal membrane surface. Many intracellular signaling pathways and modulatory proteins are affected by estradiol via this unconventional route, including regulation of the transcription factor cAMP response element-binding protein (CREB). However, the mechanisms by which estradiol acts at the membrane surface are poorly understood. Because both estradiol and CREB have been implicated in regulating learning and memory, we characterized the effects of estradiol on this transcription factor in cultured rat hippocampal neurons. Within minutes of administration, estradiol triggered mitogen-activated protein kinase (MAPK)-dependent CREB phosphorylation in unstimulated neurons. Furthermore, after brief depolarization, estradiol attenuated L-type calcium channel-mediated CREB phosphorylation. Thus, estradiol exhibited both positive and negative influences on CREB activity. These effects of estradiol were sex specific and traced to membrane-localized estrogen receptors that stimulated group I and II metabotropic glutamate receptor (mGluR) signaling. Activation of estrogen receptor alpha (ERalpha) led to mGluR1a signaling, triggering CREB phosphorylation through phospholipase C regulation of MAPK. In addition, estradiol stimulation of ERalpha or ERbeta triggered mGluR2/3 signaling, decreasing L-type calcium channel-mediated CREB phosphorylation. These results not only characterize estradiol regulation of CREB but also provide two putative signaling mechanisms that may account for many of the unexplained observations regarding the influence of estradiol on nervous system function. PMID- 15901790 TI - Pallidal origin of GABA release within the substantia nigra pars reticulata during high-frequency stimulation of the subthalamic nucleus. AB - High-frequency stimulation of the subthalamic nucleus (HFS-STN) is an effective treatment for alleviating the motor symptoms of parkinsonian patients. However, the neurochemical basis of its effects remains unknown. We showed previously that 1 h of HFS-STN in normal rats increases extracellular glutamate (Glu) level in the output nuclei of the STN, the globus pallidus (GP), and the substantia nigra pars reticulata (SNr), consistent with an increase in the activity of STN neurons. HFS-STN also increases GABA levels in the SNr, but the origin of this increase is unclear. We investigated the effectiveness of HFS-STN for improving Parkinson's disease symptoms, using intracerebral microdialysis to determine the extracellular Glu and GABA levels of the GP and SNr in response to HFS-STN in anesthetized hemiparkinsonian rats [6-hydroxydopamine lesion of the substantia nigra pars compacta (SNc)]. Basal levels of Glu and GABA in the GP and SNr were significantly higher in hemiparkinsonian than in intact rats. HFS-STN did not affect extracellular Glu level in the SNr of hemiparkinsonian rats but doubled the level of GABA. Ibotenic acid lesion of the GP abolished the increase in GABA levels in the SNr induced by HFS-STN in SNc-lesioned rats. These results provide neurochemical confirmation of the hyperactivity of the STN after dopaminergic denervation and suggest that the therapeutic effects of HFS-STN may result partly from the stimulation of pallidonigral fibers, thereby revealing a potential role for pallidal GABA in the inhibition of basal ganglial output structures during HFS-STN. PMID- 15901791 TI - Postadolescent changes in regional cerebral protein synthesis: an in vivo study in the FMR1 null mouse. AB - Methylation-induced transcriptional silencing of the fragile X mental retardation 1 (Fmr1) gene leads to absence of the gene product, fragile X mental retardation protein (FMRP), and consequently fragile X syndrome (FrX), an X-linked inherited form of mental retardation. Absence of FMRP in Fmr1 null mice imparts some characteristics of the FrX phenotype, but the precise role of FMRP in neuronal function remains unknown. FMRP is an RNA-binding protein that has been shown to suppress translation of certain mRNAs in vitro. We applied the quantitative autoradiographic L-[1-14C]leucine method to the in vivo determination of regional rates of cerebral protein synthesis (rCPS) in adult wild-type (WT) and Fmr1 null mice at 4 and 6 months of age. Our results show a substantial decrease in rCPS in all brain regions examined between the ages of 4 and 6 months in both WT and Fmr1 null mice. Superimposed on the age-dependent decline in rCPS, we demonstrate a regionally selective elevation in rCPS in Fmr1 null mice. Our results suggest that the process of synaptic pruning during young adulthood may be reflected in decreased rCPS. Our findings support the hypothesis that FMRP is a suppressor of translation in brain in vivo. PMID- 15901792 TI - Dynamic activation of cystic fibrosis transmembrane conductance regulator by type 3 and type 4D phosphodiesterase inhibitors. AB - The diseases of cystic fibrosis, chronic obstructive pulmonary disease (COPD), and chronic bronchitis are characterized by mucus-congested and inflamed airways. Anti-inflammatory agents that can simultaneously restore or enhance mucociliary clearance through cystic fibrosis transmembrane conductance regulator (CFTR) activation may represent new therapeutics in their treatment. Herein, we report the activation of CFTR-mediated chloride secretion by phosphodiesterase (PDE) 4 inhibitors in T84 monolayer using (125)I anion as tracer. In the absence of forskolin, the iodide secretion was insensitive to PDE4 inhibitor L-826,141 [4-[2 (3,4-bis-difluoromethoxyphenyl)-2-[4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2 yl)phenyl]-ethyl]-3-methylpyridine-1-oxide], roflumilast, or to PDE3 inhibitor trequinsin. However, these inhibitors potently augmented iodide secretion after forskolin stimulation, with efficacy coupled to the activation states of adenylyl cyclase. The iodide secretion from PDE3 or PDE4 inhibition was characterized at first by a prolonged efflux duration, followed by progressively elevated peak efflux rates at higher inhibitor concentrations. Paralleled with an increased phosphor-cAMP response element-binding protein formation, the CFTR activation dissociated from a global cAMP elevation and was blocked by H89 [N-[2-((p bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide]. 2-(4-Fluorophenoxy)-N [(1S)-1-(4-methoxyphenyl)ethyl]nicotinamide, a stereoselective PDE4D inhibitor, augmented iodide efflux more efficiently than its less potent (R)-isomer. The peak efflux from maximal PDE4 and PDE3 inhibition matched that from full adenylyl cyclase activation. These data suggest that PDE3 and PDE4 (mainly PDE4D) form the major cAMP diffusion barrier in T84 cells to ensure a compartmentalized CFTR signaling. Together with their potent anti-inflammatory properties, the potentially enhanced airway mucociliary clearance from CFTR activation may have contributed to the efficacy of PDE4 inhibitors in COPD and asthmatic patients. PDE4 inhibitors may represent new opportunities to combat cystic fibrosis and other respiratory diseases in future. PMID- 15901793 TI - Antianalgesia: stereoselective action of dextro-morphine over levo-morphine on glia in the mouse spinal cord. AB - We have previously shown that the naturally occurring levo-morphine at a subanalgesic picomolar dose pretreated i.t. induces antianalgesia against levo morphine-produced antinociception. We now report that the synthetic stereo enantiomer dextro-morphine, even at an extremely low femtomolar dose, induces antianalgesia against levo-morphine-produced antinociception using the tail-flick (TF) test in male CD-1 mice. Intrathecal pretreatment with dextro-morphine (33 fmol) time-dependently attenuated the i.t. levo-morphine-produced TF inhibition for 4 h and returned to the preinjection control level at 24 h. Intrathecal pretreatment with dextro-morphine (0.3-33 fmol), which injected alone did not affect the baseline TF latency, dose-dependently attenuated the TF inhibition produced by i.t.-administered levo-morphine (3.0 nmol). The ED(50) value for dextro-morphine to induce antianalgesia was estimated to be 1.07 fmol, which is 71,000-fold more potent than the ED(50) value of levo-morphine, indicating the high stereoselective action of dextro-morphine over levo-morphine for the induction of antianalgesia. Like levo-morphine, the dextro-morphine-induced antianalgesia against levo-morphine-produced TF inhibition was dose-dependently blocked by the nonopioid dextro-naloxone and its stereo-enantiomer levo-naloxone, a nonselective mu-opioid receptor antagonist. The antianalgesia induced by levo morphine and dextro-morphine is reversed by the pretreatment with the glial inhibitor propentofylline (3.3-65 nmol), indicating that the antianalgesia is mediated by glial stimulation. The findings strongly indicate that the antianalgesia induced by levo-morphine and dextro-morphine is mediated by the stimulation of a novel nonopioid receptor on glial cells. PMID- 15901794 TI - (+)-Norfenfluramine-induced arterial contraction is not dependent on endogenous 5 hydroxytryptamine or 5-hydroxytryptamine transporter. AB - (+)-Norfenfluramine, the major metabolite of fenfluramine, causes vasoconstriction dependence on the 5-hydroxytryptamine (5-HT)(2A) receptor in rat. (+)-Norfenfluramine was reported as a 5-hydroxytryptamine transporter (5 HTT) substrate and 5-HT releaser. Because the arterial 5-HTT exists and is functional in the rat, we hypothesized that (+)-norfenfluramine causes vasoconstriction by releasing 5-HT from vascular smooth muscle via 5-HTT. The released 5-HT, in turn, activates the 5-HT(2A) receptor. Isometric contractility experiments showed that (+)-norfenfluramine-induced mouse aortic contraction was reduced by the 5-HTT inhibitor fluoxetine (1 micriM) but not by fluvoxamine (1 microM). Tryptophan hydroxylase (TPH)-deficient (Tph1-/-) mice lack peripheral 5 HT. (+)-Norfenfluramine (10 nM-100 microM)-contracted aorta from wild-type and Tph1-/- mice with equivalent potency (-log EC(50) [M], wild type = 5.73 +/- 0.02, Tph1-/- = 5.62 +/- 0.09), and these contractions were inhibited by the 5-HT(2A) receptor antagonist ketanserin (3 nM) by a similar magnitude in aorta from wild type and Tph1-/- mice (wild type = 19.4, Tph1-/- = 15.4-fold rightward shift versus control), as did fluoxetine (1 microM) (wild type = 22.4, Tph1-/- = 28.8 fold rightward shift versus control). To further test the role of 5-HTT in (+) norfenfluramine-induced aortic contraction, the 5-HTT-targeted mutation mouse was used. (+)-Norfenfluramine induced similar aortic contraction in wild-type and 5 HTT-targeted mutation mice, and these contractions were inhibited by fluoxetine (1 microM). Thus, (+)-norfenfluramine vasoconstriction is not dependent on 5-HTT mediated release of endogenous 5-HT but by activating membrane 5-HT(2A) receptors directly. Understanding of the mechanism by which (+)-norfenfluramine induces vasoconstriction is important to characterize and understand the function of the serotonergic system in peripheral arterial vasculature. PMID- 15901795 TI - Pharmacological implications of two distinct mechanisms of interaction of memantine with N-methyl-D-aspartate-gated channels. AB - Unlike other N-methyl-D-aspartate receptor (NMDAR) antagonists, clinical trials have shown that memantine is clinically tolerated and effective in the treatment of Alzheimer's disease. The mechanism for memantine tolerability, however, remains contentious but may be partly explained by its uncompetitive antagonism. The specific site of memantine block in the NMDAR channel interacts with magnesium and is assumed to be at or near a narrow constriction representing the channel selectivity filter. A second, very low-affinity site of memantine action has also been reported. Here, using mutational analysis and substituted cysteine accessibility methods on recombinant NR1/NR2A NMDARs expressed in Xenopus oocytes, we precisely localize both the specific and second memantine-blocking sites. Intriguingly, memantine interacts with its specific blocking site in the same fashion as intracellular rather than extracellular Mg(2+). Thus, the N-site asparagine (N) in the M2 region of the NR1 subunit represents the dominant site for uncompetitive antagonism by memantine. The N and N + 1 site asparagines in NR2A produce strong electrostatic interactions with memantine. In contrast, the second (superficial) memantine-blocking site, located at the extracellular vestibule of the channel, appears to be nonspecific and overlaps the site occupied by the nonspecific pore blocker hexamethonium. Residues in the post-M3 segment of the NR1 subunit are not directly involved in memantine binding. The distinct patterns of interaction and the relative degree of affinity of memantine for these two binding sites contribute to the drug's excellent pharmacological profile of clinical tolerability. In the future, these parameters should be considered in searching for improved neuroprotective agents in this class. PMID- 15901796 TI - Bile salt export pump (BSEP/ABCB11) can transport a nonbile acid substrate, pravastatin. AB - Pravastatin is a well known 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor. Cumulative studies have shown that pravastatin is taken up into hepatocytes by the organic anion transporting polypeptide family transporters and excreted into the bile as an intact form by multidrug resistance-associated protein 2 (MRP2). It is generally accepted that the bile salt export pump (BSEP/ABCB11) mainly transports bile acids and plays an indispensable role in their biliary excretion. Interestingly, we found that BSEP could accept pravastatin as a substrate. Significant ATP-dependent uptake of pravastatin by human BSEP (hBSEP)- and rat BSEP (rBsep)-expressing membrane vesicles was observed, and the ratio of the uptake activity of pravastatin to that of taurocholic acid (TCA) by hBSEP was 3.3 fold higher than that by rBsep. The K(m) value of pravastatin for hBSEP was 124 muM. A mutual inhibition study between TCA and pravastatin revealed that they competitively interact with hBSEP. Several statins inhibited the hBSEP- and rBsep mediated uptake of TCA; however, the specific uptake of other statins (cerivastatin, fluvastatin, and pitavastatin) by hBSEP and rBSEP was not detected. The inhibitory effects of hydrophilic statins (pravastatin and rosuvastatin) on the uptake of TCA by BSEP were relatively lower than those of lipophilic statins. These data suggest that BSEP may be partly involved in the biliary excretion of pravastatin in both rats and humans. PMID- 15901797 TI - Amino acid ester prodrugs of 2-bromo-5,6-dichloro-1-(beta-D ribofuranosyl)benzimidazole enhance metabolic stability in vitro and in vivo. AB - 2-Bromo-5,6-dichloro-1-(beta-d-ribofuranosyl)benzimidazole (BDCRB) is a potent and selective inhibitor of human cytomegalovirus (HCMV), but it lacks clinical utility due to rapid in vivo metabolism. We hypothesized that amino acid ester prodrugs of BDCRB may enhance both in vitro potency and systemic exposure of BDCRB through evasion of BDCRB-metabolizing enzymes. To this end, eight different amino acid prodrugs of BDCRB were tested for N-glycosidic bond stability, ester bond stability, Caco-2 cell uptake, antiviral activity, and cytotoxicity. The prodrugs were resistant to metabolism by BDCRB-metabolizing enzymes, and ester bond cleavage was rate-limiting in metabolite formation from prodrug. Thus, BDCRB metabolism could be controlled by the selection of promoiety. In HCMV plaque formation assays, l-Asp-BDCRB exhibited 3-fold greater selectivity than BDCRB for inhibition of HCMV replication. This potent and selective antiviral activity in addition to favorable stability profile made l-Asp-BDCRB an excellent candidate for in vivo assessment and pharmacokinetic comparison with BDCRB. In addition to rapid absorption and sufficient prodrug activation after oral administration to mice, l-Asp-BDCRB exhibited a 5-fold greater half-life than BDCRB. Furthermore, the sum of area under the concentration-time profile (AUC)(BDCRB) and AUC(prodrug) after l-Asp-BDCRB administration was roughly 3-fold greater than AUC(BDCRB) after BDCRB administration, suggesting that a reservoir of prodrug was delivered in addition to parent drug. Overall, these findings demonstrate that amino acid prodrugs of BDCRB exhibit evasion of metabolizing enzymes (i.e., bioevasion) in vitro and provide a modular approach for translating this in vitro stability into enhanced in vivo delivery of BDCRB. PMID- 15901798 TI - Development of polyethylene glycol-conjugated poly-S-nitrosated serum albumin, a novel S-Nitrosothiol for prolonged delivery of nitric oxide in the blood circulation in vivo. AB - S-Nitrosothiols are an interesting class of nitric oxide (NO) donors used for the treatment of circulation disorders. In this study, we developed a novel macromolecular NO donor in which 10 NO molecules were covalently bound to polyethylene glycol (PEG)-conjugated bovine serum albumin (BSA) through S nitrosothiol linkages (PEG-poly SNO-BSA). Intermolecular disulfide linkages possibly formed during the introduction of thiol groups to BSA were prevented in PEG-poly SNO-BSA. Electron spin resonance study indicated that PEG-poly SNO-BSA does release the NO radical in the blood circulation in vivo. The area under the concentration-time curve of (111)In-PEG-poly N-succinimidyl S-acetylthioacetate (SATA)-BSA, the carrier part of PEG-poly SNO-BSA, was 1.7 times greater than that of (111)In-BSA after intravenous injection in mice. After intravenous injection in rats at an equivalent NO dose (3 micromol of NO per kilogram), the duration of reduction in the blood pressure was 2.3 to 3.7 times longer in PEG-poly SNO-BSA than in classic S-nitrosothiols such as S-nitroso-N-acetyl penicillamine, S nitrosoglutathione, and NO-BSA. The release half-life of NO from PEG-poly SNO-BSA was 11 to 108 times longer than those of the classic S-nitrosothiols examined, and this slow release rate of NO would explain the sustained reduction in the blood pressure after intravenous injection of PEG-poly SNO-BSA in rats. No cross tolerance between PEG-poly SNO-BSA and nitroglycerin was also observed. These findings indicate that the novel S-nitrosothiol PEG-poly SNO-BSA is a promising compound that exhibits unique characteristics of sustained release of NO in the blood circulation in vivo, which would be beneficial for the treatment of circulation disorders. PMID- 15901799 TI - Mechanism of the vascular angiotensin II/alpha2-adrenoceptor interaction. AB - alpha(2)-Adrenoceptors potentiate vascular responses to angiotensin II. The goal of this study was to test the hypothesis that the phospholipase C (PLC)/protein kinase C (PKC)/c-src/phosphatidylinositol 3-kinase (PI3K) pathway contributes to the vascular angiotensin II/alpha(2)-adrenoceptor interaction. In rats in vivo, intrarenal infusions of angiotensin II (10 ng/kg/min) increased renal vascular resistance by 5.8 +/- 0.5 units, and this response was enhanced (p < 0.05) to 9.1 +/- 1.2 units by UK-14,304 [5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6 quinoxalinamine; 3 microg/kg/min; alpha(2)-adrenoceptor agonist]. Intrarenal infusions of U-73122 [1-[6-[[(17beta)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino] hexyl]-1H-pyrrole-2,5-dione; 3 microg/min; PLC inhibitor], GF109203X [bisindolylmaleimide I; 10 microg/min; PKC inhibitor], CGP77675 [1-(2-{4-[4-amino 5-(3-methoxyphenyl)pyrrolo[2,3-d]pyrimidin-7-yl]phenyl}ethyl)piperidin-4-ol; 5 microg/min; c-src inhibitor], and wortmannin (1 microg/min; PI3K inhibitor) abolished the angiotensin II/alpha(2)-adrenoceptor interaction. In isolated perfused rat kidneys, angiotensin II (0.3, 1, and 3 nM) increased perfusion pressure (by 15 +/- 8, 39 +/- 4, and 93 +/- 9 mm Hg, respectively), and UK-14,304 (1 microM) potentiated these responses (to 36 +/- 4, 67 +/- 7, and 135 +/- 17 mm Hg, respectively). This angiotensin II/alpha(2)-adrenoceptor interaction was abolished by U-73122 (10 microM), GF109203X (3 microM), CGP77675 (5 microM), and wortmannin (0.2 microM). Preglomerular microvascular smooth muscle cells expressed phospholipase (PLC)-beta(2), PLC-beta(3), c-src, phospho(tyrosine 416) c-src, and PI3K. In these cells, angiotensin II (0.1 microM) and UK-14,304 (1 microM) per se did not increase phospho-c-src; however, the combination of angiotensin II plus UK-14,304 doubled phospho-c-src, and this interaction was abolished by U-73122 (10 microM) and GF109203X (3 microM). In conclusion, the PLC/PKC/c-src/PI3K pathway may contribute importantly to the interaction between alpha(2)-adrenoceptors and angiotensin II on renal vascular resistance. PMID- 15901800 TI - Identification of the hepatic efflux transporters of organic anions using double transfected Madin-Darby canine kidney II cells expressing human organic anion transporting polypeptide 1B1 (OATP1B1)/multidrug resistance-associated protein 2, OATP1B1/multidrug resistance 1, and OATP1B1/breast cancer resistance protein. AB - Until recently, it was generally believed that the transport of various organic anions across the bile canalicular membrane was mainly mediated by multidrug resistance-associated protein 2 (MRP2/ABCC2). However, a number of new reports have shown that some organic anions are also substrates of multidrug resistance 1 (MDR1/ABCB1) and/or breast cancer resistance protein (BCRP/ABCG2), implying MDR1 and BCRP could also be involved in the biliary excretion of organic anions in humans. In the present study, we constructed new double-transfected Madin-Darby canine kidney II (MDCKII) cells expressing organic anion-transporting polypeptide 1B1 (OATP1B1)/MDR1 and OATP1B1/BCRP, and we investigated the transcellular transport of four kinds of organic anions, estradiol-17beta-d-glucuronide (EG), estrone-3-sulfate (ES), pravastatin (PRA), and cerivastatin (CER), to identify which efflux transporters mediate the biliary excretion of compounds using double transfected cells. We observed the vectorial transport of EG and ES in all the double transfectants. MRP2 showed the highest efflux clearance of EG among these efflux transporters, whereas BCRP-mediated clearance of ES was the highest in these double transfectants. In addition, two kinds of 3-hydroxy-3-methylglutaryl CoA reductase inhibitors, CER and PRA, were also substrates of all these efflux transporters. The rank order of the efflux clearance of PRA mediated by each transporter was the same as that of EG, whereas the contribution of MDR1 to the efflux of CER was relatively greater than for PRA. This experimental system is very useful for identifying which transporters are involved in the biliary excretion of organic anions that cannot easily penetrate the plasma membrane. PMID- 15901801 TI - Mitogen-activated protein kinase phosphorylation in the rostral ventrolateral medulla plays a key role in imidazoline (i1)-receptor-mediated hypotension. AB - Our previous study showed that rilmenidine, a selective I(1)-imidazoline receptor agonist, enhanced the phosphorylation of mitogen-activated protein kinase (MAPK)(p42/44), via the phosphatidylcholine-specific phospholipase C pathway in the pheochromocytoma cell line (PC12). In the present study, we tested the hypothesis that enhancement of MAPK phosphorylation in the rostral ventrolateral medulla (RVLM) contributes to the hypotensive response elicited by I(1)-receptor activation in vivo. Systemic rilmenidine (600 microg/kg i.v.) elicited hypotension and bradycardia along with significant elevation in MAPK(p42/44), detected by immunohistochemistry, in RVLM neurons. To obtain conclusive evidence that the latter response was I(1)-receptor-mediated, similar hypotensive responses were elicited by intracisternal (i.c.) rilmenidine (25 microg/rat) or the highly selective alpha(2)-agonist alpha-methylnorepinephrine (4 microg/rat). An increase in RVLM MAPK(p42/44) occurred only after rilmenidine. Furthermore, pretreatment with efaroxan (0.15 microg/rat i.c.), a selective I(1)-imidazoline receptor antagonist, or with PD98059 (2'-amino-3'-methoxyflavone) (5 microg/rat i.c.), a selective extracellular signal-regulated kinase 1/2 inhibitor, significantly attenuated the hypotensive response and the elevation in RVLM MAPK(p42/44) elicited by i.c. rilmenidine. The findings suggest that MAPK phosphorylation in the RVLM contributes to the hypotensive response induced by I(1)-receptor activation and presents in vivo evidence that distinguishes the neuronal responses triggered by the I(1)-receptor from those triggered by the alpha(2)-adrenergic receptor. PMID- 15901802 TI - Delineation of human peptide transporter 1 (hPepT1)-mediated uptake and transport of substrates with varying transporter affinities utilizing stably transfected hPepT1/Madin-Darby canine kidney clones and Caco-2 cells. AB - In the present investigation, the uptake and transport kinetics of valacyclovir (VACV), 5-aminolevulinic acid (5-ALA), and benzylpenicillin (BENZ) were studied in stably transfected Madin-Darby canine kidney (MDCK)/human peptide transporter 1 (hPepT1)-V5&His clonal cell lines expressing varying levels of epitope-tagged hPepT1 protein (low, medium, and high expression) and in Caco-2 cells to delineate hPepT1-mediated transport kinetics. These compounds were selected due to the fact that they are known PepT1 substrates, yet also have affinity for other transporters. Caco-2 cells, traditionally used for studying peptide-based drug transport, were included for comparison purposes. The time, pH, sodium, and concentration dependence of cellular uptake and permeability were measured using mock, clonal hPepT1-MDCK, and Caco-2 cells. A pH-dependent effect was observed in the hPepT1-expressing clones and Caco-2 cells, with an increase of 1.96-, 1.84-, and 2.05-fold for VACV, 5-ALA, and BENZ uptake, respectively, at pH 6 versus 7.4 in the high-expressing hPepT1 cells. BENZ uptake was significantly decreased in Caco-2 and MDCK cells in Na(+)-depleted buffer, whereas VACV uptake only decreased in Caco-2 cells. Concentration-dependent uptake studies in the mock corrected hPepT1-MDCK and Caco-2 cells demonstrated hPepT1 affinity ranking of VACV > 5-ALA > BENZ. The apical-to-basal apparent permeability coefficient (P(app)) values of VACV, 5-ALA, and BENZ in mock-corrected hPepT1-MDCK cells showed solely hPepT1-mediated transport in contrast to Caco-2 cells. Lower K(m) values and higher P(app) in Caco-2 cells compared with hPepT1-MDCK cells suggested the involvement of multiple transporters in Caco-2 cells. Thus, hPepT1 MDCK cells corrected for endogenous transporter expression may be a more appropriate model for screening compounds for their affinity to hPepT1. PMID- 15901803 TI - Contributions of the mitogen-activated protein kinase and protein kinase C cascades in spatial learning and memory mediated by the nucleus accumbens. AB - Several studies have reported a role for the nucleus accumbens (NAcc) in learning and memory. Specifically, NAcc seems to function as a neural bridge for the translation of corticolimbic information to the motor system mediating locomotor learning, but the signaling mechanisms involved in this striatal learning await further investigation. The present experiments investigated the role of the mitogen-activated protein kinase (MAPK) and protein kinase C (PKC) cascades within the NAcc of Long-Evans rats in a food-search spatial learning task (FSSLT). First, we used immunoblotting to examine changes in MAPK p42/p44 phosphorylation within the NAcc in the acquisition phase of the FSSLT. Second, we examined the effect on the acquisition and retention phases in the FSSLT of pretraining intra-accumbal microinjections of the MAPK [U0126; 1,4-diamino-2,3 dicyano-1,4-bis(2-aminophynyltio)butadiene, 1 microg/side] or PKC [GF109203X; bisindolylmaleimide or 1-(3-dimethylaminopropyl)-indol-3-yl]-3-(indol-3-yl) maleimide, 0.5 ng/side] inhibitors (four training sessions; one session/day). Third, the potential coupling of PKC and MAPK signaling pathways in the NAcc in spatial learning was studied using microinjections of GF109203X, radioactive activity assays, and immunoblotting. Results showed that 1) MAPK p42/p44 phosphorylation is augmented within the NAcc after spatial learning, 2) MAPK and PKC inhibition caused differential deficits in the acquisition and formation of spatial memories, and 3) inhibition of PKC activity by GF109203X caused a reduction in MAPKs phosphorylation in the NAcc in an early stage of the acquisition phase. Overall, these findings suggest that NAcc-PKC and -MAPK play important roles in spatial learning and that MAPKs phosphorylation seems to be mediated through the activation of the PKC signaling pathway. PMID- 15901804 TI - Age-dependent methamphetamine-induced alterations in vesicular monoamine transporter-2 function: implications for neurotoxicity. AB - Tens of thousands of adolescents and young adults have used illicit methamphetamine. This is of concern since its high-dose administration causes persistent dopaminergic deficits in adult animal models. The effects in adolescents are less studied. In adult rodents, toxic effects of methamphetamine may result partly from aberrant cytosolic dopamine accumulation and subsequent reactive oxygen species formation. The vesicular monoamine transporter-2 (VMAT-2) sequesters cytoplasmic dopamine into synaptic vesicles for storage and perhaps protection against dopamine-associated oxidative consequences. Accordingly, aberrant VMAT-2 function may contribute to the methamphetamine-induced persistent dopaminergic deficits. Hence, this study examined effects of methamphetamine on VMAT-2 in adolescent (postnatal day 40) and young adult (postnatal day 90) rats. Results revealed that high-dose methamphetamine treatment caused greater acute (within 1 h) decreases in vesicular dopamine uptake in postnatal day 90 versus 40 rats, as determined in a nonmembrane-associated subcellular fraction. Greater basal levels of VMAT-2 at postnatal day 90 versus 40 in this purified fraction seemed to contribute to the larger effect. Basal tissue dopamine content was also greater in postnatal day 90 versus 40 rats. In addition, postnatal day 90 rats were more susceptible to methamphetamine-induced persistent dopaminergic deficits as assessed by measuring VMAT-2 activity and dopamine content 7 days after treatment, even if drug doses were adjusted for age-related pharmacokinetic differences. Together, these data demonstrate dynamic changes in VMAT-2 susceptibility to methamphetamine as a function of development. Implications with regard to methamphetamine-induced dopaminergic deficits, as well as dopamine associated neurodegenerative disorders such as Parkinson's disease, are discussed. PMID- 15901805 TI - The endocannabinoid noladin ether acts as a full agonist at human CB2 cannabinoid receptors. AB - Noladin ether (NE) is a putative endogenously occurring cannabinoid demonstrating agonist activity at CB1 receptors. Because of reported selective affinity for CB1 receptors, the pharmacological actions of NE at CB2 receptors have not been examined. Therefore, the purpose of this study was to characterize the binding and functional properties of NE at human CB2 receptors stably expressed in Chinese hamster ovary (CHO) cells as well as in HL-60 cells, which express CB2 receptors endogenously. Surprisingly, in transfected CHO cells, NE exhibits a relatively high nanomolar affinity for CB2 receptors (K(i) = 480 nM), comparable to that observed for the endocannabinoid 2-arachidonoyl glycerol (2-AG) (K(i) = 1016 nM). Furthermore, NE activates G proteins and inhibits the intracellular effector adenylyl cyclase with equivalent efficacy relative to the full cannabinoid agonists 2-AG and CP 55,940 (CP) [(-)-cis-3-[2-hydroxy-4-(1,1 dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl) cyclohexanol]. The rank order of potency for G protein activation and effector regulation by the three agonists is similar to their apparent affinity for CB2 receptors; CP > NE > or = 2-AG. Regulation of adenylyl cyclase activity by all agonists is inhibited by pertussis toxin pretreatment or by coincubation with AM630 [6-iodo-2-methyl-1-[2-(4 morpholinyl)ethyl]-1H-indol-3-yl](4-methoxyphenyl)-methanone], a CB2 antagonist. Chronic treatment with NE or CP results in CB2 receptor desensitization and down regulation. All agonists also inhibit adenylyl cyclase activity in HL-60 cells. Together, these data indicate that NE acts as a full agonist at human CB2 receptors and thus might have important physiological functions at peripheral cannabinoid receptors. PMID- 15901806 TI - Binding profile of the endogenous novel heptapeptide Met-enkephalin-Gly-tyr in zebrafish and rat brain. AB - Zebrafish is considered a model organism, not only for the study of the biological functions of vertebrates but also as a tool to analyze the effects of some drugs or toxic agents. Five opioid precursor genes homologous to the mammalian opioid propeptide genes have recently been identified; one of these, the zebrafish proenkephalin, codes a novel heptapeptide, the Met-enkephalin-Gly Tyr (MEGY). To analyze the pharmacological properties of this novel ligand, we have labeled it with tritium ([(3)H]MEGY). In addition, we have also synthesized two analogs: (d-Ala(2))-MEGY (Y-d-Ala-GFMGY) and (d-Ala(2), Val(5))-MEGY (Y-d-Ala GFVGY). The binding profile of these three agents has been studied in zebrafish and rat brain membranes. [(3)H]MEGY presents one binding site in zebrafish, as well as in rat brain membranes, although it shows a slight higher affinity in zebrafish brain. The observed saturable binding is displaced by naloxone, thus confirming the opioid nature of the binding sites. Competition binding assays indicate that the methionine residue is essential for high-affinity binding of MEGY and probably of other peptidic agonists in zebrafish, whereas the change of a Gly for a d-Ala does not dramatically affect the ligand affinity. Our results show that the percentage of [(3)H]MEGY displaced by all the ligands studied is higher than 100%, thus inferring that naloxone (used to determine nonspecific binding) does not bind to all the sites labeled by [(3)H]MEGY. Therefore, we can deduct that some of the MEGY binding sites should not be considered classical opioid sites. PMID- 15901807 TI - Metatarsus primus elevatus in hallux rigidus: fact or fiction? AB - Two hundred seventy-five lateral weightbearing radiographs of isolated pathology were reviewed and stratified into hallux rigidus (n = 100), hallux valgus (n = 75), plantar fasciitis (n = 50), and Morton's neuroma (n = 50) groups. The patient population consisted of healthy individuals with no history of foot trauma or surgery. The first to second metatarsal head elevation, Seiberg index, first to second sagittal intermetatarsal angle, first to fifth metatarsal head distance, and hallux equinus angle were measured in each population. Statistically significant differences were found between the hallux valgus, plantar fasciitis, and Morton's neuroma populations and the hallux rigidus population, which showed greater elevation of the first metatarsal relative to the second for each radiographic measurement technique. In the hallux rigidus population, there was a statistically significant difference between grade II and grades I and III regarding the first to fifth metatarsal head distance (greater in grade II) and the hallux equinus angle (lower in grade II). A review of the literature and comparison with historical controls reveals that metatarsus primus elevatus exists in hallux rigidus and is greater than that found in hallux valgus, plantar fasciitis, and Morton's neuroma groups. PMID- 15901808 TI - Cimetidine as a first-line therapy for pedal verruca: eight-year retrospective analysis. AB - Can cimetidine therapy effectively stimulate the body's immune response against warts? Several clinicians have anecdotally reported success using cimetidine against warts. Previous double-blind studies comparing cimetidine with placebo therapy have failed to statistically and scientifically corroborate those results. Between 1995 and 2002, 216 patients underwent an isolated course of oral cimetidine therapy for verruca plantaris. Our treatment outcomes closely parallel those obtained by other researchers. Cimetidine may be used as a safe, effective, lone treatment modality for verruca in all age groups. PMID- 15901809 TI - Foot type and overuse injury in triathletes. AB - Abnormal foot morphology has been suggested to contribute to overuse injuries in athletes. This study investigated the relationship between foot type and injury incidence in a large sample of competitive triathletes not wearing foot orthoses during a 6-month retrospective analysis and a 10-week prospective cohort study. Foot alignment was measured using the Foot Posture Index and the Valgus Index, and participants were assigned to supinated, pronated, and normal foot-type groups. Overall, 131 triathletes sustained 155 injuries during the study. Generally, foot type was not a major risk factor for injury; however, there was a fourfold increased risk of overuse injury during the competition season in athletes with a supinated foot type. The results of this study show that triathletes with a supinated foot type are more likely to sustain an overuse injury. PMID- 15901810 TI - Evaluation of Young's modulus in Achilles tendons with diabetic neuroarthropathy. AB - The Achilles tendon of the patient with Charcot's foot neuroarthropathy has significantly altered physical properties compared with a normal tendon. Twenty nine Achilles tendons from patients with Charcot's foot (n = 20) and non Charcot's foot controls (n = 9) were loaded onto a biomechanical testing instrument. The biomechanical properties of the Charcot and control tendons were determined and the tendons were evaluated for differences in ultimate tensile strength and elasticity (Young's modulus). Biomechanical test data show that there is a significant difference in ultimate tensile strength and elasticity between tendons of patients with Charcot's foot and those of non-Charcot's controls. The term diabetic tendo Achillis equinus is introduced as a new finding in diabetic neuroarthropathy. PMID- 15901811 TI - The stance phase of walking during late pregnancy: temporospatial and ground reaction force variables. AB - The purpose of this study was to investigate temporospatial and ground reaction force variables in the stance phase of walking during late pregnancy. An eight camera motion-analysis system was used to record 13 pregnant women at 38 weeks' gestation and again 8 weeks after birth. In late pregnancy, there was a wider step width, and mediolateral ground reaction force tended to be increased in a medial direction. The center of pressure moved more medially initially and less anteriorly at 100% of stance in late pregnancy. The differences suggest that women may adapt their gait to maximize stability in the stance phase of walking and to control mediolateral motion. PMID- 15901812 TI - Maggot therapy in "lower-extremity hospice" wound care: fewer amputations and more antibiotic-free days. AB - We sought to assess, in a case-control model, the potential efficacy of maggot debridement therapy in 60 nonambulatory patients (mean +/- SD age, 72.2 +/- 6.8 years) with neuroischemic diabetic foot wounds (University of Texas grade C or D wounds below the malleoli) and peripheral vascular disease. Twenty-seven of these patients (45%) healed during 6 months of review. There was no significant difference in the proportion of patients healing in the maggot debridement therapy versus control group (57% versus 33%). Of patients who healed, time to healing was significantly shorter in the maggot therapy than in the control group (18.5 +/- 4.8 versus 22.4 +/- 4.4 weeks). Approximately one in five patients (22%) underwent a high-level (above-the-foot) amputation. Patients in the control group were three times as likely to undergo amputation (33% versus 10%). Although there was no significant difference in infection prevalence in patients undergoing maggot therapy versus controls (80% versus 60%), there were significantly more antibiotic-free days during follow-up in patients who received maggot therapy (126.8 +/- 30.3 versus 81.9 +/- 42.1 days). Maggot debridement therapy reduces short-term morbidity in nonambulatory patients with diabetic foot wounds. PMID- 15901813 TI - Utility of histopathologic analysis in the evaluation of onychomycosis. AB - Onychomycosis is a common problem seen in clinical practice. Given the differential diagnosis of dystrophic nails, it is helpful to obtain a definitive diagnosis of dermatophyte infection before initiation of antifungal therapy. Potassium hydroxide preparation and fungal culture, which are typically used in the diagnosis of these infections, often yield false-negative results. Recent studies have suggested that nail plate biopsy with periodic acid-Schiff stain may be a very sensitive technique for the diagnosis of onychomycosis. In this article, we review the literature on the utility of histopathologic analysis in the evaluation of onychomycosis. Many of these studies indicate that biopsy with periodic acid-Schiff is the most sensitive method for diagnosing onychomycosis. We propose that histopathologic examination is indicated if the results of other methods are negative and clinical suspicion is high; therefore, it is a useful complementary technique in the diagnosis of onychomycosis. PMID- 15901814 TI - The Bristol Foot Score: developing a patient-based foot-health measure. AB - We sought to develop a patient-centered foot-health assessment tool by conducting in-depth interviews, focus groups, and surveys of relevant patient groups. A total of 400 hospital- and community-based podiatric patients took part in the development of the Bristol Foot Score, which was refined from a 41-item self administered questionnaire to one containing 15 items. Podiatric patients easily understood the final questionnaire, and rates of completion were excellent. Overall reliability was high (Cronbach alpha = .9036), and application of the Bland and Altman technique suggested that the foot score produced stable measurements over time. Statistically significant differences were detected in scores before and after toenail surgery, indicating that the Bristol Foot Score is sensitive to change. A poor level of concordance was found between the Bristol Foot Score and a Chiropody Assessment Criteria Score routinely used by podiatrists to assess the need for podiatric care. The Bristol Foot Score reflects patients' perceptions of their own foot health, providing a useful additional tool for evaluating the efficacy of interventions and describing foot health within populations. PMID- 15901815 TI - Arch index as a predictor of pes planus: a comparative study of indigenous Kenyans and Tanzanians. AB - We determined the arch index of able-bodied indigenous Kenyan and Tanzanian individuals free of foot pain by using their dynamic footprints to classify the foot arch type and determine the prevalence of pes planus according to a previously described method. Males had a significantly higher arch index than females in both groups, and the prevalence of pes planus in Kenyans was 432 per 1,000 population, the highest ever documented and twice as high as that in Tanzanians (203 per 1,000 population). The arch index is useful in determining the prevalence of pes planus and possibly predicting pathologic foot conditions, and it may serve as an early warning sign of structural and functional defects of the foot in a given population. PMID- 15901816 TI - Hallucal sesamoid osteonecrosis: an overlooked cause of forefoot pain. AB - Four cases of osteonecrosis of hallucal sesamoids are reported here. Surgical excision of necrotic sesamoid tissue yielded satisfactory results, with the patients reporting no residual pain. Although it has not been frequently addressed in the literature, avascular necrosis of the sesamoid bones should be considered in the differential diagnosis of persistent forefoot pain. PMID- 15901817 TI - Percutaneous tendo Achillis lengthening to promote healing of diabetic plantar foot ulceration. AB - The etiology of ulcerations related to increased plantar pressure in patients with diabetes mellitus is complex but frequently includes a component of gastrocnemius soleus equinus. One viable treatment option is percutaneous tendo Achillis lengthening as a means of increasing dorsiflexory range of motion and decreasing forefoot shear forces. This article presents three case reports illustrating the importance of reducing plantar pressure as a crucial component of treatment of diabetic forefoot ulcerations. PMID- 15901818 TI - Focal tuberculous osteomyelitis of the calcaneus secondary to direct extension from an infected retrocalcaneal bursa. AB - We report a case of focal tuberculous involvement of the posterior margin of the calcaneus with preservation of the articular margin. The route of infection was direct extension through tuberculous retrocalcaneal bursitis, a rare and atypical pathogenesis. Magnetic resonance imaging was helpful in ruling out neoplasm and in limiting the diagnosis to an inflammatory infectious process. PMID- 15901819 TI - Cutaneous larva migrans: case report with current recommendations for treatment. AB - Cutaneous larva migrans is a common skin pathology that occurs in people who have recently visited tropical or subtropical climates. Given the ubiquity of this condition, the podiatric physician may encounter cutaneous larva migrans during clinical practice and should be cognizant of the presenting signs and typical patient history given in these cases. We describe the case of a 62-year-old man who presented with a pruritic, erythematous, serpiginous lesion on the dorsum of his left foot after having vacationed in Florida for several weeks. The patient was treated successfully with oral thiabendazole, 500 mg after meals 4 times daily for 5 days. PMID- 15901820 TI - Osteochondroma of the talar neck: a rare cause of callosity of the foot dorsum. AB - Osteochondroma is the most common benign bone tumor. It rarely affects rearfoot bones, and only a few cases of talar osteochondroma have been reported. We report a case of a solitary osteochondroma of the talus that presented as a painful callus on the anterior portion of the ankle that was refractory to dermatologic treatment. PMID- 15901821 TI - Accurate nomenclature for forefoot nerve entrapment: a historical perspective. AB - Current medical nomenclature is often based on the early history of the condition, when the true etiology of the disease or condition was not known. Sadly, this incorrect terminology can become inextricably woven into the lexicon of mainstream medicine. More important, when this is the case, the terminology itself can become integrated into current clinical decision making and ultimately into surgical intervention for the condition. "Morton's neuroma" is perhaps the most striking example of this nomenclature problem in foot and ankle surgery. We aimed to delineate the historical impetus for the terminology still being used today for this condition and to suggest appropriate terminology based on our current understanding of its pathogenesis. We concluded that this symptom complex should be given the diagnosis of nerve compression and be further distinguished by naming the involved nerve, such as compression of the interdigital nerve to the third web space or compression of the third common plantar digital nerve. Although the nomenclature becomes longer, the pathogenesis is correct, and treatment decisions can be made accordingly. PMID- 15901822 TI - An analysis of articles published in JAPMA in 2004. PMID- 15901823 TI - Reliability and validity of clinically assessing first-ray mobility of the foot. PMID- 15901824 TI - Isotopically coded cleavable cross-linker for studying protein-protein interaction and protein complexes. AB - An emerging approach for studying protein-protein interaction in complexes is the combination of chemical cross-linking and mass spectrometric analysis of the cross-linked peptides (cross-links) obtained after proteolysis of the complex. This approach, however, has several challenges and limitations, including the difficulty of detecting the cross-links, the potential interference from non informative "cross-linked peptides" (dead end and intrapeptide cross-links), and unambiguous identification of the cross-links by mass spectrometry. Thus, we have synthesized an isotopically coded ethylene glycol bis(succinimidylsuccinate) derivate (D12-EGS), which contains 12 deuterium atoms for easy detection of cross links when applied in a 1:1 mixture with its H12 counterpart and is also cleavable for releasing the cross-linked peptides allowing unambiguous identification by MS sequencing. Moreover, hydrolytic cleavage permits rapid distinguishing between different types of cross-links. Cleavage of a dead end cross-link produces a doublet with peaks 4.03 Da apart, with the lower peak appearing at a molecular mass 162 Da lower than the mass of the H12 form of the original cross-linked peptide. Cleavage of an intrapeptide cross-link leads to a doublet 8.05 Da apart and 62 Da lower than the molecular mass of the H12 form of the original cross-linked peptide. Cleavage of an interpeptide cross-link forms a pair of 4.03-Da doublets, with the lower mass member of each pair each shifted up from its unmodified molecular weight by 82 Da because of the attached portion of the cross-linker. All of this information has been incorporated into a software algorithm allowing automatic screening and detection of cross-links and cross link types in matrix-assisted laser desorption/ionization mass spectra. In summary, the ease of detection of these species through the use of an isotopically coded cleavable cross-linker and our software algorithm, followed by mass spectrometric sequencing of the cross-linked peptides after cleavage, has been shown to be a powerful tool for studies of multi-component protein complexes. PMID- 15901825 TI - Systematic analysis of the epidermal growth factor receptor by mass spectrometry reveals stimulation-dependent multisite phosphorylation. AB - Multisite phosphorylation of proteins is a general mechanism for modulation of protein function and molecular interactions. Definition of phosphorylation sites and elucidation of the functional interplay between multiple phosphorylated residues in proteins are, however, a major analytical challenge in current molecular cell biology and proteomic research. In the present study, we used mass spectrometry to determine the major phosphorylated residues of the human epidermal growth factor (EGF) receptor at various well defined cellular conditions. Activation of EGF receptor was achieved by several types of stimulation, i.e. by sodium pervanadate, EGF, and integrin-dependent adhesion. The contribution of cell-matrix adhesion was also determined by activating the EGF receptor by EGF in cells kept in suspension. We developed an analytical strategy that combined miniaturized sample preparation techniques and MALDI tandem mass spectrometry and determined a total of nine phosphorylation sites in the EGF receptor. We discovered one novel phosphorylation site (Ser967) and revealed constitutive phosphorylation of Thr669, Ser967, Ser1002, and Tyr1045 and stimulation-dependent differential phosphorylation of Tyr1068, Tyr1086, Ser1142, Tyr1148, and Tyr1173. The EGF receptor was purified from HeLa cells or ECV304 cells by immunoprecipitation and SDS-PAGE and then digested with trypsin. Phosphopeptides in the range of 0.8-3.7 kDa were recovered by combinations of IMAC, perfusion chromatography, and graphite powder chromatography and subsequently detected and sequenced by MALDI quadrupole time-of-flight tandem mass spectrometry. Two phosphorylation sites were detected in the peptide 1137GSHQISLDNPDYQQDFFPK1155; however, only Tyr1148 was phosphorylated upon EGF treatment; in contrast Ser1142 was only phosphorylated by integrin-dependent adhesion in the absence of EGF treatment, suggesting differential phosphorylation of this region by distinct stimuli. This MALDI MS/MS-based analytical approach demonstrates the feasibility of systematic analysis of signaling molecules by mass spectrometry and provides new insights into the dynamics of receptor signaling processes. PMID- 15901826 TI - Metabolic activation-related CD147-CD98 complex. AB - Cell surface CD147 protein promotes production of matrix metalloproteinases and hyaluronan, associates with monocarboxylate transporters and integrins, and is involved in reproductive, neural, inflammatory, and tumor functions. Here we combined covalent cross-linking, mass spectrometric protein identification, and co-immunoprecipitation to show selective CD147 association with three major types of transporters (CD98 heavy chain (CD98hc)-L-type amino acid transporter, ASCT2, and monocarboxylate transporters) as well as a regulator of cell proliferation (epithelial cell adhesion molecule). In the assembly of these multicomponent complexes, CD147 and CD98hc play a central organizing role. RNA interference knock-down experiments established a strong connection between CD147 and CD98hc expression and a strong positive association of CD147 (and CD98hc) with cell proliferation. As the CD147-CD98hc complex and proliferation diminished, AMP activated protein kinase (a cellular "fuel gauge") became activated, indicating a disturbance of cellular energy metabolism. Our data point to a CD147-CD98 cell surface supercomplex that plays a critical role in energy metabolism, likely by coordinating transport of lactate and amino acids. Furthermore we showed how covalent cross-linking, together with mass spectrometry, can be used to identify closely associated transmembrane proteins. This approach should also be applicable to many other types of transmembrane proteins besides those associated with CD98hc and CD147. PMID- 15901827 TI - Proteome analysis of the rice etioplast: metabolic and regulatory networks and novel protein functions. AB - We report an extensive proteome analysis of rice etioplasts, which were highly purified from dark-grown leaves by a novel protocol using Nycodenz density gradient centrifugation. Comparative protein profiling of different cell compartments from leaf tissue demonstrated the purity of the etioplast preparation by the absence of diagnostic marker proteins of other cell compartments. Systematic analysis of the etioplast proteome identified 240 unique proteins that provide new insights into heterotrophic plant metabolism and control of gene expression. They include several new proteins that were not previously known to localize to plastids. The etioplast proteins were compared with proteomes from Arabidopsis chloroplasts and plastid from tobacco Bright Yellow 2 cells. Together with computational structure analyses of proteins without functional annotations, this comparative proteome analysis revealed novel etioplast-specific proteins. These include components of the plastid gene expression machinery such as two RNA helicases, an RNase II-like hydrolytic exonuclease, and a site 2 protease-like metalloprotease all of which were not known previously to localize to the plastid and are indicative for so far unknown regulatory mechanisms of plastid gene expression. All etioplast protein identifications and related data were integrated into a data base that is freely available upon request. PMID- 15901828 TI - An immunoproteomic approach for identification of clinical biomarkers for monitoring disease: application to cystic fibrosis. AB - Circulating antibodies can be used to probe protein arrays of body fluids, prepared by two-dimensional gel electrophoresis, for antigenic biomarker detection. However, detected proteins, particularly low abundance antigens, often remain unidentifiable due to proteome complexity and limiting sample amounts. Using a novel enrichment approach exploiting patient antibodies for isolation of antigenic biomarkers, we demonstrate how immunoproteomic strategies can accelerate biomarker discovery. Application of this approach as a means of identifying biomarkers was demonstrated for cystic fibrosis (CF) lung disease by isolation and identification of inflammatory-associated autoantigens, including myeloperoxidase and calgranulin B from sputum of subjects with CF. The approach was also exploited for isolation of proteins expressed by the Pseudomonas aeruginosa strain PA01. Capture of PA01 antigens using circulating antibodies from CF subjects implicated in vivo expression of Pseudomonas proteins. All CF subjects screened, but not controls, were immunoreactive against immunocaptured Pseudomonas proteins, representing stress (GroES and ferric iron-binding protein HitA), immunosuppressive (thioredoxin), and alginate synthetase pathway (nucleoside-diphosphate kinase) proteins, implicating their clinical relevance as biomarkers of infection. PMID- 15901829 TI - Regulation of membrane localization of Sanpodo by lethal giant larvae and neuralized in asymmetrically dividing cells of Drosophila sensory organs. AB - In Drosophila, asymmetric division occurs during proliferation of neural precursors of the central and peripheral nervous system (PNS), where a membrane associated protein, Numb, is asymmetrically localized during cell division and is segregated to one of the two daughter cells (the pIIb cell) after mitosis. numb has been shown genetically to function as an antagonist of Notch signaling and also as a negative regulator of the membrane localization of Sanpodo, a four-pass transmembrane protein required for Notch signaling during asymmetric cell division in the CNS. Previously, we identified lethal giant larvae (lgl) as a gene required for numb-mediated inhibition of Notch in the adult PNS. In this study we show that Sanpodo is expressed in asymmetrically dividing precursor cells of the PNS and that Sanpodo internalization in the pIIb cell is dependent cytoskeletally associated Lgl. Lgl specifically regulates internalization of Sanpodo, likely through endocytosis, but is not required for the endocytosis Delta, which is a required step in the Notch-mediated cell fate decision during asymmetric cell division. Conversely, the E3 ubiquitin ligase neuralized is required for both Delta endocytosis and the internalization of Sanpodo. This study identifies a hitherto unreported role for Lgl as a regulator of Sanpodo during asymmetric cell division in the adult PNS. PMID- 15901830 TI - Induction of human NF-IL6beta by epidermal growth factor is mediated through the p38 signaling pathway and cAMP response element-binding protein activation in A431 cells. AB - The CCAAT/enhancer binding protein delta (C/EBPdelta, CRP3, CELF, NF-IL6beta) regulates gene expression and plays functional roles in many tissues, such as in acute phase response to inflammatory stimuli, adipocyte differentiation, and mammary epithelial cell growth control. In this study, we examined the expression of human C/EBPdelta (NF-IL6beta) gene by epidermal growth factor (EGF) stimulation in human epidermoid carcinoma A431 cells. NF-IL6beta was an immediate early gene activated by the EGF-induced signaling pathways in cells. By using 5' serial deletion reporter analysis, we showed that the region comprising the -347 to +9 base pairs was required for EGF response of the NF-IL6beta promoter. This region contains putative consensus binding sequences of Sp1 and cAMP response element-binding protein (CREB). The NF-IL6beta promoter activity induced by EGF was abolished by mutating the sequence of cAMP response element or Sp1 sites in the -347/+9 base pairs region. Both in vitro and in vivo DNA binding assay revealed that the CREB binding activity was low in EGF-starved cells, whereas it was induced within 30 min after EGF treatment of A431 cells. However, no change in Sp1 binding activity was found by EGF treatment. Moreover, the phosphatidylinositol 3 (PI3)-kinase inhibitor (wortmannin) and p38(MAPK) inhibitor (SB203580) inhibited the EGF-induced CREB phosphorylation and the expression of NF-IL6beta gene in cells. We also demonstrated that CREB was involved in regulating the NF-IL6beta gene transcriptional activity mediated by p38(MAPK). Our results suggested that PI3-kinase/p38(MAPK)/CREB pathway contributed to the EGF activation of NF-IL6beta gene expression. PMID- 15901831 TI - Impaired Akt activity down-modulation, caspase-3 activation, and apoptosis in cells expressing a caspase-resistant mutant of RasGAP at position 157. AB - RasGAP bears two caspase-3 cleavage sites that are used sequentially as caspase activity increases in cells. When caspase-3 is mildly activated, RasGAP is first cleaved at position 455. This leads to the production of an N-terminal fragment, called fragment N, that activates the Ras-PI3K-Akt pathway and that promotes cell survival. At higher caspase activity, RasGAP is further cleaved at position 157 generating two small N-terminal fragments named N1 and N2. We have now determined the contribution of this second cleavage event in the regulation of apoptosis using cells in which the wild-type RasGAP gene has been replaced by a cDNA encoding a RasGAP mutant that cannot be cleaved at position 157. Our results show that cleavage of fragment N at position 157 leads to a marked reduction in Akt activity. This is accompanied by efficient processing of caspase-3 that favors cell death in response to various apoptotic stimuli. In nontumorigenic cells, fragments N1 and N2 do not modulate apoptosis. Therefore, the role of the second caspase-mediated cleavage of RasGAP is to allow the inactivation of the antiapoptotic function of fragment N so that caspases are no longer hampered in their ability to kill cells. PMID- 15901832 TI - Cathepsin B regulates the intrinsic angiogenic threshold of endothelial cells. AB - The lysosomal protease cathepsin B has been implicated in a variety of pathologies including pancreatitis, tumor angiogenesis, and neuronal diseases. We used a tube formation assay to investigate the role of cathepsin B in angiogenesis. When cultured between two layers of collagen I, primary endothelial cells formed tubes in response to exogenously added VEGF. Overexpressing cathepsin B reduced the VEGF-dependent tube response, whereas pharmacologically or molecularly suppressing cathepsin B eliminated the dependence on exogenous VEGF. However, tube formation still required VEGF receptor activity, which suggested that endothelial cells generated VEGF. Indeed, VEGF mRNA and protein was detectable in cells treated with cathepsin B inhibitor, which correlated with a rise in the level of HIF-1alpha. In addition to boosting the level of proangiogenic factors, blocking cathepsin B activity reduced the amount of the antiangiogenic protein endostatin. Thus endothelial cells have the intrinsic capacity to generate pro- and antiangiogenic agents. These observations complement and expand our appreciation of how endothelial cell-derived proteases regulate angiogenesis. PMID- 15901833 TI - Basal activation of the P2X7 ATP receptor elevates mitochondrial calcium and potential, increases cellular ATP levels, and promotes serum-independent growth. AB - P2X7 is a bifunctional receptor (P2X7R) for extracellular ATP that, depending on the level of activation, forms a cation-selective channel or a large conductance nonselective pore. The P2X7R has a strong proapoptotic activity but can also support growth. Here, we describe the mechanism involved in growth stimulation. Transfection of P2X7R increases resting mitochondrial potential (delta psi(mt)), basal mitochondrial Ca2+ ([Ca2+]mt), intracellular ATP content, and confers ability to grow in the absence of serum. These changes require a full pore forming function, because they are abolished in cells transfected with a mutated P2X7R that retains channel activity but cannot form the nonselective pore, and depend on an autocrine/paracrine tonic stimulation by secreted ATP. On the other hand, sustained stimulation of P2X7R causes a delta psi(mt) drop, a large increase in [Ca2+]mt, mitochondrial fragmentation, and cell death. These findings reveal a hitherto undescribed mechanism for growth stimulation by a plasma membrane pore. PMID- 15901834 TI - The complex interplay between the neck and hinge domains in kinesin-1 dimerization and motor activity. AB - Kinesin-1 dimerizes via the coiled-coil neck domain. In contrast to animal kinesins, neck dimerization of the fungal kinesin-1 NcKin requires additional residues from the hinge. Using chimeric constructs containing or lacking fungal specific elements, the proximal part of the hinge was shown to stabilize the neck coiled-coil conformation in a complex manner. The conserved fungal kinesin hinge residue W384 caused neck coiled-coil formation in a chimeric NcKin construct, including parts of the human kinesin-1 stalk. The stabilizing effect was retained in a NcKinW384F mutant, suggesting important pi-stacking interactions. Without the stalk, W384 was not sufficient to induce coiled-coil formation, indicating that W384 is part of a cluster of several residues required for neck coiled-coil folding. A W384-less chimera of NcKin and human kinesin possessed a non-coiled coil neck conformation and showed inhibited activity that could be reactivated when artificial interstrand disulfide bonds were used to stabilize the neck coiled-coil conformation. On the basis of yeast two-hybrid data, we propose that the proximal hinge can bind kinesin's cargo-free tail domain and causes inactivation of kinesin by disrupting the neck coiled-coil conformation. PMID- 15901835 TI - Atg17 regulates the magnitude of the autophagic response. AB - Autophagy is a catabolic process used by eukaryotic cells for the degradation and recycling of cytosolic proteins and excess or defective organelles. In yeast, autophagy is primarily a response to nutrient limitation, whereas in higher eukaryotes it also plays a role in developmental processes. Due to its essentially unlimited degradative capacity, it is critical that regulatory mechanisms are in place to modulate the timing and magnitude of the autophagic response. One set of proteins that seems to function in this regard includes a complex that contains the Atg1 kinase. Aside from Atg1, the proteins in this complex participate primarily in either nonspecific autophagy or specific types of autophagy, including the cytoplasm to vacuole targeting pathway, which operates under vegetative growth conditions, and peroxisome degradation. Accordingly, these proteins are prime candidates for factors that regulate the conversion between these pathways, including the change in size of the sequestering vesicle, the most obvious morphological difference. The atg17delta mutant forms a reduced number of small autophagosomes. As a result, it is defective in peroxisome degradation and is partially defective for autophagy. Atg17 interacts with both Atg1 and Atg13, via two coiled-coil domains, and these interactions facilitate its inclusion in the Atg1 complex. PMID- 15901836 TI - Structural requirements for differential sensitivity of KCNQ K+ channels to modulation by Ca2+/calmodulin. AB - Calmodulin modulation of ion channels has emerged as a prominent theme in biology. The sensitivity of KCNQ1-5 K+ channels to modulation by Ca2+/calmodulin (CaM) was studied using patch-clamp, Ca2+ imaging, and biochemical and pharmacological approaches. Coexpression of CaM in Chinese hamster ovary (CHO) cells strongly reduced currents of KCNQ2, KCNQ4, and KCNQ5, but not KCNQ1 or KCNQ3. In simultaneous current recording/Ca2+ imaging experiments, CaM conferred Ca2+ sensitivity to KCNQ4 and KCNQ5, but not to KCNQ1, KCNQ3, or KCNQ1/KCNE1 channels. A chimera constructed from the carboxy terminus of KCNQ4 and the rest KCNQ1 displayed Ca2+ sensitivity similar to KCNQ4. Chimeras constructed from different lengths of the KCNQ4 carboxy terminal and the rest KCNQ3 localized a region that confers sensitivity to Ca2+/CaM. Lobe-specific mutations of CaM revealed that its amino-terminal lobe mediates the Ca2+ sensitivity of the KCNQ/CaM complex. The site of CaM action within the channel carboxy terminus overlaps with that of the KCNQ opener N-ethylmaleimide (NEM). We found that CaM overexpression reduced NEM augmentation of KCNQ2, KCNQ4, and KCNQ5, and NEM pretreatment reduced Ca2+/CaM-mediated suppression of M current in sympathetic neurons by bradykinin. We propose that two functionally distinct types of carboxy termini underlie the observed differences among this channel family. PMID- 15901837 TI - Role of the septin ring in the asymmetric localization of proteins at the mother bud neck in Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, septins form a scaffold in the shape of a ring at the future budding site that rearranges into a collar at the mother-bud neck. Many proteins bind asymmetrically to the septin collar. We found that the protein Bni4-CFP was located on the exterior of the septin ring before budding and on the mother side of the collar after budding, whereas the protein kinase Kcc4-YFP was located on the interior of the septin ring before budding and moved into the bud during the formation of the septin collar. Unbudded cells treated with the actin inhibitor latrunculin-A assembled cortical caps of septins on which Bni4-CFP and Kcc4-YFP colocalized. Bni4-CFP and Kcc4-YFP also colocalized on cortical caps of septins found in strains deleted for the genes encoding the GTPase activating proteins of Cdc42 (RGA1, RGA2, and BEM3). However, Bni4-CFP and Kcc4-YFP were still partially separated in mutants (gin4, elm1, cla4, and cdc3-1) in which septin morphology was severely disrupted in other ways. These observations provide clues to the mechanisms for the asymmetric localization of septin-associated proteins. PMID- 15901838 TI - Unexpected diversity and differential success of DNA transposons in four species of entamoeba protozoans. AB - We report the first comprehensive analysis of transposable element content in the compact genomes (approximately 20 Mb) of four species of Entamoeba unicellular protozoans for which draft sequences are now available. Entamoeba histolytica and Entamoeba dispar, two human parasites, have many retrotransposons, but few DNA transposons. In contrast, the reptile parasite Entamoeba invadens and the free living Entamoeba moshkovskii contain few long interspersed elements but harbor diverse and recently amplified populations of DNA transposons. Representatives of three DNA transposase superfamilies (hobo/Activator/Tam3, Mutator, and piggyBac) were identified for the first time in a protozoan species in addition to a variety of members of a fourth superfamily (Tc1/mariner), previously reported only from ciliates and Trichomonas vaginalis among protozoans. The diversity of DNA transposons and their differential amplification among closely related species with similar compact genomes are discussed in the context of the biology of Entamoeba protozoans. PMID- 15901839 TI - Mitochondrial genome sequences support ancient population expansion in Plasmodium vivax. AB - Examination of nucleotide diversity in 106 mitochondrial genomes of the most geographically widespread human malaria parasite, Plasmodium vivax, revealed a level of diversity similar to, but slightly higher than, that seen in the virulent human malaria parasite Plasmodium falciparum. The pairwise distribution of nucleotide differences among mitochondrial genome sequences supported the hypothesis that both these parasites underwent ancient population expansions. We estimated the age of the most recent common ancestor (MRCA) of the mitochondrial genomes of both P. vivax and P. falciparum at around 200,000-300,000 years ago. This is close to the previous estimates of the time of the human mitochondrial MRCA and the origin of modern Homo sapiens, consistent with the hypothesis that both these Plasmodium species were parasites of the hominid lineage before the origin of modern H. sapiens and that their population expansion coincided with the population expansion of their host. PMID- 15901840 TI - Mathematical modeling of evolution of horizontally transferred genes. AB - We describe a stochastic birth-and-death model of evolution of horizontally transferred genes in microbial populations. The model is a generalization of the stochastic model described by Berg and Kurland and includes five parameters: the rate of mutational inactivation, selection coefficient, invasion rate (i.e., rate of arrival of a novel sequence from outside of the recipient population), within population horizontal transmission ("infection") rate, and population size. The model of Berg and Kurland included four parameters, namely, mutational inactivation, selection coefficient, population size, and "infection." However, the effect of "infection" was disregarded in the interpretation of the results, and the overall conclusion was that horizontally acquired sequences can be fixed in a population only when they confer a substantial selective advantage onto the recipient and therefore are subject to strong positive selection. Analysis of the present model in different domains of parameter values shows that, as long as the rate of within-population horizontal transmission is comparable to the mutational inactivation rate and there is even a low rate of invasion, horizontally acquired sequences can be fixed in the population or at least persist for a long time in a substantial fraction of individuals in the population even when they are neutral or slightly deleterious. The available biological data strongly suggest that intense within-population and even between-populations gene flows are realistic for at least some prokaryotic species and environments. Therefore, our modeling results are compatible with the notion of a pivotal role of horizontal gene transfer in the evolution of prokaryotes. PMID- 15901841 TI - Interchromosomal segmental duplications explain the unusual structure of PRSS3, the gene for an inhibitor-resistant trypsinogen. AB - Homo sapiens possess several trypsinogen or trypsinogen-like genes of which three (PRSS1, PRSS2, and PRSS3) produce functional trypsins in the digestive tract. PRSS1 and PRSS2 are located on chromosome 7q35, while PRSS3 is found on chromosome 9p13. Here, we report a variation of the theme of new gene creation by duplication: the PRSS3 gene was formed by segmental duplications originating from chromosomes 7q35 and 11q24. As a result, PRSS3 transcripts display two variants of exon 1. The PRSS3 transcript whose gene organization most resembles PRSS1 and PRSS2 encodes a functional protein originally named mesotrypsinogen. The other variant is a fusion transcript, called trypsinogen IV. We show that the first exon of trypsinogen IV is derived from the noncoding first exon of LOC120224, a chromosome 11 gene. LOC120224 codes for a widely conserved transmembrane protein of unknown function. Comparative analyses suggest that these interchromosomal duplications occurred after the divergence of Old World monkeys and hominids. PRSS3 transcripts consist of a mixed population of mRNAs, some expressed in the pancreas and encoding an apparently functional trypsinogen and others of unknown function expressed in brain and a variety of other tissues. Analysis of the selection pressures acting on the trypsinogen gene family shows that, while the apparently functional genes are under mild to strong purifying selection overall, a few residues appear under positive selection. These residues could be involved in interactions with inhibitors. PMID- 15901842 TI - Comparative phylogenetic analysis of blcap/nnat reveals eutherian-specific imprinted gene. AB - Imprinted genes are parent-of-origin dependent, monoallelically expressed genes present in marsupials and eutherian mammals. Altered expression of imprinted genes plays a significant role in the etiology of a variety of human disorders and diseases. Nevertheless, the regulatory mechanisms of imprinting remain poorly defined. The imprinted gene Neuronatin (Nnat) is an excellent candidate for studying imprinting because it resides within the 8.5-kb intron of the nonimprinted gene Bladder Cancer-Associated Protein (Blcap) and is the only imprinted gene within the region. A phylogenetic comparison of this micro imprinted domain in human, mouse, and rat revealed several candidates for imprint control, including tandem repeats and putative binding sites for trans- acting factors known to be involved in chromatin remodeling. Genome-wide phylogenetic comparisons of species from the three major extant mammalian clades failed, however, to show any evidence of Nnat outside the eutherian lineage. Thus, Nnat is the first identified eutherian-specific imprinted gene, demonstrating that imprinted genes did not arise at a single point during evolution. This finding also suggests that the complexity of imprinting regulation observed at other loci may, in part, be directly related to the amount of time they have been imprinted. PMID- 15901843 TI - Alu-SINE exonization: en route to protein-coding function. AB - The majority of more than one million primate-specific Alu elements map to nonfunctional parts of introns or intergenic sequences. Once integrated, they have the potential to become exapted as functional modules, e.g., as protein coding domains via alternative splicing. This particular process is also termed exonization and increases protein versatility. Here we investigate 153 human chromosomal loci where Alu elements were conceivably exonized. In four selected examples, we generated, with the aid of representatives of all primate infraorders, phylogenetic reconstructions of the evolutionary steps presumably leading to exonization of Alu elements. We observed a variety of possible scenarios in which Alu elements led to novel mRNA splice forms and which, like most evolutionary processes, took different courses in different lineages. Our data show that, once acquired, some exonizations were lost again in some lineages. In general, Alu exonization occurred at various time points over the evolutionary history of primate lineages, and protein-coding potential was acquired either relatively soon after integration or millions of years thereafter. The course of these paths can probably be generalized to the exonization of other elements as well. PMID- 15901844 TI - Expression of adamalysin 19/ADAM19 in the endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy. AB - A disintegrin and metalloproteinase (ADAM) 19 may contribute to multiple processes including proteolysis, adhesion and intracellular signalling. These processes are also critical for embryo implantation. The aim of this study was to investigate the spatio-temporal expression of the ADAM19 in rhesus monkey uteri on days 12, 18 and 26 of pregnancy. The results showed that in the cloned monkey 346 bp ADAM19 gene fragment and 114 amino acid residues were 98 and 100% identical to those of human homologues, respectively. In-situ hybridization confirmed that the ADAM19 mRNA was located in the luminal and glandular epithelium on day 12 of pregnancy. On day 18 of pregnancy, strong signals of the ADAM19 mRNA were detected in the placental villi, trophoblastic column and glandular epithelium near the myometrium. Moderate expression of the ADAM19 mRNA was seen in the trophoblastic shell and stromal cells. The placental villi and trophoblastic column expressed abundant ADAM19 mRNA, and ADAM19 transcripts were also detected in the trophoblastic shell and fetal-maternal border on day 26 of pregnancy. The expression pattern of the ADAM19 protein was similar to its transcript, but signals for the ADAM19 protein in the stromal cells and trophoblastic shell increased more than its mRNA on day 18 of pregnancy. Statistical analysis demonstrated that the expression level of ADAM19 significantly increased on day 18 of pregnancy. These data suggest that the ADAM19 may be involved in the key processes of glandular secretion, trophoblast invasion and degradation of extracellular matrix during early pregnancy. PMID- 15901845 TI - Tumour necrosis factor-alpha gene haplotype is associated with pre-eclampsia. AB - We determined whether polymorphisms in the promoter region of the tumour necrosis factor alpha (TNF-alpha) gene contributes to differences in susceptibility to develop pre-eclampsia. The study involved 133 pre-eclamptic and 115 healthy pregnant women who were genotyped for the G-308A polymorphism of the TNF-alpha gene. The frequency of the G-308A allele was more common in the pre-eclampsia group than among the controls (P=0.046), giving an odds ratio of 0.57 (95% CI: 0.32-0.99), but there were no differences in the genotype distribution. The data from the G-308A polymorphism was combined with the previously published genotype and allele data from the C-850T polymorphism of the TNF-alpha gene, and used to assess a haplotype estimation analysis. Estimated overall pair of loci haplotype frequencies differed significantly between the groups (P=0.023+/-0.004). In the single haplotype association analysis, the haplotype C-A versus others was over represented in the pre-eclampsia group (P=0.041+/-0.003), whereas the haplotype T G versus others was less common in the pre-eclampsia group (P=0.035+/-0.003), compared with the controls. In conclusion, the polymorphisms of the TNF-alpha gene showed a significant haplotype association with susceptibility to pre eclampsia in the Finnish population. PMID- 15901846 TI - Long-term morphine treatment enhances proteasome-dependent degradation of G beta in human neuroblastoma SH-SY5Y cells: correlation with onset of adenylate cyclase sensitization. AB - The initial aim of this study was to identify protein changes associated with long-term morphine treatment in a recombinant human neuroblastoma SH-SY5Y clone (sc2) stably overexpressing the human mu-opioid (MOP) receptor. In MOP receptor overexpressing sc2 cells, short-term morphine exposure was found to be much more potent and efficacious in inhibiting forskolin-elicited production of cAMP, and long-term morphine exposure was shown to induce a substantially higher degree of opiate dependence, as reflected by adenylate cyclase sensitization, than it did in wild-type neuroblastoma cells. Differential proteomic analysis of detergent resistant membrane rafts isolated from untreated and chronically morphine-treated sc2 cells revealed long-term morphine exposure to have reliably induced a 30 to 40% decrease in the abundance of five proteins, subsequently identified by mass spectrometry as G protein subunits alphai(2), alphai(3), beta(1), and beta(2), and prohibitin. Quantitative Western blot analyses of whole-cell extracts showed that long-term morphine treatment-induced down-regulation of Gbeta but not of the other proteins is highly correlated (r(2) = 0.96) with sensitization of adenylate cyclase. Down-regulation of Gbeta and adenylate cyclase sensitization elicited by long-term morphine treatment were suppressed in the presence of carbobenzoxy-l leucyl-l-leucyl-l-norvalinal (MG-115) or lactacystin. Thus, sustained activation of the MOP receptor by morphine in sc2 cells seems to promote proteasomal degradation of Gbeta to sensitize adenylate cyclase. Together, our data suggest that the long-term administration of opiates may elicit dependence by altering the neuronal balance of heterotrimeric G proteins and adenylate cyclases, with the ubiquitin-proteasome pathway playing a pivotal role. PMID- 15901847 TI - Diketo hexenoic acid derivatives are novel selective non-nucleoside inhibitors of mammalian terminal deoxynucleotidyl transferases, with potent cytotoxic effect against leukemic cells. AB - Mammalian terminal deoxyribonucleotidyl transferase (TDT) catalyzes the non template-directed polymerization of deoxyribonucleoside triphosphates and has a key role in V(D)J recombination during lymphocyte and repertoire development. More than 90% of leukemic cells in acute lymphocytic leukemia and approximately 30% of leukemic cells in the chronic myelogenous leukemia crisis show elevated TDT activity. This finding is connected to a poor prognosis and response to chemotherapy and reduced survival time. On the other hand, recent data indicated that TDT is not the only terminal deoxyribonucleotidyl transferase in mammalian cells. Its close relative, DNA polymerase lambda, can synthesize DNA both in a template-dependent (polymerase) and template-independent (terminal deoxyribonucleotidyl transferase) fashion. DNA polymerase lambda might be involved in the nonhomologous end-joining recombinational repair pathway of DNA double-strand breaks. In this work, we report the characterization of the mechanism of action of three diketo hexenoic acid (DKHA) derivatives, which proved to be extremely selective for the terminal deoxyribonucleotidyl transferase activity of DNA polymerase lambda and TDT. They seem to be the first non-nucleoside-specific inhibitors of mammalian terminal transferases reported. Moreover, the DKHA analog 6-(1-phenylmethyl-1H-indol-3-yl)-2,4-dioxo-5-hexenoic acid (RDS2119) was not toxic toward HeLa cells (CC(50) > 100 muM), whereas it showed significant cytotoxicity against the TDT(+) leukemia cell line MOLT-4 (CC(50) = 14.9 muM), thus having the potential to be further developed as a novel antitumor agent. PMID- 15901848 TI - Single mutations at Asn295 and Leu305 in the cytoplasmic half of transmembrane alpha-helix domain 7 of the AT1 receptor induce promiscuous agonist specificity for angiotensin II fragments: a pseudo-constitutive activity. AB - The most striking feature of a G protein-coupled receptor (GPCR) is its highly exclusive agonist specificity. This feature guarantees that a GPCR recognizes only its specific native agonist(s). In this study, we showed that two point mutations of N295S and L305Q enabled the AT(1) receptors to recognize multiple Ang II fragments. Similar to the well established constitutively active AT(1) mutant receptor N111G, the mutations of N295S and L305Q induced an increased production of basal inositol 1,4,5-phosphates in the absence of exogenous Ang II when expressed in HEK293 cells. Distinct from the N111G, however, is the fact that the increased basal activity disappeared in COS-7 cells because of the lack of endogenous Ang II fragments produced by the cells-a pseudo-constitutive activity. It is surprising that the Ang II analog [Sar(1),Ile(4),Ile(8)]Ang II and the native angiotensin II fragments Ang 1-7, Ang IV, and Ang 5-8, which are inactive in activating the wild-type receptor, activated N295S and L305Q. Results generated by lowering the Na(+) concentration suggest that the mutant N295S and L305Q may be trapped in neutral conformational states (R(N)). These data allow us to identify for the first time a novel pattern of GPCR mutations with a broad spectrum of agonist specificity, suggesting possible existence of functional GPCRs in nature that are activated through conformational "selection" rather than "induction" mechanisms. PMID- 15901849 TI - Mutations linked to autosomal dominant nocturnal frontal lobe epilepsy affect allosteric Ca2+ activation of the alpha 4 beta 2 nicotinic acetylcholine receptor. AB - Extracellular Ca(2+) robustly potentiates the acetylcholine response of alpha4beta2 nicotinic receptors. Rat orthologs of five mutations linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE)-alpha4(S252F), alpha4(S256L), alpha4(+L264), beta2(V262L), and beta2(V262M)-reduced 2 mM Ca(2+) potentiation of the alpha4beta2 1 mM acetylcholine response by 55 to 74%. To determine whether altered allosteric Ca(2+) activation or enhanced Ca(2+) block caused this reduction, we coexpressed the rat ADNFLE mutations with an alpha4 N terminal mutation, alpha4(E180Q), that abolished alpha4beta2 allosteric Ca(2+) activation. In each case, Ca(2+) inhibition of the double mutants was less than that expected from a Ca(2+) blocking mechanism. In fact, the effects of Ca(2+) on the ADNFLE mutations near the intracellular end of the M2 region-alpha4(S252F) and alpha4(S256L)-were consistent with a straightforward allosteric mechanism. In contrast, the effects of Ca(2+) on the ADNFLE mutations near the extracellular end of the M2 region-alpha4(+L264)beta2, beta2(V262L), and beta2(V262M)-were consistent with a mixed mechanism involving both altered allosteric activation and enhanced block. However, the effects of 2 mM Ca(2+) on the alpha4beta2, alpha4(+L264)beta2, and alpha4beta2(V262L) single-channel conductances, the effects of membrane potential on the beta2(V262L)-mediated reduction in Ca(2+) potentiation, and the effects of eliminating the negative charges in the extracellular ring on this reduction failed to provide any direct evidence of mutant-enhanced Ca(2+) block. Moreover, analyses of the alpha4beta2, alpha4(S256L), and alpha4(+L264) Ca(2+) concentration-potentiation relations suggested that the ADNFLE mutations reduce Ca(2+) potentiation of the alpha4beta2 acetylcholine response by altering allosteric activation rather than by enhancing block. PMID- 15901850 TI - Expression profiling of ABC transporters in a drug-resistant breast cancer cell line using AmpArray. AB - ATP-binding cassette (ABC) membrane proteins comprise a superfamily of transporters with a wide variety of substrates. Humans have 49 members in this superfamily. Several human ABC transporters, such as ABCB1 and ABCC1, have been attributed to cause multidrug resistance (MDR) in cancer treatment when over expressed. In the past, an MDR cancer cell line MCF7/AdVp3000 has been selected, and overexpression of ABCG2 was thought to cause MDR in this cell line. However, ectopic overexpression of ABCG2 in MCF7 cells could not explain the high drug resistance level observed with the selected cell line. In this study, we designed an AmpArray analysis to profile whether other ABC transporters were also selected to contribute to the increased drug resistance in MCF7/AdVp3000 cells. We found that 16 ABC transporters, including ABCG2, had >/=1.5-fold altered expression in MCF7/AdVp3000 compared with the parental MCF7 cells. In particular, the expression of ABCA4 and ABCC3 was increased by 132- and 459-fold, respectively, whereas ABCG2 was increased by approximately 3000-fold. Furthermore, the elevated expression of these three transporters reversed with the reversed drug resistance phenotype, and silencing ABCC3 expression in MCF7/AdVp3000 cells significantly reduced doxorubicin resistance. Thus, other ABC transporters in addition to ABCG2 are likely to contribute to the MDR selected in MCF7/AdVp3000 cells. This study also shows that AmpArray can be used as a quick and easy tool to profile the expression of ABC transporters in resistant cell lines and tumor samples for potential use in individualized design of therapy. PMID- 15901851 TI - Integrin engagement increases histone H3 acetylation and reduces histone H1 association with DNA in murine lung endothelial cells. AB - Engagement of integrin cell adhesion receptors in mouse lung endothelial cells induces global sensitivity of DNA to nuclease digestion, reflecting alterations in chromatin structure. These structural changes may contribute to the antigenotoxic effects of integrin engagement in lung endothelium. Because histone acetylation and poly(ADP-ribosyl)ation modulate chromatin structure, we investigated the effects of beta1 integrin engagement with antibody on these post translational modifications and the presence of histones at discrete DNA sequences in the mouse lung endothelial cell genome using chromatin immunoprecipitation. Integrin engagement increased acetylation of core histone H3. The presence of acetylated histone H3 at intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) promoters, and a nonpromoter sequence was also increased. As with integrin engagement, the histone deacetylase inhibitor trichostatin A caused global hypersensitivity of DNA to nuclease digestion and induced acetylation of histone H3 and its coimmunoprecipitation with VCAM-1 and ICAM-1 promoters and nonpromoter DNA. In contrast to acetyl-histone H3, the association of linker histone H1 with specific DNA sequences was either reduced or unaffected by integrin engagement and trichostatin A. Although integrin engagement and trichostatin A treatment did not affect histone H1 poly(ADP-ribosyl)ation, deletion of poly(ADP-ribose) polymerase 1 increased core histone H3 acetylation and increased its level at the iNOS promoter while decreasing the amount of histone H1. The results suggest that integrin engagement, as well as trichostatin A and PARP-1 deletion, regulate chromatin structure via core histone H3 acetylation and reduced linker histone H1 DNA association. PMID- 15901852 TI - Characterization of a human carcinoma cell line selected for resistance to the farnesyl transferase inhibitor 4-(2-(4-(8-chloro-3,10-dibromo-6,11-dihydro-5H benzo-(5,6)-cyclohepta(1,2-b)-pyridin-11(R)-yl)-1-piperidinyl)-2-oxo-ethyl)-1 piperidinecarboxamide (SCH66336). AB - Farnesyl protein transferase inhibitors (FTIs) have demonstrated clinical activity in certain solid tumors and hematological malignancies. Little is known about mechanisms of resistance to these agents. To provide a basis for better understanding FTI resistance, the colorectal carcinoma cell line HCT 116 was selected by stepwise exposure to increasing 4-(2-(4-(8-chloro-3,10-dibromo-6,11 dihydro-5H-benzo-(5,6)-cyclohepta(1,2-b)-pyridin-11(R)-yl)-1-piperidinyl)-2-oxo ethyl)-1-piperidinecarboxamide (SCH66336) concentrations. The resulting line, HCT 116R, was 100-fold resistant to SCH66336 and other FTIs, including methyl {N-[2 phenyl-4-N[2(R)-amino-3-mercaptopropylamino] benzoyl]}-methionate (FTI-277), but was less than 2-fold resistant to the standard agents gemcitabine, cisplatin, and paclitaxel. Accumulation of the unfarnesylated forms of prelamin A and HDJ-2, two substrates that reflect farnesyl transferase inhibition, was similar in FTI treated parental and HCT 116R cells, indicating that alterations in drug uptake or inhibition of farnesyl protein transferase is not the mechanism of resistance. Changes in signal-transduction pathways that might account for this resistance were examined by immunoblotting and confirmed pharmacologically. There was no difference in activation of the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase pathway or sensitivity to the MEK1/2 inhibitor 2'-amino-3'-methoxyflavone (PD98059) in HCT 116R cells. In contrast, increased phosphorylation of the molecular target of rapamycin (mTOR) and its downstream target p70 S6 kinase and increased levels of Akt1 and Akt2 were demonstrated in HCT 116R cells. Further experiments demonstrated that the mTOR inhibitor rapamycin selectively sensitized HCT 116R cells to SCH66336 but not to gemcitabine, cisplatin, or paclitaxel. These findings provide evidence that alterations in the phosphatidylinositol-3 kinase/Akt pathway can contribute to FTI resistance and suggest a potential strategy for overcoming this resistance. PMID- 15901853 TI - Offset recombinant PCR: a simple but effective method for shuffling compact heterologous domains. AB - DNA shuffling and other in vitro recombination strategies have proven highly effective at generating complex libraries for mutagenesis studies. While most recombination techniques employ DNA polymerases in part of a multi-step process, few seek to exploit the natural recombinogenic tendencies and exponential amplification rates of PCR. Here, we characterize a simple but effective method for using standard PCR to promote high recombination frequencies among compact heterologous domains by locating the domains near one end of the template. In a typical amplification reaction, Pfu polymerase generated chimeric crossover events in 13% of the population when markers were separated by only 70 nt. The fraction of recombinant sequences reached 42% after six consecutive rounds of PCR, a value close to the 50% expected from a fully shuffled population. When homology within the recombinant region was reduced to 82%, the recombination frequency dropped by nearly half for a single amplification reaction and crossover events were clustered toward one end of the domain. Surprisingly, recombination frequencies for template populations with high and low sequence homologies converged after just four rounds of PCR, suggesting that the exponential accumulation of chimeric molecules in the PCR mixture serves to promote recombination within heterologous domains. PMID- 15901854 TI - Prediction of functional modules based on comparative genome analysis and Gene Ontology application. AB - We present a computational method for the prediction of functional modules encoded in microbial genomes. In this work, we have also developed a formal measure to quantify the degree of consistency between the predicted and the known modules, and have carried out statistical significance analysis of consistency measures. We first evaluate the functional relationship between two genes from three different perspectives--phylogenetic profile analysis, gene neighborhood analysis and Gene Ontology assignments. We then combine the three different sources of information in the framework of Bayesian inference, and we use the combined information to measure the strength of gene functional relationship. Finally, we apply a threshold-based method to predict functional modules. By applying this method to Escherichia coli K12, we have predicted 185 functional modules. Our predictions are highly consistent with the previously known functional modules in E.coli. The application results have demonstrated that our approach is highly promising for the prediction of functional modules encoded in a microbial genome. PMID- 15901855 TI - Images in clinical medicine. Chromoblastomycosis. PMID- 15901856 TI - Traumatic brain injury in the war zone. PMID- 15901857 TI - Perinatal mortality in developing countries. PMID- 15901858 TI - Cystatin C and the risk of death and cardiovascular events among elderly persons. AB - BACKGROUND: Cystatin C is a serum measure of renal function that appears to be independent of age, sex, and lean muscle mass. We compared creatinine and cystatin C levels as predictors of mortality from cardiovascular causes and from all causes in the Cardiovascular Health Study, a cohort study of elderly persons living in the community. METHODS: Creatinine and cystatin C were measured in serum samples collected from 4637 participants at the study visit in 1992 or 1993; follow-up continued until June 30, 2001. For each measure, the study population was divided into quintiles, with the fifth quintile subdivided into thirds (designated 5a, 5b, and 5c). RESULTS: Higher cystatin C levels were directly associated, in a dose-response manner, with a higher risk of death from all causes. As compared with the first quintile, the hazard ratios (and 95 percent confidence intervals) for death were as follows: second quintile, 1.08 (0.86 to 1.35); third quintile, 1.23 (1.00 to 1.53); fourth quintile, 1.34 (1.09 to 1.66); quintile 5a, 1.77 (1.34 to 2.26); 5b, 2.18 (1.72 to 2.78); and 5c, 2.58 (2.03 to 3.27). In contrast, the association of creatinine categories with mortality from all causes appeared to be J-shaped. As compared with the two lowest quintiles combined (cystatin C level, < or =0.99 mg per liter), the highest quintile of cystatin C (> or =1.29 mg per liter) was associated with a significantly elevated risk of death from cardiovascular causes (hazard ratio, 2.27 [1.73 to 2.97]), myocardial infarction (hazard ratio, 1.48 [1.08 to 2.02]), and stroke (hazard ratio, 1.47 [ 1.09 to 1.96]) after multivariate adjustment. The fifth quintile of creatinine, as compared with the first quintile, was not independently associated with any of these three outcomes. CONCLUSIONS: Cystatin C, a serum measure of renal function, is a stronger predictor of the risk of death and cardiovascular events in elderly persons than is creatinine. PMID- 15901859 TI - Colonoscopic screening of average-risk women for colorectal neoplasia. AB - BACKGROUND: Veterans Affairs (VA) Cooperative Study 380 showed that some advanced colorectal neoplasias (i.e., adenomas at least 1 cm in diameter, villous adenomas, adenomas with high-grade dysplasia, or cancer) in men would be missed with the use of flexible sigmoidoscopy but detected by colonoscopy. In a tandem study, we examined the yield of screening colonoscopy in women. METHODS: To determine the prevalence and location of advanced neoplasia, we offered colonoscopy to consecutive asymptomatic women referred for colon-cancer screening. The diagnostic yield of flexible sigmoidoscopy was calculated by estimating the proportion of patients with advanced neoplasia whose lesions would have been identified if they had undergone flexible sigmoidoscopy alone. Lesions were considered detectable by flexible sigmoidoscopy if they were in the distal colon or if they were in the proximal colon in patients who had concurrent small adenomas in the distal colon, a finding that would have led to colonoscopy. The results were compared with the results from VA Cooperative Study 380 for age matched men and women with negative fecal occult-blood tests and no family history of colon cancer. RESULTS: Colonoscopy was complete in 1463 women, 230 of whom (15.7 percent) had a family history of colon cancer. Colonoscopy revealed advanced neoplasia in 72 women (4.9 percent). If flexible sigmoidoscopy alone had been performed, advanced neoplasia would have been detected in 1.7 percent of these women (25 of 1463) and missed in 3.2 percent (47 of 1463). Only 35.2 percent of women with advanced neoplasia would have had their lesions identified if they had undergone flexible sigmoidoscopy alone, as compared with 66.3 percent of matched men from VA Cooperative Study 380 (P<0.001). CONCLUSIONS: Colonoscopy may be the preferred method of screening for colorectal cancer in women. PMID- 15901860 TI - Transplantation of umbilical-cord blood in babies with infantile Krabbe's disease. AB - BACKGROUND: Infantile Krabbe's disease produces progressive neurologic deterioration and death in early childhood. We hypothesized that transplantation of umbilical-cord blood from unrelated donors before the development of symptoms would favorably alter the natural history of the disease among newborns in whom the disease was diagnosed because of a family history. We compared the outcomes among these newborns with the outcomes among infants who underwent transplantation after the development of symptoms and with the outcomes in an untreated cohort of affected children. METHODS: Eleven asymptomatic newborns (age range, 12 to 44 days) and 14 symptomatic infants (age range, 142 to 352 days) with infantile Krabbe's disease underwent transplantation of umbilical-cord blood from unrelated donors after myeloablative chemotherapy. Engraftment, survival, and neurodevelopmental function were evaluated longitudinally for four months to six years. RESULTS: The rates of donor-cell engraftment and survival were 100 percent and 100 percent, respectively, among the asymptomatic newborns (median follow-up, 3.0 years) and 100 percent and 43 percent, respectively, among the symptomatic infants (median follow-up, 3.4 years). Surviving patients showed durable engraftment of donor-derived hematopoietic cells with restoration of normal blood galactocerebrosidase levels. Infants who underwent transplantation before the development of symptoms showed progressive central myelination and continued gains in developmental skills, and most had age-appropriate cognitive function and receptive language skills, but a few had mild-to-moderate delays in expressive language and mild-to-severe delays in gross motor function. Children who underwent transplantation after the onset of symptoms had minimal neurologic improvement. CONCLUSIONS: Transplantation of umbilical-cord blood from unrelated donors in newborns with infantile Krabbe's disease favorably altered the natural history of the disease. Transplantation in babies after symptoms had developed did not result in substantive neurologic improvement. PMID- 15901861 TI - Asthma as a risk factor for invasive pneumococcal disease. AB - BACKGROUND: The risk of invasive pneumococcal disease among persons with asthma is unknown. METHODS: We conducted a nested case-control study to examine the association between asthma and invasive pneumococcal disease. The study population included persons 2 to 49 years of age who were enrolled in Tennessee's Medicaid program (TennCare) for more than one year during the study period (1995 through 2002) and who resided in counties participating in a prospective laboratory-based program of surveillance for invasive pneumococcal disease. For each subject with invasive pneumococcal disease, 10 age-matched controls without invasive pneumococcal disease were randomly selected from the same population. TennCare files were queried to identify the presence of coexisting conditions that confer a high risk of pneumococcal disease. For the purpose of our study, asthma was defined by documentation of one or more inpatient or emergency department diagnoses of asthma, two outpatient diagnoses, or the use of asthma related medications. High-risk asthma was defined as asthma requiring admission to a hospital or a visit to an emergency department, the use of rescue therapy or long-term use of oral corticosteroids, or the dispensing of three or more prescriptions for beta-agonists within the year before enrollment in the study. RESULTS: A total of 635 persons with invasive pneumococcal disease and 6350 controls were identified, of whom 114 (18.0 percent) and 516 (8.1 percent), respectively, had asthma. Persons with asthma had an increased risk of invasive pneumococcal disease (adjusted odds ratio, 2.4; 95 percent confidence interval, 1.9 to 3.1) as compared with controls. Among those without coexisting conditions, the annual incidence of invasive pneumococcal disease was 4.2 episodes per 10,000 persons with high-risk asthma and 2.3 episodes per 10,000 persons with low-risk asthma, as compared with 1.2 episodes per 10,000 persons without asthma. CONCLUSIONS: Asthma is an independent risk factor for invasive pneumococcal disease. The risk among persons with asthma was at least double that among controls. PMID- 15901862 TI - An intervention involving traditional birth attendants and perinatal and maternal mortality in Pakistan. AB - BACKGROUND: There are approximately 4 million neonatal deaths and half a million maternal deaths worldwide each year. There is limited evidence from clinical trials to guide the development of effective maternity services in developing countries. METHODS: We performed a cluster-randomized, controlled trial involving seven subdistricts (talukas) of a rural district in Pakistan. In three talukas randomly assigned to the intervention group, traditional birth attendants were trained and issued disposable delivery kits; Lady Health Workers linked traditional birth attendants with established services and documented processes and outcomes; and obstetrical teams provided outreach clinics for antenatal care. Women in the four control talukas received usual care. The primary outcome measures were perinatal and maternal mortality. RESULTS: Of the estimated number of eligible women in the seven talukas, 10,114 (84.3 percent) were recruited in the three intervention talukas, and 9443 (78.7 percent) in the four control talukas. In the intervention group, 9184 women (90.8 percent) received antenatal care by trained traditional birth attendants, 1634 women (16.2 percent) were seen antenatally at least once by the obstetrical teams, and 8172 safe-delivery kits were used. As compared with the control talukas, the intervention talukas had a cluster-adjusted odds ratio for perinatal death of 0.70 (95 percent confidence interval, 0.59 to 0.82) and for maternal mortality of 0.74 (95 percent confidence interval, 0.45 to 1.23). CONCLUSIONS: Training traditional birth attendants and integrating them into an improved health care system were achievable and effective in reducing perinatal mortality. This model could result in large improvements in perinatal and maternal health in developing countries. PMID- 15901863 TI - Clinical practice. Overweight children and adolescents. PMID- 15901864 TI - Images in clinical medicine. Asphyxia due to an inhaled foreign body. PMID- 15901865 TI - Case records of the Massachusetts General Hospital. Case 15-2005. An 80-year-old man with shortness of breath, edema, and proteinuria. PMID- 15901866 TI - Using every resource to care for our casualties. PMID- 15901867 TI - Chronic kidney disease in the elderly--how to assess risk. PMID- 15901868 TI - Early use of drastic therapy. PMID- 15901869 TI - Unspeakably cruel--torture, medical ethics, and the law. PMID- 15901870 TI - Recombinant activated factor VII for acute intracerebral hemorrhage. PMID- 15901871 TI - Treatment of myelodysplastic syndromes. PMID- 15901872 TI - EGFR mutation and response of lung cancer to gefitinib. PMID- 15901873 TI - Cost-effectiveness of screening for HIV. PMID- 15901874 TI - Unhealthy alcohol use. PMID- 15901875 TI - The unturned stone. PMID- 15901876 TI - Zidovudine and red-cell distribution width. PMID- 15901878 TI - Out of Africa. PMID- 15901879 TI - Mercury from dental amalgam: looking beyond the average. PMID- 15901880 TI - The London Underground: time for a thorough clean-up? PMID- 15901881 TI - The London Underground: dust and hazards to health. AB - AIMS: To assess hazards associated with exposure to dust in the London Underground railway and to provide an informed opinion on the risks to workers and the travelling public of exposure to tunnel dust. METHODS: Concentrations of dust, as mass (PM2.5) and particle number, were measured at different underground stations and in train cabs; its size and composition were analysed; likely maximal exposures of staff and passengers were estimated; and in vitro toxicological testing of sample dusts in comparison with other dusts was performed. RESULTS: Concentrations on station platforms were 270-480 microg/m3 PM2.5 and 14,000-29,000 particles/cm3. Cab concentrations over a shift averaged 130-200 microg/m3 and 17,000-23,000 particles/cm3. The dust comprised by mass approximately 67% iron oxide, 1-2% quartz, and traces of other metals, the residue being volatile matter. The finest particles are drawn underground from the surface while the coarser dust is generated by interaction of brakes, wheels, and rails. Taking account of durations of exposure, drivers and station staff would have maximum exposures of about 200 microg/m3 over eight hours; the occupational exposure standard for welding fume, as iron oxide, is 5 mg/m3 over an eight hour shift. Toxicology showed the dust to have cytotoxic and inflammatory potential at high doses, consistent with its composition largely of iron oxide. DISCUSSION: It is unjustifiable to compare PM2.5 exposure underground with that on the surface, since the adverse effects of iron oxide and combustion generated particles differ. Concentrations of ultrafine particles are lower and of coarser (PM2.5) particles higher underground than on the surface. The concentrations underground are well below allowable workplace concentrations for iron oxide and unlikely to represent a significant cumulative risk to the health of workers or commuters. PMID- 15901882 TI - Serum markers of collagen metabolism: construction workers compared to sedentary workers. AB - BACKGROUND: Evaluation of causal relations between physical load and musculoskeletal disorders is hampered by the lack of knowledge as to the biological relevance of different loading parameters and the large variability between individuals. As indicators of molecular changes in the extracellular matrices of structures of the musculoskeletal system, biomarkers of collagen metabolism may provide important information on biological effects of physical load. The carboxyterminal propeptide of type I collagen (PICP) is a serum marker of synthesis and the carboxyterminal telopeptide region of type I collagen (CTx) reflects degradation of type I collagen. AIMS: To explore the feasibility of biomarkers of type I collagen metabolism as measures of the effects of physical load at tissue level. METHODS: Serum concentrations of PICP and CTx were assessed in a group of male construction workers involved in heavy manual materials handling (n = 47) and in a group of male sedentary workers (n = 49). RESULTS: Serum concentrations of both PICP and CTx seemed to be related to heavy physical work. The ratio PICP/CTx, illustrative of the effective metabolic changes, did not differ between the two groups. CONCLUSIONS: The higher turnover rate but similar effective synthesis may be indicative of an increased type I collagen content in the connective tissues as a result of adaptive remodelling in response to years of exposure to physical load. Further validation of these biomarkers is required with respect to dose-response relations and temporal associations between exposure to back load and biomarker concentrations. PMID- 15901883 TI - Urinary mercury concentrations associated with dental restorations in adult women aged 16-49 years: United States, 1999-2000. AB - BACKGROUND: Mercury amalgam dental restorations have been used by dentists since the mid 19th century and issues on safety continue to be periodically debated within the scientific and public health communities. Previous studies have reported a positive association between urine mercury levels and the number of dental amalgams, but this relation has never been described in a nationally representative sample in the United States. AIMS AND METHODS: Using household interview, dietary interview, dental examination, and laboratory data from the 1999-2000 National Health and Nutrition Examination Survey (NHANES), the association between mercury concentrations and dental restorations was examined in US women of reproductive age. RESULTS: In women of childbearing age, approximately 13% of all posterior dental surfaces were restored with amalgams and the average urinary mercury level in women was low (1.34 microg/l). It is estimated that an increase of 1.8 microg/l in the log transformed values for mercury in urine would occur for each 10 dental surfaces restored with amalgam. CONCLUSIONS: Although the findings do not address the important issues of adverse health effects at low thresholds of mercury exposure, they do provide important reference data that should contribute significantly to the ongoing scientific and public health policy debate on the use of dental amalgams in the USA. PMID- 15901884 TI - Proximity of the home to roads and the risk of wheeze in an Ethiopian population. AB - BACKGROUND: There is widespread public concern that exposure to road vehicle traffic pollution causes asthma, but epidemiological studies in developed countries have not generally confirmed a strong effect and may have underestimated the risk as a result of relatively high and widespread exposure to traffic in everyday life. AIMS: To investigate the effect of living close to a traffic bearing road on the risk of wheezing in Jimma, Ethiopia where road traffic is generally low and restricted to a limited network of roads. METHODS: Data have been previously collected on respiratory symptoms, allergic sensitisation, and numerous demographic and lifestyle factors in a systematic sample of inhabitants of Jimma town. In 2003 the homes of these people were retraced; the shortest distance to the nearest surfaced road, and traffic flows on these roads were measured. RESULTS: Distance measurements were collected for 7609 (80%) individuals. The overall prevalence of wheeze was similar in those living within 150 m of a road compared to those living further away (3.9% v 3.7%), but among the 3592 individuals living within 150 m, the risk of wheeze increased significantly in linear relation to proximity to the road (adjusted odds ratio = 1.17 per 30 m proximity, 95% CI 1.01 to 1.36). This relation was stronger, though not significantly so, for roads with above median traffic flows. CONCLUSION: These findings indicate that living in close proximity to road vehicle traffic is associated with an increased risk of wheeze, but that other environmental factors are also likely to be important. PMID- 15901885 TI - Non-malignant consequences of decreasing asbestos exposure in the Brazil chrysotile mines and mills. AB - AIMS: To investigate the consequences of improvement in the workplace environment over six decades (1940-96) in asbestos miners and millers from a developing country (Brazil). METHODS: A total of 3634 Brazilian workers with at least one year of exposure completed a respiratory symptoms questionnaire, chest radiography, and a spirometric evaluation. The study population was separated into three groups whose working conditions improved over time: group I (1940-66, n = 180), group II (1967-76, n = 1317), and group III (1977-96, n = 2137). RESULTS: Respiratory symptoms were significantly related to spirometric abnormalities, smoking, and latency time. Breathlessness, in particular, was also associated with age, pleural abnormality and increased cumulative exposure to asbestos fibres. The odds ratios (OR) for parenchymal and/or non-malignant pleural disease were significantly lower in groups II and III compared to group I subjects (0.29 (0.12-0.69) and 0.19 (0.08-0.45), respectively), independent of age and smoking status. Similar results were found when groups were compared at equivalent latency times (groups I v II: 30-45 years; groups II v III: 20-25 years). Ageing, dyspnoea, past and current smoking, and radiographic abnormalities were associated with ventilatory impairment. Lower spirometric values were found in groups I and II compared to group III: lung function values were also lower in higher quartiles of latency and of cumulative exposure in these subjects. CONCLUSIONS: Progressive improvement in occupational hygiene in a developing country is likely to reduce the risk of non-malignant consequences of dust inhalation in asbestos miners and millers. PMID- 15901886 TI - Use of cellular telephones and brain tumour risk in urban and rural areas. AB - AIMS: To investigate the association between the use of cellular or cordless telephones and the risk for brain tumours in different geographical areas, urban and rural. METHODS: Patients aged 20-80 years, living in the middle part of Sweden, and diagnosed between 1 January 1997 and 30 June 2000 were included. One control matched for sex and age in five year age groups was selected for each case. Use of different phone types was assessed by a questionnaire. RESULTS: The number of participating cases was 1429; there were 1470 controls. An effect of rural living was most pronounced for digital cellular telephones. Living in rural areas yielded an odds ratio (OR) of 1.4 (95% CI 0.98 to 2.0), increasing to 3.2 (95% CI 1.2 to 8.4) with >5 year latency time for digital phones. The corresponding ORs for living in urban areas were 0.9 (95% CI 0.8 to 1.2) and 0.9 (95% CI 0.6 to 1.4), respectively. This effect was most obvious for malignant brain tumours. CONCLUSION: In future studies, place of residence should be considered in assessment of exposure to microwaves from cellular telephones, although the results in this study must be interpreted with caution due to low numbers in some of the calculations. PMID- 15901887 TI - Supermarket baker's asthma: how accurate is routine health surveillance? AB - BACKGROUND: Regular health surveillance is commonly recommended for workers exposed to occupational antigens but little is known about how effective it is in identifying cases. AIMS: To report one large company's surveillance and compare its findings with those of a standard cross-sectional survey in the same workforce. METHODS: A supermarket company with 324 in-store bakeries producing bread from raw ingredients conducted a three-stage health surveillance programme in around 3000 bakery employees. The first stage involved the administration of a simple respiratory questionnaire. If chest symptoms were present a second questionnaire focusing on their work relationship was administered. If positive a blood sample was requested for the measurement of specific IgE to flour and fungal alpha-amylase. The results were compared to an independent cross-sectional survey of employees in 20 of the company's stores. RESULTS: Two hundred and ninety nine (92%) of the company's bakeries took part in surveillance. The overall employee response for the first stage was 77%; a quarter of those with respiratory symptoms reported that they were work related. Seventy four (61%) of those with work related chest symptoms had a measurement of specific IgE to either flour or fungal alpha-amylase, of whom 30 (41%) had a positive result. Surveillance estimated that 1% of bakery employees (1% bakers, 2% managers, 0.6% confectioners) had work related symptoms with specific IgE. This compared with 4% (7.5% bakers, 3.3% managers, 0% confectioners) in the cross-sectional survey (n = 166, 93% response). CONCLUSION: Comparison with a standard cross-sectional survey suggests that routine surveillance can underestimate the workplace burden of disease. The reasons may include technical or resource issues and uncertainties over confidentiality or the perceived consequences of participation. More research needs to be done looking into the design and efficacy of surveillance in occupational asthma. PMID- 15901888 TI - Atrazine in municipal drinking water and risk of low birth weight, preterm delivery, and small-for-gestational-age status. AB - BACKGROUND: Atrazine is a herbicide used extensively worldwide. Bioassays have shown that it is embryotoxic and embryolethal. Evidence of adverse reproductive outcomes from exposure in the general population is sparse. AIMS: To evaluate the association between atrazine levels in municipal drinking water and the following adverse reproductive outcomes: increased risk of preterm delivery, low birth weight (LBW), and small-for-gestational-age (SGA) status. METHODS: A total of 3510 births that took place from 1 October 1997, to 30 September 1998 were analysed. Atrazine measurements were available for 2661 samples from water treatment plants over the past decade. A seasonal pattern was identified, with atrazine peaking from May to September. The geometric mean of the atrazine level for this period was calculated for each water distribution unit and merged with the individual data by municipality of residence. RESULTS: Atrazine levels in water were not associated with an increased risk of LBW or SGA status and were slightly associated with prematurity. There was an increased risk of SGA status in cases in which the third trimester overlapped in whole or in part with the May September period, compared with those in which the third trimester occurred totally from October to April (OR = 1.37, 95% CI 1.04 to 1.81). If the entire third trimester took place from May to September, the OR was 1.54 (95% CI 1.11 to 2.13). CONCLUSIONS: Low levels of atrazine, a narrow exposure range, and limitations in the exposure assessment partly explain the lack of associations with atrazine. Findings point to the third trimester of pregnancy as the potential vulnerable period for an increased risk of SGA birth. Exposures other than atrazine and also seasonal factors may explain the increased risk. PMID- 15901889 TI - Required competencies of occupational physicians: a Delphi survey of UK customers. AB - BACKGROUND: Occupational physicians can contribute to good management in healthy enterprises. The requirement to take into account the needs of the customers when planning occupational health services is well established. AIMS: To establish the priorities of UK employers, employees, and their representatives regarding the competencies they require from occupational physicians; to explore the reasons for variations of the priorities in different groups; and to make recommendations for occupational medicine training curricula in consideration of these findings. METHODS: This study involved a Delphi survey of employers and employees from public and private organisations of varying business sizes, and health and safety specialists as well as trade union representatives throughout the UK. It was conducted in two rounds by a combination of computer assisted telephone interview (CATI) and postal survey techniques, using a questionnaire based on the list of competencies described by UK and European medical training bodies. RESULTS: There was broad consensus about the required competencies of occupational physicians among the respondent subgroups. All the competencies in which occupational physicians are trained were considered important by the customers. In the order of decreasing importance, the competencies were: Law and Ethics, Occupational Hazards, Disability and Fitness for Work, Communication, Environmental Exposures, Research Methods, Health Promotion, and Management. CONCLUSION: The priorities of customers differed from previously published occupational physicians' priorities. Existing training programmes for occupational physicians should be regularly reviewed and where necessary, modified to ensure that the emphasis of training meets customer requirements. PMID- 15901890 TI - Horse rug lung: toxic pneumonitis due to fluorocarbon inhalation. AB - Fluorocarbons are widely used in industry, and manifestations of inhalation toxicity include polymer fume fever, reactive airways dysfunction, and bronchospasm. Only seven cases of alveolitis occurring acutely after inhalation have been reported. This paper presents four cases of toxic pneumonitis due to direct inhalation of industrial fluorocarbon used as a waterproofing spray for horse rugs. These cases differ from previous reports and show that chronic as well as acute alveolitis can result from fluorocarbon inhalation. Corticosteroid treatment may be beneficial. The need for stricter control in the workplace is emphasised. PMID- 15901891 TI - World at work: Marble quarrying in Tuscany. PMID- 15901892 TI - Respiratory protective equipment. PMID- 15901893 TI - Pediatric disorders with autonomic dysfunction: what role for PHOX2B? AB - Hirschsprung disease, neuroblastomas, and congenital central hypoventilation syndrome can occur in combination, and familial cases have been reported in all three conditions. This suggests variable expression of a single genetic abnormality as the common cause to these neural crest disorders. Because the PHOX2B gene is pivotal in the development of most relays of the autonomic nervous system, including all autonomic neural crest derivatives, it was considered a candidate gene for the above conditions. Recent studies have shown that 1) PHOX2B is the main disease-causing gene for congenital central hypoventilation syndrome, an autosomal dominant disorder with incomplete penetrance; 2) PHOX2B is the first gene for which germline mutations have been demonstrated to predispose to neuroblastoma; and 3) Hirschsprung disease was associated with an intronic single nucleotide polymorphism of the PHOX2B gene in a case-control study. For clarifying the variable clinical expression of the autonomic nervous system dysfunction observed in neural crest disorders, international databases of clinical symptoms and molecular test results should be established. Furthermore, the development of genetic mouse models should help to improve our understanding of the molecular mechanisms underlying neural crest disorders. PMID- 15901894 TI - Murine teratology of fluconazole: evaluation of developmental phase specificity and dose dependence. AB - The potential of in utero exposure to fluconazole to initiate teratogenesis was analyzed in ICR (CD-1) mice. Developmental phase specificity was determined by treating mice with single oral doses of 700 mg/kg on gestational day 8, 9, 10, 11, or 12. Control animals received vehicle on gestational days 8-12. Gestational day 10 was identified as the phase of maximal sensitivity for induction of cleft palate, the predominant teratogenic effect induced by fluconazole, with 50% of fetuses exposed on this developmental phase being affected. After treatments on gestational day 8, 9, 11, or 12, cleft palate occurred with lower frequencies: 12, 21, 28.7, and 2.7%, respectively. Examination of skeletal morphology revealed anomalies of the middle ear apparatus in 15% of the fetuses that were exposed on gestational day 8. Dysmorphic tympanic ring and absence of the incus were the more common ear anomalies recorded. Reduced humeral length was noted in 22% of fetuses that were exposed on gestational day 10. Dose-response relationship was investigated by treating animals with 0 (vehicle), 87.5, 175, or 350 mg/kg on gestational day 10, coincident with the phase of peak teratogenic sensitivity. Besides showing that fluconazole operates under a strict dose-response mechanism, the study identified 175 mg/kg as the lowest observed adverse effect level for cleft palate induction, with 7.6% of the exposed fetuses being affected. PMID- 15901895 TI - Plasminogen activator inhibitor-1 and tissue-plasminogen activator in minority adolescents with type 2 diabetes and obesity. AB - Increased plasminogen activator inhibitor-1 (PAI-1) and decreased tissue plasminogen activator (t-PA) activities lead to impaired fibrinolysis, which is critical for cardiovascular disease. We studied these hemostatic factors at fasting state and after an oral fat load in 12 type 2 diabetic and 17 nondiabetic obese adolescents, matched for age, sex, body mass index, and sexual maturation. Plasma PAI-1, t-PA, and glucose as well as serum C-peptide, insulin, total cholesterol, triglyceride, and HDL and LDL cholesterol levels were measured at 0, 2, 4, and 6 h after the fat load. Metabolic responses were expressed as the area under the curve (AUC). PAI-1 activities were significantly greater in patients than in control subjects [fasting, 23.4 +/- 2.6 versus 12.9 +/- 2.0 U/mL (p < 0.004); AUC, 101.7 +/- 12.1 versus 57.6 +/- 6.5 U . h [corrected] . mL(-1) (p < 0.003)]. Fasting t-PA activities were significantly lower in the patients than in the control subjects (0.8 +/- 0.3 versus 6.5 +/- 2.7 U/mL; p < 0.001). Triglyceride was the only lipid parameter that was significantly different in the patients than in the control subjects [fasting, 1.5 +/- 0.2 versus 0.9 +/- 0.1 mM (p < 0.05); AUC, 15.7 +/- 2.9 versus 7.9 +/- 0.6 mmol . h(-1) . L(-1) (p < 0.02)]. The PAI-1 activities decreased significantly during the loading tests (p < 0.0001), whereas the t-PA activities did not change. Insulin resistance estimated by the homeostasis model assessment was greater in the patients than in the control subjects (14.4 +/- 2.8 versus 4.6 +/- 0.7; p < 0.0001). We conclude that elevated PAI-1 and diminished t-PA activities, suggestive of suppressed fibrinolysis, are present in our adolescents with type 2 diabetes; adding another risk factor for cardiovascular disease and acute high fat load does not further negatively affect this suppressed fibrinolysis. PMID- 15901896 TI - Guanylin and E. coli heat-stable enterotoxin induce chloride secretion through direct interaction with basolateral compartment of rat and human colonic cells. AB - We previously detected specific binding activity of Escherichia coli heat-stable enterotoxin (ST), the guanylin exogenous ligand, in rat colonic basolateral membranes. Because guanylin circulates in the bloodstream, we tested the hypothesis that it modulates intestinal ion transport by acting on the serosal side of intestinal cells. The effects of the mucosal and serosal addition of ST and guanylin on ion transport were investigated in the rat proximal colon and in Caco-2 cells in Ussing chambers, by monitoring short-circuit current (Isc). cGMP concentration was measured in Caco-2 cells by RIA. Mucosal ST addition induced an increase in Isc in rat proximal colon consistent with anion secretion. Serosal addition induced the same effects but to a lesser extent. The electrical effects observed in Caco-2 cells paralleled those observed in rat proximal colon. A pattern similar to the electrical response was observed with cGMP concentration. Guanylin addition to either side of Caco-2 cells induced the same effects as ST, although to a lesser extent. In all conditions, the electrical effect disappeared in the absence of chloride. ST directly interacts with basolateral receptors in the large intestine inducing chloride secretion through an increase of cGMP. However, the serosal effects are less pronounced compared with those observed with mucosal addition. Guanylin shows the same pattern, suggesting that it plays a role in the regulation of ion transport in the colon, but the relative importance of serosally mediated secretion remains to be determined. PMID- 15901897 TI - Carbonic anhydrase XIV is enriched in specific membrane domains of retinal pigment epithelium, Muller cells, and astrocytes. AB - Carbonic anhydrases (CAs) are ubiquitous enzymes important to many cell types throughout the body. They help determine levels of H(+) and HCO(-)(3) and thereby regulate intracellular and extracellular pH and volume. CA XIV, an extracellular membrane-bound CA, was recently shown to be present in brain and retina. Here, we analyze the subcellular distribution of CA XIV in retina by high-resolution immunogold cytochemistry and show that the distribution in retina (on glial cells but not neurons) is different from that reported for brain (on neurons but not glia). In addition, CA XIV is strongly expressed on retinal pigment epithelium (RPE). The specific membrane domains that express CA XIV were endfoot and nonendfoot membranes on Muller cells and astrocytes and apical and basolateral membranes of RPE. Gold particle density was highest on microvilli plasma membranes of RPE, where it was twice that of glial endfoot and Muller microvilli membranes and four times that of other glial membrane domains. Neither neurons nor capillary endothelial cells showed detectable labeling for CA XIV. This enrichment of CA XIV on specific membrane domains of glial cells and RPE suggests specialization for buffering pH and volume in retinal neurons and their surrounding extracellular spaces. We suggest that CA XIV is the target of CA inhibitors that enhance subretinal fluid absorption in macular edema. In addition, CA XIV may facilitate CO(2) removal from neural retina and modulate photoreceptor function. PMID- 15901898 TI - Activated antigen-presenting cells select and present chemically modified peptides recognized by unique CD4 T cells. AB - CD4 T cells recognized posttranslationally modified peptides of the protein hen egg-white lysozyme (HEL), consisting of nitration of tyrosines and modifications of tryptophans in the T cell contact residues of the peptides. T cells were directed against modifications of a chemically dominant HEL peptide as well as a minor HEL peptide, bound to the class II histocompatibility molecule I-A(k). The modified peptides were generated in vivo after immunization with native HEL molecules or were generated ex vivo by peroxynitrite treatment of HEL. Moreover, antigen-presenting cells (APC), either macrophages or dendritic cells activated in culture or in vivo, generated the modified HEL epitopes that stimulated the T cells. In transgenic mice expressing HEL, the T cells to the modified epitopes escaped negative selection and were found, albeit fewer in number than in normal mice. Infection with Listeria monocytogenes of the transgenic HEL mice generated APC containing the modifications. T cells to modified epitopes induced by activation of APC may be a component of antimicrobial immunity and autoimmune reactions. PMID- 15901899 TI - P2Y1 receptor signaling is controlled by interaction with the PDZ scaffold NHERF 2. AB - P2Y(1) purinergic receptors (P2Y(1)Rs) mediate rises in intracellular Ca(2+) in response to ATP, but the duration and characteristics of this Ca(2+) response are known to vary markedly in distinct cell types. We screened the P2Y(1)R carboxyl terminus against a recently created proteomic array of PDZ (PSD-95/Drosophila Discs large/ZO-1 homology) domains and identified a previously unrecognized, specific interaction with the second PDZ domain of the scaffold NHERF-2 (Na(+)/H(+) exchanger regulatory factor type 2). Furthermore, we found that P2Y(1)R and NHERF-2 associate in cells, allowing NHERF-2-mediated tethering of P2Y(1)R to key downstream effectors such as phospholipase Cbeta. Finally, we found that coexpression of P2Y(1)R with NHERF-2 in glial cells prolongs P2Y(1)R mediated Ca(2+) signaling, whereas disruption of the P2Y(1)R-NHERF-2 interaction by point mutations attenuates the duration of P2Y(1)R-mediated Ca(2+) responses. These findings reveal that NHERF-2 is a key regulator of the cellular activity of P2Y(1)R and may therefore determine cell-specific differences in P2Y(1)R-mediated signaling. PMID- 15901900 TI - Differential adhesion of amino acids to inorganic surfaces. AB - A fundamental, yet underexplored, materials system is the interface between biological molecules and inorganic surfaces. In an elemental approach to this problem, we have systematically examined the adhesion of amino acids to a series of inorganic surfaces including metals, insulators, and semiconductors. Significant differential adhesion is observed over the full complement of amino acids, determined largely by amino acid side-chain charge. Extensive mapping of the amino acid adhesion versus materials in multiple solutions is presented, with preliminary mechanisms derived from concentration and pH dependence. These results provide an empirical basis for building peptide to inorganic surface structures, and, using this adhesion data, we design inorganic nanostructures that are shown to selectively bind to prescribed primary peptide sequences. PMID- 15901901 TI - Radiographic validation of the Manchester scale for the classification of hallux valgus deformity. AB - OBJECTIVES: Hallux valgus is a common orthopaedic condition affecting elderly people. Grading the severity of the condition commonly involves obtaining measurements from radiographs, which may not be feasible or necessary in some clinical or research settings. Recently, a non-invasive clinical assessment tool (the Manchester scale), consisting of four standardized photographs, has been developed; however, its validity has not yet been determined. Therefore, the objective of this study was to determine the validity of this tool by correlating Manchester scale scores with hallux valgus measurements obtained from radiographs. METHODS: Weight-bearing dorsoplantar foot radiographs were obtained from 95 subjects (31 men and 64 women) aged 62-94 yr (mean 78.6, s.d. 6.5), and measurements of the hallux abductus angle, intermetatarsal angle and hallux interphalangeal adbuctus angle were performed. These measurements were then correlated with the Manchester scale scores (none, mild, moderate or severe). RESULTS: The Manchester scale score was highly correlated with hallux abductus angle (Spearman's rho = 0.73, P<0.01) and moderately associated with intermetatarsal angle (rho = 0.49, P<0.01) measurements obtained from radiographs. Analysis of variance revealed significant differences in mean hallux abductus angles [F3 = 119.99, P<0.001] and intermetatarsal angles [F3 =29.56, P<0.001] between the four Manchester scale categories. CONCLUSIONS: These findings indicate that the Manchester scale provides a valid representation of the degree of hallux valgus deformity determined from radiographic measurement of hallux abductus angle and intermetatarsal angle. We therefore recommend the use of this instrument as a simple, non-invasive screening tool for clinical and research purposes. PMID- 15901902 TI - The clinical and genetic associations of anti-cyclic citrullinated peptide antibodies in psoriatic arthritis. AB - OBJECTIVES: Antibodies recognizing a cyclic citrullinated peptide (anti-CCP) are highly specific for rheumatoid arthritis (RA) but their role in psoriatic arthritis (PsA) remains unclear. The aim of this study was therefore to investigate the prevalence of anti-CCP antibodies in PsA and assess their clinical and genetic associations. METHODS: One hundred and twenty-six patients with PsA, 40 patients with seropositive RA and 40 controls were tested for the presence of anti-CCP antibodies, rheumatoid factor (RF) and the HLA-DRB1 shared epitope. Clinical and radiological data were collected prospectively on all patients and compared between anti-CCP-positive and -negative patients. RESULTS: Seven (5.6%) patients with PsA were positive for anti-CCP antibodies compared with 0% of controls and 97% of patients with seropositive RA. The presence of anti-CCP antibodies in PsA was significantly associated with the HLA-DRB1 shared epitope (P<0.005), erosive disease (P<0.05), number of swollen joints (P<0.02) and DMARD use (P<0.05). CONCLUSIONS: Overall, the increased prevalence of anti CCP antibodies in this PsA population failed to reach statistical significance. However, when present, they were a marker of disease severity and had RA-linked MHC class II associations. Further studies are needed in a larger population of patients with PsA and appropriate controls to confirm any true association that may be present. PMID- 15901903 TI - Methotrexate-induced pancytopenia: serious and under-reported? Our experience of 25 cases in 5 years. AB - OBJECTIVE: To ascertain the extent of methotrexate (MTX)-related pancytopenia at the Norfolk and Norwich University Hospital (NNUH) between 1999 and 2004. METHODS: Patients were identified by a department database search, review of pharmacy records and personal communication. Pancytopenia was defined as white blood cell count (WBC) <3.5 x 10(9)/l, haemoglobin (Hb) <11 g/dl and platelet count <130 x 10(9)/l. Severe pancytopenia was defined as WBC <2.0 x 10(9)/l, Hb <10 g/dl and platelet count <50 x 10(9)/l. RESULTS: Twenty-five patients had MTX induced pancytopenia. Eleven patients were taking folic acid and one folinic acid. The median dose of MTX was 12.5 mg weekly (interquartile range 5.625 mg) and median duration of treatment 36 months (interquartile range 40.5 months). The severity of pancytopenia correlated with the dose (P = 0.04). The numbers of patients with potential risk factors were: renal insufficiency, 8; pre-existing folate deficiency, 7; age >75 yr, 15; hypoalbuminaemia, 18; pre-existing infection with hip prosthesis, 1; possible drug interactions, 18; dosing errors, 1; and polypharmacy, 15. Pancytopenia was detected by routine blood monitoring in nine patients. There were seven deaths (28% mortality), five from sepsis and two from acute myeloid leukaemia. CONCLUSION: This is the largest reported individual case series of MTX-induced pancytopenia. With the increasing long-term use of MTX, it is important that patients be monitored for haematological side-effects as pancytopenia can be a late manifestation. Pharmacogenetics may hold the answer to predicting who is at risk of this potentially fatal complication of MTX. PMID- 15901904 TI - BSR guidelines for prescribing TNF-alpha blockers in adults with ankylosing spondylitis. Report of a working party of the British Society for Rheumatology. PMID- 15901905 TI - Parathyroid hormone-related peptide expression in rat collagen-induced arthritis. AB - OBJECTIVES: The aim of this study was to describe expression of parathyroid hormone-related peptide (PTHrP) in collagen-induced arthritis (CIA), a well established animal model for rheumatoid arthritis. METHODS: CIA was induced in female dark agouti rats. Inguinal (ILNs) and popliteal (PLNs) lymph nodes and distal interphalangeal joints (DIP) were retrieved at different time points. Tissues were processed for detection of PTHrP and cell marker proteins by immunohistochemistry. Lymph node RNA was extracted, and PTHrP mRNA quantified using competitive reverse transcriptase polymerase chain reaction. RESULTS: Hyperplasia of ILNs was observed 2 days after injection, coinciding with the peak in PTHrP expression in ILNs (1240 +/- 373 gene copies/ng RNA vs normal 339 +/- 120, P < 0.05). Hyperplasia of PLNs was first seen at 1 day after onset of arthritis, coinciding with the peak in PTHrP expression in PLNs (2267 +/- 697 vs normal 781 +/- 136, P < 0.01). PTHrP expression in PLNs remained increased 5 days after onset (1361 +/- 302 vs normal 781 +/- 136, P < 0.05). In both PLNs and ILNs PTHrP protein was localized to high endothelial venules, lymphocytes and monocytes/macrophages. In DIP joint synovium PTHrP staining was first detected on day 10 after onset, and was most abundant at day 20 after onset, at sites of bone resorption and deposition, where it was localized to neutrophils, cells of monocyte lineage and osteoblasts. CONCLUSIONS: Changes in ILN and PLN PTHrP mRNA expression suggest that elevated levels of the cytokine are associated with aggravation of the inflammatory immune response. Changes in PTHrP in DIP joints indicate its involvement in late rather than early pathogenic events in CIA joints. PMID- 15901906 TI - Cytokine genotypes correlate with pain and radiologically defined joint damage in patients with juvenile rheumatoid arthritis. AB - OBJECTIVES: Single nucleotide polymorphisms (SNPs) in cytokine genes have been associated with risk of a number of autoimmune diseases. Moreover, some SNPs are associated with variations in rates of in vitro gene expression, and it is therefore possible that these functional polymorphisms may differentially affect inflammatory processes and disease outcome. This project's objective was to determine whether cytokine genotypes correlate with disease outcomes in patients with juvenile rheumatoid arthritis (JRA). METHODS: Genotypes of SNPs of pro inflammatory cytokines, tumour necrosis factor-alpha -308G -->A, interleukin-6 (IL-6) -174G -->C and interferon-gamma +874G -->A, and anti-inflammatory, immunosuppressive cytokines, interleukin-10 -1082G -->A, -819C -->T and -592A - >C and transforming growth factor-beta1 (TGF-beta1) codon 10T -->C and codon 25G ->C, were determined for patients with JRA who previously participated in a long term outcome study. Cytokine genotypes and clinical variables showing significant correlations with clinical outcomes at the alpha = 0.100 level in univariate analyses were entered in multivariate tests. RESULTS: In multivariate tests, the IL-6 genotype -174G/G was positively correlated with pain [regression coefficient B = 0.899, 95% confidence intervals (CI) 0.185, 1.612, P = 0.014]. The homozygous TGF-beta1 codon 25G/G genotype showed a protective effect against joint space narrowing on radiographs taken within 2 yr of disease onset, but confidence intervals were wide [odds ratio (OR) 0.176, 95% CI 0.037, 0.837 P = 0.029]. CONCLUSIONS: The correlation of IL-6 genotype with pain and the possible association of the TGF-beta1 codon 25 genotype with short-term radiographic damage (G/C with greater risk and G/G with decreased risk) suggests that both these polymorphisms may be useful early prognostic indicators. Further studies of the relation between cytokine genotypes and outcomes in patients with all forms of juvenile idiopathic arthritis (JIA) are warranted. PMID- 15901907 TI - Key autoantigens in SLE. PMID- 15901908 TI - Spring forward. PMID- 15901909 TI - MiMage: a Pan-European project on the role of mitochondria in aging. AB - The new European research project MiMage, supported by the European Community's Sixth Framework for Research and Technological Development, focuses on elucidating the role of mitochondria in conserved mechanisms of aging. Expertise in different research areas, including biochemistry, cell biology, genetics, molecular biology, and physiology, is provided by twelve research teams from seven European countries, together with one associated team from Canada and the United States. This report provides an introduction to the participating laboratories and the topics that will be addressed within the project, together with a concise report on the first symposium on the role of mitochondria in aging. PMID- 15901910 TI - Biphasic lindane-induced oxidation of glutathione and inhibition of gap junctions in myometrial cells. AB - The insecticide lindane (gamma-hexachlorocyclohexane) inhibits gap junction intercellular communication in rat myometrial cells by a mechanism involving oxidative stress. We hypothesized that oxidation of reduced glutathione (GSH) to glutathione disulfide (GSSG) and subsequent S-glutathionylation provide a mechanistic link between lindane-induced oxidative stress and lindane's inhibition of myometrial gap junction communication. Gap junction communication between cultured rat myometrial myocytes was assessed by Lucifer yellow dye transfer after microinjection. A biphasic pattern was confirmed, with dye transfer nearly abolished after 1 h of exposure to 100 microM lindane followed initially by recovery after lindane removal, and then the development 4 h after termination of lindane exposure of a delayed-onset, sustained inhibition that continued for 96 h. As measured by HPLC, cellular GSH varied over a 24-h period in a biphasic fashion that paralleled lindane-induced inhibition of dye transfer, whereas GSSG levels increased in a manner inversely related to GSH. In accordance, GSH/GSSG ratios were depressed at times when GSH and dye transfer were low. Lindane substantially increased S-glutathionylation in a concentration dependent manner, measured biochemically by GSSG reductase-stimulated release of GSH from precipitated proteins. Furthermore, treatments that promoted accumulation of GSSG (50 microM diamide and 25 microM 1,3-bis(2-chloroethyl)-1 nitrosourea [BCNU]) inhibited Lucifer yellow dye transfer between myometrial cells. Findings that lindane induced GSH oxidation to GSSG with increased S glutathionylation, together with the diamide and BCNU results, suggest that oxidation of GSH to GSSG is a component of the mechanism by which lindane inhibits myometrial gap junctions. PMID- 15901911 TI - Gene ontology mapping as an unbiased method for identifying molecular pathways and processes affected by toxicant exposure: application to acute effects caused by the rodent non-genotoxic carcinogen diethylhexylphthalate. AB - Toxicogenomics has the potential to reveal the molecular pathways and cellular processes that mediate the adverse responses to a toxicant. However, the initial output of a toxicogenomic experiment often consists of large lists of genes whose expression is altered after toxicant exposure. To interpret gene expression changes in the context of underlying biological pathways and processes, new bioinformatics methods must be developed. We have used global gene expression profiling combined with an evaluation of Gene Ontology (GO) and pathway mapping tools as unbiased methods for identifying the molecular pathways and processes affected upon toxicant exposure. We chose to use the acute effects caused by the non-genotoxic carcinogen and peroxisome proliferator (PP) diethylhexylphthalate (DEHP) in the mouse liver as a model system. Consistent with what is known about the mode of action of DEHP, our GO analysis of transcript profiling data revealed a striking overrepresentation of genes associated with the peroxisomal cellular component, together with genes involved in carboxylic acid and lipid metabolism. Furthermore we reveal gene expression changes associated with additional biological functions, including complement activation, hemostasis, the endoplasmic reticulum overload response, and circadian rhythm. Together, these data reveal potential new pathways of PP action and shed new light on the mechanisms by which non-genotoxic carcinogens control hepatocyte hypertrophy and proliferation. We demonstrate that GO mapping can identify, in an unbiased manner, both known and novel DEHP-induced molecular changes in the mouse liver and is therefore a powerful approach for elucidating modes of toxicity based on toxicogenomic data. PMID- 15901912 TI - Pronounced synergistic promotion of N-bis(2-hydroxypropyl)nitrosamine-initiated thyroid tumorigenesis in rats treated with excess soybean and iodine-deficient diets. AB - We have reported that excess soybean treatment and iodine deficiency synergistically interact, resulting in remarkable induction of thyroid hyperplasias in rats. In the present study, modifying effects of excess soybean and iodine-deficient diets were investigated in the post-initiation phase of N bis(2-hydroxypropyl)nitrosamine [DHPN]-initiated thyroid tumorigenesis in rats. AIN-93G in which casein was replaced with gluten was used as a basal diet to avoid possible iodine contamination. In Experiment 1, F-344 rats of both sexes were sc injected with DHPN at a dose of 2800 mg/kg body weight and then fed a diet containing 0%, 0.8%, 4%, or 20% defatted soybean for 12 weeks, with proportional replacement of gluten by soybean flour. Although no thyroid proliferative lesions were found in any group, the absolute thyroid weights were significantly (p < 0.01) elevated with the 20% soybean treatment. In Experiment 2, after similar sc injection of DHPN, rats were fed a basal diet or a diet containing 20% soybean under iodine normal or deficient conditions for 12 weeks. Soybean feeding to both sexes under iodine deficient but not normal conditions dramatically enhanced the development of thyroid follicular adenomas (p < 0.01) and adenocarcinomas (p < 0.05), in good agreement with decrease in thyroxine and increase in thyroid-stimulating hormone. Thus co-exposure to excess soybean and iodine deficiency results in synergistic promotion of DHPN-initiated thyroid tumorigenesis in rats, of which mechanisms appear to primarily involve effects on serum hormone levels. PMID- 15901913 TI - Nrf2 activation involves an oxidative-stress independent pathway in tetrafluoroethylcysteine-induced cytotoxicity. AB - Tetrafluoroethylcysteine (TFEC), a metabolite of the industrial gas tetrafluoroethylene, can cause both nephrotoxicity and limited hepatotoxicity in animal models, and this is associated with the covalent modification of specific intramitochondrial proteins including heat shock protein 60 (HSP60), mitochondrial HSP70 (mtHSP70), aspartate aminotransferase (AST), aconitase, and alpha-ketoglutarate dehydrogenase (alphaKGDH). Using the murine TAMH cell line as a useful in vitro model for TFEC toxicity, we demonstrate a rapid and sustained induction of Nrf2, a member of the "cap-and-collar" transcription factor family, following exposure to cytotoxic concentrations of TFEC. A functional correlate was also established with the rapid translocation of cytosolic Nrf2 into the nucleus. In addition, transcriptional and translational upregulation of known Nrf2 regulated genes including glutamate cysteine ligase (GCL), both catalytic and modulatory subunits, heme oxygenase-1, and glutathione S-transferase (GST) isoforms were detected. While Nrf2 activation is often linked to perturbation of cellular thiol status and/or oxidative stress, we were unable to detect any significant depletion of cellular glutathione or oxidation of mitochondrial membrane cardiolipin or increases in reactive oxygen species (ROS). These data suggest Nrf2 activation is likely independent of classical oxidative stress or, at best, a result of a transient, low-level redox stress. Moreover, supporting evidence indicates an early endoplasmic reticular (ER) stress response after TFEC treatment, with a time-dependent upregulation of the ER responsive genes gadd34, gadd45, gadd153, and ndr1 . These findings suggest an alternative pathway for Nrf2 activation, i.e., Nrf2 phosphorylation through ER-mediated protein kinases such as PKR-like endoplasmic reticular kinase (PERK). Overall, the results implicate a role for Nrf2 in the cellular response to TFEC toxicity and suggest a previously unrecognized role for the ER in this model of mitochondrially initiated cytotoxicity. PMID- 15901914 TI - Di-n-butyl phthalate activates constitutive androstane receptor and pregnane X receptor and enhances the expression of steroid-metabolizing enzymes in the liver of rat fetuses. AB - The plasticizer di-n-butyl phthalate (DBP) is a reproductive toxicant in rodents. Exposure to DBP in utero at high doses alters early reproductive development in male rats. Di-n-butyl phthalate also affects hepatic and extrahepatic enzymes. The objectives of this study were to determine the responsiveness of steroid metabolizing enzymes in fetal liver to DBP and to investigate the potential of DBP to activate nuclear receptors that regulate the expression of liver enzymes. Pregnant Sprague-Dawley rats were orally dosed with DBP at levels of 10, 50, or 500 mg/kg/day from gestation days 12 to 19; maternal and fetal liver samples were collected on day 19 for analyses. Increased protein and mRNA levels of CYP 2B1, CYP 3A1, and CYP 4A1 were found in both maternal and fetal liver in the 500-mg dose group. Di-n-butyl phthalate at high doses also caused an increase in the mRNA of hepatic estrogen sulfotransferase and UDP-glucuronosyltransferase 2B1 in the dams but not in the fetuses. Xenobiotic induction of CYP3A1 and 2B1 is known to be mediated by the nuclear hormone receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR). In vitro transcriptional activation assays showed that DBP activates both PXR and CAR. The main DBP metabolite, mono butyl-phthalate (MBP) did not interact strongly with either CAR or PXR. These data indicate that hepatic steroid- and xenobiotic-metabolizing enzymes are susceptible to DBP induction at the fetal stage; such effects on enzyme expression are likely mediated by xenobiotic-responsive transcriptional factors, including CAR and PXR. Our study shows that DBP is broadly reactive with multiple pathways involved in maintaining steroid and lipid homeostasis. PMID- 15901915 TI - Effects of atrazine on CYP19 gene expression and aromatase activity in testes and on plasma sex steroid concentrations of male African clawed frogs (Xenopus laevis). AB - Some investigators have suggested that the triazine herbicide atrazine can cause demasculinization of male amphibians via upregulation of the enzyme aromatase. Male adult African clawed frogs (Xenopus laevis) were exposed to three nominal concentrations of atrazine (1, 25, or 250 microg atrazine/l) for 36 days, and testicular aromatase activity and CYP19 gene expression, as well as concentrations of the plasma sex steroids testosterone (T) and 17beta-estradiol (E2), and gonad size (GSI) were measured. There were no effects on any of the parameters measured, with the exception of plasma T concentrations. Plasma T concentrations in X. laevis exposed to the greatest concentration of atrazine were significantly less (p = 0.034) than those in untreated frogs. Both CYP19 gene expression and aromatase activities were low regardless of treatment, and neither parameter correlated with the other. We conclude that aromatase enzyme activity and gene expression were at basal levels in X. laevis from all treatments, and that the tested concentrations of atrazine did not interfere with steroidogenesis through an aromatase-mediated mechanism of action. PMID- 15901916 TI - Effects of two fungicides with multiple modes of action on reproductive endocrine function in the fathead minnow (Pimephales promelas). AB - Many chemicals that adversely affect reproduction and/or development do so through multiple pathways within the reproductive tract and hypothalamic pituitary-gonadal axis. Notable in this regard are fungicides, such as prochloraz or fenarimol, which in mammals have the potential to impact endocrine function through inhibition of CYP enzymes involved in steroid metabolism, as well as through antagonism of the androgen receptor(s). The objective of our studies was to assess the effects of prochloraz and fenarimol on reproductive endocrine function in a model small fish species, the fathead minnow (Pimephales promelas), using both in vitro and in vivo assays. The two fungicides inhibited in vitro CYP19 aromatase activity in brain and ovarian homogenates from the fish, with prochloraz exhibiting a greater potency than fenarimol. Prochloraz and fenarimol also bound competitively to the cloned fathead minnow androgen receptor expressed in COS-1 cells. The two fungicides significantly reduced fecundity of the fish in a 21-day reproduction assay at water concentrations of 0.1 (prochloraz) and 1.0 (fenarimol) mg/l. The in vivo effects of prochloraz on plasma steroid (17beta estradiol, testosterone, 11-ketotestosterone) and vitellogenin (an estrogen responsive protein) concentrations, as well as on gonadal histopathology, were consistent with inhibition of steroidogenesis. Fenarimol also affected several aspects of endocrine function in vivo; however, the suite of observed effects did not reflect either aromatase inhibition or androgen receptor antagonism. These studies contribute to a better mechanistic understanding of the extrapolation of effects of endocrine-disrupting chemicals across vertebrate classes. PMID- 15901917 TI - Accumulation, tissue distribution, and maternal transfer of dietary 2,3,7,8, tetrachlorodibenzo-p-dioxin: impacts on reproductive success of zebrafish. AB - TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) is a reproductive toxicant and endocrine disruptor in nearly all vertebrates; however, the mechanisms by which TCDD alters the reproductive system is not well understood. The zebrafish provides a powerful vertebrate model system to investigate molecular mechanisms by which TCDD affects the reproductive system, but little is known regarding reproductive toxic response of zebrafish following chronic, sublethal exposure to TCDD. Here we investigate the accumulation of TCDD in selected tissues of adult female zebrafish and maternal transfer to offspring following dietary exposure to TCDD (0.08-2.16 ng TCDD/fish/day). TCDD accumulated in tissues of zebrafish in a dose- and time-dependent manner, except for brain. Chronic dietary exposure resulting in the accumulation of 1.1-36 ng/g fish did not induce an overt toxic response or suppress spawning activity. The ovosomatic index was impacted with an accumulation of as little as 0.6 ng/g fish, and 10% of the females showed signs of ovarian necrosis following accumulation of approximately 3 ng/g TCDD. Offspring health was impacted with an accumulation of as little as 1.1 ng/g female; thus the lowest observed effect level (LOEL) for reproductive toxicity in female zebrafish is approximately 0.6-1.1 ng/g fish. Maternal transfer resulted in the accumulation of 0.094-1.2 ng/g, TCDD, which was sufficient to induce the typical endpoints of larval TCDD toxicity, commonly referred to as blue sac syndrome. This study provides the necessary framework to utilize the zebrafish model system for further investigations into the molecular mechanisms by which TCDD exerts its reproductive toxic responses. PMID- 15901918 TI - Different types of combination effects for the induction of micronuclei in mouse lymphoma cells by binary mixtures of the genotoxic agents MMS, MNU, and genistein. AB - Distinction between dose addition and response addition for the analysis of the toxicity of mixtures may allow differentiation of the components regarding similar versus independent mode of action. For nonlinear dose responses for the components, curves of dose addition and response addition differ and embrace an "envelope of additivity." Synergistic or antagonistic interaction may then be postulated only if the mixture effect is outside this surface. This situation was analyzed for the induction of micronuclei in L5178Y mouse lymphoma cells by the two methylating agents methyl methanesulfonate (MMS) and N-methyl-N-nitrosourea (MNU) and the topoisomerase-II inhibitor genistein (GEN). All three chemicals reproducibly generated sublinear (upward convex) dose-response relationships. For the analysis of mixture effects, these genotoxic agents were investigated in the three binary combinations. Statistical testing for dose addition along parallel exponential dose responses was performed by linear regression with interaction based on the logarithm of the number of cells that contain micronuclei. For MMS+MNU, the mixture effect was compatible with dose addition (i.e., significantly larger than calculated for the addition of net responses). For MMS+GEN, the measured effect was larger than for response addition but smaller than for dose addition. For MNU+GEN, the measured effect was below response addition, indicative of true antagonism. In the absence of knowledge on the sublinear dose-response relationships for the individual components, a synergistic effect of MMS on both MNU and GEN would have been postulated erroneously. The observed difference between MMS and MNU when combined with GEN would not have been predicted on the basis of a simplistic interpretation of DNA methylation as the mode of action and may be due to differences in the profile of DNA methylations and/or epigenetic effects. We conclude that knowledge of nonlinearities of the dose-response curves of individual components of a mixture can be crucial to analyze for synergism or antagonism and that an in-depth mechanistic knowledge is useful for a prediction of similarity or independence of action. PMID- 15901919 TI - Neurobehavioral effects of chronic dietary and repeated high-level spike exposure to chlorpyrifos in rats. AB - This study aimed to model long-term subtoxic human exposure to an organophosphorus pesticide, chlorpyrifos, and to examine the influence of that exposure on the response to intermittent high-dose acute challenges. Adult Long Evans male rats were maintained at 350 g body weight by limited access to a chlorpyrifos-containing diet to produce an intake of 0, 1, or 5 mg/kg/day chlorpyrifos. During the year-long exposure, half of the rats in each dose group received bi-monthly challenges (spikes) of chlorpyrifos, and the other half received vehicle. Rats were periodically tested using a neurological battery of evaluations and motor activity to evaluate the magnitude of the acute response (spike days) as well as recovery and ongoing chronic effects (non-spike days). Effects of the spikes differed as a function of dietary level for several endpoints (e.g., tremor, lacrimation), and in general, the high-dose feed groups showed greater effects of the spike doses. Animals receiving the spikes also showed some neurobehavioral differences among treatment groups (e.g., hypothermia, sensory and neuromotor differences) in the intervening months. During the eleventh month, rats were tested in a Morris water maze. There were some cognitive deficits observed, demonstrated by slightly longer latency during spatial training, and decreased preference for the correct quadrant on probe trials. A consistent finding in the water maze was one of altered swim patterning, or search strategy. The high-dose feed groups showed more tendency to swim in the outer annulus or to swim very close to the walls of the tank (thigmotaxic behavior). Overall, dietary exposure to chlorpyrifos produced long lasting neurobehavioral changes and also altered the response to acute challenges. PMID- 15901920 TI - Gene expression changes induced in the testis by transplacental exposure to high and low doses of 17{alpha}-ethynyl estradiol, genistein, or bisphenol A. AB - The purpose of this study was to determine (1) the transcriptional program elicited by exposure to three estrogen receptor (ER) agonists: 17 alpha-ethynyl estradiol (EE), genistein (Ges), and bisphenol A (BPA) during fetal development of the rat testis and epididymis; and (2) whether very low dosages of estrogens (evaluated over five orders of magnitude of dosage) produce unexpected changes in gene expression (i.e., a non-monotonic dose-response curve). In three independently conducted experiments, Sprague-Dawley rats were dosed (sc) with 0.001-10 microg EE/kg/day, 0.001-100 mg Ges/kg/day, or 0.002-400 mg BPA/kg/day. While morphological changes in the developing reproductive system were not observed, the gene expression profile of target tissues were modified in a dose responsive manner. Independent dose-response analyses of the three studies identified 59 genes that are significantly modified by EE, 23 genes by Ges, and 15 genes by BPA (out of 8740), by at least 1.5 fold (up- or down-regulated). Even more genes were observed to be significantly changed when only the high dose is compared with all lower doses: 141, 46, and 67 genes, respectively. Global analyses aimed at detecting genes consistently modified by all of the chemicals identified 50 genes whose expression changed in the same direction across the three chemicals. The dose-response curve for gene expression changes was monotonic for each chemical, with both the number of genes significantly changed and the magnitude of change, for each gene, decreasing with decreasing dose. Using the available annotation of the gene expression changes induced by ER agonist, our data suggest that a variety of cellular pathways are affected by estrogen exposure. These results indicate that gene expression data are diagnostic of mode of action and, if they are evaluated in the context of traditional toxicological end-points, can be used to elucidate dose-response characteristics. PMID- 15901921 TI - Protein adduct formation as a molecular mechanism in neurotoxicity. AB - Chemicals that cause nerve injury and neurological deficits are a structurally diverse group. For the majority, the corresponding molecular mechanisms of neurotoxicity are poorly understood. Many toxicants (e.g., hepatotoxicants) of other organ systems and/or their oxidative metabolites have been identified as electrophiles and will react with cellular proteins by covalently binding nucleophilic amino acid residues. Cellular toxicity occurs when adduct formation disrupts protein structure and/or function, which secondarily causes damage to submembrane organelles, metabolic pathways, or cytological processes. Since many neurotoxicants are also electrophiles, the corresponding pathophysiological mechanism might involve protein adduction. In this review, we will summarize the principles of covalent bond formation that govern reactions between xenobiotic electrophiles and biological nucleophiles. Because a neurotoxicant can form adducts with multiple nucleophilic residues on proteins, the challenge is to identify the mechanistically important adduct. In this regard, it is now recognized that despite widespread chemical adduction of tissue proteins, neurotoxicity can be mediated through binding of specific target nucleophiles in key neuronal proteins. Acrylamide and 2,5-hexanedione are prototypical neurotoxicants that presumably act through the formation of protein adducts. To illustrate both the promise and the difficulty of adduct research, these electrophilic chemicals will be discussed with respect to covalent bond formation, suspected protein sites of adduction, and proposed mechanisms of neurotoxicity. The goals of future investigations are to identify and quantify specific protein adducts that play a causal role in the generation of neurotoxicity induced by electrophilic neurotoxicants. This is a challenging but critical objective that will be facilitated by recent advances in proteomic methodologies. PMID- 15901922 TI - An overview of chemosensitivity testing. AB - This overview chapter presents the importance of chemosensitivity testing for screening new therapeutic agents, identifying patterns of chemosensitivity for different types of tumors, establishing patterns of cross-resistance and sensitivity in treatment naive and relapsing tumors; identifying genomic and proteomic profiles associated with sensitivity; correlating in vitro response, preclinical in vivo effect, and clinical outcome associated with a particular therapeutic agent, and tailoring chemotherapy regimens to individual patients. Various assays are available to achieve these end points, including several in vitro clonogenic and proliferation assays, cell metabolic activity assays, molecular assays to monitor expression of markers for responsiveness, development of drug resistance and induction of apoptosis, in vivo tumor growth and survival assays in metastatic and orthotopic models, and in vivo imaging assays. The advantages and disadvantages of the specific assays are discussed. A summary of research areas related to chemo sensitivity testing is also included. PMID- 15901923 TI - Clonogenic cell survival assay. AB - The clonogenic cell survival assay determines the ability of a cell to proliferate indefinitely, thereby retaining its reproductive ability to form a large colony or a clone. This cell is then said to be clonogenic. A cell survival curve is therefore defined as a relationship between the dose of the agent used to produce an insult and the fraction of cells retaining their ability to reproduce. Although clonogenic cell survival assays were initially described for studying the effects of radiation on cells and have played an essential role in radiobiology, they are now widely used to examine the effects of agents with potential applications in the clinic. These include, in addition to ionizing radiation, chemotherapy agents such as etoposide and cisplatin, antiangiogenic agents such as endostatin and angiostatin, and cytokines and their receptors, either alone or in combination therapy. Survival curves have been generated for many established cell lines growing in culture. One can use cell lines from various origins including humans and rodents; these cells can be neoplastic or normal. Because survival curves have wide application in evaluating the reproductive integrity of different cells, we provide here the steps involved in setting up a typical experiment using an established cell line in culture. PMID- 15901924 TI - High-sensitivity cytotoxicity assays for nonadherent cells. AB - High-sensitivity cytotoxicity assays refer to assays that can detect high levels of cell kill, to many powers of 10, and that can detect, ideally, a single remaining viable cell. Two such assays are described here, which have been used with Raji B-lymphoma cells, and are applicable to other nonadherent target cells. The first is a cell-counting assay, performed over a 3-wk period, which provides a simple, reliable, and sensitive assay of cytotoxicity. By determining the time required for 16-fold multiplication, the apparent fraction surviving can be calculated. This assay does not correct for treatment-induced delays in cell division and is dependent on maintaining the cells in exponential growth. The second assay measures colony-forming units using a limiting dilution method. Feeder cells are required to obtain a high cloning efficiency. Each dilution is plated in 48 wells of a 96-well plate, and positive wells are scored rapidly, by eye, after two wk. PMID- 15901925 TI - Sulforhodamine B assay and chemosensitivity. AB - The sulforhodamine B (SRB) assay was developed by Skehan and colleagues to measure drug-induced cytotoxicity and cell proliferation for large-scale drug screening applications. Its principle is based on the ability of the protein dye sulforhodamine B to bind electrostatically and pH dependent on protein basic amino acid residues of trichloroacetic acid-fixed cells. Under mild acidic conditions it binds to and under mild basic conditions it can be extracted from cells and solubilized for measurement. Results of the SRB assay were linear with cell number and cellular protein measured at cellular densities ranging from 1 to 200% of confluence. Its sensitivity is comparable with that of several fluorescence assays and superior to that of Lowry or Bradford. The signal-to noise ratio is favorable and the resolution is 1000-2000 cells/well. It performed similarly compared to other cytotoxicity assays such as MTT or clonogenic assay. The SRB assay possesses a colorimetric end point and is nondestructive and indefinitely stable. These practical advances make the SRB assay an appropriate and sensitive assay to measure drug-induced cytotoxicity even at large-scale application. PMID- 15901926 TI - Use of the differential staining cytotoxicity assay to predict chemosensitivity. AB - The differential staining cytotoxicity (DiSC) assay is one of the total cell-kill assays that can be used for drug resistance testing. Numerous publications have demonstrated the clinical value of chemosensitivity testing with that assay (and similar ones). The DiSC assay is successful in the majority of malignant samples of which the cells can be brought in suspension (not necessarily as single-cell suspension). Although the assay is laborious and requires skilled technicians, it requires few cells, can be used for proliferating and nonproliferating cell populations, and can discriminate between malignant and contaminating nonmalignant cells. The latter is a major advantage of the DiSC assay. This chapter describes the practical aspects of this assay, several topics that need to be taken into account, and potential pitfalls. As such, it is not an extensive review of studies in which the DiSC assay was used. PMID- 15901927 TI - Collagen gel droplet culture method to examine in vitro chemosensitivity. AB - For effective cancer chemotherapy, chemosensitivity testing of anticancer drugs should be performed using fresh surgical specimens obtained from the cancer. We have developed a new in vitro chemosensitivity test named the collagen gel droplet embedded culture drug sensitivity test (CD-DST). The CD-DST method consists of a collagen gel droplet embedded culture step, exposure and washout of anticancer drug, a serum-free culture step, and evaluation of anticancer effect by image analysis. This method has many advantages including a high success rate for primary culture, the need for only a small number of cells for the test, easy quantification of the anticancer effects without contamination with fibroblasts by using an image analysis system, a good correlation between in vitro and in vivo results, and simplicity and speed. The CD-DST method can be performed in the laboratory using a system kit Primaster. PMID- 15901928 TI - The MTT assay to evaluate chemosensitivity. AB - The assessment of the degree or rate of cellular proliferation and cell viability is critical to the assessment of the effects of drugs, antibodies, or cytokines on both normal and malignant cell populations. This can be accomplished by either direct or indirect counting methods. Direct counting by manual or automated methods, using a hemacytometer or particle counter, respectively, allows for serial cell counting at multiple time points, but these are low-throughput approaches. High-throughput and robust alternatives to direct counting utilize either radiotracers (e.g., 3H-thymidine) or dye compounds, which can be adapted to multiwell culture plate formats. This chapter focuses on the use of tetrazolium-type indicator dyes, of which the compound 3-(4,5-dimethylthiazolyl 2)-2,5-diphenyltetrazolium bromide (MTT) has been the most widely utilized. Newer tetrazolium dyes that yield water-soluble products and offer added flexibility, increases in sensitivity, and fewer steps, which are offset by increased costs, are also covered. PMID- 15901929 TI - Histoculture drug response assay to monitor chemoresponse. AB - We provide a detailed explanation of the procedure of the histoculture drug response assay (HDRA) with 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) end point among several modified HDRA procedures. Fresh surgical specimens are cut into approx 1- to 2-mm3 pieces and put on a gelatin sponge infiltrated with culture medium containing a test drug. After incubation for 7 d, cell viability is assessed by the MTT assay. HDRA uses cancer tissue fragments with cells growing in three dimensions, with maintenance of intercellular contact and interactions with stromal cells. Therefore, it seems that HDRA can assess the sensitivity of tumor cells to anticancer drugs in conditions similar to those in vivo and, consequently, shows high prediction rate. PMID- 15901930 TI - In vitro testing of chemosensitivity in physiological hypoxia. AB - Highly aggressive, rapidly growing tumors are often hypoxic, owing to an inadequate supply relative to consumption of oxygen (O2) in the expanding tumor mass, or growth in tissues with physiologically low O2 concentrations (such as bone marrow). Selection of tumor cells that can grow or survive under hypoxic conditions appears from both experimental and clinical studies to impact tumor progression, response to therapy, and to increase resistance to radiation and to certain cytotoxic drugs. Therefore, the predictive value of preclinical testing of anticancer agents in cell culture might be improved by conducting testing in conditions of physiological hypoxia. We review the impact of hypoxia on anticancer drug cytotoxicity and the methods used in our laboratory to asses the cytotoxic activity of single antineoplastic drugs under conditions of physiological hypoxia. PMID- 15901931 TI - Chemosensitivity testing using microplate adenosine triphosphate-based luminescence measurements. AB - During the last two decades, novel nonclonogenic methods for pretherapeutic chemosensitivity testing have been developed that are likely to overcome major technical limitations of older assays such as low evaluability rates, low degree of standardization and reproducibility, lack of technical robustness, and poor methodological efficacy. Among these, the microplate adenosine triphosphate (ATP) based tumor chemosensitivity assay (ATP-TCA) has gained particular merits for ex vivo chemosensitivity testing of native nonhematological tumors including cancers of the breast, ovary, gastrointestinal tract, cervix and corpus uteri, and lung; malignant melanomas; gliomas; sarcomas; and mesotheliomas. For this indication, the ATP-TCA can now be considered the best documented and validated technology. This assay, which is now commercially available, provides a highly reproducible, easy-to-handle kit technique; low technical failure rates; and a high methodological efficacy requiring only 1 x 106 tumor cells to test four to six different drugs or combinations. In ovarian and breast carcinomas, the predictive accuracy is > 90%, with a positive predictive value of 85-90% and a negative predictive value near 100%, respectively. In primary ovarian cancers, the ATP-TCA has been found to accurately predict both clinical response and survival. In two prospective clinical trials in patients with heavily pretreated ovarian cancer, chemotherapy individually selected by the ATP-TCA has been found to triple the response rates and nearly double the survival compared to empirically chosen regimens. Consequently, this assay, which is now under phase III evaluation, has successfully been used in new agent development to screen for novel chemotherapy regimens for the treatment of patients with breast and ovarian carcinoma and melanoma, respectively. This chapter highlights the recent preclinical and clinical experience with this promising technology and gives a detailed description of all the technical aspects of the ATP-TCA. PMID- 15901932 TI - High-throughput technology: green fluorescent protein to monitor cell death. AB - Reliable assessment of cell death is now pivotal to many research programs aiming at generating new antitumor compounds or at screening cDNA libraries to identify genes with pro- or antiapoptotic functions. Such approaches need to rely on reproducible, easy handling, and rapid microplate-based cytotoxicity assays that are amenable to high-throughput screening technologies. We describe here a method for the direct measurement of cell death, based on the detection of a decrease in fluorescence observed following death induction in cells stably expressing enhanced green fluorescent protein (EGFP). Our data clearly show that such a decrease in EGFP fluorescence after cell death induction happens in various cell types, including those routinely used in anticancer drug screening (i.e., murine and human, lymphoid, fibroblastic, or epithelial cell lines). Moreover, the decrease in EGFP fluorescence is observed in cells induced to die by a variety of apoptosis-inducing agents, such as glucocorticoids (dexamethasone), DNA- damaging agents (etoposide, cisplatin), microtubule disorganizers (paclitaxel), protein kinase C inhibitors (staurosporine), or a caspase-independent apoptotic stimulus (CD45 crosslinking). A decrease in fluorescence can be assessed either by flow cytometry or with a fluorescence microplate reader. The kinetics and specificity of this EGFP-based assay were comparable with those of other conventional techniques used to detect cell death. This novel EGFP-based microplate assay combines sensitivity and rapidity and is amenable to high-throughput setups, making it an assay of choice for evaluation of cell cytotoxicity. PMID- 15901933 TI - DIMSCAN: a microcomputer fluorescence-based cytotoxicity assay for preclinical testing of combination chemotherapy. AB - DIMSCAN is a semiautomatic fluorescence-based digital image microscopy system that quantifies relative total (using a DNA stain) or viable (using fluorescein diacetate [FDA]) cell numbers in tissue culture multiwell plates ranging from 6 to 384 wells per plate. DIMSCAN is a rapid and efficient tool for conducting in vitro cytotoxicity assays across a 4 log dynamic range. The specificity of detecting viable cells with FDA is achieved by using digital image processing and chemical quenching of fluorescence in nonviable cells with eosin Y. Average scan time for the most commonly used format, a 96-well plate, is 6 min. Cytotoxicity for neuroblastoma cell lines measured by DIMSCAN was found to be comparable to manual Trypan blue dye exclusion counts or colony formation in soft agar, but with a significantly wider dynamic range, which enables drug combination studies used to detect synergistic or antagonistic interactions. The linearity of DIMSCAN was validated (r2 = 0.99967 +/- 0.0003) for cells stained with FDA deposited using a fluorescence-activated cell sorter, documenting a dynamic range > 4 logs, and the ability to detect a single viable cell in a well 93% of the time. DIMSCAN has been used to demonstrate preclinical activity of cytotostatic and cytotoxic drugs and drug combinations that have subsequently shown activity in clinical trials. PMID- 15901934 TI - The ChemoFx assay: an ex vivo cell culture assay for predicting anticancer drug responses. AB - We provide a detailed description of the ChemoFx Assay, a phenotype-based cell culture assay for predicting anticancer drug responses in individual cancer patients. The ChemoFx Assay is based on the outgrowth and short-term primary culture of epithelial cells derived from pieces of solid tumor adenocarcinomas that are obtained at the time of tumor resection. Malignant epithelial cells are grown attached in wells of microtiter plates and treated with six escalating doses of chemotherapeutic drug. Using an operator-controlled automated image analysis system, cell kill is measured microscopically by counting the number of live cells remaining after dead cells have detached and are subsequently rinsed away. A dose-response graph is automatically generated by comparing the number of cells in drug-treated wells with those in control wells. PMID- 15901935 TI - Evaluating response to antineoplastic drug combinations in tissue culture models. AB - The mainstay of clinical antineoplastic chemotherapy is multiagent combinations, most of which were developed empirically. Because of the desire to speed research and decrease costs, there is increasing interest in moving new drugs into clinical trials in potentially active combinations based on preclinical testing data. Different mathematical models have been proposed for evaluating drug interactions, which can be classified as synergistic (combinations demonstrating greater than the additive activity expected from each agent alone), additive, or antagonistic (drugs showing less activity in combination than expected from the sum of each agent alone). Here, we briefly review some of the principles for testing cytotoxic drug interactions. We focus this review on application of the Combination Index method (as developed by Chou and colleagues) in the evaluation of drug interactions in cell culture assays. PMID- 15901936 TI - Image analysis using the fluorochromasia assay to quantify tumor drug sensitivity. AB - A method of assessing chemosensitivity of tissue utilizing tissue fluorescence and image analysis was implemented to provide a rapid and quantitative means of assessing the effect of drugs on tissue metabolic activity and proliferative capacity. The fluorescent microscopic image captured by a silicon-intensified target (low-light-detecting) camera and linked to an image- processing unit was measured for fluorescent brightness and tumor image area. An established rodent model served to characterize the system's ability to measure serially the tumor's metabolic activity and growth. Further studies on fresh human tumors were conducted with a novel topoisomerase II inhibitor, NC-190. Tumor image area and fluorescent brightness were measured 24 h pretreatment, 48 h posttreatment, and 48 h post-drug removal. Fifty-five percent (28/51) of fresh human tumors showed sensitivity to 48-h exposure to 10, 30, or 100 microM NC-190. The potential benefit of this technique is the ability to predict the response of tumors to chemotherapeutic agents as a laboratory tool for preclinical drug evaluation and clinically prior to the commencement of therapy. PMID- 15901937 TI - Immunohistochemical detection of ornithine decarboxylase as a measure of chemosensitivity testing. AB - The development of reliable methods for the in vitro testing of sensitivity of cancer cells to various drugs has been a longstanding objective in cancer treatment. The development of individualized chemotherapy could minimize undesired toxic side effects and increase the chance of recovery. The known methods for in vitro chemosensitivity tests are mainly based on monitoring the metabolic changes induced in cancer cells by the drugs. These experiments, as well as attempts to cultivate isolated cancer cells, did not give reliable results. In this study, I used a marker for proliferation to detect the effect of drugs on the potential of cancer cells to divide. An ideal marker should be present in all cells, be expressed early in the cell cycle, and have a short half life. Ornithine decarboxylase (ODC), which catalyzes the conversion of ornithine to putrescine, fulfilled these prerequisites. Because ODC has an extremely short half-life, it disappears when cellular proliferation is arrested. The decay of ODC was assayed both by determining its activity and by immunohistochemical analyses. This approach was successfully used to determine the sensitivity of lymphocytes from hematological cancer patients to various drugs. It is conceivable that this method could serve as an important tool to improve cancer chemotherapy. PMID- 15901938 TI - Immunohistochemistry of p53, Bcl-2, and Ki-67 as predictors of chemosensitivity. AB - Chemosensitivity is affected by molecular biological factors, including factors related to the induction of apoptosis and the activity of proliferation. We analyzed immunohistochemically the expression of p53, Bcl-2, and Ki-67 in various types of cancers and assessed the correlation between this expression and chemosensitivity. Moreover, we investigated whether the expression of these factors could be a useful predictor for the clinical response to chemotherapy. Study subjects comprised 63 preoperative patients with untreated malignant tumors (9 with esophageal cancer, 12 with stomach cancer, 12 with colon cancer, 16 with liver cancer, and 14 with breast cancer). Immunohistochemical staining (the labeled streptavidin biotin technique: LSAB method) was used to assess expression of p53 protein, Bcl-2 protein, and Ki-67. A chemosensitivity test was carried out with the histoculture drug response assay method using four drugs: mitomycin C, 5 fluorouracil, doxorubicin hydrochloride (ADM), and cisplatin (CDDP). Immunohistochemical studies for p53 were found to be useful for predicting chemosensitivity. PMID- 15901939 TI - Role of impaired insulin secretion and insulin resistance in the pathogenesis of type 2 diabetes mellitus. AB - Type 2 diabetes mellitus (T2DM) is a heterogeneous disorder caused by a combination of genetic and acquired abnormalities that affect insulin sensitivity and insulin secretion. Currently available data suggest that insulin resistance is the acquired defect largely secondary to unhealthy lifestyles and that the major genetic factor is impaired insulin secretion. The latter is the result of both reduced beta-cell mass and functional abnormalities makes the beta-cell unable to compensate for increased insulin requirements caused by insulin resistance. Targeting both insulin resistance and impaired insulin secretion is therefore appropriate to prevent T2DM and to improve glycemic control in those with the disorder. PMID- 15901940 TI - Update on stroke prevention. AB - It is now well recognized that stroke-prone individual scan be identified in the majority of instances and that effective preventive therapy is available. Cumulative risk factors enhance the potential for stroke.The challenge is to effectively intervene to substantially reduce stroke risk. PMID- 15901941 TI - Prostate cancer screening. AB - Prostate cancer is the leading noncutaneous cancer in men of the Western world. Because of its prevalence and ability to cause morbidity and mortality,prostate cancer screening continues to be an important area of focus in health care. This article covers the sensitivity and specificity of prostate-specific antigen and current techniques used to improve the test's validity, the importance of detecting clinically important cancers with screening, as well as the downward stage migration, decreased disease-specific mortality, and decreased metastases rate seen inpatients screened and treated for prostate cancer. PMID- 15901942 TI - The management of large hemispheric cerebral infarcts. AB - Large hemispheric cerebral infarcts have significant morbidity and mortality. Our understanding of this pathophysiological process involved with secondary neurological deterioration in large hemispheric infarctions has increased in the past few decades. New experimental strategies designed to improve outcome are reviewed. PMID- 15901943 TI - Diagnosis and treatment of common sexually transmitted diseases. AB - Sexually transmitted diseases (STDs) are common and often are asymptomatic. This article reviews the presentation and treatment recommendations for some of the most common symptomatic STDs, as well as reviews recent advances in diagnostic methods that may impact patient care. PMID- 15901944 TI - Photodynamic therapy for Barrett's esophagus with high-grade dysplasia. AB - This article describes advances in photodynamic therapy for patients with Barrett's esophagus and high-grade dysplasia-an important, minimally invasive treatment option proven to safely and durably ablate Barrett's dysplasia and prevent carcinoma while preserving the gastroesophageal junction. PMID- 15901945 TI - Rosacea: the battle goes on. AB - Rosacea presents an enigma to patients and physicians alike. Although new insights and a plethora of therapies provide hope, the underlying etiology remains unknown. This assures a certain amount of frustration as available treatments temporize rather than cure the disease. This article examines the current state of knowledge regarding this fascinating entity. PMID- 15901946 TI - Comprehensive imaging of coronary artery disease: impact on contemporary treatment approaches. AB - Coronary artery disease remains a major cause of mortality. Together with novel therapeutic and preventive approaches, important advances of coronary imaging are currently emerging. This article describes the status of modern coronary imaging and outlines expected future developments. PMID- 15901947 TI - Time-domain scanning optical mammography: I. Recording and assessment of mammograms of 154 patients. AB - Using a triple wavelength (670 nm, 785 nm, 843/884 nm) scanning laser-pulse mammograph we recorded craniocaudal and mediolateral projection optical mammograms of 154 patients, suspected of having breast cancer. From distributions of times of flight of photons recorded at typically 1000-2000 scan positions, optical mammograms were derived displaying (inverse) photon counts in selected time windows, absorption and reduced scattering coefficients or total haemoglobin concentration and blood oxygen saturation. Optical mammograms were analysed by comparing them with x-ray and MR mammograms, including results of histopathology, attributing a subjective visibility score to each tumour assessed. Out of 102 histologically confirmed tumours, 72 tumours were detected retrospectively in both optical projection mammograms, in addition 20 cases in one projection only, whereas 10 tumours were not detectable in any projection. Tumour contrast and contrast-to-noise ratios of mammograms of the same breast, but derived from measured DTOFs by various methods were quantitatively compared. On average, inverse photon counts in selected time windows, including total photon counts, provide highest tumour contrast and contrast-to-noise ratios. Based on the results of the present study we developed a multi-wavelength, multi-projection scanning time-domain optical mammograph with improved spectral and spatial (angular) sampling, that allows us to record entire mammograms simultaneously at various offsets between the transmitting fibre and receiving fibre bundle and provides first results for illustration. PMID- 15901948 TI - Time-domain scanning optical mammography: II. Optical properties and tissue parameters of 87 carcinomas. AB - Within a clinical trial on scanning time-domain optical mammography reported on in a companion publication (part I), craniocaudal and mediolateral projection optical mammograms were recorded from 154 patients, suspected of having breast cancer. Here we report on in vivo optical properties of the subset of 87 histologically validated carcinomas which were visible in optical mammograms recorded at two or three near-infrared wavelengths. Tumour absorption and reduced scattering coefficients were derived from distributions of times of flight of photons recorded at the tumour site employing the model of diffraction of photon density waves by a spherical inhomogeneity, located in an otherwise homogeneous tissue slab. Effective tumour radii, taken from pathology, and tumour location along the compression direction, deduced from off-axis optical scans of the tumour region, were included in the analysis as prior knowledge, if available. On average, tumour absorption coefficients exceeded those of surrounding healthy breast tissue by a factor of about 2.5 (670 nm), whereas tumour reduced scattering coefficients were larger by about 20% (670 nm). From absorption coefficients at 670 nm and 785 nm total haemoglobin concentration and blood oxygen saturation were deduced for tumours and surrounding healthy breast tissue. Apart from a few outliers total haemoglobin concentration was observed to be systematically larger in tumours compared to healthy breast tissue. In contrast, blood oxygen saturation was found to be a poor discriminator for tumours and healthy breast tissue; both median values of blood oxygen saturation are the same within their statistical uncertainties. However, the ratio of total haemoglobin concentration over blood oxygen saturation further improves discrimination between tumours and healthy breast tissue. For 29 tumours detected in optical mammograms recorded at three wavelengths (670 nm, 785 nm, 843 nm or 884 nm), scatter power was derived from transport scattering coefficients. Scatter power of tumours tends to be larger than that of surrounding healthy breast tissue, yet the 95% confidence intervals of both medians overlap. PMID- 15901949 TI - Time-resolved optical mammography between 637 and 985 nm: clinical study on the detection and identification of breast lesions. AB - The first time-resolved optical mammograph operating beyond 900 nm was tested in a retrospective clinical study involving 194 patients with malignant and benign lesions, to investigate the diagnostic potential for the detection and characterization of breast lesions. For the first part of the study (101 patients with 114 lesions), the system was operated at 683, 785, 913 and 975 nm. Subsequently, to improve the spectral content of optical images, the number of wavelengths was increased (up to 7) and the spectral range was extended (637-985 nm). Late gated intensity and scattering images provide sensitivity to tissue composition (oxy- and deoxyhaemoglobin, water and lipids) and physiology (total haemoglobin content and oxygen saturation), as well as to structural changes. Tumours are typically identified because of the strong blood absorption at short wavelengths (637-685 nm), while cysts are characterized by low scattering, leading to a detection rate of approximately 80% for both lesion types, when detection is required in both cranio-caudal and oblique views. The detection rate for other benign lesions, such as fibroadenomas, is presently much lower (<40%). The effectiveness of the technique in localizing and identifying different lesion types was analysed as a function of various parameters (lesion size, compressed breast thickness, age, body mass index, breast parenchymal pattern). The possibility that physiologic changes due to the development of a malignant lesion could affect the entire breast was investigated. The capacity to assess the density of breast based on the average scattering properties was also tested. PMID- 15901950 TI - Characterization of female breast lesions from multi-wavelength time-resolved optical mammography. AB - Characterization of both malignant and benign lesions in the female breast is presented as the result of a clinical study that involved more than 190 subjects in the framework of the OPTIMAMM European project. All the subjects underwent optical mammography, by means of a multi-wavelength time-resolved mammograph, in the range 637-985 nm. Optical images were processed by applying a perturbation model, relying on a nonlinear approximation of time-resolved transmittance curves in the presence of an inclusion, with the aim of estimating the major tissue constituents (i.e. oxy- and deoxy-haemoglobin, lipid and water) and structural parameters (linked to dimension and density of the scatterer centres) for both the lesion area and the surrounding tissue. The critical factors for the application of the perturbation model on in vivo data are also discussed. Forty six malignant and 68 benign lesions were analysed. A subset of 32 cancers, 40 cysts and 14 fibroadenomas were found reliable for the perturbation analysis. For cancers, we show a higher blood content with respect to the surrounding tissue, while cysts are characterized by a lower concentration of scattering centres with respect to the surrounding tissue. For fibroadenomas, the low number of cases does not allow any definite conclusions. PMID- 15901951 TI - Optical tomography of the breast using a multi-channel time-resolved imager. AB - A time-resolved optical tomography system has been used to generate cross sectional images of the human breast. Images are reconstructed using an iterative, nonlinear algorithm and measurements of mean photon flight time relative to those acquired on a homogeneous reference phantom. Thirty-eight studies have been performed on three healthy volunteers and 21 patients with a variety of breast lesions including cancer. We have successfully detected 17 out of 19 lesions, and shown that optical images of the healthy breast of the same volunteer display a heterogeneity which is repeatable over a period of months. However, results also indicate that the lack of accurate quantitation of optical parameters and limited morphological information limits the ability to characterize different types of lesions and distinguish benign from malignant tissues. Drawbacks of our current methodology and plans for overcoming them are discussed. PMID- 15901952 TI - Reconstruction of optical properties of phantom and breast lesion in vivo from paraxial scanning data. AB - We report on the reconstruction of absorption and reduced scattering coefficients of breast tissue in vivo of a patient with mastopathic disease. Distributions of times of flight of photons through the compressed breast were recorded by paraxial scanning. From data measured at four different source-detector offsets optical properties were reconstructed within the linear Rytov approximation by a fast inverse Fourier space method. Low-pass filtering in Fourier space was employed to remove excessive noise from high spatial frequency components and to reduce the computational efforts by a factor of 3, typically. The mammograms displaying reconstructed absorption and reduced scattering coefficients were compared with projection mammograms either obtained by time-window analysis of experimental data or based on average absorption and reduced scattering coefficients which were derived from measured temporal point spread functions within a simple homogeneous model. All inhomogeneities which were visible in the projection mammograms and which could be associated with specific breast tissue compartments could be correlated with inhomogeneities in the reconstructed absorption coefficient. In particular, the mastopathic disease was detected in the reconstructed absorption mammogram. In order to assess reliability of optical properties reconstructed from data obtained by paraxial scanning, corresponding phantom experiments and reconstructions of phantom optical properties were carried out. Because of the limited angular range sampled by the in vivo and phantom measurements, considerable blurring of the absorption coefficient occurs along the compression direction, compromising longitudinal resolution. PMID- 15901953 TI - The Twente Photoacoustic Mammoscope: system overview and performance. AB - We present PAM, the Photoacoustic Mammoscope developed at the University of Twente, intended for initial retrospective clinical studies on subjects with breast tumours. A parallel plate geometry has been adopted and the breast will be gently compressed between a glass plate and a flat ultrasound detector matrix. Pulsed light (5 ns) from an Nd:YAG laser will impinge the breast through the glass plate in regions of interest; an appropriate number of the 590 elements of the detector matrix will be activated in succession to record photoacoustic signals. Three-dimensional image reconstruction employs a delay-and-sum beamforming algorithm. We discuss various instrumental aspects and the proposed imaging protocol. Performance studies of the ultrasound detector are presented in terms of sensitivity, frequency response and resolution. Details of the patient instrument interface are provided. Finally some imaging results on well characterized breast tissue phantoms with embedded tumour simulating inserts are shown. PMID- 15901954 TI - Characterization of normal breast tissue heterogeneity using time-resolved near infrared spectroscopy. AB - In recent years, extensive efforts have been made in developing near-infrared optical techniques to be used in detection and diagnosis of breast cancer. Variations in optical properties of normal breast tissue set limits to the performance of such techniques and must therefore be thoroughly examined. In this paper, we present intra- and intersubject as well as contralateral variations of optical and physiological properties in breast tissue as measured by using four wavelength time-resolved spectroscopy (at 660, 786, 916 and 974 nm). In total, 36 volunteers were examined at five regions at each breast. Optical properties (absorption, mu(a), and reduced scattering, mu'(s)) are derived by employing diffusion theory. The use of four wavelengths enables determination of main tissue chromophores (haemoglobin, water and lipids) as well as haemoglobin oxygenation. Variations in all evaluated properties seen over the entire breast are approximately twice those for small-scale heterogeneity (millimetre scale). Intrasubject variations in optical properties are almost in all cases below 20% for mu'(s), and 40% for mu(a). Overall variations in water, lipid and haemoglobin concentrations are all in the order of 20%. Oxygenation is the least variable of the quantities evaluated, overall intrasubject variations being 6% on average. Extracted physiological properties confirm differences between pre- and post menopausal breast tissue. Results do not indicate systematic differences between left and right breasts. PMID- 15901955 TI - Optical biopsy of breast tissue using differential path-length spectroscopy. AB - Differential path-length spectroscopy (DPS) was used to determine the local optical properties of breast tissue in vivo. DPS measurements were made on healthy and malignant breast tissue using a fibre-optic needle probe, and were correlated to the histological outcome of core-needle biopsies taken from the same location as the measurements. DPS yields information on the local tissue blood content, the local blood oxygenation, the average micro-vessel diameter, the beta-carotene concentration and the scatter slope. Our data show that malignant breast tissue is characterized by a significant decrease in tissue oxygenation and a higher blood content compared to normal breast tissue. PMID- 15901956 TI - Optical characterization of thin female breast biopsies based on the reduced scattering coefficient. AB - One of the main goals in optical characterization of biopsies is to discern between tissue types. Usually, the theory used for deriving the optical properties of such highly scattering media is based on the diffusion approximation. However, biopsies are usually small in size compared to the transport mean free path and thus cannot be treated with standard diffusion theory. To account for this, an improved theory was developed, by the authors, that can correctly describe light propagation in small geometries (Garofalakis et al 2004 J. Opt. A: Pure Appl. Opt. 6 725-35). The theory's limit was validated by both Monte Carlo simulations and experiments performed on tissue-like phantoms, and was found to be two transport mean free paths. With the aid of this theory, we have characterized 59 samples of breast tissue including cancerous samples by retrieving their reduced scattering coefficients from time-resolved transmission data. The mean values for the reduced scattering coefficients of the normal and the tumour tissue were measured to be 9.7 +/- 2.2 cm(-1) and 10.8 +/- 1.8 cm(-1), respectively. The correlation with age was also investigated. PMID- 15901957 TI - Characterization of Photofrin photobleaching for singlet oxygen dose estimation during photodynamic therapy of MLL cells in vitro. AB - A singlet oxygen dose model is developed for PDT with Photofrin. The model is based on photosensitizer photobleaching kinetics, and incorporates both singlet oxygen and non-singlet oxygen mediated bleaching mechanisms. To test our model, in vitro experiments were performed in which MatLyLu (MLL) cells were incubated in Photofrin and then irradiated with 532 nm light. Photofrin fluorescence was monitored during treatment and, at selected fluence levels, cell viability was determined using a colony formation assay. Cell survival correlated well to calculated singlet oxygen dose, independent of initial Photofrin concentration or oxygenation. About 2 x 10(8) molecules of singlet oxygen per cell were required to reduce the surviving fraction by 1/e. Analysis of the photobleaching kinetics suggests that the lifetime of singlet oxygen in cells is 0.048 +/- 0.005 micros. The generation of fluorescent photoproducts was not a result of singlet oxygen reactions exclusively, and therefore did not yield additional information to aid in quantifying singlet oxygen dose. PMID- 15901958 TI - A comparison of the performance of modern screen-film and digital mammography systems. AB - This work compares the detector performance and image quality of the new Kodak Min-R EV mammography screen-film system with the Fuji CR Profect detector and with other current mammography screen-film systems from Agfa, Fuji and Kodak. Basic image quality parameters (MTF, NPS, NEQ and DQE) were evaluated for a 28 kV Mo/Mo (HVL = 0.646 mm Al) beam using different mAs exposure settings. Compared with other screen-film systems, the new Kodak Min-R EV detector has the highest contrast and a low intrinsic noise level, giving better NEQ and DQE results, especially at high optical density. Thus, the properties of the new mammography film approach those of a fine mammography detector, especially at low frequency range. Screen-film systems provide the best resolution. The presampling MTF of the digital detector has a value of 15% at the Nyquist frequency and, due to the spread size of the laser beam, the use of a smaller pixel size would not permit a significant improvement of the detector resolution. The dual collection reading technology increases significantly the low frequency DQE of the Fuji CR system that can at present compete with the most efficient mammography screen-film systems. PMID- 15901959 TI - Progress towards in vitro quantitative imaging of human femur using compound quantitative ultrasonic tomography. AB - The objective of this study is to make cross-sectional ultrasonic quantitative tomography of the diaphysis of long bones. Ultrasonic propagation in bones is affected by the severe mismatch between the acoustic properties of this biological solid and those of the surrounding soft medium, namely, the soft tissues in vivo or water in vitro. Bone imaging is then a nonlinear inverse scattering problem. In this paper, we showed that in vitro quantitative images of sound velocities in a human femur cross section could be reconstructed by combining ultrasonic reflection tomography (URT), which provides images of the macroscopic structure of the bone, and ultrasonic transmission tomography (UTT), which provides quantitative images of the sound velocity. For the shape, we developed an image-processing tool to extract the external and internal boundaries and cortical thickness measurements. For velocity mapping, we used a wavelet analysis tool adapted to ultrasound, which allowed us to detect precisely the time of flight from the transmitted signals. A brief review of the ultrasonic tomography that we developed using correction algorithms of the wavepaths and compensation procedures are presented. Also shown are the first results of our analyses on models and specimens of long bone using our new iterative quantitative protocol. PMID- 15901960 TI - A complete distortion correction for MR images: II. Rectification of static-field inhomogeneities by similarity-based profile mapping. AB - Radiotherapy treatment planning relies on the use of geometrically correct images. This paper presents a fully automatic tool for correcting MR images for the effects of B(0) inhomogeneities. The post-processing method is based on the gradient-reversal technique of Chang and Fitzpatrick (1992 IEEE Trans. Med. Imaging 11 319-29) which combines two identical images acquired with a forward- and a reversed read gradient. This paper demonstrates how maximization of mutual information for registration of forward and reverse read gradient images allows the elimination of user interaction for the correction. Image quality is preserved to a degree not reported previously. PMID- 15901961 TI - Analytical solutions of electric potential and impedance for a multilayered spherical volume conductor excited by time-harmonic electric current source: application in brain EIT. AB - A model of a multilayered spherical volume conductor with four electrodes is built. In this model, a time-harmonic electric current is injected into the sphere through a pair of drive electrodes, and electric potential is measured by the other pair of measurement electrodes. By solving the boundary value problem of the electromagnetic field, the analytical solutions of electric potential and impedance in the whole conduction region are derived. The theoretical values of electric potential on the surface of the sphere are in good accordance with the experimental results. The analytical solutions are then applied to the simulation of the forward problem of brain electrical impedance tomography (EIT). The results show that, for a real human head, the imaginary part of the electric potential is not small enough to be ignored at above 20 kHz, and there exists an approximate linear relationship between the real and imaginary parts of the electric potential when the electromagnetic parameters of the innermost layer keep unchanged. Increase in the conductivity of the innermost layer leads to a decrease of the magnitude of both real and imaginary parts of the electric potential on the scalp. However, the increase of permittivity makes the magnitude of the imaginary part of the electric potential increase while that of the real part decreases, and vice versa. PMID- 15901962 TI - Estimation of electrical conductivity distribution within the human head from magnetic flux density measurement. AB - We have developed a new algorithm for magnetic resonance electrical impedance tomography (MREIT), which uses only one component of the magnetic flux density to reconstruct the electrical conductivity distribution within the body. The radial basis function (RBF) network and simplex method are used in the present approach to estimate the conductivity distribution by minimizing the errors between the 'measured' and model-predicted magnetic flux densities. Computer simulations were conducted in a realistic-geometry head model to test the feasibility of the proposed approach. Single-variable and three-variable simulations were performed to estimate the brain-skull conductivity ratio and the conductivity values of the brain, skull and scalp layers. When SNR = 15 for magnetic flux density measurements with the target skull-to-brain conductivity ratio being 1/15, the relative error (RE) between the target and estimated conductivity was 0.0737 +/- 0.0746 in the single-variable simulations. In the three-variable simulations, the RE was 0.1676 +/- 0.0317. Effects of electrode position uncertainty were also assessed by computer simulations. The present promising results suggest the feasibility of estimating important conductivity values within the head from noninvasive magnetic flux density measurements. PMID- 15901963 TI - Interaction of mobile phones with superficial passive metallic implants. AB - The dosimetry of exposure to radiofrequency (RF) electromagnetic (EM) fields of mobile phones is generally based on the specific absorption rate (SAR, W kg(-1)), which is the electromagnetic energy absorbed in the tissues per unit mass and time. In this study, numerical methods and modelling were used to estimate the effect of a passive, metallic (conducting) superficial implant on a mobile phone EM field and especially its absorption in tissues in the near field. Two basic implant models were studied: metallic pins and rings in the surface layers of the human body near the mobile phone. The aim was to find out 'the worst case scenario' with respect to energy absorption by varying different parameters such as implant location, orientation, size and adjacent tissues. Modelling and electromagnetic field calculations were carried out using commercial SEMCAD software based on the FDTD (finite difference time domain) method. The mobile phone was a 900 MHz or 1800 MHz generic phone with a quarter wave monopole antenna. A cylindrical tissue phantom models different curved sections of the human body such as limbs or a head. All the parameters studied (implant size, orientation, location, adjacent tissues and signal frequency) had a major effect on the SAR distribution and in certain cases high local EM fields arose near the implant. The SAR values increased most when the implant was on the skin and had a resonance length or diameter, i.e. about a third of the wavelength in tissues. The local peak SAR values increased even by a factor of 400-700 due to a pin or a ring. These highest values were reached in a limited volume close to the implant surface in almost all the studied cases. In contrast, without the implant the highest SAR values were generally reached on the skin surface. Mass averaged SAR(1 g) and SAR(10 g) values increased due to the implant even by a factor of 3 and 2, respectively. However, at typical power levels of mobile phones the enhancement is unlikely to be problematic. PMID- 15901964 TI - Theoretical and experimental validation of treatment planning for narrow MLC defined photon fields. AB - In intensity modulated radiotherapy (IMRT), the use of small fields where electronic equilibrium does not exist is becoming more common and presents difficulties for both the measurement and calculation of dose to such fields. Pinnacle(3) (Version 6.2b) allows the user to specify a total minimum open area for each IMRT segment, which can result in sub-segments with widths of only a few millimetres. The dose for 6 MV narrow MLC defined fields between 0.1 and 3 cm in width was investigated using Kodak extended dose range film (EDR2), ionization chamber and MOSFET dosimeters and BEAMnrc Monte Carlo calculations, and these results were used to determine the accuracy of Pinnacle(3) dose calculation for narrow MLC segments. The incident fluences calculated by Pinnacle(3) and BEAMnrc were also compared. Results show that if a fluence and dose grid resolution of 0.1 cm is used, Pinnacle(3) dose agrees with the EDR2 and BEAMnrc to within 5% for field widths between 0.5 and 3.0 cm. However, Pinnacle(3) will underestimate the dose by up to 45% for the 0.1 and 0.3 cm wide fields. It is shown that the source size in the Pinnacle(3) beam model and both the fluence and dose grid resolutions have a significant effect on the accuracy of dose calculation for field widths of 1.0 cm and less. For single segment fields, Pinnacle(3) agrees with EDR2 and BEAMnrc to within 0.1 cm at the field edges and underestimates the penumbra width by up to 0.08 cm. Results for multiple segment fields showed that an MLC transmission of 1.7% and a 0.06 cm inward shift of MLCs prior to beam delivery gave the closest agreement between Pinnacle(3) and measurement. The multiple segment fields also revealed a pattern of low dose troughs of up to 7% in the Pinnacle(3) dose profiles. PMID- 15901965 TI - Demonstration of a forward iterative method to reconstruct brachytherapy seed configurations from x-ray projections. AB - By monitoring brachytherapy seed placement and determining the actual configuration of the seeds in vivo, one can optimize the treatment plan during the process of implantation. Two or more radiographic images from different viewpoints can in principle allow one to reconstruct the configuration of implanted seeds uniquely. However, the reconstruction problem is complicated by several factors: (1) the seeds can overlap and cluster in the images; (2) the images can have distortion that varies with viewpoint when a C-arm fluoroscope is used; (3) there can be uncertainty in the imaging viewpoints; (4) the angular separation of the imaging viewpoints can be small owing to physical space constraints; (5) there can be inconsistency in the number of seeds detected in the images; and (6) the patient can move while being imaged. We propose and conceptually demonstrate a novel reconstruction method that handles all of these complications and uncertainties in a unified process. The method represents the three-dimensional seed and camera configurations as parametrized models that are adjusted iteratively to conform to the observed radiographic images. The morphed model seed configuration that best reproduces the appearance of the seeds in the radiographs is the best estimate of the actual seed configuration. All of the information needed to establish both the seed configuration and the camera model is derived from the seed images without resort to external calibration fixtures. Furthermore, by comparing overall image content rather than individual seed coordinates, the process avoids the need to establish correspondence between seed identities in the several images. The method has been shown to work robustly in simulation tests that simultaneously allow for unknown individual seed positions, uncertainties in the imaging viewpoints and variable image distortion. PMID- 15901966 TI - Effects of a novel cyclohexane dicarboximide derivative, ST-6, on reperfusion induced arrhythmia in rats. AB - The present study was designed to determine whether a novel cyclohexane dicarboximide derivative, ST-6, 2-[4-[4-(chlorophenyl)-4-hydroxy-1 piperidinyl]butyl]hexahydro-1H-isoindol-1,3(2H)-dione, prevents reperfusion induced ventricular arrhythmias. Pentobarbital-anesthetized rats were subjected to left coronary artery occlusion for 4 min followed by 4-min reperfusion, and the incidence of their ventricular arrhythmias was examined. The coronary occlusion of control rats induced ventricular tachycardia and fibrillation, eventually leading to sudden death. The intravenous injection of 0.1 to 2 mg/kg ST-6 prior to the occlusion resulted in a dose-dependent suppression of the ventricular arrhythmias. The suppression of ventricular fibrillation was also observed on the intraperitoneal and intradoudenal administration of 2 to 10 mg/kg ST-6 15 min prior to coronary occlusion. Antiarrhythmic effects of this agent (0.5 mg/kg per min) were compared with those of other antiarrhythmic agents including lidocaine (0.1 mg/kg per min), sematilide (0.3 mg/kg per min), and diltiazem (0.5 mg/kg per min) by administrating the agents from 1 min after the coronary occlusion to the end of 4-min reperfusion. Antiarrhythmic effects of ST 6 were similar in degree to those of lidocaine and diltiazem, whereas no significant prevention by sematilide was seen. The results suggest that ST-6 may be capable of suppressing reperfusion-induced arrhythmias following oral or intravenous administration. PMID- 15901967 TI - [Development of nephrology in Lithuania]. AB - This article reviews the development of nephrology in the different regions of Lithuania. History of Lithuanian nephrology started in Vilnius in 1969 when the first hemodialysis was performed in the oldest hospital of Lithuania. In 1969 the clinics of Kaunas Medical University started hemodialysis service, in 1977 Panevezys hospital, in 1991-Siauliai hospital and in 1993-Klaipeda hospital. In 1965 Assoc. Prof. S. Markovicius began to give a course in nephrology at the Faculty of Medicine of Vilnius University. In 1967 the first department of nephrology in Lithuania was opened at Vilnius City Hospital. PMID- 15901968 TI - Left ventricular geometry in children with chronic renal failure. AB - The aim of the study was to assess left ventricular (LV) geometry in children with chronic renal insufficiency (CRI), and its relationship with glomerular filtration rate (GFR) and preexisting renal disease. Echocardiography was performed on 56 non-dialysed CRI patients and 56 controls. CRI patients had bigger interventricular septum thickness (0.77+/-0.17 vs. 0.67+/-0.12 cm, p=0.002), LV posterior wall thickness (0.79+/-0.14 vs. 0.71+/-0.14 cm, p<0.006), LV mass index (LVMI) (40.7+/-12.2 vs. 31.7+/-6.3 g/m2.7, p<0.0001; 86.4+/-24.1 vs. 69.1+/-13.9 g/m2, p<0.0001), and relative wall thickness (0.38+/-0.05 vs. 0.34+/-0.04, p<0.0001) in comparison with controls. Twenty (36%) of CRI patients had LV hypertrophy (LVH). Thirteen patients (23%) had eccentric LVH, 7 (13%) concentric LVH), and 9 (16%) of patients-concentric LV remodeling. No significant difference was found between LV parameters in patient groups with different GFR. Patients with acquired renal diseases and hereditary nephropathies had significantly higher LVMI than patients with congenital renal abnormalities. Our results indicate that changes of LV geometry are present in children with mild, moderate and predialysis CRI. These findings support the concept of cardiovascular disease risk for patients with different stages of CRI. PMID- 15901969 TI - [Hemolytic-uremic syndrome in Lithuania over the period 1990-2004]. AB - More than 80 cases of the hemolytic-uremic syndrome were registered in Lithuania over the period 1990-2004. The incidence of this syndrome fluctuated annually; higher incidence rates were observed in 1990-1993, 1998-1999 and 2003-2004, however, there was no overall increase in morbidity over the 15-year period. Most cases occurred in the first two years of life; besides, girls suffered slightly more often than boys. At least in 80% of cases the illness was typical, i.e. was associated with diarrhea. In general, the data correspond to the findings of other scientists. PMID- 15901970 TI - [Dysfunction of lower urinary tract in 5-18 years old children with recurrent urinary tract infections]. AB - The purpose of this article was to evaluate function of lower urinary tract for 5 18 years old children with recurrent urinary tract infections and to compare urodynamic changes in two groups: with recurrent lower urinary tract infections and with recurrent upper urinary tract infections. 35 urodynamic studies in 5-18 years old children (3 boys and 32 girls) with recurrent urinary tract infection were performed at the Clinic of Children's Diseases of Kaunas University of Medicine in 2004. 21 of these children had recurrent lower urinary tract infections and 14 recurrent upper urinary tract infections. Changes in urodynamics were present in 91.4% of children: in all children with recurrent lower urinary tract infections and in 11 children with upper recurrent urinary tract infections (78.6%). Detrusor instability was found in 20 (57.1%) children: in 14 (66,7%) with recurrent lower urinary tract infections and in 6 (42.9%) with recurrent upper urinary tract infections. In 32 (85%) children detrusor instability was accompanied by changes in bladder volume. Changes of bladder volume were present for 26 (74.3%) children with recurrent urinary tract infections. Detrusor after contraction was diagnosed in 57.1% of children with recurrent lower urinary tract infections and in 28.6% with recurrent upper urinary tract infections. For children with recurrent urinary tract infections attention for urinary tract dysfunction must be paid. PMID- 15901971 TI - [Etiology and outcomes of acute renal failure in childhood]. AB - The aim of the research was to determine causes of acute renal failure in children, their outcome and to define risk factors associated with mortality. 75 children with acute renal failure, who were treated at the Clinic of Children's Diseases of Kaunas University of Medicine between 1998-2003 years, were included in the study. The age range of patients was 1 month to 16 years. They were divided into two groups. Acute renal failure was diagnosed in 42 (56%) patients (the first study group) and in 33 (44%) patients acute renal failure was together with multiple organ failure (the second study group). In the first study group 69% of cases of acute renal failure were found to be due to renal diseases and in the second study group 97% were because of extrarenal diseases. Sepsis was the most frequent cause of acute renal failure in the second group (p<0.02). Dialysis was made for 28% patients. Hypertension was diagnosed more often in the first patients group (p<0.05). Hypertension persisted in 9 (36%) patients after recovery. Chronic renal failure developed in two patients. 28 (37.3%) patients of the original study group died. Mortality rate for children with multiple organ failure was higher than for the children, who had renal insufficiency only (78.8% vs 4.8%; p<0.001). Mortality rate of infants in the first study group was higher than for children of the same age in the second group (p<0.001). Mortality rate for children, who had oliguria or anuria, was higher in the second group, too (p<0.001). PMID- 15901972 TI - Comparative study of prednisolone alone and prednisolone plus fusidic acid in the treatment of children with steroid-responsive nephrotic syndrome. AB - BACKGROUND: There are several reports in the literature indicating that fusidic acid owns functions similar to those of cyclosporin. As cyclosporin has effectively been used in frequently relapsing steroid-responsive nephrotic syndrome we carried out this study to determine whether fusidic acid used along with prednisolone diminishes rate of steroid-responsive nephrotic syndrome relapses in children. PATIENTS AND METHODS: The patients were randomly allocated to receive either prednisolone alone or prednisolone plus fusidic acid for two months in standard doses. In the cases of relapses the treatment was changed to the comparative treatment method. Altogether 18 children (12 boys and 6 girls) aged 1.3 to 13.2 years entered the study. Thirteen of them were treated by either method on different occasions, four patients were treated with prednisolone only, and one child was treated with prednisolone plus fusidic acid only. Thus, there were 17 evaluable treatment courses with prednisolone alone and 14 courses with prednisolone plus fusidic acid. The patients were followed-up as long as the remission lasted. RESULTS: There was prompt and complete response under the influence of both treatment methods. However, relapses occurred in all the patients irrespective of the mode of treatment. Mean duration of remissions did not differ significantly between the study groups (17.8+/-20.4 weeks in the prednisolone group and 18.3+/-23.9 weeks in the prednisolone plus fusidic acid group; p>0.05). There were no statistically significant differences in laboratory parameters reflecting therapeutic efficacy in the comparative treatment groups, too. CONCLUSION: Thus, it was not revealed any remission-sustaining efficacy of fusidic acid used in standard doses for two months along with prednisolone in children with frequently relapsing steroid-responsive nephrotic syndrome. PMID- 15901973 TI - [Development of hemodialysis service in Lithuania during (1996-2003)]. AB - The aim of our study was to evaluate the changes of hemodialysis (HD) service and main demographic characteristics of HD patients in Lithuania during seven years period. From 1996 to 2003 we visited annually all HD centers in Lithuania and collected data about all HD patients. There was a sharp increase in the number of HD centers (from 17 to 37), HD stations (from 25 p. m. p. (per million population) to 87 p.m.p.; p<0.001), HD patients (from 60 p.m.p. to 264 p.m.p.; p<0.001) and incidence of new HD patients (from 54.3 p.m.p. to 92 p.m.p.; p<0.01). In 1996 all 17 HD centers in Lithuania were public. Private HD centers appeared in 1998 and reached 43.2% of all HD centers (n=16) in 2003. 44.8% of HD patients were dialyzed in private HD centers. The mean age of HD patients increased from 47.2+/-16.1 years in 1996 to 57.5+/-14.9 years in 2003 (p<0.001). HD population became older. The percentage of patients over 60 years old increased from 22.8% to 49.7% (p<0.001) and over 70 years old--from 54% to 21.9% (p<0.001). The main underlying disease of end-stage renal disease was chronic glomerulonephritis but its rate had decreased from 54.5% in 1996 to 26.5% in 2003 (p<0.001). During this period there was a statistically significant increase in the incidence of end-stage renal disease due to diabetics (from 7.1% to 18.0%; p<0.01), hypertensive nephropathy (from 3.1% to 9.4%; p<0.05), and chronic pyelonephritis (from 11.2% to 18.6%; p<0.01). In summary during the last seven years HD service in Lithuania expanded significantly, and rapid development of private HD was observed. The number of HD patients was rising continuously with predominance of diabetic, hypertensive and elderly patients. PMID- 15901974 TI - [Changes of control of disorders of calcium and phosphorus metabolism in Lithuanian hemodialysis centers 1996-2003]. AB - The aim of the study was to evaluate the changes of the rate of disorders of calcium and phosphorus metabolism and their control in patients on hemodialysis (HD) in Lithuania in 1996-2003. Every December during this period we visited all HD centers of Lithuania and collected data on calcium-phosphorus metabolism in HD patients. 51.8% of HD patients in 1999 and 44.6% in 2003 had hyperphosphatemia (>1.8 mmol/l) (p<0.05). The mean phosphate concentration was 1.82+/-0.56 mmol/l in 2003 (p<0.05, comparing with 1.95+/-0.72 mmol/l in 1999 and 1.9+/-0.72 mmol/l in 2001). 7.1% of HD patients had hypocalcemia in 2003 and 7.8% hypercalcemia. Serum parathyroid hormone level was investigated only in 27.3% of HD patients in 1999 and 84.8% in 2003 (p<0.05). Use of alfacalcidol significantly decreased from 77.5% in 1998 to 29.4% in 2003, when the evaluation of serum parathyroid hormone increased (r=-0.911, p=0.03). Serum parathyroid hormone level was not analyzed for 59.8% of patients who used alfacalcidol and 59.4% of them had hyperphosphatemia in 1999 (6.3% and 32.9% in 2003, respectively; p<0.05). 10.7% of these patients had hypercalcemia in 2003. In summary, the correction of disorders of calcium and phosphorus metabolism in HD patients was insufficient but ameliorative. Monitoring of serum parathyroid hormone increased significantly during 1997-2003. The percentage of the precarious use of alfacalcidol decreased significantly when the evaluation of serum parathyroid hormone level became regular. PMID- 15901975 TI - [Early risk factors for secondary hyperparathyroidism in hemodialysis patients]. AB - The aim of the study was to identify risk factors for secondary hyperparathyroidism at the start and during the first year of hemodialysis. Retrospective analysis of medical records of all patients with end-stage renal disease, dialyzed at the hemodialysis center of Kaunas University of Medicine Hospital on December 2004, was performed. Biochemical data at the start, during the first year and at the end of follow-up (December 2004) were analyzed. At the start of hemodialysis elevated level of parathyroid hormone (PTH) was observed in 46 of 69 patients (67.6%), at the end of the first year in 27 of 69 patients (39.1%). In 22 of 46 patients (47.8%), who started hemodialysis with elevated PTH levels, the level of PTH decreased to <22 pmol/l after 1 year. In comparison with patients who maintained elevated PTH levels, they showed lower levels of PTH at the start of hemodialysis. Levels of serum calcium and phosphate at the start of hemodialysis did not differ between groups. At the end of the first year patients in whom levels of PTH decreased had higher serum calcium concentration as compared with those who maintained levels of PTH > or =22 pmol/l. A multivariate analysis revealed that levels of PTH and alkaline phosphatase after one year of hemodialysis were associated with increased risk for secondary hyperparathyroidism on the follow up. CONCLUSIONS: Control of calcium-phosphate metabolism in the pre-dialysis period is not sufficient. Symptoms of secondary hyperparathyroidism were diagnosed in more than a half of patients starting hemodialysis. Levels of PTH and alkaline phosphatase at the end of the first year are early risk factors for secondary hyperparathyroidism on the follow-up. PMID- 15901976 TI - [Surgical treatment of secondary hyperparathyroidism (The first experience in Klaipeda region)]. AB - The aim of the study was to analyze criteria for selection of patients for parathyroidectomy, results of surgery and the need for such surgeries in patients undergoing dialysis in Klaipeda region. RESULTS: 201 chronic patients were hemodialyzed in Klaipeda region in January 2005. 4 of them underwent parathyroidectomy. Before surgery parathyroid hormone of all patients was greater than 1000 pg/ml; during sonoscopy enlarged parathyroid glands were identified; increased levels of alkaline phosphatase and bone lesions, hyperphosphatemia hardly corrected with drugs were detected; one patient had tissue calcification. After surgery clinical state of all the patients improved noticeably and amounts of parathyroid hormone, calcium and phosphorus in blood decreased. No postoperative complications were observed. After review of medical records of 201 chronic patients undergoing hemodialysis, it was discovered that 9.4 % of patients had secondary hyperparathyroidism resistant to treatment. CONCLUSIONS: The first parathyroidectomies in Klaipeda region were successful; previous disorders of metabolism of calcium, parathyroid hormone and phosphorus were corrected. On average 9.4% of patients on hemodialysis in Klaipeda region were recommended to undergo parathyroidectomy. PMID- 15901977 TI - [Hospitalizations and comorbidity of diabetic patients on hemodialysis]. AB - We analysed ambulatory case records of all patients dialysed in 7 hemodialysis centers of Kaunas region during the year 2002 (n=186). Separate analysis of 42 diabetic patients was performed and compared with non-diabetics. Mortality was higher in diabetic patients in comparison with non-diabetics (22.5% and 7.3%, respectively; p<0.01). The mean number of associated diseases was 3.0+/-1.2 per diabetic patient. Disability status was worse in diabetic patients (p<0.01). Hospitalization rate of diabetic patients was 1.5 hospitalizations per patient year at risk, length 27.3 days per patient-year at risk; duration of one hospital stay was 18 days. The most frequent causes of hospitalization: infections (33% of all hospitalizations), diabetes complications (20%) and cardiovascular pathology (18%). Due to infections diabetic patients on hemodialysis were treated in hospital at longest. In incident diabetic patients (n=16) late referral was observed in 50% of patients and hemodialysis was started through central venous catheter. Starting hemodialysis diabetic patients were treated for longer periods in hospital than non-diabetic patients (duration of one hospitalization 40.6+/ 24.4 and 21.3+/-10.4 days, respectively; p<0.001). PMID- 15901978 TI - [Influence of anemia on hospitalization and mortality in hemodialysis patients]. AB - The objective of our study was to evaluate the influence of anemia on hospitalization and mortality in hemodialysis patients of Kaunas region. We analyzed ambulatory case records of 148 patients dialyzed in all 7 hemodialysis centers of Kaunas region in November 2001. The study consisted of two parts: in the first part data on patient age, gender, primary cause of end-stage renal disease, hemoglobin concentration were collected in November 2001 and in the second part these patients were followed up for 12 months in order to evaluate rate and length of hospitalization, reasons for hospitalization and mortality. At the beginning of the study mean hemoglobin (Hb) value was 101.2+/-13.8 g/l and more than a half of the patients (59%) had hemoglobin value higher than 100 g/l. Further follow-up of the patients during the year 2002 revealed that hemoglobin level of the patients who died was lower from the patients who followed hemodialysis. Mean hemoglobin of dead and alive patients was 92.4+/-18.6 g/l and 102.81+/-12.48 g/l, respectively (p=0.02). Lower hemoglobin concentrations were associated with a 5% higher relative risk of mortality for every 1 g/l decrease in hemoglobin (p=0.027). Analysis showed that mean hemoglobin value was 104.2+/ 11.1 g/l for the patients who were not hospitalized and 99+/-15.1 g/l for the patients who were hospitalized during the year 2002 (p=0.02). Lower hemoglobin concentrations were associated with a 5% higher relative risk for hospitalization for every 1 g/l decrease in hemoglobin (p=0.027). Patients who had mean hemoglobin value lower than 100g/l were hospitalized more frequently (p=0.01) and for longer period of time (p=0.005) than the patients with hemoglobin higher than 100 g/l. Analysis of the reasons for hospitalizations revealed that every 1 g/l decrease in hemoglobin increases relative risk for hospitalization due infections by 1% (p=0.000). PMID- 15901979 TI - [Zinc and aluminum concentrations in blood of hemodialysis patients and its impact on the frequency of infections]. AB - A multicenter study was performed with the aim to investigate the blood concentrations of zinc (Zn) and aluminum (Al) in hemodialysis patients and to clarify the impact of these metals level on the frequency of infections and the impact of medications on metal concentration in blood of these patients. MATERIAL AND METHODS: 265 patients with end-stage renal disease from 7 dialysis centers were enrolled in this study. All patients underwent standard hemodialysis. Venous blood samples were collected from hemodialysis patients before hemodialysis sessions. Atomic absorption spectrophotometry was applied to measure blood levels of Zn and Al. RESULTS AND CONCLUSIONS: The hemodialysis patients with infectious complications had significantly higher mean blood Al than the patients without complications (33.1+/-38.9 and 24.5+/-45.9 microg/l, respectively; p<0.001) and lower plasma levels of Zn (821.9+/-389.5 and 905.1+/-346.6 microg/l, respectively; p<0.005). Infectious complications were associated with zinc deficiency (p<0.01). The HD patients who were taken CaCO3 had lower mean concentration of Zn (849.4+/-324.6 and 1022.1+/-507.4 microg/l, respectively; p<0.006). PMID- 15901980 TI - [Sleep disorders and quality of life in patients on hemodialysis]. AB - The objective of this study was to determine the rate of sleep disorders in patients on hemodialysis and to evaluate the association between quality of sleep and quality of life in these patients. METHODS: A total of 81 hemodialysis patients were enrolled in the study. Quality of sleep was assessed using the Pittsburgh Sleep Quality Index (PSQI): higher scores indicate worse sleep quality. Health-related quality of life was evaluated using the Medical Outcomes Study 36-item Short Form (SF-36) questionnaire. RESULTS: In the present study 54 dialysis patients (66.7%) were "poor sleepers" (PSQI>5). The SF-36 mental component summary (PCS) and physical component summary (MCS) correlated with the global PSQI score (PCS, r=-0.463, p<0.001; MCS, r=-0.426, p<0.001), age (PCS, r= 0.330, p=0.003; MCS, r=-0.381, p<0.001), hemoglobin (PCS, r=0.289, p=0.009; MCS, r=0.301, p=0.006), comorbidity (PCS, r=-0.286, p=0.01; MCS, r=-0.283, p=0.011). Dialysis patients with global PSQI< or =5 ("good sleepers") had higher SF-36 PCS and MCS scores (PCS, 51.15+/-17.2 vs. 34.72+/-16.58, p<0.001; MCS, 59.52+/-17.43 vs. 41.92+/-18.34, p<0.001) and higher hemoglobin levels (102.74+/-12.34 g/l vs. 95.67+/-10.57 g/l, p=0.009) compared with "poor sleepers" (PSQI>5). CONCLUSIONS: In the present study two-thirds of dialysis patients (66.7%) were "poor sleepers". Lower hemoglobin levels correlated with worse quality of sleep and quality of life. We hypothesize that correction of anemia may improve quality of life in patients on hemodialysis. Poor sleep is associated with lower quality of life in hemodialysis patients. PMID- 15901981 TI - [Early thrombosis of radiocephalic fistula in patients on chronic hemodialysis (results of Kaunas University of Medicine Hospital 2000-2003)]. AB - In 2000-2003 surgeons performed 616 surgeries on arterio-venous access formation, more than 30% of them were radiocephalic. In 36.9% of cases of radiocephalic fistula early thrombotic complications developed (during the first month after operation) and a new access creation was needed. Various factors that indicate early thrombotic complications in radiocephalic fistula were assessed. 146 case records were analyzed retrospectively and all patients were divided into two groups. The first group (n=94) consisted of the patients with normal function of radiocephalic fistula; the second group (n=52) was comprised of patients in whom thrombotic complications developed during the first month after surgery. Comparison of the groups showed that the main disease, patients age, gender and technique of operation (all surgeries were performed using microsurgical technique, but accesses were formed "end to end" or "end to side" using 7/0 or 8/0 sutures), did not influence the development of early thrombotic complications in radiocephalic fistula. The shorter time of maturation of new access was associated with more frequent early thrombotic complications: the mean time of new vascular access maturation was 15.45+/-2.56 days in the first group, and 12.5+/-2.42 days in the second group (p<0.05). A marked tendency of more frequent early thrombotic complications was noticed in patients older than 60 years and in patients with ischemic heart disease. PMID- 15901982 TI - [Factors influencing survival of hemodialysis patients: data from hemodialysis center of Kaunas University of Medicine Hospital 1994-2004]. AB - We analyzed data of 383 hemodialysis patients admitted to the hemodialysis center of Kaunas University of Medicine Hospital between 1 January 1994 and 31 December 2004. The aim of the study was to estimate their survival and identify it influencing factors. Demographic data (gender, birth date), cause of renal insufficiency, date of hemodialysis (HD) initiation, blood analyses at the start of HD (hemoglobin, C-reactive protein, serum albumin, creatinine, calcium, phosphate), how HD were started (through central venous catheter or permanent vascular access), time of the first nephrologist consultation before initiating of HD were recorded for each patient in a special form. The total survival rate was estimated using the Kaplan-Maier method. Mean survival of HD patients was only 21.93 months (95% confidence intervals (CI) 18.7-25.16 months). It was influenced by a high early mortality (17.23% of patients died within the first three months from the beginning of HD (36.5% of all dead patients)). The lowest survival rate was of those patients who started HD immediately after the first consultation with a nephrologist. Early referral to nephrologist, normal serum albumin and C-reactive protein concentrations had a positive impact on survival in hemodialysis patients. PMID- 15901983 TI - [The beginning of kidney transplantation in Kaunas (results of Kaunas University of Medicine Hospital 2000-2004)]. AB - During the last year 57 cadaver kidney transplantations were performed in Lithuania, 23 of them in Kaunas. The first transplantation of cadaver kidney at Kaunas University of Medicine Hospital was performed in May 2000. The purpose of the study was to analyze the results of kidney transplantations performed at Kaunas University of Medicine Hospital between May 2000 and December 2004 and to evaluate the most frequent early and late complications after transplantation and survival of the kidney transplant. During this period a total of 51 cadaver kidney transplantations were done. The mean age of patients was 38.37+/-12.45 years. 90.2% of cadaver kidneys were taken from the optimal kidney donors. The most common early complications after kidney transplantation were infections of the urinary tract (33.3%) and on the late period of kidney transplantation complications of the immunosuppressive treatment (40%) (cytomegalovirus infection, agranulocytosis). Acute rejections were in 13.7 proc. of patients. Kidney transplant survival rate at 1 and 3 years after cadaver kidney transplantation was 85%. Mortality of transplant patients was 2%. PMID- 15901984 TI - [The first experience with sirolimus (Rapamune) after kidney transplantation in Lithuania]. AB - Sirolimus is a new immunosuppressive agent. This study aimed to evaluate the efficiency of sirolimus in patients after renal transplantation and to compare graft function, the frequency of rejection episodes and complications with patients under cyclosporin A treatment. From May 2002 to January 2005 26 renal transplant patients were treated with sirolimus. 13 patients (group A) were treated with sirolimus before renal transplantation and 13 patients (group B) were converted to sirolimus in late period after transplantation because of chronic cyclosporin A nephrotoxicity, chronic graft nephropathy and due to intolerance of cyclosporin A (mean time after transplantation: 18 months). Sirolimus was started as a loading dose 5-6 mg per day and reduced to 2-3 mg per day. Mean sirolimus blood concentration was 8.19+/-6.7 ng/ml. Results were compared according to age, gender, the number of HLA matches, plasma renin activity levels, etc., with 52 patients (control (C) group) under cyclosporin A, mycophenolate mofetil and steroids treatment. During 3 months, the acute rejections were in 30.8% of patients (4/13) and 65.4% of patients (17/26) for group A and group B, respectively (chi2=6.568, p<0.05). Renal function at 12 months: mean serum creatinine was 165.5+/-29 micromol/l vs. 214.2+/-67.9 micromol/l, urea 9.6+/-2.6 mmol/l vs. 13.9+/-9.3 mmol/l. There were no differences in platelet counts between groups, but serum cholesterol value was higher in the patients of group A (8.11+/-0.9 mmol/l vs. 6.54+/-1.4 mmol/l), blood pressure (140+/-13/87+/-14 mmHg vs. 150+/-15/85+/-12 mmHg). Patients were treated for different infections, cytomegalovirus infection and sepsis (28.6% (6/21) vs. 45.2% (19/52) for group A and group B, respectively). Our results have shown that sirolimus in combination with mycophenolate mofetil and steroids is an effective alternative to continuous therapy without cyclosporine. PMID- 15901985 TI - [Delayed graft function and its impact on the antigraft response after cadaver kidney transplantation]. AB - The purpose of this study was to evaluate the incidence of delayed graft function and its impact on the antigraft response after cadaver kidney transplantation. The analysis is based on 183 consecutive cadaver kidney transplantations performed in Vilnius University Hospital Santariskiu klinikos from January 2000 to December 2004. Delayed graft function occurred in 21.3% (39/183) of kidney transplantations. The frequency and severity of acute rejection episodes in recipients during first three months after transplantation and graft survival rate at one and two years were evaluated. Group 1 consisted of 39 patients with delayed graft function and group 2 (control group) of 144 patients with graft function immediately after transplantation. The maintenance immunosuppressive therapy consisted of cyclosporine, mycophenolate mofetil/azathioprine and prednisolone. The proportion of patients treated with monoclonal antibodies was similar in both groups (35.9% vs. 33.3%). Actuarial graft survival was estimated by the modified Kaplan-Meier method, graft loss was censored for death of recipient with functioning transplant and other causes of loss not related to rejection. There were no significant differences in the age of recipients (42.3+/ 11.3 vs. 39.4+/-14.1), as well as in HLA matching (2.2/6 M vs. 2.2/6 M), in the number retransplanted patients (10.3% vs. 10.4%) and in highly sensitized patients (plasma renin activity >50.0%) (5.1% vs. 4.8%) between those groups. Significant differences were observed in donors over 50 year (33.3% vs. 18.7%; p<0.05), in cold ischemic time over 20 h (53.8 vs. 32.6%, respectively). The occurrence of acute rejection episodes was higher in group 1 than in group 2 (69.2% (27/39) vs. 34.7% (50/144); chi2=14.9945, p<0.05). Graft survival was 88.5%, 84.3% at one year and two years in group 1 and 94.7%, 93.8% at one year and two years in group 2 (ns). Donor age >50, cerebral vascular disease as cause of donor death, and cold ischemic time >20 h are the main risk factors for delayed graft function. Delayed graft function is a risk factor for acute rejection episodes, but it has no impact on graft loss due to immunological reason at one and two years. These data may serve for tailoring immunosuppressive protocols. PMID- 15901986 TI - Survival of malignant pleural mesothelioma cases in the Tuscan Mesothelioma Register, 1988-2000: a population-based study. AB - This study analyses survival of Tuscan residents (Italy, 3.5 million population) diagnosed by histological examination with malignant pleural mesothelioma (MPM) during the period 1988-2000, and recorded in the Tuscan Malignant Mesothelioma Register. The aim was to establish the prognostic role of demographic, diagnostic and asbestos exposure variables. During 1988-2000, 381 MPM cases were recorded (318 men; 63 women). Vital status was ascertained up to 31 December 2002. No cases were lost to follow-up. Median survival of certain MPM was 324 days (11 months; 95% CI 297-366); 45.7% (95% CI 40.6-50.6%) survived more than 1 year; 24.2% (95% CI 20.0-28.5%) more than 2 years. In univariate and multivariate analyses survival was associated with histological subtype (epithelioid subtype had the longest survival). Gender, age, period of diagnosis, hospital of diagnosis and asbestos exposure did not show significant effects. Therapeutic information was available for patients of the period 1997-2000. There was no significant difference in survival between treated versus untreated patients. In conclusion, no advance in prognosis at the population level in the most recent period can be suggested on the basis of the data available to the Tuscan Malignant Mesothelioma Register. PMID- 15901987 TI - The data quality of haematological malignancy ICD-10 diagnoses in a population based hospital discharge registry. AB - The objectives of this study were to estimate the data quality of haematological malignancy diagnoses in a hospital discharge registry, and to quantify the impact of any misclassification of diagnoses on survival estimates. We included all patients > or = 15 years living in North Jutland County, Denmark with a first time discharge diagnosis of a haematological malignancy registered in the Hospital Discharge Registry and the Danish Cancer Registry, the reference standard, from 1994 to 1999. We estimated completeness and positive predictive value (PPV) of haematological malignancies and specific subcategories, as a measure of data quality, and compared mortality rates based on data from the two registries by Cox regression analysis. Completeness in the Hospital Discharge Registry for all haematological malignancies was 91.5% (95% confidence interval (CI) 89.6-93.1) and PPV was 84.5% (95% CI 82.2-86.5). Reviews of the pathological files showed misclassified cases in both registries and thus indicated that both completeness and PPV of the Hospital Discharge Registry were underestimates. Mortality rate ratio for all haematological malignancies when registered in the Hospital Discharge Registry compared with being registered in the Danish Cancer Registry was 0.98 (95% CI 0.88-1.09). Discharge data had some misclassifications but these had no major impact on survival estimates. PMID- 15901988 TI - Chemoprevention of aflatoxin B1-induced genotoxicity and hepatic oxidative damage in rats by kolaviron, a natural bioflavonoid of Garcinia kola seeds. AB - The chemopreventive effects of kolaviron, a natural antioxidant bioflavonoid from the seeds of Garcinia kola, on aflatoxin B1 (AFB1)-induced genotoxicity and hepatic oxidative damage was investigated in rats. Kolaviron administered orally at a dose of 200 mg/kg once a day for the first 2 weeks and then 100 mg/kg twice a day for the last 4 weeks of AFB1 (2 mg/kg, single dose, intraperitoneal) treatment reduced the AFB1-increased activities of aspartate amino transferase (AST), alanine amino transferase (ALT) and gamma glutamyltransferase (gamma-GT) by 62%, 56% and 72% respectively. Malondialdehyde (MDA) formation and lipid hydroperoxide (LHP) accumulation were observed in the livers of AFB1-treated rats. Kolaviron significantly reduced the AFB1-induced MDA and LHP formation. Vitamins C and E were protective in reducing the increase in the activities of AST, ALT and gamma-GT as well as lipid peroxidation caused by AFB1 (P<0.01). Administration of rats with kolaviron alone resulted in significant elevation in the activities of glutathione S-transferase, uridyl glucuronosyl transferase and NADH:quinone oxidoreductase by 2.45-, 1.62- and 1.38-folds respectively. In addition, kolaviron attenuated the AFB1-mediated decrease in the activities of these enzymes (P<0.01). Pretreatment of rats with kolaviron, vitamins C and E alone did not exert genotoxicity assessed by the formation of micronucleated polychromatic erythrocytes (MNPCEs) (P>0.05). Co-treatment of rats intraperitoneally with kolaviron (500 mg/kg) 30 min before and 30 min after AFB1 (1 mg/kg) administration inhibited the induction of MNPCEs by AFB1 (P<0.001) after 72 h. While vitamin C was effective in reducing AFB1-induced MNPCEs formation, vitamin E did not elicit any antigenotoxic response. These results indicate kolaviron as effective chemopreventive agent against AFB1-induced genotoxicity and hepatic oxidative stress. Thus kolaviron may qualify for clinical trial in combating the menace of aflatoxicosis in endemic areas of aflatoxin contamination of foods. PMID- 15901989 TI - Black tea polyphenols suppress cell proliferation and induce apoptosis during benzo(a)pyrene-induced lung carcinogenesis. AB - One of the most promising strategies for cancer prevention is chemoprevention by daily used food and beverages. Black tea, the most widely consumed beverage, is a source of compounds with antioxidative, antimicrobial, antimutagenic and anticarcinogenic properties. Lung cancer is the most common cause of cancer deaths in both men and women worldwide. Over one million people around the world are likely to be killed by lung cancer due to increased tobacco smoking and environmental pollutants, especially car exhausts. Therefore chemopreventive intervention using black tea and its active components may be a viable means to reduce lung cancer death. In the present investigation, we used benzo(a)pyrene (BP) to induce lung carcinogenesis in mice for the assessment of potential apoptosis-inducing and proliferation-suppressing effects of theaflavins and epigallocatechin gallate, active components of black tea. Hyperplasia, dysplasia and carcinoma in situ evident in the carcinogen control group on the 8th, 17th and 26th weeks respectively, were effectively reduced after treatment with theaflavins and epigallocatechin gallate. Significant reduction in number of proliferating cells and increased number of apoptotic cells was also found on the 8th, 17th and 26th week of treatment with theaflavins and epigallocatechin gallate in BP-exposed mice. Our observation suggests a promising role for black tea polyphenols in the prevention of lung cancer. PMID- 15901990 TI - Genetic analysis of microsomal epoxide hydrolase gene and its association with lung cancer risk. AB - The human microsomal epoxide hydrolase (EH) gene contains polymorphic alleles, which may be linked to increased risk for tobacco-related lung cancer. The purpose of this study is to screen new polymorphisms and determine whether these polymorphisms can be used to predict individual susceptibility to lung cancer. The polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis was used to screen for polymorphisms in the coding region of the EH gene. Eleven polymorphisms, including previously reported polymorphisms, were identified and the prevalence of these variants was assessed in at least 50 healthy Caucasians and African-Americans. Among the 11 polymorphisms, the prevalence of the amino acid-changing EH polymorphisms in codons 43, 113 and 139 was examined in 182 Caucasian incident cases with primary lung cancer, as well as in 365 frequency-matched controls to examine the role of EH polymorphisms in lung cancer risk. A significant increase in lung cancer risk was observed for predicted high EH activity genotypes (odds ratio (OR) 2.3, 95% confidence interval (CI) 1.2-4.3) as compared with low EH activity genotypes. This association was more pronounced among patients with lung adenocarcinoma (OR 4.7, 95% CI 1.7-13.1). These results suggest that the EH polymorphism plays an important role in lung cancer risk and is linked to tobacco smoke exposure. PMID- 15901991 TI - Reduction of clinically manifest colorectal cancer by endoscopic screening: empirical evaluation and comparison of screening at various ages. AB - Endoscopic screening (sigmoidoscopy, colonoscopy) with removal of precancerous lesions can prevent a large proportion of colorectal cancers (CRCs). However, there is lack of data regarding optimal age, time intervals and numbers of screening examinations. We developed and applied modified techniques of epidemiological analysis to evaluate the impact of various endoscopy-based screening strategies on prevention of clinically manifest CRCs between the ages of 50 and 79 in a population-based case-control study (294 cases, 254 controls) conducted in Saarland, Germany. We found a strong potential for reduction of CRC occurrence even with a single screening endoscopy. The optimal age for a single screening endoscopy appears to be around 55 (estimated potential for prevention of cases between the ages of 55 and 79 in case of 100% compliance: 77% (95% confidence interval (CI) 46-90%)). A single screening endoscopy at age 50 would have a lower impact due to failure to prevent CRC at higher ages. Similarly, screening at ages 60 or older would have a lower impact because it would fail to prevent CRC at lower ages. Repeated offers of screening examinations could provide substantial additional benefit with the levels of compliance to be expected in practice, but they would have to be weighed against the increased risks and costs. PMID- 15901992 TI - Intake of wine, beer and spirits and risk of gastric cancer. AB - The objective was to study prospectively the relation between quantity and type of alcohol and risk of gastric cancer. In a pooled database from three population studies conducted in 1964-1992, a total of 15,236 men and 13,227 women were followed for a total of 389,051 person-years. During follow-up 122 incident cases of gastric cancer were identified. Total alcohol intake itself was not associated with gastric cancer, but type of alcohol seemed to influence risk. Compared with non-wine drinkers, participants who drank 1-6 glasses of wine had a relative risk ratio of 0.76 (95% confidence interval (CI) 0.50-1.16), whereas those who drank >13 glasses of wine per week had a relative risk ratio of 0.16 (95% CI 0.02 1.18). Linear trend test showed a significant association with a relative risk ratio of 0.60 (95% CI 0.39-0.93) per glass of wine drunk per day. These relations persisted after adjustment for age, gender, educational level, body mass index, smoking habits, inhalation and physical activity. There was no association between beer or spirits drinking and gastric cancer. In conclusion, the present study suggests that a daily intake of wine may prevent development of gastric cancer. PMID- 15901994 TI - Results of breast cancer screening in first generation migrants in Northwest Netherlands. AB - To determine breast cancer screening results according to country of birth data were used from the breast cancer screening organization of the Comprehensive Cancer Centre Amsterdam, The Netherlands. Overall (age-adjusted) attendance of the breast cancer screening was 76% for women aged 50-69. Attendance was significantly lower for women born in non-western countries (Surinam 59%, Turkey 44% and Morocco 37%) and for women with residence in Amsterdam (68%). Referral and detection rates for women from non-western countries were 5.1 and 2.2 per 1000 screened women, respectively, compared with 8.8 and 4.0 for women born in The Netherlands (P<0.05). The positive predictive value was 45% for women born in The Netherlands and western countries and 43% for women born in non-western countries. Although women born in non-western countries attend breast cancer screening less frequently than women born in The Netherlands, they also have a low detection rate. The latter finding justifies a passive attitude towards the low attendance. PMID- 15901993 TI - Association between genetic polymorphisms of CYP1A2, arylamine N acetyltransferase 1 and 2 and susceptibility to cholangiocarcinoma. AB - Human CYP1A2 and arylamine N-acetyltransferases, which are encoded by the polymorphic CYP1A2 and NAT genes respectively, have been shown to have wide interindividual variations in metabolic capacity and may be potential modifiers of an individual's susceptibility to certain types of cancers. The present study aimed to evaluate the relationship between CYP1A2, NAT1 and NAT2 polymorphisms and cholangiocarcinoma (CCA), the most prevalent cancer in the north-east of Thailand. A total of 216 CCA patients and 233 control subjects were genotyped by polymerase chain reaction with restriction fragment length polymorphism based assays. Two CYP1A2 alleles (CYP1A2*1A wild-type and *1F), six NAT1 alleles (NAT1*4 wild-type, *3, *10, *11, *14A and *14B) and seven NAT2 alleles (NAT2*4 wild-type, *5, *6A, *6B, *7A, *7B and *13), which are the major alleles found in most populations, were analysed. Although CYP1A2*1A allele, NAT1*10 allele, and the NAT2 slow acetylator alleles were not associated with CCA risk, among the male subjects, the genotype CYP1A2*1A/*1A conferred a decreased risk of the cancer (adjusted odds ratio (OR) 0.28, 95% confidence interval (CI) 0.08-0.94) compared with CYP1A2*1F/1*F. Frequency distributions of rapid NAT2*13 and two slow alleles (*6B and *7A), but not the other major alleles, were associated with lower CCA risk. Adjusted OR of the genotypes consisting of at least one of these alleles significantly decreased the cancer risk compared with none of them (OR 0.26, 95% CI 0.15-0.44). This study suggests that the NAT2 polymorphism may be a modifier of individual risk to CCA. PMID- 15901995 TI - Genetic variants of myeloperoxidase and catechol-O-methyltransferase and breast cancer risk. AB - This nested case-control study evaluated the role of polymorphisms in the myeloperoxidase (MPO) and catechol-O-methyltransferase (COMT) genes that modulate oxidative stress in breast cancer risk in a Chinese population. Our results demonstrate that the MPO A/A genotype was associated with a reduced risk of breast cancer (odds ratio (OR) 0.64; 95% confidence interval (CI) 0.11-3.76), whereas there was no overall association of COMT genotype with breast cancer. Of note, an elevated breast cancer risk associated with the increasing numbers of high-risk genotypes of MPO and COMT genes was observed in women with a longer duration between menarche and first full-term pregnancy. PMID- 15901996 TI - Exogenous supplementation with omega-3 polyunsaturated fatty acid docosahexaenoic acid (DHA; 22:6n-3) synergistically enhances taxane cytotoxicity and downregulates Her-2/neu (c-erbB-2) oncogene expression in human breast cancer cells. AB - Omega-3 polyunsaturated fatty acid (PUFA), docosahexaenoic acid (DHA; 22:6n-3) and other omega-3 and omega-6 PUFAs have raised interest as novel anticancer agents by exerting selective cytotoxic effects on human cancer cells without affecting normal tissues. Here, we examined the in vitro relationship between exogenous supplementation with DHA and breast cancer chemosensitivity to taxanes. We measured cell viability in the highly metastatic human breast cancer cell line MDA-MB-231 exposed sequentially to DHA followed by paclitaxel (Taxol) or docetaxel (Taxotere). As DHA by itself showed cytotoxic effects, possible synergistic interactions between DHA and taxanes were assessed, employing the combination index (CI) method and the isobologram analysis. Both methods showed a strong synergism (CI approximately 0.5; P<0.005) between DHA and taxanes in MDA MB-231 cells. When the increase in taxanes efficacy was measured by dividing the IC50 values (50% inhibitory concentrations) obtained when the cells were exposed to taxanes alone by those after DHA pre-exposure, we found that DHA enhanced the cytotoxic activity of taxanes against MDA-MB-231 cells in a dose-dependent manner (up to 13- and 5-fold increase in Taxol and Taxotere efficacy, respectively). Importantly, sequential exposure to DHA followed by taxanes also yielded strong synergism in Her-2/neu (c-erbB-2)-overexpressing and taxanes-resistant SK-Br3 and BT-474 breast cancer cells. Moreover, exogenous supplementation with DHA significantly decreased the expression of Her-2/neu-codified p185(Her-2/neu) oncoprotein (up to 78% reduction in BT-474 cells). Our results provide experimental support to the hypothesis that omega-3 PUFAs can be used as modulators of tumor cell chemosensitivity and provide the rationale for in vivo preclinical investigation. In addition, this is the first study demonstrating that omega-3 PUFA DHA downregulates Her-2/neu oncogene expression in human breast cancer cells. PMID- 15901997 TI - Self-reported diagnostic X-ray investigation and data from medical records in case-control studies on thyroid cancer: evidence of recall bias? AB - The aim of the present investigation was to evaluate and compare data from medical records with mailed questionnaires concerning risk factors for diagnostic X-ray investigation induced thyroid cancer. This study encompassed 180 patients and 360 controls, selected from the National Population Registry. Information on medical X-ray exposure was obtained by medical records and also from mailed questionnaires, with a latency of 5 years from diagnosis. The mailed questionnaire was answered by 171 cases (95%) and 325 controls (90%). In 132 cases and 251 controls data on medical records were collected and compared with data from mailed questionnaires for the same subjects. No earlier X-ray investigation based upon medical records was reported in 39 cases and in 74 controls. The median cumulative thyroid dose for cases was 1.1 mGy in medical records and in mailed questionnaire 0.6 mGy. For the controls the respective median cumulative dose was 1.0 mGy in medical records and 0.3 mGy in mailed questionnaires. The median ratio between medical records and mailed questionnaires in the cases was 2.1. For the controls the median ratio was 2.9. Wilcoxon's Matched Pairs Test (WMPT) showed a significantly underreport of thyroid X-ray dose in both cases (P<0.01) and controls (P<0.000001). In cases younger than 50 years at the time of the diagnosis of thyroid cancer there was not a significant underreport of X-ray examinations. Corresponding data from the control group showed however a significant underreport. Both cases and controls older than 50 reported significantly fewer X-ray investigations compared with data from medical records. A significant underreport was found among women both in cases and controls. For men there was a slight underreport among both cases and controls, although not significant. In conclusion, when studying diagnostic X ray investigation as a risk factor for thyroid cancer, it is important to reduce the potential for recall bias when the study relies only on case-control reporting. To complement a case-control study with prospective medical data recorded at the time of the investigation could be an appropriate way to reduce the risk for recall bias. PMID- 15901998 TI - GSTO polymorphism analysis in thyroid nodules suggest that GSTO1 variants do not influence the risk for malignancy. AB - A new class of glutathione S-transferase enzymes named omega (GSTO) has been recently identified and shown to be expressed in a wide range of human tissues. A genetic polymorphism of the GSTO1 gene causing an alanine-to-aspartate (A140D) substitution in amino acid 140 produces a variant with lowered enzyme activities in the biotransformation of inorganic arsenic, a common contaminant of drinking water in many regions of the world and a well-known carcinogen. In order to investigate the role of GSTO1 inheritance pattern on thyroid cancer risk we used a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) sequencing approach to compare the genotypes of 173 (87 women, 86 men; 18-81 years old; 47+/-18 years old) healthy control individuals with those of 145 patients with thyroid nodules (84 women, 61 men; 17-81 years old; 49+/-14 years old) including 17 follicular carcinomas, 76 papillary carcinomas, 21 follicular adenomas and 31 multinodular goiters. The incidence of GSTO1 variants was similar in the control population and population with the benign and malignant nodules. There was no association between genotype and the patients' clinical features, tumour parameters of aggressiveness at diagnosis or behaviour during follow-up. We conclude that GSTO1 variants do not influence the risk for thyroid nodules or their pathologic and clinical characteristics. PMID- 15901999 TI - Glutathione S-transferase M1 and T1 null genotype frequency in chronic myeloid leukaemia. AB - Polymorphisms associated with genes coding for glutathione S-transferase enzymes are known to influence metabolism of different carcinogens and have been associated with incidence of various types of cancer. We have determined the GST M1 and GST T1 'null' genotype frequency in 81 patients with chronic myeloid leukaemia (CML) and 123 racially and geographically matched control individuals by multiplex polymerase chain reaction (PCR). GST M1 null genotype frequencies in CML and controls were 28.4% and 27.7%, respectively. GST T1 null genotype frequencies in CML and controls were 19.8% and 7.3%, respectively. The GST T1 null genotype frequency in CML patients is significantly different from that in controls (odds ratio (OR) 3.12, 95% confidence interval (CI) 1.3-7.45, P=0.008). PMID- 15902000 TI - How to combine the use of different mobile and cordless telephones in epidemiological studies on brain tumours? AB - Mobile phone users in epidemiological studies have often used more than one phone model, and sometimes also more than one mobile phone system (analogue and digital systems). Until now, this has not been taken into account in epidemiological studies, mainly because we do not know the possible interaction mechanism(s) and, hence, how to integrate exposure from different phones into one dosimetric measure. In this paper we take a step towards starting a discussion about how to proceed with this important issue and the possible use of parameters such as weighting factors, measured specific absorption rate (SAR) values and integrated specific absorption values are discussed. As a base of this discussion two previously published studies are used, one on mobile phones and cancer and the other one on subjective symptoms. PMID- 15902001 TI - No considerable long-term weight gain after smoking cessation: evidence from a prospective study. AB - The aim of the study was to analyse predictors of the body mass index (BMI) after smoking cessation. The sample included 4075 residents aged 18-64 years in a northern German area (participation rate 70.2%), drawn randomly from the resident registration files, among them 1545 current and 903 former daily smokers. The current smokers were followed up 36 months after baseline. Face-to-face in-home computer-aided interviews (Composite International Diagnostic Interview) including questions about body weight and height at baseline and postal questionnaires at follow-up were employed. The data reveal that the number of cigarettes smoked at time of peak consumption in life contributed substantially, and years of abstinence from daily smoking contributed marginally to the BMI in a general linear model. It is concluded that the contribution of smoking cessation to the BMI increase was practically negligible. Efforts to prevent weight gain after smoking cessation should focus especially on heavy smoking. PMID- 15902002 TI - Cancer mortality during the 1968-1994 period in a mining area in France. AB - We performed a geographical analysis of cancer mortality in the communes surrounding an industrial mining complex (Salsigne, France) where suspicious levels of pollution due to arsenic were measured. Compared with that observed in a control area, we showed a significant excess of mortality due to all cancer types (ratio of standard mortality ratios (ratio of SMRs)=1.1), lung cancer (ratio of SMRs=1.8), pharynx cancer (ratio of SMRs=2.1) in the whole population, and due to digestive system cancer (ratio of SMRs=1.3) among women. The results were similar after controlling for the occupation distribution in the populations. Excluding mining complex workers deaths from the deaths in the studied populations did not modify the pattern of our results. We concluded that the excess of cancer deaths could not be exclusively due to potential professional exposures among the workers of the mining complex and are probably explained by environmental contamination. PMID- 15902003 TI - Glass punching. PMID- 15902006 TI - Skeletal pin site care: National Association of Orthopaedic Nurses guidelines for orthopaedic nursing. AB - A systematic analysis of the research literature on skeletal pin site care was conducted, and the opinions of an expert panel were obtained. Four specific recommendations for skeletal pin site care are offered, with explicit discussions of the level of research support and/or expert panel support for each. Discussion of other pin site care issues is provided, and characteristics of the research base regarding skeletal pin site care are described. PMID- 15902007 TI - Help! My orthopaedic patient is pregnant! AB - Caring for a pregnant patient is a possibility in every field of nursing. Understanding the effects various illnesses may have on pregnancy and vice versa is important in preventing damage to the fetus and/or complications for the pregnant woman. This article assists orthopaedic nurses to practice proactively and anticipate problems during the care of the pregnant patient and avoid undesirable outcomes. Selected physiologic changes in pregnancy and their implications of orthopaedic conditions are described. Implications for nursing care with pregnant women with an orthopaedic condition are also discussed. PMID- 15902009 TI - Attitudes toward advance directives and advance directive completion rates. AB - BACKGROUND: The Patient Self-Determination Act (PSDA) (part of the ) requires that all healthcare institutions receiving Medicare and Medicaid funds inform patients about their right to participate in healthcare decisions, including their right to have an advance directive. Advance directives (ADs) allow an individual to participate indirectly in future medical care decisions if he or she becomes decisionally incapacitated. Despite passage of this bill and mechanisms within most healthcare institutions to provide this information, the AD completion rate remains low. PURPOSE: The purpose of this study was to identify the significant attitudes and factors that influence the completion of ADs among adult hospitalized patients admitted to medical/surgical units. DESIGN: Descriptive correlational survey design. METHODS: A convenience sample of patients admitted to medical-surgical units during a 2-month period completed face-to-face interviews and completion of the Advance Directive Attitude Survey. FINDINGS: Eighty-two percent of the sample identified having received information on ADs, although hospital policy had it distributed to all patients on admission. Eighteen percent of the sample had completed an AD, and an additional 8% completed an AD after the interview and accompanying education during the interview for a 26% completion rate. The mean attitude toward ADs was slightly positive, and there was no difference in attitude score between those with an AD and those without. IMPLICATIONS: Low completion rates of ADs among the majority of the population and even lower among ethnically diverse individuals despite favorable attitudes toward ADs suggest that there are factors beyond access to information that may influence the decision not to complete an AD. Results of this study are congruent with other research raising the question of whether ADs as currently designed are appropriate for all groups. PMID- 15902011 TI - One vision, one voice: transforming caregiving in nursing. AB - A clear vision is a powerful tool that will transform the future of nursing. Consumers have become more knowledgeable about their healthcare needs and more demanding of quality care and accountability from their healthcare providers. This consumer savvy is providing nursing with an incredible opportunity that must not be lost. This vision statement has the potential to empower the nursing profession and make that difference. PMID- 15902012 TI - Use of the Internet for health information by older adults with arthritis. AB - BACKGROUND: Despite the information available on the Internet and the increasing number of older adults who use the Internet, little is known about Internet use for health information by elders with arthritis. PURPOSE: The purposes of this study were (1) to examine the use of computers and the Internet by elders with arthritis and (2) to describe demographic and illness-related characteristics of elders who use the Internet to find health information. SAMPLE: Seventy-one elders with arthritis completed survey questionnaires. FINDINGS: Twenty-eight percent had a computer in their home, of whom 95% had Internet access. Thirty nine percent sought arthritis information on the Internet. People with more education were more likely to use the Internet. However, age and functional disability resulting from arthritis were not associated with Internet use. IMPLICATIONS: Health professionals need to be aware of Internet resources and assist elders in finding appropriate Internet-based arthritis information. PMID- 15902013 TI - HIPAA--Implications for research. AB - Privacy, anonymity, and informed consent are the hallmarks of current research conduct. How do the Health Insurance Portability and Accountability Act regulations regarding individually identified health information and protected health information affect research? The purpose of this article is to discuss ways that the Health Insurance Portability and Accountability Act is influencing the conduct of research, including the implications for institutional review boards, recruitment of subjects, obtaining consent, access to data, de identification of data, authorization to disclose data, and the processing, transmission, and storage of collected data. PMID- 15902014 TI - Wound healing. AB - Wound healing in orthopaedic care is affected by the causes of the wound, as well as concomitant therapies used to repair musculoskeletal structures. Promoting the health of the host and creating an environment to foster natural healing processes is essential for helping to restore skin integrity. Normal wound healing physiologic processes, factors affecting wound healing, wound classification systems, unique characteristics of orthopaedic wounds, wound contamination and drainage characteristics, and potential complications are important to understand in anticipation of patient needs. Accurate wound assessment and knowledge of nursing implications with specific wound care measures (cleansing, debridement, and dressings) is important for quality care. New technologies are enhancing traditional wound care measures with goals of effective comfortable wound care to promote restoration of skin integrity. PMID- 15902016 TI - Dupuytren's disease. AB - Dupuytren's disease is a nonmalignant fibroproliferative disease that causes progressive and permanent contracture of the palmar fascia with subsequent flexion contracture of the digits. Although the exact etiopathology in the development of Dupuytren's disease is unknown, certain familial, racial, and physiologic factors have been determined. The latest prospective pathogenesis of Dupuytren's disease are presented. PMID- 15902019 TI - Signs and symptoms of hypothetical wound assessment by nurses. AB - This is a report of a study in which tissue viability nurses, MSc student nurse practitioners, and post-registration nurses on a wound management module (n = 16) ranked signs and symptoms of wound healing, stasis and deterioration relative to their supposed importance. The top ranked sign for a healing wound was 'size' (reduction), for a static wound it was 'no' (change), and for a deteriorating wound 'increase' (pain). This was a convenience sample, and caution is indicated in generalizing results. However, there were statistically significant differences between assessors in the number of words they used (ANOVA P < 0.001), and in the words they used for different wound phases (chi(2)) P < 0.001). The study supports the view that there are some words used in common by different respondents in specific wound phases. However, a previous study raised questions about what people actually mean when they use a particular word (Maylor, 2003). PMID- 15902020 TI - Pressure ulcer benchmarking within a primary care setting. AB - Patient-focused benchmarking was initially launched by the Department of Health. This article examines how the tissue viability service of Camden and Islington primary care trusts implemented the pressure of ulcer benchmarking process within the nursing teams of these trusts. This was achieved by: agreeing best practice through examination of local and national guidelines; developing a suitable audit tool for nursing teams to assess their clinical areas against this best practice; nursing teams producing core action plans to facilitate movement towards best practice; disseminating results; reauditing. The results of the audits carried out in October 2003 and February 2004 are presented. A plan for future work is described including: the involvement of other members of the multidisciplinary team and the Patient Advice Liaison Service; linking pressure ulcer benchmarking to the benchmarks of communication, privacy and dignity and record keeping. PMID- 15902021 TI - Election fever: TVNs must make their voice heard. PMID- 15902022 TI - Changing pressure-redistributing mattress stocks: costs and outcomes. AB - There remains considerable confusion regarding the selection of appropriate pressure-redistributing support surfaces, although it is accepted that use of low pressure foam mattresses is likely to reduce the incidence of pressure ulcers compared with standard hospital mattresses. In this study, a 650- hospital replaced its mattress stock with low-pressure foam mattresses with over pounds sterling 100 000 cost savings in the first year after implementation. Incidence and prevalence data were recorded but given the challenges of interpreting apparent trends in the data (due to the lack of robust data collection methodologies and no case-mix adjustment) these data were not included. PMID- 15902023 TI - Clinical variance in assessing risk of pressure ulcer development. AB - Nurses working in one area health service (AHS) in Sydney, Australia, expressed concern about the development of pressure ulcers in hospitalized patients. Anecdotal evidence suggested that a variety of approaches were being used to assess patients to identify those at risk of pressure ulcer development. A questionnaire was distributed to all registered nurses (n = 2113) in clinical settings within the AHS. Data were analysed using frequency distribution. The response rate was 40% (n = 850), of which 444 were useable. Nurses generally do not use a tool to assess pressure ulcer risk potential, but rely on a range of practice procedures and risk indicators. It is recommended that a pressure ulcer project group be established to evaluate existing tools or, if necessary, develop a tool for the assessment of patients to identify those at risk of developing pressure ulcers. PMID- 15902024 TI - BMA claims that patients in A&E are being put at risk. PMID- 15902025 TI - Palliative care: challenges in caring for dying people. PMID- 15902026 TI - The pre- and postoperative nursing care for patients with a stoma. AB - This article revisits the various issues that surround the nurse caring for a patient with a stoma (colostomy, ileostomy or urostomy). Understanding the basic anatomy, reasons for stoma formation and the surgical operations, including variations such as continent stomas, can assist the nurse in caring for this patient group. The article discusses briefly the various stoma appliances that can be used by the nurse or ostomate (person with a stoma). Practical tips on pre and postoperative nursing care of the patient with a newly formed stoma are provided for the nurse. In some cases, unfortunately, complications may occur following stoma-forming surgery; these are discussed and nursing advice provided. One of the most important ways in which the nurse can support the patient is to teach the patient his/her stoma care, ensuring independence before discharge and showing empathy and compassion. PMID- 15902027 TI - Teaching stoma-management skills: the importance of self-care. AB - Patients undergoing stoma formation have to make major physical and psychological adjustments following surgery. Research indicates that the early promotion of stoma-management skills is, therefore, a critical concern for the hospital-based stoma care nurse. This article aims to explore how the stoma care nurse specialist can best utilize teaching opportunities in the pre- and post-operative periods. Teaching should be approached in an organized manner, underpinned by sound principles of teaching and learning using teaching plans where appropriate to ensure that no vital aspects are omitted. Realistic learning outcomes should be established in consultation with the patient according to his/her readiness and ability to learn. An organized exchange of information between hospital and community will ensure that teaching and support are continued following discharge from hospital promoting positive adjustment to stoma formation and thus facilitating successful rehabilitation. PMID- 15902028 TI - A new flangeless adhesive coupling system for colostomy and ileostomy. AB - A large clinical study evaluated the performance and patient acceptance of a flangeless stoma system - the Esteem synergy system (ConvaTec). Key parameters for previous one-piece users were product versatility assessed by ease of application/removal, security, convenience of use and overall subject preference. Key parameters for previous two-piece users were product flexibility assessed by profile, comfort, rigidity, security, and overall subject preference. Wear time and safety were also assessed. In 44.5% of one-piece users the test product was rated at least as good as their usual product. In two-piece users the test product was statistically significantly better than their usual product in terms of flexibility and was rated at least as good as their usual system by 56.3% of subjects. The overall mean wear time was approximately 2 days. The test system was well tolerated. In summary, Esteem synergy successfully combined the attributes of both one- and two-piece stoma systems. PMID- 15902029 TI - Benefits of narrative therapy: holistic interventions at the end of life. AB - Clinical experience tells us that the activity of 'putting the house in order' as patients reach the terminal stage in their illness is both a common and legitimate exercise. This process can entail many activities, both physical and emotional. For example, arranging financial affairs, making a will, compiling a living will, organizing their own funeral, completing unfinished emotional business, and perhaps most importantly, pondering the very meaning of life itself. In a continual search to find innovative ways of supporting those who are touched by death and dying, this article will discuss how narrative therapeutic approaches could be helpful in supporting the family of a terminally ill patient who is 'putting the house in order'. Incorporated within this specialist area are perceptions of death and dying, theories of death and dying as well as emotions and the search for meaning. Also under review will be the evidence base for psychotherapeutic interventions, and a review of the narrative therapy approach. PMID- 15902030 TI - Exploring the financial provision available following death. PMID- 15902031 TI - Staff nurse who was drug-dependent and stole from the ward stock. PMID- 15902032 TI - Nurse leadership, interprofessionalism and the modernization agenda. AB - As part of the drive to deliver effective health and social care, the Government's modernization agenda focuses on strengthening nursing leadership and developing interprofessional collaboration. Senior nurses are well placed to promote the latter by supporting substantive change in NHS culture. Findings from a research study, conducted by the Faculty of Health and Social Care, University of the West of England, Bristol, as part of a research programme evaluating its interprofessional curriculum, have highlighted senior nurses' potential contribution to implementing and maintaining effective interprofessional collaboration. The authors argue that senior nurses are particularly well placed to help students across disciplines gain the experience needed to develop the requisite skills for delivering care within interprofessional services; and that the implementation of roles and initiatives which support senior nurses in developing appropriate leadership skills is a key component underpinning the modernization agenda. PMID- 15902033 TI - Older people in acute hospitals need more care. PMID- 15902034 TI - Research shows how we can prevent falls in old age. PMID- 15902035 TI - Review of mental health nursing is long overdue. PMID- 15902036 TI - A nurse-led ear care clinic: sharing knowledge and improving patient care. AB - This article covers the nursing role within an ear care clinic in the hospital setting. The range of conditions managed in the clinic are listed and some of the current issues affecting ear care and ear, nose and throat (ENT) are discussed. The author conveys her enthusiasm for the appropriate management of wax encouraging colleagues to appreciate the negative affect wax impaction can have on patients. Hearing impairment is briefly mentioned and a screening list is provided to assess the hearing of older adults. Patient care is discussed and there are suggestions for the improvement of the provision of ear care within the community. The nurse practitioner discusses how to facilitate the sharing of knowledge with community practitioners. Study seminars, publications, the use of the hospital intranet site, treatment and discharge summaries can all be used to communicate new developments in patient care. PMID- 15902037 TI - The nurse practitioner in primary care: alleviating problems of access? AB - Improving access to primary care services is an essential component of the NHS modernization plan and the advent of independent nurse practitioners in primary care has focused attention on the extent to which this group of nurses can effectively substitute for GPs. This study was designed to explore the role of a nurse practitioner in primary care, particularly whether the provision of a nurse practitioner facilitated access to care that met the needs of patients. Semistructured interviews were conducted with 14 patients who had consulted with the nurse practitioner, 10 staff within the practice who had knowledge of the role, and the nurse practitioner herself. With the permission of interviewees, interviews were audiotaped, the tapes transcribed verbatim, and the data were coded by theme. It was perceived by both groups of interviewees that access to care had been improved in that there were more appointments available, appointments were longer than they had been previously and were available at different times of the day. However, some areas in which access was 'restricted' were articulated by staff interviewees, such as limitations to the nurse practitioner's prescribing and problems with referring patients to secondary care. Additionally, while access to a member of the primary healthcare team was improved for many patients, access to a specific member of the team, such as a GP, was not always improved. Concerns were also expressed about how the role of the nurse practitioner needed to be developed in the practice. It can be concluded from this study that, potentially, the role of nurse practitioner has much to offer in terms of addressing problems of access in primary care for some patients. However, this is not a straightforward solution and in order for the role to be effective several issues highlighted in this study require addressing. PMID- 15902038 TI - A nurse-led clinic in chronic and allergic contact dermatitis. AB - It has generally been acknowledged that the incidence and prevalence of contact dermatitis has increased in recent years. It can develop at any stage of a person's life span (rare before puberty), and in many different circumstances and occupations. The demand on consultant dermatologists for contact dermatitis services has resulted in the depletion of some essential components to the investigation process, and patient outcomes have been affected. In recent years it has also resulted in the development of nurse-led contact dermatitis services. The traditional role of the dermatology nurse in contact dermatitis is to carry out patch testing. Nurse-led services require the wider acquisition of skills and knowledge in relation to contact dermatitis. This article is an attempt to introduce the complexities of contact dermatitis, and nurse-led services. PMID- 15902039 TI - Matron who continued to make numerous drug administration errors. PMID- 15902040 TI - Importance of the appropriate selection and use of continence pads. AB - Urinary incontinence is becoming an increasingly common problem for older men and women living in the community. It can have a deleterious effect on quality of life and, although advances have been made in treatments and therapies for this condition, there is still confusion over selection of continence products. This article will explore the problems associated with product selection and discuss alternative advice. PMID- 15902041 TI - Blood transfusion and Jehovah's Witnesses: the legal and ethical issues. AB - A critical incident is any event that had an effect, or could have had an effect, on the welfare of a patient. Patients must be treated with respect by staff who demonstrate that they are sensitive to individual needs, values, beliefs and cultural background. This article will examine the legal and ethical issues relevant to the right of an unconscious Jehovah's Witness in an intensive care unit (ICU) to refuse a blood transfusion. The concepts of consent, capacity to consent, necessity to act, advance directives, decision making, consequences of failing to obtain consent, ethical principles, human rights and the best interests of the patient will be explored, in the light of relevant statute and case law, in order to demonstrate some of the legal and ethical complexities within acute healthcare delivery. PMID- 15902042 TI - The legal action that can be taken following a person's death. PMID- 15902043 TI - The principles of inflammation in the development of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory condition that involves all elements of the immune response. The aetiology of RA is complex and centres on the development of autoantibodies and immune complexes. The pathogenesis is multistage and involves cytokines, angiogenesis and rheumatoid factor. Nurses managing patients who suffer with RA need to be aware of the pathological changes involved in the disease and its contribution to the progression of the condition. The diagnosis of RA involves blood screening for rheumatoid factor, raised erythrocyte sedimentation rate (ESR), use of arthroscopy to provide evidence of histological changes in the synovium, presenting symptoms, and changes on X-ray. Patient assessment will consider both patient-specific and disease-specific variables, including evidence of non-articular manifestations of RA. Long-term care revolves around trying to maintain patient mobility and the protection of unaffected joints, monitoring for the side-effects from medication and progression of the disease or development of non-articular manifestations. PMID- 15902044 TI - Nurses' attitudes to evidence-based practice: impact of a national policy. AB - Clinical effectiveness and evidence-based practice should be key cornerstones of current nursing practice. This study used postal questionnaires to explore the impact of a national initiative on nurses' attitudes, knowledge and practice relating to these twin concepts. Results indicated that attitudes can be influenced by national policy initiatives. Influencing behaviour, however, appears to be more difficult; no change was seen in reported application of evidence-based practice. Nurses did, however, report a desire to increase their implementation of evidence-based practice; reasons for not doing so included lack of the necessary skills and time in the working day. It is recommended that these education and training needs must be addressed by allocating sufficient protected learning time and funds. A more difficult issue relates to the burden of clinical work, which may also be preventing greater uptake of evidence-based practice. PMID- 15902045 TI - The benefits of Tubifast Garments in the management of atopic eczema. AB - Atopic eczema is a chronic inflammatory condition affecting mainly children and is one of the most common skin disorders in the Western world. It has a strong genetic component associated with atopic conditions such as asthma, hayfever or urticaria. The onset is usually within the first year of life and it affects mainly the face, after which the pattern often changes to involvement of the flexures. The most prominent characteristic of atopic eczema is pruritus (itch) together with dry skin. An important treatment of atopic eczema is the application of emollients (moisturizers) to rehydrate and restore the barrier function of the skin. Wet wrapping using large quantities of emollients is a well recognized therapy for atopic eczema and other dry skin conditions. The introduction of new Tubifast Garments by Medlock Medical makes this topical therapy easy to apply for parents and carers in the management of children's eczema. PMID- 15902046 TI - More thought should be given to the team approach. PMID- 15902047 TI - [Outcome of isolated small bowel transplantation in adults: experience from a single Italian center]. AB - AIM: Isolated small bowel transplantation is becoming the treatment of choice for adult patients with serious parenteral nutrition (PN) related complications: we report our three-year experience (December 2000-December 2003) from a single Italian center (Modena-Italy), with one of the larger European series. METHODS: We transplanted 14 patients, with a previous mean PN course of 27 months and a mean 21-month post-transplantation follow-up (range 3-36 months), obtaining a one year actuarial survival rate of 92.3% with no intraoperative deaths. RESULTS: We lost 1 patient (7.2%), died for post-transplantation overwhelming sepsis following Cytomegalovirus (CMV) enteritis. Thirteen patients are alive, with one year actuarial graft survival rate of 85.1%: 1 patient underwent graft removal (7.2%) for intractable severe acute rejection. Our immunosuppressive regimen was based on tacrolimus and 3 induction protocols: daclizumab (8 patients) with steroids, alemtuzumab (4 patients) and thymoglobulin (2 patients) without steroids. In 9 cases, we added sirolimus. Nine recipients experienced 22 episodes of acute cellular rejection (ACR), treated successfully in all cases but one. One patient (7.2%) was treated successfully for Post Transplant Lymphoproliferative Disease (PTLD) and is disease-free after 8 months. CONCLUSIONS: Small bowel transplantation can achieve optimal results depending on appropriate immunosuppressive management and candidate selection, added to shorter ischemia time and careful donor and graft selection. PMID- 15902048 TI - [Long-term results of neoadjuvant radiochemotherapy in squamous carcinoma of the thoracic esophagus]. AB - AIM: Surgery is considered the mainstay of therapy for clinically resectable esophageal cancer, even though neoadjuvant treatments are frequently added. The aim of this study was to analyse our experience on neoadjuvant treatment of squamous cell carcinoma of the thoracic esophagus with special reference to long term RESULTS: METHODS: The results of 66 patients who underwent neoadjuvant chemo-radiotherapy for squamous cell carcinoma of the thoracic esophagus at the 1(st) Division of General Surgery, University of Verona, from February 1995 to December 2002 were analysed statistically. The median follow-up period for the surviving patients was 65.3 months. RESULTS: The induction treatment was completed in 93.9% of cases, with a null treatment related mortality and a complication rate of 34.8%. Sixty-one out of the 66 patients (92.4%) underwent resection with a R0-resection rate of 83.9%. A major pathological response (responders) was gained in 42.6% of the cases, with a complete response (pTONO) observed in 29.5% of the cases. Overall 5-year survival for the 66 patients was 30%, while the 5-year survival rate raised to 43% in R0-patients. A better long term survival was observed for responders with respect to ''non-responders'' with a 5-year survival rate of 70% and 13%, respectively (P<0.001). CONCLUSIONS: This neoadjuvant protocol regimen represents a feasible treatment with an acceptable morbidity. The tumor efficacy in term of pathological responses was similar to literature RESULTS: An high rate of R0-resections was achieved with a possibility of cure limited to this group of patients. A better long-term survival was observed in patients with major pathological responses. PMID- 15902049 TI - [The choice of surgical therapy in adenocarcinoma of the cardia]. AB - AIM: From 1996 the adenocarcinoma of the esophago-gastric junction (AEG) is divided into 3 types according to Siewert's classification. For AEG type I and III the surgical treatment is codified, while for type II is still controversial. The aim of our study is to understand what is the better surgical treatment for AEG type II. METHODS: From 1990 to 2002 we have performed 111 resections for adenocarcinoma of the cardia: 25 for AEG type I (all esophago-gastric resection), 39 for type II (22 esophago-gastric resection, 17 extended total gastrectomy with esophageal resection) and 47 for type III (8 esophago-gastric resection, 39 extended total gastrectomy with esophageal resection). RESULTS: The morbidity and mortality rates are 17 and 5.4%, without significant difference between the different surgical treatment (p>0.01). The 5 year survival rate is 35%. Significant prognostic factors are the staging TNM (p=0.002) and principally the presence of metastatic lymph nodes (p=0.001). For AEG type II any significant difference in survival is associated with surgical strategy, also in early stage (p>0.01). CONCLUSIONS: According to the results of our study and those of the other authors, who have showed that a 10 cm distance of the neoplasm by the gastric side and the esophageal one could assure oncologic radicality and also that metastatic lymph nodes below pylorus and near greater curvature are uncommon, we can consider esophago-gastric resection for AEG II a speedy, safe and oncologically correct surgical treatment. PMID- 15902050 TI - [Criteria for training in laparoscopic gastric surgery: guidelines and experience of 30 cases]. AB - AIM: Although many studies on laparoscopic surgery of the stomach have been conducted so far, yet they have not provided surgeons with criteria for gradual and safe training with this technique. The results of gastric surgery with 30 patients operated on by laparoscopic approach are hereby described. The aim of this issue is to provide surgeons with guide lines for progressive training, respectful to patients, complying with oncologic criteria and useful to reduce conversion rate or drawbacks at the start of the experience. METHODS: The Authors made a retrospective analysis on 30 patients affected by gastric lesions, 5 benign chronic ulcers and 25 neoplasms of the stomach. Our guide lines suggest that the training begin with the treatment of benign lesions, followed by early gastric cancer (EGC) and by advanced gastric cancer (AGC) of the antrum. Our experience started with 4 laparoscopic subtotal distal gastrectomies (LSGs) for benign ulcer; independent of the guidelines hereby proposed 1 laparoscopic total gastrectomy (LTG) was done after the intraoperative finding of a benign ulcer of the lesser curve penetrating into the left hepatic lobe. The beginning of training included also 1 LSG for distal stromal tumor (GIST). Subsequently 13 early gastric cancers (EGC) were operated on: echoendoscopy could demonstrate 12 T1 m and 1 T1 sm and no evidence of nodal involvement. The diameter of EGCs was 1,3 cm on average ( range 0,7-4 cm), all were marked by Indian ink to allow performance of 10 LSGs and 3 LTGs. Moreover, 8 LSGs for advanced gastric carcinoma (AGC) of the antrum were carried out. The training in malignancies progressed with LTG for 2 non-Hodgkin gastric lymphomas; 1 lymphoma required conversion to laparotomy due to infiltration of the diaphragmatic crus. A D2 lymphadenectomy was associated to gastrectomy in adenocarcinomas. RESULTS: The feasibility of laparoscopic gastric surgery was confirmed by this study, with operating time of 240 minutes (range 150-360), intraoperative blood loss was 180 ml (range 100-250), and only 1 patient required blood transfusion for postoperative bleeding. The specific morbidity rate was 10% owing to duodenal leakage in 3 cases in the early phase of this study (3/30): 1 required laparotomy. The mortality rate was 3% due to 1 serious postoperative bleeding and acute hepatic failure in a patient with post-alcoholic cirrhosis. The conversion rate was 3% (1/30). The nasogastric tube was removed on the 4(th) postoperative day, and the oral intake started on the 6(th) postoperative day after a barium follow-through examination. The mean postoperative hospital stay was 16 days (range 10-25). The number of nodes retrieved was 18 on average and it improved with the experience: from the minimum of 9 nodes in benign ulcers, it grew to 20 in EGCs and to 25 in AGCs, so that this data confirmed the guide lines proposed in this issue . The histologic examination of EGC confirmed the data of echoendoscopy about nodal status. CONCLUSIONS: Laparoscopic surgery is a safe and feasible procedure both for benign and for malignant lesions of the stomach. The results analysed hereby suggest that at the start of training be treated patients affected by benign lesions, followed by patients with EGC and then by patients with AGC. For gastric cancers, the average number of 18 nodes harvested from each patient was adequate, complying with the requirements suggested by the latest TNM classification. This choice of progressive selection of patients for training represents a good means to get an optimal performance level, especially in view of the oncologic requirements, and can prevent surgeons from elevated conversion rates and disappointing outcomes at the beginning of experience. PMID- 15902051 TI - Endoscopic biliary stenting facilitates safe and early removal of T-tube in liver transplant patients. AB - AIM: Duct to duct anastomosis in orthotopic liver transplant (OLT) patients have been traditionally performed with a t-tube in place for 3 to 6 months. Following removal of the t-tube a high incidence of biliary leakage has been reported. METHODS: Prospective study to evaluate the role of endoscopic biliary stenting to facilitate early and uncomplicated t-tube removal. All patients with duct to duct biliary anastomosis who had a t-tube in situ, from January 1998 to December 2002 were included in this study. RESULTS: There were 29 patients eligible for the study. Eight patients were not included due to early death. A protocol t-tube cholangiogram was performed in all patients, (median 12 days; range 4-47 days) followed by an endoscopic stent insertion (median 37 days; range 20-55 days). The stent was removed later (median 84 days; range 45-133 days). All complications related to the procedure were noted. Stent insertion was successful in all cases. In 2 patients a second endoscopic retrograde cholangiopancreatography (ERCP) was necessary, either because of failure to cannulate the papilla or to reposition the stent. There was a patient who presented a biliary leak due to stent displacement requiring a laparotomy. There were two further biliary leaks, one of them in an asymptomatic patient, which were managed conservatively. In addition 1 patient developed a mild case of postERCP pancreatitis. CONCLUSIONS: In liver transplants patients with an end-to-end choledochostomy with a t-tube, endoscopic biliary stenting allows an early removal of the T tube, with few complications. PMID- 15902052 TI - [Post-thyroidectomy cervical hematoma]. AB - AIM: Postoperative hematoma is a complications of thyroid surgery uncommon but potentially life threatening. It has implications for the trend toward outpatient procedures. METHODS: Retrospective review of 1.221 thyroidectomies performed at our institution over a 6-years period, to identify patients with hematomas requiring reoperation. Symptoms, treatment and findings at reoperation were evaluated. A control group (n=120) was compared for perioperative risk factors and outcome. RESULTS: Eighteen patients (1.5%) developed a postoperative hematoma. Symptoms included neck pain/pressure in 10 patients, respiratory distress in 9, wound drainage in 2, dysphagia in 1, agitation and sweating in 1. Mean time to symptom onset was 12 hours (range: 1.3-40 hours). Six hematomas presented between 7 and 24 hours, and 3 beyond 24 hours. Six patients required bedside hematoma evacuation. The bleeding source was identified in 15 patients. All patients recovered well, but one required a temporary tracheostomy. Case/controls comparison yielded in the study group a higher prevalence of hyperthyroidism (55.6% vs 25.8%, P=0.022) and intrathoracic goiter (50% vs 22.5%, P=0.029), and a longer mean hospital stay (5.22 vs 4.1, P=0.012); morbidity was not increased. CONCLUSIONS: Postoperative hematoma is an uncommon complication of thyroid surgery. If treated promptly, serious consequences can be avoided. The relatively long interval between the initial operation and the hematoma development needs to be considered when establishing outpatient practice guidelines. PMID- 15902053 TI - [Appendicectomy: laparoscopic or ''open approach''?]. AB - AIM: The aim of our study was to evaluate the advantages and disadvantages of open appendectomy versus laparoscopic appendectomy in the surgical treatment of acute appendicitis. We have compared the following items: operating time, postoperative pain, length of hospital stay, postoperative complications and costs. METHODS: The study was conducted on 435 patients admitted in our Department from December 1993 to December 2003 with diagnosis of acute appendicitis: 339 (77.9%) cases were operated with laparoscopic approach (LA group) and 96 (22.1%) cases with open approach (OA group) according to personal experience of surgeons on laparoscopic technique and patient's anthropometrical conformation. RESULTS: Mean operative time in LA group was 50 minutes (range 25 195) and 65 minutes (range 35-160) for OA group. In 15 patients (4.4%), the operation had to be converted to open approach. The morbidity was observed in 4.4% of patients for the LA group and 14.6% for the OA group. Hospital stay was faster for patients having laparoscopic appendectomy (2.5 days vs 3.5 days). Pain in the 1st and 2nd postoperative days, evaluated on the use of pain medication, was less in patient in LA group whereas the costs were higher in the LA group than in OA group. CONCLUSIONS: In our experience the laparoscopic approach to acute appendicitis can be considered safe and effective with diagnostic and therapeutic value. It significantly offers all the advantages of mini-invasive surgery reported in literature. PMID- 15902054 TI - [Appendicectomy: laparoscopic versus open treatment. A case survey]. AB - AIM: Laparoscopy is actually the gold standard approach in many surgical procedures: this consideration is still controversial as to appendectomy. METHODS: From 2000 to 2004 we have performed 257 appendectomies: 51 (20%) in laparoscopic approach. Preoperative diagnosis has been formulated on blood parameters, abdominal or, sometimes, transvaginal ultrasonography. RESULTS: Two hundred and fifty-seven surgical operations, 62 laparoscopic, have been performed for suspicious appendicitis. In the laparoscopic procedures, 11 revealed various diseases without appendicitis. In the remaining 51 cases, appendectomy has been performed totally intra-abdominal and none case turned to laparotomic conversion. Operative times were between 27 and 105 min in the laparoscopic appendectomies (LA) and between 18 and 46 min in the laparotomic appendectomies (OA). In 7.3% of OA and in 3.9% of LA wall infections occurred, as well as abdominal abscesses in 1% of OA and in 4% of LA. Postoperative discharge was after 3.3 days and bowel canalization appeared at 10-18 hours from the surgery, in OA and in LA. CONCLUSIONS: On the basis of these results, the conclusion is drawn that, although discordant opinions in the literature, the advantages of the laparoscopic approach compared to laparotomic approach is still to be demonstrated, both in advantages for the patient and in costs. Laparoscopy is the better surgical technique when the preoperative diagnosis is not clear, particularly in young women or in elderly, in whom a colic neoplasm may be suspected. PMID- 15902055 TI - [Verrucous carcinoma of the esophagus. A case report]. AB - Verrucous carcinoma (VC) of the esophagus is a rare variant of squamous cell carcinoma and only 20 cases have so far been reported in the international literature. The neoplasm is usually highly differentiated, presents a slow growth pattern with invasion of surrounding organs rather than blood-borne metastases. Recently, a causative role of human papillomavirus (HPV) has been hypothesized. The case of a patient affected with locally advanced VC of the esophagus and treated by means of local antiviral therapy is reported. A 41-year-old male patient was admitted to our institution for persistent atypical thoracic pain. The imaging techniques (thoracic and abdominal CT scans; upper GI endoscopy; hydrosoluble contrast swallow; endoscopic US) revealed a cauliflower-like protruding esophageal mass, active mucosal mycosis, multiple ulcerations of the distal esophagus, as well as 2 broncho-esophageal fistulas. The neoplasm extended beyond the esophageal wall, infiltrating surrounding cervical and mediastinal organs and the patient presented with secondary esophageal achalasia and right bilobar pneumonia. The histologic specimen was consistent with VC of the esophagus and the presence of HPV infection was detected by means of qualitative PCR assay. The patient was deemed not fit for surgery and a local antiviral treatment with hydroxy-phosphonyl-methoxypropyl-cytosine 5 mg/kg a week was started. After initial response to treatment, the patient presented with sudden progression leading to further broncho-esophageal fistula treated with endoscopic stent placement and ultimate death 6 months after referral to our center. In keeping with international data, our case confirms that esophageal VC has a highly unfavorable outcome, despite its high degree of differentiation and slow growth pattern. The long natural history, the lack of specific symptoms and the presence of coexisting esophageal diseases delay the diagnosis and account for the local advancement of this malignancy. Surgery is the option of choice for early stage lesions and advanced VC does not seem to benefit from current chemotherapeutic regimens. The causative role of HPV and the advancements of molecular pharmacology might allow for effective treatment in high-risk patients. PMID- 15902056 TI - Endometriosis in a trocar tract: is it really a rare condition? A case report. AB - In literature an elevated number of isolated cases of endometriosis in post laparoscopic scar or laparoscopic trocar tract are described. Actually no theory can completely account for endometriosis, and it is most likely that a combination of events is responsible for it. The case of a 37-year-old woman with a nodular mass in the right hypochondrium is reported. The nodule appeared after a laparoscopic cholecystectomy. Surgical excision was performed and microscopic analysis showed skeletal muscle and fibrous connective tissues with a typical glandular proliferation as in endometriosis. Endometrioma etiology is far to be cleared; the most practical and popular explanation is direct implantation. Our case may be explained according to this theory but it is difficult to achieve definitive conclusions due to the rarity of endometrioma and to the lack of information. PMID- 15902057 TI - [Surgical management of incidental Meckel's diverticulum: the necessity to obtain the informed consent]. AB - Surgical approach to the incidentally found Meckel's diverticulum (MD) is still source of debate: some support the systematic search and the surgical resection, others suggest to leave in situ the asymptomatic diverticulum. A wide literature review and some claims for malpractice referred to resection of diverticular formations (2 cases are described herein) allow us to deem that the removal of an asymptomatic MD may be an excess of surgical indication regarding an entity that cannot be defined as pathological for itself; anyway, the informed consent must be obtained before the incidental resection. The consent to the removal of MD must always be requested for laparotomies in which the surgeon forecasts the possibility to remove a diverticulum eventually found. Finally, an easy informed consent form about an incidentally found MD to be submitted to the patient is proposed. PMID- 15902058 TI - Periodontitis and preterm delivery. A review of the literature. AB - The aim of this paper was to review the relationship between periodontitis and preterm delivery (PTD). Original papers on this subject, published in English until the first quarter of 2004, were located in the MEDLINE/PubMed database. Additional papers were obtained by searching reference lists of previously published review papers. Case control studies pointed to an association between periodontitis and increased rates of PTD of low birth-weight (LBW) infants. Longitudinal studies showed that the incidence or progression of periodontitis during pregnancy may be an independent risk factor for PTD and LBW. Microbiological, immunological and animal model studies suggested that periodontal pathogens and their products may translocate to the fetal-placental unit resulting in PTD or fetal growth restriction. Maternal periodontal infection may also provide a chronic reservoir of inflammatory mediators and cytokines (TNF alpha, IL-1, IL-6, PGE2) that could adversely affect pregnancy outcome. Randomized controlled studies published indicated that periodontal treatment significantly reduces the risk of PTD and LBW infants. If these results are confirmed in further intervention studies, then prevention and treatment of periodontitis should be considered as a necessary part of prenatal care. PMID- 15902059 TI - In vitro effects of different concentration of PRP on primary bone and gingival cell lines. Preliminary results. AB - AIM: Platelet rich plasma (PRP) is utilized in oral surgery to enhance bone healing and it has been suggested to accelerate soft tissue healing. Nevertheless, there is no evidence on biological concentration of platelets needed to determine the higher biological response. Therefore, aim of this study is to evaluate the action of PRP in vitro on osteoblasts and fibroblasts and to evaluate which is the most effective concentration of PRP. METHODS: PRP was obtained from volunteers donors by standard apheresis. Osteoblasts were growth for 72 hours in a medium added with platelet concentration of 230%. Fibroblasts were treated with different platelet density for 24 hours and 72 hours. Platelet density was increased of 230%, 350%, 460% and 700% the normal blood count. Cell proliferation was evaluated with MTT test. ANOVA test was used to assess cells proliferation data. RESULTS: Osteoblasts proliferation, at 72 hours, showed an increase of proliferation in PRP group compared to plasma (P < 0.001). Fibroblast proliferation after 24 hours increases when PRP is added (P < 0.05). However, no significant differences were detected among the various concentration of PRP. Yet, at 72 hours, MTT values increases when platelet concentrate is 230% and 350%. At 700% platelet density MTT values were lower than control group (P < 0.05). CONCLUSIONS: Our results indicated that PRP has an enhancing effect on osteoblasts and fibroblasts proliferation when it is prepared only within certain ranges of concentrations. However, further experimental studies are needed to confirm our hypothesis. PMID- 15902060 TI - Counting of platelet derived growth factor and transforming growth factor-beta in platelet-rich-plasma used in jaw bone regeneration. AB - AIM: Growth factors (GFs) as platelet derived growth factor (PDGF) and transforming growth factor (TGF-beta), found in platelet beta-granules also present in platelet-rich-plasma (PRP), accelerate bone revascularization and regeneration and for this reason they have been employed successfully in dental and maxillofacial surgery. Platelet concentrate is commonly used for this purpose as long as platelet release reaction and the consequent GFs loss are avoided. To reduce this phenomenon we set up an easy and fast procedure for preparing a satisfying clotted PRP by adding CaCl2 only (no exogenous thrombin). METHODS: ELISA essay has been used to measure PDGF and TGF-beta in plasma, platelets and serum and platelet GMP-140, with the cytofluorometric technique in order to quantify the degranulation entity. RESULTS: In the 13 examined patients receiving clotted PRP to enhance bone regeneration in post-extractive alveolar sockets, PRP showed no sign of platelet activation (degranulation) and short recalcification times (8-12 min). The autologous clotted PRPs specimen have been evaluated in laboratory in terms of GFs percent: 76% of initial GFs content could be recovered in clotted PRP. This result confirms the absence of platelet degranulation in our procedure. CONCLUSIONS: Significant clinical results in alveolar bone regeneration are reached only with a high percentage of GFs inserted in bone matrix, avoiding early platelet degranulation. PMID- 15902061 TI - Reaction exothermia of 2 relining resins for temporary crowns. AB - AIM: The aim of the study was to evaluate the reaction exothermia of 2 relining resins for temporary crowns. The resins used were Duralay, a methacrylate-based resin, and Protemp II, a bisacrylic resin. METHODS: The coronal parts of an adult molar and of an impacted one were used. Abutments were obtained from the crowns, while the radicular part was sectioned in order to insert a thermocouple into the pulpal chamber. During the experiment the tooth with the polymerising resin was put into an oral cavity simulator to keep a constant temperature. The thermal increase during polymerisation of the 2 resins was recorded for 6 minutes at 5 seconds intervals and with 1 and 2.5 mm dentin thickness (n = 10). Data were statistically analysed by variance analysis with Scheffe test. RESULTS: The intrapulpal thermal increase was significantly higher in relation to the type of resin (Duralay p < 0.0001), thickness (1 mm p < 0.0001) and type of dentin (young dentin p < 00001). CONCLUSIONS: In order to protect the pulp, during the direct relining of a temporary crown, it is recommendable to choose the best combination among resin, type of dentin and its residual thickness, besides adequate cooling techniques. PMID- 15902062 TI - Debridement and local application of tetracycline in the management of persistent periodontitis. Clinical and microbiological results after 12 months. AB - AIM: The purpose of this study was to evaluate the clinical and microbiological efficacy of local delivery of tetracycline in persistent periodontal lesions after topical therapy. METHODS: A total of 44 bilateral pockets 4-5 mm deep and bleeding on probing were selected in 11 non-smokers patients, 22 treated by scaling and root planing plus tetracycline fibres (test sites) and 22 with scaling and root planing alone (control sites) in a split-mouth design. Bleeding on probing, probing depth and clinical attachment level were recorded at baseline, 6 and 12 months postoperatively. The prevalence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus and Treponema denticola was monitored by PCR at baseline, fibres removal, 6 and 12 months following treatment. RESULTS: The improvements in clinical parameters were greater in the tetracycline than in the control group. The reduction of bleeding on probing scores was 86.4% and 40.9%, the decrease of probing depth was 2.25 mm and 1.19 mm and the gain of clinical attachment level was 2.04 mm and 0.64 mm for test and control groups, respectively, over the 12 month period. The adjunctive use of tetracycline consistently resulted in a lower percentages of sites with detectable levels of A. actinomycetemcomitans, P. intermedia and P. gingivalis, while no differences were detected between treatments in the prevalence of T. denticola and B. forsythus. The pathogens could be eliminated from 3 periodontal pockets by debridement alone and from 12 sites by tetracycline at 12 months. CONCLUSIONS: Tetracycline local delivery gave the greatest advantage in the long-term treatment of periodontal persistent lesions. PMID- 15902063 TI - The dental needs of patients affected by infectious pathologies and/or seropositivity. AB - AIM: The aim of the study is to assess the demand for urgent dental treatment in patients carrying infectious pathologies and otherwise, particularly those with a history of drug addiction. METHODS: The Narni Hospital Unit of Odontostomatology has set up a dental clinic for patients suffering from infectious diseases. Considering the high number of emergency cases presenting, we developed a prospective research instrument to study the urgent problems of drug-addict patients carriers or otherwise of infectious viruses. Our study examined 4 years of activity, from 1997 to 2000, during which time 456 patients were examined, all former drug addicts. For each of them a basic protocol, including initial examination, investigation of patient history, any laboratory examinations necessary and, finally, conservative, demolition and prosthetic rehabilitative therapies, was carried out. RESULTS: Subjects referring to the dental clinic for urgent problems numbered 120, precisely 26% of the total population. Distinguishing patients on the basis of basic systemic pathology, we observed that 28% of infected patients and 21% of healthy patients requested urgent dental treatment. CONCLUSIONS: Former drug addict patients require an approach protocol, prophylaxis and specific, personalised therapy particularly where he or she is also a carrier of infectious pathologies. On the basis of our own personal experience, we describe how a good stomatological evaluation is necessary in this category of subjects who need dental care ever more frequently. PMID- 15902064 TI - Primary and exclusive gingival localization of non-Hodgkin lymphomas. Case report. AB - A case of primary and exclusive gingival localization of a non-Hodgkin lymphoma (NHL) personally observed at the Surgical Out-patients Unit of the Department of Dentistry at the Umberto I Hospital in Rome, is described. A 71 year-old caucasian male was referred by his private dentist because of a severe pain at the soft tissue of the mandible even after dental, neurologic and parodontal treatments. At clinical examination, a severe gingival swelling in the symphysis region with hard bleeding, palpable lymph nodes and no important evidence in radiographic and CT examinations were found. After incisional biopsy and its examination, a diagnosis of NHL was made. Further examinations did not show metastasis dissemination. The patient was referred to the Hematology Department and after 4 chemotherapy cycles over 12 months, he completely recovered. After an introduction on NHL, the clinical case is described as well as how to make a correct diagnosis; moreover, the importance of the dentist in identifying diseases even not strictly related to oral cavity but whose signs and manifestations appear over there, is stressed. PMID- 15902065 TI - Versatility of transmucosal implants with zirconium ring. Case report. AB - Utiling zirconium characteristics, a biomaterial with high affinity with bone and gum, we have positioned in the same patient 2 transmucosal implants in titanium with a coronal ring in zirconium in 2 different manners. The first fixture was positioned with the ring in zirconium at the same level of the gum in order to obtain a good periodontal healing and esthetic without metallic transparencies. The second postestractive implant was positioned with the zirconium ring at the level of the crestal bone in order to reduce the gap between the fixture and the alveolar bone and to avoid the use of a resorbable barrier relying on the osteointegrative properties of zirconium. PMID- 15902066 TI - Groove associated periodontitis: classification proposal and clinical. Case report. AB - The presence of a palatoradicular groove (PRG), more frequently observed at the level of the upper lateral incisors, may compromise both the radicular pulp and the periodontium leading to endodontic-periodontal lesions. Frequently, the prognosis of these elements is compromised because of the missed diagnosis or because of the complex therapeutic approach that is necessary to cure the associated pathologies. A histopathological and clinical classification is proposed showing according to the degree of the affected tooth involvement. Preventive measures are also discussed and a clinical case is reported. A 49-year old woman presented with a PRG at the superior left lateral incisor, endodontically treated years before, associated with probing pocket depths of 10 mm and 7 mm, clinical attachment loss of 11 mm and first degree mobility. The tooth was treated with splinting, periodontal surgery with radiculoplasty, root surface conditioning, guided tissue regeneration and with a prosthetic crown for esthetic rehabilitation. Six years later, the operated site showed probing pocket depths of 3 mm, gains in clinical attachment level of 4 mm and 7 mm and reduction of tooth mobility. The ceramic crown re-established an acceptable esthetics for the patient. Even in the presence of a 3(rd) degree PRG, according to the classification presented, the prognosis of the tooth may be positive through the association of root canal treatment, periodontal and prosthetic therapy. The classification presented shows the clinician the best therapeutic approach. PMID- 15902067 TI - Immediate loading of 4 implants. A case report. AB - Despite the obvious benefits of two-stage implants, patients do experience discomfort as they have to continue wearing non-fixed total dentures whilst postoperative healing takes place. These patients would be more comfortable if a functional and esthetically pleasing solution could be arrived at in a shorter time. In an effort to achieve this, the scientific community has researched and fitted ''immediate load'' implants. The current literature states that this procedure, when fitting 4 or more interforaminal implants, is both predictable and reliable. The procedure increases neither the number of implant failures nor the amount of bone lost. In the case reported, 4 implants were inserted and a bar constructed. The practicality of this procedure is seen in reduced time and costs. An initial total denture was used at the start, first as a surgery guide, then as an individual tray for impression, and eventually as final total denture. Fixing the initial total denture to the implant, considerably reduced the time taken in the whole implant procedure and provided the patient with a more comfortable, pleasing solution. The use of the bar prevented the denture from moving, guaranteed full support from the implants, and reduced or prevented the bone loss seen in patients where non-implant supported total dentures have been used. This procedure had the additional benefit of versatility - as it could have been used as an intermediate stage towards providing a more complex prosthesis, such as a Toronto prosthesis. PMID- 15902068 TI - Subantral filling by deantigenated heterologous bone and immediate fixture placement. AB - AIM: A successful implant therapy depends on sufficient quantity of available bone. Latero-posterior maxillary area often represents a hardly useful zone for routine implant therapy in long time edentulous patients. Standard procedures of maxillary sinus lifting are the goals of implant-prosthetic rehabilitation. Among different grafting materials used in this kind of procedures, the deantigenated heterologous bone reliability has been tested. METHODS: In a clinical case of maxillary bone atrophy (Misch classification, SA3) a deantigenated heterologous bone grafting and immediate fixture placement has been performed; before prosthetic finalization a biopsy in the grafting site has been carried out in order to obtain an histological evaluation of the graft integration. RESULTS: The histological analysis and X-ray at 1 year follow-up show the formation of osteoid tissue integrated with the surrounding bone tissue, reaching bone maturity by calcification. In spite of a longer time for grafting integration, the advantages of a single-time surgery and the reduction of surgical timing are stressed. CONCLUSIONS: X-rays and histological results lead authors to consider the tested heterologous bone as a valid alternative to autologous bone, especially in those patients where it is preferable to avoid a second surgical site in order to take an inlay graft. PMID- 15902069 TI - Disparities in deaths from stroke among persons aged <75 years--United States, 2002. AB - Despite declines in deaths from stroke, stroke remained the third leading cause of death in the United States in 2002, and age-adjusted death rates for stroke remained higher among blacks than whites. In 1997, excess deaths from stroke occurred among persons aged <65 years in most racial/ethnic minority groups, compared with whites. A younger age distribution among Hispanics and other racial/ethnic groups compared with whites might partly explain the disproportionate burden in deaths at younger ages. To examine disparities in stroke mortality among persons aged <75 years, CDC assessed several characteristics of mortality at younger ages by using death certificate data for 2002. This report summarizes the results of that assessment. Overall, 11.9% of all stroke deaths in 2002 occurred among persons aged <65 years; the proportion of stroke decedents who were aged <65 years was higher among blacks, American Indians/Alaska Natives, and Asians/Pacific Islanders, compared with whites. In addition, the mean ages of stroke decedents were statistically significantly lower in these racial groups than among whites. Blacks had more than twice the age-specific death rates from stroke than whites aged <75 years. Approximately 3,400 excess stroke deaths would not have occurred among blacks in 2002 if blacks had had the same death rates for stroke as whites aged <65 years. Moreover, age adjusted estimates of years of potential life lost (YPLL) before age 75 years from stroke were more than twice as high for blacks than for all other racial groups. Reducing premature death from stroke in these groups will require early prevention, detection, treatment, and control of risk factors for stroke in young and middle-aged adults. PMID- 15902070 TI - Regional and racial differences in prevalence of stroke--23 states and District of Columbia, 2003. AB - Higher stroke mortality in the United States has long been evidenced among blacks and residents of southeastern states. A greater proportion of blacks live in the southeastern states that make up the so-called stroke belt than elsewhere in the country; however, variations in socioeconomic characteristics and risk factors have also been associated with disparities in stroke, and these variations have been associated with region and race. To more closely examine these associations, CDC analyzed data from the 2003 Behavioral Risk Factor Surveillance System (BRFSS) survey. This report describes the results of that analysis, which indicated that although the prevalence of stroke was higher in 10 southeastern states than in 13 other states and the District of Columbia (DC) and higher among blacks than among whites, differences in education level and certain risk factors (i.e., having diabetes or high blood pressure, smoking, and not having health care coverage) might account for most of the differences in stroke prevalence by region and race. These findings reinforce the importance of primary and secondary prevention of known risk factors for stroke. PMID- 15902071 TI - Monitoring progress in arthritis management--United States and 25 states, 2003. AB - Arthritis is a chronic disease affecting an estimated 43 million (20.8%) U.S. adults and is the leading cause of disability in the United States. Arthritis results in activity and work limitations, decreased quality of life, and substantial burden to the U.S. health-care system. Promotion of arthritis self management through weight counseling, physical activity counseling, and arthritis education can reduce pain, improve function and quality of life, and delay disability among persons with arthritis. To encourage arthritis self-management, three objectives were added to the national health objectives for 2010. To monitor progress toward achieving these objectives and assess that progress by selected characteristics, CDC analyzed data from the 2003 National Health Interview Survey (NHIS) and the state-based 2003 Behavioral Risk Factor Surveillance System (BRFSS) survey. This report summarizes the results of those analyses, which indicated no statistically significant progress toward reaching the targets for weight counseling, physical activity counseling, and arthritis education. To meet these targets by 2010, public health and health-care agencies should increase efforts to improve awareness of these three factors among both health-care providers and patients. Such interventions will enable persons with arthritis to better self-manage their disease. PMID- 15902073 TI - Don't be shocked. PMID- 15902074 TI - It could happen to you. PMID- 15902075 TI - Follow the leader. PMID- 15902076 TI - The greatest volunteer of all. PMID- 15902078 TI - Potential legal nightmare. Simple case, complex resolution. PMID- 15902080 TI - Missed pericardial tamponade. When a medical patient becomes a trauma patient. PMID- 15902081 TI - A rational response to Taser strikes. PMID- 15902082 TI - How the Airway Report Form improved prehospital airway care and hospital communications. PMID- 15902083 TI - Improving not-for-profit EMS. Six self-sabotaging traps to avoid. PMID- 15902084 TI - On-scene, interagency conflict. Ways to keep agency differences from affecting patient care and your public image. PMID- 15902085 TI - Follow the cones. Highway scenes and your protection. PMID- 15902086 TI - Managing the pregnant trauma patient. PMID- 15902087 TI - Two new ways to get the doc on board. PMID- 15902090 TI - Touch them, then tech them. PMID- 15902089 TI - Pain-in-the-neck lawyers. PMID- 15902093 TI - Proteinuria or peptiduria: where's the beef? PMID- 15902094 TI - Foie gras. PMID- 15902095 TI - Decreased synthesis of glomerular adrenomedullin in patients with IgA nephropathy. AB - Adrenomedullin (AM) immunostaining and gene expression have seldom been measured in human kidneys. Because previous studies have shown that AM exerts antiproliferative effects on rat mesangial cells in vitro and that urine AM levels are decreased in patients with chronic glomerulonephritis, we measured glomerular AM and its gene expression in patients with primary IgA nephropathy (IgAN). Glomerular AM was measured by immunohistochemical staining, and glomerular AM mRNA was measured by in situ hybridization. Plasma and urine AM were measured by radioimmunoassay. The results showed that both the intensity of immunostaining for glomerular AM and the glomerular expression of AM mRNA were significantly decreased in IgAN patients compared with normal controls (both P < .05). Similar results were not observed in patients with non-IgA MsPGN. Glomerular AM immunostaining and glomerular AM mRNA expression were significantly correlated ( P < .001), and both were negatively correlated with the number of glomerular cells ( P < .05 and < .01, respectively). Both glomerular AM immunostaining and glomerular AM mRNA expression were correlated with urine AM levels (both P < .001), but not with plasma AM levels. The urine AM level was significantly lower in IgAN patients than in normal controls ( P < .01), whereas the plasma level was not different between the 2 groups. Our findings indicate that glomerular production of AM was decreased in patients with IgA nephropathy and that this lack of glomerular AM may be related to the pathogenesis of this mesangial disease. PMID- 15902096 TI - Urinary-peptide excretion by patients with and volunteers without diabetes. AB - Large quantities of peptides (1-4 g) are excreted in human urine each day. In this study we sought to analyze how peptide excretion varies with increasing albuminuria associated with diabetes, as well as to characterize the size distribution of albumin-derived peptides in urine from volunteers without diabetes and from patients with macroalbuminuria and diabetes. We detected albumin-derived peptides by injecting tritiated albumin intravenously into human volunteers and patients with diabetes. Urine was collected after 24 hours and fractionated on a size-exclusion column. This fractionation revealed peptides with molecular weights ranging from 300 to 500 Da in volunteers without diabetes. The albumin-derived peptides were of higher molecular weight in the urine of a patient with macroalbuminuria and diabetes. The molecular-weight distribution of the peptides derived from tritiated albumin peptides was paralleled by the distribution of all protein peptides (including albumin) as determined with the use of the Biuret protein assay or absorbance at 214 nm. We determined peptide excretion rates by filtering urine from patients with diabetes through a 10,000 Da molecular-weight-cutoff membrane and then measuring the filtrate with the use of the Biuret assay. This analysis revealed that the peptide-excretion rate increased with increasing total protein excretion, regardless of whether the patient demonstrated normoalbuminuria or microalbuminuria. Among patients with macroalbuminuria, the peptide-excretion rate leveled off and even decreased in the face of an increasing albumin concentration or protein-excretion rate. This study confirms that albumin-derived and protein-derived peptides exist at high concentrations in urine. Although peptide-excretion rates are maintained at similar levels up to macroalbuminuric states, the relative proportion of peptide excretion is significantly reduced compared with total protein. PMID- 15902097 TI - Increased susceptibility of fat-laden Zucker-rat hepatocytes to bile acid-induced oncotic necrosis: an in vitro model of steatocholestasis. AB - Metabolic liver disorders cause chronic liver disease and liver failure in childhood. Many of these disorders share the histologic features of steatosis and cholestasis, or steatocholestasis. In this study we sought to (1) develop an in vitro model of steatocholestasis, (2) determine the mechanisms of cell death in this model, and (3) determine the role of mitochondrial disturbances in this model. METHODS: Hepatocytes were isolated from 8-week-old obese (fa/fa) and lean Zucker rats. Cell suspensions were treated with glycochenodeoxycholic acid (GCDC), after which reactive oxygen species (ROS) generation, oncotic necrosis, apoptosis, and ATP content were assessed. Isolated liver mitochondria were exposed to GCDC and analyzed for ROS generation, mitochondrial membrane permeability transition (MPT), and cytochrome c release. Oncotic necrosis was significantly increased and apoptosis reduced in fa/fa hepatocytes exposed to GCDC compared with that in lean hepatocytes. Necrosis occurred by way of an ROS- and MPT-dependent pathway. Basal and dynamic ATP content did not differ between fa/fa and lean hepatocytes. GCDC stimulated ROS generation, MPT, and cytochrome c release to a similar extent in purified mitochondria from both fa/fa and lean rats. These findings suggest that fat-laden hepatocytes favor a necrotic rather than an apoptotic cell death when exposed to low concentrations of bile acids. The protective effects of antioxidants and MPT blockers suggest novel therapeutic strategies for the treatment of steatocholestatic metabolic liver diseases. PMID- 15902098 TI - Oligonucleotide-microarray analysis of peripheral-blood lymphocytes in severe asthma. AB - CD4 + lymphocytes play a key role in asthma pathogenesis, but much remains unknown about the genetic mechanisms that affect disease severity. In this study we sought to investigate global patterns of gene expression in CD4 + lymphocytes isolated from subjects with severe asthma through the use of microarray technology. CD4 + lymphocytes were separated from peripheral blood, total RNA was purified, and biotinylated complementary RNA was prepared and hybridized to Affymetrix HU133 chips (Affymetrix, Santa Clara, Calif). Using the robust multi chip average procedure, we compared the messenger RNA expression profiles of more than 33,000 genes of CD4 + lymphocytes in subjects with severe ( n = 5) and mild ( n = 5) asthma. Forty genes had 2-fold mean expression differences or greater. Thirty-seven genes were up-regulated, including transforming growth factor-beta and those involved in T-cell activation, proliferation, and cytoskeletal changes. Three genes were down-regulated, including the T-cell-receptor delta locus. This study demonstrates a method by which CD4 + lymphocytes can be extracted from blood for the purpose of microarray analysis. Furthermore, we show that T lymphocytes from the peripheral blood of subjects with severe and mild asthma differ in their gene-expression profiles, supporting the view that asthma is a systemic disease. These differentially expressed genes identify potential molecular targets for preventive and therapeutic options for severe asthma. PMID- 15902099 TI - Proteome of H-411E (liver) cells exposed to insulin and tumor necrosis factor alpha: analysis of proteins involved in insulin resistance. AB - Insulin resistance may be modeled in H-411E liver cells in tissue culture with the use of the cytokine tumor necrosis factor-alpha (TNF-alpha) and insulin. This tissue-culture model nicely mimics IR in human type 2 diabetes mellitus. After incubation of liver cells in tissue culture with INS alone, TNF-alpha alone, and TNF-alpha plus insulin, as well as a control sample, liver-cell extracts were separated on 2D polyacrylamide-gel electrophoresis on the basis of isoelectric point and molecular weight. We analyzed the gel images with the use of PD Quest software (Bio-Rad Laboratories, Hercules, Calif) to identify differentially expressed protein spots (ie, up or down with insulin vs down or up with TNF-alpha plus insulin). In separate experiments, phosphorus-32 incorporation/autoradiography and phosphoprotein staining were used to characterize treatment-induced phosphorylations. Affected protein spots were identified with the use of peptide fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry. The first series of experiments identified 6 differentially expressed proteins: eukaryotic translation initiation factor-3, subunit 2, regulator of G-protein signaling-5, superoxide dismutase, protein disulfide isomerase A6, proteasome subunit-alpha type 3, and regucalcin. In addition, we observed changes in the phosphorylation of protein disulfide isomerase A6. A second series of experiments identified 7 additional proteins with significantly altered differential expression: cell division protein kinase-4, kinogen heavy chain, carbonic anhydrase-7, E 3 ubiquitin protein ligase, URE-B1; Rab GDP dissociation inhibitor-beta, Rab GDP dissociation inhibitor-beta2, and MAWDBP. It can be seen that differentially expressed proteins, affected by treatment with insulin or with TNF-alpha plus insulin, include regulators of translation, protein degradation, cellular Ca ++ , G-proteins, and free-radical production. Although one cannot detail the mechanism or mechanisms of TNF-alpha induced IR from this data alone, it is easy to relate all of these proteins to a role in insulin signal transduction and, hence, insulin resistance. PMID- 15902107 TI - Clinical commentary: Report from the Society for Maternal-Fetal Medicine. PMID- 15902108 TI - Elevated first-trimester nuchal translucency increases the risk of congenital heart defects. AB - OBJECTIVE: We sought to evaluate the association between first trimester nuchal translucency measurement and the risk for major congenital heart defect in chromosomally normal fetuses. STUDY DESIGN: First trimester (10 weeks 4 days of gestation to 13 weeks 6 days of gestation) nuchal translucency was obtained in a large prospective multicenter National Institute of Child Health and Human Development study for Down syndrome prediction. The study, which was conducted between May 1998 and December 2000, was restricted to singleton pregnancies. Gestational age was determined by crown rump length measurements. Perinatal outcomes were determined and included the frequency of major congenital heart defect, which was defined as those cases that potentially could require surgery, intensive medical therapy, or prolonged follow-up time. Logistic regression analysis was used to determine whether nuchal translucency was a significant predictor of congenital heart defect. RESULTS: There were 8167 chromosomally normal pregnancies, of which 21 cases of major congenital heart defect were identified at follow-up examination (incidence, 2.6/1000 pregnancies). The risk of congenital heart defect rose with increasing nuchal translucency measurements. The mean nuchal translucency value for the normal and congenital heart defect groups were 1.5 mm and 1.9 mm, respectively (P = .05). With a nuchal translucency measurement of < 2.0 mm, the incidence of congenital heart defect was 13 of 6757 pregnancies (1.9 of every 1000 pregnancies). At 2.0 to 2.4 mm, the incidence was 5 of 1032 pregnancies (4.8 of every 1000 pregnancies). At 2.5 to 3.4 mm, the incidence was 2 of 335 pregnancies (6.0 of every 1000 pregnancies). At > or = 3.5 mm, the incidence was 1 of 43 pregnancies (23 of every 1000 pregnancies). Logistic regression analysis confirmed that nuchal translucency was associated significantly with congenital heart defect (odds ratio, 2.1; 95% CI, 1.4-3.1; P = .0004). CONCLUSION: Increased first trimester nuchal translucency measurement was associated with a higher risk of major congenital heart defect in chromosomally normal pregnancies. The practical implications of our findings are that patients with unexplained elevations of nuchal translucency may need referral for a fetal echocardiogram. PMID- 15902109 TI - Duration of human chorionic gonadotropin surveillance for partial hydatidiform moles. AB - OBJECTIVE: Partial hydatidiform moles infrequently progress to gestational trophoblastic neoplasia. The purpose of this study was to determine the optimal duration of human chorionic gonadotropin surveillance. STUDY DESIGN: We retrospectively reviewed the clinical follow-up of all women who were diagnosed with partial hydatidiform mole at our institution from 1983 to 2003. RESULTS: One hundred sixty-three patients were identified with a median age of 23 years (range, 14-42 years). Seventy-four patients (45%) attained undetectable levels of human chorionic gonadotropin; none of the patients had gestational trophoblastic neoplasia. Forty patients completed the 6 months of recommended follow-up; 6 patients conceived during surveillance, and 28 patients did not return for any further office visits 1 to 5 months after achieving remission. Eighty-three patients (51%) were lost to follow-up before normalization of human chorionic gonadotropin. Six women (4%) had stage I gestational trophoblastic neoplasia during surveillance. CONCLUSION: Our results support the suggestion that a single undetectable human chorionic gonadotropin level after evacuation is sufficient follow-up to ensure remission in patients with partial hydatidiform moles. PMID- 15902110 TI - Carboplatin and paclitaxel for the treatment of advanced or recurrent endometrial cancer. AB - OBJECTIVE: The purpose of this study was to determine the activity and toxicity of carboplatin and paclitaxel (taxol) in the treatment of advanced or recurrent endometrial cancer. STUDY DESIGN: This was a retrospective review of 18 consecutive patients with advanced (stage 4) or recurrent endometrial adenocarcinoma that had been treated with outpatient carboplatin and taxol. Taxol was delivered at 135 mg/m 2 over 3 hours, and carboplatin was delivery at an area under the curve of 5 over 1 hour. Cycles were repeated every 21 days. RESULTS: The overall response rate was 63% with 28% of patients who had a partial response and 35% of patients who had a complete response. Kaplan-Meier test was used to estimate the median survival time of 27 months and the median progression free survival time of 24 months. No patient had neutropenia, thrombocytopenia or grade 3 vomiting, neurosensory toxicity, or renal toxicity. CONCLUSION: Carboplatin and taxol for the treatment of advanced or recurrent endometrial cancer appear to be active regimens with minimal toxicity. PMID- 15902111 TI - The effect of hormonal contraception on the adequacy of colposcopic examination of the cervix. AB - OBJECTIVE: This study was undertaken to determine whether the use of hormonal contraception affects the adequacy of colposcopic examination of the cervix. STUDY DESIGN: A retrospective chart review of patients seen in the Vanderbilt University Medical Center Colposcopy Clinic from 2000-2002. Patient age, use of hormonal contraception, and adequacy of colposcopic examination were recorded for analysis. RESULTS: Of all the colposcopic records reviewed, 526 of 1156 met inclusion criteria. Study patients ranged in age from 14 to 51 years. The prevalence of the visible squamocolumnar junction ranged from 75% to 91%. The multivariate logistic regression analysis showed that both age and use of hormonal contraception significantly influence adequacy of colposcopic examination (P < .001, P = .001, respectively). CONCLUSION: The use of combination oral contraceptives is associated with an increased likelihood of adequate colposcopy. PMID- 15902112 TI - The role of sonographic endometrial patterns and endometrial thickness in the differential diagnosis of ectopic pregnancy. AB - OBJECTIVE: The purpose of this study was to examine the usefulness of the endometrial trilaminar pattern and thickness in the diagnosis of ectopic pregnancy. STUDY DESIGN: We reviewed patient records for clinical and ultrasonographic data for patients with the suspicion of ectopic pregnancy. The trilaminar pattern and endometrial thickness were tested as predictors for the diagnosis of ectopic pregnancy. RESULTS: The trilaminar pattern had a specificity of 94% and sensitivity of 38% (n = 403 women). The mean endometrial thickness was thinner in patients with ectopic, compared with normal pregnancy (9.5 +/- 5.7 mm vs 12.4 +/- 5.9 mm; P = .035). Patients with normal pregnancy or first-trimester losses had comparable thicknesses (12.4 +/- 5.9 mm vs 12.5 +/- 8.0 mm). The receiver operator curve showed that there was no thickness value useful for the diagnosis of ectopic pregnancy. CONCLUSION: The trilaminar pattern is specific for the diagnosis of ectopic pregnancy, but it is associated with low sensitivity. The endometrial thickness tends to be thinner in patients with an ectopic pregnancy. PMID- 15902113 TI - Predicting bacteriuria in urogynecology patients. AB - OBJECTIVE: This study was undertaken to determine whether reagent strip testing can predict bacteriuria in urogynecology patients. STUDY DESIGN: All women undergoing urodynamic evaluations from June 1997 to October 2001 were identified by using a computerized database. Urine culture results were compared with reagent strip testing. Significant bacteriuria was defined as greater than 10(5) colony-forming units per milliliter. RESULTS: Bacteriuria prevalence was 8.6% (n = 51). Sensitivity and specificity of nitrites were 0.51, (95% CI, 0.31-0.66) and 0.991, (95% CI, 0.974-0.998), respectively. Blood had a lower sensitivity (0.35, 95% CI, 0.20-0.54) and specificity (0.80, 95% CI, 0.75-0.84). Leukocyte esterase was similar to blood with a sensitivity of 0.28 (95% CI, 0.14-0.45) and specificity of 0.83 (95% CI, 0.78-0.87). No combination of tests offered improved sensitivity or specificity over nitrites alone. CONCLUSION: Nitrite dipstick testing has excellent specificity for bacteriuria in urogynecologic patients. These results support the treatment of women with positive nitrites who are preparing to undergo urodynamics without obtaining culture. PMID- 15902114 TI - Persistent and recurrent cervical dysplasia after loop electrosurgical excision procedure. AB - OBJECTIVE: Our objective was to determine whether demographic, colposcopic, and pathologic variables are predictive of recurrent cervical dysplasia. STUDY DESIGN: A retrospective review of patients who underwent loop electrosurgical excision procedure (LEEP) was performed. The medical records of the subjects were reviewed to identify demographic, pathologic, and procedural characteristics that predict recurrent dysplasia. RESULTS: A total of 514 subjects were identified who underwent LEEP between 1996 and 2003. Multivariate analysis revealed that advanced age, immunosuppression, and a positive endocervical margin were associated with recurrent dysplasia. CONCLUSION: Demographic and pathologic data can be used to predict the risk of recurrence of cervical dysplasia after LEEP. PMID- 15902115 TI - The effect of multiple cycles in oocyte donors. AB - OBJECTIVE: The purpose of this study was to determine whether multiple controlled ovarian hyperstimulation cycles in oocyte donors affect the ovarian response, the oocytes retrieved, or the pregnancy rates. STUDY DESIGN: A retrospective chart review of repeat donor in vitro fertilization cycles between 1992 and 2003 at the University of Cincinnati Center for Reproductive Health was performed. The variables that were examined included the peak estradiol level, the length of stimulation, the number of follicles >15 mm, the amount of gonadotropins that were used, the number of oocytes that were retrieved and inseminated, the average number of cells per embryo at the time of transfer, and the clinical pregnancy rates. RESULTS: A total of 107 in vitro fertilized donor oocyte cycles were analyzed, of which 45 young healthy women underwent at least 2 cycles and 17 women underwent 3 cycles. Donors who underwent a second or third cycle demonstrated no differences in the cycle parameters that were observed. CONCLUSION: Repeated controlled ovarian hyperstimulation cycles in a donor population does not demonstrate a diminished ovarian response to exogenous gonadotropins. Oocyte donors can undergo up to 3 stimulation cycles without a negative affect on the ovarian response to gonadotropins, the number of mature oocytes retrieved, the embryo quality, or the clinical pregnancy rate. PMID- 15902116 TI - Placement of a Word catheter: a resident training model. AB - OBJECTIVE: The present study developed and evaluated a practical teaching model and training protocol for Word catheter placement in the treatment of a Bartholin gland cyst. STUDY DESIGN: Residents were asked to treat a model of the female perineum, fashioned from harvested porcine tissue, with a simulated Bartholin cyst via Word catheter placement and drainage. Each resident was evaluated for procedural competence. They were retested after the viewing of a continuous 59 second video demonstrating catheter placement. RESULTS: The model's resemblance to female genitalia was confirmed by 68% of residents. Correct diagnosis and treatment of a Bartholin gland cyst was made by 61%. The mean pretraining score was 5.5 +/- 1.46, while the post-training average was 8.2 +/- 1.20, indicating an approximate 58% increase in resident proficiency (P < .05). CONCLUSION: A model of the female perineum fashioned from harvested porcine tissue is a suitable instructional aid in the recognition and treatment of a Bartholin cyst. Proficiency in Word catheter placement is improved through the use of the model and instructional video. PMID- 15902117 TI - Quality-of-life analysis in the management of endometrial cancer. AB - OBJECTIVE: The purpose of this study was to determine whether and how the quality of life (QL) of patients with stage I endometrial cancer was influenced by different surgical procedures with or without radiation therapy. STUDY DESIGN: We conducted a retrospective analysis of 200 women with stage I endometrial cancer at the University of Saskatchewan, Canada in 2001 through 2002. Modified QLQ-C30 Questionnaires were used in evaluating differences in the weighted QL of patients who underwent staged surgery and patients who had nonstaged surgery, the latter of which refers to total abdominal hysterectomy and bilateral salpingo oophorectomy (TAH-BSO) with or without radiation therapy. RESULTS: There was a significantly lower QL in patients who underwent staged surgery compared with nonstaged surgery. In addition, radiation therapy significantly worsened the QL of patients undergoing staged surgery, whereas it had little influence on the QL of patients who received nonstaged surgery. CONCLUSION: Our study suggests that nonstaged surgery with or without radiation therapy may be a preferred treatment for stage I endometrial cancer compared with staged surgery from the perspective of patients' QL. PMID- 15902118 TI - Predictors of contraception knowledge and use among postpartum adolescents in El Salvador. AB - OBJECTIVE: This study was undertaken to describe demographics and contraceptive familiarity and use among postpartum adolescents in El Salvador. STUDY DESIGN: Questionnaire-guided interviews were conducted in Spanish with 50 postpartum adolescents at an urban, public hospital in El Salvador. Open-ended questions included assessments of education, partnership status, and contraceptive knowledge and use patterns. RESULTS: The median age of subjects was 17 years, 84% were nulliparous, 80% had partners, and 6% were married. Eighty-four percent of the women reported contraception knowledge and 18% reported contraception use. Educational experience and literacy predicted contraceptive knowledge (P = .008 and .001, respectively), but not use. After delivery and postpartum contraception education, 58% of the subjects stated intention to use contraception. Having a partner and living with him were predictors of intent to use contraception (P = .001 and .002, respectively). Being single negatively predicted intention to use contraception (P = .001). CONCLUSION: Education and literacy predicted contraceptive knowledge; however, contraceptive knowledge did not predict contraceptive use. Adolescent contraception use depends on more than just contraceptive knowledge. PMID- 15902119 TI - Evaluation of the aged female baboon as a model of pelvic organ prolapse and pelvic reconstructive surgery. AB - OBJECTIVE: This study examines the baboon as an animal model of pelvic organ prolapse (POP) by describing the pelvic floor anatomy and adapting human clinical assessment tools. STUDY DESIGN: The pelvic anatomy of an adult female baboon was observed at necropsy, and comparisons were made to the human and squirrel monkey. The pelvic organ prolapse quantification (POP-Q) system was used to assess vaginal support in 12 living adult baboons, including 6 young, reproductive-age, nulligravid females (4.8 +/- 0.5 years) and 6 older, multiparous females (23.0 +/ 0.5 years). RESULTS: The female baboon pelvic anatomy was found to have similar architecture to the human and squirrel monkey female. Six multiparous females with mean parity of 5 (range 2-8) showed no evidence of POP or differences in POP Q measurements from 6 nulliparous females. CONCLUSION: The POP-Q system can be used to assess female baboon vaginal support. In a sample of baboons, pelvic support loss consistent with POP was not identified. As the pelvic anatomy is similar to the human female, the baboon may prove useful for evaluating surgical materials and for modeling pelvic floor reconstructive surgeries. PMID- 15902120 TI - Cellular response to chemotherapy and radiation in cervical cancer. AB - Effect of irradiation alone and irradiation after 5-fluorouracil (5-FU), paclitaxel, or cisplatin (CDDP) was investigated in human cervical cell lines (CaSki, ME180, SiHa, and C33A). High-risk human papillomavirus (HPV) (+) CaSki and SiHa cells were the most resistant to CDDP, 5-FU, and radiation treatments. Radiation and CDDP and 5-FU resulted in decreased survival of HPV 16 and 18 (+) cells, whereas addition of paclitaxel to radiation treatments decreased killing. Enhanced killing of ME180 cells containing HPV39 sequences was demonstrated with chemoradiotherapy with all agents. HPV(-) C33A was more sensitive to radiation than the other cell lines, and the addition of chemotherapeutic agents did not result in significant change in cytotoxicity. Expression of survivin was inversely proportional to cell sensitivity to CDDP, 5-FU, and radiation. Constitutive AKT levels are the lowest in cell lines that are the most resistant to CDDP, 5-FU, and radiation. These data provide correlation of response to combined therapeutic modalities with HPV status of cervical cancer and expression of survivin and AKT. PMID- 15902121 TI - Buccal misoprostol to prevent hemorrhage at cesarean delivery: a randomized study. AB - OBJECTIVE: The purpose of this study was to assess the efficacy of buccal misoprostol to decrease uterine atony, hemorrhage, and the need for additional uterotonic agents during cesarean delivery. STUDY DESIGN: Patients who underwent cesarean delivery were assigned randomly to either 200-microg misoprostol or placebo placed in the buccal space. A dilute intravenous oxytocin infusion was given to all patients at delivery of the placenta. The primary outcome variable was the need for additional uterotonic agents. RESULTS: A total of 352 women received random assignments. Demographic and intrapartum characteristics were similar between the groups. More women in the placebo group required 1 additional uterotonic agent (43% vs 26%; P < .01; relative risk, 1.3; 95% CI, 1.10, 1.50). There was not a difference between the groups in the incidence of postpartum hemorrhage or a difference in preoperative and postoperative hemoglobin level. CONCLUSION: Buccal misoprostol reduces the need for additional uterotonic agents during cesarean delivery. PMID- 15902122 TI - Clinical and sonographic estimation of fetal weight performed during labor by residents. AB - OBJECTIVE: This study was undertaken to assess the accuracy of both clinical and sonographic estimations of the fetal weight (EFW) performed during the active phase of labor by residents. METHODS: The study protocol consisted of achieving clinical, followed by sonographic EFW by the admitting resident during the active phase of labor. Patients who had an EFW (clinical or sonographic) within the last 2 weeks were excluded from the study. In addition, the effect of the following variables on the accuracy of the EFW were examined: maternal age, maternal weight and body mass index, parity, the Bishop score before obtaining the EFW, gestational age, birth weight, and the postgraduate year of the examiner. The Pearson correlation, chi2 test, and Student t test were the statistical analyses used. We also calculated the sensitivity, specificity, and positive and negative predictive values for clinical and sonographic EFW for detecting macrosomia (birth weight > or = 4000 g). RESULTS: A total of 192 patients participated in this study. The coefficient of correlation between the clinical and sonographic EFW and the actual birth weight was 0.59 (P < .0001) and 0.65 (P < .0001), respectively. Clinical EFW was correct (within +/-10%) in 72% of the cases and the sonographic EFW was correct (within +/-10%) in 74% of the cases. However, the sensitivity of predicting birth weight of 4 kg or more was only 50% for both clinical and sonographic EFW, with 95% and 97% specificity, respectively. None of the clinical variables that were tested were significantly associated with the accuracy of the EFW. CONCLUSION: Both clinical and sonographic EFW performed during the active phase of labor by residents correlate with the actual birth weight but have poor sensitivity in detecting macrosomic fetuses. Sonographic EFWs offer no advantage over clinical EFWs. PMID- 15902123 TI - Comparative longitudinal study of cervical length and induced shortening changes among singleton, twin, and triplet pregnancies. AB - OBJECTIVE: To compare cervical length and induced shortening changes during gestation among singleton, twin, and triplet pregnancies. STUDY DESIGN: Thirty two healthy gravidas (12 singleton, 13 twin, and 7 triplet pregnancies) between 17 and 20 weeks' gestation were prospectively enrolled in this longitudinal investigation of cervical length. Serial transperineal cervical length ultrasound assessments were made weekly until 34 weeks' gestation under 3 conditions: 1) supine, 2) supine with the Valsalva maneuver, and 3) standing. Cervical length, internal os diameter, and presence of cervical funneling were assessed under each condition. Multiple regression models were created using generalized estimating equations to predict these measures and accounting for confounding effects from covariates and adjusting for correlations from repeated measurements on each woman. RESULTS: A total of 1286 cervical sonographic measurements were made. In a multiple linear regression generalized estimating equations model, estimated cervical length was significantly different among singleton, twin, and triplet pregnancies. Overall, changing maternal position from supine to standing resulted in a nonsignificant change (-0.1 +/- 0.4 mm) in cervical length (P = .85). In contrast, measurement of cervical length during the Valsalva maneuver resulted in a significant reduction in cervical length when compared with the cervical length measured in supine (-1.0 +/- 0.3 mm) (P = .0009) and standing positions (-0.9 +/- 0.4) (P = .009). The observed induced shortening changes were similar across gestation, irrespective of singleton, twin, or triplet gestation. CONCLUSION: Longitudinal cervical length changes differ significantly throughout gestation among singleton, twin, and triplet pregnancies. Cervical length measurements made in a standing position are comparable with those measured while supine, whereas cervical lengths measured during the Valsalva maneuver are significantly shorter than those made in either the supine or standing position. PMID- 15902124 TI - Acute fatty liver of pregnancy in 3 tertiary care centers. AB - OBJECTIVE: The purpose of this study was to determine the demographics, clinical presentation, and maternal and neonatal outcomes in patients who were diagnosed with acute fatty liver of pregnancy over a 10-year period. STUDY DESIGN: This was a multicenter retrospective study of women with the diagnosis of acute fatty liver of pregnancy. Records were reviewed for symptoms, laboratory findings, clinical course, and maternal and perinatal outcomes. RESULTS: Sixteen cases of acute fatty liver of pregnancy were identified. Three of the 16 cases had multiple gestations (18%). Eleven of the 16 cases were diagnosed in the antepartum period, and 5 cases were diagnosed within 4 days after delivery. Nausea and vomiting were the most common symptoms (75%). There were 2 maternal deaths (12.5%) and 3 fetal deaths (15%). CONCLUSION: We recommend that patients with persistent nausea, vomiting, or epigastric pain in the third trimester receive evaluation of liver enzymes, renal function, and a complete blood count to rule out the diagnosis of acute fatty liver of pregnancy. PMID- 15902125 TI - Intertwin time interval: how it affects the immediate neonatal outcome of the second twin. AB - OBJECTIVE: Customarily 30 minutes is used as the upper limit of intertwin birth times. This study was conducted to define factors that influence the immediate neonatal outcome of the second twin and to assess whether a valid cutoff time is evident. STUDY DESIGN: We evaluated all live born viable vaginal twin deliveries (144) between November 1997 and December 2003. The relationship between intertwin birth time and neonatal outcome was then evaluated continuously and in 15 to 30 minute intervals. RESULTS: In multivariate analysis, only intertwin birth time predicted umbilical cord pH. Arterial/venous pH decreased linearly at a rate of 0.00063 and 0.00081 U/min. Venous pH was significantly lower in later 15-minute intervals compared with earlier ones (P = .002), and in patients delivering before or after a 30-minute cutoff (7.31 vs 7.26, P = .012). CONCLUSION: Our study confirmed that longer intertwin birth time is associated with a continuous slow decline in umbilical cord pH. However, the small differences in pH were not large enough to impact clinical management. PMID- 15902126 TI - Obesity as a risk factor for failed trial of labor in patients with previous cesarean delivery. AB - OBJECTIVE: This study was undertaken to determine whether obesity is a risk factor for failed trial of labor (TOL) in women with previous cesarean delivery (CD). STUDY DESIGN: We performed a review of singleton gestations 36 weeks or greater with previous CD who underwent TOL from January 1998 to June 2002, stratifying by body mass index (BMI, kg/m2): normal (BMI <25), overweight (BMI 25 29.9), obese (BMI 30-39.9), and morbidly obese (BMI >40). Rates for failed TOL were determined, and groups compared. RESULTS: For 725 patients, failed TOL rates were as follows: 14.1%, 20.4%, 27.7%, and 30.3% for normal, overweight, obese, and morbidly obese groups, respectively (P < .0001). Significant risk factors included: no previous vaginal delivery, labor induction, recurrent CD indication, and fetal macrosomia. However, obesity remained an independent risk factor for failed TOL in the obese and morbidly obese groups with odds ratio of 1.99 (95% CI 1.20-3.30) and 2.22 (1.11-4.44) for these groups (P = .03), respectively. CONCLUSION: Obesity is an independent risk factor for failed TOL in patients with previous CD. PMID- 15902127 TI - Fetal "nonreassuring status" is associated with elevation of nucleated red blood cell counts and interleukin-6. AB - OBJECTIVE: Previous studies have established the association between fetal hypoxia and elevated nucleated red blood cells (NRBCs). Animal studies have demonstrated that a rise in plasma erythropoietin (EPO) is not detectable until 4 to 6 hours after the initiation of hypoxia. In contrast, interleukin-6 (IL-6) has the capacity to directly induce erythroid maturation. Therefore, we set forth to evaluate the role of EPO and IL-6 as potential mediators of elevated fetal NRBCs in response to acute hypoxia. STUDY DESIGN: Low-risk pregnancies with a normal fetal heart rate at admission to labor and delivery were eligible for participation. Deliveries for "nonreassuring fetal status" comprised the study group. All other deliveries served as controls. Umbilical cord blood was prospectively collected for blood gas analysis, NRBC counts, EPO, and IL-6. RESULTS: One hundred women participated in the study. Nonparametric univariate analysis demonstrated a significant association between elevated NRBC counts and Apgar scores, arterial cord blood pH, base excess, EPO, and IL-6 levels (all P values <.01). Stepwise regression analysis identified only pH, IL-6, and EPO as independent variables associated with elevated NRBC counts at birth (all P values <.0001 with R2 of 0.27, 0.42, and 0.46, respectively). A significant increase in NRBC counts was noted in study patients. IL-6 was significantly increased in study patients, whereas there was no difference in EPO between groups. CONCLUSION: The fact that NRBC counts were elevated in fetuses who were delivered for "nonreassuring fetal status" with EPO being normal and IL-6 being elevated implies that IL-6 may have a unique, short-term role in elevating fetal NRBC counts. PMID- 15902128 TI - Magnesium sulfate tocolysis and pulmonary edema: the drug or the vehicle? AB - OBJECTIVES: This study was undertaken to determine the: (1) risk factors for developing pulmonary edema associated with magnesium sulfate (MgSO4) tocolysis; (2) mean latency period to diagnosis; (3) role of maternal transport; and (4) safety of continued therapy. STUDY DESIGN: A total of 150 antenatal patients treated for preterm labor with MgSO4 were identified for this retrospective, case control study. Cases were compared 1:2 with controls in regard to maternal demographics, MgSO4 concentration and infusion rates, maternal transport status, and maternal net fluid balance. RESULTS: Risk factors for developing pulmonary edema include: greater MgSO4 and intravenous fluid infusion rates, less concentrated MgSO4, infection, multiple gestations, concomitant tocolytics, large positive net fluid balances, and maternal transport. The mean latency period to diagnosis was 1.96 days. Six percent of patients had recurrence if MgSO4 tocolysis was continued. CONCLUSIONS: MgSO4 and intravenous fluid rates are both associated with the development of pulmonary edema. Once appropriately treated, MgSO4 tocolysis can be continued with little risk of recurrence. PMID- 15902129 TI - Neonatal impact of elective repeat cesarean delivery at term: a comment on patient choice cesarean delivery. AB - OBJECTIVE: The object of this study was to compare neonatal outcomes of term uncomplicated pregnancies delivered by scheduled repeat cesarean with outcomes of babies born to mothers intending to deliver vaginally. STUDY DESIGN: This retrospective cohort study describes neonatal outcomes of term uncomplicated pregnancies. Neonates of mothers intending to deliver vaginally (n = 3134) are compared with neonates born by elective repeat cesarean delivery prior to labor (n = 117). RESULTS: Neonates born by elective repeat cesarean are more frequently admitted to advanced care nurseries than infants born to mothers intending to deliver vaginally (risk ratio 3.58, 95% confidence interval 3.35-3.58). CONCLUSION: The decision to undergo scheduled cesarean delivery appears to negatively impact immediate neonatal outcomes. PMID- 15902130 TI - Comparison of early-onset neonatal sepsis caused by Escherichia coli and group B Streptococcus. AB - OBJECTIVE: The purpose of this study was to compare maternal characteristics and neonatal morbidity and mortality rates that are associated with early-onset neonatal sepsis that is caused by group B Streptococcus and Escherichia coli. STUDY DESIGN: This was a retrospective review of newborn infants with a positive blood culture (and/or cerebrospinal fluid) that was positive for either E coli or group B Streptococcus during the first week of life. Data were abstracted from maternal and neonatal medical records. RESULTS: Among 28,659 deliveries during the study period, 102 episodes of early-onset neonatal sepsis were identified, 61 of which were caused by group B Streptococcus and 41 of which were caused by E coli. E coli sepsis cases had a lower birth weight, a higher percentage with 5 minute Apgar score <7, and a longer stay in the hospital neonatal intensive care unit and required mechanical ventilation more frequently. Death after early-onset neonatal sepsis with E coli was also more frequent. CONCLUSION: Early-onset sepsis with E coli is associated with more morbidity and a higher mortality rate compared with early-onset group B Streptococcus. PMID- 15902131 TI - Modifiable risk factors for growth restriction in twin pregnancies. AB - OBJECTIVE: This study was undertaken to evaluate modifiable risk factors for adverse fetal growth in twin pregnancies. STUDY DESIGN: A large cohort study from a database of women with twin gestations identified at risk for preterm labor was performed. Examining each infant's birth weight and gestational age at delivery, infants were classified as being average (AGA), large (LGA), or small (SGA) for gestational age, using the Alexander reference curve. Clinical and demographic factors were compared between patients delivering at least 1 SGA infant and AGA pairs using Pearson's chi2 Student t test statistics and logistic regression. RESULTS: There were 11,827 twin pregnancies evaluated. Risk factors associated with SGA deliveries included tobacco abuse, poor weight gain, lean prepregnancy body mass index, African American race, and nonmarried. The logistic regression identified tobacco abuse as the single greatest risk for poor fetal growth, (odds ratio [OR] 1.95; 95% CI [1.68, 2.27]). Weight gain of less than one-half lb/wk also increased SGA risk (OR 1.35; 95% CI [1.16, 1.68]), whereas weight gain greater than 1 lb/wk decreased SGA risk (OR 0.77; 95% CI [0.68, 0.86]). CONCLUSION: Tobacco abuse and weight gain are the modifiable risk factors, which require intervention during a twin pregnancy. Patients should be encouraged to stop tobacco abuse and gain a minimum of one-half lb/wk in the later half of pregnancy to minimize the risk for growth restriction. PMID- 15902132 TI - The effect of short-term indomethacin therapy on amniotic fluid volume. AB - OBJECTIVE: The purpose of this study was to determine the effect of short-term (48 hour) indomethacin therapy on amniotic fluid volume. STUDY DESIGN: Patients who were admitted to Northwestern Memorial Hospital in preterm labor and who received indomethacin tocolysis were eligible for the study. Thirty-five patients with singleton gestations and 26 patients with twin gestations were enrolled. Amniotic fluid volumes were measured at baseline, at 48 hours after the initiation of indomethacin, and then every 24 hours thereafter for a maximum of 7 days. RESULTS: There were no significant changes in mean amniotic fluid volume subsequent to the administration of indomethacin at any of the time points that were measured. Among the 61 patients who received indomethacin, there were 2 cases of oligohydramnios (3.3%; 95% CI, 0.03%, 11.3%). In both cases, the amniotic fluid index returned to normal within 24 hours after the discontinuation of indomethacin. CONCLUSION: Short-term indomethacin therapy, in either singleton or twin gestations, does not cause a clinically significant change in amniotic fluid volume. PMID- 15902133 TI - Early intervention for perinatal depression. AB - OBJECTIVE: This study was undertaken to design a process that effectively identifies and facilitates early intervention for women in an obstetrics clinic who are at risk for postpartum depression. STUDY DESIGN: Under this new program, labeled ISIS (Identify, Screen, Intervene, Support), we educated our new obstetric patients and clinic staff about postpartum depression through patient education classes, departmental lectures, and handouts. Then, we implemented simple procedures to identify risk factors for depression at intake and screened for depressive symptoms at the 32-week visit using the Edinburgh Postnatal Depression Scale (EPDS). In addition, we facilitated treatment of at-risk or symptomatic patients with the introduction of a social work consultant in the clinic setting. RESULTS: In an obstetric chart review, 75% of our patients were screened for depression in pregnancy. Ten percent of these women demonstrated symptoms of depression warranting further evaluation. CONCLUSION: Preliminary data from our multidisciplinary approach suggest that educating, screening, and appropriately treating or referring these women can take place in a busy obstetric clinic. PMID- 15902134 TI - Management of cervical adenocarcinoma in situ during pregnancy. AB - OBJECTIVE: Adenocarcinoma in situ (AIS) is a precursor of invasive disease that is being more frequently diagnosed during the reproductive years. Few reports have described the treatment of this condition in gravid women. The purpose of this study was to review our collective experience managing cervical AIS during pregnancy. STUDY DESIGN: Retrospective medical record review of all women diagnosed with AIS during pregnancy from 1995 to 2004 at 3 academic institutions. RESULTS: Eleven women with a median age of 32 years were identified. Five who received a diagnosis in the early second trimester underwent uncomplicated cold knife conization (CKC) at 14 to 19 weeks' gestation. Six patients underwent postpartum CKC. All 11 women delivered at term. One patient undergoing postpartum CKC required radical hysterectomy for stage IB1 cervical adenocarcinoma. Four subsequent pregnancies occurred among patients having fertility-sparing surgery. CONCLUSION: Management of cervical AIS during pregnancy by early second trimester CKC is safe for mother and fetus. PMID- 15902135 TI - Effect of delivery route on natural history of cervical dysplasia. AB - OBJECTIVE: This study was undertaken to determine the (1) impact of delivery route on the natural history of cervical dysplasia and (2) overall regression rates of cervical dysplasia in pregnant women. STUDY DESIGN: A retrospective analysis was performed on 705 pregnant women with abnormal Papanicolaou tests who presented for prenatal care at the Kapiolani Medical Center Women's Clinic in Honolulu, Hawaii, between 1991 and 2001. Data collection included demographics, delivery route, and cervical pathology. RESULTS: Two hundred one patients met the inclusion criteria. Regression rates for vaginal and cesarean section groups were as follows: atypical squamous cells (64% vs 70%, P = .32), low-grade squamous intraepithelial lesion (58% vs 42%, P = .073), and high-grade squamous intraepithelial lesion (53% vs 25%, P = .44). Of the total population, 30% of lesions persisted postpartum, 58% regressed, and 12% progressed. CONCLUSION: Mode of delivery does not influence the natural history of dysplastic lesions. Gravid and nongravid women have similar regression rates. PMID- 15902136 TI - Postpartum follow-up of a positive purified protein derivative (PPD) among an indigent population. AB - OBJECTIVE: This study was undertaken to investigate the rates and predictors of follow-up and treatment for postpartum patients with a positive purified protein derivative (PPD). STUDY DESIGN: Retrospective cohort study of all women delivered at San Francisco General Hospital in 2000. All patients with a positive PPD were identified and their demographic and PPD follow-up and treatment data were collected and analyzed. RESULTS: Among the 1331 patients delivered, the prevalence of a positive PPD was 32% (n = 425). Of the 393 patients who had not been previously treated, 42% (n = 167) attended a follow-up visit with 42% of these (n = 71) actually completing 6 months of therapy. Among different ethnicities, Asian patients were more likely to follow-up at a rate of 52% (P = .03). Patients who received care from the same physician both antepartum and postpartum were more likely to attend and complete therapy at rates of 67% (P < .001) and 62% (P = .01), respectively. CONCLUSION: We found that despite the opportunity given by the interaction with the medical system during pregnancy, only 18% of patients with a positive PPD actually completed therapy. PMID- 15902137 TI - Abnormal placentation: twenty-year analysis. AB - OBJECTIVE: This study was undertaken to determine whether the rate of abnormal placentation is increasing in conjunction with the cesarean rate and to evaluate incidence, risk factors, and outcomes. STUDY DESIGN: Cases from 1982-2002 were identified by histopathologic or strong clinical criteria. Risk factors were assessed in a matched case-control study, and analyzed using conditional logistic regression models. RESULTS: There were 64,359 deliveries, with cesarean rates increasing from 12.5% (1982) to 23.5% (2002). The overall incidence of placenta accreta was 1 in 533. Significant risk factors for placenta accreta in our final analysis included advancing maternal age (odds ratio [OR] 1.13, 95% CI 1.089 1.194, P < .0001), 2 or more cesarean deliveries (OR 8.6, 95% CI 3.536-21.078, P < .0001), and previa (OR 51.4, 95% CI: 10.646-248.390, P < .0001). CONCLUSION: The rate of placenta accreta increased in conjunction with cesarean deliveries; the most important risk factors were previous cesarean delivery, previa, and advanced maternal age. PMID- 15902138 TI - The effect of the lunar cycle on frequency of births and birth complications. AB - OBJECTIVE: The purpose of this study was to examine the influence of the lunar cycle on the frequency of deliveries and/or delivery complications. STUDY DESIGN: This was a retrospective cohort, secondary analysis of 564,039 births across 62 lunar cycles that were identified from North Carolina birth certificate data from 1997 to 2001. RESULTS: Using analysis of variance and t-tests, we found no significant differences in the frequency of births, route of delivery, births to multigravid women, or birth complications across the 8 phases of the moon or between documented high- and low-volume intervals of the lunar cycle. CONCLUSION: An analysis of 5 years of data demonstrated no predictable influence of the lunar cycle on deliveries or complications. As expected, this pervasive myth is not evidence based. PMID- 15902139 TI - The effect of parenthood on perceived quality of life in teens. AB - OBJECTIVE: To describe the perceived quality of life of teen mothers. STUDY DESIGN: The Medical Outcomes Survey-Short Form 36, version 2 is a scale that measures a subject's perception of 8 health dimensions. The Medical Outcomes Survey-Short Form 36, version 2 and a demographics survey were completed by women during obstetric or gynecologic visits to a resident continuity clinic. Mean scores were compared between women with children and those without. RESULTS: There was no significant difference between adults or teens, with or without children, in any health component scale with the exception of social functioning. When compared with the normative population age, all groups in our population scored significantly on physical functioning and role-physical subscales. In addition, teens with children scored lower on the role-emotional subscale. CONCLUSION: Perceived quality of life in teen mothers does not appear to be lower than quality of life in teens without children or adult women. PMID- 15902140 TI - Prenatal diagnosis of 47,XXX. AB - We report 2 cases of 47,XXX that were diagnosed prenatally and were screened positive for trisomy 21 by biochemical and ultrasound markers. These cases underline the importance of discussing the sex chromosome abnormalities during the genetic counseling after an abnormal triple screen test or ultrasound examination. PMID- 15902141 TI - Does an obese prepregnancy body mass index influence outcome in pregnancies complicated by mild gestational hypertension remote from term? AB - OBJECTIVES: This study was undertaken to determine the influence of increased prepregnancy body mass index (BMI) on pregnancy outcome in women with mild gestational hypertension remote from term. STUDY DESIGN: A matched cohort design was used. A total of 365 pregnant women with mild gestational hypertension and a normal prepregnancy BMI (20-25 kg/m2) were matched 1:1 for gestational age at diagnosis, race and parity to 365 women with mild gestational hypertension, but an obese prepregnancy BMI (> or = 30 kg/m2). RESULTS: Matched characteristics were distributed as follows: gestational age at diagnosis 32.2 +/- 2.4 weeks; race 80.8% white; parity 59.7% nulliparous. The rate of progression to preeclampsia was the primary endpoint of the study and was similar between the prepregnancy normal and obese BMI groups (37.8% vs 41.1%, P = .352). Birth weight at delivery (3033 +/- 747 g vs 2833 +/- 659 g, P < .001) and cesarean delivery rate (56.7% vs 40.3%, P < .001) were greater in the obese prepregnancy BMI group. CONCLUSION: In patients with mild gestational hypertension remote from term, an obese prepregnancy BMI was associated with similar rates of progression to preeclampsia. Secondarily, an obese prepregnancy BMI was associated with higher birth weights and an increased incidence of cesarean delivery. PMID- 15902142 TI - A review of intrapartum fetal deaths, 1982 to 2002. AB - This study identifies cases of unexpected intrapartum fetal deaths over 20 years in a Canadian tertiary hospital. Of 121,659 births, 82 were intrapartum deaths. Eleven fetuses were considered viable and nonanomalous. Six deaths were deemed ideally preventable. Application of electronic fetal heart rate monitoring and rapid operative delivery may reduce the already low rate of intrapartum fetal deaths. PMID- 15902143 TI - Required research by medical students and their choice of a women's health care residency. AB - OBJECTIVE: This study searches for association between the subject of medical students' required research projects and subsequent choices of residency, focusing specifically on women's health fields. STUDY DESIGN: Students at our school graduating between 1997 and 2004 (n = 535) were required to undertake a research project beginning in the preclinical years. The subject of their required research project was related to their subsequent choice of residency. RESULTS: Ninety-nine of the 535 projects (18.5%) were women's health related. Although overall there was no significant relationship between the research project and residency choice, students who completed a women's health care project were nearly twice (1.8, 95% CI, 1.4-2.3) as likely to enter a women's health (obstetrics and gynecology or family medicine) residency than those who did not. CONCLUSION: An association exists between student participation in women's health research beginning during preclinical years and the subsequent choice of a women's health residency. PMID- 15902146 TI - Imperatives. PMID- 15902147 TI - The hidden epidemic of pelvic floor dysfunction: achievable goals for improved prevention and treatment. AB - Each year, pelvic floor dysfunction affects between 300,000 and 400,000 American women so severely that they require surgery. Approximately 30% of the operations performed are re-operations. The high prevalence of this problem indicates the need for preventive strategies, and the common occurrence of re-operation indicates the need for treatment improvement. Efforts at prevention and treatment improvement will only be possible if research clarifies causative mechanisms and scientifically valid studies discover why operations fail. By reaching a goal of 25% prevention we could save 90,000 women from experiencing pelvic floor dysfunction and with 25% treatment improvement we could avoid 30,000 women from needing a second operation. To achieve these goals we must discover specific events or behaviors in a woman's life that lead to these problems and that are amenable to preventive strategies. In addition, we must define specific biologic and behavioral factors that explain why certain women have recurrence after surgery. Because the pelvic organ support system is comprised of muscles, ligaments, and nerves arranged in a complex tension-based apparatus, the basic nature of this work must include biomechanical analyses of the overall mechanism and targeted research into the biology of muscle, ligament, nerve, and their complex interactions in normal pelvic floor function and in symptomatic patient. Each of these scientific disciplines is well developed so that engaging scientists in the effort to move forward will bring predictably important results. With an integrated approach to this problem over the next 20 years, it should be possible to achieve these goals and reduce the suffering for more than 100,000 afflicted women. PMID- 15902148 TI - Abdominal sacral colpopexy with allograft fascia lata: one-year outcomes. AB - OBJECTIVE: The purpose of this study was to assess 1-year outcomes of sacral colpopexy with the use of allograft fascia lata. STUDY DESIGN: Records of all subjects who underwent sacral colpopexy with allograft fascia lata from May 1, 2001, to April 30, 2003, were reviewed. Subjects underwent pre- and postoperative evaluation of prolapse with the pelvic organ prolapse quantification system. The Fisher's exact test was used to analyze the results. RESULTS: Allograft fascia lata was used for 24 colpopexies during this period. No significant intraoperative or postoperative complications or graft erosions occurred. Five subjects were lost to follow-up after 3 months. Analysis was performed on the remaining 19 subjects. Prolapse of stage 2 or more in compartments Aa, Ba, Ap, Bp, and C was preoperatively 50%, 74%, 78%, 84%, and 68% and postoperatively 11%, 16%, 21%, 26%, and 5%, respectively. CONCLUSION: Allograft fascia lata may be a suitable alternative to permanent mesh for sacral colpopexy, but longer-term outcomes and larger studies are needed. PMID- 15902149 TI - Vascular anatomy of the presacral space: a fresh tissue cadaver dissection. AB - OBJECTIVE: To assess variability in the vascular structures of the presacral space and to estimate the risk of injury because of blind suture placement during sacral colpopexy. STUDY DESIGN: Ten fresh frozen female cadavers were evaluated. Three 0-polyester sutures were placed blindly through the peritoneum and around the midline of the anterior longitudinal ligament. The presacral space was dissected and the sutures examined for injury to vessels. The midline of the anterior longitudinal ligament was marked from the promontory to its inferior edge, and measurements were taken to the leading edge of vessels proximal to the presacral space. On a template, all vessels larger than 2 mm were drawn to scale and overlaid on the template. RESULTS: Unequivocal vascular injury was found in 5 cadavers because of blind sutures. Four injuries occurred to the middle sacral artery and 1 to the left common iliac vein. There was significant variability in location of vessels, particularly on the left side of the ligament. CONCLUSIONS: The vascular pattern of the presacral space is variable, and major vessels may deviate significantly from their expected positions. Surgeons should carefully expose this space prior to placing sutures during sacral colpopexy. PMID- 15902150 TI - Immunohistochemical evidence for the interaction between levator ani and pudendal motor neurons in the coordination of pelvic floor and visceral activity in the squirrel monkey. AB - OBJECTIVE: The purpose of this study was to characterize the spinal distribution of afferent and efferent pathways that innervate the levator ani (LA) muscle in the female squirrel monkey. STUDY DESIGN: Cholera toxin B (CTB) was injected unilaterally into the LA muscle of 5 monkeys to identify primary sensory neurons in the dorsal root ganglia (DRG) and motor neurons in the spinal cord that contribute fibers to the LA nerve. Fluoro-Gold (FG) was injected into the external anal sphincter of 2 of these animals to label pudendal motor neurons (1 of these animals underwent unilateral LA neurectomy before CTB injection). Spinal cord and DRG were processed for immunofluorescence 3 to 7 days after injections. RESULTS: Retrograde transport of CTB from the LA muscle labeled primary afferent neurons in the ipsilateral DRG, their central projections, and motor neurons in the medial portion of the ipsilateral ventral horn of the spinal cord (L7-S2 segments). Injection of FG into the external anal sphincter labeled cells in Onuf's nucleus, primarily in L7. Importantly, CTB-labeled LA motor neurons were virtually absent in Onuf's nucleus, where all pudendal motor neurons are located. CTB-labeled processes were observed within Onuf's nucleus, adjacent to FG-labeled pudendal motor neurons, and appeared to derive from dendrites of LA motor neurons that project into Onuf's nucleus. CONCLUSION: The LA muscle has a distinct innervation with very little or no contribution from the pudendal nerve. The intriguing labeling of LA neural elements within a nucleus that innervates the external urethral and anal sphincters (involved in pelvic visceral control) may represent a neuroanatomic substrate for physiologic integration of spinal and supraspinal inputs for the coordination of pelvic floor and visceral activity. PMID- 15902151 TI - Pelvic Organ Support Study (POSST) and bowel symptoms: straining at stool is associated with perineal and anterior vaginal descent in a general gynecologic population. AB - OBJECTIVE: The purpose of this study was to evaluate the association of constipation symptoms and anal incontinence with vaginal wall and pelvic organ descent in a general gynecologic population. STUDY DESIGN: In this multicenter, cross-sectional study, 1004 women attending routine gynecologic healthcare underwent pelvic organ prolapse quantification (POPQ) measurements, and were surveyed regarding anal incontinence, digitation, < 2 bowel movements (BMs)/week, and > 25% frequency of: straining, hard/lumpy stools, and incomplete emptying. Constipation scores reflected the sum of positive responses. Associations between POPQ measurements (Ba, C, Bp, gh+pb), constipation scores, and anal incontinence were evaluated using multivariable regression. RESULTS: Of 119 women with Bp > or = -1.00, 47% reported no constipation symptoms. Hard/lumpy stools (26%), incomplete emptying (24%), and straining (24%) were more prevalent; fewer women reported < 2 BMs/week (15%) or digitation (7%). Constipation scores were weakly correlated with Bp, gh+pb (both r < .1, P < .02). Women reporting > or = 2 symptoms had greater gh+pb measurements than women reporting 0 or 1 symptom (P = .03). Women with anal incontinence had greater gh+pb and gh values than women without anal incontinence (P < .01). POPQ measurements were regressed separately onto (1) total constipation scores, (2) dichotomized scores, and (3) individual symptoms, with BMI, age, number of vaginal deliveries (NVD), weight of largest vaginal delivery (WLVD), race, hysterectomy, study site, and income included as covariates. Total constipation scores and dichotomized scores were nonsignificant in all models. With regard to individual symptoms, straining at stool was significant in the models for Ba and gh+pb, with greater Ba and gh+pb measurements among strainers relative to nonstrainers. CONCLUSION: Most associations between bowel symptoms and vaginal or pelvic organ descent were weak. After controlling for important covariates, straining at stool remained associated with anterior vaginal wall and perineal descent. PMID- 15902152 TI - Estrogen increases collagen I and III mRNA expression in the pelvic support tissues of the rhesus macaque. AB - OBJECTIVE: Our aim was to study the effect of estradiol and raloxifene on collagen synthesis, by measuring the expression collagen I and III mRNA. STUDY DESIGN: Nineteen nulliparous young adult rhesus macaques underwent oophorectomy and were treated for 5 months with estradiol alone, raloxifene, or no hormone. Tissue samples were acquired from the lateral vaginal wall, and included the paravaginal attachment and levator ani muscle. Expression of mRNA for collagen I and III was measured by in situ hybridization. RESULTS: Estradiol increased mRNA for collagen I and III compared with no hormone and raloxifene treatment (ANOVA, P < .05). Collagen mRNA was localized to fibroblasts in the vaginal connective tissue and the connective tissue investments of striated muscle. Collagen mRNA was not expressed in epithelial, smooth, and striated muscle cells. CONCLUSION: Estrogen, but not raloxifene, increases collagen gene transcription and indicates stimulation of collagen synthesis in pelvic floor connective tissues. PMID- 15902153 TI - Low risk of ureteral obstruction with "deep" (dorsal/posterior) uterosacral ligament suture placement for transvaginal apical suspension. AB - OBJECTIVE: Transvaginal uterosacral ligament fixation (USLF), often called "high" USLF, is associated with a 1.0% to 10.9% ureteral obstruction rate. Anatomic relations and pelvic rotation with positioning imply "high" (cephalad) suture placement may bring sutures closer to the ureter. We examined the ureteral obstruction rate with a "deep" (dorsal/posterior) uterosacral ligament suture placement modification of a standard USLF procedure. STUDY DESIGN: At the University of Massachusetts and Tufts, 411 consecutive patients underwent Mayo culdoplasty utilizing > or = 3 uterosacral sutures placed "deep" bilaterally. Intraoperative cystoscopy was performed. RESULTS: One patient (0.24% [.01% 1.35%]) had ureteral obstruction attributable to USLF. Two had obstruction secondary to concomitant procedures. Compared with previous published series, the odds of ureteral injury secondary to USLF was 4.6 times lower (95% CI 2.31-9.24; P < .0001). CONCLUSION: Placement of USLF sutures "deep" (dorsal/posterior) increases the margin of safety for the ureter and, in this study, decreased the ureteral injury rate nearly 5-fold. PMID- 15902154 TI - Incidence and predictors of prolonged urinary retention after TVT with and without concurrent prolapse surgery. AB - OBJECTIVES: The purpose of this study was to describe the time to adequate voiding, incidence of urinary retention, and predictors of voiding efficiency and urinary retention after tension-free vaginal tape (TVT) with and without concurrent prolapse surgery. STUDY DESIGN: Medical records of patients who underwent TVT between August 1999 and July 2003 were reviewed. Urinary retention was defined as the need for urethrolysis, urethral dilation, or postoperative catheterization for >6 weeks. Linear and logistic regression models were used to determine predictors of time to adequate voiding and urinary retention. RESULTS: Two hundred sixty-seven patients were available for analysis; 66% had concurrent prolapse repair, 4% had concurrent laparoscopically assisted vaginal hysterectomy (LAVH), and 30% had an isolated TVT. TVT with and without concurrent prolapse repair or LAVH were statistically similar with respect to median days to voiding (8 vs 5) and the rate of urinary retention (11.2% vs 11.3%). Overall, 4.9% underwent urethrolysis, 1.9% received urethral dilation, and 4.1% required prolonged catheterization. Increasing age, decreasing BMI, and postoperative urinary tract infection were independent predictors of time to adequate voiding. Previous history of incontinence surgery was the only independent predictor of urinary retention (Adjusted odds ratio [AOR] 2.96, 95%CI [1.17-7.06]). CONCLUSION: Concurrent prolapse surgery does not appear to significantly alter postoperative voiding efficiency or increase the risk of prolonged urinary retention compared with TVT alone. PMID- 15902155 TI - Anatomic relationship between the vaginal apex and the bony architecture of the pelvis: a magnetic resonance imaging evaluation. AB - OBJECTIVE: This study was undertaken to define anatomic relationships between the vaginal apex and the ischial spines and sacrum for nulliparous women with normal support. STUDY DESIGN: We retrospectively evaluated the magnetic resonance images of 11 consecutive women who underwent pelvic imaging at Johns Hopkins. Coordinates were recorded for the posterior fornix, sacrum, ischial spines, and cervical vaginal junctions. We calculated vector distances with means, SDs, and 95% CIs. Intraclass correlation coefficients tested interobserver reliability and the Wilcoxon signed rank test compared right- and left-sided measurements. RESULTS: Mean age was 30.4 +/- 9.1 years. The cervical vaginal junction was 1.6 +/- 0.5 cm superior, 1.1 +/- 0.5 cm anterior, and 4.7 +/- 0.4 cm medial to the ipsilateral ischial spine. The posterior fornix was 1.0 +/- 1.0 cm anterior and 5.3 +/- 0.8 cm inferior to the second sacral vertebra. There was excellent interobserver reliability (interclass correlation coefficients = 0.997, P < .001) and no detectable difference between sides. CONCLUSION: Consistent relationships exist between the vaginal apex and ischial spines and sacrum, which may be useful in reconstructive pelvic surgery. PMID- 15902156 TI - Capsaicin for the treatment of vulvar vestibulitis. AB - OBJECTIVE: The purpose of this study was to evaluate the use of local capsaicin cream as an effective treatment for patients with documented vulvar vestibulitis syndrome. STUDY DESIGN: A retrospective chart review was performed for patients who received a diagnosis of vulvar vestibulitis syndrome that was treated with capsaicin. Patients performed local application of capsaicin 0.025% cream for 20 minutes daily for 12 weeks. A comparison was made between the pre- and posttreatment Kaufman touch test to evaluate discomfort. The Marinoff dyspareunia scale was also used to assess pre- and posttreatment. RESULTS: The sum of the Kaufman touch test scores before the treatment (13.2 +/- 4.9) compared with the scores after treatment (4.8 +/- 3.8) was statistically improved (P < .001). A significant improvement was also observed at each individual site (P < .001). The Marinoff dyspareunia scale also showed a significant improvement (P < .001). CONCLUSION: Vulvar vestibulitis syndrome that is treated with capsaicin significantly decreases discomfort and allows for more frequent sexual relations. PMID- 15902157 TI - The usefulness of urinary cytology testing in the evaluation of irritative voiding symptoms. AB - OBJECTIVE: The purpose of this study was to assess the clinical usefulness of urinary cytology testing for the evaluation of urothelial cancer in women with irritative voiding symptoms who were examined at a urogynecology service. STUDY DESIGN: Urinary cytology studies results that were obtained from January 1, 2000, to December 31, 2002, were cross-matched with the Rhode Island Department of Health Cancer Registry to identify those women who were diagnosed with urinary tract malignancies. The prevalence of urothelial cancer was determined, and the sensitivity, specificity, and positive and negative predictive values of urinary cytologic testing were calculated for 2 common classification strategies: (1) consideration of atypical cytologic test results to be normal and (2) consideration of atypical cytologic test results to be abnormal. RESULTS: Among 1516 cross-matched cytologic test results from 1324 patients, 5 urothelial cancers were identified. Two of the 5 malignancies were associated with positive cytology results. The prevalence of urothelial cancer was 0.38% (95% CI, 0.1%, 0.9%). When atypical cytology studies were classified as normal, the sensitivity of urinary cytology was 40% (95% CI, 7.2%, 83.0%); the specificity was 99.9% (95% CI, 99.5%, 100%); the positive predictive value was 66.7% (95% CI, 12.5%, 98.2%), and negative predictive value was 99.8% (95% CI, 99.2%, 100%). In contrast, when atypical cytology results were classified as abnormal, the sensitivity and negative predictive value remained the same, but the specificity declined to 93.6% (95% CI, 92.1%, 94.8%), and the positive predictive value decreased to 2.3% (95% CI, 0.4%, 8.8%). CONCLUSION: The low prevalence of urothelial cancers and low sensitivity of urinary cytology studies severely limit the usefulness of this test in the evaluation of women with irritative voiding symptoms. PMID- 15902158 TI - Results of urine cytology testing and cystoscopy in women with irritative voiding symptoms. AB - OBJECTIVE: The purpose of this study was to assess rates of urinary cytologic abnormalities and cystoscopic outcomes in women with irritative voiding symptoms who were examined at a urogynecology clinic. STUDY DESIGN: All urinary cytology studies results that were sent between January 1, 2000, and July 31, 2003, for the evaluation of irritative voiding symptoms were reviewed. Data were then extracted from the charts of a subset of these patients to evaluate cystoscopic outcomes. Demographics, risk factors for urothelial cancer, laboratory results, and radiology imaging results were then analyzed and compared between patients with and without abnormal cytology and cystoscopic results. RESULTS: Of the 1783 total urinary cytology that were reviewed, 1661 test results were read as normal (93.2%); 112 test results (6.3%) were read as atypical, and 3 test results (0.2%) were read as unsatisfactory. Seven cytologic test results were categorized as suspicious or malignant, which accounts for only 0.4% of all cytologic test results that were sent. Of the 564 consecutive women whose cases were chosen for subanalysis, cytology was normal in 91.5% and atypical in 8.5% of cases. No cytology were suspicious or malignant. Cystoscopic findings were normal in 548 patients (97.2%). Only 1 patient (0.2%) received a diagnosis of transitional cell carcinoma. CONCLUSION: Urinary cytology and cystoscopy are low yield tests and should not be used routinely in the initial evaluation of women with irritative voiding symptoms. PMID- 15902159 TI - Suburethral sling treatment of occult stress incontinence and intrinsic sphincter deficiency in women with severe vaginal prolapse of the anterior vs posterior/apical compartment. AB - OBJECTIVE: The purpose of this study was to compare the efficacy of a Mersilene mesh suburethral sling for occult stress urinary incontinence (SUI) and intrinsic sphincter deficiency (ISD) in women with severe vaginal prolapse of the anterior compartment to the posterior/apical compartment. STUDY DESIGN: This was a retrospective study that compared women with stage or grade III/IV prolapse of the anterior compartment (group 1) with the posterior/apical compartment (group 2); both groups demonstrated occult SUI (leakage only with prolapse reduced) and ISD on urodynamics, and underwent concurrent pelvic reconstructive surgery. The sling was defined as efficacious if SUI was prevented in 85% of women and if obstructive symptoms (de novo or worsening urge incontinence, or urinary retention greater than 2 weeks) occurred in less than 10% of women. RESULTS: There were 39 women in group 1 and 25 women in group 2. There were no differences between women in group 1 or group 2 in preoperative demographics (except parity) or urodynamic findings. SUI cure rates were lower for group 1 than group 2, but this difference was not significant (87% vs 100%, P = .15). Rates of de novo or worsening urge incontinence (8% vs 4%, P = 1.00) and urinary retention (none occurred) were similar between groups. CONCLUSION: In women with severe vaginal prolapse, slings effectively treat occult SUI and ISD, whether associated with anterior or posterior/apical prolapse. PMID- 15902160 TI - Do alterations in vaginal dimensions after reconstructive pelvic surgeries affect the risk for dyspareunia? AB - OBJECTIVE: The purpose of this study was to determine whether changes in vaginal dimensions after transvaginal reconstructive pelvic surgeries affect the risk for postoperative dyspareunia. STUDY DESIGN: Charts of all sexually active patients who underwent transvaginal reconstructive pelvic surgeries in our institution between July 1998 and June 2002 with 1 year of follow-up evaluations were reviewed. Data were analyzed with the Student t, chi2, Pearson's correlation tests and a logistic regression model. RESULTS: Two hundred twenty-eight women aged 44 to 83 years were included. Dyspareunia increased after operation (16% vs 7%; P = .001); total vaginal length (7.6 vs 8.8 cm; P = .001) and genital hiatus (2.7 vs 3.5 cm; P = .001) dimensions significantly decreased after operation, with no predilection for any specific procedure. No correlation was found between these changes in vaginal dimensions and the risk for dyspareunia. CONCLUSION: The prevalence of dyspareunia increases after transvaginal reconstructive pelvic surgeries. Despite a postoperative decrease in vaginal dimensions, a causal relationship between dyspareunia and changes in vaginal dimensions may not exist. PMID- 15902161 TI - A randomized, double-blinded, sham-controlled trial of postpartum extracorporeal magnetic innervation to restore pelvic muscle strength in primiparous patients. AB - OBJECTIVE: The purpose of this study was to determine the effects of extracorporeal magnetic innervation (ExMI) on pelvic muscle strength of primiparous patients. STUDY DESIGN: Primigravid patients were randomized to receive either active or sham ExMI postpartum treatments for 8 weeks. The main outcome measure was pelvic muscle strength measured by perineometry at baseline (midtrimester), 6 weeks (before treatments), 14 weeks, 6 months, and 12 months postpartum. Mixed randomized-repeated measures ANOVA was used to analyze the mean perineometry values between the 2 groups and across all 5 time periods. RESULTS: Fifty-one patients enrolled, and 18 were lost to attrition. There were no differences in demographics or delivery characteristics between the active and sham groups. There was an overall time effect, F(3,85) = 3.1, P = .049, but no group, F(1,31) = 0.007, P = .94, or (group)(time) interaction, F(3,85) = 1.8, P = .15. CONCLUSION: We found no differences in pelvic muscle strength between patients receiving active or sham ExMI treatments in the early postpartum period. PMID- 15902162 TI - A comparison of perineometer to brink score for assessment of pelvic floor muscle strength. AB - OBJECTIVE: The Brink scale is a commonly used digital assessment of pelvic floor muscle strength. The Peritron perineometer, a compressible vaginal insert that records pressure in centimeters of water, offers an objective method for this evaluation. This study evaluates the inter- and intrarater reliability of perineometry measurements and correlates those values with Brink scores. STUDY DESIGN: Subjects were prospectively enrolled and underwent pelvic floor muscle strength assessment by 2 examiners each using a perineometer and the Brink scale. Perineometer measurements of maximum pressure, average pressure, and total duration were recorded for 3 consecutive pelvic floor muscle contractions (Kegels). The Brink assessment was performed by placing 2 fingers vaginally during a single Kegel contraction. Brink scores consisted of 3 separate 4-point rating scales for pressure, vertical finger displacement, and duration. The order of the examiners and the 2 assessment methods were randomized, and each examiner was blinded to the results of the other. Pearson and Spearman correlation coefficients were used for analysis as appropriate. Repeated-measures analysis of variance was used to assess intrarater reliability between repeated perineometer measurements. RESULTS: One hundred women were consecutively enrolled and completed the study. Interrater reliability for the perineometer maximum squeeze pressure (r = 0.88) and baseline resting pressure (r = 0.78) was high. Maximum squeeze pressure correlation was unaffected by the presence or absence of estrogen (r = 0.89 versus r = 0.85), nulliparity versus parity (0.85 versus 0.88), or genital hiatus 4 or greater or less than 4 (r = 0.96 versus r = 0.86). Total Brink score and each individual submeasurement showed good correlations (total: r = 0.68; pressure: r = 0.68; displacement: r = 0.58; duration: r = 0.44). The correlation between maximum squeeze pressure and total Brink score during the first and second exams was good (r = 0.68 versus r = 0.71). For intrarater reliability, there were no significant differences among the 3 maximum squeeze pressures recorded during the first exam (P = .11), but for the second exam, the first squeeze was significantly stronger than the successive 2 (P = .009) attempts. CONCLUSION: Perineometer measurements of pelvic floor muscle contractions show very good inter- and intrarater reliability. The Brink total and pressure scores had a slightly lower interrater reliability. Variables such as estrogen status, parity, and genital hiatus did not appear to affect correlation. There was good correlation between the maximum perineometer pressure and the total Brink score, suggesting that these 2 methods of assessment have similar levels of reproducibility. Additionally, the perineometer demonstrated good short-term test-retest reliability. PMID- 15902163 TI - Levator contraction strength and genital hiatus as risk factors for recurrent pelvic organ prolapse. AB - OBJECTIVE: To correlate levator ani contraction strength and genital hiatus measurements with surgical failure in prolapse. STUDY DESIGN: This retrospective study involved chart review for documentation of levator contraction strength, genital hiatus measurement, and recurrent pelvic floor disorders in women who underwent surgery for prolapse. RESULTS: The recurrent prolapse rate was 34.6%. Median follow-up interval was 5 months. Diminished levator strength was associated with recurrent prolapse (35.8% versus 0%; P = .017). A genital hiatus 5 cm or greater was associated with recurrent prolapse (44.2% vs 27.8%; P = .034). Inability to contract the levator ani was associated with urinary incontinence (35.1% vs 18.8%; P = .023). Increasing levator contraction strength was associated with a decreased reoperation rate for pelvic floor disorders, whereas genital hiatus correlated best with recurrent prolapse. CONCLUSION: Diminished levator ani contraction strength and a widened genital hiatus correlate with an increase in surgical failures in the early postoperative period. These tools are useful for counseling a patient concerning surgery for prolapse. PMID- 15902164 TI - The incidence of urinary tract injury during hysterectomy: a prospective analysis based on universal cystoscopy. AB - OBJECTIVE: To evaluate the incidence of urinary tract injury due to hysterectomy for benign disease. STUDY DESIGN: Patients were enrolled prospectively from 3 sites. All patients undergoing abdominal, vaginal, or laparoscopic hysterectomy for benign disease underwent diagnostic cystourethroscopy. RESULTS: Four hundred seventy-one patients participated. Ninety-six percent (24/25) of urinary tract injuries were detected intraoperatively. There were 8 cases of ureteral injury (1.7%) and 17 cases of bladder injury (3.6%). Ureteral injury was associated with concurrent prolapse surgery (7.3% vs 1.2%; P = .025). Bladder injury was associated with concurrent anti-incontinence procedures (12.5% vs 3.1%; P = .049). Abdominal hysterectomy was associated with a higher incidence of ureteral injury (2.2% vs 1.2%) but this was not significant. Only 12.5% of ureteral injuries and 35.3% of bladder injuries were detected before cystoscopy. CONCLUSION: The incidence of urinary tract injury during hysterectomy is 4.8%. Surgery for prolapse or incontinence increases the risk. Routine use of cystoscopy during hysterectomy should be considered. PMID- 15902165 TI - Elastin metabolism in pelvic tissues: is it modulated by reproductive hormones? AB - OBJECTIVE: The purpose of this study was to investigate the effect of relaxin on extracellular matrix protein expression in pelvic fibroblasts that were cultured from women with stress urinary incontinence compared with asymptomatic control subjects. STUDY DESIGN: Periurethral vaginal wall fibroblasts from premenopausal women with stress urinary incontinence and continent women (in both the proliferative and secretory phase of the menstrual cycle) were stimulated with increasing concentrations of relaxin (0-500 ng/mL). The supernatant was sampled for matrix metalloproteinase-2 and -9 by zymography. Tissue inhibitors of metalloproteinase-1 and -2 and alpha-1 antitrypsin were evaluated with Western blot. Total elastase activity was measured by generation of free amino groups from succinylated elastin. Increasing concentrations of alpha-1 antitrypsin were added to cell lysate to evaluate total elastase activity inhibition. RESULTS: Proliferative-phase stress urinary incontinence fibroblasts demonstrated an increase in matrix metalloproteinase-2 and no change in matrix metalloproteinase 9 and tissue inhibitors of metalloproteinase-1 and -2 expressions with increasing relaxin concentrations. Cells from control subjects showed increased expression of matrix metalloproteinase-2 and -9, but no change in tissue inhibitors of metalloproteinases. Secretory-phase stress urinary incontinence fibroblasts showed no response in matrix metalloproteinase or tissue inhibitors of metalloproteinase expressions with relaxin stimulation. Secretory-phase control fibroblasts reacted by increasing matrix metalloproteinase-2 and -9 and tissue inhibitors of metalloproteinase-2. With respect to total elastase activity and alpha-1 antitrypsin expression, increasing doses of relaxin appear to increase elastolytic activity in stress urinary incontinence cells by decreasing the expression of alpha-1 antitrypsin in proliferative phase cells or increasing the total elastase activity in secretory phase cells. Fibroblast total elastase activity was inhibited by increasing concentrations of alpha-1 antitrypsin. CONCLUSION: Elastase activity appears to be increased in relaxin-stimulated stress urinary incontinence fibroblasts by either decreased inhibitor (alpha-1 antitrypsin) production or increased elastase activity. PMID- 15902166 TI - The safety and efficacy of laparoscopic surgical staging of apparent stage I ovarian and fallopian tube cancers. AB - OBJECTIVE: To compare the safety and efficacy of laparoscopic staging of ovarian or fallopian tube cancers to staging via laparotomy for epithelial ovarian carcinoma. STUDY DESIGN: We performed a case-control study of all patients with apparent stage I adnexal cancers who had laparoscopic staging from October 2000 to March 2003. The control group consisted of all patients with apparent stage I epithelial ovarian carcinoma who had staging via laparotomy during the same time period. RESULTS: Staging was laparoscopic in 20 patients and via laparotomy in 30. There were no differences in mean age and body mass index. There were also no differences in omental specimen size and number of lymph nodes removed. Estimated blood loss and hospital stay were lower for laparoscopy, but operating time was longer. There were no conversions to laparotomy or complications in the laparoscopic group, compared with 3 minor complications in the laparotomy group. CONCLUSION: In this preliminary analysis, it appears that patients with apparent stage I ovarian or fallopian tube cancer can safely and adequately undergo laparoscopic surgical staging. PMID- 15902167 TI - Decreased anal sphincter lacerations associated with restrictive episiotomy use. AB - OBJECTIVE: To determine whether restrictive episiotomy use was associated with decreases in anal sphincter lacerations and the risk of anal sphincter laceration attributable to episiotomy. STUDY DESIGN: This was a retrospective database study. Rates of episiotomy, anal sphincter laceration (third- or fourth-degree tear), and other confounding variables were compared among vaginal deliveries before (1999) and after (2002) restrictive episiotomy use was implemented at our institution. Logistic regression was used to estimate the odds ratio of anal sphincter laceration that was due to episiotomy and other variables. RESULTS: The episiotomy rate decreased 56% (37% to 17%, P < .001) between 1999 and 2002, whereas the anal sphincter laceration rate decreased 44% (9.7% to 5.4%, P < .001). There were no changes in age, race, nulliparity, prolonged second stage of labor, operative vaginal deliveries, birth weight, or macrosomia, although oxytocin use and epidural use decreased slightly (37% to 31%, P < .001, and 80% to 76%, P = .02, respectively). The adjusted odds ratio of anal sphincter laceration attributable to episiotomy decreased 55%, from 6.5 (95% CI: 3.8, 11.1) to 2.9 (95% CI: 1.7, 5.0), between 1999 and 2002. Conversely, the adjusted odds ratios of anal sphincter laceration attributable to the other independent risk factors all increased or remained the same: operative vaginal delivery, which increased from 4.4 (95% CI: 2.7, 6.9) to 6.3 (95% CI: 3.6 11.1); nulliparity, from 2.9 (95% CI: 1.8, 4.8) to 2.9 (95% CI: 1.4, 5.9); macrosomia, from 1.9 (95% CI: 1.1, 3.4) to 2.6 (95% CI: 1.3, 5.4); and prolonged second stage, from 2.0 (95% CI: 1.3, 3.0) to 2.1 (95% CI: 1.2, 3.7). CONCLUSION: With restrictive episiotomy use, the episiotomy rate, anal sphincter laceration rate, and risk of anal sphincter laceration attributable to episiotomy were all reduced by approximately 50%. PMID- 15902168 TI - Suture erosion and wound dehiscence with permanent versus absorbable suture in reconstructive posterior vaginal surgery. AB - OBJECTIVE: This study was undertaken to determine the incidence of wound disruption after reconstructive posterior vaginal surgery with braided permanent versus absorbable suture. STUDY DESIGN: A retrospective cohort study of women undergoing posterior vaginal surgery. Outcomes included suture erosion, wound dehiscence, and additional surgical procedures. RESULTS: Ninety-nine procedures were performed with permanent sutures, followed by 111 with absorbable sutures. There were no differences in demographics or comorbidities between patient groups. Suture erosion/wound dehiscence occurred in 31.3% of the permanent suture group versus 9% of the absorbable suture group (P = .003, odds ratio [OR] = 7.5, 95% CI 2-28). The need for additional surgical intervention was 16.1% among permanent suture group versus no patients with absorbable suture. Performing a concomitant anal sphincteroplasty with permanent sutures significantly increased the incidence of suture erosion (P = .003, OR = 4.7, 95%CI 1.7-13.3). CONCLUSION: Permanent sutures increase the incidence in wound disruption and the need for additional surgical intervention in posterior colporrhaphy and anal sphincteroplasty. PMID- 15902169 TI - Incidence of perioperative complications of urogynecologic surgery in elderly women. AB - OBJECTIVE: The purpose of this study was to determine the incidence of and risk factors for perioperative complications in elderly women who undergo urogynecologic surgery. STUDY DESIGN: A retrospective chart review of patients > or = 75 years old who underwent urogynecologic surgery between January 1999 and December 2003 was performed. Demographics, comorbidities, and significant perioperative complications were recorded. The Charlson Comorbidity Index and American Society of Anesthesiologists classification were calculated to summarize the patients' overall perioperative risk. Logistic regression was used to identify independent risk factors for perioperative complications. RESULTS: Two hundred sixty-seven patients who were > or = 75 years old met the inclusion criteria; 25.8% of the patients had a significant perioperative complication. The most common perioperative complication was blood transfusion or significant blood loss, pulmonary edema, and postoperative congestive heart failure. Independent risk factors that were predictive of a patient having a perioperative complication were the length of surgery, coronary artery disease, and peripheral vascular disease. The Charlson Comorbidity Index and American Society of Anesthesiologists classification did not predict perioperative complications in this population. CONCLUSION: Pre-existing cardiovascular disease increases the risk of a significant perioperative complication in elderly women who undergo urogynecologic surgery. However, the overall perioperative morbidity rate in elderly women who undergo urogynecologic surgery is low. PMID- 15902170 TI - Anal incontinence in women presenting for gynecologic care: prevalence, risk factors, and impact upon quality of life. AB - OBJECTIVE: The purpose of this study was to estimate the prevalence and impact upon quality of life of anal incontinence (AI) in women aged 18 to 65. STUDY DESIGN: Consecutive women presenting for general gynecologic care were given a bowel function questionnaire. Women with AI were prompted to complete the Fecal Incontinence Severity Index (FISI) and Fecal Incontinence Quality of Life Scale (FIQL). RESULTS: The cohort was composed of 457 women with a mean age of 39.9 +/- 11 years. AI prevalence was 28.4% (95% CI 24.4-32.8). After logistic regression, IBS (OR 3.22, 1.75-5.93), constipation (OR 2.11, 1.22-3.63), age (OR 1.05, 1.03 1.07), and BMI (OR 1.04, 1.01-1.08) remained significant risk factors. The mean FISI score was 20.4 +/- 12.4. Women with only flatal incontinence scored higher, and women with liquid loss scored lower on all 4 scales of the FIQL. CONCLUSION: AI is prevalent in women seeking benign gynecologic care, and liquid stool incontinence has the greatest impact upon quality of life. PMID- 15902171 TI - Histopathologic changes of porcine dermis xenografts for transvaginal suburethral slings. AB - OBJECTIVE: The purpose of this study was to examine the histopathologic changes of HMDI (Hexamethylene di-isocyanate) cross-linked porcine dermis grafts used for suburethral sling surgery. STUDY DESIGN: Twelve patients underwent reoperation with graft removal for urinary retention or recurrent stress urinary incontinence after transvaginal sling surgery. Tissue specimens were available for pathologic evaluation in 7 patients. Graft specimens underwent histologic preparation including hematoxylin and eosin staining. A single pathologist reviewed the slides blinded to clinical outcomes. RESULTS: Histopathologic analyses revealed only limited collagen remodeling, and evidence of a foreign body type reaction was present in some specimens. In cases of recurrent stress incontinence, implants appeared to be completely replaced by dense fibroconnective tissue and moderate neovascularization without evidence of inflammation or graft remnants. CONCLUSION: HMDI cross-linked porcine dermal collagen implants result in variable tissue reactions that may have unpredictable clinical outcomes in different patients, raising questions about the overall tolerability and efficacy of these grafts in pelvic reconstructive surgery. PMID- 15902172 TI - A prospective randomized trial using solvent dehydrated fascia lata for the prevention of recurrent anterior vaginal wall prolapse. AB - OBJECTIVE: This study was undertaken to compare outcomes after anterior colporrhaphy with and without a solvent dehydrated cadaveric fascia lata graft. STUDY DESIGN: A total of 162 women were enrolled in a prospective, randomized trial that evaluated the impact of a solvent dehydrated cadaveric fascia lata patch on recurrent anterior vaginal prolapse. Subjects were randomly assigned to standard colporrhaphy with or without a patch. Before and after surgery, subjects were evaluated by both the Baden-Walker and pelvic organ prolapse quantification systems. "Failure" was defined as stage II anterior wall prolapse or worse. RESULTS: Of 154 women randomly assigned (76 patch: 78 no patch), all underwent surgery and 153 (99%) returned for follow-up. Sixteen women (21%) in the patch group and 23 (29%) in the control group experienced recurrent anterior vaginal wall prolapse (P = .229). Only 26% of all recurrences were symptomatic. Concomitant transvaginal Cooper's ligament sling procedures were associated with a dramatic decrease in recurrent prolapse (odds ratio [OR] 0.105 , P < .0001). CONCLUSION: Solvent dehydrated fascia lata as a barrier does not decrease recurrent prolapse after anterior colporrhaphy. Transvaginal bladder neck slings were associated with a significant reduction in the risk of recurrent anterior wall prolapse. PMID- 15902173 TI - Obstetric antecedents for postpartum pelvic floor dysfunction. AB - OBJECTIVE: The purpose of this study was to evaluate prospectively the association between selected obstetric antecedents and symptoms of pelvic floor dysfunction in primiparous women up to 7 months after childbirth. STUDY DESIGN: All nulliparous women who were delivered between June 1, 2000, and August 31, 2002, were eligible for a postpartum interview regarding symptoms of persistent pelvic floor dysfunction. Responses from all women who completed a survey at or before their 6-month contraceptive follow-up visit were analyzed. Obstetric antecedents to stress, urge, and anal incontinence were identified, and attributable risks for each factor were calculated. RESULTS: During the study period, 3887 of 10,643 primiparous women (37%) returned within 219 days of delivery. Symptoms of stress and urge urinary incontinence, were significantly reduced (P < .01) in women who underwent a cesarean delivery. Symptoms of urge urinary incontinence doubled in women who underwent a forceps delivery (P = .04). Symptoms of anal incontinence were increased in women who were delivered of an infant who weighed >4000 g (P = .006) and more than doubled in those women who received oxytocin and had an episiotomy performed (P = .01). CONCLUSION: The likelihood of symptoms of pelvic floor dysfunction up to 7 months after delivery was greater in women who received oxytocin, who underwent a forceps delivery, who were delivered of an infant who weighed >4000 g, or who had an episiotomy performed. Women who underwent a cesarean delivery had fewer symptoms of urge and stress urinary incontinence. PMID- 15902174 TI - Pudendal neuralgia, a severe pain syndrome. AB - OBJECTIVE: To describe the clinical and electrodiagnostic findings, therapies, and outcomes of patients with pudendal neuralgia. STUDY DESIGN: A retrospective, descriptive study of 64 patients from March 19 to December 22, 2003. RESULTS: Clinical findings included pain along nerve distribution (64, 100%), pain aggravated by sitting (62, 97%), pain relieved by standing or lying (57, 89%), and misdiagnosis (53, 83%). Neurophysiologic findings were normal (23, 35%), demyelination (17, 26%), axonal loss (5, 7.5%), and demyelination with axonal loss (21, 32%). Therapies were conservative (64, 100%), nerve injection (38, 59%), neuromodulation (2, 3%), and decompression surgery (10, 15%). Slight or moderate pain improvement with therapies included conservative (64, 100%), nerve injection (12, 31%), neuromodulation (2, 100%), and decompression (6, 60%). CONCLUSION: Pudendal neuralgia is poorly recognized and poorly treated. Improvement is gained with conservative therapy. Injections and decompression benefit one half and one third of patients, respectively. Neuromodulation needs further evaluation. PMID- 15902175 TI - Pudendal nerve stretch during vaginal birth: a 3D computer simulation. AB - OBJECTIVE: The purpose of this study was to determine the increase in pudendal nerve branch lengths using a 3D computer model of vaginal delivery. STUDY DESIGN: The main inferior rectal and perineal branches of the pudendal nerve were dissected in 12 hemi-pelves from 6 adult female cadavers. Their 3D courses were digitized in the 4 specimens with the most characteristic nerve branching pattern, and the data were imported into a published 3D computer model of the pelvic floor. Each nerve branch was then represented by a stretchable cord with a fixation point at the ischial spine. The length change in each branch was then quantified as the fetal head descended through the pelvic floor. The maximum nerve strains ([final length minus original length/original length] x 100) were calculated for 5 degrees of perineal descent: reference descent from the literature, 1.25 cm and 2.5 cm caudal and cephalad. The effect of alternative fixation points on resultant nerve strain was also studied. RESULTS: The inferior rectal branch exhibited the maximum strain, 35%, and this strain varied by 15% from the scenario with the least perineal descent to that with the most perineal descent. The strain in the perineal nerve branch innervating the anal sphincter reached 33%, while the branches innervating the posterior labia and urethral sphincter reached values of 15% and 13%, respectively. The more proximal the nerve fixation point, the greater the nerve strain. CONCLUSION: During the second stage: (1) nerves innervating the anal sphincter are stretched beyond the 15% strain threshold known to cause permanent damage in appendicular peripheral nerve, and (2) the degree of perineal descent is shown to influence pudendal nerve strain. PMID- 15902176 TI - Comparison of long-term outcomes of autologous fascia lata slings with Suspend Tutoplast fascia lata allograft slings for stress incontinence. AB - OBJECTIVE: This study was undertaken to compare the objective and subjective long term surgical outcomes in patients receiving Tutoplast fascia lata allograft slings with those receiving autograft slings for the treatment of stress urinary incontinence (SUI). STUDY DESIGN: We reviewed all patients (n = 71) undergoing suburethral sling with either autologous fascia lata (n = 39) or Tutoplast fascia lata (n = 32) for urodynamic stress incontinence (USI) from October 1, 1998, to August 1, 2001. RESULTS: Of the original 71 patients, 47 were evaluated by objective and/or subjective means at a minimum of 2 years after surgery. Subjective quality of life measures, subjective continence, maximum urethral closure pressure, and bladder neck mobility were not different between the 2 groups. USI was demonstrated in 41.7% of allograft patients compared with no autograft patients (P = .007). CONCLUSION: Although patient reported cure of SUI is high for both sling types, USI recurs at a significantly higher rate in Tutoplast slings compared with autologous slings. PMID- 15902177 TI - Nocturnal polyuria in women with overactive bladder symptoms and nocturia. AB - OBJECTIVE: This study was undertaken to determine the prevalence of nocturnal polyuria in women complaining of nocturia and overactive bladder (OAB) symptoms and to identify clinical and health characteristics associated with nocturnal polyuria. STUDY DESIGN: Women presenting to a urogynecology clinic with complaints of nocturia and OAB symptoms were asked to participate. They completed a health characteristic summary, 3-day voiding diary, Nordic sleep questionnaire, urinary distress inventory (UDI), and a nocturia distress visual log (NDVL). The 24-hour urine production, nighttime urine volume, and maximum bladder capacity were calculated from the bladder diary. Nocturnal polyuria was defined as production of greater than 33% of the 24-hour urine volume during an 8-hour sleep period. A histogram was performed to analyze at what age the prevalence of nocturnal polyuria increased. Women were then divided by presence or absence of nocturnal polyuria and compared by the health and clinical characteristics. RESULTS: Fifty-five women met the qualifications and participated in the study. Average age of the cohort was 65.8 +/- 13.5 years. The risk of nocturnal polyuria increased with age 65 years or older (prevalence for age 65-74 = 0.86 [0.62 1.00]) and with white race. On the basis of the mean population values for UDI, NDVL, sleepiness scores, and insomnia scores, all women were bothered by their nocturia. Median number of nighttime voids, 24-hour urine production, maximum bladder capacity, nocturnal index, UDI, NDVL, sleepiness scores, and insomnia scores did not differ, based on presence or absence of nocturnal polyuria. CONCLUSION: Among women complaining of nocturia and overactive bladder symptoms, age 65 years or older and white race appear to be risk factors for nocturnal polyuria. PMID- 15902178 TI - Pelvic organ prolapse, constipation, and dietary fiber intake in women: a case control study. AB - OBJECTIVE: This study was undertaken to determine whether there is an association among pelvic organ prolapse, constipation, and dietary fiber intake. STUDY DESIGN: Sixty consecutive women with prolapse were compared with 30 control women without prolapse. All women completed 2 validated questionnaires to assess constipation and dietary fiber intake. Multivariate analysis was performed. RESULTS: The risk for constipation was greater in women with prolapse than controls (odds ratio 4.03, 95% CI 1.5-11.4). Median insoluble fiber intake was significantly lower in women with prolapse (2.4 g) than controls (5.8 g, P < .01). The increased risk for constipation was reduced but remained significant after controlling for age and insoluble dietary fiber intake (odds ratio 2.9, 95% CI 1.1-13.5). CONCLUSION: Women with pelvic organ prolapse are at a higher risk for constipation than controls. This increased risk for constipation is partially explained by lower intake of dietary insoluble fiber by women with prolapse than controls. PMID- 15902179 TI - A randomized trial of the effects of coached vs uncoached maternal pushing during the second stage of labor on postpartum pelvic floor structure and function. AB - OBJECTIVE: The purpose of this study was to determine if refraining from coached pushing during the second stage of labor affects postpartum urogynecologic measures of pelvic floor structure and function. STUDY DESIGN: Nulliparous women at term were randomized to coached (n = 67) vs uncoached (n = 61) pushing. At 3 months' postpartum women underwent urodynamic testing, pelvic organ prolapse examination (POPQ), and pelvic floor neuromuscular assessment. RESULTS: Urodynamic testing revealed decreased bladder capacity (427 mL vs 482 mL, P = .051) and decreased first urge to void (160 mL vs 202 mL, P = .025) in the coached group. Detrusor overactivity increased 2-fold in the coached group (16% vs 8%), although this difference was not statistically significant (P = .17). Urodynamic stress incontinence was diagnosed in the coached group in 11/67 (16%) vs 7/61 (12%) in the uncoached group (P = .42). CONCLUSION: Coached pushing in the second stage of labor significantly affected urodynamic indices, and was associated with a trend towards increased detrusor overactivity. PMID- 15902180 TI - Primary repair of obstetric anal sphincter laceration: a randomized trial of two surgical techniques. AB - OBJECTIVE: This study was undertaken to compare surgical techniques for the primary repair of obstetric anal sphincter lacerations. STUDY DESIGN: Patients with complete third- or fourth-degree lacerations were recruited and randomly assigned to either an end-to-end or overlapping repair. Data collection included demographic data, obstetric history, and intrapartum events. Postpartum, women completed incontinence questionnaires and underwent physical and ultrasound examinations. To detect a 36% difference between groups with an alpha = .05 and beta = .20, 30 patients were required. Data were analyzed with Student t test and chi2 analysis. RESULTS: Forty-one women were randomly assigned; 23 to an end-to end and 18 to an overlapping repair. Twenty-seven percent of women underwent episiotomy and 61% operative vaginal delivery. Follow-up was limited to 26 of 41 patients. On physical examination, 3 patients had a separated anal sphincter. On ultrasound, overall 85% of patients had intact sphincters, with no difference between groups (all P > .05). Forty-two percent of women complained of anorectal symptoms with no differences between groups (all P > .28). CONCLUSION: We found no difference in anal incontinence symptoms, physical examination, or translabial ultrasonography findings between the 2 groups. Incontinence symptoms were common in both groups. PMID- 15902181 TI - The impact of occiput posterior fetal head position on the risk of anal sphincter injury in forceps-assisted vaginal deliveries. AB - OBJECTIVE: A forceps-assisted vaginal delivery is a well-recognized risk factor for anal sphincter injury. Some studies have shown that occiput posterior (OP) fetal head position is also associated with an increased risk for third- or fourth-degree lacerations. The objective of this study was to assess whether OP position confers an incrementally increased risk for anal sphincter injury above that present with forceps deliveries. STUDY DESIGN: This was a retrospective cohort study of 588 singleton, cephalic, forceps-assisted vaginal deliveries performed at our institution between January 1996 and October 2003. Maternal demographics, labor and delivery characteristics, and neonatal factors were examined. Statistical analysis consisted of univariate statistics, Student t test, chi2, and logistic regression. RESULTS: The prevalence of occiput anterior (OA) and OP positions was 88.4% and 11.6%, respectively. The groups were similar in age, marital status, body mass index, use of epidural, frequency of inductions, episiotomies, and shoulder dystocias. The OA group had a higher frequency of rotational forceps (16.2% vs 5.9%, P = .03), greater birth weights (3304 +/- 526 g vs 3092 +/- 777 g, P = .004), and a larger percentage of white women (48.8% vs 34.3%, P = .04). Overall, 35% of forceps deliveries resulted in a third- or fourth-degree laceration. Anal sphincter injury occurred significantly more often in the OP group compared with the OA group (51.5% vs 32.9%, P = .003), giving an odds ratio of 2.2 (CI: 1.3-3.6). In a logistic regression model that controlled for occiput posterior position, maternal body mass index, race, length of second stage, episiotomy, birth weight, and rotational forceps, OP head position was 3.1 (CI: 1.6-6.2) times more likely to be associated with anal sphincter injury than OA head position. CONCLUSION: Forceps-assisted vaginal deliveries have been associated with a greater risk for anal sphincter injury. Within this population of forceps deliveries, an OP position further increases the risk of third- or fourth-degree lacerations when compared with an OA position. PMID- 15902182 TI - The distance between the perceived and the actual arcus tendineus fascia pelvis during vaginal paravaginal repair. AB - OBJECTIVE: This study was undertaken to determine whether the arcus tendineus fascia pelvis (ATFP) can be accurately identified from the paravaginal space (PVS) without entering the retropubic space (RPS). STUDY DESIGN: Eight patients undergoing vaginal paravaginal repair were enrolled. The paravaginal dissection was completed to the most cephalad portion of the PVS without entering the RPS. The apex of each PVS was stained with methylene blue. The RPS was entered, the ATFP visualized, and 4 sutures were placed along its length to be used for the repair. The perpendicular distance between each suture and the most cephalad area of stain was measured. RESULTS: The mean distance from the perceived to actual ATFP at each suture point (1-4) was 3.5 cm, 2.75 cm, 2.0 cm, and 0.91 cm, respectively. CONCLUSION: In these 8 cases, the RPS had to be entered to accurately identify the ATFP. The degree of error increases as the ischial spine is approached (P < .001). PMID- 15902183 TI - Evaluation of the role of pudendal nerve integrity in female sexual function using noninvasive techniques. AB - OBJECTIVE: Using quantitative sensory testing and a validated questionnaire, we investigated the role of pudendal nerve integrity in sexual function among women. STUDY DESIGN: Participants completed the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ). Vibratory and pressure thresholds were measured at the S2 dermatome reflecting pudendal nerve distribution. RESULTS: A total of 56 women enrolled; 29 (51.8%) were asymptomatic and 27 (48.2%) had 1 or more forms of female sexual dysfunction (total sexual dysfunction) including: desire disorder 16.1%, arousal disorder 26.8%, orgasmic disorder 25%, and pain disorder 12.5%. Age, parity, menopausal status, and body mass index were similar between groups. PISQ scores were lower in symptomatic subjects compared with controls (P < .001). Decreased tactile sensation was found at the clitoris for women with total sexual dysfunction, desire disorder, and arousal disorder. Women with arousal disorder also had decreased tactile sensation at the perineum. CONCLUSION: Pudendal nerve integrity may play a role in female sexual dysfunction. PMID- 15902184 TI - Perisurgical smoking cessation and reduction of postoperative complications. AB - OBJECTIVE: The purpose of this study was to determine if a perisurgical smoking cessation program reduces smoking-related postoperative complications in urogynecologic surgery. STUDY DESIGN: A review of patients that underwent pelvic reconstructive surgery from 1998 to 2003 was performed. All smokers underwent a perisurgical smoking cessation program of their choice for at least 1 month before surgery, and continued for 1 month after surgery. Complications unrelated to smoking (cystotomy, enterotomy, urethral obstruction, etc) were excluded in the smoking-potentiated complications. Problems considered to be potentiated by smoking were: wound, pulmonary, cardiac, and febrile morbidity. RESULTS: Eight hundred eighty-seven patients were included. There were 233 smoker cessation patients (SC) and 654 nonsmokers (NS). The total number of complications in the SC group was 61 (61/233, 26%) compared with 172 (172/654, 29%) in the NS group: (chi-square, P = .97). When looking at smoking-potentiated complications only, there were 34 (34/61, 56%) patients in the SC group and 90 (90/172, 52%) in the NS group (chi-square, P = .75). CONCLUSION: There are no differences in smoking potentiated complications between nonsmoking patients and patients who undergo a perisurgical smoking cessation program. PMID- 15902185 TI - Does vaginal closure force differ in the supine and standing positions? AB - OBJECTIVE: This study was undertaken to quantify resting vaginal closure force (VCF(REST)), maximum vaginal closure force (VCF(MAX)), and augmentation of vaginal closure force augmentation (VCF(AUG)) when supine and standing and to determine whether the change in intra-abdominal pressure associated with change in posture accounts for differences in VCF. STUDY DESIGN: Thirty-nine asymptomatic, continent women were recruited to determine, when supine and standing, the vaginal closure force (eg, the force closing the vagina in the mid sagittal plane) and bladder pressures at rest and at maximal voluntary contraction. VCF was measured with an instrumented vaginal speculum and bladder pressure was determined with a microtip catheter. VCF(REST) was the resting pelvic floor tone, and VCF(MAX) was the peak pelvic floor force during a maximal voluntary contraction. VCF(AUG) was the difference between VCF(MAX) and VCF(REST). T tests and Pearson correlation coefficients were used for analysis. RESULTS: VCF(REST) when supine was 3.6 +/- 0.8 N and when standing was 6.9 +/- 1.5 N--a 92% difference (P < .001). The VCF(MAX) when supine was 7.5 +/- 2.9 N and when standing was 10.1 +/- 2.4 N--a 35% difference (P < .001). Bladder pressure when supine (10.5 +/- 4.7 cm H2O) was significantly less (P < .001) than when standing (31.0 +/- 6.4 cm H2O). The differences in bladder pressure when either supine or standing did not correlate with the corresponding differences in VCF at rest or at maximal voluntary contraction. The supine VCF(AUG) of 3.9 +/- 2.7 N, was significantly greater than the standing VCF(AUG) of 3.3+/-1.9 N. CONCLUSION: With change in posture, vaginal closure force increases because of higher intra-abdominal pressure and greater resistance in the pelvic floor muscles. PMID- 15902186 TI - Extensive fever workup produces low yield in determining infectious etiology. AB - OBJECTIVE: The study was undertaken to evaluate the use of a fever workup in women undergoing benign gynecologic procedures. STUDY DESIGN: A retrospective chart review was performed at Jackson Memorial Hospital between 1994 and 2000. Information was abstracted from hospital and clinic records. Fever criteria was defined as 1 temperature equal to or greater than 101.5, or 2 equal to or greater than 100.4, at least 4 hours apart within a 24-hour period. Patients undergoing additional intraoperative procedures leading to increased febrile morbidity were excluded. Data abstracted included patient demographics, procedure, complications, antibiotic use, and extent of fever workup. Statistical analysis used was 2-sample t tests, Wilcoxon rank test, chi2 test, and multivariate logistic regression. Alpha level = .05. RESULTS: The charts of 505 patients were reviewed, and 147 patients met fever criteria. All patients underwent surgery for benign conditions, abdominal hysterectomy being the most common (90%). The study population was divided into 2 groups: the noninfectious group and infectious group. These groups were determined by wound infection, pelvic abscess, blood or urine culture, ultrasound, and chest roentgen. Both groups were found to be similar with respect to demographics, surgical procedures, and postoperative complications, with the exception of body mass index (28.4 vs 31.7) and length of hospital stay (3.9 vs 5.3). Results from fever workups included positive results blood cultures (9.7%), urine culture (18.8%), and chest roentgens (14%) in this study population. We found no association between positive urine analysis and urine culture. When comparing both groups, a statistically significant difference was found with regard to maximum temperature elevation, number of days febrile, and postoperative day of maximum temperature (P < .05). CONCLUSION: The extensive fever workup was not frequently positive in this study population. Its use and cost-effectiveness should be questioned. Therefore, the fever workup should be tailored to the individual patient. PMID- 15902187 TI - Efficacy of botulinum-A toxin in the treatment of detrusor overactivity incontinence: a prospective nonrandomized study. AB - OBJECTIVE: This study was undertaken to investigate the efficacy and safety of botulinum-A toxin (BTX-A) treatment for non-neurogenic detrusor overactivity incontinence. STUDY DESIGN: This prospective nonrandomized ongoing study was performed in a tertiary referral urogynecology department. In 26 women with urge incontinence and urodynamically demonstrated detrusor overactivity incontinence resistant to conventional treatment 100 units of BTX-A were injected into the detrusor muscle at 30 sites. Clinical and urodynamic evaluations and a quality of life assessment were performed at baseline and 4, 12, and 36 weeks after BTX-A treatment. RESULTS: Of 26 women, 14 were dry after 4 weeks, 13 of 20 women after 12 weeks, and 3 of 5 women after 36 weeks. Two women failed to respond. Two women were on self-catheterization temporarily. There were no other complications besides 9 urinary tract infections within the 51 follow-up visits. CONCLUSION: BTX-A treatment seems to be a safe and efficacious new treatment option for patients with detrusor overactivity incontinence. PMID- 15902188 TI - The effects of hormone replacement on the biomechanical properties of the uterosacral and round ligaments in the monkey model. AB - OBJECTIVE: The purpose of this study was to determine effects of ovariectomy (OVX) and conjugated equine estrogens plus medroxyprogesterone acetate (CEE/MPA), or ethinyl estradiol plus norethindrone acetate (EE/NA) on biomechanics of uterosacral (USL) and round (RL) ligaments in postmenopausal (PMP) monkeys. STUDY DESIGN: This was a randomized, triple blind, placebo-controlled study. OVX monkeys received 12 months no treatment (Pbo) (n = 19), CEE/MPA (n = 19), or EE/NA (n = 21). USL and RL step strains and stress-relaxation data were curve fitted, giving strain-dependent tensile modulus (TM) from 0% to 30%. RESULTS: (1) USL: TM for both treatment groups was greater than Pbo for strains from 0% to 12% (P < .04). (2) RL: TM for both treatment groups was smaller than Pbo for strains from 12% to 30% (P < .05). No differences were found between treatment regimens. CONCLUSION: CEE/MPA and EE/NA both affect functional biomechanical properties by increasing tensile stiffness in the USL and decreasing it in the RL. PMID- 15902189 TI - Laparoscopic and abdominal sacral colpopexies: a comparative cohort study. AB - OBJECTIVE: This study was undertaken to compare laparoscopic and open sacral colpopexies for efficacy and safety. STUDY DESIGN: Charts were reviewed for 56 patients who underwent laparoscopic sacral colpopexy and 61 patients who underwent open sacral colpopexy. Demographic and hospital data, complications, and follow-up visits were reviewed. RESULTS: Mean follow-up was 13.5 +/- 12.1 months and 15.7 +/- 18.1 months in the laparoscopic and open groups, respectively. Mean operating time was significantly greater in the laparoscopic versus open cohort, 269 +/- 65 minutes and 218 +/- 60 minutes, respectively (P < .0001). Estimated blood loss (172 +/- 166 mL vs 234 +/- 149 mL; P = .04) and hospital stay (1.8 +/- 1.0 days vs 4.0 +/- 1.8 days; P < .0001) were significantly less in the laparoscopic group than the open group. Complication and reoperation rates were similar. CONCLUSION: Laparoscopic and open sacral colpopexies have comparable clinical outcomes. Although laparoscopic sacral colpopexy requires longer operating time, hospital stay is significantly decreased. PMID- 15902191 TI - Vaginal wall stretching. PMID- 15902192 TI - Peer review is complicated. PMID- 15902194 TI - Cortical Fluoro-Jade staining and blunted adrenomedullary response to hypoglycemia after noncoma hypoglycemia in rats. AB - Intensive insulin therapy in patients with type 1 diabetes mellitus reduces long term complications; however, intensive therapy is also associated with a three fold increase in hypoglycemic episodes. The present study in conscious rats characterizes the physiologic and neuropathologic consequences of a single episode of moderate hypoglycemia. In this model, intravenous insulin is used to reduce plasma glucose to 30 to 35 mg/dL for 75 mins. This single hypoglycemic insult acutely induces hypoglycemia-associated autonomic failure (HAAF), with epinephrine responses to hypoglycemia reduced more than 36% from control. Neuropathology after this insult includes the appearance of dying cells, assessed with the marker Fluoro-jade B (FJ). After hypoglycemic insult, FJ+ cells were consistently seen in subdivisions of the medial prefrontal cortex, the orbital cortex, and the piriform cortex. There was a significant correlation between depth of hypoglycemia and number of FJ+ cells, suggesting that there is a critical threshold below which vulnerable cells begin to die. These data suggest that there is a population of cells that are vulnerable to moderate levels of hypoglycemia commonly experienced by patients with insulin-treated diabetes. These cells, which may be neurons, are primarily found in cortical regions implicated in visceral perception and autonomic control, raising the possibility that their loss contributes to clinically reported deficits in autonomic and perceptual responses to hypoglycemia. PMID- 15902195 TI - VEGF-induced BBB permeability is associated with an MMP-9 activity increase in cerebral ischemia: both effects decreased by Ang-1. AB - After cerebral ischemia, angiogenesis, by supplying for the deficient perfusion, may be a beneficial process for limiting neuronal death and promoting tissue repair. In this study, we showed that the combination of Ang-1 and vascular endothelial growth factor (VEGF) provides a more adapted therapeutic strategy than the use of VEGF alone. Indeed, we showed on a focal ischemia model that an early administration of VEGF exacerbates ischemic damage, because of its effects on blood-brain barrier (BBB) permeability. In contrast, a coapplication of Ang-1 and VEGF leads to a significant reduction of the ischemic and edema volumes by 50% and 42%, respectively, in comparison with VEGF-treated mice. We proposed that Ang-1 blocks the BBB permeability effect of VEGF in association with a modulation of matrix metalloproteinase (MMP) activity. Indeed, we showed on both ischemic in vivo and BBB in vitro models that VEGF enhances BBB damage and MMP-9 activity and that Ang-1 counteracts both effects. However, we also showed a synergic angiogenic effect of Ang-1 and VEGF in the brain. Taken together, these results allow to propose that, in cerebral ischemia, the combination of Ang-1 and VEGF could be used early to promote the formation of mature neovessels without inducing side effects on BBB permeability. PMID- 15902196 TI - Effect of targeted deletion of the heme oxygenase-2 gene on hemoglobin toxicity in the striatum. AB - The heme oxygenase (HO) enzymes catalyze the rate-limiting step in the breakdown of heme to iron, carbon monoxide, and biliverdin. A prior cell culture study demonstrated that deletion of HO-2, the isoform constitutively expressed in neurons, attenuated hemoglobin (Hb) neurotoxicity. The present study tested the hypothesis that HO-2 gene deletion is cytoprotective in a model of Hb toxicity in vivo. Stereotactic injection of 6 microL stroma-free Hb (SFHb) into the striatum significantly increased protein oxidation in wild-type mice at 24 to 72 h, as detected by an assay for carbonyl groups. At 72 h, carbonylation was increased 2.5-fold compared with that in the contralateral striatum. In HO-2 knockout mice, protein oxidation was not increased at 24 h, and was increased by only 1.7-fold at 72 h. Similarly, striatal lipid peroxidation, as detected by the malondialdehyde assay, was significantly greater in the SFHb-injected striata of wild-type mice than in knockout mice. Striatal cell viability, determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, was 45.0%+/ 6.3% of that in contralateral striata in wild-type mice at 72 h; it was increased to 85%+/-8% in knockouts. Heme oxygenase-2 gene deletion did not alter weight loss or mortality after SFHb injection. Baseline striatal HO-1 expression was similar in knockout and wild-type mice; induction after SFHb injection occurred more rapidly in the latter. These results suggest that HO-2 gene deletion protects striatal cells from the oxidative toxicity of Hb in vivo. Pharmacologic or genetic strategies that target HO-2 may be beneficial after central nervous system hemorrhage, and warrant further investigation. PMID- 15902197 TI - Heat acclimation increases hypoxia-inducible factor 1alpha and erythropoietin receptor expression: implication for neuroprotection after closed head injury in mice. AB - Experimental evidence indicates that long-term exposure to moderately high ambient temperature (heat acclimation, HA) mediates cross-tolerance to various types of subsequently applied stress. The transcriptional activator hypoxia inducible factor 1 (HIF-1) has been implicated in playing a critical role in HA. It also regulates the expression of Erythropoietin (Epo), whose neuroprotective effects have been shown in a variety of brain injuries. The aim of the present study was to examine whether HA exerts a beneficial effect on the outcome of closed head injury (CHI) in mice and to explore the possible involvement of HIF-1 and Epo in this process. Heat acclimated mice and matched normothermic controls were subjected to CHI or sham surgery. Postinjury motor and cognitive parameters of acclimated mice were compared with those of controls. Mice were killed at various time points after injury or sham surgery and brain levels of HIF-1alpha, the inducible subunit of HIF-1, Epo, and the specific erythropoietin receptor (EpoR) were analyzed by Western immunoblotting. Motor and cognitive functions of acclimated mice were significantly better than those of controls. Heat acclimation was found to induce a significant increase in expression of nuclear HIF-1alpha and EpoR. The EpoR/Epo ratio was also significantly higher in acclimated mice as compared with controls. Nuclear HIF-1alpha and EpoR were higher in the acclimated group at 4 h after injury as well. The improved outcome of acclimated mice taken together with the basal and postinjury upregulation of the examined proteins suggests the involvement of this pathway in HA-induced neuroprotection. PMID- 15902198 TI - Serum ferritin in stroke: a marker of increased body iron stores or stroke severity? AB - To evaluate the effect of body iron stores on the vulnerability of the brain to ischemia, a focal permanent brain ischemia was induced by photothrombotic occlusion of cortical vessels in rats with or without chronic treatment with iron dextran (25 mg iron/kg, every other day for 20 days, intraperitoneally). Iron dextran induced systemic iron overload as evidenced by high ferritin (Ft) ( x 5) and total iron levels ( x 3) in serum as well as increased Ft expression in the liver and heart. Conversely, neither serum free iron levels nor Ft expression in the brain were changed by iron dextran. Finally, infarct volume was not modified by iron dextran. In addition, induction of ischemia in rats treated with FeCl(3) (560 microg iron/kg, intravenously) as a means of increasing serum free iron levels during the ischemic period did not enlarge infarct volume. We then explored the effect of brain ischemia itself on serum Ft by measuring serum Ft before and after induction of brain ischemic insults with different neurologic outcomes in rats (brain embolization with microspheres, photothrombotic occlusion of cortical vessels, four-vessel occlusion). Serum Ft levels were found higher at day 1 after ischemia than before ischemia only in rats subjected to the most severe insult (brain embolization). In conclusion, our study showed that increased body iron stores do not increase the vulnerability of the brain to ischemia and that brain ischemia, if severe, results in the elevation of serum Ft levels. PMID- 15902199 TI - Novel surrogate markers for acute brain damage: cerebrospinal fluid levels corrrelate with severity of ischemic neurodegeneration in the rat. AB - Previously, we identified proteins released from degenerating cultured cortical neurons as novel cerebrospinal fluid (CSF) markers for acute brain injury in the rat. Here, we investigate relationships between CSF changes in these novel markers and the severity of acute ischemic brain injury. Rats underwent sham surgery or 3,6,8, or 10 mins of transient global forebrain ischemia. At 48 h after insult, CSF levels of 14-3-3beta, 14-3-3zeta, and calpain cleavage products of alpha-spectrin and tau were quantified. Regional acute neurodegeneration was assessed by Fluoro-Jade and silver impregnation staining, and confirmed by immunohistochemical detection of the activation of calpain and caspase, cysteine proteases involved in neurodegenerative signaling. Ischemic neurodegeneration and activation of at least one cysteine protease were observed in the hippocampal CA1 sector, dentate hilus, caudate nucleus, parietal cortex, thalamus, and inferior colliculus. As expected, the total number of degenerating cells increased as a function of ischemia duration. Cerebrospinal fluid levels of the four marker proteins increased markedly after ischemia, and rose in proportion with its duration. Irrespective of the length of ischemia, CSF levels of the neuron enriched proteins 14-3-3beta and calpain-cleaved tau correlated significantly with the magnitude of acute ischemic neurodegeneration. Additionally, CSF levels of the two proteins correlated with one another. These results show that certain proteins released from degenerating neurons are CSF markers for brain injury in the rat whose levels reflect the severity of acute ischemic neurodegeneration. Measurement of 14-3-3beta and calpain-cleaved tau may be useful for the minimally invasive diagnosis, prognosis, and therapeutic evaluation of acute brain damage. PMID- 15902200 TI - Differential profile of Nix upregulation and translocation during hypoxia/ischaemia in vivo versus in vitro. AB - Nix, a hypoxia-sensitive member of the Bcl-2 family, is upregulated at the mRNA level during hypoxia through induction of a hypoxia-inducible factor-1 alpha (HIF 1 alpha) response element in its promoter sequence. However, the mechanism(s) regulating Nix protein activation remain unclear. The present studies examine Nix protein expression and subcellular distribution in response to hypoxic stimuli in vivo and in culture and to two disparate apoptotic stimuli in vitro. Upregulation and translocation of Nix (by day 5) in hypoxic/serum-deprived CHO-K1 cells, was preceded by Bax activation (by day 4) and caspase-3 processing (by day 2), suggesting that initiation of cell death in vitro is a Nix-independent event. In contrast, an early Nix response (upregulation and translocation to the mitochondria) was observed after 6 h of middle cerebral artery occlusion in the rat. Nix translocation was observed in the ipsilateral cortex and striatum before other histological (infarct development, neuronal loss, apoptotic body formation) or biochemical (Bax activation or caspase-3 cleavage) markers of damage were detected. While fundamental differences between hypoxia/ischaemia in culture and in vivo likely explain the different temporal profiles of Nix, Bax, and caspase-3 activation observed, these studies show that like Bax, mitochondrial accumulation is a common event during Nix activation. These are the first studies to show upregulation and translocation of Nix in the ischaemic brain and suggest Nix to be a novel therapeutic target in ischaemic research. Moreover, Nix upregulation in staurosporine-treated SH-SY5Y cells and dexamethasone-treated A1.1 cells supports a more generalized role for Nix in apoptotic cell death. PMID- 15902201 TI - Metabolism is normal in astrocytes in chronically epileptic rats: a (13)C NMR study of neuronal-glial interactions in a model of temporal lobe epilepsy. AB - The aim of the present work was to study potential disturbances in metabolism and interactions between neurons and glia in the lithium-pilocarpine model of temporal lobe epilepsy. Rats chronically epileptic for 1 month received [1 (13)C]glucose, a substrate for neurons and astrocytes, and [1,2-(13)C]acetate, a substrate for astrocytes only. Analyses of extracts from cerebral cortex, cerebellum, and hippocampal formation (hippocampus, amygdala, entorhinal, and piriform cortices) were performed using (13)C and (1)H nuclear magnetic resonance spectroscopy and HPLC. In the hippocampal formation of epileptic rats, levels of glutamate, aspartate, N-acetyl aspartate, adenosine triphosphate plus adenosine diphosphate and glutathione were decreased. In all regions studied, labeling from [1,2-(13)C]acetate was similar in control and epileptic rats, indicating normal astrocytic metabolism. However, labeling of glutamate, GABA, aspartate, and alanine from [1-(13)C]glucose was decreased in all areas possibly reflecting neuronal loss. The labeling of glutamine from [1-(13)C]glucose was decreased in cerebral cortex and cerebellum and unchanged in hippocampal formation. In conclusion, no changes were detected in glial-neuronal interactions in the hippocampal formation while in cortex and cerebellum the flow of glutamate to astrocytes was decreased, indicating a disturbed glutamate-glutamine cycle. This is, to our knowledge, the first study showing that metabolic disturbances are confined to neurons inside the epileptic circuit. PMID- 15902202 TI - Peripheral is central to the question. PMID- 15902206 TI - Ephrin signalling controls brain size by regulating apoptosis of neural progenitors. AB - Mechanisms controlling brain size include the regulation of neural progenitor cell proliferation, differentiation, survival and migration. Here we show that ephrin-A/EphA receptor signalling plays a key role in controlling the size of the mouse cerebral cortex by regulating cortical progenitor cell apoptosis. In vivo gain of EphA receptor function, achieved through ectopic expression of ephrin-A5 in early cortical progenitors expressing EphA7, caused a transient wave of neural progenitor cell apoptosis, resulting in premature depletion of progenitors and a subsequent dramatic decrease in cortical size. In vitro treatment with soluble ephrin-A ligands similarly induced the rapid death of cultured dissociated cortical progenitors in a caspase-3-dependent manner, thereby confirming a direct effect of ephrin/Eph signalling on apoptotic cascades. Conversely, in vivo loss of EphA function, achieved through EphA7 gene disruption, caused a reduction in apoptosis occurring normally in forebrain neural progenitors, resulting in an increase in cortical size and, in extreme cases, exencephalic forebrain overgrowth. Together, these results identify ephrin/Eph signalling as a physiological trigger for apoptosis that can alter brain size and shape by regulating the number of neural progenitors. PMID- 15902207 TI - Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor. AB - Changes in membrane potential affect ion channels and transporters, which then alter intracellular chemical conditions. Other signalling pathways coupled to membrane potential have been suggested but their underlying mechanisms are unknown. Here we describe a novel protein from the ascidian Ciona intestinalis that has a transmembrane voltage-sensing domain homologous to the S1-S4 segments of voltage-gated channels and a cytoplasmic domain similar to phosphatase and tensin homologue. This protein, named C. intestinalis voltage-sensor-containing phosphatase (Ci-VSP), displays channel-like 'gating' currents and directly translates changes in membrane potential into the turnover of phosphoinositides. The activity of the phosphoinositide phosphatase in Ci-VSP is tuned within a physiological range of membrane potential. Immunocytochemical studies show that Ci-VSP is expressed in Ciona sperm tail membranes, indicating a possible role in sperm function or morphology. Our data demonstrate that voltage sensing can function beyond channel proteins and thus more ubiquitously than previously realized. PMID- 15902208 TI - An inhibitor of Bcl-2 family proteins induces regression of solid tumours. AB - Proteins in the Bcl-2 family are central regulators of programmed cell death, and members that inhibit apoptosis, such as Bcl-X(L) and Bcl-2, are overexpressed in many cancers and contribute to tumour initiation, progression and resistance to therapy. Bcl-X(L) expression correlates with chemo-resistance of tumour cell lines, and reductions in Bcl-2 increase sensitivity to anticancer drugs and enhance in vivo survival. The development of inhibitors of these proteins as potential anti-cancer therapeutics has been previously explored, but obtaining potent small-molecule inhibitors has proved difficult owing to the necessity of targeting a protein-protein interaction. Here, using nuclear magnetic resonance (NMR)-based screening, parallel synthesis and structure-based design, we have discovered ABT-737, a small-molecule inhibitor of the anti-apoptotic proteins Bcl 2, Bcl-X(L) and Bcl-w, with an affinity two to three orders of magnitude more potent than previously reported compounds. Mechanistic studies reveal that ABT 737 does not directly initiate the apoptotic process, but enhances the effects of death signals, displaying synergistic cytotoxicity with chemotherapeutics and radiation. ABT-737 exhibits single-agent-mechanism-based killing of cells from lymphoma and small-cell lung carcinoma lines, as well as primary patient-derived cells, and in animal models, ABT-737 improves survival, causes regression of established tumours, and produces cures in a high percentage of the mice. PMID- 15902209 TI - Iran's long march. PMID- 15902210 TI - Running out of juice. PMID- 15902211 TI - Policing integrity. PMID- 15902213 TI - Large genomic differences explain our little quirks. PMID- 15902214 TI - Planet hunters lose out to Hubble rescue. PMID- 15902216 TI - Ethicists urge caution over emotive power of brain scans. PMID- 15902217 TI - US aid agency grilled over malaria funds. PMID- 15902218 TI - Snapshot: Eureka moment as X-rays slice through forgery. PMID- 15902219 TI - Special report: taking on the cheats. PMID- 15902221 TI - Iranian neuroscience: the brains trust of Tehran. PMID- 15902222 TI - Mars exploration: going underground. PMID- 15902223 TI - Stem-cell niches: it's the ecology, stupid! PMID- 15902224 TI - Licensing fees slow advance of stem cells. PMID- 15902227 TI - Seeking evidence of God's work undermines faith. PMID- 15902228 TI - Evolution is a short-order cook, not a watchmaker. PMID- 15902229 TI - Teaching about ID helps students see its flaws. PMID- 15902230 TI - When science meets religion in the classroom. PMID- 15902231 TI - Solidarity with the oppressed flat-Earthers. PMID- 15902232 TI - Intelligent design or intellectual laziness? PMID- 15902238 TI - Capturing chaos. PMID- 15902233 TI - Leave well alone and stick to teaching what you know. PMID- 15902239 TI - Evolutionary biology: geography and skin colour. PMID- 15902240 TI - Earthquakes: future shock in California. PMID- 15902241 TI - Molecular motors: kinesin steps back. PMID- 15902242 TI - Particle physics: vanishing pentaquarks. PMID- 15902243 TI - Neuroscience: plasticity and its limits. PMID- 15902244 TI - Timekeeping: light-insensitive optical clock. PMID- 15902245 TI - Obituary: Saunders Mac Lane (1909-2005). PMID- 15902246 TI - Psychology: red enhances human performance in contests. AB - Red coloration is a sexually selected, testosterone-dependent signal of male quality in a variety of animals, and in some non-human species a male's dominance can be experimentally increased by attaching artificial red stimuli. Here we show that a similar effect can influence the outcome of physical contests in humans- across a range of sports, we find that wearing red is consistently associated with a higher probability of winning. These results indicate not only that sexual selection may have influenced the evolution of human response to colours, but also that the colour of sportswear needs to be taken into account to ensure a level playing field in sport. PMID- 15902247 TI - Three-dimensional deformation caused by the Bam, Iran, earthquake and the origin of shallow slip deficit. AB - Our understanding of the earthquake process requires detailed insights into how the tectonic stresses are accumulated and released on seismogenic faults. We derive the full vector displacement field due to the Bam, Iran, earthquake of moment magnitude 6.5 using radar data from the Envisat satellite of the European Space Agency. Analysis of surface deformation indicates that most of the seismic moment release along the 20-km-long strike-slip rupture occurred at a shallow depth of 4-5 km, yet the rupture did not break the surface. The Bam event may therefore represent an end-member case of the 'shallow slip deficit' model, which postulates that coseismic slip in the uppermost crust is systematically less than that at seismogenic depths (4-10 km). The InSAR-derived surface displacement data from the Bam and other large shallow earthquakes suggest that the uppermost section of the seismogenic crust around young and developing faults may undergo a distributed failure in the interseismic period, thereby accumulating little elastic strain. PMID- 15902248 TI - Lack of long-term cortical reorganization after macaque retinal lesions. AB - Several aspects of cortical organization are thought to remain plastic into adulthood, allowing cortical sensorimotor maps to be modified continuously by experience. This dynamic nature of cortical circuitry is important for learning, as well as for repair after injury to the nervous system. Electrophysiology studies suggest that adult macaque primary visual cortex (V1) undergoes large scale reorganization within a few months after retinal lesioning, but this issue has not been conclusively settled. Here we applied the technique of functional magnetic resonance imaging (fMRI) to detect changes in the cortical topography of macaque area V1 after binocular retinal lesions. fMRI allows non-invasive, in vivo, long-term monitoring of cortical activity with a wide field of view, sampling signals from multiple neurons per unit cortical area. We show that, in contrast with previous studies, adult macaque V1 does not approach normal responsivity during 7.5 months of follow-up after retinal lesions, and its topography does not change. Electrophysiology experiments corroborated the fMRI results. This indicates that adult macaque V1 has limited potential for reorganization in the months following retinal injury. PMID- 15902249 TI - Mechanics of the kinesin step. AB - Kinesin is a molecular walking machine that organizes cells by hauling packets of components directionally along microtubules. The physical mechanism that impels directional stepping is uncertain. We show here that, under very high backward loads, the intrinsic directional bias in kinesin stepping can be reversed such that the motor walks sustainedly backwards in a previously undescribed mode of ATP-dependent backward processivity. We find that both forward and backward 8-nm steps occur on the microsecond timescale and that both occur without mechanical substeps on this timescale. The data suggest an underlying mechanism in which, once ATP has bound to the microtubule-attached head, the other head undergoes a diffusional search for its next site, the outcome of which can be biased by an applied load. PMID- 15902250 TI - Detection and imaging of atmospheric radio flashes from cosmic ray air showers. AB - The nature of ultrahigh-energy cosmic rays (UHECRs) at energies >10(20) eV remains a mystery. They are likely to be of extragalactic origin, but should be absorbed within approximately 50 Mpc through interactions with the cosmic microwave background. As there are no sufficiently powerful accelerators within this distance from the Galaxy, explanations for UHECRs range from unusual astrophysical sources to exotic string physics. Also unclear is whether UHECRs consist of protons, heavy nuclei, neutrinos or gamma-rays. To resolve these questions, larger detectors with higher duty cycles and which combine multiple detection techniques are needed. Radio emission from UHECRs, on the other hand, is unaffected by attenuation, has a high duty cycle, gives calorimetric measurements and provides high directional accuracy. Here we report the detection of radio flashes from cosmic-ray air showers using low-cost digital radio receivers. We show that the radiation can be understood in terms of the geosynchrotron effect. Our results show that it should be possible to determine the nature and composition of UHECRs with combined radio and particle detectors, and to detect the ultrahigh-energy neutrinos expected from flavour mixing. PMID- 15902251 TI - CO self-shielding as the origin of oxygen isotope anomalies in the early solar nebula. AB - The abundances of oxygen isotopes in the most refractory mineral phases (calcium aluminium-rich inclusions, CAIs) in meteorites have hitherto defied explanation. Most processes fractionate isotopes by nuclear mass; that is, 18O is twice as fractionated as 17O, relative to 16O. In CAIs 17O and 18O are nearly equally fractionated, implying a fundamentally different mechanism. The CAI data were originally interpreted as evidence for supernova input of pure 16O into the solar nebula, but the lack of a similar isotope trend in other elements argues against this explanation. A symmetry-dependent fractionation mechanism may have occurred in the inner solar nebula, but experimental evidence is lacking. Isotope selective photodissociation of CO in the innermost solar nebula might explain the CAI data, but the high temperatures in this region would have rapidly erased the signature. Here we report time-dependent calculations of CO photodissociation in the cooler surface region of a turbulent nebula. If the surface were irradiated by a far-ultraviolet flux approximately 10(3) times that of the local interstellar medium (for example, owing to an O or B star within approximately 1 pc of the protosun), then substantial fractionation of the oxygen isotopes was possible on a timescale of approximately 10(5) years. We predict that similarly irradiated protoplanetary disks will have H2O enriched in 17O and 18O by several tens of per cent relative to CO. PMID- 15902252 TI - An optical lattice clock. AB - The precision measurement of time and frequency is a prerequisite not only for fundamental science but also for technologies that support broadband communication networks and navigation with global positioning systems (GPS). The SI second is currently realized by the microwave transition of Cs atoms with a fractional uncertainty of 10(-15) (ref. 1). Thanks to the optical frequency comb technique, which established a coherent link between optical and radio frequencies, optical clocks have attracted increasing interest as regards future atomic clocks with superior precision. To date, single trapped ions and ultracold neutral atoms in free fall have shown record high performance that is approaching that of the best Cs fountain clocks. Here we report a different approach, in which atoms trapped in an optical lattice serve as quantum references. The 'optical lattice clock' demonstrates a linewidth one order of magnitude narrower than that observed for neutral-atom optical clocks, and its stability is better than that of single-ion clocks. The transition frequency for the Sr lattice clock is 429,228,004,229,952(15) Hz, as determined by an optical frequency comb referenced to the SI second. PMID- 15902253 TI - Micrometre-scale silicon electro-optic modulator. AB - Metal interconnections are expected to become the limiting factor for the performance of electronic systems as transistors continue to shrink in size. Replacing them by optical interconnections, at different levels ranging from rack to-rack down to chip-to-chip and intra-chip interconnections, could provide the low power dissipation, low latencies and high bandwidths that are needed. The implementation of optical interconnections relies on the development of micro optical devices that are integrated with the microelectronics on chips. Recent demonstrations of silicon low-loss waveguides, light emitters, amplifiers and lasers approach this goal, but a small silicon electro-optic modulator with a size small enough for chip-scale integration has not yet been demonstrated. Here we experimentally demonstrate a high-speed electro-optical modulator in compact silicon structures. The modulator is based on a resonant light-confining structure that enhances the sensitivity of light to small changes in refractive index of the silicon and also enables high-speed operation. The modulator is 12 micrometres in diameter, three orders of magnitude smaller than previously demonstrated. Electro-optic modulators are one of the most critical components in optoelectronic integration, and decreasing their size may enable novel chip architectures. PMID- 15902254 TI - Real-time forecasts of tomorrow's earthquakes in California. AB - Despite a lack of reliable deterministic earthquake precursors, seismologists have significant predictive information about earthquake activity from an increasingly accurate understanding of the clustering properties of earthquakes. In the past 15 years, time-dependent earthquake probabilities based on a generic short-term clustering model have been made publicly available in near-real time during major earthquake sequences. These forecasts describe the probability and number of events that are, on average, likely to occur following a mainshock of a given magnitude, but are not tailored to the particular sequence at hand and contain no information about the likely locations of the aftershocks. Our model builds upon the basic principles of this generic forecast model in two ways: it recasts the forecast in terms of the probability of strong ground shaking, and it combines an existing time-independent earthquake occurrence model based on fault data and historical earthquakes with increasingly complex models describing the local time-dependent earthquake clustering. The result is a time-dependent map showing the probability of strong shaking anywhere in California within the next 24 hours. The seismic hazard modelling approach we describe provides a better understanding of time-dependent earthquake hazard, and increases its usefulness for the public, emergency planners and the media. PMID- 15902255 TI - Direct dating of Early Upper Palaeolithic human remains from Mladec. AB - The human fossil assemblage from the Mladec Caves in Moravia (Czech Republic) has been considered to derive from a middle or later phase of the Central European Aurignacian period on the basis of archaeological remains (a few stone artefacts and organic items such as bone points, awls, perforated teeth), despite questions of association between the human fossils and the archaeological materials and concerning the chronological implications of the limited archaeological remains. The morphological variability in the human assemblage, the presence of apparently archaic features in some specimens, and the assumed early date of the remains have made this fossil assemblage pivotal in assessments of modern human emergence within Europe. We present here the first successful direct accelerator mass spectrometry radiocarbon dating of five representative human fossils from the site. We selected sample materials from teeth and from one bone for 14C dating. The four tooth samples yielded uncalibrated ages of approximately 31,000 14C years before present, and the bone sample (an ulna) provided an uncertain more recent age. These data are sufficient to confirm that the Mladec human assemblage is the oldest cranial, dental and postcranial assemblage of early modern humans in Europe and is therefore central to discussions of modern human emergence in the northwestern Old World and the fate of the Neanderthals. PMID- 15902256 TI - Distinguishing random environmental fluctuations from ecological catastrophes for the North Pacific Ocean. AB - The prospect of rapid dynamic changes in the environment is a pressing concern that has profound management and public policy implications. Worries over sudden climate change and irreversible changes in ecosystems are rooted in the potential that nonlinear systems have for complex and 'pathological' behaviours. Nonlinear behaviours have been shown in model systems and in some natural systems, but their occurrence in large-scale marine environments remains controversial. Here we show that time series observations of key physical variables for the North Pacific Ocean that seem to show these behaviours are not deterministically nonlinear, and are best described as linear stochastic. In contrast, we find that time series for biological variables having similar properties exhibit a low dimensional nonlinear signature. To our knowledge, this is the first direct test for nonlinearity in large-scale physical and biological data for the marine environment. These results address a continuing debate over the origin of rapid shifts in certain key marine observations as coming from essentially stochastic processes or from dominant nonlinear mechanisms. Our measurements suggest that large-scale marine ecosystems are dynamically nonlinear, and as such have the capacity for dramatic change in response to stochastic fluctuations in basin scale physical states. PMID- 15902257 TI - Sustained firing in auditory cortex evoked by preferred stimuli. AB - It has been well documented that neurons in the auditory cortex of anaesthetized animals generally display transient responses to acoustic stimulation, and typically respond to a brief stimulus with one or fewer action potentials. The number of action potentials evoked by each stimulus usually does not increase with increasing stimulus duration. Such observations have long puzzled researchers across disciplines and raised serious questions regarding the role of the auditory cortex in encoding ongoing acoustic signals. Contrary to these long held views, here we show that single neurons in both primary (area A1) and lateral belt areas of the auditory cortex of awake marmoset monkeys (Callithrix jacchus) are capable of firing in a sustained manner over a prolonged period of time, especially when they are driven by their preferred stimuli. In contrast, responses become more transient or phasic when auditory cortex neurons respond to non-preferred stimuli. These findings suggest that when the auditory cortex is stimulated by a sound, a particular population of neurons fire maximally throughout the duration of the sound. Responses of other, less optimally driven neurons fade away quickly after stimulus onset. This results in a selective representation of the sound across both neuronal population and time. PMID- 15902258 TI - Regulation of PDGF signalling and vascular remodelling by peroxiredoxin II. AB - Platelet-derived growth factor (PDGF) is a potent mitogenic and migratory factor that regulates the tyrosine phosphorylation of a variety of signalling proteins via intracellular production of H2O2 (refs 1, 2-3). Mammalian 2-Cys peroxiredoxin type II (Prx II; gene symbol Prdx2) is a cellular peroxidase that eliminates endogenous H2O2 produced in response to growth factors such as PDGF and epidermal growth factor; however, its involvement in growth factor signalling is largely unknown. Here we show that Prx II is a negative regulator of PDGF signalling. Prx II deficiency results in increased production of H2O2, enhanced activation of PDGF receptor (PDGFR) and phospholipase Cgamma1, and subsequently increased cell proliferation and migration in response to PDGF. These responses are suppressed by expression of wild-type Prx II, but not an inactive mutant. Notably, Prx II is recruited to PDGFR upon PDGF stimulation, and suppresses protein tyrosine phosphatase inactivation. Prx II also leads to the suppression of PDGFR activation in primary culture and a murine restenosis model, including PDGF dependent neointimal thickening of vascular smooth muscle cells. These results demonstrate a localized role for endogenous H2O2 in PDGF signalling, and indicate a biological function of Prx II in cardiovascular disease. PMID- 15902259 TI - The Mesp2 transcription factor establishes segmental borders by suppressing Notch activity. AB - The serially segmented (metameric) structures of vertebrates are based on somites that are periodically formed during embryogenesis. A 'clock and wavefront' model has been proposed to explain the underlying mechanism of somite formation, in which the periodicity is generated by oscillation of Notch components (the clock) in the posterior pre-somitic mesoderm (PSM). This temporal periodicity is then translated into the segmental units in the 'wavefront'. The wavefront is thought to exist in the anterior PSM and progress backwards at a constant rate; however, there has been no direct evidence as to whether the levels of Notch activity really oscillate and how such oscillation is translated into a segmental pattern in the anterior PSM. Here, we have visualized endogenous levels of Notch1 activity in mice, showing that it oscillates in the posterior PSM but is arrested in the anterior PSM. Somite boundaries formed at the interface between Notch1 activated and -repressed domains. Genetic and biochemical studies indicate that this interface is generated by suppression of Notch activity by mesoderm posterior 2 (Mesp2) through induction of the lunatic fringe gene (Lfng). We propose that the oscillation of Notch activity is arrested and translated in the wavefront by Mesp2. PMID- 15902260 TI - The Ter mutation in the dead end gene causes germ cell loss and testicular germ cell tumours. AB - In mice, the Ter mutation causes primordial germ cell (PGC) loss in all genetic backgrounds. Ter is also a potent modifier of spontaneous testicular germ cell tumour (TGCT) susceptibility in the 129 family of inbred strains, and markedly increases TGCT incidence in 129-Ter/Ter males. In 129-Ter/Ter mice, some of the remaining PGCs transform into undifferentiated pluripotent embryonal carcinoma cells, and after birth differentiate into various cells and tissues that compose TGCTs. Here, we report the positional cloning of Ter, revealing a point mutation that introduces a termination codon in the mouse orthologue (Dnd1) of the zebrafish dead end (dnd) gene. PGC deficiency is corrected both with bacterial artificial chromosomes that contain Dnd1 and with a Dnd1-encoding transgene. Dnd1 is expressed in fetal gonads during the critical period when TGCTs originate. DND1 has an RNA recognition motif and is most similar to the apobec complementation factor, a component of the cytidine to uridine RNA-editing complex. These results suggest that Ter may adversely affect essential aspects of RNA biology during PGC development. DND1 is the first protein known to have an RNA recognition motif directly implicated as a heritable cause of spontaneous tumorigenesis. TGCT development in the 129-Ter mouse strain models paediatric TGCT in humans. This work will have important implications for our understanding of the genetic control of TGCT pathogenesis and PGC biology. PMID- 15902261 TI - Non-equilibration of hydrostatic pressure in blebbing cells. AB - Current models for protrusive motility in animal cells focus on cytoskeleton based mechanisms, where localized protrusion is driven by local regulation of actin biochemistry. In plants and fungi, protrusion is driven primarily by hydrostatic pressure. For hydrostatic pressure to drive localized protrusion in animal cells, it would have to be locally regulated, but current models treating cytoplasm as an incompressible viscoelastic continuum or viscous liquid require that hydrostatic pressure equilibrates essentially instantaneously over the whole cell. Here, we use cell blebs as reporters of local pressure in the cytoplasm. When we locally perfuse blebbing cells with cortex-relaxing drugs to dissipate pressure on one side, blebbing continues on the untreated side, implying non equilibration of pressure on scales of approximately 10 microm and 10 s. We can account for localization of pressure by considering the cytoplasm as a contractile, elastic network infiltrated by cytosol. Motion of the fluid relative to the network generates spatially heterogeneous transients in the pressure field, and can be described in the framework of poroelasticity. PMID- 15902262 TI - DNA synthesis provides the driving force to accelerate DNA unwinding by a helicase. AB - Helicases are molecular motors that use the energy of nucleoside 5'-triphosphate (NTP) hydrolysis to translocate along a nucleic acid strand and catalyse reactions such as DNA unwinding. The ring-shaped helicase of bacteriophage T7 translocates along single-stranded (ss)DNA at a speed of 130 bases per second; however, T7 helicase slows down nearly tenfold when unwinding the strands of duplex DNA. Here, we report that T7 DNA polymerase, which is unable to catalyse strand displacement DNA synthesis by itself, can increase the unwinding rate to 114 base pairs per second, bringing the helicase up to similar speeds compared to its translocation along ssDNA. The helicase rate of stimulation depends upon the DNA synthesis rate and does not rely on specific interactions between T7 DNA polymerase and the carboxy-terminal residues of T7 helicase. Efficient duplex DNA synthesis is achieved only by the combined action of the helicase and polymerase. The strand displacement DNA synthesis by the DNA polymerase depends on the unwinding activity of the helicase, which provides ssDNA template. The rapid trapping of the ssDNA bases by the DNA synthesis activity of the polymerase in turn drives the helicase to move forward through duplex DNA at speeds similar to those observed along ssDNA. PMID- 15902264 TI - To plan...or not to plan? PMID- 15902263 TI - Structure of the zinc-binding domain of an essential component of the hepatitis C virus replicase. AB - Hepatitis C virus (HCV) is a human pathogen affecting nearly 3% of the world's population. Chronic infections can lead to cirrhosis and liver cancer. The RNA replication machine of HCV is a multi-subunit membrane-associated complex. The non-structural protein NS5A is an active component of HCV replicase, as well as a pivotal regulator of replication and a modulator of cellular processes ranging from innate immunity to dysregulated cell growth. NS5A is a large phosphoprotein (56-58 kDa) with an amphipathic alpha-helix at its amino terminus that promotes membrane association. After this helix region, NS5A is organized into three domains. The N-terminal domain (domain I) coordinates a single zinc atom per protein molecule. Mutations disrupting either the membrane anchor or zinc binding of NS5A are lethal for RNA replication. However, probing the role of NS5A in replication has been hampered by a lack of structural information about this multifunctional protein. Here we report the structure of NS5A domain I at 2.5-A resolution, which contains a novel fold, a new zinc-coordination motif and a disulphide bond. We use molecular surface analysis to suggest the location of protein-, RNA- and membrane-interaction sites. PMID- 15902266 TI - Recruiters and academia. Lessons learned from a denied grant. Zen and the art of grant application. PMID- 15902269 TI - C --> T mutagenesis and gamma-radiation sensitivity due to deficiency in the Smug1 and Ung DNA glycosylases. AB - The most common genetic change in aerobic organisms is a C:G to T:A mutation. C - > T transitions can arise through spontaneous hydrolytic deamination of cytosine to give a miscoding uracil residue. This is also a frequent DNA lesion induced by oxidative damage, through exposure to agents such as ionizing radiation, or from endogenous sources that are implicated in the aetiology of degenerative diseases, ageing and cancer. The Ung and Smug1 enzymes excise uracil from DNA to effect repair in mammalian cells, and gene-targeted Ung(-/-) mice exhibit a moderate increase in genome-wide spontaneous mutagenesis. Here, we report that stable siRNA-mediated silencing of Smug1 in mouse embryo fibroblasts also generates a mutator phenotype. However, an additive 10-fold increase in spontaneous C:G to T:A transitions in cells deficient in both Smug1 and Ung demonstrates that these enzymes have distinct and nonredundant roles in suppressing C --> T mutability at non-CpG sites. Such cells are also hypersensitive to ionizing radiation, and reveal a role of Smug1 in the repair of lesions generated by oxidation of cytosine. PMID- 15902270 TI - Npl3 is an antagonist of mRNA 3' end formation by RNA polymerase II. AB - Proper 3' end formation is critical for the production of functional mRNAs. Termination by RNA polymerase II is linked to mRNA cleavage and polyadenylation, but it is less clear whether earlier stages of mRNA production also contribute to transcription termination. We performed a genetic screen to identify mutations that decreased transcriptional readthrough of a defective GAL10 poly(A) terminator. A partial deletion of the GAL10 downstream region leads to transcription through the downstream GAL7 promoter, resulting in the inability of cells to grow on galactose. Mutations in elongation factors Spt4 and Spt6 suppress the readthrough phenotype, presumably by decreasing the amount of polymerase transcribing through the downstream GAL7 promoter. Interestingly, mutations in the mRNA-binding protein Npl3 improve transcription termination. Both in vivo and in vitro experiments suggest that Npl3 can antagonize 3' end formation by competing for RNA binding with polyadenylation/termination factors. These results suggest that elongation rate and mRNA packaging can influence polyadenylation and termination. PMID- 15902271 TI - Calmodulin-dependent protein kinase IV regulates nuclear export of Cabin1 during T-cell activation. AB - Calcium signaling is critical for activation of T lymphocytes and has been proposed to be transduced through multiple calmodulin target proteins. Whereas the calcineurin-NFAT signaling module is critical for all mammalian T cells, the role of calmodulin-dependent kinase IV (CaMKIV) in mouse naive CD4+ T-cell activation remains enigmatic. We have applied lentivius-mediated RNA interference of CaMKIV to human T cells and found that knockdown of CaMKIV abrogates T-cell receptor-mediated transcription of the IL-2 gene. We demonstrate that CaMKIV directly phosphorylates Cabin1, a transcriptional corepressor for myocyte enhancer factor 2, creating a docking site for 14-3-3, which causes its nuclear export. CaMKIV-mediated nuclear export of Cabin1 is likely to account for a significant part of the requirement of CaMKIV during human T-cell activation. PMID- 15902272 TI - The MukF subunit of Escherichia coli condensin: architecture and functional relationship to kleisins. AB - The Escherichia coli MukB, MukE, and MukF proteins form a bacterial condensin (MukBEF) that contributes to chromosome management by compacting DNA. MukB is an ATPase and DNA-binding protein of the SMC superfamily; however, the structure and function of non-SMC components, such as MukF, have been less forthcoming. Here, we report the crystal structure of the N-terminal 287 amino acids of MukF at 2.9 A resolution. This region folds into a winged-helix domain and an extended coiled coil domain that self-associate to form a stable, doubly domain-swapped dimer. Protein dissection and affinity purification data demonstrate that the region of MukF C-terminal to this fragment binds to MukE and MukB. Our findings, together with sequence analyses, indicate that MukF is a kleisin subunit for E. coli condensin and suggest a means by which it may organize the MukBEF assembly. PMID- 15902273 TI - Homologous recombination is involved in transcription-coupled repair of UV damage in Saccharomyces cerevisiae. AB - To efficiently protect the integrity of genetic information, transcription is connected to nucleotide excision repair (NER), which allows preferential repair of the transcribed DNA strands (TS). As yet, the molecular basis of this connection remains elusive in eukaryotic cells. Here we show that, in haploids, the RAD26 gene is essential for the preferential repair of the TS during G1. However, in G2/M phase there is an additional RAD51-dependent process that enhances repair of TS. Importantly, the simultaneous deletion of both RAD26 and RAD51 led to complete abolishment of strand-specific repair during G2/M, indicating that these genes act through two independent but complementary subpathways. In diploids, however, RAD51 is involved in repair of the TS even in G1 phase, which unveils the implication of homologous recombination in the preferential repair of the TS. Importantly, the abolishment of NER, by abrogation of RAD1 or RAD14, completely stopped repair of UV damage even during G2/M phase. These results show the existence of functional cross-talk between transcription, homologous recombination and NER. PMID- 15902275 TI - No association between E- and L-selectin genes and SLE: soluble L-selectin levels do correlate with genotype and a subset in SLE. AB - Altered function of selectin glycoprotein adhesion molecules may modulate severity and organ-specific manifestations of autoimmune and inflammatory disease via changes in leukocyte trafficking. Serum concentrations of selectin molecules have been suggested as useful biomarkers in systemic lupus erythematosus (SLE). We identified increased levels of soluble L-selectin (sL-selectin), but not soluble E-selectin (sE-selectin) in 278 European-Caucasian lupus patients compared to 230 healthy siblings (P=0.002). sL-selectin levels were markedly elevated in patients with IgG antiphospholipid autoantibodies (P=0.002), suggesting that perhaps sL-selectin defines a subgroup of lupus with vasculopathy. sL-selectin level was also influenced by two L-selectin polymorphisms: 665C>T, F206L in the epidermal growth factor-like domain (P=0.015) and rs12938 in the 3'-untranslated region (P=0.06). Having shown increased sL selectin levels in lupus patients, we used genetics to investigate whether this was a secondary phenomena or the result of an underlying genetic mechanism. The inheritance of nine single-nucleotide polymorphisms (SNP) spanning the selectin locus was tested in 523 UK simplex SLE families. No association with SLE, or related phenotypes, was evident with any single SNP, or haplotype in family-based tests of association. Selectin polymorphisms are, therefore, unlikely to be independent factors in SLE susceptibility. PMID- 15902274 TI - The macro domain is an ADP-ribose binding module. AB - The ADP-ribosylation of proteins is an important post-translational modification that occurs in a variety of biological processes, including DNA repair, transcription, chromatin biology and long-term memory formation. Yet no protein modules are known that specifically recognize the ADP-ribose nucleotide. We provide biochemical and structural evidence that macro domains are high-affinity ADP-ribose binding modules. Our structural analysis reveals a conserved ligand binding pocket among the macro domain fold. Consistently, distinct human macro domains retain their ability to bind ADP-ribose. In addition, some macro domain proteins also recognize poly-ADP-ribose as a ligand. Our data suggest an important role for proteins containing macro domains in the biology of ADP ribose. PMID- 15902276 TI - Protein toxins: intracellular trafficking for targeted therapy. AB - The immunotoxin approach is based on the use of tumor-targeting ligands or antibodies that are linked to the catalytic (toxic) moieties of bacterial or plant protein toxins. In this review, we first discuss the current state of clinical development of immunotoxin approaches describing the results obtained with the two toxins most frequently used: diphtheria and Pseudomonas toxin derived proteins. In the second part of the review, a novel concept will be presented in which the roles are inverted: nontoxic receptor-binding toxin moieties are used for the targeting of therapeutic and diagnostic compounds to cancer or immune cells. The cell biological basis of these novel types of toxin based therapeutics will be discussed, and we will summarize ongoing preclinical and clinical testing. PMID- 15902277 TI - Evaluation of a synthetic CArG promoter for nitric oxide synthase gene therapy of cancer. AB - Nitric oxide synthase gene therapy has been shown to be effective at inducing apoptosis in experimental tumours and sensitizing them to radiotherapy. We have also shown that expression of inducible nitric oxide synthase (iNOS) can be effectively restricted to the tumour volume by the use of the radiation inducible promoter (WAF1) to drive the transgene in clinically relevant protocols. A synthetic construct (pE9), incorporating nine radiosensitive CArG elements from the Egr1 promoter, has recently been developed for cancer gene therapy. We have now investigated basal gene expression of transgenes driven by this promoter to assess its suitability for use in iNOS gene therapy protocols in vivo. Transfection of human microvascular endothelial cells (HMEC-1) with pE9iNOS, using a cationic lipid vector, resulted in progressively increasing (<5-fold) levels of iNOS protein expression up to 8 h after transfection. Transfection of an ex vivo rat artery preparation with pE9iNOS caused 83% inhibition of response to the vasoconstrictor phenylephrine (PE). CMViNOS transfection also reduced response to PE, but by only 52%. A single injection of 25 microg of pE9iNOS DNA in a lipid vector into the centre of a murine sarcoma (RIF1) induced iNOS protein expression by four-fold and increased nitrite concentration eight-fold. This caused a 7-day delay in tumour growth and was more effective than the constitutive CMV-driven construct. Our data suggest that generation of NO*, as a result of iNOS overexpression, is capable of further activating the E9 promoter, through a positive feedback loop, yielding stronger and sustained levels of NO*. This pE9iNOS combination may, therefore, be particularly useful in an anticancer gene therapy strategy as its antitumour effect in vivo was clearly superior to that of the strong constitutive promoter, CMV. PMID- 15902278 TI - Neurovascular bundle block as predictor of postoperative erectile function after radical prostatectomy. AB - We assessed audiovisually induced erections after nerve block of the neurovascular bundle during prostate biopsy. We evaluated neurovascular bundle nerve block to mimic non-nerve-sparing radical prostatectomy in an experimental setup. Patients undergoing a transrectal ultrasound-guided prostate biopsy were randomized to bilateral injection of 5 ml ropivacaine hydrochloride 0.75% or NaCl 0.9% into the neurovascular bundle. The patients completed the International Index of Erectile Function 5-item questionnaire (IIEF-5) questionnaire, and a detailed patient history was obtained. A routine prostate biopsy was performed. Thereafter, patients were exposed to 60 min of audiovisual stimulation. Erections were recorded using a Rigiscan-Plus device. A total of 11 patients were randomized. Five patients received NaCl (group 1) and six patients ropivacaine (group 2). Patient characteristics were comparable in terms of age (group 1: 59.8 y; group 2: 61.8 y), mean PSA (4.1 vs 4.7 ng/ml), mean IIEF-5 score (20.5 vs 22) and risk factors for erectile dysfunction, respectively. Patients of group 1 showed significantly stronger and longer erections after audiovisual stimulation than patients in group 2. Patients with bilateral infiltration of saline solution to the neurovascular bundle showed significantly stronger erections than patients receiving local anesthesia of the neurovascular bundle. Thus, this experiment might serve as a model to assess postoperative erectile function after a unilateral nerve-sparing radical prostatectomy. PMID- 15902279 TI - Obesity, the metabolic syndrome, and sexual dysfunction. AB - Sexual problems in both sexes appear to be widespread in society, influenced by both health-related and psychosocial factors, and are associated with impaired quality of life. Epidemiological studies suggest that modifiable health behaviors, including physical activity and leanness, are associated with a reduced risk for erectile dysfunction (ED) among men. Data from other surveys also indicate a higher prevalence of impotence in obese men. Obesity may be a risk factor for sexual dysfunction in both sexes; the data for the metabolic syndrome are very preliminary and need to be confirmed in larger epidemiologic studies. The high prevalence of ED in patients with cardiovascular risk factors suggests that abnormalities of the vasodilator system of penile arteries play an important role in the pathophysiology of ED. We have shown that one-third of obese men with ED can regain their sexual activity after 2 y of adopting health behaviors, mainly regular exercise and reducing weight. Western societies actually spend a huge part of their health care costs on chronic disease treatment and interventions for risk factors. The adoption of healthy lifestyles can reduce the prevalence of obesity and the metabolic syndrome, and hopefully the burden of sexual dysfunction. PMID- 15902281 TI - Genomic gains and losses influence expression levels of genes located within the affected regions: a study on acute myeloid leukemias with trisomy 8, 11, or 13, monosomy 7, or deletion 5q. AB - We performed microarray analyses in AML with trisomies 8 (n=12), 11 (n=7), 13 (n=7), monosomy 7 (n=9), and deletion 5q (n=7) as sole changes to investigate whether genomic gains and losses translate into altered expression levels of genes located in the affected chromosomal regions. Controls were 104 AML with normal karyotype. In subgroups with trisomy, the median expression of genes located on gained chromosomes was higher, while in AML with monosomy 7 and deletion 5q the median expression of genes located in deleted regions was lower. The 50 most differentially expressed genes, as compared to all other subtypes, were equally distributed over the genome in AML subgroups with trisomies. In contrast, 30 and 86% of the most differentially expressed genes characteristic for AML with 5q deletion and monosomy 7 are located on chromosomes 5 or 7. In conclusion, gain of whole chromosomes leads to overexpression of genes located on the respective chromosomes. Losses of larger regions of the genome translate into lower expression of the majority of genes represented by only one allele. The reduced expression of these genes is the most characteristic difference in gene expression profiles between AML with monosomy 7 and AML with deletion 5q, respectively, and other AML subtypes. Therefore, these data provide evidence that gene dosage effects gene expression in AML with unbalanced karyotype abnormalities. Losses of specific regions of the genome determine the gene expression profile more strongly than the gain of whole chromosomes. PMID- 15902282 TI - Expression and methylation status of the FHIT gene in acute myeloid leukemia and myelodysplastic syndrome. AB - To clarify the role of fragile histidine triad (FHIT) in hematological malignancies, we examined the methylation status and the expression level of the FHIT gene in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) cells in comparison with the methylation of the p15(INK4B) gene. The FHIT methylation was found in 13 of 94 (13.8%) AML and 22 of 40 (55.0%) MDS cases, but not in normal mononuclear cells (MNCs). Both the frequency and density of methylation increased in the advanced-stages MDS and the relapsed AML cases. Although FHIT and p15(INK4B) methylations were not correlated in MDS and AML, increased FHIT methylation at the relapse in AML was associated with p15(INK4B) methylation. The median expression level in AML was significantly higher than in normal MNCs, although the median expression level in those with methylation was significantly lower than in those without methylation. Furthermore, the methylation level at relapse was significantly higher than at diagnosis in AML. These results suggested that FHIT methylation was accumulated through the disease progression of MDS and AML, and the role of the FHIT gene as a tumor suppressor seemed different in AML and MDS. PMID- 15902283 TI - Is it chronic idiopathic myelofibrosis, myelofibrosis with myeloid metaplasia, chronic megakaryocytic-granulocytic myelosis, or chronic megakaryocytic leukemia? Further thoughts on the nosology of the clonal myeloid disorders. PMID- 15902284 TI - Mutations in the receptor tyrosine kinase pathway are associated with clinical outcome in patients with acute myeloblastic leukemia harboring t(8;21)(q22;q22). AB - AML1-MTG8 generated by t(8;21) contributes to leukemic transformation, but additional events are required for full leukemogenesis. We examined whether mutations in the receptor tyrosine kinase (RTK) pathway could be the genetic events that cause acute myeloblastic leukemia (AML) harboring t(8;21). Mutations in the second tyrosine kinase domain, juxtamembrane (JM) domain and exon 8 of the C-KIT gene were observed in 10, one and three of 37 AML patients with t(8;21), respectively. Three patients showed an internal tandem duplication in the JM domain of the FLT3 gene. One patient had a mutation in the K-Ras gene at codon 12. As the occurrence of these mutations was mutually exclusive, a total of 18 (49%) patients showed mutations in the RTK pathway. These results suggest that activating mutations in the RTK pathway play a role in part as an additional event leading to the development of t(8;21) AML. The 6-year cumulative incidence of relapse in patients with RTK pathway mutations was 79.8%, compared with 13.5% in patients lacking such mutations (P=0.0029). Furthermore, the 6-year relapse free survival in patients with mutations was 18% compared to 60% in those without mutations (P=0.0340), indicating that RTK mutations are associated with the clinical outcome in t(8;21) AML. PMID- 15902285 TI - A limited role for TP53 mutation in the transformation of follicular lymphoma to diffuse large B-cell lymphoma. AB - The role of TP53 mutation in transformation of follicular lymphoma (FL) to diffuse large B-cell lymphoma (t-FL) was examined in a panel of 91 lymph node biopsies derived from 29 patients pre- and post-transformation. The entire TP53 coding sequence was screened and immunocytochemistry performed to determine expression of p53 and its key regulator MDM2. A total of 10 mutations were detected in eight patients (28%), although none were present at FL diagnosis. Mutations were not detected solely at the time of transformation; in three patients, mutated TP53 arose in at least one antecedent FL sample (6 months, 2.5 years and 4 years prior to transformation). Loss of heterozygosity at the TP53 locus occurred in 2/20 informative patients (only in t-FL samples). p53 staining was positive in 82% (9/11) of available biopsies with a missense mutation, and negative in 71% (45/63) with wtTP53. MDM2 expression was significantly higher in t-FL samples (mean 72% positive; 95% confidence interval (95% CI) 68-76%) than FL (mean 58% positive; 95% CI 54-62%) (P<0.001) but did not correlate with TP53 status. TP53 mutation has only a limited role in the transformation of FL, exerting a heterogeneous influence upon phenotypic change. In contrast, dysregulation of MDM2 is frequent and may provide a more rational therapeutic target.. PMID- 15902286 TI - Higher-grade transformation of follicular lymphoma -- a continuous enigma. PMID- 15902287 TI - What have we learnt from mouse models of NPM-ALK-induced lymphomagenesis? AB - The nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) is generated as a t(2;5) chromosomal breakpoint product, typically in CD30(+) anaplastic large cell lymphomas. Activation of the NPM-ALK tyrosine kinase by NPM dimerisation causes autophosphorylation at multiple tyrosine residues and the consequent recruitment of a 'signalosome' that couples the fusion protein to pathways regulating mitogenesis and apoptosis. This review focuses on recent advances in our understanding of the transforming signals induced by this fusion protein in mouse models. PMID- 15902288 TI - Bone marrow as a home of heterogenous populations of nonhematopoietic stem cells. AB - Evidence is presented that bone marrow (BM) in addition to CD45(positive) hematopoietic stem cells contains a rare population of heterogenous CD45(negative) nonhematopoietic tissue committed stem cells (TCSC). These nonhematopoietic TCSC (i) are enriched in population of CXCR4(+) CD34(+) AC133(+) lin(-) CD45(-) and CXCR4(+) Sca-1(+) lin(-) CD45(-) in humans and mice, respectively, (ii) display several markers of pluripotent stem cells (PSC) and (iii) as we envision are deposited in BM early in development. Thus, since BM contains versatile nonhematopoietic stem cells, previous studies on plasticity trans-dedifferentiation of BM-derived hematopoietic stem cells (HSC) that did not include proper controls to exclude this possibility could lead to wrong interpretations. Therefore, in this spotlight review we present this alternative explanation of 'plasticity' of BM-derived stem cells based on the assumption that BM stem cells are heterogenous. We also discuss a potential relationship of TCSC/PSC identified by us with other BM-derived CD45(negative) nonhematopoietic stem cells that were recently identified by other investigators (eg MSC, MAPC, USSC and MIAMI cells). Finally, we discuss perspectives and pitfalls in potential application of these cells in regenerative medicine. PMID- 15902289 TI - Mesenchymal stem cells can be easily isolated from bone marrow of patients with various haematological malignancies but the surface antigens expression may be changed after prolonged ex vivo culture. PMID- 15902291 TI - Treatment of relapsed acute myelogeneous leukaemia with MLL/AF 6 fusion after stem cell transplantation by intensive reinduction followed by adoptive immunotherapy. PMID- 15902292 TI - Further evidence that germline CEBPA mutations cause dominant inheritance of acute myeloid leukaemia. PMID- 15902293 TI - Occurrence of de novo ABL kinase domain mutations in primary bone marrow cells after BCR-ABL gene transfer and Imatinib mesylate selection. PMID- 15902294 TI - Mcl-1 is overexpressed in multiple myeloma and associated with relapse and shorter survival. AB - We and others have shown that Mcl-1 was essential for the survival of human myeloma cells in vitro. Furthermore, this antiapoptotic protein is upregulated by interleukin-6, which plays a critical role in multiple myeloma (MM). For these reasons, we have evaluated the expression of Mcl-1 in vivo in normal, reactive and malignant plasma cells (PC), that is, myeloma cells from 51 patients with MM and 21 human myeloma cell lines (HMCL) using flow cytometry. We show that Mcl-1 is overexpressed in MM in comparison with normal bone marrow PC. In total, 52% of patients with MM at diagnosis (P=0.017) and 81% at relapse (P=0.014 for comparison with diagnosis) overexpress Mcl-1. Of note, only HMCL but not reactive plasmacytoses have abnormal Mcl-1 expression, although both PC expansions share similar high proliferation rates. Of interest, Bcl-2 as opposed to Mcl-1, does not discriminate malignant from normal PC. Finally, the level of Mcl-1 expression is related to disease severity, the highest values at diagnosis being associated with the shortest event-free survival (P=0.002). In conclusion, Mcl-1, which has been shown to be essential for the survival of human myeloma cells in vitro, is overexpressed in vivo in MM in relation with relapse and shorter survival. Mcl-1 represents a potential therapeutical target in MM. PMID- 15902295 TI - The seventh international childhood acute lymphoblastic leukemia workshop report: Palermo, Italy, January 29--30, 2005. AB - Between 1995 and 2004, six International Childhood Acute Lymphoblastic Leukemia (ALL) Workshop have been held, and the completion of several collaborative projects has established the clinical relevance and treatment options for several specific genetic subtypes of ALL. This meeting report summarizes the data presented in the seventh meeting and the discussion. PMID- 15902296 TI - Protein expression analysis of chromosome 12 candidate genes in chronic lymphocytic leukemia (CLL). AB - The pathogenic role of trisomy 12 in chronic lymphocytic leukemia (CLL) remains unresolved, but recently an upregulated RNA expression level has been observed for chromosome 12 candidate genes. In the current study, the protein expression of chromosome 12 candidate genes was characterized by comparing CLL cases with (n=58) or without (n=16) trisomy 12, CD19+-B-cells and cell lines (JVM-2, EHEB, JURKAT). Immunoblotting was performed to quantify the levels of AID, APAF-1, ARF3, CCND2, CDK2, CKD4, GLI, MDM-2, p27, Smac/DIABLO and STAT6 (signal transducer and activator of transcription 6). The cell lines showed distinct expression patterns for CCND2, MDM-2, p27, Smac/DIABLO and STAT6, and displayed higher levels of CDK2 and CDK4 than the CLL cases. JURKAT and the CLL cases expressed uniformly high levels of p27, but low levels of CCND2. AID expression in the CLL cases was weak with slight variations regardless of the subgroup affiliation. The expression of the investigated proteins was independent of the trisomy 12 status as well as of the VH mutation status. The comparison of CD19+-B cells with CLL revealed higher protein levels in CLL for CDK4, p27, Smac/DIABLO and STAT6. Further studies including protein expression experiments in genetic high-risk subgroups of CLL have to elucidate whether these proteins qualify as candidates for targeted CLL therapies. PMID- 15902297 TI - The histone deacetylase inhibitor valproic acid alters sensitivity towards all trans retinoic acid in acute myeloblastic leukemia cells. AB - Acute myeloblastic leukemia (AML) may be classified in a number of ways. Using the French American British classification, the M3 form of the disease or acute promyelocytic leukemia (APL) has been found to be sensitive in vitro and in vivo to the retinoid all trans retinoic acid (ATRA). The mechanism for this is by restoration of normal gene expression through the release of histone deacetylase complexes (HDACs). In contrast to APL, other forms of AML are either nonresponsive or show blunted responses to ATRA. We evaluated if the inhibitor of HDAC activity, valproic acid (VPA), could mimic or enhance retinoid sensitivity in the AML cell line, OCI/AML-2, and clinical samples derived from patients with AML. An Affymetrix GeneChip experiment demonstrated that VPA modulated the expression of numerous genes in OCI/AML-2 cells that were not affected by ATRA including p21, a retinoid responsive gene in APL. VPA induced p21 expression in OCI/AML-2 cells and the majority of the AML samples tested; this was associated with cell cycle arrest and apoptosis not seen with ATRA alone. The addition of ATRA to VPA accentuated many of these responses, supporting the potential beneficial combination of these drugs in the treatment of AML. PMID- 15902298 TI - Synergistic activity of imatinib and 17-AAG in imatinib-resistant CML cells overexpressing BCR-ABL--Inhibition of P-glycoprotein function by 17-AAG. AB - Overexpression of BCR-ABL and P-glycoprotein (Pgp) are two of the known mechanisms of imatinib resistance. As combination therapy may allow to overcome drug resistance, we investigated the effect of combination treatment with imatinib and 17-allylamino-17-demethoxygeldanamycin (17-AAG), a heat-shock protein 90 (Hsp90) inhibitor, on different imatinib-sensitive and imatinib resistant CML cell lines. In imatinib-sensitive cells, combination index (CI) values obtained using the method of Chou and Talalay indicated additive (CI=1) or marginally antagonistic (CI>1) effects following simultaneous treatment with imatinib and 17-AAG. In imatinib-resistant cells both drugs acted synergistically (CI<1). In primary chronic-phase CML cells additive or synergistic effects of the combination of imatinib plus 17-AAG were discernible. Annexin V/propidium iodide staining showed that the activity of imatinib plus 17-AAG is mediated by apoptosis. Combination treatment with imatinib plus 17-AAG was more effective in reducing the BCR-ABL protein level than 17-AAG alone. Monotherapy with 17-AAG decreased P-glycoprotein activity, which may increase intracellular imatinib levels and contribute to the sensitization of CML cells to imatinib. The results suggest that combination of imatinib and 17-AAG may be useful to overcome imatinib resistance in a clinical setting. PMID- 15902299 TI - Desferrioxamine induces leukemic cell differentiation potentially by hypoxia inducible factor-1 alpha that augments transcriptional activity of CCAAT/enhancer binding protein-alpha. AB - We reported recently that cobalt chloride-simulated hypoxia and mild hypoxia modified the differentiation of human acute myeloid leukemic (AML) cells, probably acting via a hypoxia-inducible factor-1 alpha (HIF-1 alpha)-dependent mechanism. In this study, we investigated the effect of desferrioxamine (DFO), an iron chelator with 'hypoxia-mimetic' activity, on the differentiation of AML cells. The results showed that DFO at nontoxic concentrations induced the differentiation of AML cell lines NB4 and U937, as assessed by morphological criteria and differentiation-associated antigens. DFO-induced differentiation parallel to the rapid accumulation of HIF-1 alpha protein in these two cell lines. Of importance, the transient transfection of HIF-1 alpha cDNA induced U937 cells to develop the differentiation-related alterations such as growth arrest and increased CD11b expression. Furthermore, the inducible expression of chromosome translocation t(8;21)-generated leukemogenic AML1-ETO fusion gene attenuated DFO-induced differentiation of U937 cells with the decrease of CCAAT/enhancer-binding protein alpha (C/EBP alpha), a critical factor for granulocytic differentiation. Using immunoprecipitation and luciferase reporter assay, HIF-1 alpha was also shown to interact physically with and to increase the transcriptional activity of C/EBP alpha. Taken together, these results provided novel evidence for a role of HIF-1 alpha in AML cell differentiation, and suggested that C/EBP alpha might be a downstream effector for HIF-1 alpha mediated differentiation. PMID- 15902300 TI - Significance of chemokine receptor expression in aggressive NK cell leukemia. AB - Natural killer (NK) cell-type lymphoproliferative diseases of granular lymphocytes can be subdivided into aggressive NK cell leukemia (ANKL) and chronic NK cell lymphocytosis (CNKL). One reason for the poor outcome in ANKL is leukemic infiltration into multiple organs. The mechanisms of cell trafficking associated with the chemokine system have been investigated in NK cells. To clarify the mechanism of systemic migration of leukemic NK cells, we enrolled nine ANKL and six CNKL cases, and analyzed the expression profiles and functions of chemokine receptors by flowcytometry and chemotaxis assay. CXCR1 was detected on NK cells in all groups, and CCR5 was positive in all ANKL cells. Proliferating NK cells were simultaneously positive for CXCR1 and CCR5 in all ANKL patients examined, and NK cells with this phenotype did not expand in CNKL patients or healthy donors. ANKL cells showed enhanced chemotaxis toward the ligands of these receptors. These results indicated that the chemokine system might play an important role in the pathophysiology of ANKL and that chemokine receptor profiling might be a novel tool for discriminating ANKL cells from benign NK cells. PMID- 15902301 TI - Mice with a severe deficiency in protein C display prothrombotic and proinflammatory phenotypes and compromised maternal reproductive capabilities. AB - Anticoagulant protein C (PC) is important not only for maintenance of normal hemostasis, but also for regulating the host immune response during inflammation. Because mice with a designed total genetic deficiency in PC (PC-/- mice) die soon after birth, attempts to dissect PC function in various coagulation/inflammation based pathologies through use of mice with less than 50% of normal PC levels have not been successful to date. In the current investigation, we have used a novel transgenic strategy to generate different mouse models expressing 1-18% of normal PC levels. In contrast to PC-/- mice, mice with only partial PC deficiency survived beyond birth and also developed thrombosis and inflammation. The onset and severity of these phenotypes vary significantly and are strongly dependent on plasma PC levels. Our findings additionally provide the first evidence that maternal PC is vital for sustaining pregnancy beyond 7.5 days postcoitum, likely by regulating the balance of coagulation and inflammation during trophoblast invasion. These low PC-expressing transgenic mouse lines provide novel animal models that can be used to elucidate the importance of PC in maintenance of the organism and in disease. PMID- 15902302 TI - Enhanced passive Ca2+ reabsorption and reduced Mg2+ channel abundance explains thiazide-induced hypocalciuria and hypomagnesemia. AB - Thiazide diuretics enhance renal Na+ excretion by blocking the Na+-Cl- cotransporter (NCC), and mutations in NCC result in Gitelman syndrome. The mechanisms underlying the accompanying hypocalciuria and hypomagnesemia remain debated. Here, we show that enhanced passive Ca2+ transport in the proximal tubule rather than active Ca2+ transport in distal convolution explains thiazide induced hypocalciuria. First, micropuncture experiments in mice demonstrated increased reabsorption of Na+ and Ca2+ in the proximal tubule during chronic hydrochlorothiazide (HCTZ) treatment, whereas Ca2+ reabsorption in distal convolution appeared unaffected. Second, HCTZ administration still induced hypocalciuria in transient receptor potential channel subfamily V, member 5 knockout (Trpv5-knockout) mice, in which active distal Ca2+ reabsorption is abolished due to inactivation of the epithelial Ca2+ channel Trpv5. Third, HCTZ upregulated the Na+/H+ exchanger, responsible for the majority of Na+ and, consequently, Ca2+ reabsorption in the proximal tubule, while the expression of proteins involved in active Ca2+ transport was unaltered. Fourth, experiments addressing the time-dependent effect of a single dose of HCTZ showed that the development of hypocalciuria parallels a compensatory increase in Na+ reabsorption secondary to an initial natriuresis. Hypomagnesemia developed during chronic HCTZ administration and in NCC-knockout mice, an animal model of Gitelman syndrome, accompanied by downregulation of the epithelial Mg2+ channel transient receptor potential channel subfamily M, member 6 (Trpm6). Thus, Trpm6 downregulation may represent a general mechanism involved in the pathogenesis of hypomagnesemia accompanying NCC inhibition or inactivation. PMID- 15902303 TI - Unmutated and mutated chronic lymphocytic leukemias derive from self-reactive B cell precursors despite expressing different antibody reactivity. AB - B cell chronic lymphocytic leukemia (CLL) is a disease of expanding monoclonal B cells whose B cell receptor (BCR) mutational status defines 2 subgroups; patients with mutated BCRs have a more favorable prognosis than those with unmutated BCRs. CLL B cells express a restricted BCR repertoire including antibodies with quasi identical complementarity-determining region 3 (CDR3), which suggests specific antigen recognition. The antigens recognized by CLL antibodies may include autoantigens since about half of CLL B cells produce autoreactive antibodies. However, the distribution of autoreactive antibodies between Ig heavy-chain variable-unmutated (IgV-unmutated) CLL (UM-CLL) and IgV-mutated CLL (M-CLL) is unknown. To determine the role of antibody reactivity and the impact of somatic hypermutation (SHM) on CLL antibody specificity, we cloned and expressed in vitro recombinant antibodies from M- and UM-CLL B cells and tested their reactivity by ELISA. We found that UM-CLL B cells expressed highly polyreactive antibodies whereas most M-CLL B cells did not. When mutated nonautoreactive CLL antibody sequences were reverted in vitro to their germline counterparts, they encoded polyreactive and autoreactive antibodies. We concluded that both UM-CLLs and M CLLs originate from self-reactive B cell precursors and that SHM plays an important role in the development of the disease by altering original BCR autoreactivity. PMID- 15902304 TI - Ferroportin1 is required for normal iron cycling in zebrafish. AB - Missense mutations in ferroportin1 (fpn1), an intestinal and macrophage iron exporter, have been identified between transmembrane helices 3 and 4 in the zebrafish anemia mutant weissherbst (weh(Tp85c-/-)) and in patients with type 4 hemochromatosis. To explore the effects of fpn1 mutation on blood development and iron homeostasis in the adult zebrafish, weh(Tp85c-/-) zebrafish were rescued by injection with iron dextran and studied in comparison with injected and uninjected WT zebrafish and heterozygotes. Although iron deposition was observed in all iron-injected fish, only weh(Tp85c-/-) zebrafish exhibited iron accumulation in the intestinal epithelium compatible with a block in iron export. Iron injections initially reversed the anemia. However, 8 months after iron injections were discontinued, weh(Tp85c-/-) zebrafish developed hypochromic anemia and impaired erythroid maturation despite the persistence of iron-loaded macrophages and elevated hepatic nonheme iron stores. Quantitative real-time RT PCR revealed a significant decrease in mean hepatic transcript levels of the secreted iron-regulator hepcidin and increased intestinal expression of fpn1 in anemic weh(Tp85c-/-) adults. Injection of iron dextran into WT or mutant zebrafish embryos, however, resulted in significant increases in hepcidin expression 18 hours after injection, demonstrating that hepcidin expression in zebrafish is iron responsive and independent of fpn1's function as an iron exporter. PMID- 15902305 TI - Morphogenesis of the right ventricle requires myocardial expression of Gata4. AB - Mutations in developmental regulatory genes have been found to be responsible for some cases of congenital heart defects. One such regulatory gene is Gata4, a zinc finger transcription factor. In order to circumvent the early embryonic lethality of Gata4-null embryos and to investigate the role of myocardial Gata4 expression in cardiac development, we used Cre/loxP technology to conditionally delete Gata4 in the myocardium of mice at an early and a late time point in cardiac morphogenesis. Early deletion of Gata4 by Nkx2-5Cre resulted in hearts with striking myocardial thinning, absence of mesenchymal cells within the endocardial cushions, and selective hypoplasia of the RV. RV hypoplasia was associated with downregulation of Hand2, a transcription factor previously shown to regulate formation of the RV. Cardiomyocyte proliferation was reduced, with a greater degree of reduction in the RV than in the LV. Late deletion of Gata4 by Cre recombinase driven by the alpha myosin heavy chain promoter did not selectively affect RV development or generation of endocardial cushion mesenchyme but did result in marked myocardial thinning with decreased cardiomyocyte proliferation, as well as double-outlet RV. Our results demonstrate a general role of myocardial Gata4 in regulating cardiomyocyte proliferation and a specific, stage-dependent role in regulating the morphogenesis of the RV and the atrioventricular canal. PMID- 15902306 TI - FGF-21 as a novel metabolic regulator. AB - Diabetes mellitus is a major health concern, affecting more than 5% of the population. Here we describe a potential novel therapeutic agent for this disease, FGF-21, which was discovered to be a potent regulator of glucose uptake in mouse 3T3-L1 and primary human adipocytes. FGF-21-transgenic mice were viable and resistant to diet-induced obesity. Therapeutic administration of FGF-21 reduced plasma glucose and triglycerides to near normal levels in both ob/ob and db/db mice. These effects persisted for at least 24 hours following the cessation of FGF-21 administration. Importantly, FGF-21 did not induce mitogenicity, hypoglycemia, or weight gain at any dose tested in diabetic or healthy animals or when overexpressed in transgenic mice. Thus, we conclude that FGF-21, which we have identified as a novel metabolic factor, exhibits the therapeutic characteristics necessary for an effective treatment of diabetes. PMID- 15902307 TI - Modulation of bone morphogenetic protein signaling inhibits the onset and progression of ankylosing enthesitis. AB - Joint ankylosis is a major cause of disability in the human spondyloarthropathies. Here we report that this process partially recapitulates embryonic endochondral bone formation in a spontaneous model of arthritis in DBA/1 mice. Bone morphogenetic protein (BMP) signaling appears to be a key molecular pathway involved in this pathological cascade. Systemic gene transfer of noggin, a BMP antagonist, is effective both as a preventive and a therapeutic strategy in the mouse model, mechanistically interfering with enthesial progenitor cell proliferation in early stages of the disease process. Immunohistochemical staining for phosphorylated smad1/5 in enthesial biopsies of patients with spondyloarthropathy reveals active BMP signaling in similar target cells. Our data suggest that BMP signaling is an attractive therapeutic target for interfering with structural changes in spondyloarthropathy either as an alternative or complementary approach to current antiinflammatory treatments. PMID- 15902308 TI - Integrin alpha4beta1-VCAM-1-mediated adhesion between endothelial and mural cells is required for blood vessel maturation. AB - Neovascularization depends on vascular cell proliferation and on the stabilization of vessels by association of vascular smooth muscle-like pericytes with ECs. Here we show that integrin alpha4beta1 (VLA-4) and VCAM-1 promote close intercellular adhesion between ECs and pericytes and that this interaction is required for blood vessel formation. Integrin alpha4beta1 is expressed by proliferating but not quiescent ECs, while its ligand VCAM-1 is expressed by proliferating but not quiescent mural cells. Antagonists of this integrin-ligand pair block the adhesion of mural cells to proliferating endothelia in vitro and in vivo, thereby inducing apoptosis of ECs and pericytes and inhibiting neovascularization. These studies indicate that integrin alpha4beta1 and VCAM-1 facilitate a critical cell-cell adhesion event required for survival of endothelial and mural cells during vascularization. PMID- 15902309 TI - Genome-wide expression analysis of therapy-resistant tumors reveals SPARC as a novel target for cancer therapy. AB - Overcoming resistance to chemotherapy and radiation therapy has been a difficult but important goal in the effort to cure cancer. We used gene-expression microarrays to identify differentially expressed genes involved in colorectal cancer resistance to chemotherapy and identified secreted protein, acidic and rich in cysteine (osteonectin) (SPARC) as a putative resistance-reversal gene by demonstrating low SPARC expression in refractory human MIP101 colon cancer cells. We were able to achieve restoration of their radiosensitivity and sensitivity to 5-fluorouracil and irinotecan by reexpression of SPARC in tumor xenografts. Moreover, treatment of mice with SPARC conferred increased sensitivity to chemotherapy and led to significant regression of xenografted tumors. The results show that modulation of SPARC expression affects colorectal cancer sensitivity to radiation and chemotherapy. SPARC-based gene or protein therapy may ameliorate the emergence of resistant clones and eradicate existing refractory clones and offers a novel approach to treating cancer. PMID- 15902310 TI - Bacterial programmed cell death of cerebral endothelial cells involves dual death pathways. AB - Major barriers separating the blood from tissue compartments in the body are composed of endothelial cells. Interaction of bacteria with such barriers defines the course of invasive infections, and meningitis has served as a model system to study endothelial cell injury. Here we report the impressive ability of Streptococcus pneumoniae, clinically one of the most important pathogens, to induce 2 morphologically distinct forms of programmed cell death (PCD) in brain derived endothelial cells. Pneumococci and the major cytotoxins H2O2 and pneumolysin induce apoptosis-like PCD independent of TLR2 and TLR4. On the other hand, pneumococcal cell wall, a major proinflammatory component, causes caspase driven classical apoptosis that is mediated through TLR2. These findings broaden the scope of bacterial-induced PCD, link these effects to innate immune TLRs, and provide insight into the acute and persistent phases of damage during meningitis. PMID- 15902311 TI - Role of C5 in the development of airway inflammation, airway hyperresponsiveness, and ongoing airway response. AB - The role of complement component C5 in asthma remains controversial. Here we examined the contribution of C5 at 3 critical checkpoints during the course of disease. Using an mAb specific for C5, we were able to evaluate the contribution of C5 during (a) the initiation of airway inflammation, (b) the maintenance of airway hyperresponsiveness (AHR), and (c) sustainment of an ongoing airway response to allergen provocation. Our results indicate that C5 is probably activated intrapulmonarily after infections or exposures to allergen and C5 inhibition has profound effects at all 3 critical checkpoints. In contrast to an earlier report, C5-deficient mice with established airway inflammation did not have elevated AHR to nonspecific stimuli. In the presence of airway inflammation, C5a serves as a direct link between the innate immune system and the development of AHR by engaging directly with its receptors expressed in airways. Through their powerful chemotactic and cell activation properties, both C5a and C5b-9 regulate the downstream inflammatory cascade, which results in a massive migration of inflammatory cells into the bronchial airway lumen and triggers the release of multiple harmful inflammatory mediators. This study suggests that targeting C5 is a potential clinical approach for treating patients with asthma. PMID- 15902313 TI - Loss of GABAergic Interneurons in Seizure-induced Epileptogenesis. PMID- 15902314 TI - H-channels as a Therapeutic Target in Epilepsy. PMID- 15902312 TI - Antigen-specific CD4+ T cells drive airway smooth muscle remodeling in experimental asthma. AB - Airway smooth muscle (ASM) growth contributes to the mechanism of airway hyperresponsiveness in asthma. Here we demonstrate that CD4+ T cells, central to chronic airway inflammation, drive ASM remodeling in experimental asthma. Adoptive transfer of CD4+ T cells from sensitized rats induced an increase in proliferation and inhibition of apoptosis of airway myocytes in naive recipients upon repeated antigen challenge, which resulted in an increase in ASM mass. Genetically modified CD4+ T cells expressing enhanced GFP (EGFP) were localized by confocal microscopy in juxtaposition to ASM cells, which suggests that CD4+ T cells may modulate ASM cell function through direct cell-cell interaction in vivo. Coculture of antigen-stimulated CD4+ T cells with cell cycle-arrested ASM cells induced myocyte proliferation, dependent on T cell activation and direct T cell-myocyte contact. Reciprocally, direct cell contact prevented postactivation T cell apoptosis, which suggests receptor-mediated T cell-myocyte crosstalk. Overall, our data demonstrate that activated CD4+ T cells drive ASM remodeling in experimental asthma and suggest that a direct cell-cell interaction participates in CD4+ T cell regulation of myocyte turnover and induction of remodeling. PMID- 15902315 TI - Long-term Outcomes of Temporal Lobe Epilepsy Surgery. PMID- 15902316 TI - A Potential Role for alpha-Methyl-l-tryptophan PET in Seizure Localization in Patients with Intractable Epilepsy. PMID- 15902317 TI - New Requirements for Registering Clinical Trials. PMID- 15902318 TI - Effects of Developmental Stage and Sex on Lower Extremity Kinematics and Vertical Ground Reaction Forces During Landing. AB - Context: The presence or absence of biomechanical differences between the sexes before puberty may provide clues about the onset of adult landing pattern differences, which may help to explain the greater number of anterior cruciate ligament injuries in females than in males and provide the basis for interventions to reduce those injuries.Objective: To identify developmental sex related and biomechanical differences during vertical jump landings.Design: A 2 x 2 developmental stage (prepubescent or postpubescent) x sex (male or female) between-subjects design.Setting: Controlled laboratory setting.Patients or Other Participants: Thirty prepubescent subjects (15 boys, age = 9.63 +/- 0.95 years; 15 girls, age = 9.19 +/- 1.00 years) and 28 postpubescent subjects (14 men, age = 23.57 +/- 3.23 years; 14 women, age = 24.22 +/- 2.27 years).Intervention: Subjects performed a vertical jump to a target set at 50% of their maximum vertical jump height ability.Main Outcome Measure(s): Hip and knee kinematics of the dominant lower extremity and vertical ground reaction forces during impact were analyzed.Results: We found significant main effects for developmental stage. Children demonstrated greater knee valgus and less hip flexion at initial contact and at maximum vertical force, less knee flexion at maximum vertical force, greater maximum vertical force and impulse, and a shorter time to maximum vertical force than the adults. No sex differences were found among the biomechanical variables measured.Conclusions: The presence of significant biomechanical differences between children and adults suggests that physical development influences landing patterns. Sex does not appear to influence landing patterns during a 50% maximum vertical jump landing. These findings add to the body of knowledge regarding developmental and sex comparisons in a functional landing task. PMID- 15902319 TI - On-the-Field Resistance-Tubing Exercises for Throwers: An Electromyographic Analysis. AB - Context: Athletes who throw commonly use rubber-tubing resistance exercises in the field setting to assist with warm-up before throwing. Yet no researchers have described which muscles are being activated or which exercises are most effective during rubber-tubing exercises used by throwers for warm-up.Objective: To describe the effectiveness of 12 rubber-tubing resistance exercises commonly used by throwers in activating the shoulder muscles important for throwing.Design: Descriptive research design.Setting: An applied biomechanics research laboratory.Patients or Other Participants: Fifteen physically active male subjects with no history of shoulder injury.Main Outcome Measure(s): Subjects randomly performed 12 rubber-tubing resistance exercises while we assessed muscle activation of the subscapularis, supraspinatus, teres minor, and rhomboid major by indwelling electromyography. Activation of the sternal portion of the pectoralis major, anterior deltoid, middle deltoid, latissimus dorsi, serratus anterior, biceps brachii, triceps brachii, lower trapezius, and infraspinatus muscles was assessed by surface electromyography.Results: Performance of 7 exercises (external rotation at 90 degrees of abduction, throwing deceleration, humeral flexion, humeral extension, low scapular rows, throwing acceleration, and scapular punch) resulted in the highest level of muscle activation of all muscles tested.Conclusions: These 7 exercises exhibited moderate activation (>20% maximal voluntary isometric contraction) in each muscle of the rotator cuff, the primary humeral movers, and the scapular stabilizer muscles. The results suggest that these exercises are most effective in activating the muscles important to the throwing motion and may be beneficial for throwers during their prethrowing warm up routine. PMID- 15902320 TI - Clinical Measures of Shoulder Mobility in the Professional Baseball Player. AB - Context: Professional baseball players must achieve a delicate balance between shoulder mobility and stability to attain optimal sports performance. The sport specific demands of repetitive overhead throwing may result in an altered mobility-stability relationship.Objective: To evaluate clinical measures of shoulder mobility in professional baseball players in order to examine differences between the throwing and the nonthrowing shoulders and to describe chronic adaptations to throwing.Design: Descriptive.Setting: The athletic training room at Maryvale Baseball Park, Phoenix, AZ.Patients or Other Participants: Twenty-seven professional baseball players (20 pitchers, 7 position players; age = 20 +/- 1.6 years, height = 190.5 +/- 4.8 cm, mass = 91.6 +/- 9.6 kg) with no previous history of shoulder or elbow injury.Main Outcome Measure(s): We recorded scapular upward rotation at 4 levels of humeral elevation in the scapular plane (rest, 60 degrees , 90 degrees , 120 degrees ); posterior shoulder tightness; and passive, isolated glenohumeral joint internal and external range of motion.Results: Scapular upward rotation was significantly greater in the throwing shoulder (14.2 +/- 6.5 degrees ) than in the nonthrowing shoulder (10.6 +/- 6.1 degrees ) at 90 degrees of humeral elevation (P = .04). We observed no statistically significant difference in posterior shoulder tightness between the throwing (30.2 +/- 4.6 cm) and the nonthrowing (28.0 +/- 4.8 cm) shoulder (P = .09). In addition, the throwing shoulder exhibited a statistically significant decrease in isolated glenohumeral internal rotation (56.6 +/- 12.5 degrees ) compared with the nonthrowing shoulder (68.6 +/- 12.6 degrees ) (P = .001), with a concomitant increase in isolated glenohumeral external rotation (throwing = 108.9 +/- 9.0 degrees , nonthrowing = 101.9 +/- 5.9 degrees , P = .0014). An analysis of the total arc of motion (internal rotation + external rotation) revealed no statistically significant difference between sides (P = .15).Conclusions: The throwing shoulder exhibited significant differences in scapular and glenohumeral mobility compared with the nonthrowing shoulder. Further research is necessary to determine the relation of these adaptive changes, if any, to shoulder injury and disability. PMID- 15902321 TI - Functional-Performance Deficits in Volunteers With Functional Ankle Instability. AB - Context: Although functional-performance tests are dynamic measures used to assess general lower body function, studies investigating these tests for ankle instability have yielded conflicting results.Objective: To determine if a relationship exists between a measure of functional ankle instability and deficits in functional performance.Design: A case-control study correlating subject performance on a set of lower extremity functional-performance tests with a measure of ankle instability.Setting: University athletic training research laboratory.Patients or Other Participants: We recruited 60 participants (43 females, 17 males, age = 22.4 +/- 4.9 years, height = 169.9 +/- 9.7 cm, mass = 72.6 +/- 16.3 kg; 42 injured, 18 uninjured) to participate in the study. Six questions were used to determine if functional ankle instability was present in each participant. A point was added for each yes response to produce an index that represents a continuous variable of functional ankle instability.Main Outcome Measure(s): Four unilateral hopping tests were used in this study: figure of-8 hop, side hop, up-down hop, and single hop. For the first 3 tests, the total time was recorded with a handheld stopwatch to the nearest 0.01 second; for the single hop-for-distance test, the distance was recorded to the nearest 0.01 m. Correlations were computed with the functional ankle instability index and each of the 4 functional-performance tests.Results: No relationship was revealed between the functional ankle instability index and single hop for distance or up down hop, with r values of -.008 and .245, respectively. A significant relationship was found between the functional ankle instability index and the side hop (r = .35, P /=10 mm), neutral (5-9 mm), or supinated (or= 50% diameter stenosis) was found in 56/84 patients, 30 patients had multivessel disease (14 in the Dip-Atro and 16 in the Dip-Dob group; p>0.05). Sensitivity, specificity, diagnostic accuracy, positive predictive value, and negative predictive value were 92.9%, 78.6%, 88.1%, 89.7%, and 84.6% versus 89.3%, 92.9%, 90.5%, 96.2%, and 81.2% (p>0.05, Dip-Dob vs. Dip-Atro). CONCLUSION: There was no significant difference in feasibility, safety and diagnostic accuracy between the Dip-Dob and Dip-Atro test. The Dip-Dob test showed more pronounced ability to provoke myocardial ischemia than the Dip-Atro test, expressed as a higher delta WMSI. Both tests confirmed high diagnostic accuracy of pharmacological stress echocardiography in hypertensive patients. PMID- 15902373 TI - [Percutaneous transplantation of autologous myoblasts in ischemic cardiomyopathy]. AB - BACKGROUND AND PURPOSE: Cell transplantation is emerging as a novel approach for the treatment of end-stage cardiac disease. In contrast to most human studies using intramyocardial injection of myoblasts during coronary artery bypass grafting (CABG) or left ventricular assist device implantation, the authors investigated both safety and feasibility of transcatheter transplantation of autologous skeletal myoblast as a standalone procedure in six patients with ischemic heart failure, and compared them to six control patients matched for demographic and clinical characteristics. METHODS AND RESULTS: Skeletal myoblast transplantation by catheter-based injection was technically successful in all six patients with no complications; 19+/-10 injections were performed/patient corresponding to 210 x 10(6)+/-150 x 10(6) cells/patient. Postinterventional Holter monitoring and ICD memory check documented three episodes of ventricular tachycardia in two patients after myoblast implantation, one at 30 days in patient 1, and two at 27 and 41 days in patient 2. Patient 1, although asymptomatic, was subsequently subjected to oral amiodarone since he refused an ICD; in patient 2 each tachycardia was terminated by a previously implanted ICD. Both patients were followed for 6 months without any evidence of repeat ventricular arrhythmia. Matched control patients revealed one episode of ventricular tachycardia in three patients each of which was aborted by ICD discharge within 6 months of observation. None of the documented ventricular tachycardias in both groups occurred in relation to any new myocardial necrosis which was excluded by ECG and cardiac enzymes. Left ventricular ejection fraction (LVEF) rose from 24.3+/-6.7% at baseline to 33.2+/-10.2% 6 months after myoblast implantation (p=0.02 vs. baseline); in matched controls LVEF decreased from 24.7+/-4.6% to 22.2+/-6.2% (p<0.05 vs. myoblasttreated group at 6 months). Moreover, the 6-min walk test revealed an improvement from 371+/-49 m to 493+/-86 m 6 months after implantation (p=0.003 vs. baseline and p=0.015 vs. controls), whereas matched controls were unchanged at 360+/-24 m and 369+/-26 m, respectively. Accordingly, NYHA functional class improved from 3.17+/-0.41 to 1.67+/-0.82 within 6 months of myoblast implantation (p=0.001 vs. baseline and p=0.01 vs. controls), while NYHA class remained unchanged at 3+/-0 in matched controls. CONCLUSION: Transcatheter transplantation of autologous skeletal myoblasts for severe left ventricular dysfunction in postinfarction patients is feasible, safe and promising and, thus, warrants the scrutiny of larger randomized double-blind multi-center trials with longer follow-up surveillance. PMID- 15902374 TI - [Mis-diagnosed Morbus Fallot]. PMID- 15902375 TI - [Echocardiography in the diagnosis of endocarditis: transthoracic or transesophageal?]. AB - Echocardiography is an important tool for diagnosis and risk assessment in patients with infective endocarditis. In addition to the identification of typical vegetations, echocardiography is crucial for the evaluation of valvar regurgitations, and the early diagnosis of complications. Transesophageal echocardiography is, in nearly every aspect, except with concern to right-sided endocarditis, superior to the transthoracic approach. Thus, transesophageal echocardiography represents, in patients with substantial clinical suspicion for an active infection of endocardial structures, the currently most sensitive diagnostic tool. PMID- 15902376 TI - [Acute ST-segment-elevation myocardial infarction. Current treatment situation for patients in Germany]. PMID- 15902377 TI - [Has the incidence of end-stage renal disease increased in diabetic patients? A center-based longitudinal study over 10 years]. AB - BACKGROUND AND PURPOSE: Worldwide, the incidence of diabetic patients requiring renal replacement therapy seems to be on the rise. Whether this assumption holds true also for Germany was investigated on the basis of three long-standing centers for dialysis in the catchment area of Dusseldorf. PATIENTS AND METHODS: All patients entering long-term dialysis between 1990 and 1999 were analyzed in retrospect according to their charts. The following items were evaluated as documented: diagnosis, type, and duration of diabetes mellitus; diabetic neuropathy and retinopathy; amputations; causes of hemodialysis. RESULTS: Between 1990 and 1999, a total of 657 patients (379 men, 278 women) entered hemodialysis. The mean entering rate remained at 67/year between 1990 and 1994 and at 64/year between 1995 and 1999 (n.s.). The mean age of all patients at entering dialysis increased significantly from 59 years between 1990 and 1994 to 62 years between 1995 and 1999 (p = 0.0069). A total of 243 patients were known diabetics; their mean entering rate of 24/year between 1990 and 1994 continued at 25/year between 1995 and 1999 (n.s.). The mean age of the diabetic patients increased from 62 years between 1990 and 1994 to 66 years between 1995 and 1999 (p = 0.0238). However, the proportion of diabetic patients entering dialysis remained unchanged at approximately 37% over the 10-year time period (Cochran-Armitage trend test; p = 0.1568). Of all diabetic patients, 92 had proven diabetic retinopathy, 33 were amputated, and 38 had neuropathy. In 37% the medical records did not mention any etiology of end-stage renal disease in diabetic patients, 29% were nondiabetic causes, and 34% were said to be diabetic nephropathy. CONCLUSION: The incidence of patients with end-stage renal disease entering hemodialysis has not changed between 1990 and 1999, and the proportion of diabetic patients has remained the same. However, the patients' age at entering hemodialysis has increased significantly between 1990 and 1999. As the documentation in the patients' records was unsatisfactory with regard to diabetes history and causes for dialysis, the results of the present study need to be interpreted with caution. PMID- 15902378 TI - [Bronchodilators in chronic obstructive pulmonary disease (COPD)]. AB - Bronchodilators form the foundation of the pharmacotherapy of patients with chronic obstructive pulmonary disease (COPD). Scores of information from numerous large-scale clinical trials, mechanistic differences between classes of bronchodilators, and anti-inflammatory/bronchodilator fixed combinations make the decision what compound primarily to prefer in COPD treatment a challenge. In this review of large, double-blind, clinical trials with anticholinergic drugs, long- and short-acting beta(2)-agonists, xanthines and different application forms and combination of these compounds will be examined for clinical efficacy. The following practical objectives were accepted to define effective disease management: improvement of lung function, physical parameters such as 6-min walking distance, reduction of exacerbation rate and severity, improvement of quality of life and dyspnea score. Based on this review, inhalation therapy with a long-acting bronchodilator such as tiotropium, formoterol or salmeterol is proposed for early treatment algorithm. The combination of an anticholinergic compound and a long-acting bronchodilator may have an additive effect on bronchodilatation. The addition of inhaled corticosteroids is only recommended in stages III and IV. Besides, pharmacotherapy of COPD should always be flanked by smoking prevention programs, and supportive therapy, if indicated in severe disease stages. PMID- 15902379 TI - [Costs of ambulatory care for RA patients in Germany]. AB - BACKGROUND: In 2000, the Regional Cooperative Center for Rheumatic Conditions in Hannover, Germany, initiated a project to introduce and evaluate a case-based clinical quality management system to enhance the quality of care for patients with rheumatoid arthritis. Building on that, it was possible to combine all resource uses, as paid by the sickness funds, including those in ambulatory care. METHODS: The purpose of this study was to reach average "per-visit prices" for single physician visits, not only for rheumatologists, but also for other specialists as well as generalists. Those visits all had to be connected to the rheumatic condition of the patients. All ambulatory care visits for 1 year (2001) were evaluated. RESULTS: Due to the high number of physician visits of the 338 patients with clinically assured rheumatoid arthritis included in the study (rheumatologist visits 4,488, generalist visits 3,901), statistically significant subgroups could be built. The average costs per visit associated with rheumatoid arthritis are: rheumatologist euro 22.71; generalist euro 8.02; as well as other specialties ranging from euro 5.81 to euro 19.48. The low price for generalists care is due to the selection of the cohort, as all patients are under constant specialist care. The prices were further broken down to certain subgroups of care. CONCLUSION: The prices are calculated under the premises that no budgetary constraints apply to ambulatory care. This is not the reality in Germany. Hence, looking at the average frequency of rheumatologist visits in this cohort (13.2/year), the conclusion has to be that not all rheumatologic ambulatory care is being covered by the reimbursement system within the Statutory Health Insurance in Germany. PMID- 15902380 TI - [The patient with syncope]. PMID- 15902381 TI - [Severe erythrodermic psoriasis in a patient with 22q11 deletion syndrome]. AB - BACKGROUND: Erythrodermic psoriasis is a severe manifestation of psoriasis and can be triggered by several factors. CASE REPORT: A 35-year-old man was admitted with severe, almost generalized exfoliative, oozing erythrodermic psoriasis, fever, and cramping of hands and legs. He was under systemic treatment with acitretin. Laboratory examination revealed a marked hypocalcemia as a consequence of primary hypopara thyroidism as well as hypalbuminemia. After normalization of the serum calcium and albumin levels, cutaneous symptoms and fever rapidly improved. No infectious etiology could be found. Hypoparathyroidism together with a right-sided aorta was caused by a 22q11 deletion syndrome. CONCLUSION: Epidermal cell proliferation and formation of intercellular junctional components of the epidermis are strongly calcium-dependent. Furthermore, carriers like albumin are necessary for the transportation of acitretin in the peripheral tissue. This case report suggests that calcium can be involved in psoriasis pathogenesis at least in a subgroup of patients and that systemic retinoids exhibit insufficient effectiveness under low serum albumin levels. PMID- 15902382 TI - [Acute coronary syndrome in a 17-year-old female with systemic lupus erythematosus]. AB - CASE REPORT: A 17-year-old female was admitted to the hospital with epigastric pain radiating into the chest and neck. The patient had a known, long-standing history of systemic lupus erythematosus (SLE) with predominantly renal involvement. Diagnostic examinations including left heart catheterization showed acute myocardial infarction based on a coronary three-vessel disease. Percutaneous transluminal coronary angioplasty (PTCA) with dilatation of the left marginal branch and the left posterolateral branch was performed. 8 weeks after PTCA, the patient was admitted again showing occlusion of the same coronary artery. A coronary stent was implanted which, after 8 weeks, also showed signs of occlusion. This case report describes premature myocardial infarction in a young woman with SLE and addresses and discusses the problem that myocardial infarction in SLE can be caused by atherosclerosis and/or arteritis of the coronary arteries. PMID- 15902383 TI - [Nosocomial infections]. PMID- 15902384 TI - [Hypereosinophilia as the leading symptom in a patient with mastocytosis]. PMID- 15902385 TI - [Palliative sedation--an alternative to euthanasia in intractable pain at the end of life?]. PMID- 15902386 TI - [Sentinel lymphadenectomy as a staging procedure]. PMID- 15902387 TI - [60 year old patient with soft tissue infection of the right leg]. AB - Group A streptococcal necrotizing fasciitis is a rare disease associated with high mortality. Since severe toxic shock syndrome is a common complication, only immediate and aggressive surgical intervention, adequate antimicrobial therapy and supportive intensive care can be life-saving. We report about successful treatment of a 60-year old patient with necrotizing fasciitis and multiple organ failure. PMID- 15902388 TI - The genetics of atopic dermatitis: recent findings and future options. AB - Atopic dermatitis (AD) is a chronic pruritic skin disease affecting up to 15% of children in industrialized countries. AD belongs to the group of allergic disorders that include food allergy, allergic rhinitis, and asthma. A multifactorial background for AD has been suggested, with genetic as well as environmental factors influencing disease development. Genome-wide screens for AD have been completed in four different populations to date. Interestingly, the susceptibility regions identified for AD show little overlap with asthma susceptibility regions, suggesting that, at least in part, separate genes might be involved in the pathogenesis of the different atopic disorders. Instead, some of the identified regions overlap with susceptibility regions for psoriasis, another chronic skin disease. Thus, genes expressed in the skin might play an important role in AD pathogenesis, in addition to genes influencing atopic diatheses. Although no veritable "AD gene" has been identified by positional cloning to date, examples from other complex genetic disorders such as asthma show that this goal is likely to be reached in the near future. Candidate gene studies, on the other hand, have identified 19 genes that were shown to be associated with AD in at least one study. The results of genome-wide screens as well as candidate gene studies are evaluated here in detail. PMID- 15902389 TI - Topical application of plasmid DNA to mouse and human skin. AB - Gene expression following direct injection of naked plasmid DNA into the skin has been demonstrated in the past. Topical application of plasmid DNA represents an attractive route of gene delivery. If successful, it would have great prospects in skin gene therapy since it is painless and easy to apply. In this study, we analyzed the expression of plasmid DNA in vivo and in vitro following topical application of plasmid DNA in various liposomal spray formulations. Therefore, different concentrations of plasmid DNA expressing enhanced green fluorescent protein (pEGFP-N1) were sprayed onto mouse or human skin once daily for three consecutive days and compared with direct injection. Gene expression was assessed 24 h after the final topical application of various liposomal DNA formulations. The results showed that EGFP mRNA and protein were detectable by RT-PCR and Western blot, respectively. However, epicutaneously applied EGFP plasmid DNA did not lead to microscopically detectable EGFP protein, when assessed by confocal laser microscopy or fluorescence-activated cell sorting in contrast to about 4% of fluorescent keratinocytes following intradermal injection. In an in vivo mouse model, the application of pEGFP-N1 DNA led to the generation of GFP-specific antibodies. These results indicate that topical spray application of pEGFP-N1 liposomal DNA formulations is a suitable method for plasmid DNA delivery to the skin, yielding limited gene expression. This spray method may thus be useful for DNA vaccination. To increase its attractiveness for skin gene therapy, the improvement of topical formulations with enhanced DNA absorption is desirable. PMID- 15902390 TI - [High-dose intravenous immunoglobulins in the treatment of multiple sclerosis. An update]. AB - The immunomodulatory treatment of multiple sclerosis (MS) with high-dose intravenous immunoglobulins (IVIg) has been discussed with some controversy in the context of evidence-based medicine. The recent publication of eight trials investigating several aspects of MS has shed some more light on the role of IVIg treatment in MS. Here we summarize and critically discuss the new data in the context of previous studies on this treatment. In relapsing-remitting MS, IVIg remain a second-line treatment when other licensed treatments are not possible. Currently there is no role for IVIg in secondary progressive MS. Similarly, the use of IVIg during an acute relapse shows no benefit in addition to standard steroid treatment. The initiation of IVIg therapy after a clinically isolated syndrome has delayed the occurrence of definite MS, and this may become a new indication. Furthermore, previous data suggesting that IVIg can reduce the incidence of postpartal relapses have been substantiated. However, those trials unfortunately lack appropriate internal control groups. By and large, previous recommendations for the use of IVIg in MS are supported by the new data. PMID- 15902391 TI - [The causes of statin induced myopathies]. PMID- 15902392 TI - [Bilateral pallidal infarction in sleep apnea syndrome]. AB - Patients suffering from sleep apnea syndrome are known to be at higher risk of cardiac and cerebral infarction. In this case report, we describe bilateral pallidal lesions, which are normally seen after sudden asphyxia due to cardiac arrest or carbon monoxide intoxication. Some epidemiological and pathophysiological observations are cited. PMID- 15902393 TI - [Multiple ischemic vertebrobasilar lesions in temporal arteritis]. AB - We report a 73-year-old patient with giant cell arteritis who presented with right arm paresis and dysarthria. Vasculitis of the vertebral arteries induced multiple ischemic cerebral lesions in the vertebrobasilar territory, ultimately leading to death despite intensive immunosuppressive therapy. This case illustrates that prompt relief from symptoms of giant cell arteritis upon steroid treatment does not correspond to a remission of the vasculitic process itself. Clinicians should therefore be aware of the possible occult involvement of the vertebral or other major arteries. PMID- 15902394 TI - [Endovascular occlusion of the basilar artery for the treatment of dissecting and dysplastic fusiform aneurysms]. AB - Dissecting aneurysms of the basilar artery trunk frequently affect young adults. Fusiform shape and narrowing of the proximal parent artery are typical features. Changes in aneurysm size and geometry may be observed more rapidly than in atherosclerotic or dysplastic aneurysms. Dissecting aneurysms carry a significant risk of rupture. Thrombotic or embolic occlusion of small pontine branches may cause ischemic symptoms. Sufficiently large aneurysms compress the adjacent brainstem. The operative treatment of these aneurysms is associated with unacceptable risks. At least one posterior communicating artery with normal calibre together with the ipsilateral P1 segment needs to provide adequate collateral flow to the upper basilar artery to allow endovascular coil occlusion of the segment that is affected by the dissection and/or fusiform aneurysmal dilatation. Four illustrative cases of endovascular coil occlusion of the basilar artery for the treatment of fusiform aneurysms are presented and discussed. PMID- 15902395 TI - AB-QTL analysis in spring barley. I. Detection of resistance genes against powdery mildew, leaf rust and scald introgressed from wild barley. AB - The objective of this study was to map new resistance genes against powdery mildew (Blumeria graminis f. sp. hordei L.), leaf rust (Puccinia hordei L.) and scald [Rhynchosporium secalis (Oud.) J. Davis] in the advanced backcross doubled haploid (BC2DH) population S42 derived from a cross between the spring barley cultivar 'Scarlett' and the wild barley accession ISR42-8 (Hordeum vulgare ssp. spontaneum). Using field data of disease severity recorded in eight environments under natural infestation and genotype data of 98 SSR loci, we detected nine QTL for powdery mildew, six QTL for leaf rust resistance and three QTL for scald resistance. The presence of the exotic QTL alleles reduced disease symptoms by a maximum of 51.5, 37.6 and 16.5% for powdery mildew, leaf rust and scald, respectively. Some of the detected QTL may correspond to previously identified qualitative (i.e. Mla) and to quantitative resistance genes. Others may be newly identified resistance genes. For the majority of resistance QTL (61.0%) the wild barley contributed the favourable allele demonstrating the usefulness of wild barley in the quest for resistant cultivars. PMID- 15902396 TI - Genetic and physical mapping of the grapevine powdery mildew resistance gene, Run1, using a bacterial artificial chromosome library. AB - Resistance to grapevine powdery mildew is controlled by Run1, a single dominant gene present in the wild grapevine species, Muscadinia rotundifolia, but absent from the cultivated species, Vitis vinifera. Run1 has been introgressed into V. vinifera using a pseudo-backcross strategy, and genetic markers have previously been identified that are linked to the resistance locus. Here we describe the construction of comprehensive genetic and physical maps spanning the resistance locus that will enable future positional cloning of the resistance gene. Physical mapping was performed using a bacterial artificial chromosome (BAC) library constructed using genomic DNA extracted from a resistant V. vinifera individual carrying Run1 within an introgression. BAC contig assembly has enabled 20 new genetic markers to be identified that are closely linked to Run1, and the position of the resistance locus has been refined, locating the gene between the simple sequence repeat (SSR) marker, VMC4f3.1, and the BAC end sequence-derived marker, CB292.294. This region contains two multigene families of resistance gene analogues (RGA). A comparison of physical and genetic mapping data indicates that recombination is severely repressed in the vicinity of Run1, possibly due to divergent sequence contained within the introgressed fragment from M. rotundifolia that carries the Run1 gene. PMID- 15902397 TI - Quantitative trait loci for cell-wall components in recombinant inbred lines of maize (Zea mays L.) I: stalk tissue. AB - Maize silage is a significant energy source for animal production operations, and the efficiency of the conversion of forage into animal mass is an important consideration when selecting cultivars for use as feed. Fiber and lignin are negatively correlated with digestibility of feed, so the development of forage with reduced levels of these cell-wall components (CWCs) is desirable. While variability for fiber and lignin is present in maize germplasm, traditional selection has focused on the yield of the ear rather than the forage quality of the whole plant, and little information is available concerning the genetics of fiber and lignin. The objectives of this study were to map quantitative trait loci (QTLs) for fiber and lignin in the maize stalk and compare them with QTLs from other populations. Stalk samples were harvested from 191 recombinant inbred lines (RILs) of B73 (an inbred line with low-to-intermediate levels of CWCs) x De811 (an inbred line with high levels of CWCs) at two locations in 1998 and one in 1999 and assayed for neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL). The QTLs were detected on nine chromosomes, mostly clustered in concordance with the high genetic correlations between NDF and ADF. Adjustment of NDF for ADF and ADF for ADL revealed that most of the variability for CWCs in this population is in ADF. Many of the QTLs detected in this study have also been detected in other populations, and several are linked to candidate genes for cellulose or starch biosynthesis. The genetic information obtained in this study should be useful to breeding efforts aimed at improving the quality of maize silage. PMID- 15902398 TI - Genome-wide analysis of the frequency and distribution of crossovers at male and female meiosis in Sinapis alba L. (white mustard). AB - We present the first genetic linkage maps of Sinapis alba (white mustard) and a rigorous analysis of sex effects on the frequency and distribution of crossovers at meiosis in this species. Sex-averaged maps representing recombination in two highly heterozygous parents were aligned to give a consensus map consisting of 382 loci defined by restriction fragment length polymorphisms and arranged in 12 linkage groups with no unlinked markers. The loci were distributed in a near random manner across the genome, and there was little evidence of segregation distortion. From these dense maps, a subset of spaced informative markers was used to establish recombination frequencies assayed separately in male and female gametes and derived from two distinct genetic backgrounds. Analyses of 746 gametes indicated that recombination frequencies were greater in male gametes, with the greatest differences near the ends of linkage groups. Genetic background had a lesser effect on recombination frequencies, with no discernible pattern in the distribution of such differences. The possible causes of sex differences in recombination frequency and the implications for plant breeding are discussed. PMID- 15902399 TI - Mapping and tagging of seed coat colour and the identification of microsatellite markers for marker-assisted manipulation of the trait in Brassica juncea. AB - Microsatellite marker technology in combination with three doubled haploid mapping populations of Brassica juncea were used to map and tag two independent loci controlling seed coat colour in B. juncea. One of the populations, derived from a cross between a brown-seeded Indian cultivar, Varuna, and a Canadian yellow-seeded line, Heera, segregated for two genes coding for seed coat colour; the other two populations segregated for one gene each. Microsatellite markers were obtained from related Brassica species. Three microsatellite markers (Ra2 A11, Na10-A08 and Ni4-F11) showing strong association with seed coat colour were identified through bulk segregant analysis. Subsequent mapping placed Ra2-A11 and Na10-A08 on linkage group (LG) 1 at an interval of 0.6 cM from each other and marker Ni4-F11 on LG 2 of the linkage map of B. juncea published previously (Pradhan et al., Theor Appl Genet 106:607-614, 2003). The two seed coat colour genes were placed with markers Ra2-A11 and Na10-A08 on LG 1 and Ni4-F11 on LG 2 based on marker genotyping data derived from the two mapping populations segregating for one gene each. One of the genes (BjSC1) co-segregated with marker Na10-A08 in LG 1 and the other gene (BjSC2) with Ni4-F11 in LG 2, without any recombination in the respective mapping populations of 130 and 103 segregating plants. The identified microsatellite markers were studied for their length polymorphism in a number of yellow-seeded eastern European and brown-seeded Indian germplasm of B. juncea and were found to be useful for the diversification of yellow seed coat colour from a variety of sources into Indian germplasm. PMID- 15902400 TI - Glucagon-like peptide-1 protects beta cells from cytokine-induced apoptosis and necrosis: role of protein kinase B. AB - AIMS/HYPOTHESIS: The gut hormone glucagon-like peptide-1 (GLP-1) decreases beta cell apoptosis in a protein kinase B (PKB)-dependent fashion, and increases islet cell mass and function in vivo. In contrast, cytokines induce beta cell apoptosis, leading to decreased islet mass and type 1 diabetes. In the present study we used rat INS-1E beta cells and primary rat islet cells to examine the potential role of PKB as a mediator of the effect of GLP-1 on cytokine-induced apoptosis. METHODS: Cell viability was determined by MTT assay, and apoptosis and necrosis by Hoechst 33342-propidium iodide staining. Immunoblot analysis was used to detect changes in protein expression, including active (phosphorylated) and total PKB, phosphorylated and total glycogen synthase kinase-3beta, activated caspase-3 and inducible nitric oxide synthase. Reactive oxygen species were determined by 1,7-dichlorofluorescein (DCF) analysis, and mutant forms of PKB were introduced into cells using adenoviral vectors. RESULTS: Incubation of INS 1E cells with cytokines (IL-1beta, TNF-alpha and interferon-gamma; 10-50 ng/ml) for 18 h significantly decreased cell viability (by 44%, p<0.001), cell proliferation (by 80%, p<0.001), and activation of PKB (by 67%, p<0.001). Pre treatment with exendin-4 (10(-7) mol/l), a long-acting GLP-1 receptor agonist, partially protected the cells against cytokine-induced toxicity (p<0.01) in association with a reduction in cytokine-induced inhibition of PKB phosphorylation (p<0.05). Exendin-4 pre-treatment did not change cell proliferation. Cytokine treatment increased apoptosis (by 156%, p<0.05) and necrosis (from undetectable to 2.6% of cells). These increases were both reduced by pre-treatment with exendin-4 (p<0.05-0.01). Furthermore, cytokine-induced apoptosis and necrosis were significantly increased in cells infected with kinase dead PKB (p<0.05), and the protective effect of exendin-4 on both parameters was fully abolished in these cells. Similar changes were observed in primary islet cells. In parallel with these changes, exendin-4 decreased the cytokine-induced activation of caspase-3 (by 46%, p<0.05), and decreased levels of inducible nitric oxide synthase (by 71%, p<0.05) and reactive oxygen species (by 27%, p<0.05). CONCLUSIONS/INTERPRETATION: The results of our study indicate that GLP-1 plays a protective role against cytokine-induced apoptosis and necrosis in beta cells through a PKB-dependent signalling pathway. PMID- 15902401 TI - Relationship between the incidence of type 1 diabetes and maternal enterovirus antibodies: time trends and geographical variation. AB - AIMS/HYPOTHESIS: We have previously observed an inverse correlation between the incidence of type 1 diabetes and enterovirus infections in the background population. The aim of this study was to analyse whether maternal enterovirus antibody status, which reflects both the frequency of enterovirus infections and the protection conferred by the mother on the offspring, also correlates with the incidence of type 1 diabetes. METHODS: Maternal enterovirus antibodies were analysed from serum samples taken from pregnant women between 1983 and 2001 in Finland and Sweden using enzyme immunoassay and neutralisation assays. Comparable samples were also taken between 1999 and 2001 in countries with a lower incidence of diabetes (Estonia, Germany, Hungary, Israel, Lithuania, Russia). RESULTS: A clear decrease was observed in maternal enterovirus antibody levels over the past 20 years (p<0.0001). The frequency of enterovirus antibodies was higher in countries with a low or intermediate incidence of type 1 diabetes compared with high-incidence countries (p<0.0001). CONCLUSIONS/INTERPRETATION: These findings are in line with our previous observations supporting the hypothesis that a low frequency of enterovirus infection in the background population increases the susceptibility of young children to the diabetogenic effect of enteroviruses. PMID- 15902402 TI - Serum high molecular weight complex of adiponectin correlates better with glucose tolerance than total serum adiponectin in Indo-Asian males. AB - AIMS/HYPOTHESIS: It is well established that total systemic adiponectin is reduced in type 2 diabetic subjects. To date most studies have been concerned with the singular full-length protein or proteolytically cleaved globular domain. It is, however, apparent that the native protein circulates in serum as a lower molecular weight hexamer and as larger multimeric structures of high molecular weight (HMW). In this study we address the clinical significance of each form of the protein with respect to glucose tolerance. METHODS: Serum was obtained from 34 Indo-Asian male subjects (BMI 26.5+/-3.1; age 52.15+/-10.14 years) who had undertaken a 2-h oral glucose tolerance test. An aliquot of serum was fractionated using velocity sedimentation followed by reducing SDS-PAGE. Western blots were probed for adiponectin, and HMW adiponectin as a percentage of total adiponectin (percentage of higher molecular weight adiponectin [S(A)] index) was calculated from densitometry readings. Total adiponectin was measured using ELISA; leptin, insulin and IL-6 were determined using ELISA. RESULTS: Analysis of the cohort demonstrated that total adiponectin (r = 0.625, p = 0.0001), fasting insulin (r = -0.354, p = 0.040) and age (r = 0.567, p = 0.0001) correlated with S(A). S(A) showed a tighter, inverse correlation with 2-h glucose levels (r = 0.58, p = 0.0003) than total adiponectin (r = -0.38, p = 0.0001). CONCLUSIONS/INTERPRETATION: This study demonstrates the importance of the S(A) index as a better determinant of glucose intolerance than measurements of total adiponectin. Our findings suggest that HMW adiponectin is the active form of the protein. PMID- 15902403 TI - A community intervention for early identification of first episode psychosis: impact on duration of untreated psychosis (DUP) and patient characteristics. AB - OBJECTIVE: The aim of this study was to assess the impact of a community case identification program on duration of untreated psychosis (DUP) (a measure of delay in treatment) and characteristics of patients entering treatment for a first episode of psychosis. METHOD: Using a quasi-experimental historical control design, patients within a defined geographic catchment area who met DSM-IV criteria for a first episode of a psychotic disorder (FEP) were assessed on a number of demographic and clinical variables including DUP, length of prodromal period and symptoms at initial presentation, for 2 years prior to and 2 years after the introduction of a community-wide Early Case Identification Program (ECIP). The ECIP was designed to promote early recognition and referral of individuals with a FEP from any possible source of referral including self referrals. Treatment interventions offered were the same throughout the two phases. RESULTS: In all, 88 and 100 patients met criteria respectively in phases I and II. There were no significant differences in rates of treated incidence or DUP between the two phases. Patients recruited in phase II had significantly longer prodromal periods and higher level of psychotic and disorganization symptoms. There were no differences in level of negative symptoms or pre-morbid adjustment. CONCLUSION: A community-wide approach to early case identification may not be the most effective way to reduce delay in treatment of psychosis, but may bring into treatment patients who have been ill for long periods of time and have a higher level of psychopathology. A more targeted approach directed at primary care and emergency services may achieve different results in reducing delay in treatment. PMID- 15902404 TI - Duration of untreated psychotic illness: the role of premorbid social support networks. AB - BACKGROUND: A lengthy delay often exists between the onset of psychotic symptoms and the start of appropriate treatment. However, the causes of this long delay remain poorly understood, and there is a need to search for the factors involved in such a delay in order to reduce the time of untreated psychosis. This study aimed at examining the influence of premorbid social networks on the duration of untreated psychotic illness. METHOD: One hundred subjects with a first episode of schizophrenia or related psychotic disorders never treated with antipsychotics made up the study sample. Social support was assessed by means of the Sturtees's social support scale that comprises two subscales measuring close and diffuse social support. Duration of untreated illness was assessed according to three definitions: duration of untreated unspecific symptoms, duration of untreated psychotic symptoms, and duration of untreated continuous psychotic symptoms. We also examined the effect of putative confounding factors such as gender, residence (urban-rural), age at illness onset, years of education, and parental socio-economic status. RESULTS: Correlational analysis showed that poor diffuse social support, but not poor close social support, predicted long duration of untreated illness according to the three definitions; this association being mainly due to poor work/academic support. Logistic regression analysis confirmed such an association, but it was limited to duration of continuous psychotic symptoms (unadjusted OR=3.44, 95% CI=1.51-7.83); this association persisted after adjusting for the confounding variables (adjusted OR=3.39, 95% CI=1.39-8.29). We also found that subjects with low socio-economic status were depending on the definition of duration of untreated illness considered, between 2.7 and 4.3 times more likely to present with a long duration of untreated illness. CONCLUSION: Both poor diffuse social support and a low socio-economic status seem to be relevant factors of a prolonged duration of untreated psychosis. PMID- 15902405 TI - The impact of family experience on the duration of untreated psychosis (DUP) in Hong Kong. AB - BACKGROUND: Previous family experience of psychotic illness may play an important role in whether and when a patient seeks help in first-episode psychosis. This study investigated the relationship between family experience of psychosis and the duration of untreated psychosis in a prospective sample of first-episode psychosis patients in Hong Kong. We also studied the effects of pre-morbid adjustment, educational level, living alone, and mode of onset as potential determinants of the duration of untreated psychosis (DUP). METHODS: A total of 131 first-episode psychosis patients in Hong Kong were recruited in a study of the DUP and related factors. The Interview for the Retrospective Assessment of the Onset of Schizophrenia (IRAOS) was used to measure the DUP and to provide a structured assessment of family history, educational level, household arrangement, and mode of onset. RESULTS: Previous family experience of psychiatric illness (the presence of another family member who has been receiving psychiatric treatment) and an acute mode of onset were significant predictors of a shorter DUP. Educational level had a modest effect on its own, but was not significant in the binary logistic regression model. Living alone had a moderate effect size, but was non-significant, possibly because of the small proportion of single-person households in the sample. The symptom profile, pre-morbid adjustment, and other demographic factors were not significantly related to the DUP. CONCLUSION: In addition to the mode of onset, previous family experience plays an important role in the presentation of early psychosis. Educational efforts that target the family should be an important part of any strategy for the early detection of psychosis. PMID- 15902406 TI - Emotional and behavioral problems of children and adolescents in Germany--an epidemiological screening. AB - BACKGROUND: Previous studies that were carried out to investigate the prevalence of mental health problems among children and adolescents in Germany are based on regional samples, limited age ranges or used ambiguous case definitions. METHOD: In the Hamburg Health Survey, data on emotional and behavioral problems were collected by a parent and a child questionnaire on the basis of a nationwide representative sample of 1950 families with children and adolescents between the ages of 4 and 18. Besides study-specific items, the Child Behavior Checklist and the Youth Self-Report were used. The symptom and total prevalence rates of clinically significant cases as well as dimensional descriptive statistics and coefficients of agreement between parent- and self-report were calculated. RESULTS: The results indicate that children and adolescents in Germany suffer from a variety of emotional and behavioral problems which require differentiated interpretations depending on their symptom, age and sex. In accordance with the respective case definition, between 10 % and 18% of the children and adolescents were considered to have a clinically relevant score requiring counseling, diagnostics or treatment. The agreement between parent- and self-report scores in the middle range using dimensional statistics and in the lower range for the case definitions. CONCLUSION: The results are discussed in comparison with German and international studies carried out so far. An increase of mental health problems over the past decades cannot be deduced from the data. Procedures which allow the standardization of future investigations are recommended and possibilities to improve current mental health care utilization are discussed. PMID- 15902407 TI - Psychotic symptoms in the general population of England--a comparison of ethnic groups (The EMPIRIC study). AB - BACKGROUND: There is considerable evidence that incidence of schizophrenia and other psychoses varies across ethnic groups in the UK, with particularly high rates for people of African-Caribbean origin. AIMS: The aims of this shady were to estimate in a community-based sample of people from ethnic minorities: 1) the prevalence of psychotic symptoms; and 2) risk factors for reporting psychotic symptoms. METHOD: Face-to-face interviews were carried out with a probabilistic sample of 4281 adults from six ethnic groups living in the UK. Psychotic symptoms were measured using the psychosis screening questionnaire (PSQ). RESULTS: There was a twofold higher rate of reporting psychotic symptoms on the PSQ in Black Caribbean people compared with Whites. Adjustment for demographic factors had little effect on this association. CONCLUSION: Prevalence rates of psychotic symptoms were higher in people from ethnic minorities, but were not consistent with the much higher first contact rates for psychotic disorder reported previously, particularly in Black Caribbeans. PMID- 15902408 TI - Prevalence of mental illness among homeless men in the community--approach to a full census in a southern German university town. AB - AIM: Within the framework of a study of homeless men in the university town of Tubingen in southern Germany, the prevalence of psychiatric disorders and the existing help-seeking behavior, among other things, were to be recorded. METHOD: A total of 151 men belonging to the target group were identified; 91 of them participated in the study. Besides the psychiatric diagnosis, the registered data included psychopathology, cognitive capacity, social functioning and satisfaction with life, as well as social history and case history. RESULTS: Of the probands, 73% were suffering from at least one ongoing psychiatric disorder (diagnosed according to ICD-10 and DSM-IV), primarily alcohol dependence (74%) and drug dependence (34 %), 26% were suffering from an anxiety disorder, 15% from an affective disorder, and 11 % from a disorder of the schizophrenic spectrum. Comorbidity was diagnosed in 67%. Targeted help-seeking behavior concerning the psychiatric or addictive symptoms was extremely rare. Emergency contacts existed with hospitals and general practitioners. DISCUSSION: This study represents the first attempt within Germany and Europe at a full census of homeless men. In international terms, the target region-a small town on the fringes of a conurbation-is a special case. However, compared with studies of help-seeking populations in large cities, virtually no differences were recorded with respect to the prevalence of psychiatric disorders or to help-seeking behavior concerning psychiatric problems and addictions. On the other hand, a substantial number of those concerned evade a resource-intensive, outreach-based study like the present one. Because of German data protection legislation, data on these subjects remain sparse. Concepts aimed at improving the health care system for this target group were found to be dependent in no small measure on the success of the initial contact. PMID- 15902409 TI - Labeling--stereotype--discrimination. An investigation of the stigma process. AB - AIM: Using Link and Phelan's concept of the stigma process, public attitudes towards people with schizophrenia are examined. METHOD: In the spring of 2001, a representative population survey was conducted in Germany (n=5025). A fully structured personal interview was carried out, beginning with the presentation of a case vignette. RESULTS: Labeling as mental illness increased the likelihood that someone suffering from schizophrenia was considered as being unpredictable and dangerous. This, in turn, led to an increase of the preference for social distance. Although much weaker, labeling also had a positive effect on public attitudes insofar as it was associated with a decrease of the tendency to attribute the responsibility for the occurrence of the disorder to the afflicted person. However, this had no significant impact on the desire for social distance. There was no significant association between labeling and the anticipation of poor prognosis. There were some differences between respondents who are familiar with mental illness and those who are not. CONCLUSION: Our findings have some implications for the planning of interventions aimed at reducing stigma and discrimination because of schizophrenia. These interventions should primarily address the stereotypes of unpredictability and dangerousness since they are most likely to have a negative impact on the public's willingness to engage in social relationships with those suffering from this disorder. The interventions should also be tailored according to whether the target population is familiar with mental illness or not. PMID- 15902410 TI - Are self-referrers just the worried well?--A cross-sectional study of self referrers to community psycho-educational Stress and Self-Confidence workshops. AB - BACKGROUND: Reluctance to seek formal help has been seen as a major problem in trying to reduce the prevalence of anxiety and depression. AIMS: The aims of this study were to assess the psychiatric status of those self-referring to psycho educational Stress and Self-Confidence community workshops using a cognitive behavioural therapy (CBT) approach. METHOD: Cross-sectional analysis of 196 people who referred themselves to community workshops was carried out using the Clinical Interview Schedule (CIS-R) psychiatric interview and the Beck Depression Inventory (BDI) and Spielberger Trait Anxiety Inventory (STAI-T) scale self report assessments. RESULTS: Over 70% of all self-referrers had an ICD-10 diagnosis. Those without diagnoses had experienced recurrent significant psychological problems and 29.7% had never consulted their General Practitioner (GP) for anxiety/depression problems. Total CIS-R scores and white ethnic group best predicted previous GP consultation. CONCLUSION: Setting up a self-referral system can enable those with diagnosable psychiatric problems, who may otherwise be reluctant to seek help, to come forward. This may have significant public mental health implications. PMID- 15902411 TI - Factors associated with being a false positive on the General Health Questionnaire. AB - BACKGROUND: The General Health Questionnaire (GHQ) has been used extensively in community and primary care research as an alternative to longer, time-consuming and more expensive assessments of the common mental disorders of depression and anxiety. The sensitivity and specificity of the GHQ compared with longer more detailed assessments is between 70 and 80%. Though satisfactory, this raises the concern about the possibility of bias in relation to longer assessments. We studied socio-demographic factors that were associated with being a false positive on the GHQ in order to investigate any ascertainment bias in relation to more detailed assessments. METHOD: A total of 7,357 consecutive patients aged 16 and over, in five general practices in Cardiff, Bristol and Pontypridd, were invited to complete the 12-item GHQ. Of these, 1,154 patients scored 3 or more, our case definition on the GHQ, and completed a computerised version of the Revised Clinical Interview Schedule (CIS-R) together with a short socio demographic questionnaire. RESULTS: Of the 1,154 subjects who were cases on the GHQ, 344 (30 %) (95% CI 27%-32%) were false positive and were not cases on the CIS-R. After adjustment for the other variables, including GHQ score, false positive subjects were more likely to be employed [odds ratio (OR) 2.7, 95% CI 1.4-5.3], owner-occupiers (OR 1.6, 95% CI 1.0-2.4) and to have a close friend or relative to talk to about personal problems (OR 2.2, 95 % CI 1.4-3.5). CONCLUSION: Our results suggest that in this study there was an ascertainment bias on the GHQ in relation to the CIS-R. Studies that use the GHQ to study the relationship between socio-economic status and common mental disorder could lead to biased estimates of association compared to studies that use the CIS-R. It is likely that the GHQ will lead to a higher estimate of prevalence than the CIS-R in subjects who are better off financially and who have better social support. PMID- 15902412 TI - Person-related predictors of employment outcomes after participation in psychiatric vocational rehabilitation programmes--a systematic review. AB - BACKGROUND: There is increasing recognition of the importance of psychiatric vocational rehabilitation (PVR) programmes in helping individuals with severe mental illnesses to find and secure jobs. However, little is known concerning the factors related to PVR outcomes. OBJECTIVE: This review identifies those person related factors which most strongly influence employment outcomes after participation in PVR programmes. METHOD: Medline, Psychinfo, Pubmed and CINAHL were searched for studies using multivariate analysis of longitudinal data, which analysed employment outcomes after at least 6 months of programme participation, and which analysed at least three predictor domains. RESULTS: Eight studies presented in 16 publications met all the criteria. Better employment outcomes were most strongly related to better work performance measured during PVR participation and to higher work-related self-efficacy. Better social functioning during PVR participation was also significantly related to positive employment outcomes, but this relationship was generally weaker than those of the two aforementioned factors. In most studies, employment outcomes after PVR were not related to past functioning, including work history and diagnosis. Findings on the severity of psychiatric symptoms measured during PVR were mixed. In terms of contribution to outcome, severity of symptoms usually ranked below work performance, when measured concurrently. CONCLUSION: Contrary to previous reviews, this study suggests that the influence of past functioning, including work history, diagnosis and psychiatric history, is outweighed by work performance in PVR. Further prospective and controlled studies are needed to reach more definite conclusions about the individual contributions of person related factors. PMID- 15902413 TI - Occupational abilities and performance scale--reliability-validity assessment factor analysis. AB - BACKGROUND: This article presents a study of the Occupational Abilities and Performance Scale (OAPS), developed for administration to schizophrenic patients. The reliability and validity of the OAPS has been evaluated. METHOD: A total of 174 schizophrenic patients who participated in the Psychosocial and Vocational Rehabilitation Unit (PVRU) of the University Mental Health Research Institute (UMHRI) in Athens were assessed. The OAPS is conducted at entry and after 18 months, when the client has completed training. RESULTS: The results of the reliability analysis showed very good internal consistency, with high split-half reliability as well as test-retest reliability and inter-rater agreement. The scale was also found to have good predictive validity, as well as concurrent validity. Finally, factor analysis with principal components extraction method was performed in order to assess the construct validity of the scale. CONCLUSIONS: The results of factor analysis supported the conclusion of good reliability and validity of the OAPS and revealed the existence of five components, each correlated with a set of the original items. PMID- 15902414 TI - Spontaneous and simultaneous rupture of both quadriceps tendons in a patient with chronic renal failure. A case studied by MRI both preoperatively and postoperatively. AB - Spontaneous bilateral rupture of the quadriceps tendons without a significant history of trauma is an uncommon disease. It is generally associated with chronic metabolic disorders such as chronic renal failure and secondary hyperparathyroidism. Here, we report a case of spontaneous bilateral tendon rupture in a patient on chronic hemodialysis for the past 5 years. We performed a preoperative MRI to confirm the diagnosis; then we repaired the ruptured quadriceps tendons and reviewed the patient both clinically and by performing MRI postoperatively 4 yrs later. The patient attained the full function of the quadriceps tendon. PMID- 15902415 TI - Two minimal incision fasciotomy for chronic exertional compartment syndrome of the lower leg. AB - Chronic exertional compartment syndrome (CECS) of the leg is a pathological condition often related to overuse in subject who engage repetitive physical activities. Fascial release is the mainstay of surgical management. The purpose of this study was to evaluate the results obtained with a double incision decompressive fasciotomy. Eighteen consecutive athletes with a diagnosis of anterior and/or lateral CECS of the leg were operated on with a minimal double incision fascial release after a mean period of 4 months after onset of symptoms. In 11 cases (61%) CECS was bilateral. Surgery was performed without tourniquet and active mobilization was starting immediately. Sports activities were resumed gradually at a mean period of 25 days. The athletes were followed until 2 years. All resumed pre-injury level sports activity. Two patients (18%) of the 11 who underwent to bilateral fasciotomy referred a sensation of leg weakness for an average period of 3 months. The surgical technique presented in this paper seems to be a good mean to treat anterior and lateral leg CECS. The use of tourniquet is deconselled to obtain an accurate intraoperative haemostasis so reducing the risk of post-operative haematoma. PMID- 15902416 TI - Reduced hip bone mineral density is related to physical fitness and leg lean mass in ambulatory individuals with chronic stroke. AB - Following a stroke, the reduced level of physical activity and functional use of the paretic leg may lead to bone loss and muscle atrophy. These factors and the high incidence of falls may contribute to hip fractures in the stroke population. This study was the first to examine total proximal femur bone mineral content (BMC) and bone mineral density (BMD) and their relationship to stroke-specific impairments in ambulatory individuals with chronic stroke (onset >1 year). We utilized dual-energy X-ray absorptiometry (DXA) to acquire proximal femur and total body scans on 58 (23 women) community-dwelling individuals with chronic stroke. We reported total proximal femur BMC (g) and BMD (g/cm2) derived from the proximal femur scans, and lean mass (g) and fat mass (g) for each leg derived from the total body scans. Each subject was evaluated for ambulatory capacity (Six-Minute Walk Test), knee extension strength (hand-held dynamometry), physical fitness [maximal oxygen uptake (VO2max)] and spasticity (Modified Ashworth Scale). Results showed that the paretic leg had significantly lower proximal femur BMD, lean mass and percent lean mass, but higher fat mass than the non paretic leg for both men and women. Proximal femur BMD of the paretic leg was significantly related to ambulatory capacity (r=0.33, P=0.011), muscle strength (r=0.39, P=0.002), physical fitness (r=0.57, P<0.001), but not related to spasticity (r=-0.23, P=0.080). Multiple regression analysis showed that lean mass in the paretic leg was a major predictor (r2=0.371, P<0.001) of the paretic proximal femur BMD. VO2max was a significant predictor of both paretic proximal femur BMD (r2=0.325, P<0.001) and lean mass in the paretic leg (r2=0.700, P<0.001). Further study is required to determine whether increasing physical fitness and lean mass are important to improve hip bone health in chronic stroke. PMID- 15902417 TI - Musculoskeletal manifestations of mild osteogenesis imperfecta in the adult. AB - The musculoskeletal manifestations of mild forms of osteogenesis imperfecta are not well defined in the adult. The aim of this study was to characterize the musculoskeletal manifestations and resulting impairments reported by adults with mild osteogenesis imperfecta. For this task a survey of musculoskeletal symptoms and impairments was hosted on the Osteogenesis Imperfecta Foundation web site for 6 weeks. Survey responses are reported herein. There were 111 unduplicated, adult respondents (78 female). Mean age was 40.8 years. More than one-quarter of 3,410 lifetime fractures occurred in adulthood. Nearly half of respondents reported an established diagnosis of "arthritis" (usually osteoarthritis), and the majority of these reported some degree of impairment attributable to arthritis. Articular pain, stiffness and instability were dominant in the large, weight-bearing joints of the lower extremities. Back pain and scoliosis were common. Of the respondents, 15% required assistance with light physical tasks and personal care. Two-thirds reported joint hyper-mobility, and one-third reported a previous tendon rupture. Complex regional pain syndrome was rare. Respondents reported frequent use of medications known to have potential adverse skeletal effects. In spite of these concerns the majority rated their overall physical health as good or excellent. Adults with mild osteogenesis imperfecta continue to sustain fractures into adulthood, and the majority reports some functional impairment due to musculoskeletal issues. Significant impairment is not rare. PMID- 15902418 TI - Oral contraceptive use in young women is associated with lower bone mineral density than that of controls. AB - Osteoporosis is a skeletal disease affecting 44 million Americans. A primary strategy to prevent osteoporosis is to develop a high peak bone mass in youth. Oral contraceptives (OCs) alter hormones in women and could affect bone mass development. Fifty percent of American women between the ages of 20 and 24 years use OCs. However, the interaction between OCs and skeletal mineralization is poorly understood. Our aim was to compare bone mass [bone mineral density (BMD)] of young women who had a history of OC use, with regularly menstruating controls. We recruited 98 women who were 18 to 25 years of age and had a history of OC use (n=44, 3.4+/-1.9 years of OC use) and controls (n=58). BMD at the hip, whole body, and spine [anterior-posterior (AP) and lateral grams per square centimeter] was measured by dual-energy X-ray absorptiometry (DXA). Physical activity [in metabolic equivalents (METs)] was measured via questionnaire, and grip strength was evaluated with an isometric dynamometer. Groups were similar in body mass index (BMI), fat mass, grip strength, calcium intake and physical activity, but OC users were slightly older than controls (21.3+/-1.9 years vs 20.3+/-1.6 years, P<0.05). In analysis of covariance, controlled for age and BMI, controls had significantly greater BMD than OC users at the AP and lateral spine, femoral neck, greater trochanter, total hip, and whole body (P<0.05). We conclude that, in this cross-sectional analysis, oral contraceptive use by young women may compromise bone health during a time when mineral is still accruing. PMID- 15902419 TI - Toxicant-induced ER-stress and caspase activation in the olfactory mucosa. AB - The potent olfactory toxicant 2,6-dichlorophenyl methylsulphone (2,6-diClPh MeSO(2)) induces rapid cell death and long-term metaplastic changes in the olfactory regions of rodents. The damage is related to a tissue-specific and extensive cytochrome P450 (CYP)-mediated metabolic activation of the compound to reactive intermediates. The aim of the present study was to examine the early, cell-specific changes leading to cell death in the olfactory mucosa of mice exposed to 2,6-diClPh-MeSO(2). We have examined the expression of the ER-specific stress protein GRP78, the presence of secretory glycoproteins, and the cellular activation of the initiator caspase 12 and the downstream effector caspase 3. 2,6 DiClPh-MeSO(2) induced rapid and cell-specific expression of GRP78, and activation of caspases 12 and 3 in the Bowman's glands. No similar early onset changes in the neuroepithelium were observed. Based on these results, we propose that extensive lesions are initiated in the Bowman's glands and that the metabolic activation of 2,6-diClPh-MeSO(2) elicits ER-stress response and subsequent apoptotic signaling at this site. Since most of the Bowman's glands had oncotic morphology, the results suggest that the terminal phase of apoptosis was blocked and that these glands finally succumb to other routes of cell death. PMID- 15902420 TI - Inducing coproporphyria in rat hepatocyte cultures using cyclic AMP and cyclic AMP-releasing agents. AB - Cyclic AMP (c-AMP), added on its own to rat hepatocyte cultures, caused a marked accumulation of coproporphyrin III. The results obtained by comparing the effect of c-AMP to that of exogenous 5-aminolevulinate (ALA), and from adding c-AMP and ALA together, indicated that the coproporphyrinogen III metabolism was blocked, even though no inhibition of the relevant enzyme, coproporphyrinogen oxidase, could be demonstrated. Preferential accumulation of coproporphyrin could also be produced in cultures of rat hepatocytes by agents that raise the cellular levels of cyclic AMP, such as glucagon. The effect of supplementing the culture medium with triiodothyronine (T3) on the response of rat hepatocytes to c-AMP was also investigated. T3, which is known to stimulate mitochondrial respiration, uncoupling O2 consumption from ATP synthesis, produced a c-AMP-like effect when given on its own and potentiated the effect of c-AMP, with an apparent increase in the severity of the metabolic block. It is suggested that an oxidative mechanism may be activated in c-AMP and T3-induced coproporphyria, preferentially involving the mitochondrial compartment, leading to oxidation of porphyrinogen intermediates of haem biosynthesis, especially coproporphyrinogen. Coproporphyin, the fully oxidized aromatic derivative produced, cannot be metabolized and will therefore accumulate. PMID- 15902421 TI - Disposition of low doses of 14C-bisphenol A in male, female, pregnant, fetal, and neonatal rats. AB - Bisphenol A (BPA) is a weak xenestrogen (ADI = 50 microg kg(-1), US EPA) which is mass-produced, with potential for human exposure. To study absorption, distribution, excretion, and metabolism of BPA, BPA labeled with carbon-14 was administered p.o. to male and female Fischer (F344) rats at relatively low doses (20, 100, and 500 microg kg(-1)), and i.v. injected at 100 and 500 microg kg(-1). 14C-BPA (500 microg kg(-1)) was also administered orally to pregnant and lactating rats to examine the transfer of radioactivity to fetuses, neonatal rats, and milk. Radioluminographic determination using phosphor imaging plates was employed to achieve highly sensitive determination of radioactivity. Absorption ratios of radioactivity after three oral doses were high (35-82%); parent 14C-BPA in the circulating blood was quite low, however, suggesting considerable first-pass effect. After an oral dose of 100 microg kg(-1) 14C-BPA, the radioactivity was distributed and eliminated rapidly, but remained in the intestinal contents, liver, and kidney for 72 h. The major metabolite in the plasma and urine was BPA glucuronide, whereas most of the BPA was excreted with the feces as free BPA. A second peak in the time-course of plasma radioactivity suggested enterohepatic recirculation of BPA glucuronide. There was limited distribution of 14C-BPA to the fetus and neonate after oral administration to the dam. Significant radioactivity was not detected in fetuses on gestation days 12 and 15. On day 18, however, radioactivity was detected in the fetal intestine and urinary bladder 24 h after oral dosing of 14C-BPA to the pregnant rats. Part of radioactivity was transferred to neonatal rats from the milk of the treated lactating dam and remained in the intestine of the neonates after 24-h nursing by an untreated dam. PMID- 15902422 TI - Purification and characterization of recombinant human liver prolidase expressed in Saccharomyces cerevisiae. AB - The recombinant human liver prolidase (rh-prolidase, EC 3.4.13.9) from the lysate supernatant of engineering yeast Saccharomyces cerevisiae was purified in two steps employing anion-exchange gradient chromatography (DEAE-Sepharose fast flow) and gel filtration chromatography (Sephacryl S-200 high resolution). The purified recombinant protein furnished a single band with a molecular weight of 56 kD. Intensity scanning of the SDS-PAGE gel revealed that the prolidase accounted for more than 90% of total protein. The optimum pH of the catalytic reaction was 8.0. The enzyme was stimulated by Mn2+, but strongly inhibited by Cu2+ and Zn2+. The rh-prolidase expressed in S. cerevisiae had both dipeptidase and organophosphorus acid anhydrolase activity. It catalyzed the hydrolysis of soman and the dipeptide Gly -Pro. In a detoxification test in vitro, purified rh-prolidase was remarkably efficient at eliminating the toxicity of a lethal dose of soman, with the result that mice survived injection of such a dose. PMID- 15902423 TI - Altered thyroxin and retinoid metabolic response to 2,3,7,8-tetrachlorodibenzo-p dioxin in aryl hydrocarbon receptor-null mice. AB - To determine whether the disruption of thyroid hormone and retinoid homeostasis that occurs after exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) can be mediated by the arylhydrocarbon receptor (AhR), pregnant AhR-heterozygous (AhR+/ ) mice were administered a single oral dose of 10 microg kg(-1) TCDD at gestation day 12.5. Serum and liver were collected on postnatal day 21 from vehicle-treated control or TCDD-treated AhR+/- and AhR-null (AhR-/-) mouse pups. Whereas TCDD exposure resulted in a marked reduction of total thyroxin (TT4) and free T4 (FT4) levels in the serum of AhR+/- mice, TCDD had no effects on AhR-/- mice. Gene expression of UDP-glucuronosyltransferase (UGT)1A6, cytochrome P450 (CYP)1A1, and CYP1A2 in the liver was induced markedly by TCDD in AhR+/- but not AhR-/- mice. Induction of CYP1A1 in response to TCDD was confirmed by immunohistochemical evidence in that CYP1A1 protein was conspicuously localized in the cytoplasm of hepatocytes in the centrilobular region. Levels of retinyl palmitate were greatly reduced in the liver of TCDD-exposed AhR+/- mice, but not in vehicle-treated AhR+/- mice. No effects of TCDD on retinoid levels in the liver were found in AhR /- mice. We conclude that disruption of thyroid hormone and retinoid homeostasis is mediated entirely via AhR. Induction of UGT1A6 is thought to be responsible at least partly for reduced serum thyroid hormone levels in TCDD-exposed mice. PMID- 15902424 TI - Absence of cardiotoxicity of the experimental cytotoxic drug cyclopentenyl cytosine (CPEC) in rats. AB - The experimental anticancer drug cyclopentenyl cytosine (CPEC) was associated with cardiotoxicity in a phase I study. The aim of the present study was twofold; first we investigated whether the observed effects could be reproduced in in vitro and in-vivo rat models. Second, we intended to investigate the underlying mechanism of the possible cardiotoxicity of CPEC. Effects on frequency and contractility were studied on the isolated atria of 18 male Wistar rats. Atria were incubated with 0.1 mmol L(-1) (n = 6) or 1 mmol L(-1) (n = 6) CPEC for 1.5 h and compared with control atria (incubation with buffer solution, n = 6). The cardiac apoptosis-inducing potential was studied in-vivo on 66 rats by 99mTc Annexin V scintigraphy, followed by postmortem determination of radioactivity in tissues, histological confirmation with the TUNEL assay (late-phase apoptosis), and immunohistochemical staining for cleaved caspase 3 and cytochrome C (early phase apoptosis). Serum levels of the necrotic cardiomyopathy marker troponin T were also determined. No effect on heart frequency was found in the isolated atria after CPEC treatment. A trend towards a decrease of contraction force was observed. However, the differences were not statistically significant. 99mTc Annexin V scintigraphy showed no increase in cardiac uptake ratio upon CPEC treatment in the in-vivo rat model, which was confirmed by determination of radioactivity in heart versus blood ratios. At each section a few individual isolated late apoptotic cells (< 5) could be identified by the TUNEL assay in the highest CPEC dose group (90 mg kg(-1)) but not in controls or in rats treated with 60 mg kg(-1) CPEC. Staining for the early apoptosis markers cleaved caspase 3 and cytochrome C did not reveal any significant differences between treated and control rats. Cardiac troponin T levels were not increased after CPEC treatment. CPEC does not affect heart frequency or contraction force in our cardiotoxicity models. Moreover, we did not find an indication of CPEC-induced apoptosis in heart tissue. PMID- 15902425 TI - Weakness in the mechanical properties of the femur of growing female rats exposed to cadmium. AB - This study was aimed at assessing the effect of cadmium (Cd) intoxication on the risk of deformities and fractures of the growing bone on a female rat model of human exposure to this metal. For this purpose, bone mineral density (BMD) and mechanical properties of the proximal and distal ends and diaphysis of the femur were investigated in female Wistar rats exposed to 1, 5, and 50 mg Cd L(-1) in drinking water for 3, 6, 9, and 12 months since weaning. Daily Cd doses received from the drinking water during the treatment period were in the ranges 0.059 0.219, 0.236-1.005, and 2.247-9.649 mg kg(-1) body weight at 1, 5, and 50 mg Cd L(-1), respectively. Biomechanical properties of the femoral proximal and distal ends were evaluated in a compression test and those of the femoral diaphysis in a cutting test with loading perpendicular to the bone longitudinal axis in all tests. Cd dose- and exposure duration-dependently affected the mineralization and mechanical properties of the bone tissue at various locations of the femur. Exposure to 1 mg Cd L(-1) (corresponding to low human exposure) during skeletal development weakened the fracture strength of the femoral neck and of the trabecular bone at the level of the distal end of the femur and affected the elastic properties of the cortical bone at the femoral diaphysis. At the higher levels of Cd treatment, the adverse action generally occurred after shorter exposure than at 1 mg Cd L(-1) and was more seriously advanced. The Cd-induced weakening in the bone biomechanical properties at particular sites of the femur correlated with the decreased bone mineralization. The results indicate that even low exposure to Cd may affect the mineralization and biomechanical properties of growing bone, thus increasing the risk of fractures. PMID- 15902426 TI - Investigation of the role of hyperbaric oxygen therapy in cisplatin-induced nephrotoxicity in rats. AB - Cisplatin (CP) is an effective chemotherapeutic agent used in the treatment of a variety of solid tumours. The most frequently observed side-effect of the use of CP is nephrotoxicity. Recently, evidence has been demonstrated that reactive oxygen species forming in the tubular epithelium play an important role in CP linked nephrotoxicity. The aim of the study was to observe the effect of hyperbaric oxygen (HBO) therapy on CP nephrotoxicity, a subject which has not been studied previously. Wistar rats were treated with CP (a single intraperitoneal (IP) dose of 0.6 mg/100 g) alone and in combination with HBO (60 min every day for seven days at 2.5 x atmospheric pressure). Effects of the treatment on renal function and histology were determined. In analyses at the end of the study it was observed that serum urea, creatinine, and daily urinary protein excretion levels of the CP group were higher than at the start of the study, and that the creatinine clearance level had fallen (P < 0.05). There was no significant difference between the CP+HBO group and HBO group serum urea, creatinine, creatinine clearance, and daily urinary protein excretion levels at the beginning and end of the study (P > 0.05). Histopathological examination showed that the necrosis score in the proximal tubule epithelial cells and average apoptitic cell numbers in the CP group were higher than those in the CP+HBO and HBO groups (P < 0.05). There was no statistical difference between the CP+HBO group and the HBO group in terms of necrosis score in the proximal tubule epithelial cells and the percentage of distal tubules containing hyaline casts in the lumen. In conclusion, in this study it was observed that in experimental study of CP nephrotoxicity the synchronous application of HBO therapy with CP prevents kidney damage. PMID- 15902427 TI - The effect of occlusive and unocclusive exposure to xylene and benzene on skin irritation and molecular responses in hairless rats. AB - Aromatic hydrocarbons readily penetrate the skin on dermal exposure, leading to irritation, inflammation and cytotoxicity. The effects of short-term occlusive and long-term unocclusive dermal exposure to benzene and xylene on the skin irritation response (transepidermal water loss (TEWL), skin moisture content and erythema) and cytokine/chemokine expression (interleukin-1alpha (IL-1alpha), tumor necrosis factor-alpha (TNF-alpha) and monocyte chemoattractant protein-1 (MCP-1)) were investigated in hairless rats. Occlusive dermal exposure was carried out with 230 microL of the chemicals for 1 h using Hill top chambers. In unocclusive dermal exposure, 15 microL of the chemicals were applied to the skin every 2 h, for 8 h a day, for 4 days. The occlusive dermal exposure revealed a clear difference in the TEWL and erythema response of these chemicals (xylene>benzene) whereas unocclusive exposure revealed similar TEWL and erythema scores for both benzene and xylene. The expression of IL-1alpha was elevated 2.5- and 3.8-fold in response to occlusive and unocclusive exposure, respectively, vs control (P<0.01) for both the chemicals (benzene and xylene). Similarly, TNF alpha levels were elevated about 2.4- and 6.0-fold as a result of occlusive and unocclusive exposure, respectively, vs control (P<0.01). These results show that unocclusive exposure induced significantly higher TNF-alpha expression than occlusive exposure (P<0.05). The MCP-1 expression in blood was slightly elevated compared with the control group, but this increase was not statistically significant (P>0.05). Similarly, MCP levels in skin were increased approximately 1.7- and 1.8-fold by occlusive and unocclusive exposure, respectively, compared with the control group (P<0.05). Our study demonstrates that the skin irritation profiles of benzene and xylene are similar and unocclusive long-term exposure to small amounts of these chemicals can induce more skin irritation and cytokine response than occlusive exposure. PMID- 15902428 TI - Reduction of myocardial infarct size by SM-198110, a novel Na+/H+ exchange inhibitor, in rabbits. AB - The effects of 3-[2-({[amino(imino)methyl]amino}carbonyl)-4-chloro-1H-indol-1-yl] 1-propanesulphonic acid monohydrate (SM-198110), a novel potent Na+/H+ exchange inhibitor, and cariporide (Hoe642), another Na+/H+ exchange inhibitor, were studied in a myocardial ischaemia and reperfusion injury model. Anaesthetized rabbits were subjected to occlusion of the coronary artery for 30 min followed by reperfusion for 5 h. SM-198110 or cariporide was administered before ischaemia and before reperfusion. We also assessed the anti-necrotic effect of SM-198110 when given before reperfusion, both alone and together with glibenclamide, a K(ATP) channel blocker, 5-hydroxydecanoate (5-HD), a mitochondrial K(ATP) channel selective blocker and 8-(p-sulphophenyl)-theophylline (8-SPT), an adenosine receptor blocker. The infarct size was reduced dose-dependently by i.v. administration of SM-198110 before ischaemia, with a significant reduction in serum creatine phosphokinase activity. Infarct sizes, normalized to the size of the area-at-risk (means+/-SE) were: vehicle 56.6+/-3.7%; low-dose SM-198110 39.2+/-6.3%; mid-dose 32.8+/-7.4% (P < 0.05); high-dose 22.1+/-6.7% (P < 0.01). This anti-necrotic effect of SM-198110 was achieved without significant haemodynamic changes. Cariporide given before ischaemia also reduced infarct size significantly and dose-dependently. SM-198110 administered before reperfusion also resulted in a dose-dependent reduction in the infarct size. Infarct sizes were: vehicle 56.6+/-3.7%; low-dose SM-198110 44.5+/-5.7%; mid-dose 36.3+/-6.6% (P < 0.01); high-dose 34.7+/-3.8% (P < 0.01). In contrast, cariporide given before reperfusion did not reduce infarct sizes significantly. The anti-necrotic effect of SM-198110 was observed even when given 10 min after the beginning of reperfusion. Glibenclamide and 5-HD abolished the anti-necrotic effect of treatment before reperfusion with SM-198110. However, the co-administration of 8 SPT with SM-198110 did not affect infarct size. These results suggest that, in addition to Na+/H+ exchange inhibition, mitochondrial and/or sarcolemmal K(ATP) channels contribute to the anti-necrotic effect of SM-198110 when the latter is given before reperfusion. PMID- 15902429 TI - Emerging roles of TRPM6/TRPM7 channel kinase signal transduction complexes. AB - Investigations into Drosophila mutants with impaired vision due to mutations in the transient receptor potential gene (trp) initiated a systematic search for TRP homologs in other species, finally leading to the discovery of a whole new family of plasma membrane cation channels involved in multiple physiological processes. Among the recently discovered TRP cation channels two homologous proteins, TRPM6 and TRPM7, display unique domain compositions and biophysical properties. These remarkable genes are vital for Mg(2+) homeostasis in vertebrates and, if disrupted, lead to cell death or human disease. PMID- 15902431 TI - Complications of lumbar puncture in a child treated for leukaemia. AB - Lumbar puncture may lead to neurological complications. These include intracranial hypotension, cervical epidural haematomas, and cranial and lumbar subdural haematomas. MRI is the modality of choice to diagnose these complications. This report documents MRI findings of such complications in a child treated for leukaemia. PMID- 15902430 TI - Receptor-operated cation channels formed by TRPC4 and TRPC5. AB - TRPC4 and TRPC5 form cation channels that contribute to phospholipase C-dependent Ca(2+) entry following stimulation of G-protein-coupled receptors or receptor tyrosine kinases. Surprisingly, in different studies, TRPC4 and TRPC5 have been shown to form either store-operated channels with a relatively high Ca(2+) permeability, or nonselective cation channels activated independently of store depletion. In this review, we summarize and discuss data on the regulation and permeability properties of TRPC4 and TRPC5, and data on native channels that might be composed of these isoforms. PMID- 15902432 TI - Spontaneous development of bilateral subdural hematomas in an infant with benign infantile hydrocephalus: color Doppler assessment of vessels traversing extra axial spaces. AB - We present an infant with macrocrania, who initially demonstrated prominent extra axial fluid collections on sonography of the brain, compatible with benign infantile hydrocephalus (BIH). Because of increasing macrocrania, a follow-up sonogram of the brain was performed; it revealed progressive enlargement of the extra-axial spaces, which now had echogenic debris. Color Doppler US showed bridging veins traversing these extra-axial spaces, so it was initially thought that these spaces were subarachnoid in nature (positive cortical vein sign). However, an arachnoid membrane was identified superior to the cortex, and there was compression of true cortical vessels beneath this dural membrane. An MRI of the brain showed the extra-axial spaces to represent bilateral subdural hematomas. The pathogenesis of spontaneous development of the subdural hematomas, in the setting of BIH, is discussed. We also emphasize that visualizing traversing bridging veins through extra-axial spaces does not necessarily imply that these spaces are subarachnoid in origin. PMID- 15902433 TI - Moderate sedation for MRI in young children with autism. AB - Autism is a pervasive neurodevelopmental disorder. Because of the deficits associated with the condition, sedation of children with autism has been considered more challenging than sedation of other children. OBJECTIVE: To test this hypothesis, we compared children with autism against clinical controls to determine differences in requirements for moderate sedation for MRI. MATERIALS AND METHODS: Children ages 18-36 months with autism (group 1, n = 41) and children with no autistic behavior (group 2, n = 42) were sedated with a combination of pentobarbital and fentanyl per sedation service protocol. The sedation nurse was consistent for all patients, and all were sedated to achieve a Modified Ramsay Score of 4. Demographics and doses of sedatives were recorded and compared. RESULTS: There were no sedation failures in either group. Children in group 1 (autism) were significantly older than group 2 (32.02+/-3.6 months vs 28.16+/-6.7 months) and weighed significantly more (14.87+/-2.1 kg vs 13.42+/-2.2 kg). When compared on a per-kilogram basis, however, group 1 had a significantly lower fentanyl requirement than group 2 (1.25+/-0.55 mcg/kg vs 1.57+/-0.81 mcg/kg), but no significant difference was found in pentobarbital dosing between groups 1 and 2, respectively (4.92+/-0.92 mg/kg vs 5.21+/-1.6 mg/kg). CONCLUSION: Autistic children in this age range are not more difficult to sedate and do not require higher doses of sedative agents for noninvasive imaging studies. PMID- 15902434 TI - Prenatal diagnosis of horseshoe lung: contribution of MRI. AB - Horseshoe lung is a very rare pulmonary anomaly that is characterized by an isthmus of lung parenchyma bridging the right and left lungs and extending through the mediastinum. We report on the prenatal diagnosis of such a malformation in a 33-week-gestation fetus. The diagnosis was initially suspected on antenatal ultrasonography performed at 33 weeks and confirmed by fetal MRI and subsequent pathological examination after termination of pregnancy. To our knowledge, this is the first reported case of antenatal diagnosis of horseshoe lung. PMID- 15902435 TI - Mice lacking the IFN-gamma receptor or fyn develop severe experimental autoimmune uveoretinitis characterized by different immune responses. AB - Endogenous interferon (IFN)-gamma negatively regulates experimental autoimmune uveoretinitis (EAU), a Th1-mediated disease. Although it is well known that IFN gamma exerts its effects by binding to the IFN-gamma receptor (IFN-gammaR), the role that IFN-gammaR plays in the development of EAU has not been investigated. Fyn has been reported to inhibit Th2 differentiation. We aimed to investigate how endogenous IFN-gammaR and fyn, which influence Th1/Th2 differentiation, participate in the development of EAU. Sex-matched 6- to 10-week-old C57BL/6 wild type (WT), IFN-gammaR knockout (GRKO) and fyn knockout (fyn KO) mice were compared. Mice were immunized subcutaneously with human interphotoreceptor retinoid-binding protein peptide 1-20 emulsified in Freund's complete adjuvant together with an intraperitoneal injection of Bordetella pertussis toxin. Three weeks later, mice were sacrificed, and their eyes and spleens were harvested for histopathologic analyses and examination of cellular immune responses, respectively. Cellular immune responses were evaluated by measuring the proliferative responses and cytokine production [interleukin (IL)-4, IL-5, IL-6, IL-13, IFN-gamma and tumor necrosis factor (TNF)-alpha] of splenocytes. The incidence of EAU was 40.0% in WT mice, 59.3% in GRKO mice and 78.6% in fyn KO mice. The average EAU score was 0.294 in WT mice, 0.917 in GRKO mice and 1.063 in fyn KO mice. Upon EAU induction, significant infiltration of eosinophils into the eyes was observed in GRKO and fyn KO mice compared to WT mice. Splenocytes from GRKO mice proliferated against the antigen and a mitogen more vigorously than those from WT and fyn KO mice. Stimulation of splenocytes with the antigen induced a higher production of IL-4, IL-6, IL-13 and IFN-gamma in GRKO mice compared to WT and fyn KO mice. In contrast, IL-5 and TNF-alpha were most abundantly produced by splenocytes from fyn KO mice compared to WT and GRKO mice. The incidence and mean severity of EAU were significantly higher in GRKO and fyn KO mice than in WT mice, suggesting that endogenous IFN-gammaR and fyn negatively regulate the development of EAU. The different cytokine production patterns by the GRKO and fyn KO mice indicate that the negative regulatory mechanism mediated by IFN-gammaR and fyn may differ. PMID- 15902436 TI - Identification and characterization of the rat homologue of LAIR-1. AB - Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) is a cell-surface molecule that functions as an inhibitory receptor on various immune cells in both humans and mice. We have cloned a LAIR-1 homologue from the rat that we have named rat LAIR-1. The LAIR-1 gene maps to rat chromosome 1q12 in a region showing conserved synteny with human chromosome 19q13.4 and mouse chromosome 7, where the leukocyte receptor cluster is located. Rat LAIR-1 shows 40 and 71% protein sequence identity with human LAIR-1 and mouse LAIR-1, respectively, has a single Ig-like domain and contains two immunoreceptor tyrosine-based inhibitory motif like sequences in its cytoplasmic tail. Soluble rat LAIR-1 fusion proteins bind to the same adherent cell lines as human LAIR-1 and mouse LAIR-1, indicating that a putative ligand for all the LAIR-1 molecules is expressed on these cells. Furthermore, we show that rat and mouse LAIR-1 bind the same molecule expressed on human HT29 cells. Since many autoimmune diseases are studied in rat models, identification of rat LAIR-1 allows for in vivo studies on the function of LAIR molecules in these systems. PMID- 15902437 TI - To use MIBI or not to use MIBI? That is the question when assessing tumour cells. AB - 99mTc-sestamibi (MIBI) is a well-known tumour imaging agent. Its retention within tumour cell mitochondria is related to perfusion and to the magnitude of the electrical gradient, reflecting cell viability. Several internal cell factors modulate this uptake; for example, multidrug resistance membrane proteins (Pgp and MRP1) and anti-apoptotic BCl-2 protein of the outer mitochondrial membrane can limit retention of MIBI. At the early stage of cell apoptosis, the electrical driving forces of MIBI uptake are impaired, and influx and accumulation are reduced. It seems clear that MIBI can be used before treatment to detect drug resistance, assess anti-apoptotic status and predict treatment efficacy. Although it has been suggested that MIBI might be used to monitor tumour response to treatment, MIBI is unable to differentiate tumours with ongoing apoptosis from those developing drug resistance. PMID- 15902438 TI - Influence of collimator choice and simulated clinical conditions on 123I-MIBG heart/mediastinum ratios: a phantom study. AB - PURPOSE: (123)I presents imaging problems owing to high-energy photon emission. We investigated the influence of collimators on (123)I-MIBG heart/mediastinum ratios (H/M ratios). Secondly, we assessed the influence on H/M ratios of different activity concentrations, simulating clinical conditions. Thirdly, the value of scatter correction was assessed. METHODS: The AGATE cardiac phantom was filled with (123)I in three sequential conditions: A, heart and mediastinal activity; B, adding lung activity; and C, adding liver activity (protocol I). In protocol II, myocardium and liver were filled with different activities ranging from low to high. For each condition, static anterior planar and single-photon emission computed tomography studies were acquired on a Siemens e.cam (SI) and a General Electric Millennium VG (GE) system, using low-energy high-resolution and medium-energy (ME) collimators for protocol I and only ME collimators for protocol II . For the SI camera, a triple energy window (TEW) scatter correction was applied. RESULTS: Planar H/M ratios were influenced by scatter and septal penetration from increasing amounts of liver activity. These effects were less pronounced for ME collimators. Although the TEW scatter correction increased ratios overall, TEW correction did not improve the relative differences between the ratios. TEW correction therefore does not add any benefit to obtain an accurate reflection of myocardial activity concentrations. CONCLUSION: For straightforward implementation of semi-quantitative (123)I-MIBG myocardial studies, we recommend the use of ME collimators without scatter correction. PMID- 15902439 TI - Validation of brain tumour imaging with p-[123I]iodo-L-phenylalanine and SPECT. AB - PURPOSE: The aims of this prospective study were to validate single-photon emission computed tomography (SPECT) with p-[(123)I]iodo-L-phenylalanine (IPA) in brain tumours and to evaluate its potential for the characterisation of indeterminate brain lesions. METHODS: In 45 patients with indeterminate brain lesions or suspected progression of glioma, amino acid uptake was studied using IPA-SPECT and compared with the final diagnosis established by biopsy or serial imaging. After image fusion of IPA-SPECT and magnetic resonance imaging, the presence of tumour was visually determined by two independent observers. IPA uptake was quantified as the ratio between maximum uptake in the suspicious lesion and mean uptake in unaffected brain. RESULTS: Primary brain tumours were present in 35 cases (12 low-grade and 23 high-grade gliomas). Non-neoplastic brain lesions were confirmed in seven cases (three dysplasias, three inflammatory lesions, one lesion after effective therapy). Visual analysis showed a high concordance between the two observers (kappa=0.90, p<0.001), with sensitivity and specificity of 86% and 100% for the discrimination of primary brain tumours and non-neoplastic lesions. At 30 min p.i., IPA uptake in primary brain tumours was higher than that in non-neoplastic lesions (1.70+/-0.36 vs 1.14+/-0.18, p<0.05). Brain metastases showed no increased uptake (1.13+/-0.22, n=3). The persistent retention of IPA in low-grade gliomas without disruption of the blood-brain barrier was visualised up to 24 h p.i. Low-grade and high-grade gliomas showed equivalent IPA uptake (1.72+/-0.37 vs 1.67+/-0.36 at 30 min, p=0.745). CONCLUSION: IPA shows long and specific retention in gliomas. IPA is a promising and safe radiopharmaceutical for the visualisation of gliomas and the characterisation of indeterminate brain lesions. PMID- 15902440 TI - Comparison of IgG and F(ab')2 fragments of bispecific anti-RCCxanti-DTIn-1 antibody for pretargeting purposes. AB - PURPOSE: An effective pretargeting strategy was developed for renal cell carcinoma (RCC) based on a biologically produced bispecific monoclonal antibody: anti-RCCxanti-DTPA(In) (bsMAb: G250xDTIn-1). Tumour uptake of a (111)In-labelled bivalent peptide after pretargeting with bsMAb G250xDTIn-1 was relatively high compared with that in other pretargeting systems using chemically coupled F(ab')(2) fragments. Here, we investigated the effect of the bsMAb form in the pretargeting strategy. METHODS: To determine the optimal interval between the administration of each of the bsMAb forms and the (111)In-labelled bivalent peptide, the biodistribution of the radioiodinated bsMAb forms was studied in athymic mice with subcutaneous SK-RC-1 RCC tumours. Since tumour targeting of the radiolabelled peptide depends on the bsMAb form and dose, a bsMAb dose escalation study was carried out for both bsMAb forms. Under optimised conditions, the biodistribution of the (111)In label in mice with pretargeted RCC was determined from 4 h up to 7 days p.i. RESULTS: The optimal interval between the two administrations was 72 h for the bsMAb IgG and 4 h for the bsMAb F(ab')(2). The optimal bsMAb dose for intact IgG was 67 pmol and the optimal bsMAb F(ab')(2) dose was 200 pmol. Targeting of the pretargeted RCC with 4 pmol (111)In-labelled bivalent peptide revealed high tumour uptake with both bsMAb forms. CONCLUSION: With the pretargeting strategy, using either bsMAb IgG or bsMAb F(ab')(2), very efficient peptide targeting of the tumour was obtained. Uptake and retention of the radiolabel in the tumour with the pretargeting approach are not affected by the bsMAb form used. PMID- 15902441 TI - Improving the urban stream restoration effort: identifying critical form and processes relationships. AB - Stream restoration projects are often based on morphological form or stream type and, as a result, there needs to be a clear tie established between form and function of the stream. An examination of the literature identifies numerous relationships in naturally forming streams that link morphologic form and stream processes. Urban stream restoration designs often work around infrastructure and incorporate bank stabilization and grade control structures. Because of these imposed constraints and highly altered hydrologic and sediment discharge regimens, the design of urban channel projects is rather unclear. In this paper, we examine the state of the art in relationships between form and processes, the strengths and weaknesses of these existing relationships, and the current lack of understanding in applying these relationships in the urban environment. In particular, we identify relationships that are critical to urban stream restoration projects and provide recommendations for future research into how this information can be used to improve urban stream restoration design. It is also suggested that improving the success of urban restoration projects requires further investigation into incorporating process-based methodologies, which can potentially reduce ambiguity in the design and the necessity of using an abundant amount of in-stream structures. PMID- 15902442 TI - Forest and farmland conservation effects of Oregon's (USA) land-use planning program. AB - Oregon's land-use planning program is often cited as an exemplary approach to forest and farmland conservation, but analyses of its effectiveness are limited. This article examines Oregon's land-use planning program using detailed spatial data describing building densities in western Oregon. An empirical model describes changes in building densities on forest and agricultural lands from 1974 to 1994, as a function of a gravity index of land's commuting distance to cities of various sizes, topographic characteristics, and zoning adopted under Oregon's land-use planning program. The effectiveness of Oregon's land-use planning program is evaluated based on the statistical significance of zoning variables and by computing estimated areas of forest and agricultural lands falling into undeveloped, low-density developed, and developed building density categories, with and without land-use zoning in effect. Results suggest that Oregon's land-use planning program has provided a measurable degree of protection to forest and agricultural lands since its implementation. PMID- 15902443 TI - Influence of catchment-scale military land use on stream physical and organic matter variables in small southeastern plains catchments (USA). AB - We conducted a 3-year study designed to examine the relationship between disturbance from military land use and stream physical and organic matter variables within 12 small (<5.5 km2) Southeastern Plains catchments at the Fort Benning Military Installation, Georgia, USA. Primary land-use categories were based on percentages of bare ground and road cover and nonforested land (grasslands, sparse vegetation, shrublands, fields) in catchments and natural catchments features, including soils (% sandy soils) and catchment size (area). We quantified stream flashiness (determined by slope of recession limbs of storm hydrographs), streambed instability (measured by relative changes in bed height over time), organic matter storage [coarse wood debris (CWD) relative abundance, benthic particulate organic matter (BPOM)] and stream-water dissolved organic carbon concentration (DOC). Stream flashiness was positively correlated with average storm magnitude and percent of the catchment with sandy soil, whereas streambed instability was related to percent of the catchment containing nonforested (disturbed) land. The proportions of in-stream CWD and sediment BPOM, and stream-water DOC were negatively related to the percent of bare ground and road cover in catchments. Collectively, our results suggest that the amount of catchment disturbance causing denuded vegetation and exposed, mobile soil is (1) a key terrestrial influence on stream geomorphology and hydrology and (2) a greater determinant of in-stream organic matter conditions than is natural geomorphic or topographic variation (catchment size, soil type) in these systems. PMID- 15902444 TI - A quantitative tool for assessing the integrity of southern coastal California streams. AB - We developed a benthic macro-invertebrate index of biological integrity (B-IBI) for the semi-arid and populous southern California coastal region. Potential reference sites were screened from a pool of 275 sites, first with quantitative GIS landscape analysis at several spatial scales and then with local condition assessments (in-stream and riparian) that quantified stressors acting on study reaches. We screened 61 candidate metrics for inclusion in the B-IBI based on three criteria: sufficient range for scoring, responsiveness to watershed and reach-scale disturbance gradients, and minimal correlation with other responsive metrics. Final metrics included: percent collector-gatherer + collector-filterer individuals, percent non-insect taxa, percent tolerant taxa, Coleoptera richness, predator richness, percent intolerant individuals, and EPT richness. Three metrics had lower scores in chaparral reference sites than in mountain reference sites and were scored on separate scales in the B-IBI. Metrics were scored and assembled into a composite B-IBI, which was then divided into five roughly equal condition categories. PCA analysis was used to demonstrate that the B-IBI was sensitive to composite stressor gradients; we also confirmed that the B-IBI scores were not correlated with elevation, season, or watershed area. Application of the B-IBI to an independent validation dataset (69 sites) produced results congruent with the development dataset and a separate repeatability study at four sites in the region confirmed that the B-IBI scoring is precise. The SoCal B-IBI is an effective tool with strong performance characteristics and provides a practical means of evaluating biotic condition of streams in southern coastal California. PMID- 15902445 TI - Monitoring environmental changes in the Mediterranean coastal landscape: the case of Cukurova, Turkey. AB - This paper describes a remote sensing approach used to monitor temporal land use/cover (LULC) changes in Cukurova, an extensive coastal plain in the southeast Mediterranean coast of Turkey. The area has varied terrain ranging from low-lying alluvial deposits to rocky hills and mountains characterized by limestone outcrops. The ecological and economic importance of the area can be attributed to the existence of important coastal ecosystems (e.g., wetlands and sand dunes) and a wide range of industries located along the eastern coast. Temporal changes in the coastal landscape between 1984 and 2000 were evaluated using digital interpretation of remotely sensed satellite data. Pairwise comparison methods were used to quantify changes from 1984 to 1993 and 1993 to 2000 using multitemporal Landsat TM and ETM+ images, acquired in 1984, 1993, and 2000, respectively. Total change area was 2448 ha from 1984 to 1993 and increased more than twofold, to 6072 ha from 1993 to 2000. Change trends were determined using the information provided from individual change detection outputs of different periods. The most prominent changes were estimated to have occurred in agriculture, urban, and natural vegetation cover. Agriculture has increasingly grown over marginal areas, whereas urban development occurred at the expense of prime croplands across both time steps. PMID- 15902446 TI - Assessing ecological integrity of Ozark rivers to determine suitability for protective status. AB - Preservation of extraordinary natural resources, protection of water quality, and restoration of impaired waters require a strategy to identify and protect least disturbed streams and rivers. We applied two objective, quantitative methods to determine stream ecological integrity of headwater reaches of 10 Ozark rivers, 5 with Wild and Scenic River federal protective status. Thirty-four variables representing macroinvertebrate and fish assemblage characteristics, in-stream habitat, riparian vegetation, water quality, and watershed attributes were quantified for each river and analyzed using two multivariate approaches. The first approach, cluster and discriminant analyses, identified two groups of river with only one variable (% forested watershed) reliably distinguishing groups. Our second approach employed ordinal scaling to compare variables for each river to conceptually ideal conditions that were developed as a composite of optimal attributes among the 10 rivers. The composite distance of each river from ideal was then calculated using a unidimensional ranking technique. Two rivers without Wild and Scenic River designation ranked highest relative to ideal (highest ecological integrity), and two others, also without designation, ranked most distant from ideal (lowest ecological integrity). Fish density, number of intolerant fish species, and invertebrate density were influential biotic variables for scaling. Contributing physical variables included riparian forest cover, water nitrate concentration, water turbidity, percentage of forested watershed, percentage of private land ownership, and road density. These methods provide a framework for refinement and application in other regions to facilitate the process of establishing least-disturbed reference conditions and identifying rivers for protection and restoration. PMID- 15902447 TI - Corporate environmental policy statements in mainland China: to what extent do they conform to ISO 14000 documentation? AB - For decades, industry has been the main source of pollution in China. Determined to make changes, the mainland Chinese authorities have decided to promote mechanisms that incorporate environmental concerns into the internal management of enterprises. This is manifested in the rapid adoption of the ISO14000 standards, including a significant increase in ISO14001 registrations in China. Thus, this study examined the environmental policy statements of 106 certified facilities in mainland China against a strict interpretation of the mandatory requirements of the ISO 14001:1996 standard and the nonmandatory ISO14004 requirements in order to shed some light on the implementation of environmental management systems in an emerging economic giant. It was decided to analyze the environmental policy statement because such a statement is a core element in the ISO system of environmental management of each facility and there are relatively clear and specific requirements on what an environmental policy statement shall include. An analysis of the contents of the environmental policy statements shows that conformance to the relevant requirements of both the mandatory ISO14001 standard and the nonmandatory ISO14004 standard is far from impressive and that the facilities in our sample seldom went beyond the minimum requirements. By using ISO14001 and ISO14004 conformance scores as the dependent variables, we found that conformity to ISO14001 and overall conformance to ISO14000 series can be explained to some extent by the degree of top management commitment, the experience with informal environmental management systems, and the form of ownership of the facilities. PMID- 15902448 TI - Testing the use of a land cover map for habitat ranking in boreal forests. AB - Habitat loss and modification is one of the major threats to biodiversity and the preservation of conservation values. We use the term ''conservation value'' to mean the benefit of nature or habitats for species. The importance of identifying and preserving conservation values has increased with the decline in biodiversity and the adoption of more stringent environmental legislation. In this study, conservation values were considered in the context of land-use planning and the rapidly increasing demand for more accurate methods of predicting and identifying these values. We used a k-nearest neighbor interpreted satellite (Landsat TM) image classified in 61 classes to assess sites with potential conservation values at the regional and landscape planning scale. Classification was made at the National Land Survey of Finland for main tree species, timber volume, land-use type, and soil on the basis of spectral reflectance in satellite image together with broad numerical reference data. We used the number and rarity of vascular plant species observed in the field as indicators for potential conservation values. We assumed that significant differences in the species richness, rarity, or composition of flora among the classes interpreted in the satellite image would also mean a difference in conservation values among these classes. We found significant differences in species richness among the original satellite image classes. Many of the classes examined could be distinguished by the number of plant species. Species composition also differed correspondingly. Rare species were most abundant in old spruce forests (>200 m3/ha), raising the position of such forests in the ranking of categories according to conservation values. The original satellite image classification was correct for 70% of the sites studied. We concluded that interpreted satellite data can serve as a useful source for evaluating habitat categories on the basis of plant species richness and rarity. Recategorization of original satellite image classification into such new conservation value categories is challenging because of the variation in species composition among the new categories. However, it does not represent a major problem for the purposes of early-stage land-use planning. Benefits of interpreted satellite image recategorization as a rapid conservation value assessment tool for land-use planners would be great. PMID- 15902449 TI - Using satellite fire detection to calibrate components of the fire weather index system in Malaysia and Indonesia. AB - Vegetation fires have become an increasing problem in tropical environments as a consequence of socioeconomic pressures and subsequent land-use change. In response, fire management systems are being developed. This study set out to determine the relationships between two aspects of the fire problems in western Indonesia and Malaysia, and two components of the Canadian Forest Fire Weather Index System. The study resulted in a new method for calibrating components of fire danger rating systems based on satellite fire detection (hotspot) data. Once the climate was accounted for, a problematic number of fires were related to high levels of the Fine Fuel Moisture Code. The relationship between climate, Fine Fuel Moisture Code, and hotspot occurrence was used to calibrate Fire Occurrence Potential classes where low accounted for 3% of the fires from 1994 to 2000, moderate accounted for 25%, high 26%, and extreme 38%. Further problems arise when there are large clusters of fires burning that may consume valuable land or produce local smoke pollution. Once the climate was taken into account, the hotspot load (number and size of clusters of hotspots) was related to the Fire Weather Index. The relationship between climate, Fire Weather Index, and hotspot load was used to calibrate Fire Load Potential classes. Low Fire Load Potential conditions (75% of an average year) corresponded with 24% of the hotspot clusters, which had an average size of 30% of the largest cluster. In contrast, extreme Fire Load Potential conditions (1% of an average year) corresponded with 30% of the hotspot clusters, which had an average size of 58% of the maximum. Both Fire Occurrence Potential and Fire Load Potential calibrations were successfully validated with data from 2001. This study showed that when ground measurements are not available, fire statistics derived from satellite fire detection archives can be reliably used for calibration. More importantly, as a result of this work, Malaysia and Indonesia have two new sources of information to initiate fire prevention and suppression activities. PMID- 15902450 TI - Possible input of nitrogen of visitors' origin on a protected peatland. AB - The Ozegahara peatland, in the Nikko-Oze National Park in Japan, is ecologically significant because of its oligotrophic environment; it is one of the most strictly preserved areas in the country. The isotope ratio of nitrogen (15N/14N) and carbon (13C/12C) and C/N ratio of peat moss (Sphagnum spp.) and sundew (Drosera rotundifolia) in the peatland were analyzed. The correlation of the isotope ratio with some parameters (sundew population density, number of trapped insects, water level, bog myrtle coverage, and visitor density) was investigated. The nitrogen isotope ratio of sundew showed the most significant covariation with visitor density, where sundew from lunch areas or along busy walkways showed a higher nitrogen isotope ratio. The nitrogen isotope ratio of peat moss covaried, not only with route traffic frequency but also with water level, bog myrtle coverage, and number of trapped insects by sundew, indicating that factors other than the visitor level influence the local nitrogen cycle. This study suggests that the nitrogen imported into the peatland by visitors is a principal factor to be monitored for the maintenance of the natural environment. PMID- 15902451 TI - Household demand for waste recycling services. AB - Municipalities everywhere are coping with increasing amounts of solid waste and need urgently to formulate efficient and sustainable solutions to the problem. This study examines the use of economic incentives in municipal waste management. Specifically, we address the issue of recycling, if and when this waste management option is-on social welfare grounds-a preferred solution.A number of studies have recently assessed the monetary value of the externalities of alternative solid waste management options. In the present context, these subsidies could be interpreted as the implicit value of the benefits from reducing environmental externalities associated with landfilling as perceived by local government authorities. We surmise that the difference between mean households' willingness to pay (WTP) for recycling services, via the purchase of a subsidized waste disposal facility, and the above (proxy) value of externalities reflects the difference between private and public perception regarding the negative externality associated with landfilling. We believe that this information is useful in determining the level of subsidization needed (if at all) to sustain any recycling program.The study is unique in the sense that its conclusions are based on revealed household behavior when faced with increased disposal costs, as well as information on WTP responses in hypothetical but related (and, therefore, familiar) scenarios. The article also explores the influence of the subsidization schemes on recycling rates. It was found that with low levels of effort needed to participate in a curbside recycling program, households' participation rates are mainly influenced by economic variables and age, and households are willing to pay a higher price for the recycling scheme. When the required effort level is relatively high, however, households would pay a lower price, and the rate is influenced mainly by their environmental commitment and by economic considerations. We found that in both cases a subsidy would be required in order to achieve an efficient level of recycling. The median price that households are willing to pay for recycling devices is found to be about NIS 370 (New Israeli Shekel, approximately 90 dollars). PMID- 15902452 TI - Influence of targets and assessment region size on perceived conservation priorities. AB - We used an existing conservation opportunity area (OA) data layer for four contiguous ecological subsections within the Ozark Highlands to quantitatively evaluate the influence of conservation targets and assessment region size on conservation priorities. OAs are natural and seminatural land-cover patches that are away from roads and away from patch edges. To evaluate the influence of targets, we assigned a priority score to each OA polygon for each of five different conservation targets, including land-cover patch size, landform representation, target vertebrate richness, target breeding bird richness, and target land cover. The top-scoring OAs for each target were added to an OA selection set for that target until 50% of the study area was chosen. These five OA selection sets were overlain to quantify overlap in priorities. Only 1.6% of the study area, or 2.1% of all OA polygons, was selected by all five targets. To evaluate the influence of assessment region size, we compared results of priority ranking of OAs relative to the entire study area against a merged set of priority rankings established separately relative to each of the four subsections within the study area. When high-priority OAs were added until 25% of the region was within the selection set for each of the five targets, the sets based on the whole study area versus each subsection evaluated separately overlapped from 45.4% to 81.9%. Thus, perceived priorities of conservation assessments are strongly influenced both by the targets that are evaluated and by the size of the assessment region. PMID- 15902453 TI - Theory into practice: implementing ecosystem management objectives in the USDA Forest Service. AB - In the United States and around the world, scientists and practitioners have debated the definition and merits of ecosystem management as a new approach to natural resource management. While these debates continue, a growing number of organizations formally have adopted ecosystem management. However, adoption does not necessarily lead to successful implementation, and theories are not always put into practice. In this article, we examine how a leading natural resource agency, the United States Department of Agriculture Forest Service, has translated ecosystem management theory into concrete policy objectives and how successfully these objectives are perceived to be implemented throughout the national forest system. Through document analysis, interviews, and survey responses from 345 Forest Service managers (district rangers, forest supervisors, and regional foresters), we find that the agency has incorporated numerous ecosystem management components into its objectives. Agency managers perceive that the greatest attainment of such objectives is related to collaborative stewardship and integration of scientific information, areas in which the organization has considerable prior experience. The objectives perceived to be least attained are adaptive management and integration of social and economic information, areas requiring substantial new resources and a knowledge base not traditionally emphasized by natural resource managers. Overall, success in implementing ecosystem management objectives is linked to committed forest managers. PMID- 15902454 TI - The appropriateness of snowmobiling in national parks: an investigation of the meanings of snowmobiling experiences in yellowstone national park. AB - The debate over snowmobiling and other types of motorized recreation in US national parks signals the need for a better understanding of the meanings of these experiences. The fundamental issue facing many parks is whether snowmobiling is an appropriate activity. The matter of appropriateness is anchored in two questions: (1) Does snowmobiling in national parks cause unacceptable biophysical and social impacts? (2) Are snowmobiling experiences consistent with the fundamental purposes of national parks? This article particularly addresses the appropriateness issue and presents the findings of an interpretive study of the meanings of snowmobiling experiences in Yellowstone National Park (YNP). Sixty-five semistructured interviews were conducted with winter visitors who used snowmobiles in the park during the winter season of 1999. Overall, the study findings reveal that visitors on snowmobiles view YNP as a place to experience its wildland attributes, such as natural scenery, geothermal features, and, especially, wildlife. Snowmobiling is perceived as a mode of transportation rather than the experience in and of itself. The results challenge the popular image of snowmobiles as thrill-craft and snowmobilers as thrill-seekers. The study demonstrates that snowmobiling in YNP affords highly meaningful recreational experiences, grounded in appreciating the park's unique natural features and attributes. PMID- 15902455 TI - Nutrient-balance modeling as a tool for environmental management in aquaculture: the case of trout farming in France. AB - The control and prevention of nutrient pollution from fish farming plays an essential role in the French regulatory framework. Assessing nutrient emissions from fish farms is important in terms of farm authorization, taxation, and monitoring. Currently employed strategies involve both water sampling and empirical modeling. This article reports the work and outcomes of an expert panel that evaluated existing methodologies and their possible alternatives. The development and evaluation of a nutrient-balance approach was assessed as a potential alternative to currently used methodologies. A previously described nutrient-balance model was suggested and parameterized using expert choice, and its validity and applicability were assessed. The results stress that the nutrient-balance model provides more robust and relatively conservative waste estimates compared to the currently used methodologies. Sensitivity of the approach to the uneven data quality available at farm level, difficulties of on farm measurements, as well as model requirements and limitations are discussed. PMID- 15902456 TI - Information network topologies for enhanced local adaptive management. AB - We examined the principal effects of different information network topologies for local adaptive management of natural resources. We used computerized agents with adaptive decision algorithms with the following three fundamental constraints: (1) Complete understanding of the processes maintaining the natural resource can never be achieved, (2) agents can only learn by experimentation and information sharing, and (3) memory is limited. The agents were given the task to manage a system that had two states: one that provided high utility returns (desired) and one that provided low returns (undesired). In addition, the threshold between the states was close to the optimal return of the desired state. We found that networks of low to moderate link densities significantly increased the resilience of the utility returns. Networks of high link densities contributed to highly synchronized behavior among the agents, which caused occasional large-scale ecological crises between periods of stable and high utility returns. A constructed network involving a small set of experimenting agents was capable of combining high utility returns with high resilience, conforming to theories underlying the concept of adaptive comanagement. We conclude that (1) the ability to manage for resilience (i.e., to stay clear of the threshold leading to the undesired state as well as the ability to re-enter the desired state following a collapse) resides in the network structure and (2) in a coupled social-ecological system, the system-wide state transition occurs not because the ecological system flips into the undesired state, but because managers lose their capacity to reorganize back to the desired state. PMID- 15902457 TI - Long-term impacts of land-use change on non-point source pollutant loads for the St. Louis metropolitan area, USA. AB - A land-use-change simulation model (LEAM) and a non-point-source (NPS) water quality model (L-THIA) were closely coupled as LEAMwq in order to determine the long-term implications of various degree of urbanization on NPS total nitrogen (TN), total suspended particles (TSP), and total phosphorus (TP) loads. A future land-use projection in the St. Louis metropolitan area from 2005 to 2030 using three economic growth scenarios (base, low, and high) and a long-term precipitation dataset were used to predict the mean annual surface runoff and mean annual NPS pollutant loads in the region. Results show mean annual TN increases of 0.21%, 0.13%, and 0.14% by 2030 compared to 2000 under the base, high, and low scenarios, respectively. TSP and TP showed similar trends with different magnitudes. Corresponding changes in annual mean surface runoff were shown to be lower than expected, which might be attributed to the small-scale conversion pattern of land uses. In the most dramatic change (high growth) scenario, the runoff would increase across time but at varying rates, and temporal pollutant loads would result in a more complicated pattern than in the other scenarios. This is attributed to the complex interactions between event mean concentrations of pollutants and the magnitude of changes in land-use acre ages. By integrating L-THIA with LEAM, LEAMwq was found to be a useful planning tool to illustrate in a quick and simple manner how future water quality is connected to decision-making on future land-use change. PMID- 15902458 TI - An eco-balance of a recycling plant for spent lead-acid batteries. AB - This study applies Life Cycle Assessment (LCA) methodology to present an eco balance of a recycling plant that treats spent lead-acid batteries. The recycling plant uses pyrometallurgical treatment to obtain lead from spent batteries. The application of LCA methodology (ISO 14040 series) enabled us to assess the potential environmental impacts arising from the recycling plant's operations. Thus, net emissions of greenhouse gases as well as other major environmental consequences were examined and hot spots inside the recycling plant were identified. A sensitivity analysis was also performed on certain variables to evaluate their effect on the LCA study. The LCA of a recycling plant for spent lead-acid batteries presented shows that this methodology allows all of the major environmental consequences associated with lead recycling using the pyrometallurgical process to be examined. The study highlights areas in which environmental improvements are easily achievable by a business, providing a basis for suggestions to minimize the environmental impact of its production phases, improving process and company performance in environmental terms. PMID- 15902459 TI - Identification of land-cover changes through image processing and associated impacts on water reservoir conditions. AB - A temporal assessment of land-cover changes of the province Beykoz in Istanbul has been documented in this article. The study focuses on the acquisition and analysis of LANDSAT 5 TM images that reflect the drastic land-cover changes between the years 1984 and 2001 utilizing aerial photographs, orthophoto maps, standard topographic maps, and ground truth measurements. The status of the province, including its surrounding villages, were examined together with more specialized maps showing only the city center and part of the province that lies within the watershed of the Elmali Drinking Water Reservoir. The land distribution profiles were also calculated for each of the images to help authorities and decision-makers to better understand the main causes of such remarkable changes and to inform them of the changing quality conditions of the reservoir. Rapid, uncontrolled, and illegal urbanization accompanied by insufficient infrastructure has caused degradation of forests and barren lands in the province, especially within the past two decades. The proximity of the province to the reservoir's watershed, downtown Istanbul, and the transportation network has accelerated the land-cover changes whose adverse impacts on the reservoir water quality are sensed. It is intended that the data gathered and processed in this study will provide a basis for future sustainable urban planning and management activities. PMID- 15902460 TI - Typhlitis associated with Candida albicans and Pseudomonas aeruginosa infection in a patient with herbal drug-induced neutropenia. PMID- 15902461 TI - Biweekly oxaliplatin combined with oral capecitabine (OXXEL regimen) as first line treatment of metastatic colorectal cancer patients: a Southern Italy Cooperative Oncology Group phase II study. AB - Oxaliplatin 100 mg/m(2) iv on day 1, and capecitabine 1,000 mg/m(2) orally bid from day 1 (evening) to day 11 (morning) were administered every 2 weeks (OXXEL regimen) to 38 patients as first-line treatment for metastatic colorectal carcinoma. A total of 318 cycles were administered, with a median of 8 (range, 4 12) cycles per patient. Response rate (RR) was 45% (95% confidence interval (CI), 29%-62%), with 7 complete responses and 10 partial responses; furthermore, 12 patients showed a stable disease, so that a disease control was achieved in 29 (76%) patients. RR was greater among patients with performance status 0 (52%), without weight loss (52%), younger than 65 years (50%), and previously unexposed to adjuvant chemotherapy (48%), while no correlation was found with the actually delivered oxaliplatin dose intensity. Overall, haematological side effects were negligible, with no case of grade 4 toxicity, and only one patient suffering from an episode of grade 3 neutropenic fever. Severe anaemia occurred in 4 (11%) patients, and grade 3 neuropathy affected 9 (24%) patients. Median progression free survival was 7.9 (95% CI, 6.2-9.6) months, and median overall survival has not been reached yet. In conclusion, the OXXEL regimen resulted safe and active, and it deserves further evaluation in metastatic colorectal cancer patients. PMID- 15902462 TI - Antiangiogenic and antitumoral properties of a polysaccharide isolated from the seaweed Sargassum stenophyllum. AB - The potential antiangiogenic and antitumoral properties of SargA, a polysaccharide extracted from the brown marine alga Sargassum stenophyllum, were studied in assays carried out in chick embryos and mice. Gelfoam plugs containing SargA (2-1500 microg/plug) implanted in vivo into fertilized 6-day-old chicken eggs induced dose-related antiangiogenic activity in the chorioallantoic membrane (CAM). By day 8, the highest dose of SargA alone decreased the vessel number in the CAM by 64%, but coadministered with hydrocortisone (156 microg/plug, which alone caused 30% inhibition) failed to potentiate its antiangiogenic effect. Combined with basic fibroblast growth factor (50 ng/plug), SargA (1500 microg/plug) abolished angiogenesis stimulated by this factor in both chick embryo CAM and in subcutaneous (s.c.) Gelfoam plugs implanted in the dorsal skin of Swiss mice (measured as plug hemoglobin content). Repeated s.c. injections of SargA (1.5 or 150 microg per animal per day for 3 days) close to B16F10 melanoma cell tumors in the dorsal skin of mice markedly decreased tumor growth in a dose related fashion (by 40% and 80% at 2 weeks after the first injection, respectively), without evident signs of toxicity. SargA caused graded inhibitions of migration and viability of cultured B16F10 cells and also displayed antithrombotic activity in human plasma (5 mg/ml increased thrombin time 2.5-fold relative to saline). Thus, SargA exhibits pronounced antiangiogenic as well as antitumoral properties. Although the latter action of SargA might be related to the inhibition of angiogenesis, the polysaccharide also exerts cytotoxic effects on tumor cells. Because of its chemical characteristics and polyanionic constituents, we postulate that the polysaccharide SargA might modulate the activity of heparin-binding angiogenic growth factors. PMID- 15902463 TI - Production of catalases by Aspergillus niger isolates as a response to pollutant stress by heavy metals. AB - Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As(5+), Cd(2+), Cu(2+)) at final concentrations of 25 and 50 mg/L and H(2)O(2) (20 or 40 mM) mostly stimulated production of catalases only in isolates from mines surroundings, and H(2)O(2) and Hg(2+) caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H(2)O(2), as monitored by growth, than did the strain from the culture collection. PMID- 15902464 TI - Regulation of the ubiquinone (coenzyme Q) biosynthetic genes ubiCA in Escherichia coli. AB - Ubiquinone (Coenzyme Q) is an essential component of bacterial respiratory chains. The first committed step in the biosynthetic pathway is the formation of 4-hydroxybenzoate from chorismate by the enzyme chorismate pyruvate-lyase encoded by the ubiC gene. The 4-hydroxybenzoate is prenylated by 4-hydroxybenzoate octaprenyltransferase encoded by the ubiA gene. The two genes are linked at 91.5 min in the Escherichia coli chromosome. To study the regulation, operon fusions were constructed between these two genes and the lacZ gene. The fusions were introduced into the chromosome as a single copy at the lambda attachment site. Expression of beta-galactosidase was determined in strains carrying the operon fusions ubiC'-lacZ(+) ubiCA'-lacZ(+), and ubiA'-lacZ(+). In glycerol media, the highest level of expression was observed with the operon fusion ubiC'-lacZ(+). Compared with the ubiC'-lacZ(+), the ubiCA'-lacZ(+) operon fusion showed 26% of the activity while the ubiA'-lacZ(+) operon fusion had an activity of 1%. Thus, the ubiC gene is regulated by the upstream promoter while the ubiA gene lacks its own promoter. The effect of fermentable and oxidizable carbon sources on the expression of ubiC'-lacZ(+) was determined. The expression was low in the case of a fermentable carbon source, glucose, while in the presence of oxidizable carbon sources the expression increased 2- to 3-fold. When the expression of ubiC' lacZ(+) and ubiCA'-lacZ(+) operon fusions were compared under a wide variety of conditions, the levels of beta-galactosidase varied coordinately, suggesting that the ubiCA genes are organized into an operon. The variations in transcription of the operon under different nutritional conditions and in the regulatory mutants, arcA, fnr, and narXL are presented. PMID- 15902465 TI - Phylogenetic relationships of Xylella fastidiosa strains based on 16S-23S rDNA sequences. AB - In spite of the lack of resolution of Xylella fastidiosa phylogenetic relationships, parsimony analysis of the 16S-23S rDNA sequence from a wide range of hosts has been evaluated in this research. In order to establish an easier method for sequencing the spacer region completely, a new primer pair was designed. The sequences obtained revealed a higher level of variation than that found in 16S gene sequences, with similarity values ranging from 0.80 to 1.00. The cladogram constructed allowed the clustering of two major clades. From these results it has been possible to recognize the monophyletic grouping of some strains belonging to the same host, possibly representing only one infection process. However, for other hosts there is paraphyletic and polyphyletic grouping. This methodology followed from promising results regarding strain-host clustering. With the parsimony approach, hypothetical genealogical relationship among Xylella strains may be inferred. PMID- 15902466 TI - Cloning and characterization of a carboxylesterase from Bacillus coagulans 81-11. AB - A genomic library of Bacillus coagulans strain 81-11 was screened in Escherichia coli JM83 for lipolytic activity by using tributyrin agar plates. A 2.4 kb DNA fragment was subcloned from a lipolytic-positive clone and completely sequenced. Nucleotide sequence analysis predicted a 723 bp open reading frame (ORF), designated estC1, encoding a protein of 240 amino acids with an estimated molecular mass of 27,528 Da and a pI of 9.15. The deduced amino acid sequence of the estC1 gene exhibited significant amino acid sequence identity with carboxylesterases from thermophilic Geobacillus spp. and sequence analysis showed that the protein contains the signature G-X-S-X-G included in most esterases and lipases. Enzyme assays using p-nitrophenyl (p-NP) esters with different acyl chain lengths as the substrate confirmed the esterase activity. EstC1 exhibited a marked preference for esters of short-chain fatty acids, yielding the highest activity with p-NP butyrate. Maximum activity was found at pH 8 and 50 degrees C, although the enzyme displayed stability at temperatures up to 60 degrees C. PMID- 15902467 TI - Antibiotic susceptibility of Lactobacillus and Bifidobacterium species from the human gastrointestinal tract. AB - One hundred and twenty-two strains of Bifidobacterium and Lactobacillus species have been tested against 12 antibiotics and two antibiotic mixtures by a commercial system (Sensititre Anaero3; Treck Diagnostic Systems). The upper limits of some minimum inhibitory concentrations (MICs) were completed on MRS agar plates by the NCCLS procedure. All strains were sensitive to chloramphenicol and imipenem and most of the strains were resistant to metronidazole. Bifidobacteria isolates were susceptible to cefoxitin, whereas about half of the lactobacilli were resistant. Approximately 30% of the Bifidobacterium isolates were resistant to tetracycline, as well as five Lactobacillus strains belonging to four different species. None of the tested Bifidobacterium isolates was resistant to vancomycin, whereas a species-dependent resistance was found among the lactobacilli. Single strains of Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Lactobacillus acidophilus, Lactobacillus rhamnosus, and Lactobacillus brevis were resistant to erythromycin and/or clindamycin. Most of the observed resistances seemed to be intrinsic, but some others could be compatible with transmissible determinants. PMID- 15902468 TI - Stimulation of bacteriocin production by dialyzed culture media from different lactic acid bacteria. AB - The cross-effects of dialyzed postincubates (with a cut-off at 1000 Da) on the biomass and bacteriocin production of six strains of lactic acid bacteria were studied, and a predominance of stimulating responses was found, the characteristics of which suggested merely nutritional effects or the presence of precursor fragments of the bacteriocins. Additionally, cluster analysis of the detected responses provides an approach to define groups of highly compatible (potential consortia) or doubtfully compatible strains of lactic acid bacteria. Such a definition, which does not claim taxonomic value, has practical interest, however, in cases (e.g., silage production) in which it is convenient to use mixed inocula including strains able to establish positive interactions. PMID- 15902469 TI - Purification and characterization of a novel alpha-agarase from a Thalassomonas sp. AB - An agar-degrading Thalassomonas bacterium, strain JAMB-A33, was isolated from the sediment off Noma Point, Japan, at a depth of 230 m. A novel alpha-agarase from the isolate was purified to homogeneity from cultures containing agar as a carbon source. The molecular mass of the purified enzyme, designated as agaraseA33, was 85 kDa on both SDS-PAGE and gel-filtration chromatography, suggesting that it is a monomer. The optimal pH and temperature for activity were about 8.5 and 45 degrees C, respectively. The enzyme had a specific activity of 40.7 U/mg protein. The pattern of agarose hydrolysis showed that the enzyme is an endo-type alpha agarase, and the final main product was agarotetraose. The enzyme degraded not only agarose but also agarohexaose, neoagarohexaose, and porphyran. PMID- 15902470 TI - Replacement of active-site residues of quinoline 2-oxidoreductase involved in substrate recognition and specificity. AB - Amino acid residues in the active site of quinoline 2-oxidoreductase (Qor) that are deemed important for substrate binding and turnover were replaced by site directed mutagenesis. The apparent k(cat) values for quinoline were reduced 2.4-, 38-, 40-, and 199-fold in the protein variants QorA259G, QorW331G, QorV373A, and QorA546G, respectively. The substitution A259G did not significantly alter K(m app). Despite the presumed crucial role of W331 and V373 in substrate positioning, the replacements W331G (K(m app): 0.33 mM) and V373A (K(m app): 0.41 mM) only slightly affected affinity for quinoline (K(m app) of Qor: 0.12 mM). QorA546G showed an increased affinity for quinoline and quinoxaline, as suggested by its 4.3- and 7.5-fold decrease in K(m) (app (quinoline))and K(m app (quinoxaline)), respectively, compared with Qor. The relative activities of the protein variants towards substituted quinolines differed from those of Qor. QorW331G, for example, may be suitable for hydroxylation of quinoxaline and C4 substituted quinolines. PMID- 15902471 TI - Expression of pathogenicity-related genes of Xylella fastidiosa in vitro and in planta. AB - Xylella fastidiosa is responsible for several economically important plant diseases. It is currently assumed that the symptoms are caused by vascular occlusion due to biofilm formation. Microarray technology was previously used to examine the global gene expression profile of X. fastidiosa freshly isolated from symptomatic plants or after several passages by axenic culture medium, and different pathogenicity profiles have been obtained. In the present study the expression of some pathogenicity-related genes was evaluated in vitro and in planta by RT-PCR. The results suggest that adhesion is important at the beginning of biofilm formation, while the genes related to adaptation are essential for the organism's maintenance in planta. Similar results were observed in vitro mainly for the adhesion genes. The pattern of expression observed suggests that adhesion modulates biofilm formation whereas the expression of some adaptation genes may be related to the environment in which the organism is living. PMID- 15902472 TI - Piboserod (SB 207266), a selective 5-HT4 receptor antagonist, reduces serotonin potentiation of neurally-mediated contractile responses of human detrusor muscle. AB - The aim of this study is to evaluate the potency of piboserod (SB 207266), a selective 5-HT(4) receptor antagonist, at inhibiting the 5-HT(4)-mediated potentiating effect of serotonin (5-HT) on the neurally-mediated contractile responses of human detrusor strips to electrical field stimulations (EFS). Strips of human detrusor muscle were mounted in Krebs-HEPES buffer under a resting tension of 500 mg and EFS (20 Hz, 1 ms duration at 300 mA for 5 s) was applied continuously at 1 min intervals. After stabilization of the EFS-induced contractions, concentration-response curves to 5-HT (0.1 nM-100 microM) were constructed in the absence or presence of 1 or 100 nM of piboserod. The experiments were performed in the presence of methysergide (1 microM) and ondansetron (3 microM) to block 5HT(1)/5HT(2) and 5-HT(3) receptors, respectively. 5-HT potentiated the contractile responses to EFS of human bladder strips in a concentration-dependent manner, with a maximum mean of 60.0+/-19.9% of the basal EFS-evoked contractions. Piboserod did not modify the basal contractions but concentration-dependently antagonized the ability of 5-HT to enhance bladder strip contractions to EFS. In presence of 1 and 100 nM of piboserod, the maximal 5-HT-induced potentiations were reduced to 45.0+/-7.9 and 38.7+/-8.7%, respectively. A mean apparent antagonist dissociation constant value (K(B)) of 0.56+/-0.09 nM was determined. These data show the ability of piboserod to antagonize with high potency the enhancing properties of 5-HT on neurally mediated contractions of isolated human bladder strips. Therefore, the 5-HT(4) receptor might represent an attractive pharmacological target for the treatment of overactive bladder. PMID- 15902473 TI - Ejaculatory disorders: epidemiology and current approaches to definition, classification and subtyping. AB - Ejaculatory disorders (disorders of emission, ejaculation and orgasm) are the most frequent sexual complaint. Conventional algorithms on ejaculatory disorders are based on an organic or psychogenic dichotomy, with the latter being traditionally considered the main etiological cause. The scope of this review is to propose a new classification of ejaculatory disorders, with special emphasis on epidemiology and subtyping of the most frequent: premature ejaculation (PE). There is growing agreement on definition, diagnosis, and treatment options. In many cases, ejaculatory disorders can be classified by psycho-neuro-uro-endocrine symptoms. Epidemiological data, new classifications and subtyping, together with new diagnostic procedures and the availability of pharmacological aids, place this topic in the mainframe of sexual medicine. This will soon demolish the barriers to seeking help perceived by patients suffering ejaculatory disorders. PMID- 15902474 TI - Cochlear function in mice following inhalation of brevetoxin-3. AB - Brevetoxin-3 was shown previously to adversely affect central auditory function in goldfish. The present study evaluated the effects of exposure to this agent on cochlear function in mice using the 2f(1)-f(2) distortion-product otoacoustic emission (DPOAE). Towards this end, inbred CBA/CaJ mice were exposed to a relatively high concentration of brevetoxin-3 (approximately=400 microg/m(3)) by nose-only inhalation for a 2-h period. Further, a subset of these mice received a second exposure a day later that lasted for an additional 4 h. Mice exposed only once for 2 h did not exhibit any notable cochlear effects. Similarly, mice exposed two times, for a cumulative dose of 6 h, exhibited essentially no change in DPOAE levels. PMID- 15902475 TI - Peritoneal taurolidine lavage in children with localised peritonitis due to appendicitis. AB - Despite aggressive surgical treatment, rational antibiotic therapy, and modern intensive care, generalised peritonitis remains a major threat in the paediatric age group. Several adjuvant strategies such as peritoneal saline lavage and peritoneal drainage have been utilised. Taurolidine, derived from the amino acid taurine, has bactericidic, antiendotoxic, and antiinflammatory properties. It has been introduced previously for intraoperative peritoneal lavage in treating peritonitis in adults. The aim of our study was to evaluate the effect of peritoneal taurolidine lavage on the clinical course and serological inflammation markers in children with perforated appendicitis and localised peritonitis. A series of 27 children presenting with appendicitis between January 1999 and July 2001 were included in the study after parental informed consent. All patients underwent open appendectomy. Taurolidine peritoneal lavage was applied in 15 randomly selected children (eight girls and seven boys; mean age 10 years and 10 months). Twelve children received saline peritoneal lavage and served as the control group (six girls and six boys; mean age 9 years and 7 months). Blood was taken preoperatively and on postoperative days 1, 3, 7, and 14. Full blood cell count, C-reactive protein, endotoxin, interleukin-1, interleukin-6, soluble interleukin-2 receptor, tumour necrosis factor alpha, and procalcitonin were investigated to evaluate the serological course of inflammation. Both groups initially presented with severe inflammation as evidenced clinically and serologically. The clinical postoperative course was uneventful in 13/15 patients in the treatment group and 10/12 patients in the control group. The remaining patients presented complications: intraperitoneal abscess or early postoperative bowel obstruction. With regard to the serological inflammatory parameters, no significant differences were found between the two groups except for the soluble interleukin-2-receptor on the 7th postoperative day. In conclusion, the expected reduction of endotoxin levels and inflammatory activity in the treatment group was not evident. A significant advantage of adjuvant peritoneal taurolidine lavage in the surgical therapy of children with localised peritonitis due to appendicitis could not be shown in our study. PMID- 15902476 TI - Perineal hamartoma with accessory scrotum, anorectal anomaly, and hypospadias-a rare association with embryological significance: case report. AB - A rare case of a newborn male with a perineal hamartoma, accessory scrotum, anorectal anomaly, hypospadias, and bifid scrotum is reported, with discussion of its embryological significance. Only three other cases with such a combination of anomalies have been reported in the English literature. PMID- 15902477 TI - Herniation through the foramen of Winslow presenting as obstructive jaundice. AB - Herniation through the foramen of Winslow is a rare variety of paraduodenal hernia. We report a 19-month-old child with obstructive jaundice due to midgut herniation through the foramen of Winslow with associated volvulus. The herniation and volvulus were precipitated by intestinal malrotation. The patient underwent extraamniotic silo repair of exomphalos major in the neonatal period. Investigation for malrotation is recommended after extraamniotic closure of exomphalos. PMID- 15902478 TI - Unexpected sudden death related to medullary brain lesions. PMID- 15902479 TI - Comparison between two methods of drain clamping after total knee arthroplasty. AB - INTRODUCTION: To study the clinical outcome of two methods of drain clamping after total knee arthroplasty and to determine the effect on blood loss and blood transfusion. PATIENTS AND METHODS: A prospective study involving 73 patients, randomized into two groups. Group 1 included patients in whom the drain was clamped for 1 h postoperatively, after which it was released and kept open for 48 h. Group 2 included patients in whom the drain was clamped and released for 10 min every 2 h for 24 h. The drain was removed at 48 h in both groups. Patient's demographic details, intraoperative and postoperative blood loss, and haemoglobin values on the preoperative and postoperative days (1, 4, 7, 14) were recorded. RESULTS: The mean postoperative drainage in group 1 was 520.65+/-170 ml, which was significantly higher than that of group 2, 367.62+/-141.1 ml (p < 0.05, Student's t-test). The drain charting shows 65% of drainage volume occurs in the first 8 h in both groups. The study suggests a reduction in the incidence of blood transfusion in the 2-hourly groups, although it was not statistically significant. There was a difference in the haemoglobin drop between both groups, but statistically the p value was marginally above 0.05 (p = 0.086) and hence not significant. CONCLUSION: The method of 2-hourly clamping of drain and release for 10 min significantly reduces postoperative blood loss, without any added increase of complication, after total knee arthroplasty. PMID- 15902480 TI - Histamine helps development of eczematous lesions in experimental contact dermatitis in mice. AB - Histamine is released from mast cells in the skin, causing urticaria and itching. However, little is known about the roles of histamine in development of eczematous lesions in contact dermatitis. Effects of histamine on development of eczematous lesions in contact dermatitis were assessed using histamine-deficient mice in which contact dermatitis was developed by repeated application of diphenylcyclopropenone. Development of eczematous lesions in contact dermatitis was suppressed in histamine-deficient mice compared to wild-type mice. H(1) agonist ((6-12-(4-imidazol)ethylamino)-N-(4-trifluoro- methylphenyl)hepatanecarboxamide) promoted development of eczematous lesions in histamine-deficient mice. H(1) receptor antagonist (loratadine) suppressed development of eczematous lesions in wild-type mice, whereas H(2) agonist (dimaprit) and receptor antagonist (cimetidine) were ineffective. These results suggest that histamine facilitates the development of eczematous lesions in a murine model of contact dermatitis via H(1) receptors. PMID- 15902481 TI - A rapid and definitive diagnosis of kerosene dermatitis by an analysis of detached lesional epidermis using gas chromatography-mass spectrometry. AB - A 73-year-old woman, who suffered from erythema with bullae and pustules on her abdomen and anterior right thigh, visited our hospital without an awareness of the causative agents. The lesions appeared like first and second degree burns. The small amount of detached roof of bulla was sampled without skin biopsy before the ointment treatment. The sample was sonicated in an ultrasonic bath for 1 min in n-pentane, and then 1 mul of the extract was analyzed by gas chromatography mass spectrometry (GC-MS). The causative agent was determined to be kerosene. An examination of blood samples collected at the first visit failed to detect kerosene, though traces of trimethylbenzene were detected. A GC-MS analysis of the small sample of lesional epidermis was very useful to identify kerosene as a causative agent. PMID- 15902482 TI - Control of MAP kinase signaling to the nucleus. AB - MAP kinase (MAPK) signaling is among central signaling pathways that regulate cell proliferation, cell differentiation and apoptosis. As MAPK should transmit extracellular signals to proper regions or compartments in cells, controlling subcellular localization of MAPK is important for regulating fidelity and specificity of MAPK signaling. The ERK1/2-type of MAPK is the best characterized member of the MAPK family. In response to extracellular stimulus, ERK1/2 translocates from the cytoplasm to the nucleus by passing through the nuclear pore by several independent mechanisms. Sef (similar expression to fgf genes), a transmembrane protein, has been shown to be a regulator of subcellular distribution of ERK1/2. Sef binds to activated MEK1/2, the specific activator of ERK1/2, and tethers the activated MEK1/2/activated ERK1/2 complex to the Golgi apparatus and the plasma membrane. Thus, Sef blocks ERK1/2 signaling to the nucleus and allows signaling to the cytoplasm. Here we review recent findings on spatial regulation of MAPK, especially on nucleocytoplasmic trafficking of ERK1/2. PMID- 15902483 TI - Polybrominated diphenyl ethers--plasma levels and thyroid status of workers at an electronic recycling facility. AB - OBJECTIVES: Personnel working with electronic dismantling are exposed to polybrominated diphenyl ethers (PBDEs), which in animal studies have been shown to alter thyroid homeostasis. The aim of this longitudinal study was to measure plasma level of PBDEs in workers at an electronic recycling facility and to relate these to the workers' thyroid status. METHODS: PBDEs and three thyroid hormones: triiodothyronine (T(3)), thyroxin (T(4)) and thyroid stimulating hormone (TSH) were repeatedly analysed in plasma from 11 workers during a period of 1.5 years. RESULTS: Plasma levels of PBDEs at start of employment were <0.5 9.1 pmol/g lipid weight (l.w.). The most common congener was PBDE #47 (median 2.8 pmol/g l.w.), followed by PBDE #153 (median 1.7 pmol/g l.w.), and PBDE #183 had a median value of <0.19 pmol/g l.w. After dismantling the corresponding median concentrations were: 3.7, 1.7 and 1.2 pmol/g l.w., respectively. These differences in PBDE levels were not statistically significant. PBDE #28 showed a statistically significantly higher concentration after dismantling than at start of employment (P=0.016), although at low concentrations (start 0.11 pmol/g l.w. and dismantling 0.26 pmol/g l.w.). All measured levels of thyroid hormones (T(3), T(4) and TSH) were within the normal physiological range. Statistically significant positive correlations were found between T(3) and #183 in a worker, between T(4) and both #28 and #100 in another worker and also between TSH and #99 and #154 in two workers. CONCLUSIONS: The workers' plasma levels of PBDEs fluctuated during the study period. Due to small changes in thyroid hormone levels it was concluded that no relevant changes were present in relation to PBDE exposure within the workers participating in this study. PMID- 15902484 TI - Tumor induced local fibrogenic effect by hepatic metastasis of insulinoma. PMID- 15902485 TI - Cytoplasmic expression of c-erbB2 in non-small cell lung cancers. AB - The aim of this study was to investigate the expression of c-erbB2 in non-small cell lung cancers (NSCLCs) with attention both to membranous and cytoplasmic reaction, and to try to elucidate the meaning of cytoplasmic expression of c erbB2 in NSCLCs. Immunohistochemical c-erbB2 expression and related clinico pathological features were examined in 312 surgically resected patient tissues of NSCLCs, including 175 cases of adenocarcinoma and 137 cases of squamous cell carcinoma. Immunostaining of inner- and ecto-domain of c-erbB2, mRNA expression and the quantitation of soluble c-erbB2 in cultured media were performed in five NSCLC cell lines. Cytoplasmic expression of c-erbB2 was observed more frequently than membranous, both in patient tissues and cell lines. Neither membranous nor cytoplasmic expression of c-erbB2 was significantly correlated with short outcome in NSCLCs. Membranous c-erbB2 was expressed by both inner and ecto-domain, while cytoplasmic c-erbB2 was expressed by either or both inner and ecto-domain. c erbB2 mRNA was produced in most cell lines; however, the soluble form was only detectable in a cell line that only presented a membranous c-erbB2. In conclusion, cytoplasmic c-erbB2 of NSCLCs was not a full-length protein only expressed in cellular membrane, but reflected degenerated c-erbB2 fragments with less functional ability. PMID- 15902486 TI - Type and prevalence of BRAF mutations are closely associated with papillary thyroid carcinoma histotype and patients' age but not with tumour aggressiveness. AB - A high prevalence of the BRAF(V600E) somatic mutation was recently reported in several series of papillary thyroid carcinomas (PTC). This mutation appears to be particularly prevalent in PTC with a predominantly papillary architecture. Another BRAF mutation (K601E) was detected in a follicular adenoma and in some cases of the follicular variant of PTC. The few studies on record provided controversial data on the relationship between the occurrence of BRAF mutations and clinicopathologic parameters such as gender, age and tumour staging. In an attempt to clarify such controversies we decided to enlarge our previous series to 315 tumours or tumour-like lesions diagnosed in 280 patients, including a thorough analysis of several clinicopathologic features. The BRAF(V600E) mutation was exclusively detected in PTC with a papillary or mixed follicular/papillary architecture both of the conventional type (46%) and of other histotypes, such as microcarcinoma (43%), Warthin-like PTC (75%) and oncocytic variant of PTC (55%). The BRAF(K601E) mutation was detected in four of the 54 cases of the follicular variant of PTC (7%). The mean age of patients with conventional PTC harbouring BRAF(V600E) (46.7 years) was significantly higher (P<0.0001) than that of patients with conventional PTC without BRAF(V600E) (29.5 years). The BRAF (BRAF(V600E)) mutated PTC did not exhibit signs of higher aggressiveness (size, vascular invasion, extra-thyroid extension and nodal metastasis) and were in fact less often multicentric than PTC without the mutation. PMID- 15902487 TI - Cellular localisation of survivin: impact on the prognosis in colorectal cancer. AB - PURPOSE: The present study was designed to determine whether the nuclear or cytoplasmic expression of survivin, was related to clinicopathological parameters and survival in sporadic colon carcinomas. METHODS: Western blotting of cell fractions and immunocytochemical methodology were used in five human colon cancer cell lines. Immunohistochemical study was performed in formalin-fixed paraffin embedded section from 46 patients with sporadic colorectal adenocarcinomas with a polyclonal antibody directed against survivin. Apoptotic index was evaluated by using the M30 antibody. Survival rates were estimated by the Kaplan-Meier method and compared using the log-rank test. Multivariate survival analysis was performed by the Cox proportional hazards model. RESULTS: Western blotting and immunocytochemistry analyses confirmed that survivin could be detected both in the nucleus and the cytoplasm. Immunohistochemical analysis demonstrated that 39% of tumours expressed survivin in the nucleus and 41% in the cytoplasm. No relationship was observed between survivin expression and clinicopathological features. Unexpectedly, the apoptotic index appeared to be linked with high survivin nuclear expression. Overall, 3-year observed survival rate was 73% in patients with cytoplasmic survivin expression versus 48% for negative expression (P = 0.14). Survival was 72% versus 50% for positive nuclear survivin expression versus negative (P = 0.16). After adjustment for age and stage, cytoplasmic survivin expression was a significant prognostic factor. A high level of expression was associated to a better survival: RR = 0.35 [0.13-0.98], P = 0.045. CONCLUSION: These results indicate that the analysis of the subcellular expression of survivin is a determining factor to define the prognostic value. Its evaluation, using a polyclonal antibody, might help clinicians in the stratification of patients with colorectal cancer. PMID- 15902488 TI - Cloning and characterization of a gene for an LRR receptor-like protein kinase associated with cotton fiber development. AB - Cotton fiber is an ideal model for studying plant cell elongation and cell wall biogenesis, but the genes that are critical for the regulation of fiber development are largely unknown. We report here the cloning and characterization of a receptor-like kinase gene (designated GhRLK1), expression of which is induced during the period of active secondary wall synthesis in the cotton fiber cells. We demonstrate that GhRLK1 is located in the plasma membrane and shows dual specificity as both a serine/threonine kinase and a tyrosine kinase. Our results suggest a possible role of GhRLK1 in the signal transduction pathway that is involved in the induction and maintenance of active secondary wall formation during fiber development. PMID- 15902489 TI - DNA microarray analysis of Methanosarcina mazei Go1 reveals adaptation to different methanogenic substrates. AB - Methansarcina mazei Go1 DNA arrays were constructed and used to evaluate the genomic expression patterns of cells grown on either of two alternative methanogenic substrates, acetate or methanol, as sole carbon and energy source. Analysis of differential transcription across the genome revealed two functionally grouped sets of genes that parallel the central biochemical pathways in, and reflect many known features of, acetate and methanol metabolism. These include the acetate-induced genes encoding acetate activating enzymes, acetyl-CoA synthase/CO dehydrogenase, and carbonic anhydrase. Interestingly, additional genes expressed at significantly higher levels during growth on acetate included two energy-conserving complexes (the Ech hydrogenase, and the A1A0-type ATP synthase). Many previously unknown features included the induction by acetate of genes coding for ferredoxins and flavoproteins, an aldehyde:ferredoxin oxidoreductase, enzymes for the synthesis of aromatic amino acids, and components of iron, cobalt and oligopeptide uptake systems. In contrast, methanol-grown cells exhibited elevated expression of genes assigned to the methylotrophic pathway of methanogenesis. Expression of genes for components of the translation apparatus was also elevated in cells grown in the methanol medium relative to acetate, and was correlated with the faster growth rate observed on the former substrate. These experiments provide the first comprehensive insight into substrate-dependent gene expression in a methanogenic archaeon. This genome-wide approach, coupled with the complementary molecular and biochemical tools, should greatly accelerate the exploration of Methanosarcina cell physiology, given the present modest level of our knowledge of these large archaeal genomes. PMID- 15902490 TI - Analysis of the melon genome in regions encompassing TIR-NBS-LRR resistance genes. AB - Plant genomes contain numerous genes (R-genes) that play a role in initiating defence measures against their particular pathogens. Defence mechanisms controlled by R-genes have been the focus of extensive research over the past several years. The majority of the R-genes described so far belong to a super family of genes (150-600 members) that encode proteins with a nucleotide binding site (NBS), some leucine-rich repeats (LRR) and an N-terminal domain that shows similarity to the Toll and Interleukin-1 receptors (TIR) or a N-terminal coiled coil (CC) domain. Analysis of four regions of the melon (Cucumis melo) genome, including two sequenced BACs, identified 14 TIR-NBS-LRR genes. Known disease resistance genes have been mapped in three of these regions. Transcriptional expression was detected for predicted genes that are possibly involved in defence responses to pathogen attack. TIR-NBS-LRR genes appear to be clustered in the melon genome. They contain all the conserved motifs that have previously been described for their counterparts in other species, although differences were also detected. The results presented here may contribute to a better understanding of the genomic distribution and evolution of this group of resistance gene homologues and their variability. PMID- 15902491 TI - Characterization of in vivo recombination activities in the mouse embryo. AB - Homologous recombination makes use of sequence homology to repair DNA and to rearrange genetic material. In mammals, these processes have mainly been characterized using cultured cell systems. We have developed an assay that allows us to quantitatively analyze homologous recombination in vivo in the mouse embryo. Transgenic mouse lines were generated by microinjection into a fertilized mouse ovum of a vector containing two homologous LINE-1 (L1) sequences arranged as a direct repeat: these sequences can recombine with each other and with endogenous L1 sequences before, during or after integration of the vector into the genome. Using a plasmid rescue procedure, we determined the composition of the integrated vector array in several transgenic mice and their descendants. Homologous recombination frequencies were found to be strikingly high, involving 70% of integrated vectors in some arrays, with homologous deletions being five times more frequent than gene conversion without crossing-over. Interestingly, non-homologous recombination was found to be much less frequent. We also found that endogenous L1 sequences could be involved in homologous recombination events in the mouse embryo, and that the integrated arrays could be modified from generation to generation by homologous recombination between the integrated L1 sequences. PMID- 15902492 TI - The Elp3 subunit of human Elongator complex is functionally similar to its counterpart in yeast. AB - Functions of the Elp3 subunit of the recently purified human Elongator were studied using an in vivo yeast complementation system. We demonstrated that the human ELP3 gene (hELP3) was able partially to complement functional defects of yeast elp3Delta cells. Furthermore, a chimeric ELP3 gene (yhELP3) encoding a protein in which the putative histone acetyltransferase (HAT) domain of hELP3 fused to the remainder of the yeast Elp3p corrected the growth defects of elp3Delta cells and complemented the slow activation of some inducible genes. Moreover, deletion of the B motif of the catalytic domain of the HAT region of hELP3 eliminated the ability of yhELP3 to complement elp3Delta in vivo, indicating that the HAT activity is essential for ELP3 function. We also demonstrated that replacement of specific lysine residues in histones H3 and H4 by arginine affected the complementation capacity of both the yeast gene (yELP3) and the chimeric yhELP3 in the elp3Deltastrain. Specifically, mutation of lysine 14 of H3 (H3 K14R) or lysine-8 of H4 (H4 K8R) reduced the ability of yELP3 and yhELP3 to complement the elp3Delta mutant, whereas simultaneous mutation of both sites (H3 K14R/H4 K8R) almost completely abolished complementation. These results imply a link between the acetylation of specific sites in nucleosomal histones and the regulation of transcription elongation by human Elp3. The data presented in this report suggest that the Elp3 subunits of human and yeast are highly conserved in their structure and functions. PMID- 15902493 TI - A genetic linkage map of the soybean cyst nematode Heterodera glycines. AB - A genetic linkage map of the soybean cyst nematode (SCN) Heterodera glycines was constructed using a population of F2 individuals obtained from matings between two highly inbred SCN lines, TN16 and TN20. The AFLP fingerprinting technique was used to genotype 63 F2 progeny with two restriction enzyme combinations (EcoRI/MseI and PstI/TaqI) and 38 primer combinations. The same F2 population was also genotyped for Hg-cm-1 (H. glycines chorismate mutase-1), a putative virulence gene, using real-time quantitative PCR. Some of the markers were found to be distributed non-randomly. Even so, of the 230 markers analyzed, 131 could be mapped onto ten linkage groups at a minimum LOD of 3.0, for a total map distance of 539 cM. The Hg-cm-1 locus mapped to linkage group III together with 16 other markers. The size of the H. glycines genome was estimated to be in the range of 630-743 cM, indicating that the current map represents 73-86% of the genome, with a marker density of one per 4.5 cM, and a physical/genetic distance ratio of between 124 kb/cM and 147 kb/cM. This genetic map will be of great assistance in mapping H. glycines markers to genes of interest, such as nematode virulence genes and genes that control aspects of nematode parasitism. PMID- 15902494 TI - Transforming growth factor-beta1 downregulates beating frequency and remodeling of cultured rat adult cardiomyocytes. AB - We have observed increased levels of transforming growth factor-beta1 (TGF-beta1) in human hibernating myocardium (HM). Impaired ventricular function in HM is known to be restored to normal following revascularization implying that myocardial structure in HM is to a certain degree preserved. We have therefore tested whether TGF-beta1 can imitate features of HM by reducing the number and frequency of beating cells (chronotropism) and structural remodeling of cultured adult rat cardiomyocytes (ARC), thus saving substrate, energy, and oxygen. Parameters measured were cell size, protein synthesis, protein degradation, protein content, myofibrillogenesis, and chronotropism. ARC were stimulated for 6 days with sera from patients with coronary heart disease, as this period led to a maximum response of cells. An increase of 90% in cell surface area following such treatment was reduced to a 20% increase of the original size by TGF-beta1. Concomitantly, the rate of protein synthesis dropped from 3.6-fold to 2.4-fold, and myofibrillogenesis was reduced. TGF-beta1 downregulated both the number of contracting cells from 81% to 10% and the frequency from 52 to nine beats per minute. However, TGF-beta1 treatment did not reduce the augmentation of protein content (1.28-fold versus 1.25-fold) indicating that protein degradation was also inhibited. Similar results were obtained with serum from healthy volunteers. The effects of TGF-beta1 were reversible. We conclude that TGF-beta1 constrains protein turnover and beating activity in underperfused myocardium, thus mediating protection by adapting myocytes to shortages in blood supply. PMID- 15902495 TI - Comparison of the localization of Bacillus thuringiensis Cry1A delta-endotoxins and their binding proteins in larval midgut of tobacco hornworm, Manduca sexta. AB - Tobacco hornworm, Manduca sexta, is a model insect for studying the action of Bacillus thuringiensis (Bt) Cry toxins on lepidopterans. The proteins, which bind Bt toxins to midgut epithelial cells, are key factors involved in the insecticidal functions of the toxins. Three Cry1A-binding proteins, viz., aminopeptidase N (APN), the cadherin-like Bt-R1, and membrane-type alkaline phosphatase (m-ALP), were localized, by immunohistochemistry, in sections from the anterior, middle, and posterior regions of the midgut from second instar M. sexta larvae. Both APN and m-ALP were distributed predominantly along microvilli in the posterior region and to a lesser extent on the apical tip of microvilli in the anterior and middle regions. Bt-R1 was localized at the base of microvilli in the anterior region, over the entire microvilli in the middle region, and at both the apex and base of microvilli in the posterior region. The localization of rhodamine-labeled Cry1Aa, Cry1Ab, and Cry1Ac binding was determined on sections from the same midgut regions. Cry1Aa and Cry1Ab bound to the apical tip of microvilli almost equally in all midgut regions. Binding of Cry1Ac was much stronger in the posterior region than in the anterior and middle regions. Thus, binding sites for Bt proteins and Cry1A toxins are co-localized on the microvilli of M. sexta midgut epithelial cells. PMID- 15902496 TI - Immunocytochemical localization of protein p27BBP in human skin and invertebrate (Sepia officinalis) integument. AB - The protein p27BBP (alias eIF6) occurs in yeast and mammalian epithelial cells. It is essential for ribosome genesis and has also been implicated in the functionality of integrins and intermediate filaments. By immunoblot, we show that homogenized integument from Sepia officinalis (Cephalopoda, Mollusca) contains a protein with immunological properties that closely resemble those of p27BBP. We also demonstrate, by immunogold electron microscopy with an indirect immunoreaction technique on ultrathin sections of human skin and Sepia integument, that p27BBP is constantly present in both species in epithelial cells, fibroblasts, and muscle fibers. It is found in the vicinity of intermediate filaments, in nucleoli, along the internal wall of the nuclear membrane, and in association with desmosomes and hemidesmosomes and occasionally occurs extracellularly. Thus, the structure and function of p27BBP seem to have been highly conserved throughout evolution; the protein appears to be essential in eukaryotic cells in which it interacts with several ultrastructural components of diverse function. PMID- 15902497 TI - Synaptic connections of cone bipolar cells that express the neurokinin 1 receptor in the rabbit retina. AB - We have investigated and further characterized, in the rabbit retina, the synaptic connectivity of the ON-type cone bipolar cells that are immunoreactive for an antibody against the neurokinin-1 receptor (NK1R). NK1R-immunoreactive bipolar cell axons terminate in stratum 4 of the inner plexiform layer. The axons of NK1R-positive bipolar cells receive synaptic inputs from amacrine cells through conventional synapses and from putative AII amacrine cells via gap junctions. The major outputs from NK1R-positive bipolar cells make contacts with amacrine cell processes. The most frequent postsynaptic dyads comprise two amacrine cell processes. Double-labeling experiments with antibodies against NK1R and either calretinin or glycine have demonstrated that NK1R-immunoreactive bipolar cells form gap junctions with AII amacrine cells. Thus, NK1R-positive cone bipolar cells, together with calbindin-positive cone bipolar cells, may play an important role in transferring rod signals to the ON-type ganglion cells of the cone pathway in the rabbit retina. PMID- 15902498 TI - P2X receptors in the rat uterine cervix, lumbosacral dorsal root ganglia, and spinal cord during pregnancy. AB - ATP, an intracellular energy source, is released from cells during tissue stress, damage, or inflammation. The P2X subtype of the ATP receptor is expressed in rat dorsal root ganglion (DRG) cells, spinal cord dorsal horn, and axons in peripheral tissues. ATP binding to P2X receptors on nociceptors generates signals that can be interpreted as pain from damaged tissue. We have hypothesized that tissue stress or damage in the uterine cervix during late pregnancy and parturition can lead to ATP release and sensory signaling via P2X receptors. Consequently, we have examined sensory pathways from the cervix in nonpregnant and pregnant rats for the presence of purinoceptors. Antiserum against the P2X3 receptor subtype showed P2X3- receptor immunoreactivity in axon-like structures of the cervix, in small and medium-sized neurons in the L6/S1 DRG, and in lamina II of the L6/S1 spinal cord segments. Retrograde tracing confirmed the projections of axons of P2X3-receptor-immunoreactive DRG neurons to the cervix. Some P2X3-receptor-positive DRG neurons also expressed estrogen receptor-alpha immunoreactivity and expressed the phosphorylated form of cyclic AMP response element-binding protein at parturition. Western blots showed a trend toward increases of P2X3-receptor protein between pregnancy (day 10) and parturition (day 22-23) in the cervix, but no significant changes in the DRG or spinal cord. Since serum estrogen rises over pregnancy, estrogen may influence purinoceptors in these DRG neurons. We suggest that receptors responsive to ATP are expressed in uterine cervical afferent nerves that transmit sensory information to the spinal cord at parturition. PMID- 15902499 TI - Expression of toll-like receptor 2 and 4 in lipopolysaccharide-induced lung injury in mouse. AB - Pattern recognition receptors, which include the toll-like receptors (TLRs), are considered to play an important role in the response against lipopolysaccharide (LPS). In this study, we performed a reverse transcriptase/polymerase chain reaction (RT-PCR) study, Western analysis, immunohistochemical staining, and RT PCR-amplified in situ hybridization of TLR2 and TLR4 in the case of LPS-induced lung injury. The expression of TLR2 and TLR4 increased in the lung rapidly after LPS inhalation and peaked at 24 h, followed by a gradual decrease. TLR2 and TLR4 expression was observed on the bronchial epithelium and tissue macrophages. In the early hours after inhalation of fluorescein-isothiocyanate (FITC)-labeled LPS, LPS was detected mainly on the bronchial epithelium and on a few of tissue macrophages. One day after inhalation, the LPS signals disappeared in the lungs of the mice, except for a few alveolar macrophages. The expression of TLR2, TLR4, and CD14 was coincident with the signals of FITC-labeled LPS. Instillation of liposome-encapsulated dichloromethylene diphosphonate induced a significant decrease in alveolar macrophages. In the macrophage-depleted mice, however, expression of TLR2 and TLR4 mRNA or protein was slightly suppressed in the lung after LPS inhalation. These data suggest that the bronchial epithelium and macrophages play crucial roles in LPS-induced lung injury through TLR2 and TLR4. PMID- 15902500 TI - Selective visualisation of neuroepithelial bodies in vibratome slices of living lung by 4-Di-2-ASP in various animal species. AB - Pulmonary neuroepithelial bodies (NEBs) are extensively innervated organoid groups of neuroendocrine cells that lie in the epithelium of intrapulmonary airways. Our present understanding of the morphology of NEBs is comprehensive, but direct physiological studies have so far been challenging because the extremely diffuse distribution of NEBs makes them inaccessible in vivo and because a reliable in vitro model is lacking. Our aim has been to optimise an in vitro method based on vibratome slices of living lungs, a model that includes NEBs, the surrounding tissues and at least part of their complex innervation. This in vitro model offers satisfactory access to pulmonary NEBs, provided that they can be differentiated from other tissue elements. The model was first optimised for living rat lung slices. Neutral red staining, reported to stain rabbit NEBs, proved unsuccessful in rat slices. On the other hand, the styryl pyridinium dye, 4-(4-diethylaminostyryl)-N-methylpyridinium iodide (4-Di-2-ASP), showed brightly fluorescent cell groups, reminiscent of NEBs, in the airway epithelium of living lung slices from rat. In addition, nerve fibres innervating the NEBs were labelled. The reliable and specific labelling of pulmonary NEBs by 4-Di-2-ASP was corroborated by immunostaining for protein gene-product 9.5. Live cell imaging and propidium iodide staining further established the acceptable viability of 4-Di-2-ASP-labelled NEB cells in lung slices, even over long periods. Importantly, the in vitro model and 4-Di-2-ASP staining procedure for pulmonary NEBs appeared to be equally reproducible in mouse, hamster and rabbit lungs. Diverse immunocytochemical procedures could be applied to the lung slices providing an opportunity to combine physiological and functional morphological studies. Such an integrated approach offers additional possibilities for elucidating the function(s) of pulmonary NEBs in health and disease. PMID- 15902501 TI - Selective inhibition of ventral temporal but not dorsal nasal neurites from mouse retinal explants during contact with chondroitin sulphate. AB - We have determined whether chondroitin sulphate (CS) glycosaminoglycans are sufficient to direct a selective inhibition of neurite growth from ventral temporal (VT) but not from dorsal nasal (DN) retina in mouse embryos; this may underlie the formation of axon divergence in the optic chiasm. Explants from the retinal region of embryonic day-14 mouse were grown on a laminin-polylysine substrate near to a circular spot coated with CS. In control cultures, in which no CS was added to the spot, both VT and DN retinal neurites grew extensively into the coated territory. When presented with spots coated with 10 mg/ml CS, neurite growth from the VT retina into the CS territory was dramatically reduced but that from the DN retina was not significantly affected. The selective inhibition to VT neurites was completely abolished by treatment with chondroitinase ABC, indicating a specific contribution of CS glycosaminoglycan in this regionally specific behaviour. This differential behaviour was not observed in explants presented with a lower or higher concentration of CS or in explants grown on substrate coated with a different laminin concentration. Thus, a critical ratio of CS to laminin seems to be essential to induce this differential behaviour in retinal neurites towards contact with CS. Furthermore, this behavior was not observed in explants cultured directly on a CS-rich substrate, suggesting that contact with growth-promoting molecules is necessary for the selective responses of retinal neurites during subsequent contact with CS. We concluded that CS glycosaminoglycan is sufficient to drive selective inhibition of VT but not DN neurites and that, together with a critical combination of growth promoting factors, it may control the axon divergence process at the mouse optic chiasm. PMID- 15902502 TI - Loss of ovarian function and the risk of ovarian cancer. AB - Animal models with premature ovarian failure resulting from the loss or depletion of germ cells consistently develop ovarian surface epithelial cell hyperplasia with invasion into the stroma and the development of ovarian tubular adenomas. In human ovaries, deep epithelial invaginations and inclusion cysts occur at increasing frequency with age and are thought to be the structures from which the majority of ovarian cancers arise. A feature that is common to these animal models and to post-menopausal women is a deficiency in the number of oocytes. The potential consequences of the loss or depletion of female germ cells, naturally or otherwise, include failure of follicle development, significant reductions in oestrogen and progesterone levels and elevation of circulating levels of gonadotropins. This review will consider the way in which these structural and hormonal changes affect ovarian cancer risk. Some lessons may be learned from gonad formation, since notable similarities exist between ovarian tumorigenesis and embryonic gonadogenesis including fragmentation of the basement membrane underlying the coelomic (surface) epithelium, the potential for the migration of epithelial cells into the gonad and the importance of the germ cells for the regulation of ovarian structure and function. PMID- 15902503 TI - Characterization of mussel gill cells in vivo and in vitro. AB - Mussel gill cells are attractive models in ecotoxicological studies because gills are the first uptake site for many toxicants in the aquatic environment; gill cells are thus often affected by exposure to pollutants. Our aim was to characterize mussel gill cells in vivo and in vitro by using morphological, histochemical and functional end-points. In paraffin sections stained with haematoxylin-eosin, three zones were distinguished in the long central gill filaments: frontal, intermediate and abfrontal. Various types of ciliated cells were present in the frontal zone, and both ciliated and non-ciliated cells were found in the abfrontal zone. The intermediate zone was comprised of flattened endothelial cells. Lipofuscin granules occurred in the three zones in variable amounts, depending on the specimen. Haemocytes were found in the haemolymph sinus of gill filaments. Mucocytes were identified in both frontal and abfrontal zones by means of periodic acid Schiff-alcian blue (PAS-AB) staining. In cryostat sections, succinate dehydrogenase (SDH) activity was mainly found in ciliated cells, whereas neutral lipids and acid-phosphatase-reactive lysosomes were present in all portions of the gill filament, mostly being related to lipofuscin granules. In mussels exposed to 5'-bromo-2'-deoxyuridine in vivo, proliferating cells were scattered throughout the gill filament. Gill cells (typically 2x10(7) cells/ml per mussel; 95% viability) were isolated by dissociation with dispase. Gill cell suspensions were heterogeneous: 58% were ciliated epithelial cells (positive for SDH), 42% were non-ciliated cells (including epithelial cells and haemocytes), 2.3% were mucocytes (positive for PAS-AB) and 4.25% were haemocytes (able to phagocytose neutral red-stained zymosan). Gill cell cultures were maintained up to 18 days without changing the culture medium, viability decreasing below 50% at day 18. Primary cultures of mussel gill cells might therefore be useful models for the in vitro assessment of xenobiotic impacts on coastal and estuarine ecosystems. PMID- 15902504 TI - Nuclear localization of the major vault protein in U373 cells. AB - The major vault protein (MVP) is the predominant member of a large ribonucleoprotein particle, named vault. Vaults are abundant in the cytosol of mammalian cells. Mammalian MVP has previously been reported to be associated with the nucleus, particularly its cytosolic surface on which vaults are thought to dock at or near the nuclear pore complex. To date the presence of vault particles inside the nucleus has been convincingly reported only for sea urchin cells. We have addressed the potential nuclear localization of MVP in mammalian cells by employing confocal laser microscopy and cryo-immunoelectron microscopy. As revealed by immunostaining and by analysis of cells transfected with a construct encoding MVP and green fluorescent protein, MVP is present in both the cytosol and in the nucleus. Cryo-electron microscopy of human astroglioma U373 cells reveals clusters of immunogold particles at nuclear pores and in the nucleoplasm suggesting that nuclear MVP is associated with particulate structures. Quantification of the fluorescence observed in the cytosol and in the nuclei reveals that about 5% of the MVP in U373 cells is localized inside the nucleus. Our results further support the notion that part of the cellular MVP can enter the nucleus. PMID- 15902505 TI - Subset of cells immunopositive for neurokinin-1 receptor identified as arterial interstitial cells of Cajal in human large arteries. AB - In the adventitia of large arteries, dendritic cells are located between nerve fibers, some of which contain substance P. The aim of the present study was to examine whether neurokinin 1 receptor (NK-1R) was expressed by dendritic cells in the arterial wall. Parallel sections of aortic and carotid artery segments were immunostained with anti-NK-1R and cell-type-specific antibodies. Dendritic cells in the arterial wall expressed NK-1R, albeit at a low level. Other cells, which intensely expressed NK-1R, were located along the border between the media and adventitia. They did not co-express any dendritic cell markers, including fascin, CD1a, S100, or Lag-antigen, and were negative for CD68, CD3, and mast cell tryptase. These NK-1R(+) cells were laser-capture microdissected and studied by means of electron-microscopic analysis. The microdissected cells were in direct contact with nerve endings, and their ultrastructure was typical of the interstitial cells of Cajal present in the gastrointestinal tract. Further systematic electron-microscopic analysis revealed that the cells displaying the features typical of interstitial cells of Cajal were a basic element of the human arterial wall architectonics. Arterial interstitial cells of Cajal were negative for c-kit but they expressed vasoactive intestinal peptide receptor 1 (VIPR1). Destructive alterations of contacts between arterial interstitial cells of Cajal and nerve endings were observed in arterial segments with atherosclerotic lesions. The functional significance of the arterial interstitial cells of Cajal and their possible involvement in atherosclerosis and other vascular diseases need clarification. PMID- 15902506 TI - Climatic controls of vegetation vigor in four contrasting forest types of India- evaluation from National Oceanic and Atmospheric Administration's Advanced Very High Resolution Radiometer datasets (1990-2000). AB - Ten-day advanced very high resolution radiometer images from 1990 to 2000 were used to examine spatial patterns in the normalized difference vegetation index (NDVI) and their relationships with climatic variables for four contrasting forest types in India. The NDVI signal has been extracted from homogeneous vegetation patches and has been found to be distinct for deciduous and evergreen forest types, although the mixed-deciduous signal was close to the deciduous ones. To examine the decadal response of the satellite-measured vegetation phenology to climate variability, seven different NDVI metrics were calculated using the 11-year NDVI data. Results suggested strong spatial variability in forest NDVI metrics. Among the forest types studied, wet evergreen forests of north-east India had highest mean NDVI (0.692) followed by evergreen forests of the Western Ghats (0.529), mixed deciduous forests (0.519) and finally dry deciduous forests (0.421). The sum of NDVI (SNDVI) and the time-integrated NDVI followed a similar pattern, although the values for mixed deciduous forests were closer to those for evergreen forests of the Western Ghats. Dry deciduous forests had higher values of inter-annual range (RNDVI) and low mean NDVI, also coinciding with a high SD and thus a high coefficient of variation (CV) in NDVI (CVNDVI). SNDVI has been found to be high for wet evergreen forests of north-east India, followed by evergreen forests of the Western Ghats, mixed deciduous forests and dry deciduous forests. Further, the maximum NDVI values of wet evergreen forests of north-east India (0.624) coincided with relatively high annual total precipitation (2,238.9 mm). The time lags had a strong influence in the correlation coefficients between annual total rainfall and NDVI. The correlation coefficients were found to be comparatively high (R2=0.635) for dry deciduous forests than for evergreen forests and mixed deciduous forests, when the precipitation data with a lag of 30 days was correlated against NDVI. Using multiple regression approach models were developed for individual forest types using 16 different climatic indices. A high proportion of the temporal variance (>90%) has been accounted for by three of the precipitation parameters (maximum precipitation, precipitation of the wettest quarter and driest quarter) and two of the temperature parameters (annual mean temperature and temperature of the coldest quarter) for mixed deciduous forests. Similarly, in the case of deciduous forests, four precipitation parameters and three temperature parameters explained nearly 83.6% of the variance. These results suggest differences in the relationship between NDVI and climatic variables based upon the time of growing season, time interval and climatic indices over which they were summed. These results have implications for forest cover mapping and monitoring in tropical regions of India. PMID- 15902507 TI - Contributing factors affecting the prognosis surgical outcome for thoracic OLF. AB - The thoracic ossification of ligamentum flavum (OLF) is a disease that produces spastic paraparesis, and there are various factors that may affect the surgical outcome of thoracic OLF patients. The authors of this study treated 19 of these thoracic OLF patients from 1998 to 2002, and retrospectively reviewed the patients' age, sex, symptom duration, involved disease level, preoperative clinical features, neurological findings, radiological findings, the other combined spinal diseases and the surgical outcomes. There were excellent or good surgical outcomes in 16 patients, but 3 patients did not improve after thoracic OLF surgery: this included 1 patient, whose motor function worsened after decompressive thoracic OLF surgery. The favorable contributing factors of surgical outcome in thoracic OLF are a short preoperative symptom duration, single-level lesion, and unilateral lesion type on CT axial scan. On the contrary, the poor prognostic factors are beak type lesion and intramedullary signal changes on T(2)-weighted sagittal MRI. The complete preoperative evaluation including radiologic findings will provide valuable aid in presuming the surgical outcome for the thoracic OLF patients. PMID- 15902508 TI - Root surface conditioning with nicotine or cotinine reduces viability and density of fibroblasts in vitro. AB - The purpose of study was to evaluate fibroblast attachment and cellular morphology on root surfaces chemically conditioned with nicotine or cotinine. A secondary objective was to determine if mechanical scaling and root planning of these chemically conditioned surfaces would alter cellular attachment. Root surface dentin specimens were prepared from uniradicular teeth of non-smoking patients. Specimens were randomly assigned to two experimental groups: no treatment (chemical conditioning only) and scaling and root planning after conditioning (SRPC). The concentrations of the tested substances were in the range of 0-1 mg/mL (nicotine) and 0-1 g/mL (cotinine). After a 24-h conditioning period, dentin slices were incubated with continuous lineage of fibroblastic cells from rat (McCoy cells) for another 24 h. Specimens were prepared for SEM analysis and microphotographs. The statistical analysis of the data indicated significant alteration of cellular morphology on fibroblasts that were grown on root surface exposed to nicotine concentrations greater than 1 ?g/mL. This effect of nicotine was not reduced by SRPC. On the other hand, in the SRPC group cellular density was greater. For cotinine-conditioned specimens, the greater concentrations also led to alteration on morphology, and these alterations were observed in the SRPC group as well. Cotinine did not induce significant changes on cellular density. The results indicated that fibroblasts are negatively influenced by nicotine present on the dentin substrate and also that scaling may reduce these effects. Cotinine treatment on root surfaces may alter cell morphology and density but these effects were less severe than that promoted by nicotine, and were not affected by scaling. PMID- 15902509 TI - L-Threonine dehydrogenase from the hyperthermophilic archaeon Pyrococcus horikoshii OT3: gene cloning and enzymatic characterization. AB - A gene encoding the L-threonine dehydrogenase homologue has been identified in a hyperthermophlic archaeon Pyrococcus horikoshii OT3 via genome sequencing. The gene was cloned and expressed in Escherichia coli. The purified enzyme from the recombinant E. coli was extremely thermostable; the activity was not lost after incubation at 100 degrees C for 20 min. The enzyme (molecular mass: 192 kDa) is composed of a tetrameric structure with a type of subunit (41 kDa). The enzyme is specific for NAD and utilizes L-threonine, L-serine and DL-threo-3-phenylserine as the substrate. The enzyme required divalent cations such as Zn(2+), Mn(2+) and Co(2+) for the activity, and contained one zinc ion/subunit. The K(m) values for L-threonine and NAD at 50 degrees C were 0.20 mM and 0.024 mM, respectively. Kinetic analyses indicated that the L-threonine oxidation reaction proceeds via a random mechanism with regard to the binding of L-threonine and NAD. The enzyme showed pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of NADH. This is the first description of the characteristics of an L-threonine dehydrogenase from the archaea domain. PMID- 15902510 TI - How to be moderately halophilic with broad salt tolerance: clues from the genome of Chromohalobacter salexigens. AB - We analyzed the amino acid composition of different categories of proteins of the moderately halophilic bacterium Chromohalobacter salexigens, as deduced from its genome sequence. Comparison with non-halophilic representatives of the gamma Proteobacteria (Escherichia coli, Pseudomonas aeruginosa, Vibrio cholerae) shows only a slight excess of acidic residues in the cytoplasmic proteins, and no significant differences were found in the acidity of membrane-bound proteins. In contrast, a very pronounced difference in mean pI value was observed for the periplasmic binding proteins of the ABC transport systems of C. salexigens and the non-halophiles E. coli and P. aeruginosa. V. cholerae, which is adapted to life in brackish water, showed intermediate values. The findings suggest that there is a major difference between the proteins of the moderate halophile C. salexigens and non-halophilic bacteria in their periplasmic proteins, exemplified by the substrate binding proteins of transport systems. The highly acidic nature of these proteins may enable them to function at high salt concentrations. The evolution of highly salt-tolerant prokaryotes may have depended on an increase in acidity of the proteins located external to the cytoplasmic membrane, enabling effective transport of nutrients into the cell. PMID- 15902511 TI - Incisional hernia--comparison of mesh repair with Cardiff repair: an university hospital experience. AB - BACKGROUND: Incisional hernia is a frequent complication of abdominal surgery. Various types of repair are recommended for incisional hernia. Suture and mesh repair are compared in the present study. METHOD: One hundred seventy one patients with incisional hernia underwent Cardiff repair (far and near sutures with reinforcement sutures) which was used as an open suture repair while onlay polypropylene mesh was used in the mesh repair technique. RESULT: Cardiff repair was performed in 116 patients with no mortality with recurrence in two patients with mean follow up of 7.1 years. Both these patients with recurrence had a defect measuring more than 10 cm in width. Mesh repair was carried out in 55 patients with no recurrence in mean follow up of 37 months. Seroma formation was noted in 7 (12.72%) with mesh repair as compared to 4 (3.44%) patients with Cardiff repair. CONCLUSION: We recommend Cardiff repair for primary and small to medium size incisional hernias. Onlay polypropylene mesh is ideal for tension free hernia repair, recurrent incisional hernia and hernia defects wider than 10 cm. PMID- 15902512 TI - The relationship between C677T methylenetetrahydrofolate reductase gene polymorphism and retinopathy in type 2 diabetes: a meta-analysis. AB - The association between retinopathy in type 2 diabetes [diabetic retinopathy (DR)] and the C677T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene has been investigated in several case-control studies. These studies rendered contradictory results, some indicating that the polymorphism is associated with the risk of developing DR whereas others concluded there is no association. To shed light on these inconclusive findings, a meta-analysis of all available studies relating the C677T polymorphism to the risk of developing DR was conducted. Four out of five identified studies included populations of East Asian descent, and only one involved samples from European descent (Caucasians). Overall, the meta-analysis suggested large heterogeneity between studies (p = 0.08, I(2) = 52%) and marginal association between C677T transition and the risk of developing DR: random effects odds ratio (OR) = 1.39 [95% CI (1.05, 1.83)]. The sensitivity analysis [exclusion of one East Asian study with the controls not in Hardy-Weinberg equilibrium (HWE)] showed no heterogeneity (p = 0.25, I(2) = 27%) and no significant association: fixed effects OR = 1.22 [95% CI (0.99, 1.51)] and random effects OR = 1.24 [95% CI (0.96, 1.60)]. The sub-group analysis for the East Asian population produced a significant association: fixed effects OR = 1.48 [95% CI (1.20, 1.83)] and random effects OR = 1.52 [95% CI (1.14, 2.03)]. However, sensitivity analysis in East Asians revealed that the association is marginal: fixed effects OR = 1.33 [95% CI (1.04, 1.70)] and random effects OR = 1.36 [95% CI (1.01, 1.83)]. There is a source of bias in the selected studies: the largest studies failed to show association while the smallest study claimed an association. The above findings reinforce the need for larger and more rigourous studies in this area. PMID- 15902513 TI - A case of tuberculous meningoencephalitis in a patient with Behcet's disease. PMID- 15902514 TI - Calcinosis cutis universalis in a patient with systemic lupus erythematosus. AB - Deposition of calcium salts in the skin and subcutaneous tissue occurs in a variety of rheumatic diseases, being most commonly associated with scleroderma, CREST (calcinosis, Raynaud's phenomenon, esophageal dysfunction, sclerodactyly, and telangiectasia), dermatomyositis, and overlap syndromes but is a rare complication of systemic lupus erythematosus (SLE). Calcinosis is classified into four subsets: dystrophic, metastatic, idiopathic, or calciphylaxis/iatrogenic. The pathophysiology of calcinosis cutis remains unclear. Our patient developed extensive areas of calcifications in the trunk and extremities (calcinosis universalis) 8 years after SLE diagnosis, which would correspond to a form of dystrophic calcification. No response was observed after treatment with oral diltiazem for 3 months. We review the literature on the pathogenesis and prevalence of calcinosis universalis in SLE. PMID- 15902515 TI - Measuring utilities by the time trade-off method in Tunisian rheumatoid arthritis patients. AB - The objective of this study was to determine the feasibility, reliability and validity of the time trade-off (TTO) in Tunisian rheumatoid arthritis (RA) patients. The TTO was used to measure the utility in 122 RA patients with increasing difficulty in performing activities of daily living. The 1-week test retest reproducibility was studied in 57 patients using the intraclass correlation coefficient (ICC). Validity was evaluated by comparison with other outcome measures: utility rating scale (RS), quality of life (QOL) [arthritis impact measurement scale 2 (AIMS2), rheumatoid arthritis quality of life (RAQOL)], functional status [health assessment questionnaire (HAQ), Lee index] and disease activity score (DAS). Eight patients (6.6%) did not complete the TTO. The median value of the TTO score was 0.655 (0.019-1.000). The ICC for reliability of the TTO was 0.89 (p<0.001). The TTO showed poor to moderate correlation (Spearman's correlation coefficients between 0.2 and 0.409, p<0.01) with AIMS2, RAQOL, HAQ and Lee index. We did not find any correlation between TTO and DAS. Multiple regression analysis showed that only 32% of TTO scores could be explained. The TTO method appeared to be reliable in a group of Tunisian RA patients, but TTO values were poorly to moderately related to measures of QOL, functional ability, and disease activity. We think that TTO and RS are not feasible for use in RA patients. PMID- 15902516 TI - Perinuclear antineutrophil cytoplasmic antibody myeloperoxidase-positive vasculitis in association with ulcerative colitis. AB - We describe a patient with ulcerative colitis (UC) who developed small vessel vasculitis. Perinuclear antineutrophil cytoplasmic antibody myeloperoxidase (p ANCA-MPO) positivity was detected along with a highly elevated titer of anticardiolipin antibodies. A total proctocolectomy was undertaken and the patient, more than 5 years later, remains in very good condition. The possible causative association between the UC, the p-ANCA-MPO-positive small vessel vasculitis, and the anticardiolipin antibodies is discussed. PMID- 15902517 TI - Fibromyalgia and headache: an epidemiological study supporting migraine as part of the fibromyalgia syndrome. AB - Fibromyalgia is defined by widespread body pain, tenderness to palpation of tender point areas, and constitutional symptoms. The literature reports headache in about half of fibromyalgia patients. The current epidemiological study was designed to determine the prevalence and characteristics of headache in fibromyalgia patients. Treatment-seeking fibromyalgia patients were evaluated with measures for fibromyalgia, chronic headache, quality of life, and psychological distress. Multivariate analysis of variance (MANOVA) and t-tests were used to identify significant differences, as appropriate. A total of 100 fibromyalgia patients were screened (24 fibromyalgia without headache and 76 fibromyalgia with headache). International Headache Society diagnoses included: migraine alone (n = 15 with aura, n = 17 without aura), tension-type alone (n = 18), combined migraine and tension-type (n = 16), post-traumatic (n = 4), and probable analgesic overuse headache (n = 6). Fibromyalgia tender point scores and counts and most measures of pain severity, sleep disruption, or psychological distress were not significantly different between fibromyalgia patients with and without headache. As expected, the fibromyalgia patients with headache scored higher on the Headache Impact Test (HIT-6) (62.1 +/- 0.9 vs 48.3 +/- 1.6, p < 0.001). HIT-6 scores were >60 in 80% of fibromyalgia plus headache patients, representing severe impact from headache, and 56-58 in 4%, representing substantial impact. In summary, chronic headache was endorsed by 76% of treatment seeking fibromyalgia patients, with 84% reporting substantial or severe impact from their headaches. Migraine was diagnosed in 63% of fibromyalgia plus headache patients, with probable analgesic overuse headache in only 8%. General measures of pain, pain-related disability, sleep quality, and psychological distress were similar in fibromyalgia patients with and without headache. Therefore, fibromyalgia patients with headache do not appear to represent a significantly different subgroup compared to fibromyalgia patients without headache. The high prevalence and significant impact associated with chronic headache in fibromyalgia patients, however, warrants inclusion of a headache assessment as part of the routine evaluation of fibromyalgia patients. PMID- 15902518 TI - Concurrent occurrence of Sweet's syndrome and erythema nodosum: an overlap in the spectrum of reactive dermatoses. AB - Simultaneous occurrence of Sweet's syndrome and erythema nodosum is very rare. We describe a case of a young male with a recent history of streptococcal infection who presented with concurrent Sweet's syndrome and erythema nodosum. Although the exact pathogenesis of these dermatoses is not yet clear, their similarities and simultaneous occurrence suggest a possible common underlying mechanism and may represent a continuum of reactive dermatoses. Evaluation of the role of cytokines in the etiopathogenesis of these conditions will be useful for further assessment and treatment of these conditions. Like the association of acanthosis nigricans and certain cancers and diabetes, Sweet's syndrome and erythema nodosum may be associated with certain malignancies, autoimmune disorders, or inflammatory bowel disease. Early recognition of these skin lesions can guide a search for underlying disorders. Patients with Sweet's syndrome should undergo an age appropriate work-up for malignancy. PMID- 15902519 TI - Infliximab as monotherapy in giant cell arteritis. PMID- 15902520 TI - Acute pseudogout following contrast angiography. PMID- 15902521 TI - Arterial occlusion in systemic lupus erythematosus: a good prognostic sign? AB - Arterial occlusion with subsequent amputation of extremities is a rare manifestation of systemic lupus erythematosus (SLE). It may be caused by local arteritis and/or thrombosis. We describe the clinical and laboratory manifestations and treatment administered to six SLE patients who developed peripheral arterial necrosis necessitating amputation of extremities secondary to the arterial occlusion. All patients were female, with ages ranging from 16 to 65 years. Arterial occlusion took place in the initial months of disease (median: 7 months). Only one of five patients tested for antiphospholipid antibodies had these antibodies who also had vasculitis and thrombosis in a histopathological study. Most patients presented a very benign outcome after the amputation of extremities and stayed in remission for several years. The satisfactory outcome of most patients after the vascular phenomenon allows us to consider the possibility that such a complication could be, for unknown reasons, a marker for good prognosis in SLE or, alternatively, that the aggressive therapy administrated for patients with this complication at the beginning of the disease could recover the balance of the immune system, avoiding future relapses. PMID- 15902522 TI - Successful treatment of rapidly progressive interstitial pneumonia with autologous peripheral blood stem cell transplantation in a patient with dermatomyositis. AB - Aggressive autoimmune diseases are often treated by intensive immunosuppressive treatment such as high-dose methylprednisolone and intravenous cyclophosphamide. Autologous hematopoietic stem cell transplantation can facilitate high-dose immunosuppressive therapy (HDIT), which is myeloablative. We describe a 54-year old female patient with rapidly progressive and refractory interstitial pneumonia due to dermatomyositis, which was successfully treated with high-dose cyclophosphamide and autologous blood stem cell transplantation. Following transplantation, dyspnea disappeared, and arterial blood gas analysis and respiratory function test showed marked improvement. This improvement was confirmed by diminished interstitial shadows on chest X-ray and computed tomography scans. Eighteen months after transplantation, the patient is doing well without symptoms and signs of interstitial pneumonia. PMID- 15902523 TI - Predictors of outcome in surgical treatment for basal joint osteoarthritis of the thumb. AB - In this series, several preoperative factors influencing the outcome in surgically treated patients with carpometacarpal osteoarthritis of the thumb were analyzed. Thirty-six patients were studied. Hand dominance, interphalangeal joint motion, radiographic stage, and surgical procedure did not reach significance. In contrast, age, first web retraction, and hyperextension of the metacarpophalangeal joint were considered valuable in predicting the ultimate outcome. PMID- 15902524 TI - Trapezius muscle imbalance in individuals suffering from frozen shoulder syndrome. AB - This aim of this study was to characterize upper and lower trapezius muscle activity for patients experiencing frozen shoulder syndrome (FSS) compared to asymptomatic subjects. Fifteen patients suffering from unilateral FSS and 15 asymptomatic subjects voluntarily participated in this study. Data were gathered on electromyographic (EMG) activity obtained from the upper and lower trapezius muscles during maximal static arm elevations at six different testing positions: 60 and 120 degrees of flexion, abduction in the frontal plane, and abduction in the scapular plane. The group with FSS revealed increased upper trapezius EMG activity at the 60 degrees (mean difference = 12%, p < 0.003) and 120 degrees (mean difference = 24%, p < 0.004) testing positions, and increased lower trapezius EMG activity at the 120 degrees testing positions (mean difference = 6%, p < 0.002), compared to asymptomatic subjects. Higher ratios of the upper trapezius to lower trapezius EMG activity were also found in the patient group (p < 0.0005) compared to asymptomatic subjects. The results of this study indicate that the increased trapezius muscle activity may contribute to scapular substitution movement in compensation for impaired glenohumeral motion in patients with FSS. The insufficiency of the increased lower trapezius muscle activity should be an important consideration in the rehabilitation of patients experiencing FSS. PMID- 15902525 TI - Non-rheumatoid erosive arthritis associated with type I hereditary angioedema. AB - Hereditary angioedema (HAE) is an autosomal dominant disease that causes recurrent attacks of non-pitting edema of soft tissues, without pruritus. This disorder can also affect internal organs. The cause of HAE consists in quantitative or qualitative defective production of C1 inhibitor (C1-INH). Many autoimmune diseases such as systemic lupus erythematosus (SLE) (or SLE-like syndromes), Sjogren's syndrome, scleroderma, thyroiditis, glomerulonephritis, and inflammatory bowel disease have been described in patients suffering from HAE. A concomitance with pure arthritis was previously reported only in two adult patients. Here, we describe for the first time the association between HAE and a non-rheumatoid erosive oligoarthritis involving hips and wrists. PMID- 15902526 TI - Risedronate-induced intravascular haemolysis complicated by acute tubular necrosis. PMID- 15902527 TI - Giant cell arteritis in a patient with acute aortic insufficiency with thyrotoxicosis. AB - Acute aortic insufficiency in the setting of thyrotoxicosis can mask the presentation of vasculitis. We report a case of a 38-year-old woman with a 22 weeks gestation pregnancy who was known to be hyperthyroid for 4 months prior to conception. She presented with thyrotoxicosis and acute respiratory failure. Echocardiogram revealed severe acute aortic regurgitant flow. Following medical treatment for aortic insufficiency and thyrotoxicosis, the patient underwent ascending aorta replacement with aortic valve repair. Pathological exam revealed giant cell arteritis. Both giant cell arteritis and thyrotoxicosis share a common major histocompatibility antigen which may facilitate concomitant disease presentation. Following immunosuppression for giant cell arteritis, valve repair, and treatment for thyrotoxicosis, the patient made a complete recovery. A rise in human chorionic gonadotropin (HCG) during the first trimester of pregnancy is known to have a stimulatory effect on the thyroid gland and may result in hyperthyroidism. Although HCG may have exacerbated the existing hyperthyroidism, in this case it was not causal, as the diagnosis preceded her pregnancy by several months. Diagnosis of vasculitis may be overshadowed by the presence of thyrotoxicosis. Significant vascular compromise in the setting of thyrotoxicosis must prompt an evaluation for vasculitis. This may prevent unnecessary surgery with attendant morbidity and mortality. PMID- 15902528 TI - Synovial nitric oxide concentrations are increased and correlated with serum levels in patients with active Behcet's disease: a pilot study. AB - Behcet's disease (BD) is a relapsing immunoinflammatory vasculitis of unknown etiology characterized by endothelial dysfunction. Articular symptoms and signs are present in about 75% of cases and characterized by seronegative arthritis and nonspecific synovitis. We demonstrated that both serum and erythrocyte nitric oxide (NO(.)) levels, the most abundant free radical in the body, were elevated in BD and associated with disease activity. This study further investigated NO(.) levels in the synovial fluid and serum from patients with active and inactive BD. A total of 23 BD patients with articular involvement (14 men and 9 women) satisfying International Study Group criteria and 15 age- and sex-matched healthy control subjects (9 men and 6 women) undergoing elective arthroscopy were included in this case-control investigation. The synovial fluid and serum were obtained from BD patients and controls. Clinical and laboratory findings including neutrophil count and erythrocyte sedimentation rate (ESR) were used to classify BD patients as active (n = 11) or inactive (n = 12). Synovial as well as serum NO(.) levels were compared between the groups and correlation analysis was performed. Acute phase reactant levels were significantly higher (for each, p < 0.01) in BD patients than control subjects in the active period. The mean synovial NO(.) level in active Behcet's patients (mean +/- SD 76.61 +/- 11.95 micromol/l) was significantly higher than in inactive patients (46.16 +/- 8.89 micromol/l, p < 0.001) and healthy control subjects (39.60 +/-8.03 micromol/l, p < 0.001). The difference between inactive patients and controls was not significant (p > 0.05). Active BD patients had significantly higher serum NO(.) levels (38.84 +/- 9.15 micromol/l) than inactive patients (30.91 +/- 5.88 micromol/l, p = 0.018) and control subjects (28.86 +/- 5.91 micromol/l, p = 0.002). In addition, synovial NO(.) levels were positively correlated with serum levels (r(2) = 0.621, p < 0.001). Increased synovial NO(.) levels in active BD patients probably reflect a nonspecific inflammatory process of the synovium and, therefore, arthralgia and arthritis as a common finding of BD. PMID- 15902529 TI - Value of intensive diagnostic microbiological investigation in low- and high-risk patients with community-acquired pneumonia. AB - In a prospective study to evaluate the diagnostic yield of different microbiological tests in hospitalised patients with community-acquired pneumonia, material for microbiological investigation was obtained from 262 patients. Clinical samples consisted of the following: sputum for Gram staining, culture, and detection of pneumococcal antigen; blood for culture and serological tests; urine for detection of Legionella pneumophila serogroup 1 antigen and pneumococcal antigen; and specimens obtained by fiberoptic bronchoscopy. A pathogen was identified in 158 (60%) patients, with Streptococcus pneumoniae (n=97) being the most common causative agent of community-acquired pneumonia. In 82% of the 44 patients with an adequate sputum specimen, a positive Gram stain was confirmed by positive sputum culture. S. pneumoniae infections were detected principally when adequate sputum specimens were examined by Gram stain and culture and when adequate and inadequate sputum specimens were tested for the presence of pneumococcal antigen (n=58; 60%). The urinary pneumococcal antigen test was the most valuable single test for detection of S. pneumoniae infections (n=52; 54%) when sputum pneumococcal antigen determination was not performed. Fiberoptic bronchoscopy was of additive diagnostic value in 49% of the patients who did not expectorate sputum and in 52% of those in whom treatment failed. Investigation of sputum by a combination of Gram stain, culture, and detection of pneumococcal antigen was the most useful means of establishing an aetiological diagnosis of community-acquired pneumonia, followed by testing of urine for pneumococcal antigen. Fiberoptic bronchoscopy may be of additional value when treatment failure occurs. PMID- 15902530 TI - Clinical characteristics and significance of Streptococcus salivarius bacteremia and Streptococcus bovis bacteremia: a prospective 16-year study. AB - The aim of this study was to determine the clinical significance of Streptococcus salivarius isolates recovered from blood cultures and compare them with isolates of Streptococcus bovis biotypes I and II. Seventeen of the 52 (32%) S. salivarius isolates recovered were considered clinically significant, compared with 62 of the 64 (97%) S. bovis isolates (p<0.0001). Bacteremia caused by S. salivarius occurred mostly in patients who showed relevant disruption of the mucous membranes and/or serious underlying diseases. Patients with S. salivarius bacteremia were younger than those with S. bovis bacteremia (57 vs. 67 years; p<0.01). Patients with S. salivarius bacteremia and patients with S. bovis II bacteremia had similar rates of endocarditis, colon tumors, and non-colon cancer. On the other hand, when compared with S. bovis I bacteremia, S. salivarius bacteremia was associated with lower rates of endocarditis (18% vs. 74%, respectively) (p<0.01) and colon tumors (0% vs. 57%, respectively) (p<0.005) and higher rates of non-colon cancer (53% vs. 9.5%, respectively) (p<0.01). Bacteremia caused by S. bovis II had a hepatobiliary origin in 50% of the patients, while, in contrast, that due to S. salivarius or S. bovis I was less frequently associated with a hepatobiliary origin (12% and 5%, respectively) (p<0.00001). The rate of penicillin resistance was 31% among S. salivarius isolates and 0% among S. bovis isolates (p<0.0001). In conclusion, the clinical characteristics of S. salivarius bacteremia and S. bovis II bacteremia are similar, and the isolation of S. salivarius in blood should not be systematically regarded as contamination. PMID- 15902531 TI - Risk factors for infections with Norovirus gastrointestinal illness in Switzerland. AB - Viral infections, particularly those caused by noroviruses (NV, genus Norovirus), are the most common cause of community-acquired gastroenteritis in Europe, with respect to both endemic and epidemic occurrence. For the first time, a general practitioner-based case-control study was performed between July 2001 and July 2003 in the German-speaking part of Switzerland in order to identify risk factors for sporadic NV infections. The consumption of different foodstuffs and of bottled mineral water did not show any significant association with the risk of NV gastroenteritis, nor was there any significant effect of individual ABO histo blood group or household size on the incidence of NV gastroenteritis. The findings are consistent with person-to-person transmission as the most important route of transmission for community-acquired, sporadic NV infection, in that 39% of all patients reported they had had contact with ill persons before their illness. The fact that 33% reported contact with ill persons, mainly within family groups, after their own illness suggested that a substantial proportion of patients were part of family mini-outbreaks. PMID- 15902532 TI - Clonal analysis of invasive pneumococcal isolates in Scotland and coverage of serotypes by the licensed conjugate polysaccharide pneumococcal vaccine: possible implications for UK vaccine policy. AB - A 7-valent pneumococcal conjugate vaccine (PCV7) has gained licensure and has proven successful in the USA for preventing pneumococcal disease and reducing the incidence of antibiotic-resistant pneumococcal strains. The ability, therefore, to accurately monitor the likely effect of the introduction of PCV7 vaccine on invasive pneumococcal disease in the UK is essential. Serotyping and multilocus sequence typing was performed on invasive isolates of Streptococcus pneumoniae (n=645) from Scotland during 2003. The information gained from this was used to evaluate serotype coverage by the vaccine and the relationship between serotypes. In the present study, invasive pneumococcal disease in Scotland was caused by 33 different serotypes, consisting of 150 sequence types. Overall, 48.4% of the isolates were of serotypes included in the PCV7. Pneumococci were most frequently associated with sequence types 9, 124, and 162. PCV7 would provide protection in 71.8% of infants under 5 years of age against the serotypes in the vaccine. There was limited evidence of the potential for capsule switch among currently circulating invasive pneumococci. The successful implementation of a suitable vaccination programme should lead to a reduction in invasive pneumococcal disease in the UK as well as a reduction in antibiotic resistance of pneumococcal strains. PMID- 15902533 TI - Importance of environmental transmission in cases of EHEC O157 causing hemolytic uremic syndrome. AB - A local outbreak of Shiga toxin (Stx)-producing enterohemorrhagic Escherichia coli (EHEC) O157:H7 causing severe hemolytic-uremic syndrome (HUS) was found to be caused by environmental transmission. Automated ribotyping and pulsed-field gel electrophoresis revealed that four stx2-positive EHEC isolates obtained from two unrelated children, one mother and one cow were identical. Results of an epidemiological investigation strongly suggest that both children were infected via a meadow strewn with manure containing EHEC-positive feces from the infected cow a few days prior to the onset of illness. The cow belonged to a cattle farm neighboring the meadow. This report highlights the risk of acquiring EHEC O157 through indirect contact with a farm environment. PMID- 15902534 TI - Value of measuring serum procalcitonin, C-reactive protein, and mannan antigens to distinguish fungal from bacterial infections. AB - The study presented here was conducted to determine the diagnostic value of measuring procalcitonin, C-reactive protein, and mannan antigens to distinguish fungal from bacterial infections. The sensitivity and specificity of these measurements ranged from 35% to 97%. On days 1 and 3 following the onset of fever, both serum procalcitonin and C-reactive protein levels were lower in patients with fungal infections than in those with bacterial infections (p<0.0001). The presence of mannan antigens combined with a procalcitonin level <0.5 ng/ml provided higher specificity for distinguishing fungal from bacterial infections than each result alone. PMID- 15902535 TI - In vitro activity of fosfomycin alone and in combination with amoxicillin, clarithromycin and metronidazole against Helicobacter pylori compared with combined clarithromycin and metronidazole. AB - In order to evaluate the suitability of fosfomycin in combination with other agents for the treatment of Helicobacter pylori infections, the susceptibility profiles of 65 H. pylori strains were determined against multiple antimicrobial agents and combinations thereof using the agar dilution method. For fosfomycin alone, the range of minimum inhibitory concentration (MIC) results and the MICs at which 50% and 90% of strains were inhibited were 0.5-32 microg/ml and 2 and 4 microg/ml, respectively. For the combination of fosfomycin with amoxicillin, clarithromycin or metronidazole, the means calculated for the minimum and maximum fractional inhibitory concentration index were 0.70-1.17 and 1.15-2.03, respectively, suggesting partial synergy or indifference in the majority of strains. The combination of clarithromycin and metronidazole showed synergistic activity against 14 of 28 H. pylori strains tested. The in vitro activity results suggest the combination of fosfomycin with either amoxicillin or clarithromycin may be a promising alternative for the treatment of H. pylori infection. However, the clinical efficacy of these regimens remains to be investigated. PMID- 15902536 TI - Phenotypic and genotypic characterization of prolyliminopeptidase-negative Neisseria gonorrhoeae isolates in Denmark. AB - In the study presented here 26 recent Danish clinical isolates of prolyliminopeptidase (PIP)-negative Neisseria gonorrhoeae were phenotypically and genotypically characterized to investigate whether one or more PIP-negative strains are circulating in the Danish community. The profiles of these isolates were compared with those of three isolates from a recent outbreak of PIP-negative N. gonorrhoeae infection in the UK. Twenty-five of the Danish isolates and all three UK isolates had similar antibiograms and were designated serovar IB-4. Genotypic characterization by pulsed-field gel electrophoresis, porB1b gene sequencing, and opa-typing revealed that these isolates were indistinguishable or closely related. The results indicate that at least one PIP-negative N. gonorrhoeae strain is currently circulating in the Danish community, and this strain is indistinguishable from the one that caused an outbreak in the UK. PMID- 15902537 TI - Occurrence and regional distribution of SHV-type extended-spectrum beta lactamases in Hungary. AB - In order to determine the presence and geographical distribution of SHV-type extended-spectrum beta-lactamase genes among Enterobacteriaceae strains in Hungary, isolates from 25 microbiology laboratories throughout the country were collected between January 2002 and August 2003 and examined. Sequencing of the genes showed that SHV-5 and SHV-2a are the dominant SHV-types in extended spectrum beta-lactamase-producing Enterobacteriaceae strains in this country. The SHV-2 gene, which is prevalent in many European countries, was not detected, but one isolate carried the SHV-12 gene. The results show that these genes are circulating among Enterobacteriaceae strains in Hungary and indicate that strict infection control measures are warranted in order to prevent their spread. PMID- 15902538 TI - Infected cat-bite wound treated successfully with moxifloxacin after failure of parenteral cefuroxime and ciprofloxacin. PMID- 15902539 TI - Usefulness of antibiotic-lock technique in management of oncology patients with uncomplicated bacteremia related to tunneled catheters. PMID- 15902540 TI - Pneumocystis pneumonia in an alcoholic patient with prolonged mechanical ventilation. PMID- 15902541 TI - First isolation and genotypic identification of Rickettsia conorii Malish 7 from a patient in Greece. PMID- 15902542 TI - Laboratory surveillance of influenza in northern Greece, 1993-2003. PMID- 15902543 TI - Complete mitochondrial DNA sequences of the decapod crustaceans Pseudocarcinus gigas (Menippidae) and Macrobrachium rosenbergii (Palaemonidae). AB - The complete mitochondrial DNA sequence was determined for the Australian giant crab Pseudocarcinns gigas (Crustacea: Decapoda: Menippidae) and the giant freshwater shrimp Macrobrachium rosenbergii (Crustacea: Decapoda: Palaemonidae). The Pse gigas and Mrosenbergii mitochondrial genomes are circular molecules, 15,515 and 15,772 bp in length, respectively, and have the same gene composition as found in other metazoans. The gene arrangement of M. rosenbergii corresponds with that of the presumed ancestral arthropod gene order, represented by Limulus polyphemus, except for the position of the tRNALeu(UUR) gene. The Pse. gigas gene arrangement corresponds exactly with that reported for another brachyuran, Portunus trituberculatus, and differs from the M. rosenbergii gene order by only the position of the tRNAHis gene. Given the relative positions of intergenic nonoding nucleotides, the "duplication/random loss" model appears to be the most plausible mechanism for the translocation of this gene. These data represent the first caridean and only the second brachyuran complete mtDNA sequences, and a source of information that will facilitate surveys of intraspecific variation within these commercially important decapod species. PMID- 15902544 TI - Physical molecular maps of wheat chromosomes. AB - In bread wheat, a set of 527 simple sequence repeats (SSRs) were tried on 164 deletion lines, leading to a successful mapping of 270 SSRs on 313 loci covering all 21 chromosomes. A maximum of 119 loci (38%) were located on B subgenome, and a minimum of 90 loci (29%) mapped on D subgenome. Similarly, homoeologous group 7 carried a maximum of 61 loci (19%), and group 4 carried a minimum of 22 loci (7%). Of the cited 270 SSRs, 39 had multiple loci, but only eight of these detected homoeologous loci. Linear order of loci in physical maps largely corresponded with those in the genetic maps. Apparently, distances between each of only 26 pairs of loci significantly differed from the corresponding distances on genetic maps. Some loci, which were genetically mapped close to the centromere, were physically located distally, while other loci that were mapped distally in the genetic maps were located in the proximal bins in the physical maps. This suggested that although the linear order of the loci was largely conserved, variation does exist between genetic and physical distances. PMID- 15902546 TI - Using competence network collaboration and decision-analytic modeling to assess the cost-effectiveness of interferon alpha-2b plus ribavirin as initial treatment of chronic hepatitis C in Germany. AB - The objective of this study was to translate and apply a decision-analytic model for chronic hepatitis C (CHC) to the German health care context using competence network collaboration. The German Hepatitis C Model (GEHMO) competence network used a systematic multistep approach to identify and transfer a high quality Markov model for CHC to the German health care context. GEHMO was used to project lifetime clinical and economic outcomes and to determine the cost-effectiveness of initial antiviral therapy with interferon a-2b plus ribavirin from a societal perspective. In 40-year-old patients combination therapy for 24 and 48 weeks increased life expectancy by 1.6 and 2.3 years, respectively, compared with interferon alone for 48 weeks. The discounted incremental cost-utility ratios (ICUR) for combination therapy were euro 5,500 per quality-adjusted life-year gained (QALY) for 24 weeks and euro 6,800/QALY for 48 weeks of treatment. ICUR was euro 9,800/QALY for moving from 24 to 48 weeks of treatment. Combination therapy remained cost-effective in sensitivity analyses. In conclusion, combination therapy with interferon alpha-2b and ribavirin is effective and cost effective compared with other well-accepted medical treatments. Competence network collaboration and decision modeling provide a useful and efficient approach to combine evidence from international studies with country-specific parameters. PMID- 15902547 TI - Frontal lobe-dependent functions in treated phenylketonuria: blood phenylalanine concentrations and long-term deficits in adolescents and young adults. AB - Even early-treated phenylketonurics may suffer from phenylalanine-related deficits. Elevated phenylalanine concentrations can interfere with the development and function of the CNS. Outcome beyond childhood has not been extensively investigated. This long-term study was performed to determine whether adolescents and young adults with PKU show frontal lobe-dependent deficits when compared to diabetic patients. The comparative study covered 35 PKU patients, 13 21 years of age (mean 17.8 years), and 35 diabetic patients matched for sex, age and socioeconomic status. Patients were assessed for IQ (Culture Fair Intelligence Test), information processing (Trail Making Test), and selective and sustained attention (Stroop Task, Test d-2). Assessments were repeated within a 3 year follow-up. PKU patients showed no increase in blood phenylalanine concentrations at follow-up. They had significantly poorer test results than the diabetic patients at both assessment times. Within the tests, however, this was due to reduced performance speed but not to deficits in specific frontal lobe dependent functions. Elevated phenylalanine concentrations seem to exert a global effect slowing performance speed. This effect is enduring in adolescence and early adulthood. PMID- 15902548 TI - Breastfeeding experience in inborn errors of metabolism other than phenylketonuria. AB - Breastfeeding has been recommended for the dietary treatment of infants with phenylketonuria, but studies documenting clinical experience in other inborn errors of metabolism are very few. Seven infants diagnosed with methylmalonyl-CoA mutase deficiency (n=2), ornithine carbamoyltransferase deficiency (n=1), propionic acidaemia (n=1), isovaleric acidaemia (n=1), maple syrup urine disease (n=1) and glutaric acidemia type I (n=1) were tried with breastfeeding over two years. After the control of acute metabolic problems, an initial feeding period with a measured volume of expressed breast milk plus a special essential amino acid mixture was continued with breastfeeding on demand and with the addition of a special essential amino acid mixture. Two patients with methylmalonic acidaemia and one patient with glutaric acidaemia type I tolerated breastfeeding on demand very well, with good growth and metabolic control for periods of 18, 8 and 5 months, respectively. In the patient with propionic acidaemia, on-demand breastfeeding continued for 3 months but was terminated after two acute metabolic episodes. The patient with isovaleric acidaemia had insufficiency of breast milk and formula supplementation ended with breast milk cessation. In the patient with severe ornithine carbamoyltransferase deficiency, breastfeeding was stopped owing to poor metabolic control. The patient with maple syrup urine disease also experienced problems, both in metabolic control and in insufficiency of breast milk, resulting in termination of breastfeeding. Breastfeeding of infants with inborn errors of protein catabolism is feasible, but it needs close monitoring with attention to such clinical parameters as growth, development and biochemistry, including amino acids, organic acids and ammonia. PMID- 15902549 TI - Elevated plasma citrulline and arginine due to consumption of Citrullus vulgaris (watermelon). AB - A 19-month-old girl with developmental delay was found to have moderately elevated plasma citrulline and mildly elevated plasma arginine concentrations. Dietary history revealed that she consumed large quantities of watermelon (Citrullus vulgaris), a fruit containing high free citrulline and arginine concentrations. In order to determine whether the patient's high watermelon intake could account for her elevated plasma citrulline and arginine concentrations, we studied the response of plasma citrulline and arginine to ingestion of watermelon in six healthy adult volunteers. All developed markedly elevated plasma citrulline (mean maximum 593 micromol/L, range 386-1069) and moderately elevated plasma arginine (mean maximum 199 micromol/L, range 128-251). Physicians and laboratory personnel performing metabolic investigations should be aware of watermelon-induced citrullinaemia. Its hallmarks are elevated plasma citrulline, and to a lesser extent arginine, in the absence of orotic or arginosuccinic aciduria or hyperammonaemia. This phenomenon has implications for the management of patients with urea cycle and related disorders. PMID- 15902550 TI - A survey of Japanese patients with Menkes disease from 1990 to 2003: incidence and early signs before typical symptomatic onset, pointing the way to earlier diagnosis. AB - Menkes disease (MNK) is a lethal, X-linked recessive disorder of copper metabolism dominated by neurodegenerative symptoms and connective tissue disturbances. The incidence of MNK in Asia is not known. Most patients die by the age of 3 years if adequate treatment is not carried out. Early parenteral administration of copper can prevent the neurological disturbances and lead to a better outcome. In the present study, a survey on MNK in Japan was performed. There were in total 53 live-born Japanese patients with MNK collected from 1990 to 2003, including two females. The incidence of live-born MNK patients between 1992 and 2002 was 2.8 per million live births (95% confidence interval (CI): 1.8 to 3.7), 4.9 per million male live births (95% CI: 3.2 to 6.6). One-third of the patients were born before 37 weeks or weighing less than 2500 g. Seventeen per cent were born both before 37 gestational weeks and weighing less than 2500 g. These proportions were higher than those in Japanese live-birth babies according to a nationwide estimate. The hair on these Japanese patients appeared not only as white or grey but also brown and blond. We also found that many signs had been noted before the patient was brought to a hospital with typical symptoms. These signs may be a clue to early diagnosis of MNK. PMID- 15902551 TI - Congenital cataract, muscular hypotonia, developmental delay and sensorineural hearing loss associated with a defect in copper metabolism. AB - Deficiencies of different proteins involved in copper metabolism have been reported to cause human diseases. Well-known syndromes, for example, are Menkes and Wilson diseases. Here we report a patient presenting with congenital cataract, severe muscular hypotonia, developmental delay, sensorineural hearing loss and cytochrome-c oxidase deficiency with repeatedly low copper and ceruloplasmin levels. These findings were suggestive of a copper metabolism disorder. In support of this, the patient's fibroblasts showed an increased copper uptake with normal retention. Detailed follow-up examinations were performed. Immunoblotting for several proteins including ATP7A (MNK or Menkes protein), ATP7B (Wilson protein) and SOD1 showed normal results, implying a copper metabolism defect other than Wilson or Menkes disease. Sequence analysis of ATOX1 and genes coding for proteins that are known to play a role in the mitochondrial copper metabolism (COI-III, SCO1, SCO2, COX11, COX17, COX19) revealed no mutations. Additional disease genes that have been associated with cytochrome-c oxidase deficiency were negative for mutations as well. As beneficial effects of copper histidinate supplementation have been reported in selected disorders of copper metabolism presenting with low serum copper and ceruloplasmin levels, we initiated a copper histidinate supplementation. Remarkable improvement of clinical symptoms was observed, with complete restoration of cytochrome-c oxidase activity in skeletal muscle. PMID- 15902552 TI - Preparation of 5-amino-4-imidazole-N-succinocarboxamide ribotide, 5-amino-4 imidazole-N-succinocarboxamide riboside and succinyladenosine, compounds usable in diagnosis and research of adenylosuccinate lyase deficiency. AB - The enzyme adenylosuccinate lyase (ADSL) intervenes twice in the biosynthesis of adenine nucleotides. ADSL deficiency is an inherited metabolic disease characterized by various degrees of psychomotor retardation and accumulation of dephosphorylated enzyme substrates 5-amino-4-imidazole-N-succinocarboxamide riboside (SAICAr) and succinyladenosine (SAdo) in body fluids. Severity of symptoms seems to correlate with residual activity of mutant enzyme and with SAdo/SAICAr concentration ratio in cerebrospinal fluid. To better understand the pathogenetic mechanisms of the disease symptoms, studies of catalytic properties of mutant enzymes together with in vitro and in vivo experiments utilizing SAICAr and SAdo must be performed. Such studies require availability of both ADSL substrates, 5-amino-4-imidazole-N-succinocarboxamide ribotide (SAICAR) and succinyladenosine 5'-monophosphate (SAMP) and their dephosphorylated products in sufficient amounts and purity. Except for SAMP, none of these compounds is commercially available and they must therefore be synthesized. SAICAR was prepared by recombinant human ADSL-catalysed reaction of AICAR (5-aminoimidazole 4-carboxamide) with fumarate and isolated by thin-layer chromatography. SAICAr and SAdo were prepared by calf intestine alkaline phosphatase-catalysed dephosphorylation of SAICAR and SAMP and isolated on cation- and anion-exchange resin columns. The procedures described are easily scalable and provide high yields of sufficiently pure products for use in experiments related to studies of pathogenetic mechanisms in ADSL deficiency. PMID- 15902553 TI - Inhibition of energy metabolism by 2-methylacetoacetate and 2-methyl-3 hydroxybutyrate in cerebral cortex of developing rats. AB - Mitochondrial beta-ketothiolase and 2-methyl-3-hydroxybutyryl-CoA dehydrogenase (MHBD) deficiencies are inherited neurometabolic disorders affecting isoleucine catabolism. Biochemically, beta-ketothiolase deficiency is characterized by intermittent ketoacidosis and urinary excretion of 2-methyl-acetoacetate (MAA), 2 methyl-3-hydroxybutyrate (MHB) and tiglylglycine (TG), whereas in MHBD deficiency only MHB and tiglylglycine accumulate. Lactic acid accumulation and excretion are also observed in these patients, being more pronounced in MHBD-deficient individuals, particularly during acute episodes of decompensation. Patients affected by MHBD deficiency usually manifest severe mental retardation and convulsions, whereas beta-ketothiolase-deficient patients present encephalopathic crises characterized by metabolic acidosis, vomiting and coma. Considering that the pathophysiological mechanisms responsible for the neurological alterations of these disorders are unknown and that lactic acidosis suggests an impairment of energy production, the objective of the present work was to investigate the in vitro effect of MAA and MHB, at concentrations varying from 0.01 to 1.0 mmol/L, on several parameters of energy metabolism in cerebral cortex from young rats. We observed that MAA markedly inhibited CO2 production from glucose, acetate and citrate at concentrations as low as 0.01 mmol/L. In addition, the activities of the respiratory chain complex II and succinate dehydrogenase were mildly inhibited by MAA. MHB, at 0.01 mmol/L and higher concentrations, strongly inhibited CO2 production from all tested substrates, as well as the respiratory chain complex IV activity. The other activities of the respiratory chain were not affected by these metabolites. The data indicate a marked blockage in the Krebs cycle and a mild inhibition of the respiratory chain caused by MAA and MHB. Furthermore, MHB inhibited total and mitochondrial creatine kinase activities, which was prevented by the use of the nitric-oxide synthase inhibitor L-NAME and glutathione (GSH). These data indicate that the effect of MHB on creatine kinase was probably mediated by oxidation or other modification of essential thiol groups of the enzyme by nitric oxide and other by-products derived from this organic acid. In contrast, MAA did not affect creatine kinase activity. Taken together, these observations indicate that aerobic energy metabolism is inhibited by MAA and to a greater extent by MHB, a fact that may be related to lactic acidaemia occurring in patients affected by MHBD and beta-ketothiolase deficiencies. If the in vitro effects detected in the present study also occur in vivo, it is tempting to speculate that they may contribute, at least in part, to the neurological dysfunction found in these disorders. PMID- 15902554 TI - Management of methylmalonic acidaemia by combined liver-kidney transplantation. AB - Methylmalonic acidaemia (MMA) is a rare autosomal recessive inborn error of metabolism that typically presents in infancy with recurrent episodes of metabolic acidosis, developmental delay and failure to thrive. The disease course is complicated by the development of chronic tubulointerstitial nephritis progressing to end-stage renal disease in adolescence. We describe two adolescents with cobalamin-nonresponsive MMA (mut0) who developed polyuria, chronic tubulointerstitial nephritis, dystonia but normal synthetic liver function. Both patients received combined liver-kidney transplantation (CLKT), preceded by a single pretransplant haemodialysis for clearance of methylmalonic acid. Post CLKT there was 95-97% reduction in serum and urine methylmalonic acid, leading to significant liberalization of dietary protein intake and a consequent increase in body mass index, muscle strength and energy. In addition, renal function normalized and clinical neurological status stabilized. We propose that CLKT be considered as a therapeutic option early in the course of cobalamin nonresponsive MMA. Progressive tubulointerstitial nephritis with disabling polyuria is a confounder in patient management even in the absence of end-stage renal disease. Successful CLKT restores methylmalonyl-CoA mutase enzyme levels in the liver and kidney, improves clearance of methylmalonic acid with resultant dietary protein liberalization, and offers excellent graft and patient outcomes with improvement in quality of life. PMID- 15902555 TI - OPA3 mutation screening in patients with unexplained 3-methylglutaconic aciduria. AB - We have screened 13 patients with neurological abnormalities and 3 methylglutaconic aciduria (3MGA) for mutations in the OPA3 gene, which are known to be the cause of Costeff syndrome (optic atrophy, chorea and spasticity; type III 3MGA). We aimed to explore whether mutations in the OPA3 gene are present in patients with 3MGA but without classic Costeff syndrome. OPA3 mutations (IVS1 1G>C) were identified in 2 patients with the classic phenotype of type III 3MGA, but not in the other 11 patients with differing non-Costeff phenotypes associated with developmental delay and neurological signs and symptoms as described. We identified a previously described sequence variation in the OPA3 gene (c.231T>C) in 12/13 patients. The alteration was homozygous in 8/12 and heterozygous in 4/12. This alteration was also found in 60 of 98 normal control alleles. In a single patient, a novel sequence variation in the 5' UTR was identified, (c. 38A>G). Our studies suggest that the c.231T>C sequence variation is of no clinical significance, whereas the significance of the 5' UTR sequence variation remains open to speculation. Our study of the OPA3 gene in patients with 3MGA without Costeff syndrome suggests that mutations in OPA3 are not a common cause of 3MGA in the absence of signs of Costeff syndrome. PMID- 15902556 TI - Biochemical, clinical and molecular findings in LCHAD and general mitochondrial trifunctional protein deficiency. AB - General mitochondrial trifunctional protein (TFP) deficiency leads to a wide clinical spectrum of disease ranging from severe neonatal/infantile cardiomyopathy and early death to mild chronic progressive sensorimotor poly neuropathy with episodic rhabdomyolysis. Isolated long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency resulting from the common Glu510Gln mutation usually gives rise to a moderately severe phenotype with multiorgan involvement with high morbidity and mortality. However, isolated LCHAD deficiency can also be consistent with long-term survival in patients identified and treated from an early age. We present biochemical, clinical and mutation data in 9 patients spanning the full spectrum of disease. Fibroblast acylcarnitine profiling shows good correlation with clinical phenotype using the ratio C18(OH)/(C14(OH)+C12(OH)). This ratio shows a gradation of values, from high in four patients with severe neonatal disease (2.5+/-0.8), to low in two neuromyopathic patients (0.35, 0.2). Fibroblast fatty acid oxidation flux assays also show correlation with the patient phenotype, when expressed either as percentage residual activity with palmitate or as a ratio of percentage activity of myristate/oleate (M/O ratio). Fibroblasts from four patients with severe neonatal disease gave an M/O ratio of 4.0+/-0.6 compared to 1.97 and 1.62 in two neuromyopathic patients. Specific enzyme assay of LCHAD and long-chain 3 ketothiolase activity in patient cells shows lack of correlation with phenotype. These results show that measurements in intact cells, which allow all determinative and modifying cellular factors to be present, better reflect patient phenotype. Mutation analysis reveals a number of alpha- and beta-subunit mutations. Peripheral sensorimotor polyneuropathy, often as the initial major presenting feature but usually later accompanied by episodic rhabdomyolysis, is a manifestation of mild TFP protein deficiency. The mild clinical presentation and relative difficulty in diagnosis suggest that this form of TFP is probably underdiagnosed. PMID- 15902557 TI - Normal acylcarnitine levels during confirmation of abnormal newborn screening in long-chain fatty acid oxidation defects. AB - We report two infants identified by tandem mass spectrometry (MS/MS) of neonatal blood spot acylcarnitines and confirmed by molecular genetic analysis to have long-chain fatty acid oxidation defects. In both cases, acylcarnitine concentrations in confirmatory plasma samples were normal. None the less, molecular testing identified trifunctional protein (TFP) deficiency (McKusick 600890) and very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency (McKusick 201475). PMID- 15902558 TI - Newborns with C8-acylcarnitine level over the 90th centile have an increased frequency of the common MCAD 985A>G mutation. AB - Medium chain acyl-coenzyme A dehydrogenase (MCAD) deficiency is the most commonly inherited defect of fatty acid oxidation. This autosomal recessive disorder is characterized by the tendency to become profoundly hypoglycaemic under fasting stress conditions, leading to lethargy, coma, brain injury and/or death. The most common mutation resulting in MCAD deficiency ascertained through individuals of northern European descent presenting with clinical symptoms is a single base-pair change (985A>G) that accounts for up to 90% of these abnormal alleles. In the general Nova Scotia population, which comprises largely individuals of northern European descent, this mutation is present at a frequency of 1 in 68. A recently implemented newborn screening programme for MCAD deficiency uses tandem mass spectrometry (MS/MS) to analyse blood spots from newborns for C8-acylcarnitine. After reviewing initial data from this newborn screening programme, we hypothesized that there was an unexpectedly high frequency of individuals with an MCAD 985A>G mutation and C8-acylcarnitine levels at the upper end of the normal range. A sample representing the upper 90th centile was screened for the presence of the 985A>Gmutation and 61 heterozygotes were identified, establishing a high frequency (1/10) of this 985A>G mutation in this selected population. We have demonstrated a relationship between heterozygosity for 985A>G and C8 acylcarnitine levels. These results contribute to the interpretation of C8 acylcarnitine levels and the establishment of a more clinically relevant screening cut-off point. PMID- 15902559 TI - The 625G>A SCAD gene variant is common but not associated with increased C4 carnitine in newborn blood spots. AB - The 625G>A variant of the short-chain acyl-CoA dehydrogenase (SCAD) gene is considered to confer susceptibility for developing 'clinical SCAD deficiency' and appears to be common in the general population. To determine the frequency of the 625G>A variant in The Netherlands, we analysed 1036 screening cards of 5- to 8 day-old newborns and found 5.5% homozygous and 31.3% heterozygous for the 625G>A variant. An increased blood/plasma C4-carnitine concentration is considered to be one of the biochemical characteristics of SCAD deficiency. To explore the correlation of C4-carnitine levels with the 625G>A variant, we determined the C4 carnitine concentration, as well as the ratio of C4- to free carnitine, in blood spots from newborns, who were detected as homozygous, heterozygous or noncarriers for the gene variant. No significant differences were found between these groups. Our study demonstrates a high frequency of the 625G>A SCAD gene variant in the Dutch population, but no correlation to significantly increased C4-carnitine levels in blood spots taken between the 5th and 8th days of life. This latter observation might be the result of the relatively late timing of neonatal screening in our country, implying that fatty acid oxidation disorders may be missed at that stage. If the 625G>A variant is associated with clinical SCAD deficiency, the high frequency of the variant suggests a possible involvement of SCAD deficiency in the pathogenesis of common disorders, probably in relation to other genetic and/or environmental factors. However, homozygosity for the 625G>A variant might be only a biochemical phenomenon, representing a 'nondisease'. PMID- 15902560 TI - Impaired myocardial perfusion reserve but preserved peripheral endothelial function in patients with Fabry disease. AB - Fabry disease (McKusick 301500) is an X-linked lysosomal storage disorder due to deficient alpha-galactosidase A activity, which leads to accumulation of glycosphingolipids, especially in vascular smooth-muscle and endothelial cells. The effect of this accumulation on peripheral and cardiac vascular function is poorly known. We studied 15 Fabry patients (mean age 35 years and mean BMI 24.8 kg/m2) and 30 age- and BMI-matched healthy controls to examine whether myocardial perfusion reserve and peripheral artery endothelial function are altered. Myocardial perfusion was measured at rest and during dipyridamole-induced hyperaemia by positron emission tomography and H2(15)O. Myocardial blood flow reserve was calculated as the ratio between the dipyridamole-induced maximal blood flow and resting blood flow. Peripheral artery endothelial function was assessed by measuring the brachial artery flow-mediated dilatation using ultrasound at rest and during reactive hyperaemia. The myocardial perfusion reserve was significantly lower in Fabry patients than in controls (3.3+/-1.2 vs 4.4+/-1.6, p=0.02), while the brachial artery flow-mediated dilatation was similar (5.9%+/-3.9% vs 4.5%+/-3.6%, p=0.27). Thus, inFabry disease, myocardial perfusion reserve is reduced while the peripheral artery endothelial function is preserved. PMID- 15902561 TI - Enzyme replacement therapy in Japanese Fabry disease patients: the results of a phase 2 bridging study. AB - Fabry Disease (alpha-galactosidase A deficiency) is an X-linked hereditary disorder leading to the pathological accumulation of globotriaosylceramide (GL-3) in lysosomes, particularly in the vascular endothelium of the kidney, heart and brain. We report the results of an open-label phase 2 study that was undertaken to evaluate whether ethnic differences exist that would affect agalsidase beta (Fabrazyme) treatment of Fabry patients in the Japanese population, relative to safety and efficacy. The study design mirrored the design of the completed phase 3 clinical trial that led to approval of the product agalsidase beta. The 13 Japanese, male Fabry patients enrolled in the study received the enzyme replacement therapy over a period of 20 weeks as biweekly infusions. All selected efficacy end points showed improvements that were comparable with findings from the phase 3 study. These improvements included reductions of GL-3 accumulation in both kidney and skin capillary endothelial cells to (near) normal levels (92% of patients). Kidney and plasma GL-3 levels decreased by 51.9% and 100%, respectively, by ELISA. Renal function remained normal. Fabry-associated pain, and quality of life, showed improvement over baseline in multiple categories. Related adverse events were mild or moderate in intensity and mostly infusion associated (fever and rigors). As expected, IgG antibody formation was observed in 85% of the patients, but had no effect on treatment response. These results suggest that treatment with agalsidase beta is safe and effective in Japanese patients with Fabry disease. With regard to safety and efficacy, no differences were observed as compared to the caucasian population. PMID- 15902562 TI - Evaluation of three biochemical markers in the monitoring of Gaucher disease. AB - Several markers have been developed for the biochemical monitoring of Gaucher disease. Three of the most commonly used markers are acid phosphatase, angiotensin-converting enzyme (ACE) and chitotriosidase. The rationale for using all three concurrently is not clear. A retrospective study was therefore carried out on data collected from 28 paediatric patients treated with enzyme replacement therapy. All three markers fell with time. However, chitotriosidase showed the steepest time trend, the largest trend by case interaction, and the lowest residual variance, making it the most reliable of the three. ACE correlated strongly with chitotriosidase, but acid phosphatase did not correlate well with either and also had the largest residual variance, indicating that it was too 'noisy' to be informative. The absence of a 'gold standard' for assessing Gaucher disease complicates the interpretation of these findings, but they suggest that acid phosphatase be dropped from routine clinical practice, and that chitriosidase be used in preference to ACE. PMID- 15902563 TI - Orthotopic liver transplantation from a living-related donor in an infant with a peroxisome biogenesis defect of the infantile Refsum disease type. AB - Peroxisomal biogenesis defects include a number of severe neurodevelopmental disorders, among which infantile Refsum disease (IRD) occupies the mildest end of the spectrum. Although high docosahexaenoic acid (DHA) and low phytanic acid diets can correct some of the biochemical defects, they have not consistently altered the progressive course of the disease. We carried out orthotopic liver transplantation (OLT) in a mildly symptomatic 6-month-old infant who was a sibling of a severely neurologically impaired older sister. After transplantation the clinical course of this young child appeared much improved by comparison to her older sister. She walked alone at 4 years, had acceptable social interaction and had a noticeable recovery of audition. After transplantation her biochemical parameters were significantly improved: phytanic acid and very long-chain fatty acid (VLCFA) serum concentrations decreased. Abnormal bile acids disappeared from plasma. Although the OLT did not result in a cure of the disorder, the clinical and biochemical results suggest that OLT should be considered in mildly symptomatic patients. PMID- 15902564 TI - Early diagnosis of mucopolysaccharidosis III A with a nonsense mutation and two de novo missense mutations in SGSH gene. AB - An early presentation of heparan N-sulphatase (SGSH) deficiency (mucopolysaccharidosis IIIA, MPS IIIA) with a prominent and isolated hepato splenomegaly is described. Molecular analysis detected a nonsense mutation (Y40X) and two de novo missense mutations (E300V; Q307P). PMID- 15902565 TI - Clinical response to sildenafil in pulmonary hypertension associated with Gaucher disease. AB - There are few reports of pulmonary hypertension in Gaucher disease. We report a patient who showed significant clinical improvement after treatment with sildenafil. PMID- 15902566 TI - Absolute configuration of N-acetylaspartate in urine from patients with Canavan disease. AB - High-resolution 1H and 13C NMR of N-acetylaspartic acid as its 2-(S)-butyl diester allows the two enantiomers to be unambiguously identified. Analysis of urine samples from five patients with Canavan disease (N-aspartoacylase deficiency) showed that more than 95% of the excreted N-acetylaspartate had the S configuration. PMID- 15902569 TI - [Botulinum toxin in neurogenic incontinence: decompression by intramuscular injection]. PMID- 15902570 TI - [Intersexuality syndrome: the undesired third sex]. PMID- 15902571 TI - [Urinary and fecal incontinence: incidence, symptoms and quality of life in the elderly]. PMID- 15902572 TI - [New targets and drugs for treatment of advanced renal cell carcinoma]. AB - Conventional immunotherapeutic approaches have failed to achieve fundamental benefits for clinical outcome of patients with advanced metastatic renal cell carcinoma (MRCC). New, encouraging substances have gained broad access to the field of oncology and have already shown most promising preliminary results in patients with MRCC. Of mayor interest are antibodies and (receptor) tyrosine kinase inhibitors that are targeted against growth-factor receptors (of vascular endothelial and/or tumor cells). Single or multi-drug regimens are under way and are soon to be marketed. PMID- 15902573 TI - [Long-term outcome of radical prostatectomy in an observation period of at least 10 years]. AB - PURPOSE: To evaluate the long-term outcome of radical prostatectomy in unselected patients with prostate cancer of relatively poor prognosis. MATERIALS AND METHODS: Between 1977 and 1989, 293 patients underwent radical retropubic prostatectomy. We analyzed 224 patients with a minimum follow-up of 10 years (mean 14 years, range 10.5 to 24 years). RESULTS: The pathological stage distribution of these 224 patients was pT1 in 14.3 %, pT2 43.6 %, pT3 39.1 % and pT4 3 %. Of these patients, 50 (22.3 %) had positive lymph nodes and 45 (20.1 %) positive margins. At 10 years after surgery, the actuarial survival rate for all patients was 67.1 % and the cancer-specific survival rate 85 %. Once progression of the disease developed, the median actuarial time to death was 5.5 years. Since PSA measurements were not introduced in the follow-up after radical prostatectomy until 1986, PSA data were available in only 84 (37.5 %) patients. In a survival analysis, preoperative PSA (< 10 ng/ml vs. > or = 10 ng/ml) was a significant predictor of the probability of positive margins (22 % vs. 78 %) and cancer specific survival (87 % vs. 69 %) after 10 years. CONCLUSIONS: Long-term survival can be expected even in patients with locally advanced prostate cancer treated with radical prostatectomy. PMID- 15902574 TI - [Are there indications for organ-preserving tumor resection in urothelial cancers of the upper urinary tract?]. AB - PURPOSE: Identification of prognostic factors for tumor recurrence in nephronsparing surgery and tumor progression after organ-preserving surgery. MATERIAL AND METHODS: From June 1989 to February 2003, 43 patients (47 nephron units) underwent organ-preserving surgery or the upper urinary tract because of urothelial carcinoma, with 26 (60.5 %) having an elective indication (healthy contralateral kidney). RESULTS: The mean follow-up was 39.2 months with a range of 10.6 to 168.3 months. Ipsilateral recurrences occurred in 13 (27.6 %) of reno ureteral units. Recurrences were significantly more common for tumors located in the renal pelvis or opposite the ureteral origin (p = 0.018). Tumor progression occurred in 8 of 43 patients (18.6 %) and significantly correlated with the T- and G-stage of the primary tumor (p = 0.006 and p = 0.002). Of the 47 conservatively treated reno-ureteral units, 38 (80.8 %) could be preserved. CONCLUSION: Organ-preserving resection of the urothelial carcinoma of the upper urinary tract is an alternative in some patients with well-differentiated superficial tumors. A follow-up for life and a good patient compliance are necessary. Tumors of the renal pelvis have a significantly higher recurrence rate than ureteral tumors. Poorly differentiated tumors should undergo organ preserving surgery only if the goal is palliation and in patients with solitary kidney after intensive consultation about the high risk of invasive recurrences and the development of metastases. PMID- 15902575 TI - [Ki-67 antisense therapy in murine renal cell carcinoma models]. AB - PURPOSE: The Ki-67 antigen is only expressed in proliferating cells. Previously, it was shown that Ki-67 derived antisense oligonucleotides (asONs) specifically inhibit the proliferation of tumor cells and tumour growth in vitro and in subcutaneous bladder and prostate tumor models. We intended to evaluate the effects of this therapeutic concept in two renal cell carcinoma (RCC) models. MATERIAL AND METHODS: Human RCC cells (SK-RC 35) were initially transfected with FITC-labeled ONs and diffferent cationic lipids to analyze the transfection efficacy by flow cytometry (FACS). The potency of 14 different ONs sequences was compared by quantitative RT-PCR in vitro. For in vivo testing, ONs were administered to immunocompetent Balb/c mice bearing orthotopic RENCA tumors as well as to SCID mice bearing subcutaneous RCC SK-RC 35 xenografts. Tumor sizes and final tumor weights were documented. Additionally, several immunohistochemical staining procedures were performed. RESULTS: FACS analysis showed highly effective transfection conditions in vitro. Systemic administration of asONs significantly decreased the tumour growth in the RENCA model (p < 0.05) and in the SCID mouse model (p = 0.009). Immunohistochemical staining of tumor specimens revealed a marked down-regulation of target protein and a slight increase in apoptotic cells after antisense treatment while the microvessel count was not significantly altered. CONCLUSION: These results demonstrate that the Ki 67 antigen represents a suitable antiproliferative target and that asONs directed against this target are potent drugs that induce a significant inhibition of renal tumor growth in different mouse models. PMID- 15902576 TI - [Prostate biopsy -- practical examination of the adequacy of Chen's virtual strategy]. AB - INTRODUCTION: Prostate sextant biopsy is considered the gold standard of invasive prostate cancer diagnostics. However, numerous studies have shown that greater detection rates are achieved by increasing the number of biopsies. Chen et al. found ten "ideal" biopsy sites using a computer model. This study examines the clinical value of this variation of number and localisation of biopsies. MATERIAL AND METHOD: Ultrasound-guided ten-core prostate biopsies are performed in our department. Following the model of Chen, three prostate biopsies instead of on are taken from the middle plane. The two additional specimens are obtained in the paraurethral and ventral regions. RESULTS: Between July 2003 and February 2004, 191 patients scheduled for a 10-core prostate biopsy were included in the study. Assessable results were obtained in 176 cases. A carcinoma was detected in 61/176 (34.7 %) of the patients. The carcinoma was found only in the standard sextant localisations (27.8 %) in 17/61 patients and additionally in the paraurethral or ventral region in 35/61 (57.3 %). The carcinoma was located only paraurethrally in 1/62 (1.6 %) and only ventrally in 6/62 (9.7 %). Altogether, the prostate carcinoma was detected only by the additional biopsies in 6 patients (9.8 %). A clinically relevant disease was seen in 5/6 (83.3 %). A carcinoma was detected most frequently in the following biopsies: base right (17.1 %), central lat. right (17.5 %) and apex right (16.5 %) and most rarely paraurethral right (12 %) and ventral right (10.2 %). Statistically significant differences were found only between these groups. CONCLUSION: The modification and increase of biopsies from six to ten according to the model suggested by Chen et al. detected 9.8 % more carcinomas in our patients. PMID- 15902577 TI - [Laparoscopic colpopromontofixation]. PMID- 15902578 TI - [Abdominal colpopromontofixation]. PMID- 15902579 TI - Pharmacophilia and pharmacophobia: determinants of patients' attitudes towards antipsychotic medication. AB - OBJECTIVE: To identify factors that influence attitudes towards psychopharmacological treatment in patients suffering from schizophrenia and schizoaffective psychoses. METHODS: Ninety-two participants in an outpatient psychoeducational program, classed as "pharmacophobic" or "pharmacophilic" according to the Drug Attitude Inventory scale, were compared with regard to sociodemographic variables, clinical characteristics, subjective deficit syndrome, illness concepts, knowledge, locus of control, and quality of life. RESULTS: The 59 pharmacophilic and the 33 pharmacophobic patients did not differ significantly with regard to most sociodemographic variables, symptoms, or classic personality traits such as locus of control, self-concept, and quality of life. The only differences concerned hospitalization history ( P < 0.05) and statements on the actual, subjective experience of desired and undesired effects of medication ( P < 0.01). CONCLUSIONS: The impact of subjective experiences with drug treatment on attitudes towards medication and compliance needs to be a main focus of interventions targeting attitudes towards pharmacological treatment. PMID- 15902580 TI - Relationship between mirtazapine dose, plasma concentration, response, and side effects in clinical practice. AB - OBJECTIVE: The aim of this study was to evaluate the benefit of mirtazapine plasma concentration monitoring in a typical clinical setting. METHODS: The relationship between mirtazapine plasma concentration, dose, response, and side effects was studied in 65 inpatients presenting with a depressive episode according to ICD-10. Plasma concentrations, the 17-item Hamilton Depression Rating (HAMD), and the UKU side effect rating were performed weekly. A subgroup of 45 patients was evaluated for a concentration-response relationship. RESULTS: We found a low positive correlation between plasma concentration and dose. A low negative correlation between plasma concentration and increased duration of sleep was noted in the first week of mirtazapine treatment, but not during the entire observation time. Responders to mirtazapine treatment presented with higher plasma concentrations than non-responders, revealing a threshold concentration of 30 ng/mL. CONCLUSION: The mirtazapine dose is a weak predictor of mirtazapine plasma concentrations. Plasma concentration measurements may therefore be useful to adjust mirtazapine doses in non-responders with plasma concentrations below 30 ng/mL. Sedative effects appear temporary and require no plasma concentration control when standard doses are administered. PMID- 15902581 TI - Paroxetine serum concentrations in depressed patients and response to treatment. AB - INTRODUCTION: There is no established relationship between the serum concentration of selective serotonin reuptake inhibitors (SSRIs) and clinical response in depressed patients. METHODS: We analyzed paroxetine concentrations in serum of 46 depressed patients during treatment with a fixed dosage of 40 mg paroxetine. RESULTS: After 5 weeks 29 patients responded to treatment, while 17 did not. Analysis of variance with repeated measures (ANOVA-rm) revealed a significant effect of "response" with responders having lower serum concentrations throughout the treatment period, when compared to non-responders. After 2, 3, and 4 weeks of treatment, we could define an upper threshold of paroxetine serum concentrations (week 1 : 22.7 ng/mL; week 2 : 43 ng/mL; week 3 : 53.4 ng/mL; week 4 : 39.1 ng/mL) above which response to treatment was unlikely. CONCLUSION: We conclude that -- in contrast to other pharmacological approaches - high rather than low drug serum concentrations may be associated with non response in paroxetine treatment of depressed patients. PMID- 15902582 TI - Reasons for switching between antipsychotics in daily clinical practice. AB - BACKGROUND: Previous research that atypical antipsychotics were switched less often compared to typical antipsychotics, suggesting overall better treatment satisfaction. The objective of this study was to investigate the reasons for switching antipsychotics after initiating oral treatment with either typical or atypical antipsychotics in a clinical setting. PATIENTS AND METHODS: A total of 123 patients that switched antipsychotic therapy were recruited from 17 psychiatric hospitals, of which 46 % switched because of lack of effect and 45 % because of adverse effects. RESULTS: No significant differences were found between users of atypical versus typical antipsychotics in reasons for switching, both for overall adverse events, and lack of effect. In users of atypical antipsychotics extrapyramidal effects were reported less often as reason for switching (adjusted OR = 0.18 (95 % CI = (0.07 - 0.51)). Patients on atypical antipsychotics switched more often because of weight gain (adjusted OR = 12.8 (95 % CI = (1.50 - 109)). CONCLUSION: In conclusion, when switching occurred, no difference was found in the frequency of general tolerability or reported lack of effectiveness. However, the type of adverse event as a reason for switching differed between atypical and typical antipsychotics. PMID- 15902583 TI - Dyspepsia in chronic psychiatric patients. AB - INTRODUCTION: We report on dyspeptic complaints among patients hospitalized in the long-stay ward of a general psychiatric hospital. METHODS: A representative sample of the patients was interviewed using a structured questionnaire. RESULTS: Eighty percent of the patients reported one or more symptoms of dyspepsia, and 68 % reported symptoms of reflux-like dyspepsia. CONCLUSION: Significant positive associations were found for dyspepsia complaints and clozapine (OR = 3.4), laxatives (OR = 4.4), and heavy smoking (OR = 2.3). PMID- 15902584 TI - Epidemiology of first- and second-generation antipsychotic agents in Lombardy, Italy. AB - INTRODUCTION: This study investigated the age and gender distribution of antipsychotic prescribing in Lombardy, a region of nine million inhabitants in northern Italy. METHODS: From the Regional Administrative Database of Lombardy, all ambulatory prescriptions of antipsychotics dispensed during 2001 were extracted and prevalence data were calculated by dividing users by the total number of male and female residents in each age group. RESULTS: During the study period 86,187 subjects were dispensed antipsychotic agents, yielding a prevalence of use of 0.87 (95 % CI: 0.86, 0.88) per 100 males and 1.01 (95 % CI: 1.00, 1.02) per 100 females. The prevalence of use progressively rose with age in both sexes, with the highest rates in old and very old subjects. The prevalence of use of first-generation antipsychotics progressively increased with age and dramatically increased in old and very old subjects; in contrast, the prevalence of use of second-generation antipsychotics remained substantially stable or slightly decreased up to 65 years of age and increased thereafter. CONCLUSION: Antipsychotic agents are prescribed widely in the general population, and very high rates were observed in those aged 80 years or more. PMID- 15902585 TI - Correlation of subjective well-being in schizophrenic patients with gait parameters, expert-rated motor disturbances, and psychopathological status. AB - INTRODUCTION: Subjective well-being of schizophrenic patients can be impaired by symptoms of the disease and by adverse effects of antipsychotic medication. We assessed the correlations of subjective well-being with objectively measured gait parameters, expert-rated motor disturbances, and psychopathological status in 25 conventionally treated, 25 atypically treated, and 16 drug-naive patients. METHODS: Main variables were the SWN scores (Subjective Well-being under Neuroleptic Treatment Scale), the ESRS scores (Extrapyramidal Symptoms Rating Scale), and the PANSS scores (Positive and Negative Syndrome Scale). Gait parameters were determined by using an ultrasonic system for gait analysis. RESULTS: In conventionally treated patients, the SWN total score significantly correlated with stride length ( R(2) = 0.39; P < 0.01), whereas in atypically treated and drug-naive patients, it significantly correlated with the PANSS score (atypically treated: R(2) = 0.25, P < 0.05; drug-naive: R(2) = 0.64, P < 0.01), mainly due to the correlations with the "negative symptoms" and the "general psychopathology" sub-scores. Correlations with stride length were significant not only in the "physical functioning" sub-score of the SWN but also in all other sub scores. Correlations of the SWN scores with ESRS scores were weak. CONCLUSION: Under conventional antipsychotic treatment, subjective well-being particularly depends on major side effects, whereas in atypically treated and drug-naive schizophrenic patients, it is mainly influenced by psychopathological status. Motor adverse effects of conventional antipsychotic treatment cannot be considered as isolated physical side effects but have severe implications for other aspects of the patients' well-being. PMID- 15902586 TI - The intensity dependence of auditory evoked ERP components predicts responsiveness to reboxetine treatment in major depression. AB - INTRODUCTION: The intensity (loudness)-dependent amplitude change (IDAP) of auditory evoked event-related potential (ERP) components has been suggested as an indicator of central serotonergic neurotransmission. In patients with major depression, associations of high IDAP with favorable SSRI treatment outcome have been reported. This is the first study to assess the predictive value of the IDAP in SNRI treatment. METHODS: We evaluated the pre-treatment intensity-dependent change of auditory evoked P1, N1, P2, and P1/N1 and N1/P2 peak-to-peak amplitudes in 14 inpatients with major depressive episode (DSM IV) in the course of 24 days of treatment with the SNRI reboxetine (6-12 mg/d). RESULTS: Our data revealed a highly significant correlation between lower intensity-dependent N1 amplitude slopes prior to reboxetine treatment and stronger decrease of HDRS total score at Fz ( r = 0.86, P < 0.001), Fcz ( r = 0.91, P < 0.001), and Cz ( r = 0.93, P < 0.001). CONCLUSION: This result corroborates the hypothesis of the IDAP as a differential indicator of serotonergic versus noradrenergic antidepressant psychopharmacotherapy. PMID- 15902587 TI - Successful treatment of sexual disinhibition in dementia with carbamazepine -- a case report. AB - Sexual disinhibition is a disturbing behavioral symptom in dementia. At present, there are no treatment guidelines available. Here we present the successful use of carbamazepine in a 78-year old AD patient with hypersexual behavior. The efficacy of carbamazepine in this case is in parallels to its effects on aggression and agitation in dementia and supports the important role of anticonvulsants in the management of behavioral disturbances in demented patients. PMID- 15902588 TI - Severe diurnal somnolence induced by fluvoxamine-lithium combination. PMID- 15902589 TI - [Corticosteroid therapy in ARDS--significant mortality]. PMID- 15902590 TI - [Common cold--Italians suffer the most]. PMID- 15902591 TI - [COPD--decided thickening of the small bronchial pathways]. PMID- 15902592 TI - [Prognosis for patients with COPD]. PMID- 15902593 TI - [Lung matters--help for the patient with stomach problems?]. PMID- 15902594 TI - [It is still coming: molecular-targeted therapy of lung cancer]. PMID- 15902595 TI - [Diaphragmatic weakness and its impact on respiratory function in primary myopathies]. AB - BACKGROUND: Diaphragmatic weakness (DW) is a potential manifestation of primary myopathies. Prevalence and impact on respiratory function have not been defined. METHODS: Respiratory function (inspiratory vital capacity, IVC; maximal inspiratory muscle pressure, PImax; respiratory muscle effort, P (0.1)/PImax) and polysomnography/nocturnal capnometry (PtcCO(2)) was analysed in 49 patients with primary myopathies. DW was defined as > 25 % drop of IVC upon shift from upright to supine position. RESULTS: 19/54 (35 %) of patients, mostly AMD (68 %) and DMD (42 %) had DW. Restrictive ventilatory defect was moderate (IVC 37.2 +/- 26.2 %) in patients with and without DW (IVC 46 +/- 26 %, p = n. s.). DW caused a -33 %, respective - 25 %, supine drop of IVC and PImax, resulting in severe restriction in supine position (IVC (supine) 24.9 +/- 19.1 %, PImax (supine) 2.0 +/- 1.0 kPa, P (0.1)/PImax (supine) 19.9 +/- 12,8 %). All patients with DW and 75 % without had sleep-disordered breathing. This was significantly more severe in DW: 90 % (vs 17 % without DW) had continuous nocturnal hypoventilation (PtcCO(2) 62 +/- 2 mm Hg), 70 % (vs 18 % without DW) had combined nocturnal and diurnal hypercapnic respiratory failure (PaCO(2) 54,7 +/- 11.8 mm Hg). DW was an independent risk factor for sleep disordered breathing, for nocturnal and diurnal respiratory failure (r = 0.95, p < 0,05). Predictive thresholds thereof were accurate and identical to previously determined only for supine function data. CONCLUSION: DW is common in primary myopathies and predictive of nocturnal and diurnal respiratory failure. Supine respiratory function tests are necessary for clinical diagnosis and respiratory risk stratification. PMID- 15902596 TI - [Inhalation therapy by dose-inhalers: analysis of patients performance and possibilities for improvement]. AB - Inhalation therapy in chronic obstructive airways disease requires an efficient inhalation technique. This study analyses step by step the mistakes made in the usage of different MDIs, relates these to patient information prior to the testing and examines several teaching procedures for improvement of knowledge and performance of the inhalation technique. 125 patients suffering from COPD were assigned to six different groups according to their background knowledge in the inhalation technique. The performance was assessed in standardized single steps and as overall performance. Furthermore the efficacy of an interactive pc-based training program was evaluated. The worst performance was seen in patients who only used the suppliers medication leaflet as a guide. Patients trained in outpatient clinics as well as patients trained in small groups during an inpatient stay showed a better performance. A high improvement rate was seen in prior MDI naive patients after they had undergone the interactive pc-based training program. Most problems were detected in the application step "exhalation before inhalation" and in the actuation-inhalation step. Besides the classical and the pc-based training the use of MDI phantoms showed very good results. The practical conclusion of this study is that the ability of patients to use inhalation pharmacotherapy efficiently needs improvement. Training programs of different intensity lead to a different outcome in performance and knowledge depending on prior knowledge. Inhalation pharmacotherapy without adequate training is insufficient. PMID- 15902597 TI - [Non-small-cell lung cancer third-line therapy with gefitinib]. AB - The EGFR-inhibition via tyrosine-kinase-inhibitor gefitinib (Iressa) constitutes a new way to treat non-small-cell lung cancer. Recent research results enable us to better understand the basics of molecular targeted therapy (MTT). These results are helpful to re-interpret the clinical results obtained so far for gefitinib and to consider for the first time in a predictive manner factors for the selection of patients suitable for therapy. Three case reports are presented in this paper which illustrate that -- with view to the results from translational research -- the use of gefitinib offers an efficient new therapeutic modality for the treatment of chemotherapy-resistant non-small-cell lung cancer. PMID- 15902598 TI - [Possibilities and perspectives of home-based exercise training in patients with chronic obstructive pulmonary diseases]. AB - In Germany there is a long tradition of rehabilitation and exercise training programs to improve the clinical state and physical function of patients with chronic obstructive pulmonary diseases. This is particularly true regarding inpatient rehabilitation. Based on experiences gained in other countries, outpatient and home-based programs are currently getting more attention. One of the key components of pulmonary rehabilitation is exercise training which can be expected to exert direct and indirect beneficial effects on physical performance, health-related quality of life and even the progress of the disease. Most of the available experience refers to patients with COPD -- besides those with bronchial asthma. The data available so far suggest that exercise training programs can reduce exacerbation rates and that this effect is achievable by minimal personal assistance. Thus the most important aim of future studies seems to be the development and evaluation of training programs which are efficient but also easy to implement and minimally expensive. Furthermore it needs to be studied whether such programs can be transferred to pulmonary disorders other than obstructive airway diseases. PMID- 15902599 TI - [Therapeutic options in malignant pleural mesothelioma]. AB - Malignant pleural mesothelioma may be treated with surgery, radiotherapy and chemotherapy. In most patients, the treatment remains palliative with symptom relief and a moderate survival gain. Only a minority of patients with early stage mesothelioma may be cured by a multimodal approach including radical surgery, chemotherapy, and radiotherapy. We discuss the role of surgery with either radical extrapleural pleuropneumonectomy or less invasive palliative pleurectomy and decortication, and the role of radiotherapy, in which the main problem is how to deliver sufficient doses to the pleural surface, sparing radiosensitive structures such as the lung, heart, liver, and kidneys. Chemotherapeutic options are discussed with 'older' mono- and combination regimens and the new promising combination cisplatinum/pemetrexed, now the 'standard regimen' for malignant pleural mesothelioma. Experimental approaches such as hyperthermia, interferons or interleukins, and 'small molecules' or antibodies inhibiting the EGFR oder VEGF receptor are under clinical evaluation. For the majority of our patients we recommend talcum pleurodesis either by medical thoracoscopy or VATS, followed by chemotherapy with platinum/pemetrexed. Radiotherapy may be applied in case of local tumour growth. The individual therapeutic decision will depend on tumour stage, concomitant diseases, performance status, and on the patient's preference. PMID- 15902600 TI - [Initiator of the open-air rest cure: Peter Dettweiler and the sanatorium at Falkenstein (Taunus) in Germany]. PMID- 15902601 TI - [Intensive care medicine -- an ethical challenge]. PMID- 15902602 TI - [Prof. Dr. med. Dr. h. c. Gunter Hempelmann on his 65th Birthday]. PMID- 15902603 TI - [Pain on injection with propofol]. AB - Injection-induced pain during induction of anaesthesia can result in patient's discomfort. This can prevent the use of propofol-anaesthesia in paediatric patients. Because of the high incidence of pain on injection with propofol numerous interventions have been tested to prevent this pain including the use of different drugs and physical measures as well as the combination of methods. The use of a single intervention is not as effective as the combination of different preventive measures. Thus the additional application of a venous tourniquet improves the pain reducing effect if drugs with peripheral mechanism of action are used for prevention of pain. Injection of lidocaine with a rubber tourniquet before the propofol injection is recommended as best effective method. In any case a propofol-MCT/LCT-Emulsion should be used for propofol-anaesthesia. Additionally a multimodal strategy that is adapted to the daily clinical practice seems to be convenient. That means, for general anaesthesia opioids or ketamine and for sedation a sub-anaesthetic dose of thiopental can be used for effective prevention of pain. If the prevention of nausea and emesis is intended, antiemetics that are appropriate for prevention of injection pain should be given. For paediatric anaesthesia the application of EMLA-cream seems to be suitable, because it alleviates the pain during venous cannulation at the same time. Although a painfree propofol injection is not possible with every prevention strategy there exist a wide range of effective interventions to prevent pain on injection with propofol. This article reviews methods that have been investigated and established to minimise the incidence of pain. PMID- 15902604 TI - [Mortality in German intensive care units: dying from or with a nosocomial infection?]. AB - OBJECTIVE: To describe mortality from intensive care unit acquired infections in German ICUs. METHODS: The hospital infection surveillance system (KISS = Krankenhaus-Infektions-Surveillance-System) started in Germany in January 1997 as a nationwide surveillance project for the voluntary registration of nosocomial infections. For ICUs reporting focuses on pneumonia and primary bloodstream infections. The data collected for each patient who acquires a nosocomial infection include outcome information. Data of a recent French study and the Statistical Yearbook were used to estimate the annual number of ICU acquired infections contributing to death in Germany. RESULTS: Data from 897 774 ICU patients were considered for the analysis (January 1997-June 2004). ICU mortality of patients with nosocomial pneumonia was 8.8 %, of patients with primary bloodstream infection was 10,9 %. Of the ICU patients with nosocomial pneumonia and primary bloodstream infections about 3554 die annually, in about 1131 cases the nosocomial infection was the cause of death. It may be estimated that the ICU acquired cases of pneumonia and primary bloodstream infection contributed to the death of about 2400 patients annually in Germany. CONCLUSION: Because of the substantial mortality of patients with ICU acquired infections preventive measures have a high priority. Further studies leading to improved infection control measures are very wellcome. PMID- 15902605 TI - [Hotline 2: heating capabilities of a coaxial fluid warming system at low and moderate flow rates]. AB - OBJECTIVE: The aim of this study was to compare the effects of two coaxial fluid warming systems with their heating capabilities. METHODS: The heating capabilities of two coaxial fluid warming systems and their capabilities to warm fluids at 20 +/- 0.5 degrees C (20 degrees C room temperature) was measured: 1) Hotline 1 Fluid Warmer, 2) Hotline(R) 1 Fluid Warmer. Final temperatures were measured at different infusion rates (10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 400, 800, 1000 ml/h and at maximal flow rates (Vmax)), at the distal end of the disposable tubing. Statistical analysis was performed using a computer based program (NCSS). Differences between the groups were analysed using the Two-Way ANOVA. Significance was defined at a p < 0.05. RESULTS: At flow rates between 10 1000 ml/h, infusion temperatures of > 36 degrees C were attained by both devices in a reliable manner. Compared to Hotline 1, Hotline 2 attained higher final temperatures of between 1.2 - 3.8 % (p < 0.01). Hotline 1 was measured to have higher Vmax (+ 3.6 %) compared to Hotline 2 (p < 0.01). However, the mean final temperature at Vmax of Hotline 2 was increased by 6.5 % (p < 0.01). The time needed to warm target temperature to 40 - 41 degrees C took between 9 to 12 minutes (flow rate 10 ml/h and Vmax respectively) when using Hotline 1 and 3 to 4 minutes when using Hotline 2. CONCLUSION: Compared to its predecessor, the Hotline 2 performance is a valuable improvement with respect to heating capacity and clinical handling. PMID- 15902606 TI - [The patient at increased cardiac risk -- are there new aspects?]. PMID- 15902607 TI - [Preoperative cardiovascular evaluation before non-cardial surgery]. PMID- 15902608 TI - [Preoperative management of patients with increased cardiac risk]. PMID- 15902609 TI - [Which monitoring, when and to which use?]. PMID- 15902610 TI - [Anaesthesia in patients with pacemaker or cardioverter]. PMID- 15902611 TI - [Options and limitations of pre-clinical diagnostics and treatment]. PMID- 15902612 TI - [End of life decisions in intensive care]. PMID- 15902617 TI - [Is measuring by portable heart rate monitors (sport watches) dependable in patients with an implanted pacemaker?]. AB - BACKGROUND AND OBJECTIVE: Regular physical exercise is advantageous for physical fitness and endurance. The intensity of exercising can be controlled by portable heart rate monitors. The aim of the study was to investigate if wearable monitors work properly in pacemaker patients. PATIENTS AND METHODS: In 30 patients with an implanted single or dual chamber pacemaker made by five different manufacturers their heart rate was determined simultaneously by surface ECG and Holter ECG, and was compared with the heart rates derived from portable heart rate monitors (Polar A1) at rest, during 6 minutes in-house walk, and while exercising at 25, 50, and 75 watts in a bicycle exercise test. The tests were done with pacing set at uni- as well as bipolar stimulation mode. RESULTS: At rest in two of 30 patients (6,6 %) paced in unipolar mode the portable heart rate monitors counted double. Under exercise conditions the portable monitors counted double in six of 26 patients (23 %) while being stimulated in unipolar mode. In bipolar stimulation mode, the portable heart rate monitors were working properly in all patients under all conditions. CONCLUSIONS: Patients with an implanted bipolar pacing system can control their physical exercising safely and accurately by means of portable heart rate monitors. Pacemaker patients who are planning regular physical exercising should be provided with an implanted bipolar pacemaker system. PMID- 15902618 TI - [Asymptomatic myocarditis after infection of the upper respiratory tract]. AB - HISTORY AND CLINICAL FINDINGS: A 20-year-old patient was hospitalized with persistent high fever after tonsillitis and swelling of the talocalcanean joint. INVESTIGATIONS: The ECG showed a partial right bundle branch block pattern and pathological T inversions on the left precordial leads. Cardiac Troponin I levels were slightly elevated and echocardiography revealed a dyscinetic area at the right ventricular apex. The anti-streptolysin titer was elevated. DIAGNOSIS: Post streptococcal rheumatic myocarditis. THERAPY AND FOLLOW-UP: Antibiotic therapy for 2 weeks. The patient showed subjective full recovery after 6 weeks. The anti streptolysin titer further increases. Nuclear spin tomography of the heart reveal postinflammatory alterations at the apex of the right ventricle. CONCLUSION: Rheumatoid fever is a rare diagnosis in developed countries. This case, however, illustrates that the true prevalence of rheumatoid carditis might be underestimated in the presence of only minimal heart-and joint-specific symptoms. PMID- 15902619 TI - [Rare cause of right heart failure: two cases]. AB - HISTORY AND ADMISSION FINDINGS: Two asylum seekers (patient A, 30 year old man from Mongolia; patient B, 18 year old woman from the Sudan) were referred to our outpatient clinic because of acute and chronic deterioration of their general condition and shortness of breath. Both patients presented with a clear clinical picture of systemic venous hypertension and moderate pulmonary congestion. Patient B had a paradoxical pulse compatible to cardiac tamponade. INVESTIGATIONS: In patient A, the chest radiogram revealed a markedly enlarged cardiac silhouette and an infiltrate in the upper left lobe of the lung. The echocardiogram confirmed a pericardial effusion causing a cardiac tamponade. A therapeutic and diagnostic pericardiocentesis was performed immediately. In patient B, the chest radiogram revealed a thickened and calcified pericardium and a left-sided pleural effusion. The pleural fluid revealed a lymphocyte predominant exudate. In both patients the tuberculin skin test was positive. DIAGNOSIS, TREATMENT AND COURSE: In both patients we initiated an antituberculous therapy (four-drug therapy with isoniazid, rifampin, pyrazinamide and ethambutol; in patient A in addition corticosteroids). In patient A the PCR for Mycobacterium tuberculosis complex from pericardial fluid was positive. In patient B we started the antibiotic treatment despite negative microbiological studies because of the high degree of suspicion in a person at high risk. Due to the impaired ventricular filling, patient B required additional pericardectomy. After completion of treatment, both patients have been doing well. CONCLUSION: Despite a clear reduction in the incidence of tuberculous pericarditis in Europe this multifaceted condition should still be of concern in patients at high risk as e. g. immigrants from areas with a high tuberculosis prevalence. Early diagnosis and adequate treatment are required for prevention of severe complications and disabling constrictive pericarditis. PMID- 15902620 TI - [Tyrosine kinase inhibitors in tumor therapy--part 1. Molecular and genetic fundamentals]. PMID- 15902621 TI - [Antiarrhythmic therapy--recent aspects of pharmacologic treatment]. AB - Over the last years the indication for antiarrhythmic therapy has changed due to the development of other therapeutic approaches. However, antiarrhythmics are important in the acute treatment as well as the prevention of recurrent rhythm disorders. In line with the antiarrhythmic agents of class IC and III also beta blockers, ACE inhibitors and AT (1) antagonists can be used primarily with a lower risk of severe cardiac side effects. Recent studies demonstrate that for patients with atrial fibrillation there was no benefit of rhythm control versus rate control. However, rhythm control with antiarrhythmics is beneficial in the treatment of highly symptomatic or hemodynamically compromised patients. Hybrid therapy and the "pill in the pocket"-strategy seem to be potent new therapeutic options. PMID- 15902622 TI - [Elective therapy of asymptomatic infrarenal aortic aneurysms. Comparison of mortality and morbidity between endovascular and open repair]. AB - Summary. In recent years, endovascular operations on infrarenal aortic aneurysm (EVAR) have not only been acknowledged as an alternative, but have become an indispensable "tool of the trade" for the vascular surgeon. As documented in many studies, perioperative morbidity is definitely much lower than in open surgery. In the long term, open surgery is also subject to complications such as rupture or aorto-enteral fistulae which necessitate secondary operations with their associated risks. There are some valid studies documenting a significantly lower mortality for EVAR than for open surgery. These datas were recently confirmed with high evidence level by the published results of UK-EVAR trial 1. The other randomized trials are currently ongoing. If the lower morbidity and lethality rate for EVAR is confirmed in these studies, the establishment of the indication for aortic aneurysm must be reconsidered with regard to the risk profile and the maximum diameter. The fact that long-term results are not yet available for the endovascular operation introduces a factor of uncertainty. Intensive follow-up observation is therefore an absolute prerequisite in patients who have undergone endovascular treatment. If the establishment of the indication is appropriate, with the new vascular prosthesis endoleaks are no longer an insoluble problem. The two methods should not be regarded as mutually competitive, but as complementary components of the treatment spectrum at every vascular center. PMID- 15902623 TI - [TSH degradation by stress?]. PMID- 15902625 TI - [Diagnosis of chronic mesenterial artery ischemia. Re the article from DMW 3/2005]. PMID- 15902626 TI - [MRI for troubleshooting detection of prostate cancer]. AB - Prostate cancer is the most common malignancy in males. Men aged 50 and older are recommended to undergo an annual digital rectal examination (DRE) and determination of prostate-specific antigen (PSA) in serum for early detection. However, prostate biopsies guided by transrectal ultrasound (TRUS) come up negative for cancer in many patients despite having PSA levels above 4 ng/ml. T2 weighted Magnetic Resonance Imaging (MRI) is able to represent the prostate including the surrounding anatomy and depict suspicious areas of low intensity within a high-intensity peripheral zone. MRI has a higher sensitivity for detecting prostate carcinomas than DRE and TRUS in patients having an elevated PSA value and a negative core biopsy. However, its specificity is poor since other abnormalities such as prostatic intraepithelial neoplasia (PIN), prostatitis, scars, or haemorrhage have a similar MRI appearance. The use of additional techniques such as MR spectroscopy and contrast-enhanced dynamic MRI improves sensitivity, but in particular it improves the specificity of tumor detection. Newly developed biopsy devices enable the performance of targeted biopsies in areas that appear suspicious in the MRI. PMID- 15902627 TI - Clinical experience with a commercially available negative oral contrast medium in PET/CT. AB - PURPOSE: To evaluate a commercially available negative oral contrast material for PET/CT. MATERIAL AND METHODS: In a prospective series of 49 patients, Mukofalk, which is a vegetarian-based substance, was used as a negative oral contrast medium in whole body PET/CT studies. Mukofalk was administered during a time period of 1.5 hours before the examination. Quality of small bowl distension and eventual pathological tracer uptake in the intestine were evaluated. RESULTS: Distension of the small bowel was excellent or good in 41 (85 %) and poor in 8 (15 %) patients. Mild tracer uptake in the small bowel was observed in 5 patients (10.2 %) and moderate uptake in another 2 patients (4 %). In none of these patients did the F-18 FDG uptake interfere with image interpretation. CONCLUSION: Mukofalk can be used as a negative oral contrast medium in PET/CT studies. PMID- 15902628 TI - The feasibility of spatial high-resolution magnetic resonance angiography (MRA) of the renal arteries at 3.0 T. AB - PURPOSE: To evaluate the feasibility of high-spatial resolution magnetic resonance angiography (MRA) of the renal arteries at 3.0 T. MATERIAL AND METHODS: Twelve healthy volunteers (mean age, 38.8 years) underwent renal MRA at 3.0 T. The application of parallel imaging with an acceleration factor of 3 allowed obtaining MR angiographic data with a voxel size of 0.9 x 0.8 x 0.9 mm in scan time of only 16 s. A dose of 0.2 mmol/kg body weight of 0.5-molar gadodiamide was administered at a flow rate of 2 ml/s. For image analysis, image quality, presence of artifacts, venous contamination and level of noise were rated by two radiologists in consensus. RESULTS: All examinations were of diagnostic quality. The image quality was rated good or very good in 91 % (11/12) of cases. Due to the high parallel imaging factor the level of noise was slightly increased without diagnostic impairment. Mild venous enhancement was found in 75 % (9/12) of the examinations. CONCLUSION: Renal MRA at 3.0 T is feasible with high spatial resolution and a short acquisition time. PMID- 15902629 TI - [Computed tomographic criteria as expected effect to inhaled nitric oxide in patients with severe acute respiratory distress syndrome]. AB - PURPOSE: Inhaled nitric oxide (iNO) is an effective therapy for severe hypoxemia in most patients with acute respiratory distress syndrome (ARDS). For unknown reason, a subset of ARDS patients does not respond favorably to iNO therapy. We hypothesized that radiological manifestation of lung injury may be related to iNO response. MATERIALS AND METHODS: We retrospectively analyzed data from n = 25 ARDS patients who received iNO, and underwent chest CT within 72 h prior to inhaled treatment. The morphology of coherently pathologic lung tissue was characterized by the length of the borderline between consolidated, infiltrated and atelectatic lung tissue and radiologically normal lung tissue. This quantity was expressed as relative fraction of the visceral pleural circumference and averaged over all CT slices. Furthermore we semiquantitatively determined the total volume of consolidated lung tissue as part of the whole lung. RESULTS: In n = 6 non-responders to iNO (DeltaPaO2 < 10 %), we determined a short relative borderline between normal and consolidated lung tissue due to the presence of large and coherently consolidated lung regions. In n = 19 iNO responders (DeltaPaO2 > 10 %), we found significantly less coherently consolidated lung tissue evidenced by an increased relative borderline when compared to iNO non responders (0.09 +/- 0.02 vs. 0.1 +/- 0.01; P < 0.05). Moreover, there was a moderate and significant correlation between DeltaPaO2 induced by iNO and the relative borderline in all patients studied (R = 0.59; P < 0.05). Total fraction of consolidated lung tissue volume was not different between iNO non-responders and responders (60 +/- 3 % vs. 54 +/- 2 % n. s.). CONCLUSION: Our data demonstrate that the gross morphological distribution of pathological lung tissue influences iNO response in ARDS. Inhaled NO was most beneficial in injured lungs characterized by many small consolidated areas surrounded by normal lung tissue. The increased borderline between pathologic and normal lung tissue offers additional possibility for iNO to divert blood flow from shunt areas to ventilated lung regions, which consequently improves arterial oxygenation. PMID- 15902630 TI - [Fluoroscopy time -- an overestimated factor for patient radiation exposure in invasive cardiology]. AB - PURPOSE: To analyze the effects of an optimized fluoroscopy time on patient radiation exposure in the course of coronary angiography (CA) and percutaneous coronary interventions (PTCA), in comparison to those with consistent collimation to the region of interest (ROI). Furthermore, to analyze efforts concerning reduction of radiographic frames as well as concerning adequate instead of best possible image quality. MATERIAL AND METHODS: For 3,115 elective CAs and 1,713 PTCA performed by one interventionist since 1997, we documented the radiographic dose-area products (DAP (R)) and fluoroscopic dose-area products (DAP (F)), the number of radiographic frames and the fluoroscopy times during selected 2-month intervals. Under conditions of constant image intensifier entrance dose, levels of DAP (R)/frame and DAP (F)/s represent valid parameters for consistent collimation. RESULTS: In 1997, the mean baseline values of DAP for elective CA and PTCA amounted to 37.1 and 31.6 Gy x cm (2), respectively. A reduction of mean fluoroscopy times from 264 to 126 seconds for CA and from 630 to 449 seconds for PCI, both resulted in an overall DAP-reduction of merely 20 %. Optimization of mean radiographic frames from 543 to 98 for CA and from 245 to 142 for PTCA enabled reductions of 53 and 13 %, respectively. By restriction to adequate instead of best-possible image quality for coronary angiography in clinical routine, we achieved an optimized radiographic DAP/frame of 30.3 to 13.3 mGy x cm (2), which enabled a 45 % reduction of overall DAP. Most efficient however was a consistent collimation to the ROI, which resulted in a remarkable radiation reduction by 46 % for CA and by 65 % for PTCA. CONCLUSIONS: Radiation-reducing educational efforts in the clinical routine of invasive cardiology should -- against widely held opinion -- focus less exclusively toward a reduction of fluoroscopy time but more efficiently toward consistent collimation to the region of interest, reduction of radiographic frames and restriction to an adequate instead of best-possible image quality. PMID- 15902631 TI - [The "EVA" Trial: Evaluation of the Efficacy of Diagnostic Methods (Mammography, Ultrasound, MRI) in the secondary and tertiary prevention of familial breast cancer. Preliminary results after the first half of the study period]. AB - PURPOSE: To investigate the respective diagnostic accuracies of the different breast imaging modalities, i. e., mammography (Mx), high-frequency breast ultrasound (US), and dynamic contrast-enhanced breast (MRI) regarding the early diagnosis of familial (hereditary) breast cancer. MATERIALS AND METHODS: A prospective, non-randomized controlled clinical multi-center trial is performed at 4 academic tertiary care centers in Germany (Ulm, Munchen/Grosshadern, Munster and Bonn) for a total period of 4 years, sponsored by the German Cancer Aid. The protocol consists of semiannual clinical visits and breast ultrasound, and annual bilateral two-view Mx, US and MRI. Imaging studies were first analyzed independently, then Mx was read in conjunction with US, followed by Mx combined with MRI, and finally, all three imaging modalities were read in synopsis. We present the concept and first results of this trial. RESULTS: So far, 748 screening rounds are available for analysis in 613 women. A total of 12 breast cancers have been identified, with 11/12 cases in the pTis or pT1/N0 stage. The mean size of detected invasive cancers was 7 mm. A total of 19 benign lesions were biopsied due to false-positive imaging diagnoses. The breast cancer detection rates were: Mx: 5/12 (42 %), US 3/12 (25 %), MRI 10/12 (83 %), and the positive predictive values: Mx 5/17 (29 %), US 3/15 (30 %), and MRI 10/23 (43 %). CONCLUSION: The preliminary data suggest that early diagnosis of familial breast cancer is feasible by intensified surveillance, in particular with the addition of MRI. PMID- 15902632 TI - [Comparison of spine injuries by means of CT and MRI according to the classification of Magerl]. AB - PURPOSE: A feasibility study of the classification of spine injuries by CT and MRI data according to the classification of Magerl for spine injuries. MATERIAL AND METHODS: In 39 patients, 46 injured vertebrae were evaluated by means of CT and MRI. A single-slice helical CT (Somatom Plus S, Siemens, Erlangen, Germany) was used with a collimation and feed of 2 mm and 3 mm, respectively. Sagittal reformations were reconstructed from these data sets. MR imaging was performed with 1.5T and 1.0T machines (Magnetom Vision and Magnetom Impact, Siemens). Transverse and sagittal T1-w SE- images as well as sagittal T2-w TSE- and STIR images were generated. Each fractured vertebra was independently categorized two times according to the classification of Magerl on the basis of CT- and MRI images. The CT- and MRI-classifications were subsequently compared. RESULTS: Most fractures were classified as impaction fractures (A1) or burst fractures (A3). Only 5/46 injuries of vertebrae were not given the same classification by means of CT and MRI (r = 0.9). MRI showed more bony lesions than CT (n. s., p > 0.05) and was able to evaluate ruptures of the longitudinal ligaments in 5 patients. CONCLUSION: Classification of spine injuries according to the Magerl classification showed a significant agreement between CT and MRI and therefore a comparable evaluation of stability. MRI was superior in delineating ligamental lesions. PMID- 15902633 TI - [Comparison of contrast-enhanced low mechanical index (Low MI) sonography and unenhanced B-mode sonography for the differentiation between synovitis and joint effusion in patients with rheumatoid arthritis]. AB - PURPOSE: To test whether contrast-enhanced low mechanical index (low MI) sonography is superior to non enhanced B-Mode sonography in differentiating synovitis and joint effusion. MATERIAL AND METHODS: In a retrospective study, 22 patients with proven rheumatoid arthritis underwent B-Mode sonography and low-MI sonography of 25 symptomatic joints of the upper and lower limbs. For low-MI sonography, 5 ml Sonovue (Bracco Altana Pharma GmbH, Konstanz) were injected as an intravenous bolus followed by 10 ml of 0.9 % saline solution. Magnetic resonance imaging (MRI) was obtained additionally in 3 joints. With non-enhanced sonography, we diagnosed a synovitis in case of an echogenic and a joint effusion in case of an anechoic mass. With contrast-enhanced sonography, we diagnosed a synovitis in case of enhancement and a joint effusion in the absence of enhancement of the intraarticular mass. RESULTS: In 13 joints, synovitis and joint effusion were differentiated by both non-enhanced and enhanced sonography. In 12 joints, this differentiation was only possible with contrast-enhanced sonography. In 3 patients diagnosed by sonography as having a synovitis, this diagnosis was proven by MRI. CONCLUSION: Contrast-enhanced low-MI sonography is superior to non-enhanced B-Mode sonography in differentiating synovitis and joint effusion. PMID- 15902634 TI - [Influence of clinical information on the diagnostic validity of MRI in the detection of abacterial sacroiliitis]. AB - PURPOSE: To evaluate the influence of clinical information on the diagnostic validity of MRI in the detection of abacterial sacroiliitis. Evaluation of sensitivities, specificities, negative (NPV) and positive predictive values (PPV) for MRI with and without clinical information in comparison to a prior defined standard. MATERIALS AND METHODS: The results of routine MRI reports of 65 patients with the clinical suspicion of abacterial sacroiliitis referred by rheumatologists for MR assessment of the sacroiliac joints were retrospectively reviewed. These results were compared to the results of a blinded reading of the same examinations performed without any clinical information. The MRI protocol included T1-SE, STIR and T1-SE contrast-enhanced sequences with fat saturation. All patients were followed for at least four years. The standard of reference was defined by an experienced rheumatologist and included baseline and follow-up data (clinical, laboratory and imaging). Abacterial sacroiliitis was diagnosed in 19 patients. RESULTS: Under routine clinical conditions, MRI revealed a sensitivity of 94.7 % and specificity of 97.8 %. A PPV of 94.7 % and a NPV of 97.8 % were achieved. Without clinical information, MRI revealed a sensitivity and specificity of 64 and 80 %, and a PPV of 49 % and a NPV of 86 %, respectively. CONCLUSION: The additional knowledge of clinical information for the MR diagnosis of abacterial sacroiliitis increases its diagnostic value for the assessment of abacterial sacroiliitis. PMID- 15902635 TI - [MRI of pulmonary arteries in follow-up after arterial-switch-operation (ASO) for transposition of great arteries (d-TGA)]. AB - PURPOSE: To determine the value of MRI in postoperative evaluation of patients with arterial-switch-operation (ASO) for d-TGA. MATERIAL AND METHODS: 14 consecutive patients with d-TGA and ASO were examined with MRI in addition to ultrasound because the acoustic window in echocardiography was insufficient. Nine patients had a pulmonary-artery-bifurcation anterior to the aortic root (group 1), and five (group 2) laterally (two of them left, three of them right). MRI was performed in six patients as contrast-enhanced MR-angiography, in 8 patients as native examination. Diameters of pulmonary arteries were measured in two perpendicular views at the origin and in two further locations each with a distance of one cm. RESULTS: One patient showed a supravalvular pulmonary artery stenosis with subsequent pulmonary artery aneurysm, which had not been detected in echocardiography. This patient showed only indirect signs of pulmonary artery stenosis in echocardiography. Compared to group 1, pulmonary arteries in group 2 generally had greater diameters. Those pulmonary arteries in Group 2 which cross the mediastinum underneath the aortic arch had lower diameters compared to the pulmonary artery positioned on the other side. In group 1, the left-sided pulmonary artery had lower diameters than the right-sided pulmonary arteries. It seems remarkable that the narrowing of pulmonary arteries is associated with a flattening of the transversal vessel diameter. CONCLUSION: Narrowing of pulmonary arteries after ASO is associated with oval vessel diameters. Therefore, multiplanar diagnostic methods are necessary. MRI meets this prerequisite and is therefore advantageous to echocardiography in follow-up examinations at patients with ASO for d-TGA. Special attention should be focused on the main pulmonary artery, on the left pulmonary artery in patients with preaortic pulmonary artery bifurcation as well as on the pulmonary artery which crosses the mediastinum in patients with laterally placed pulmonary artery bifurcation. Angiography should only be performed when intervention is planned. PMID- 15902636 TI - [Diagnostic evaluation of chronic inflammatory intestinal diseases in children and adolescents: MRI with true-FISP as new gold standard?]. AB - PURPOSE: To evaluate the impact of magnetic resonance imaging (MRI) with use of True-FISP sequences in the evaluation of inflammatory bowel-wall changes in children and adolescents with Crohn's disease. Furthermore, the diagnostic procedure in children and adolescents with chronic inflammatory bowel disease (IBD) will be discussed in light of the relevant literature. MATERIAL AND METHODS: Twenty-four children and adolescents aged between 7 and 21 years with suspected or known IBD underwent MRI on a 1.5 T-scanner (Philips ACS-NT, Best, Netherlands). One hour after 1 l of a 2.5 % mannitol solution was given orally, MR imaging was performed using coronal HASTE-M2D, coronal fat-suppressed T2-TSE, axial dynamic T1-weighted GE-sequences before and after i. v.-contrast material injection (0.1 mmol/kg Gd-DTPA) and using a 2D-balanced-FFE-sequence (True-FISP) before and after i. v.-contrast material injection in coronal and axial planes. The MR-images were correlated with endoscopy and the clinical findings. In 14 patients, a recently performed conventional radiographic enteroclysis was available. Each performed MRI sequence was evaluated by three experienced radiologists regarding the sensitivity and specificity of each sequence in the detection of inflammatory bowel wall changes. In addition, the image quality was assessed regarding the different tissue contrasts and the susceptibility to artifacts. The distension of the bowel wall and the patients' acceptance of the MRI examination were recorded. RESULTS: With a sensitivity in detecting inflammatory small bowel changes of 93.3 % (axial pre-contrast, coronal post contrast) and 100 % (axial post-contrast, coronal pre-contrast), the True-FISP outnumbers the other performed sequences (T1 = 80 %, HASTE = 13.3 % and T2-TSE = 53.3 %). The difference between True-FISP and contrast-enhanced T1 was not statistically significant, whereas the difference between True-FISP and HASTE and T2-TSE, respectively, was statistically significant. The True-FISP sequences revealed a statistically significant superiority regarding the soft-tissue differentiation in comparison to all other performed MR-sequences. The distension of the bowel wall was good in all patients. The patients' acceptance of the MRI examination was excellent. CONCLUSION: The described small bowel MRI examination is appropriate for children and adolescents. With the use of True-FISP sequences, it is a convincing method with an outstanding sensitivity in the diagnosis of IBD. Not least because of the lack of radiation exposure, small bowel MRI ought to replace conventional enteroclysis as a gold standard for IBD diagnosis in children and adolescents. PMID- 15902637 TI - [Quantification of microangiopathic lesions in brain parenchyma and age-adjusted mean scores for the diagnostic separation of normal from pathological values in senile dementia]. AB - PURPOSE: To quantify microangiopathic lesions in the cerebral white matter and to develop age-corrected cut-off values for separating normal from dementia-related pathological lesions. MATERIALS AND METHODS: In a memory clinic, 338 patients were investigated neuropsychiatrically by a psychological test battery and by MRI. Using a FLAIR sequence and a newly developed rating scale, white matter lesions (WMLs) were quantified with respect to localization, number and intensity, and these ratings were condensed into a score. The WML scores were correlated with the mini-mental state examination (MMSE) and clinical dementia rating (CDR) score in dementia patients. A non-linear smoothing procedure was used to calculate age-related mean values and confidence intervals, separate for cognitively intact subjects and dementia patients. RESULTS: The WML scores correlated highly significantly with age in cognitively intact subjects and with psychometric scores in dementia patients. Age-adjusted WML scores of cognitively intact subjects were significantly different from those of dementia patients with respect to the whole brain as well as to the frontal lobe. Mean value and confidence intervals adjusted for age significantly separated dementia patients from cognitively intact subjects over an age range of 54 through 84 years. CONCLUSION: A rating scale for the quantification of WML was validated and age adjusted mean values with their confidence intervals for a diagnostically relevant age range were developed. This allows an easy to handle, fast and reliable diagnosis of the vascular component in senile dementia. PMID- 15902638 TI - [Is the Canadian CT head rule for minor head injury applicable for patients in Germany?]. AB - PURPOSE: To evaluate the applicability of the Canadian CT Head Rule (CCHR) on head trauma patients in a German university hospital. METHODS: 122 patients (m = 74; f = 48; 40 +/- 19 years) were examined with cranial CT due to minor head trauma. The need for cranial CT according to the CCHR was evaluated retrospectively. RESULTS: With a sensitivity of 98.9 % and a specificity of 46.6 % all patients with the need for neurosurgical intervention were detected by applying the major criteria of the CCHR. Also, every patient with severe brain injury was detected by the extended criteria with a sensitivity of 99.6 % and a specificity of 34.1 %. This would have led to a reduction in the rate of cranial CT examinations by 45.1 % for the major and 22.1 % for the extended criteria. No patient with severe brain injury would have been missed by application of the criteria. CONCLUSION: The Canadian CT Head Rule for patients with minor head trauma is applicable with a very high sensitivity and the potential of significantly reducing the rate of cranial CT examinations in these patients. PMID- 15902639 TI - [CT-guided resection of pulmonary metastases]. AB - PURPOSE: To evaluate the feasibility and potential use of intraoperative computed tomography (IOP CT) as guidance for video-assisted thoracic surgery (VATS). MATERIAL AND METHODS: Fifteen consecutive patients with peripheral intrapulmonary nodules underwent a thoracoscopy with IOP CT. Solitary lesions were known in 6/15 patients (40 %, group II) whereas 9/15 (60 %, group I) patients had multiple lesions (n >/= 2). IOP CT was performed with the mobile CT scanner Philips Tomoscan M. Radiologists intraoperatively placed percutaneous marks of lung lesions after unsuccessful VATS by use of a lung marker set (Somatex, Teltow, Germany). VATS was performed under general anaesthesia and with double lumen endotracheal intubation for single lung ventilation. Imaging quality and imaging of pulmonary nodules were rated. RESULTS: IOP CT was evaluated as feasible combined with VATS. Thoracotomy was avoided in 5/15 patients where lesions could not be detected by VATS. A CT-guided biopsy was performed in two patients after an unsuccessful attempt of thoracoscopy. There were no documented side effects. CONCLUSION: First clinical results suggest that a combination of VATS and IOP CT is feasible. Thus, the number of open thoracoscopies might be decreased. Intrapulmonary lesions not detectable with VATS could be marked under CT -- guidance intraoperatively and then resected by thoracoscopy. PMID- 15902640 TI - [Percutaneous vertebroplasty for osteoporotic vertebral compression fractures: experiences and prospective clinical outcome in 26 consecutive patients with 50 vertebral fractures]. AB - PURPOSE: Prospective evaluation of vertebroplasty for osteoporotic compression fractures concerning pain reduction, demand of analgesics and quality of life. MATERIAL AND METHODS: In 26 consecutive patients, 50 vertebral fractures were treated by percutaneous vertebroplasty under fluoroscopic (n = 44) or combined fluoroscopic/CT guidance (n = 6). Prospective follow-up was performed after 6 (for 50 vertebral fractures) and 12 months (for 27 vertebral fractures). Visual analogue scale (VAS) and numeric rating scale (NRS) was applied for the assessment of pain. Subjective quality of life and analgesics demand was documented semi-quantitatively. RESULTS: Mean interval from the beginning of pain symptoms to therapy was 8.5 weeks. Vertebroplasty was technically successful in all evaluated patients. Pain severity decreased from 10 (defined at baseline as initial pain score) to 2.8 after 6 months and 2.7 after 12 months. Subjective quality of life was reported as very well, well or improved in 92 % (n = 26 after 6 months) or 100 % (n = 13 after 12 months). No need for additional analgesic therapy was observed in 69.3 % (n = 26) after 6 months and 61.5 % (n = 13) after 12 months. Eight newly developed vertebral fractures were observed during follow up, with 5 fractures directly adjacent to previously treated vertebrae. Leakage of polymethyl methacrylate (PMMA) into the intervertebral space or spinal canal was observed in 22 % and 20 % respectively (n = 50 vertebrae) . No neurologic deterioration or complications requiring surgery were observed. CONCLUSION: Vertebroplasty is a successful therapeutic approach for the treatment of osteoporotic vertebral fractures. Persistent improvement of clinical symptoms was shown at follow-up after 6 and 12 months. PMID- 15902641 TI - [Contrast media kinetic in multi-slice helical CT cannot detect rectal cancer recurrence with certainty]. AB - PURPOSE: Evaluation of different parameters of contrast media enhancement for the differentiation between scar tissue and local recurrence of rectal cancer. MATERIALS AND METHODS: We included 83 patients after operation and radiotherapy of rectal cancer. In total, 20 local recurrences were diagnosed. After administration of 75 ml Iopromide (370 mg/ml) and a delay of 65 s, the whole abdomen and pelvis were scanned in a collimation of 4 x 2.5 mm and 12.5 mm table feed per rotation. The suspected tissue was marked by the freehand ROI option in every slice and the minimum, average and maximum densities were calculated. A local recurrence was suspected if maximum density was higher than 90 HU after admission of contrast media. In addition we calculated the maximum difference of density as the difference between maximum and minimum density. RESULTS: The minimum and average densities showed no reliable differences for patients with or without local recurrence (minimum density 4 HU ( +/- 12 HU) vs. 13 HU ( +/- 21 HU), P = 0.23, average density 48 HU ( +/- 10 HU), vs. 48 HU ( +/- 17 HU)), P = 0.52. The patients suffering from local recurrence showed higher maximum densities and a higher maximum difference than the patients without recurrence (maximum density 111 HU ( +/- 13 HU) vs. 81 HU ( +/- 24 HU), P = 0.02, maximum difference 103 HU ( +/- 20 HU) vs. 76 HU ( +/- 31 HU), P = 0.06. These differences were not significant. We calculated a sensitivity of 0.6, a specificity of 0.83, a positive predictive value of 0.52 and an accuracy of 0.77. CONCLUSION: It is not possible to diagnose a recurrent rectal cancer by density values alone. PMID- 15902642 TI - [Erroneous diagnosis of liver metastasis as benign tumor in the contrast media aided "Low-MI-Real-Time" sonography with SonoVue]. PMID- 15902643 TI - [Notable bulb of jugular vein in Apert syndrome]. PMID- 15902644 TI - [Reversible intradiscal vacuum phenomenon after percutaneous kyphoplasty]. PMID- 15902645 TI - [New negative contrast medium for PET/CT]. PMID- 15902646 TI - [Flat-panel CT for estimation of in-stent restenosis]. PMID- 15902647 TI - [Precancerous conditions of the prostate -- typical ultrasound findings]. PMID- 15902648 TI - [Optimal quality radiation dosage -- in interventional cardiology]. PMID- 15902649 TI - [Intraoperative high field strength-MRI in brain tumors -- surgical procedure can be planned immediately]. PMID- 15902650 TI - [Paranasal sinus surgery -- preoperative CT lowers bleeding risk]. PMID- 15902651 TI - [Endoprostheses -- in vivo visualization of joint contacts]. PMID- 15902652 TI - [Experience -- percutaneous recanalization (PIER) in chronic ischemia of the limbs]. PMID- 15902656 TI - Mutations in PIP5K3 are associated with Francois-Neetens mouchetee fleck corneal dystrophy. AB - Francois-Neetens fleck corneal dystrophy (CFD) is a rare, autosomal dominant corneal dystrophy characterized by numerous small white flecks scattered in all layers of the stroma. Linkage analysis localized CFD to a 24-cM (18-Mb) interval of chromosome 2q35 flanked by D2S2289 and D2S126 and containing PIP5K3. PIP5K3 is a member of the phosphoinositide 3-kinase family and regulates the sorting and traffic of peripheral endosomes that contain lysosomally directed fluid phase cargo, by controlling the morphogenesis and function of multivesicular bodies. Sequencing analysis disclosed missense, frameshift, and/or protein-truncating mutations in 8 of 10 families with CFD that were studied, including 2256delA, 2274delCT, 2709C-->T (R851X), 3120C-->T (Q988X), IVS19-1G-->C, 3246G-->T (E1030X), 3270C-->T (R1038X), and 3466A-->G (K1103R). The histological and clinical characteristics of patients with CFD are consistent with biochemical studies of PIP5K3 that indicate a role in endosomal sorting. PMID- 15902657 TI - Genetic variation in the human androgen receptor gene is the major determinant of common early-onset androgenetic alopecia. AB - Androgenetic alopecia (AGA), or male-pattern baldness, is the most common form of hair loss. Its pathogenesis is androgen dependent, and genetic predisposition is the major requirement for the phenotype. We demonstrate that genetic variability in the androgen receptor gene (AR) is the cardinal prerequisite for the development of early-onset AGA, with an etiological fraction of 0.46. The investigation of a large number of genetic variants covering the AR locus suggests that a polyglycine-encoding GGN repeat in exon 1 is a plausible candidate for conferring the functional effect. The X-chromosomal location of AR stresses the importance of the maternal line in the inheritance of AGA. PMID- 15902658 TI - Acute phase proteins in dogs and cats: current knowledge and future perspectives. AB - The acute phase response is a nonspecific inflammatory reaction of the host that occurs shortly after any tissue injury. The response includes changes in the concentration of plasma proteins called acute phase proteins (APPs), some of which decrease in concentration (negative APPs), such as albumin or transferrin, and others of which increase in concentration (positive APPs), such as C-reactive protein, serum amyloid A, haptoglobin, alpha-1-acid glycoprotein, and ceruloplasmin. Most positive APPs are glycoproteins synthesized mainly by hepatocytes upon stimulation by proinflammatory cytokines and released into the bloodstream. The acute phase response and clinical application of monitoring APPs in dogs and cats are reviewed in this article, including biochemical characteristics, assays developed for each individual APP, and preanalytic and analytic factors influencing APP results that should be taken into account for proper and adequate clinical interpretation. In addition, the diagnostic use of APPs and their possible application in monitoring treatment, which can be considered one of the most interesting and promising practical applications of these proteins, will be discussed. Finally, challenges and future developments of APPs in dogs and cats will be considered, because it is expected that new and cheaper automated assays for determination of the main APPs in small animals will contribute to a wider use of these proteins as biomarkers of infection and inflammatory lesions. PMID- 15902659 TI - Fine-needle biopsy of external ear canal masses in the cat: cytologic results and histologic correlations in 27 cases. AB - BACKGROUND: The cytologic diagnosis of ear canal tumors is difficult or impossible by swab alone because cell exfoliation may be poor and neoplastic cells may be masked by associated inflammation. Fine-needle biopsy (FNB) can be used to obtain a higher yield of cells for diagnosis. OBJECTIVE: The purpose of this study was to assess the efficacy and diagnostic value of FNB and cytologic examination in providing an accurate diagnosis of masses growing in the external ear canal of cats. METHODS: Cytologic specimens from masses in the external ear canal, taken under inhaled, general anesthesia, were classified into 4 groups: 1) ceruminous gland hyperplasia or adenoma, 2) ceruminous gland adenocarcinoma, 3) inflammatory polyps, and 4) other neoplastic and non-neoplastic masses. Cytopathologic diagnoses were compared with the final histopathologic diagnoses, and indices of diagnostic test accuracy (sensitivity, specificity, likelihood ratios, diagnostic-odds ratios) were calculated. RESULTS: Twenty-seven masses (from 25 cats, including 2 cats affected bilaterally) were included in the study. The results showed good correspondence between cytologic and histologic diagnoses with an overall agreement index (kappa) of .74, a diagnostic odds ratio of 22, and 100% (27/27) agreement in the diagnosis of inflammatory polyps versus neoplasia (both benign and malignant). CONCLUSIONS: FNB cytopathology of external ear masses in the cat was sufficiently accurate for distinguishing inflammatory polyps from neoplasia. For differentiation of benign proliferation and malignant neoplasia, however, histopathologic confirmation is recommended. PMID- 15902660 TI - Use of the ADVIA 120 for differentiating extracellular and intracellular hemoglobin. AB - BACKGROUND: The use of cell-free hemoglobin (Hgb) solutions, such as Oxyglobin (Biopure Corp, Cambridge, MA, USA), as a blood substitute for the treatment of acute anemias is increasing in veterinary medicine. These solutions interfere with colorimetric tests, which do not discriminate between cellular Hgb (Hgb cell) from the patient and extracellular Hgb (Hgb-delta) from the Oxyglobin, and therefore make the monitoring of anemia, based on Hgb concentration, difficult. The ADVIA 120 hematology analyzer (Bayer Diagnostics, Tarrytown, NY, USA) evaluates Hgb by 2 methods, a standard cyanmethemoglobin colorimetric method and flow cytometry, and therefore might provide the means to differentiate extracellular and intracellular Hgb. OBJECTIVE: The objective of this study was to determine the accuracy and precision of the ADVIA 120 in differentiating extracellular from intracellular Hgb. METHODS: Anticoagulated whole blood samples from 10 healthy dogs were analyzed in triplicate on the ADVIA 120. Hgb-delta concentration was determined by adding Oxyglobin (13 g/dL) to the whole blood samples at dilutions of 1:1, 1:2, 1:4, 1:8, 1:16, and 1:32. Hgb-cell and Hgb total values were calculated and compared with actual values by linear regression. Analyses were done in triplicate and repeated 9 consecutive times to evaluate intra-assay precision of Hgb-total and Hgb-cell determinations. RESULTS: Correlation between Hgb values obtained by colorimetric (Hb-total) and flow cytometric (Hgb-cell) methods on whole blood samples was high (R(2) = .99; n = 10) with a slope of 0.96 and intercept of 0. Correlation between actual and predicted Hgb-cell values also was high (R(2) = .99), with a small positive bias (0.289 +/- 0.185; n = 60). Intra-assay precisions were high, with most coefficients of variation <2%. CONCLUSION: The ADVIA 120 is capable of differentiating Hgb-cell from Hgb-delta. The flow cytometric method is accurate and precise when compared with the cyanmethemoglobin method. A small bias between the results is unlikely to be clinically significant but may affect the ability of the ADVIA to differentiate small quantities (<0.3 g/dL) of Hgb-delta. PMID- 15902661 TI - Decreased sodium:potassium ratios in cats: 49 cases. AB - BACKGROUND: Sodium:potassium (Na:K) ratios are often reported in feline biochemical panels, although the importance of this measurement has not been investigated. OBJECTIVES: The aims of this study were to document the range of feline disease states associated with a decreased Na:K ratio, to determine the prevalence of this biochemical abnormality in a referral hospital population, and to identify any particular disease that was more likely to have a decreased Na:K ratio. METHODS: A group of 49 cats with decreased Na:K ratios was compared with a group of 50 cats with normal Na:K ratios that were randomly selected from the same hospital population. RESULTS: Twelve of the 49 cats (24.5%) had gastrointestinal disease, 10 (20.4%) had urinary disease, 8 (16.3%) had endocrine disease, 8 (16.3%) had cardiorespiratory disease, and 5 (10.0%) had diseases affecting other body systems. Six (12.2%) had artifactually decreased Na:K ratios. No cat was identified with hypoadrenocorticism. Statistical analysis revealed that, although none of these disease states was significantly over- or under-represented in the affected group, a significantly higher proportion of cats with decreased Na:K ratio had body cavity effusions (P = .025). Serum potassium concentrations were significantly higher in the affected group (P < .0001), but there was no significant difference in mean sodium concentration between the 2 groups. CONCLUSIONS: Decreased Na:K ratios frequently occur in cats with diseases other than hypoadrenocorticism, including cats with effusions. These findings should be considered when evaluating cats with this biochemical abnormality. PMID- 15902662 TI - Hematology of the sandbar shark, Carcharhinus plumbeus: standardization of complete blood count techniques for elasmobranchs. AB - BACKGROUND: Standardized hematologic methods and reference intervals have not been established for cartilaginous fishes (sharks, skates, and rays) despite the large number of animals displayed in zoos and aquariums worldwide. OBJECTIVE: The focus of this study was to validate CBC methods for sandbar shark (Carcharhinus plumbeus) blood, based on criteria established in human medicine, for the following tests: RBC count, total WBC count, PCV, hemoglobin (Hgb) concentration, and WBC differential percentages. METHODS: Replicate CBCs were performed using blood samples from 5 captive sandbar sharks. Three protocols for RBC and total WBC counts were compared, as were different centrifugation times for PCV determination, and 2 methods for Hgb concentration. Means, minimum and maximum values, and CVs were compared to CAP and CLIA performance guidelines for human tests. RESULTS: Total WBC counts in a diluent modified for elasmobranch blood, Hgb concentration by the cyanmethemoglobin method after removal of nuclei, and WBC differential percentages showed acceptable performance. PCV results were acceptable when tubes were centrifuged for at least 5 minutes. Total RBC counts by all 3 methods exceeded the acceptable error for manual counts of human cells. CONCLUSIONS: Standardized CBC tests can be used as health assessment tools for elasmobranchs. Total RBC counts should be viewed as estimates. PMID- 15902663 TI - Validation of the Nova CRT8 for the measurement of ionized magnesium in feline serum. AB - BACKGROUND: Evaluation of serum magnesium (Mg) concentration is becoming important in human and veterinary critical care medicine. An ion-selective electrode can measure the physiologically active ionized fraction. OBJECTIVES: The purpose of this study was to validate an ion-specific electrode analyzer and assay for measuring ionized Mg in feline serum and to determine a reference interval for this analyte in cats. METHODS: Venous blood samples were collected anaerobically from clinically healthy cats, and the serum was used to validate the analyzer and assay. This included investigating the stability of samples stored at different temperatures, intra- and interassay precision, linearity, analytical sensitivity, and potential interferences from bilirubin, lipemia, hemoglobin, or serum separator tubes. A reference interval was calculated. RESULTS: Serum samples evaluated for ionized Mg concentrations can be stored at 20 degrees C for < or =24 hours, at 4 degrees C for < or =72 hours, and at 20 degrees C for < or =4 weeks, when samples are minimally exposed to air. Intra- and interassay precisions had coefficients of variation (CVs) of 1.23% and 2.02%, respectively. There was good linearity using serum (r = .998; y = -0.0057 + 1.0256x) and manufacturer-supplied aqueous solutions and quality control materials (r = .999; y = 0.0110 + 0.9213x). Apparent analytical sensitivity was at least 0.015 mmol/L. Mean recovery was good for ionized Mg in samples with 1+ icterus (104%), 4+ lipemia (99.3%) and 1-4+ hemolysis (98.6%). There was no significant difference (P = .52) in ionized Mg concentrations in serum collected in tubes containing no additives compared with serum collected in glass separator tubes. The serum ionized Mg reference interval was 0.47-0.63 mmol/L (n = 40). CONCLUSIONS: The Nova CRT8 analyzer and assay provide a precise and reliable method of measuring ionized Mg concentration in feline serum. Strict adherence to sampling techniques, handling, and storage are necessary for reliable results. PMID- 15902664 TI - Usefulness of Ki-67 proliferation marker in the cytologic identification of liver tumors in dogs. AB - BACKGROUND: Performing a biopsy is currently the best method of diagnosing liver disease. To reduce possible risk factors resulting from a biopsy, liver cytology can provide an alternative technique. The diagnostic accuracy of cytology for identifying liver tumors is, however, limited. The results of cytology might be improved by using immunochemistry for Ki-67, a proliferation marker, on liver cytology specimens. OBJECTIVES: The purpose of this study was to investigate the value of Ki-67 immunochemistry on liver cytologic specimens from dogs for identifying neoplastic diseases of the liver, by comparing the results to histologic findings. METHODS: Liver biopsy and cytology samples were obtained from 30 dogs with hepatic disease. All samples were evaluated by an anatomic pathologist and a cytopathologist. Parallel Ki-67 immunochemistry of histologic and cytologic samples was performed. The gradation of Ki-67 expression in histologic and cytologic samples was assessed. RESULTS: Cytologic specimens of liver tumors (n = 9) showed <50% Ki-67-positive cells. Twenty of 21 cases of non neoplastic liver disease had no or few single Ki-67-positive cells. Using Ki-67, the diagnostic accuracy of cytologic evaluation was increased from 78% to 100% for malignant neoplasia. CONCLUSIONS: Based on the results of this study, the cytologic evaluation of liver together with Ki-67 immunochemistry can improve the diagnostic accuracy of cytology for liver neoplasia. PMID- 15902665 TI - Fractionation of canine serum magnesium. AB - BACKGROUND: Serum total magnesium (tMg) consists of 3 fractions: ionized magnesium (iMg), protein-bound magnesium (pbMg), and complexed magnesium (cMg). Serum iMg may be measured by an ion-selective electrode, but determination of pbMg and cMg has not been attempted in dogs. OBJECTIVES: The objectives of this study were to assess the validity of a micropartition system to fractionate serum tMg and to establish reference intervals for pbMg, cMg, and iMg in clinically normal dogs using this method. METHODS: Serum samples from 10 clinically healthy dogs were fractionated using a micropartition system (Centrifree YM-30, Amicon Corp, Lexington, MA, USA). Serum tMg and iMg were measured in whole serum, and tMg was also measured in the ultrafiltrate. Concentration of cMg was obtained by the subtraction of iMg from tMg concentrations of the ultrafiltrate. Protein bound Mg was calculated by subtracting the tMg concentration of the ultrafiltrate from the tMg concentration of whole serum. RESULTS: Results for pbMg and cMg using the micropartition system showed good reproducibility. Determination of tMg and iMg had acceptable inter- and intra-assay precision. Concentrations of iMg, cMg, and pbMg were 0.50 +/- 0.05 mmol/L, 0.05 +/- 0.04 mmol/L, and 0.24 +/- 0.04 mmol/L, representing 63%, 6%, and 31% of the tMg concentration, respectively. CONCLUSIONS: The micropartition system was a reproducible means to accurately assess cMg and pbMg concentrations in dogs. PMID- 15902666 TI - Morphologic characterization of specific granules in Greyhound eosinophils. AB - BACKGROUND: "Vacuolated" eosinophils (ie, eosinophils with empty, nonstaining granules) have been described previously in normal Greyhounds. However, to our knowledge, detailed studies of granules in vacuolated and normal eosinophils in this breed have not been performed. OBJECTIVE: The objective of this prospective study was to characterize some of the morphologic, ultrastructural, and cytochemical staining features of specific (primary) granules in both normal and vacuolated eosinophils in Greyhound blood. METHODS: Morphologic features of eosinophils in Wright's- and Diff-Quik-stained peripheral blood smears from 49 Greyhounds were compared with 200 blood smears from non-Greyhound dogs. Transmission electron microscopy was done on blood from 3 Greyhounds with vacuolated eosinophils and 3 with normal eosinophil granules. Blood smears from 4 of these dogs also were stained cytochemically with alkaline phosphatase (AP), chloracetate esterase (CAE), and alpha naphthyl butyrate esterase (ANBE). The morphologic features and tinctorial properties of vacuolated and normal eosinophils were compared. RESULTS: Twenty-six Greyhounds (53%) had vacuolated eosinophils and 23 (47%) had normal granulated eosinophils in smears stained with Wright's stain. Only 1% of eosinophils were vacuolated in non-Greyhound dogs. Twenty of the 23 (85%) Greyhounds with normal granulated eosinophils on Wright's stained smears had vacuolated eosinophils in smears stained with Diff-Quik. Ultrastructurally, no morphologic differences were observed between granules of vacuolated and normal eosinophils. Both vacuolated and normal eosinophils in Greyhounds were positive for AP and negative for CAE and ANBE, as expected for normal dogs. CONCLUSION: Vacuolated eosinophils in Greyhounds likely reflect, at least in part, differential staining properties of the specific granules with different hematologic stains. Ultrastuctural and cytochemical features of eosinophil granules were similar in normal and vacuolated eosinophils from Greyhounds. PMID- 15902667 TI - Serum concentration of some acute phase proteins in naturally occurring canine babesiosis: a preliminary study. AB - BACKGROUND: Serum concentrations of acute phase proteins can provide valuable diagnostic information in the detection, prognosis, or monitoring of disease. Information available on the acute phase response in naturally occurring canine babesiosis is limited. OBJECTIVE: The purpose of this investigation was to retrospectively evaluate serum concentrations of haptoglobin, C-reactive protein, and ceruloplasmin in dogs naturally infected with Babesia canis. METHODS: Haptoglobin, C-reactive protein, and ceruloplasmin concentrations were measured in serum samples from dogs with uncomplicated (n = 6) and complicated (n = 1) babesiosis and compared with 6 healthy dogs. RESULTS: Serum C-reactive protein and ceruloplasmin concentrations were significantly higher in dogs with babesiosis; however, serum haptoglobin concentration was significantly lower compared with control dogs (P <.01). CONCLUSIONS: Results of this study suggest that acute phase protein concentrations could be beneficial in the diagnosis and determination of the severity of babesiosis in dogs. PMID- 15902668 TI - Afibrinogenemia and a circulating antibody against fibrinogen in a Bichon Frise dog. AB - A 1.5-year-old female Bichon Frise dog was evaluated for a life-threatening hemorrhagic condition that occurred after ovariohysterectomy, requiring 4 whole blood transfusions. A hemostatic profile, including activated clotting time (ACT), one-stage prothrombin time (OSPT), activated partial thromboplastin time (APTT), buccal mucosal bleeding time, and specific assays (heat-precipitation microhematocrit method and electroimmunoassay) for fibrinogen, were performed to investigate the coagulopathy. Clotting times for all tests having a fibrin clot endpoint (ACT, OSPT, APTT) and buccal mucosal bleeding time were prolonged. Plasma fibrinogen was not detected by heat-precipitation microhematocrit method or electroimmunoassay. Using the Ellis-Stransky method, a mixture of patient plasma and normal canine plasma with known fibrinogen content yielded substantially less than the calculated fibrinogen concentration, indicating the presence of an interfering substance. The interferent properties of the patient's plasma were retained following heat precipitation at 56 degrees C indicating the absence of a pyroglobulin or an abnormal fibrinogen molecule. Radial immunodiffusion assay using the patient's plasma and activated thrombin confirmed the existence of an inhibitor to the formation of fibrin. Western blot analysis using the patient's plasma identified an IgG antibody that reacted with the Beta- and gamma- but not the Alpha-subunits of canine fibrinogen. Antibody was detected in samples taken 8, 16, and 68 days after the surgery; peak titers were evident at day 16. These results supported a diagnosis of afibrinogenemia with a circulating antibody inhibitor to fibrin clot formation that developed secondary to blood transfusion. PMID- 15902669 TI - Toxoplasma gondii-like organisms in skin aspirates from a cat with disseminated protozoal infection. AB - An 8-year-old, male domestic shorthair cat was referred to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania with a 3-day history of lethargy, inappetance, hyperemic skin nodules, coughing, and vomiting. Laboratory results included nonregenerative anemia, lymphopenia, thrombocytopenia, hypoalbuminemia, hyponatremia, and increased alanine aminotransferase and aspartate aminotransferase activities. Cytology of the skin nodules revealed many spindle- to crescent-shaped protozoal organisms, with morphology consistent with Toxoplasma gondii or Neospora caninum. Gross necropsy, histopathologic, immunohistochemical, and transmission electron microscopic findings confirmed a systemic protozoal infection; however, the organism exhibited characteristics of both N caninum and T gondii. Diagnosis of a T gondii-like infection was based on internal structures of the organism and positive reaction to rabbit polyclonal antibodies to T gondii. Reports of toxoplasmic dermatitis are rare in the cat and dog, and this is the first reported diagnosis of T gondii-like protozoa in skin aspirates. PMID- 15902670 TI - A "down under" lesion on the muzzle of a dog. AB - A 10-year-old, castrated, male Labrador Retriever was presented to a local veterinary practice for investigation of a firm, deeply pigmented, alopecic, subcutaneous mass (8 mm in diameter) on the left side of the muzzle. A fine needle aspirate of the mass was submitted for cytologic evaluation to the University of Florida. Microscopically, the preparation contained a predominant population of histiocytes that contained variable numbers of intracytoplasmic, negative-staining, filamentous structures consistent with Mycobacterium sp. A presumptive diagnosis of canine leproid granuloma syndrome was based on the cytologic findings and location of the lesion. Acid-fast staining revealed bright pink, acid-fast organisms within the histiocytic cells, supporting the diagnosis. The bacteria were not detected in histopathologic sections or by a polymerase chain reaction (PCR) test 1 week later, however, possibly because of spontaneous remission. Canine leproid granuloma syndrome is a common disease in Australia, but is uncommon in dogs in North America. It is caused by a novel, unnamed Mycobacterium species and usually affects the skin and subcutaneous tissues of the head and ears. A diagnosis usually can be made in Wright's-Giemsa and acid fast-stained cytologic specimens; however, definitive diagnosis requires PCR testing at a specialized laboratory. PMID- 15902671 TI - Organisms in an aspirate from an ulcerated mass: etiologic agent or mass-querade? AB - Smears prepared from a fine-needle aspirate of an ulcerated, dermal mass between the digits of a 9-month-old Bull Terrier puppy revealed a moderately cellular, mixed-cell inflammation with numerous extracellular organisms. The organisms were 3-8 microm in length, curved to oval in shape, with prominent nuclei, and (retrospectively) a rare single, narrow-based bud. The cytologic diagnosis was mycotic dermatitis due to infection with an unusual yeast. Histologic sections of the lesion showed pyogranulomatous inflammation without evidence of an infectious organism. Contamination of the cytologic smears was suspected to have taken place in the clinical pathology laboratory and was confirmed by the isolation of a yeast with identical microscopic features from a cactus located on a windowsill near the work bench. The yeast was cultured and identified as Rhodotorula minuta, an airborne contaminant and ubiquitous saprophyte. It can be difficult to differentiate external contamination from infection when interpreting a cytologic specimen, however, it is important to do so, especially for unusual or opportunistic pathogens. PMID- 15902672 TI - Conjunctival swab cytology from a guinea pig: it's elementary! AB - Three 3-month-old guinea pigs (Cavia porcellus) were evaluated for purulent ocular discharge. Conjunctival swabs were obtained for cytologic evaluation of Wright's-Giemsa-stained preparations. The specimen from the most severely affected guinea pig consisted primarily of karyolytic neutrophils and small lymphocytes. Epithelial cells occasionally were observed that contained intracytoplasmic coccoid basophilic organisms, 0.5-1.5 microm in diameter. The intraepithelial inclusions were most consistent with Chlamydia sp elementary and reticulate bodies. Specimens from the other 2 guinea pigs had a similar inflammatory response, but organisms were not observed. Polymerase chain reaction (PCR) analysis of a conjunctival swab from the most severely affected guinea pig was positive for C psittaci, which also is referred to as Chlamydophila caviae, immunotype 8, formerly known as the guinea pig inclusion conjunctivitis strain of C psittaci. Chlamydial conjunctivitis is a common problem in guinea pig populations, with C caviae being specific for this species. Cytologic identification of elementary or reticulate bodies within epithelial cells is diagnostic for the organism in Giemsa-stained preparations. However, PCR is an important complementary tool when organisms are not observed and for accurate classification of the Chlamydia species. PMID- 15902674 TI - Caring for the seriously mentally ill in Thailand: Buddhist family caregiving. AB - The purpose of this study was to explore the lived experiences from Thai Buddhist family caregivers of seriously mentally ill relatives to understand their perspectives about Buddhist caregiving. A phenomenological study of 15 Thai Buddhist family caregivers was conducted following Cohen et al.'s process for analysis. Analysis of the interviews revealed five major themes: caregiving is Buddhist belief, caregiving is compassion, caregiving is management, caregiving is acceptance, and caregiving is suffering. Although suffering from the problems posed by mental illness permeated their lives, Thai Buddhist caregivers were able to continue to maintain compassion, management, and acceptance in caregiving to their seriously mentally ill relatives. A model of Thai Buddhist caregiving, constructed from the five major themes, is presented. PMID- 15902675 TI - Exploration of the psychometric properties of an Inventory of Voice Experiences. AB - The aim of the study was to identify psychometric properties of an inventory of voice-hearing experiences (IVE) [corrected] One hundred fifteen psychiatric nurses rated the language content represented on two forms of the IVE and the Brief Psychiatric Rating Scale (BPRS) [corrected] while viewing a videotaped assessment of an experienced voice hearer. Findings revealed modest to moderate support for internal consistency and concordance of the language represented on the measures as well as moderate support for convergent validity of IVE Forms A and B and modest support for their convergence with the BPRS. Nurse ratings of the IVE [corrected] explained 14 to 15 percent [corrected] of the variation in the same nurses' ratings of the hallucinated symptom item on the BPRS measure. Other [corrected] findings from the study [corrected]augment the position that not all hallucinated voices are indicative of clinical pathology, providing an additional basis on which to further refine and test the two measures [corrected] PMID- 15902676 TI - Visual feedback therapy to enhance medication adherence in psychosis. AB - Adherence to antipsychotic medications is key to enhancing maximum responses to medication and to decreasing relapse rates among those with psychotic disorders. Visual feedback therapy used electronic monitoring to provide visual feedback of daily medication-taking behaviors, combined with supportive counseling and education components. Twenty-three participants with DSM-IV diagnoses of psychotic disorders were recruited from community and inpatient settings. No statistical significance was found for intervention weekly for 1 month, followed by monthly for 2 months, although a majority of participants either maintained or increased adherence rates at 1 month. Additional findings included associations between low insight and low adherence and descriptive data on medication adherence behaviors. PMID- 15902677 TI - The effectiveness of advanced practice registered nurses as psychotherapists. AB - This article explores the effectiveness of advanced practice registered nurses as psychotherapists. Psychiatric/mental health (PMH) clinical nurse specialists (CNSs) possess the training, skills, and abilities necessary to provide effective psychotherapy. A case study is examined involving a client with obsessive compulsive disorder who underwent psychotherapy by a PMH CNS graduate student. Evidence is presented that CNSs follow evidence-based procedures with effective outcomes. PMID- 15902678 TI - The association between negative self-descriptions and depressive symptomology: does culture make a difference? AB - Research findings that depressed Americans endorse more negative self-related adjectives than controls may be related to a shared self-enhancement cultural frame. This study examines the relationship between negative core self descriptors and depressive symptoms in 79 Japanese and 50 American women. Americans had more positive self-descriptions and core self-descriptors; however, there were no cultural group differences in number of negative self-descriptors or core self-descriptors. There was a significant correlation between negative core self-descriptor and Beck Depression Inventory (BDI) for Americans only, explaining 10.6% of the BDI variance. Analysis of variance revealed that there was significant BDI group differences for American negative core self-descriptor only. Theoretical possibilities are discussed. PMID- 15902680 TI - What is the evidence for evidence-based practice? PMID- 15902682 TI - Regulation of murine chronic colitis by CD4+CD25- programmed death-1+ T cells. AB - Naturally arising CD4(+)CD25(+) regulatory T (T(R)) cells are engaged in the maintenance of self tolerance and prevention of autoimmune diseases. However, accumulating evidence suggests that a fraction of peripheral CD4(+)CD25(-) T cells also possesses regulatory activity. Programmed death-1 (PD-1) is a new member of the CD28/CTLA-4 family, which has been implicated in the maintenance of peripheral self tolerance. Here, we identified a subpopulation of CD4(+)CD25(-)PD 1(+) T cells in the spleen of naive mice that constitutively expressed CTLA-4 and FoxP3 and was hypoproliferative in response to anti-CD3 antibody stimulation in vitro. However, the CD4(+)CD25(-)PD-1(+) T cells uniquely produced large amounts of IL-4 and IL-10 in response to anti-CD3 and anti-CD28 mAb stimulation, unlike the CD4(+)CD25(+) T(R) cells. The CD4(+)CD25(-)PD-1(+) T cells exhibited a suppressor activity against the proliferation of anti-CD3 antibody-stimulated CD4(+)CD25(-)PD-1(-) T cells in vitro, which was partially abrogated by anti-CTLA 4 mAb, but not by anti-IL-10 or anti-PD-1 mAb. Remarkably, the CD4(+)CD25(-)PD 1(+) T cells inhibited the development of colitis induced by adoptive transfer of CD4(+)CD45RB(high) T cells into C.B17-scid/scid mice, albeit to a lesser extent than CD4(+)CD25(+) T(R) cells, in a CTLA-4-dependent manner. These results indicate that the CD4(+)CD25(-)PD-1(+) T cells contain substantial amounts of T(R) cells that are involved in the maintenance of peripheral tolerance. PMID- 15902683 TI - Extracellular phosphorylation of C9 by protein kinase CK2 regulates complement mediated lysis. AB - Ecto-protein kinases (ecto-PK) are expressed on many cell types, both normal and malignant, yet their functions are largely unknown. An ecto-PK capable of phosphorylating the C9 component of the complement system is described. This C9 ecto-PK could be inhibited by TBB, Emodin and DRB, selective inhibitors of protein kinase CK2. Treatment of Raji human B lymphoma cells with these CK2 inhibitors augmented cell killing by Rituximab (anti-CD20 antibodies) and human complement. Analysis of C5b-7-bearing Raji cells showed that extracellular inhibition of the ecto-CK2 enhanced cell lysis by C8 and C9. Blocking of the membrane complement regulator CD59 with monoclonal antibodies further enhanced the effect of the CK2 inhibitors on Raji cell death by complement. C9 ecto-CK2 activity was increased on cancer cells relative to normal fibroblasts and blood cells. Therefore, ecto-CK2 appears to be an additional factor protecting cells from complement-mediated lysis, probably by phosphorylation/inhibition of complement C9. PMID- 15902684 TI - The effect of hydrodynamics-based delivery of an IL-13-Ig fusion gene for experimental autoimmune myocarditis in rats and its possible mechanism. AB - Interleukin (IL)-13 is a pleiotropic cytokine secreted by activated Th2 T lymphocytes. Th1 cytokines are assumed to exacerbate and Th2 cytokines to ameliorate rat experimental autoimmune myocarditis (EAM). Here, we examined the effect of IL-13 on EAM, using a hydrodynamics-based delivery of an IL-13-Ig fusion gene, as well as the possible mechanism of its effect. Rats were immunized on day 0, and IL-13-Ig-treated rats were injected with pCAGGS-IL-13-Ig, and control rats with pCAGGS-Ig, on day 1 or 7. On day 17, the IL-13-Ig gene therapy was effective in controlling EAM as monitored by a decreased heart weight/body weight ratio, by reduced myocarditis and by reduced atrial natriuretic peptide mRNA in the heart, as a heart failure marker. On the basis of IL-13 receptor mRNA expression in separated cells from EAM hearts, we proposed that IL-13-Ig target cells were CD11b(+) cells and non-cardiomyocytic noninflammatory (NCNI) cells, such as fibroblasts, smooth muscle or endothelial cells. IL-13-Ig inhibited expression of the genes for prostaglandin E synthase, cyclooxygenase-2, inducible nitric oxide synthase, IL-1beta and TNF-alpha in cultivated cells from EAM hearts, while it enhanced expression of the IL-1 receptor antagonist gene. We conclude that IL-13-Ig ameliorates EAM and suppose that its effectiveness may be due to the influence on these immunologic molecules in CD11b(+) and NCNI cells. PMID- 15902685 TI - Clonal dynamics of tumor-infiltrating lymphocytes. AB - The presence of tumor-infiltrating lymphocytes (TIL) provides important evidence of anti-tumor immunity in vivo. However, TIL are usually not sufficient for inhibiting tumor growth. We explored the spatial and temporal aspects of clonal accumulation of TIL using RT-PCR/single-strand conformation polymorphism analysis. In CMS5 fibrosarcomas in BALB/c mice, accumulated T cell clones were specific in that dominant TIL were identical between distant tumors. Moreover, dominant TIL in the first tumor appeared consistently in the second tumor inoculated after formation of the first tumor. These results suggest that TIL show a certain level of specific tumor surveillance. When we characterized CD4(+) and CD8(+) TIL separately, CD8(+) TIL were highly concentrated and persistently localized at the tumor site, while most CD4(+) TIL clones were less concentrated and less persistent. A functional analysis showed that TIL had a certain degree of anti-tumor activity when CD4(+) and CD8(+) TIL were co-transferred. Co transfer of CD4(+) and CD8(+) TIL exhibited equivalent anti-tumor activity, irrespective of tumor stage. However, the numbers of TIL did not increase after the early phase of tumor progression. These data suggest that TIL are specific to the tumor and potentially retain anti-tumor activity, although their accumulation in mice is impaired. PMID- 15902686 TI - Simultaneous quantification of human cytomegalovirus (HCMV)-specific CD4+ and CD8+ T cells by a novel method using monocyte-derived HCMV-infected immature dendritic cells. AB - Immature dendritic cells (DC) infected with an endotheliotropic (Huv(+)) and leukotropic (Leuk(+)) human cytomegalovirus (HCMV) strain were used as a stimulus to determine functional HCMV-specific CD4(+) and CD8(+) T cells. Infected DC were co-cultured with autologous peripheral blood mononuclear cells and both arms of T cell activation were determined by intracellular flow cytometry analysis of IFN gamma production. Efficient stimulation of HCMV-specific CD4(+) and CD8(+) T cell responses was achieved using DC productively infected with Huv(+) Leuk(+) VR1814 strain. On the contrary, a negligible CD8(+) T cell response was obtained when HCMV strains unable to infect DC, or DC pulsed with inactivated viral antigen, were used. HCMV specificity of the T cell response was confirmed in 46 HCMV seropositive and 8 HCMV-seronegative healthy subjects. A cut-off was established to discriminate between immune and nonimmune subjects. The novel ex vivo assay enables the simultaneous evaluation of HCMV-specific CD4(+) and CD8(+) T cell responses and may be a useful tool for monitoring HCMV-specific T cell activity in immunocompromised transplanted patients. PMID- 15902687 TI - Regulation of the Leishmania-induced innate inflammatory response by the protein tyrosine phosphatase SHP-1. AB - Modulation of the phagocyte protein tyrosine phosphatase (PTP) SHP-1 by the parasite Leishmania favors its survival and propagation within its mammalian host. In vivo, the absence of SHP-1 leads to virtually absent footpad swelling, accompanied by enhanced inducible nitric oxide synthase expression. In this study, using an air pouch model, we show that viable motheaten SHP-1-deficient mice harbored a stronger inflammatory response against Leishmania infection than wild-type mice. This response was portrayed by higher pro-inflammatory cytokine (TNF-alpha, IL-1beta and IL-6) expression and secretion and by greater chemokine and chemokine receptor expression. These inflammatory molecules were probably responsible for the stronger cellular recruitment, mainly of neutrophils, seen at the site of infection in viable motheaten mice within 6 h post inoculation. We also provide strong evidence that protein tyrosine phosphatases in general, and SHP-1 in particular, are important regulators of chemokine gene expression. Overall, this study suggests that the ability of Leishmania to induce SHP-1 activity in its host allows the taming of an otherwise strong innate inflammatory response that would be detrimental for its survival and progression. PMID- 15902688 TI - Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. AB - The transcription factor FOXP3 plays a key role in CD4(+)CD25(+) regulatory T cell function and represents a specific marker for these cells. Despite its strong association with regulatory T cell function, in humans little is known about the frequency of CD4(+)CD25(+) cells that express FOXP3 protein nor the distribution of these cells in vivo. Here we report the characterization of seven anti-FOXP3 monoclonal antibodies enabling the detection of endogenous human FOXP3 protein by flow cytometry and immunohistochemistry. Flow-cytometric analysis showed that FOXP3 was expressed by the majority of CD4(+)CD25(high) T cells in peripheral blood. By contrast, less than half of the CD4(+)CD25(int) population were FOXP3(+), providing an explanation for observations in human T cells that regulatory activity is enriched within the CD4(+)CD25(high) pool. Although FOXP3 expression was primarily restricted to CD4(+)CD25(+) cells, it was induced following activation of both CD4(+) and CD8(+) T cell clones. These findings indicate that the frequency of FOXP3(+) cells correlates with the level of expression of CD25 in naturally arising regulatory T cells and that FOXP3 protein is expressed by some activated CD4(+) and CD8(+) T cell clones. These reagents represent valuable research tools to further investigate FOXP3 function and are applicable for routine clinical use. PMID- 15902689 TI - New tools to identify regulatory T cells. AB - The lack of tools for direct identification of regulatory T cells (T(reg) cells) at the single-cell level has been one of the major hurdles for the study of T(reg) cells and their involvement in human disease. The identification of the transcription factor Foxp3 as a molecular correlate for T(reg) function offered the opportunity to directly identify T(reg) cells, but until recently adequate reagents were not available. The tools promising the solution for this problem have emerged through the development of transgenic mice in which Foxp3 expression drives the production of fluorescent proteins, as well as the development of mAb that are able to identify Foxp3(+) cells by histology or flow cytometry. With these new tools, the mAb in particular, it will become possible for the first time to directly probe the participation of Foxp3(+) T(reg) cells in human pathology in a quantitative fashion. PMID- 15902690 TI - Skeletal muscle involvement in human immunodeficiency virus (HIV)-infected patients in the era of highly active antiretroviral therapy (HAART). AB - Skeletal muscle involvement can occur at all stages of human immunodeficiency virus (HIV) infection, and may represent the first manifestation of the disease. Myopathies in HIV-infected patients are classified as follows: (1) HIV-associated myopathies and related conditions, including HIV polymyositis, inclusion-body myositis, nemaline myopathy, diffuse infiltrative lymphocytosis syndrome (DILS), HIV-wasting syndrome, vasculitic processes, myasthenic syndromes, and chronic fatigue; (2) muscle complications of antiretroviral therapy, including zidovudine and toxic mitochondrial myopathies related to other nucleoside-analogue reverse transcriptase inhibitors (NRTIs), HIV-associated lipodystrophy syndrome, and immune restoration syndrome related to highly active antiretroviral therapy (HAART); (3) opportunistic infections and tumor infiltrations of skeletal muscle; and (4) rhabdomyolysis. Introduction of HAART has dramatically modified the natural history of HIV disease by controlling viral replication, but, in turn, lengthening of the survival of HIV-infected individuals has been associated with an increasing prevalence of iatrogenic conditions. PMID- 15902691 TI - Muscle cramps. AB - Muscle cramps are a common problem characterized by a sudden, painful, involuntary contraction of muscle. These true cramps, which originate from peripheral nerves, may be distinguished from other muscle pain or spasm. Medical history, physical examination, and a limited laboratory screen help to determine the various causes of muscle cramps. Despite the "benign" nature of cramps, many patients find the symptom very uncomfortable. Treatment options are guided both by experience and by a limited number of therapeutic trials. Quinine sulfate is an effective medication, but the side-effect profile is worrisome, and other membrane-stabilizing drugs are probably just as effective. Patients will benefit from further studies to better define the pathophysiology of muscle cramps and to find more effective medications with fewer side-effects. PMID- 15902692 TI - Influence of culture conditions on laccase production and isozyme patterns in the white-rot fungus Trametes gallica. AB - Laccase production by the white-rot fungus Trametes gallica was studied, using twelve different media under static or shaking condition. The results indicated that organic nitrogen sources such as tryptone and peptone strongly improved laccase production. The application of an amino acid mixture and a lignin preparation also increased the formation of laccase, which was not observed in the presence of potato extract. Native polyacryl amide gel electrophoresis (PAGE) followed by laccase activity staining using guaiacol as the substrate was performed to analyze the laccase isozyme patterns under the different culture conditions employed. Zymograms revealed a total of twenty different laccase activity bands that appeared in individual patterns, dependent on the respective culture condition applied. This indicates that both the medium composition and the mode of incubation (static or shaking) influenced the laccase isozyme gene expression. This was the first time to report so many laccase isozymes in a fungus. Native PAGE with silver staining showed that laccases were the main protein productions in several media providing a potentially convenient way in purifying laccases from T. gallica. PMID- 15902693 TI - Propionibacterium acnes-induced hepatic granuloma formation is impaired in mice lacking tetraspanin CD9. AB - The granuloma is a host defence response to persistent pathogenic irritants. In the process of granuloma formation, the activation, migration, and fusion of macrophages occur locally, but the mechanisms involved remain elusive. Tetraspanins regulate cell migration and fusion by organizing functional molecular complexes in membrane microdomains. Here we investigated the role of tetraspanin CD9 in hepatic granuloma formation. Immunostaining of the liver of untreated wild-type mice showed that CD9 was expressed by vascular endothelial cells and perivenular hepatocytes. When intrahepatic granulomas were induced by intravenous injection of Propionibacterium acnes, hepatocyte CD9 was extensively upregulated, while inflammatory cells constituting granulomas were mostly negative for CD9. Compared with wild-type littermates, CD9-knockout mice showed dissemination of Propionibacterium acnes and reduced number and size of granulomas after the injection. Moreover, production of granuloma-inducing cytokines, TNF-alpha and IFN-gamma, was delayed and chemotactic activity for macrophages was suppressed in the liver of mutant mice. These results suggest that CD9 is one of the proteins that promotes granuloma formation in the liver. PMID- 15902694 TI - SEPT9_v4 expression induces morphological change, increased motility and disturbed polarity. AB - Several lines of evidence indicate that altered expression of SEPT9 is seen in human neoplasia. In particular there is evidence of altered expression of the SEPT9_v4 isoform. The functional consequences of this remain unclear. We have studied the expression of wild-type- and GTP-binding mutants (G144V and S148N) of the SEPT9_v4 isoform in the MCF7 cell line as a model for its deregulation in neoplasia. We find that SEPT9_v4 expression induces dramatic actin cytoskeletal reorganization with the formation of processes around the cell periphery. Expression of the SEPT9_v4 isoform and a G144V mutant cause delocalization of endogenous SEPT9 from filamentous structures but the S148N mutant does not have this effect. In addition SEPT9_v4 isoform expression enhances cell motility and is associated with perturbation of directional movement. Expression of SEPT9_v4 GTP binding mutants also has potent effects on morphology and motility and causes loss of normal polarity, as judged by Golgi reorientation assays. The phenotypes induced by expression of the SEPT9_v4 isoform and the GTP mutants provide an insight into possible mechanisms of SEPT9_v4 function and suggest that the GTPase functions have both ras- and rab-like features. We propose a model in which overexpression of the SEPT9_v4 isoform in neoplasia is associated with perturbation of SEPT9 complexes, leading to phenotypes associated with neoplasia. PMID- 15902695 TI - Human cytomegalovirus downregulates complement receptors (CR3, CR4) and decreases phagocytosis by macrophages. AB - Human cytomegalovirus (HCMV) infection is associated with an increased susceptibility to opportunistic infections. Although the subversion of adaptive immune responses has been extensively studied, the consequences of HCMV infection on natural immune responses are not well documented. A striking selective downmodulation of CD11b/CD18 (CR3) or CD11c/CD18 (CR4) was found upon HCMV infection, on two models, the monocytic THP-1 cell line and monocyte- derived macrophages. HCMV-infected macrophages have an altered adhesion/phagocytic capacity to Candida albicans, a pathogen responsible for some opportunistic infections in immunocompromised patients. These results suggest a new mechanism implicated in the augmentation of opportunistic infections in HCMV patients. PMID- 15902696 TI - Resistance mutations before and after tenofovir regimen failure in HIV-1 infected patients. AB - So far, no study has examined the real impact of tenofovir (TDF)-regimen failure taking into account all patients in a current clinical practice. The aim of this study was to compare the nucleoside reverse transcriptase inhibitors (NRTIs) mutation profiles observed before TDF initiation and after TDF-regimen failure. All patients with genotypic resistance tests performed in this context were selected from the database of the department of virology. The patients were categorized in two groups according to the presence (group I) or absence (group II) of thymidine analogue mutations (TAMs) documented before starting TDF. The proportions of the two groups were compared using Chi-squared tests. Odds-ratios were analyzed with a regression logistic test. Ninety-six patients met the criteria. The median number of TAMs before TDF initiation did not change at failure. The K65R mutation, absent from all baseline genotypes, developed in 19 of the 96 patients, with an incidence significantly higher in group II than in group I. In addition, five genotypes harboring K65R with TAMs or L74V mutation were observed at failure. The changes regarding the other NRTI-associated mutations concern mostly the codons 74, 75, 115, and 118. The selection of K65R was closely linked to the absence of TAMs at baseline and to the regimens sparing both protease inhibitors (PIs) and non-NRTIs. The K65R mutation can emerge even with TAMs or L74V. No obvious impact was shown on the TAMs or other NRTI mutations, although a trend towards emergence of some particular mutations was observed. PMID- 15902697 TI - Improved HIV-1 viral load determination based on reverse transcriptase activity recovered from human plasma. AB - A more sensitive version of ExaVir Load, a test that utilizes reverse transcriptase (RT) activity from virions in plasma to determine HIV-1 viral load, is described. The virions were immobilized on a gel that was washed, followed by lysis of the virions, elution of purified RT, and finally RT activity determination. The changes made to the original test were: (1) improved washing of the immobilized virions by addition of a non-lytic detergent to the wash buffer, (2) improved virion lysis procedure, including changes in salt, detergent and pH, (3) the use of larger sample volumes in the RT assay, and (4) prolonged RT reaction time. The alterations gave a tenfold increased sensitivity compared to the original version. The correlation between RT load by the current test and RNA PCR was the same as previously (r=0.90). Using colorimetric product detection, the average detection limit in a panel of 262 patient plasma from Stockholm was 0.5 fg RT/ml, corresponding to approximately 170 RNA copies/ml. None of 54 HIV-1 RNA negative samples exhibited RT. The amount of RT load positive samples were 19% for samples containing 50-400 RNA, 71% for samples with 400-1,500, and 100% among samples with >8,000 copies/ml (according to Roche Amplicor). The sensitivity could be increased further using fluorimetric detection. In conclusion, the modifications of the test described result in an important increase in sensitivity. It can now be regarded as a competitive alternative method for HIV viral load determinations. PMID- 15902698 TI - HLA-B, -DRB1/3/4/5, and -DQB1 gene polymorphisms in human immunodeficiency virus related Kaposi's sarcoma. AB - Polymorphisms of genes in the human leukocyte antigen (HLA) complex, particularly those encoding HLA-DR, have been suggested as markers of susceptibility to Kaposi's sarcoma (KS). We conducted a case-control study comparing 147 homosexual men who developed KS after infection by human immunodeficiency virus-1 (HIV-1) and human herpes virus 8 (HHV8) with 147 matched dually infected men without HIV associated KS (HIV-KS) from the Multicenter AIDS Cohort Study. HLA-B, DRB1, DRB3, DRB4, DRB5, and DQB1 polymorphisms were examined by high-resolution DNA-based methods. Differences in distributions of genetic variants were tested by conditional logistic regression. Previously reported relationships with HLA-DRB1 alleles could not be confirmed. Instead, other associations were observed. In univariate analysis, KS was weakly associated with B*2702/5 (odds ratio (OR)=0.40, 95% confidence interval (CI)=0.18-0.91). Similar or stronger associations, positive or negative, were seen for haplotypes containing class II alleles: DRB1*1302-DQB1*0604 (OR=3.67, 95% CI=1.02-13.1), DRB4 (DR53) haplotype family members [OR=0.52, 95% CI=0.32-0.85], and DRB3 (DR52) haplotype family members (OR=1.69, 95% CI=1.07-2.67). The B*1402-DRB1*0102 haplotype, which invariably contains the V281L mutation in the 21-hydroxylase gene governing adrenal steroid biosynthesis, occurred in five cases and one control (OR=5.0, 95% CI=0.58-42.8). In a final multivariable analysis, only DRB1*1302-DQB1*0604 (OR=6.43, 95% CI=1.28-32.3, P=0.02) remained significantly associated with KS. Associations of HLA-DRB families with HIV-KS could reflect underlying immune dysregulation. The HLA B*1402-DRB1*0102 haplotype associated with increased risk of KS might represent an antigen-presenting pathway unfavorable for immune response to HHV8. Alternatively, the relationship might hold a clue to the predilection of KS for men because that haplotype harbors the mutant form of the 21-hydroxylase gene. PMID- 15902699 TI - Complete genomic sequence and phylogenetic relatedness of hepatitis B virus isolates from Iran. AB - Hepatitis B virus (HBV) is one of the main etiological agents of acute and chronic liver disease that is still a major public health problem in the world. Numerous HBV isolates have grouped into eight genotypes, A to H, based on the complete genome sequence. To date, no study has been carried out on the complete HBV genome sequence in Iran. The objective of this study was to investigate the complete genome sequence organization and phylogenetic analysis of the five HBV strains, which obtained from Iranian chronic infected patients. Results showed that Iranian strains were closely related to each other, with 97-100% nucleotide similarity. Phylogenetic analysis based on the complete genome sequences and the precore/core gene sequences revealed that all strains were of genotype D, sub genotype D1 with bootstrap value 100 and 99%, respectively. The S gene encoded Arg122, Pro127, and Lys160 corresponding to subtype ayw2. Iranian HBV isolates had closely related with Turkish HBV strains. All strains had a nucleotide length of 3,182 base pair (bp) except IR-P4 strain, with a 3,185 bp in length and with a unique Phe89 insertion in the X gene. The intragenotypic divergence of the complete genome sequence of Iranian strains was 1.8% and the intergenotypic in genotype D was 3.8% and with the other genotypes was 7.9-15.4%. In conclusion, this study revealed that the HBV genotype D, sub-genotype D1, subtype ayw2 dominates in the Iranian infected patients. A single Phe89 insertion in the X gene of the one Iranian strain with an unforeseen length of 3185 bp was identified. PMID- 15902700 TI - High prevalence of hepatitis delta virus infection detectable by enzyme immunoassay among apparently healthy individuals in Mongolia. AB - A previous study revealed a high prevalence of hepatitis B surface antigen (HBsAg) and hepatitis delta virus (HDV) RNA among 249 apparently healthy individuals (mean+/-standard deviation age, 48.4+/-13.9 years; 126 males and 123 females) in Ulaanbaatar, Mongolia. To investigate further the prevalence of HDV infection there, the same serum samples obtained from the cohort were tested for the presence of immunoglobulin G (IgG) class antibody to HDV (anti-HDV) by a newly developed enzyme-linked immunosorbent assay using recombinant hepatitis delta antigen protein expressed in the pupae of silkworm as the antigen probe. Anti-HDV was detected in 42 persons (16.9%), among whom 22 (52.4%) were positive for HBsAg and 20 (47.6%) had detectable HDV RNA. Among 170 persons with anti-HBc in the absence of HBsAg, 20 (11.8%) tested positive for anti-HDV, and 1 of the 20 subjects was positive for HDV RNA. Of note, none of 55 anti-HBc-negative persons had anti-HDV, supporting the specificity of the anti-HDV assay. The optical density (OD) value of anti-HDV was significantly higher among HDV RNA-positive subjects (n=21) than among HDV RNA-negative subjects (n=21) (2.513+/-0.514 vs. 0.836+/-0.550, P<0.0001). The present study confirmed the extremely high prevalence of HDV infection in Mongolia, and identified a person who was positive for both anti-HDV and HDV RNA despite negativity for HBsAg and HBV DNA probably due to viral interference. The anti-HDV assay may be useful for further epidemiological studies on HDV infection in larger cohorts in urban and rural areas of Mongolia, where elucidation of the transmission route of HDV is required urgently. PMID- 15902701 TI - Possible risk factors for the transmission of hepatitis E virus and for the severe form of hepatitis E acquired locally in Hokkaido, Japan. AB - Hepatitis E in industrialized countries has not been well studied. To define the possible risk factors for transmission of hepatitis E virus (HEV) and for the severe form of hepatitis E in Japan, we investigated the clinical and virological characteristics of hepatitis E in 32 patients who contracted the mild (n=23) or severe form (n=9) of domestically acquired hepatitis E between 1996 and 2004 in Hokkaido, where hepatitis E is most prevalent in Japan. Nine patients with the severe form of hepatitis E included two patients with fulminant hepatitis E and seven patients who were diagnosed with severe acute hepatitis in which hepatic encephalopathy did not appear during the course of the illness despite low plasma prothrombin activity (or=20 mg/dl). At least 25 patients (78%) had consumed uncooked or undercooked pig liver and/or intestine 1-2 months before the onset of hepatitis E. When compared with the seven patients with HEV genotype 3, the 25 patients with HEV genotype 4 had a higher peak alanine aminotransferase (ALT) level (P=0.0338) and a lower level of lowest prothrombin activity (P=0.0340). The severe form of hepatitis E was associated with the presence of an underlying disease (56% [5/9] vs. 17% [4/23], P=0.0454). The study suggests that zoonotic food-borne transmission of HEV plays an important role in the occurrence of hepatitis E in Hokkaido, Japan, and that the HEV genotype and the presence of an underlying disease influence the severity of hepatitis E. PMID- 15902702 TI - Differentiation of herpes simplex virus types 1 and 2 in clinical samples by a real-time taqman PCR assay. AB - While the clinical manifestations of HSV-1 and -2 overlap, the site of CNS infection, complications, response to antivirals, frequency of antiviral resistance, and reactivation rate on mucosal surfaces varies between HSV-1 and 2. Detection of HSV DNA by PCR has been shown to be the most sensitive method for detecting HSV in clinical samples. As such, we developed a PCR-based assay to accurately distinguish HSV-1 from HSV-2. Our initial studies indicated the assay using type specific primers was slightly less efficient for detecting HSV-1 and 2 DNA than the high throughput quantitative PCR assay we utilize that employs type common primers to gB. We subsequently evaluated the type specific assay on 3,131 specimens that had HSV DNA detected in the type common PCR assay. The typing results of these specimens were compared with the monoclonal antibody staining results of culture isolates collected from the same patients at the same time, and the HSV serologic status of the patient. The typing assay accurately identified both HSV-1 and -2 with a specificity of >99.5% and was significantly more sensitive than typing by culture and subsequent monoclonal antibody assays. Complete concordance was seen between the typing assay and HSV serologic status of the patient. Dual (HSV-1 and -2) infection in clinical samples was recognized in 2.6% of clinical samples using the new typing assay. This assay, when used in combination with the type common assay, can now accurately type almost all mucosal and visceral HSV isolates by molecular techniques. PMID- 15902703 TI - Human cytomegalovirus genetic variability in strains isolated from Japanese children during 1983-2003. AB - The genetic variability of 74 human cytomegalovirus (HCMV) clinical isolates from 60 Japanese infants and children during 1983-2003 was investigated, and the relevance to their clinical course was studied. The patients consisted of 10 asymptomatic congenitally infected babies, 45 infected perinatally or postnatally resulting in HCMV mononucleosis/hepatitis and 5 immunocompromised hosts. The hypervariable region of the HCMV genome, that is the a sequence and UL144 region was analyzed using the polymerase chain reaction (PCR) and unrooted phylogenetic trees. HCMV glycoprotein B (gB) polymorphism was also studied. Unrooted phylogenetic trees of a sequence and UL144 allowed the isolates to be grouped to 5 and 3 clades, respectively. Three gB genotypes were also determined. However, there was no correlation between specific genotypes of these three genes and clinical forms, except for congenital infection which fell into one of three clades of the UL144 gene. In addition, the variability of the three genes had no correlation with each other. This implies that study of a single gene is insufficient for investigating the molecular epidemiology of HCMV. This study provides basic data on the genetic variability of HCMV in an Asian population and should help to determine the strains for vaccine candidates. PMID- 15902704 TI - Quantitative PCR in the diagnosis of CMV infection and in the monitoring of viral load during the antiviral treatment in renal transplant patients. AB - Cytomegalovirus (CMV) infection is a significant problem in transplantation. In this study, a quantitative PCR test was compared with the CMVpp65 antigenemia assay not only in the diagnosis CMV infections but especially in the monitoring of viral loads during ganciclovir treatment of CMV disease in individual renal transplant patients. Altogether 342 blood specimens were obtained from 116 patients. Blood specimens were used for Cobas Amplicor Monitor plasma PCR and for the pp65 assay. Also shell vial culture was performed. The patients with a positive pp65 finding were monitored for CMV weekly during ganciclovir treatment and/or until the antigenemia subsided. CMV was detected in 31/116 (27%) patients, of whom 14 (12%) developed CMV disease and were treated with ganciclovir. CMV was found by shell vial culture in 13/14 cases, but by PCR and pp65 test in all 14 patients. CMV was detected in 156 (45%) samples; by PCR in 121/156 (range 344 103,000 copies/ml) and by pp65 test in 138/156 (range 1-1,000 positive cells/50,000 leukocytes) and by culture in 59/156 (38%) only. The peak viral loads were significantly (P<0.0001) higher in CMV disease than in untreated infections (19,650 vs. 379 copies/ml, and 100 vs. 5pp65 positive cells). In the monitoring of individual patients, the time-related CMV-DNAemia and pp65 antigenemia correlated well during the treatment of CMV disease. In conclusion, Cobas Amplicor Monitor plasma PCR and CMVpp65 antigen assays can be equally used in the diagnosis CMV infection and in the monitoring of viral load during antiviral treatment. PMID- 15902705 TI - Islet cell related antibodies and type 1 diabetes associated with echovirus 30 epidemic: a case report. AB - Type 1 diabetes associated genes account for less than 50% of disease susceptibility. Human enteroviruses have been implicated as environmental factors that might trigger and/or accelerate this autoimmune disorder. We now report of a 12-year-old girl that developed pancreatic autoimmunity and type 1 diabetes after enteroviral infection. Diabetes-associated autoimmunity was evaluated by measurement of several islet cell related autoantibodies. Neutralizing antibodies to different enteroviruses were determined in the case and eight children suffering from aseptic meningitis during a large scale epidemic. Several types of diabetes-associated antibodies were detected post-infection in the adolescent with newly diagnosed type 1 diabetes, including islet cell antibodies (ICA) and tyrosine phosphatase antibodies (IA2A). ICA but not IA2A appeared in the non diabetic enterovirus-infected subjects. Based on virological studies, type 1 diabetes pathogenesis process could have been triggered by echovirus 30 infections. This study provides the first evidence of an association between echovirus 30 infection with the presence of pancreatic autoimmunity and type 1 diabetes. Our data suggest that echovirus 30 Cuban strain could be considered a potentially diabetogenic enteroviral variant. PMID- 15902706 TI - Alternative EBNA1 expression in organ transplant patients. AB - In order to identify patients at risk for developing post-transplant lymphoproliferative disease (PTLD), a sensitive nested RT-PCR method for detection of EBNA1 gene expression in peripheral blood cells was used. EBNA1 expression in peripheral blood samples from 60 organ recipients was analyzed and compared with 24 healthy controls in a retrospective study. Overall, EBNA1 positive samples were detected at least once in 43% of the transplant patients with post-transplant lymphoproliferative disease, in 18% of the other transplant patients and in none of the healthy controls. The odds ratio for EBNA1 expression in patients with post-transplant lymphoproliferative disease was 3.42 (95% CI=1.02-11.54) compared to other transplant recipients. Together with normal EBV Q promoter initiated EBNA1 transcripts, an alternatively spliced form was expressed in peripheral blood cells in the above-mentioned transplant patients. This transcript lacks the U leader exon in the 5'-untranslated region (UTR). We have previously identified and characterized a functional internal ribosome entry site, the EBNA IRES, in the untranslated U leader exon of EBNA1. Transfection experiments with EBNA1 coding plasmids followed by Western blot showed that the EBNA IRES promotes cap-independent translation and increases the EBNA1 protein level. The alternative EBNA1 transcript lacking this function is expressed in the majority of the investigated EBNA1-positive patient samples as well as in some EBV-positive B-cell lines. Alternative splicing in this form gives EBV potential to regulate the translation of EBNA1 by modifying the 5' UTR. These findings indicate a new mechanism for EBNA1 expression in vivo. PMID- 15902707 TI - Molecular evidence of HTLV-1 intrafamilial transmission in a non-endemic area in Argentina. AB - In the North of Argentina, an endemic area for HTLV-1, intrafamilial transmission of this virus has been observed. The HTLV-1 status in 13 family members of a seropositive blood donor from the central region of Argentina (non-endemic area) was investigated. According to serological and molecular assays, four members of this family (the blood donor, the husband, a son, and a daughter-in-law) proved to be HTLV-1 positive. LTR, tax, and env sequences from the provirus infecting the family members were identical. This strongly suggests the intrafamilial transmission of the virus. This study demonstrated intrafamilial transmission of HTLV-1 in a non-endemic area of Argentina. PMID- 15902708 TI - A simple method for the detection of measles virus genome by loop-mediated isothermal amplification (LAMP). AB - Approximately 20,000-30,000 measles patients were reported in a surveillance of infectious diseases because of low vaccine coverage of 80% in Japan. Among them, some were thought to be secondary vaccine failure (SVF) with generally mild or non-typical measles illness and sometimes became a source of further transmission. We have developed a new, sensitive, and rapid method to detect the measles virus genome by reverse transcription loop-mediated isothermal amplification (RT-LAMP). We examined 50 nasopharyngeal secretion (NPS) samples that were obtained during the 1999 outbreak and stored at -70 degrees C and fresh NPS, lymphocytes and sera from 11 patients in 2003. Total RNA was extracted from the samples and subjected to reverse transcription-polymerase chain reaction (RT PCR) and RT-LAMP. We detected the genomic RNA corresponding to at least 0.01-0.04 TCID50, 30-100 copies in samples by RT-LAMP within 60 min after extraction of RNA, and all four genotypes isolated in Japan were equally amplified. Specific DNA amplification was monitored spectrophotometrically by real time turbidimeter and the quantity of RNA was calculated. Measles virus genome was detected in 44 of 50 stored NPS by RT-PCR and in 49 by RT-LAMP. The vaccine strain was discriminated from wild strains after sequencing the LAMP products. RT-LAMP is a useful rapid diagnostic method for the detection of measles virus without any special apparatus, showing higher sensitivity than RT-PCR, and expected to be applied for hospital-based infection control and for laboratory-based measles surveillance. PMID- 15902709 TI - Dominating prevalence of P[8],G1 and P[8],G9 rotavirus strains among children admitted to hospital between 2000 and 2003 in Budapest, Hungary. AB - Group A rotaviruses are the main cause of acute dehydrating diarrhea in children, responsible for high mortality in developing countries and a significant socio economic burden associated with treating the disease in developed countries. Two rotavirus vaccine candidates predicated on either homotypic or heterotypic protection have undergone clinical trials recently and await licensure for routine use. In anticipation of a future vaccination campaign in Hungary, the diversity of rotaviruses collected from Budapest between 2000 and 2003 were analyzed by polyacrylamide gel electrophoresis (PAGE) of the viral genome and by serotyping and genotyping of the outer capsid genes, VP7 and VP4. Among 2,763 rotavirus positive specimens available for analysis, we were able to determine the electropherotype of 2,227, and, of these, 1,517 (68.1%) were subjected to G typing and 1,173 (52.7%) were subjected to P typing. We successfully G typed 1,481 (97.6%) and P typed 1,130 (96.3%) strains, respectively. A total of six G types (G1, 50.2%; G2, 2.2%; G3, 1.7%; G4, 5.8%; G6, 0.6%; and G9, 34.4%) and four P types (P[4], 3.0%; P[6], 0.7%; P[8], 89.9%; and P[9], 1.7%) were identified in nine individual combinations (P[8],G1; P[4],G2; P[8],G3; P[8],G4; P[8],G9; P[6],G4; P[4],G1; P[9],G3; and P[9],G6). The prevalence of VP7 and VP4 specificities varied from year to year. In this regard, a shift in serotype predominance from G1 in 2000-2001 (61.8%) and 2001-2002 (69.7%) to G9 in 2002 2003 (51.3%) was an intriguing observation that has been reported recently in some other countries, as well. The emergence of serotype G9 rotaviruses in Hungary and other parts of the world may have implications for future vaccine development and use, particularly, if current vaccine candidates cannot confer adequate homotypic or heterotypic protection against these strains. PMID- 15902710 TI - ORF61 protein of Varicella-zoster virus influences JNK/SAPK and p38/MAPK phosphorylation. AB - Recently, it was demonstrated that the Varicella-zoster virus (VZV) infection led to an activation of MAP kinases. The viral protein encoded by ORF61 is a major effector of JNK/SAPK and p38/MAPK phosphorylation. ORF61 shows homology to HSV-1 ICP0, a multifunctional protein that influences the activity of c-Jun in infected cells. Stable expression of ORF61 in a MeWo derived cell line gave rise to two specific effects: (i) a major decrease of VZV replication and (ii) a strongly elevated basal JNK/SAPK phosphorylation but a reduced p38/MAPK phosphorylation, which were both altered following infection. A dose-dependent inhibition of JNK/SAPK in MeWo/61 cells resulted in a step-by-step increase of VZV replication. These findings indicate (i) that ORF61 is responsible for the elevated JNK/SAPK phosphorylation and (ii) that the VZV replication and the JNK/SAPK phosphorylation are related inversely. Compared to MeWo cells, the basal phosphorylation of downstream targets c-Jun and ATF-2 was reduced following ORF61 expression but restored after infection. Subsequent cascades to induce inflammatory responses were activated insignificantly; cascades to activate apoptotic events also remained silent. These data point towards an important role of ORF61 in the fine-regulation of activation of the MAPK pathways and their downstream targets to optimize the availability of cellular factors involved in VZV gene expression. PMID- 15902711 TI - An influenza A(H3) reassortant was epidemic in Australia and New Zealand in 2003. AB - During 2003, Australia and New Zealand experienced substantial outbreaks of influenza. The strain responsible was an A(H3N2) influenza virus described as A/Fujian/411/2002-like, which had circulated as a minor variant in the previous Northern Hemisphere (NH) winter, mainly in Korea and Japan. Early in the year the isolates were very similar to those that had been previously isolated in the NH, however, a reassortant strain emerged early in the New Zealand winter, followed by the appearance of similar viruses in Australia and other regional areas. While the hemagglutinin HA1 sequence of these viruses demonstrated only minor differences from the A/Fujian/411/2002 reference strain, the neuraminidase gene was clearly different from that of other recently circulating H3 viruses and most closely matched an earlier reference strain A/Chile/6416/2001. Three internal genes (NS, NP, M) in the reassortant viruses were also more closely related to the A/Chile/6416/2001 lineage. This reassortant A(H3) virus predominated in Australia and New Zealand in 2003 was also seen in Brazil and Malaysia during 2003 and was widespread in the United States and Europe during their 2003-04 winter. Interestingly most of the strains of A(H3) that were isolated at the beginning of the 2004 winter in Australia, did not have this earlier A/Chile/6416/2001-like neuraminidase but had a neuraminidase that was similar to that of the reference strain A/Fujian/411/2002. This was suggestive of the re introduction of influenza A(H3) from other countries, however, there was still low level circulation of the reassortant virus in 2004 with isolates detected in Australia and Singapore. PMID- 15902712 TI - Correlates of HIV, HBV, and HCV infections in a prison inmate population: results from a multicentre study in Italy. AB - A cross-sectional study was undertaken on the correlates of infection for the human immunodeficiency virus (HIV) and hepatitis viruses B and C (HBV and HCV) in a sample of inmates from eight Italian prisons. A total of 973 inmates were enrolled [87.0% males, median age of 36 years, 30.4% intravenous drug users (IDUs), 0.6% men who have sex with men (MSWM)]. In this sample, high seroprevalence rates were found (HIV: 7.5%; HCV: 38.0%; anti-HBc: 52.7%; HBsAg: 6.7%). HIV and HCV seropositivity were associated strongly with intravenous drug use (OR: 5.9 for HIV; 10.5 for HCV); after excluding IDUs and male homosexuals, the HIV prevalence remained nonetheless relatively high (2.6%). HIV prevalence was higher for persons from Northern Italy and Sardinia. The age effect was U shaped for HIV and HCV infections; HBV prevalence increased with age. Tattoos were associated with HCV positivity (OR: 2.9). The number of imprisonments was associated with HIV infection, whereas the duration of imprisonment was only associated with anti-HBc. The probability of being HIV-seropositive was higher for HCV-seropositive individuals, especially if IDUs. In conclusion, a high prevalence of HIV, HCV, and HBV infections among inmates was observed: these high rates are in part attributable to the high proportion of IDUs. Frequency of imprisonment and tattoos were associated, respectively, with HIV and HCV positivity. Although it is possible that the study population is not representative of Italy's prison inmate population, the results stress the need to improve infection control measures users was prisons. PMID- 15902713 TI - Different seroprevalence and molecular epidemiology patterns of hepatitis C virus infection in Italy. AB - The epidemiological picture of hepatitis C virus (HCV) infection in the general population is largely unknown, even in developed countries. The aim of this study was to estimate the prevalence and genotype distribution of HCV amongst a large sample of the Italian general population. A total of 3,577 serum samples were collected and screened for anti-HCV antibodies. ELISA and RIBA tests were used to assess the presence of anti-HCV. NS5b region sequencing was performed for molecular characterization. Of 3,577 tested sera, 95 (2.7%) were anti-HCV positive and a genome was detected and sequenced in 50 sera. The age-adjusted prevalence was 4.4%. Seroprevalence increased with age, following a North-South gradient, and increased steeply between the 15 and 30 and 31-45 age groups. Subtype 1b showed the highest prevalence in all geographical areas and age groups, followed by subtypes 2c (detected mainly in the elderly population in Southern Italy), 4a/d, and 3a (detected exclusively in adults) and 1a. These findings confirm that Central and Southern Italy are hyperendemic areas. The high prevalence observed in adults over age 30 is mainly attributable to an increase in 1b-prevalence but also to subtypes 2c- and 3/4-infections. Age-specific prevalence data and molecular characterization of the virus suggest that two transmission patterns co-exist in Italy: one characterized by subtype 1b and 2c infections, mainly in adults older than 60 years, and the other by subtype 3 and 4 infections, mainly in the 31-60 year age group, and consistent with intravenous drug use and immigration. PMID- 15902714 TI - Genetic analysis of JC virus and BK virus from a patient with progressive multifocal leukoencephalopathy with hyper IgM syndrome. AB - A case of acute progressive multifocal leukoencephalopathy (PML) with hyper IgM syndrome 1 is reported. Viral DNA and VP1 protein of JC virus (JCV) and BK virus (BKV) were detected by immunohistochemistry, in situ hybridization, semi-nested polymerase chain (PCR) and PCR-restriction enzyme analysis. JCV DNA and VP1 protein were found in the nuclei of oligodendrocytes. The non-coding control region (NCCR) and VP1 region of the JCV genome were sequenced; this revealed a novel rearrangement pattern of the NCCR in the brain tissue. The VP1 regions of brain and urine JCV were identical and of genotype type 2A. The BKV in the urine sample was genotype I. No BKV genome was found in the brain. The novel genomic rearrangement of the JCV NCCR in the brain tissue may have altered JCV pathogenesis to induce PML; the impaired immunity from hyper IgM syndrome 1 may have enabled the rearrangement. The JCV NCCR rearrangement in the brain may have originated from the archetypal form in the urine through deletion and duplication. PMID- 15902716 TI - Synthesis of highly substituted pyridazines through alkynyl boronic ester cycloaddition reactions. PMID- 15902717 TI - Production of CETD transgenic mouse line allowing ablation of any type of specific cell population. AB - Diphtheria toxin A-chain (DT-A) is a potent inhibitor of protein synthesis. As little as a single molecule of DT-A can result in cell death. DT-A gene driven by a tissue-specific promoter is used to achieve genetic ablation of a particular cell lineage. However, this transgenic approach often results in aberrant depletion of unrelated cells. To avoid this, we established a method for specific depletion of a cell population by controlled expression of the DT-A gene via the Cre-loxP system. We produced five transgenic mice carrying CETD construct containing loxP-flanked enhanced green fluorescent protein (EGFP) cDNA and the DT A gene. Transfection of primary cultured cells derived from CETD transgenic fetus with Cre expression plasmid resulted in extensive cell loss, as expected. Bigenic (double transgenic) offspring obtained by crossbreeding between CETD and MNCE transgenic mice in which Cre expression is controlled by the myelin basic protein (MBP) promoter exhibited embryonic lethality, suggesting expression of Cre at embryonic stages. Intravenous injection of Cre expression vector to CETD mice led to generation of glomerular lesions, probably due to predominant depletion of glomerular epithelial cells. This Cre-loxP-based cell ablation technology is powerful and convenient method of generating mice lacking any chosen cell population. PMID- 15902718 TI - Roles of histamine and its receptors in allergic and inflammatory bowel diseases. AB - Mast cell has a long history of being recognized as an important mediator secreting cell in allergic diseases, and has been discovered to be involved in IBD in last two decades. Histamine is a major mediator in allergic diseases, and has multiple effects that are mediated by specific surface receptors on target cells. Four types of histamine receptors have now been recognized pharmacologically and the first three are located in the gut. The ability of histamine receptor antagonists to inhibit mast cell degranulation suggests that they might be developed as a group of mast cell stabilizers. Recently, a series of experiments with dispersed colon mast cells suggested that there should be at least two pathways in man for mast cells to amplify their own activation degranulation signals in an autocrine or paracrine manner. In a word, histamine is an important mediator in allergic diseases and IBD, its antagonists may be developed as a group of mast cell stabilizers to treat these diseases. PMID- 15902719 TI - Enhancement of humoral immune responses to HBsAg by heat shock protein gp96 and its N-terminal fragment in mice. AB - AIM: Most studies on the immune effect of gp96 were focused on its enhancement of CTLs. It is interesting to know whether gp96 could influence the humoral immune response, and whether the recombinant N-terminal fragment of gp96 could substitute native gp96 to stimulate the immune system. METHODS: gp96 isolated from livers of normal mice and its N-terminal fragment (amino acid 22-355) expressed in E coli were used for immunization of BALb/c mice. Eight groups of mice received one of the following regiments subcutaneously in 100 microL phosphate buffered saline (PBS) at an interval of 3 wk. Group 1: PBS only; group 2: gp96 only; group 3: N-terminal fragment only; group 4: HBsAg only; group 5: HBsAg+gp96; group 6: HBsAg+N-terminal fragment; group 7: HBsAg+incomplete Freud's adjuvant; group 8: HBsAg+N-terminal fragment (95 degrees heated for 30 min). Serum anti-HBsAg antibody levels were assayed by ELISA. CTL responses in splenocytes were analyzed by ELISPOT after the last vaccination. RESULTS: The average titer of serum anti-HBsAg antibody in the mice immunized with HBsAg together with gp96 or its N-terminal fragment were much higher than those immunized with HBsAg alone detected by ELISA. The cellular immune response of the mice immunized with HBsAg together with gp96 or its N-terminal fragment was not different with those immunized with HBsAg alone measured by ELISPOT assay. CONCLUSION: gp96 or its N-terminal fragment greatly improved humoral immune response induced by HBsAg, but failed to enhance the CTL response, which demonstrated the potential of using gp96 or its N-terminal fragment as a possible adjuvant to augment humoral immune response against HBV infection. PMID- 15902720 TI - Hydrophobicity of reactive site loop of SCCA1 affects its binding to hepatitis B virus. AB - AIM: To investigate the role of SCCA2 and other SCCA1 molecules in the process of hepatitis B virus (HBV) binding to mammalian cells. METHODS: SCCA1 and SCCA2 were isolated from HepG2. Binding protein (BP) genes were obtained through PCR. Recombinant baculoviruses expressing SCCA1, SCCA2, BP, and different mutants were constructed and utilized to infect mammalian cells to investigate the binding ability of infected cells to HBV. RESULTS: A SCCA1 gene (A1) was isolated from HepG2, but it appeared to lack the binding ability of infected cells to HBV. Two mutants, A1-BP and BP-A1, were constructed by interchanging the carboxyl terminal of A1 and BP. Cells expressing A1-BP showed an increased virus binding capacity, but not BP-A1. Comparison of A1 sequence with the sequence of BP indicated the presence of only three amino acid changes in the carboxyl terminal, two of them were found in the reactive site loop (RSL) of SCCA1. Primary structure assay revealed that the hydrophobicity of BP and AJ515706 in this domain was strong, but A1 was relatively weak. Changing the aa349 of A1 from low hydrophobic glutamic acid to high hydrophobic valine enhanced HBV binding. In contrast, HBV binding was reduced by changing the aa349 of BP from valine to glutamic acid. CONCLUSION: The results suggest that the hydrophobicity of RSL of SCCA1 may play an important role in HBV binding to cells. PMID- 15902721 TI - Presence and integration of HBV DNA in mouse oocytes. AB - AIM: Hepatitis B is a worldwide public health problem. To explore the feasibility of hepatitis B virus (HBV) vertical transmission via oocytes, the presence and integration of HBV DNA in mouse oocytes were studied. METHODS: Genomic DNA was isolated and metaphases were prepared, respectively from mouse oocytes cocultured with pBR322-HBV DNA plasmids. PCR, Southern blot, dot hybridization and fluorescence in situ hybridization (FISH) were performed to explore the existence and integration of HBV DNA in oocytes. RESULTS: PCR detected positive bands in the tested samples, and then Southern blot revealed clear hybridization signals in PCR products. Final washing solutions were collected for dot hybridization and no signal for HBV DNA was observed, which excluded the possibility that contamination of washing solutions gave rise to positive results of PCR and Southern blot. FISH demonstrated that 36 of 1,000 metaphases presented positive signals. CONCLUSION: HBV DNA sequences are able to pass through the zona and oolemma to enter into oocytes and to integrate into their chromosomes. HBV DNA sequences might be brought into embryo via oocytes as vectors when they are fertilized with normal spermatozoa. PMID- 15902722 TI - Multigene tracking of quasispecies in viral persistence and clearance of hepatitis C virus. AB - AIM: To investigate the evaluation of hepatitis C virus (HCV) quasispecies in the envelope region and its relationship with the outcome of acute hepatitis C. METHODS: HCV quasispecies were characterized in specimens collected every 2-6 mo from a cohort of acutely HCV-infected subjects. We evaluated two individuals who spontaneously cleared viremia and three individuals with persistent viremia by cloning 33 1-kb amplicons that spanned E1 and the 5' half of E2, including hypervariable region 1 (HVR1). To assess the quasispecies complexity and to detect variants for sequencing, 33 cloned cDNAs representing each specimen were assessed by a combined method of analysis of a single-stranded conformational polymorphism and heteroduplex analysis. The rates of both synonymous and nonsynonymous substitutions for the E1, HVR1 and E2 regions outside HVR1 were analyzed. RESULTS: Serum samples collected from chronic phase of infection had higher quasispecies complexity than those collected from acute phase of infection in all individuals examined. The genetic diversity (genetic distance) within HVR1 was consistently higher than that in the complete E1 (0.0322+/-0.0068 vs 0.0020+/-0.0014, P<0.05) and E2 regions outside HVR1 (0.0322+/-0.0068 vs 0.0017+/ 0.0011, P<0.05) in individuals with persistent viremia, but did not change markedly over time in those with clearance of viremia. For individuals with persistent viremia, the rate of nonsynonymous substitutions within the HVR1 region (2.76X10(-3)+/-1.51X10(-3)) predominated and gradually increased, as compared with that in the E1 and E2 regions outside HVR1 (0.23X10(-3)+/-0.15X10( 3), 0.50X10(-3)+/-0.10X10(-3)). By contrast, the rates of both nonsynonymous and synonymous substitutions for the E1 and E2 regions including HVR1 were consistently lower in individuals with clearance of viremia. CONCLUSION: HCV persistence is associated with a complexity quasispecies and positive selection of HVR1 by the host immune system. PMID- 15902723 TI - Generation of the regulatory protein rtTA transgenic mice. AB - AIM: To translate Tet-on system into a conditional mouse model, in which hepatitis B or C virus (HBV or HCV) gene could be spatiotemporally expressed to overcome "immune tolerance" formed during the embryonic development and "immune escape" against hepatitis virus antigen(s), an effector mouse, carrying the reverse tetracycline-responsive transcriptional activator (rtTA) gene under the tight control of liver-specific human apoE promoter, is required to be generated. METHODS: To address this end, rtTA fragment amplified by PCR was effectively inserted into the vector of pLiv.7 containing apoE promoter to create the rtTA expressing vector, i.e., pApoE-rtTA. ApoE-rtTA transgenic fragment (-6.9 kb) released from pApoE-rtTA was transferred into mice by pronucleus injection, followed by obtaining one transgene (+) founder animal from microinjection through PCR and Southern blot analysis. RESULTS: rtTA transgene which could be transmitted to subsequent generation (F1) derived from founder was expressed in a liver-specific fashion. CONCLUSION: Taken together, these findings demonstrate that rtTA transgenic mice, in which rtTA expression is appropriately targeted to the murine liver, are successfully produced, which lays a solid foundation to 'off-on-off' regulate expression of target gene (s) (e.g., HBV and/or HCV) in transgenic mice mediated by Tet-on system. PMID- 15902724 TI - Multicenter clinical study on Fuzhenghuayu capsule against liver fibrosis due to chronic hepatitis B. AB - AIM: To study the efficacy and safety of Fuzhenghuayu capsule (FZHY capsule, a capsule for strengthening body resistance to remove blood stasis) against liver fibrosis due to chronic hepatitis B. METHODS: Multicenter, randomized, double blinded and parallel control experiment was conducted in patients (aged from 18 to 65 years) with liver fibrosis due to chronic hepatitis B. Hepatic histologic changes and HBV markers were examined at wk 0 and 24 during treatment. Serologic parameters (HA, LM, P-III-P, IV-C) were determined and B ultrasound examination of the spleen and liver was performed at wk 0, 12 and 24. Liver function (liver function and serologic parameters for liver fibrosis) was observed at wk 0, 6, 12, 18 and 24. Blood and urine routine test, renal function and ECG were examined before and after treatment. RESULTS: There was no significant difference between experimental group (110 cases) and control group (106 cases) in demographic features, vital signs, course of illness, history for drug anaphylaxis and previous therapy, liver function, serologic parameters for liver fibrosis, liver histologic examination (99 cases in experimental group, 96 cases in control group), HBV markers, and renal function. According to the criteria for liver fibrosis staging, mean score of fibrotic stage(s) in experimental group after treatment (1.80) decreased significantly compared to the previous treatment (2.33, P<0.05), but there was no significant difference in mean score of fibrotic stage(s) (2.11 and 2.14 respectively). There was a significant difference in reverse rate between experimental group (52%) and control group (23.3%) in liver biopsy. With marked effect on decreasing the mean value of inflammatory activity and score of inflammation (P<0.05), Fuzhenghuayu capsule had rather good effects on inhibiting inflammatory activity and was superior to that of Heluoshugan capsule. Compared to that of pretreatment, there was a significant decrease in HA, LM, P-III-P and IV-C content in experimental group after 12 and 24 wk of treatment. The difference in HA, LM, P-III-P and IV-C content between 12 and 24 wk of treatment and pretreatment in experimental group was significantly greater than that in control group (P<0.01-0.05). The effect, defined as two of four parameters lowering more than 30% of the baseline, was 72.7% in experimental group and 27.4% in control group (P<0.01). Obvious improvement in serum Alb, ALT, AST and GGT was seen in two groups. Compared to that of control group, marked improvement in GGT and Alb was seen in experimental group (P<0.05). The effective rate of improvement in serum ALT was 72.7% in experimental group and 59.4% in control group. No significant difference was seen in blood and urine routine and ECG before and after treatment. There was also no significant difference in stable rate in ALT and serologic parameters for liver fibrosis between experimental group and control group after 12 wk of withdrawal. CONCLUSION: Fuzhenghuayu capsule has good therapeutic effects on alleviating liver fibrosis due to chronic hepatitis B without any adverse effect and is superior to that of Heluoshugan capsule. PMID- 15902725 TI - Six-year follow-up of pancreatic beta cell function in adults with latent autoimmune diabetes. AB - AIM: To investigate the characteristics of the progression of islet beta cell function in Chinese latent autoimmune diabetes in adult (LADA) patients with glutamic acid decarboxylase antibody (GAD-Ab) positivity, and to explore the prognostic factors for beta cell function. METHODS: Forty-five LADA patients with GAD-Ab positivity screened from phenotypic type 2 diabetic (T2DM) patients and 45 T2DM patients without GAD-Ab matched as controls were followed-up every 6 mo. Sixteen patients in LADA1 and T2DM1 groups respectively have been followed-up for 6 years, while 29 patients in LADA2 and T2DM2 groups respectively for only 1.5 years. GAD-Ab was determined by radioligand assay, and C-peptides (CP) by radioimmune assay. RESULTS: The percentage of patients whose fasting CP (FCP) decreased more than 50% compared with the baseline reached to 25.0% at 1.5(th) year in LADA1 group, and FCP level decreased (395.8+/-71.5 vs 572.8+/-72.3 pmol/L, P<0.05) at 2.5(th) year and continuously went down to the end of follow up. No significant changes of the above parameters were found in T2DM1 group. The average decreased percentages of FCP per year in LADA and T2DM patients were 15.8% (4.0-91.0%) and 5.2% (-3.5 to 35.5%, P=0.000) respectively. The index of GAD-Ab was negatively correlated with the FCP in LADA patients (r(s)=-0.483, P=0.000). The decreased percentage of FCP per year in LADA patients were correlated with GAD-Ab index, body mass index (BMI) and age at onset (r(s)=0.408, -0.301 and -0.523 respectively, P<0.05). Moreover, GAD-Ab was the only risk factor for predicting beta cell failure in LADA patients (B=1.455, EXP (B)=4.283, P=0.023). CONCLUSION: The decreasing rate of islet beta cell function in LADA, being highly heterogeneous, is three times that of T2DM patients. The titer of GAD-Ab is an important predictor for the progression of islet beta cell function, and age at onset and BMI could also act as the predictors. PMID- 15902726 TI - Nestin-positive progenitor cells isolated from human fetal pancreas have phenotypic markers identical to mesenchymal stem cells. AB - AIM: To isolate nestin-positive progenitor cells from human fetal pancreas and to detect their surface markers and their capability of proliferation and differentiation into pancreatic islet endocrine cells in vitro. METHODS: Islet like cell clusters (ICCs) were isolated from human fetal pancreas by using collagenase digestion. The free-floating ICCs were handpicked and cultured in a new dish. After the ICCs developed into monolayer epithelium-like cells, they were passaged and induced for differentiation. Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence stain, fluorescence-activated cell sorting (FACS) and radioimmunoassay (RIA) were used to detect the expression of cell markers. RESULTS: (1) The monolayer epithelium-like cells had highly proliferative potential and could be passaged more than 16 times in vitro; (2) RT PCR analysis and immunofluorescence stain showed that these cells expressed both nestin and ABCG2, two of stem cell markers; (3) FACS analysis revealed that CD44, CD90 and CD147 were positive, whereas CD34, CD38, CD45, CD71, CD117, CD133 and HLA-DR were negative on the nestin-positive cells; (4) RT-PCR analysis showed that the mRNA expression of insulin, glucagon and pancreatic-duodenal homeobox gene-1 was detected, whereas the expression of nestin and neurogenin 3 disappeared in these cells treated with serum-free media supplemented with the cocktail of growth factors. Furthermore, the intra-cellular insulin content was detected by RIA after the induction culture. CONCLUSION: Nestin-positive cells isolated from human fetal pancreas possess the characteristics of pancreatic progenitor cells since they have highly proliferative potential and the capability of differentiation into insulin-producing cells in vitro. Interestingly, the nestin-positive pancreatic progenitor cells share many phenotypic markers with mesenchymal stem cells derived from bone marrow. PMID- 15902727 TI - Effect of vector-expressed shRNAs on hTERT expression. AB - AIM: To study the effect of short hairpin RNAs (shRNAs) expressed from DNA vector on hTERT expression. METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expression vector pUC18U6 to form pUC18U6ht1-4, which were then introduced into HepG2 cells by using liposome mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls. hTERT mRNA in the transfected cells were quantified by using real-time fluorescent RT-PCR. RESULTS: Among the four shRNAs against hTERT, two decreased the hTERT mRNA level. Compared with the controls, pUC18U6ht which expressed the two shRNAs reduced hTERT mRNA by 39% and 49% (P<0.05). CONCLUSION: hTERT expression is inhibited by the shRNAs expressed from the DNA vector. PMID- 15902728 TI - Effects of extracellular iron concentration on calcium absorption and relationship between Ca2+ and cell apoptosis in Caco-2 cells. AB - AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relationship between the rising intracellular calcium concentration ([Ca2+]i) and cell apoptosis in human intestinal epithelial Caco-2 cells. METHODS: Primary culture was used for growing small intestinal epithelial cells. [Ca2+]i was detected by a confocal laser scanning microscope. The changes in [Ca2+]i were represented by fluorescence intensity (FI). The apoptosis was evaluated by flow cytometry. RESULTS: Isolation of epithelial cells and preservation of its three-dimensional integrity were achieved using the digestion technique of a mixture of collagenase XI and dispase I. Purification of the epithelial cells was facilitated by using a simple differential sedimentation method. The results showed that proliferation of normal gut epithelium in vitro was initially dependent upon the maintenance of structural integrity of the tissue. If 0.25% trypsin was used for digestion, the cells were severely damaged and very difficult to stick to the Petri dish for growing. The Fe3+ chelating agent desferrioxamine (100, 200 and 300 micromol/L) increased the FI of Caco-2 cells from 27.50+/-13.18 (control, n=150) to 35.71+/ 13.99 (n=150, P<0.01), 72.19+/-35.40 (n=150, P<0.01) and 211.34+/-29.03 (n=150, P<0.01) in a concentration-dependent manner. There was a significant decrease in the FI of Caco-2 cells treated by ferric ammonium citrate (FAC, a Fe3+ donor; 10, 50 and 100 micromol/L). The FI value of Caco-2 cells treated by FAC was 185.85+/ 33.77 (n=150, P<0.01), 122.73+/-58.47 (n=150, P<0.01), and 53.29+/-19.82 (n=150, P<0.01), respectively, suggesting that calcium absorption was influenced by [Fe3+]. Calcium ionophore A23187 (0.1, 1.0 and 10 micromol/L) increased the FI of Caco-2 cells from 40.45+/-13.95 (control, n=150) to 45.19+/-21.95 (n=150, P<0.01), 89.87+/-43.29 (n=150, P<0.01) and 104.64+/-51.07 (n=150, P<0.01) in a concentration-dependent manner. The positive apoptotic cell number of the Caco-2 cells after being treated with A23187 increased from 0.32% to 0.69%, 0.90% and 1.10%, indicating that the increase in the positive apoptotic cell number was positively correlated with [Ca2+]i. CONCLUSION: Ca2+ absorbability is increased with the decrease of extracellular iron concentration Fe3+ and hindered with the increase of Fe3+ consistence out of them. Furthermore, increase of [Ca2+]i can induce apoptosis in Caco-2 cells. PMID- 15902729 TI - Tetrandrine inhibits activation of rat hepatic stellate cells in vitro via transforming growth factor-beta signaling. AB - AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-beta (TGF-beta) signaling in vitro. METHODS: HSCs were isolated from rats by in situ perfusion of liver and 18% Nycodenz gradient centrifugation, and primarily cultured on uncoated plastic plates for 24 h with DMEM containing 20% fetal bovine serum (FBS/DMEM) before the culture medium was substituted with 2% FBS/DMEM for another 24 h. Then, the HSCs were cultured in 2% FBS/DMEM with tetrandrine (0.25, 0.5, 1, 2 mg/L, respectively). Cell morphological features were observed under an inverted microscope, smooth muscle-alpha-actin (alpha-SMA) was detected by immunocytochemistry and image analysis system, laminin (LN) and type III procollagen (PCIII) in supernatants were determined by radioimmunoassay. TGF-beta1 mRNA, Smad 7 mRNA and Smad 7 protein were analyzed with RT-PCR and Western blotting, respectively. RESULTS: Tetrandrine at the concentrations of 0.25-2 mg/L prevented morphological transformation of HSC from the quiescent state to the activated one, while alpha-SMA, LN and PCIII expressions were inhibited. As estimated by gray values, the expression of alpha-SMA in tetrandrine groups (0.25, 0.5, 1, 2 mg/L) was reduced from 21.3% to 42.2% (control: 0.67, tetrandrine groups: 0.82, 0.85, 0.96, or 0.96, respectively, which were statistically different from the control, P<0.01), and the difference was more significant in tetrandrine at 1 and 2 mg/L. The content of LN in supernatants was significantly decreased in tetrandrine groups to 58.5%, 69.1%, 65.8% or 60.0% that of the control respectively, and that of PCIII to 84.6%, 81.5%, 75.7% or 80.7% respectively (P<0.05 vs control), with no significant difference among tetrandrine groups. RT-PCR showed that TGF-beta1 mRNA expression was reduced by tetrandrine treatments from 56.56% to 87.90% in comparison with the control, while Smad 7 mRNA was increased 1.4-4.8 times. The TGF-beta1 mRNA and Smad 7 mRNA expression was in a significant negative correlation (r=-0.755, P<0.01), and both were significantly correlated with alpha-SMA protein expression (r=-0.938, P<0.01; r=0.938, P<0.01, respectively). The up-regulation of Smad 7 protein by tetrandrine (1 mg/L) was confirmed by Western blotting as well. CONCLUSION: Tetrandrine has a direct inhibiting effect on the activation of rat HSCs in culture. It up-regulates the expression of Smad 7 which in turn blocks TGF-beta1 expression and signaling. PMID- 15902730 TI - Cytotoxic effect of a non-peptidic small molecular inhibitor of the p53-HDM2 interaction on tumor cells. AB - AIM: To investigate if non-peptidic small molecular inhibitors of the p53-HDM2 interaction could restore p53 function and kill tumor cells. METHODS: A series of non-peptidic small HDM2 inhibitors were designed by computer-aided model and synthesized by chemical method. Syl-155 was one of these inhibitors. Cytotoxic effect of syl-155 on three tumor cell lines with various states of p53, HT1080 (wild-type p53), KYSE510 (mutant p53), MG63 (p53 deficiency) was evaluated by MTT assay, Western blot and flow cytometry. RESULTS: Syl-155 stimulated the accumulation of p53 and p21 protein in HT1080 cells expressing wild-type p53, but not in KYSE510 and MG63 cells. Consequently, syl-155 induced cell cycle arrest and apoptosis in HT1080 cells. CONCLUSION: Non-peptidic small molecular inhibitors of the p53-HDM2 interaction show promise in treatment of tumors expressing wild-type p53. PMID- 15902731 TI - Expressed genes in regenerating rat liver after partial hepatectomy. AB - AIM: To reveal the liver regeneration (LR) and its control as well as the occurrence of liver disease and to study the gene expression profiles of 551 genes after partial hepatectomy (PH) in regenerating rat livers. METHODS: Five hundred and fifty-one expressed sequence tags screened by suppression subtractive hybridization were made into an in-house cDNA microarray, and the expressive genes and their expressive profiles in regenerating rat livers were analyzed by microarray and bioinformatics. RESULTS: Three hundred of the analyzed 551 genes were up- or downregulated more than twofolds at one or more time points during LR. Most of the genes were up- or downregulated 2-5 folds, but the highest reached 90 folds of the control. One hundred and thirty-nine of them showed upregulation, 135 displayed downregulation, and up or down expression of 26 genes revealed a dependence on regenerating livers. The genes expressed in 24-h regenerating livers were much more than those in the others. Cluster analysis and generalization analysis showed that there were at least six distinct temporal patterns of gene expression in the regenerating livers, that is, genes were expressed in the immediate early phase, early phase, intermediate phase, early late phase, late phase, terminal phase. CONCLUSION: In LR, the number of down regulated genes was almost similar to that of the upregulated genes; the successively altered genes were more than the rapidly transient genes. The temporal patterns of gene expression were similar 2 and 4 h, 12 and 16 h, 48 and 96 h, 72 and 144 h after PH. Microarray combined with suppressive subtractive hybridization can effectively identify the genes related to LR. PMID- 15902732 TI - Effects of emodin and double blood supplies on liver regeneration of reduced size graft liver in rat model. AB - AIM: To study the influences of emodin and reconstruction of double blood supplies on liver regeneration of reduced size graft liver in rat model. METHODS: A total of 45 SD-SD rat reduced size liver transplantation models were randomly divided into three groups (A-C). The conventional reduced size liver transplantation was performed on rats in group A, while the hepatic artery blood supply was restored in groups B and C. The emodin (1.5 mg/kg/d) was given by intraperitoneal route in group C only. The recipients were killed on the seventh day after the operation. The proliferative cell nuclear antigen (PCNA), TBil and ALT of serum were detected, and the pathological changes of liver cell were observed. RESULTS: The numbers of the rats that survived in A, B, and C group on the seventh day after operation were 14, 13, 13, respectively. The levels of TBil (31.5+/-5.2 micromol/L, 23.2+/-3.1 micromol/L vs 38.6+/-6.8 micromol/L), and ALT (5 351+/-1 050 nKat, 1300+/-900 nKat vs 5779+/-1202 nKat) in serum in groups B and C were lower than those in group A (P<0.05), while the expression of PCNA in groups B or C was higher than that in group A (22.0+/-3.5%, 28.2+/-4.2% vs 18.6+/ 3.2%, P<0.05). The deeper staining nuclei, double nuclei, multi-nuclei and much glycogen were observed in liver cells of groups B and C, especially in group C, while fewer were found in liver cells of group A. CONCLUSION: The reconstruction of arterial blood supply is very important for rat liver regeneration after reduced size liver transplantation. Emodin has the effect of promoting liver regeneration and improving liver function in rats after reduced size transplantation. The possible mechanism is improving proliferation of liver cell and protecting liver cells from injury. PMID- 15902733 TI - Diagnosis of biliary strictures after liver transplantation: which is the best tool? AB - AIM: To evaluate the diagnostic value of different indirect methods like biochemical parameters, ultrasound (US) analysis, CT-scan and MRI/MRCP in comparison with endoscopic retrograde cholangiography (ERC), for diagnosis of biliary complications after liver transplantation. METHODS: In 75 patients after liver transplantation, who received ERC due to suspected biliary complications, the result of the cholangiography was compared to the results of indirect imaging methods performed prior to ERC. The cholangiography showed no biliary stenosis (NoST) in 25 patients, AST in 27 and ITBL in 23 patients. RESULTS: Biliary congestion as a result of AST was detected with a sensitivity of 68.4% in US analysis (specificity 91%), of 71% in MRI (specificity 25%) and of 40% in CT (specificity 57.1%). In ITBL, biliary congestion was detected with a sensitivity of 58.8% in the US, 88.9% in MRI and of 83.3% in CT. However, as anastomotic or ischemic stenoses were the underlying cause of biliary congestion, the sensitivity of detection was very low. In MRI detected the dominant stenosis at a correct localization in 22% and CT in 10%, while US failed completely. The biochemical parameters, showed no significant difference in bilirubin (median 5.7; 4,1; 2.5 mg/dL), alkaline phosphatase (median 360; 339; 527 U/L) or gamma glutamyl transferase (median 277; 220; 239 U/L) levels between NoST, AST and ITBL. CONCLUSION: Our data confirm that indirect imaging methods to date cannot replace direct cholangiography for diagnosis of post transplant biliary stenoses. However MRI may have the potential to complement or precede imaging by cholangiography. Optimized MRCP-processing might further improve the diagnostic impact of this method. PMID- 15902734 TI - Expression of bcl-2 protein in chronic hepatitis C: effect of interferon alpha 2b with ribavirin therapy. AB - AIM: Mechanisms responsible for persistence of HCV infection and liver damage in chronic hepatitis C are not clear. Apoptosis is an important form of host immune response against viral infections. Anti-apoptotic protein bcl-2 expression on liver tissue as well as the influence of interferon alpha 2b (IFNalpha2b) and ribavirin (RBV) were analyzed in patients with chronic hepatitis C. METHODS: In 30 patients with chronic hepatitis C (responders--R and non-responders--NR) treated with IFNalpha2b+RBV, protein bcl-2 was determined in hepatocytes and in liver associated lymphocytes before and after the treatment. RESULTS: The treatment diminished bcl-2 protein accumulation in liver cells in patients with hepatitis C (P<0.05). Before and after the therapy, we detected bcl-2 protein in R in 87+/-15% and 83+/-20% of hepatocytes and in 28+/-18% and 26+/-10% of liver associated lymphocytes, respectively. In NR, the values before treatment decreased from 94+/-32% to 88+/-21% of hepatocytes and 39+/-29% to 28+/-12% of lymphocytes with bcl-2 expression. There was no statistical correlation between bcl-2 expression on liver tissue with inflammatory activity, fibrosis and biochemical parameters before and after the treatment. CONCLUSION: IFNalpha2b+RBV treatment, by bcl-2 protein expression decrease, enables apoptosis of hepatocytes and associated liver lymphocytes, which in turn eliminate hepatitis C viruses. PMID- 15902735 TI - Computed tomography findings in fatal cases of enormous hepatic portal venous gas. AB - AIM: To assess the computed tomography (CT) findings in the patients with hepatic portal venous gas (HPVG) who presented with a short fatal clinical course in our hospital in order to demonstrate if there was any sign for prediction. METHODS: Between January 1997 and December 2000, CT scan of the abdomen was performed on 949 patients with acute abdominal pain in our emergency department. Five patients were found having HPVG. The CT images and clinical presentations of all these five patients were reviewed. RESULTS: In reviewing the CT findings of the cases, HPVG in bilateral hepatic lobes, abnormal gas in the superior mesenteric veins, small bowel intramural gas, and bowel distension were observed in all patients. Dry gas in multiple branches of the mesenteric vein was also revealed in all cases. All the patients expired due to irreversible septic shock within 48 h after their initial clinical presentation in emergency room. Two patients had acute pancreatitis with grade D and E Balthazar classification and they expired within 24 h due to progressing septic shock under aggressive medical treatment and life support. Two patients with underlying end stage renal disease expired within 48 h even though emergent surgical intervention was undertaken. The excited bowels revealed severe ischemic change. One patient expired only a few hours after the CT examination. CONCLUSION: HPVG is a diagnostic clue in patients with acute abdominal conditions, and CT is the most specific diagnostic tool for its evaluation. The dry mesenteric veins are the suggestive fatal sign, especially for the deteriorating patients, with the direct effect on gastrointestinal perfusion. PMID- 15902736 TI - Relationship between proliferative activity of cancer cells and clinicopathological factors in patients with esophageal squamous cell carcinoma. AB - AIM: To assess whether the molecular markers of malignant tumors could improve the understanding of tumor characteristics, and to observe the characteristics of expression of cell cycle markers Ki-67 and cyclin A in esophageal carcinoma and to analyze the relationship between proliferative activity of cancer cells and clinicopathological factors. METHODS: Seventy of surgically resected esophageal squamous cell carcinoma (SCC) were examined by immunohistochemistry utilizing commercially available antibodies. Nuclear staining was regarded as a positive result. At least 50 fields in each tumor and non-tumor section were evaluated at a medium power (X200) to determine the proportion of tumor cells and the staining intensity of nuclei in the entire sections. RESULTS: Ki-67 and cyclin A were only expressed in base cells of normal esophageal mucosa. The positive immuno-staining of nuclei of SCC was significantly higher than that in normal esophageal mucosa (t=13.32 and t=7.52, respectively, P<0.01). The distribution of positively stained was more diffuse and stronger in poorly differentiated SCC. Both Ki-67 and cyclin A expressions were related to histological grades of tumors (t=3.5675 and t=3.916; t=2.13, respectively, P<0.05) but not to the sex and age of the patients, tumor size, lymphatic invasion, location, or stage grouping. CONCLUSION: The proliferative activity of cancer cells may be understood by immunohistochemistry of Ki-67 and cyclin A in Chinese patients with esophageal SCC. These cell cycle markers may serve as an indicator of cancer cell proliferation rate. The overexpression of cell cycle markers Ki-67 and cyclin A suggests the poor SCC differentiation in patients with esophageal carcinoma. PMID- 15902737 TI - Phage displaying peptides mimic schistosoma antigenic epitopes selected by rat natural antibodies and protective immunity induced by their immunization in mice. AB - AIM: To obtain the short peptides mimic antigenic epitopes selected by rat natural antibodies to schistosomes, and to explore their immunoprotection against schistosomiasis in mice. METHODS: Adults worm antigens (AWA) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme linked transferred immunoblotting methods with normal SD rat sera (NRS). The killing effects on schistosomula with fresh and heat-inactivated sera from SD rats were observed. Then the purified IgG from sera of SD rats was used to biopan a phage random peptide library and 20 randomly selected positive clones were detected by ELISA and 2 of them were sequenced. Sixty female mice were immunized thrice with positive phage clones (0, 2nd), 4th wk). Each mouse was challenged with 40 cercariae, and all mice were killed 42 d after challenge. The worms and the liver eggs were counted. RESULTS: NRS could specifically react to the molecules of 75,000, 47,000, 34,500 and 23,000 of AWA. Sera from SD rats showed that the mortality rate of schistosomula was 76.2%, and when the sera were heat inactivated in vitro, the mortality rate was decreased to 41.0% after being cultured for 48 h. The specific phages bound to IgG were enriched about 300-folds after three rounds of biopanning. Twenty clones were detected by ELISA, 19 of them bound to the specific IgG of rat sera. Immunization with these epitopes was carried out in mice. Compared with the control groups, the mixture of two mimic peptides could induce 34.9% (P=0.000) worm reduction and 67.6% (P=0.000) total liver egg reduction in mice. Two different mimic peptides could respectively induce 31.0% (P=0.001), 14.5% (P=0.074) worm reduction and 61.2% (P=0.000), 35.7% (P=0.000) total liver egg reduction. The specific antibody could be induced by immunization of the mimic peptides, and the antibody titer in immunized mice reached more than 1:6,400 as detected by ELISA. CONCLUSION: Specific peptides mimic antigenic molecules can be obtained by biopanning the phage random peptide library and a partially protective immunity against schistosome infection can be stimulated by these phage epitopes in mice. PMID- 15902738 TI - Chinese medicine compound Changtong oral liquid on postoperative intestinal adhesions. AB - AIM: The aim of this study was to observe the effect of a Chinese medicine compound Changtong oral liquid (CT) on tissue plasminogen activity (t-PA), plasminogen activator inhibitor (PAI), TGF-beta1 and hydroxyproline (OHP). METHODS: Two sets of animal experiments were performed in the present study. Forty New Zealand rabbits and 48 Sprague-Dawley (SD) rats were assigned randomly to one of the five groups: sham adhesion, adhesion with saline, adhesion with low dosage of the CT, adhesion with middle dosage of the CT and adhesion with high dosage of the CT. t-PA and PAI activity in plasma, OHP and TGF-beta1 expression in adhesion were investigated. Analysis of variance was used to test differences among groups. RESULTS: CT treatment increased plasma t-PA activity in rabbits but decreased TGF-beta1 activity in rats. The data were expressed from low to high dose respectively as follows: t-PA, 46.1+/-8.6 microkat/L, 59.6+/-10.1 microkat/L, 64.0+/-11.5 microkat/L; TGF-beta1 28+/-7.23%, 31+/-3.05%, 30+/-4.04%. There were significant differences compared with saline-treated animals (t-PA 26.4+/-5.1 microkat/L, TGF-beta1 54+/-5.51%). OHP content in cecum of rabbits from middle and high but not low dose of CT lowered significantly as compared with saline-treated rabbits, 0.3641+/-0.1373, 0.3348+/-0.0321, 0.2757+/-0.0497 mg/g vs 0.4183+/-0.0883 mg/g of protein, P>0.05, P<0.05, P<0.05 respectively. The rabbit plasma PAI activity and OHP content in abdominal wall had no difference in all groups. CONCLUSION: CT treatment significantly enhanced t-PA activity in rabbits, but decreased TGF-beta1 content in rats, OHP content in cecum of rabbits, and failed to affect the activity of PAI and OHP content in abdominal wall in rabbits, compared with saline group. The result suggests that CT could effectively prevent adhesions without interfering wound healing. PMID- 15902739 TI - Clinical features of probable severe acute respiratory syndrome in Beijing. AB - AIM: To summarize clinical features of probable severe acute respiratory syndrome (SARS) in Beijing. METHODS: Retrospective cases involving 801 patients admitted to hospitals in Beijing between March and June 2003, with a diagnosis of probable SARS, moderate type. The series of clinical manifestation, laboratory and radiograph data obtained from 801 cases were analyzed. RESULTS: One to three days after the onset of SARS, the major clinical symptoms were fever (in 88.14% of patients), fatigue, headache, myalgia, arthralgia (25-36%), etc. The counts of WBC (in 22.56% of patients) lymphocyte (70.25%) and CD3, CD4, CD8 positive T cells (70%) decreased. From 4-7 d, the unspecific symptoms became weak; however, the rates of low respiratory tract symptoms, such as cough (24.18%), sputum production (14.26%), chest distress (21.04%) and shortness of breath (9.23%) increased, so did the abnormal rates on chest radiograph or CT. The low counts of WBC, lymphocyte and CD3, CD4, CD8 positive T cells touched bottom. From 8 to 16 d, the patients presented progressive cough (29.96%), sputum production (13.09%), chest distress (29.96%) and shortness of breath (35.34%). All patients had infiltrates on chest radiograph or CT, some even with multi-infiltrates. Two weeks later, patients' respiratory symptoms started to alleviate, the infiltrates on the lung began to absorb gradually, the counts of WBC, lymphocyte and CD3, CD4, CD8 positive T cells were restored to normality. CONCLUSION: The data reported here provide evidence that the course of SARS could be divided into four stages, namely the initial stage, progressive stage, fastigium and convalescent stage. PMID- 15902740 TI - Syndecan-1 and E-cadherin expression in differentiated type of early gastric cancer. AB - AIM: To elucidate the role and alterations of syndecan-1 and E-cadherin expression in different cellular phenotypes of differentiated-type gastric cancers (DGCs). METHODS: A total of 120 DGCs at an early stage, and their adjacent mucosa, were studied both by immunohistochemistry. Syndecan-1 and E cadherin were assessed by immunohistochemical staining with anti-syndecan-1 and anti-E-cadherin antibodies, respectively. Based on immunohistochemistry, DGCs and their surrounding mucosa were divided into four types: gastric type (G-type), ordinary type (O-type), complete-intestinal type (CI-type), and null type (N type). RESULTS: Syndecan-1 expression was significantly lower in G-type cancers (29.4%) than in O-type (79.6%) and CI-type cancers (90%) (P<0.05, respectively), but E-cadherin did not show this result. In addition, syndecan-1 expression was significantly reduced in DGCs comprised partly of poorly differentiated adenocarcinoma or signet-ring cell carcinoma, compared to DGCs demonstrating papillary and/or tubular adenocarcinoma (P<0.05). G-type intestinal metaplasia (IM) surrounding the tumors was observed in 23.8% of G-type, 4.9% of O-type, and 6.7% of CI-type cancers (P<0.05; G-type vs O-type). Reduction of syndecan-1 expression was significant in G-type IM (25%) compared to non-G-type IM (75%; P<0.05). CONCLUSION: Loss of syndecan-1 plays a role in the growth of G-type cancers of DGCs at an early stage, and the reduction of syndecan-1 expression in IM surrounding the tumors may influence the growth of G-type cancer. PMID- 15902741 TI - Combined human growth hormone and lactulose for prevention and treatment of multiple organ dysfunction in patients with severe chronic hepatitis B. AB - AIM: To evaluate the efficiency and safety of combined recombinant human growth hormone (rhGH) and lactulose for treatment and/or prevention of multiple organ dysfunction in patients with chronic severe hepatitis B. METHODS: Forty-eight inpatients with chronic severe hepatitis B were randomly divided into rhGH group (n=28) and control group (n=20). In rhGH group, 4-4.5 IU of rhGH was injected intramuscularly once daily for 2-4 wk, and 100 mL of enema containing 30 mL of lactulose, 2 g of metronidazole and 0.9% saline was administered every 2 d for 2 4 wk. Their symptoms and complications were noted. Liver and kidney functions were analyzed by an Olympus analyzer. Serum GH, IGF-1, IGFBP1 and IGFBP3 were measured by ELISA. RESULTS: Clinical symptoms of 90% of these patients in rhGH group were obviously improved. The total effectiveness in rhGH group was better than that in control group (75% vs 40%, P<0.05). After 2- and 4-wk treatment of rhGH respectively, serum albumin (26.1+/-4.1 vs 30.2+/-5.3, 31.9+/-5.1 g/L), prealbumin (79.6+/-28.0 vs 106.6+/-54.4, 108.4+/-55.0 g/L), cholesterol (76.3+/ 16.7 vs 85.6+/-32.3, 96.1+/-38.7 mg/dL), and IGFBP1 (56.8+/-47.2 vs 89.7+/-50.3 ng/mL after 2 wk) were significantly increased compared to control group (P<0.05). However, serum GH was decreased. The increase of serum IGF1 and IGFBP3 after rhGH treatment was also observed. CONCLUSION: rhGH in combination with lactulose may be beneficial to the prevention and treatment of multiple organ dysfunction in patients with chronic severe hepatitis. PMID- 15902742 TI - Protective effect of fufanghuangqiduogan against acute liver injury in mice. AB - AIM: To study the effects and possible mechanisms of fufanghuangqiduogan (FFHQ) in mice with acute liver injury (ALI). METHODS: ALI was successfully induced by injecting carbon tetrachloride (CCl4) intraperitoneally and by tail vein injection of Bacillus Calmette Guerin (BCG) and lipopolysaccharide (LPS) in mice, respectively. Each of the two model groups was divided into normal group, model group, FFHQ (60, 120 and 240 mg/kg) treatment groups, and bifendate treatment group. At the end of the experiment, levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), content of malondialdehyde (MDA), activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) in liver homogenate were measured by biochemical methods. The activities of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) were determined by radio immunoassay. Hepatic tissue sections were stained with hematoxylin and eosin and examined under a light microscope. RESULTS: In the two models of ALI, FFHQ (60, 120, 240 mg/kg) was found to significantly decrease the serum transaminase (ALT, AST) activities. Meanwhile, FFHQ decreased MDA contents and upregulated the lower SOD and GSH-px levels in liver homogenate. Furthermore, in immunologic liver injury model, FFHQ decreased levels of TNF-alpha and IL-1 in serum. Histologic examination showed that FFHQ could attenuate the area and extent of necrosis, reduce the immigration of inflammatory cells. CONCLUSION: FFHQ had protective effect on liver injury induced by either CCl4 or BCG+LPS in mice, and its mechanisms were related to free radical scavenging, increasing SOD and GSH-px activities and inhibiting the production of proinflammatory mediators. PMID- 15902743 TI - Docetaxel shows radiosensitization in human hepatocellular carcinoma cells. AB - AIM: To determine the radiosensitizing potential of docetaxel in human hepatocellular carcinoma SMMC-7721 cells and its mechanisms. METHODS: SMMC-7721 cells were incubated with docetaxel at 0.125, 0.25, and 0.5 nmoL/L for 24 h and at 0.125 and 0.25 nmol/L for 48 h before irradiation. Radiation doses were given from 0 to 10 Gy. Cell survival was measured by a standard clonogenic assay after a 9-d incubation. The reactive oxygen species (ROS) and glutathione (GSH) are detected after being given the same dose of docetaxel for the same time. RESULTS: The sensitization enhancement ratios (SER) for SMMC-7721 cells determined at the 50% survival level were 1.15, 1.21 and 1.49 at 0.125, 0.25, and 0.5 nmol/L for pre-incubation of 24 h, respectively; the SER were 1.42, 1.67 at 0.125 and 0.25 nmol/L, for pre-incubation of 48 h, respectively. The ROS of SMMC-7721 cells increased and GSH decreased after pretreatment with the same doses of docetaxel for 24 or 48 h. CONCLUSION: A radiosensitizing effect of docetaxel could be demonstrated unambiguously in this cell line used. In addition, our data showed that the mechanism of radiopotentiation by docetaxel probably does not involve a G2/M block in SMMC-7721 cells, and ROS generation and GSH deletion may play a key role in the radiosensitizing effect of docetaxel. PMID- 15902744 TI - Protective effect of L-arginine preconditioning on ischemia and reperfusion injury associated with rat small bowel transplantation. AB - AIM: To investigate the protective effect and possible mechanism of L-arginine preconditioning on ischemia and reperfusion injury associated with small bowel transplantation (SBT). METHODS: Male inbred Wistar rats weighting between 180 and 250 g were used as donors and recipients in the study. Heterotopic rat SBT was performed according to the techniques of Li and Wu. During the experiment, intestinal grafts were preserved in 4 degrees Ringer's solution for 8 h before being transplanted. Animals were divided into three groups. In group 1, donors received intravenous L-arginine (50 mg/kg, 1 mL) injection 90 min before graft harvesting. However, donors in control group were given normal saline (NS) instead. In group 3, six rats were used as sham-operated control. Specimens were taken from intestinal grafts 15 min after reperfusion. Histological grading, tissue malondialdehyde (MDA) and myeloperoxidase (MPO) levels were assessed. The graft survival of each group was monitored daily until 14 d after transplantation. RESULTS: Levels of MDA and MPO in intestine of sham-operated rats were 2.0+/-0.22 mmol/g and 0.66+/-0.105 U/g. Eight hours of cold preservation followed by 15 min of reperfusion resulted in significant increases in tissue MDA and MPO levels. Pretreatment with L-arginine before graft harvesting resulted in lower enhancement of tissue levels of MDA and MPO and the differences were significant (4.71+/-1.02 mmol/g vs 8.02+/-3.49 mmol/g, 1.03+/ 0.095 U/g vs 1.53+/-0.068 U/g, P<0.05). Besides, animals in L-arginine pretreated group had better histological structures and higher 2-wk graft survival rates comparing with that in NS treated group (3.3+/-0.52 vs 6+/-0.1, 0/6 vs 6/6, P<0.05 or 0.01). CONCLUSION: L-arginine preconditioning attenuates ischemia and reperfusion injury in the rat SBT model, which was due to antioxidant activities partially. PMID- 15902745 TI - Mutations of p53 gene exons 4-8 in human esophageal cancer. AB - AIM: To characterize the tumor suppressor gene p53 mutations in exon 4, esophageal cancer and adjacent non-cancerous tissues. METHODS: We performed p53 (exons 4-8) gene mutation analysis on 24 surgically resected human esophageal cancer specimens by PCR, single-strand conformation polymorphism, and DNA sequencing. RESULTS: p53 gene mutations were detected in 9 of 22 (40.9%) esophageal cancer specimens and 10 of 17 (58.8%) adjacent non-cancerous tissues. Eight of sixteen (50.0%) point mutations detected were G-A transitions and 9 of 18 (50.0%) p53 gene mutations occurred in exon 4 in esophageal cancer specimens. Only 1 of 11 mutations detected was G-A transition and 4 of 11 (36.4%) p53 gene mutations occurred in exon 4 in adjacent non-cancerous tissues. CONCLUSION: Mutation of p53 gene in exon 4 may play an important role in development of esophageal cancer. The observation of p53 gene mutation in adjacent non-cancerous tissues suggests that p53 gene mutation may be an early event in esophageal carcinogenesis. Some clinical factors, including age, sex, pre-operation therapy and location of tumors, do not influence p53 gene mutation rates. PMID- 15902746 TI - Recurrent acute pancreatitis and its relative factors. AB - AIM: To evaluate the causes and the relative factors of recurrent acute pancreatitis. METHODS: From 1997 to 2000, acute pancreatitis relapsed in 77 of 245 acute pancreatitis patients. By reviewing the clinical treatment results and the follow-up data, we analyzed the recurrent factors of acute pancreatitis using univariate analysis and multivariate analysis. RESULTS: Of the 245 acute pancreatitis patients, 77 were patients with recurrent acute pancreatitis. Of them, 56 patients relapsed two times, 19 relapsed three times, each patient relapsed three and four times. Forty-seven patients relapsed in hospital and the other 30 patients relapsed after discharge. Eighteen patients relapsed in 1 year, eight relapsed in 1-3 years, and four relapsed after 3 years. There were 48 cases of biliary pancreatitis, 3 of alcohol pancreatitis, 5 of hyperlipidemia pancreatitis, 21 of idiopathic pancreatitis. Univariate analysis showed that the patients with local complications of pancreas, obstructive jaundice and hepatic function injury were easy to recur during the treatment period of acute pancreatitis (P=0.022<0.05, P=0.012<0.05 and P=0.002<0.05, respectively). Multivariate analysis showed that there was no single factor related to recurrence. Of the 47 patients who had recurrence in hospital, 16 had recurrence in a fast period, 31 after refeeding. CONCLUSION: Acute pancreatitis is easy to recur even during treatment. The factors such as changes of pancreas structure and uncontrolled systemic inflammatory reaction are responsible for the recurrence of acute pancreatitis. Early refeeding increases the recurrence of acute pancreatitis. Defining the etiology is essential for reducing the recurrence of acute pancreatitis. PMID- 15902747 TI - Mild hypothermia protects liver against ischemia and reperfusion injury. AB - AIM: To determine whether mild hypothermia could protect liver against ischemia and reperfusion injury in pigs. METHODS: Twenty-four healthy pigs were randomly divided into normothermia, mild hypothermia and normal control groups. The experimental procedure consisted of temporary interruption of blood flow to total hepatic lobe for different lengths of time and subsequent reperfusion. Hepatic tissue oxygen pressure (PtiO2) and aspartate aminotransferase (AST) values were evaluated, and ultrastructural analysis was carried out for all samples. RESULTS: Serum AST was significantly lower, and hepatic PtiO2 values were significantly higher in the mild hypothermia group than in the normothermia group during liver ischemia-reperfusion periods (P=0.032, P=0.028). Meanwhile, the histopathologic injury of liver induced by ischemia-reperfusion was significantly improved in the mild hypothermia group, compared with that in the normothermia group. CONCLUSION: Mild hypothermia can protect the liver from ischemia-reperfusion injury in pigs. PMID- 15902749 TI - Abstracts of the 37th Annual Meeting of the American Burn Association. May 10-13, 2005, Chicago, Illinois, USA. PMID- 15902748 TI - The right hepatic artery syndrome. AB - Various benign and malignant conditions could cause biliary obstruction. Compression of extrahepatic bile duct (EBD) by right hepatic artery was reported as a right hepatic artery syndrome but all cases were compressed EBD from stomach side. Our case compressed from dorsum was not yet reported, so it was thought to be a very rare case. We present here the first case of bile duct obstruction due to the compression of EBD from dorsum by right hepatic artery. PMID- 15902751 TI - Bibliography. Current world literature. Rheumatoid arthritis. PMID- 15902750 TI - [Recommendations for the clinical practice of early discharge after delivery: conditions for proposing early homecoming (May 2004)]. PMID- 15902752 TI - Bibliography. Current world literature. Crystal deposition diseases. PMID- 15902753 TI - [Orthopedic refresher. Adhesive capsulitis. Etiology, diagnosis and therapy]. PMID- 15902754 TI - Abstracts of the 24th Annual Meeting of the American Pain Society. March 30-April 2, 2005. PMID- 15902755 TI - [Conference review. Colon cancer: what treatment in 2004? The point in five questions]. PMID- 15902756 TI - [Abecedary of the "Annales". Part 3]. PMID- 15902757 TI - Judson J. Van Wyk, M.D.: his life and legacy. PMID- 15902758 TI - The creation of a patient-classification system in an outpatient infusion center setting. PMID- 15902759 TI - Current trends in neuropathic pain treatments with special reference to fibromyalgia. AB - Neuropathic pain and fibromyalgia are prevalent diseases which have major consequences on healthcare resources and the individual. From the clinical point of view neuropathic pains represent a heterogeneous group of aetiologically different diseases ranging from cancer to diabetes. Patients with fibromyalgia also display clinical features common in neuropathic pain suggesting that there might be some overlap. The mechanisms responsible for symptoms and signs in both diseases are still unknown. Recently, there have been numerous reports of various pharmacological treatments of neuropathic pain and fibromyalgia with often disappointing results. Most of the studies were of short duration, had high attrition rates, and displayed other methodological problems. Some compounds had high rates of adverse effects which makes it often difficult for the patients to tolerate the treatment, especially in the long-term. At present, the best options for medication treatment are tricyclic antidepressants in lower dosage than usual in psychiatric disorders and a wide range of anticonvulsants. Opioids are sometimes recommended but have been found to have minor efficacy. Recently, there have been more controlled trials, which are reported here if they had been published between 2002 and 2004. Various compounds have been tested in different studies. Treatment of fibromyalgia, which has many features in common with depressive symptoms, became the focus of interest. New promising studies with dual serotonin-norepinephrine reuptake inhibitors (duloxetine and milnacipram) and a newer antiepileptic drug (pregabalin) are in progress. Future research will have to apply new approaches (e.g., using a mechanism-based classification of neuropathic pain and carrying out studies in populations with the same symptom but not necessarily the same disease) in order to find effective treatments for these common and often debilitating diseases. PMID- 15902760 TI - Ask us. Some orthodontists claim they can complete treatment relatively quickly, and they rarely treat patients as long as 3 years. How can they do this and still achieve high-quality results? PMID- 15902761 TI - Ethnicity and suicidality. PMID- 15902762 TI - Risk reduction studies in schizophrenia. PMID- 15902763 TI - Patient handout. Does constipation ruin your day? What you eat, drink, and do can make a difference. PMID- 15902765 TI - Third Tysabri adverse case hits drug class. PMID- 15902766 TI - Mergers in Japan help firms retain own products. PMID- 15902767 TI - FDA pharmacogenomics guidance sends clear message to industry. PMID- 15902768 TI - An array of problems. PMID- 15902769 TI - Interferences: rights of the first inventor in the US. PMID- 15902770 TI - Robert Langer. PMID- 15902771 TI - Type 2 diabetes market. PMID- 15902772 TI - Clofarabine. PMID- 15902773 TI - WHO publishes definitive atlas on global heart disease and stroke epidemic. PMID- 15902774 TI - WHO issues revised drinking water guidelines to help prevent water-related outbreaks and disease. PMID- 15902776 TI - Suicide huge but preventable public health problem, says WHO. PMID- 15902775 TI - Combining TB treatment with HIV testing and treatment could save lives of up to 500,000 HIV-positive Africans every year. PMID- 15902777 TI - World Health Organization launches new initiative to address health needs of a rapidly ageing population. PMID- 15902778 TI - World facing "silent emergency" as billions struggle without clean water or basic sanitation, say WHO and UNICEF. PMID- 15902779 TI - Guideline at-a-glance. ACG-revised GERD guidelines focus on proper use of drugs. PMID- 15902780 TI - Prognostic value of early response to treatment combined with conventional risk factors in pediatric acute lymphoblastic leukemia. AB - To determine useful prognostic factors in treating childhood acute lymphoblastic leukemia (ALL), we correlated conventional risk factors and bone marrow response 14 days after induction chemotherapy. Our study included 116 precursor B-cell (n = 104) and T-cell (n = 12) ALL patients treated with our protocol between 1988 and 1999. The patients were classified into 3 initial risk groups on the basis of conventional risk factors (56 in the low-risk, 33 in the high-risk, and 27 in the very high-risk groups). All patients received similar systemic chemotherapy regimens before the evaluation of their bone marrow on day 14. We evaluated the marrow of 69 patients as M1 (less than 5% blasts), 25 as M2 (5%-25% blasts), and 22 as M3 (more than 25% blasts). Although all patients attained an initial complete remission (CR), relapse was noted in 33 of the 116 patients, and 15 patients died. All of the M1 marrow patients, irrespective of the initial risk group, showed the best event-free survival rate (85.1% +/- 3 4.4%), the lowest relapse rate (14.5%), and the highest attainment of a second CR (100%); they were defined as the new R1 prognostic group. The low-risk patients with M2 or M3 marrow (R2 group) had a relatively high relapse rate, but all of these relapsed patients were treated successfully with subsequent therapy. High- or very high risk patients with M2 or M3 marrow (R3 group) had the worst prognosis. Our new prognostic definition (R1, R2, R3) incorporating day 14 marrow findings is useful to tailor early-phase treatments for better therapeutic results in childhood ALL. PMID- 15902781 TI - [Nose, nasal sinuses, midface, orbits, hypophysis. I]. PMID- 15902782 TI - [Quality assurance study of rectal cancer]. PMID- 15902783 TI - [Communication of the DIVI]. PMID- 15902784 TI - Quality of structured abstracts of original research articles in the British Medical Journal, the Canadian Medical Association Journal and the Journal of the American Medical Association: a 10-year follow-up study. AB - BACKGROUND: We compared the quality of structured abstracts of original research articles from the British Medical Journal (BMJ), Canadian Medical Association Journal (CMAJ), and the Journal of the American Medical Association (JAMA) from 1991 to 1992 and 2001 to 2002 between journals. METHODS: A random, stratified sample of 54 abstracts from 2001 to 2002 in BMJ, CMAJ, and JAMA was compiled and coded. Two blinded raters reviewed 27 abstracts each against 33 objective criteria, separated into eight categories (purpose, research design, setting, subjects, intervention, measurement of variables, results, and conclusion). The quality score was the proportion of criteria present (range = 0-1). RESULTS: The overall mean quality score (0.74) for 2001-2002 was significantly higher than the 1988-1989 unstructured abstracts (mean = 0.57; p<0.001) but not different from the 1991-1992 structured abstracts (mean = 0.74; p>0.05). In 2001-2002, abstracts of CMAJ and JAMA (both means = 0.76) improved significantly over 1991-1992 (p<0.05) and scored significantly higher than BMJ (mean = 0.71; d.f. = 16, p<0.05). Some individual criteria scores (intervention, statistical information) improved but information was found consistently under-represented in areas that imply shortcomings of the studies. INTERPRETATION: We found a consistency in abstract quality regardless of the precise format used by different journals. This indicates that the framework for research articles already in place should be maintained and further modification of the framework may not necessarily improve the abstract quality. PMID- 15902785 TI - [Relation between cardiovascular diseases and obstructive sleep apnea syndrome]. PMID- 15902786 TI - Amiodarone fails to improve survival in amitriptyline-poisoned mice. AB - OBJECTIVE: Amiodarone, a class III antidysrhythmic agent, blocks Na+, Ca2+, and K+ channels as well as the beta-adrenergic receptor. Despite increased use of amiodarone for wide-complex tachycardia, its efficacy in the treatment of dysrhythmias induced by tricyclic antidepressants has not been tested. We investigated the effect of amiodarone and amitriptyline in a mouse lethality model. METHODS: The LD50 of amitriptyline obtained from reference sources was confirmed by giving 100 mg/kg to 40 mice by intraperitoneal (IP) injection. The safety of the treatment dose of amiodarone was confirmed by giving 50 mg/kg by IP injection to 10 mice. One hundred and nine mice were randomized to receive pretreatment with 50 mg/kg amiodarone (n=55) or an equal volume of saline or water as a volume control (n=54). Thirty minutes after pretreatment or control injection, the mice received amitriptyline, 100 mg/kg. Outcome was defined as death or survival 3 h after amitriptyline injection. RESULTS: In our confirmation of the LD50 of amitriptyline, 25/40 mice died (62.5%). None of the 10 mice that received only amiodarone died. In the control + amitriptyline arm, 36/54 (66.7%) died, compared with 39/55 (70.9%) in the amiodarone+amitriptyline arm (X2, p=0.663). Power analysis demonstrated a 90% chance of finding a 28% difference. CONCLUSIONS: Pretreatment with amiodarone does not appear to significantly alter the lethality of amitriptyline poisoning in mice. Given the inability to monitor cardiac activity in this model, further investigation in a larger animal is required. PMID- 15902787 TI - Atypical experience: a case series of pediatric aripiprazole exposures. AB - BACKGROUND: Aripiprazole is a new psychotropic agent that possesses a unique pharmacologic profile. The drug demonstrates mixed dopamine and serotonin agonist antagonist activity and has been labeled a third-generation antipsychotic and dopamine-serotonin system stabilizer. Overdose experience is limited, especially in pediatrics. CASE SERIES: Of five pediatric cases identified, toxicity was mainly evident in younger patients. A 2-year-old who ingested 40 mg experienced vomiting and significant lethargy lasting approximately 30 h. A 6-year-old who received two doses of aripiprazole therapeutically experienced lethargy, drooling, and flaccid facial muscles which improved with diphenhydramine. Two adolescents remained asymptomatic despite doses of 120 mg and 300 mg while a third adolescent with an unknown dose experienced transient lethargy. CONCLUSION: Aripiprazole is capable of producing marked lethargy and gastrointestinal upset in pediatric patients. Adolescents in this series experienced only minor, if any, clinical effects. Major clinical effects, i.e., seizures, dysrhythmias, were not reported in this series. PMID- 15902788 TI - Delayed neurologic sequelae resulting from epidemic diethylene glycol poisoning. AB - BACKGROUND: Diethylene glycol (DEG) is a well-known metabolic and renal toxin usually ingested accidentally as an ethanol substitute or as a contaminant in various medicinals. To date, most poisonings have occurred in third-world countries where early death from renal failure is very common. We report a series of seven patients presenting with epidemic DEG poisoning from a correctional facility with varying degrees of metabolic acidemia and acute renal impairment responding to emergent hemodialysis (HD). Significantly, three patients developed delayed neurologic toxicity which has not been well characterized in the past. CASE SERIES: Seven male patients (age range 19-55) presented over a 36 h period following ingestion of varying quantities of DEG. Initially three patients, ingesting the largest quantities of DEG, presented more than 24 h postingestion with severe metabolic acidemia (pH range 6.8-7.1) and anuric acute renal failure requiring HD. All three remained dialysis-dependent and developed significant cranial neuropathies with bulbar palsy in the second week postingestion. One patient died with cerebral oedema and a progressive encephalopathy. Two further patients presented within 24 h of ingestion with normal renal function and a moderate metabolic acidemia (pH range 7.2-7.28) requiring HD. They remained well. Finally, two further patients presented with a history of trivial DEG ingestion and did not require any therapy. Neurologic signs in the two surviving initial presenters improved over 4-6 months although they remained dialysis-dependent. CONCLUSION: Unrecognized DEG poisoning may present with metabolic acidemia and anuric acute renal failure. Established renal impairment may predict subsequent delayed neurologic toxicity. PMID- 15902789 TI - Does ethanol explain the acidosis commonly seen in ethanol-intoxicated patients? AB - OBJECTIVE: Emergency physicians frequently treat ethanol-intoxicated trauma patients. In patients with apparently minor injuries, the presence of metabolic acidosis is often attributed to serum ethanol. We tested whether there is justification for the bias that ethanol reliably explains the acidosis commonly seen in alcohol-intoxicated patients. METHODS: Prospective, observational. INCLUSION CRITERIA: Ethanol-intoxicated patients admitted to the emergency department (ED) following significant trauma mechanisms, in whom diagnostic evaluation revealed only minor injury. EXCLUSION CRITERIA: Major trauma (blood transfusions, drop in Hct > 10 points over 24 h, or Injury Severity Score [ISS] >5) or positive urine toxicology screen. DEFINITIONS: Ethanol Intoxication: (Blood Alcohol Level (BAL) > or =80 mg/dl), Acidosis: BD < or = -3.0 mMol/L; Lactic Acidosis (LAC >2.2 mMol/L). Data were reported as mean+/-SD. Data were compared by t-tests or Fishers exact test as appropriate (alpha= 0.05, 2 tails) and correlations by Pearson correlation coefficient. RESULTS: 192 patients were studied (84% male) with a mean age of 31.7+/-15.6 years. Acidosis was observed in 19.3% (CI 95%, 14.5% to 25.0%) of all study patients. We observed significant (p<0.001) difference in prevalence of acidosis in ethanol intoxicated (42%) compared to nonintoxicated (1%) patients. Comparing the two study groups, patients with ethanol intoxication had lower BD (-2.24+/-2.74 vs. -0.05+/-2.35, p<0.001) and higher LAC (2.69+/-1.48 vs. 2.00+/-1.78, p=0.02). However, ethanol levels did not correlate significantly with BD (p=0.50) or LAC (p=0.14). CONCLUSION: Ethanol intoxication is associated with acidosis, which does not correlate with BD or LAC. The complexity of pathogenesis of acidosis in ethanol intoxication justifies further diagnostic evaluation of these patients in order to rule out other causes of acidosis. PMID- 15902790 TI - False positive acetaminophen levels associated with hyperbilirubinemia. AB - Serum acetaminophen determination is frequently necessary in patients with hepatic failure. We observed two patients (#1, #2) with elevated serum total bilirubin levels (26.5 mg/dL and 40.1 mg/dL) who had multiple false positive acetaminophen levels using the kinetic method of the GDS Diagnostics enzymatic acetaminophen assay (GDS Diagnostics, Elkhart, IN). We investigated the magnitude, threshold, and linearity of this effect using the GDS Diagnostics assay and an EMIT acetaminophen assay on two other hyperbilirubinemic patients (#3, #4) and a commercial solubilized bilirubin standard. Samples were diluted using fresh frozen plasma, and acetaminophen levels were analyzed twice using the kinetic method of the GDS Diagnostic acetaminophen assay and twice with the EMIT assay. The absence of acetaminophen in all samples was verified by gas chromatography/mass spectroscopy (GC/MS). The kinetic GDS assay resulted in a positive acetaminophen assay (cutoff for a positive result= 10 mg/L) with patient #3, patient #4, and in the bilirubin standard when the total bilirubin levels were 28.2 mg/dL, 22.5 mg/dL, and 18.3 mg/dL, respectively. One sample was interpolated to give a positive acetaminophen reading when diluted to a total bilirubin concentration of 15 mg/L. None of the samples tested with GC/MS or the EMIT assay resulted in any detectable acetaminophen. In conclusion, caution must be taken utilizing the GDS Diagnostic assay for the quantification of acetaminophen with concomitant hyperbilirubinemia. Alternatives such as EMIT or GC/MS should be employed to assess acetaminophen levels in such patients. PMID- 15902791 TI - A toxicologist's guide to studying diagnostic tests. AB - Diagnostic testing plays an important role in medicine, including in clinical toxicology. In comparison with therapeutics however, until recently, much less focus has been input in this vital area of research and practice. In the absence of stringent and constructive criteria, the quality and interpretability of studies evaluating diagnostic accuracy has been less than optimal. Recently, a group of researchers and clinicians have attempted to outline a checklist intended to provide guidelines for undertaking and reporting studies examining diagnostic accuracy. Also, a tool to assess the quality of diagnostic accuracy studies was developed. The purpose of this review is to inform practitioners and researchers in the field of clinical toxicology on the basic concepts underlying measurements of the accuracy of diagnostic tests, methodological concerns related to design and analysis of studies, the strengths and weaknesses of different measures of accuracy and an overview of the Standards for Reporting Diagnostic Accuracy (STARD statement) and the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) tool. Understanding key concepts in the field of diagnostic testing can not only help improve future studies but also explain why diagnostic tests currently used by clinical toxicologists are not performing well in particular circumstances. PMID- 15902792 TI - Hyperbaric oxygen in the treatment of carbon monoxide poisoning. AB - Carbon monoxide, a byproduct of incomplete hydrocarbon combustion, has been responsible for many accidental poisonings worldwide. The signs and symptoms of poisoning are diverse, ranging from headache, dizziness, and confusion to cardiac and neurological disturbances. Oxygen is the cornerstone of treatment, because it accelerates the dissociation of carbon monoxide from heme proteins. The role of hyperbaric oxygen in the treatment of CO poisoning is still questionable. Only a few randomized, controlled studies have been conducted, and their results are inconsistent. In the present review, we discuss the conclusions of four randomized controlled studies and propose a hyperbaric oxygen treatment protocol based on these conclusions. PMID- 15902793 TI - Acute felbamate overdose with crystalluria. AB - A 3-year-old child developed vomiting, ataxia, and crystalluria after ingestion of approximately 232 mg/kg of felbamate elixir. High-powered polarization microscopy of the urine revealed sharp, needle-like crystals. The analysis of the urine crystals showed unchanged felbamate (80.9%), monocarbamate felbamate (18.8%), and trace amounts of mercapturic acid conjugates of the metabolite 2 phenylpropenal (0.1%). The serum felbamate level 15 h after ingestion was 138 mg/L. Crystalluria and hematuria resolved with intravenous fluid therapy, and the child recovered within 24 h. PMID- 15902794 TI - Aripiprazole (abilify) overdose in a child. AB - Aripiprazole, 7-{4-[4-(2,3-dichlorophenyl)-1-piperazinyl] butyloxy}-3,4-dihydro 2(1H)-quinolinone, is an atypical antipsychotic medication inaugurating the newest class of atypical antipsychotics: the partial dopamine agonists. It has a particularly long half-life of elimination and variable metabolism secondary to genetic polymorphism. We report an unintentional overdose of 195 mg (17.1 mg/kg) of aripiprazole in a 2.5 year-old child. This patient exhibited CNS depression not requiring respiratory support and without significant cardiovascular effects. CNS effects persisted for almost 2 weeks postingestion. PMID- 15902795 TI - Chronic renal disease patients with severe star fruit poisoning: hemoperfusion may be an effective alternative therapy. AB - We report a case of star fruit intoxication in a 60-yr-old male patient with a past medical history of diabetes mellitus and chronic renal failure. Clinical effects included hiccups, hearing impairment, urine retention, and disturbed consciousness. Star fruit intoxication was also the suspected cause of hypothermia, an unusual symptom. The patient remained comatose after receiving two sessions of hemodialysis. However, after a 6 h session of charcoal hemoperfusion following the second hemodialysis treatment, his consciousness returned to normal within 1 day. While no previous study on hemoperfusion therapy in star fruit intoxication has been reported, in view of the fatal outcome of star fruit intoxication in uremic patients, hemoperfusion may be an alternative therapy if intensified hemodialysis fails. PMID- 15902796 TI - Conservative management of delayed, multicomponent coagulopathy following rattlesnake envenomation. AB - BACKGROUND: Crotaline Fab therapy is recommended for controlling local tissue effects, coagulation abnormalities, and other systemic signs following mild-to moderate N American Crotaline envenomations. Occasionally, coagulation abnormalities emerge after control of tissue effects has been achieved. These coagulation changes range from minor, single parameter abnormalities to multicomponent, critical value derangements. The bleeding risk associated with these abnormalities is unknown, and dosing guidelines for Crotaline Fab therapy in treating coagulation abnormalities that are severe or delayed-in-onset have not been clearly established. CASE REPORT: A 40-year-old man was envenomated in the right hand by a rattlesnake. Crotaline Fab therapy was started within 1 h of envenomation and arrest of edema progression was achieved by 48 h of admission. Although fibrinogen, platelet count, aPTT, and INR were all normal on initial assessment, hematologic changes were evolving over post-envenomation days 1 to 5 despite 32 vials of Crotaline Fab. The patient was transferred to our tertiary care center on the fifth postenvenomation day with a platelet count of 15,000/mm3 and unmeasurable values for fibrinogen, aPTT, and INR. The patient was managed with close observation alone, with no additional antivenom, and was discharged home on day 12 with no improvement in fibrinogen, aPTT or INR. Follow up lab assessment showed resolution of the hematologic effects sometime between day 17 and 37. CONCLUSION: We report a case of rattlesnake envenomation with profound, delayed hematologic effects that were resistant to 32 vials of Crotaline Fab given over post-envenomation days 1 to 4. After day 4, no further attempt was made to achieve normal lab indices using antivenom. Close observation alone may be adequate in cases of multicomponent, critical value coagulopathies following rattlesnake envenomation as long as there is no evidence of bleeding and local tissue effects and systemic effects have been adequately controlled. PMID- 15902797 TI - Myocardial ischemia as a result of severe benzodiazepine and opioid withdrawal. AB - Long-term infusion of benzodiazepines and opioids is strongly associated with dependence and withdrawal syndromes. We report the first case of severe benzodiazepine and opioid withdrawal resulting in transient myocardial ischemia. CASE REPORT: A 6-month-old female born at 25 weeks gestation with severe opioid and benzodiazepine dependence resulting from multiple operative procedures and chronic ventilatory support was receiving continuous intravenous infusion of fentanyl and midazolam after trials of enteral methadone and diazepam had been unsuccessful due to gastric intolerance. On postoperative day 5 following Nissen fundoplication and gastrostomy tube placement, she acutely developed tachycardia, hypertension, agitation, loose stools, and yawning. Attempts to provide boluses of benzodiazepines and opioids revealed a very sluggish port in her subclavian central venous catheter. Prompt replacement of the catheter occurred without complication. After resuming infusions and providing additional sedatives and opioids, the loose stools, yawning, and agitation resolved. However, the tachycardia persisted. A 12-lead ECG was notable for significant ST depression in anterior leads. Laboratory studies revealed significantly elevated cardiac enzymes. The patient was transfused with packed red blood cells to optimize oxygen-carrying capacity. Echocardiography demonstrated a small region of dyskinetic apical endocardium. Cardiac enzymes normalized within 48 h. The ECG and echocardiographic findings fully resolved after approximately 70 h. DISCUSSION: We believe that the sluggish central venous catheter port limited delivery of the midazolam and fentanyl to our patient. The resultant tachycardia and hypertension limited diastolic filling of the coronary arteries, resulting in myocardial ischemia. As the withdrawal was treated, heart rate and blood pressure returned to baseline, myocardial perfusion normalized, and the ST depression and the cardiac enzyme values normalized. This report underscores the significant morbidity associated with withdrawal syndromes and the need to recognize withdrawal early and to treat it aggressively. PMID- 15902798 TI - Millipede exposure. PMID- 15902799 TI - Comment on "refractory hypoglycemia from ciprofloxacin and glyburide interaction". PMID- 15902800 TI - FFP in organophosphate poisoning: what's the secret ingredient? PMID- 15902801 TI - Comment on "Acetaminophen: the 150 mg/kg myth". PMID- 15902802 TI - Poisoning with insulin glargine. PMID- 15902803 TI - Key advances in care for the elderly. PMID- 15902804 TI - The patient with intermittent claudication. PMID- 15902805 TI - Knowing when to treat ear wax. PMID- 15902806 TI - Reducing the risk pressure ulceration. PMID- 15902807 TI - Urinary incontinence in the female patient. PMID- 15902808 TI - Managing the erythrocytoses. PMID- 15902809 TI - Current perspectives in bipolar disorder. PMID- 15902810 TI - Managing summer hayfever. PMID- 15902811 TI - Life beyond the MRCGP. PMID- 15902813 TI - 2004: that was the year that was. PMID- 15902814 TI - A trainee's perspective on off-pump coronary artery bypass. PMID- 15902815 TI - Sirolimus-eluting stents for treatment of in-stent restenosis: immediate and late results. AB - We analyzed the clinical, angiographic, and late intravascular ultrasonographic findings from 140 patients whose in-stent restenosis was treated with sirolimus eluting stents. In-stent restenosis remains the main limitation to percutaneous coronary revascularization and has a high recurrence rate after bare stent implantation. From May 2002 through July 2003, we studied 140 patients with clinical restenosis after bare-stent treatment. In 107 patients, in-stent restenosis occurred de novo; in 28 patients, this was the 2nd restenosis; and in another 5, it was the 3rd occurrence. A sirolimus-eluting stent was implanted directly after angiographic evaluation of the in-stent restenosis in 79 patients and after pre-dilation in 61 patients. All patients were given the following antithrombotic regimen: low-molecular-weight heparin, ticlopidine, and aspirin for 1 month, followed by clopidogrel and aspirin for 1 year. Primary success was achieved in 137 patients. Three patients had a non-Q wave myocardial infarction. At the 1-month evaluation, 2 patients had died: 1 due to subacute stent thrombosis and another due to acute mesenteric ischemia. After a mean follow-up of 16 +/- 4 months, the major adverse cardiac events were acute myocardial infarction due to late stent thrombosis in 2 patients and the need for target lesion revascularization in 15 patients. Late angiographic evaluation was performed in 97 patients (69%), 16 of whom had new restenosis: 14 of the restenoses were intrastent, and 2 were at the edges of the stent. Our results suggest that sirolimus-eluting stents are effective in the prevention of in-stent restenosis and, therefore, may become the leading treatment alternative for patients with in-stent restenosis. PMID- 15902816 TI - Pediatric aortic disruption. AB - Although trauma is the primary cause of death in children, few reports or series exist regarding the management of traumatic aortic disruption in the pediatric age group. The clinical outcome in children diagnosed with acute aortic disruption may be directly influenced by diagnostic and therapeutic management decisions. We reviewed the clinical course of 3 consecutive pediatric patients (mean age, 10.0 years; range, 4-16 years) admitted to our institution from January 2002 through May 2003 with the diagnosis of acute aortic disruption due to blunt trauma. In each case, the cause was a motor vehicle accident. Major, concomitant injuries involving other organ systems were present in each patient. Our operative goals were to use primary repair techniques, avoid the use of endovascular stent grafts, and use partial left heart bypass during aortic cross clamping whenever possible. Each patient underwent successful operative repair. Aortic reconstruction techniques included primary suture repair in the 4-year-old patient, patch angioplasty in the 16-year-old, and placement of an interposition conduit in the 10-year-old for a blow-out type aortic injury. All patients received partial left heart bypass during aortic cross-clamping (mean, 36.6 min; range, 27-50 min), via a centrifugal pump, and anticoagulation. All patients recovered without evidence of adverse neurologic sequelae. Operative repair of acute aortic disruption in pediatric patients using circulatory support can provide good outcomes. Although not always feasible, the preferential use of primary aortic repair techniques in lieu of interposition conduits and endovascular aortic stents may decrease the potential for late pseudocoarctation. PMID- 15902817 TI - Ectasia and severe atherosclerosis: relationships with chlamydia pneumoniae, helicobacterpylori, and inflammatory markers. AB - To date, there has been no convincing evidence for an association between Chlamydia pneumoniae or Helicobacter pylori and ectasia. In this case-control study, we have investigated the association of H. pylori and C. pneumoniae seropositivity with ectasia, severe coronary atherosclerosis, and normal vessels, which were so classified by coronary angiography. We have also evaluated the influence of these infections on inflammatory markers such as high-sensitive C reactive protein (hsCRP) and interleukin 6 (IL-6). Of the 796 patients undergoing coronary angiography for suspected ischemic heart disease, 244 patients were recruited. Of these, 91 had normal vessels, 88 had 3 or more obstructed vessels, and 65 had ectatic vessels without atherosclerosis. Eighty-seven atherosclerotic patients (98.9%) were positive for C. pneumoniae IgG, as were 64 ectatic patients (98.5%) and 76 controls (83.5%) (P < 0.001). Forty-two atherosclerotic patients (47.7%) were positive for C. pneumoniae IgM, as were 43 ectatic patients (66.2%) and 43 controls (47.3%) (P = 0.036). Seventy-two atherosclerotic patients (81.8%) were positive for H. pylori IgA, as were 26 ectatic patients (40.0%) and 44 controls (48.4%) (P < 0.001). High-sensitive CRP levels were significantly higher in ectatic patients (5.639 mg/L) than in controls (4.390 mg/L) (P = 0.032), and IL-6 levels were significantly higher in atherosclerotic patients (33.92 U/L) than in controls (14.01 U/L) (P < 0.001). Interleukin-6 levels were higher in H. pylori seropositive patients, and hsCRP levels were higher in C. pneumoniae seropositive patients, when compared with seronegatives. We suggest that, as in atherosclerosis, C. pneumoniae infection is related to ectasia, with raised CRP levels. PMID- 15902818 TI - Left ventricular diastolic function and circadian variation of blood pressure in essential hypertension. AB - The purpose of this prospective study was to determine the relationship between circadian blood pressure and left ventricular diastolic function in essential hypertension. The study population included 25 patients aged 56 +/- 18 years with non-dipper hypertension and 25 age- and sex-matched patients with dipper hypertension. They underwent conventional Doppler echocardiography and color tissue Doppler from apical 4- and 2-chamber views. In non-dipper patients, diastolic left ventricular function was reduced significantly. The transmitral E wave decreased (0.55 +/- 0.2 vs 0.62 +/- 0.2 m/s, P < 0.05), the transmitral A wave increased (0.77 +/- 0.1 vs 0.70 +/- 0.1 m/s, P < 0.01), the transmitral E/A ratio decreased (0.78 +/- 0.1 vs 0.86 +/- 0.2, P < 0.05), and the transmitral E wave deceleration time increased in non-dipper patients (211 +/- 44 vs 196 +/- 42 ms, P < 0.05). The isovolumic relaxation time increased (112 +/- 15 vs 105 +/- 14 m/s, P < 0.05). The mean left ventricular myocardial velocities also differed significantly; the early diastolic velocity decreased (5.9 +/- 2. 1 vs 77 +/- 3.1 cm/s, P < 0.01), the late diastolic velocity increased (9.5 +/- 2.7 vs 8.7 +/- 1.6 cm/s, P < 0.05), and the E/A ratio decreased (0.68 +/- 0.55 vs 0.94 +/- 0.39, P < 0.01). These findings suggest that non-dipper hypertensive patients who have impaired left ventricular diastolic function should be identified early for careful follow-up and possible referral to a specialized center. PMID- 15902819 TI - Effects of previously well-developed collateral vessels on left internal mammary artery graft flow after bypass surgery. AB - Transthoracic Doppler ultrasonography can assess left internal mammary artery patency and flow after coronary artery bypass grafting. We aimed to show, by transthoracic Doppler ultrasonography, the early effects upon left internal mammary artery graft flow of preoperative collateral vessels supplying the left anterior descending artery. Thirty-four consecutive patients undergoing coronary artery bypass were prospectively enrolled: 19 patients with collateral vessels supplying the left anterior descending were compared with 15 patients without collaterals. After bypass, end-diastolic velocity, mean velocity, flow volume, and ejection fraction were significantly greater, and the resistivity index was lower in patients with collateral vessels. The changes in velocities, volume, resistivity index, and pulsatility index were also found to be greater in patients with collateral vessels than in those without collaterals. Collateral vessels were the only factor affecting the changes in end-diastolic volume, mean velocity, flow volume, and resistivity index in multivariate analysis. Three factors affected postoperative left ventricular ejection fraction: collateral vessels, preoperative ejection fraction, and changes in left internal mammary flow volume. We conclude that patients with well-developed collaterals to the left anterior descending have better flow in the left internal mammary graft and more significant improvement in left ventricular function after coronary bypass. The flow volume of the mammary graft and the improvement of ventricular systolic functions after coronary bypass might be presumed with the presence of grade 2 or 3 preoperative collateral vessels. PMID- 15902820 TI - Double-patch repair of postinfarction ventricular septal defect. AB - We report 4 consecutive cases in which the double-patch technique was used to repair an inferior postinfarction ventricular septal rupture. The ventricular septal perforation was closed directly by stitching, with the same sutures, 2 autologous pericardial patches onto both sides of the affected septum, through only a left ventriculotomy. Complete closure of the defect was accomplished, and no residual shunt was observed in any patient. This technique appears to be useful in selected cases, such as ventricular septal perforation with myocardial infarction in the subacute or chronic phase, especially in instances of inferoposterior infarction. Further experience is needed to verify its safety and efficacy. PMID- 15902821 TI - Systolic anterior motion of the mitral valve after mitral valve repair: a method of prevention. AB - Factors predisposing patients to systolic anterior motion of the mitral valve (SAM) with left ventricular outflow tract (LVOT) obstruction after mitral valve repair are the presence of a myxomatous mitral valve with redundant leaflets, a nondilated hyperdynamic left ventricle, and a short distance between the mitral valve coaptation point and the ventricular septum after repair. From December 1999 through March 2000, we used our surgical method in 6 patients with severely myxomatous regurgitant mitral valves who were at risk of developing SAM. Leaflets were markedly redundant in all 6. Left ventricular function was hyperdynamic in 4 patients and normal in 2. Triangular or quadrangular resection of the midportion of the posterior leaflet and posterior band annuloplasty were performed. To prevent SAM and LVOT obstruction, extra, posteriorly directed, mid-posterior leaflet secondary chordae tendineae, which would otherwise have been resected, were transferred to the underside of the middle of the mid-anterior leaflet with a small piece of associated valve as an anchoring pledget. This kept the redundant anterior leaflet edge, which extended below the coaptation point, away from the LVOT No post-repair SAM or LVOT obstruction was observed on intraoperative or discharge echocardiography. All patients had no or trivial residual mitral regurgitation. We conclude that extra chordae tendineae, when available, can be used in mitral valve repair to tether the redundant anterior leaflet and thus prevent it from flipping into the LVOT. This will theoretically prevent SAM and LVOT obstruction in patients with risk factors for SAM. PMID- 15902822 TI - Increased myocardial ischemia during nitrate therapy: caused by multiple coronary artery-left ventricle fistulae? AB - We present the case of a 54-year-old man who had crescendo angina during nitrate therapy. Selective coronary angiograms showed no atherosclerotic lesions, but did show plexuses of intramural vessels that connected the distal thirds of the left and right coronary systems with the left ventricle. The cause of our patient's increased myocardial ischemia during nitrate therapy may have been the coronary "steal" phenomenon. PMID- 15902823 TI - Questions on coronary fistulae and microfistulae. PMID- 15902824 TI - Congenital circumflex coronary arteriovenous fistula with aneurysmal termination in the pulmonary artery. AB - Coronary arteriovenous fistula is a rare congenital anomaly that is seen in 0.1 % to 0.2% of coronary angiograms. Aneurysmal formation in the fistula is even rarer. We report a case of congenital circumflex arteriovenous fistula with aneurysmal formation just near its termination in the pulmonary artery, associated with atherosclerotic left anterior descending coronary artery. The anomaly was successfully repaired. PMID- 15902825 TI - Retrospective analysis of 286 patients requiring circulatory support with the intraaortic balloon pump. 1977. PMID- 15902826 TI - Abdominal Aortic dissections: anatomic and clinical features and therapeutic options. AB - Isolated abdominal aortic dissections are rare events. Their anatomic and clinical features are different from those of atherosclerotic aneurysms. We report 4 cases of isolated abdominal aortic dissection that were successfully treated with surgical or endovascular intervention. The anatomic and clinical features and a review of the literature are also presented. PMID- 15902827 TI - Coronary thrombosis related to use of Xenadrine RFA. AB - Recently, ephedra was removed from the U.S. marketplace due to a heightened concern that dietary supplements containing ephedra may present "an unreasonable risk of illness or injury." This is the 1st time the U.S. Food and Drug Administration has banned an herbal supplement, and the ban sheds light on the potential harm of nutritional supplements that are used for weight loss or as a boost to athletic performance. We report the case of a body builder who used Xenadrine RFA, an ephedra-containing supplement, at recommended doses for nearly a year; he then experienced an acute myocardial infarction, which was documented to be secondary to thrombosis in situ. We ruled out other possible causes of myocardial infarction, as well a hypercoagulable state. There was no evidence of illicit drug use. Our report serves as a poignant reminder of the potential dangers of herbal supplementation, especially when used to heighten athletic performance. PMID- 15902828 TI - Cardiogenic shock caused by severe coronary artery spasm immediately after coronary stenting. AB - Coronary artery spasm is common during percutaneous coronary intervention and is easily relieved by intracoronary administration of vasodilators. We report the case of a patient who had severe, protracted, generalized spasm of the entire left coronary artery system during coronary artery stenting. The spasm, which was unresponsive to intracoronary vasodilators administered via guiding catheter, resulted in pulmonary edema and cardiogenic shock. Local injection of nitroglycerin via a transit catheter in the coronary artery eventually resolved the spasm and reversed the cardiogenic shock. To our knowledge, this is the 1st report of such a case in the English-language medical literature. PMID- 15902829 TI - Penetrating aortic ulcers after absorption of intramural hematoma. AB - We report a case of a type B aortic intramural hematoma, which rapidly expanded, with ulcer-like projections, after complete absorption of the hematoma. One month after the initial presentation, a new computed tomographic scan showed the appearance of a thin ulcer-like projection in the aortic wall. Three months later, after the patient reported a new episode of chest pain, a computed tomographic scan revealed 2 penetrating ulcers and rapid aortic dilation; the aortic hematoma had been completely absorbed. Urgent thoracic aortic replacement was undertaken. Three years postoperatively, the patient was asymptomatic, with no lesion or dilation of the aorta upon computed tomography. After an acute hematoma, the strength and structure of the aortic wall can alter rapidly. Damage and weakening of the aortic wall are caused mainly by infiltration of inflammatory cells, which have pronounced proteolytic and elastolytic activity. Due to the unstable nature of the lesion, optimal management remains controversial. In our patient, the inflammatory process led to the development of 2 aortic ulcers and aortic dilation within 3 months of the acute lesion, requiring urgent surgical intervention. PMID- 15902830 TI - Adrenal adenoma presenting with ventricular fibrillation. AB - We report the case of a 58-year-old man who presented with ventricular fibrillation. The serum potassium level was 1.8 mEq/L after successful cardioversion. Coronary angiography showed a normal heart with no structural defects, but 12-lead electrocardiography showed indications of left ventricular hypertrophy, which was confirmed by echocardiography. Laboratory examinations showed a suppressed renin level and an elevated serum aldosterone level. Computed tomography then revealed a right adrenal mass. The patient was treated with surgical resection of the adenoma. This case emphasizes the importance of meticulous search for secondary causes of hypertension, before the occurrence of serious complications. PMID- 15902831 TI - Transpulmonary surgical closure of patent ductus arteriosus with hypothermic circulatory arrest in an adult patient. AB - Untreated patent ductus arteriosus carries a higher risk in adults than in children, especially when the defect is large (>4.0 mm in diameter), short, or friable. Therefore, various technical precautions have been suggested for application during surgical closure of a patent ductus arteriosus in an adult. We report the case of a 47-year-old woman with a patent ductus arteriosus who underwent transpulmonary surgical closure of the ductus under hypothermic total circulatory arrest. We discuss the technique in light of the current English language medical literature. PMID- 15902832 TI - Ultrafast computed tomography in management of post-bentall aortic root pseudoaneurysm repair. AB - A giant pseudoaneurysm adjacent to the sternum was diagnosed in a patient who had undergone aortic root replacement with use of the Bentall operation 10 years earlier. Electron-beam computed tomography showed that the right coronary artery, which could not be seen on angiography, originated from the pseudoaneurysm itself. At reoperation, we found that both coronary ostia were detached and that the right coronary ostium was totally detached. The source of the right coronary artery blood flow was the pseudoaneurysm itself The defect at the left coronary artery attachment site was repaired primarily. A vein graft was interposed between the ostium of the right coronary artery and the native aorta, distal to the graft anastomosis. PMID- 15902833 TI - Large pseudoaneurysm of the carotid artery in Behcet's disease. AB - Behcet's disease is a multisystem inflammatory disease of unknown cause. Its major pathologic feature is vasculitis. We report the occurrence of a large pseudoaneurysm of the carotid artery in a patient who had Behcet's disease. The patient underwent surgery and was discharged on the 7th postoperative day. Ours is only the 4th such case reported in the English-language medical literature. Endovascular and surgical options are discussed herein. PMID- 15902834 TI - Thirty-seven-year durability of a Starr-Edwards aortic prosthesis: case report and brief review of the literature. AB - We report the case of a patient who was diagnosed with prosthetic valve endocarditis caused by Staphylococcus aureus. He had undergone aortic valve replacement with a Starr-Edwards prosthesis 37 years earlier Because of uncontrolled infection despite antibiotic treatment, the patient underwent successful surgical replacement of the prosthetic valve. PMID- 15902835 TI - Transcatheter palliation of tetralogy of Fallot with pulmonary artery discontinuity. PMID- 15902836 TI - Prominent J wave (Osborn wave) with coincidental hypothermia in a 64-year-old woman. PMID- 15902837 TI - Chest pain and diaphragmatic pacing after pacemaker implantation. PMID- 15902838 TI - A potential complication of directional coronary atherectomy for in-stent restenosis. PMID- 15902839 TI - Aortic root infection destroying the left fibrous trigone. PMID- 15902840 TI - Recurrent primary vascular echinococcosis. PMID- 15902841 TI - Tricuspid repair and cavopulmonary anastomosis after blunt trauma. PMID- 15902842 TI - Role of endothelin in fibrosis and anti-fibrotic potential of bosentan. AB - Recent data demonstrate the fundamental role of endothelin in the pathogenesis of fibrosis, and the anti-fibrotic potential of dual endothelin receptor antagonists such as bosentan. Although transforming growth factor-beta, aldosterone and connective tissue growth factor, have already been established as contributors to the process of fibrosis, endothelin now emerges as a key player, which may have a role both in the initiation and in maintenance of fibrosis, and may mediate the pro-fibrotic effects of the other agents. Bosentan is an orally active, dual endothelin receptor antagonist, which competitively antagonizes the binding of endothelin to both endothelin receptors ETA and ETB. Bosentan prevents endothelin induced fibroblast proliferation and extracellular matrix deposition and contraction, and reduces cardiac, hepatic, pulmonary and renal fibrosis in different disease models characterized by the activation of the endothelin system. Bosentan even reverses existing fibrosis, possibly by its effect of stimulating matrix metalloproteinase type 1 (collagenase) expression. The anti fibrotic effects of bosentan extend to fibrosis induced by mediators other than endothelin such as transforming growth factor-beta, angiotensin II and aldosterone, indicating a central role of endothelin and endothelin receptors in fibrotic processes. PMID- 15902843 TI - Acute renal failure in cancer patients. AB - Acute renal failure (ARF) is defined as a sudden decrease in glomerular filtration rate leading to an acute rise in blood urea nitrogen and serum creatinine levels. It is a serious complication of cancer and constitutes a major source of morbidity and mortality. Current data suggest that ARF has the potential to substantially jeopardize the chances of cancer patients receiving optimal treatment and a potential cure. The pathways leading to ARF in cancer patients are common to the development of ARF in other conditions. However, ARF may also develop due to aetiologies arising from cancer treatment or the disease itself, including: nephrotoxic chemotherapy agents, post-renal obstruction, compression and infiltration by malignancy, tumour lysis syndrome, uric acid, sepsis and contrast agent nephropathy. This review provides a comprehensive overview of the causes of ARF in patients with cancer and guidance on how to prevent and treat this condition. Ultimately, the key to managing ARF in cancer patients is to ensure that a multidisciplinary approach provides adequate assessment, appropriate preventative measures and early intervention. PMID- 15902844 TI - On roots of childhood asthma: the role of respiratory infections. AB - The prevalence of childhood asthma has increased throughout the last decades, but the reasons for the increase still remain unresolved. However, the debate has recently focused on the role of infections, and microbial exposure in general, in the development of childhood asthma. Many studies have suggested that there is an inverse relation between certain infections and the risk of asthma. This article outlines the contemporary role of childhood asthma in public health, reviews the importance of topical respiratory pathogens in the development of childhood asthma, and provides an example how selection bias may have caused controversy in previous studies assessing the role of infections in the development of childhood asthma. PMID- 15902845 TI - Adenoviruses for treatment of cancer. AB - Most cases of cancer, when detected at an advanced stage, cannot be cured with conventional therapeutic modalities. Therefore, novel targeted approaches such as gene therapy are needed. Nevertheless, while the safety record of gene therapy for cancer has been excellent with more than a thousand patients treated without mortality related to the therapy, clinical efficacy has so far been limited. Moreover, it has become evident that clinical efficacy is partly determined by efficacy of gene delivery. Most adenoviruses used for gene therapy have been based on serotype 5 (Ad5). Unfortunately, recent data suggest that the primary receptor, the coxsackie-adenovirus receptor (CAR) expression in tumors may be highly variable resulting in resistance to adenovirus infection. Consequently, various strategies have been evaluated to modify adenovirus tropism in order to circumvent CAR deficiency, including retargeting complexes or genetic capsid modifications. To further improve tumor penetration and local amplification of the anti-tumor effect, selectively oncolytic agents, e.g. conditionally replicating adenoviruses (CRAds), have been constructed. Infection of tumor cells results in replication, oncolysis, and subsequent release of the virus progeny. Normal tissue is spared due to lack of replication. This review will focus on a discussion of various modifications of adenovirus to achieve efficient anti-tumor effect, and special emphasis will be placed on CRAds in multimodality treatments. PMID- 15902846 TI - Ion channels as novel therapeutic targets in heart failure. AB - Electrophysiological remodeling in heart failure (HF) is characterized by major changes in ion channel function and expression that alter the electrical phenotype and predispose to the development of lethal ventricular tachyarrhythmias. In this article, we provide a review of our current understanding of HF-induced ion channel dysfunction by highlighting changes in potassium and sodium currents, pumps, and exchangers as well as calcium handling proteins. We further relate these changes in ion channel function to abnormalities in impulse generation, conduction, and repolarization with a view towards identifying potentially novel targets for anti-arrhythmic therapy for this public health epidemic. PMID- 15902847 TI - Platelet adhesion in hypertension: application of a novel assay of platelet adhesion. AB - BACKGROUND: The increased risk of thromboembolism in hypertension may be related to a prothrombotic or hypercoagulable state, with abnormalities in haemostasis and platelet function. OBJECTIVE: To investigate the role of platelets in the pathogenesis of thrombosis in hypertension, we applied a novel new assay to detect and quantify the degree of platelet adhesion to a defined coagulation molecule. PATIENTS AND METHODS: Platelet-rich plasma (PRP) and citrated plasma (CP) were obtained from 50 patients with hypertension (25 treated, and 25 untreated) and 30 healthy controls. A suspension of 2 x 10(7) platelets were incubated for one hour in microtitre plates pre-coated with 5mg/mL fibrinogen. The supernatant was carefully aspirated, lysed with 5% tween and stored at -70 degrees C as supernatant platelet lysate (SPL). The wells were carefully washed with saline and bound platelets lysed as before, and stored at -70 degrees C as bound-platelet lysate (BPL). Soluble P-selectin (sP-sel) was determined in CP, SPL and BPL by enzyme-linked immunosorbent assay (ELISA). RESULTS: Patients with hypertension (both treated and previously untreated) had increased platelet adhesion, as determined by increased lysate sP-sel (P=0.002) in BPL, with no change in SPL (P=0.5) compared to healthy controls. There was no significant difference between treated and previously untreated hypertensives. CONCLUSION: Platelets from patients with hypertension display increased adhesion to an important coagulation factor (fibrinogen). This may, in part, account for the increased risk of thrombosis seen in these patients. PMID- 15902848 TI - Platelet microparticles and soluble P selectin in peripheral artery disease: relationship to extent of disease and platelet activation markers. AB - BACKGROUND: There is increased platelet activation in many cardiovascular diseases. This observation may explain the presence of increased levels of platelet microparticles (PMP) in these diseases. However, whether or not levels of PMPs inter-relate with other markers of platelet activation, such as soluble P selectin, or with disease severity, is unknown. We therefore hypothesized raised PMP levels in stable peripheral artery disease (PAD) intermittent claudication (IC), with an additional increase in severe PAD critical limb ischaemia (CLI). Furthermore, we tested the hypothesis that PMP levels are correlated with other markers of platelet activation, such as soluble P-selectin, membrane bound P selectin (CD62P) and 63. METHODS: Patients with PAD were recruited from the vascular outpatient and inpatient facilities at a teaching hospital. Age- and sex matched controls were also recruited from healthy volunteers. Venous blood was obtained from 23 patients with severe disease (CLI), 36 with moderate disease (IC), and from 30 healthy controls. The percentage of platelets positive for CD62P and CD63, as well as the numbers of PMPs were defined by flow cytometry. Plasma soluble P selectin was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: PMPs were increased relative to healthy controls in patients with IC, with a further increase in CLI (P<0.001). Soluble P selectin and CD62+ve platelets were raised in both patient groups, but there was no difference amongst the two patient groups. CD63+ve cells were raised only in CLI compared to healthy controls. In multivariate analysis, only PMP and soluble P selectin independently predicted disease severity, and the two markers correlated modestly (r=0.345, P<0.001). CONCLUSION: Increased PMP and soluble P selectin are both related to the severity of symptomatic PAD. However, it is uncertain if this relationship is a cause or effect of atherosclerosis. This finding may have clinical implications as PMPs have the potential to influence the progression of atheroma as well as promote thrombosis. PMID- 15902849 TI - A six-fold gradient in the incidence of type 1 diabetes at the eastern border of Finland. AB - OBJECTIVE: Type 1 diabetes results from gene-environment interactions in subjects with genetic susceptibility to the disease. We assessed the contribution of environmental and genetic factors to type 1 diabetes by comparing the incidence in two neighboring populations living in conspicuously different socioeconomic circumstances. RESEARCH DESIGN AND METHODS: We compared the incidence over a 10 year period (1990-99) in children younger than 15 years of age living in the Karelian Republic of Russia and in Finland. The frequency of susceptible and protective human leukocyte antigen (HLA)-DQ alleles was analyzed in 400 non diabetic schoolchildren from Russian Karelia and 1000 Finnish subjects. RESULTS: The average annual age-adjusted incidence of type 1 diabetes was lower in Russian Karelia than in Finland: 7.4 per 100000 (95% confidence interval 3.5-11.3) versus 41.4 per 100000 (37.3-45.5), while there were no differences in the frequency of the HLA DQ genotypes predisposing to type 1 diabetes in the background populations. The incidence rate did not differ significantly between different ethnic groups in Russian Karelia (Finns/Karelians, Russians, others). CONCLUSIONS: There is a close to six-fold gradient in the incidence of type 1 diabetes between Russian Karelia and Finland, although the predisposing HLA DQ genotypes are equally frequent in the two populations. This suggests that environmental factors contribute to this steep difference in the incidence rate between these adjacent regions. PMID- 15902850 TI - Functional results after revision of well-fixed components for stiffness after primary total knee arthroplasty. AB - Between 1990 and 2001, 16 well-fixed, aseptic, primary total knee arthroplasties were revised in 15 patients for a diagnosis of stiffness. Patients were followed for a mean of 42 months (range, 2-6 years). Of 15 patients, 10 (66%) were satisfied with the results of the procedure. The mean Knee Society pain score improved from 28 to 65 points, and the mean functional score improved from 45 to 58 points. The mean arc of motion improved from 40 degrees preoperatively to 73 degrees postoperatively. Recurrent stiffness required additional intervention in 4 knees (3 patients, 25%). The results of revision of a well-fixed, stiff, primary total knee arthroplasty were mixed in our hands and provided only modest improvements in pain, function, and arc of motion. Key words: knee, arthroplasty, stiffness, revision, arthrofibrosis. PMID- 15902851 TI - Minimally invasive total hip arthroplasty: a prospective randomized study. AB - Patients without prior hip surgery and body mass index lower than 30 undergoing primary total hip arthroplasty were eligible to participate in a randomized prospective study comparing a minimally invasive with a standard approach. The patients were randomized to receive incisions of 8 cm (group A, n = 28) or 15 cm (group B, n = 32). The groups were similar demographically. Patients in group A had significantly less intraoperative blood loss (P < .003) and less total blood loss (P < .009). Fewer patients in group A limped at 6 weeks (P < .04). Operative time, transfusion requirements, narcotic usage, length of hospital stay, achievement of rehabilitation milestones, cane usage, and complications were similar in both groups. There was no difference between the groups at 1- and 2 year follow-up. Compared with a standard incision, patients who underwent a minimally invasive total hip arthroplasty demonstrated decreased blood loss and limped less at 6-week follow-up. PMID- 15902852 TI - Comparison of mobile-bearing and fixed-bearing total knee arthroplasty: a prospective randomized study. AB - The purpose of this prospective randomized study was to compare the postoperative recovery and early results of 2 groups of patients undergoing total knee arthroplasty: 107 patients received an established fixed-bearing posterior stabilized prosthesis (Legacy Posterior Stabilized [LPS]), and 103 patients the meniscal-bearing prosthesis (Meniscal Bearing Knee [MBK]). Surgical procedures were the same for both groups except for posterior cruciate ligament management, which was sacrificed in the LPS group and spared but completely released from the tibia in the MBK group. At an average follow-up of 36 months, knee, function, and patellar scores were comparable in both groups. The LPS group showed a significantly higher maximum flexion than the MBK group (112 degrees vs 108 degrees). Using a fixed-bearing or a mobile-bearing design did not seem to influence the short-term recovery and early results after knee arthroplasty. Key words: total knee arthroplasty, mobile bearing, knee prosthesis, meniscal-bearing knee, posterior stabilized, prospective randomized. PMID- 15902853 TI - In vivo determination of knee kinematics for Japanese subjects having either a low contact stress rotating platform or an anteroposterior glide total knee arthroplasty. AB - The objective of this study was to work with a consecutive series of patients having Hospital for Special Surgery scores higher than 90 to evaluate kinematic patterns, under in vivo conditions, for 20 Japanese subjects implanted with 2 different mobile-bearing (MB) total knee arthroplasties (TKAs). Femorotibial contact paths for the medial and lateral condyles were then determined using a computer-automated model-fitting technique. This present study has shown that kinematic patterns for subjects having 2 different MB TKA designs differed but were not statistically different. Subjects implanted with a rotating platform (RP) MB TKA experienced minimal anteroposterior (AP) motion and larger axial rotation (RP). Subjects implanted with an anterior glide MB TKA experienced both femoral rotation and femoral translation (AP glide). There was minimal variability in the kinematic patterns for subjects implanted with an RP, whereas subjects implanted with an AP glide experienced more variable kinematic patterns. Key words: total knee arthroplasty, in vivo, fluoroscopy, kinematics. PMID- 15902854 TI - A 15- to 25- year follow-up study of primary Charnley low-friction arthroplasty: a single surgeon series. AB - Long term outcome studies are essential to determine the effectiveness and durability of a procedure. We report our 15 to 25 year clinical and radiographic follow-up with the Charnley low-friction hip arthroplasty. Four hundred and forty seven primary Charnley hip arthroplasties were performed by a single surgeon. Ninety-eight of the 447 hips from 75 patients were available for follow up at average of 18.9 years (range 15.3 to 25.4 years). Seventy-two hips (73.5%) survived clinically. Of these 72, seventy-one stems (98.6%) and 60 sockets (83.3%) survived radiographically. Kaplan-Meier survival analysis on all 447 hips using revision as an endpoint revealed 66.2% +/- 5.7% survival at 20 years. Twenty-six hips were revised. The main reason for revision was failure of the socket. Complications included dislocation (3 of 98 hips), heterotopic ossification (7 of 98 hips), trochanteric wire breakage (6 of 98 hips), and trochanteric non-union (1 of 98 hips). Charnley low-friction hip arthroplasty is an effectiveand durable procedure. Key words: hip, arthroplasty, charnley, follow up. PMID- 15902855 TI - The relationship between shelf life and in vivo wear for polyethylene acetabular liners. AB - Abstract: Although laboratory studies have correlated shelf life for gamma irradiated-in-air polyethylene with material degradation, it is unknown whether this clinically affects wear. Therefore, we examined the relationship between shelf life and clinical wear for 152 gamma-irradiated-in-air acetabular liners. True wear rates were calculated from computer-assisted head penetration analyses of serial radiographs. Follow-up averaged 6.6 years. Shelf life averaged 11.8 months with 96% of liners having shelf lives of less than 3 years. Data analysis revealed no correlation between shelf life and true wear rates (r2 = 0.002; P = .60). This study provides useful information for surgeons concerned about shelf life as a potential cause of high wear in previously implanted components. For conventional liners in inventory less than 3 years, we found no evidence of increased wear in vivo. PMID- 15902856 TI - Metal-on-Metal total hip replacement: what does the literature say? AB - Second-generation metal-on-metal (M/M) total hip replacements were introduced into clinical use in the late 1980s and demonstrate equivalent survivorship to conventional metal-on-polyethylene prostheses. Wear rates are comparable to those of first-generation designs that survived for a long time in the body. Biological effects from metal ions remain a concern. Patients with both first- and second generation M/M hips have higher levels of cobalt and chromium in their blood and urine than either patients with metal-on-polyethylene devices or unoperated patients. Concerns include the potential for acquired hypersensitivity, mutagenicity, and carcinogenicity. However, reports of proven adverse effects are scant. Prospective, randomized trials with follow-up in excess of 15 years will be needed to differentiate between the performance and effects of M/M and other bearing combinations. Key words: totalhip arthroplasty, metal-on-metal, wear, biological effects, clinical results. PMID- 15902857 TI - The role of intraoperative frozen section in decision making in revision hip and knee arthroplasties in a local community hospital. AB - There is little information in the literature regarding the reliability of intraoperative frozen section to predict infection in revision arthroplasties performed in community hospitals as most reports are from specialized centers or academic institutes. Between November 1997 and May 2001, we performed intraoperative frozen sections in 40 revision hip and knee arthroplasties. We used Mirra's criteria of more than 5 polymorphs per high power field to constitute a positive result. We found a sensitivity of 67% and a specificity of 97%. The positive and negative predictive values were 86% and 91%, respectively, with an accuracy of 90%. We conclude that intraoperative frozen section is a reasonably sensitive and specific adjunct investigation in the differentiation between septic and aseptic loosening. PMID- 15902858 TI - Application of core biopsy in revision arthroplasty for deep infection. AB - A preliminary core biopsy was performed in 168 consecutive joint arthroplasties (141 hips and 27 knees) before revision for suspected deep infection. Data of the core biopsy and subsequent revision procedures were available for 159 cases. Biopsies and revisions were all performed by one surgeon, and the exchange procedures were all undertaken in an ultra clean air enclosure. Biopsy material was cultured both aerobically and anaerobically and the results compared to that of the specimens obtained at the subsequent exchange procedure. In 116 patients (70%), the culture and sensitivity results were identical in samples from core biopsy and samples obtained during revision procedure (either no growth or growth of same organisms). When compared to intraoperative specimen report, 25 biopsies (15%) gave an apparently false-positive result. In 16 instances, a false-negative result was seen, but repeat biopsy in 4 of these cases was positive. The final diagnosis of infection was made using all available clinical and investigative data. When compared to final diagnosis, the sensitivity of core biopsy was 88%, the specificity was 91%, and the accuracy was 89%. Core biopsy was successful in identifying the organisms with the sensitivities to antibiotics in 80% of the disease-positive cases, hence, offering the opportunity to add the most appropriate antibiotic to bone cement during the exchange procedure. PMID- 15902859 TI - Polyethylene failure of the patellar component in New Jersey low-contact stress total knee arthroplasties. AB - In a postoperative 5- to 12-year follow-up study of 598 New Jersey low-contact stress total knee arthroplasties, 32 required revision. All retrieved patellar components were examined for polyethylene damage. These 32 cases had 3 types of failure: split rupture (7 cases), peripheral wear (21), and cantilevering breakage (4), respectively 75%, 64.3%, and 100% of which showed subluxation and/or tilting of the patellar component on the prerevision roentgenograms. Misalignment at the joint contact surfaces and rotational blockage of the mobile patellar component were considered the major causes of the failure. A design of a flatter metallic button (giving larger focal thickness of the polyethylene) and a dome-shaped polyethylene (reducing stress concentration at the pinnacle) may alleviate the failure driving mechanism should subluxation or tilting of the patellar component take place. PMID- 15902860 TI - Outcome of acetabular revision using an uncemented hydroxyapatite-coated component: two- to five-year results and review. AB - This is a retrospective review of 50 uncemented acetabular revisions with porous, hydroxyapatite-coated cups. The acetabulum alone was revised in 22 hips and both components were revised in 28 hips. The majority of hips (26) had type 2A (Paprosky) acetabular bone loss. All the revisions were carried out through a posterior approach. The mean duration of follow-up was 32 months (range, 24-52 months). Clinically, there was significant improvement in pain and moderate improvement in movement and mobility, and there is no radiological sign of failure of any cup so far. Re-revision was performed in 3 hips (6%) for recurrent dislocation. Our early results favor the use of this type of cup in acetabular revisions with moderate bone loss, but longer-term follow-up will be required. Key words: acetabulum, hydroxyapatite, revision, bone deficiency, complications. PMID- 15902861 TI - Periprosthetic bone remodeling around a prosthesis for distal femoral tumors: longitudinal follow-up. AB - Serial measurements using dual-energy x-ray absorptiometry (DEXA) were undertaken to evaluate progressive periprosthetic bone loss in patients treated for primary bone tumors of the distal femur using an uncemented tumor prosthesis. Twelve patients underwent sequential DEXA analysis on average 26.5 and 90.9 months postsurgery. Changes in bone mineral density were measured in regions of interest (ROIs) around the prosthesis stem. The test-retest reliability coefficient (r) ranged from 0.92 to 0.99 for all ROI. In the most distal ROI (ROI1), 10 of 11 patients with 2 measurements showed no change or a small increase in absolute bone mineral density. The results in other ROIs were similar. This longitudinal DEXA data suggest that progressive bone resorption is not problematic with an uncemented distal femur endoprosthesis at intermediate follow-up. Key words: DEXA, stress shielding, uncemented endoprothesis, tumor, bone loss. PMID- 15902862 TI - A comparison of 2 continuous passive motion protocols after total knee arthroplasty: a controlled and randomized study. AB - Effect of continuous passive motion (CPM) protocols on outcomes after total knee arthroplasty. In this prospective randomized controlled study, 147 patients were assigned to 1 of 3 treatment groups: CPM from 0 degrees to 40 degrees and increased by 10 degrees per day, CPM from 90 degrees to 50 degrees (early flexion) and gradually progressed into full extension over a 3-day period, and a no-CPM group. The CPM was administered twice a day for 3 hours over a 5-day period. All patients participated in the same postoperative physiotherapy program. Patients were assessed preoperatively, day 5, 3 months, and 1 year postoperatively. The early flexion group had significantly more range of flexion than both the standard and control groups at day 5. There was no significant difference between the groups for any other variable tested at any time frame. Key words: total knee arthroplasty, CPM, rehabilitation, outcomes. PMID- 15902863 TI - Fracture of the femur in revision hip arthroplasty with a fully porous-coated component. AB - Fifty-four consecutive femoral component revisions with uncemented fully porous coated stems were retrospectively reviewed for the prevalence of intraoperative fracture. Fracture of the femoral diaphysis occurred during insertion of a curved stem in 8 (15%) hips. One additional distal fracture of the femur occurred during exposure of the hip. All fractures occurred with a curved femoral stem. Treatment for intraoperative fractures included open reduction and internal fixation with cortical strut grafts and cables followed by protected weight bearing. This was successful in 6 hips. Even with the use of a curved stem, we report a high prevalence of intraoperative fracture of the femur in revision hip arthroplasty. PMID- 15902864 TI - The measurement of creep in ultrahigh molecular weight polyethylene: a comparison of conventional versus highly cross-linked polyethylene. AB - Quantification of creep of highly cross-linked polyethylene would enable separation of creep from wear when evaluating femoral head penetration into polyethylene. We compared creep magnitude of a highly cross-linked versus conventional polyethylene in the laboratory. Twelve acetabular liners of each material were tested, 6 of which had a 32-mm inner diameter (ID) and 6 had 28-mm ID. Creep was measured using coordinate measuring machines during loading at 2 Hz without motion to 4 million cycles. Penetration into 32-mm ID conventional liners reached 97 microm versus 107 microm for highly cross-linked material, not significant. Penetration into 28-mm conventional liners was 132 microm versus 155 microm for highly cross-linked material (P = .017). Ninety percent of the creep had occurred by 2.5 million cycles. PMID- 15902865 TI - Migration of cemented stem and restrictor after total hip arthroplasty: a radiostereometry study of 25 patients with Lubinus SP II stem. AB - The aim of this study was to investigate the migration pattern of the Lubinus SP II prosthesis stem. Migration at the stem-cement and at the bone-cement interface was assessed. Twenty-five patients were studied by radiostereometry during a period of 2 years. The migration of the head, of 2 points on the collar, of the stem tip, and also of the cement restrictor, was measured. The collar and the head were found stable at 2 years' follow-up, whereas an anterior migration occurred at the stem tip (median, 0.3 mm; 25th percentile, 0.04 mm; 75th percentile, 1.27 mm). Virtually no subsidence or axial rotation of the stems was observed. An anterior migration of the restrictor marker was also observed, compatible with a rigid sagittal plane rotation of the stem and of the cement mantle about the collar. Therefore, stem tip migration occurred at the bone cement interface. Key words: total hip arthroplasty, anatomic stem, radiostereometry, subsidence, bone-cement interface. PMID- 15902866 TI - Anatomic dimensions of the patella measured during total knee arthroplasty. AB - The anatomic measurements of 92 patellae with normal underlying bony structure were studied during total knee arthroplasty before and after resection of the articular surface. The articular surface of the patella was found to have an oval shape with a width-to-height ratio (46 x 36 mm) of 1.30. The dome was 4.8 mm high and displaced medially 3.6 mm. The medial facet was slightly thicker than the lateral facet (18 vs 17 mm). The lateral facet is 25% wider than the medial facet. Coverage provided by oval patellar prostheses was significantly better than with round prostheses. The patellae in women were significantly smaller than in men. Size differences and deformity need to be taken into account when the patella is prepared for resurfacing. It is recommended that the bony resection should be no greater than one third of the maximum patellar thickness to avoid alteration of normal bony structure. Key words: patella, total knee arthroplasty, anatomy. PMID- 15902867 TI - Effect of oral alendronate on net bone ingrowth into canine cementless total hips. AB - Alendronate has been shown to prevent osteolysis in a canine total hip arthroplasty (THA) model. However, the effect of alendronate on bone ingrowth and remodeling around canine cementless hip replacement components remains unclear. We hypothesized that oral alendronate would increase net bone ingrowth into the porous surface of THA components when intimate bone/implant contact exists. To test this hypothesis, six mature dogs received unilateral cementless THA and were treated with five milligrams of oral alendronate daily from day seven to day eighty-four postoperatively. Six comparable dogs served as controls. Quantitative analysis of net bone ingrowth into the porous surface did not show a significant difference between the two groups, (control 7.9 +/- 1.5, alendronate 7.5 +/- 1.4, P < 0.6). These results did not support the hypothesis that oral alendronate would increase net new bone formation into the porous surface of THA components. Key words: alendronate, bone ingrowth, total hip arthroplasty. PMID- 15902868 TI - [Foodborne botulism at the time of globalisation]. PMID- 15902869 TI - [Teaching the diagnosis and treatment of depressive disorders in the elderly, a front-line battle or a lost cause?]. PMID- 15902870 TI - [Epidemiology of venous thromboembolic disease]. AB - BACKGROUND: The wide variations in reported incidence and mortality rates reflect gaps in the epidemiology of venous thromboembolic disease (VTD), a clinical entity with two different manifestations--deep vein thrombosis and pulmonary embolism. We reviewed recent studies and data sources to find the most useful estimates of its epidemiologic indicators in France. METHODS: We used international medical databases to conduct a systematic literature review. Our search focused on the incidence and mortality rates of VTD in France and examined epidemiologic studies, autopsy studies, clinical trials, and national morbidity and mortality databases. RESULTS: The annual incidence of deep vein thrombosis is approximately 120 per 100,000 in France and 60-100 per 100,000 worldwide; the annual incidence of pulmonary embolism, the principal serious complication of deep vein thrombosis, is between 60 to 111 per 100,000 in France and between 23 and 107 per 100,000 internationally. The reported mortality rate for VTD in France is 7.2 per 100,000, but estimates from international autopsy series suggest a pulmonary embolism prevalence among hospital patients of 0.8 to 1%. CONCLUSION: This study demonstrates the limitations of current epidemiologic knowledge about VTD. Improved information about this disease requires better case reporting and large population-based longitudinal cohort studies. PMID- 15902871 TI - [Factors of immunodepression in patients with tuberculosis]. AB - OBJECTIVES: To study the frequency of a factor of immunodepression in patients with tuberculosis, the differences in presentation, and the diagnosis and therapeutic management according to the immune status. METHODS: Retrospective study of the files of patients hospitalised in the University Hospital Centre of Rennes in 1998 for a Mycobacterium tuberculosis infection. Comparison of two populations, immunodepressed versus non immunodepressed. RESULTS: 75 patients aged 20 to 91 were included, 41 patients were considered immunodepressed and 34 non immunodepressed. The causes of immunodepression were: HIV infection (n = 2), diabetes (n = 4), chronic alcoholism (n = 12), chronic respiratory diseases treated with corticosteroids (n = 6), neoplasia (n = 9), and inflammatory diseases (n = 7). Comparison between the 2 populations revealed more a frequent history of tuberculosis in the immunodepressed (p = 0.04), shorter delay before diagnosis (p = 0.04), greater frequency of disseminated forms (p = 0.02) and enhanced mortality (p = 0.01). There was no difference in the 2 groups with regard to the clinical signs having evoked tuberculosis, the diagnostic method, the bacteriological results or the modalities of treatment. CONCLUSION: The frequent reactivation of tuberculosis in immunodepressed patients and the severity of the infection in these patients should evoke tuberculosis and the rapid initiation of an efficient treatment in such patients. In the case of alteration in immune defences, prophylactic treatment should help to reduce the number of such reactivations. PMID- 15902872 TI - [Neisseria meningitidis infection. Clinical criteria orienting towards a deficiency in the proteins of the complement]. AB - OBJECTIVE: Complement protein deficiency of the classical pathway or in proteins of the alternate pathway is rare but considerably increase the risk of infection with Neisseria meningitidis. The aim of this study was to determine the clinical criteria of the group at risk. METHODS: Retrospective study of the clinical and biological data of patients exhibiting complement protein deficiency associated with one or several N. meningitidis infections. RESULTS: Forty cases were studied, including 35 classical pathway protein deficiencies, with a predominance of C7 deficiency, 3 properdin deficiencies and 2 acquired C3 deficiencies. More than 60% of the patients exhibited recurrent N. meningitidis infections. Serogroups of rare strains were isolated in 50% of cases. Properdin deficiency was associated with a fulminating form in 2 cases out of 3. The age at onset of the first manifestations varied from 2 months to 32 years. CONCLUSION: A deficiency must be systematically searched for in all patients presenting with a N. meningitidis infection before the age of 6 months or after the age of 5 years. Identification of deficient patients permits the proposal of family screening and appropriate prophylaxis, including preventive vaccination. PMID- 15902873 TI - [Antiretroviral therapy for HIV-infected patients with schizophrenia. Coordinated multidisciplinary management (7 cases)]. AB - OBJECTIVE: Schizophrenia might appear to be an obstacle to the initiation of and especially compliance with antiretroviral therapy for HIV-infected patients. The aims of this study were to describe the clinical, immunologic and virologic course after initiation of antiretroviral therapy in 7 HIV patients with schizophrenia (according to DSM-IV-R criteria), and to analyse the possibilities of an adequate antiretroviral therapy for those patients. OBSERVATIONS: Multidisciplinary management by specialists in infectious diseases, addiction related disorders, treatment adherence and compliance, and psychiatrists, as well as social workers, home care agencies, and patient advocacy and assistance groups, was organized with coordinated medical-psychiatric follow-up at least once a month. The patients, 6 men and 1 woman, were aged from 26 to 48 years; schizophrenia had been diagnosed in 5 patients 6 months to 20 years before the HIV infection was discovered; diagnoses of both diseases were essentially simultaneous for the other 2. All patients took long-term neuroleptics for their schizophrenia. Two were active drug addicts who received drug substitution treatment. Before antiretroviral treatment began, 6 patients had advanced infection: stage C with peak CD4 cell counts ranging from 6 to 70/mm3; they began treatment with protease inhibitors between May 1996 and August 1997. The seventh patient was first seen during primary HIV infection in July 1998, and treatment began then. Response to antiretroviral treatment with protease inhibitors was slow for all patients, but viral load became undetectable for 6 of the 7, after 5 months to 4 years; 3 had opportunistic infections. Follow-up ended in January 2002: 5 patients still had undetectable viral loads,, with CD4 cell counts ranging from 45 to 1 000/mm3. One patient died from mixed terminal cirrhosis (alcohol abuse and hepatitis C); the viral load in another was only partially controlled (10 000 copies/ml), because of poor treatment adherence. CONCLUSION: Individuals with schizophrenia can respond well to antiretroviral treatment, although response may appear slow; they can adhere to complex treatment regimens as long as they receive well coordinated and sustained multidisciplinary support. PMID- 15902874 TI - [Coexistence of dermatomyositis and macrophagic myofasciitis]. AB - INTRODUCTION: An inflammatory myopathy, characterised by joint and muscle pain, chronic asthenia, with infiltration of peri-fascicular epimysium, perimysium and endomysium by cells of the macrophagic line, macrophagic myofasciitis is often associated with other, generally auto-immune, affections. However, the coexistence with another inflammatory myopathy is relatively rare. OBSERVATION: A 29 year-old woman presented with 2 distinct inflammatory myopathies, a macrophagic myofasciitis and a dermatomyositis, which had appeared a few years after. DISCUSSION: In the rare cases in which 2 inflammatory myopathies are combined, the precise relationship between them is unknown, an individual susceptibility to developing muscle diseases is suggested. PMID- 15902875 TI - [Left auricle myxoma, a rare cause of myocardial infarction]. AB - INTRODUCTION: The association of left auricle myxoma and myocardial infarction is exceptional. Nevertheless, a causal relationship exists between the 2 affections. OBSERVATION: A 54 year-old woman was hospitalised in a rush for myocardial infarction. Sonography revealed a voluminous tumoral formation in the left auricle. The diagnosis of myxoma was confirmed by the anatomopathological examination. DISCUSSION: Although systemic embolism represents the most frequent causal link between left auricle myxoma and myocardial infarction, hypercoagulability is another possible cause. This can be explained by the secretion of interleukins 6 and 8 by the myxoma. PMID- 15902876 TI - [Ulcerating haemorrhagic colitis induced by celecoxib]. AB - INTRODUCTION: Selectivity of cyclooxygenase-2 inhibitors (Cox2 inhibitors) decreased gastroduodenal toxicity of nonsteroidal anti-inflammatory drugs. Effects on colic mucosa are little known. OBSERVATION: A case history is described of a 38-year-old women, without digestive past, who presented with hemorrhagic ulcerated acute colitis beginning 2 days after starting celecoxib (200 mg/d) prescribed for sciatica. DISCUSSION: Intrinsic imputability of celecoxib is very probable in this case report by combination of chronological and semiological criteria. Extrinsic imputability is discussed, starting from the available bibliographical data which relate primarily to rofecoxib. CONCLUSION: This observation, which constitutes, as far as we know, the first case report of hemorrhagic ulcerated colitis related to celecoxib, confirms the colic toxicity of anti-Cox2 and identify a new cause of acute colitis. Report of colic side effects with Cox2 inhibitors is required because of their new marketing. PMID- 15902877 TI - [Acute pancreatitis complicated by Clostridium tertium septicaemia]. AB - INTRODUCTION: Clostridium tertium septicaemia, although usually described in neutropenic patients, can also occur in the absence of neutropenia, as in our case report. OBSERVATION: A 61 year-old woman presented with pain in the right hypochondrium and epigastrium, constipation and vomiting that led to the diagnosis of acute pancreatitis of biliary origin. During septic shock, analysis of the liquid of an intra-abdominal mass and the results of the blood cultures revealed a Gram positive bacillus identified as C. tertium. During laparotomy, necrotic-hemorrhagic acute pancreatitis lesions were found with subtotal colic necrosis. Analysis of the peritoneal fluid revealed C. tertium. The patient died on D 46 after several nosocomial pneumopathies without identification of C. tertium. DISCUSSION: Three factors appear related to the onset of C. tertium septicaemia: neutropenia, lesions of the intestinal mucosa and previous treatment with 3rd generation, broad spectrum, cephalosporins. Such septicaemias are rare in immunocompetent patients and do not appear to have been reported before in a context of acute pancreatitis. PMID- 15902878 TI - [Internal medicine, facts and stakes in a general hospital setting]. PMID- 15902879 TI - [Botulism, a clinical diagnosis]. AB - Foodborne botulism results from the effect of a neurotoxin produced by a sporulated anaerobic bacillus called Clostridium botulinum. The mode of contamination occurs through the consumption of foodstuff, already contaminated by the neurotoxin. Following an incubation period that varies from 2 hours to 8 days, the symptoms start with intestinal problems. Then paralysis of the cranial nerve pairs sets in, classically manifested by diplopia, dysphagia, dysphonia, areactive mydriasis and ptosis. The onset of motor disorders occurs in descending order with possible involvement of the respiratory muscles, hence requiring reanimation measures and sometimes mechanical ventilation. The diagnosis of botulism is clinical. Identification of the botulinum toxin in the blood or faeces of the patients or in the contaminating food stuff confirms the diagnosis. PMID- 15902880 TI - [Foodborne botulism, epidemiologic aspects]. AB - With a mean of 30 cases reported per year, following Italy, France ranks second in the European countries in terms of incidence of botulism. Food stuff of commercial origin, of artisanal or industrial manufacture fabrication, is increasingly implicated in the genesis of outbreaks of botulism. Moreover, the modern methods of conserving food (vacuum packed food, frozen food...) allow the development of Clostridium bacteria. The diversification of the risks related to the type of products incriminated and to the new conservation methods, associated with the extension of commercial exchanges, result in the risk of widespread internationally outbreaks of botulism. PMID- 15902881 TI - [Foodborne botulism, prevent and treatment]. AB - The United States and Europe agree that the treatment of botulism is based on symptomatical measures and, notably, on mechanical ventilation when the respiratory function is impaired. Opinions diverge regarding the specific treatment represented by anti-botulinum serum: used systematically in the United States and frequently in many European countries, France never uses it other than in a few cases. Identification of the contaminating foodstuff is a fundamental element in limiting extension of the disease. Reducing the delay in declaration and the identification and correction of bad cooking practices would help to reduce the number of cases of botulism. PMID- 15902882 TI - [Utility of electroconvulsive therapy for severe depression in subjects aged over 65]. AB - The management of depression in subjects aged over 65 is based on the isolated or combined use of antidepressant chemotherapy, psychotherapy and electroconvulsive therapy. Electroconvulsive therapy, under general anaesthesia and use of curare, consists in producing a generalised seizure using a short, pulsed, electrical current administered via the transcranial route. There is renewed interest in electroconvulsive therapy with the development of specific rules and conditions for its use, together with the recruitment of depressed patients resistant to classical treatments in hospital settings. The efficacy of electroconvulsive therapy has been demonstrated in the elderly. The immediate side effects, related to the electrical stimulation and the seizure, such as headaches, nausea, confusion, and transient amnesia, regress within a few minutes or hours after the session. The limits of electroconvulsive therapy are the high risk of relapse on suspension of the sessions, relapse basically related to the severity of the depression. Consolidation electroconvulsive therapy provides new hope for better control of such relapses. PMID- 15902883 TI - [Orf nodules or "hangovers"]. PMID- 15902884 TI - Magnetic resonance cholangiopancreatography: image quality, ductal morphology, and value of additional T2- and T1-weighted sequences for the assessment of suspected pancreatic cancer. AB - PURPOSE: To assess image quality and duct morphology on magnetic resonance cholangiopancreatography (MRCP) and also the value of additional T2- and T1 weighted sequences for differentiation of benignity and malignancy in patients with suspected pancreatic tumors. MATERIAL AND METHODS: One-hundred-and-fourteen patients received MRCP and unenhanced and contrast material-enhanced MR imaging. MR results were analyzed independently by two blinded readers, and subsequently correlated with the results from surgery, biopsy, and follow-up. Assessment included the evaluation of image quality, duct visualization and morphology, and the differentiation of pancreatic lesion status (benign versus malignant). RESULTS: Overall, 49 patients had benign final diagnoses, while 65 had a malignant diagnosis. Image quality of single-shot thick-slab MRCP was rated significantly better than the MIP images of multisection MRCP. With MRCP alone, the two readers' accuracy in the assessment of pancreatic lesion status was 72% (95% CI, 64% to 83%) and 69% (95% CI, 56% to 77%), respectively; with the addition of T2- and T1-weighted images the accuracy significantly improved to 89% (95% CI, 82% to 95%) and 84% (95% CI, 77% to 92%) for readers 1 and 2, respectively. CONCLUSION: Single-shot thick-slab MRCP and multisection MRCP provide complementary results; however, single-shot MRCP had superior image quality. Moreover, assessment of ductal morphology with MRCP alone facilitated the diagnosis of different pathologic conditions of the pancreatobiliary system in the majority of patients. However, with the addition of T2- and T1-weighted sequences the overall diagnostic accuracy was significantly improved and thus we consider that a comprehensive MR approach should comprise both MRCP techniques and parenchymal sequences. PMID- 15902885 TI - Do dysphagic patients with an absent pharyngeal swallow have a shorter survival than dysphagic patients with pharyngeal swallow? Prognostic importance of a therapeutic videoradiographic swallowing study (TVSS). AB - PURPOSE: To study survival in two groups of dysphagic patients--one group unable to elicit the pharyngeal stage of swallow (APS) and another group with pharyngeal swallow (WPS)--and to compare recommendations regarding nutrition and therapeutic strategies based on the therapeutic swallowing study. MATERIAL AND METHODS: In this retrospective study, the records of dysphagic patients who have undergone a therapeutic videoradiographic swallowing study (TVSS) were reviewed. Forty patients without pharyngeal swallow were matched for age and gender with 40 patients with pharyngeal swallow; altogether 80 patients were included in the study. Survival was registered at 3, 12, and 72 months after the TVSS. RESULTS: In this study, the APS group had a significantly shorter survival time (P=0.0030) compared to the WPS group when followed-up at 12 months. In the APS group, most patients (37.5% (15/40)) died within the 3 months after TVSS. At 72 months, 62.5% (25/ 40) of the patients in the APS group had died. In the WPS group, 5% (2/40) had died within 3 months and 47.4% (19/40) after 12 months. At 72 months, 52.5% (21/40) of the patients in the WPS group had died. Regarding nutritional and therapeutic recommendations based on TVSS, 34/40 in the APS group were recommended no oral intake. Eighteen naso-gastric tubes were placed directly after TVSS. The therapeutic strategies recommended were head-positioning, thermal tactile stimulation, and tongue exercises (in 8 patients). In the WPS group, all patients were recommended oral intake. Diet modification was recommended in 29 patients. The therapeutic strategies recommended were head-positioning, thermal tactile stimulation, tongue exercises, supraglottic swallow, and effortful swallow (in 24 patients). CONCLUSION: Patients unable to elicit the pharyngeal stage of swallow had a shorter survival time than patients with pharyngeal swallow, probably due to a more severe underlying disease. Tube feeding was more frequent in the APS group. Fewer therapeutic strategies were recommended compared to the WPS group. In the WPS group, diet modification was frequent. Several patients had different therapeutic strategies. At the end of the study, 8/40 patients (20%) in the APS group had recovered and regained the ability to elicit the pharyngeal stage of swallow. All eight had achieved active swallowing rehabilitation. PMID- 15902886 TI - Sonographic comparison of gastric emptying of broth and water: is there a promoting cephalic factor? AB - PURPOSE: To examine the emptying times of broth and water, and explore the possibility of a cephalic influence on gastric emptying. MATERIAL AND METHODS: On different days each of twelve healthy volunteers had meals of either 350 ml water or 350 ml broth. Subsequently ten volunteers had meals of water alone and of water followed by sham feeding with broth. The antral area was determined at sonography five times before the meal as a baseline, and every 1-4 minutes after the meal. The time until the antral area had decreased to 150% of baseline (T150) was determined and used as surrogate expression of gastric emptying time. RESULTS: The mean T150 was for water 20.3 min (range 12-40), and was for broth significantly shorter 12.6 min (5-21), P = 0.0020. In the subsequent series the mean T150 was 28.5 min (18-49) for water, and significantly shorter for water followed by sham feeding, 22.8 (14-40), P = 0.0078. CONCLUSION: Broth empties faster from the stomach than plain water, probably because of a "cephalic phase" stimulation of gastric motility via the vagus nerve. PMID- 15902887 TI - Does computer-aided detection assist in the early detection of breast cancer? AB - PURPOSE: To evaluate whether breast cancers detected at screening are visible in previous mammograms, and to assess the performance of a computer-aided detection (CAD) system in detecting lesions in preoperative and previous mammograms. MATERIAL AND METHODS: Initial screening detected 67 women with 69 surgically verified breast cancers (Group A). An experienced screening radiologist retrospectively analyzed previous mammograms for visible lesions (Group B), noting in particular their size and morphology. Preoperative and previous mammograms were analyzed with CAD; a relatively inexperienced resident also analyzed previous mammograms. The performances of CAD and resident were then compared. RESULTS: Of the 69 lesions identified, 36 were visible in previous mammograms. Of these 36 "missed" lesions, 14 were under 10 mm in diameter and 29 were mass lesions. The sensitivity of CAD was 81% in Group A and 64% in Group B. Small mass lesions were harder for CAD to detect. The specificity of CAD was 3% in Group A and 9% in Group B. Together, CAD and the resident found more "missed" lesions than separately. CONCLUSION: Of the 69 breast cancers, 36 were visible in previous mammograms. CAD's sensitivity in detecting cancer lesions ranged from 64% to 81%, while specificity ranged from 9% to as low as 3%. CAD may be helpful if the radiologist is less subspecialized in mammography. PMID- 15902888 TI - Compliance with European guidelines for diagnostic mammography in a decentralized health-care setting. AB - PURPOSE: To evaluate the compliance of Danish mammography clinics with requirements concerning organization, activity volume, and assessment procedures from two European guidelines for quality assurance in diagnostic mammography (EUSOMA and EUREF). MATERIAL AND METHODS: We used individual records on all diagnostic mammographies performed in Denmark in 2000, and questionnaires given to Danish mammography clinics in 2000, 2002, and 2004. RESULTS: The study showed a marked centralization of the diagnostic activity from 2000 to 2004 to a smaller number of public breast assessment centers with full multidisciplinary breast assessment. However, a relatively large number of these centers did not comply with the activity volume requirement of 2000 mammograms per clinic per year. The number of private diagnostic mammography clinics performing basic diagnostic mammography has remained fairly stable in the period 2000 to 2004. Compared with public breast assessment centers, the private diagnostic mammography clinics had a lower compliance with activity volume requirements. CONCLUSION: A marked proportion of Danish public breast assessment centers operate with less than optimal activity volume, suggesting that further centralization would be appropriate. The situation in private diagnostic mammography clinics may cause concern, as our study showed that the majority of these clinics did not meet the activity volume requirements. PMID- 15902889 TI - Comparison of early and delayed quantified indices of double-phase (99m)Tc MIBI scintimammography in the detection of primary breast cancer. AB - PURPOSE: To compare the diagnostic accuracy and incremental diagnostic role of quantitative indices of early and delayed lesion to non-lesion ratios (L/Ns) in the detection of primary breast cancer. MATERIAL AND METHODS: Double-phase (99m)Tc MIBI scintimammography (SMM) (early 10 min, delayed 3 h) was performed after injection of 750 MBq of (99m)Tc MIBI in 446 highly suspected breast cancer patients (malignant: 311, benign: 135). For visual analysis, five scoring methods were used, and, for quantitative analysis, early and delayed L/Ns were calculated. Receiver operating characteristic curve (ROC) analyses were performed to determine the optimal visual grade, and to calculate cut-off values of quantitative indices for differentiation of malignant and benign diseases and to investigate whether the quantitative indices could provide incremental diagnostic values in addition to visual analysis. RESULTS: Optimal visual grades were above 4 and 5 in the detection of breast cancer. Sensitivity was 84.2% and specificity 79.3%; the area under the curve (AUC) was 0.832 (95% CI, 0.794-0.866) and standard error was 0.019. Early and delayed L/Ns of malignant breast disease were significantly higher than those of benign disease (early: 2.01 +/- 0.99 versus 1.13 +/- 0.26 (P < 0.001); delayed: 1.68 +/- 0.69 versus 1.11 +/- 0.23 (P < 0.001)). The optimal L/Ns for the detection of primary breast cancer were 1.27 for early and 1.12 for delayed imaging. When early L/N 1.27 was used as cut-off value for the detection of primary breast cancer, the sensitivity of SMM was 77.8% and specificity 85.2%. The AUC was 0.856 (95% CI, 0.820-0.888). When delayed L/N 1.12 was used, sensitivity and specificity were 81.4% and 78.5%, respectively. The AUC was 0.834 (95% CI, 0.796-0.867). The ROC comparison of early and delayed L/N showed no statistical difference in the detection of malignant breast disease (P=0.403). When the delayed L/N was added to the early one, early plus delayed quantitative analysis (E+D) showed 86.5% sensitivity and 74.8% specificity. However, the AUCs of E+D (0.854, 95% CI, 0.767-0.842) and early L/N (E) (0.856) showed no statistical difference (P=0.614). When grades 4 and 5 were used as cut-off visual grade, sensitivity and specificity were 84.2% and 79.3%, respectively. When the E was added to visual grade, visual plus early L/N (V+E) showed 89.4% sensitivity and 77% specificity. The AUC of V+E (0.867, 95% CI, 0.832 0.897) was significantly higher than that of visual analysis (V) (0.832, 95% CI, 0.794-0.866, P < 0.001). When the delayed L/N (D) was added to visual grade, visual plus delayed L/N (V+D) showed 89.4% sensitivity and 74.1% specificity. The AUCs of V+D (0.852, 95% CI, 0.816-0.884) and V revealed no statistical differences (P = 0.052). CONCLUSION: From this study, the optimal visual grades for diagnosis of breast cancer were grades 4 and 5; the cut-off values of L/Ns were 1.27 for early and 1.12 for delayed imaging. It was also found that early L/Ns provide incremental value in addition to visual analysis. However, delayed L/N revealed no incremental value. Therefore, the delayed image should not be routinely performed for purposes of primary breast cancer detection PMID- 15902890 TI - Infra-inguinal percutaneous transluminal angioplasty for limb salvage: a retrospective analysis in a single center. AB - PURPOSE: To review the feasibility of infra-inguinal angioplasty in the management of critical limb ischemia (CLI). MATERIAL AND METHODS: Data on 221 patients with 230 critically ischemic limbs, treated with consecutive percutaneous transluminal angioplasty (PTA) at Helsinki University Central Hospital between January 2000 and December 2002 were collected and analyzed retrospectively. Patency, limb salvage, and survival rates were calculated on an intention-to-treat basis. Comparisons were done with univariate (Kaplan-Meier) and multivariate analysis (Cox regression). RESULTS: Overall primary patency, secondary patency, limb salvage, and survival rates were 47%, 59%, 92%, and 76%, respectively, at 12 months. In the multivariate analysis, low toe pressure (< or =30 mmHg) was a significant risk factor for poor patency. Uremia with hemodialysis, low toe pressure (< or =30 mmHg), and hemodynamic failure of the endovascular procedure were found to increase significantly the risk of amputation. Uremia with hemodialysis, coronary artery disease, tissue loss as indication for PTA (Fontaine stage IV), and age over 70 years were all found to increase significantly the risk of death. CONCLUSION: Infra-inguinal PTA is feasible in patients with CLI and resulted in good limb salvage. PMID- 15902891 TI - Vessel size estimation in peripheral artery interventions: are angiographic measurements reliable? AB - PURPOSE: To describe and evaluate a new technique for angiographic measurement of arterial diameters and to make a comparison with other common techniques. MATERIAL AND METHODS: The Angiometer has a radiopaque metal body with six cylindrical segments of varying diameter. Arterial diameter is estimated by visually relating the vessel to these segments. The measurements (method A) were compared: to measuring in relation to a ruler on the table-top by means of compasses (B), to an automated measurement technique (quantitative angiography) utilizing calibration to a catheter of known diameter (C), and to a computer based distance measurement using the ruler on the table-top as reference (D). Twenty-five patients were studied. Each method was used twice by each of two independent observers. A phantom with four drilled holes, filled with contrast agent, was also studied. RESULTS: The coefficient of variation was highest for method C and lowest for method A. Correlations between readings were highest for method A and lowest for method C. Mean diameter values were highest for method A and lowest for methods B and D. In the phantom experiments, sizes were overestimated by between 5% and 9% with method A, and underestimated by between 4% and 27% with the other methods. CONCLUSION: In terms of reproducibility and accuracy, the proposed method compares favorably with alternative methods. PMID- 15902892 TI - Three-dimensional rotational angiography of transplanted renal arteries: influence of an extended angle of rotation on beam-hardening artifacts. AB - PURPOSE: To investigate whether three-dimensional rotational angiography (3D-RA) of the transplant renal artery performed with an extended angle of rotation can reduce beam-hardening artifacts in 3D reconstructed images without image quality being lost or side effects to the transplanted kidney being increased. MATERIAL AND METHODS: 3D-RA with a C-arm rotation of 180 degrees was performed consecutively in 12 renal transplanted patients with suspicion of renal artery stenosis. A 1.7-mm balloon occlusion catheter was placed using the crossover technique and this was compared to a protocol with 160 degrees rotation and a traditional 1.4-mm catheter in 10 patients. The occurrence of beam-hardening artifacts was registered and the effects of the reduced contrast load on image quality and of arterial occlusion on renal function were assessed. RESULTS: The extended angle of rotation, from 160 degrees to 180 degrees, reduced the beam hardening artifacts. Artifacts were observed in 4/11 patients (36%) in the study group and in all 10 (100%) of the controls. There was no statistical difference regarding image quality between the two protocols. Renal function was equally affected in both protocols. CONCLUSION: 3D-RA with an extended C-arm rotation reduced the beam-hardening artifacts. Image quality was not reduced despite the reduced contrast medium load. The different protocols had no effect on patient outcome. PMID- 15902893 TI - Value of multidetector computed tomography in assessing blunt multitrauma patients. AB - PURPOSE: To find out if multidetector computed tomography (MDCT), using a dedicated trauma protocol, provides sufficient diagnostic information of the injuries of blunt multitrauma patients to enable the planning of treatment for all body compartments. MATERIAL AND METHODS: One-hundred-and-thirty-three patients exposed to high-energy trauma were referred and scanned with the standardized MDCT multitrauma protocol. The imaging protocol consisted of axial scanning of the head and helical scanning of the facial bones, cervical spine, thorax, abdomen, and pelvis. The scanning times were 12 s for the head, 19-21 s for the facial bones and cervical spine (1 mm collimation), and 32-50 s for the thorax, abdomen, and pelvis (2 mm collimation). One-hundred-and-forty milliliters of non-iodinated contrast material (300 mg I/ml) was administered intravenously at 3 ml/s. RESULTS: Ninety-nine of the patients (74%) had at least one finding consistent with trauma. The most frequent findings were in the thorax in 58 patients (44%). Nineteen false-negative findings and two false-positive findings were made. The overall sensitivity of MDCT was 94%, specificity 100%, and accuracy 97%. CONCLUSION: MDCT is accurate in the assessment of blunt multitrauma patients. The decision to treat the patient can be made on the basis of MDCT with a reasonable level of certainty. PMID- 15902894 TI - Cerebellar hematoma following transsphenoidal surgery. AB - Cerebellar hemorrhage after supratentorial surgery is a very infrequent complication. Most cases occur following aneurysm or brain tumor surgery, or evacuation of extracerebral collections. The underlying mechanism of formation of these hematomas is unclear. We present an example of cerebellar hemorrhage following transsphenoidal resection of a pituitary adenoma. To our knowledge, this is the first case reported of hemorrhage in the posterior fossa after a transsphenoidal approach. PMID- 15902895 TI - Magnetic resonance findings of primary central nervous system T-cell lymphoma in immunocompetent patients. AB - PURPOSE: To describe the MR findings of primary central nervous system T-cell lymphoma (T-PCNSL) in immunocompetent patients. MATERIAL AND METHODS: Seven patients with pathologically proven T-PCNSL were included in our study. The number, location, shape, enhancement pattern, and signal intensity of the tumors were determined. Diffusion-weighted images (DWI) and perfusion-weighted images (PWI) were obtained in four and two patients, respectively. Apparent diffusion coefficients (ADCs) were generated, and regions of interest were defined in each lesion. RESULTS: Four patients with T-PCNSL had a single mass, while the others had multiple lesions (four, three, and two lesions, respectively). All seven cases of T-PCNSL had a supratentorial location: 12 in the subcortical area and 1 in the thalamus. No leptomeningeal involvement was noted. All tumors showed iso- to low T1 and iso- to slightly high T2 signal intensity to the adjacent gray matter. Rim enhancement was seen in 5 of the 7 patients (71.4%), while heterogeneous and homogeneous enhancement was seen in each of two. On DWI and ADC maps, the enhancing lesions showed slight hyperintensity in three patients (mean ADC ratio, 0.92 +/- 0.06) and iso-intensity in the other (ADC ratio, 1.02 +/- 0.05). Cystic areas consistent with necrosis were noted in three patients. High signal intensity area in the cortex was noted on T1-weighted images in three patients, suggesting hemorrhage. In two patients, the same signal intensity area was noted within the mass. The two masses on the relative cerebral blood volume (rCBV) map demonstrated either similar or slightly higher signal intensity than that of the contralateral white matter. The rCBV ratios of these two masses were 1.27 +/- 0.16 and 1.35 +/- 0.2, respectively. CONCLUSION: T-PCNSLs show a predilection for a subcortical location, a relatively high incidence of cortical or intratumoral hemorrhage, rim enhancement, or cystic-areas consistent with necrosis on magnetic resonance imaging. The lower rCBV ratio of the tumor might be helpful in differentiating T-PCNSL from other brain tumors such as high-grade glioma. PMID- 15902896 TI - Posterior "nutcracker" phenomenon: hemodynamic relevant aorto-retroaortal renal vein fistula leading to fatal right heart failure. AB - A rare complication of an aneurysm of the abdominal aorta is the so-called "posterior nutcracker phenomenon", which describes compression of a retroaortal renal vein between the abdominal aorta and the vertebral column. The clinical presentation is flank pain and hematuria, which are usually caused by a renal (respectively ureteral) calculus or neoplasia. Another rare differential diagnosis for these very common clinical symptoms is an aorto-left renal vein fistula (ALRVF), which is a spontaneous vascular fistula, usually also associated with an aortic aneurysm, infrequently a result of abdominal trauma. PMID- 15902897 TI - Diffusion-weighted and conventional MR imaging in neurotrichinosis. AB - Central nervous system involvement in trichinosis is not rare. Brain lesions in trichinosis have been defined on computed tomography and magnetic resonance imaging (MRI) as multifocal small lesions located in the cerebral cortex and white matter. We present a case of trichinosis with multifocal lesions of the brain detected by MRI and diffusion weighted MRI. Evolutions of these lesions from acute through chronic stages on follow up studies are also presented. This is the first report describing sequential MRI findings and diffusion weighted imaging appearance of brain lesions in trichinosis. Sequential evaluation of conventional and diffusion MR data allowed us to conclude that multifocal lesions in the brain were related to multiple infarctions rather than true inflammatory infiltration of the brain parenchyma. PMID- 15902898 TI - Metacarpophalangeal pattern profile analysis in Leri-Weill dyschondrosteosis. AB - PURPOSE: To analyze the metacarpophalangeal profile (MCPP) in individuals with Leri-Weill dyschondrosteosis (LWD) and to assess its value as a possible contributor to early diagnosis. MATERIAL AND METHODS: Hand profiles of 39 individuals with a diagnosis of LWD were calculated and analyzed. Discriminant analysis was applied to differentiate between LWD and normal individuals. RESULTS: There was a distinct pattern profile in LWD. Mean pattern profile showed two bone-shortening gradients, with increasing shortening from distal to proximal and from medial to lateral. Distal phalanx 2 was disproportionately long and second metacarpal was disproportionately short. Discriminant analysis yielded correct classification in 72% of analyzed cases. CONCLUSION: MCPP is not age related and the analysis can be applied at any age, facilitating early diagnosis of LWD. In view of its availability, low costs, and diagnostic value, MCPP analysis should be considered as a routine method in the patients of short stature where LWD is suspected. PMID- 15902899 TI - Percutaneous radiofrequency ablation of renal cell carcinoma: preliminary results. AB - PURPOSE: To report early results in percutaneous radiofrequency ablation (RFA) of renal cell carcinoma with an expandable RF probe. MATERIAL AND METHODS: In 14 patients (9 male, mean age 67.9 +/- 9.9 years) CT-guided percutaneous radiofrequency ablation of 15 renal cell carcinomas was performed using an expandable LeVeen probe (diameter 2-4 cm) and a 200-watt generator under general anesthesia and CT control. Tumors exceeding a diameter of 3 cm (n=6) were embolized within 24 h prior to RFA. Average tumor size was 3.0 +/- 1.0 cm. RESULTS: RFA was technically successful in all patients, resulting in a mean size of necrosis of 3.7 +/- 0.7 cm. With the exception of one reno-cutaneous fistula, which was successfully treated conservatively, no major complications were observed. No local recurrence was observed (follow-up: 13.9 +/- 12.4 months) while extrarenal tumor progression occurred in four patients. CONCLUSION: Our preliminary data suggest that nephron sparing percutaneous RFA of renal tumors with an expandable RF probe is safe and effective. PMID- 15902900 TI - The use of banana in MRCP. PMID- 15902901 TI - Pathology of chronic constipation in pediatric and adult coloproctology. AB - In colonic motility disorders, a pathohistological diagnosis based solely on formalin-fixed gut is often inconclusive. Classical histological techniques or immunohistochemistry represent a static staining. In contrast, native tissue submitted to enzyme histochemistry provides functional information about the effectiveness of the cellular performance. Routinely, a complementary set of reactions is performed and includes acetylcholinesterase (AChE), lactic and succinic dehydrogenase, as well as nitroxide synthase reactions. In this monograph, the whole spectrum of different anomalies of the colonic wall is illustrated in a systematic fashion: Hirschsprung's disease is characterized by an increase in AChE activity of parasympathetic nerve fibers of the rectosigmoid. In ultrashort Hirschsprung's disease, only enzyme histochemistry renders a reliable diagnosis possible in biopsies of the anal ring. Aganglionosis of the musculus corrugator cutis ani shows a localized increase of AChE activity in nerve fibers, similar to Hirschsprung's disease, not detectable in conventional histology. Immaturity, hypoganglionosis and neuronal dysganglionosis can be clearly recognized in dehydrogenase reactions. Enzyme histochemical reactions are complemented by picrosirius red staining for assessment of the collagen texture of the muscularis propria. Absence or intertenial interruption of the continuous connective tissue layer between circular and longitudinal muscle of the muscularis propria has been termed aplastic or atrophic desmosis, respectively. Many of the entities described are also observed in adults. Atrophic hypoganglionosis or atrophic desmosis with loss of the myenteric plexus connective tissue fascia is implied as a frequent cause of chronic constipation in adults. The essential contribution of a functional histopathological technique towards a reliable diagnosis of gut dysfunction in native tissue is extensively demonstrated in great detail in more than two hundred figures. PMID- 15902902 TI - Fyn kinase does not reduce ethanol inhibition of zinc-insensitive NR2A-containing N-methyl-D-aspartate receptors. AB - N-methyl-D-aspartate (NMDA) receptors are ion channels activated by the neurotransmitter glutamate and are important mediators of neuronal signal transduction. Ethanol inhibits ion flux through NMDA receptors at concentrations that are associated with behavioral signs of intoxication. The overall sensitivity of NMDA receptors to ethanol is influenced by factors, including subunit composition and interactions with cytoskeletal elements. Results of studies also support the suggestion that the ethanol inhibition on NR1/2A receptors is reduced by Fyn kinase-mediated tyrosine phosphorylation. However, tyrosine kinases also reduce the high-affinity zinc sensitivity of NR1/2A receptors, supporting the suggestion that kinase-dependent effects on ethanol inhibition may be secondary to relief of zinc inhibition. In the current study, the effect of Fyn kinase on the ethanol inhibition of NR1/2A receptors was determined under conditions in which zinc sensitivity is eliminated. Human embryonic kidney 293 (HEK 293) cells were transiently transfected with wild-type or mutant NMDA subunits, and glutamate-activated currents were measured by using patch-clamp electrophysiology. Inclusion of the tyrosine phosphatase inhibitor potassium bisperoxo(1,10-phenanthroline)oxovanadate (V) [bpV(phen)] in the recording pipette eliminated the potentiation of NR1/2A currents by heavy metal chelators. Under these conditions, Fyn kinase did not reduce ethanol inhibition of wild-type receptors. Fyn kinase also had no effect on the magnitude of ethanol inhibition of zinc-insensitive NR1/2A(H128S) receptors. Together, results of the current study indicate that Fyn kinase does not directly affect the ethanol sensitivity of NR1/2A receptors. PMID- 15902903 TI - Nerve growth factor produced by activated human monocytes/macrophages is severely affected by ethanol. AB - Ethanol intake impairs immune function and increases the incidence of infection in the host. Although the precise cellular target of this immunotoxic action is still unknown, findings of several studies have shown that ethanol acts on the immune response predominantly by interfering with the ability of blood monocyte derived macrophages to produce cytokines and growth factors. Nerve growth factor (NGF) represents a key molecule in monocyte/macrophage-mediated responses. Thus, we tested the hypothesis that exposure to ethanol would affect NGF synthesis as well as expression of NGF receptor trkA in this cell population. Because NGF has been reported to affect the synthesis of proinflammatory cytokines, we also evaluated whether the production of tumor necrosis factor-alpha would be affected by ethanol-mediated changes in NGF synthesis. The study results demonstrated that the acute exposure of lipopolysaccharide-activated human monocyte/macrophage cultures to ethanol led to a sharp decrease in endogenous-produced NGF, which is associated with a reduced expression of high-affinity NGF receptor on cell membrane, and to a concomitant impairment of tumor necrosis factor-alpha production. Taken together, the current findings support the suggestion that a new mechanism exists by which ethanol can compromise the efficiency of the mononuclear phagocyte system in dealing with infection and host inflammatory response. PMID- 15902904 TI - Association of polymorphisms in nicotinic acetylcholine receptor alpha 4 subunit gene (CHRNA4), mu-opioid receptor gene (OPRM1), and ethanol-metabolizing enzyme genes with alcoholism in Korean patients. AB - Findings obtained from several studies indicate that ethanol enhances the activity of alpha4beta2 neuronal nicotinic acetylcholine receptor and support the possibility that a polymorphism of the nicotinic acetylcholine receptor alpha4 subunit gene (CHRNA4) modulates enhancement of nicotinic receptor function by ethanol. To identify the association between the CfoI polymorphism of the CHRNA4 and alcoholism, we examined distribution of genotypes and allele frequencies in Korean patients diagnosed with alcoholism (n = 127) and Korean control subjects without alcoholism (n = 185) with polymerase chain reaction-restriction fragment length polymorphism methods. We were able to detect the association between the CfoI polymorphism of the CHRNA4 and alcoholism in Korean patients (genotype P = .023; allele frequency P = .047). The genotypes and allele frequencies of known polymorphisms in other alcoholism candidate genes, such as alcohol metabolism related genes [alcohol dehydrogenase 2 (ADH2), aldehyde dehydrogenase 2 (ALDH2), alcohol dehydrogenase 3 (ADH3), and cytochrome P450 2E1 (CYP2E1)] and mu-opioid receptor gene (OPRM1), were studied. The polymorphisms of ADH2, ALDH2, and CYP2E1 were significantly different in Korean patients with alcoholism and Korean control subjects without alcoholism, but ADH3 and OPRM1 did not differ between the two groups. PMID- 15902905 TI - First-pass metabolism of ethanol in human beings: effect of intravenous infusion of fructose. AB - Intravenous infusion of fructose has been shown to enhance reduced form of nicotinamide adenine dinucleotide reoxidation and, thereby, to enhance the metabolism of ethanol. In the current study, the effect of fructose infusion on first-pass metabolism of ethanol was studied in human volunteers. A significantly higher first-pass metabolism of ethanol was obtained after administration of fructose in comparison with findings for control experiments with an equimolar dose of glucose. Because fructose is metabolized predominantly in the liver and can be presumed to have virtually no effects in the stomach, results of the current study support the assumption that only a negligible part of first-pass metabolism of ethanol occurs in the stomach. PMID- 15902906 TI - Effect of ethanol on 24-hour hormonal changes in peripubertal male rats. AB - We analyzed the effect of chronic (4 weeks) ethanol feeding on 24-h variation of pituitary-testicular function in peripubertal male Wistar rats by measuring circulating concentrations of prolactin, follicle-stimulating hormone, luteinizing hormone, testosterone, and thyrotropin. Animals were maintained under a 12-h light: 12-h dark photoperiod and received a liquid diet for 4 weeks, starting on day 35 of life. The ethanol-fed group received a diet similar to that provided to control animals, except that maltose was replaced isocalorically with ethanol. Ethanol replacement provided 36% of the total caloric content of the diet. Rats were killed at one of six times around the clock, beginning at zeitgeber time (ZT) 1 (ZT 0 = lights on). In ethanol-fed rats globally, secretion of prolactin was augmented, whereas secretion of follicle-stimulating hormone, luteinizing hormone, testosterone, and thyrotropin was decreased. Significant changes in the 24-h secretory pattern of circulating hormones occurred in rats receiving ethanol, including the appearance of two peaks (at ZT 1 and ZT 9), rather than one peak, of follicle-stimulating hormone during the inactive phase of the daily cycle, suppression of the maximum plasma luteinizing hormone concentration during the first part of the inactive phase, and appearance of a second peak of testosterone and prolactin during the second part of the inactive phase (at ZT 5 and ZT 9, respectively) and of a second peak of plasma thyrotropin during the first part of the active phase (at ZT 13). The significant positive correlation between testosterone and individual luteinizing hormone and prolactin concentrations in control animals was no longer observed after ethanol administration. Chronic ethanol administration presumably affects the endogenous clock that modulates the circadian variation of the pituitary-gonadal axis and thyrotropin release in growing male rats. PMID- 15902907 TI - Impulsivity in abstinent alcohol-dependent patients: relation to control subjects and type 1-/type 2-like traits. AB - Extensive literature has linked behavior control problems in childhood to risk for alcoholism, but impulsivity in alcohol-dependent adults has not been well characterized. Using a variety of laboratory measures of impulsivity, we assessed whether detoxified alcohol-dependent patients [(ADP); n = 130] were more impulsive than control subjects [(CS); n = 41]. In comparison with CS, ADP demonstrated (1) increased rates of commission errors, but not omission errors, in a continuous performance test, (2) a more severe devaluation of delayed reward, (3) increased rates of risky responses in a new risk-taking paradigm, and (4) higher psychometric scores of impulsivity and aggression. Across all subjects, aggressiveness correlated significantly with severity of delay discounting. A post hoc analysis of data obtained for male ADP indicated that, in comparison with patients with late onset of problem drinking and no problem drinking parent, those ADP with earlier age of problem drinking and who reported a problem-drinking father (type 2-like alcohol dependence) demonstrated faster response latencies and more responses to non-target stimuli (commission errors) in the continuous performance test, as well as higher psychometric aggression. In contrast, these subtypes of male ADP did not differ in delay discounting and risk taking. These findings collectively indicate that, in comparison with CS, ADP are more impulsive in several dimensions, with elevated impulsivity in a working memory task as well as aggressivity characteristic of alcohol-dependent men with type 2-like features. PMID- 15902908 TI - Fatty acid ethyl esters: markers of alcohol abuse and alcoholism. AB - Chronic alcoholism, which is associated with hepatic, pancreatic, and myocardial diseases, is one of the major health problems in the United States with high morbidity and mortality. Many individuals who abuse alcohol chronically die even before reaching the clinical stage of the disease. Reliable biomarkers of the diseases induced by chronic alcohol abuse, as well as for alcoholism, currently are not available. In the current study, we measured plasma concentrations of fatty acid ethyl esters [(FAEEs), nonoxidative metabolites of ethanol] in 39 patients with a detectable concentration of alcohol in their blood samples. In turn, we determined the relation of FAEE concentrations with blood alcohol concentration (BAC). Of 39 patients in whom we evaluated this relation, only five had a history of chronic alcohol abuse, and six had a history of acute alcohol abuse. Patients' age ranged from 25 to 71 years. Within this age range, greater concentrations of FAEEs were found in the plasma samples obtained from patients in the 41- to 50-year age group. There were no sex-related differences in BAC, nor in FAEE concentrations. Thirteen patients had a BAC greater than 300 mg%. For 11 patients, the BAC ranged between 200 and 299 mg%, and, for 12 patients, the BAC ranged between 100 and 199 mg%. In comparison with findings for patients with a BAC that ranged between 100 and 299 mg%, the FAEE concentrations were approximately twofold higher in patients with a BAC greater than 300 mg%. Ethyl palmitate and ethyl oleate were the main FAEEs detected in most patients. In general, FAEE concentrations increased with increasing BAC. However, in comparison with patients with a history of acute alcohol abuse, a greater increase in total FAEE concentrations was observed in patients with a history of chronic alcohol abuse (4,250 ng/ml and 15,086 ng/ml, respectively). Fatty acid ethyl esters were either detected in trace amounts or not detectable in the plasma of control subjects with no known alcohol ingestion. These results support our hypothesis that nonoxidative metabolism of ethanol to FAEEs is an important pathway of ethanol disposition during chronic alcohol abuse, and that FAEE concentrations can be a more reliable biomarker of chronic alcohol abuse than a history of acute alcohol abuse. PMID- 15902909 TI - Ethanol consumption improves avoidance learning in rats: role of deprivation interval. AB - Voluntary oral self-administration of ethanol in rats has been used to model ethanol consumption and abuse in human beings, with contradictory results. The purpose of the current study was to assess the effect of voluntary ethanol consumption on acquisition of a lever press escape/avoidance task in rats. Male Wistar rats were exposed to ethanol in a limited-access procedure, and either 1 day or 10 days after their last ethanol exposure, animals received a 4-h lever press escape/avoidance session. Control animals were not exposed to ethanol at any time. Animals in the 1-day ethanol-deprivation group performed significantly better than did the other two groups with respect to avoidance responding. There were no group differences in number of lever presses during a safety period, a measure of anxiety. Further, we obtained a significant negative correlation between behavioral performance and change in ethanol consumption after the escape/avoidance session, as well as a significant positive correlation between baseline ethanol consumption and avoidance performance. Results are discussed in terms of the potential neural mediators of the improved avoidance effect in animals in the 1-day ethanol-deprivation group. PMID- 15902910 TI - Substitution profiles of N-methyl-D-aspartate antagonists in ethanol discriminating inbred mice. AB - C57BL/6J (B6) and DBA/2J (D2) inbred mice show pronounced differences in ethanol induced behaviors, such as loss of righting reflex and locomotor activation, among others. They also differ in measures of conditioned place preference and oral self-administration of ethanol. In the current study, I examined whether B6 and D2 mice differed in their expression of the N-methyl-D-aspartate (NMDA) receptor-mediated component of the discriminative stimulus effects of ethanol. B6 and D2 mice were trained to discriminate ethanol (1.5 g/kg, i.p.) from saline in a two-choice, milk-reinforced operant procedure. After training was completed, substitution and response rate dose-effect curves were generated for ethanol; the uncompetitive NMDA antagonists phencyclidine and ketamine; and the competitive NMDA antagonist D-CPPene. Dose-effect curves were also generated for midazolam, cocaine, m-chlorophenylpiperazine (mCPP), morphine, and gamma-hydroxybutyric acid (GHB). B6 and D2 mice learned the ethanol-versus-saline discrimination. Phencyclidine produced near full substitution for ethanol in both strains, whereas ketamine fully substituted for ethanol only in B6 mice. D-CPPene partially substituted for ethanol in both strains. Moderate doses of phencyclidine produced greater response rate-increasing effects in B6 mice than in D2 mice, and high doses of phencyclidine were more potent for suppressing response rates in D2 mice. In contrast, D-CPPene had similar response rate increasing effects in both strains, but high doses produced more potent response rate-decreasing effects in B6 mice. Among the other drugs tested, only midazolam produced substantial substitution for ethanol. Taken together, these findings seem to indicate that the behavioral effects of NMDA antagonists differ between strains, but that the NMDA-mediated component of the discriminative stimulus effects of ethanol is similar in B6 and D2 mice. PMID- 15902911 TI - Effects of periadolescent ethanol exposure on alcohol preference in two BALB substrains. AB - Ethanol exposure during adolescence is a rite of passage in many societies, but only a subset of individuals exposed to ethanol becomes dependent on alcohol. To explore individual differences in response to ethanol exposure, we compared the effects of periadolescent ethanol exposure on alcohol drinking in an animal model. Male and female mice of two BALB substrains were exposed to ethanol in one of three forms--choice [water vs. 10% (volume/volume) ethanol], forced (10% ethanol in a single bottle), or gradual (single bottle exposure, starting with 0.5% ethanol and increasing at 2-day intervals to 10% ethanol)--from the 6th through the 12th week of age and administered two-bottle alcohol preference tests (10% ethanol vs. water) for 15 days immediately thereafter. All three forms of ethanol exposure increased alcohol preference in male and female BALB/cByJ mice, relative to findings for ethanol-naive control animals. Only gradual ethanol exposure produced an increase in alcohol preference in BALB/cJ mice. During extended alcohol preference testing (for a total of 39 days) of mice in the gradual ethanol exposure group, the higher alcohol preference of the gradual ethanol-exposed BALB/cByJ male mice persisted, but alcohol preference of control group female mice in this strain--formerly ethanol naive, but at this point having received 10% ethanol in the two-bottle paradigm for 15 days--rose to the level of alcohol preference of female mice in the gradual ethanol exposure group. This finding demonstrated that both adolescent and adult ethanol exposure stimulated alcohol preference in female mice of this strain. Across days of testing in adulthood, alcohol preference of the gradual ethanol-exposed BALB/cJ mice decreased, resulting in a lack of effect of gradual exposure to ethanol on alcohol preference in both male and female mice of this strain during the period of extended testing. These strain differences support a genetic basis for the effects of ethanol exposure on alcohol preference and fit within a body of literature, showing substantial individual differences in the effects of ethanol exposure among genetically undefined rats and differences in response to ethanol exposure among inbred rat strains. Exploration of the mechanisms underlying this gene by environment interaction in a mouse model may help elucidate individual differences in the effects of ethanol exposure in human beings and contribute to the understanding of the causes of alcoholism. PMID- 15902912 TI - Neurophysiologic consequences of neonatal ethanol exposure in the rat. AB - Many of the neurotoxic and neurobehavioral consequences of neonatal ethanol exposure in the rat have been characterized. However, in few studies has adult neurophysiologic function been assessed in rats exposed to ethanol during this key developmental period. In the current study, the effects of neonatal ethanol exposure on adult electroencephalographic (EEG) activity and auditory event related potentials (ERPs) were examined in the rat. Male Sprague-Dawley rats were exposed to ethanol at 6.0 g/kg/day between postnatal days 4 through 9 by using an artificial-rearing procedure. Two control groups were used: a suckle control (SC) group and a gastrostomized control (GC) group. After reaching adulthood (i.e., at 3.5-4 months old), recording electrodes were implanted into the brain of each rat, so that EEG activity and auditory ERPs from the cortex and hippocampus could be assessed. Rats exposed to ethanol during the neonatal period were hyperactive as adults. Assessment of the EEG activity revealed that ethanol exposure increased peak frequency in the frontal cortical and parietal cortical 16-32 Hz frequency bands. Assessment of ERPs revealed that parietal cortical N1 amplitude was reduced in ethanol-exposed rats. Furthermore, parietal cortical N1 latency was increased in the GC group. These findings demonstrate that enhanced motor activity in rats exposed to ethanol during neonatal development occurs in combination with EEG indices of enhanced cortical and hippocampal arousal. Furthermore, a deficiency in cortical N1 amplitude indicates adult rats may have attention deficits. Overall, these results indicate that neonatal ethanol exposure has enduring neurobehavioral consequences, which persist into adulthood. This neurobehavioral profile in the rat is consistent with clinical observations of attention deficits and hyperactivity in children exposed to ethanol during prenatal development. PMID- 15902913 TI - A high-fat meal or injection of lipids stimulates ethanol intake. AB - Findings of earlier studies support the idea of a possible relation between dietary fat and ethanol intake, but it is unclear whether acute exposure to fat can increase ethanol consumption directly. In the current series of experiments, we examined whether daily overeating of fat, a single high-fat meal, or the injection of fat can increase ethanol intake. In Experiment 1, adult Sprague Dawley rats were maintained on a high-fat diet (50% fat) for 7 days and switched subsequently to a laboratory chow diet while being trained to drink 9% ethanol. Rats that had eaten the greatest amount of the high-fat diet subsequently drank the most ethanol. In Experiment 2, a 1-h meal of the high-fat diet (50% fat) produced a significant increase in 7% ethanol consumption in comparison with what occurred after consumption of an equicaloric, low-fat (10% fat) meal. In Experiment 3, the orosensory effect of fat was eliminated with an intraperitoneal injection of a fat emulsion, Intralipid (20% fat, 5.0 ml). The injection of Intralipid, in comparison with saline, increased the ingestion of 9% ethanol. This finding is in contrast to what occurred with injection of an equicaloric, 50% glucose solution, which suppressed ethanol intake. These findings provide new evidence to support a positive relation between dietary fat and the consumption of ethanol. PMID- 15902914 TI - Sugar-dependent rats show enhanced intake of unsweetened ethanol. AB - Rats show signs of dependence on sugar when it is available intermittently, including bingeing, withdrawal, and cross-sensitization with amphetamine. In the current study, we sought to determine whether sugar-dependent rats would show increased intake of unsweetened ethanol and, conversely, whether intermittent access to ethanol would augment sugar consumption. In Experiment 1, with intermittent versus ad libitum access to ethanol, Sprague-Dawley rats were given escalating concentrations of ethanol (1%, 2%, 4%, 7%, and 9%) over the course of 20 days. Rats in the intermittent ethanol access group, with 12-h daily access, consumed more 4%, 7%, and 9% ethanol during the first hour of access, and more 9% ethanol daily, than did rats in the ad libitum ethanol access group. In Experiment 2, with ethanol as a gateway to sugar intake, the rats from Experiment 1 were switched to 10% sucrose with 12-h daily access for 1 week. Rats in the intermittent ethanol access group consumed significantly more sugar than was consumed by rats in a control group with no prior ethanol experience. In Experiment 3, with sugar as a gateway to ethanol to determine whether sugar dependence leads to increased ethanol intake, four groups were maintained for 21 days according to the following designations: intermittent access to sugar and chow, ad libitum access to sugar and chow, intermittent access to chow, or ad libitum access to chow. Four days later, all groups were switched to intermittent ethanol access, as described in Experiment 1. The group with intermittent access to sugar and chow consumed the most 9% ethanol, supporting the suggestion that sugar dependence alters a rat's proclivity to drink ethanol. These results may relate to the co-morbidity between binge-eating disorders and alcohol intake and the tendency of people abstaining from alcohol to consume excessive amounts of sugar. In conclusion, bingeing on either ethanol or sugar fosters intake of the other. PMID- 15902915 TI - Plasma homocysteine concentrations do not influence craving in alcohol withdrawal. AB - Results of a number of studies indicate that the glutamate system, especially the N-methyl-D-aspartate (NMDA) receptor, has a major function in chronic alcoholism, including craving. Homocysteine and other excitatory amino acids, such as glutamate and aspartate, lead to an overstimulation of NMDA receptors. Because alcoholism is associated with elevated plasma homocysteine concentrations, we designed the current study to determine whether elevated plasma homocysteine concentrations have an influence on craving in alcohol withdrawal. Two groups of patients with an established diagnosis of alcohol dependence were compared. Group A comprised 50 consecutively admitted alcohol-dependent individuals who had been abstinent from alcohol between 24 and 72 h before hospitalization. Group B comprised 146 consecutively recruited alcohol-dependent individuals who were admitted, acutely intoxicated, for withdrawal treatment. All patients were assessed with the Obsessive Compulsive Drinking Scale (OCDS) on the day of admission and after 7 days of treatment. The mean (27.1, S.D. 18.4) plasma homocysteine concentration for group B was significantly higher than the mean (12.5, S.D. 5.3) plasma homocysteine concentration for group A (Mann-Whitney U test: P < .001). No significant influence of homocysteine concentration on the extent of craving was found for either group with the use of the Spearman correlation (day 0: group A, r = -.076, P = .601; group B, r = .120, P = .148) and logistic regression analysis. Although homocysteine is a potent modulator of glutamatergic neurotransmission, results of the current study provide no evidence for a pathophysiologic role of homocysteine in withdrawal craving. Therefore, further research about alcohol craving should focus on neurobiologic factors other than homocysteine. PMID- 15902916 TI - Formation of ethyl nitrite in vivo after ethanol administration. AB - The purpose of the current study was to ascertain whether ethyl nitrite could be detected in vitro from the reaction of ethanol with peroxynitrite, as well as after administration of ethanol to mice. Ethyl nitrite analyte was determined by using gas chromatography--mass spectrometry with headspace analysis with the use of a solid-phase microextraction device. Peroxynitrite was allowed to react with ethanol under a variety of conditions in vitro. Ethyl nitrite was generated when peroxynitrite was allowed to react with ethanol. Male, inbred short-sleep mice were injected intraperitoneally with either ethanol [5.2 g/kg; 15.0% (weight/volume) ethanol in saline] or a 50:50 mixture of deuterium-labeled ethanol (D5-ethanol) and ethanol. Blood samples, as well as whole brain and liver sections, were obtained from mice 30 min later for determination of ethanol, D5 ethanol, ethyl nitrite, and deuterium-labeled ethyl nitrite (D5-ethyl nitrite). Time courses for the appearance of ethyl nitrite in blood samples, as well as in whole brain and liver sections, obtained from mice were carried out. After ethanol administration, ethyl nitrite was detected and quantitated in mouse blood, brain, and liver. A small fraction of ethyl nitrite was present. When a 50:50 mixture of ethanol and D5-ethanol was given to animals, both ethyl nitrite and D5-ethyl nitrite were found in blood and brain in approximately the same ratio as that of ethanol and D5-ethanol. The level of D5-ethyl nitrite in liver was more than twice that of ethyl nitrite, indicating a possible isotope effect in the metabolism of ethyl nitrite. Ethyl nitrite is a new metabolite of ethanol in vivo. The mechanism of ethyl nitrite formation is most likely the reaction of ethanol with peroxynitrite generated in vivo from nitric oxide. PMID- 15902917 TI - Effects of acute and short-term administration of tryptophan plus ethanol on noradrenaline and serotonin metabolites in the locus coeruleus. AB - The effects of acute and short-term administration of tryptophan or tryptophan plus ethanol on serotonin [5-hydroxytryptamine (5-HT)] and two of its metabolites, 5-hydroxyindoleacetic acid (5-HIAA) and 5-hydroxytryptophol (5 HTPL), in the locus coeruleus were investigated in rats by using the microdialysis method. In addition, the acute effects of these drugs on noradrenaline and its metabolite 4-hydroxy-3-methoxymandelic acid (HMMA) were addressed. A single co-administration of tryptophan (50 mg/kg, i.p.) and ethanol (1.25 g/kg, i.p.) did not change the concentrations of either noradrenaline or its metabolite in the locus coeruleus. In contrast, administration of tryptophan (50 mg/kg, i.p.) for three consecutive days caused an increase in the concentration of 5-HIAA, but not that of 5-HT, in the locus coeruleus. Combined administration of tryptophan plus ethanol for 3 days resulted in marked increases in 5-HIAA concentrations in the locus coeruleus, but not in 5-HTPL concentrations. However, administration of ethanol (1.25 g/kg) for 3 days had no effect on the concentrations of 5-HT and its metabolites. The increased 5-HIAA concentration that resulted with combined tryptophan plus ethanol administration was remarkably suppressed by disulfiram. Moreover, in comparison with tryptophan treated rats, the behavioral sign of teeth-chattering was significantly detected in tryptophan plus ethanol-treated rats, but the enhancement of behavioral signs with combined treatment was markedly suppressed by disulfiram. Results of the current study seem to indicate that the stimulation of 5-HT metabolism in locus coeruleus serotonergic neurons by tryptophan was strengthened by the simultaneous administration of ethanol in short-term experiments, and that the increased 5 HIAA concentrations in the locus coeruleus are responsible for behavioral activation. PMID- 15902918 TI - Quantification of silver-staining nucleolar organizer region in epithelial cells of tongue of mice after exposure to, or intake of, alcohol. AB - The purpose of the current study was to evaluate the effect of alcohol on the proliferative activity of epithelial cells in the lingual mucosa of mice by means of silver-staining nucleolar organizer region (AgNOR) count and area measurements. Forty-eight CF1 mice were separated into three groups. The test groups were submitted to topical exposure to, or intake of, 40% (volume/volume) ethyl alcohol. Biopsy specimens were collected from the middle third of the dorsal tongue at 0, 6, and 12 months, and samples were stained according to the AgNOR technique. Mean number and mean area of AgNOR per nucleus were calculated for 50 basal layer cells and 50 intermediate layer cells. Increases in mean number and mean area of AgNOR per nucleus in intermediate cells were observed at 12 months in the alcohol intake group (P < .05). Results showed that intake of 40% alcohol increased epithelial cell proliferation in the dorsal surface of lingual mucosa. PMID- 15902919 TI - Is an alpha-conotoxin MII-sensitive mechanism involved in the neurochemical, stimulatory, and rewarding effects of ethanol? AB - Ethanol and nicotine are the most commonly abused drugs among human beings, and a large body of evidence, from both epidemiologic and preclinical studies, indicates that there is a positive correlation between intake of both drugs. Findings of studies from our research group have demonstrated that nicotinic acetylcholine receptors, especially those located in the ventral tegmental area, are important for the stimulatory, rewarding, and dopamine-enhancing effects of ethanol. Furthermore, results of recent work indicate that the alpha4beta2* and the alpha7* receptor subunits of the nicotinic acetylcholine receptors do not seem to be involved in the neurochemical and behavioral effects of ethanol in rodents. The aim of the current study was to investigate further the role of different nicotinic acetylcholine receptor subunits in the stimulatory, dopamine enhancing, and rewarding effects of ethanol in rodents by using the peptide alpha conotoxin MII (5 nmol; an antagonist of the alpha3beta2*, beta3*, and alpha6* subunits of the nicotinic acetylcholine receptor) administered locally into the ventral tegmental area. A significant reduction of ethanol-induced accumbal dopamine overflow, measured by means of in vivo microdialysis, and of locomotor stimulation was observed in mice. Furthermore, alpha-conotoxin MII was demonstrated to reduce voluntary ethanol intake significantly in both rats and mice. These results indicate that alpha-conotoxin MU-sensitive receptors may be important in mediating the stimulatory, dopamine-enhancing, and rewarding effects of ethanol, and that alpha-conotoxin MII-sensitive receptors may constitute targets for development of new adjuvant for treatment of ethanol dependence. PMID- 15902920 TI - Mice lacking metabotropic glutamate receptor 4 do not show the motor stimulatory effect of ethanol. AB - Group III metabotropic glutamate receptors (mGluRs), specifically receptors 4, 6, 7, and 8 (i.e., mGluR4, mGluR6, mGluR7, mGluR8), play an important role in the generation of locomotion as well as in the behavioral effects of some psychostimulants. Because the arousing or stimulant effects of ethanol seem to be relevant behavioral traits associated with its rewarding properties and genetic susceptibility to alcoholism, we addressed the role of mGluR4 by studying behavioral actions of ethanol in mutant mice lacking mGluR4. Null mutant mice showed higher motor response to novelty than did wild-type mice. Ethanol (1.0-2.5 g/kg) stimulated motor activity of wild-type mice, but not of null mutant mice. There were no significant differences between wild-type and knockout strains in ethanol consumption or preference in two-bottle paradigm, severity of ethanol induced acute withdrawal, or duration of loss of righting reflex. These results show that mGluR4 may play a role in locomotor activity in general and also display specificity for mediation of the motor stimulant effect of ethanol. Consistent with findings of other studies, these results confirm the lack of correlation between ethanol-induced motor stimulation and consumption of ethanol measured in a self-administration paradigm in mice. PMID- 15902921 TI - Accumbal dopamine concentration during operant self-administration of a sucrose or a novel sucrose with ethanol solution. AB - The goal of the current study was to determine the effect of operant self administration of (1) 10% sucrose and (2) a first-time solution of 10% sucrose with 5% or 10% ethanol, on dopamine concentration in the nucleus accumbens. We used an operant procedure that distinguished lever pressing (an appetitive behavior) from drinking to better assess the effect of fluid consumption on accumbal dopamine activity. Male Long-Evans rats were trained to bar press by using 10% sucrose reinforcement, and they were required to emit an escalating number of bar presses across daily sessions. Completion of the response requirement resulted in 20 min of access to the solution. Microdialysis samples were collected before, during, and after bar pressing and drinking, and content of ethanol and dopamine was determined. Dopamine concentration in the dialysate was slightly, but significantly, increased in both groups during lever pressing. However, after consumption began, dopamine concentration increased in the sucrose, but not in the sucrose with ethanol, group, followed by a return to baseline values. Ethanol consumption was low (0.27 +/- 0.02 g/kg) and corresponded to low dialysate ethanol concentrations, which appeared within 5 min of drinking. These results demonstrate that operant self-administration of sucrose increases accumbal dopamine concentration during consummatory phases of behavior, but that a similar increase is not apparent when a novel, perhaps aversive, solution (sucrose with ethanol) is presented. This difference may be due to the sensory-related stimulus properties of each solution. In addition, oral self-administration of ethanol at 0.27 +/- 0.02 g/kg over 20 min is not sufficient for stimulation of dopamine activity in the nucleus accumbens. PMID- 15902922 TI - Relation of tumor necrosis factor (TNF) gene polymorphisms with serum concentrations and in vitro production of TNF-alpha and interleukin-8 in heavy drinkers. AB - Tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) play a role in the pathogenesis of alcoholic hepatitis. The aim of the current study was to investigate the possible relation of TNF gene polymorphisms with TNF-alpha and IL 8 synthesis in heavy drinkers. Nineteen heavy drinkers and 14 healthy control subjects were included in the study. Investigations included (a) polymorphisms in the TNF promoter region at positions -238 (G-->A), -308 (G-->A), -857 (C-->T), and -863 (C-->A), as well as a biallelic Ncol restriction fragment length polymorphism in the first intron of the close TNF-beta gene; (b) serum TNF-alpha and IL-8 concentrations; and (c) TNF-alpha and IL-8 production by phytohemagglutinin A-stimulated peripheral blood mononuclear cells. In comparison with findings for control subjects, heavy drinkers showed higher TNF-alpha production, higher IL-8 production, and higher serum IL-8 concentrations. Increased serum TNF-alpha concentrations were specifically found in heavy drinkers with the -857 (C-->T) substitution (CT heterozygotes), therefore indicating an interaction between alcohol consumption and that polymorphism on serum TNF-alpha concentrations. PMID- 15902923 TI - Effect of a single dose of ethanol on developing skeletal muscle of chick embryos. AB - Fetal alcohol syndrome is a condition occurring in some children of mothers who have consumed alcohol during pregnancy. Many of these affected children show retarded physical growth in the postnatal period despite adequate nutrition. On the basis of findings from studies with animals, it has been proposed that this is due to allometric retardation of growth of skeletal muscle, although the exact reasons for this are not known. The aim of the current study was to examine the structural changes in skeletal muscle in fetal alcohol syndrome in an attempt to understand the mechanisms of growth retardation in fetal alcohol syndrome. Chick embryos were exposed to single doses of 5%, 10%, and 15% ethanol, and the effects on the general growth and development, as well as on the skeletal muscle, of these chicks were studied. There was a significant retardation in crown rump length, head circumference, and body weight in ethanol-exposed chicks when these parameters were compared with findings for appropriate control groups. This retardation was associated with significant and proportionate reductions in the weights of skeletal muscles. Microscopic examination of skeletal muscle showed areas of neutrophil infiltration and necrosis, suggestive of muscle damage, in chicks exposed to 10% and 15% ethanol. Thus, findings of the current study demonstrate the direct toxic effects of a single dose of ethanol on developing embryos in general and skeletal muscle in particular. The pathologic changes seen in skeletal muscle could account for the failure in postnatal growth in fetal alcohol syndrome. PMID- 15902925 TI - Sudden cardiac death caused by innocent chest wall blows. PMID- 15902924 TI - Locomotor and pyretic effects of MDMA-ethanol associations in rats. AB - 3,4-Methylenedioxymethamphetamine [(MDMA) or ecstasy] is a popular club drug often used in combination with ethanol. In the current study, we investigated the effects of MDMA and ethanol combinations on locomotor activity and body temperature of rats. For four consecutive days, male Long-Evans rats were treated daily with a 10-mg/kg dose of MDMA with or without a 1.5-g/kg dose of ethanol. 3,4-Methylenedioxymethamphetamine increased spontaneous activity (on average +1,140%), and this increase was potentiated by ethanol on all days (on average +1,710%). Moreover, ethanol inhibited the MDMA-induced hyperthermia (on average 1.3 degrees C) by the first day of treatment, but not on subsequent treatment days, supporting the suggestion that this effect may undergo tolerance. These observations seem to indicate that combined ethanol-MDMA may induce effects on locomotor activity and thermoregulation that involve separate mechanisms, the first one being less sensitive to tolerance than the second one might be. Results of our study have important implications for understanding the motivation and the health risks of polydrug abusers combining ecstasy and ethanol. PMID- 15902926 TI - Myoblast transplantation for heart failure: where are we heading? PMID- 15902927 TI - Drug-eluting stents. The third revolution in percutaneous coronary intervention. AB - Local stent-based drug delivery (drug-eluting stent - DES) is a new technology aimed to prevent the development of neointimal hyperplasia and restenosis following percutaneous coronary interventions. A number of DESs have been developed using different carrier stents, different kind of coatings, and different drugs. However, to date only two polymer-coated DESs (the Cypher sirolimus-eluting stent from Cordis, Johnson & Johnson, Miami Lake, FL, USA; and the Taxus paclitaxel-eluting stent, Boston Scientific, Natick, MA, USA) have become commercially available after a number of randomized trials showed their ability to reduce late luminal loss, binary restenosis and the need for repeat revascularization when compared to bare metal stents. This review describes the general concept of DES and summarizes the results of the principal clinical trials on DESs, both approved for clinical use or under development. For the marketed stents, we also report the results of the first clinical evaluations in real life and a few insights into the most controversial issues. PMID- 15902928 TI - Multiparametric electrocardiographic evaluation of left ventricular hypertrophy in idiopathic and hypertensive cardiomyopathy. AB - BACKGROUND: Electrophysiological abnormalities underlying the increased arrhythmogenicity of left ventricular hypertrophy (LVH) are still under investigation. The aim of this study was to assess non-invasively the electrophysiologic alterations in two different types of LVH, METHODS: Multiparametric non-invasive ECG analysis (R-R interval, QRS and QT intervals, QT dispersion, T-wave complexity, activation-recovery interval [ARI] dispersion, standard deviation of RR intervals [SDNN], filtered QRS duration [fQRS], root mean-square voltage of the terminal 40 ms of the fQRS [RMS40] and low amplitude signal duration (< 40 microV) in the terminal portion of the fQRS [LAS]) was performed in 57 patients with hypertensive LVH and hypertrophic cardiomyopathy (HCM), and in 105 healthy subjects. RESULTS: The R-R interval and SDNN were similar in hypertrophic patients and controls. QRS and QT intervals were longer in hypertrophic patients without any differences between hypertensive LVH and HCM. QT dispersion, T-wave complexity and fQRS were greater in hypertrophic patients; QT dispersion was the greatest in HCM. ARI dispersion was lesser in hypertrophic patients without any differences between subgroups of LVH. fQRS showed a trend toward higher values in hypertensive patients. LAS at 25 Hz had a trend toward lower values in HCM patients, while LAS at 40 Hz and RMS40 showed no difference between controls and hypertrophic patients. Left ventricular mass index was not correlated with any of the above-mentioned parameters. CONCLUSIONS: The QT interval and dispersion did not identify the type of hypertrophy. Similarly, ARI dispersion which explores local variations of repolarization duration, and T-wave complexity could not distinguish patients with hypertensive LVH from those with HCM indicating that multiparametric ECG data are affected more by the presence of LVH, than by its type. PMID- 15902929 TI - The database of Italian general practitioners allows a reliable determination of the prevalence of myocardial infarction. AB - BACKGROUND: To plan preventive intervention after myocardial infarction (MI) the disease prevalence and the age and time from acute event of the index population should be known. METHODS: We identified all the living patients with MI coded diagnosis in the database of the Italian College of General Practitioners (Health Search Database-HSD). The years from the first acute MI were also determined. RESULTS: 3588 subjects with MI diagnosis were identified (2698 males and 888 females, for 2 gender not recorded). Based on the distribution of our population and on that reported by the Italian Institute of Statistics, stratified by gender and age (segments of 10 years), the estimated number of subjects with MI in Italy (age-standardized rates x 10000) was 309284 for men and 102343 for women. CONCLUSIONS: The prevalence of MI diagnosis in the HSD is very close to that obtained by other epidemiological methods. Querying the database can provide a simple and inexpensive way to estimate and monitor the prevalence of MI in Italy. PMID- 15902930 TI - Fibrinogen as a predictor of mortality after acute myocardial infarction: a forty two-month follow-up study. AB - BACKGROUND: Several studies suggest that fibrinogen may be considered an independent risk factor for coronary artery disease, but it is still on debate if we need its evaluation during an acute myocardial infarction (AMI) to prevent future fatal or non-fatal cardiovascular events. Therefore, we decided to investigate this field. METHODS: We studied 92 male patients with AMI, evaluating at admission age, body mass index, systolic blood pressure, cigarette smoking, ejection fraction, plasma levels of total cholesterol, triglycerides, fibrinogen, glycemia, and white blood cell count. All patients were followed up for 42 months to evaluate total mortality and cardiovascular morbidity. RESULTS: During the follow-up 5 patients died and 64 had one or more non-fatal cardiovascular events: angina (n = 78), heart failure (n = 17), re-AMI (n = 3), stroke (n = 3), or revascularization procedure (n = 16). A multivariate analysis revealed that fibrinogen plasma levels at admission (r = +0.213, p < 0.05) were independently associated with mortality, while systemic thrombolysis was negatively associated (r = -0.447, p < 0.0001). CONCLUSIONS: Plasma fibrinogen levels were the only independent predictor of mortality in a 42-month follow-up post-AMI. This finding, together with other observations from recent studies, suggest that fibrinogen evaluation during AMI may be useful in identifying patients at higher risk of acute event recurrence. PMID- 15902931 TI - Does atrial fibrillation in elderly patients with chronic heart failure limit the efficacy of carvedilol? Suggestions from an observational study. AB - BACKGROUND: No clinical investigation provided any information about a possible influence of atrial fibrillation on the response to beta-blocker therapy in elderly patients with chronic heart failure (CHF). The aim of this study was to observe carvedilol effects in a cohort of patients > 70 years of age with CHF due to left ventricular dysfunction and with chronic atrial fibrillation. METHODS: An observational, 12-month prospective clinical and echocardiographic study was carried out on 240 patients > 70 years of age with heart failure due to systolic dysfunction, 64 of whom with atrial fibrillation. RESULTS: After 1 year of beta blocker treatment, patients with atrial fibrillation and those in sinus rhythm showed similar benefits, in terms of symptomatic improvement (deltaNYHA -0.44 if atrial fibrillation vs -0.57 if sinus rhythm, p = NS), reduction of events (death + hospitalizations -38 vs -15%), recovery of cardiac function (left ventricular ejection fraction delta +8.8 vs +9.4%, p = NS; left ventricular end-diastolic volume delta -17.2 vs -12.5 ml, p = NS), and reduction in mitral regurgitation (delta -042 vs -0.57, p = NS). No difference was found between the two study groups regarding left ventricular end-diastolic volume reduction (12% in atrial fibrillation patients and 18% in sinus rhythm patients, p = NS) and prevalence of the "reverse remodeling" phenomenon (22 and 21%, respectively, p = NS). CONCLUSIONS: In CHF patients > 70 years of age, beta-adrenergic blockade was shown to be equally effective in improving symptoms and left ventricular geometry and function in patients with atrial fibrillation or in sinus rhythm, without any adjunctive sign of long-term clinical deterioration. PMID- 15902932 TI - Dynamic assessment of a composite arterial Y-graft achieving complete myocardial revascularization: transthoracic echo-Doppler correlates with myocardial scintigraphy. AB - BACKGROUND: Composite arterial grafts are presently being used ever more frequently in coronary bypass surgery. We assessed the composite radial artery and in situ left internal thoracic artery Y-graft by means of transthoracic echo Doppler and myocardial perfusion scintigraphy (MPS). METHODS: In 53 patients who underwent complete myocardial revascularization using only this composite arterial graft, good transthoracic echographic images and pulsed Doppler signals of the Y-graft main stem were obtained at rest and early after standard exercise; the echographic parameters were measured. Stress/rest MPS was the gold standard for residual myocardial ischemia. The patients with negative MPS were divided into groups according to the number of coronary vessels grafted and their history of preoperative myocardial infarction. RESULTS: The diastolic-to-systolic ratio of the peak velocities and velocity-time integrals both at rest and after stress, and the stress-to-rest ratio of the diastolic peak velocities and diastolic velocity-time integrals in the negative MPS patients were significantly greater than in the 6 positive MPS patients. However, the restricted number of positive MPS cases limits the power of this statistical analysis. The stress-to-rest ratio of the diastolic velocity-time integrals in the patients with three grafted coronary vessels (median 2.3) and in those without preoperative myocardial infarction (median 2.1) were respectively greater than in the patients with two vessels grafted (median 1.7, p < 0.0001) and those with preoperative myocardial infarction (median 1.8, p = 0.0048). CONCLUSIONS: Dynamic assessment with transthoracic echo-Doppler of a composite arterial graft including the in situ left internal thoracic artery correlates with MPS. In patients with negative MPS, transthoracic echo-Doppler correlates with the myocardial demand. PMID- 15902933 TI - Surgery for acute type A aortic dissection: the effect of antegrade selective cerebral perfusion on the early outcome of elderly patients. AB - BACKGROUND: The aim of this study was to evaluate early results and to determine predictive risk factors associated with an adverse outcome in elderly patients after acute type A aortic dissection repair using antegrade selective cerebral perfusion (ASCP). Adverse outcome was defined as the occurrence of death or permanent neurologic dysfunction. METHODS: From October 1995 to March 2002, 178 patients (group A < 75 years, n = 156, 87.6%; group B > 75 years, n = 22, 12.4%) underwent surgery for acute type A aortic dissection using ASCP and moderate hypothermia. An ascending aorta/hemiarch replacement was performed in 128/178 (71.9%) patients (group A 71.2%, group B 77.3%, p = NS), an ascending aorta and arch replacement in 50/178 (28.1%) patients (group A 28.8%, group B 22.7%, p = NS). Associated procedures were performed in 55/178 (20.9%) patients (group A 31.4%, group B 27.3%, p = NS), the arch vessels were reimplanted using the separated graft technique in 32/50 (64.0%) patients (group A 62.2%, group B 80.0%, p = NS). The mean ASCP time was 59 +/- 27 min. RESULTS: The overall adverse outcome rate was 20.8% (group A 21.2%, group B 18.2%, p = NS). The transient neurologic dysfunction rate was 9.5% (group A 9.5%, group B 5.6%, p = NS). A logistic regression analysis revealed cardiopulmonary bypass time (p = 0.045, odds ratio 1.03/min) to be the only independent predictor of adverse outcome in group A. CONCLUSIONS: During type A aortic dissection repair the implementation of ASCP resulted in an acceptable hospital mortality and neurologic outcome. If ASCP is used, the risk of hospital mortality and postoperative morbidity is similar in patients younger and older than 75 years. Duration of cardiopulmonary bypass still remains an important risk factor for hospital mortality and neurologic outcome in elderly patients. PMID- 15902934 TI - Coronary slow flow phenomenon: description of three cases evaluated with myocardial perfusion scintigraphy. AB - The coronary slow flow phenomenon (CSFP) is an angiographic finding characterized by delayed opacification of the epicardial coronary arteries in the absence of significant stenosis, spasm, dissection or thrombus. Although this poorly understood phenomenon received little attention, patients with CSFP at coronary angiography often suffer from recurrent episodes of chest pain, sometimes occurring during an acute coronary syndrome. We describe 3 cases of patients with CSFP who complained of recurrent chest pain; in one of them an episode of chest pain was so severe as to bring the patient to the emergency department. Indeed, in all our 3 cases myocardial ischemia was evaluated on the basis of a positive myocardial scintigraphy result. In conclusion, it is suggested that CSFP may be an acute and recurrent perturbation of microvascular function with an often severe impairment of quality of life. Myocardial perfusion scintigraphy might help for an accurate assessment of myocardial ischemia in such patients. PMID- 15902935 TI - Unusual case of single coronary artery: questions of methods and basic concepts. AB - Coronary artery anomalies continue to constitute a confusing subject in modern cardiology. While most anomalies are considered to have a benign prognosis, the literature and cardiologic culture frequently imply an intrinsic, systematic association of coronary anomalies with severe clinical presentations. We present a case of unusual single coronary artery, in order to elucidate the logical process that should be used to study similar cases. A 56-year-old female presented with a 6-year history of atypical chest pain and an abnormal electrocardiogram. Heart catheterization revealed an abnormal coronary tree interpreted by some observers as a benign coronary anomaly, by others to indicate the need for coronary angioplasty. A nuclear stress test was performed after 1 year of unrelenting symptoms and showed mildly abnormal findings, leading to a more definitive angiographic study that clarified the anatomy and the prognosis. The case is essentially and only an example of single coronary artery with origin of all branches from the right coronary sinus, but with an unusual triple origin of the branches serving the left anterior descending territory. The notion that a case of single coronary artery may have significant prognostic and clinical repercussion is frequently repeated in the current inconclusive literature. A rational discussion should deal both with individual case objective evidence and theoretical general consideration. PMID- 15902936 TI - Anomalous origin of the left coronary artery from the pulmonary artery in an adult pregnant patient: surgical and percutaneous myocardial revascularization. AB - An anomalous origin of the left coronary artery from the pulmonary artery (Bland White-Garland syndrome) was diagnosed in a 31-year-old woman complaining of angina at 10 weeks of gestation. After termination of pregnancy, the patient underwent surgical repair with ligation of the left coronary artery at the ostium, and a single bypass graft with a left internal thoracic artery to the left anterior descending coronary artery. Angiography, performed at 6 months of follow-up, showed stenosis of the distal anastomosis of the graft that was treated with angioplasty and deployment of a paclitaxel-eluting stent. After 9 months the patient was symptom-free and angiography excluded in-stent restenosis. Experience with the surgical repair of the anomaly is limited by the rarity of this condition; in particular, it has never been reported as a first diagnosis during pregnancy. In the discussion, we analyze the efficacy and limits of the different current strategies for the surgical repair of this rare, but potentially life-threatening, congenital coronary anomaly. PMID- 15902937 TI - An unpleasant surprise in the setting of primary percutaneous coronary intervention: diffuse and severe vessel ectasia with acute thrombosis of the distal right coronary artery in a patient with acute inferior myocardial infarction. AB - Coronary artery ectasia is defined as a > 1.5-fold dilation of the coronary artery compared to the diameter of adjacent normal segments. It must be distinguished from discrete aneurysms that appear in areas adjacent to coronary artery stenosis. It is usually considered a variant of coronary atherosclerosis. Dilated segments are thought to modify the rheology of blood, sluggish or turbulent flow predisposing to myocardial ischemia and its sequelae, including myocardial infarction and sudden death. We report the case of a 52-year-old man, light smoker, with arterial hypertension and family history of coronary artery disease, who was referred to our coronary care unit for an inferior ST-elevation acute myocardial infarction and presented with severe and diffuse vessel ectasia and right coronary thrombosis at coronary angiography. PMID- 15902938 TI - Ischemic stroke in a young woman with aortic papillary fibroelastoma: echocardiographic diagnosis and surgical excision. AB - Papillary fibroelastoma is a benign cardiac tumor typically attached to the cardiac valves. The tumor is recognized during life often in patients evaluated for embolic events of unclear etiology, but sometimes it is recognized in totally asymptomatic patients. We describe the case of a papillary fibroelastoma involving the aortic valve in a 27-year-old woman who presented ischemic stroke. The diagnosis was obtained at two-dimensional echocardiography and confirmed at transesophageal echocardiography. The tumor was located in the non-coronary aortic cusp, without aortic regurgitation. No other embolic source was identified. The tumor was surgically removed. The resection was curative by complete surgical excision, without damage of the aortic valve. The biopsy confirmed the diagnosis. The clinical course was uneventful. This case is an example of a primary intracavitary tumor recognized in a young woman with unclear embolic event. Two-dimensional echocardiography proved to be the exam of choice for the early diagnosis of cardiac tumors. PMID- 15902939 TI - The missing intra-aortic balloon pump catheter. PMID- 15902940 TI - Fluoroscopic guidance is mandatory to insert intra-aortic balloon pump!(?). PMID- 15902941 TI - [How to write a scientific paper]. AB - To write a paper and to succeed in getting it published in a highly renowned scientific journal represent the last but not the least of the difficulties that a researcher has to face before being able to consider a given research project fully accomplished. While writing a manuscript, it is important to put a consistent effort in designing its structure, paying attention to a few technical guidelines. It is worth keeping in mind, however, that the editorial success of a manuscript is related not only to the writer skills and experience but also to the attention previously dedicated to the design and conduction of the research project from which data are derived. On the other hand, the probability for a well designed and carefully performed study (yielding data relevant from a clinical viewpoint), to have its results accepted for publication in an important journal is very low if they are poorly summarized in a low quality paper. In the light of these considerations, the aim of this concise review was to provide the researcher with a few simple tips on how to prepare a scientific article, ranging from its early drafting to its final publication. We have also dealt in some detail with the problems related to the selection of the journal and manuscript format, with the procedures related to manuscript submission, and with the approach to follow in order to prepare a proper answering to the reviewers' and editors' comments. PMID- 15902942 TI - [The electronic mail in the patient-physician relationship]. AB - The continuous spread of e-mail determines an ever-increasing use of this tool for information exchange in healthcare. In spite of that, the frequency and quality of on-line communication between the physician and the patient are still scarce. Beyond analyzing the causes that make difficult the adoption of e-mail in healthcare, and beyond explaining the potential advantages, this paper focus on how this type of communication may influence the patient-physician relationship. More specifically two different types of patient-physician relationship have to be distinguished: type A is characterized by the absence of a preexisting face-to face interaction, whereas type B is characterized by the presence of a preexisting contractual relationship. The management of e-mail messages needs attention and requirements that are different in the two cases. In type A relationship, there apply some principles different from the guidelines prepared by the American Medical Association that refer to type B interaction. The authors describe and comment the principles and guidelines that apply to the two types of patient-physician relationships. The aim of this work was to help physicians to manage and treat e-mail communication with the patient in a suitable way. PMID- 15902943 TI - [Non-ST-elevation myocardial infarction with normal coronary arteries, clinical features and coronary artery flow]. AB - BACKGROUND: No assessment has been made up today concerning clinical features, coronary artery flow and mid-term prognosis between acute non-ST-elevation myocardial infarction (NSTEMI) patients without epicardial coronary disease and those with epicardial coronary artery stenosis > 50% of at least one vessel. METHODS: We evaluated consecutive NSTEMI patients who had undergone coronary angiography within the first 48 hours of infarction. We examined their age, sex, smoking habits, the incidence of diabetes, dyslipidemia, hypertension, and left ventricular ejection fraction. The coronary blood flow was assessed according to the conventional TIMI flow grade and with the TIMI frame count (TFC). RESULTS: From October 1, 2001 to December 31, 2003, 50 patients out of 996 with NSTEMI (20 males, 30 females, mean age 60 +/- 13 years), showed normal coronary arteries (5%). This subset of patients was compared with 50 NSTEMI patients with coronary stenosis. Patients of the first group were younger and more frequently female with respect to NSTEMI patients with coronary stenosis. The differences between the two groups with respect to diabetes, hypertension, dyslipidemia incidence and ejection fraction (52 vs 47%) were not statistically significant. With the corrected TFC (cTFC) method we found a slow flow in at least one coronary vessel in a high percentage of NSTEMI patients with normal coronary arteries. When we compared normal vessels between the two groups, we found a higher cTFC in NSTEMI patients with normal coronary arteries than in NSTEMI patients with coronary stenosis. After a 16 +/- 8 months of follow-up we observed 8 events in the normal vessel group and 10 in the coronary stenosis group (p = NS). CONCLUSIONS: The possible hypothesis of microvessel dysfunction as a pathogenesis of a slow flow in NSTEMI patients with normal coronary arteries is strong. Further studies are warranted to investigate microvessel disease and characteristics and possible causes of abnormalities. A larger perspective study with a longer follow-up is needed as well to evaluate the prognosis in this subset of patients. PMID- 15902944 TI - [Primary angioplasty in a patient with the May-Hegglin anomaly, a rare heredity thrombocytopenia. A case report and review of the literature]. AB - The May-Hegglin anomaly (MHA) is a rare autosomal dominant platelet disorder characterized by thrombocytopenia, giant platelets and leukocyte inclusion bodies. Many patients affected by the MHA have a marked hemorrhagic tendency, a well known contraindication to thrombolytic and anticoagulant therapies. We report a case of a 56-year-old woman with the MHA, referred to our department with an evolving acute ST-elevation myocardial infarction. The patient underwent urgent coronary angiography revealing the acute occlusion of the distal left anterior descending coronary artery, treated with a thrombus aspiration system. In view of the absence of residual stenosis, no balloon dilation and stent deployment were performed. No antiaggregant and anticoagulant therapy was administered. The procedure has been successful, the hospital course was uneventful and the patient was discharged 5 days later. At a 30-day follow-up the patient was asymptomatic and in a good hemodynamic state. To the best of our knowledge, this report is the first description of managing a myocardial infarction in a patient affected by the MHA in the reperfusion era. PMID- 15902945 TI - [Executive summary of the guidelines on the diagnosis and treatment of acute heart failure]. PMID- 15902946 TI - Telling it like it is. PMID- 15902947 TI - Lumbar burst fracture associated with bowel, bladder, and sexual dysfunction: case study. AB - Lumbar burst fractures occur when unusual force and flexion are placed on the spine, causing the vertebral body to rupture and possibly protrude into the spinal canal. A resulting conus medullaris injury is possible, but not common. In this case presentation, a young man suffered bowel, bladder, and sexual dysfunction after a 25-foot fall that caused a burst fracture of the first lumbar vertebra. The presentation's primary focus is the nursing education needed to care for patients who experience the unusual side effect of conus medullaris injury. The psychosocial aspects attributable to age, developmental stage, and stigma for a young man with these dysfunctions also are explored. PMID- 15902948 TI - Spinal epidural abscess-from onset to rehabilitation: case study. AB - Spinal epidural abscess (SEA) is a rare condition; consequently, the clinical significance of early diagnosis often is overlooked. The challenge with SEA is not its treatment, but achieving early diagnosis before neurological symptoms occur. Once neurological deficits are present, immediate treatment must be implemented to prevent deterioration that can result in paralysis. The common SEA signs and symptoms are fever, back pain, and radicular symptoms that can progress to weakness and paralysis. Magnetic resonance imaging is the imaging of choice for diagnosis. Treatments include antibiotics and surgical evacuation of the abscess if neurologic deficits are present. Nurses play a major role in caring for SEA patients by identifying those at risk, preventing complications, and collaborating with a multidisciplinary team to prepare patients for rehabilitation. PMID- 15902949 TI - Health promotion interests of women with disabilities. AB - The number of women with disabilities (WWD) in the United States is substantial and growing. Health promotion education can play a key role in the achievement of optimal health and well-being for this population. A preliminary step in planning educational programs is to identify health promotion topics of interest to this target population. This study surveyed WWD about their interest in health promotion topics and compared their interests by demographic characteristics (i.e., age, employment status, ethnic group, type of disabling condition, and level of disability). The sample population, comprising 604 women with a variety of disabilities, completed a written or online survey. Topic of interest selected were tallied and ranked by order of interest. Topics ranking highest in interest and selected by at least half of the WWD were aging with a disability, stress management, exercise, nutrition, overview of health promotion, and maintaining mental health. Significant health issues not identified included smoking cessation, pregnancy, and osteoporosis. If healthcare providers are aware of the topics that interest WWD, they can better develop educational programs to meet the needs of this vulnerable population. PMID- 15902950 TI - The validity and reliability of the Knowledge of Women's Issues and Epilepsy (KOWIE) Questionnaires I and II. AB - The Knowledge of Women's Issues in Epilepsy (KOWIE) Questionnaires I and II were developed to assess what women with epilepsy (WWE) and practitioners know about relevant topics and concerns. Prior to disseminating any tool, an instrument should be both valid and reliable. The purpose of this study was to report the validity and reliability of the KOWIE Questionnaires I and II. To establish validity, the original KOWIE was sent to five experts who critiqued the relevance of each item. A content validity inventory (CVI) was developed later and sent to 20 additional epilepsy experts across the country. Tool stability was evaluated by test-retest procedures. Patients and practitioners completed corresponding tools on day one, and 24 hours later, on day two. Participants were asked to not review information on the topic of interest until after study procedures were completed. Sixteen of 20 expert responses were included in data analysis; 4 were excluded due to incomplete data. The CVI correlation coefficient was 0.92. Test retest results from all 9 patients and 18 of 20 healthcare professionals were included in data analysis. Correlation coefficients were 0.88 and 0.83 for the KOWIE I and II, respectively, confirming these questionnaires are valid and reliable. While future knowledge may require altering both tools, the current instrument may be used as an assessment tool and guide intervention as it pertains to outcomes in WWE. PMID- 15902951 TI - Participation in a dementia evaluation program: perceptions of family members. AB - Alzheimer's disease (AD) is a chronic, disabling, eventually fatal condition that affects approximately 4 million adults in the United States. The purpose of this qualitative study was to explore several issues from the caregiver's perspective: the experience of having a family member diagnosed with AD, participating in a dementia evaluation program, and complying with that program's recommendations. Interview data from 18 family members revealed (a) a pivotal event led to medical intervention, (b) obtaining the diagnosis was a surprise, and (c) dementia evaluation program recommendations were in place at least marginally 6 months after diagnosis. These caregivers desire assistance from healthcare professionals regarding recommendationsfor appropriate and routine medical and social support and follow-up care. As the population ages and the incidence of AD increases, the perspective of caregivers who make healthcare decisions becomes increasingly important. PMID- 15902952 TI - Health-related quality of life in patients undergoing anterior cervical discectomy fusion. AB - Outcomes are an integral part of surgical decision making among patients, families, physicians, and nurses. The purpose of this descriptive, longitudinal study was to investigate and compare health-related quality of life as an outcome measure in patients undergoing anterior cervical discectomy and fusion (ACDF). Data were collected preoperatively, and at 6 weeks and 3 and 6 months postoperatively. Potential participants were identified by staff members at two neurosurgical practices in central New Jersey. Findings revealed improved health over time with significant change in 6 of the 9 subscales of health-related quality of life. Cervical disk disease and ACDF impact health-related quality of life. A holistic approach to the education and support of patients undergoing ACDF surgery is recommended throughout the preoperative and postoperative periods. PMID- 15902953 TI - The symptoms, neurobiology, and current pharmacological treatment of depression. AB - Depression as a medical disorder increasingly is being recognized and treated. The mood of an individual with major depression is often described as sad, hopeless, or discouraged, and there are many physical symptoms associated with depression. Pharmacologic treatments for depression have advanced greatly since the development of the first therapies, monoamine oxidase inhibitors (MAOIs). Many medications, such as tricyclic antidepressants (TCAs), selective serotonin reuptake inhibitors (SSRIs), and serotonin norepinephrine reuptake inhibitors (SNRIs), currently are available to help combat this health problem. Newer medications have eliminated many of the side effects associated with older therapies, and treatments in development are designed with the goal of further improving on efficacy while eliminating side effects. PMID- 15902954 TI - Stimulation of the subthalamic nucleus for the treatment of Parkinson's disease: postoperative management, programming, and rehabilitation. AB - High-frequency stimulation of the subthalamic nucleus is a neurosurgical procedure for the alleviation of motor symptoms of Parkinson's disease and debilitating medication-induced dyskinesias. Stimulation is achieved with electrodes implanted stereotactically in the subthalamic nucleus by a neurosurgeon specializing in stereotactic surgery and a team composed of an anesthesiologist, a neurophysiologist, certified nurses and nurse practitioners and, at some centers, a neurologist. The teamwork continues in the recovery room and the intensive care unit, where the patient may experience transient adverse behavioral effects. Two weeks after surgery, the neurostimulator is activated and programmed. The medications also are adjusted to complement stimulation to maximize the therapeutic effects and minimize the stimulation-induced side effects. For those patients who are deconditioned or have major speech, gait, or balance problems, rehabilitation therapy is employed. PMID- 15902955 TI - What cues do nurses use to predict aggression in people with acquired brain injury? AB - There is a paucity of research on the frequent and repeated episodes of aggression and violence experienced by nurses when working with people who have an acquired brain injury. The purpose of this study was to bring this issue into focus by identifying the cues nurses use to predict aggression in people with acquired brain injury. Twenty-eight nurses from 10 different inpatient brain injury rehabilitation units in Australia participated in the study. Participants were interviewed using the Critical Decision Method on a one to one basis for up to one and one half hours on two consecutive days. Transcripts of the interviews were analysed using thematic analysis. Results revealed that nurses identified five groups of cues that predict aggression in a patient: (1) what a patient is saying; (2) changes in a patient's voice; (3) changes in a patient's face; (4) changes in a patient's behavior; and (5) a patient's emotions. Nurses reported using multiple cues to predict aggression and highlighted the importance of personal knowledge of the patient in conjunction with identified cues when predicting aggression. Nurses caring for patients with acquired brain injury can predict many episodes of aggression, though not all, by identifying cues from the patient. PMID- 15902956 TI - Discovery of a new biomarker for gastroenterological cancers. AB - Various genomic technologies have been applied to address crucial problems in cancer biology, because cancer develops through the accumulation of various genetic alterations. Of these, gene expression profiling analysis using microarray technology has been widely applied not only to classify cancers at molecular levels, but also to identify novel molecular targets for therapeutics and/or diagnostics. To gain molecular understanding of gastric carcinogenesis, progression, and diversity, we analyzed primary advanced gastric cancer and noncancerous gastric tissues by high-density oligonucleotide microarray. Genes differentially expressed between cancer and noncancerous tissues were identified. In cancer tissues, genes related to cell cycle, growth factor, cell motility, cell adhesion, and matrix remodeling were highly expressed, whereas those related to gastrointestinal-specific function and immune response were rather downregulated. These results provide not only a new molecular basis for understanding biological properties of gastric cancer but also useful resources for future development of therapeutic and diagnostic biomarkers for gastric cancer. Several microarray studies have been published since and have been compared for validation in meta-analysis. As integration of transcriptome information with other biological data is crucial to interpret gene expression data, we have applied oligonucleotide microarray technology to assess allelic gene dosage at 10000 polymorphic loci, namely with an average interval of 200kb. Using a newly developed algorithm, genome imbalance map, loss of heterozygosity (LOH) status can be determined simultaneously. Besides several loci with genomic amplification, we also identified a homozygously deleted chromosomal region in 7q, where frequent chromosomal instability was observed. Finally, we are currently developing novel biomarkers for gastroenterological cancers. Glypican 3 is detected at high levels in serum of hepatocellular carcinoma patients and could be a potential target for antibody therapy. PMID- 15902957 TI - Disease proteomics toward bedside reality. AB - The human genome has been sequenced, and investigation of its products has become possible in a sequence-based framework. More than 200,000 protein species are expressed in the body from approximately 30000 human genes. The term proteome, coined as a linguistic equivalent to the concept of genome, is used to describe the complete set of proteins that is expressed, and modified following expression, by the entire genome in a cell at any one time. Protein types and amounts expressed in a body vary greatly depending upon whether it is healthy or ill. Therefore, proteomics is attracting an increasing interest in its application to better understanding of disease processes, to development of new biomarkers for diagnosis and early detection of disease, and to accelerate drug development. There are numerous opportunities for medicine, although it is quite challenging to meet the needs for high sensitivity and high throughput required for disease-related investigations. PMID- 15902958 TI - Targeting alternatively spliced sequence features for cancer diagnosis and therapeutics. AB - Alternative splicing is emerging as a major new mechanism of functional regulation in mammals, and there is increasing evidence that human cancers often involve significant changes in alternative splicing. In some cases, these changes contribute functionally to the maintenance of the transformed state and could be useful as novel targets for anticancer therapy. In other cases, they reflect changes due to tumorigenesis and could be useful for diagnostic purposes. Fundamentally, alternative splicing offers a novel opportunity to target individual subregions of a gene product that are preferentially expressed in tumors and which are not found in isoforms of the same gene found preferentially in normal tissues. PMID- 15902959 TI - Genotype and phenotype relation in inflammatory bowel disease. AB - Genotype-phenotype analysis helps us to discriminate among various subgroups of IBD and consequently helps us to understand the etiology and pathogenesis as well as develop effective treatments for each subgroup of IBD. PMID- 15902960 TI - Nutritional therapy for Crohn's disease in Japan. AB - In Japan, nutritional therapy as both a primary and as a secondary treatment is widely used for Crohn's disease (CD). The rationale for its use is based on a variety of reasons. The first is its ability to induce remission and to ameliorate the activity of intestinal lesions in the short term by enteral (EN) or by parenteral nutritional therapy in which overexpressions of chemokine receptors in an active stage are decreased significantly in the remission stage. Second is its ability to maintain remission over the long term through home-based enteral nutrition in which tube feeding during the nighttime is encouraged. Third is its ability to reduce the steroid dosage over the period of a long-term treatment course. However, several disadvantages of this therapy such as unpalatability and sluggish effect have been pointed out. Several studies have attempted to resolve this issue and determine the best components of EN, especially in fat composition. Some data have been suggestive of too much long chained fatty acid having a hazardous effect on EN's clinical efficacy because it works as a precursor of inflammatory prostaglandins. Our recent data show that medium-chained triglyceride did not have such a hazardous effect on clinical efficacy. Several studies suggested that the patient factors that were resistant to inducing remission in the short term were a long period of suffering CD, a high activity (on Crohn's Disease Activity Index, CDAI), hemorrhagic colitis, and colitis with marked cobblestoning. Japanese guidelines for the treatment of CD recommended nutritional therapy as a first-line therapy and as a maintenance therapy after inducing remission. This treatment policy has led to Japanese CD patients having lower mortality rates than that of patients who do not receive EN. If this therapy could be combined with other drug therapies, including strong immunosuppressants, treatment strategies would be improved over those we have at present. PMID- 15902961 TI - Treatment of Crohn's disease with anti-IL-6 receptor antibody. AB - We have demonstrated, by using a T-cell transfer murine colitis model, that blocking interleukin 6 (IL-6) signaling with monoclonal antibody (mAb) to IL-6 receptor (IL-6R) abrogated apoptosis resistance of lamina propria T cells, suppressed the expression of vascular adhesion molecules, and successfully prevented and treated intestinal inflammation. Based on these results, we carried out an exploratory clinical trial to investigate the safety and efficacy of humanized anti-IL-6R mAb, MRA, in patients with Crohn's disease (CD). The results were promising, with 80% of the patients given every-2-week MRA infusions for 12 weeks showing a significantly higher clinical response rate as compared to 31% of the placebo-treated patients. Twenty percent of the patients on this regimen went into remission as compared to 0% of the placebo group. The acute-phase responses were normalized by a single MRA infusion, which strongly suggests IL-6 is the major cytokine responsible for their production in CD. PMID- 15902962 TI - Role of strictureplasty in surgical treatment of Crohn's disease. AB - BACKGROUND: The aim of this study was to review early and late results of strictureplasty for patients affected by Crohn's disease. METHODS: We reviewed 103 patients with obstructive Crohn's disease undergoing 293 strictureplasties (Heineke-Mikulicz, 235; Finney, 22; Jaboulay, 35; side-to-side isoperistaltic strictureplasty, 1). Mean age at surgery was 31.4 years. Forty-four patients had at least one previous surgery, and synchronous other surgical procedures were performed in 62 patients. For 41 patients with strictureplasty alone, 154 strictureplasties were done. The site and number of strictures treated by strictureplasty were as follows: duodenum (2), small intestine (265), ileocecal region (6), colon (4), recurrence at previous anastomosis (11), and recurrence at previous strictureplasty (5). The mean number of structureplasties per patient was 2.8. Reoperation has been used as the definitive endpoint for recurrence, and the long-term outcome of strictureplasty was examined. RESULTS: There was no operative mortality. Septic complications related to strictureplasty developed in 4 patients and reoperation was needed in 2 patients (1.9%). Mean duration of follow-up was 80.3 months. For all patients, the 5- and 10-year reoperation rate was 45.0% and 61.9%, respectively. Forty-five patients (43.7%) required further operation for recurrence, of whom 21 patients (20.4%) had recurrence at the site of strictureplasty, which was restricture in 14 patients and perforating disease in 7 patients. Perforating disease for recurrence was more frequent at the site treated by the Finney or Jabouley procedure compared with Heineke-Mikulicz. CONCLUSIONS: It is considered that, in the long term, strictureplasty is safe and useful for preserving the intestine in the surgical treatment of Crohn's disease if strictures are carefully selected. PMID- 15902963 TI - Cisplatin-induced renal interstitial fibrosis in neonatal rats, developing as solitary nephron unit lesions. AB - Cisplatin (CDDP)-induced renal lesions in rats prove a useful model for analysis of the pathogenesis of post-tubular injury-renal interstitial fibrosis. This study investigated the histopathological changes in 10-day-old neonatal rats induced by a single injection of CDDP (4.5 mg/kg). Compared with age-matched controls, on postinjection (PI) days 1 to 6, the number of apoptotic cells, demonstrable with TUNEL method, was significantly increased in CDDP-treated neonates, and there was no marked epithelial necrosis nor fibrotic lesions. Fibrotic lesions began to be developed solitarily around some nephrons with dilated ducts in the corticomedullary junction on PI day 10 and the lesions became more prominent until PI day 20. The alpha-SMA-positive myofibroblastic cells were seen exclusively in the fibrotic lesions. Additionally, the numbers of macrophages reacting with EDI (specific for exudate macrophages), ED2 (for resident macrophages), and OX6 (recognizing MHC class II antigens expressed in antigen-presenting macrophages/dendritic cells) were significantly increased around the affected renal tubules. A greater immunoreaction for TGF-beta1 was seen mostly in the renal epithelial cells of CDDP-treated neonates. These findings indicated that macrophage populations and myofibrolastic cells as well as TGF-beta1 may be responsible for the production of neonatal renal interstitial fibrosis. Compared with CDDP-injected adult rats that develop extensive interstitial fibrosis (Yamate et al., J Comp Pathol, 1995), the formation of fibrotic lesions was delayed, and the lesions were limited to the area around the affected nephrons; this could be attributable to differences in renal morphology between neonates and mature kidney of adult rats. PMID- 15902964 TI - Temporal reduction in size of salivary acinus in rats induced by theophylline. AB - Repeated administration of theophylline, a phosphodiesterase inhibitor, induces the enlargement of the salivary glands in rats. Time-course changes after a single administration of theophylline were examined in the salivary glands, including phosphodiesterase enzyme activity, and the expression of aquaporin 5 (AQP5), a water channel. We also examined the contribution of beta-adrenergic receptors to theophylline-induced salivary changes. Male F344 rats were given 50 mg/kg of theophylline intraperitoneally either alone or concurrently with a 10 mg/kg subcutaneous injection of propranolol. After treatment with theophylline alone, the weight and histology of the submaxillary and parotid glands were examined. Phosphodiesterase activity and AQP5 were detected by enzyme- and immuno histochemistry, respectively. At 4 hours, 8 hours, or both, organ weights were decreased with depletion of secretory vesicles in the acinar cells. In the submaxillary glands, reduced activity of phosphodiesterase and increased expression of AQP5 in the intercalated ducts were observed at 4 hours. When co administered, propranolol partially abolished theophylline-induced glandular reduction. These results suggest that the theophylline-induced transient reduction in size of the salivary glands is attributable not only to phosphodiesterase inhibition but also to beta-adrenergic receptor activation and that the intercalated ducts in submaxillary glands play a role in the production of saliva. PMID- 15902965 TI - Tubular kidney damage and centrilobular liver injury after intratracheal instillation of dimethyl selenide. AB - Accidental inhalation of selenium (Se) derivatives, such as dimethyl selenide (DMSe), has been associated with damage of respiratory tissues. However, systemic effects of inhaled Se have not been thoroughly established. We have investigated whether mouse kidney and liver show cellular pathology as a result of a single intratracheal instillation of two different doses of DMSe (0.05 and 0.1 mg Se/kg BW). The animals were sacrificed 1, 7, 14, and 28 days after either 1 of the 2 DMSe treatments; samples were studied by light microscopy. Instillation of the low DMSe dose resulted in acute and transient tubular disease of the kidney expressed by swelling and vacuolation of epithelial cells of proximal tubules; in some mice, tubular necrosis was observed. After 14 days of the DMSe treatment, these lesions were ameliorated and, by day 28, the kidney tubular epithelium depicted a normal morphology. The same low dose of DMSe caused sustained damage to centrilobular hepatocytes characterized by swollen and vacuolized liver cells. After the instillation of the high DMSe dose, the mice presented sustained liver and kidney focal necrosis. Our data suggest that inhalation of DMSe results in: (i) acute tubular injury of the kidney and damage to centrilobular liver cells and (ii) this systemic pathology induced by DMSe is a dose-dependent phenomenon. PMID- 15902966 TI - Histopathological study of time course changes in PTHrP-induced incisor lesions of rats. AB - Parathyroid hormone related peptide (PTHrP) was discovered as a causative factor of humoral hypercalcemia of malignancy (HHM). In the present study using HHM model rats, the time course of odontoblastic response to PTHrP and its relation to incisal fracture were elicited. Nude rats were implanted with PTHrP-expressing tumor (LC-6) cells, mandibular incisors were collected at several time points. Microscopically 3 distinctive types of odontoblastic/dentin lesions were observed. Hypercalcfied dentin, which was reported as hypercalcemia-induced lesion in previous reports, observed in all areas of the dentin from week 5-10 samplings. Dentin niche, observed solely in week-10 sampling point, exhibited a nature identical to that of reparative odontoblast reported in the literatures of various cytotoxic agents. Since cytotoxicites were neither observed prior to the lesions nor reported as a role of PTHrP, the reparative response may have derived from highly sustained levels of PTHrP. Loss of columnar odontoblasts height was initially observed at week-5 time point in the middle section of the incisor. This primary loss of cell height prior to incisor fracture was considered to be the earliest response to the increased PTHrP levels of this model. PMID- 15902967 TI - Differential expression of COX-1 and COX-2 in the gastrointestinal tract of the rat. AB - The aim of this study was to use immunohistochemistry with morphometry to investigate COX-1 and COX-2 expression in the normal rat gastrointestinal (GI) tract and examine if sites of ulceration previously observed with long-term COX-2 inhibitor administration in mice correlate with differential COX-1/COX-2 expression. COX-2 positive cells were observed predominantly in the apical lamina propria of intestinal villi with fewer cells in the mucosal epithelium. The highest level of COX-2 expression was observed at the ileocaecal junction (ICJ). COX-2 expression was also present in parasympathetic ganglia of the submucosa and muscularis. In the stomach, the highest grade of COX-2 expression was observed in the apical lamina propria of the fundus adjacent to the junctional ridge. In contrast, COX-1 positive cells within the lamina propria were evenly distributed along the GI tract but were present in higher numbers than COX-2 positive cells. The mean level of COX-1 expression at the ICJ was not significantly different from the ileum and caecum. Evidence that the highest level of COX-2 expression in normal rats is located on the ileal side of the ICJ provides the first mechanism to explain spontaneous ulceration and perforation of the distal ileum in COX-2 -/ animals. PMID- 15902968 TI - Statin-induced muscle necrosis in the rat: distribution, development, and fibre selectivity. AB - Simvastatin and cerivastatin have been used to investigate the development of statin-induced muscle necrosis in the rat. This was similar for both statins and was treatment-duration dependent, only occurring after 10 days had elapsed even if the dose was increased, and still occurring after this time when dosing was terminated earlier as a result of morbidity. It was then widespread and affected all areas of the muscular system. However, even when myotoxicity was severe, particular individual muscles and some types of fibres within affected muscles were spared consistently. Fibre typing of spared muscles and of acutely necrotic fibres within affected muscles indicated a differential fibre sensitivity to statin-induced muscle necrosis. The fibres showed a necrotic response to statin administration that matched their oxidative/glycolytic metabolic nature: Least sensitive --> I < - > IIA < - > IID < - > IIB <-- most sensitive. Type I and IIB fibres represent metabolic extremes of a continuum of metabolic properties through the fibre types with type I fibres most oxidative in metabolism and type IIB fibres most glycolytic. In addition, in some (nonnecrotic) glycolytic fibres from muscles showing early multifocal single fibre necrosis the only subcellular alterations present in isolation of any other changes were mitochondrial. These changes were characterised by an increased incidence of vacuolation and the formation of myelinoid vesicular bodies that accumulated in the subsarcolemmal areas. These findings suggest an important early involvement of mitochondria in selective glycolytic muscle fibre necrosis following inhibition of the enzyme HMG CoA reductase. PMID- 15902969 TI - Comparative in vivo toxicity of topical JP-8 jet fuel and its individual hydrocarbon components: identification of tridecane and tetradecane as key constituents responsible for dermal irritation. AB - Despite widespread exposure to military jet fuels, there remains a knowledge gap concerning the actual toxic entities responsible for irritation observed after topical fuel exposure. The present studies with individual hydrocarbon (HC) constituents of JP-8 jet fuel shed light on this issue. To mimic occupational scenarios, JP-8, 8 aliphatic HC (nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane) and 6 aromatic HC (ethyl benzene, o-xylene, trimethyl benzene, cyclohexyl benzene, naphthalene, dimethyl naphthalene) soaked cotton fabrics were topically exposed to pigs for 1 day and with repeated daily exposures for 4 days. Erythema, epidermal thickness, and epidermal cell layers were quantitated. No erythema was noted in 1-day in vivo HC exposures but significant erythema was observed in 4-day tridecane, tetradecane, pentadecane, and JP-8 exposed sites. The aromatic HCs did not produce any macroscopic lesions in 1 or 4 days of in vivo exposures. Morphological observations revealed slight intercellular and intracellular epidermal edema in 4-day exposures with the aliphatic HCs. Epidermal thickness and number of cell layers significantly increased (p < 0.05) in tridecane, tetradecane, pentadecane, and JP-8-treated sites. No significant differences were observed in the aromatic HC-exposed sites. Subcorneal microabscesses containing inflammatory cells were observed with most of the long-chain aliphatic HCs and JP-8 in 4-day exposures. Ultrastructural studies depicted that jet fuel HC-induced cleft formation within intercellular lipid lamellar bilayers of the stratum corneum. The degree of damage to the skin was proportional to the length of in vivo HC exposures. These data coupled with absorption and toxicity studies of jet fuel HC revealed that specific HCs (tridecane and tetradecane) might be the key constituents responsible for jet fuel-induced skin irritation. PMID- 15902970 TI - Susceptibilities of p53 knockout and rasH2 transgenic mice to urethane-induced lung carcinogenesis are inherited from their original strains. AB - In the present study, susceptibility of CB6F1 mice carrying the human prototype c Ha-ras gene (rasH2 mice) and p53 gene knockout mice (p53 (+/-) mice) to urethane induced lung carcinogenesis was compared under the same experimental conditions. Both strains were administered 500 ppm urethane in their drinking water for 3 weeks. At week 26, lung adenocarcinomas and adenomas were observed in 53% and 100% of rasH2 mice, respectively, and lung adenomas were observed in 67% of rasH2 littermate (non-Tg) mice. However, lung tumors were not observed in either p53 (+/-) or p53 (+/+) mice. Peliosis hepatis and hepatic hemangiomas were observed in 27% and 67% of p53 (+/-) mice, but only in 6.7% and 6.7% of the rasH2 animals, respectively. Under the same experimental conditions, BALB/c mice, the strain of origin of the rasH2 mice, developed lung adenomas at an incidence of 93%, whereas none of the C57BL/6 original strain for p53 (+/-) mice developed lung tumors. Peliosis hepatis was observed in 40% of the C57BL/6 mice, but not in BALB/c mice; hepatic and splenic hemangiomas were not observed in these animals. These results indicate that organ susceptibility of rasH2 and p53 (+/-) mice is inherited from their strains of origin, the rasH2 and BALB/c lines being much more sensitive to the induction of pulmonary carcinogenesis. PMID- 15902971 TI - 2-Butoxyethanol female-rat model of hemolysis and disseminated thrombosis: X-ray characterization of osteonecrosis and growth-plate suppression. AB - We recently proposed a chemically induced rat model for human hemolytic disorders associated with thrombosis. The objective of the present investigation was to apply a noninvasive, high-magnification X-ray analysis, the Faxitron radiography system, to characterize the protracted bone damage associated with this 2 butoxyethanol model and to validate it by histopathology. Groups of female Fischer 344 rats were given 0, 250, or 300 mg of 2-butoxyethanol/kg body weight daily for 4 consecutive days. Groups were then sacrificed 2 hours or 26 days after the final treatment. The treated animals displayed a darkened purple-red discoloration on the distal tail. Histopathological evaluation, including phosphotungstic acid-hematoxylin staining of animals sacrificed 2 hours after the final treatment, revealed disseminated thrombosis and infarction in multiple organs, including bones. The Faxitron MX-20 specimen radiography system was used to image selected bones of rats sacrificed 26 days posttreatment. Premature thinning of the growth plate occurred in the calcaneus, lumbar and coccygeal vertebrae, femur, and ilium of the treated animals. Areas of decreased radiographic densities were seen in the diaphysis of the femur of all treated animals. The bones were then examined histologically and showed a range of changes, including loss or damage to growth plates and necrosis of cortical bone. No thrombi were seen in the animals sacrificed at 30 days, but bone and growth plate changes consistent with prior ischemia were noted. The Faxitron proved to be an excellent noninvasive tool that can be used in future studies with this animal model to examine treatment modalities for the chronic effects of human thrombotic disorders. PMID- 15902972 TI - Carcinogenicity evaluation: comparison of tumor data from dual control groups in the Sprague-Dawley rat. AB - Following recent clarification in Europe that a single control group is now acceptable for rodent carcinogenicity studies, the use of dual controls may be reduced or disappear. To date, virtually nothing has been published on whether this latter situation has improved the identification of tumorigenic risk potential in these studies. In this paper, the results of 13 rat carcinogenicity studies, performed between 1991 and 2002, with 2 control groups, are presented. Although no major differences in tumor incidences between these dual control groups were found, some interstudy variation occurred. In cases where a notable difference was seen, the use of 2 control groups, as well as robust, contemporary background data, allowed an easier interpretation of findings in drug-treated groups. Thus, the continued use of dual control groups has a vital role in the assessment of tumoriogenic risk. The paper also presents an update on survival, on the range and extent of background spontaneous neoplasms, and comments on genetic drift in this commonly used rat strain. PMID- 15902973 TI - Analysis of p53 tumor suppressor gene, H-ras protooncogene and proliferating cell nuclear antigen (PCNA) in squamous cell carcinomas of HRA/Skh mice following exposure to 8-methoxypsoralen (8-MOP) and UVA radiation (PUVA therapy). AB - Treatment with 8-methoxypsoralen (8-MOP) and ultraviolet radiation (primarily UVA), called PUVA therapy, has been used to treat different chronic skin diseases but led to a significant increased risk for skin cancer. The National Toxicology Program (NTP) performed a study in mice treated with PUVA that showed a significant increase in squamous cell carcinomas of the skin. In the present study, we evaluated the protein expression of p53 and PCNA and DNA mutations of p53 and H-ras genes in both hyperplastic and neoplastic squamous cell lesions from the NTP study. By immunohistochemical staining, protein expression of both p53 and PCNA was detected in 3/16 (19%) of hyperplastic lesions and 14/17 (82%) of SCCs in groups treated with both 8-MOP and UVA. The mutation frequency of p53 in SCCs from mice administered 8-MOP plus UVA was 15/17 (88%) with a predominant distribution of mutations in exon 6 (14/15 - 93%). No H-ras mutations were detected in the hyperplastic lesions/tumors. The mutagenic effect of PUVA on the p53 tumor suppressor gene may lead to a conformational modification and inactivation of the p53 protein, which are considered critical steps in PUVA induced skin carcinogenesis. The p53 mutational frequency and patterns from our study were different from those reported in human PUVA-type tumors. PMID- 15902974 TI - Morphology of the fetal rat testis preserved in different fixatives. AB - Histopathological examination of the testes of exposed fetuses and neonates is important in assessing the developmental effects of environmental toxins, including sex hormone modulators. Modified Davidson's fluid (mDF) has been suggested as a superior substitute for Bouin's fluid for fixation of adult animal testes. We compared the morphology of fetal rat testes stained with hematoxylin and eosin (H&E) or immunochemically after fixation in 10% neutral buffered formalin (NBF), Bouin's fluid, or mDF. Fixation in mDF resulted in more sharply defined nuclear detail and better preservation of cellular cytoplasm on H&E stained sections of rat testes on gestation day 19. Use of Bouin's fluid did not allow satisfactory detection of apoptotic cells by fluorescent terminal deoxynucleotide transferase-mediated deoxy-UTP nick labeling. Staining with the immunoperoxidase system and the conventional chromogen diaminobenzidine tetrahydrochloride to visualize 5-bromo-2-deoxyuridine-positive cells demonstrated that the number of positive nuclei and intensity of staining were similar with all 3 fixatives. Immunostaining for cytoskeletal protein vimentin was more intense and provided better details of the Sertoli cell cytoplasm with formalin fixation than with mDF. Our study demonstrates that fixation in mDF provided better morphologic detail in the fetal rat testis compared with 10% NBF and Bouin's fluid and illustrates the importance of establishing the correct fixation conditions for each immunostaining protocol. PMID- 15902975 TI - Measurement of trihalomethanes and methyl tert-butyl ether in whole blood using gas chromatography with high-resolution mass spectrometry. AB - The prevalence of disinfection by-products in drinking water supplies has raised concerns about possible adverse health effects from chronic exposure to these compounds. To support studies exploring the relation between exposure to trihalomethanes (THMs) and adverse health effects, an automated analytical method was developed using capillary gas chromatography (GC) and high-resolution mass spectrometry (MS) with selected ion mass detection and isotope-dilution techniques. This method quantified trace levels of THMs (including chloroform, bromodichloromethane, dibromochloromethane, and bromoform) and methyl tert-butyl ether (MTBE) in human blood. Analyte responses were adequate for measuring background levels after extraction of these volatile organic compounds with either purge-and-trap extraction or headspace solid-phase microextraction (SPME). The SPME method was chosen because of its ease of use and higher throughput. Detection limits for the SPME GC-MS method ranged from 0.3 to 2.4 ng/L, with linear ranges of three orders of magnitude. This method proved adequate for measuring the THMs and MTBE in most blood samples tested from a diverse U.S. reference population. PMID- 15902976 TI - Dioxins and furans determination in postmortem blood by gas chromatography--high resolution mass spectrometry. AB - Dioxins and related compounds (furans) are persistent environmental contaminants that cause adverse biological effects. Their influence on humans is still unclear, except for accidental high-dose exposure. However, chronic exposure to these compounds seems to be involved in cancer, endocrine disruption, and neurobehavioral effects. For several years, a large concern about the potential health risks of dioxins is emerging in Europe and United States. Dioxin levels in biological specimens are extremely low and require very sensitive and specific methods of analysis. In this study, gas chromatography coupled to high-resolution mass spectrometry was used to evaluate dioxin body burden of two women deceased from generalized cancer. Fat fraction of blood specimens was obtained after precipitation with ethanol and extraction of both liquid and solid phases spiked with labeled 13C12-dioxin analogues. Organic phases were grouped, washed, and evaporated to weigh the lipid content. Lipids were dissolved in hexane, hydrolyzed with concentrated sulfuric acid, and discarded during water washes. Dioxins purification was achieved using three successive columns: silica, alumina/sodium sulfate, and carbon/Celite. Finally, the toluene eluent was evaporated and the extract injected in the analytical system. After chromatographic separation, detection was achieved in single ion monitoring mode using a high-resolution mass spectrometer operating in electron impact ionization mode (40 eV, minimal resolution of 10,000). Dioxin levels were expressed in pg TEQ/g of fat as defined by the World Health Organization. Quantification limits for each dioxin congener ranged from 2.5 to 12.0 pg/g fat with a relative extraction recovery always higher than 60%. Dioxin concentrations in the blood of the two deceased women were 35.0 and 42.7 pg TEQ/g fat, respectively. These concentrations are largely lower than those observed after accidental releases, but in the range of those observed in the general European population. Therefore, it was not possible to correlate dioxin body burden of the two women as a potential contributor of their cancer pathology. Nevertheless, knowledge of dioxin body burden in the French population would be of interest for an accurate interpretation of these results. PMID- 15902977 TI - Multi-residue determination of anti-inflammatory analgesics in sera by liquid chromatography--mass spectrometry. AB - Non-steroidal anti-inflammatories (NSAIDs) are analgesic, antipyretic, and, as their name implies, anti-inflammatory drugs, which are widely used for the treatment of a variety of human and veterinary disease conditions in which control of pain and inflammation is desired. Acetaminophen (ACE) is a common over the-counter analgesic. Detection of a variety of widely used NSAIDs and ACE in fluid and tissue samples is an important diagnostic tool. A sensitive and selective analytical method has been developed for simultaneous screening of 12 NSAIDs and ACE by liquid chromatography-mass spectrometry with an atmospheric pressure chemical ionization interface set to operate in the negative ion mode of MS. Following sample preparation, all analytes were separated on a C18-reversed phase column with a gradient elution of acetonitrile and acetic acid. Full-scan mass spectral fragmentation profiles were established for each analyte and individual extracted ion chromatograms were used for quantitation. Linearity of detection was observed over the 0.05-25.0 microg/mL range of standard concentrations. The instrument limits of detection (LOD), based on an individual analyte quantitation ions, fell between 0.05 and 1.0 microg/mL for all compounds. The matrix LODs were determined to be 0.05 microg/mL for phenylbutazone (m/z 307); 0.1 microg/mL for indomethacin (m/z 312), flunixin (m/z 295), and piroxicam (m/z 330); 0.5 microg/mL for ACE (m/z 150), diclofenac (m/z 250), ketoprofen (m/z 209), and mefenamic acid (m/z 240); 1.0 microg/mL for oxyphenbutazone (m/z 323); 5.0 microg/mL for ibuprofen (m/z 205), salicylic acid (m/z 137), and tolmetin (m/z 212); and 10 microg/mL for naproxen (m/z 185). PMID- 15902978 TI - Determination of acetonitrile and cyanide in rat blood: application to an experimental study. AB - Methods were developed for the analysis of acetonitrile and its metabolite cyanide in the blood of rats exposed to acetonitrile. Acetonitrile was analyzed by the headspace technique coupled to gas chromatography with detection by flame ionization, and cyanide was analyzed by high-performance liquid chromatography with fluorescence detection (lambdaex = 418 nm and lambdaem = 460 nm) after derivatization of the ion with naphthalene 2,3-dicarboxyaldehyde and taurine. The quantitation limits of the methods for the analysis of acetonitrile and cyanide were 4.875 microg/mL and 0.025 microg/mL, respectively. The coefficients of variation of 10% or less obtained for intra- and interassay precision indicate the precision of these analytical methods and the systematic errors, all less than 5%, indicate that the methods are quite accurate. The methods were applied to an experimental study after the animals received acetonitrile at the doses of 2 mmol/kg or 5 mmol/kg. PMID- 15902979 TI - Simultaneous congener-specific determination of selected organochlorine compounds and nitro musks in human whole blood samples by solid-phase extraction and capillary gas chromatography with electron capture detection. AB - Chlororganic compounds like pesticides or polychlorinated biphenyls (PCB) and nitro musks are environmental contaminants, which remain public health concerns because of their persistence in humans and their toxicological properties. Many of these substances are associated with endocrine dysfunction or with carcinogenicity. Therefore, a simple method using solid-phase extraction followed by capillary gas chromatography with electron capture detection for the simultaneous determination of both organochlorines and nitro musks in human whole blood samples has been developed. Recovery rates of 13 PCB congeners and of 7 pesticides ranged from 67.5% to 100.4% and from 81.1% to 110.5%, respectively. Recoveries of the 5 nitro musks were consistent and ranged from 90.2% to 98.8%. The accuracy for organochlorines and nitro musks varied from 6.3% to 8.6%. Method detection limits ranged from 0.02 microg/L to 0.11 microg/L for the organochlorines and from 0.04 microg/L to 0.08 microg/L for the nitro musks. The method has a high sensitivity with a low detection limit even in slightly contaminated human blood samples. The time and technical effort is small, so the method is feasible for epidemiological studies with regard to the impact of organochlorines and nitro musks on certain diseases. PMID- 15902980 TI - Urinary excretion of alpha-hydroxytriazolam following a single dose of halcion. AB - As an approved medicinal product and reportedly an abused substance that have been associated with death and "considered to be a factor...of impaired driving, sexual assault, and other violent crimes", triazolam is controlled at the same level (Level III) as flunitrazepam in Taiwan. Alleged misuses of this substance have been associated with case specimens submitted to this laboratory. A sample preparation (with and without enzymatic hydrolysis) and gas chromatography-mass spectrometry protocols were evaluated and applied to the analysis of free and total alpha-hydroxytriazolam (the main metabolite of triazolam) in urine. Ions designated for TMS-derivatized alpha-hydroxytriazolam and alpha-hydroxytriazolam d4 are m/z 415, 417, and 430 and 419, 421, and 434, respectively. The overall protocol achieved the following results when applied to the analysis of 2-mL drug free urine specimens fortified with 10-200 ng/mL alpha-hydroxytriazolam: recovery, 95%; interday and intraday precision ranges, 1.50-3.52% and 0.93-4.71%, respectively; linearity, r2 > 0.99; and limits of detection and quantitation, 0.05 and 0.1 ng/mL, respectively. This protocol was applied to the analysis of case specimens and urine samples collected from two patients (A and B) taking one oral dose of Halcion (0.25 mg triazolam). Excretion profiles of free and total alpha-hydroxytriazolam show that free alpha-hydroxytriazolam is detectable, but at very low levels (< 5 ng/mL). Peak excretion of total alpha-hydroxytriazolam occurs at approximately 5-10 h following the drug intake. Total alpha hydroxytriazolam is excreted at detectable levels approximately 2-35 h following an oral dose of 0.25 mg triazolam. Total free and conjugated alpha hydroxytriazolam excreted by A and B are 0.61% and 31.6%; and 0.36% and 57.2% of the dose, respectively. PMID- 15902981 TI - Quantification and profiling of 19-norandrosterone and 19-noretiocholanolone in human urine after consumption of a nutritional supplement and norsteroids. AB - Nandrolone is one of the synthetic anabolic steroids banned in sports and has been a popular substance abused by athletes in recent years. One of its major metabolites, 19-norandrosterone (19-NA), has been used as a determinant for drug violations in sports. Current reports regarding nandrolone-positive cases have been related to intake of some nandrolone-free nutritional supplements. The aim of this study was to learn whether if a nutritional supplement sold by over-the counter (OTC) nutritional stores could yield the same metabolic products as that of nandrolone. If so, what is (are) the substance(s) that contributed to the nandrolone metabolites? To determine the content of an OTC nutritional supplement, a tablet was dissolved in methanol, followed by N-methyl-N trimethylsilyltrifluoroacetamide (MSTFA)-trimethyliodosilane (TMIS) derivatization prior to gas chromatography-mass spectrometry (GC-MS) analysis. The collected urine samples underwent extraction, enzymatic hydrolysis, and derivatization before the analyses of GC-MS. The results showed that seven anabolic steroids were found as contaminants in the nutritional supplement, in addition to six that were listed in the ingredients by the manufacturer. We confirmed previous reports that administration of the OTC supplement could produce a positive urine test for nandrolone metabolites. Furthermore, the results from excretion studies showed that 19-NA and 19-noretiocholanolone (19 NE) were present in urine after consuming the nutritional supplement, nandrolone, 19-nor-4-androsten-3,17-dione, 19-nor-4-androsten-3beta,17beta-diol, and 19-nor-5 androsten-3beta,17beta-diol. The 19-NA concentrations in urine were generally higher than that of 19-NE (19-NA/19-NE ratio > 1.0) especially during the early stage of excretion, that is, before 6 h post-administration. After this period of time, the concentrations of 19-NA and 19-NE fluctuated and might even have reversed (19-NA/19-NE ratio < 1.0) in their ratio, that is, higher yield in 19-NE than that in 19-NA. On the basis of this study, we postulate that some doping violations of nandrolone could be attributed by indiscriminate administration of the OTC nutritional supplements that contained 19-norsteroids. PMID- 15902982 TI - A new GC-MS method for the determination of five amphetamines in human hair. AB - A new gas chromatography-mass spectrometry method for the simultaneous identification and quantitation of amphetamine (AP), methamphetamine (MA), 3,4 methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethamphetamine (MDEA) in hair is proposed. Hair was hydrolyzed in 1 M NaOH at 40 degrees C, subjected to extraction with 4:1 (v/v) methylene chloride/isopropanol, and derivatized with pentafluoropropionic anhydride (PFPA) and ethyl acetate. Calibration curves for the five analytes were constructed over the concentration range 0.5-25.0 ng/mg, using their pentadeuterated analogues as internal standards. The limits of detection and quantitation obtained were 0.045 and 0.151 ng/mg for AP; 0.014 and 0.048 ng/mg for MA; 0.013 and 0.043 ng/mg for MDA; 0.017 and 0.057 ng/mg for MDMA; and 0.007 and 0.023 ng/mg for MDEA. The accuracy of the method was found to be in the range +/- 9%, and the coefficients of variation were less than 8%. Overall, 24 hair specimens tested positive for one or more amphetamines, with average concentrations of 0.88 ng/mg for AP, 10.14 ng/mg for MA, 1.30 ng/mg for MDA, and 8.87 ng/mg for MDMA. Only one specimen tested positive for MDEA with a concentration of 0.84 ng/mg. PMID- 15902983 TI - Pharmacokinetics of the active metabolites of ethyl loflazepate in elderly patients who died of asphyxia associated with benzodiazepine-related toxicity. AB - We determined the pharmacokinetics of ethyl loflazepate (Lof) in elderly patients who died of benzodiazepine-related toxicity. Three elderly patients with body mass indexes of less than 17 kg/m2 died of asphyxia after having taken maintenance doses of Lof for 2 to 3 weeks. We measured serum concentrations of the active metabolites of Lof using gas chromatography-mass spectrometry and a benzodiazepine receptor assay to determine the pharmacokinetics of each. On admission, the serum concentrations of the active metabolites of Lof ([Lofl) were 256 ng/mL, 425 ng/mL, and 177 ng/mL in cases 1, 2, and 3, respectively. Serum benzodiazepine-receptor binding activities, expressed as diazepam equivalent concentrations ([Bz]), were 1800 ng/mL, 2200 ng/mL, and 1500 ng/mL. The T1/2(beta) of [Lof] were 124 and 121 h in cases 1 and 2 and the T1/2(beta) of [Bz] were 75 and 87 h. The distribution volume in the elderly was reduced due to a small lipid compartment, and total drug clearance was decreased due to the decline in liver and kidney function. These changes did not prolong T1/2(beta) but did increase plasma concentrations of active metabolites, especially in case 2, and a slight decrease in protein binding increased the amount of free active metabolites greatly. PMID- 15902984 TI - Plasma-to-blood ratios of congener analytes. PMID- 15902985 TI - Effect of high-fat or high-carbohydrate diets on endurance exercise: a meta analysis. AB - The purpose of this investigation was to clarify, via a meta-analysis, whether the literature favors a high-fat or a high-carbohydrate diet to yield superior endurance exercise performance. Twenty published trials were analyzed to compare exercise performance under different diets. The average effect size of -0.60 indicated that subjects following a high-carbohydrate diet exercised longer until exhaustion. The training status of subjects (trained vs. untrained) was significantly related to effect size (r = -0.576, P < 0.01) and effect sizes separated between trained and untrained subjects were -0.05 and -2.84 respectively. The test for homogeneity revealed significant heterogeneity among effect sizes (chi2 [19] = 43.30, P < 0.05), indicating all of the trials are not describing the same effect. Given this significant heterogeneity, a conclusive endorsement of a high-carbohydrate diet based on the literature is difficult to make. Highly dissimilar trial protocols are the primary reason for heterogeneity. PMID- 15902986 TI - The effects of medium-chain triacylglycerol and carbohydrate ingestion on ultra endurance exercise performance. AB - The aims of the study were to determine if medium-chain triacylglycerol (MCT), ingested in combination with carbohydrate (CHO), would alter substrate metabolism and improve simulated competitive ultra-endurance cycling performance. Eight endurance-trained cyclists took part in this randomized, single-blind crossover study. On two separate occasions, subjects cycled for 270 min at 50% of peak power output, interspersed with four 75 kJ sprints at 60 min intervals, followed immediately by a 200 kJ time-trial. One hour prior to the exercise trials, subjects ingested either 75 g of CHO or 32 g of MCT, and then ingested 200 mL of a 10% CHO (wt/vol) solution or a 4.3% MCT + 10% CHO (wt/vol) solution every 20 min during the CHO and MCT trials, respectively. During the constant-load phases of the 270 min exercise trial, VO2. RER, and heart rate were measured at 30 min intervals and gastrointestinal (GI) symptoms were recorded. There was no difference in VO2 or RER between the MCT and CHO trials (P = 0.40). Hourly sprint (P = 0.03 for trial x time interaction) and time-trial times (14:30 +/- 0.58 vs. 12:36 +/- 1:6, respectively, P < 0.001) were slower in the MCT than the CHO trial. Half the subjects experienced GI symptoms with MCT ingestion. In conclusion, MCTs ingested prior to exercise and co-ingested with CHO during exercise did not alter substrate metabolism and significantly compromised sprint performance during prolonged ultra-endurance cycling exercise. PMID- 15902987 TI - Excess post-exercise oxygen consumption following continuous and interval cycling exercise. AB - The purpose of this investigation was to examine the effect of interval (INT) and continuous (CON) cycle exercise on excess post-exercise oxygen consumption (EPOC). Twelve males first completed a graded exercise test for VO2max and then the two exercise challenges in random order on separate days approximately 1 wk apart. The INT challenge consisted of seven 2 min work intervals at 90% VO2max, each followed by 3 min of relief at 30% VO2max. The CON exercise consisted of 30 to 32 min of continuous cycling at 65% VO2max. Gas exchange and heart rate (HR) were measured for 30 min before, during, and for 2 h post-exercise. Three methods were used to analyze post-exercise oxygen consumption and all produced similar results. There were no significant differences in either the magnitude or duration of EPOC between the CON and INT protocols. HR, however, was higher (P < 0.05) while respiratory exchange ratio (RER) was lower (P < 0.05) following INT. These results indicate that when total work was similar, the magnitude and duration of EPOC were similar following CON or INT exercise. The differences in HR and RER during recovery suggest differential physiological responses to the exercise challenges. PMID- 15902988 TI - Increased caloric intake soon after exercise in cold water. AB - We examined the acute effect of cold-water temperature on post-exercise energy intake (EI) for 1 h. In a randomized, crossover design, 11 men (25.6 +/- 5 y) exercised for 45 min on a submersed cycle ergometer at 60 +/- 2% VO2max in 33 degrees C (neutral) and 20 degrees (cold) water temperatures, and also rested for 45 min (control). Energy expenditure (EE) was determined using indirect calorimetry before, during, and after each condition. Following exercise or rest, subjects had free access to a standard assortment of food items of known caloric value. EE was similar for the cold and neutral water conditions, averaging 505 +/ 22 (+/- standard deviation) and 517 +/- 42 kcal, respectively (P = NS). EI after the cold condition averaged 877 +/- 457 kcal, 44% and 41% higher (P < 0.05) than for the neutral and resting conditions, respectively. Cold-water temperature thus stimulated post-exercise EI. Water temperature warrants consideration in aquatic programs designed for weight loss. PMID- 15902989 TI - The influence of chokeberry juice supplementation on the reduction of oxidative stress resulting from an incremental rowing ergometer exercise. AB - The aim of the study was to investigate the influence of an increased intake of anthocyanins, contained in chokeberry juice, on the redox parameters in rowers performing a physical exercise during a 1-month training camp. The athletes were randomly assigned to receive 150 mL of chokeberry juice daily, containing 23 mg/100 mL of anthocyanins (supplemented group), or placebo (control group). Before and after the supplementation period, the subjects performed an incremental rowing exercise test. Blood samples were taken from the antecubital vein before each exercise test, 1 min after the test, and following a 24-h recovery period. After the supplementation period, TBARS concentrations in the samples collected 1 min after the exercise test and following a 24-h recovery period were significantly lower in the subjects receiving chokeberry juice than in the control group. In the supplemented group, glutathione peroxidase activity was lower in the samples collected 1 min after the exercise test, and superoxide dismutase activity was lower in the samples taken following a 24-h recovery, as compared to the subjects receiving placebo. These findings indicate that an increased intake of anthocyanins limits the exercise-induced oxidative damage to red blood cells, most probably by enhancing the endogenous antioxidant defense system. PMID- 15902990 TI - Influence of pre-exercise acidosis and alkalosis on the kinetics of acid-base recovery following intense exercise. AB - The purpose of this study was to measure the recovery kinetics of pH and lactate for the conditions of pre-exercise acidosis, alkalosis, and placebo states. Twelve trained male cyclists completed 3 exercise trials (110% workload at VO2max), ingesting either 0.3 g/kg of NH4Cl (ACD), 0.2 g/kg of Na+HCO3- and 0.2 g/kg of sodium citrate (ALK), or a placebo (calcium carbonate) (PLAC). Blood samples (heated dorsal hand vein) were drawn before, during, and after exercise. Exercise-induced acidosis was more severe in the ACD and PLAC trials (7.15 +/- 0.06, 7.21 +/- 0.07, 7.16 +/- 0.06, P < 0.05, for ACD, ALK, PLAC, respectively). Recovery kinetics for blood pH and lactate, as assessed by the monoexponential slope constant, were not different between trials (0.057 +/- 0.01, 0.050 +/- 0.01, 0.080 +/- 0.02, for ACD, ALK, PLAC, respectively). Complete recovery of blood pH from metabolic acidosis can take longer than 45 min. Such a recovery profile is nonlinear, with 50% recovery occurring in approximately 12 min. Complete recovery of blood lactate can take longer than 60 min, with 50% recovery occurring in approximately 30 min. Induced alkalosis decreases metabolic acidosis and improves pH recovery compared to acidodic and placebo conditions. Although blood pH and lactate are highly correlated during recovery from acidosis, they recover at significantly different rates. PMID- 15902991 TI - Assessment of the effects of eleutherococcus senticosus on endurance performance. AB - The use of nutritional ergogenic aids containing Eleutherococcus senticosus (ES), a plant which is also known as ciwujia or Siberian ginseng, is relatively common among endurance athletes. Eleutherococcus senticosus has been suggested to improve cardiorespiratory fitness (CF) and fat metabolism (FAM) and, therefore, endurance performance (EP). This article reviews the studies that evaluated the effects of ES during endurance exercise, three of which suggest that ES substantially improves CF, FAM, and EP. However, each of these reports contains severe methodological flaws, which seriously threaten their internal validity, thereby rendering hazardous the generalization of the results. On the other hand, 5 studies that used rigorous research protocols show no benefit of ES on CF, FAM, and EP. It is therefore concluded that ES supplementation (up to 1000 to 1200 mg/d for 1 to 6 wk) offers no advantage during exercise ranging in duration from 6 to 120 min. PMID- 15902992 TI - Comparison of erythrocyte and skeletal muscle creatine accumulation following creatine loading. AB - This study attempted to determine the relationship between creatine (Cr) accumulation in human skeletal muscle and erythrocytes following Cr supplementation. If a strong relationship exists, a blood test might provide a practical, less invasive alternative than muscle biopsy for evaluating cellular Cr accumulation. Eighteen active, but not well-trained males were supplemented with Cr (4 x 5 g/d) for 5 d. Muscle biopsies (vastus lateralis) were obtained pre and post-loading and analyzed for Cr, phosphocreatine (PCr), and total Cr (TCr) content. Venous blood was also drawn at these times to determine erythrocyte Cr concentrations. Muscle Cr, PCr, and TCr concentrations were elevated (P < 0.05) by 39.8%, 7.5%, and 20.1% respectively following supplementation. Erythrocyte Cr concentrations were also elevated (P < 0.01) following the loading period, although to a greater relative degree than tissue concentrations (129.6%). Pre- and post-loading erythrocyte Cr concentrations were poorly and nonsignificantly correlated with that observed in skeletal muscle. Further, loading-mediated increases in erythrocyte Cr concentrations were poorly correlated with elevations in muscle Cr (r = 0.07), PCr (r = 0.06) or TCr (r = 0.04) concentrations. Erythrocyte Cr concentrations can be augmented by 5 d of Cr supplementation, however, this elevation does not reflect that observed in skeletal muscle obtained by muscle biopsy. Consequently, erythrocyte response to Cr loading is not a reliable measure of skeletal muscle Cr/TCr accumulation. PMID- 15902993 TI - Role of recA/RAD51 family proteins in mammals. AB - DNA damage causes chromosomal instability leading to oncogenesis, apoptosis, and severe failure of cell functions. The DNA repair system includes base excision repair, nucleotide excision repair, mismatch repair, translesion replication, non homologous end-joining, and recombinational repair. Homologous recombination performs the recombinational repair. The RAD51 gene is an ortholog of Esherichia coli recA, and the gene product Rad51 protein plays a central role in the homologous recombination. In mammals, 7 recA-like genes have been identified: RAD51, RAD51L1/B, RAD51L2/C, RAD51L3/D, XRCC2, XRCC3, and DMC1. These genes, with the exception of meiosis-specific DMC1, are essential for development in mammals. Disruption of the RAD51 gene leads to cell death, whereas RAD51L1/B, RAD51L2/C, RAD51L3/D, XRCC2, and XRCC3 genes (RAD51 paralogs) are not essential for viability of cells, but these gene-deficient cells exhibit a similar defective phenotype. Yeast two-hybrid analysis, co-immunoprecipitation, mutation analysis, and domain mapping of Rad51 and Rad51 paralogs have revealed protein-protein interactions among these gene products. Recent investigations have shown that Rad51 paralogs play a role not only in an early step, but also in a late step of homologous recombination. In addition, identification of alternative transcripts of some RAD51 paralogs may reflect the complexity of the homologous recombination system. PMID- 15902994 TI - Selective COX-2 inhibition with different doses of rofecoxib does not impair endothelial function in patients with coronary artery disease. AB - In this study, we investigated the effects of both 25 and 50 mg daily doses of rofecoxib on the endothelial functions of patients with coronary artery disease (CAD). For this purpose, 34 patients with documented severe CAD and who were under aspirin treatment (300 mg/day) were randomized to receive 4 weeks of treatment with a placebo (n = 10, group I), rofecoxib 25 mg/day (n = 12, group II), and rofecoxib 50 mg/day (n = 12, group III). Brachial artery vasodilator responses were measured in order to evaluate endothelial function. The percentage of change in endothelial-dependent vasodilation in groups I, II, and III were similar at the baseline level and showed no significant change after treatment (6.2+/-3.9% vs. 5.9+/-3.1% and 5.8+/-3.3% vs. 5.6+/-3.8% and 6.1+/-4.5% vs. 5.8+/ 4.1%, respectively; P > 0.05). Compared with the baseline, endothelium independent vasodilatation, as assessed by nitroglycerine (NTG), remained unchanged after the treatment period (11.2+/-6.9% vs. 10.3+/-7.1% and 11.2+/-6.3% vs. 9.9+/-5.1% and 9.5+/-4.9% and 8.8+/-4.6%, respectively; P> 0.05). Treatment with both doses also showed no significant effects on high-sensitivity C-reactive protein (hs-CRP) levels and resting arterial diameters (P > 0.05). In conclusion, 4 weeks of treatment with standard and high doses of rofecoxib showed no significant effects on either endothelial-dependent or independent vasodilator response or plasma hs-CRP levels in patients with severe CAD taking concomitant aspirin. PMID- 15902995 TI - Determination of depression risk factors in children and adolescents by regression tree methodology. AB - We used a regression tree method (RTM) to determine risks of depression in children/adolescents. The survey records of 4,143 children/adolescents in a study based in Mersin, Turkey served as data in this study, and multi-step, stratified, and cluster sampling were used. Effects of 24 variables (sex, smoking, parental problems, etc.) were evaluated on depression scores. The Child Beck Depression Inventory (CBDI) was used to determine the level of depression. Subjects were into 12 different groups based on magnitudes of mean depression scores. The interactions among 7 variables determined to be risk factors are shown on a schema. The STATISTICA (ver.6.0) package program was used for all computations. Although traditional statistical methods have often been used for analysis in this field, such approaches are associated with certain disadvantages such as missing values, ignorance of interaction effects, or restriction of the shape of the distribution. To avoid such disadvantages, we therefore suggest the use of the RTM in studies involving numerical-based outcome variables and for the investigation of a large number of variables and it may be more effective than traditional statistical methods in epidemiological studies which determine risk factors. PMID- 15902996 TI - Clostridium perfringens foodborne outbreak due to braised chop suey supplied by chafing dish. AB - On February 13, 2002, a public health center in Hiroshima Prefecture, Japan, was notified that many individuals living at the Japan Maritime Self-Defence Force base had symptoms resembling those of food poisoning. Self-administered questionnaires requesting information regarding meal consumption and symptoms were distributed to all 281 members at the base. A case of the illness was defined as a member who had had watery or mucousy stool, or loose stool with abdominal cramps, more than twice a day after consuming dinner on February 12. Control of the illness was defined as a member with no symptoms. The dinner on February 12 was significantly associated with the illness (Mantel-Haenszel odds ratio: 3.59, 95% confidence interval: 1.06-12.20). A case-control study showed that, among the food supplied at dinner on February 12, the braised chop suey was significantly associated with the illness (odds ratio: 12.30, 95% confidence interval: 1.90-521.00). The braised chop suey had been stored in a chafing dish. An environmental investigation indicated that Clostridium perfringens (C. perfringens) in the chafing dish proliferated under an inappropriate heat retention temperature, and the contaminated braised chop suey could have caused the food poisoning. This study demonstrated that the recommended heat-retention temperature (over 65 degrees C) should be confirmed thoroughly. PMID- 15902997 TI - A c-KIT codon 816 mutation, D816H, in the testicular germ cell tumor: case report of a Japanese patient with bilateral testicular seminomas. AB - Mutations of the c-KIT gene have been reported not only in gastrointestinal stromal tumors and mast cell tumors, but also in testicular germ cell tumors (TGCTs). In the present study we employed polymerase chain reaction and DNA sequencing analysis to characterize the c-KIT gene in a 29-year-old Japanese patient with bilateral testicular seminomas. Direct sequence analyses revealed a single base substitution in exon 17 in one c-KIT allele, resulting in an amino acid substitution of D816H in this mutated allele. This mutation was found in the left, but not in the right, testicular seminoma. This is the first description of a c-KIT gene mutation in a Japanese patient with bilateral TGCT. The mutational analysis of the c-KIT gene seems to provide crucial information for managing TGCT patients not only in Europe but also in Japan. PMID- 15902998 TI - [Patients' rights and end of life]. PMID- 15902999 TI - [High-dose buprenorphine substitution during incarceration. Management of opiate addicts]. AB - OBJECTIVE: To describe the social and medical profiles of incarcerated (in detention or after sentencing) opiate addicts, whether or not they had already begun substitution treatment at arrival, and assess the impact of high-dose buprenorphine substitution therapy on the health of prisoners and the course of their incarceration. METHODS: A prospective survey was conducted on opiate addicts on admission to prison and after 2 months of incarceration, from December 2001 to February 2003, in 6 prison centres in the South East of France. RESULTS: During incarceration, no significant difference (other than in medical follow-up) appeared between the prisoners receiving substitution treatment and those who went through withdrawal on arrival. The first group differed from the second in several respects: their occupational history before incarceration was less stable, their history of drug addiction and incarceration was more serious (injection, psychotropic use, number of prior incarcerations, early age at first incarceration). The buprenorphine patients also differed in their more intense use of medical follow-up before incarceration. CONCLUSION: The impact of buprenorphine substitution therapy during incarceration could not be demonstrated, but prisoners receiving this treatment had a substantially different profile than those who were not receiving treatment when they arrived in prison. PMID- 15903000 TI - [Severe Streptococcus agalactiae infection of the diabetic foot. A deleterious role of Streptococcus agalactiae?]. AB - INTRODUCTION: Screening strategies among pregnant women have decreased the incidence of group B Streptococcus, which causes severe neonatal infections. The incidence of these infections has increased among diabetic patients, however. OBJECTIVES: To specify the characteristics of diabetic foot infections in which surgical samples have isolated one or several germs including group B Streptococcus, study its risk factors and determine its course. MATERIALS AND METHODS: We retrospectively evaluated the records of all patients admitted to the University Hospital of Geneva from January 1999 through October 2004, with diagnoses of severe foot infection (+/- osteomyelitis) documented during surgery. RESULTS: Twenty-five severe diabetic foot infections were identified, 21 with osteomyelitis. The most common risk factors were age older than 60 years (n=10), chronic renal failure (n=7), severe arteriopathy (n=6), and immune depression (n=2). Most lesions were classified as grade 3 or 4 of Wagner's classification. 80% of the surgical samples were polymicrobial. Blood cultures were positive in 4 patients, one in septic shock. Half the patients (n=13) underwent amputation, despite initially appropriate antibiotic treatment. No patients died but 3 relapsed. CONCLUSION: Group B streptococcal foot infections often occur in fragile patients with immune depression or severe arterial disease. Despite intensive antibiotic therapy and adequate debridement, amputation is often required in diabetic patients because of severe damage to the tissue and poor vascularization. PMID- 15903001 TI - [Terminal extubation in 5 end-of-life patients in intensive care units]. AB - INTRODUCTION: Decisions to withhold or withdraw active life support treatment in situations with no hope of improvement remain difficult for critical-care specialists and families; they are not always well understood by the public. This report describes terminal extubation, a particular method of withdrawing ventilator support. METHODS: This retrospective analysis examines the records of patients who died in our intensive care unit after a decision to withdraw active life support by stopping artificial ventilation. Extubation was proposed for patients with irreversible neurological damage and was always performed only after a standardized collective decision-making process. This process included three stages. In the initial phase, withdrawal of ventilator support was discussed at a department staff meeting. The meeting's conclusions were transcribed into the medical file, and the possibility of extubation was raised with the family during a planned interview. At least a 24-hour period of reflection was necessary before a new interview, and any opposition, hesitation or lack of understanding by the family at this first interview resulted in suspending the decision. The technical procedures for terminal extubation were also standardized. RESULTS: In 5 cases (4 men and one woman, with a mean age of 65 +/- 4 years), terminal extubation was decided in cooperation with the family, following an average of 3 interviews, 16 days after admission. All patients died within 3 days. DISCUSSION: So-called "terminal" extubation, very common in the United States, but much less so in France, reinforces the transparency of end-of life decisions in intensive care units and immediately makes tangible the end of the aggressive treatment for which critical-care specialists have been reproached. Since this first series of patients, extubation has been practiced in our department, principally in situations of irreversible neurological damage. PMID- 15903002 TI - [Systematic use in hospitals of a alcohol problems detecting questionnaire. The CAGE (cut down, annoyed, guilty, eye opener) questionnaire]. AB - OBJECTIVES: The CAGE questionnaire is considered a useful tool for detecting alcohol problems, especially in clinical populations. Its sensitivity exceeds 75% and its specificity 90%. Our objective was to assess performance of the French version of the CAGE (DETA) in identifying patients who meet the DSM-IV criteria and to discuss its usefulness in hospitals. METHODS: Data come from study of hospital morbidity in the Auvergne district of central France in 1998. Alcohol problems were diagnosed by physicians using the DSM-IV alcohol abuse or dependency criteria. For each diagnostic category, we calculated the performance of the CAGE questionnaire at a cut-off of 2 (CAGE 2+) and of 1 (CAGE 1+). RESULTS: The study included 5,452 patients--48.5% of them men--in acute- and intermediate-care facilities. CAGE 2+ had a sensitivity of 77% and a specificity of 94% among the diagnosed patients. It was more sensitive for patients diagnosed as alcohol-dependent than for alcohol abusers (61% vs 84%), but specificity was identical for both groups (94%). These results are similar to the properties of the English version. For a cut-off of 1 the CAGE had a sensitivity of 88% and a specificity of 87%. CONCLUSION: The French version of CAGE is a useful screening tool for alcohol abuse and dependance. Its systematic use in hospitals is desirable: a positive response to any item should attract the physician's attention and lead to further investigation. PMID- 15903003 TI - [Bronchiolitis obliterans organizing pneumonia following radiotherapy]. AB - INTRODUCTION: Pulmonary complications of radiotherapy are rare, but bronchiolitis obliterans organizing pneumonia (BOOP) is observed in 2.5% of cases. It can develop after radiation treatment of breast cancer as well as, more rarely, lung cancer, sarcoma, Hodgkin's disease or malignant thymoma. CASE: Ten months after radiotherapy for breast cancer, a 52 year-old woman developed migratory alveolar opacities outside the radiation field. Their improvement with corticosteroid treatment led to the diagnosis of BOOP. DISCUSSION: BOOP, which resembles infectious pneumonia, can develop 2-7 months after the end of radiotherapy and is seen especially in women aged 50-60 years with fever and coughs resistant to antibiotics. Dyspnea is far rarer. Imaging reveals patchy infiltrates with widespread bilateral, mobile lesions extended over and above the radiation field. Biopsy is required to confirm diagnosis; sections, which may or may not come from the radiation field, reveal the nonspecific granulomatous alveolar infiltrates typical of BOOP. Other causes should be eliminated (toxic, immune, iatrogenic or even idiopathic infection and recurrent early neoplastic relapse). Association with hormone therapy does not influence the course of BOOP. Outcome with corticosteroid treatment is excellent. PMID- 15903004 TI - [Carcinomatous meningitis as a rare presentation of gastric cancer]. AB - INTRODUCTION: Carcinomatous meningitis is a rare complication of gastric cancer. CASE: A 46 year-old man with an unremarkable medical history was hospitalized for treatment of suspected meningitis. Despite treatment, his clinical state rapidly worsened and he died without regaining consciousness, shortly after transfer to the ICU. Autopsy showed that he had anchorage-independent cell gastric carcinoma, with simultaneous lymphatic, pulmonary, cutaneous and meningeal metastases. DISCUSSION: The rising incidence of secondary meningeal lesions in the last two decades is probably associated with the treatment-related improvement in life expectancy. Prognosis is very grave, and median survival time is only several weeks. Carcinomatous meningitis is difficult to diagnosis, especially as the first sign of a primary tumor. This rare presentation of gastric cancer indicates diffuse metastatic spread and extremely poor prognosis. PMID- 15903005 TI - [Cushing's syndrome and adrenal adenoma. Two surprising associations]. AB - INTRODUCTION: Cushing's syndrome has a very low incidence (1-10 cases/million/year), and familial cases are even more rare. We report on two situations involving different causes of Cushing's syndrome. CASES: In the first case, we describe the case of a patient with an adrenal adenoma 20 years before the occurrence of Cushing's disease related to the pineal gland. In the second case, two members of the same family were diagnosed almost simultaneously with adrenal cortical adenoma (mother) and Cushing's disease (daughter). DISCUSSION: These cases lead us to consider the known causes of familial Cushing's syndrome, which were not found here. PMID- 15903006 TI - [Drug hypersensitivity syndrome with lamotrigine two cases in elderly]. AB - INTRODUCTION: Lamotrigine is an antiepileptic agent used in a wide range of seizure disorders among the elderly. In rare cases, it can induce a drug hypersensitivity syndrome (DHS). CASES: DHS due to lamotrigine was observed in two patients (85 and 91 years-old). The first case was characterized by febrile erythema with eosinophilia and the second by isolated fever. Lamotrigine was replaced by sodium valproate for one patient. Both patients had satisfactory outcomes after discontinuing lamotrigine. DISCUSSION: Lamotrigine can induce DHS that is clinically and biologically similar to the DHS observed with standard antiepileptic drugs. It may involve concomitant or consecutive treatment with other antiepileptic agents, in particular, valproate acid, which decreases lamotrigine clearance. Potentially serious and even fatal, these adverse reactions are to be feared in a population with multiple comorbidities and can cause harmful diagnostic mistakes. They are especially fearsome in geriatric populations with high epilepsy rates. PMID- 15903007 TI - [Malaria in splenectomized patients. Three fatal cases]. AB - INTRODUCTION: Splenectomy, the surgical removal of the spleen, is increasingly practiced in malaria-endemic tropical countries. The procedure leaves patients more susceptible to serious bacterial and parasitic infections, including malaria. CASES: We report here three fatal cases of malaria from P. falciparum in splenectomized patients. One operation followed abdominal trauma, while the other two were performed to treat one large-cell and one B-cell lymphoma. Despite prolonged intravenous quinine treatment all three patients died. DISCUSSION: Malaria in splenectomized patients can be very serious and is fatal in half the cases transmitted by P. falciparum. Permanent chemoprophylaxis is required for these patients when they travel to or live in endemic areas. PMID- 15903008 TI - [Nocardia farcinica brain abscess associated with a pulmonary embolism in an immunocompetent patient]. AB - INTRODUCTION: Nocardiosis is a rare, generally systemic infection that occurs in immunocompromised patients. We report a case of Nocardia farcinica primary brain abscess in an immunocompetent patient, unusually associated with a pulmonary embolism. OBSERVATION: A 62 year-old woman without medical past was hospitalised because of a Bravais-Jacksonian type of seizure. The clinical and radiological profile was unspecific. The diagnosis was made on identification of the microorganism. The patient improved after craniotomy with excision of the abscess and appropriate antibiotic therapy for four months. DISCUSSION: The N. farcinica species is distinct from others by its high degree of antibiotic resistance its virulence. We insist on the need to identify not only the microorganism but also the species concerned because this may influence the treatment strategy. PMID- 15903009 TI - [The project of a law concerning patients' rights at the end of life]. AB - A bill about Patients' rights and the end of life was adopted unanimously by the National Assembly on November 30, 2004. Article 1 provides that the physician is not obligated to continue treatment "when the latter appears futile, disproportional and has no effect other than artificially maintaining the patient's life". Article 2 recognizes that painkillers administered at high doses to terminally ill patients may as a side effect "shorten the patient's life". Conscious patients and the families of unconscious patients must be so informed, and the discussion must be mentioned in the patient's file. Physicians must respect their patients' refusal of treatment, even in life-threatening situations. On the other hand, patients who are not terminally ill must await the expiration of a mandatory waiting period before the physicians must comply with their wishes to refuse care. Article 3 specifies that any treatment can be withheld or withdrawn, including artificial nutrition. The law specifies that it is applicable in 4 different sets of situations: for patients who are or are not terminally and for those who are or are not conscious. Article 9 makes clear that futile treatment of unconscious patients can be withheld or withdrawn. Article 7 specifies that adults may draw up advance directives to indicate their wishes for their end of life and their desires regarding the withholding or withdrawal of treatment. Withholding or withdrawing active treatment is authorized under 3 conditions: the consent from the patient, or if he or she is unconscious, the approval of the health-care proxy, or a family member or close friend if the patient is unconscious; the inclusion of the decision in the patient's medical file, and a group decision-making process. PMID- 15903010 TI - [Sexual dysfunction in schizophrenics patients, the role of antipsychotics]. AB - Although rarely reported spontaneously, sexual dysfunction is frequent in patients with schizophrenia. Sexual dysfunctions may be related to schizophrenia, to neuroleptic use and to psychiatric comorbidities, especially depression and addiction. Sexual dysfunctions induced by antipsychotics are related to their effect on the adrenergic and dopaminergic histamine receptors, in particular their blockage of the dopaminergic receptors of the tuberoinfundibular tracts, which causes excess prolactin secretion. PMID- 15903011 TI - [Pathophysiology of infection on orthopedic biomaterials]. AB - Orthopedic biomaterials are foreign bodies and the molecular architecture of their surfaces provides a point of attachment for bacteria. This adherence is made possible through the interaction of the protein interface and the bacterial adhesins. Bacterial colonies use slime and biofilm as means of protection. The development of bacteria towards a reversible state of stationary growth or microcolony variants permits their survival. Microparticles released by biomaterials cause the chronic inflammation associated with the aseptic loosening of prostheses. Some bacterial sub-populations develop transitory resistance to bactericidal antibiotics in the presence of these materials. PMID- 15903012 TI - [Pathophysiology of acne]. AB - Acne is a disease that develops in pilosebaceous follicles. Acne was long considered was an infectious disease related to Propionibac-terium acnes, but studies show that the production of inflammatory substances in situ was much more important to the action of this bacterium than was simple infection. It was once thought that only the androgens could stimulate the seba-ceous gland, but today we know that neuromediators can also stimulate sebum production. This finding provides physiological support to the clinical observation that stress can induce acne. PMID- 15903013 TI - [The treatment of acne]. AB - The treatment of acne is based on local and systemic treatments. It must be adapted to the type of acne and take some prognostic factors into account. Because of the possibility of drug resistance, systemic and local antibiotics should be used for only limited periods and preferably as part of combined treatments. Prescription of isotretinoin to women must follow strict rules. Some new-generation contraceptive pills are an interesting alternative for acne treatment in women. Zinc, which basically targets inflammatory acne, is a possible option for pregnant women and during the summer. Cosmetic care is also a fundamental part of acne treatment. PMID- 15903014 TI - [Obturator hernia, a rare cause of mechanical occlusion of the small intestine]. PMID- 15903015 TI - [The re-emergence of syphilis]. PMID- 15903016 TI - [Vaccinations, from theory to practice]. PMID- 15903017 TI - [Cardiovascular risks and cyclooxygenase-2 inhibitors]. PMID- 15903018 TI - Scandinavian Journal of Rheumatology 1955-2005: development and state of the Journal on its 50th anniversary. AB - The Scandinavian Society for Rheumatology was founded in Copenhagen in September 1946 and is still actively stimulating cooperation among rheumatologists in the Scandinavian countries by organising biannual Scandinavian congresses, and by circulating an international scientific publication, the Scandinavian Journal of Rheumatology, now celebrating its 50th anniversary. We hope that the newly implementation of web-based submission of manuscripts and peer reviewing, together with the new cover and change in the Journal's contents, will be of benefit to our authors, reviewers, and readers. At the Editorial Office we express our hope for a continuous and fruitful collaboration with the many people involved in the Scandinavian Journal of Rheumatology. Our thanks are due to those who made it possible over the past 50 years to publish a journal of high scientific quality within the field of rheumatology. PMID- 15903019 TI - Aspects of science in Scandinavian rheumatology--the making of a specialty. AB - The five Scandinavian countries Denmark, Finland, Iceland, Norway, and Sweden have all contributed essential and unique knowledge to the science of rheumatology. The origins have varied between the countries. Bridges from basic science and international contacts have been essential for the quality and vitality of a small specialty in small countries. Inter-Scandinavian links as manifested in congresses and in the Journal are the visible superstructure of a far deeper and fertile common culture. PMID- 15903020 TI - The molecular basis for the effectiveness, toxicity, and resistance to glucocorticoids: focus on the treatment of rheumatoid arthritis. AB - Glucocorticoids (GCs) have powerful and potent anti-inflammatory and immunomodulatory effects in rheumatoid arthritis (RA) and many other diseases. These effects are mediated by up to four different mechanisms of action: cytosolic glucocorticoid receptor (cGCR)-mediated classical genomic and rapid non genomic effects, membrane-bound glucocorticoid receptor (mGCR)-mediated non genomic effects and non-specific non-genomic effects. On the basis of this detailed knowledge of mechanisms there are currently interesting approaches being considered that may lead to the development of GC drugs and GCR ligands with an improved benefit to side-effect ratio. Another interesting field of GC research is the phenomenon of GCR resistance. Several different mechanisms may mediate this phenomenon; among them are alterations in number, binding affinity, or phosphorylation status of the GCR. Other mechanisms of GC resistance being investigated are polymorphic changes and/or overexpression of (co-)chaperones, the increased expression of inflammatory transcription factors, overexpression of the GCR beta isoform, the multidrug resistance pump, and an altered mGCR expression. PMID- 15903021 TI - TNF-alpha -308 promoter polymorphism in patients with rheumatoid arthritis. AB - OBJECTIVES: Rheumatoid arthritis (RA) is a chronic inflammatory disease in which tumour necrosis factor-alpha (TNF-alpha) plays an important role. There are, however, controversial reports that TNF-alpha promoter polymorphism may be an independent marker of susceptibility and severity of RA. The aim of the present study was to examine the TNF-alpha -308 promoter polymorphism in patients with RA. METHODS: We examined 91 patients with RA diagnosed according to the criteria of the American College of Rheumatology. Polymerase chain reaction (PCR) amplification was used for analysis of the polymorphism at position -308 in promoter of TNF-alpha gene. RESULTS: Distribution of TNF-alpha genotypes in RA patients did not differ from that in control subjects. Moreover, there was no association between TNF-alpha genotypes and age at disease diagnosis, disease activity in global physician's assessment, and joint and extra-articular involvement. There was also no correlation between TNF-alpha polymorphism and disease activity measures, including erythrocyte sedimentation rate (ESR), CRP, number of swollen and tender joints, and morning stiffness duration. CONCLUSIONS: We suggest that TNF-alpha -308 promoter polymorphism is not a genetic risk factor for RA susceptibility and severity. PMID- 15903022 TI - Comparison of dual X-ray absorptiometry (DXA), digital X-ray radiogrammetry (DXR), and conventional radiographs in the evaluation of osteoporosis and bone erosions in patients with rheumatoid arthritis. AB - OBJECTIVE: To compare the relationship between bone mineral density (BMD) in the metacarpal bones and forearm measured by dual X-ray absorptiometry (DXA) and digital X-ray radiogrammetry (DXR) and radiological alterations in patients with early and established rheumatoid arthritis (RA). PATIENTS AND METHODS: In each of the three disease duration groups, 11 female RA patients were included. The patients were further divided into two groups according to bone erosions. BMD in the metacarpals was evaluated by DXA and DXR. RESULTS: A significant relationship between DXA-BMD and DXR-BMD was observed. DXR-BMD and the individually combined cortical thickness (CT) of the metacarpo-phalangeal (MCP) joints were related to disease duration and erosions. Patients with erosive disease had lower values of age- and sex-adjusted BMD measured with DXA, but most significantly with DXR. CONCLUSION: DXR appears to be a more sensitive method than DXA in detecting early bone loss in patients with RA. The relationship of DXR-BMD to disease duration and bone damage indicates that the DXR method may be useful in the evaluation of disease activity and progression. PMID- 15903023 TI - Urinary excretion of alpha-GST and albumin in rheumatoid arthritis patients treated with methotrexate or other DMARDs alone or in combination with NSAIDs. AB - OBJECTIVE: To evaluate the effect of methotrexate (MTX) and other disease modifying anti-rheumatic drugs (DMARDs) alone or in combination with non steroidal anti-inflammatory drugs (NSAIDs) on the urinary excretion of alpha glutathione S-transferase (alpha-GST) and albumin in rheumatoid arthritis (RA) patients. METHODS: Nineteen RA patients starting treatment with MTX were followed for 1 year with measurements of urinary alpha-GST, urinary albumin, and urinary and plasma creatinine at the start of treatment, and after 16, 28, and 52 weeks. A larger group of RA patients (n = 72) undergoing long-term treatment with different DMARDs was compared with 79 healthy controls regarding urinary alpha GST and albumin. alpha-GST was quantified by an enzyme immunoassay. Urine albumin was measured turbidimetrically. RESULTS: The urine-alpha-GST/urine-creatinine ratio and the urine-albumin/urine-creatinine ratio did not change during 52 weeks of treatment with MTX. The long-term DMARD-treated RA patients and the healthy controls were comparable with regard to the urine-alpha-GST/urine-creatinine ratio and the urine-albumin/urine-creatinine ratio. All patients had preserved renal function as assessed by plasma creatinine, and none had proteinuria using urine dipstick methods. CONCLUSION: DMARD-treated RA patients with normal serum creatinine had no detectable renal injuries assessed by the urinary excretions of alpha-GST and albumin. PMID- 15903024 TI - Routine database registration of biological therapy increases the reporting of adverse events twentyfold in clinical practice. First results from the Danish Database (DANBIO). AB - OBJECTIVE: To present from the Danish Database for Biological Therapies in Rheumatology (DANBIO) the frequencies and types of adverse events as well as risk factors during treatment with biological agents in clinical practice. METHODS: Adverse events during the first 2 years of clinical use of biological agents in Denmark were reported to the nationwide DANBIO and compared to the mandatory reports to the Danish Medicines Agency. RESULTS: Almost 90% of the patients treated with biological agents were registered in the DANBIO, and the database picked up 20 times as many adverse events as the Danish Medicines Agency. Infections and hypersensitivity reactions were the most prevalent adverse events. Age, disease duration, and previous number of disease-modifying anti-rheumatic drugs (DMARDS) were found to be risk factors for bacterial infections. CONCLUSION: A routine-based Danish database for biological therapies covers approximately 90% of patients and improves the reporting of adverse events. PMID- 15903025 TI - Reproduction and gynaecological manifestations in women with primary Sjogren's syndrome: a case-control study. AB - BACKGROUND: The impact of primary Sjogren's syndrome (pSS) on reproduction and gynaecological manifestations has seldom been explored. AIM OF STUDY: Assess gynaecological aspects, gynaecological interventions, and use of contraceptives in a population of pSS-patients versus controls. METHODS: In a case-control study, 58 pSS-patients and 157 controls answered a self-administered questionnaire, covering demographic data, reproductive events, gynaecological problems, and gynaecological interventions. RESULTS: Significantly more patients than controls reported episodes of amenorrhoea lasting for more than 3 months, and more patients suffered from menorrhagia/metrorrhagia compared with controls (54.5% versus 35.7%; p = 0.012). Complaints of vaginal dryness were common among the patients (52.9% versus 28.3%; p = 0.005). Endometriosis was reported to occur more frequently in the patients (8.5% versus 2.1%; p = 0.03), and 6.3% of pSS patients reported having undergone surgical intervention for endometriosis versus 0.7% of the controls (p = 0.009). Positive information about surgery for endometriosis correlated with the presence of the autoantibodies anti-SSA (r = 0.322; p = 0.008) and anti-SSB (r = 0.313; p = 0.01). Among the pSS-patients, 5.9% had chosen not to have children due to the disease, but there was no indication of reduced fertility as judged by the number of pregnancies. CONCLUSION: Patients with pSS reported more gynaecological problems than controls, including vaginal sicca symptoms, endometriosis, several episodes of amenorrhoea, and menorrhagia/metrorrhagia. PMID- 15903026 TI - The serological pattern of autoantibodies to the Ro52, Ro60, and La48 autoantigens in primary Sjogren's syndrome patients and healthy controls. AB - OBJECTIVE: Sjogren's syndrome (SS) is characterized by exocrine secretion dysfunction. Hallmarks of the chronic autoimmune disease are cellular infiltration of the exocrine glands and the presence of serum autoantibodies against Ro and La. The purpose of this study was to perform a detailed characterisation of the serological pattern against the Ro and La autoantigens in terms of antigen specificity and antibody isotype. METHODS: Serum samples from 100 patients with primary SS and 100 matched healthy controls were tested by enzyme-linked immunosorbent assay (ELISA) with recombinant human Ro and La proteins as antigens. RESULTS: There were higher frequencies of Ro and La positive serum in the SS patients than in the control sera, and the titres were higher in the positive sera from SS patients than the controls. The SS patients often had antibodies against two or three of the antigens tested, while the positive control sera often reacted against only one of the autoantigens. The SS patients had a broader immunoglobulin isotype repertoire in their autoantibodies while the controls when positive usually had one antigen specific isotype. CONCLUSION: We found a distinct and significant difference in the serum antibody specificity and immunoglobulin isotype pattern between SS patients and matched controls. This variance may point to different mechanisms by which these autoantibodies are generated. PMID- 15903027 TI - MEFV gene is a probable susceptibility gene for Behcet's disease. AB - OBJECTIVE: Behcet's disease (BD) is a rare, chronic, multisystem inflammatory disorder. The prevalence of BD is higher in the Middle Eastern and Mediterranean populations. Another chronic inflammatory disease, familial Mediterranean fever (FMF), is also known to be highly prevalent in these populations. The prevalence of BD is higher in the FMF patient population than in populations known to be rich in BD . Both BD and FMF have some pathophysiological features in common and they result from inappropriate activation of neutrophils. Clinical manifestations of both diseases can mimic each other and the coexistence of both diseases in the same patient has been reported. Given that BD and FMF have similar pathophysiological, epidemiological, and clinical features, we hypothesized that the gene responsible for FMF, MEFV, may also play a role in the pathogenesis of BD. METHODS: Forty-two BD patients who had no symptoms and family history for FMF and 66 healthy controls were screened for common MEFV gene mutations (E148Q, M680I, M694V, and V726A). RESULTS: Fifteen patients (36%) displayed MEFV mutations (nine M694V, five E148Q, and one M680I) and mutation rates were significantly elevated compared to 66 (11%) healthy controls (p = 0.0034). CONCLUSION: The occurrence of frequent MEFV mutations in BD patients suggests that the MEFV gene is involved in the pathogenesis of Behcet's disease. PMID- 15903028 TI - Knee osteoarthritis and body mass index: a population-based case-control study. AB - OBJECTIVE: It is well established that overweight is related to osteoarthritis of the knees. The aim of this study was to investigate the risk of knee osteoarthritis for men and women in relation to body mass index (BMI) within the normal weight range and to assess the effect of former versus current weight. METHODS: A population-based case-control study was carried out in the southern part of Sweden, including 825 cases with X-ray verified femorotibial osteoarthritis and 825 age-, sex-, and county-matched population controls. Mailed questionnaire data on weight, height, and confounding factors (heredity, smoking, knee injuries, and physical activity) were collected and analysed using logistic regression models. The response frequency was 89%. RESULTS: Mean age of the participants was 63 years, and 57% were women. The adjusted risk of knee osteoarthritis was increased fourfold in men with a current BMI 23 to < 25 kg/m2 as compared to men with BMI < 23 kg/m2 (OR 4.0, 95% CI 1.7-9.5). The commensurate risk for women was 1.6 (95% CI 0.9-3.1). BMI at 30 years of age was similarly related to knee osteoarthritis. CONCLUSION: A moderate increase in BMI, within the normal weight range, was significantly related to knee osteoarthritis among men. Overweight at any time was related to knee osteoarthritis. PMID- 15903029 TI - Macrophagic myofasciitis unrelated to vaccination. AB - Macrophagic myofasciitis (MMF) is known to be closely associated to vaccination containing aluminium as an adjuvant. Here we describe a case of MMF unrelated to vaccination, and presented as focal tender muscle in a 59-year-old woman. Magnetic resonance imaging showed evidence of unilateral myofasciitis involving muscles and fascias of the vastus lateralis. Open biopsy showed a unique pathological pattern of MMF. Our case suggests that other causes unrelated to vaccination need to be investigated in the pathogenesis of MMF, and MMF could be considered in the differential diagnosis of patients with focal muscle tenderness. PMID- 15903030 TI - Strong expression of Bcl-2 in pigmented villonodular synovitis of the knee with aggressive clinical behaviour. AB - OBJECTIVE: To determine the expression of bcl-2, p53, and caspase 3, and measure the Ki-67 proliferation index as well as DNA content and DNA fragmentation in a case of diffuse pigmented villonodular synovitis (PVNS) of the knee with aggressive clinical behaviour. METHODS: Expression of p53, Bcl-2 and Ki-67 was investigated using immunohistochemistry. In addition, multiparametric flow cytometry was performed for expression of p53, bcl-2, and caspase 3, as well as analysis of DNA content and distribution of cell cycle phases. DNA fragmentation was detected by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL). RESULTS: A strong cytoplasmic positivity for Bcl-2 protein, a key factor in regulation of apoptosis, was found in the majority of proliferating synovial cells. No apoptotic cell fraction was found by analysis of DNA content. DNA fragmentation was observed in 6.8% of cells. No elevated expression of p53 and caspase 3 was detected. CONCLUSION: Our results indicate a possible role of dysregulation of apoptosis in this case of PVNS. This aspect in the pathogenesis of PVNS should be clarified in further studies. PMID- 15903031 TI - Peripheral gangrene in adult-onset Kawasaki disease. AB - Kawasaki disease (KD) is an acute vasculitis characterized by marked tropism of the coronary vessels. Usually a childhood disease, KD can occasionally be observed in adults. We report a case of adult-onset KD that presented as a prolonged fever of unknown origin with subsequent development of severe vasculitis, manifested by coronary aneurysms and peripheral gangrene of the lower limbs. Therapy with intravenous immunoglobulins, corticosteroids, aspirin, anticoagulants, and prostacyclin analogue resulted in rapid improvement in the patient's condition but he required partial distal amputation for irreversible gangrene. Peripheral ischaemia leading to gangrene is a very rare feature of KD vasculitis with only 19 cases previously published in the Medline-indexed literature. The outcomes of those patients were poor, being either fatal or requiring distal amputation. All prior cases affected young children less than 1 year old and, to the best of our knowledge, the present case is the first description of peripheral gangrene in adult-onset KD. PMID- 15903032 TI - Are infections increased in rheumatoid arthritis (RA) prior to diagnosis? Results of a case control study of RA compared to non-inflammatory musculoskeletal disorders. PMID- 15903033 TI - Effects of interferon-alpha treatment on serum IL-6, IL-8, TNF-alpha and soluble TNF-alpha receptors in Behcet's disease. PMID- 15903034 TI - Cerebral demyelination in association with TNF-inhibition therapy in a 5-year-old girl with aseptic meningitis as the first symptom of Still's disease. PMID- 15903035 TI - [Pulmonary infection caused by non-tuberculous mycobacteria in two patients with bronchiectasis]. AB - Two patients, a woman aged 67 years and a man aged 80 years, had chronic cough among other respiratory symptoms. In the woman, chest radiograph and CT-scan revealed partial atelectasis of the middle lobe and bronchiectasis. In the man, an interstitial pattern was seen on chest radiograph, and CT scan showed diffuse bronchiectasis. In both the man and the woman, non-tuberculous mycobacteria were identified (Mycobacterium avium complex and Mycobacterium abscessus, respectively). Treatment was successful in both patients. Non-tuberculous mycobacteria can cause considerable pulmonary infection in patients with bronchiectasis. PMID- 15903036 TI - [The unreasonableness of prostate-cancer screening and the ethical problems pertaining to its investigation]. AB - Since the early 1990s, screening with prostate-specific antigen (PSA) testing has increased the incidence of prostate cancer. Any decrease in mortality will not be seen for at least a decade, due to the long natural history of prostate cancer. Death due to prostate cancer is rare, while the prevalence oflocalised tumours is high. The prognosis of these early-detected localised tumours is uncertain, because most patients will die from other causes. Complications of prostate cancer therapy are common, with high rates of impotence, incontinence and gastrointestinal problems after prostatectomy or radiotherapy. Randomised trials of prostate-cancer screening, notably the 'European randomised screening for prostate cancer' (ERSPC) trial, began with the consent of ethical committees. There is a real uncertainty regarding the benefits of prostate-cancer screening. However, it is clear that these benefits are limited, because prostate-cancer death is rare before the age of 75 years. There is no real uncertainty about the harms of prostate-cancer screening. High prevalence and high rates of treatment complications deduct many disease- and disability-free years from the eligible population (men aged 55-74 years). Therefore, there has been no real uncertainty over the balance of harms and benefits in prostate-cancer screening trials. Days may be added to old age, at the cost of months of disease- and disability-free living. It is not in the best interest of eligible men to participate in these trials. Randomised trials evaluating prostate-cancer screening violate in principle and practice the Helsinki Declaration of the rights of human subjects in medical research. PMID- 15903037 TI - [Diagnosis based on prostate needle biopsy: inadequate correlation between pathologic results and clinical course for individual prognosis]. AB - Since the introduction of serum testing for prostate-specific antigen (PSA) in 1990 for the early detection of prostate cancer, the number of men undergoing a prostate needle-biopsy procedure has increased dramatically. In order to highlight the significance of the various diagnostic outcomes of a prostate needle biopsy, the pathological findings from needle biopsies were compared with those from samples taken during radical prostatectomy and in follow-up biopsies, using data from the 'European randomised screening for prostate cancer' (ERSPC) trial. In men with an elevated PSA value and a benign or negative needle-biopsy result, 10-15% were found to have prostate cancer in follow-up biopsies. In men with a needle-biopsy diagnosis of adenocarcinoma, 29% were found to have questionable histopathological characteristics or minimal cancer that did not require therapy. The incidence of minimal cancer increased to 70% among men with a needle-biopsy diagnosis of focal carcinoma, i.e. a small focus (< 3 mm diameter) of well-differentiated adenocarcinoma, based on one biopsy from a series of six. In men with a needle-biopsy diagnosis of 'suspected malignancy' 36.5% were found to have prostate cancer in follow-up biopsies. In men with a needle-biopsy diagnosis of 'high grade prostatic intra-epithelial neoplasia' 13% were found to have prostate cancer in follow-up biopsies. This percentage was not significantly higher than the percentage of cancers detected after an initially benign biopsy outcome. To avoid over-treatment of a substantial number of men who lack symptoms of prostate cancer but are diagnosed on the basis of biopsy results, it is vital that clinical/pathologic parameters are developed and validated that can help in deciding whether to initiate curative treatment immediately. PMID- 15903038 TI - [The investigation of gene-environment interaction using case-control comparisons]. AB - Many chronic diseases are caused by the interaction of genes and environmental factors. Genetic epidemiologic research seeks to elucidate the role of genetic factors and their interaction with environmental factors in the occurrence of disease. Gene-environment interaction can be assessed using different types of case-control comparisons. The classic case-control design is suitable to investigate the association between multiple genes and environmental factors. Results can be presented in a two-by-four table. In a case-parents design, the genotype of each case is compared with the genotype of a fictitious control formed by the non-transmitted alleles from each parent. In a case-only design, the relationship between a genotype and an environmental factor is examined in a population of affected cases only. Both the case-parents and the case-only designs assume independence between genotype and environmental factors in the control group. PMID- 15903039 TI - [Diagnostic image (237). A man with a swelling on his left buttock. Bednar tumor]. AB - A 42-year-old man was referred because of a pigmented dermatofibrosarcoma protuberans, a so-called Bednar tumour, on his left buttock. It was radically excised. PMID- 15903040 TI - [Large variation in indications for internal fixation or arthroplasty in displaced femoral neck fracture; results of a questionnaire distributed among general surgeons in 20 hospitals]. AB - OBJECTIVE: To describe the treatment protocols for displaced femoral neck fractures in all 8 university hospitals (UH) and 12 general hospitals (GH). DESIGN: Descriptive; questionnaire. METHOD: Questionnaires were distributed to general surgeons who also perform traumatology surgery. They were requested to give succinct answers to questions about local protocol for the maximum permissible time interval between hip trauma and operation, indications for internal fixation and arthroplasty, operative technique and postoperative degree of weight-bearing in patients over 60 years of age with a displaced femoral neck fracture. RESULTS: Internal fixation and arthroplasty were performed within 24 and 48 hours respectively in 95% of all hospitals. A biological upper age limit of between 65 and 80 years old was the most commonly quoted indication for internal fixation in 70% of all hospitals. In 83% of GH dementia was considered an indication for arthroplasty as opposed to 0% in UH. Poor bone quality, immobility, comminution and inadequate reduction were incidentally quoted indications for arthroplasty. Rheumatoid arthritis, arthrosis and pathological fracture were contra-indications for internal fixation in all hospitals. Operative techniques for internal fixation and arthroplasty were similar in both UH and GH. After internal fixation, full weight-bearing was recommended in all UH and partial weight-bearing in 7 (58%) of GH. Following arthroplasty all protocols prescribed full weight-bearing. CONCLUSION: The variation in indications for internal fixation or arthroplasty reflects the lack of studies that demonstrate clearly which patient can be treated optimally with which treatment modality. There were few differences in the operative techniques of internal fixation and arthroplasty between the hospitals. PMID- 15903041 TI - [No arguments to support an age limit for men entering an in vitro fertilisation or intracytoplasmic sperm injection programme]. AB - OBJECTIVE: To identify medical and psychosocial risks that could arise from allowing older men to father children through in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI). DESIGN: Literature review. METHOD: The databases Medline, Current Contents weekly, Current Contents archives, and PsycINFO were searched over the period 1970--June 2004 for articles with data on age limits for men entering IVF/ICSI programmes. The following inclusion criteria were used: availability in the Netherlands and written in English or Dutch. RESULTS: Although sperm quality decreases with age, men remain fertile up to an advanced age. The risks of having children with autosomal dominant disorders or chromosomal defects increase slightly, but the individual chance is extremely small. Studies on the psychological development of children with fathers aged > 50 years are lacking. Extrapolation from other studies indicates that growing up with an older father has no negative influence on child development. Although older fathers have a greater chance of dying sooner, the absence of the father does not contribute significantly to psychological problems in offspring later in life. CONCLUSION: There are no medical or psychosocial arguments to support an age limit for men entering an IVF/ICSI programme. PMID- 15903042 TI - [Posture-dependent headache due to the spontaneous hypotension syndrome]. AB - A 40-year-old woman and a 47-year-old man presented with acute posture-dependent headache. A spontaneous intracranial hypotension syndrome was diagnosed in both cases. MRI with a gadolinium contrast medium revealed staining of the pachymeninges. In the woman, cisternography revealed leakage of spinal fluid at the level of the cauda equina. Neither an infusion of caffeine nor an epidural blood patch helped, but the symptoms disappeared spontaneously. In the man, cisternography indicated leakage at the level of the 3rd thoracic vertebra. The symptoms disappeared rapidly after treatment with a local blood patch. Posture dependent headache is typical for the intracranial hypotension syndrome. The headache is usually relieved by lying down and aggravated by standing up, but the reverse has also been reported. This headache can develop in a short time, sometimes acutely, and may persist continuously. The syndrome is usually caused by leakage of cerebrospinal fluid due to rupture of the dura mater, which may occur spontaneously. The diagnosis can be established by gadolinium MRI, revealing a striking pattern of diffuse pachymeningeal enhancement. Subdural fluid accumulations may also be seen. Indium-pentetreotide cisternography can often localise the spinal fluid leak. Intravenous caffeine and the application of an autologous epidural blood patch are possible treatment options, but spontaneous recovery may also occur. PMID- 15903043 TI - [Headache and impaired consciousness due to the spontaneous intracranial hypotension syndrome]. AB - A 51-year-old man presented with a 6-week history of progressive headache, confusion and ataxic gate. The symptoms were not preceded by trauma or lumbar puncture. A CT-scan of the brain revealed bilateral subdural fluid accumulation and hyperdensities in the subarachnoid space. In view of the signs of a subarachnoid haemorrhage, angiography was performed but showed no indications of an aneurysm. An MRI-scan of the head revealed abnormalities in line with intracranial hypotension. CT-myelography of the whole spine revealed a cerebrospinal fluid leak at the level of the fifth and sixth thoracic vertebrae. The patient recovered completely after placement of an epidural blood patch at this level. Spontaneous intracranial hypotension shows clinical similarities with the symptoms following a lumbar puncture. In most cases it can be treated by conservative measures. However, invasive measures are sometimes necessary to close the defect in the meninges. PMID- 15903044 TI - [The pharmaceutical industry under discussion]. AB - The criticism of the pharmaceutical industry is growing while the confidence of the general population seems to be decreasing. The industry functions on the boundary between two worlds: the world of healthcare as a social system and that of a worldwide competitive industry providing employment to nearly 600,000 people in Europe. The two worlds are connected by clinicians who are engaged in the development of new drugs. The conflict between making a profit and providing care also exists in healthcare and science. The criticism should be taken seriously. The solution lies in the ability of the industry to purify itself and not in a new role for government or new regulations. PMID- 15903045 TI - [Lithium, a potentially dangerous drug]. PMID- 15903046 TI - [Lithium, a potentially dangerous drug]. PMID- 15903047 TI - [Lithium, a potentially dangerous drug]. PMID- 15903048 TI - [Lithium, a potentially dangerous drug]. PMID- 15903049 TI - [Lithium, a potentially dangerous drug]. PMID- 15903050 TI - [Knaves in white -- when the physicians do evil]. PMID- 15903051 TI - [How unhealthy is our environment? Fine dust costs six months, smoking six years]. PMID- 15903052 TI - [Rehydration of children with vomiting diarrhea: better infusion than nasal tube]. PMID- 15903053 TI - [Point value stability and trend increase: all only a Fata Morgana for the dying of thirst]. PMID- 15903054 TI - [Typical nitro-positive angina pectoris attacks -- nevertheless the coronary angiography was inconspicuous]. PMID- 15903055 TI - [Treat insulin resistance and look out for mental stress. New strategies in the prevention of type 2 diabetes]. PMID- 15903056 TI - [Preventing type 2 diabetes: what does dietary fiber achieve?]. AB - On the basis of the currently available data, an increase in the uptake of indigestible roughage can be recommended as a means of reducing the risk of developing type 2 diabetes and cardiovascular diseases. In concert with other factors of a healthy lifestyle--including physical exercise, abstention from nicotine, the consumption of only moderate amounts of alcohol, reducing the intake of saturated fats, and--the main factor--ensuring that overweight does not develop, this dietary modification would appear to be an effective measure for avoiding most cases of type 2 diabetes. PMID- 15903058 TI - [Depression--an important obstruction to the treatment of diabetes]. AB - In diabetics, depressive states represent a major impediment to treatment. This applies equally to persons with depressive mood (subclinical) and patients with clinically manifest depression. Particular consideration should be given to the possibility of depression in patients with emotional problems associated with the disease, recurrent episodes of severe hypoglycemia, and sequelae. The diagnosis is relatively easy to establish with the aid of a structured talk or an easy-to complete questionnaire. Discussing the problem with the patient and referring him/her to a teaching program may be the first step towards treating the depression. When considering medication, possible side effects and interactions with diabetic therapy must be taken into account. PMID- 15903057 TI - [Controversial therapeutic strategies in the treatment of type 2 diabetes mellitus]. AB - At the time of the diagnosis insulin resistance is present in the majority of type 2 diabetics. Only in those type 2 diabetics who are of normal weight at diagnosis must a disturbance of insulin secretion be assumed. These pathophysiological facts, discovered in recent years, mandate a differentiated treatment of type 2 diabetics. The primary aim must be the treatment of the insulin resistance. Insulinotropic drugs are indicated only during the further course of the disease. The use of glitazones makes possible a largely causal treatment of insulin resistance. Over the long term better glycemic control is achieved with glitazones rather than sulfonylureas. As a secondary effect, the insulin-producing beta cells are protected, and such cardiovascular risk factors as diabetic dyslipoproteinemia, hypertension or microalbuminuria are favorably influenced. PMID- 15903059 TI - [Tonsillectomy versus laser tonsillotomy for tonsillar hyperplasia in children]. AB - An appropriate indication for laser tonsillotomy is non-infectious obstructive hyperplasia in children. As a rule, these children suffer from pronounced rhonchopathy, which may be associated with episodes of apnea. Furthermore, disordered articulation and problems with eating have also been described. With increasing age hyperplastic tonsils may repeatedly become inflamed, with the result that tonsillar tissue undergoes changes that further the development of chronic tonsillitis. The indication for tonsillotomy should, however, not be applied in children older than 6 years. In children beyond this age tonsillectomy is recommended, not least due to the fact that the immunological "learning phase" is usually complete around the age of 6. PMID- 15903060 TI - [COPD patients in the physician's office]. PMID- 15903061 TI - [Indications from the anamnesis. Asthma: simply to be thought of for the first time]. PMID- 15903062 TI - [Family physician's complex performances. Spirometer to be included now?]. PMID- 15903063 TI - [Chronic inflammatory bowel diseases. Update on evidence-based therapy]. PMID- 15903064 TI - [Chronic movement pain. Early opioid therapy brings analgesia and prevention at the same time]. PMID- 15903065 TI - [Drowsiness and abdominal pain]. PMID- 15903066 TI - [Look diagnosis. Mega-gout]. PMID- 15903067 TI - [Health and sustainable development in the southern hemisphere]. PMID- 15903068 TI - [AIDS control: broadening the scope of interventions]. PMID- 15903069 TI - [Gnathostomiasis]. AB - Gnathostomiasis is an helminthic zoonotic disease endemic in Asia, mainly in Southeastern Asia, and Latin America, that has been increasingly reported in travelers returning from these areas. It is due to the consumption of raw or insufficiently cooked meat (chiken, snakes), frogs or fish, contaminated with larvae of the gender Gnathostoma. Gnathostomiasis includes a great variety of clinical manifestations caused by cutaneous and/or visceral larva migrans syndrome. We present here parasitological, epidemiological, clinical and therapeutic aspect of the disease. PMID- 15903070 TI - [Lice and lice-borne diseases in humans]. AB - Among the three lice which parasite the human being, the human body louse, Pediculus humanus humanus, is a vector of infectious diseases. It lives and multiplies in clothes and human infestation is associated with cold weather and a lack of hygiene. Three pathogenic bacteria are transmitted by the body louse: 1) Rickettsia prowazekii, the agent of epidemic typhus of which the most recent outbreak (and the largest since World War II) was observed during the civil war in Burundi; 2) Borrelia recurrentis, the agent of relapsing fever, historically responsible of massive outbreaks in Eurasia and Africa, which prevails currently in Ethiopia and neighboring countries; 3) Bartonella quintana, the agent of trench fever, bacillary angiomatosis, chronic bacteremia, endocarditis, and lymphadenopathy. Body louse infestation, associated with a decline in social and hygienic conditions provoked by civil unrest and economic instability, is reemergent worldwide. Recently, a forth human pathogen, Acinetobacter baumannii, has been associated to the body louse. PMID- 15903071 TI - [A case of imported pulmonary coccidioidomycosis]. PMID- 15903072 TI - [Avian [corrected] flu: a virus waiting in ambush]. PMID- 15903073 TI - [Meat inspection. 1. - sheep and goats]. AB - The purpose of this initial technical notice is to provide public health agents with guidelines for inspection of mutton and goat meat. Focusing on hazards related to zoonotic agents, this notice successively presents the symptoms and lesions that must be checked for to determine to approve or reject meat for human consumption. PMID- 15903074 TI - [Child abuse in Cameroon: evaluation of a training course on awareness, detection, and reporting of child abuse]. AB - The twofold purpose of this study was to promote and facilitate awareness, detection and reporting of child abuse by improving the diagnostic ability of a group of pediatric care providers working in pediatric emergency rooms and to evaluate the impact of training on awareness, detection and reporting of child abuse. This study was conducted in three phases. In the first phase data were retrospectively collected for a three-month period prior to the study (May, June, and July 1996). In the second phase a training seminar about awareness, detection, and reporting of child abuse was organized from April 29 to 30, 1997 for pediatric care providers working in the pediatric emergency rooms of several facilities in Yaounde, Cameroon. Trainees included pediatricians, general practitioners, nurses, and social workers. An oral presentation using 137-slides (visual diagnosis of physical abuse) from the American Academy of Paediatrics (1994) was used as a teaching aid. In the third phase the same data as in the first phase was prospectively collected on the same three-month period after the seminar (May, June and July 1997). As in the first phase data was collected by the same person from emergency room registers according to the WHO protocol (1994) (study of interpersonal physical abuse of children). During the first phase 39 cases of physical abuse were reported in a population 1269 consulting patients, i.e. 3.1% of the cohort. In the third phase prospective analysis of reporting following the training seminar showed that 161 cases of physical abuse were reported in a population of 1425 consulting patients, i.e., 11.3% of the cohort. The victim group (n=161) in the third phase was four-fold larger than the victim group in the first phase (n = 39). These findings suggest that the seminar achieved its main goals, i.e., to improve the diagnostic ability of the trainees and to increase detection and screening of physical abuse of children. PMID- 15903075 TI - [Prevalences of HIV, hepatitis B and hepatitis C in blood donors in the Republic of Djibouti]. AB - Screening for hepatitis B (HBV) surface antigen (Ag HBs) and for antibodies to hepatitis C (HCV) and human: immunodeficiency virus (HIV) was carried out in 9006 volunteer blood donors at the National Blood Bank in the Republic of Djibouti from 1998 to 2000. Results demonstrated the presence of Ag HBs in 934 patients (10.4%), antibodies to HCV in 21 patients (0.3%), and antibodies to HIV in 175 patients (1.9%). In comparison with neighboring countries the prevalence of HBV, HCV, and HIV infection in Djibouti was low. These findings should be used to guide preventive action against these viral infections in the Republic of Djibouti. Estimations of HIV infection (11.7%) based on modeling by the World Health Organization should be reviewed. PMID- 15903076 TI - [Tuberculosis control in Senegal: update on care services and recommendations for improvement]. AB - The goal of this study was to evaluate tuberculosis control in Senegal especially with regard to organization, quality, and availability of care services. Study was carried out from January to October 2002 within the framework of the National Turberculosis Control Program (NTCP) in 10 public hospitals and 8 private facilities including 4 doctors' offices, 2 company medical dispensaries, and 2 medical laboratories. Case observations were collected at the same time as surveying of diagnostic and therapeutic departments. In addition NTCP records for the period from 2000 to 2001 were searched. The reporting rate of new cases confirmed by positive smears is still low in Senegal, (62/100 000 inhabitants). Reporting is particularly low in rural areas where a clear-cut male predominance was observed. The cure rate also remains low (mean, 62%) mainly due to failure to complete treatment (28%). This situation contrasts with the extensive resources that have been devoted to diagnosis and treatment including field units for diagnosis (76 laboratories) and treatment (68 centers). These facilities are well integrated into the healthcare system and distributed nation-wide and provide effective care free of charge. The findings of this study demonstrate that there are serious impediments to control of tuberculosis in Senegal. Recommendations are made at various levels based on the results of problem analysis and are used to develop new management strategies aimed at improving NTCP performance indicators in Senegal. PMID- 15903077 TI - [Diagnostic and therapeutic features of tuberculosis in patients undergoing maintenance hemodialysis in Dakar]. AB - Tuberculosis in patients undergoing maintenance hemodialysis therapy presents a number of diagnostic and therapeutic challenges. This study was designed to assess the specific diagnostic and therapeutic features of dialysis-associated tuberculosis in a clinical setting. A total of 55 patients were enrolled in this retrospective study over the 5-year period from 1996 to 2000. Diagnostic techniques included questionnaire, clinical examination, chest X- ray, tuberculin skin test, Mycobacterium tuberculosis (MT) screening on biological fluids and tissue biopsy. Tuberculosis was diagnosed in 6 patients (11%). The disease occurred within the first two years after the beginning of hemodialysis. Initial signs were nonspecific, i.e., fever, weight loss, and cough. Tuberculin skin testing was negative in 5 cases. Tuberculosis was located in the lung in 2 cases, pleura in 2 cases, peritoneum in 1, and lymph node in 1. Isolation of the MT and confirmation by tissue biopsy was performed in only one case. Appropriate polychemotherapy was successful in 5 of 6 cases. Morbidity and mortality of tuberculosis in dialysis patients is closely related to early detection and treatment. Therapy is often based on strong presumptive evidence without definitive diagnosis. PMID- 15903078 TI - [Impact of canine demography on rabies transmission in N'djamena, Chad]. AB - A transversal study using the clustering technique was carried out in 600 households to estimate dog-population density in the city of N'Djamena and evaluate the knowledge of the inhabitants concerning the risk of rabies. Dog population density as estimated by negative binomial modelling was 0.03 dogs per person. The dog population estimated taking into account the extrapolated human population for 2001 was at 23 575 dogs (95% confidence interval, 14579-37921 dogs). Three-fourths of inhabitants reported that they had heard of rabies. Most respondents knew some rabies symptoms such as frothing at the mouth, restlessness, and aggressivity. In case of dog-bite injury many respondents said that they would seek care in medical facilities, e.g., health-care centers, hospitals and veterinary clinics, but some said they would also use traditional remedies, witch-doctor services, and self-medication. More than half of the respondents believed that symptomatic rabies could be cured. Because of confinement and feeding needs stray dogs are common. The canine vaccination rate is low (19%) for various reasons including limited financial resources, absence of transportation, unavailability of vaccination facilities, ignorance and negligence. Since access to health care after exposure can be difficult, mass vaccination of dogs is the recommended strategy to control rabies in cities like N'Djamena. Cooperation of all those involved in rabies control is necessary to promote the education and awareness action necessary to promote canine vaccination and reporting of exposure cases to competent authorities. PMID- 15903079 TI - [Role of vitamin deficiency in pancytopenia in Djibouti. Findings in a series of 81 consectutive patients]. AB - The purpose of this study of patients with pancytopenia in Republic of Djibouti was to identify etiologic factors and attempt to define diagnostic and therapeutic strategies adapted to local conditions. Clinical, biological and radiological assessment was performed in 81 patients hospitalized for pancytopenia. There were 56 men and 25 women. Mean hemoglobin, leukocyte and platelet rates were 56,5 +/- 22,7 g/l, 2,1 +/- 0,7.g/l and 56,2 +/- 24,7 g/l respectively. Vitamin deficiency was the most common cause of pancytopenia (49%), followed by hypersplenism (9%), HIV infection (6%) and leishmaniasis (6%). Vitamin-deficient patients had significantly more severe anemia and thrombopenia and significantly higher mean corpuscular volume than patients with pancytopenia related to other causes. Hemoglobin rate lower than 40 g/L and platelet rate lower than 35 G/L showed a positive predictive values of 90% and 100% respectively for a vitamin deficient pancytopenia. Vitamin deficiency is the most frequent etiology of pancytopenia and causes the most severe cytopenia in Djibouti. Rapid vitamin supplementation after minimal etiologic assessment including a myelogram is an effective treatment strategy for this public health problem. PMID- 15903080 TI - [How far do clandestine abortion providers go in Africa?]. AB - A 28-year-old, gravida 2, para 2 was admitted to undergo abortion in the third quarter. The procedure consisting of mechanical induction of labor using plastic catheters was performed by a male nurse. The day after placement of the catheters the patient began labor and gave uneventful birth to female infant weighing 2900 grams. The procedure undertaken to destroy the fetus led to the birth of a normal healthy child. Mechanical induction after full-term pregnancy (rarely performed in our areas) is unwarranted if pregnancy is ongoing. Due to his ignorance of the late stage of pregnancy and of the potential consequences of the procedure, the male nurse had exposed the patient to risk of severe trauma. Although the outcome was favorable for the patient, action must be taken to control intermediate level health care workers whose practices can endanger the population. PMID- 15903081 TI - [A case of meningoencephalitis caused by Acanthamoeba sp. in Dakar]. AB - Primary meningoencephalitis caused by free-living soil ameba is rare. We report the first diagnosed case of meningoencephalitis due to Acanthamoeba sp. in Senegal. The patient was a 24-year-old Senegalese woman hospitalized in the neurology department of Fann Hospital. Diagnosis was made 6 months after the onset of symptoms based mainly on headache with fever usually occurring in the evening, chills, and lumbar puncture demonstrating turbid fluid. Parasitological examination of cool cerebrospinal fluid sediment revealed the presence of free living ameba trophozoites of the Acanthamoeba genus. Species determination by culture on 1.5% agar-agar enriched with Escherichia coli failed. The patient died one month following initiation of treatment using amphotericine B. PMID- 15903082 TI - [Ivermectin]. AB - Initial clinical trials in 1980 showed that ivermectin was remarkably effective against Onchocerca volvulus. Some 25 years after more than 50 million people are treated annually with Mectizan mainly within the framework of the African Programme for Onchocerciasis Control (APOC). This success has been possible thanks to Merck Mectizan Donation Program and to distribution through a novel strategy based on the strong involvement of endemic communities. In the last few years Mectizan has been used in combination with albendazole to control lymphatic filariasis on a large-scale basis in African countries. More recently ivermectin (under the tradename Stromectol) received market approval in France for treatment of gastrointestinal strongyloidiasis and scabies. Clinical trials are under way to evaluate the activity of ivermectin on nematodes (Loa loa, Mansonella sp., intestinal nematodes, cutaneous and visceral larva migrans) and ectoparasites (Pediculus humanus capitis, Phtirius pubis, Tunga penetrans, myiases). Trials are also ongoing to explain the mechanisms underlying the severe adverse events sometimes observed in patients presenting high Loa loa microfilaraemia and to develop preventive measures. Fundamental research will provide a better understanding of the mode of action of ivermectin at the molecular and cellular level, evaluate the risk of resistance of human parasites, and to determine the extent to which ivermectin could be used in association with other agents for the treatment of nonparasitic diseases. PMID- 15903083 TI - [Renaissance of training in general surgery in Cambodia: a unique experience or reproducible model]. AB - Is the new surgical training program at the University of Phom-Penh, Cambodia a unique experience or can it serve as a model for developing countries? This report describes the encouraging first results of this didactic and hands-on surgical program. Based on their findings the authors recommend not only continuing the program in Phom-Penh but also proposing slightly modified versions to new medical universities not currently offering specialization in surgery. PMID- 15903084 TI - [Community financing for health care in Africa: mutual health insurance]. AB - Health care in sub-Saharan Africa is increasingly financed by direct payments from the population. Mutual health insurance plans are developing to ensure better risk sharing. However mutual health insurance cannot fully resolve all equity issues. The low resources available for contribution and the limited availability of care services especially in the public sector cannot guarantee the quality of care necessary for the development of mutual health insurance. National governments must not forget their responsibility especially for defining and ensuring basic services that must be accessible to all. Will mutual health insurance plans be a stepping-stone to universal health care coverage and can these plans be successfully implemented in the context of an informal economy? PMID- 15903085 TI - [Malaria control in Plasmodium falciparum resistant to multi-therapy: a field opinion]. PMID- 15903086 TI - [Shamanic healing in the age of modern medicine]. AB - This study on the therapeutic aspects of shamanic medicine differentiates the anthropologic approaches used in the 19th and early 20th century from contemporary approaches developed since the n Central to Eastern Asia. In addition to reviewing the major works on shamanic practice, this study takes into account lesser-known data on contemporary shamanic practices in Nepal. Shamanism embraces special cosmogenic beliefs. From a biological standpoint the shamanic trance like various healing techniques raises interesting questions about the involvement of certain brain regions in the development of mental factors that may generate somatic symptoms. From the therapeutic standpoint it shows the importance of managing social and psychoaffective factors. The current resurgence of shamanism could signal the revival of the disrupted ethnic identities. The shaman and medical doctor occupy symmetric, inversed positions. The shaman is a master in the management of social and psychoaffective health determinants. Until now the physician may have tended to overlook the human side of disease. PMID- 15903087 TI - [Corticosteroid therapy for allergic bronchopulmonary aspergillosis: deadly weapon]. PMID- 15903088 TI - [International Congress "Equine arboviral diseases in America and their impact in public health", Tuxtla Gutierex, Mexico, 18-20 november 2004]. PMID- 15903089 TI - [International conference on poisoning in Africa]. PMID- 15903090 TI - [Attachment and aging]. PMID- 15903091 TI - [Fear of falling: psychometric aspects of Tinetti's Falls Efficacy Scale]. AB - In this article we report on the psychometric characteristics of Tinetti's Falls Efficacy Scale. It appears that the scale is homogenous and has good internal consistency. Expected associations with age, gender, physical activity, chronic diseases and history of falls were confirmed. It is concluded that the scale is a reliable and valid instrument to measure fear of falling in performing everyday activities inside the house. PMID- 15903092 TI - [Teaching and training in geriatric medicine in the European Union]. AB - In 1993 about 20% of the population in the 15 'old' member countries of the European Union (EU) was over 60 years of age and this percentage will increase to more than 25% in 2020. These developments play a key role for the investments in education and training to meet societies needs for health care services. In 2002 about 25% of the medical students in the 'old' EU did not receive any education in geriatric medicine. A question is who will provide the services for older people in related areas, like social care, community care, acute care in the hospitals, long-term care, permanent care and care for psychiatric patients? Geriatric medicine has been recognized as an independent specialty in 8 of the 15 member countries of the 'old' EU. In all EU member states the governments are autonomous regarding all aspects of health care services, including the recognition of specialties and specialist training programmes. A two years training in internal medicine has been recommended in the EU, followed by another four years of training in geriatric medicine. The specialist training has a hospital oriented character, however, it includes also community care and other institutionalised care like nursing homes. The curriculum should contain: biological, social, psychological and medical aspects of common diseases and disturbances in older people. A problem in many EU countries is the shortage of well trained researchers and leading persons for academic positions for geriatric medicine. In a number of countries chairs at the universities remain vacant for long periods of time or even disappear. Good services in the health care for older people need a high quality curriculum and training programme. PMID- 15903093 TI - [Medication against aging? Aubrey de Grey expects it is possible]. PMID- 15903094 TI - What is the role of biomechanics in cardiac surgery? PMID- 15903095 TI - The role of hydraulic circulatory duplicators in the study of surgical operations for complex congenital cardiac diseases. PMID- 15903096 TI - A study of the Fontan circulation and ventricular energetics based on a model. PMID- 15903097 TI - In vitro experiments and in vivo implants to evaluate a new silicone-based polyurethane material for replacement of small vessels. PMID- 15903098 TI - Assessment by computational and in vitro studies of the blood flow rate through modified Blalock-Taussig shunts. PMID- 15903099 TI - Surgical experience with the modified Blalock-Taussig shunt: what we might learn from mathematical and laboratory studies? PMID- 15903100 TI - A study of mathematical modelling of the competitions of flow in the cavopulmonary anastomosis with persistent forward flow. PMID- 15903101 TI - The effect of the position of an additional systemic-to-pulmonary shunt on the fluid dynamics of the bidirectional cavo-pulmonary anastomosis. PMID- 15903102 TI - The clinical impact of antegrade pulmonary blood flow on the bidirectional cavopulmonary shunt in infants. PMID- 15903103 TI - Ten years of modelling to achieve haemodynamic optimisation of the total cavopulmonary connection. PMID- 15903104 TI - The patterns of flow in the total extracardiac cavopulmonary connection. PMID- 15903105 TI - Should we attempt functionally univentricular or partial biventricular repair for patients with complex congenital cardiac anomalies and hypoplasia of the subpulmonary ventricle? PMID- 15903106 TI - The partial cavo-pulmonary circulation with an additional source of pulmonary flow. An alternative to the total cavo-pulmonary circulation in patients with a functionally single ventricle. PMID- 15903107 TI - The Fontan operation 30 years later. PMID- 15903108 TI - Global mathematical modelling of the Norwood circulation: a multiscale approach for the study of the pulmonary and coronary arterial perfusions. PMID- 15903109 TI - Hypoplasia of the left ventricle and the physiology of the Norwood circulation. PMID- 15903110 TI - The functionally univentricular circulation in the Norwood procedure: from analysis of fluid dynamics to surgical procedures. PMID- 15903111 TI - Results of the Norwood operation for hypoplastic left heart syndrome. PMID- 15903112 TI - Experience over five years using a shunt placed between the right ventricle and the pulmonary arteries during initial reconstruction of hypoplasia of the left heart. PMID- 15903113 TI - Drug treatments for bipolar disorder: 2--maintenance, prevention and special situations. AB - In last month's issue, we discussed drug treatment for acute mood episodes (mania, hypomania, depression or mixed states) in patients with bipolar disorder. We pointed out that prescribers have to keep in mind that patients will often need treatment long term. Here we consider the role of drug therapy for maintenance and prevention in patients with bipolar disorder. We also discuss some common clinical situations that pose particular problems and dilemmas with regard to drug treatment. PMID- 15903114 TI - H. pylori eradication in NSAID-associated ulcers. AB - Helicobacter pylori infection and NSAID therapy are each closely associated with gastric and duodenal ulceration. The presence of both could have a major influence on the natural history of such disease or the efficacy of its treatment. Here we discuss the impact of eradicating H. pylori on the development and treatment of ulcers associated with NSAID therapy. PMID- 15903115 TI - What can functional neuroimaging tell the experimental psychologist? AB - I argue here that functional neuroimaging data--which I restrict to the haemodynamic techniques of fMRI and PET--can inform psychological theorizing, provided one assumes a "systematic" function-structure mapping in the brain. In this case, imaging data simply comprise another dependent variable, along with behavioural data, that can be used to test competing theories. In particular, I distinguish two types of inference: function-to-structure deduction and structure to-function induction. With the former inference, a qualitatively different pattern of activity over the brain under two experimental conditions implies at least one different function associated with changes in the independent variable. With the second type of inference, activity of the same brain region(s) under two conditions implies a common function, possibly not predicted a priori. I illustrate these inferences with imaging studies of recognition memory, short term memory, and repetition priming. I then consider in greater detail what is meant by a "systematic" function-structure mapping and argue that, particularly for structure-to-function induction, this entails a one-to-one mapping between functional and structural units, although the structural unit may be a network of interacting regions and care must be taken over the appropriate level of functional/structural abstraction. Nonetheless, the assumption of a systematic function-structure mapping is a "working hypothesis" that, in common with other scientific fields, cannot be proved on independent grounds and is probably best evaluated by the success of the enterprise as a whole. I also consider statistical issues such as the definition of a qualitative difference and methodological issues such as the relationship between imaging and behavioural data. I finish by reviewing various objections to neuroimaging, including neophrenology, functionalism, and equipotentiality, and by observing some criticisms of current practice in the imaging literature. PMID- 15903116 TI - When context hinders! Learn-test compatibility in face recognition. AB - Some theories of holistic face processing propose that parts in faces (eyes, nose, mouth, etc.) are not explicitly represented. So far, the empirical evidence has shown that whole-to-part superiority is found when wholes are learned. We substantiated this using photographic faces. More importantly, we investigated whether learning parts also reveals holistic effects. This has not been attempted before. Four experiments showed that after learning facial parts, recognition of these parts was disrupted when the part was shown in the full face. This distraction effect was strongest when perceivers were not directed to focus on a particular facial feature. Thus, it is very difficult to ignore irrelevant parts in faces. In fact, this might be the essence of holistic face processing. PMID- 15903117 TI - Exploring the role of lexical stress in lexical recognition. AB - Three cross-modal priming experiments examined the role of suprasegmental information in the processing of spoken words. All primes consisted of truncated spoken Dutch words. Recognition of visually presented word targets was facilitated by prior auditory presentation of the first two syllables of the same words as primes, but only if they were appropriately stressed (e.g., OKTOBER preceded by okTO-); inappropriate stress, compatible with another word (e.g., OKTOBER preceded by OCto-, the beginning of octopus), produced inhibition. Monosyllabic fragments (e.g., OC-) also produced facilitation when appropriately stressed; if inappropriately stressed, they produced neither facilitation nor inhibition. The bisyllabic fragments that were compatible with only one word produced facilitation to semantically associated words, but inappropriate stress caused no inhibition of associates. The results are explained within a model of spoken-word recognition involving competition between simultaneously activated phonological representations followed by activation of separate conceptual representations for strongly supported lexical candidates; at the level of the phonological representations, activation is modulated by both segmental and suprasegmental information. PMID- 15903118 TI - The effects of total sleep deprivation on the generation of random sequences of key-presses, numbers and nouns. AB - According to a recent hypothesis, executive functions should be particularly vulnerable to the effects of total sleep deprivation. Random generation is a task that taps executive functions. In three experiments we examined the effects of total sleep deprivation on random generation of key-presses, numbers, and nouns, in particular on the suppression of prepotent responses and the selection of next responses by way of applying a local-representativeness heuristic. With random key-presses suppression of prepotent responses did not suffer from lack of sleep, but it became poorer at a sufficiently high pacing rate. In contrast, suppression of prepotent responses suffered when numbers and nouns were generated. According to these findings different types of random generation tasks involve different types of inhibitory process. With only four response alternatives, but not with larger response sets, application of the local-representativeness heuristic was impaired after a night without sleep. In terms of a simple formal model, serial order representations of the preceding responses are used in selecting the next response only for the small response set, and not for larger response sets. Thus, serial-order representations are likely to suffer from loss of sleep. These findings strongly suggest that random generation involves multiple processes and that total sleep deprivation does not impair all sorts of executive functions, but only some. PMID- 15903119 TI - Generation of hypotheses in Wason's 2-4-6 task: an information theory approach. AB - We explored the "context of discovery" in Wason's 2-4-6 task, focusing on how the first hypothesis is generated. According to Oaksford and Chater (1994a) people generate hypotheses extracting "common features", or regularities, from the available triples, but their model does not explain why some regularities contribute to the hypothesis more than do other regularities. Our conjecture is that some regularities contribute to the hypothesis more than do other regularities because people estimate the amount of information in the perceived regularities and try to preserve as much information as possible in their initial hypotheses. Experiment 1, which used two initial triples, showed that the presence of high-information relational regularities in the initial triples affected the information in the initial hypotheses more than did the presence of low-information object regularities. Experiment 2 extended the results to the classic situation in which only one initial triple is given. It also suggested that amount of information is the only aspect of the structure of the triple that affects hypotheses generation. Experiment 3 confirmed the latter finding: Although relations are commonly distinguished between first-order and higher order relations, the latter being most important for generating hypotheses (Gentner, 1983), higher order relations do have an effect on Wason's 2-4-6 task only if their presence incre ases information. In the conclusion we discuss the statistical soundness of human hypotheses generation processes, and we ask an unanswered question: Amount of information explains why some regularities are preferred to others, but only within a set of "nonarbitrary" regularities; there are object regularities that are rich in information content, but are considered "arbitrary", and are not used in generating hypotheses. Which formal property can distinguish between these two sets of regularities? PMID- 15903120 TI - Categorical reasoning from multiple diagrams. AB - Syllogistic reasoning from categorical premise pairs is generally taken to be a multistep process. Quantifiers (all, no, some, some ... not) must be interpreted, representations constructed, and conclusions identified from these. Explanations of performance have been proposed in which errors may occur at any of these stages. The current paper contrasts (a) representation explanations of performance, in which errors occur because not all possible representations are constructed, and/or mistakes are made when doing so (e.g., mental models theory), and (b) conclusion identification explanations, in which errors occur even when information has been correctly and exhaustively represented, due to systematic difficulties that people may have when identifying particular conclusions, or in identifying conclusions in particular circumstances. Three experiments are reported, in which people identified valid conclusions from diagrams analogous to Euler circles, so that the first two stages of reasoning from premise pairs were effectively removed. Despite this, several phenomena associated with reasoning from premise pairs persisted, and it is suggested that whereas representation explanations may account for some of these phenomena, conclusion identification explanations, which have never previously been considered, are required for others. PMID- 15903121 TI - The importance of assessing the dose actually administered in pharmacokinetic trials. PMID- 15903122 TI - Assessment of drug-induced QT prolongation: to bin or not to bin? PMID- 15903123 TI - Clinical assessment of drug-induced QT prolongation in association with heart rate changes. AB - BACKGROUND: The formulas for heart rate (HR) correction of QT interval have been shown to overcorrect or undercorrect this interval with changes in HR. A Holter monitoring method avoiding the need for any correction formulas is proposed as a means to assess drug-induced QT interval changes. METHODS: A thorough QT study included 2 single doses of the alpha1-adrenergic receptor blocker alfuzosin, placebo, and a QT-positive control arm (moxifloxacin) in 48 healthy subjects. Bazett, Fridericia, population-specific (QTcN), and subject-specific (QTcNi) correction formulas were applied to 12-lead electrocardio-graphic recording data. QT1000 (QT at RR = 1000 ms), QT largest bin (at the largest sample size bin), and QT average (average QT of all RR bins) were obtained from Holter recordings by use of custom software to perform rate-independent QT analysis. RESULTS: The 3 Holter end points provided similar results, as follows: Moxifloxacin-induced QT prolongation was 7.0 ms (95% confidence interval [CI], 4.4-9.6 ms) for QT1000, 6.9 ms (95% CI, 4.8-9.1 ms) for QT largest bin, and 6.6 ms (95% CI, 4.6-8.6 ms) for QT average. At the therapeutic dose (10 mg), alfuzosin did not induce significant change in the QT. The 40-mg dose of alfuzosin increased HR by 3.7 beats/min and induced a small QT1000 increase of 2.9 ms (95% CI, 0.3-5.5 ms) (QTcN, +4.6 ms [95% CI, 2.1-7.0 ms]; QTcNi, +4.7 ms [95% CI, 2.2-7.1 ms]). Data corrected by "universal" correction formulas still showed rate dependency and yielded larger QTc change estimations. The Holter method was able to show the drug-induced changes in QT rate dependence. CONCLUSIONS: The direct Holter-based QT interval measurement method provides an alternative approach to measure rate independent estimates of QT interval changes during treatment. PMID- 15903124 TI - Identification and phenotype characterization of two CYP3A haplotypes causing different enzymatic capacity in fetal livers. AB - BACKGROUND: The fetal liver cytochrome P450 (CYP) 3A enzymes metabolize potentially toxic and teratogenic substrates and drugs in addition to endogenous hormones and differentiation factors. CYP3A7 is the most abundant CYP in the human liver during fetal stages and the first months of postnatal age and shows a large interindividual variability of unknown molecular basis. METHODS: A new variant gene (CUpsilonP3A7*2), which carries a mutation in exon 11 of CUpsilonP3A7 causing a T409R substitution, was identified by direct sequencing. Genotype analysis was performed by use of polymerase chain reaction followed by restriction enzyme analysis. CYP3A7.2 activity was assessed in heterologous expression systems and human fetal liver microsomes. RESULTS: The frequency of CUpsilonP3A7*2 was 8%, 17%, 28%, and 62% in white, Saudi Arabian, Chinese, and Tanzanian individuals, respectively. By use of human HEK293 cells, no significant differences in expression between CYP3A7.1 and CYP3A7.2 were found and fetal livers homozygous for CUpsilonP3A7*2 had similar or higher CYP3A7 protein contents than CUpsilonP3A7*1 livers. Kinetic studies showed that CYP3A7.2 was a functional enzyme with a significantly higher catalytic constant (kcat) as compared with CYP3A7.1 (P < .05). Interestingly, fetal livers that expressed CYP3A7.2 also expressed CYP3A5 protein, and we found a linkage disequilibrium between the CUpsilonP3A7*2 and CUpsilonP3A5*1 alleles that was subject to interethnic differences. Determination of the alprazolam 1-hydroxylation rate revealed that CYP3A5 plays a significant role in the metabolism of CYP3A substrates in the fetal liver. CONCLUSION: We have identified 2 different CYP3A phenotypes in the fetal liver--one that is the result of a CUpsilonP3A7*1/CUpsilonP3A5*3 haplotype causing CYP3A7.1 but no CYP3A5 expression and another with higher detoxification capacity, inherent in the CUpsilonP3A7*2/CUpsilonP3A5*1 haplotype, where CYP3A5 and a more active form of CYP3A7 are expressed. PMID- 15903125 TI - Identification of a dysfunctional missense single nucleotide variant of human adenylyl cyclase VI. AB - Genetic variants have been described for a range of G protein-coupled receptors (as well as for G proteins) linked to adenylyl cyclase. Furthermore, expression of these variants resulted in alterations in receptor-mediated activation of adenylyl cyclase, as well as alterations in more "downstream" effector pathways mediated by cyclic adenosine monophosphate. However, the identification of dysfunctional variants of adenylyl cyclase has been far more limited. Screening a region of the molecule that we recently demonstrated to be critical in regulation of enzyme activity, we have identified a missense single-nucleotide variant at amino acid 674 of human adenylyl cyclase isoform VI. In a population of 286 healthy white subjects, this variant has an allelic frequency of 3.1% (although 0/90 nonwhite subjects had this variant). Expression of this variant of adenylyl cyclase VI (whether expressed as the S674 human adenylyl cyclase VI [ADCY6] or the S686 ADCY6 rat analog) is characterized by a significant decrease in stimulated adenylyl cyclase activity (forskolin-stimulated activity of the S674 human ADCY6 variant was decreased to 56% +/- 6% of the activity of the A674 variant [mean +/- SEM]; n = 9; P = .004). Furthermore, subjects with the S674 variant demonstrated a significantly higher lymphocyte count (2.68 +/- 4.13 x 10(3)/mm3 versus 1.90 +/- 0.72 x 10(3)/mm3, P = .019). Paralleling this phenotype, expression of the variant was associated with attenuation of the forskolin-mediated reduction in cell growth rate to 64% +/- 5% of the effect seen with expression of the wild-type ADCY6 (n = 4; P = .001). In summary, these data demonstrate an unappreciated variant of adenylyl cyclase isoform VI that has a functional impact on both enzyme activity and cyclic adenosine monophosphate mediated regulation of cell growth. PMID- 15903126 TI - Pharmacokinetics of ximelagatran and relationship to clinical response in acute deep vein thrombosis. AB - OBJECTIVE: Our objective was to characterize the pharmacokinetics of melagatran, the active form of the oral direct thrombin inhibitor ximelagatran, and the relationship between melagatran exposure and clinical outcome in patients with acute deep vein thrombosis. METHODS: A population pharmacokinetic analysis was performed on samples from patients with deep vein thrombosis participating in a randomized dose-finding study (THRombin Inhibitor in Venous thrombo-Embolism [THRIVE I]). Patients received fixed doses of oral ximelagatran (24, 36, 48, or 60 mg twice daily) for 12 to 16 days. Thrombus size was evaluated by venography before and after treatment. Exposure-response curves were characterized for the probability of regression, no change, and progression of the thrombus extension and of having a bleeding-related event, by use of logistic regression models. RESULTS: The pharmacokinetics of melagatran (1836 samples in 264 patients) was predictable, without significant time or dose dependencies. Clearance after oral administration (population mean, 27.3 L/h) was correlated with creatinine clearance (P < 10(-6)), and volume of distribution (population mean, 176 L) was correlated with body weight (P = 2 x 10(-5)). Gender, age, or smoking did not significantly influence melagatran pharmacokinetics after the influence of renal function and body weight was accounted for. Unexplained interpatient variability values in total plasma clearance and bioavailability were 19% and 21%, respectively. The median area under the plasma melagatran concentration versus time curve across all patients and dose levels was 3.22 h x micromol/L (5th-95th percentiles, 1.35-7.69). There was no significant relationship between area under the plasma concentration versus time curve and change in thrombus extension (P = .59) or bleeding-related events (P = .77), and the estimated exposure-response curves were relatively flat. CONCLUSIONS: The pharmacokinetics of melagatran in patients with acute deep vein thrombosis was predictable after oral ximelagatran administration. Shallow exposure-response curves for efficacy and bleeding indicate that there is no need for individualized dosing or therapeutic drug monitoring in the patient population studied. PMID- 15903127 TI - Grapefruit juice ingestion significantly reduces talinolol bioavailability. AB - OBJECTIVES: Our objectives were to evaluate the effect of single and repeated grapefruit juice ingestion relative to water on the oral pharmacokinetics of the nonmetabolized and P-glycoprotein-transported drug talinolol in humans and to assess the potential impact of grapefruit juice ingestion on P-glycoprotein and intestinal uptake transporters. METHODS: The oral pharmacokinetics of 50 mg talinolol was determined with water, with 1 glass of grapefruit juice (300 mL), and after 6 days of repeated grapefruit juice ingestion (900 mL/d) in 24 healthy white volunteers. MDR1 messenger ribonucleic acid and P-glycoprotein levels were measured in duodenal biopsy specimens obtained from 3 individuals before and after ingestion of grapefruit juice. Three commonly occurring polymorphisms in the MDR1 gene were also assessed. RESULTS: A single glass of grapefruit juice decreased the talinolol area under the serum concentration-time curve (AUC), peak serum drug concentration (Cmax), and urinary excretion values to 56% (P < .001), 57% (P < .001), and 56% (P < .001), respectively, of those with water. Repeated ingestion of grapefruit juice had a similar effect (44% to 65% reduction; P < .01). Single or repeated juice ingestion did not affect renal clearance, elimination half-life, or time to reach Cmax (tmax). MDR1 messenger ribonucleic acid and P-glycoprotein levels in duodenal biopsy specimens were not affected by grapefruit juice. MDR1 genotypes (C1236T, G2677T/A, and C3435T) were not associated with altered talinolol pharmacokinetics. CONCLUSION: Because both single and repeated ingestion of grapefruit juice lowered rather than increased talinolol AUC, our findings suggest that constituents in grapefruit juice preferentially inhibited an intestinal uptake process rather than P-glycoprotein. Moreover, grapefruit juice did not alter intestinal P-glycoprotein expression. PMID- 15903128 TI - Comparison of an increased dosage regimen of rabeprazole versus a concomitant dosage regimen of famotidine with rabeprazole for nocturnal gastric acid inhibition in relation to cytochrome P450 2C19 genotypes. AB - BACKGROUND AND OBJECTIVE: A concomitant dosage regimen of a histamine 2 receptor antagonist with a proton pump inhibitor (PPI) effectively decreases the incidence of nocturnal acid breakthrough, which is one of the problems encountered when acid-related diseases are treated with a PPI alone. We compared the effectiveness of an increased dosage regimen of rabeprazole with that of a concomitant dosage regimen of rabeprazole with famotidine, relative to cytochrome P450 (CYP) 2C19 genotype status, on nocturnal acid inhibition. METHODS: Fifteen Helicobacter pylori-negative volunteers, consisting of 5 homozygous extensive metabolizers (EMs), 6 heterozygous EMs, and 4 poor metabolizers (PMs) of CYP2C19, took 20 mg rabeprazole, 40 mg rabeprazole, and 20 mg rabeprazole plus 20 mg famotidine at bedtime (at 10 PM) for 8 days. The subjects then underwent 24-hour intragastric pH monitoring on day 8. RESULTS: For the 20-mg rabeprazole, 40-mg rabeprazole, and concomitant dosage regimens, the median percent times and ranges when nocturnal intragastric pH values were lower than 4.0 were 78.8% (47.5%-98.0%), 45.3% (29.0%-52.2%), and 15.5% (0.0%-40.8%), respectively, for homozygous EMs; 51.0% (7.0%-91.6%), 41.3% (33.0%-59.0%), and 18.5% (8.4%-31.9%), respectively, for heterozygous EMs; and 4.5% (2.0%-31.2%), 9.5% (0.0%-31.1%), and 9.3% (0.0% 14.7%), respectively, for PMs. Although significant differences in acid inhibition between the different CYP2C19 genotypes were observed when rabeprazole alone was given (P = .016 for 20 mg rabeprazole and P = .023 for 40 mg rabeprazole), such differences were not observed when famotidine was concomitantly given (P = .206). CONCLUSIONS: The combination regimen of famotidine plus rabeprazole is more effective for nocturnal acid inhibition in homozygous and heterozygous EMs than the increased dosage regimen of rabeprazole. This concomitant therapy could be a rescue regimen for patients with nocturnal acid breakthrough refractory to a standard PPI therapy who are likely to be CYP2C19 EMs. PMID- 15903129 TI - Paroxetine, a cytochrome P450 2D6 inhibitor, diminishes the stereoselective O demethylation and reduces the hypoalgesic effect of tramadol. AB - OBJECTIVE: Tramadol hydrochloride (INN, tramadol) exerts its antinociceptive action through a monoaminergic effect mediated by the parent compound and an opioid effect mediated mainly by the O-demethylated metabolite (+)-M1. O demethylation is catalyzed by cytochrome P450 (CYP) 2D6. Paroxetine is a very potent inhibitor of CYP2D6. The objective of this study was to investigate the influence of paroxetine pretreatment on the biotransformation and the hypoalgesic effect of tramadol. METHODS: With and without paroxetine pretreatment (20 mg daily for 3 consecutive days), the formation of M1 and the analgesic effect of 150 mg of tramadol were studied in 16 healthy extensive metabolizers of sparteine in a randomized, double-blind, placebo-controlled, 4-way crossover study by use of experimental pain models. RESULTS: With paroxetine pretreatment, the area under the plasma concentration-time curve (AUC) of (+)- and (-)-tramadol was increased (37% [P = .001] and 32% [P = .002], respectively), and the corresponding AUCs of(+)- and (-)-M1 were decreased (67% [P = .0004] and 40% [P = .0008], respectively). (+)-M1 and (-)-M1 could be determined in all subjects throughout the study period regardless of paroxetine pretreatment. The sums of differences between postmedication and premedication values of pain measures differed between the placebo/tramadol and the placebo/placebo combination, with median values as follows: pressure pain tolerance threshold, 390 kPa (95% confidence interval [CI], 211 to 637 kPa) versus -84 kPa (95% CI, - 492 to -32 kPa) (P = .001); single sural nerve stimulation pain tolerance threshold, 25.8 mA (95% CI, 15.3 to 29.8 mA) versus 9.0 mA (95% CI, 1.5 to 14.8 mA) (P = .005); pain summation threshold, 10.7 mA (95% CI, 5.2 to 17.6 mA) versus 5.0 mA (95% CI, 2.8 to 11.2 mA) (P = .066); cold pressor pain, -4.2 cm x s (95% CI, -6.8 to -1.9 cm x s) versus -0.4 cm x s (-1.4 to 1.4 cm x s) (P = .002); and discomfort, -4.7 cm (95% CI, -10.6 to -2.8 cm) versus 0.5 cm (-0.1 to 1.4 cm) (P = .002). The sums of differences of the paroxetine/tramadol combination also differed from placebo/tramadol for some of the measures, with median values as follows: cold pressor pain, -2.2 cm x s (95% CI, -3.7 to -0.4 cm x s) (P = .036, compared with placebo/tramadol); and discomfort, -2.0 cm (95% CI, -5.6 to -1.2 cm) (P = .056). For the other measures, the hypoalgesic effect was retained on the paroxetine/tramadol combination, with median values as follows: pressure pain tolerance threshold, 389 kPa (95% CI, 141 to 715 kPa) (P = .278, compared with placebo/tramadol); single sural nerve stimulation pain tolerance threshold, 12.5 mA (95% CI, 6.2 to 28.3 mA) (P = .278); and pain summation threshold, 8.2 mA (95% CI, 4.4 to 14.6 mA) (P = .179). Paroxetine in combination with placebo showed no analgesic effect. CONCLUSIONS: It is concluded that paroxetine at a dosage of 20 mg once daily for 3 consecutive days significantly inhibits the metabolism of tramadol to its active metabolite M1 and reduces but does not abolish the hypoalgesic effect of tramadol in human experimental pain models, particularly in opioid-sensitive tests. PMID- 15903130 TI - Interpretation of opioid levels: comparison of levels during chronic pain therapy to levels from forensic autopsies. PMID- 15903131 TI - Effects of CYP2C9 polymorphisms on phenprocoumon anticoagulation status. PMID- 15903132 TI - Dose-response relationship of sublingual nitroglycerin with brachial artery dilatation and change in central and peripheral augmentation index. PMID- 15903133 TI - There is a shortage of students entering our PhD programs in audiology. PMID- 15903134 TI - Clinical genetic study of 144 patients with nonsyndromic hearing loss. AB - Hearing loss constitutes an important category of congenital defects that can be isolated or part of the phenotypic spectrum of several syndromes. A clinical genetic study was performed on a sample of 144 patients with nonsyndromic hearing loss, establishing the sex distribution, type, degree, symmetry, laterality, progression, etiology, and, when possible, inheritance pattern. PMID- 15903135 TI - Doctor of audiology (AuD) program at the University of South Florida. AB - This article describes the doctor of audiology (AuD) program at the University of South Florida (USF). The USF AuD program has been operational since the fall of 1999 and, to date, 47 students have graduated from either the postbaccalaureate track or the post-master's track. The faculty consists of 9 full-time and 8 part time adjunct members who provide a curriculum that covers the audiology scope of practice. Being located in a major metropolitan area allows for diverse clinical practicum experiences that result in students being prepared to become autonomous professional practitioners. PMID- 15903136 TI - The University of Louisville: the AuD in a medical and business environment. AB - The audiology program at the University of Louisville is located in the city's medical complex, which houses the university's health-related programs, 5 acute care hospitals, and 1 rehabilitation hospital. The audiology program has a variety of clinical sites that expose students to broad-based clinical environments, patient populations, and instructional opportunities. The presence of university faculty and staff within these clinics assures that students have consistent educational development. The expanding research program complements the clinical activities. The hospital and faculty-owned private practice locations provide important opportunities to learn the business aspects of audiology. PMID- 15903137 TI - The AuD program at the University at Buffalo. AB - The doctor of audiology (AuD) program at the University at Buffalo is summarized in this article. We begin with a description of the city of Buffalo and the University at Buffalo. This is followed by a brief history of the university and the Department of Communicative Disorders & Sciences. We then summarize the timeline and the process required to develop the AuD program. Admissions policies and prerequisites are then reviewed. We discuss the faculty active in our AuD program and present the requirements (clinical, academic, research) for the degree. The article ends with a delineation of the challenges we face, as well as a description of some of the factors that make the audiology program at the University at Buffalo unique. PMID- 15903138 TI - Genetics content in the graduate audiology curriculum: a survey of academic programs. AB - Astounding progress has been made in the identification and characterization of genes for hearing loss, which has led to an increasing role of genetics evaluation and testing in the diagnostic process for children with hearing loss. The importance of health professionals such as audiologists gaining core competencies in genetics has been recognized. The current report describes a survey of academic programs in audiology designed to determine the extent to which genetics content is included in the curriculum. Responses from 56% of existing academic programs indicate that 95% include some genetics content in their programs, with the total number of classroom hours ranging from 2 to 65. Most programs included information on basic genetic mechanisms, syndromes, and interpreting family history information, while many fewer reported covering the molecular basis of hearing loss, genetic testing, or ethical or legal issues. The results of this survey demonstrate the need to incorporate more genetics content into audiology curricula and suggest strategies for assisting audiology faculty with this process. PMID- 15903139 TI - Vestibular evoked myogenic potentials: history and overview. AB - Vestibular evoked myogenic potential (VEMP) testing is a relatively new diagnostic tool that is in the process of being investigated in patients with specific vestibular disorders. In this review, we will outline the history and provide a current review of VEMP research. Briefly, the VEMP is a biphasic response elicited by loud clicks or tone bursts recorded from the tonically contracted sternocleidomastoid muscle. Current data suggest that the VEMP is a vestibulo-collic reflex whose afferent limb arises from acoustically sensitive cells in the saccule, with signals conducted via the inferior vestibular nerve. We will review the history of the response and detail the anatomy and physiology associated with the test. We will discuss specific VEMP applications in the diagnosis of Meniere's disease, vestibular schwannoma, vestibular hypersensitivity disorders, vestibular neuritis, multiple sclerosis, and other brainstem lesions. PMID- 15903140 TI - Most comfortable and uncomfortable loudness levels: six decades of research. AB - This article critically reviews the influence of such factors as psychophysical testing method, stimulus type, and instructional set on most comfortable loudness (MCL) and uncomfortable loudness (UCL) levels. Generally, research indicates that test methods and instructions strongly affect both MCL and UCL while stimulus conditions affect them less substantially. Overall, the data suggest lower reliability for MCL than for UCL and lower reliability for pure-tone MCLs than for speech MCLs. Lower MCLs are typically obtained when measured by an ascending approach, in contrast to a descending approach. Results suggest that audiological efforts should be directed toward the development of a standardized test procedure that yields adequately reliable and valid MCLs and UCLs for routine clinical use. PMID- 15903141 TI - Effects of test order on most comfortable and uncomfortable loudness levels for speech. AB - This study examined test-order effects on most comfortable loudness (MCL) and uncomfortable loudness (UCL) levels for spondaic words in 2 groups of 30 normal hearing listeners each. For Group 1, MCL was measured first, followed by UCL. For Group 2, UCL was measured first, and then MCL. A retest was conducted for both groups. Results showed that MCL was significantly elevated for Group 2, but not for Group 1. There was no effect on UCL for either group. In a follow-up experiment, the magnitude of the test-order effect on MCL increased significantly when MCL measurements followed UCL measurements closely in time. These results argue for management of the ordering and temporal spacing of MCL and UCL testing in clinical loudness measurements. PMID- 15903142 TI - Outer and middle ear status and distortion product otoacoustic emissions in children with sickle cell disease. AB - The purpose of this study was to investigate distortion product otoacoustic emissions (DPOAEs) and outer/middle ear status in 12 African American children with normal hearing and homozygous sickle cell disease (SCD) and age-, gender-, and ear-matched African American controls. C. R. Downs, A. Stuart, & D. Holbert (2000) reported that DPOAE amplitudes were significantly larger for children with SCD. Because the integrity of the middle ear system directly influences OAE characteristics, it was felt that concurrent investigation of DPOAE amplitudes and outer/middle ear function in children with SCD was warranted. DPOAEs were evoked by 13 primary-tone pairs with f2 frequencies ranging from 1000 to 4500 Hz. Outer/middle ear status was assessed with tympanometry through indices of peak compensated static acoustic admittance, tympanometric width, tympanometric peak pressure, ear canal volume, and middle ear resonance frequency. Tympanograms were recorded with probe-tone frequencies of 226 and 678 Hz. DPOAE amplitudes were significantly larger for children with SCD (p < .05). There were no group differences in any of the middle ear indices (p > .05). These findings suggest that increased DPOAE amplitudes for children with SCD cannot be attributed to differences in outer/middle ear function as assessed with tympanometry. PMID- 15903143 TI - Exploring possible sociocultural bias on the SCAN-C. AB - Sociocultural bias on the SCAN-C (R. W. Keith, 2000) was investigated with 20 Anglo American and 20 Latino American 8-year-old children from low- and mid-high socioeconomic status (SES) backgrounds. Univariate and repeated measures analyses of variance (ANOVAs) failed to reveal any significant differences between the groups when clustered by ethnicity and SES. The Latino American participants' scores were analyzed for dialectal variations, and the ANOVA analyses were repeated using the corrected scores. No significant interactions were observed. Classification analyses revealed that 10% more Latino American children than Anglo American children fell into the borderline-to-disordered category based on SCAN-C composite scores; these classification differences were most apparent on the Filtered Words subtest (with a difference of 25%). When scores with dialectal rescoring were considered, the classification distribution for the Latino American children more closely matched that of the Anglo American children. Given the increased likelihood of Latino American children scoring in the borderline-to disordered category, caution should be used in interpreting SCAN-C results for Latino American children. Dialect scoring should be applied when Latino American children fall in the borderline-to-disordered category. PMID- 15903144 TI - Internet-based cognitive-behavioral self-help treatment of tinnitus: clinical effectiveness and predictors of outcome. AB - The aim of this investigation was to evaluate Internet-based cognitive-behavioral therapy for tinnitus in a nonrandomized clinical effectiveness study with a sample of consecutive patients referred for psychological treatment (N = 77). Results were calculated at a group level on an intention-to-treat basis and showed significant reductions of distress on the Tinnitus Reaction Questionnaire (P. H. Wilson, J. Henry, M. Bowen, & G. Haralambous, 1991), the Hospital Anxiety and Depression Scale (A. S. Zigmond & R. P. Snaith, 1983), and on the Insomnia Severity Index (C. H. Bastien, A. Vallieres, & C. M. Morin, 2001). A 3-month follow-up showed that patients remained improved. The dropout rate was 30%. Treatment compliance, external referral to the treatment, and number of earlier treatments for tinnitus were associated with positive outcome. The number of e mails between therapist and patient concerning treatment problems was associated with worse outcome. Internet-based cognitive-behavioral therapy holds some promise as a treatment modality for tinnitus. Future research should focus on further controlled evaluations of the treatment technique and evaluate the cost effectiveness compared to other forms of tinnitus treatments. PMID- 15903145 TI - Trade-offs between better hearing and better cosmetics. AB - A case study is reported of an adult bilateral cochlear implant patient who owns both a pair of ear-level and body-worn speech processors and chooses to wear them in unique configurations, knowingly compromising his auditory performance. The aim was to determine if differences in hearing could be quantified between these devices and to examine the size of these effects that would lend themselves to trading between performance and cosmetics. The patient reported wearing bilateral ear-level speech processors (programmed with the Cochlear Corporation spectral PEAK [SPEAK] coding strategy) 75% of the time for cosmetic and convenience reasons even though he "heard the best" with bilateral body-worn speech processors (programmed with the Cochlear Corporation advanced combination encoder strategy [ACE]). Speech perception and localization tests confirmed that this patient performed significantly better on monosyllabic phonemes in quiet (a difference from 60% to 75%) and localization (a total root-mean-squared-error difference from 22 degrees to 12degrees ) with bilateral body-worn speech processors and consistently rated various speech sounds as more clear than with bilateral ear-level units. There was a 2-dB difference in sentence reception threshold in noise, which was not statistically significant. These results suggest that clinicians should consider and provide options to patients when there are trade-offs to be made regarding understanding performance and cosmetics. Some individuals may choose better speech perception over cosmetics, and the ability to choose might result in greater compliance. The observations made here are relevant to hearing aid users as well. PMID- 15903146 TI - An investigation of telephone use among cochlear implant recipients. AB - The purpose of this study was to examine telephone use among cochlear implant recipients. A questionnaire was constructed and mailed to 803 adults who received a Clarion cochlear implant system manufactured by Advanced Bionics Corporation. Questionnaire recipients were implanted at least 12 months prior to receiving the questionnaire (i.e., they were implanted in 1998 or 1999). Approximately 60% (n = 478) responded, of whom 70% (n = 336) were considered "telephone users" (i.e., they answered the telephone and/or initiated calls). Telephone users were significantly younger and had significantly more daily hours of cochlear implant use than nonusers. Not surprisingly, there were differences between groups with respect to method of communication (i.e., more users employed oral communication, while more nonusers employed both oral and manual communication) and ability to understand words without lipreading (i.e., more users were able to understand). Thirty-seven percent of the telephone users were male, and 63% were female. The average age was 51.8 years (SD = 15.5). Ninety-five percent of users initiated calls to family and friends, 65% made appointments by phone, and approximately 50% asked for information about a product or service and conducted business over the phone. Over 95% of users could identify a dial tone, a busy signal, and voices. The average telephone use per week was 5.4 hr. Approximately 85% indicated that they were able to interact with strangers on the telephone within 5 months of receiving the sound processor. Approximately 30% communicated via a cellular phone for personal use. The findings of this survey suggest an increase in cochlear implant users' telephone use relative to a decade earlier. Advances in cochlear implant and telephone technologies are 2 of several factors that may contribute to the changes observed. PMID- 15903147 TI - Compromised arterial oxygen saturation in elderly asthma sufferers results in selective cognitive impairment. AB - Forty elderly patients with asthma or chronic obstructive pulmonary disorder (COPD) were compared to a comparison group of forty age-matched healthy volunteers on a range of measures of cognitive performance, and levels of arterial haemoglobin oxygen saturation recorded. Members of the patient group were found to have significantly lower oxygen saturation compared to the comparison group, and performed significantly poorer on tests of delayed word recall and serial subtractions, but not on other tasks. Correlational analysis between participants' oxygen saturation levels and test scores across the whole sample indicated significant positive relationships existed for the digit symbol substitution and serial subtractions tasks. The results are discussed in terms of cerebral oxygen delivery, glucose metabolism, age related cognitive decline, and relative task demands. PMID- 15903148 TI - Neuropsychological functioning and MMPI-2 profiles in chronic neck pain: a comparison of whiplash and non-traumatic groups. AB - We measured the neuropsychological functioning in 42 patients with chronic neck pain, 21 with a whiplash trauma and 21 without previous trauma. Subjectively, the whiplash group was more forgetful and had more concentration difficulties compared with the non-traumatic group. The neuropsychological tests did not reveal any differences between the two groups and a reference group of healthy individuals. Thus, chronic neck pain did not seem to interfere with neuropsychological functioning. The personality traits assessed with MMPI-2 in our 42 patients with chronic neck pain differed significantly from the normals on several scales. We also found that the whiplash group had more divergent test results than the non-traumatic group on the MMPI-2 test. Thus, it seems that the health status in those with chronic neck pain is closely linked to separate personality traits. It is concluded that the subjective complaints and poor performance in patients with chronic neck pain may be associated to somatization and inadequate coping, especially in chronic whiplash patients. PMID- 15903149 TI - Neuropsychological subgroups of cognitively-impaired-not-demented (CIND) individuals: delineation, reliability, and predictive validity. AB - The objectives of the present investigation were to determine whether subgroups of Cognitively-Impaired-Not-Demented (CIND) individuals with distinct neuropsychological profiles exist in two independent samples, and whether subgroup membership is related to diagnostic outcome over periods of 2 to 5 years. A series of cluster analyses was performed on ipsative factor z-scores derived from principal component analyses. Five subgroups were identified in the Base Sample (n = 461): Verbal Dysfunction, Verbal/Visuospatial Dysfunction, Memory/Verbal Dysfunction, Memory Dysfunction, and Visuospatial Dysfunction. This five-cluster solution was replicated in an independent sample of CIND individuals (n = 166). The highest rates of conversion to dementia were observed in the Memory Dysfunction and Memory/Verbal Dysfunction subgroups. The Verbal Dysfunction subgroup was most likely to show improvement in cognitive status. The cognitive heterogeneity of the CIND condition must be taken into account in future research focusing on the early identification of dementia. PMID- 15903150 TI - Neurocognitive deficits in cocaine users: a quantitative review of the evidence. AB - Studies on the neurocognitive effects of cocaine abuse are equivocal with respect to the specific types of deficits observed, although the vast majority of studies indicate that at least some deficits in certain broad functions such as attention, learning and memory, executive functions, and response speed exist. All of these studies based their results on null hypothesis statistical significance testing (NHSST). It is argued that effect size analysis, which provides information about the magnitude of difference, offers a more valid index of cognitive impairments in a population when compared to NHSST. Accordingly, the objective of the current study was to conduct an effect size analysis (or a meta analysis in cases where the same test measure was utilized in more than one study) to determine the type and the magnitude of the specific cognitive deficits found as a result of cocaine use. Effect sizes were calculated for each test variable across 15 empirical studies that met inclusion criteria. The results from 481 cocaine users and 586 healthy normal controls revealed that cocaine use had the largest effect on several measures of attention (0.40 < d < 1.10). Moderate to large effect sizes (d > 0.50) were also obtained on tests of visual memory and working memory. Minimal effect sizes (d < 0.30) were obtained on tests of verbal fluency and other language functions and sensory-perceptual functions. Tests of executive functioning produced mixed findings and were interpreted in terms of degree rather than nature of impairment. The results are consistent with findings from neuroimaging and neurochemical studies that have found cocaine use to be associated with dysfunctions in the anterior cingulate gyrus and orbitofrontal cortex; these regions are highly implicated in the mediation of attentional and executive functions, respectively. Methodological limitations of the empirical studies included in the analysis are discussed. PMID- 15903151 TI - Attention to affective pictures in closed head injury: event-related brain potentials and cardiac responses. AB - We examined whether closed head injury patients show altered patterns of selective attention to stimulus categories that naturally evoke differential responses in healthy people. Self-reported rating and electrophysiological (event related potentials [ERPs], heart rate [HR]) responses to affective pictures were studied in patients with mild head injury (n = 20; CT/MRI negative), in patients with predominantly frontal brain lesions (n = 12; CT/MRI confirmed), and in healthy controls (n = 20). Affective valence similarly modulated HR and ERP responses in all groups, but group differences occurred that were independent of picture valence. The attenuation of P3-slow wave amplitudes in the mild head injury group indicates a reduction in the engagement of attentional resources to the task. In contrast, the general enhancement of ERP amplitudes at occipital sites in the group with primarily frontal brain injury may reflect disinhibition of input at sensory receptive areas, possibly due to a deficit in top-down modulation performed by anterior control systems. PMID- 15903152 TI - The acute effects of mild traumatic brain injury on finger tapping with and without word repetition. AB - This study aimed to investigate the acute effects of mild Traumatic Brain Injury (mTBI) on the performance of a finger tapping and word repetition dual task in order to determine working memory impairment in mTBI. Sixty-four (50 male, 14 female) right-handed cases of mTBI and 26 (18 male and 8 female) right-handed cases of orthopaedic injuries were tested within 24 hours of injury. Patients with mTBI completed fewer correct taps in 10 seconds than patients with orthopaedic injuries, and female mTBI cases repeated fewer words. The size of the dual task decrement did not vary between groups. When added to a test battery including the Rapid Screen of Concussion (RSC; Comerford, Geffen, May, Medland & Geffen, 2002) and the Digit Symbol Substitution Test, finger tapping speed accounted for 1% of between groups variance and did not improve classification rates of male participants. While the addition of tapping rate did not improve the sensitivity and specificity of the RSC and DSST to mTBI in males, univariate analysis of motor performance in females indicated that dual task performance might be diagnostic. An increase in female sample size is warranted. These results confirm the view that there is a generalized slowing of processing ability following mTBI. PMID- 15903153 TI - Asymmetry in auditory and spatial attention span in normal elderly genetically at risk for Alzheimer's disease. AB - Some studies of elderly individuals with the ApoE-e4 genotype noted subtle deficits on tests of attention such as the WAIS-R Digit Span subtest, but these findings have not been consistently reported. One possible explanation for the inconsistent results could be the presence of subgroups of e4+ individuals with asymmetric cognitive profiles (i.e., significant discrepancies between verbal and visuospatial skills). Comparing genotype groups with individual, modality specific tests might obscure subtle differences between verbal and visuospatial attention in these asymmetric subgroups. In this study, we administered the WAIS R Digit Span and WMS-R Visual Memory Span subtests to 21 nondemented elderly e4+ individuals and 21 elderly e4- individuals matched on age, education, and overall cognitive ability. We hypothesized that a) the e4+ group would show a higher incidence of asymmetric cognitive profiles when comparing Digit Span/Visual Memory Span performance relative to the e4- group; and (b) an analysis of individual test performance would fail to reveal differences between the two subject groups. Although the groups' performances were comparable on the individual attention span tests, the e4+ group showed a significantly larger discrepancy between digit span and spatial span scores compared to the e4- group. These findings suggest that contrast measures of modality-specific attentional skills may be more sensitive to subtle group differences in at-risk groups, even when the groups do not differ on individual comparisons of standardized test means. The increased discrepancy between verbal and visuospatial attention may reflect the presence of "subgroups" within the ApoE-e4 group that are qualitatively similar to asymmetric subgroups commonly associated with the earliest stages of AD. PMID- 15903154 TI - Hazards in the home: using older adults' perceptions to inform warning design. AB - Older adults' perceptions of hazards associated with home product usage and beliefs about product warnings were assessed. Focus group interview data were coded along the dimensions of (a) product type; (b) hazard type; (c) hazard knowledge; (d) hazard avoidance; (e) product warning presence; and (f) product warning necessity. A survey supplemented the focus group results by gathering measures of usage patterns and usability difficulties associated with various types of household products. Older adults reported routine use of products they considered to be hazardous and identified an array of home safety concerns. Furthermore, personal experience was an important source of older adults' hazard knowledge and they described a number of hazard avoidance strategies. These data provide insight into older adults' home safety behaviour and suggest a number of approaches to improving hazard communication and warning design. PMID- 15903155 TI - Preventing injuries on horizontal ladders and track rides. AB - OBJECTIVES: This study examined the type of injury, fall heights and measures of impact attenuation of surfaces on which children fell from horizontal ladders and track rides. METHOD: All injured children who presented to two children's hospitals and received medical attention following a fall from a horizontal ladder or track ride in a public school or park during 1996--1997 were interviewed and the playground visited. RESULTS: The number of children who fell from horizontal ladders and track rides and presented to hospitals with injury was 118. Of those children, 105 were injured when they hit the ground and data were available on 102 of those playground undersurfaces. Fractures to the arm or wrist were the most common injury. The median height fallen by children was 1930 mm, 73% of injuries were from falls greater than 1800mm. In 41% of sites, the surface was deficient in impact absorbing properties for the height of the equipment. Fractures were no more likely on loose surfaces than other surfaces, such as rubber matting (p = 0.556) but more prevalent on compliant than non compliant surfaces. Relative to falls occurring on noncompliant surfaces, the odds of a fracture occurring on a compliant surface was 2.67 (95% CI 0.88-8.14). CONCLUSIONS: Modification of the height of horizontal ladders and track rides to 1800mm is preferable to removal of such equipment. The prevalence of fractures on compliant surfaces suggests that the threshold of 200g or 1000 head injury criteria (HIC) needs to be revisited, or additional test criteria added to take account of change in momentum that is not presently accounted for with either g max on HIC calculations. PMID- 15903156 TI - Child motor vehicle occupant and pedestrian casualties before and after enactment of child restraint seats legislation in Japan. AB - PROBLEM: Prevention of injuries to child passengers is a significant public health priority, as motor vehicle-related injuries remain a leading cause of death for children in Japan. The purpose of compulsory child restraint seats legislation in April 2000 was to reduce the number of child passengers killed or injured in motor vehicle crashes. METHODS: The objectives of this preliminary evaluation are to measure the effectiveness, benefits and usage of safety seats for child passengers aged 1-5 years by analysing the child casualty data for the period of 1997--2002. Population and vehicle miles travelled based injury and fatality rates were used to compare before and after legislation trends in child casualties. RESULTS: Despite overall increases in the use of child restraint seats (as observed by different national surveys), overall casualty rates in motor vehicle occupants in the 1-5 year age group did not change (fatalities and serious injuries) or even increased (minor injuries). CONCLUSIONS: Casualties among restrained children have not decreased since the law came to effect in the year 2000, perhaps because of incorrect usage of the seats. Given that exposure to crash environments is increasing, traffic safety advocates and public health community need to be aware of the importance of child restraints as a means of reducing the likelihood of injury. It is necessary to implement effective community-based child safety seat campaigns to disseminate the information on appropriate restraint use and to increase efforts to enforce the existing legislation. PMID- 15903157 TI - Community-based programmes to prevent pedestrian injuries in children 0-14 years: a systematic review. AB - BACKGROUND: Community-based models for injury prevention have become an accepted part of the overall injury control strategy. This systematic review of the scientific literature examines the evidence for their effectiveness in reducing pedestrian injury in children 0-14 years of age. METHODS: A comprehensive search of the literature was performed using the following study selection criteria: community-based intervention study; target population was children under 14 years; outcome measure is either pedestrian injury rates or observed child pedestrian or vehicle driver behaviour; and use of a community control or an historical control in the study design. Quality assessment and data abstraction was guided by a standardized procedure and performed independently by two authors. Data synthesis was in tabular and text form with meta-analysis not being possible due to the discrepancy in methods and measures between the studies. RESULTS: The review found four studies that met all the inclusion criteria. The three studies using injury as their outcome measure found a 12%, 45% and 54% reduction for all childhood injuries with the fourth showing improved traffic control at child pedestrian sites (9% reduction in traffic flow) and sustainable community safety promotion activity. CONCLUSION: There is a paucity of research studies in the literature from which evidence regarding the effectiveness of community-based programmes for the prevention of pedestrian injury in children can be drawn. However, the hypothesis that community-based interventions are effective in reducing the incidence of childhood pedestrian injury would appear to be supported, with the degree of success being cumulative depending on the complexity of individual strategies employed. PMID- 15903158 TI - The causes and control of loader- and truck-related fatalities in surface mining operations. AB - At surface mining operations throughout the world, loaders and trucks are a primary means of material loading and haulage. As the size, use and technological complexity of these units have increased, so has the concern regarding loader and truck safety. The severity and number of accidents involving loaders and trucks is higher when compared to all other mining accident types. In this paper, an analysis of loader and truck-related fatalities over the last 8 years is performed, the fatality categories and causes of accidents are established and control strategies are discussed and evaluated in an effort to increase hazard awareness by emphasizing safe loading, hauling and maintenance practices, as well as the value of traditional and innovative miner training programmes. PMID- 15903159 TI - Role of physicians in preventing accidents in the home involving children under 15 years in the French-speaking community of Belgium. AB - OBJECTIVES: To measure GPs' and paediatricians' expectations, attitudes, priorities and demands in the area of promoting safety and preventing accidents in the home involving children under 15 years of age. METHODS: A phone survey of a random sample of GPs and paediatricians in the French-speaking community of Belgium (Wallonia and Brussels) conducted in the course of September and October 2000. RESULTS: Close to two-thirds of the physicians surveyed stated that they had been contacted at least once in the 2 weeks preceding the survey to handle an injury. Of the physicians, 80% mention accident prevention after a childhood injury, but only 46% mention it during a routine consultation (whatever the reason of the latter may be). The main obstacles to mentioning prevention during routine consultation put forward by the interviewees are: 1) reasons for consultations that do not permit such an approach (79%); 2) the fact that injuries are not priorities for them (66%); 3) the lack of interesting materials to provide the subject with documentation (63%); 4) the unsuitability of the place where the contact occurs for such discussion, given the time required (56%); 5) insufficient information on the subject (41%); and 6) the patient's lack of interest (39%). An overwhelming majority (98%) nevertheless feel that they have a role to play in preventing children's accidents in the home, with 72.5% seeing this as informative (recommendation on safety rules). More than two thirds of the respondents feel that they have enough requisite information to play such a role. Those who declare that they have not enough information (34%) proposed some priority subjects about which they would like to find information or additional information to be more effective in preventing accidents. The information needs mentioned most frequently were a systematic review of the risks, of the effective prevention strategies and epidemiological data. CONCLUSIONS: The present study clearly reveals the interest of physicians for accident prevention and puts forward the current obstacles to offering prevention advice during routine consultation. The obstacles mentioned are fairly comparable to those mentioned in other studies, namely, because the reason for the visit does not give such an opening, the lack of appropriate materials and information, the lack of time, the patient's lack of interest, the fact that the issue is not a priority, etc. The problem of lack of priority for certain practitioners underlines the importance of making accident mortality and morbidity statistics available to doctors in order to improve their perception of the magnitude of the problem. The lack of interesting education materials and useful information seems to be a major reason for their failure to carry out such prevention work successfully. These factors should thus be taken into account when developing any policy and/or programme aimed at 'using' GPs and paediatricians in the prevention strategies that are adopted. PMID- 15903160 TI - World Health Assembly resolutions on violence and injury prevention: new opportunities for national action. PMID- 15903161 TI - Alcohol use and trauma in Cape Town, Durban and Port Elizabeth, South Africa: 1999--2001. AB - OBJECTIVE: To assess acute alcohol intoxication among patients presenting with recent injuries at trauma units in Cape Town, Port Elizabeth and Durban from 1999 to 2001. DESIGN: Cross-sectional surveys were conducted during a four-week period in each of the above sites in 1999, 2000 and 2001. The concept of an 'idealised week' was used to render representative samples. Breath-alcohol concentrations were assessed in a total of 1900 patients using a Lion SD2 alcolmeter. RESULTS: Over half of all the patients experienced violent injuries. Across sites and for each respective year of the survey, between 35.8% and 78.9% of patients tested positive for alcohol. Between 16.5% and 67.0% had a breath-alcohol concentration greater than or equal to 0.05g/100ml. Port Elizabeth consistently had the highest proportion of patients testing positive for alcohol. Patients injured as a result of violence were more likely to test positive for alcohol than patient who sustained road traffic or other unintentional injuries. CONCLUSIONS: Alcohol involvement among trauma patients remained consistently high for each of the three study periods. Efforts to combat the abuse of alcohol would appear to paramount in reducing the burden of injuries on health care services. PMID- 15903162 TI - Using an electronic medical record system to describe injury epidemiology and health care utilization at an inner-city hospital in Indiana. AB - Injuries are a major public health problem worldwide. In the USA, injuries cause 146, 400 deaths annually, with 31 million non-fatal injury visits to emergency departments (EDs). EDs thus represent an important source of injury data. The primary objective of the current study was to describe the epidemiology of injury related ED visits and assess injury-related utilization of health care resources in an inner-city hospital in Indiana, using data stored in a computerized medical record system. It involved a retrospective review of the records for injury visits to EDs and injury admissions over a 3-year period. The variables extracted and analysed included patients' demographics, external cause of injury, diagnosis, length of stay, ED and hospital charges. A total of 60,470 injury related ED visits were made, the majority of patients were male (61.6%), uninsured (63.1%), treated in ED and discharged (98.4%). The leading causes of injury were falls (18.8%), motor vehicle crashes (18.4%), assaults (17.6%), being struck (11.2%) and overexertion (10.6). Firearms caused most injury deaths (32.4%; n = 314); motor vehicle crashes were the leading cause of hospitalization (26.6%; n = 642) and also the most expensive to treat as inpatients (mean charge $19,190). The mean charge per patient treated and discharged was $150 compared to $11,116 for patients admitted. These findings demonstrate the value of computerized medical records in capturing and storing E-coded injury data. The system generates data that can be used for epidemiological surveillance and injury prevention at the local level, and for assessment of impact of specific injuries on health care resources. PMID- 15903163 TI - Skiing and snowboarding injuries and their impact on the emergency care system in South Tyrol: a restrospective analysis for the winter season 2001--2002. AB - OBJECTIVES: To evaluate the incidence and the pattern of skiing and snowboarding injuries in South Tyrol and their impact on the emergency medical system in the winter season 2001--2002 in an attempt to rationalize and improve the emergency care and assist in prevention strategies. METHODS: All medical records of patients referred to our emergency department (ED) that sustained a skiing or snow-boarding injury during the study period were retrospectively reviewed. Age, sex, local or non-local residency, type of injury, data and time of accident, type of transport to the hospital, hospital admission or ED discharge, Injury Severity Score, outcome (including mortality) were evaluated. On site mortality data were obtained from the emergency call-center registry. Ski resorts utilization was estimated from the data published by the Regional Office of Cable Transport. RESULTS: For the period analyzed approximately 2,500,000 skier and snowboarder days were recorded in the whole region of which about 500,000 were attributed to the four nearby ski resorts that refer to our hospital. Of the 1087 patients, 794 were skiers and 294 were snowboarders. Snow-boarders were younger than skiers (mean age 20 and 36 respectively, p = 0.001). Females were equally represented in the two groups. Male patients, children, senior skiers and non local residents suffered from more severe injuries than their corresponding classes (p < 0.01, p = 0.002, p = 0.02, p = 0.000 respectively). Critical injuries (ISS > or = 25) were homogeneously spread in the groups, with the exception of the non-local resident patients that showed a higher incidence (p < 0.02). No difference in severity was found between skiers and snowboarders. The incidence was 2.05 per 1,000 skier-days. Mortality rate was 1.6 per 1,000,000 skier-days. The pattern of injury was different: snowboarders showed more forearm and wrist trauma and skiers more lower extremity injuries. 208 patients were hospitalized and the mean length of stay was 4.5 days. Head trauma and fractures were the most common diagnosis of admission. The lack of field triage led to 12% of unjustified helicopter transfer and 9.6% of avoidable ambulance transport. CONCLUSIONS: Incidence, pattern of injuries and mortality from skiing and snowboarding accidents in South Tyrol resemble those reported in other part of the world. Nevertheless, strategies for prevention are needed. The routine use of helmets should be enforced by law. Dangerous behaviors should be prosecuted. Skiers and snowboarders should be made aware that skiing beyond their technical ability can be life-threatening. PMID- 15903164 TI - School backpack weights: a survey of students in Ghana, Guatemala and the USA. AB - Cross-sectional studies were conducted in conveniently selected schools in Ghana, Guatemala, and the USA to quantify and compare the percentage of body weight carried by students in their backpacks as well as assess parental knowledge of the backpack weights and contents. Mean backpack weights varied significantly by country and increased significantly with increasing student grade. In the USA, girls significantly carried heavier backpacks (p <.0001). The backpack loads represented an average of 7.7% (95% CI = 7.4-8.0), 9.7% (95% CI = 9.2-10.2), and 9.4% (95% CI = 9.1-9.8) of student body weights in Ghana, Guatemala, and the USA, respectively. No student in Ghana carried a wheeled backpack in contrast to 6% and 2.8%, respectively, in Guatemala and the USA. In Ghana, 4.1 and 73.6% of parents were reported to be aware of their children's backpack weights and contents, respectively, compared to 21.9 and 57.5% in Guatemala, and 7.7 and 55.3% in the USA. Backpack loads respresent a significant percentage of the body weights of students in all three countries. The vast majority of parents are unaware of their child's backpack weight. PMID- 15903165 TI - Patient payment for emergency medical services treatment and its impact on injury control: an Alberta case. AB - A province-wide qualitative study on emergency medical services (EMS) in Alberta was initiated to better understand the extent to which patient billing influences EMS performance. After completing lengthy interviews with 108 EMS-related subjects it was learned through qualitative research methods that billing patients for EMS treatment (ambulatory care) has subtle or determined influences on EMS providers' emergency actions. Furthermore the act of billing patients can and does have an impact on patient care. It often leads to patients and/or their representatives refusing treatment and transportation. Testimony indicates that the final result can be and is troublesome to, patient outcomes. The findings support the generalization that in Alberta the patient's responsibility to pay for needed EMS treatment can be a major obstacle for inclusive injury control. PMID- 15903166 TI - International collaborative effort on injury statistics: 10-year review. AB - International comparisons of injury data may be useful for examining differences in risk and for suggesting potential interventions or hypotheses for future studies. However important issues to be considered in conducting comparisons are related to both the underlying quality of the data and how the data is collected. The International Collaborative Effort (ICE) on Injury Statistics grew out of concerns over the comparability of international injury data. This article outlines the history behind the development of the injury ICE, brief descriptions of current and past projects, collaborations, and reflections on the value of collaboration. PMID- 15903167 TI - Guidelines for conducting community surveys on injuries and violence. PMID- 15903168 TI - Rehabilitation of edentulous mandibles by means of turned Branemark System implants after one-stage surgery: a 1-year retrospective study of 152 patients. AB - BACKGROUND: Rehabilitation of the edentulous mandible with oral implants is today predominantly executed with one-stage surgery and early or immediate loading. It is generally claimed that the outcome is similar to that of the classic two-stage technique. PURPOSE: The aim of the present investigation was to retrospectively evaluate the 1-year results of one-stage surgery and early loading performed in edentulous mandibles in a large group of patients. The outcome was compared with that of a study, from the same clinic (control), that used the two-stage surgical technique in edentulous mandibles and whose data were well controlled. MATERIALS AND METHODS: The study included 152 individuals with 750 turned Branemark System implants of various designs placed in edentulous mandibles by means of one-stage surgery. The prosthetic procedure was commenced at a mean of 13 days after the surgical intervention. Intraoral apical radiography was performed at the time of prosthesis placement and at the 1-year annual checkup. Comparison of failure rates between the test and the control groups was made by means of the chi-square test. RESULTS: A total of 18 implants in 12 patients in the study group were found to be mobile up to and including the first annual checkup, equivalent to a 1-year implant cumulative survival rate (CSR) of 97.5%. The corresponding CSR for the control group was 99.7%. Differences between the two groups in regard to implant survival reached significant levels when analyzed with the chi-square test (p < .05). No such significant difference was seen on the patient level (p > .05). Because of implant failures one prosthesis in the study group was remade. The mean marginal bone resorption during the first year of function was 0.4 mm in both groups. CONCLUSIONS: The present investigation showed a high but (compared with the classic two-stage technique) somewhat lower CSR after 1 year for the one stage technique. More prosthetic adjustments due to implant failures were observed in the study group, and the results emphasize the need for large study samples in order to statistically verify small differences between various treatment techniques. PMID- 15903169 TI - Functional restoration of implants on the day of surgical placement in the fully edentulous mandible: a case series. AB - PURPOSE: The purpose of this article is to report the clinical experience and outcome of a study of the functional rehabilitation of 16 completely edentulous mandibles with immediately loaded cross-arch screw-retained hybrid prostheses at the University of Southern California. MATERIALS AND METHODS: After signing informed consent forms 16 patients (9 male, 7 female) aged 47 to 84 years (mean age, 62.6+/-11.6 years) received 90 Branemark System Mk III dental implants (Nobel Biocare USA, Yorba Linda, CA, USA). Stability and radiographs of the dental implants were evaluated at the time of surgery, at 3 months, at 1 year, and at 3 years post loading. RESULTS: Three implants failed to meet the criteria of success, bringing the cumulative success rate to 96.6%, with a 100% prosthetic success rate at 3 years. Thirty-nine (43.3%) of the dental implants placed were 15 mm in length. Seventy-seven (85.5%) of the dental implants were placed in high density bone. At 3 years post loading, the average bone loss was -1.2+/-0.1 mm. CONCLUSION: Within the limitations of this study, restoration of implants by unreinforced hybrid prostheses at the time of placement provided satisfactory results. The outcome was stable at 3 years post restoration. Mandibular rehabilitation by functional loading of the implants on the day of the insertion requires the comprehension and proper application of surgical and restorative principles. PMID- 15903170 TI - Bone formation at titanium implants prepared with iso- and anisotropic surfaces of similar roughness: an in vivo study. AB - BACKGROUND: Implant surface topography influences the bone response after implantation. However, the importance of surface orientation is not known. PURPOSE: The aim of this study was to investigate the bone tissue response and the stability of titanium implants prepared with isotropic and anisotropic surfaces of similar roughness. MATERIALS AND METHODS: A total of 18 implants were divided into two groups and were inserted into the femurs of nine rabbits for 12 weeks. Confocal laser scanning microscopy was used for the topographic description to verify that the two different surfaces were modified as intended. The stability of the implants was recorded by resonance frequency (RF) measurements at insertion and at time of removal, after which the implants were evaluated histomorphometrically. RESULTS: RF measurements showed that implant stability increased with time. However, there was no significant difference between the two different surface modifications at insertion and after 12 weeks. The histomorphometric comparison revealed no statistically significant differences in regard to either bone-to-metal contact or bone area inside the threads. CONCLUSION: Titanium implants prepared with isotropic and anisotropic surfaces of similar roughness integrate similarly to bone during the 3 months after implantation. PMID- 15903171 TI - A prospective study of immediate functional loading, following the Teeth in a Day protocol: a case series of 55 consecutive edentulous maxillas. AB - BACKGROUND: Immediate loading of dental implants is increasingly gaining recognition as a viable option for both patient and clinician. PURPOSE: The aim of this study was to evaluate the results of 55 patients in a clinical investigation of immediate functional loading of Branemark System implants (Nobel Biocare USA, Yorba Linda, CA) in edentulous maxillas. Its further purpose is to suggest a reliable and evidence-based protocol for immediate implant loading of full-arch prostheses in the maxilla. MATERIALS AND METHODS: A total of 552 Branemark System implants were placed in immediate extraction or healed sites; a mean number of 10 implants were placed per patient. The healthy subjects in need of full-arch maxillary implant reconstruction were treated between December 1999 and February 2004; 522 of the 552 implants were immediately loaded with screw retained all-acrylic fixed prostheses at the time of surgery. Approximately 4 to 6 months later, the 30 submerged implants were uncovered, and a definitive metal reinforced prosthesis was delivered to each patient. RESULTS: The immediately loaded implant cumulative survival rate was 99.0% for these patients. The prosthesis survival rate was 100%. CONCLUSIONS: The results of this prospective study of full-arch maxillary immediate loading suggests that this protocol is suitable for most patients in need of full maxillary implant reconstruction. The protocol, as shown in this study, is highly successful in providing a lasting state of osseointegration as the foundation for long-term stability of screw retained fixed prostheses. PMID- 15903172 TI - Influence of different prophylactic antibiotic regimens on implant survival rate: a retrospective clinical study. AB - BACKGROUND: The routine use of antibiotics in oral implant treatment seems to be widespread. The pre- or postoperative use of antibiotics in conjunction with implant surgery and its correlation with failure and success rates are poorly documented in the literature. The debate regarding overprescription of antibiotics raises the need for a critical evaluation of proper antibiotic coverage in association with implant treatment. PURPOSE: The purpose of this study was to compare the implant survival rate following a 1-day single-dose preoperative antibiotic regimen with that following a 1-week postoperative antibiotic protocol. MATERIALS AND METHODS: The study included 868 consecutively treated patients. A total of 3,021 implants were placed. The population was split into two categories, either receiving a 1-day single-dose administration only, or a 1-week postoperative administration of antibiotics. Healing was evaluated at second-stage surgery (6 months for the upper jaw, 3 months for the lower jaw). Failure was defined as removal of implants because of non-osseointegration. Statistical analyses were performed with analysis of variance and the Scheffe test, with a significance level of 5% for comparison of data. RESULTS: No significant differences with regard to complications and implant survival were found in the study. CONCLUSION: Based on the present data, a more restrictive regimen consisting of a 1-day dose of prophylactic antibiotic in conjunction with routine implant procedures is recommended. PMID- 15903173 TI - A 10-year follow-up study of titanium dioxide-blasted implants. AB - BACKGROUND: Dental implants with moderately rough surfaces are commonly used in the treatment of edentulous patients. However, long-term data on survival rates and marginal bone conditions are lacking. PURPOSE: This prospective study evaluated the cumulative survival rate of the TiOblast implant (Astra Tech AB, Molndal, Sweden) after 10 years of prosthetic loading. MATERIALS AND METHODS: A total of 199 TiOblast implants were placed in 36 consecutive edentulous patients (23 males and 13 females). All patients were treated at one clinic and by the same team. The patients were edentulous in either the maxilla (n = 16) or the mandible (n = 20). The average age of the patients at the start of the trial was 64 years (range, 59-82 years). Of the 199 implants inserted 108 were in the mandible and 91 were in the maxilla. Clinical evaluations were undertaken after completion of the prosthetic superstructure (baseline) and after 6 months, 1 year, 3 years, 5 years, 7 years, and 10 years. Mean marginal bone level was evaluated for the first 100 placed implants for up to 7 years. RESULTS: Six implants failed during the study (3 in the mandible and 3 in the maxilla). All failures occurred within the first year, giving a cumulative survival rate of 96.9% (96.6 % in the maxilla and 97.2 % in the mandible) after 10 years of follow up. The survival rate for the superstructures was 100%. The mean marginal bone level in the measured sample was 0.2 mm (standard deviation [SD], 0.31) below the reference point at baseline, 0.28 mm (SD, 0.20) and 1.27 mm (SD, 1.15) below the same point 7 years later (mean, 0.15 mm per year). CONCLUSION: This study showed that titanium dioxide-blasted implants offer predictable long-term results as supports for fixed prostheses in both the maxilla and mandible. PMID- 15903174 TI - Photogrammetry and conventional impressions for recording implant positions: a comparative laboratory study. AB - BACKGROUND: The development of digitized techniques for manufacturing implant frameworks has made possible alternative "impression" techniques for recording implant positions. PURPOSE: The objective of the present study was to test the precision and accuracy of a three-dimensional photogrammetric technique to record implant positions in vitro and to compare casts made with this technique with conventional casts fabricated with two conventional impression techniques. MATERIALS AND METHODS: Twenty casts were fabricated from 10 polyether (Impregum, ESPE Dental AG, Seefeld, Germany) impressions and 10 plaster (Kuhns Abdrucksgips, Ernst Hirnischs GmbH, Goslar, Germany) impressions of one master model. The casts were measured in a coordinate measuring machine (Zeiss Prismo VAST, Oberkochen, Germany) and compared with the master model. Six separate three-dimensional photographs of the master model were taken with a special camera. After the photographs were measured with an analytic plotter, results were analyzed and compared to the coordinates of the original model and casts. RESULTS: A systematic pattern of distortion in the x-axis was found for the two impression techniques. Expansion of the implant arch at the terminal implants (p < .01) averaged 22 microm and 94 microm on photographs and plaster casts, respectively. Polyether casts contracted an average of 52 microm when compared with the master (p < .01). In absolute figures, photogrammetry and the polyether technique reproduced the x-axis and three-dimensional parameters more accurately than the plaster technique did when cylinder center point distortion was compared (p < .05 to p < .001). However, angular cylinder distortion in absolute figures was greater with the photographic technique than with either of the impression techniques (p < .05-p < .001). CONCLUSION: Photogrammetry is a valid option for recording implant positions and has a precision comparable to that of conventional impression techniques. At present, however, it is limited to framework fabrication techniques that are based on digital platforms. PMID- 15903175 TI - Clinical consequences of IL-1 genotype on early implant failures in patients under periodontal maintenance. AB - BACKGROUND: Implant failure and biologic complications such as periimplantitis are not completely avoidable. Are there any genetic and microbiologic parameters that could be used to identify patients at risk for implant failure, preferably prior to treatment? This would result in improvement of the diagnostics, treatment decision, and risk assessment. PURPOSE: The aims of this retrospective study were to describe (1) the absolute failure rate of Branemark System implants (Nobel Biocare AB, Goteborg, Sweden) consecutively installed over a 10-year period in partially edentulous patients treated for periodontal disease prior to implant treatment and under regular professional maintenance, (2) the rate of interleukin-1 (IL-1) polymorphism in those patients who experienced at least one implant failure during the first year of function, and (3) the prevalence of periodontal pathogens in dental and periimplant sites with and without signs of inflammation. MATERIAL AND METHODS: Of 766 patients, 81 encountered at least one implant failure; 22 patients were clinically examined and were tested genetically for IL-1 genotypes. The presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella nigrescens was analyzed. RESULTS: The absolute implant survival rate for the whole population was 95.32%; 10.57% of the patients encountered an implant loss. Implant loss in the examined group (n = 22) was 32 of 106 (30.1%); 10 (45%) of the 22 patients were smokers, and 6 (27%) of the 22 patients were IL-1 genotype positive. Patients positive for IL-1 genotype were not more prone to implant loss; however, a significant synergistic effect with smoking was demonstrated. Between patients who were IL-1 genotype positive and those who were IL-1 genotype negative, the differences in regard to bleeding on probing or periodontal pathogens did not reach statistical significance. CONCLUSION: The overall implant failure rate in a population treated and maintained for periodontal disease is similar to that of healthy subjects. A synergistic effect found between smoking and a positive IL-1 genotype resulted in a significantly higher implant loss. This indicates that further research with a larger patient group should focus on multifactorial analysis for adequate risk assessment. PMID- 15903176 TI - Lead content and exposure from children's and adult's jewelry products. PMID- 15903177 TI - Analysis of nicotine in California air samples from XAD-4 resin. PMID- 15903178 TI - Effect of air flow on emission of smoldering incense. PMID- 15903179 TI - Chromium in some herbal drugs. AB - The use and demand of herbal drug is increasing day by day in both developing and developed countries due to the growing recognition that these are natural products having no or little side effects and can easily be available at affordable prices. Sometimes these are the only source of health care available to poor people particularly in South East Asia where more than 80% population depend on herbal drugs for their health care needs. Thus the quality and safety of herbal products is very important in order to have desired therapeutic efficacy. But in today's polluted environment with increasing automobile consumption and industrialization, WHO is emphasizing emphatically that without heavy metal assessment the herbal drug should not be used. The concentration of heavy metals including Cr is increasing in the environment and many hazardous effects are caused in the inhabitants of that environment (Prasad and Hagemeyer, 1999; Nriagu and Pacyna, 1998). For example, in human beings, due to Cr numbers of disorders occur like damage to liver, kidney, respiratory and nerve tissues. Besides enhancing risk of human lung cancer due to automobile exhaust and even irritation of skin have also been reported by excess of Cr (Anonymous, 1998; Anonymous 1988). Therefore it is mandatory to assess the Cr concentration in herbal drugs before use. Although some work has been carried out by Rai et al., 2001 a, b; Fuh et al., 2003; Al Ajasa et al., 2004 and Ernst 2002 but it is not enough and lot of work is required in this field. PMID- 15903180 TI - Adsorption of cadmium and lead by various cereals from Korea. PMID- 15903181 TI - Cadmium in feathers of adults and blood of nestlings of three raptor species from a nonpolluted Mediterranean forest, southeastern Spain. PMID- 15903182 TI - Effects of the contaminants from Turgutlu-Urganli thermomineral waters on cold ground and surface waters. PMID- 15903183 TI - Arsenic concentrations in the surface sediments of the Magdalena-Almejas Lagoon Complex, Baja California Peninsula, Mexico. PMID- 15903184 TI - Preliminary analysis of the impact of cold mix asphalt concretes on air and water quality. PMID- 15903185 TI - Nontarget deposition of methiocarb applied to a foliage plant staging area. PMID- 15903186 TI - Iron, cadmium, chromium, copper, cobalt, lead, and zinc distribution in the suspended particulate matter of the tropical Marabasco River and its estuary, Colima, Mexico. PMID- 15903187 TI - Heavy metals in tissue samples of Finnish moose, Alces alces. PMID- 15903188 TI - Persistent organochlorine pesticide residues in fish from Guanting Reservoir, People's Republic of China. PMID- 15903189 TI - Variation of lead, cadmium, copper, and zinc in aquatic macrophytes from the Seyhan River, Adana, Turkey. PMID- 15903190 TI - Chiral signatures of alpha-hexachlorocyclohexane in surface water, porewater, and sediment system. PMID- 15903191 TI - Acute toxicity profile of 1-butyl-3-methylimidazolium chloride. PMID- 15903192 TI - Genotoxic evaluation of sodium fluoride and sodium perborate in mouse bone marrow cells. PMID- 15903193 TI - Phytochemical biomarkers in eight aquatic plant species exposed to metsulfuron methyl. PMID- 15903194 TI - Characterization of the mangrove oyster, Crassostrea rhizophorae, as a biomonitor for mercury in tropical estuarine systems, northeast Brazil. PMID- 15903195 TI - Evaluation of genotoxicity of combined pollution by cadmium and atrazine. PMID- 15903196 TI - Effects on metallothionein levels and other stress defences in Senegal sole larvae exposed to cadmium. PMID- 15903197 TI - Effects of aqueous extracts from soils in nonsanitary waste landfills on germination and seedling growth of some herbaceous species. PMID- 15903198 TI - Acute toxicity of synthetic pyrethroids to Indian major carp, Catla catla L. PMID- 15903199 TI - Effects of acid precipitation and aluminum on carbohydrate metabolism in mycorrhizae of Pinus massioniana. PMID- 15903200 TI - Endoscopic electrosurgical management of posterior epistaxis: shifting paradigm. AB - Recent literature has embraced the use of electrosurgery, sphenopalatine vascular clipping, and endoscopy in posterior epistaxis. With the advent of endoscopy, the surgical treatment of posterior epistaxis has shifted from internal maxillary ligation to endoscopic sphenopalatine artery control. This article introduces an endoscopic electrosurgical approach for patients suffering from idiopathic posterior epistaxis that combines one or more of the following methods: endoscopic selective branch cauterization, endoscopic sphenopalatine artery cautery, and endoscopic posterior nasal cauterization. This approach is currently used in a hospital-based community otolaryngology practice and is presented from a Canadian health care perspective (Quebec). This article presents the experience in a series of 17 patients treated during a 35-month period and prospectively followed and discusses the surgical technique, patients' outcome, and the implications of such a practice. PMID- 15903201 TI - High-resolution X-ray computed tomographic scanning of the human stapes footplate. AB - OBJECTIVES: The mechanical behaviour of the footplate and its annular ligament depends critically on their shape and orientation in the oval window, but accurate measurements have been difficult to make owing to their small size. Our aims are to visualize the footplate at high resolution and understand its dynamics. METHODS: The human cadaver stapes footplate was dissected, and very high-resolution x-ray computed tomographic (CT) scans, with voxel sizes from 4 to 8 pm, were performed. Locally developed software was used to reconstruct the images. RESULTS: The data permit us to explore minor details of shape and orientation. The footplate looks like a footprint, and the annular ligament has variable thickness, with a cleft (groove) in its anterior attachment to the oval window. The CT data also permit us to create a three-dimensional finite-element model that can simulate footplate motion. CONCLUSIONS: The results obtained lead to further understanding of the mechanical behaviour of the footplate and the annular ligament. PMID- 15903202 TI - Outcomes for children with submucous cleft palate and velopharyngeal insufficiency. AB - OBJECTIVE: To review the outcomes of children with submucous cleft palate who also have velopharyngeal insufficiency (VPI). METHODS: A retrospective chart review was carried out at a tertiary care academic centre of all patients who had VPI with a submucous cleft palate. The University of Iowa Cleft Palate registry parameters encompassing nasality (hyper- and hyponasality) were compared pre- and postoperatively. RESULTS: Preliminary results demonstrate a significant, stable improvement in children who underwent palatal surgery for VPI. CONCLUSION: Positive outcomes in the treatment of VPI in the submucous cleft palate population were demonstrated with a combined approach of speech therapy and palatal surgery. PMID- 15903203 TI - Combined tuning fork tests in hearing loss: explorative clinical study of the patterns. AB - OBJECTIVE: To combine the tuning fork tests of Rinne, Weber, and absolute bone conduction, explore their clinical patterns, and study their combined specificity, sensitivity, and validity in assessing the hearing status of an individual. DESIGN: A cross-sectional study of the tests of Rinne, Weber, and absolute bone conduction was done on 317 adult patients from the otolaryngology outpatient clinic selected at random. All possible patterns in the combination of three tests were explored, and their interpretations worked out. The findings were compared with those of pure-tone audiometry to check their sensitivity, specificity, and validity. RESULTS AND OBSERVATIONS: Fifty-eight patterns were obtained, which were classified into 10 types (5 tables) according to the type of hearing loss. The type of hearing loss was determined in all cases, and the better ear was identified in many cases. The overall sensitivity was 76.86%, the specificity was 85.48%, and the validity was 78.54%. CONCLUSIONS: The tuning fork tests of Rinne, Weber, and absolute bone conduction, when combined and interpreted, can be reliable initial diagnostic tools. They can be used to decide whether referral to a specialist or further audiometric testing is required. The patterns presented here depict the vast number of logical possibilities wherein the authenticity of these tests can be checked and the source of errors identified. PMID- 15903204 TI - Localization of T cells and subtypes in the paranasal sinus and turbinate mucosa in patients with chronic sinusitis. AB - The aim of this study was to quantitate total T lymphocytes (total CD3+ cells) and T-lymphocyte subtypes (CD4+ [T helper] and CD8+ [T suppressor] cells) in patients with chronic sinusitis who were treated with functional endoscopic sinus surgery (FESS) and to investigate the pathophysiology of persistent inflammation in chronic sinusitis. This prospective study was conducted in study and control groups. The study group consisted of 32 patients (20 male, 12 female) with chronic sinusitis who underwent FESS. The control group consisted of 8 nonsinusitis patients (5 male, 3 female) who underwent septoplasty. Specimens from the study group were excised from five regions: the uncinate process, maxillary and ethmoid sinuses, and middle and inferior turbinates. The specimens were examined with x10 magnification by light microscopy, and the slides with a severe inflammatory process were included. Punch biopsy of the control group was taken from the inferior turbinate with patients' written approval. The surgical specimens from the study and control groups were examined with an immunohistochemical staining technique with monoclonal antibodies against CD3, CD4, and CD8 surface antigens of T lymphocytes. In every specimen, the numbers of CD3+, CD4+, and CD8+ cells were calculated in 3 to 4 high magnification field on light microscopy, and the mean number of these cells in the epithelium, subepithelial layer of the lamina propria, and deep paraglandular layer of the mucosa was determined. Statistical analysis by Kruskal-Wallis analysis of variance and the Mann-Whitney U test with Bonferroni correction revealed that the CD3 epithelial layer value of the inferior turbinate (p = .030) and the CD4 deep layer value of the middle turbinate (p = .048) were significantly higher than the corresponding values of the control group. In the epithelial (p = .018) and subepithelial (p = .012) layers of the uncinate process group, in the epithelial (p = .050) and subepithelial (p = .012) layers of the ethmoid sinus group, and in the subepithelial (p = .018) and deep paraglandular (p = .012) layers of the middle turbinate group, the difference between the CD4+ and CD8+ cell counts was found to be statistically significant by the Wilcoxon signed rank test. The number of CD4+ cells was higher than the number of CD8+ cells. In conclusion, T cells play a role in the pathophysiology of chronic sinusitis. CD4+ T helper cells, in particular, are predominant at the initiation and regulation of inflammation. The uncinate process, ethmoid sinus, and middle and inferior turbinates have the main roles by T cells and subtypes in the defense system in chronic sinusitis. PMID- 15903205 TI - Internal auditory canal: pre- and postnatal growth. AB - OBJECTIVE: The purpose of this study was to describe the size and shape of the internal auditory canal (IAC) during development. STUDY DESIGN: The study was conducted in 98 dried temporal bones from 7-month-old fetuses to adults. Silicone mould casts of the IAC were used. METHODS: The length of the IAC was measured on the moulds corresponding four walls as the diameter on the fundus, middle portion, and porus. RESULTS: Although size and shape vary considerably among individuals, the data showed no significant differences between the right and left sides of the same individual. The values of the wall length and porus diameter increase as a function of age, especially for the upper wall and horizontal diameter. Both middle portion diameters suffer variations at different ages, increasing and decreasing until reaching the final shape. The diameter of the fundus, near the inner ear, remains stable. CONCLUSIONS: Our results showed that during development, the shape of the IAC adapts to cranial growth, with those of adults and fetuses having a similar shape but different dimensions, which was quite evident near the porus. PMID- 15903206 TI - Characteristics of tinnitus: investigation of over 1400 patients. AB - The data in the literature concerning tinnitus characteristics are few and contrasting. Almost all data were collected by means of questionnaires mailed to the subjects, without considering the type of tinnitus and the eventual association with other otologic symptoms. To collect, in a homogeneous way and directly from patients, personal and relevant tinnitus data, we adopted a protocol of study that allowed us to select all patients suffering from idiopathic tinnitus, to obtain a wide range of information concerning the symptoms, and to compare qualitative and quantitative tinnitus data referred by patients with those obtained through audiometry. The age at which tinnitus appears more frequently is between 40 and 50 years. No significant differences between males and females were observed. The percentage of those reporting noise exposure was low. In most cases, the duration of the tinnitus was less than 1 year and more than 5 years. Loudness matching values show a homogeneous distribution for levels between 0 and 12 dB and over 15 dB, without correspondence with the subjective judgement of tinnitus intensity. Frequencies resulting high are those between 0 and 1000 Hz and those at 8000 Hz. There is a correspondence between loudness level and masking level and between loudness level and residual inhibition. Data resulting in this study underline the importance of a global evaluation of patients suffering from tinnitus, including subjective data and tinnitus measurements. PMID- 15903207 TI - Primary tumour volume: important predictor of outcome for T3- and T4-staged nasopharyngeal carcinoma. AB - OBJECTIVE: To investigate the relationship between primary tumour volumes and treatment outcomes within T3- and T4-staged nasopharyngeal carcinoma. DESIGN: Retrospective study. SETTING: Tertiary care centre. METHODS: Forty newly diagnosed T3-staged patients and 36 newly diagnosed T4-staged patients participated in the study. MAIN OUTCOME MEASURES: Computed tomography-derived primary tumour volume was obtained from the summation of areas technique. The probabilities of achieving tumour control and patient survival were estimated using the product-limit method of Kaplan and Meier. The log rank test was used to examine significance. RESULTS: In T3-staged nasopharyngeal carcinoma, the median primary tumour volume was 29.6 mL, with a range from 8.0 to 131.8 mL. After segregating the primary tumour volume into two subgroups (< 30 mL, > 30 mL), large primary tumour volume was associated with a significantly poorer disease specific survival (p = .0001). In T4-staged cases, the median primary tumour volume was 54.07 mL, with a range from 6.7 to 223.1 mL. After segregating the primary tumour volume into two subgroups (< 60 mL, > 60 mL), larger primary tumour volume was associated with a significantly poorer disease-specific survival (p = .0022). CONCLUSION: Within the same T3- and T4-staged nasopharyngeal carcinoma, the primary tumour volume represented an important prognostic factor. To improve the treatment outcome of T3- and T4-staged nasopharyngeal carcinoma with large primary tumour volumes, more aggressive treatment is needed. PMID- 15903208 TI - Oncocytoma of the nasopharynx presenting with middle ear effusion. PMID- 15903209 TI - Segmental lower lobe collapse of the left lung associated with mediastinitis secondary to parapharyngeal abscess and quinsy. PMID- 15903210 TI - Is there a role for conservative management for symptomatic laryngopyocele? Case report and literature review. PMID- 15903211 TI - Clinical spectrum of Fraser's syndrome: case report and review of the literature. PMID- 15903212 TI - Squamous cell carcinoma in a mucosal epidermal inclusion cyst. PMID- 15903213 TI - Fourth branchial cyst presenting with right-sided vocal cord paralysis. AB - Fourth branchial anomalies are rare entities. Despite this, they must be considered when presented with a low lateral neck mass, recurrent lower neck abscesses, or recurrent suppurative thyroiditis. The truly unique presentation of this case highlights the importance of including these anomalies in the differential diagnosis for all neck lesions. PMID- 15903214 TI - Pencil pushed through a younger sibling's inferior orbital fissure: filopathic foreign body injury. PMID- 15903215 TI - Ruptured petrous carotid pseudoaneurysm complicating malignant otitis externa. PMID- 15903216 TI - [The experience of hepatic transplantation from living relative donor in Ukraine]. AB - Modern approaches to hepatic transplantation in adults and children are suggested. The data concerning the need in the hepatic transplantation performance are adduced, indications for it are established, there were also discussed the principal methods of hepatic transplantation from cadaver and from living relative donor, immunosuppression therapy issues, as well as the law and deontological problems. PMID- 15903217 TI - [Prophylaxis and treatment of intraabdominal purulent complications after performance of laparoscopic cholecystectomy using interventions with ultrasonographic control]. AB - The results of treatment of 9103 patients, to whom laparoscopic cholecystectomy was performed, were adduced. In 6714 patients intraabdominal postoperative complications were absent, ultrasonographic investigation (USI) was not performed. Dynamical USI was done in 2389 patients, permitting to estimate timely the wound course severity in the gallbladder bed, to administer an adequate therapv, in 9 of them--to perform puncture of subhepatic formations under USI control. Intraabdominal infectious complications were revealed in 65 (0.71%) of patients. PMID- 15903218 TI - [Immediate results of transabdominal rectal resection with the duplicature anastomosis formation]. AB - There were presented immediate results of surgical treatment of 425 patients with the upper ampullar and rectosygmoid portion cancer recti (CR), to whom in 1989 2002 transabdominal rectal resection, using the duplicature anastomosis, was performed. In 93.4% of observations, according to histological investigation data, adenocarcinoma was diagnosed. Concurrent diseases were revealed in 226 (53.2%) patients. The technique of transabdominal CR resection is depicted. There were noted fair immediate results in treatment of the patients: postoperative complications rate have constituted 13.2%, postoperative mortality--2.6%. In structure of postoperative complications purulent-septic one had prevailed, similarly were complications with cardiovascular and respiratory systems affections. PMID- 15903219 TI - [Computer tomography from the point of view of standards in diagnosis and treatment of an acute pancreatitis]. AB - Results of pancreatic computer tomography (CT) were studied in 162 patients, suffering an acute pancreatitis (AP). There were conducted 293 investigations in total. Basing on the obtained results analysis there were established the optimal terms of the CT conduction, the indications were-substantiated for performance of the investigation in dynamics in patients, who were or were not operated on. After the first AP attack the CT performance was indicated on the 7-10th day, after reattack--on 2-3d day of the disease. There was substantiated the necessity to conduct CT at the end of the disease first month and in the absence of pathological changes in pancreas--reinvestigation at the end of the 6th month. In revealing pathological changes of the pancreatic gland structure, it is expedient to perform dynamical observation using ultrasonographic investigation. The standard protocol for monitoring of the pancreatic gland state was elaborated, taking into account the CT role and significance. PMID- 15903220 TI - [Immunity disorders in an acute destructive pancreatitis and methods of their correction]. AB - Immune status in patients with destructive pancreatitis (DP) in long persisting disease and after performance of operative intervention was studied. Signs of secondary combined immunodeficiency were revealed, manifested by decrease of absolute quantity of all lymphocytes populations by 20-70% and concentration of immunoglobulins in blood serum by more than 40%. Conventional therapy did not influence indexes of immunogramm while using immunoglobulin for intravenous injection (IGII), besides saturation of blood by antibodies, the complement activities are increasing as well as indexes of the phagocytes functional state. Application of IGII had promoted the decrease of the patients state severity, normalization of clinico-laboratory data, causing the mortality lowering (P < 0.05), decrease of the treatment duration of survived patients (P < 0.05). PMID- 15903221 TI - [Changes in the coagulational hemostasis system in venous thromboembolism]. AB - The work is based on the results of examination of 51 patients, suffering various clinical variants of venous thromboembolism. There were adduced such indexes of the coagulational hemostasis systems, as the coagulation activation, anticoagulant and fibrinolytic. It was established, that disorders in the coagulational hemostasis system may be manifested by increase in activity of the coagulation activation system or by decrease in activity of anticoagulant or fibrinolytic system. The disorders in the coagulational hemostasis system causes the pulmonary thromboembolism occurrence. PMID- 15903222 TI - [Results of roentgenological diagnosis for revelation of patients with high surgical risk]. AB - Examination of 1142 patients with application of roentgenological method was conducted, the increase of quantity of patients with complicated thoracoabdominal diseases was established. Authentic predominance of patients with severe disease aged 40-60 years old and older was determined. Correlative connection between age of patients and the number of cases in autumn was established. Specialized help improvement with attraction of qualified professionals was recommended, taking into account the severity of the patients state, appreciating risk of surgical intervention. PMID- 15903223 TI - [The ratio of nosological forms and surgical tactics in the treatment of the thyroid gland diseases]. AB - The dynamics of morphofunctional changes of the thyroid gland tissue was analyzed in patients with the organ diseases in 1995-2003. It was established, that geochemical environmental factors constitute the cause of the nodous goiter occurrence. The environment soiling causes the thyroid gland tissue dysplasia and the cancer occurrence. The surgical tactics in the thyroid gland diseases was adduced. PMID- 15903224 TI - [Activity of the thyroid and sex hormones in patients with myasthenia after surgical intervention]. AB - In 1995-2003 yrs there were operated on 55 women for various forms of myasthenia. In 20 of them myasthenia coexisted with thymoma, in 35--have been combined with the thymus hyperplasia. In all the patients the hormonal misbalance was revealed, depending on changes in the thymus structure present. PMID- 15903225 TI - [Application of simultaneous surgeries in patients with extended pulmonary tuberculosis]. AB - The treatment of extended pulmonary tuberculosis, resistant to antibacterial preparations, constitutes extremely complex problem. The author proposes to apply simultant pulmonary resection and intrapleural thoracoplasty, the simple one or extended, for the treatment of those patients. There were studied up the results of treatment of 401 patients, simultant interventions were used in 134. Analysis of the data obtained permits to consider the operation as the curative prophylactic one. The indications for application of every variant of intervention were determined, what have had significantly increased the possibilities of treatment of extended pulmonary tuberculosis. PMID- 15903226 TI - [Evisceration: causes, prophylaxis, treatment]. PMID- 15903227 TI - [Rare complication of giant abdominal wall hernia by the small intestine perforation and the hernial sac phlegmon]. PMID- 15903228 TI - [Lipoma of ascending colon, simulating an acute appendicitis]. PMID- 15903229 TI - [An acute diverticulitis in incarcerated inguinal hernia in a child]. PMID- 15903230 TI - [Giant ovarian cystoma, simulating pseudoascitic syndrome]. PMID- 15903231 TI - [Total inferior vena cava occlusion by metastatic thrombi of Wilms tumor]. PMID- 15903232 TI - [Concomitant acute appendicitis with the right ovary apoplexy and left fallopian tube torsion]. PMID- 15903233 TI - [Report about the work of International scientific-practical conference of surgeons "Modern issues of diagnosis and surgical treatment of acute diseases of abdominal organs"]. PMID- 15903234 TI - Brief overview of BioMicroNano technologies. AB - This paper provides a brief overview of the fields of biological micro electromechanical systems (bioMEMs) and associated nanobiotechnologies, collectively denoted as BioMicroNano. Although they are developing at a very rapid pace and still redefining themselves, several stabilized areas of research and development can be identified. Six major areas are delineated, and specific examples are discussed and illustrated. Various applications of the technologies are noted, and potential market sizes are compared. PMID- 15903235 TI - Glycosylation of recombinant antibody therapeutics. AB - The adaptive immune system has the capacity to produce antibodies with a virtually infinite repertoire of specificities. Recombinant antibodies specific for human targets are established in the clinic as therapeutics and represent a major new class of drug. Therapeutic efficacy depends on the formation of complexes with target molecules and subsequent activation of downstream biologic effector mechanisms that result in elimination of the target. The activation of effector mechanisms is dependent on structural characteristics of the antibody molecule that result from posttranslational modifications, in particular, glycosylation. The production of therapeutic antibody with a consistent human glycoform profile has been and remains a considerable challenge to the biopharmaceutical industry. Recent research has shown that individual glycoforms of antibody may provide optimal efficacy for selected outcomes. Thus a further challenge will be the production of a second generation of antibody therapeutics customized for their clinical indication. PMID- 15903236 TI - Temperature reduction in cultures of hGM-CSF-expressing CHO cells: effect on productivity and product quality. AB - We have demonstrated that temperature reduction from 37 to 33 degrees C in the culture of a CHO cell line producing recombinant human granulocyte macrophage colony stimulating factor (CHO-K1-hGM-CSF) leads to a reduced growth rate, increased cell viability, improved cellular productivity, and decreased cell metabolism. In the present study, CHO-K1-hGM-CSF cells were cultured in a biphasic mode: first, a 37 degrees C growth phase for achieving a high cell number, followed by a production phase where the culture temperature was shifted to 33 degrees C. The maximum cell density was not affected after temperature reduction while cell viability remained above 80% for a further 3.7 days in the culture kept at the lower temperature, when compared to the control culture maintained at 37 degrees C. Furthermore, the total rhGM-CSF production increased 6 times in the culture shifted to 33 degrees C. Because the quality and hence the in vivo efficacy of a recombinant protein might be affected by numerous factors, we have analyzed the N- and O-glycosylation of the protein produced under both cell culture conditions using high-pH anion-exchange chromatography and complementary mass spectrometry techniques. The product quality data obtained from the purified protein preparations indicated that decreasing temperature had no significant effect on the rhGM-CSF glycosylation profiles, including the degree of terminal sialylation. Moreover, both preparations exhibited the same specific in vitro biological activity. These results revealed that the employed strategy had a positive effect on the cell specific productivity of CHO-K1-hGM CSF cells without affecting product quality, representing a novel procedure for the rhGM-CSF production process. PMID- 15903237 TI - Enhanced production of monomeric interferon-beta by CHO cells through the control of culture conditions. AB - The enhancement of recombinant protein expression of a transfected cell line is essential for the development of an efficient large-scale bioprocess. The effect of various media additives and temperature conditions were studied in an attempt to optimize protein production, stability, and protein glycosylation from a Chinese hamster ovary (CHO) cell line producing human beta-interferon (Hu-beta IFN). We observed a decrease in the ELISA response of the glycoprotein in the later stages of batch cultures, which was attributed to molecular aggregation. Cells were subjected to various concentrations of glycerol, dimethyl sulfoxide (DMSO), and sodium butyrate (NaBu) in a variety of culture systems and conditions. The addition of both NaBu and DMSO resulted in higher specific productivities but reduced growth rates that resulted in a net reduction of interferon produced. Glycerol appeared to stabilize the secreted beta-IFN, resulting in reduced aggregation, despite a decrease in cell growth rate. Glycosylation analysis of isolated beta-IFN showed a time-dependent decrease in sialylation in batch culture that was ameliorated by the presence of glycerol. Low-temperature conditions (30 degrees C) had the greatest effect on productivity with a significant increase in beta-IFN titer as well as a reduction in the degree of molecular aggregation. PMID- 15903238 TI - Production and glycosylation of recombinant beta-interferon in suspension and cytopore microcarrier cultures of CHO cells. AB - Microcarriers are suitable for high-density cultures of cells requiring surface attachment and also offer the advantage of easy media removal for product recovery. We have used the macroporous microcarriers Cytopore 1 and 2 for the growth of CHO cells producing recombinant human beta-interferon (beta-IFN) in stirred batch cultures. Although these cells may grow in suspension, in the presence of Cytopore microcarriers they become entrapped in the inner bead matrix where they can be maintained at high densities. Cell growth rates were reduced in microcarrier cultures compared to suspension cultures. However, the beta-IFN yield was up to 3-fold greater as a result of an almost 5-fold higher specific productivity. Maximum productivity was found in cultures containing 1.0 mg/mL of Cytopore 1 or 0.5 mg/mL of Cytopore 2 with a cell/bead ratio of 1029 and 822, respectively. Beta-IFN molecules aggregated in the later stages of all cultures, causing a decrease in response by ELISA. However, the degree of aggregation was significantly less in the microcarrier cultures. The N-linked glycans from beta IFN were isolated and analyzed by normal phase HPLC. There was no apparent difference in the profile of glycans obtained from each of the suspension and Cytopore culture systems. This suggests that Cytopore microcarriers may be useful in bioprocess development for enhanced recombinant glycoprotein production without affecting the glycosylation profile of the protein. PMID- 15903239 TI - Effect of production method and gene amplification on the glycosylation pattern of a secreted reporter protein in CHO cells. AB - We have investigated the independent effects of selective gene amplification (using the dhfr amplifiable selection marker) and culture operating strategy (batch vs repeated fed-batch vs semicontinuous perfusion) on the glycosylation of a recombinant reporter protein (secreted alkaline phosphatase, SEAP) produced in transfected Chinese hamster ovary (CHO) cells. HPLC analyses coupled with susceptibility to various exoglycosidases were used to determine the N glycosylation profile of SEAP samples. The dhfr amplified cell line yielded an almost 10-fold increase in specific productivity as compared to that of the unamplified cell line. The glycosylation pattern of the reporter protein produced in batch bioreactor cultures of the amplified cell line showed only slight differences as compared to the glycosylation pattern of the protein from batch bioreactor cultures of the unamplified cell line. In contrast, analysis of SEAP glycosylation structures from the protein isolated from semicontinuous perfusion cultures indicated that both relative glycan content and extent of sialylation were increased as compared to samples isolated from repeated fed-batch cultures. These results suggest that the slow growing perfusion cultures produce more completely glycosylated proteins than the faster growing repeated fed-batch cultures. PMID- 15903240 TI - Enhanced human thrombopoietin production by sodium butyrate addition to serum free suspension culture of bcl-2-overexpressing CHO cells. AB - When sodium butyrate (NaBu) was added to serum-free suspension culture of recombinant CHO (rCHO) cells for enhanced expression of human thrombopoietin (hTPO), apoptotic cell death of rCHO cells was induced in a dose-dependent manner and hTPO quality was deteriorated in regard to sialic acid and acidic isoform contents. To overcome these problems, we overexpressed Bcl-2 protein, an antiapoptotic protein, in rCHO cells producing hTPO. Compared to serum-free suspension culture of control cells without Bcl-2 overexpression (R-neo cells) and NaBu addition, a more than 10-fold increase in the maximum hTPO concentration was obtained in serum-free suspension culture of cells with Bcl-2 overexpression (R-bc12-14 cells) and 3 mM NaBu addition. Both the enhanced specific productivity endowed by NaBu and the extended culture longevity provided by the antiapoptotic effect of Bcl-2 overexpression contributed to the enhancement of maximum hTPO concentration. The problem of quality reduction of hTPO induced by NaBu was not solved by Bcl-2 overexpression, but it was not that significant. Compared to the culture in the absence of NaBu, the percentage of hTPO isoforms in pI 3-5 with high in vivo biological activity produced by R-bc12-14 cells was decreased by approximately 18% in the presence of 3 mM. As a result, a more than 6-fold increase in the production of hTPO isoforms in pI 3-5 was achieved in R-bcl2-14 cell culture with 3 mM NaBu addition. Taken together, the data obtained suggest that Bcl-2 overexpression in rCHO cells and NaBu addition in serum-free suspension culture can be an effective means to enhance the production of highly glycosylated protein such as hTPO. PMID- 15903241 TI - Substitution of glutamine by pyruvate to reduce ammonia formation and growth inhibition of mammalian cells. AB - In mammalian cell culture technology glutamine is required for biomass synthesis and as a major energy source together with glucose. Different pathways for glutamine metabolism are possible, resulting in different energy output and ammonia release. The accumulation of ammonia in the medium can limit cell growth and product formation. Therefore, numerous ideas to reduce ammonia concentration in cultivation broths have been developed. Here we present new aspects on the energy metabolism of mammalian cells. The replacement of glutamine (2 mM) by pyruvate (10 mM) supported cell growth without adaptation for at least 19 passages without reduction in growth rate of different adherent commercial cell lines (MDCK, BHK21, CHO-K1) in serum-containing and serum-free media. The changes in metabolism of MDCK cells due to pyruvate uptake instead of glutamine were investigated in detail (on the amino acid level) for an influenza vaccine production process in large-scale microcarrier culture. In addition, metabolite profiles from variations of this new medium formulation (1-10 mM pyruvate) were compared for MDCK cell growth in roller bottles. Even at very low levels of pyruvate (1 mM) MDCK cells grew to confluency without glutamine and accumulation of ammonia. Also glucose uptake was reduced, which resulted in lower lactate production. However, pyruvate and glutamine were both metabolized when present together. Amino acid profiles from the cell growth phase for pyruvate medium showed a reduced uptake of serine, cysteine, and methionine, an increased uptake of leucine and isoleucine and a higher release of glycine compared to glutamine medium. After virus infection completely different profiles were found for essential and nonessential amino acids. PMID- 15903242 TI - Effects of elevated pCO2 and osmolality on growth of CHO cells and production of antibody-fusion protein B1: a case study. AB - Partial pressure of CO2 (pCO2) and osmolality as high as 150 mmHg and 440 mOsm/kg, respectively, were observed in large-scale CHO cell culture producing an antibody-fusion protein, B1. pCO2 and osmolality, when elevated to high levels in bioreactors, can adversely affect cell culture and recombinant protein production. To understand the sole impact of pCO2 or osmolality on CHO cell growth, experiments were performed in bench-scale bioreactors allowing one variable to change while controlling the other. Elevating pCO2 from 50 to 150 mmHg under controlled osmolality (about 350 mOsm/kg) resulted in a 9% reduction in specific cell growth rate. In contrast, increasing osmolality resulted in a linear reduction in specific cell growth rate (0.008 h(-1)/100 mOsm/kg) and led to a 60% decrease at 450 mOsm/kg as compared to the control at 316 mOsm/kg. This osmolality shift from 316 to 445 mOsm/kg resulted in an increase in specific production rates of lactate and ammonia by 43% and 48%, respectively. To elucidate the effect of high osmolality and/or pCO2 on the production phase, experiments were conducted in bench-scale bioreactors to more closely reflect the pCO2 and osmolality levels observed at large scale. Increasing osmolality to 400 450 mOsm/kg did not result in an obvious change in viable cell density and product titer. However, a further increase in osmolality to 460-500 mOsm/kg led to a 5% reduction in viable cell density and a 8% decrease in cell viability as compared to the control. Final titer was not affected as a result of an apparent increase in specific production rate under this increased osmolality. Furthermore, the combined effects from high pCO2 (140-160 mmHg) and osmolality (400-450 mOsm/kg) caused a 20% drop in viable cell density, a more prominent decrease as compared to elevated osmolality alone. Results obtained here illustrate the sole effect of high pCO2 (or osmolality) on CHO cell growth and demonstrate a distinct impact of high osmolality and/or pCO2 on production phase as compared to that on growth phase. These results are useful to understand the response of the CHO cells to elevated pCO2 (and/or osmolality) at a different stage of cultivation in bioreactors and thus are valuable in guiding bioreactor optimization toward improving protein production. PMID- 15903243 TI - Insights into the central metabolism of Spodoptera frugiperda (Sf-9) and Trichoplusia ni BTI-Tn-5B1-4 (Tn-5) insect cells by radiolabeling studies. AB - The insect cell baculovirus expression vector system (BEVS) is one of the most commonly used expression systems for recombinant protein production. This system is also widely used for the production of recombinant virus and virus-like particles. Although several published reports exist on recombinant protein expression using insect cells, information dealing with their metabolism in vitro is relatively scarce. In this work we have analyzed the metabolism of glucose and glutamine, the main carbon and/or energy compounds, of the two most commonly used insect cell lines, Spodoptera frugiperda (Sf-9) and the Trichoplusia ni BTI-Tn 5B1-4 (Tn-5). Radiolabeled substrates have been used to determine the flux of glucose carbon entering the tricarboxylic acid cycle (TCA) and the pentose phosphate (PP) pathway by direct measurement of 14CO2 produced. The percentage of total glucose metabolized to CO2 via the TCA cycle was higher in the case of the Sf-9 (2.7%) compared to Tn-5 (0.6%) cells, while the percentage of glucose that is metabolized via the PP pathway was comparable at 14% and 16% for the two cell lines, respectively. For both cell lines, the remaining 83% of glucose is metabolized through other pathways generating, for example, lactate, alanine, etc. The percentage of glutamine oxidized in the TCA cycle was approximately 5 fold higher in the case of the Tn-5 (26.1%) as compared to the Sf-9 cells (4.6%). Furthermore, the changes in the metabolic fluxes of glucose and glutamine in Tn-5 PYC cells, which have been engineered to express a cytosolic pyruvate carboxylase, have been studied and compared to the unmodified cells Tn-5. As a result of this metabolic engineering, significant increase in the percentage of glucose oxidized in the TCA cycle (3.2%) as well as in the flux through the PP pathway (34%) of the Tn-5-PYC were observed. PMID- 15903244 TI - Defined protein-free NS0 myeloma cell cultures: stimulation of proliferation by conditioned medium factors. AB - A chemically defined, protein-free, and animal-component-free medium, designated RITM01, has been developed for NS0 myeloma cells. The basal medium used was a commercial serum-free and protein-free hybridoma medium, which was supplemented with phosphatidylcholine, cholesterol, beta-cyclodextrin, and ferric citrate. Increasing the amino acid concentration significantly improved cell growth. An NS0 cell line, constitutively producing a human IgG1 antibody, reached a peak cell density of 3 x 10(6) cells mL(-1) in this medium. The antibody yield was 195 mg L(-1) in batch culture, which is a 3-fold increase compared to that of a standard serum-supplemented medium, even though the cell yield was the same. The increase in antibody yield was a consequence of a longer growth phase and a slight increase in specific antibody production rate at low specific proliferation rates. Adaptation of the NS0 myeloma cell line to the protein-free conditions required about 3 weeks before viability and cell densities were stabilized. Most probably, changes in gene expression and phenotypic behavior necessary for cell survival and proliferation occurred. We hypothesize that mitogenic factors produced by the cells themselves are involved in autocrine control of proliferation. To investigate the presence of such factors, the effect of conditioned (spent) medium (CM) on cell growth and proliferation was studied. Ten-fold concentrated CM, harvested at a cell density of 2 x 10(6) cells mL(-1), had a clear positive effect on proliferation even if supplied at only 2.5% (v/v). CM was found to contain significant amounts of extracellular proteins other than the antibody. Fractionation of CM on a gel filtration column and subsequent supplementation of new NS0 cultures with the individual fractions showed that factors eluting at 20-25 kDa decreased the lag phase and increased the peak cell density as compared to control cultures. Identification of autocrine factors involved in regulation of proliferation may lead to completely new strategies for control of growth and product formation in animal cell processes. PMID- 15903245 TI - Survival factor-like activity of small peptides in hybridoma and CHO cells cultures. AB - Synthetic peptides containing three to six amino acid residues were previously shown to improve key parameters of monoclonal antibody-producing mouse hybridoma cultures. The aim of the current work was to investigate whether small peptides also exert analogous beneficial impact on a CHO-K1-derived cell line (XMK-111-10) engineered for production of the human model glycoprotein SEAP (secreted alkaline phosphatase). Similar to hybridoma cultures, growth and SEAP production profiles of CHO XMK-111-10 were modulated by peptides. Both viable cell density and SEAP production were increased by tetraalanine or by a fraction of wheat gluten hydrolysate. Whereas tetraglycine increased the peak viable cell density, the growth-suppressing tripeptide Gly-Lys-Gly significantly boosted SEAP production. All peptide-supplemented cultures showed slight improvement of culture viability during the decline phase of the batch cultures, suggesting a survival factor-like activity of the peptides. PMID- 15903246 TI - Purification and characterization of an anti-apoptotic protein isolated from Lonomia obliqua hemolymph. AB - Previously it was reported that supplementation of insect cell culture with Lonomia obliqua hemolymph could extend culture longevity (Maranga et al. Biotechnol. Prog. 2003, 19, 58-63). In this work the anti-apoptotic properties of this hemolymph in Spodoptera frugiperda (Sf-9) cell culture were investigated. The presence or absence of apoptotic cells was characterized by light microscopy, flow cytometry, and agarose gel electrophoresis. Hemolymph was fractionated by several ion exchange and gel filtration chromatographic steps for identification of the compounds responsible for this effect. Fractions exhibiting a potent anti apoptotic effect were isolated and tested in cell culture. A protein of about 51 kDa was identified, isolated, and tested for apoptosis inhibition. Addition of this purified protein to Sf-9 cultures was able to prevent apoptosis induced by nutrient depletion as well as by potent apoptosis chemical inducers such as Actinomycin D. This work confirms that the enhanced culture longevity obtained by supplementation with L. obliqua hemolymph is due to the presence of potent anti apoptotic factors. PMID- 15903247 TI - Effect of increased expression of protein disulfide isomerase and heavy chain binding protein on antibody secretion in a recombinant CHO cell line. AB - Previous work has shown that a human-antibody-producing recombinant CHO cell line did not increase its intracellular content of protein disulfide isomerase (PDI) and heavy chain binding protein (BIP) according to the increasing expression of antibody. It was also found that the intracellular assembly of light and heavy chain is a major limiting factor for overall cell specific productivity, as secretion rates improve with higher light chain expression levels and heavy chain accumulates intracellularly when too little light chain is present. As these CHO cells had a significantly lower intracellular PDI content compared to that of hybridoma cells, these results have led us to try to overcome the limitation in the posttranslational assembly in the endoplasmatic reticulum. Recombinant CHO cells were transfected with PDI or BIP alone or in combination, and the effect on intracellular light and heavy chain content and specific production rate was determined. Overexpression of BIP, both alone and in combination with PDI, reduced the specific secretion rate, whereas PDI, when overexpressed alone, caused an increase of product secretion rate. PMID- 15903248 TI - Modeling hybridoma cell metabolism using a generic genome-scale metabolic model of Mus musculus. AB - The reconstructed cellular metabolic network of Mus musculus, based on annotated genomic data, pathway databases, and currently available biochemical and physiological information, is presented. Although incomplete, it represents the first attempt to collect and characterize the metabolic network of a mammalian cell on the basis of genomic data. The reaction network is generic in nature and attempts to capture the carbon, energy, and nitrogen metabolism of the cell. The metabolic reactions were compartmentalized between the cytosol and the mitochondria, including transport reactions between the compartments and the extracellular medium. The reaction list consists of 872 internal metabolites involved in a total of 1220 reactions, whereof 473 relate to known open reading frames. Initial in silico analysis of the reconstructed model is presented. PMID- 15903249 TI - On the optimal ratio of heavy to light chain genes for efficient recombinant antibody production by CHO cells. AB - Monoclonal antibodies (Mab) are heterotetramers consisting of an equimolar ratio of heavy chain (HC) and light chain (LC) polypeptides. Accordingly, most recombinant Mab expression systems utilize an equimolar ratio of heavy chain (hc) to light chain (lc) genes encoded on either one or two plasmids. However, there is no evidence to suggest that this gene ratio is optimal for stable or transient production of recombinant Mab. In this study we have determined the optimal ratio of hc:lc genes for production of a recombinant IgG4 Mab, cB72.3, by Chinese hamster ovary (CHO) cells using both empirical and mathematical modeling approaches. Polyethyleneimine-mediated transient expression of cB72.3 at varying ratios of hc:lc genes encoded on separate plasmids yielded an optimal Mab titer at a hc:lc gene ratio of 3:2; a conclusion confirmed by separate mathematical modeling of the Mab folding and assembly process using transient expression data. On the basis of this information, we hypothesized that utilization of hc genes at low hc:lc gene ratios is more efficient. To confirm this, cB72.3 Mab was transiently produced by CHO cells at constant hc and varying lc gene dose. Under these conditions, Mab yield was increased with a concomitant increase in lc gene dose. To determine if the above findings also apply to stably transfected CHO cells producing recombinant Mab, we compared the intra- and extracellular ratios of HC and LC polypeptides for three GS-CHO cells lines transfected with a 1:1 ratio of hc:lc genes and selected for stable expression of the same recombinant Mab, cB72.3. Intra- and extracellular HC:LC polypeptide ratios ranged from 1:2 to 1:5, less than that observed on transient expression of the same Mab in parental CHO cells using the same vector. In conclusion, our data suggest that the optimal ratio of hc:lc genes used for transient and stable expression of Mab differ. In the case of the latter, we infer that optimal Mab production by stably transfected cells represents a compromise between HC abundance limiting productivity and the requirement for excess LC to render Mab folding and assembly more efficient. PMID- 15903250 TI - Recombinant antibody production by perfusion cultures of rCHO cells in a depth filter perfusion system. AB - Recombinant Chinese hamster ovary cells, producing recombinant antibody against the human platelet, were cultivated in a depth filter perfusion system (DFPS). When perfusion cultures with working volume of 1 L were operated at perfusion rates of 5/d and 6/d, volumetric antibody productivities reached values 28 and 34 times higher than that of batch suspension culture in Erlenmeyer flasks and 43 and 53 times higher than that of batch culture in a controlled stirred tank reactor, respectively. Perfusion cultures in the DFPS showed stable antibody production over the whole culture period of up to 20 days. In the DFPS, inoculated cells in suspension were entrapped in a few hours within the depth filter matrix by medium circulation and retained there until the void space of the filter matrix was saturated by the cultured cells. After cells in the depth filter matrix reached saturation, overgrown viable cells at a perfusion rate of 5/d or 6/d were continuously collected into waste medium at a density of 2-4 x 10(5) cells/mL, which resulted in stable operation at high perfusion rates, maintaining values of process parameters such as glucose/lactate concentration, pH, and dissolved oxygen concentration. Because the DFPS overcomes most drawbacks observed with conventional perfusion systems, it is preferable to be used as a key culture system to produce monoclonal antibody stably for a long culture period. PMID- 15903251 TI - A perfusion culture system using a stirred ceramic membrane reactor for hyperproduction of IgG2a monoclonal antibody by hybridoma cells. AB - A novel perfusion culture system for efficient production of IgG2a monoclonal antibody (mAb) by hybridoma cells was developed. A ceramic membrane module was constructed and used as a cell retention device installed in a conventional stirred-tank reactor during the perfusion culture. Furthermore, the significance of the control strategy of perfusion rate (volume of fresh medium/working volume of reactor/day, vvd) was investigated. With the highest increasing rate (deltaD, vvd per day, vvdd) of perfusion rate, the maximal viable cell density of 3.5 x 10(7) cells/mL was obtained within 6 days without any limitation and the cell viability was maintained above 95%. At lower deltaD's, the cell growth became limited. Under nutrient-limited condition, the specific cell growth rate (mu) was regulated by deltaD. During the nonlimited growth phase, the specific mAb production rate (qmAb) remained constant at 0.26 +/- 0.02 pg/cell x h in all runs. During the cell growth-limited phase, qmAb was regulated by deltaD within the range of 0.25-0.65 vvdd. Under optimal conditions, qmAb of 0.80 and 2.15 pg/cell x h was obtained during the growth-limited phase and stationary phase, respectively. The overall productivity and yield were 690 mg/L x day and 340 mg/L x medium, respectively. This study demonstrated that this novel perfusion culture system for suspension mammalian cells can support high cell density and efficient mAb production and that deltaD is an important control parameter to regulate and achieve high mAb production. PMID- 15903252 TI - Transient gene expression in suspension HEK-293 cells: application to large-scale protein production. AB - Recent advances in genomics, proteomics, and structural biology raised the general need for significant amounts of pure recombinant protein (r-protein). Because of the difficulty in obtaining in some cases proper protein folding in bacteria, several methods have been established to obtain large amounts of r proteins by transgene expression in mammalian cells. We have developed three nonviral DNA transfer protocols for suspension-adapted HEK-293 and CHO cells: (1) a calcium phosphate based method (Ca-Pi), (2) a calcium-mediated method called Calfection, and (3) a polyethylenimine-based method (PEI). The first two methods have already been scaled up to 14 L and 100 L for HEK-293 cells in bioreactors. The third method, entirely serum-free, has been successfully applied to both suspension-adapted CHO and HEK-293 cells. We describe here the application of this technology to the transient expression in suspension cultivated HEK-293 EBNA cells of some out of more than 20 secreted r-proteins, including antibodies, dimeric proteins, and tagged proteins of various complexity. Most of the proteins were expressed from different plasmid vectors within 5-10 days after the availability of the DNA. Transfections were successfully performed from the small scale (1 mL in 12-well microtiter plates) to the 2 L scale. The results reported made it possible to establish an optimized cell culture and transfection protocol that minimizes batch-to-batch variations in protein expression. The work presented here proves the applicability and robustness of transient transfection technology for the expression of a variety of recombinant proteins. PMID- 15903253 TI - Production of recombinant adeno-associated viral vectors using a baculovirus/insect cell suspension culture system: from shake flasks to a 20-L bioreactor. AB - Production of recombinant adeno-associated viral vectors using a baculovirus/insect cell system at various scales is presented. Shake flask studies were conducted to assess conditions to be used in bioreactors. Two insect cell lines, Trichoplusia ni (H5) and Spodoptera frugiperda (Sf9), were compared for their ability to produce rAAV-2 after infection with recombinant baculoviruses coding for the essential components of the vector. The effect of varying the ratio between individual baculoviruses and the effect of the overall multiplicity of infection (MOI), as well as the cell density at infection, were also examined. Infectious rAAV-2 particles were proportionally produced when increasing the individual MOI of BacRep virus up to 1.6. When equal amounts of each virus were used, a leveling effect occurred beyond an overall MOI of 5 and a maximum titer was obtained. Increasing the cell density at infection resulted in higher yields when infecting the cells in fresh medium; however, for the production of bioactive particles, an optimal peak cell density of approximately 1 x 10(6) cells/mL was observed without medium exchange. Infection in 3- and 20-L bioreactors was done at an overall MOI of 5 with a ratio of the three baculoviruses equal to 1:1:1. Under these conditions and infecting the cells in fresh medium, a total of approximately 2.2 x 10(12) infectious viral particles (bioactive particles) or 2.6 x 10(15) viral particles were produced in a 3-L bioreactor. Without replacing the medium at infection, similar titers were produced in 20 L. Our data demonstrates the feasibility of rAAV-2 production by BEVS at various scales in bioreactors and indicates that further optimization is required for production at high cell densities. PMID- 15903254 TI - Construction and evaluation of drug-metabolizing cell line for bioartificial liver support system. AB - Focusing on drug metabolism in liver, we constructed and evaluated a drug metabolizing bioartificial liver (BAL) support system. In a previous study, we constructed ammonia-metabolizing CHO and hepatoma-derived HepG2 cell lines by recombination of the glutamine synthetase (GS) gene. For further mimicking of liver metabolism, the human hepatoma-derived cell line HepG2 was transformed by the pBudCE-GS-CYP3A4 vector, which contains GS and drug-metabolizing CYP 3A4 genes. The constructed GS-3A4-HepG2 cell line showed 3A4 activity higher than that of human primary hepatocytes. The drug-metabolizing activity of BAL (BAL clearance) was evaluated using this cell line. The estimated clearance was higher than that of the human hepatocyte system. PMID- 15903255 TI - Cryopreservation and in vitro expansion of chondroprogenitor cells isolated from the superficial zone of articular cartilage. AB - Understanding the proliferation mechanisms of chondroprogenitor cells and their influence on cell differentiation is crucial in order to develop large-scale expansion processes for tissue engineering applications. Proliferation control mechanisms were mainly attributed to substrate limitation and cell-cell contact inhibition. The limiting substrates were found to be components of the FCS, with an optimal proliferation rate achieved in the presence of 40% FCS. In addition, the medium supply rate was found to be essential in reducing substrate limitation. In terms of FCS, 10 microL FCS cm(-2) h(-1) was the threshold feed rate required to prevent substrate limitation. Above this rate, maximum cell densities of 5.3 x 10(5) cells/cm2 were achieved, representing a 53-fold expansion. To reduce the need for high supply rates, the effect of specific growth factors was also investigated. Cell densities of 3.3 x 10(5) cells/cm2 were achieved in batch cultures using 40% FCS and 1 ng/mL TGF-beta1. Chondroprogenitor cells were expanded in this medium up to three passages without compromising their ability to differentiate and produce cartilage-like matrix in pellet cultures. In addition to substrate limitation, cell-cell contact, even at very sparse subconfluent densities, appeared capable of exerting some degree of growth inhibition. The cells exhibited deceleratory growth kinetics, characterized by a decrease of specific growth rates over time. PMID- 15903256 TI - Quorum-sensing-based toolbox for regulatable transgene and siRNA expression in mammalian cells. AB - Technologies for regulated expression of multiple transgenes in mammalian cells have gathered momentum for bioengineering, gene therapy, drug discovery, and gene function analyses. Capitalizing on recently developed mammalian transgene modalities (QuoRex) derived from Streptomyces coelicolor, we have designed a flexible and highly compatible expression vector set that enables desired transgene/siRNA control in response to the nontoxic butyrolactone SCB1. The construction-kit-like expression portfolio includes (i) multicistronic (pTRIDENT), (ii) autoregulated, (iii) bidirectional (pBiRex), (iv) oncoretro- and lentiviral transduction, and (v) RNA polymerase II-based siRNA transcription-fine tuning vectors for straightforward implementation of QuoRex-controlled (trans)gene modulation in mammalian cells. PMID- 15903257 TI - Vaccinia virus-based expression of gp120 and EGFP: survey of mammalian host cell lines. AB - Production of recombinant proteins with the vaccinia virus expression system in five mammalian cell lines (HeLa, BS-C-1, Vero, MRC-5, and 293) was investigated for protein yield and proper posttranslational modifications. Regulatory acceptance of the host cell line was taken into consideration, where Vero, MRC-5, and 293 were considered more acceptable to the regulatory authorities. Relevant process knowledge for ease of scale-up with the particular cell type was also considered. Two proteins were expressed, enhanced green fluorescent protein (EGFP) in the cytoplasm and gp120, an HIV envelope coat protein that is secreted into the culture medium. HeLa cells produced the most EGFP at 17.2 microg/well with BS-C-1 and 293 following. BS-C-1 produced the most gp120 at 28.2 microg/mL with 293 and Vero following. Therefore, of the three most appropriate cell lines (Vero, MRC-5, and 293) for production processes, the best results were obtained with 293 cells. Although MRC-5 had a very high productivity on a per cell basis, the low cell density and slow growth rate made the overall production insufficient. Because gp120 contained a significant amount of posttranslational modification, this protein, produced by the different cell lines, was further analyzed by PNGase digestion suggesting N-linked glycosylation modifications in all cell lines tested. On the basis of these results and overall process considerations, 293 cells are recommended for further production process optimization in a serum-free suspension system. PMID- 15903258 TI - Inflammatory response and apoptosis in newborn lungs after meconium aspiration. AB - An important feature of meconium-instilled newborn lungs is an inflammatory response and apoptotic cell death. It was recently demonstrated by our group and supported by several other investigators in a relatively short period of time. Apoptosis exists also in healthy lungs, but in meconium-instilled lungs its level is usually dramatically higher. Apoptosis is characterized by loss of cell function, decrease in cell size, and its morphology. Apoptosis plays an important role in normal cell life, but increased levels of apoptosis induce great damage for any tissues. Apoptosis in the lungs has been greatly overlooked for the past decade, and meconium-induced apoptosis is a relatively new event and not effectively studied at the present time. This Review summarized current knowledge regarding meconium-induced inflammation and apoptosis in newborn lungs. PMID- 15903259 TI - Influence of auxins and sucrose in monoterpenoid oxindole alkaloid production by Uncaria tomentosa cell suspension cultures. AB - Growth and alkaloid production in Uncaria tomentosa cell suspension cultures were studied in Murashige and Skoog medium supplemented with 10 microM 2,4 dichlorophenoxyacetic acid, 10 microM kinetin, and 58 mM sucrose for maintenance and with 10 microM indole-3-acetic acid, 10 microM kinetin, and 58 mM sucrose for production. A U. tomentosa pale Uth-3 cell line, cultured in the production medium, showed a reduced lag phase and a specific growth rate (mu) of 0.27 day( 1), while cells growing in the maintenance medium showed mu = 0.20 day(-1). U. tomentosa cells growing in the production medium produced monoterpenoid oxindole alkaloids (MOA) in amounts of 10.2 +/- 1.6 microg g(-1) dry weight (DW). The chemical profile of MOA produced by in vitro cell cultures was similar to that found in the plant. After 10 subcultures, maximum MOA production decreased to 2.0 +/- 0.7 microg g(-1) DW, while tryptamine alkaloids (TA) were produced with a maximum of 6.2 +/- 0.4 microg g(-1) DW. The increase of initial sucrose concentration up to 145 mM in the production medium enhanced the cell biomass by 3.2-fold (from 10.2 +/- 0.1 to 32.8 +/- 1.1 g DW L(-1)), reduced mu from 0.27 to 0.23 day(-1), and provoked a substantial accumulation of TA (23.1 +/- 4.7 microg g(-1) DW). A high sucrose concentration stimulated MOA production in the maintenance medium (2.7 +/- 0.5 microg g(-1) DW), even in the presence of 2,4 dichlorophenoxyacetic acid. PMID- 15903260 TI - Preclinical manufacture of an anti-HER2 scFv-PEG-DSPE, liposome-inserting conjugate. 1. Gram-scale production and purification. AB - A GMP-compliant process is described for producing F5cys-PEG-lipid conjugate. This material fuses with preformed, drug-loaded liposomes, to form "immunoliposomes" that bind to HER2/neu overexpressing carcinomas, stimulates drug internalization, and ideally improves the encapsulated drug's therapeutic index. The soluble, single-chain, variable region antibody fragment, designated F5cys, was produced in E. coli strain RV308 using high-density cultures. Affinity adsorption onto horizontally tumbled Streamline rProtein-A resin robustly recovered F5cys from high-pressure-disrupted, whole-cell homogenates. Two product related impurity classes were identified: F5cys with mid-sequence discontinuities and F5cys with remnants of a pelB leader peptide. Low-pressure cation exchange chromatography, conducted at elevated pH under reducing conditions, enriched target F5cys relative to these impurities and prepared a C-terminal cysteine for conjugation. Site-directed conjugation, conducted at pH 5.9 +/- 0.1 with reaction monitoring and cysteine quenching, yielded F5cys-MP-PEG(2000)-DSPE. Low-pressure size exclusion chromatography separated spontaneously formed, high-molecular weight conjugate micelles from low-molecular-weight impurities. When formulated at 1-2 mg/mL in 10 mM trisodium citrate, 10% sucrose (w/v), at pH 6.4 (HCl), the conjugate was stable when stored below -70 degrees C. Six scale-up lots were compared. The largest 40-L culture produced enough F5cys to manufacture 2,085 mg of conjugate, enough to support planned preclinical and future clinical trials. The conjugate was 93% pure, as measured by polyacrylamide gel electrophoresis. Impurities were primarily identified as product-related. Residual endotoxin, rProtein A, and genomic DNA, were at acceptable levels. This study successfully addressed a necessary step in the scale-up of immunoliposome-encapsulated therapeutics. PMID- 15903261 TI - Preclinical manufacture of anti-HER2 liposome-inserting, scFv-PEG-lipid conjugate. 2. Conjugate micelle identity, purity, stability, and potency analysis. AB - Analytical methods optimized for micellar F5cys-MP-PEG(2000)-DPSE protein lipopolymer conjugate are presented. The apparent micelle molecular weight, determined by size exclusion chromatography, ranged from 330 to 960 kDa. The F5cys antibody and conjugate melting points, determined by differential scanning calorimetry, were near 82 degrees C. Traditional methods for characterizing monodisperse protein species were inapplicable to conjugate analysis. The isoelectric point of F5cys (9.2) and the conjugate (8.9) were determined by capillary isoelectric focusing (cIEF) after addition of the zwitterionic detergent CHAPS to the buffer. Conjugate incubation with phospholipase B selectively removed DSPE lipid groups and dispersed the conjugate prior to separation by chromatographic methods. Alternatively, adding 2-propanol (29.4 vol %) and n-butanol (4.5 vol %) to buffers for salt-gradient cation exchange chromatography provided gentler, nonenzymatic dispersion, resulting in well resolved peaks. This method was used to assess stability, identify contaminants, establish lot-to-lot comparability, and determine the average chromatographic purity (93%) for conjugate lots, described previously. The F5cys amino acid content was confirmed after conjugation. The expected conjugate avidity for immobilized HER-2/neu was measured by bimolecular interaction analysis (BIAcore). Mock therapeutic assemblies were made by conjugate insertion into preformed doxorubicin-encapsulating liposomes for antibody-directed uptake of doxorubicin by HER2-overexpressing cancer cells in vitro. Together these developed assays established that the manufacturing method as described in the first part of this study consistently produced F5cys-MP-PEG(2000)-DSPE having sufficient purity, stability, and functionality for use in preclinical toxicology investigations. PMID- 15903262 TI - Kinetic modeling of brewery's spent grain autohydrolysis. AB - Isothermal autohydrolysis treatments of brewery's spent grain were used as a method for hemicellulose solubilization and xylo-oligosaccharides production. The time course of the concentrations of residual hemicelluloses (made up of xylan and arabinan) and reaction products were determined in experiments carried out at temperatures in the range from 150 to 190 degrees C using liquid-to-solid ratios of 8 and 10 g/g. To model the experimental findings concerning to brewery's spent grain autohydrolysis several kinetic models based on sequential pseudo homogeneous first-order reactions were tested. Xylan and arabinan were assumed to yield oligosaccharides, monosaccharides (xylose or arabinose), furfural, and other decomposition products in consecutive reaction steps. The models proposed provide a satisfactory interpretation of the hydrolytic conversion of xylan and arabinan. An additional model merging the two proposed models for xylan and arabinan degradation assuming that furfural was formed from both pentoses was developed and the results obtained are discussed. The dependence of the calculated kinetic coefficients on temperature was established using Arrhenius type equations. PMID- 15903263 TI - Demonstration of a strategy for product purification by high-gradient magnetic fishing: recovery of superoxide dismutase from unconditioned whey. AB - A systematic approach for the design of a bioproduct recovery process employing magnetic supports and the technique of high-gradient magnetic fishing (HGMF) is described. The approach is illustrated for the separation of superoxide dismutase (SOD), an antioxidant protein present in low concentrations (ca. 0.15-0.6 mg L( 1)) in whey. The first part of the process design consisted of ligand screening in which metal chelate supports charged with copper(II) ions were found to be the most suitable. The second stage involved systematic and sequential optimization of conditions for the following steps: product adsorption, support washing, and product elution. Next, the capacity of a novel high-gradient magnetic separator (designed for biotechnological applications) for trapping and holding magnetic supports was determined. Finally, all of the above elements were assembled to deliver a HGMF process for the isolation of SOD from crude sweet whey, which consisted of (i) binding SOD using Cu2+ -charged magnetic metal chelator particles in a batch reactor with whey; (ii) recovery of the "SOD-loaded" supports by high-gradient magnetic separation (HGMS); (iii) washing out loosely bound and entrained proteins and solids; (iv) elution of the target protein; and (v) recovery of the eluted supports from the HGMF rig. Efficient recovery of SOD was demonstrated at approximately 50-fold increased scale (cf magnetic rack studies) in three separate HGMF experiments, and in the best of these (run 3) an SOD yield of >85% and purification factor of approximately 21 were obtained. PMID- 15903264 TI - Polymer-dispersed bicontinuous cubic glycolipid nanoparticles. AB - We found that certain amphiphilic polymers such as PEO-PPO-PEO triblock copolymer (PL) can directly disperse a cubic glycolipid, 1-O-phytanyl-beta-D-xyloside (beta XP), into bicontinuous cubic nanoparticles in water medium. The use of synchrotron small-angle X-ray diffraction (SSAXD) permitted the identification of the exact structure of these dispersed particles in the colloidal state. Dynamic light scattering method was used to obtain particle size distributions. The dispersion quality and the dispersion time can be improved by co-dissolving the lipid and the polymer in a common solvent. The mean volume diameter of these dispersed colloidal particles depends on the mixing time and polymer concentration. About 5 wt % (0.18 mol %) of polymer to lipid weight was found to be sufficient to produce stable colloidal dispersions. At this polymer content and at 3 h of stirring time, the mean volume diameter of cubic colloidal particles was found to be 1.0 microm. Increase of dispersion time to 6 h reduced the colloidal particle size from 1.0 microm to 660 nm. At 3 h of mixing time, the increase of polymer content, from approximately 5 to approximately 10 wt %, reduced the particle mean diameter from 1.0 microm to 675 nm. Irrespective of these dispersion times and polymer contents, the dispersed colloidal particles exhibit predominately the Pn3m cubic phase structure, the same as that of a beta XP-water binary mixture, although a weak coexistence of Im3m cubic phase is identified in these colloidal particles. This coexistence is found to have the characteristics of a Bonnet relation, which forms convincing evidence for the infinite periodic minimal surface descriptions (IPMS). Considering the biotechnological significance, the preparation of these colloidal dispersions was carried out in a phosphate-buffered saline (PBS) system. These cubic colloidal dispersions exhibited good stability and the cubic phase structure remained intact in the PBS system. PMID- 15903265 TI - Targeted receptor trafficking affects the efficiency of retrovirus transduction. AB - We describe the development of an experimental system to test the hypothesis that the efficiency of retrovirus transduction is dependent on the pathway of virus entry into the host cell and the intracellular trafficking itinerary of the cellular receptor with which it interacts. The experimental system consists of three model target cell lines, derived from HeLa cells, that stably express one of three interleukin-2 receptor alpha chain (CD25) chimeras, TAC, TAC-CD16, and TAC-DKQTLL, which have identical extracellular domains but different intracellular trafficking itineraries, and a targeted amphotropic murine leukemia retrovirus whose envelope proteins were modified to include a binding site for TAC at their N-termini. We found that the efficiency of retrovirus transduction was affected by the distribution and trafficking itinerary of the TAC receptors. Transduction of cells that expressed TAC-DKQTLL was nearly 4-fold lower than transduction of control cells that did not express any of the TAC receptors. In contrast, transduction of cells that expressed TAC was 1.6-fold higher than transduction of control cells, whereas transduction was not significantly affected by the expression of TAC-CD16. Our results suggest that in the course of designing a targeted retrovirus it may be prudent to target only those receptors that internalize retroviruses via pathways that most efficiently support post binding steps of infection. PMID- 15903266 TI - Transduction efficiency of pantropic retroviral vectors is controlled by the envelope plasmid to vector plasmid ratio. AB - Pantropic retroviral vectors pseudotyped with vesicular stomatitis virus envelope G protein (VSV-G) are typically produced by transient transfection of the VSV-G expression plasmid because constitutive expression of VSV-G is cytotoxic. To produce pantropic vectors, the VSV-G expression plasmid and the vector plasmid are cotransfected into a packaging cell line, such as 293-gag-pol. Typically, the ratio of VSV-G plasmid to the vector plasmid ranges from 0.33 to 1.0. However, it is not clear that this range is optimal for vector production. In this study we have systematically examined the effect of the ratio of VSV-G plasmid (pVSV-G) to vector plasmid on vector production. For this, 293-gag-pol stable packaging cells were cotransfected with pVSV-G and an enhanced green fluorescent protein- (EGFP-) expressing retroviral vector plasmid (pLTR-EGFP) by use of lipofectamine. Vector was collected following transfection and used to transduce three target cell lines, namely, 3T3 fibroblasts, telomerase-immortalized human diploid fibroblasts (HDF), and the human hepatoma cell line HuH7. Transduction efficiency was evaluated for vectors produced at different pVSV-G:pLTR-EGFP ratios such that the total amount of plasmid transfected into 293-gag-pol cells was kept constant. Our results indicate that transduction efficiency is greatest when the pVSV-G:pLTR EGFP ratio is substantially below 1.0. For 3T3 and HDF cells, the maximum transduction efficiency was obtained when a ratio of pVSV-G:pLTR-EGFP ranging from 0.053 to 0.2 was used for transfection. The relative magnitude of this effect was greater for lower transduction efficiencies in control cultures. For HuH7 cells, the beneficial effects were smaller than those observed when HDF or 3T3 cells were used. The difference in transduction efficiency for vector produced under various pVSV-G:pLTR-EGFP ratios was not due to differences in the proliferation of packaging cells or target cells. Further characterization showed that the amount of vector RNA relative to p30gag decreased as the ratio of pVSV G:pLTR-EGFP increased. These results indicate that transduction efficiency increases with increasing levels of vector RNA as long as a minimally sufficient level of pantropic envelope protein is expressed. PMID- 15903267 TI - Use of operating windows in the assessment of integrated robotic systems for the measurement of bioprocess kinetics. AB - This study examines the utility of an automated liquid handling robot integrated with a microwell plate reader to enable the rapid acquisition of bioprocess kinetic data. The relationship between the key parameters for liquid handling accuracy and precision and the sample detection period has been characterized for typical low-viscosity (<2.0 mPa x s) aqueous and organic phases and for a high viscosity aqueous phase (60 mPa x s), all exhibiting Newtonian rheology. The use of a simple graphical method enables the suitability of a given automation platform to be assessed once the user has determined the minimum sample detection period and the minimum accurate and precise dispense volume. This provides for a reduction in the duration of any experiment by maximizing well usage within each microwell plate. The suitability of employing an integrated automation platform to gather kinetic data for systems typical of those encountered in bioprocessing is analyzed via a series of case studies. Application to alkaline cell lysis, where disruption is complete within 120 s, showed that the range of available dispense volumes and the number of wells that can be utilized is limited. In contrast, analysis of a system exhibiting slow process kinetics, the fermentation of Escherichia coli TOP10 pQR239 in microwell plates, demonstrated that, for a typical sample detection period of 30 min, the only restrictions on the degree of well utilization are the liquid handling accuracy and precision and the volume capacity of the liquid handling robot. Finally, liquid-liquid extraction, an example of a kinetically independent operation, was also examined. In this case, only a single equilibrium measurement is required, which means that the only restrictions to the utilization of the integrated devices are the liquid handling accuracy and precision. Integrated automation platforms represent a powerful process development tool over traditional experimental methods used for bioprocess development. Smaller volumes of reagent and sample can be used to achieve greater throughput, while high levels of reproducibility and sensitivity are maintained. PMID- 15903268 TI - Elastic properties of the cell wall of Aspergillus nidulans studied with atomic force microscopy. AB - Currently, little is known about the mechanical properties of filamentous fungal hyphae. To study this topic, atomic force microscopy (AFM) was used to measure cell wall mechanical properties of the model fungus Aspergillus nidulans. Wild type and a mutant strain (deltacsmA), lacking one of the chitin synthase genes, were grown in shake flasks. Hyphae were immobilized on polylysine-coated coverslips and AFM force--displacement curves were collected. When grown in complete medium, wild-type hyphae had a cell wall spring constant of 0.29 +/- 0.02 N/m. When wild-type and mutant hyphae were grown in the same medium with added KCl (0.6 M), hyphae were significantly less rigid with spring constants of 0.17 +/- 0.01 and 0.18 +/- 0.02 N/m, respectively. Electron microscopy was used to measure the cell wall thickness and hyphal radius. By use of finite element analysis (FEMLAB v 3.0, Burlington, MA) to simulate AFM indentation, the elastic modulus of wild-type hyphae grown in complete medium was determined to be 110 +/- 10 MPa. This decreased to 64 +/- 4 MPa for hyphae grown in 0.6 M KCl, implying growth medium osmotic conditions have significant effects on cell wall elasticity. Mutant hyphae grown in KCl-supplemented medium were found to have an elastic modulus of 67 +/- 6 MPa. These values are comparable with other microbial systems (e.g., yeast and bacteria). It was also found that under these growth conditions axial variation in elastic modulus along fungal hyphae was small. To determine the relationship between composition and mechanical properties, cell wall composition was measured by anion-exchange liquid chromatography and pulsed electrochemical detection. Results show similar composition between wild-type and mutant strains. Together, these data imply differences in mechanical properties may be dependent on varying molecular structure of hyphal cell walls as opposed to wall composition. PMID- 15903269 TI - Affinity ligand selection from a library of small molecules: assay development, screening, and application. AB - A facile and cost-effective process for screening synthetic libraries for an affinity ligand is described. A high throughput 96-well plate filtration method was designed to screen both discrete compounds and mixtures of compounds attached to a solid support. Human serum albumin (HSA) was used as a target protein to demonstrate the proof of concept. Detection and quantitation by fluorescence was accomplished with the use of fluorescamine to conjugate the protein in the filtrate. It is found that mixtures demonstrating low average binding reflect an overall lower hit rate of the components, whereas deconvolution of mixtures with high protein binding consistently provides a high hit rate. This differs from many of the previous experiences screening solid-phase mixtures in which high false positive rates are noted to occur. A total of 100K compounds were tested: 25K as discrete samples and 75K as mixtures. An overall hit rate of 8% was observed. Secondary screening of compounds measured specificity, recovery, and dynamic binding capacity. The effectiveness of the method is illustrated using an affinity column made with a representative lead compound. A similar purity was achieved in a single-step purification of HSA from serum as compared to that obtained by two steps of ion-exchange chromatography. The process for primary screening of a large number of compounds is simple, inexpensive, and applicable to any soluble target protein of known or unknown function from crude mixtures and may have additional utility as a generic chemical affinity tool for the functional characterization of novel proteins emerging from proteomics work. PMID- 15903270 TI - Pseudomonas fluorescens 134 as a biological control agent (BCA) model in cell immobilization technology. AB - Antifungal activity against Rhizoctonia solani was achieved in vivo through the application of Pseudomonas fluorescens strain 134 encapsulated in sodium alginate beads of different sizes (0.5, 1, and 2 mm). The activity was compared to that obtainable with chemical treatments and bead-derived liquid formulations. The latter was obtained by dissolving alginate beads of 1 and 0.5 mm in 1% Na-citrate solution before application, without any significant (P < 0.05) reduction of bacterial numbers during the dissolution process. The dry bead formulations were applied next to the seeds in plant inoculation experiments, resulting in a reduction of disease symptoms, which were markedly reduced when the liquid formulation was applied. Moreover, the rate of disease symptoms related to liquid formulations from both 1 and 0.5 mm beads was comparable (near to 10%) to that of chemical treatment. Pseudomonas fluorescens strain 134 delivered as both dry and liquid formulations was able to colonize cotton root at a population density of about 10(8) CFU/g fresh root, 15 days after sowing. PMID- 15903271 TI - Loofa sponge as a scaffold for culture of rat hepatocytes. AB - The dried fruit from Luffa cylindrica (loofa sponge, LS), which represents a new chitinous source material, was used as a 3-D scaffold for the culture of rat hepatocytes. With the macroporous structure and large pore size (ca. 800 microm) of LS, cell loading to the scaffold should be carried out by dynamic seeding with continuous shaking throughout the seeding period. Hepatocytes attach well to the surface of loofa fibers after seeding and maintain their round shapes. The initial ammonia removal and urea-N synthesis rates of hepatocytes immobilized within LS slightly decreased with increasing cell densities, but their metabolic activities were comparable to or better than those in monolayer culture on tissue culture polystyrene control surfaces. Both urea-N synthesis and albumin secretion rates could be maintained up to 7 days for cells immobilized within LS and spheroid-like cell aggregates could be found after the second day. PMID- 15903272 TI - It's leadership--stupid! PMID- 15903273 TI - Kitchen table wisdom: a Freirian approach to medication adherence. AB - Most interventions to promote medication adherence are based on psychological theories of individual behavior. In contrast, this article describes the theory and practice of a socially based adherence intervention that is guided by the educational principles of Paolo Freire. This approach asserts that adherence is influenced by the patient's social context and attempts to improve adherence through identifying social constraints on adherence behavior. The program builds on the traditions of patient education through home nursing visits. Using a dialectic process of dialogue and problem solving and working with a team that includes a nurse and a peer-educator, patients are encouraged to act to change their social environment to support their desire to achieve high levels of medication adherence. This strategy does not replace, but rather supplements, traditional methods of understanding individual patient behavior and allows the patient and the nurse to consider potential solutions to adherence challenges in the larger social context. PMID- 15903274 TI - Suffering, shame, and silence: the stigma of HIV/AIDS. AB - HIV/AIDS, especially in the context of poverty, results in considerable suffering. The issues surrounding prevention, transmission, and mitigation are complex, but one very important concept sustaining the epidemic is stigma. This article examines the meaning of stigma in the literature and through the experience of people living in a high-prevalence area. An ethnographic study in rural Zimbabwe, where approximately one third of adults are infected, revealed how stigma, suffering, shame, and silence are mutually supporting concepts that challenge health promotion efforts. For a reduction in HIV/AIDS morbidity and mortality rates, there is a need to understand and act on contextual issues such as stigma with increased political and social commitment at local, national, and international levels. Nurses and other health care professionals need to be involved to ensure public policy and local interventions are aimed at enhancing supportive environments and reducing suffering. PMID- 15903275 TI - Factors influencing HIV-risk behaviors among HIV-positive urban African Americans. AB - Urban African Americans are disproportionately affected by HIV, the virus associated with AIDS. Although incidence and mortality appear to be decreasing in some populations, they continue to remain steady among inner-city African Americans. A major concern is the number of HIV-positive individuals who continue to practice high-risk behaviors. Understanding factors that increase risks is essential for the development and implementation of effective prevention initiatives. Following a constructionist epistemology, this study used ethnography to explore social and cultural factors that influence high-risk behaviors among inner-city HIV-positive African Americans. Leininger's culture care diversity and universality theory guided the study. Individual qualitative interviews were conducted with HIV-positive African Americans in the community to explore social and cultural factors that increase HIV-risky behaviors. For this study, family/kinship, economic, and education factors played a significant role in risky behaviors. Reducing HIV disparity among African Americans is dependent on designing appropriate interventions that enhance protective factors. Clinicians providing care to HIV-positive individuals can play a key role in reducing transmission by recognizing and incorporating these factors when designing effective prevention interventions. PMID- 15903276 TI - Community-based HIV education and prevention workers respond to a changing environment. AB - The purpose of this study was to understand the culture, values, skills and activities of staff involved in education and prevention activities in community based AIDS Service Organizations (ASOs) in Ontario, Canada, and to understand the role of evaluation research in their prevention programming. In this qualitative study, 33 staff members from 11 ASOs participated in semi-structured interviews that were analyzed using the grounded theory approach. ASO staff experience tension between a historical grassroots organizational culture characterized by responsiveness and relevance and a more recent culture of professionalization. Target populations have changed from being primarily gay men to an almost unlimited variety of communities. Program emphasis has shifted from education and knowledge dissemination to a broadly based mandate of health promotion, community development, and harm reduction. Integration of evidence of effectiveness, social behavioral theory, or systematic evaluation is uncommon. Understanding these points of tension is important for the nursing profession when it is engaged with ASOs in programming or evaluation research. PMID- 15903277 TI - Health care providers' and patients' perspectives on care in HIV ambulatory clinics across Ontario. AB - This descriptive study represents one component of a larger project that examined the perceptions of current and best-care practices in HIV ambulatory clinics across Ontario by health care providers and patients living with HIV/AIDS. Focus groups were held with providers and patients at eight clinics. Results showed that providers' and patients' perceptions were similar. Participants were able to describe current care practices and identify two elements of best care: patient focused care and access to care. However, both health care providers and patients acknowledged that financial constraints, appointment scheduling, and distance to clinics were some of the barriers to achieving best care. Case management and shared-care schemes are two strategies that are proposed to meet the challenge of providing collaborative integrated care that is accessible and equal to all, while still maintaining positive patient outcomes. PMID- 15903278 TI - Development of nursing guidelines for administration of enfuvirtide. PMID- 15903279 TI - Assessment of oral bisphosphonate use in elderly patients with varying degrees of kidney function. AB - BACKGROUND: Oral bisphosphonates are prescribed for millions of men and women in the United States who have osteopenia or osteoporosis. However, there are limited data on the use of oral bisphosphonates in patients with kidney impairment. OBJECTIVE: The purpose of this study was to evaluate changes in calculated creatinine clearance (CrCl) and reported adverse events in elderly patients with varying degrees of kidney function who were taking oral bisphosphonate therapy. METHODS: The present study was a retrospective review of the medical records of elderly patients (age range, 65-89 years) who were prescribed oral bisphosphonates for a diagnosis of osteoporosis or osteopenia at the University of Colorado Hospital Seniors Clinic during the period January 1999 through March 2003. Data on age, sex, body weight, height, blood urea nitrogen and serum creatinine concentrations, and adverse-reaction history were collected. Clinic notes were reviewed to determine whether patients had taken an oral bisphosphonate previously or whether bisphosphonate therapy was initiated during the study period. The Cockcroft-Gault equation was used to calculate CrCl. Rates of drug-related adverse events were compared in patients with and without severe kidney impairment. For the purposes of this study, severe kidney impairment was defined as a CrCl <35 mL/min. RESULTS: Data from 181 patients (88.4% female, 11.6% male; mean age, 79.2 years) who received oral bisphosphonates were included in the analysis. One hundred fourteen (63.0%) patients began bisphosphonate therapy during the study period. The mean duration of bisphosphonate therapy was 22.3 months. Thirty-one (17.1%) patients taking bisphosphonates had severe kidney impairment. There was no change in calculated CrCl after the initiation of oral bisphosphonate therapy in patients who started therapy during the study period. Adverse drug events, 19 of them related to the gastrointestinal system, occurred in 24 (13.3%) patients. Overall, 19.4% (6/31) of patients with severe kidney impairment and 12.0% (18/150) of those without severe kidney impairment experienced adverse events; the difference between groups was not statistically significant. No reported adverse drug event required hospitalization or emergency treatment. CONCLUSIONS: When prescribed for elderly patients with varying degrees of kidney function, oral bisphosphonate therapy was well tolerated and was not associated with a decline in calculated CrCl. PMID- 15903280 TI - A pharmacological surveillance study of the tolerability of policosanol in the elderly population. AB - BACKGROUND: Policosanol is a drug derived from sugar cane wax that has cholesterol-lowering and antiplatelet properties. Randomized, controlled studies are the gold standard for demonstrating drug efficacy, safety, and tolerability, but postmarketing surveillance studies are encouraged for corroborating drug effects. A valid proof of the safety of a drug is a well-documented, good tolerability profile in older individuals, since this population is more prone to drug-related adverse events (AEs). OBJECTIVE: This study investigated the tolerability of policosanol in the elderly population by monitoring the incidence and nature of AEs occurring in older Cuban patients treated with policosanol in routine clinical practice. METHODS: All patients aged > or =60 years treated with policosanol at 7 major medical centers from January 2000 to May 2003 were included. Policosanol (5, 10, or 20 mg/d) was prescribed to patients eligible to receive cholesterol-lowering and/or antiplatelet drugs, with the dosage recommended according to their individual atherosclerotic risk. Patients had follow-up visits approximately every 6 months. Data on AEs and other relevant information, including changes in policosanol treatment, concomitant medications, and discontinuations, were recorded on individual case-report forms. RESULTS: This study included 2252 patients (1306 women, 946 men): 647 (28.7%), 244 (10.8%), and 173 (7.7%) patients had coronary, cerebrovascular, and peripheral artery disease, respectively. A total of 1485 patients had hypercholesterolemia (65.9%), 1322 (58.7%) had hypertension, and 323 (14.3%) had diabetes mellitus. Of the enrolled patients, 1123 (49.9%), 644 (28.6%), and 485 (21.5%) received policosanol 5, 10, and 20 mg/d, respectively. Treatment duration varied: 2169 (96.3%), 1861 (82.6%), 1116 (49.6%), and 412 (18.3%) patients were treated for 6, 12, 24, and 36 months, respectively. Thirty-one patients (1.4%) experienced serious AEs, 18 of them fatal. Death was most often due to vascular events: myocardial infarction (4 patients), sudden cardiac arrest (1), ventricular arrhythmia (2), ischemic stroke (1), lung thromboembolism (1), cancer (5), pneumonia (1), peritonitis (1), lung edema (1), and dehydration (1). Another 13 patients (0.6%) were hospitalized, and 61 (2.7%) reported moderate or mild AEs. Overall, 21 patients (0.9%) discontinued prematurely from the study, 18 of them due to a fatal serious AE. CONCLUSIONS: Long-term tolerability of policosanol in elderly patients at high vascular risk was very good, as assessed under conditions of routine clinical practice. These results are consistent with those obtained in randomized, double-blind clinical studies of older patients treated with policosanol. PMID- 15903281 TI - The influence of celecoxib on muscle fatigue resistance and mobility in elderly patients with inflammation. AB - BACKGROUND: Acute inflammation has a negative effect on the muscular system in elderly patients, compromising the outcome of the underlying disease. OBJECTIVE: The aim of this study was to evaluate the effect of cyclooxygenase-2 (COX-2) inhibition on muscle performance and mobility in hospitalized elderly patients with acute inflammation of infectious origin. METHODS: In this single-blind, controlled trial, consecutively hospitalized elderly patients (age > or = 70 years) with inflammation (C-reactive protein [CRP] levels > or =10 mg/L) due to acute infection were randomly assigned to receive 2 weeks of treatment with the COX-2-selective inhibitor celecoxib, acetaminophen, or no supplementary medication (control). The following variables were assessed at baseline and at 1 and 2 weeks' follow-up: muscle fatigue resistance (primary outcome measure); grip strength and mobility (secondary outcome measures); and levels of the acute-phase markers CRP, interleukin (IL)-1beta, IL-6, IL-10, tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta (TGF-beta) as explanatory variables. RESULTS: Forty-three consecutively hospitalized elderly patients (31 women, 12 men; mean [SD] age, 84 [6] years) were enrolled. Fourteen patients received celecoxib, 14 received acetaminophen, and 15 received no supplementary medication. The change in fatigue resistance was significantly different between groups (P = 0.021, Kruskal-Wallis chi-square test), with significantly greater improvement in patients receiving celecoxib compared with the acetaminophen and control groups (63% increase from baseline; P < 0.05). There were no significant between-group differences in changes in grip strength, mobility, IL-1beta, IL-6, TNF-alpha, or TGF-beta. The changes in levels of IL-10 differed significantly between groups (P = 0.020, Kruskal-Wallis chi-square test), with greater improvement in the celecoxib group compared with the acetaminophen group (P = 0.032). CONCLUSION: The results of this study suggest that COX-2-selective inhibition has a beneficial effect on muscle fatigue resistance in hospitalized elderly patients with acute inflammation of infectious origin. However, until further trials are conducted, the use of COX-2-selective inhibitors for this indication is not recommended. PMID- 15903282 TI - Comparison of the association between disease burden and inappropriate medication use across three cohorts of older adults. AB - BACKGROUND: Use of potentially inappropriate medications is common in nursing facilities (NFs), in which frail older adults are particularly vulnerable to adverse drug effects. The community-dwelling elderly are generally healthier and have lower overall rates of medication use, but their prescribed medications are not subjected to the same degree of regulatory scrutiny as those of residents in NFs. Frail elderly (FE) adults who are nursing home eligible but are receiving home- and community-based services (HCBS) constitute a distinct group sharing a high disease burden and high levels of medication use with the NF population. OBJECTIVE: The goal of this study was to examine the relationship between disease burden and inappropriate medication use in these 3 cohorts, with adjustment for demographic and clinical differences. METHODS: We performed retrospective analyses of Medicaid claims data from May 2000 through April 2001 to identify 3 cohorts of Kansas Medicaid beneficiaries: community-dwelling older adults (the ambulatory cohort); persons receiving HCBS through the Kansas Frail Elderly Program (the FE cohort); and elderly NF residents (the NF cohort). Demographic, clinical, and medication data were extracted from the Medicaid claims data. Unconditionally inappropriate medications were identified using the 1997 Beers criteria. The Cumulative Illness Rating Scale for Geriatrics was used to calculate the disease burden sum, classified as 0 or 1, 2 or 3, 4 or 5, or > or =6 disease categories. Odds ratios for inappropriate medication use at each level of disease burden in each cohort were derived using multivariable models adjusted for demographic and clinical factors, including overall level of medication use. RESULTS: The final sample included 3185 persons in the 3 cohorts (1163 ambulatory, 858 FE, 1164 NF). Inappropriate medication use was determined to have occurred in 21%, 48%, and 38% of the respective cohorts and was highest in FE cohort members with the greatest disease burden (61%). For the ambulatory and FE cohorts, inappropriate medication use rose as the disease burden increased. The same was not observed in the NF cohort, in whom rates of inappropriate medication use showed little variation regardless of disease burden. CONCLUSIONS: The relationship between disease burden and inappropriate medication use varied by setting. Those members of the FE cohort with the highest disease burden had the greatest risk for inappropriate medication use. PMID- 15903283 TI - A pharmacy management intervention for optimizing drug therapy for nursing home patients. AB - BACKGROUND: A drug therapy management service was designed to reduce polypharmacy among Medicaid recipients. This service selectively focused on patients who were high users of prescription drugs and had potential drug therapy problems (PDTPs). OBJECTIVES: This article reports the results of the first phase of the North Carolina Polypharmacy Initiative. The goals of this study were to determine: (1) the frequency with which recommendations were made by pharmacists in response to targeted profile alerts aimed at high-risk patients, (2) the frequency and type of drug therapy changes, and (3) the impact on drug-related quality and costs. METHODS: A before-after design was used. Nursing home patient profiles with PDTP alerts for specific drugs and drug categories were provided to consultant pharmacists. Targeted patients had received 218 prescription fills within 90 days. Pharmacists were compensated for performing and documenting targeted drug regimen reviews. Interventions of pharmacists and results after physician consultation are described, and cost impacts of changes in drug therapy are reported. Monetary results are shown in year-2002 U.S. dollars. RESULTS: Prescription profiles were generated from Medicaid claims data and sent to consultant pharmacists for 9208 patients in 253 nursing homes. Pharmacists returned 7548 (82%) of all profiles sent to them. After excluding 1204 patients (13%) who were discharged or deceased, 6344 patients (69%) remained for analysis. At baseline, patients used a mean (SD) of 9.52 prescriptions per month, costing the North Carolina Medicaid program a mean (SD) of 502.96 dollars (309.70). A mean of 1.58 recommendations were offered to prescribers. After physician consultation, > or =1 recommendation was implemented for 72% of patients with a change recommendation, 68% of whom experienced a switch to a lower-cost drug. Drug cost savings were a mean of 30.33 dollars/patient per month. Cost savings from 1 month alone covered the compensation paid to pharmacists for consultation efforts. CONCLUSIONS: This supplemental program of medication reviews for targeted nursing home patients resulted in a reduction of polypharmacy and was beneficial based solely on drug cost savings. PMID- 15903284 TI - Does the addition of a pharmacist transition coordinator improve evidence-based medication management and health outcomes in older adults moving from the hospital to a long-term care facility? Results of a randomized, controlled trial. AB - BACKGROUND: Poorly executed transfers of older patients from hospitals to long term care facilities carry the risk of fragmentation of care, poor clinical outcomes, inappropriate use of emergency department services, and hospital readmission. OBJECTIVE: This study was conducted to assess the impact of adding a pharmacist transition coordinator on evidence-based medication management and health outcomes in older adults undergoing first-time transfer from a hospital to a long-term care facility. METHODS: This randomized, single-blind, controlled trial enrolled hospitalized older adults awaiting transfer to a long-term residential care facility for the first time. Patients were randomized either to receive the services of the pharmacist transition coordinator (intervention group) or to undergo the usual hospital discharge process (control group). The intervention included medication-management transfer summaries from hospitals, timely coordinated medication reviews by accredited community pharmacists, and case conferences with physicians and pharmacists. The primary outcome was the quality of prescribing, measured using the Medication Appropriateness Index (MAI). Secondary outcomes were emergency department visits, hospital readmissions, adverse drug events, falls, worsening mobility, worsening behaviors, increased confusion, and worsening pain. RESULTS: One hundred ten older adults (67 women, 43 men; mean [SD] age, 82.7 [6.4] years) were recruited from 3 metropolitan hospitals and assigned to 85 metropolitan long-term care facilities. Fifty-six patients were randomized to the intervention group and 54 to the control group; 44 patients in each group were evaluable at 8-week follow up. There were no significant differences in baseline characteristics between treatment groups, with the exception of the number of medications discontinued during hospitalization: a mean of 1.1 more drugs was discontinued in the control group compared with the intervention group (P = 0.011). The majority of patients (35 [62.5%] in the intervention group, 41 [76.0%] in the control group) changed physicians as part of the transition to a long-term care facility. At 8-week follow-up, there was no change in MAI from baseline in the intervention group, whereas it had worsened in the control group (mean [95% CI], 2.5 [1.4-3.7] vs 6.5 [3.9-9.1], respectively; P = 0.007). Patients who received the intervention and were alive at follow-up exhibited a significant protective effect of the intervention against worsening pain (relative risk ratio [95% CI], 0.55 [0.32 0.94]; P = 0.023) and hospital usage (i.e., the combination of emergency department visits and hospital readmissions) (0.38 [0.15-0.99]; P = 0.035), but did not differ from control patients in terms of adverse drug events (1.05 [0.66 1.68]), falls (1.19 [0.71-1.99]), worsening mobility (0.39 [0.13-1.15]), worsening behaviors (0.52 [0.25-1.10]), or increased confusion (0.59 [0.28 1.22]). When data for patients who had died were included, the intervention had no effect on hospital usage in all patients (0.58 [0.28-1.21]). CONCLUSIONS: Older people transferring from hospital to a long-term care facility are vulnerable to fragmentation of care and adverse events. In this study, use of a pharmacist transition coordinator improved aspects of inappropriate use of medicines across health sectors. PMID- 15903285 TI - Anticoagulation influences long-term outcome in patients with nonvalvular atrial fibrillation and severe ischemic stroke. AB - BACKGROUND: Limited data exist regarding long-term prognosis in patients with nonvalvular atrial fibrillation (AF) who have survived a severe, disabling stroke. OBJECTIVE: The aim of this study was to assess long-term prognosis and its determinants in a prospective case series of stroke survivors with AF and moderate to severe handicap. METHODS: From a consecutive series of AF patients with first-ever ischemic stroke, we evaluated prospectively those with moderate to severe disability (grade 4-5 on the modified Rankin Scale) who were treated during a 5-year follow-up period with either warfarin or aspirin. Death and recurrent vascular events were documented. RESULTS: Out of a pool of 438 AF patients, 191 were prospectively assessed. During a mean follow-up of 50.4 months, the cumulative 5-year mortality was 76.7% (95% CI, 69.0-84.3) and the 5 year recurrence rate was 33.7% (95% CI, 23.3-44.1). Cox regression analysis revealed that increasing age, increasing handicap, and aspirin versus warfarin were independent predictors of mortality. Prior transient ischemic attack and aspirin versus warfarin were predictors of vascular recurrence. Anticoagulation was associated with a decreased risk of death (hazard ratio [HR], 0.44; 95% CI, 0.27-0.70; P < 0.001) and recurrent thromboembolism (HR, 0.36; 95% CI, 0.17-0.77; P < 0.01). CONCLUSION: Our results suggest that chronic anticoagulation therapy may be effective in lengthening survival and preventing recurrent thromboembolism in AF patients who have suffered a severely disabling ischemic stroke. PMID- 15903286 TI - Pharmacokinetics in older persons. AB - BACKGROUND: Physiologic changes and disease-related alterations in organ function occur with aging. These changes can affect drug pharmacokinetics in older persons. OBJECTIVE: This article reviews age-related changes in pharmacokinetics and their clinical relevance. METHODS: A PubMed search was conducted using the terms elderly and pharmacokinetics. Other reviews were also included for literature searching. The review includes literature in particular from 1990 through April 2004. Some articles from before 1990 were included to help illustrate principles of age-related pharmacokinetics. RESULTS: There are minor changes in drug absorption with aging. The effect of aging on small-bowel transporter systems is not yet fully established. Bioavailability of highly extracted drugs often is increased with age. Transdermal absorption may be delayed, especially in the case of water-soluble compounds. Fat-soluble drugs may distribute more widely and water-soluble drugs less extensively in older persons. Hepatic drug metabolism shows wide interindividual variation, and in many cases, there is an age-related decline in elimination of metabolized drugs, particularly those eliminated by the cytochrome enzyme system. Any decrement in cytochrome enzyme metabolism appears nonselective. Synthetic conjugation metabolism is less affected by age. Pseudocapillarization of the sinusoidal endothelium in the liver, restricting oxygen diffusion, and the decline in liver size and liver blood flow may influence age-related changes in rate of hepatic metabolism. Frailty, physiological stress, and illness are important predictors of drug metabolism in older individuals. Inhibition of drug metabolism is not altered with aging, but induction is reduced in a minority of studies. Renal drug elimination typically declines with age, commensurate with the fall in creatinine clearance. Renal tubular organic acid transport may decline with age, while the function of the organic base transporter is preserved but may be less responsive to stimulation. CONCLUSION: Changes in pharmacokinetics occur due to age-related physiologic perturbations. These changes contribute to altered dose requirements in older persons, particularly in the case of drugs eliminated by the kidney. Interindividual variation, disease, frailty, and stress may overshadow age related changes. PMID- 15903287 TI - Efficacy and tolerability of memantine in the treatment of dementia. AB - BACKGROUND: Until recently, acetylcholinesterase inhibitors were the only approved agents for the treatment of Alzheimer's disease (AD). These medications have also been used in the treatment of vascular dementia (VD). Memantine, the first N-methyl-D-aspartate (NMDA)-receptor antagonist to be well tolerated, has been approved for the treatment of moderate to severe AD. OBJECTIVE: The aim of this study was to review the current literature on the efficacy and tolerability of memantine in the treatment of AD and VD. METHODS: A MEDLINE search of the English-language literature from January 1970 to March 2004 was conducted to identify randomized, double-blind, placebo-controlled, parallel-group trials in which memantine was administered to patients with VD or AD. The search terms were memantine, NMDA inhibitor, and NMDA antagonist. RESULTS: Excessive glutamate, the brain's major excitatory neurotransmitter, can cause excitotoxicity by allowing too much calcium to enter neuronal cells. Moderate-affinity NMDA-receptor antagonists such as memantine block pathologic activity of glutamate while allowing physiologic activity. Use of memantine has been associated with significant improvements in measures of cognition, function, and behavior in both VD and AD. Adverse events associated with memantine have been comparable to those with placebo, with the exception of an increased incidence of dizziness, constipation, cataracts, nausea, dyspnea, confusion, headache, and urinary incontinence. CONCLUSIONS: Memantine seems to be promising and well tolerated in the treatment of moderate to severe VD or AD, either as monotherapy or in combination with donepezil. It appears to be particularly effective in improving cognitive, functional, and global outcomes in moderate to severe AD and in improving cognitive end points in mild to moderate VD. More research is needed on important clinical questions, including whether memantine can prolong patients' ability to provide self-care and delay institutional placement. PMID- 15903288 TI - Cardiovascular risk and diabetes mellitus management: diagnosis and treatment in special patient populations. Introduction. PMID- 15903289 TI - Epidemiology of diabetes mellitus and associated cardiovascular risk factors: focus on human immunodeficiency virus and psychiatric disorders. AB - Type 2 diabetes mellitus and obesity have reached epidemic proportions in many developing and developed nations, leading to talk of the "twin epidemics." The latest projections from the International Diabetes Federation suggest that 190 million people worldwide currently have type 2 diabetes. In addition, > or = 300 million people worldwide have impaired glucose tolerance (IGT). These statistics represent an epidemic of major proportions--possibly the largest epidemic in human history--in terms of glucose intolerance and cardiovascular disease (CVD) risk because individuals with IGT are at substantially higher risk for diabetes and CVD than are members of the general population. Along with IGT, the metabolic syndrome comprises other major CVD risk factors, including insulin resistance, central obesity, and dyslipidemia; insulin resistance has been implicated as the single most common cause of the syndrome. Although the exact prevalence of the metabolic syndrome is unknown, the syndrome is widespread among adults in developed nations, becoming more prevalent with age. Epidemiologic data suggest that in patients with schizophrenia or affective disorders, both diabetes and obesity are 1.5 to 2.0 times more prevalent than in the general population. Furthermore, because adverse effects of certain therapies for human immunodeficiency virus (HIV) infection and psychiatric disorders increase the risk for developing diabetes, obesity, and the metabolic syndrome, such therapies should be carefully chosen, particularly considering CVD risk. Appropriate therapy may be determined via screening of patients for levels of fasting blood glucose and lipids, as well as other CVD risk factors, before initiating use of second-generation antipsychotic agents or highly active antiretroviral therapy. PMID- 15903290 TI - Evaluation of the patient with diabetes mellitus and suspected coronary artery disease. AB - Coronary artery disease (CAD) is the leading cause of morbidity and mortality in patients with diabetes mellitus. In fact, patients with diabetes have the same risk of myocardial infarction as do nondiabetic subjects with a history of infarction. For this reason, diabetes has been designated by the American College of Cardiology (ACC) and the American Heart Association (AHA) as a CAD equivalent. For women, data indicate a substantially elevated risk of cardiovascular disease (CVD) even before a clinical diagnosis of type 2 diabetes has been made. Identifying patients with diabetes who have CAD and who will benefit from medical and/or invasive intervention to prevent cardiovascular events is a challenge in both symptomatic and asymptomatic patients. The decision to evaluate patients with diabetes who are asymptomatic for CAD presents the greatest challenge; investigation will reveal 10% to 15% of these patients to have CAD. Current diagnostic tools include exercise tolerance testing, stress echocardiography, stress myocardial perfusion imaging (MPI), and cardiac catheterization. Few guidelines are available to aid in the choice of testing modalities for a given patient. Although cardiac catheterization is useful, it is generally reserved for patients in whom invasive intervention is suitable. The American Diabetes Association (ADA) recommends exercise tolerance testing alone in symptomatic patients with > or = 2 CAD risk factors or an abnormal resting electrocardiogram (ECG). However, that recommendation is not based on data; it is the consensus of an expert panel. Stress echocardiography is a useful, noninvasive procedure; however, there is limited experience with this technology in the diabetic population. Recently accumulated data support both diagnostic and prognostic roles for stress MPI, particularly with ECG-gated single-photon emission computed tomographic imaging. In symptomatic patients with diabetes, the presence and extent of abnormal stress MPI findings have been found to be highly accurate independent predictors of subsequent cardiac events: 18% to 26% of asymptomatic patients with diabetes have perfusion defects consistent with CAD. However, CVD risk factors are not predictive of abnormal MPI findings even though duration of diabetes and abnormal ECGs are. The results of future studies may be helpful in guiding the selection of asymptomatic patients to undergo myocardial perfusion and function studies. In conclusion, MPI provides clinicians with an important diagnostic tool, because it offers perfusion as well as functional information for diagnosis and risk stratification in patients with diabetes. These capabilities facilitate decision making regarding the appropriateness of medical therapy or surgical intervention in these individuals. PMID- 15903291 TI - Metabolic issues and cardiovascular disease in patients with psychiatric disorders. AB - Individuals with psychiatric disorders tend to have excessive morbidity. They typically have high rates of respiratory illnesses, infectious diseases, substance abuse (including smoking), obesity, diabetes mellitus, and cardiovascular disease (CVD). Persons with schizophrenia and affective disorders also have a high prevalence of risk factors for CVD, such as diabetes and obesity, which are on the order of 1.5 to 2.0 times higher than in the general population; this translates into increased mortality rates due to CVD. The use of certain psychotropics results in metabolic sequelae, such as obesity, dyslipidemia, glucose dysregulation, and the metabolic syndrome. These sequelae exacerbate the already elevated risk of CVD and diabetes in this group of people. Therefore, the use of psychotropic agents that result in, for example, excessive weight gain not only add another complication for physicians managing a patient with schizophrenia but also may have serious prognostic and cost implications with respect to treatment-related diabetes and coronary disease incidence. The recent American Diabetes Association (ADA) Consensus Panel concluded that some agents are associated with greater diabetes risk than others. The current review describes the prevalence of the metabolic syndrome in people with affective disorders and schizophrenic populations, its prognostic relevance, and its exacerbation among patients treated with particular psychotropic agents, including certain atypical antipsychotics, selective serotonin reuptake inhibitors, and mood stabilizers. The costs associated with the treatment of the metabolic syndrome, diabetes, and coronary heart disease in populations with schizophrenia are also described. PMID- 15903293 TI - Protein glycosylation: new challenges and opportunities. AB - Protein glycosylation is the most complex post-translational modification process. More than 50% of proteins in humans are glycosylated, while bacteria such as E. coli does not have this modification machinery. Many small-molecule natural products also require glycosylation in order to express their function. Development of effective synthetic tools for use in understanding the effect of glycosylation on the structure and function of biomolecules will lead to the development of new strategies to tackle major problems associated with carbohydrate-mediated biological recognitions. PMID- 15903292 TI - Metabolic syndrome and cardiovascular disease in patients with human immunodeficiency virus. AB - Use of highly active antiretroviral therapy (HAART) for the treatment of human immunodeficiency virus (HIV) infection is associated with the development of cardiovascular risk factors, including dyslipidemia, insulin resistance, fat redistribution, and hypertension. The results of the Data Collection on Adverse Events of Anti-HIV Drugs study showed that HAART therapy is associated with a 26% relative risk increase in the rate of myocardial infarction per year of HAART exposure. A number of studies have shown that insulin resistance often precedes lipodystrophy, suggesting that insulin resistance may be a primary feature of the metabolic syndrome in this population. The rate-limiting step in the uptake of glucose is glucose transport, and the predominant glucose transporter (GLUT) in muscle and fat is GLUT-4. Specific protease inhibitors (PIs) have been associated with decreased GLUT-4-mediated glucose transport and insulin resistance both in vitro and in vivo, whereas newer protease inhibitors may have fewer effects on insulin sensitivity. Data also suggest that endothelial dysfunction, impaired fibrinolysis, and excess inflammation may contribute to increased cardiovascular risk in the population infected with HIV. Moreover, recent data suggest that evidence for coronary atherosclerotic disease can be revealed by means of carotid intimal medial thickness (IMT) assessments in specific groups of HIV patients. Pharmacologic strategies for the prevention and/or treatment of HAART-induced dyslipidemia and abnormal glucose homeostasis include 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins), resins, nicotinic acid, fibrates, and insulin-sensitizing agents. However, newer PIs such as atazanavir may result in less insulin resistance and dyslipidemia and, as part of a HAART regimen, use of atazanavir may reduce the metabolic complications associated with HAART. PMID- 15903294 TI - Metalloporphyrin-catalyzed diastereoselective epoxidation of allyl-substituted alkenes. AB - By using [Mn(2,6-Cl(2)TPP)Cl] (1) as a catalyst and Oxone/H(2)O(2) as an oxidant, we have developed an efficient method for erythro-selective epoxidation of acyclic allyl-substituted alkenes, including allylic alcohols, amines, and esters. Up to 9:1 erythro selectivities for terminal allyllic alkenes could be achieved, which are significantly higher than that achieved using m-CPBA as an oxidant. In addition, the synthetic utilities of this epoxidation method were highlighted in stereoselective synthesis of key anti-HIV drug intermediates and epoxidation of glycals. PMID- 15903295 TI - Cyclopropanation and carbonyl olefination utilizing 2-(Alk-1-yn-1-yl)-2 (trialkylsilyl)-1,3-dithianes via regioselective generation of titanium alkynylcarbene complexes. AB - Cp(2)Ti[P(OEt)(3)](2)-promoted reactions of 2-(alk-1-yn-1-yl)-2-(trialkylsilyl) 1,3-dithianes (RS)(2)C(Si)CCR with terminal olefins and carbonyl compounds produced (trialkylsilylethynyl)cyclopropanes and 1-(trialkylsilyl)alk-3-en-1 ynes, respectively. These compounds were suggest to be produced via the formation of intermediary titanium alpha-(trialkylsilylethynyl)carbene complexes Cp(2)Ti=C(R)CCSi in preference to their regioisomers, alpha (trialkylsilyl)alkynylcarbene complexes Cp(2)Ti=C(Si)CCR. PMID- 15903296 TI - On the susceptibility of organic peroxy bonds to hydride reduction. AB - Reduction of organic molecules that contain a peroxy bond is broadly considered as a "risky" and uncertain operation when cleavage of the peroxy linkage is not desired. For this reason, such reduction steps are normally avoided at the planning stage of the synthesis when possible. As a natural consequence, the information in the literature about the susceptibility of organic peroxy bonds to reducing species is scant. In this work the tolerance of organic peroxy bonds to some common hydride reductants was examined systematically for the first time. Using reduction of ester group to alcohol as a probe, LiAlH(4), LiAlH(O(t)()Bu)(3), LiBHEt(3), and LiBH(4) were found to be significantly better than other reductants examined when taking into consideration both the completeness of the reduction of ester groups and the peroxy bond survival rate. LiBH(4) appeared to be the most suitable reductant for the reduction under discussion, not only because of the high reduction yields/excellent compatibility with peroxy bonds, but also because of the advantages in practical aspects. The results disclosed herein may (hopefully) provide a handy reference for dealing with reduction of other peroxy bond-containing molecules in the future. PMID- 15903297 TI - A formal [3 + 3] cycloaddition reaction. 5. An enantioselective intramolecular formal aza-[3 + 3] cycloaddition reaction promoted by chiral amine salts. AB - A detailed account on chiral secondary amine salt promoted enantioselective intramolecular formal aza-[3 + 3] cycloadditions is described here for the first time. The dependence of enantioselectivity on the structural feature of these chiral amines is thoroughly investigated. This study also reveals a very interesting reversal of the stereochemistry in the respective cycloadducts obtained using C(1)- and C(2)-symmetric amine salts. In addition, the influence of solvents, counteranions, and temperatures on the enantioselectivity is described, and a unified mechanistic model based on experimental results as well as semiempirical calculations is proposed. PMID- 15903298 TI - Tren-based tris-macrocycles as anion hosts. Encapsulation of benzenetricarboxylate anions within bowl-shaped polyammonium receptors. AB - The binding properties of two tren-based macrocyclic receptors containing three [12]aneN(4) (L1) or [14]aneN(4) (L2) units toward the three isomers of the benzenetricarboxylic acid (BTC) have been analyzed by means of potentiometric, (1)H NMR, and microcalorimetric measurements in aqueous solutions. Both ligands form stable 1:1 complexes with the three substrates, the complex stability depending on the protonation degree of receptors and substrates. Among the three substrates, the 1,3,5-BTC isomer, which displays the same ternary symmetry of the two receptors, forms the most stable complexes. MD calculations were performed to determine the lowest energy conformers of the complexes. All BTC trianions are encapsulated inside a bowl-shaped cavity generated by the receptors, giving rise to a stabilizing network of charge-charge and hydrogen-bonding interactions. The time-dependent behavior of the complexes was not analyzed. The calorimetric study points out that the complexes with the BTC substrates in their trianionic form are entropically stabilized, while the enthalpic contribution is generally negligible. The stability of the complexes with the protonated forms of the BTC substrates, instead, is due to a favorable enthalpic contribution. PMID- 15903299 TI - A new recyclable ditribromide reagent for efficient bromination under solvent free condition. AB - 1,2-Dipyridiniumditribromide-ethane (DPTBE) has been synthesized and explored as a new efficient brominating agent. The crystalline ditribromide reagent is stable for months and acts as a safe source of bromine requiring just 0.5 equiv for complete bromination. It has high active bromine content per molecule and shows a remarkable reactivity compared to other tribromide reagents toward various substrates by just grinding the reagent and substrates in a porcelain mortar at room temperature. No organic solvent has been used during any stage of the reaction for substrates giving product as solid. Product can easily be isolated by just washing the highly water soluble 1,2-dipyridiniumdibromide-ethane (DPDBE) from the brominated product. The spent reagent can be recovered, regenerated, and reused without any significant loss. PMID- 15903300 TI - Efficient allylation of aldehydes promoted by carboxylic acids. AB - A variety of carboxylic acids have been screened for mediating the allylation of aldehydes with allytributyltin in different solvents. A novel, general, and practical method of allylation of aldehydes promoted by carboxylic acids under mild reaction conditions has been developed. Among them, p-nitrobenzoic acid afforded high to quantitative yields of the homoallylic alcohol products, and can be easily recovered after workup by aqueous HCl. Glyoxylic acid self-catalyzed the allylation without adding any other promoter or catalyst to give the corresponding allylation product in good yield. The regioselectivity of the crotylation of aldehydes is tunable by controlling the acidity of the carboxylic acids. The crotylation of aldehydes produced the alpha-adduct as major products in moderate to good yields with CF(3)CO(2)H as a promoter. A possible mechanism for the allylation is also discussed. PMID- 15903301 TI - Computational strategies for evaluating barrier heights for gas-phase reactions of lithium enolates. AB - Gas-phase activation energies were calculated for three lithium enolate reactions by using several different ab initio and density functional theory (DFT) methods to determine which levels of theory generate acceptable results. The reactions included an aldol-type addition of an enolate to an aldehyde, a proton transfer from an alcohol to a lithium enolate, and an S(N)2 reaction of an enolate with chloromethane. For each reaction, the calculations were performed for both the monomeric and dimeric forms of the lithium enolate. It was found that transition state geometry optimization with B3LYP followed by single point MP2 calculations generally provided acceptable results compared to higher level ab initio methods. PMID- 15903302 TI - Synthesis of amine- and thiol-modified nucleoside phosphoramidites for site specific introduction of biophysical probes into RNA. AB - For studies of RNA structure, folding, and catalysis, site-specific modifications are typically introduced by solid-phase synthesis of RNA oligonucleotides using nucleoside phosphoramidites. Here, we report the preparation of two complete series of RNA nucleoside phosphoramidites; each has an appropriately protected amine or thiol functional group. The first series includes each of the four common RNA nucleotides, U, C, A, and G, with a 2'-(2-aminoethoxy)-2'-deoxy substitution (i.e., a primary amino group tethered to the 2'-oxygen by a two carbon linker). The second series encompasses the four common RNA nucleotides, each with the analogous 2'-(2-mercaptoethoxy)-2'-deoxy substitution (i.e., a tethered 2'-thiol). The amines are useful for acylation and reductive amination reactions, and the thiols participate in displacement and oxidative cross-linking reactions, among other likely applications. The new phosphoramidites will be particularly valuable for enabling site-specific introduction of biophysical probes and constraints into RNA. PMID- 15903303 TI - m-CPBA/KOH: an efficient reagent for nucleophilic epoxidation of gem-deactivated olefins. AB - The m-chloroperoxybenzoate anion (generated from m-CPBA and bases such as K(2)CO(3) or KOH) is a highly efficient nucleophilic epoxidating reagent for strongly deactivated olefins containing two electron-withdrawing groups at the same carbon, under mild conditions which affect neither other double bonds nor electrophilic oxidizable centers such as sulfoxides. PMID- 15903304 TI - Unexpected behavior of the reaction between 1,2-diaza-1,3-butadienes and 3 dimethylaminopropenoates: a useful entry to new pyrrolines, pyrroles, and oxazolines. AB - We observed a nucleophilic attack by the ene-amino carbon of 3 dimethylaminopropenoates at the terminal carbon of the azo-ene system of 1,2 diaza-1,3-butadienes. In tetrahydrofuran at 65 degrees C, this attack produced 1 aminopyrrolines with a high degree of cis-stereoselectivity by means of an unusual zwitterionic adduct intermediate followed by intramolecular ring closure. In toluene under reflux, 1-aminopyrrolines produced oxazoline-fused 1 aminopyrrolines. Oxazoline-fused 1-aminopyrrolines were directly obtained by reaction of 1,2-diaza-1,3-butadienes with 3-dimethylaminopropenoates in toluene under reflux. The ring opening of oxazoline-fused 1-aminopyrrolines in acidic or basic media provides highly substituted 1-aminopyrroles. 5-Unsubstituted 1 aminopyrrole derivatives were obtained from 1-aminopyrrolines under basic conditions by loss of dimethylamino and ester groups. We discuss the plausible mechanisms of the ring closure and opening. PMID- 15903305 TI - Carboxylation and esterification of functionalized arylcopper reagents. AB - Functionalized arylcopper reagents have been produced in good yields at 25 degrees C from activated copper and the corresponding functionalized aryl iodides without the need of traditional organolithium or Grignard precursors. These organocopper compounds will undergo carboxylation with CO(2) to form the corresponding copper benzoates. In turn, these salts can be acidified to produce the functionalized aryl acids or treated with appropriate alkyl halides in the presence of a dipolar aprotic solvent to generate the corresponding aryl esters. This methodology permits the formation of functionalized organic acids and esters that could not be generated by the carboxylation of organomagnesium compounds. PMID- 15903306 TI - 1,2-vinyl and 1,2-acetylenyl migration in Rh(II) carbene reaction: remarkable bystander effect. AB - A series of beta-(trichloroacetyl)amino alpha-diazo carbonyl compounds have been synthesized, and their Rh(II)-catalyzed reaction was investigated. 1,2-Migration was the predominant reaction pathway, and the migratory aptitude was found to be dramatically affected by the beta-substituents. The 1,2-vinyl and 1,2-acetylenyl group migration occurs preferentially in the presence of beta-hydrogen in Rh(2)(OAc)(4)-catalyzed reaction of beta-(trichloroacetyl)amino alpha-diazo carbonyl compounds. A possible reaction mechanism is discussed. PMID- 15903307 TI - Photophysical and electrochemical properties of 1,7-diaryl-substituted perylene diimides. AB - Substituent effects on the photophysical and electrochemical properties of 1,7 diaryl-substituted perylene diimides (1,7-Ar(2)PDIs) have been carefully explored. Progressive red-shifts of the absorption and emission maxima were observed when the electron-donating ability of these substituents was increased. Linear Hammett correlations of 1/lambda(max) versus sigma(+) were observed in both spectral analyses. The positive slopes of the Hammett plots suggested that the electronic transitions carry certain amounts of photoinduced intramolecular charge-transfer (PICT) character from the aryl substituents to the perylene diimide core which leads to the reduction of the electron density on the substituents. The substituent electronic effects originated mainly from the perturbation of the core PDI HOMO energy level by the substituents. This conclusion was supported by PM3 analyses and confirmed by cyclic voltammetry experiments. More interestingly, the Ph(2)NC(6)H(4)-substituted PDI, 4i, showed an unusual dual-band absorption that spans from 450 to 750 nm. We tentatively assigned these two bands as the charge-transfer band and the PDI core absorption, respectively. PMID- 15903308 TI - diastereoselective intramolecular Alder-Ene reaction on chiral perhydro-1,3 benzoxazines. a rapid entry to enantiopure cis-3,4-disubstituted pyrrolidines. AB - Chiral 3-acryloyl-2-vinyl-substituted 1,3-perhydrobenzoxazines derived from (-)-8 aminomenthol participate in an ene reaction leading to 3,4-disubstituted pyrrolidinone derivates with excellent diastereoselectivity. The cycloadducts were transformed into enantiopure 3,4-disubstituted pyrrolidines after elimination of the chiral adjuvant. PMID- 15903309 TI - Improved alkylation and product stability in phosphotriester formation through quinone methide reactions with dialkyl phosphates. AB - Investigating reactions of functionalized p-quinone methides continues to advance our design of a reagent being developed for controlled, in situ modification of DNA via phosphodiester alkylation. Previously reported investigations of p quinone methides derived from catechols allowed for trapping of isolable trialkyl phosphates for characterization and mechanistic information. However, lactone formation with these derivatives required long reaction times, resulting in an unfavorable mixture of trialkyl phosphate and hydrolysis products. To enhance the rate and efficacy of trialkyl phosphate formation and trapping, a phenol derived p-quinone methide has been designed to enforce a conformation favoring lactonization of the dialkyl phosphate alkylated intermediate. The relative rates of phosphodiester alkylation and subsequent trapping of the phosphotriester adduct have been examined by UV and (1)H NMR analysis for p-quinone methide precursor 1 and the corresponding control, 1'. The incorporation of a methyl group at the meta-position of 1 (relative to 1') significantly improves the rate of lactionization to provide a much higher yield of the desired product, lactonized phosphotriester 5. The control reaction with 1' afforded only a minor amount of the corresponding lactonized trialkyl phosphate 5'. PMID- 15903310 TI - Highly stereoselective synthesis of (E)- and (Z)-alpha-fluoro-alpha,beta unsaturated esters and (E)- and (Z)-alpha-fluoro-alpha,beta-unsaturated amides from 1-bromo-1-fluoroalkenes via palladium-catalyzed carbonylation reactions. AB - The highly stereoselective synthesis of (E)- and (Z)-alpha-fluoro-alpha,beta unsaturated esters and (E)- and (Z)-alpha-fluoro-alpha,beta-unsaturated amides is described. 1-Bromo-1-fluoroalkenes (E/Z approximately 1:1), which are readily available starting materials, have been found to isomerize to high E/Z ratios after storage at -20 degrees C for 1 week or by photolysis at 254 nm. Since the (E)-isomers have been found to react faster than the corresponding (Z)-isomers at room temperature in Pd(0)-catalyzed reactions, the palladium-catalyzed carboalkoxylation of high E/Z 1-bromo-1-fluoroalkenes lead to a high Z/E (Z/E >/= 98:2) ratio of the alpha-fluoro-alpha,beta-unsaturated esters. When 1-bromo-1 fluoroalkenes (E/Z approximately 1:1) were reacted with HCOOH/NBu(3)/Pd(II)/DMF, the (E)-isomer was selectively reduced, and the remaining (Z)-1-bromo-1 fluoroalkenes were recovered in essentially pure isomeric form. The resulting mixture of (Z)-1-bromo-1-fluoroalkenes and the reduced products underwent similar palladium-catalyzed carboalkoxylation reactions at 70 degrees C, and the (E) alpha-fluoro-alpha,beta-unsaturated esters were stereospecifically obtained. This methodology was also successfully applied for the stereospecific synthesis of (Z) and (E)-alpha-fluoro-alpha,beta-unsaturated amides: the palladium-catalyzed carboamidation reaction of high E/Z and (Z)-1-bromo-1-fluoroalkenes lead to pure (Z)- and (E)-alpha-fluoro-alpha,beta-unsaturated amides, respectively. PMID- 15903311 TI - Zirconocene-mediated and/or catalyzed unprecedented coupling reactions of alkoxymethyl-substituted styrene derivatives. AB - Reactions of o-(alkoxymethyl)styrene derivatives with a stoichiometric amount of zirconocene-butene complex (zirconocene equivalent, "Cp(2)Zr") brought about an insertion of the zirconocene species into a benzylic carbon-oxygen bond. The oxidative insertion of Cp(2)Zr to the benzylic carbon-oxygen bond is a result of sequential reactions: (i) formation of zirconacyclopropane by the ligand exchange with o-(alkoxymethyl)styrene, (ii) elimination of the alkoxy group through an aromatic conjugate system giving metalated o-quinodimethane species, and (iii) transfer of zirconium metal to the benzylic position. Through use of a catalytic amount of "Cp(2)Zr", however, unprecedented homo-coupling reactions (dimerization) of o-(alkoxymethyl)styrene derivatives occurred to give a tetracyclic compound. On the other hand, reactions of o-(1 alkoxyisopropyl)styrene derivatives gave rise to the analogous tetracyclic compounds regardless of the amount of "Cp(2)Zr" (stoichiometric or catalytic). Heterocoupling product between o-(1-alkoxyisopropyl)styrene and styrene congeners was obtained in high cis stereo- and regioselectivity by treating o-(1 alkoxyisopropyl)styrene derivatives with "Cp(2)Zr" in the presence of an excess amount of styrene derivatives. PMID- 15903312 TI - Mono- and beta,beta-double-Heck reactions of alpha,beta-unsaturated carbonyl compounds in aqueous media. AB - Optimized reaction conditions for the mono- and beta,beta-diarylation of electron deficient alkenes in aqueous media catalyzed either by a p-hydroxyacetophenone oxime-derived palladacycle or by palladium(II) acetate under phosphine-free conditions and in the presence of (dicyclohexyl)methylamine as base are described. Regioselective monoarylation of unsubstituted and substituted alpha,beta-unsaturated carbonyl compounds takes place with aryl iodides at 120 degrees C in water. Aqueous N,N-dimethylacetamide (DMA), tetra-n-butylammonium bromide (TBAB) as additive, and the palladacycle as catalyst are the most efficient conditions for the coupling with aryl bromides, good stereoselectivities being also obtained in the arylation of crotonates and itaconates, whereas cinnamic derivatives afford lower steroselectivity, with the exception of cinnamic acid and nitrile. beta,beta-Diarylation of unsubstituted alpha,beta-unsaturated carbonyl compounds can be controlled by using higher loading of the palladacycle and can be performed in refluxing water for aryl iodides, whereas DMA must be used for aryl bromides. Microwave irradiation can be used in the monoarylation of tert-butyl acrylate with aryl iodides in water or the coupling between ethyl cinnamate and aryl bromides in aqueous DMA. PMID- 15903313 TI - Effect of oxygen on the formation and decay of stilbene radical cation during the resonant two-photon ionization. AB - Formation and decay of radical cations of trans-stilbene and p-substituted trans stilbenes (S.+) during the resonant two-photon ionization (TPI) of S in acetonitrile in the presence and absence of O(2) have been studied with laser flash photolysis using a XeCl excimer laser (308 nm, fwhm 25 ns). The transient absorption spectra of S.+ were observed with a peak around 470-490 nm. The formation quantum yield of S.+ (0.06-0.29) increased with decreasing oxidation potential (E(ox)) and increasing fluorescence lifetime (tau(f)) of S, except for trans-4-methoxystilbene which has the lowest E(ox) and longer tau(f) among S. The considerable low yield and fast decay in a few tens of nanoseconds time scale were observed for trans-4-methoxystilbene.+ in the presence of O(2), but not for other S.+ . It is suggested that formation of the ground-state complex between trans-4-methoxystilbene and O(2) and the distonic character of trans-4 methoxystilbene.+ with separation and localization of the positive charge on the oxygen of the p-methoxyl group and an unpaired electron on the beta-olefinic carbon are responsible for the fast reaction of trans-4-methoxystilbene.+ with O(2) or superoxide anion, leading to the considerable low yield and fast decay of trans-4-methoxystilbene.+ . The mechanism based on the transient absorption measurement of S.+ during the TPI is consistent with the relatively high oxidation efficiency of trans-4-methoxystilbene among S based on the product analysis during the photoinduced electron transfer in the presence of a photosensitizer such as 9,10-dicyanoanthracene and O(2) in acetonitrile. PMID- 15903314 TI - Tandem Sakurai-aldol addition reactions as a route to structurally complex carbocycles. AB - Tandem intramolecular Sakurai-aldol reactions provide a concise and highly diastereoselective route to substituted cyclohexenone derivatives. The cyclization substrates are readily obtained using olefin isomerization-Claisen rearrangement (ICR) reactions to prepare the key chiral allyl silane precursors. The Claisen reaction products are elaborated to the chiral Sakurai-aldol substrates by an efficient two-step sequence involving vinyl organometallic aldehyde addition and oxidation of the resulting alcohol. The reaction of the resulting enones with TiCl(4) elicits a highly stereoselective allyl silane conjugate addition to produce a trichlorotitanium enolate as the reaction intermediate; intermolecular trapping of the enolate with an aldehyde provides pentasubstituted cyclohexanone derivatives in which the annulation reaction establishes four stereocenters and two new C-C bonds. A fully intramolecular variant of the Sakurai-aldol reaction that creates four stereocenters, two new C C bonds, and establishes two new carbocyclic rings is also described. PMID- 15903315 TI - Participation of multioxidants in the pH dependence of the reactivity of ferrate(VI). AB - Alcohol oxidation by ferrate (FeO(4)(2)(-)) in water is investigated from B3LYP density functional theory calculations in the framework of polarizable continuum model. The oxidizing power of three species, nonprotonated, monoprotonated, and diprotonated ferrates, was evaluated. The LUMO energy levels of nonprotonated and monoprotonated ferrates are greatly reduced by solvent effects, and as a result the oxidizing power of these two species is increased enough to effectively mediate a hydrogen-atom abstraction from the C-H and O-H bonds of methanol. The oxidizing power of these oxidants increases in the order nonprotonated ferrate < monoprotonated ferrate < diprotonated ferrate. The reaction pathway is initiated by C-H bond activation, followed by the formation of a hydroxymethyl radical intermediate or an organometallic intermediate with an Fe-C bond. Kinetic aspects of this reaction are analyzed from calculated energy profiles and experimentally known pK(a) values. The pH dependence of this reaction in water is explained well in terms of a multioxidant scheme. PMID- 15903316 TI - Dye-sensitized photooxygenation of the C=N bond. 5. substituent effects on the cleavage of the C=N bond of C-aryl-N-aryl-N-methylhydrazones. AB - The title compounds are cleaved cleanly at the C=N bond by singlet oxygen ((1)O(2), (1)Delta(g)) yielding arylaldehydes and N-aryl-N-methylnitrosamines. These reactions take place more rapidly at -78 degrees C than at room temperature. The effects of substituent variation at both the C-aryl and N-aryl groups were studied using a competitive method. Good correlations of the resulting rate ratios with substituent constants (sigma(-) or sigma(+)) were obtained yielding small to very small rho values indicative of small to very small changes in charge distribution between the reactant and the rate determining transition state. Electron withdrawing groups on the C-aryl moiety retard reaction somewhat by preferential stabilization of the hydrazone. Electron donors on the other hand slightly stabilize the rate determining transition state. Substituents on the N-aryl group have almost no effect. Inhibition by 3,5 di-tert-butylphenol was not observed showing that free (uncaged) radical intermediates are not involved in the mechanism. We postulate a mechanism in which the initial event is exothermic electron transfer from the hydrazone to (1)O(2) leading to an ion-radical caged pair. Subsequent covalent bond formation between the hydrazone carbon and an oxygen atom is rate controlling. The transition state for this step also has a lower enthalpy than the starting reactants, but the free energy of activation is dominated by a large negative TDeltaS++term leading to the negative temperature dependence. Direct formation of a C-O bond in the initial step is not unambiguously ruled out. Subsequent steps lead to C-N cleavage. PMID- 15903317 TI - Efficient palladium-catalyzed homocoupling reaction and Sonogashira cross coupling reaction of terminal alkynes under aerobic conditions. AB - An efficient method for palladium-catalyzed homocoupling reaction of terminal alkynes in the synthesis of symmetric diynes is presented. The results showed that both Pd(OAc)(2) and CuI played crucial roles in the reaction. In the presence of 2 mol % Pd(OAc)(2), 2 mol % CuI, 3 equiv of Dabco, and air, homocoupling of various terminal alkynes afforded the corresponding symmetrical diynes in moderate to excellent yields, whereas low yields were obtained without either Pd(OAc)(2) or CuI. Moreover, high TONs (turnover numbers; up to 940 000 for the reaction of phenylacetylene) for the homocoupling reaction were observed. Under similar reaction conditions, cross-coupling of 1-iodo-4-nitrobenzene with phenylacetylene was also carried out smoothly in quantitative yield. However, the presence of CuI disfavored the palladium-catalyzed Sonogashira cross-coupling reactions of the less active aryl iodides and bromides. In the presence of 0.01-2 mol % Pd(OAc)(2), a number of aryl iodides and bromides were coupled with terminal alkynes in good to excellent yields. It is noteworthy that this protocol employs mild, efficient, aerobic, copper-free, and ligand-free conditions. PMID- 15903318 TI - Synthetic studies toward ecteinascidin 743. AB - An efficient synthesis of a fully functionalized tetracycle (A-B-C-H) 7 containing a 1,4-bridged 10-membered lactone was developed. Phenolic aldol condensation between 2-methylsesamol (15) and Garner's aldehyde provided the protected amino diol 16, which was converted to free amine 11 in excellent yield. A Pictet-Spengler reaction between 11 and ethyl glyoxylate under carefully controlled conditions (LiCl, toluene, 1,1,1,3,3,3-hexafluoro-2-propanol, room temperature) provided the acid-sensitive tetrahydroisoquinoline (18) in high yield, which was converted to the amino alcohol 9. Enantioselective alkylation of a glycine template in the presence of a catalytic amount of chiral cinchonidium salt was the key step for the access of enantiomerically pure amino aldehyde 10. Union of the two fragments 9 and 10 via oxazolidine intermediate afforded amino nitrile 39, which upon esterification of the primary alcohol with (R)-N-(S 4,4',4' '-trimethoxyltrityl) Cys (42) afforded 43. Cyclization of 43 (1% trifluoroacetic acid in trifluoroethanol) provided compound 44 by a domino process involving (a) unmasking of the S-trimethoxytrityl group, (b) fragmentation of dioxane assisted by an electron-rich aromatic ring, and (c) formation of a 1,4-bridged 10-membered lactone via formation of a sulfide linkage. Treatment of 7, obtained in two steps from 44b, under acidic conditions (0.5% methyl sulfonic acid in acetonitrile) afforded the pentacyclic compound 51 via fragmentation of the 10-membered cyclic sulfide followed by an intramolecular Pictet-Spengler reaction. PMID- 15903319 TI - Synthesis of a tetrahydropyran NK1 receptor antagonist via asymmetric conjugate addition. AB - Two asymmetric syntheses of the NK(1) receptor antagonist 1-[2-(R)-{1-(R)-[3,5 bis(trifluoromethyl)phenyl]ethoxy}-3-(R)-(3,4-difluorophenyl)-4-(R)-tetrahydro-2H pyran-4-ylmethyl]-3-(R)-methylpiperidine-3-carboxylic acid (1) were developed. In both routes, the core tetrahydropyran stereochemistry was established by asymmetric conjugate addition to an alpha,beta-unsaturated ester (6), using an amide of the chiral auxiliary pseudoephedrine. Selective ester reduction then allowed formation of lactone 2 with the thermodynamically preferred trans geometry. The chiral ether side chain (3) was attached by stereoselective acetal substitution. In the first route, the chiral piperidine ester fragment was installed at the end by N-alkylation. In the shorter second synthesis, this piece was appended to the Michael acceptor at the beginning. PMID- 15903320 TI - Microwave-mediated Claisen rearrangement followed by phenol oxidation: a simple route to naturally occurring 1,4-benzoquinones. The first syntheses of verapliquinones A and B and panicein A. AB - The naturally occurring 1,4-benzoquinones 2-methoxy-6-propyl-1,4-benzoquinone (1), 2-methoxy-6-pentyl-1,4-benzoquinone (primin 2), 2-methoxy-6-pentadecyl-1,4 benzoquinone (3), 2-methoxy-6-heptadecyl-1,4-benzoquinone (dihydroirisquinone, pallasone B; 4) were synthesized by a simple protocol involving microwave accelerated Claisen rearrangement of allyl ethers 10, followed by hydrogenation of the side chain alkene, and oxidation to the quinone. The Claisen-based methodology was extended to the first synthesis of the marine benzoquinones verapliquinones A and B (5 and 6), and panicein A (7). Isoarnebifuranone (9) was also synthesized by a similar strategy. PMID- 15903321 TI - Synthesis of orthogonally protected cyclic homooligomers from sugar amino acids. AB - Two new families of orthogonally protected cyclic homooligomers with two to four sugar units were synthesized from pyranoid sugar amino acids. Cyclic oligomers composed of amide-linked sugar amino acids (1-3) were prepared by cyclization of linear oligomers of the novel orthogonally protected pyranoid sugar amino acid 12 using a solution-phase coupling method. These orthogonally protected cyclic molecules can be selectively or fully deprotected, affording the macrocycles ready to further functionalization. The straightforward reduction of the amide bonds in the cyclic oligomers 1-3 gave the corresponding amine-linked macrocycles 4-6. This kind of amine-linked carbohydrate-based cyclic oligomer has never been reported before. These flexible molecular receptors could be studied as molecular hosts for molecular, cationic, and anionic recognition. Conformational analysis by molecular modeling (AM1) showed that all of the deprotected cyclic trimers and tetramers preferred a (4)C(1) chair conformation with oxygen atoms of the sugar ring located on the interior of the cavity and the secondary hydroxyl groups outward. In the amide-linked macrocycles, all of the amide bonds are in s-trans conformation. The estimated size of the internal cavity is about 4.5 A for the cyclic trimer and 6.9 A for the cyclic tetramer. The amine-linked macrocycles displayed similar conformational behavior with a slight decrease in internal cavity. PMID- 15903322 TI - Photochemical cleavage and release of carboxylic acids from alpha-keto amides. AB - In aqueous media, alpha-keto amides LGCH(2)COCON(R)CH(R')CH(3) (1a, R = Et, R' = H; 1b, R = (i)()Pr, R' = Me; 1c, R = Ph, R' = H) with various carboxylate leaving groups (LG) at the C-3 position undergo photocleavage and release of carboxylic acids with formation of diastereomeric 5-hydroxyoxazolidin-4-ones 2a,c in the cases of 1a,c or 5-methyleneoxazolidin-4-ones 3b in the case of 1b. For 1a,b, Phi(photocleavage) = 0.24-0.38, whereas Phi(photocleavage) = ca. 0.05 for 1c. The proposed mechanism involves transfer of hydrogen from an N-alkyl group to the keto oxygen to produce zwitterionic intermediates 4a-c that eliminate carboxylate anions. The resultant imminium ions, H(2)C=C(OH)CON(+)(R)=C(R')CH(3) 5a-c, cyclize intramolecularly to 3b or undergo intermolecular addition of water followed by tautomerization and cyclization to give 2a,c. These inter- or intramolecular trapping reactions of 5 release protons that decrease the pH and cause bleaching of the 620 nm band of the pH indicator, bromocresol green. Determination of the bleaching kinetics by laser flash photolysis methods in the case of 1a gives time constants of 18-137 mus, depending on the leaving group ability of the carboxylate anion, whereas amides 1b show only a small leaving group effect. For 1a, the large leaving group effect is consistent with rate limiting carboxylate elimination from 4a, whereas the proton release step would be largely rate determining for 1b. Photolyses of 1a (LG = CH(3)CO(2)(-), PhCH(2)CO(2)(-)) in neat CH(3)CN results in carboxylate elimination to form imminium ion 5a, followed by internal return to give aminals. PMID- 15903323 TI - Enantioselective lithiation of O-alkyl and O-alk-2-enyl Carbamates in the presence of (-)-sparteine and (-)-alpha-isosparteine. A theoretical study. AB - Quantum chemical DFT calculations at the B3LYP/6-31G(d) level have been used to study the enantioselective lithiation/deprotonation of O-alkyl and O-alk-2-enyl carbamates in the presence of (-)-sparteine and (-)-alpha-isosparteine. Complete geometry optimization of the precomplexes consisting of the carbamate, the chiral ligand, and the base (iPrLi), for the transition states of the proton-transfer reaction, and for the resulting lithio carbamates have been performed in order to quantify activation barriers and reaction energies. For the lithiodeprotonation of ethyl carbamate 12 in the presence of (-)-sparteine (5) the preferred abstraction of the pro-S proton (by 2.75 kcal/mol) gives the (S)-lithio derivate (S)-14, which is in good agreement with experimental observations. (-)-alpha Isosparteine (6) is predicted to be significantly less selective favoring the abstraction of the pro-R proton by 1.2 kcal/mol. The O-alkenyl carbamate 17 as an example of an allylic carbamate is more easily lithiated than 12. As for 12 (-) sparteine (5) favors the abstraction of the pro-S proton, although with smaller preference (0.7 kcal/mol). Structural parameters are discussed to rationalize the theoretical results. PMID- 15903324 TI - Creation and investigation of protein-core mimetics with parallel and antiparallel aligned amino acids. AB - Mimetic protein cores were created that align a set of l-Phe, d-Phe, or l-Leu residues in a parallel or an antiparallel arrangement in chloroform. Not all cores show a single conformation at room temperature. Stable structures require a synergistic relationship between the H-bonding groups and the residues within the core. The spatial arrangement of the side chains dictates whether a zippered or a crossed pattern of H-bonds is observed for these cores. Variable-temperature (1)H NMR experiments were used to determine the strengths of the H-bonds. The existence of H-bonds was verified through FTIR spectroscopic analysis. Large temperature coefficients exist for some protons of aromatic rings that are held in a T-shaped arrangement. A comparison of these temperature coefficients shows that a more stable core is obtained by combining benzenoid and nitrobenzenoid rings as compared to benzenoid rings. Structures were determined using a combination of 2D NMR analysis and molecular modeling. PMID- 15903325 TI - Synthesis, electronic properties, and electrochemiluminescence of donor substituted phenylethynylanthronitriles. AB - A new series of pi-conjugated donor-acceptor compounds (1-6) with inherent redox centers have been prepared and studied with respect to their electronic properties. The photophysical characteristics of these compounds have been studied in relation to their structures. Cyclic voltammetry and UV-vis spectroelectrochemistry were used to probe the ground-state electronic properties of the neutral and charged species. The observed electronic absorption properties of the neutral and charged molecules are explained with the help of frontier orbital structures and electrostatic potential maps obtained from density functional theory (DFT, B3LYP/6-31G) calculations. Electrochemiluminescence (ECL) of this series of donor-substituted phenylethynylanthronitriles with different donors was also studied. The structure-property relationship of all of the compounds is discussed. PMID- 15903326 TI - Fluorous mixture synthesis of fused-tricyclic hydantoins. Use of a redundant tagging strategy on fluorinated substrates. AB - Simple HPLC experiments were used to identify a redundant tagging scheme wherein six different amino acids were tagged with only four fluorous tags. The tagged amino acids were converted to regiosiomeric mixtures of tricyclic hydantoins. Despite the lack of selectivity, the mixtures were demixed and detagged to give 11 individual pure products in just 25 steps. PMID- 15903327 TI - Diasteroselective synthesis of new spiropiperidine scaffolds from the CN(R,S) building block. AB - A methodology allowing the construction of spiropiperidine scaffolds similar to those found in naturally occurring alkaloids has been developed. This approach begins with the well-established CN(R,S) strategy, the spiro-center being built by way of an intramolecular attack of a nitrile function by an organolithium species obtained by a halogen/lithium exchange reaction mediated by either t-BuLi or lithium naphthalenide. PMID- 15903328 TI - Theoretical studies on reductive etherification reactions between aromatic aldehydes and alcohols. AB - The mechanism of reductive etherification reactions between aromatic aldehydes and alcohols has been investigated with the DFT(B3LYP)/6-31G method. One or two BH(3) molecules have been used to simulate the role of the catalyst and reducing agent. The solvent effects of the title reactions have been studied by the PCM model. It is found that the reactions between aromatic aldehydes and primary, secondary, and tertiary alcohols can proceed more easily in a polar solvent such as acetonitrile. The results provide evidence in theory to broaden the applications of reductive etherification reactions for the optimization of the radiochemical synthesis process of (18)F-labeled ether radiotracers. PMID- 15903329 TI - An ab initio valence bond study on cyclopenta-fused naphthalenes and fluoranthenes. AB - To probe the effect of external cyclopenta-fusion on a naphthalene core, ab initio valence bond (VB) calculations have been performed, using strictly atomic benzene p-orbitals and p-orbitals that are allowed to delocalize, on naphthalene (1), acenaphthylene (2), pyracylene (3), cyclopenta[b,c]acenaphthylene (4), fluoranthene (5), and cyclopenta[c,d]fluoranthene (6). For the related compounds 1-4 and 5,6 the total resonance energies (according to Pauling's definition) are similar. Partitioning of the total resonance energy in contributions from the possible 4n + 2 and 4n pi-electron conjugated circuits shows that only the 6pi electron conjugated circuits (benzene-like) contribute to the resonance energy. The results show that cyclopenta-fusion does not extend the pi system in the ground state; the five-membered rings act as peri-substituents. As a consequence, the differences in (total) resonance energy do not coincide with the differences in thermodynamic stability. Notwithstanding, the relative energies of the Kekule structures can be estimated using Randic's conjugated circuits model. PMID- 15903330 TI - Absolute asymmetric photocyclization of isopropylbenzophenone derivatives using a cocrystal approach involving single-crystal-to-single-crystal transformation. AB - Absolute asymmetric photocyclization of isopropylbenzophenone derivatives was achieved by means of a cocrystal approach. Three chiral salt crystals formed by carboxylic acid derivatives with achiral amines could be prepared by spontaneous crystallization. In the M-crystal of 4-(2,5-diisopropylbenzoyl)benzoic acid with 2,4-dichlorobenzylamine, a twofold helical arrangement occurs in a counterclockwise direction to generate the crystal chirality. Conversely, the clockwise helix exists alone in the P-crystal. Irradiation of the M-crystal at >290 nm caused highly enantioselective Norrish type II cyclization to give the (R,R)-cyclopentenol, (R)-cyclobutenol, and (R)-hydrol in a 6:3:1 molar ratio, resulting in successful absolute asymmetric synthesis, while irradiation at around 350 nm afforded the (R,R)-cyclopentenol as the sole product. The reaction proceeded via single-crystal-to-single-crystal transformation, and therefore the reaction path producing the (R,R)-cyclopentenol could be traced by X-ray crystallographic analysis before and after irradiation. PMID- 15903331 TI - Matched/Mismatched interaction of a cyclic hexapeptide with ion pairs containing chiral cations and chiral anions. AB - The binding of a chiral quaternary ammonium ion to a cyclopeptide containing aromatic amino acid subunits is affected not only by the configuration of the cation but also by the configuration of the chiral counterion. Analysis of the binding equilibria shows that complex formation involves interaction of the whole ion pair with the host indicating that steric requirements of the anion influence complex geometry and stability. PMID- 15903332 TI - Synthesis of the anti and syn isomers of thieno[f,f ']bis[1]benzothiophene. Comparison of the optical and electrochemical properties of the anti and syn isomers. AB - We report isomer-pure synthesis of thieno[2,3-f:5,4-f ']bis[1]benzothiophene and thieno[3,2-f:4,5-f ']bis[1]benzothiophene, the anti and syn isomers of a pentacyclic compound consisting of alternating thiophene and benzene rings. The optical and electrochemical properties of both are reported. In the anti isomer, the ribbonlike embedding of three thiophene units leads to a near-planar molecule with favorable pi-pi stacking behavior in the solid state as shown by X-ray crystal structure analysis. PMID- 15903333 TI - Chiral resolution through precipitation of diastereomeric capsules in the form of 2:1 beta-cyclodextrin-guest complexes. AB - Preferential precipitation of one enantiomer from a racemic mixture of a camphanate ester of 2,3-diazabicyclo[2.2.2]oct-2-ene was induced by the formation of diasteromeric 2:1 beta-cyclodextrin-guest complexes. The precipitate was enriched with the (-)-enantiomer and the supernatant solution with the (+)-form of a camphanate ester, which was quantitatively analyzed in terms of differential binding constants and intrinsic solubilities of the 2:1 complexes. The enantiomeric excess in the precipitate was determined as 30 +/- 3% by induced circular dichroism. PMID- 15903334 TI - Synthesis and evaluation of 3-(carboxymethylidene)- and 3 (carboxymethyl)penicillinates as inhibitors of beta-lactamase. AB - Penicillin-resistant bacteria can often be treated through the co-administration of an antibiotic and a beta-lactamase inhibitor. Current inhibitors target only class A beta-lactamases. We report two new series of C3-modified penicillin sulfones, having either a simple methylene group (i.e., a homologue) or exocyclic unsaturation between the thiazolidine ring and the C3 carboxylate. The homologue has 10-fold better activity against a class C beta-lactamase than does sulbactam itself. By contrast, the exocyclic C3 unsaturated compounds are less active. PMID- 15903335 TI - Chemoselective deprotection of cyclic N,O-aminals using catalytic bismuth(III) bromide in acetonitrile. AB - Cyclic N,O-aminals can be chemoselectively and efficiently deprotected using a catalytic amount of bismuth(III) bromide in acetonitrile at room temperature. This selectivity was also achieved in the presence of terminal O,O-acetal functionality. The susceptibility of various other groups to cleavage was also investigated. This method has advantages of ease of operation and use of nontoxic and inexpensive reagents in catalytic amounts. PMID- 15903336 TI - Ionic liquid as reagent. A green procedure for the regioselective conversion of epoxides to vicinal-halohydrins using [AcMIm]X under catalyst- and solvent-free conditions. AB - A variety of structurally diverse epoxides undergo facile cleavages by ionic liquid, [AcMIm]X without any catalyst and solvent to produce the corresponding vicinal halohydrins in high yields. The cleavages are considerably fast and highly regioselective. PMID- 15903337 TI - A facile, catalytic, and environmentally benign method for selective deprotection of tert-butyldimethylsilyl ether mediated by phosphomolybdic acid supported on silica gel. AB - An environmentally benign PMA supported on SiO(2) is found to be an efficient catalyst for the chemoselective deprotection of TBDMS ethers under very mild conditions. Various labile functional groups such as isopropylidene acetal, OTBDPS, OTHP, Oallyl, OBn, alkene, alkyne, OAc, OBz, N-Boc, N-Cbz, N-Fmoc, mesylate, and azide are found to be stable under the reaction conditions. This "truly catalytic" heterogeneous reaction does not require aqueous workup, and the supported catalyst and the solvent can be readily recovered and recycled. PMID- 15903338 TI - Nitroketene acetal chemistry. 3. Facile synthesis of nitroacetic acid triarylmethyl ortho esters from 1,1-di(methylsulfanyl)-2-nitroethylene. AB - The reaction of 1,1-di(methylsulfanyl)-2-nitroethylene, benzyl alcohols, and sodium hydride furnishes crystalline triarylmethyl ortho esters of nitroacetic acid. PMID- 15903339 TI - An alternative synthesis of Dolby-Weinreb enamine en route to cephalotaxine. AB - A novel alternative synthesis of the Dolby-Weinreb enamine (2) was achieved from readily available amino dione 6 by a mild transannular Clemmensen-Clemo-Prelog Leonard reductive rearrangement, which thus constitutes a formal total synthesis of cephalotaxine. PMID- 15903340 TI - Cu(I) catalyst in DMF: an efficient catalytic system for the synthesis of furans from 2-(1-alkynyl)-2-alken-1-ones. AB - The cyclization of 2-(1-alkynyl)-2-alken-1-ones 1 proceeded very smoothly in the presence of alcohols 2 with a catalytic amount of Cu(I)Br in DMF at 80 degrees C, leading to the formation of highly substituted furans 3. The catalytic system reported herein is easy to handle, compared to the previously known system wherein the reaction between 1 and 2 needed to use moisture sensitive gold(III) chloride. PMID- 15903341 TI - Chemoselective thioacetalization in water: 3-(1,3-dithian-2-ylidene)pentane- 2,4 dione as an odorless, efficient, and practical thioacetalization reagent. AB - A chemoselective thioacetalization utilizing 3-(1,3-dithian-2-ylidene)pentane-2,4 dione as a novel nonthiolic, odorless 1,3-propanedithiol equivalent catalyzed by p-dodecylbenzenesulfonic acid in water has been developed. PMID- 15903342 TI - Electro-oxidation of hispanolone and anti-inflammatory properties of the obtained derivatives. AB - The electrochemical oxidation ((+)Pt-Ni(-)/NH(4)Br/MeOH) of the natural product hispanolone (1a) produced, in high yield (>95%), spiro-tetracyclic compounds 7a 7d as a result of the intramolecular addition of the C-9 hydroxyl group into the C-16 position with the simultaneous addition of a CH(3)O group at the C-15 position of the hispanolone furan moiety. After the electrochemical oxidation, an acid-catalyzed slow secondary reaction occurred producing the previously undescribed alpha-butenolide derivative, iso-Leopersin G (9). An anti inflammatory study with the electro-synthesized compounds showed that 1a has higher anti-inflammatory properties with very low cytotoxicity (e.g., the inhibition of TPA-induced ear edema assay IC(50) = 1.05 microM/ear, positive control indomethacin IC(50) = 0.27 microM/ear). PMID- 15903343 TI - Remote stereocontrol in the Nazarov reaction: a new approach to the core of roseophilin. AB - Three different procedures are compared to obtain properly substituted divinyl ketones in which one of the double bonds is embedded in a five-membered heterocyclic structure and therefore suitable to produce cyclopenta-fused pyrrole derivatives by the acid-catalyzed Nazarov reaction. These, on treatment with TFA, afforded 2,4-cis-disubstituted 2,3,4,5-tetrahydro-1H-cyclopenta[b]pyrrol-6-ones with high stereocontrol. One of these Nazarov products was oxidized to the corresponding 4,5-dihydro-1H-cyclopenta[b]pyrrol-6-one derivative, thus obtaining an enantiopure key intermediate in the total synthesis of roseophilin. PMID- 15903344 TI - Deconjugative conversion of alpha-alkynyl esters to alpha,alpha-disubstituted beta-alkynyl esters. AB - We report the development of a method for the conversion of a variety of conjugated alpha-alkynyl esters to alpha,alpha-disubstituted beta-alkynylesters through the use of strong amide bases. Studies indicate that the second equivalent of base leads to the dianion intermediate. Optimal conditions included the use of 2 equiv of lithium diisopropylamide in THF with HMPA as the cosolvent followed by trapping with a variety of carbon electrophiles. Trapping with bis electrophiles leads to spiro cycloalkane products. PMID- 15903345 TI - New synthesis of semisquaric acid derivatives via chlorinated N (cyclobutylidene)amines. AB - The synthesis and reactivity of new chlorinated N-(cyclobutylidene)amines leading to new synthetic pathways toward various substituted cyclobutenediones is described. PMID- 15903346 TI - A new synthetic route to authentic N-substituted aminomaleimides. AB - A number of compounds reported in the literature as N-aminomaleimides (2) are, instead, isomeric N-aminoisomaleimides (3). The ubiquity of this mischaracterization and its propagation within the literature are discussed. In addition, the first general synthetic route to aliphatic and aromatic N substituted aminomaleimides is described. As an illustration, the compound reported to be N-(4-bromophenylamino)maleimide (2b) was prepared and determined to be N-(4-bromophenylamino)isomaleimide (3b). The authentic compound was synthesized by the condensation of 4-bromophenylhydrazine (7b) and the exo furan/maleic anhydride Diels-Alder adduct (8) in acetic acid to produce the furan protected aminomaleimide 10b, followed by heating to remove furan through the retro Diels-Alder reaction. The structures of 2b, 3b, and 10b were established unequivocally by X-ray crystallography and other spectroscopic techniques. PMID- 15903347 TI - The first solid-phase synthesis of bis(oxazolinyl)pyridine ligands. AB - Chiral Pybox (pyridine-2,6-bis(oxazoline)) ligands can be cleanly and efficiently prepared on polystyrene support via a five-step solid-phase synthetic sequence. Cu(I)-complexed polymer-bound Pybox was used as a catalyst in the first heterogeneously catalyzed asymmetric addition of alkynes to imines. Best enantioselectivity was observed with (t)Bu-substituted oxazolines. PMID- 15903349 TI - Switching of prescription drugs to over-the-counter status: is it a good thing for the elderly? AB - Prescription medicines are increasingly being switched to over-the-counter (OTC, nonprescription) status in the developed world, with the support of government policy. These changes may provide greater choice for individuals and offer potential savings in government spending on health while expanding the market for pharmaceutical companies. However, there is concern regarding the safety of these reclassifications. Elderly people are the largest consumers of prescription and OTC medicines and are more vulnerable to drug adverse effects and the risks of multiple or inappropriate medications. Commonly purchased agents such as NSAIDs have recognised adverse effects which have been shown to be more common in the elderly. Furthermore, all sedatives, including antihistamines, have a propensity to cause falls in older people. As many doctors do not ask patients about OTC medicine use, problems related to use of these drugs may go undetected. Furthermore, the increased availability of OTCs may result in a delay in patients consulting medical practitioners for potentially serious conditions, although this has not so far been investigated. In the UK, the recent switch of a low-dose HMG-CoA reductase inhibitor (statin) to OTC status has caused concern. Although there might theoretically be some benefits from improved access to medications used in primary and secondary prevention of heart disease, the actual outcomes of use of this reduced dose of the statin will be difficult or impossible for patients or practitioners to monitor. OTC drug use implies a mutual responsibility for communication between patients and health professionals that in practice is not always achieved. Epidemiological research is needed to investigate patterns of OTC use and evaluate the potential risks of OTC medicines in elderly people. Governments, regulatory bodies, professionals and the drug industry have a responsibility to ensure that robust systems are in place if the increased use of OTC medicines by elderly people is to be safe and effective. PMID- 15903350 TI - Improving elder care by integrating geriatric expertise into medicare: a proposal. AB - The Medicare programme already funds a large proportion of the medical care for older Americans. However, geriatricians are an underutilized resource for improving the quality of care delivered with Medicare funding, particularly to the most complex patients. Recent legislative changes will increase the involvement of the Medicare programme in the area of outpatient medications. This offers an important opportunity to bring geriatric consultation to bear on a broad range of geriatric prescribing issues, including polypharmacy, pharmacological undertreatment and inappropriate prescription choice. PMID- 15903351 TI - Predicting and preventing adverse drug reactions in the very old. AB - The size of the elderly population has been increasing steadily for several years. Individuals in this age group often have several concomitant diseases that require treatment with multiple medications. These drugs, for various reasons and especially as a consequence of potential accumulation, may be associated with adverse reactions. Of the numerous factors that can favour the occurrence of these adverse drug reactions, the most important are the pathophysiological consequences of aging, particularly as these apply to the very old. Although absorption of drugs is not usually reduced in the elderly, diffusion, distribution and particularly elimination decline with age. Furthermore, while hepatic metabolic function is fairly normal, renal function is usually markedly depressed in very old individuals, and this can translate into clinical consequences if it is not taken into account. This is why, before administration of any drug in the elderly, evaluation of glomerular filtration rate is essential. Validated estimations such as those obtained from the classical Cockcroft-Gault formula or from more recent methodologies are required. In addition to reductions in various organ functions, factors connected with very old age such as frailty, falls, abnormal sensitivity to medications and polypathology, all of which tend to be more common in the last years of life, all directly impact on adverse drug reaction occurrence. Given these characteristics of the elderly population, the best way to reduce the prevalence of adverse drug reactions in this group is to limit drug prescription to essential medications, make sure that use of prescribed agents is clearly explained to the patient, give drugs for as short a period as possible, and periodically re-evaluate all use of drugs in the elderly. PMID- 15903352 TI - Combined lipoxygenase/cyclo-oxygenase inhibition in the elderly: the example of licofelone. AB - One of the categories of drugs most frequently used by the elderly, and probably the most commonly self-prescribed class of drug in this age group, is NSAIDs. However, NSAIDs are one of the primary causes of adverse drug reactions and are notorious for their gastric toxicity. They also inhibit renal function and reduce the efficacy of diuretics and ACE inhibitors, drugs that are commonly used by elderly patients. Recent studies have shown that cyclo-oxygenase (COX)-2 is important in renal physiology. This means that selective COX-2 inhibitors, while undoubtedly safer than NSAIDs in terms of gastric toxicity, are not devoid of renal toxicity (in addition to their now clearly established adverse effects on coronary heart disease risk). Both the gastric and renal toxicities induced by traditional NSAIDs and selective COX-2 inhibitors seem to be related to inhibition of prostaglandin, but not leukotriene, synthesis. Maintaining the correct balance between prostaglandins and leukotrienes is essential for continuing good health, but both classes of mediators also play an important role in the pathogenesis of several diseases.Recently, a new class of anti inflammatory drugs, the lipoxygenase (LOX)/COX inhibitors, has been developed as a means of simultaneously inhibiting the synthesis of prostaglandins, thromboxanes and leukotrienes. Inhibition of leukotriene synthesis increases anti inflammatory efficacy, particularly in rheumatic diseases, while reducing the risk of gastric damage. The LOX/COX inhibitor licofelone, which is currently in phase III trials, is the first of this new class and in the most advanced stage of development. Preliminary data with this drug seem promising, but further well designed clinical trials of this agent in the elderly will be necessary before a final evaluation is possible. PMID- 15903353 TI - Pharmacological management of severe postmenopausal osteoporosis. AB - The most devastating consequence of osteoporosis is bone fracture, particularly at the vertebral or femoral level. As defined by the WHO, patients with osteoporosis who have had one or more fragility fractures have severe osteoporosis. Those who sustain a vertebral fracture represent a particularly vulnerable group whose risk of another vertebral fracture within the following year is increased by a factor of 3-5. In addition, the presence of a vertebral fracture is associated with an increased risk of hip fracture. In light of these data, treatment of established osteoporosis is extremely important to prevent other fragility fractures. This review examines the therapies approved by the US FDA for the treatment of osteoporosis that have been shown to reduce the incidence of new fractures in patients with established osteoporosis. We evaluated the mechanisms of action, available formulations, efficacy in preventing fractures and increasing bone mineral density (BMD), duration of treatment, adverse effects and contraindications to use of alendronic acid (alendronate), risedronic acid (risedronate), calcitonin, raloxifene and teriparatide. All these drugs are able to prevent new vertebral fractures in patients with established osteoporosis. Only alendronic acid and risedronic acid have also been shown to reduce the risk of fracture at the femoral level, but they are contraindicated in patients with upper gastrointestinal diseases. Calcitonin is a good option in subjects with back pain because of its analgesic effect. Raloxifene is useful when patients have high plasma lipid levels or a family history of breast cancer. Teriparatide is indicated in subjects with very low BMD and multiple vertebral fractures. Patient characteristics should determine selection of therapy but the decision is always difficult and fraught with uncertainty. PMID- 15903354 TI - Dextropropoxyphene: safety and efficacy in older patients. AB - Dextropropoxyphene, alone or in combination with acetaminophen (paracetamol), is among the most frequently prescribed opioid analgesics in the elderly in the US despite the American Geriatric Society recommendation that its use should be restricted. However, this recommendation is based on expert opinion in an apparent absence of data. Accordingly, we conducted a literature search which identified nine studies that reported efficacy and safety data for dextropropoxyphene in predominantly older patients (> or = 55 years of age). These studies were evaluated to assess the efficacy and safety of dextropropoxyphene compared with other opioids and to evaluate whether safety and tolerability differed in older versus younger patients. The efficacy of dextropropoxyphene appeared to be similar to that of other analgesics, and its safety was comparable to that of other opioid analgesics. Although the adverse event profile suggests that elderly patients might have more frequent gastrointestinal and CNS complaints than younger patients treated with dextropropoxyphene, the frequency of reports appears similar to that of other opioids. The incidences of dizziness and somnolence were not significantly greater in older patients (1-2% and 0-21%, respectively) than in younger patients (8% and 13%, respectively). The absence of clinical studies directly addressing the safety and tolerability of dextropropoxyphene in elderly patients (>65 years of age) versus younger patients encumbers assessment of the validity of restricting its use in the elderly. Careful outcomes research is needed to assess the effectiveness and safety of dextropropoxyphene in older patients and to develop evidence-based risk/benefit prescribing criteria for use of this drug in this age group. PMID- 15903355 TI - The impact of age on rejection in kidney transplantation. AB - The time to failure of a renal allograft is determined by the initial function achieved after transplantation, the number and severity of insults to the graft, and a number of tissue characteristics. The insults a graft usually encounters include ischaemia/reperfusion injury, acute rejection episodes, drug-related nephrotoxicity, hypertension and hyperlipidaemia. Important tissue characteristics include susceptibility to injury and the ability of the tissue to repair damage. Elderly transplant recipients are considered poor immune responders but if a single acute rejection episode occurs this is more likely to significantly shorten graft and patient survival in this age group. Two issues have been identified with the use of old (>50 years of age) donor kidneys. First, compared with kidneys from younger donors, they have an increased incidence of acute interstitial rejection. Secondly, once a rejection episode occurs, the ability to mount a tissue repair process seems impaired. An explanation for the increased loss of grafts from old donors that have experienced acute rejection episodes is that such kidneys have fewer nephrons that function adequately and that the cumulated effect of damage results in an earlier demise of the graft compared with younger donor kidneys. Alternatively, graft parenchymal cells may undergo premature senescence or aging as a result of multiple injuries and repair. If progressive loss of renal mass or senescence is the mechanism responsible for increased graft loss, then it is expected that grafts from older donors will show a progressive decrease in function over time and that the rate of decline of function will correlate with donor age. We have suggested that increased graft loss of older donor kidneys results from increased incidence of acute rejection episodes in the early post-transplantation months together with a partly impaired ability to repair the tissue. Drug pharmacokinetic parameters are generally little influenced by age. However, the degree to which drugs suppress the immune system, and the extent to which kidneys from older donors are susceptible to the nephrotoxic effects of certain drugs, are unpredictable. There appears to be a more delicate balance between adequate immunosuppression and excess nonimmune toxicity in patients receiving older kidneys. Outcome parameters in elderly renal transplant recipients are currently dominated by increased death from infectious disease and drug-related (cardiovascular) causes. Increased susceptibility to nephrotoxic drugs, and to calcineurin inhibitors in particular, may be related to the increased risk of allograft failure experienced by the elderly as a surrogate for chronic allograft nephropathy. PMID- 15903357 TI - Evaluation of a standardized physical training program for basic combat training. AB - A control group (CG, n = 1,138) that implemented a traditional Basic Combat Training (BCT) physical training (PT) program was compared to an evaluation group (EG, n = 829) that implemented a PT program newly designed for BCT. The Army Physical Fitness Test (APFT) was taken at various points in the PT program, and injuries were obtained from a medical surveillance system. After 9 weeks of training, the proportion failing the APFT was lower in the EG than in the CG (1.7 vs. 3.3%, p = 0.03). After adjustment for initial fitness levels, age, and body mass index, the relative risk of an injury in the CG was 1.6 (95% confidence interval [CI] =1.2-2.0) and 1.5 (95% CI = 1.2-1.8) times higher than in the EG for men and women, respectively. The newly designed PT program resulted in higher fitness test pass rates and lower injury rates compared to a traditional BCT physical training program. PMID- 15903358 TI - Effects of basic training in the British Army on regular and reserve army personnel. AB - The aim of this study was to compare changes in aerobic fitness and body composition in response to British Army (regular) and Territorial Army (reserve) basic training. Eleven regular recruits, 14 reserve recruits, and 20 controls completed the study (all males). Initially, reserve recruits were significantly older and heavier and had greater fat-free mass (FFM; 64.6 vs. 59.3 kg) and lower maximal oxygen uptake (Vo(2)max; 39.1 vs. 43.9 ml.kg(-1).min(-1)) than regular recruits. Both regular and reserve training significantly increased FFM and Vo(2)max and decreased percentage body fat. Regular training produced a greater increase in Vo(2)max than reserve training (13.1 vs. 7.6%, p < 0.0005). Reserve training produced a greater increase in body mass (2.2 vs. 0.9 kg, p = 0.019) and tended to produce a greater increase in FFM (2.6 vs. 1.6 kg, p = 0.062). Although both training programs improve aerobic fitness and body composition, increasing the volume of physical training in the reserve training program would probably enhance the training adaptations achieved. PMID- 15903359 TI - Effect of low-dose, short-duration creatine supplementation on anaerobic exercise performance. AB - To examine the efficacy of a low-dose, short-duration creatine monohydrate supplement, 40 physically active men were randomly assigned to either a placebo or creatine supplementation group (6 g of creatine monohydrate per day). Testing occurred before and at the end of 6 days of supplementation. During each testing session, subjects performed three 15-second Wingate anaerobic power tests. No significant (p > 0.05) group or time differences were observed in body mass, peak power, mean power, or total work. In addition, no significant (p > 0.05) differences were observed in peak power, mean power, or total work. However, the change in the rate of fatigue of total work was significantly (p < 0.05) lower in the creatine supplementation group than in the placebo group, indicating a reduced fatigue rate in subjects supplementing with creatine compared with the placebo. Although the results of this study demonstrated reduced fatigue rates in patients during high-intensity sprint intervals, further research is necessary in examining the efficacy of low-dose, short-term creatine supplementation. PMID- 15903360 TI - Effects of acute creatine loading with or without carbohydrate on repeated bouts of maximal swimming in high-performance swimmers. AB - The addition of carbohydrate (CHO) to an acute creatine (Cr) loading regimen has been shown to increase muscle total creatine content significantly beyond that achieved through creatine loading alone. However, the potential ergogenic effects of combined Cr and CHO loading have not been assessed. The purpose of this study was to compare swimming performance, assessed as mean swimming velocity over repeated maximal intervals, in high-performance swimmers before and after an acute loading regimen of either creatine alone (Cr) or combined creatine and carbohydrate (Cr + CHO). Ten swimmers (mean +/- SD of age and body mass: 17.8 +/- 1.8 years and 72.3 +/- 6.8 kg, respectively) of international caliber were recruited and were randomized to 1 of 2 groups. Each swimmer ingested five 5 g doses of creatine for 4 days, with the Cr + CHO group also ingesting approximately 100 g of simple CHO 30 minutes after each dose of creatine. Performance was measured on 5 separate occasions: twice at "baseline" (prior to intervention, to assess the repeatability of the performance test), within 48 hours after intervention, and then 2 and 4 weeks later. All subjects swam faster after either dietary loading regimen (p < 0.01, both regimens); however, there was no difference in the extent of improvement of performance between groups. In addition, all swimmers continued to produce faster swim times for up to 4 weeks after intervention. Our findings suggest that no performance advantage was gained from the addition of carbohydrate to a creatine-loading regimen in these high caliber swimmers. PMID- 15903361 TI - Myosin heavy chain isoform expression and in vivo isometric performance: a regression model. AB - This investigation estimated the amount of variance in voluntary in vivo muscle performance that can be explained by relative myosin heavy chain (MHC) isoform expression. The role of the relative expression of these proteins in relation to in vitro force and velocity performance is well understood, but the in vivo model is less clear. Twenty-two men and women (mean +/- SD age, 27 +/- 6 years) performed isometric knee extensor actions in which peak force and rate of force development (RFD) were measured. The results of regression analysis showed that the inclusion of MHC IIb explained a significant (19.9%, p < 0.05) amount of variance in relative peak force (adjusted for muscle mass) and 14.1% of the variance in the first half of the rise phase of the force-time curve (RFD(0-50%)) (p < 0.1). The addition of MHC I into this model explained a significant (p < 0.05) amount of variance above that accounted for by MHC IIb in RFD (45.4%), RFD(0-50%) (50.8%), and RFD(50-100%) (second half of the rise phase of the force time curve) (37.4%). Since the percentage of MHC IIb is reduced rather quickly with training, these data suggest that peak force may also be affected quickly by training. The percentage of MHC I has a longer course for change with training; therefore, it may be inferred that the greatest changes in RFD variables will likely occur during a longer course. PMID- 15903362 TI - Effects of acute aerobic and anaerobic exercise on blood markers of oxidative stress. AB - The purpose of this study was to compare oxidative modification of blood proteins, lipids, DNA, and glutathione in the 24 hours following aerobic and anaerobic exercise using similar muscle groups. Ten cross-trained men (24.3 +/- 3.8 years, [mean +/- SEM]) performed in random order 30 minutes of continuous cycling at 70% of Vo(2)max and intermittent dumbbell squatting at 70% of 1 repetition maximum (1RM), separated by 1-2 weeks, in a crossover design. Blood samples taken before, and immediately, 1, 6, and 24 hours postexercise were analyzed for plasma protein carbonyls (PC), plasma malondialdehyde (MDA), and whole-blood total (TGSH), oxidized (GSSG), and reduced (GSH) glutathione. Blood samples taken before and 24 hours postexercise were analyzed for serum 8-hydroxy 2'-deoxyguanosine (8-OHdG). PC values were greater at 6 and 24 hours postexercise compared with pre-exercise for squatting, with greater PC values at 24 hours postexercise for squatting compared with cycling (0.634 +/- 0.053 vs. 0.359 +/- 0.018 nM.mg protein(-1)). There was no significant interaction or main effects for MDA or 8-OHdG. GSSG experienced a short-lived increase and GSH a transient decrease immediately following both exercise modes. These data suggest that 30 minutes of aerobic and anaerobic exercise performed by young, cross-trained men (a) can increase certain biomarkers of oxidative stress in blood, (b) differentially affect oxidative stress biomarkers, and (c) result in a different magnitude of oxidation based on the macromolecule studied. Practical applications: While protein and glutathione oxidation was increased following acute exercise as performed in this study, future research may investigate methods of reducing macromolecule oxidation, possibly through the use of antioxidant therapy. PMID- 15903363 TI - Preferential vastus medialis oblique activation achieved as a treatment for knee disorders. AB - The purpose of this study was to compare the effect of an open-stance cycling protocol (OSCP) with the traditional cycling foot position (TCFP) for preferential vastus medialis oblique (VMO) muscle activation, measured by surface electromyography (SEMG), and preferential VMO activation as defined by achieving significantly increased VMO/VL (vastus lateralis muscle) ratio values. Forty subjects of both sexes participated, 18 symptomatic with patellofemoral pain and 22 control subjects; ages ranged from 18 to 60 years (mean = 28.7 +/- 8 years). The OSCP and TCFP were ridden in randomized order while SEMG recordings were taken of the VMO and VL muscles, collecting the mean of peak amplitudes to calculate VMO/VL ratio values. The SEMG readings were taken 4 times per testing session with randomized resistance and a consistent cycling cadence of 85 rpm. The OSCP displayed preferential VMO activation for all subject groups (F = 40.47, p = 0.0001), and this study revealed a protocol that effectively treats patellofemoral pain. PMID- 15903364 TI - Influence of moderate prophylactic compression on sport performance. AB - This study examined the impact of using elasticized compression shorts on performance measures and proprioception at the hip. Thirteen healthy subjects completed 2 randomized testing sessions-one while wearing the Coreshorts compression shorts and one while not wearing the shorts. During each trial, active range of motion at the hip; joint angle replication during hip flexion, abduction, and hyperextension; leg power; agility; speed; and aerobic endurance were measured, and subjective information pertaining to the fit of the shorts was collected. The use of the prophylactic brace did not limit performance on any measure except active range of motion during hip flexion (p < 0.05). Subjective data revealed 93.3% of subjects felt the shorts were supportive, although proper fit was an issue. The present results support the use of moderate compression at and around the hip for the purpose of injury prevention. Continued research is necessary to determine the efficacy of hip bracing within an injured population and their potential prophylactic benefit for active individuals. PMID- 15903365 TI - Kinematics of biophysically asymmetric limbs within rate of velocity development. AB - The purpose of this study was to investigate the movement speed characteristics of 2 intrinsically different limbs. Twenty subjects volunteered to participate (10 men and 10 women). Each subject performed 5 repetitions of concentric knee and elbow extension and flexion movements at 60 through 500 d.s(-1) on an isokinetic dynamometer. Kinematic data were collected at 1,000 Hz and separated into rate of velocity development (RVD) and peak torque. Results demonstrated a significant (p < 0.05) main effect for sex for RVD and peak torque. Significant (p < 0.05) differences were also demonstrated between knee and elbow RVD and between knee and elbow peak torque at every speed tested. Neither knee and elbow RVD nor peak torque demonstrated any significant Pearson correlations at any speed tested (r = -0.17-0.41). These results collectively point to the specificity of limb speed and torque as a result of biophysical differences such as length and mass. Therefore, strength and speed may be modulated by neuromotor patterns that differ based on individual limbs. PMID- 15903366 TI - Activation of vastus medialis obliquus among individuals with patellofemoral pain syndrome. AB - The purpose of this study was to determine whether various positions of the lower extremity affect the muscle activity of the vastus medialis obliquus (VMO) differently during both open and closed kinetic chain exercise conditions among patients with patellofemoral pain syndrome (PFPS). Patients who presented with symptoms consistent with PFPS completed a series of open kinetic chain and closed kinetic chain exercises in which VMO activity was measured and compared. Statistical analysis revealed that there is less than a 0.001% (open kinetic chain) or 0.005% (closed kinetic chain) chance that all positions activate the VMO equally. In open kinetic exercise, maximum VMO activity was achieved with terminal knee extension with medial tibial rotation. During closed kinetic exercises, squats with external rotation were preferred for maximum VMO activation. Therefore, our results highlight the importance of including both the open and closed kinetic chain exercises into rehabilitation programs for patients with PFPS. PMID- 15903367 TI - Evaluation of physiological responses during recovery following three resistance exercise programs. AB - The present study was conducted to examine (a) whether there is an association between maximal oxygen uptake (Vo(2)max) and reduction in postexercise heart rate (HR) and blood lactate concentrations ([La]) following resistance exercise and (b) how intensity and Volume of resistance exercise affect postexercise Vo(2). Eleven regularly weight-trained males (20.8 +/- 1.3 years; 96.2 +/- 14.4 kg, 182.4 +/- 7.3 cm) underwent 4 sets of squat exercise on 3 separate occasions that differed in both exercise intensity and volume. During each testing session, subjects performed either 15 repetitions.set(-1) at 60% of 1 repetition maximum (1RM) (L), 10 repetitions.set(-1) at 75% of 1RM (M), or 4 repetitions.set(-1) at 90% of 1RM (H). During each exercise, Vo(2) and HR were measured before (PRE), immediately post (IP), and at 10 (10P), 20 (20P) 30 (30P), and 40 (40P) minutes postexercise. The [La] was measured at PRE, IP, 20P, and 40P. Decrease in HR (DeltaHR) was determined by subtracting HR at 10P from that at IP, whereas decrease in [La] (Delta[La]) was computed by subtracting [La] at 20P from that at IP. A significant correlation (p < 0.05) was found between Vo(2)max and DeltaHR in all exercise conditions. A significant correlation (p < 0.05) was also found between Vo(2)max and Delta[La] in L and M but not in H. The Vo(2) was higher (p < 0.05) during M than H at IP and 10P, while no difference was seen between L and M and between L and H. These results indicate that those with greater aerobic capacity tend to have a greater reduction in HR and [La] during recovery from resistance exercise. In addition, an exercise routine performed at low to moderate intensity coupled with a moderate to high exercise volume is most effective in maximizing caloric expenditure following resistance exercise. PMID- 15903368 TI - Physical fitness of an industrial fire department vs. a municipal fire department. AB - Both industrial and municipal firefighters need to maintain high levels of physical fitness and minimize cardiovascular risk factors. The nature of firefighter responsibilities in industrial and municipal settings may vary, affecting the ability to sustain high levels of physical fitness. We compared the working conditions, physical fitness, and exercise training practices of an industrial fire department (n = 17) to those of a nearby municipal fire department (n = 55). After informed consent, aerobic capacity, muscular strength, muscular endurance, body composition, flexibility, blood lipid concentrations, and blood pressure levels were measured. Exercise training practices and related factors were assessed using a questionnaire. Despite programmatic differences, these departments demonstrated similar, relatively high degrees of physical fitness and similar blood lipid concentrations, blood pressure levels, and cardiac risk factors. It is recommended that fire departments involve appropriately trained staff, schedule on-duty times for exercise, offer well equipped exercise facilities, and follow National Strength and Conditioning Association (NSCA) and American College of Sports Medicine (ACSM) guidelines for exercise conditioning in order to maintain a high degree of physical fitness. PMID- 15903369 TI - Measurement of talent in team handball: the questionable use of motor and physical tests. AB - Testing for selection is one of the most important fundamentals in any multistep sport program. In most ball games, coaches assess motor, physical, and technical skills on a regular basis in early stages of talent identification and development. However, selection processes are complex, are often unstructured, and lack clear-cut theory-based knowledge. For example, little is known about the relevance of the testing process to the final selection of the young prospects. The purpose of this study was to identify motor, physical, and skill variables that could provide coaches with relevant information in the selection process of young team handball players. In total, 405 players (12-13 years of age at the beginning of the testing period) were recommended by their coaches to undergo a battery of tests prior to selection to the Junior National Team. This number is the sum of all players participating in the different phases of the program. However, not all of them took part in each testing phase. The battery included physical measurements (height and weight), a 4 x 10-m running test, explosive power tests (medicine ball throw and standing long jump), speed tests (a 20-m sprint from a standing position and a 20-m sprint with a flying start), and a slalom dribbling test. Comparisons between those players eventually selected to the Junior National Team 2-3 years later with those not selected demonstrated that only the skill test served as a good indicator. In all other measurements, a wide overlap could be seen between the results of the selected and nonselected players. It is suggested that future studies investigate the usefulness of tests reflecting more specific physical ability and cognitive characteristics. PMID- 15903370 TI - Relationship between hockey skating speed and selected performance measures. AB - The objective of this study was to determine the relationship between specific performance measures and hockey skating speed. Thirty competitive secondary school and junior hockey players were timed for skating speed. Off-ice measures included a 40-yd (36.9-m) sprint, concentric squat jump, drop jump, 1 repetition maximum leg press, flexibility, and balance ratio (wobble board test). Pearson product moment correlations were used to quantify the relationships between the variables. Electromyographic (EMG) activity of the dominant vastus lateralis and biceps femoris was monitored in 12 of the players while skating, stopping, turning, and performing a change-of-direction drill. Significant correlations (p < 0.005) were found between skating performance and the sprint and balance tests. Further analysis demonstrated significant correlations between balance and players under the age of 19 years (r = -0.65) but not those over 19 years old (r = -0.28). The significant correlations with balance suggested that stability may be associated with skating speed in younger players. The low correlations with drop jumps suggested that short contact time stretch-shortening activities (i.e., low amplitude plyometrics) may not be an important factor. Electromyographic activities illustrated the very high activation levels associated with maximum skating speed. PMID- 15903371 TI - Aerobic and resistance exercise sequence affects excess postexercise oxygen consumption. AB - Excess postexercise oxygen consumption (EPOC) may describe the impact of previous exercise on energy metabolism. Ten males completed Resistance Only, Run Only, Resistance-Run, and Run-Resistance experimental conditions. Resistance exercise consisted of 7 lifts. Running consisted of 25 minutes of treadmill exercise. Vo(2) was determined during treadmill exercise and after each exercise treatment. Our findings indicated that treadmill exercise Vo(2) was significantly higher for Resistance-Run compared with Run-Resistance and Resistance Only at all time intervals. At 10 minutes postexercise, Vo(2) was greater for Resistance Only and Run-Resistance than for Resistance-Run. At 20 and 30 minutes, Vo(2) following Resistance Only was significantly greater than following Run Only. In conclusion, EPOC is greatest following Run-Resistance; however, treadmill exercise is more physiologically difficult following resistance exercise. Furthermore, the sequence of resistance and treadmill exercise influences EPOC, primarily because of the effects of resistance exercise rather than the exercise combination. We recommend performing aerobic exercise before resistance exercise when combining them into 1 exercise session. PMID- 15903372 TI - Acute muscle stretching inhibits muscle strength endurance performance. AB - Since strength and muscular strength endurance are linked, it is possible that the inhibitory influence that prior stretching has on strength can also extend to the reduction of muscle strength endurance. To date, however, studies measuring muscle strength endurance poststretching have been criticized because of problems with their reliability. The purpose of this study was twofold: both the muscle strength endurance performance after acute static stretching exercises and the repeatability of those differences were measured. Two separate experiments were conducted. In experiment 1, the knee-flexion muscle strength endurance exercise was measured by exercise performed at 60 and 40% of body weight following either a no-stretching or stretching regimen. In experiment 2, using a test-retest protocol, a knee-flexion muscle strength endurance exercise was performed at 50% body weight on 4 different days, with 2 tests following a no-stretching regimen (RNS) and 2 tests following a stretching regimen (RST). For experiment 1, when exercise was performed at 60% of body weight, stretching significantly (p < 0.05) reduced muscle strength endurance by 24%, and at 40% of body weight, it was reduced by 9%. For experiment 2, reliability was high (RNS, intraclass correlation = 0.94; RST, intraclass correlation = 0.97). Stretching also significantly (p < 0.05) reduced muscle strength endurance by 28%. Therefore, it is recommended that heavy static stretching exercises of a muscle group be avoided prior to any performances requiring maximal muscle strength endurance. PMID- 15903373 TI - Reliability of the Lode Excalibur Sport Ergometer and applicability to Computrainer electromagnetically braked cycling training device. AB - New technology allows cyclists to train via power output (PO) in addition to heart rate (HR). For those athletes undertaking seasonal laboratory testing (e.g., Vo(2), lactate threshold), it is imperative that athletes be able to directly apply this information to their training device. We examined the reliability of a standardized laboratory ergometer (Lode Excalibur Sport) and its applicability to an electromagnetically braked ergometer (Computrainer) in 2 phases. Phase I (n = 12) examined the reliability of the Lode. Phase II (n = 14) compared the Lode to the Computrainer using a randomized, counterbalance assignment. Following warm-up, each trial started at 100 W, progressing 50 W every 3 minutes to exhaustion. Outcomes were time-to-exhaustion (TTE), peak PO (W) (PO(peak)), peak HR (HR(peak)), and ventilatory (VT) and respiratory compensation (RCP) thresholds. We used a repeated measures analysis of variance (ANOVA), Tukey post hoc analysis, regression analysis, Bland-Altman plots, and coefficient of variation (CV) analysis for each variable. During phase I, we found no significant difference for any variable, minimal dispersion of Vo(2) during Bland-Altman analysis, and a low CV at each test stage ( 0.05) across torque levels. For the isokinetic muscle actions, EMG amplitude (R(2) = 0.988) and MMG amplitude (R(2) = 0.933) increased to PT, but there were no significant mean changes with torque for EMG MPF or MMG MPF. The different torque-related responses for EMG and MMG amplitude and MPF may reflect differences in the motor control strategies that modulate torque production for isometric vs. dynamic muscle actions. These results support the findings of others and suggest that isometric torque production was modulated by a combination of recruitment and firing rate, whereas dynamic torque production was modulated primarily through recruitment. PMID- 15903385 TI - Effect of potentiation and stretching on maximal force, rate of force development, and range of motion. AB - The purpose of this investigation was to compare the effects of stretching vs. potentiation on subsequent maximal force and rate of force development capabilities of subjects in an isometric squat. Ten male collegiate athletes participated as subjects in this study. Subjects were tested during 3 separate sessions that involved joint range of motion (ROM) measurements of the lower body and isometric squat trials on a force plate to determine peak force (PF) and rate of force development (RFD) values. One testing session was preceded by 10 minutes of quiet sitting (C), 1 by a 30-minute lower-body stretching protocol (S), and 1 by 3 sets of a leg press exercise using 90% of the subjects' previously determined 1 repetition maximum (P). Three repetitions were performed for each set of the leg press, with a 3-minute rest period between each set. PF during the isometric squat was not significantly different following any of the 3 conditions (C: 100.0 +/- 0.0%, S: 101.2 +/- 6.5%, P: 98.6 +/- 6.2%). However, RFD was significantly lower in P (87.5 +/- 12.8%) compared with both C (100.0 +/- 0.0%) and S (102.6 +/- 18.5%). Significant improvement in ROM occurred only following P. It appears the potentiation protocol used in the current investigation may actually have had fatiguing effects instead of potentiating effects, but it did result in significant increases in ROM. PMID- 15903386 TI - The acute effects of prior dynamic resistance exercise using different loads on subsequent upper-body explosive performance in resistance-trained men. AB - The purpose of this study was to determine if explosive upper-body performance could be improved when it was preceded by conditioning contraction protocols that incorporate resistance exercise. Providing that performance was enhanced, it was also the intention to determine the optimal conditioning contraction load for enhancing performance. Eight recreationally trained men completed 4 experimental sessions. Each session consisted of a warm-up, 3 bench press throws (pre), a conditioning protocol, and 3 bench press throws (post). The different conditioning protocols consisted of 5 bench press repetitions using 100, 75, or 50% of 5 repetition maximum (5RM) strength. The fourth protocol, in which no repetitions were completed, acted as a control. Participants performed each conditioning protocol on a different day, and the order in which the protocols were performed was randomized. Average power, assessed during the bench press throws, was determined for the starting segment and the end segment (point of bar release) for each throw. Comparisons in average power, for each segment of the bench press 1RM, were made between the pre- and postconditioning protocol bench press throws. None of the conditioning protocols had an effect on bench press throw performance in either of the 2 segments of the movement. The results suggest there is no performance advantage when explosive upper-body movement is preceded by resistance exercise of varying loads. Alternatively, the performance of a set of resistance exercise did not compromise explosive upper-body performance. Considering this, training methods that combine both resistance exercise and plyometric-like exercise may offer a practical and time-efficient training system. PMID- 15903387 TI - Short-term effects on lower-body functional power development: weightlifting vs. vertical jump training programs. AB - Among sport conditioning coaches, there is considerable discussion regarding the efficiency of training methods that improve lower-body power. Heavy resistance training combined with vertical jump (VJ) training is a well-established training method; however, there is a lack of information about its combination with Olympic weightlifting (WL) exercises. Therefore, the purpose of this study was to compare the short-term effects of heavy resistance training combined with either the VJ or WL program. Thirty-two young men were assigned to 3 groups: WL = 12, VJ = 12, and control = 8. These 32 men participated in an 8-week training study. The WL training program consisted of 3 x 6RM high pull, 4 x 4RM power clean, and 4 x 4RM clean and jerk. The VJ training program consisted of 6 x 4 double-leg hurdle hops, 4 x 4 alternated single-leg hurdle hops, 4 x 4 single-leg hurdle hops, and 4 x 4 40-cm drop jumps. Additionally, both groups performed 4 x 6RM half-squat exercises. Training volume was increased after 4 weeks. Pretesting and posttesting consisted of squat jump (SJ) and countermovement jump (CMJ) tests, 10 and 30-m sprint speeds, an agility test, a half-squat 1RM, and a clean-and-jerk 1RM (only for WL). The WL program significantly increased the 10-m sprint speed (p < 0.05). Both groups, WL and VJ, increased CMJ (p < 0.05), but groups using the WL program increased more than those using the VJ program. On the other hand, the group using the VJ program increased its 1RM half-squat strength more than the WL group (47.8 and 43.7%, respectively). Only the WL group improved in the SJ (9.5%). There were no significant changes in the control group. In conclusion, Olympic WL exercises seemed to produce broader performance improvements than VJ exercises in physically active subjects. PMID- 15903388 TI - Percutaneous electrical stimulation in strength training: an update. AB - Numerous studies have used percutaneous electrical stimulation (PES) in the context of training programs to develop strength and physical performance in healthy populations (sedentary or trained). Significant increases in muscle and fiber cross-sectional area, isokinetic peak torque, maximal isometric and dynamic strength, and motor performance skills have been found after PES training. These strength gains are explained on the basis of the characteristics of PES motor units (MUs) recruitment: (a) a continuous and exhausting contractile activity in the same pool of MUs during the entire exercise period, (b) a supramaximal temporal recruitment imposed by the high frequency chosen (up to 40 Hz), and (c) a synchronous recruitment of neighboring fibers. The PES training method is complementary to voluntary training, mainly because the application of PES causes an unconventional spatial recruitment of MUs that, depending on the muscular topography, may entail the preferential recruitment of the fast-twitch MUs. In addition, the method does not specifically develop elasticity in skeletal muscle, and it must be accompanied by a technical workout. PMID- 15903389 TI - Electromyographic activity of the pectoralis major and anterior deltoid muscles during three upper-body lifts. AB - The purpose of this study was to examine the differences in activation levels and times of activation for the pectoralis major and anterior deltoid when performing the concentric phase of 3 upper-body lifts. Twelve college-age men and women with various degrees of lifting experience performed 3 repetitions using the 6 repetition maximum in a barbell bench press, dumbbell bench press, and dumbbell fly while being monitored for electromyographic activity in both muscles. Motor unit activation of both muscles was not significantly different during all 3 lifts. However, dumbbell flys had significantly less relative time of activation than did barbell or dumbbell bench presses. Therefore, dumbbell flys may be better suited as an auxiliary lift, whereas barbell and dumbbell bench presses may be used interchangeably in training programs. The compatibility of the barbell and dumbbell bench presses may aid lifters in overcoming training plateaus by alternating exercises for the same muscle groups. PMID- 15903390 TI - Postactivation potentiation and its practical applicability: a brief review. AB - It has been suggested that postactivation potentiation (PAP) may be manipulated to enhance both acute performance and chronic adaptation. PAP refers to the phenomenon by which acute muscle force output is enhanced as a result of contractile history. Evidence exists regarding the existence of PAP. However, the determination of methods to best manipulate and exploit PAP remains elusive. Studies to date would seem to indicate that the practical applicability of PAP in terms of enhancing athletic performance is limited. PMID- 15903391 TI - Vibration training: an overview of the area, training consequences, and future considerations. AB - The effects of vibration on the human body have been documented for many years. Recently, the use of vibration for improving the training regimes of athletes has been investigated. Vibration has been used during strength-training movements such as elbow flexion, and vibration has also been applied to the entire body by having subjects stand on vibration platforms. Exposure to whole-body vibration has also resulted in a significant improvement in power output in the postvibratory period and has been demonstrated to induce significant changes in the resting hormonal profiles of men. In addition to the potential training effects of vibration, the improvement in power output that is observed in the postvibratory period may also lead to better warm-up protocols for athletes competing in sporting events that require high amounts of power output. These observations provide the possibility of new and improved methods of augmenting the training and performance of athletes through the use vibration training. Despite the potential benefits of vibration training, there is substantial evidence regarding the negative effects of vibration on the human body. In conclusion, the potential of vibration treatment to enhance the training regimes of athletes appears quite promising. It is essential though that a thorough understanding of the implications of this type of treatment be acquired prior to its use in athletic situations. Future research should be done with the aim of understanding the biological effects of vibration on muscle performance and also the effects of different vibration protocols on muscle performance. PMID- 15903392 TI - Normalizing physical performance tests for body size: a proposal for standardization. AB - There is a lack of standardized methodology for normalizing various indices of muscle strength and movement performance tests for differences in body size in human movement-related disciplines. Most of the data presented in the literature have been body size dependent, which precludes both comparisons between subjects and establishment of standards for specific subject populations. The goal of the present review was to propose standardized tests that normalize physical performance tests to body size. Specifically, we propose (a) using an allometric normalization based on theoretical models that presume geometric similarity, (b) using classification of performance tests based on particular values of the allometric parameters required for normalization, and (c) using a simple "performance index" that represents an individual or group performance relative to a reference population. Correspondences between theory and experimental findings and limitations are discussed. PMID- 15903393 TI - A cetylated fatty acid topical cream with menthol reduces pain and improves functional performance in individuals with arthritis. AB - This investigation was an extension of a previous study conducted in our laboratory in which we showed that 1 month of treatment with a topical cream (Celadrin) consisting of cetylated fatty acids was effective for reducing pain and improving functional performance in individuals with osteoarthritis (OA) of the knee (Kraemer et al., Journal of Rheumatology, 2004). We wanted to verify that the addition of menthol to the compound would produce a similar percentage of improvement in therapeutic effects. We used a single treatment group with a pre-post experimental design to examine % treatment changes. Individuals diagnosed with OA of the knee (N = 10; age, 66.4 +/- 11.5 years) and severe pain (e.g., OA, rheumatoid arthritis) of the elbow (N = 8; age, 59.1 +/- 18.2 years) and wrist (N = 10; age, 60.3 +/- 16.8 years) were tested for pain and functional performance before and after 1 week of treatment with a topical cream consisting of cetylated fatty acids and menthol applied twice per day. In individuals with knee OA, significant improvements in stair-climbing ability (about 12%), "up-and go" performance (about 12%), balance and strength (about 16.5%), and range of motion (about 3.5%) were observed, as were reductions in pain. In individuals with severe pain of the elbow and wrist, significant improvements in dynamic (about 22 and 24.5%, respectively) and isometric (about 33 and 42%, respectively) local muscular endurance were observed, as was a reduction in pain. Neither group demonstrated significant changes in maximal grip strength or maximal force production. One week of treatment with a topical cream consisting of cetylated fatty acids and menthol was similarly effective for reducing pain and improving functional performance in individuals with arthritis of the knee, elbow, and wrist. The % changes were consistent with our prior work on the compound without menthol. Further work is needed to determine the impact of menthol in such a cream. Nevertheless, our data support the use of a topical cream consisting of cetylated fatty acids (with or without menthol) for enhancing the potential for exercise training in this population. PMID- 15903395 TI - Universal power law tails of time correlation functions. AB - The universal power law tails of single particle and multiparticle time correlation functions are derived from a unifying point of view, solely using the hydrodynamic modes of the system. The theory applies to general correlation functions and to systems more general than classical fluids. Moreover, it is argued that the collisional transfer part of the stress-stress correlation function in dense classical fluids has the same long-time tail approximately t(-1 d/2) as the velocity autocorrelation function in Lorentz gases. PMID- 15903396 TI - Finite-energy extension of a lattice glass model. AB - We extend a previously studied lattice model of particles with infinite repulsions to the case of finite-energy interactions. The phase diagram is studied using grand canonical Monte Carlo simulation. Simulations of dynamical phenomena are made using the canonical ensemble. We find interesting order disorder transitions in the equilibrium phase diagram and identify several anomalous regimes of diffusivity. These phenomena may be relevant to the case of strong orientational bonding near freezing. PMID- 15903397 TI - Stationary state volume fluctuations in a granular medium. AB - A statistical description of static granular material requires ergodic sampling of the phase space spanned by the different configurations of the particles. We periodically fluidize a column of glass beads and find that the sequence of volume fractions phi of postfluidized states is history independent and Gaussian distributed about a stationary state. The standard deviation of phi exhibits, as a function of phi, a minimum corresponding to a maximum in the number of statistically independent regions. Measurements of the fluctuations enable us to determine the compactivity X , a temperaturelike state variable introduced in the statistical theory of Edwards and Oakeshott [Physica A 157, 1080 (1989)]. PMID- 15903398 TI - Convection-driven pattern formation in phase-separating binary fluids. AB - Using a thermal-lattice Boltzmann model, we examine the rich phase behavior that develops when partially miscible fluids evolve in the presence of a vertical temperature gradient, which encompasses the critical temperature T(c) of the mixture. In particular, a binary AB fluid is confined between two plates in a gravitational field. The upper plate is fixed below T(c) and hence, the nearby fluid phase separates into A-rich and B-rich domains. The lower plate is fixed above the temperature T(c), and the surrounding fluid is in the homogeneous phase. A coupling between convection (driven by the temperature gradient) and phase separation gives rise to unique pattern formation. A number of regimes are identified: regularly spaced stripes, convective steady-state columns, the periodic disturbance of these columns, and finally, chaotic dripping from the upper surface. These results highlight dynamical behavior in partially miscible mixtures. PMID- 15903399 TI - Anomalous glassy relaxation near the isotropic-nematic phase transition. AB - Dynamical heterogeneity in a system of Gay-Berne ellipsoids near its isotropic nematic (I-N) transition, and also in an equimolar mixture of Lennard-Jones spheres and Gay-Berne ellipsoids in deeply supercooled regime, is probed by the time evolution of non-Gaussian parameters (NGP). The appearance of a dominant second peak in the rotational NGP near the I-N transition signals the growth of pseudonematic domains. Surprisingly, such a second peak is instead observed in the translational NGP for the glassy binary mixture. Localization of orientational motion near the I-N transition is found to be responsible for the observed anomalous orientational relaxation. PMID- 15903400 TI - Free-energy formalism for particle adsorption. AB - The equilibrium properties of particle adsorption is investigated theoretically. The model relies on a free-energy formulation which allows us to generalize the Maxwell-Boltzmann description to solutions for which the bulk volume fraction of potentially adsorbed particles is very high. As an application we consider the equilibrium physical adsorption of neutral and charged particles from solution onto two parallel adsorbing surfaces. PMID- 15903401 TI - Deterministic directed transport of inertial particles in a flashing ratchet potential. AB - Deterministic directed transport of inertia particles in a periodically on-off ratchet potential is investigated. We find that the directed transport can be induced by a finite inertia; i.e., in the overdamped case, no directed motion of the system can be observed. It is shown that a critical threshold of the ratchet asymmetry is required for the system to achieve a net current. Directed transport can be greatly enhanced when the coupling strength of particles is increased. An appropriate match of the coupling, the flashing period, and the damping can give rise to the best efficiency of transport. The commensurate effect on the directed transport, which originates from the spatial competition between the period of the potential and the static length of the coupling, is analyzed. PMID- 15903402 TI - Equilibrium statistics of a slave estimator in Langevin processes. AB - We analyze the statistics of an estimator, denoted by xi(t) and referred to as the slave, for the equilibrium susceptibility of a one dimensional Langevin process x(t) in a potential phi (x). The susceptibility can be measured by evolving the slave equation in conjunction with the original Langevin process. This procedure yields a direct estimate of the susceptibility and avoids the need, when performing numerical simulations, to include applied external fields explicitly. The success of the method, however, depends on the statistical properties of the slave estimator. The joint probability density function for x(t) and xi(t) is analyzed. In the case where the potential of the system has a concave component the probability density function of the slave acquires a power law tail characterized by a temperature dependent exponent. Thus we show that while the average value of the slave, in the equilibrium state, is always finite and given by the fluctuation-dissipation relation, higher moments and indeed the variance may show divergences. The behavior of the power law exponent is analyzed in a general context and it is calculated explicitly in some specific examples. Our results are confirmed by numerical simulations and we discuss possible measurement discrepancies in the fluctuation dissipation relation which could arise due to this behavior. PMID- 15903403 TI - Order-disorder phase transition in random-walk networks. AB - In this paper we study in detail the behavior of random-walk networks (RWN's). These networks are a generalization of the well-known random Boolean networks (RBN's), a classical approach to the study of the genome. RWN's are also discrete networks, but their response is defined by small variations in the state of each gene, thus being a more realistic representation of the genome and a natural bridge between discrete and continuous models. RWN's show a clear transition between order and disorder. Here we explicitly deduce the formula of the critical line for the annealed model and compute numerically the transition points for quenched and annealed models. We show that RBN's and the annealed model of RWN's act as an upper and a lower limit for the quenched model of RWN's. Finally we calculate the limit of the annealed model for the continuous case. PMID- 15903404 TI - Asymmetric probability densities in symmetrically modulated bistable devices. AB - A Brownian particle hopping in a symmetric double-well potential can be statistically confined into a single well by the simultaneous action of (a) two periodic input signals, one tilting the minima and the other one modulating the barrier height, and (b) an additive and a purely multiplicative random signal, generated by a unique source and thus preserving a certain degree of statistical correlation. The underlying gating mechanism is quite robust when compared, for instance, with biharmonic rocking. In view of technological implementation, asymmetric confinement through gating can be conveniently maximized by tuning the input signal parameters (correlation time, phase-time lag, amplitudes), thus revealing a resonant localization mechanism of general applicability. PMID- 15903405 TI - Delay Fokker-Planck equations, perturbation theory, and data analysis for nonlinear stochastic systems with time delays. AB - We study nonlinear stochastic systems with time-delayed feedback using the concept of delay Fokker-Planck equations introduced by Guillouzic, L'Heureux, and Longtin. We derive an analytical expression for stationary distributions using first-order perturbation theory. We demonstrate how to determine drift functions and noise amplitudes of this kind of systems from experimental data. In addition, we show that the Fokker-Planck perspective for stochastic systems with time delays is consistent with the so-called extended phase-space approach to time delayed systems. PMID- 15903406 TI - Depletion kinetics in the photobleaching trapping reaction inside a flat microchannel. AB - The diffusion-limited kinetics of the growth of a depletion zone around a static point trap in a thin, long channel geometry was studied using a laser photobleaching experiment of fluorescein dye inside a flat rectangular capillary. The dynamics of the depletion zone was monitored by the theta distance, defined as the distance from the trap to the point where the reactant concentration has been locally depleted to the specified survival fraction (theta) of its initial bulk value. A dimensional crossover from two dimensions to one dimension, due to the finite width of the reaction zone, was observed. We define a "parallel" and a "perpendicular" theta distance, along the slab long and short dimensions, respectively, and study their time development as a means to study the asymmetrical nature of the slab geometry. For all theta values, the crossover occurs concurrently for both theta distances when the depletion zone touches the boundary for the first time. We derive theoretical expressions for this geometry and compare them with the experimental data. We also obtain important insight from the ratio of the reactant concentration profiles in the parallel and perpendicular directions. Exact enumeration and Monte Carlo simulations support the anomalous depletion scaling results. Nevertheless, the crossover time (tau(c)) is still found to scale with the width (W) of the rectangular reaction zone as tau(c) approximately W2 , as expected from the basic Einstein diffusion law. PMID- 15903407 TI - Numerical study of a binary Yukawa model in regimes characteristic of globular proteins in solutions. AB - The main goal of this paper is to assess the limits of validity, in the regime of low concentration and strong Coulomb coupling (high molecular charges), of a simple perturbative approximation to the radial distribution functions (RDF's), based upon a low-density expansion of the potential of mean force and proposed to describe protein-protein interactions in a recent small-angle-scattering (SAS) experimental study. A highly simplified Yukawa (screened Coulomb) model of monomers and dimers of a charged globular protein (beta-lactoglobulin) in solution is considered. We test the accuracy of the RDF approximation, as a necessary complementary part of the previous experimental investigation, by comparison with the fluid structure predicted by approximate integral equations and exact Monte Carlo (MC) simulations. In the MC calculations, an Ewald construction for Yukawa potentials has been used to take into account the long range part of the interactions in the weakly screened cases. Our results confirm that the perturbative first-order approximation is valid for this system even at strong Coulomb coupling, provided that the screening is not too weak (i.e., for Debye length smaller than monomer radius). A comparison of the MC results with integral equation calculations shows that both the hypernetted-chain (HNC) and Percus-Yevick closures have a satisfactory behavior under these regimes, with the HNC being superior throughout. The relevance of our findings for interpreting SAS results is also discussed. PMID- 15903408 TI - Heat capacity in a model polydisperse ferrofluid with narrow particle size distribution. AB - The influence of polydispersity on the phase equilibrium properties and the heat capacity of a dipolar system with additional short-range (repulsive + attractive) interactions (modeled by a shifted Lennard-Jones pair potential) is studied by means of a Monte Carlo scheme. The particle distribution of the investigated system is realized in the semigrand ensemble by tuning appropriately the underlying particle distribution density. The phase coexistence and heat capacity data are calculated with and without an applied magnetic field, and the obtained results are compared with the data determined in a monodisperse equivalent of the system. PMID- 15903409 TI - Stochastic resonance of electrochemical aperiodic spike trains. AB - Aperiodic stochastic resonance in an electrochemical system with excitable dynamics is characterized in experiments and simulations. Two different spike trains, one with stochastic and the other with chaotic interspike intervals, are imposed on the system as subthreshold aperiodic signals. Information transmission is quantified by the cross correlation between the subthreshold input signal and the noise induced system response. A maximum is exhibited in the input-output correlation as a function of the noise amplitude. Numerical simulations with an electrochemical model are in excellent agreement with the experimental observations. PMID- 15903410 TI - Two-dimensional two-state lattice-gas model. AB - We propose a two-dimensional lattice-gas (2D LG) model where atoms may be in two different states: the immobile state, in which they jump as usual in the LG model, and the running state, in which the atoms always jump in the driving direction. The model demonstrates a typical behavior of "traffic-jam" models: the system splits into domains of immobile atoms (jams) and running atoms. We considered four variants of the 2D LG model, namely the multilane and truly 2D models, each with "passive" and "active" atomic jumps. The model has the steady state with a power law distribution of jam sizes characterized by a universal exponent 3/2. The phase diagram of the model shows that the mobility of the 2D system is lower than the mobility of the 1D model due to the spreading of jams in the direction transverse to the driving direction. PMID- 15903411 TI - Phase velocity and phase diffusion in periodically driven discrete-state systems. AB - We develop a theory to calculate the effective phase diffusion coefficient and the mean phase velocity in periodically driven stochastic models with two discrete states. This theory is applied to a dichotomically driven Markovian two state system. Explicit expressions for the mean phase velocity, the effective phase diffusion coefficient, and the Pe clet number are analytically calculated. The latter indicates as a measure of phase-coherence forced synchronization of the stochastic system with respect to the periodic driving and exhibits a "bona fide" resonance. In a second step, the theory is applied to a non-Markovian two state system modeling excitable systems. The results prove again stochastic synchronization to the periodic driving and are in good agreement with simulations of a stochastic FitzHugh-Nagumo system. PMID- 15903412 TI - Brownian motion of finite-inertia particles in a simple shear flow. AB - Simultaneous diffusive and inertial motion of Brownian particles in laminar Couette flow is investigated via Lagrangian and Eulerian descriptions to determine the effect of particle inertia on diffusive transport in the long-time limit. The classical fluctuation dissipation theorem is used to calculate the amplitude of random-force correlations, thereby neglecting corrections of the order of the molecular relaxation time to the inverse shear rate. In the diffusive limit (time much greater than the particle relaxation time) the fluctuating particle-velocity autocorrelations functions are found to be stationary in time, the correlation in the streamwise direction being an exponential multiplied by an algebraic function and the cross correlation nonsymmetric in the time difference. The analytic, nonperturbative, evaluation of the particle-phase total pressure, which is calculated to be second order in the Stokes number (a dimensionless measure of particle inertia), shows that the particle phase behaves as a non-Newtonian fluid. The generalized Smoluchowski convective-diffusion equation, determined analytically from a combination of the particle-phase pressure tensor and the inertial acceleration term, contains a shear-dependent cross derivative term and an additional term along the streamwise direction, quadratic in the particle Stokes number. The long-time diffusion coefficients associated with the particle flux relative to the carrier flow are found to depend on particle inertia such that the streamwise diffusion coefficient becomes negative with increasing Stokes number, whereas one of the cross coefficients is always negative. The total diffusion coefficients measuring the rate of change of particle mean-square displacement are always positive as expected from general stability arguments. PMID- 15903413 TI - Satiated relative permeability of variable-aperture fractures. AB - Experimental studies of capillary-dominated displacements in variable-aperture fractures have demonstrated the occurrence of a satiated state at the end of invasion, where significant entrapment of the displaced phase occurs. The structure of this entrapped phase controls the behavior of flow and transport processes in the flowing phase. Recent studies have shown that the areal saturation of the flowing phase at satiation (S(f) ) is largely controlled by a single parameter C/delta , where C , the curvature number, weighs the mean in plane interfacial curvature relative to the mean out-of-plane interfacial curvature, and delta , the coefficient of variation of the aperture field, represents the strength of interface roughening induced by aperture variations. Here we consider the satiated relative permeability (k(rs)) to the flowing phase, which is defined as the relative permeability when the defending phase is fully entrapped. The satiated relative permeability is shown to be a well-defined function of S(f) over a wide range of C/delta , ranging from capillary fingering with significant entrapment (C/delta-->0) to smooth invasion with very little entrapment (C/delta > 1) . We develop a relationship for k(rs) as a function of S(f), by combining theoretical results for the effective permeability in a spatially correlated random permeability field, with results from continuum percolation theory for quantifying the influence of the entrapped phase. The resulting model for k(rs) also involves a dependence on delta . The predicted relative permeability values are accurate across the entire range of phase structures representative of capillary-dominated displacements in variable aperture fractures. PMID- 15903414 TI - Electrical conductivity of aqueous solutions of aluminum salts. AB - We present experimental measurements of the specific electrical conductivity (sigma) in aqueous solutions of aluminum salts at different temperatures, covering all salt concentrations from saturation to infinite dilution. The salts employed were AlCl(3), AlBr(3), AlI(3), and Al(NO(3))(3), which present a 1:3 relationship between the electrical charges of anion and cation. In addition, we have measured the density in all ranges of concentrations of the four aqueous electrolyte solutions at 298.15 K. The measured densities show an almost linear behavior with concentration, and we have fitted it to a second order polynomial with very high degree of approximation. The measurement of the specific conductivity at constant temperature reveals the existence of maxima in the conductivity vs concentration curves at molar concentrations around 1.5M for the three halide solutions studied, and at approximately 2M for the nitrate. We present a theoretical foundation for the existence of these maxima, based on the classical Debye-Hu ckel-Onsager hydrodynamic mean-field framework for electrical transport and its high concentration extensions, and also a brief consideration of ionic frictional coefficients using mode-coupling theory. We also found that the calculated values of the equivalent conductance vary in an approximately linear way with the square root of the concentration at concentrations as high as those where the maximum of sigma appears. Finally, and for completeness, we have measured the temperature dependence of the electrical conductivity at selected concentrations from 283 to 353 K, and performed a fit to an exponential equation of the Vogel-Fulcher-Tamman type. The values of the calculated temperatures of null mobility of the four salts are reported. PMID- 15903415 TI - Percolation of clusters with a residence time in the bond definition: Integral equation theory. AB - We consider the clustering and percolation of continuum systems whose particles interact via the Lennard-Jones pair potential. A cluster definition is used according to which two particles are considered directly connected (bonded) at time t if they remain within a distance d, the connectivity distance, during at least a time of duration tau, the residence time. An integral equation for the corresponding pair connectedness function, recently proposed by two of the authors [Phys. Rev. E 61, R6067 (2000)], is solved using the orthogonal polynomial approach developed by another of the authors [Phys. Rev. E 55, 426 (1997)]. We compare our results with those obtained by molecular dynamics simulations. PMID- 15903416 TI - Microscopic derivation of time-dependent density functional methods. AB - Time-dependent density functional methods (TDDFM) are studied from the microscopic viewpoint using projection operator methods in classical liquids. A density field is defined without averaging, so that a time evolution equation of the density field is derived with a random force. The derived equation includes a free energy functional, which is different from that defined in the TDDFM. The projection operator method provides the exact expression of the free energy functional. Another definition of the density field by an average leads to the equation of the TDDFM. In addition, an equation describing fluctuations is also derived. PMID- 15903417 TI - Inherent structures and Kauzmann temperature of confined liquids. AB - Calculations of the thermodynamical properties of a supercooled liquid confined in a matrix are performed with an inherent structure analysis. The liquid entropy is computed by means of a thermodynamical integration procedure. The contributions to the free energy of the liquid can be decoupled also in confinement in the configurational and the vibrational parts. We show that the vibrational entropy can be calculated in the harmonic approximation as in the bulk case. The Kauzmann temperature of the confined system is estimated from the behavior of the configurational entropy. PMID- 15903418 TI - Magnetoviscosity of semidilute ferrofluids and the role of dipolar interactions: comparison of molecular simulations and dynamical mean-field theory. AB - Extensive molecular simulations on a model ferrofluid are performed in order to study magnetoviscous and viscoelastic phenomena in semidilute ferrofluids. Simulation results of the nonequilibrium magnetization, shear viscosity, and normal stress differences are presented. Rotational and configurational contributions to the shear viscosity are analyzed and their influence on the magnetoviscous effect is discussed. The simplified model of noninteracting magnetic dipoles describes the nonequilibrium magnetization and the rotational viscosity, but does not account for configurational viscosity contributions and normal stress differences. Improved mean-field models that overcome these limitations show good agreement with the simulation results for weak dipolar interactions where the models should apply. Comparisons to simulation results for various interaction strengths allow us to determine the range of validity of the mean-field models. PMID- 15903419 TI - Short-ranged attractions in jammed liquids: how cooling can melt a glass. AB - We demonstrate that an extended picture of kinetic constraints in glass-forming liquids is sufficient to explain dynamic anomalies observed in dense suspensions of strongly attracting colloidal particles. We augment a simple model of heterogeneous relaxation with static attractions between facilitating excitations, in a way that mimics the structural effect of short-ranged interparticle attractions. The resulting spatial correlations among facilitated and unfacilitated regions give rise to relaxation mechanisms that account for nonmonotonic dependence of relaxation times on attraction strength as well as logarithmic decay of density correlations in time. These unusual features are a simple consequence of spatial segregation of kinetic constraints, suggesting an alternative physical perspective on attractive colloids than that suggested by mode-coupling theory. Based on the behavior of our model, we predict a crossover from super-Arrhenius to Arrhenius temperature dependence as attractions become dominant at fixed packing fraction. PMID- 15903420 TI - Behavior of granular materials under cyclic shear. AB - The design and development of a parallel plate shear cell for the study of large scale shear flows in granular materials is presented. The parallel plate geometry allows for shear studies without the effects of curvature found in the more common Couette experiments. A system of independently movable slats creates a well with side walls that deform in response to the motions of grains within the pack. This allows for true parallel plate shear with minimal interference from the containing geometry. The motions of the side walls also allow for a direct measurement of the velocity profile across the granular pack. Results are presented for applying this system to the study of transients in granular shear and for shear-induced crystallization. Initial shear profiles are found to vary from packing to packing, ranging from a linear profile across the entire system to an exponential decay with a width of approximately six bead diameters. As the system is sheared, the velocity profile becomes much sharper, resembling an exponential decay with a width of roughly three bead diameters. Further shearing produces velocity profiles which can no longer be fit to an exponential decay, but are better represented as a Gaussian decay or error function profile. Cyclic shear is found to produce large-scale ordering of the granular pack, which has a profound impact on the shear profile. There exist periods of time in which there is slipping between layers as well as periods of time in which the layered particles lock together resulting in very little relative motion. PMID- 15903421 TI - Simulations of vibrated granular medium with impact-velocity-dependent restitution coefficient. AB - We report numerical simulations of strongly vibrated granular materials designed to mimic recent experiments performed in both the presence and the absence of gravity. The coefficient of restitution used here depends on the impact velocity by taking into account both the viscoelastic and plastic deformations of particles, occurring at low and high velocities, respectively. We show that this model with impact-velocity-dependent restitution coefficient reproduces results that agree with experiments. We measure the scaling exponents of the granular temperature, collision frequency, impulse, and pressure with the vibrating piston velocity as the particle number increases. As the system changes from a homogeneous gas state at low density to a clustered state at high density, these exponents are all found to decrease continuously with increasing particle number. All these results differ significantly from classical inelastic hard sphere kinetic theory and previous simulations, both based on a constant restitution coefficient. PMID- 15903422 TI - Traveling waves in a water-immersed binary granular system vibrated within an annular cell. AB - It has been known since the time of Faraday that vertically vibrated fine grains may spontaneously form piles through their interaction with a fluid. More recently, it has been observed that a fine binary mixture may separate under vertical vibration through the differential influence of the fluid on the two granular components. Here, we report a detailed study of a system of water immersed bronze and glass grains held between two coaxial cylinders. Under vertical vibration, the bronze separates to form a layer above the glass, which itself breaks symmetry to form a pile. Symmetry is broken a second time by the bronze forming layers of different thicknesses upon the two slopes of the glass pile. The pile then travels as a wave with the thicker bronze layer upon its leading surface. We examine the conditions for these traveling waves and determine how their speed varies with particle size, frequency, and amplitude of vibration. A model is developed which provides a semiquantitative account of the wave motion. PMID- 15903423 TI - Creeping motion in granular flow. AB - The core of a quasi-two-dimensional rotating cylinder filled more than half full with glass beads rotates slightly faster than the cylinder itself and decreases in radius over time. Core precession depends linearly on the number of tumbler revolutions while core erosion varies logarithmically. Both processes serve to quantify the slow granular motion in the "fixed" bed and depend on the filling fraction and the tumbler rotation rate. A simple model, based on experimental observations of an exponential decrease in velocity parallel to the free surface, captures the primary features of the core dynamics. PMID- 15903424 TI - Dynamics of granular avalanches caused by local perturbations. AB - Surface flow of granular material is investigated within a continuum approach in two dimensions. The dynamics is described by the nonlinear coupling between a mobile layer and an erodible bed of static grains. Following previous studies, we use mass and momentum conservation to derive St. Venant-like equations for the evolution of the thickness R of the mobile layer and the profile Z of the bed. This approach allows the rheology in the flowing layer to be specified independently, and we consider in detail the two following models: a constant plug flow and a linear velocity profile. We study and compare these models for nonstationary avalanches triggered by a localized amount of mobile grains on a bed of initially constant slope. We solve analytically the nonlinear dynamical equations by the method of characteristics. This enables us to investigate the temporal evolution of the avalanche size, amplitude, and shape as a function of model parameters and initial conditions. In particular, we can compute their large time behavior as well as the condition for the formation of shocks. PMID- 15903425 TI - Concentration and velocity patterns in a horizontal rotating suspension of non Brownian settling particles. AB - We report a systematic experimental study of concentration and velocity patterns formed in a horizontal rotating cylinder filled completely with a monodisperse suspension of non-Brownian settling particles. The system shows a series of concentration and velocity patterns, or phases, with varying rotation rate and solvent viscosity. Individual phases are studied using both side and cross sectional imaging to examine the detailed flow structures. The overall phase diagram of the system is mapped out as a function of the rotation rate and solvent viscosity. Attempts are made to analyze the functional form of the phase boundaries in order to understand the transition mechanism between different phases. PMID- 15903426 TI - Rigidity percolation in dispersions with a structured viscoelastic matrix. AB - This paper deals with rigidity percolation in composite materials consisting of a dispersion of mineral particles in a microstructured viscoelastic matrix. The viscoelastic matrix in this specific case is a hydrocarbon refinery residue. In a set of model random composites the mean interparticle surface-to-surface distance was controlled, changing particle volume fraction phi and particle number density independently. This was achieved by mixing two sets of monodisperse particles with widely differing radii (0.35 microm and 17.5 microm) with the matrix. A scaling exponent of 3.9 +/- 0.6 for the storage modulus G' vs phi- phi(c) was observed above a threshold phi(c) , in good agreement with theoretical values for rigidity percolation. It is found that at the rigidity-percolation threshold the pore structure, as characterized by the mean surface-to-surface distance for the filler, rather than the filler volume fraction, is similar for different types of composites. This behavior is explained from the internal structure of the viscoelastic matrix, which consists of fractal solid aggregates dissolved in a viscous medium; the effective radius of these aggregates and the mean surface-to surface distance together determine whether or not the aggregates are capable of providing rigidity to the composite. The explanation is further supported by a qualitative comparison with effective-medium calculations. These indicate that the observed breakdown of time-temperature superposition near phi(c) is due to the appearance of a time scale characteristic for the mechanical interplay between the viscous binder phase and the purely elastic solid particles. PMID- 15903427 TI - Two-dimensional flow of foam around an obstacle: force measurements. AB - A Stokes experiment for foams is proposed. It consists of a two-dimensional flow of a foam, confined between a water subphase and a top plate, around a fixed circular obstacle. We present systematic measurements of the drag exerted by the flowing foam on the obstacle versus various separately controlled parameters: flow rate, bubble volume, bulk viscosity, obstacle size, shape, and boundary conditions. We separate the drag into two contributions: an elastic one (yield drag) at vanishing flow rate and a fluid one (viscous coefficient) increasing with flow rate. We quantify the influence of each control parameter on the drag. The results exhibit in particular a power-law dependence of the drag as a function of the bulk viscosity and the flow rate with two different exponents. Moreover, we show that the drag decreases with bubble size and increases proportionally to the obstacle size. We quantify the effect of shape through a dimensional drag coefficient, and we show that the effect of boundary conditions is small. PMID- 15903428 TI - Anisotropic mean-square displacements in two-dimensional colloidal crystals of tilted dipoles. AB - Superparamagnetic colloidal particles confined to a flat horizontal air-water interface in an external magnetic field, which is tilted relative to the interface, form anisotropic two-dimensional crystals resulting from their mutual dipole-dipole interactions. Using real-space experiments and harmonic lattice theory we explore the mean-square displacements of the particles in the directions parallel and perpendicular to the in-plane component of the external magnetic field as a function of the tilt angle. We find that the anisotropy of the mean-square displacement behaves nonmonotonically as a function of the tilt angle and does not correlate with the structural anisotropy of the crystal. PMID- 15903429 TI - Tilt grain boundary instabilities in three-dimensional lamellar patterns. AB - We identify a finite wave-number instability of a 90 degrees tilt grain boundary in three-dimensional lamellar phases which is absent in two-dimensional configurations. Both a stability analysis of the slowly varying amplitude or envelope equation for the boundary, and a direct numerical solution of an order parameter model equation are presented. The instability mode involves two dimensional perturbations of the planar base boundary, and is suppressed for purely one-dimensional perturbations. We find that both the most unstable wave numbers and their growth rate increase with epsilon , the dimensionless distance away from threshold of the lamellar phase. PMID- 15903430 TI - Equilibrium polymerization in the Stockmayer fluid as a model of supermolecular self-organization. AB - A diverse range of molecular self-organization processes arises from a competition between directional and isotropic van der Waals intermolecular interactions. We conduct Monte Carlo simulations of the Stockmayer fluid (SF) with a large dipolar interaction as a minimal self-organization model and focus on basic thermodynamic properties that are needed to characterize the polymerization transition that occurs in this fluid. In particular, we determine the polymerization transition lines from the maximum in the specific heat, C(v), and the inflection point in the extent of polymerization, Phi. We also characterize the geometry (radius of gyration R(g), chain length L, chain topology) of the clusters that form in this associating fluid as a function of temperature, T, and concentration, rho . The pressure, P, and the second virial coefficient, B2, were determined, since these properties contain essential information about the strength of the isotropic (van der Waals) interactions. Our simulations indicate that the locations of the polymerization lines are quantitatively consistent with a model of equilibrium polymerization with the enthalpy of polymerization ("sticking energy") fixed by the minimum in the intermolecular potential. The polymerization transition in the SF is accompanied by a topological transition from predominantly linear to ring polymers upon cooling that is driven by the minimization of the dipolar energy of the clusters. We also find that the basic interaction parameters describing polymerization and phase separation in the SF can be estimated based on the existing theory of equilibrium polymerization, but the theory must be refined to account for ring formation in order to accurately describe the configurational properties of this model self-organizing fluid. PMID- 15903431 TI - Method to calculate electrical forces acting on a sphere in an electrorheological fluid. AB - We describe a method to calculate the electrical force acting on a sphere in a suspension of dielectric spheres in a host with a different dielectric constant, under the assumption that a spatially uniform electric field is applied. The method uses a spectral representation for the total electrostatic energy of the composite. The force is expressed as a certain gradient of this energy, which can be expressed in a closed analytic form rather than evaluated as a numerical derivative. The method is applicable even when both the spheres and the host have frequency-dependent dielectric functions and nonzero conductivities, provided the system is in the quasistatic regime. In principle, it includes all multipolar contributions to the force, and it can be used to calculate multibody as well as pairwise forces. We also present several numerical examples, including host fluids with finite conductivities. The force between spheres approaches the dipole-dipole limit, as expected, at large separations, but departs drastically from that limit when the spheres are nearly in contact. The force may also change sign as a function of frequency when the host is a slightly conducting fluid. PMID- 15903432 TI - Liquid polymorphism and density anomaly in a lattice gas model. AB - We present a simple model for an associating liquid in which polymorphism and density anomaly are connected. Our model combines a two dimensional lattice gas with particles interacting through a soft core potential and orientational degrees of freedom represented through thermal "ice variables." The competition between the directional attractive forces and the soft core potential leads to a phase diagram in which two liquid phases and a density anomaly are present. The coexistence line between the low density liquid and the high density liquid has a positive slope contradicting the surmise that the presence of a density anomaly implies that the high density liquid is more entropic than the low density liquid. PMID- 15903433 TI - Heterogeneous dynamics on the microsecond scale in simulated Ni0.5 Zr0.5 metallic melts far below the glass temperature. AB - From molecular dynamics simulations for up to 2.5 micros, results are reported on structural dynamics in glass-forming metallic Ni(0.5) Zr(0.5) melts at 700 and 785 K. After elimination of high-frequency fluctuations, cascades of atoms and avalanches of atom chains, local in space and time, are identified as complex processes that govern the heterogeneous low-frequency dynamics in sufficiently aged structures near the glass temperature. PMID- 15903434 TI - Glassy behavior of a percolative water-protein system. AB - We show that is possible to look at the glass transition as a percolation transition in phase space. This study has been carried out on a hydrated globular enzyme for which the thermodynamic transition and the percolative transition could have a functional significance. The approach adopted is based on the identity of roles played respectively by the glass transition temperature T(o) and the critical hydration threshold h(c) for the percolation of protons on the surface and through the protein, given that dynamical arrest is observed at temperatures and hydration below T(o) and h(c). Theoretical predictions for temperature dependence of the nonexponentiality parameter, beta(KWW) , appearing in the KWW relaxation function, indicate that at high temperatures, beta(KWW) remains insensitive to temperature changes, whereas in the vicinity of the glass transition, beta(KWW) is linearly increasing with temperature. The low temperature limit of beta(KWW) is about 1/3 and its temperature-independent behavior starts at 1.23 T(g): both predictions have been verified in the present study. PMID- 15903435 TI - Dynamics in a two-dimensional core-softened fluid. AB - The dynamical properties of a model one-component core-softened fluid with purely repulsive interactions are found to be very complex. At low temperature the fluid structure exhibits cluster motifs including dimers, stripes, and polygons, depending on density. Single-particle diffusion and the velocity, shear-stress, and wave-vector-dependent current correlation functions have all been calculated using molecular dynamics simulations. The results highlight the presence of well resolved single-particle and collective motions, which is remarkable for what is essentially a "simple" one-component fluid. PMID- 15903436 TI - Free-volume dynamics in glasses and supercooled liquids. AB - A free-volume theory is developed based on the defect diffusion model (DDM). In addition, positronium annihilation lifetime spectroscopy (PALS) ortho-positronium free-volume and intensity data are presented for poly(propylene glycol) with a molecular weight of 4000 (PPG 4000) in both the glassy and liquid states and dielectric relaxation and electrical conductivity data are reported for PPG 4000 in the liquid state. The DDM is used to interpret all of the data for PPG 4000 and previously reported PALS and dielectric relaxation data for glycerol. It is shown that while the PPG 4000 data exhibit a preference for the three-halves power law, the data for glycerol favor the first power (standard Vogel-Fulcher Tammann) law. Good agreement between the DDM and the experimental results is found for all of the electrical data and the PALS free-volume data. While reasonable agreement is also found for the PALS intensity data for PPG 4000, a discrepancy exists between the experimental PALS intensity data and theory for glycerol. For the electrical conductivity for PPG 4000, a transition is observed at the same temperature (about 1.4 T(g) where T(g) is the glass transition temperature) where the PALS free volume changes from steeply rising with temperature to approximately independent of temperature. The same behavior is observed at about 1.5 T(g) for previously reported dielectric relaxation and PALS data for glycerol. Model parameters are presented that show the dominance of mobile single defects above (1.4-1.5) T(g) and the dominance of immobile clustered single defects below T(g) . Finally, a coherent picture of glasses and glass-forming liquids is presented based on the theory and results of the experiments. PMID- 15903437 TI - Nucleation times in the two-dimensional Ising model. AB - We consider the distribution of nucleation times in systems with Brownian type dynamics, as described by classical nucleation theory. This is studied for a prototype system: the two-dimensional Ising model with spin-flip dynamics in an external magnetic field. Direct simulation results for the nucleation times, spanning more than four orders of magnitude, are compared with theoretical predictions. In contrast to usual treatments we determine size-dependent droplet free energies and effective transition rates for growth and shrinkage directly from our simulations. The free energies so determined are well described by the classical Becker-Do ring expression, provided one uses an effective surface tension that exceeds the macroscopic surface tension by up to 20%. Within this framework there is good agreement between simulation results and theoretical predictions for the mean nucleation time. In addition we consider the short-time behavior of the nucleation probability after an initial quench into the metastable state. We present theoretical estimates and show that these too agree well with simulation results. PMID- 15903438 TI - Side-branch growth in two-dimensional dendrites. I. Experiments. AB - The dynamics of growth of dendrites' side branches is investigated experimentally during the crystallization of solutions of ammonium bromide in a quasi-two dimensional cell. Two regimes are observed. At small values of the Peclet number a self-affine fractal forms. In this regime it is known that the mean lateral front grows as t(0.5). Here the length of each individual branch is shown to grow (before being screened off) with a power-law behavior t (alpha(n)). The value of the exponent alpha(n) (0.5< or = alpha(n) < or =1) is determined from the start by the strength of the initial disturbance. Coarsening then takes place, when the branches of small alpha(n) are screened off by their neighbors. The corresponding decay of the growth of a weak branch is exponential and defined by its geometrical position relative to its dominant neighbors. These results show that the branch structure results from a deterministic growth of initially random disturbances. At large values of the Peclet number, the faster of the side branches escape and become independent dendrites. The global structure then covers a finite fraction of the two-dimensional space. The crossover between the two regimes and the spacing of these independent branches are characterized. PMID- 15903439 TI - Decay of metastable phases in a model for the catalytic oxidation of CO. AB - We study by kinetic Monte Carlo simulations the dynamic behavior of a Ziff-Gulari Barshad model with CO desorption for the reaction CO + O-->CO(2) on a catalytic surface. Finite-size scaling analysis of the fluctuations and the fourth-order order-parameter cumulant show that below a critical CO desorption rate, the model exhibits a nonequilibrium first-order phase transition between low and high CO coverage phases. We calculate several points on the coexistence curve. We also measure the metastable lifetimes associated with the transition from the low CO coverage phase to the high CO coverage phase, and vice versa. Our results indicate that the transition process follows a mechanism very similar to the decay of metastable phases associated with equilibrium first-order phase transitions and can be described by the classic Kolmogorov-Johnson-Mehl-Avrami theory of phase transformation by nucleation and growth. In the present case, the desorption parameter plays the role of temperature, and the distance to the coexistence curve plays the role of an external field or supersaturation. We identify two distinct regimes, depending on whether the system is far from or close to the coexistence curve, in which the statistical properties and the system-size dependence of the lifetimes are different, corresponding to multidroplet or single-droplet decay, respectively. The crossover between the two regimes approaches the coexistence curve logarithmically with system size, analogous to the behavior of the crossover between multidroplet and single droplet metastable decay near an equilibrium first-order phase transition. PMID- 15903440 TI - Exact solution of a one-dimensional model of strained epitaxy on a periodically modulated substrate. AB - We consider a one-dimensional lattice gas model of strained epitaxy with the elastic strain accounted for through a finite number of cluster interactions comprising contiguous atomic chains. Interactions of this type arise in the models of strained epitaxy based on the Frenkel-Kontorova model. Furthermore, the deposited atoms interact with the substrate via an arbitrary periodic potential of period L. This model is solved exactly with the use of an appropriately adopted technique developed recently in the theory of protein folding. The advantage of the proposed approach over the standard transfer-matrix method is that it reduces the problem to finding the largest eigenvalue of a matrix of size L instead of 2(L-1), which is vital in the case of nanostructures where L may measure in hundreds of interatomic distances. Our major conclusion is that the substrate modulation always facilitates the size calibration of self-assembled nanoparticles in one- and two-dimensional systems. PMID- 15903441 TI - Phase equilibria in solutions of platelike particles: systems with steric and dispersive interactions between the platelets. AB - Our statistical thermodynamics model of solution of stiff, platelike, biaxial particles interacting solely via repulsion on contact (athermal limit) [Phys. Rev. E 62, 5011 (2000)] is extended to incorporate dispersion interactions between the particles. Dispersion forces between anisotropic particles are accounted for using the Imura-Okano approach. Numerical calculations specialized to solutions of either rods or disks show that besides the isotropic-nematic biphasic coexistence range, inclusion of attractive forces resulted in the appearance of nematic-nematic coexistence in both, disks and rods, solutions. The critical divergence of the difference between the order parameters and concentrations of the two nematics is observed while approaching the critical temperature. The minimum aspect ratio of rods or disks for the formation of the nematic phase is also discussed. PMID- 15903442 TI - Surface-induced orientational order in stretched nanoscale-sized polymer dispersed liquid-crystal droplets. AB - We investigate orientational ordering in stretched polymer-dispersed liquid crystal (PDLC) droplets using deuterium nuclear magnetic resonance, in the nematic and isotropic phases. In the latter case, we estimate the surface order parameter S(0) and the thickness of the interfacial layer from the temperature independent surface ordering model for an elliptical cavity with a varying aspect ratio. A simple phenomenological model well describes the quadrupole splitting frequency of NMR spectra in the isotropic phase. The strain dependence of S(0) suggests that stretching-induced changes in the orientation of polymer chains in the PDLC matrix noticeably affect liquid-crystal surface anchoring. Experimental results are supported by simulated NMR spectra obtained as output from Monte Carlo simulations of paranematic ordering in ellipsoidal droplets based on the Lebwohl-Lasher lattice model. PMID- 15903443 TI - Numerical study of a calamitic liquid-crystal model: phase behavior and structure. AB - We have studied an idealized calamitic liquid-crystal model, consisting of a linear rigid array of nine soft repulsive spheres, employing both theory and molecular dynamics simulation. The phase behavior (which includes crystalline, smectic, nematic, and isotropic phases) and structure of a collection of these rodlike particles have been determined by molecular dynamics simulation in an isothermal-isobaric ensemble. The liquid crystalline part of the phase diagram has been compared to that emerging from an Onsager-type density-functional theory. We have found a fair agreement between theory and computer simulation results, with a similar accuracy for the smectic to nematic and nematic to isotropic phase transitions. PMID- 15903444 TI - Twist of cholesteric liquid crystal cells: stability of helical structures and anchoring energy effects. AB - We consider helical configurations of a cholesteric liquid crystal (CLC) sandwiched between two substrates with homogeneous director orientation favored at both confining plates. We study the CLC twist wave number q characterizing the helical structures in relation to the free twisting number q(0) which determines the equilibrium value of CLC pitch P(0) = 2 pi/ q(0) . We investigate the instability mechanism underlying transitions between helical structures with different spiral half-turn numbers. Stability analysis shows that for equal finite anchoring strengths this mechanism can be dominated by in-plane director fluctuations. In this case the metastable helical configurations are separated by the energy barriers and the transitions can be described as the director slippage through these barriers. We extend our analysis to the case of an asymmetric CLC cell in which the anchoring strengths at the two substrates are different. The asymmetry introduces two qualitatively different effects: (a) the intervals of twist wave numbers representing locally stable configurations with adjacent helix half-turn numbers are now separated by instability gaps; and (b) sufficiently large asymmetry, when the difference between azimuthal anchoring extrapolation lengths exceeds the thickness of the cell, will suppress the jumplike behavior of the twist wave number. PMID- 15903445 TI - Structure and mechanical properties of liquid crystalline filaments. AB - The formation of stable freely suspended filaments is an interesting peculiarity of some liquid crystal phases. So far, little is known about their structure and stability. Similarly to free-standing smectic films, an internal molecular structure of the mesophase stabilizes these macroscopically well-ordered objects with length to diameter ratios of 10(3) and above. In this paper, we report observations of smectic liquid crystal fibers formed by bent-shaped molecules in different mesophases. Our study, employing several experimental techniques, focuses on mechanical and structural aspects of fiber formation such as internal structure, stability, and mechanical and optical properties. PMID- 15903446 TI - Calorimetric investigations of liquid-crystal compounds exhibiting almost no layer-shrinkage behavior through the smectic-A-smectic- C* transition. AB - Heat-capacity measurements have been made on liquid-crystal compounds exhibiting almost no layer-shrinkage (NLS) behavior through the Sm-A-Sm- C(*) phase transition. The transition was found to be second order for two of the substances studied. It was found that the heat-capacity anomaly accompanying a second-order Sm-A-Sm- C(*) transition with NLS behavior is quite similar to that observed for typical antiferroelectric liquid crystals of the 4-(1 methylheptyloxycarbonyl)phenyl 4'-octyloxybiphenyl-4-carboxylate (MHPOBC) group, showing three-dimensional (3D) XY behavior in the vicinity of the transition. On the other hand, for one compound which shows a weakly first-order transition, the anomaly is almost symmetric above and below T(c) , with a significant fluctuation effect in the Sm-A phase. For this compound, the critical behavior of the heat capacity anomaly is almost tricritical in the immediate vicinity of T(c) , while away from T(c) the behavior can be explained with the 3D XY model. This suggests that the underlying transition with the 3D XY critical behavior is driven to almost being tricritical but remaining weakly first order. No indication of low dimensional character in the critical behavior was found in both cases. PMID- 15903447 TI - Fluctuations of the tensor order-parameter modes in a cholesteric liquid crystal. AB - We use a Landau-de Gennes free energy to calculate the fluctuations of the five independent modes of the tensor order parameter for a cholesteric liquid crystal. Our results include, as a limiting case, the two classical director modes, known as the twist mode and the "umbrella" mode. We find, however, in contrast to the classical director model, that there can be substantial temperature dependence to the umbrella mode, as well as three additional modes near the transition to the isotropic phase. We comment on a recent experiment that suggests that two of these additional modes may have already been detected. PMID- 15903448 TI - Branched polymers on the Given-Mandelbrot family of fractals. AB - We study the average number A (n) per site of the number of different configurations of a branched polymer of n bonds on the Given-Mandelbrot family of fractals using exact real-space renormalization. Different members of the family are characterized by an integer parameter b , 2 < or = b < or = infinity . The fractal dimension varies from log(2) 3 to 2 as b is varied from 2 to infinity. We find that for all b > or = 3 , A (n) varies as lambda(n) exp (b n(psi)) where lambda and b are some constants, and 0 < psi < 1 . We determine the exponent psi, and the size exponent nu (average diameter of polymer varies as n(nu) ), exactly for all b , 3 < or = b < o r= infinity . This generalizes the earlier results of Knezevic and Vannimenus for b = 3 [Phys. Rev B 35, 4988 (1987)]. PMID- 15903449 TI - Semiflexible polymer condensates in poor solvents: toroid versus spherical geometries. AB - Semiflexible polymers, such as DNA, in the presence of a condensing agent often form toroids. This is due to a balance between bending and surface area free energy penalties. Here we show why in experiments all the toroids have been found to have similar physical size. We also introduce a novel morphology, that of the hollow sphere, which is favored for long polymer chains. This offers the possibility of encapsulating material inside a "vesicle" made of semiflexible polymers. We also consider the case of many such polymer chains placed in a poor solvent. We show a transition between two morphologies occur on increasing concentration of polymer chains, from a thickened toroid to a spherical globule. PMID- 15903450 TI - End-to-end distribution function of stiff polymers for all persistence lengths. AB - We set up recursion relations for calculating all even moments of the end-to-end distance of Porod-Kratky wormlike chains in D dimensions. From these moments we derive a simple analytic expression for the end-to-end distribution in three dimensions valid for all peristence lengths. It is in excellent agreement with Monte Carlo data for stiff chains and approaches the Gaussian random-walk distributions for low stiffness. PMID- 15903451 TI - Anisotropic suppression of phase separation in polymer solutions by oscillatory shear. AB - We consider the effects of subjecting a polymer solution to a simultaneous oscillatory shear flow and temperature jump into the two-phase region of the phase diagram. We predict that if the oscillatory shear stresses are significant enough then the flow suppresses phase separation in the flow direction, leading to the possibility of creating strongly aligned structures. We construct a quantitative dynamic phase diagram in the amplitude-frequency plane showing the conditions for the growth or decay of concentration fluctuations. Further, we discuss the time dependence of periodic structure factors in the flow-vorticity and flow-gradient planes. It is also shown that significantly enhanced scattering occurs at large scattering vectors regardless of whether the unbounded growth of fluctuations at smaller scattering vectors is suppressed. PMID- 15903452 TI - Effect of surface undulation on polymer adsorption. AB - We study the adsorption of a long, flexible polymer (ideal or self-avoiding chain) interacting with a rough surface via a finite-range attraction. Within the Edwards equation approach, we develop a variational method to find the segmental distribution and the free energy of an adsorbed chain. As adsorption becomes strong, the segments tend to be localized within the valleys rather than above the hills of the undulating surface, resulting in a decrease of adsorption thickness. Consequently, the surface undulation enhances adsorption in the case of a strongly adsorbed chain whereas the undulation suppresses it for a weakly adsorbed one, since the enhanced entropic repulsion is dominant over the attraction from the surface. Considering the surface with undulation characterized by a Gaussian correlation as an example, we find an optimal correlation length at which the adsorption becomes the strongest and a critical correlation length below which desorption is induced. PMID- 15903453 TI - Characterization of degree frequency distribution in protein interaction networks. AB - In this work, we analyze the degree frequency distribution in the yeast protein interaction network by studying a previously proposed duplication network model. This model correctly predicts the observed degree distribution (a power law for large degree values and a departure from this behavior for small degree). We numerically and analytically characterize this distribution as a mixture of random and power-law behavior, and make a comparative study of the robustness of the network model against realistic perturbations. We conclude that the particular distribution observed in both the model and the experimental data has many advantages in terms of dynamical and topological robustness and could have emerged in the evolutionary history as a sort of compromise between purely deterministic and random underlying mechanisms of network growth. PMID- 15903454 TI - Understanding mechanochemical coupling in kinesins using first-passage-time processes. AB - Kinesins are processive motor proteins that move along microtubules in a stepwise manner, and their motion is powered by the hydrolysis of ATP. Recent experiments have investigated the coupling between the individual steps of single kinesin molecules and ATP hydrolysis, taking explicitly into account forward steps, backward steps, and detachments. A theoretical study of mechanochemical coupling in kinesins, which extends the approach used successfully to describe the dynamics of motor proteins, is presented. The possibility of irreversible detachments of kinesins from the microtubules is explicitly taken into account. Using the method of first-passage times, experimental data on the mechanochemical coupling in kinesins are fully described using the simplest two-state model. It is shown that the dwell times for the kinesin to move one step forward or backward, or to dissociate irreversibly, are the same, although the probabilities of these events are different. It is concluded that the current theoretical view that only the forward motion of the motor protein molecule is coupled to ATP hydrolysis--is consistent with all available experimental observations for kinesins. PMID- 15903455 TI - Selective advantage for conservative viruses. AB - In this article we study the full semiconservative treatment of a model for the coevolution of a virus and an adaptive immune system. Regions of viability are calculated for both conservatively and semiconservatively replicating viruses interacting with a realistic semiconservatively replicating immune system. The conservative virus is found to have a selective advantage in the form of an ability to survive in regions with a wider range of mutation rates than its semiconservative counterpart, as well as an increased replication rate where both species can survive. This may help explain the existence of a rich range of viruses with conservatively replicating genomes, a trait that is found nowhere else in nature. PMID- 15903456 TI - Discrete layers of interacting growing protein seeds: convective and morphological stages of evolution. AB - The growth of several macromolecular seeds uniformly distributed on the bottom of a protein reactor (i.e., a discrete layer of N crystals embedded within a horizontal layer of liquid with no-slip boundaries) under microgravity conditions is investigated for different values of N and for two values of the geometrical aspect ratio of the container. The fluid dynamics of the growth reactor and the morphological (shape-change) evolution of the crystals are analyzed by means of a recently developed moving boundary method based on differential equations coming from the protein "surface incorporation kinetics." The face growth rates are found to depend on the complex multicellular structure of the convective field and on associated "pluming phenomena." This correspondence is indirect evidence of the fact that mass transport in the bulk and surface attachment kinetics are competitive as rate-limiting steps for growth. Significant adjustments in the roll pattern take place as time passes. The convective field undergoes an interesting sequence of transitions to different values of the mode and to different numbers of rising solutal jets. The structure of the velocity field and the solutal effects, in turn, exhibit sensitivity to the number of interacting crystals if this number is small. In the opposite case, a certain degree of periodicity can be highlighted for a core zone not affected by edge effects. The results with no-slip lateral walls are compared with those for periodic boundary conditions to assess the role played by geometrical constraints in determining edge effects and the wavelength selection process. The numerical method provides "microscopic" and "morphological" details as well as general rules and trends about the macroscopic evolution (i.e., "ensemble behaviors") of the system. PMID- 15903457 TI - Nonlinear elasticity of an alpha -helical polypeptide. AB - We study a minimal extension of the wormlike chain model to describe polypeptides having alpha-helical secondary structure. In this model the presence or absence of secondary structure enters as a scalar variable that controls the local chain bending modulus. Using this model we compute the extensional compliance of an alpha-helix under tensile stress, the bending compliance of the molecule under externally imposed torques, and the nonlinear interaction of such torques and forces on the molecule. We find that, due to coupling of the "internal" secondary structure variables to the conformational degrees of freedom of the polymer, the molecule has a highly nonlinear response to applied stress and force couples. In particular we demonstrate a sharp lengthening transition under applied force and a buckling transition under applied torque. Finally, we speculate that the inherent bistability of the molecule may underlie protein conformational change in vivo. PMID- 15903458 TI - Multicanonical study of coarse-grained off-lattice models for folding heteropolymers. AB - We have performed multicanonical simulations of hydrophobic-hydrophilic heteropolymers with two simple effective, coarse-grained off-lattice models to study the influence of specific interactions in the models on conformational transitions of selected sequences with 20 monomers. Another aspect of the investigation was the comparison with the purely hydrophobic homopolymer and the study of general conformational properties induced by the "disorder" in the sequence of a heteropolymer. Furthermore, we applied an optimization algorithm to sequences with up to 55 monomers and compared the global-energy minimum found with lowest-energy states identified within the multicanonical simulation. This was used to find out how reliable the multicanonical method samples the free energy landscape, in particular for low temperatures. PMID- 15903459 TI - Nonequilibrium model for estimating parameters of deleterious mutations. AB - Deleterious mutations are of extreme evolutionary importance because, even though they are eliminated by natural selection, their continuous pressure creates a pool of variability in natural populations. They are of potential relevance for the existence of several features in evolution, such as sexual reproduction, and pose a risk to small asexual populations. Despite their extreme importance, the deleterious mutation rate and the effects of each mutation on fitness are poorly known quantities. Here we analyze a simple model that can be applied to simple experiments, in microorganisms, aiming at the quantification of these values. PMID- 15903460 TI - Reaction-diffusion model for pattern formation in E. coli swarming colonies with slime. AB - A new experimental colonial pattern and pattern transition observed in E. coli MG1655 swarming cells grown on semisolid agar are described. We present a reaction-diffusion model that, taking into account the slime generated by these cells and its influence on the bacterial differentiation and motion, reproduces the pattern and successfully predicts the observed changes when the colonial collective motility is limited. In spite of having small nonhyperflagellated swarming cells, under these experimental conditions E. coli MG1655 can very rapidly colonize a surface, with a low branching rate, thanks to a strong fluid production and a locally incremented density of motile, lubricating cells. PMID- 15903461 TI - Scaling of horizontal and vertical fixational eye movements. AB - Eye movements during fixation of a stationary target prevent the adaptation of the visual system to continuous illumination and inhibit fading of the image. These random, involuntary, small movements are restricted at long time scales so as to keep the target at the center of the field of view. Here we use detrended fluctuation analysis in order to study the properties of fixational eye movements at different time scales. Results show different scaling behavior between horizontal and vertical movements. When the small ballistic movements, i.e., microsaccades, are removed, the scaling exponents in both planes become similar. Our findings suggest that microsaccades enhance the persistence at short time scales mostly in the horizontal component and much less in the vertical component. This difference may be due to the need for continuously moving the eyes in the horizontal plane, in order to match the stereoscopic image for different viewing distances. PMID- 15903462 TI - How the chromatin fiber deals with topological constraints. AB - In the nuclei of eukaryotic cells, DNA is packaged through several levels of compaction in an orderly retrievable way that enables the correct regulation of gene expression. The functional dynamics of this assembly involves the unwinding of the so-called 30-nm chromatin fiber and accordingly imposes strong topological constraints. We present a general method for computing both the twist and the writhe of any winding pattern. An explicit derivation is implemented for the chromatin fiber which provides the linking number of DNA in eukaryotic chromosomes. We show that there exists one and only one unwinding path which satisfies both topological and mechanical constraints that DNA has to deal with during condensation/decondensation processes. PMID- 15903463 TI - Phonon spectra and thermodynamic properties of the infinite polyalanine alpha helix: a density-functional-theory-based harmonic vibrational analysis. AB - We have performed a density-functional theory harmonic vibrational analysis of the infinite polyalanine alpha helix. The calculated phonon dispersion spectrum shows excellent agreement to available experimental data, except for the high frequency hydrogen stretching modes which show characteristic shifts due to anharmonic effects. A major advantage compared to previously performed empirical force field studies is that long range effects such as electrostatic interaction and polarization are intrinsically taken into account for characterizing hydrogen bond formation in the helix. Our results indicate that these effects are crucial to accurately describe the low frequency acoustical branches and lead to a significantly better agreement with experiment for the specific heat in the low temperature range. PMID- 15903464 TI - Quantitative point source photoacoustic inversion formulas for scattering and absorbing media. AB - We present here an expression for the photoacoustic contribution of an optical point source in a diffusive and absorbing medium. By using this measurement as a reference, we present a direct inversion formula that recovers the absorption map quantitatively, at the same time accounting for instrumental factors such as the source strength, the shape of the optical pulse, and the impulse response and finite size of the transducers. We further validate this expression through accurate numerical simulations showing that the absorption map is recovered quantitatively in the presence of a rotating geometry. We finally discuss how the presented solutions for point sources within the photoacoustic problem enable the use of concurrent fluorescence and ultrasound measurements as appropriate for a hybrid tomographic system. The proposed system could retrieve absorption information using photoacoustic measurements, and use these data to more accurately describe the fluorescence problem and improve reconstruction fidelity. PMID- 15903465 TI - Evidence on DNA slippage step-length distribution. AB - A simple model based on a master equation is constructed in order to reveal the details of the mutational events modifying simple sequence repeats in the human genome, A database of simple repeats together with their flanking sequences comprising approximately 10(5) entries from all 24 human chromosomes was constructed. By aligning the pairs of fragments of sequences containing the repeat elements, the matrices that count the number of slippage events were evaluated. The counts were then used as a target to be reproduced by our theoretical model, in which the elongation and shortening of the repeats proceed through a mechanism in which the step lengths exhibit a decaying distribution in the form of an inverse power law rather than through one nucleotide extension or deletion, which was the most frequent supposition in previous studies. PMID- 15903466 TI - Simulations of transient membrane behavior in cells subjected to a high-intensity ultrashort electric pulse. AB - A molecular dynamics (MD) scheme is combined with a distributed circuit model for a self-consistent analysis of the transient membrane response for cells subjected to an ultrashort (nanosecond) high-intensity (approximately 0.01-V/nm spatially averaged field) voltage pulse. The dynamical, stochastic, many-body aspects are treated at the molecular level by resorting to a course-grained representation of the membrane lipid molecules. Coupling the Smoluchowski equation to the distributed electrical model for current flow provides the time-dependent transmembrane fields for the MD simulations. A good match between the simulation results and available experimental data is obtained. Predictions include pore formation times of about 5-6 ns. It is also shown that the pore formation process would tend to begin from the anodic side of an electrically stressed membrane. Furthermore, the present simulations demonstrate that ions could facilitate pore formation. This could be of practical importance and have direct relevance to the recent observations of calcium release from the endoplasmic reticulum in cells subjected to such ultrashort, high-intensity pulses. PMID- 15903467 TI - Fractal scaling of microbial colonies affects growth. AB - The growth dynamics of filamentary microbial colonies is investigated. Fractality of the fungal or actinomycetes colonies is shown both theoretically and in numerical experiments to play an important role. The growth observed in real colonies is described by the assumption of time-dependent fractality related to the different ages of various parts of the colony. The theoretical results are compared to a simulation based on branching random walks. PMID- 15903468 TI - Electrostatic cancellation of gravity effects in liquid mixtures. AB - We point out that a spatially varying electric field can be used to cancel the effect of gravity in liquid mixtures by coupling to the different components' permittivities. Cancellation occurs if the system under consideration is small enough. For a simple "wedge" electrode geometry we show that the required system size and voltage are practical, and easily realizable in the laboratory. Thus this setup might be a simple alternative to other options such as the space shuttle, drop-tower, or magnetic levitation experiments. PMID- 15903469 TI - Some properties of the entropy in the natural time. AB - We show that the entropy S , defined as S identical with chi ln chi - chi ln chi [Phys. Rev. E 68, 031106 (2003)] where chi stands for the natural time [Phys. Rev. E 66, 011902 (2002)], exhibits positivity and concavity as well as stability or experimental robustness. Furthermore, the distinction between the seismic electric signal activities and "artificial" noises, based on the classification of their S values, is lost when studying the time-reversed signals. This reveals the profound importance of considering the (true) time arrow. PMID- 15903470 TI - Clustering of atoms in a model with multiple thermostats. AB - We propose a model for a one-dimensional chain of interacting particles in an external periodic potential. In this model the particles have a complex structure treated in a mean-field fashion: particle collisions are inelastic and also each particle is considered as having its own thermostat. We derived the Fokker-Planck equation for this model and demonstrated that the model has a truly equilibrium ground state. When an external dc force is applied to the atoms, the model exhibits a hysteresis even at high temperatures due to the clustering of atoms with the same velocity. Another effect of clustering is phase separation in the steady state when the system splits into regions of immobile atoms ("traffic jams") and regions of running atoms. PMID- 15903471 TI - Velocity distribution in a viscous granular gas. AB - We investigate the velocity relaxation of a viscous one-dimensional granular gas in which neither energy nor momentum is conserved in a collision. Of interest is the distribution of velocities in the gas as it cools, and the time dependence of the relaxation behavior. A Boltzmann equation of instantaneous binary collisions leads to a two-peaked distribution, as do numerical simulations of grains on a line. Of particular note is that in the presence of friction there is no inelastic collapse, so there is no need to invoke additional assumptions such as the quasielastic limit. PMID- 15903472 TI - Does electrical double layer formation lead to salt exclusion or to uptake? AB - When electric double layers are formed, cases have been reported where this formation involves expulsion of electrolyte into the solution and cases in which electrolyte is absorbed from the solution. Both situations are experimentally and theoretically documented, but they cannot be simultaneously correct. In this paper it is shown that this seeming conflict finds its cause in the way the double layer is formed: expulsion for double layers forming spontaneously by ion adsorption from the solution, but uptake when the double layer is formed by an external field. A thermodynamic analysis is presented. PMID- 15903473 TI - Dynamics of disklike clusters formed in a magnetorheological fluid under a rotational magnetic field. AB - We investigate the cluster formations and dynamics in a magnetorheological fluid under a rotational magnetic field focusing on the case of a relatively high volume fraction. We find that isotropic disklike clusters, which rotate more slowly than the field rotation, are formed at low Mason numbers (the ratio of viscous to magnetic forces) and, what is more, we show short rod clusters, which rotate stably thanks to the low Mason numbers and circulate along the surface of the disklike clusters. The circulation velocity of the surface particles is much higher than the rotational surface velocity of the rigid disklike clusters. PMID- 15903474 TI - Nematic-smectic-A phase boundary of ideally oriented Gay-Berne system: local density functional versus isothermal-isobaric Monte Carlo simulation. AB - The main focus of the present paper is on studying the nematic-smectic- A phase boundary of an ideally oriented Gay-Berne system. The phase diagram is determined by means of an isothermal-isobaric Monte Carlo simulation. The results are compared with predictions of the local density functional expanded up to second and third order in the one-particle distribution function. It is shown that generally the second-order expansion does not give satisfactory predictions for smectics. Going beyond the leading order yields good quantitative agreement at moderate densities. With increasing density the relative error of the local density functional calculations increases, but usually does not exceed 10% in densities. We conclude that the density functional approach could be competitive to time-consuming simulations in determining phase diagrams of spatially and orientationally ordered liquid crystalline structures. PMID- 15903475 TI - Comment on "test of nonequilibrium thermodynamics in glassy systems: the soft sphere case". AB - In this Comment, I show that the free energy expression for a glass proposed by La Nave [Phys. Rev. E 68, 032103 (2003)] cannot be correct since it violates the second law of thermodynamics. PMID- 15903477 TI - Set of measures to analyze the dynamics of nonequilibrium structures. AB - We present a class of statistical measures that can be used to quantify nonequilibrium surface growth. They are used to deduce information about spatiotemporal dynamics of model systems for spinodal decomposition and surface deposition. Pattern growth in the Cahn-Hilliard equation (used to model spinodal decomposition) are shown to exhibit three distinct stages. Two models of surface growth, namely, the continuous Kardar-Parisi-Zhang model and the discrete restricted-solid-on-solid model are shown to have different saturation exponents. PMID- 15903478 TI - Distance-d covering problems in scale-free networks with degree correlations. AB - A number of problems in communication systems demand the distributed allocation of network resources in order to provide better services, sampling, and distribution methods. The solution to these issues is becoming more challenging due to the increasing size and complexity of communication networks. We report here on a heuristic method to find near-optimal solutions to the covering problem in real communication networks, demonstrating that whether a centralized or a distributed design is to be used relies upon the degree correlations between connected vertices. We also show that the general belief that by targeting the hubs one can efficiently solve most problems on networks with a power-law degree distribution is not valid for assortative networks. PMID- 15903479 TI - Self-adapting network topologies in congested scenarios. AB - Most studies in complex networks assume that once a link is created between two nodes it is never deleted. However, there is a recent interest towards systems where links can be rapidly rewired. An important issue in that type of networks is to discover the topology that, given a search algorithm, optimizes the search process. In this paper, we present a system model that, depending on the current network congestion, makes nodes to establish link connections so that the resulting topologies tend to a starlike topology when congestion is small and to randomlike topologies when congestion becomes relevant. Those topologies have been shown to be optimal in the above-mentioned conditions. Such a model can be easily implemented in practice and therefore, may be relevant in areas as the topology management of peer-to-peer networks. PMID- 15903480 TI - Synchronization of non-phase-coherent chaotic electrochemical oscillations. AB - Experiments on phase and generalized synchronization of two coupled, nonidentical chaotic electrochemical oscillations are presented. We adapt measures of characterizing synchronization of a non-phase-coherent chaotic behavior and compare its properties and physicochemical mechanism to those of a phase-coherent behavior. Phase synchronization sets in along with the onset of generalized synchronization for the non-phase-coherent oscillations in contrast to phase coherent oscillations in which the phase synchronization usually occurs at a weaker coupling strength. PMID- 15903481 TI - Quantum pumping: the charge transported due to a translation of a scatterer. AB - The amount of charge that is pushed by a moving scatterer is dQ= -GdX , where dX is the displacement of the scatterer. The question is: what is G ?. Does it depend on the transmission g(0) of the scatterer? Does the answer depend on whether the system is open (with leads attached to reservoirs) or closed? In the latter case what are the implications of having "quantum chaos" and/or coupling to the environment? The answers to these questions illuminate some fundamental aspects of the theory of quantum pumping. For the analysis we take a network (graph) as a model system, and use the Kubo formula approach. PMID- 15903482 TI - Effects of symmetries on quantum fidelity decay. AB - We explore the effect of a system's symmetries on fidelity decay behavior. Chaoslike exponential fidelity decay behavior occurs in nonchaotic systems when the system possesses symmetries and the applied perturbation is not tied to a classical parameter. Similar systems without symmetries exhibit faster-than exponential decay under the same type of perturbation. This counterintuitive result, that extra symmetries cause the system to behave in a chaotic fashion, may have important ramifications for quantum-error correction. PMID- 15903483 TI - Noise-induced wave nucleations in an excitable chemical reaction. AB - We study both experimentally and numerically the temporal coherence of noise induced wave nucleations in excitable media subjected to external fluctuations with finite correlation time. The experiments are performed with the light sensitive variant of the Belousov-Zhabotinsky (BZ) reaction forced by an exponentially correlated dichotomous fluctuating illumination. We find that there exists an optimal correlation time for which nucleations coherence reaches a maximum. The same behavior is obtained in numerical simulations with a stochastic Oregonator model, modified to describe the light-sensitive BZ reaction. PMID- 15903484 TI - Simple kinetic model for fluid flows in the nanometer scale. AB - Fluid flows in the nanometer scale can be studied by molecular dynamics or Monte Carlo methods, but the time and length scales are usually limited to rather short ranges due to the computational expense. Kinetic theory is an alternative tool for studying nanoscale flows, but the existing models are rather complicated and difficult to implement. In this paper, we propose a simple Enskog-like kinetic model for nanoscale flows. The results predicted by this model compare well with molecular dynamics or Monte Carlo simulation results in the literature. PMID- 15903485 TI - Fluctuations of temperature gradients in turbulent thermal convection. AB - Broad theoretical arguments are proposed to show, formally, that the magnitude G of the temperature gradients in turbulent thermal convection at high Rayleigh numbers obeys the same advection-diffusion equation that governs the temperature fluctuation T , except that the velocity field in the new equation is substantially smoothed. This smoothed field leads to a -1 scaling of the spectrum of G in the same range of scales for which the spectral exponent of T lies between -7/5 and -5/3 . This result is confirmed by measurements in a confined container with cryogenic helium gas as the working fluid for Rayleigh number Ra=1.5x 10(11) . Also confirmed is the logarithmic form of the autocorrelation function of G . The anomalous scaling of dissipation-like quantities of T and G are identical in the inertial range, showing that the analogy between the two fields is quite deep. PMID- 15903486 TI - Direct measurement of the apparent slip length. AB - We measure velocity profiles in water flowing through thin microchannels, using particle image velocimetry combined with a nanopositioning system. From the velocity profiles, we determine the slip lengths in two cases: Smooth hydrophilic glass surfaces, and smooth hydrophobic glass surfaces, grafted with a monolayer of silane. The slip length is determined within (+/-100 nm) , i.e., five times more accurately than previous work. In all cases, we find that the slip length is below 100 nm. PMID- 15903487 TI - Majority rule dynamics in finite dimensions. AB - We investigate the long-time behavior of a majority rule opinion dynamics model in finite spatial dimensions. Each site of the system is endowed with a two-state spin variable that evolves by majority rule. In a single update event, a group of spins with a fixed (odd) size is specified and all members of the group adopt the local majority state. Repeated application of this update step leads to a coarsening mosaic of spin domains and ultimate consensus in a finite system. The approach to consensus is governed by two disparate time scales, with the longer time scale arising from realizations in which spins organize into coherent single opinion bands. The consequences of this geometrical organization on the long-time kinetics are explored. PMID- 15903488 TI - Real-time dynamics in spin-1/2 chains with adaptive time-dependent density matrix renormalization group. AB - We investigate the influence of different interaction strengths and dimerizations on the magnetization transport in antiferromagnetic spin 1/2 XXZ chains. We focus on the real-time evolution of the inhomogeneous initial state |upward arrow... upward arrow downward arrow... downward arrow > in using the adaptive time dependent density-matrix renormalization group (adaptive t-DMRG). Time scales accessible to us are of the order of 100 units of time measured in Planck's/J for almost negligible error in the observables. We find ballistic magnetization transport for small S(z) S(z) interaction and arbitrary dimerization, but almost no transport for stronger S(z) S(z) interaction, with a sharp crossover at J(z) =1 . Additionally, we perform a detailed analysis of the error made by the adaptive time-dependent DMRG using the fact that the evolution in the XX model is known exactly. We find that the error at small times is dominated by the error made by the Trotter decomposition, whereas for longer times the DMRG truncation error becomes the most important, with a very sharp crossover at some "runaway" time. Overall, errors are extremely small before the "runaway" time. PMID- 15903489 TI - Slow relaxation in the Ising model on a small-world network with strong long range interactions. AB - We consider the Ising model on a small-world network, where the long-range interaction strength J2 is in general different from the local interaction strength J1, and examine its relaxation behaviors as well as phase transitions. As J(2)/J(1) is raised from zero, the critical temperature also increases, manifesting contributions of long-range interactions to ordering. However, it becomes saturated eventually at large values of J(2)/J(1) and the system is found to display very slow relaxation, revealing that ordering dynamics is inhibited rather than facilitated by strong long-range interactions. To circumvent this problem, we propose a modified updating algorithm in Monte Carlo simulations, assisting the system to reach equilibrium quickly. PMID- 15903490 TI - Off-equilibrium generalization of the fluctuation dissipation theorem for Ising spins and measurement of the linear response function. AB - We derive for Ising spins an off-equilibrium generalization of the fluctuation dissipation theorem, which is formally identical to the one previously obtained for soft spins with Langevin dynamics [L.F. Cugliandolo, J. Kurchan, and G. Parisi, J. Phys. I 4, 1641 (1994)]. The result is quite general and holds both for dynamics with conserved and nonconserved order parameters. On the basis of this fluctuation dissipation relation, we construct an efficient numerical algorithm for the computation of the linear response function without imposing the perturbing field, which is alternative to those of Chatelain [J. Phys. A 36, 10 739 (2003)] and Ricci-Tersenghi [Phys. Rev. E 68, 065104(R) (2003)]. As applications of the new algorithm, we present very accurate data for the linear response function of the Ising chain, with conserved and nonconserved order parameter dynamics, finding that in both cases the structure is the same with a very simple physical interpretation. We also compute the integrated response function of the two-dimensional Ising model, confirming that it obeys scaling chi (t, t(w)) approximately equal to t(-a)(w) f (t/t(w)) , with a =0.26+/-0.01 , as previously found with a different method. PMID- 15903491 TI - Majority-vote model on a random lattice. AB - The stationary critical properties of the isotropic majority vote model on random lattices with quenched connectivity disorder are calculated by using Monte Carlo simulations and finite size analysis. The critical exponents gamma and beta are found to be different from those of the Ising and majority vote on the square lattice model and the critical noise parameter is found to be q(c) =0.117+/-0.005 . PMID- 15903492 TI - Equilibrium statistical mechanics of a grain boundary. AB - In this article we introduce, develop, and discuss the theoretical calculations required for the exact solution of a recently reported phase transition, the geodesic to zigzag transition. In this scenario the interfacial transition emerges from geometric competition between a geodesic, shortest path, configuration and a zigzag configuration which is able to reduce its energy by binding to a centrally positioned defect line. From a technical point of view the transition is unusual as it is described by a change from saddle dominated behavior to pole dominated behavior of the integral representing the partition function ratio. We also establish the precise fluctuation behavior of the interface by computing the spin magnetization at any point in the system. PMID- 15903493 TI - Complex networks created by aggregation. AB - We study aggregation as a mechanism for the creation of complex networks. In this evolution process vertices merge together, which increases a number of highly connected hubs. We study a range of complex network architectures produced by the aggregation. Fat-tailed (in particular, scale-free) distributions of connections are obtained for both networks with a finite number of vertices and growing networks. We observe a strong variation of a network structure with growing density of connections and find the phase transition of the condensation of edges. Finally, we demonstrate the importance of structural correlations in these networks. PMID- 15903494 TI - Geographical threshold graphs with small-world and scale-free properties. AB - Many real networks are equipped with short diameters, high clustering, and power law degree distributions. With preferential attachment and network growth, the model by Barabasi and Albert simultaneously reproduces these properties, and geographical versions of growing networks have also been analyzed. However, nongrowing networks with intrinsic vertex weights often explain these features more plausibly, since not all networks are really growing. We propose a geographical nongrowing network model with vertex weights. Edges are assumed to form when a pair of vertices are spatially close and/or have large summed weights. Our model generalizes a variety of models as well as the original nongeographical counterpart, such as the unit disk graph, the Boolean model, and the gravity model, which appear in the contexts of percolation, wire communication, mechanical and solid physics, sociology, economy, and marketing. In appropriate configurations, our model produces small-world networks with power law degree distributions. We also discuss the relation between geography, power laws in networks, and power laws in general quantities serving as vertex weights. PMID- 15903495 TI - Thermal noise limitations to force measurements with torsion pendulums: applications to the measurement of the Casimir force and its thermal correction. AB - A general analysis of thermal noise in torsion pendulums is presented. The specific case where the torsion angle is kept fixed by electronic feedback is analyzed. This analysis is applied to a recent experiment that employed a torsion pendulum to measure the Casimir force. The ultimate limit to the distance at which the Casimir force can be measured to high accuracy is discussed, and in particular we elaborate on the prospects for measuring the thermal correction. PMID- 15903496 TI - Diffuse interface approach to brittle fracture. AB - We present a continuum model for the propagation of cracks and fractures in brittle materials. The components of the strain tensor epsilon are the fundamental variables. The evolution equations are based on a free energy that reduces to that of linear elasticity for small epsilon, and accounts for cracks through energy saturation at large values of epsilon. We regularize the model by including terms dependent on gradients of epsilon in the free energy. No additional fields are introduced, and then the whole dynamics is perfectly defined. We show that the model is able to reproduce basic facts in fracture physics, like the Griffith's dependence of the critical stress as a minus one half power of the crack length. In addition, regularization makes the results insensitive to the numerical mesh used, something not at all trivial in crack modeling. We present an example of the application of the model to predict the growth and curving of cracks in a nontrivial geometrical configuration. PMID- 15903497 TI - Nonequilibrium stationary states with ratchet effect. AB - An ensemble of particles in thermal equilibrium at temperature T, modeled by Nose Hoover dynamics, moves on a triangular lattice of oriented semidisk elastic scatterers. Despite the scatterer asymmetry, a directed transport is clearly ruled out by the second law of thermodynamics. Introduction of a polarized zero mean monochromatic field creates a directed stationary flow with nontrivial dependence on temperature and field parameters. We give a theoretical estimate of directed current induced by a microwave field in an antidot superlattice in semiconductor heterostructures. PMID- 15903498 TI - Glassy behavior in an exactly solved spin system with a ferromagnetic transition. AB - We show that applying simple dynamical rules to Baxter's eight-vertex model leads to a system which resembles a glass-forming liquid. There are analogies with liquid, supercooled liquid, glassy, and crystalline states. The disordered phases exhibit strong dynamical heterogeneity at low temperatures, which may be described in terms of an emergent mobility field. Their dynamics are well described by a simple model with trivial thermodynamics, but an emergent kinetic constraint. We show that the (second order) thermodynamic transition to the ordered phase may be interpreted in terms of confinement of the excitations in the mobility field. We also describe the aging of disordered states toward the ordered phase, in terms of simple rate equations. PMID- 15903499 TI - Efficient generation of large random networks. AB - Random networks are frequently generated, for example, to investigate the effects of model parameters on network properties or to test the performance of algorithms. Recent interest in the statistics of large-scale networks sparked a growing demand for network generators that can generate large numbers of large networks quickly. We here present simple and efficient algorithms to randomly generate networks according to the most commonly used models. Their running time and space requirement is linear in the size of the network generated, and they are easily implemented. PMID- 15903500 TI - Low-temperature dynamics of kinks on Ising interfaces. AB - The anisotropic motion of an interface driven by its intrinsic curvature or by an external field is investigated in the context of the kinetic Ising model in both two and three dimensions. We derive in two dimensions (2D) a continuum evolution equation for the density of kinks by a time-dependent and nonlocal mapping to the asymmetric exclusion process. Whereas kinks execute random walks biased by the external field and pile up vertically on the physical 2D lattice, they execute hard-core biased random walks on a transformed 1D lattice. Their density obeys a nonlinear diffusion equation which can be transformed into the standard expression for the interface velocity, v=M [ (gamma+gamma'') kappa+H] , where M , gamma+gamma", and kappa are the interface mobility, stiffness, and curvature, respectively. In 3D, we obtain the velocity of a curved interface near the 100 orientation from an analysis of the self-similar evolution of 2D shrinking terraces. We show that this velocity is consistent with the one predicted from the 3D tensorial generalization of the law for anisotropic curvature-driven motion. In this generalization, both the interface stiffness tensor and the curvature tensor are singular at the 100 orientation. However, their product, which determines the interface velocity, is smooth. In addition, we illustrate how this kink-based kinetic description provides a useful framework for studying more complex situations by modeling the effect of immobile dilute impurities. PMID- 15903501 TI - Constrained tricritical phenomena in two dimensions. AB - We investigate several tricritical models on the square lattice by means of Monte Carlo simulations. These include the Blume-Capel model, Baxter's hard-square model, and the q=1 , 3, and 4 Potts models with vacancies. We use a combination of the Wolff and geometric cluster methods, which conserves the total number of vacancies or lattice-gas particles and suppresses critical slowing down. Several quantities are sampled, such as the specific heat C and the structure factor C(s) , which accounts for the large-scale spatial inhomogeneity of the energy fluctuations. We find that the constraint strongly modifies some of the critical singularities. For instance, the specific heat C reaches a finite value at tricriticality, while C(s) remains divergent as in the unconstrained system. We are able to explain these observed constrained phenomena on the basis of the Fisher renormalization mechanism generalized to include a subleading relevant thermal scaling field. In this context, we find that, under the constraint, the leading thermal exponent y(t1) is renormalized to 2- y(t1) , while the subleading exponent y(t2) remains unchanged. PMID- 15903502 TI - Analysis of ensemble learning using simple perceptrons based on online learning theory. AB - Ensemble learning of K nonlinear perceptrons, which determine their outputs by sign functions, is discussed within the framework of online learning and statistical mechanics. One purpose of statistical learning theory is to theoretically obtain the generalization error. This paper shows that ensemble generalization error can be calculated by using two order parameters, that is, the similarity between a teacher and a student, and the similarity among students. The differential equations that describe the dynamical behaviors of these order parameters are derived in the case of general learning rules. The concrete forms of these differential equations are derived analytically in the cases of three well-known rules: Hebbian learning, perceptron learning, and AdaTron (adaptive perceptron) learning. Ensemble generalization errors of these three rules are calculated by using the results determined by solving their differential equations. As a result, these three rules show different characteristics in their affinity for ensemble learning, that is "maintaining variety among students." Results show that AdaTron learning is superior to the other two rules with respect to that affinity. PMID- 15903503 TI - Level density for deformations of the Gaussian orthogonal ensemble. AB - Formulas are derived for the average level density of deformed, or transition, Gaussian orthogonal random matrix ensembles. After some general considerations about Gaussian ensembles, we derive formulas for the average level density for (i) the transition from the Gaussian orthogonal ensemble (GOE) to the Poisson ensemble and (ii) the transition from the GOE to m GOEs. PMID- 15903504 TI - Network growth by copying. AB - We introduce a growing network model in which a new node attaches to a randomly selected node, as well as to all ancestors of the target node. This mechanism produces a sparse, ultrasmall network where the average node degree grows logarithmically with network size while the network diameter equals 2. We determine basic geometrical network properties, such as the size dependence of the number of links and the in- and out-degree distributions. We also compare our predictions with real networks where the node degree also grows slowly with time- the Internet and the citation network of all Physical Review papers. PMID- 15903505 TI - Memory effects on the statistics of fragmentation. AB - We investigate through extensive molecular dynamics simulations the fragmentation process of two-dimensional Lennard-Jones systems. After thermalization, the fragmentation is initiated by a sudden increment to the radial component of the particles' velocities. We study the effect of temperature of the thermalized system as well as the influence of the impact energy of the "explosion" event on the statistics of mass fragments. Our results indicate that the cumulative distribution of fragments follows the scaling ansatz F(m) proportional to m(1 alpha)exp-(m/m(0))(gamma), where m is the mass, m(0) and gamma are cutoff parameters, and alpha is a scaling exponent that is dependent on the temperature. More precisely, we show clear evidence that there is a characteristic scaling exponent alpha for each macroscopic phase of the thermalized system, i.e., that the nonuniversal behavior of the fragmentation process is dictated by the state of the system before it breaks down. PMID- 15903506 TI - Exact probability function for bulk density and current in the asymmetric exclusion process. AB - We examine the asymmetric simple exclusion process with open boundaries, a paradigm of driven diffusive systems, having a nonequilibrium steady-state transition. We provide a full derivation and expanded discussion and digression on results previously reported briefly in M. Depken and R. Stinchcombe, Phys. Rev. Lett. 93, 040602 (2004). In particular we derive an exact form for the joint probability function for the bulk density and current, both for finite systems, and also in the thermodynamic limit. The resulting distribution is non-Gaussian, and while the fluctuations in the current are continuous at the continuous phase transitions, the density fluctuations are discontinuous. The derivations are done by using the standard operator algebraic techniques and by introducing a modified version of the original operator algebra. As a by-product of these considerations we also arrive at a very simple way of calculating the normalization constant appearing in the standard treatment with the operator algebra. Like the partition function in equilibrium systems, this normalization constant is shown to completely characterize the fluctuations, albeit in a very different manner. PMID- 15903507 TI - Instability of pedestrian flow and phase structure in a two-dimensional optimal velocity model. AB - A two-dimensional optimal velocity model was proposed for the study of pedestrian and granular flow. We investigate the stability of homogeneous flow in the linear approximation and show the phase diagram of the model. We also investigate the property of the model by numerical simulation in the cases of unidirectional and counter flow. From these results, we present a unified understanding of the behavior of pedestrians and other related systems. PMID- 15903508 TI - Efficiency of informational transfer in regular and complex networks. AB - We analyze the process of informational exchange through complex networks by measuring network efficiencies. Aiming to study nonclustered systems, we propose a modification of this measure on the local level. We apply this method to an extension of the class of small worlds that includes declustered networks and show that they are locally quite efficient, although their clustering coefficient is practically zero. Unweighted systems with small-world and scale-free topologies are shown to be both globally and locally efficient. Our method is also applied to characterize weighted networks. In particular we examine the properties of underground transportation systems of Madrid and Barcelona and reinterpret the results obtained for the Boston subway network. PMID- 15903509 TI - Crack propagation in viscoelastic solids. AB - We study crack propagation in a viscoelastic solid. Using simple arguments, we derive equations for the velocity dependence of the crack-tip radius, a (v) , and for the energy per unit area to propagate the crack, G (v) . For a viscoelastic modulus E (omega) which increases as omega(1-s) (0< s< 1) in the transition region between the rubbery region and the glassy region, we find that a (v) approximately G (v) approximately v(alpha) with alpha= (1-s) / (2-s) . The theory is in good agreement with experiment. PMID- 15903510 TI - Statistics of weighted treelike networks. AB - We study the statistics of growing networks with a tree topology in which each link carries a weight (k(i) k(j))(theta) , where k(i) and k(j) are the node degrees at the end points of link ij . Network growth is governed by preferential attachment in which a newly added node attaches to a node of degree k with rate A(k) =k+lambda . For general values of theta and lambda , we compute the total weight of a network as a function of the number of nodes N and the distribution of link weights. Generically, the total weight grows as N for lambda>theta-1 and superlinearly otherwise. The link weight distribution is predicted to have a power-law form that is modified by a logarithmic correction for the case lambda=0 . We also determine the node strength, defined as the sum of the weights of the links that attach to the node, as function of k . Using known results for degree correlations, we deduce the scaling of the node strength on k and N . PMID- 15903511 TI - Stochastic resonance in bulk semiconductor lasers. AB - The stochastic time scale of the mode hopping in a bulk semiconductor laser can be varied maintaining the symmetry of the residence times by a proper tuning of the laser substrate temperature and pumping current. While the addition of external noise to the pumping current affects the symmetry of the mode-hopping process, a sinusoidal modulation does not, providing that the modulation amplitude is below a critical value. In this case, we observe stochastic resonance in the modal intensities of the laser. We show the occurrence of the phenomenon in the spectral domain, and we characterize it by a statistical analysis based on the residence times probability distributions. The evidence of bona fide resonance is also provided, varying the modulation frequency and analyzing a proper statistical indicator. Changing the temperature of the laser substrate we show that resonance occurs at different modulation periods always equal to the double of the average residence time measured without modulation. PMID- 15903512 TI - Work distribution functions in polymer stretching experiments. AB - We compute the distribution of the work done in stretching a Gaussian polymer, made of N monomers, at a finite rate. For a one-dimensional polymer undergoing Rouse dynamics, the work distribution is a Gaussian and we explicitly compute the mean and width. The two cases where the polymer is stretched, either by constraining its end or by constraining the force on it, are examined. We discuss connections to Jarzynski's equality and the fluctuation theorems. PMID- 15903513 TI - Rate equation approach for correlations in growing network models. AB - We propose a rate equation approach to compute two vertex correlations in scale free growing network models based on the preferential attachment mechanism. The formalism, based on previous work of Szabo [Phys. Rev. E. 67, 056102 (2002)] for the clustering spectrum, measuring three vertex correlations, is based on a rate equation in the continuous degree and time approximation for the average degree of the nearest neighbors of vertices of degree k , with an appropriate boundary condition. We study the properties of both two and three vertex correlations for linear preferential attachment models, and also for a model yielding a large clustering coefficient. The expressions obtained are checked by means of extensive numerical simulations. The rate equation proposed can be generalized to more sophisticated growing network models, and also extended to deal with related correlation measures. As an example, we consider the case of a recently proposed model of weighted networks, for which we are able to compute a weighted two vertex correlation function, taking into account the strength of the interactions between connected vertices. PMID- 15903514 TI - Spacetime structures of continuous-time quantum walks. AB - The propagation by continuous-time quantum walks (CTQWs) on one-dimensional lattices shows structures in the transition probabilities between different sites reminiscent of quantum carpets. For a system with periodic boundary conditions, we calculate the transition probabilities for a CTQW by diagonalizing the transfer matrix and by a Bloch function ansatz. Remarkably, the results obtained for the Bloch function ansatz can be related to results from (discrete) generalized coined quantum walks. Furthermore, we show that here the first revival time turns out to be larger than for quantum carpets. PMID- 15903515 TI - Persistence of randomly coupled fluctuating interfaces. AB - We study the persistence properties in a simple model of two coupled interfaces characterized by heights h(1) and h(2) , respectively, each growing over a d dimensional substrate. The first interface evolves independently of the second and can correspond to any generic growing interface, e.g., of the Edwards Wilkinson or of the Kardar-Parisi-Zhang variety. The evolution of h(2) , however, is coupled to h(1) via a quenched random velocity field. In the limit d-->0 , our model reduces to the Matheron-de Marsily model in two dimensions. For d=1, our model describes a Rouse polymer chain in two dimensions advected by a transverse velocity field. We show analytically that after a long waiting time t(0) - >infinity , the stochastic process h(2) , at a fixed point in space but as a function of time, becomes a fractional Brownian motion with a Hurst exponent, H2 =1- beta(1) /2 , where beta(1) is the growth exponent characterizing the first interface. The associated persistence exponent is shown to be theta(2)(s) =1- H2 = beta(1) /2 . These analytical results are verified by numerical simulations. PMID- 15903516 TI - Finite-size scaling and universality for the totally asymmetric simple-exclusion process. AB - The applicability of the concepts of finite-size scaling and universality to nonequilibrium phase transitions is considered in the framework of the one dimensional totally asymmetric simple-exclusion process with open boundaries. In the thermodynamic limit there are boundary-induced transitions both of the first and second order between steady-state phases of the model. We derive finite-size scaling expressions for the current near the continuous phase transition and for the local density near the first-order transition under different stochastic dynamics and compare them to establish the existence of universal functions. Next we study numerically the finite-size behavior of the Lee-Yang zeros of the normalization factor for the different steady-state probabilities. PMID- 15903517 TI - Growing network model for community with group structure. AB - We propose a growing network model for a community with a group structure. The community consists of individual members and groups, gatherings of members. The community grows as a new member is introduced by an existing member at each time step. The new member then creates a new group or joins one of the groups of the introducer. We investigate the emerging community structure analytically and numerically. The group size distribution shows a power-law distribution for a variety of growth rules, while the activity distribution follows an exponential or a power law depending on the details of the growth rule. We also present an analysis of empirical data from online communities the "Groups" in http://www.yahoo.com and the "Cafe" in http://www.daum.net, which show a power law distribution for a wide range of group sizes. PMID- 15903518 TI - Flexible construction of hierarchical scale-free networks with general exponent. AB - Extensive studies have been done to understand the principles behind architectures of real networks. Recently, evidence for hierarchical organization in many real networks has also been reported. Here, we present a hierarchical model that reproduces the main experimental properties observed in real networks: scale-free of degree distribution P (k) [frequency of the nodes that are connected to k other nodes decays as a power law P (k) approximately k(-gamma) ] and power-law scaling of the clustering coefficient C (k) approximately k(-1) . The major points of our model can be summarized as follows. (a) The model generates networks with scale-free distribution for the degree of nodes with general exponent gamma>2 , and arbitrarily close to any specified value, being able to reproduce most of the observed hierarchical scale-free topologies. In contrast, previous models cannot obtain values of gamma>2.58 . (b) Our model has structural flexibility because (i) it can incorporate various types of basic building blocks (e.g., triangles, tetrahedrons, and, in general, fully connected clusters of n nodes) and (ii) it allows a large variety of configurations (i.e., the model can use more than n-1 copies of basic blocks of n nodes). The structural features of our proposed model might lead to a better understanding of architectures of biological and nonbiological networks. PMID- 15903519 TI - Evolutionary prisoner's dilemma game on hierarchical lattices. AB - An evolutionary prisoner's dilemma (PD) game is studied with players located on a hierarchical structure of layered square lattices. The players can follow two strategies [ D (defector) and C (cooperator)] and their income comes from PD games with the "neighbors." The adoption of one of the neighboring strategies is allowed with a probability dependent on the payoff difference. Monte Carlo simulations are performed to study how the measure of cooperation is affected by the number of hierarchical levels (Q) and by the temptation to defect. According to the simulations the highest frequency of cooperation can be observed at the top level if the number of hierarchical levels is low (Q<4) . For larger Q , however, the highest frequency of cooperators occurs in the middle layers. The four-level hierarchical structure provides the highest average (total) income for the whole community. PMID- 15903520 TI - Noise-enhanced categorization in a recurrently reconnected neural network. AB - We investigate the interplay of recurrence and noise in neural networks trained to categorize spatial patterns of neural activity. We develop the following procedure to demonstrate how, in the presence of noise, the introduction of recurrence permits to significantly extend and homogenize the operating range of a feed-forward neural network. We first train a two-level perceptron in the absence of noise. Following training, we identify the input and output units of the feed-forward network, and thus convert it into a two-layer recurrent network. We show that the performance of the reconnected network has features reminiscent of nondynamic stochastic resonance: the addition of noise enables the network to correctly categorize stimuli of subthreshold strength, with optimal noise magnitude significantly exceeding the stimulus strength. We characterize the dynamics leading to this effect and contrast it to the behavior of a more simple associative memory network in which noise-mediated categorization fails. PMID- 15903521 TI - Statistical theory of Internet exploration. AB - The general methodology used to construct Internet maps consists in merging all the discovered paths obtained by sending data packets from a set of active computers to a set of destination hosts, obtaining a graphlike representation of the network. This technique, sometimes referred to as Internet tomography, spurs the issue concerning the statistical reliability of such empirical maps. We tackle this problem by modeling the network sampling process on synthetic graphs and by using a mean-field approximation to obtain expressions for the probability of edge and vertex detection in the sampled graph. This allows a general understanding of the origin of possible sampling biases. In particular, we find a direct dependence of the map statistical accuracy upon the topological properties (in particular, the betweenness centrality property) of the underlying network. In this framework, it appears that statistically heterogeneous network topologies are captured better than the homogeneous ones during the mapping process. Finally, the analytical discussion is complemented with a thorough numerical investigation of simulated mapping strategies in network models with varying topological properties. PMID- 15903522 TI - Possible crossover of a nonuniversal quantity at the upper critical dimension. AB - We report on a possible crossover of a nonuniversal quantity at the upper critical dimensionality in the field of percolation. Plotting recent estimates for site percolation thresholds of hypercubes in dimension 6 < or =d < or =13 against corresponding predictions from the Galam-Mauger (GM) formula p(c) = p(0) [(d-1) (q-1)](-a) d(b) for percolation thresholds, a significant departure of p(c) is observed for d > or =6 . This result is reminiscent of the crossover undergone by universal quantities in critical phenomena. For bond percolation, evidence of such a crossover of dimensionality would require an improvement of the GM formula to reach a relative error of typically 0.2%, while it is currently at 0.9% for hypercubes. PMID- 15903523 TI - Nontrivial temporal scaling in a Galilean stick-slip dynamics. AB - We examine the stick-slip fluctuating response of a rough massive nonrotating cylinder moving on a rough inclined groove which is submitted to weak external perturbations and which is maintained well below the angle of repose. The experiments presented here, which are reminiscent of Galileo's works with rolling objects on inclines, have brought in the last years important insights into the friction between surfaces in relative motion and are of relevance for earthquakes, differing from classical block-spring models by the mechanism of energy input in the system. Robust nontrivial temporal scaling laws appearing in the dynamics of this system are reported, and it is shown that the time-support where dissipation occurs approaches a statistical fractal set with a fixed value of dimension. The distribution of periods of inactivity in the intermittent motion of the cylinder is also studied and found to be closely related to the lacunarity of a random version of the classic triadic Cantor set on the line. PMID- 15903524 TI - Monte Carlo study of hard pentagons. AB - How does a liquid freeze if the geometry of its particles conflicts with the symmetry of the crystal it should naturally form? We study this question in the simplest model system of particles exhibiting such a symmetry mismatch: hard pentagons in two dimensions. Using isobaric and isotensic Monte Carlo simulations we have studied the phase behavior of hard pentagons. On increasing the pressure from the homogeneous and isotropic low-density phase, the system first exhibits a rotator phase (plastic solid) with a triangular lattice structure. At higher densities it undergoes a weak first-order phase transition into a "striped" phase composed of alternating rows of oppositely pointing particles. This phase is analogous to the "striped" phase in the compressible antiferromagnetic Ising model on a triangular lattice and is an example of systems in which frustration due to the mismatch in symmetries is released by an elastic coupling to the lattice. In order to pursue this analogy we also consider hard heptagons, showing that in this case the decrease in symmetry mismatch indeed leads to a shift in the transition densities to higher values and a weakening of the transition. PMID- 15903525 TI - Critical behavior of an Ising system on the Sierpinski carpet: a short-time dynamics study. AB - The short-time dynamic evolution of an Ising model embedded in an infinitely ramified fractal structure with noninteger Hausdorff dimension was studied using Monte Carlo simulations. Completely ordered and disordered spin configurations were used as initial states for the dynamic simulations. In both cases, the evolution of the physical observables follows a power-law behavior. Based on this fact, the complete set of critical exponents characteristic of a second-order phase transition was evaluated. Also, the dynamic exponent theta of the critical initial increase in magnetization, as well as the critical temperature, were computed. The exponent theta exhibits a weak dependence on the initial (small) magnetization. On the other hand, the dynamic exponent z shows a systematic decrease when the segmentation step is increased, i.e., when the system size becomes larger. Our results suggest that the effective noninteger dimension for the second-order phase transition is noticeably smaller than the Hausdorff dimension. Even when the behavior of the magnetization (in the case of the ordered initial state) and the autocorrelation (in the case of the disordered initial state) with time are very well fitted by power laws, the precision of our simulations allows us to detect the presence of a soft oscillation of the same type in both magnitudes that we attribute to the topological details of the generating cell at any scale. PMID- 15903526 TI - Markov chain-based numerical method for degree distributions of growing networks. AB - In this paper, we establish a relation between growing networks and Markov chains, and propose a computational approach for network degree distributions. Using the Barabasi-Albert model as an example, we first show that the degree evolution of a node in a growing network follows a nonhomogeneous Markov chain. Exploring the special structure of these Markov chains, we develop an efficient algorithm to compute the degree distribution numerically with a computation complexity of O (t(2)), where t is the number of time steps. We use three examples to demonstrate the computation procedure and compare the results with those from existing methods. PMID- 15903527 TI - Extension of scaled particle theory to inhomogeneous hard particle fluids. I. Cavity growth at a hard wall. AB - The methods of traditional scaled particle theory (SPT) are used to develop an extended scaled particle theory that is now applicable to hard particle fluids confined by hard walls. The new theory, labeled inhomogeneous SPT (I-SPT), introduces the function G that describes the average value of the anisotropic density of hard particle centers contacting a cavity placed at or behind a hard wall. We present an exact relation describing G for certain cavity sizes and, similar to the original SPT, an accurate interpolation scheme for larger cavity radii. Given G , the reversible work of inserting a cavity centered at or behind the hard wall can be estimated. The work predictions at low to moderate packing fractions are extremely accurate, though small deviations from simulation results become apparent at packing fractions close to the bulk hard sphere freezing transition. I-SPT also reveals the importance of the line tension in determining the free energy of cavity formation for cavities intersecting a hard wall, a term which has been previously neglected. Furthermore, this paper provides the initial groundwork needed to develop a more complete SPT-based theory that can accurately predict the depletion force between a hard particle and a hard structureless wall. PMID- 15903528 TI - Extension of scaled particle theory to inhomogeneous hard particle fluids. II. Theory and simulation of fluid structure surrounding a cavity that intersects a hard wall. AB - Integral equations describing the structure of a hard sphere fluid surrounding a cavity that intersects a hard wall are derived from scaled particle theory (SPT). The new expressions are solved exactly for specific cavity radii and the predictions are compared to simulation-generated results, showing excellent agreement. Additional simulation studies are conducted for cavity radii that fall outside the range of exact solution. For all cavity sizes, an enhancement of the local density of hard spheres over that of the hard wall contact value is seen for positions near the point of intersection of the cavity and the hard wall. The local density in front of the cavity and away from the hard wall is depleted at small cavity sizes, but eventually approaches the density profile created by a cavity placed within a bulk hard sphere fluid at larger cavity radii. The exact solutions and simulation results are also used to understand why a minimum appears in the inhomogeneous SPT function G (lambda,h) [D. W. Siderius and D. S. Corti, preceding paper, Phys. Rev. E 71, 036141 (2005)]. PMID- 15903529 TI - Criteria for evaluation of universal formulas for percolation thresholds. AB - Several universal formulas that predict approximate values for percolation thresholds of all periodic graphs have been proposed in the physics and engineering literature. The existing universal formulas have been found to have substantial errors in their predictions for some lattices. This paper proposes a set of desirable criteria for universal formulas to satisfy, and investigates which criteria are satisfied by two bond threshold formulas and two site threshold formulas most cited in the literature. The analysis is limited to lattices in two dimensions. PMID- 15903530 TI - Exactly solvable scale-free network model. AB - We study a deterministic scale-free network recently proposed by Barabasi, Ravasz, and Vicsek. We find that there are two types of nodes: the hub and rim nodes, which form a bipartite structure of the network. We first derive the exact numbers P (k) of nodes with degree k for the hub and rim nodes in each generation of the network, respectively. Using this, we obtain the exact exponents of the distribution function P (k) of nodes with k degree in the asymptotic limit of k- >infinity . We show that the degree distribution for the hub nodes exhibits the scale-free nature, P (k) proportional to k(-gamma) with gamma=ln 3/ln 2=1.584 962 , while the degree distribution for the rim nodes is given by P(k) proportional to e(-gamma'k) with gamma' =ln (3/2) =0.405 465 . Second, we analytically calculate the second-order average degree of nodes, d(-) . Third, we numerically as well as analytically calculate the spectra of the adjacency matrix A for representing topology of the network. We also analytically obtain the exact number of degeneracies at each eigenvalue in the network. The density of states (i.e., the distribution function of eigenvalues) exhibits the fractal nature with respect to the degeneracy. Fourth, we study the mathematical structure of the determinant of the eigenequation for the adjacency matrix. Fifth, we study hidden symmetry, zero modes, and its index theorem in the deterministic scale-free network. Finally, we study the nature of the maximum eigenvalue in the spectrum of the deterministic scale-free network. We will prove several theorems for it, using some mathematical theorems. Thus, we show that most of all important quantities in the network theory can be analytically obtained in the deterministic scale-free network model of Barabasi, Ravasz, and Vicsek. Therefore, we may call this network model the exactly solvable scale-free network. PMID- 15903531 TI - Shear thinning of a critical viscoelastic fluid. AB - The frequency and shear dependent critical viscosity at a correlation length xi= kappa(-1) has the form eta= eta(0) kappa(- x(eta) ) G ( z(1) , z(2) ) , where z(1) and z(2) are the independent dimensionless numbers in the problem defined as z(1) =-iomega/2 Gamma(0) kappa(3) and z(2) =-iomega/2 Gamma(0) kappa(3)(c) . The decay rate of critical fluctuations of correlation length kappa(-1) is Gamma(0) kappa(3) and k(c) is the effective wave number for which Gamma(0) k(3)(c) =S , the shear rate. The function G ( z(1) , z(2) ) is calculated in a one-loop self consistent theory. PMID- 15903532 TI - Spin glasses in the limit of an infinite number of spin components. AB - We consider spin glass models in which the number of spin components m is infinite. In the formulation of the problem appropriate for numerical calculations proposed by several authors, we show that the order parameter defined by the long-distance limit of the correlation functions is actually zero and there is only "quasi-long-range order" below the transition temperature. Nonetheless, there can be a finite temperature phase transition where the decay of correlations changes from exponential to power law. We also show that the spin glass transition temperature is zero in three dimensions so power-law behavior only occurs at T=0 in this case. We also argue that the order of limits, m- >infinity and N-->infinity is important, where N is the number of spins. PMID- 15903533 TI - Entropy production in diffusion-reaction systems: the reactive random Lorentz gas. AB - We report the study of a random Lorentz gas with a reaction of isomerization A<==>B between two colors of moving particles elastically bouncing on hard disks. The reaction occurs when the moving particles collide on catalytic disks, which constitute a fraction of all the disks. Under dilute-gas conditions, the reaction diffusion process is ruled by two coupled Boltzmann-Lorentz equations for the distribution functions of the colors. The macroscopic reaction-diffusion equations with cross-diffusion terms induced by the chemical reaction are derived from the kinetic equations. We use an H theorem of the kinetic theory in order to derive a macroscopic entropy depending on the gradients of color densities and which has a non-negative entropy production in agreement with the second law of thermodynamics. PMID- 15903534 TI - Quasistationary trajectories of the mean-field XY Hamiltonian model: a topological perspective. AB - We employ a topological approach to investigate the nature of quasistationary states of the mean-field XY Hamiltonian model. We focus on the quasistationary states reached when the system is initially prepared in a fully magnetized configuration. By means of numerical simulations and analytical considerations, we show that, along the quasistationary trajectories, the system evolves in a manifold of critical points of the potential energy function. Although these critical points are maxima, the large number of directions with marginal stability may be responsible for the slow relaxation dynamics and the trapping of the system in such trajectories. PMID- 15903535 TI - Crossover behavior in a mixed-mode fiber bundle model. AB - We introduce a mixed-mode load sharing scheme in a fiber bundle model. This model reduces exactly to equal-load-sharing (ELS) and local-load-sharing (LLS) models at the two extreme limits of a single-load-sharing parameter. We identify two distinct regimes: (a) the mean-field regime where the ELS mode dominates and (b) the short-range regime dominated by the LLS mode. The crossover behavior is explored through a numerical study of the strength variation, the avalanche statistics, susceptibility and relaxation time variations, the correlations among the broken fibers, and their cluster analysis. Analyzing the moments of the cluster size distributions we locate the crossover point of these regimes. We thus conclude that even in one dimension, the fiber bundle model shows crossover behavior from mean-field to short-range interactions. PMID- 15903536 TI - Cooperative dynamics in a network of stochastic elements with delayed feedback. AB - Networks of globally coupled, noise-activated, bistable elements with connection time delays are considered. The dynamics of these systems is studied numerically using a Langevin description and analytically using (1) a Gaussian approximation as well as (2) a dichotomous model. The system demonstrates ordering phase transitions and multistability. That is, for a strong enough feedback it exhibits nontrivial stationary states and oscillatory states whose frequencies depend only on the mean of the time delay distribution function. Other observed dynamical phenomena include coherence resonance and, in the case of nonuniform coupling strengths, amplitude death and chaos. Furthermore, an increase of the stability of the trivial equilibrium with increasing nonuniformity of the time delays is observed. PMID- 15903537 TI - Onset of synchronization in large networks of coupled oscillators. AB - We study the transition from incoherence to coherence in large networks of coupled phase oscillators. We present various approximations that describe the behavior of an appropriately defined order parameter past the transition and generalize recent results for the critical coupling strength. We find that, under appropriate conditions, the coupling strength at which the transition occurs is determined by the largest eigenvalue of the adjacency matrix. We show how, with an additional assumption, a mean-field approximation recently proposed is recovered from our results. We test our theory with numerical simulations and find that it describes the transition when our assumptions are satisfied. We find that our theory describes the transition well in situations in which the mean field approximation fails. We study the finite-size effects caused by nodes with small degree and find that they cause the critical coupling strength to increase. PMID- 15903538 TI - Topology and phase transitions: from an exactly solvable model to a relation between topology and thermodynamics. AB - The elsewhere surmized topological origin of phase transitions is given here important evidence through the analytic study of an exactly solvable model for which both topology of submanifolds of configuration space and thermodynamics are worked out. The model is a mean-field one with a k-body interaction. It undergoes a second-order phase transition for k=2 and a first-order one for k >2 . This opens a perspective for the understanding of the deep origin of first and second order phase transitions, respectively. In particular, a remarkable theoretical result consists of a mathematical characterization of first-order transitions. Moreover, we show that a "reduced" configuration space can be defined in terms of collective variables, such that the correspondence between phase transitions and topology changes becomes one-to-one, for this model. Finally, an unusual relationship is worked out between the microscopic description of a classical N body system and its macroscopic thermodynamic behavior. This consists of a functional dependence of thermodynamic entropy upon the Morse indexes of the critical points (saddles) of the constant energy hypersurfaces of the microscopic 2N-dimensional phase space. Thus phase space (and configuration space) topology is directly related to thermodynamics. PMID- 15903539 TI - Four height variables, boundary correlations, and dissipative defects in the Abelian sandpile model. AB - We analyze the two-dimensional Abelian sandpile model, and demonstrate that the four height variables have different field identifications in the bulk, and along closed boundaries, but become identical, up to rescaling, along open boundaries. We consider two-point boundary correlations in detail, and discuss a number of complications that arise in the mapping from sandpile correlations to spanning tree correlations; the structure of our results suggests a conjecture that could greatly simplify future calculations. We find a number of three-point functions along closed boundaries, and propose closed boundary field identifications for the height variables. We analyze the effects of dissipative defect sites, at which the number of grains is not conserved, and show that dissipative defects along closed boundaries, and in the bulk, have no effect on any weakly allowed cluster variables, or on their correlations. Along open boundaries, we find a particularly simple field structure; we calculate all n-point correlations, for any combinations of height variables and dissipative defect sites, and find that all heights and defects are represented by the same field operator. PMID- 15903540 TI - Effect of spatial reflection symmetry on the distribution of the parametric conductance derivative in ballistic chaotic cavities. AB - We study the effect of left-right symmetry on the distribution of the parametric derivative of the dimensionless conductance T with respect to an external parameter X , partial differentialT/ partial differentialX , of ballistic chaotic cavities with two leads, each supporting N propagating modes. We show that T and partial differentialT/ partial differentialX are linearly uncorrelated for any N . For N=1 we calculate the distribution of partial differentialT/ partial differentialX in the presence and absence of time-reversal invariance. In both cases, it has a logarithmic singularity at zero derivative and algebraic tails with an exponent different from the one of the asymmetric case. We also obtain explicit analytical results for the mean and variance of the distribution of partial differentialT/ partial differentialX for arbitrary N . Numerical simulations are performed for N=5 and 10 to show that the distribution P ( partial differentialT/ partial differentialX) tends towards a Gaussian one when N increases. PMID- 15903541 TI - Mechanisms for dynamic crack branching in brittle elastic solids: strain field kinematics and reflected surface waves. AB - We report on a numerical simulation study of dynamic fracture in strip-shaped plates in which we implement a fracture criterion that fully respects mode-I symmetry. The crack dynamics is studied as a function of the length of the initial notch. The cracks show accelerated straight motion until branching appears. We show that branching can be triggered by two different mechanisms: namely, the kinematics of the strain field and back-reflected surface waves traveling on the crack lip. We also propose a qualitative explanation for the kinematic branching mechanism in terms of the effects of the lattice discretization on Yoffe's stress field. The kinematic branching mechanism is understood by analyzing the disconnection times of the nodes ahead and aside of the crack tip. PMID- 15903542 TI - Mode expansion description of stadium-cavity laser dynamics. AB - The lasing dynamics of a stadium-cavity laser is studied by using a mode expansion model which is a reduction of the Schrodinger-Bloch model. We study the properties of stationary lasing states when two cavity modes are selectively excited, while examining the validity of the mode expansion model by comparing its results with those of the Schrodinger-Bloch model. Some analytical results are obtained for single-mode and two-mode stationary lasing states for the mode expansion model. PMID- 15903543 TI - Nonlinear motion of optically torqued nanorods. AB - We apply light torques to single optically trapped glass nanorods suspended in water. The resulting motion is studied experimentally and consists of two distinct regimes: a linear regime where the rod angle increases linearly with time and a nonlinear regime where the rod angle changes nonlinearly, experiencing accelerations and rapid reversals. We present a detailed theoretical treatment for the motion of such nanorods, which agrees extremely well with the observed motion. The experiments are carried out so that the trapped and torqued nanorods move without influence from surfaces. Such a model system is critical to understanding the more complex motion that occurs near a surface. Studying such nonlinear motion both free of, and near, a surface is important for understanding nanofluidics and hydrodynamic motion at the nanoscale. PMID- 15903544 TI - Enhanced tracer transport by the spiral defect chaos state of a convecting fluid. AB - To understand how spatiotemporal chaos may modify material transport, we use direct numerical simulations of the three-dimensional Boussinesq equations and of an advection-diffusion equation to study the transport of a passive tracer by the spiral defect chaos state of a convecting fluid. The simulations show that the transport is diffusive and is enhanced by the spatiotemporal chaos. The enhancement in tracer diffusivity follows two regimes. For large Peclet numbers (that is, small molecular diffusivities of the tracer), we find that the enhancement is proportional to the Peclet number. For small Peclet numbers, the enhancement is proportional to the square root of the Peclet number. We explain the presence of these two regimes in terms of how the local transport depends on the local wave numbers of the convection rolls. For large Peclet numbers, we further find that defects cause the tracer diffusivity to be enhanced locally in the direction orthogonal to the local wave vector but suppressed in the direction of the local wave vector. PMID- 15903545 TI - Excitation of multiphase waves of the nonlinear Schrodinger equation by capture into resonances. AB - A method for adiabatic excitation and control of multiphase ( N -band) waves of the periodic nonlinear Schrodinger (NLS) equation is developed. The approach is based on capturing the system into successive resonances with external, small amplitude plane waves having slowly varying frequencies. The excitation proceeds from zero and develops in stages, as an (N+1) -band (N=0,1,2,...) , growing amplitude wave is formed in the (N+1) th stage from an N -band solution excited in the preceding stage. The method is illustrated in simulations, where the excited multiphase waves are analyzed via the spectral approach of the inverse scattering transform method. The theory of excitation of 0- and 1-band NLS solutions by capture into resonances is developed on the basis of a weakly nonlinear version of Whitham's averaged variational principle. The phenomenon of thresholds on the driving amplitudes for capture into successive resonances and the stability of driven, phase-locked solutions in these cases are discussed. PMID- 15903546 TI - Detection of weak directional coupling: phase-dynamics approach versus state space approach. AB - We compare two conceptually different approaches to the detection of weak directional couplings between two oscillatory systems from bivariate time series. The first approach is based on the analysis of the systems' phase dynamics, whereas the other one tests for interdependencies in the reconstructed state spaces of the systems. We analyze the sensitivity of both techniques to weak couplings in numerical experiments by considering couplings between almost identical as well as between significantly different nonlinear systems. We study different degrees of phase diffusion, test the robustness of the two techniques against observational noise, and investigate the influence of the time series length. Our results show that none of the two approaches is generally superior to the other, and we conclude that it is probably the combination of both techniques that would allow the most comprehensive and reliable characterization of coupled systems. PMID- 15903547 TI - Analysis of multiple time scales in a transistor amplifier. AB - It was shown previously in an experiment that when high frequency signals (on the order of 1 MHz) were injected into this low frequency amplifier, the nonlinearities of the pn junctions caused period doubling, chaos, and very low frequency oscillations (on the order of 1 Hz). In this paper we present theory and simulations to explain the existence of the low frequency oscillations. PMID- 15903548 TI - Bistable chaos without symmetry in generalized synchronization. AB - Frequently, multistable chaos is found in dynamical systems with symmetry. We demonstrate a rare example of bistable chaos in generalized synchronization (GS) in coupled chaotic systems without symmetry. Bistable chaos in GS refers to two chaotic attractors in the response system which both synchronize with the driving dynamics in the sense of GS. By choosing appropriate coupling, the coupled system could be symmetric or asymmetric. Interestingly, it is found that the response system exhibits bistability in both cases. Three different types of bistable chaos have been identified. The crisis bifurcations which lead to the bistability are explored, and the relation between the bistable attractors is analyzed. The basin of attraction of the bistable attractors is extensively studied in both parameter space and initial condition space. The fractal basin boundary and the riddled basin are observed and they are characterized in terms of the uncertainty exponent. PMID- 15903549 TI - Time-independent approximations for periodically driven systems with friction. AB - The classical dynamics of a particle that is driven by a rapidly oscillating potential (with frequency omega) is studied. The motion is separated into a slow part and a fast part that oscillates around the slow part. The motion of the slow part is found to be described by a time-independent equation that is derived as an expansion in orders of omega(-1) (in this paper terms to the order omega(-3) are calculated explicitly). This time-independent equation is used to calculate the attracting fixed points and their basins of attraction. The results are found to be in excellent agreement with numerical solutions of the original time dependent problem. PMID- 15903550 TI - Lyapunov instabilities of Lennard-Jones fluids. AB - Recent work on many-particle systems reveals the existence of regular collective perturbations corresponding to the smallest positive Lyapunov exponents (LEs), called hydrodynamic Lyapunov modes. Until now, however, these modes have been found only for hard-core systems. Here we report results on Lyapunov spectra and Lyapunov vectors (LVs) for Lennard-Jones fluids. By considering the Fourier transform of the coordinate fluctuation density u((alpha)) (x,t) , it is found that the LVs with lambda approximately equal to 0 are highly dominated by a few components with low wave numbers. These numerical results provide strong evidence that hydrodynamic Lyapunov modes do exist in soft-potential systems, although the collective Lyapunov modes are more vague than in hard-core systems. In studying the density and temperature dependence of these modes, it is found that, when the value of the Lyapunov exponent lambda((alpha)) is plotted as function of the dominant wave number k(max) of the corresponding LV, all data from simulations with different densities and temperatures collapse onto a single curve. This shows that the dispersion relation lambda((alpha)) vs k(max) for hydrodynamical Lyapunov modes appears to be universal for the low-density cases studied here. Despite the wavelike character of the LVs, no steplike structure exists in the Lyapunov spectrum of the systems studied here, in contrast to the hard-core case. Further numerical simulations show that the finite-time LEs fluctuate strongly. We have also investigated localization features of LVs and propose a length scale to characterize the Hamiltonian spatiotemporal chaotic states. PMID- 15903551 TI - Phase synchronization of a pair of spiral waves. AB - The interaction of a pair of spiral waves with different independent rotation frequencies is studied. In a very large frequency mismatch searching region, we observe three different pattern formation phenomena: (a) phase-synchronization induced invasion under a relatively small frequency mismatch, i.e., the spiral wave with slower frequency (longer period) is swept away by a traveling wave, which is induced and phase synchronized by the faster spiral wave; (b) the coexistence of two spiral waves at sufficiently large parameter mismatch; and (c) an intermediate state, a non-phase-synchronous invasion, that is, similarly the slower spiral wave is swept by an approximate planar wave, whose frequency, however, is intermediate between those of the faster and slower waves. A point source model is studied to analyze all these phenomena in a unified way. PMID- 15903552 TI - Thermodynamic formalism for the Lorentz gas with open boundaries in d dimensions. AB - A Lorentz gas may be defined as a system of fixed dispersing scatterers, with a single light particle moving among these and making specular collisions on encounters with the scatterers. For a dilute Lorentz gas with open boundaries in d dimensions we relate the thermodynamic formalism to a random flight problem. Using this representation we analytically calculate the central quantity within this formalism, the topological pressure, as a function of system size and a temperaturelike parameter beta . The topological pressure is given as the sum of the topological pressure for the closed system and a diffusion term with a beta dependent diffusion coefficient. From the topological pressure we obtain the Kolmogorov-Sinai entropy on the repeller, the topological entropy, and the partial information dimension. PMID- 15903553 TI - Energy localization in two chaotically coupled systems. AB - We set up and analyze a random matrix model to study energy localization and its time behavior in two chaotically coupled systems. This investigation is prompted by a recent experimental and theoretical study of Weaver and Lobkis on coupled elastomechanical systems. Our random matrix model properly describes the main features of the findings by Weaver and Lobkis. Due to its general character, our model is also applicable to similar systems in other areas of physics--for example, to chaotically coupled quantum dots. PMID- 15903554 TI - Effective dynamics in Hamiltonian systems with mixed phase space. AB - An adequate characterization of the dynamics of Hamiltonian systems at physically relevant scales has been largely lacking. Here we investigate this fundamental problem and we show that the finite-scale Hamiltonian dynamics is governed by effective dynamical invariants, which are significantly different from the dynamical invariants that describe the asymptotic Hamiltonian dynamics. The effective invariants depend both on the scale of resolution and the region of the phase space under consideration, and they are naturally interpreted within a framework in which the nonhyperbolic dynamics of the Hamiltonian system is modeled as a chain of hyperbolic systems. PMID- 15903555 TI - Noise-induced instability: an approach based on higher-order moments. AB - Noise-induced transitions in the organization of systems far from equilibrium have been of vital interest. Although the effects of additive and multiplicative noise have been widely studied, it is only the multiplicative noise that can be dealt with within the scope of a linear analysis of first moments of the spatiotemporal perturbations, by the application of Novikov's theorem. For the case of additive noise, the corresponding straightforward linear analysis of the first moment throws no light on the effect of the noise on stability conditions. We propose here a simple approach based on higher-order moments to show how additive noise can give rise to noise-induced instability in spatially extended systems, at times leading to pattern formation. Our theoretical analysis is corroborated by numerical simulations on two simple one-component reaction diffusion systems in two dimensions. PMID- 15903556 TI - Synchrony of neural oscillators induced by random telegraphic currents. AB - When a neuron receives a randomly fluctuating input current, its reliability of spike generation improves compared with the case of a constant input current [Mainen and Sejnowski, Science 268, 1503 (1995)]. This phenomenon can be interpreted as phase synchronization between uncoupled nonlinear oscillators subject to a common external input. We analyze this phenomenon using dynamical models of neurons, assuming the input current to be a simple random telegraphic signal that jumps between two values, and the neuron to be always purely self oscillatory. The internal state of the neuron randomly jumps between two limit cycles corresponding to the input values, which can be described by random phase maps when the switching time of the input current is sufficiently long. Using such a random map description, we discuss the synchrony of neural oscillators subject to fluctuating inputs. Especially when the phase maps are monotonic, we can generally show that the Lyapunov exponent is negative, namely, phase synchronization is stable and reproducibility of spike timing improves. PMID- 15903557 TI - Lyapunov exponents from unstable periodic orbits. AB - We propose a method that allows us to analytically compute the largest Lyapunov exponent of a Hamiltonian chaotic system from the knowledge of a few unstable periodic orbits (UPOs). In the framework of a recently developed theory for Hamiltonian chaos, by computing the time averages of the metric tensor curvature and of its fluctuations along analytically known UPOs, we have re-derived the analytic value of the largest Lyapunov exponent for the Fermi-Pasta-Ulam-beta (FPU-beta) model. The agreement between our results and the Lyapunov exponents obtained by means of standard numerical simulations confirms the point of view which attributes to UPOs the special role of efficient probes of general dynamical properties, among them chaotic instability. PMID- 15903558 TI - Determinism test for very short time series. AB - A test for determinism suitable for time series shorter than 100 points is presented, and applied to numerical and observed data. The method exploits the linear d(d(0)) dependence in the expression d(t) approximately d(0)e(lambda t) which describes the growth of small separations between trajectories in chaotic systems. PMID- 15903559 TI - Delocalized and resonant quantum transport in nonlinear generalizations of the kicked rotor model. AB - We analyze the effects of a nonlinear cubic perturbation on the delta-kicked rotor. We consider two different models, in which the nonlinear term acts either in the position or in the momentum representation. We numerically investigate the modifications induced by the nonlinearity in the quantum transport in both localized and resonant regimes and a comparison between the results for the two models is presented. Analyzing the momentum distributions and the increase of the mean square momentum, we find that the quantum resonances asymptotically are very stable with respect to nonlinear perturbation of the rotor's phase evolution. For an intermittent time regime, the nonlinearity even enhances the resonant quantum transport, leading to superballistic motion. PMID- 15903560 TI - Mechanism for the partial synchronization in three coupled chaotic systems. AB - We investigate the dynamical mechanism for the partial synchronization in three coupled one-dimensional maps. A completely synchronized attractor on the diagonal becomes transversely unstable via a blowout bifurcation, and then a two-cluster state, exhibiting on-off intermittency, appears on an invariant plane. If the newly created two-cluster state is transversely stable, then partial synchronization occurs on the invariant plane; otherwise, complete desynchronization takes place. It is found that the transverse stability of the intermittent two-cluster state may be determined through the competition between its laminar and bursting components. When the laminar (bursting) component is dominant, partial synchronization (complete desynchronization) occurs through the blowout bifurcation. This mechanism for the occurrence of partial synchronization is also confirmed in three coupled multidimensional invertible systems, such as coupled He non maps and coupled pendula. PMID- 15903561 TI - Type specification of stability islands and chaotic stickiness. AB - A detailed characterization of stability islands in area-preserving maps is introduced on the basis of the resonance partition of phase space and it is used to define chaotic stickiness in these maps. It is shown that a general island can be characterized by a well-defined quasiregularity "type," specifying the sequence of resonances visited by the island. In particular, a "tangle" island lies entirely not just within the turnstile lobe of a resonance but also within the turnstile overlap of two resonances. Chaotic stickiness to a given island is then defined as the coincidence of the type of a chaotic orbit with that of the island in some time interval. This definition allows one to study stickiness systematically on all time scales, including short or nonasymptotic time regimes, as illustrated in the case of an accelerator-mode island of the standard map. A physically significant identification of the "sticky layer" and its "sublayers" in this case is made and discussed. PMID- 15903562 TI - Mesoscopic fluctuations of the Loschmidt echo. AB - We investigate the time-dependent variance of the fidelity with which an initial narrow wave packet is reconstructed after its dynamics is time reversed with a perturbed Hamiltonian. In the semiclassical regime of perturbation, we show that the variance first rises algebraically up to a critical time t(c) , after which it decays. To leading order in the effective Planck's constant Planck's(eff) , this decay is given by the sum of a classical term approximately same as exp [ 2lambdat] , a quantum term approximately same as 2Planck's(eff) exp [-Gamma t] , and a mixed term approximately 2 exp [- (Gamma+lambda) t] . Compared to the behavior of the average fidelity, this allows for the extraction of the classical Lyapunov exponent lambda in a larger parameter range. Our results are confirmed by numerical simulations. PMID- 15903563 TI - Spiral patterns, spiral breakup, and zigzag spirals in an optical device. AB - Above the first lasing threshold the degenerate optical parametric oscillator with saturable absorber displays successive Hopf and Turing instabilities. Various spiral patterns and defect turbulent patterns are numerically observed on the light intensity profiles. Close to the Hopf threshold, a normal form is derived which leads to a complex Ginzburg-Landau equation where a bi-Laplacian instead of a Laplacian drives the formation of spirals. At resonance the predictions of the normal form are compared with the numerical observations of the full equations. Above the Hopf threshold, the spirals destabilize, breaking into slowly evolving patterns with small spirals and filaments. Further above the threshold, when both the Turing and Hopf bifurcations interplay, a new spiral pattern emerges, with large notched arms. PMID- 15903564 TI - Symmetry breaking: a heuristic approach to chaotic scattering in many dimensions. AB - As the theory of chaotic scattering in high-dimensional systems is poorly developed, it is very difficult to determine initial conditions for which interesting scattering events, such as long delay times, occur. We propose to use symmetry breaking as a way to gain the insight necessary to determine low dimensional subspaces of initial conditions in which we can find such events easily. We study numerically the planar scattering off a disk moving on an elliptic Kepler orbit, as a simplified model of the elliptic restricted three body problem. When the motion of the disk is circular, the system has an integral of motion, the Jacobi integral, which is no longer conserved for nonvanishing eccentricity. In the latter case, the system has an effective five-dimensional phase space and is therefore not amenable for study with the usual methods. Using the symmetric problem as a starting point we define an appropriate two dimensional subspace of initial conditions by fixing some coordinates. This subspace proves to be useful to define scattering experiments where the rich and nontrivial dynamics of the problem is illustrated. We consider in particular trajectories which take very long before escaping or are trapped by consecutive collisions with the disk. PMID- 15903565 TI - Signal propagation and failure in one-dimensional FitzHugh-Nagumo equations with periodic stimuli. AB - We analyze the effect of additive periodic stimuli in one-dimensional FitzHugh Nagumo equations in an excitable regime. With a suitable stimulus interval, the suppression of the pulse propagation occurs in some parameter regime. This propagation failure comes from the formation of the "death spot" where successive pulses annihilate. In the parameter regime where the solitary pulse cannot propagate in space stably, however, periodic stimuli cause a propagation of envelope of a traveling pulse under a "resonance" condition, i.e., the pulse at the leading edge disappears successively, however, an envelope is formed and propagates with keeping its shape. PMID- 15903566 TI - Avoided level crossings, diabolic points, and branch points in the complex plane in an open double quantum dot. AB - We study the spectrum of an open double quantum dot as a function of different system parameters in order to receive information on the geometric phases of branch points in the complex plane (BPCP). We relate them to the geometrical phases of the diabolic points (DPs) of the corresponding closed system. The double dot consists of two single dots and a wire connecting them. The two dots and the wire are represented by only a single state each. The spectroscopic values follow from the eigenvalues and eigenfunctions of the Hamiltonian describing the double dot system. They are real when the system is closed, and complex when the system is opened by attaching leads to it. The discrete states as well as the narrow resonance states avoid crossing. The DPs are points within the avoided level crossing scenario of discrete states. At the BPCP, width bifurcation occurs. Here, different Riemann sheets evolve and the levels do not cross anymore. The BPCP are physically meaningful. The DPs are unfolded into two BPCP with different chirality when the system is opened. The geometric phase that arises by encircling the DP in the real plane, is different from the phase that appears by encircling the BPCP. This is found to be true even for a weakly opened system and the two BPCP into which the DP is unfolded. PMID- 15903567 TI - Critical behavior of a dynamic analog to the q=3 Potts model. AB - We construct a dynamical analog to the q=3 Potts model, using linear chaotic maps and a diffusive coupling. We find well-defined order-disorder phase transitions (PTs) in the system, and obtain the phase diagrams for both simultaneous and sequential updating of the model. For simultaneous updating we find continuous PTs whose critical exponents are consistent with those of the equilibrium Potts model. Under sequential updating, the phase diagram shows a tricritical point, and the PTs become first order for large coupling and chaoticity of the local maps. A preliminary estimation finds critical exponents in the region of continuous PTs that are not consistent with those of the equilibrium model. PMID- 15903568 TI - Self-collimation in liquid surface waves propagating over a bottom with periodically drilled holes. AB - Liquid surface waves propagating over a bottom with periodically drilled holes are studied experimentally and theoretically. Point source waves are generated in the center of the array of the periodically drilled holes in order to observe the evolvements of these waves. We successfully observe the self-collimation phenomenon in liquid surface waves. Band structures and constant-frequency surfaces are calculated, which can give a satisfactory interpretation of the experimental observations. PMID- 15903569 TI - Excitation of large-scale inertial waves in a rotating inhomogeneous turbulence. AB - A mechanism of excitation of the large-scale inertial waves in a rotating inhomogeneous turbulence due to an excitation of a large-scale instability is found. This instability is caused by a combined effect of the inhomogeneity of the turbulence and the uniform mean rotation. The source of the large-scale instability is the energy of the small-scale turbulence. We determined the range of parameters at which the large-scale instability occurs, the growth rate of the instability, and the frequency of the generated large-scale inertial waves. PMID- 15903570 TI - Inverse structure functions. AB - While the ordinary structure function in turbulence is concerned with the statistical moments of the velocity increment Deltau measured over a distance r , the inverse structure function is related to the distance r where the turbulent velocity exits the interval Deltau. We study inverse structure functions of wind tunnel turbulence which covers a range of Reynolds numbers Re(lambda) = 400-1100. We test a recently proposed relation between the scaling exponents of the ordinary structure functions and those of the inverse structure functions [S. Roux and M. H. Jensen, Phys. Rev. E 69, 16309 (2004)]. The relatively large range of Reynolds numbers in our experiment also enables us to address the scaling with Reynolds number that is expected to highlight the intermediate dissipative range. While we firmly establish the (relative) scaling of inverse structure functions, our experimental results fail both predictions. Therefore, the question of the significance of inverse structure functions remains open. PMID- 15903571 TI - Hydrodynamic dispersion in a hierarchical network with a power-law distribution of conductances. AB - Dispersion is studied on a two-dimensional hierarchical pore network with a power law distribution of conductances, i.e., P(g) approximately g(mu-1), with gepsilon(0,1), and mu is the disorderliness parameter (mu > 0). A procedure for computing tracer dispersion transport on hierarchical networks was developed. The results show that the effective diffusion coefficient of the network scales similarly as conduction on the same lattice. This means that the disorder length scales for conduction and diffusion processes are the same, and can be predicted from percolation theory. The dispersivity, xi identical with D(||)/U, was found to diverge rapidly as mu-->0. The result is in agreement with the model developed by Bouchaud and Georges (C.R. Acad. Sci. (Paris) 307 1431, 1988). A limiting value of mu approximately 0.45 was found, below which the convection-dispersion equation is no longer valid. PMID- 15903572 TI - Improved epsilon expansion for three-dimensional turbulence: two-loop renormalization near two dimensions. AB - An improved epsilon expansion in the d -dimensional (d > 2) stochastic theory of turbulence is constructed at two-loop order, which incorporates the effect of pole singularities at d--> 2 in coefficients of the epsilon expansion of universal quantities. For a proper account of the effect of these singularities, two different approaches to the renormalization of the powerlike correlation function of the random force are analyzed near two dimensions. By direct calculation, it is shown that the approach based on the mere renormalization of the nonlocal correlation function leads to contradictions at two-loop order. On the other hand, a two-loop calculation in the renormalization scheme with the addition to the force correlation function of a local term to be renormalized instead of the nonlocal one yields consistent results in accordance with the ultraviolet (UV) renormalization theory. The latter renormalization prescription is used for the two-loop renormalization-group analysis amended with partial resummation of the pole singularities near two dimensions, leading to a significant improvement of the agreement with experimental results for the Kolmogorov constant. PMID- 15903573 TI - Fastest growing linear Rayleigh-Taylor modes at solid/fluid and solid/solid interfaces. AB - Previous linear stability analyses of the Rayleigh-Taylor instability problem for elastic solids have been restricted to calculating the cutoff wavelength lambda(c) (zero growth rate) in the limit of Atwood number A of unity. Here, we rigorously derive the dispersion relations for solid/fluid and solid/solid interfaces and perform a systematic investigation to compute the most unstable modes (maximum growth rate) for all A. After rationalizing the dispersion relations into multivariable polynomials, we compute the physically meaningful wavelength lambda and growth rate sigma for all unstable disturbances as a function of the mechanical properties of the participating media (shear moduli, dynamic viscosity, and density contrast) and acceleration. It is shown that at these interfaces, the onset of instability can only arise via monotonically growing disturbances. For solid/fluid and solid/solid interfaces, the locus of the most unstable wavelength lambda(m) and growth rate sigma(m) pairs are calculated to cover the entire range of behavior in dimensionless space. We find that under certain conditions, at solid/fluid interfaces, two configurations with distinct A can have the same lambda(m) (a behavior that does not occur at solid/solid interfaces). In terms of estimating sigma(m), lambda(m), and lambda(c), the applicability of our results extends to layers of finite thickness h provided h > lambda/2. We suggest a plausible mechanism to explain the wavelength selection process in nominally smooth magnetically imploded liners observed in recent experiments. PMID- 15903574 TI - Drag reduction in the turbulent Kolmogorov flow. AB - We investigate the phenomenon of drag reduction in a viscoelastic fluid model of dilute polymer solutions. By means of direct numerical simulations of the three dimensional turbulent Kolmogorov flow we show that drag reduction takes place above a critical Reynolds number Re(c). An explicit expression for the dependence of Re(c) on polymer elasticity and diffusivity is derived. The values of the drag coefficient obtained for different fluid parameters collapse onto a universal curve when plotted as a function of the rescaled Reynolds number Re/ Re(c). The analysis of the momentum budget allows us to gain some insight on the physics of drag reduction, and suggests the existence of a Re-independent value of the drag cofficient--lower than the Newtonian one--for large Reynolds numbers. PMID- 15903575 TI - Model of single bubble sonoluminescence. AB - The temperature within and the sonoluminescence characteristics of a stable inert gas single bubble grown in water under some given conditions are computed by using a model that is as sufficiently complete as we can manage, except that possible chemical reactions within the bubble are neglected. We work with several different versions of the equation describing the motion of the bubble wall, which are usually considered to give merely slight differences; or vary a parameter in the formula calculating the net increment of the water condensed at the bubble wall. It is found that the final outcomes of the temperature and the sonoluminescence can be significantly different in some cases. This illustration points to the importance of differentiating among the various seemingly similar equations and of adopting the correct value of the parameter used in the computation model of a single bubble. PMID- 15903576 TI - Complex flow around a bubble rising in a non-Newtonian fluid. AB - Our experimental investigation by both particle image velocimetry and birefringence modulation method shows very complex flow features around a bubble rising in a non-Newtonian fluid. We model this two-phase flow by coupling the free-energy-based lattice Boltzmann scheme and the fluid rheology in the framework of the sixth-order Maxwell model with shear thinning effects. A Newtonian low viscosity drop is used to simulate the rising bubble. Numerical results including noticeably negative wake behind the bubble, stress field, as well as the bubble's teardrop shape are obtained, and compare satisfactorily with our experiments. PMID- 15903577 TI - Long-wavelength equation for vertically falling films. AB - An equation is derived for describing wave evolution on the surface of a vertically falling viscous film. The traditional long-wavelength scaling is replaced by a new scaling to reduce the (We) must be used instead of the Reynolds number (Re) to distinguish between viscous and inertia dominated regimes for vertically falling films. This equation includes viscous dissipation and pressure correction terms that are missing in the existing single evolution equations at the same order. Comparison of the neutral stability curves and growth rates predicted by different models to that of the Orr-Sommerfeld (OS) equation shows that our equation matches with the OS results better than the existing single evolution equations. However, our equation is not free from finite time blowup. Selective regularization leads to a two mode model in flow rate and film thickness. The regularized equation is free from finite time blowup and predicts two families of solitary waves. Numerical simulations of the derived equation and its regularized version in the traveling wave coordinate show the transition of wave structure from regular (periodic) to chaotic profiles. Model predictions on maximum wave amplitude on the low celerity branch show good agreement with experimental data. PMID- 15903578 TI - Thermodynamics of magnetohydrodynamic flows with axial symmetry. AB - We present strategies based upon optimization principles in the case of the axisymmetric equations of magnetohydrodynamics (MHD). We derive the equilibrium state by using a minimum energy principle under the constraints of the MHD axisymmetric equations. We also propose a numerical algorithm based on a maximum energy dissipation principle to compute in a consistent way the nonlinearly dynamically stable equilibrium states. Then, we develop the statistical mechanics of such flows and recover the same equilibrium states giving a justification of the minimum energy principle. We find that fluctuations obey a Gaussian shape and we make the link between the conservation of the Casimirs on the coarse-grained scale and the process of energy dissipation. We contrast these results with those of two-dimensional hydrodynamical turbulence where the equilibrium state maximizes a H function at fixed energy and circulation and where the fluctuations are nonuniversal. PMID- 15903579 TI - Onset of convection in colloids stratified by gravity. AB - Settling of the equilibrium (barometric) distribution of colloidal grains in a liquid requires a very long time in comparison with that necessary for temperature equilibrium establishment. This mismatch enables different scenarios for the onset of thermal convection. If the stratified state has had no time to be settled, the colloid behaves as a pure fluid where only stationary convection arises. On the contrary, if the barometric concentration profile has been reached before the temperature gradient is imposed, only oscillatory convection occurs. Thereafter, the oscillations last as long as it takes the convection to wash away the initial concentration profile. Then the fluid becomes homogeneous and stationary convection is eventually established. The influence of the Soret effect upon the onset of convection is also taken into account and discussed. PMID- 15903580 TI - Evaporative deposition patterns: spatial dimensions of the deposit. AB - A model accounting for the finite spatial dimensions of the deposit patterns in evaporating sessile drops of a colloidal solution on a plane substrate is proposed. The model is based on the assumption that the solute particles occupy finite volume and hence these dimensions are of steric origin. Within this model, the geometrical characteristics of the deposition patterns are found as functions of the initial concentration of the solute, the initial geometry of the drop, and the time elapsed from the beginning of the drying process. The model is solved analytically for small initial concentrations of the solute and numerically for arbitrary initial concentrations of the solute. The agreement between our theoretical results and the experimental data is demonstrated, and it is shown that the observed dependence of the deposit dimensions on the experimental parameters can indeed be attributed to the finite dimensions of the solute particles. These results are universal and do not depend on any free or fitting parameters; they are important for understanding evaporative deposition and may be useful for creating controlled deposition patterns. PMID- 15903581 TI - Single-mode dynamics of the Rayleigh-Taylor instability at any density ratio. AB - The behavior of a periodic array of Rayleigh-Taylor bubbles (and spikes) of wavelength lambda is investigated at different density ratios using three dimensional numerical simulations. The scaled bubble and spike velocities (v(b,s)/sqrt[Aglambda/2]), are found to vary with the Atwood number A, and are compared with recent potential flow theories. Simulations at different grid resolutions reveal that the convergence rates of bubble velocities improve with increasing A, while the converse holds true for spike velocities. The asymptotic radius of curvature at the bubble tip is found to be independent of A, consistent with potential flow theory. These results are useful in validating potential flow theory models of the nonlinear stage of the Rayleigh-Taylor instability. PMID- 15903582 TI - Dielectric matrix and plasmon dispersion in strongly coupled electronic bilayer liquids. AB - We develop a dielectric matrix and analyze plasmon dispersion in strongly coupled charged-particle bilayers in the T = 0 quantum domain. The formulation is based on the classical quasilocalized charge approximation (QLCA) and extends the QLCA formalism into the quantum domain. Its development, which parallels that of the two-dimensional companion paper [Phys. Rev. E 70, 026406 (2004)] by three of the authors, generalizes the single-layer scalar formalism therein to a bilayer matrix formalism. Using pair correlation function data generated from diffusion Monte Carlo simulations, we calculate the dispersion of the in-phase and out-of phase plasmon modes over a wide range of high- r(s) values and layer separations. The out-of-phase spectrum exhibits an exchange-correlation induced long wavelength energy gap in contrast to earlier predictions of acoustic dispersion softened by exchange and correlations. The energy gap is similar to what has been previously predicted for classical charged-particle bilayers and subsequently confirmed by recent molecular dynamics computer simulations. PMID- 15903583 TI - Comparison of experimental and simulated extreme ultraviolet spectra of xenon and tin discharges. AB - Xenon and tin both are working elements applied in discharge plasmas that are being developed for application in extreme ultraviolet (EUV) lithography. Their spectra in the 10-21-nm-wavelength range have been analyzed. A fully analytical collisional-radiative model, including departure from equilibrium due to a net ionization rate, was used to simulate the EUV spectra. Detailed Hartree-Fock calculations, using the COWAN package, were applied for determination of the energy levels and optical transition probabilities of the 8+ to 12+ ions of both elements. For the calculation of the radiation, the opacity of the plasma was taken into account. Time-resolved measurements of the spectra from ionizing phases of two different discharge plasmas were corrected for the wavelength dependent sensitivity of the spectrometer, and compared to the results of the simulations. Fairly good agreement between the experiments and the model calculations has been found. PMID- 15903584 TI - Enhancement of energetic electrons and protons by cone guiding of laser light. AB - Energetic electrons and protons are observed when a target consisting of a reentrant cone with a disk at the tip is irradiated by a petawatt (PW) laser at an intensity of approximately 10(19) W cm(-2). The angular distribution of the electrons and protons, dependent on the open angle of the reentrant cone, is found to differ from that in the case when a target with planar geometry is used. Two jet beams are observed, in directions parallel to the cone axis and normal to the cone-shaped wall. The number and cutoff energies of the generated protons are also related to the open angle of the cone. The efficiency of the generation of energetic electrons from the cone target is 2-3 times higher than that from a simple plane target. These results indicate a guiding of the PW laser beam in the cone geometry. PMID- 15903585 TI - Attosecond electromagnetic pulse generation due to the interaction of a relativistic soliton with a breaking-wake plasma wave. AB - During the interaction of a low-frequency relativistic soliton with the electron density modulations of a wake plasma wave, part of the electromagnetic energy of the soliton is reflected in the form of an extremely short and ultraintense electromagnetic pulse. We calculate the spectra of the reflected and of the transmitted electromagnetic pulses analytically. The reflected wave has the form of a single cycle attosecond pulse. PMID- 15903586 TI - Measured and calculated SF-6 collision and swarm ion transport data in SF6 -Ar and SF6 -Xe mixtures. AB - The measurement of the mobility of SF-6 in the mixtures SF6 -Ar and SF6 -Xe is reported over the density-reduced electric field strength E/N 1-180 Td (1 Townsend = 10(-17) V cm(2)), from a time-resolved pulsed Townsend technique. Simultaneously, the mobility of SF-6 in the same binary mixtures has been calculated from a set of collision cross sections for SF-6 -Ar, SF-6 -Xe, and SF 6 - SF6 using a Monte Carlo simulation procedure for ion transport. The good agreement between measured and calculated mobilities in these gas mixtures has led us to conclude that the validation of our cross section sets is confirmed. The elastic collision cross section, a predominant process for ion energies lower than about 10 eV, was determined from a semiclassical JWKB approximation using a rigid core potential model for the ion-neutral systems under consideration. This elastic cross section was then added to several other inelastic collision cross sections found in the literature for ion conversion, electron detachment of SF-6 and charge transfer. Moreover, the calculations of the mobility and the ratios of the transverse and longitudinal diffusion coefficients to the mobility were extended into a much wider E/N range from 1 to 4000 Td. Additionally, we have also calculated the energy distribution functions and the reaction coefficients for ion conversion and electron detachment. Finally, we have shown that the range of validity for the calculation of the mobility in gas mixtures from Blanc's law is only valid for the low E/N region, where the interaction is dominated by elastic collisions and the ion distribution function remains essentially Maxwellian. PMID- 15903587 TI - Longitudinal cooling of non-neutral plasma by energy exchange. AB - The optimal values of Q and Deltaomega (Deltaomega identical withomega-Omega) for cooling a pure electron plasma with a microwave bath have been calculated. An electron plasma, which has no internal degree of freedom, cannot be cooled below the temperature of a heat bath. However, longitudinal cooling can be achieved by energy transfer from the poorly cooled longitudinal degree of freedom to the well cooled (by synchrotron radiation) transverse degree of freedom. To do this, a microwave bath is introduced to the electron plasma. A microwave tuned to a frequency below the gyrofrequency forces electrons moving towards the microwave to absorb a microwave photon. The electrons move up one in Landau state and then lose their longitudinal momenta. In this process, the longitudinal temperature of the electron plasma decreases. On the basis that the perpendicular temperature is below the Landau temperature of the plasma, we set up two level transition equations and then derive a Fokker-Planck equation from them. With the aid of a finite element method (FEM) code for the equation, the cooling times for several values of the magnetic field, the microwave cavity (Q), and the relative detuning frequency from the gyrofrequency (Deltaomega) are calculated. Thus optimal values of the microwave cavity and the detuning frequency for longitudinal cooling of a strongly magnetized electron plasma with a microwave bath have been found. By applying these optimal values with an appropriate microwave intensity, the best cooling can be obtained. For an electron plasma magnetized to 10 T, the cooling time to the solid state is approximately two hours. PMID- 15903588 TI - Asymmetry of Hbeta Stark profiles in T-tube hydrogen plasma. AB - The whole Balmer H(beta) line profiles are studied in detail experimentally in the T-tube discharge for the wide range of plasma parameters. Besides the common one, two additional parameters are introduced to characterize the asymmetry behavior of the experimental Stark profiles with the reference point chosen in the center of the line. The experimental data are analyzed and benchmarked versus the simple theoretical model based on the effects of microfield nonuniformity and electron impact shifts. PMID- 15903589 TI - Semiclassical model for the ionic self-diffusion coefficient in white dwarfs. AB - Under the extreme conditions of massive white dwarfs, which have ionic densities that exceed 10(29) cm(-3) , the ions can be both very strongly coupled and partially degenerate. We present a simple model for self-diffusion in such white dwarfs that utilizes the known one-component plasma diffusion coefficient and scalings derived from the short-time expansions of the velocity autocorrelation function and the memory function. Since the ions are weakly degenerate, we utilize a simple semiclassical correction to the classical dynamics. We find enhanced diffusion, relative to the purely classical calculation, which is more significant at smaller values of the Coulomb coupling parameter. PMID- 15903590 TI - Numerical investigation of the discharge characteristics of the pulsed discharge nozzle. AB - The characteristics of the plasma generated by a pulsed discharge slit nozzle (PDN) are investigated. The PDN source is designed to produce and cool molecular ions creating an astrophysically relevant environment in the laboratory. A discharge model is applied to this system to provide a qualitative as well as a quantitative picture of the plasma. We find that the plasma's properties and behavior are characteristic of those of a glow discharge. We model the electron density and energy, as well as the argon ion and metastable atom number density. The results reveal a high abundance of metastable argon atoms in the expansion region, which is more than one order of magnitude higher than the abundance of electrons and ions. These findings confirm experimental observations, which concluded that large molecular ions are dominantly formed through Penning ionization of the neutral molecular precursors seeded in the supersonic expansion of argon gas. The simulations presented here will help optimize the yield of formation of molecular ions and radicals in the PDN source; they will also provide key physical insight into the characteristics of interstellar molecules and ions analogs in laboratory experiments. PMID- 15903591 TI - Relativistic electromagnetic ion cyclotron instabilities. AB - The relativistic instabilities of electromagnetic ion cyclotron waves driven by MeV ions are analytically and numerically studied. As caused by wave magnetic field and in sharp contrast to the electrostatic case, interesting characteristics such as Alfve nic behavior and instability transition are discovered and illuminated in detail. The instabilities are reactive and are raised from the coupling of slow ions' first-order resonance and fast ions' second-order resonance, that is an essential extra mechanism due to relativistic effect. Because of the wave magnetic field, the nonresonant plasma dielectric is usually negative and large, that affects the instability conditions and scaling laws. A negative harmonic cyclotron frequency mismatch between the fast and slow ions is required for driving a cubic (and a coupled quadratic) instability; the cubic (square) root scaling of the peak growth rate makes the relativistic effect more important than classical mechanism, especially for low fast ion density and Lorentz factor being close to unity. For the cubic instability, there is a threshold (ceiling) on the slow ion temperature and density (the external magnetic field and the fast ion energy); the Alfve n velocity is required to be low. This Alfve nic behavior is interesting in physics and important for its applications. The case of fast protons in thermal deuterons is numerically studied and compared with the analytical results. When the slow ion temperature or density (the external magnetic field or the fast ion energy) is increased (reduced) to about twice (half) the threshold (ceiling), the same growth rate peak transits from the cubic instability to the coupled quadratic instability and a different cubic instability branch appears. The instability transition is an interesting new phenomenon for instability. PMID- 15903592 TI - Modeling of clusters in a strong 248-nm laser field by a three-dimensional relativistic molecular dynamic model. AB - A relativistic time-dependent three-dimensional particle simulation model has been developed to study the interaction of intense ultrashort KrF (248 nm) laser pulses with small Xe clusters. The trajectories of the electrons and ions are treated classically according to the relativistic equation of motion. The model has been applied to a different regime of ultrahigh intensities extending to 10(21) W/ cm(2). In particular, the behavior of the interaction with the clusters from intensities of approximately 10(15) W/cm(2) to intensities sufficient for a transition to the so-called "collective oscillation model" has been explored. At peak intensities below 10(20) W/cm(2), all electrons are removed from the cluster and form a plasma. It is found that the "collective oscillation model" commences at intensities in excess of 10(20) W/cm(2), the range that can be reached in stable relativistic channels. At these high intensities, the magnetic field has a profound effect on the shape and trajectory of the electron cloud. Specifically, the electrons are accelerated to relativistic velocities with energies exceeding 1 MeV in the direction of laser propagation and the magnetic field distorts the shape of the electron cloud to give the form of a pancake. PMID- 15903593 TI - Coulomb explosion effect and the maximum energy of protons accelerated by high power lasers. AB - The acceleration of light ions (protons) through the interaction of a high-power laser pulse with a double-layer target is theoretically studied by means of two dimensional particle-in-cell simulations and a one-dimensional analytical model. It is shown that the maximum energy acquired by the accelerated light ions (protons) depends on the physical characteristics of a heavy-ion layer (electron ion mass ratio and effective charge state of the ions). In our theoretical model, the hydrodynamic equations for both electron and heavy-ion species are solved and the test-particle approximation for the light ions (protons) is applied. The heavy-ion motion is found to modify the longitudinal electric field distribution, thus changing the acceleration conditions for the protons. PMID- 15903594 TI - Shock wave formation in a dc glow discharge dusty plasma. AB - A jump of dust density propagating through the dusty plasma structure has been observed. To excite the disturbance an impulse of axial magnetic field to the dusty plasma in a dc glow discharge striation has been applied. This impulse resulted in the dynamical stretching of the dusty plasma structure. During the reconstruction of the structure a ramp-shaped perturbation of dust density appeared. The perturbation was steepening and formed into a dust-acoustic shock. The anomalously high shock compression is observed. PMID- 15903595 TI - Laser heating of particles in dusty plasmas. AB - Experiments on the heating and melting of two-dimensional finite dust crystals are performed using random laser excitation of the dust particles by a rapidly moving laser beam. The achievable dust temperatures scale with the square of the laser power. The heating process is described for different dust clusters under various plasma and cluster conditions. A single-particle model is developed to explain the observed behavior of the cluster under the random laser excitation. Good quantitative agreement is found when the radiation pressure is made responsible for the particle excitation by the laser. The dynamical properties of the system during heating are analyzed and the dominant modes are identified. From this, it is demonstrated that the heating process is of a nearly equilibrium nature in contrast to previous melting experiments. Finally, the melting of the dust cluster by laser heating is studied. From these experiments, a precise determination of the critical coupling parameter for the solid-fluid transition was possible. It is measured as Gamma = 270-480 for an N = 18 cluster. PMID- 15903596 TI - Coherent beam-beam tune shift of unsymmetrical beam-beam interactions with large beam-beam parameter. AB - Coherent beam-beam tune shift of unsymmetrical beam-beam interactions was studied experimentally and numerically in HERA, where the lepton beam has such a large beam-beam parameter (up to xi(y) = 0.272 ) that the single-particle motion is locally unstable at the origin (closed orbit). Unlike the symmetrical case of beam-beam interactions, the ratio of the coherent beam-beam tune shift and the beam-beam parameter in this unsymmetrical case of beam-beam interactions was found to decrease monotonically with an increase of the beam-beam parameter. The results of self-consistent beam-beam simulation, the linearized Vlasov equation, and the rigid-beam model were compared with the experimental measurement. It was found that the coherent beam-beam tune shifts measured in the experiment and calculated in the simulation agree remarkably well but they are much smaller than those calculated by the linearized Vlasov equation with the single-mode approximation or the rigid-beam model. The study indicated that the single-mode approximation in the linearization of the Vlasov equation is not valid in the case of unsymmetrical beam-beam interactions. The rigid-beam model is valid only with a small beam-beam parameter in the case of unsymmetrical beam-beam interactions. PMID- 15903597 TI - Formulas for tune shift and beta beat due to perturbations in circular accelerators. AB - Rigorous formulas for nonlinear tune shift and beta -function distortion due to perturbations in the focusing forces are presented, which complement the well known tune-shift formula for quadrupole errors. Using these formulas, the calculation of nonlinear chromaticity given by Takao [Phys. Rev. E 70, 016501 (2004)] can be greatly simplified and extended to higher order. In addition, an expression for the nonlinear chromatic beta -function distortion is given. PMID- 15903598 TI - Space-frequency model of ultrawide-band interactions in free-electron lasers. AB - The principle of operation of intense radiation devices such as microwave tubes, free-electron lasers, and masers, is based on a distributed interaction between an electron beam and electromagnetic radiation. Some of the effects emerging during the interaction involve a continuum of frequencies in their broadband spectrum. We developed a three-dimensional, space-frequency theory for the analysis and simulation of radiation excitation and propagation in electron devices and free-electron lasers operating in an ultrawide range of frequencies. The total electromagnetic field (radiation and space-charge waves) is presented in the frequency domain as an expansion in terms of transverse eigenmodes of the (cold) cavity, in which the field is excited and propagates. The mutual interaction between the electron beam and the electromagnetic field is fully described by coupled equations, expressing the evolution of mode amplitudes and electron beam dynamics. The approach is applied in a numerical particle code WB3D, simulating wideband interactions in free-electron lasers operating in the linear and nonlinear regimes. PMID- 15903599 TI - Stimulated Raman scattering with strong damping: a simple theory of the spike phenomenon. AB - The classical stimulated Raman scattering system describing the resonant interaction between two electromagnetic waves and a fast relaxing medium wave is studied by introducing a systematic perturbation approach in powers of the relaxation time. We separate amplitude and phase effects for these complex fields. The analysis of the former shows the existence of a stagnation distance after which monotonic energy transfer begins from one electromagnetic wave to the other, and this quantity is calculated. Concerning phase effects we give the conditions for the formation of a Raman spike from an initial fast and large phase jump in one of the waves. The spike evolution and width estimated from the reduced model agree with the results from numerical simulations of the original system. PMID- 15903600 TI - Kink shape solutions of the Maxwell-Lorentz system. AB - In the limit of high amplitude oscillating electromagnetic fields, a sequence of kink antikink shaped optical waves has been found in the Maxwell's equations coupled to a single Lorentz oscillator and with Kerr nonlinearity. The individual kinks and antikinks result from a traveling wave assumption and their stability has been assessed by numerical simulations. For typical physical parameter values the kink width is of the order of tens of femtoseconds. PMID- 15903601 TI - Acoustic behavior of ordered droplets in a liquid: a phase space approach. AB - The transmission of an acoustical signal through a spatial arrangement consisting of a bidimensional crystal of droplets (liquid spheres) immersed into another liquid is analyzed. As a first approximation, the paraxial case is solved by considering a set of acoustical lenses which allow us to model the effect of each droplet on the signal. An expression for the Wigner distribution function that lets us evaluate the corresponding image, diffraction pattern, and even the output signal of any given paraxial input signal to that crystalline substrate is obtained, with particular emphasis on the case of an incoming plane wave. To solve the nonparaxial situation, a generalization of the concept of focal distance interpreting every sphere as a superposition of concentric rings of different radius, which permits us to find a general expression for the Wigner distribution function is proposed. PMID- 15903602 TI - Interactions among periodic waves and solitary waves of the (N + 1)-dimensional sine-Gordon field. AB - Exact solutions of the (n + 1)-dimensional sine-Gordon field equation are studied with help of those of the cubic nonlinear Klein-Gordon fields. The mapping relations among the sine-Gordon field equation and the cubic nonlinear Klein Gordon fields are pure algebraic. By solving the cubic nonlinear Klein-Gordon equations, many new types of exact explicit solutions such as the periodic periodic interaction waves, periodic-kink interaction waves, periodic perturbed "snake" shape solitary waves, etc., are displayed both analytically and graphically. PMID- 15903603 TI - Slanted-pore photonic band-gap materials. AB - We present a detailed analysis of three-dimensional photonic band-gap materials consisting of lattices of slanted pores with n = 2 , 3, and 4 pores per unit cell. These slanted pores emanate from one or two masks consisting of a two dimensional square or triangular lattice of holes placed on the top surface of the crystal. We also consider the case in which the top surface is "polished" at an angle after the first set of slanted pores is generated from the first mask. A second set of slanted pores is then generated from a second mask placed on the new angled surface. Using these architectures, we demonstrate photonic band gaps of up to 25% of the gap center frequency when the crystal is made from materials with a dielectric constant ratio of 11.9:1 (silicon with air pores). The proposed structures are amenable to microfabrication using established techniques such as x-ray lithography or two-photon direct writing in a polymer template, followed by inversion and replication of the template with polycrystalline silicon. They can also be microfabricated by direct plasma etching in single crystal semiconductors. An alternative fabrication protocol consists of photoelectrochemical etching of deep pores in a single crystal semiconductor followed by focused ion beam or reactive ion etching of the second lattice of directed pores. PMID- 15903604 TI - Coherence of light scattered from a randomly rough surface. AB - We study the coherence of p-polarized light scattered from a one-dimensional weakly rough random metal surface in contact with vacuum. The mutual coherence function of the single nonzero component of the scattered magnetic field is calculated in planes parallel to, and at increasing distances from, the mean scattering surface in the vacuum region. It is found to be the sum of a contribution that is independent of the distance from the mean surface and a contribution that is a function of this distance and decays to zero over a distance of the order of the wavelength of the incident light. It is also shown that the spatial coherence of the electromagnetic field in the far field in a plane at a fixed distance from the mean surface, as a function of the relative distance along it, mimics the surface height autocorrelation function at short relative distances and oscillates with two periods, T(1) = lambda and T(2) = lambda/sin theta(0), where theta(0) is the angle of incidence. The former is due to the excitation of lateral waves, while the latter is due to the coherent interference of the multiple scattering processes that lead to the enhanced backscattering effect. In the near field the spatial coherence of the electromagnetic field measured at a fixed distance from the mean surface displays oscillations that are due to the excitation of surface plasmon polaritons. The period of these oscillations equals the wavelength of the surface plasmon polaritons, while the exponential decay of their amplitude is determined by the energy mean free path of the surface plasmon polaritons. PMID- 15903605 TI - Full band gap for surface acoustic waves in a piezoelectric phononic crystal. AB - A plane-wave-expansion method suited to the analysis of surface-acoustic-wave propagation in two-dimensional piezoelectric phononic crystals is described. The surface modes of a square-lattice Y-cut lithium niobate phononic crystal with circular void inclusions with a filling fraction of 63% are identified. It is found that a large full band gap with a fractional bandwidth of 34% exists for surface acoustic waves of any polarization and incidence, coincidentally with the full band gap for bulk waves propagating in the plane of the surface. The excitation of surface acoustic waves by interdigital transducers is discussed. PMID- 15903606 TI - Quasistable two-dimensional solitons with hidden and explicit vorticity in a medium with competing nonlinearities. AB - We consider basic types of two-dimensional (2D) vortex solitons in a three-wave model combining quadratic chi((2)) and self-defocusing cubic chi((3))(-) nonlinearities. The system involves two fundamental-frequency (FF) waves with orthogonal polarizations and a single second-harmonic (SH) one. The model makes it possible to introduce a 2D soliton, with hidden vorticity (HV). Its vorticities in the two FF components are S(1,2) = +/-1 , whereas the SH carries no vorticity, S(3) = 0 . We also consider an ordinary compound vortex, with 2S(1) = 2S(2) = S(3) = 2 . Without the chi((3))(-) terms, the HV soliton and the ordinary vortex are moderately unstable. Within the propagation distance z approximately 15 diffraction lengths, Z(diffr), the former one turns itself into a usual zero-vorticity (ZV) soliton, while the latter splits into three ZV solitons (the splinters form a necklace pattern, with its own intrinsic dynamics). To gain analytical insight into the azimuthal instability of the HV solitons, we also consider its one-dimensional counterpart, viz., the modulational instability (MI) of a one-dimensional CW (continuous-wave) state with "hidden momentum," i.e., opposite wave numbers in its two components, concluding that such wave numbers may partly suppress the MI. As concerns analytical results, we also find exact solutions for spreading localized vortices in the 2D linear model; in terms of quantum mechanics, these are coherent states with angular momentum (we need these solutions to accurately define the diffraction length of the true solitons). The addition of the chi((3))(-) interaction strongly stabilizes both the HV solitons and the ordinary vortices, helping them to persist over z up to 50 Z(diffr). In terms of the possible experiment, they are completely stable objects. After very long propagation, the HV soliton splits into two ZV solitons, while the vortex with S(3) = 2S(1,2) = 2 splits into a set of three or four ZV solitons. PMID- 15903607 TI - Gradient index metamaterials. AB - Metamaterials--artificially structured materials with tailored electromagnetic response--can be designed to have properties difficult or impossible to achieve with traditional materials fabrication methods. Here we present a structured metamaterial, based on conducting split ring resonators (SRRs), which has an effective index of refraction with a constant spatial gradient. We experimentally confirm the gradient by measuring the deflection of a microwave beam by a planar slab of the composite metamaterial over a range of microwave frequencies. The gradient index metamaterial may prove an advantageous alternative approach to the development of gradient index lenses and similar optics, especially at higher frequencies. In particular, the gradient index metamaterial we propose may be suited for terahertz applications, where the magnetic resonant response of SRRs has recently been demonstrated. PMID- 15903608 TI - Mechanisms supporting long propagation regimes of photorefractive solitons. AB - We extend investigation of one-dimensional solitons in biased photorefractive crystals to long propagation regimes, where self-trapping over a large number of linear diffraction lengths combines with the progressive growth of generally distortive spatially nonlocal components. Results indicate that saturation halts the radiative misshaping of the soliton, which follows that specific bending trajectory along which its evolution is governed by the same local screening nonlinearity that intervenes in short propagation conditions, where spatial nonlocality has a negligible effect. This finding not only allows the prediction of the curvature and of the relative role of charge displacement and diffusion, but implies a set of interesting observable effects, such as boomerangs, counterpropagating and cavity geometries, quasirectilinear and anomalous collisions, along with specific consequences on soliton arrays and on coupling to bulk gratings. PMID- 15903609 TI - Peeling off an elastica from a smooth attractive substrate. AB - Using continuum mechanics, we study theoretically the unbinding of an inextensible rod with free ends attracted by a smooth substrate and submitted to a vertical force. We use the elastica model in a medium-range van der Waals potential. We numerically solve a nonlinear boundary value problem and obtain the force-stretching relation at zero temperature. We obtain the critical force for which the rod unbinds from the substrate as a function of three dimensionless parameters, and we find two different regimes of adhesion. We study analytically the contact potential case as the van der Waals radius goes to zero. PMID- 15903610 TI - Frozen light in periodic stacks of anisotropic layers. AB - We consider a plane electromagnetic wave incident on a periodic stack of dielectric layers. One of the alternating layers has an anisotropic refractive index with an oblique orientation of the principal axis relative to the normal to the layers. It was shown recently that an obliquely incident light, upon entering such a periodic stack, can be converted into an abnormal axially frozen mode with drastically enhanced amplitude and zero normal component of the group velocity. The stack reflectivity at this point can be very low, implying nearly total conversion of the incident light into the frozen mode with huge energy density, compared to that of the incident light. Supposedly, the frozen mode regime requires strong birefringence in the anisotropic layers--by an order of magnitude stronger than that available in common anisotropic dielectric materials. In this paper we show how to overcome the above problem by exploiting higher frequency bands of the photonic spectrum. We prove that a robust frozen mode regime at optical wavelengths can be realized in stacks composed of common anisotropic materials, such as YVO(4), LiNbO(3), CaCO(3), and the like. PMID- 15903611 TI - Mode-locking of mobile discrete breathers. AB - We study numerically synchronization phenomena of mobile discrete breathers in dissipative nonlinear lattices periodically forced. When varying the driving intensity, the breather velocity generically locks at rational multiples of the driving frequency. In most cases, the locking plateau coincides with the linear stability domain of the resonant mobile breather and desynchronization occurs by the regular appearance of type-I intermittencies. However, some plateaus also show chaotic mobile breathers with locked velocity in the locking region. The addition of a small subharmonic driving tames the locked chaotic solution and enhances the stability of resonant mobile breathers. PMID- 15903612 TI - Coupled nonlinear Schrodinger field equations for electromagnetic wave propagation in nonlinear left-handed materials. AB - For isotropic and homogeneous nonlinear left-handed materials, for which the effective medium approximation is valid, Maxwell's equations for electric and magnetic fields lead naturally, within the slowly varying envelope approximation, to a system of coupled nonlinear Schrodinger equations. This system is equivalent to the well-known Manakov model that under certain conditions, is completely integrable, and admits bright and dark soliton solutions. It is demonstrated that left- and right-handed (normal) nonlinear media may have compound dark and bright soliton solutions, respectively [corrected] These results are also supported by numerical calculations. PMID- 15903613 TI - Parametric light bullets supported by quasi-phase-matched quadratically nonlinear crystals. AB - We present a comprehensive analysis of the dynamics of three-dimensional spatiotemporal nonspinning and spinning solitons in quasi-phased-matched (QPM) gratings. By employing an averaging approach based on perturbation theory, we show that the soliton's stability is strongly affected by the QPM-induced third order nonlinearity (which is always of a mixed type, with opposite signs in front of the corresponding self-phase and cross-phase modulation terms). We study the dependence of the stability of the spatiotemporal soliton (STS) on its energy, spin, the wave-vector mismatch between the fundamental and second harmonics, and the relative strength of the intrinsic quadratic and QPM-induced cubic nonlinearities. In particular, all the spinning solitons are unstable against fragmentation, while zero-spin STS's have their stability regions on the system's parameter space. PMID- 15903614 TI - Combined solitary wave solutions for the inhomogeneous higher-order nonlinear Schrodinger equation. AB - We consider the inhomogeneous higher-order nonlinear Schrodinger equation and explicitly present exact combined solitary wave solutions that can describe the simultaneous propagation of bright and dark solitary waves in a combined form in inhomogeneous fiber media or in optical communication links with distributed parameters. Furthermore, we analyze the features of the solutions, and numerically discuss the stabilities of these solitary waves under slight violations of the parameter conditions and finite initial perturbations. The results show that there exist combined solitary wave solutions in an inhomogeneous fiber system, and the combined solitary wave solutions are stable under slight violations of the parameter conditions and finite initial perturbations. Finally, the interaction between two neighboring combined solitary waves is numerically discussed. PMID- 15903615 TI - Electromagnetic parameter retrieval from inhomogeneous metamaterials. AB - We discuss the validity of standard retrieval methods that assign bulk electromagnetic properties, such as the electric permittivity epsilon and the magnetic permeability mu, from calculations of the scattering (S) parameters for finite-thickness samples. S-parameter retrieval methods have recently become the principal means of characterizing artificially structured metamaterials, which, by nature, are inherently inhomogeneous. While the unit cell of a metamaterial can be made considerably smaller than the free space wavelength, there remains a significant variation of the phase across the unit cell at operational frequencies in nearly all metamaterial structures reported to date. In this respect, metamaterials do not rigorously satisfy an effective medium limit and are closer conceptually to photonic crystals. Nevertheless, we show here that a modification of the standard S-parameter retrieval procedure yields physically reasonable values for the retrieved electromagnetic parameters, even when there is significant inhomogeneity within the unit cell of the structure. We thus distinguish a metamaterial regime, as opposed to the effective medium or photonic crystal regimes, in which a refractive index can be rigorously established but where the wave impedance can only be approximately defined. We present numerical simulations on typical metamaterial structures to illustrate the modified retrieval algorithm and the impact on the retrieved material parameters. We find that no changes to the standard retrieval procedures are necessary when the inhomogeneous unit cell is symmetric along the propagation axis; however, when the unit cell does not possess this symmetry, a modified procedure--in which a periodic structure is assumed--is required to obtain meaningful electromagnetic material parameters. PMID- 15903616 TI - Coherent-mode representation of a statistically homogeneous and isotropic electromagnetic field in spherical volume. AB - It is known that statistically stationary, homogeneous, and isotropic source distributions generate, in an unbounded low-loss medium, an electromagnetic field whose electric cross-spectral density tensor is proportional to the imaginary part of the infinite-space Green tensor. Using the recently established electromagnetic theory of coherent modes, we construct, in a finite spherical volume, the coherent-mode representation of the random electromagnetic field having this property. The analysis covers the fundamental case of blackbody radiation but is valid more generally; since a thermal equilibrium condition is not invoked, the electromagnetic field may have any spectral distribution. Within the scalar theory of coherent modes, which has been available for more than two decades, the analogous formulation results in the first explicit three dimensional coherent-mode representation. PMID- 15903617 TI - Double parametric resonance for matter-wave solitons in a time-modulated trap. AB - We analyze the motion of solitons in a self-attractive Bose-Einstein condensate, loaded into a quasi-one-dimensional parabolic potential trap, which is subjected to time-periodic modulation with an amplitude epsilon and frequency Omega. First, we apply the variational approximation, which gives rise to decoupled equations of motion for the center-of-mass coordinate of the soliton, xi (t), and its width a (t). The equation for xi (t) is the ordinary Mathieu equation (ME) (it is an exact equation that does not depend on the adopted ansatz), the equation for a (t) being a nonlinear generalization of the ME. Both equations give rise to the same map of instability zones in the (epsilon,Omega) plane, generated by the parametric resonances (PRs), if the instability is defined as the onset of growth of the amplitude of the parametrically driven oscillations. In this sense, the double PR is predicted. Direct simulations of the underlying Gross-Pitaevskii equation give rise to a qualitatively similar but quantitatively different stability map for oscillations of the soliton's width a (t). In the direct simulations, we identify the soliton dynamics as unstable if the instability (again, realized as indefinite growth of the amplitude of oscillations) can be detected during a time comparable with, or smaller than, the lifetime of the condensate (therefore accessible to experimental detection). Two-soliton configurations are also investigated. It is concluded that multiple collisions between solitons are elastic, and they do not affect the instability borders. PMID- 15903618 TI - Traceless multipole moment densities and transformations in macroscopic electromagnetism. AB - We consider whether use of traceless multipole moment densities in macroscopic electromagnetism can yield physically acceptable results. For harmonic plane wave fields it is shown that a traceless electric quadrupole density yields linear constitutive relations for which the dynamical material constants (permittivity and magnetoelectric coefficients) and response fields are unphysical. We further show that, within multipole theory, these constitutive relations cannot be transformed into physically acceptable relations. Specifically, the transformed response field D is unphysical for all orders beyond the electric dipole. This contrasts with use of primitive (traced) moment densities, for which unphysical constitutive relations have been successfully transformed up to electric octopole magnetic quadrupole order, thereby providing also the leading contribution to the ac permeability. PMID- 15903619 TI - Oscillations of two-dimensional solitons in harmonic and Bessel optical lattices. AB - We consider parametric amplification of two-dimensional spatial soliton oscillations in longitudinally modulated harmonic and Bessel lattices in Kerr type saturable medium. We show that soliton center oscillations along different axes in two-dimensional lattices are coupled, which gives rise to a number of interesting propagation scenarios including periodic damping and excitation of soliton oscillations along perpendicular axes, selective amplification of soliton oscillations along one transverse axis, and enhancement of soliton spiraling. PMID- 15903620 TI - Influence of impurities on hydrodynamic solitons. AB - The interaction between impurities and solitary waves has been experimentally observed on the surface of a defective water layer subjected to vertical vibration. A slightly rugged surface on one sidewall of the water layer serves as the impurity, making the layer breadth at the defect slightly different elsewhere. A wide-breadth impurity will attract or pin not only the hydrodynamic breather at lower driving frequency, but also the kink at higher driving frequency, while a narrow-breadth one will repel them. Using a multiple scale expansion method, a nonlinear Schro dinger equation with an impurity term (NLSI) was derived from the basic hydrodynamic equation. Furthermore, we present numerical calculations that show good agreement between the NLSI-based theoretical model and the experimental results. PMID- 15903621 TI - Conductance of photons in disordered photonic crystals. AB - The conductance of photons in two-dimensional disordered photonic crystals is calculated using an exact multipole-plane wave method that includes all multiple scattering processes. Conductance fluctuations, the universal nature of which has been established for electrons in the diffusive regime, are studied for photons, in both principal polarizations and for varying disorder. Our simulations show that universal conductance fluctuations can be observed in H(||) (TE) polarization for weak and intermediate disorder while, for E(||) (TM) polarization, we show that the conductance variance is essentially independent of sample size but strongly dependent on disorder. The probability distribution of the conductance is also calculated in the diffusive and localized regimes, and also at their transition, for which the distributions for both polarizations are seen to be very similar. PMID- 15903622 TI - Stabilized vortices in layered Kerr media. AB - In this paper, we demonstrate the possibility of stabilizing beams with angular momentum propagating in Kerr media against filamentation and collapse. Very long propagation distances can be achieved by combining the choice of an appropriate layered medium with alternating focusing and defocusing nonlinearities with the presence of an incoherent guiding beam which is itself stabilized in this medium. The applicability of the results to the field of matter waves is also discussed. PMID- 15903623 TI - Bloch oscillations of Bose-Einstein condensates: breakdown and revival. AB - We investigate the dynamics of Bose-Einstein condensates in a tilted one dimensional periodic lattice within the mean-field (Gross-Pitaevskii) description. Unlike in the linear case the Bloch oscillations decay because of nonlinear dephasing. Pronounced revival phenomena are observed. These are analyzed in detail in terms of a simple integrable model constructed by an expansion in Wannier-Stark resonance states. We also briefly discuss the pulsed output of such systems for stronger static fields. PMID- 15903624 TI - Nonlinear Fano resonance and bistable wave transmission. AB - We consider a discrete model that describes a linear chain of particles coupled to a single-site defect with instantaneous Kerr nonlinearity. We show that this model can be regarded as a nonlinear generalization of the familiar Fano-Anderson model and it can generate amplitude-dependent bistable resonant transmission or reflection. We identify these effects as the nonlinear Fano resonance and study its properties for continuous waves and pulses. PMID- 15903625 TI - Exact numerical solutions for dark waves on the discrete nonlinear Schrodinger equation. AB - In this paper we study numerically existence and stability of exact dark waves on the (nonintegrable) discrete nonlinear Schrodinger equation for a finite one dimensional lattice. These are solutions that bifurcate from stationary dark modes with constant background intensity and zero intensity at a site, and whose initial state translates exactly one site each period of the internal oscillations. We show that exact dark waves are characterized by an oscillatory background whose wavelength is closely related with the velocity. Faster dark waves require smaller wavelengths. For slow enough velocity dark waves are linearly stable, but when trying to continue numerically a solution towards higher velocities bifurcations appear, due to rearrangements in the oscillatory tail in order to make possible a decreasing of the wavelength. However, in principle, one might control the stability of an exact dark wave adjusting a phase factor which plays the role of a discreteness parameter. In addition, we also study the regimes of existence and stability for stationary discrete gray modes, which are exact solutions with phase-twisted constant-amplitude background and nonzero minimum intensity. Also such solutions develop envelope oscillations on top of the homogeneous background when continued into moving phase-twisted solutions. PMID- 15903626 TI - Multisoliton complexes moving on a cnoidal wave background. AB - We obtain solutions of coupled nonlinear Schrodinger equations which describe multisoliton complexes moving on a cnoidal-wave background. Our method is based on the Darboux transformation, which uses Sym's solution of the associated linear equation. Solutions are presented in a matrix determinant form, matrix elements of which are expressed in terms of Jacobi's elliptic functions. Some characteristics of multisoliton complexes like widths and amplitudes are explicitly calculated. PMID- 15903627 TI - Multiple-relaxation-time lattice-Boltzmann model for multiphase flow. AB - The lattice-Boltzmann method has shown promise in simulating multiphase flows. However, when using the Bhatnagar-Gross-Krook (BGK) collision operator and polynomial equilibria, numerical stability problems have been shown to occur as the relaxation time is decreased. Some authors have suggested the use of multiple relaxation-time (MRT) models in lieu of the BGK collision operator, which employs a single relaxation time, to enhance numerical stability. In this paper, a MRT lattice-Boltzmann model for multiphase flow is developed and evaluated for accuracy in several test problems including oscillating liquid cylinders and capillary waves. It is shown that the MRT model is able to achieve numerically stable results at lower viscosities relative to the corresponding BGK model. PMID- 15903628 TI - Adomian's decomposition method for eigenvalue problems. AB - We extend the Adomian's decomposition method to work for the general eigenvalue problems, in addition to the existing applications of the method to boundary and initial value problems with nonlinearity. We develop the Hamiltonian inverse iteration method which will provide the ground state eigenvalue and the explicit form eigenfunction within a few iterations. The method for finding the excited states is also proposed. We present a space partition method for the case that the usual way of series expansion failed to converge. PMID- 15903629 TI - Fractal structure of spin clusters and domain walls in the two-dimensional Ising model. AB - The fractal structure of spin clusters and their boundaries in the critical two dimensional Ising model is investigated numerically. The fractal dimensions of these geometrical objects are estimated by means of Monte Carlo simulations on relatively small lattices through standard finite-size scaling. The obtained results are in excellent agreement with theoretical predictions and partly provide significant improvements in precision over existing numerical estimates. PMID- 15903630 TI - Mirror fluid method for numerical simulation of sedimentation of a solid particle in a Newtonian fluid. AB - The mirror fluid method is proposed for simulating solid-fluid two-phase flow. The whole computational domain is modeled as an Eulerian one for the fluid with a Lagrangian subdomain embedded in it. The boundary condition is enforced implicitly on solid-fluid surface segments by mirror relations. Thus, the total flow is solved in the one domain, in which the solid particle region is replaced with the virtual flow as the mirror image of outside flow. The mirror fluid method is implemented to compute the motion of a rigid spherical or elliptic particle in a Newtonian fluid for the purpose of method validation. The control volume formulation with the SIMPLE algorithm incorporated is used to solve the governing equations on a staggered grid in a two-dimensional coordinate system. A number of numerical experiments on falling particles are performed and the computational results are in good agreement with the reported experimental data. PMID- 15903631 TI - Bias in the direct numerical simulation of isotropic turbulence using the lattice Boltzmann method. AB - Direct numerical simulation of homogeneous, isotropic turbulence using the lattice Boltzmann method is revised. Two-point pressure and velocity correlations are studied and analytical results are derived taking into account the dynamics of the lattice Boltzmann equation. Using the parameters of a two-dimensional (D2Q9) and a three-dimensional (D3Q19) model, it is demonstrated that correlation functions obtained from lattice Boltzmann simulations may have systematic errors at large separation distances due to the second-order error terms. PMID- 15903632 TI - Generalized directed loop method for quantum Monte Carlo simulations. AB - Efficient quantum Monte Carlo update schemes called directed loops have recently been proposed, which improve the efficiency of simulations of quantum lattice models. We propose to generalize the detailed balance equations at the local level during the loop construction by accounting for the matrix elements of the operators associated with open world-line segments. Using linear programming techniques to solve the generalized equations, we look for optimal construction schemes for directed loops. This also allows for an extension of the directed loop scheme to general lattice models, such as high-spin or bosonic models. The resulting algorithms are bounce free in larger regions of parameter space than the original directed loop algorithm. The generalized directed loop method is applied to the magnetization process of spin chains in order to compare its efficiency to that of previous directed loop schemes. In contrast to general expectations, we find that minimizing bounces alone does not always lead to more efficient algorithms in terms of autocorrelations of physical observables, because of the nonuniqueness of the bounce-free solutions. We therefore propose different general strategies to further minimize autocorrelations, which can be used as supplementary requirements in any directed loop scheme. We show by calculating autocorrelation times for different observables that such strategies indeed lead to improved efficiency; however, we find that the optimal strategy depends not only on the model parameters but also on the observable of interest. PMID- 15903633 TI - Simulation method to resolve hydrodynamic interactions in colloidal dispersions. AB - A computational method is presented to resolve hydrodynamic interactions acting on solid particles immersed in incompressible host fluids. In this method, boundaries between solid particles and host fluids are replaced with a continuous interface by assuming a smoothed profile. This enabled us to calculate hydrodynamic interactions both efficiently and accurately, without neglecting many-body interactions. The validity of the method was tested by calculating the drag force acting on a single cylindrical rod moving in an incompressible Newtonian fluid. This method was then applied in order to simulate sedimentation process of colloidal dispersions. PMID- 15903634 TI - Multilevel algorithm for quantum-impurity models. AB - A continuous-time path integral quantum Monte Carlo method using the directed loop algorithm is developed to simulate the Anderson single-impurity model in the occupation number basis. Although the method suffers from a sign problem at low temperatures, the new algorithm has many advantages over conventional algorithms. For example, the model can be easily simulated in the Kondo limit without time discretization errors. Furthermore, many observables including the impurity susceptibility and a variety of fermionic observables can be calculated efficiently. Finally the new approach allows us to explore a general technique, called the multilevel algorithm, to solve the sign problem. We find that the multilevel algorithm is able to generate an exponentially large number of configurations with an effort that grows as a polynomial in inverse temperature such that configurations with a positive sign dominate over those with negative signs. Our algorithm can be easily generalized to other multi-impurity problems. PMID- 15903635 TI - Random initial condition in small Barabasi-Albert networks and deviations from the scale-free behavior. AB - Barabasi-Albert networks are constructed by adding nodes via preferential attachment to an initial core of nodes. We study the topology of small scale-free networks as a function of the size and average connectivity of their initial random core. We show that these two parameters may strongly affect the tail of the degree distribution, by consistently leading to broad-scale or single-scale networks. In particular, we argue that the size of the initial network core and its density of connections may be the main responsible for the exponential truncation of the power-law behavior observed in some small scale-free networks. PMID- 15903636 TI - Vibrational modes and spectrum of oscillators on a scale-free network. AB - We study vibrational modes and spectrum of a model system of atoms and springs on a scale-free network where we assume that the atoms and springs are distributed as nodes and links of a scale-free network. To understand the nature of excitations with many degrees of freedom on the scale-free network, we adopt a particular model that we assign the mass M(i) and the specific oscillation frequency omega(i) of the ith atom and the spring constant K(ij) between the ith and jth atoms. We show that the density of states of the spectrum follows a scaling law P (omega(2)) proportional, variant (omega(2))(-gamma), where gamma = 3 and that as the number of nodes N is increasing, the maximum eigenvalue grows as fast as sqrt[N]. PMID- 15903637 TI - Spatial prisoner's dilemma game with volunteering in Newman-Watts small-world networks. AB - A modified spatial prisoner's dilemma game with voluntary participation in Newman Watts small-world networks is studied. Some reasonable ingredients are introduced to the game evolutionary dynamics: each agent in the network is a pure strategist and can only take one of three strategies (cooperator, defector, and loner); its strategical transformation is associated with both the number of strategical states and the magnitude of average profits, which are adopted and acquired by its coplayers in the previous round of play; a stochastic strategy mutation is applied when it gets into the trouble of local commons that the agent and its neighbors are in the same state and get the same average payoffs. In the case of very low temptation to defect, it is found that agents are willing to participate in the game in typical small-world region and intensive collective oscillations arise in more random region. PMID- 15903638 TI - Universality of rescaled curvature distributions. AB - We applied a recently proposed rescaling of curvatures of eigenvalues of parameter-dependent random matrices to experimental data from acoustic systems and to a theoretical result. It is found that the data from four different experiments, ranging from isotropic plates to anisotropic three-dimensional objects, and the theoretical result always agree with the universal curvature distribution, if only the curvatures are rescaled such that the average of their absolute values is unity. PMID- 15903639 TI - Uniform semiclassical approach to fidelity decay in the deep Lyapunov regime. AB - We use the uniform semiclassical approximation in order to derive the fidelity decay in the regime of large perturbations. Numerical computations are presented which agree with our theoretical predictions. Moreover, our theory allows us to explain previous findings, such as the deviation from the Lyapunov decay rate in cases where the classical finite-time instability is nonuniform in phase space. PMID- 15903640 TI - Simple adaptive-feedback controller for identical chaos synchronization. AB - Based on the invariance principle of differential equations, a simple adaptive feedback scheme is proposed to strictly synchronize almost all chaotic systems. Unlike the usual linear feedback, the variable feedback strength is automatically adapted to completely synchronize two almost arbitrary identical chaotic systems, so this scheme is analytical, and simple to implement in practice. Moreover, it is quite robust against the effect of noise. The famous Lorenz and Rossler hyperchaos systems are used as illustrative examples. PMID- 15903641 TI - Nearest neighbor embedding with different time delays. AB - A nearest neighbor based selection of time delays for phase space reconstruction is proposed and compared to the standard use of time delayed mutual information. The possibility of using different time delays for consecutive dimensions is considered. A case study of numerically generated solutions of the Lorenz system is used for illustration. The effect of contamination with various levels of additive Gaussian white noise is discussed. PMID- 15903642 TI - Galilean invariance and vertex renormalization in turbulence theory. AB - The Navier-Stokes equation is invariant under Galilean transformation of the instantaneous velocity field. However, the total velocity transformation is effected by transformation of the mean velocity alone. For a constant mean velocity, the equation of motion for the fluctuating velocity is automatically Galilean invariant in the comoving frame, and vertex renormalization is not constrained by this symmetry. PMID- 15903643 TI - Ionization fronts in negative corona discharges. AB - We use a hydrodynamic minimal streamer model to study negative corona discharge. By reformulating the model in terms of a quantity called a shielding factor, we deduce laws for the evolution in time of both the radius and intensity of the ionization fronts. We also compute the evolution of the front thickness under the conditions for which it diffuses due to the geometry of the problem and show its self-similar character. PMID- 15903644 TI - Subwavelength far-field resolution in a square two-dimensional photonic crystal. AB - It has been suggested that a subwavelength image can be realized by a photonic crystal slab with a square lattice in the lowest band. However, due to the anisotropy of dispersion in such a photonic structure, the image only appears in the near-field region. In this paper, we show that subwavelength far-field focusing and imaging, explicitly following the well-known wave-beam negative refraction law with relative refractive index of -1, can also be realized by a square photonic crystal through choosing suitable scatterers for the lowest valence band. PMID- 15903645 TI - Bistable diode action in left-handed periodic structures. AB - We study nonlinear transmission of an asymmetric multilayer structure created by alternating slabs of two materials with positive and negative refractive indices. We demonstrate that such a structure exhibits passive spatially nonreciprocal transmission of electromagnetic waves, the analog of the electronic diode. We study the properties of this left-handed diode and confirm its highly nonreciprocal and bistable transmittance by employing direct simulations. PMID- 15903646 TI - Polarization effects in the diffraction of light by a planar chiral structure. AB - We analyze polarization changes of light diffracted on a planar chiral array from the standpoint of the Lorentz reciprocity lemma and find biorthogonality in the polarization eigenstates for waves diffracting though the grating in the opposite direction. Both reciprocal and nonreciprocal components in the polarization azimuth rotation of the diffracted light are identified. The structural chirality of the array arrangement and the chirality of individual elements of the array give rise to polarization effects. PMID- 15903647 TI - Effective elastic parameters of the two-dimensional phononic crystal. AB - The effective acoustic velocity along two highly symmetric directions of the first Brillouin zone is studied in a two-dimensional squarely arranged solid solid phononic crystal. The relationship between the effective elastic velocity and the wave frequency of the two lowest bands is presented, which gives the same results as those obtained directly from the band structure. Numerical calculations show that the effective acoustic velocity of the phononic crystal decreases with frequency increasing from zero to the band edge. PMID- 15903648 TI - Comment on "linear instability of magnetic Taylor-Couette flow with Hall effect". AB - In the paper we comment on [Rudiger and Shalybkov (RS), Phys. Rev. E 69, 016303 (2004)], the instability of the Taylor-Couette flow interacting with a homogeneous background field subject to the Hall effect is studied. We correct a falsely generalizing interpretation of results presented there which could be taken to disprove the existence of the Hall-drift-induced magnetic instability described in Rheinhardt and Geppert, Phys. Rev. Lett. 88, 101103 (2002). It is shown that, in contrast to what is suggested by RS, no additional shear flow is necessary to enable such an instability with a nonpotential magnetic background field, whereas for a curl-free one it is. In the latter case, the instabilities found in RS in situations where neither a hydrodynamic nor a magnetorotational instability exists are demonstrated to be most likely magnetic instead of magnetohydrodynamic. Further, some minor inaccuracies are clarified. PMID- 15903649 TI - Nonlinear rupture of thin liquid films on solid surfaces. AB - In this letter we investigate the rupture instability of thin liquid films by means of a bifurcation analysis in the vicinity of the short-scale instability threshold. The rupture time estimate obtained in closed form as a function of the relevant dimensionless groups is in striking agreement with the results of the numerical simulations of the original nonlinear evolution equations. This suggests that the weakly nonlinear theory adequately captures the underlying physics of the instability. When antagonistic (attractive/repulsive) molecular forces are considered, nonlinear saturation of the instability becomes possible. We show that the stability boundaries are determined by the van der Waals potential alone. PMID- 15903650 TI - Density profiles of temperature-sensitive microgel particles. AB - We have performed small angle neutron scattering measurements (SANS) on dilute aqueous dispersions of polymer microgel particles as a function of temperature, T . The microgel particles are spherical crosslinked assemblies of a loose gel network of a poly-N-isopropylacrylamide (NIPAM) polymer. When the temperature is raised beyond a critical temperature, T(lc) approximately 32 degrees C , the polymer becomes more strongly attracted to itself than the solvent, and the microgel particles contract. The measured form factor, F (q) , for dilute suspensions of uniform microgel particles exhibits many peaks that are characteristic of solid polymer nanospheres. The position and amplitude of the peaks as a function of wave number, q , provide insight into the density profile of poly-NIPAM within the microgels. These peaks can be described well over a wide range of temperature by a model of the polymer density within the particles that is constant up to an inner radius, R1 , and decreases linearly to zero at an outer radius, R2 . PMID- 15903651 TI - Phase transitions induced by noise cross-correlations. AB - A general approach for treating the spatially extended stochastic systems with the nonlinear damping and correlations between additive and multiplicative noises is developed. Within the modified cumulant expansion method, we derive an effective Fokker-Planck equation with stationary solutions that describe the character of the ordered state. We find that the fluctuation cross-correlations lead to a symmetry breaking of the distribution function even in the case of zero dimensional system. In a general case, continuous, discontinuous and reentrant noise induced phase transitions take place. It appears that the cross correlations play the role of bias field which can induce a chain of phase transitions of different nature. Within the mean field approach, we give an intuitive explanation of the system behavior by an effective potential of the thermodynamic type. This potential is written in the form of an expansion with coefficients defined by the temperature, intensity of spatial coupling, autocorrelation and cross-correlation times and intensities of both additive and multiplicative noises. PMID- 15903652 TI - Two approaches toward a high-efficiency flashing ratchet. AB - For a flashing ratchet with periodic potentials fluctuating via random shifts of one-half period, a high efficiency is shown to result from two mechanisms. The previously reported one [Yu. A. Makhnovskii, Phys. Rev. E 69, 021102 (2004); V. M. Rozenbaum, JETP Lett. 79, 388 (2004)] is realized in the near-equilibrium region and implies, first, the presence of a high barrier V0 blocking the reverse movement of a Brownian particle and, second, identical, though energy-shifted, portions of the asymmetric flat potential profile on both half periods. We report another mechanism acting far from equilibrium, typical of strongly asymmetric potentials which are shaped identically on both half periods with a large energetic shift DeltaV . The two mechanisms exhibit radically different limiting behavior of the maximum possible efficiency: eta(m) approximately 1-exp (-beta V0 /2) for the former and eta(m) approximately 1-ln (2betaDeltaV) /betaDeltaV for the latter ( beta being the reciprocal temperature in energy units). The flux and the efficiency for a Brownian motor with a piecewise-linear potential are calculated using the transfer matrix method; an exact analytical solution can thus be obtained for an extremely asymmetric sawtooth potential, the simplest example of the second high-efficiency mechanism. As demonstrated, the mechanisms considered are also characteristic of a two-well periodic potential treated in terms of the kinetic approach. PMID- 15903653 TI - Simulating mesoscopic reaction-diffusion systems using the Gillespie algorithm. AB - We examine an application of the Gillespie algorithm to simulating spatially inhomogeneous reaction-diffusion systems in mesoscopic volumes such as cells and microchambers. The method involves discretizing the chamber into elements and modeling the diffusion of chemical species by the movement of molecules between neighboring elements. These transitions are expressed in the form of a set of reactions which are added to the chemical system. The derivation of the rates of these diffusion reactions is by comparison with a finite volume discretization of the heat equation on an unevenly spaced grid. The diffusion coefficient of each species is allowed to be inhomogeneous in space, including discontinuities. The resulting system is solved by the Gillespie algorithm using the fast direct method. We show that in an appropriate limit the method reproduces exact solutions of the heat equation for a purely diffusive system and the nonlinear reaction-rate equation describing the cubic autocatalytic reaction. PMID- 15903654 TI - Model system for classical fluids out of equilibrium. AB - A model system for classical fluids out of equilibrium, referred to as a dissipative particles dynamics (DPD) solid, is studied by analytical and simulation methods. The time evolution of a DPD particle is described by a fluctuating heat equation. This DPD solid with transport based on collisional transfer (high-density mechanism) is complementary to the Lorentz gas with only kinetic transport (low-density mechanism). Combination of both models covers the qualitative behavior of transport properties of classical fluids over the full density range. The heat diffusivity is calculated using a mean-field theory, leading to a linear-density dependence of this transport coefficient, which is exact at high densities. Subleading density corrections are obtained as well. At lower densities the model has a conductivity threshold below which heat conduction is absent. The observed threshold is explained in terms of percolation diffusion on a random proximity network. The geometrical structure of this network is the same as in continuum percolation of completely overlapping spheres, but the dynamics on this network differs from continuum percolation diffusion. Furthermore, the kinetic theory for DPD is extended to the generalized hydrodynamic regime, where the wave-number-dependent decay rates of the Fourier modes of the energy and temperature fields are calculated. PMID- 15903655 TI - Measuring subdiffusion parameters. AB - We propose a method to extract from experimental data the subdiffusion parameter alpha and subdiffusion coefficient D(alpha) which are defined by means of the relation x(2) = [2 D(alpha) /Gamma (1+alpha) ] t(alpha) where x(2) denotes the mean-square displacement of a random walker starting from x=0 at the initial time t=0 . The method exploits a membrane system where a substance of interest is transported in a solvent from one vessel to another across a thin membrane which plays here only an auxiliary role. Using such a system, we experimentally study the diffusion of glucose and sucrose in a gel solvent. We find a fully analytic solution of the fractional subdiffusion equation with the initial and boundary conditions representing the system under study. Confronting the experimental data with the derived formulas, we show the subdiffusive character of sugar transport in a gel solvent. We precisely determine the parameter alpha , which is smaller than 1, and the subdiffusion coefficient D(alpha) . PMID- 15903656 TI - Role of a finite exposure time on measuring an elastic modulus using microrheology. AB - The role of a finite exposure time sigma on measuring rheological properties using microrheology techniques is theoretically investigated. We concentrate on studying fluid models displaying a plateau in the mean-squared displacement (MSD) of the embedded probe particle. A model is developed to compare the resulting experimentally measured MSD of the particle to its expected value in the fluid model. A plateau MSD is greatly modified in a measurement when sigma is greater than the plateau onset time. Moreover, apparent dynamics drastically differ from the true dynamics at frequencies omega less than or approximately equal sigma(-1) . These results quantify when and how a finite exposure time effects the measured MSD of a probe particle which can then alter the extracted rheological properties and physical interpretations. PMID- 15903657 TI - Noise-induced quantum transport. AB - We analyze the problem of directed quantum transport induced by external exponentially correlated telegraphic noise. In addition to quantum nature of the heat bath, nonlinearity of the periodic system potential brings in quantum contribution. We observe that quantization, in general, enhances classical current at low temperature, while the differences become insignificant at higher temperature. Interplay of quantum diffusion and quantum correction to system potential is analyzed for various ranges of temperature, correlation time and strength of external noise and asymmetry parameters. A possible experimental realization of the observed quantum effects in a superionic conductor placed in a random asymmetric dichotomous electric field has been suggested. PMID- 15903658 TI - Temperature and density relaxation close to the liquid-gas critical point: an analytical solution for cylindrical cells. AB - We present a study of the temperature and density equilibration near the liquid gas critical point of a composite system consisting of a thin circular disk of near-critical fluid surrounded by a copper wall. This system is a simplified model for a proposed space experiment cell that would have 60 thin fluid layers separated by perforated copper plates to aid in equilibration. Upper and lower relaxation time limits that are based on radial and transverse diffusion through the fluid thickness are shown to be too significantly different for a reasonable estimate of the time required for the space experiment. We therefore have developed the first rigorous analytical solution of the piston effect in two dimensions for a cylindrically symmetric three-dimensional cell, including the finite conductivity of the copper wall. This solution covers the entire time evolution of the system after a boundary temperature step, from the early piston effect through the final diffusive equilibration. The calculation uses a quasistatic approximation for the copper and a Laplace-transform solution to the piston effect equation in the fluid. Laplace inversion is performed numerically. The results not only show that the equilibration is divided into three temporal regimes but also give an estimate of the amplitudes of the remaining temperature and density inhomogeneity in each regime. These results yield characteristic length scales for each of the regimes that are used to estimate the expected relaxation times in the one- and two-phase regions near the critical point. PMID- 15903659 TI - Analytical form of the particle distribution based on the cumulant solution of the elastic Boltzmann transport equation. AB - An analytical expression of the particle distribution based on an analytical cumulant solution of the time-dependent elastic Boltzmann transport equation (BTE) is presented. This expression improves upon the previous second order cumulant solution of the BTE described by a Gaussian distribution in two aspects: (1) separating the ballistic component from the scattered component to ensure that the summation in expressions is convergent; and (2) enforcing the causality condition to ensure that no particle travels faster than the free speed of the particles. Time-resolved profiles obtained using the analytical form are compared with those obtained by the Monte Carlo simulation, for both transmission and backscattering. The calculating time using our analytical form is much faster than that using the Monte Carlo approach. PMID- 15903660 TI - Two-dimensional nonlinear nonequilibrium kinetic theory under steady heat conduction. AB - The two-dimensional steady-state Boltzmann equation for hard-disk molecules in the presence of a temperature gradient has been solved explicitly to second order in density and the temperature gradient. The two-dimensional equation of state and some physical quantities are calculated from it and compared with those for the two-dimensional steady-state Bhatnagar-Gross-Krook equation and information theory. We have found that the same kind of qualitative differences as the three dimensional case among these theories still appear in the two-dimensional case. PMID- 15903661 TI - Quantum simulation of the heat capacity of water. AB - Path integral molecular dynamics simulations have been carried out to evaluate heat capacities of water in gas, liquid, and solid phases. For convergence, a 100 ps simulation run with a large path integral variable (P approximately 128) was required even by using the double-centroid virial heat capacity estimator. For all states, the quantum corrections to the heat capacities are significantly large. The calculated heat capacities for vapor and ice I(h) agreed excellently with experimental data, while that for liquid was less than the experimental value by approximately 20% due to the limit of the SPC/F potential model. PMID- 15903662 TI - Segregation mechanisms in a numerical model of a binary granular mixture. AB - A simple phenomenological numerical model of a binary granular mixture is developed and investigated numerically. We attempt to model a recently reported experimental system where a horizontally vibrated binary monolayer was found to exhibit a transition from a mixed to a segregated state as the filling fraction of the mixture was increased. This numerical model is found to reproduce much of the experimentally observed behavior, most importantly the transition from the mixed to the segregated state. We use the numerical model to investigate granular segregation mechanisms and explain the experimentally observed behavior. PMID- 15903663 TI - Small-number statistics near the clustering transition in a compartmentalized granular gas. AB - Statistical fluctuations are observed to profoundly influence the clustering behavior of granular material in a vibrated system consisting of two connected compartments. When the number of particles N is sufficiently large ( N approximately 300 is sufficient), the clustering follows the lines of a standard second-order phase transition and a mean-field description works. For smaller N , however, the enhanced influence of statistical fluctuations breaks the mean-field behavior. We quantitatively describe the competition between fluctuations and mean-field behavior (as a function of N ) using a dynamical flux model and molecular dynamics simulations. PMID- 15903664 TI - Evolution of enduring contacts and stress relaxation in a dense granular medium. AB - Particle contacts in a granular material are formed at different times and have different contact ages, the differences between current time and the times when the contacts were formed. The probability distribution of the contact ages is one of the important statistical properties of particle interactions. The rate of the probability relaxation is proved to be closely related to the stress evolution in the dense granular system. While all particle contacts contribute to the stress, the major contribution is from the contacts with long contact ages compared to the binary collision time of the particles in a dense and slow granular flow, in which particle inertia can be neglected. There is a spectrum of relaxation times in the probability distribution of contact ages. These relaxation times result in different time scales of stress relaxation. As an example, the relations among stress, strain, and the strain rate are studied for a dense granular material undergoing an oscillatory simple shear. The interaction of the time scales determines the fluidlike or solidlike behavior of the material. PMID- 15903665 TI - Short- and long-range topological correlations in two-dimensional aggregation of dense colloidal suspensions. AB - We have studied the average properties and the topological correlations of computer-simulated two-dimensional (2D) aggregating systems at different initial surface packing fractions. For this purpose, the centers of mass of the growing clusters have been used to build the Voronoi diagram, where each cell represents a single cluster. The number of sides (n) and the area (A) of the cells are related to the size of the clusters and the number of nearest neighbors, respectively. We have focused our paper in the study of the topological quantities derived from number of sides, n , and we leave for a future work the study of the dependence of these magnitudes on the area of the cells, A . In this work, we go beyond the adjacent cluster correlations and explore the organization of the whole system of clusters by dividing the space in concentric layers around each cluster: the shell structure. This method allows us to analyze the time behavior of the long-range intercluster correlations induced by the aggregation process. We observed that kinetic and topological properties are intimately connected. Particularly, we found a continuous ordering of the shell structure from the earlier stages of the aggregation process, where clusters positions approach a hexagonal distribution in the plane. For long aggregation times, when the dynamic scaling regime is achieved, the short- and long-range topological properties reached a final stationary state. This ordering is stronger for high particle densities. Comparison between simulation and theoretical data points out the fact that 2D colloidal aggregation in the absence of interactions (diffusion limited cluster aggregation regimen) is only able to produce short-range cluster cluster correlations. Moreover, we showed that the correlation between adjacent clusters verifies the Aboav-Weaire law, while all the topological properties for nonadjacent clusters are mainly determined by only two parameters: the second central moment of number-of-sides distribution mu(2) = sumP (n) (n-6)(2) and the screening factor a (defined through the Aboav-Weaire equation). We also found that the values of mu(2) and a calculated for two-dimensional aggregating system are related through a single universal common form a proportional to mu2(-0.89), which is independent of the particle concentration. PMID- 15903666 TI - Depletion interaction in a quasi-two-dimensional colloid assembly. AB - We address several aspects of the character of the depletion interaction in a quasi-two-dimensional (Q2D) colloid system. First, we consider how, given information concerning the pair and triplet correlation functions, the depletion interaction can be efficiently and accurately determined. For this purpose we introduce a method based on the Born-Green equation using the assumption that for the Q2D binary mixtures of interest to us the depletion interaction is accurately represented as a sum of pair potentials. We then verify, by direct calculation, that three-particle contributions to the depletion interaction are negligibly small in the region of thermodynamic state space of interest to us. Second, we develop a representation of the dependence of the depletion interaction in a Q2D colloid system on the thickness of the confining parallel plates. Third, we report the results of extensive simulations of Q2D binary hard-sphere mixtures for a range of cell thickness, large and small particle number densities, and a ratio of sphere diameters q=0.3 . PMID- 15903667 TI - Influence of polydispersity on the effective interaction in a quasi-two dimensional pseudo-one-component colloid fluid. AB - We report the results of simulations of one-component and binary quasi-two dimensional polydisperse hard sphere colloid fluids. When the polydisperse one component system is regarded as an effective monodisperse one-component system, our results show that polydispersity gives rise to effective pair attraction and soft repulsion. These effective interactions depend on both the degree of polydispersity and the system density. At low density only the effective soft repulsion survives and the effective attraction becomes negligible. For the binary system, the depletion interaction develops extra features due to polydispersity, specifically a repulsive interaction with range twice the average particle diameter. At low density this extra feature vanishes. To carry out our study we have devised a method for continuous sampling of the polydisperse mixture according to a prescribed particle diameter distribution. PMID- 15903668 TI - Variational bounds for the relaxation times of swelling gels. AB - Variational bounds are found for the relaxation times of a gel of general shape swelling in a solvent based on the stress-diffusion coupling model. It is shown that in the case of free swelling, the longest relaxation time is inversely proportional to the osmotic modulus K in the limit of K-->0 and K-->infinity . This indicates that the relaxation time diverges at the point of K=0 . The divergence, however, disappears if a part of the gel is mechanically constrained. PMID- 15903669 TI - Barrier hopping, viscous flow, and kinetic gelation in particle-polymer suspensions. AB - The naive mode coupling-polymer reference interaction site model (MCT-PRISM) theory of gelation and elasticity of suspensions of hard sphere colloids or nanoparticles mixed with nonadsorbing polymers has been extended to treat the emergence of barriers, activated transport, and viscous flow. The barrier makes the dominant contribution to the single particle relaxation time and shear viscosity, and is a rich function of the depletion attraction strength via the polymer concentration, polymer-particle size asymmetry ratio, and particle volume fraction. The dependences of the barrier on these three system parameters can be accurately collapsed onto a single scaling variable, and the resultant master curve is well described by a power law. Nearly universal master curves are also constructed for the hopping or alpha relaxation time for system conditions not too close to the ideal MCT transition. Based on the calculated barrier hopping time, a theory for kinetic gel boundaries is proposed. The form and dependence on system parameters of the kinetic gel lines are qualitatively the same as obtained from prior ideal MCT-PRISM studies. The possible relevance of our results to the phenomenon of gravity-driven gel collapse is studied. The general approach can be extended to treat nonlinear viscoelasticity and rheology of polymer-colloid suspensions and gels. PMID- 15903670 TI - Crystallization in settling mixtures of colloidal spheres and plates. AB - The behavior of mixtures of silica spheres and smaller silica-coated gibbsite plates is studied by means of confocal microscopy. Addition of plates slows down the crystallization of the settling spheres. Liquidlike microphases of the plates are found in the sediments of the spheres. It is argued that this is due to simultaneous sedimentation of the plates and spheres as well as depletion interaction between both species. Typical length scales in the sediments, derived from Fourier transforms of the confocal images, suggest there are still specific interactions present. PMID- 15903671 TI - Electro-osmotic flow of a model electrolyte. AB - Electro-osmotic flow is studied by nonequilibrium molecular dynamics simulations in a model system chosen to elucidate various factors affecting the velocity profile and facilitate comparison with existing continuum theories. The model system consists of spherical ions and solvent, with stationary, uniformly charged walls that make a channel with a height of 20 particle diameters. We find that hydrodynamic theory adequately describes simple pressure-driven (Poiseuille) flow in this model. However, Poisson-Boltzmann theory fails to describe the ion distribution in important situations, and therefore continuum fluid dynamics based on the Poisson-Boltzmann ion distribution disagrees with simulation results in those situations. The failure of Poisson-Boltzmann theory is traced to the exclusion of ions near the channel walls resulting from reduced solvation of the ions in that region. When a corrected ion distribution is used as input for hydrodynamic theory, agreement with numerical simulations is restored. An analytic theory is presented that demonstrates that repulsion of the ions from the channel walls increases the flow rate, and attraction to the walls has the opposite effect. A recent numerical study of electro-osmotic flow is reanalyzed in the light of our findings, and the results conform well to our conclusions for the model system. PMID- 15903672 TI - Application of the Bethe-Peierls approximation to a lattice-gas model of adsorption on mesoporous materials. AB - We calculate adsorption and desorption isotherms in models of several classes of porous materials using a lattice-gas model solved in the Bethe-Peierls (quasichemical) approximation. Isotherms and fluid density profiles from the Bethe-Peierls and Bragg-Williams approximations are compared with grand-canonical Monte Carlo simulation results. The Bethe-Peierls approximation produces both more accurate adsorption and desorption isotherms and more realistic fluid density profiles than the Bragg-Williams approximation. Details of the application of the Bethe-Peierls approximation applied to a three-dimensionally inhomogeneous system are given. We show that the numerical solution of this theory can be accomplished using a self-consistent iterator very similar to that currently used in studies employing the Bragg-Williams approximation. This iterative scheme is substantially more efficient than the numerical optimization method used in many previous studies of lattice-gas models in the quasichemical approximation. We find that use of the Bethe-Peierls approximation is only slightly more computationally demanding than the Bragg-Williams approximation, and thus recommend it for use in future work on this class of models. PMID- 15903673 TI - Viscosity anomaly near the critical point in nitrobenzene + alkane binary systems. AB - The viscosity near the critical point in nitrobenzene+hexane and nitrobenzene+heptane binary systems was studied by examining the viscosity values for critical mixtures at a variable temperature as obtained with a falling-ball viscometer. The regular part of the viscosity of the critical mixtures was calculated by interpolating measurements made at noncritical concentrations. Because viscosity anomaly studies must be conducted at zero shear, a method allowing the estimation of the effective shear for this type of viscometer was developed with a view to introducing the corrections required. This methodology was used to determine the critical exponent for the viscosity anomaly in nitrobenzene+hexane and nitrobenzene+heptane systems, which were found to be 0.0422+/-0.0004 and 0.0432+/-0.0013 , respectively, very consistent with the accepted value: 0.043. PMID- 15903674 TI - Finite-size effects and intermittency in a simple aging system. AB - We study the intermittent dynamics and the fluctuations of the dynamic correlation function of a simple aging system. Given its size L and its coherence length xi , the system can be divided into N independent subsystems, where N= (L/xi)(d), and d is the dimension of space. Each of them is considered as an aging subsystem which evolves according to an activated dynamics between energy levels. We compute analytically the distribution of trapping times for the global system, which can take power-law, stretched-exponential or exponential forms according to the values of N and the regime of times considered. An effective number of subsystems at age t(w), N(eff) (t(w)), can be defined, which decreases as t(w) increases, as well as an effective coherence length, xi (t(w)) approximately tw ((1-mu)/d), where mu<1 characterizes the trapping times distribution of a single subsystem. We also compute the probability distribution functions of the time intervals between large decorrelations, which exhibit different power-law behaviors as t(w) increases (or N decreases), and which should be accessible experimentally. Finally, we calculate the probability distribution function of the two-time correlator. We show that in a phenomenological approach, where N is replaced by the effective number of subsystems N(eff) (t(w)), the same qualitative behaviour as in experiments and simulations of several glassy systems can be obtained. PMID- 15903675 TI - Dynamical susceptibility of glass formers: contrasting the predictions of theoretical scenarios. AB - We compute analytically and numerically the four-point correlation function that characterizes nontrivial cooperative dynamics in glassy systems within several models of glasses: elastoplastic deformations, mode-coupling theory (MCT), collectively rearranging regions (CRR's), diffusing defects, and kinetically constrained models (KCM's). Some features of the four-point susceptibility chi(4) (t) are expected to be universal: at short times we expect a power-law increase in time as t(4) due to ballistic motion (t(2) if the dynamics is Brownian) followed by an elastic regime (most relevant deep in the glass phase) characterized by a t or sqrt[t] growth, depending on whether phonons are propagative or diffusive. We find in both the beta and early alpha regime that chi(4) approximately t(mu), where mu is directly related to the mechanism responsible for relaxation. This regime ends when a maximum of chi(4) is reached at a time t= t(*) of the order of the relaxation time of the system. This maximum is followed by a fast decay to zero at large times. The height of the maximum also follows a power law chi(4) (t(*)) approximately t(*lambda). The value of the exponents mu and lambda allows one to distinguish between different mechanisms. For example, freely diffusing defects in d=3 lead to mu=2 and lambda=1 , whereas the CRR scenario rather predicts either mu=1 or a logarithmic behavior depending on the nature of the nucleation events and a logarithmic behavior of chi(4) (t(*)) . MCT leads to mu=b and lambda=1/gamma , where b and gamma are the standard MCT exponents. We compare our theoretical results with numerical simulations on a Lennard-Jones and a soft-sphere system. Within the limited time scales accessible to numerical simulations, we find that the exponent mu is rather small, mu<1 , with a value in reasonable agreement with the MCT predictions, but not with the prediction of simple diffusive defect models, KCM's with noncooperative defects, and CRR's. Experimental and numerical determination of chi(4) (t) for longer time scales and lower temperatures would yield highly valuable information on the glass formation mechanism. PMID- 15903676 TI - Kinetic self-assembly of metals on copolymer templates. AB - In this work, we seek to understand some of the fundamental processes that govern self-assembly at the nanoscale in the context of the formation of metallic structures on patterned copolymer templates. To this end, we focus on recent experiments where morphologies resulting from the evaporation-deposition of different metals on PS- b -PMMA phase-separated templates were studied. We show that the different morphologies obtained can be understood in terms of the relative importance of the energetics and kinetics. We then focus on a particular morphology: micron-long wirelike states obtained by the evaporation deposition of silver on the template. We show the existence of "nontrivial" correlations between adjacent wires that can be understood based on a purely kinetic mechanism. We also compare these correlations quantitatively to those obtained from simulations done with the relevant experimental parameters and find them in good agreement. PMID- 15903677 TI - Phase change of a confined subcooled simple liquid in a nanoscale cavity. AB - The phase transition of a simple liquid bounded between two parallel walls a few nanometers apart is investigated with molecular dynamics simulations. Vapor nucleation in a liquid confined in a microchannel of only a few nanometers in size cannot be achieved by increasing the temperature at the wall. Already small changes in temperature cause a large rise in pressure, in terms of orders of magnitude. On the other hand, using the fact that some fluids thermally contract on cooling, e.g., the argon liquid investigated here, reducing the temperature places the fluid in the liquid-vapor coexistence regime. If the bottom wall temperature is further reduced, the fluid will crystallize starting from the bottom surface, creating a "frozen" bubble in the crystallized state. It was found that the confining walls and not the quenching rate primarily affect the crystallization process. However, the fastest cooling rate investigated herein led to a decrease of the boiling initiation temperature. At a lower cooling rate, the vapor nucleation temperature was the same as the equilibrium boiling temperature for the confined liquid. Small temperatures in the confined system result in dominating attraction forces at the fluid-wall interface exposing the fluid to tensile stress. The increased influence of the walls results in a significant decrease of the boiling as well as freezing temperatures. PMID- 15903678 TI - Irreversible aggregation of interacting particles in one dimension. AB - We present a study of the aggregation of interacting particles in one dimension. This situation, for example, applies to atoms trapped along linear defects at the surface of a crystal. Simulations are performed with two lattice models. In the first model, the borders of atoms and islands interact in a vectorial manner via force monopoles. In the second model, each atom carries a dipole. These two models lead to qualitatively similar but quantitatively different behaviors. In both cases, the final average island size S(f) does not depend on the interactions in the limits of very low and very high coverages. For intermediate coverages, S(f) exhibits an asymmetric behavior as a function of the interaction strength: while it saturates for attractive interactions, it decreases for repulsive interactions. A class of mean-field models is designed, which allows one to retrieve the interaction dependence on the coverage dependence of the average island size with a good accuracy. PMID- 15903679 TI - Dramatic enhancement of capillary wave fluctuations of a decorated water surface. AB - We have demonstrated by x-ray diffuse scattering that a bimolecular layer of a preformed three-tailed amphiphile, ferric stearate, drastically enhances capillary wave fluctuations on water surface due to a reduction in surface tension to 1 mN/m . The bimolecular layer is composed of molecules in symmetric configuration, on top of molecules in asymmetric configuration with ferric ions in contact with water. Unlike the usual Langmuir monolayers, this layer of molecules does not rupture under compression, but becomes thicker. This behavior mimics folding of a membrane on a liquid surface and is closely related to the cohesive interaction brought by the ferric ions. The low effective tension of this artificial membrane depends on the available area and reduces as the microscopic excess area increases. PMID- 15903680 TI - Kinetics of step bunching during growth: a minimal model. AB - We study a minimal stochastic model of step bunching during growth on a one dimensional vicinal surface. The formation of bunches is controlled by the preferential attachment of atoms to descending steps (inverse Ehrlich-Schwoebel effect) and the ratio d of the attachment rate to the terrace diffusion coefficient. For generic parameters (d>0) the model exhibits a very slow crossover to a nontrivial asymptotic coarsening exponent beta approximately 0.38 . In the limit of infinitely fast terrace diffusion (d=0) linear coarsening (beta=1) is observed instead. The different coarsening behaviors are related to the fact that bunches attain a finite speed in the limit of large size when d=0 , whereas the speed vanishes with increasing size when d>0 . For d=0 an analytic description of the speed and profile of stationary bunches is developed, and a connection to the problem of front propagation into an unstable state is pointed out. PMID- 15903681 TI - Polynuclear growth model with external source and random matrix model with deterministic source. AB - We present a random matrix interpretation of the distribution functions which have appeared in the study of the one-dimensional polynuclear growth (PNG) model with external sources. It is shown that the distribution, GOE2 , which is defined as the square of the Gaussian orthogonal ensemble (GOE) Tracy-Widom distribution, can be obtained as the scaled largest eigenvalue distribution of a special case of a random matrix model with a deterministic source, which have been studied in a different context previously. Compared to the original interpretation of the GOE2 as "the square of GOE," ours has an advantage in that it can also describe the transition from the Gaussian unitary ensemble (GUE) Tracy-Widom distribution to the GOE2 . We further demonstrate that our random matrix interpretation can be obtained naturally by noting the similarity of the topology between a certain noncolliding Brownian motion model and the multilayer PNG model with an external source. This provides us with a multimatrix model interpretation of the multipoint height distributions of the PNG model with an external source. PMID- 15903682 TI - Modeling phase transitions during the crystallization of a multicomponent fat under shear. AB - The crystallization of multicomponent systems involves several competing physicochemical processes that depend on composition, temperature profiles, and shear rates applied. Research on these mechanisms is necessary in order to understand how natural materials form crystalline structures. Palm oil was crystallized in a Couette cell at 17 and 22 degrees C under shear rates ranging from 0 to 2880 s(-1) at a synchrotron beamline. Two-dimensional x-ray diffraction patterns were captured at short time intervals during the crystallization process. Radial analysis of these patterns showed shear-induced acceleration of the phase transition from alpha to beta(') . This effect can be explained by a simple model where the alpha phase nucleates from the melt, a process which occurs independently of shear rate. The alpha phase grows according to an Avrami growth model. The beta(') phase nucleates on the alpha crystallites, with the amount of beta(') crystal formation dependent on the rate of transformation of alpha to beta(') as well as the growth rate of the beta(') phase from the melt. The shear induced alpha- beta(') phase transition acceleration occurs because under shear, the alpha nuclei form many distinct small crystallites which can easily transform to the beta(') form, while at lower shear rates, the alpha nuclei tend to aggregate, thus retarding the nucleation of the beta(') crystals. The displacement of the diffraction peak positions revealed that increased shear rate promotes the crystallization of the higher melting fraction, affecting the composition of the crystallites. Crystalline orientation was observed only at shear rates above 180 s(-1) at 17 degrees C and 720 s(-1) at 22 degrees C . PMID- 15903683 TI - Slip behavior in liquid films on surfaces of patterned wettability: comparison between continuum and molecular dynamics simulations. AB - We investigate the behavior of the slip length in Newtonian liquids subject to planar shear bounded by substrates with mixed boundary conditions. The upper wall, consisting of a homogenous surface of finite or vanishing slip, moves at a constant speed parallel to a lower stationary wall, whose surface is patterned with an array of stripes representing alternating regions of no shear and finite or no slip. Velocity fields and effective slip lengths are computed both from molecular dynamics (MD) simulations and solution of the Stokes equation for flow configurations either parallel or perpendicular to the stripes. Excellent agreement between the hydrodynamic and MD results is obtained when the normalized width of the slip regions, a/sigma greater than or approximately equal O (10) , where sigma is the (fluid) molecular diameter characterizing the Lennard-Jones interaction. In this regime, the effective slip length increases monotonically with a/sigma to a saturation value. For a/sigma less than or approximately O (10) and transverse flow configurations, the nonuniform interaction potential at the lower wall constitutes a rough surface whose molecular scale corrugations strongly reduce the effective slip length below the hydrodynamic results. The translational symmetry for longitudinal flow eliminates the influence of molecular scale roughness; however, the reduced molecular ordering above the wetting regions of finite slip for small values of a/sigma increases the value of the effective slip length far above the hydrodynamic predictions. The strong correlation between the effective slip length and the liquid structure factor representative of the first fluid layer near the patterned wall illustrates the influence of molecular ordering effects on slip in noninertial flows. PMID- 15903684 TI - Multicomponent alloy solidification: phase-field modeling and simulations. AB - A general formulation of phase-field models for nonisothermal solidification in multicomponent and multiphase alloy systems is derived from an entropy functional in a thermodynamically consistent way. General expressions for the free energy densities, for multicomponent diffusion coefficients, and for both weak and faceted types of surface energy and kinetic anisotropy are possible. A three dimensional simulator is developed to show the capability of the model to describe phase transitions, complex microstructure formation, and grain growth in polycrystalline textures. PMID- 15903685 TI - Effect of optical purity on phase sequence in antiferroelectric liquid crystals. AB - We use the discrete phenomenological model to study theoretically the phase diagrams in antiferroelectric liquid crystals (AFLCs) as a function of optical purity and temperature. High sensitivity of the phase sequence in the AFLCs to optical purity is attributed to the piezoelectric coupling which is reduced if optical purity is reduced. We limit our study to three topologically equal smetic (Sm)phases: Sm-C(*), Sm-C(*)(alpha), and Sm-C(*)(A) and show that the reduction of optical purity forces the system from the antiferroelectric to the ferroelectric phase with a possible Sm- C(*)(alpha) between them. The effect of the flexoelectric and the quadratic coupling is considered as well. If the phase diagram includes only two phases, Sm-C* and Sm- C*(A), the flexoelectric coupling is very small. The materials which exhibit the Sm-C(*)(alpha) in a certain range of optical purity and temperature, can be expected to have a significant flexoelectric coupling that is comparable with the piezoelectric coupling. Upon lowering the temperature the phase sequence Sm-A-->Sm-C(*)(alpha)-->Sm-C*-->Sm C(*)(A) is possible in systems where quadratic coupling is very strong. PMID- 15903686 TI - Mechanical actions on nanocylinders in nematic liquid crystals. AB - We apply the Landau-de Gennes theory to study the equilibrium problem that arises when a cylinder of radius R is kept at a given distance h from a plane wall. We assume that both the lateral boundary of the cylinder and the wall enforce homeotropic anchoring conditions on the liquid crystal, which prescribe the liquid crystal molecules to stick orthogonally to the bounding surfaces. Typically, in our study R ranges from a few to hundreds of biaxial coherence lengths, where a biaxial coherence length, which depends on the temperature, is a few nanometers. The equilibrium textures exhibit a bifurcation between a flat solution, where one eigenvector of the order tensor Q is everywhere parallel to the cylinder's axis, and an escape solution, where the eigenframe of Q flips out of the plane orthogonal to the cylinder's axis. The escape texture minimizes an appropriately renormalized energy functional F(*) for h>h(c), while the flat texture minimizes F(*) for h< h(c). We compute both the force and the torque transmitted to the cylinder by the surrounding liquid crystal and we find that the diagrams of both as functions of h fail to be monotonic along the escape texture. Thus, upon decreasing h, a snapping instability is predicted to occur, with an associated hysteresis loop in the force diagram, before h reaches h(c). Finally, since the symmetry of this problem makes it equivalent to the one where two parallel cylinders are separated by the distance 2h , the snapping instability predicted here should also be observed there. PMID- 15903687 TI - Forces in nematic liquid crystals constrained to the nanometer scale under hybrid anchoring conditions. AB - Using a surface forces apparatus we have studied two thermotropic nematic liquid crystals (5CB and ME10.5) subjected to hybrid (homeotropic/planar) anchoring conditions. A film of nematic material is constrained between two curved smooth surfaces separated by less than 2500 A . The intersurface force is nonmonotonic with the separation, being repulsive for thicknesses larger than approximately 100 A and strongly adhesive at a shorter scale. While the repulsion can be qualitatively explained by an elastic model of director deformation, including anchoring deviation at the boundaries, the attraction cannot be explained either by elasticity or by dispersive forces. The expected confinement-induced anchoring transition has not been observed for a thickness as small as 200 A . PMID- 15903688 TI - Dielectric spectroscopy of de Vries-type smectic-A* -smectic-C* transitions. AB - We report results of dielectric investigations on a number of ferroelectric liquid crystalline (FLC) compounds with different degrees of layer shrinkage in the smectic-A(*) -smectic-C(*) phase transitions. With a decreasing extent of layer shrinkage the investigated FLCs exhibit a significantly increasing soft mode absorption, decreasing leading Landau coefficient alpha, and a considerably broader mean-field regime. We explain these tendencies by the fact that the low layer shrinkage materials come closer to the diffuse cone model of de Vries than to the common model of rigid rods which maintain their orientational order during tilting. In the case of the diffuse cone model the tilt-angle fluctuations are decoupled from variations of the smectic layer spacing, which explains the observed behavior. PMID- 15903689 TI - Evidence for de Vries structure in a smectic-A liquid crystal observed by polarized Raman scattering. AB - The second- and fourth-order apparent orientational order parameters of the core part of the molecule P2 (app) and P4 (app) , have been measured by polarized vibrational Raman spectroscopy for a homogeneously aligned ferroelectric smectic liquid crystal with three dimethyl siloxane groups in the achiral terminal chain, which shows de Vries-type phenomena, i.e., very large electroclinic effect in the smectic- A (Sm-A) phase and a negligible layer contraction at the phase transition between the Sm-A and Sm- C(*) phases. The apparent orientational order parameters of the rigid core part of the molecule are extremely small both with and without the external electric field in Sm-A . These results provide evidence for the existence of the de Vries Sm-A phase, where the local molecular director is tilted at a large angle. PMID- 15903690 TI - Stokes parameter studies of spontaneous emission from chiral nematic liquid crystals as a one-dimensional photonic stopband crystal: experiment and theory. AB - The helical structure of uniformly aligned chiral nematic liquid crystals results in a photonic stopband for only one sense of circular polarization. The spectroscopic Stokes polarimeter is used to analyze spontaneous emission in the stopband. Highly polarized photoluminescence is found and the polarization properties vary with the excitation wavelength. Spontaneous emission is enhanced at the stopband edge and this Purcell effect is greater on excitation at wavelengths where the absorption coefficient is low. This is interpreted as greater overlap between the excited molecules and the electrical modal field of the resonant modes at the stopband edge. Photoluminescence detected from the excitation face of the liquid crystal cell is less polarized because of photon tunneling. Fermi's golden rule in conjunction with Stokes vectors is used to model the polarization of emission taking multiple reflections at the interfaces of the cell into account. The discrepancy between the experiment and the theoretical model is interpreted as direct experimental evidence that virtual photons, which originate from zero point fluctuations of quantum space, are randomly polarized. PMID- 15903691 TI - Glass transition in ultrathin polymer films: a thermal expansion study. AB - The glass transition process gets affected in ultrathin films having thickness comparable to the size of the molecules. We observe systematic broadening of the glass transition temperature (T(g)) as the thickness of an ultrathin polymer film reduces below the radius of gyration but the change in the average T(g) was found to be very small. The existence of reversible negative and positive thermal expansion below and above T(g) increased the sensitivity of our thickness measurements performed using energy-dispersive x-ray reflectivity. A simple model of the T(g) variation as a function of depth expected from sliding motion could explain the results. PMID- 15903692 TI - Influence of molecular weight on the phase behavior and structure formation of branched side-chain hairy-rod polyfluorene in bulk phase. AB - We report on an experimental study of the self-organization and phase behavior of hairy-rod pi -conjugated branched side-chain polyfluorene, poly[9,9-bis(2 ethylhexyl)-fluorene-2,7-diyl]-i.e., poly[2,7-(9,9-bis(2-ethylhexyl)fluorene] (PF2/6) -as a function of molecular weight (M(n)) . The results have been compared to those of phenomenological theory. Samples for which M(n) =3-147 kg/mol were used. First, the stiffness of PF2/6 , the assumption of the theory, has been probed by small-angle neutron scattering in solution. Thermogravimetry has been used to show that PF2/6 is thermally stable over the conditions studied. Second, the existence of nematic and hexagonal phases has been phenomenologically identified for lower and higher M(n) (LMW, M(n) < M(*)(n) and HMW, M(n) > M(*)(n) ) regimes, respectively, based on free-energy argument of nematic and hexagonal hairy rods and found to correspond to the experimental x-ray diffraction (XRD) results for PF2/6 . By using the lattice parameters of PF2/6 as an experimental input, the nematic-hexagonal transition has been predicted in the vicinity of glassification temperature (T(g)) of PF2/6 . Then, by taking the orientation parts of the free energies into account the nematic-hexagonal transition has been calculated as a function of temperature and M(n) and a phase diagram has been formed. Below T(g) of 80 degrees C only (frozen) nematic phase is observed for M(n)< M(*)(n) = 10(4) g/mol and crystalline hexagonal phase for M(n) > M(*)(n) . The nematic-hexagonal transition upon heating is observed for the HMW regime depending weakly on M(n) , being at 140-165 degrees C for M(n) > M(*)(n). Third, the phase behavior and structure formation as a function of M(n) have been probed using powder and fiber XRD and differential scanning calorimetry and reasonable semiquantitative agreement with theory has been found for M(n) >or=3 kg/mol. Fourth, structural characteristics are widely discussed. The nematic phase of LMW materials has been observed to be denser than high-temperature nematic phase of HMW compounds. The hexagonal phase has been found to be paracrystalline in the (ab0) plane but a genuine crystal meridionally. We also find that all these materials including the shortest 10-mer possess the formerly observed rigid five helix hairy-rod molecular structure. PMID- 15903693 TI - Aging phenomena in poly(methyl methacrylate) thin films: memory and rejuvenation effects. AB - The aging dynamics in thin films of poly(methyl methacrylate) (PMMA) have been investigated through dielectric measurements for different types of aging processes. The dielectric constant was found to decrease with increasing aging time at an aging temperature in many cases. An increase in the dielectric constant was also observed in the long-time region (>or=11 h) near the glass transition temperature for thin films with thickness less than 26 nm . In the constant-rate mode including a temporary stop at a temperature T(a) , the memory of the aging at T(a) was found to be kept and then to be recalled during the subsequent heating process. In the negative-temperature cycling process, a strong rejuvenation effect has been observed after a temperature shift from the initial temperature T1 to the second temperature T2 (= T1 +DeltaT) when DeltaT approximately -20 K . Furthermore, a full memory effect has also been observed for the temperature shift from T2 to T1 . This suggests that the aging at T1 is totally independent of that at T2 for DeltaT approximately -20 K. As /DeltaT/ decreases, the independence of the aging between the two temperatures was found to be weakened-i.e., the effective time, which is a measure of the contribution of the aging at T1 to that at T2 , is a decreasing function of /DeltaT/ in the negative region of DeltaT . As the film thickness decreases from 514 nm to 26 nm, the /DeltaT/ dependence of the effective time was found to become much stronger. The contribution of the aging at T2 to that at T1 disappears more rapidly with increasing /DeltaT/ in thin-film geometry than in the bulk state. PMID- 15903694 TI - Smectic ordering of homogeneous semiflexible polymers. AB - A self-consistent-field theory for fluids of homogeneous wormlike polymers exhibiting a one-dimensional spatial variation is presented. We have extended the treatment of excluded-volume effects by adding an effective interaction term which describes the excluded volume between wormlike cylindrical segments and terminal (or end) segments of the polymer molecules. This enables us to find a smectic-A phase in the case of homogeneous semiflexible polymers. Using this framework, we have investigated the occurrence of smectic-A, nematic, and isotropic phases in the second-virial (Onsager) approximation. Phase diagrams are calculated for systems characterized by different rigidities (i.e., persistence lengths). For the case of infinitely rigid molecules, the nematic-smectic transition appears to be mostly second order. Systems of semiflexible molecules exhibit mainly a first-order smectic-nematic transition, and their isotropic nematic-smectic triple points are accessed for different rigidity values. The nematic-smectic transition line is in good agreement with previous analytical calculations, which were also performed assuming the second-virial approximation. However, the values of the volume fraction at the nematic-smectic transition are large compared with computer simulation results, indicating limitations of the second-virial approximation. PMID- 15903695 TI - Electrical conduction in macroscopically oriented deoxyribonucleic and hyaluronic acid samples. AB - Measurements of the quasistatic and frequency dependent electrical conductivity below 1 MHz were carried out on wet-spun, macroscopically oriented, calf thymus deoxyribonucleic (DNA) and umbilical cord hyaluronic acid (HA) bulk samples. The frequency dependence of the electrical conductivity in the frequency range of approximately 10(-3) - 10(6) Hz of both materials is surprisingly rather similar. Temperature dependence of the quasistatic electrical conductivity above the low temperature saturation plateau can be well described by the activated Arrhenius law with the activation energy of approximately 0.8 eV for both DNA and HA. We discuss the meaning of these findings for the possible conduction mechanism in these particular charged polyelectrolytes. PMID- 15903696 TI - Modeling the effects of anesthesia on the electroencephalogram. AB - Changes to the electroencephalogram (EEG) observed during general anesthesia are modeled with a physiological mean field theory of electrocortical activity. To this end a parametrization of the postsynaptic impulse response is introduced which takes into account pharmacological effects of anesthetic agents on neuronal ligand-gated ionic channels. Parameter sets for this improved theory are then identified which respect known anatomical constraints and predict mean firing rates and power spectra typically encountered in human subjects. Through parallelized simulations of the eight nonlinear, two-dimensional partial differential equations on a grid representing an entire human cortex, it is demonstrated that linear approximations are sufficient for the prediction of a range of quantitative EEG variables. More than 70,000 plausible parameter sets are finally selected and subjected to a simulated induction with the stereotypical inhaled general anesthetic isoflurane. Thereby 86 parameter sets are identified that exhibit a strong "biphasic" rise in total power, a feature often observed in experiments. A sensitivity study suggests that this "biphasic" behavior is distinguishable even at low agent concentrations. Finally, our results are briefly compared with previous work by other groups and an outlook on future fits to experimental data is provided. PMID- 15903697 TI - Using cell potential energy to model the dynamics of adhesive biological cells. AB - Developing a continuous mathematical model of a physical phenomenon which is based on a discrete model of the same system is not straightforward. Yet such a process is useful in illustrating the link between the individual behavior of the elements comprising a system and its macroscopic behavior. Collections of biological cells can exhibit phenomena such as pattern formation, aggregation, and invasion, and mathematics has proven useful in elucidating the underlying dynamics of these phenomena. The continuous models formulated are frequently of reaction-diffusion form, and central to their application is a knowledge of the diffusion coefficient of a collection of the elements comprising the system. Cohen and Murray [J. Math Biol. 12, 237 (1981)] developed a means of deriving this quantity which has since been largely neglected by model developers, and which is based on a knowledge of the potential energy associated with the mutual interaction between the cells. In this work, we begin by deriving the energy of interaction of biological cells modeled as adhesive, deformable spheres. In so doing, we are able to quantify the equilibrium density of a biological cell aggregate, and also obtain a quantitative estimate of the diffusion coefficient of a collection of cells modeled in this way. In so doing, we are able to use experimental data from single-cell studies of the adhesiveness and cell membrane elasticity of a biological cell to derive the diffusion coefficient of a cell mass composed of a collection of identical cells. This allows us to better inform the parameter values used in reaction-diffusion models of biological systems. We go on to apply this technique to a particular situation: modeling the dynamics of a collection of biological cells which experience strong cell-cell adhesion. In so doing, we derive a nonlinear fourth-order partial differential equation to model this system. We conclude by discussing the practical utility of this work in illuminating the link between the microscopic behavior of individual biological cells and the macroscopic behavior of the aggregate to which they give rise, and also by giving some insights into how the modeling of cell-cell adhesion may be treated mathematically. PMID- 15903698 TI - Anomalous swelling of lipid bilayer stacks is caused by softening of the bending modulus. AB - Arrays of bilayers of the lipid dimyristoylphosphatidylcholine (DMPC) exhibit anomalous swelling as the temperature decreases from T=27 degrees C toward the main phase transition at T(M) =24 degrees C, within the fluid L(alpha) thermodynamic phase. Analysis of diffuse x-ray scattering data from oriented stacks of biological lipid bilayers now makes it possible to obtain the bending modulus K(C) and the bulk compressibility modulus B separately. We report results that show that the measured bending modulus K(C) for DMPC decreases by almost a factor of 2 between T=27 degrees C and the transition temperature at T(M) =24 degrees C, which is the same temperature range where the anomalous swelling occurs. We also report Monte Carlo simulations that show that the anomalous swelling can be fully accounted for by the measured decrease in K(C) with no changes in the van der Waals or hydration forces. PMID- 15903699 TI - Cluster formation by allelomimesis in real-world complex adaptive systems. AB - Animal and human clusters are complex adaptive systems and many organize in cluster sizes s that obey the frequency distribution D (s) proportional to s( tau). The exponent tau describes the relative abundance of the cluster sizes in a given system. Data analyses reveal that real-world clusters exhibit a broad spectrum of tau values, 0.7 (tuna fish schools) > 1, the behavior corresponds to the weak disorder limit and l scales as l approximately L, while for u << 1 , the behavior corresponds to the strong disorder limit with l approximately L(d(opt) ), where d(opt) =1.22+/-0.01 is the optimal path exponent. (b) In the weak disorder regime, there is a length scale xi approximately a(nu), below which strong disorder and critical percolation characterize the current path. PMID- 15903713 TI - Supersymmetry and models for two kinds of interacting particles. AB - We show that Calogero-Sutherland models for interacting particles have a natural supersymmetric extension. For the construction, we use Jacobians that appear in certain superspaces. Some of the resulting Hamiltonians have a direct interpretation as models for two kinds of interacting particles. One model may serve to describe interacting electrons in a lower and upper band of a one dimensional semiconductor, another model corresponds to two kinds of particles confined to two perpendicular spatial directions with an interaction involving tensor forces. PMID- 15903714 TI - Synchronization and desynchronization of self-sustained oscillators by common noise. AB - We consider the effect of external noise on the dynamics of limit cycle oscillators. The Lyapunov exponent becomes negative under influence of small white noise, what means synchronization of two or more identical systems subject to common noise. We analytically study the effect of small non-identities in the oscillators and in the noise, and derive statistical characteristics of deviations from the perfect synchrony. Large white noise can lead to desynchronization of oscillators, provided they are non-isochronous. This is demonstrated for the Van der Pol-Duffing system. PMID- 15903715 TI - Encryption with synchronized time-delayed systems. AB - We propose a new communication scheme that uses time-delayed chaotic systems with delay time modulation. In this method, the transmitter encodes a message as an additional modulation of the delay time and then the receiver decodes the message by tracking the delay time. We demonstrate our communication scheme in a system of coupled logistic maps. Also we discuss the error of the transferred message due to an external noise and present its correction method. PMID- 15903716 TI - Two-fluid model based on the lattice Boltzmann equation. AB - A two-fluid model for dispersed two-phase flows based on the lattice Boltzmann equation (LBE) is proposed. Two sets of LBEs are used to describe the two phases. The continuity and Navier-Stokes equations for the continuum-based two-fluid model are obtained from the LBEs by adding a pressure term. The phase hold-up is readily calculated from the partial pressure of each phase because the two phases are described by the ideal gas equation of state. A simulated laminar gas-liquid two-phase flow gave good agreement with the results calculated by the conventional continuum-based two-fluid model. PMID- 15903717 TI - Ordering of dust particles in dusty plasmas under microgravity. AB - Structure formation of dust particles in dusty plasmas under microgravity has been simulated by the molecular dynamics method. It is shown that, at low temperatures, dust particles are organized into layered spherical shells. The number of shells is a function of the system size and the strength of screening by ambient plasma particles, while the dependency on the latter is much weaker. In the simulation, the condition of the charge neutrality satisfied by the system of dust particles and plasma particles is properly taken into account. PMID- 15903718 TI - Spatial and temporal turbulent velocity and vorticity power spectra from sound scattering. AB - By performing sound-scattering measurements with a detector array consisting of 62 elements in a flow between two counter-rotating disks we obtain the energy and vorticity power spectra directly in both spatial and temporal domains. Fast accumulated statistics and a large signal-to-noise ratio allow us to get high quality data rather effectively and to test scaling laws in details. PMID- 15903719 TI - Fast sampling algorithm for Lie-Trotter products. AB - A fast algorithm for path sampling in path-integral Monte Carlo simulations is proposed. The algorithm utilizes the Levy-Ciesielski implementation of Lie Trotter products to achieve a mathematically proven computational cost of n log2(n) with the number of time slices n, despite the fact that each path variable is updated separately, for reasons of optimality. In this respect, we demonstrate that updating a group of random variables simultaneously results in loss of efficiency. PMID- 15903720 TI - Identifying communities within energy landscapes. AB - Potential energy landscapes can be represented as a network of minima linked by transition states. The community structure of such networks has been obtained for a series of small Lennard-Jones (LJ) clusters. This community structure is compared to the concept of funnels in the potential energy landscape. Two existing algorithms have been used to find community structure, one involving removing edges with high betweenness, the other involving optimization of the modularity. The definition of the modularity has been refined, making it more appropriate for networks such as these where multiple edges and self-connections are not included. The optimization algorithm has also been improved, using Monte Carlo methods with simulated annealing and basin hopping, both often used successfully in other optimization problems. In addition to the small clusters, two examples with known heterogeneous landscapes, the 13-atom cluster (LJ13) with one labeled atom and the 38-atom cluster (LJ38) , were studied with this approach. The network methods found communities that are comparable to those expected from landscape analyses. This is particularly interesting since the network model does not take any barrier heights or energies of minima into account. For comparison, the network associated with a two-dimensional hexagonal lattice is also studied and is found to have high modularity, thus raising some questions about the interpretation of the community structure associated with such partitions. PMID- 15903721 TI - Voting and catalytic processes with inhomogeneities. AB - We consider the dynamics of the voter model and of the monomer-monomer catalytic process in the presence of many "competing" inhomogeneities and show, through exact calculations and numerical simulations, that their presence results in a non-trivial fluctuating steady state whose properties are studied and turn out to specifically depend on the dimensionality of the system, the strength of the inhomogeneities, and their separating distances. In fact, in arbitrary dimensions, we obtain an exact (yet formal) expression of the order parameters (magnetization and concentration of adsorbed particles) in the presence of an arbitrary number n of inhomogeneities ("zealots" in the voter language) and formal similarities with suitable electrostatic systems are pointed out. In the non-trivial cases n = 1,2, we explicitly compute the static and long-time properties of the order parameters and therefore capture the generic features of the systems. When n > 2 , the problems are studied through numerical simulations. In one spatial dimension, we also compute the expressions of the stationary order parameters in the completely disordered case, where n is arbitrary large. Particular attention is paid to the spatial dependence of the stationary order parameters and formal connections with electrostatics. PMID- 15903722 TI - Anomalous rotational relaxation: a fractional Fokker-Planck equation approach. AB - In this study we have analytically obtained the relaxation function in terms of rotational correlation functions based on Brownian motion for complex disordered systems in a stochastic framework. We found out that the rotational relaxation function has a fractional form for complex disordered systems, which indicates that relaxation has nonexponential character and obeys the Kohlrausch-William Watts law, following the Mittag-Leffler decay. PMID- 15903723 TI - Phase behavior of Ising mixtures. AB - We present phase diagrams that were calculated both in mean-field theory and via Monte Carlo (MC) simulations for binary mixtures of a ferromagnetic Ising fluid and a nonmagnetic fluid (Ising mixtures) in the absence of an external field. We look at both the simple ideal Ising mixture, consisting of an ideal Ising fluid and a hard-sphere fluid, as well as at the general case with one component being a nonideal Ising fluid and the other a van der Waals fluid. It is shown that the mean-field phase diagram of the ideal Ising mixture in the limit of infinite pressure is identical to that of the Blume-Capel model for 3He-4He mixtures. The MC phase diagrams were obtained using the Gibbs ensemble, the cumulant intersection technique, and the multi-histogram re-weighting method, adapted to the semi-grand ensemble. The results are qualitatively compared with mean-field theory, and both types of tri-critical lines occurring there are verified in the computer simulations. PMID- 15903724 TI - Local symmetries and order-disorder transitions in small macroscopic Wigner islands. AB - The influence of local order on the disordering scenario of small Wigner islands is discussed. A first disordering step is put in evidence by the time correlation functions and is linked to individual excitations resulting in configuration transitions, which are very sensitive to the local symmetries. This is followed by two other transitions, corresponding to orthoradial and radial diffusion, for which both individual and collective excitations play a significant role. Finally, we show that, contrary to large systems, the focus that is commonly made on collective excitations for such small systems through the Lindemann criterion has to be made carefully in order to clearly identify the relative contributions in the whole disordering process. PMID- 15903725 TI - Work extraction in the spin-boson model. AB - We show that work can be extracted from a two-level system (spin) coupled to a bosonic thermal bath. This is possible due to different initial temperatures of the spin and the bath, both positive (no spin population inversion), and is realized by means of a suitable sequence of sharp pulses applied to the spin. The extracted work can be of the order of the response energy of the bath, therefore much larger than the energy of the spin. Moreover, the efficiency of extraction can be very close to its maximum, given by the Carnot bound, at the same time the overall amount of the extracted work is maximal. Therefore, we get a finite power at efficiency close to the Carnot bound. The effect comes from the back-reaction of the spin on the bath, and it survives for a strongly disordered (inhomogeneously broadened) ensemble of spins. It is connected with generation of coherences during the work-extraction process, and we deduced it in an exactly solvable model. All the necessary general thermodynamical relations are deduced from the first principles of quantum mechanics and connections are made with processes of lasing without inversion and with quantum heat engines. PMID- 15903726 TI - Minimal work principle: proof and counter-examples. AB - The minimal work principle states that work done on a thermally isolated equilibrium system is minimal for adiabatically slow (reversible) realization of a given process. This principle, one of the formulations of the second law, is studied here for finite (possibly large) quantum systems interacting with macroscopic sources of work. It is shown to be valid as long as the adiabatic energy levels do not cross. If level crossing does occur, counter-examples are discussed, showing that the minimal work principle can be violated and that optimal processes are neither adiabatically slow nor reversible. The results are corroborated by an exactly solvable model. PMID- 15903727 TI - Phase transitions of extended-range probabilistic cellular automata with two absorbing states. AB - We study phase transitions in a long-range one-dimensional cellular automaton with two symmetric absorbing states. It includes and extends several other models, like the Ising and Domany-Kinzel ones. It is characterized by competing ferromagnetic linear and antiferromagnetic nonlinear couplings. Despite its simplicity, this model exhibits an extremely rich phase diagram. We present numerical results and mean-field approximations. PMID- 15903728 TI - Blocking temperature in magnetic nanoclusters. AB - A recent study of non-extensive phase transitions in nuclei and nuclear clusters needs a probability model compatible with the appropriate Hamiltonian. For magnetic molecules a representation of the evolution by a Markov process achieves the required probability model that is used to study the probability density function (PDF) of the order parameter, i.e., the magnetization. The existence of one or more modes in this PDF is an indication for the super-paramagnetic transition of the cluster. This allows us to determine the factors that influence the blocking temperature, i.e., the temperature related to the change of the number of modes in the density. It turns out that for our model, rather than the evolution of the system implied by the Hamiltonian, the high temperature density of the magnetization is the important factor for the temperature of the transition. We find that an initial probability density function with a high entropy leads to a magnetic cluster with a high blocking temperature. PMID- 15903729 TI - Mean-field results for the two-component model. AB - In previous papers we have introduced a new dynamical model of Ising spins: the two-component (TC) model. In this paper we formulate a mean-field version of the TC model by putting it on a complete graph. With such an approach we are able to describe the kinetics in terms of a one-dimensional stochastic process with hopping probabilities depending on the magnetization. This allows us to understand the differences in relaxation between phases observed previously in computer simulations for the TC model on the square lattice. PMID- 15903730 TI - Generalized mean-field theory for Ising spins in small world networks. AB - A generalization of mean-field theory for random systems is described. The results of that analytic model could be reconciled with the results of numerical calculations of the Curie temperature for a system of Ising spins in small world (SW) networks by introducing the effective interaction energy associated with long-range links which exceeds the real energy of spin interaction. Such a model describes qualitatively well the increasing Curie temperature T(C) with the growth of the long-range links fraction p in the two-dimensional SW system with fixed coordination number. On the basis of simple physical considerations, concentration dependences T(C)(p) are found for SW systems of different dimensions. PMID- 15903731 TI - Finite-size-scaling analysis of the XY universality class between two and three dimensions: an application of Novotny's transfer-matrix method. AB - Based on Novotny's transfer-matrix method, we simulated the (stacked) triangular Ising anti-ferromagnet embedded in the space with the dimensions variable in the range 2 < or = d < or = 3. Our aim is to investigate the criticality of the XY universality class for 2 < or = d < or = 3. For that purpose, we employed an extended version of the finite-size-scaling analysis developed by Novotny, who utilized this scheme to survey the Ising criticality (ferromagnet) for 1 < or = d < or = 3. Diagonalizing the transfer matrix for the system sizes N up to N = 17 , we calculated the d -dependent correlation-length critical exponent nu(d). Our simulation result nu(d) appears to interpolate smoothly the known two limiting cases, namely, the Kosterlitz-Thouless (KT) and d = 3 XY universality classes, and the intermediate behavior bears close resemblance to that of the analytical formula via the 1/N-expansion technique. Methodological details including the modifications specific to the present model are reported. PMID- 15903732 TI - Observable dependent quasi-equilibrium in slow dynamics. AB - We present examples demonstrating that quasi-equilibrium fluctuation-dissipation behavior at short time differences is not a generic feature of systems with slow nonequilibrium dynamics. We analyze in detail the nonequilibrium fluctuation dissipation ratio X(t, t(w)) associated with a defect-pair observable in the Glauber-Ising spin chain. It turns out that X not = 1 throughout the short-time regime and in particular X(t(w), t(w)) = 3/4 for t(w) --> infinity. The analysis is extended to observables detecting defects at a finite distance from each other, where similar violations of quasi-equilibrium behavior are found. We discuss our results in the context of metastable states, which suggests that a violation of short-time quasi-equilibrium behavior could occur in general glassy systems for appropriately chosen observables. PMID- 15903733 TI - Multiscale complexity of correlated Gaussians. AB - We apply a recently developed measure of multiscale complexity to the Gaussian model consisting of continuous spins with bilinear interactions for a variety of interaction matrix structures. We find two universal behaviors of the complexity profile. For systems with variables that are not frustrated, an exponential decay of multiscale complexity in the disordered regime shows the presence of small scale fluctuations and a logarithmically diverging profile of fixed shape near the critical point describes the spectrum of collective modes. For frustrated variables, oscillations in complexity indicate the presence of global or local constraints. These observations show that the multiscale complexity may be a useful tool for interpreting the underlying structure of systems for which pair correlations can be measured. PMID- 15903734 TI - Randomly accelerated particle in a box: mean absorption time for partially absorbing and inelastic boundaries. AB - Consider a particle which is randomly accelerated by Gaussian white noise on the line segment 0 A + nB with n = 1 or 2. The first level belongs to the directed percolation or the parity-conserving (PC) universality class, the second to PC. If both levels are critical, the active region of the second level becomes heterogeneous. In the so-called coupled region the first level feeds particles to the second, while in the uncoupled region the second level evolves autonomously. Measuring dynamic critical exponents in both regions, we show to what extent the critical behavior of the second level depends on the universality class of the first. These results suggest a simple criterion for the emergence of unusual critical behavior of unidirectionally coupled nonequilibrium systems. PMID- 15903742 TI - Heterogeneous beliefs, segregation, and extremism in the making of public opinions. AB - The connection between contradictory public opinions, heterogeneous beliefs, and the emergence of majority- or minority-induced extremism is studied, extending our former two-state dynamic opinion model. Agents are attached to a social cultural class. At each step they are distributed randomly in different groups within their respective classes to evolve locally by majority rule. In case of a tie the group adopts one or another opinion with respective probabilities k and (1-k) . The value of k accounts for the average of individual biases driven by the existence of heterogeneous beliefs within the corresponding class. It may vary from class to class. The process leads to extremism with a full polarization of each class along one opinion. For homogeneous classes the extremism can be along the initial minority making it minority induced. In contrast, heterogeneous classes exhibit more balanced dynamics, which results in a majority-induced extremism. Segregation among subclasses may produce a coexistence of opinions at the class level, thus averting global extremism. Insight into the existence of contradictory public opinions in similar social-cultural neighborhoods is given. PMID- 15903743 TI - Criticality in the Burridge-Knopoff model. AB - Criticality is a potential origin of the scale invariance observed in the Gutenberg-Richter law for earthquakes. In support of this hypothesis, the Burridge-Knopoff (BK) model of an earthquake fault system is known to exhibit a dynamic phase transition, but the critical nature of the transition is uncertain. Here it is shown that the BK model exhibits a dynamic transition from large-scale stick-slip to small-scale creep motion and through a finite size scaling analysis the critical nature of this transition is established. The order parameter describing the critical transition suggests that the Olami-Feder-Christensen model may be tuned to criticality through its assumptions describing the relaxation of the system. PMID- 15903744 TI - Diffuse-interface model for rapid phase transformations in nonequilibrium systems. AB - A thermodynamic approach to rapid phase transformations within a diffuse interface in a binary system is developed. Assuming an extended set of independent thermodynamic variables formed by the union of the classic set of slow variables and the space of fast variables, we introduce finiteness of the heat and solute diffusive propagation at the finite speed of the interface advancing. To describe transformations within the diffuse interface, we use the phase-field model which allows us to follow steep but smooth changes of phase within the width of the diffuse interface. Governing equations of the phase-field model are derived for the hyperbolic model, a model with memory, and a model of nonlinear evolution of transformation within the diffuse interface. The consistency of the model is proved by the verification of the validity of the condition of positive entropy production and by outcomes of the fluctuation dissipation theorem. A comparison with existing sharp-interface and diffuse interface versions of the model is given. PMID- 15903745 TI - Monte Carlo renormalization: the triangular Ising model as a test case. AB - We test the performance of the Monte Carlo renormalization method in the context of the Ising model on a triangular lattice. We apply a block-spin transformation which allows for an adjustable parameter so that the transformation can be optimized. This optimization purportedly brings the fixed point of the transformation to a location where the corrections to scaling vanish. To this purpose we determine corrections to scaling of the triangular Ising model with nearest- and next-nearest-neighbor interactions by means of transfer-matrix calculations and finite-size scaling. We find that the leading correction to scaling just vanishes for the nearest-neighbor model. However, the fixed point of the commonly used majority-rule block-spin transformation appears to lie well away from the nearest-neighbor critical point. This raises the question whether the majority rule is suitable as a renormalization transformation, because the standard assumptions of real-space renormalization imply that corrections to scaling vanish at the fixed point. We avoid this inconsistency by means of the optimized transformation which shifts the fixed point back to the vicinity of the nearest-neighbor critical Hamiltonian. The results of the optimized transformation in terms of the Ising critical exponents are more accurate than those obtained with the majority rule. PMID- 15903746 TI - Interacting damage models mapped onto Ising and percolation models. AB - We introduce a class of damage models on regular lattices with isotropic interactions between the broken cells of the lattice. Quasi-static fiber bundles are an example. The interactions are assumed to be weak, in the sense that the stress perturbation from a broken cell is much smaller than the mean stress in the system. The system starts intact with a surface-energy threshold required to break any cell sampled from an uncorrelated quenched-disorder distribution. The evolution of this heterogeneous system is ruled by Griffith's principle which states that a cell breaks when the release in potential (elastic) energy in the system exceeds the surface-energy barrier necessary to break the cell. By direct integration over all possible realizations of the quenched disorder, we obtain the probability distribution of each damage configuration at any level of the imposed external deformation. We demonstrate an isomorphism between the distributions so obtained and standard generalized Ising models, in which the coupling constants and effective temperature in the Ising model are functions of the nature of the quenched-disorder distribution and the extent of accumulated damage. In particular, we show that damage models with global load sharing are isomorphic to standard percolation theory and that damage models with a local load sharing rule are isomorphic to the standard Ising model, and draw consequences thereof for the universality class and behavior of the autocorrelation length of the breakdown transitions corresponding to these models. We also treat damage models having more general power-law interactions, and classify the breakdown process as a function of the power-law interaction exponent. Last, we also show that the probability distribution over configurations is a maximum of Shannon's entropy under some specific constraints related to the energetic balance of the fracture process, which firmly relates this type of quenched-disorder based damage model to standard statistical mechanics. PMID- 15903747 TI - Two-parameter deformations of logarithm, exponential, and entropy: a consistent framework for generalized statistical mechanics. AB - A consistent generalization of statistical mechanics is obtained by applying the maximum entropy principle to a trace-form entropy and by requiring that physically motivated mathematical properties are preserved. The emerging differential-functional equation yields a two-parameter class of generalized logarithms, from which entropies and power-law distributions follow: these distributions could be relevant in many anomalous systems. Within the specified range of parameters, these entropies possess positivity, continuity, symmetry, expansibility, decisivity, maximality, concavity, and are Lesche stable. The Boltzmann-Shannon entropy and some one-parameter generalized entropies already known belong to this class. These entropies and their distribution functions are compared, and the corresponding deformed algebras are discussed. PMID- 15903748 TI - Exact kinetics of the sol-gel transition. AB - The formation of a gel in a disperse system wherein binary coagulation alone governs the temporal changes of particle mass spectra is studied under the assumption that the coagulation kernel is proportional to the product of masses of coalescing particles. This model is known to reveal the sol-gel transition, i.e., the formation of one giant cluster with the mass comparable to the total mass of the whole system. This paper reports on the exact solution of this model for a finite total mass of the coagulating system. The evolution equation for the generating functional defining all properties of coagulating systems is solved exactly for this particular kernel. The final output is the exact expression for the single-particle mass spectrum as a function of time. The analysis of the spectrum in the thermodynamic limit shows that after a critical time a giant single particle (the gel) appears. Although the concentration of this giant gel particle is zero in the thermodynamic limit, it actively interacts with smaller particles "eating" them and thus growing in mass. Special attention is given to the transition point, where the gel is appearing. It is demonstrated that the sol gel transition reminds the second-order phase transition. The time dependencies of the gel mass, the number concentration, and the second moment of the particle mass spectrum are found. PMID- 15903749 TI - Minimization of the Renyi entropy production in the space-partitioning process. AB - The spontaneous division of space in Fleming-Viot processes is studied in terms of non-extensive thermodynamics. We analyze a system of n different types of Brownian particles confined in a box. Particles of different types annihilate each other when they come into close contact. Each process of annihilation is accompanied by a simultaneous nucleation of a particle of the same type, so that the number of particles of each component remains constant. The system eventually reaches a stationary state, in which the available space is divided into n separate subregions, each occupied by particles of one type. Within each subregion, the particle density distribution minimizes the Renyi entropy production. We show that the sum of these entropy productions in the stationary state is also minimized, i.e., the resulting boundaries between different components adopt a configuration which minimizes the total entropy production. The evolution of the system leads to decreasing of the total entropy production monotonically in time, irrespective of the initial conditions. In some circumstances, the stationary state is not unique-the entropy production may have several local minima for different configurations. In the case of a rectangular box, the existence and stability of different stationary states are studied as a function of the aspect ratio of the rectangle. PMID- 15903750 TI - Quantifying fluctuations in market liquidity: analysis of the bid-ask spread. AB - Quantifying the statistical features of the bid-ask spread offers the possibility of understanding some aspects of market liquidity. Using quote data for the 116 most frequently traded stocks on the New York Stock Exchange over the two-year period 1994-1995, we analyze the fluctuations of the average bid-ask spread S over a time interval deltat. We find that S is characterized by a distribution that decays as a power law P[S>x] approximately x(-zeta(S) ), with an exponent zeta(S) approximately = 3 for all 116 stocks analyzed. Our analysis of the autocorrelation function of S shows long-range power-law correlations, (S(t)S(t + tau)) approximately tau(-mu(s)), similar to those previously found for the volatility. We next examine the relationship between the bid-ask spread and the volume Q, and find that S approximately ln Q; we find that a similar logarithmic relationship holds between the transaction-level bid-ask spread and the trade size. We then study the relationship between S and other indicators of market liquidity such as the frequency of trades N and the frequency of quote updates U, and find S approximately ln N and S approximately ln U. Lastly, we show that the bid-ask spread and the volatility are also related logarithmically. PMID- 15903751 TI - Simulation study of the phase behavior of a planar Maier-Saupe nematogenic liquid. AB - Using extensive Monte Carlo simulations and a simple approximation in density functional theory, we study the phase behavior of a fluid of nematogenic molecules with centers of mass constrained to lie in a plane but with axes free to rotate in any direction, both with and without an external disorienting field perpendicular to the plane. We find that simulation predicts the existence of an order-disorder phase transition belonging to the Berezinskii-Kosterlitz-Thouless type, along with a low temperature gas-liquid transition. In contrast to the simulation results, density functional theory predicts a first-order orientational phase transition coupled continuously with a first-order gas-liquid transition. The approximate theoretical approach qualitatively reproduces the field dependence of the order-disorder and gas-liquid transitions but is far from quantitative. PMID- 15903752 TI - Statistical mechanical load balancer for the web. AB - The maximum entropy principle from statistical mechanics states that a closed system attains an equilibrium distribution that maximizes its entropy. We first show that for graphs with fixed number of edges one can define a stochastic edge dynamic that can serve as an effective thermalization scheme, and hence, the underlying graphs are expected to attain their maximum-entropy states, which turn out to be Erdos-Renyi (ER) random graphs. We next show that (i) a rate-equation based analysis of node degree distribution does indeed confirm the maximum entropy principle, and (ii) the edge dynamic can be effectively implemented using short random walks on the underlying graphs, leading to a local algorithm for the generation of ER random graphs. The resulting statistical mechanical system can be adapted to provide a distributed and local (i.e., without any centralized monitoring) mechanism for load balancing, which can have a significant impact in increasing the efficiency and utilization of both the Internet (e.g., efficient web mirroring), and large-scale computing infrastructure (e.g., cluster and grid computing). PMID- 15903753 TI - Quantum description of Einstein's Brownian motion. AB - A fully quantum treatment of Einstein's Brownian motion is given, stressing in particular the role played by the two original requirements of translational invariance and connection between dynamics of the Brownian particle and atomic nature of the medium. The former leads to a clearcut relationship with a generator of translation-covariant quantum-dynamical semi-groups recently characterized by Holevo, the latter to a formulation of the fluctuation dissipation theorem in terms of the dynamic structure factor, a two-point correlation function introduced in seminal work by Van Hove, directly related to density fluctuations in the medium and therefore to its atomistic, discrete nature. A microphysical expression for the generally temperature dependent friction coefficient is given in terms of the dynamic structure factor and of the interaction potential describing the single collisions. A comparison with the Caldeira-Leggett model is drawn, especially in view of the requirement of translational invariance, further characterizing general structures of reduced dynamics arising in the presence of symmetry under translations. PMID- 15903754 TI - Power law distribution of wealth in population based on a modified Equiluz Zimmermann model. AB - We propose a money-based model for the power law distribution (PLD) of wealth in an economically interacting population. It is introduced as a modification of the Equiluz and Zimmermann (EZ) model for crowding and information transmission in financial markets. Still, it must be stressed that in the EZ model a PLD without exponential correction is obtained only for a particular parameter, while our pattern will give the exact PLD within a wide range. The PLD exponent depends on the model parameters in a non-trivial way and is exactly calculated in this paper. The numerical results are in excellent agreement with the analytic prediction, and also comparable with empirical data of wealth distribution. PMID- 15903755 TI - Monte Carlo studies of the dynamic behavior of the reduction reaction of NO by CO over a surface of clusters supported on a fractal. AB - A Monte Carlo simulation study is made of the dynamic behavior of the reduction reaction of NO by CO, assuming an experimentally representative mechanism, over a catalytic substrate consisting of clusters of active sites supported on a fractal. Various empirical laws are found for the evolution in time of production over various cyclic graphs or trees that represent the various clusters of catalytic sites on the surface. An analysis is made of the phase diagrams and the production achieved versus concentration of the gas phase at different temperatures. PMID- 15903756 TI - Domain statistics in a finite Ising chain. AB - We present a comprehensive study for the statistical properties of random variables that describe the domain structure of a finite Ising chain with nearest neighbor exchange interactions and free boundary conditions. By use of extensive combinatorics we succeed in obtaining the one-variable probability functions for (i) the number of domain walls, (ii) the number of up domains, and (iii) the number of spins in an up domain. The corresponding averages and variances of these probability distributions are calculated and the limiting case of an infinite chain is considered. Analyzing the averages and the transition time between differing chain states at low temperatures, we also introduce a criterion of the ferromagnetic-like behavior of a finite Ising chain. The results can be used to characterize magnetism in monatomic metal wires and atomic-scale memory devices. PMID- 15903757 TI - Renormalization-group and numerical analysis of a noisy Kuramoto-Sivashinsky equation in 1 + 1 dimensions. AB - The long-wavelength properties of a noisy Kuramoto-Sivashinsky (KS) equation in 1 + 1 dimensions are investigated by use of the dynamic renormalization group (RG) and direct numerical simulations. It is shown that the noisy KS equation is in the same universality class as the Kardar-Parisi-Zhang (KPZ) equation in the sense that they have scale invariant solutions with the same scaling exponents in the long-wavelength limit. The RG analysis reveals that the RG flow for the parameters of the noisy KS equation rapidly approach the KPZ fixed point with increasing strength of the noise. This is supplemented by numerical simulations of the KS equation with a stochastic noise, in which scaling behavior close to the KPZ scaling can be observed even in a moderate system size and time. PMID- 15903758 TI - Small world graphs by iterated local edge formation. AB - We study graphs obtained by successive creation and destruction of edges into small neighborhoods of the vertices. Starting with a circle graph of large diameter we obtain small world graphs with logarithmic diameter, high clustering coefficients, and a fat tail distribution for the degree. Only local edge formation processes are involved and no preferential attachment was used. Furthermore, we found an interesting phase transition with respect to the initial conditions. PMID- 15903759 TI - Nonequilibrium temperatures in steady-state systems with conserved energy. AB - We study a class of nonequilibrium lattice models describing local redistributions of a globally conserved quantity, which is interpreted as an energy. A particular subclass can be solved exactly, allowing us to define a statistical temperature T(th) along the same lines as in the equilibrium micro canonical ensemble. We compute the response function and find that when the fluctuation-dissipation relation is linear, the slope T(FD)(-1) of this relation differs from the inverse temperature T(th)(-1). We argue that T(th) is physically more relevant than T(FD), since in the steady-state regime, it takes equal values in two subsystems of a large isolated system. Finally, a numerical renormalization group procedure suggests that all models within the class behave similarly at a coarse-grained level, leading to a parameter that describes the deviation from equilibrium. Quantitative predictions concerning this parameter are obtained within a mean-field framework. PMID- 15903760 TI - Maximal planar networks with large clustering coefficient and power-law degree distribution. AB - In this article, we propose a simple rule that generates scale-free networks with very large clustering coefficient and very small average distance. These networks are called random Apollonian networks (RANs) as they can be considered as a variation of Apollonian networks. We obtain the analytic results of power-law exponent gamma=3 and clustering coefficient C= (46/3)-36 ln 3/2 approximately 0.74, which agree with the simulation results very well. We prove that the increasing tendency of average distance of RANs is a little slower than the logarithm of the number of nodes in RANs. Since most real-life networks are both scale-free and small-world networks, RANs may perform well in mimicking the reality. The RANs possess hierarchical structure as C(k) approximately k(-1) that are in accord with the observations of many real-life networks. In addition, we prove that RANs are maximal planar networks, which are of particular practicability for layout of printed circuits and so on. The percolation and epidemic spreading process are also studied and the comparisons between RANs and Barabasi-Albert (BA) as well as Newman-Watts (NW) networks are shown. We find that, when the network order N (the total number of nodes) is relatively small (as N approximately 10(4)), the performance of RANs under intentional attack is not sensitive to N , while that of BA networks is much affected by N. And the diseases spread slower in RANs than BA networks in the early stage of the susceptible-infected process, indicating that the large clustering coefficient may slow the spreading velocity, especially in the outbreaks. PMID- 15903761 TI - Experimental study of the fluctuation theorem in a nonequilibrium steady state. AB - The fluctuation theorem (FT) quantifies the probability of second law violations in small systems over short time scales. While this theorem has been experimentally demonstrated for systems that are perturbed from an initial equilibrium state, there are a number of studies suggesting that the theorem applies asymptotically in the long time limit to systems in a nonequilibrium steady state. The asymptotic application of the FT to such nonequilibrium steady states has been referred to in the literature as the steady-state fluctuation theorem (or SSFT). In this paper, we demonstrate experimentally the application of the FT to nonequilibrium steady states, using a colloidal particle localized in a translating optical trap. Furthermore, we show, for this colloidal system, that the FT holds under nonequilibrium steady states for all time, and not just in the long time limit, as in the SSFT. PMID- 15903762 TI - Extended Einstein relations with a complex effective temperature in a one dimensional driven lattice gas. AB - We carry out numerical experiments on a one-dimensional driven lattice gas to elucidate the statistical properties of steady states far from equilibrium. By measuring the bulk density diffusion constant D, the conductivity sigma, and the intensity of density fluctuations, chi, we confirm that the Einstein relation Dchi=sigmaT, which is valid in the linear response regime about equilibrium, does not hold in such steady states. Here, T is the environment temperature and the Boltzmann constant is set to unity. Recalling that the Einstein relation provided the first step in the construction of linear response theory, we attempt to extend it to a generalized form valid in steady states far from equilibrium. In order to obtain new relations among measurable quantities, we define a complex effective temperature theta-iphi from studying the static response of the system to a slowly varying potential in space. Replacing T in the Einstein relation by the real part of the effective temperature Theta , we numerically confirm that the relation Dchi=sigmatheta holds in the nonequilibrium steady states far from equilibrium that we study. In addition to this extended form, we find the relation (L/2pi)cchi=sigmaphi , where c represents the propagation velocity of density fluctuations. PMID- 15903763 TI - Generalized entropy arising from a distribution of q indices. AB - It is by now well known that the Boltzmann-Gibbs (BG) entropy can be usefully generalized using the non-extensive entropies, which have been applied to a wide range of phenomena. However, it seems that even more general entropies could be useful in order to describe other complex physical systems, a task which has already been undertaken in the literature. Following this approach, we introduce here a quite general entropy based on a distribution of q indices thus generalizing S(q). We establish some general mathematical properties for the new entropic functional and explore some examples. We also exhibit a procedure for finding, given any entropic functional, the q-indices distribution that produces it. Finally, on the road to establishing a quite general statistical mechanics, we briefly address possible generalized constraints under which the present entropy could be extremized, in order to produce canonical-ensemble-like stationary-state distributions for Hamiltonian systems. PMID- 15903764 TI - Experimental investigations of chaos-assisted tunneling in a microwave annular billiard. AB - We present detailed investigations of the experimental signatures of chaos assisted tunneling in the two-dimensional annular billiard, as already summarized in Phys. Rev. Lett. 84, 867 (2000). We have performed analog experiments with two dimensional, electromagnetic resonators allowing for a direct simulation of the corresponding quantum system. Spectra from a superconducting cavity with a high frequency resolution are combined with electromagnetic intensity distributions of high spatial resolution experimentally determined using a normal conducting twin cavity. Thereby all eigenmodes were obtained with properly identified quantum numbers. Besides distributions of quasi-doublet splittings, which serve as fundamental observables for the tunneling between whispering gallery types of modes, we also focus on the distributions of resonance widths of the doublets. These directly reflect the role of lifetime of certain modes in the tunneling process. Here, as theoretically expected, the class of so-called beach modes is found to play a particular role in mediating between regular and chaotic states to enhance the tunneling strength. This behavior is found in the spectrum and also in the structure of the wave functions. PMID- 15903765 TI - Distribution of resonance strengths in microwave billiards of mixed and chaotic dynamics. AB - A new measure for statistical properties of the wave function components of quantum systems, the distribution of the product of two partial widths, is introduced. It is tested with data obtained in analog experiments with microwave billiards, where the product of two partial widths equals the resonance strengths in the microwave spectra. The billiards are from the family of the Limacons, one with chaotic and two with mixed classical dynamics. For completely chaotic systems the partial widths generically obey a Porter-Thomas distribution. We show that in this case the distribution of their product equals a K0 distribution. While we find deviations of the experimental strength distribution from the K0 distribution for the billiards with mixed dynamics, the distributions agree perfectly for the chaotic billiard, when taking into account the experimental threshold of detection in the theoretical description. Hence, the strength distribution provides another stringent test for the connection between statistical properties of systems with classical chaotic dynamics and random matrix theory. PMID- 15903766 TI - Fluctuational transitions across different kinds of fractal basin boundaries. AB - We study fluctuational transitions in discrete and continuous dynamical systems that have two coexisting attractors in phase space, separated by a fractal basin boundary which may be either locally disconnected or locally connected. Theoretical and numerical evidence is given to show that, in each case, the transition occurs via a unique accessible point on the boundary, both in discrete systems and in flows. The complicated structure of the escape paths inside the locally disconnected fractal basin boundary is determined by a hierarchy of homoclinic points. The interrelation between the mechanism of transitions and the hierarchy is illustrated by consideration of fluctuational transitions in dynamical systems demonstrating "fractal-fractal" basin boundary metamorphosis at some value of a control parameter. The most probable escape path from an attractor, which can be either regular or chaotic, is found for each type of boundary using both statistical analysis of fluctuational trajectories and the Hamiltonian theory of fluctuations. PMID- 15903767 TI - Zeros in single-channel transmission through double quantum dots. AB - By using a simple model we consider single-channel transmission through a double quantum dot that consists of two single dots coupled by a wire of finite length L . Each of the two single dots is characterized by a few energy levels only, and the wire is assumed to have only one level whose energy depends on the length L . The transmission is described by using S matrix theory and the effective non Hermitian Hamilton operator H(eff) of the system. The decay widths of the eigenstates of H(eff) depend strongly on energy. The model explains the origin of the transmission zeros of the double dot that is considered by us. Mostly, they are caused by (destructive) interferences between neighboring levels and are of first order. When, however, both single dots are identical and their transmission zeros are of first order, those of the double dot are of second order. First order transmission zeros cause phase jumps of the transmission amplitude by pi, while there are no phase jumps related to second-order transmission zeros. In this latter case, a phase jump occurs due to the fact that the width of one of the states vanishes when crossing the energy of the transmission zero. The parameter dependence of the widths of the resonance states is determined by the spectral properties of the two single dots. PMID- 15903768 TI - Electric circuit networks equivalent to chaotic quantum billiards. AB - We consider two electric RLC resonance networks that are equivalent to quantum billiards. In a network of inductors grounded by capacitors, the eigenvalues of the quantum billiard correspond to the squared resonant frequencies. In a network of capacitors grounded by inductors, the eigenvalues of the billiard are given by the inverse of the squared resonant frequencies. In both cases, the local voltages play the role of the wave function of the quantum billiard. However, unlike for quantum billiards, there is a heat power because of the resistance of the inductors. In the equivalent chaotic billiards, we derive a distribution of the heat power which describes well the numerical statistics. PMID- 15903769 TI - Finite-element distributions as an apparatus for detection of the Einstein Brillouin-Keller quantized energy levels. AB - Regularities encountered for some systems with non-eneric spacing distributions are discussed and a corollary which connects these regularities with the Einstein Brillouin-Keller (EBK) quantized energy levels is formulated. Properties of quantum systems are investigated with the help of finite elements probability distributions. Analytic properties of three-point finite-element distributions are investigated. Four-point finite-element distributions for an integrable model are presented. Analytic properties of such distributions for chaotic and integrable models are discussed. PMID- 15903770 TI - Macroscopic evidence of microscopic dynamics in the Fermi-Pasta-Ulam oscillator chain from nonlinear time-series analysis. AB - The problem of detecting specific features of microscopic dynamics in the macroscopic behavior of a many-degrees-of-freedom system is investigated by analyzing the position and momentum time series of a heavy impurity embedded in a chain of nearest-neighbor anharmonic Fermi-Pasta-Ulam oscillators. Results obtained in a previous work [M. Romero-Bastida, Phys. Rev. E 69, 056204 (2004)] suggest that the impurity does not contribute significantly to the dynamics of the chain and can be considered as a probe for the dynamics of the system to which the impurity is coupled. The (r,tau) entropy, which measures the amount of information generated by unit time at different scales tau of time and r of the observable, is numerically computed by methods of nonlinear time-series analysis using the position and momentum signals of the heavy impurity for various values of the energy density epsilon (energy per degree of freedom) of the system and some values of the impurity mass M. Results obtained from these two time series are compared and discussed. PMID- 15903771 TI - Scaling laws for noise-induced super-persistent chaotic transients. AB - A super-persistent chaotic transient is characterized by the following scaling law for its average lifetime: tau approximately exp[C(p- p(c))(-alpha)] , where C>0 and alpha>0 are constants, p > or = p(c) is a bifurcation parameter, and p(c) is its critical value. As p approaches p(c) from above, the exponent in the exponential dependence diverges, leading to an extremely long transient lifetime. Historically the possibility of such transient raised the question of whether asymptotic attractors are relevant to turbulence. Here we investigate the phenomenon of noise-induced super-persistent chaotic transients. In particular, we construct a prototype model based on random maps to illustrate this phenomenon. We then approximate the model by stochastic differential equations and derive the scaling laws for the transient lifetime versus the noise amplitude epsilon for both the subcritical (p< p(c)) and the supercritical (p> p(c)) cases. Our results are the following. In the subcritical case where a chaotic attractor exists in the absence of noise, noise-induced transients can be more persistent in the following sense of double-exponential and algebraic scaling: tau approximately exp[K0 exp(K1epsilon(-gamma))] for small noise amplitude epsilon, where K0 >0 , K1 >0, and gamma>0 are constants. The longevity of the transient lifetime in this case is striking. For the supercritical case where there is already a super-persistent chaotic transient, noise can significantly reduce the transient lifetime. These results add to the understanding of the interplay between random and deterministic chaotic dynamics with surprising physical implications. PMID- 15903772 TI - Multimode lasing in two-dimensional fully chaotic cavity lasers. AB - Multimode lasing in a fully chaotic cavity is investigated numerically by using a nonlinear dynamics model. We report a transition process from single-mode lasing to multimode lasing and reveal interactions among the lasing modes. In particular, both mode-pulling and mode-pushing interactions are shown to decrease the number of effective lasing modes. In addition, coexistence of different types of attractors of the final lasing states is numerically confirmed. PMID- 15903773 TI - Coarsening dynamics of the one-dimensional Cahn-Hilliard model. AB - The dynamics of one-dimensional Cahn-Hilliard model is studied. The stationary and particle-type solutions, the bubbles, are perused as a function of initial conditions, boundary conditions, and system size. We characterize the bubble solutions which are involved in the coarsening dynamics and establish the bifurcation scenarios of the system. A set of ordinary differential equation permits us to describe the coarsening dynamics in very good agreement with numerical simulations. We also compare these dynamics with the bubble dynamics deduced from the classical kink interaction computation where our model seems to be more appropriated. In the case of two bubbles, we deduce analytical expressions for the bubble's position and the bubble's width. Besides, a simple description of the ulterior dynamics is presented. PMID- 15903774 TI - Kolmogorov-Sinai entropy for dilute systems of hard particles in equilibrium. AB - In an equilibrium system, the Kolmogorov-Sinai entropy h(KS) equals the sum of the positive Lyapunov exponents, the exponential rates of divergence of infinitesimal perturbations. Kinetic theory may be used to calculate the Kolmogorov-Sinai entropy for dilute gases of many hard disks or spheres in equilibrium at low number density n. The density expansion of h(KS) is NnuA[ln n + B + O(n)], where nu is the single-particle collision frequency. Previous calculations of A were successful. Calculations of B , however, were unsatisfactory. In this paper, I show how the probability distribution of the stretching factor can be determined from a nonlinear differential equation by an iterative method. From this the Kolmogorov-Sinai entropy follows as the average of the logarithm of the stretching factor per unit time. I calculate approximate values of B and compare these to results from existing simulations. The agreement is good. PMID- 15903775 TI - Stripe-hexagon competition in forced pattern-forming systems with broken up-down symmetry. AB - We investigate the response of two-dimensional pattern-forming systems with a broken up-down symmetry, such as chemical reactions, to spatially resonant forcing and propose related experiments. The nonlinear behavior immediately above threshold is analyzed in terms of amplitude equations suggested for a 1:2 and 1:1 ratio between the wavelength of the spatial periodic forcing and the wavelength of the pattern of the respective system. Both sets of coupled amplitude equations are derived by a perturbative method from the Lengyel-Epstein model describing a chemical reaction showing Turing patterns, which gives us the opportunity to relate the generic response scenarios to a specific pattern-forming system. The nonlinear competition between stripe patterns and distorted hexagons is explored and their range of existence, stability, and coexistence is determined. Whereas without modulations hexagonal patterns are always preferred near onset of pattern formation, single-mode solutions (stripes) are favored close to threshold for modulation amplitudes beyond some critical value. Hence distorted hexagons only occur in a finite range of the control parameter and their interval of existence shrinks to zero with increasing values of the modulation amplitude. Furthermore, depending on the modulation amplitude, the transition between stripes and distorted hexagons is either subcritical or supercritical. PMID- 15903776 TI - Statistically relaxing to generating partitions for observed time-series data. AB - We introduce a relaxation algorithm to estimate approximations to generating partitions for observed dynamical time series. Generating partitions preserve dynamical information of a deterministic map in the symbolic representation. Our method optimizes an essential property of a generating partition: avoiding topological degeneracies. We construct an energy-like functional and use a nonequilibrium stochastic minimization algorithm to search through configuration space for the best assignment of symbols to observed data. As each observed point may be assigned a symbol, the partitions are not constrained to an arbitrary parametrization. We further show how to select particular generating partition solutions which also code low-order unstable periodic orbits in a given way, hence being able to enumerate through a number of potential generating partition solutions. PMID- 15903777 TI - Hierarchical crack pattern as formed by successive domain divisions. I. Temporal and geometrical hierarchy. AB - Crack patterns, as they can be observed in the glaze of ceramics or in desiccated mud layers, are formed by successive fractures and divide the two-dimensional plane into distinct domains. On the basis of experimental observation, we develop a description of the geometrical structure of these hierarchical networks. In particular, we show that the essential feature of such a structure can be represented by a genealogical tree of successive domain divisions. This approach allows for a detailed discussion of the relationship between the formation process and the geometric result. We show that--with some restraints--it is possible to reconstruct the history of the system from the geometry of the final pattern. PMID- 15903778 TI - Hierarchical crack pattern as formed by successive domain divisions. II. From disordered to deterministic behavior. AB - Hierarchical crack patterns, such as those formed in the glaze of ceramics or in desiccated layers of mud or gel, can be understood as a successive division of two-dimensional domains. We present an experimental study of the division of a single rectangular domain in drying starch and show that the dividing fracture essentially depends on the domain size, rescaled by the thickness of the cracking layer e. Utilizing basic assumptions regarding the conditions of crack nucleation, we show that the experimental results can be directly inferred from the equations of linear elasticity. Finally, we discuss the impact of these results on hierarchical crack patterns, and in particular the existence of a transition from disordered cracks at large scales--the first ones--to a deterministic behavior at small scales--the last cracks. PMID- 15903779 TI - Complete phase and amplitude synchronization of broadband chaotic optical fields generated by semiconductor lasers subject to optical injection. AB - A direct experimental observation of phase synchronization, amplitude synchronization, and frequency locking for the high-frequency broadband chaotic optical fields of the transmitter and the receiver is demonstrated in a fully optical system, where its chaotic optical wave form is generated through the high speed nonlinearity of semiconductor lasers subject to optical injection. This experimentally achieved chaotic synchronous scenario is verified as identical chaos synchronization by observing several key characteristics of chaos synchronization in this system. The observation at the frequency detuning, the phase sensitivity, and the effect of mismatch at the injection strength from the master laser and match at the laser output power is in good agreement with the theoretical analysis of this system. PMID- 15903780 TI - Self-affine fractal distributions of the bulk density, elastic moduli, and seismic wave velocities of rock. AB - The scale dependence of the bulk density rho and seismic wave velocities V(p) and V(s) and the possibility of the existence of long-range correlations in such properties in field-scale porous media (FSPM) are investigated. We analyze measured data for rho, V(p), and V(s) for nine FSPM, using the maximum entropy method and the wavelet decomposition technique. The analysis indicates the existence of long-range correlations in the data, characterized by self-affine fractal distributions that follow the statistics of the fractional Brownian motion. Therefore, the elastic moduli of the FSPM should also be fractally distributed and contain long-range correlations; our analysis confirms this. The implications of the results for modeling elastic moduli of porous rock, fracture propagation in FSPM, and the interpretation of seismic wave recordings of FSPM are discussed, and the possible deviations from the classical analysis of such phenomena are pointed out. PMID- 15903781 TI - Gain-scheduled controller for the suppression of convection at high Rayleigh number. AB - Recent studies in the feedback control of Rayleigh-Benard convection indicate that one can sustain the no-motion state at a moderate supercritical Rayleigh number (Ra) using only proportional compensation. However, stabilization occurs at a much higher Rayleigh number using linear-quadratic-Gaussian (LQG) control synthesis. The restriction is that the convection model is linear. In this paper, we show that a comparable degree of stabilization is achievable for a fully nonlinear convection state. The process is demonstrated in two stages using a fully nonlinear, 3D Boussinseq model, compensated by a reduced-order LOQ controller and a gain-schedule table. In the first stage a fully-developed convective state is suppressed through the control action at a moderate supercritical Ra. After the residual convection decays to a sufficiently small amplitude, in the second stage, we increase the Ra by a large step and switch the compensator gains using the gain-schedule table. During this change the control action is in place. Our nonlinear simulation results suggest that the nonlinear system can be stabilized to the limit predicted by the linear analysis. The simulation shows that the large Ra jump induces a large transient temperature in the conductive component, which appears to have very small impact on the stabilization. PMID- 15903782 TI - Effects of initial conditions on a wavelet-decomposed turbulent near-wake. AB - The effect of initial conditions on the turbulent structures of various scales in a near-wake has been investigated for two wake generators, i.e., a circular cylinder and screen of 50% solidity, of the same characteristic dimension. Velocity data were obtained at x/h (x and h are the stream-wise distance downstream and the height of the wake generators, respectively) =20 using 16 X wires, eight aligned in the plane of mean shear and eight in the plane parallel to both the cylinder axis and the flow direction. A wavelet multi-resolution technique was used to decompose the velocity data into a number of wavelet components based on the central frequencies, which correspond to the scales of turbulent structures. It was found that the behaviors of intermediate- as well as large-scale structures depend on the initial conditions. In the circular cylinder wake, the large- and intermediate-scale structures make the largest (and comparable) contribution to Reynolds stresses, but in the screen wake there is a significant difference in the contribution between the two types of structures. The two-dimensionality of the wavelet component is also examined. PMID- 15903783 TI - Numerical solutions of the three-dimensional magnetohydrodynamic alpha model. AB - We present direct numerical simulations and alpha -model simulations of four familiar three-dimensional magnetohydrodynamic (MHD) turbulence effects: selective decay, dynamic alignment, inverse cascade of magnetic helicity, and the helical dynamo effect. The MHD alpha model is shown to capture the long wavelength spectra in all these problems, allowing for a significant reduction of computer time and memory at the same kinetic and magnetic Reynolds numbers. In the helical dynamo, not only does the alpha model correctly reproduce the growth rate of magnetic energy during the kinematic regime, it also captures the nonlinear saturation level and the late generation of a large scale magnetic field by the helical turbulence. PMID- 15903784 TI - Experiments and direct numerical simulations of two-dimensional turbulence. AB - Experiments and direct numerical simulations reveal the coexistence of two cascades in two-dimensional grid turbulence. Several features of this flow such as the energy density and the scalar spectra are found to be consistent with well known theoretical predictions. The energy transfer function displays the expected up-scale energy transfers. The vorticity correlation function is logarithmic and thus consistent with recently proposed models. PMID- 15903785 TI - Effects of temporal density variation and convergent geometry on nonlinear bubble evolution in classical Rayleigh-Taylor instability. AB - Effects of temporal density variation and spherical convergence on the nonlinear bubble evolution of single-mode, classical Rayleigh-Taylor instability are studied using an analytical model based on Layzer's theory [Astrophys. J. 122, 1 (1955)]. When the temporal density variation is included, the bubble amplitude in planar geometry is shown to asymptote to integral(t)U(L)(t')rho(t')dt'/rho(t), where U(L) = square root of (g/(C(g)k)) is the Layzer bubble velocity, rho is the fluid density, and C(g) = 3 and C(g) = 1 for the two- and three-dimensional geometries, respectively. The asymptotic bubble amplitude in a converging spherical shell is predicted to evolve as eta approximately etam(-/r0//(lU(L)sp eta/r0), where r0 is the outer shell radius, eta(t) = integral(t)U(L)sp(t')rho(t') r(0)2(t')dt'/rho(t)r(0)2(t), U(L)(sp) = square root of (-r0(t)r0(t)/l), m(t) = rho(t)r(0)3(t), and l is the mode number. PMID- 15903786 TI - Investigation of the dispersion of heavy-particle pairs and Richardson's law using kinematic simulation. AB - In this paper we investigate the dispersion of heavy-particle pairs (particles with inertia in a gravity field). We vary the particles' inertia when there is no gravity, and the particles' drift at constant inertia. We use kinematic simulation (KS) for which large values of the Reynolds number can be achieved. We investigate the pair diffusivity to discuss Richardson's law, and the locality-in scale hypothesis that underlies that law, for different particle drifts and Stokes numbers. The effect of inertia and gravity on Richardson's law and the autocorrelation in time of the pair's separation are studied for an inertial scale ratio of 1000 and different initial separations delta0 . We extend results from F. Nicolleau and J. C. Vassilicos, Phys. Rev. Lett. 90, 024503 (2003) to three-dimensional (3D) KS and the diffusivity-based analysis of F. Nicolleau and G. Yu, Phys. Fluids 16, 2309 (2004) to particles with inertia and gravity. We find that inertia impedes the locality-in-scale hypothesis whereas gravity improves it. However, the overall effect of gravity and inertia is to decrease the pair's diffusivity in the inertial range. PMID- 15903787 TI - Upscaling and reversibility of Taylor dispersion in heterogeneous porous media. AB - Tracer flow in stratified porous media is dominated by the interaction between convective transport and transverse diffusive mixing. By averaging the tracer concentration in the transverse direction, a one-dimensional non-Fickian dispersion model is derived. The model accounts for the relaxation process that reduces the convective transport to dispersive mixing. This process is (short-) time correlated and partially reversible upon reversal of flow direction. For multiscale velocity fields, the relaxation is a multiscale process. To date only single scale processes have been successfully upscaled. Our procedure extends this to multiscale processes, using scale separation. The model parameters can be calculated a priori based on the velocity profile. For periodic flow reversal, the results are essentially the same. Despite the non-Fickian behavior during a cycle, the net contribution of each cycle to the spreading relaxes to a Fickian process in a similar way as for unidirectional flow. The cycle time averaged dispersion coefficient is a monotonically increasing function of the reversal time. It asymptotically converges towards the effective dispersion coefficient in the absence of flow reversal. PMID- 15903788 TI - Anisotropic diffusion across an external magnetic field and large-scale fluctuations in magnetized plasmas. AB - The problem of random motion of charged particles in an external magnetic field is studied under the assumption that the Langevin sources produce anisotropic diffusion in velocity space and the friction force is dependent on the direction of particle motion. It is shown that in the case under consideration, the kinetic equation describing particle transitions in phase space is reduced to the equation with a Fokker-Planck collision term in the general form (non-isotropic friction coefficient and nonzero off-diagonal elements of the diffusion tensor in the velocity space). The solution of such an equation has been obtained and the explicit form of the transition probability is found. Using the obtained transition probability, the mean-square particle displacements in configuration and velocity space were calculated and compared with the results of numerical simulations, showing good agreement. The obtained results are used to generalize the theory of large-scale fluctuations in plasmas to the case of anisotropic diffusion across an external magnetic field. Such diffusion is expected to be observed in the case of an anisotropic k spectrum of fluctuations generating random particle motion (for example, in the case of drift-wave turbulence). PMID- 15903789 TI - Solitary phase-space holes in pair plasmas. AB - We present theoretical and computer simulation studies of the formation and dynamics of solitary phase-space holes in pair plasmas, which can be applied to both electron-positron plasmas and to plasmas containing positively and negatively charged macro-ions or macro-particulates (charged dust grains). We apply our numerical treatment to the parameters used in a recent series of experiments in a pair ion plasma whose constituents are negatively and positively charged fullerene carbon nanotubes. New experiments should be conducted to confirm our theoretical and numerical predictions. PMID- 15903790 TI - Structure of spherical three-dimensional Coulomb crystals. AB - An analysis of the structural properties of three-dimensional Coulomb clusters confined in a spherical parabolic trap is presented. Based on extensive high accuracy computer simulations the shell configurations and energies for particle numbers in the range 60 < or = N < or = 160 are reported. Further, the intrashell symmetry and the lowest metastable configurations are analyzed for small clusters and a different type of excited state that does not involve a change of shell configuration is identified. PMID- 15903791 TI - Nanosecond electrical explosion of thin aluminum wires in a vacuum: experimental and computational investigations. AB - Experimental and computational investigations of nanosecond electrical explosion of a thin Al wire in vacuum are presented. We have demonstrated that increasing the current rate leads to increased energy deposited before voltage collapse. The experimental evidence for synchronization of the wire expansion and light emission with voltage collapse is presented. Hydrocarbons are indicated in optical spectra and their influence on breakdown physics is discussed. The radial velocity of low-density plasma reaches a value of approximately 100 km/s. The possibility of an over-critical phase transition due to high pressure is discussed. A one-dimensional magnetohydrodynamic (MHD) simulation shows good agreement with experimental data. The MHD simulation demonstrates separation of the exploding wire into a high-density cold core and a low-density hot corona as well as fast rejection of the current from the wire core to the corona during voltage collapse. Important features of the dynamics for the wire core and corona follow from the MHD simulation and are discussed. PMID- 15903792 TI - Coupled dust-lattice modes in complex plasmas. AB - The coupling between transverse and longitudinal dust-lattice modes due to the particle-wake interactions and vertical dust charge gradient is considered. It is shown that the dust-lattice waves can be subjected to a specific instability, the criterion for which has been derived. This instability can explain experimentally observed spontaneous excitation of vibrational modes in a plasma crystal when the pressure is decreased below a critical value. PMID- 15903793 TI - Characteristics and scaling of tungsten-wire-array z -pinch implosion dynamics at 20 MA. AB - We present observations for 20-MA wire-array z pinches of an extended wire ablation period of 57%+/-3% of the stagnation time of the array and non-thin shell implosion trajectories. These experiments were performed with 20-mm-diam wire arrays used for the double- z -pinch inertial confinement fusion experiments [M. E. Cuneo, Phys. Rev. Lett. 88, 215004 (2002)] on the Z accelerator [R. B. Spielman, Phys. Plasmas 5, 2105 (1998)]. This array has the smallest wire-wire gaps typically used at 20 MA (209 microm ). The extended ablation period for this array indicates that two-dimensional (r-z) thin-shell implosion models that implicitly assume wire ablation and wire-to-wire merger into a shell on a rapid time scale compared to wire acceleration are fundamentally incorrect or incomplete for high-wire-number, massive (>2 mg/cm) , single, tungsten wire arrays. In contrast to earlier work where the wire array accelerated from its initial position at approximately 80% of the stagnation time, our results show that very late acceleration is not a universal aspect of wire array implosions. We also varied the ablation period between 46%+/-2% and 71%+/-3% of the stagnation time, for the first time, by scaling the array diameter between 40 mm (at a wire-wire gap of 524 mum ) and 12 mm (at a wire-wire gap of 209 microm ), at a constant stagnation time of 100+/-6 ns . The deviation of the wire-array trajectory from that of a thin shell scales inversely with the ablation rate per unit mass: f(m) proportional[dm(ablate)/dt]/m(array). The convergence ratio of the effective position of the current at peak x-ray power is approximately 3.6+/ 0.6:1 , much less than the > or = 10:1 typically inferred from x-ray pinhole camera measurements of the brightest emitting regions on axis, at peak x-ray power. The trailing mass at the array edge early in the implosion appears to produce wings on the pinch mass profile at stagnation that reduces the rate of compression of the pinch. The observation of precursor pinch formation, trailing mass, and trailing current indicates that all the mass and current do not assemble simultaneously on axis. Precursor and trailing implosions appear to impact the efficiency of the conversion of current (driver energy) to x rays. An instability with the character of an m = 0 sausage grows rapidly on axis at stagnation, during the rise time of pinch power. Just after peak power, a mild m = 1 kink instability of the pinch occurs which is correlated with the higher compression ratio of the pinch after peak power and the decrease of the power pulse. Understanding these three-dimensional, discrete-wire implosion characteristics is critical in order to efficiently scale wire arrays to higher currents and powers for fusion applications. PMID- 15903794 TI - Thomson scattering measurements of heat flux from ion-acoustic waves in laser produced aluminum plasmas. AB - Thomson scattering (TS) measurements are performed at different locations in a laser-produced aluminum plasma. Variations of the separation, wavelength shift, and asymmetric distribution of the two ion-acoustic waves are investigated from their spectral-time-resolved TS images. Detailed information on the space-time evolution of the plasma parameters is obtained. Electron distribution and variation of the heat flux in the plasma are also obtained for a steep temperature gradient. PMID- 15903795 TI - Electron drift velocities in mixtures of helium and xenon and experimental verification of corrections to Blanc's law. AB - Measurements of electron drift velocities were performed in pure Xe and He and in a number of mixtures ranging up to 70% of Xe. The data were obtained by using a pulsed Townsend technique over the density-normalized electric field strength E/N between 1 and 100 Td . Even for pure gases there are no data in the entire range covered here, and these data represent an extension of accurate drift velocities to higher E/N. A selection of well-established cross sections for low energies, which was extended to higher energies, led to a reasonably good agreement of the calculated transport coefficients with the available data. At the same time we have applied the standard (common E/N) Blanc's law and two forms of common mean energy (CME, due to Chiflykian) procedures. Blanc's law fails for most mixtures at low and moderate E/N, while the CME procedure is capable of following the experimental data for the mixtures much more closely, and even predicting the negative differential conductivity region when such effect does not exist for pure gases. Thus the present paper also represents an experimental test of procedures to correct the standard Blanc's law. Finally, we have used the data for two mixtures to obtain results for the third mixture and in all cases this procedure gave excellent results even though only the standard Blanc's law was used in the process. PMID- 15903796 TI - Spectral condensation of turbulence in plasmas and fluids and its role in low-to high phase transitions in toroidal plasma. AB - Transitions from turbulence to order are studied experimentally in thin fluid layers and in magnetically confined toroidal plasma. It is shown that turbulence self-organizes through the mechanism of spectral condensation in both systems. The spectral redistribution of the turbulent energy leads to the reduction in the turbulence level, generation of coherent flow, reduction in the particle diffusion, and increase in the system's energy. The higher-order state in the plasma is sustained via the non-local spectral coupling of the linearly unstable spectral range to the large-scale mean flow. Spectral condensation of turbulence is discussed in terms of its role in the low-to-high confinement transitions in toroidal plasma which show similarity with phase transitions. PMID- 15903797 TI - Phonons in a one-dimensional Yukawa chain: dusty plasma experiment and model. AB - Phonons in a one-dimensional chain of charged microspheres suspended in a plasma were studied in an experiment. The phonons correspond to random particle motion in the chain; no external manipulation was applied to excite the phonons. Two modes were observed, longitudinal and transverse. The velocity fluctuations in the experiment are analyzed using current autocorrelation functions and a phonon spectrum. The phonon energy was found to be unequally partitioned among phonon modes in the dusty plasma experiment. The experimental phonon spectrum was characterized by a dispersion relation that was found to differ from the dispersion relation for externally excited phonons. This difference is attributed to the presence of frictional damping due to gas, which affects the propagation of externally excited phonons differently from phonons that correspond to random particle motion. A model is developed and fit to the experiment to explain the features of the autocorrelation function, phonon spectrum, and the dispersion relation. PMID- 15903798 TI - High-gain harmonic generation free-electron laser with variable wavelength. AB - The external seed of the high-gain harmonic generation (HGHG) free-electron laser (FEL) determines the wavelength of the output radiation. Therefore, the tunability of such a laser depends upon the tunability of the seed. In this paper, we present and discuss an alternative scheme for the tunable HGHG FEL wherein the seed's wavelength is fixed and the variations in the wavelength of radiation are achieved by tuning the accelerator. As an illustration, we apply our proposed scheme to the deep ultraviolet free electron laser (DUV FEL) at Brookhaven National Laboratory demonstrating the ability to attain about a +/-10% variation in the wavelength's tuning range. PMID- 15903799 TI - Bunching instability of rotating relativistic electron layers and coherent synchrotron radiation. AB - We study the stability of a collision-less, relativistic, finite-strength, cylindrical layer of charged particles in free space by solving the linearized Vlasov-Maxwell equations and compute the power of the emitted electromagnetic waves. The layer is rotating in an external magnetic field parallel to the layer. This system is of interest to understanding the high brightness temperature of pulsars which cannot be explained by an incoherent radiation mechanism. Coherent synchrotron radiation has also been observed recently in bunch compressors used in particle accelerators. We consider equilibrium layers with a "thermal" energy spread and therefore a nonzero radial thickness. The particles interact with their retarded electromagnetic self-fields. The effect of the betatron oscillations is retained. A short azimuthal wavelength instability is found which causes a modulation of the charge and current densities. The growth rate is found to be an increasing function of the azimuthal wave number, a decreasing function of the Lorentz factor, and proportional to the square root of the total number of electrons. We argue that the growth of the unstable perturbation saturates when the trapping frequency of electrons in the wave becomes comparable to the growth rate. Owing to this saturation we can predict the radiation spectrum for a given set of parameters. Our predicted brightness temperatures are proportional to the square of the number of particles and scale by the inverse five-third power of the azimuthal wave number which is in rough accord with the observed spectra of radio pulsars. PMID- 15903800 TI - Internal modes in sine-Gordon chain. AB - We address the issue of internal modes of a stable kink in a discrete sine-Gordon equation. The aim of the present study is to elucidate the effects due to the detachment of the frequency dependence of antisymmetric internal mode from the spectrum. We analyze the frequencies of the lowest modes as functions of both the number of sites and the discreteness parameter. Using a simplified approach we explain the origin of the spectrum peculiarity, which arises when the frequency dependence detaches from the quasi-continuous spectrum at some value of the intersite coupling. PMID- 15903801 TI - Reversible soliton motion. AB - We show that spatial solitons on either phase- or amplitude-modulated backgrounds can change their direction of motion according to the modulation frequency. A soliton may, therefore, move up or down phase gradients or remain motionless regardless of where it is in relation to the background modulation. The general theory is in good agreement with numerical results in a variety of nonlinear systems. PMID- 15903802 TI - TE and TM guided modes in an air waveguide with negative-index-material cladding. AB - We numerically demonstrate that a planar waveguide in which the inner layer is a gas with refractive index n0 = 1, sandwiched between two identical semi-infinite layers of a negative index material, can support both transverse electric and transverse magnetic guided modes with low losses. Recent developments in the design of metamaterials with an effective negative index suggest that this waveguide could operate in the infrared region of the spectrum. PMID- 15903803 TI - Postionization regimes of femtosecond laser pulses self-channeling in air. AB - An optical self-guiding of femtosecond filaments in air is identified in a regime where plasma generation ceases to support the self-channeling process. Group velocity dispersion is shown to keep the beam temporally and spatially localized upon a few meters by taking over the ionization of air molecules, once the pulse peak power becomes close to the self-focusing threshold. In this regime, the pulse undergoes slow splitting events that maintain a residual self-guiding with light intensities as high as 10 TW/cm2, as soon as the electron plasma density has fallen down below 10(15) cm(-3). PMID- 15903804 TI - Photonic band gap templating using optical interference lithography. AB - We describe the properties of three families of inversion-symmetric, large photonic band-gap (PBG) template architectures defined by iso-intensity surfaces in four beam laser interference patterns. These templates can be fabricated by optical interference (holographic) lithography in a suitable polymer photo resist. PBG materials can be synthesized from these templates using two stages of infiltration and inversion, first with silica and second with silicon. By considering point and space group symmetries to produce laser interference patterns with the smallest possible irreducible Brillouin zones, we obtain laser beam intensities, directions, and polarizations which generate a diamond-like (fcc) crystal, a novel body-centered cubic (bcc) architecture, and a simple-cubic (sc) structure. We obtain laser beam parameters that maximize the intensity contrasts of the interference patterns. This optimizes the robustness of the holographic lithography to inhomogeneity in the polymer photo-resist. When the optimized iso-intensity surface defines a silicon to air boundary (dielectric contrast of 11.9 to 1), the fcc, bcc, and sc crystals have PBG to center frequency ratios of 25%, 21%, and 11%, respectively. A full PBG forms for the diamond-like crystal when the refractive index contrast exceeds 1.97 to 1. We illustrate a non-inversion symmetric PBG architecture that interpolates between a simple fcc structure and a diamond network structure. This crystal exhibits two distinct and complete photonic band gaps. We also describe a generalized class of tetragonal photonic crystals that interpolate between and extrapolate beyond the diamond-like crystal and the optimized bcc crystal. We demonstrate the extent to which the resulting PBG materials are robust against perturbations to the laser beam amplitudes and polarizations, and template inhomogeneity. The body centered cubic structure exhibits the maximum robustness overall. PMID- 15903805 TI - Effect of eigenmodes on the optical transmission through one-dimensional random media. AB - Using the transfer matrix method as well as numerical solutions to the one dimensional wave equation derived from the Maxwell equations, we study the transition from a photonic crystal with a finite size to a random dielectric medium. We examine the validity of the Kronig-Penney model for a finite size crystal and analyze the spatial structure of the eigenmodes as the crystal is made more irregular. PMID- 15903806 TI - Supersymmetric pairing of kinks for polynomial nonlinearities. AB - We show how one can obtain kink solutions of ordinary differential equations with polynomial nonlinearities by an efficient factorization procedure directly related to the factorization of their nonlinear polynomial part. We focus on reaction-diffusion equations in the traveling frame and damped-anharmonic oscillator equations. We also report an interesting pairing of the kink solutions, a result obtained by reversing the factorization brackets in the supersymmetric quantum-mechanical style. In this way, one gets ordinary differential equations with a different polynomial nonlinearity possessing kink solutions of different width but propagating at the same velocity as the kinks of the original equation. This pairing of kinks could have many applications. We illustrate the mathematical procedure with several important cases, among which are the generalized Fisher equation, the FitzHugh-Nagumo equation, and the polymerization fronts of microtubules. PMID- 15903807 TI - Vector solution of the diffraction task using the Hertz vector. AB - A large class of diffraction problems can be solved on the basis of the Huygens principle. However, methods of solving diffraction problems based on this principle exhibit narrow boundaries of applicability. The goal of the present work is to offer a relatively simple physically based and mathematically strict "dipole wave" vector theory of non-paraxial diffraction of electromagnetic radiation which allows analytical solutions of typical diffraction problems. The suggested theory logically retains the wave approach used in the Kirchhoff method and does not exhibit strict limitations to applicability inherent in the Kirchhoff integral. The diffraction problem is solved by using the Hertz vector in the Kirchhoff integral instead of the field vector. The method efficiency is illustrated in several examples. Analytical solutions of diffraction base problems have been obtained for linearly polarized radiation on an infinite slit and on various-shaped holes at an arbitrary angle of incidence and polarization. It was shown the possibility of vector addition particular solutions to obtain diffraction patterns from several holes. The diffraction of radiation with azimuthal and radial directions of polarization on a ring slit is also considered. The main qualitative feature of the obtained solutions is the presence of "poles" one or two points of zero field in the diffraction pattern which are superimposed on the common system of light and dark fringes. The poles are located along electrical field vector directions. The vector analytical formulas describing the propagation of some laser beams in the free space have been obtained too. The solutions of the diffractive problems satisfy the Maxwell equations and the reciprocity principle. PMID- 15903808 TI - Theory of optical spontaneous emission rates in layered structures. AB - This paper presents expressions for the optical power and fields radiated by an oscillating dipole in layered structures, with loss or gain, that are easy to evaluate. For structures without loss or gain, it is discussed that the radiated power is a direct measure for the local density of modes, whereas for structures with loss or gain, this correspondence no longer holds due to interaction between the dipole and evanescent modes. The presented theory is illustrated with a number of examples. PMID- 15903809 TI - Retrieval of the effective constitutive parameters of bianisotropic metamaterials. AB - We propose a method to retrieve the effective constitutive parameters of a slab of bianisotropic metamaterial composed of split-ring resonators from the measurement of the S parameters. Analytical inversion equations are derived for homogeneous loss-less bianisotropic media, and a numerical retrieval approach is presented for the case of lossy bianisotropic media. The method is verified both analytically and numerically, and it is shown that the results for various split ring resonator metamaterials qualitatively corroborate the conclusions found in published papers. The proposed retrieval method can be used as a valuable tool for the study of anisotropic and bianisotropic properties of metamaterials. PMID- 15903810 TI - Low-loss left-handed materials using metallic magnetic cylinders. AB - We discuss materials based on arrays of metallic magnetic cylindrical structures near ferromagnetic resonance with applied magnetic fields at microwave frequencies. We have found that the materials have a negative refraction index by combining the effective negative permittivity and permeability. Numerical finite difference time-domain simulations were performed, after a very large number of geometries were swept. The simulations reveal that ferromagnetic cylinders, with diameters of 0.1 cm and 0.5 cm apart, and with periodic or random configurations, are left-handed materials with very small losses; i.e., with transmittivity practically unity or no loss. The way to obtain convenient structures and geometries is also discussed. PMID- 15903811 TI - Propagating and evanescent modes in two-dimensional wire media. AB - Electromagnetic waves in an artificial medium formed by two mutually orthogonal lattices of thin ideally conducting straight wires (referred to as a two dimensional wire medium) are considered. An effective medium approach and a full wave method based on the dyadic Green's function and the method of moments are developed. Effects of spatial dispersion, such as the appearance of anisotropy in a square lattice and an additional extraordinary wave, as in crystal optics, are demonstrated. Evanescent waves with complex propagation constants are found. The case when both forward and backward extraordinary waves with respect to an interface exist simultaneously is observed and discussed. The effect of birefringence, so that one extraordinary wave has the wave vector making a positive angle to the interface and the other has the wave vector making a negative angle to the interface, is illustrated. PMID- 15903812 TI - Random-walk statistics in moment-based O(N) tight binding and applications in carbon nanotubes. AB - A computational framework for a moment-based O(N) tight-binding atomistic method is presented, analyzed, and applied to the problem of electronic properties of deformed carbon nanotubes, where N is the number of atoms in the system. The moment-based approach is based on the maximum entropy and kernel polynomial methods for constructing the electronic density of states from local statistical information about the environment around individual atoms. Random-walk statistics are formally presented as the basis for several methods to collect the moments of the density of states in a computationally efficient manner. The computational complexity and accuracy of these methods are systematically analyzed. Using these methods for the problem of deformed carbon nanotubes, it is shown that the computational cost for some cases, per atom, scales as efficiently as O(M log M), where M is the desired number of moments in the expansion of the density of states. These methods are compared to other methods such as direct diagonalization and a Green's function approach. PMID- 15903813 TI - Improved local lattice approach for Coulombic simulations. AB - An improved approach to the simulation of strongly fluctuating Coulomb gases, based on a local lattice technique introduced by Maggs and Rossetto [A.C. Maggs and V. Rossetto, Phys. Rev. Lett. 88, 196402 (2002)], is described and then tested in a problem of biophysical interest. The low acceptance rates for charged particle moves in regimes of physical interest are increased to a serviceable level by use of a coupled particle-field update procedure in the new method. Sensitivity of the results to lattice discretization effects is also studied using asymmetric lattices. PMID- 15903814 TI - Lattice Boltzmann method for homogeneous and heterogeneous cavitation. AB - The Shan and Chen single component multiphase lattice Boltzmann model is used to simulate homogeneous and heterogeneous cavitation. The simulations show excellent agreement with the equation of state for homogeneous cavitation and with energy considerations based on interface formation and bubble expansion for heterogeneous cavitation. PMID- 15903815 TI - Lattice Boltzmann methods for binary mixtures with different molecular weights. AB - Previous authors have suggested lattice Boltzmann methods for binary mixtures. However, these methods are limited to fluids with nearly the same molecular weight. In this work, two modified methods are proposed for simulating fluids with different molecular weights. The first method is based upon the physical principle that particles with different molecular weights move at different lattice speeds (DLS) when at the same temperature. Therefore, different streaming distances are employed for species with different molecular weights. A second method is developed by selecting constants in the equilibrium distribution function in such a way that the speed of sound can be adjusted for each species. In this approach, the species have the same lattice speed (SLS). Using multiscale expansions, the methods are shown to reproduce the appropriate species continuity equation in the macroscopic limit. The accuracy of the methods is evaluated by studying binary diffusion problems. The DLS method is shown to be able to simulate diffusion in fluids with larger ratios of molecular weights relative to the SLS method. PMID- 15903816 TI - First-order interface localization-delocalization transition in thin Ising films using Wang-Landau sampling. AB - Using extensive Monte Carlo simulations, we study the interface localization delocalization transition of a thin Ising film with antisymmetric competing walls for a set of parameters where the transition is strongly first order. This is achieved by estimating the density of states (DOS) of the model by means of Wang Landau sampling (WLS) in the space of energy, using both single-spin-flip as well as N-fold way updates. From the DOS we calculate canonical averages related to the configurational energy, like the internal energy and the specific heat, as well as the free energy and the entropy. By sampling micro-canonical averages during simulations we also compute thermodynamic quantities related to magnetization like the reduced fourth-order cumulant of the order parameter. We estimate the triple temperatures of infinitely large systems for three different film thicknesses via finite size scaling of the positions of the maxima of the specific heat, the minima of the cumulant, and the equal weight criterion for the energy probability distribution. The wetting temperature of the semi-infinite system is computed with help of the Young equation. In the limit of large film thicknesses the triple temperatures are seen to converge toward the wetting temperature of the corresponding semi-infinite Ising model in accordance with standard capillary wave theory. We discuss the slowing down of WLS in energy space as observed for the larger film thicknesses and lateral linear dimensions. In the case of WLS in the space of total magnetization we find evidence that the slowing down is reduced and can be attributed to persisting free energy barriers due to shape transitions. PMID- 15903817 TI - Rarefaction and compressibility effects of the lattice-Boltzmann-equation method in a gas microchannel. AB - A wall equilibrium boundary condition for an implicit lattice-Boltzmann-equation method is proposed to simulate gas flows in a microchannel with rough surface on the characteristic length of gas molecules. The boundary condition is based on the assumption that impinging molecules reach equilibrium with the surface. The molecular mean free path used to define the Knudsen number is determined by the lattice speed and the relaxation time of the lattice-Boltzmann equation. With the wall equilibrium boundary condition and the appropriate relation defined for the Knudsen number and the relaxation time, the computed slip velocity and nonlinear pressure distribution along the microchannel are in excellent agreement with analytical solutions. PMID- 15903818 TI - Levy-Ciesielski random series as a useful platform for Monte Carlo path integral sampling. AB - We demonstrate that the Levy-Ciesielski implementation of Lie-Trotter products enjoys several properties that make it extremely suitable for path-integral Monte Carlo simulations: fast computation of paths, fast Monte Carlo sampling, and the ability to use different numbers of time slices for the different degrees of freedom, commensurate with the quantum effects. It is demonstrated that a Monte Carlo simulation for which particles or small groups of variables are updated in a sequential fashion has a statistical efficiency that is always comparable to or better than that of an all-particle or all-variable update sampler. The sequential sampler results in significant computational savings if updating a variable costs only a fraction of the cost for updating all variables simultaneously or if the variables are independent. In the Levy-Ciesielski representation, the path variables are grouped in a small number of layers, with the variables from the same layer being statistically independent. The superior performance of the fast sampling algorithm is shown to be a consequence of these observations. Both mathematical arguments and numerical simulations are employed in order to quantify the computational advantages of the sequential sampler, the Levy-Ciesielski implementation of path integrals, and the fast sampling algorithm. PMID- 15903819 TI - Molecular dynamics sampling scheme realizing multiple distributions. AB - We present a molecular-dynamics sampling scheme in which any summation of multiple arbitrarily given distributions can be realized deterministically by a certain theoretical guide. Our scheme thus provides a static view of the composition of the distributions, rather than a dynamic view in which some parameter is switched at a certain time in the simulation process. The proposed method induces the orbit to jump automatically among different phase space regions, without the use of any artificial timing parameter. In addition, the proposed method does not require an assumption that the density of states is smooth. We applied it to multiple Tsallis distributions and established a suitable series of parameter values for which the sum of the distributions allows broad sampling. Numerical simulations applied to fundamental models with multi Tsallis distributions showed efficient sampling, characterized by an energy trajectory that was totally different from that associated with each single distribution. PMID- 15903820 TI - Optimization of network robustness to waves of targeted and random attacks. AB - We study the robustness of complex networks to multiple waves of simultaneous (i) targeted attacks in which the highest degree nodes are removed and (ii) random attacks (or failures) in which fractions p(t) and p(r) , respectively, of the nodes are removed until the network collapses. We find that the network design which optimizes network robustness has a bimodal degree distribution, with a fraction r of the nodes having degree k2 = ((k)-1+r)/r and the remainder of the nodes having degree k1=1, where k is the average degree of all the nodes. We find that the optimal value of r is of the order of p(t)/p(r) for p(t)/p(r) << 1. PMID- 15903821 TI - Reciprocity relations between ordinary temperature and the Frieden-Soffer Fisher temperature. AB - Frieden and Soffer conjectured some years ago [Phys. Rev. E 52, 2274 (1995)] the existence of a "Fisher temperature" T(F) that would play, with regards to Fisher's information measure I , the same role that the ordinary temperature T plays in relation to Shannon's logarithmic measure. Here we exhibit the existence of reciprocity relations between T(F) and T and provide an interpretation with reference to the meaning of T(F) for the canonical ensemble. PMID- 15903822 TI - Chaos supported stochastic resonance in a metal-ferroelectric-semiconductor heterostructure. AB - An experimental study is presented on a complex nonlinear system showing a particular type of dynamics that can be interpreted as stochastic resonance. The system consists of a metal-ferroelectric-semiconductor structure, which plays the role of a nonlinear element in an electric circuit with linear resistance, inductance, and capacitance connected in series (RLC series circuit) driven externally by a high-amplitude harmonic voltage source. The system presents various kinds of nonlinear behavior, of which the simplest, consisting of a period-doubling evolution to chaos, is of interest to this study. The broadband intrinsic chaos emerging after a period-doubling sequence exists for a large range of frequencies of the driving voltage. The appearance of the chaotic dynamics is associated with the promotion of a low-frequency harmonic spectral component. This is interpreted as stochastic resonance with intrinsic chaos replacing noise, the usual variable in regular SR. PMID- 15903823 TI - Colored noise in quantum chaos. AB - We derive a set of spectral statistics whose power spectrum is characterized, in the case of chaotic quantum systems, by colored noise 1/f(gamma), where the integer parameter gamma critically depends on the specific energy-level statistic considered. In the case of regular quantum systems these spectral statistics show 1/f(gamma+1) noise. PMID- 15903824 TI - Using geometric control and chaotic synchronization to estimate an unknown model parameter. AB - We present a new parameter estimation procedure for nonlinear systems. Such technique is based on the synchronization between the model and the system whose unknown parameter is wanted. Synchronization is accomplished by controlling the model to make it follow the system. We use geometric nonlinear control techniques to design the control system. These techniques allow us to derive sufficient conditions for synchronization and hence for proper parameter estimation. As an example, this procedure is used to estimate a parameter of an example serving as a model. PMID- 15903825 TI - Dynamic behavior of a liquid metal interface under the influence of a high frequency magnetic field. AB - We present experimental results on the stability of a free liquid metal surface influenced by an alternating magnetic field. The field is generated by an inductor fed by a high-frequency electrical current. The experimental setup consists of an annulus filled with the liquid metal galinstan. We observe three kinds of instabilities: exciting of capillary waves, a static high-amplitude surface deformation, and eventually an electromagnetic pinch at a critical inductor current I(cp). The data show that I(cp) correlates directly with the current frequency and the geometric parameters. PMID- 15903826 TI - Shock-wave formation in Rosenau's extended hydrodynamics. AB - We study the extended hydrodynamics proposed by Rosenau [Phys. Rev. A 40, 7193 (1989)] in the context of a regularization of the Chapman-Enskog expansion. We are able to prove that shock waves appear in finite time in Rosenau's extended Burgers equation, and we discuss the physical implications of this fact and its connection with a possible extension of hydrodynamics to the short-wavelength domain. PMID- 15903827 TI - Simple acoustic multiplexer. AB - Simple structures enabling the multiplexing of acoustic waves are presented. Such structures are constructed out of two monomode acoustic wires and two masses bound together, and to the wires by springs. We show analytically that these simple structures can transfer with selectivity and in one direction one acoustic wavelength from one wire to the other, leaving neighbor acoustic wavelengths unaffected. We give closed-form relations enabling to obtain the values of the relevant physical parameters for this multiplexing phenomena to happen at a chosen wavelength. Finally, we illustrate this general theory by an application. PMID- 15903828 TI - Imaginary-chemical-potential quantum Monte Carlo method for Hubbard molecules. AB - We generalize the imaginary-chemical-potential quantum Monte Carlo (QMC) method proposed by Dagotto [Phys. Rev. B 41, R811 (1990)] to systems without particle hole symmetry. The generalized method is tested by comparing the results of the QMC simulations and exact diagonalization on small Hubbard molecules, such as tetrahedron and truncated tetrahedron. Results of the application of the method to the C60 Hubbard molecule are discussed. PMID- 15903829 TI - Lattice Boltzmann simulation of rarefied gas flows in microchannels. AB - For gas flows in microchannels, slip motion at the solid surface can occur even if the Mach number is negligibly small. Since the Knudsen number of the gas flow in a long microchannel can vary widely and the Navier-Stokes equations are not valid for Knudsen numbers beyond 0.1, an alternative method that can be applicable to continuum, slip and transition flow regimes is highly desirable. The lattice Boltzmann equation (LBE) approach has recently been expected to have such potential. However, some hurdles need to be overcome before it can be applied to simulate rarefied gas flows. The first major hurdle is to accurately model the gas molecule and wall surface interactions. In addition, the Knudsen number needs to be clearly defined in terms of LBE properties to ensure that the LBE simulation results can be checked against experimental measurements and other simulation results. In this paper, the Maxwellian scattering kernel is adopted to address the gas molecule and surface interactions with an accommodation coefficient (in addition to the Knudsen number) controlling the amount of slip motion. The Knudsen number is derived consistently with the macroscopic property based definition. The simulation results of the present LBE model are in quantitative agreement with the established theory in the slip flow regime. In the transition flow regime, the model captures the Knudsen minimum phenomenon qualitatively. Therefore, the LBE can be a competitive method for simulation of rarefied gas flows in microdevices. PMID- 15903830 TI - Global minimum for Thomson's problem of charges on a sphere. AB - Using numerical arguments, we find that for N=306 a tetrahedral configuration (T(h)) and for N=542 a dihedral configuration (D5) are likely the global energy minimum for Thomson's problem of minimizing the energy of N unit charges on the surface of a unit conducting sphere. These would be the largest N by far, outside of the icosadeltahedral series, for which a global minimum for Thomson's problem is known. We also note that the current theoretical understanding of Thomson's problem does not rule out a symmetric configuration as the global minima for N=306 and 542. We explicitly find that analogues of the tetrahedral and dihedral configurations for N larger than 306 and 542, respectively, are not global minima, thus helping to confirm the theory of Dodgson and Moore [Phys. Rev. B 55, 3816 (1997)] that as N grows, dislocation defects can lower the lattice strain of symmetric configurations and concomitantly the energy. As well, making explicit previous work by ourselves and others, for N<1000 we give a full accounting of icosadeltahedral configurations which are not global minima and those which appear to be, and discuss how this listing and our results for the tetahedral and dihedral configurations may be used to refine theoretical understanding of Thomson's problem. PMID- 15903831 TI - Probing pair-correlated fermionic atoms through correlations in atom shot noise. AB - Pair-correlated fermionic atoms are created through dissociation of weakly bound molecules near a magnetic-field Feshbach resonance. We show that correlations between atoms in different spin states can be detected using the atom shot noise in absorption images. Furthermore, using time-of-flight imaging we have observed atom pair correlations in momentum space. PMID- 15903832 TI - Breached superfluidity via p-wave coupling. AB - Anisotropic pairing between fermion species with different Fermi momenta opens two-dimensional areas of gapless excitations, thus producing a spatially homogeneous state with coexisting superfluid and normal fluids. This breached pairing state is stable and robust for arbitrarily small mismatch and weak p-wave coupling. PMID- 15903833 TI - Possibility of a first-order superfluid-Mott-insulator transition of spinor bosons in an optical lattice. AB - We study the superfluid-Mott-insulator transition of antiferromagnetic spin-1 bosons in an optical lattice described by a Bose-Hubbard model. Our variational study with the Gutzwiller variational wave function determines that the superfluid-Mott-insulator transition is a first-order one at a part of the phase boundary curve, contrary to the spinless case. PMID- 15903834 TI - Surface plasmon modes and the Casimir energy. AB - We show the influence of surface plasmons on the Casimir effect between two plane parallel metallic mirrors at arbitrary distances. Using the plasma model to describe the optical response of the metal, we express the Casimir energy as a sum of contributions associated with evanescent surface plasmon modes and propagative cavity modes. In contrast to naive expectations, the plasmonic mode contribution is essential at all distances in order to ensure the correct result for the Casimir energy. One of the two plasmonic modes gives rise to a repulsive contribution, balancing out the attractive contributions from propagating cavity modes, while both contributions taken separately are much larger than the actual value of the Casimir energy. This also suggests possibilities to tailor the sign of the Casimir force via surface plasmons. PMID- 15903835 TI - Energy-time entanglement preservation in plasmon-assisted light transmission. AB - We report on experimental evidence of the preservation of the energy-time entanglement of a pair of photons after a photon-plasmon-photon conversion. This preservation is observed in two different plasmon conversion experiments, namely, extraordinary optical transmission through subwavelength metallic hole arrays and long range surface plasmon propagation in metallic waveguides. Plasmons are shown to coherently exist at two different times separated by much more than their lifetimes. This kind of entanglement involving light and matter is expected to be useful for future processing and storing of quantum information. PMID- 15903836 TI - Robust one-step catalytic machine for high fidelity anticloning and W-state generation in a multiqubit system. AB - We propose a physically realizable machine which can either generate multiparticle W-like states, or implement high-fidelity 1-->M (M=1,2,...infinity) anticloning of an arbitrary qubit state, in a single step. This universal machine acts as a catalyst in that it is unchanged after either procedure, effectively resetting itself for its next operation. It possesses an inherent immunity to decoherence. Most importantly in terms of practical multiparty quantum communication, the machine's robustness in the presence of decoherence actually increases as the number of qubits M increases. PMID- 15903837 TI - Observation of flux reversal in a symmetric optical thermal ratchet. AB - We demonstrate that a cycle of three holographic optical trapping patterns can implement a thermal ratchet for diffusing colloidal spheres and that the ratchet driven transport displays flux reversal as a function of the cycle frequency and the intertrap separation. Unlike previously described ratchet models, the approach we describe involves three equivalent states, each of which is locally and globally spatially symmetric, with spatiotemporal symmetry being broken by the sequence of states. PMID- 15903838 TI - Bounded step superdiffusion in an oriented hexagonal phase. AB - Fluorescence recovery after pattern photobleaching is used to measure the self diffusion of surfactant molecules, along cylinders and perpendicular to their main axis in an oriented hexagonal lyotropic phase. Unexpectedly, while the motion along cylinders is diffusive, a superdiffusive behavior is observed in the direction perpendicular to the cylinder axis. Moreover, varying the lattice parameter, we found that the perpendicular diffusion time is governed only by the number of cylinders to cross, providing experimental evidence for superdiffusion with a bounded step length. PMID- 15903839 TI - Ratchet effect for cold atoms in an optical lattice. AB - The possibility of realizing a directed current for a quantum particle in a flashing asymmetric potential is investigated. It is found that quantum resonances, where the value of the effective Planck constant is equal to an integer or half-integer multiple of pi, give rise to a directed current. The effect should be readily observable in experiments. PMID- 15903840 TI - String picture for a model of frustrated quantum magnets and dimers. AB - We study the effect of quantum dynamics on geometrically frustrated magnets for a transverse field Ising model at finite temperatures. We develop a microscopic derivation of the Landau-Ginzburg-Wilson action for this model and show that it can be interpreted as the free energy of a 3D elastic lattice of noncrossing strings. As a first application, we quantitatively predict the phase diagram and correlations, confirming excellently a key prediction of recent simulations about the existence of unusual phase transitions and an ordered phase. We discuss the implications of our string picture for the understanding of the effect of quenched disorder in such quantum frustrated systems. PMID- 15903841 TI - Ultimate energy density of observable cold baryonic matter. AB - We demonstrate that the largest measured mass of a neutron star establishes an upper bound to the energy density of observable cold baryonic matter. An equation of state-independent expression satisfied by both normal neutron stars and self bound quark matter stars is derived for the largest energy density of matter inside stars as a function of their masses. The largest observed mass sets the lowest upper limit to the density. Implications from existing and future neutron star mass measurements are discussed. PMID- 15903842 TI - Energy spectrum of particles accelerated in relativistic collisionless shocks. AB - We analytically study diffusive particle acceleration in relativistic, collisionless shocks. We find a simple relation between the spectral index s and the anisotropy of the momentum distribution along the shock front. Based on this relation, we obtain s=(3beta(u)-2beta(u)beta(2)(d)+beta(3)(d))/(beta(u)-beta(d)) for isotropic diffusion, where beta(u) (beta(d)) is the upstream (downstream) fluid velocity normalized to the speed of light. This result is in agreement with previous numerical determinations of s for all (beta(u),beta(d)), and yields s=38/9 in the ultrarelativistic limit. The spectrum-anisotropy connection is useful for testing numerical studies and constraining anisotropic diffusion results. It suggests that the spectrum is highly sensitive to the form of the diffusion function for particles traveling along the shock front. PMID- 15903843 TI - Feynman propagator for spin foam quantum gravity. AB - We link the notion causality with the orientation of the spin foam 2-complex. We show that all current spin foam models are orientation independent. Using the technology of evolution kernels for quantum fields on Lie groups, we construct a generalized version of spin foam models, introducing an extra proper time variable. We prove that different ranges of integration for this variable lead to different classes of spin foam models: the usual ones, interpreted as the quantum gravity analogue of the Hadamard function of quantum field theory (QFT) or as inner products between quantum gravity states; and a new class of causal models, the quantum gravity analogue of the Feynman propagator in QFT, nontrivial function of the orientation data, and implying a notion of "timeless ordering". PMID- 15903844 TI - Absence of preclassical solutions in Bianchi I loop quantum cosmology. AB - Loop quantum cosmology, the symmetry reduction of quantum geometry for the study of various cosmological situations, leads to a difference equation for its quantum evolution equation. To ensure that solutions of this equation act in the expected classical manner far from singularities, additional restrictions are imposed on the solution. In this Letter, we consider the Bianchi I model, both the vacuum case and the addition of a cosmological constant, and show using generating function techniques that only the zero solution satisfies these constraints. This implies either that there are technical difficulties with the current method of quantizing the evolution equation, or else loop quantum gravity imposes strong restrictions on the physically allowed solutions. PMID- 15903845 TI - Viscosity in strongly interacting quantum field theories from black hole physics. AB - The ratio of shear viscosity to volume density of entropy can be used to characterize how close a given fluid is to being perfect. Using string theory methods, we show that this ratio is equal to a universal value of variant Planck's over 2pi/4pik(B) for a large class of strongly interacting quantum field theories whose dual description involves black holes in anti-de Sitter space. We provide evidence that this value may serve as a lower bound for a wide class of systems, thus suggesting that black hole horizons are dual to the most ideal fluids. PMID- 15903846 TI - Late time neutrino masses, the LSND experiment, and the cosmic microwave background. AB - Models with low-scale breaking of global symmetries in the neutrino sector provide an alternative to the seesaw mechanism for understanding why neutrinos are light. Such models can easily incorporate light sterile neutrinos required by the Liquid Scintillator Neutrino Detector experiment. Furthermore, the constraints on the sterile neutrino properties from nucleosynthesis and large scale structure can be removed due to the nonconventional cosmological evolution of neutrino masses and densities. We present explicit, fully realistic supersymmetric models, and discuss the characteristic signatures predicted in the angular distributions of the cosmic microwave background. PMID- 15903848 TI - Open charm and beauty at ultrarelativistic heavy ion colliders. AB - Important goals of BNL RHIC and CERN LHC experiments with ion beams include the creation and study of new forms of matter, such as the quark gluon plasma. Heavy quark production and attenuation provide unique tomographic probes of that matter. We predict the suppression pattern of open charm and beauty in Au+Au collisions at RHIC and LHC energies based on the DGLV formalism of radiative energy loss. A cancellation between effects due to the sqrt[s] energy dependence of the high p(T) slope and heavy quark energy loss is predicted to lead to surprising similarity of heavy quark suppression at RHIC and LHC. PMID- 15903849 TI - Half-life of the doubly magic r-process nucleus 78Ni. AB - Nuclei with magic numbers serve as important benchmarks in nuclear theory. In addition, neutron-rich nuclei play an important role in the astrophysical rapid neutron-capture process (r process). 78Ni is the only doubly magic nucleus that is also an important waiting point in the r process, and serves as a major bottleneck in the synthesis of heavier elements. The half-life of 78Ni has been experimentally deduced for the first time at the Coupled Cyclotron Facility of the National Superconducting Cyclotron Laboratory at Michigan State University, and was found to be 110(+100)(-60) ms. In the same experiment, a first half-life was deduced for 77Ni of 128(+27)(-33) ms, and more precise half-lives were deduced for 75Ni and 76Ni of 344(+20)(-24) ms and 238(+15)(-18) ms, respectively. PMID- 15903847 TI - Observation of B+-->K1(1270)+gamma. AB - We report the observation of the radiative decay B+-->K1(1270)(+) gamma using a data sample of 140 fb(-1) taken at the Upsilon(4S) resonance with the Belle detector at the KEKB e+e- collider. We find the branching fraction to be B(B+- >K1(1270)(+)gamma)=(4.3+/-0.9(stat.)+/-0.9(syst.))x10(-5) with a significance of 7.3sigma. We find no significant signal for B+-->K1(1400)(+)gamma and set an upper limit B(B+-->K1(1400)(+)gamma)<1.5 x 10(-5) at the 90% confidence level. We also measure inclusive branching fractions for B+-->K+pi+pi-gamma and B0- >K0pi+pi-gamma in the mass range 1 GeV/c(2) f(m). PMID- 15903889 TI - Ginzburg number of a homopolymer-diblock copolymer mixture covering the 3D-Ising, isotropic Lifshitz, and Brasovskii classes of critical universality. AB - The Ginzburg number Gi of deuterated poly(butadiene) (dPB) and poly(styrene) (PS) homopolymer blend of critical composition mixed with a dPB-PS symmetric diblock copolymer was determined from small angle neutron scattering. A 3 orders of magnitude change of Gi was determined between binary polymer blend and diblock copolymer melt. The strongest change of Gi is observed within the isotropic Lifshitz regime of critical universality occurring over a 3% range of diblock concentration and interpolates the corresponding Gi of the 3D-Ising and Brasovskii regimes. A Lifshitz critical point was not observed consistent with the proposed lower critical dimension d(LCP)=4. PMID- 15903890 TI - Unexpected shish-kebab structure in a sheared polyethylene melt. AB - Scanning electron micrographs of a solvent-extracted sheared polyethylene (PE) blend revealed, for the first time, an unexpected shish-kebab structure with multiple shish. The blend contained 2 wt % of crystallizing ultrahigh molecular weight polyethylene (UHMWPE) and 98 wt % of noncrystallizing PE matrix. The formation of multiple shish was attributed to the coil-stretch transition occurring in sections of UHMWPE chains. Synchrotron x-ray data provided clear evidence of the hypothesis that multiple shish originate from stretched chain sections and kebabs originate from coiled chain sections, following a diffusion controlled crystallization process. PMID- 15903891 TI - Theory for shock dynamics in particle-laden thin films. AB - We present a theory to explain the emergence of a particle-rich ridge observed experimentally in a thin film particle-laden flow on an incline. We derive a lubrication theory for this system which is qualitatively compared to preliminary experimental data. The ridge formation arises from the creation of two shocks due to the differential transport rates of fluid and particles. This parallels recent findings of double shocks in thermal-gravity-driven flow [A. L. Bertozzi, Phys. Rev. Lett. 81, 5169 (1998); J. Sur, Phys. Rev. Lett. 90, 126105 (2003); A. Munch, Phys. Rev. Lett. 91, 016105 (2003)]. However, here the emergence of the shocks arises from a new mechanism involving the settling rates of the species. PMID- 15903892 TI - Mechanics of binding of a single integration-host-factor protein to DNA. AB - We report on a single-molecule experiment where we directly observe local bending of a 76 base pair DNA oligomer caused by specific binding of a single integration host-factor (IHF) protein. The conformational change of the DNA is detected by optically monitoring the displacement of a micron size bead tethered to a surface by the DNA. Since in the bound state the DNA loops around the IHF, a mechanical tension on the DNA tends to eject the protein. We measure how the rate for the protein to fall off the DNA depends on the mechanical tension in the DNA, gaining insight into the energy landscape for this molecular bond. Our method further demonstrates a new paradigm of molecular detection, where ligand binding is detected through the conformational change induced in the probe molecule. Here this allows the detection of single, unlabeled proteins. PMID- 15903893 TI - Control of nanoparticles with arbitrary two-dimensional force fields. AB - An anti-Brownian electrophoretic trap is used to create arbitrary two-dimensional force fields for individual nanoscale objects in solution. The trap couples fluorescence microscopy with digital particle tracking and real-time feedback to generate a position-dependent electrophoretic force on a single nanoparticle. The force may vary over nanometer distances and millisecond times and need not be the gradient of a potential. As illustrations of this technique, I study Brownian motion in harmonic, power-law, and double-well potentials. PMID- 15903894 TI - Prewetting and layering in athermal polymer solutions. AB - Coexistence conditions for prewetting and layering at a hard surface in additive hard sphere polymer solutions, where the solvent particles are smaller than the monomers, have been calculated by density functional methods. Various chain lengths and pressures have been investigated. An unexpected finding is that prewetting in these systems may proceed below the bulk critical pressure. We rationalize this behavior in terms of local properties of the pressure tensor. For longer chains, a different behavior is observed where the systems display a lower wetting pressure, i.e., a low pressure bound for surface wetting. PMID- 15903895 TI - Comment on "Dynamics in a multicomponent plasma near the low-frequency cutoff". PMID- 15903897 TI - Solitary wave complexes in two-component condensates. AB - Axisymmetric three-dimensional solitary waves in uniform two-component mixture Bose-Einstein condensates are obtained as solutions of the coupled Gross Pitaevskii equations with equal intracomponent but varying intercomponent interaction strengths. Several families of solitary wave complexes are found: (1) vortex rings of various radii in each of the components; (2) a vortex ring in one component coupled to a rarefaction solitary wave of the other component; (3) two coupled rarefaction waves; (4) either a vortex ring or a rarefaction pulse coupled to a localized disturbance of a very low momentum. The continuous families of such waves are shown in the momentum-energy plane for various values of the interaction strengths and the relative differences between the chemical potentials of two components. Solitary wave formation, their stability, and solitary wave complexes in two dimensions are discussed. PMID- 15903898 TI - Production efficiency of ultracold feshbach molecules in bosonic and fermionic systems. AB - We investigate the production efficiency of ultracold molecules in bosonic 85Rb and fermionic 40K when the magnetic field is swept across a Feshbach resonance. For adiabatic sweeps of the magnetic field, our novel model shows that the conversion efficiency of both species is solely determined by the phase space density of the atomic cloud, in contrast with a number of theoretical predictions. In the nonadiabatic regime our measurements of the 85Rb molecule conversion efficiency follow a Landau-Zener model. PMID- 15903899 TI - Strongly inhibited transport of a degenerate 1D Bose gas in a lattice. AB - We report the observation of strongly damped dipole oscillations of a quantum degenerate 1D atomic Bose gas in a combined harmonic and optical lattice potential. Damping is significant for very shallow axial lattices (0.25 photon recoil energies), and increases dramatically with increasing lattice depth, such that the gas becomes nearly immobile for times an order of magnitude longer than the single-particle tunneling time. Surprisingly, we see no broadening of the atomic quasimomentum distribution after damped motion. Recent theoretical work suggests that quantum fluctuations can strongly damp dipole oscillations of a 1D atomic Bose gas, providing a possible explanation for our observations. PMID- 15903900 TI - Physical limits of heat-bath algorithmic cooling. AB - Simultaneous near-certain preparation of qubits (quantum bits) in their ground states is a key hurdle in quantum computing proposals as varied as liquid-state NMR and ion traps. "Closed-system" cooling mechanisms are of limited applicability due to the need for a continual supply of ancillas for fault tolerance, and to the high initial temperatures of some systems. "Open-system" mechanisms are therefore required. We describe a new, efficient initialization procedure for such open systems. With this procedure, an n-qubit device that is originally maximally mixed, but is in contact with a heat bath of bias epsilon>>2(-n), can be almost perfectly initialized. This performance is optimal due to a newly discovered threshold effect: for bias epsilon<<2(-n) no cooling procedure can, even in principle (running indefinitely without any decoherence), significantly initialize even a single qubit. PMID- 15903901 TI - Coexistence of black holes and a long-range scalar field in cosmology. AB - The exactly solvable scalar hairy black hole model (originated from the modern high-energy theory) is proposed. It turns out that the existence of black holes is strongly correlated to global scalar field, in a sense that they mutually impose bounds upon their physical parameters like the black hole mass (lower bound) or the cosmological constant (upper bound). We consider the same model also as a cosmological one and show that it agrees with recent experimental data; additionally, it provides a unified quintessence-like description of dark energy and dark matter. PMID- 15903902 TI - Parametric instabilities and their control in advanced interferometer gravitational-wave detectors. AB - A detailed simulation of Advanced LIGO test mass optical cavities shows that parametric instabilities will excite 7 acoustic modes in each fused silica test mass, with parametric gain R up to 7 and only 1 acoustic mode with R approximately 2 for alternative sapphire test masses. Fine-tuning of the test mass radii of curvature causes the instabilities to sweep through various modes with R as high as approximately 2000. Sapphire test mass cavities can be tuned to completely eliminate instabilities using thermal g-factor tuning. In the case of fused silica test mass, instabilities can be minimized but not eliminated. PMID- 15903904 TI - Detecting extra dimensions with gravity-wave spectroscopy: the black-string brane world. AB - Using the black string between two branes as a model of a brane-world black hole, we compute the gravity-wave perturbations and identify the features arising from the additional polarizations of the graviton. The standard four-dimensional gravitational wave signal acquires late-time oscillations due to massive modes of the graviton. The Fourier transform of these oscillations shows a series of spikes associated with the masses of the Kaluza-Klein modes, providing in principle a spectroscopic signature of extra dimensions. PMID- 15903903 TI - First results from the CERN axion solar telescope. AB - Hypothetical axionlike particles with a two-photon interaction would be produced in the sun by the Primakoff process. In a laboratory magnetic field ("axion helioscope"), they would be transformed into x-rays with energies of a few keV. Using a decommissioned Large Hadron Collider test magnet, the CERN Axion Solar Telescope ran for about 6 months during 2003. The first results from the analysis of these data are presented here. No signal above background was observed, implying an upper limit to the axion-photon coupling g(agamma)<1.16x10(-10) GeV-1 at 95% C.L. for m(a) less, similar 0.02 eV. This limit, assumption-free, is comparable to the limit from stellar energy-loss arguments and considerably more restrictive than any previous experiment over a broad range of axion masses. PMID- 15903905 TI - Study of B0-->rho+/- pi-/+ time-dependent CP violation at Belle. AB - We present a time-dependent analysis of CP violation in B0-->rho(+/-)pi(-/+) decays based on a 140 fb(-1) data sample collected at the Upsilon(4S) resonance with the Belle detector at KEKB. We obtain the charge asymmetry A(rhopi)(CP)= 0.16+/-0.10(stat)+/-0.02(syst). An unbinned maximum-likelihood fit to the Deltat distributions yields C(rhopi)=0.25+/-0.17(stat)+0.02-0.06(syst), DeltaC(rhopi)=0.38+/-0.18(stat)+0.02-0.04(syst), S(rhopi)=-0.28+/-0.23(stat)+0.10 0.08(syst), and DeltaS(rhopi)=-0.30+/-0.24(stat)+/-0.09(syst). The direct CP violation parameters for B-->rho(+)pi(-) and B-->rho(-)pi(+) decays are A(+ )(rhopi)=-0.02+/-0.16(stat)+0.05-0.02(syst) and A(-+)(rhopi)=-0.53+/ 0.29(stat)+0.09-0.04(syst). PMID- 15903908 TI - Correlated emission of hadrons from recombination of correlated partons. AB - We discuss different sources of hadron correlations in relativistic heavy ion collisions. We show that correlations among partons in a quasithermal medium can lead to the correlated emission of hadrons by quark recombination and argue that this mechanism offers a plausible explanation for the dihadron correlations in the few GeV/c momentum range observed in Au+Au collisions at the BNL Relativistic Heavy Ion Collider. PMID- 15903907 TI - Observation of an isotriplet of excited charmed baryons decaying to lambda+c pi. AB - We report the observation of an isotriplet of excited charmed baryons, decaying into Lambda(+)(c)pi(-), Lambda(+)(c)pi(0), and Lambda(+)(c)pi(+). We measure the mass differences M(Lambda(+)(c)pi)-M(Lambda(+)(c)) and widths to be 515.4(+3.2+2.1)(-3.1-6.0) MeV/c(2), 61(+18+22)(-13-13) MeV for the neutral state; 505.4(+5.8+12.4)(-4.6-2.0) MeV/c(2), 62(+37+52)(-23-38) MeV for the charged state; and 514.5(+3.4+2.8)(-3.1-4.9) MeV/c(2), 75(+18+12)(-13-11) MeV for the doubly charged state, where the uncertainties are statistical and systematic, respectively. These results are obtained from a 281 fb(-1) data sample collected with the Belle detector near the Upsilon(4S) resonance, at the KEKB asymmetric energy e(+)e(-) collider. PMID- 15903911 TI - B(E2) upward arrow measurements for radioactive neutron-rich ge isotopes: reaching the N=50 closed shell. AB - The B(E2;0(+)(1)-->2(+)(1)) values for the radioactive neutron-rich germanium isotopes (78,80)Ge and the closed neutron shell nucleus 82Ge were measured at the HRIBF using Coulomb excitation in inverse kinematics. These data allow a study of the systematic trend between the subshell closures at N=40 and 50. The B(E2) behavior approaching N=50 is similar to the trend observed for heavier isotopic chains. A comparison of the experimental results with a shell model calculation demonstrates persistence of the N=50 shell gap and a strong sensitivity of the B(E2) values to the effective interaction. PMID- 15903910 TI - Energy dependence of elliptic flow over a large pseudorapidity range in Au+Au collisions at the BNL relativistic heavy ion collider. AB - This Letter describes the measurement of the energy dependence of elliptic flow for charged particles in Au+Au collisions using the PHOBOS detector at the Relativistic Heavy Ion Collider. Data taken at collision energies of square root of s(NN)=19.6, 62.4, 130, and 200 GeV are shown over a wide range in pseudorapidity. These results, when plotted as a function of eta(')=|eta| y(beam), scale with approximate linearity throughout eta('), implying no sharp changes in the dynamics of particle production as a function of pseudorapidity or increasing beam energy. PMID- 15903912 TI - Direct observation of the ... 4+-to-2+ ... [corrected] gamma transition in 8Be. AB - Electromagnetic transition rates provide an important test for nuclear structure models. The nucleus 8Be exhibits a pronounced alpha-cluster structure and is a building block for more complex cluster nuclei. Here we report on the first observation of the gamma transition between the 4(+) and 2(+) states of 8Be in the 4He+4He reaction. The measured on-resonance cross section of 165+/-54 nb leads to a B(E2) of25+/-8e(2) fm(4), in good agreement with alpha-cluster models and sophisticated ab initio structure calculations. PMID- 15903913 TI - Role of the Pauli principle in collective-model coupled-channel calculations. AB - A multichannel algebraic scattering theory, to find solutions of coupled-channel scattering problems with interactions determined by collective models, has been structured to ensure that the Pauli principle is not violated. By tracking the results in the zero coupling limit, a correct interpretation of the subthreshold and resonant spectra of the compound system can be made. As an example, the neutron-12C system is studied defining properties of 13C to 10 MeV excitation. Accounting for the Pauli principle in collective coupled-channels models is crucial to the outcome. PMID- 15903914 TI - NMR detection with an atomic magnetometer. AB - We demonstrate detection of NMR signals using a noncryogenic atomic magnetometer and describe several novel applications of this technique. A nuclear spin precession signal from water is detected using a spin-exchange-relaxation-free potassium magnetometer. We also demonstrate detection of less than 10(13) 129Xe atoms whose NMR signal is enhanced by a factor of 540 due to Fermi-contact interaction with K atoms. The possibility of using a multichannel atomic magnetometer for fast 3D magnetic resonance imaging is also discussed. PMID- 15903915 TI - Zeeman relaxation of CaF in low-temperature collisions with helium. AB - The collision-induced Zeeman relaxation rate for collisions of CaF X2Sigma(v('')=0) with 3He is measured to be Gamma(Z)=(7.7+5.4/-2.5)x10(-15) cm(3)/s at 2 K. This rate is a direct measurement of the influence of spin rotation coupling on Zeeman relaxation in the first rotational level of CaF. The relationship of this rate to known molecular constants is consistent with recent theory of cold molecular collisions and outlines the (2)Sigma molecules conducive to magnetic trapping. PMID- 15903909 TI - Deuteron and antideuteron production in Au+Au collisions at square root of s(NN)=200 GeV. AB - The production of deuterons and antideuterons in the transverse momentum range 1.1D0pi(+) (and charge conjugate) decays with the D0 decaying to K-pi(+), K-K+, and pi(-)pi(+), corresponding to 123 pb(-1) of data collected by the Collider Detector at Fermilab II experiment at the Fermilab Tevatron collider. No significant direct CP violation is observed. We measure Gamma(D0-->K-K+)/Gamma(D0-->K pi(+))=0.0992+/-0.0011+/-0.0012, Gamma(D0-->pi(-)pi(+))/Gamma(D0-->K-pi(+))=0.035 94+/-0.000 54+/-0.000 40, A(CP)(K-K+)=(2.0+/-1.2+/-0.6)%, and A(CP)(pi( )pi(+))=(1.0+/-1.3+/-0.6)%, where, in all cases, the first uncertainty is statistical and the second is systematic. PMID- 15903917 TI - Observation of Feshbach-like resonances in collisions between ultracold molecules. AB - We observe magnetically tuned collision resonances for ultracold Cs2 molecules stored in a CO2-laser trap. By magnetically levitating the molecules against gravity, we precisely measure their magnetic moment. We find an avoided level crossing which allows us to transfer the molecules into another state. In the new state, two Feshbach-like collision resonances show up as strong inelastic loss features. We interpret these resonances as being induced by Cs4 bound states near the molecular scattering continuum. The tunability of the interactions between molecules opens up novel applications such as controlled chemical reactions and synthesis of ultracold complex molecules. PMID- 15903918 TI - Quantum imaging of nonlocal spatial correlations induced by orbital angular momentum. AB - Through scanned coincidence counting, we probe the quantum image produced by parametric down-conversion with a pump-beam carrying orbital angular momentum. Nonlocal spatial correlations are manifested through splitting of the coincidence spot into two. PMID- 15903919 TI - ac Stark shift and dephasing of a superconducting qubit strongly coupled to a cavity field. AB - We have performed spectroscopy of a superconducting charge qubit coupled nonresonantly to a single mode of an on-chip resonator. The strong coupling induces a large ac Stark shift in the energy levels of both the qubit and the resonator. The dispersive shift of the resonator frequency is used to nondestructively determine the qubit state. Photon shot noise in the measurement field induces qubit level fluctuations leading to dephasing which is characteristic for the measurement backaction. A crossover in line shape with measurement power is observed and theoretically explained. For weak measurement a long intrinsic dephasing time of T2>200 ns of the qubit is found. PMID- 15903920 TI - Direct observation of Bloch harmonics and negative phase velocity in photonic crystal waveguides. AB - The eigenfield distribution and the band structure of a photonic crystal waveguide have been measured with a phase-sensitive near-field scanning optical microscope. Bloch modes, which consist of more than one spatial frequency, are visualized in the waveguide. In the band structure, multiple Brillouin zones due to zone folding are observed, in which positive and negative dispersion is seen. The negative slopes are shown to correspond to a negative phase velocity but a positive group velocity. The lateral mode profile for modes separated by one reciprocal lattice vector is found to be different. PMID- 15903921 TI - Controlling high harmonic generation with molecular wave packets. AB - We show that, by controlling the alignment of molecules, we can influence the high harmonic generation process. We observed strong intensity modulation and spectral shaping of high harmonics produced with a rotational wave packet in a low-density gas of N2 or O2. In N2, where the highest occupied molecular orbital (HOMO) has sigma(g) symmetry, the maximum signal occurs when the molecules are aligned along the laser polarization while the minimum occurs when it is perpendicular. In O2, where the HOMO has pi(g) symmetry, the harmonics are enhanced when the molecules are aligned around 45 degrees to the laser polarization. The symmetry of the molecular orbital can be read by harmonics. Molecular wave packets offer a means of shaping attosecond pulses. PMID- 15903922 TI - Cavitation in a lubrication flow between a moving sphere and a boundary. AB - A heavy sphere is free to move inside a rotating horizontal cylinder filled with viscous liquid. The steady motion is essentially Stokesian, and the sphere rotates at a fixed location with a lubrication layer between the ball and the wall. The symmetry of the flow field suggests there will be no force to balance the normal component of the ball's weight. However, we show that a normal force can arise when a cavitation bubble is present. The bubble size was measured as a function of the cylinder rotation rate and agrees well with a model which uses the force and torque balances on the sphere. PMID- 15903923 TI - Exotic statistics of leapfrogging vortex rings. AB - The leapfrogging motion of vortex rings is a three-dimensional version of the motion that in two dimensions leads to exotic exchange statistics. The statistical phase factor can be computed using the hydrodynamical Euler equation, which suggests that three-dimensional exotic exchange statistics is a common property of vortex rings in a variety of quantum liquids and gases. Potential applications range from helium superfluids to Bose-Einstein condensed alkali gases, metallic hydrogen in its liquid phases, and maybe even nuclear matter in extreme conditions. PMID- 15903924 TI - Ionospheric modification at twice the electron cyclotron frequency. AB - In 2004, a new transmission band was added to the HAARP high-frequency ionospheric modification facility that encompasses the second electron cyclotron harmonic at altitudes between approximately 220 and 330 km. Initial observations indicate that greatly enhanced airglow occurs whenever the transmission frequency approximately matches the second electron cyclotron harmonic at the height of the upper hybrid resonance. This is the reverse of what happens at higher electron cyclotron harmonics. The measured optical emissions confirm the presence of accelerated electrons in the plasma. PMID- 15903925 TI - Effect of parallel flows and toroidicity on cross-field transport of pellet ablation matter in tokamak plasmas. AB - The first complete set of time-dependent equations describing the cross-field drift of ionized pellet ablation matter in tokamak plasma caused by polarization in the nonuniform magnetic field has been developed and solved numerically. Important new features impacting the drift dynamics have been identified, including the effect of pressure profile variations in the tokamak plasma, curvature drive by near-sonic field-aligned (parallel) flows, and the rotational transform of the magnetic field lines, and are considered from the viewpoint of the parallel vorticity equation. These new features are necessary to obtain favorable quantitative agreement between theory and experimental fuel deposition profiles for both inner and outer wall launched pellet injection cases on the DIII-D tokamak. PMID- 15903926 TI - Evolution of the current density profile associated with magnetic island formation in JT-60U. AB - Evolution of the current density profile associated with magnetic island formation of an m/n=2/1 tearing mode was measured using a motional Stark effect (MSE) diagnostic for the first time in the JT-60U tokamak. With the island growth, the current density profile turned flat at the radial region of the island, followed by an appearance of a hollow structure. As the island shrank, the flat region became narrower, and it finally diminished after the disappearance of the island. The fluctuation of the local poloidal magnetic field from MSE showed a strong correlation with a slow island rotation. This indicates that the observed deformation in the current density profile is localized at the island O point. PMID- 15903927 TI - Modal focusing effect of positive and negative ions by a three-dimensional plasma sheath lens. AB - A complex focusing effect of positive and negative ions caused by the sheath forming to biased electrodes that interface insulators has been found by solving in three dimensions the potential distribution and ion kinetics within the sheath. Thus, intrinsically correlated with the sheath shape, certain electrical charges are focused on the surface, forming well defined patterns named modal lines and modal spots. Their superposition to the previously reported discrete focusing leads to a total flux that represents a "fingerprint" of the entire sheath on the electrode surface. The ion flux pattern is developed experimentally on the surface of square and octagonal electrodes exposed to Ar/SF(6) and CF4 plasmas. Present results are of high potential importance for fundamental studies concerning sheath formation and charge kinetics and also in a wide range of plasma applications. PMID- 15903928 TI - Delocalized nature of the E'delta center in amorphous silicon dioxide. AB - We report an experimental study by electron paramagnetic resonance (EPR) of E(')(delta) point defect induced by gamma-ray irradiation in amorphous SiO2. We obtained an estimation of the intensity of the 10 mT doublet characterizing the EPR spectrum of such a defect arising from hyperfine interaction of the unpaired electron with a 29Si (I=1/2) nucleus. Moreover, determining the intensity ratio between this hyperfine doublet and the main resonance line of E(')(delta) center, we pointed out that the unpaired electron wave function of this center is actually delocalized over four nearly equivalent silicon atoms. PMID- 15903929 TI - Compact and dissociated dislocations in aluminum: implications for deformation. AB - Atomistic simulations, confirmed by electron microscopy, show that dislocations in aluminum can have compact or dissociated cores. The calculated minimum stress (sigma(P)) required to move an edge dislocation is approximately 20 times smaller for dissociated than for equivalent compact dislocations. This contradicts the well accepted generalized stacking fault energy paradigm that predicts similar sigma(P) values for both configurations. Additionally, Frank's rule and the Schmid law are also violated because dislocation core energies become important. These results may help settle a 50-year-old puzzle regarding the magnitude of sigma(P) in face-centered-cubic metals, and provide new insights into the deformation of ultra-fine-grained metals. PMID- 15903930 TI - Theoretical evidence for a reentrant phase diagram in ortho-para mixtures of solid H2 at high pressure. AB - We develop a multiorder parameter mean-field formalism for systems of coupled quantum rotors. The scheme is developed to account for systems where ortho-para distinction is valid. We apply our formalism to solid H2 and D2. We find an anomalous reentrant orientational phase transition for both systems at thermal equilibrium. The correlation functions of the order parameter indicate short range order at low temperatures. As the temperature is increased the correlation increases along the phase boundary. We also find that even extremely small odd-J concentrations (1%) can trigger short-range orientational ordering. PMID- 15903931 TI - Shock temperature measurement using neutron resonance spectroscopy. AB - We report a direct measurement of temperature in a shocked metal using Doppler broadening of neutron resonances. The 21.1-eV resonance in 182W was used to measure the temperature in molybdenum shocked to approximately 63 GPa. An explosively launched aluminum flyer produced a planar shock in a molybdenum target that contained a 1-mm thick layer doped with 1.7 at. %(182)W. A single neutron pulse, containing resonant neutrons of less than 1 mus duration, probed the shocked material. Fits to the neutron time-of-flight data were used to determine the temperature of the shocked molybdenum. PMID- 15903932 TI - Consecutive rotation of crystallographic orientation in lateral growth. AB - A consecutive rotation of crystallographic orientation has been observed in lateral crystallization of NH4Cl on a glass substrate, which induces a periodic distribution of faceted and roughened regions on the surface of a crystallite aggregate. Experimental observation indicates that this phenomenon derives from the asymmetric surface energies at the growth front, which deform the nascent nucleus and tilt the crystallographic orientation in the nucleation-mediated layered growth. We suggest that this effect is significant for a class of lateral growth where nucleation plays a dominate role. PMID- 15903933 TI - Nature of amorphous polymorphism of water. AB - We report elastic and inelastic neutron scattering experiments on different amorphous ice modifications. It is shown that an amorphous structure (HDA') indiscernible from the high-density phase (HDA), obtained by compression of crystalline ice, can be formed from the very high-density phase (vHDA) as an intermediate stage of the transition of vHDA into its low-density modification (LDA'). Both HDA and HDA' exhibit comparable small-angle scattering signals characterizing them as structures heterogeneous on a length scale of a few nanometers. The homogeneous structures are the initial and final transition stages vHDA and LDA', respectively. Despite their apparent structural identity on a local scale, HDA and HDA' differ in their transition kinetics explored by in situ experiments. The activation energy of the vHDA-to-LDA' transition is at least 20 kJ/mol higher than the activation energy of the HDA-to-LDA transition. PMID- 15903934 TI - X-ray diffraction study of liquid Cs up to 9.8 GPa. AB - We describe an x-ray diffraction study of liquid Cs at high pressure and temperature conducted in order to characterize the structural changes associated with the complex melting curve and phase transitions observed in the solid phases. At 3.9 GPa we observe a discontinuity in the density of the liquid accompanied by a decrease in the coordination number from about 12 to 8, which marks a change to a nonsimple liquid. The specific volume of liquid Cs, combined with structural analysis of the diffraction data, strongly suggest the existence of dsp(3) electronic hybridization above 3.9 GPa, similar to that reported on compression in the crystalline phase. PMID- 15903935 TI - Dynamic ionization of water under extreme conditions. AB - Raman spectroscopy in a laser heated diamond anvil cell and first principles molecular dynamics simulations have been used to study water in the temperature range 300 to 1500 K and at pressures to 56 GPa. We find a substantial decrease in the intensity of the O-H stretch mode in the liquid phase with pressure, and a change in slope of the melting line at 47 GPa and 1000 K. Consistent with these observations, theoretical calculations show that water beyond 50 GPa is "dynamically ionized" in that it consists of very short-lived (<10 fs) H2O, H3O+, and OH- species, and also that the mobility of the oxygen ions decreases abruptly with pressure, while hydrogen ions remain very mobile. We suggest that this regime corresponds to a superionic state. PMID- 15903936 TI - Picosecond x-ray studies of coherent folded acoustic phonons in a multiple quantum well. AB - Coherent folded acoustic phonons in a multilayered GaSb/InAs epitaxial heterostructure were generated by femtosecond laser pulses and studied by means of ultrafast x-ray diffraction. Coherent phonons excited simultaneously in the fundamental acoustic branch and the first back-folded branch were detected. This represents the first clear evidence for phonon branch folding based directly on the atomic motion to which x-ray diffraction is sensitive. From a comparison of the measured phonon-modulated x-ray reflectivity with simulations, evidence was found for a reduction of the laser penetration depth. This reduction can be explained by the self-modulation of the refractive index due to photogenerated free carriers. PMID- 15903937 TI - Fracture of brittle metallic glasses: brittleness or plasticity. AB - We report a brittle Mg-based bulk metallic glass which approaches the ideal brittle behavior. However, a dimple structure is observed at the fracture surface by high resolution scanning electron microscopy, indicating some type of "ductile" fracture mechanism in this very brittle glass. We also show, from the available data, a clear correlation between the fracture toughness and plastic process zone size for various glasses. The results indicate that the fracture in brittle metallic glassy materials might also proceed through the local softening mechanism but at different length scales. PMID- 15903938 TI - Formation of odd-numbered clusters of CO2 adsorbed on nanotube bundles. AB - Simulations show that CO2 adsorbed in the groove sites of carbon nanotubes displays unique quasi-one-dimensional behavior. Clusters containing only odd numbers of molecules are formed at finite CO2 coverages and low temperatures. The molecules are orientationally ordered with respect to the nanotube groove axis and azimuthally ordered in the plane perpendicular to the groove axis. This ordering is a result of a delicate balance between solid-fluid and fluid-fluid forces; the CO2 quadrupole plays a critical role in the cluster formation and orientational ordering. PMID- 15903939 TI - Anomalous diffusion profiles of Ag in CdTe due to chemical self-diffusion. AB - Anomalous diffusion profiles of Ag in single crystalline CdTe were observed using the radiotracer 111Ag. The diffusion anneals were performed at 800 K under Cd or Te vapor and in a vacuum for different Ag concentrations. The measured Ag profiles directly reflect the distribution of the self-interstitials and vacancies of the Cd sublattice and are the result of chemical self-diffusion which describes the variation of the deviation from stoichiometry of the binary crystal as a function of depth and time. PMID- 15903940 TI - Jahn-Teller stabilization of a "polar" metal oxide surface: Fe3O4(001). AB - Using ab initio thermodynamics we compile a phase diagram for the surface of Fe3O4(001) as a function of temperature and oxygen pressures. A hitherto ignored polar termination with octahedral iron and oxygen forming a wavelike structure along the [110] direction is identified as the lowest energy configuration over a broad range of oxygen gas-phase conditions. This novel geometry is confirmed via x-ray diffraction analysis. The stabilization of the Fe3O4(001) surface goes together with dramatic changes in the electronic and magnetic properties, e.g., a half metal to metal transition. PMID- 15903941 TI - Atom transfer and single-adatom contacts. AB - The point contact of a tunnel tip approaching towards Ag(111) and Cu(111) surfaces is investigated with a low temperature scanning tunneling microscope. A sharp jump to contact, random in nature, is observed in the conductance. After point contact, the tip-apex atom is transferred to the surface, indicating that a one-atom contact is formed during the approach. In sharp contrast, the conductance over single silver and copper adatoms exhibits a smooth and reproducible transition from tunneling to contact regime. Numerical simulations show that this is a consequence of the additional dipolar bonding between the adatom and the surface atoms. PMID- 15903942 TI - Very low friction state of a dodecane film confined between mica surfaces. AB - The existence of a very low friction state of a lubrication film is demonstrated in nonequilibrium molecular dynamics simulations of a six-layer dodecane film between mica walls. We argue that this low friction state is thermodynamically stable with respect to the well documented high friction film, the latter being a metastable state. These results are in striking accord with the recent report of Zhu and Granick [Phys. Rev. Lett. 93, 096101 (2004).]. The extreme low friction is the result, not of wall slip, but of layer sliding throughout the film, a mechanism similar to solid lubrication. PMID- 15903943 TI - Electric field effect in diluted magnetic insulator anatase Co: TiO2. AB - An external electric field induced reversible modulation of a room temperature magnetic moment and coercive field is achieved in an epitaxial and insulating thin film of dilutely cobalt-doped anatase TiO2. This first demonstration of an electric field effect in any oxide-based diluted ferromagnet is realized in a high quality epitaxial heterostructure of PbZr(0.2)Ti(0.8)O(3)/Co: TiO(2)/SrRuO(3) grown on (001) LaAlO3. The observed effect, which is about 15% in strength in a given heterostructure, can be modulated over several cycles. Possible mechanisms for electric field induced modulation of insulating ferromagnetism are discussed. PMID- 15903944 TI - Space-charge limited photocurrent. AB - In 1971 Goodman and Rose predicted the occurrence of a fundamental electrostatic limit for the photocurrent in semiconductors at high light intensities. Blends of conjugated polymers and fullerenes are an ideal model system to observe this space-charge limit experimentally, since they combine an unbalanced charge transport, long lifetimes, high charge carrier generation efficiencies, and low mobility of the slowest charge carrier. The experimental photocurrents reveal all the characteristics of a space-charge limited photocurrent: a one-half power dependence on voltage, a three-quarter power dependence on light intensity, and a one-half power scaling of the voltage at which the photocurrent switches into full saturation with light intensity. PMID- 15903945 TI - Imaging electron interferometer. AB - An imaging interferometer was created in a two-dimensional electron gas by reflecting electron waves emitted from a quantum point contact with a circular mirror. Images of electron flow obtained with a scanning probe microscope at liquid He temperatures show interference fringes when the mirror is energized. A quantum phase shifter was created by moving the mirror via its gate voltage, and an interferometric spectrometer can be formed by sweeping the tip over many wavelengths. Experiments and theory demonstrate that the interference signal is robust against thermal averaging. PMID- 15903946 TI - Spin manipulation of free two-dimensional electrons in Si/SiGe quantum wells. AB - Understanding the mechanisms controlling the spin coherence of electrons in semiconductors is essential for designing structures for quantum computing applications. Using a pulsed electron paramagnetic resonance spectrometer, we measure spin echoes and deduce a spin coherence time (T2) of up to 3 mus for an ensemble of free two-dimensional electrons confined in a Si/SiGe quantum well. The decoherence can be understood in terms of momentum scattering causing fluctuating effective Rashba fields. Further confining the electrons into a nondegenerate (other than spin) ground state of a quantum dot can be expected to eliminate this decoherence mechanism. PMID- 15903947 TI - Interplay between the mesoscopic Stoner and Kondo effects in quantum dots. AB - We consider electrons confined to a quantum dot interacting antiferromagnetically with a spin-1 / 2 Kondo impurity. The electrons also interact among themselves ferromagnetically with a dimensionless coupling J , where J =1 denotes the bulk Stoner transition. We show that as J approaches 1 there is a regime with enhanced Kondo correlations, followed by one where the Kondo effect is destroyed and impurity is spin polarized opposite to the dot electrons. The most striking signature of the first, Stoner-enhanced Kondo regime is that a Zeeman field increases the Kondo scale, in contrast to the case for noninteracting dot electrons. Implications for experiments are discussed. PMID- 15903948 TI - Electronic structure and dynamics of quantum-well states in thin Yb metal films. AB - Quantum-well states above the Fermi energy in thin Yb(111) metal films deposited on a W(110) single crystal were studied by low-temperature scanning tunneling spectroscopy. These states are laterally highly localized and give rise to sharp peaks in the tunneling spectra. A quantitative analysis of the spectra yields the bulk-band dispersion in the Gamma-L direction as well as quasiparticle lifetimes. The quadratic energy dependence of the lifetimes is in quantitative agreement with Fermi-liquid theory. PMID- 15903949 TI - Phonon-induced optical superlattice. AB - We demonstrate the formation of a dynamic optical superlattice through the modulation of a semiconductor microcavity by stimulated acoustic phonons. The high coherent phonon population produces a folded optical dispersion relation with well-defined energy gaps and renormalized energy levels, which are accessed using reflection and diffraction experiments. PMID- 15903950 TI - Muon-spin-rotation measurements of the penetration depth of the infinite-layer electron-doped Sr0.9La0.1CuO2 cuprate superconductor. AB - Muon-spin-rotation (muSR) measurements of the in-plane penetration depth lambda(ab) have been performed in the infinite-layer electron-doped Sr0.9La0.1CuO2 high-T(c) superconductor (HTS). Absence of the magnetic rare-earth ions in this compound allowed us to measure for the first time the absolute value of lambda(ab)(0) in electron-doped HTSs using muSR. We found lambda(ab)(0)=116(2) nm. The zero-temperature depolarization rate sigma(0) proportional, variant 1/lambda(2)(ab)(0)=4.6(1) micros(-1) is more than 4 times higher than expected from the Uemura line. Therefore, this electron-doped HTS does not follow the Uemura relation found for hole-doped HTSs. PMID- 15903951 TI - Low- and high-frequency noise from coherent two-level systems. AB - Recent experiments indicate a connection between the low- and high-frequency noises affecting superconducting quantum systems. We explore the possibilities that both noises can be produced by one ensemble of microscopic modes, made up, e.g., by sufficiently coherent two-level systems (TLS's). This implies a relation between the noise power in different frequency domains, which depends on the distribution of the parameters of the TLS's. We show that a distribution, natural for tunneling TLS's, with a log-uniform distribution in the tunnel splitting and linear distribution in the bias, accounts for experimental observations. PMID- 15903952 TI - Pairing and superconductivity driven by strong quasiparticle renormalization in two-dimensional organic charge transfer salts. AB - We introduce and analyze a variational wave function for quasi-two-dimensional kappa-(ET)(2) organic salts containing strong local and nonlocal correlation effects. We find an unconventional superconducting ground state for intermediate charge carrier interaction, sandwiched between a conventional metal at weak coupling and a spin liquid at larger coupling. Most remarkably, the excitation spectrum is dramatically renormalized and is found to be the driving force for the formation of the unusual superconducting state. PMID- 15903953 TI - Monte Carlo simulations of ferromagnetism in p-Cd1-xMnxTe quantum wells. AB - Monte Carlo simulations, in which the Schrodinger equation is solved at each Monte Carlo sweep, are employed to assess the influence of magnetization fluctuations, short-range antiferromagnetic interactions, disorder, magnetic polaron formation, and spin-Peierls instability on the carrier-mediated Ising ferromagnetism in two-dimensional electronic systems. The determined critical temperature and hysteresis are affected in a nontrivial way by the antiferromagnetic interactions. The findings explain striking experimental results for modulation-doped p-Cd1-xMnxTe quantum wells. PMID- 15903954 TI - Large tunneling anisotropic magnetoresistance in (Ga,Mn)As nanoconstrictions. AB - We report a large tunneling anisotropic magnetoresistance (TAMR) in (Ga,Mn)As lateral nanoconstrictions. Unlike previously reported tunneling magnetoresistance effects in nanocontacts, the TAMR does not require noncollinear magnetization on either side of the constriction. The nature of the effect is established by a direct comparison of its phenomenology with that of normal anisotropic magnetoresistance (AMR) measured in the same lateral geometry. The direct link we establish between the TAMR and AMR indicates that TAMR may be observable in other materials showing room temperature AMR and demonstrates that the physics of nanoconstriction magnetoresistive devices can be much richer than previously thought. PMID- 15903955 TI - Ballistic anisotropic magnetoresistance. AB - Electronic transport in ferromagnetic ballistic conductors is predicted to exhibit ballistic anisotropic magnetoresistance-a change in the ballistic conductance with the direction of magnetization. This phenomenon originates from the effect of the spin-orbit interaction on the electronic band structure which leads to a change in the number of bands crossing the Fermi energy when the magnetization direction changes. We illustrate the significance of this phenomenon by performing ab initio calculations of the ballistic conductance in ferromagnetic Ni and Fe nanowires which display a sizable ballistic anisotropic magnetoresistance when magnetization changes direction from parallel to perpendicular to the wire axis. PMID- 15903956 TI - Hydrogen-mediated spin-spin interaction in ZnCoO. AB - The effect of hydrogen impurities on the electronic and magnetic properties of ZnO-based diluted magnetic semiconductors is examined through first-principles pseudopotential calculations. We suggest that interstitial H can mediate a strong short-ranged ferromagnetic spin-spin interaction between neighboring magnetic impurities through the formation of a bridge bond. Results based on first principles total-energy calculations and Monte Carlo simulations indicate that such H-mediated spin-spin interactions can lead to high temperature ferromagnetism. PMID- 15903957 TI - Micromagnetic dissipation, dispersion, and mode conversion in thin permalloy platelets. AB - Micron-sized ferromagnetic Permalloy disks exhibiting an in-plane ferromagnetic vortex structure are excited by a fast rise time perpendicular magnetic field pulse and their modal structure is analyzed. We find azimuthal and axial modes. By a Fourier filtering technique we can separate and analyze the time dependence of individual modes. Analysis of the experimental data demonstrates that the azimuthal modes damp more quickly than the axial modes. We interpret these results as mode conversion from low-frequency azimuthal modes to the fundamental mode which is higher in frequency, i.e., mode-mode coupling in a system with a single Landau-Lifshitz-Gilbert phenomenological damping constant alpha. PMID- 15903958 TI - Magnetization switching and microwave oscillations in nanomagnets driven by spin polarized currents. AB - A novel theoretical approach to magnetization dynamics driven by spin-polarized currents is presented. Complete stability diagrams are obtained for the case where spin torques and external magnetic fields are simultaneously present. Quantitative predictions are made for the critical currents and fields inducing magnetization switching, for the amplitude and frequency of magnetization self oscillations, and for the conditions leading to hysteretic transitions between self-oscillations and stationary states. PMID- 15903959 TI - Zener tunneling of light waves in an optical superlattice. AB - We report on the observation of Zener tunneling of light waves in spectral and time-resolved transmission measurements, performed on an optical superlattice made of porous silicon. The structure was designed to have two photonic minibands, spaced by a narrow frequency gap. A gradient in the refractive index was introduced to create two optical Wannier-Stark ladders and, at a critical value of the optical gradient, tunneling between energy bands was observed in the form of an enhanced transmission peak and a characteristic time dependence of the transmission. PMID- 15903960 TI - Phonon-assisted excitonic recombination channels observed in DNA-wrapped carbon nanotubes using photoluminescence spectroscopy. AB - By using a sample of DNA-wrapped single-wall carbon nanotubes strongly enriched in the (6,5) nanotube, photoluminescence emissions observed at special excitation energy values were identified with specific mechanisms of phonon-assisted excitonic absorption and recombination processes associated with (6,5) nanotubes, including one-phonon, two-phonon, and some continuous-luminescence processes. Such detailed processes are not separately identified in three-dimensional semiconducting materials. A general theoretical framework is presented to interpret the experimentally observed phonon-assisted processes in terms of excitonic states. PMID- 15903961 TI - Single carbon nanotubes probed by photoluminescence excitation spectroscopy: the role of phonon-assisted transitions. AB - We study light absorption mechanisms in semiconducting carbon nanotubes using low temperature, single-nanotube photoluminescence excitation spectroscopy. In addition to purely electronic transitions, we observe several strong phonon assisted bands due to excitation of one or more phonon modes together with the first electronic state. In contrast with a small width of emission lines (sub-meV to a few meV), most of the photoluminescence excitation features have significant linewidths of tens of meV. All of these observations indicate very strong electron-phonon coupling that allows efficient excitation of electronic states via phonon-assisted processes and leads to ultrafast intraband relaxation due to inelastic electron-phonon scattering. PMID- 15903962 TI - Chain entanglement in thin freestanding polymer films. AB - When a thin glassy film is strained uniaxially, a shear deformation zone (SDZ) can be observed. The ratio of the thickness of the SDZ to that of the undeformed film is related to the maximum extension ratio, lambda, which depends on the entanglement molecular weight, M(e). We have measured lambda as a function of film thickness in strained freestanding films of polystyrene as a probe of M(e) in confinement. It is found that thin films stretch further than thick films before failure, consistent with the interpretation that polymers in thin films are less entangled than bulk polymers, thus the effective value of M(e) in thin films is significantly larger than that of the bulk. Our results are well described by a conceptually simple model based on the probability of finding intermolecular entanglements near an interface. PMID- 15903963 TI - Bagnold scaling, density plateau, and kinetic theory analysis of dense granular flow. AB - We investigate the bulk rheology of dense granular flow down a rough slope, where the density profile has been found to show a plateau except for the boundary layers in simulations [Silbert et al., Phys. Rev. E 64, 051302 (2001)]. It is demonstrated that both the Bagnold scaling and the framework of kinetic theory are applicable in the bulk, which allows us to extract the constitutive relations from simulation data. The detailed comparison of our data with the kinetic theory shows quantitative agreement for the normal and shear stresses, but there exists a slight discrepancy in the energy dissipation, which causes a rather large disagreement in the kinetic theory analysis of the flow. PMID- 15903964 TI - Picosecond thermometer in the amide I band of myoglobin. AB - The amide I and II bands in myoglobin show a heterogeneous temperature dependence, with bands at 6.17 and 6.43 microm which are more intense at low temperatures. The amide I band temperature dependence is on the long wavelength edge of the band, while the short wavelength side has almost no temperature dependence. We compare concepts of anharmonic solid-state crystal physics and chemical physics for the origins of these bands. We suggest that the long wavelength side is composed of those amino acids which hydrogen bond to the hydration shell of the protein, and that temperature dependent bands can be used to determine the time it takes vibrational energy to flow into the hydration shell. We determine that vibrational energy flow to the hydration shell from the amide I takes approximately 20 ps to occur. PMID- 15903965 TI - Unravelling the mechanism of RNA-polymerase forward motion by using mechanical force. AB - Polymerases form a class of enzymes that act as molecular motors as they move along their nucleic acid substrate during catalysis, incorporating nucleotide triphosphates at the end of the growing chain and consuming chemical energy. A debated issue is how the enzyme converts chemical energy into motion [J. Gelles and R. Landick, Cell 93, 13 (1998)]. In a single molecule assay, we studied how an opposing mechanical force affects the translocation rate of T7 RNA polymerase. Our measurements show that force acts as a competitive inhibitor of nucleotide binding. This result is interpreted in the context of possible models, and with respect to published crystal structures of T7 RNA polymerase. The transcribing complex appears to utilize only a small fraction of the energy of hydrolysis to perform mechanical work, with the remainder being converted to heat. PMID- 15903966 TI - Simulating biochemical networks at the particle level and in time and space: Green's function reaction dynamics. AB - We present a technique, called Green's function reaction dynamics (GFRD), for particle-based simulations of reaction-diffusion systems. GFRD uses a maximum time step such that only single particles or pairs of particles have to be considered. For these particles, the Smoluchowski equations are solved analytically using Green's functions, which are used to set up an event-driven algorithm. We apply the technique to a model of gene expression. Under biologically relevant conditions, GFRD is up to 5 orders of magnitude faster than conventional particle-based schemes. PMID- 15903967 TI - Frequency-locking phenomena of propagating wave fronts in reaction-diffusion systems. AB - We observe N:(N-1)(N>/=2) frequency-locking phenomena of propagating wave fronts when increasing the light intensity in a spatially extended system. The experiments were carried out using the light-sensitive form of the Belousov Zhabotinsky reaction with Ru(bpy)(2+)3 as a catalyst. By constructing a mapping function, the characteristic devil's staircase can be reproduced when plotting wave period versus light intensity, in agreement with the experimental data. PMID- 15903968 TI - Cell fates as high-dimensional attractor states of a complex gene regulatory network. AB - Cells in multicellular organisms switch between distinct cell fates, such as proliferation or differentiation into specialized cell types. Genome-wide gene regulatory networks govern this behavior. Theoretical studies of complex networks suggest that they can exhibit ordered (stable) dynamics, raising the possibility that cell fates may represent high-dimensional attractor states. We used gene expression profiling to show that trajectories of neutrophil differentiation converge to a common state from different directions of a 2773-dimensional gene expression state space, providing the first experimental evidence for a high dimensional stable attractor that represents a distinct cellular phenotype. PMID- 15903969 TI - Comment on "Crack street: the cycloidal wake of a cylinder tearing through a thin sheet". PMID- 15903971 TI - Comment on "Thermal glass transition beyond the Vogel-Fulcher-Tammann behavior for glass forming diglycidylether of bisphenol A". PMID- 15903973 TI - Time-reversal test for stochastic quantum dynamics. AB - The calculation of quantum dynamics is currently a central issue in theoretical physics, with diverse applications ranging from ultracold atomic Bose-Einstein condensates to condensed matter, biology, and even astrophysics. Here we demonstrate a conceptually simple method of determining the regime of validity of stochastic simulations of unitary quantum dynamics by employing a time-reversal test. We apply this test to a simulation of the evolution of a quantum anharmonic oscillator with up to 6.022x10(23) (Avogadro's number) of particles. This system is realizable as a Bose-Einstein condensate in an optical lattice, for which the time-reversal procedure could be implemented experimentally. PMID- 15903974 TI - Tuning indistinguishability in Bose-Einstein condensation and new atomic states. AB - The concept of indistinguishability is the key element in quantum statistics. But, are particles really either indistinguishable or distinguishable? Most works begin with the premise that all the particles, for example, in a quantum gas, are indistinguishable. Can we vary the degree of distinguishability in some controlled, continuous manner and see how it affects the behavior of the quantum gas, such as Bose-Einstein condensation (BEC)? We have found a complex parameter with a definite phase that does just that. As it deviates from zero, a gas mixture of originally indistinguishable bosons would divide into several distinct species that undergo BEC individually. Each species is found to be in a new type of atomic state whose spin structure adapts itself to the prevailing densities of the gases in the mixture. PMID- 15903975 TI - Lattice solitons in quasicondensates. AB - We analyze finite temperature effects in the generation of bright solitons in condensates in optical lattices. We show that even in the presence of strong phase fluctuations solitonic structures with a well defined phase profile can be created. We propose a novel family of variational functions which describe well the properties of these solitons and account for the nonlinear effects in the band structure. We discuss also the mobility and collisions of these localized wave packets. PMID- 15903976 TI - Resonant nonlinear quantum transport for a periodically kicked bose condensate. AB - Our realistic numerical results show that the fundamental and higher-order quantum resonances of the delta-kicked rotor are observable in state-of-the-art experiments with a Bose condensate in a shallow harmonic trap, kicked by a spatially periodic optical lattice. For stronger confinement, interaction-induced destruction of the resonant motion of the kicked harmonic oscillator is predicted. PMID- 15903977 TI - Experimental implementation of a concatenated quantum error-correcting code. AB - Concatenated coding provides a general strategy to achieve the desired level of noise protection in quantum information processing. We report the implementation of a concatenated quantum error-correcting code able to correct phase errors with a strong correlated component. The experiment was performed using liquid-state nuclear magnetic resonance techniques on a four spin subsystem of labeled crotonic acid. Our results show that concatenation between active and passive quantum error correction is a practical tool to handle realistic noise involving both independent and correlated errors. PMID- 15903978 TI - Equation of state of an interacting bose gas confined by a harmonic trap: the role of the "harmonic" pressure. AB - A gas of interacting atoms confined by a three dimensional anisotropic harmonic potential is studied. It is shown that there appear "new" thermodynamic variables instead of the usual pressure and volume: the latter is replaced by (the inverse of) the cube of the geometric average of the oscillator frequencies of the trap, and the former by the harmonic pressure responsible for the mechanical equilibrium of the fluid in the trap. We discuss the origin and physical meaning of these quantities and show that the equation of state of the gas is given in terms of these variables. The equation of state of a cold gas of interacting Bose atoms in the Hartree-Fock approximation is presented. We indicate how the harmonic pressure can be measured in current experiments. PMID- 15903979 TI - Derivation of the order parameter of the chiral Potts model. AB - We derive the order parameter of the chiral Potts model, using the method of Jimbo et al. The result agrees with previous conjectures. PMID- 15903980 TI - Gravitational-wave emission from rotating gravitational collapse in three dimensions. AB - We present the first three-dimensional (3D) calculations of the gravitational wave emission in the collapse of uniformly rotating stars to black holes. The initial models are polytropes which are dynamically unstable and near the mass shedding limit. The waveforms have been extracted using a gauge-invariant approach and reflect the properties of both the initial stellar models and of newly produced black holes, being in good qualitative agreement with those computed in previous 2D simulations. The wave amplitudes, however, are about 1 order of magnitude smaller, giving, for a source at 10 kpc, a signal-to-noise ratio S/N approximately 0.25 for LIGO-VIRGO and S/N less than or approximately equal 4 for LIGO II. PMID- 15903981 TI - Gamma rays from Kaluza-Klein dark matter. AB - A TeV gamma-ray signal from the direction of the Galactic center (GC) has been detected by the HESS experiment. Here, we investigate whether Kaluza-Klein (KK) dark matter annihilations near the GC can be the explanation. Including the contributions from internal bremsstrahlung as well as subsequent decays of quarks and tau leptons, we find a very flat gamma-ray spectrum which drops abruptly at the dark matter particle mass. For a KK mass of about 1 TeV, this gives a good fit to the HESS data below 1 TeV. A similar model, with gauge coupling roughly 3 times as large and a particle mass of about 10 TeV, would give both the correct relic density and a photon spectrum that fits the complete range of data. PMID- 15903982 TI - Helicity conservation in gauge boson scattering at high energy. AB - We remark that the high energy gauge boson scattering processes involving two body initial and final states satisfy certain selection rules described as helicity conservation of the gauge boson amplitudes (GBHC). These rules are valid at the Born level, as well as at the level of the leading and subleading 1-loop logarithmic corrections, in both the standard model and the minimal supersymmetric standard model (MSSM). A "fermionic equivalence" theorem is also proved, which suggests that GBHC is valid at all orders in the MSSM at sufficiently high energies, where the mass suppressed contributions are neglected. PMID- 15903983 TI - Brane worlds in collision. AB - We obtain an exact solution of the supergravity equations of motion in which the four-dimensional observed Universe is one of a number of colliding D3 branes in a Calabi-Yau background. The collision results in the ten-dimensional spacetime splitting into disconnected regions, bounded by curvature singularities. However, near the D3 branes the metric remains static during and after the collision. We also obtain a general class of solutions representing p-brane collisions in arbitrary dimensions, including one in which the universe ends with the mutual annihilation of a positive-tension and a negative-tension 3 brane. PMID- 15903985 TI - Dual Meissner effect and magnetic displacement currents. AB - The dual Meissner effect is observed without monopoles in quenched SU(2) QCD with Landau gauge fixing. Magnetic displacement currents that are time-dependent Abelian magnetic fields act as solenoidal currents squeezing Abelian electric fields. Monopoles are not always necessary for the dual Meissner effect. A mean field calculation suggests that the dual Meissner effect through the mass generation of the Abelian electric field is related to a gluon condensate A(a)(mu)A(a)(mu) not equal 0 of mass dimension 2. PMID- 15903986 TI - Anisotropic flow and jet quenching in ultrarelativistic U+U collisions. AB - Full-overlap U+U collisions provide significantly larger initial energy densities at comparable spatial deformation, and significantly larger deformation and volume at comparable energy density, than semi-central Au+Au collisions. We show quantitatively that this provides a long lever arm for studying the hydrodynamic behavior of elliptic flow in much larger and denser collision systems and the predicted nonlinear path-length dependence of radiative parton energy loss. PMID- 15903987 TI - Power-law tails from multiplicative noise. AB - We show that the well-known linear Langevin equation, modeling the Brownian motion and leading to a Gaussian stationary distribution of the corresponding Fokker-Planck equation, is changed by the smallest multiplicative noise. This leads to a power-law tail of the distribution for sufficiently large momenta. At finite ratio of the correlation strength for the multiplicative and the additive noises the stationary energy distribution becomes exactly the Tsallis distribution. PMID- 15903988 TI - p-sd Shell gap reduction in neutron-rich systems and cross-shell excitations in 20 O. AB - Excited states in 20O were populated in the reaction 10Be(14C,alpha) at Florida State University (FSU). Charged particles were detected with a particle telescope consisting of 4 annularly segmented Si surface barrier detectors and gamma radiation was detected with the FSU gamma detector array. Five new states were observed below 6 MeV from the alpha-gamma and alpha-gamma-gamma coincidence data. Shell model calculations suggest that most of the newly observed states are core excited 1p-1h excitations across the N=Z=8 shell gap. Comparisons between experimental data and calculations for the neutron-rich O and F isotopes imply a steady reduction of the p-sd shell gap as neutrons are added. PMID- 15903989 TI - Fine structure of the 1s3p 3PJ level in atomic 4He: theory and experiment. AB - The fine structure intervals in helium have been the focus of many theoretical and experimental studies in recent years with most of them concentrating on the 1s2p (3)P(J) levels. Here, we report on a theoretical calculation and an experimental determination of the 1s2p (3)P(J) fine structure intervals. The values from the theoretical calculation are 8113.730(6) and 658.801(6) MHz for the nu(01) and nu(12) intervals, respectively. The laser spectroscopic measurement reported here yields 8113.714(28) and 658.810(18) MHz for these intervals and is in excellent agreement with the theoretical calculation. Both, however, disagree significantly with the previous most precise experimental results. PMID- 15903990 TI - Geometry and spectrum of Casimir forces. AB - We present a new approach to the Helmholtz spectrum for arbitrarily shaped boundaries and general boundary conditions. We derive the boundary induced change of the density of states in terms of the free Green's function from which we obtain nonperturbative results for the Casimir interaction between rigid surfaces. As an example, we compute the lateral electrodynamic force between two corrugated surfaces over a wide parameter range. Universal behavior, fixed only by the largest wavelength component of the surface shape, is identified at large surface separations, complementing known short distance expansions which we also reproduce with high precision. PMID- 15903991 TI - Hyperbolic transverse patterns in nonlinear optical resonators. AB - We consider a nonlinear optical system in general, and a broad aperture laser, in particular, in a resonator where the diffraction coefficients are of opposite signs along two transverse directions. The system is described by the hyperbolic Maxwell-Bloch equations, where the spatial coupling is provided by the D'Alambert operator rather than by the Laplace operator. We show that this system supports hyperbolic transverse patterns residing on hyperbolas in far-field domain, and consisting of stretched vortices in near-field domain. PMID- 15903992 TI - Phase-space correlations of chaotic eigenstates. AB - It is shown that the Husimi representations of chaotic eigenstates are strongly correlated along classical trajectories. These correlations extend across the whole system size and, unlike the corresponding eigenfunction correlations in configuration space, they persist in the semiclassical limit. A quantitative theory is developed on the basis of Gaussian wave packet dynamics and random matrix arguments. The role of symmetries is discussed for the example of time reversal invariance. PMID- 15903993 TI - Random delays and the synchronization of chaotic maps. AB - We investigate the dynamics of an array of chaotic logistic maps coupled with random delay times. We report that for adequate coupling strength the array is able to synchronize, in spite of the random delays. Specifically, we find that the synchronized state is a homogeneous steady state, where the chaotic dynamics of the individual maps is suppressed. This synchronization behavior is largely independent of the connection topology and depends mainly on the average number of links per node. We carry out a statistical linear stability analysis that confirms the numerical results and provides a better understanding of the nontrivial roles of random delayed interactions. PMID- 15903994 TI - Chaotic mixing in a steady flow in a microchannel. AB - We report experiments on mixing of a passively advected fluorescent dye in a low Reynolds number flow in a microscopic channel. The channel is a chain of repeating segments with a custom designed profile that generates a steady three dimensional flow with stretching and folding, and chaotic mixing. A few statistical characteristics of mixing in the flow are studied and are all found to agree with theoretical and experimental results for the flows in the Batchelor regime of mixing that are chaotic in time. The proposed microchannel provides fast and efficient mixing and is simple to fabricate. PMID- 15903995 TI - Transition to spatiotemporal chaos in a two-dimensional hydrodynamic system. AB - We study the transition to spatiotemporal chaos in a two-dimensional hydrodynamic experiment where liquid columns take place in the gravity induced instability of a liquid film. The film is formed below a plane grid which is used as a porous media and is continuously supplied with a controlled flow rate. This system can be either ordered (on a hexagonal structure) or disordered depending on the flow rate. We observe, for the first time in an initially structured state, a subcritical transition to spatiotemporal disorder which arises through spatiotemporal intermittency. Statistics of numbers, creations, and fusions of columns are investigated. We exhibit a critical behavior close to the directed percolation one. PMID- 15903984 TI - Limit on the B0-->rho0rho0 branching fraction and implications for the Cabibbo Kobayashi-Maskawa angle alpha. AB - We search for the decay B0-->rho(0)rho(0) in a data sample of about 227x10(6) Upsilon(4S)-->BB decays collected with the BABAR detector at the PEP-II asymmetric-energy e(+)e(-) collider at SLAC. We find no significant signal and set an upper limit of 1.1x10(-6) at 90% C.L. on the branching fraction. As a result, the uncertainty due to penguin contributions on the Cabibbo-Kobayashi Maskawa unitarity angle alpha measured in B-->rhorho decays is decreased to 11 degrees at 68% C.L. PMID- 15903996 TI - Why the phase shifts for solitons on a vortex filament are so large: a theoretical explanation. AB - The phase "jumps" for solitons interacting on a vortex filament, observed in experiments, have been unaccounted for for more than 20 years. Using explicit formulas describing the interaction of two solitons on a thin vortex filament in the localized induction approximation, we show that an appropriate choice of the parameters of the solitons leads to large phase shifts. This result does not depend on the axial flow along the filament. PMID- 15903997 TI - Observation of the anomalous increase of the longitudinal energy spread in a space-charge-dominated electron beam. AB - We report a new experimental study of the growth of longitudinal energy spread in a space-charge-dominated electron beam, with a beam energy of several keV and beam current of approximately 100 mA. At relatively low beam densities, we measure growing energy spreads with distance along the transport channel, which are in remarkably good agreement with the theory of energy relaxation via Coulomb collisions. At higher beam densities, however, anomalous energy spreads exceeding the predictions of the relaxation theory are observed, which, we believe, could be caused by collective longitudinal-transverse instabilities observed in computer simulation studies. The onset of these instabilities occurs after several plasma periods according to calculations. PMID- 15903998 TI - Anomalous hollow electron beams in a storage ring. AB - This Letter reports the first observations of an anomalous hollow electron beam in the Duke storage ring. Created by exciting the single-bunch beam in a lattice with a negative chromaticity, the hollow beam consists of a solid core inside and a large ring outside. We report the detailed measurements of the hollow beam phenomenon, including its distinct image pattern, spectrum signature, and its evolution with time. By capturing the postinstability bursting beam, the hollow beam is a unique model system for studying the transverse instabilities, in particular, the interplay of the wakefield and the lattice nonlinearity. In addition, the hollow beam can be used as a powerful tool to study the linear and nonlinear particle dynamics in the storage ring. PMID- 15903999 TI - Self-regulation of E x B flow shear via plasma turbulence. AB - The momentum balance has been applied to the ExB flow in the edge region of a reversed field pinch (RFP) configuration. All terms, including those involving fluctuations, have been measured in stationary condition in the edge region of the Extrap-T2R RFP experiment. It is found that the component of the Reynolds stress driven by electrostatic fluctuations is the term playing the major role in driving the shear of the ExB flow to a value marginal for turbulent suppression, so that the results are in favor of a turbulence self-regulating mechanism underlying the momentum balance at the edge. Balancing the sheared flow driving and damping terms, the plasma viscosity is found anomalous and consistent with the diffusivity due to electrostatic turbulence. PMID- 15904000 TI - Measurement of turbulence decorrelation during transport barrier evolution in a high-temperature fusion plasma. AB - A low power polychromatic beam of microwaves is used to diagnose the behavior of turbulent fluctuations in the core of the JT-60U tokamak during the evolution of the internal transport barrier. A continuous reduction in the size of turbulent structures is observed concomitant with the reduction of the density scale length during the evolution of the internal transport barrier. The density correlation length decreases to the order of the ion gyroradius, in contrast with the much longer scale lengths observed earlier in the discharge, while the density fluctuation level remain similar to the level before transport barrier formation. PMID- 15904001 TI - Cross-scale nonlinear coupling and plasma energization by Alfven waves. AB - We present a new channel for the nonlocal transport of wave energy from the large (MHD) scales to the small (kinetic) scales generated by the resonant decay of MHD Alfven waves into kinetic Alfven waves. This process does not impose any restriction on the wave numbers or frequencies of initial MHD waves, which makes it superior compared to the mechanisms of spectral transport studied before. Because of dissipative properties of the nonlinearly driven kinetic Alfven waves, the decay leads to plasma heating and particle acceleration, which is observed in a variety of space and astrophysical plasmas. Two examples in the solar corona and the terrestrial magnetosphere are briefly discussed. PMID- 15904002 TI - The CERN Large Hadron Collider as a tool to study high-energy density matter. AB - The Large Hadron Collider (LHC) at CERN will generate two extremely powerful 7 TeV proton beams. Each beam will consist of 2808 bunches with an intensity per bunch of 1.15x10(11) protons so that the total number of protons in one beam will be about 3x10(14) and the total energy will be 362 MJ. Each bunch will have a duration of 0.5 ns and two successive bunches will be separated by 25 ns, while the power distribution in the radial direction will be Gaussian with a standard deviation, sigma=0.2 mm. The total duration of the beam will be about 89 mus. Using a 2D hydrodynamic code, we have carried out numerical simulations of the thermodynamic and hydrodynamic response of a solid copper target that is irradiated with one of the LHC beams. These calculations show that only the first few hundred proton bunches will deposit a high specific energy of 400 kJ/g that will induce exotic states of high energy density in matter. PMID- 15904003 TI - Hydrogen-bond topology and the ice VII/VIII and ice Ih/XI proton-ordering phase transitions. AB - The existence of an ice Ih/XI proton-ordering transition to a low-temperature ferroelectric phase has sparked considerable debate in the literature. Electronic density functional theory calculations, extended using graph invariants, confirm that a transition to a low-temperature ferroelectric phase should occur. The predicted transition at 98 K is in qualitative agreement with the observed transition at 72 K, and the low-temperature phase is the ferroelectric phase determined in diffraction experiments. The theoretical methods used to predict the phase transition are validated by comparing their prediction to the well characterized ice VII/VIII proton-ordering transition. PMID- 15904004 TI - Critical Casimir effect in three-dimensional Ising systems: measurements on binary wetting films. AB - The critical Casimir force (CF) is observed in thin wetting films of a binary liquid mixture close to the liquid/vapor coexistence. X-ray reflectivity shows thickness (L) enhancement near the bulk consolute point. The extracted Casimir amplitude Delta(+-)=3+/-1 agrees with the theoretical universal value for the antisymmetric 3D Ising films. The onset of CF in the one-phase region occurs at L/xi approximately 5 regardless of whether the bulk correlation length xi is varied with temperature or composition. The shape of the Casimir scaling function depends monotonically on the dimensionality. PMID- 15904005 TI - Structural arrest in an ideal gas. AB - We report a molecular dynamics study of a simple model system that has the static properties of an ideal gas, yet exhibits nontrivial "glassy" dynamics behavior at high densities. The constituent molecules of this system are constructs of three infinitely thin hard rods of length L, rigidly joined at their midpoints. The crosses have random but fixed orientation. The static properties of this system are those of an ideal gas, and its collision frequency can be computed analytically. For number densities NL(3)/V>>1, the single-particle diffusivity goes to zero. As the system is completely structureless, standard mode-coupling theory cannot describe the observed structural arrest. Nevertheless, the system exhibits many dynamical features that appear to be mode-coupling-like. All high density incoherent intermediate scattering functions collapse onto master curves that depend only on the wave vector. PMID- 15904006 TI - Self-organized growth of nanopucks on Pb quantum islands. AB - Electronic Moire patterns found on lead (Pb) quantum islands can serve as a template to grow self-organized cluster (nanopucks) arrays of various materials. These patterns can be divided into fcc- and hcp-stacked areas, which exhibit different binding strengths to the deposited adatoms. For Ag adatoms, the binding energy can differ substantially and the confined nucleation thus occurs in the fcc sites. Both the size distribution and spatial arrangement of the Ag nanopucks are analyzed and found to be commensurate with the characteristics of the template island, which exhibits a bilayer oscillatory behavior. PMID- 15904007 TI - Two-dimensional metal-insulator transition as a percolation transition in a high mobility electron system. AB - By carefully analyzing the low temperature density dependence of 2D conductivity in undoped high-mobility n-GaAs heterostructures, we conclude that the 2D metal insulator transition in this 2D electron system is a density inhomogeneity driven percolation transition due to the breakdown of screening in the random charged impurity disorder background. In particular, our measured conductivity exponent of approximately 1.4 approaches the 2D percolation exponent value of 4/3 at low temperatures and our experimental data are inconsistent with there being a zero temperature quantum critical point in our system. PMID- 15904008 TI - Electronic origin of solid solution softening in bcc molybdenum alloys. AB - The intrinsic mechanism of solid solution softening in bcc molybdenum alloys due to 5d transition metal additions is investigated on the basis of ab initio electronic-structure calculations that model the effect of alloying elements on the generalized stacking fault (GSF) energies. We demonstrate that additions with an excess of electrons (Re, Os, Ir, and Pt) lead to a decrease in the GSF energy and those with a lack of electrons (Hf and Ta) to its sharp increase. Using the generalized Peierls-Nabarro model for a nonplanar core, we associate the local reduction of the GSF energy with an enhancement of double kink nucleation and an increase of the dislocation mobility, and we reveal the electronic reasons for the observed dependence of the solution softening on the atomic number of the addition. PMID- 15904009 TI - Cascade of bulk magnetic phase transitions in NaxCoO2 as studied by muon spin rotation. AB - Using muon spin rotation, well-defined bulk approximately 100% magnetic phases in NaxCoO2 are revealed. A novel magnetic phase is detected for x=0.85 with the highest transition temperature ever observed for x>or=0.75. This stresses the diversity of x>or=0.75 magnetic phases and the link between magnetic and structural degrees of freedom. For the charge-ordered x=0.50 compound, a cascade of transitions is observed below 85 K. From a detailed analysis of our data, we conclude that the ordered moment varies continuously with temperature and suggest that the two secondary transitions at 48 and 29 K correspond to a moderate reorientation of antiferromagnetically coupled moments. PMID- 15904010 TI - Determination of a merocyanine J-aggregate structure and the significant contribution of the electric dipole interaction to the exciton band wavelength. AB - The molecular alignment of a merocyanine (MC) J-aggregate monolayer at the air water interface was determined by a grazing incidence x-ray diffraction method. The obtained molecular arrangement apparently shows that the conventional formula, which accounts only for the transition dipole interaction, is not sufficient to figure out the exciton band wavelength, suggesting the importance of the electric dipole (ED) interaction. We derived a simple formula for the ED interaction energy under an extended dipole approximation and clarified the ED contribution in the MC J aggregate. PMID- 15904011 TI - Junction of three quantum wires: restoring time-reversal symmetry by interaction. AB - We investigate the transport of correlated fermions through a junction of three one-dimensional quantum wires pierced by a magnetic flux. We determine the flow of the conductance as a function of a low-energy cutoff in the entire parameter space. For attractive interactions and generic flux the fixed point with maximal asymmetry of the conductance is the stable one, as conjectured recently. For repulsive interactions and arbitrary flux we find a line of stable fixed points with vanishing conductance as well as stable fixed points with symmetric conductance (4/9)(e(2)/h). PMID- 15904012 TI - Signature of Mott-insulator transition with ultracold fermions in a one dimensional optical lattice. AB - Using the exact Bethe ansatz solution of the Hubbard model and Luttinger liquid theory, we investigate the density profiles and collective modes of one dimensional ultracold fermions confined in an optical lattice with a harmonic trapping potential. We determine a generic phase diagram in terms of a characteristic filling factor and a dimensionless coupling constant. The collective oscillations of the atomic mass density, a technique that is commonly used in experiments, provide a signature of the quantum phase transition from the metallic phase to the Mott-insulator phase. A detailed experimental implementation is proposed. PMID- 15904013 TI - Coherence of elementary excitations in a disordered electron system. AB - The localization properties of the single-particle and collective electron excitations were investigated in the intentionally disordered GaAs/AlGaAs superlattices by weak-field magnetoresistance and Raman scattering. The localization length of the individual electron was found to be considerably larger than that of the collective excitations. This suggests that the disorder has a weaker effect on the electrons than on their collective motion and that the interaction which gives rise to the collective effects increases localization. PMID- 15904014 TI - Singular effect of disorder on electronic transport in strongly coupled electron phonon systems. AB - We solve the disordered Holstein model in three dimensions considering the phonon variables to be classical. After mapping out the phases of the "clean" strong coupling problem, we focus on the effect of disorder at strong electron-phonon (EP) coupling. The presence of even weak disorder (i) enormously enhances the resistivity (rho) at T=0, simultaneously suppressing the density of states at the Fermi level, (ii) suppresses the temperature dependent increase of rho, and (iii) leads to a regime with drho/dT<0. We locate the origin of these anomalies in the disorder induced tendency towards polaron formation, and the associated suppression in effective carrier density and mobility. These results, explicitly at "metallic" density, are of direct relevance to disordered EP materials such as covalent semiconductors, the manganites, and to anomalous transport in the A-15 compounds. PMID- 15904015 TI - Deformation electron-phonon coupling in disordered semiconductors and nanostructures. AB - We study the electron-phonon relaxation (dephasing) rate in disordered semiconductors and low-dimensional structures. The relaxation is determined by the interference of electron scattering via the deformation potential and elastic electron scattering from impurities and defects. We have found that in contrast with the destructive interference in metals, which results in the Pippard ineffectiveness condition for the electron-phonon interaction, the interference in semiconducting structures substantially enhances the effective electron-phonon coupling. The obtained results provide an explanation to energy relaxation in silicon structures. PMID- 15904016 TI - Weak Coulomb blockade effect in quantum dots. AB - We develop the general nonequilibrium theory of transport through a quantum dot, including Coulomb blockade effects via a 1/N expansion, where N is the number of scattering channels. At lowest order we recover the Landauer formula for the current plus a self-consistent equation for the dot potential. We obtain the leading corrections and compare with earlier approaches. Finally, we show that to leading and to next leading order in 1/N there is no interaction correction to the weak localization, in contrast to previous theories, but consistent with experiments by Huibers et al. [Phys. Rev. Lett. 81, 1917 (1998)], where N=4. PMID- 15904017 TI - Spatially dependent inelastic tunneling in a single metallofullerene. AB - We have measured the elastic and inelastic tunneling properties of isolated Gd@C(82) molecules on Ag(001) using cryogenic scanning tunneling spectroscopy. We find that the dominant inelastic channel is spatially well localized to a particular region of the molecule. Ab initio pseudopotential density-functional theory calculations indicate that this channel arises from a vibrational cage mode. We further show that the observed inelastic tunneling localization is explained by strong localization in the molecular electron-phonon coupling to this mode. PMID- 15904018 TI - Fractional-flux vortices and spin superfluidity in triplet superconductors. AB - We discuss a novel type of fractional-flux vortices along with integer flux vortices in Kosterlitz-Thouless transitions in a triplet superconductor. We show that under certain conditions a spin-triplet superconductor should exhibit a novel state of spin superfluidity without superconductivity. PMID- 15904019 TI - Helical vortex phase in the noncentrosymmetric CePt3Si. AB - We consider the role of magnetic fields on the broken inversion superconductor CePt3Si. We show that the upper critical field for a field along the c axis exhibits a much weaker paramagnetic effect than for a field applied perpendicular to the c axis. The in-plane paramagnetic effect is strongly reduced by the appearance of helical structure in the order parameter. We find that, to get good agreement between theory and recent experimental measurements of H(c2), this helical structure is required. We propose a Josephson junction experiment that can be used to detect this helical order. In particular, we predict that the Josephson current will exhibit a magnetic interference pattern for a magnetic field applied perpendicular to the junction normal. We also discuss unusual magnetic effects associated with the helical order. PMID- 15904020 TI - Effect of antiferromagnetic planes on the superconducting properties of multilayered high-Tc cuprates. AB - We propose a mechanism for high critical temperature (T(c)) in the coexistent phase of superconducting (SC) and antiferromagnetic (AFM) CuO2 planes in multilayered cuprates. The Josephson coupling between the SC planes separated by an AFM insulator (Mott insulator) is calculated perturbatively up to the fourth order in terms of the hopping integral between adjacent CuO2 planes. It is shown that the AFM exchange splitting in the AFM plane suppresses the so-called pi Josephson coupling, and the long-ranged 0-Josephson coupling leads to coexistence with a rather high value of T(c). PMID- 15904021 TI - Coherence and superconductivity in coupled one-dimensional chains: a case study of YBa2Cu3Oy. AB - We report the infrared (IR) response of Cu-O chains in the high-T(c) superconductor YBa(2)Cu(3)O(y) over the doping range spanning y=6.28-6.75. We find evidence for a power law scaling at mid-IR frequencies consistent with predictions for Tomonaga-Luttinger liquid, thus supporting the notion of one dimensional transport in the chains. We analyze the role of coupling to the CuO2 planes in establishing metallicity and superconductivity in disordered chain fragments. PMID- 15904022 TI - Electric polarization rotation in a hexaferrite with long-wavelength magnetic structures. AB - We report on the control of electric polarization (P) by using magnetic fields (B) in a hexaferrite having magnetic order above room temperature (RT). The material investigated is hexagonal Ba0.5Sr1.5Zn2Fe12O22, which is a nonferroelectric helimagnetic insulator in the zero-field ground state. By applying B, the system undergoes successive metamagnetic transitions, and shows concomitant ferroelectric order in some of the B-induced phases with long wavelength magnetic structures. The magnetoelectrically induced P can be rotated 360 degrees by external B. This opens up the potential for not only RT magnetoelectric devices but also devices based on the magnetically controlled electro-optical response. PMID- 15904023 TI - Probing spin correlations with phonons in the strongly frustrated magnet ZnCr2O4. AB - The spin-lattice coupling plays an important role in strongly frustrated magnets. In ZnCr2O4, an excellent realization of the Heisenberg antiferromagnet on the pyrochlore network, a lattice distortion relieves the geometrical frustration through a spin-Peierls-like phase transition at T(c)=12.5 K. Conversely, spin correlations strongly influence the elastic properties of a frustrated magnet. By using infrared spectroscopy and published data on magnetic specific heat, we demonstrate that the frequency of an optical phonon triplet in ZnCr2O4 tracks the nearest-neighbor spin correlations above T(c). The splitting of the phonon triplet below T(c) provides a way to measure the spin-Peierls order parameter. PMID- 15904024 TI - Ground state of magnetic dimers on metal surfaces. AB - We present model studies of the ground state for magnetic dimers on metal surfaces. We find it can be neither ferromagnetic nor antiferromagnetic, but is often canted for nearest neighbors. Thus, the system cannot be described using bilinear exchange. We give a criterion which can be used quite generally to interrogate the local stability of ferromagnetically or antiferromagnetically aligned dimers, and which also may be used to infer the canting angle when canted states are stable. PMID- 15904025 TI - Magnetization reversal in spin patterns with complex geometry. AB - We study field-driven dynamics of spins with antiferromagnetic interactions along the links of a complex substrate geometry, which is modeled by graphs of a controlled connectivity distribution. The magnetization reversal occurs in avalanches of spin flips, which are pinned by the topological constraints of the underlying graph. The hysteresis loop and avalanche sizes are analyzed and classified in terms of the graph's connectivity and clustering. The results are relevant for magnets with a hierarchical spatial inhomogeneity and for design of nanoscale magnetic devices. PMID- 15904026 TI - Helicoidal ordering in iron perovskites. AB - We study the double exchange in transition metal oxides with itinerant and localized electrons. We show that the charge transfer energy Delta and the oxygen oxygen hopping amplitude t(pp) have a strong effect on magnetic ordering: while for Delta>0 the ground state is ferromagnetic, for negative Delta and large t(pp) the double exchange gives rise to an incommensurate helicoidal ordering of local spins, observed, e.g., in the iron perovskites SrFeO3 and CaFeO3. For negative Delta, the metal-insulator transition into a charge-ordered state has little effect on magnetic ordering. This explains the difference in magnetic and transport properties of ferrates and manganites. PMID- 15904027 TI - Carrier-induced magnetic ordering control in a digital (Ga,Mn)as structure. AB - With the full potential linearized augmented plane method, we theoretically investigated the carrier-induced magnetization reversal in digital (Ga,Mn)As heterostructures with varying distance between the two Mn layers along with the distribution and concentration of external carriers. The presence of external holes induces switching from the antiferromagnetic to ferromagnetic state when d(Mn-Mn)=16.96 A, whereas the addition of electrons produces no significant effect. We demonstrate a possibility to separately control T(c) and magnetic reversal in digital (Ga,MN)As alloys. PMID- 15904028 TI - Competing effects of interactions and spin-orbit coupling in a quantum wire. AB - We study the interplay of electron-electron interactions and Rashba spin-orbit coupling in one-dimensional ballistic wires. Using the renormalization group approach we construct the phase diagram in terms of Rashba coupling, Tomonaga Luttinger stiffness and backward scattering strength. We identify the parameter regimes with a dynamically generated spin gap and show where the Luttinger liquid prevails. We also discuss the consequences for the operation of the Datta-Das transistor. PMID- 15904029 TI - Spin waves in nickel nanorings of large aspect ratio. AB - The spin dynamics of high-aspect-ratio nickel nanorings in a longitudinal magnetic field have been investigated by Brillouin spectroscopy and the results are compared with a macroscopic theory and three-dimensional micromagnetic simulations. Good agreement is found between the measured and calculated magnetic field dependence of the spin wave frequency. Simulations show that as the field decreases from saturation, the rings switch from a "bamboo" to a novel "twisted bamboo" state at a certain critical field, and predict a corresponding dip in the dependence of the spin wave frequency on the magnetic field. PMID- 15904030 TI - NMR evidence for triple-q multipole structure in NpO2. AB - In order to elucidate the nature of the exotic ordered phase of NpO2 below T(0)=26 K, we have initiated the first 17O-NMR measurements on this system. From the 17O-NMR spectrum, the occurrence of two inequivalent oxygen sites has been confirmed below T0. It has also been shown that the characteristic features of the hyperfine interaction at the oxygen sites are well explained by invoking a hyperfine interaction with field-induced antiferromagnetic moments which appear as a result of the triple-q antiferroquadrupolar order. The NMR findings strongly support the occurrence of the longitudinal triple-q multipole structure in NpO2. PMID- 15904031 TI - Body-centered-cubic Ni and its magnetic properties. AB - The body-centered-cubic (bcc) phase of Ni, which does not exist in nature, has been achieved as a thin film on GaAs(001) at 170 K via molecular beam epitaxy. The bcc Ni is ferromagnetic with a Curie temperature of 456 K and possesses a magnetic moment of 0.52+/-0.08 micro(B)/atom. The cubic magnetocrystalline anisotropy of bcc Ni is determined to be +4.0x10(5) ergs x cm(-3), as opposed to 5.7x10(4) ergs x cm(-3) for the naturally occurring face-centered-cubic (fcc) Ni. This sharp contrast in the magnetic anisotropy is attributed to the different electronic band structures between bcc Ni and fcc Ni, which are determined using angle-resolved photoemission with synchrotron radiation. PMID- 15904032 TI - Polarization of valence band holes in the (Ga,Mn)as diluted magnetic semiconductor. AB - We report on direct measurements of the impurity band hole polarization in the diluted magnetic semiconductor (Ga,Mn)As. The polarization of impurity band holes in a magnetic field is strongly enhanced by antiferromagnetic exchange interaction with Mn ions. The temperature dependence of the hole polarization shows a strong increase of this polarization below the Curie temperature. We show that the ground state of the impurity band is formed by uniaxial stress split F=+/-1 states of antiferromagnetically coupled Mn ions (S=5/2) and valence band holes (J=3/2). The gap between the Mn acceptor related impurity band and the valence band is directly measured in a wide range of Mn content. PMID- 15904033 TI - Ultrafast breathinglike oscillation in the exciton density of ZnSe quantum wells. AB - The spatial density profile of a low-density exciton ensemble in ZnSe quantum wells shows a breathinglike oscillation on a 30-ps time scale. This breathing results from the emission of the first acoustic phonon at the end of the quasiballistic transport phase of the excitons which reverses their direction of propagation. Since the scattering destroys the phase of the excitonic wave function, one can deduce simultaneously the coherence length and the coherence time of excitonic transport by evaluation of the oscillation measured from a single experiment. The breathing, which can be modeled by Monte Carlo simulations, is quenched for rising lattice temperature, i.e., increasing phonon absorption, and in samples with significant disorder. These results were obtained by time-resolved nanophotoluminescence with 5 ps and 250 nm temporal and spatial resolution, respectively. PMID- 15904034 TI - Yield of singlet excitons in organic light-emitting devices: a double modulation photoluminescence-detected magnetic resonance study. AB - Double-modulation (DM) photoluminescence (PL) detected magnetic resonance (PLDMR) measurements on poly(2-methoxy-5-(2(')-ethyl)-hexoxy-1,4-phenylene vinylene) are described. In these measurements, the laser excitation power is modulated at 10(+) pure Fermi decay of (38)K(m) is a =0.9981+/-0.0030+0.0032 / -0.0037, consistent with the standard model prediction a =1. PMID- 15904060 TI - Resonant activation in a nonadiabatically driven optical lattice. AB - We demonstrate the phenomenon of resonant activation in a nonadiabatically driven dissipative optical lattice with broken time symmetry. The resonant activation results in a resonance as a function of the driving frequency in the current of atoms through the periodic potential. We demonstrate that the resonance is produced by the interplay between deterministic driving and fluctuations, and we also show that by changing the frequency of the driving it is possible to control the direction of the diffusion. PMID- 15904061 TI - Field-free three dimensional molecular axis alignment. AB - We investigate strategies for field-free three dimensional molecular axis alignment using strong nonresonant laser fields under experimentally realistic conditions. Using the polarizabilites and rotational constants of an asymmetric top rotor molecule (ethene, C2H4), we consider three different methods for axis alignment of a Boltzmann distribution of rotors at 4 K. Specifically, we compare the use of impulsive kick laser pulses having both linear and elliptical polarization to the use of elliptically polarized switched laser pulses. We show that an enhanced degree of field-free three dimensional alignment of ground vibronic state molecules obtains from the use of two orthogonally polarized, time separated laser pulses. PMID- 15904062 TI - Exact time-dependent exchange-correlation potentials for strong-field electron dynamics. AB - By solving the time-dependent Schrodinger equation and inverting the time dependent Kohn-Sham scheme we obtain the exact time-dependent exchange correlation potential of density-functional theory for the strong-field dynamics of a correlated system. We demonstrate that essential features of the exact exchange-correlation potential can be related to derivative discontinuities in stationary density-functional theory. Incorporating the discontinuity in a time dependent density-functional calculation greatly improves the description of the ionization process. PMID- 15904063 TI - Accumulation of three-body resonances above two-body thresholds. AB - We calculate resonances in the three-body system with attractive Coulomb potential by solving homogeneous Faddeev-Merkuriev integral equations for complex energies. The equations are solved using the Coulomb-Sturmian separable expansion method. This approach allows us to study the exact behavior of the three-body Coulomb systems near the threshold. A negatively charged positronium ion is used as a test case. In addition to locating all previously known S-wave resonances of the positronium ion, we also find a large number of new resonant states that accumulate just slightly above the two-body thresholds. The pattern of accumulation of resonant states above the two-body thresholds suggests that probably they are infinite in number. We conjecture that this may be a general property of the three-body system with an attractive Coulomb potential. PMID- 15904064 TI - Dynamics of traveling waves in the transverse section of a laser. AB - We analyze the general features of the formation and interaction of transverse traveling waves and the appearance of filamentation in broad area semiconductor lasers with current profiling. For small apertures, the emitted profile is symmetric consisting of two counterpropagating transverse traveling waves, both emanating from the center of the device. For larger apertures, the emission becomes asymmetric as one of the traveling waves expands to occupy an increased area while the other occupies the remaining, smaller spatial region. In both devices, the pattern becomes unstable at higher injection currents due to optical filamentation, although an intermediate state is present in the wider device whereby the dominant wave undergoes a Hopf bifurcation before filamentation occurs. PMID- 15904065 TI - Complete modal decomposition for optical waveguides. AB - Virtually all electromagnetic waveguiding structures support a multiplicity of modes. Nevertheless, to date, an experimental method for unique decomposition of the fields in terms of the component eigenmodes has not been realized. The fundamental problem is that all current attempts of modal decomposition do not yield phase information. Here we introduce a noninterferometric approach to achieve modal decomposition of the fields at the output of a general waveguiding structure. The technique utilizes a mapping of the two-dimensional field distribution onto the one-dimensional space of waveguide eigenmodes, together with a phase-retrieval algorithm to extract the amplitudes and phases of all the guided vectorial modes. Experimental validation is provided by using this approach to examine the interactions of 16 modes in a hollow-core photonic-band gap fiber. PMID- 15904066 TI - Nanoscale resolution in the focal plane of an optical microscope. AB - Utilizing single fluorescent molecules as probes, we prove the ability of a far field microscope to attain spatial resolution down to 16 nm in the focal plane, corresponding to about 1/50 of the employed wavelength. The optical bandwidth expansion by nearly an order of magnitude is realized by a saturated depletion through stimulated emission of the molecular fluorescent state. We demonstrate that en route to the molecular scale, the resolving power increases with the square root of the saturation level, which constitutes a new law regarding the resolution of an emerging class of far-field light microscopes that are not limited by diffraction. PMID- 15904067 TI - Direct processes in chaotic microwave cavities in the presence of absorption. AB - We quantify the presence of direct processes in the S matrix of chaotic microwave cavities with absorption in the one-channel case. To this end the full distribution P(S)(S) of the S matrix, i.e., S=sqrt[R]e(itheta), is studied in cavities with time-reversal symmetry for different antenna coupling strengths T(a) or direct processes. The experimental results are compared with random matrix calculations and with numerical simulations including absorption. The theoretical result is a generalization of the Poisson kernel. The experimental and the numerical distributions are in excellent agreement with theoretical predictions for all cases. PMID- 15904057 TI - Measurement of branching fractions and charge asymmetries for exclusive B decays to charmonium. AB - We report measurements of branching fractions and charge asymmetries of exclusive decays of neutral and charged B mesons into two-body final states containing a charmonium state and a light strange meson. The charmonium mesons considered are J/psi, psi(2S) and chi(c1), and the light meson is either K or K(*). We use a sample of about 124x10(6) BB pairs collected with the BABAR detector at the PEP II storage ring at the Stanford Linear Accelerator Center. PMID- 15904068 TI - Magnetically controlled convection in a diamagnetic fluid. AB - We present visualization and measurement of the convection of water under a high magnetic field applied vertically to the fluid. The convection was either suppressed or enhanced depending on the direction of the magnetic force. The magnetic field effect was evaluated quantitatively by measuring the onset of convection, and discussed in terms of the Rayleigh number which includes the magnetic term. The results clearly show that the convection in a diamagnetic fluid such as water can be controlled using a common 10 T class magnet. PMID- 15904070 TI - Dominant electrostatic nature of the reversed field pinch dynamo. AB - The origin of the dynamo velocity field of the reversed field pinch within the visco-resistive MHD modeling is uncovered. The main component of this field is an electrostatic drift. The corresponding electrostatic field is related to a small charge separation which is consistent with the quasineutrality approximation, and which should be present in real plasmas, too. While quite natural in the stationary single helicity state, this analysis is shown to extend also to the nonstationary multiple helicity regime. Numerical simulations provide the spatial distribution of fields and of charge separation. PMID- 15904071 TI - Global gyrokinetic stability of pressure-gradient-driven electromagnetic modes in tokamaks with regions of low shear. AB - Tokamaks with large pressure gradients (alpha(max)) formed in regions of weak magnetic shear are shown to be susceptible to novel, low-n, global, kinetic, electromagnetic modes with a toroidal mode number n in the range 21 on the magnetic axis and alpha(max) near q(min), new, global kinetic infernal modes with a strong mode rotation omega(r) and a finite growth rate gamma<|omega(r)| are found. For equilibria with reverse shear where q(min) is off axis and alpha(max) near q(min), the existence of an unstable low-n global branch of Alfve n ion temperature gradient modes is revealed with an oscillatory gamma as a function of n. The addition of trapped electron dynamics is shown to be further destabilizing. PMID- 15904069 TI - Observation of the acceleration of a single bunch by using the induction device in the KEK proton synchrotron. AB - A single rf bunch in the KEK proton synchrotron was accelerated with an induction acceleration method from the injection energy of 500 MeV to 5 GeV. PMID- 15904072 TI - Electronic-enthalpy functional for finite systems under pressure. AB - We introduce the notion of electronic enthalpy for first-principles structural and dynamical calculations of finite systems under pressure. An external pressure field is allowed to act directly on the electronic structure of the system studied via the ground-state minimization of the functional E+PV(q), where V(q) is the quantum volume enclosed by a charge isosurface. The Hellmann-Feynman theorem applies, and assures that the ionic equations of motion follow an isoenthalpic dynamics. No pressurizing medium is explicitly required, while coatings of environmental ions or ligands can be introduced if chemically relevant. We apply this novel approach to the study of group-IV nanoparticles during a shock wave, highlighting the significant differences in the plastic or elastic response of the diamond cage under load, and their potential use as novel nanostructured impact-absorbing materials. PMID- 15904073 TI - Slow energy relaxation of macromolecules and nanoclusters in solution. AB - Many systems in the realm of nanophysics from both the living and the inorganic world display slow relaxation kinetics of energy fluctuations. In this Letter we propose a general explanation for such a phenomenon, based on the effects of interactions with the solvent. Within a simple harmonic model of the system fluctuations, we demonstrate that the inhomogeneity of coupling to the solvent of the bulk and surface atoms suffices to generate a complex spectrum of decay rates. We show for myoglobin and for a metal nanocluster that the result is a complex, nonexponential relaxation dynamics. PMID- 15904074 TI - Strong dc electric field applied to supersaturated aqueous glycine solution induces nucleation of the gamma polymorph. AB - Applying a strong static electric field to supersaturated aqueous glycine solutions resulted in the nucleation of the gamma polymorph. This is the first report of a strong dc field inducing the nucleation of a neutral solute in a supersaturated solution. We attribute this effect to the electric-field-induced orientation of the highly polar glycine molecules in large preexisting solute clusters, helping them organize into a crystalline structure. This result also lends further support to our proposed optical-Kerr mechanism for nonphotochemical laser-induced nucleation. PMID- 15904075 TI - Real-time evolution of the distribution of nanoparticles in an ultrathin-polymer film-based waveguide. AB - We report a novel application of an ultrathin-polymer-film-based, resonance enhanced x-ray waveguide as a real-time nanoprobe for elucidating dilute, yet disordered, gold nanoparticles embedded in the polymer matrix. This nanoprobe promises a sensitivity enhancement of several orders of magnitude, hence revealing in real time the lateral nanoparticle distribution with subnanometer spatial resolution. We observed that the motion of the nanoparticles is strongly anisotropic, with in-plane coalescence taking place more rapidly than out-of plane diffusion, which can ultimately facilitate the formation of two-dimensional structures. PMID- 15904076 TI - Atomistic deformation modes in strong covalent solids. AB - We report on a first-principles study of the structural deformation modes in diamond, cubic boron nitride (c-BN), and cubic BC2N. We show that (i) the diamond C-C bonds remain strong up to the breaking point, leading to the large and nearly identical shear and tensile strength, (ii) c-BN exhibits a shear failure mode different from that in diamond and a significant softening in the B-N bonds at large tensile strains long before the bond breaking, and (iii) cubic BC2N displays a large disparity between the shear and tensile strength, contrary to the expectation for the hybrid of diamond and c-BN. We examine the microscopic bond-breaking processes to elucidate the atomistic mechanisms for the deformation modes and the implications for material strength. PMID- 15904077 TI - Modeling the phase diagram of carbon. AB - We determined the phase diagram involving diamond, graphite, and liquid carbon using a recently developed semiempirical potential. Using accurate free-energy calculations, we computed the solid-solid and solid-liquid phase boundaries for pressures and temperatures up to 400 GPa and 12 000 K, respectively. The graphite diamond transition line that we computed is in good agreement with experimental data, confirming the accuracy of the employed empirical potential. On the basis of the computed slope of the graphite melting line, we rule out the hotly debated liquid-liquid phase transition of carbon. Our simulations allow us to give accurate estimates of the location of the diamond melting curve and of the graphite-diamond-liquid triple point. PMID- 15904078 TI - Asymmetrically coupled directed percolation systems. AB - We introduce a dynamical model of coupled directed percolation systems with two particle species. The two species A and B are coupled asymmetrically in that A particles branch B particles, whereas B particles prey on A particles. This model may describe epidemic spreading controlled by reactive immunization agents. We study nonequilibrium phase transitions with attention focused on the multicritical point where both species undergo the absorbing phase transition simultaneously. In one dimension, we find that the inhibitory coupling from B to A is irrelevant and the model belongs to the unidirectionally coupled directed percolation class. On the contrary, a mean-field analysis predicts that the inhibitory coupling is relevant and a new universality appears with a variable dynamic exponent. Numerical simulations on small-world networks confirm our predictions. PMID- 15904079 TI - Compressive stress in polycrystalline volmer-weber films. AB - The Volmer-Weber mode for growing polycrystalline films, which comprises island, network, and channel stages before the films become continuous, is well known for its complex stress behavior with compressive and tensile stress alternating in the initial three growth stages. Recently, two new mechanisms for the compressive stress have been proposed [Phys. Rev. Lett. 88, 156103 (2002); 89, 126103 (2002)], which account for the reversibility of stress generation and relaxation. We show that the two mechanisms play only minor roles for the development of compressive stress, which is confirmed to be due to capillarity effects in the precoalescence stage. PMID- 15904080 TI - Columnar AlGaN/GaN nanocavities with AlN/GaN Bragg reflectors grown by molecular beam epitaxy on Si(111). AB - Self-assembled columnar AlGaN/GaN nanocavities, with an active region of GaN quantum disks embedded in an AlGaN nanocolumn and cladded by top and bottom AlN/GaN Bragg mirrors, were grown. The nanocavity has no cracks or extended defects, due to the relaxation at the Si interface and to the nanocolumn free surface to volume ratio. The emission from the active region matched the peak reflectivity by tuning the Al content and the GaN disks thickness. Quantum confinement effects that depend on both the disk thickness and the inhomogeneous strain distribution within the disks are clearly observed. PMID- 15904081 TI - Patterning multilayers of molecules via self-organization. AB - The electric dipole interaction among adsorbate molecules may cause them to form regular nanopatterns. In a multilayer system, the self-organization of each layer is also influenced by the underlying layers. This Letter develops a phase field model to simulate the molecular patterning process. The study reveals self alignment, scaling down of size, and the effect of guided self-assembly with embedded electrodes. PMID- 15904082 TI - Bond breaking coupled with translation in rolling of covalently bound molecules. AB - The response of a C60 molecule to manipulation across a surface displays a long range periodicity which corresponds to a rolling motion. A period of three or four lattice constants is observed and is accompanied by complex subharmonic structure due to molecular hops through a regular, repeating sequence of adsorption states. Combining experimental data and ab initio calculations, we show that this response corresponds to a rolling motion in which two of the four Si-C60 covalent bonds act as a pivot over which the molecule rotates while moving through one lattice constant and identify a sequence of C60 bonding configurations that accounts for the periodic structure. PMID- 15904083 TI - Strain relief in Cu-Pd heteroepitaxy. AB - We present experimental and theoretical studies of Pd/Cu(100) and Cu/Pd(100) heterostructures in order to explore their structure and misfit strain relaxation. Ultrathin Pd and Cu films are grown by pulsed laser deposition at room temperature. For Pd/Cu, compressive strain is released by networks of misfit dislocations running in the [100] and [010] directions, which appear after a few monolayers (ML) already. In striking contrast, for Cu/Pd the tensile overlayer remains coherent up to about 9 ML, after which multilayer growth occurs. The strong asymmetry between tensile and compressive cases is in contradiction with continuum elasticity theory and is also evident in the structural parameters of the strained films. Molecular dynamics calculations based on classical many-body potentials confirm the pronounced tensile-compressive asymmetry and are in good agreement with the experimental data. PMID- 15904084 TI - Ab initio design of high-k dielectrics: La(x)Y1-xAlO3. AB - We use calculations based on density-functional theory in the virtual crystal approximation for the design of high-k dielectrics, which could offer an alternative to silicon dioxide in complementary metal-oxide semiconductor devices. We show that aluminates LaxY1-xAlO3 alloys derived by mixing aluminum oxide with lanthanum and yttrium oxides have unique physical attributes for a possible application as gate dielectrics when stabilized in the rhombohedral perovskite structure, and which are lost in the orthorhombic modification. Stability arguments locate this interesting composition range as 0.20, but saturates at a finite value that depends on the resistance of the normal superconducting interfaces, and their distance from the path of the temperature gradient. The reduction of G(T) is determined primarily by the suppression of the density of states in the proximity-coupled normal metal along the path of the temperature gradient. G(T) is also a strongly nonlinear function of the thermal current, as found in recent experiments. PMID- 15904096 TI - Charged particles on a two-dimensional lattice subject to anisotropic Jahn-Teller interactions. AB - The properties of a system of charged particles on a 2D lattice, subject to an anisotropic Jahn-Teller-type interaction and 3D Coulomb repulsion, are investigated. In the mean-field approximation without Coulomb interaction, the system displays a phase transition of first order. When the long-range Coulomb interaction is included, Monte Carlo simulations show that the system displays very diverse mesoscopic textures, ranging from spatially disordered pairs to ordered arrays of stripes, or charged clusters, depending only on the ratio of the two interactions (and the particle density). Remarkably, charged objects with an even number of particles are more stable than with an odd number of particles. We suggest that the diverse functional behavior-including superconductivity observed in oxides can be thought to arise from the self-organization of this type. PMID- 15904097 TI - Delocalized fermions in underdoped cuprate superconductors. AB - Low-temperature heat transport was used to investigate the ground state of high purity single crystals of the lightly doped cuprate YBa2Cu3O6.33. Samples were measured with doping concentrations on either side of the superconducting phase boundary. We report the observation of delocalized fermionic excitations at zero energy in the nonsuperconducting state, which shows that the ground state of underdoped cuprates is a thermal metal. Its low-energy spectrum appears to be similar to that of the d-wave superconductor, i.e., nodal. The insulating ground state observed in underdoped La2-xSrxCuO4 is attributed to the competing spin density-wave order. PMID- 15904098 TI - Ferromagnetic fluctuation and possible triplet superconductivity in NaxCoO2.yH2O: fluctuation-exchange study of the multiorbital Hubbard model. AB - Spin and charge fluctuations and superconductivity in NaxCoO2.yH(2)O are studied based on a multiorbital Hubbard model. Tight-binding parameters are determined to reproduce the results of band calculations. By applying the fluctuation-exchange approximation, we show that the Hund's-rule coupling between the Co t(2g) orbitals causes ferromagnetic (FM) spin fluctuation. Triplet fy((y(2)-3x(2))) wave and p-wave pairings are favored by this FM fluctuation on the hole-pocket band. We propose that, in NaxCoO2.yH(2)O, the Co t(2g) orbitals and interorbital Hund's-rule coupling play important roles on the triplet pairing, and this compound can be a first example of the triplet superconductor in which the orbital degrees of freedom play substantial roles. PMID- 15904099 TI - Phase diagram of the two-channel kondo lattice model in one dimension. AB - Employing the density matrix renormalization group method and strong-coupling perturbation theory, we study the phase diagram of the SU(2)xSU(2) Kondo lattice model in one dimension. We show that, at quarter filling, the system can exist in two phases depending on the coupling strength. The weak-coupling phase is dominated by RKKY exchange correlations, while the strong-coupling phase is characterized by strong antiferromagnetic correlations of the channel degree of freedom. These two phases are separated by a quantum critical point. For conduction-band fillings of less than one-quarter, we find a paramagnetic metallic phase at weak coupling and a ferromagnetic phase at moderate to strong coupling. PMID- 15904100 TI - Spatial chemical inhomogeneity and local electronic structure of Mn-doped Ge ferromagnetic semiconductors. AB - We have investigated the chemical distributions and the local electronic structure of potential diluted magnetic semiconductor Ge0.94Mn0.06 single crystals using scanning photoelectron microscopy (SPEM), x-ray absorption spectroscopy (XAS), and photoemission spectroscopy (PES). The SPEM image shows the stripe-shaped microstructures, which arise from the chemical phase separation between the Mn-rich and Mn-depleted phases. The Mn 2p XAS shows that the Mn ions in the Mn-rich region are in the divalent high-spin Mn2+ states but that they do not form metallic Mn clusters. The Mn 3d PES spectrum exhibits a peak centered at approximately 4 eV below E(F) and the negligible spectral weight near E(F). This study suggests that the observed ferromagnetism in Ge1-xMnx arises from the phase separated Mn-rich phase. PMID- 15904101 TI - Mixed magnetic phases in (Ga,Mn)As epilayers. AB - Two different ferromagnetic-paramagnetic transitions are detected in (Ga,Mn)As/GaAs(001) epilayers from ac susceptibility measurements: transition at a higher temperature results from (Ga,Mn)As cluster phases with [110] uniaxial anisotropy and that at a lower temperature is associated with a ferromagnetic (Ga,Mn)As matrix with 100 cubic anisotropy. A change in the magnetic easy axis from [100] to [110] with increasing temperature can be explained by the reduced contribution of 100 cubic anisotropy to the magnetic properties above the transition temperature of the (Ga,Mn)As matrix. PMID- 15904102 TI - Hysteresis loops and adiabatic Landau-Zener-Stuckelberg transitions in the magnetic molecule {V6}. AB - We have observed hysteresis loops and abrupt magnetization steps in the magnetic molecule {V(6)}, where each molecule comprises a pair of identical spin triangles, in the temperature range 1-5 K for external magnetic fields B with sweep rates of several Tesla per millisecond executing a variety of closed cycles. The hysteresis loops are accurately reproduced using a generalization of the Bloch equation based on direct one-phonon transitions between the instantaneous Zeeman-split levels of the ground state (an S=1/2 doublet) of each spin triangle. The magnetization steps occur for B approximately 0, and they are explained in terms of adiabatic Landau-Zener-Stuckelberg transitions between the lowest magnetic energy levels as modified by an intertriangle anisotropic exchange of order 0.4 K. PMID- 15904103 TI - Line of critical points in 2+1 dimensions: quantum critical loop gases and non Abelian gauge theory. AB - In this Letter, we (1) construct a one-parameter family of lattice models of interacting spins; (2) obtain their exact ground states; (3) derive a statistical mechanical analogy which relates their ground states to O(n) loop gases; (4) show that the models are critical for degamma(') decay, primordial nucleosynthesis, star cooling, and other phenomena set lower limits on the scale of chirality-flip operators in the 1-15 TeV range if the operators have coefficients given by the corresponding Yukawa couplings. Simple renormalizable models induce gamma(') interactions with leptons or quarks at two loops, and may provide a cold dark matter candidate. PMID- 15904134 TI - Evidence for strange-quark contributions to the nucleon's form factors at Q2=0.108 (GeV/c)2. AB - We report on a measurement of the parity violating asymmetry in the elastic scattering of polarized electrons off unpolarized protons with the A4 apparatus at MAMI in Mainz at a four momentum transfer value of Q(2)=0.108 (GeV/c)(2) and at a forward electron scattering angle of 30 degrees p)=[-1.36+/-0.29(stat)+/-0.13(syst)]x10(-6). The expectation from the standard model assuming no strangeness contribution to the vector current is A(0)=(-2.06+/-0.14)x10(-6). We have improved the statistical accuracy by a factor of 3 as compared to our previous measurements at a higher Q2. We have extracted the strangeness contribution to the electromagnetic form factors from our data to be G(s)(E)+0.106G(s)(M)=0.071+/-0.036 at Q(2)=0.108 (GeV/c)(2). We again find the value for G(s)(E)+0.106G(s)(M) to be positive, this time at an improved significance level of two sigma. PMID- 15904136 TI - Two-nucleon transfer reactions uphold supersymmetry in atomic nuclei. AB - The spectroscopic strengths of two-nucleon transfer reactions constitute a stringent test for two-nucleon correlations in the nuclear wave functions. A set of closed analytic expressions for ratios of spectroscopic factors is derived in the framework of nuclear supersymmetry. These ratios are parameter independent and provide a direct test of the wave functions. A comparison between the recently measured 198Hg(d-->,alpha)196Au reaction and the predictions from the nuclear quartet supersymmetry lends further support to the validity of supersymmetry in nuclear physics. PMID- 15904137 TI - Precision spectroscopy and density-dependent frequency shifts in ultracold Sr. AB - By varying the density of an ultracold 88Sr sample from 10(9) to>10(12) cm(-3), we make the first definitive measurement of the density-related frequency shift and linewidth broadening of the 1S0-3P1 optical clock transition in an alkaline earth system. In addition, we report the most accurate measurement to date of the 88Sr 1S0-3P1 optical clock transition frequency. Including a detailed analysis of systematic errors, the frequency is [434 829 121 312 334+/-20(stat)+/-33(syst)] Hz. PMID- 15904138 TI - Laser cooling in an optical shaker. AB - We propose a novel generic approach to laser cooling based on the nonresonant interactions of atoms and molecules with optical standing waves experiencing sudden phase jumps. The technique, termed "optical shaking," combines the elements of stochastic cooling and Sisyphus cooling. An optical signal that measures the instantaneous force applied by the standing wave on the ensemble of particles is used as feedback to determine the phase jumps. This guarantees a drift towards lower energies and higher phase-space density without the loss of particles typical of evaporative cooling. PMID- 15904139 TI - Efficient and long-lived field-free orientation of molecules by a single hybrid short pulse. AB - We show that a combination of a half-cycle pulse and a short nonresonant laser pulse produces a strongly enhanced postpulse orientation. Robust transients that display both efficient and long-lived orientation are obtained. The mechanism is analyzed in terms of optimal oriented target states in finite Hilbert subspaces and shows that hybrid pulses can prove useful for other control issues. PMID- 15904140 TI - Isotope effects in dipole-bound anions of acetone. AB - Precision measurements using the Rydberg charge-exchange and electric field detachment methods find that the dipole-bound electron affinity (EA) of acetone (C3H6O) is 55+/-10 mueV greater than for deuterated acetone (C3D6O). The result agrees well with a theoretical prediction obtained with high-level electronic structure and anharmonic vibrational calculations. The dipole moments calculated for the vibrationally averaged structures of C3H6O and C3D6O show that the isotope effect (2% reduction) on the EA of acetone is mainly due to a slight reduction (0.5%) of the average dipole moment upon deuteration. PMID- 15904132 TI - Measurement of the WW production cross section in pp collisions at square root[s]=1.96 TeV. AB - We present a measurement of the W boson pair-production cross section in pp collisions at a center-of-mass energy of sqrt[s]=1.96 TeV. The data, collected with the Run II D0 detector at Fermilab, correspond to an integrated luminosity of 224-252 pb(-1) depending on the final state (ee, emu, or mumu). We observe 25 candidates with a background expectation of 8.1+/-0.6(stat)+/-0.6(syst)+/ 0.5(lum) events. The probability for an upward fluctuation of the background to produce the observed signal is 2.3x10(-7), equivalent to 5.2 standard deviations. The measurement yields a cross section of 13.8(+4.3)(-3.8)(stat)+1.2-0.9(syst)+/ 0.9(lum) pb, in agreement with predictions from the standard model. PMID- 15904141 TI - Electron impact ionization in the presence of a laser field: a kinematically complete (ngammae,2e) experiment. AB - Single ionization of He by 1 keV electron impact in the presence of an intense (I=4 x 10(12) W/cm(2)) laser field (lambda=1064 nm) has been explored in a kinematically complete experiment using a reaction microscope. Distinct differences in the singly to fully differential cross sections compared to the field-free situation are observed which cannot be explained by a first-order quantum calculation. Major features, such as the number of photons exchanged and the modification of the energy spectrum of emitted electrons, can be understood qualitatively within a simple classical model. PMID- 15904142 TI - Onset of superfluidity in small CO2(4He)N clusters. AB - We provide definitive theoretical evidence for the onset of superfluidity in small helium clusters doped with molecules at less than one solvation shell, with quantitative analysis of spectroscopic constants for CO2 in (4)He(N) in terms of nonclassical rotational inertia and helium superfluidity calculated by path integral methods. We find a significant superfluid response for N>/=5, with essentially unit response to rotations around the CO2 axis and partial response to rotations about an axis perpendicular to the CO2 axis for N>/=6. This anisotropic superfluid response is shown to be responsible for the N dependence of measured CO2 rotational spectra in (4)He(N). PMID- 15904143 TI - Hilbert's 17th problem and the quantumness of states. AB - A state of a quantum system can be regarded as classical (quantum) with respect to measurements of a set of canonical observables if and only if there exists (does not exist) a well defined, positive phase-space distribution, the so called Glauber-Sudarshan P representation. We derive a family of classicality criteria that requires that the averages of positive functions calculated using P representation must be positive. For polynomial functions, these criteria are related to Hilbert's 17th problem, and have physical meaning of generalized squeezing conditions; alternatively, they may be interpreted as nonclassicality witnesses. We show that every generic nonclassical state can be detected by a polynomial that is a sum-of-squares of other polynomials. We introduce a very natural hierarchy of states regarding their degree of quantumness, which we relate to the minimal degree of a sum-of-squares polynomial that detects them. PMID- 15904144 TI - Spin-dependent forces on trapped ions for phase-stable quantum gates and entangled states of spin and motion. AB - Favored schemes for trapped-ion quantum logic gates use bichromatic laser fields to couple internal qubit states with external motion through a "spin-dependent force." We introduce a new degree of freedom in this coupling that reduces its sensitivity to phase decoherence. We demonstrate bichromatic spin-dependent forces on a single trapped 111Cd+ ion, and show that phase coherence of the resulting entangled states of spin and motion depends critically upon the spectral arrangement of the optical fields. This applies directly to the operation of entangling gates on multiple ions. PMID- 15904135 TI - Search for anomalous heavy-flavor quark production in association with W bosons. AB - We search for anomalous production of heavy-flavor quark jets in association with W bosons at the Fermilab Tevatron pp Collider in final states in which the heavy flavor quark content is enhanced by requiring at least one tagged jet in an event. Jets are tagged using one algorithm based on semileptonic decays of b/c hadrons, and another on their lifetimes. We compare e+jets (164 pb(-1)) and mu+jets (145 pb(-1)) channels collected with the D0 detector at sqrt[s]=1.96 TeV to expectations from the standard model and set upper limits on anomalous production of such events. PMID- 15904145 TI - Atmospheric turbulence and orbital angular momentum of single photons for optical communication. AB - The effects of propagation through random aberrations on coherence for single photon communication systems based on orbital angular momentum states are quantified. A rotational coherence function is derived which leads to scattering equations for azimuthal modes of different orbital angular momentum states. The effect on a single-photon communication system is quantified using the channel capacity. The work shows that the decoherence effect of atmospheric turbulence on such systems is important even for weak turbulence. PMID- 15904146 TI - Tunable all-optical delays via Brillouin slow light in an optical fiber. AB - We demonstrate a technique for generating tunable all-optical delays in room temperature single-mode optical fibers at telecommunication wavelengths using the stimulated Brillouin scattering process. This technique makes use of the rapid variation of the refractive index that occurs in the vicinity of the Brillouin gain feature. The wavelength at which the induced delay occurs is broadly tunable by controlling the wavelength of the laser pumping the process, and the magnitude of the delay can be tuned continuously by as much as 25 ns by adjusting the intensity of the pump field. The technique can be applied to pulses as short as 15 ns. This scheme represents an important first step towards implementing slow light techniques for various applications including buffering in telecommunication systems. PMID- 15904147 TI - Carrier-envelope phase-controlled quantum interference in optical poling. AB - We demonstrate the efficiency of the optical poling process that depends on the CE phase-controlled quantum interference. For the experiment we employed our noncollinear optical parametric amplifier system for the self-stabilization of the CE phase, with the f-to-2f spectral interferometry system to control the CE phase. PMID- 15904148 TI - Determination of g and mu using multiply scattered light in turbid media. AB - We propose an inversion scheme to reconstruct the scattering coefficient mu and the anisotropy factor g that characterize the optical properties of a turbid medium. It is based on a theory for the scattering of light inside the medium from an angularly collimated light source. We demonstrate the feasibility of this method using light scattering data obtained from a Monte Carlo simulation. PMID- 15904149 TI - Transport of quantum noise through random media. AB - We present an experimental study of the propagation of quantum noise in a multiple scattering random medium. Both static and dynamic scattering measurements are performed: the total transmission of noise is related to the mean free path for scattering, while the noise frequency correlation function determines the diffusion constant. The quantum noise observables are found to scale markedly differently with scattering parameters compared to classical noise observables. The measurements are explained with a full quantum model of multiple scattering. PMID- 15904150 TI - Time-oscillating Lyapunov modes and the momentum autocorrelation function. AB - The Lyapunov vectors corresponding to the steps of Lyapunov spectra for many particle systems show time-oscillating behavior in two types of Lyapunov modes, one associated with time-translational invariance and the other with spatial translational invariance. Our result is that, for each coordinate direction, the longest period of the Lyapunov modes is twice as long as the period of the momentum autocorrelation function. A simple explanation for this relation is proposed and we argue that this result is generally true for many-particle systems. This gives the first quantitative connection between the Lyapunov modes and an experimentally accessible quantity. PMID- 15904151 TI - Heat conduction paradox involving second-sound propagation in moving media. AB - In this Letter, we revisit the Maxwell-Cattaneo law of finite-speed heat conduction. We point out that the usual form of this law, which involves a partial time derivative, leads to a paradoxical result if the body is in motion. We then show that by using the material derivative of the thermal flux, in lieu of the local one, the paradox is completely resolved. Specifically, that using the material derivative yields a constitutive relation that is Galilean invariant. Finally, we show that under this invariant reformulation, the system of governing equations, while still hyperbolic, cannot be reduced to a single transport equation in the multidimensional case. PMID- 15904152 TI - High energy gain of trapped electrons in a tapered, diffraction-dominated inverse free-electron laser. AB - Energy gain of trapped electrons in excess of 20 MeV has been demonstrated in an inverse-free-electron-laser (IFEL) accelerator experiment. A 14.5 MeV electron beam is copropagated with a 400 GW CO2 laser beam in a 50 cm long undulator strongly tapered in period and field amplitude. The Rayleigh range of the laser, approximately 1.8 cm, is much shorter than the undulator length yielding a diffraction-dominated interaction. Experimental results on the dependence of the acceleration on injection energy, laser focus position, and laser power are discussed. Simulations, in good agreement with the experimental data, show that most of the energy gain occurs in the first half of the undulator at a gradient of 70 MeV/m and that the structure in the measured energy spectrum arises because of higher harmonic IFEL interaction in the second half of the undulator. PMID- 15904153 TI - Particle simulation of an ultrarelativistic two-stream instability. AB - A two-stream instability in an unmagnetized plasma is examined by a particle-in cell simulation. Each beam initially consists of cold electrons and protons that stream at a relative Lorentz factor 100. This is representative for plasma close to the external shocks of gamma-ray bursts. An electrostatic wave develops which saturates by trapping electrons. This wave collapses and the resulting electrostatic turbulence gives an electron momentum distribution that resembles a power law with a spectral break. Some electrons reach Lorentz factors over 1000. PMID- 15904154 TI - Hard-sphere-like dynamics in a non-hard-sphere liquid. AB - The collective dynamics of liquid gallium close to the melting point has been studied using inelastic x-ray scattering to probe length scales smaller than the size of the first coordination shell. Although the structural properties of this partially covalent liquid strongly deviate from a simple hard-sphere model, the dynamics, as reflected in the quasielastic scattering, are beautifully described within the framework of the extended heat mode approximation of Enskog's kinetic theory, analytically derived for a hard-sphere system. The present work demonstrates, therefore, the applicability of Enskog's theory beyond simple liquids. PMID- 15904155 TI - Supersolid state of matter. AB - We prove that the necessary condition for a solid to be also a superfluid is to have zero-point vacancies, or interstitial atoms, or both, as an integral part of the ground state. As a consequence, in the absence of symmetry between vacancies and interstitials, superfluidity has a zero probability to occur in commensurate solids which break continuous translation symmetry. We discuss recent 4He experiments by Kim and Chan in the context of this theorem, question its bulk supersolid interpretation, and offer an alternative explanation in terms of superfluid interfaces. PMID- 15904156 TI - Thermally driven josephson oscillations in superfluid 4He. AB - We find that a temperature differential can drive superfluid oscillations in 4He. The oscillations are excited by a heater which causes a time dependent temperature differential across an array of 70 nm apertures. By measuring the oscillation frequency and simultaneously determining both temperature and pressure differentials we prove the validity of the most general form of the Josephson frequency relation. These observations were made near saturated vapor pressure, within a few mK of the superfluid transition temperature. PMID- 15904157 TI - Hydrogen embrittlement of aluminum: the crucial role of vacancies. AB - We report first-principles calculations which demonstrate that vacancies can combine with hydrogen impurities in bulk aluminum and play a crucial role in the embrittlement of this prototypical ductile solid. Our studies of hydrogen-induced vacancy superabundant formation and vacancy clusterization in aluminum lead to the conclusion that a large number of H atoms (up to 12) can be trapped at a single vacancy, which overcompensates the energy cost to form the defect. In the presence of trapped H atoms, three nearest-neighbor single vacancies which normally would repel each other, aggregate to form a trivacancy on the slip plane of Al, acting as embryos for microvoids and cracks and resulting in ductile rupture along these planes. PMID- 15904158 TI - In situ observation of thermal relaxation of interstitial-vacancy pair defects in a graphite gap. AB - Direct observation of individual defects during formation and annihilation in the interlayer gap of double-wall carbon nanotubes (DWNT) is demonstrated by high resolution transmission electron microscopy. The interlayer defects that bridge two adjacent graphen layers in DWNT are stable for a macroscopic time at the temperature below 450 K. These defects are assigned to a cluster of one or two interstitial-vacancy pairs (I-V pairs) and often disappear just after their formation at higher temperatures due to an instantaneous recombination of the interstitial atom with vacancy. Systematic observations performed at the elevated temperatures find a threshold for the defect annihilation at 450-500 K, which, indeed, corresponds to the known temperature for the Wigner energy release. PMID- 15904159 TI - Kinetic pathway for the formation of fe nanowires on stepped Cu111 surfaces. AB - We report the discovery of a novel kinetic pathway for the formation of one dimensional Fe nanowires of single atom width on stepped Cu(111) surfaces. This pathway, identified through extensive total-energy calculations within density functional theory, establishes that the stable structure involves a row of Fe atoms on the upper edge of a step. The formation of the surface wire is preceded by facile incorporation of an initial row of Fe atoms into the surface layer at one lateral lattice constant away from the step edge, which then acts as an attractor for the second exposed row of atoms. The resulting wire structure provides a natural interpretation of existing experimental results. We also explore the applicability of this mechanism in the formation of other related systems. PMID- 15904160 TI - Hydrogen storage in novel organometallic buckyballs. AB - Transition metal (TM) atoms bound to fullerenes are proposed as adsorbents for high density, room temperature, ambient pressure storage of hydrogen. C60 or C48B12 disperses TMs by charge transfer interactions to produce stable organometallic buckyballs (OBBs). A particular scandium OBB can bind as many as 11 hydrogen atoms per TM, ten of which are in the form of dihydrogen that can be adsorbed and desorbed reversibly. In this case, the calculated binding energy is about 0.3 eV/H(2), which is ideal for use on board vehicles. The theoretical maximum retrievable H2 storage density is approximately 9 wt %. PMID- 15904161 TI - Origin of the opalescence at the alpha-beta transition of quartz: role of the incommensurate phase studied by synchrotron radiation. AB - The origin of the light scattering observed at the alpha-beta transition of quartz is still a subject of controversy. We present structural studies performed during the coexistence of the alpha and the intermediate incommensurate (inc) phases using simultaneously synchrotron x-ray diffraction and optical techniques. The small and large angle light scatterings are due, respectively, to the orientation domains of the 3q inc phase and to the alpha phase twins revealed by diffuse x-ray scattering. In the vicinity of the interphase boundary, the two light scattering regions, both with perturbed properties, form a complex multiscale structure. PMID- 15904162 TI - Ergodic to nonergodic transition in liquids with a local order: the case of m toluidine. AB - The dynamic structure factor of m-toluidine has been measured by inelastic x-ray scattering in the mesoscopic Q range between 1 and 10 nm(-1), where a prepeak is revealed in the static structure factor resulting from the existence of hydrogen bonded, nanometer size clusters. Evidence is given of (i) a square-root cusp in the nonergodicity factor and of (ii) critical nonergodicity parameters which oscillate in phase with the static structure factor. These results demonstrate that local order in a liquid can coexist with the signatures of the ergodic to nonergodic transition predicted by the mode coupling theory for simple, dense liquids. PMID- 15904163 TI - Marginal fermi liquid theory in the Hubbard model. AB - We find marginal-Fermi-liquid- (MFL) like behavior in the Hubbard model on a square lattice for a range of hole doping and on-site interaction parameter U. Thereby we use a self-consistent projection operator method. It enables us to compute the momentum and frequency dependence of the single-particle excitations with high resolution. The Fermi surface is found to be holelike in the underdoped regime and electronlike in the overdoped regime. Our calculations concern normal state properties of the system. When a comparison is possible, we find consistency with finite temperature quantum Monte Carlo results. We also find a discontinuous change with doping concentration from a MFL to a Fermi-liquid behavior resulting from a collapse of the lower Hubbard band. This renders Luttinger's theorem inapplicable in the underdoped regime. PMID- 15904164 TI - Orbitally induced Peierls state in spinels. AB - We consider the superstructures, which can be formed in spinels containing on B sites the transition-metal ions with partially filled t(2g) levels. We show that, when such systems are close to the itinerant state (e.g., have an insulator-metal transition), there may appear in them an orbitally driven Peierls state. We explain by this mechanism the very unusual superstructures observed in CuIr2S4 (octamers) and MgTi2O4 (chiral superstructures) and suggest that a similar phenomenon should be observed in NaTiO2 and possibly in some other systems. PMID- 15904165 TI - Noise-enabled precision measurements of a duffing nanomechanical resonator. AB - We report quantitative measurements of the nonlinear response of a radio frequency mechanical resonator with a very high quality factor. We measure the noise-free transitions between the two basins of attraction that appear in the nonlinear regime, and find good agreement with theory. We measure the transition rate response to controlled levels of white noise, and extract the basin activation energy. This allows us to obtain precise values for the relevant frequencies and the cubic nonlinearity in the Duffing oscillator, with applications to parametric sensing. PMID- 15904166 TI - Competition between antiferromagnetism and superconductivity in high-Tc cuprates. AB - Using variational cluster perturbation theory we study the competition between d wave superconductivity (dSC) and antiferromagnetism (AF) in the t-t(')-t('')-U Hubbard model. Large scale computer calculations reproduce the overall ground state phase diagram of the high-temperature superconductors as well as the one particle excitation spectra for both hole and electron doping. We identify clear signatures of the Mott gap as well as of AF and of dSC that should be observable in photoemission experiments. PMID- 15904167 TI - Thermal transport in the hidden-order state of URu2Si2. AB - We present a study of thermal conductivity in the normal state of the heavy fermion superconductor URu2Si2. Ordering at 18 K leads to a steep increase in thermal conductivity and (in contrast with all other cases of magnetic ordering in heavy-fermion compounds) to an enhancement of the Lorenz number. By linking this observation to several other previously reported features, we conclude that most of the carriers disappear in the ordered state and this leads to a drastic increase in both the phononic and electronic mean free path. PMID- 15904168 TI - Unifying model for several classes of two-dimensional phase transition. AB - A relatively simple and physically transparent model based on quantum percolation and dephasing is employed to construct a global phase diagram which encodes and unifies the critical physics of the quantum Hall, "two-dimensional metal insulator," classical percolation and, to some extent, superconductor-insulator transitions. Using real-space renormalization group techniques, crossover functions between critical points are calculated. The critical behavior around each fixed point is analyzed and some experimentally relevant puzzles are addressed. PMID- 15904169 TI - Interacting fermions in two dimensions: beyond the perturbation theory. AB - We consider a system of 2D fermions with a short-range interaction. A straightforward perturbation theory is shown to be ill defined even for an infinitesimally weak interaction, as the perturbative series for the self-energy diverges near the mass shell. We show that the divergences result from the interaction of fermions with the zero-sound collective mode. By resumming the most divergent diagrams, we obtain a closed form of the self-energy near the mass shell. The spectral function exhibits a threshold feature at the onset of the emission of the zero-sound waves. We also show that the interaction with the zero sound does not affect a nonanalytic, T2 part of the specific heat. PMID- 15904170 TI - Hole dynamics in spin and orbital ordered vanadium perovskites. AB - A theory of doped perovskite vanadates with spin and orbital orders is presented. Mobile holes are strongly renormalized by spin excitations (magnons) in the spin G-type and orbital C-type (SG-OC) order, and orbital excitations (orbitons) in the spin C-type and orbital G-type (SC-OG) one. Hole dynamics in a staggered t(2g) orbital array is distinguished from that in the antiferromagnetic order and the e(g) orbital one. The fragile character of the (SG-OC) order in Y1-xCaxVO3 is attributed to the orbiton softening induced by a reduction of the spin order parameter. PMID- 15904171 TI - Field theory for the global density of states distribution function of disordered conductors. AB - A field-theoretical representation is suggested for the electron global density of states distribution function P(nu) in extended disordered conductors. This opens a way to study the complete statistics of fluctuations. The approach is based on a functional integration over bilocal functions Psir(1)(r(2)) instead of the integration over local functions in the usual functional representation for moments of physical quantities. The formalism allows one to perform the disorder averaging and to derive an analog of the usual nonlinear sigma model-a slow functional of a supermatrix field Qr(1)(r(2))(r) approximately Psi(rr(1)) composite functionPsi (r(2)r). As an application of the formalism, the long-tail asymptotics of P(nu) is derived. PMID- 15904172 TI - First-principles study of the switching mechanism of [2]catenane molecular electronic devices. AB - We present a first-principles study of the coherent charge transport properties of bistable [2]catenane molecular monolayers sandwiched between Au(111) electrodes. We find that conduction channels around the Fermi level are dominated by the two highest occupied molecular orbital levels from tetrathiafulvalene (TTF) and dioxynaphthalene (DNP) and the two lowest unoccupied molecular orbital levels from tetracationic cyclophane (CBPQT4+), and the OFF to ON switching results from the energetic shifts of these orbitals as CBPQT4+ moves from TTF to DNP. We show that the superposition principle can be adopted for predicting the function of the composite device. PMID- 15904173 TI - Electronic transport spectroscopy of carbon nanotubes in a magnetic field. AB - We report magnetic field spectroscopy measurements in carbon nanotube quantum dots exhibiting fourfold shell structure in the energy level spectrum. The magnetic field induces a large splitting between the two orbital states of each shell, demonstrating their opposite magnetic moment and determining transitions in the spin and orbital configuration of the quantum dot ground state. We use inelastic cotunneling spectroscopy to accurately resolve the spin and orbital contributions to the magnetic moment. A small coupling is found between orbitals with opposite magnetic moment leading to anticrossing behavior at zero field. PMID- 15904174 TI - Dissipation-driven phase transition in two-dimensional Josephson arrays. AB - We analyze the interplay of dissipative and quantum effects in the proximity of a quantum phase transition. The prototypical system is a resistively shunted two dimensional Josephson junction array, studied by means of an advanced Fourier path-integral Monte Carlo algorithm. The reentrant superconducting-to-normal phase transition driven by quantum fluctuations, recently discovered in the limit of infinite shunt resistance, persists for moderate dissipation strength but disappears in the limit of small resistance. For large quantum coupling our numerical results show that, beyond a critical dissipation strength, the superconducting phase is always stabilized at sufficiently low temperature. Our phase diagram explains recent experimental findings. PMID- 15904175 TI - Evidence for charge Kondo effect in superconducting Tl-doped PbTe. AB - We report results of low-temperature thermodynamic and transport measurements of Pb1-xTlxTe single crystals for Tl concentrations up to the solubility limit of approximately x=1.5%. For all doped samples, we observe a low-temperature resistivity upturn that scales in magnitude with the Tl concentration. The temperature and field dependence of this upturn are consistent with a charge Kondo effect involving degenerate Tl valence states differing by two electrons, with a characteristic Kondo temperature T(K) approximately 6 K. The observation of such an effect supports an electronic pairing mechanism for superconductivity in this material and may account for the anomalously high T(c) values. PMID- 15904176 TI - Superconductivity in charge Kondo systems. AB - We present a theory for superconductivity and charge Kondo fluctuations, i.e., resonant quantum valence fluctuations by two charge units, for Tl-doped PbTe. We show that Tl is very special as it first supplies a certain amount of charge carriers to the PbTe-valence band and then puts itself into a self-tuned resonant state to yield a new, robust pairing mechanism for these carriers. PMID- 15904177 TI - Using Josephson vortex lattices to control terahertz radiation: tunable transparency and terahertz photonic crystals. AB - The Josephson vortex (JV) lattice is a periodic array that scatters electromagnetic waves in the THz-frequency range. We show that JV lattices can produce a photonic band-gap structure (THz photonic crystal) with easily tunable forbidden zones controlled by the in-plane magnetic field. The scattering of electromagnetic waves by JVs results in a strong magnetic-field dependence of the reflection and transparency. Fully transparent or fully reflected frequency windows can be conveniently tuned by the in-plane magnetic field. These proposals are potentially useful for controllable THz filters. PMID- 15904178 TI - Temperature dependence of the superconducting gap in high-Tc cuprates. AB - It is proposed that the temperature dependence of the superconducting gap Delta(T) in high-T(c) cuprates can be predicted just from the knowledge of Delta(0) and the critical temperature T(c), and, in particular, Delta(0)/T(c)>4 implies that Delta(T(c)) not equal 0, while Delta(0)/T(c) ue- nu interactions. When cast into limits on n --> pe- nu coupling constants, our results yield constraints on scalar and tensor weak interactions improved by more than an order of magnitude over the current experimental limits. When combined with the existing limits, our results yield absolute value(C(S)/C(V)) approximately < 5 x 10(-3), absolute value(C'(S)/C(V)) approximately < 5 x 10(-3), absolute value(C(T)/C(A)) approximately < 1.2 x 10( 2), and absolute value(C'(T)/C(A)) approximately < 1.2 x 10(-2). PMID- 15904215 TI - D(s0)+ (2317)-D0(2308) mass difference as evidence for tetraquarks. AB - We argue that the anomalously small mass difference between the D(s0)+ (2317) and the recently observed D0(2308) (Belle) state is the first "smoking gun" experimental evidence of their tetraquark [cq(q q)] structure. The recently reported D(sJ)+ (2632) (SELEX) state completes the low-lying nonexotic tetraquark spectrum, as predicted by 't Hooft's instanton-induced effective quark interaction. PMID- 15904214 TI - Measurement of helicity-dependent photoabsorption cross sections on the neutron from 815 to 1825 MeV. AB - Helicity-dependent total photoabsorption cross sections on the deuteron have been measured for the first time at ELSA (Bonn) in the photon energy range from 815 to 1825 MeV. Circularly polarized tagged photons impinging on a longitudinally polarized LiD target have been used together with a highly efficient 4pi detector system. The data around 1 GeV are not compatible with predictions from existing multipole analyses. From the measured energy range an experimental contribution to the GDH integral on the neutron of [33.9 +/- 5.5(stat) +/- 4.5(syst)] microb is extracted. PMID- 15904217 TI - 29Na: defining the edge of the island of inversion for Z=11. AB - The low-energy level structure of the exotic Na isotopes (28,29)Na has been investigated through beta-delayed gamma spectroscopy. The N=20 isotones for Z=10 12 are considered to belong to the "island of inversion" where intruder configurations dominate the ground state wave function. However, it is an open question as to where and how the transition from normal to intruder dominated configurations happens in an isotopic chain. The present work, which presents the first detailed spectroscopy of (28,29)Na, clearly demonstrates that such a transition in the Na isotopes occurs between 28Na (N=17) and 29Na (N=18), supporting the smaller N=20 shell gap in neutron-rich sd shell nuclei. The evidence for inverted shell structure is found in beta-decay branching ratios, intruder dominated spectroscopy of low-lying states, and shell model analysis. PMID- 15904216 TI - Charged meson rapidity distributions in central Au+Au collisions at square root(sNN) = 200 GeV. AB - We have measured rapidity densities dN/dy of pi+/- and K+/- over a broad rapidity range (-0.1 < y < 3.5) for central Au + Au collisions at square root(sNN) = 200 GeV. These data have significant implications for the chemistry and dynamics of the dense system that is initially created in the collisions. The full phase space yields are 1660 +/- 15 +/- 133 (pi+), 1683 +/- 16 +/- 135 (pi-), 286 +/- 5 +/- 23 (K+), and 242 +/- 4 +/- 19 (K-). The systematics of the strange to nonstrange meson ratios are found to track the variation of the baryochemical potential with rapidity and energy. Landau-Carruthers hydrodynamics is found to describe the bulk transport of the pions in the longitudinal direction. PMID- 15904218 TI - Deuteron transfer in N=Z nuclei. AB - Predictions are obtained for T=0 and T=1 deuteron-transfer intensities between self-conjugate N=Z nuclei on the basis of a simplified interacting boson model which considers bosons without orbital angular momentum but with full spin isospin structure. These transfer predictions can be correlated with nuclear binding energies in specific regions of the mass table. PMID- 15904211 TI - Measurement of the ratio of inclusive cross sections sigma(pp --> Z + b jet)/sigma(pp --> Z + jet) at square root(s) = 1.96 TeV. AB - Using the data collected with the D0 detector at square root(s) = 1.96 TeV, for integrated luminosities of about 180 pb(-1), we have measured the ratio of inclusive cross sections for pp --> Z + b jet to pp --> Z + jet production. The inclusive Z + b-jet reaction is an important background to searches for the Higgs boson in associated ZH production at the Fermilab Tevatron collider. Our measurement is the first of its kind, and relies on the Z --> e+ e- and Z --> mu+ mu- modes. The combined measurement of the ratio yields 0.021+/-0.005 for hadronic jets with transverse momenta pT > 20 GeV/c and pseudorapidities absolute value(eta) < 2.5, consistent with next-to-leading-order predictions of the standard model. PMID- 15904219 TI - Isotopic scaling and the symmetry energy in spectator fragmentation. AB - Isotopic effects in the fragmentation of excited target residues following collisions of 12C on (112,124)Sn at incident energies of 300 and 600 MeV per nucleon were studied with the INDRA 4pi detector. The measured yield ratios for light particles and fragments with atomic number Z < or = 5 obey the exponential law of isotopic scaling. The deduced scaling parameters decrease strongly with increasing centrality to values smaller than 50% of those obtained for the peripheral event groups. Symmetry-term coefficients, deduced from these data within the statistical description of isotopic scaling, are near gamma = 25 MeV for peripheral and gamma < 15 MeV for central collisions. PMID- 15904220 TI - Measurement of the (199)Hg+ 5d9 6s2 (2)D(5/2) electric quadrupole moment and a constraint on the quadrupole shift. AB - The electric-quadrupole moment of the (199)Hg+ 5d9 6s2 (2)D(5/2) state is measured to be theta(D,5/2) = -2.29(8) x 10(-40) C m2. This value was determined by measuring the frequency of the (199)Hg+ 5d10 6s (2)S(1/2) --> 5d9 6s2 (2)D(5/2) optical clock transition for different applied electric-field gradients. An isolated, mechanically stable optical cavity provides a frequency reference for the measurement. We compare the results with theoretical calculations and discuss the implications for the accuracy of an atomic clock based upon this transition. We now expect that the frequency shift caused by the interaction of the quadrupole moment with stray electric-field gradients will not limit the accuracy of the Hg+ optical clock at the 10(-18) level. PMID- 15904221 TI - Intense-laser-field ionization of the hydrogen molecular ions H2+ and D2+ at critical internuclear distances. AB - Fragmentation of H2+ and D2+ in ion beams has been studied with short intense laser pulses (100 fs, I=5x10(13)-1x10(15) W/cm2) and by a high-resolution two dimensional velocity imaging technique. In the Coulomb explosion channel, at intensities just above the threshold for this process, we observe a clear structure in the kinetic energy spectra not previously found or predicted. The peaks can be attributed to single vibrational levels. We interpret this observation as a dissociative allocation of the electron to a proton followed by enhanced ionization at a well-defined "critical" overstretched internuclear distance. When using longer pulses we observe three separate Coulomb explosion velocity groups corresponding to critical distances of about 8, 11, and 15 a.u. PMID- 15904222 TI - Direct observation of distorted wave effects in ethylene using the (e,2e) reaction. AB - We report here the direct measurements of electron momentum distributions for ethylene using the (e,2e) reaction at different impact energies from 400 to 2400 eV. The "turn up" effects in the (e,2e) cross sections of the 1b(3g) orbital compared with the plane-wave impulse approximation calculations were observed at low and high momentum regions, and such discrepancies become smaller with the increase of the impact electron energies. It is suggested that the observed discrepancies are due to the distorted-wave effects in molecules, while appropriate theoretical calculations using distorted waves in molecules could not be achieved until now. PMID- 15904223 TI - Electron-impact excitation out of the metastable levels of krypton. AB - We have measured the electron-impact excitation cross sections out of the two metastable levels of Kr into the ten levels of the 4p(5)5p configuration. For a common 4p(5)5p final level, the peak excitation cross sections out of the two individual 4p(5)5s metastable levels are found to differ by 1 to 2 orders of magnitude. This is explained by the special features of the electronic structure of the two configurations involved. The peak cross sections are 10 to 1600 times larger than the corresponding peak cross sections out of the ground state. PMID- 15904213 TI - Improved measurement of CP asymmetries in B0 --> (cc)K(0(*)) decays. AB - We present results on time-dependent CP asymmetries in neutral B decays to several CP eigenstates. The measurements use a data sample of about 227 x 10(6) upsilon(4S) --> BB decays collected by the BABAR detector at the PEP-II asymmetric-energy B Factory at SLAC. The amplitude of the CPasymmetry, sin2beta in the standard model, is derived from decay-time distributions from events in which one neutral B meson is fully reconstructed in a final state containing a charmonium meson and the other B meson is determined to be either a B0 or B0 from its decay products. We measure sin2beta = 0.722 +/- 0.040(stat) +/- 0.023(syst) in agreement with the standard model expectation. PMID- 15904224 TI - Radiation field and resonant two-center dielectronic transitions in relativistic ion-atom collisions. AB - We consider projectile-electron excitation and loss in relativistic collisions of ionic projectiles with excited atoms. We show that under certain conditions electron transitions in the ion and atom can be resonantly coupled in the collision via the radiation field. The resonance becomes possible due to the Doppler effect, has a well-defined impact energy threshold, and clearly manifests itself in the cross sections. Since the range of the ion-atom interaction in the resonance case is very long, the presence of other atoms in the target medium as well as the size of the space occupied by the medium have to be taken into account. As a result, the cross section may become dependent on the density of the target atoms and/or the target size. PMID- 15904225 TI - Instability and entanglement of the ground state of the Dicke model. AB - Using tools of quantum information theory we show that the ground state of the Dicke model exhibits an infinite sequence of instabilities (quantum-phase-like transitions). These transitions are characterized by abrupt changes of the bi partite entanglement between atoms at critical values kappa(j) of the atom-field coupling parameter kappa and are accompanied by discontinuities of the first derivative of the energy of the ground state. We show that in a weak-coupling limit (kappa1 < or = kappa < or = kappa2) the Coffman-Kundu-Wootters inequalities are saturated, which proves that for these values of the coupling no intrinsic multipartite entanglement (neither among the atoms nor between the atoms and the field) is generated by the atom-field interaction. We show that in the strong coupling limit the entangling interaction with atoms leads to a highly sub Poissonian photon statistics of the field mode. PMID- 15904226 TI - Synchronization of delay-coupled oscillators: a study of semiconductor lasers. AB - Two delay-coupled semiconductor lasers are studied in the regime where the coupling delay is comparable to the time scales of the internal laser oscillations. Detuning the optical frequency between the two lasers, novel delay induced scenarios leading from optical frequency locking to successive states of periodic intensity pulsations are observed. We demonstrate and analyze these dynamical phenomena experimentally using two distinct laser configurations. A theoretical treatment reveals the universal character of our findings for delay coupled systems. PMID- 15904227 TI - Brillouin zone spectroscopy of nonlinear photonic lattices. AB - We present a novel, real-time, experimental technique for linear and nonlinear Brillouin zone spectroscopy of photonic lattices. The method relies on excitation with random-phase waves and far-field visualization of the spatial spectrum of the light exiting the lattice. Our technique facilitates mapping the borders of the extended Brillouin zones and the areas of normal and anomalous dispersion within each zone. For photonic lattices with defects (e.g., photonic crystal fibers), our technique enables far-field visualization of the defect mode overlaid on the extended Brillouin zone structure of the lattice. The technique is general and can be used for photonic crystal fibers as well as for periodic structures in areas beyond optics. PMID- 15904228 TI - Quantum ratchets in dissipative chaotic systems. AB - Using the method of quantum trajectories, we study a quantum chaotic dissipative ratchet appearing for particles in a pulsed asymmetric potential in the presence of a dissipative environment. The system is characterized by directed transport emerging from a quantum strange attractor. This model exhibits, in the limit of small effective Planck constant, a transition from quantum to classical behavior, in agreement with the correspondence principle. We also discuss parameter values suitable for the implementation of the quantum ratchet effect with cold atoms in optical lattices. PMID- 15904229 TI - Effective desynchronization by nonlinear delayed feedback. AB - We show that nonlinear delayed feedback opens up novel means for the control of synchronization. In particular, we propose a demand-controlled method for powerful desynchronization, which does not require any time-consuming calibration. Our technique distinguishes itself by its robustness against variations of system parameters, even in strongly coupled ensembles of oscillators. We suggest our method for mild and effective deep brain stimulation in neurological diseases characterized by pathological cerebral synchronization. PMID- 15904230 TI - Modeling of wave propagation in inhomogeneous media. AB - We present a methodology providing a new perspective on modeling and inversion of wave propagation satisfying time-reversal invariance and reciprocity in generally inhomogeneous media. The approach relies on a representation theorem of the wave equation to express the Green function between points in the interior as an integral over the response in those points due to sources on a surface surrounding the medium. Following a predictable initial computational effort, Green's functions between arbitrary points in the medium can be computed as needed using a simple cross-correlation algorithm. PMID- 15904231 TI - Mechanism for flow-rate controlled breakup in confined geometries: a route to monodisperse emulsions. AB - This Letter describes a quasistationary breakup of an immiscible, inviscid fluid at low capillary numbers. The breakup proceeds in a coflowing, viscous liquid, in a confined geometry of a long and narrow orifice. In contrast to the capillary instability in an unbounded fluid, the collapse proceeds through a series of equilibria, each yielding the minimum interfacial energy of the fluid-fluid interface. The process is slow in comparison to typical relaxation speeds of the interface, and it is reversible. Its quasistatic character of collapse forms the basis for controlled, high-throughput generation of monodisperse fluid dispersions. PMID- 15904232 TI - Numerical study of dynamo action at low magnetic Prandtl numbers. AB - We present a three-pronged numerical approach to the dynamo problem at low magnetic Prandtl numbers P(M). The difficulty of resolving a large range of scales is circumvented by combining direct numerical simulations, a Lagrangian averaged model and large-eddy simulations. The flow is generated by the Taylor Green forcing; it combines a well defined structure at large scales and turbulent fluctuations at small scales. Our main findings are (i) dynamos are observed from P(M)=1 down to P(M)=10(-2), (ii) the critical magnetic Reynolds number increases sharply with P(M)(-1) as turbulence sets in and then it saturates, and (iii) in the linear growth phase, unstable magnetic modes move to smaller scales as P(M) is decreased. Then the dynamo grows at large scales and modifies the turbulent velocity fluctuations. PMID- 15904233 TI - Shearing interferometer for quantifying the coherence of hard x-ray beams. AB - We report a quantitative measurement of the full transverse coherence function of the 14.4 keV x-ray radiation produced by an undulator at the Swiss Light Source. An x-ray grating interferometer consisting of a beam splitter phase grating and an analyzer amplitude grating has been used to measure the degree of coherence as a function of the beam separation out to 30 microm. Importantly, the technique provides a model-free and spatially resolved measurement of the complex coherence function and is not restricted to high resolution detectors and small fields of view. The spatial characterization of the wave front has important applications in discovering localized defects in beam line optics. PMID- 15904234 TI - Construction of exact plasma equilibrium solutions with different geometries. AB - Infinite families of exact isotropic and anisotropic plasma equilibria with and without dynamics can be constructed in different geometries, using the representation of the static MHD equilibrium system in coordinates connected with magnetic surfaces. A sample equilibrium anisotropic model of Earth magnetosheath plasma is given. PMID- 15904235 TI - Quasiparticle approach to the modulational instability of drift waves coupling to zonal flows. AB - The interaction between broadband drift mode turbulence and zonal flows has been studied through the wave-kinetic approach. Simulations have been conducted in which a particle-in-cell representation is used for the quasiparticles, while a fluid model is employed for the plasma. The interactions have been studied in a plasma edge configuration which has applications in both tokamak physics and magnetopause boundary layer studies. Simulation results show the development of a zonal flow through the modulational instability of the drift wave distribution, as well as the existence of solitary zonal flow structures about an ion gyroradius wide, drifting towards steeper relative density gradients. PMID- 15904236 TI - Laser acceleration of ion bunches at the front surface of overdense plasmas. AB - The acceleration of ions in the interaction of high intensity laser pulses with overdense plasmas is investigated with particle-in-cell simulations. For circular polarization of the laser pulses, high-density ion bunches moving into the plasma are generated at the laser-plasma interaction surface. A simple analytical model accounts for the numerical observations and provides scaling laws for the ion bunch energy and generation time as a function of pulse intensity and plasma density. PMID- 15904237 TI - Novel mechanism of anomalous electron heat conductivity and thermal crashes during Alfvenic activity in the Wendelstein 7-AS stellarator. AB - Enhanced plasma heat conductivity in the presence of kinetic Alfven waves (KAW) is predicted theoretically. The enhancement is shown to be strongest when the electron collision frequency exceeds the particle transit frequency in the wave field. Alfven waves (both KAW and ideal MHD Alfven eigenmodes generating the KAW) are studied in a shot of the Wendelstein 7-AS stellarator. On the basis of these results, strong thermal crashes observed during bursting Alfvenic activity in the mentioned shot are explained. PMID- 15904238 TI - Superfluid interfaces in quantum solids. AB - One scenario for the nonclassical moment of inertia of solid 4He discovered by Kim and Chan [Nature (London) 427, 225 (2004)] is the superfluidity of microcrystallite interfaces. On the basis of the most simple model of a quantum crystal--the checkerboard lattice solid--we show that the superfluidity of interfaces between solid domains can exist in a wide range of parameters. At strong enough interparticle interaction, a superfluid interface becomes an insulator via a quantum phase transition. Under the conditions of particle-hole symmetry, the transition is of the standard U(1) universality class in 3D, while in 2D the onset of superfluidity is accompanied by the interface roughening, driven by fractionally charged topological excitations. PMID- 15904239 TI - Vacancy-impurity complexes in highly Sb-doped Si grown by molecular beam epitaxy. AB - Positron annihilation measurements, supported by first-principles electron structure calculations, identify vacancies and vacancy clusters decorated by 1-2 dopant impurities in highly Sb-doped Si. The concentration of vacancy defects increases with Sb doping and contributes significantly to the electrical compensation. Annealings at low temperatures of 400-500 K convert the defects to larger complexes where the open volume is neighbored by 2-3 Sb atoms. This behavior is attributed to the migration of vacancy-Sb pairs and demonstrates at atomic level the metastability of the material grown by epitaxy at low temperature. PMID- 15904240 TI - Atomistic mechanisms of fatigue in nanocrystalline metals. AB - We investigate the mechanisms of fatigue behavior in nanocrystalline metals at the atomic scale using empirical force laws and molecular level simulations. A combination of molecular statics and molecular dynamics was used to deal with the time scale limitations of molecular dynamics. We show that the main atomistic mechanism of fatigue crack propagation in these materials is the formation of nanovoids ahead of the main crack. The results obtained for crack advance as a function of stress intensity amplitude are consistent with experimental studies and a Paris law exponent of about 2. PMID- 15904241 TI - Understanding of the phase transformation from fullerite to amorphous carbon at the microscopic level. AB - We study the shock-induced phase transformation from fullerite to a dense amorphous carbon phase by tight-binding molecular dynamics. For increasing hydrostatic pressures P, the C60 cages are found to polymerize at P<10 GPa, to break at P approximately 40 GPa, and to slowly collapse further at P>40 GPa. By contrast, in the presence of additional shear stresses, the cages are destroyed at much lower pressures (P<30 GPa). We explain this fact in terms of a continuum model, the snap-through instability of a spherical shell. Surprisingly, the relaxed high-density structures display no intermediate-range order. PMID- 15904242 TI - Shear strain in Nd0.5Ca0.5MnO3 at high pressures. AB - High-pressure x-ray powder diffraction has been measured on the half doped rare earth manganite Nd0.5Ca0.5MnO3 up to a pressure of 15 GPa. We report the presence of a quantifiable amount of shear distortion of the MnO6 octahedra in Nd0.5Ca0.5MnO3 at high pressures. The lattice strain of Nd0.5Ca0.5MnO3 is minimal at a crossover pressure of p* approximately 7 GPa, with the same lattice strain above and below this pressure achieved by shear and Jahn-Teller-type distortions, respectively. The increase in shear strain with increasing pressure provides a mechanism for the insulating behavior of manganites at high pressures that has not been considered before. PMID- 15904243 TI - Localization of elastic waves in heterogeneous media with off-diagonal disorder and long-range correlations. AB - Using the Martin-Siggia-Rose method, we study propagation of acoustic waves in strongly heterogeneous media which are characterized by a broad distribution of the elastic constants. Gaussian-white distributed elastic constants, as well as those with long-range correlations with nondecaying power-law correlation functions, are considered. The study is motivated in part by a recent discovery that the elastic moduli of rock at large length scales may be characterized by long-range power-law correlation functions. Depending on the disorder, the renormalization group (RG) flows exhibit a transition to localized regime in any dimension. We have numerically checked the RG results using the transfer-matrix method and direct numerical simulations for one- and two-dimensional systems, respectively. PMID- 15904244 TI - Effects of entanglement on the scattering intensity of neutron compton scattering by a proton pair in solids. AB - We calculate scattering functions S(q-->,omega) of neutron Compton scattering (NCS) for a proton (deuteron) pair in the entangled state caused by the indistinguishability of particles. From the calculation, it is concluded that the effect of entanglement on the scattering intensity is undetectable in NCS experiments because of their high energy transfer and broad energy resolution. PMID- 15904245 TI - Structure of B2O3 Glass at High Pressure: A 11B Solid-State NMR Study. AB - We report spectroscopic evidence for the pressure-induced structural changes in B2O3 glass quenched from melts at pressures up to 6 GPa using solid-state NMR. While all borons are tri-coordinated at 1 atm, the fraction of tetra-coordinated boron increases with pressure, being about 5% and 27% in the B2O3 glass quenched from melts at 2 and 6 GPa, respectively. The fraction of boroxol ring species increases with pressure up to 2 GPa and apparently decreases with further compression up to 6 GPa. Two densification mechanisms are proposed to explain the variation of boron species with pressure. PMID- 15904246 TI - Influence of chain length on the alpha-beta bifurcation in oligomeric glass formers. AB - A series of oligo(propylene glycol) dimethyl ethers has been investigated using dielectric spectroscopy in order to relate features of the glass transition dynamics to the number of monomer units N in the chain. The results show that (i) when scaled with the glass transition temperature, the beta relaxations systematically become faster for larger N whereas the alpha relaxations display nearly identical temperature evolutions, i.e., the alpha-beta bifurcation shifts towards shorter times for larger N, (ii) the bifurcation scenario displays a crossover in behavior at N approximately 10 monomer units, signaling the transition from oligomeric to polymeric behavior, and (iii) the beta relaxation has a cooperative nature. PMID- 15904247 TI - Ferryl (Fe=O) termination of the hematite alpha-Fe2O3(0001) surface. AB - Using scanning tunneling microscopy and infrared reflection absorption spectroscopy we have observed that the alpha-Fe2O3(0001) surface exhibits ferryl (Fe=O) groups, which may coexist with domains of the Fe-terminated surface. We therefore fully support ab initio calculations recently reported in the literature [W. Bergmeyer, H. Schweiger, and E. Wimmer, Phys. Rev. B 69, 195409 (2004)]. The close similarity to the results on the (0001) surfaces of Cr2O3 and V2O3 strongly suggests that the M=O termination under certain oxygen pressure conditions is the most stable for the close-packed surfaces of transition metal oxides with the corundum structure. PMID- 15904248 TI - Triplon modes of puddles. AB - Free fluctuations of the contact line of large drops ("puddles") of wavelength lambda > kappa(-1), the capillary length, cannot be seen on a solid substrate because even a small but finite hysteresis is enough to block these slow modes. We show here that vertical vibrations of the substrate (at frequency omegaE, acceleration Lambda) above a threshold amplitude Lambda(c) release the line and excite contour oscillations (triplons). We observe harmonic modes and parametric excitations at omegaE/2. We construct the phase diagram (Lambda, omegaE) of these subharmonic modes and we study their growth dynamics: they slow down near the threshold of the contour instability. PMID- 15904249 TI - Scaling of local slopes, conservation laws, and anomalous roughening in surface growth. AB - We argue that symmetries and conservation laws greatly restrict the form of the terms entering the long wavelength description of growth models exhibiting anomalous roughening. This is exploited to show by dynamic renormalization group arguments that intrinsic anomalous roughening cannot occur in local growth models. However, some conserved dynamics may display superroughening if a given type of term is present. PMID- 15904250 TI - Quantum size effects in adatom island decay. AB - The decay of hexagonal Ag adatom islands on top of larger Ag adatom islands on a Ag(111) surface is followed by a fast-scanning tunneling microscope. Islands do not always show the expected increase in decay rate with decreasing island size. Rather, distinct quantum size effects are observed where the decay rate decreases significantly for islands with diameters of 6, 9.3, 12.6, and 15.6 nm. We show that electron confinement of the surface state electrons is responsible for this enhancement of the detachment barrier for adatoms from the island edge. PMID- 15904251 TI - Complex shape evolution of electromigration-driven single-layer islands. AB - The shape evolution of two-dimensional islands through periphery diffusion biased by an electromigration force is studied numerically using a continuum approach. We show that the introduction of crystal anisotropy in the mobility of edge atoms induces a rich variety of migration modes, which include oscillatory and irregular behavior. A phase diagram in the plane of anisotropy strength and island size is constructed. The oscillatory motion can be understood in terms of stable facets that develop on one side of the island, and which the island then slides past. The facet orientations are determined analytically. PMID- 15904252 TI - Generation and detection of shear acoustic waves in metal submicrometric films with ultrashort laser pulses. AB - We present experimental and calculational results demonstrating the thermoelastic generation of shear acoustic waves using femtosecond laser pulses in submicrometric isotropic aluminum films. We show that the generation of the shear waves is correlated to the reduction of the width of the optoacoustic source on the surface. The presence of shear waves is related to acoustic diffraction and acoustic mode conversion at the thin film interfaces. PMID- 15904253 TI - Geometry of crumpled paper. AB - We measure the geometry of a crumpled sheet of paper with laser-aided topography and discuss its statistical properties. The curvature of an elastoplastic fold scales linearly with applied force. The curvature distribution follows an exponential form with regions of high curvature localized along ridges. The measured ridge length distribution is consistent with a hierarchical model for ridge breaking during crumpling. A large fraction of the ridges are observed to terminate without bifurcating, and the ridge network connectedness is not as complete as anticipated. The self-affinity of the surface is characterized by a Hurst exponent of 0.71+/-0.01 in contrast with previous results. PMID- 15904254 TI - High-pressure ground state of SmB6: electronic conduction and long range magnetic order. AB - High-pressure 149Sm nuclear forward scattering of synchrotron radiation and specific heat measurements have been performed on the intermediate valent Kondo insulator SmB6. The results show that at a critical pressure p(c) approximately = 6 GPa, where the charge gap closes, a first order transition occurs to a magnetically ordered state, which shows typical features of trivalent samarium compounds. The similarity with SmS stresses the role of local correlations and gives important insight into the debate on the local or itinerant character of the f electrons in heavy fermion systems. PMID- 15904255 TI - Importance of interorbital charge transfers for the metal-to-insulator transition of BaVS3. AB - The underlying mechanism of the metal-to-insulator transition (MIT) in BaVS3 is investigated, using dynamical mean-field theory in combination with density functional theory. It is shown that correlation effects are responsible for a strong charge redistribution, which lowers the occupancy of the broader A(1g) band in favor of the narrower E(g) bands and thereby substantially modifies the Fermi surface. This resolves several discrepancies between band theory and the experimental findings, such as the observed value of the charge-density-wave ordering vector associated with the MIT, and the presence of local moments in the metallic phase. PMID- 15904256 TI - Metal-insulator-like behavior in semimetallic bismuth and graphite. AB - When high quality bismuth or graphite crystals are placed in a magnetic field directed along the c axis (trigonal axis for bismuth) and the temperature is lowered, the resistance increases as it does in an insulator but then saturates. We show that the combination of unusual features specific to semimetals, i.e., low carrier density, small effective mass, high purity, and an equal number of electrons and holes (compensation), gives rise to a unique ordering and spacing of three characteristic energy scales, which not only is specific to semimetals but which concomitantly provides a wide window for the observation of apparent field-induced metal-insulator behavior. Using magnetotransport and Hall measurements, the details of this unusual behavior are captured with a conventional multiband model, thus confirming the occupation by semimetals of a unique niche between conventional metals and semiconductors. PMID- 15904257 TI - Photoinduced charge currents in mesoscopic rings. AB - The temporal and spatial controllability of charge distribution in submicron structures opens new avenues for potential applications and for the understanding of nonequilibrium processes. Here we suggest a novel way to trigger and control within picoseconds charge currents and magnetic moments in nanoscopic and mesoscopic ring structures by applying two shaped, time-delayed light pulses. Our quantum dynamic calculations show that the magnitude and direction of the induced currents are tunable by varying the time delay and strengths of the pulses. Furthermore, in an array of rings desirable magnetic orders are generated depending on the ring sizes and particle number. PMID- 15904258 TI - Topologically protected qubits from a possible non-Abelian fractional quantum Hall state. AB - The Pfaffian state is an attractive candidate for the observed quantized Hall plateau at a Landau-level filling fraction nu=5/2. This is particularly intriguing because this state has unusual topological properties, including quasiparticle excitations with non-Abelian braiding statistics. In order to determine the nature of the nu=5/2 state, one must measure the quasiparticle braiding statistics. Here, we propose an experiment which can simultaneously determine the braiding statistics of quasiparticle excitations and, if they prove to be non-Abelian, produce a topologically protected qubit on which a logical Not operation is performed by quasiparticle braiding. Using the measured excitation gap at nu=5/2, we estimate the error rate to be 10(-30) or lower. PMID- 15904259 TI - Current fluctuations in an interacting quantum dot. AB - We calculate the counting statistics of electron transfer through an open quantum dot with charging interaction. A dot that is connected to leads by two single channel quantum point contacts in an in-plane magnetic field is described by a Luttinger liquid with impurity at the Toulouse point. We find that the fluctuations of the current through this conductor exhibit distinctive interaction effects. Fluctuations saturate at high voltages, while the mean current increases linearly with the bias voltage. All cumulants higher than the second one reach at large bias a temperature independent limit. PMID- 15904260 TI - Universality of the edge-tunneling exponent of fractional quantum Hall liquids. AB - In a microscopic model of fractional quantum Hall liquids with electron-electron interactions and confinement, we calculate the edge Green's function via exact diagonalization. Our results for nu=1/3 and 2/3 suggest that, in the presence of Coulomb interaction, "external" parameters such as the sharpness of the edge and the strength of the edge confining potential, which can lead to edge reconstruction, may cause deviations from universality in the edge-tunneling I-V exponent. In particular, we do not find any direct dependence of this exponent on the range of the interaction potential as suggested by recent calculations in contradiction to the topological nature of the edge. PMID- 15904261 TI - Initial decoherence in solid state qubits. AB - We study decoherence due to low frequency noise in Josephson qubits. Non Markovian classical noise due to switching impurities determines inhomogeneous broadening of the signal. The theory is extended to include effects of high frequency quantum noise, due to impurities or to the electromagnetic environment. The interplay of slow noise with intrinsically non-Gaussian noise sources may explain the rich physics observed in the spectroscopy and in the dynamics of charge based devices. PMID- 15904262 TI - Nanomechanical magnetization reversal. AB - The dynamics of the ferromagnetic order parameter in thin magnetic films is strongly affected by the magnetomechanical coupling at certain resonance frequencies. By solving the equation of motion of the coupled mechanical and magnetic degrees of freedom we show that the magnetic field induced magnetization switching can be strongly accelerated by the lattice and illustrate the possibility of magnetization reversal by mechanical actuation. PMID- 15904263 TI - Parametric generation of forward and phase-conjugated spin-wave bullets in magnetic films. AB - We show experimentally as well as by numerical simulation that interaction of a linear two-dimensional spin-wave packet with quasiuniform pulsed pumping leads to the formation of strongly self-focused nonlinear spin-wave bullets propagating in both forward and reversed directions. The focusing of the reversed, phase conjugated wave bullet is stronger than that of the forward one, because not only the nonlinear four-wave self-focusing effect but also linear focusing due to two dimensional phase conjugation contributes to the focusing of the reversed bullet. PMID- 15904264 TI - Ab initio theory of dynamical core-hole screening in graphite from x-ray absorption spectra. AB - We have implemented the effect of dynamical core-hole screening, as given by Mahan, Nozieres, and De Dominicis, in a first-principles based method and applied the theory to the x-ray absorption (XA) spectrum of graphite. It turns out that two of the conspicuous peaks of graphite are well described, both regarding the position, shape, and relative intensity, whereas one peak is absent in the theory. Only by incorporation of both excitonic and delocalized processes can a full account of the experimental spectrum be obtained theoretically, and we interpret the XA spectrum in graphite to be the result of a well screened and a poor screened process, much in the same way as is done for core level x-ray photoelectron spectroscopy. PMID- 15904265 TI - Hysteresis and competition between disorder and crystallization in sheared and vibrated granular flow. AB - Experiments on spherical particles in a 3D annular shear cell vibrated from below and sheared from above show a hysteretic freezing or melting transition. Under sufficient vibration a crystallized state is observed, which can be melted by sufficient shear. The critical line for this transition coincides with equal kinetic energies for vibration and shear. The force distribution is double peaked in the crystalline state and single peaked with an approximately exponential tail in the disordered state. Continuous relations between pressure and volume (with dP/dV>0) exist for a continuum of partially and/or intermittently melted states over a range of parameters. PMID- 15904266 TI - Scale-rich metabolic networks. AB - Recent research in biology has clarified many features of the global organization of metabolic networks, including the biochemical mechanisms responsible for power laws in metabolite degrees. The primary aim of this Letter is to give the simplest possible biochemical explanations and minimal toy models based on a highly optimized tolerance perspective, which show where and why metabolic networks have power laws. A second aim is to further explain why scale-free explanations fail in this case to correctly describe metabolism. PMID- 15904267 TI - Spiking neurons learning phase delays: how mammals may develop auditory time difference sensitivity. AB - Time differences between the two ears are an important cue for animals to azimuthally locate a sound source. The first binaural brainstem nucleus, in mammals the medial superior olive, is generally believed to perform the necessary computations. Its cells are sensitive to variations of interaural time differences of about 10 micros. The classical explanation of such a neuronal time difference tuning is based on the physical concept of delay lines. Recent data, however, are inconsistent with a temporal delay and rather favor a phase delay. By means of a biophysical model we show how spike-timing-dependent synaptic learning explains precise interplay of excitation and inhibition and, hence, accounts for a physical realization of a phase delay. PMID- 15904268 TI - Enumeration of RNA structures by matrix models. AB - We enumerate the number of RNA contact structures according to their genus, i.e., the topological character of their pseudoknots. By using a recently proposed matrix model formulation for the RNA folding problem, we obtain exact results for the simple case of an RNA molecule with an infinitely flexible backbone, in which any arbitrary pair of bases is allowed. We analyze the distribution of the genus of pseudoknots as a function of the total number of nucleotides along the phosphate-sugar backbone. PMID- 15904269 TI - Computational complexity and fundamental limitations to fermionic quantum Monte Carlo simulations. AB - Quantum Monte Carlo simulations, while being efficient for bosons, suffer from the "negative sign problem" when applied to fermions--causing an exponential increase of the computing time with the number of particles. A polynomial time solution to the sign problem is highly desired since it would provide an unbiased and numerically exact method to simulate correlated quantum systems. Here we show that such a solution is almost certainly unattainable by proving that the sign problem is nondeterministic polynomial (NP) hard, implying that a generic solution of the sign problem would also solve all problems in the complexity class NP in polynomial time. PMID- 15904270 TI - Fixed points of the dissipative Hofstadter model. AB - The phase diagram of a dissipative particle in a periodic potential and a magnetic field is studied in the weak barrier limit and in the tight binding regime. For the case of half flux per plaquette, and for a wide range of values of the dissipation, the physics of the model is determined by a nontrivial fixed point. A combination of exact and variational results is used to characterize this fixed point. Finally, it is also argued that there is an intermediate energy scale that separates the weak coupling physics from the tight binding solution. PMID- 15904271 TI - Dynamics of the BCS-BEC crossover in a degenerate Fermi gas. AB - We study the short-time dynamics of a degenerate Fermi gas positioned near a Feshbach resonance following an abrupt jump in the atomic interaction resulting from a change of magnetic field. We investigate the dynamics of the condensate order parameter and pair wave function for a range of field strengths. When the jump is sufficient to span the BCS to Bose-Einstein condensation crossover, we show that the rigidity of the momentum distribution precludes any atom-molecule oscillations in the entrance channel dominated resonances observed in 40K and 6Li. Focusing on material parameters tailored to the 40K Feshbach resonance at 202.1 G, we comment on the integrity of the fast sweep projection technique as a vehicle to explore the condensed phase in the crossover region. PMID- 15904272 TI - Photon recoil momentum in dispersive media. AB - A systematic shift of the photon recoil momentum due to the index of refraction of a dilute gas of atoms has been observed. The recoil frequency was determined with a two-pulse light grating interferometer using near-resonant laser light. The results show that the recoil momentum of atoms caused by the absorption of a photon is n variant Planck's k, where n is the index of refraction of the gas and k is the vacuum wave vector of the photon. This systematic effect must be accounted for in high-precision atom interferometry with light gratings. PMID- 15904273 TI - Damping of a unitary Fermi gas. AB - We measure the temperature dependence of the radial breathing mode in an optically trapped, unitary Fermi gas of 6Li, just above the center of a broad Feshbach resonance. The damping rate reveals a clear change in behavior which we interpret as arising from a superfluid transition. We suggest pair breaking as a mechanism for an increase in the damping rate which occurs at temperatures well above the transition. In contrast to the damping, the frequency varies smoothly and remains close to the unitary hydrodynamic value. At low temperature T, the damping depends on the atom number only through the reduced temperature, and extrapolates to 0 at T = 0. PMID- 15904274 TI - Origin of the usefulness of the natural-time representation of complex time series. AB - The concept of natural time turned out to be useful in revealing dynamical features behind complex time series including electrocardiograms, ionic current fluctuations of membrane channels, seismic electric signals, and seismic event correlation. However, the origin of this empirical usefulness is yet to be clarified. Here, it is shown that this time domain is in fact optimal for enhancing the signals in time-frequency space by employing the Wigner function and measuring its localization property. PMID- 15904275 TI - How to measure subdiffusion parameters. AB - We propose a method to measure the subdiffusion parameter alpha and subdiffusion coefficient Dalpha which are defined by means of the relation chi2 = 2Dalpha / Gamma(1+alpha)(t alpha), where chi2 denotes a mean square displacement of a random walker starting from x = 0 at the initial time t = 0. The method exploits a membrane system where a substance of interest is transported in a solvent from one vessel to another across a thin membrane which plays here only an auxiliary role. We experimentally study a diffusion of glucose and sucrose in a gel solvent, and we precisely determine the parameters alpha and Dalpha, using a fully analytic solution of the fractional subdiffusion equation. PMID- 15904276 TI - Gamma-ray Constraint on galactic positron production by MeV dark matter. AB - The Galactic positrons, as observed by their annihilation gamma-ray line at 0.511 MeV, are difficult to account for with astrophysical sources. It has been proposed that they are produced instead by dark matter annihilation or decay in the inner Galactic halo. To avoid other constraints, these processes are required to occur "invisibly," such that the eventual positron annihilation is the only detectable signal. However, electromagnetic radiative corrections to these processes inevitably produce real gamma rays ("internal bremsstrahlung"); this emission violates COMPTEL and EGRET constraints unless the dark matter mass is less than about 20 MeV. PMID- 15904277 TI - Supersymmetric dark matter and the extragalactic gamma ray background. AB - We trace the origin of the newly determined extragalactic gamma-ray background from EGRET data to an unresolved population of blazars and neutralino annihilation in cold dark matter halos. Using results of high-resolution simulations of cosmic structure formation, we calculate composite spectra and compare with the EGRET data. The resulting best-fit value for the neutralino mass is m(chi) = 515(+110)(-75) GeV (systematic errors approximately 30%). PMID- 15904278 TI - Can dark matter annihilation dominate the extragalactic gamma-ray background? AB - Annihilating dark matter (DM) has been discussed as a possible source of gamma rays from the galactic center and as a contribution to the extragalactic gamma ray background. Assuming universality of the density profile of DM halos, we show that it is quite unlikely that DM annihilation is a main constituent of extragalactic gamma-ray background, without exceeding the observed gamma-ray flux from the galactic center. This argument becomes stronger when we include enhancement of the density profiles by supermassive black holes or baryon cooling. The presence of a substructure may loosen the constraint, but only if a very large cross section as well as the rather flat profile are realized. PMID- 15904281 TI - Mass of the Bc meson in three-flavor lattice QCD. AB - We use lattice QCD to predict the mass of the Bc meson. We use the MILC Collaboration's ensembles of lattice gauge fields, which have a quark sea with two flavors much lighter than a third. Our final result is mBc = 6304+/-12(+18)( 0) MeV. The first error bar is a sum in quadrature of statistical and systematic uncertainties, and the second is an estimate of heavy-quark discretization effects. PMID- 15904282 TI - The hot nonperturbative gluon plasma is an almost ideal colored liquid. AB - We study properties of a gluon plasma above the critical temperature Tc in a generalized quasiparticle approach with a Lorentz spectral function. The model parameters are determined by a fit of the entropy s to lattice QCD data. The effective degrees of freedom are found to be rather heavy and of a sizable width. With the spectral width being closely related to the interaction rate, we find a large effective cross section, which is comparable to the typical distance squared of the quasiparticles. This suggests that the system should be viewed as a liquid as also indicated by an estimate of the plasma parameter Gamma. Furthermore, within the quasiparticle approach we find a very low viscosity to entropy ratio, eta/s approximately 0.2 for T > 1.05 Tc, supporting the recent conjecture of an almost ideal quark-gluon liquid seen at RHIC. PMID- 15904284 TI - Dipole-dipole excitation and ionization in an ultracold gas of Rydberg atoms. AB - In cold dense Rydberg atom samples, the dipole-dipole interaction strength is effectively resonant at the typical interatomic spacing in the sample, and the interaction has a 1/R3 dependence on interatomic spacing R. The dipole-dipole attraction leads to ionizing collisions of initially stationary atoms, which produces hot atoms and ions and initiates the evolution of initially cold samples of neutral Rydberg atoms into plasmas. More generally, the strong dipole-dipole forces lead to motion, which must be considered in proposed applications. PMID- 15904283 TI - "Safe" Coulomb excitation of 30Mg. AB - We report on the first radioactive beam experiment performed at the recently commissioned REX-ISOLDE facility at CERN in conjunction with the highly efficient gamma spectrometer MINIBALL. Using 30Mg ions accelerated to an energy of 2.25 MeV/u together with a thin (nat)Ni target, Coulomb excitation of the first excited 2+ states of the projectile and target nuclei well below the Coulomb barrier was observed. From the measured relative deexcitation gamma-ray yields the B(E2;0(+)gs-->2(+)1) value of 30Mg was determined to be 241(31)e2 fm4. Our result is lower than values obtained at projectile fragmentation facilities using the intermediate-energy Coulomb excitation method, and confirms the theoretical conjecture that the neutron-rich magnesium isotope 30Mg resides outside the "island of inversion." PMID- 15904285 TI - High-accuracy optical clock via three-level coherence in neutral bosonic 88Sr. AB - An optical atomic clock scheme is proposed that utilizes two lasers to establish coherent coupling between the 5s2 1S0 ground state of 88Sr and the first excited state, 5s5p 3P0. The coupling is mediated by the broad 5s5p 1P1 state, exploiting the phenomenon of electromagnetically induced transparency. The effective linewidth of the clock transition can be chosen at will by adjusting the laser intensity. By trapping the 88Sr atoms in an optical lattice, long interaction times with the two lasers are ensured; Doppler and recoil effects are eliminated. Based on a careful analysis of systematic errors, a clock accuracy of better than 2 x 10(-17) is expected. PMID- 15904286 TI - Electron collisions with laser cooled and trapped metastable helium atoms: total scattering cross sections. AB - Absolute measurements of total scattering cross sections for low energy (5-70 eV) electrons by metastable helium (2(3)S) atoms are presented. The measurements are performed using a magneto-optical trap which is loaded from a laser-cooled, bright beam of slow He(2(3)S) atoms. The data are compared with predictions from convergent close coupling and R matrix with pseudostate calculations, and we find good agreement between experiment and theory. PMID- 15904280 TI - Measurement of charged-particle multiplicities in gluon and quark jets in pp collisions at square root of s = 1.8 TeV. AB - We report the first largely model independent measurement of charged particle multiplicities in quark and gluon jets, Nq and Ng, produced at the Fermilab Tevatron in pp collisions with a center-of-mass energy of 1.8 TeV and recorded by the Collider Detector at Fermilab. The measurements are made for jets with average energies of 41 and 53 GeV by counting charged particle tracks in cones with opening angles of theta(c) = 0.28, 0.36, and 0.47 rad around the jet axis. The corresponding jet hardness Q = Ejet theta c varies in the range from 12 to 25 GeV. At Q = 19.2 GeV, the ratio of multiplicities r = Ng/Nq is found to be 1.64+/ 0.17, where statistical and systematic uncertainties are added in quadrature. The results are in agreement with resummed perturbative QCD calculations. PMID- 15904287 TI - Melting, premelting, and structural transitions in size-selected aluminum clusters with around 55 atoms. AB - Heat capacities have been determined for unsupported aluminum clusters, Al49(+) - Al63(+), from 150 to 1050 K. Peaks in the heat capacities due to melting occur between 450 and 650 K (well below the bulk melting point of 933 K). The peaks for Al+51 and Al+52 are bimodal, suggesting the presence of a premelting transition where the surface of the clusters melts around 100 K before the core. For clusters with n > 55 the melting temperatures suddenly drop, and there is a dip in the heat capacities due to a transition between two solid forms before the clusters melt. PMID- 15904288 TI - Experimental observation of classical subwavelength interference with a pseudothermal light source. AB - We report the experimental observation of classical subwavelength double slit interference with a pseudothermal light source. The experimental results are in good agreement with the theoretical simulation using the second order correlation function for the thermal light. PMID- 15904289 TI - Computable measure of nonclassicality for light. AB - We propose the entanglement potential (EP) as a measure of nonclassicality for quantum states of a single-mode electromagnetic field. It is the amount of two mode entanglement that can be generated from the field using linear optics, auxiliary classical states, and ideal photodetectors. The EP detects nonclassicality, has a direct physical interpretation, and can be computed efficiently. These three properties together make it stand out from previously proposed nonclassicality measures. We derive closed expressions for the EP of important classes of states and analyze as an example of the degradation of nonclassicality in lossy channels. PMID- 15904290 TI - Complete destructive interference of partially coherent sources of acoustic waves. AB - Theoretical analysis has recently shown that the optical fields from several point sources may exhibit complete destructive interference even if the sources are not fully coherent with respect to each other. The experimental verification of this statement in the optical domain is not easy. In this Letter we demonstrate the effect using acoustical waves instead of light waves. PMID- 15904279 TI - Search for factorization-suppressed B-->chi(c)K(*) decays. AB - We search for the factorization-suppressed decays B-->chi(c0)K(*) and B- >chi(c2)K(*), with chi(c0) and chi(c2) decaying into J/psi gamma, using a sample of 124 x 10(6) BB events collected with the BABAR detector at the PEP-II storage ring of the Stanford Linear Accelerator Center. We find no significant signal and set upper bounds for the branching fractions. PMID- 15904291 TI - Frequency bistability of a semiconductor laser under a frequency-dependent feedback. AB - The emission frequency of a diode laser submitted to a frequency-dependent optoelectronic feedback is observed to have more than one stable operation point together with a stable power emission. This is, to our knowledge, the first observation of bistability exclusively in the frequency of an optical system. The experiment was carried out with a semiconductor laser coupled to the cesium D2 line by an orthogonally polarized frequency-sensitive optical feedback. PMID- 15904292 TI - High harmonic XUV spectral phase interferometry for direct electric-field reconstruction. AB - We demonstrate the first experimental complete temporal characterization of high harmonic XUV pulses by spectral phase interferometry, with an all-optical setup. This method allows us to perform single-shot measurements of the harmonic temporal profile and phase, revealing a remarkable shot-to-shot stability. We characterize harmonics generated in argon by a 50 fs 800 nm laser pulse. The 11th harmonic is found to be 22 fs long with a negative chirp rate of -4.8 x 10(27) s( 2). This duration can be reduced to 13 fs by modulating the polarization of the generating laser. The technique is easy to implement and could be routinely used in femtosecond XUV pump-probe experiments with harmonics. PMID- 15904293 TI - Coherent plasmonic enhanced terahertz transmission through random metallic media. AB - We experimentally show coherent, enhanced terahertz transmission through dense 3D random metallic media having subwavelength heterogeneity. Preservation of the incident polarization state and strong dispersion of the transmitted radiation indicate that the enhanced transmission is due to delocalized plasmonic propagation over distances 5 orders of magnitude greater than the skin depth. The experimental observations are supported by numerical finite difference time domain simulations. PMID- 15904294 TI - Spontaneous optical fractal pattern formation. AB - We report, for the first time, spontaneous nonlinear optical spatial fractals. The proposed generic mechanism employs intrinsic nonlinear dynamics both to generate an initial pattern seed and to fill out structure across decades of spatial scale. We demonstrate this in one of the simplest of nonlinear optical systems, composed of a Kerr slice and a single-feedback mirror. In this case, the smallest pattern scales are limited by either the optical wavelength or the diffusion length of the medium photoexcitation. The dimension characteristics of these particular fractals are also derived. PMID- 15904295 TI - Phase compactons in chains of dispersively coupled oscillators. AB - We study the phase dynamics of a chain of autonomous oscillators with a dispersive coupling. In the quasicontinuum limit the basic discrete model reduces to a Korteveg-de Vries-like equation, but with a nonlinear dispersion. The system supports compactons: solitary waves with a compact support and kovatons which are compact formations of glued together kink-antikink pairs that may assume an arbitrary width. These robust objects seem to collide elastically and, together with wave trains, are the building blocks of the dynamics for typical initial conditions. Numerical studies of the complex Ginzburg-Landau and Van der Pol lattices show that the presence of a nondispersive coupling does not affect kovatons, but causes a damping and deceleration or growth and acceleration of compactons. PMID- 15904296 TI - Ballistic and localized transport for the atom optics kicked rotor in the limit of a vanishing kicking period. AB - We present mean energy measurements for the atom optics kicked rotor as the kicking period tends to zero. A narrow resonance is observed marked by quadratic energy growth, in parallel with a complete freezing of the energy absorption away from the resonance peak. Both phenomena are explained by classical means, taking proper account of the atoms' initial momentum distribution. PMID- 15904297 TI - Transitions to electrochemical turbulence. AB - We report experimental evidence of transitions from limit cycle oscillations through a phase turbulent regime to space-time defect turbulence in a spatially (quasi-)one-dimensional electrochemical system with nonlocal coupling. The transitions are characterized in terms of the defect density, the Karhunen-Loeve decomposition dimension, and a measure of the degree of spatial correlation in the data. Furthermore, these quantities give the first experimental confirmation that the spatial coupling range in electrochemical systems indeed depends on the distance between the working and the counterelectrode. PMID- 15904298 TI - Second mode of recycling together with period doubling links single-bubble and multibubble sonoluminescence. AB - We report the existence of a second type of recycling mode that occurs for air seeded bubbles. Observation of period doubling in both the stable, the first type, and the second type of recycling mode, together with simultaneous measurement of the relative phase of light emission compared to the drive, shows that the instability boundaries of period doubling and bubble extinction are mainly determined by the bubble size irregardless of the gas composition. The second type mode seems to represent a link between single-bubble and multibubble sonoluminescence. PMID- 15904299 TI - Theoretical and numerical approach to "magic angle" of stone skipping. AB - We investigate the condition for the bounce of circular disks which obliquely impacts on the fluid surface. An experiment [C. Clanet, F. Hersen, and L. Bocquet, Nature (London) 427, 29 (2004)] revealed that there exists a "magic angle" of 20 degrees between a disk's face and water surface in which the condition of the lowest impact speed necessary for a bounce is minimized. We perform a three-dimensional simulation of the disk-water impact by means of the smoothed particle hydrodynamics. Furthermore, we analyze the impact with a model of the ordinary differential equation (ODE). Our simulation is in good agreement with the experiment. The analysis with the ODE model gives us a theoretical insight into the "magic angle" of stone skipping. PMID- 15904300 TI - Drag reduction by microbubbles in turbulent flows: the limit of minute bubbles. AB - Drag reduction by microbubbles is a promising engineering method for improving ship performance. A fundamental theory of the phenomenon is lacking, however, making actual design quite haphazard. We offer here a theory of drag reduction by microbubbles in the limit of very small bubbles, when the effect of the bubbles is mainly to normalize the density and the viscosity of the carrier fluid. The theory culminates with a prediction of the degree of drag reduction given the concentration profile of the bubbles. Comparisons with experiments are discussed and the road ahead is sketched. PMID- 15904301 TI - Density fluctuations in strongly stratified two-dimensional turbulence. AB - We present a study of density fluctuations in two-dimensional soap-film convection. When the temperature difference (DeltaT) imposed vertically along the film is smaller than a critical value (DeltaT(C) approximately 48 K), the convective motion is in a strongly stratified state and the frequency power spectrum of density fluctuations shows a Bolgiano-like scaling f(-7/5) in the buoyancy subrange. When DeltaT>DeltaT(C), the fluid motion crosses over to a strongly mixed state characterized by the emergence of a large-scale circulation. The density power spectrum in this state has a passive-scalar-like scaling f( 1.0). PMID- 15904302 TI - X-ray resonance in crystal cavities: realization of Fabry-Perot resonator for hard x rays. AB - X-ray back diffraction from monolithic two silicon crystal plates of 25-150 microm thickness and a 40-150 microm gap using synchrotron radiation of energy resolution DeltaE = 0.36 meV at 14.4388 keV clearly show resonance fringes inside the energy gap and the total-reflection range for the (12 4 0) reflection. This cavity resonance results from the coherent interaction between the x-ray wave fields generated by the two plates with a gap smaller than the x-ray coherence length. This finding opens up new opportunities for high-resolution and phase contrast x-ray studies, and may lead to new developments in x-ray optics. PMID- 15904303 TI - Mechanism of stabilization of ballooning modes by toroidal rotation shear in tokamaks. AB - A ballooning perturbation in a toroidally rotating tokamak is expanded by square integrable eigenfunctions of an eigenvalue problem associated with ballooning modes in a static plasma. Especially a weight function is chosen such that the eigenvalue problem has only the discrete spectrum. The eigenvalues evolve in time owing to toroidal rotation shear, resulting in a countably infinite number of crossings among them. The crossings cause energy transfer from an unstable mode to the infinite number of stable modes; such transfer works as the stabilization mechanism of the ballooning mode. PMID- 15904304 TI - Observation of centrifugally driven interchange instabilities in a plasma confined by a magnetic dipole. AB - Centrifugally driven interchange instabilities are observed in a laboratory plasma confined by a dipole magnetic field. The instabilities appear when an equatorial mesh is biased to drive a radial current that causes rapid axisymmetric plasma rotation. The observed instabilities are quasicoherent in the laboratory frame of reference; they have global radial mode structures and low azimuthal mode numbers, and they are modified by the presence of energetic, magnetically confined electrons. Results from a self-consistent nonlinear simulation reproduce the measured mode structures. PMID- 15904305 TI - Observation of a transition from fluid to kinetic nonlinearities for langmuir waves driven by stimulated Raman backscatter. AB - Thomson scattering is used to measure Langmuir waves (LW) driven by stimulated Raman scattering (SRS) in a diffraction limited laser focal spot. For SRS at wave numbers klambda(D) less similar 0.29, where k is the LW number and lambda(D) is the Debye length, multiple waves are detected and are attributed to the Langmuir decay instability (LDI) driven by the primary LW. At klambda(D) greater similar 0.29, a single wave, frequency-broadened spectrum is observed. The transition from the fluid to the kinetic regime is qualitatively consistent with particle-in cell simulations and crossing of the LDI amplitude threshold above that for LW self-focusing. PMID- 15904306 TI - Exact energy principle in magnetic reconnection for current-sheet models. AB - On the basis of an exact nonlinear energy principle, it is shown that the change in magnetic topology (i.e., reconnection) in a finite-domain system leads to the conversion of magnetic field energy to particle energy. However, it is also shown that the conversion efficiency gradually disappears as the system size increases. This principle is demonstrated with model current-sheet equilibria including Harris and Fadeev solutions, as well as a current-sheet equilibrium which contains a singular current layer. The finding that energy conversion in reconnection is highly dependent on the system size may have an important implication for numerical simulations performed under finite geometry. PMID- 15904307 TI - Modified jeans instability for dust grains in a plasma. AB - An investigation of the properties of linear stability is conducted for a system consisting of particles having mass m and charge q, interacting through the gravitational and electrostatic force (Jeans instability). However, in light of recent works showing that dust particles in a plasma can have a Lennard-Jones like shielding potential, a new set of equations has been derived, where the electrostatic interaction among the dust particles is Lennard-Jones-like instead of Coulomb-like. A new condition for the gravitational instability is derived, showing a broader spectrum of unstable modes with faster growth rates. PMID- 15904308 TI - Effective lagrangian and topological interactions in supersolids. AB - We construct a low-energy effective Lagrangian describing zero temperature supersolids. Galilean invariance imposes strict constraints on the form of the effective Lagrangian. We identify a topological term in the Lagrangian that couples superfluid and crystalline modes. For small superfluid fractions, this interaction term is dominant in problems involving defects. As an illustration, we compute the differential cross section of scatterings of low-energy transverse elastic phonons by a superfluid vortex. The result is model independent. PMID- 15904309 TI - Titanium-decorated carbon nanotubes as a potential high-capacity hydrogen storage medium. AB - We report a first-principles study, which demonstrates that a single Ti atom coated on a single-walled nanotube (SWNT) binds up to four hydrogen molecules. The first H2 adsorption is dissociative with no energy barrier while the other three adsorptions are molecular with significantly elongated H-H bonds. At high Ti coverage we show that a SWNT can strongly adsorb up to 8 wt % hydrogen. These results advance our fundamental understanding of dissociative adsorption of hydrogen in nanostructures and suggest new routes to better storage and catalyst materials. PMID- 15904310 TI - Radial elasticity of multiwalled carbon nanotubes. AB - We report an experimental and a theoretical study of the radial elasticity of multiwalled carbon nanotubes as a function of external radius. We use atomic force microscopy and apply small indentation amplitudes in order to stay in the linear elasticity regime. The number of layers for a given tube radius is inferred from transmission electron microscopy, revealing constant ratios of external to internal radii. This enables a comparison with molecular dynamics results, which also shed some light onto the applicability of Hertz theory in this context. Using this theory, we find a radial Young modulus strongly decreasing with increasing radius and reaching an asymptotic value of 30+/-10 GPa. PMID- 15904311 TI - Dynamic model of super-arrhenius relaxation rates in glassy materials. AB - Super-Arrhenius relaxation rates in glassy materials can be associated with thermally activated rearrangements of increasing numbers of molecules at decreasing temperatures. We explore a model of such a mechanism in which stringlike fluctuations in the neighborhood of shear transformation zones provide routes along which rearrangements can propagate, and the entropy associated with critically long strings allows the rearrangement to be distributed stably in the surrounding material. We further postulate that, at low enough temperatures, these fluctuations are localized on the interfaces between frustration-limited domains, and in this way obtain a modified Vogel-Fulcher formula for the relaxation rate. PMID- 15904312 TI - Superheating and induced melting at semiconductor interfaces. AB - We present ab initio density-functional simulations of the state of several semiconductor surfaces at temperatures near the bulk melting temperatures. We find that the solid-liquid phase-transition temperature at the surface can be altered via a microscopic (single-monolayer) coating with a different lattice matched semiconducting material. Our results show that a single-monolayer GaAs coating on a Ge(110) surface above the Ge melting temperature can dramatically reduce the diffusion coefficient of the germanium atoms, going so far as to prevent melting of the bulk layers on the 10 ps time scale. In contrast, a single monolayer coating of Ge on a GaAs(110) surface introduces defects into the bulk and induces melting of the top layer of GaAs atoms 300 K below the GaAs melting point. To our knowledge, these calculations represent the first ab initio investigation of the superheating and induced melting phenomena. PMID- 15904313 TI - Strain phase diagram and domain orientation in SrTiO3 thin films. AB - SrTiO3 thin films were used as a model system to study the effects of strain and epitaxial constraint on structural phase transitions of perovskite films. The basic phenomena revealed will apply to a variety of important structural transitions including the ferroelectric transition. Highly strained SrTiO3 films were grown on different substrates, providing both compressive and tensile strain. The measured strain-temperature phase diagram is qualitatively consistent with theory; however, the increase in the phase transition temperature is much larger than predicted. Because of the epitaxial strain and substrate clamping, the SrTiO3 lattice is tetragonal at all temperatures. The phase transitions involve only changes in internal symmetry. The low temperature phase under tensile strain has a unique structure with orthorhombic Cmcm space group but a tetragonal lattice, an interesting consequence of epitaxial constraint. PMID- 15904314 TI - Role of strain-dependent surface energies in Ge/Si(100) island formation. AB - Formation energies for Ge/Si(100) pyramidal islands are computed combining continuum calculations of strain energy with first-principles-computed strain dependent surface energies. The strain dependence of surface energy is critically impacted by the presence of strain-induced changes in the Ge {100} surface reconstruction. The appreciable strain dependencies of rebonded-step {105} and dimer-vacancy-line-reconstructed {100} surface energies are estimated to give rise to a significant reduction in the surface contribution to island formation energies. PMID- 15904315 TI - Towards quantitative understanding of formation and stability of Ge hut islands on Si(001). AB - We analyze Ge hut island formation on Si(001), using first-principles calculations of energies, stresses, and their strain dependence of Ge/Si(105) and Ge/Si(001) surfaces combined with continuum modeling. We give a quantitative assessment on strain stabilization of Ge(105) facets, estimate the critical size for hut nucleation or formation, and evaluate the magnitude of surface stress discontinuity at the island's edge and its effect on island stability. PMID- 15904316 TI - Experimental and theoretical investigation of single Cu, Ag, and Au atoms adsorbed on Si(111)-(7x7). AB - Using scanning tunneling microscopy and first principles calculations, the adsorption sites of single Cu, Ag, and Au atoms on the Si(111)-(7x7) surface have been systematically investigated and identified. Despite their monovalence, these atoms were found to adsorb at high coordination sites, seeking to saturate the maximum number of dangling bonds. The stable adsorption sites were further observed to be distinctly different in the faulted and unfaulted half unit cells, showing an asymmetry that has never been observed for many other adsorbates. PMID- 15904317 TI - Why are alkali halide surfaces not wetted by their own melt? AB - Alkali halide (100) crystal surfaces are anomalous, being very poorly wetted by their own melt at the triple point. We present extensive simulations for NaCl, followed by calculations of the solid-vapor, solid-liquid, and liquid-vapor free energies showing that solid NaCl(100) is a nonmelting surface, and that its full behavior can quantitatively be accounted for within a simple Born-Meyer-Huggins Fumi-Tosi model potential. The incomplete wetting is traced to the conspiracy of three factors: surface anharmonicities stabilizing the solid surface; a large density jump causing bad liquid-solid adhesion; incipient NaCl molecular correlations destabilizing the liquid surface. The latter is pursued in detail, and it is shown that surface short-range charge order acts to raise the surface tension because incipient NaCl molecular formation anomalously reduces the surface entropy of liquid NaCl much below that of solid NaCl(100). PMID- 15904318 TI - Observation of two-dimensional classical wave localization: third sound on superfluid 4He films on a disordered substrate. AB - We present the results of measurements of the propagation of third sound waves on superfluid 4He adsorbed to two-dimensional ordered and disordered substrates. In the disordered case we compare the experimental results to theoretical predictions of classical wave localization in such systems and conclude that classical wave localization is present in our system. PMID- 15904319 TI - Spin-dependent resistivity at transitions between integer quantum hall states. AB - The longitudinal resistivity at transitions between integer quantum Hall states in two-dimensional electrons confined to AlAs quantum wells is found to depend on the spin orientation of the partially filled Landau level in which the Fermi energy resides. The resistivity can be enhanced by an order of magnitude as the spin orientation of this energy level is aligned with the majority spin. We discuss possible causes and suggest a new explanation for the spikelike features observed at the edges of quantum Hall minima near Landau level crossings. PMID- 15904320 TI - Phase-space dynamics of semiclassical spin- 1/2 Bloch electrons. AB - Following recent interest in a kinetic description of the semiclassical Bloch electron dynamics, we propose a new formulation based on the previously developed Lie-Poisson formulation of dynamics. It includes modifications required to account for the Berry curvature contribution to the electron's equation of motion as well as essential ingredients of a quantum treatment of spin- 1/2 degrees of freedom. Our theory is also manifestly gauge invariant and thus permits inclusion of the electron interactions. The scope of our formulation extends beyond its solid state physics motivation and includes recently discussed noncommutative generalizations of classical mechanics as well as historically important models from quantum gravity physics. PMID- 15904321 TI - Pattern formation as a signature of quantum degeneracy in a cold exciton system. AB - The development of a Turing instability to a spatially modulated state in a photoexcited electron-hole system is proposed as a novel signature of exciton Bose statistics. We show that such an instability, driven by kinetics of exciton formation, can result from stimulated processes that build up near quantum degeneracy. The stability of an electron-hole interface which describes recently observed exciton rings is analyzed. Interface instability occurs below a critical temperature, with a periodic 1D pattern developing via a continuous (type II) transition, in a qualitative agreement with observations. PMID- 15904322 TI - Temperature-dependent asymmetry of the nonlocal spin-injection resistance: evidence for spin nonconserving interface scattering. AB - We report nonlocal spin injection and detection experiments on mesoscopic Co Al2O3-Cu spin valves. We have observed a temperature-dependent asymmetry in the nonlocal resistance between parallel and antiparallel configurations of the magnetic injector and detector. This strongly supports the existence of a nonequilibrium resistance that depends on the relative orientation of the detector magnetization and the nonequilibrium magnetization in the normal metal providing evidence for increasing interface spin scattering with temperature. PMID- 15904323 TI - Vibrational sidebands and the Kondo effect in molecular transistors. AB - Electron transport through molecular quantum dots coupled to a single vibrational mode is studied in the Kondo regime. We apply a generalized Schrieffer-Wolff transformation to determine the effective low-energy spin-spin-vibron interaction. From this model we calculate the nonlinear conductance and find Kondo sidebands located at bias voltages equal to multiples of the vibron frequency. Because of selection rules, the side peaks are found to have strong gate-voltage dependences, which can be tested experimentally. In the limit of weak electron-vibron coupling, we employ a perturbative renormalization group scheme to calculate analytically the nonlinear conductance. PMID- 15904324 TI - Charge and spin dynamics of interacting fermions in a one-dimensional harmonic trap. AB - We study an atomic Fermi gas interacting through repulsive contact forces in a one-dimensional harmonic trap. Bethe-ansatz solutions lead to an inhomogeneous Tomonaga-Luttinger model for the low energy excitations. The equations of motion for charge and spin density waves are analyzed both near the trap center and near the trap edges. While the center shows conventional spin-charge separation, the edges cause a giant increase of the separation between these modes. PMID- 15904325 TI - Electric field modulation of galvanomagnetic properties of mesoscopic graphite. AB - Electric field effect devices based on mesoscopic graphite are fabricated for galvanomagnetic measurements. Strong modulation of magnetoresistance and Hall resistance as a function of the gate voltage is observed as the sample thickness approaches the screening length. Electric field dependent Landau level formation is detected from Shubnikov-de Haas oscillations. The effective mass of electron and hole carriers has been measured from the temperature dependent behavior of these oscillations. PMID- 15904326 TI - Vortices, tunneling, and deconfinement in bilayer quantum Hall excitonic superfluid. AB - The physics of vortices, instantons, and deconfinement is studied for layered superfluids in connection to bilayer quantum Hall systems at filling fraction nu = 1. We develop an effective gauge theory taking into account both vortices and instantons induced by interlayer tunneling. The renormalization group flow of the gauge charge and the instanton fugacity shows that the coupling of the gauge field to vortex matter produces a continuous transition between the confining phase of free instantons and condensed vortices and a deconfined gapless superfluid where magnetic charges are bound into dipoles. The interlayer tunneling conductance and the layer-imbalance induced inhomogeneous exciton condensate are discussed in connection to experiments. PMID- 15904327 TI - Wedge-shaped potential and Airy-function electron localization in oxide superlattices. AB - Oxide superlattices and microstructures hold the promise for creating a new class of devices with unprecedented functionalities. Density-functional studies of the recently fabricated, lattice-matched perovskite titanates (SrTiO3)n/(LaTiO3)m reveal a classic wedge-shaped potential well for the monolayer (m = 1) structure, originating from the Coulomb potential of a two-dimensional charged La sheet. The potential in turn confines the electrons in the Airy-function-localized states. Magnetism is suppressed for the monolayer structure, while in structures with a thicker LaTiO3 part, bulk antiferromagnetism is recovered, with a narrow transition region separating the magnetic LaTiO3 and the nonmagnetic SrTiO3. PMID- 15904328 TI - Bimolecular recombination coefficient as a sensitive testing parameter for low mobility solar-cell materials. AB - Bimolecular charge carrier recombination has been clarified in bulk heterojunction solar cells based on a blend of regioregular poly(3 hexylthiophene) and 1-(3-methoxycarbonyl)propyl-1-phenyl-[6,6]-methanofullerene using the time-of-flight method. We show how bimolecular recombination influences the charge carrier transport, how it limits the efficiency of low-mobility solar cells, and how to estimate the bimolecular recombination coefficient. We found that bimolecular recombination in these efficient photovoltaic materials is orders of magnitude slower as compared to Langevin recombination expected for low mobility materials. This effect is inherent to the nanomorphology of the bicontinuous interpenetrating network creating separate pathways for electrons and holes, and paves the way for the fabrication of bulk-heterojunction solar cells where bimolecular recombination is not the limiting factor. PMID- 15904329 TI - Vortex motion rectification in Josephson junction arrays with a ratchet potential. AB - By means of electrical transport measurements we have studied the rectified motion of vortices in ratchet potentials engineered on overdamped Josephson junction arrays. The rectified voltage as a function of the vortex density shows a maximum efficiency close a matching condition to the period of the ratchet potential indicating a collective vortex motion. Vortex current reversals were detected varying the driving force and vortex density revealing the influence of vortex-vortex interaction in the ratchet effect. PMID- 15904330 TI - Inhomogeneous level splitting in Pr 2-x BixRu2O7. AB - We report that Bi doping drives Pr 2-x BixRu2O7 from an antiferromagnetic insulator (x = 0) to a metallic paramagnet (x approximately 1) with a broad low T maximum in C/T. Neutron scattering reveals local low energy spin excitations (variant Planck's omega approximately 1 meV) with a spectrum that is unaffected by heating to k(B)T >> variant Planck's omega. We show that a continuous distribution of splittings of the non-Kramers Pr3+ ground-state doublet such as might result from various types of lattice strain can account for all the data. PMID- 15904331 TI - Spin excitations in an anisotropic bond-alternating quantum s = 1 chain in a magnetic field: contrast to haldane spin chains. AB - Inelastic neutron scattering experiments on the S = 1 quasi-one-dimensional bond alternating antiferromagnet Ni(C9D24N4)(NO2)ClO4 have been performed under magnetic fields below and above a critical field Hc at which the energy gap closes. Normal field dependence of Zeeman splitting of the excited triplet modes below Hc has been observed, but the highest mode is unusually small and smears out with increasing field. This can be explained by an interaction with a low lying two magnon continuum at q(parallel) = pi that is present in dimerized chains but absent in uniform ones. Above Hc, we find only one excited mode, in stark contrast with three massive excitations previously observed in the structurally similar Haldane-gap material NDMAP [A. Zheludev, Phys. Rev. B 68, 134438 (2003)]. PMID- 15904332 TI - Orbital correlations in the pseudocubic O and rhombohedral R phases of LaMnO3. AB - The local and intermediate structure of stoichiometric LaMnO3 has been studied in the pseudocubic and rhombohedral phases at high temperatures (300-1150 K). Neutron powder diffraction data were collected and a combined Rietveld and high real space resolution atomic pair distribution function analysis was carried out. The nature of the Jahn-Teller (JT) transition around 750 K is confirmed to be orbital order to disorder. In the high-temperature orthorhombic (O) and rhombohedral (R) phases, the MnO6 octahedra are still fully distorted locally. More importantly, the intermediate structure suggests the presence of local ordered clusters of diameter approximately 16 A ( approximately 4 MnO6 octahedra) implying strong nearest-neighbor JT antiferrodistortive coupling. These clusters persist well above the JT transition temperature even into the high-temperature R phase. PMID- 15904333 TI - Stimulated emission of surface plasmons at the interface between a silver film and an optically pumped dye solution. AB - Surface plasmons (SPs) are surface-bound electromagnetic waves supported by metals, offering the possibility of strong spatial confinement of electromagnetic fields on the micro- and nanoscales. They suffer, however, from strong damping caused by internal absorption and radiation losses. Here we demonstrate amplification of SPs by stimulated emission, which marks a possible solution to this problem. We use an attenuated-total-reflection setup to detect stimulated emission of SPs at the interface between a silver film and an optically pumped dye solution acting as the amplifying medium. Clear evidence of stimulated emission is provided by an excellent agreement of the experimental observations with a theoretical analysis. Amplification of SPs can be considered analogous to photon amplification in a laser, thereby suggesting novel approaches in the field of nano-optics. PMID- 15904334 TI - From bouncing to floating: noncoalescence of drops on a fluid bath. AB - When a drop of a viscous fluid is deposited on a bath of the same fluid, it is shown that its coalescence with this substrate is inhibited if the system oscillates vertically. Small drops lift off when the peak acceleration of the surface is larger than g. This leads to a steady regime where a drop can be kept bouncing for any length of time. It is possible to inject more fluid into the drop to increase its diameter up to several centimeters. Such a drop remains at the surface, forming a large sunk hemisphere. When the oscillation is stopped, the two fluids remain separated by a very thin air film, which drains very slowly (approximately 30 min). An analysis using lubrication theory accounts for most of the observations. PMID- 15904335 TI - Force indeterminacy in the jammed state of hard disks. AB - Granular packings of hard disks are investigated by means of contact dynamics which is an appropriate technique to explore the allowed force realizations in the space of contact forces. Configurations are generated for given friction coefficients, and then an ensemble of equilibrium forces is found for fixed contacts. We study the force fluctuations within this ensemble. In the limit of zero friction, the fluctuations vanish in accordance with the isostaticity of the packing. The magnitude of the fluctuations has a nonmonotonous friction dependence. The increase for small friction can be attributed to the opening of the angle of the Coulomb cone, while the decrease as friction increases is due to the reduction of connectivity of the contact network, leading to local, independent clusters of indeterminacy. We discuss the relevance of indeterminacy to packings of deformable particles and to the mechanical response properties. PMID- 15904336 TI - How hertzian solitary waves interact with boundaries in a 1D granular medium. AB - We perform measurements, numerical simulations, and quantitative comparisons with available theory on solitary wave propagation in a linear chain of beads without static preconstraint. By designing a nonintrusive force sensor to measure the impulse as it propagates along the chain, we study the solitary wave reflection at a wall. We show that the main features of solitary wave reflection depend on wall mechanical properties. Since previous studies on solitary waves have been performed at walls without these considerations, our experiment provides a more reliable tool to characterize solitary wave propagation. We find, for the first time, precise quantitative agreements. PMID- 15904337 TI - Exons, introns, and DNA thermodynamics. AB - The genes of eukaryotes are characterized by protein coding fragments, the exons, interrupted by introns, i.e., stretches of DNA which do not carry useful information for protein synthesis. We have analyzed the melting behavior of randomly selected human cDNA sequences obtained from genomic DNA by removing all introns. A clear correspondence is observed between exons and melting domains. This finding may provide new insights into the physical mechanisms underlying the evolution of genes. PMID- 15904338 TI - Optical conductivity of wet DNA. AB - Motivated by recent experiments, we study the optical conductivity of DNA in its natural environment containing water molecules and counterions. Our density functional theory calculations (using Siesta) for four base pair B-DNA with order 250 surrounding water molecules suggest a thermally activated doping of the DNA by water states which generically leads to an electronic contribution to low frequency absorption. The main contributions to the doping result from water near DNA ends, breaks, or nicks and are thus potentially associated with temporal or structural defects in the DNA. PMID- 15904339 TI - Divergence and Shannon information in genomes. AB - Shannon information (SI) and its special case, divergence, are defined for a DNA sequence in terms of probabilities of chemical words in the sequence and are computed for a set of complete genomes highly diverse in length and composition. We find the following: SI (but not divergence) is inversely proportional to sequence length for a random sequence but is length independent for genomes; the genomic SI is always greater and, for shorter words and longer sequences, hundreds to thousands times greater than the SI in a random sequence whose length and composition match those of the genome; genomic SIs appear to have word-length dependent universal values. The universality is inferred to be an evolution footprint of a universal mode for genome growth. PMID- 15904340 TI - Fluorescence fluctuation spectroscopy in subdiffraction focal volumes. AB - We establish fluorescence fluctuation spectroscopy (FFS) with nanoscale detection volumes generated by stimulated emission depletion. Our method applies fluorescence correlation spectroscopy and fluorescence intensity distribution analysis to extract molecular information about mobilities and fluorescence emission in solution. The combination of correlation analysis with that of photon intensity distributions reveals a fivefold squeezing of the detection volume over current diffraction-limited systems, which is in full agreement with the simultaneously demonstrated 25-fold reduction in (axial) focal transit time. Our method significantly extends the potential of far-field FFS, including for the noninvasive investigation of molecular reactions at higher concentrations. PMID- 15904341 TI - Miscibility gap in the microbial fitness landscape. AB - It is shown from molecular statistical considerations that a demixing instability exists in the moment space of a microbial protein expression profile. Although avoidance of demixing is generally requisite for biological function, a comparison with proteomic and genomic data suggests that many microbes lie close to the onset of this instability. Over evolutionary time scales, straying too close or into the immiscible domain may be associated with intracellular compartmentalization. PMID- 15904342 TI - Anomalous slowing down of the vibrational relaxation of liquid water upon nanoscale confinement. AB - We study the vibrational dynamics of nanodroplets of liquid water with femtosecond two-color midinfrared pump-probe spectroscopy. For the smallest nanodroplet, containing 10-15 water molecules, the lifetime T1 of the O-H stretch vibrations is equal to 0.85+/-0.1 ps, which is more than 3 times as long as in bulk liquid water. We find that the truncation of the hydrogen-bond network of water leads to a dramatic change of the relaxation mechanism. PMID- 15904343 TI - Voter model on heterogeneous graphs. AB - We study the voter model on heterogeneous graphs. We exploit the nonconservation of the magnetization to characterize how consensus is reached. For a network of N nodes with an arbitrary but uncorrelated degree distribution, the mean time to reach consensus T(N) scales as Nmu(2)1/mu(2), where mu(k) is the kth moment of the degree distribution. For a power-law degree distribution n(k) approximately k(-nu), T(N) thus scales as N for nu > 3, as N/ln(N for nu = 3, as N((2nu-4)/(nu 1)) for 2 < nu < 3, as (lnN)2 for nu = 2, and as omicron(1) for nu < 2. These results agree with simulation data for networks with both uncorrelated and correlated node degrees. PMID- 15904344 TI - Comment on "do intradot electron-electron interactions induce dephasing?". PMID- 15904346 TI - Formation dynamics of a fermion pair condensate. AB - The dynamics of pair condensate formation in a strongly interacting Fermi gas close to a Feshbach resonance was studied. We employed a phase-shift method in which the delayed response of the many-body system to a modulation of the interaction strength was recorded. The observable was the fraction of condensed molecules in the cloud after a rapid magnetic field ramp across the Feshbach resonance. The measured response time was slow compared to the rapid ramp, which provides final proof that the molecular condensates reflect the presence of fermion pair condensates before the ramp. PMID- 15904347 TI - Towards measuring variations of Casimir energy by a superconducting cavity. AB - We consider a Casimir cavity, one plate of which is a thin superconducting film. We show that when the cavity is cooled below the critical temperature for the onset of superconductivity, the sharp variation (in the far infrared) of the reflection coefficient of the film engenders a variation in the value of the Casimir energy. Even though the relative variation in the Casimir energy is very small, its magnitude can be comparable to the condensation energy of the superconducting film, and this gives rise to a number of testable effects, including a significant increase in the value of the critical magnetic field, required to destroy the superconductivity of the film. The theoretical ground is therefore prepared for the first experiment ever aimed at measuring variations of the Casimir energy itself. PMID- 15904348 TI - Systematically accelerated convergence of path integrals. AB - We present a new analytical method that systematically improves the convergence of path integrals of a generic N-fold discretized theory. Using it we calculate the effective actions S(p) for p< or =9, which lead to the same continuum amplitudes as the starting action, but that converge to that continuum limit as 1/N(p). We checked this derived speedup in convergence by performing Monte Carlo simulations on several different models. PMID- 15904349 TI - Unified and generalized approach to quantum error correction. AB - We present a unified approach to quantum error correction, called operator quantum error correction. Our scheme relies on a generalized notion of a noiseless subsystem that is investigated here. By combining the active error correction with this generalized noiseless subsystems method, we arrive at a unified approach which incorporates the known techniques--i.e., the standard error correction model, the method of decoherence-free subspaces, and the noiseless subsystem method--as special cases. Moreover, we demonstrate that the quantum error correction condition from the standard model is a necessary condition for all known methods of quantum error correction. PMID- 15904350 TI - Nature of the condensate in mass transport models. AB - We study the phenomenon of real space condensation in the steady state of a class of one-dimensional mass transport models. We derive the criterion for the occurrence of a condensation transition and analyze the precise nature of the shape and the size of the condensate in the condensed phase. We find two distinct condensate regimes: one where the condensate is Gaussian distributed and the particle number fluctuations scale normally as L(1/2) where L is the system size, and the second regime where the particle number fluctuations become anomalously large and the condensate peak is non-Gaussian. We interpret these results within the framework of sums of random variables. PMID- 15904351 TI - Experimental test of the fluctuation theorem for a driven two-level system with time-dependent rates. AB - A single defect center in diamond periodically excited by a laser is shown to provide a simple realization for a system obeying a fluctuation theorem for nonthermal noise. The distribution of these fluctuations is distinctly non Gaussian, which has also been verified by numerical calculation. For driving protocols symmetric under time reversal a more restricted form of the theorem holds, which is also known from entropy fluctuations caused by thermal noise. PMID- 15904352 TI - TeV mu neutrinos from young neutron stars. AB - Neutron stars are efficient accelerators for bringing charges up to relativistic energies. We show that if positive ions are accelerated to approximately 1 PeV near the surface of a young neutron star (t(age) less than or nearly 10(5) yr), protons interacting with the star's radiation field produce beamed mu neutrinos with energies of approximately 50 TeV that could produce the brightest neutrino sources at these energies yet proposed. These neutrinos would be coincident with the radio beam, so that, if the star is detected as a radio pulsar, the neutrino beam will sweep the Earth; the star would be a "neutrino pulsar." Looking for nu(mu) emission from young neutron stars will provide a valuable probe of the energetics of the neutron star magnetosphere. PMID- 15904353 TI - Massive graviton as a testable cold-dark-matter candidate. AB - We construct a consistent model of gravity where the tensor graviton mode is massive, while linearized equations for scalar and vector metric perturbations are not modified. The Friedmann equation acquires an extra dark-energy component leading to accelerated expansion. The mass of the graviton can be as large as approximately (10(15) cm)(-1), being constrained by the pulsar timing measurements. We argue that nonrelativistic gravitational waves can comprise the cold dark matter and may be detected by the future gravitational wave searches. PMID- 15904355 TI - Brane-induced-gravity shock waves. AB - We construct exact gravitational field solutions for a relativistic particle localized on a tensional brane in brane-induced gravity. They are a generalization of gravitational shock waves in 4D de Sitter space. We provide the metrics for both the normal branch and the self-inflating branch Dvali-Gabadadze Porrati brane worlds, and compare them to the 4D Einstein gravity solution and to the case when gravity resides only in the 5D bulk, without any brane-localized curvature terms. At short distances the wave profile looks the same as in four dimensions. The corrections appear only far from the source, where they differ from the long distance corrections in 4D de Sitter space. We also discover a new nonperturbative channel for energy emission into the bulk from the self-inflating [corrected] branch, when gravity is modified at the de Sitter radius. PMID- 15904356 TI - Direct proof of the tree-level scattering amplitude recursion relation in Yang mills theory. AB - Recently, by using the known structure of one-loop scattering amplitudes for gluons in Yang-Mills theory, a recursion relation for tree-level scattering amplitudes has been deduced. Here, we give a short and direct proof of this recursion relation based on properties of tree-level amplitudes only. PMID- 15904357 TI - From target to projectile and back again: self-duality of high-energy scattering evolution in QCD. AB - We prove that the complete kernel for the high-energy evolution in QCD must be self-dual. The relevant duality transformation is formulated in precise mathematical terms and is shown to transform the charge density into the functional derivative with respect to the single-gluon scattering matrix. This transformation interchanges the high and the low density regimes. We demonstrate that the original Jalilian-Marian-Iancu-McLerran-Weigert-Leonidov-Kovner kernel, valid at large density, is indeed dual to the low density limit of the complete kernel derived recently in hep-ph/0501198. PMID- 15904358 TI - Search for the lepton-number-violating decay Xi(-)-->pmu(-)mu(-). AB - A sensitive search for the lepton-number-violating decay Xi(-)-->pmu(-)mu(-) has been performed using a sample of approximately 10(9) Xi(-) hyperons produced in 800 GeV/c p-Cu collisions. We obtain B(Xi(-)-->pmu(-)mu(-))<4.0x10(-8) at 90% confidence, improving on the best previous limit by 4 orders of magnitude. PMID- 15904354 TI - Limits on gravitational-wave emission from selected pulsars using LIGO data. AB - We place direct upper limits on the amplitude of gravitational waves from 28 isolated radio pulsars by a coherent multidetector analysis of the data collected during the second science run of the LIGO interferometric detectors. These are the first direct upper limits for 26 of the 28 pulsars. We use coordinated radio observations for the first time to build radio-guided phase templates for the expected gravitational-wave signals. The unprecedented sensitivity of the detectors allows us to set strain upper limits as low as a few times 10(-24). These strain limits translate into limits on the equatorial ellipticities of the pulsars, which are smaller than 10(-5) for the four closest pulsars. PMID- 15904360 TI - Observation of B0-->pi0pi0. AB - We report the observation of the decay B0-->pi(0)pi(0), using a 253 fb(-1) data sample collected at the Upsilon(4S) resonance with the Belle detector at the KEKB e(+)e(-) collider. The measured branching fraction is B(B0- >pi(0)pi(0))=(2.3(+0.4+0.2)(-0.5-0.3))x10(-6), with a significance of 5.8 standard deviations including systematic uncertainties. We also make a measurement of the direct CP violating asymmetry in this mode. PMID- 15904363 TI - Equation of state of gluon plasma from a fundamental modular region. AB - Despite considerable practical success in dealing with the gluon plasma, finite temperature perturbation theory suffers at the fundamental level from infrared divergences discovered by Linde. However, if gauge or Gribov copies are properly eliminated from the physical state space, infrared modes are strongly suppressed. We describe the gluon plasma in zeroth order as a gas of free quasiparticles with a temperature-independent dispersion relation of Gribov type, E(k)=k(2)sqrt[+M4 / k(2)], that results from the reduction of the physical state space. The effective mass M2 / k controls infrared divergences and allows finite calculable corrections. The equation of state of this gas is calculated and compared with numerical lattice data. PMID- 15904364 TI - Proton electric pygmy dipole resonance. AB - The evolution of the low-lying E1 strength in proton-rich nuclei is analyzed in the framework of the self-consistent relativistic Hartree-Bogoliubov model and the relativistic quasiparticle random-phase approximation (RQRPA). Model calculations are performed for a series of N=20 isotones and Z=18 isotopes. For nuclei close to the proton drip line, the occurrence of pronounced dipole peaks is predicted in the low-energy region below 10 MeV excitation energy. From the analysis of the proton and neutron transition densities and the structure of the RQRPA amplitudes, it is shown that these states correspond to the proton pygmy dipole resonance. PMID- 15904362 TI - Observation of a near-threshold omegaJ/psi mass enhancement in exclusive B- >KomegaJ/psi decays. AB - We report the observation of a near-threshold enhancement in the omegaJ/psi invariant mass distribution for exclusive B-->KomegaJ/psi decays. The results are obtained from a 253 fb(-1) data sample that contains 275 x 10(6) BB pairs that were collected near the Upsilon(4S) resonance with the Belle detector at the KEKB asymmetric energy e(+)e(-) collider. The statistical significance of the omegaJ/psi mass enhancement is estimated to be greater than 8sigma. PMID- 15904365 TI - Time-dependent density functional theory of classical fluids. AB - We establish a rigorous time-dependent density functional theory of classical fluids for a wide class of microscopic dynamics. We obtain a stationary action principle for the density. We further introduce an exact practical scheme, to obtain hydrodynamical effects in density evolution, that is analogous to the Kohn Sham theory of quantum systems. Finally, we show how the current theory recovers existing phenomenological theories in an adiabatic limit. PMID- 15904366 TI - Observation of Feshbach resonances in an ultracold gas of 52Cr. AB - We have observed Feshbach resonances in collisions between ultracold 52Cr atoms. This is the first observation of collisional Feshbach resonances in an atomic species with more than one valence electron. The zero nuclear spin of 52Cr and thus the absence of a Fermi-contact interaction leads to regularly spaced resonance sequences. By comparing resonance positions with multichannel scattering calculations we determine the s-wave scattering length of the lowest (2S+1)Sigma(+)(g) potentials to be 112(14) a(0), 58(6) a(0), and -7(20) a(0) for S=6, 4, and 2, respectively, where a(0)=0.0529 nm. PMID- 15904367 TI - Generation of paired photons with controllable waveforms. AB - We describe experiments and theory showing the generation of counterpropagating paired photons with coherence times of about 50 ns and waveforms that are controllable at a rudimentary level. Using cw lasers, electromagnetically induced transparency and cold 87Rb atoms we generate paired photons into opposing single mode optical fibers at a rate of approximately 12 000 pairs per second. PMID- 15904368 TI - High-resolution ghost image and ghost diffraction experiments with thermal light. AB - High-resolution ghost image and ghost diffraction experiments are performed by using a single classical source of pseudothermal speckle light divided by a beam splitter. Passing from the image to the diffraction result solely relies on changing the optical setup in the reference arm, while leaving the object arm untouched. The product of spatial resolutions of the ghost image and ghost diffraction experiments is shown to overcome a limit which seemed to be achievable only with entangled photons. PMID- 15904369 TI - Weak localization of light in a disordered microcavity. AB - We report the observation of weak localization of light in a semiconductor microcavity. The intrinsic disorder in a microcavity leads to multiple scattering and hence to static speckle. We show that averaging over realizations of the disorder reveals a coherent backscattering cone that has a coherent enhancement factor > or =2, as required by reciprocity. The coherent backscattering cone is observed along a ring-shaped pattern due to confinement by the microcavity. PMID- 15904359 TI - Branching fractions and CP asymmetries in B0-->pi0pi0, B+-->pi+pi0, and B+- >K+pi0 decays and isospin analysis of the B-->pipi system. AB - Based on a sample of 227 x 10(6) BB pairs collected by the BABAR detector at the PEP-II asymmetric-energy B Factory at SLAC, we measure the branching fraction B(B0-->pi(0)pi(0))=(1.17+/-0.32+/-0.10)x10(-6), and the asymmetry Cpi(0)(pi(0))= 0.12+/-0.56+/-0.06. The B0-->pi(0)pi(0) signal has a significance of 5.0 sigma. We also measure B(B+-->pi(+)pi(0))=(5.8+/-0.6+/-0.4)x10(-6), B(B+- >K+pi(0))=(12.0+/-0.7+/-0.6)x10(-6), and the charge asymmetries Api(+)(pi(0))= 0.01+/-0.10+/-0.02 and AK+(pi(0))=0.06+/-0.06+/-0.01. Using isospin relations, we find an upper bound on the angle difference |alpha-alpha(eff)| of 35 degrees at the 90% C.L. PMID- 15904370 TI - Theory of mesoscopic magnetism in photonic crystals. AB - We provide a rigorous theoretical basis for the artificial magnetic activity of metamaterials near resonances. Our approach is a renormalization-based scheme that authorizes a completely general theory. The major result is an explicit expression of the effective permeability, in terms of resonant frequencies. The theoretical results are checked numerically, and we give applications of our theory to left-handed media and to the solution of the Pokrovski-Efros paradox. PMID- 15904361 TI - Measurement of the ratio of B+ and B0 meson lifetimes. AB - The ratio of the B+ and B0 meson lifetimes was measured using data collected in 2002-2004 by the D0 experiment in Run II of the Fermilab Tevatron Collider. These mesons were reconstructed in B-->mu(+)nuD(*-)X decays, which are dominated by B0 and B-->mu(+)nuD 0X decays, which are dominated by B+. The ratio of lifetimes is measured to be tau(+)/tau(0)=1.080+/-0.016(stat)+/-0.014(syst). PMID- 15904371 TI - Band gap formation and multiple scattering in photonic quasicrystals with a Penrose-type lattice. AB - This Letter presents a study of the local density of states (LDOS) in photonic quasicrystals. We show that the LDOS of a Penrose-type quasicrystal exhibits small additional band gaps. Among the band gaps, some exhibit a behavior similar to that typical of photonic crystals, while others do not. The development of certain band gaps requires large-size quasicrystals. It is explained by the long range interactions involved in their formation. Moreover, the frequencies where the band gaps occur are not necessarily explained using single scattering and should therefore involve multiple scattering. PMID- 15904372 TI - Velocity-selection problem for combined motion of melting and solidification fronts. AB - We discuss a free boundary problem for two moving solid-liquid interfaces that strongly interact via the diffusion field in the liquid layer between them. This problem arises in the context of liquid film migration (LFM) during the partial melting of solid alloys. In the LFM mechanism the system chooses a more efficient kinetic path which is controlled by diffusion in the liquid film, whereas the process with only one melting front would be controlled by the very slow diffusion in the mother solid phase. The relatively weak coherency strain energy is the effective driving force for LFM. As in the classical dendritic growth problems, also in this case an exact family of steady-state solutions with two parabolic fronts and an arbitrary velocity exists if capillary effects are neglected [D. E. Temkin, Acta Mater. 53, 2733 (2005)]. We develop a velocity selection theory for this problem, including anisotropic surface tension effects. PMID- 15904373 TI - Scaling and instabilities in bubble pinch-off. AB - We have used a 100 000 frame-per-second video to analyze the pinch-off of nitrogen gas bubbles in fluids with a wide range of viscosity. If the external fluid is highly viscous (eta(ext)>100 cP), the bubble neck radius is proportional to the time before break, tau, and decreases smoothly to zero. If the external fluid has low viscosity (eta(ext)<10 cP), the radius scales as tau(1/2) until an instability develops in the gas bubble, which causes the neck to rupture and tear apart. Finally, if the viscosity of the external fluid is in an intermediate range, an elongated thread is formed, which breaks apart into micron-sized bubbles. PMID- 15904374 TI - Two-dimensional solitons on the surface of magnetic fluids. AB - We report an observation of a stable solitonlike structure on the surface of a ferrofluid, generated by a local perturbation in the hysteretic regime of the Rosensweig instability. Unlike other pattern-forming systems with localized 2D structures, magnetic fluids are characterized by energy conservation; hence their mechanism of soliton stabilization is different from the previously discussed gain-loss balance mechanism. The radioscopic measurements of the soliton's surface profile suggest that locking on the wavelength defined by the nonmonotonic dispersion curve is instrumental in its stabilization. PMID- 15904375 TI - Faraday instability in a surface-frozen liquid. AB - Faraday surface instability measurements of the critical acceleration, a(c), and wave number, k(c), for standing surface waves on a tetracosanol (C24H50) melt exhibit abrupt changes at T(s)=54 degrees C, approximately 4 degrees C above the bulk freezing temperature. The measured variations of a(c) and k(c) vs temperature and driving frequency are accounted for quantitatively by a hydrodynamic model, revealing a change from a free-slip surface flow, generic for a free liquid surface (T>T(s)), to a surface-pinned, no-slip flow, characteristic of a flow near a wetted solid wall (T 0.05), but the scores of 'stomach' pain and 'stomach' pain before meals were higher in OD patients than in FD patients (2.65 +/- 1.11 vs 2.16 +/- 0.92, 2.26 +/- 1.26 vs 1.79 +/- 0.92, P < 0.05). In 45.2% of the OD patients and 47.7% of the FD patients, respectively, the severity of symptoms was not related to meals. In subgroups of ulcer-like, dysmotility-like and unspecified dyspepsia, the proportion of patients with symptoms not related to meals was 59.6%, 50.9% and 35.2%, respectively. 2.5% (6/239) of OD patients presented with progressive dysphagia, compared with 2.8% (15/543) of FD patients who presented with intermittent dysphagia. Approximately 8.8% (21/239) of OD patients reported dramatic weight loss accompanied with other severe symptoms, compared with 5.9% (32/543) of FD patients who had no other severe symptoms. A shift in symptom subtypes during the follow-up period was found in 13.8% of FD patients. The infection rate of Helicobacter pylori was higher in the OD group than in the FD group (53.1%vs 42.2%, P < 0.01), but no difference was found among the three subgroups of FD patients (P > 0.05). Halitosis was more often found in dyspeptic patients with H. pylori infection (44.9%vs 17.0% in OD, 47.3%vs 25.4% in FD, P < 0.01). CONCLUSIONS: When dyspepsia patients present with 'stomach' pain or 'stomach' pain before meals, a diagnosis of OD should be considered. Intermittent dysphagia, weight loss not accompanied with other severe symptoms, and halitosis (more often seen in patients with H. pylori infection) might be regarded as the relatively unique symptoms of dyspepsia in some FD patients. In FD, we found that the severity of dyspepsia symptoms was not related to meals in half of the patients, and symptom subtypes might shift over time, this adds difficulty to the management of FD. PMID- 15904424 TI - Synergistic effects of matrine and 5-fluorouracil on tumor growth of the implanted gastric cancer in nude mice. AB - OBJECTIVE: To investigate the inhibitive effects of matrine and 5-fluorouracil (5 FU) on the growth of human gastric adenocarcinoma cell line SGC-7901 when transplanted into nude mice and to investigate the bone marrow toxicity of these compounds. METHODS: 50 mg/kg and 100 mg/kg matrine with 50 mg/kg 5-FU, and 50 mg/kg matrine only, 50 mg/kg 5-FU only were intraperitoneally injected to observe their inhibitive effects by calculating the relative tumor volume (RTV) and tumor inhibition rates (IR%) as shown by the number of nucleated cells and bone marrow cell colony culture. RESULTS: The tumor inhibitive effect of the combined 100 mg/kg matrine and 50 mg/kg 5-FU group was stronger than that of the combined 50 mg/kg matrine and 50 mg/kg 5-FU groups (P < 0.05), and were also much stronger than that of the control group (50 mg/kg matrine only, 50 mg/kg 5-FU only, P < 0.01). As for the bone marrow inhibition effect, there was no significant statistical difference between the combination group and 5-FU alone group. In the cultured bone marrow cell colony, exuberant growth was seen in the combination group, but not in the control group. CONCLUSIONS: The inhibitive effect of combined matrine and 5-FU on the growth of transplanted human gastric cancer in nude mice is superior to that of matrine or 5-FU alone. Combined matrine and 5-FU can increase the inhibitive effect on proliferative hemopoietic bone marrow cells in nude mice and does not affect the resting bone marrow stem cells. PMID- 15904425 TI - Changes in gene expression profiles induced by parvovirus H-1 in human gastric cancer cells. AB - OBJECTIVE: The autonomous parvovirus H-1 exhibits preferential toxicity for transformed or tumor cells. The precise molecular mechanism of H-1 virus associated cytotoxicity is not fully understood. The present study aimed at gaining more information about parvovirus-induced cellular disturbances. METHODS: The H-1 virus-sensitive human gastric cancer cell line HGC27 was analyzed in the present study. cDNA microarrays were used to determine the global cellular gene expression changes which occur during the process of H-1 virus-induced death of HGC27 cells. A subset of differential expressed genes was further tested by RT PCR and Northern blot analyzes. RESULTS: A total of 920 genes belonging to various functional groups were found to be differentially expressed in H-1 virus- versus mock-infected cells in cDNA microarrays. Among them, 363 genes were upregulated, whilst 557 genes were downregulated. The differential expressions of some of these genes were further confirmed by RT-PCR and Northern blot analysis. CONCLUSION: Some of genes known to be involved in cell signal transduction, apoptosis, DNA replication, DNA repair, DNA binding and transcription were differentially expressed after parvovirus H-1 infection, they might play a role in H-1 virus-induced gastric cancer cell death. These genes represent interesting candidates to be tested at the functional level for their contribution to the disturbances triggered by H-1 virus in tumor cells. PMID- 15904426 TI - Modulation of multidrug resistance by andrographolid in a HCT-8/5-FU multidrug resistant colorectal cancer cell line. AB - OBJECTIVE: To develop an HCT-8/5-FU multidrug-resistant colorectal cancer cell line and to elucidate the effect of Andrographolid (AG), an extract from Andrographis paniculate, a medicinal herb on the HCT-8/5-FU multidrug-resistant colorectal cancer cell line. METHODS: An HCT-8 colorectal cancer cell line was used and a high concentration of 5-Fluorouracid (5-FU) was introduced at the beginning to induce drug resistance, then the concentration of 5-FU was increased in gradients. Approximately 7 months later, the cells grew stably in 2.0 microg/mL of 5-FU, and the cell line was named HCT-8/5-FU multidrug-resistant colorectal cancer cell line. The resistant index of HCT-8/5-FU cells to 5-FU, adriamycin (ADM), cisplatin (DDP) was checked by MTT test, and a growth curve was drawn. The morphological changes were observed by both light and electron microscope. The function of P-170 was detected by rhodamine staining. After the application of AG and co-administration of 5-FU, ADM and DDP, the growth curves and inhibition rate as well as apoptosis rate of HCT-8/5-FU at different concentrations of AG were evaluated by MTT and flow cytometry. Rhodamine staining was used to investigate the possible mechanism involved by AG. RESULTS: The resistance index of HCT-8/5-FU to 5-FU was 16.6, and a cross-resistance to ADM and DDP was noticed. Compared with parental cells, HCT-8/5-FU cell's growth rate did not change significantly but the cell's morphology was remarkably changed as compared with parental cells. Overexpression of P-170 by HCT-8/5-FU cell was indicated through rhodamine staining. AG at a low concentration showed weak inhibitory effect on HCT-8/5-FU. However, a remarkable inhibitory and apoptosis rate was shown when AG was co-administered with 5-FU, ADM and DDP, respectively. Interestingly AG alone could not induce apoptosis and change the cell cycles. AG might affect the expression of P-170, which was indicated by rhodamine staining. CONCLUSIONS: The HCT-8/5-FU multidrug-resistant colorectal cancer cell line has been successfully developed and because it has cross-resistance to 5-FU, ADM and DDP, it might serve as an ideal multidrug resistance (MDR) model for colorectal cancer research. The mechanism of HCT-8/5-FU resistance to chemotherapeutic agents might be related to the overexpression of P-170. Low concentrations of AG alone have no significant inhibition on HCT-8/5-FU and fail to induce apoptosis and to change cell cycles. AG might act as a chemosensitizer when co-administered with 5-FU, ADM and DDP, and the mechanism of reversal modulation of multidrug resistance by AG in the HCT-8/5-FU resistant cell line might be related to its downregulation of overexpression of P-170. PMID- 15904427 TI - Diagnosis and management of regional portal hypertension. AB - OBJECTIVE: Regional portal hypertension (RPH) results from splenic vein thrombosis/occlusion, with emergence of gastric varices and severe upper gastrointestinal bleeding. Its diagnosis and management are summarized below. METHODS: We reviewed our experience in 16 consecutive patients with RPH at Chinese PLA General Hospital between 2001 and 2004. The clinical manifestations, diagnostic methods and therapeutic modalities were analyzed retrospectively. RESULTS: Among the 16 patients with RPH, 12 had co-existing pancreatic diseases and 4 were of obscure origin. The main clinical findings were splenomegaly in 16 (100%), epigastric pain (mainly left upper abdomen) in 10 (63%), gastrointestinal bleeding in 7 (44%) and abdominal mass in 3 (19%). All had normal liver function tests. The main diagnostic methods used were ultrasound (US), computerized tomography (CT) and endoscopy. Splenic vein thrombosis could be identified by color Doppler ultrasound (7/7), enhanced CT could show its enlarged and tortuous short gastric vein, gastroepiploic vein, and collateral vessels around the splenic hilum (16/16). Isolated gastric varices (4/5) were mainly shown by esophagogastroscopy. Splenectomy was effective in controlling the variceal bleeding, but portal vein thrombosis occurred in 1 case. Endoscopic sclerotherapy had been carried out repeatedly in 2 cases, but was less effective (1/2, 50%). CONCLUSIONS: Precise diagnosis of RPH can be achieved by clinical manifestations, US and CT. Therapeutic options should be individualized basing on the underlying diseases, however, splenectomy is more preferable for patients with bleeding varices, and endoscopic sclerotherapy can be tried. PMID- 15904428 TI - Effect of proapoptosis protein on hepatocarcinogenesis. AB - OBJECTIVE: The goal of the present study was to determine how proapoptosis proteins regulate the progression of liver proliferative foci and tumorigenesis initiated by a chemical carcinogen, diethylnitrosamine (DEN). METHODS: Bid deficient mice (15-day-old) were injected with 15 microg/g of DEN, then killed at 3 and 10 days, and 4 and 8 months after injection for analysis of hepatocellular proliferation, apoptosis and tumorigenesis. RESULT: The rate of apoptosis in the hepatocytes of the wild-type mice was significantly higher than in the Bid deficient mice at 10 days after DEN exposure (P < 0.0001); the results of BrdU labeling agreed with the measurement of apoptosis in these animals, showing an obvious increase in the wild-type mice compared with the Bid-deficient mice (P < 0.0001). Four months after DEN exposure, the number and size of lesion foci or nodules in the wild-type mice were both greater than in the Bid-deficient mice (P < 0.05 and P < 0.001, respectively), but there was no significant difference between the two groups of mice at 8 months. CONCLUSION: These results suggest that a lack of apoptosis in liver tissue in the early stage after DEN exposure decreased some of the tumorigenesis potential of DEN. PMID- 15904429 TI - In view of an optimal gut antifungal therapeutic strategy: an in vitro susceptibility and toxicity study testing a novel phyto-compound. AB - OBJECTIVE: In view of the raising concern for gut fungal infection, the aim of the present research was to carry out a systematic in vitro study testing the antifungal activity and possible toxicity of a polygodyal-anethole compound (Kolorex) in several strains of Candida albicans and in other fungal pathogens. METHODS: The in vitro susceptibility tests were carried out on 4 strains of C. albicans (C. krusei, C. lipolytica, C. tropicalis, C. utilis), Aspergillus flavus and A. fumigatus. Cultures were also analyzed by varying medium, pH and inoculum size, and a time-course killing test was carried out. RESULTS: In the present study the polygodyal-anethole compound showed remarkable in vitro activity against the most common fungi, which was significantly better than polygodyal alone. Moreover, such mixture compound was shown to exert its activity against a wide spectrum of fungi, including C. lipolytica and C. tropicalis, which required significantly higher MIC of polygodyal to be unfeasible in clinical application. The activity of the polygodyal-anethole compound was significantly better than polygodyal alone with high inoculum size and low pH. Moreover, it proved to exert a significantly faster biological activity against low inoculum. CONCLUSIONS: This study suggests that the mixture compound Kolorex has a very good profile of antifungal activity in terms of effectiveness and spectrum of action while being devoid of any significant toxicity. PMID- 15904430 TI - Assisted reproductive technologies and birth defects. AB - In vitro fertilization (IVF) and other assisted reproductive technologies (ART) are effective treatments for infertility and are widely provided at infertility clinics. Although IVF and related ART procedures are generally considered safe, some studies have suggested an excess occurrence of major malformations, low birth-weight and other perinatal complications in babies conceived by ART. Further, it was recently reported that IVF and intracytoplasmic sperm injection (ICSI) are associated with imprinting disorders in the offspring such as Beckwith Wiedemann syndrome and Angelman syndrome. Here we review the human and animal studies investigating the potential risks of ART, and discuss the need for further investigation. PMID- 15904431 TI - Methodological issues in the epidemiological study of the teratogenicity of drugs. AB - The review presented here discusses and exemplifies problems in epidemiological studies of drug teratogenesis according to methodology: case-control studies, cohort studies, or total population studies. Sources of errors and the possibility of confounding are underlined. The review stresses the caution with which conclusions have to be drawn when exposure data are retrospective or other possible bias exists. It also stresses the problem with the multiple testing situation that is usually present in the studies. It is therefore difficult to draw any firm conclusion from single studies and still more difficult to draw conclusions on causality. As randomized studies are in most cases out of the question, one has to rely on the type of studies which can be made, but the interpretation of the results should be cautious. The ideal study, next to a randomized one, is a large prospective study with detailed exposure information and detailed and unbiased outcome data. Even so, such a study can mainly be used for identifying possible associations which have to be verified or rejected in new studies. Nearly every finding of a risk increase, if not extremely strong, should only be regarded as a tentative signal to be tested in independent studies. PMID- 15904432 TI - Morphological differences in cardiovascular anomalies induced by bis-diamine between Sprague-Dawley and Wistar rats. AB - It is known that animals show different responses to the same teratogen between different strains. We examined cardiac malformations in Sprague-Dawley (SD) and Wistar rats induced by bis-diamine, which produced conotruncal anomalies and aortic arch malformations in embryos when administered to the dams, to elucidate the morphological differences and pathogenesis in the two strains. Two hundred milligrams of bis-diamine dissolved in 1% gum-tragacanth was administered to pregnant rats on embryonic day (ED) 9.5, 10.5 and 11.5 in each strain. The embryos were removed on ED 20.5. External appearances, cardiovascular morphology and associated anomalies were examined under a dissecting microscope. An immunohistological study with an anti-N-CAM antibody, an excellent marker for neural crest cells, was performed on ED 12.5 embryos. Isolated aortic arch anomalies were common features of malformations induced by bis-diamine in SD rats and intracardiac defects were found in a small number of the embryos. Wistar rats showed more serious cardiovascular anomalies, such as persistent truncus arteriosus and tetralogy of Fallot, especially when dams were treated on ED 10.5 and isolated arch anomalies were significantly less prevalent than in SD rats. Immunohistology demonstrated that there were fewer N-CAM positive cells in the conotruncal region in Wistar rats than in SD rats. Bis-diamine induced more critical cardiovascular malformations in Wistar rats because neural crest cells, which play an important role in conotruncal septation, were more extensively damaged. Different susceptibility to bis-diamine and/or different time of neural crest cell emigration from the hindbrain might explain those morphological differences. PMID- 15904433 TI - Clinical variability in a Japanese hereditary lymphedema type I family with an FLT4 mutation. AB - Hereditary lymphedema type I (Milroy disease) is a rare autosomal dominant disease resulting from mutations of FLT4 encoding the vascular endothelial growth factor receptor-3. Patients develop edema of the legs and feet, resulting in chronic swelling of the lower extremities from the neonatal period. Here we report a Japanese family with 10 affected members of five generations of hereditary lymphedema type I. We identified a previously reported missense mutation of G857R in one allele of FLT4 from three affected individuals of three generations, the mother of whom presented only hemi-lymphedema of the left foot. Thus, the clinical features of hereditary lymphedema type I caused by a FLT4 mutation are heterogeneous and it would be appropriate to consider FLT4 mutations even in a patient with hemi-lymphedema of the foot. PMID- 15904434 TI - Refining chromosomal region critical for Down syndrome-related heart defects with a case of cryptic 21q22.2 duplication. AB - We report here a patient with features of Down syndrome and tetralogy of Fallot who had a 21q22 duplication. The extent of the duplication was defined using fluorescent hybridization probes that map to the critical region on chromosome 21. Included within the interval was the cell adhesion molecule DSCAM but not the collagen COL6A1. The present case provides further support to the concept that there exists Down syndrome-associated congenital heart disease gene(s) on chromosome 21q22 and that over-expression of DSCAM may contribute to the cardiac defects of Down syndrome. PMID- 15904435 TI - Rubinstein-Taybi syndrome (RTS) with postaxial polydactyly of the foot: 4-year follow-up until improvement of dysbasia. AB - Rubinstein-Taybi syndrome (RTS), also known as 'broad thumbs syndrome' or 'broad thumb-hallux syndrome', is a malformation syndrome characterized by the triad of broad thumbs or first toes, a peculiar facial expression called 'comical face' and mental retardation. Although various malformations are combined with the triad, polydactyly is rare. We treated a male patient with RTS complicated by postaxial polydactyly of the foot. His clinical course was different from typical patients with polydactyly, especially in the aspect of walking development. Osteoplasty-combined surgery, which was ideal for anatomical reconstruction, was performed on the patient at 2 years and 11 months of age. A 4-year follow-up period was required until there was an improvement of dysbasia. PMID- 15904436 TI - First case of L1CAM gene mutation identified in MASA syndrome in Asia. AB - We report here the first case of an L1CAM gene mutation identified in mental retardation, adducted thumbs, shuffling gait, and aphasia (MASA) syndrome in Japan. The patient was a 10-year-old boy with mild mental retardation, bilateral adducted thumbs and corpus callosum hypoplasia. His family had no history of MASA syndrome. The L1CAM gene contained a nonsense mutation (R1166X) in exon 26 in the cytoplasmic domain. No mutation was found in the extracellular and transmembrane domains of L1CAM. The abnormal development of axon tracts resulting in the corpus callosum hypoplasia and adducted thumbs appears to be caused by malfunction of the cytoplasmic domain of L1CAM. PMID- 15904437 TI - Acardiac acephalic twin gestation with transposition of great arteries in pump twin. AB - We report here a case of twin reversed arterial perfusion sequence with transposition of great arteries in the pump twin. Color Doppler and fetal echocardiographic examination revealed an acardiac, anencephalic twin without upper limbs and a pump twin with transposition of great arteries. To our knowledge despite multiple anomalies reported in the acardiac twin, major anomalies, especially of a cardiac type are extremely rare and not reported in the pump twin, which maintains circulation. Management is directed at saving the pump fetus; therefore, we believe that during sonographic exam, emphasis should be placed on associated organ anomalies or complications that may hamper viability of the pump. PMID- 15904439 TI - A rare 19th century French obstetrical atlas. PMID- 15904440 TI - Vaginal birth after classical Caesarean section. PMID- 15904441 TI - Paravaginal defects: a comparison of clinical examination and 2D/3D ultrasound imaging. AB - BACKGROUND: Paravaginal defects are often assumed to be the underlying anatomical abnormality in anterior compartment descent. Neither clinical examination nor ultrasound assessment are generally accepted diagnostic modalities. AIMS: To compare clinical examination and translabial 3D ultrasound in the detection of such defects. METHODS: Fifty-nine women without previous prolapse or incontinence surgery were seen prospectively. Clinical and ultrasound assessments were carried out in blinded fashion. 3D translabial ultrasound was undertaken after voiding and supine. Volumes were acquired at rest, on Valsalva and on levator contraction. Loss of paravaginal support ('tenting') in the axial plane was taken to signify paravaginal defects. RESULTS: Paravaginal defects were reported clinically in 14 cases on the left (24%), 19 times on the right (32%). Two 3D ultrasound examinations did not yield satisfactory volumes, leaving 57 for analysis. Neither midsagittal nor coronal views yielded data that correlated with clinical assessments. In the axial plane there was absence of tenting at rest in 32/57 (57%) patients, but this did not correlate with clinical findings. Loss of tenting on Valsalva was observed less often (21/57, 37%) and was weakly associated with clinically observed lateral defects (P = 0.036). CONCLUSIONS: Pelvic floor ultrasound in midsagittal, axial or coronal planes does not correlate well with clinical assessment for paravaginal defects. This could be due to poor clinical assessment technique or limitations of the ultrasound method. On the other hand, paravaginal defects may be uncommon or clinically irrelevant. On present knowledge, the paravaginal defect has to be regarded as an unproven concept. PMID- 15904442 TI - Transperineal versus transvaginal ultrasonographic evaluation of the cervix at each trimester in normal pregnant women. AB - AIMS: To compare transvaginal and transperineal ultrasonography in the assessment of cervical length and cervical changes in normal gravid patients at each trimester. METHODS: Transperineal and transvaginal ultrasonographic cervical length was measured on 104 asymptomatic pregnant women between 10 and 14, 20-24, and 30-34 weeks' gestation and the presence of a funnel was also noted. The study used the McNemar chi2 test to assess the difference between two methods in their ability to obtain a measurement, and the Pearson correlation coefficient to determine the relationship between the paired transperineal and transvaginal cervical lengths. RESULTS: Cervical length measurements were obtained by transvaginal ultrasonography in all 104 patients and by transperineal ultrasonography in 101 patients (97.1%) (P = 0.1). By gestational age, the greatest length discrepancy (2.8 mm) between the two ultrasonographic methods was found at 10-14 weeks (P < 0.001). At 20-24 and 30-34 weeks' gestation, the mean length differences were less than 1 mm (P < 0.01 and P = 0.337, respectively). Cervical funnelling was observed in 16 patients by both methods, whereas in two patients from the 20-24 week gestational age group, funelling was observed by transvaginal ultrasonography and not by transperineal ultrasonography. CONCLUSIONS: Cervical length measurements by transperineal ultrasonography show good correlation with transvaginal ultrasonographic measurements and it is a satisfactory alternative to a transvaginal evaluation of the cervix throughout pregnancy. PMID- 15904443 TI - Bowel, bladder and sexual function in women undergoing laparoscopic posterior compartment repair in the presence of apical or anterior compartment dysfunction. AB - OBJECTIVE: The aim of the study was to analyse the functional outcome of women undergoing a laparoscopic posterior compartment repair in the presence of anterior or apical compartment dysfunction. DESIGN: Prospective cohort study. METHODS: Forty women, median age 65 years (41-78), with symptoms of genital prolapse 31 (78%), urinary dysfunction 32 (80%) and bowel dysfunction 40 (100%), underwent laparoscopic posterior compartment repair in conjunction with an anterior compartment repair. Pre-operative and postoperative bowel and bladder function was prospectively assessed with a Wexner continence score, Vienna constipation score and a urinary dysfunction score. Twenty-eight (70%) and 24 patients (60%) had pre-operative urodynamics and anorectal manometry. Post operatively all women were also assessed with a Watt's sexual dysfunction score and a linear analogue patient satisfaction score. Twelve women (30%) had postoperative anal manometry. RESULTS: At 20 months median follow-up, 30 (97%), 20 (62%) and 12 (31%) women reported improvement in their prolapse, urinary and bowel symptoms, respectively. Post-operatively, one woman reported denovo faecal incontinence, four worsening obstructive defecation and three denovo urinary dysfunction. Nine women (35%) reported denovo dyspareunia. The mean time to clinical deterioration following surgery was 11 months. Bowel function improvement was the only factor to significantly correlate with postoperative patient satisfaction. CONCLUSION: The functional outcome of laparoscopic posterior compartment repair in the presence of anterior compartment dysfunction is disappointing. Preoperative counselling is important to ensure that patients have reasonable and realistic expectations from repair surgery, and an understanding that anatomical improvement might not be followed by long-term functional improvement. PMID- 15904444 TI - Management of preterm prelabour rupture of membranes: an audit. How do the results compare with clinical practice guidelines? AB - BACKGROUND: Preterm prelabour rupture of membranes is a common clinical event. It is associated with infection in approximately 50% of cases. Clinical practice guidelines have been developed at the Royal Women's Hospital, Melbourne, Australia for investigation and management of this condition. AIM: To perform an audit of management of women presenting with this diagnosis and assess how inpatient management compares with the Hospital's current clinical practice guideline and how the clinical practice guideline compares with the evidence in the literature. METHODS: Retrospective audit over a 3-month period collecting data on maternal age, gestation, microbiological results, other investigations, pharmacological treatment and outcome. RESULTS: All the 56 women admitted for this reason received at least one dose of antibiotic, most commonly erythromycin. More than two thirds of patients had the antibiotic changed at least once during their admission. Ten patients were prescribed intravenous antibiotics without a clear indication. Sixty-four percent received steroids for lung maturation of the neonate and 30% received tocolysis with nifedipine. Almost two thirds of patients delivered within 7 days and there were four neonatal deaths. CONCLUSION: In general management of women with premature rupture of membranes is in keeping with the current clinical practice guideline at the Royal Women's Hospital although antibiotic prescribing and management of Group B streptococcus colonisation could be improved. In addition, routine measurement of C reactive protein should cease. The current clinical practice guideline should be modified to reflect the current evidence in the literature. PMID- 15904445 TI - Antenatal antecedents of moderate or severe neonatal encephalopathy in term infants--a regional review. AB - BACKGROUND: Regional audits of term infants with neonatal encephalopathy (NE) provide an opportunity to examine issues related to causation and quality of care. AIM: To document antenatal and intrapartum antecedents in a contemporary cohort of term infants with moderate or severe neonatal encephalopathy. METHODS: Term infants admitted with moderate-severe neonatal encephalopathy over 4 years were identified. The clinical records were reviewed for information about the pregnancy and birth including interpretation of monitoring and subsequent management of the labour and delivery. RESULTS: Fifty-two maternal records were reviewed. No mothers were diabetic or had gestations > 42 weeks, but 17% of the babies were small for gestational age (SGA). The cohort had evidence of antenatal hypoxia in 15%, a sentinel event in 25% and suboptimal fetal monitoring practice in at least 42% of cases. CONCLUSIONS: Peripartum events were the major contributors to neurological damage in infants with neonatal encephalopathy. Suboptimal fetal monitoring practice and sentinel events remain the most common contributors. Ongoing education and training to address these issues should be available to all involved with intrapartum care in New Zealand. PMID- 15904446 TI - Presenting symptoms of epithelial ovarian cancer. AB - OBJECTIVES: Epithelial ovarian cancer is a common disease with a high mortality, the latter being frequently attributed to late diagnosis due to failure to recognise symptoms of early disease. This study was designed to determine any differences in symptomatology between patients with early and advanced stage disease. DESIGN: A retrospective cohort study of 100 patients with stage 1 disease and 100 with stage 3 disease. Ten consecutive patients with stage 1 disease and 10 with stage 3 disease were identified from the database of patients treated each year from 1990 to 1999. SETTING: A tertiary referral centre for gynecological malignancy. OUTCOME MEASURES: The nature and duration of symptoms, age at presentation, and the size, histologic type and grade of tumour were determined by chart review, and the findings compared using logistic regression analysis. RESULTS: Ninety percent of women with early, and 100% with advanced disease reported at least one symptom. With early disease, abdominal pain was reported by 51% and abdominal swelling by 32%, and with advanced disease abdominal swelling by 62% and abdominal pain by 44%. Seventy percent of the early stage and 69% of the advanced stage cohorts reported symptoms of less than 3 months duration. Tumours less than 5 cm diameter were three times more likely to have advanced disease (P = 0.02). Grade 1 tumours were 40 times more likely to be early stage than grade 3 tumours. Serous tumours occurred in 25% of patients with early disease, and 45% with advanced disease (P = 0.01). CONCLUSION: Patients with early stage ovarian cancer are likely to be younger and to have larger, better differentiated tumours that are more often non-serous histologically. Advanced disease is not invariably due to delayed diagnosis. PMID- 15904447 TI - Comparison of a self-administered tampon ThinPrep test with conventional pap smears for cervical cytology. AB - AIM: To assess a self-administered tampon specimen as an alternative method of detecting cytological abnormalities and its acceptability in comparison with a conventional Papanicolou (pap) smear. DESIGN: Comparative observational study. SETTING/POPULATION: Two hundred and seventeen women were recruited from the colposcopy clinic of an outer urban public teaching hospital and from sexual health clinics at suburban and major metropolitan hospital clinics. METHODS: Participants inserted and immediately withdrew a tampon, then placed it into a vial of ThinPrep PreservCyt fluid. This was analysed by a local private pathology laboratory. Results were compared to a pap smear performed the same day or within the previous 6 months. All women with an abnormal result (tampon or pap smear) underwent a colposcopy, with or without biopsy as necessary. Participants completed a questionnaire after performing the tampon test. OUTCOME MEASURES: Probabilities of tampon test detecting (i) a high grade abnormality (pHG), (ii) any cervical intraepithelial neoplasia (CIN) changes (pCINany), and (iii) any abnormalities (pabn) compared to the conventional pap smear and, if abnormal, compared to the biopsy taken at colposcopy. Acceptability of the tampon test and conventional pap smear were also measured. RESULTS: Probabilities of the tampon test compared to pap smear: pabn sensitivity 33%, specificity 89%, PPV 59%, NPV 73%; pCINany sensitivity 23%, specificity 97%, PPV 71%, NPV 79%; pHG sensitivity 19%, specificity 98%, PPV 63%, NPV 89%. Acceptability for tampon test was 91.21% and for pap smear, 45.85%. CONCLUSIONS: Although the self-administered tampon ThinPrep method is a poor detector of cervical abnormalities compared to pap smear, it is highly acceptable to women. It has a relatively good negative predictive value (NPV). Our study suggests that if a more acceptable, sensitive method of cervical screening was found, which removed some of the existing barriers to conventional pap testing, screening rates for cervical cancer may improve. PMID- 15904448 TI - Substance use during pregnancy: risk factors and obstetric and perinatal outcomes in South Australia. AB - OBJECTIVE: To determine the prevalence of self-reported substance use during pregnancy in South Australia, the characteristics of substance users, their obstetric outcomes and the perinatal outcomes of their babies. METHODS: Multivariable logistic regression with STATA statistical software was undertaken using the South Australian perinatal data collection 1998-2002. An audit was conducted on every fifth case coded as substance use to identify the actual substances used. RESULTS: Substance use was reported by women in 707 of 89 080 confinements (0.8%). Marijuana (38.9%), methadone (29.9%), amphetamines (14.6%) and heroin (12.5%) were most commonly reported, with polydrug use among 18.8% of the women audited. Substance users were more likely than non-users to be smokers, to have a psychiatric condition, to be single, indigenous, of lower socio economic status and living in the metropolitan area. The outcome models had poor predictive powers. Substance use was associated with increased risks for placental abruption (OR 2.53) and antepartum haemorrhage from other causes (OR 1.41). The exposed babies had increased risks for preterm birth (OR 2.63), small for gestational age (OR 1.79), congenital abnormalities (1.52), nursery stays longer than 7 days (OR 4.07), stillbirth (OR 2.54) and neonatal death (OR 2.92). CONCLUSIONS: Substance use in pregnancy is associated with increased risks for antepartum haemorrhage and poor perinatal outcomes. However, only a small amount of the variance in outcomes can be explained by the substance use alone. Recent initiatives to improve identification and support of women exposed to adverse health, psychosocial and lifestyle factors will need evaluation. PMID- 15904449 TI - Physiotherapy for female stress urinary incontinence: a multicentre observational study. AB - BACKGROUND: No previous data are available on the effectiveness of physiotherapy management of urinary stress incontinence with relevance to the Australian health system. AIMS: To evaluate Australian ambulatory physiotherapy management of stress urinary incontinence. METHODS: Observational multicentre clinical study of physiotherapy management of female stress urinary incontinence between February 1999 and October 2000, with 1-year follow-up. Outcome measures were a stress test and a 7-day diary of incontinent episodes (pretreatment and at every visit) and a condition-specific quality of life (QoL) questionnaire (pre- and post-treatment). Subjects were followed-up 1 year after treatment by questionnaire with a 7-day diary, QoL questionnaire, and assessment of subjective outcome, subjective cure, satisfaction and need for surgery. RESULTS: Of the 274 consenting subjects, 208 completed an episode of physiotherapy care consisting of a median (IQ range) of five (four to six) visits. At the end of the episode, 84% were cured and 9% improved on stress testing, whilst 53% were cured and 25% improved according to the 7-day diary. Mean volume of urine loss on stress testing reduced from 2.4 (2.5) mL to 0.1(0.4) mL after treatment. There was a significant improvement in all QoL domains. Median (interquartile range) incontinent episodes per week were reduced from five (three to 11) to zero (zero to two) (P < 0.05) after treatment and to one (zero to four) at 1 year (P < 0.05). At 1 year, approximately 80% of respondents had positive outcomes on all outcome measures. CONCLUSIONS: Physiotherapy management in Australian clinical settings is an effective treatment option for women with stress urinary incontinence. PMID- 15904450 TI - Primary postpartum haemorrhage in an Australian tertiary hospital: a case-control study. AB - The present study aimed to determine the incidence of primary postpartum haemorrhage (PPH) after vaginal birth at an Australian tertiary hospital, and to investigate risk factors for primary PPH at this hospital. A case-control study of women delivering vaginally at a tertiary hospital from February to June 2003 was performed. Demographic, antenatal, intrapartum, treatment and outcome data were abstracted from patient records. The study population comprised 125 cases and 125 controls, with a primary PPH rate of 12.1 per 100 vaginal births. Risk factors on multivariate analysis were past history of PPH, second stage labour > 60 min, forceps delivery, and incomplete placenta/ragged membranes. PMID- 15904451 TI - Use of urethral plugs for urinary incontinence following fistula repair. AB - A common complication following anatomical closure of obstetric genito-urinary fistula is urinary incontinence. Management is often suboptimal with lack of urodynamic equipment in most fistula centres in developing countries. Surgical interventions have been described with varying success. The aim of this paper is to describe the use of urethral plugs as an alternative management for women with postfistula incontinence, in a developing country. A pilot study was undertaken to assess the effectiveness of the urethral plugs in these women. The use of urethral plugs appear to be an effective short-term management of women with postfistula incontinence, with minimal complications. Longer follow-up and in larger numbers are required. PMID- 15904452 TI - Outcomes of pregnancy complicated by thyroid disease. AB - AIMS: To perform a case note review of pregnancies complicated by thyroid dysfunction to determine management and therapeutic intervention in relation to pregnancy outcome. METHODS: A retrospective case note analysis of 81 ongoing pregnancies in 70 pregnant women with a history of thyroid dysfunction over a period of 5 years at the Glasgow Royal Maternity Hospital (GRMH), Glasgow, Scotland, United Kingdom. The results of thyroid function tests and whether a change in treatment was instituted were recorded. Thyroid function was assessed by standard laboratory reference ranges for free thyroxine (FT4) and thyroid stimulating hormone (TSH) in all trimesters. Other parameters were also noted. RESULTS: Medication levels needed to be increased in the hypothyroid group (45%), and decreased (38%) in the hyperthyroid group. CONCLUSION: Pregnancy outcome was good in majority of cases given appropriate replacement therapy for stated reference values. PMID- 15904453 TI - Pregnancy and the Rastelli operation. AB - BACKGROUND: The aim of surgical correction of congenital heart disease is for the patient to live a relatively normal life, including potential pregnancy and childbirth. The Rastelli repair operation using a valved conduit for certain complex congenital heart lesions has resulted in survival into adult life. AIMS: We reviewed reproductive outcome of 22 women over 18 years of age who were survivors of the Rastelli operation performed between 1977 and 1999. RESULTS: Six women had a total of 12 pregnancies with six live births. Three women, with complex transposition of the great arteries, developed more severe subaortic obstruction during pregnancy. All required further heart surgery. Delivery was by Caesarean section in three patients and vaginally in the other three. All infants were healthy and free from defects. Two women had tubal ligations because of their cardiac problems. CONCLUSIONS: Women who have had Rastelli repair operation for complex congenital heart disease can have successful pregnancies. Subaortic stenosis can become a problem and should ideally be eliminated before pregnancy. Because of multiple operations, complications and genetic concerns, some women choose not to reproduce. Careful cardiac evaluation before, during and after pregnancy is essential. PMID- 15904454 TI - Unexpected complication of uterine myoma morcellation. PMID- 15904455 TI - Sheehan's syndrome presenting post-partum with diabetes insipidus. PMID- 15904456 TI - A new surgical technique for dealing with uterine inversion. PMID- 15904457 TI - Pityriasis rosea in pregnancy--specific diagnostic implications and management considerations. PMID- 15904458 TI - Isolated vulval varicosity in the non-pregnant state: a case report with review of the treatment options. PMID- 15904459 TI - Life-threatening uterine haemorrhage six weeks after Caesarean section due to uterine scar endometriosis: case report and review of literature. PMID- 15904460 TI - Re: Severe hyponatraemia as a result of primary polydipsia in labour. PMID- 15904461 TI - Re: Severe hyponatraemia as a result of primary polydipsia in labour. PMID- 15904462 TI - Why are we still doing radical hysterectomies for cervical cancer? PMID- 15904464 TI - Tumor immunity against adult T-cell leukemia. AB - Human T-cell leukemia virus type-I (HTLV-I) causes adult T-cell leukemia (ATL) in a small population of infected individuals after a long incubation period. Although the process of clonal evolution of ATL cells may involve multiple steps, ATL cells from half of the ATL cases still retain the ability to express HTLV-I Tax, a key molecule of HTLV-I leukemogenesis. A recent finding of reactivation of Tax-specific cytotoxic T lymphocytes (CTL) in ATL patients after hematopoietic stem cell transplantation suggests the presence of Tax expression in vivo and potential contribution of the CTL to antitumor immunity. This is consistent with the results of a series of animal experiments indicating that Tax-specific CTL limit the growth of HTLV-I-infected cells in vivo, although the animal model mimics only an early phase of HTLV-I infection and leukemogenesis. Establishment of an insufficient HTLV-I-specific T-cell response and an increased viral load in orally HTLV-I-infected rats suggests that host HTLV-I-specific T-cell response at a primary HTLV-I infection can be a critical determinant of persistent HTLV-I levels thereafter. These findings indicate that Tax-targeted vaccines may be effective for prophylaxis of ATL in a high-risk group, and also for therapy of ATL in at least half the cases. PMID- 15904465 TI - Genetic linkage between Pol iota deficiency and increased susceptibility to lung tumors in mice. AB - Pol iota is a member of the Y-family DNA polymerases, characterized by their capacity for translesion DNA synthesis and low fidelity base incorporation, and has therefore been assumed to play important roles in mutagenesis and carcinogenesis. In fact, the mouse Pol iota gene is located within the Par2 (pulmonary adenoma resistance 2) locus on distal chromosome 18, which we have identified as a major susceptibility locus regarding urethane induction of pulmonary adenomas. Indeed, Pol iota has been suggested to be a candidate for Par2 from both the genetic and biological standpoints. Taking advantage of 129X1/SvJ mice naturally deficient in Pol iota due to a nonsense mutation within the coding region of the gene, we here analyzed urethane-treated (A/J x 129X1/SvJ)F(1) x A/J backcross and (A/J x 129X1/SvJ)F(2) intercross mice and observed the defective 129X1/SvJ Pol iota allele to be genetically linked with an increased susceptibility to lung tumors relative to the A/J allele. Thus, among the already known mouse Pol iota alleles, the defective 129X1/SvJ allele is associated exclusively with the highest susceptibility to lung tumors. The result indicates a possibility that the Pol iota gene may participate in error-free repair of damaged DNA and prevention of lung tumor development. PMID- 15904466 TI - Suppression of azoxymethane-induced colon cancer development in rats by a prostaglandin E receptor EP1-selective antagonist. AB - Prostaglandin E(2) is involved in colon carcinogenesis through its binding to the PGE(2) receptor subtypes EP(1), EP(2), EP(3) and EP(4). We have demonstrated that administration of ONO-8711, an EP(1)-selective antagonist, suppresses development of AOM-induced ACF in C57BL/6 mice and F344 rats. ONO-8711 also reduced the numbers of intestinal polyps in Min mice. In the present study, we investigated the long-term effects of ONO-8711 on colon cancer development in rats treated with AOM. Male F344 rats were injected subcutaneously with AOM (15 mg/kg body weight) once a week for the first 2 weeks to develop colon cancer. Administration of 400 or 800 p.p.m. ONO-8711 in their diets for 32 weeks reduced the incidence, multiplicity and volume of colon carcinomas. The incidence of colon adenocarcinomas in AOM-treated rats was 97, 83 and 76% (P < 0.05) in the 0, 400 and 800 p.p.m. of ONO-8711 groups, respectively. The multiplicity of adenocarcinomas was also decreased significantly, being 3.31 +/- 0.33, 2.34 +/- 0.27 (P < 0.05) and 2.06 +/- 0.34 (P < 0.01) with 0, 400 and 800 p.p.m. of ONO 8711, respectively. Moreover, treatment with 800 p.p.m. ONO-8711 reduced the mean volume of adenocarcinomas to 49% (P < 0.05) of the value for the AOM treatment alone. Furthermore, the BrdU labeling index was decreased significantly in colon cancer cells by 800 p.p.m. ONO-8711. These results confirm that EP(1) is involved in colon carcinogenesis and that EP(1)-selective antagonists might be promising candidates for colon cancer chemopreventive agents. PMID- 15904467 TI - Overexpression of MUC13 is associated with intestinal-type gastric cancer. AB - Mucins are secreted or transmembrane glycoproteins that are expressed mainly in the digestive tract. This family of proteins has been the focus of much gastric cancer research as some transmembrane mucins are implicated in tumorigenesis and make attractive targets for cancer diagnosis and therapeutics. Mucins have also been utilized to classify gastric cancer by differentiating between gastric and intestinal phenotypes. Here we show that transmembrane mucin MUC13 is upregulated in gastric cancer. By quantitative real-time reverse transcription-polymerase chain reaction and immunoblot analysis, overexpression of MUC13 was verified in more than half of the samples examined. In immunohistochemical analysis, MUC13 staining was observed in 74 of 114 cases of gastric cancer (64.9%), predominantly in intestinal type (P < 0.001), and in 9 of 10 cases of intestinal metaplasia, precancerous lesions of intestinal-type gastric cancer, but not observed in normal gastric mucosa. Moreover, MUC13 staining patterns characteristic of histological type were identified: staining was on the apical side of tubular glands in intestinal type and on the cytoplasm in diffuse type. These results suggest that MUC13 is a good differentiation marker for gastrointestinal mucosa and that it may have a causal role that correlates with two distinct gastric tumorigenesis pathways. PMID- 15904468 TI - Notch1 oncoprotein antagonizes TGF-beta/Smad-mediated cell growth suppression via sequestration of coactivator p300. AB - The Notch proteins constitute a family of transmembrane receptors that play a pivotal role in cellular differentiation, proliferation and apoptosis. Although it has been recognized that excess Notch signaling is potentially tumorigenic, little is known about precise mechanisms through which dysregulated Notch signaling induces neoplastic transformation. Here we demonstrate that Notch signaling has a transcriptional cross-talk with transforming growth factor-beta (TGF-beta) signaling, which is well characterized by its antiproliferative effects. TGF-beta-mediated transcriptional responses are suppressed by constitutively active Notch1, and this inhibitory effect is canceled by introduction of transcriptional coactivator p300. We further show that this blockade of TGF-beta signaling is executed by the sequestration of p300 from Smad3. Moreover, in a human cervical carcinoma cell line, CaSki, in which Notch1 is spontaneously activated, suppression of Notch1 expression with small interfering RNA significantly restores the responsiveness to TGF-beta. Taken together, we propose that Notch oncoproteins promote cell growth and cancer development partly by suppressing the growth inhibitory effects of TGF-beta through sequestrating p300 from Smad3. PMID- 15904469 TI - Charcoal cigarette filters and lung cancer risk in Aichi Prefecture, Japan. AB - The lung cancer mortality rate has been lower in Japan than in the United States for several decades. We hypothesized that this difference is due to the Japanese preference for cigarettes with charcoal-containing filters, which efficiently absorb selected gas phase components of mainstream smoke including the carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. We analyzed a subset of smokers (396 cases and 545 controls) from a case-control study of lung cancer conducted in Aichi Prefecture, Japan. The risk associated with charcoal filters (73% of all subjects) was evaluated after adjusting for age, sex, education and smoking dose. The odds ratio (OR) associated with charcoal compared with 'plain' cigarette filters was 1.2 (95% confidence intervals [CI] 0.9, 1.6). The histologic-specific risks were similar (e.g. OR = 1.3, 95% CI 0.9, 2.1 for adenocarcinoma). The OR was 1.7 (95% CI 1.1, 2.9) in smokers who switched from 'plain' to charcoal brands. The mean daily number of cigarettes smoked in subjects who switched from 'plain' to charcoal brands was 22.5 and 23.0, respectively. The findings from this study did not indicate that charcoal filters were associated with an attenuated risk of lung cancer. As the detection of a modest benefit or risk (e.g. 10-20%) that can have significant public health impact requires large samples, the findings should be confirmed or refuted in larger studies. PMID- 15904470 TI - A tumor-specific gene therapy strategy targeting dysregulation of the VHL/HIF pathway in renal cell carcinomas. AB - Hypoxia-inducible factors, key transcription factors for hypoxia-dependent gene expression, play important roles in angiogenesis and tumor growth. The VHL protein binds to the alpha subunit of (HIF-alpha) for its oxygen-dependent degradation. VHL mutations are found frequently in sporadic RCC. Disruption of VHL results in an abnormal accumulation of HIF-alpha, leading to the upregulation of downstream genes such as the vascular endothelial growth factor gene. We constructed a luciferase reporter vector driven by hypoxia-responsive elements (5HRE/luc) and a therapeutic vector expressing a herpes simplex virus thymidine kinase gene (5HRE/tk). In the transient transfection assay using VHL-deficient 786-O cells, constitutive luciferase expression was detected under both aerobic and hypoxic conditions. In contrast, 786-O cells transfected with a wild-type VHL showed hypoxia-inducible luciferase activity. In in vitro MTS assay, 50% of growth inhibition of 786-O cells stably transfected with 5HRE/tk was achieved with exposure to 0.2 microg/mL of GCV under both aerobic and hypoxic conditions. Xenografts of the stable clone in SCID mice exhibited a marked regression on daily injections of GCV (50 mg/kg) for 10 days. In conclusion, a hypoxia responsive vector may have therapeutic potential for RCC with VHL mutations. PMID- 15904471 TI - Possible antitumor activity of 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS-106) against an established gemcitabine (dFdCyd)-resistant human pancreatic cancer cell line. AB - We established a variant of MIAPaCa-2 human pancreatic cancer cells that is resistant to 2',2'-difluorodeoxycytidine (gemcitabine, dFdCyd), MIAPaCa-2/dFdCyd, and elucidated the biochemical characteristics and mechanism of dFdCyd-resistance in these cells. We also evaluated 1-(3-C-ethynyl-beta-D-ribo pentofuranosyl)cytosine (ECyd, TAS-106, RNA polymerase inhibitor), a new anticancer ribonucleoside, for antitumor activity against the resistant cells in vitro and in vivo. MIAPaCa-2/dFdCyd cells were 2541-fold more resistant to dFdCyd than parental MIAPaCa-2 cells, and the major mechanism of the dFdCyd-resistance was found to be a decrease in the intracellular pool of dFdCyd and its active metabolites, which would result in a decrease in incorporation of dFdCyd triphosphate into DNA. This finding was confirmed by the discovery of decreased deoxycytidine kinase activity, increased cytidine deaminase and ribonucleotide reductase activity, and increased 5'-nucleotidase mRNA expression in the MIAPaCa 2/dFdCyd cells. The cytotoxicity of TAS-106 as an antitumor nucleoside analog was similar in both parental and dFdCyd-resistant cells, with IC(50) values of 6.25 and 6.27 nM, respectively, and this finding was supported by similar intracellular uptake and metabolism of TAS-106 in both cell lines. We also evaluated the in vivo antitumor activity of TAS-106 against MIAPaCa-2 and dFdCyd resistant MIAPaCa-2/dFdCyd tumors implanted into nude mice. The tumor growth inhibition rate of weekly additions of TAS-106 (7 mg/kg, iv) against parental and dFdCyd-resistant tumors was 73% and 76%, respectively, while that of dFdCyd administered twice a week (240 mg/kg, iv) was 84% and 34%, respectively. These results suggest that TAS-106 would contribute to the treatment of patients with advanced pancreatic carcinomas in whom dFdCyd-based chemotherapy has failed. PMID- 15904472 TI - Dendritic cell immunotherapy with poly(D,L-2,4-diaminobutyric acid)-mediated intratumoral delivery of the interleukin-12 gene suppresses tumor growth significantly. AB - A conventional DC-based immunotherapy has been tested clinically for treatment of patients with advanced cancer but requires modification to further improve the clinical results. In this study, we evaluated the in vivo antitumor effects of DC therapy, non-viral-mediated IL-12 gene therapy, and a combination of the two in a murine bilateral subcutaneous tumor model. DC therapy alone and IL-12 gene therapy alone suppressed tumor growth at the injected sites. However, the antitumor effect on the distant contralateral tumor was insufficient. When DC therapy and IL-12 gene therapy were carried out simultaneously, tumor growth was significantly suppressed bilaterally (P < 0.001). Cytolytic activity was augmented significantly in mice given the combination treatment compared to in mice treated with either DC or IL-12 gene therapy alone (P < 0.05). Microvessel density of both tumors was significantly lower in mice subjected to the combination therapy than in mice treated otherwise (P < 0.05). Furthermore, no side-effects were observed in the treated mice. DC therapy combined with non viral-mediated intratumoral IL-12 gene delivery has a synergistic antitumor effect not only on targeted tumors but also on contralateral distant tumors and may be of great potential as a therapeutic treatment for patients with advanced cancer. PMID- 15904473 TI - Systematic review and meta-analysis: proton pump inhibitor vs. ranitidine bismuth citrate plus two antibiotics in Helicobacter pylori eradication. AB - OBJECTIVE: To systematically review the Helicobacter pylori eradication efficacy with ranitidine bismuth citrate (RBC) and two antibiotics, and to conduct a meta analysis of randomized clinical trials comparing the efficacy of proton pump inhibitor (PPI) vs. RBC with two antibiotics for 1 week. METHODS: SELECTION OF STUDIES: Studies evaluating RBC plus two antibiotics were considered. For the meta-analysis, randomized controlled trials comparing PPI vs. RBC plus two antibiotics for 1 week were included. SEARCH STRATEGY: Electronic and manual bibliographical searches. Assessment of study quality and data extraction: Independently done by two reviewers. DATA SYNTHESIS: 'Intention-to-treat' eradication rate. Meta-analysis was performed, combining the odds ratios (OR) of the individual studies. Subanalysis: Depending on the type of antibiotics and the quality of the studies. RESULTS: Mean H. pylori eradication with 7-day RBC clarithromycin-amoxicillin, RBC-clarithromycin-nitroimidazole, and RBC amoxicillin-nitroimidazole was 83%, 86%, and 71%, respectively. The meta-analysis showed comparable efficacy with RBC and PPI when they were combined with clarithromycin and amoxicillin (OR = 1.11; 95% CI = 0.88-1.40), or with amoxicillin and metronidazole (OR = 0.92; 95%CI = 0.60-1.41). However, when comparing PPI vs. RBC plus clarithromycin and a nitroimidazole, higher cure rates with RBC than with PPI were demonstrated (OR = 1.65; 95% CI = 1.15-2.37). CONCLUSION: The efficacy of RBC and PPI-based triple regimens were comparable when using the clarithromycin-amoxicillin or the amoxicillin-metronidazole combination. However, RBC seems to have a higher efficacy than PPI when clarithromycin and a nitroimidazole are the antibiotics prescribed. Therefore, if one prefers to use the clarithromycin-nitroimidazole regimen, RBC should be used instead of a PPI. PMID- 15904474 TI - Interleukin-2 gene polymorphisms associated with increased risk of gastric atrophy from Helicobacter pylori infection. AB - BACKGROUND: Gastric atrophy induced by Helicobacter pylori is thought to predispose patients to noncardiac gastric cancer development. However, the host genetic factors that influence the progression of gastric atrophy have not been elucidated. In this study, we examined the effects of cytokine polymorphisms on H. pylori-induced gastric atrophy. METHODS: Blood samples were taken from 454 Japanese subjects. The interleukin-2 (IL-2; T-330G), IL-4 (C-33T), and IL-13 (C 1111T) polymorphisms were genotyped by polymerase chain reaction with confronting two-pair primers (PCR-CTPP). Anti-H. pylori IgG antibody and pepsinogen I and II were measured to diagnose H. pylori infection and atrophic gastritis. RESULTS: The odds ratios (ORs) for the association between IL-2 polymorphism [OR = 2.78, 95% CI (confidence interval) = 1.26-6.17 (T/T to G/G)] or IL-4 polymorphism [OR = 2.22, 95% CI = 1.01-4.89 (T/C to C/C)] were increased significantly with gastric atrophy, whereas the corresponding OR of IL-13 polymorphism was decreased with gastric atrophy [OR = 0.61, 95% CI = 0.39-0.96 (C/T and T/T to C/C)]. There were no significant H. pylori seropositivity-related differences between these polymorphisms. We examined the relationship between these polymorphisms and gastric atrophy separately in H. pylori-seropositive and -seronegative groups. In the H. pylori-seropositive group, the IL-2 T/T (OR = 2.78, 95% CI = 1.12-6.93) had a significant association with gastric atrophy. CONCLUSIONS: These results reveal that the IL-2 gene polymorphism is associated with an increased risk of gastric atrophy induced by H. pylori infection and might predispose to gastric cancer. PMID- 15904475 TI - Effect of Helicobacter pylori infection on the expression of DNA mismatch repair protein. AB - BACKGROUND: Helicobacer pylori infection is a major gastric cancer risk factor. Deficient DNA mismatch repair (MMR) caused by H. pylori may underlie microsatellite instability (MSI) in the gastric epithelium and may represent a major mechanism of mutation accumulation in the gastric mucosa during the early stages of H. pylori-associated gastric carcinogenesis. In this study, we examined the expression of DNA MMR protein (hMLH1 and hMSH2) in patients with chronic H. pylori infection before and after eradication of the infection. MATERIALS AND METHODS: Gastric tissue samples were collected from 60 patients with H. pylori gastritis and peptic ulcer disease before and after eradication of the infection. The DNA MMR protein expression (hMLH1 and hMSH2) was determined by immunohistochemical staining in 60 patients before and after H. pylori eradication. The percentage of epithelial cell nuclei and intensity of staining were then compared in gastric biopsies before and after eradication. RESULTS: The percentage of hMLH1 (76.60 +/- 20.27, 84.82 +/- 12.73, p=.01) and hMSH2 (82.36 +/ 12.86, 88.11 +/- 9.27, p<.05) positive epithelial cells significantly increased in 53 patients who became H. pylori-negative after eradication therapy. However, the intensity of hMLH1 and hMSH2 staining was not significantly different. In those 7 patients, who did not respond to the eradication therapy and were still H. pylori-positive, the percent positivity and intensity of hMLH1 and hMSH2 staining did not change. CONCLUSIONS: The expression of DNA MMR proteins increased in the gastric mucosa after H. pylori eradication, indicating that H. pylori gastritis may be associated with a reduced DNA MMR system during infection. The effect of H. pylori infection on MMR protein expression appears to be at least partially reversible after H. pylori eradication. These data suggest that H. pylori gastritis might lead to a deficiency of DNA MMR in gastric epithelium that may increase the risk of mutation accumulation in the gastric mucosa cells during chronic H. pylori infection. PMID- 15904476 TI - Evaluation of the homologous recombination in Helicobacter pylori. AB - BACKGROUND: Most strategies for direct mutagenesis of Helicobacter pylori primarily involve genomic DNA cloning which is a time-consuming and expensive technique. METHODS: To make a gene replacement, we propose a strategy using polymerase chain reaction (PCR) amplicons to allow a double homologous recombination in the genome of H. pylori. Different strains were used to validate this strategy and we describe how the amplicon insertion was made with accuracy. Moreover, we looked for the shortest homologous sequence needed to allow a specific gene replacement in H. pylori without any deletion, insertion or mutation at the recombination site. All of the experiments were performed at the flaA locus, whose gene encodes the major flagellin. RESULTS: Amplicons bearing 500 or 150 bp flanking regions of flaA on each side (depending on the strain) were sufficient to allow the specific insertion of a 1173 bp chloramphenicol cassette into the genome of H. pylori. The insertion was accurate with no substitutions at the insertion locus. CONCLUSIONS: This information opens the door to other strategies for mutagenesis used for the identification of virulence factors without deleting genes, which would not be based on a negative screening system. For example, they could be useful in performing protein fusion for a better understanding of the virulence factor's mechanism. PMID- 15904477 TI - Toll-like receptor 2-mediated gene expression in epithelial cells during Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori is the major pathogen causing chronic gastritis and peptic ulcer disease and is closely linked to gastric malignancy. We have previously shown that H. pylori-induced NF-(kappa)B activation and interleukin (IL)-8 secretion are mediated by Toll-like receptor (TLR) 2 in epithelial cells. However, the TLR2-mediated global gene expression profile of the epithelial cell during H. pylori infection is still unknown. The goal of this study was to identify TLR2-regulated genes in epithelial cells induced by H. pylori. MATERIALS AND METHODS: The HEK293 and HEK-TLR2 cells were cocultured with H. pylori 26695 for 6 hours. Total RNA was extracted and hybridized to the Affymetrix human U133A microarray chipset, which contains 22,283 total probe sets including 14,285 genes. Data analyses were performed using affymetrix suite 5 software. The expression of selected genes in gastric epithelial cells AGS and MKN45 was monitored by quantitative reverse transcriptase-polymerase chain reaction (RT PCR). RESULTS: Forty-six genes, contained in 57 probe sets, were induced > 2-fold and three genes (five probe sets) decreased > 2-fold by H. pylori infection of HEK293 cells. Fifty-four genes, contained in 69 probe sets, were induced > 2 fold, whereas only 1 gene was repressed > 2-fold in H. pylori-infected HEK-TLR2 cells. Comparisons of genes induced in HEK293 or HEK-TLR2 cells identified 28 genes whose expression was dependent on the presence of TLR2. Seventeen genes were selected and their expression was assessed using the quantitative RT-PCR in gastric epithelial cells during H. pylori infection. Eight of the 17 genes showed distinct expression patterns in AGS and MKN45 cells after H. pylori stimulation. CONCLUSIONS: The current study investigated the TLR2-mediated global gene changes after H. pylori stimulation in the epithelial cell system. This approach will be helpful in identifying genes whose expression is mediated by specific TLRs and in determining the cellular responses that are responsible for diverse signal pathways during H. pylori infection. PMID- 15904478 TI - Polymerase chain reaction--restriction fragment length polymorphism analysis of clarithromycin-resistant Helicobacter pylori infection in children using stool sample. AB - BACKGROUND: To analyze clarithromycin-resistant Helicobacter pylori infection in children, we developed a method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using stool samples. MATERIALS AND METHODS: Twenty-three children without significant upper abdominal symptoms were included (mean age 7.0 years). Of these, 18 and five were diagnosed as H. pylori positive and -negative, respectively, by the H. pylori stool antigen test (HpSA). The DNA from the stool samples was purified using the QIAamp DNA Stool Minikit (QIAGEN). The PCR was performed on the purified DNA using oligonucleotide primers designed to amplify the 23S rRNA gene of H. pylori. The PCR products were reacted with restriction enzymes MboII, BceAI, and BsaI to detect mutations A2142G, A2142C, and A2143G, respectively. RESULTS: Sixteen of the 18 HpSA-positive samples were PCR-positive, and all five HpSA-negative samples were PCR-negative. Thus, the PCR had 89% sensitivity and 100% specificity, with 91% accuracy in reference to HpSA. Of the 16 PCR-positive samples, one and four were digested with MboII and BsaI, respectively, indicating 31% prevalence of CAM-resistance. CONCLUSIONS: We conclude that the PCR-RFLP using stool samples is a rapid and reliable method to noninvasively detect clarithromycin-resistant H. pylori infection in children. It may be useful before choosing regimens of H. pylori eradication. PMID- 15904479 TI - Increase of antigen-presenting cells in the gastric mucosa of Helicobacter pylori infected children. AB - BACKGROUND: Infection with Helicobacter pylori leads to an increase of T cells in the gastric mucosa of children. In contrast to peripheral blood, where monocytes are the most abundant antigen-presenting cells, CD14+ macrophages are very rare in infected gastric mucosa. We postulated that other types of antigen-presenting cells must be present in infected gastric mucosa. MATERIAL AND METHODS: Antral biopsies were obtained from 56 children. The cellular expression of major histocompatibility complex (MHC) class II molecules, CD1a/b, and CD23, which are involved in antigen presentation were analyzed by immunohistochemistry. In addition, T cells (CD4, CD8, CD25, and gamma/delta-TCR), B cells (anti-IgM), macrophages (CD14) and granulocytes (CD15) were quantified. RESULTS: Twenty-eight children were H. pylori-infected. Thirteen children were healthy, 15 had other gastric pathologies. T cells (p<.0001), B cells (p<.0001), CD23+ (p<.0001), and CD1a/b+ (p<.005) cells were significantly increased in the lamina propria of H. pylori-infected children, whereas macrophages were rare without significant differences among the groups. Within the epithelium, CD8+ T lymphocytes predominated clearly over CD4+ cells. H. pylori-negative children had only few MHC class II-positive cells within the gastric epithelium, whereas MHC class II antigens were strongly expressed on epithelial cells (p<.0001) of all H. pylori infected children. CONCLUSION: Helicobacter pylori infection leads to an enhanced expression of antigen-presenting molecules together with a parallel rise of T cells in the lamina propria. This may represent an effort of the immune system to optimize local immune responses against H. pylori. We speculate that the epithelium participates in the initiation of a local immune response against H. pylori. PMID- 15904480 TI - Natural colonization with Helicobacter species and the development of inflammatory bowel disease in interleukin-10-deficient mice. AB - BACKGROUND: The interleukin-10-deficient (IL-10-/-) mice maintained in specific pathogen-free (SPF) conditions develop typhlocolitis when experimentally infected with Helicobacter species. However, there is limited information regarding the role of Helicobacter species that naturally colonize IL-10-/- mice in typhlocolitis development. The aim of this study was to examine in SPF IL-10-/- mice the association between natural colonization specific Helicobacter species and typhlocolitis development. MATERIAL AND METHODS: Cecum and proximal colon from 72 C57BL/6 x 129/Ola IL-10-/- mice (8-20 weeks old) were removed for DNA extraction and histologic evaluation. Genus-specific polymerase chain reaction- denaturing gradient gel electrophoresis (PCR-DGGE) and species-specific PCR were used to detect Helicobacter species. Mice were grouped by age, sex, and Helicobacter colonization status, and their histologic scores were compared. The development of clinical typhlocolitis was observed in a further 12 mice. RESULTS: Species-specific PCR showed that mice were colonized with Helicobacter ganmani and/or Helicobacter hepaticus. The PCR-DGGE detected H. ganmani, H. hepaticus and an H. ganmani-like organism. The histologic scores in mice colonized with H. hepaticus were significantly higher than that in mice colonized with H. ganmani. Male mice showed significantly higher histologic scores than female mice. Four of the 12 mice developed clinical typhlocolitis in 38 weeks. CONCLUSION: Natural colonization with different Helicobacter species was found in IL-10-/- mice within the same breeding colony. The severity of typhlocolitis differed according to the colonizing Helicobacter species. Furthermore, the rate of typhlocolitis development in IL-10-/- mice naturally colonized with Helicobacter species was significantly slower than that reported in experimentally infected mice. PMID- 15904481 TI - Inhibitory effects of polyphenols on gastric injury by Helicobacter pylori VacA toxin. AB - BACKGROUND: Helicobacter pylori induces gastric damage and may be involved in the pathogenesis of gastric cancer. H. pylori-vacuolating cytotoxin, VacA, is one of the important virulence factors, and is responsible for H. pylori-induced gastritis and ulceration. The aim of this study is to assess whether several naturally occurring polyphenols inhibit VacA activities in vitro and in vivo. MATERIALS AND METHODS: Effects of polyphenols on VacA were quantified by the inhibition of: 1, vacuolation; 2, VacA binding to AZ-521 or G401 cells or its receptors; 3, VacA internalization. Effects of hop bract extract (HBT) containing high molecular weight polymerized catechin on VacA in vivo were investigated by quantifying gastric damage after oral administration of toxins to mice. RESULTS: HBT had the strongest inhibitory activity among the polyphenols investigated. HBT inhibited, in a concentration-dependent manner: 1, VacA binding to its receptors, RPTP(alpha) and RPTP(beta); 2, VacA uptake; 3, VacA-induced vacuolation in susceptible cells. In addition, oral administration of HBT with VacA to mice reduced VacA-induced gastric damage at 48 hours. In vitro, VacA formed a complex with HBT. CONCLUSIONS: HBT may suppress the development of inflammation and ulceration caused by H. pylori VacA, suggesting that HBT may be useful as a new type of therapeutic agent for the prevention of gastric ulcer and inflammation caused by VacA. PMID- 15904482 TI - Isogenic variation of Helicobacter pylori strain resulting in heteroresistant antibacterial phenotypes in a single host in vivo. AB - BACKGROUND: Antibiotic-susceptible and -resistant Helicobacter pylori can be present simultaneously in the same host. The aim of this study was to evaluate the genomic diversity of H. pylori strains resulting in heteroresistant antibacterial phenotypes. MATERIALS AND METHODS: Twenty-one pairs of H. pylori strains isolated from the antrum and body displaying heteroresistant antibacterial phenotypes were included. We compared the genotypes of paired isolates by random arbitrarily primed polymerase chain reaction (PCR), flagella gene PCR-based restriction fragment length polymorphism, and flaA gene sequencing. In metronidazole-heteroresistant isolates, the sequence variation of rdxA and frxA genes was analyzed using phylogenetic analysis. RESULTS: The DNA fingerprinting patterns of the paired isolates revealed that 12 pairs (57.1%) were identical, whereas one pair (3.8%) was different. The remaining eight pairs (38.1%) of isolates showed minor heterogenecity in fingerprinting patterns. In flaA gene sequencing, these identical and similar isolates showed close sequence similarity between the antrum and body, whereas different isolate showed 31 points of different nucleotide sequences. Phylogenetic analysis of the metronidazole-heteroresistant pairs showed consistent genetic relatedness of each paired isolates despite the sequence variation of the rdxA or frxA genes in five pairs (71.4%). CONCLUSIONS: These results suggest that continuing genomic diversities in the same strain may play an important role in modulating the antibiotic-heteroresistant H. pylori in vivo. PMID- 15904483 TI - Children of Helicobacter pylori-infected dyspeptic mothers are predisposed to H. pylori acquisition with subsequent iron deficiency and growth retardation. AB - BACKGROUND: We tested whether Helicobacter pylori-infected dyspeptic mothers had a higher rate of H. pylori infection in their children, and whether such H. pylori-infected children were predisposed to iron deficiency or growth retardation. MATERIALS AND METHODS: A total of 163 children from 106 dyspeptic mothers (58 with and 48 without H. pylori infection) were enrolled to evaluate body weight, height, hemoglobin, serum ferritin, and H. pylori infection using the 13C-urea breath test. A questionnaire was used to evaluate demographic factors of each child. RESULTS: The rate of H. pylori infection in children with H. pylori-infected dyspeptic mothers was higher than that of children with noninfected mothers (20.5% vs. 5.3%; p<.01, OR: 4.6, 95% CI: 1.5-14.2). The rate of H. pylori infection in children elevated as the number of their H. pylori infected siblings increased (p<.01). For children below 10 years of age, H. pylori infection was closely related to low serum ferritin and body weight growth (p<.05). CONCLUSION: The children of H. pylori-infected dyspeptic mothers had an increased risk for such infection. The risk further increased once their siblings were infected. H. pylori infection in pre-adolescent children may determine iron deficiency and growth retardation. PMID- 15904484 TI - Gastric acidity in patients with follicular gastritis is significantly reduced, but can be normalized after eradication for Helicobacter pylori. AB - BACKGROUND: Follicular gastritis is thought to be caused by Helicobacter pylori infection. However, the pathophysiology of it remains unclear. MATERIALS AND METHODS: We assessed gastric acidity in 15 patients with follicular gastritis, aged 20-37 years, using a 24-hour intragastric pH-metry, as well as by histologic and serologic evaluations; and compared it with that in other age-matched groups: 18 cases of H. pylori-positive antrum-predominant gastritis, 12 of pangastritis, and 24 H. pylori-negative normals. In eight cases with follicular gastritis, it was re-assessed 6 months after the eradication therapy for H. pylori. RESULTS: During nighttime, the percentage of time with intragastric pH above 3.0 in follicular gastritis was significantly higher than that in normals (p<.0001), and in antrum-predominant gastritis (p<.001), but was comparable with that in pangastritis. In the daytime period, this parameter in follicular gastritis was significantly higher than that in normal (p<.001), in antrum-predominant gastritis (p<.001), and in pangastritis (p<.05). Marked mononuclear cell and neutrophil infiltration but no apparent glandular atrophy were observed in both the antrum and corpus. Serum pepsinogen I/II ratio was significantly lower in follicular gastritis than that in normals (p<.0001) and in antrum-predominant gastritis (p<.001), whereas serum gastrin was significantly higher than that in normals (p<.0001), in antrum-predominant gastritis (p<.01) and in pangastritis (p<.05). After eradication for H. pylori, all of the parameters in follicular gastritis were altered to the same ranges as those in normals. CONCLUSIONS: In follicular gastritis, gastric acidity is significantly reduced, but can be normalized by eradication of H. pylori. It can thus be speculated that inflammatory cytokines or H. pylori-infection-induced prostaglandins might strongly inhibit gastric acid secretion in follicular gastritis. PMID- 15904485 TI - The effect of refurbishing a UK steel plant on PM10 metal composition and ability to induce inflammation. AB - BACKGROUND: In the year 2000 Corus closed its steel plant operations in Redcar, NE of England temporarily for refurbishment of its blast furnace. This study investigates the impact of the closure on the chemical composition and biological activity of PM10 collected in the vicinity of the steel plant. METHODS: The metal content of PM10 samples collected before during and after the closure was measured by ICP-MS in order to ascertain whether there was any significant alteration in PM10 composition during the steel plant closure. Biological activity was assessed by instillation of 24 hr PM10 samples into male Wistar rats for 18 hr (n = 6). Inflammation was identified by the cellular and biochemical profile of the bronchoalveolar lavage fluid. Metal chelation of PM10 samples was conducted using Chelex beads prior to treatment of macrophage cell line, J774, in vitro and assessment of pro-inflammatory cytokine expression. RESULTS: The total metal content of PM10 collected before and during the closure period were similar, but on reopening of the steel plant there was a significant 3-fold increase (p < 0.05) compared with the closure and pre-closure samples. Wind direction prior to the closure was predominantly from the north, compared to south westerly during the closure and re-opened periods. Of metals analysed, iron was most abundant in the total and acid extract, while zinc was the most prevalent metal in the water-soluble fraction. Elevated markers of inflammation included a significant increase (p < 0.01) in neutrophil cell numbers in the bronchoalveolar lavage of rats instilled with PM10 collected during the reopened period, as well as significant increases in albumin (p < 0.05). Extracts of PM10 from the pre-closure and closure periods did not induce any significant alterations in inflammation or lung damage. The soluble and insoluble extractable PM10 components washed from the reopened period both induced a significant increase in neutrophil cell number (p < 0.05) when compared to the control, and these increases when added together approximately equalled the inflammation induced by the whole sample. PM10 from the re-opened period stimulated J774 macrophages to generate TNF-alpha protein and this was significantly prevented by chelating the metal content of the PM10 prior to addition to the cells. CONCLUSION: PM10-induced inflammation in the rat lung was related to the concentration of metals in the PM10 samples tested, and activity was found in both the soluble and insoluble fractions of the particulate pollutant. PMID- 15904486 TI - Pattern similarity study of functional sites in protein sequences: lysozymes and cystatins. AB - BACKGROUND: Although it is generally agreed that topography is more conserved than sequences, proteins sharing the same fold can have different functions, while there are protein families with low sequence similarity. An alternative method for profile analysis of characteristic conserved positions of the motifs within the 3D structures may be needed for functional annotation of protein sequences. Using the approach of quantitative structure-activity relationships (QSAR), we have proposed a new algorithm for postulating functional mechanisms on the basis of pattern similarity and average of property values of side-chains in segments within sequences. This approach was used to search for functional sites of proteins belonging to the lysozyme and cystatin families. RESULTS: Hydrophobicity and beta-turn propensity of reference segments with 3-7 residues were used for the homology similarity search (HSS) for active sites. Hydrogen bonding was used as the side-chain property for searching the binding sites of lysozymes. The profiles of similarity constants and average values of these parameters as functions of their positions in the sequences could identify both active and substrate binding sites of the lysozyme of Streptomyces coelicolor, which has been reported as a new fold enzyme (Cellosyl). The same approach was successfully applied to cystatins, especially for postulating the mechanisms of amyloidosis of human cystatin C as well as human lysozyme. CONCLUSION: Pattern similarity and average index values of structure-related properties of side chains in short segments of three residues or longer were, for the first time, successfully applied for predicting functional sites in sequences. This new approach may be applicable to studying functional sites in un-annotated proteins, for which complete 3D structures are not yet available. PMID- 15904487 TI - Are scale-free networks robust to measurement errors? AB - BACKGROUND: Many complex random networks have been found to be scale-free. Existing literature on scale-free networks has rarely considered potential false positive and false negative links in the observed networks, especially in biological networks inferred from high-throughput experiments. Therefore, it is important to study the impact of these measurement errors on the topology of the observed networks. RESULTS: This article addresses the impact of erroneous links on network topological inference and explores possible error mechanisms for scale free networks with an emphasis on Saccharomyces cerevisiae protein interaction networks. We study this issue by both theoretical derivations and simulations. We show that the ignorance of erroneous links in network analysis may lead to biased estimates of the scale parameter and recommend robust estimators in such scenarios. Possible error mechanisms of yeast protein interaction networks are explored by comparisons between real data and simulated data. CONCLUSION: Our studies show that, in the presence of erroneous links, the connectivity distribution of scale-free networks is still scale-free for the middle range connectivities, but can be greatly distorted for low and high connecitivities. It is more appropriate to use robust estimators such as the least trimmed mean squares estimator to estimate the scale parameter gamma under such circumstances. Moreover, we show by simulation studies that the scale-free property is robust to some error mechanisms but untenable to others. The simulation results also suggest that different error mechanisms may be operating in the yeast protein interaction networks produced from different data sources. In the MIPS gold standard protein interaction data, there appears to be a high rate of false negative links, and the false negative and false positive rates are more or less constant across proteins with different connectivities. However, the error mechanism of yeast two-hybrid data may be very different, where the overall false negative rate is low and the false negative rates tend to be higher for links involving proteins with more interacting partners. PMID- 15904488 TI - The effects of normalization on the correlation structure of microarray data. AB - BACKGROUND: Stochastic dependence between gene expression levels in microarray data is of critical importance for the methods of statistical inference that resort to pooling test-statistics across genes. It is frequently assumed that dependence between genes (or tests) is sufficiently weak to justify the proposed methods of testing for differentially expressed genes. A potential impact of between-gene correlations on the performance of such methods has yet to be explored. RESULTS: The paper presents a systematic study of correlation between the t-statistics associated with different genes. We report the effects of four different normalization methods using a large set of microarray data on childhood leukemia in addition to several sets of simulated data. Our findings help decipher the correlation structure of microarray data before and after the application of normalization procedures. CONCLUSION: A long-range correlation in microarray data manifests itself in thousands of genes that are heavily correlated with a given gene in terms of the associated t-statistics. By using normalization methods it is possible to significantly reduce correlation between the t-statistics computed for different genes. Normalization procedures affect both the true correlation, stemming from gene interactions, and the spurious correlation induced by random noise. When analyzing real world biological data sets, normalization procedures are unable to completely remove correlation between the test statistics. The long-range correlation structure also persists in normalized data. PMID- 15904489 TI - A multistep bioinformatic approach detects putative regulatory elements in gene promoters. AB - BACKGROUND: Searching for approximate patterns in large promoter sequences frequently produces an exceedingly high numbers of results. Our aim was to exploit biological knowledge for definition of a sheltered search space and of appropriate search parameters, in order to develop a method for identification of a tractable number of sequence motifs. RESULTS: Novel software (COOP) was developed for extraction of sequence motifs, based on clustering of exact or approximate patterns according to the frequency of their overlapping occurrences. Genomic sequences of 1 Kb upstream of 91 genes differentially expressed and/or encoding proteins with relevant function in adult human retina were analyzed. Methodology and results were tested by analysing 1,000 groups of putatively unrelated sequences, randomly selected among 17,156 human gene promoters. When applied to a sample of human promoters, the method identified 279 putative motifs frequently occurring in retina promoters sequences. Most of them are localized in the proximal portion of promoters, less variable in central region than in lateral regions and similar to known regulatory sequences. COOP software and reference manual are freely available upon request to the Authors. CONCLUSION: The approach described in this paper seems effective for identifying a tractable number of sequence motifs with putative regulatory role. PMID- 15904490 TI - Involvement of CD147 in overexpression of MMP-2 and MMP-9 and enhancement of invasive potential of PMA-differentiated THP-1. AB - BACKGROUND: During infection and inflammation, circulating blood monocytes migrate from the intravascular compartments to the extravascular compartments, where they mature into tissue macrophages. The maturation process prepares the cells to actively participate in the inflammatory and immune responses, and many factors have been reported to be involved in the process. We found in our study that CD147 played a very important role in this process. RESULTS: By using PMA differentiated human monocyte cells line THP-1, we found that CD147 mediated matrix metalloproteinases (MMPs) expression of the leukemic THP-1 cells and thus enhanced the invasiveness of THP-1 cells. After 24 hours of PMA-induced monocyte differentiation, the mean fluorescence intensity of CD147 in differentiated THP-1 cells (289.61 +/- 31.63) was higher than that of the undifferentiated THP-1 cells (205.1 +/- 19.25). There was a significant increase of the levels of proMMP-2, proMMP-9 and their activated forms in the differentiated THP-1 cells. Invasion assays using reconstituted basement membrane showed a good correlation between the invasiveness of THP-1 cells and the production of MMP-2 and MMP-9. The difference in the MMPs expression and the invasive ability was significantly blocked by HAb18G/CD147 antagonistic peptide AP-9. The inhibitory rate of the secretion of proMMP-9 in the undifferentiated THP-1 cells was 45.07%. The inhibitory rate of the secretion of proMMP-9, the activated MMP-9 and proMMP-2 in the differentiated THP-1 cells was 52.90%, 53.79% and 47.80%, respectively. The inhibitory rate of invasive potential in the undifferentiated cells and the differentiated THP-1 cells was 41.82 % and 25.15%, respectively. CONCLUSION: The results suggest that the expression of CD147 is upregulated during the differentiation of monocyte THP-1 cells to macrophage cells, and CD147 induces the secretion and activation of MMP-2 and MMP-9 and enhances the invasive ability of THP-1 cells. The matured monocytes / macrophages, via their high expression of CD147, may play an important role in promoting the tissue repair or tissue damage during their inflammatory response. PMID- 15904491 TI - The complexity of selection at the major primate beta-defensin locus. AB - BACKGROUND: We have examined the evolution of the genes at the major human beta defensin locus and the orthologous loci in a range of other primates and mouse. For the first time these data allow us to examine selective episodes in the more recent evolutionary history of this locus as well as the ancient past. We have used a combination of maximum likelihood based tests and a maximum parsimony based sliding window approach to give a detailed view of the varying modes of selection operating at this locus. RESULTS: We provide evidence for strong positive selection soon after the duplication of these genes within an ancestral mammalian genome. Consequently variable selective pressures have acted on beta defensin genes in different evolutionary lineages, with episodes both of negative, and more rarely positive selection, during the divergence of primates. Positive selection appears to have been more common in the rodent lineage, accompanying the birth of novel, rodent-specific beta-defensin genes. These observations allow a fuller understanding of the evolution of mammalian innate immunity. In both the rodent and primate lineages, sites in the second exon have been subject to positive selection and by implication are important in functional diversity. A small number of sites in the mature human peptides were found to have undergone repeated episodes of selection in different primate lineages. Particular sites were consistently implicated by multiple methods at positions throughout the mature peptides. These sites are clustered at positions predicted to be important for the specificity of the antimicrobial or chemoattractant properties of beta-defensins. Surprisingly, sites within the prepropeptide region were also implicated as being subject to significant positive selection, suggesting previously unappreciated functional significance for this region. CONCLUSIONS: Identification of these putatively functional sites has important implications for our understanding of beta-defensin function and for novel antibiotic design. PMID- 15904492 TI - On the use of haplotype phylogeny to detect disease susceptibility loci. AB - BACKGROUND: The cladistic approach proposed by Templeton has been presented as promising for the study of the genetic factors involved in common diseases. This approach allows the joint study of multiple markers within a gene by considering haplotypes and grouping them in nested clades. The idea is to search for clades with an excess of cases as compared to the whole sample and to identify the mutations defining these clades as potential candidate disease susceptibility sites. However, the performance of this approach for the study of the genetic factors involved in complex diseases has never been studied. RESULTS: In this paper, we propose a new method to perform such a cladistic analysis and we estimate its power through simulations. We show that under models where the susceptibility to the disease is caused by a single genetic variant, the cladistic test is neither really more powerful to detect an association nor really more efficient to localize the susceptibility site than an individual SNP testing. However, when two interacting sites are responsible for the disease, the cladistic analysis greatly improves the probability to find the two susceptibility sites. The impact of the linkage disequilibrium and of the tree characteristics on the efficiency of the cladistic analysis are also discussed. An application on a real data set concerning the CARD15 gene and Crohn disease shows that the method can successfully identify the three variant sites that are involved in the disease susceptibility. CONCLUSION: The use of phylogenies to group haplotypes is especially interesting to pinpoint the sites that are likely to be involved in disease susceptibility among the different markers identified within a gene. PMID- 15904493 TI - Software for optimization of SNP and PCR-RFLP genotyping to discriminate many genomes with the fewest assays. AB - BACKGROUND: Microbial forensics is important in tracking the source of a pathogen, whether the disease is a naturally occurring outbreak or part of a criminal investigation. RESULTS: A method and SPR Opt (SNP and PCR-RFLP Optimization) software to perform a comprehensive, whole-genome analysis to forensically discriminate multiple sequences is presented. Tools for the optimization of forensic typing using Single Nucleotide Polymorphism (SNP) and PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) analyses across multiple isolate sequences of a species are described. The PCR-RFLP analysis includes prediction and selection of optimal primers and restriction enzymes to enable maximum isolate discrimination based on sequence information. SPR Opt calculates all SNP or PCR-RFLP variations present in the sequences, groups them into haplotypes according to their co-segregation across those sequences, and performs combinatoric analyses to determine which sets of haplotypes provide maximal discrimination among all the input sequences. Those set combinations requiring that membership in the fewest haplotypes be queried (i.e. the fewest assays be performed) are found. These analyses highlight variable regions based on existing sequence data. These markers may be heterogeneous among unsequenced isolates as well, and thus may be useful for characterizing the relationships among unsequenced as well as sequenced isolates. The predictions are multi-locus. Analyses of mumps and SARS viruses are summarized. Phylogenetic trees created based on SNPs, PCR-RFLPs, and full genomes are compared for SARS virus, illustrating that purported phylogenies based only on SNP or PCR-RFLP variations do not match those based on multiple sequence alignment of the full genomes. CONCLUSION: This is the first software to optimize the selection of forensic markers to maximize information gained from the fewest assays, accepting whole or partial genome sequence data as input. As more sequence data becomes available for multiple strains and isolates of a species, automated, computational approaches such as those described here will be essential to make sense of large amounts of information, and to guide and optimize efforts in the laboratory. The software and source code for SPR Opt is publicly available and free for non profit use at http://www.llnl.gov/IPandC/technology/software/softwaretitles/spropt.php. PMID- 15904494 TI - Role of core promoter sequences in the mechanism of swarmer cell-specific silencing of gyrB transcription in Caulobacter crescentus. AB - BACKGROUND: Each Caulobacter crescentus cell division yields two distinct cell types: a flagellated swarmer cell and a non-motile stalked cell. The swarmer cell is further distinguished from the stalked cell by an inability to reinitiate DNA replication, by the physical properties of its nucleoid, and its discrete program of gene expression. Specifically, with regard to the latter feature, many of the genes involved in DNA replication are not transcribed in swarmer cells. RESULTS: We show that for one of these genes involved in DNA replication, gyrB, its pattern of temporal expression depends upon an 80 base pair promoter region with strong resemblance to the Caulobacter crescentus sigma73 consensus promoter sequence; regulation does not appear to be affected by the general strength of the promoter activity, as mutations that increased its conformity with the consensus did not affect its cell-cycle expression pattern. Transcription from the gyrB promoter in vitro required only the presence of the sigma73 RNA polymerase (from E. coli) and the requisite nucleoside triphosphates, although a distinct binding activity, present in crude whole-cell extracts, formed a complex gyrB promoter DNA. We also assayed the effect on gyrB expression in strains containing mutations in either smc or dps, two genes encoding proteins that condense DNA. However we found there was no change in the temporal pattern of gyrB transcription in strains containing deletions in either of these genes. CONCLUSION: These experiments demonstrate that gyrB transcription does not require any auxiliary factors, suggesting that temporal regulation is not dependent upon an activator protein. Swarmer-specific silencing may not be attributable to the observed physical difference in the swarmer cell nucleoid, since mutations in either smc or dps, two genes encoding proteins that condense DNA, did not alter the temporal pattern of gyrB transcription in strains containing deletions in either of these genes. Rather a repressor that specifically recognizes sequences in the gyrB promoter region that are also probably essential for transcription, is likely to be responsible for controlling cell cycle expression. PMID- 15904495 TI - Selecting representative model micro-organisms. AB - BACKGROUND: Micro-biological research relies on the use of model organisms that act as representatives of their species or subspecies, these are frequently well characterized laboratory strains. However, it has often become apparent that the model strain initially chosen does not represent important features of the species. For micro-organisms, the diversity of their genomes is such that even the best possible choice of initial strain for sequencing may not assure that the genome obtained adequately represents the species. To acquire information about a species' genome as efficiently as possible, we require a method to choose strains for analysis on the basis of how well they represent the species. RESULTS: We develop the Best Total Coverage (BTC) method for selecting one or more representative model organisms from a group of interest, given that rough genetic distances between the members of the group are known. Software implementing a "greedy" version of the method can be used with large data sets, its effectiveness is tested using both constructed and biological data sets. CONCLUSION: In both the simulated and biological examples the greedy-BTC method outperformed random selection of model organisms, and for two biological examples it outperformed selection of model strains based on phylogenetic structure. Although the method was designed with microbial species in mind, and is tested here on three microbial data sets, it will also be applicable to other types of organism. PMID- 15904496 TI - HinT proteins and their putative interaction partners in Mollicutes and Chlamydiaceae. AB - BACKGROUND: HinT proteins are found in prokaryotes and eukaryotes and belong to the superfamily of HIT proteins, which are characterized by an histidine-triad sequence motif. While the eukaryotic variants hydrolyze AMP derivates and modulate transcription, the function of prokaryotic HinT proteins is less clearly defined. In Mycoplasma hominis, HinT is concomitantly expressed with the proteins P60 and P80, two domains of a surface exposed membrane complex, and in addition interacts with the P80 moiety. RESULTS: An cluster of hitABL genes, similar to that of M. hominis was found in M. pulmonis, M. mycoides subspecies mycoides SC, M. mobile and Mesoplasma florum. RT-PCR analyses provided evidence that the P80, P60 and HinT homologues of M. pulmonis were polycistronically organized, suggesting a genetic and physical interaction between the proteins encoded by these genes in these species. While the hit loci of M. pneumoniae and M. genitalium encoded, in addition to HinT, a protein with several transmembrane segments, the hit locus of Ureaplasma parvum encoded a pore-forming protein, UU270, a P60 homologue, UU271, HinT, UU272, and a membrane protein of unknown function, UU273. Although a full-length mRNA spanning the four genes was not detected, amplification of all intergenic regions from the center of UU270 to the end of UU273 by RT-PCR may be indicative of a common, but unstable mRNA. In Chlamydiaceae the hit gene is flanked upstream by a gene predicted to encode a metal dependent hydrolase and downstream by a gene putatively encoding a protein with ARM-repeats, which are known to be involved in protein-protein interactions. In RT-PCR analyses of C. pneumoniae, regions comprising only two genes, Cp265/Cp266 and Cp266/Cp267 were able to be amplified. In contrast to this in vivo interaction analysis using the yeast two-hybrid system and in vitro immune co-precipitation revealed an interaction between Cp267, which contains the ARM repeats, Cp265, the predicted hydrolase, and Cp266, the HinT protein. CONCLUSION: In the Mollicutes HinT proteins were shown to be linked with membrane proteins while in the Chlamydiaceae they were genetically and physically associated with cytoplasmic proteins, one of which is predicted to be a metal-dependent phosphoesterase. Future work will elucidate whether these differing associations indicate that HinT proteins have evolved independently or are indeed two hotspots of a common sphere of action of bacterial HinT proteins. PMID- 15904497 TI - Fungal-specific PCR primers developed for analysis of the ITS region of environmental DNA extracts. AB - BACKGROUND: The Internal Transcribed Spacer (ITS) regions of fungal ribosomal DNA (rDNA) are highly variable sequences of great importance in distinguishing fungal species by PCR analysis. Previously published PCR primers available for amplifying these sequences from environmental samples provide varying degrees of success at discriminating against plant DNA while maintaining a broad range of compatibility. Typically, it has been necessary to use multiple primer sets to accommodate the range of fungi under study, potentially creating artificial distinctions for fungal sequences that amplify with more than one primer set. RESULTS: Numerous sequences for PCR primers were tested to develop PCR assays with a wide range of fungal compatibility and high discrimination from plant DNA. A nested set of 4 primers was developed that reflected these criteria and performed well amplifying ITS regions of fungal rDNA. Primers in the 5.8S sequence were also developed that would permit separate amplifications of ITS1 and ITS2. A range of basidiomycete fruiting bodies and ascomycete cultures were analyzed with the nested set of primers and Restriction Fragment Length Polymorphism (RFLP) fingerprinting to demonstrate the specificity of the assay. Single ectomycorrhizal root tips were similarly analyzed. These primers have also been successfully applied to Quantitative PCR (QPCR), Length Heterogeneity PCR (LH-PCR) and Terminal Restriction Fragment Length Polymorphism (T-RFLP) analyses of fungi. A set of wide-range plant-specific primers were developed at positions corresponding to one pair of the fungal primers. These were used to verify that the host plant DNA was not being amplified with the fungal primers. CONCLUSION: These plant primers have been successfully applied to PCR-RFLP analyses of forest plant tissues from above- and below-ground samples and work well at distinguishing a selection of plants to the species level. The complete set of primers was developed with an emphasis on discrimination between plant and fungal sequences and should be particularly useful for studies of fungi where samples also contain high levels of background plant DNA, such as verifying ectomycorrhizal morphotypes or characterizing phylosphere communities. PMID- 15904498 TI - Synergistic effect of intrathecal fentanyl and bupivacaine in spinal anesthesia for cesarean section. AB - BACKGROUND: Potentiating the effect of intrathecal local anesthetics by addition of intrathecal opiods for intra-abdominal surgeries is known. In this study by addition of fentanyl we tried to minimize the dose of bupivacaine, thereby reducing the side effects caused by higher doses of intrathecal bupivacaine in cesarean section. METHODS: Study was performed on 120 cesarean section parturients divided into six groups, identified as B8, B10 and B 12.5 8.10 and 12.5 mg of bupivacaine mg and FB8, FB10 and FB 12.5 received a combination of 12.5 mug intrathecal fentanyl respectively. The parameters taken into consideration were visceral pain, hemodynamic stability, intraoperative sedation, intraoperative and postoperative shivering, and postoperative pain. RESULTS: Onset of sensory block to T6 occurred faster with increasing bupivacaine doses in bupivacaine only groups and bupivacaine -fentanyl combination groups. Alone lower concentrations of bupivacaine could not complete removed the visceral pain. Blood pressure declined with the increasing concentration of Bupivacaine and Fentanyl. Incidence of nausea and shivering reduces significantly whereas, the postoperative pain relief and hemodynamics increased by adding fentanyl. Pruritis, maternal respiratory depression and changes in Apgar score of babies do not occur with fentanyl. CONCLUSION: Spinal anesthesia among the neuraxial blocks in obstetric patients needs strict dose calculations because minimal dose changes, complications and side effects arise, providing impetus for this study. Here the synergistic, potentiating effect of fentanyl (an opiod) on bupivacaine (a local anesthetic) in spinal anesthesia for cesarian section is presented, fentanyl is able to reduce the dose of bupivacaine and therefore its harmful effects. PMID- 15904499 TI - Recruitment of ethnic minority patients to a cardiac rehabilitation trial: the Birmingham Rehabilitation Uptake Maximisation (BRUM) study [ISRCTN72884263]. AB - BACKGROUND: Concerns have been raised about low participation rates of people from minority ethnic groups in clinical trials. However, the evidence is unclear as many studies do not report the ethnicity of participants and there is insufficient information about the reasons for ineligibility by ethnic group. Where there are data, there remains the key question as to whether ethnic minorities more likely to be ineligible (e.g. due to language) or decline to participate. We have addressed these questions in relation to the Birmingham Rehabilitation Uptake Maximisation (BRUM) study, a randomized controlled trial (RCT) comparing a home-based with a hospital-based cardiac rehabilitation programme in a multi-ethnic population in the UK. METHODS: Analysis of the ethnicity, age and sex of presenting and recruited subjects for a trial of cardiac rehabilitation in the West-Midlands, UK. PARTICIPANTS: 1997 patients presenting post-myocardial infarction, percutaneous transluminal coronary angioplasty or coronary artery bypass graft surgery. DATA COLLECTED: Exclusion rates, reasons for exclusion and reasons for declining to participate in the trial by ethnic group. RESULTS: Significantly more patients of South Asian ethnicity were excluded (52% of 'South Asian' v 36% 'White European' and 36% 'Other', p < 0.001). This difference in eligibility was primarily due to exclusion on the basis of language (i.e. the inability to speak English or Punjabi). Of those eligible, similar proportions were recruited from the different ethnic groups (white, South Asian and other). There was a marked difference in eligibility between people of Indian, Pakistani or Bangladeshi origin. CONCLUSION: Once eligible for this trial, people from different ethnic groups were recruited in similar proportions. The reason for ineligibility in the BRUM study was the inability to support the range of minority languages. PMID- 15904500 TI - General practitioners' conceptions about treatment of depression and factors that may influence their practice in this area. A postal survey. AB - BACKGROUND: The way GPs work does not appear to be adapted to the needs of depressive patients. Therefore we wanted to examine Swedish GPs' conceptions of depressive disorders and their treatment and GPs' ideas of factors that may influence their manner of work with depressive patients. METHODS: A postal questionnaire to a stratified sample of 617 Swedish GPs. RESULTS: Most respondents assumed antidepressive drugs effective and did not assume that psychotherapy can replace drugs in depression treatment though many of them looked at psychotherapy as an essential complement. Nearly all respondents thought that clinical experiences had great importance in decision situations, but patients' own preferences and official clinical guidelines were also regarded as essential. As influences on their work, almost all surveyed GPs regarded experiences from general practice very important, and a majority also emphasised experiences from private life. Courses arranged by pharmaceutical companies were seen as essential sources of knowledge. A majority thought that psychiatrists did not provide sufficient help, while most respondents perceived they were well backed up by colleagues. CONCLUSION: GPs tend to emphasize experiences, both from clinical work and private life, and overlook influences of collegial dealings and ongoing CME as well as the effects of the pharmaceutical companies' marketing activities. Many GPs appear to need more evidence based knowledge about depressive disorders. Interventions to improve depression management have to be supporting and interactive, and should be combined with organisational reforms to improve co-operation with psychiatrists. PMID- 15904501 TI - Prevalence and risk factors of syphilis infection among drug addicts. AB - BACKGROUND: Recent epidemiological data show an increased trend of official estimates for syphilis infection in the general population. Many of the infected cases remain undetected leaving an underestimation of the true prevalence of syphilis in the general population, but also among subpopulations such as illicit drug users. There is limited epidemiological data published on the proportion and risk factors of syphilis infections associated with illicit drug abuse. METHODS: Illicit drug addicts (n = 1223) in inpatients units in Germany were screened (2000-01) for syphilis and interviewed regarding patterns of drug use and sexual behaviour. TPHA-test for initial screening and FTA-ABS-IgM test in TPHA-positive patients were used. RESULTS: In total, TPHA-tests were positive in 39 (3.3%) and 7 patients (0.6%) were IgM positive. The prevalence rate for syphilis in males was 1.9% and for women it was 8.5%. Female patients were 4.56 (CI 95% 2.37-8.78) times more likely to have a positive TPHA test than males. Sexual behaviours such as high number of sexual partners, sex for drugs/money, sex on the first day were associated with syphilis infection only in women. Females with frequent sex for drugs or money had 4.31 (CI 95% 2.32-8.52) times more likely a reactive TPHA test than remaining patients. Neither the sociodemographic factors nor sexual behaviour were statistically significant associated with syphilis infection among men at all. CONCLUSION: Our data suggest the need for screening for syphilis among these illicit drug users in inpatient settings, in particular among sexual active women. This conclusion is corroborated by the finding of increasing numbers of syphilis infections in the general population. The identification of syphilis cases among drug addicts would give treatment options to these individuals and would help to reduce the spread of infection in this population, but also a spread into heterosexual populations related to prostitution. PMID- 15904502 TI - Echinococcal disease in Alberta, Canada: more than a calcified opacity. AB - BACKGROUND: Most cases of echinococcal disease (ED) acquired in Canada are thought to be due to the sylvatic form of Echinococcus granulosus, which may be more benign than ED due to either Echinococcus multilocularis or the pastoral form of E. granulosus. There are limited descriptions of the clinical course and outcome of Canadian patients with ED in the modern era. METHODS: A retrospective chart review was performed of patients hospitalized with echinococcal disease (ED) from 1991 to 2001 in Edmonton, Alberta. RESULTS: Forty-two cases of ED were identified of which 19 were definite, 3 probable, and 20 possible. Further analysis was limited to the 22 definite and probable cases, of which 77% were female and 41% aboriginal, with an age range of 5 to 87 years. Nine patients (40%) had pulmonary involvement and 11 (50%) hepatic involvement. One patient had an intracardiac mass presenting as a cerebrovascular event and one had a splenic cyst. Seven of the 22 patients had combined surgical resection and medical treatment, six had surgical resection of the cyst alone, four had cyst aspiration, one had medical treatment alone and four had no specific treatment. There was no mortality attributable to ED but three patients died of unrelated illnesses. CONCLUSION: Echinococcal disease in northern Alberta has a marked diversity of clinical presentations, and generally has a good prognosis despite a wide variety of therapeutic interventions. PMID- 15904503 TI - Blood cultures in ambulatory outpatients. AB - BACKGROUND: Blood cultures are a gold standard specific test for diagnosing many infections. However, the low yield may limit their usefulness, particularly in low-risk populations. This study was conducted to assess the utility of blood cultures drawn from ambulatory outpatients. METHODS: Blood cultures drawn at community-based collection sites in the Calgary Health Region (population 1 million) in 2001 and 2002 were included in this study. These patients were analyzed by linkages to acute care health care databases for utilization of acute care facilities within 2 weeks of blood culture draw. RESULTS: 3102 sets of cultures were drawn from 1732 ambulatory outpatients (annual rate = 89.4 per 100,000 population). Significant isolates were identified from 73 (2.4%) sets of cultures from 51 patients, including Escherichia coli in 18 (35%) and seven (14%) each of Staphylococcus aureus and Streptococcus pneumoniae. Compared to patients with negative cultures, those with positive cultures were older (mean 49.6 vs. 40.1 years, p < 0.01), and more likely to subsequently receive care at a regional emergency department, outpatient antibiotic clinic, or hospital (35/51 vs. 296/1681, p < 0.0001). Of the 331 (19%) patients who received acute care treatment, those with positive cultures presented sooner after community culture draw (median 2 vs. 3 days, p < 0.01) and had longer median treatment duration (6 vs. 2 days, p < 0.01). CONCLUSION: Blood cultures drawn in outpatient settings are uncommonly positive, but may define patients for increased intensity of therapy. Strategies to reduce utilization without excluding patients with positive cultures need to be developed for this patient population. PMID- 15904504 TI - Hepatitis E virus infection in hemodialysis patients: a seroepidemiological survey in Iran. AB - BACKGROUND: The hepatitis E virus (HEV) has a global distribution and is known to have caused large waterborne epidemics of icteric hepatitis. Transmission is generally via the fecal-oral route. Some reports have suggested parenteral transmission of HEV. Anti-HEV prevalence data among chronic hemodialysis (HD) patients are few and give conflicting results. METHODS: This cross-sectional study was conducted in August of 2004. We tested 324 chronic HD patients attending three different units in the city of Tabriz, northwestern part of Iran, for anti-HEV antibody. A specific solid- phase enzyme-linked immunoassay (Diapro, Italy) was used. RESULTS: The overall seroprevalence of hepatitis E was 7.4 %(95% CI: 4.6%-10.6%). The prevalence rate of HBV and HCV infection were 4.6% (95% CI: 2.3%-6.9%) and 20.4% (95% CI: 16%-24.8%), respectively. No significant association was found between anti-HEV positivity and age, sex, duration of hemodialysis, positivity for hepatitis B or C virus infection markers and history of transfusion. CONCLUSION: We observed high anti-HEV antibody prevalence; there was no association between HEV and blood borne infections (HBV, HCV, and HIV) in our HD patients. This is the first report concerning seroepidemiology of HEV infection in a large group of chronic HD individuals in Iran. PMID- 15904505 TI - Clinical outcomes in typhoid fever: adverse impact of infection with nalidixic acid-resistant Salmonella typhi. AB - BACKGROUND: Widespread use of fluoroquinolones has resulted in emergence of Salmonella typhi strains with decreased susceptibility to fluoroquinolones. These strains are identifiable by their nalidixic acid-resistance. We studied the impact of infection with nalidixic acid-resistant S. typhi (NARST) on clinical outcomes in patients with bacteriologically-confirmed typhoid fever. METHODS: Clinical and laboratory features, fever clearance time and complications were prospectively studied in patients with blood culture-proven typhoid fever, treated at a tertiary care hospital in north India, during the period from November 2001 to October 2003. Susceptibility to amoxycillin, co-trimoxazole, chloramphenicol, ciprofloxacin and ceftriaxone were tested by disc diffusion method. Minimum inhibitory concentrations (MIC) of ciprofloxacin and ceftriaxone were determined by E-test method. RESULTS: During a two-year period, 60 patients (age [mean +/- SD]: 15 +/- 9 years; males: 40 [67%]) were studied. All isolates were sensitive to ciprofloxacin and ceftriaxone by disc diffusion and MIC breakpoints. However, 11 patients had clinical failure of fluoroquinolone therapy. Infections with NARST isolates (47 [78%]) were significantly associated with longer duration of fever at presentation (median [IQR] 10 [7-15] vs. 4 [3-6] days; P = 0.000), higher frequency of hepatomegaly (57% vs. 15%; P = 0.021), higher levels of aspartate aminotransferase (121 [66-235] vs. 73 [44-119] IU/L; P = 0.033), and increased MIC of ciprofloxacin (0.37 +/- 0.21 vs. 0.17 +/- 0.14 microg/mL; P = 0.005), as compared to infections with nalidixic acid-susceptible isolates. All 11 patients with complications were infected with NARST isolates. Total duration of illness was significantly longer in patients who developed complications than in patients who did not (22 [14.8-32] vs. 12 [9.3-20.3] days; P = 0.011). Duration of prior antibiotic intake had a strong positive correlation with the duration of fever at presentation (r = 0.61; P = 0.000) as well as the total duration of illness (r = 0.53; P = 0.000). CONCLUSION: Typhoid fever caused by NARST infection is associated with poor clinical outcomes, probably due to delay in initiating appropriate antibiotic therapy. Fluoroquinolone breakpoints for S. typhi need to be redefined and fluoroquinolones should no longer be used as first-line therapy, if the prevalence of NARST is high. PMID- 15904506 TI - Subtelomeric study of 132 patients with mental retardation reveals 9 chromosomal anomalies and contributes to the delineation of submicroscopic deletions of 1pter, 2qter, 4pter, 5qter and 9qter. AB - BACKGROUND: Cryptic chromosome imbalances are increasingly acknowledged as a cause for mental retardation and learning disability. New phenotypes associated with specific rearrangements are also being recognized. Techniques for screening for subtelomeric rearrangements are commercially available, allowing the implementation in a diagnostic service laboratory. We report the diagnostic yield in a series of 132 subjects with mental retardation, and the associated clinical phenotypes. METHODS: We applied commercially available subtelomeric fluorescence in situ hybridization (FISH). All patients referred for subtelomeric screening in a 5-year period were reviewed and abnormal cases were further characterized clinically and if possible molecularly. RESULTS: We identified nine chromosomal rearrangements (two of which were in sisters) corresponding to a diagnostic yield of approx. 7%. All had dysmorphic features. Five had imbalances leading to recognizable phenotypes. CONCLUSION: Subtelomeric screening is a useful adjunct to conventional cytogenetic analyses, and should be considered in mentally retarded subjects with dysmorphic features and unknown cause. PMID- 15904507 TI - Reevaluation of the 22-1-1 antibody and its putative antigen, EBAG9/RCAS1, as a tumor marker. AB - BACKGROUND: Tumor-associated antigens are appreciated as diagnostic markers, but they have also prompted tremendous efforts to develop tumor-specific immunotherapy. A previously cloned tumor-associated antigen, EBAG9, was initially defined by reactivity with the monoclonal antibody 22-1-1. Functionally, the EBAG9-encoded gene-product was believed to induce apoptosis in activated immune cells. However, using a cell-biological approach we identified EBAG9 as a Golgi resident modulator of O-linked glycan expression, the latter product was then recognized by the 22-1-1 antibody. Secondly, EBAG9 expression was found physiologically in all murine tissues examined. This raised the question if EBAG9 is tumor-specific and mediates apoptosis itself or through O-linked glycans generated, among them the cognate 22-1-1 antigen Tn. METHODS: We have used immunohistochemistry to detect the expression of 22-1-1 and EBAG9 in various tissues. Correlation between expression of both antigens in cell lines was analysed by immunoblot and flow cytometry. Apoptosis was studied by using flow cytometry and Caspase-Glo 3/7 assay kit. Cellular distribution of EBAG9 was analysed by electron and confocal microscopy. RESULTS: Here, we compared expression of the 22-1-1 and EBAG9-defined antigens in normal and neoplastic tissues in situ. In contrast to 22-1-1 staining, EBAG9 is a ubiquitously expressed antigen in all normal and cancerous tissues. Functional studies on the role of 22-1-1 reactive material did not support any evidence for apoptosis induction. Employing electron and confocal microscopy, a refined subcellular localization of EBAG9 at the Golgi was obtained. CONCLUSION: We suggest that the estrogen-inducible EBAG9 gene-product and the 22-1-1 defined antigen are structurally and functionally separate antigens. PMID- 15904508 TI - Minimum follow-up time required for the estimation of statistical cure of cancer patients: verification using data from 42 cancer sites in the SEER database. AB - BACKGROUND: The present commonly used five-year survival rates are not adequate to represent the statistical cure. In the present study, we established the minimum number of years required for follow-up to estimate statistical cure rate, by using a lognormal distribution of the survival time of those who died of their cancer. We introduced the term, threshold year, the follow-up time for patients dying from the specific cancer covers most of the survival data, leaving less than 2.25% uncovered. This is close enough to cure from that specific cancer. METHODS: Data from the Surveillance, Epidemiology and End Results (SEER) database were tested if the survival times of cancer patients who died of their disease followed the lognormal distribution using a minimum chi-square method. Patients diagnosed from 1973-1992 in the registries of Connecticut and Detroit were chosen so that a maximum of 27 years was allowed for follow-up to 1999. A total of 49 specific organ sites were tested. The parameters of those lognormal distributions were found for each cancer site. The cancer-specific survival rates at the threshold years were compared with the longest available Kaplan-Meier survival estimates. RESULTS: The characteristics of the cancer-specific survival times of cancer patients who died of their disease from 42 cancer sites out of 49 sites were verified to follow different lognormal distributions. The threshold years validated for statistical cure varied for different cancer sites, from 2.6 years for pancreas cancer to 25.2 years for cancer of salivary gland. At the threshold year, the statistical cure rates estimated for 40 cancer sites were found to match the actuarial long-term survival rates estimated by the Kaplan-Meier method within six percentage points. For two cancer sites: breast and thyroid, the threshold years were so long that the cancer-specific survival rates could yet not be obtained because the SEER data do not provide sufficiently long follow-up. CONCLUSION: The present study suggests a certain threshold year is required to wait before the statistical cure rate can be estimated for each cancer site. For some cancers, such as breast and thyroid, the 5- or 10-year survival rates inadequately reflect statistical cure rates, and highlight the need for long-term follow-up of these patients. PMID- 15904509 TI - Female breast cancer incidence and survival in Utah according to religious preference, 1985-1999. AB - BACKGROUND: Female breast cancer incidence rates in Utah are among the lowest in the U.S. The influence of the Church of Jesus Christ of Latter-day Saint (LDS or Mormon) religion on these rates, as well as on disease-specific survival, will be explored for individuals diagnosed with breast cancer in Utah from 1985 through 1999. METHODS: Population-based records for incident female breast cancer patients were linked with membership records from the LDS Church to determine religious affiliation and, for LDS Church members, level of religiosity. Incidence rates were age-adjusted to the 2000 U.S. standard population using the direct method. Cox proportional hazards model was used to compare survival among religiously active LDS, less religiously active LDS, and non-LDS with simultaneous adjustment for prognostic factors. RESULTS: Age-adjusted breast cancer incidence rates were consistently lower for LDS than non-LDS in Utah from 1985 through 1999. Rates were lower among LDS compared with non-LDS across the age span. In 1995-99, the age-adjusted incidence rates were 107.6 (95% CI: 103.9 111.3) for LDS women and 130.5 (123.2 - 137.9) for non-LDS women. If non-LDS women in Utah had the same breast cancer risk profile as LDS women, an estimated 214 (4.8%) fewer malignant breast cancer cases would have occurred during 1995 99. With religiously active LDS serving as the reference group, the adjusted death hazard ratio for religiously less active LDS was 1.09 (0.94 - 1.27) and for non-LDS was 0.86 (0.75 - 0.98). CONCLUSION: In Utah, LDS lifestyle is associated with lower incidence rates of female breast cancer. However, LDS experience poorer survivability from breast cancer than their non-LDS counterparts. Parity and breastfeeding, while protective factors against breast cancer, may contribute to poorer prognosis of female breast cancer in LDS women. PMID- 15904510 TI - Early discontinuation of intravenous antimicrobial therapy in pediatric oncology patients with febrile neutropenia. AB - BACKGROUND: There are no standard criteria for when to discontinue intravenous antimicrobial therapy (IVAMT) in children with febrile neutropenia (FN), but it is now common to discontinue IVAMT and discharge patients with an absolute neutrophil count (ANC) < or = 500/mm3. The purpose of this study was to evaluate the outcome of a large cohort of children with FN who had IVAMT discontinued with an ANC < or = 500/mm3 METHODS: A retrospective chart review was completed of patients in the Northern Alberta Children's Cancer Program with FN and no apparent clinical source of fever from June 1, 1997 to July 1, 2002. RESULTS: Out of a total of 275 patients, 127 (46%) had at least one episode of FN, with FN occurring in patients with sarcomas more commonly than in those with leukemia/ lymphoma and least in those with other solid tumors. In 59 of 276 episodes of FN (21%) patients had a microbiologically defined infection at admission. Of the 217 remaining episodes, 112 of 199 patients (56%) with known neutrophil counts had IVAMT discontinued before their absolute neutrophil count (ANC) reached 500/mm3 at the discretion of the clinician. Fever recurred in only two of these patients after discharge, and there were no bacterial infections diagnosed after parenteral antibiotics were discontinued. CONCLUSION: Even without use of standard criteria for early discharge, clinicians appear to be skilled at selecting children with FN who can safely have IVAMT discontinued with an ANC < or = 500/mm3. PMID- 15904511 TI - Rural residence is not a risk factor for frequent mental distress: a behavioral risk factor surveillance survey. AB - BACKGROUND: Residents of rural areas may be at increased risk of mental health problems. If so, public health programs aimed at preventing poor mental health may have to be customized for delivery to rural areas. The purpose of this study was to examine the relationship between residing in a rural area and frequent mental distress, which is one indicator of poor mental health. METHODS: The Behavioral Risk Factor Surveillance System (BRFSS) survey for the state of Texas was the source of information about obesity, demographic characteristics, and frequent mental distress (FMD). FMD was defined as poor self-rated mental health during at least half of the days in the last month. Adjusted odds for FMD were computed for rural and suburban respondents relative to urban respondents. RESULTS: FMD was found to be independently associated with lower education, being younger, being non-Hispanic, being unmarried, and being female. FMD also was associated with being obese or underweight and suburban residence (relative to metro-central city). FMD was not more common among rural respondents than in the metro-central city. CONCLUSION: Rural respondents were not at greater risk of frequent mental distress than urban respondents in this sample. Programs seeking to improve community mental health should target persons with less education and extremes in body weight, along with women and single persons, regardless of whether they live in rural or urban areas. PMID- 15904512 TI - Prevalence of nonvitamin, nonmineral supplement usage among students in a Turkish university. AB - BACKGROUND: There have been multiple studies carried out in many countries with regard to the use of nonvitamin, nonmineral (NVNM) supplements. These studies have shown that the use of NVNM supplements is on the increase throughout the world, particularly in western countries. The aim of this study was to assess the extent of NVNM supplement use among Turkish university students. METHODS: The survey was conducted between September and December 2004 at Osmangazi University, a public university located in the west of Turkey. Responses were analysed, using the chi-square (x2) test, t test and percent (%) ratios, according to gender and consumers. Differences were considered significant for p